PMID- 11882901 TI - Ubiquitination-dependent cofactor exchange on LIM homeodomain transcription factors. AB - The interactions of distinct cofactor complexes with transcription factors are decisive determinants for the regulation of gene expression. Depending on the bound cofactor, transcription factors can have either repressing or transactivating activities. To allow a switch between these different states, regulated cofactor exchange has been proposed; however, little is known about the molecular mechanisms that are involved in this process. LIM homeodomain (LIM-HD) transcription factors associate with RLIM (RING finger LIM domain-binding protein) and with CLIM (cofactor of LIM-HD proteins; also known as NLI, Ldb and Chip) cofactors. The co-repressor RLIM inhibits the function of LIM-HD transcription factors, whereas interaction with CLIM proteins is important for the exertion of the biological activity conferred by LIM-HD transcription factors. Here we identify RLIM as a ubiquitin protein ligase that is able to target CLIM cofactors for degradation through the 26S proteasome pathway. Furthermore, we demonstrate a ubiquitination-dependent association of RLIM with LIM-HD proteins in the presence of CLIM cofactors. Our data provide a mechanistic basis for cofactor exchange on DNA-bound transcription factors, and probably represent a general mechanism of transcriptional regulation. PMID- 11882902 TI - Structure of the HP1 chromodomain bound to histone H3 methylated at lysine 9. AB - Specific modifications to histones are essential epigenetic markers---heritable changes in gene expression that do not affect the DNA sequence. Methylation of lysine 9 in histone H3 is recognized by heterochromatin protein 1 (HP1), which directs the binding of other proteins to control chromatin structure and gene expression. Here we show that HP1 uses an induced-fit mechanism for recognition of this modification, as revealed by the structure of its chromodomain bound to a histone H3 peptide dimethylated at Nzeta of lysine 9. The binding pocket for the N-methyl groups is provided by three aromatic side chains, Tyr21, Trp42 and Phe45, which reside in two regions that become ordered on binding of the peptide. The side chain of Lys9 is almost fully extended and surrounded by residues that are conserved in many other chromodomains. The QTAR peptide sequence preceding Lys9 makes most of the additional interactions with the chromodomain, with HP1 residues Val23, Leu40, Trp42, Leu58 and Cys60 appearing to be a major determinant of specificity by binding the key buried Ala7. These findings predict which other chromodomains will bind methylated proteins and suggest a motif that they recognize. PMID- 11882903 TI - The role of boost irradiation in the conservative treatment of stage I-II breast cancer. AB - In this article, we review the current status, indication, technical aspects, controversies, and future prospects of boost irradiation after breast conserving surgery (BCS). BCS and radiotherapy (RT) of the conserved breast became widely accepted in the last decades for the treatment of early invasive breast cancer. The standard technique of RT after breast conservation is to treat the whole breast up to a total dose of 45 to 50 Gy. However, there is no consensus among radiation oncologists about the necessity of boost dose to the tumor bed. Generally accepted criteria for identification of high risk subgroups, in which boost is recommended, have not been established yet. Further controversy exists regarding the optimal boost technique (electron vs. brachytherapy), and their impact on local tumor control and cosmesis. Based on the results of numerous retrospective and recently published prospective trials, the European brachytherapy society (GEC-ESTRO), as well as the American Brachytherapy Society has issued their guidelines in these topics. These guidelines will help clinicians in their medical decisions. Some aspects of boost irradiation still remain somewhat controversial. The final results of prospective boost trials with longer follow-up, involving analyses based on pathologically defined subgroups, will clarify these controversies. Preliminary results with recently developed boost techniques (intraoperative RT, CT-image based 3D conformal brachytherapy, and 3D virtual brachytherapy) are promising. However, more experience and longer follow-up are required to define whether these methods might improve local tumor control for breast cancer patients treated with conservative surgery and RT. PMID- 11882904 TI - Expression of thrombospondin-1 in human pancreatic adenocarcinomas: role in matrix metalloproteinase-9 production. AB - Human pancreatic adenocarcinoma, an aggressive malignant disease, shows a strong desmoplastic reaction characterized by a remarkable proliferation of interstitial connective tissues. Thrombospondin-1 (TSP-1), a 450 kDa platelet and matrix glycoprotein, has been implicated in tumor invasion, angiogenesis and metastasis. TSP-1 and MMP-9 expression in pancreatic adenocarcinoma and control pancreas tissues was measured by immunohistochemistry. TSP-1 expression in pancreatic carcinoma cell lines, fibroblasts, and endothelial cells was measured by a competitive TSP-1 enzyme linked immunosorbent assay (ELISA). The effect of TSP-1 on MMP-9 production in pancreatic carcinoma cell lines was measured by zymography and Western blot analysis. Eighty five per cent (23/27) of cases of pancreatic adenocarcinoma showed increased TSP-1 staining in the desmoplastic stroma adjacent to tumor cells. No specific positive staining for TSP-1 was observed in the normal pancreatic tissues and the inflammatory areas. TSP-1 localized in tumor stroma surrounding the tumor cells expressing MMP-9. Using TSP-1 competitive ELISA, the secretion of TSP-1 by different pancreatic cancer cell lines into culture medium varied from 11.45 plus minus 14.08 to 275.82 plus minus 45.56 ng/10 6 cells/24 hours. The amounts of TSP-1 detected in both culture media and cell extracts from fibroblasts or endothelial cells were at least 2-3 fold higher than those from pancreatic cancer cells. TSP-1 augmented the production of matrix metalloproteinase-9, a matrix degrading enzyme, in pancreatic cancer cells in vitro. Stromally-derived TSP-1 up-regulates the production of MMP-9 by pancreatic adenocarcinoma. These data are consistent with the conclusion that TSP 1-rich stroma is involved in regulating matrix remodeling in tumor invasion. PMID- 11882905 TI - Expression of a decorin-like moleculein human melanoma. AB - Decorin, a member of the family of small leucin-rich proteoglycans, has originally been described as a secreted proteoglycan component of the connective tissues, and has been implicated in the negative regulation of cell proliferation directly or via interactions with TGF-beta. It was reported to be generally absent from tumor cells. Here we show that human melanoma cell lines express a decorin-like molecule. We detected decorin mRNA by RT-PCR in 7 out 7 human melanoma lines characterized by various metastatic potential. Using polyclonal antiserum against the core protein of decorin, the typical 80-120 kD glycanated form as well as a high molecular weight aberrant version (200-210 kD) of decorin were demonstrated by Western blot technique in the culture supernatants as well as in lysates of human melanoma cells. Finally, decorin epitope was also demonstrated immunohistochemically in human melanoma xenografts, as well as in tumor cells of surgically resected melanomas but not in melanocytes of nevi. The expression of this aberrant decorin did not inhibit the in vitroor in vivogrowth of human melanoma cells, and it was independent of their metastatic potential. Human melanoma cell lines expressing aberrant decorin retained sensitivity to the antiproliferative and gelatinase-stimulatory effects of exogenous TGF-beta. PMID- 11882906 TI - Clinicopathological features, MIB-1 labeling index and apoptotic index in recurrent astrocytic tumors. AB - This is a study of 64 cases of recurrent astrocytic tumors of all four WHO grades wherein a comparative evaluation of initial vs. recurrent tumor was done with respect to histological grading, MIB-1 labeling index (LI) and apoptotic index (AI). The aim was to identify factor/s that could influence interval to recurrence and/or malignant progression. Recurrence was noted in all grades and upon recurrence, 93.3% of grade II (low grade diffuse) astrocytomas and 63.6% of grade III anaplastic astrocytomas underwent malignant progression. However, none of the Grade I tumors showed evidence of malignant progression. Though interval to recurrence varied considerably, there was a correlation with histological grade of the initial tumor in that grade I and II tumors had a significantly longer mean interval to recurrence (43 months and 54.8 months respectively) as compared to grade III and IV (glioblastoma multiforme) tumors (17.6 and 12.8 months respectively). The interval to recurrence was also longer for grade II and III tumors which showed progression on recurrence (55.3 months for Grade II >Grade III; 54 months for Grade II->Grade IV and 20.6 months for Grade III->IV) as compared to tumors which recurred to the same grade (12.5 months for Grade III >Grade III and 12.8 months for Grade IV->Grade IV). A statistically significant inverse correlation of MIB-1 LI with interval to recurrence was noted. Higher the MIB-1 LI, shorter was the interval to recurrence. Further a cut off MIB-1 LI value of 2.8% could be proposed in predicting recurrence free survival. Interestingly, MIB-1 LI of grade II tumors, which had progressed to grade IV was significantly higher than MIB-1 LI of grade II tumors which had progressed to grade III. Thus, this study establishes the potential role of MIB-1 LI of the initial tumor in determining interval to recurrence. However, apoptotic index has no role in predicting either interval to recurrence or malignant progression. PMID- 11882907 TI - Her-2/neu gene amplification compared with HER-2/neu protein overexpression on ultrasound guided core-needle biopsy specimens of breast carcinoma. AB - Genomic amplification and oncoprotein overexpression of Her-2/neu was studied on ultrasound core needle biopsy specimens of the infiltrative ductal carcinomas of the breast. We performed two colour fluorescence in situ hybridization (FISH) for Her-2/neu and chromosome 17 and compared the FISH results with the immunohistochemical overexpression of Her-2/neu protein by 2 antibodies (DAKO HercepTest and the BioGenex monoclonal antibody AM 134-5M). Furthermore, following radical mastectomy with axillary dissection, Her-2/neu status of the patients were compared with the well known histopathological prognostic factors such as histologic grade, tumor stage, lympho/ vascular invasion, surgical margin status and Paget s disease. Amplification was demonstrated 27% of the cases. Her 2/neu protein overexpression was detected in 47% and 80% of the cases with CB11 and HercepTest respectively. We revealed statistically significant association between the tumor, oncoprotein expression and oncogene amplification (p<0.05). The results of our study showed that combination of IHC and FISH methods enhances the evaluation of tumor genetics at both gene and protein level for the analysis of Her-2/neu in breast carcinoma. PMID- 11882908 TI - MUC1 mucin and carbohydrate associated antigens as tumor markers in head and neck squamous cell carcinoma. AB - An immunological analysis to study MUC1 mucin core protein and carbohydrate associated antigens as tissue tumor markers in head and neck carcinoma was performed. Twenty nine patients with the following tumor localizations were included: tongue (n=10), larynx (n=8), oral cavity (n=4), maxillary sinus (n=3), tonsillar ring (n=3) and pharynx (n=1); seven samples of epithelium obtained from normal organs at the same localizations were studied as controls. Immunohistochemical analysis was performed following standard procedures and reaction was graded according to staining intensity and distribution. From each tissue section, membrane, cytoplasmic and nuclear moieties were obtained by differential centrifugation with subsequent fractionation by density gradient centrifugation (6M guanidium chloride-CsCl); subcellular moieties and CsCl derived fractions were analyzed by immunoblotting. Monoclonal antibodies (MAbs) reacting with the core protein of MUCI (C595) and associated carbohydrate antigens were: Tn, 83D4 MAb; Lewis y antigen (Le y), C14 MAb; Lewis x antigen (Le x), KM380 MAb and sialyl Lewis x (sLe x), KM93 MAb. Statistical analysis was undertaken by Spearman rank correlation. In tumor samples, the immunohistochemical identification of MUCl core protein and associated antigens was extended; differences were found in the pattern and intensity of expression; results were corroborated by immunoblotting although in a few samples there was not coincidence between both methods. Localization, tumor mass or node involvement did not show significant differences for any of the antigens studied. CONCLUSIONS: 1) head and neck carcinoma expressed MUCI and associated carbohydrate antigens in high levels; 2) no relationship between antigenic expression and tumor status was found. PMID- 11882910 TI - Limitations of fine-needle aspiration cytology to diagnose metaplastic carcinoma of the breast. AB - Metaplastic carcinoma is a very rare breast neoplasm that is often confused with benign and others malignant entities on both clinical and conventional histopathologic basis. Three cases of metaplastic carcinoma of breast are presented. The difficulties found on fine needle aspiration cytology and the limitations of this procedure are discussed as well the main features of this tumor. PMID- 11882909 TI - New diagnostic tool for differentiation of idiopathic hypereosinophilic syndrome (HES) and secondary eosinophilic states. AB - The hypereosinophilic syndrome (HES) is a very rare disease, characterized by persistent eosinophilia with tissue involvement and organ dysfunction which often precedes a subsequent T cell lymphoma. Interleukin-5 secreted by a T lymphocyte subpopulation has been described in previous reports as the most important factor responsible for the prolonged lifespan of the eosinophils. The goal of the present study was to describe a fast, simple diagnostic method for the differentiation of HES and secondary eosinophilic states. Beside the surface marker analysis of peripheral blood mononuclear cells (PBMC) we measured surface bound IgE molecules on lymphocytes and eosinophil cells, intracellular cytokines (IL-5, INFgamma) in CD4+ lymphocytes and eosinophil major basic protein (MBP) in eosinophils using flow cytometric detection method. The appearance of an IL-5 producing cell population with a decreased number of INFgamma positive lymphocytes was characteristic for the blood samples of HES patients. Predominance of Th2 cells with the appearance of a CD8+/CD3 /CD56+ cell population was restricted for the HES cases and could not be detected in secondary eosinophilic individuals. Our flow cytometric cytokine detection method (with parallel cell surface marker analysis) does not require cell separation or long term cell culture steps previously described for the detection of IL-5 producing cells. Therefore it seems to be a more appropriate approach for the differential diagnosis of primary and secondary eosinophilic states. PMID- 11882911 TI - Clot retention and spontaneous rupture with secondary pneumatosis of bladder wall following routine cystoscopy. AB - In this article we report an unusual case of spontaneous rupture of bladder wall following office-cystoscopy. It took place in a patient who suffered from low stage highgrade carcinoma of bladder with the different aggressive behavior. Finally, he underwent radical cystectomy, which showed micropapillary carcinoma and pneumatosis within the bladder wall. The cause of the latter finding is rather puzzling and has been never reported previously. PMID- 11882912 TI - GABA(B) receptor-mediated modulation of the firing pattern of ventral tegmental area dopamine neurons in vivo. AB - Previous work demonstrates the fundamental role of the firing pattern, specifically the burst firing mode of midbrain dopamine (DA) neurons in the regulation of DA release. Spontaneous burst firing has been shown to be dependent upon NMDA receptor activation of the DA cells. In addition to NMDA receptors, previous studies have reported that also GABA(B) receptors modulate the firing pattern of DA neurons in the substantia nigra. In the present electrophysiological study the role of GABA(B) receptors in the modulation of the firing pattern of DA neurons in the ventral tegmental area (VTA) in anaesthetised Sprague-Dawley rats was analysed. Systemic administration of the selective and potent GABA(B) receptor agonist baclofen dose-dependently reduced firing rate and burst firing in VTA DA neurons. An increase in the regularity of DA cell firing was also observed. All these effects were effectively antagonized by administration of the selective GABA(B) antagonist CGP 35348 (100 mg/kg or 200 mg/kg, i.v.). Administration of CGP 35348 (400 mg/kg, i.v.) per se was associated with a long-lasting increase in burst firing activity. The effects of systemic administration of baclofen, alone or in combination with CGP 35348, on the firing rate were largely mimicked by local microiontophoretic application of the drugs onto the DA neurons.Our findings indicate that central GABA(B) receptors may contribute to control of the burst firing mode of VTA DA neurons. Physiologically, activation of GABA(B) receptors may subserve a dampening function on VTA DA cell excitability which may counterbalance NMDA receptor mediated excitation. PMID- 11882913 TI - Essential role of ICAM-1/LFA-1 interaction in synergistic effect of IL-18 and IL 12 on IFN-gamma production in human PBMC. AB - IL-18 (0-100 ng/ml) specifically upregulated ICAM-1 expression on monocytes in human PBMC as demonstrated in our previous study. In the present study, we examined whether the synergistic upregulation of ICAM-1 occurred after the stimulation with the combination of IL-18 and IL-12 and whether the synergistic production of IFN-gamma was dependent on the interaction between ICAM-1 on monocytes and LFA-1 on NK/T cells. The effect of IL-12 on ICAM-1 expression on monocytes was marginal even at the highest concentration (100 ng/ml). However, in the presence of IL-12 (100 ng/ml), the expression of ICAM-1 induced by IL-18 was significantly enhanced as compared with that obtained by IL-18 alone. In addition to the expression of ICAM-1 on monocytes, IFN-gamma production was synergistically stimulated by IL-18 and IL-12. Anti-ICAM-1 and anti-LFA-1 Abs exhibited significant inhibitory effect on enhanced production of IFN-gamma by the combination of two cytokines, in particular, anti-ICAM-1 showing the complete inhibition. These results as a whole indicated that synergistic effect of IL-18 and IL-12 on IFN-gamma production in human PBMC is ascribed to the synergism of the effect of two cytokines on ICAM-1 expression on monocytes and that the subsequent ICAM-1/LFA-1 interaction plays an important role in the enhanced production of IFN-gamma. PMID- 11882914 TI - Relaxant effect of capsazepine in the isolated rat ileum. AB - We have evaluated the effect of the vanilloid receptor agonists resiniferatoxin (RTX), capsaicin and piperine and of the vanilloid receptor antagonist capsazepine on the resting tone in the isolated rat ileum. Capsazepine (10(-8)-3 x 10(-5) M) produced a concentration-related relaxation (8 +/-3%-49 +/-3%) of the rat ileum. By contrast RTX (up to 10(-8) M), capsaicin (up to 10(-6) M) and piperine (up to 10(-5) M) were without effect. Pre-treatment with capsaicin [either in vivo (50 mg/kg s.c.) or in vitro (10(-6) M)] did not modify the inhibitory effect of capsazepine. The L-type Ca2+ channel antagonist nifedipine (10(-6) M), but not the N-type Ca2+ channel antagonist omega-conotoxin GVIA (3 x 10(-8) M) nor the Na+ channel blocker tetrodotoxin (3 x 10(-7) M), counteracted the inhibitory effect of capsazepine. The NK1 receptor antagonist SR 140333 (10( 7) M), the NK2 receptor antagonist SR 48968 (10(-6) M), the NK3 receptor antagonist SR 142801 (10(-7) M), atropine (10(-6) M), hexamethonium (10(-4) M), phentolamine (10(-6) M) plus propranolol (10(-6) M), N(G)-nitro- L-arginine methyl ester ( L-NAME 3 x 10(-4) M), apamin (10(-7) M), methysergide (10(-6) M), the calcitonin gene-related peptide (CGRP) antagonist hCGRP 8-37 (1.5 x 10(-6) M), the VIP antagonist hGRF 1-29 (10(-5) M) did not modify the inhibitory effect of capsazepine. Capsazepine (2.5-40 mg/kg) also decreased upper gastrointestinal transit in vivo. It is concluded that the vanilloid antagonist capsazepine has a direct relaxing effect on rat intestinal smooth muscle which could involve L-type calcium channels. We found no evidence to suggest that capsazepine is antagonizing an endogenous vanilloid. PMID- 11882915 TI - Rat NKCC2/NKCC1 cotransporter selectivity for loop diuretic drugs. AB - It is generally assumed that bumetanide possesses some selectivity for the renal Na-K-Cl cotransporter NKCC2, although the results are scarce in the literature and comparisons were done with extra-renal NKCC1 at its basal, almost silent state. Here we investigated NKCC2/NKCC1 selectivity of loop diuretic drugs (bumetanide, piretanide and furosemide) as a function of the NKCC1 activated state (NKCC1 was activated by hypertonic media). NKCC2 activity was measured in isolated rat medullary thick ascending limb (mTAL) and NKCC1 in rat thymocytes and erythrocytes. When NKCC2 was compared with NKCC1at its activated state, all three diuretic drugs inhibited NKCC2 and NKCC1 with the same potency (bumetanide pIC50=6.48, 6.48 and 6.47; piretanide pIC50=5.97, 5.99 and 6.29; and furosemide pIC50=5.15, 5.04 and 5.21 for mTAL NKCC2, erythrocyte NKCC1 and thymocyte NKCC1, respectively). Basal NKCC1 exhibited a lower diuretic sensitivity, although with marked differences depending on the diuretic drug and the cell type in consideration and with the notable exception of furosemide in erythrocytes. Molecular modelling showed that bumetanide and piretanide possess four potentially active groups, of which three are shared with furosemide at similar intergroup distances. Of these three common groups, one should not bind to basal NKCC1 in thymocytes. The fourth (phenoxy) group (absent in furosemide) confers higher lipophilicity and should not bind to basal NKCC1 in erythrocytes. In conclusion, loop diuretics had no NKCC2/NKCC1 selectivity, when NKCC1 is measured at its activated state. Basal NKCC1 has a reduced diuretic sensitivity, of very different magnitude depending on the diuretic drug and cell type in consideration. PMID- 11882916 TI - Somatostatin (SRIF) modulates distinct signaling pathways in rat pituitary tumor cells; negative coupling of SRIF receptor subtypes 1 and 2 to arachidonic acid release. AB - The somatotropin release-inhibiting factor somatostatin-14 (SRIF) is known to activate distinct receptor subtypes (sst1-5). In rat pituitary tumor cells (GC cells), sst2 but not sst1 receptors mediate the SRIF-induced inhibition of intracellular concentration of Ca2+ ([Ca2+]i) and are negatively coupled to cAMP dependent pathways. In the present study, transduction mechanisms coupling distinct SRIF receptors to their specific functional role were investigated with the use of both SRIF agonists with well-known affinity at individual SRIF receptors and the sst2 receptor antagonist L-Tyr(8) isomer of Cyanamid 154806 (CYN-154806). Our results demonstrate that sst1 and sst2 receptors are coupled to distinct signaling pathways in GC cells. In particular, sst2 receptors are negatively coupled to the cAMP-dependent pathway and this pathway is partially responsible for the sst2 receptor-mediated inhibition of [Ca2+]i. In addition, sst1 and sst2 receptors are both coupled to a decrease of arachidonic acid (AA) release with an efficacy similar to that of SRIF, suggesting that SRIF reduces AA release through either a partial activation of both receptors or the activation of one at a time. This finding is important given the well-accepted role for phospholipase A2 (PLA2) as a positive signaling component in transduction pathways of SRIF receptors. sst1 and sst2 receptor negative coupling to PLA2/AA pathways does not seem to be implicated in the SRIF-induced inhibition of [Ca2+]i. The possible role for the SRIF-mediated inhibition of AA release in GC cell function remains to be elucidated. PMID- 11882917 TI - Supersensitivity of 5-HT1A autoreceptors and alpha2-adrenoceptors regulating monoamine synthesis in the brain of morphine-dependent rats. AB - The sensitivity of 5-HT1A serotonin receptors and alpha2-adrenoceptors (autoreceptors and heteroreceptors) modulating brain monoamine synthesis was investigated in rats during morphine treatment and after naloxone-precipitated withdrawal. The accumulation of 5-hydroxytryptophan (5-HTP) and 3,4 dihydroxyphenylalanine (DOPA) after decarboxylase inhibition was used as a measure of the rate of tryptophan and tyrosine hydroxylation in vivo. Acute morphine (3-100 mg/kg, 1 h) increased the synthesis of 5-HTP/5-HT in various brain regions (15%-35%) and that of DOPA/dopamine (DA) in striatum (28%-63%), but decreased the synthesis of DOPA/noradrenaline (NA) in hippocampus and cortex (20% 33%). Naloxone (2-60 mg/kg, 1 h) did not alter the synthesis of 5-HTP or DOPA in brain. Tolerance to the inhibitory effect of morphine on DOPA/NA synthesis and a sensitization to its stimulatory effects on DOPA/DA and 5-HTP/5-HT synthesis were observed after chronic morphine and/or in morphine-withdrawn rats. In morphine dependent rats (tolerant and withdrawn states) the inhibitory effects of the 5 HT1A agonists 8-OH-DPAT and buspirone (0.1 mg/kg, 1 h), and that of the alpha2 adrenoceptor agonist clonidine (0.1 mg/kg, 1 h), on the synthesis of 5-HTP/5-HT were potentiated (25%-50%). Moreover, the effect of 8-OH-DPAT was antagonized by WAY 100135, a selective 5-HT1A antagonist. In morphine-dependent rats (tolerant state), the inhibitory effects of clonidine on the synthesis of DOPA/NA (hippocampus, hypothalamus) and DOPA/DA (striatum) also were potentiated (35% 55%). In summary, we conclude that morphine addiction is associated with supersensitivity of 5-HT1A serotonin receptors and alpha2-adrenoceptors (autoreceptors and heteroreceptors) that modulate the synthesis of monoamines in brain. PMID- 11882918 TI - KCO912: a potent and selective opener of ATP-dependent potassium (K(ATP)) channels which suppresses airways hyperreactivity at doses devoid of cardiovascular effects. AB - ATP-sensitive potassium (K(ATP)) channel openers can obviate experimental airways hyperreactivity (AHR) and have shown therapeutic benefit in asthma. However, the clinical potential of such compounds has been compromised by cardiovascular side effects. We report here the pharmacological properties of (3 S,4 R)-3,4-dihydro-3 hydroxy-2,2-dimethyl-4-(2-oxo-1-piperidinyl)- N-phenyl-2 H-1-benzopyran-6 sulphonamide (KCO912), a K(ATP) channel opener which suppresses AHR at doses devoid of cardiovascular effects.Specific interaction of KCO912 with the native vascular channel and the sulphonylurea receptor subunit (SUR2B) of the vascular K(ATP) channel was shown in radioligand binding assays. In rat aortic strips, KCO912 inhibited specific binding of [3H]P1075 and [3H]glibenclamide with up to 100% efficacy and with p Ki values of 8.28 and 7.96, respectively. In HEK cells transfected with the recombinant vascular K(ATP) channel (Kir6.1 + SUR2B), the compound elicited a concentration-dependent outward current (pEC50 6.8) and in preloaded rat aortic rings it induced a concentration-dependent glibenclamide sensitive 86Rb+ efflux (pEC50 7.51). Following intratracheal (i.t.) administration of KCO912 to guinea pigs, AHR induced by immune complexes or ozone was rapidly (<5 min) reversed (ED50 values 1 microg/kg and 0.03 microg/kg, respectively). Changes in blood pressure were seen only at doses =100 microg/kg yielding 'therapeutic ratios' of 100 and 3333, respectively. In addition, KCO912 reversed AHR induced by lipopolysaccharide (LPS; ED50 0.5 microg/kg i.t.) and a dose of 1 microg/kg i.t. fully reversed AHR induced by subchronic treatment with salbutamol. At doses which suppressed AHR, KCO912 had no anti-bronchoconstrictor effects in normoreactive guinea pigs. In spontaneously hyperreactive rhesus monkeys, KCO912, given by inhalation, inhibited methacholine-induced bronchoconstriction (ED50 1.2 microg/kg) but had no significant effects on blood pressure or heart rate at all doses tested (therapeutic ratio >100). In rats given 3 mg/kg of KCO912 by inhalation, the ratio of the area under the concentration-time curve (AUC) for lung to the AUC in blood was 190 and the compound was rapidly cleared (initial t1/2 approximately 30 min). Thus, the wide therapeutic window following administration of KCO912 to the lung seems likely to reflect slow or incomplete passage of KCO912 from the lung into the systemic circulation coupled with rapid removal from the systemic circulation.Thus, when given locally to the airways in both guinea pigs and monkeys, KCO912 suppresses AHR at doses devoid of cardiovascular effects and has a significantly better therapeutic window than representative earlier generation K(ATP) channel openers defined in the same models. Given the pivotal role of AHR in the pathophysiology of asthma and the preclinical profile of KCO912, this compound was selected for clinical evaluation. PMID- 11882919 TI - G-protein betagamma-subunits contribute to the coupling specificity of the beta2 adrenergic receptor to G(s). AB - Receptors and heterotrimeric G-proteins interact with a high degree of specificity, the molecular basis of which is only partially understood. In the present study, we analyzed the influence of different G-protein betagamma subunits on the coupling of the beta2-adrenergic receptor to G(s). Sf9-cells were infected with baculoviruses coding for the beta2-adrenergic receptor, alpha(s,Short) or alpha(s,Long), and various beta- and gamma-subunits. The ability of different beta- and gamma-subunits to correctly dimerize was assessed by limited proteolysis of proteins expressed in Sf9-cells and additionally by analysis of beta/gamma-interaction in the yeast two-hybrid system. Agonist induced GTPgammaS-binding to alpha(s,Short)beta(1)gamma-trimers was significantly higher than to alpha(s,Short)beta2gamma-combinations, when gamma4, gamma5, or gamma7 were co-expressed. Because beta(5) did not support coupling of the beta(2) adrenergic receptor to G(s), the 87 C-terminal amino acids of Gbeta(5) assumed to encompass the beta-subunit interface with the receptor were substituted by the corresponding sequence of beta(1). Whereas this beta(5)/beta(1)-chimera did not promote GTPgammaS-binding to alpha(s), histamine H(1)-receptor-dependent GTPgammaS-binding to alpha(q) was supported by this chimeric beta-subunit and by wild-type beta(5). Our findings argue that the betagamma-subunit composition contributes directly to the specificity of beta(2)-adrenergic receptor-mediated G(s)-activation. PMID- 11882920 TI - Characterization of phospholipase C activity at h5-HT2C compared with h5-HT2B receptors: influence of novel ligands upon membrane-bound levels of [3H]phosphatidylinositols. AB - Employing a novel, rapid and sensitive method for evaluation of phospholipase C (PLC) activity, the present study characterized the actions of diverse agonists and antagonists at human (h)5-HT2C receptors expressed in Chinese Hamster Ovary (CHO) cells. In addition, affinities and efficacies at these sites were compared with those obtained at h5-HT2B receptors.5-HT elicited a robust and rapid reduction in levels of the pre-labelled, membrane-bound substrate of PLC, [3H]phosphatidylinositols ([3H]PI). The time-course of [3H]PI depletion paralleled that of [3H]inositol phosphate ([3H]IP) accumulation, as determined by conventional anion exchange chromatography. Inactivation of h5-HT2C receptors with the alkylating agent, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), revealed a large receptor reserve, with half-maximal PLC activation induced by a concentration of 5-HT occupying only 5% of sites. In analogy to 5-HT ( Emax=100%), DOI, MK212 and mCPP, as well as the novel ligands, Ro600332, Ro600175 and BW723C86, showed "full" efficacy at h5-HT2C sites. Their efficacies were similar at h5-HT2B sites, with the exception of mCPP and MK212, which acted as partial agonists. Further, lisuride and Ro600869 behaved as partial agonists and antagonists at h5-HT2C and h5-HT2B receptors, respectively. As concerns functional selectivity (potency for induction of [3H]PI depletion), only Ro600175 preferentially activated h5-HT2B sites. In contrast, Ro600332 preferentially activated h5-HT2C receptors. Amongst antagonists, RS102221 and SB242084 displayed a marked preference for h5-HT2C sites, whereas LY266097, S33526 and SB204741 behaved as selective antagonists at h5-HT2B receptors. At both h5-HT2C and h5 HT2B receptors, antagonist potency (p Kb) and binding affinity (p Ki) were highly correlated. In conclusion, this rapid and innovative method for determination of PLC activity permitted characterization of an extensive range of novel ligands at h5-HT2C receptors. Although several antagonists clearly differentiated h5-HT2C from h5-HT2B receptors under these conditions, highly selective agonists remain to be identified. PMID- 11882921 TI - Discharge behavior of motor units in knee extensors during the initial stage of constant-force isometric contraction at low force level. AB - To elucidate the strategy of the activity of motor units (MUs) to maintain a constant-force isometric contraction, I examined the behavior of MUs in knee extensor muscles [(vastus medialis (VM), vastus lateralis (VL) and rectus femoris (RF)] during a sustained contraction at 5% of maximal voluntary contraction for 5 min. In all cases, the spike interval exhibited an elongating trend, and two discharge patterns were observed, continuous discharge and decruitment. In continuous-discharge MUs, the trend slope was steep immediately after the onset of constant force (steep phase), and then became gentle (gentle phase). Decruitments were observed frequently during each phase, and additional MU recruitment was observed throughout the contraction. The mean value of recruitment threshold force did not differ among the extensors. The mean spike interval at the onset of constant-force isometric contractions was shorter in RF than in VL. However, there were no differences in the duration and extent of the elongating trend, decruitment time and recruitment time among the extensors. The electromyogram of the antagonist biceps femoris muscle revealed no compensatory change for extensor activity. These results indicated that at a low force level, the strategy employed by the central nervous system to maintain constant force appears to involve cooperation among elongating trends in the spike interval, decruitment following elongation, and additional MU recruitment in synergistic muscles. PMID- 11882922 TI - Seasonal variation of alterations in exercise-induced body composition in obese Japanese women. AB - Although many investigations have been conducted to determine the effects of exercise and/or diet prescription, seasonal variations in weight loss have not been thoroughly investigated in Japan. The present investigation was undertaken to determine seasonal variations in body composition and anthropometric characteristics during a weight-loss program. One hundred and twenty-seven women [mean (SD) age 44.3 (11.7) years] participated in this study. The subjects were categorized into four groups based on the season in which the weight-loss program was begun: subjects who joined our exercise program in winter (group WE), subjects who joined in summer (group SE), and two control groups, one for winter and one for summer (group WC and group SC, respectively). The subjects of groups WE and SE completed a 3-month exercise and food-restriction program. Analyses of covariance revealed that the subjects of group WE seemed to have attained more desirable changes. This group showed a decrease in body mass (-1.9 (1.5) kg) and percent body fat (-4.6 (4.8)%), while an increase in fat-free mass (1.6 (3.2) kg) during the experimental period. Our data suggest that the better season for desirable weight loss in Japan is winter, compared to summer. These alterations may be attributable, at least in part, to the fact that Japanese people are generally acclimatized to a cold climate in this season. The mechanisms involved in our speculation have not been well validated, but it seems clear that a significant difference in dietary intake might have affected the results of this study. PMID- 11882923 TI - Time-of-day effect on nonthermal control of sweating response to maintained static exercise in humans. AB - To investigate the influence of nonthermal factors in the time-of-day effect on the sweating response to maintained static exercise, eight healthy male subjects performed handgrip exercise at 20%, 35% and 50% maximal voluntary contraction (MVC) for 60 s at 0600 hours (morning) and at 1800 hours (evening). Oesophageal temperature ( T(oes)) before the experiment showed a diurnal rhythm [mean (SEM)] [36.3 (0.1) (morning) compared to 36.8 (0.1) degrees C (evening), P<0.01]. Experiments were conducted with subjects in a state of mild hyperthermia during which the mean skin temperature ( T(sk)) was kept constant at 35.5-36.5 degrees C using a water-perfused suit to activate sudomotor responses. The T(oes) and mean T(sk)remained stable during the pre-exercise, handgrip exercise and recovery periods. The response in sweating rate (DeltaSR) on the chest and forearm to handgrip exercise increased significantly with increasing exercise intensity in both the morning and evening tests ( P<0.05). The DeltaSR on the palm did not change significantly with increasing exercise intensity in the morning test ( P>0.1). During handgrip exercise at 50% MVC only, DeltaSR on the chest, forearm and palm in the evening was significantly higher than in the morning ( P<0.05). On the other hand, mean arterial blood pressure and the rating of perceived exertion during 50% MVC handgrip exercise were not significantly different between the morning and evening ( P>0.1). These results indicate the presence of a time-of-day effect on nonthermal control of the sweating response to isometric handgrip exercise, and that this effect is dependent on exercise intensity. PMID- 11882924 TI - Skin blood flow in the human hand in relation to applied pressure. AB - Our aim was to study the relationship between external pressure and skin blood flow (BF) in the area of the palm (P) of the human hand and the relationship between BF and pressure pain thresholds. A special probe was designed for measuring simultaneously the skin BF using a laser Doppler flowmeter and the pressure using an algometer. The normal BF in the distal phalanx of the index finger and in the middle of the thenar musculature was higher than in the middle phalanx of the middle finger and the middle of P. A pressure of 15-33 kPa was required to reduce BF by 50% and 30-52 kPa to reduce by 85% except in P where the pressure had to be much higher; for 50% of the subjects the reduction in BF did not reach 85% even at 100 kPa. After the removal of the pressure the BF was 170% 230% of the normal readings. The BF did not correlate with the pressure pain thresholds. These results indicate that continuously applied pressure in the hand should not be above 50 kPa to avoid tissue damage due to the cessation of BF in the skin. PMID- 11882925 TI - Hyperbaric oxygen and neutrophil accumulation/tissue damage during permanent focal cerebral ischaemia in rats. AB - The use of hyperbaric oxygenation (HBO) for the treatment of severe brain ischaemia remains controversial. The HBO may interfere with destructive neutrophil (PMN) infiltration following ischaemia/reperfusion. The effects of HBO on PMN accumulation and the area of ischaemic tissue damage were investigated in rats having permanent focal ischaemia (4 h). The right middle cerebral arteries of a group of Wistar rats were permanently occluded. The rats were then randomly divided into those ( n=7) to be treated with HBO at 2 atm for 230 min and those ( n=8) to breathe air at atmospheric pressure for an equivalent period. The HBO had no effect on permanent ischaemia, as there was no significant difference in the area of ischaemic tissue damage between HBO-treated [mean (SD)] [331 (88) mm(3)] and non-treated animals [322 (111) mm(3)]. Moreover, the increase in myeloperoxidase [5.4 (4.1) compared to 2.4 (1.2) pg x g(-1) wet weight of brain] was not significantly different. The results indicate that HBO did not reduce tissue damage during 4 h of permanent focal ischaemia. PMID- 11882926 TI - Circulating transforming growth factor beta1 (TGFbeta1) is elevated by extensive exercise. AB - Transforming growth factor beta1 (TGFbeta1) is a multifunctional growth factor involved in immune function, atherosclerosis, fibrotic disease, diabetic complications and bone turnover. It is synthesized in large quantities by bone cells in response to hormones and mechanical stimuli. Plasma contains inactive "latent" TGFbeta1, which consists of the precursor molecule and a TGFbeta1 binding protein. Platelets store latent TGFbeta1 in their alpha-granules, and serum therefore contains large amounts of latent TGFbeta1. We developed a technique for determining latent plasma TGFbeta1 and investigated whether circulating TGFbeta1 is affected by the stimulation of bone formation in response to strength training. Ten healthy students with low training activity participated in a heavy exercise programme over 4 weeks. Blood was drawn into citrate-filled syringes containing prostaglandin E(1) (PGE(1)) and immediately centrifuged at 4 degrees C. TGFbeta1 was determined with a sandwich ELISA standardized with National Institute for Biological Standards and Controls (NIBSC) materials. Six of the ten students completed the training. Highly reproducible values (500-600 pg/ml) of latent TGFbeta1 in plasma were determined. Baseline levels of TGFbeta1 were 525 (50) pg/ml [mean (SE)], which is in the range observed for young adults. TGFbeta1 concentrations rose significantly to 710 (65) pg/ml after 2 weeks of training and thereafter slowly declined to 650 (62) pg/ml after 2 weeks and 440 (33) pg/ml after 4 weeks, respectively. No active TGFbeta1 was detectable in citrate PGE1 plasma samples. Serum levels were between 6000 and 10,000 pg/ml and contained 200-400 pg/ml active TGFbeta1. In contrast to previous reports, plasma did not contain measurable amounts of circulating active TGFbeta1. We demonstrate that heavy exercise transiently elevates latent TGFbeta1 concentrations in plasma. TGFbeta1 is produced by osteoblasts in considerable amounts; therefore, we assume that the observed changes are partly due to enhanced TGFbeta1 production or release in bone, since the quantities of TGFbeta1 produced by other cells are comparably small. PMID- 11882927 TI - Effect of an acute period of resistance exercise on excess post-exercise oxygen consumption: implications for body mass management. AB - Studies have shown metabolism to remain elevated for hours following resistance exercise, but none have gone beyond 16 h, nor have they followed a whole body, high intensity exercise protocol. To examine the duration of excess post-exercise oxygen consumption (EPOC) following a period of heavy resistance exercise, seven healthy men [mean (SD) age 22 (3) years, height 177 (8) cm, mass 83 (10) kg, percentage body fat 10.4 (4.2)%] engaged in a 31 min period of resistance exercise, consisting of four circuits of bench press, power cleans, and squats. Each set was performed using the subject's own predetermined ten-repetition maximum and continued until failure. Oxygen consumption ( ) measurements were obtained at consistent times (34 h pre-, 29 h pre-, 24 h pre-, 10 h pre-, 5 h pre , immediately post-, 14 h post-, 19 h post-, 24 h post-, 38 h post-, 43 h post-, and 48 h post-exercise). Post-exercise measurements were compared to the baseline measurements made at the same time of day. The was significantly elevated ( P<0.05) above baseline values at immediately post, 14, 19, and 38 h post exercise. Mean daily values for both post-exercise days were also significantly elevated above the mean value for the baseline day. These results suggest that EPOC duration following resistance exercise extends well beyond the previously reported duration of 16 h. The duration and magnitude of the EPOC observed in this study indicates the importance of future research to examine a possible role for high intensity resistance training in a weight management program for various populations. PMID- 11882928 TI - Individual factors and physical work load in relation to sensory thresholds in a middle-aged general population sample. AB - The aim of this study was to explore individual and occupational factors possibly related to sensory thresholds in 484 middle-aged men and women from the general population. Multivariate analyses were performed using a linear model including eight covariates (age, body height, skin temperature, smoking habits, musculoskeletal symptoms during the last week before examination, isometric muscle strength, aerobic capacity, and average physical work load during the last 15 years). Men and women were considered separately in all analyses. Major findings were as follows: pressure pain thresholds increased with muscle strength; cold perception thresholds on the foot improved with increasing skin temperature; vibration and warm perception thresholds on the foot increased with age and body height; pressure pain threshold on the leg was decreased and vibration threshold on the hand was increased in women with musculoskeletal symptoms; pressure pain thresholds were slightly increased in men reporting long lasting high physical work load. Significant sex differences were found for the majority of sensory thresholds, that is higher thresholds in men. However, body height was found to be more important than gender for differences in vibration and warm perception thresholds on the foot. PMID- 11882929 TI - Acute effects of stretching on the neuromechanical properties of the triceps surae muscle complex. AB - Previous research has shown that an acute bout of passive muscle stretching can diminish performance in certain movements where success is a function of maximal force and/or power output. Two possible mechanisms that might account for such findings are a change in active musculotendinous stiffness and a depression of muscle activation. To investigate the likelihood of these two mechanisms contributing to a post-stretch reduction in performance, we examined the acute effects of stretching on the active stiffness and muscle activation of the triceps surae muscle group during maximal single-joint jumps with movement restricted to the ankle joint. Ten males performed both static (SJ) and countermovement (CMJ) jumps before and after passively stretching the triceps surae. Electrical activity of the triceps surae during each jump was determined by integrating electromyographic recordings (IEMG) over the course of the movement. Triceps surae musculotendinous stiffness was calculated before and after stretching using a technique developed by Cavagna (1970). Following stretching, a significant decrease [mean (SD) 7.4 (1.9)%; P<0.05] in jump height for the CMJ occurred, but for the SJ, no significant ( P>0.05) change in jump height was found. A small but significant decrease [2.8 (1.24)%; P<0.05] in stiffness was noted, but the magnitude of this change was probably not sufficient for it to have been a major factor underlying the decline in CMJ performance. Paradoxically, after stretching, the SJ exhibited a significant ( P<0.05) decrease in IEMG, but the IEMG for the CMJ remained unchanged ( P>0.05). It appears that an acute bout of stretching can impact negatively upon the performance of a single-joint CMJ, but it is unlikely that the mechanism responsible is a depression of muscle activation or a change in musculotendinous stiffness. PMID- 11882930 TI - Validity and reliability of three commercially available breath-by-breath respiratory systems. AB - Information concerning the validity and reliability of commercial on-line gas analysis systems is limited. The aim of this study was to provide a comparison of the validity and reliability of three on-line systems (Oxycon Alpha, Oxycon Pro and Pulmolab EX670) with that of Douglas bags. Two separate studies were conducted. In study 1, the three gas analysis systems were compared with Douglas bags using a metabolic simulator over four increases in ventilation. In study 2, ten subjects were split into equal groups exercising at 100 W or 150 W for 85 min on three separate occasions. Each system was used twice per visit. Study 1 demonstrated that the Oxycon Alpha and Douglas bags produced similar respiratory values over all levels of ventilation. The Oxycon Pro tended to slightly overestimate mean expiratory flow (V(E) ), oxygen uptake ( VO2), carbon dioxide production (VCO2) and respiratory exchange ratio (RER) at the higher ventilations. The Pulmolab produced large overestimations at all ventilations for and RER (up to 26.3% away from expectations), whilst values for, and were slightly underestimated at higher ventilations (up to 7.5% from expectations). The results of study 2 support the findings of study 1, with the Oxycon Pro and Oxycon Alpha producing similar results compared to Douglas bags for, and RER. The coefficients of variation for and measured using Douglas bags, Oxycon Pro and Oxycon Alpha were 3.3-5.1%, 4.7-7.0% and 4.5-6.3%, respectively, whilst that for the Pulmolab was highly variable (26.8-45.8%). The exercise study showed the Oxycon Pro and Oxycon Alpha to be both valid and reliable on-line systems for the measurement of parameters of respiration, at least at workloads up to 150 W. PMID- 11882931 TI - Effects of prolonged low doses of recombinant human erythropoietin during submaximal and maximal exercise. AB - The aim of this study was to characterise the effect of prolonged low doses of recombinant erythropoietin (r-HuEPO) on the responses to submaximal and maximal exercise. Volunteer recreational athletes ( n=21) were divided into three groups: r-HuEPO+intravenous iron (EPO+IV, n=7), r-HuEPO+oral iron (EPO+OR, n=9) and placebo ( n=5). During the 12 week study, r-HuEPO or saline injections were given three times a week for the first 8 weeks and for the final 4 weeks the subjects were monitored but no injections were administered. The r-HuEPO doses were 50 IU x kg(-1) body mass for 3 weeks and 20 IU x kg(-1) body mass for the next 5 weeks. An exercise test comprising three submaximal intensities and then increments to elicit maximal aerobic power (VO2max ) was conducted during weeks 0, 4, 8 and 12. During week 0, the mean intensity of the submaximal stages was 60%, 72% and 81%. Blood taken at rest was analysed twice a week for haematocrit (Hct). The relative increases in at weeks 4, 8 and 12 were 7.7%, 9.7% and 4.5%, respectively, for the EPO+IV group; 6.0%, 4.7% and 3.1% for the EPO+OR group; and -0.5%, -0.1% and 1.0% for the placebo group, where the improvements at week 12 for the EPO+IV and EPO+OR groups remained significantly above week 0 values. The Hct was significantly elevated by 0.06 and 0.07 units at week 3 in the EPO+IV and EPO+OR groups, respectively, and was stable during the 5 weeks of low-dose r-HuEPO. After 8 weeks of r-HuEPO use, plasma lactate concentration tended to be lower at exercise intensities ranging from 60% to 100%. This study confirmed the ability of low doses of r-HuEPO to maintain Hct and at elevated levels. PMID- 11882932 TI - The physiological demands of sail pumping in Olympic level windsurfers. AB - This study investigated the physiological effects of sail pumping (PB)--a manoeuvre often adopted to provide additional propulsion to the board--in Olympic Class Windsurfing, following relaxation of the "no-pumping" rules by the International Federation. Fifteen Olympic-level windsurfers (10 men) from nine different countries volunteered for the study, which was performed during two international Olympic regattas. The measurements were carried out during actual sailing when both PB and not-pumping (NPB) using a portable metabolimeter. Windsurfing, when PB, elicited a dramatic increase in cardiorespiratory responses compared to NPB. Mean (SD) values for oxygen uptake and heart rate during NPB for the men and women were: 19.2 (4.4) and 15.7 (3.3) ml x kg(-1) x min(-1), and 110 (10) and 122 (12) beats x min(-1), respectively, whereas the values in PB were: 48.4 (5.7) and 40.2 (4.2) ml x kg(-1) x min(-1), and 165 (12) and 172 (13) beats x min(-1), respectively. All the PB parameters, with the exception of heart rate (HR), were significantly higher in the men than in the women but no differences were observed between the sexes in NPB with the exception of HR, which was higher in the women. Our results suggest sail pumping is as physically demanding as most aerobic sporting activities. In the context of the need to deal with a highly demanding athletic branch of sailing as part of an Olympic regatta, recommendations are made on how best to make physical and dietary preparations. PMID- 11882933 TI - Aspartic proteinases are expressed in pitchers of the carnivorous plant Nepenthes alata Blanco. AB - Carnivorous plants acquire significant amounts of nitrogen from insects. The tropical carnivorous plant Nepenthes accumulates acidic fluid containing aspartic proteinase (AP) in its trapping organs (pitchers), suggesting that the plant utilizes insect protein as a nitrogen source. Aspartic proteinases have been purified and characterized from sterile pitcher fluid of several species of Nepenthes; however, there is, as of yet, no information about sequence and expression of Nepenthes AP genes. To identify the pitcher AP, we cloned plant AP homologs from N. alata and examined their expressions. Five AP homologs ( NaAP1 NaAP5) were obtained by reverse transcription-polymerase chain reaction with degenerate primers designed for the conserved sequences of plant APs. Alignment of deduced amino acid sequences with other plant APs demonstrated that NaAP1 NaAP4 contained a plant-specific insert (PSI), a unique sequence of plant AP. However, NaAP5 did not possess the insert, and had a shorter sequence (by >100 amino acids) than the other APs. Northern analysis using a part of the coding region of NaAP1 as a probe showed that bands of approx. 1.8 kb corresponding to the sizes of NaAP1-NaAP4 mRNA were present in roots, stems, leaves, tendrils, and lower part of the pitchers, but a band of approx. 1.3 kb corresponding to the size of NaAP5 mRNA was not observed in any organs. In pitchers, highest expressions of NaAP1-NaAP4 were seen in the lower part of open pitchers containing natural prey, suggesting that the expressions of NaAP1-NaAP4 are coupled with prey capture. Transcripts of NaAP2 and NaAP4 were detected in the digestive glands, where AP secretion may occur. This result suggests that NaAP2 and NaAP4 are the possible APs secreted into the pitcher of N. alata. PMID- 11882934 TI - Random antisense cDNA mutagenesis as an efficient functional genomic approach in higher plants. AB - Most cellular processes in an organism depend on functions of expressed sequences. Thus, efficient large-scale functional assignment of expressed sequences is crucial for understanding cellular processes. Towards this goal in plants, we designed a "random antisense cDNA mutagenesis (RAM)" approach. In a pilot experiment, 1,000 transgenic plants of Arabidopsis thaliana (L.) Heynh. (ecotype Wassilevskija) expressing random antisense cDNA(s) were generated from Agrobacterium cultures harboring an Arabidopsis antisense cDNA library. We identified 104 mutant lines from the transgenic pool by visual screening. Genetic analysis suggested that 37% of the mutations were likely due to antisense effects. When the cDNA inserts were isolated from 11 mutant lines by polymerase chain reaction and reintroduced into plants to express the antisense transcripts, the original mutant phenotypes were reproduced in 7 cDNA clones. One of the cDNA clones did not generate a database match to any sequence with known functions, but did have a dramatic effect on the architecture of the inflorescence in the antisense transgenic plants. Through the RAM approach, it should be possible to assign a large number of expressed sequences to known in vivo functions in plants. PMID- 11882935 TI - The 7B-1 mutant in tomato shows blue-light-specific resistance to osmotic stress and abscisic acid. AB - Germination of wild-type (WT) tomato ( Lycopersicon esculentum Mill.) seed is inhibited by mannitol (100-140 mM) in light, but not in darkness, suggesting that light amplifies the responsiveness of the seed to osmotic stress (M. Fellner, V.K. Sawhney (2001) Theor Appl Genet 102:215-221). Here we report that white light (W) and especially blue light (B) strongly enhance the mannitol-induced inhibition of seed germination, and that the effect of red light (R) is weak or nil. The inhibitory effect of mannitol could be completely overcome by fluridone, an inhibitor of abscisic acid (ABA) biosynthesis, indicating that mannitol inhibits seed germination via ABA accumulation in seeds. The inhibition of WT seed germination by exogenous ABA was also amplified by W or B, but not by R. In a recessive, ABA-overproducing, 7B-1 mutant of tomato, seed germination and hypocotyl growth were resistant to inhibition by mannitol or exogenous ABA, both in W or B. Experiments with fluridone suggested that inhibition of hypocotyl growth by W or B is also partially via ABA accumulation. De-etiolation in the mutant was especially less in B compared to the WT, and there was no difference in hypocotyl growth between the two genotypes in R. Our data suggest that B amplifies the responsiveness of tomato seeds and hypocotyls to mannitol and ABA, and that W- or B-specific resistance of the 7B-1 mutant to osmotic stress or ABA is a consequence of a defect in B perception or signal transduction. PMID- 11882936 TI - Actomyosin promotes cell plate alignment and late lateral expansion in Tradescantia stamen hair cells. AB - Cytokinesis in higher-plant cells involves the formation of a cell plate in the interzone between the separating chromatids. The process is directed by the phragmoplast, an array of microtubules, actin filaments, and membranous elements. To determine if the role of actin in cytokinesis is dependent on myosin, we treated stamen hair cells of Tradescantia virginiana L. with 2,3-butanedione monoxime (BDM), an inhibitor of myosin ATPase and ML-7, a specific inhibitor of myosin light-chain kinase. Treatment with BDM resulted in a tilted cytokinetic apparatus during early initiation and a wavy cell plate with curved phragmoplasts during late lateral expansion. Treatment with ML-7 also resulted in inefficient late lateral expansion of the cell plate, with effects ranging from slower expansion to complete inhibition. Taken together, these results implicate myosin in the control of cell plate expansion and alignment. PMID- 11882937 TI - ASYMMETRIC LEAVES1, an Arabidopsis gene that is involved in the control of cell differentiation in leaves. AB - During leaf development, the formation of dorsal-ventral and proximal-distal axes is central to leaf morphogenesis. To investigate the genetic basis of dorsoventrality and proximodistality in the leaf, we screened for mutants of Arabidopsis thaliana (L.) Heynh. with defects in leaf morphogenesis. We describe here the phenotypic analysis of three mutant alleles that we have isolated. These mutants show varying degrees of abnormality including dwarfism, broad leaf lamina, and aberrant floral organs and fruits. Genetic analysis revealed that these mutations are alleles of the previously isolated mutant asymmetric leaves1 ( as1). In addition to the leaf phenotypes described previously, these alleles display other phenotypes that were not observed. These include: (i) some rosette leaves with petiole growth underneath the leaf lamina; (ii) leaf vein branching in the petiole; and (iii) a leaf lamina with an epidermis similar to that on the petiole. The mutant phenotypes suggest that the ASYMMETRIC LEAVES1 ( AS1) gene is involved in the control of cell differentiation in leaves. As the first step in determining a molecular function for AS1, we have identified the AS1 gene using map-based cloning. The AS1 gene encodes a MYB-domain protein that is homologous to the Antirrhinum PHANTASTICA ( PHAN) and maize ROUGH SHEATH2 ( RS2) genes. AS1 is expressed nearly ubiquitously, consistent with the pleiotropic mutant phenotypes. High levels of AS1 expression were found in tissues with highly proliferative cells, which further suggests a role in cell division and early cell differentiation. PMID- 11882938 TI - Spatial distribution of the 26S proteasome in meristematic tissues and primordia of rice (Oryza sativa L.). AB - The 26S proteasome is known to play central roles in the growth of many eukaryotes. However, little is known regarding its distribution in higher plants. Here, we report the spatial distribution pattern of Rpn3 (a regulatory PA700 subunit) and C2 (a subunit of the 20S proteasome) in rice ( Oryza sativa L.) seedlings as determined by in situ hybridization. The transcripts were abundantly co-expressed in the apical and marginal meristems of shoots and roots. Interestingly, these transcripts also accumulated in the leaf and ligule primordia of the shoot apex. Our results suggest that the 26S proteasome is spatially distributed among various tissues and may be involved not only in cell division but also in organ formation in higher plants. PMID- 11882939 TI - Altered lignin structure and resistance to pathogens in spi 2-expressing tobacco plants. AB - The physiological role of the Norway spruce [ Picea abies (L.) Karst.] spi 2 gene, encoding a defense-related cationic peroxidase was examined in transgenic tobacco (Nicotiana tabacum L.). Expression of spi 2, under control of the 35S promoter, in tobacco plants resulted in higher total peroxidase activities. The phenotype of the spi 2-transformed lines was normal. The spi 2-transformed lines displayed lignin levels similar to levels in the control line, but with some alteration in lignin histochemistry and structure. These changes were associated with reduced flexibility of the tobacco stems. The defense against pathogenic microorganisms was altered in the transgenic tobacco plants compared with control plants. High peroxidase activities increased the susceptibility to the pathogenic oomycete Phytophthora parasitica var. nicotianae, but increased the ability of the tobacco plants to suppress growth of the pathogenic bacterium Erwinia carotovora. PMID- 11882940 TI - A fruit-specific phospho enolpyruvate carboxylase is related to rapid growth of tomato fruit. AB - Malic and citric acids accumulate in cherry tomato (Lycopersicon esculentum Mill.) fruit during the period of rapid growth, from the end of cell division to the onset of ripening. The involvement of phospho enolpyruvate carboxylase (PEPCase, EC 4.1.1.31) in organic acid accumulation and tomato fruit development was investigated. Two PEPCases, named LYCes;Ppc1 and LYCes;Ppc2 and mapped to chromosomes 12 and 7, respectively, were shown to be differentially expressed during tomato fruit development. LYCes;Ppc1 mRNA was present in all fruit tissues and in all other plant organs examined. In contrast, LYCes;Ppc2 was strongly and specifically expressed in fruit from the end of cell division to ripening. No LYCes;Ppc2 expression was detected by northern blot in other plant tissues. In fruit, the increase in LYCes;Ppc2 mRNA was closely followed by an increase in fruit PEPCase protein and activity, and was coincident with the increased accumulation of malate and citrate during the initial period of rapid growth rate, from 8 to 20 days post anthesis. Localization of LYCes;Ppc2 mRNA in young tomato fruit by in situ hybridization revealed that LYCes;Ppc2 is preferentially expressed in large cells of the pericarp and in enlarging cells of the gel surrounding the seeds. Examination of the kinetic and regulatory properties of the PEPCases of growing and ripening fruit further showed that PEPCase in growing fruit is less sensitive to low pH and malate inhibition, indicating a high phosphorylation state and/or the presence of a PEPCase isoform with these characteristics. Taken together, these results indicate that in developing tomato fruit PEPCase is probably important in permitting the synthesis of organic acids to provide the turgor pressure necessary for cell expansion. PMID- 11882941 TI - Benzoic acid biosynthesis in cell cultures of Hypericum androsaemum. AB - Biosynthesis of benzoic acid from cinnamic acid has been studied in cell cultures of Hypericum androsaemum L. The mechanism underlying side-chain shortening is CoA dependent and non-beta-oxidative. The enzymes involved are cinnamate:CoA ligase, cinnamoyl-CoA hydratase/lyase and benzaldehyde dehydrogenase. Cinnamate:CoA ligase was separated from benzoate:CoA ligase and 4-coumarate:CoA ligase, which belong to xanthone biosynthesis and general phenylpropanoid metabolism, respectively. Cinnamoyl-CoA hydratase/lyase catalyzes hydration and cleavage of cinnamoyl-CoA to benzaldehyde and acetyl-CoA. Benzaldehyde dehydrogenase finally supplies benzoic acid. In cell cultures of H. androsaemum, benzoic acid is a precursor of xanthones, which accumulate during cell culture growth and after methyl jasmonate treatment. Both the constitutive and the induced accumulations of xanthones were preceded by increases in the activities of all benzoic acid biosynthetic enzymes. Similar changes in activity were observed for phenylalanine ammonia-lyase and the xanthone biosynthetic enzymes benzoate:CoA ligase and benzophenone synthase. PMID- 11882942 TI - Detection and localization of pectin methylesterase isoforms in pollen tubes of Nicotiana tabacum L. AB - Pectin methylesterases (PMEs) were detected in tobacco ( Nicotiana tabacum) pollen tubes grown in vitro. Seven PME isoforms exhibiting a wide isoelectric point (pI) range (5.3-9.1) were found in crude extracts of pollen tubes. These isoforms were mainly retrieved in supernatants after low- and high-speed separation of the crude extract. Two isoforms, with pIs 5.5 and 7.3 and molecular weight about 158 kDa, were detected by immunoblotting with anti-flax PME antiserum. Localization of pectins and PME isoforms in pollen tubes was investigated by immunogold labelling with JIM5 monoclonal antibodies and anti flax PME antiserum, respectively. In germinated pollen grains, two PME isoforms were mainly detected in the exine, Golgi apparatus and secretory vesicles. In pollen tubes the same two PME isoforms were distributed along the outer face of the plasma membrane in the vicinity of the inner layer of the cell wall, in the Golgi and around secretory vesicles. In pollen grains, PME isoforms were, in some cases, mixed with acidic pectins in proximity to the outer surface of the plasma membrane. In pollen tubes the presence of PMEs inside secretory vesicles carrying esterified pectins supports the hypothesis that, during pollen tube growth, PMEs could be transferred by secretory vesicles in a precursor form and be activated at the tip where exocytosis takes place. PMID- 11882943 TI - The effect of exogenous sugars on the control of flux by adenosine 5' diphosphoglucose pyrophosphorylase in potato tuber discs. AB - The aim of this work was to investigate the effect of exogenous sugars on the extent to which starch synthesis in potato ( Solanum tuberosum L.) is controlled by adenosine 5'-diphosphoglucose pyrophosphorylase (EC 2.7.7.27; AGPase). Tuber discs were incubated in the presence of a range of concentrations of glucose and sucrose, and metabolic fluxes measured following the supply of [U-14C]glucose and measurement of the specific radioactivity of the hexose phosphate pool. In the presence of glucose there was a marked increase in the flux through glucose phosphorylating hexokinase, and at high concentrations of external glucose this led to a stimulation of the rate of starch and sucrose synthesis relative to those measured in the presence of sucrose. In the presence of glucose the ratio of the rate of starch synthesis to the rate of glycolysis was higher than in the presence of sucrose. Similar effects of glucose were observed at two stages of tuber development. We conclude that the presence of glucose perturbs the carbohydrate metabolism of tuber discs so that starch synthesis is favoured. In order to determine the extent to which AGPase controls flux, we measured fluxes in wild-type plants and transgenic plants with reduced AGPase activity as a result of the expression of a cDNA encoding the B subunit in the antisense orientation. In the presence of sucrose a reduction in AGPase activity had a greater impact on the rate of starch synthesis than in the presence of glucose. The flux control coefficient of AGPase over starch synthesis was higher in the presence of sucrose (0.7-0.9) than in the presence of glucose (0.4-0.6). Conversely, the impact of reduced AGPase activity on the rate of sucrose synthesis was lower in the presence of sucrose than glucose. In the presence of 200 mM sucrose the flux control coefficient of AGPase over the rate of sucrose synthesis was not significantly different from zero. This demonstrates that the nature of the sugar supplied to potato tuber discs can have a major influence on the distribution of control within metabolism. These data were also used to investigate the relationship between demand for ATP and the rate of hexose phosphate entry into glycolysis. A very strong correlation between ATP demand and glycolytic flux was demonstrated. PMID- 11882944 TI - Changes in oxidative processes and components of the antioxidant system during tomato fruit ripening. AB - Analysis of the oxidative processes taking place during fruit ripening in a salad tomato variety (Lycopersicon esculentum Mill. cv. Ailsa Craig) revealed changes in oxidative and antioxidative parameters. Hydrogen peroxide content, lipid peroxidation and protein oxidation were measured as indices of oxidative processes and all were found to increase at the breaker stage. The levels of the aqueous-phase antioxidants, glutathione and ascorbate, increased during the ripening process and these increases were associated with significant changes in their redox status, becoming more reduced as ripening progressed. Changes in the activities of superoxide dismutase, catalase and the enzymes involved in the ascorbate-glutathione cycle during ripening indicated that the antioxidative system plays a fundamental role in the ripening of tomato fruits. PMID- 11882945 TI - Potassium drives daily reversible thallus enlargement in the marine red alga Porphyra leucosticta (Rhodophyta). AB - Growth rate in terms of area expansion per 30 min was measured in the marine red algae Porphyra leucosticta under light/dark cycles of 6:6 h. Thalli grown in artificial seawater (ASW), under controlled ionic concentrations, showed a rapid thallus expansion just after light-on (morning-peak). Dark phase began with a significant thallus contraction (dark-peak) but no growth was observed until the next light phase. The removal of K+ from the medium inhibited the reversible growth peak that this species shows after light-on. On the other hand, the removal of Na+ did not have an apparent effect on growth pattern. Addition of Rb+ to K+-free ASW restored the morning-peak to 60% of its value in ASW, but addition of Li+ failed to restore the morning-peak. Intracellular ion analyses revealed that after light-on, the internal K+ content of the cells of this species increased 4-fold in 90 min, reaching an intracellular concentration of up to 300 micromol K+ per gram fresh weight, and that this value remained fairly constant over the light phase. Addition of 100 mM of tetraethylammonium, a specific K+ channel blocker, inhibited the morning-peak by 40%. The Na+ and Cl- contents in the cells increased rapidly in the first 45 min of the light phase, but then the internal concentrations of both ions decreased to their minimum values in the light phase. The carbonic anhydrase inhibitors acetazolamide and 6-ethoxyzolamide did not affect thallus expansion during the light phase. In contrast, thallus expansion in the light phase was completely inhibited by diethylstilbestrol, an inhibitor of the putative primary pump, and by 3-(3',4'-dichlorophenyl)-1,1 dimethylurea (DCMU), an inhibitor of photosynthesis, but not by sodium vanadate, a specific inhibitor of the plasma-membrane H+-ATPase. We suggest that the periodic oscillation of the short-term growth rate of P. leucosticta occurs in response to potassium fluxes, which control the osmotic pressure and eventually the relative cell volume. The possible effects of the loosening of the cell wall and the internal K+ concentration on the growth of this species are also discussed. PMID- 11882946 TI - Characterisation of the oxygen fluxes in the division, elongation and mature zones of Vitis roots: influence of oxygen availability. AB - Oxygen fluxes into and from root cells of Vitis rupestris (flooding sensitive), V. riparia (flooding tolerant) and V. vinifera (medium tolerance to flooding) were measured under different levels of O2 availability using a recently developed polarographic O2-selective, vibrating-microelectrode system. The system enables fluxes to be measured with a spatial resolution of 2-3 microm and a temporal resolution of 10 s. No difference in root porosity was found among the genotypes when grown for 30 days in an aerated solution. Under normoxic conditions, O2 influx was characterised by two distinct peaks, one in the division zone and the other in the elongation zone of the roots. This pattern was found in all three species studied, although the fluxes showed a different magnitude. The peak in the elongation zone coincided with maximum relative elemental growth rates. When the energetics of the cell was disturbed by cyanide, both growth and oxygen O2 influxes ceased at the same time. Under hypoxic conditions, V. riparia plants showed a precise strategy directed toward the maintenance of enough O2 for the respiratory needs of mitosis in the apical meristem of the roots. Thus, whereas in the division zone of V. rupestris and V. vinifera, at bulk O2 concentrations of 0.094 mol x m(-3), the O2 influx was reduced by 70.5 and 38.5%, respectively, for V. riparia no variation in the O2 influx was detected down to bulk O2 concentrations of 0.078 mol x m(-3). Moreover, in accordance with the different tolerances of the plants, the Vitis genotypes were found to differ in their radial O2 loss from the adventitious roots when in an O2-free environment. The results are discussed in terms of possible mechanisms of response to anoxia in Vitis species with different tolerances to flooding. PMID- 11882947 TI - The sensitivity of ABI2 to hydrogen peroxide links the abscisic acid-response regulator to redox signalling. AB - ABI1 and ABI2 are two protein serine/threonine phosphatases of type 2C (EC 3.1.3.16) that act as key regulators in the responses of Arabidopsis thaliana (L.) Heynh. to abscisic acid (ABA). They are involved in the control of ABA mediated seed dormancy, stomatal closure and vegetative growth inhibition. Analysis of the enzymatic properties of ABI2 revealed high sensitivities towards protons and unsaturated fatty acids. Furthermore, the protein phosphatase activity of ABI2 is very sensitive to H2O2, which has recently emerged as a secondary messenger of ABA signalling. Upon H2O2 challenge, ABI2 is rapidly inactivated with an IC50 value of 50 microM in the presence of reduced glutathione. Inhibitor studies with phenylarsine oxide and manipulation of the redox status of ABI2 in vitro indicate that oxidation of critical cysteine residue(s) is responsible for inactivation. The levels of the major cellular thiol compounds cysteine and glutathione in leaves and seedlings of A. thaliana are compatible with a physiological role of H2O2 in regulating ABI2 activity. ABI2 is considered to exert negative regulation on ABA action. Thus, transient inactivation of this protein phosphatase by H2O2 would allow or enhance the ABA dependent signalling process. In conclusion, ABI2 represents a likely target for redox-regulation of a hormonal signalling pathway in higher plants. PMID- 11882948 TI - Characterization of the ZAT1p zinc transporter from Arabidopsis thaliana in microbial model organisms and reconstituted proteoliposomes. AB - The ZAT1p zinc transporter from Arabidopsis thaliana (L.) Heynh. is a member of the cation diffusion facilitator (CDF) protein family. When heterologously expressed in Escherichia coli, ZAT1p bound zinc in a metal blot. Binding of zinc occurred mainly to the hydrophilic amino acid region from H182 to H232. A ZAT1p/ZAT1p*Delta(M1-I25) protein mixture was purified and reconstituted into proteoliposomes. Uptake of zinc into the proteoliposomes did not require a proton gradient across the liposomal membrane. ZAT1p did not transport cobalt, and transported cadmium at only 1% of the zinc transport rate. ZAT1p functioned as an uptake system for 65Zn2+ in two strains of the Gram-negative bacterium Ralstonia metallidurans, which were different in their content of zinc-efflux systems. The ZAT1 gene did not rescue increased zinc sensitivity of a Delta ZRC1single-mutant strain or of a Delta ZRC1 Delta COT1 double-mutant strain of Saccharomyces cerevisiae, but ZAT1 complemented this phenotype in a Delta SpZRC1 mutant strain of Schizosaccharomyces pombe. PMID- 11882949 TI - Questioning the role of salicylic acid and cytosolic acidification in mitogen activated protein kinase activation induced by cryptogein in tobacco cells. AB - Elicitors of plant defence reactions, oligogalacturonides and cryptogein, an elicitin produced by Phytophthora cryptogea, were previously shown to induce a rapid and transient activation of two mitogen-activated protein kinases (MAPKs) in cells of tobacco [ Nicotiana tabacum L. cv. Xanthi; A. Lebrun-Garcia et al. (1998) Plant J 15:773-781]. We verified that these two MAPKs correspond to the salicylic acid-induced protein kinase (SIPK) and the wound-induced protein kinase (WIPK). The involvement of salicylic acid (SA) in cryptogein-induced MAPK activation was investigated using transgenic NahG tobacco cells expressing the salicylate hydroxylase gene and thus unable to accumulate SA. The large and sustained activation of both MAPKs by cryptogein was maintained in transgenic cells compared with non-transgenic tobacco cells. Moreover, weak acids, namely SA, 4-hydroxybenzoic acid, an ineffective analogue of SA in plant resistance, and butyric acid acidified the cytosol, a physiological event also induced by cryptogein, but activated both MAPKs only slightly and transiently in tobacco cells. These results indicate that MAPK activation by cryptogein is not mediated by SA, that cytosolic acidification can be transduced by MAPKs, and that in cryptogein-treated cells, cytosolic acidification should contribute poorly to MAPK activation. PMID- 11882950 TI - Extracellular freezing in leaves of freezing-sensitive species. AB - Low-temperature scanning-electron microscopy was used to study the freezing of leaves of five species that have no resistance to freezing: bean (Phaseolus vulgaris L.), tobacco (Nicotiana tabacum L.), tomato (Lycopersicon esculentum L.), cucumber (Cucumis sativus L.), and corn (Zea mays L.). In the leaves of the four dicotyledonous species, ice was extracellular and the cells of all tissues were collapsed. In contrast, in maize leaves ice was extracellular in the mesophyll, and these cells were collapsed, but the epidermal and bundle-sheath cells apparently retained their original shapes and volume. It is concluded that the leaves of the freezing-sensitive dicotyledonous species tested were killed by cellular dehydration induced by extracellular freezing, and not by intracellular freezing. Freezing injury in maize leaves apparently resulted from a combination of freezing-induced cellular dehydration of some cells and intracellular ice formation in epidermal and bundle-sheath cells. PMID- 11882951 TI - Ethylene response to pollen tube growth in Nicotiana tabacum flowers. AB - In flowers of Nicotiana tabacum L., pollination induces a transient increase in ethylene production by the pistil. The characteristic dynamics of the increase in ethylene correspond to the main steps of the pollen-tube journey into the pistil: penetration into the stigma, growth through the style, entry into the ovary and fertilization. Ethylene is synthesized de novo in the pistil, and its production is reduced in the dark. Ethylene production was monitored in tobacco flowers after pollination with incongruous pollen from three different Nicotiana species, N. rustica, N. repanda and N. trigonophylla, and with pollen from Petunia hybrida. Pollen from all of these different sources can germinate on the stigma surface but each pollen type shows a different behavior and efficiency in penetrating the pistil tissues. Thus, these different crosses provided a model with which to study the response of the pistil to pollination and fertilization. Ethylene evolution upon pollination in tobacco differed in each cross, suggesting that ethylene is correlated with the response to pollen tube growth in the tobacco flower. PMID- 11882952 TI - The sugary-type isoamylase in wheat: tissue distribution and subcellular localisation. AB - To gain an increased understanding of the role of isoamylase (EC 3.2.1.68) in amylopectin synthesis, we studied the tissue-specific distribution and subcellular localisation of this enzyme in wheat (Triticum aestivum L.). A cDNA for wheat isoamylase was isolated from an endosperm-specific library and the missing 5' end was amplified by anchored polymerase chain reaction. Isoamylase transcripts were detected in reproductive and vegetative tissues, with the highest levels occurring in developing kernels. Wheat kernels were then dissected into embryo, endosperm, pericarp and chlorophyll layer, and subjected to protein blot analysis. Isoamylase was most abundant in the endosperm. Within the endosperm, the vast majority of isoamylase was soluble. A much smaller amount of the enzyme was associated with starch granules. Isoamylase was not trapped within starch granules and was absent from dry seeds. Isoamylase was also present in green tissue, which suggests a role in the synthesis of both reserve and leaf starches. PMID- 11882953 TI - Primary care trusts. No can do. AB - Primary care trusts are already chronically short of staff and there is little evidence of workforce planning for the future. Some PCGs, managing sizeable budgets, have no finance staff at all. PCTs will face a shortage of GPs and difficulty retaining nurses. PCTs should make sure they are involved in their local workforce confederation in order to restructure the workforce. PMID- 11882954 TI - Nursing workforce. Skirting the issue. AB - A study of part-time nurses in three London trusts found that they were concentrated in lower grades and less likely to be involved in management than full-time nurses. While senior managers in the trusts were fully committed to flexible working, managers lower in the organisation felt that part-time working presented problems for continuity of care, and that part-time nurses were unwilling to work unsocial hours. A more strategic approach is needed to ensure that part-time nurses are employed most effectively at a time of acute nursing shortages. PMID- 11882955 TI - Primary care trusts. Nurture not nature. AB - Primary care groups had few precedents in the NHS, but their introduction was met with little political hostility. It is difficult to discern PCG/Ts' achievements. The record on commissioning and health improvement is poor. They have not exploited their potential to change the whole NHS. PMID- 11882956 TI - Data briefing. US vs. UK. PMID- 11882957 TI - E-learning. Wire wise. PMID- 11882958 TI - European Computer Driving License. Life in the fast lane. PMID- 11882959 TI - Leadership London. Masters of diplomacy. PMID- 11882960 TI - Workforce development. We can work it out. PMID- 11882961 TI - Overview. Found wanting. PMID- 11882962 TI - Overview/NHS university. Out of the Fryer plan. Interview by Lyn Whitfield. PMID- 11882963 TI - [Clinico-immunological characteristics of lymphoid tumors in children]. AB - The clinical and immunological characteristics of lymphoid tumors were compared in 591 children with acute lymphoblastic leukemia (ALL) and non-Hodgkin's lymphoma (NHL). Comprehensive investigation of a tumor cell by using cytological, morphological, and immunological studies revealed the most significant criteria for differential diagnosis of ALL and NHL in children and showed the specific features of the site of a tumor and the extent of its growth in ALL and NHL in relation to the immunological affiliation of a tumor cell. The predominance of immature forms, such as stem-cell CD34+, pre-pre-B, pre-B and less commonly T cell forms with almost none peripheral B- and T-cell markers could be immunophenotypically detected in ALL. NHL was, on the contrary, characterized by the prevalence of mature immunological subtypes with peripheral B- and T-cell markers and much less frequently pre-B and pre-T cells and at the same time there was no CD34 antigen in the tumor cells. Anaplastic giant lymphoma was a peculiar type of NHL characterized by the presence of large cells having marked anaplasia and expression on the surface of CD30 antigen. A comprehensive study of lymphoid tumors in children showed that immunophenotyping was of great value, whose results were associated with the specific feature of tumor growth and prognosis, which should be borne in mind while planning antitumor therapy programmes. PMID- 11882964 TI - [Growth receptor factor and steroid hormones: their diagnostic value in differential diagnosis of neoplasms in children]. AB - The distribution of epidermal growth factor receptors (EGFR) and receptors for steroid hormones--androgens (RA), estrogens (RE) and glucocorticoids (RG) was studied in malignant (MT) and benign (BT) tumors in children. The distribution and mean levels of some receptors in normal tissues were statistically different from those in MT and BT. The absence of RA and the high levels of RE > 20 fmol/mg protein and RG > 50 fmol/mg protein were typical of MT whereas BT might be characterized by the high levels of EGFR > 300 fmol/mg protein. In MT with and without metastases, the following parameters were significantly different (p < 0.001): RG = 81 and 18 fmol/mg, respectively, and EGFR = 160 and 58 fmol/mg protein, respectively. The most informative parameters for distinguishing BT and MT appeared to be RA (0.086 dit), RE (0.043 dit) and EGFR (0.037 dit). The most sensitive parameter for MT was RG, which, however, is least specific; RA is most specific (75%). The optimal amount of three parameters: RA + RE + EGFR (88.4%) is most exact for differential diagnosis. PMID- 11882965 TI - [Adhesive cell interactions in the biology of cancer]. AB - The present review describes a hypothesis for a critical role of cell adhesive interactions in tumorigenesis. Dysregulation of tissue cell-cell interactions initiates first of all local (in the tissue) and then general (in whole body) conditions for tumor growth. Otherwise imbalance of tissue-specific adhesion factor at the very beginning of carcinogenesis is considered to trigger a cascade of pathological reactions responsible for more severe adhesive disorders that are in turn critical for the "totalitarian" behavior of a tumor and its "colonization" of other tissues and organs. Impaired disturbance is likely to be the key mechanism of carcinogenesis since it is significantly associated with the main features of a tumor: tissue proliferation control loss, anaplasia, invasion, metastasis, and immune surveillance deficit. The hypothesis is supported by evolutionary, biological, histological, immunological, and clinical arguments whose combination does not characterize any other known mechanisms of oncogenesis. The concept of adhesiveness opens new possibilities for the diagnosis, prevention, and treatment of tumors and also improves a strategy for designing new drugs. PMID- 11882966 TI - [Cancer of the proximal section of the stomach: the standards of surgical treatment based on 30 years of experience]. AB - Cancer of the GE-junction is a highly malignant tumor with early lymphatic metastasis to the lymph nodes both in the abdomen and mediastenum. At surgery, lymph nodal metastasis is revealed in nearly 80% of cases. At present, surgical treatment with extended lymph node dissection is the "golden standard". Over 30 year experience in surgically treating GE-junction tumors in 1209 patients at the Thoracoabdominal Department, Russian Cancer Research Center, Russian Academy of Medical Sciences, support the view that extended procedures are superior to standard procedures. Radical procedures were made in 956 of the 1209 patients. These included subtotal proximal gastrectomies in 54.2% of cases, transpleural gastrectomies with esophageal resection in 42.4%, and Ivor-Lewis type procedures in 3.4%. In cases with extended lymph nodal dissection, five-year survival was superior to the results of standard procedures: 32.8 +/- 6.0 and 22.6 +/- 2.8%, respectively (t = 1.8). These figures obviously testify that extended and combined procedures with extended lymph nodal dissection make it possible to stage of a primary tumor and to improve long-term survival. PMID- 11882967 TI - [Problems and achievements in anesthesiology and intensive care in contemporary oncological surgery]. AB - The adoption of aggressive surgery in the Russian Cancer Research Center during recent years as the main treatment philosophy in cancer patients including those at a high and very high risk is very closely connected with appropriate anesthesia and postoperative intensive care. Several organization issues, including technical supply, are discussed in this paper. A concept of anesthesiological support of patients during highly traumatic surgery is presented. The authors share their opinions on infusion therapy, massive blood loss, therapy of perioperative organ and multiple organ failure and sepsis in the light of modern concept of systemic inflammatory response. PMID- 11882968 TI - [The main trends in a complex program of rehabilitation of children with oncological diseases]. AB - The present-day therapeutical programmes allow physicians to cure as many as 70% of children with malignant neoplasms, in some tumor forms, the proportion of recovered children is much higher, which shows it urgent to elaborate comprehensive rehabilitation programmes required for the effective integration of prior patients of children's cancer clinics into a group. The paper presents a complex rehabilitation programme worked out at the Research Institute of Pediatric Oncology and Hematology, Russian Cancer Research Center, Russian Academy of Medical Sciences. The basic directions of the programme are medical, psychological, pedagogical, and social rehabilitation. For medical rehabilitation, phytotherapy, medical laser, climate therapy, and therapeutical exercises are widely used. Of particular value are physical and recreational activities, which include the use of therapeutical exercises, active recreational activities, participation in sporting events, and conditioning swimming. Psychological and pedagogical rehabilitation is aimed at solving personality and familial problems, at developing intellectual abilities, and at eliminating steady-state bad habits and behavioral deviations. A psychological and pedagogical programme is based on the work of art-therapeutical shops where a child realizes his/her strivings, which promotes the recovery of his/her mental and social status. The work of psychologists and psychotherapeutists with children's parents occupies a highly important place since correction of child parent relations serves as a necessary basis for mental recovery in a child. The most important factor of social rehabilitation is an educational programme aimed at acquiring knowledge and at stimulating cognitive activities. To implement the comprehensive programme increases life quality in children who have sustained cancer diseases, improves their social adaptation and thus facilitates integration of these children in society. PMID- 11882969 TI - [The drop in toxicity and the rise in the effectiveness of antineoplastic chemotherapy by correcting the activity of liver monooxygenases: from the experiment to the clinical practice]. AB - The paper reviews both the data available in the literature and the authors' own results of long-term experimental and clinical investigations of the involvement of hepatic monooxygenases (HMO) in the biological activity of antitumor drugs. It reports data of evaluation of HMO activity in pediatric and adult cancer patients, which has shown a decrease in HMO activity in one third of patients without clinical signs of hepatopathy and two thirds of those with toxic hepatic damages after prior chemotherapy. Decreased HMO activity has been found to be stimulated with the enzyme inductor zyxorin. Altered biochemical parameters, such as total bilirubin, ALT and AST, can be corrected with HNO, even if they show a 10-fold deviation from the normal physiological level. The efficacy of zyxorin was tested in patients with advanced cancer and concomitant toxic or viral hepatic disorders (grades II-IV by the WHO classification). Stimulation of inhibited HMO activity allows both decrease and prevention of the manifestations of hepatic toxicity due to anticancer chemotherapy providing a beneficial effect, the dose of cytostatics being not reduced. The authors concluded that the findings provide strong evidence for their assumption that the efficiency of antitumor chemotherapy can be enhanced in patients with concurrent hepatic abnormality by stimulating monooxygenases whose activity is diminished in the majority of these patients. PMID- 11882970 TI - [Contemporary possibilities of clinical immunology in oncology]. AB - Clinical immunology involves studies of immunological diagnosis and monitoring of cancer patients, their immunity and treatment-induced changes, including those associated with immunotherapeutical exposures, immunodiagnosis of leukemias, estimation of solid tumor markers. Immunophenotyping of peripheral blood cells from patients with solid tumors treated with chemo- and immunotherapy has elucidated some relationships to evaluate the efficiency of treatment. Detailed characterization of the structure of leukemic clone has promoted development of individual treatment regimens. The authors' experimental studies will evaluate the effects of cytokines on the mechanisms of multidrug resistance caused by mdr 1 gene hyperexpression and on apoptosis processes diminished by bcl-2 gene hyperexpression. PMID- 11882971 TI - [Pharmacological aspects in the development of liposomal medicinal preparations for the internal injection of hydrophobic cytostatics]. AB - To improve the action and selectivity of new drugs on tumor cells and the use of currently available pharmaceutical technologies to develop the systems of controlled transport of well-known antitumor compounds is one of the ways of enhancing the efficiency of drug therapy for tumors. The tropicity of steroid hormones to definite organs and tissues makes it possible to use them as specific messengers of alkylating groups to target tissues and tumors. Hormone cytostatics synthesized by this principle have a double mechanism of hormonal and cytotoxic actions. The original Russian water-insoluble hormone cytostatistics testifenon, kortifen, and cytestrol acetate demonstrated local tissue irritation together with high antitumor activity. No rational dosage forms make it possible to conduct clinical trials by parenteral administration. The aim of this paper is to summarize the authors' results of designing hydrophobic antitumor hormone cytostatics. The advantages and disadvantages of different approaches to designing traditional dosage forms and to applying colloid liposomal systems for intravenous administration of water-insoluble agents are shown. PMID- 11882972 TI - [The experience in the use of DNA flow cytometry to predict the natural history of malignant neoplasms]. AB - The aim is to study the pattern of a cell population (ploidy) and to evaluate the latter's significance in the prognosis of different carcinomas. A total of 467 patients with primary carcinomas of the breast (n = 177), lung (n = 88), head and neck (n = 140), and colon and rectum (n = 62) were examined. All the patients underwent surgery alone or in combination with radiotherapy (X-ray)/chemotherapy. A follow-up lasted 6 to 120 months. DNA was measured in the operative tumor tissue specimens by using an ICP-22 flow cytometer. Diploid and aneuploid cancers occurred in 20.4-53.6 and 43.6-79.6% of cases, respectively. The frequency of recurrences was mainly associated with aneuploidy and it was more than thrice higher than that with diploidy (21.0-43.2 versus 4.5-14.4%). Five-year survival was twice worse in patients with aneuploid tumors than in those with diploid ones. The survival rates after one operation, preoperative, and postoperative radiation therapy was greater in patients with diploid tumors than in patients with aneuploid ones. Thus, DNA-ploidy of the study neoplasms is of high informative value in predicting the course of a tumorous process and in choosing treatment policy on an individual basis. PMID- 11882973 TI - ["Augmentin" in the antibiotic prevention of postoperative infections complications in gynecological cancer patients]. AB - Hysterectomies are arbitrarily pure operations and thus require antibiotic prevention. A great risk for postoperative infections is particularly high after Wertheim's operation. This study evaluated the efficiency of infection prevention in 19 cancer gynecological patients by using the antibiotic augmentin (amoxicillin/clavulanic acid). The first dose was administered 30 minutes before surgery in order to achieve the maximum drug concentration in blood and tissue at surgery. There was no wound suppuration. One patient (5.3%) undergone Werthein's operation was diagnosed to have retroperitoneal hematoma suppuration. One more patient (5.3%) had fever of unknown genesis. Urinary infections that are not considered to be a sign of inefficiency of antibiotic prevention and that were due to long-term catheter placement developed in 4 (21.1%) of the 19 patients. The postoperative period was smooth in 14 (73.7%) of the 19 patients. The perioperative Augumentin administration regimen used by the authors was easy-to use and beneficial and the drug shows a low toxicity so it may be used for antibiotic prevention in cancer gynecological patients. PMID- 11882974 TI - [Structural and functional features of Epstein-Barr virus LMP-1 gene in patients with anaplastic carcinoma of the nasopharynx in Russia]. AB - Epstein-Barr virus (EBV) is known to be closely associated with the development of anaplastic nasopharyngeal carcinoma (NPC) in some malignancy endemic regions in South-East Asia. LMP1 gene is one of the EBV latent genes, which encodes a latent membrane protein. LMP1 gene is thought to be a classical oncogene since it morphologically transforms cells in vitro and induces tumors in experimental animals in vivo. LMP1 is one of a few genes which is expressed in NPC tissues. It was first shown that C-terminus of LMP1 gene obtained from NPC patients in South East Asia contained a deletion of 30 base pairs (bp). However, this deleted LMP1 gene was then found in the EBV isolates persisting among healthy virus carriers and patients with other EBV-associated abnormalities from both NPC endemic and non-endemic regions. The aim of this investigation was to accomplish a molecular biological analysis of EBV LMP1 genes obtained from Russian NPC patients. To this end, the authors isolated and sequenced the LMP1 clones amplified from the tumor tissues from 7 NPC patients at the N. N. Blokhin Russian Cancer Research Center and primary blood lymphocytes (PBL) from 6 healthy donors. As a result, the authors could not find the deletion of the above-mentioned 30 bp in NPC LMP1 clones, but could in one healthy donor (PBL-2). A functional analysis revealed no significant differences between LMP1 variants with or without 30 bp deletion in their capacity to activate NF kappa B and jun/AP-1 transcription factors. Nevertheless, Russian NPC-derived LMP1 variants as compared with those from PBLs featured some specific amino acid exchanges. These data indicate that the 30 bp deletion of LMP1 gene is not a factor that predisposes to NPC in Russia. PMID- 11882975 TI - [The role of tele-thoracoscopic parasternal lympho-dissection in the diagnosis and treatment of breast cancer]. AB - The outcomes of treatment were studied in 190 patients with breast cancer who had undergone radical mastectomy with telethoracoscopic parasternal lymphodissection. An association of this parameter with the sizes of a primary tumor, with the extent of damage to the axillary lymph collector, with the histological pattern of a tumor, and with the site of a primary focus was analyzed. Radical removal of the parasternal lymphatic chain was proved by scintimammography in the pre- and postoperative periods. The high diagnostic value of telethoracoscopic parasternal lymphodissection was shown as a miniinvasive axilliary of radical mastectomy for breast cancer, which makes it possible to establish a stage of the disease reliably and to plan chemoradiation treatment adequately. PMID- 11882976 TI - [One stage skin-sparing breast repair after mastectomy]. AB - Breast-conserving therapy has become very popular in the past 20 years. This treatment is beneficial only for early stages of breast cancer. When breast conserving therapy is not indicated, skin-sparing mastectomy with immediate breast reconstruction using silicone implants or autogenous tissue is the surgical method of choice. The authors have analyzed the results obtained at three hospitals of Moscow, Novosibirsk, and Tula. A total of 58 breast repairs, including 23 with silicone expanders, 35 with TRAM-flaps were performed by February 2000. They accomplished different designs of skin incisions in relation to the size and site of a tumor and to the shape of the breast. These groups comprised patients with breast cancer T1-2No-1Mo. The follow-up averaged 25.8 months (range 5-52 months). All the patients underwent combined and complex treatments. The median of overall survival was not not achieved. One patient died from dissemination. Two-year acturial survival was 86.8% (one local recurrence). The incidence of postoperative complications was 34.7% in the implant group and 14.2% in the TRAM-flap group. PMID- 11882977 TI - [Contemporary treatment methods in primary breast carcinoma]. AB - Surgery for primary breast carcinoma is the method of choice. The use of the so called conservative (sparing) techniques greatly improves late outcomes. Thus, five-year survival in patients with early cancer stages after radical resections followed by radiation therapy was 87.6% and relapse-free survival was 80.4%. Six courses of adjuvant chemotherapy in 400 patients with Stage IIb provided 5-year overall survival in 81.3% of cases and relapse-free survival in 73.7%. Neoadjuvant chemo- and radiation therapy followed by radical mastectomy preserving both breasts and adjuvant chemo- and hormone therapy increases five year overall and relapse-free survival up to 71.6 and 41.6%, respectively. The promises of improving therapeutical outcomes are associated with early and, in some cases, preclinical diagnosis. PMID- 11882978 TI - [Radiotherapy in multiple modality treatment of children with nephroblastoma]. AB - The role of radiotherapy in multiple modality treatment of Wilms' tumor is evaluated in 225 children aged 3 months to 11.5 years (mean age 3.5 years) with stage III-IV. 184 (81.8%) patients presented with stage III, 93.7% with typical nephroblastoma. Intervention was combined with drug and radiotherapy in 99.6% patients. Exposure of the abdominal cavity in total focal doses of 10.5-50.2 Gy (mean dose 28 Gy) was carried out in 219 (97.3%) of 225 patients. Special attention is paid to the incidence of relapses, remote metastases, and survival of patients in relation to prognostic factors (sex, age, stage of tumor process, terms of exposure, and total focal doses). All patients were followed up for 2 203 months (median 32 months). During this period relapses and/or metastases were observed in 34.2% patients; 30.2% died because of disease progress. 35.1% children live without signs of disease for more than 5 years, 14.7% for more than 10 years. Relapses were more incident during the first year of treatment (in 65% children) and outside the exposed field (72.5% cases). 33.2% patients with stage III developed metastases after 1-49 months; the lungs were involved most often. Prolongation of the period between surgery and exposure of the abdominal cavity led to increase in the incidence of relapses in the abdominal cavity from 6.7% (up to 2 weeks) to 21.9% (more than 1 month), p = 0.02. Relapses were the most frequent in children aged over 4 years. This parameter virtually did not depend on the total focal dose. The absence of relationship between the incidence of local relapses and life span after exposure to a total focal dose of up to 21.6 Gy in comparison with higher doses recommends reduced doses for therapy without notable deterioration of the survival of patients with nephroblastoma. PMID- 11882979 TI - [Osteosarcoma: biochemical and endocrinological studies as a basis for its pathogenetic treatments]. AB - The paper considers data of the long-term studies of androgen metabolism, baseline serum levels of reproductive steroid hormones and their receptors in the tumor, in the blood concentrations of sex steroid-binding globulin and pituitary hormones, the expression of epidermal growth factor, its receptors and ligands, soluble Fas-antigen, vascular endothelial growth factor, angiogenin, and the content of calmodulin, cAMP in the osteosarcoma in 300 patients aged 14 to 56 years, which were made at the N. N. Blokhin Russian Cancer Research Center, Russian Academy of Medical Sciences. Analyzing the findings identified some lines in the study of new pathogenetic treatments for osteosarcoma, aimed at regulating androgen metabolism, correcting the cyclooxygenase pathway of arachidonic acid, the expression of receptors of epidermal growth factor and its ligands, the processes of neoangiogenesis in the tumor. A role of the above parameters in the pathogenesis of osteosarcoma is discussed. PMID- 11882980 TI - [Molecular markers of the cervical cancer]. AB - The genome of human papilloma viruses from a high-risk group (HPV types 16 and 18) has been detected in 90% of cervical tumors and, in some cases, in the adjacent normal tissues. The presence of viral DNA is the main molecular marker of this neoplasia. HPV genome may persist in the tumors as episomal and integrative forms at early and late stages of tumor progression. The status of viral DNA and the pattern of its expression are similar in all cells of this tumor cell population and seem to be a marker of tumor cell monoclonality. Antibodies to the products of viral oncogenes E6 and E7 were found only in 35% of the patients with tumor where HPV genome is present. Thus, this criteria cannot be used for diagnostic and prognostic purposes. On chromosome 6 in the cervical tumors, the specific marker of heterozygocity on loci 6p21.3 was found. The marker appears at the precancer stage and may be regarded as a marker of tumor monoclonality. Heterozygocity loss in the specific locus in the region 6q16-21 correlates with tumor progression and suggests that there are potential tumor suppressor genes in this region of chromosome 6. A group of HPV positive tumors with a hypermethylator phenotype is described. These tumors are characterized by the simultaneous methylation and inactivation of multiple genes, including tumor suppressor genes. PMID- 11882981 TI - Making a mark in 2002. PMID- 11882982 TI - Getting to the point. PMID- 11882983 TI - Storage space is shrinking and inventory is growing--what to do? Spaced out. PMID- 11882986 TI - A standard dilemma. Why the UPN isn't common practice. PMID- 11882987 TI - Stats. Demanding growth. PMID- 11882988 TI - Musculoskeletal ultrasonography in children. PMID- 11882989 TI - Ultrasonography of the shoulder: pitfalls and variants. PMID- 11882990 TI - Sonoanatomy of the median, ulnar and radial nerves. PMID- 11882991 TI - Ultrasonography of lumps and bumps. PMID- 11882992 TI - Ultrasonography of ankle ligaments. PMID- 11882993 TI - Measles virus activates NF-kappa B and STAT transcription factors and production of IFN-alpha/beta and IL-6 in the human lung epithelial cell line A549. AB - Epithelial cells of the respiratory tract are the primary targets of measles virus (MV) infection. In this work we have studied the effect of MV infection on the activation of transcription factors nuclear factor (NF)-kappa B and signal transducer and activator of transcription (STAT) and the production of cytokines in the lung epithelial A549 cell line. NF-kappa B and STAT activation were induced by MV in A549 cells as analyzed by electrophoretic mobility shift assay. NF-kappa B activation was rapid and it was not inhibited by the protein synthesis inhibitor cycloheximide, suggesting that MV directly activates NF-kappa B. In contrast, Stat1, Stat3, and interferon-stimulated gene factor 3 (ISGF3) DNA binding was induced by MV infection with delayed kinetics compared to NF-kappa B activation. MV infection also resulted in an efficient interferon (IFN) alpha/beta and interleukin-6 production. Cycloheximide and neutralizing anti-IFN alpha/beta antibodies inhibited MV-induced activation of Stat1, Stat3, and ISGF3 DNA binding in A549 cells. In conclusion, the results suggest that MV infection activates transcription factors involved in the initiation of innate immune responses in epithelial cells by two different mechanisms: directly by leading to NF-kappa B activation and indirectly via IFN-alpha/beta leading to STAT activation. PMID- 11882994 TI - Different functional domains in the cytoplasmic tail of glycoprotein B are involved in Epstein-Barr virus-induced membrane fusion. AB - A virus-free cell fusion assay relying on the transient transfection of Epstein Barr virus (EBV) glycoproteins into cells provides an efficient and quantitative assay for characterizing the viral requirements necessary for fusion of the viral envelope with the B cell membrane. Extensive cellular fusion occurred when Daudi cells were layered onto Chinese hamster ovary K1 cells transiently expressing EBV glycoproteins gp42, gH, gL, and gB. This is the first direct evidence that gB is involved in the process of EBV entry. Moreover, mutational analysis of gB indicates that the cytoplasmic tail contains two distinct domains that function differentially in the process of fusion. The region from amino acids 802 to 816 is necessary for productive membrane fusion, while amino acids 817 to 841 comprise a domain that negatively regulates membrane fusion. PMID- 11882995 TI - Complexity of the single linear neutralization epitope of the mouse arterivirus lactate dehydrogenase-elevating virus. AB - Results from indirect ELISAs using synthetic peptides of various length that represent segments of the ectodomain of the envelope glycoprotein, VP-3P, of lactate dehydrogenase-elevating virus (LDV) showed that the primary neutralization epitope of LDV is located in a short linear hydrophilic segment in the center of the ectodomain. The epitope becomes slightly altered by amino acid substitutions in the ectodomain and inactivation of virions by various treatments. Neutralizing anti-VP-3P antibodies (Abs) to the epitope interact with the synthetic peptides only if they possess a certain conformation. When the peptides were immobilized on ELISA plates, neutralizing mAbs elicited to inactivated LDV and neutralizing Abs from infected mice bound best to the peptides that consisted of the full-length, 30-amino-acid-long ectodomain. The Abs bound poorly, if at all, to most of the shorter peptides when immobilized, whether truncated at the N- or C-end, but when in solution the same peptides strongly inhibited the binding of the Abs to immobilized full-length peptides. Thus, a conformation of the epitope required for Ab binding and (or) its steric accessibility were lost upon immobilization of the shorter peptides on ELISA plates. Abs raised in mice to peptide-bovine serum albumin conjugates reacted only with immobilized peptides in the indirect ELISA and failed to neutralize LDV. The neutralization epitope of the common LDV quasispecies, LDV-P and LDV-vx, is flanked by N-glycans that block the immunogenicity of the epitope and the neutralization of these LDVs. Abs to a second weakly immunogenic and probably discontinuous epitope appear in LDV infected mice about 1 month postinfection. PMID- 11882996 TI - Val193 and Phe195 of the gamma 1 34.5 protein of herpes simplex virus 1 are required for viral resistance to interferon-alpha/beta. AB - Herpes simplex viruses (HSV) are resistant to the antiviral action of interferon. However, the underlying mechanisms are not well understood. In this report, we show that unlike that of wild-type HSV-1, replication of the gamma 1 34.5 null mutants was significantly inhibited by exogenous interferon-alpha in cells devoid of interferon-alpha/beta genes. Using a series of gamma 1 34.5 deletion mutants, the domain required for interferon resistance was mapped to the region containing amino acids 146 to 263 in the gamma 1 34.5 protein. Interestingly, Val193 Glu and Phe195 Leu substitutions in the protein phosphatase 1 interacting motif of the gamma 1 34.5 protein rendered HSV-1 sensitive to interferon-alpha. Furthermore, gamma 1 34.5 null mutants were sensitive to interferon-alpha/beta in PKR+/+ but not in PKR-/- mouse embryo fibroblasts. These findings provide evidence that the gamma 1 34.5 protein contributes to HSV-1 resistance to interferon-alpha/beta by inhibiting PKR function. PMID- 11882997 TI - Functional expression and membrane fusion tropism of the envelope glycoproteins of Hendra virus. AB - Hendra virus (HeV) is an emerging paramyxovirus first isolated from cases of severe respiratory disease that fatally affected both horses and humans. Understanding the mechanisms of host cell infection and cross-species transmission is an important step in addressing the risk posed by such emerging pathogens. We have initiated studies to characterize the biological properties of the HeV envelope glycoproteins. Recombinant vaccinia viruses encoding the HeV F and G open reading frames were generated and glycoprotein expression was verified by metabolic labeling and detection using specific antisera. Glycoprotein function and cellular tropism were examined with a quantitative assay for HeV mediated membrane fusion. Fusion specificity was verified through specific inhibition by anti-HeV antiserum and a peptide corresponding to one of the alpha helical heptad repeats of F. HeV requires both F and G to mediate fusion. Permissive target cells have been identified, including cell lines derived from cat, bat, horse, human, monkey, mouse, and rabbit. Fusion negative cell types have also been identified. Protease treatments of the target cells abolished fusion activity, suggesting that the virus is employing a cell-surface protein as its receptor. PMID- 11882998 TI - The transmembrane protein of HIV-1 primary isolates modulates cell surface expression of their envelope glycoproteins. AB - We have recently shown that the level of cell surface expression of envelope glycoproteins derived from various human immunodeficiency virus type 1 (HIV-1) primary isolates (PI) was lower than those of envelope glycoproteins derived from T-cell laboratory-adapted (TCLA) HIV-1 (D. Brand et al., 2000, Virology 271, 350 362). We investigated this phenomenon by comparing the cell surface expression of chimeric envelope glycoproteins constructed by swapping the gp120 surface and gp41 transmembrane glycoproteins of the TCLA HIV-1MN and the PI HIV-1(133), HIV 1G365, or HIV-1EFRA. We found that each chimeric envelope construct had a cell surface-specific pattern of expression similar to that of the parental envelope glycoproteins corresponding to the gp41. Thus, the difference in cell surface expression observed between TCLA viruses and various PI is probably due to a signal located in gp41. Identification of this signal may be important for the design of PI envelope-derived immunogens and may increase our understanding of the mechanisms by which HIV-1 escapes from the immune system. PMID- 11882999 TI - The A17L gene product of vaccinia virus is exposed on the surface of IMV. AB - The p21 membrane protein of vaccinia virus (VV), encoded by the A17L gene, has been reported to localize on the inner of the two membranes of the intracellular mature virus (IMV). It has also been shown that p21 acts as a membrane anchor for the externally located fusion protein p14 (A27L gene). Since p14 is located on the surface of IMVs, it is hard to envision that p21 should be located only on the inner membrane. Our results from (i) immunoelectron microscopy, (ii) biotinylation, and (iii) protease treatment of purified IMVs showed that the N terminus of p21 is exposed on the surface of virus particles, while the C terminus is embedded in the membrane. Mono-specific antibodies to the N-terminus of p21 neutralize infection of VV while antibodies to the C-terminal domain do not. We suggest that p21 molecules are located both in the inner and in the outer membrane of IMV. PMID- 11883000 TI - Mutational analysis of early region 4 of bovine adenovirus type 3. AB - The primary objective of characterizing bovine adenovirus type 3 (BAV3) in greater detail is to develop it as a vector for gene therapy and vaccination of humans and animals. A series of BAV3 early region 4 (E4) deletion-mutant viruses, containing deletions in individual E4 open reading frames (Orf) or combinations of Orfs, were generated by transfecting primary fetal bovine retinal cells with E4-modified genomic DNA. Each of these mutants was further analyzed for growth kinetics, viral DNA accumulation, and early-late protein synthesis. Mutant viruses carrying deletions in Orf1, Orf2, Orf3, or Orf4 showed growth characteristics similar to those of the E3-deleted BAV3 (BAV302). DNA accumulation and early/late protein synthesis were also indistinguishable from those of BAV302. However, mutant viruses carrying a deletion in Orf5, Orfs 1-3 (BAV429), or Orfs 3-5 (BAV430) were modestly compromised in their ability to grow in bovine cells and express early/late proteins. E4 mutants containing larger deletions, Orfs 1-3 (BAV429) and Orfs 3-5 (BAV430), were further tested in a cotton rat model. Both mutants replicated as efficiently as BAV3 or BAV302 in the lungs of cotton rats. BAV3-specific IgA and IgG responses were detected in serum and at the mucosal surfaces in cotton rats inoculated with mutant viruses. In vitro and in vivo characterization of these E4 mutants suggests that none of the individual E4 Orfs are essential for viral replication. Moreover, successful deletion of a 1.5-kb fragment in the BAV3 E4 region increased the available insertion capacity of replication-competent BAV3 vector (E3-E4 deleted) to approximately 4.5 kb and that of replication-defective BAV3 vector (E1a-E3-E4 deleted) to approximately 5.0 kb. This is extremely useful for the construction of BAV3 vectors that express multiple genes and/or regulatory elements for gene therapy and vaccination. PMID- 11883001 TI - Exchange of three amino acids in the coat protein results in efficient whitefly transmission of a nontransmissible Abutilon mosaic virus isolate. AB - Geminiviruses are transmitted in a circulative manner by whiteflies, leafhoppers, or treehoppers. The whitefly species Bemisia tabaci (Genn.) is the vector for members of the genus Begomovirus. The closely related bipartite Central American begomoviruses Abutilon mosaic virus (AbMV), Sida golden mosaic virus originating from Costa Rica (SiGMV-CR), and Sida golden mosaic virus originating from Honduras (SiGMV-Hoyv) were used to study transmission by their insect vector. The AbMV isolate is defective in transmission, whereas the two Sida-infecting viruses are readily transmitted by B. tabaci. These three viruses are able to form pseudorecombinant viruses by exchange of genomic components. The pseudorecombinant virus SiGMV-Hoyv A/AbMV B was transmissible, whereas the reciprocal pseudorecombinant virus AbMV A/SiGMV-Hoyv B was not transmitted, indicating that DNA B is not involved in the transmission defect. However, the uptake of the pseudorecombinant virus AbMV A/SiGMV-Hoyv B was much better than AbMV itself, indicating that DNA B or DNA B gene products enhance uptake of viral DNA. Exchange of AbMV coat protein with that of SiGMV-CR resulted in a transmissible chimeric AbMV. Mutagenesis of the AbMV coat protein showed that the exchange of two amino acids, at positions 124 and 149, was sufficient to obtain a whitefly-transmissible AbMV mutant. However, when amino acid 174 was altered in addition to amino acids 124 and 149 AbMV was readily transmitted by B. tabaci. From this we conclude that it is not a concise motif, such as the amino acid triplet, aspartate-alanine-glycine (DAG), involved in aphid transmission of potyviruses, that determines transmissibility of begomoviruses by B. tabaci. Instead it is the composition of the coat protein domain from amino acid 123 to 149, as a minimal transmission domain, with the contribution of amino acids 149 to 174 for efficient transmission. PMID- 11883002 TI - Mutational analysis of the genome-linked protein of cowpea mosaic virus. AB - In this study we have performed a mutational analysis of the cowpea mosaic comovirus (CPMV) genome-linked protein VPg to discern the structural requirements necessary for proper functioning of VPg. Either changing the serine residue linking VPg to RNA at a tyrosine or a threonine or changing the position of the serine from the N-terminal end to position 2 or 3 abolished virus infectivity. Some of the mutations affected the cleavage between the VPg and the 58K ATP binding protein in vitro, which might have contributed to the lethal phenotype. RNA replication of some of the mutants designed to replace VPg with the related cowpea severe mosaic comovirus was completely abolished, whereas replication of others was not affected or only mildly affected, showing that amino acids that are not conserved between the comoviruses can be critical for the function of VPg. The replicative proteins of one of the mutants failed to accumulate in typical cytopathic structures and this might reflect the involvement of VPg in protein-protein interactions with the other replicative proteins. PMID- 11883003 TI - Reassortant analysis of guinea pig virulence of pichinde virus variants. AB - The new world arenavirus Pichinde (PIC) is the basis of an accepted small animal model for human Lassa fever. PIC (Munchique strain) variant P2 is attenuated in guinea pigs, whereas variant P18 is extremely virulent. Previous sequence analysis of the S segments of these two viruses indicated a small number of possible virulence markers in the glycoprotein precursor (GPC) and nucleoprotein (NP) genes. In order to determine the role of these S segment genes in guinea pig virulence in this system, we have generated reassortant viruses. When tested in outbred guinea pigs, the reassortant containing the S segment from the virulent parent P18 (S18L2) caused significantly higher morbidity than the reciprocal reassortant. This increased morbidity was associated with higher viral titers in serum and spleen. However, the S18L2 reassortant was not as fully virulent in this system as the P18 parent, indicating a role for L segment genes in virulence. PMID- 11883004 TI - The Mus cervicolor MuLV isolate M813 is highly fusogenic and induces a T-cell lymphoma associated with large multinucleated cells. AB - M813 is a type-C murine leukemia virus (MuLV) isolated from the Asian rodent Mus cervicolor. We have recently demonstrated that M813 defines a distinct MuLV receptor interference group. Here we show that M813 rapidly induces fusion of MuLV-expressing fibroblasts from "without," with syncytia being observed within 1 h after exposure to virus. Infection of fibroblasts with MuLV from all tested receptor-interference groups imparts susceptibility to M813-induced fusion, provided the cells also express the M813 receptor. Syncytium induction is also observed in vivo; mice infected with M813 develop a peripheral T-cell lymphoma, which is associated with large multinucleated cells of macrophage origin. A recombinant Moloney MuLV/M813 chimeric virus demonstrated that syncytium induction is a function of the Env SU protein. We postulate that the highly fusogenic property of M813 is attributable to either its unique receptor usage or sequences in the proline-rich domain of the Env protein. PMID- 11883005 TI - Characterization of virus-like particles assembled in a recombinant baculovirus system expressing the capsid protein of a fish nodavirus. AB - Betanodaviruses are causative agents of neurological disorders in several species of fish. We cloned and sequenced the RNA2 segment of two grouper viruses isolated from Epinephelus malabaricus (malabaricus grouper nervous necrosis virus, MGNNV) and Epinephelus lanceolatus (dragon grouper nervous necrosis virus, DGNNV). The sequences of the two RNAs were 99% identical and comparison with previously sequenced RNA2 segments of fish nodaviruses striped jack nervous necrosis virus, Atlantic halibut virus, sea bass encephalitis virus, and greasy grouper nervous necrosis virus (GGNNV) revealed that MGNNV and DGNNV were most closely related to GGNNV. No correlation of sequence with geographical habitat was detected. The MGNNV coat protein, the gene product of RNA2, was expressed in Sf21 cells with a recombinant baculovirus system and virus-like particles (VLPs) spontaneously formed. Two types of VLPs were observed: a slower sedimenting particle was RNase sensitive and stain-permeable, while the faster sedimenting particle survived RNase treatment and was not stain-permeable. An image reconstruction of the latter, obtained with electron cryomicroscopy data, revealed a morphology consistent with T = 3 quasi-symmetry but with features significantly different from insect nodavirus structures at the same resolution. This assembly system allows the first biophysical comparisons of fish and insect nodavirus structure, assembly, and stability. PMID- 11883007 TI - Synergistic neutralizing antibody response to a dengue virus type 2 DNA vaccine by incorporation of lysosome-associated membrane protein sequences and use of plasmid expressing GM-CSF. AB - We have previously shown that a dengue virus type 1 DNA vaccine expressing premembrane (prM) and envelope (E) genes was immunogenic in mice and monkeys and that rhesus monkeys vaccinated with this construct were completely to partially protected from virus challenge. In order to improve the immunogenicity of dengue DNA vaccines, we have evaluated the effect of lysosome targeting of antigens and coimmunization with a plasmid expressing GM-CSF on antibody responses. A dengue virus type 2 candidate vaccine containing prM and E genes was constructed in which the transmembrane and cytoplasmic regions of E were replaced by those of the lysosome-associated membrane protein (LAMP). The modified vaccine construct expressed antigen that was colocalized with endogenous LAMP in lysosomal vesicles of transfected cells, whereas the antigen expressed from the unmodified construct was not. It was hypothesized that targeting of antigen to the lysosomal compartment will increase antigen presentation by MHC class II, leading to stronger CD4-mediated immune responses. Mice immunized with the modified construct responded with significantly higher levels of virus neutralizing antibodies compared to those immunized with the unmodified construct. Coimmunization of mice with a plasmid expressing murine GM-CSF enhanced the antibody response obtained with either the unmodified or the modified construct alone. The highest antibody responses were noted when the modified construct was coinjected with plasmid expressing the GM-CSF gene. These results could form the basis for an effective tetravalent dengue virus DNA vaccine. PMID- 11883006 TI - Characterization of neutralization epitopes of simian immunodeficiency virus (SIV) recognized by rhesus monoclonal antibodies derived from monkeys infected with an attenuated SIV strain. AB - A major limitation in the simian immunodeficiency virus (SIV) system has been the lack of reagents with which to identify the antigenic determinants that are responsible for eliciting neutralizing antibody responses in macaques infected with attenuated SIV. Most of our information on SIV neutralization determinants has come from studies with murine monoclonal antibodies (MAbs) produced in response to purified or recombinant SIV envelope proteins or intact SIV-infected cells for relatively short periods of time. While these studies provide some basic information on the potential immunogenic determinants of SIV envelope proteins, it is unclear whether these murine MAbs identify epitopes relevant to antibody responses elicited in monkeys during infection with either wild-type or attenuated SIV strains. To accomplish maximum biological relevance, we developed a reliable method for the production of rhesus monoclonal antibodies. In the present study, we report on the production and characterization of a unique panel of monoclonal antibodies derived from four individual monkeys inoculated with SIV/17E-CL as an attenuated virus strain at a time when protective immunity from pathogenic challenge was evident. Results from these studies identified at least nine binding domains on the surface envelope glycoprotein; these included linear determinants in the V1, V2, cysteine loop (analogous to the V3 loop in human immunodeficiency virus type 1), and C5 regions, as well as conformational epitopes represented by antibodies that bind the C-terminal half of gp120 and those sensitive to defined mutations in the V4 region. More importantly, three groups of antibodies that recognize closely related, conformational epitopes exhibited potent neutralizing activity against the vaccine strain. Identification of the epitopes recognized by these neutralizing antibodies will provide insight into the antigenic determinants responsible for eliciting neutralizing antibodies in vivo that can be used in the design of effective vaccine strategies. PMID- 11883008 TI - Solitary human endogenous retroviruses-K LTRs retain transcriptional activity in vivo, the mode of which is different in different cell types. AB - Solitary long terminal repeats (LTRs) of human endogenous retroviruses (HERVs), tens of thousands of which are spread all over the genome, contain a variety of potential transcription regulatory elements. Information on transcriptional behavior of individual solitary LTRs, however, is limited. We studied the transcriptional activity of several individual HERV-K LTRs in a variety of tissues and cell lines. The RT-PCR technique targeted at specific amplification of the U3 or U5 regions of individual LTRs together with their unique genomic flanks was used to estimate the content of each region in the transcripts. An unequal abundance of the U3 and U5 regions of the transcripts of the same LTR in different cells and tumors was observed. Each LTR is transcribed differently in different cells or tissues, and transcriptional behavior of different LTRs was different in the same cell line or tissue. The transcriptional status of LTRs varies in response to mitogenic and stress factors and in tumor tissues compared to normal counterparts. The LTRs thus seem to be the subjects of specific transcription regulation. The data obtained indicate that an appreciable fraction of the LTRs retained regulatory potential throughout millions of years of evolution and thus may contribute to the overall transcription regulatory network. PMID- 11883009 TI - HTLV-1 cell lines differ in constitutively activated signaling pathways that can be altered by cytokine exposure. AB - Examination of signaling pathways used by HTLV-1-infected rabbit cell lines revealed differences between one, RH/K30, that mediates asymptomatic infection and another, RH/K34, that causes lethal experimental leukemia. Both lines are IL 2 independent; RH/K30 produces IL-4 while RH/K34 produces IL-10. Examination of the Jak/STAT (Janus kinase/signal transducer and activator of transcription) activation of the lines revealed constitutive phosphorylation of Jak1 in both STAT6 phosphorylation, not previously reported for HTLV-1 cells, was observed in RH/K30; STAT1 and STAT3 were phosphorylated in RH/K34. Treatment with cytokines altered the activation of the STAT proteins: IL-2 induced STAT5 phosphorylation in both lines. Supernatant from RH/K34 or IL-10 induced STAT3 phosphorylation in RH/K30 cells. Supernatant from RH/K30 or IL-4 induced STAT6 phosphorylation in RH/K34 cells, which could be reversed with a Jak kinase inhibitor--AG-490. PMID- 11883010 TI - [Severe complications of pregnancy and delivery: the situation in Lorraine based on the European investigation]. AB - The level of maternal mortality appears to be higher in France than in other European countries according to the data collected in the 1995 European survey. We performed a retrospective analysis of severe hemorrhage, pregnancy induced hypertension, and maternal sepsis in 1995 in the Lorraine region and reviewed the management scheme used in each case. There was one maternal death and 223 cases of severe maternal morbidity (110 cases of hemorrhage, 105 cases of pregnancy induced hypertension, 8 cases of maternal sepsis). The frequency of these maternal diseases was an estimated 8 per 1000 births. Ninety percent of the children (90.7%) were living 7 days after birth. Pregnancy after the age of 35 years, obesity, and an intermediate level of vocational training were well documented high risk factors in the Lorraine area. All of the women who developed complications had been followed regularly during their pregnancy. High parity and a scarred uterus were high risk factors for post partum hemorrhage. About 45% (45.5%) of the patients were transferred to an emergency unit for intensive care. Pregnancy-induced hypertension was treated within the normal hospital network, most of the mothers being transferred to a reference center prior to delivery. This retrospective study demonstrates the need for reporting more information on medical records. The data observed improved our knowledge of the prevalence and management of the main causes of direct maternal death in the Lorraine area. It improved our knowledge on the prevalence and management of the main causes of direct maternal death in Lorraine area. PMID- 11883011 TI - [Avoidable anesthetic complications and their prevention]. AB - Anesthesia-related mortality in France is being studied in a survey that began in 1996 using the United Kingdom survey as a model. We describe here the deaths reported in these two surveys and discuss the management strategies involved. These cases point out the risk involved with aspiration syndrome, difficult intubation, cardiac toxicity of local anesthetics, total spinal anesthesia, and respiratory depression. The methodology of the French maternal mortality survey provides data enabling the institution of preventive measures. Specific actions, some of which would require legislation, are proposed to improve maternal safety. PMID- 11883012 TI - [Maternal mortality: avoidable obstetrical complications]. AB - Since 1996, maternal mortality is registered as part of a permanent confidential inquiry in France. The National Committee has studied all cases recorded to assess the cause of death and the avoidable obstetrical complications involved. Recommendations are proposed. In 1996 and 1997, there were 196 maternal deaths in France; 165 could be analyzed. The cause was obstetrical in 123 cases (74%), non obstetrical in 26 (16%), and unidentified in 16 (10%). Ninety-seven direct deaths occurred (78% of the obstetrical mortality cases); 31 cases of hemorrhage including 19 post partum, 20 cases of pregnancy-induced hypertension, 10 cases of eclampsia and 7 of pre-eclampsia, 16 cases of amniotic fluid embolism, 11 cases of thromboembolism and 10 cases of sepsis. The National Committee considered that 54% of these deaths were avoidable: 87% for hemorrhage, 80% for sepsis, and 65% for hypertensive diseases. The deaths due to amniotic fluid embolism were not considered avoidable. This mortality stemmed from substandard care, delayed treatment, missed diagnosis, and professional errors. Clinical recommendations are proposed for post partum hemorrhage, pre-eclampsia and eclampsia, prevention of maternal infection, and thromboembolism prophylaxy. PMID- 11883013 TI - [Epipage study: mortality of very premature infants and state of progress at follow up]. AB - OBJECTIVE: To study perinatal and neonatal mortality of very preterm infants and to assess the association with birth weight and multiple births. METHODS: Infants enrolled in the Epipage study born between 22 and 32 weeks gestational age in 9 French regions in 1997 were included in this study. The main outcome measure was stillbirth and death before discharge from hospital. RESULTS: During the study period, 4397 births and therapeutic abortions meeting the inclusion criteria were recorded (including 16% therapeutic abortions and 18% stillbirths). Survival rate for babies born between 22 and 32 weeks was 67% of all births (stillborn + liveborn) and 85% among livebirths. Survival rose with increasing gestational age: survival (livebirths) was 50% at 25 weeks gestation, 78% at 28 weeks and 97% at 32 weeks. Survival was lower for infants with a birthweight below the 10th percentile and for multiple-pregnancy infants. The different stages of the follow up planned up to 5 years are presented together with the response rate to the postal follow-up questionnaire. CONCLUSION: This cohort provides mortality data on very premature infants during pregnancy, at birth, during hospitalization before discharge. Survival of liveborn infants was stratified by gestational age and was consistent with other geographically based studies of very preterm infants born in the 1990s. PMID- 11883014 TI - [The Epipage cohort in the Lorraine region]. AB - AIMS: The aim of the EPIPAGE survey is to study the future of very premature babies in France. This article describes the characteristics of the EPIPAGE cohort in Lorraine and the results in terms of early in-hospital mortality and morbidity. METHOD: The descriptive analysis concerns preterms born at [22-32] weeks (GP), LBW preterms (less than 1500 g) at > 33 weeks (H), and controls born at 33-34 weeks (T1) and at 39-40 weeks (T2) in Lorraine in 1997. RESULTS: The sample consists of 572 infants: 433 GP, 36 H, 34 T1, and 69 T2. In group GP, 30% are from multiple pregnancies. 140 infants were stillborn (44 MTP, 72 fetal deaths, 17 prepartum deaths). The survival rate of livebirths is 89%. 293 pre terms were admitted to neonatal care units, 18 (6%) had a serious neurological condition (grade III-IV intraventricular haemorrhage, leucomalacia). The average stay was 60 +/- 33.8 days. The follow-up rate at two years is 88%. CONCLUSIONS: The evaluation at 5 years will serve to determine the types of sequelae and family repercussions of the birth of a very preterm infant. PMID- 11883015 TI - [Maternal mortality and severe morbidity in 3 French regions: results of MOMS, a European multicenter investigation]. AB - Maternal mortality rates vary between different European countries. One hypothesis put forward to explain such differences is the potential discrepancy in the incidence of the main obstetrical complications. A European concerted action designed to estimate the incidence of severe post partum hemorrhage (> 1.5 l), PET, and sepsis was carried out in 1995-96 (MOMS-B survey) using standardized definitions and the same questionnaire in all regions. In the 13 regions in Europe involved in the study, including Champagne-Ardenne, Center and Lorraine in France, 1843 cases of obstetrical complication were identified among 182,589 births. The overall mean rate of severe maternal morbidity was 10.1 for 1000 births. This rate was 8.0 for Lorraine, 6.7 for Champagne-Ardenne and 5.5 for Center. The rates of hemorrhage and PET in the United Kingdom, Belgium and Finland were twice the rates in France and Norway. The inverse was observed for sepsis. Such discrepancies between countries, despite the use of standardized definitions, raises several questions. Was the methodology correctly applied? Were threatening situations correctly assessed in France? Was disease severity assessed in the same way in all countries? Further studies would be required to answer these questions. PMID- 11883016 TI - [Perinatal corticotherapy: updates]. AB - The beneficial effects of antenatal corticosteroid treatment are now well established. Likewise, a functional pulmonary improvement has been demonstrated when corticosteroids are used in neonates with chronic lung disease. However, several questions remain to be answered. This review of recent data suggest the need for an updated policy of treatment to improve prognosis. In our population, antenatal maturation has reached a level of 77% below 32 weeks gestation and is associated with a 50% reduction of the risk of severe respiratory distress syndrome. Antenatal steroids have been shown to be beneficial as soon as 23 weeks gestation, but the indication for treatment needs to be carefully evaluated since side effects appear to overcome benefits above 3 repeated courses of treatment. An optimal interval between each course can be set at 10 to 15 days according to the severity of premature labor and gestational age. Since several experimental and clinical studies suggest an increased risk of neurological disability with dexamethasone as compared with betamethasone, it seems consistent to favor the exclusive use of antenatal betamethasone as well as its postnatal choice when indicated. Postnatal use should be restricted to severe chronic lung disease and pulse therapy is now the optimal choice to reduce side effects. PMID- 11883017 TI - [Limits of antenatal management: technical and ethical aspects]. AB - The lower limit for elective delivery depends on technical and ethical issues. Elective delivery is usually accepted after 26 weeks gestation with a 60% survival rate and a 30% handicap rate. Decision making requires close coordination between obstetricians and pediatricians. Sufficient time must be devoted to providing the parents with adequate information. The physician plays a crucial role in decision making, but parent's information and consent are essential. PMID- 11883018 TI - [Extreme prematurity: the limits of neonatal resuscitation]. AB - How far providing neonatal intensive care to extremely low birth weight infants is appropriate is still a highly controversial issue. Decision making when a poor prognosis has been established may be facilitated by consensus based recommendations and rigorous procedures. In the very majority of situations, the provision of intensive care is advocated at birth a priori. A decision of treatment withholding or withdrawal may eventually be made secondarily, in the case major neurological complications, likely to induce severe long term deficits, are evidenced. In any case, an ethical policy focused on each infant's best interest is justified, while the adoption of a systematic, gestational age or birth weight based restriction of access to intensive care may not be acceptable in most countries. Rigorous criteria must be fulfilled for end of life decision making and procedures. Continuous assistance to the patient and to the parents is key determinant. PMID- 11883019 TI - [The gynecology-obstetric system: programmed death of gynecologic surgery?]. PMID- 11883020 TI - [Neonatal bacterial infection by maternal-fetal contamination: for a change in approach? 1. Detection of Streptococcus agalactiae infection: methods and evaluation of results]. AB - Perinatal group B streptococcal infection has been the subject of numerous studies and despite guidelines established during the last decade remains a frequent disease with high mortality. The basic aim of the guidelines is to screen for Streptococcus agalactiae during the antepartum period in order to institute antibiotic therapy during delivery. A critical review of the literature highlights the real impact and adverse effect of these guidelines: difficult application (only two-thirds of all maternity units have a protocol and compliance is only 75%), maternal risks of antibiotic therapy (especially the emergence of resistant Gram negative bacteria), fetal risks (accentuation of neonatal sepsis with resistant strains, retarded neonatal infections, frequent use of antibiotics with a broader spectrum, higher frequency of nosocomial sepsis). PMID- 11883021 TI - [Neonatal bacterial infection by maternal-fetal contamination: for a change in approach? 2. Uncertainties and proposals]. AB - Based on a critical analysis of the literature, it is clear that even though mortality has decreased to 10-15%, the prevalence of neonatal bacterial infections remains dramatically stable. Precise risk factors can be identified in most cases of neonatal infection, but remain uncertain in many others: Streptococcus agalactiae is found in only 40% of the cases of sepsis; Escherichia coli, Haemophilus influenzae, Pneumococcus, and group A Streptococcus strains should also be considered for a real prophylactic strategy; context (prematurity), lack of a consensual attitude for intrapartum strategies; management schemes for asymptomatic neonates. Based on these observations, we make proposals for a realistic attitude for everyday practice based on risk factors, maternal and neonatal bacterial sampling procedures, and modalities for neonatal antibiotic therapy. PMID- 11883022 TI - [Development of the main indicators of perinatal health in metropolitan France between 1995 and 1998. Results of the national perinatal survey]. AB - OBJECTIVE: To study trends in the main indicators of health, medical practice and risk factors. POPULATION AND METHOD: We compared two samples; both included all stillbirths and live births during one week in France: 13,318 in 1995 and 13,718 in 1998. RESULTS: Between 1995 and 1998, there was an increase in maternal age, a development of some characteristics of care (antenatal care by maternity unit staff, HIV screening procedure, antenatal classes) and an increase in the proportion of caesarean sections and epidurals. The proportion of live births before 37 weeks of gestation increased from 5.4% to 6.2%. This trend is explained by an increase in preterm deliveries among twins. Specific questions in the 1998 questionnaire showed that 6.3% of women had received fertility treatment before this pregnancy, 11.1% had an amniocentesis, and 1.1% had an intrauterine transfer. CONCLUSION: This type of survey carried out every three years can show major changes in health, risk factors and medical practices and can yield quick answers to specific questions. PMID- 11883023 TI - [Antepartum perineal massage: review of randomized trials]. AB - OBJECTIVE: To assess the effectiveness of ante partum perineal massage to reduce the number of perineal injuries and episiotomies through a survey of the existing literature. MATERIAL AND METHODS: [corrected] A search both in English and French on randomized clinical trials using the Medline and Cochrane Library databases. The key words: "Perineum", "massage", "perineum injuries", "randomized controlled trial" were selected from the years 1966 to November 2000. RESULTS: Four randomized controlled trials were found. The definition of the selected issues, as well as the included and excluded criteria varied according to the authors. Perineal massages seemed to reduce the occurrence of perineal injuries and episiotomies, mostly among primipara: Labrecque et al. in 1999, noted an OR of 0.56; 95% CI: 0.61-1.31 and at the opposite an increased rate of intact perineum in the massage group (OR = 1.79; 95% CI 1.27-2.52]; and Shipman et al. in 1997 stressed among women of > or = 30 years old an augmentation of intact perineum in the intervention group (OR = 1.93; 95% CI 1.08-3.48), and in the logistic regression taking into account age and birth weight they found a reduction of episiotomies and important perineal injuries (p = 0.02). CONCLUSION: Ante partum perineal massages would seem valid but further studies would be necessary to evaluate the utility of this intervention in the avoidance of serious perineal injuries and the women's satisfaction. PMID- 11883024 TI - [Rectal administration of misoprostol for delivery induced hemorrhage. Preliminary study]. AB - OBJECTIVE: Assess the efficacy of one misoprostol tablet (Cytotec) in severe delivery-induced hemorrhage on uterine atony after failure of syntocinon. PATIENTS AND METHODS: A descriptive study was conducted at the Djibouti Army Hospital from September to December 2000. Five patients with severe delivery induced hemorrhage due to uterine atony unresponsive to syntocinon were give a tablet of misoprostol by rectal administration. RESULTS: The hemorrhage ceased in less than 5 minutes in all patients. No immediate side effect was observed. DISCUSSION: These preliminary findings are encouraging and suggest that further investigations would be useful to assess the beneficial effect of misoprostol by rectal administration for patients with severe delivery-induced hemorrhage due to uterine atony. Case-control studies with a sufficient number of cases should be undertaken to determine the real efficacy of misoprostol in this indication. PMID- 11883025 TI - [Role of misoprostol in the delivery outcome]. AB - OBJECTIVES: Delivery-induced hemorrhage is defined as a blood loss greater than 500 ml within the first 24 hours after delivery. Loss of more than 1000 ml is a sign of gravity. For certain authors, 40% of these hemorrhages could be avoided with systematic preventive measures using uterotonic agents to control the third phase of labor. The aim of our work was to assess the preventive efficacy of active management measures during the third phase of labor and to determine which agents are most effective. PATIENTS AND METHODS: We compared two protocols for controlled deliver: a conventional method using ocytocin (2.5 IU i.v. bolus), and a more recent method using a prostaglandin E1 analog: misoprostol (Cytotec, 3 tablets per os). We compared the two methods with a control group where no preventive measures were used, the standard procedure in our maternity unit. RESULTS: Six hundred two women participated in the study. They were divided into 3 homogeneous groups (ocytocin group misoprostol group, control group). There was a 46% reduction in delivery-induced hemorrhage in the ocytocin group but only a minimal preventive effect against severe hemorrhage. Misoprostol did not demonstrate any efficacy in our study. DISCUSSION: It would appear appropriate to take preventive measures against delivery-induced hemorrhage for all deliveries. A bolus intravenous injection of ocytocin immediately after delivery should bed used. The dose should be greater than that used in this study in order to prevent the development of severe hemorrhage. The most satisfactory results can be obtained with 5 IU (1 ampoule of Syntocinon). It is important to obtain a precise quantification of excessive blood loss in order to institute appropriate care rapidly. Misoprostol should be assessed with other prospective studies because of its easy administration, its low cost and easy storage, important advantages in countries with limited resources. PMID- 11883026 TI - [Hysterectomy for benign lesions: what remains for the abdominal route?]. AB - OBJECTIVE: We conducted a prospective study to assess what indications remain for abdominal hysterectomy for benign conditions. PATIENTS AND METHODS: Four hundred fifty-three hysterectomies were performed for benign conditions between April 4, 1996 and December 14, 1998 in our unit. Abdominal hysterectomy was chosen for large uteri, when vaginal access was unusable, or in case of suspected post operative pelvic adhesions. RESULTS: We performed 330 vaginal hysterectomies (72.85%), 71 laparoscopy-assisted vaginal hysterectomies (15.6%) and 52 abdominal hysterectomies (11.4%). In 10 cases, laparoconversion was required when a vaginal or a laparoscopy-assisted hysterectomy was complicated by pelvic adhesions or poor vaginal access. Mean operative time for abdominal hysterectomy was 130 minutes and mean blood loss was 504 ml. Mean uterus weight was 612 g and mean hospital stay 6.4 days. We had 6 complications (11.53%) (one bladder injury, one ilial injury and four cases of intraoperative hemorrhage). DISCUSSION: Our experience showed that the rate of complications is greater with laparoscopic hysterectomy due to patient selection and should be performed by trained surgeons. PMID- 11883027 TI - [Missed diagnosis of postpartum hemorrhage]. AB - OBJECTIVE: The purpose of this work was to search for the reasons why the diagnosis of post-partum hemorrhage may be missed. PATIENTS AND METHODS: We reviewed retrospectively the files 5,517 vaginal delivery patients cared for in our Gynecology and Obstetrics Unit at the Rouen University Hospital between January 1, 1997 and September 30, 1999. Among these patients, 90 (1.63%) developed anemia during the post-partum period with a 10-point fall in the hematocrit from the pre-delivery level, which corresponds to a 1 liter loss of blood, but for whom no diagnosis of hemorrhage was made. We compared this group with the population of patients who had had a diagnosis of hemorrhage. The chi squared or Fischer's exact text were used where appropriate to compare means and calculate z. RESULTS: Significant risk factors were: primiparity, anemia before delivery, labor induction, locorregional anesthesia, use of ocytocin, long labor with a long active phase, fever during labor, episiotomy and prolonged delay between delivery and onset of suture. CONCLUSIONS: Missed diagnosis appeared to be related to use of visual assessment to determine the degree of bleeding, a very mediocre indicator, particularly when no means of blood collection or quantification is used. In addition, factors having an impact on the development of hemorrhage should be revisited with particular attention given to primiparous patients. PMID- 11883028 TI - [Association of fallopian tube cancer and polymyositis. Apropos of 1 case]. AB - A 51-year-old woman consulted for pelvic pain, metrorrhagia and leukorrhea. Physical examination revealed a renitent and mobile mass in the pelvis. A right lateral uterine mass with hydroxalpinx was found at ultrasonography. Pathology examination of the right annexectomy specimen provided the definitive diagnosis: fallopian tube cancer with polymyositis. No residual tumor was found at total hysterectomy with total bilateral annexectomy. The patient was lost to follow-up for three years without complementary treatment then consulted later for functional disability of the upper then lower limbs with myalgia, swallowing disorders and left supraclavian node enlargement resulting from pelvic relapse of the right fallopian tube adenocarcinoma and left supraclavian metastasis with paraneoplastic polymositis. The patient was given 6 courses of chemotherapy with radiotherapy (45 Gy) centered on the left clavian region. The patient exhibited a spectacular response, and remains in complete remission 50 months after diagnosis. The association of a fallopian tube tumor with polymyositis is exceptional, requiring rapid anticancer treatment effective against the cancer and the paraneoplastic polymyositis. PMID- 11883029 TI - [Apropos of the debate on breech presentation. What is the position does France face in the debate of delivery practices in the case of breech presentation at term?]. PMID- 11883030 TI - [Depression worsens the prognosis of infarct patients. Does psychotherapy prevent coronary death? (interview by Dr. Beate Schumacher)]. PMID- 11883031 TI - [Chronic pancreatitis. Only calcinosis proves the diagnosis!]. PMID- 11883032 TI - [Smoking cessation. A physician's responsibility]. AB - Giving up smoking is a cost-effective measure in the secondary prevention of chronic arterial disease and chronic obstructive pulmonary disease. The involvement of the physician in the primary prevention of smoking and kicking the habit in the case of tobacco-related disease, must receive greater emphasis than has so far been the case in Germany. Weaning smokers suffering from tobacco related disease from their habit is a task for the physician, and may take the form either of a single minimal intervention, or successive consultations that can be integrated in every medical activity. The concept of stepwise smoking dishabituation is supported by evidence-based consensus recommendations on the part of relevant national and international medical societies and public institutions. PMID- 11883033 TI - [Nicotine substitution per inhaler. The "cigarette" against addiction]. PMID- 11883034 TI - [Emergencies in general practice. Flank pain]. PMID- 11883035 TI - [High healing chances in Hodgkin disease, but: now health insurance is turning off the money faucet]. PMID- 11883036 TI - [Anticipating exacerbation. Also consequent treatment for minor allergies]. PMID- 11883037 TI - [Aromatase inhibitors of the 3rd generation. What can the "pill against breast cancer" really do?]. AB - Metastatic cancer of the breast in postmenopausal women can be treated with a number of "hormone-active" substances. The drugs of first choice are still anti estrogens. Today, the three highly selective oral aromatase inhibitors anastrozole, letrozole and exemestane are additionally available for use in continuing progression under anti-estrogen treatment. Roughly one woman in three derives benefit from these new medications as reflected by objective remission or stabilization of the disease for more than 6 months. Neither chemical structure (steroidal/non-steroidal), nor the different nature of inhibition of the active centre of the aromatase, nor whether the inhibition of the enzyme is reversible or irreversible, has any influence on the parameters: response rate, response duration and clinical benefit. PMID- 11883038 TI - [Interview with Professor Dr. Volker Diehl, Cologne. Hodgkin disease: on the path to causal therapy? (interview by Christine Vetter)]. PMID- 11883039 TI - [Pica--the desire for unpalatable substances. Feces, hair and loam as palate teasers? (interview by Dr. med. Julia Rautenstrauch)]. PMID- 11883040 TI - [A new score unmasks individual myocardial infarct risk. From 45 points on one must intervene]. PMID- 11883041 TI - [Patient with skeletal metastases. Mobile despite destroyed bones]. AB - Bone metastases endanger both the mobility of the cancer patient and his ability to profit from medical/nursing care. Treatment aims to prevent fractures of the long bones and vertebrae and to avoid neurological defects caused by compression of the spinal cord. Treatment is indicated when the following criteria are met: for the long bones a defect measuring more than 2.5 cm in length or affecting more than 50% of the circumference; for the vertebrae destruction of more than 60%. Almost all patients require radiotherapy, with a minority also needing surgical treatment in the form of intralesional tumor resection, closure of the defect with bone cement, and the use of metal implants. PMID- 11883042 TI - [Liver metastases in colorectal carcinoma. With complete resection healing is possible]. AB - Surgical treatment of liver metastases from colorectal carcinoma can achieve 5 year survival rates of up to 40%, and 10 year survival rates of up to 20%, making resection the only potentially curative treatment option. This applies, however, only when metastatic disease is confined to the liver and when complete removal of the metastases is possible. Patients with liver metastases from non-colorectal primaries--with the exception of the rare neuroendocrine tumors--are not usually candidates for surgery. PMID- 11883043 TI - [Brain metastases. Possibilities and limits of treatment]. AB - The objective of the treatment of brain metastases is not merely to control the disease, but also to preserve an acceptable quality of life by keeping neurological symptoms in abeyance for as long as possible. In most cases, symptoms may be due to considerable perifocal edema. This is responsive to treatment with steroids, and these are therefore given to all patients. Specific treatment regimens include microsurgical removal of metastatic lesions, radiosurgery, irradiation of the brain, and chemotherapy. The choice of the treatment modality is dictated by the general state of health of the patient, the location and size of the metastases, the number of such lesions, and systemic involvement. PMID- 11883044 TI - [Removing lung metastases. With surgical success 36% of patients survive 5 years]. AB - Resection of lung metastases can improve the long-term survival of many oncological patients. This can be achieved only by complete resection, which necessitates careful prior staging. With few exceptions (renal cell carcinoma, carcinoma of the colon metastasizing to the lungs and liver, chest wall and lung metastases) no extrapulmonary metastases should be present. Solitary round lesions should always be resected, since the prognosis for single lesions is favorable, and the primary may be a bronchial carcinoma, which is often curable. Although overall morbidity and mortality rates are low, the indication for surgical treatment of lung metastases should nevertheless be considered with care. PMID- 11883045 TI - [From neuromuscular blocker to analgesic. What is the value of botulinum toxin in pain therapy?]. PMID- 11883046 TI - [Cardiac arrhythmias in advanced age. That could be a thyrotoxic crisis!]. PMID- 11883047 TI - [Phytopharmaceuticals in general practice. Proven aids of the practitioner]. PMID- 11883048 TI - [Are you fit for carnival? Prescriptions to control breath alcohol and hangover]. PMID- 11883049 TI - [Oral contraceptives. 2: Side-effects and interactions]. PMID- 11883050 TI - [Despite unremarkable diagnostic technique thus far, dyspnea at night. Unstable angina pectoris]. PMID- 11883051 TI - [Drug interactions caused by phytopharmaceuticals. Don't underestimate herbs!]. PMID- 11883052 TI - Don't miss smallpox/plague outbreaks: adapt strategies to track bioterrorism. AB - Your strategy for tracking bioterrorism should focus on early recognition of unusual disease patterns, prompt notification of public health agencies, and informing staff about additional precautions to take if an outbreak is suspected. Instead of general awareness training, have a local expert address symptoms of a specific agent, such as smallpox. Post information about tracking patterns of illness in all clinical areas. Ensure that after-hours cases aren't overlooked by having staff report suspicions by computer or telephone. PMID- 11883053 TI - Cutting-edge system spots outbreaks before you do. PMID- 11883054 TI - Here's what new ED ultrasound guidelines say. AB - New ED ultrasound guidelines from the American College of Emergency Physicians give recommendations for training, scope of practice, quality assurance, and certification. The guidelines recommend that 150 ultrasound examinations be completed, but if an individual isn't proficient, more examinations must be required. All ultrasound studies during the credentialing period should be compared to gold standard studies, such as computed tomography scan or operative reports. You must be clear about the limitations of ED ultrasound examinations when consulting with specialists. PMID- 11883055 TI - Does EMTALA apply during a disaster? AB - A statement from Medicare says that disaster plans for transferring triaged patients may supercede the usual EMTALA requirements, but this is true only in certain scenarios. During a disaster, it's unacceptable to transfer a patient to a more appropriate facility if doing so amounts to refusal of lifesaving care. Don't assume that patients will be diverted successfully to appropriate facilities after a large-scale disaster. The required level of documentation under EMTALA may not be possible during a disaster, but some type of documentation is necessary. PMID- 11883057 TI - Patient information sheet: anthrax. PMID- 11883056 TI - Distinguishing smallpox from chickenpox. PMID- 11883059 TI - Bioterrorism watch. Ring of fire: CDC plan to immunize around first smallpox cases has the devil in the details. PMID- 11883060 TI - Distinguishing influenza-like illness from inhalational anthrax. PMID- 11883061 TI - Bioterrorism watch. Smallpox or chickenpox? How to make the diagnosis. PMID- 11883062 TI - Bioplex technology: novel synthetic gene delivery system based on peptides anchored to nucleic acids. PMID- 11883063 TI - Surgical procedures for intravascular delivery of plasmid DNA to organs. PMID- 11883064 TI - Direct gene transfer into mouse heart. PMID- 11883065 TI - Oligonucleotide-mediated site-directed gene repair. PMID- 11883066 TI - Myoblast-mediated gene transfer for therapeutic angiogenesis. PMID- 11883067 TI - Use of phage display to identify novel peptides for targeted gene therapy. AB - The field of gene therapy has developed at an astonishing pace over the past decade, perhaps too quickly. Clinical studies have highlighted major flaws in the ability of current vectors to deliver genes safely and effectively to patients; hence the further development of vectors is a prerequisite for future success. In this chapter we have discussed advances in development of targeted vectors through isolation of targeting moieties using phage display. The field of gene therapy will benefit considerably by the isolation and use of peptides that are effective for targeting in vivo, particularly for diseases affecting individual organs. Only when truly selective and highly efficient vectors are constructed will the tremendous potential of gene therapy be realized. PMID- 11883068 TI - Helper-dependent adenoviral vectors. PMID- 11883069 TI - Gene transfer methods for transplantation. PMID- 11883070 TI - Generation and growth of gutted adenoviral vectors. PMID- 11883071 TI - Adenovirus-mediated gene transfer for cardiovascular and renal diseases. PMID- 11883072 TI - Gene transfer to blood vessels using adenoviral vectors. PMID- 11883073 TI - Rearrangements in adenoviral genomes mediated by inverted repeats. PMID- 11883074 TI - High-capacity, helper-dependent, "gutless" adenoviral vectors for gene transfer into brain. PMID- 11883075 TI - Gene therapy for hypertension: the preclinical data. AB - In spite of several drugs for the treatment of hypertension, there are many patients with poorly controlled high blood pressure. This is partly due to the fact that all available drugs are short-lasting (24 hr or less), have side effects, and are not highly specific. Gene therapy offers the possibility of producing longer-lasting effects with precise specificity from the genetic design. Preclinical studies on gene therapy for hypertension have taken two approaches. Chao et al. have carried out extensive studies on gene transfer to increase vasodilator proteins. They have transferred kallikrein, atrial natriuretic peptide, adrenomedullin, and endothelin nitric oxide synthase into different rat models. Their results show that blood pressure can be lowered for 3 12 weeks with the expression of these genes. The antisense approach, which we began by targeting angiotensinogen and the angiotensin type 1 receptor, has now been tested independently by several different groups in multiple models of hypertension. Other genes targeted include the beta 1-adrenoceptor, TRH, angiotensin gene activating elements, carboxypeptidase Y, c-fos, and CYP4A1. There have been two methods of delivery antisense; one is short oligodeoxynucleotides, and the other is full-length DNA in viral vectors. All the studies show a decrease in blood pressure lasting several days to weeks or months. Oligonucleotides are safe and nontoxic. The adeno-associated virus delivery antisense to AT1 receptors is systemic and in adult rodents decreases hypertension for up to 6 months. We conclude that there is sufficient preclinical data to give serious consideration to Phase I trials for testing the antisense ODNs, first and later the AAV. PMID- 11883076 TI - Gene therapy methods in cardiovascular diseases. AB - Local gene transfer into the vascular wall offers a promising alternative to treat atherosclerosis-related diseases. Blood vessels are among the easiest targets for gene therapy because of percutaneous, catheter-based treatment methods. On the other hand, gene transfer to the artery wall can also be accomplished from adventitia either by ex vivo gene transfer and implantation of transfected cells or by direct in vivo gene transfer methods. In the future, as the pathological processes in arteries are better understood, several therapeutic genes could be combined and these "gene cocktails" are expected to produce enhanced therapeutic effects in vascular gene therapy. We have developed a new, efficient technique for performing ex vivo gene transfer to rabbit arterial wall using autologous SMC. The cells were harvested from rabbit ear artery, transfected in vitro with VSV-G pseudotyped lacZ retrovirus, and returned back to the adventitial surface of the carotid artery using a silicone collar or collagen sheet placed around the artery. The transduced SMCs implanted with a high efficiency and expressed beta-galactosidase marker gene at a very high level 7 days and 14 days after the operation. The level of lacZ expression decreased thereafter, but was still easily detectable for at least 6 months and was exclusively localized to the site of cell implantation inside the collar. Development of new vectors, such as baculovirus, for gene transfer will provide targeted, efficient, and safer methods for gene delivery. Plasmids and viruses coding for more than one protein, and bearing regulatory elements, would be useful for future gene therapy applications. Also, constructing second-generation viruses that contain fewer endogenous genes in their genome may reduce immunological reactions caused by the first-generation adenoviruses. In conditions where stable expression of therapeutic proteins is needed, it is necessary to develop better ex vivo and in vivo gene transfer strategies. Also, production of viruses that can efficiently transfect nondividing cells will be important for future applications of vascular gene therapy. However, current knowledge from vascular gene transfer experiments strongly suggests that vascular gene transfer is a promising new alternative for the treatment of cardiovascular diseases. PMID- 11883077 TI - Gene delivery to cardiac muscle. PMID- 11883078 TI - Recombinant AAV-mediated gene delivery using dual vector heterodimerization. PMID- 11883079 TI - Designing and characterizing hammerhead ribozymes for use in AAV vector-mediated retinal gene therapies. PMID- 11883080 TI - Stabilized plasmid-lipid particles: a systemic gene therapy vector. AB - The ability of a systemically administered gene therapy vector to exhibit extended circulation lifetimes, accumulate at a distal tumor site, and enable transgene expression is unique to SPLP. The flexibility and low toxicity of SPLP as a platform technology for systemic gene therapy allows for further optimization of tumor transfection properties following systemic administration. For example, the PEG coating of SPLP is necessary to engender the long circulation lifetimes required to achieve tumor delivery. However, PEG coatings have also been shown to inhibit cell association and uptake required for transfection. The dissociation rate of the PEG coating from SPLP can be modulated by varying the acyl chain length of the ceramide anchor, suggesting the possibility of developing PEG-Cer molecules that remain associated with SPLP long enough to promote tumor delivery, but which dissociate quickly enough to allow transfection. Alternatively, improvements may be expected from inclusion of cell specific targeting ligands in SPLP to promote cell association and uptake. Finally, the nontoxic properties of SPLP allow the possibility of higher doses. A dose of 100 micrograms plasmid DNA per mouse corresponds to a dose of approximately 5 mg plasmid DNA per kg body weight. This compares well to small molecules used for cancer therapy, which typically are used at dose levels of 10 to 50 mg per kg body weight. In summary, SPLP consist of plasmid encapsulated in a lipid vesicle that, in contrast to naked plasmid or complexes, exhibit extended circulation lifetimes following intravenous injection, resulting in accumulation and transgene expression at a distal tumor site in a murine model. The pharmacokinetics, biodistribution, and tumor transfection properties of SPLP are highly sensitive to the nature of the ceramide anchor employed to attach the PEG to the SPLP surface. The SPLP-CerC20 system in which the PEG-Cer does not readily dissociate exhibits good serum stability, long circulation lifetimes, and high levels of tumor accumulation and mediates marker gene expression at the tumor site. The flexibility of the SPLP system offers the potential of further optimization to achieve therapeutically effective levels of gene transfer and clearly has considerable potential as a nontoxic systemic gene therapy vehicle with general applicability. These features of SPLP contrast favorably with previous plasmid encapsulation procedures. Plasmid DNA has been encapsulated by a variety of methods, including reverse phase evaporation, ether injection, detergent dialysis in the absence of PEG stabilization, lipid hydration and dehydration-rehydration techniques, and sonication, among others. The characteristics of these protocols are summarized in Table I. None of these procedures yields small, serum-stable particles at high plasmid concentrations and plasmid-to-lipid ratios in combination with high plasmid-encapsulation efficiencies. Trapping efficiencies comparable with the SPLP procedure can be achieved employing methods relying on sonication. However, sonication is a harsh technique that can shear nucleic acids. Size ranges of 100 mm diameter or less can be achieved by reverse-phase techniques; however, this requires an extrusion step through filters with 100 nm or smaller pore size which can often lead to significant loss of plasmid. Finally, it may be noted that the plasmid DNA-to lipid ratios that can be achieved for SPLP are significantly higher than those achievable by any other encapsulation procedure. PMID- 11883081 TI - Adeno-associated viral vector-mediated gene therapy of ischemia-induced neuronal death. PMID- 11883082 TI - Recombinant adeno-associated viral vector production using stable packaging and producer cell lines. PMID- 11883083 TI - Streamlined large-scale production of recombinant adeno-associated virus (rAAV) vectors. PMID- 11883084 TI - Gene transfer to the brain using feline immunodeficiency virus-based lentivirus vectors. PMID- 11883085 TI - Generation of HIV-1 derived lentiviral vectors. PMID- 11883086 TI - Design, production, safety, evaluation, and clinical applications of nonprimate lentiviral vectors. PMID- 11883087 TI - Gene transfer to airway epithelia using feline immunodeficiency virus-based lentivirus vectors. PMID- 11883088 TI - Transduction of a gene expression cassette using advanced generation lentiviral vectors. PMID- 11883089 TI - Construction, purification and characterization of adenovirus vectors expressing apoptosis-inducing transgenes. PMID- 11883090 TI - Ligand-inducible transgene regulation for gene therapy. AB - A synthetic ligand regulable system for gene transfer and expression has been developed in our laboratory based on mechanistic studies of steriod hormone receptor and transcriptional regulation. This gene switch system possesses most of the important features that are required for application of the system in biological research and clinical gene therapy in the future. As the primary ligand tested in this system, mifepristone can effectively turn on the regulatory circuit at doses much lower than those used in the clinic. By modification of the chimeric regulator and its feedback regulatory mode, this system has been optimized to produce very low basal activity with high inducibility in the presence of mifepristone. Also, improvements in regulator composition have been made to minimize immunogenicity and make the system more amenable to human gene therapy. Moreover, incorporation of this gene switch system into the HC-Ad vector system has further enhanced the efficiency of gene transfer and the long-term inducible expression of transgenes. However, for each application within a different biological system, the gene switch needs to be optimized to achieve appropriate inductions. In particular, the method used to deliver the transgenes and adjustment of ligand dosage are critical for in vivo gene expression. PMID- 11883091 TI - Large-scale production of retroviral vectors for systemic gene delivery. PMID- 11883092 TI - Oncoretroviral and lentiviral vector-mediated gene therapy. AB - Oncoretroviral vectors and lentiviral vectors offer the potential for long-term gene expression by virtue of their stable chromosomal integration and lack of viral gene expression. Consequently, their integration allows passage of the transgene to all progeny cells, which makes them particularly suitable for stem cell transduction. However, a disadvantage of oncoretroviral vectors based on Moloney murine leukemia virus (MoMLV) is that cell division is required for transduction and integration, thereby limiting oncoretroviral-mediated gene therapy to actively dividing target cells. In contrast, lentiviral vectors can transduce both dividing and nondividing cells. Lentiviral vectors have been derived from either human or primate lentiviruses, with the human immunodeficiency virus (HIV) as prototype, or from nonprimate lentiviruses, such as the equine infectious anemia virus (EIAV). The ability to pseudotype oncoretroviral and lentiviral vectors with the vesicular stomatitis virus G glycoprotein (VSV-G) allowed for the production of high-titer vectors (10(9) 10(10) transducing units/ml). These high-titer vector preparations were shown to effectively cure genetic diseases in experimental animal models and constitute an essential step toward direct in vivo gene therapy applications. This chapter focuses on different methods that permit large-scale production of high-titer VSV G pseudotyped oncoretroviral and primate or nonprimate lentiviral vectors and highlights their importance for achieving therapeutic effects in preclinical animal models. PMID- 11883093 TI - HSV-1 amplicon vectors. PMID- 11883094 TI - Microencapsulation of genetically engineered cells for cancer therapy. PMID- 11883095 TI - HVJ (hemagglutinating virus of Japan; Sendai virus)-liposome method. PMID- 11883096 TI - Gene transfer with foamy virus vectors. PMID- 11883097 TI - Infectious Epstein-Barr virus vectors for episomal gene therapy. PMID- 11883098 TI - Enhancing direct in vivo transfection with nuclease inhibitors and pulsed electrical fields. PMID- 11883099 TI - Hydrodynamics-based transfection: simple and efficient method for introducing and expressing transgenes in animals by intravenous injection of DNA. PMID- 11883100 TI - Differential screening technology in the service of ovarian biology. PMID- 11883101 TI - The zona pellucida in folliculogenesis, fertilization and early development. PMID- 11883102 TI - GDF-9 and BMP-15: oocyte organizers. PMID- 11883104 TI - The ovarian androgen-producing cells: a 2001 perspective. PMID- 11883105 TI - The ovarian life cycle: a contemporary view. PMID- 11883103 TI - Multiple signal transduction pathways regulate ovarian steroidogenesis. PMID- 11883106 TI - The ovarian gonadotropin receptors in health and disease. PMID- 11883108 TI - [Comparison of invasive diagnostic techniques and revascularization therapy of acute coronary syndrome in Hungarian and international databases]. AB - INTRODUCTION, AIMS, METHODS: The databases of large multicentric trials dealing with acute coronary syndrome (OASIS Registry, CORE, PURSUIT, FRAX.I.S.) were analysed. RESULTS: Significant differences were disclosed comparing the baseline and history characteristics of patients randomized from Eastern or Western Europe. In most investigations the patients from Eastern Europe had a higher prevalence of hypertension, diabetes, angina and myocardial infarction. Invasive diagnostics and revascularization therapy were less frequently used in Eastern than in Western Europe in all the trials. The prevalence of myocardial infarction and the case fatality rate were not uniformly different during the early days, but after 30 days the case fatality rate in Eastern Europe was significantly higher than in Western Europe. CONCLUSION: It is suggested that the differences in demographic and baseline characteristics were only partially responsible for the difference in mortality. The low intervention rate and delayed revascularization may contribute to the high rate of myocardial infarction and mortality in Eastern Europe. PMID- 11883107 TI - Local role of progesterone in the ovary during the periovulatory interval. PMID- 11883109 TI - [Gene therapy in liver diseases]. AB - The basic principles of gene therapy, ex vivo and in vivo gene transfers and various vectors have been reviewed first in the paper. Then the models for clinical application are shown, including the recent results of experimental studies, with special regard to the treatment of liver diseases and amongst them hepatocellular carcinoma. Gene therapy is still in preclinical stage. Due to the ongoing intensive genetic studies and if the existing problems being resolved in this field, we can predict that the early years of this millennium gene therapy will be a useful modality in the clinical hepatology as well. PMID- 11883110 TI - [Human immunodeficiency virus-positive cases among intravenous drug users]. AB - AIM: The authors show human immunodeficiency virus positive cases among Hungarian intravenous drug users. PATIENTS/METHOD: The cases were identified by self admittance (by the results of HIV screening), and by objective methods. The persons were studied by questionnaire on their risk behaviours (injecting and sexual behaviours). Treatment seeking (76 cases) and out-of-treatment (121 cases) groups were analysed. In the first group the self-admittance human immunodeficiency virus-status was taken into consideration, in the second group the objective human immunodeficiency virus-status was studied. RESULTS: In the treatment-seeking group three persons knew that they had been human immunodeficiency virus positive or had had Acquired Immunodeficiency Syndrome related disease. The three males were 25 year-old, homeless, and used heroine intravenously. In the out-of-treatment group one person was identified by objective test, who did not know his infection. This 20 year-old male used heroine and amphetamine intravenously, and his risk behaviour was not characteristic for intravenous drug users. CONCLUSION: In these cases the sexual transmission cannot be excluded, but it is pointed out, that the human immunodeficiency virus is present among intravenous drug users in Hungary. Taking into consideration the risk behaviours, these persons can transmit the disease through injecting and or sexual behaviours. In case of treatment seeking group, the treatment institutions (where the interviews took place) did not know the infections among their patients. According to their results, the authors call the attention to the human immunodeficiency virus infection among Hungarian intravenous drug users and they urge specific interventions for intravenous drug users. PMID- 11883111 TI - [Mucolipidosis II with unusual biochemical parameters]. AB - INTRODUCTION: A clinical diagnosis of I-cell disease was established in an infant with coarse face, hyperplastic gums, severe growth deficiency, skeletal deformities and retarded psychomotor development. PATIENT/METHOD: Albeit, enzyme measurements in our patient leukocytes and serum including arylsulfatase, hexosaminidase, alpha-fucosidase, iduronate-sulphatase and beta-galactosidase showed no significant alteration, in serum an increased activity of beta glucuronidase was detected. Enzyme studies in fibroblasts showed an abnormal intracellular/extracellular distribution of enzyme activities, and significantly decreased activity of UDP-N-acetylglucosamine-1-phosphotransferase confirming the diagnosis of I-cell disease. RESULTS/DISCUSSION: In the mother's next pregnancy at 15th week of gestation lysosomal enzyme activities were determined in amniotic fluid and cultured amniotic cells, and the diagnosis of an unaffected fetus was established. Based on our experience we suggest the use of cell cultures for the diagnostics of this rare disease. PMID- 11883112 TI - [Diffuse splenic metastasis of occult breast cancer with incompatible blood group antigenic determinants]. AB - INTRODUCTION: Cancer cells with immunogenic properties having modified blood group substances are widely studied (Kannagi, 1988, Hakomori, 1999). CASE REPORT: A 78-year-old female patient was admitted to the hospital in terminal state with unsusceptible circulatory failure. At autopsy the spleen (weight: 420 g) was extremely firm with diffuse blackberried colored cut surface. There were no signs of carcinomatous process at autopsy. MATERIAL/METHOD: By histology the spleen showed diffuse metastatic carcinomatous infiltration. Antibody to Breast carcinoma antigen (BioGenex) labelled metastatic cells of the spleen and bone marrow. The patient had O blood group according to her blood group phenotype. The authors used lectins with and without blood group antigen (BGA) specificity and monoclonal antibodies, too. RESULTS: They found that the B blood group specific Bandeiraea simplicifolia agglutinin I lectin and the anti-B mab were labelled intensively all the metastatic cells of spleen and bone marrow. The A BGAs (with anti-A mab and Dolichos biflorus agglutinin) were completely negative. CONCLUSIONS: These observations raise the possibility that the detected incompatible B blood group antigen determinants on the metastatic cells were immunogenic. The authors propose, that the survived carcinoma cells found place of refuge from the immune surveillance in the spleen and in the bone marrow, where the complement mediated tumor cell lysis, immune-rejection was not effective. PMID- 11883113 TI - [Hungarian physicians in Vienna]. PMID- 11883114 TI - [Disorders of blood pressure regulation in the aging population. 1926]. PMID- 11883115 TI - Experimental investigations of image quality in X-ray mammography with conventional screen film system (SFS), digital phosphor storage plate in/without magnification technique (CR) and digital CCD-technique (CCD). AB - Comparison of image quality in X-ray mammography between conventional film screen film system (SFS), digital phosphor storage plate in and without magnification technique (CR) and digital CCD-technique (CCD). Radiograms of an RMI-mammography phantom were acquired using a conventional screen film system, three digital storage plate systems and two digital systems in CCD-technique. Additionally, the radiograms of one digital phosphor storage plate system were post-processed regarding contrast and included in the comparison. The detectability of details was best with the digital mammography in CCD-technique. After confirming these promising results in clinical studies, digital mammography should be able to replace conventional screen film technique. PMID- 11883116 TI - [Multi-slice spiral CT of the coronary arteries: clear vascular imaging using standard software]. AB - OBJECTIVE: To visualize the coronary arteries with a clear view and over a long distance by using data sets from contrast-enhanced computed tomography of the heart. MATERIAL AND METHODS: Image data of 151 patients suffering from coronary artery disease were calculated by means of retrospective triggering at four different diastolic delay times in contrast-enhanced CT. The large coronary segments were subsequently reconstructed in two planes with multiplanar volume reconstruction (MPVR)--a non-dedicated postprocessing software. RESULTS: On the pre-condition that data sets were acquired at sinus rhythm and at a heart beat rate lower than 65/min coronary arteries could be depicted over a long distance in single or double angulated reconstruction planes with the help of multiplanar volume reconstruction (MPVR). Time consumption for image reconstruction was reasonable. Additionally to the anatomy of the coronary arteries in two different planes, typical CT findings in occluding coronary artery disease are presented. CONCLUSION: Multiplanar volume reconstruction (MPVR) implemented on most workstations is a powerful and ideal postprocessing tool in reconstructing coronary arteries from contrast-enhanced CT data sets. PMID- 11883118 TI - [Involvement of the facial skull in Erdheim-Chester disease]. AB - We report on a patient suffering from Erdheim-Chester-disease (ECD). ECD represents a very rare entity with lipogranulomatosis of mesenchymal origin. The most common radiological manifestation is the involvement of the long bones of the extremities. Here we find sclerosis of the spongiosa combined with a thinning of cortical structures. This often results in a small crack of hyperlucency between corticals and spongiosa. Our case demonstrates an involvement of the craniofacial part of the skull showing sclerosis of the upper jaw bone. This manifestation has not yet been reported in the literature. PMID- 11883117 TI - [Multi-slice spiral CT: 3D CT angiography for evaluating therapeutically relevant stenosis in peripheral arterial occlusive disease]. AB - PURPOSE: Evaluation of 3D multislice CT angiography for the assessment of relevant stenoses of pelvic arteries and arteries of the lower extremity in patients with peripheral artery occlusive disease compared to digital subtraction angiography. METHOD/MATERIALS: For this study we examined 31 patients with peripheral artery occlusive disease. All patients received a multislice helical CT angiography and arterial digital subtraction angiography. Multislice CT angiography was performed with a Somatom Plus 4 Volume Zoom (Siemens, Erlangen, Germany). After test bolus injection of 20 ml Ultravist 370 (Schering AG, Berlin) additional 150 ml were applied with a flow rate of 3 ml/sec and a scan delay between 20-35 sec depending on individual blood circulation time. Collimation was 4 x 2.5 mm with a pitch of 6. Reconstructed slice thickness was 3 mm. 3D reconstructions of arteries of pelvic and lower extremity arteries were performed in volume rendering technique on a 3D Virtuoso workstation (Siemens, Erlangen). RESULTS: For the assessment of therapeutically relevant stenoses (over 50% reduction of luminal diameter) multislice CT achieved the following results compared to conventional angiography for the diagnosis of stenosis: sensitivity of 86%, specificity of 86% and an accuracy of 72%. CONCLUSIONS: Multislice helical CT angiography of pelvic arteries and arteries of the thigh represents a reliable means for the detection of relevant stenoses in patients with peripheral occlusive artery disease. PMID- 11883119 TI - [Cleft formation in a thoracic vertebrae: case report]. PMID- 11883120 TI - [Basilar artery aneurysm]. PMID- 11883121 TI - Fine needle aspiration biopsy of ovarian neoplasm. AB - Twenty cases of ovarian neoplasm (12 non-coelomic and 8 coelomic and 8 epithelial tumors) have been subjected to fine needle aspiration biopsy (FNAB). FNAB yielded adequate material in all those cases and it was possible to accurately classify the tumors in 16 cases (80%). In eight cases of non coelomic epithelial neoplasms preoperative cytological diagnosis helped in conservative surgery. Cytological features of different groups of ovarian tumors are to some extent characteristic. FNAB is a relatively safe and reliable diagnostic procedure. PMID- 11883122 TI - Immunohistochemical localization of transferrin in human breast cancer tissue. AB - The present study was planned to detect the iron binding protein, transferrin (TR) in paraffin sections of the human breast tumors. The distribution of transferrin has been studied in 153 cases (63 benign lesions and 90 malignant tumors). The extent of staining reaction was determined by semiquantitative grading (weak, moderate and consistent). Positivity rate for transferrin was higher (92.2%) in malignant tumors as compared to benign breast lesions (28.5%) with significant p value (P = .0001) for both the groups. The intensity was variable in both the groups, being more intense in the malignant tumors. Tumors with higher grade of malignancy presented consistent positive staining along with the lymph nodes involved. The extent of immunoreactivity revealed a significant positive correlation with axillary lymph node status. However, no significant correlation was found with the age of the patients. Thus the study of transferrin in breast tumors besides being of prognostic significance helps in the further management of malignant lesions of the breast. PMID- 11883123 TI - Prevalence of bacterial vaginosis in antenatal women. AB - Bacterial vaginosis is an established risk factor in pregnant women for premature rupture of membranes and preterm delivery. This study was carried out to find out the prevalence of Bacterial Vaginosis (BV) in antenatal women with vaginal discharge and the effect of treatment with Metronidazole gel on pregnancy outcome. One hundred and fifty symptomatic and fifty asymptomatic women in second trimester of pregnancy in the age group of 20-30 years were included in the study. Gram stained smears of vaginal discharge were examined for evidence of BV with a scoring system by Nugent et al and was found to be positive in 38.5% in symptomatic antenatal women. Intravaginal metronidazole gel application was found to be an effective therapeutic option. Incidence of preterm labour was more in untreated cases. PMID- 11883124 TI - Study of anticardiolipin antibodies in repeated abortions--an institutional experience. AB - A cohort of 178 pregnant women with a history of first or second trimester abortions (2 or more) were the base of present study. In all, other causes of abortion were ruled out except for anti-phospholipid syndrome. Anti-Cardiolipin antibody (ACA) (IgG & IgM) was estimated in the sera samples of all women. Out of 178 women, any one or both immunoglobulins were above the cut off range (> 15.0 units) in 47 (26.4%) while both immunoglobulins were normal in 131 (73.59%) women. Both immunoglobulins were present in only 0.5% women. ACA-IgG alone was present in 11.79% while ACA-IgM alone was present in 14.04% women. We observe from present study that ACA is a major cause of recurrent fetal loss & many pregnancies can be saved if diagnosed & treated adequately. PMID- 11883125 TI - Rapid diagnosis of cholera by coagglutination test. AB - In this study the coagglutination test for the rapid diagnosis of cholera is evaluated in comparison with the conventional culture method. A total of 553 stool specimens were processed from cases of acute gastro-enteritis. The sensitivity and specificity of coagglutination test was 92.77% and 95.65% respectively. The coagglutination test is found to be simple, reliable and rapid method for the diagnosis of cholera. PMID- 11883126 TI - Infective complications of central venous catheters in cardiac surgical patients. AB - Prospective randomised study was conducted over a 24 months period in a cardiac surgical intensive care unit to determine the incidence of infection associated with multilumen venous catheters. The influence of various factors including fever, peripheral blood culture, catheter site, catheter usage for monitoring central venous pressure and/inotrope therapy on infection rates were statistically evaluated. A total of 100 catheters submitted to the Microbiology laboratory were bacteriologically examined. Forty-nine of these were inserted into upper body sites, and 51 were inserted into the femoral vein. Twenty-one were triple-lumen catheters. Catheters were removed when a central line was no longer necessary. Catheter tips were cultured by semiquantitative technique for aerobic and anaerobic bacteria. Bacteremia occurred in 3% of catheter insertions; (Enterococcus faecalis, one; Enterobacter spp. One; Acinetobacter spp., one); and catheter colonisation developed in 24%. Neither catheter colonisation nor catheter related infection were associated with any of the risk factors evaluated. Our data indicates that central venous catheters are safe to use in our patients. The inability to identify "risk factors" for catheter infection emphasise the need to maintain a high index of suspicion. PMID- 11883127 TI - Bacterial indices of drinking water from natural sources. AB - Two hundred forty samples of water from twenty natural sources were subjected to the study for the detection of four bacterial indicators of faecal pollution. The samples were subjected to the detection of coliform, faecal coliform, E. coli, S. faecalis, Cl. welchii and plate count at 37 degrees C and 22 degrees C. All these samples were found unfit for human consumption. MPN for coliforms, E. coli and S. faecalis varied from 3 to 1800 per 100 ml of water, sero to 1800 per 100 ml water and zero to 540 per 100 ml of water respectively. Plate count at 37 degrees C and 22 degrees C varied from 2.5 x 10(3) to > 150 x 10(3) per ml of water. Cl. welchii was detected in 30.4% samples in rainy season and high plate count at 37 degrees C and 22 degrees C was observed during the same period. PMID- 11883128 TI - Recurrent infantile digital fibromatosis--a rare entity. AB - We present here a rare case of Infantile Digital Fibromatosis in a six year old female child who presented with recurrent swelling over the phalanx of left middle finger. PMID- 11883129 TI - Cytodiagnosis of retroperitoneal primitive neuroectodermal tumour (PNET)--a case report. AB - A rare case of Retroperitoneal Primitive Neuro Ectodermal Tumour (PNET) in thirty years old female is presented. Fine needle aspiration cytology (FNAC) characterised by small round cells with scanty cytoplasm, poorly to well formed rosettes with neurofibrillary background; whereas periodic acid-schiff (PAS) rarely react with Primitive Neuro Ectodermal Tumour (PNET), prompted the diagnosis of PNET at cytology itself. Cytological appearances of PNET have been reviewed and the various differential diagnoses are discussed. PMID- 11883130 TI - Sclerosing mediastinitis caused by Aspergillus terreus. AB - A case of Aspergillus terreus causing sclerosing mediastinitis which presented with symptoms of cardiorespiratory compromise and compressive myelopathy is described. The diagnosis was established by culturing and isolating the fungus in pure culture from the tissue and was also confirmed by demonstration of sepcific precipitating antibodies against Aspergillus terreus in patient's serum. PMID- 11883131 TI - Leiomyosarcoma arising from tunica vaginalis testis: a case report. AB - Benign and malignant soft tissue tumors of the paratesticular region i.e. those arising from the testicular tunics, epididymis and spermatic cord are uncommon. Of these, leiomyosarcoma arising from the tunica vaginalis is extremely rare. On extensive computerised search, a single case has been reported till date in the literature. We hereby report one such case because of its rarity. PMID- 11883132 TI - Amyloid goiter--a case report. AB - Microscopic infiltration of the thyroid gland by amyloid is an uncommon but well recognized phenomenon and significant enlargement of the thyroid due to deposition of amyloid is rarely seen. This condition has to be distinguished from other types of goiters and at times from malignancy. In spite of extensive involvement of the gland by amyloid, thyroid function usually remains normal. Here, we describe a rare case of amyloid goiter without amyloid deposits in any other organs of the body. The case presented in an unusual clinical settings of relatively rapid enlargement of thyroid with pressure symptoms and was unassociated with any immuno-proliferative or chronic diseases. PMID- 11883133 TI - Ovarian granulosa cell 'tumorlet' and mature follicles with ectopic decidua in pregnancy--a case report. AB - Non-neoplastic, tumor like lesions are encountered in the ovaries during pregnancy. Of these pregnancy luteoma is the most common lesion mimicking an ovarian neoplasm. Ovarian granulosa cell proliferations are also reported in the ovaries removed during pregnancy as an incidental finding. The granulosa cell proliferation occurs due to the follicular stimulating hormone (FSH) like activity of human chorionic gonadotropin (hCG). We report a case of second gravida who developed uterine atonia, necessitating emergency hysterectomy. Right ovary showed extensive ectopic decidua, numerous well-formed follicles and granulosa cell tumor-like proliferation. Usually follicles are not formed in the ovaries during pregnancy. The same FSH like activity of hCG might have been responsible for the formation and maturation of follicles in this case. PMID- 11883134 TI - Pseudomyxoma peritonei of hernial sac--a case report. AB - A rare presentation of Pseudomyxoma Peritonei of hernial sac is described. The patient was admitted for repair of an inguinal hernia. During herniorraphy large amount of mucinous material was found in hernial sac. Microscopy revealed epithelial glandular cells with bland appearance within mucinous pools. A search for primary remained fruitless. PMID- 11883135 TI - Omental cyst in children presenting as pseudoascites: report of two cases and review of the literature. AB - An omental cyst is a rare intra-abdominal lesion. Absence of characteristic clinical findings make the diagnosis difficult. These cases are reported because of its rarity. The brief review of the features of omental cyst is included in order to increase awareness of this entity. PMID- 11883136 TI - Acute lymphoblastic leukaemia with giant intracytoplasmic inclusion--a case report. AB - Acute Lymphoblastic Leukaemia (ALL) with intracytoplasmic inclusions is a rare and unusual subtype of acute leukaemia. Here we describe a case of ALL with intracytoplasmic inclusions in an adult female. These inclusions stained negative for Myeloperoxidase (MPO), Sudan Black B (SBB) and Alpha-naphthyl acetate estarase (ANAE) and positive for Periodic Acid Schiff (PAS). This case is being presented for its unusual occurrence and to recognise the characteristics of this subtype of ALL to avoid a misdiagnosis of AML (Acute Myeloid Leukaemia). PMID- 11883137 TI - Malignant peritoneal mesothelioma deciduoid or anaplastic variant? Point to ponder. AB - A case of peritoneal mesothelioma displaying unusual morphology, occurring in a 53 year old woman is described. The role of immunohistochemistry and electron microscopy in the evaluation of this tumor is stressed. The appropriate terminology to be used and possible etiologic factor are also discussed. PMID- 11883138 TI - Nerve cell markers in ossifying fibromyxoid tumour of soft parts. AB - Reported herein are two benign ossifying fibromyxoid tumors (OFMTs) of the soft tissues in axilla and terminal phalanx respectively. Both cases on immunohistochemistry (IHC) showed reactivity for vimentin, S-100 protein and glial fibrillary acidic protein (GFAP) antibodies. In addition, a focal/diffuse strong positivity for neurofilament (NF) and neuron specific enolase (NSE) was observed. Electron microscopy in one instance revealed abundant intermediate filaments, primitive cell junctions and a focally present external lamina. In the light of nerve cell differentiation of these tumors and the similarity of IHC profile and EM features of OFMT to a poorly differentiated malignant peripheral nerve sheath tumor (MPNST); it is suggested that OFMT is a variably differentiated PNST rather than a simple Schwannian neoplasm as is believed. PMID- 11883139 TI - Fine needle aspiration cytology, histology and MIB-1 proliferative index in a case of dyshormonogenetic goitre. AB - The fine needle aspiration (FNA) cytologic findings along with histology and MIB 1 proliferative index in a case of dyshormonogenetic goitre is presented. A 12 year old female child from non-endemic zone presented with a isotopically cold solitary thyroid nodule with a history of goitre being present since birth. Past history of any neck irradiation or maternal ingestion of any goitrogen during the antenatal period and family history of goitre were negative. FNA cytology revealed an extremely cellular preparation with predominantly microfollicular pattern without colloid. Nuclei were round to oval, slightly enlarged with evenly distributed chromatin and inconspicuous nucleoli. Larger tissue fragments also showed foci of solid cell groups with nuclear crowding, overlapping and loss of polarity in addition to the prevalent microfollicular pattern. Occasional tissue fragments showed solid groups of trapped follicular epithelial cells in the matrix of fibrocollagenous tissue. Histology showed an intensely hyperplastic follicular cells with nodule formation, irregular fibrosis, pseudo capsular or vascular wall invasion mimicking malignancy. Immunohistochemistry for calcitonin was negative but thyroglobulin was positive. MIB-1 (Ki-67) proliferation index varied from 0.05 to 0.26 (mean 0.13) in the hyperplastic nodules versus 0.9 to 2.1 (mean 1.34) in the hyperplastic solid microfoci scattered amidst the grossly normal appearing thyroid tissue. The possible cytologic diagnostic pitfalls in favor of follicular neoplasm is discussed. PMID- 11883140 TI - Primary malignant giant cell tumour of sacrum--a case report. AB - A rare case of primary malignant GCT of sacrum in a 35-year-old female is reported. PMID- 11883141 TI - Emergence of Vibrio cholerae 0139 in Manipal-coastal Karnataka-South India. PMID- 11883142 TI - Accuracy: key word in quality control in clinical chemistry. PMID- 11883143 TI - Polymerase chain reaction using IS6110 primer to detect Mycobacterium tuberculosis in clinical samples. AB - Nucleic acid amplification using IS6110 primers to detect Mycobacterium tuberculosis in clinical specimens has been extensively used as laboratory tool for the diagnosis for tuberculosis. Despite it's dramatic scientific value in practice, it is not as sensitive as expected for the detection of M. tuberculosis. The results of the study suggest that PCR using 123 bp fragment of DNA belonging to IS6110 is specific (95.6%) but only has a sensitivity of 30% to detect M. tuberculosis in clinical specimens. PMID- 11883144 TI - Emil von Behring and the last hundred years of immunology. PMID- 11883145 TI - Hypothalamo-pituitary-gonadal axis in control of female reproductive cycle. AB - Gonadotropin-releasing hormone (GnRH) secretion from the hypothalamus is pivotal to the regulation of reproductive physiology in vertebrates. The characteristic periodic secretion of gonadotropin releasing hormone (GnRH) from the medial basal hypothalamus (MBH), at the rate of one pulse an hour is essential for the maintenance of the menstrual cycle. These pulses are due to oscillations in the electrical activity of the GnRH pulse generator in the MBH. The GnRH pulse generator is under the influence of an assortment of interactions of multiple neural, hormonal and environmental inputs to the hypothalamus. Hence, a number of conditions such as stress, drug intake, exercise, sleep affect the activity of this pulse generator. Any deviation of normal frequency results in disruption of normal cycle. The cycle can become anovulatory in the hypothalamic lesions and can be restored by exogenous administration of pulsatile GnRH. Of late, studies have shown that pulse generator activity is also maintained by specific metabolic signals meant for energy homeostasis. Studies are in progress to work out cellular basis of GnRH pulse generator's rhythmic activation and role of Ca++ as second messenger for GnRH stimulated gonadotropin release. New concepts are emerging to find the existence of an FSH releasing factor, which independently regulates the activity of FSH. PMID- 11883146 TI - Restoration on tissue antioxidants by fenugreek seeds (Trigonella Foenum Graecum) in alloxan-diabetic rats. AB - The influence of fenugreek seed powder supplementation in the diet on lipid peroxidation and antioxidant status was studied in normal and alloxan-diabetic rats. The protective effect of the aqueous extract of the seeds on the activity of calcium-dependent adenosinetriphosphatase (Ca2+ ATPase) in liver homogenate in the presence of Fe2+/ascorbate in vitro was also investigated. Normal and diabetic rats were provided with a diet supplemented with fenugreek seed powder for 30 days at a dosage of 2 g/kg body weight. The diabetic rats exhibited enhanced lipid peroxidation and increased susceptibility to oxidative stress associated with depletion of antioxidants in liver, kidney and pancreas. However, treatment with fenugreek seed powder normalised the alterations. In normal rats supplementation resulted in increased antioxidant status with reduction in peroxidation. Ca2+ ATPase activity in liver was protected by the aqueous extract to nearly 80% of the initial activity. The findings suggest that the soluble portion of the seeds could be responsible for the antioxidant property. PMID- 11883147 TI - Postnatal changes in the brain lipids, glycolipids and gangliosides of rats exposed to arrack/ethanol during gestation and lactation. AB - Effects of exposure of an alcoholic beverage (arrack and its equivalent quantity of alcohol throughout pregnancy and lactation on brain lipids were investigated. Female rats were exposed to arrack (12.00 ml/kg body weigh/day) and ethanol (4.00 g/kg body weight day) before conception and throughout gestation and lactation. For 21 days pups were nursed by their own mothers, afterwards they were fed normal laboratory feed. We found that the level of cholesterol, phospholipids, triacylglycerols, free fatty acids, cerebrosides, ceramide dihexosides, ceramide polyhexosides, sulfatids,, mono and diglycosyl diglycerides and gangliosides were increased in the brain of 21st and 45th day pups. The alterations in the glycolipid profile of the brain persisted even when pups were not directly exposed to alcohol. These alterations in the glycolipid and ganglioside metabolism may be associated with the developmental abnormalities of the brain seen in FAS. The elevation produced in the glycolipid profile of arrack administered pups were more than that caused by its equivalent quantity of ethanol. This suggested an interaction of congeners in the arrack with the alcohol. PMID- 11883149 TI - In vivo hepatoprotective activity of active fraction from ethanolic extract of Eclipta alba leaves. AB - The alcoholic extract of fresh leaves of the plant Eclipta alba (Ea), previously reported for is hepatoprotective activity was fractionated into three parts to chemically identify the most potent bioactive fraction. The hepatoprotective potential of the fraction prepared from extract was studied in vivo in rats and mice against carbon tetrachloride induced hepatotoxicity. The hepatoprotective activity was determined on the basis of their effects on parameters like hexobarbitone sleep time, zoxazolamine paralysis time, bromosulphaline clearance, serum transaminases and serum bilirubin. Fraction EaII (10-80 mg/kg, p.o.) containing coumestan wedelolactone and desmethylwedelolactone as major components with apigenin, luteolin, 4-hydroxybenzoic acid and protocateuic acid as minor constituents exhibited maximum hepatoprotective activity and is the active fraction for hepatoprotective activity of Eclipta alba leave. The acute toxicity studies have shown that like Ea, Fraction EaII also high safety margin. PMID- 11883148 TI - Protective effects of Indigofera tinctoria L. against D-Galactosamine and carbon tetrachloride challenge on 'in situ' perfused rat liver. AB - The effect of pre-treatment with Indigofera tinctoria (IT) extract against the toxicity of D-Galactosamine (D-GalN) and carbon tetrachloride (CCl4) during 'in situ' perfusion of the liver for 2 hr was studied in rats. Release of LDH and levels of urea in the liver effluent perfusate, was studied and the rate of bile flow was monitored. Perfusion with D-Galactosamine (5 mM) or carbon tetrachloride (0.5 mM) resulted in increased LDH leakage, decreased urea levels in the liver effluent and reduction in bile flow. IT pretreatment (500 mg/kg body weight) in vivo ameliorated D-GalN and CCl4 induced adverse changes towards near normalcy and thereby indicates its hepatoprotective effects in rats. PMID- 11883150 TI - Influence of rice bran oil on serum lipid peroxides and lipids in human subjects. AB - To study the effect of rice bran oil (RBO) on serum lipids and lipid peroxides in human volunteers. Nine healthy volunteers, aged between 42 to 57 years were given 75 ml of RBO thrice daily as the cooking medium with break fast, lunch and dinner for a period of 50 days. At the beginning and at the end of 50 days, 5 ml of blood were drawn from an ante cubital vein. Serum lipids and lipid peroxides levels were estimated from the blood sample. There was a significant decrease in the levels of lipid peroxides, triglycerides, LDL, VLDL, and total cholesterol in human volunteers who switched over to RBO. RBO has evidently antioxidant and antilipidemic activities in human subjects. PMID- 11883151 TI - Role of the medial preoptic area in thermal preference of rats. AB - This study was conducted to find out whether the medial preoptic area (mPOA) plays a role in the selection of ambient temperature by rats. Adult male Wistar rats were kept in an environmental chamber having three interconnected compartments, maintained at three different temperatures (18 degrees, 24 degrees and 30 degrees C) in which the animals could move freely from one compartment to the other. Normal rats preferred to stay at the chamber maintained at 24 degrees C for most of the time, during day and night. The temperature preference shifted to 30 degrees C after the mPOA of these rats had been lesioned by local administration of 5 micrograms of N-methyl D-aspartic acid (NMDA) in 0.2 microliter distilled water. The results of the study suggest that the mPOA acts as a fine tuning center for homeostatic regulation of thermal balance, including selection of appropriate thermal environment. It is proposed that after the mPOA lesion, the animal cannot assess properly the energy status of the body and thereby prefers a higher ambient temperature. PMID- 11883152 TI - Effect of fasting on the intestinal absorption of D-glucose and D-xylose in rats in vivo. AB - The present study was planned to elucidate the role of fasting on the intestinal absorption of monosaccharides particularly--glucose and xylose in inbred female albino rats. Rats (weighing 250-300 grams) were divided into three groups. One group of rats served as control while the other two were experimental. One of the experimental groups was starved for 48 hours while the other for a period of 72 hours. It was found that fasting for 72 hours causes an overall increase in absorption of glucose from small intestine. Forty-eight hours of fasting caused a significant increase in glucose absorption from distal ileum only. Increase in the glucose absorption in fasting from small intestine can well be explained on the basis of a reduction in glucose metabolism in general as an adaptation to starvation so as to leave more glucose for cerebral metabolism. No significant changes, whatsoever, were encountered with xylose absorption in fasting animals. PMID- 11883153 TI - Physical fitness: a longitudinal study among Muslim children of Bijapur (Karnataka). AB - Aerobic capacity or maximum oxygen uptake capacity (VO2 max) has been widely considered to be reliable and valid measure of cardio respiratory fitness. Persons possessing higher values and have the capacity to yield larger amounts of energy, are capable of performing better in athletic and other field activities. Seventy school going children from the Muslim community of Bijapur (Karnataka) aged 12-16 years (means +/- SEM = 14.33 +/- 0.94), volunteered for this study. Their height (cm) and weight (kg) were measured as physical anthropometry and Body mass index (BMI) was calculated (kg/m2). VO2max (ml.kg-1.min-1) was determined by applying the step test study of Margaria et al. The Physical fitness index (PFI) of the subjects were assessed by Harvard Step Test. The physiological endurance measured as VO2max (ml.kg-1.min-1) was found to be 34.31 +/- 2.44 S.E.M, which is lower in comparison to their Caucasian counterparts but nearly similar when compared with their Indian counterparts. The present study reveals that VO2max significantly correlates with BMI and PFI score. The present study also reveals that 27.2%, 20.07%, 15.77%, 14.37% and 22.87% of the subjects are in excellent, very good, good, average and poor classifications of fitness level respectively. PMID- 11883154 TI - Ventricular nociception induced vesicular motility and urine flow: their relationship. AB - Heart acts as an important reflexogenic organ. Reflex urination and defaecation are two of the most important visceral symptoms observed in patients with myocardial ischaemia, infarction etc. In experimental animals also ventricular nociceptor stimulation by left anterior descending coronary artery (LAD) occlusion and nicotine application causes biphasic changes in urinary bladder movement and urine flow. Aim of the present study is to elucidate if there is any correlation between urine formation by the kidneys and movement of the urinary ladder under such experimental conditions. The experiments performed on intact cats show apparent coincidence of the two events. But, subsequent experiments following denervation of vagi and inferior cardiac nerve (ICN), spinal transaction and decerebration experiments indicate that these two are separate events. Further, experiments with different neurotransmitter blockers indicate that ventricular nocieptor induced urine formation and urinary bladder movements are two separate reflex responses and not dependent on each other. PMID- 11883155 TI - Autonomic functions in Buerger's disease. AB - One of the pathophysiological features in Buerger's disease, i.e. thromboangitis obliterans (TAO), is vasospastic phenomena. So autonomic reactivity was evaluated in 12 patients of Buerger's disease (1-6 years duration) and compared to age and sex matched controls (nonsmokers). Basal heart rate was significantly (P < 0.001) higher without any variation in blood pressure in TAO group compared to controls. Valsalva ratio (P < 0.01) and 30:15 ratio (P < 0.001) were increased without any effect on E:I ratio in TAO group versus controls. On head up tilt (HUT), there was significant (P < 0.001) fall in blood pressure in TAO group compared to controls. On cold pressure test (CPT), systolic blood pressure was reduced significantly (P < 0.01) in TAO group than that of controls, however, diastolic blood pressure showed no change in two groups. Responses indicate towards lower sympathetic reactivity in Buerger's patients. PMID- 11883156 TI - Aspirin modulates the anticonvulsant effect of diazepam and sodium valproate in pentylenetetrazole and maximal electroshock induced seizures in mice. AB - Release of prostaglandins in brain after spontaneous and experimentally induced seizures, has been demonstrated. The possible role of prostaglandins in modulation of seizure activity is still inconclusive. In the present study, the effects of aspirin and its interaction with the anticonvulsants (diazepam and sodium valproate) were studied in pentylenetetrazole (PTZ) and maximal electroshock (MES) induced seizures in mice. Aspirin 50, 100, and 500 mg/kg, i.p. was administered 45 min before the pentylenetetrazole (60 mg/kg, i.p.) and MES (60 mA, 0.2 s duration via car clip electrodes) challenge. In MES seizures significant protection was seen with aspirin 100 mg/kg where as higher dose of aspirin 500 mg/kg was required to elicit maximum protection against PTZ seizures. Sub anticonvulsant dose of sodium valproate 150 mg/kg, i.p. and aspirin 50 mg/kg i.p. showed complete protection in MES seizures and the same dose of sodium valproate offered superior protection in PTZ seizures than either drug used alone. When mice were pretreated with combination of diazepam 0.5 mg/kg and aspirin 50 mg/kg protection was significantly enhanced in PTZ seizures. However, aspirin did not show any significant protection with subanticonvulsant dose of diazepam against MES seizures. The present study suggests that prostaglandins may have anticonvulsant potential and also may have modulatory effect on anticonvulsant effect of conventional antiepileptic drugs. PMID- 11883157 TI - Hepatoprotection by Elephantopus scaber Linn. in CCl4-induced liver injury. AB - The efficacy of the medicinal plant Elephantopus scaber Linn. (Asteraceae), to prevent carbon teterachloride (CCI4)-induced chronic liver dysfunction in the rats was examined by determining different biochemical markers in serum and tissues. In serum, liver function marker enzymes like aspartate aminotrasferase (AST), alanine aminotrasferase (ALT), alkaline phosphatase (ALP) and also protein were evaluated. The concentrations of total lipid, cholesterol and phospholipids were studied in serum and the different tissues. The concentration of serum triglycerides was also studied. The biochemical changes induced by CCI4 in different tissues particularly in the liver tissue improved following treatment with E. scaber Linn. The results suggest the hepatoprotective effect of this medicinal plant. PMID- 11883158 TI - Effect of chronic vibration on the immune state of albino rats. AB - A study was undertaken to investigate the effects of whole body chronic vibration on the immune system. Albino rats were exposed to whole body horizontal vibration acceleration 5.0 g, frequency 20 Hz for 3 hours per day for 3 months and changes were observed in plasma corticosterone level, total leucocyte count and differential leucocyte count. Neutrophil functions were accessed by candida phagocytosis and Nitroblue tetrazolium reduction test. The total leucocyte count was significantly decreased. A marked lymphopenia was observed in the differential count of the leucocytes. A significant increase in the plasma corticosterone level, candida phagocytosis and Nitroblue tetrazolium reduction was observed, indicating chronic whole body vibration to be a potent stressor in albino rats. PMID- 11883159 TI - Effect of yogic practice on pulmonary functions in young females. AB - During recent years, a lot of research work has been done to show the beneficial effects of yoga training. The present study was undertaken to assess the effects of yogic practice on some pulmonary functions. Sixty healthy young female subjects (age group 17-28 yrs.) were selected. They had to do the yogic practices daily for about one hour. The observations were recorded by MEDSPIROR, in the form of FVC, FEV-1 and PEFR on day-1, after 6 weeks and 12 weeks of their yogic practice. There was significant increase in FVC, FEV-1 and PEFR at the end of 12 weeks. PMID- 11883160 TI - EMG characteristics and fibre composition: study on rectus femoris of sprinters and long distance runners. AB - The study was conducted on 9 sprinters and 5 long distance runners to investigate the difference in power spectral characteristics of rectus femoris muscle and the feasibility of using electromyographic techniques in categorization of muscle groups in slow dominant and fast dominant types. EMG signal was recorded, after digitization at 4 KHz, from rectus femoris muscle during isometric knee extension (at maximum voluntary contraction level) until fatigue. Digitized signal was processed for Fast Fourier Transform and Root Mean Square (RMS) voltage. Significant difference (P < 0.05) was found in RMS voltage between sprinters and long distance runners. Both groups showed decline in Mean Power Frequency (MPE) and rate of decline in sprinters was rapid. Normalized MPF showed better discrimination between the two groups. It is concluded that the EMG response observed in this study was possibly a result of differences in the muscle fibre composition of the athletes. EMG study using spectral characteristics would be useful in categorizing the sports persons in terms of suitability of the events. PMID- 11883161 TI - Antiepileptic activity of Panax ginseng against pentylenetetrazole induced kindling in rats. AB - In the present study, Panax ginseng was evaluated for its antiepileptic activity against pentylenetetrazole (PTZ) induced chemical kindling in rats. PTZ was injected at the dose of 30 mg/kg, i.p. on alternate days and the occurrence of generalized tonic clonic convulsions were considered as the end point. One group received Panax ginseng every day, at a dose of 100 mg/kg, 30 min prior to PTZ injection whereas the other group received an equal volume of distilled water to serve as control. In a separate group the rats were evaluated for motor performance tests after Panax ginseng. The rats treated with Panax ginseng showed significant protection as compared to vehicle treated PTZ injected rats. The study suggests to potential of Panax ginseng against seizures. PMID- 11883162 TI - Effect of common anti-epileptic drugs on cognition in schoolchildren with epilepsy. AB - This study was conducted to observe the effect of some commonly used anti epileptic drugs (AEDs), on cognition, in 118 school going children with epilepsy, in an age range of 9-12 yrs., (Mean 10.4 +/- 1.7 yrs.). For comparison, 28 healthy, age and sex matched schoolchildren served as controls. After a clinical, electrophysiological and radiological evaluation, the cognitive functions were assessed in both groups, using a modified Wechsler's Intelligence Scale. It was observed that cognition was impaired in only 2.5% of children with epilepsy, there being no relationship between cognitive performance and the type of AED used. It is concluded that cognitive functions are impaired in only a limited number of children with epilepsy and effect of phenobarbitone and phenytoin on cognitive functions is comparable to carbamazepine and sodium valproate, particularly when demand of task is not very high. PMID- 11883163 TI - Extra systoles in the frog heart. PMID- 11883164 TI - Global measles mortality reduction and regional elimination, 2000-2001. Part II. PMID- 11883165 TI - Recommended composition of influenza virus vaccines for use in the 2002-2003 season. PMID- 11883166 TI - Case management models, not staffing formulas, dictate caseload. PMID- 11883167 TI - Knowledge of home care key for case managers. PMID- 11883169 TI - Bioterror response requires targeted disaster plan. PMID- 11883168 TI - How to appraise your teamwork performance. PMID- 11883170 TI - Find optimal staffing for outpatient surgery. PMID- 11883171 TI - Ambulatory programs call for different type of CQI. PMID- 11883172 TI - [The role of behavioral sciences in the Hungarian and international medical education: overview and possibilities]. AB - The unilateral bio-medical approach cannot be effective in the prevention and effective treatment of chronic disorders of great epidemiological significance, because the behavioural risk factors are strongly influenced by psychosocial factors too. After the change of the political system in Hungary the most important step in curriculum development was establishing the institutes of behavioural sciences. However, the share of behavioural sciences nowhere exceeds 3% of the curriculum. Yet it has an important role in bridging the gap between the natural and social sciences. Built on the firm basis of natural sciences, the behavioural sciences complement this foundation with aspects of social sciences which emphasize the psychological needs of the patients as well as the psychosocial determinants of health and diseases. The most significant field of behavioural sciences is medical psychology but according to the latest reports on medical education development communication, medical sociology, medical anthropology and bioethics have become increasingly important disciplines in developing medical competence. These fields are organized into an integrated process, arching over the six years of medical training. The present paper gives an overview of the situation and perspectives of teaching behavioural sciences at medical universities. PMID- 11883173 TI - [Immune thrombocytopenic purpura (ITP)]. AB - Immune thrombocytopenic purpura (ITP) belongs to the major classical entities of clinical haematology. Diagnosis is still based on documentation of "megakaryocytic thrombocytopenia" and exclusion of other factors, diseases. The childhood (mainly acute) and adult type (mainly chronic) seem to differ substantially, and run different prognosis as well as response to therapeutic measures. A lasting remission or cure is uncommon with corticosteroids alone in the chronic cases, so splenectomy is frequently necessary and indicated. Standard therapy is poorly defined or established in splenectomy refractory cases, which situation requires experience, therapeutic skills and special care. There are many promising efforts to spare splenectomy, especially in younger patients. New standards are also recommended in ITP crisis situations (wet purpura, pregnancy and delivery, splenectomy, etc.). Standard as well as innovative approaches, focusing on clinical care, are briefly evaluated and reviewed in this report. PMID- 11883174 TI - [Angioneurotic edema induced by angiotensin converting enzyme inhibitors]. AB - Angioneurotic oedema is one of rare side effects of angiotensin converting enzyme inhibitors, its incidence is around 0.1-0.2%. Angio-oedema most commonly develops in the first 4 weeks of the treatment, but it can be observed later, after several months or even years. The association between the oedema and the drug intake can be difficult to recognize if the oedema is of delayed type and because the attacks can disappear spontaneously without discontinuation of the drug. The angioneurotic oedema is tend to be worsening during the treatment, and finally the obstruction of the upper respiratory tract can be fatal. The affected sites are the face, lips, tongue, upper respiratory tract, and the oedema can also develop in the gastrointestinal tract with abdominal pain and diarrhea, which can be misdiagnosed. The pathomechanism is thought to be rather biochemical than immunological. The pathogenetic factors are under investigation nowadays, but the increased level of bradykinin seems to be the most important factor. Authors treated 248 patients with angioneurotic oedema in the Department of Dermatology (Semmelweis Hospital, Miskolc) between January of 1997 and December of 2000, 44 patients took angiotensin converting enzyme inhibitors, and 16 patients were suspected as suffering from angio-oedema induced by this drug. All of the patients remained symptom-free after the adequate treatment and discontinuation of the suspected drug. Authors describe the clinical picture of the angio-oedema, the risk factors, and the contraindications of the angiotensin converting enzyme inhibitor treatment. PMID- 11883175 TI - [The role of perisinusoidal (Ito) cells in the course of hepatocellular carcinoma: light and electron microscopic data to the pathogenesis of hepatocellular carcinoma]. AB - The clinical history of a 49 year old female patient suggested a multifocal, rapidly progressive liver disease of one month duration, apparently due to metastatic tumour. An open needle biopsy of the liver revealed a primary hepatocellular carcinoma of low grade malignancy; the diagnosis was confirmed by histological, immunohistochemical and electron microscopic studies. Besides the ultrastructural examination of the liver biopsy disclosed an unusually marked proliferation of perisinusoidal (Ito)-cells. The authors assume that the myofibroblast proliferation and transformation of Ito-cells in the noncirrhotic liver led to the formation of multifocal areas. The perisinusoidal cell proliferation was presumably due to vitamin A intoxication caused by an extreme vegetarian diet (daily consumption of large amounts of carrot juice for years, as disclosed by a retrospectively obtained history). It is assumed that the vitamin A abuse, and perisinusoidal cell proliferation may have promoted the unusually rapid progression of the multifocal, but histologically low grade hepatocellular tumour. Spectacular clinical improvement could be observed after chemotherapy, combined with local hyperthermic treatment. Presumably, the change in diet (cessation of excessive retinol and carotene intake) also may have had a beneficial effect. After one year the clinical course suggests a slower progression of tumour growth which would be more in keeping with the prognosis based on the histologic appearance of the low grade hepatocellular carcinoma. This patient's case illustrates the importance of electron microscopy supplementing diagnostic histological and immunohistochemical examinations. PMID- 11883176 TI - [Surgical treatment of giant basilar artery aneurysm with induced hypothermia and circulatory arrest]. AB - Surgical management of giant and complex posterior circulation aneurysms continues to be a technically difficult task with high operative morbidity. To minimize morbidity we have used cardiopulmonary bypass and circulatory arrest for the treatment of a giant basilar aneurysm. A 48-year-old woman presented with sudden headache. Magnetic resonance angiography revealed a giant basilar aneurysm. On the 2nd hospital day she developed right sided hemiparesis and cranial nerve deficits as a result of the second rupture of the aneurysm. The aneurysm was successfully treated and no significant neurological complications were related to this technique. This initial experience indicates that patients with giant posterior circulation aneurysm that cannot be treated using conventional techniques might benefit from a surgical approach that included the use of deep hypothermic circulatory arrest. PMID- 11883177 TI - [Data on the medical historian Laszlo Nemeth]. PMID- 11883178 TI - Catholic system, physicians collaborate on pneumonia care. PMID- 11883179 TI - Demo produces stunning results in care of complex patients. PMID- 11883180 TI - Blood cultures, antibiotic therapy boost outcomes in septicemia. AB - Septicemia is a common diagnosis among nursing home patients, which can result in hospital admission and often unnecessarily long lengths of stay, incorrect coding, and repeat readmissions. At St. Joseph's Hospital in Tampa, FL, a process improvement team (PIT) decided to focus its efforts on the management of septicemia patients with results you may want to emulate. PMID- 11883181 TI - New guidelines help providers refine Parkinson's strategy. PMID- 11883182 TI - Reovirus receptors and apoptosis. PMID- 11883183 TI - The avian reovirus genome segment S1 is a functionally tricistronic gene that expresses one structural and two nonstructural proteins in infected cells. AB - The avian reovirus S1 gene contains three partially overlapping, out-of-phase open reading frames (ORFs) that the highly conserved in all avian reovirus strains examined to date. The three S1 ORFs of the avian reovirus strain S1133 were individually expressed in bacterial cells, and their purified translation products used as antigens to raise specific polyclonal antibodies. With these antibodies we were able to demonstrate that all three S1 ORFs from different avian reovirus strains are translatable in infected cells. Proteins p10 and p17, which are specified by ORF1 and ORF2, respectively, are nonstructural proteins which associate with cell membranes, whereas ORF3 directs the synthesis of protein sigma C, a structural oligomeric protein responsible for cell attachment. While intracellular synthesis of protein sigma C was demonstrated a long time ago and that of protein p10 was reported recently, this is the first time that expression of the S1 ORF2 has been demonstrated experimentally. Thus, the previously reported coding capacity of the avian reovirus genome is now expanded to 14 proteins, of which ten are structural (lambda A, lambda B, lambda C, microA, microB, microBC, microBN, sigma A, sigma B, and sigma C) and four are nonstructural (microNS, sigma NS, p17, and p10). Finally, protein p10, but not p17 or sigma C, induces cell-cell fusion when transiently expressed in mammalian cells, supporting a previously published observation that the polypeptide encoded by the S1 ORF1 plays an important role in the syncytial phenotype displayed by avian reoviruses. PMID- 11883184 TI - Critical role for SV40 small-t antigen in human cell transformation. AB - Defining the ability of simian virus 40 (SV40) to transform human cells has become of even greater importance with the increased understanding that this virus may play a role in some human malignancies. This report documents the requirement for viral small-t (ST) antigen in large-T (LT)-driven transformation of primary fibroblasts, a requirement that cannot be met by a well-known oncogene, c-Ha-ras (EJ-ras), which can cooperate with LT in rodent systems. The cellular gene telomerase is not essential for transformation, although transformed clones are not immortal without it. Similarly, an immortal mesothelial cell line has been developed using LT and telomerase. Immortalized mesothelial cells are morphologically normal, but can be transformed by introduction of ST, or ST + ras, but not by ras alone. It is likely that ST will be required along with LT for transformation of most human cell types. PMID- 11883185 TI - Characterization of the developmental switch region of bacteriophage P2 Hy dis. AB - In this work, the DNA sequence of the transcriptional switch that affects the development of the P2 Hy dis bacteriophage was determined. The switch contains two face-to-face-located promoters and two repressors, Cox and C. The locations of the Pc and Pe promoters were determined by primer extension analysis. The P2 Hy dis homolog of the P2 multifunctional Cox protein was shown to be able to substitute for P2 Cox in repression of the P2 Pc promoter, excision of the P2 prophage, and activation of the satellite phage P4 PLL promoter. A directly repeated sequence, flanking the--35 region of the Pe promoter, was found to be important for C repressor binding as well as for repression. The P4 E protein was shown to derepress the developmental switch of P2 Hy dis in a plasmid-based derepression assay. PMID- 11883186 TI - The baculovirus transcriptional transactivator ie0 produces multiple products by internal initiation of translation. AB - Ie0 is the only gene of the baculovirus Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) that is known to be spliced. In this study, cDNAs of ie0 were isolated, cloned, and sequenced. It was observed that IE0 contains 35 amino acids (aa) added to the N-terminus of IE1. In addition, it was found that the leader sequence of ie0 contains a 4-aa minicistron. To functionally characterize IE0, ie0 cDNAs were expressed under control of either the ie1 or the ie0 promoter. Unexpectedly, examination of ie0 translation products revealed that the predominant product from ie0 mRNAs was not IE0, but IE1. Mutation analysis showed that IE1 translation was preferentially initiated from either of two AUGs found in the first 15 nucleotides (nt) of the ie1 ORF that are internal to the ie0 ORF. It is unknown whether the internal translation initiation occurs via a leaky scanning mechanism or by an internal ribosomal entry site. Transactivation analysis with constructs that had point mutations in the ie1 AUGs and were translated only as IE0 revealed that OpMNPV IE0 is a 14- to 15-fold stronger transactivator than IE1. IE0 was also shown to be autoregulatory and to transactivate early genes in an enhancer-independent or -dependent manner. These results suggest that differential expression of baculovirus early genes can be obtained by coexpression of IE0 and IE1 in infected cells, which may permit subtle regulation of specific sets of viral genes. PMID- 11883187 TI - The hepatitis C virus core protein interacts with NS5A and activates its caspase mediated proteolytic cleavage. AB - Viral proteins interact with one another during viral replication, assembly, and maturation. Systematic interaction assays of the hepatitis C virus (HCV) proteins using the yeast two-hybrid method have uncovered a novel interaction between core and NS5A. This interaction was confirmed by in vitro binding assays, and coimmunoprecipitation in mammalian cells. Core and NS5A are also colocalized in COS-7 cells. Interestingly, NS5A is cleaved to give specific-size fragments, when core is coexpressed in mammalian cells. Overexpression of core produced many dying and rounded cells and effects such as DNA laddering and the truncation of poly(ADP-ribose) polymerase 1 (PARP1), both indicators of apoptosis. These observations led us to investigate the link between the induction of apoptosis by core and the cleavage of NS5A. The proteolysis of NS5A and these apoptotic events can be inhibited by caspase inhibitor, Z-VAD, indicating that core induces apoptosis and the cleavage of NS5A by caspases. In cells infected by the HCV, core may provide the intrinsic apoptotic signal, which produces truncated forms of NS5A. The biological function of core-NS5A interaction and the downstream effect of NS5A cleavage are discussed. PMID- 11883188 TI - Expression of human and macaque type I IFN transgenes interferes with HSV-1 replication at the transcriptional and translational levels: IFN-beta is more potent than IFN-alpha 2. AB - A study was undertaken to compare the efficacy of plasmid constructs encoding human IFN-alpha 2 and IFN-beta and macaque IFN-beta against herpes simplex virus type 1 in transfected cells. All type I IFN transgenes significantly reduced viral titers in transfected cells by 3 logs. Human IFN-alpha 2-transfected cells produced significantly more IFN (2274 pg/ml) in comparison to IFN-beta transfected cells (134-165 pg/ml). Viral lytic gene transcript and viral protein levels were lower in IFN-beta- versus IFN-alpha 2-transfected cells, which coincided with elevated PKR and OAS transcript levels and increased total STAT1 and phosphorylated STAT1 (Y701) protein levels in the IFN-beta-transfected cells. Although comparable viral titers were recovered in IFN-alpha 2 and IFN-beta plasmid-transfected cells, IFN-alpha 2 plasmid-transfected cells exhibited significantly more cytopathic effect compared to the IFN-beta transgene transfected cells. In addition, IFN-alpha 2 transgene-transfected, infected cells displayed a cell cycle profile similar to that of vector-transfected, infected cells, whereas IFN-beta plasmid-transfected cells displayed a profile similar to uninfected control. Collectively, the results indicate that human IFN-beta is superior to IFN-alpha 2 in antagonizing herpes simplex virus type 1 infection. PMID- 11883189 TI - Movement proteins (BC1 and BV1) of Abutilon mosaic geminivirus are cotransported in and between cells of sink but not of source leaves as detected by green fluorescent protein tagging. AB - Two movement proteins (BV1 and BC1) facilitate the intra- and intercellular transport of begomoviruses in plants. In contrast to other geminiviruses the movement protein BC1 of Abutilon mosaic virus (AbMV) remained in the supernatant after centrifuging plant extracts at 20,000 g. To test whether this unusual behavior results from a distinct intracellular distribution of the protein, the BC1 gene has been fused to the gene of green fluorescent protein (GFP). The resulting plasmids were delivered into nonhost plants (Allium cepa) as well as into mature and immature cells of host plants (Nicotiana tabacum, N. benthamiana) by biolistic bombardment for transient expression in planta. BC1 directed GFP to two different cellular sites. In the majority of nonhost cells as well as in mature cells of host leaves, BC1 was mainly localized in small punctate flecks at the cell periphery or, to a lesser extent, around the nucleus. In sink leaves of host plants, GFP:BC1 additionally developed disc-like structures in the cell periphery. Cobombardment of GFP:BC1 with its cognate infectious DNA A and B did not change their subcellular distribution patterns in source leaves but led to the formation of peculiar needle-like structures in sink leaves. The nuclear shuttle protein (BV1) of AbMV accumulated mainly inside the nuclei as shown by immunohistochemical staining and GFP tagging. In sink cells of host plants it was mobilized to the plasma membrane and to the nucleus of the neighboring cell by coexpressed BC1, GFP:BC1, BC1:GFP, or after cobombardment with the cognate viral DNA. Only under these conditions were GFP:BC1 and BC1:GFP also found in the recipient cell. PMID- 11883190 TI - Leader protein of encephalomyocarditis virus binds zinc, is phosphorylated during viral infection, and affects the efficiency of genome translation. AB - Encephalomyocarditis virus (EMCV) is the prototype member of the cardiovirus genus of picornaviruses. For cardioviruses and the related aphthoviruses, the first protein segment translated from the plus-strand RNA genome is the Leader protein. The aphthovirus Leader (173-201 amino acids) is an autocatalytic papain like protease that cleaves translation factor eIF-4G to shut off cap-dependent host protein synthesis during infection. The less characterized cardioviral Leader is a shorter protein (67-76 amino acids) and does not contain recognizable proteolytic motifs. Instead, these Leaders have sequences consistent with N terminal zinc-binding motifs, centrally located tyrosine kinase phosphorylation sites, and C-terminal, acid-rich domains. Deletion mutations, removing the zinc motif, the acid domain, or both domains, were engineered into EMCV cDNAs. In all cases, the mutations gave rise to viable viruses, but the plaque phenotypes in HeLa cells were significantly smaller than for wild-type virus. RNA transcripts containing the Leader deletions had reduced capacity to direct protein synthesis in cell-free extracts and the products with deletions in the acid-rich domains were less effective substrates at the L/P1 site, for viral proteinase 3Cpro. Recombinant EMCV Leader (rL) was expressed in bacteria and purified to homogeneity. This protein bound zinc stoichiometrically, whereas protein with a deletion in the zinc motif was inactive. Polyclonal mouse sera, raised against rL, immunoprecipitated Leader-containing precursors from infected HeLa cell extracts, but did not detect significant pools of the mature Leader. However, additional reactions with antiphosphotyrosine antibodies show that the mature Leader, but not its precursors, is phosphorylated during viral infection. The data suggest the natural Leader may play a role in regulation of viral genome translation, perhaps through a triggering phosphorylation event. PMID- 11883191 TI - Isolation and characterization of two viruses with large genome size infecting Chrysochromulina ericina (Prymnesiophyceae) and Pyramimonas orientalis (Prasinophyceae). AB - Two lytic viruses specific for Chrysochromulina ericina (Prymnesiophyceae) and for Pyramimonas orientalis (Prasinophyceae) were isolated from Norwegian coastal waters in June 1998. The lytic cycle was 14-19 h for both viruses; the burst size was estimated at 1800-4100 viruses per host cell for the Chrysochromulina virus and 800-1000 for the Pyramimonas virus. Thin sections of infected cells show that both viruses replicate in the cytoplasm and that they have a hexagonal cross section, indicating icosahedral symmetry. The Chrysochromulina virus had a particle size of 160 nm and a genome size of 510 kbp; the size of the major polypeptide was 73 kDa. The Pyramimonas virus had a particle size of 220 x 180 nm and a genome size of 560 kbp; the size of the major polypeptide was 44 kDa. The genome sizes of these viruses are among the largest ever reported for viruses and they are larger than the minimum required for cellular life. The Chrysochromulina virus clone CeV-01B and the Pyramimonas virus clone PoV-01B described in this study have several properties in common with other viruses infecting microalgae, suggesting that they belong to the Phycodnaviridae. PMID- 11883192 TI - Prolonged E55+ retrovirus expression in aged mice is associated with a decline in the anti-virus immune response. AB - E55+ murine leukemia retrovirus (E55+ MuLV) infection of young and aged C57BL/6 (B6) mice was used to investigate the relationship between increased incidences of infection and decreased immune responsiveness of elderly individuals. Young mice decreased E55+ MuLV burden to below detectable levels by 8 weeks postinfection (p.i.). In contrast, virus burden in aged mice did not reach undetectable levels until 20 weeks p.i. A significant T cell proliferative response to E55+ MuLV was detected from 2 to 12 weeks p.i. in young mice, but was never observed in aged mice. Both age groups demonstrated significant E55+ MuLV specific T-cell-mediated cytotoxic responses at 3 and 4 weeks p.i. and virus neutralizing antibody titers at 2, 4, 8, and 12 weeks p.i. In both cases, responses were consistently higher in young mice (P < 0.04 and P < 0.02, respectively). These results demonstrate that the observed delay in E55+ MuLV clearance by aged mice is associated with an age-related decrease in the immune response to the virus. PMID- 11883193 TI - Cloning, expression, and crystallization of the fusion protein of Newcastle disease virus. AB - We have recently reported the X-ray crystal structure of a fragment of the fusion protein (F) of Newcastle disease virus (NDV). This work describes the methodology involved in the production and crystallization of that protein in recombinant form. The full-length cDNA of NDV-F was cloned and the ectodomain expressed in both CHO-K1 and Lec-3.2.8.1 cells. The recombinant protein, secreted as a single chain polypeptide F0', was purified using a c-myc antibody affinity column followed by gel filtration chromatography. Electron microscopic imaging showed the F0' product to consist of unaggregated club-shaped particles. Trypsin treatment of F0' could be used to produce disulfide-linked F2 and F1' chains. However, imaging revealed extensive rosette-like aggregation of the trypsin treated material, indicative of a conformational change. Only the non-trypsin treated product was thus suitable for crystallization and two crystal forms were obtained, diffracting to ca. 3.5 and 4.0 A, respectively. Both crystal forms were used in the structure determination. PMID- 11883194 TI - Glutamic residue 438 within the protease-sensitive subdomain of HIV-1 reverse transcriptase is critical for heterodimer processing in viral particles. AB - The biological form of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is a heterodimer consisting of two polypeptides, p66 and p51, which have identical N-termini. The p51 polypeptide is generated by action of viral protease cleaving the p66 polypeptide between residues Phe440 and Tyr441. Dimerization has been mostly studied using bacterially purified RT bearing amino acid changes in either subunit, but not in the context of HIV-1 particles. We introduced changes of conserved amino acid residues 430-438 into the protease sensitive subdomain of the p66 subunit and analyzed the reverse transcriptase processing and function using purified variants and their corresponding HIV-1 recombinant clones. Our mutational analysis shows that the conserved Glu438 residue is critical for proper heterodimerization and function of virion associated RT, but not of bacterially expressed RT. In contrast, the conserved Glu430, Glu432, and Pro433 residues are not important for dimerization of virion associated RT. The network of interactions made by the Glu438 carboxyl group with neighboring residues is critical to protect the Phe440-Tyr441 from cleavage in the context of the p66/p51 heterodimer and may explain why the p66/p51 is not processed further to p51/p51. PMID- 11883195 TI - Phenotypic and molecular analyses of yellow fever 17DD vaccine viruses associated with serious adverse events in Brazil. AB - The yellow fever (YF) 17D virus is one of the most successful vaccines developed to data. Its use has been estimated to be over 400 million doses with an excellent record of safety. In the past 3 years, yellow fever vaccination was intensified in Brazil in response to higher risk of urban outbreaks of the disease. Two fatal adverse events temporally associated with YF vaccination were reported. Both cases had features similar to yellow fever disease, including hepatitis and multiorgan failure. Two different lots of YF 17DD virus vaccine were administered to the affected patients and also to hundreds of thousands of other individuals without any other reported serious adverse events. The lots were prepared from the secondary seed, which has been in continuous use since 1984. Nucleotide sequencing revealed minor variations at some nucleotide positions between the secondary seed lot virus and the virus isolates from patients; these differences were not consistent across the isolates, represented differences in the relative amount of each nucleotide in a heterogeneous position, and did not result in amino acid substitutions. Inoculation of rhesus monkeys with the viruses isolated from the two patients by the intracerebral (ic) or intrahepatic (ih) route caused minimal viremia and no clinical signs of infection or alterations in laboratory markers. Central nervous system histological scores of rhesus monkeys inoculated ic were within the expected range, and there were no histopathological lesions in animals inoculated ih. Altogether, these results demonstrated the genetic stability and attenuated phenotype of the viruses that caused fatal illness in the two patients. Therefore, the fatal adverse events experienced by the vaccinees are related to individual, genetically determined host factors that regulate cellular susceptibility to yellow fever virus. Such increased susceptibility, resulting in clinically overt disease expression, appears to be extremely rare. PMID- 11883196 TI - Herpes simplex virus gene products required for viral inhibition of expression of G1-phase functions. AB - HSV infection blocks G1 events in the cell cycle and arrests host cell growth in the G1 phase. To further define the mechanism of the effect and determine the viral gene product(s) responsible, we examined various mutant viruses for their effects on cell cycle regulatory proteins (pRb, cyclin D1, and cdk4) and on cell cycle progression into S phase. Unlike the wild-type virus, the ICP27 mutant virus was defective for blocking the phosphorylation of pRb proteins, and the normal pRb pattern was restored in cells infected with a rescued virus. The virion host shutoff (vhs) function, DNA replication, and late gene functions were not required for the virus-induced effects on pRb protein. BrdU incorporation in synchronized HSV-infected cells showed that ICP27 was required for blocking the cell cycle in the G1 phase. Furthermore, ICP27, ICP4, ICP0, and vhs were required for blocking the induction of the G1 cell cycle regulators cyclin D1 and cdk4 in HSV-infected cells. Both ICP27 and the vhs function contributed to the reduction of cyclin D1 mRNA levels in HSV-infected cells: These results provide evidence that HSV-1 ICP27 protein is essential for viral inhibition of G1-phase functions and that certain other HSV proteins are required for some of the viral effects on the cell cycle. Finally, these results show that HSV-1 ICP27 and vhs act jointly to reduce host mRNA levels in infected cells. PMID- 11883197 TI - Role of ATP in influenza virus budding. AB - Influenza viruses bud from the plasma membrane of virus-infected cells. Although budding is a critical step in virus replication, little is known about the requirements of the budding process. In this report, we have investigated the role of ATP in influenza virus budding by treating influenza virus infected Madin Darby canine kidney (MDCK) cells with a number of metabolic inhibitors. When WSN virus-infected MDCK cells were exposed to antimycin A, carbonyl cyanide m chlorophenylhydrazone, carbonyl cyanide p-trifluoromethoxy-phenylhydrazone, or oligomycin for a short time (15 min or 1 h) late in the infectious cycle, the rate of virus budding decreased. This inhibitory effect was reversible upon removal of the inhibitors. The role of ATP hydrolysis was analyzed by treating lysophosphatidylcholine (LPC)-permeabilized live filter-grown virus-infected MDCK cells with nonpermeable ATP analogues from the basal side and assaying virus budding from the apical side. In LPC-permeabilized cells, membrane-impermeable ATP analogues such as adenosine 5'-O-(3-thiotriphosphate) or 5' adenylylimidodiphosphate caused reduction of virus budding which could be partially restored by adding excess ATP. These data demonstrated that ATP hydrolysis and not just ATP binding was required for virus budding. However, inhibitors of ion channel (ATPases) and protein ubiquitinylation, which also required the ATP as energy source, did not affect influenza virus budding, suggesting that neither ion channel nor protein ubiquitinylation activity was involved in influenza virus budding. On the other hand, treatment with dimethyl sulfoxide (DMSO), which decreases membrane viscosity, reduced the rate of virus budding, demonstrating that the physical state of membrane viscosity and membrane fluidity had an important effect on virus budding. Data presented in the report indicate that influenza virus budding is an active ATP-dependent process and suggest that reduced virus budding by ATP depletion and DMSO treatment may be partly due to decreased membrane viscosity. PMID- 11883198 TI - French scallops: a new host for ostreid herpesvirus-1. AB - Sporadic high mortalities were reported among larval French scallops (Pecten maximus). Electron microscopy of moribund larvae revealed particles with the characteristics of a herpesvirus in association with cellular lesions. PCR and DNA sequencing showed that the virus is a variant of ostreid herpesvirus-1 that has already been described in clams and oysters. This is the first description of a herpesvirus infection of a scallop species. The virus was transmitted successfully from an extract of infected scallop larvae to uninfected scallop or oyster (Crassostrea gigas) larvae, demonstrating that it is able to infect both species. Detection of viral DNA in asymptomatic adult scallops by in situ hybridisation indicates that the herpesvirus may have been transmitted from adults to larvae. It is notable that, unlike most herpesviruses, this virus has a wide host range reflected by its ability to infect several species of marine bivalve. PMID- 11883199 TI - Effect of preexisting neutralizing antibodies on the anti-tumor immune response induced by chimeric human papillomavirus virus-like particle vaccines. AB - Chimeric human papillomavirus virus-like particles (HPV cVLPs) carrying HPV16 E7 protein are potent vaccines for inducing cell-mediated immunity (CMI) against HPV induced tumors in animal models. We tested the hypothesis that virion neutralizing antibodies generated during an initial vaccination might prevent effective boosting of CMI to the cVLPs. Mice with circulating HPV16-neutralizing antibodies, generated by direct immunization with wild-type VLPs or by passive transfer of hyperimmune anti-HPV16 VLP mouse sera, were subsequently vaccinated with HPV16 E7-containing cVLPs. Mice with preexisting neutralizing antibodies were not protected from HPV16 E7-positive TC-1 tumor challenge, compared to the protection seen in mice lacking these antibodies. Antibody-coated VLPs bound very inefficiently to receptor-positive cell lines, suggesting that one of the mechanisms of antibody interference is blocking of VLP binding to its receptor and thereby uptake of VLPs by antigen-presenting cells. Our results suggest that repetitive vaccination with a cVLP for induction of cellular immune responses to an incorporated antigen may be of limited effectiveness due to the presence of neutralizing antibodies against the capsid proteins induced after the first application. This limitation could potentially be overcome by boosting with cVLPs containing the same target antigen incorporated into other papillomavirus-type VLPs. PMID- 11883200 TI - [Vagina ecosystem with taking into consideration bacterial vaginosis in particular at pregnant women to be in miscarriage and premature delivery danger]. AB - The main tests of this writing was defining pregnant vagina ecosystem at hospitalized Clinic patients between 5th and 37th pregnant week. Patients was divided on two group: I-st with miscarriage danger, II-nd with premature delivery danger. Bacterial Vaginosis was recognized based into consideration Amsela criterion at 13.4% of examined patients, BV with mycosis coexisted at 3.8%, mycosis infection at 42.2% and trichomoniasis at 4.6% pregnant women. Influence occurred infection of Bacterial Vaginosis on premature delivery coming was also examined. Patients attended towards BV during the pregnant time, which gave preterm birth--21 (47.7%) compared with patients proper vagina ecosystem which also gave preterm labour premature--49 (36.6%). No radical, statistic comparison between those two groups was noticed. PMID- 11883201 TI - [Patient's preferences concerning the course of labor]. AB - OBJECTIVE: Much concern is being focused on the improvement of perinatal care standards in recent time. Not only the safety of woman and newborn, but also the comfort and individual preferences should to be considered. The aim of this study was to assess of expectations and requirements of the delivering women in relation to the course of labor and usage of the most common procedures in clinical practice. MATERIAL AND METHODS: 47 women who delivered in Obstetrical Word in Puck were questionnaire. Mean gestational age was 39 +/- 1.5 hbd. 47% of women were nulliparous, 53% were multiparous. The following variables were analyzed: the presence of medical staff and family at delivery, possibility of the delivery position choice, use of auxiliary devices, a friendly atmosphere during delivery, use of analgesia and labor induction, episiotomy and ante-partum preparation, cesarean section on request, attendance to labor school. Mann Whitney, Pearson and Yule tests were used for statistical analysis. RESULTS: 25% of women, mainly younger gravidae, attended the labor school. The midwife was considered the most important person at delivery. The presence of family member(s) was important for highly-educated women. The possibility to choose the delivery position and to walk during the 1st stage of labor was important for 73% of respondents. The majority of women who had attended the labor school avoided the horizontal position. Over 60% of patients accepted the usage of labor induction. A vast majority of women were against antepartum perineal shaving and episiotomy. Better-educated women preferred water delivery. 69% of the studied women would like to listen to the music at the delivery room. Cesarean section on request was supported by 11% of women. CONCLUSIONS: The tendency to promote modern delivery methods and active participation in labor leading is noticed. The significant influence of labor school on women's knowledge and their preferences was found. PMID- 11883202 TI - [Family labor from the father's point of view]. AB - Family labor becomes increasingly popular in our country. The aim of the study was answering the question: what the main motive for the men attending to labor was, whether fathers were satisfied of their participation and what the main reason of their satisfaction was. The study was performed in the district and local hospitals in south-eastern Poland during 7 months. 110 fathers were in the study group, 78 (70.9%) of urban population, and 32 (29.1%) of rural. The obtained results indicate firm acceptance of such a way of labor among majority of fathers. It was supported by the fact that almost all of them were satisfied of their participation in labor, and wished to attend the next one. The main reason for inclining them for the participation in labor was the granting of the partners' request, and internal willing. The source of satisfaction from sharing of the labor was providing the safety for the partner and setting up early contact with the baby. Significant influence of the labor schools on the feeling of readiness for delivery and the level of father's satisfaction were not evident. PMID- 11883203 TI - [Water birth in the parturients' estimation]. AB - OBJECTIVES: The purpose of the study was to characterise a group of women who decided for under water birth and to show an influence of warm water on their psychosomatic reactions. MATERIALS AND METHODS: The results of the inquiry conducted among 45 women bathed during first stage of normal labour and delivery under water were analysed. The demographic data, water birth knowledge and psychosomatic reactions were studied traditional. The age, education, obstetric history of the women was compared with a group of 45 women who gave birth in a way. RESULTS: The average age of women in labour under water was 25.6 years and no significant difference to control group (25 years) was found. The most common in our research group was secondary education (62%), after that elementary (20%) and university education (18%). Worse educated women were more rarely decided for water birth (20% in research group vs 41% in control; p < 0.05). A midwife was the most important source of information about warm tub bath during delivery, especially among worse educated women (67%). Concerning reactions after entering the pool, in 69% cases decrease of labour pain and in 64% decrease of spasm pains was observed. In 58% cases the time of delivery was advanced, only in 13% it lasted longer after going into the warm tub. Immersion in the pool was sensed in a positive way by all the parturients. The women described appeasement (78%), relaxation (67%), better opportunities for mid-spasm rest (67%). The water tub bath during delivery was estimated good by all of the women. 76% of the group gave 5 points in 1 to 5 scale. As many as 87% of women wish they born another baby in a water. CONCLUSIONS: A midwife has an essential role in information and making a decision of water birth. Entering the pool causes subjective decrease of labour pain and advance of delivery. Women very good estimate birth in water. PMID- 11883204 TI - [The influence of water immersion on the course of labor]. AB - OBJECTIVE: The aim of this study was to examine the influence of water immersion on the course of labor. MATERIAL AND METHODS: The study group consisted of 109 women, who have delivered in water in Obstetrical Ward in Puck from 1998 to 2000. 110 women composed control group. Mean patients' age in study and control group was respectively 26.40 +/- 4.33 and 26.72 +/- 5.82 years (ns). Gestational age was 40.69 +/- 5.91 and 39.71 +/- 2.03 weeks (ns). The duration of labor stages, time from membranes rupture to delivery, birthweight and newborns condition, frequency of episiotomy and perineum injuries as well as necessity of labor stimulants use were analyzed. Particular parameters were also assessed regarding to parity. The differences were determined using T-test. RESULTS: Mean duration of 1st labor stage was 319 min in study group and 375 min in control group (p < 0.02). The 2nd and 3rd labor stages did not differ significantly. II labor stage in nulliparous and I stage in multiparous were shorter in study group (respectively 34.41 vs. 45.5 min; p < 0.02 and 258.23 vs. 329.83 min; p < 0.02). The episiotomy was less frequent in study group (p < 0.01), whereas perineum injuries in control one (p < 0.05). Use of oxytocin was comparable between both groups. 97% of newborns from study group and 93% from control group, they were in good condition (ns). CONCLUSIONS: The profitable influence of water immersion to short 1st labor stage was noted. There were no differences in newborns' condition. The water birth is a safe method of labor in patients with physiological pregnancy. PMID- 11883205 TI - [Reproductive behavior after the flood disaster in Klodzko region--July 1997]. AB - RATIONALE: Flood disasters destroy environment of many people by causing physical as well as psychological harm. This may affect procreational behavior of the victims. DESIGN: This study examines reproductive behavior (number of live births) among the survivors from the flood disaster in Klodzko Region which took place in July 1997. The observation period was three years (1998-2000). MATERIALS AND METHODS: The harmed population consisted of 3986 subjects. The population from Klodzko Region which was exposed to flood disaster in 1997 consisted of 107,032 people. Among the injured population there were 1037 women in reproductive age. This population was studied. RESULTS: The results show significantly higher number of live births per 1000 and birth-rate in studied women as compared to the total population of a district. Psychological background of an observed phenomenon was discussed. PMID- 11883206 TI - [Violence towards pregnant women]. AB - The aim of this work was the evaluation of the scale of violence towards pregnant women in the westpomeranian province, the definition of the social-biological profile of women exposed to violence and social-biological profile of their partners. The evaluation of the influence of violence on pregnant women's ending term and the weight of the newborns. 481 women were enrolled and an anonymous study was used in the form of questionnaires. A questionnaire was a modified form of a query-sheet proposed by WHO. 25% of the enrolled women were exposed to physical and psychological (emotional) abuse, 7.1% to psychical violence, women and men exposed to violence in their childhood more often become violent in their adult life. Men that physically abuse pregnant women are often of primary school education, are unemployed, drink alcohol and smoke. Physical abuse by a partner during pregnancy usually experience women with primary school education, who drink and smoke. Violence during pregnancy is usually associated with premature delivery as well as low birth weight of the newborns. PMID- 11883207 TI - [Birth weight and state of dentition in children with transitional dentition with regard to oral hygiene]. AB - The aim of the present study was to determine the dependency between the state of dentition and the birth weight of the pupils of the third grade of primary schools in Lublin. The subjects of the examination were 150 aged 9 year and 8 months +/- 3.9 months (76 girls and 74 boys). Clinical and questionnaire tests and a statistical analysis were carried out. The obtained results and literature review seem to allow for the following conclusions: there is a relationship between the sum of dmfs and DMFs indexes and the advancement of the carious process and OHI--S--with increment of birth weight and decrement of OHI--S index the sum of dmfs and DMFs indexes decreased; there was no correlation between disturbances of teeth mineralization and the birth weight. PMID- 11883208 TI - [Cadmium concentration in uterine myomas, myometrium and peripheral blood from women living in Lower Silesia]. AB - The author has studied the cadmium concentration in uterine myomas, myometrial tissue and peripheral blood in 72 women living in Lower Silesia. The highest cadmium concentration was observed in myometrial tissue, lower in uterine myomas and the lowest in the peripheral blood. The differences among the tissues were statistically significant. There were no significant differences in the cadmium concentration concerning the women's age. There was significant positive correlation in the cadmium concentration between the myomas and the myometrium. PMID- 11883209 TI - [The analysis of sodium, potassium and calcium concentration in myomas, myometrium and peripheral blood]. AB - We have examined the concentration of sodium and potassium in myomas, myometrium and peripheral blood of 55 women and in 69 women calcium was examined. The levels of these elements were correlated to the phase of the menstrual cycle. The sodium concentration is similar in the examined tissues in both phases. The potassium concentration is significantly higher in myomas and myometrium in comparison with the peripheral blood in both phases of the menstrual cycle. The calcium concentration is significantly lower in myomas and myometrium during the proliferation and significantly lower in the myomas comparing to the myometrium and peripheral blood in the secretion. Concerning the differences between two phases of the cycle we observed that the level of the potassium is significantly lower in the myomas in the proliferative phase but the the level of the sodium is higher in the secretory phase. PMID- 11883210 TI - [The influence of paternal lymphocytes immunization on percentage of peripheral blood CD16+/CD56+ cells in women with primary recurrent spontaneous abortion]. AB - OBJECTIVES: Paternal lymphocytes immunization has been proposed for several years as an efficient treatment for unexplained Recurrent Spontaneous Abortion (RSA), however precise mechanism that underline the benefits of this immunotherapy is still unclear. DESIGN: The aim was to study the influence of paternal lymphocytes immunization on percentage of peripheral blood NK cells (CD16+/CD56+) in women with primary RSA of unknown etiology. MATERIALS AND METHODS: 48 patients with history of 3-5 (mean 3.3 +/- 0.7) consecutive primary RSA of unknown etiology were selected for the study. Immunotherapy with paternal lymphocytes, isolated from 100 ml of peripheral blood, was performed twice prior conception with a 4 week interval. The percentage of NK cells (CD16+/CD56+) was estimated using standard flow-cytometric immunofluorescent techniques for whole blood with one step monoclonal anti-CD16/CD56 antibodies. Statistical analysis was performed with Wilcoxon test and the p value less than 0.05 was accepted as statistically significant. RESULTS: It was found that paternal lymphocytes immunization significantly decreases the percentage of NK cells in peripheral blood in women with RSA (23.9 +/- 8.5 vs. 16.2 +/- 7.0; p = 0.009). CONCLUSIONS: The data of the present studies suggest that paternal lymphocytes immunization modulate immunity in women with primary unexplained RSA significantly decreasing the percentage of NK cells (CD16+/CD56+) in peripheral blood. PMID- 11883211 TI - [Clinical features of missed abortion]. AB - RATIONALE: There is no general agreement concerning the definition of missed abortion which remains one of the most commonly encountered pregnancy complication of an extremely variable clinical picture. AIM OF THIS STUDY: The analysis of the clinical symptoms of missed abortion. SETTING: Academic Medical Center. MATERIALS AND METHODS: Consecutive 50 women with non-viable pregnancy from 7 to 22 weeks, diagnosed by clinical examination, ultrasonography and serum beta-HCG evaluation were studied. RESULTS: In 92% of the missed abortion cases a vaginal spotting was observed before the diagnosis was established. The average period of the estimated retention of the products of conception was 2.8 weeks. In only one patients this period exceeded 8 weeks. The reverse correlation was established between the gravidity of a patient and the period of asymptomatic retention of the non-viable conception products in uterus. The obtained results confirm that the onset of vaginal bleeding does not reflect the moment of embryonal/fetal death. PMID- 11883212 TI - [The antioxidant-prooxidant balance in pregnancy complicated by spontaneous abortion]. AB - The purpose of this study was to characterize prooxidant-antioxidant balance in patients with different clinical types of spontaneous abortion. Sixty previously untreated, pregnant women aged 16-38 years were included into this study. The gestational age ranged from 8 to 12 weeks. Patients were divided into three groups of 20 subjects each. The first group consisted of patients with inevitable abortion. The second group included women with threatened abortion in whom pregnancy was maintained until term. The control group consisted of women with uncomplicated pregnancy. In all the patients venous blood was collected immediately after the diagnosis was settled and before the treatment was commenced. Two biochemical markers of oxidative stress were quantified, namely: lipid peroxides (LP) using spectrophotometric TBARS method and total antioxidant status (TAS) by means of the commercially available kits (TAS: Randox Laboratories Ltd., U.K.). The obtained results indicate that spontaneous abortion is accompanied by a profound disruption of the prooxidant-antioxidant homeostasis towards oxidative stress. PMID- 11883213 TI - [Laminin levels in patients with ectopic pregnancy]. AB - Serum laminin (sL--by means of ELISA method) was evaluated in 17 patients with tubal pregnancy (TP) and 25 healthy control subjects. In patients with ruptured TP sL was significantly elevated as compared to both the controls and the patients with unruptured TP (p < 0.001). The obtained results suggest the usefulness of sL measurements in clinical management of patients with symptoms of ectopic pregnancy. PMID- 11883214 TI - [Markers of coagulation and fibrinolysis in peripheral blood and in blood collected pouch of Douglas in patients with ectopic pregnancy]. AB - In cases of ectopic pregnancy a blood collected in a pouch of Douglas is a nonclotting one. It contains no fibrinogen, prothrombin, factor V and VII, plasminogen and monomers of fibrin, however concentration of D-dimers is very high. Is this solely an effect of independently occurring fibrinolysis or does coagulation accompany this process? The definite answer can not be given. Coagulation and fibrinolysis markers obtained at the same time from peripheral veins were normal. PMID- 11883215 TI - [The postnatal condition evaluation of preterm infants]. AB - OBJECTIVE: The aim of the study was to compare general postnatal condition of preterm infants delivered by caesarean section or born vaginally. MATERIAL AND METHODS: The study group consisted of 605 premature newborns delivered in Obstetrics and Gynaecology Clinic of Medical University of Warsaw in 1995-2000. The newborns were divided into two groups. There were 280 prematures delivered by caesarean section in the first group and 325 ones were born vaginally in the second group. RESULTS: The first day mortality rate in infants born vaginally was 6.8% comparing with 2.8% in ones delivered by caesarean section. CONCLUSIONS: In the study group the percentage of neonatal severe condition was similar in both groups but mortality rate in the first day of life was slightly higher in infants born vaginally. PMID- 11883216 TI - [Management and outcome of preterm labor in department of obstetrics and pathology of pregnancy school of medicine in Lublin]. AB - OBJECTIVES: We present data of three years experience of management and outcome of preterm labour. MATERIALS AND METHODS: The study comprised 335 pregnant women with preterm labour. We analyzed the pharmacological therapy and way of labour. There were two groups of patients: I group--180 patients who had cesarean delivery, II group--155 patients who had vaginal delivery. RESULTS: It has been found 7.54% more cesarean delivery than vaginal delivery of preterm labour. PMID- 11883217 TI - [Application of 2% clindamycin cream in the treatment of bacterial vaginosis and valuation of methylcellulose gel containing the complex of Chitosan F and PVP k 90 with lactic acid as carrier for intravaginally adhbited medicines in the cases of pregnancies with the symptoms of preterm delivery]. AB - OBJECTIVES: There are many reports informing about the connection between BV and the increased risk of preterm delivery. The reason of self-concession and reversion of BV after having executed an efficient treatment has not yet been properly explained. DESIGN: The aim of this work was the clinical valuation of the 2% Clindamycin cream in the treatment of BV and of the methylcellulose gel containing the complex of Chitosan F and PVP K-90 with lactic acid as a carrier for intravaginally adhbited medicines in the cases of pregnancies with the symptoms of a preterm delivery. MATERIAL AND METHODS: The research comprised 145 pregnant between 24-34 week of pregnancy, hospitalised because of the symptoms of a preterm menace delivery. In the case of the detection of BV, a 10-day therapy using intravaginal cream containing 2% Clindamycin was executed. In the cases not qualified as BV, the methylcellulose gel containing the complex of Chitosan F and PVP K-90 with lactic acid has been intravaginally adhibited for 10 days. CONCLUSIONS: 1. Application of 2% Clindamycin cream is an efficient method of the treatment of bacterial vaginosis. 2. Methylcellulose gel containing lactic acid combined with the complex of Chitosan F and PVP K-90 allows a persistent maintenance of the correct pH of vagina. 3. Methylcellulose gel, because of its physico-chemical properties similar to physiological mucus, is a universal carrier for intravaginally adhibited medicines. PMID- 11883218 TI - [Analysis of the postnatal condition and mortality of neonates with extremely low birth weight]. AB - OBJECTIVE: The aim of this study was to analyze the postnatal condition and mortality of neonates with extremely low birth weight. MATERIALS AND METHODS: The study group consisted of 35 neonates delivered in the 1st Dept of Ob/Gyn. Medical University of Warsaw in the period of 1996-2000. The group was divided into two classes depending on the birth weight. There were newborns weighted 500-750 g in the first class and 751-1000 g in the second one. Newborns condition in the 1st minute of life was assessed with Apgar score. The rate of mortality up to the 7th day of life as well as the causes of deaths was analyzed. CONCLUSIONS: Postnatal mortality rate of premature newborns extremely low birth weight is still very high, especially in case of newborns below 750 g. Respiratory distress syndrome and intracranial hemorrhage are the most common causes of demise of those newborns. PMID- 11883219 TI - [Short-term and long-term fetal heart rate variability after amnioinfusion treatment of oligohydramnios complicated pregnancy]. AB - Results of computerised analysis of cardiotocograms obtained in the group of 21 pregnancies complicated by idiopathic oligohydramnios are presented in the study. Amnioinfusion procedures were administered serially in local anesthesia with ultrasound and colour Doppler control on the base of oligohydramnios criteria by Phelan. The analysis was based on KOMPOR software created by ITAM Zabrze based on PC computer connected to Hewlett-Packard Series 50A cardiotocograph. Significant short-term variability increase just after amnioinfusion procedure from 5.55 ms to 8.24 ms and after 24 hours up to 7.25 ms was found, while significant long term variability values changes were not observed. PMID- 11883220 TI - [Emergency cerclage with concomitant amniocentesis as a method of treatment of membranes prolapse--case report]. AB - Emergency cerclage was performed on 27 years old patient with membrane prolapse in 21 weeks' gestation. Ultrasonographically guided amniocentesis was performed intraoperatively in order to decrease intraamniotic pressure. Aggressive prophylactic management (wide spectrum antibiotics and tocolysis-nifedipine) resulted in pregnancy prolongation up to 30 week of gestation. At that time healthy premature newborn (1520 g, Apgar score 7 in 1st minute) was delivered by caesarean section. The mother was discharged from the hospital on the 4th postoperative day, whereas the child after 5 weeks weighted 2130 g. PMID- 11883221 TI - [Amniocentesis and mother's bladder overfilling in operative treatment of advanced cervical incompetence]. AB - We describe two cases treated for cervical incompetence with prolapsed fetal membranes with emergency cerclage. Two methods of membranes reducing were used: transabdominal amniocentesis and bladder overfilling. We recommend these procedures in patients with advanced cervical incompetence when the membrane reduction by Trendelenburg position or pharmacological tocolysis were ineffective. PMID- 11883222 TI - [Preterm deliveries of extremely low birth babies--analysis of the course of pregnancy, labour and neonatal outcome]. AB - OBJECTIVE: Preterm delivery of baby with extremely low birth weight (ELBW) is an important problem in contemporary perinatology and a major reason of perinatal mortality. A great development of neonatal intensive care that has occurred over the last few years has resulted in the decrease of perinatal mortality rate. The aim of the study is to analyse the course of pregnancy, labour and neonatal outcome in the group of patients who delivered ELBW children. MATERIALS AND METHODS: In 1st Obstetric and Gynaecology Department Clinic of Medical University of Warsaw, 6982 deliveries were conducted from 1996 to 2000. Among them there were 589 preterm ones. As a result 44 women delivered ELBW children. These 44 deliveries were divided into three groups according to the reasons: Idiopathic preterm contractility-24 patients PROM with intrauterine infection or threatening infection-14 Induction of delivery because of lethal foetal defects and direct foetal distress-6. RESULTS: The most common reasons for deliveries of ELBW children were hypertension and ascending infections. CONCLUSIONS: A great number of mothers who delivered ELBW children had low socioeconomic status. In most cases pregnancy was unplanned and body mass index of the mothers was below 19. Almost all these newborns were severely depressed and delivery was often complicated. It is important to conduct labour in a careful way to avoid tissue injuries. PMID- 11883223 TI - [Ultrasound fetal biometry and birth weight prognosis with the use of Aoki method and classical regression analysis]. AB - We have attempted to estimate prognostic value of neonatal birth weight prediction with the use of fetal sonographic data: biparietal diameter, femur length, transverse and longitudinal abdominal diameter. Clinical and ultrasound data from 265 pregnant women with singleton gestation who delivered within 7 days from the last ultrasound scan were analyzed. For the fetal weight estimation we have calculated fetal abdominal area in the equation proposed by Aoki in 1990. Predicted and observed neonatal birth weight centiles and coefficients of determination were calculated with the use of regression analysis. Prediction method of neonatal weight with Aoki method explained 70% of variability in our population. In contrast, regression analysis on ultrasound data explained 99% of observer variability in neonatal birth weight. We conclude that own method of birth weight prediction could be of value, however it should be tested in a new prospectively examined population. PMID- 11883224 TI - [Sonographic evaluation of cervical length and cervical canal width in 24th week of twin pregnancy in prediction of preterm delivery]. AB - OBJECTIVE: Sonographic evaluation of the cervical length and cervical canal width in 24th week of twin pregnancy in prediction of preterm delivery. MATERIALS AND METHODS: 43 primiparous patients with twin pregnancy. Group I (n = 16, 37.2%) with spontaneous delivery before 36 of gestation; group II (n = 27, 62.8%) delivering in term. Sonographic measurements of the cervical length and cervical canal width were done in 24th week of gestation. RESULTS: Mean values of cervical length in group I were: 25.6 mm (+/- 3.7 mm); and in group II--32.5 mm (+/- 6.0 mm). Corresponding values of canal width were: 14.6 mm (+/- 11.8 mm) and 6.2 mm (+/- 3.0 mm). CONCLUSIONS: We suggest, that cervical length is more valuable parameter in predicting risk of preterm delivery in twin pregnancy. PMID- 11883225 TI - [Ultrasonographic assessment of cervical length and measurement of fetal fibronectin level in cervical secretion of pregnant women in prophylaxis of premature deliveries]. AB - OBJECTIVE: The aim of this study was to assess the usefulness of the assessment of cervical length and and measurement of fetal fibronectin level in cervical secretion of pregnant women in prophylaxis of premature deliveries. MATERIAL AND METHODS: 78 pregnant women hospitalized in Department of Fertility and Obstetrics, University School of Medicine Wroclaw. They were divided into III groups: Group I-13 pregnant women who had premature delivery and the time between examination and delivery was no longer than 24 hours. Group II-20 women who had premature delivery, and the time between examination and delivery was longer than 24 hours. Group III-pregnant who delivered at term (control group). Between the 25th and the 34th week of pregnancy presence of fetal fibronectin in cervical secretion and ultrasonographic assessment of cervical length were done. RESULTS: Significantly higher percentage of women with presence of fibronectin in cervical secretion and significantly shorter length of cervical length was stated in 13 pregnant women who delivered prematurely, between the 28th and 35th week of pregnancy comparing with other groups. In group II pregnant women delivered between the 28th and 35th week of pregnancy, and the time between examination and delivery was from 3 days to 4 weeks. In this group in 75% examined women fibronectin was present in cervical secretion. Significant was that cervical length in 14 of this group of women (70%) was no longer than 20 mm and in the rest was between 20-30 mm, moreover in no cases was longer than 30 mm. In group III only in 7% women presence of fibronectin in cervical secretion was stated and only in two cases (4%) cervical length was shorter than 20 mm, both of them delivered in 38th week of pregnancy. CONCLUSIONS: 1. Examination of fetal fibronectin in cervical secretion allows precisely estimate the risk of premature delivery. 2. Cervical length shorter then 20 mm collerates with the presence of fetal fibronectin in cervical secretion and with increased risk of premature delivery. PMID- 11883226 TI - [Levels of fetal and plasma fibronectin in patients with cervical incompetence]. AB - In 45 gravidae (below 24 weeks) fetal fibronectin (fF) was measured in cervico vaginal discharge and plasma fibronectin (pF) was evaluated in venous blood. Twenty women had cervical incompetence (CI) diagnosed and 25 healthy gravidae served as a control. The fF level in patients with CI (0.073 +/- 0.014 mg/ml) was significantly (p < 0.01) higher than that found in the control group (0.058 +/- 0.011 mg/ml). The same was true for pF (p < 0.001). A diagnostic value of fibronectin levels in patients suspected of cervical incompetence is suggested. PMID- 11883227 TI - [Concentration of the cellular fibronectin in amniotic fluid and plasma in the peripartum period]. AB - OBJECTIVES: Numerous publications recommending the determination of fibronectin (FN) and fetal fibronectin (fFN) during pregnancy and delivery have appeared. The aim of this study was to analyse the level of cellular fibronectin concentration in amniotic fluid and blood plasma of patients between 38 to 42 pregnancy weeks. MATERIAL AND METHODS: The research comprised 79 women in 38-42 weeks of pregnancy. The FN concentration was determined in amniotic fluid and plasma by the solid phase immunoenzymatic method (ELISA) with the use of mice monoclonal antibody (TaKaRa FN 30-8). RESULTS: The FN concentration in amniotic fluid ranged from 0.001 to 0.18 mg/ml whereas in plasma from 0.15 to 3.0 mg/ml. In the cases of determination of FN in amniotic fluid the most of the samples (64 of 83; 77%) were enclosed between 0.025 and 0.08 mg/ml. In plasma 70% of the results (57 of 81) ranged from 0.25 to 0.8 mg/ml. CONCLUSIONS: In 38-40 weeks of pregnancy cellular FN concentration increases whereas in 41-42 weeks it decreases gradually. No differences were observed in the fibronectin concentration level in the blood plasma and amniotic fluid of pregnant women as compared to parturients. PMID- 11883228 TI - [The importance of fibronectin in amniotic fluid in anticipation of premature labor]. AB - We analysed 86 pregnant women where we estimated the fibronectin level in specimens of amniotic fluid. During carrying out the experiment we noted that fibronectin is present in amniotic fluid and can be identified in a quantity mode. We have proved dependence between fibronectin level in amniotic fluid and the period of time from collecting the sample, up to the delivery. Fibronectin level in amniotic fluid doesn't depend on pregnancy duration, preterm rupture of amniotic membranes. PMID- 11883229 TI - [Maternal serum cytokines in labor, pregnancy and chorioamnionitis]. AB - The purpose of our study was to compare maternal serum levels of interleukin-6, interleukin-8, tumor necrosis factor-alpha and interferon-gamma in gravidities, during spontaneous term and preterm labor and their relation to histologic chorioamnionitis. METHODS: We investigated 61 women: 10 in preterm labor, 36 in term labor and 15 healthy pregnant nonlabouring controls. Venous bloods for cytokines determinations were obtained during the first stage of labor and during routine screening tests. Titers of cytokines were measured by means of ELISA technique. All births after preterm deliveries were examined to establish histologic chorioamnionitis. RESULTS: Serum levels of IL-6 and IL-8 were significantly elevated both in term (mean: IL-6: 17.5 +/- 58 pg/ml; IL-8: 148 +/- 215 pg/ml) and preterm labor (IL-6: 23 +/- 44 pg/ml; IL-8: 332 +/- 389 pg/ml) when compared to nonlabouring gravidities (IL-6: 5 +/- 7 pg/ml; IL-8: 14 +/- 11 pg/ml). IL-6 and IL-8 titers were statistically similar in term and preterm labors and in patients with and without histologic chorioamnionitis. TNF-alpha and IFN-gamma were not statistically analyzed because only a few patients had detectable serum levels of these cytokines. CONCLUSION: Serum levels of IL-6 and IL-8 in both: term and preterm labor are elevated in comparison to nonlabouring gravidities. The elevated levels of these cytokines are not connected with coexisting chorioamnionitis. PMID- 11883230 TI - [The relationship between the levels of proinflammatory cytokines, and AFI value in pregnancies complicated by premature rupture of membranes]. AB - OBJECTIVE: The aim of the study was to test the relationship between maternal serum proinflammatory cytokines level and AFI (amniotic fluid index) in pregnancies complicated by premature rupture of membranes (PROM). MATERIALS AND METHODS: The maternal serum levels of IL-6, IL-1 beta and TNF-alfa were assessed in patients with PROM between 24-34 weeks of gestation (n = 45) by means of commercially available ELISA assays. The patients were divided in two groups according to AFI values: < 50 (n = 25) mm and < or = 50 mm (n = 20). Cytokine concentrations were compared between groups. RESULTS: The median concentrations of proinflammatory cytokines in maternal serum were: IL-6--5.74 pg/ml (range 3.24 3111 pg/ml), IL-1 beta--0.76 pg/ml (range 0.001-3.16 pg/ml), TNF-alfa--1332.46 pg/ml (range 2.13-1969.68 pg/ml). Compared to patients with AFI values > or = 50 mm, the group of patients with AFI < 50 mm had significantly higher concentration of IL-6 (6.61 pg/ml vs. 4.66 pg/ml; p = 0.002). No significant differences in IL 1 beta and TNF-alfa levels have been found between groups. The significant correlation have been observed between maternal serum level of IL-6 and AFI values (R = -0.47, p = 0.003), but not of IL-1 beta and TNF-alfa. CONCLUSION: The assessment of AFI values in pregnancies complicated by premature rupture of membranes seems to be the valuable method of early diagnosis in cases of intrauterine infection. PMID- 11883231 TI - [The condition of premature newborns of patients hospitalized due to premature rupture of fetal membranes (PROM)]. AB - OBJECTIVES OF THE STUDY: The aim of the study was to compare the premature newborns' condition of patients hospitalized due to PROM with the group of premature newborns of patients with amniorrhexis at the time of labor. MATERIAL AND METHODS: The studied group consisted of 74 newborns of 70 gravidas hospitalized in the Department of Pathological Pregnancy and the control group of 77 newborns of 70 patients. On analysing the investigated materials special attention was paid to: the time of PROM, the time and the way of pregnancy, the age of the, obstetrical history, the symptoms of intrauterine infection. In assessment of the newborns' condition the following factors were taken into consideration: characteristics of infection, Apgar scores, birth weight, respiratory disorders and other newborns' diseases. RESULTS: Theoretic risk of sepsis was higher in study than in control group (p = 0.00). The sepsis occurrence was comparable in both groups (7 vs 8). The infection occurred more frequently in study group. CONCLUSIONS: 1. The newborn condition depends on the week of pregnancy termination, with better prognosis for longer pregnancies and with no differences between both groups 2. In spite of statistically important risk of sepsis occurrence in study group, the real occurrence was comparable in both groups 3. There is a statistically important difference between CRP and 5' Apgar score in study group. 4. Percentiles of birthweight and birthlength appropriate for gestational age were comparable between both groups. PMID- 11883232 TI - [Influence of premature rupture of fetal membranes on kynurenic acid concentration in umbilical cord blood]. AB - The aim of the study was to evaluate the influence of premature rupture of fetal membranes on kynurenic acid (KYNA) levels in venous and arterial umbilical cord blood of neonates. Statistically significant higher concentration of KYNA in umbilical arterial and venous blood in neonates of mothers with PROM lasting longer than 11 hours was observed. Increase of KYNA concentration in blood of such babies may be one of markers of developing infection. PMID- 11883233 TI - [The usefulness of the intrapartum fetal pulse oximetry in anticipating the neonatal outcome]. AB - OBJECTIVE: The aim of the study was to evaluate the usefulness of the intrapartum fetal pulse oximetry in anticipating the neonatal outcome. MATERIALS AND METHODS: The saturation of the fetal blood (SpO2) was measured during labor with non invasive pulse oximeter designed for fetal application. The average, minimum and maximum SpO2 were evaluated separately for the first and the second stage of labor. The average SpO2 of the fetus was compared to neonatal condition assessed by umbilical vein pH, pO2 and pCO2 and according to Apgar score. RESULTS: Twenty patients have been monitored with fetal pulse oximetry. All those patients had normal vaginal delivery. During the first stage of labor, the average fetal SpO2 was 51.94 +/- 8.03%, the minimum SpO2 was 38.35 +/- 9.15%, and the maximum SpO2 was 63.35 +/- 7.75%; in the second stage of labor average fetal SpO2 was 43.82 +/ 7.16%, minimum SpO2 was 34.35 +/- 7.79% and the maximum SpO2 was 50.94 +/- 8.37%. A significant decrease in fetal average and maximum SpO2 occurred from stage I to stage II of labor (average SpO2: 51.94 +/- 8.03% vs. 43.82 +/- 7.16%, p = 0.0002; maximum SpO2: 63.35 +/- 7.75% vs. 50.94 +/- 8.37%, p < 0.00001). The significant correlation between the average SpO2 during the first stage of labor and umbilical vein pH (R = 0.60, p = 0.02) and pO2 (R = 0.54, p = 0.04) was found. No relationship between fetal SpO2 in the first and second stage of labor and Apgar score was observed. CONCLUSIONS: 1. The second stage of labor results in significant decrease in fetal SpO2. 2. The fetal SpO2 > 30% in the first and second stage of labor is related to good neonatal outcome. 3. The fetal SpO2 assessment in first stage of labor seems to be important in newborn's acidosis and hypoxemia predicting. PMID- 11883234 TI - [The management of labor overweight women with intrapartum fetal oxygen saturation monitoring]. AB - OBJECTIVE: Overweight is one of the most common problems that occurs in pregnancy. The aim of this study was to evaluate the usefulness of fetal pulse oximetry during labour and to compare fetal oxygen saturation between cases with pregravid overweight and normal weight. MATERIAL AND METHODS: Fetal oxygen saturation was continuously recorded with use of Nellcor N-400 fetal pulse oximeter in 20 cases of pregravid overweight and 30 control cases of normal weight. Distribution of fetal oxygen saturation values during 4 periods of labour was analyzed and compared between the examined groups together with neonatal umbilical artery pH values, Apgar score, birth weight and percentage of ceasarian sections performed. RESULTS: We noticed statistically important differences in fetal oxygen saturation between analyzed groups. Mean oxygen saturation value was lower in the overweight group, at the end of first stages of the labour (47% vs 52%) and at the second stages (42% vs 46%) We noticed differences in birth weight too. No significant differences in neonatal umbilical artery pH, ceasarian sections, newborns mean 1 minute Apgar score were observed between analyzed groups. CONCLUSION: Fetal pulse oximetry is a a useful method for intensive surveillance of the fetus at risk of hypoxemia during the labour. A lower fetal oxygen saturation value during labor by pregravid overweight were observed. PMID- 11883235 TI - [Overweight and obesity as the risk factor in perinatology]. AB - Overweight and obesity have become a frequent phenomenon among pregnant women during last thirty years. They result in increased morbidity rates of different chronic, health- or even life-threatening diseases. Among different perinatal complications associated with obesity the most important are: hypertension, diabetes, varices, cholecystolithiasis, prolonged pregnancy, intrauterine growth retardation. Increased rates of operative deliveries, intrapartal and postpartal infections, thrombotic complications, anaemia, urinary infections and lactation disorders can be observed. PMID- 11883236 TI - [Evaluation of the platelet angiotensin receptor in pregnancy complicated by preeclampsia]. AB - Preeclampsia is a major complication during human pregnancy. It results from a breakdown in the balance between the vasoconstrictor and vasodilator substances. Angiotensin II is a potent vasoconstrictor, which participates in the regulation of blood pressure and may be involved in the control of vascular tone. OBJECTIVE: The purpose of this study was to determine the platelet angiotensin II receptor number and angiotensin II level in pregnancies complicated by preeclampsia. Preeclampsia was defined according to American College of Obstetricians and Gynecologists (ACOG) classification. PATIENTS AND METHODS: Ligand binding techniques were used for the examination of platelet angiotensin II binding sites in the third trimester pregnant women. The study was carried out in 13 patients with singleton pregnancy complicated by preeclampsia. A control group consisted of 17 healthy normotensive patients with singleton uncomplicated pregnancy and normal laboratory tests. All studied patients were nonsmokers. RESULTS AND CONCLUSIONS: There were no statistically significant differences in patient profiles between groups including gravidity, parity, maternal age, gestational age and height. Maternal weight, BMI and systolic, diastolic blood pressure and mean arterial blood pressure (MAP) were higher in the study group in comparison with the control group. Our study revealed elevated platelet angiotensin II receptor number and decreased maternal angiotensin II level in singleton pregnancies complicated by preeclampsia. There were no correlations between platelet angiotensin II receptor number and plasma angiotensin II level in the studied subjects. Our results are in accord with other published data and point out to the significant role of renin-angiotensin-aldosterone system and angiotensin II receptors in the pathogenesis of preeclampsia. PMID- 11883237 TI - [Assessment of selected parameters of kidney activity among women with pregnancy and delivery complicated by EPH-gestosis]. AB - This paper estimates the excretory kidneys activity in women with pregnancy complicated by EPH-gestosis and in women with a physiological pregnancy by denoting albuminuria, beta-2-microglobulinuria and beta-2-microglobulin in blood serum. It was claimed that the excretion of albumins with urine increases considerably in women with a pregnancy complicated by EPH-gestosis before the delivery in comparison with women with a pregnancy of a physiological course. The increase of albumins excretion with urine in a labour in two groups of tested pregnant women was also noticed. However, the increase in excretion of albumins with urine during delivery was much bigger in the group of women with pregnancy complicated by EPH-gestosis in comparison to the group of women with a physiological course of pregnancy. The albumins size before delivery as well as during the next 48 hours after the delivery did not differ statistically in both groups of tested women. Beta-2-microglobulin concentration in blood serum was bigger in the group of pregnant women with EPH-gestosis than women with a physiological pregnancy during the whole period of research. Bigger excretion of beta-2-microglobulin with urine was noticed among women with pregnancy complicated by EPH-gestosis before delivery, during delivery and during the next 48 hours after the delivery in relation to the group of women with a physiological pregnancy. PMID- 11883238 TI - [Oxidative stress in pregnancies complicated by gestational hypertension (GH), effect of oxidized serum lipoproteins (O-LDL)--the role of antio-LDL antibodies]. AB - OBJECTIVE: To explain correlations between metabolism of serous oxidized lipoproteins, their influence on placental function in pregnancies complicated by GH and intrauterine asphyxia. MATERIALS AND METHODS: The group of 233 pregnant women characterised by gestational age between 32-40 weeks were divided into two sub-groups: the study group of 105 women with GH and the control group of 128 healthy pregnant women. In all the subjects doppler flows in umbilical and middle brain artery as well as the serum levels of anti o-LDL antibodies (oLAB) were measured. RESULTS: In the study group mean PI values in middle brain artery equal 1.62 +/- 0.41 while in the control group 0.91 +/- 0.16 (p < 0.01), with oLAB values 1133 +/- 24 for the study group and 619 +/- 15 for the controls. Having analysed vascular flows in umbilical artery, mean PI values of 1.41 +/- 0.36 for the study group and 0.63 +/- 0.12 for the control group were obtained, (p < 0.01). The levels of oLAB in these groups were as follows: 1019 +/- 18 for the study group and 600 +/- 12 for the controls. CONCLUSIONS: 1. In pregnancies complicated by GH an intensified oxidative stress was observed as determined by increased levels of anti o-LDL antibodies (oLAB). 2. Pulsatility indices in umbilical and middle brain artery exhibited enhanced values in pregnancies complicated by GH. PMID- 11883239 TI - [Assessment of lipid peroxidation intensification in normal and preeclamptic placentas]. AB - OBJECTIVE: Free radical induced lipid peroxidation (LP) in the placenta has been suggested as a possible pathogenetic factor of preeclampsia (PE). DESIGN: The aim of the study was to assess LP intensification by the measurement of lipid peroxidation products (LPP) content in placentas from normal and preeclamptic pregnancies. MATERIALS AND METHODS: The investigations comprised placentas obtained immediately after delivery from 24 normal pregnancies [group K], 26 pregnancies complicated by severe PE without intrauterine growth restriction (IUGR) [group PE] and 23 pregnancies complicated by severe PE and IUGR [group PEI]. LPP content was measured by the quantitative determination of thiobarbituric acid reactive substances (TBA-RS) amounts in studied placentas. Used TBA test was calibrated with malondialdehyde (MDA) and results were expressed as MDA equivalent in nmol/mg protein. Comparative analysis was performed using U Mann-Whitney and median tests. RESULTS: Mean placental level of LPP (MLPP) in the group PE-2.45 +/- 0.39 (M +/- SD) was significantly higher (p < 0.001) as compared to MLPP in the group K (1.58 +/- 0.24). MLPP in the PEI group (2.81 +/- 0.65) was higher (p < 0.001) than MLPP in the group K as well as MLPP in the group PE but statistical significance of the latter difference was lower (p = 0.032). CONCLUSIONS: The intensification of LP in placentas from pregnancies complicated by severe PE is IUGR dependent and higher than in placentas from normal pregnancies. Obtained results may indicate that higher degree of LP intensification in preeclamptic placentas may be involved in PE pathogenesis. PMID- 11883240 TI - [Parameters of peripheral whole blood neutrophil activity in women with pregnancy induced hypertension]. AB - OBJECTIVES: Pregnancy induced hypertension is believed to be a state of neutrophil overactivity, however all previous studies were done on isolated cells. DESIGN: To study neutrophil activity in whole blood of PIH women. MATERIALS AND METHODS: Neutrophil activity was estimated without isolation, in peripheral whole blood of 23 PIH women and 26 normal pregnant controls. The chemiluminescence test was performed without any stimulation and upon stimulation of neutrophils with fMLP, OZ and PMA, before and after pre-activation with TNF alpha. The results were corrected according to the haemoglobin concentration and (%) of neutrophils. The percentage of whole blood neutrophils indicating expression of selectins CD18, CD11b, integrin CD62L and mean fluorescence intensity (MFI) of these molecules were studied on flow-cytometry. RESULTS: The study revealed that neutrophil chemiluminescence was not significantly higher in PIH women and after correction coefficient used it was even lower in PIH patients. TNF-alpha preactivation had no influence on chemiluminescence results. Expression of CD11b expressed as MFI value was significantly increased while that of CD62L, expressed as (%) of positive cells and MFI value--decreased in PIH patients. CONCLUSIONS: Changes of neutrophil CD11b and CD62 expression indicate increased activity of these cells in PIH women, however low production of reactive oxygen species estimated by corrected chemiluminescence test especially after TNF-alpha pre-activation, indicates that this form of their reactivity is rather "exhausted" during PIH. PMID- 11883241 TI - [Catalase activity in normal and preeclamptic placentas]. AB - OBJECTIVE: The decrease of placental catalase (CAT) activity may lead to an increase of placental amounts of reactive oxygen species and can contribute to preeclampsia pathogenesis. DESIGN: The aim of the study was to determine CAT activity in placentas from normal and preeclamptic pregnancies (with and without intrauterine growth restriction--IUGR). MATERIALS AND METHODS: The investigations comprised placentas obtained immediately after delivery from 22 normal pregnancies (group K), 26 pregnancies complicated by severe preeclampsia-PE without IUGR (group PE) and 23 pregnancies complicated by severe PE and IUGR (group PEI). The activity of CAT was determined using a spectrophotometric method and expressed as IU/mg protein. Comparative analysis was performed using U Mann Whitney test. RESULTS: Mean activity of CAT (MCAT) in the PEI group--0.38 +/- 0.14 (M +/- SD), was significantly lower (p < 0.001) as compared to MCAT in the group K (0.55 +/- 0.16). MCAT in the PE group (0.48 +/- 0.14) was lower than MCAT in the group K, but this difference was not statistically significant (p > 0.05). MCAT in the group PEI was significantly lower (p = 0.026) as compared to MCAT in the PE group. CONCLUSIONS: The activity of CAT is decreased in placentas from pregnancies complicated by severe PE and IUGR. Obtained results may indicate that the decrease of placental CAT activity may be involved in pathogenesis of IUGR in preeclamptic pregnancies. PMID- 11883242 TI - [Decidual lymphocyte subsets in pregnant women]. AB - OBJECTIVES: Materno-foetal immunological reactions in decidua are probably one of the most important elements in pathogenesis of preeclampsia. DESIGN: To compare lymphocyte subsets isolated from decidua of preeclamptic pregnant women with lymphocyte subsets isolated from healthy pregnant women. MATERIALS AND METHODS: Preeclampsia (PE) was defined according to USA National Health Institute criteria. The study group consisted of 21 women with PE and 11 women with physiological pregnancy. All pregnancies were finished with elective cesarean sections. Exclusion criteria were: uterine contractions, infection, chorinamnionitis, diabetes mellitus and therapy with steroids less than 7 days before blood sampling. Decidual tissue was obtained by curettage of the uterine cavity. The fragments of decidua were separated from clotted blood and placed in sterile tubes with 5 ml of isotonic solution (PBS). Then the decidual tissue was mechanically fragmented, homogenized and rinsed in PBS. Routine immunofluorescent marking techniques with monoclonal antibodies were performed. Analysis was done with FACSCalibur flow-cytometer with 488 nm argon laser using CellQuest programme. The following lymphocyte subsets were estimated: CD3, CD19, CD4, CD8, CD4/CD29, CD8/CD28, CD4/CD45RA, CD4/CD45RO, CD56/CD16, CD69. The results were described as percentage of lymphocytes positive for above surface molecules. Statistical analysis was performed using t-Student and U-Mann-Whitney tests. The work was sponsored by KBN 4 P05E 118 15 grant. RESULTS: Decidua of pregnant PE women contains significantly increased percentage of CD3-/ CD56 + 16+, CD8+/CD28+ cells and decreased percentage of CD3+, CD19+, CD4+/CD29+ and CD4+/CD45RA+ compared to decidua of healthy pregnant controls. CONCLUSIONS: These changes suggest that deficiency of suppressor activity as well as aberrant immunoregulation exists in decidual tissue of PE women. PMID- 11883243 TI - [DNA adducts level in normal and preeclamptic placentas]. AB - OBJECTIVE: An increase in lipid peroxidation intensification in preeclamptic placentas leads to an increased level of lipid peroxidation products and increased reactive oxygen species activity which can be associated with increased activation of chemicals to electrophilic species that bind covalently to DNA and form adducts. DESIGN: The aim of the study was the determination of DNA adducts (A-DNA) in placentas from normal and preeclamptic pregnancies. MATERIALS AND METHODS: The investigations comprised placentas obtained immediately after delivery from 21 normal pregnancies [group K], 24 pregnancies complicated by severe preeclampsia-PE without intrauterine growth restriction (IUGR [group PE] and 21 pregnancies complicated by severe PE and IUGR [group PEI]. DNA adducts were determined using nuclease P1 digestion enhancement version of the 32P postlabeling method. The results were expressed in numbers of DNA adducts per 10(8) nucleotides. Comparative analysis was performed using ANO-VA and median tests. RESULTS: Mean level of A-DNA (MA-DNA) in the group PE--1.39 +/- 1.21 (M +/ SD) was similar (p = 0.57) to MA-DNA in group K (1.16 +/- 1.03). However MA-DNA in the PEI group (1.93 +/- 1.28) was significantly higher (p = 0.045) than MA-DNA in the group K as well as MA-DNA in the group PE (p = 0.025). MA-DNA level in all studied preeclamptic placentas (groups PE + PEI) was 1.65 +/- 1.26 and was similar (p = 0.152) to revealed in group K. CONCLUSIONS: The level of DNA adducts in placentas from pregnancies complicated by severe preeclampsia and IUGR is higher than in placentas from pregnancies complicated by severe preeclampsia without IUGR and higher than in placentas from normal pregnancies. PMID- 11883244 TI - [Arterial hypertension during pregnancy complicated by type-1 diabetes--clinical aspects]. AB - In the group of 289 pregnant diabetic women hospitalised and followed-up between 1991-2000 in the Maternal-Fetal Medicine Dept., Research Institute Polish Mothers Memorial Hospital, 44 patients were diagnosed with hypertension arterialis (15.2%), significantly more frequently in women with long lasting diabetes complicated by angiopathy and whose who trend to be obese. Metabolic control did not differ in the group with hypertension and without. In the group of pregnant women with hypertension following symptoms occurred significantly more frequently: proteinuria (29.5%), pyelonephritis (11.4%), anaemia (25%) and the risk of premature delivery (25%). Hypertension arterialis shortened significantly the duration of pregnancy (34.7 weeks of gestation vs. 37.3) and affected the obstetrical outcome such as:-low birth weight and longer time of newborn hospitalisation. PMID- 11883245 TI - [Non omnis moriar. In memory of professor Radzislaw Sikorski]. PMID- 11883246 TI - [Zoonoses in reproductive medicine]. AB - After reviewing the importance of three zoonoses, namely toxoplasmosis, listeriosis and brucellosis, Authors focus on their experience in prophylactic treatment of the acquired toxoplasmosis in 72 gravidae with high risk pregnancy and the history of toxoplasmosis. Listeriosis and brucellosis currently seem not to bear a clinical significance as perinatal risk factors. Toxoplasmosis, however, should be taken into account a cases of unexplained pregnancy loss. The Authors' experience suggests the positive effect of the combined prophylactis antitoxoplasmic treatment in the selected cases. PMID- 11883247 TI - The significance of ante- and perinatal periods for formation of risk of sudden infant death syndrome. AB - The results of the research, which are carried out according to two international programs on studying and prevention of the sudden infant death syndrome (SIDS), are submitted in the article. The features of ante- and perinatal periods, characteristics of obstetric history of the mothers of deceased babies are analyzed, the relative risk of SIDS development is estimated at the influence of a complex of factors. High parity, young age of mother at the time of delivery, poor antenatal care, preterm delivery, intrauterine fetal growth retardation, fast delivery, alcohol and coffee abuse by a mother, smoking during pregnancy, early change of the child's feeding from breast to bottle are referred to the most significant risk factors of SIDS stipulated by the unfavorable course of pregnancy and delivery. The practical recommendations directed on elimination of the risk factors of SIDS in ante- and perinatal periods are offered. PMID- 11883248 TI - [Epidemiology of the pregnant women risk groups according to Troszynski's criterion with special regard to risk factors of potentially environmental character]. AB - THE AIM: To evaluate the occurrence of specific increased risk groups of pregnancies against a background of the environment contamination in Rybnik. Environmental pregnancy risk factors were particularly taken into consideration. METHODS: Pregnant women giving birth in the Obstetrical Department in Rybnik in the years 1986-2000. Were study subject. The occurrence of factors dependent on the environment was the object of the inquiry. RESULTS AND CONCLUSIONS: The concentration of chemical substances in Rybnik in the years 1988-2000 revealed a declining tendency. Uncomplicated pregnancies were noticed in 50% of the cases. There was no evident correlation between professional activity and the occurrence of increased risk groups. PMID- 11883249 TI - [Cotinine levels in serum and colostrum of smoking puerperae]. AB - In 50 women (25 smokers and 25 non-smokers), cotinine, a relatively stable metabolite of nicotine, was measured in the 3rd postpartum day in blood serum and colostrum by means of enzyme-enhanced chemiluminescence. Non-smokers had cotinine levels in serum and colostrum below 10 ng/ml. Colostrum cotinine content in smokers amounted to approximately 50% of the respective serum level and both these values were strongly positively correlated (p < 0.0014). Cotinine level in early breast-milk did not correlate significantly with the number of cigarettes smoked daily by a mother. Newborns breast-fed by smoking women are exposed to the tobacco smoke compounds not only by an environmental ("passive") smoking but also by the ingestion of nicotine metabolites present in mother's milk. PMID- 11883250 TI - [Ultrasonographic diagnosis of fetal congenital abnormalities]. AB - Retrospective analysis of ultrasonographic examinations in 81 women who delivered newborns with congenital malformations was performed. Totally 147 anatomical defects in 83 newborns were found. 85 of 147 abnormalities (57.8%) were detected prenatally, most commonly (79.6%) in the third-trimester. All cases of coverings defects, as well as, all cases of anencephaly were diagnosed with ultrasonography. Malformations of urinary tract in 94.4%, central nervous system in 82.9%, osteoarticular system in 30%, heart defects in 27.8% and face abnormalities in 21.4% were already sonographically visualized before birth. PMID- 11883251 TI - [Teratogenic action of antiepileptic drug: 2-propylpentanoic acid-effects on rat and hamster embryos cultured in vitro]. AB - Valproic acid (2-propylpentanoic acid) has become accepted as one of the most important antiepileptic drug. Although clinically effective, this drug remains controversial as suspected of being a human teratogen. The aim of our study was to test the embryotoxic potential of valproic acid using whole embryo culture assay. For that purpose rat and hamster embryos were explanted on gestation day 9 and 7 respectively, and cultured for 48 and 24 hours. During the culture period embryos were exposed to valproic acid (VPA) at a concentrations of: 10, 50 and 100 mg/ml. In our study this drug exhibited dose-related embryotoxic and teratogenic effects in rat and hamster embryos. Analysis of the results indicated that hamster embryos were more susceptible to VPA than rat embryos. PMID- 11883252 TI - [Three dimensional sonography in nuchal translucency measurements between 10th and 14th weeks of gestation]. AB - Our objective was to determine the practical usefulness of 3D transvaginal sonography in nuchal translucency (NT) measurements between 10th and 14th week of gestation. Thirty-six women (18 primiparas) were studied with the use of Kretz Voluson 530 and 730 scanners (Kretztechnik, Zipf, Austria). Two different volume scans were performed in all subjects according to the new 4 steps method proposed by Chung et al (2000). Nuchal translucency of the first and second scan were compared. In all but one (2.7%) cases good quality images were obtained. Mean value of NT of the first and second scan were 1.463 +/- 0.551 mm (SD) and 1.475 +/- 0.556 mm (SD), respectively. The differences as compared by two tailed t-test for paired measurements were not significant (p = 0.668). There was a significant correlation between first and second measurement (r = 0.96), also the interoperator variability was good. Our preliminary study indicates that 3D sonography is a useful tool in nuchal translucency measurements at the end of the first trimester of pregnancy. The results, however, need to be prospectively tested in larger population. PMID- 11883253 TI - [Arrhythmias in perinatal period and neonatal outcome--analysis of own experiences]. AB - OBJECTIVE: The paper presents review of different kinds of fetal arrhythmias, diagnosed in our centres. DESIGN: The aim of our study was to analyse results of echocardiographic examinations of 141 fetuses with diagnosed arrhythmias referred for fetal cardiac examination in years VI. 1996-V. 2001 and neonatal outcome. RESULTS: Premature contractions were recognised in 113 (80%) fetuses, tachyarrhythmias in 7 (5%) cases and complete atrioventricular block in 5 (4%). Heart defects were diagnosed in three cases. Three fetuses developed congestive heart failure (2x SVT, 1x HLHS + block a-v III*). From all group 6 newborns died. CONCLUSION: In case of fetal arrhythmias echocardiography should be widely applied in order to evaluate the kind of arrhythmia, sufficiency of circulation and coexistence of heart defects or functional abnormalities with arrhythmia in fetal circulation. PMID- 11883254 TI - [Analysis of stillbirths in Rybnik in the years 1991-2000]. AB - 153 cases of stillbirths, which took place in Gynecological & Obstetrical Department in Rybnik by 1991-2000 were analyzed. In one third of the cases women did not receive prenatal care. Most stillbirths were noticed in a birth weight range between 501-1000 g. The majority of parturients were unemployed and multiparous ones. PMID- 11883255 TI - [Semen analysis results and air pollution in the group of men from infertile couples in the Lower Silesia in 1977-2000]. AB - OBJECTIVES: To evaluate the relation among the concentration of selected air pollution and seminal parameters, examined from 1977 to 2000. DESIGN: Semen analysis and air pollution results were retrospectively evaluated. MATERIALS AND METHODS: We have analysed semiograms from 1363 men from infertile couples inhabiting Lower Silesia. Seminal volume, sperm concentration, percentage of pathologic sperms were measured in all men. Estimation of mean seminal volume, total sperm number, sperm motility, and percentage of pathologic sperms per year was performed. Average values for NO2, CO, SO2 and dust concentrations during the study were taken from official sources. RESULTS AND CONCLUSIONS: We showed the statistically significant increase in the percentage of pathological sperms (R2 = 0.9, p < 0.05), the slight increase in the semen volume(R2 = 0.4, p < 0.05). The total sperm count in the semen samples revealed a slight increase (R2 = 0.25, p < 0.05). The sperm concentrations and the percentage of motile sperms remained relatively stable and relatively constant. Statistically important decrease in NO2 (R2 = 0.85, p < 0.05), SO2 (R2 = 0.96, p < 0.05) and dust concentration (R2 = 0.89, p < 0.05) and no change in CO concentration was revealed. There is no correlation among concentrations of dust, NO2, SO2, CO and the increase in percentage of pathologic sperms. PMID- 11883256 TI - [Administration of natural anthocyanins derived from chokeberry (aronia melanocarpa) extract in the treatment of oligospermia in males with enhanced autoantibodies to oxidized low density lipoproteins (oLAB). The impact on fructose levels]. AB - PURPOSE: To determinate the influence of anthocyanins from chokeberry on the generation of serous autoantibodies to oxidized low density lipoproteins (oLAB) and fructose levels in semen of males with oligospermia. MATERIAL AND METHODS: Study group consisted of 38 males with oligospermia of who were randomly divided into 2 subgroups: in 22 subjects anthocyanins were administered, while 16 men were on placebo; control group consisted of 25 healthy male volunteers. We examined the level of oxidative stress measured by serum concentration of oLAB, together with fructose levels in semen. RESULTS: In the anthocyanin group oLAB titres decreased from 1103 +/- 34 mU/ml before treatment to 742 +/- 24 mU/ml (1st month) and 724 +/- 23 mU/ml (2nd month), p < 0.01. In the placebo group oLAB titres were as follows--1094 +/- 21 mU/ml before treatment, 1114 +/- 36 mU/ml (1st month) and 1117 +/- 33 mU/ml (2nd month), p > 0.05; oLAB tires in the control group: 601 +/- 40 mU/ml before treatment, 609 +/- 38 mU/ml (1st month) and 609 +/- 38 mU/ml (2nd month), p > 0.05. In the anthocyanin group fructose levels in semen increased from 850 +/- 42 mg/ml before treatment to 1121 +/- 26* mg/ml (1st month) and 1230 +/- 27* mg/ml (2nd month), p < 0.05. In the placebo group fructose levels were as follows--832 +/- 36 mg/ml before treatment, 845 +/- 33 mg/ml (1st month) and 841 +/- 35 mg/ml (2nd month), p > 0.05; fructose levels in the control group: 1376 +/- 40 mg/ml before treatment, 1376 +/- 40 mg/ml (1st month) and 1388 +/- 37 mg/ml (2nd month), p > 0.05. CONCLUSIONS: 1. Our results indicate that natural antioxidants (anthocyanins) can be useful in controlling of oxidative stress in males with oligospermia. 2. Anthocyanins from aronia melanocarpa induce enhanced levels of fructose in semen of males with oligospermia. PMID- 11883257 TI - [The influence of smoking on the estradiol level in the preovulatory follicular fluid]. AB - The aim of the present study was to test the relationship between the objectively measured intrafollicular exposure to the tobacco smoke compounds and the estradiol level in the preovulatory follicular fluid. In 30 IVF-ET patients (14 smokers and 16 non-smokers) levels of cotinine (C-FF) and 17 beta-estradiol (E2 FF) were quantified in the follicular fluid by means of radioimmunoassay. In smokers (C-FF from 62.21 to 595.00 ng/ml) the mean E2-FF was 85.07 nmol/l, while in the non-smokers (C-FF from 1.20 to 15.62 ng/ml) the respective value amounted to 109.19 nmol/l. The difference was statistically significant (p = 0.005). The obtained results support the growing body of evidence that tobacco smoking negatively influences endocrine function of the Graafian follicle. PMID- 11883258 TI - [Effect of vitamin/mineral supplementation on calcium, magnesium and iron levels in amniotic fluid and serum taken from women during first half of pregnancy]. AB - The levels of calcium, magnesium and iron in amniotic fluid and serum taken from women during first half of pregnancy were compared. The relationship between these levels and vitamin-mineral supplementation was also estimated. There was similar calcium and magnesium concentration in the serum and amniotic fluid. The concentration of iron in serum was four times as large as in amniotic fluid. Any influence of supplementation on concentration of studied minerals was noted. PMID- 11883259 TI - [The influence of demographic and behavioral factors on the risk of human papillomavirus infection]. AB - OBJECTIVES: The aim of this work was the assessment of correlation between different types of HPV and chosen factors: the number of pregnancies, childbirths and smoking habits. MATERIALS AND METHODS: Patients were examined in the Gynaecology Clinic, Silesian Medical Academy in Zabrze from 1998 until 2000. They were either hospital patients or women subjected to routine cytological examination. The smears were simultaneously collected for both cytooncologic examination and PCR identification of HPV viruses. Cytological smears were classified according to the Bethesda system. RESULTS: A relationship between the presence of HPV and number of pregnancies or childbirths was confirmed as well as the relationship between smoking and the presence of HPV genome. The higher number of pregnancies or childbirths the higher frequency of HPV presence is. HPV presence was more frequent in the group of smoking women. CONCLUSIONS: A relationship between the presence of HPV and the number of past childbirth or pregnancies was found. The statistically significant correlation between smoking and the risk of HPV infection was also confirmed. PMID- 11883260 TI - [Determination of antioxidative-peroxidative balance in the cord blood of newborns delivered to mothers with diabetes type G1]. AB - The aim of the study was to evaluate antioxidant system activity and lipid peroxidations products as markers of oxidative stress in the cord blood of newborns delivered to mothers with diabetes type G1 in comparison to healthy pregnant women. MATERIALS AND METHODS: We examined 10 healthy and 10 diabetic mothers' newborns. Total antioxidant status (TAS) and thiobarbituric acid reactive substances (TBARS) were measured using spectrophotometry. We compared also birth weight, gestational age, Apgar score and cord blood pH between groups. RESULTS: There were no significant differences in birth weight, gestational age, Apgar score and pH value between the groups. We found two cases of congenital malformations and two severe hyperbilirubinemia in diabetic mothers' newborns. We found also decreased level of TAS and higher level of TBARS in the cord blood of newborns delivered to mothers with diabetes type G1 in comparison to the control group. CONCLUSIONS: Our results suggest that maternal diabetes G1 during pregnancy induces oxidative stress in the newborn. PMID- 11883262 TI - Levels of mannose binding lectin in early pregnancy complicated by diabetes mellitus type 1--preliminary report. AB - The complement system plays an important role as a product of innate and acquired immune reaction. It can be activated via three different routes: the classical pathway, the alternative pathway and the lectin pathway. MBL (mannose-binding lectin) is considered to be a pathogen recognising receptor (PRR), an important factor in recognising pathogen associated molecular pattern (PAMP). The aim of study was to evaluate MBL in early pregnancy of diabetic mothers. Higher values of MBL were observed in diabetic non-pregnant women compared to healthy non pregnant. Subjects early pregnancy seems to decrease MBL values in both diabetic and healthy pregnant women. PMID- 11883261 TI - [Effect of intensive prenatal and diabetic care on newborn of diabetic mothers- 14 years of observation]. AB - Modern management of pregnancies complicated by maternal diabetes mellitus has in view delivery at term an healthy child with correct body mass and in good metabolic condition. THE AIM: Of our work was estimation of influence of intensive prenatal and diabetical cares on frequency of occurrence of failures and complications at newborns of mothers with pregestational diabetes mellitus delivered in period from 1987 to 2000. Study include 228 of newborns of diabetic mothers of class B-RF according to White, born in our hospital between 25 and 40 weeks of pregnancy. In aim of estimation of percentage of perinatal mortality, congenital malformations, prematurity, hyper- and hypotrophy in examined population of new-born children of mothers with PGDM, material was subdivided into 3 groups in 4 and 5-years intervals: group I (n = 96) 1987-1990, group II (n = 72) 1991-1995, group III (n = 60) 1996-2000r. RESULTS: Systematical decrease of percentage of perinatal mortality in examined groups from 7.3% to 5.0% was ascertained. There was a significant association between decreasing of number of congenital malformations in investigated new-born population from 12.5% to 5.0% and 2,5-times increasing of percentage of women with PGDM managed by combined team of diabetologist and obstetricians before conception. We observed an essential decreasing of percentage of prematurity in investigated material from 51.6% to 20.0%. Percentage of births of new-born children with macrosomy on space analysed of years was comparable. (13.7%, 14.1% and 15.0%) in spite of increasing of percentage of childbirths at term from to 80%. We did not ascertain increasing of percentage of incidents of IUGR. The percentage of IUGR in investigated groups carried out 5.3%, 5.6% and 5.0% in spite 2-times increasing of percentage of births of children of mothers with diabetes in class R and RF according to the White classification. CONCLUSIONS: Well controlled diabetes, intensive management of diabetic women by combined team of diabetologists and obstetricians on space 14 of years of activity of our Department due to decreasing of percentage of: perinatal mortality from 7.3% to 5.0%, congenital malformations from 12.5% to 5.0% and premature deliveries from 51.6% to 20.0% in population of new-born children of mothers with PGDM. PMID- 11883263 TI - [Pregnancy complicated by a neoplasmatic disease]. AB - The Author reviews the problem of neoplasmatic complications of pregnancy focusing mainly on genital tumors such as vulvar, vaginal, cervical, uterine, tubal and ovarian neoplasms as well as on other pelvic tumors originating from the bladder and bowel. Non-genital neoplasms such as breast cancer, Hodgkin's disease, non-Hodgkin lymphoma, leukemia, melanoma and thyroid cancer are also addressed. Literature data on the incidence of tumors in pregnant women are reviewed and the therapeutic options considering both the mother and the fetus are discussed. Management recommendations are given in cases when following the treatment a patient becomes pregnant again. PMID- 11883264 TI - [Difficulties in differential diagnosis of adnexal masses during pregnancy: the role of greyscale and color doppler sonography]. AB - We have attempted to determine the accuracy of greyscale and color Doppler ultrasound in the differentiation of adnexal masses in pregnancy. The studied group included 2245 pregnant women from low risk population. Following criteria were evaluated: maximal diameter and volume of the tumor, echogenicity, presence of septa and papillary projections in grey scale sonography. Color Doppler analysis included blood vessel presence and arrangement and blood flow characteristics with the use of pulsatility (PI), resistive (RI) and systolic/diastolic (S/D) indices. Preoperative CA-125 serum levels were available in 11 patients. In 66 (2.94%) patients adnexal tumors were detected during routine ultrasound scan at the end of the first trimester. Twenty-seven masses (1.2%) persisted beyond 16 weeks of gestation and were subsequently surgically removed. Pathological diagnosis confirmed 19 serous cystadenomas, 4 endometriomas and 2 dermoids, one pedunculated myoma and one fibrothecoma. Mean size of the tumors was 79 Jmm (range: 43-245 mm), mean volume 166. lml (range: 30-1320 ml). Doppler indices values presented as mean, SD and range were as follows: PI = 1.26 +/- 0.71 (range: 0.57-3.84); RI = 0.61 +/- 0.15 (range: 0.33-0.89) and S/D = 2.62 +/- 0.98 (range: 1.17-4.91). Median serum concentration of CA-125 was 17 IU/ml (range: 8.4-1247 IU/ml). Only 3 of these women had elevated (> 35 IU/ml) levels: 2 endometriomas (344 IU/ml and 1247 IU/ml) and one myoma (37 IU/ml), respectively. Based on the sonographic findings two solid tumors were incorrectly classified as probably malignant (fibrothecoma and subserous myoma). Negative predictive value of ultrasound diagnosis in the studied population was therefore 92.6% (25 of 27). We conclude that although prenatal sonography has the potential to correctly classify most of adnexal masses, caution in risk assessment is needed especially when persistent solid tumor is found. PMID- 11883265 TI - [Pregnancy delivery and newborn in female kidney recipients]. AB - OBJECTIVE: During the past few years a great development of clinical transplantology has taken place. Organ transplantation permits the best and quickest biological, clinical and social rehabilitation of patients. The subsidence of hormonal disturbances, which are responsible for irregular ovarian cycles and ovulation, has been observed among women with stable graft function. Such patients often become capable of conceiving and possessing offspring. With an increasing frequency of such occurrences, the pregnancy and delivery in the female transplant recipients is now becoming one of the major problem of contemporary perinatology. The aim of this study was to summarize our experience with the pregnancy, delivery and the state of newborn in female kidney transplant recipients. MATERIALS AND METHODS: 25 female kidney-transplant recipients who became pregnant were observed during pregnancy. The age of the patient was between 23 and 38 years. 32 pregnancies were followed up. The duration and complications of pregnancy, mode of delivery and newborns condition were analyzed. RESULTS: Miscarriages were observed in 47% of cases. Among patients who got birth, premature delivery was observed in 53%. Two stillbirths took place. 91% neonates were born in good condition. Preterm delivery occurred in 53%. Average birth weight was lower then observed at healthy pregnant women. No congenital defects were observed. CONCLUSIONS: In kidney transplant recipients who became pregnant the increased incidence of spontaneous abortion and preterm delivery was observed. Newborns delivered by those patients had decreased birth weight. No congenital defects were noted among those babies. PMID- 11883266 TI - [The course of pregnancy, delivery and puerperium in women with varices and thrombophlebitis of lower extremities, after application of low molecular weight heparins]. AB - OBJECTIVES: Estimation of the long term prophylactic or therapeutic application of low molecular weight heparin (LMWH) on the platelets count, and incidence bleedings during pregnancy, delivery and puerperium in the women with varices of lower extremities and past thrombophlebitis of lower extremities. MATERIAL AND METHODS: 5212 pregnant, women in labour and in puerperium divided into 4 groups.; 142 women with varices and thrombophlebitis of lower extremities (group I); 10 with past thrombophlebitis of lower extremities (group II); 15 with thrombophlebitis in current pregnancy; 5045 without vascular complications (group IV--control). In group I during pregnancy compression therapy was applied (stockings) and low molecular weight heparins (LMWH) in course of puerperium. In group II during the 1st trimester of pregnancy and in labour the same heparin doses were administered, while the doses were increased in the 2nd and 3rd trimester. In group III, when thrombophlebitis was stated non-steroid anti inflammatory drugs and LMWH were administered. In all cases treated with heparin both number of platelets and incidents of bleedings from genitourinary tract were observed. Presence of embolic complications was also noted. RESULTS: No cases of decrease platelets number or bleedings from genitourinary tract were observed in group I-III during administering of LMWH. In women in group II where prophylactic with LMWH was applied no incidences of recurrent thrombophlebitis during pregnancy and puerperium were observed. In group I-III all newborns were born in good condition and no complications were observed. Average blood loss during both labour and cesarean section, among women in group I-III was not significantly different comparing with control group. No incidences of pulmonary artery embolism or decrease number of platelets were observed. CONCLUSIONS: 1. The long term prophylactic or therapeutical administration of LMWH in the women with varices of lower extremities or thrombophlebitis has no influence on the platelets count and incidence of bleedings from genitourinary tract during pregnancy or increase of blood loss during labour and puerperium. 2. In the women with past thrombophlebitis of the lower extremities after application of LMWH during pregnancy there were no recurrence observed. PMID- 11883267 TI - [Pregnancy and delivery in women with thrombocytopenia]. AB - Maternal immune thrombocytopenia is a frequent finding in pregnancy and is most commonly asymptomatic and clinically benign. 8 pregnant women with thrombocytopenia between January 1995 and December 2000, in Clinic of Obstetrics and Gynecology in Katowice-Ligota, were evaluated. 7 patients had idiopathic thrombocytopenia purpura and one (31 years old) had cirrhosis, splenomegaly, thrombocytopenia and elevated liver enzymes. 6 cases was steroid-treated with initial platelet count 4,000 per cubic millimeter to 115,000 per cubic millimeter after treatment. Alloimmune thrombocytopenia is a serious fetal disorder, resulting from platelet-antigen incompatibility between the mother and fetus. Among all newborns only one had thrombocytopenia, platelet count 45,000 per cubic millimeter, without neonatal intracranial hemorrhage. 6 women had cesarean section and 2 patients delivered naturally. PMID- 11883268 TI - [Comparison of vaginal misoprostol and oxytocin for labor induction in post-term pregnancy]. AB - OBJECTIVES: To compare labour induction intervals between vaginal misoprostol and intravenous oxytocin as well as side effects of induction in post term pregnancies with intact membranes. METHODS: One hundred women were retrospectively selected to two groups treated with vaginal misoprostol 50 micrograms every 12 hours as needed to maximum 150 micrograms and treated with intravenous oxytocin. The primary outcome measure was time from induction to vaginal delivery. Statistical analysis was performed by t-Student test. RESULTS: Maternal age, parity, gestation were similar. There was a statistically important difference in labour induction intervals between the two groups. The mean time +/ SD to vaginal delivery in misoprostol group was 20.6 +/- 15.2 hours compared with 11.23 +/- 7.4 hours with oxytocin (p = 0.0396). Induction of labour failed in 12% and 32% in misoprostol and oxytocin treated group. Pethidine consumption in oxytocin treated group was higher (41 mg vs 89 mg, p = 0.04). Episodes of vomiting were more frequent in misoprostol treated group (22% vs 6%). There were no episodes of uterine hyperstimulation in both groups. CONCLUSIONS: Oxytocin stimulation resulted in a shorter induction to delivery interval. In misoprostol group induction failed in only 12% whereas in oxytocin group in 32%. There were no serious side effects in both groups. In misoprostol treated group patients required less analgetics then in oxytocin treated group. PMID- 11883269 TI - Influence of antisperm antibodies on T cell adhesion to extracellular matrix proteins in women with a history of unexplained infertility. AB - beta 1 integrins, receptors responsible for the binding of extracellular matrix proteins, mediate activation, adhesion, migration and differentiation in different tissue compartments. It has been recently demonstrated that integrins are distributed on sperm surface and on T cells, as well. The aim of this study was to establish influence of antisperm antibodies on T cell adhesion to extracellular matrix proteins in women with a history of unexplained infertility. PMID- 11883270 TI - [Transvaginal ultrasonography and diagnostic hysteroscopy in infertile women]. AB - Sensibility and specificity of transvaginal ultrasonography and diagnostic hysteroscopy were compared in 211 infertile women. Examinations were performed between 1996-2000 in I-st Department of Obstetrics and Gynecology Warsaw Medical University--Poland. Comparing results we concluded high compatibility of transvaginal ultrasound and hysteroscopic examinations. In the future--as we suppose--the number of diagnostic hysteroscopies in infertile women will decrease and will be at least partially replaced by transvaginal ultrasound as less invasive and less expensive method. PMID- 11883271 TI - [Peritoneal fluid and plasma 4-hydroxynonenal and malonyldialdehyde concentrations in infertile women]. AB - OBJECTIVES: To investigate the possible role of oxidative stress in the pathogenesis of female infertility, we examined the peritoneal fluid (PF) and plasma lipid peroxides levels in infertile women. MATERIALS AND METHODS: 4 hydroxynonenal (4-HNE) and malonyldialdehyde (MDA) concentrations were measured in peritoneal fluid and plasma samples obtained from: 10 infertile women with minimal and mild endometriosis, 24 patients with unexplained infertility, 11 women with PCOS and 14 patients with tubal infertility. RESULTS: PF lipid peroxides level was significantly higher in women with idiopathic infertility compared to other groups. Plasma 4-HNE and MDA concentrations did not differ significantly between the infertile groups. No significant difference or relationship was observed between peritoneal fluid and plasma lipid peroxides levels. CONCLUSIONS: Increased free radicals activity in the peritoneal fluid environment may be the factor responsible for some cases of "unexplained infertility". PMID- 11883272 TI - [Semen parameters in secondary infertile couples]. AB - OBJECTIVE: The aim of this study was to evaluate sperm values in the group of secondary infertile couples. MATERIAL AND METHODS: The material of the study included 35 couples with abortion in anamnesis, 12 couples with delivery in past and 11 couples conceive and deliver during studies. Sperm analysis was performed according to the WHO guidelines. RESULTS: Men from couples with abortion in past had significantly higher sperm concentration compare to couples with delivery in past. In other parameters there was no significant differences observed. CONCLUSION: Sperm concentration can be important factor in secondary infertility. PMID- 11883273 TI - [Levels of proinflammatory cytokines (Il-1 alpha, Il-6, TNF-alpha) in the semen plasma of male partners of infertile couples]. AB - In semen plasma of 46 male partners of infertile couples the levels of proinflammatory cytokines such as IL-1 alpha, IL-6, TNF-alpha were quantified. The studied group was divided according to the number of leucocytes in an ejaculate into two subgroups (> 10(6)/ml and below 10(6)/ml). The levels of the studied cytokines were significantly elevated (p < 0.05) in patients with higher leucocyte count in the ejaculate. Testing the immunologic markers may be helpful in diagnostic process considering male partners of infertile couples. PMID- 11883274 TI - [Biophysical characteristics of cervical mucus after stimulation of ovulation by clomiphene citrate and HMG]. AB - OBJECTIVES: Characteristics of cervical mucus during monofollicular stimulation of ovulation by clomiphene citrate and by clomiphene citrate with HMG was evaluated. MATERIALS AND METHODS: 226 women of couples treated for male factor infertility or idiopathic infertility were studied. Favourable parameters of cervical mucus in women treated by clomiphene citrate with HMG were found although they were worse than observed in spontaneous ovulatory cycle. RESULTS AND CONCLUSIONS: The E2/follicle index was used for evaluating efficiency of stimulation. It was the highest during cycles were CC and HMG were used, although they did not reflect the risk of hyperstimulation. Authors discuss the importance of their findings in cases when techniques of ART can not be applied. PMID- 11883275 TI - [IL-6 and TNF-alpha levels in oviductal epithelial cells culture after escherichia coli LPS stimulation]. AB - The aim of the study was to determine wether Gram-negative bacterial cell membrane lipopolysaccharides (endotoxine) can change the IL-6 and TNF-alpha cytokine concentration synthetized by fallopian tube endothelial cells. For the study 5 normal fallopian tubes from females at their reproductive age who underwent total hysterectomy due to uterine myoma were used. The fallopian tubes specimens (endothelial tissue) 2 mm2 fixed in 0.5 ml DMEM/HAM F-12 (GibcoBRL) solution with 15% FCS, Gentamycin, Fungizone, ITS (GibcoBRL) were incubated at 37 degrees C with 5% CO2. The explants were stimulated by Escherichia coli lipopolysaccharide (LPS) at ascending concentrations 1 ng/mL, 10 ng/mL and 100 ng LPS/mL incubation media. Tissue specimen incubated in a media without LPS were used for control test. IL-6 activity in the supernatants were determined by Van Sinc method, TNF-alpha activity were determined against WEHI-164.13. cells according to Espevik and Nisser-Mayer. The presence of TNF-alpha and IL-6 were confirmed in all the supernatants of the incubated fallopian tubes explants. The LPS stimulation caused a concentration increase in both cytokines. The maximum cytokine concentration level was observed in the incubation stimulated at 1 and 10 ng LPS/mL incubation media. The use of the highest LPS concentration retarded the cytokine production. CONCLUSION: The TNF-alpha and IL-6 cytokines dose dependent production is caused by the fallopian tubes LPS stimulation. PMID- 11883276 TI - [Changes in carbohydrate metabolism of PCOS patients in cycle stimulated by rFSH]. AB - 24 patients with clomiphene citrate-resistant PCOS were enrolled in the study. We used low-dose step up protocol of recombinant FSH administration. Ovulation induction by rFSH injection was commenced on 3 day of cycle, at a dose of 50 IU for 7 days initially. The dose was continued at 50 IU, and ultrasonographic scans were performed daily until a dominant follicle had developed to a size of at least 18-20 mm, at which point 5,000 IU of hCG was administrated. If no response was noted after 7 days, the dose was increased by 50 IU for a further 7 days. Insulin and glucose serum levels were determined 7 times: in days 3, 5, 8 of a cycle and later when diameter of ovarian follicle was 10, 12, 16 mm and in the day of hCG injection. Estradiol serum level was determined in 8 day of cycle and when diameter of ovarian follicle was 10, 12, 16 mm and in the day of hCG injection. BMI and WHR was determined in the day of admission to hospital. RESULTS: The correlation between mean insulin level in the examined cycle and BMI and as well as the correlation between mean insulin level and WHR were determined. Mean level of serum glucose increased as the level of estradiol raised. No influence of growth level on insulin level was detected. PMID- 11883277 TI - [The place of laparoscopy in gynecological practice--useful technique for diagnostic and treatment of infertility and endometriosis]. AB - OBJECTIVE: The objective was to evaluate indications for laparoscopy performed in Department of Gynecological Surgery of Polish Mother's Memorial Research Institute. MATERIALS AND METHODS: We have analyzed hospital charts of 342 patients who had been treated with laparoscopy in 1991-1999. RESULTS: During last years a progressive frequency in performing laparoscopic procedures has been noted (4.7% of all surgical treatment in 1991-93, 6.4% in 1994-96, 16.3% in 1997 99, and 25.2% in 1999). An increased percentage of operative laparoscopic procedures has also been observed (11.9% in 1991-93 vs 59.8% in 1994-96 vs 86.9% in 1997-99). The major indication for performing laparoscopy was: infertility (145--42.4%), endometriosis (90--26.4%) and benign ovarian tumors. Most of laparoscopic operations were performed due to benign pathological changes found in adnexa (218--68.6%). Laparoscopy has been recently introduced in surgical treatment of uterine myoma (enucleation, laparoscopically assisted vaginal hysterectomy-LAVH) and diagnosis of ovarian carcinoma. Complications were found in 19 (5.5%) cases: 11 cases (3.2%) intraoperative and 8 (2.3%) cases postoperative. Bleeding from abdominal wall vessels was the most often intraoperative complications, whereas an infection was the most common postoperative one. Because of complications laparoscopy turned into laparotomy in four cases. CONCLUSION: Laparoscopy is a safe and useful technique for treatment some gynecological pathology (especially infertility and endometriosis) with a small number of complications. PMID- 11883278 TI - [The usefulness of hysteroscopy and hysterosalpingography in diagnosis of tubal infertility]. AB - OBJECTIVES: To estimate the usefulness of hysteroscopy (HSC) and hysterosalpingography (HSG) examination on a retrospective study. DESIGN: Canulisation and morphology of ovarian tube detected during HSG and reactivity of uterine tubal ostia observed in HSC were compared. MATERIAL AND METHODS: The results of HSG and HSC examination performed on 125 young infertile women whom the tubal uterine factor of infertility was suspected were analyzed 250 ovarian tubes, after previous selection were taken into consideration. The analysis had a retrospective character. RESULTS: The analyzed group of women were divided into 3 group which differed from each other in reactivate of uterine ostia of the ovarian tubes. The first group first with correct reactivity of tubal ostia was the largest. The authors observed canulisation, which was the best in the first group and morphologic changes in the internal structure of the ovarian tubes. The highest percentage of these changes were observed in group three with unreactive uterine ostia. CONCLUSIONS: The HSG and HSC examination are supplementary examinations. The use of both methods together increases their diagnostic value and gives a more correct estimation of tubal status. PMID- 11883279 TI - [Hysteroscopic correction of uterine malformations by small diameter hysteroscope with flexible needle]. AB - The series of 14 hysteroscopic surgery procedures in case of septum uteri were analyzed. All women enrolled into the study were infertile. The malformations were diagnosed by hysterosalpingography and intravaginal ultrasound. The laparoscopic examination of intrauterine serosal surface and operative hysteroscopy were performed in the same time. The 8 mm diameter hysteroscopy with flexible needle was used. The uterine cavity solution was distended by sorbitol and mannitol. The patients left the hospital on the second postoperative day. There was one complication in this study: intravasation of distended fluid. After one month from surgery the pelvis was controlled by HSG. In 12 cases the uterine cavity was restored to a normal shape, in 2 cases the residual uterine septum of less than one centimeter was determined. We concluded that correcting the septum uteri by the small diameter hysteroscope (8 mm) with flexible needle is an effective and safety method for the uterine septum treatment. PMID- 11883280 TI - [Serum and peritoneal fluid CA-125 concentration in women with endometriosis]. AB - OBJECTIVE: To evaluate the clinical utility of CA-125 measurements in peritoneal fluid (PF) as an indicator of endometriosis. DESIGN: CA-125 levels in paired serum and peritoneal fluid were assessed by IMx CA-125 Assay (ABBOTT). PATIENTS: 107 women with and without endometriosis, undergoing laparoscopy during the luteal phase of the menstrual cycle were selected for the study. RESULTS: CA-125 levels in peritoneal fluid were higher than those found in serum and were significantly elevated (P < 0.05) in comparison to the control group. In women with endometriosis stage I-II and stage III-IV. CA-125 levels in serum were increased only in stage III-IV. CONCLUSIONS: The sensitivity of the peritoneal fluid CA-125 test for endometriosis is higher than the respective serum test. The measurement of CA-125 levels in PF could be useful in the detection of the early stage endometriosis, which seems to be missed by the CA-125 serum test. PMID- 11883281 TI - [Menopausal medicine of the threshold of the third millennium]. AB - The paper reviews most important trends in hormonal replacement therapy which have appeared in recent years. The tailored approach to HRT, introduction of new drugs and modification of established treatment regimes are the most probable directions in which menopausal medicine would develop in the years to come. PMID- 11883282 TI - [Effects of oral administration of estrogen replacement therapy in surgical menopause]. AB - OBJECTIVE: The aim of this study was to analyze effects of oral administration of conjugated estrogens in women after surgical menopause. MATERIAL AND METHODS: A prospective study that included 41 women after hysterectomy with bilateral oophorectomy, was performed. All patients received conjugated estrogens (Premarin) 0.625 mg daily from second or third days after surgery. Plasma level of estradiol, FSH and several homeostatic parameters were evaluated before and after 2, 5 days and 6 weeks of hormone therapy. The symptoms of climacteric syndrome were estimated based on the scale of Kuppermann. RESULTS: A decrease in plasma estradiol levels in early postoperative days and an increase up to 100 pg/ml in estradiol concentration after 6 weeks of estrogen therapy were observed. Changes in platelet number, plasma concentration of coagulation factors were transient and there were no significant thrombophilic effects after 6 weeks of Premarin administration. CONCLUSIONS: The postoperative oral administration of conjugated estrogens did not cause thromboembolic complications among women after surgical menopause. Early beginning of estrogen replacement therapy after hysterectomy may prevent symptoms of climacteric syndrome occurrence. PMID- 11883283 TI - [Serum estradiol levels in early postoperative period after surgical castration- the influence of estrogen replacement therapy]. AB - In 40 premenopausal women who underwent TAH&BSO serum E2 levels were measured by means of RIA in 3 time points: before the operation, in the postoperative day 2 and in the postoperative day 3. In 20 of the subjects the ERT (transdermal estradiol 0.1 g/24 h) was introduced in the 2 postoperative day. In the postoperative day 2 the average serum E2 decreased by nearly 90% as compared to the preoperative values. Twenty four hours after the application of the transdermal E2 patch, the estradiol level returned to the values insignificantly different from the preoperative ones. PMID- 11883284 TI - [Changes of SHBG concentrations in postmenopausal women]. AB - Sex hormone-binding globulin (SHBG) is a serum (serous) glycoprotein binding estrogens and androgens with high affinity. Hepatic synthesis of SHBG and serous protein concentration are related to the following factors: age, sex steroids and thyroid hormones concentrations, obesity. Along with aging process, an increase of SHBG concentration is observed in both sexes. The following paper confirms a relation between age and SHBG concentration (positive correlation, p < 0.001) and a relation between body mass index (BMI) and SHBG. Concentration (r = -0.35, p < 0.039). In our studies it was found out that SHBG in postmenopausal women with recent hip fractures was significantly lower than in a group of women without fractures. The authors suggest that the determination of serum SHBG concentration might be assumed as a screening test for evaluation of the hip fracture risk in postmenopausal women. PMID- 11883285 TI - [Luteal and follicular phase inhibin A and B in regularly cycling women]. AB - OBJECTIVE: To investigate the level of inhibin A nad B in luteal and follicular phase in women of reproductive age. PATIENTS: Seventy women 39-52 years of age with regular menstrual cycle. INTERVENTIONS: Blood samples obtained on days 3-8 and on days 22-25 of menstrual cycle were assayed for FSH, estradiol, inhibin A, inhibin B. RESULTS: Luteal and follicular phase inhibin B was correlated inversely with age. Luteal phase inhibin A was correlated inversely with follicular phase FSH. CONCLUSION: Main form of inhibin in follicular phase of the cycle is inhibin B and in luteal phase inhibin A. Inhibin B can be potential marker of ovarian aging. PMID- 11883286 TI - [The atherogenic lipid profile parameters and fibrinogen serum concentration in climacteric women under various routes of hormone replacement therapy administration]. AB - OBJECTIVES: Influence of various routes of HTR on atherogenic lipid profile parameters and serum fibrinogen concentration was investigated. DESIGN: In 85 women in four groups receiving HRT transvaginally, transdermally, orally and intramuscularly, the total cholesterol, HDL-cholesterol, triglycerides, apoB, apoA, LpA and fibrinogen serum concentration was assessed before treatment and after six month. RESULTS: Most of tested parameters: total cholesterol, LpAI, apoB, LDL cholesterol, non HDL cholesterol, changed favorably (lowered concentration) in group using HRT orally and intramuscularly. CONCLUSION: Orally and intramuscularly route of administration HRT have better cardioprotective effects. PMID- 11883287 TI - [Men's acceptance of continuous hormone therapy]. AB - The aim of the study was the assessment of males' willingness to long-term administration of hormonal drugs in substitutive and contraceptive means. Eighty males in their third and fifth decade were inquired a questionnaire. The patients were divided according to age into two groups: A (20-30 yrs of age) 55 males, and B (41-50 yrs of age) 25 males. University education was a condition required to be enrolled into the study. The results was statistically analyzed with Chi 2 test. 65.5% of males in their third decade would not accept hormonal contraception. 40% of males declared their acceptance for hormonal substitutive therapy and they were also aware of the necessity of prophylactic examinations, 7.3% accepted the therapy but without regular follow-ups, 16.3% did not accept the therapy and 36.4% did not consider it as necessary. 56% of the respondents from group B would accept an administration of contraceptive pill. 44% would not accept hormonal substitutive therapy, 24% did not see the necessity of the therapy, 16% would accept the therapy and the necessity of regular prophylaxis, and the last 16% would accept the therapy, however without control examining. PMID- 11883288 TI - [Chromosome instability in women with genital organs carcinoma]. AB - OBJECTIVES: The aim of the current work was the assessment of the mutagen susceptibility of chromosomes of patients with carcinomas in comparison to healthy volunteers. It was interesting whether the bleomycin assay can be useful for searching for more susceptible to cancer disease individuals. MATERIALS AND METHODS: There were 4 groups of patients analysed: controls and three test groups (patients with uterine cervix carcinoma, endometrium carcinoma and ovarian carcinoma). In total 108 female patients were examined by use of bleomycin assay. Lymphocytes were cultured in vitro and treated with bleomycin. The b/c (breaks per cell) index was evaluated by use of light microscopy. RESULTS: Statistically significant increased test values were found in patients with uterine cervix carcinoma, endometrium carcinoma and ovarian carcinoma. CONCLUSIONS: The assessment of chromosome instability could be a useful prognostic test in the diagnosis of carcinoma of female genital organs. The bleomycin assay is useful for searching subpopulations with higher chromosome instability and more susceptible to cancer disease. PMID- 11883289 TI - [The risk factors of endometrial cancer]. AB - Authors presents the risk factors in endometrial cancer underlying such problems like hyperestrogenism, both external and internal caused by hormonally active ovarian masses, polycystic ovarian syndrome, adrenocortical hyperfunction and role of obesity in this pathological state. Other factors have been also described diabetes mellitus and hypertension, oral contraception, genetics factors, patient's obstetric history and other diseases where the increase of aromatization activity of androstendion to estron has been noted. PMID- 11883290 TI - [Panoramic hysteroscopy in prophylaxis of precancerous lesions and endometrial carcinoma]. AB - OBJECTIVES: The purpose of the study was to check the conformity of diagnosis based on macroscopic hysteroscopy image with the respective pathology report. MATERIALS AND METHODS: Study population consisted of 160 patients aged 45-86 years, with abnormal uterine bleeding or abnormal ultrasonographic image of endometrium. In all cases pelvic sonography and hysteroscopy were performed. RESULTS: Abnormal findings were detected hysteroscopically in 123 cases out of 160 hysteroscopies performed. Hysteroscopic diagnosis was confirmed by pathological examination in 100 cases (81.3%). The conformity of hysteroscopic image with the pathology report varied in different lesions, amounting to 90.9% in cases of submucosus myomas, 86.9% in endometrial polyps, 25.0% in endometrial hyperplasia and 71.42% in cases of endometrial carcinoma. CONCLUSIONS: 1. Hysteroscopic features are in high agreement with pathology report with reference to endometrial cancer, endometrial polyps and submucosus myomas. 2. Hysteroscopy does not seem to be a satisfactory tool for the differentiation between endometrial hyperplasia and endometrial carcinoma. PMID- 11883291 TI - [Diagnostic procedures of pathological bleeding in women population: comparison of hysteroscopy, ultrasonography and microscopic examination endometrium]. AB - The goal of this study was to asses the diagnostic value of hysteroscopy, cytology, ultrasonography and histopathology in various pathological states in endometrium. 250 patients with abnormal uterine bleeding were examined. The wide range of several diagnoses were achieved with 8 cases of cancer. Used methods were found to be complementary because 7 cases of cancer were recognised by histopathologic method, 6 by hysteroscopy and 5 by a cytological test (3 results were suspicious). To the risk group were qualified by ultrasonography all patients in postmenopausal age. PMID- 11883292 TI - [Histone H3 gene expression level as markers of hyperplasia simplex of endometrium]. AB - SUBJECT: The excessive proliferative activity of endometrial cells drives to hyperplasia, via states: simplex hyperplasia, complex and atypical hyperplasia, and finally preneoplastic state may arise. New molecular markers and their ability to differentiate physiological and pathological states are described. DESIGN: The aim of this study was to evaluate the usefulness of histone H3 gene expression level as a marker of hyperplasia of endometrium. MATERIAL AND METHODS: The level of H3 mRNA was estimated using RT-QPCR method in proliferative phase (n = 5) and endometrial hyperplasia simplex (n = 8). Differences in expression level of H3, were evaluated by Student's t-test (t-test for independent samples). RESULTS: In our experiments the average mRNA H3 expression was significantly higher(p < 0.05, p = 0.03) in case of hyperplasia simplex in comparison to H3 expression in proliferative state of endometrium. CONCLUSION: The results confirm advantage of estimation of H3 mRNA level as a marker of cell proliferation in endometrium. PMID- 11883293 TI - [The effect of progestins on endometrial cancer in morphometric evaluation]. AB - The morphometric estimation of the influence of the caproate 17-alfa hydroxyprogesterone on the endometrial cancer was the aim this work. The quantitative evaluation of the reaction of neoplasmatic and some stromal cells such as fibroblast, fibrocytes, lymphocytes, macrophages was carried out. Every patient was given a high doses of the medicine before the operation. The comparison of the histological image and than morphometric estimation was done after operation. Examination was performed in three separate groups depending on the grading of the tumor. The result was the decreased number of the neoplasmatic cells in every group independently on the grading. In the fibroblasts and fibrocytes groups the increased number of cells especially in the G1 and G2 group was observed. In the group of lymphocytes and macrophages the increased number of all cells was observed in every group. On the basic of our results we assume that the application of the high doses of progestins decreased number of neoplasmatic cells and stimulates the cells of the monocyte-macrophage system. PMID- 11883294 TI - [Serum cytokines in patients with ovarian cancer and benign ovarian cysts]. AB - The aim of our study was to evaluate serum interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) as tumor markers--study based on the data about tumor cytokines production and tumor-host interactions. METHODS: We investigated 48 women: 17 with ovarian cancer untreated before, 16 with benign ovarian cysts and 15 healthy controls. Venous blood for cytokines determinations were obtained before operations and during routine screening tests. Titers of cytokines were measured by means of ELISA technique. RESULTS: In the control group the upper limit of normal IL-6 titers (95th percentile) was 5.5 pg/ml; the mean IL-8 concentration was 9.6 +/- 15.1 pg/ml and the upper limit of normal was 37 pg/ml; serum TNF-alpha and IFN gamma were not detectable. In patients with benign ovarian cysts the levels of all investigated cytokines didn't differ significantly from healthy controls. Women with ovarian cancer had significantly higher serum IL-8 levels (mean: 290.5 +/- 351 pg/ml) than healthy controls or women with benign ovarian cysts; 88% of them had IL-8 titers above the normal. The IL-6 titers in ovarian cancer were also higher but didn't reach statistical significance, 53% of them had IL-8 titers above the normal. TNF-alpha and IFN-gamma levels in ovarian cancer were similar to patients with benign ovarian cysts. CONCLUSION: Serum IL-8 levels in patients with ovarian cancer were significantly higher when compared to healthy controls and benign ovarian cysts and in almost 90% of ovarian cancers the titers of IL-8 were increased. Additionally, 53% of women with ovarian cancer had elevated serum IL-6 levels. PMID- 11883295 TI - [Granulosa cell tumor--the assessment of some clinical and therapeutic parameters as prognostic factors]. AB - The results of the clinical and therapeutic factors in prognostic mean was presented. 48 cases of granulosa cell tumours treated from 1984 to 1994 in Oncology Centre in Warsaw were analysed. In investigated group 13 patients died, but only 8 because of relapse of the tumour. Among all analysed patients, 79% have reached 5 years free survival period. Tumour rupture, FIGO stage and incidence of irregular bleeding before recognition of the tumour had significant prognostic value. There were surprising that relative risk of relapse between patients stage I and II were similar (1.0 vs 1.01). The relative risk between I and III stage had strong prognostic difference. Additional operation after no radical surgery did not influence on better prognosis, but followed radiotherapy increase treatment results. PMID- 11883296 TI - [Identification of dendritic cells subsets in peritoneal fluid]. AB - Dendritic cells (DC) play a crucial role in the activation of naive T lymphocytes and in the generation of primary T cell responses. DC are present in lymphoid and non lymphoid tissues. Our previous data have shown, that DC are present in peritoneal fluid (PF). There is no other data in literature on this subject. The aim of our study was identification of DC in peritoneal fluid and peripheral blood patients with infertility (n = 5) and benign noninflammatory ovarian tumor (n = 16). Mononuclear cells from PF and peripheral blood were isolated on gradient density centrifugation (Lymphoprep, Nycomed-Norway). Isolated cells (106 of cells per tube) were incubated with mAbs. We collected 300.000 cells using a FACSCalibur flow cytometer and analysed with CellQuest Software. The following directly conjugated monoclonal antibodies were used: anti-BDCA-1(CD1c) FITC, anti BDCA-2 FITC (Miltenyi Biotec) and anti-CD19 CyChrome, anti-CD123 PE (Pharmingen, USA). The concentrations of peritoneal fluid leukocytes in infertility patients and women with ovarian tumor approximated 6.8 x 10(6) and 7.76 x 10(6), respectively. Using this method, myeloid DC comprised 8.08 +/- 2.69% mononuclear cells PF in infertile women and 11.23 +/- 6.59% mononuclear cells PF in patients with ovarian tumor. The average mean of lymphoid DC was 0.63 +/- 0.33% mononuclear cells PF in infertile women and 0.59 +/- 0.33% mononuclear cells PF in patients with ovarian tumor. The percentage of dendritic cells in peritoneal fluid was significantly higher than in peripheral blood in both studied groups (p < 0.05). PMID- 11883297 TI - [Women's opinion of the risk of breast cancer]. AB - The basic element of preparing primary prophylactic is the designation of factors of the risks. In this connexion it has been decided that we acquaint ourselves with the opinions of women. Regarding the factors of the risks of falling ill with breast cancer, to be found on them. The research has been carried out among 149 women in the period of procreation. In order to obtain the material required for the research we hare used the questionnaire of the poll of their proper ownership. The gathered material was subjected to a statistic and descriptive analysis. Most of surveyed (138, it. 92.6%) has estimated the degree of the risks of falling ill with breast cancer. The women associated this fact with the cases of falling in their families or the changes on their breast. When asked, what increases the risks oh falling ill with breast cancer in their it was connected with women's gynaecological and maternity post. CONCLUSIONS: 1. Over halt of the women (53.6%) has estimated the risks of falling ill with breast cancer giving 1 2 points (within the scale 0-5 points). 2. In the families of the surveyed there were cases of falling ill with malignant breast tumour (3%). 3. The vast majority (78.5%) undertakes the steps in order (wholesome falling in advantageous to their health and controlling their state of health) to protect themselves against tumourous disease. 4. The variables accepted while working did not differentiate the surveyed opinions. PMID- 11883298 TI - [Early diagnosis of gynaecologic and breast cancers]. AB - Gynaecologic malignant neoplasms are significant medical and social problem in Poland. The majority of Polish women suffer from breast, cervix, ovary and endometrial cancer. This paper shows common and new methods in early diagnosis of neoplasmic diseases. PMID- 11883299 TI - [Women in reproductive age in the face of prophylactic gynaecological examinations]. AB - In this study attitudes of young women towards prophylactic gynaecological examination were presented. 149 women were included into the questionnaire, all with secondary education in medical field. The most frequent reason of a first visit was the appearance of symptoms and sings, causing anxiety, and fear the second frequent reason was simply a check--visit. The majority of women (64.5%) confirmed the choice of one, regular doctor, to whom they used to report their various health problems. PMID- 11883300 TI - [Adenocarcinoma of the cervix--stage IB: results of treatment and prognostic factors]. AB - The study presents the results of treatment of 63 women with stage IB adenocarcinoma of the cervix. The treatment consist of radical Wertheim-Meigs hysterectomy with adjuvant brachytherapy or external beam irradiation. A statistical analysis with Kaplan-Meier and cox model was conducted to assess the influence of selected prognostic factors on survival. After 5-year long follow-up 73.4% of patients were alive without recurrence. The results of univariate analysis showed that metastatic pelvic nodes, cervical tumor size of more than 4 cm and the presence of microscopic parametrial infiltration had a statistically significant negative influence on survival. In a multivariate analysis tumor size of more than 4 cm and microscopic parametrial involvement had an independent negative impact on survival (relative risk of death--12.1 and 15.7). PMID- 11883301 TI - [Serum levels of proinflammatory cytokines in women with uterine myomas]. AB - The levels of proinflammatory cytokines, namely IL-1 alpha, IL-6 and TNF-alpha were determined in blood serum of 26 women with uterine myomas and in 20 healthy controls. A significantly elevated concentration of IL-1 alpha was found in patients with uterine myomas. Levels of IL-6 and TNF-alpha were insignificantly higher in women with myomas. The obtained results indicate the possible role of IL-1 alpha in the pathogenesis of uterine myomas--further studies are needed to elucidate the role of proinflammatory cytokines in the proliferative disorders of myometrium. PMID- 11883302 TI - [Detection of human papillomavirus genome in cervical squamous intraepithelial lesions]. AB - OBJECTIVES: The aim of this work was the assessment of the correlation between different types of HPV and abnormal stages of uterine cervix. MATERIALS AND METHODS: The smears were simultaneously collected for both cytooncologic examination and PCR identification of HPV viruses. Cytologic smears were classified according to the Bethesda system. RESULTS: A strong relationship between the presence of HPV types 16, 18 and 33 and the intensification of cytologic changes was confirmed. The more advanced abnormal changes in the uterine cervix, the more often presence of these HPV types. The presence of HPV types 6 and 11 were more often in low grade lesions. CONCLUSIONS: The conclusions from this examinations are: there is a proportional correlation between the grades of Squamous Intraepithelial Lesions, uterine cervix carcinoma and the presence of oncogenic types of human papillomavirus. PMID- 11883303 TI - [PCR and Digene Hybride Capture System I in identification of human papillomavirus]. AB - We have analyzed and compared two different systems used in identification of DNA HPV. 100 samples obtained from patients with I degree and II degree Papanicolaou smears and 200 from patients with cervical intraepithelial neoplasia were analyzed. The obtained data indicate higher sensitivity of the PCR method compared to Digene Hybride Capture System. PMID- 11883304 TI - [Primary vaginal and uterine melanoma--a case of long term survival after local excision and vaginal brachytherapy]. AB - This report presents the case of long term survival of primary vaginal melanoma treated by local excision and vaginal brachytherapy. A unique histopathological pattern of preinvasive vaginal melanoma is also described. A review of the literature revealed 22 long term survivals after treatment of malignant melanoma of the vagina, and only 4 surviving more than 10 years. In general the prognosis in women with these malignancy is poor regardless of type of surgery. Depth of infiltration seems to be the only important prognostic factor influencing the survival. PMID- 11883305 TI - [Analysis of urinary tract injury during gynecological surgery performed with preoperative ureter catheterization or intraoperative ureter control]. AB - DESIGN: To analyze urinary tract injuries during gynecological surgery performed with preoperative ureter catheterization or intraoperative ureter control. MATERIAL AND METHODS: Retrospective analysis of 1986 chosen gynecological operations performed between 1990-1998 in Dept. of Gynecological Surgery Polish Mother's Health Center Institute. RESULTS: The overall frequency of urinary injuries was 2.15%. It was less during gynecological surgery performed with ureter catheterization compared to intraoperative ureter control only (ureter: 0.30% vs. 0.55%, p = 0.22; urinary bladder: 0.40% vs. 0.90%, p < 0.05). CONCLUSION: 1/ urinary injury was twice as frequent during ureter control than during ureter catheterization, 2/ urinary injury was the most frequent complication during hysterectomy with adnexa, 3/ the results of our analysis should be treated as a vote "for" ureter catheterization before gynecological surgery. PMID- 11883306 TI - [Menstrual abnormalities in women with biliary cirrhosis treated with liver transplantation]. AB - A study presents two cases of menstrual abnormalities in women with primary biliary cirrhosis and chronic active hepatitis treated with liver transplantation. The gonadotropin, sex hormones levels and biochemical parameters were presented. The menstrual disorders we observed may be a consequence of high concentration of oestrogens in our patients. PMID- 11883307 TI - [The influence of antibiotic prophylaxis on the incidence of fever and wound infection after gynecological operations in patients with increased risk of infection]. AB - Antibiotic prophylaxis was applied to patients with high risk of infection undergoing: myomectomy m. Martin, procedures on the uterine adnexa, total and/or subtotal hysterectomy and laparoscopy. The influence of this prophylaxis on the incidence of fever and wound infection was observed. For vaginal hysterectomy antibiotic prophylaxis was not applied. The risk factors included: the history of at least 1 laparotomy, suffer from chronic adnexitis in the past, past thrombophlebitis, diabetes and obesity. Statistically significant decrease of wound infections among patients underwent total and/or subtotal hysterectomy was noted, the highest incidence of fever occurred after myomectomy m. Martin, there was no wound infection following laparoscopy, the rate of infections after vaginal hysterectomy was much lower than after abdominal hysterectomy. PMID- 11883308 TI - [Colposuspension during abdominal hysterectomy--own modification]. AB - OBJECTIVES: The aim of our study was to evaluate the clinical effectiveness of own modification of colposuspension using the musculofascial flap during abdominal hysterectomy. MATERIAL AND METHODS: The study group consisted of 57 patients operated on uterine and/or vaginal prolapse; 45 of them had also others pelvic gynecological disorders. Prolapsed or lowered vaginal walls were corrected by colposuspension or cervical suspension using musculofascial flap (made of the rectus abdominis muscle sheet and pyramidal muscle). The flap was sharply separated from the anterior wall of rectus abdominis muscle sheet going up from pyramidal muscle to umbilical region where it ended. Its end was sutured to vaginal vault and uterosacral ligaments. This gave a flattening of rectovaginal pouch and shortage of rectovaginal distance. RESULTS: The incidence of usual complaints as: hypogastric pain, uterine/vaginal prolapse feeling and vaginal dryness was significantly decreased after the operation. The quality of sexual functions was also improved. The incidence of urine incontinence and polyuria didn't change after the operation. CONCLUSION: The own modification of colposuspension during abdominal hysterectomy is an effective method of treatment in cases of uterine/vaginal prolapse with other pelvic gynecological disorders. PMID- 11883309 TI - [Hematometra in 76-year-old woman followed by DIC and the result of the carcinoma of cervix--a case report]. AB - This is a case report of a 76-year-old women with hematometra due to cervical cancer followed by DIC. To the authors' knowledge this is the first report of such case. PMID- 11883310 TI - [Menstrual cycle in women with Wilson disease before and after liver transplantation]. AB - Four cases of young women with Wilson's disease who underwent liver transplantation are presented. The menstrual disturbances associated with Wilson's disease are presented and discussed. PMID- 11883311 TI - [Expression of the survivin gene in the scar endometriosis and in normal human endometrium]. AB - The survivin gene encoding a novel inhibitor of apoptosis protein (IAP) is located on chromosome (17q25). It is expressed during development and in human cancer in vivo. Endometriosis in cicatrix after cesarean section represents the only localization of endometriotic implant which etiology is known. It is believed that during the surgical procedure when the uterine cavity is open there is a high risk of decidualized endometrium implantation. The aim of the study was to estimate the expression of the survivin mRNA in the scar endometriosis after cesarean section (n = 6) and perineum endometriosis (n = 2) and in normal human endometrium (n = 12, reference group) using reverse transcribed-polymerase chain reaction (RT-PCR). METHODS: Total cellular RNA was isolated from the tissues using TriReagent according to the manufacturer's protocol. Total cellular mRNA was transcribed into cDNA with reverse transcriptase (RT) then the survivin gene fragment (430 bp) was amplified with PCR using specific primers. The products of the PCR reaction were separated on 2% agarose gel. The survivin gene was expressed in 4 of 6 cases of scar endometriosis after cesarean section and in 2/2 cases of perineum endometriosis. In all cases of normal endometrium we found the expression of survivin with a peak in the late proliferative phase. The antiapoptosis function of this gene product may play an important role in the endometriotic implant growth promotion. PMID- 11883312 TI - [Quantitative (RT-QPCR) analysis of telomerase subunit in normal endometrial tissues]. AB - Telomerase is a specialized ribonucleoprotein polymerase that elongates telomeres. To clarify the molecular mechanisms through which telomerase is activated in normal endometrium, we examined the expression profiles of each telomerase subunit in this tissues. A total of 22 normal human endometrium in various menstrual phases were examined for the expression of each telomerase subunit (hTERT, hTR, TP-1) using RT-QPCR. The hTERT as well as TP-1 subunit had higher expression levels in proliferative than secretory phase endometrium. Expression levels of hTR did not change during menstrual cycle. Our data showed the strong correlation between the level of expression of telomerase in human endometrium and phase of menstrual cycle. PMID- 11883313 TI - [Assessment of the PCNA and P53 proteins expression in the proliferative, hyperplastic and neoplastic human endometrium]. AB - In the current study, PCNA and p53 proteins were immunohistochemically studied in the proliferative (n = 5), hyperplastic (n = 4) and neoplastic (n = 20) human endometrium. PCNA immunostaining was noted in 2 out of 5 (40%) proliferative, 4 out of 4 (100%) hyperplastic, and in 18 out of 20 (90%) neoplastic slides. Concomitant PCNA and p53 expression was reported in 12 out of 20 (60%) malignant tumors. All non-endometrioid neoplasms were PCNA-positive, suggested this proliferative marker is commonly expressed in the unfavorable histological types of endometrial cancer. PMID- 11883314 TI - [Growth hormone, insulin, IGF-1 and estradiol levels during maturation of internal sex organs before and at menarche]. AB - INTRODUCTION: The growth hormone (GH) is the main activator of growth, exerting its biological effects through IGF-1 and IGF-2 growth factors. Secretion of GH is partly controlled by steroid sex hormones. MATERIAL AND METHODS: Body mass, height, BMI, and body composition were recorded every three months in 45 healthy girls 8 years of age. Tertiary sex features according to Tanner were determined. Transabdominal ultrasound of the uterus and ovaries was done. GH, insulin, and IGF-1 levels in serum were measured radioimmunologically with commercial test kits. E2 levels were determined with an immunoenzymatic method. RESULTS: GH levels increased until stage M3 of breast maturation and gradually decreased during stage M4 (12 girls with menarche). IGF-1 and insulin levels continued to increase until stage M4. Uterine and ovarian dimensions correlated with levels of hormones studied. CONCLUSION: High levels of GH and IGF-1 during the early and central stages of puberty seem to affect the maturation of internal sex organs. PMID- 11883315 TI - [Differences in human and rat FSH receptors promote activity as a result of the transcriptional factors: E2F1, E2F4 and E2F5 overexpression]. AB - FSH-R expression in granulosa cells varies during the course of ovarian ontogenesis, as well as, during each menstrual cycle. Expression of follicle stimulating hormone receptors (FSH-R) on Sertoli cells of the testis and ovarian granulosa cells depend on many paracrine and autocrine factors. The modulation of FSH-R synthesis is accomplished via a number of mechanisms including regulation of the promoter activity. Little is known about factors involved in control of FSH-R transcription in different species. The aim of the present study was to investigate differences in the regulation of human and rat FSH-R promoter activity by E2F transcriptional factors. MATERIAL AND METHODS: The 5'-flanking regions of human and rat FSH receptor gene subcloned in the pGL3 plasmid were transiently transfected into cultured CHO cells and rat granulosa cells. Rat granulosa cells were obtained by puncturing ovaries from DES primed immature Sprague-Dawley rats. Promoter activity was determined by measuring firefly luciferase luminescence of the cell lysate. Transfection efficiency was normalized by the renilla luciferase activity generated by co-transfected pRL-CMV vector. In order to determine the influence of E2F1, E2F4 and E2F5, on FSH-R promoter activity, cells were transfected either with promoter construct alone or with its mixture with selected expression vector. RESULTS: Rat FSH-R promoter construct (-1033/+6 bp) and human FSH-R promoter construct (-1485/-1 bp) were both active in transfected cells. Overexpression of E2F1 protein decreased both, human and rat wild type FSH-R promoter activity. Overexpression of E2F4 did not affect neither rat nor human FSH-R gene transcription. Expression vector for E2F5 increased both, human and rat, FSH-R promoter activity. Folds of increase were markedly higher in case of rat FSH-R construct transfection, comparing to human FSH-R promoter. CONCLUSIONS: Results suggest, that the E2F1 and E2F5 factors might play an opposite role in the regulation of FSH-R promoter activity. More pronounced stimulatory effect of E2F5 on the rat FSH-R can be explained by the presence of E2F site in the promoter. Since there is no E2F sensitive element in the human FSH-R promoter sequence, E2F1 and E2F5 can also indirectly influence FSH-R promoter activity. PMID- 11883316 TI - [The influence of hypoxia on the proliferation of endothelial cells originating from human umbilical vein (HUVEC)--an in vitro study]. AB - In this study we took for culture the early-passage human umbilical vein endothelial cells during 12, 24 and 36 hours of hypoxia. To assess the relative contributions of hypoxia, we compared the rates of BrDU incorporation into DNA of proliferating endothelial cells under normoxix and hypoxic conditions. Significant differences in proliferation rates were found only between 24 hours hypoxic group and the control group. Our results support the hypothesis that increased proliferation of human umbilical vein endothelial cells may reflect an adaptation of fetoplacental circulation to hypoxic conditions. PMID- 11883317 TI - [Dental care during pregnancy]. AB - In the paper authors discussed the importance of dental care during pregnancy, prophylactic methods, dental treatment procedures and the influence of oral health status on pregnancy and fetus. PMID- 11883318 TI - [Sialic acid of glycoconjugates in amniotic fluid]. AB - OBJECTIVES: Sialic acid is a negatively charged monosaccharide attached to non reducing end of N- and O-linked carbohydrate chains of glycoconjugates. The claimed biological functions of sialic acid include its participation in cell to cell recognition and interaction as well as affecting the function of receptors by providing binding sites for ligand. Increased sialic acid concentration have been observed in several diseases e.g. malignancies, diabetes, inflammatory disorders, rheumatoid arthritis and alcoholism. DESIGN: The aim of the present work was to determine if the amount of sialic acid attached to glycoconjugates of amniotic fluid changes during pregnancy. MATERIALS AND METHODS: The sialic acid content in 47 samples of amniotic fluid derived from pregnant women with gestational age between 13 and 42 was studied by sialic acid specific lectins immunosorbent assay. The patient samples were divided into seven groups. RESULTS: Time dependent changes in the degree of sialylation of glycoconjugates in amniotic fluid during pregnancy, particularly in advanced pregnancy were observed. Moreover, the highest sialic acid content on glycoconjugates in pregnancies complicated by premature rupture of membranes and is prolonged pregnancy were also detected. CONCLUSIONS: Sialic acid content determination in amniotic fluid could be a potentially useful marker of inflammation process of amniochorion during pregnancy. PMID- 11883320 TI - [Seventh caesarean section with the same woman--case report]. AB - Seventh caesarean section in the same pregnant woman was described. Pregnancy was terminated in 36 week of gestation cause of preterm uterine activity, after getting pulmonary maturation and estimating ultrasound fetal weight over three kilograms. The caesarean section and postoperative period were uncomplicated. There is little literature about multiple repeat caesarean sections. The authors discuss changes in outcomes and complications of multiple caesarean sections through last three decades. PMID- 11883319 TI - [Nocturnal fall of blood pressure in the first half of pregnancy]. AB - OBJECTIVES: Arterial blood pressure (BP) shows circadian variation. The most important feature of the circadian variation of BP is it fall during sleep. Nocturnal fall of BP achieves at least 10% ("dip"). Very scanty literature is available on the diurnal BP pattern in healthy gravidae with uncomplicated pregnancy. DESIGN: The aim of the study was to evaluate of the day-time/night time mean arterial blood pressure (MAP) fall in pregnant woman in the first half of pregnancy in comparison with healthy non-pregnant women. MATERIALS AND METHODS: Fifty seven healthy woman were included into the study, and divided into two groups: 23 pregnant woman in 7-20 weeks of first gestation and 34 non pregnant woman. All the subjects were included into the 24-hours non-invasive automated blood pressure monitoring (ABPM). RESULTS AND CONCLUSIONS: Nocturnal fall of BP appeared to be the most pronounced element of the circadian BP pattern in the studied women. Women in the first half of pregnancy exhibit reduction of BP in the night-time similar to that observed in the non-pregnant subjects. In average, 2/3 of pregnant women below 20 weeks of gestation belong to "dippers" in the MAP range. Women with higher mean diurnal BP value have less pronounced nocturnal BP "dip". PMID- 11883321 TI - [Sonographic evaluation of the uterus on 3rd day after normal, vaginal delivery in women with benign fever]. AB - OBJECTIVE: Sonographic evaluation of the uterus on 3rd day after normal, vaginal delivery in women with benign fever. MATERIALS AND METHODS: 112 primiparous patients after vaginal delivery on term, following uncomplicated pregnancy. Group I (n = 78, 69.6%) with body temperature below 37.5 degrees C, group II (n = 34, 30.4%) with temperature higher than 37.5 degrees C. In all patients following sonographic features were evaluated: uterus volume, uterine cavity volume and uterine cavity content. RESULTS: No significant differences were observed. CONCLUSIONS: We suggest, that uterus volume, uterine cavity volume and uterine cavity content is not often correlated with benign fever following vaginal delivery. PMID- 11883322 TI - [Nuchal cord and nuchal translucency measurements between 10th and 14th weeks of gestation]. AB - Our objective was to determine the incidence of nuchal cord and it's possible influence on the nuchal translucency (NT) measurements between 10th and 14th week of gestation. One hundred fourteen singleton pregnancies were studied with transvaginal sonography. Nuchal translucency was measured according to the criteria published by Snijders et al. from the Fetal Medicine Foundation. Nuchal cord was detected with color Doppler flow mapping in 8 of 114 fetuses (7.01%). Mean cord thickness in these fetuses was 1.1 mm (range 0.5-1.4 mm). Mean nuchal translucency with and without umbilical cord were 1.67 mm (SD = 0.67; range: 0.5 3.6) and 1.59 mm (SD = 0.54; range: 0.5-3.0), respectively. The differences were statistically significant (p = 0.0146, Student's t-test for paired observations). We conclude that although relatively infrequent, nuchal cord might have influence on nuchal translucency measurements at the end of the Ist trimester. PMID- 11883323 TI - [The effect of negative emotion on eyewitness functional field of view]. AB - This study examined whether the functional field of view was shrunk by negative emotion. The functional field of view was determined in terms of the number detection task, in which a number presented for a brief exposure peripherally in one of the four video screen corners. Participants watched either negative emotional or neutral event on the screen. Emotion was evaluated by means of self ratings of mood adjective checklist, and the results showed that tensional arousal increased in participants who watched the negative emotional event. After watching the video participants were asked to report whether they noticed the number. Participants who watched the emotional event were able to detect fewer numbers than participants who watched the neutral event. This result suggests that the functional field of view shrank because of negative emotion. Peripheral memory decrement reported in previous studies might be due to the shrinkage of the functional field of view. PMID- 11883324 TI - [Effects of environmental change and others' behavior on cooperative behavior and solution preference in social dilemma]. AB - This study examined how environmental change and others' behavior affected cooperative behavior and solution preference of the person in social dilemma situation. Participants in two experiments played an "environment game," in which gradual pollution in environment and reduction in profit rate were simulated. Information on behavior of other players was manipulated: in "free rider" condition, one person was an extreme free rider, and the others were cooperative; in "loafing" condition, everyone loafed. In both experiments, "Bad Apple Effect" was not observed clearly, and cooperative behavior increased as environmental pollution worsened. In Experiment 2, there was no main effect of others' behavior on solution preference. However, significant correlations were found among solution preference, motivation to control others' behavior, and perceived seriousness of the situation, only when an extreme free rider was among them. PMID- 11883325 TI - [The sources of self-knowledge and the cultural view of self]. AB - A number of 1,604 respondents, divided into six age-categories, answered a questionnaire to examine the sources of self-knowledge as defined and measured by Schoeneman's method, and the cultural view of self measured by Takata's scale. The results revealed that adolescent categories displayed the strongest self critical and interdependent tendencies, but referred to social sources (e.g., social feedback and social comparison) for self-knowledge least. On the contrary, adult categories were the most likely to refer to social sources in spite of their independent and self-enhancement tendencies. The results suggest that, positive self-images depend strongly on reference to social sources in Japanese culture, and that the independent construal of self, which is a dominant characteristic of Japanese people, is highly active in the adolescence period. PMID- 11883326 TI - [Three-dimensional curve tracing]. AB - The time required in judging if two probes are on the same curve increased monotonically with the separation of the probes along that curve. This process is called "curve tracing" (Jolicoeur, Ullman & Mackay, 1986). In this study we examined whether curve tracing would occur on a three-dimensionally presented curve with depth variation. By comparing the performance on depth varying and no depth varying stimuli, we examined the properties of three-dimensional curve tracing. The mean RT on three-dimensional stimuli increased monotonically as the distance between two probes increased, which indicates that curve tracing also occurs on three-dimensional stimuli. The mean RT on three-dimensional stimuli was longer than that on two-dimensional stimuli. Our results suggest that three dimensional structure of the stimuli caused additional costs on curve tracing. PMID- 11883327 TI - [A confirmatory positioning analysis: a multivariate analysis for finding change of image and evaluation of cognitive map]. AB - A confirmatory approach to the positioning method (Toyoda, 2001) that can be used for analyzing three-mode multivariate data measured by the semantic differential (SD) method is considered. Two examples of the confirmatory method are given to demonstrate the applicability of the model. The first is a method to test the difference of image, between an experimental group and a control group. The second is a method to evaluate cognitive maps that are drawn subjectively. The conventional standard methods are not capable of solving these problems. PMID- 11883328 TI - [Characteristics of information decay in short-term visual memory and a diffusion model]. AB - The effect of exposure duration of test stimulus on the decay rate in short-term visual memory (STVM) was investigated in a same-different task. In the model, it is assumed that memory noise, the variance of each convex value represented in STVM, increases with time, which causes the decay in STVM. In recognition experiments, the results showed that the decay rate is lower for longer exposure duration regardless of pattern complexity. Furthermore, the decay lasted gradually for 16 s with exposure duration of 1,200 ms, which suggests that prolonged exposure largely acts to reduce the decay rate but does not prevent the decay itself. These experimental data were well predicted by the model, which clearly indicates that memory noise increases as a linear function of retention interval and that the rate of increase of memory noise is inversely proportional to exposure duration. These results are interpreted by an extension of the integration model of Signal Detection Theory: The number of mental scanning, N, monotonically increases with an increase in exposure duration, s, and increase of memory noise is inversely proportional to N, so memory sensitivity d' is predicted to be multiplied by square route of s. PMID- 11883329 TI - [Trust and cooperation: an experimental study of PD with choice of dependence]. AB - This study examined the relationship between cooperation and trust in interpersonal trust formation. Previous studies of trust and cooperation using prisoner's dilemma (PD) games failed, methodologically as well as conceptually, to distinguish the former from the latter. In response to the criticism on the use of iterated PD games, and for the purpose of investigating dynamic relationship between trust and cooperation, an improvement in research methodology was recently proposed: namely, PD with choice of dependence (PD/D; Kakiuchi & Yamagishi, 1997; Yamagishi & Kakiuchi, 2000). We conducted an experiment to compare formation of trust relations in PD and PD/D. Result indicated a higher level of cooperation in PD/D than in PD. Further analysis of strategies used to build trustful relationship, where two partners trusted each other and reciprocated the other's trusting behavior, revealed that participants in PD/D adopted "cautious and unconditional cooperation strategy" rather than TFT strategy. PMID- 11883330 TI - [Cross-task analysis of age differences in constructive activity]. AB - The present study investigated age differences in constructive activity, using three different tasks: the Rey-Osterrieth Complex Figure, Block Design, and Benton's 3D Construction tests. Participants were 116 healthy adults, ages ranging from 25 to 74 years. Results of the ROCF test failed to show any age difference in test scores among the participants up to early 60's, task completion time among those up to early 50's, and strategy among all participants. However, results of the Block Design test showed age differences in test score, task completion time, and strategy among all participants. Results of the 3D Construction test failed to show any age difference in test score among the participants up to early 60's, task completion time among those up to early 50's, but showed age differences in strategy among all participants. These findings suggest that it is necessary to take into account task-specific age differences in constructive skills, when evaluating constructive disabilities. PMID- 11883331 TI - [Types of comprehension processes and the preliminary knowledge-use effect: analogical transfer of learning from text]. AB - This study examined what types of comprehension strategies promote the spontaneous transfer in a later problem solving task. In Experiment 1, 40 undergraduates read and comprehended an abstract text (cooperatively with a partner or individually) with a think-aloud method, and solved a target problem later. The comprehension processes for the text were found to be classified into three categories; GID (generate examples, integrate them, and develop them into other examples) type, GI (generate examples and integrate them, but not develop them into other examples) type, and GN (generate examples but not integrate them) type. The learners of GID type performed better than the others in the transfer task. In Experiment 2, 66 undergraduates read one of three types of texts composed in accordance with the comprehension processes found in Experiment 1, and solved the target problem. None of the types of texts promoted spontaneous transfer in the target problem. These results suggest that learning with GID strategy leads to spontaneous transfer only if learners spontaneously produce elaborations. PMID- 11883332 TI - [Performance on Matching Familiar Figures Test, classroom behaviors, and school achievements of elementary school children in Japan]. AB - This study investigated ecological validity of Cairns and Cammock's (1978) MFF 20, a revised version of Matching Familiar Figures Test, as an instrument to assess reflection-impulsivity. Data of 162 first-grade and 177 fourth-grade Japanese school children were analyzed, and partial correlations were computed among the variables: two MFF-20 subscores (Impulsivity and Efficiency), teacher ratings of classroom behavior and school achievement. The last two were measured twice, over a two-year period, as criterion variables. The effect of intelligence, assessed with Kyoken Group Intelligence Test, was partialed out. Analysis of partial correlations showed that the Impulsivity score of the first graders had stronger correlations than their Efficiency score with the criterion variables for their first and second grades. On the other hand, it was the Efficiency score of the fourth graders that became more dominant, in terms of correlations with the criterion variables, which were assessed in the fourth and fifth graders. PMID- 11883333 TI - [Gender stereotypes arising in a state of gender awareness]. AB - This study examined the structure of gender stereotypes which might arise in the state of gender awareness that was triggered by social situations where people perceived their gender differences strongly. Out of 1,500 residents in Tokyo aged between 20-60, 342 females and 313 males were randomly chosen and answered the questions about gender consciousness in the state of gender awareness. A factor analysis revealed that "maternity" and "trustworthiness" were the dominant dimensions of gender stereotypes in the state of gender awareness, and that trustworthiness particularly formed the basis of gender stereotypes. Generation differences in gender stereotypes were also revealed between women in their 40 s and 50 s, and between men in their 30 s and 40 s. Generally, power for men and nurture for women were more likely to be perceived in a state of gender awareness. PMID- 11883334 TI - A study of the effect of tamoxifen on serum lipoprotein profiles in premenopausal and postmenopausal women with breast carcinoma and associated risk of cardiovascular disease. AB - To examine the effect of tamoxifen on serum lipoprotein profiles of premenopausal and postmenopausal patients of breast carcinoma (without and with cardiovascular disease) we performed a short term evaluation of serum lipoprotein profiles of 38 pre and 42 post menopausal subjects of breast carcinoma (without and with cardiovascular disease) at baseline and after 3 and 6 months of tamoxifen therapy. The serum lipoprotein profiles of premenopausal patients of breast carcinoma, both without and with cardiovascular disease, showed no significant variation, after 3 and 6 months of tamoxifen treatment than the corresponding baseline values of premenopausal subjects. However, in postmenopausal subjects of breast carcinoma (both without and with cardiovascular disease), serum TC, Apo-B, and Lp (a) were significantly decreased and serum TG, HDL and Apo A1 were significantly elevated, after 3 and 6 months of tamoxifen treatment, than the corresponding baseline values of postmenopausal subjects. Also, the comparison of the results of the present study for pre and postmenopausal patients of breast cancer revealed that the administration of tamoxifen, as an adjuvant therapy for breast cancer, is estrogenic and beneficial for postmenopausal patients of breast carcinoma (both without and with the risk of cardiovascular disease) as the drug minimises the risk of cardivascular disease by bringing significant improvement in serum lipoprotein profiles of the patients. But the drug fails to bring any significant beneficial effect on serum lipoproteins of hyperiipoproteinemic patients of breast cancer. PMID- 11883335 TI - Comparison of ELISA for antibody detection and biopsy urease test against H. pylori in cases of gastoduodenal disorders. AB - Out of 156 cases of various gastro duodenal disorders studied H. pylori was diagnosed in 119 (76.28%) as indicated by Biopsy urease test and IgG ELISA. Biopsy urease test detected higher number of cases 119 when compared to IgG ELISA 107 cases. ELISA being a non invasive technique can be used successfully for the diagnosis of H. pylori infections. PMID- 11883336 TI - Incidence of anaerobes in throat infections and their sensitivity pattern in Ludhiana. AB - Of 175 throat swabs processed, anaerobes were isolated from 16 (9.14%) patients. Isolation of anaerobes from healthy controls was 2 out of 25 (8%). Peptostreptococci and Bacteroides species were the commonest isolates followed by Peptococci and Propioni-bacterium. All of these isolates were sensitive to Metronidazole. Clindamycin, Erythromycin and Tetracycline also showed good response. PMID- 11883337 TI - Antibacterial activity of black tea (Camelia sinensis) extract against Salmonella serotypes causing enteric fever. AB - Alcoholic extract of black tea (Camelia sinensis) was assayed for its antibacterial activity against Salmonella serotypes causing enteric fever viz., Salmonella typhi and Salmonella paratyphi A. While all strains of S. paratyphi A tested were found sensitive, only 42.19% of S. typhi strains were inhibited by this extract. Further minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of black tea extract against S. paratyphi A was less compared with that against S. typhi. PMID- 11883338 TI - Drug resistance among tubercle bacilli from pulmonary tuberculosis cases in central India. AB - An estimate of drug resistance is extremely important in the epidemiology and control of tuberculosis. Data on drug resistance among mycobacterial isolates from sputum samples analysed at Microbiology dept. of Choithram Hospital and Research Centre, Indore, M.P. is presented here. Drug sensitivity testing was carried out on 1426 Mycobacterial isolates by the method of proportion using critical concentration in Lowenstein Jensen medium. Resistance for Isoniazid, streptomycin, and pyrazinamide was found to be high (54.2%, 41.5% and 50% respectively) and was followed by resistance to rifampin (25%) and ethambutol (22%). Resistance for kanamycin, p-aminosalicylic acid, thiacetazone and ciprofloxacin was much lower (18%, 13%, 6.5% and 3.6% respectively). Only 12% of the isolates were sensitive to all the anti-TB drugs while resistance to two, three, and four or more drugs was in the range of 20-25%. Pattern wise, simultaneous resistance to INF and Rifampin with or without resistance to other drugs was observed in 8.1% while resistance for Isoniazid + pyrazinamide and Isoniazid + streptomycin was 11.9 and 11.5% respectively. Resistance for Isoniazid + ethambutol was the lowest (5.1%). Growing multiple drug resistance among tubercle bacilli warrant urgent attention in tuberculosis control programme. PMID- 11883339 TI - Contaminated antiseptics--an unnecessary hospital hazard. AB - A total of 284 antiseptic solutions were studied to check for their sterility. The overall antiseptic contamination rate was 15.14%. 14.85% of freshly prepared antiseptics were contaminated. Here, the problem could be attributed to inadequate precautions while preparing the antiseptics. 15.3% of the in-use antiseptics were contaminated. This could be due to improper handling. Non fermenters (45.45%), Pseudomonas aeruginosa (30.30%) and Klebsiella spp. (22.72%) were the commonest organisms recovered from the antiseptics. In 44.44% of patients, the isolates obtained from the catheterised urine in the same wards matched with the isolates from antiseptics of that ward. Antiseptic solutions have to be regularly monitored. If they are found to be contaminated, they should be discarded immediately and replaced by fresh sterile antiseptics otherwise instead of preventing infection, antiseptics will become a source of hospital acquired infection. PMID- 11883340 TI - Percutaneous minimal osteosynthesis of fractures of the proximal humerus in elderly patients. AB - INTRODUCTION: The aim of this study was to evaluate the subjective and objective outcome of the percutaneous minimal osteosynthesis in elderly patients. Untreated, unstable and dislocated proximal humeral fractures show poor functional and subjective results. Reduction and fixation of the fragments is essential to achieve a good clinical outcome. Especially noted in elderly patients, the osteosynthesis is concurrent with the implantation of a prosthesis. PATIENTS AND METHODS: We examined 31 patients with a mean age of 72 years (51-87) after an average follow-up period of 15.8 months (9-31). To assess the functional and subjective results, we used the Constant Score as well as an adapted version of the Oxford Shoulder Score. Radiographs in two planes displayed the anatomical situation, the healing of the fracture, and the evidence or absence of avascular necrosis. RESULTS: There were no local complications such as hematomas or wound infections. The results of the Constant Score showed an average of 63.5 Pts. (18 82 Points) and adapted to age and gender by 90.2% (28.1-118.8%). Compared with the uninjured arm with an average of 77.1 Points (20-89 Points) and 109.5% (29 129%) respectively, we achieved an average functionality of 82.6%, operated versus non-operated side. All but one fracture, that showed signs of avascular necrosis with disintegration of the humeral head, were fully consolidated. The subjective results were very good in 18 patients, good in 7, satisfying in 3 and poor in 3 patients. CONCLUSION: Our data indicate that the percutaneous minimal osteosynthesis is a valuable method for the fixation of proximal fractures of the humerus in elderly patients. The technique has a very low rate of complications and the time of convalescence is short. If closed reduction fails or a stable percutaneous fixation of the fracture by K-wires is not possible, the change to open reduction and internal fixation or the implantation of a prosthesis is required. PMID- 11883341 TI - [Carpal tunnel syndrome after trauma]. AB - The carpal tunnel syndrome is a frequent illness with several etiological factors. Its appearance after trauma is rare. In a retrospective study its incidence and the trauma pattern were analyzed. From 1.1.95 to 31.12.99 144 median nerve decompression procedures for carpal tunnel syndrome were performed in 114 patients. Twelve patients (10.5%) had suffered a trauma in the recent or more distant past. There were six distal radius fractures, three metacarpal fractures, one finger fracture, one humeral shaft fracture and one distal avulsion of the biceps tendon. In eight patients the symptoms appeared 1-3 months after trauma, in four patients there was an interval of several years. In all twelve patients electroneurography revealed pathological parameters on the symptomatic side, but in ten patients the contralateral side was also affected although there were no symptoms. According to the criteria given by Assmus and Frobenius [1], five patients showed an obvious and three patients a possible posttraumatic carpal tunnel syndrome. In four patients a distinct relation to the trauma could not be proven. CONCLUSION: The carpal tunnel syndrome after trauma is rare. Given the fact that the contralateral side in these patients was affected as well, a predisposition--due to a narrow carpal tunnel--is very likely. Its manifestation might be triggered by a pressure increase in the carpal tunnel as result of the trauma. PMID- 11883342 TI - Feasibility and potential of MR-Colonography for evaluating colorectal cancer. AB - BACKGROUND: MR-Colonography (MRC) based on MR-imaging is a relatively new diagnostic modality for diagnosing colorectal polyps. The aim of this experimental study was to evaluate its performance in detecting and staging colorectal cancer. METHOD: 23 patients with proven colorectal cancer underwent MR Colonography one day prior to operation after standard bowel preparation. The colon was filled with a diluted gadolinium enema. Coronal sections were acquired in prone and supine. Virtual colonoscopy (VC) was processed from the acquired 3D data sets and MRC was interactively analysed together with VC. The findings were correlated with colonoscopic and pathology results. RESULTS: A complete MRC was achieved in 21 patients (92%), two patients (8%) could not be conclusively evaluated due to insufficient bowel preparation or technical problems. No complications were observed. Compared to colonoscopy all 23 carcinomas were detected. No lesion > or = 8 mm was missed. In one patient a synchronous carcinoma was newly diagnosed. This lesion was missed by colonoscopy since the distal tumour was endoscopically unpassable. In total eight (33%) colonoscopies were incomplete. CONCLUSION: MRC offers a new and promising diagnostic tool for colorectal cancer. It is particularly valuable when colonoscopy is incomplete. It improves preoperative planning and it holds the potential as an all-in-one investigation including local and liver staging in combination with conventional MRI. PMID- 11883343 TI - [Should benign tumors of the liver be operated?]. AB - OBJECTIVES: To define a strategy for benign liver tumours treatment and to report our experience. DESIGN: A retrospective study. PATIENTS: Sixty-eight patients operated for benign liver tumours among 424 operated of liver resection. RESULTS: Benign liver tumours were discovered during radiologic exploration for upper abdominal pain in 69%, incidentally during radiologic examination in 23% or at laparotomy in 8% of patients. Preoperative assessment of solid benign liver tumours was based on 4 radiologic examinations in average whereas cystic lesions needed only 2. The surgical treatment ranged from a simple tumorectomy to liver transplantation depending on the type, size and location of the tumour. There were no postoperative deaths and minor complications occurred in 7 patients (10%). Twenty hemangiomas, 20 hepatic cysts, 22 focal nodular hyperplasias (FNH), 4 hepatic adenomas (HA) and 2 cystadenomas were found on histology. The final diagnosis differed from the preoperative one in 36 patients (53%). CONCLUSIONS: Differential diagnosis for cystic lesions are the hepatic cyst, the hydatid cyst and the cystadenoma. Hydatid cysts and cystadenomas are resected whereas the asymptomatic hepatic cyst is left untreated. Hemangiomas must be resected only when symptomatic or pedunculated. The differentiation between FNH, HA and hepatocellular carcinoma is difficult and final diagnosis is most often achieved only by surgery. PMID- 11883344 TI - [Stress fractures of the cuboid bone: an easy to treat rarity]. AB - Stress fractures of the cuboid bone are very rare. As in our present case these fractures are seen mostly in ambitious sportsmen and women. The symptoms described are nonspecific. The case history and a clinical examination, in combination with a conventional x-ray picture, should provide evidence of such a suspected fracture. Bone scintigraphy, a CT scan or magnetic resonance imaging may be necessary to confirm the diagnosis. Treatment is based mainly on immobilisation by means of a plaster cast of the lower leg or, if necessary, with ready-made splints. In the present case, healing was obtained by conservative measures after a few weeks. Also important is appropriate adaptation of the patient's further sporting activity, if necessary supplemented by foot-orthopedic measures such as arch supports or correct footwear. Especially in sports involving running, the importance of optimal footwear for the treatment or prevention of problems of overstrain is well known. PMID- 11883345 TI - [Prospective assessment of the learning curve and safety of stapler hemorrhoidectomy]. AB - INTRODUCTION: Stapled haemorrhoidectomy (SH) is a recently introduced procedure for the surgical excision of haemorrhoids. Actually, there is only limited information concerning the impact of the learning curve, complication rates and long-term results. Therefore, a prospective single-center study was performed with special regard to the learning curve and clinical safety of SH. METHODS: The data of 61 SH performed between March 1999 and May 2001 were analyzed. Operating times, complication rates and outcome results were prospectively recorded and then correlated to the surgical experience of the operating team. Postoperative pain was measured using the visual analogue scale (VAS). Sphincter lesions represented by the patient's incontinence and muscle defects were analyzed by using Williams incontinence score and histological examination of the resected specimen. Clinical follow-up studies were performed three and twelve weeks postoperatively. RESULTS: There were 18 patients with grade II haemorrhoids, 38 patients with grade III haemorrhoids, and five patients with grade IV haemorrhoids. Both, operating times and complication rates decreased with more surgical experience. The mean pain score during the first four postoperative days was 1.9 (range 0-8). Mean hospital stay and mean convalescence time were 1.7 days (range 1-5 days) and 10 days (range 1-31 days), respectively. Incontinence scores revealed only minor differences between pre- and postoperative values. CONCLUSIONS: SH represents a safe and effective new treatment modality for symptomatic haemorrhoids. Meticulous surgical technique and experience are mandatory to achieve excellent clinical results, e.g., reduced postoperative pain, shortened hospital stay and convalescence. We adopted SH to our surgical armamentarium for the treatment of haemorrhoids grade III and recurrent haemorrhoids. PMID- 11883346 TI - Castleman's disease masquerading as sigmoid colon tumor and Hodgkin lymphoma. AB - Castleman's disease is a benign lymphoid neoplasm first reported as hyperplasia of mediastinal lymph nodes. Some authors referred to the lesions as isolated tumors, described as a variant of Hodgkin's disease with a possibility of a malignant potential and others proposed that the lymphoid masses were of a hamartomatous nature. Three histologic variants and two clinical types of the disease have been described. The disease may occur in almost any area in which lymph nodes are normally found. The most common locations are thorax (63%), abdomen (11%) and axilla (4%). We report two separate histologic types of Castleman's disease which were rare in the literature, mimicking sigmoid colon tumor and Hodgkin lymphoma. The diagnostic and therapeutic aspects of this rare entity is discussed. PMID- 11883347 TI - Manual removal of the placenta and postcesarean endometritis. AB - OBJECTIVE: To determine if manual vs. spontaneous delivery of the placenta at cesarean section affects the rate of postoperative endometritis and amount of blood loss. STUDY DESIGN: A prospective, randomized study was carried out on patients who had cesarean delivery assigned either to spontaneous delivery of the placenta (group 1) or manual removal (group 2). We excluded patients undergoing emergency cesareans and those with possible placenta accreta or evidence of preexisting infection. Outcome measures (frequency of endometritis and quantitative decrease in hemoglobin) were compared for the two groups. RESULTS: Study criteria were met for 375 subjects: 177 in group 1 and 198 in group 2. Endometritis was diagnosed in 1.7% of the former and 2.5% of the latter. The change in hemoglobin, reflecting operative blood loss, was similar in both groups (-1.81 and -1.72 g/dL, respectively). CONCLUSION: We found no significant difference in either postoperative endometritis or blood loss regardless of the means used to effect delivery of the placenta. The frequency of febrile morbidity in our study cases was considerably lower than heretofore reported. PMID- 11883348 TI - Use of dynamic contrast-enhanced MRI to assess the functional vascular pharmacokinetic parameters of normal human ovaries. AB - OBJECTIVE: To assess the ability of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to measure functional vascular pharmacokinetic parameters of normal human ovaries in vivo using an open linear two-compartment pharmacokinetic model. STUDY DESIGN: Twenty-one women with no ovarian disease underwent DCE-MRI. Sequential images were acquired during injection of a contrast bolus. Ovarian volumes were calculated and pharmacokinetic data analyzed using a pharmacokinetic model. Ovarian tissue was compared with skeletal muscle to demonstrate the pharmacokinetic parameters. RESULTS: Normal ovarian tissue was found to enhance rapidly and dramatically. When compared with skeletal muscle as a control, ovarian tissue was seen to demonstrate a significantly higher maximum enhancement factor and amplitude of the upslope, although there was no significant difference in the exchange rate. Sample size precluded conclusions about differences due to menstrual cycle or menopausal status. CONCLUSION: The results reflect the vascular physiology of the normal ovary. Definition of the pharmacokinetic properties of normal ovaries will allow DCE-MRI to be applied prospectively to conduct noninvasive, in vivo studies of ovarian angiogenic function, including response to drugs, contraceptive research, assessment of polycystic ovary syndrome, ovarian hyperstimulation syndrome, diagnosis of malignancy and prediction of response to antiangiogenic chemotherapy. PMID- 11883349 TI - A randomized trial of vaginal prostaglandin E2 for induction of labor. Insert vs. tablet. AB - OBJECTIVE: To compare the efficacy and safety of a prostaglandin E2 (PGE2) vaginal insert with PGE2 administered as a vaginal tablet. STUDY DESIGN: A randomized, observational study was performed. Women requiring induction of labor were randomly assigned to receive either a 10-mg PGE2 vaginal insert (group 1, n = 100) or 3-mg PGE2 tablets twice at six-hour intervals (group 2, n = 100). The primary efficacy outcome variable was vaginal delivery within 24 hours of insertion. The criteria for safety were the occurrence of uterine hyperstimulation, abnormal fetal heart rate patterns, use of beta 2 sympathomimetic drugs and fetal outcome. RESULTS: No differences in terms of vaginal delivery or cesarean section within 24 hours of induction were found. The cesarean section rate was 21% in group 1 and 22% in group 2. The interval from insertion of the induction agent to the onset of regular uterine contractions and the insertion-to-delivery interval were not different between the two cohorts. No difference in the frequency of uterine hyperstimulation, use of beta 2 sympathomimetic drugs, abnormal fetal heart rate patterns, fetal outcome, or oxytocin and analgesic requirements were found. In seven of eight patients in group 1 who experienced uterine hyperstimulation, removal of the insert was sufficient to stop it, whereas in group 2, of nine cases, eight needed medical interventions to end hyperstimulation (P = .003). CONCLUSION: The continuous release of PGE2 from the vaginal insert permits controlled induction of labor, and easy removal of the drug in cases of uterine hyperstimulation is possible. PMID- 11883350 TI - U.S. national trends in labor induction, 1989-1998. AB - OBJECTIVE: To examine the epidemiology of labor induction in the United States. STUDY DESIGN: We used U.S. natality data from 1989 to 1998 and examined the rate of labor induction by year, geographic region, maternal characteristics and pregnancy complications. RESULTS: Between 1990 and 1998, the rate of labor induction increased from 9.5% to 19.4% of all births nationwide. However, the induction rate varied widely by state. White race, higher education and early initiation of prenatal care were associated with a higher rate of induction. For all gestational ages, a significantly increased induction rate occurred during the study period. The increase for clinically indicated induction was significantly slower than the overall increase, suggesting that elective induction has risen much more rapidly. CONCLUSION: The rate of induction of labor more than doubled in the U.S. nationwide in the decade from 1989 to 1998. The increased use of labor induction may be attributable to both clinically indicated and elective induction. PMID- 11883351 TI - Delayed-interval delivery in twin pregnancies. AB - OBJECTIVE: To report on pregnancy outcome in six twin pregnancies with delayed interval delivery in a single maternal-fetal medicine practice. STUDY DESIGN: All cases of attempted delayed-interval delivery from January 1988 to August 2000 in a single maternal-fetal medicine practice were retrospectively reviewed. Patients were managed with a treatment protocol that included rescue cerclage after delivery of the first born twin, antibiotics, corticosteroids and tocolysis. RESULTS: Five of the six twin gestations resulted in viable birth of the second born twin. One pregnancy had loss of both fetuses before viability. All first born twins were nonviable. The median pregnancy prolongation achieved following delivery of the first-born, nonviable twin was 93 days, with a range of 23-153. Three of the five viable, second-born twins had a neonatal intensive care nursery stay of 3, 4 and 35 days (mean, 8.4). No infant required a ventilator. CONCLUSION: Based on our analysis of these six cases, the pregnancy prolongation gained resulted in a clinically significant benefit to the second-born twin, without significant morbidity in the mother. PMID- 11883352 TI - Ovarian function and regulation of the hypothalamic-pituitary-ovarian axis after tubal sterilization. AB - OBJECTIVE: To assess the effects of laparoscopic tubal sterilization with Hulka or Filshie clip on ovarian function and regulation of the hypothalamic-pituitary ovarian axis. STUDY DESIGN: Hormonal changes were evaluated in 33 women undergoing sterilization with Hulka (n = 16) or Filshie clips (n = 17). All participants were healthy, with regular menstrual cycles. The levels of estradiol, follicle-stimulating hormone, luteinizing hormone, sex hormone binding globulin, prolactin, testosterone and androstenedione were measured in one cycle immediately before and 3 and 12 months after sterilization on cycle days 3-7 and 20-24. Repeated measures analysis of variance, paired t test and nonparametric Friedman two-way analysis of variance were used for statistical analysis. RESULTS: The follicular phase estradiol values increased after sterilization. The highest values were observed three months after the procedure (204.8 +/- 119.1 pmol/L vs. 170.3 +/- 111.7 pmol/L) (P = .0407). The values declined to the presterilization level by 12 months (150.3 +/- 71.3 pmol/L). The luteal phase estradiol values did not change significantly. No change in any of the other hormones studied took place, with the exception of a slight increase in follicular phase luteinizing hormone levels (4.4 +/- 1.4 U/L in the first cycle, 5.1 +/- 1.3 U/L in the second cycle and 5.2 +/- 1.8 U/L in the third cycle) (P = .0553). CONCLUSION: Laparoscopic tubal sterilization increases follicular phase estradiol levels, but the change seems to be only temporary. PMID- 11883353 TI - Clinical and endocrine effects of ovulation induction with FSH and hCG supplementation in low responders in the midfollicular phase. A pilot study. AB - OBJECTIVE: To assess the endocrine and clinical effects of luteinizing hormone (LH) activity supplementation administered in the midfollicular phase during controlled ovarian hyperstimulation to poor responders who were candidates for in vitro fertilization (IVF)--embryo transfer. STUDY DESIGN: Prospective, controlled, nonrandomized trial with historical controls. Twenty-five IVF patients who had shown a poor response to standard, long-protocol GnRH-a and FSH only in a preceding cycle (cycle A), were stimulated in the next cycle after six months with hCG supplementation (50 I.U. subcutaneously daily) starting on day 7 during standard, long-protocol GnRH-a and FSH (cycle B). The comparative analysis of clinical effects (duration of stimulation, total highly purified (HP)-FSH dose, number of oocytes retrieved and pregnancy rate) and endocrine responses (serum E2, follicular E2 and androstenedione levels) were determined between cycles A and B. RESULTS: Maximum serum E2 levels and clinical pregnancy rate were higher in cycle B, with hCG supplementation. Also, the follicular E2 and androstenedione levels were higher in cycle B. No differences were noted between cycles as regards the duration of stimulation, total HP-FSH dose and number of oocytes retrieved. CONCLUSION: LH activity supplementation in the midfollicular phase yields favorable pregnancy results in low responders. This may be due to enhanced release of follicular precursors for greater synthesis of E2. PMID- 11883354 TI - Intramuscular methotrexate for tubal pregnancy. AB - OBJECTIVE: To evaluate the safety, convenience and effectiveness of medical treatment of hemodynamically stable tubal pregnancy using intramuscular methotrexate even with adnexal masses up to 5 cm in diameter. STUDY DESIGN: A prospective, observational study was performed on patients admitted with hemodynamically stable tubal pregnancy to the Fourth Unit, Department of Obstetrics and Gynecology, Ain Shams University Maternity Hospital, from September 1999 to August 2000, and fulfilling the inclusion and exclusion criteria. All were given intramuscular methotrexate, 50 mg/m2. RESULTS: Thirty five (66.04%) of 53 patients admitted with tubal pregnancy were eligible for the study. Seventeen patients (48.5%) had an adnexal mass 3.6-5.0 cm in diameter by transvaginal ultrasonography. Thirty-three patients (94.3%) were cured, with 25 (75.8%) requiring a single dose of methotrexate, 7 (21.2%) two doses and 1 (3%) three doses. Two treatment failures (5.7%) occurred, and both had an adnexal mass > 3.5 cm. The mean time for the hCG level to return to normal was 34.8 days (range, 15-70). Of the 33 patients cured, 20 (60.6%) were treated on an outpatient basis, 7 (21.2%) needed brief readmission due to severe separation pain, and 6 (18.2%) were hospitalized for logistic reasons. There was a large statistically significant difference in the serum hCG level when a cutoff level of 1,000 mIU/mL was used to compare those with an adnexal mass < or = 3.5 cm and those with a mass 3.6-5.0 cm as well as those needing more than one dose and those needing one (P < .001). Similarly, there was a statistically significant difference in the number of doses needed between those with an adnexal mass < or = 3.5 cm and those with a mass 3.6-5.0 cm (P < .05). However, multivariate analysis failed to show any statistically significant relation between treatment failure, hCG level, mass size, gestational age, or number of doses due to small sample size and limited number of events. CONCLUSION: Intramuscular methotrexate for hemodynamically stable tubal pregnancy, even in cases with adnexal masses up to 5 cm in diameter, is safe and effective. Larger trials are needed to validate this approach. PMID- 11883355 TI - Rudimentary uterine horn pregnancy. The 20th-century worldwide experience of 588 cases. AB - OBJECTIVE: To investigate the outcomes of rudimentary uterine horn pregnancies and to identify trends and opportunities for improvement in patient care. STUDY DESIGN: During the period 1900-1999, 588 cases of rudimentary uterine horn pregnancy were identified using both manual and computerized searches of Index Medicus, Excerpta Medica and the Index-Catalogue of the Library of the Surgeon General's Office of the United States Army as well as standard reference tracing. Nine characteristics of each case were evaluated: (1) fetal status at birth, (2) maternal survival, (3) neonatal survival, (4) gestational age at delivery, (5) whether the rudimentary horn ruptured, (6) communication status of the horn with the contralateral hemiuterus, (7) gravidity and parity, (8) side of the horn, and (9) order of the gestation. RESULTS: Newborn survival ranged from 0-13% by decade and trended upward. Eighty-five percent of pregnancies occupied noncommunicating horns. Thirty percent of gestations progressed to term or beyond. Fifty percent of pregnant uterine horns ruptured, with 80% of these events occurring before the third trimester. There was no trend in either the incidence or timing of uterine horn rupture during the century. Maternal mortality decreased from 6% to 23% during the first half of the century to < 0.5% currently. Twin pregnancies consisted of 5.3% of cases. CONCLUSION: Neonatal survival has improved greatly for rudimentary horn pregnancies, and maternal mortality has decreased significantly. Such pregnancies are now identifiable early in gestation by obstetric imaging studies, and there can be guarded optimism that favorable trends in outcomes will continue into the 21st century. PMID- 11883356 TI - Recurrent pelvic hydatid cyst obstructing labor, with a concomitant hepatic primary. A case report. AB - BACKGROUND: Hydatid disease in pregnancy is a rare condition, with an incidence of 1/20,000 pregnancies. Although the female reproductive system is a rare site for hydatid disease, various obstetric and gynecologic presentations have been reported. CASE: A 31-year-old multipara was evaluated due to obstructed labor. Sonographic evaluation revealed an 18-cm hepatic and 15-cm pelvic hydatid cyst at 38 weeks of gestation. A healthy, 3,200-g infant was delivered by cesarean section. The cyst, originating in the right ovary and occupying the pouch of Douglas, was removed surgically. The hepatic cyst was decompressed via the percutaneous approach. The follow-up was uneventful. CONCLUSION: Hydatid disease should be considered in the differential diagnosis of adnexal masses in pregnancy. The percutaneous technique is a promising treatment option for hepatic cysts. Not only the diagnosis but also the treatment of hydatid disease is facilitated with currently available magnetic resonance imaging, computed tomography and ultrasonography. PMID- 11883357 TI - Prolonged-interval delivery between the first and second twin. A case report. AB - BACKGROUND: Prolonged-interval delivery between twins can improve neonatal outcome and, under careful monitoring, poses minimal maternal risk. CASE: A 27 year-old, nulliparous woman conceived after in vitro fertilization and was found to have diamniotic-dichorionic twins. At 17 weeks she presented with premature preterm rupture of the membranes of twin A. She was offered delivery or expectant management. She chose expectant management and was discharged. At 18 weeks she delivered twin A and decided to expectantly manage the second twin. Amniocentesis was performed to evaluate for intraamniotic infections. There was no evidence of them, and a McDonald cerclage was placed. At 32 weeks, spontaneous rupture of the membranes occurred for twin B. The patient delivered vaginally a male infant (2,070 g) who did not need mechanical ventilation and was discharged from neonatal intensive care on the 7th day of life, with no complications. CONCLUSION: Expectant management of a second twin after delivery of the first in selected patients can improve neonatal outcome. PMID- 11883358 TI - Entrapment of viable trophoblastic tissue in a uterine hematoma after surgical evacuation. A case report. AB - BACKGROUND: Postevacuation uterine perforation is a common event. Early diagnosis and management are important to minimize the associated morbidity and mortality. CASE: A woman presented with persistent vaginal bleeding for two weeks following surgical uterine evacuation for missed abortion at 7 weeks' gestation. She had a persistently elevated serum human chorionic gonadotropin level. Ultrasonography revealed a 3-cm, heterogeneous mass with high vascularity at the left anterior uterine fundal region; the endometrial echo was normal. Cornual pregnancy was suspected, and surgical resection was planned. Intraoperatively, a uterine hematoma with evidence of previous uterine perforation was diagnosed. Hysterotomy, removal of the hematoma and repair of the uterus were performed. Histologic examination revealed entrapment of trophoblastic tissue in the specimen. The patient had an uneventful recovery. CONCLUSION: Entrapment of trophoblastic tissue in a uterine hematoma is a rare sequel of uterine perforation after evacuation and might be confused with cornual pregnancy. PMID- 11883359 TI - Office microlaparoscopy for female sterilization under local anesthesia. A cost and clinical analysis. AB - OBJECTIVE: To evaluate the clinical and cost effectiveness of permanent female sterilization using microlaparoscopy and bipolar cauterization. The authors also tested how well the procedure was tolerated when performed under local anesthesia and conscious sedation. STUDY DESIGN: Patients undergoing microlaparoscopic permanent sterilization were prospectively followed. The rate of procedure completion, patients' pain scores and complication rates were reviewed. RESULTS: The procedure was completed in 93% of patients (28 of 30). The pain scores decreased substantially within one day after the procedure. The total cost savings for the 29 cases were $16,211. Two patients experienced severe intraoperative discomfort and were rescheduled for endoscopic sterilization under general anesthesia. There were no complications. CONCLUSION: Permanent female sterilization using microlaparoscopy is cost effective and safe in selected patients, patient exclusion criteria need to be instituted to ensure the safety and success of the procedure. PMID- 11883360 TI - [Internet communication between family physicians and the university hospital]. AB - The potential of electronic communication in medicine is assessed based on an analysis of a pilot project pertaining to internet based communication among referring and hospital physicians. Advantages of electronic data exchange in medicine pertain to speed and capacity for data transfer, availability of data and data integration, ultimately enabling consistent medical case management. Quality requirements of electronic communication of medical data are related to safety, availability, data integration, potential for case management and system qualities. Medical efficiency can be increased by use of electronic communication only if complex functions beyond the substitution of conventional mail by e-mail are implemented and an exhaustive use of the technology can be achieved. PMID- 11883361 TI - [Pregnancy outcome in 1,252 fetuses after nuchal translucency measurement in the 1st trimester]. AB - The measurement of the nuchal translucency is an important marker to detect a vast number of fetal anomalies. Lately the nuchal translucency has been used increasingly as a screening method to find chromosomal anomalies especially for trisomy 21. Beside a high incidence of chromosomal defects one assumes a high risk of rare syndromes and other associated anomalies such as heart defects, skeletal anomalies, cerebral anomalies, diaphragmatic hernias, as well as of an intrauterine death. In view of abortion rate, detection rate of chromosomal anomalies, other fetal anomalies and rare syndromes we evaluated in this study the pregnancy outcome after nuchal translucency measurement, and discuss a concept for its management. PMID- 11883362 TI - [Acute fatty liver in pregnancy: clinical and histopathological course. Case report]. AB - Acute fatty liver of pregnancy is a rare disease which may be letal if diagnosis is missed. The pathogenesis is not completely clear, but there is some evidence that some cases have been associated with a genetic deficiency of fatty acid beta oxidation. Other predisposing factors include primiparity, multiple pregnancy, male fetal sex and pre-eclampsia. Clinical presentation and laboratory findings are often unspecific. Increasing serum aminotransferases are characteristic in the early stage of the disease. Liver biopsy establishes the diagnosis and typically shows microvesicular, centrilobular fatty changes of hepatocytes. Differential diagnosis includes the HELLP-Syndrome, cholestasis of pregnancy, pre eclampsia and viral or drug induced hepatitis. Without adequate treatment liver failure with coagulopathy and encephalopathy may develop. Two cases of acute fatty liver in pregnancy in an early stage are presented. Clinical and histopathological findings as well as diagnostic and therapeutic procedures are discussed. PMID- 11883363 TI - [Effect of new technologies on diagnosis and therapy of acute appendicitis]. AB - Acute appendicitis remains a diagnosis based primarily on the history and the physical examination performed by an experienced surgeon. Ultrasonography and CT can be an useful adjunct, but they should not be used without context to the clinical picture. In therapy, open appendicectomy remains the golden standard. Laparoscopic appendectomy is reserved for special situations. Preoperative ultrasound is useful to decide upon the operative procedure. When the clinical picture and ultrasonography reveals acute appendicitis one can expect a minimal negative appendectomy- and laparotomy rate of 2.7% and 2.1%. In these situations open appendectomy is indicated. In the case where ultrasonography reveals no appendicitis, negative appendectomy rate is 31%. In this situation further abdominal exploration and thus diagnostic laparoscopy and laparoscopic appendectomy is indicated. PMID- 11883364 TI - [88-year-old patient with known aortic stenosis and new onset gastrointestinal hemorrhage]. PMID- 11883365 TI - [Ion channel abnormalities ("channelopathies") in neurologic diseases]. AB - THE ROLE OF IONIC CHANNEL DYSFUNCTION: During various neurological diseases has been evoked for many years on electro-physiological data. Molecular biology has led to great progress in neurology, and can be considered "functional" since it is surpasses the classical anatomo-clinical methods. IONIC CHANNEL DYSFUNCTION: Can be determined genetically, resulting from the mutation of a gene code of a channel sub-unit. CHANNELOPATHIES ARE RESPONSIBLE: For muscular diseases (myotonia, familial periodic paralysis, malignant hyperthermia and congenital myasthenia), but also for central nervous system disorders such as familial hemiplegic migraine, hereditary paroxystic ataxia and certain forms of Mendel's law hereditary epilepsy. ACQUIRED IONIC CHANNEL DYSFUNCTION: Resulting from auto immune aggression is implied in diseases such as Lambert-Eaton's myasthenic syndrome and Isaac's neuromyotonia syndrome. It probably plays a part in the clinical, and particularly the sensitive expression (paresthesia and pain) of some peripheral neuropathies and certain central nervous system affections, such as multiple sclerosis. PMID- 11883367 TI - [Acute viral meningitis. Epidemiologic, clinical and biological characteristics of 29 hospitalized patients]. PMID- 11883366 TI - [Evaluation of the risk of abortion abuse resulting from the two-week legal delay in France]. AB - OBJECTIVE: The legal delay for abortion in France has recently been prolonged from 10 (12 weeks of amenorrhea) to 12 weeks (14 weeks of amenorrhea). With the progress in sonography, certain foetuses may exhibit malformations during the first trimester. Diagnosis of foetal gender at 12 weeks of amenorrhea is sometimes possible. We studied the possibility that the prolonged legal delay before abortion might incite women to abort, simply on sonographic criteria. METHODS: Our enquiry was conducted in March 2001 in the department of Obstetrics and Gynaecology of the Jean Verdier hospital in Bondy. Two questionnaires were drawn-up by the Medical Ethics Laboratory of the Necker Hospital in Paris and were distributed to 128 women and 24 nurses. RESULTS: Recourse to abortion was high if laparoschisis or the absence of a hand was revealed, low in the case of opacity of the neck, and almost inexistent in the case of unwanted gender, in a female population with less than one child. CONCLUSION: A national consensus on foetal abnormalities to be searched for, and not to be searched for (Number of fingers? Upper lip?...) and the eventual detection of the gender during sonography of the 1st trimester is urgent in view of the technological progress made. Clear and reassuring information is essential when confronted with foetal abnormalities and must lead to complete and precise antenatal diagnosis (caryotyping, sonographic control 2 or 3 weeks later). PMID- 11883368 TI - [Atypical cutaneous leishmaniasis]. AB - French Guyana is an endemic area for American cutaneous leishmaniasis. At the start, the initial red lesion may be mistaken for a whitlow. Twenty percent of developed forms exhibit a sporotrichosis pattern. The notion of travel to an endemic area is very important for diagnosis. We report the case of a young man from French Guyana presenting with a cutaneous lesion of the finger. After numerous surgical treatments for a "whitlow", the final diagnosis of leishmaniasis was difficult because of local-complications and cutaneous rearrangement. Erroneous initial orientation in a unit unaware of tropical diseases can forestall appropriate care of the patients. Epidemiological, clinical and therapeutic data on American tegument leishmaniasis are discussed. PMID- 11883369 TI - [Mycobacterium kansasii infection in an immunocompetent patient with a healthy pulmonary parenchyma]. PMID- 11883370 TI - [Treatment with F(ab')2(Viperfav) for viper envenomation]. PMID- 11883371 TI - [Mucoviscidosis in adults]. AB - FROM CHILDREN TO ADULTS: Mucoviscidosis, a genetic autosomal recessive disease, is not only a paediatric disease, but with the progress in therapy, has become a disease of adults. Today, median survival of patients is of 30 years and, in France, more than one third of patients are adults. CLINICAL SYMPTOMS IN ADULTS: Are predominantly respiratory, with dilatation of the bronchi and characteristic colonization flora. Chronic bronchial Pseudomonas aeruginosa colonisation is frequently encountered in adults. It develops in successive episodes towards chronic respiratory failure. Other than this typical form, diagnosed in the first years of life, the discovery of the CFTR (Cystic Fibrosis Transmembrane conductance Regulator) gene and its mutations permits diagnosis of mucoviscidosis in patients presenting with mild, or even monosymptomatic forms of the disease. Today, diagnosis of mucoviscidosis relies on the association of characteristic organ damage and an abnormality in CFTR (sweat test and/or difference in nasal potential) or the revelation of gene mutations on each allele. REGARDING TREATMENT: Respiratory failure is the core of daily therapeutic efforts. Respiratory physical therapy, effort re-education and muscle exercising are essential. Antibiotherapy is aimed at treating, spacing out or preventing the infectious exacerbations, in order to stall the functional degradation. Repeated, sequential cycles of intravenous infusions of antibiotics are required in P. aeruginosa chronic bronchial colonization. Treatment of the primary pyocyanic colonisation and inhaled antiobiotherapy appear promising. Pulmonary transplantation is a recognized and efficient therapeutic in advanced stages of respiratory failure. The discomfort and time the patient has to spend on daily treatments requires regular monitoring, to improve compliance and to improve the quality of life of these patients. PMID- 11883373 TI - Abstracts of the meeting of the Israel Society for Auditory Research. Tel Aviv, Israel, October 16, 2001. PMID- 11883372 TI - [The hepatopulmonary syndrome]. AB - DEFINITION: The hepatopulmonary syndrome (HPS) associates a chronic hepatic affection, hypoxemia < 70 mm Hg and pulmonary vasodilatation. PHYSIOPATHOLOGY: The mechanisms leading to pulmonary vasodilatation are complex and unclear. There appears to be an imbalance between the vasodilatating and vasoconstricting mediators. Nitrogen monoxide and endotheline-1 are well known. Hypoxia can be explained by the association of heterogenic ventilation-perfusion, shunts (rare), and a default in "diffusion-perfusion". CLINICAL ASPECTS: In a hypoxic patient, platypnoea and orthodeoxia are characteristic of HPS. Stellar angioma associated with digital hippocratism and signs of portal hypertension are usually present. TO PERMIT DIAGNOSIS: The air of blood gases, followed by 100% O2, standing and reclining, must be measured in all cirrhotic patients to detect hypoxemia. Contract sonography is the key diagnostic examination. Pulmonary perfusion scintigraphy establishes prognosis. Pulmonary angiography differentiates two groups of patients and, for type II patients, embolization therapy can be proposed. Preliminary data indicate that densitometry, conducted in rigorous conditions, can show pulmonary vasodilatation. Its interest must be confirmed by further studies on larger cohorts of patients. THERAPEUTIC POSSIBILITIES: The only efficient treatment of HPS is hepatic transplant (HT). The placing of an intra-hepatic portal systemic shunt can be proposed while waiting for HT, or in certain patients not requiring HT. No medical treatment has demonstrated its efficacy, but better knowledge of the physio-pathologic mechanisms should improve this situation in the future. PMID- 11883374 TI - Paradoxical air embolism during hepatic resection. AB - Systemic venous air embolism is a serious complication in patients with chronic liver disease having liver surgery. Intrapulmonary arteriovenous shunting can permit air emboli to pass into the systemic circulation. We describe a case of paradoxical air embolism detected by transoesophageal echocardiography in a patient with cirrhosis who was having a hepatic resection. PMID- 11883375 TI - Effects of high inspired oxygen fraction during elective caesarean section under spinal anaesthesia on maternal and fetal oxygenation and lipid peroxidation. AB - BACKGROUND: Oxygen supplementation is given routinely to parturients undergoing Caesarean section under regional anaesthesia. While the aim is to improve fetal oxygenation, inspiring a high oxygen fraction (FIO2) can also increase free radical activity and lipid peroxidation in both the mother and baby. In this prospective, randomized, double-blind study, we investigated the effect of high inspired oxygen fraction (FIO2) on maternal and fetal oxygenation and oxygen free radical activity in parturients having Caesarean section under spinal anaesthesia. METHODS: Forty-four healthy parturients were randomized to breathe either 21% (air group) or 60% oxygen (oxygen group) intraoperatively via a ventimask. Maternal arterial blood was collected at 5-min intervals from baseline until delivery, and umbilical arterial and venous blood was collected at delivery. We measured blood gases and the products of lipid peroxidation (8 isoprostane, malondialdehyde (MDA), hydroperoxide (OHP)) and purine metabolites. RESULTS: At delivery, the oxygen group had greater maternal arterial PO2 [mean 30.0 (SD 6.3) vs 14.2 (1.9) kPa; mean difference 15.8 kPa, 95% confidence interval 12.9-18.7 kPa, P<0.001] and greater umbilical venous PO2 [4.8 (1.0) vs 4.0 (1.4) kPa; mean difference 0.8 kPa, 95% confidence interval 0.0-1.5 kPa, P=0.04] compared with the air group. Maternal and umbilical plasma concentrations of lipid peroxides (8-isoprostane, MDA, OHP) were greater in the oxygen group than in the air group (P<0.05). CONCLUSIONS: We conclude that breathing high FIO2 modestly increased fetal oxygenation but caused a concomitant increase in oxygen free radical activity in both mother and fetus. PMID- 11883376 TI - Model-based administration of inhalation anaesthesia. 3. Validating the system model. AB - BACKGROUND: We quantified the predictive performance of our computer model of the administration of inhalation anaesthesia from a Datex-Ohmeda Modulus CD circle absorber system. METHODS: In 50 patients, desflurane anaesthesia was maintained with a fresh-gas flow (FGF) of 0.5 litres min(-1) of both nitrous oxide and oxygen, preceded by fast (n=14) or slow (n=36) induction: FGF greater than total ventilation, Group F; FGF equal to 1.0 litres min(-1), Group S. The two versions of the model studied differed in the size of their inter-tissue diffusion, as 0 (version 1) and 3% (version 2) of the cardiac output was shifted from the viscera to adipose tissue. Model performance was judged by comparing measured and predicted gas concentrations in terms of three variables for each gas concentration in each patient: root mean squared error (rmse=total error), bias (mean predicted - measured) (systematic error), and scatter (error around bias). These variables were then averaged over all patients. These measures were calculated overall, and separately for each group and each stage (1 = induction or 2 = maintenance). RESULTS: Model predictions were in reasonable to very good agreement with clinically obtained data. Version 2 performed better than version 1. Differences between groups were not demonstrated. The model performed better for stage 2, but only for desflurane. In group S, results (mean (SD); as percentages of the measured values for nitrous oxide, oxygen and desflurane) in the order rmse, bias, and scatter were for end-tidal concentrations of nitrous oxide: 8 (4), 8 (5), 2 (1)%; oxygen: 11 (4), -10 (6), 2 (1.1)%; nitrogen: 0.9 (0.6), -0.8 (0.6), 0.2 (0.1) vol%; carbon dioxide: 1.8 (0.6), 1.8 (0.6), 0.2 (0.1) vol%; desflurane, stage 2: 8 (4), 4 (7), 4 (2)%, vs 15 (6), -10 (8), 9 (4)% for stage 1. CONCLUSION: Administration of inhalation anaesthesia can be based on version 2 of this model, but must be guided by active monitoring. PMID- 11883377 TI - Predictive performance of a physiological model for enflurane closed-circuit anaesthesia: effects of continuous cardiac output measurements and age-related solubility data. AB - BACKGROUND: The disposition of inhalation anaesthetics is governed by the factors described in the Fick principle. METHODS: We have recalibrated a previously validated physiological model for enflurane closed-circuit inhalation anaesthesia, using individual continuous cardiac output measurements as well as age-related enflurane solubility coefficients as inputs to the model. Two model versions using 'calculated' (Brody's formula) or 'measured' (thoracic electrical bioimpedance) cardiac output values, and two versions with 'standard' (fixed) or 'age-related' solubility coefficients were formulated. RESULTS: Data from 62 ophthalmic surgical patients were used to validate the predictive performance of the four model versions. The root mean squared errors (total error) and scatters (error variation) were similar with the extended model versions, but the group biases (systematic error component) were significantly less with the model versions that included age-related solubility compared with the versions using standard solubility coefficients (bias -0.76/-0.78% vs -3.44/-3.60%). CONCLUSION: The inclusion of age-related solubility coefficients but not of continuous cardiac output measurements improves the predictive performance of the physiological model for closed-circuit inhalation anaesthetic conditions in routine clinical practice. PMID- 11883378 TI - Naloxone prevents increased atrial natriuretic peptide release during regional myocardial ischaemia and stunning in awake dogs. AB - BACKGROUND: Atrial natriuretic peptide (ANP) release is increased in patients with ischaemic left ventricular dysfunction. A beneficial effect of naloxone on recovery from myocardial stunning was shown previously. The aim of this study was to investigate the effects of naloxone on ANP release during regional myocardial ischaemia and stunning in awake dogs. METHODS: Ten dogs were chronically instrumented for measurement of heart rate, left atrial, aortic, and left ventricular pressure (LVP), LV dP x dtmax/min(-1), and myocardial wall-thickening fraction. An occluder around the left anterior descending artery (LAD) allowed induction of reversible ischaemia in the LAD-perfused myocardium. Each dog underwent two ischaemic episodes (randomized crossover fashion; separate days): 10 min of LAD occlusion (1) after application of naloxone (63 microg kg(-1)), and (2) without naloxone. ANP levels were measured at baseline (BL) and at predetermined time points until complete recovery of myocardial stunning occurred. RESULTS: LAD ischaemia-induced release of ANP (peak level: 182 (30) vs 27 (7) pg ml(-1) BL) only in the control group without naloxone. Between 1 and 180 min of reperfusion, ANP levels were significantly higher only in the control group (P<0.05). CONCLUSION: Pre-ischaemic application of naloxone prevents this ischaemia-induced ANP-release in conscious dogs. PMID- 11883379 TI - Growth and development. Web alert. PMID- 11883380 TI - How do we address MD distributions? PMID- 11883381 TI - AHA calls 2002 APC changes 'unworkable'. PMID- 11883382 TI - Hospital underwriting: why not? PMID- 11883383 TI - Achieving our vision. PMID- 11883384 TI - Model-based administration of inhalation anaesthesia. 4. Applying the system model. AB - BACKGROUND: We developed and tested a simple dosing strategy for rapid induction with isoflurane followed by maintenance under minimal-flow conditions, that is 0.5 litre min(-1) total fresh gas flow (FGF). An end-expired concentration was to be achieved within 5 min in a desired therapeutic window, that is 0.8-1.1 vol%, and to be maintained within it for at least 30 min. METHODS: With our new model we computed a three-stage regimen using one fixed vaporizer setting: 3 vol% isoflurane in a FGF of 3 and 1.5 litre min(-1), each for 3 min, and 0.5 litre min thereafter. The ratio of nitrous oxide:oxygen was, consecutively, 2:1, 2:1, and 2:3. We evaluated this scheme in 58 adult patients (body mass 74 (SD 13) kg), mostly during eye and ear, nose, and throat surgery. RESULTS: Measured oxygen (33 45 vol%) and nitrous oxide concentrations (66-50 vol%) evolved in accordance with those computed. In five patients with a median of body mass 92 kg (range 76-126 kg), inspired oxygen concentrations decreased to less than 30 vol%. End-expired isoflurane concentration entered the window after 2 min (range 1.0-5.67 min) and attained its maximum, that is 0.96 vol% (0.8-1.2 vol%), after 3.45 min (1.67-6.33 min). The mean end-expired concentration was in the desired window from 3-60 min and an average of 72% of individual measurements were within the window from 3-30 min. The scheme was adapted in six patients (excluded from analysis) because of hypotension. CONCLUSION: The regimen is easily remembered, reliable, and lends itself to alternative strategies, but must be guided by the monitoring of gas and vapour concentrations and haemodynamic variables. PMID- 11883385 TI - Cardiovascular effects of simultaneous occlusion of the inferior vena cava and aorta in patients treated with hypoxic abdominal perfusion for chemotherapy. AB - BACKGROUND: Animal studies suggest less cardiovascular disturbance if the aorta and vena cava are occluded simultaneously. We set out to establish the effects of simultaneous clamping in humans, because oncologists suggested that perfusion for chemotherapy could be done under local anaesthesia without invasive haemodynamic monitoring. METHODS: We studied the cardiovascular effects of the onset and removal of simultaneous occlusion of the thoracic aorta and inferior vena cava, in seven ASA II patients. Two stop-flow catheters positioned in the aorta and in the inferior vena cava were inflated to allow hypoxic abdominal perfusion to treat pancreatic cancer. We measured the arterial pressure, heart rate (HR), right atrial pressure (RAP), pulmonary artery pressure (PAP), pulmonary artery wedge pressure (PAWP) and cardiac output (CO), and calculated systemic vascular resistance index (SVRi), pulmonary vascular resistance index (PVRi), left ventricular stroke work index (LVSWi) and right ventricular stroke work index (RVSWi). Three patients were studied with transoesophageal echocardiography. RESULTS: Six patients needed intravenous nitroprusside during the occlusion because mean arterial pressure (MAP) increased to more than 20% of baseline (SVRi increased by 87%). One minute after occlusion release, all patients had a 50% decrease in MAP, and mPAP increased by 50%. The procedure had severe cardiovascular effects, shown by a 100% increase in cardiac index at occlusion release with increases in left and right ventricular stroke work indices of 75% and 147%. Left ventricular wall motion abnormalities were seen on transoesophageal echocardiography. CONCLUSIONS: Serious haemodynamic changes occur during simultaneous occlusion of the thoracic aorta and inferior vena cava, which may need invasive haemodynamic monitoring. PMID- 11883387 TI - Comparison of predictive models for postoperative nausea and vomiting. AB - BACKGROUND: In order to identify patients who would benefit from prophylactic amtiemetics, six predictive models have been described for the risk assessment of postoperative nausea and vomiting (PONV). This study compared the validity and practicability of these models in patients undergoing general anaesthesia. METHODS: Data were analysed from 1566 patients who underwent balanced anaesthesia without prophylactic antiemetic treatment for various types of surgery. A systematic literature search identified six predictive models for PONV. These models were compared with respect to validity (discriminating power and calibration characteristics) and practicability. Discriminating power was measured by the area under the receiver operating characteristic curve (AUC) and calibration was assessed by weighted linear regression analysis between predicted and actual incidences of PONV. Practicability was assessed according to the number of factors to be considered for the model (the fewer factors the better), and whether the score could be used in combination with a previously applied cost effective concept. RESULTS: The incidence of PONV was 600/1566 (38.1%). The discriminating power (AUC) obtained by the models (named according to the first author) using the risk classes from the recommended prophylactic concept were as follows: Apfel, 0.68; Koivuranta, 0.66; Sinclair, 0.66; Palazzo, 0.63; Gan, 0.61; Scholz, 0.61. For four models, the following calibration curves (expressed as the slope and the offset) were plotted: Apfel, y=0.82x+0.01, r2=0.995; Koivuranta, y=1.13x-0.10, r2=0.999; Sinclair, y=0.49x+0.29, r2=0.789; Palazzo, y=0.30x+0.30, r2=0.763. The numbers of parameters to be considered were as follows: Apfel, 4; Koivuranta, 5; Palazzo, 5; Scholz, 9; Sinclair, 12; Gan, 14. CONCLUSION: The simplified risk scores provided better discrimination and calibration properties compared with the more complex risk scores. Therefore, simplified risk scores can be recommended for antiemetic strategies in clinical practice as well as for group comparisons in randomized controlled antiemetic trials. PMID- 11883386 TI - Ketorolac, diclofenac, and ketoprofen are equally safe for pain relief after major surgery. AB - BACKGROUND: Ketorolac is approved for the relief of postoperative pain but concerns have been raised over a possible risk of serious adverse effects and death. Two regulatory reviews in Europe on the safety of ketorolac found the data were inconclusive and lacked comparison with other non-steroidal anti inflammatory drugs. The aim of this study was to compare the risk of serious adverse effects with ketorolac vs diclofenac or ketoprofen in adult patients after elective major surgery. METHODS: This prospective, randomized multicentre trial evaluated the risks of death, increased surgical site bleeding, gastrointestinal bleeding, acute renal failure, and allergic reactions, with ketorolac vs diclofenac or ketoprofen administered according to their approved parenteral and oral dose and duration of treatment. Patients were followed for 30 days after surgery. RESULTS: A total of 11,245 patients completed the trial at 49 European hospitals. Of these, 5634 patients received ketorolac and 5611 patients received one of the comparators. 155 patients (1.38%) had a serious adverse outcome, with 19 deaths (0. 17%), 117 patients with surgical site bleeding (1.04%), 12 patients with allergic reactions (0.12%), 10 patients with acute renal failure (0.09%), and four patients with gastrointestinal bleeding (0.04%). There were no differences between ketorolac and ketoprofen or diclofenac. Postoperative anticoagulants increased the risk of surgical site bleeding equally with ketorolac (odds ratio=2.65, 95% CI=1.51-4.67) and the comparators (odds ratio=3.58, 95% CI=1.93-6.70). Other risk factors for serious adverse outcomes were age, ASA score, and some types of surgery (plastic/ear, nose and throat, gynaecology, and urology). CONCLUSION: We conclude that ketorolac is as safe as ketoprofen and diclofenac for the treatment of pain after major surgery. PMID- 11883389 TI - Submental intubation in a patient with beta-thalassaemia major undergoing elective maxillary and mandibular osteotomies. AB - A 33-yr-old woman with marked maxillo-facial deformities as a result of underlying beta-thalassaemia major was to undergo corrective maxillary and mandibular osteotomies. The placement of an endotracheal tube posed a problem in this patient because of anatomical deformities in her nasal passage, surgical constraints on using the oral route, and reluctance of the patient to have a tracheostomy. This case report describes the use of a submental tracheal intubation technique, and the associated anaesthetic difficulties encountered in patients with this pathology. PMID- 11883388 TI - Differential effects of intravenous anaesthetic agents on the response of rat mesenteric microcirculation in vivo after haemorrhage. AB - BACKGROUND: The differential effects of i.v. anaesthesia on the response of the mesenteric microcirculation after haemorrhage in vivo are previously unexplored. METHODS: Male Wistar rats (n=56) were anaesthetized intravenously either with propofol and fentanyl (propofol/fentanyl), ketamine or thiopental. A tracheostomy and carotid cannulation were performed and the mesentery surgically prepared for observation of the microcirculation using fluorescent in vivo microscopy. Animals were allocated to one of three groups: control, haemorrhage or haemorrhage re infusion. RESULTS: After haemorrhage, the response of the microcirculation differed during propofol/fentanyl, ketamine and thiopental anaesthesia. During propofol/fentanyl anaesthesia there was constriction of arterioles (-16.7 (3.9)%), venules (-5.9 (1.7)) and capillaries (-16.3 (2.8)) (n=12). During ketamine and thiopental anaesthesia both constriction and dilation was observed. After haemorrhage and re-infusion, macromolecular leak occurred from venules during propofol/fentanyl and thiopental anaesthesia (P<0.05), but not during ketamine anaesthesia. CONCLUSION: In summary, i.v. anaesthetic agents differentially alter the response of the mesenteric microcirculation to haemorrhage. PMID- 11883391 TI - Blood pressure manipulation during loco-regional anaesthetic carotid surgery. PMID- 11883390 TI - Postoperative recovery after inguinal herniotomy in ex-premature infants and the use of caffeine. PMID- 11883392 TI - Bibliography. Current world literature. Obstructive, occupational and environmental diseases. PMID- 11883393 TI - Exercise echocardiography or exercise SPECT imaging? A meta-analysis of diagnostic test performance. PMID- 11883395 TI - Editor's corner. PMID- 11883394 TI - Giving our little princess her new voice. PMID- 11883396 TI - Innovative breast cancer care program improves patient satisfaction, care. PMID- 11883397 TI - Guidelines help target diagnosis and treatment of common sinus infection. PMID- 11883398 TI - Top hospitals more likely to use stents and clot-inhibiting drugs in angioplasty. PMID- 11883399 TI - OSHA clarifies, updates Bloodborne Pathogens Standard. PMID- 11883400 TI - Spectrophotometric determination of ziram in a commercial sample and wheat by extraction of its copper dimethyldithiocarbamate complex into molten naphthalene. AB - A method was developed for the determination of zinc(II) dimethyldithiocarbamate by converting it into the copper(II) dithiocarbamate complex, which is then extracted into molten naphthalene. The absorbance is measured at 430 nm versus a reagent blank. Beer's law is obeyed for concentrations of 0.63 x 10(-3) to 17.2 x 10(-3) g/L in the final solution. The method is sensitive and was applied to the determination of ziram in a commercial sample and in wheat grain. PMID- 11883402 TI - National spotlight on children's mental health. PMID- 11883401 TI - Determination of vitamin B12 in milk products and selected foods by optical biosensor protein-binding assay: method comparison. AB - Biomolecular interaction analysis was evaluated for the automated determination of vitamin B12 in a range of foods. The analytical technique was configured as a biosensor-based, nonlabeled inhibition protein-binding assay using nonintrinsic R protein. Sample extraction conditions were optimized, and both ligand specificity and nonspecific binding considerations were evaluated. Performance parameters included a quantitation range of 0.08-2.40 ng/mL, recoveries of 89-106%, agreement against assigned reference values for 3 independent certified food reference materials, and a mean between-laboratory reproducibility relative standard deviation of 4.9%. The proposed method was compared with reference microbiological and radioisotope protein-binding methods for a range of food samples. A wide selection of milks, infant formulas, meats, and liver were evaluated for their vitamin B12 content. The influence of season was studied in herd milk, early lactation was followed for a single animal, and the cobalamin content of bovine, caprine, and ovine milks was compared. PMID- 11883403 TI - Post-traumatic stress disorder in children: controversies and unresolved issues. AB - TOPIC: Questions regarding how severely traumatized children may meet diagnostic criteria for an accurate diagnosis of posttraumatic stress disorder (PTSD), the comorbidity of PTSD with many other psychiatric illnesses, and the possibility that PTSD is not a valid diagnostic formulation as it has been applied to children and adolescents demonstrate that the currently held concept of PTSD may not be operationally sound. PURPOSE: To explore recent empirical studies to demonstrate the current state of controversy and postulate future direction of the conceptual framework. SOURCES: Selected published literature. CONCLUSIONS: PTSD in children and adolescents, as it is popularly understood as a conceptual framework, is found to be undergoing a conceptual metamorphosis. PMID- 11883404 TI - Seclusion and restraint of children: a literature review. AB - TOPIC: Nurses' attitudes toward the use of seclusion and restraint with children. PURPOSE: To review recent literature concerning these controversial interventions, and to examine possible alternative therapeutic interventions. SOURCES: Selected published nursing and psychiatric literature 1987 to 1998. CONCLUSIONS: Staff have a positive attitude toward the use of seclusion and restraint. A theory of power and control may explain their use when many alternative, less restrictive interventions are available. PMID- 11883405 TI - That place where we live: the discovery of self through the creative play experience. AB - TOPIC: The origin and meaning of play in relation to D.W. Winnicott's (1971) conceptualization of the intermediate area of experience and transitional object, and transitional phenomena formation. PURPOSE: To demonstrate how the nurse therapist can enhance the child's discovery of self and healthy development through creativity and play. SOURCES: Clinical experiences with children in a school-based mental health program and literature support. CONCLUSIONS: By providing "good enough" mothering and a holding environment to enhance the development of a creative play experience, the nurse allows the child to find meaning through self-expression and, ultimately, health. PMID- 11883406 TI - Eye on Washington: cultural competence: implications of the Surgeon General's report on mental health. PMID- 11883407 TI - ACAPN (division of ISPN) position statement on the rights of children in treatment settings. PMID- 11883408 TI - Improving risk assessment: hip geometry, bone mineral distribution and bone strength in hip fracture cases and controls. The EPOS study. European Prospective Osteoporosis Study. AB - Hip geometry and bone mineral density (BMD) have previously been shown to relate independently to hip fracture risk. Our objective was to determine by how much hip geometric data improved the identification of hip fracture. Lunar pencil beam scans of the proximal femur were obtained. Geometric and densitometric values from 800 female controls aged 60 years or more (from population samples which were participants in the European Prospective Osteoporosis Study, EPOS) were compared with data from 68 female hip fracture patients aged over 60 years who were scanned within 4 weeks of a contralateral hip fracture. We used Lunar DPX 'beta' versions of hip strength analysis (HSA) and hip axis length (HAL) applied to DPX(L) data. Compressive stress (Cstress), calculated by the HSA software to occur as a result of a typical fall on the greater trochanter, HAL, body mass index (BMI: weight/(height)2) and age were considered alongside femoral neck BMD (FN-BMD, g/cm2) as potential predictors of fracture. Logistic regression was used to generate predictors of fracture initially from FN-BMD. Next age, Cstress (as the most discriminating HSA-derived parameter), HAL and BMI were added to the model as potentially independent predictors. It was not necessary to include both HAL and Cstress in the logistic models, so the entire data set was examined without excluding the subjects missing HAL measurements. Cstress combined with age and BMI provided significantly better prediction of fracture than FN-BMD used alone as is current practice, judged by comparing areas under receiver operating characteristic (ROC) curves (p<0.001, deLong's test). At a specificity of 80%, sensitivity in identification was improved from 66% to 81%. Identifying women at high risk of hip fracture is thus likely to be substantially enhanced by combining bone density with age, simple anthropometry and data on the structural geometry of the hip. HSA might prove to be a valuable enhancement of DXA densitometry in clinical practice and its use could justify a more proactive approach to identifying women at high risk of hip fracture in the community. PMID- 11883409 TI - Prevalence of reduced bone mineral density in patients with anti-neutrophil cytoplasmic antibody associated vasculitis and the role of immunosuppressive therapy: a cross-sectional study. AB - Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis is a relapsing remitting disease, which is treated with corticosteroids (CS) in combination with cyclophosphamide. One of the major side-effects of this treatment is osteoporosis, which may result in the increased occurrence of fractures. In the present study we measured the prevalence of reduced bone mineral density (BMD) in a cross-sectional cohort of patients and correlated BMD findings with cumulative doses of CS and/or cyclophosphamide. BMD was measured by dual-energy X-ray absorptiometry (DXA) of the lumbar spine, radius and proximal femur between January 1998 and December 1999. Cumulative doses of CS and cyclophosphamide were calculated by chart review. Ninety-nine consecutive patients (48 men, 51 women) aged 55 +/- 16 years (mean +/- SD) were studied 50 months (median; range 0-400 months) after a diagnosis of ANCA-associated vasculitis had been made. Sixty-nine patients were treated with 10.7 g (median cumulative dose; range 0.4-67.2 g) of CS, and 88 patients were treated with 34.1 g (median cumulative dose; range 0.8 324.3 g) of cyclophosphamide. Fifty-seven percent of the patients had osteopenia (T-score: -1 to -2.5 SD), and 21% had osteoporosis (T-score: <-2.5 SD) at least at one site. Thirty-four of 37 (92%) postmenopausal women, 9 of 14 (64%) premenopausal women, and 34 of 48 (71%) men had either osteopenia or osteoporosis. The mean age- and sex-adjusted BMD (Z-score) of the proximal femur in men was found to be significantly lower than zero. Cumulative dose of CS therapy showed an inverse relation with Z-scores at the lumbar spine (p = 0.035) and proximal femur (p = 0.011). Cumulative dose of cyclophosphamide was not correlated with Z-scores. Osteopenia and osteoporosis are thus frequently observed in patients with ANCA-associated vasculities. However, only in men is the mean Z-score significantly lower than zero. Cumulative dose of CS therapy is significantly associated with bone loss at the spine and femur. PMID- 11883410 TI - Relationships between intestinal calcium absorption, serum vitamin D metabolites and smoking in postmenopausal women. AB - Smoking has been associated with low bone density, fractures and poor intestinal calcium absorption. Calcium absorption is a critical factor in calcium balance in postmenopausal women but the mechanisms causing decreased absorption efficiency in postmenopausal smokers are controversial and poorly defined. We performed a cross-sectional study of 405 postmenopausal women attending a clinic for the management of osteoporosis to compare intestinal calcium absorption efficiency, serum vitamin D metabolites and parathyroid hormone levels in postmenopausal women who had never smoked, who were smokers previously or who were current smokers, to examine the relationships between these variables in smokers. Two hundred and fifty-two of the women had never smoked, 79 had smoked previously and 74 were current smokers. The hourly fractional rate of calcium absorption was similar in non-smokers and those who had previously smoked. Radiocalcium absorption was less in the 74 smokers compared with the 331 non-smokers [0.60 (0.29 SD) vs 0.71 (0.27); p = 0.004], as were serum calcitriol (p<0.001) and parathyroid hormone (PTH) (p<0.01). There was no difference in the relationship between calcium absorption and serum calcitriol between smokers (r = 0.38) and non-smokers (r = 0.28); hence the impaired calcium absorption in the smokers was almost entirely attributable to suppression of the PTH-calcitriol endocrine axis. In postmenopausal women smoking is associated with a reduction in calcium absorption efficiency due to suppression of the PTH-calcitriol axis. This impairment of calcium absorption could lead to accelerated bone loss and limit the usefulness of dietary calcium supplementation. PMID- 11883411 TI - Evaluation of a hip fracture risk score for assessing elderly women: the Melton Osteoporotic Fracture (MOF) study. AB - Risk assessment for osteoporotic fracture within a primary care context, in old age, has received little attention. We aimed to develop such a risk score and assess its feasibility and validity. This was a 100% population-based, prospective cohort study, with a minimum 5 1/2 year follow-up among women aged 70 years and over, set in a large single general practice in Melton Mowbray, Leicestershire, UK. The main outcome measures were hip fracture, death and migration. Baseline measures included calcaneal broadband ultrasound attenuation (BUA), reported falls, balance, previous fracture history, medical problems, visual acuity, foot problems, body size, lifestyle factors and cognitive impairment. Seventy percent of the sample (1289) participated, including those in residential accommodation. Independent predictors of hip fracture over 3 years were low weight, kyphosis, poor circulation in the foot, epilepsy, short-term use of steroids and poor trunk maneuver. Using the highest tertile, a risk score based on these variables identified 84% (95% CI: 70% to 98%) of the hip fractures with a specificity of 68% (95% CI: 65% to 71%). BUA did not independently predict hip fracture in women of this age group. This study shows that a combination of readily obtained risk factors can identify elderly women who will sustain a hip fracture in the next 3 years more accurately than bone measurements alone in younger women. It also suggests that a risk score approach to universal assessment in the elderly is a feasible proposition in the primary care setting. PMID- 11883412 TI - The potential for estradiol and ethinylestradiol degradation in English rivers. AB - Water samples were collected in spring, summer, and winter from English rivers in urban/industrial (River Aire and River Calder, Yorkshire, UK) and rural environments (River Thames, Oxfordshire, UK) to study the biodegradation potential of the key steroid estrogen 17beta-estradiol (E2) and its synthetic derivate ethinylestradiol (EE2). Microorganisms in the river water samples were capable of transforming E2 to estrone (E1) with half-lives of 0.2 to 9 d when incubated at 20 degrees C. The E1 was then further degraded at similar rates. The most rapid biodegradation rates were associated with the downstream summer samples of the River Aire and River Calder. E2 degradation rates were similar for spiking concentrations throughout the range of 20 ng/L to 500 microg/L. Microbial cleavage of the steroid ring system was demonstrated by release of radiolabeled CO2 from the aromatic ring of E2 (position 4). When E2 was degraded, the loss of estrogenicity, measured by the yeast estrogen screen (YES) assay, closely followed the loss of the parent molecule. Thus, apart from the transient formation of E1, the degradation of E2 does not form other significantly estrogenic intermediates. The E2 could also be degraded when incubated with anaerobic bed sediments. Compared to E2, EE2 was much more resistant to biodegradation, but both E2 and EE2 were susceptible to photodegradation, with half-lives in the order of 10 d under ideal conditions. PMID- 11883413 TI - Health of lake whitefish (Coregonus clupeaformis) with elevated tissue levels of environmental contaminants. AB - Lake whitefish (Coregonus clupeaformis) were collected in 1996 from the St. Lawrence River, Quebec, Canada. Histologic examination was performed on major organs of 497 specimens and on the liver of 48 additional individuals. Preneoplastic and neoplastic hepatic changes consisted of vacuolated cell (n = 65), clear cell (n = 17), and acidophilic (n = 16) foci of altered hepatocyte, hepatocellular carcinoma (n = 12), cholangioma (n = 5), and cholangiocarcinoma (n = 28). Six fish were intersexes (1.2%), and 11.7% of the ovaries (26/223) had ducts containing spermatogonia or more differentiated cells of the male germ cell line. Asynchronous nodular maturation of the testes was present in 8.2% of the male fish (22/267). The mean hepatic concentrations of various contaminants, including polychlorinated biphenyls (PCBs), chlorobenzenes, pesticides, and trace metals, were 6 to 8 times higher in lake whitefish than in three other fish species (Ictalurus punctatus, Catostomus commersoni, and Stizostedion vitreum) collected at the same site. Condition factor of lake whitefish from this study was lower than that previously reported 40 to 50 years ago at this site and from contemporary pristine sites in the Great Lakes, USA. The presence of liver neoplasms, gonadal lesions, and a decreased condition factor in lake whitefish from the St. Lawrence River may be etiologically related to elevated tissue concentrations of toxic chemical contaminants. PMID- 11883414 TI - An in situ bioassay for estuarine environments using the microalga Phaeodactylum tricornutum. AB - This study aimed at evaluating the potential of an in situ algal bioassay for routine toxicity estimates of potentially contaminated estuarine environments using the microalga Phaeodactylum tricornutum immobilized in alginate beads. The influence of the initial cell density in the beads and of salinity on algal growth was first investigated. The potential of the proposed bioassay was evaluated by comparing laboratory with in situ results. A good growth performance of P. tricornutum was observed at all starting densities of beads. Although the growth rate of P. tricornutum was significantly affected by salinity, acceptability criteria currently adopted in algal assays were met, indicating the suitability of P. tricornutum as a test species for bioassays in estuaries. The large differences observed between the laboratory and in situ responses of P. tricornutum were most probably due to the temperature and light conditions less favorable for algal growth in the field and to the lack of representativeness of water samples compared to the field fluctuating conditions. These results showed the need for in situ assessments, especially in estuarine environments influenced by tides. To a lesser extent, the bioassay itself may also have been responsible for the laboratory and field differences. Further improvements in the bioassay chambers and procedures were also discussed. PMID- 11883415 TI - Spatial trends and bioaccumulation of organochlorine pollutants in marine zooplankton from the Alaskan and Canadian Arctic. AB - Planktonic copepods (Calanus glacialis and C. hyperboreus; n = 37) and water (n = 19) were collected to examine the spatial distribution and bioaccumulation of organochlorine contaminants (OCs) in the Alaskan and Canadian Arctic. The rank order of total OC (sigma OC) group concentrations in Calanus samples was toxaphene > or = sigma polychlorinated biphenyls (PCBs) > sigma hexachlorcyclohexane (HCH) > sigma DDT > sigma chlordane-related compounds (CHLOR) > sigma chlorobenzenes (ClBz). The dominant analyte was alpha-HCH in all water and zooplankton samples. The most abundant toxaphene congener in water and zooplankton samples was the hexachlorobornane B6-923. Organochlorine contaminant group concentrations in Alaskan zooplankton and water samples were lower than those in samples collected from sites in the eastern Canadian Arctic. Comparison of PCB and toxaphene congener profiles in zooplankton and water samples suggests that biotransformation by cytochrome P-4502B isozymes is low in Calanus, and limited phase I metabolism may occur. The log relationship of bioaccumulation factor (log BAF) versus octanol-water partition coefficient (log Kow) relationship was near 1:1 for OCs within the log Kow range of 3 to 6. A curvilinear model provided a better relationship between these two variables when OC compounds with log Kow > 6 were included. These results suggest that hydrophobic OCs (log Kow 3-6) in Calanus species are at equilibrium with the water concentrations and that physical partitioning, rather than biotransformation, is the major factor governing OC profiles in marine zooplankton. PMID- 11883416 TI - Responses of biofilms to combined nutrient and metal exposure. AB - Numerous studies have reported marked sensitivities of diatom species to phosphate and organic pollution but have ignored interactions with other common contaminants. The aim of the present study was to investigate the single and joint effects of increased phosphate and metal (cadmium, zinc) concentrations on benthic diatom communities. Microalgal biofilms from a relatively unpolluted stream were exposed in the laboratory to Zn, Cd, and P, separately and in combination, in concentrations found at a polluted stream in the same catchment. The Zn concentration reduced algal growth in biofilms more than the Cd concentration. Phosphate compensated for the single effect of each metal but not for their combined effects. Diatom community changes were evaluated using water quality indices based on the empirical sensitivities of taxa to nutrients (TDI) and organic pollution (%PTV). Phosphate exposure resulted in an increase of the eutrophy rank and presumed pollution-tolerant taxa. In contrast, exposure to Zn, Zn+Cd, and Zn+Cd+P caused a marked reduction of the TDI and %PTV community values. The successional trends in the laboratory matched the observed differences in microphyte communities in the reference and polluted river stations. However, the autoecology of the species present also revealed that the resulting composition of diatom communities cannot be attributed solely to the direct toxic effects of metal and nutrients and their interaction. Observed changes in the relative abundance of species are also determined by their growth form and microdistribution in biofilms. PMID- 11883417 TI - Chlordane enantiomers and temporal trends of chlordane isomers in arctic air. AB - A 14-year data set (1984-1998) for chlordane compounds in arctic airwas examined to discern temporal trends. trans-Chlordane (TC), cis-chlordane (CC), and trans nonachlor (TN) declined significantly (p < 0.001-0.02), with apparent times for 50% reduction of 4.9-9.7 y. The isomer fraction of TC = (TC/(TC + CC) also declined significantly (p < 0.001 -0.014) over the same time period. The enantiomeric composition of TC and CC was determined in air samples collected at arctic stations in Canada (1993-1996), Russia (1994), and Finland (1998), and a temperate station on the Swedish west coast (1998). Enantiomer fractions, EF = (+)/[(+) + (-)], were significantly different from measured EFs of racemic standards (0.498-0.501) at all stations for TC (p < 0.001) and two stations for CC (p < 0.001 to <0.05). These observations suggest changing source composition of chlordane in arctic air, with a greater proportion of weathered residues in recent years, possibly derived from soils. Identification of nonracemic (mean EFs = 0.662-0.703) heptachlor exo-epoxide (HEPX) at the four air stations further exemplifies contributions of soil emissions to long-range transport of chlordane related compounds. PMID- 11883418 TI - Monitoring perfluorinated surfactants in biota and surface water samples following an accidental release of fire-fighting foam into Etobicoke Creek. AB - Perfluorinated surfactants have emerged as priority environmental contaminants due to recent reports of their detection in environmental and biological matrices as well as concerns regarding their persistence and toxicity. In June 2000, 22000 L of fire retardant foam containing perfluorinated surfactants was accidentally released at L. B. Pearson International Airport, Toronto, ON, and subsequently entered into Etobicoke Creek, a tributary to Lake Ontario. A suite of analytical tools that include liquid chromatography/tandem mass spectrometry (LC/MS/MS) and 19F NMR were employed to characterize fish (common shiner, Notropus cornutus) and surface water samples collected following the discharge of the perfluorinated material. Total perfluoroalkanesulfonate (4, 6, and 8 carbons) concentrations in fish liver samples ranged from 2.00 to 72.9 microg/g, and total perfluorocarboxylate (5-14 carbons) concentrations ranged from 0.07 to 1.02 microg/g. In addition to fish samples, total perfluoroalkanesulfonate (6 and 8 carbons) concentrations were detected in creek water samples by LC/MS/MS over a 153 day sampling period with concentrations ranging from <0.017 to 2260 microg/L; perfluorooctanoate concentrations (<0.009-11.3 microg/L) were lower than those observed for the perfluoroalkane-sulfonates. By 19F NMR, the total perfluorinated surfactant concentrations in surface water samples ranged from < 10 to 17000 microg/L. A bioaccumulation factor range of 6300-125000 was calculated for perfluorooctanesulfonate, based on concentrations in fish liver and surface water. The residence time of perfluorooctanesulfonate in Etobicoke Creek as well as the high bioaccumulation in fish liver suggests that perfluorinated surfactants will persist and bioaccumulate following release into the aquatic environment. PMID- 11883419 TI - Measurement of emissions from air pollution sources. 4. C1-C27 organic compounds from cooking with seed oils. AB - The emission rates of gas-phase, semivolatile, and particle-phase organic compounds ranging in carbon number from C1 to C27 were measured from institutional-scale food cooking operations that employ seed oils. Two cooking methods and three types of seed oils were examined: vegetables stir-fried in soybean oil, vegetables stir-fried in canola oil, and potatoes deep fried in hydrogenated soybean oil. The emission rates of 99 organic compounds were quantified, and these include n-alkanes, branched alkanes, alkenes, n-alkanoic acids, n-alkenoic acids, carbonyls, aromatics, polycyclic aromatic hydrocarbons (PAH), and lactones. Carbonyls and fatty acids (n-alkanoic and n-alkenoic acids) make up a significant portion of the organic compounds emitted from all three seed oil cooking procedures. The compositional differences in the organic compound emissions between the different cooking operations are consistent with the differences in the organic composition of the various cooking oils used. The distribution of the n-alkanoic acids between the gas and particle phases was found to be in good agreement with gas/particle partitioning theory. The relative importance of emissions from commercial deep frying operations to the total emissions of C16 and C18 n-alkanoic acids in the Los Angeles urban area was estimated using the available information and is estimated to account for approximately 7% of the total primary emissions of these acids. Additional emissions of these n-alkanoic acids from stir-frying and grill frying operations are expected. Estimates also indicate that seed oil cooking may make up a significant fraction of the emissions of lighter n-alkanoic acids such as nonanoic acid. PMID- 11883420 TI - Influence of phosphate on bacterial adhesion onto iron oxyhydroxide in drinking water. AB - The transport and storage of drinking water in water distribution systems can modify its initial composition and properties. The accumulation of bacteria on corroded pipes is prejudicial and may lower the microbiological quality of the water. Previous results have shown that when pipes are highly corroded, the addition of phosphate, used as an anticorrosion treatment, decreases the bacterial concentration in the water. We studied the possibility of using phosphate to reverse the surface charge of iron oxyhydroxide (FeOOH) to limit bacterial adhesion. Iron oxyhydroxide (IOH) particles and Escherichia coli SH 702 were used as models of corrosion products and bacterial contamination, respectively. Electrophoresis was used to characterize the initial surface charges of both types of particles and the modifications that occurred after the addition of phosphate anions. Flow cytometry and adhesion assays were used to build adsorption isotherms of bacteria on IOH versus (phosphated-) IOH. X-ray photoelectron spectroscopy permitted to determine the chemical composition of the E. coli envelope and to discuss on functional groups responsible for bacterial surface properties. In the present conditions, adding phosphate to water allowed a decrease of 75% of the bacteria adhering to IOH. PMID- 11883421 TI - Effect of selected literature on dentists' decisions to remove asymptomatic, impacted lower third molars. AB - The aim of this study was to assess the effect of studying selected literature on dentists' decisions to remove asymptomatic, impacted lower third molars. A pre test-post-test control group design was used. Given 36 patient cases, two groups of 16 general dental practitioners each were asked to assess the need for removal of asymptomatic impacted lower third molars. The cases were classified by three parameters: 'position of the third molar', 'impaction type', and 'patient age'. After studying selected literature on this subject by the intervention group, both groups were asked to assess the same cases again. Frequencies of decisions to remove the third molars were calculated. For each participant, tables were composed by crosstabulating the indication to remove a third molar with each of the three parameters. T-tests were used to test the significance of the difference between pre-test and post-test decisions. The overall number of indications to remove asymptomatic, impacted lower third molars decreased by 37% in the intervention group. In the control group, the difference between pre- and post-test was not statistically significant. It was concluded that the provision of selected literature significantly influences treatment decision making by dentists in a third molar decision task. PMID- 11883422 TI - In vivo wear of three types of veneering materials using implant-supported restorations: a method evaluation. AB - In a previous study, we determined the precision and the positional duplicability of a system that used implant connectors to transfer restorations between the oral cavity and the measuring device. The objectives of the present study were to determine whether this testing procedure was suitable for clinical use, and to apply the procedure to the longitudinal assessment of the wear of three types of veneering materials. Ten patients received a total of 12 restorations. The restorations were made either of ceramic (positive control), poly(methylmethacrylate) resin (negative control) or of composite resin. The restorations were profiled at baseline and at 3, 6, 9, 12, 18 and 24 months. For profiling, the restorations were secured to the x-y table of the measuring device using the octagonal connector of the ITI implant system. Numerical analyzes were performed using a commercial array-oriented software package. The ceramic and the composite wore at roughly 12-14 microm yr(-1) (height) 10-12 microm3 microm2 yr( 1) (volume). The resin wore at c. 50 micorm yr(-1) (height) and c. 45 microm3 microm2 yr(-1) (volume). The error of the procedure was estimated at +/- 13%. It was concluded that the procedure was applicable for clinical studies, and that the composite did not differ from the ceramic as to its wear rate. PMID- 11883423 TI - Straelensiosis in dogs: a newly described nodular dermatitis induced by Straelensia cynotis. AB - A distinctive nodular dermatitis induced by Straelensia cynotis, a newly described trombidioid larval mite which resides in hair follicles, was identified in 12 dogs living in France. They all had scattered, small (1 to 3 mm in diameter), pale, firm skin nodules, variable in distribution but always affecting the dorsal regions of the head and trunk; they were distributed over the whole body of seven of the dogs. The animals were otherwise healthy except for three severely infested fox terriers which had a decreased appetite, were lethargic, and whose skin nodules were painful to the touch. The nodules did not induce pruritus. The lesions usually began as erythematous papules which developed into firm pale nodules. The dermatitis resolved within two to 12 months. Topical acaricides were ineffective but the skin nodules regressed after treatments with systemic avermectins. Histologically, each nodule was composed of a dilated follicular ostium containing a well-preserved larval mite, and showed a pseudoepitheliomatous follicular hyperplasia and an abundant perifollicular mucinosis. The larvae were identified as belonging to the genus Straelensia (Acari: Leeuwenhoekiidae). It was clearly established that the three fox terriers had become infested within a fox's den. The nymphs and adults of this species of mite are believed to live in foxes' dens; foxes are considered to be the natural host for the larval stage, and dogs a permissive but occasional host. PMID- 11883424 TI - Vets and the future of farming. PMID- 11883426 TI - Judging survey quality: local variances. AB - The present study was undertaken to explore possibilities to judge survey quality on basis of a limited and restricted number of a-priori observations. Here, quality is defined as the ratio between survey and local variance (signal-to noise ratio). The results indicate that the presented surveys do not permit such judgement. Furthermore, the discussion also suggests that the 5-fold local sampling strategies do not merit any sound judgement in about 10% of all cases. The results further imply that surveys will benefit from procedures, controls and approaches in sampling and sample handling, to assess both average, variance and the nature of the distribution of elemental concentrations in local sites. This reasoning is compatible with the idea of the site as a basic homogeneous survey unit, which is implicitly and conceptually underlying any survey performed. PMID- 11883425 TI - Identification of a novel family of sequence repeats among prokaryotes. AB - The rapid increase in genomic sequences provides new opportunities for comparative genomics. In this report, we describe a novel family of repeat sequences that is present in Bacteria and Archaea but not in Eukarya. The repeat loci typically consisted of repetitive stretches of nucleotides with a length of 25 to 37 bp alternated by nonrepetitive DNA spacers of approximately equal size as the repeats. The nucleotide sequences and the size of the repeats were highly conserved within a species, but between species the sequences showed no similarity. Due to their characteristic structure, we have designated this family of repeat loci as SPacers Interspersed Direct Repeats (SPIDR). The SPIDR loci were identified in more than forty different prokaryotic species. Individual species such as Mycobacterium tuberculosis contain one SPIDR locus, while other species such as Methanococcus jannaschii contained up to 20 different loci. The number of repeats in a locus varies greatly from two repeats to several dozens of repeats. The SPIDR loci were flanked by a common 300-500-bp leader sequence, which appeared to be conserved within a species but not between species. The SPIDR locus of M. tuberculosis is extensively used for strain typing. The finding of SPIDR loci in other prokaryotes, including the pathogens Salmonella, Campylobacter, and Pasteurella may extend this surveillance to other species. PMID- 11883427 TI - Repetitive motion hand disorders. AB - The clinical management of cumulative trauma disorder is based upon the identification and treatment of individual component pathologies and, frequently, referral to a knowledgeable occupational therapist with an understanding of ergonomic behavioral, postural, and workspace modification. Most commonly these individual pathologic entities are carpal tunnel syndrome, cubital tunnel syndrome, trigger finger, and De Quervain's tenosynovitis. In this article, the anatomy, diagnosis, and treatment of each of these disorders will be considered separately. In addition, since these clinical entities are often use-related, special attention should be directed toward biomechanical and ergonomic considerations. PMID- 11883428 TI - Using genetic markers for disease resistance to improve production under constant infection pressure. AB - Animals will show reduced production when exposed to a constant infection pressure unless they are fully resistant, the size of the reduction depending on the degree of resistance and the severity of infection. In this article, the use of QTL for disease resistance for improving productivity under constant infection pressure is investigated using stochastic simulation. A previously published model was used with two thresholds for resistance: a threshold below which production is not possible and a threshold above which production is not affected by the infection. Between thresholds, observed production under constant infection is a multiplicative function of underlying potential production and level of resistance. Some simplifications of reality were adopted in the model, such as no genetic correlation between potential production and resistance, the absence of influence of lack of resistance on reproductive capacity, and the availability of phenotypes in both sexes. Marker-assisted selection was incorporated by assuming a proportion of the genetic variance to be explained by the QTL, which thus is defined as a continuous trait. Phenotypes were available for production, not for resistance. The infection pressure may vary across time. Results were compared to mass selection on production under constant as well as intermittent infection pressure, where the infection pressure varied between but not within years. Selection started in a population with a very poor level of resistance. Incorporation of QTL information is valuable (i.e., the increase in observed production relative to mass selection) when a large proportion of the additive genetic variance is explained by the QTL (50% genetic variance explained) and when the heritability for resistance is low (h2R = 0.1). Under constant infection pressure, incorporating QTL information does not increase selection responses in observed production when the QTL effect explains less than 25% of the genetic variance. Under intermittent selection pressure, the use of QTL information gives a slightly greater increase in observed production in early generations, relative to mass selection on observed production, but still only when the QTL effect is large or the heritability for resistance is low. The additional advantage of incorporating QTL information is that use of (preventive) medical treatment is possible, or animals may be evaluated in uninfected environments. PMID- 11883429 TI - Growth performance and whole-body composition of pigs experimentally infected with Mycoplasma hyopneumoniae. AB - Mycoplasma hyopneumoniae (Mh) is the primary infectious pathogen responsible for enzootic pneumonia in pigs. Although Mh is thought to impair growth performance, whole-body composition, and fat and protein accretion in pigs with pneumonia have not been reported and the mechanism through which Mh reduces growth is unknown. The objectives of this study were to evaluate the effects of Mh on growth performance, whole-body composition, and protein and fat accretion in nursery pigs and to determine whether Mh infection increases the expression of interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha). Sixty four 2-wk-old Mh-free pigs were used (two trials) in a randomized complete block design. In each trial, two pigs were housed in each of 16 disease-containment chambers. At 4 wk of age, pigs were inoculated intratracheally with 3 mL of Mh broth (P5722-3, 10(7) cfu/mL) or sterile Friis culture medium. Clinical signs of disease and feed intake were monitored daily and body weight was determined weekly for 4 wk. Whole-body composition was determined from pigs killed 0, 14, and 28 d after inoculation, and the comparative slaughter technique was used to estimate protein and fat accretion. At death, gross lung lesions were quantified, and lung tissue was collected to verify the presence or absence of Mh, and to determine cytokine mRNA levels. Control pigs displayed no overt signs of infection and were Mh-negative and free of pulmonary lesions. Pigs inoculated with Mh showed pneumonic coughing (P < 0.005), were Mh-positive, and had pulmonary lesions that affected 4.5% (P < 0.01) and 14.1% (P < 0.001) of total lung surface area at 14 and 28 d, respectively, after inoculation. Ribonuclease protection assays revealed increased IL-1beta (P < 0.04) and TNF-alpha (P < 0.06) mRNA in lung tissue collected from a lesion site compared with tissue collected 10 cm from a lesion site or from control pigs. Interestingly, Mh did not depress weight gain or feed efficiency during any week of the 28-d study (P > 0.10). Moreover, Mh did not affect whole-body fat or protein accretion (P > 0.10). Thus, in spite of inducing disease and expression of inflammatory cytokines, Mh alone did not affect growth performance and whole-body composition of nursery pigs during the 4-wk experiment. The ability of pigs to contend with Mh may have resulted from the absence of other pathogens that generally co-exist with Mh under commercial conditions. PMID- 11883430 TI - Evaluation of mobile nylon bag technique for determining apparent ileal digestibilities of protein and amino acids in growing pigs. AB - The mobile nylon bag technique (MNBT) may offer a simple, rapid means for assessing ileal AA digestibility of pig feed ingredients. In the present study, the effects of washing bags recovered from digesta, the amount and fineness of feeds, and feed trypsin inhibitor activity on apparent ileal digestibilities (AID) of CP and AA were determined with the MNBT. Twenty-four ileorectal anastomosed pigs (Yorkshire x Chinese Black barrows, 30 kg initial BW), of which 12 were fitted with duodenal T-cannulas, were used. Not washing the bags recovered from ileal digesta resulted in a reduction (P < 0.05) in apparent ileal digestibilities of CP and AA determined by MNBT. Washing the bags for more than 4 min overestimated (P < 0.05) the apparent ileal digestibilities of CP and AA compared with those determined with the anastomosed pigs. Sample size and fineness of grinding also affected (P < 0.05) apparent ileal digestibilities of CP determined by MNBT. The apparent ileal digestibilities of CP determined by MNBT were reduced (P < 0.05) when sample size exceeded 0.75 g and when feed was ground through screens with a mesh size of more than 1.0 mm. The closest agreement between results obtained by MNBT and a conventional ileal digestibility assay occurred when 0.75 g of feed ground through a 1.0-mm mesh screen was used per bag and bags were washed for 2 min after retrieval from digesta. Further studies are warranted to investigate the use of the mobile nylon bag technique for predicting the ileal digestibilty of AA for feeds containing antinutritional factors. PMID- 11883432 TI - Evaluation of the homoarginine technique for measuring true ileal amino acid digestibilities in pigs fed a barley-canola meal-based diet. AB - The homoarginine technique has been suggested as a means to determine true ileal amino acid digestibilities in nonruminant animals fed protein-containing diets. Conditions for guanidinating lysine to homoarginine in barley and canola meal and the effect of this process on nutrient composition and ileal digestibilities in the resulting material were investigated. Conditions tested were methylisourea concentration (0.4, 0.5, or 0.6 M) and reaction time (4 or 6 d) at pH 10.5. Using 0.4 methylisourea M solution for 4 or 6 d gave guanidination rates of 72.5 and 78.5% for barley and 72.3 and 75.2% for canola meal, respectively. Using 0.5 M gave 88.0 and 84.6% guanidination rates in barley and canola meal, respectively, after a 6-d reaction time. Under these conditions, guanidination did not change the nutrient composition of barley (P > 0.10), whereas it increased CP (38.4 vs 49.0%), crude fiber (10.2 vs 16.0%), acid detergent fiber (30.0 vs 43.4%) and neutral detergent fiber (29.8 vs 49.4%) levels in canola meal (P < 0.05). Four 33.6-kg barrows fitted with a simple T-cannula at the terminal ileum were fed a 16% CP unguanidinated barley and canola meal-based diet for four consecutive 14-d periods. Ileal digesta were collected continuously for 24 h on d 12 and 14 to determine apparent nutrient digestibilities. On the morning of d 14, pigs were fed a diet in which half of the barley and canola meal was replaced with guanidinated material for determining true ileal amino acid digestibilities. Digesta samples were pooled by pig and by 24-h period to give 16 observations per diet. Apparent ileal digestibilities of DM, CP, and AA in the unguanidinated and guanidinated barley-canola meal diet were similar (P > 0.10) despite the changes observed in canola meal. Apparent ileal lysine digestibility was 73.9 and 74.5% in the unguanidinated and guanidinated diet, respectively. The true ileal lysine digestibility was 88.1%. The present results show that guanidination does not interfere with digestion and further support the use of the homoarginine method for determining true ileal amino acid digestibilities in pigs fed practical diets. A methylisourea solution of 0.5 M and a 6-d reaction time are recommended for converting lysine to homoarginine in barley and canola meal. PMID- 11883431 TI - Regional and processor variation in the ileal digestible amino acid content of soybean meals measured in growing swine. AB - To assess differences in soybean meal quality related to region of production, researchers in Illinois, Kansas, North Carolina, The Netherlands, and Ohio collected four soybean meal samples processed locally at least 15 d apart. These samples were assayed for ileal amino acid digestibility by pigs using a common soybean meal and a soy protein concentrate as references, and a low-protein casein diet for determination of endogenous amino acid losses. Digestibility was determined at each university using seven barrows surgically fitted with ileal cannulas in a 7 x 7 Latin square design. The experimental diets contained 17% CP from the test material except for the low-protein casein diet. Animals were fed twice daily, 12 h apart, at a level of 45 g x kg(-0.75) BW for each meal. Following a 5-d adaptation period, ileal digesta were collected for two 12-h periods for 2 d to be used for determination of ileal digestibility. Variation in amino acid digestibility was very small among and within sites and was much smaller than variation in the concentration of amino acids. Among sites, samples from The Netherlands had less total and thus digestible lysine and methionine than the U.S. samples (P < 0.05). The soybean meals tested in this experiment were approximately 4% higher in amino acids than that reported in the NRC (1998). True (standardized) digestibilities, however, were very similar to NRC values except for cysteine and threonine, which were 5 and 3 percentage points lower in this experiment, respectively. In conclusion, soybeans grown in the United States and locally processed into soybean meal were very similar in nutritional composition. Soybean meals produced in The Netherlands were lower in lysine and methionine (P < 0.05) but had a digestibility similar to that produced in the United States. PMID- 11883433 TI - Effect of chromium tripicolinate supplementation on porcine immune response during the postweaning period. AB - A total of 60 pigs from 15 litters were used during two experiments to assess the effects of maternal supplementation of Cr tripicolinate on performance and immune status of the offspring during the first 42 d after weaning. Gilts were raised on diets with either 0 (-Cr) or 200 (+Cr) ppb supplemental Cr from Cr tripicolinate. Their offspring were weaned at about 23 d of age. Pigs from dams fed supplemental Cr were fed Cr-supplemented diets after weaning and pigs from unsupplemented dams were not fed supplemental Cr. Pigs were housed in groups of three according to litter origin (six -Cr and nine +Cr) and BW. Feed and water were available for ad libitum consumption. Serum was collected 24 h after birth, the day after weaning (d 0), and, subsequently, every 7 d through d 28 for measurement of total immunoglobulin (Ig) G and IgM. For the 42-d nursery period, ADG (511 and 531 g/d for -Cr and +Cr, respectively), ADFI (827 and 851 g/d), and gain:feed (0.62 and 0.62) were not affected (P > 0.15) by Cr. Total IgG was not affected (P > 0.10) by Cr on d 0 (7.6 and 7.7 mg/mL for -Cr and +Cr, respectively) or on d 7 (6.3 and 6.1 mg/mL for -Cr and +Cr, respectively), when the lowest concentrations were observed. By d 28, total IgG had risen (11.7 and 8.9 mg/mL for -Cr and +Cr, respectively) and was affected by Cr (P = 0.03). Total IgM followed a similar pattern and was lowest on d 7. In each trial, IgG at 24 h after birth was inversely related to IgG from d 14 through 28 in the nursery. Additionally, the lowest Ig concentrations occurred between 4 and 5 wk of age, rather than at weaning (23 d of age); this may represent a vulnerable period for weanling pigs. Supplementation of the weanling pig diets with Cr tripicolinate did not significantly affect performance and immune status during the postweaning period. PMID- 11883434 TI - Effect of chromium tripicolinate supplementation on porcine immune response during the periparturient and neonatal period. AB - A total of 36 gilts were used to assess the effects of Cr tripicolinate supplementation on immune response in sows and their offspring during the periparturient and neonatal period. Gilts were raised from weaning to reproductive age on diets with either 0 (-Cr) or 200 (+Cr) ppb supplemental Cr from CrPic. Subsequently, 22 gilts (9 -Cr and 13 +Cr) in parity 1 and 16 sows in parity 2 (7 -Cr and 9 +Cr) underwent immune status testing. Only sows that completed all procedures in parity 1 were included in parity 2. Sows were immunized with ovalbumin about 3 wk (d 0), and again 14 d later for gilts, prior to anticipated farrowing, and serum was collected on d 0 and at 14-d intervals for a total of four samples. Serum was collected from five to six pigs/litter at 24 h after birth, three or six pigs/litter the day after weaning (25 d of age) in parity 1, and three pigs/litter the day of weaning (20 d of age) in parity 2. Milk was collected at 1 h (colostrum), 6.5 d (early), and 19 d (late) after farrowing. The only effect of Cr on total immunoglobulin (Ig) concentration was on sow serum IgG (21.7 and 24.1 mg/mL for -Cr and +Cr, respectively; P = 0.08) and IgM (11.0 and 12.5 mg/mL; P = 0.06) on d 0. No effect (P > 0.15) of Cr was observed on the IgG antibody response to ovalbumin, but Cr was associated (P < 0.10) with a decreased IgM antibody response to ovalbumin beginning on d 14. In parity 2, colostral total IgG increased (80.6 and 92.4 mg/mL for parity 1 and 2, respectively; P = 0.06), which was reflected in the neonates at 24 h after birth (33.6 and 39.7 mg/mL; P = 0.01) and at weaning (7.3 and 13.3 mg/mL; P < 0.001). Supplementation of Cr tripicolinate had minimal effects on humoral antibody response of the dam or its transfer to the neonate; however, parity greatly influenced the concentrations of immunoglobulins in the milk and their transfer to the neonate. PMID- 11883435 TI - Assessment of chromium tripicolinate supplementation and dietary energy level and source on growth, carcass, and blood criteria in growing pigs. AB - Two experiments were conducted to evaluate potential interactive effects of supplemental Cr and dietary energy supply in growing pigs. Experiment 1 used 36 individually penned barrows, 25 to 65 kg, in a 2 x 3 factorial arrangement of supplemental Cr (0 or 200 ppb) and energy level (70, 80, or 90% of ME requirement). A corn-soybean meal basal diet was designed to supply all protein, mineral, and vitamin needs and 70% of the estimated ME need at 70% of ad libitum feed intake. Additional energy to 80% or 90% of the ME requirement was provided by a cornstarch/corn oil blend. In Exp. 2, 30 individually penned barrows, 23 to 68 kg, were used in a 2 x 4 factorial arrangement of supplemental Cr (0 or 200 ppb) and added energy source (none, cornstarch, corn oil, or choice white grease) with basal diets identical to Exp. 1. The various energy sources were added to 90% of the ME requirement. In both experiments, growth data were collected over a 50-d period and pigs were killed at 70.1 kg. Increasing energy levels increased (linear, P < 0.01) ADG, average backfat thickness, 10th rib backfat thickness, and cooler shrink and decreased (linear, P < 0.01) longissimus muscle area in Exp. 1. Carcass composition increased (linear, P < 0.01) in lipid and decreased in protein, water, and protein:lipid ratio in response to increasing ME levels. Similar results were observed in Exp. 2 in response to added energy, regardless of the energy source used. In response to ME, linear increases (P < 0.05) in plasma insulin concentration before feeding and after feeding were observed in Exp. 1. In Exp. 2, plasma insulin concentration was lower for the basal diet before feeding (P < 0.05) and higher for the starch diet after feeding (P < 0.01); insulin:glucose ratio increased (P < 0.01) after feeding for starch compared to oil and fat. No consistent effect of Cr or Cr x ME level on performance or carcass was observed (P > 0.10) in these experiments. Similarly, no Cr effect or Cr x ME interaction (P > 0.10) was observed in plasma glucose or insulin levels. Dietary energy levels markedly affected growth criteria in growing pigs (23 to 68 kg) in these experiments, as anticipated, but supplemental Cr was without effect on performance or carcass responses. PMID- 11883436 TI - Genetic basis of familial Meniere's disease. AB - The genetic basis of familial Meniere's disease (MD) is unclear. We present a genetic investigation of six individuals in two families with familial MD. Linkage analysis was performed using polymorphic DNA markers linked to the human leukocyte antigen (HLA) locus that map to chromosome 6p. We have demonstrated the presence of anticipation in successive generations and the absence of HLA association. This is the second report of anticipation in familial MD in the literature, and it suggests that efforts should be directed toward finding a trinucleotide expansion as a possible genetic lesion in this uncommon disorder. PMID- 11883437 TI - Clinical decision analysis in the treatment of acute otitis media in a child over 2 years of age. AB - Acute otitis media (AOM) is the most common infection diagnosed in children. In Canada and the United States, the standard of care for treatment of children over 2 years of age diagnosed with AOM is a course of antibiotics for 5 to 10 days. However, in other countries, treatment is primarily symptomatic, and antibiotics are prescribed only if symptoms fail to resolve. Clinical decision analysis is a process whereby different treatment options are assessed systematically. All clinical pathways are incorporated into a model, probabilities for each event are determined from the literature, and clinical outcomes are quantified as to the preference of patients. The decision analysis then determines the most appropriate treatment option for the disease process. For AOM in a child over 2 years of age, four treatment options were considered including observation followed by 10 days of antibiotic therapy if required for failure of symptoms to resolve, observation followed by 5 days of antibiotic therapy if required, 10 days of antibiotic therapy when the child was initially diagnosed with AOM, and 5 days of antibiotic therapy when the child was initially diagnosed with AOM. Using a clinical decision analysis model for the treatment of AOM in a child over 2 years of age, the most appropriate treatment was found to be initial observation followed by 5 days of an antibiotic if the child failed to improve spontaneously. The decision analysis model developed was designed to be free of construction bias and was found to be robust in multiple sensitivity analysis. PMID- 11883438 TI - Cochlear implantation and Pendred's syndrome mutation in monozygotic twins with large vestibular aqueduct syndrome. PMID- 11883440 TI - Minor role for BRCA2 (exon11) and p53 (exon 5-9) among Sudanese breast cancer patients. AB - A cohort of 20 breast cancer patients from the Sudan was tested for germ line and somatic mutation in their BRCA2 exon 11 as well as the main conserved area of the p53 tumor suppressor gene. The results indicate that both regions may play a limited role in the pathogenesis of breast cancer in those patients. The fact that there are no somatic mutations detected in p53 was particularly surprising as the expected rate for mutations in breast cancer is 30-50%. PMID- 11883439 TI - Irofulven induces apoptosis in breast cancer cells regardless of caspase-3 status. AB - Caspase-3 deficiency can limit the efficiency of pro-apoptotic anticancer treatments. Irofulven (hydroxymethylacyl-fulvene, HMAF. MGI 114, NSC 683863) is an antitumor drug, currently in a Phase III and multiple Phase II trials, which can differentiate between tumor and normal cells in apoptosis induction. This study investigated whether apoptosis induced by irofulven requires caspase-3. Irofulven action was compared in breast cancer cells differing in caspase-3 status: deficient MCF-7 cells and proficient MDA-MB-231 cells and in normal human mammary epithelial cells, HMEC. Irofulven induces significant, concentration and time-dependent apoptotic DNA fragmentation in breast cancer cell lines, regardless of caspase-3 status. After 12, 24 and 48 h incubation at 1 microM irofulven (approximately 3 x GI50), fragmented DNA comprised 3.7, 14.1 and 34.6% and 8.4, 12.6 and 20.3% of total DNA in MCF-7 and MDA-MB-231 cells, respectively. Cell viability (trypan blue exclusion) remained largely unaffected during the first 24 h but decreased markedly after 48 h, indicating secondary necrosis. Net losses in cell numbers were apparent at 48 h. Normal HMEC cells were refractory to 1 microM drug with only approximately 3-9% fragmented DNA after 12-48 h, although apoptosis was observed at drug levels >3 microM. The broad-spectrum caspase inhibitor Z-VAD-fmk inhibited irofulven-induced apoptosis of all cell lines at 20 microM with nearly complete abrogation of apoptosis at 100 microM. Irofulven treatment resulted in marginal caspase-3 processing in MDA-MB-231 and HMEC cells. These results indicate that whereas the caspase cascade mediates irofulven- induced apoptosis, caspase-3 is dispensable (supported by NIH CA70091 and CA78706). PMID- 11883441 TI - [Vilnius Medical Association: the first half-century (1805-1850)]. AB - Vilnius Medical Association, oldest scientific medical association in Poland and in the Russian Empire, was founded for the purposes of self-development in 1805 on the initiative of an Austrian professor of pathology and detailed therapy, Jozef Frank (1771-1842). From its beginnings, the association acted for the good of society and science, independently of political conditions and with varying intensity. Apart from doctors, it also included pharmacists who contributed towards raising the standard of pharmacy in Lithuania. The first half-century of its existence constitutes the most fruitful period of activity as it was supported by professors of the then Medical Department of Vilnius University. At that time the Association established its regulations, founded the Library, and published "Pamietnik Wilenskiego Towarzystwa Lekarskiego" (Journal of Vilnius Medical Association, 1818 and 1821), "Pamietnik Farmaceutyczny" (Pharmaceutical Journal, 1820 and 1821), "Dziennik Medycyny Chirurgii i Farmacji" (Journal of Medicine, Surgery and Pharmacy, 1820, 1823, and 1830), as well as "Collectanea Medico Chirurgica", 1838. It also began gathering and analysing data for medical statistics, as well as developing Polish medical and chemical nomenclature. The Association re-organised shipments of medicines from pharmacies. It endeavoured to raise the standard of medical knowledge in the Vilnius area, combated epidemics of plague, cholera, scarlet fever, smallpox, typhus, diphteria, dysentery and monitored occurrences of plica. It also established the first Institute of Vaccination against Smallpox in Vilnius (1809); carried out scientific research on health resorts at Druskienniki, Birsztany and Stokliszki; promoted hygiene and examined the health of the inhabitants of Vilnius and the surrounding area as well as endorsed authority decisions related to the location of temporary military hospitals. At its meetings, members of the Association discussed their own medical cases as well as methods of treatment, presented the most recent reports in the medical literature, combated obsolete theories and scientific systems and established cooperation with other scientific associations in the country as well as abroad. World famous scientists were also members of the Vilnius Medical Association. PMID- 11883442 TI - Human reliability and confinement. AB - Problems inherent in the modifiability of circadian periodicity and in impoverished sensory environments were explored for the purpose of appraising attenuative effects upon human reliability. Accordingly, highly selected subjects were confined within a one-man altitude chamber for prolonged periods of time and under a variety of designed conditions. The findings relative to the modifiability of biological rhythm indicate that adjustment to a drastic revision of the 24-hour biological day was accomplished to a significant and practical extent by certain subjects, the extent of adjustment was directly related to the maintenance of high initial levels of proficiency, and just as subjects differ greatly in their adjustment to revised biological time, they differ to an equal extent in the degree of synchronization manifested by the apparent periodicities of the different physiological systems. In the investigation of impoverished sensory environments, it was found that the joint effects of impoverished sensory conditions and continuous work at an operator system drastically degraded the reliability of certain subjects. Further, neither prior experience nor knowledge acted to mitigate the degree of aberrancy experienced which in the case of one subject was so extreme as to necessitate his removal from the chamber prior to the termination of confinement period. Finally, management of certain aberrant behavior, specifically hallucinatory experiences, could be successfully achieved by those subjects who continuously attempted to maintain a diversity of sensory input. PMID- 11883443 TI - Biological contamination of Mars. I. Survival of terrestrial microorganisms in simulated Martian environments. AB - It has been postulated that the accidental introduction of terrestrial microorganisms to other planets during the course of space exploration might impede or bias the detection of organic matter and possible indigenous organisms, and thereby confuse subsequent studies of extraterrestrial life. To assess the likelihood of biological contamination of Mars, we have applied the principle of natural selection on a laboratory scale. Terrestrial microorganisms were collected from a variety of environments, including regions of high alkalinity, low mean daily temperature, and low annual rainfall. The air-dried soils were then subjected to a simulated Martian environment involving 12-hour freeze-thaw cycles from about -60 degrees C to about +20 degrees C; atmospheres of 95 per cent nitrogen, 5 percent carbon dioxide and low moisture content: < or = 0.1 atm pressure; and a total ultraviolet dose at 2537 angstrom of 10(9) erg cm-2. In some experiments, organic supplements were provided. Survivors were scored on supplemented agar. Preliminary results indicate a wide variety of survivors, even when no organic supplements were introduced. Survivors included obligate and facultative anaerobic spore-formers and non-spore-forming facultative anaerobic bacteria. Diurnal freezing and thawing was continued for six months. There was no significant loss of viability after the first freeze-thaw cycle. An extensive literature survey shows that survival of terrestrial microorganisms under individual simulated Martian conditions has been known for decades. The present investigation shows the absence of pronounced synergistic effects inhibiting survival. The probable existence of organic matter and moisture on Mars, at least in restricted locales and times, makes it especially likely that terrestrial microorganisms can also reproduce on Mars. The demonstration that all samples of terrestrial soil tested contain a population of microorganisms which survive in simulated Martian environments strongly underscores the need for scrupulous sterilization of all spacecraft intended for Mars landing. PMID- 11883444 TI - Some biological and physical factors in dry heat sterilization: a general review. AB - There is a surprising lack of quantitative data on sterilization by dry heat so that microbiologists have little knowledge of the role played by various biological and physical factors in this sterilizing process. A recent investigation by the author has shown that the aerobic mesophilic bacterial sporeformers, such as Bacillus subtilis and Bacillus coagulans, are the most resistant among several species of sporeforming bacteria to dry heat sterilization. The type of carrier on which the spores are exposed to dry heat markedly affects their thermal resistance. An analysis of four carriers showed that spores on sand or vermiculite are more difficult to destroy than spores on paper or glass. Spores under low vacuums are more susceptible to dry heat sterilization than spores in helium, which are more susceptible than spores in air. Spores trapped in solids have thermal resistance levels two or three times greater than those found for spores exposed to dry heat in air. Preliminary results on the combination of dry heat and ionizing radiation sterilization indicate no synergistic effects, i.e., the destruction obtained with each agent is additive. Another important variable that governs the interpretation of the effectiveness of dry heat sterilization tests is the recovery medium for heat damaged spores. The kinetics of dry heat sterilization cannot be fully interpreted from the available data. Death follows a logarithmic pattern thereby implying a monomolecular reaction. The mechanism of death is thought to be due to an oxidative process. PMID- 11883445 TI - Exposure of microorganisms to simulated extraterrestrial space ecology. AB - Spores of five test organisms, Bacillus subtilis var. niger, Bacillus megaterium, Bacillus stearothermophilus, Clostridium sporogenes and Aspergillus niger and soils were exposed, while under ultra high vacuum to temperatures of from -190 degrees C to +170 degrees C for 45 days. Up to a temperature of 25 degrees C no loss in viability of the test spores were noted when compared to original populations maintained at room temperature at atmospheric pressure over a desiccant. At elevated temperatures differences in resistivity occurred so that at 88 degrees C only B. subtilis var. niger and A. niger survived in appreciable numbers. At 107 degrees C only A. niger spores survived, but none were recoverable after exposure to 120 degrees C. In comparison B. subtilis var. niger survived at atmospheric pressure and 90 degrees C for 5 days while none of the other spores were viable after 48 hours. Four groups of organisms in soil, mesophilic aerobes and anaerobes, molds and actinomycetes were similarly tested in the vacuum chamber. From one soil sample only actinomycetes survived 120 degrees C, whereas one other soil sample yielded viable bacteria after exposure to 170 degrees C. The resistance of the isolates to high temperatures in the absence of soil is being evaluated. Several organisms have been found to resists 120 degrees C in ultra-high vacuum for 4-5 days, and experiments at higher temperatures are in progress. When irradiated with gamma rays from a cobalt-60 source, there was a difference between vacuum dried test spores irradiated while still under vacuum to those exposed to air immediately before irradiation. A reduction of from one-third to one-ninth of the viability of spores irradiated in vacuum occurred with vacuum-treated spores irradiated in air. PMID- 11883446 TI - A general review of chemical sterilization in space research. AB - The unsolved problems in the biology and the chemistry of chemical sterilization are legion. This discussion of them emphasizes some interesting areas for useful research. Studies in molecular biology and cancer chemotherapy have produced information which has important implications in chemical sterilization, particularly with the alkylating and the oxidizing agents. Many of these problems must be solved soon if chemical sterilization is to play the role it should in space research by providing means for effectively controlling the rate and extent of mingling of the forms of terrestrial life with those of extraterrestrial life. In space research in the United States, recent work on chemical sterilization has laid heavy emphasis on the identification and exploration of principles of engineering sciences which control sterilization effectiveness. These principles are reviewed and their significance in chemical sterilization is discussed in terms of the engineering factors involved. Among these factors are: consequence of sterilant polymerization or reaction with the object to be sterilized; criteria for chemical sterilization process effectiveness; depletion of the concentration of the sterilant by evaporation or chemical reaction, including polymerization; obstacles to the diffusion or convection of the sterilizing chemical to the immediate vicinity of the microorganisms to be destroyed; protection of the microorganisms from direct contact with the sterilant; phase behavior of the sterilant in the environment of the sterilization process; presence of water or water vapor; the nature, sources and numbers of microorganisms which survive standard chemical sterilization processes; and the length of exposure required to sterilize at each temperature. The significance of chemical sterilization to space science has been well reviewed at the previous COSPAR symposia. Because several thorough reviews of the empirical knowledge concerning the effects of chemical agents on microorganisms have been published recently, this information is not discussed again for chemical sterilants other than the alkylating agents and several of the oxidizing agents. This lack of emphasis indicates only that these other chemicals have not been applied to a large extent in space research to date and that discussions of their sterilizing properties are readily available in other recent reviews. In the future, interest in many of these other chemical sterilants will increase as the variety of devices to be sterilized and the variety of functions which sterilized devices are expected to perform increase. PMID- 11883447 TI - Problems in sterilization of unmanned space vehicles. AB - The probability of achieving and maintaining sterility of an unmanned spacecraft with varied suggested procedures is examined in detail, as are alternative techniques for avoiding biological contamination of the planets. The required degree of assurance against contamination of Mars, Venus and the Moon with Earth organisms is also considered. For Mars landers and orbiters, sterilization of the spacecraft or capsule by dry heat, with no subsequent access, is found to be desirable. For the present, sterilization of Venus landers still seems desirable. For Mars flybys, and for Venus orbiters and flybys, control of the trajectory to minimize the chance of unintentionally entering the planetary atmosphere appears the method of choice. For the Moon, sterilization seems unnecessary, but microbial counts should be kept low. Sterilization lowers spacecraft and system reliability. It reduces the chance of launching within periods fixed by astronomical constraints and increases costs. The gain which should be achieved through spacecraft sterilization, in return of significant biological information about the planets, must be balanced against these losses. In particular, one should keep in mind the loss in return of biological data occasioned by failure of a spacecraft to fulfill its mission; the probability of such failure is increased by sterilization. PMID- 11883448 TI - Gnotobiotic techniques and their application to spacecraft fabrication. AB - Several colonies of rats and mice have been reared totally free of viable microorganisms, some for over 10 years. A variety of other species including monkeys, dogs, sheep and swine have been maintained under the same conditions for shorter periods. This operation requires more precise control of microbic contamination than can be achieved by conventional techniques. The necessary control was realized by the use of apparatus termed "isolators". Obtaining sterility is a biological problem but its maintenance is an engineering problem. The gnotobiotic animal has been shown to be more sensitive to some microbic contamination than in vitro culture media. Apparently complex spacecraft cannot be sterilized effectively by postassembly treatment alone. Therefore, sterile subassemblies must be made as required with final assembly in a sterile environment to avoid trapping viable organisms. This can be accomplished within isolators since there appears to be no size limitations to the apparatus developed for biological and medical applications. Repairs and testing procedures under sterile conditions are possible with the isolator system. Fabrication of spacecraft within closed sterile isolators should present no major engineering problems provided all of the components are sterilizable. PMID- 11883449 TI - Fathers' grief when a disabled child dies. AB - This qualitative study examines the subjective experience of fathers' grief responses to the death of a child with a disability. Eight fathers were interviewed and completed the Grief Experience Inventory (GEI). GEI results indicated that fathers did not differ significantly from parents who lose a child in other ways. However, subjectively, fathers consistently reported that their bereavement was marked by a "double loss": disability and then death. Consistent with the literature on gender differences in bereavement, fathers reported greater emotional stoicism and used activity, rather than talk or social support, as a primary coping strategy. Clinical implications for professionals working with grieving men or with couples are discussed. PMID- 11883450 TI - Personal meanings of death in older adults and young adults in relation to their fears of death. AB - Age and gender differences in personal meanings of death have been noted from late childhood to middle adulthood but have been little studied in older adults, for whom death is less remote. Also, such meanings have not been related to their fears of death. Groups of 78 young adults (aged 19-29) and 68 older adults (aged 70-97) were compared on the Personal Meanings of Death (Extinction, After-life, Motivator, Legacy) and on 4 Multidimensional Fear of Death (MFODS) subscales. Analysis of variance indicated that the young scored higher (p <.05) than older adults on death as Motivator and on 3 MFODS subscales. Women scored higher than men on Motivator and fear of the dying process, but men had greater fear of the unknown. Death meanings After-life and Extinction were most strongly correlated with fear of death for both young and old. Further analysis revealed age differences within the older adult group. PMID- 11883451 TI - Roles for the church in improving end-of-life care: perceptions of Christian clergy and laity. AB - Although faith communities may seem to be logical places to discuss death and dying, few churches are engaged in extensive efforts to improve end-of-life care. To explore the meaning of a good death and potential roles for faith communities in this effort, the authors held focus groups involving 121 clergy and congregants affiliated with Christian churches in Honolulu. Participants' definitions of a good death were similar to those articulated in other studies, for example, pain is managed, inappropriate prolongation of dying is avoided, the family is present and supportive, conflicts are resolved, and spiritual/existential issues are addressed. Participants identified a number of roles for the church: (a) to help congregants prepare for death, both spiritually and practically; (b) to facilitate resolution of conflict and forgiveness; (c) to clarify if or how church theology should guide attitudes and practices related to death and dying; (d) to administer the appropriate rituals; and (e) to provide outreach to sick, dying, and bereaved members. A number of participants noted that attention to spiritual issues at life's end would be important to all people, and churches wanting to increase membership should expand offerings in this area. These findings suggest that faith communities can have a major impact on improving end-of-life care and that pastoral education include attention to these issues. PMID- 11883452 TI - Regional studies of homicide: a meta-analysis. AB - A meta-analysis of regional correlates of homicide rates in 8 nations revealed consistent associations of homicide rates with divorce, suicide and unemployment rates, per capita income, and population size. PMID- 11883453 TI - Data from human research projects will now be available to public. PMID- 11883454 TI - IRBs and their own conflicts of interest. PMID- 11883455 TI - Accreditation of entire human research protection programs. PMID- 11883456 TI - Institution must admit to parents that children should not have been subjects. PMID- 11883457 TI - Financial help for research subjects enrolled in "approved clinical studies". PMID- 11883458 TI - Accreditation and certification for human research protection programs. PMID- 11883459 TI - IRBs and financial conflict of interest. PMID- 11883463 TI - Intersexuality and the categories of sex. AB - Operations on intersexuals indicate that the sex of a person is based on more than biology. Expectations about proper gender activities furnish the frameworks through which certain features and combinations of features are understood to be fundamental to bodies and to comprise their sex. Yet, we can ask whether this interpretation is either coherent or consistent with our fuller conceptions of ourselves. Is there a point to interpreting a person as a sex? PMID- 11883465 TI - Coercion in the recruitment and retention of human research subjects, pharmaceutical industry payments to physician-investigators, and the moral courage of the IRB. PMID- 11883466 TI - Ethical implications of pediatric drug research policy initiatives. PMID- 11883467 TI - Resistant bacteria in retail meats and antimicrobial use in animals. PMID- 11883468 TI - Resistant bacteria in retail meats and antimicrobial use in animals. PMID- 11883469 TI - Transfusion in elderly patients with myocardial infarction. PMID- 11883470 TI - Transfusion in elderly patients with myocardial infarction. PMID- 11883471 TI - Infective endocarditis. PMID- 11883472 TI - Infective endocarditis. PMID- 11883473 TI - Unilateral osteoarthritis--"the working hand". PMID- 11883474 TI - An evaluation of human subjects protection at CDC/ATSDR. PMID- 11883475 TI - Selecting a surrogate to consent to medical research. PMID- 11883476 TI - Children as research subjects: new guidelines for Canadian IRBs. PMID- 11883477 TI - Declaration of Helsinki revised. PMID- 11883478 TI - Moral problems in assessing research risk. PMID- 11883479 TI - Moral solutions in assessing research risk. PMID- 11883480 TI - Prolonged weightlessness and calcium loss in man. AB - Data have been accumulated from a series of studies in which men have been subjected to weightlessness in orbital space flight for periods of up to 12 weeks. These data are used to predict the long term consequences of weightlessness upon the skeletal system. Space flight induced a loss of calcium which accelerated exponentially from about 50 mg/d at the end of 1 week to approx. 300 mg/d at the end of 12 weeks. The hypercalciuria reached a constant level within 4 weeks while fecal calcium losses continued to increase throughout the period of exposure. This apparent diminution of gastrointestinal absorptive efficiency was accompanied by a slight decline in the plasma level of parathyroid hormone and a slight elevation in the plasma level of calcium and phosphorus. Although losses in mineral from the calcaneus were closely correlated with the calcium imbalance, no changes were detected in the mineral mass of the ulna and radius. From the data presented it is concluded that the process of demineralization observed in space flight is more severe than would be predicted on the basis of observations in immobilized, bed rested, or paralyzed subjects. It is, moreover, suggested that the process may not be totally reversible. PMID- 11883481 TI - A review of the consequences of fluid and electrolyte shifts in weightlessness. AB - This review describes the renal-endocrine mechanisms related to the early losses of fluid-electrolytes from the body during weightlessness as well as their contribution to longer term adaptation of fluid-electrolyte balance. The hypotheses presented were generated by a systematic analysis of body fluid and renal dynamics observed under conditions of actual and simulated spaceflight. These have increased our understanding of the effects of acute headward fluid shifts on renal excretion, the factors promoting excess sodium excretion and the regulation of extracellular fluid composition. PMID- 11883482 TI - Monitoring the state of the human airways by analysis of respiratory sound. AB - A mechanism whereby sound is generated by the motion of vortices in the human lung is described. This mechanism is believed to be responsible for most of the sound which is generated both on inspiration and expiration in normal lungs. Mathematical expressions for the frequencies of sound generated, which depend only upon the axial flow velocity and diameters of the bronchi, are derived. This theory allows the location within the bronchial tree from which particular sounds emanate to be determined. Redistribution of pulmonary blood volume following transition from Earth gravity to the weightless state probably alters the caliber of certain airways and doubtless alters sound transmission properties of the lung. We believe that these changes can be monitored effectively and non invasively by spectral analysis of pulmonary sound. PMID- 11883483 TI - Demystifying central review boards: current options and future directions. PMID- 11883484 TI - Research ethics and the "fieldwork monitoring committee". PMID- 11883485 TI - Food research and the IRBs. PMID- 11883486 TI - The IRB is not a data and safety monitoring board. PMID- 11883487 TI - Preventive medicine. PMID- 11883488 TI - Expressions of the spirit in Catholic medical practice. PMID- 11883489 TI - The utility of embryonic stem cells. PMID- 11883490 TI - Human gene transfer: some theological contributions to the ethical debate. PMID- 11883491 TI - The stem cell dilemma--an overview. PMID- 11883492 TI - On synthetic life. PMID- 11883493 TI - Retraction. PMID- 11883494 TI - The influenza vaccine. PMID- 11883495 TI - Assessing the educational needs and concerns of nursing home staff regarding end of-life care. AB - The purpose of this study was to explore and describe the educational needs and concerns of licensed nursing staff and certified nursing assistants (CNAs) regarding end-of-life (EOL) care. Focus group interviews were conducted at two nursing homes in the Pacific Northwest. Separate interviews were conducted for licensed staff (RNs and LPN/LVNs) and CNAs. A total of 15 licensed staff and 39 CNAs participated in the study. Interviews were transcribed and themes were extracted through consensus reached by three investigators. The major concerns of these nursing home staff focused on symptom management, communication and interactions, goals of care, role delineation, time constraints, self-care needs, and emotional attachment to residents. Although both groups described similar themes, specific issues within each topic often were different for licensed staff and CNAs. These findings can be used to guide the design of educational programs aimed at assisting nursing home staff in providing high level end-of-life care. PMID- 11883496 TI - Advance directives and dementia. AB - Since the 1990 Patient Self-Determination Act, increasing numbers of adults are completing advance directives (ADs), but unfortunately many adults seen in a dementia evaluation program have not completed an AD. This article discusses the issue of individuals with dementia completing ADs. Situational factors that frame this issue are the stage of dementia, degree of certainty of an individual's wishes for end-of-life care, the decision-making act required by care providers, and the degree of contentment or distress experienced by an individual with dementia. Several investigators have demonstrated successful completion of ADs by individuals with mild and moderate dementia. A nurse's knowledge about the stages of dementia is essential to helping an individual through the AD decision-making process. Nurses caring for individuals with dementia should assess decision making context; recognize the emotions of family, friends, and staff; understand the substance and logic of AD requests; and support individuals and their decisions. PMID- 11883497 TI - The effects of bathing and skin care practices on skin quality and satisfaction with an innovative product. PMID- 11883498 TI - Community needs assessment for nursing homes: a Taiwan study. AB - The assessment of nursing home needs is becoming an important research endeavor because of the rapid social changes in the past 20 to 30 years in Taiwan. The purpose of this article is to present the methodology, as well as the results of a nursing home community needs assessment. A cross-sectional descriptive study using a mailed questionnaire was conducted. The results indicated low awareness level, low personal experience, high service needs, and high potential for use of nursing home services among the individuals living in 10 surveyed towns (N = 319). PMID- 11883499 TI - Rethinking advanced directives. PMID- 11883500 TI - How do you structure the day for individuals with dementia? PMID- 11883501 TI - Hydration management protocol. PMID- 11883502 TI - Phage display as a tool for protease ligand discovery. AB - Proteolytic enzymes have been implicated as the pathological agent in a number of disease states. For this reason proteases are attractive therapeutic targets. Phage display of peptide libraries can be used to identify peptides that may be used either directly as inhibitors or serve as leads in the generation of prodrugs and peptidomimetics. PMID- 11883503 TI - Direct selection of cDNAs from filamentous phage surface display libraries: potential and limitations. AB - Over the past decade, powerful technologies devoted to survey large molecular libraries for the presence of specific clones using the discriminative power of affinity selection have been developed. Phage surface display technology is the most established of these methods and has revolutionised our ability to select agonistic and antagonistic peptides, antibodies with desired specificity and other drug targets. Thereby ligands are expressed as fusions to phage coat proteins and their respective genes are packaged within the phage. The basic concept of linking the phenotype, expressed as gene product displayed on the phage coat, to its genetic information integrated into the phage genome, creates fusion proteins covalently associated with the infectious particle itself. Binding of the phage to the target molecule offers a selective system by which rare phage carrying the desired gene product can be selected from large phage populations carrying inappropriate sequences. Phage selected in this fashion can be used for subsequent rounds of selection because they are able to self replicate in suitable host cells, yielding target-specific phage populations after several consecutive rounds of affinity selection. Over 1500 publications describe the use of phage display technology highlighting its performance. Phage display possesses certain limitations, including its use for selection of genes from cDNA libraries that has lagged behind, despite the many accomplishments of this technology. Here we discuss recent progress in construction and screening of cDNA libraries displayed on phage surface and emerging concepts allowing fast identification of virtually all different clones present in enriched libraries. PMID- 11883504 TI - Bacteriophage lambda as a vehicle for peptide and protein display. AB - Bacteriophage lambda has emerged as an alternative vehicle for the surface display of peptides and proteins to the commonly used filamentous phage. There are a number of unique features that make lambda an attractive display vehicle including the ability to display multimeric proteins, no requirement for secretion of the displayed fusion protein and the means to vary the valency of the displayed fusion protein. With its increasing use for cDNA encoded display, the lambda display systems will be an important tool for functional genomics. PMID- 11883505 TI - Application of phage display technology to cancer research. AB - Despite years of international effort, cancer remains a major cause of death in developed countries, claiming more than 500000 lives per year in the United States alone. Recombinant DNA technology and high throughput screening methods have recently increased the pace of cancer research. In this review, we will examine the impact and contribution of phage display technology to this area of research. As a biological combinatorial system, the strength of phage display lies in its flexibility and its ability to efficiently study protein-protein interactions. The technology has also facilitated the discovery of molecules that have potential roles in the diagnosis and treatment of cancer. PMID- 11883506 TI - Evolving phage vectors for cell targeted gene delivery. AB - We adapted filamentous phage vectors for targeted gene delivery to mammalian cells by inserting a mammalian reporter gene expression cassette (GFP) into the vector backbone and fusing the pIII coat protein to a cell targeting ligand (i.e. FGF2, EGF). Like transfection with animal viral vectors, targeted phage gene delivery is concentration, time, and ligand dependent. Importantly, targeted phage particles are specific for the appropriate target cell surface receptor. Phage have distinct advantages over existing gene therapy vectors because they are simple, economical to produce at high titer, have no intrinsic tropism for mammalian cells, and are relatively simple to genetically modify and evolve. Initially transduction by targeted phage particles was low resulting in foreign gene expression in 1-2% of transfected cells. We increased transduction efficiency by modifying both the transfection protocol and vector design. For example, we stabilized the display of the targeting ligand to create multivalent phagemid-based vectors with transduction efficiencies of up to 45% in certain cell lines when combined with genotoxic treatment. Taken together, these studies establish that the efficiency of phage-mediated gene transfer can be significantly improved through genetic modification. We are currently evolving phage vectors with enhanced cell targeting, increased stability, reduced immunogenicity and other properties suitable for gene therapy. PMID- 11883507 TI - Possible mechanism of anticonvulsant effect of ketamine in mice. AB - The study was designed to investigate the effect of ketamine on convulsive behaviour using maximal electroshock (MES) test. An attempt was also made to study the possible receptor mechanisms involved. MES seizures were induced in mice via transauricular electrodes (60 mA, 0.2sec). Seizure severity was assessed by the duration of tonic hindlimb extensor phase and mortality due to convulsions. Intraperitoneal administration of ketamine produced a dose-dependent (5-50 mg/kg) protection against hindlimb extensor phase. The anticonvulsant effect of ketamine was antagonized neither by naloxone (low as well as high doses) nor sulpiride, but was attenuated by haloperidol, a dopamine (D2)/sigma receptor antagonist. Co-administration of gamma-aminobutyric acid (GABA)-ergic drugs (GABA, muscimol, diazepam and baclofen) and N-methyl-D-aspartate (NMDA) receptor antagonist, dizocilpine (MK801) with ketamine facilitated the anticonvulsant action of the latter drug. In contrast, flumazenil, a benzodiazepine (BZD)-GABAA receptor antagonist, reversed the facilitatory effect of diazepam on the anti-MES effect of ketamine. Similarly, delta-aminovaleric acid (DAVA), antagonized the facilitatory effect of baclofen on anti-MES action of ketamine. These BZD-GABAergic antagonists, flumazenil or DAVA per se also attenuated the anti-MES effect of ketamine given alone. The results suggest that besides its known antagonistic effect on NMDA channel, other neurotransmitter systems i.e. sigma, GABAA-BZD-chloride channel complex and GABAB receptors may also be involved in the anti-MES action of ketamine. PMID- 11883508 TI - Modulation by insulin rather than blood glucose of the pain threshold in acute physiological and flavone induced antinociception in mice. AB - The present study investigated the cause effect relationship between glycemic and algesic states. The hypo- and hyperglycemic conditions were induced physiologically through exercise (3 min swim at room temperature 28 degrees - 30 degrees C) and external dextrose (2 g/kg, ip) administration respectively in mice. Besides, flavone (50 mg/kg, sc) a known antinociceptive drug was chosen to study such a cause effect relationship. The anti-nociception was assessed by acetic acid assay, blood glucose measured using glucometer (Ames) and serum insulin by radioimmunoassay. The findings revealed that irrespective of the glycemic state whether hypo-, hyper, or euglycemic induced by swim stress, dextrose or flavone per se respectively, significant antinociceptive response was recorded. Pretreatment with flavone (50 mg/kg, sc) always exhibited a tendency to reverse the hyperglycemia, if any, but enhanced the antinociceptive response either after swim stress or after dextrose. These data support the contention that changes in the glycemic state in acute condition is not responsible for antinociceptive response and thereby suggesting dissociation between these two parameters. Extended studies estimating serum insulin level after the above mentioned maneuvers showed a significant rise whenever antinociceptive response was recorded irrespective of the glycemic state. It is suggested that serum insulin level, a hormonal parameter rather than the blood glucose level, which is a metabolic parameter, appears more reliable. It appears that the changes in serum insulin level produced by various treatments may have a relationship with the antinociceptive response. However, this study has the limitation that the results can apply only for acute conditions and extrapolation to clinical conditions is debatable. PMID- 11883509 TI - Effects of endosulfan on intestinal functions in protein-malnourished rats. AB - In rats fed 18% protein diet, administration of endosulfan (2mg/kg body weight daily for 7 days) significantly decreased the brush border sialic acid and increased the hexoses contents. The intestinal uptake of glucose was increased while that of glycine and calcium was reduced. Brush border enzymes and lipids were not affected. However, in protein malnourished rats (fed 8% protein) exposed to endosulfan, brush border sucrase and peptidase activities were enhanced, while alkaline phosphatase activity was decreased compared to untreated malnourished animals. Membrane sialic acid content was low while fucose and cholesterol levels were augmented in endosulfan fed malnourished animals. The uptake of glucose and glycine was elevated under these conditions. These results Suggest that the nutritional status of the animals has an important bearing on thc susceptibility of intestinal tissue to endosulfan toxicity in rats. PMID- 11883510 TI - Hepatoprotective action of abhrak bhasma, an ayurvedic drug in albino rats against hepatitis induced by CCl4. AB - Abhrak bhasma is a commonly used ayurvedic drug against many diseases including hepatitis. It is tested in albino rats using a model of hepatitis induced by a single dose of CCl4 (3 ml/kg body wt). Different doses of abhrak bhasma (10, 20, 30 and 40 mg/kg body wt) were tested to decide the dose related hepatoprotective efficacy. The centrolobular necrosis induced by single dose of CCl4 was reduced significantly by abhrak bhasma (10 mg) and liver histology was also protected by 20 mg dose. Liver acid lipase activity was lowered, while alkaline and lipoprotein lipase activities were elevated due to treatment of single dose of CCl4. Abhrak bhasma counteracted the action of CCl4 on liver lipolytic enzymes. CCl4 did not alter the kidney histologically. Activities of three lipases of rat kidney (acid, alkaline and lipoprotein lipases) were reduced by CCl4 treatment and were reversed by administration of abhrak bhasma. Acid lipase activity of rat adipose tissue was reduced by CCl4 treatment. On the contrary alkaline, lipoprotein and hormone sensitive lipases were enhanced after 24 hr of administration of CCl4. Acid lipase activity was raised by administration of different doses of abhrak bhasma concurrent with CCl4. Abhrak bhasma treatment along with CCl4 enhanced alkaline lipase activity at 10 and 20 mg dose and later it was reduced at 30 and 40 mg doses and came to normal levels. Lipoprotein and hormone sensitive lipases were reduced by the counteraction of increasing doses of abhrak bhasma. PMID- 11883511 TI - Hypolipidemic and antiperoxidative effect of coconut protein in hypercholesterolemic rats. AB - Effect of coconut protein in rats fed high fat cholesterol containing diet on the metabolism of lipids and lipid peroxides was studied. In addition, effect of coconut protein were compared with rats fed L-arginine. The results indicate that those fed coconut protein and those fed L-arginine showed significantly lower levels of total cholesterol, LDL+ VLDL cholesterol, Triglycerides and Phospholipids in the serum and higher levels of serum HDL cholesterol. The concentration of total cholesterol, triglycerides and phospholipids in the tissues were lower in these groups. There was increased hepatic cholesterogenesis which is evident from the higher rate of incorporation of labeled acetate into free cholesterol. Increased conversion of cholesterol to bile acids and increased fecal excretion of bile acids were observed. Feeding coconut protein results in decreased levels of Malondialdehyde in the heart and increased activity of Superoxide dismutase and Catalase. Supplementation of coconut protein causes increased excretion of urinary nitrate which implies higher rate of conversion of arginine into nitric oxide. In the present study, the arginine supplemented group and the coconut protein fed group produced similar effects. These studies clearly demonstrate that coconut protein is able to reduce hyperlipidemia and peroxidative effect induced by high fat cholesterol containing diet and these effects are mainly mediated by the L-arginine present in it. PMID- 11883512 TI - Interconversion of free sugars in relation to activities of enzymes catalyzing synthesis and cleavage of sucrose in growing stem tissues of sorghum. AB - Free sugar interconversion and activities of soluble acidic (pH 4.8) and neutral (pH 7.5) invertases, sucrose synthase (synthesis) and sucrose phosphate synthase were investigated in the growing nodes and internodes of sorghum (Sorghum vulgare). The results were substantiated with incorporation of 14C from supplied sucrose and hexoses into endogenous sugars of these stem tissues. With the advancement in plant growth, the content of total free sugars in apical nodes and internodes increased till 70 DAS (flowering stage) followed by a decline. In the corresponding basal tissues, the sugar build-up continued even beyond this stage of plant growth. Compared with basal stem tissues, the apical ones contained high activities of soluble invertases and a low proportion amongst free sugars of sucrose. The activities of sucrose-hydrolyzing enzymes were higher as compared with those of sucrose-synthesizing ones in both nodes and internodes and with the growth of plant, the activity of neutral invertase increased in these tissues. More 14C from supplied sucrose and hexoses appeared in extracted sugars from cut discs of apical nodes and internodes in comparison with their basal counterparts. 14C from supplied sucrose appeared in glucose, fructose and from supplied hexoses appeared in sucrose. The results suggest that in apical nodes and internodes, where a rapid cell division and cell expansion occur, sucrose is obligatorily inverted to meet the increased requirement of hexoses and there is a compartmentalized synthesis and cleavage of sucrose in the nodes and internodes of growing sorghum plant. PMID- 11883513 TI - Cultural requirements for in vitro seed germination, protocorm growth and seedling development of Geodorum densiflorum (Lam.) Schltr. AB - Effects of different nutrient solutions, organic supplements and plant growth regulators on in vitro seed germination and protocorm development of Geodorum densiflorum (Lam.) Schltr. were studied. Seed germination was very high (up to 96%) in all the basal media, with Knudson's C and half-strength Murashige & Skoog being slightly more productive than Vacin & Went. Application of organic supplements and NAA had little effect on germination, but BAP proved inhibitory. After germination, protocorms exhibited a clear preference for peptone and NAA for much faster growth, while BAP resulted in stunted growth. Beside normal development, disorganisation of protocorms, followed by callusing occurred in presence of peptone and NAA. The calli were compact with limited growth and frequently regenerated protocorm like bodies. Development of seedlings was preceded by an intermediary rhizome phase. Growth of rhizomes was slow in the plant growth regulator free medium and about 15 months of culture was required for seedling formation. However, it was possible to hasten the process by 8-10 months with the employment of NAA, which also enhanced the number of seedlings per protocorm through axillary branching. Combined application of high BAP and low NAA was also useful for high rate of seedling formation. PMID- 11883514 TI - Surface characters and extracellular toxins involved in the pathogenesis of Aeromonas hydrophila. AB - A small number of serotypically distinct strains of A. hydrophila obtained from diseased freshwater fish were examined for their pathogenic properties comprising of cell surface characteristics and extracellular toxins. Test strains exhibited homogeneity in their cell surface characteristics despite being serologically heterogeneous. Studies on extracellular biological activities revealed qualitative and quantitative differences in production of toxins, probably explaining their antigenic diversity. Three distinct proteases, namely heat stable metallo protease, heat labile serine protease and heat labile metallo protease were identified from the strains. PMID- 11883515 TI - Effect of phenol on protein and amino acid content of Xanthomonas oryzae pv. oryzae. AB - Leaf blight disease of rice (Oryza sativa) is caused by the bacterium Xanthomonas oryzae pv. oryzae. Phenol (1 to 4 mM) induced changes in protein profiles of X. o. pv. oryzae and a stress protein with a molecular mass of 69,000 appeared. HPLC analysis indicated occurrence of amino acids such as asparagine, alanine, methionine and cystine in phenol treated cells. Proton NMR analysis also revealed variation on the presence of amino acids in the cells treated with phenol. PMID- 11883516 TI - Effect of phenol on lipid and fatty acid profile of Xanthomonas oryzae pv. oryzae. AB - Effect of phenol on total lipid and fatty acid composition of Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight of rice (Oryzae sativa) was studied. Lipid level was low in phenol treated cells. Number of fatty acids detected from phenol treated cells was more than those found in untreated cells as revealed by Gas chromatography. Pentadecanoic acid (C15:0), linolenic acid (C18:3) and behenic acid (C22:0) were present only in the treated cells. Palmitic acid which is usually found in bacteria was not detected both in control and treated cells. PMID- 11883517 TI - Characterization of toxin from cheilanthes fern and its effect on lymphocyte proliferation and DNA fragmentation. AB - Thin layer chromatography of aqueous extract of whole Cheilanthesfarinosa fern indicated the presence of ptaquiloside or ptaquiloside like compound, coinciding Rf values with that of Pterosin B standard. HPLC analysis revealed the presence of 26.3 mg/kg ptaquiloside. In vitro studies of the aqueous extract on lymphocyte culture revealed a correlation between stimulative indices and concentration of aqueous extract. Stimulation in lymphocyte proliferation was in order of bracken > cheilanthes > ConA> ptaquiloside standard. On incubation of lymphocyte with aqueous extract of ferns, no DNA damage was observed in isolated DNA. PMID- 11883518 TI - Clastogenic effects of dietary supplement--Spirulina alga, and some medicinal plant products from Boswellia serrata, Withania somnifera on mice. AB - Pretreatment of aqueous extracts of Zyrulina (Spirulina), Aswagandha (Withania) and Nopane (Boswellia) on colchicine induced chromosome damage showed weakness of clastogenic activity in Swiss albino mice. None of the treatments increased significantly the number of chromosome aberrations. PMID- 11883519 TI - Human genome project: pharmacogenomics and drug development. AB - Now that all 30,000 or so genes that make up the human genome have been deciphered, pharmaceutical industries are emerging to capitalize the custom based drug treatment. Understanding human genetic variation promises to have a great impact on our ability to uncover the cause of individual variation in response to therapeutics. The study of association between genetics and drug response is called pharmacogenomics. The potential implication of genomics and pharmacogenomics in clinical research and clinical medicine is that disease could be treated according to the interindividual differences in drug disposition and effects, thereby enhancing the drug discovery and providing a stronger scientific basis of each patient's genetic constitution. Sequence information derived from the genomes of many individuals is leading to the rapid discovery of single nucleotide polymorphisms or SNPs. Detection of these human polymorphisms will fuel the discipline of pharmacogenomics by developing more personalized drug therapies. A greater understanding of the way in which individuals with a particular genotype respond to a drug allows manufacturers to identify population subgroups that will benefit most from a particular drug. The increasing emphasis on pharmacogenomics is likely to raise ethical and legal questions regarding, among other things, the design of research studies, the construction of clinical trials and the pricing of drugs. PMID- 11883520 TI - Leprosy bacillus--possibly the first chemoautotrophic human pathogen cultivated in vitro and characterised. AB - Leprosy bacillus (LB) and leprosy derived in vitro culture forms, the chemoautotrophic nocardioform (CAN) bacteria, showed an extremely close homology and identity with each other as regards a chemoautotrophic nutritional pattern, a nocardioform morphology, a weak acid-fastness coupled with Gram and Gomori's stain positivity, an exclusive mycolate and lipid profile, a phenolic glycolipid (PGL-I) and a highly sequestrated DNA characteristic, namely, a unique small size, a low G+C % mole, an exceptionally high gamma and UV radiation resistance, and a high thermal resistance. LB/CAN bacteria (CANb) gave positive signals for 36 kDa protein PCR, as well as, for 65 kDa epitope, and hybridisation with two or more probes and also by RFLP-analysis. Both LB/and CAN bacteria exhibited bacillary multiplication in the mouse footpads (MFP), nerve infiltration and evidences for local pathogenicity associated with pronounced systemic invasion. A highly reproducible mutilation model could be established which enabled a successful application of the postulates of Koch. The proof of their total identity was their anergic reactions in LL cases counterpoised against Mitsuda type strong nodular responses, mirroring the reactions of leprosy bacilli in TT cases, in accordance with the dictum of XIth International Leprosy Congress (1978). Thus, the chemoautotrophic nutritional requirements of LB, entirely unsuspected for a medically important pathogenic bacterium, having dimorphic (both bacillary and mycelial) characters with spores, mycelia and granules and unique pathogenicity of multilation manifested through the virulence factor, the enzyme collagenase, made LB or M leprae the highly enigmatic bacterium for so long. PMID- 11883521 TI - A dot-immunobinding assay (dot-Iba) for rapid diagnosis of pulmonary tuberculosis. AB - IgG antibody to Mycobacterium tuberculosis from the sera of patients with 'definite' pulmonary tuberculosis (PT) was isolated and coupled with Cyanogen bromide-Sepharose 4B. Using an immunoabsorbent affinity chromatography, 14 kDa antigen was recovered from the culture filtrates of M. tuberculosis. With this mycobacterial antigen, a dot immunobinding assay (Dot-Iba) was developed for the detection of specific antibody to M. tuberculosis in the sera of patients with PT and controls. The assay gave positive results in all the 12 sputum-smear positive [acid fast bacilli (AFB)] patients with PT and gave negative results in the 50 sera from control groups. The Dot-Iba as described in this study, is simple, rapid and specific for laboratory diagnosis of PT. PMID- 11883522 TI - Mycobacterium phlei as an oral immunomodulator with Newcastle disease vaccine. AB - Experiments were conducted in chickens to understand the effects of oral immunomodulation. Heat inactivated M phlei, a commensal Mycobacterium and a non specific immunomodulator, was administered orally prior to live Newcastle disease F (ND F) strain vaccination. In experimental birds it lead to an enhanced cell mediated Immune response (CMI) against the vaccine. There was a reduction in the Haemagglutination inhibiting (HI) antibodies. However, it did not affect the protection against a virulent challenge, as the protection percentage was more or less same in vaccinated birds irrespective of the M.phlei administration. M. phlei administration could not enhance the immune response to inactivated ND F vaccine administered orally. The results indicate that M. phlei favours a CMI response to orally administered live ND F vaccine. It may be of potential use in enhancing CMI against vaccines and a cheaper alternative to costlier recombinant cytokines. PMID- 11883523 TI - Targetted localisation and imaging of a murine lymphoma using 131I-labelled monoclonal antibody. AB - In vivo tumor targetting with radiolabelled monoclonal antibodies is a promising approach for the diagnosis and therapy of tumors. A specific monoclonal antibody (mAb), DLAB was generated to the Dalton's lymphoma associated antigen (DLAA) from Haemophilus paragallinarum-induced spontaneous fusion. In order to study the tumor localisation and biodistribution properties of the monoclonal antibody, scintigraphic studies were performed using the radiolabelled DLAB. 131-labelled DLAB was administered intravenously into Swiss mice bearing Dalton's lymphoma and external scintiscanning was performed at different time intervals. Clear tumor images were obtained which revealed selective and specific uptake of radiolabel and the results were compared with biodistribution data. The radioiodinated monoclonal antibody showed fast tumor uptake which increased significantly to 14.6% injected dose (ID)/g at 12 hr post-injection. Enhanced blood clearance of radioactivity resulted in higher tumor/blood ratio of 5.96 at 48 hr. 131I labelled DLAB resulted in selective and enhanced uptake of the radioactivity by the tumor compared to the non-specific antibody and the results suggest the potential use of spontaneous fusion for producing specific monoclonal antibodies for tumor detection and therapy. PMID- 11883524 TI - Interaction of flunarizine with sodium valproate or ethosuximide in gamahydroxybutyrate induced absence seizures in rats. AB - Sodium valproate(VPA), ethosuximide(ESM), 200 mg/kg ip and flunarizine (FLU) 5 or 10 mg/kg ip were first administered independently to rats in order to study their effects on behavioural and EEG aspects of spike and wave discharges (SWDs) induced by y- hydroxybutyrate (GHB,100 mg/kg ip). GHB treated rats show behavioural changes and concomitant repetitive EEG episodes of 7 to 9 Hz SWDs, mimicking human absence seizures (AS), and can be used as a pharmacological model. The number and duration of SWDs were calculated for 1 hr from the EEG and were parameters for drug evaluation. VPA and ESM at 200 mg/kg, significantly reduced SWD number and duration/hr, while FLU showed significant reduction only at 10 but not at 5 mg/kg. Combination of FLU, 10 mg/kg with either VPA or ESM showed significant reduction of SWD number and duration, suggesting an additive effect of the anti-absence agents with the calcium channel blocker, FLU, on experimental absence seizures in rats. PMID- 11883525 TI - Gene expression in inherited breast cancer. AB - Large proportions of hereditary breast cancers are due to mutations in the two breast cancer susceptibility genes BRCA1 and BRCA2. Considerable effort has gone into studying the function(s) of these tumor suppressor genes, both in attempts to better understand why individuals with these inherited mutations acquire breast (and ovarian) cancer and to potentially develop better treatment strategies. The advent of tools such as cDNA microarrays has enabled researchers to study global gene expression patterns in, for example, primary tumors, thus providing more comprehensive overviews of tumor development and progression. Our recent study (Hedenfalk et al., 2001) strongly supports the principle that genomic approaches to classification of hereditary breast cancers are possible, and that further studies will likely identify the most significant genes that discriminate between subgroups and may influence prognosis and treatment. A large number of hereditary breast cancer cases cannot be accounted for by mutations in these two genes and are believed to be due to as yet unidentified breast cancer predisposition genes (BRCAx). Subclassification of these non-BRCA1/2 breast cancers using cDNA microarray-based gene expression profiling, followed by linkage analysis and/or investigation of genomic alterations, may help in the recognition of novel breast cancer predisposition loci. To summarize, gene expression-based analysis of hereditary breast cancer can potentially be used for classification purposes, as well as to expand upon our knowledge of differences between different forms of hereditary breast cancer. Initial studies indicate that a patient's genotype does in fact leave an identifiable trace on her/his cancer's gene expression profile. PMID- 11883526 TI - Genetic requirements for the episomal maintenance of oncogenic herpesvirus genomes. AB - Herpesviruses are large double-stranded DNA viruses that are characterized by lifelong latency. Epstein-Barr virus (EBV), the recently discovered Kaposi's sarcoma associated herpesvirus (KSHV), also referred to as human herpesvirus-8 (HHV-8), and the simian Herpesvirus saimiri (HVS) are associated with malignant lymphoproliferative diseases. These viruses establish latent infection in lymphoid cells. During latency only a few viral genes are expressed and the viral genome persists as a multicopy circular episome. The episome contains repetitive sequences that serve as multiple cooperative binding sites for the viral DNA binding proteins Epstein-Barr virus nuclear antigen 1 (EBNA-1) of EBV and latency associated nuclear antigen (LANA1) of KSHV and HVS, which are expressed during latency. The oligomerized proteins associate with the viral genome and tether it to host chromosomes, assuring continual lifelong persistence of the virus. PMID- 11883527 TI - Treatment of Epstein-Barr virus-associated malignancies with specific T cells. AB - Latent Epstein-Barr virus (EBV) infection is associated with a heterogeneous group of malignancies, including Burkitt's lymphoma, Hodgkin's disease, nasopharyngeal carcinoma, and lymphoproliferative disease (LPD). The development of adoptive immunotherapies for these malignancies is being fueled by the successful generation of allogeneic donor derived EBV-specific cytotoxic T cells (CTL) for the prevention and treatment of EBV-LPD after hematopoietic stem cell transplantation. This approach is being extended to EBV-LPD after solid organ transplantation by use of autologous and haploidentical EBV-specific CTL. For other EBV-associated malignancies, there is only limited clinical experience with EBV-specific CTL. With few exceptions, only patients with recurrent Hodgkin's disease have been treated with autologous EBV-specific CTL, and although the results have been promising, they do not include cures. Lack of CTL efficacy may reflect either down-regulation of immunodominant EBV proteins, which are major CTL targets, or the presence of inhibitory cytokines. Further improvement of EBV specific CTL therapy for Hodgkin's disease will require improved methods to activate and expand CTL specific for the latent EBV genes expressed in Hodgkin's disease and to genetically modify the expanded CTL to render them resistant to inhibitory cytokines. If effective, such strategies could be applied not only to other EBV-associated malignancies, but also to a broad range of human tumors with defined tumor antigens and similar immune evasion strategies. PMID- 11883528 TI - Role of glycogen synthase kinase-3 in cancer: regulation by Wnts and other signaling pathways. AB - Although glycogen synthase kinase-3 (GSK-3) is but one of more than a thousand distinct serine/threonine kinases present in the mammalian genome, this enzyme has attracted attention for its role in a diverse range of cellular processes and its positioning at a nexus of several signaling pathways that are important in cancer and other human diseases. The association of GSK-3 with widely different functions, from glycogen metabolism to fruit fly segmentation and slime mold differentiation, was initially perplexing. However, as the context of the biological processes involving this enzyme has been clarified, unifying themes have emerged that begin to explain its pleiotropic nature. Unlike most protein kinases involved in signaling, GSK-3 is active in unstimulated, resting cells. Its activity is inactivated during cellular responses and its substrates therefore tend to be dephosphorylated. As more of these targets have been identified and the effects of their modification by GSK-3 determined, most have been found to be functionally inhibited by GSK-3. Hence, this kinase appears to act as a general repressor, keeping its targets switched off or inaccessible under resting conditions. The rarity of this form of regulation is perhaps related to the diversity of its targets. Over the past decade, the importance of GSK-3 has been established by three significant properties: its remarkable evolutionary conservation, allowing analysis in genetically tractable organisms; its involvement in the Wnt/wingless signaling pathway; and its inhibition by agonists of the prosurvival phosphatidylinositol 3' kinase (P13'K) pathway. This review covers recent advances in understanding the physiological roles of this enzyme, particularly in the context of cancer. PMID- 11883529 TI - Chronic immune activation and inflammation in the pathogenesis of AIDS and cancer. AB - Infection with the human immunodeficiency virus (HIV) invariably leads to the development of acquired immunodeficiency syndrome (AIDS) in most infected humans, yet does so rarely, if at all, in HIV-infected chimpanzees. The differences between the two species are not due to differences in cellular receptors or an inability of the chimpanzee to be infected, but rather to the lack of pan-immune activation in the infected primate. This results in reduced apoptotic death in CD4+ T-helper lymphocytes and a lower viral load. In humans the degree of chronic immune activation correlates with virus load and clinical outcome with high immune activation leading to high viral loads and the more rapid progression to AIDS and death. The type of immune perturbation seen in HIV-associated AIDS is similar to that of chronic graft-versus-host disease (GVHD) where reduced cell mediated immune (CMI) responses occur early in the course of the disease and where humoral responses (HI) predominate. A reduced CMI response occurs in a number of chronic infectious diseases, including tuberculosis and leishmaniasis. More recently, it has become increasingly apparent that the CMI response is suppressed in virtually all malignant diseases, including melanoma and colorectal and prostate cancer. This raises the possibility that, as the malignant process develops, the cancer cells evolve to subvert the CMI response. Moreover, the reduced CMI response seen in colorectal cancer (CRC) patients is completely reversed following curative surgery strongly supporting the hypothesis that CRC can suppress the systemic immune response. Wound healing, ovulation, embryo implantation, and fetal growth are all associated with suppressed CMI and neovascularization (the formation of new blood vessels) or angiogenesis (the formation of new blood vessels from an existing vasculature). If unresolved, wound healing results in chronic inflammation, which can give rise to the phenomenon of "scar cancers." Indeed all the chronic inflammatory conditions known to be associated with the subsequent development of malignant disease, including chronic obstructive airway disease (COPD), ulcerative colitis (UC), and asbestosis, give rise to similar proangiogenic, suppressed CMI, and HI predominant environments. In keeping with this CMI-associated cytokines such as interleukin (IL)-2 and interferon (IFN)-gamma tend to be antiangiogenic, whereas HI cytokines such as IL-6 tend to be proangiogenic. Furthermore, chronic immune activation leads to the synthesis and release of factors such as macrophage inflammatory protein (MIP)-1 that inhibit apoptosis through suppression of p53 activity. The "Golden Triangle" of suppressed CMI, angiogenesis, and reduced apoptosis would provide the ideal environment for the serial mutations to occur that are required for the development of malignant disease. If the observed association is relevant to carcinogenesis, then treatments aimed at reducing the components of these inflammatory conditions may be useful both in the setting of chemoprevention and the therapeutic management of established disease. PMID- 11883530 TI - Molecular biology of Hodgkin's lymphoma. AB - Hodgkin's lymphoma (HL) is characterized by typical mononucleated Hodgkin and multinucleated Reed-Sternberg cells, which occur at low frequency in a mixed cellular infiltrate in the tumor tissue. Because of the rarity of these cells and their unusual immunophenotype, which is strikingly different from those of all normal hematopoietic cell types, the origin of these cells and their clonality have long been unclear. Single-cell studies of rearranged immunoglobulin genes showed that Hodgkin and Reed-Sternberg (HRS) cells represent clonal tumor-cell populations derived from germinal center B cells. In classical HL, the detection of obviously crippling immunoglobulin gene mutations in a fraction of the cases suggests that HRS cells may derive from germinal center B cells that have lost the capacity to be positively selected by antigen and that normally would have undergone apoptosis. In rare cases, HRS cells represent transformed T lymphocytes. The transforming events involved in malignant transformation of HRS cells are still largely unknown. Constitutive activation of the transcription factor NFkappaB, which can, for example, be induced through Epstein-Barr virus transformation of HRS cells or destructive somatic mutations of the inhibitor of NFkappaB, is likely to be a key event in HL pathogenesis. Significant progress has been made in our understanding of the cellular interactions in HL tissues, which are mainly mediated by a large variety of cytokines and chemokines. PMID- 11883531 TI - Multiparameter analyses of cell cycle regulatory proteins in human breast cancer: a key to definition of separate pathways in tumorigenesis. AB - Breast cancer is one of the most common cancer forms affecting many women. The disease nevertheless has widely varying behavior and therefore patient outcome, and an important undertaking is to define and understand the molecular mechanisms behind these actions. Defects in the G1/S transition in the cell cycle affect both tumor proliferation and the fidelity of check points responsible for chromosomal integrity and DNA damage response and has lately been shown to represent one of a rather limited set of key aberrations in the transformation process. Many cell cycle regulatory proteins are either oncogenes or suppressor genes or are closely associated to the transformation process. The types of aberrations in the G1/S transition seem to be different in various cancers but are nevertheless often linked to clinical behaviors. In this review the role of multiparameter analyses of cell cycle regulatory proteins in breast cancer will be outlined with special attention to pattern analyses as well as the definition of two contrasting pathways in tumorigenesis defined by either cyclin D1 or cyclin F overexpression. PMID- 11883532 TI - Rho GTPases in transformation and metastasis. AB - During the development and progression of human cancer, cells undergo numerous changes in morphology, proliferation, and transcriptional profile. Over the past couple of decades there have been intense efforts to understand the molecular mechanisms involved, and members of the Ras superfamily of small GTPases have emerged as important players. Mutated versions of the Ras genes were first identified in human cancers some 20 years ago, but more recently, the Rho branch of the family has been receiving increased attention. In addition to the experimental evidence implicating Rho GTPase signaling in promoting malignant transformation, genetic analysis of human cancers has now revealed a few examples of direct alterations in the genes encoding regulators of Rho GTPases. In this review, we discuss the evidence implicating Rho GTPases in transformation and metastasis, as well as the progress made toward identifying their biochemical mechanism of action. PMID- 11883533 TI - Transvaginal sonographic cervical length changes during normal pregnancy. AB - OBJECTIVE: To examine the relationship between cervical length and gestational age in normal pregnancy in nulliparous versus parous women. METHODS: We studied a cross-sectional sample of 321 pregnant women, including 185 nulliparous and 136 multiparous women. The inclusion criteria were sonographic confirmation of gestational age within the 12th week, the absence of any risk factors for preterm birth, and uncomplicated pregnancy with expected delivery during the 38th to 42nd weeks. Cervical length was measured in a straight line if the cervix did not show any curvature; in the presence of cervical curvature, the measurement was broken down into 2 or more segments. RESULTS: There was a relationship between gestational age and cervical length, which could be described with a linear function (R = 0.92; R2 = 0.85; P < .001). Moreover, there was no statistically significant difference between multiparous and nulliparous women. CONCLUSIONS: Our study shows that cervical length is comparable in nulliparous and multiparous women throughout pregnancy. In both groups, it actually shows a progressive, linear reduction between the 10th and 40th weeks. Reference ranges constructed for the whole gestational period might be more useful than a single cut-off value for more efficient prevention and management of preterm birth. PMID- 11883534 TI - The added value of transvaginal sonohysterography over transvaginal sonography alone in women with known or suspected leiomyoma. AB - OBJECTIVE: To assess whether sonohysterography provides added diagnostic value over transvaginal sonography in patients with suspected or known myomas by comparing diagnostic confidence, interobserver agreement, accuracy, and change in diagnoses when 2 independent observers interpreted transvaginal sonography alone and later interpreted transvaginal sonography and sonohysterography together. METHODS: Hard copy images from 72 women were interpreted independently by 2 sonologists on separate occasions, rating parameters (abnormal uterus, myoma in any location, submucous myoma, classification of location of a submucous myoma with respect to the uterine cavity, myoma remote from the cavity, adenomyosis, and focal and diffuse endometrial lesions) on a scale of 1 to 5 (1 indicated definitely no; 2, probably no; 3, uncertain; 4, probably yes; and 5, definitely yes). Correlation was made with clinical and imaging follow-up, surgery, and pathologic examination. RESULTS: The added information provided by sonohysterography resulted in improved diagnostic confidence for most parameters. Interobserver agreement was markedly improved for the diagnosis and location of submucous myomas and focal endometrial lesions. Sensitivity values for submucous myomas and focal endometrial lesions were 100% and 90% for transvaginal sonography and sonohysterography together and 100% and 70% for transvaginal sonography alone. CONCLUSIONS: We found that sonohysterography does provide additional information over transvaginal sonography alone and is an important adjunct to transvaginal sonography in symptomatic women with known or suspected myomas, particularly before surgical or medical therapy. PMID- 11883535 TI - Dual-spectra ultrasonography: an attenuation-compensating technique for myocardial perfusion analysis. AB - OBJECTIVE: To validate the harmonic-fundamental frequency ratio peak for user independent differentiation of myocardial perfusion under clinically relevant levels of signal attenuation. METHODS: Radio frequency data were obtained by using apical long-axis scans in 11 open-chest pigs during continuous infusion of a contrast agent after left anterior descending artery occlusion. Silicone pads were interposed between the transducer and the heart to simulate levels of thoracic wall attenuation. Samples of image data from perfused and nonperfused regions were collected; values using harmonic-fundamental frequency ratio peak and conventional harmonic gray scale intensity techniques were calculated. RESULTS: At each attenuation level, the harmonic-fundamental frequency ratio peak value of perfused myocardium was higher than that of nonperfused myocardium (P < .0001). The variance of these values was smaller than that of the gray scale intensity values (P < .0001), with smaller overlap between harmonic-fundamental frequency ratio peak values differentiating perfused and nonperfused regions. In the receiver operating characteristic curves, this analysis had better diagnostic performance than gray scale analysis. In the optimal cutoff value, harmonic fundamental frequency ratio peak analysis provided 87% sensitivity and 91% specificity; gray scale analysis had 80% sensitivity and 78% specificity. CONCLUSIONS: Harmonic-fundamental frequency ratio peak analysis differentiated perfused from nonperfused myocardium under clinically relevant attenuation conditions and provided higher sensitivity and specificity for perfusion determination in attenuated myocardium than did gray scale intensity analysis. PMID- 11883536 TI - Determination of sensitivity versus frequency characteristics of miniature ultrasonic hydrophones below 1 MHz using planar scanning technique. AB - OBJECTIVE: To develop and optimize a rapid and reliable ultrasonic hydrophone calibration procedure suitable for use in the frequency range from 100 to 1000 kHz. METHODS: Planar scanning technique was developed to determine the low frequency response (<1 MHz) of an ultrasonic polyvinylidene difluoride membrane hydrophone probe. RESULTS: The frequency response of a bilaminar membrane design is presented in terms of end-of-cable voltage sensitivity versus frequency from 0.3 to 1 MHz and compared with the sensitivity data determined by other calibration techniques. The experimental data indicate that the sensitivity variation of the hydrophone determined by using the planar scanning technique is approximately +/-1 dB, which agrees well with that obtained using independent calibration methods. CONCLUSIONS: The planar scanning technique is suitable for absolute calibration of hydrophone probes in the frequency range below 1 MHz to within +/-1 dB. Also, the approach developed offers an alternative to other primary calibration techniques such as the reciprocity or broadband pulse technique. The results of this work are important for correctly determining the mechanical index, which is widely accepted as a predictor of potential bioeffects. PMID- 11883537 TI - Postnatal outcome of fetuses with the prenatal diagnosis of gastroschisis. AB - OBJECTIVE: To assess the postnatal outcome and complications that arise in infants with the prenatal diagnosis of gastroschisis. METHODS: Prenatal sonograms with the diagnosis of gastroschisis were identified. Maternal age, indication for sonography, gestational age at diagnosis, other sonographic abnormalities, and postnatal outcome were recorded. RESULTS: Twenty-six fetuses at 16 to 36 weeks' gestational age had gastroschisis diagnosed on sonography. In 5 cases, other fetal anomalies were identified, including hydronephrosis and asymmetric hydrocephalus. In 9 of 21 cases followed by serial prenatal sonography, bowel dilatation developed, prompting delivery in 2. Two of the 26 study fetuses were electively terminated. The remaining 24 were born live and had immediate repair of the gastroschisis after birth. Nineteen infants (79%) had postnatal complications, some with multisystem complications, including 3 deaths, 10 with gastrointestinal complications, 6 with infectious complications, and 6 with anomalies involving other systems (genitourinary, cardiac, central nervous system, and respiratory). Only 5 infants (21%) had completely uncomplicated postsurgical courses. Hospital stays for survivors ranged from 10 to 98 days (mean, 38 days; median, 33 days). CONCLUSIONS: Although reported survival rates are good for gastroschisis, the postoperative hospital stay is often lengthy, and complications are very common, especially those related to the gastrointestinal tract. Other anomalies are uncommon but not rare. PMID- 11883538 TI - Sonographically guided core needle biopsy of bone and soft tissue tumors. AB - OBJECTIVE: To determine the value of sonographically guided core needle biopsies of musculoskeletal tumors as a reliable alternative to fluoroscopy and computed tomography. METHODS: A prospective study was performed in 74 patients referred for image-guided needle biopsy of primary or recurrent musculoskeletal neoplasms and suspected solitary metastasis. Imaging studies performed before biopsy established the feasibility of sonographic guidance in 65 lesions, of which 38 were soft tissue tumors and 27 were bone lesions with extraosseous masses. The lesions were located mainly in the appendicular skeleton. Tissue samples were obtained with a 14-gauge cutting needle coupled to an automated biopsy device under local anesthesia and sonographic guidance. Statistical analysis was based on 48 biopsies confirmed by successful clinical treatment (10 cases) or surgical resection (38 cases). RESULTS: An accurate diagnosis was obtained in 47 (97%) of 48 biopsies; sensitivity was 96%, and specificity was 100%. The method did not yield sufficient tissue to establish a diagnosis in 1 case. Considering all 65 biopsies, high-quality specimens were obtained in 96%. The procedure was carried out expeditiously, and there were no complications. CONCLUSIONS: Sonographically guided core needle biopsy is accurate and safe, obviating open biopsy in most soft tissue masses and bone tumors with extraosseous masses in the appendicular skeleton. In such patients, the sonographically guided procedure is the most prompt and effective method for obtaining tissue samples. PMID- 11883539 TI - Low-intensity pulsed ultrasound enhances early healing of medial collateral ligament injuries in rats. AB - OBJECTIVE: To investigate the effect of low-intensity pulsed ultrasound exposure on the healing of injured medial collateral ligaments. METHODS: Thirteen male Sprague Dawley rats were used in the study. After surgical transection of the bilateral medial collateral ligaments, the ligament of 1 knee received low intensity pulsed ultrasound exposure (30 mW/cm2 for 20 minutes daily), whereas no ultrasound was applied to the contralateral knee (control side). Eight rats were killed at 12 days after surgery, and 5 rats were killed at 21 days. The bilateral knees of 5 rats were used for mechanical testing at each of the 2 periods, and 12 day specimens of the remaining 3 rats were prepared for the electron microscopic examination. The knees of 5 additional rats were used to obtain mechanical data of the normal uninjured medial collateral ligament. RESULTS: On the 12th day, the low-intensity pulsed ultrasound-treated side exhibited significantly superior mechanical properties when compared with the control side in ultimate load, stiffness, and energy absorption (P < .05). However, the treatment did not afford any mechanical advantage when tested on the 21st day. The mean diameter of the fibril was significantly larger on the treatment side than on the control side (P < .05). CONCLUSIONS: Low-intensity pulsed ultrasound exposure is effective for enhancing the early healing of medial collateral ligament injuries. PMID- 11883540 TI - Evaluation of calcific tendonitis of the rotator cuff: role of color Doppler ultrasonography. AB - OBJECTIVE: To use color Doppler ultrasonography to evaluate the morphology and vascularity of calcific tendonitis and to predict the formative and resorptive phases of the calcification. METHODS: Ninety-four patients with shoulder calcification on plain radiographs were enrolled in this study. Ultrasonography of the shoulder was focused on the rotator cuff. Color Doppler ultrasonography was applied in the calcific region. Patient symptoms were graded as painless, mild, moderate, and severe. The calcific plaques were classified as arc-shaped, fragmented or punctate, nodular, and cystic types. Color Doppler ultrasonographic signals were graded 0 to 3. The formative and resorptive phases of calcification were categorized by patient symptoms; acute onset of moderate or severe pain indicated the resorptive phase. RESULTS: The calcific plaques appeared arc shaped in 59 patients (20 painless, 19 mild, and 20 moderate), fragmented or punctate in 27 (2 painless, 3 mild, 20 moderate, and 2 severe), nodular in 6 (1 moderate and 5 severe), and cystic in 2 (severe). There was a significant difference between the morphology of the calcific plaques and clinical symptoms (P < .01). On color Doppler ultrasonography, grade 0 signals were found in 28 patients (22 painless and 6 with mild pain); grade 1 in 18 (16 mild and 2 severe); grade 2 in 41 (all moderate); and grade 3 in 7 (all severe). The correspondence between color Doppler ultrasonographic findings and clinical symptoms was excellent (P < .01). CONCLUSIONS: High-resolution ultrasonography with color Doppler imaging could differentiate the formative and resorptive phases of the calcification and could be used as a follow-up modality in calcific tendonitis of the shoulder. PMID- 11883541 TI - Visualization of uveal perfusion by contrast-enhanced harmonic ultrasonography at a low mechanical index: a pilot animal study. AB - OBJECTIVE: To evaluate contrast-enhanced harmonic ultrasonography at a low mechanical index for its usefulness in visualizing uveal perfusion. METHODS: The study was performed with 9 rabbits, 6 intact and 3 with focal impaired blood flow in the uvea. Ultrasonography was performed by harmonic imaging (transmit, 5 MHz; receive, 10 MHz) with a contrast agent. The agent was administered at a dose of 50 microL/kg. Transmission power was at a mechanical index of 0.2, which is below the US Food and Drug Administration guideline. The images were compared between the impaired and intact eyes. For uveal measurements, video signal intensity versus-time plots were generated in all cases. The plots were analyzed to obtain the rate of signal intensity increase and peak signal intensity. RESULTS: A clear increase of signal intensity was observed after contrast agent administration. The signal intensity of the uvea was lower in the impaired eye than in the intact eye. In the impaired eye, the intensity was lower on the side with impaired flow than on the other side. The differences were significant. CONCLUSIONS: Our findings suggest that uveal perfusion can be visualized by contrast-enhanced harmonic ultrasonography in the harmonic imaging mode at a low mechanical index. PMID- 11883542 TI - Effect of a nitric oxide donor on the ophthalmic artery flow velocity waveform in preeclamptic women. AB - OBJECTIVE: To evaluate the effects of an antihypertensive agent on the orbital circulation of preeclamptic women. METHODS: We studied the ophthalmic arteries of 10 healthy pregnant women and 10 women with severe preeclampsia by pulsed Doppler ultrasonography and evaluated the effect of transdermal isosorbide dinitrate, a nitric oxide donor, on preeclamptic women. RESULTS: The average pulsatility index and resistive index were significantly lower, whereas the average end-diastolic velocity, time-averaged mean peak velocity, and peak ratio, which quantifies characteristic changes in the ophthalmic artery flow velocity waveform, were higher in preeclamptic women. Transdermal isosorbide dinitrate significantly reduced the average end-diastolic velocity (P < .05) and peak ratio of the ophthalmic artery (P < .01), whereas it did not significantly affect other indices. CONCLUSIONS: Orbital circulation was altered in preeclamptic women. A nitric oxide donor affected orbital circulation. Peak ratio was a sensitive index for evaluating orbital circulation in preeclampsia. PMID- 11883543 TI - High-resolution sonography of lower extremity peripheral nerves: anatomic correlation and spectrum of disease. AB - OBJECTIVE: The value of sonography for the diagnosis of diseases of the peripheral nervous system is only little known. This image presentation is intended to raise the awareness of sonographers and clinicians of the potential of sonography by giving an anatomic-sonographic correlation of lower extremity peripheral nerves and an overview of commonly encountered diseases. METHODS: On 2 lower extremity cadaver specimens, peripheral nerves were imaged in typical locations such as the tarsal tunnel. During sonography the nerve was injected with blue dye, and thin-slice anatomic sections were obtained at the scan level with a chain saw. In addition, sonographic images of patients with typical diseases are shown. RESULTS: An excellent anatomic-sonographic correlation was obtained, which underlines the feasibility of sonography for imaging of lower extremity peripheral nerves. Reliable results may be obtained with sonography of typical disease processes of lower extremity peripheral nerves. CONCLUSIONS: Recent developments in sonographic technology such as the introduction of high frequency linear array transducers, compound imaging, and extended field-of-view imaging strongly improve the applicability of transcutaneous sonography for the examination of peripheral nerve disease. PMID- 11883544 TI - Endoluminal sonography of the genitourinary and gastrointestinal tracts. AB - OBJECTIVE: Endoluminal sonography with high-frequency catheter-based transducers is a technique well suited to imaging structures beyond the lumen of the hollow viscus. The purpose of this article was to review some aspects of endoluminal sonography, including instrumentation, clinical applications in the gastrointestinal and genitourinary tracts, and its three-dimensional reconstruction. METHODS: The development of 6F to 10F catheter-based ultrasonic probes has made this technique available for use within a variety of lumina. Endoluminal sonography with frequencies of 9 to 20 MHz has been used for evaluation of a wide range of abnormalities in both the genitourinary and gastrointestinal tracts. RESULTS: Uses in the gastrointestinal tract include quantification of esophageal varices, distinguishing between various submucosal lesions, and measuring the degree of fibrosis in scleroderma. In the genitourinary system, endoluminal sonography has been used to guide collagen injection, to diagnose urethral diverticula and upper tract neoplasms, to locate crossing vessels and septa for guiding endopyelotomy, and to identify submucosal calculi. CONCLUSIONS: High-resolution endoluminal sonography is a new sonographic approach for evaluation of the genitourinary and gastrointestinal tracts. This should lead to the expansion of the diagnostic capabilities of sonography, providing important information for decision making relative to patient care and minimally invasive interventional procedures. Reconstructed three-dimensional endoluminal sonography has the potential to become a valuable tool in both the research and clinical areas. PMID- 11883546 TI - Second-trimester sonographic diagnosis of nonrhizomelic chondrodysplasia punctata. PMID- 11883545 TI - Slipping rib syndrome: a place for sonography in the diagnosis of a frequently overlooked cause of abdominal or low thoracic pain. AB - OBJECTIVE: To describe the sonographic appearance of a poorly recognized cause of low thoracic or upper abdominal pain. METHODS: Three sonographic descriptions of slipping rib syndrome are presented. RESULTS: The 3 patients had abnormal mobility of a cartilaginous rib, which could slip over an adjacent rib during abdominal muscle contraction. CONCLUSIONS: Slipping rib syndrome should be considered in patients with histories of upper abdominal or low thoracic pain of unknown origin. We suggest that high-resolution sonography of the costal margin should be added to abdominal sonography in cases of nonspecific abdominal pain. PMID- 11883547 TI - Prenatal detection of echogenic bowel in a fetus with familial microvillous atrophy. PMID- 11883548 TI - Upper limb phocomelia associated with increased nuchal translucency in a monochorionic twin pregnancy. PMID- 11883550 TI - After-effects of using a weighted bat on subsequent swing velocity and batters' perceptions of swing velocity and heaviness. AB - In baseball and softball, warm-up swings with a weighted bat have been believed to increase swing velocity when an ordinary bat is used in the subsequent competitive situation. The purpose of this study was to investigate the after effects of using a weighted bat on subsequent swing velocity and batters' perceptions of swing velocity and heaviness. Eight men in varsity softball and baseball hit a ball suspended from the ceiling 45 times (3 sets of 15 trials). For each set, the initial 5 trials were done using an ordinary 920-g wooden bat (Control condition), and the following 5 trials by a bat with an 800-g bat ring (Weighted condition), and the final 5 trials again by the ordinary bat (post Weighted condition). Analysis of variance showed a significant decrease of 3.3% in the measured linear velocity of the bat prior to impact with the ball for the first swing of the post-Weighted condition compared with the Control condition. From the second swing the velocity returned to the level of the Control condition. Subjective judgment of the heaviness and velocity of swings for the Weighted and post-Weighted conditions by each participant showed that the ordinary bat felt lighter and swing speed felt faster for the post-Weighted condition. The advantage of the warm-up with a weighted bat was thus psychological and not biomechanical. PMID- 11883549 TI - Effects of single trial of heart-rate biofeedback on the arterial blood pressure, ventilation volume, and oxygen consumption during ramp bicycling exercise. AB - We examined whether the heart rate attenuation resulting from a single trial of biofeedback during exercise occurs without any changes in oxygen consumption (VO2), ventilation volume (V(E)), and mean arterial blood pressure (M(AP)) although these variables are essential determinants for the heart-rate response during exercise. 35 subjects exercised on a cycle ergometer under two conditions while they exercised only (Control) or were trying to decrease the heart-rate response as low as possible by monitoring their own heart-rate responses as Single trial of biofeedback signals. 17 subjects could attenuate their heart rate (Can group), and their heart-rate reduction during Single trial of biofeedback was 5+/-1 bpm. The remaining subjects were unable to reduce heart rate (Cannot group). The heart-rate attenuation during Single trial of biofeedback in the Can group occurred independently of changes in VO2 and MAP but was accompanied by a significant decrease in V(E) and rating of perceived exertion (RPE). However, there was a similar decrease in V(E) in the Cannot group. These findings suggested that the heart-rate attenuation during Single trial of biofeedback was not induced by the metabolic demand of VO2 and the regulations of M(AP) and V(E) during exercise. Other mechanisms, which are probably related to the reduction of RPE, might play an important role in the heart-rate attenuation during a Single trial of biofeedback. PMID- 11883551 TI - Effects of single trial of heart-rate biofeedback during ramp bicycling exercise. AB - We examined whether a single trial of heart-rate biofeedback was effective to attenuate heart-rate responses during ramp exercise despite a lack of biofeedback conditioning. 35 healthy women exercised in two trials while they tried to attenuate their heart rate by watching biofeedback signals or they only exercised without biofeedback signals (Control trial). 17 subjects were able to attenuate the heart rate during Biofeedback trial (Can group) whereas the remaining subjects were not (Cannot group). In the Can group, the magnitude of heart-rate attenuation in all exercise time was equivalent to 11+/-3% of the preexercise heart rate. Since the heart-rate reduction was similar to that achieved after the heart-rate biofeedback conditioning in previous studies, it is likely that the single trial of heart-rate biofeedback was effective for almost half the subjects to attenuate the heart-rate responses during ramp exercise. PMID- 11883552 TI - Effects of pyridoxine on dreaming: a preliminary study. AB - The effect of pyridoxine (Vitamin B-6) on dreaming was investigated in a placebo, double-blind study to examine various claims that Vitamin B-6 increases dream vividness or the ability to recall dreams. 12 college students participated in all three treatment conditions, each of which involved ingesting either 100 mg B 6, 250 mg B-6, or a placebo prior to bedtime for a period of five consecutive days. The treatment conditions were completely counterbalanced and a two-day wash out period occurred between the three five-day treatment blocks. Morning self reports indicated a significant difference in dream-salience scores (this is a composite score containing measures on vividness, bizarreness, emotionality, and color) between the 250-mg condition and placebo over the first three days of each treatment. The data for dream salience suggests that Vitamin B-6 may act by increasing cortical arousal during periods of rapid eve movement (REM) sleep. An hypothesis is presented involving the role of B-6 in the conversion of tryptophan to serotonin. However, this first study needs to be replicated using the same procedures and also demonstrated in a sleep laboratory before the results can be considered certain. PMID- 11883553 TI - Smiling, frowning, and autonomic activity in mildly depressed and nondepressed men in response to emotional imagery of social contexts. AB - The study examined self-reported emotion and facial muscle and autonomic activity of depressed and nondepressed men in response to the social context of emotional situations. 20 university men, assessed on the Beck Depression Inventory, were asked to imagine happy and sad situations with and without visualizing other people. No differences were found between men classified as depressed and nondepressed on self-reported emotion and facial muscle activity. Smiling did not show differences between social contexts although self-reported happiness was increased during happy-social compared to happy-solitary imagery. Adjusting smiling for social context differences in happiness showed less smiling during happy-social than during happy-solitary imagery. In contrast, self-reported sadness and frowning were greater during sad-social compared to sad-solitary imagery. No differences between social contexts were found when frowning was adjusted for social context differences in sadness. Depressed-scoring men showed higher mean heart rate during sad-social than sad-solitary imagery whereas nondepressed-scoring men showed higher mean heart rate during happy social compared to happy-solitary imagery. The results indicate that men may frown more when sad but generally do not smile more during happy-social imagery, independent of depression. Depressed mood may affect heart rate during sad imagery but may not alter facial muscle activity and self:reported emotion in men. PMID- 11883554 TI - Tonguedness in cats. AB - The present study describes a preferential tongue movement in cats, suggesting a so-called "tonguedness" also in animals. 8 cats (66.7%) showed right-tonguedness and 3 left-tonguedness. PMID- 11883555 TI - Predicting performance in ski and swim championships: effectiveness of mood, perceived exertion, and dispositional optimism. AB - Two studies were performed with young athletes to investigate the utility of three psychological tests regarding the prediction of sport performance: the Profile of Mood States. Ratings of Perceived Exertion, and Dispositional Optimism. In Study 1, young male and female cross-country skiers and ski-marksmen in final preparation for the Junior National Swedish Championships were tested. Measured 5 wk. before both competitions, the higher the optimism (LOT), the better performance. In Study 2, which tested young swimmers in preparation for the Senior National Swedish Championships, competitors who had the highest scores on optimism performed less well during the competitions. The results are interpreted to indicate that optimism presents an important factor for predicting achievement in sports. PMID- 11883556 TI - Graphing calculators and students' conceptions of the derivative. AB - This study compared the effect of using graphing calculators on college students' conceptual understanding of the derivative with the traditional teaching approach in Calculus I course. Students (49 men and 52 women) in four classes at a large public university participated. On a posttest, women who were taught calculus using the graphing calculator had significantly higher scores than those taught by the traditional method. PMID- 11883557 TI - Sex-specific finger-length patterns linked to behavioral variables: consistency across various human populations. AB - In humans, as in nonhuman primates, the digits of the hands are similar in length during early fetal development. Subsequently, differentiation leads to a patter of unequal finger lengths, described by George as the finger-length pattern. Recent work by Manning and colleagues suggested that digit length patterns are due to early influences of sex hormones. Most importantly for psychology, such patterns might also relate to cognitive activities that are influenced by early organizing actions of sex hormones. The exciting possibility of having an easily measurable indicator of early action of sex hormones that relates to behavior led us to examine the universality of digit length patterns. With samples from Brazil, Canada, India, Turkey, and Korea, we showed that patterns of distal extent of finger tips are similar across different human populations. Consistent sex differences were found across the samples, showing that the index finger in males extends less far distally relative to the middle finger than is the case for females and that the difference in distal extent between index and ring fingers, relative to the middle finger, is smaller in females than in males. PMID- 11883558 TI - Lack of association of computer use and ability with spontaneous mental visualization. AB - Relationships between frequency of computer use or ability to use the computer effectively with the tendency to construct and process visual mental images were investigated by administering a computer-use questionnaire and a visualization questionnaire to a sample of 185 Italian and Spanish undergraduates. Analysis did not support associations between either (a) frequent computer use of any kind or (b) high competence in using the computer for various purposes and spontaneous use of imagery. PMID- 11883559 TI - Factor structure of an Arabic translation of the scale, preference for numerical information. AB - An Arabic translation of Viswanathan's 1993 scale, Preference for Numerical Information, was administered to 157 tenth-grade students (M age = 16.1 yr.) in the United Arab Emirates. Analysis showed that the scale was homogeneous as the factor solution was comparable to that reported in the original study and item scores were moderately correlated with total scores when used in a different cultural setting. PMID- 11883560 TI - The body-image questionnaire: an extension. AB - The 19-item Body-Image Questionnaire, developed by our team and first published in this journal in 1987 by Bruchon-Schweitzer, was administered to 1,222 male and female French subjects. A principal component analysis of their responses yielded an axis we interpreted as a general Body Satisfaction dimension. The four-factor structure observed in 1987 was not replicated. Body Satisfaction was associated with sex, health, and with current and future emotional adjustment. PMID- 11883561 TI - Relationship between body image and percent body fat among British school children. AB - The aim of the study was to examine the relationship between body image and percent body fat in British school children. A total of 223 11- to 14-yr.-old children from central England took part in the study. Body image was measured using a self-report questionnaire, and percent body fat was measured using skinfold indices. Analysis indicated that body image and adiposity were significantly (p<.01) related for the whole sample, for boys and girls and for White children, Black children, and Asian children. An analysis of variance further indicated significant differences in body image and adiposity between boys and girls, with boys having a more positive body image and a lower percent body fat. Differences (p<.05) were also evident between Black and Asian children, with Black children having a more positive body image and lower percent body fat. No differences were evident between Black and White children or Asian and White children. PMID- 11883562 TI - Female laryngectomees' satisfaction with communication methods and speech language pathology services. AB - 132 female laryngectomees responded to a questionnaire regarding satisfaction with their communication methods as postlaryngectomees and speech-language pathology services. Respondents were satisfied overall with their primary communication selection; however, most would like to try a method that produced a more feminine-sounding voice. 68% were satisfied with their speech-language pathology services. PMID- 11883563 TI - Color and number preferences of patients with psychiatric disorders in eastern Turkey. AB - Color and number preferences have not been studied in healthy sublets and psychiatric patients in Turkey. The study group consisted of a total of 500 patients who had been referred to Firat University Medical Faculty Psychiatry Clinic between March and July 2000 and diagnosed with a psychiatric disorder according to DSM-IV criteria. Preferred colors were requested as a selection from the Luscher Color Test. Then the patients were told to choose a number between 0 and 9. Green was the most frequently preferred of all colors and the number 3 was the most preferred number. Our results demonstrate that patients' choice of color and number reflect the region's religious and cultural milieu. PMID- 11883564 TI - Effect of traditional judo training on aggressiveness among young boys. AB - This study assessed the effect of one year of traditional judo training on aggressiveness among young boys. 27 primary school pupils and 28 judo students were asked to complete the Buss-Perry Aggression Questionnaire at two times 1 year apart. Analysis showed that judoka were more aggressive (had higher scores on Total Aggression, Verbal Aggression, and Anger) than the control group after one year of training, even if variations in aggressiveness were not significant. So, results do not support the view that judo training leads to less aggressiveness in a sample of children this young. PMID- 11883565 TI - Comparison of sport achievement orientation between wheelchair and able-bodied basketball athletes. AB - Differences in sport achievement orientations between 31 recreational wheelchair and 76 able-bodied basketball athletes were tested. Athletes from the New England region completed the three subscales of the Sport Orientation Questionnaire (competitiveness, win orientation, and goal orientation). Wheelchair athletes responded higher on the Competitiveness and Goal Orientation subscales. In discriminative function analysis competitiveness scores were the only significant discriminator between the two groups. PMID- 11883567 TI - Age-related susceptibility to the Muller-Lyer and the horizontal-vertical illusions. AB - Participants between the ages of 3 and 20 years adjusted the Muller-Lyer illusion and the inverted-T form of the Horizontal-Vertical illusion. Perceptual error was quantified using signal detection and nonparametric measures of sensitivity and responsivity. Significant changes in sensitivity and responsivity were found for each illusion across participants' ages. No effect of sex of participant was found. Sensitivity and responsivity were largely asymptotic between the ages of 13 to 15 years for the Muller-Lyer illusion and between the ages of 11 to 15 years for the Horizontal-Vertical illusion. PMID- 11883566 TI - Sleeping pattern of kindergartners and nursery school children: function of daytime nap. AB - With a questionnaire answered by parents this study investigated the sleeping pattern of children attending kindergartens or nursery schools and the function of an afternoon nap. Sleeping pattern was investigated by a questionnaire with 441 young children (229 boys and 212 girls) attending kindergartens or nursery schools at the ages of 3 to 6 years old. Nighttime sleep did not show any significant change, while daytime naps decreased drastically and almost disappeared by the age of 6. Nursery school children went to bed later at night, so nighttime sleep was shorter than that of kindergartners. They also reported having more 'difficulty to fall asleep', more frequent 'staying-up at late night', less 'not getting enough sleep', worse 'mood at rising', and more 'unwillingness to go to their schools'. To investigate whether afternoon naps, which are routine at Japanese nursery schools, can compensate for sleep insufficiency on the previous night and whether they have the effect of delaying the onset of the subsequent nighttime sleep, we compared sleep duration on the previous night and the sleep onset time between the days with and without an afternoon nap. The afternoon nap appeared to cause delayed sleep onset but was not a result of sleep deficiency. PMID- 11883568 TI - Use of an ecologically based program for teaching motor skills in a naturalistic setting to individuals with disabilities. AB - The purpose of this action study was to apply an ecological program for teaching motor skills to individuals with disabilities in naturalistic settings. The program was developed based on an ecological survey. Participants were 22 individuals with disabilities, ages 7 to 26 years in the Fall of 1998 and 18 individuals with disabilities, ages 7 to 24 in the Winter of 1999. In each semester, a participant received one or two 50-min, training session(s) per week for 10 weeks. The number of steps in a motor-activity task performed correctly by a participant was measured in the first and last sessions in each semester. The magnitude of increase in the motor performance by participants and answers to questions in a short survey by participants' parents were used to describe the group performance. Analysis showed group performance of targeted motor skills could be improved using this ecological program in this local learning lab for special physical education. PMID- 11883569 TI - Intrusive effects of implicitly processed information on explicit memory. AB - This study described the interference of implicitly processed information on the memory for explicitly processed information. Participants studied a list of words either auditorily or visually under instructions to remember the words (explicit study). They were then visually presented another word list under instructions which facilitate implicit but not explicit processing. Following a distractor task, memory for the explicit study list was tested with either a visual or auditory recognition task that included new words, words from the explicit study list, and words implicitly processed. Analysis indicated participants both failed to recognize words from the explicit study list and falsely recognized words that were implicitly processed as originating from the explicit study list. However, this effect only occurred when the testing modality was visual, thereby matching the modality for the implicitly processed information, regardless of the modality of the explicit study list. This "modality effect" for explicit memory was interpreted as poor source memory for implicitly processed information and in light of the procedures used. as well as illustrating an example of "remembering causing forgetting." PMID- 11883570 TI - Slow movement execution in event-related potentials (P300). AB - We examined whether slow movement execution has an effect on cognitive and information processing by measuring the P300 component. 8 subjects performed a continuous slow forearm rotational movement using 2 task speeds. Slow (a 30-50% decrease from the subject's Preferred speed) and Very Slow (a 60-80% decrease). The mean coefficient of variation for rotation speed under Very Slow was higher than that under Slow, showing that the subjects found it difficult to perform the Very Slow task smoothly. The EEG score of alpha-1 (8-10 Hz) under Slow Condition was increased significantly more than under the Preferred Condition; however, the increase under Very Slow was small when compared with Preferred. After performing the task. P300 latency under Very Slow increased significantly as compared to that at pretask. Further, P300 amplitude decreased tinder both speed conditions when compared to that at pretask, and a significant decrease was seen under the Slow Condition at Fz, whereas the decrease under the Very Slow Condition was small. These differences indicated that a more complicated neural composition and an increase in subjects' attention might have been involved when the task was performed under the Very Slow Condition. We concluded that slow movement execution may have an influence on cognitive function and may depend on the percentage of decrease from the Preferred speed of the individual. PMID- 11883571 TI - A warning about statistical significance tests performed on large samples of nonindependent observations. AB - When sample observations are not independent, the variance estimate in the denominator of the Student t statistic is altered, inflating the value of the test statistic and resulting in far too many Type I errors. Furthermore, how much the Type I error probability exceeds the nominal significance level is an increasing function of sample size. If N is quite large, in the range of 100 to 200 or larger, small apparently inconsequential correlations that are unknown to a researcher, such as .01 or .02, can have substantial effects and lead to false reports of statistical significance when effect size is zero. PMID- 11883572 TI - Geophysical variables and behavior: XCVII. Increased proportions of the left sided sense of presence induced experimentally by right hemispheric application of specific (frequency-modulated) complex magnetic fields. AB - 12 young men and women who were not aware of the stimulus order were exposed to 8 configurations of weak 1 microTesla) magnetic fields for 5 min. each, applied primarily over the right parietotemporal region. The numbers of sensed presences along the left side, right side, or front/back, as inferred by button presses at the time of the experience, were recorded. There were significantly (eta2= .37) more experiences along the left side than the right side during the presentations of a frequency-modulated (Thomas) pattern with 3-msec. point durations compared to the presentation of its temporally reversed structure or to patterns that were more or less complex. 40% of all left-sided presences occurred during the 5-min. presentation of this specific frequency-modulated pattern. These results suggest that the subjective lateralization of a sensed presence to the left during right hemispheric stimulation by weak magnetic fields is enhanced by the specific temporal structure of the applied field. PMID- 11883573 TI - Ratings of issues in presidential debates of 2000 by students. AB - To assess potential voters' opinions on a variety of issues relevant to the presidential election of 2000, 45 undergraduates completed a questionnaire prior to viewing the third presidential debate between candidates Bush and Gore. Descriptive statistics indicated that, for this sample, economic issues emerged as most important and familiar while Middle-East issues were the least important and least familiar. PMID- 11883574 TI - Comparison of gait patterns between young and elderly women: an examination of coordination. AB - This study investigated intralimb coordination during walking in young and elderly women using the theoretical model of dynamical systems. 20 women, 10 Young (M age=24.6 yr., SD= 3.2 yr.) and 10 Elderly (M age=73.7 yr., SD=4.9 yr.), were videotaped during free speed gait and gait perturbed by an ankle weight. Two parameters, one describing the phasing relationship between segments (mean absolute relative phase) and the other the variability of this relationship (deviation in phase), were calculated from the kinematics. Two-way analysis of variance (age and weight) with repeated measures on weight indicated that during the braking period the weight increased the mean absolute relative phase between the shank and the thigh and decreased it between the foot and the shank. The Elderly women had significant smaller values for the mean absolute relative phase between the shank and the thigh during the braking period. For the same period, deviation in phase increased for the segmental 'relationship between the shank and the thigh. The findings suggest that changes in intralimb coordination take place with asymmetrical weighting and the aging process. These changes are most clearly present during the braking period. PMID- 11883575 TI - Children's face recognition in different contexts: the role of encoding strategies. AB - In this study, the relationship between face recognition and different facial encoding strategies was investigated. Children (6-8 years, N= 134) participated in both a face recognition task and an encoding task. During the recognition task, they saw 7 target faces in an eyewitness context (video) or in a neutral context (static black and white slides) which they later had to recognize from a set of 21 faces. On the encoding task, the same children had to categorize new faces (schematic and photorealistic) into two categories. The construction of the categories allowed participants to encode the faces either analytically (by focusing on a single attribute) or holistically (in terms of overall similarity). The results showed that face recognition was better in the social than in the neutral context. In the neutral context, only holistic encoding was connected to better face recognition. In the social context, children seemed to use not only information about the faces but also information about the persons. PMID- 11883576 TI - Influence of reference frames on asymmetries in Troxler's effect. AB - In 1804, Troxier discovered that, when an observer fixates on a point in central vision and attends to a peripheral stationary stimulus, the peripheral stimulus eventually fades from awareness. This phenomenon is known as Troxler's effect and is allegedly influenced by spatial attention. Asymmetries in Troxler's effect along horizontal and vertical meridian were a recent discovery. However, viewer- and environment-centered reference frames were aligned in prior studies, making it impossible to assess whether asymmetries correspond to viewer- versus environment-centered coordinate systems. This study was undertaken to (a) replicate the asymmetries in the upright condition among 39 participants without health issues and (b) use the asymmetrics to test contrasting predictions made by viewer- and environment-centered coordinate systems when they are decoupled using an experimental head-tilt condition. The horizontal and vertical asymmetries were replicated and consistent with a viewer-centered rather than an environment centered reference frame. PMID- 11883577 TI - Effects of self-assessment on retention in rule-based learning. AB - The main purpose of most educational and training programs isthat the person will acquire some specified knowledge, retain it until a later time when it will be retrieved and employed to make decisions, select and execute actions, etc. Previous research has indicated that there might be a positive relation between an individual's certainty about the correctness of learned responses and how well the material is retained over shorter time-periods. In the present study, 39 men and 38 women learned the names of eight different hand pliers; they were also assessing their certainty about the names of the pliers. After the learning session, the participants returned after either 1, 6, or 12 wk. for a retention and relearning session. Analysis showed that higher certainty was associated positively with retention. The men were more prone than the women to rate themselves as being "Extremely sure" of being correct, even when they, in fact, were wrong. Also, men and women learned the material equally fast, but the men required significantly fewer trials than the women to relearn material. Inclusion of ratings of certainty offers a convenient way of assessing when training has been sufficient and facilitates detection of misinformation (sure-but-wrong answers). PMID- 11883578 TI - Sex differences in susceptibility to the Poggendorff illusion. AB - This study presents experimental results indicating that there are sex differences in the susceptibility to a geometric optical illusion. Participants (57 male and 39 female undergraduate students) performed 3 trials on a test involving the Poggendorff illusion. Analysis indicated that the magnitude of the illusion diminished significantly with each trial and that the percent perceived error was significantly larger for women than for men. This finding is consistent with the numerous studies which have indicated better visuospatial abilities for men than for women. PMID- 11883579 TI - Temporal accuracy of mentally simulated transport movements. AB - Several studies have indicated a clear correspondence in the durations of active versus mentally simulated actions. The present study examined whether this would also be found when a new and unfamiliar task (the pedalo) was given to 15 sports students aged 21 to 27 years, and the range of mental simulation was extended to cover two different modalities. Despite several modifications of experimental procedure, results always showed high correlations between actual and mental durations similar to those reported for walking by Decety, Jeannerod, and Prablanc in 1989. There were also higher scores on absolute error. However, there were no significant differences between mental simulations with open versus closed eyes. It is concluded that the timing of the execution and mental simulation of closed and cyclical movements is interrelated and based on common mechanisms. PMID- 11883580 TI - T-score and raw-score comparisons in detecting brain dysfunction using the booklet category test and the short category test. AB - Often considered one of the more discriminating tests between normal and brain damaged individuals, the Category Test has been described as a complex measure of new problem solving, logical analysis, concept formation, abstract reasoning, and mental efficiency. Since publication of a booklet format by DeFilippis and McCampbell in 1979, researchers have attempted to develop other versions to reduce time required for administration. The present study compared the sensitivity of the Short Category Test, Booklet Format with the Booklet Category Test. Subjects were 22 male veterans seen for comprehensive neuropsychological evaluation and were classified into a brain-damaged or a control group (ns =11) according to independent neurological or neuroradiological evidence. Comparison of performances based on T scores yielded no significant differences between the groups on either format. Analyses of performance based upon suggested raw score cut-offs for the tests, however, yielded a statistically meaningful difference, but caution is in order when using T scores derived from age- or education-based normative data to suggest the presence of brain dysfunction. Raw score cut-offs yielded valid differentiation of individual patients in both groups. PMID- 11883581 TI - Amount of newspaper coverage of high school athletics for boys and girls on sports page and newspaper circulation. AB - This study analyzed the amount of coverage for high school athletics in 43 newspapers with small circulation by devoting 40% of their interscholastic athletics coverage to girls in athletics, printed significantly more articles about girls' athletics than did the newspapers with medium (33%) or large (32%) circulation. Therefore, the smaller the newspaper circulation, the more equitable the coverage of athletics for girls and boys. This finding was consistent with some prior work but not all. PMID- 11883582 TI - Recognition of items added to or deleted from scenes: method of alterations, type of alterations, and salience. AB - The present study investigated which variable correlated with recognition the most of three variables (method of alteration: Introduction or Restoration, type of alteration: Addition or Deletion, and Salience). Stimuli were pictures of scenes. An item was deleted from an original photograph (deletion change) or the deleted item was restored (addition change). Participants in the Introduction condition viewed original pictures in the learning phase and recognized changed pictures (additions or deletions) in the test phase, and participants in the Restoration condition viewed changed pictures in the learning phase and recognized original pictures in the test phase. Other participants rated salience of the changed items. Analysis showed that an effect of method of alteration was observed only for Deletion and an effect of type of alteration was observed only for Introduction. That we found method of alteration held for pictures of cats, geometric figures, and scenes (present study) shows the robustness of the effects of the method of alteration. The reason of this robustness requires further investigation. PMID- 11883583 TI - Object-recognition tasks: comparing paper versions to computerized laboratory methods. AB - This study attempts to generalize Biederman's 1987 findings regarding Recognition by Components theory, which were obtained using a computer administered object recognition task, to an analog or paper task that is consistent with typical assessment or testing procedures. Three versions of an object-recognition task were developed after the Structure of Intellect-Learning Abilities Test by Meeker, Meeker, and Roid. One version contained randomly fragmented objects, one contained objects with vertices present, and the third contained objects with midsegments. 30 participants were administered each of the three versions in a counterbalanced order. The results are consistent with those of Biederman. Objects with missing vertices were more difficult to recognize than objects with missing midsegments. There was no difference between randomly fragmented objects and those with vertices present. Implications for object-recognition research and test-item development are discussed. In particular, it is suggested that perceptual theories should be used in developing test items to gain greater control in creating items of appropriate difficulty and to increase the validity of the overall instrument. PMID- 11883584 TI - The anxiolytic effect of aqua aerobics in elderly women. AB - This study examined the anxiolytic (anxiety reducing) effects of exercise for elderly women engaging in a single bout of aqua aerobics. Volunteers (N=29) completed questionnaires immediately before and after participating in an aqua aerobics class. The average age of participants was 66.4 yr. A brief form of Spielberger's State Anxiety Inventory and a questionnaire on demographic items were administered prior to engagement in exercise, and the brief form of the State Anxiety Inventory was administered again immediately after the exercise session. There was a significant difference on a t test between participants' ratings of anxiety before exercise (M = 16.8) compared to after exercise (M= 13.9); participants' ratings of state anxiety were somewhat lower after exercising. Weaknesses of the present study and suggestions for research are presented. PMID- 11883585 TI - Confirmatory factor analysis of the group environment questionnaire with co acting sports. AB - To assess whether the Group Environment Questionnaire is applicable to athletes in co-acting sports such as track and field, the factor structure of the Group Environment Questionnaire among a heterogeneous sample of 199 co-acting team sport athletes was investigated. A confirmatory factor analysis did not support the hypothesized four-factor model. A subsequent exploratory factor analysis yielded two independent factors that had no interpretable pattern of the four Group Environment Questionnaire factors within them. While this finding requires replication and confirmation, the four-factor model of the Group Environment Questionnaire does not appear to be as valid for co-acting sports as for interacting sports. PMID- 11883586 TI - Color naming by boys and girls. AB - The present study describes a field experiment conducted to test the hypothesis that color naming varies for 56 adolescent boys and 39 girls. No standard color chart such as Munsell, Pantone, or Crayola was used. Instead, subjects identified 15 computer-generated color samples assigned a score based on thescale devised by Rich in 1977 of four color descriptor categories. Findings differed from those of most previous studies, in which women aged over 16 years showed a more elaborate color vocabulary than men. Among the 14- to 16-yr.-old subjects, there was no significant difference in color naming. PMID- 11883587 TI - Color of computer display frame in work performance, mood, and physiological response. AB - The effects of the color of a personal computer screen on work performance, psychological mood, and autonomic response were investigated. 24 subjects were asked to perform visual tasks presented on the computer display. Three types of computer monitor, which were colored red, blue, or beige, were employed to present visual cognitive tasks. The mood measure, the Japanese Stress Arousal Check List, and heart rate measurement were administered before and after work on each color of computer monitor. Analysis of a low-demand task (Exp. 1) showed that the red computer monitor reduced visual task performance compared to that with the blue, while the blue monitor decreased visual task performance on a high demand task (Exp. 2). The color of the monitor did not affect mood or heart rate. Based on these findings, the effect of the color of environmental cues on work was discussed. PMID- 11883588 TI - Anthropometric profiles and social physique anxiety of physical education professionals from India. AB - Previous work indicates that there may be a relationship between the observation and evaluation of one's physique and the construct of social physique anxiety associated with this process. Since physical educators are expected to serve as role models of desired fitness behaviors, bodily appearance, and composition, it is of interest to examine whether such responses and relationships may be observed. However, there is limited published information on anthropometric profiles and body images of physical education professionals, especially of those from India. Therefore, this study compared anthropometric profiles and Social Physique Anxiety in a sample of 182 male physical education professionals from India (M age=41.2 yr.). Body Mass Index, sum of three skinfolds (Tricep, Abdomen, and Thigh), and waist/hip ratio were determined using standard procedures. The sample was grouped into overweight and normal weight categories. Significant group differences were found for the sum of skinfolds and waist/hip ratio, with no significant differences between groups on the Social Physique Anxiety total score. Correlations for the anthropometric measures with the Social Physique Anxiety scores indicated no significant relationships. Mean total Social Physique Anxiety score for the combined group was comparable to those reported for other groups of physically active individuals. These findings indicate low Social Physique Anxiety in this sample and may have implications with regard to the attitudes pertaining to body image and role modeling of appropriate fitness behaviors. PMID- 11883589 TI - Attitudes of undergraduate majors in elementary education toward mathematics through a hands-on manipulative approach. AB - Using a manipulative approach in a mathematics course, this study was designed to assess attitudes toward mathematics of undergraduates in elementary education. A 20-item attitude rating administered as pre- and posttest to 95 students (7 men and 88 women) was used to assess the effect of the hands-on approach. A dependent t test indicated a statistically significant but very small change in their ratings. PMID- 11883590 TI - Acquisition of expertise on a difficult perceptual-motor task by an amnesic patient. AB - While numerous studies have reported learning of perceptual-motor skills by amnesic patients, few if any have documented the eventual acquisition of expertise on a given task. This paper recounts the learning of the computer game Tetris by a hippocampal amnesic, whose acquisition of the task in a formal evaluation was somewhat slower than that of a comparison group, but who after many hours of self-paced practice achieved expert-level play. PMID- 11883591 TI - Influence of physical exercise on perceptual response in aerobically trained subjects. AB - A significant effect of fatigue induced byphysical exercise leading to exhaustion was observed for 6 male triathletes using some specific analysis of the critical flicker fusion test. PMID- 11883592 TI - Object-related knowledge and the production of gestures with imagined objects by preschool children. AB - Previous research suggests that the gestural representation of preschool age children has a symbolic quality in the absence of real objects. It has shown a developmental progression from use of concrete body parts to more abstract imaginary-object gestural representations during the preschool years. The present study examined whether object-related knowledge is involved in the production of imaginary-object gestures. 35 children (12 3-, 11 4-, and 12 5-yr.-olds) performed gestural tasks in which they were asked to pretend to use common objects, e.g., "pretend to brush your teeth with a toothbrush." In addition, they were asked to describe the relationship between the performed gesture and object when they were pretending. Analysis indicated that, when an imaginary-object gesture was performed, it was through an evoked prior knowledge of the function and context of the use of objects. This may suggest that object-related knowledge is a key factor involved in developmental changes from body part to imaginary object gestures and that it is involved in freeing preschool children from object substitution, which is prescribed by perceptual support. PMID- 11883593 TI - Age, sex, and body mass index in performance of selected locomotor and fitness tasks by children in grades K-2. AB - The purpose of this study was to examine the contributions of 3 predictor variables (age, sex, and body mass index) to performance of 7 fundamental movement skills (locomotor; run, gallop, hop, leap, jump, skip, and slide) and 4 fitness tasks (grip strength, step test, sit and reach, and timed sit-ups) by 65 children in Grades K-2 (M=6 yr.). A multiple regression analysis indicated that running, leaping, and skipping improved with age. No sex differences were observed on any of the seven skills. In terms of the four fitness tasks, age was positively associated with timed sit-ups and grip strength and inversely associated with step-test performance. Sex was associated with step-test and sit and reach performance (girls performed better on both). Finally, body mass index was related to increased grip strength and fewer timed sit-ups. PMID- 11883594 TI - Pseudofluency in adults who stutter: the illusory outcome of therapy. AB - The majority of therapy programs for people who stutter are aimed at modifying the entire speech output by using techniques that reduce the overt signature events. Use of these techniques for extended time periods are thought to induce true fluency that is automatic, natural, and effortless. It is proposed that the present form of therapeutic intervention induces pseudofluency rather than true fluency. Pseudofluency is the speech posrtherapy of persons who stutter, free of the discrete signature events of stuttering, but replaced by cognitively mediated gestures that are embedded as continuous prolongation or masked stuttering events throughout the speech act. This may account for the high rate of relapse and the problems associated with the maintenance, stability, and naturalness of speech after stuttering therapy. PMID- 11883595 TI - Verbal versus olfactory cues: affect in elicited memories. AB - Verbal processing has a reduced role for olfactory stimuli. It is difficult to provide a label for an odor experience. Odor perception can retrieve memories of life events with personal meaning and elicit affective experiences. Odors that have emotionally loaded content could produce older memories. Common odors with well-known names have been used. In Exp. 1 the names were shown, and the subjects were asked to imagine the corresponding odors; subsequently those odorants were presented. In Exp. 2 at first the odorants were presented and subsequently their names, printed one each per white card. The subjects were requested to provide written free associations. At the end of each session they scored a semantic differential. The hypothesis that emotionally loaded associations are more frequent when evoked by odorants seems confirmed, supported also by some reliable differences between the profiles for olfactory verbal stimuli. The evaluation of olfactory stimuli did not differ from one experiment to the other; verbal stimuli, on the contrary, are differently evaluated if the corresponding odorants were presented before or after their labels. PMID- 11883596 TI - Courtship communication and perception. AB - Although ethologists have detailed courtship rituals for many species, courting behavior of humans has not received extensive study from an ethological standpoint. Yet there are clearly facial expressions and gestures that are commonly labeled "flirting behaviors." These nonverbal signals have been documented recently by several investigators in field studies, but the receptivity of nonverbal courtship signals is still in question. The current research project attempted to assess the perceptual skill of naive male and female observers who were presented videotaped samples of females' nonverbal courtship and rejection behaviors and asked to rate their intensity. The results showed that overall, men rated invitational behaviors more positively than women. In contrast, signals of rejection were seen by men as sending a less potent message than that perceived by women. Evolutionary theory may offer a framework for understanding these results. PMID- 11883597 TI - Photophysical and photochemical studies of 2-phenylbenzimidazole and UVB sunscreen 2-phenylbenzimidazole-5-sulfonic acid. AB - The sunscreen agent 2-phenylbenzimidazole-5-sulfonic acid (PBSA) and its parent 2 phenylbenzimidazole (PBI) cause DNA photodamage via both Type-I and Type-II mechanisms when UVB irradiated. We have studied the photophysical and photochemical properties of these compounds and their ability to photogenerate reactive oxygen species including free radicals. PBI and PBSA exhibit both oxidizing and reducing properties in their excited state. The absorption and fluorescence properties of PBSA depend strongly upon pH, and hence the photochemistry of PBSA was studied in both neutral and alkaline solutions. PBSA showed strong oxidizing properties when UV irradiated in neutral aqueous solution (pH 7.4) in the presence of cysteine, glutathione and azide, as evidenced by the detection of the corresponding S-cysteinyl, glutathiyl and azidyl radicals with the aid of the spin trap, 5,5-dimethyl-1-pyrroline N-oxide (DMPO). However, when an aqueous anaerobic solution (pH 10) of PBSA and either nitromethane (NM) or 4 nitrobenzoic acid (4-NBA) were irradiated, the corresponding nitro anion radicals were observed. This finding suggests that both NM and 4-NBA are reduced by direct electron transfer from the excited state PBSA. During UV irradiation of an aerobic solution of PBSA, O2*- and *OH radical were generated and trapped by DMPO. Further, PBI (in ethanol) and PBSA (in ethylene glycol : water 2: 1 mixture) showed low temperature (77 K) phosphorescence (lambdamax = 443, 476 and 509 nm) and also an electron paramagnetic resonance half-field transition (deltaMs = +/-2), which is evidence for a triplet state. This triplet produced singlet oxygen (1O2) with quantum yields 0.07 and 0.04 in MeCN for PBI and PBSA, respectively. These studies demonstrate that UV irradiation of PBSA and PBI generates a variety of free radicals and active oxygen species that may be involved in the photodamage of DNA. PMID- 11883598 TI - Sensitization of lanthanides by nonnatural amino acids. AB - The sensitization of Eu(III) and Tb(III) by ethylenediaminetetraaceticacid (EDTA) derivatized tryptophan (Trp), 7-azatryptophan (7AW) and 5-hydroxytryptophan (5HW) has been examined. These Trp analogs were utilized in the present study because they can be incorporated into proteins in place of native Trp residues and because they absorb strongly beyond 305 nm (where Trp absorbance goes to zero), allowing selective excitation of such species in the presence of other Trp containing proteins. All three indole derivatives were able to sensitize Tb(III) luminescence, with the relative sensitization being in the order Trp > 5HW > 7AW. On the other hand, only the 7AW-EDTA complex was able to sensitize Eu(III) luminescence, likely owing to a better spectral overlap between 7AW emission and Eu(III) absorbance. The sensitized emission of Tb(III) and Eu(II) displayed the expected long emission lifetimes at 545 nm [for Tb(III)] and 617 nm [for Eu(III)], indicating that long-lifetime lanthanide emission could be produced using nonnatural amino-acid donors. Thus, 7AW- and 5HW-sensitized lanthanide emissions should prove to be useful in biophysical studies, such as the use of fluorescence energy transfer to probe biomolecular interactions in vivo. PMID- 11883599 TI - The effect of UV absorbing sunscreens on the reflectance and the consequent protection of skin. AB - The in vivo reflectance spectra of Caucasian skin, coated with preparations containing sunscreen vehicle, vehicle with olive oil and vehicle with the UVB and UVA absorbers 2-ethylhexyl-4-methoxycinnamate and 4-t-butyl-4' methoxydibenzoylmethane were determined. All preparations reduced the reflectance of skin throughout the UVA spectral range (320 to 400 nm), with the sunscreen preparations containing the UVB and UVB plus UVA absorbers reducing the reflectance more than the sunscreen vehicle alone. This phenomenon, which facilitates the penetration of UV radiation to the lower epidermis and dermal layers of skin and therefore lessens sunscreen efficacy, is attributed to optical coupling mediated by refractive index matching of the sunscreen to the upper epidermis. The greater reduction in skin diffuse reflectance caused by sunscreens containing methoxycinnamate is associated with this compound's high refractive index. Also, by determining the excitation spectra of the autofluorescence originating from the dermal layer of skin, the transmission spectra of the various components of sunscreen on skin were established, and these were in good general agreement with previously published spectra. PMID- 11883600 TI - Proton uptake of rhodobacter capsulatus reaction center mutants modified in the primary quinone environment. AB - Flash-induced absorbance spectroscopy was used to analyze the proton uptake and electron transfer properties of photosynthetic reaction centers (RC) of Rhodobacter capsulatus that have been genetically modified near the primary quinone electron acceptor (Q(A)). M246Ala and M247Ala, which are symmetry-related to the positions of two acidic groups, L212Glu and L213Asp, in the secondary quinone electron acceptor (QB) protein environment, have been mutated to Glu and Asp, respectively. The pH dependence of the stoichiometry of proton uptake upon formation of the P+Q(A)- (H+/P+Q(A)-) and PQ(A) (H+/Q(A)-) (P is the primary electron donor, a noncovalently linked bacteriochlorophyll dimer) states have been measured in the M246Ala --> Glu and the M247Ala --> Asp mutant RC, in the M246Ala-M247Ala --> Glu-Asp double mutant and in the wild type (WT). Our results show that the introduction of an acidic group (Glu or Asp) in the QA protein region induces notable additional proton uptake over a large pH region (approximately 6-9), which reflects a delocalized response of the protein to the formation of Q(A)-. This may indicate the existence of a widely spread proton reservoir in the cytoplasmic region of the protein. Interestingly, the pH titration curves of the proton release caused by the formation of P+ (H+/P+: difference between H+/P+Q(A)- and H+/PQ(A)- curves) are nearly superimposable in the WT and the M246Ala --> Glu mutant RC, but substantial additional proton release is detected between pH 7 and 9 in the M247Ala --> Asp mutant RC. This effect can be accounted for by an increased proton release by the P+ environment in the M247Ala --> Asp mutant. The M247Ala --> Asp mutation reveals the existence of an energetic and conformational coupling between donor and acceptor sides of the RC at a distance of nearly 30A. PMID- 11883601 TI - Enhancement of lysosomal osmotic sensitivity induced by the photooxidation of membrane thiol groups. AB - The osmotic lysis of photodamaged lysosomes is a critical event for killing tumor cells. How the photodamage increases lysosomal osmotic sensitivity is still unclear. In this work, the effect of the photooxidation of membrane thiol groups on the lysosomal osmotic sensitivity was studied by measuring the thiol groups with 5,5'-dithiobis(2-nitrobenzoic acid) and examining the lysosomal beta hexosaminidase latency loss in a hypotonic sucrose medium. The results show that methylene blue-mediated photooxidation of lysosomes decreased their membrane thiol groups and produced cross-linkage of membrane proteins (molecular weight ranging from 75000 to 125000), which was visualized by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Simultaneously, the lysosomal osmotic sensitivity increased. These photoinduced alterations of the lysosomes could be recovered by reducing the oxidized thiol groups with dithiothreitol. It indicates that the photooxidation of membrane thiol groups can increase the lysosomal osmotic sensitivity and therefore provides a new explanation for the photoinduced lysosomal lysis. PMID- 11883602 TI - In vitro and in vivo efficacy of photofrin and pheophorbide a, a bacteriochlorin, in photodynamic therapy of colonic cancer cells. AB - This study was designed to investigate the efficacy of photodynamic therapy (PDT) in treating colonic cancer in a preclinical study. Photofrin, a porphyrin mixture, and pheophorbide a (Ph a), a bacteriochlorin, were tested on HT29 human colonic tumor cells in culture and xenografted into athymic mice. Their pharmacokinetics were investigated in vitro, and the PDT efficacy at increasing concentrations was determined with proliferative, cytotoxic and apoptotic assessments. The in vivo distribution and pharmacokinetics of these dyes (30 mg/kg, intraperitoneal) were investigated on HT29 tumor-bearing nude mice. The inhibition of tumor growth after a single 100 J/cm2 PDT session was measured by the changes in tumor volume and by histological analysis of tumor necrosis. PDT inhibited HT29 cell growth in culture. The cell photodamage occurred since the time the concentrations of Ph a and Photofrin reached 5.10(-7) M (or 0.3 microg/mL) and 10 microg/mL, respectively. A photosensitizer dose-dependent DNA fragmentation was observed linked to a cleavage of poly(ADP-ribose) polymerase and associated with an increased expression of mutant-type p53 protein. PDT induced a 3-week delay in tumor growth in vivo. The tumor injury was corroborated by histological observation of necrosis 48 h after treatment, with a correlated loss of specific enzyme expression in most of the tumor cells. In conclusion, PDT has the ability to destroy human colonic tumor cells in vitro and in vivo. This tumoricidal effect is likely associated with a p53-independent apoptosis, as HT29 cells express only mutated p53. The current study suggests a preferential use of Photofrin in PDT of colonic cancer because it should be more effective in vivo than Ph a as a consequence of better tumor uptake. PMID- 11883603 TI - Laser targeted photo-occlusion of rat choroidal neovascularization without collateral damage. AB - Laser targeted photo-occlusion (LTO) is a novel method being developed to treat choroidal neovascular membranes (CNV) in age-related and other macular degenerations. A photosensitive agent, encapsulated in heat-sensitive liposomes, is administered intravenously. A low power laser warms the targeted tissue and releases a bolus of photosensitizer. The photosensitizer is activated after it clears from the normal choriocapillaris but not from the CNV. Forty-five experimental CNV were induced in seven rats. Five weeks after LTO, complete occlusion was observed by laser targeted angiography (LTA) in 76% of treated CNV, and partial occlusion was found in the remaining 24%. The tissues outside the CNV but within the area treated by LTO showed no flow alteration and no dye leakage. All untreated CNV were patent on LTA at 5 weeks. Light microscopy and electron microscopy confirmed the results in treated and control lesions. Moreover, treated areas next to lesions showed normal photoreceptors, retinal pigment epithelium (RPE), Bruch's membrane and choriocapillaris. These results indicate that LTO may improve current photodynamic therapy by alleviating the need for repeated treatments and by avoiding the long-term risks associated with damage to the RPE and occlusion of normal choriocapillaries. PMID- 11883604 TI - Mitochondrial alterations in fanconi anemia fibroblasts following ultraviolet A or psoralen photoactivation. AB - The genetic disease Fanconi anemia (FA), generally considered to be a DNA repair defect, has also been related to a deficiency in cellular defense against reactive oxygen species (ROS). Results show that mitochondrial matrix densification occurs rapidly and transiently in FA fibroblasts following 8 methoxypsoralen (8-MOP) photoreaction or ultraviolet A (320 to 380 nm) (UVA) irradiation. This effect is oxygen dependent because it is more important under 20 than under 5% oxygen tension. In contrast, in normal fibroblasts very little, if any, densification of mitochondrial matrix is induced by treatments even at the highest oxygen tension. The changes in matrix density in FA cells are accompanied by some modifications in transmembrane potential, linked to a Fenton like reaction, and in mitochondrial cardiolipin content, differing from the responses of normal cells. These data are indicative of some sort of membrane damage induced by 8-MOP photoreaction and UVA irradiation, to which FA cells appear to be particularly sensitive. PMID- 11883605 TI - Identification of a light-regulated protein kinase activity from seedlings of Arabidopsis thaliana. AB - Protein kinase transduction pathways are thought to be involved in light signaling in plants, but other than the photoreceptors, no protein kinase activity has been shown to be light-regulated in vivo. Using an in-gel protein kinase assay technique with histone H III SS as an exogenous substrate, we identified a light-regulated protein kinase activity with an apparent molecular weight ca 50 kDa. The kinase activity increased transiently after irradiation of dark-grown seedlings with continuous far red light (FR) and blue light (B) and decreased after irradiation with red light (R). The maximal activation was achieved after 30 min to 1 h with FR or B. After irradiation times longer than 2 h, the kinase activity decreased to below the sensitivity level of the assay. In Arabidopsis mutants lacking either the photoreceptors phytochrome A, phytochrome B or the blue-light receptor cryptochrome 1, kinase activity was undetectable, whereas in the photomorphogenic mutants cop1 and det1 the kinase activity was also observed in the absence of light signals, though still stimulated by B and FR. Interestingly, the R inhibition of the kinase activity was lost in the mutant hy5. Pretreatment with cycloheximide blocked the kinase activity. PMID- 11883606 TI - The role of A2E in prevention or enhancement of light damage in human retinal pigment epithelial cells. AB - The process of sight (photostasis) produces, as a by-product, a chromophore called 2-[2,6-dimethyl-8-(2,6,6-trimethyl-1-cyclohexen-1-yl)-1E,3E, 5E,7E octatetraenyl]-1-(2-hydroxyethyl)-4-[4-methyl-6-(2,6,6-trimethyl-1-cyclohexen-1 yl)-1E, 3E, 5E-hexatrienyl]-pyridinium (A2E), whose function in the eye has not been defined as yet. In youth and adulthood, A2E is removed from human retinal pigment epithelial (h-RPE) cells as it is made, and so it is present in very low concentrations, but with advanced age, it accumulates to concentrations reaching 20 microM. In the present study we have used photophysical techniques and in vitro cellular measurements to explore the role of A2E in h-RPE cells. We have found that A2E has both pro- and antioxidant properties. It generated singlet oxygen (phiso = 0.004) much less efficiently than its precursor trans-retinal (phiso = 0.24). It also quenched singlet oxygen at a rate (10(8) M(-1) s(-1)) equivalent to two other endogenous quenchers of reactive oxygen species in the eye: alpha-tocopherol (vitamin E) and ascorbic acid (vitamin C). The endogenous singlet oxygen quencher lutein, whose quenching rate is two orders of magnitude greater than that of A2E, completely prevented light damage in vitro, suggesting that singlet oxygen does indeed play a role in light-induced damage to aged human retinas. We have used multiphoton confocal microscopy and the comet assay to measure the toxic, phototoxic and protective capacity of A2E in h-RPE cells. At 1 5 microM, A2E protected these cells from UV-induced breaks in DNA; at 20 microM, A2E no longer exerted this protective effect. These results imply that the role of A2E is not simple and may change over the course of a lifetime. A2E itself may play a protective role in the young eye but a toxic role in older eyes. PMID- 11883607 TI - Chlorella virus pyrimidine dimer glycosylase excises ultraviolet radiation- and hydroxyl radical-induced products 4,6-diamino-5-formamidopyrimidine and 2,6 diamino-4-hydroxy-5-formamidopyrimidine from DNA. AB - A DNA glycosylase specific for UV radiation-induced pyrimidine dimers has been identified from the Chlorella virus Paramecium Bursaria Chlorella virus-1. This enzyme (Chlorella virus pyrimidine dimer glycosylase [cv-pdg]) exhibits a 41% amino acid identity with endonuclease V from bacteriophage T4 (T4 pyrimidine dimer glycosylase [T4-pdg]), which is also specific for pyrimidine dimers. However, cv-pdg possesses a higher catalytic efficiency and broader substrate specificity than T4-pdg. The latter excises 4,6-diamino-5-formamidopyrimidine (FapyAde), a UV radiation- and hydroxyl radical-induced monomeric product of adenine in DNA. Using gas chromatography-isotope-dilution mass spectrometry and y irradiated DNA, we show in this work that cv-pdg also displays a catalytic activity for excision of FapyAde and, in addition, it excises 2,6-diamino-4 hydroxy-5-formamidopyrimidine (FapyGua). Kinetic data show that FapyAde is a better substrate for cv-pdg than FapyGua. On the other hand, cv-pdg possesses a greater efficiency for the extension of FapyAde than T4-pdg. These two enzymes exhibit different substrate specificities despite substantial structural similarities. PMID- 11883608 TI - Inhibiting effect of beta-cyclodextrin on thymine dimerization photosensitized by para-aminobenzoic acid. AB - Beta-cyclodextrin can act as an efficient inhibitor of the photosensitized dimerization of thymine by para-aminobenzoic acid (PABA) in aqueous solution. This can be explained by considering the formation of an inclusion complex between PABA and beta-cyclodextrin. PMID- 11883609 TI - Photobiological properties of hydroxy-substituted flavothiones. AB - Flavothione (FT) and a series of 18 hydroxy- and methoxy-substituted flavothiones were screened for photobiological activity. The 5-hydroxy-substituted compounds (group 3) and the methoxy-substituted flavothiones were inactive. FT and the remaining hydroxy-substituted compounds, all displayed photobiological activity. Among these, the 3-hydroxy-substituted compounds (group 2) were the most efficient photosensitizers overall in spite of their concurrent fast photodegradation. FT and all other hydroxyflavothiones, not substituted in the 3- or 5-positions (group 1), were inefficient compared with group 2. Detailed photobiological tests were carried out for four flavothiones of groups 1 and 2. The biological tests included fungi, several strains of Escherichia coli, Salmonella typhimurium and mammalian cells. In addition, the ability of these flavothiones to perform lipid peroxidation was evaluated. FT and 6 hydroxyflavothione (group 1) induce DNA damage via H-atom abstraction from the lowest n, pi* triplet state of the thione (oxygen independent). For 3-hydroxy and 3,6-dihydroxyflavothione (group 2), both DNA and the membrane are targets. The mechanism likely involves both energy transfer and electron transfer from the lowest pi, pi* triplet state to oxygen, to form singlet oxygen and the superoxide anion. Some of these compounds could be considered as models for environmentally safe photopesticides. PMID- 11883610 TI - Legal guidelines related to end-of-life decisions: are nurse practitioners knowledgeable? AB - Public demand and professional standards dictate that primary care providers must be prepared to offer guidance and advance care planning for end-of-life decision making to their patients. The purpose of this study was to describe the relationship between nurse practitioners' (NPs) knowledge of legal guidelines for end-of-life decision-making, their knowledge of the clinical application of advance care planning (ACP), and their comfort in counseling patients on these issues and personal attitudes toward end-of-life care. This descriptive, correlational study used survey data from a convenience sample of 145 Washington State NPs. Most NPs in this study were reasonably knowledgeable about the legal guidelines for end-of-life decision-making and clinical applications for ACP. They were somewhat comfortable with counseling patients on end-of-life decisions and expressed positive attitudes toward end-of-life care. However, a significant number of NPs were ill-informed about the legal guidelines and few actually incorporated ACP into their clinical practice. The authors suggest educational programs should focus on addressing the problem of stable misinformation related to legal guidelines. In addition, education should offer NPs didactic information and role modeling to skillfully incorporate ACP into clinical practice. PMID- 11883611 TI - Special challenges of withholding artificial nutrition and hydration. AB - Despite society's increasing understanding of and comfort with end-of-life decision-making, questions about the appropriate use of artificial hydration and nutrition remain particularly challenging to both professional and family members. These decisions are complicated by misunderstandings about likely benefits and burdens, concern about patient suffering, and ambivalence regarding the moral status of feeding. Data regarding the effectiveness of tube feedings in terms of prolonging survival, improving wound healing, or preventing aspiration do not support the widespread use of this intervention in states of severe dementia or end-stage disease. Further, there is evidence that withholding feeding is not associated with suffering, so long as adequate mouth care and desired sips of water are provided. Nonetheless, surveys of both long-term care residents and family members of persons with dementia indicate that at least 30% to 50% of those queried expresses a preference for artificial feeding if they (or their relative) could not eat. Given the absence of conclusive data about the efficacy of feeding and the apparent plurality of values surrounding the provision of this intervention, health professional must focus efforts on using a careful, deliberate approach to decision-making that involves all interested parties and make use of valid empirical data. PMID- 11883612 TI - Comfort and pain relief in dementia: awakening a new beneficence. PMID- 11883613 TI - Collaboration: a tool addressing ethical issues for elderly patients near the end of life in intensive care units. PMID- 11883614 TI - A case study of the death of an older woman in a nursing home: are nursing care practices in compliance with ethical guidelines? AB - This article presents a case study describing and analyzing ethical issues in the care during the last 11 weeks of life of a 101-year-old nursing home resident. The case presented here is part of a larger ongoing ethnographic study of death and dying in nursing homes. Two nursing care issues with ethical implications are discussed. First, the resident could not eat or drink independently, but she received no assistance with her meals. Second, she remained in a wheelchair for many hours and developed three pressure ulcers on her buttocks. Furthermore, she became tired when sitting in the wheelchair for long periods of time, fell out of the wheelchair, and subsequently was restrained "for her safety." Using selected principles from the International Council of Nurses and the American Nurses Association code for nurses, the resident's care is discussed and recommendations for improving the care of nursing home residents are presented. PMID- 11883615 TI - Nursing care at end-of-life. PMID- 11883616 TI - Facilitating end-of-life decision-making: strategies for communicating and assessing. AB - End-of-life decision-making is often a difficult process and one that many elderly patients and their families will undergo. The grounded theory study of nurses, physicians, and family members (n = 20) reported in this article examined provider behaviors that facilitated the process of decision-making near the end of patients' lives. According to participants, providers who are experienced and comfortable are more likely to engage in communication and assessment strategies that facilitate end-of-life decision-making. Communication strategies included: being clear, avoiding euphemisms, spelling out the goals and expectations of treatment, using words such as "death" and "dying," and being specific when using such words as "hope" and "better." Assessment strategies included: assessing patients' physical conditions and end-of-life wishes, patients' and family members' understandings of the disease and prognosis, and their expectations and goals. An important first step for improved care is making explicit the provider's communicating and assessing strategies that facilitate end-of-life decision-making. PMID- 11883617 TI - Phases of the qualitative research interview with institutionalized elderly individuals. AB - The in-depth, open-ended formal interview is a mainstay of qualitative nursing research. However, difficulties with this method of data collection are common with institutionalized elderly individuals. During a qualitative study of urinary incontinence among nursing home residents, six distinct phases of the formal interview were identified: introducing, personalizing, reminiscing, contextualizing, closing, and reciprocating. These phases were discovered in the course of analyzing field notes and verbatim transcripts from open-ended formal interviews with 10 respondents. Allowing qualitative research interviews with institutionalized elderly individuals to unfold in this fashion may help researchers studying this group overcome problems with recruitment, retention, and "thin" data. In turn, more institutionalized elderly individuals would reap benefits associated with participation in research interviews and staff would ultimately have the opportunity to understand and appreciate life in an institution from the resident's perspective. PMID- 11883618 TI - The importance of individualized wheelchair seating for frail older adults. PMID- 11883619 TI - Hot food cart. PMID- 11883620 TI - Guidelines for stage-based supports in Alzheimer's care: the FAST-ACT. Functional Assessment Staging Tool-Action Checklist. PMID- 11883621 TI - Expanding your vision of nursing practice. PMID- 11883622 TI - Stressors, social support, coping, and health dysfunction in individuals with Parkinson's disease. AB - The purposes of this descriptive correlational study were to describe the illness related stressors of individuals with Parkinson's disease and to report the extent to which social support and coping responses predict physical and psychosocial health dysfunction. The sample consisted of 70 men and women, age 44 to 80, residing in the community. Participants reported a variety of illness related stressors, most of which occurred on a daily basis and involved some loss in functional abilities. Less perceived availability of social support, more evasive coping, and more confrontive coping predicted greater health dysfunction, suggesting that these factors may be important to consider when assessing individuals with Parkinson's disease and designing interventions for individuals with functional declines. PMID- 11883623 TI - Give yourself credit for "luck". PMID- 11883624 TI - A follow-up study of beliefs held by parents of children with pervasive developmental delay. AB - PROBLEM: Little is known about the effects of parental beliefs on children with disorders such as pervasive developmental delay (PDD). METHODS: A six-question, semistructured, videotaped interview was used to gather preliminary descriptive data from 44 caregivers regarding beliefs about their PDD children (ages 3-15 years). Children and caregivers were selected by purposive sampling from a preadmission waiting list for a child psychiatric inpatient unit. FINDINGS: While responses varied, most caregivers shared concerns about delayed child development, appropriateness of school placement, and future planning. In addition, some indicated they possess inaccurate beliefs regarding the intention of child behavior as well as the PDD diagnosis. All indicated a need for direction in how to effectively help the children. CONCLUSIONS: Parental responses to the interview questions support the author's clinical observations regarding the need for nurse-conducted parent training programs. Parent training should focus on correcting inaccurate parental beliefs and teaching effective ways to promote child development and manage maladaptive behaviors. PMID- 11883625 TI - Descriptions of self: an exploratory study of adolescents with ADHD. AB - PROBLEM: How do adolescents experience attention deficit hyperactivity disorder (ADHD), and how are those experiences shaped and managed? METHODS: A qualitative exploratory design to investigate how adolescents with ADHD (N = 11; 8 males, 3 females) experience, perceive, and manage ADHD. Findings are based on a subset of data from a large grounded theory study of the experiences of living in a family when a child or adolescent has ADHD. The constant comparative method was used to analyze both individual and focus group interview data. FINDINGS: The primary finding, presented as a hypothesis, suggests that an ADHD adolescent's sense of self is distorted and that the development of self has been disrupted due to the neurobiology of ADHD and environmental factors associated with the difficulties of parenting a difficult child, social role expectations, and modeling. CONCLUSIONS: Therapeutic attention to how the self functions is important in stabilizing and ordering an adolescent's sense of self. Intrapsychic psychotherapy may be an important therapeutic approach in helping adolescents with ADHD develop personal responsibility and increased capacity for empathy. PMID- 11883626 TI - Identification and management of schizophrenia in childhood. AB - TOPIC: The identification and management of schizophrenia in childhood. PURPOSE: To provide an overview of what is currently known about childhood schizophrenia. SOURCES: Published literature and personal observations and experiences. CONCLUSIONS: Early identification and treatment of childhood schizophrenia are critical, and more research and education on the part of all mental health professionals are needed in order to identify, provide treatment, and/or make referrals for children with this serious mental disorder. PMID- 11883627 TI - Pediatric psychiatric emergencies. AB - TOPIC: Pediatric psychiatric emergencies, on the increase in emergency departments, challenge mental health professionals and nurses. PURPOSE: To focus on the information needed to evaluate pediatric psychiatric emergencies and the information-gathering process. SOURCES: Personal observations and experiences, published literature. CONCLUSIONS: The efficiency and efficacy with which pediatric psychiatric evaluations are conducted depend on the evaluator's knowledge of the minimum required information for such assessment and the strategies available for overcoming barriers. PMID- 11883628 TI - Lithium in children and adolescents. PMID- 11883629 TI - Excerpts from the report of the surgeon general's conference on children's mental health: a national action agenda. PMID- 11883630 TI - Protecting the health of older individuals. PMID- 11883631 TI - Effects of the quality of dyadic relationships on the psychological well-being of elderly care-recipients. AB - The purpose of this study was to examine the association of the quality of caregiver relationships with the psychological well-being of elderly care recipients. Sociodemographic variables and characteristics of the care-recipient situation (e.g., self-rated physical health, amount of instrumental support needed) were explored as potential predictors of the psychological well-being of elderly individuals. A secondary analysis of data collected during in-home interviews with 37 community-dwelling older adults revealed no significant correlations between the quality of the primary intimate relationship and any dimension of psychological well-being. However, better self-rated health was associated with fewer depressive symptoms, higher morale, greater life satisfaction, and better quality of life. The more instrumental support needed by an individual, the greater their depressive symptoms and the lower their morale. The findings also revealed that the older the individual was, the greater the depressive symptoms were and the lower life satisfaction became. Self-rated physical health predicted each dimension of psychological well-being. The findings suggest that age, the amount of instrumental support needed, and perceptions of physical health are important indicators of the psychological well being of elderly care-recipients. PMID- 11883632 TI - Factors affecting caregivers' ability to make environmental modifications. AB - This study explored factors that family caregivers described as affecting their ability to use environmental modifications. Intensive interviews and participant observation were used to collect detailed data from 24 primary family caregivers. Several factors that affect the caregivers' ability to implement modification strategies were identified in the analysis. These factors included attributes of the elderly individual, attributes of the modification, quality of the caregiver elderly relationship, caregivers' skills, personal resources of the caregiver, and the informal and formal supports available. Of these factors, the most important were the salient skills that caregivers need to implement environmental modifications. These findings point to the importance of caregivers receiving skills training in this important dimension of caregiving. Intervention should be based on a collaborative approach that ensures the caregiver and care receiver's needs and preferences are respected. PMID- 11883633 TI - Translating research into achieving objectives. PMID- 11883634 TI - Purrfect friends. PMID- 11883635 TI - Nutrition and hydration. PMID- 11883636 TI - Use of multidimensional assessment to provide testimony on behalf of residents in a life-care home. AB - City administrators challenged a life-care home's tax-exempt status. A successful, empirically-based case was made based on data collected by gerontological nurses using the Iowa Self-Assessment Inventory (ISAI) and related instruments (Mini-Mental State Examination [MMSE], Instrumental Activities of Daily Living Scale [IADLS], and Physical Self-Maintenance Scale [PSMS]) to describe the life-care population. Testimony by gerontological nurses included comparisons between these life-care residents and statewide data on elderly individuals currently residing in nursing homes and in the community. The data they presented showed not only that this life-care home provided a high quality of life, but also saved society substantial amounts of money in government-funded services that would otherwise be provided to residents of this home. The judge ruled in favor of continuing tax-exempt status for this life-care home because credible evidence clearly demonstrated that taxpayer savings from the services provided to life-care home residents and the life-care commitment dramatically outweighed the taxpayer costs associated with lost tax revenues. PMID- 11883637 TI - Influence of the structure of drug moieties on the in vitro efficacy of HPMA copolymer-geldanamycin derivative conjugates. AB - PURPOSE: To optimize the structure of geldanamycin (GDM) derivative moieties attached to N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers via an enzymatically degradable spacer. METHODS: HPMA copolymers containing different AR GDM (AR = 3-aminopropyl (AP), 6-aminohexyl (AH), and 3-amino-2-hydroxypropyl AP(OH)) were synthesized and characterized. Their cytotoxicity towards the A2780 human ovarian carcinoma cells was evaluated. RESULTS: The cytotoxic efficacy of HPMA copolymer-AR-GDM conjugates depended on the structure of AR-GDM. Particularly, HPMA copolymer-bound AH-GDM, which possessed the longest substituent at the 17-position, demonstrated the highest efficacy among the polymer-bound GDM derivatives; however the activity of free AH-GDM was lower than that of the other free AR-GDMs. The relative increase of the activity of macromolecular AH-GDM when compared to AP-GDM or AP(OH)-GDM correlated with the enhanced recognition of AH-GDM terminated oligopeptide side-chains by the active site of the lysosomal enzyme, cathepsin B. Drug stability and further stabilization upon binding to HPMA copolymer also contributed to the observed phenomena. CONCLUSIONS: AH-GDM was found to be a suitable GDM derivative for the design of a drug delivery system based on HPMA copolymers and enzymatically degradable spacers. PMID- 11883638 TI - HPLC determination of binding of cisplatin to DNA in the presence of biological thiols: implications of dominant platinum-thiol binding to its anticancer action. AB - PURPOSE: The purpose of this work is to evaluate the extent of the binding of cisplatin (cis-diamminedichloroplatinum(II)) to DNA in the presence and absence of biological thiols, glutathione, and cysteine, and to test the hypothesis whether the platinum-thiol complexes can serve as a drug reservoir for subsequent binding to DNA. METHODS: Reactions of cisplatin (50 microM to 1.0 mM) with calf thymus DNA (870 microM to 6.75 mM) in the presence and absence of glutathione and cysteine (0 to 10 mM) were carried out at pH 4.4, 7.0, and 7.3. Following the reactions, the DNA was enzymatically digested with nucleases, separated by RP HPLC, and analyzed to determine the extent of DNA binding. The method was independently verified by proton NMR measurements. RESULTS: At neutral pH, and equimolar concentrations of DNA and thiols, only a very small amount of platinum (<5%) was coordinated to DNA, and most of the platinum was coordinated to the thiols. At pH 4.4, binding to DNA was dominant over the binding to thiols. No conversion of platinum-thiol to platinum-DNA complexes was observed up to 7 days of incubation. CONCLUSION: At physiological pH, the cisplatin was exclusively coordinated to biological thiols and platinum-DNA was a minor adduct. Data presented in this paper does not support the "drug reservoir" hypothesis. PMID- 11883639 TI - Size-dependency of DL-lactide/glycolide copolymer particulates for intra articular delivery system on phagocytosis in rat synovium. AB - PURPOSE: The present study evaluated the size-dependency of DL lactide/glycolide copolymer (PLGA) particulates for an intra articular delivery system on phagocytosis in the rat synovium after administering directly into the joint cavity. We also investigated the biocompatibility of PLGA particulate systems administered directly into the joint cavity of the rat. METHODS: Fluoresceinamine bound PLGA (FA-PLGA) nanospheres and microspheres were prepared by the modified emulsion solven diffusion method. The suspension of these particulate systems was administered into the rat-joint cavity and the biological action of the synovium was evaluated by histological inspection and fluorescence microscopy. RESULTS: A colloidal suspension of the FA-PLGA nanospheres, with a mean diameter of 265 nm, was phagocytosed in the synovium by the macrophages infiltrated through the synovial tissues. The phagocytosed nanospheres were delivered to the deep underlying tissues. An aqueous suspension of the FA-PLGA microspheres, with a mean diameter of 26.5 microm, was not phagocytosed in the macrophages. The macrophages slightly proliferated in the epithelial lining synovial-cells and the microspheres were covered with a granulation of multinucleated giant cells. The molecular weights of the polymer in these particulate systems were slowly reduced in the synovium. Localize inflammatory responses were almost undetected. CONCLUSIONS: PLGA nanospheres should be more suitable for delivery to inflamed synovial tissue than microspheres due to their ability to penetrate the synovium. PLGA particulate systems with biocompatibility in the joint can provide local therapy action in joint disease in a different manner depending on the size of the system. PMID- 11883640 TI - Intracellular processing of poly(ethylene imine)/ribozyme complexes can be observed in living cells by using confocal laser scanning microscopy and inhibitor experiments. AB - PURPOSE: Critical steps in the subcellular processing of poly(ethylene imine)/nucleic acid complexes, especially endosomal/lysosomal escape, were visualized by using living cell confocal laser scanning microscopy (CSLM) to obtain an insight into their mechanism. METHODS: Living cell confocal microscopy was used to examine the intracellular fate of poly(ethylene imine)/ribozyme and poly(L-lysine)/ribozyme complexes over time, in the presence of and without bafilomycin Al, a selective inhibitor of endosomal/lysosomal acidification. The compartment of complex accumulation was identified by confocal microscopy with a fluorescent acidotropic dye. To confirm microscopic data, luciferase reporter gene expression was determined under similar experimental conditions. RESULTS: Poly(ethylene imine)/ribozyme complexes accumulate in acidic vesicles, most probably lysosomes. Release of complexes occurs in a sudden event, very likely due to bursting of these organelles. After release, poly(ethylene imine) and ribozyme spread throughout the cell, during which slight differences in distribution between cytosol and nucleus are visible. No lysosomal escape was observed with poly(L-lysine)/ribozyme complexes or when poly(ethylene imine)/ ribozyme complexes were applied together with bafilomycin A1. Poly(ethylene imine)/plasmid complexes exhibited a high luciferase expression, which was reduced approximately 200-fold when lysosomal acidification was suppressed with bafilomycin A1. CONCLUSIONS: Our data provide, for the first time, direct experimental evidence for the escape of poly(ethylene imine)/nucleic acid complexes from the endosomal/lysosomal compartment. CLSM, in conjunction with living cell microscopy, is a promising tool for studying the subcellular fate of polyplexes in nucleic acid/gene delivery. PMID- 11883641 TI - Comparative inhibitory effects of different compounds on rat oatpl (slc21a1)- and Oatp2 (Slc21a5)-mediated transport. AB - PURPOSE: The purpose of the present study is to examine the selectivity of various inhibitors towards the rat organic anion transporting polypeptides 1 (Oatp1: gene symbol Slc21a1) and 2 (Oatp2: Slc21a5). METHODS: The inhibitory effects of 20 compounds on the Oatpl-mediated transport of estradiol 17beta-D glucuronide and on the Oatp2-mediated transport of digoxin were examined in cDNA transfected LLC-PK1cells. RESULTS: Among the compounds examined in this study, nonsteroidal anti-inflammatory drugs, deoxycorticosterone. and quinidine preferentially inhibited Oatpl. whereas digoxin, quinine, and rifampicin preferentially inhibited Oatp2 at low concentrations. On the other hand, propionic acid, re-ketoglutarate and p-aminohippurate showed no inhibitory effects on either transporter up to a concentration of 1,000 microM. The Ki values of ibuprofen and quinidine were estimated to be 19 and 13 times lower for Oatpl compared with Oatp2, whereas the values for rifampicin, quinine, and digoxin were 13, 20, and 100< times lower for Oatp2 compared with Oatpl. CONCLUSIONS: At low concentrations, some of the tested inhibitors exert selective inhibition of either Oatpl- or Oatp2-mediated substrate transport. These selective inhibitors may be used at appropriate concentrations to estimate the maximum contribution of Oatp1 or Oatp2 to the total substrate uptake into rat hepatocytes. PMID- 11883642 TI - Proton gradient-dependent transport of valproic acid in human placental brush border membrane vesicles. AB - PURPOSE: To investigate the transport mechanism of valproic acid across the human placenta, we used human placental brush-border membrane vesicles and compared them with that of lactic acid. METHODS: Transport of [3H]valproic acid and [14C]lactic acid was measured by using human placental brush-border membrane vesicles. RESULTS: The uptakes of [3H]valproic acid and [14C]lactic acid into brush-border membrane vesicles were greatly stimulated at acidic extravesicular pH. The uptakes of [3H]valproic acid and [14C]lactic acid were inhibited by various fatty acids, p-chloromercuribenzene sulfonate, alpha-cyano-4 hydroxycinnamate, and FCCP. A kinetic analysis showed that it was saturable, with Michaelis constants (Kt) of 1.04 +/- 0.41 mM and 1.71 +/- 0.33 mM for [3H]valproic acid and [14C]lactic acid, respectively. Furthermore, lactic acid competitively inhibited [3H]valproic acid uptake and vice versa. CONCLUSION: These results suggest that the transport of valproic acid across the microvillous membrane of human placenta is mediated by a proton-linked transport system that also transports lactic acid. However, some inhibitors differentially inhibited the uptakes of [3H]valproic acid and [14C]lactic acid, suggesting that other transport systems may also contribute to the elevated fetal blood concentration of valproic acid in gravida. PMID- 11883643 TI - Permeability profiles of M-alkoxysubstituted pyrrolidinoethylesters of phenylcarbamic acid across caco-2 monolayers and human skin. AB - PURPOSE: The purpose of the present research was to study 10 m-alkoxysubstituted pyrrolidinoethylesters of phenylcarbamic acid-potential local anesthetics. The relationships between the structure of the molecule, its physicochemical parameters (log D(oct), log k, R(M), solubility) were correlated to the permeability data obtained from permeation experiments in Caco-2 monolayers and excised human skin in vitro. METHODS: The extent and mechanism(s) of permeability of the series were studied through a Caco-2 monolayer in the apical-to basolateral (a-b) and basolateral-to-apical (b-a) directions. The MTT test was performed to determine cellular damage. In vitro transdermal permeability data were obtained from permeation experiments on excised human skin by using side-by side chambers. Passive diffusion and iontophoretically enhanced permeability were measured. RESULTS: In Caco-2 monolayers, similar results in the shape of the permeability curves were obtained for the two directions. In the b-a direction, the values of P(app) were approximately 2-6 times greater than in the a-b direction. A plot of drug permeability vs. the number of carbons in the alkoxychain plateaued first, after which the permeability decreased by the increasing lipophilicity of the drug. If the log D(oct) of the ester was > or = 3.4 and the MW > 385 Da, no measurable Caco-2 permeability was found. Cell damage was also higher by the more lipophilic compounds. In excised human skin, the relationship between the passive diffusion of the drugs and the number of carbons in the alkoxychain was parabolic (r2 = 0.95). Introducing low-level electrical current (iontophoresis), transdermal permeability of the more hydrophilic phenylcarbamic acid esters increased clearly. CONCLUSIONS: Lipophilicity and solubility of a compound have crucial roles in the permeation process. A very high lipophilicity has, however, a negative influence on the permeability, both intestinally and transdermally. Iontophoresis significantly increases the diffusion of small and less lipophilic compounds. PMID- 11883644 TI - The potential of chitosan in enhancing peptide and protein absorption across the TR146 cell culture model-an in vitro model of the buccal epithelium. AB - PURPOSE: To investigate the potential of chitosan (CS) to enhance buccal peptide and protein absorption, the TR146 cell culture model, a model of the buccal epithelium, was used. METHODS: The sensitivity of TR146 cells to several CS solutions (different salts with different MW) was investigated by using the MTS/ PMS assay. Permeability studies were performed to determine the enhancing effect of CS glutamate (1, 20, 40, 60, and 100 microg/mL) on the permeability of 3H mannitol and fluorescein isothiocyanate labeled dextrans (FD) with various MW (4.4-19.5 kD) across the cell culture model. RESULTS: Sensitivity of TR146 cells to CS solutions depended on the concentration, the pH, and the type of CS salt. CS glutamate solutions (pH 6.0) were found to be the least harmful. CS glutamate was able to increase the permeability of model substances with MW up to 9.5 kD across the cell model. An enhancing effect was found for CS concentrations of 20 microg/mL and higher, correlating with a decrease in TEER values. The 20 microg/mL CS concentration had a negligible effect on the enzyme activity of the cells as determined by the MTS/PMS assay. CONCLUSIONS: CS glutamate is effective in enhancing the transport of macromolecules across the buccal TR146 cell culture model. Therefore. it might be a promising vehicle for peptide and protein buccal administration. PMID- 11883645 TI - Peptide acylation by poly(alpha-hydroxy esters). AB - PURPOSE: Poly(lactic acid) (PLA) and poly(lactic-co-glycolic acid) (PLGA) microspheres were investigated concerning the possible acylation of incorporated peptides. METHODS: Atrial natriuretic peptide (ANP) and salmon calcitonin (sCT) were encapsulated into PLA and PLGA microspheres. Peptide integrity was monitored by HPLC-MS analysis during microsphere degradation for four weeks. sCT fragmentation with endoproteinase Glu-C was used for identifying modified amino acids. Peptide stability in lactic acid solutions was investigated to elucidate possible mechanisms for preventing peptide acylation. RESULTS: Both peptides were acylated by lactic and glycolic acid units inside degrading microspheres in a time-dependent manner. After 21 days, 60% ANP and 7% sCT inside PLA microspheres were acylated. Fragmentation of sCT with endoproteinase Glu-C revealed that besides the N-terminal amine group, lysine, tyrosine or serine are further possible targets to acylation. Stability studies of the peptides in lactic acid solutions suggest that oligomers are the major acylation source and that lower oligomer concentration and higher pH substantially decreased the reaction velocity. CONCLUSIONS: The use of PLA and PLGA for drug delivery needs substantially more circumspection. As, according to FDA standards. the potential hazards of peptide acylation products need to be assessed, our findings may have significant implications for products already on the market. Techniques to minimize the acylation reaction are suggested. PMID- 11883646 TI - Production and characterization of a budesonide nanosuspension for pulmonary administration. AB - PURPOSE: This study describes the production of a budesonide nanosuspension by high-pressure homogenization for pulmonary delivery from 40 mL up to 300 mL. The aim was to obtain a nanosuspension that can be nebulized and is also long-term stable. METHODS: The nanosuspension was produced by high-pressure homogenization. Particle size analysis was performed by laser diffraction and photon correlation spectroscopy. For further particle characterization, zeta potential was determined. To investigate the aerosolization properties, the nanosuspension was nebulized and afterward analyzed on particle size. RESULTS: It was possible to obtain a long-term stable budesonide nanosuspension. Mean particle size of this nanosuspension was about 500-600 nm, analyzed by photon correlation spectroscopy. Analysis by laser diffraction showed that the diameters 95% and 99% were below 3 microm. Budesonide nanosuspension showed a long-term stability; no aggregates and particle growth occurred over the examined period of 1 year. The PCS diameter before and after aerosolization did not change, and the LD diameters increased negligibly, showing the suitability for pulmonary delivery. The scale-up from 40 mL up to 300 mL was performed successfully. CONCLUSIONS: High-pressure homogenization is a production method to obtain nanosuspensions with budesonide for pulmonary applica- PMID- 11883647 TI - Thermophysical properties of pharmaceutically compatible buffers at sub-zero temperatures: implications for freeze-drying. AB - PURPOSE: To evaluate crystallization behavior and collapse temperature (Tg') of buffers in the frozen state, in view of its importance in the development of lyophilized formulations. METHODS: Sodium tartrate, sodium malate, potassium citrate, and sodium citrate buffers were prepared with a pH range within their individual buffering capacities. Crystallization and the Tg' were detected during heating of the frozen solutions using standard DSC and modulated DSC. RESULTS: Citrate and malate did not exhibit crystallization, while succinate and tartrate crystallized during heating of the frozen solutions. The citrate buffer had a higher Tg' than malate and tartrate buffers at the same pH. Tg' vs. pH graphs for citrate and malate buffers studied had a similar shape, with a maximum in Tg' at pH ranging from 3 to 4. The Tg' maximum was explained as a result of a competition between two opposing trends: an increase in the viscosity of the amorphous phase because of an increase in electrostatic interaction, and a decrease in the Tg' because of an increase in a water concentration of the freeze concentrated solution. CONCLUSION: Citrate buffer was identified as the preferred buffer for lyophilized pharmaceuticals because of its higher Tg' and a lower crystallization tendency. PMID- 11883648 TI - Effect of testosterone suppression on the pharmacokinetics of a potent gnRH receptor antagonist. AB - PURPOSE: The expression of cytochrome P450 enzymes (CYPs) in animals and humans is under complex hormonal regulation. Chronic treatment with drugs that alter sex hormone levels such as GnRH receptor agonists or antagonists may affect the expression of hormone-dependent CYPs, and as a result the pharmacokinetics of drugs metabolized by them. METHODS: Enzyme kinetic parameters were obtained by incubating AG-045572 (0.1-30 microM) with human or rat liver microsomes, or expressed CYP3A4 and CYP3A5. The pharmacokinetics of AG-045572 (10 mg/kg i.v. or 20 mg/kg p.o.) were studied in intact male, female, castrated male and male rats pretreated with AG-045572 for 4 days. RESULTS: AG-045572 is metabolized by CYP3A in both rats and humans. The Km values were similar in male and female human, female rat liver microsomes, and expressed CYP3A4 and CYP3A5 (0.39, 0.27, 0.28, 0.25, and 0.26 microM, respectively). The Km in male rat liver microsomes was 1.5 microM, suggesting that in male and female rats AG-045572 is metabolized by different CYP3A isozymes. The oral bioavailability of AG-045572 in intact male rats was 8%, while in female or castrated male rats it was 24%. Pretreatment of intact male rats with AG-045572 i.m. for 4 days resulted in suppression of testosterone to castrate levels, accompanied by an increase in oral bioavailability of AG-045572 to 27%. In the same experiment, the male-specific pulsatile pattern of growth hormone remained unchanged with slightly elevated baseline levels. CONCLUSIONS: The potent GnRH receptor antagonist AG-045572 is metabolized by hormone-dependent CYP3A. As a result, suppression of testosterone by pretreatment with AG-045572 "feminized" its own pharmacokinetics. PMID- 11883650 TI - Determination of chlorophenols by solid-phase microextraction and liquid chromatography with electrochemical detection. AB - A solid-phase microextraction method has been developed for the determination of 19 chlorophenols (CPs) in environmental samples. The analytical procedure involves direct sampling of CPs from water using solid-phase microextraction (SPME) and determination by liquid chromatography with electrochemical detection (LC-ED). Three kinds of fibre [50 microm carbowax-templated resin (CW-TPR), 60 microm polydimethylsiloxane-divinylbenzene (PDMS-DVB) and 85 microm polyacrylate (PA)] were evaluated for the analysis of CPs. Of these fibres, CW-TPR is the most suitable for the determination of CPs in water. Optimal conditions for both desorption and absorption SPME processes, such as composition of the desorption solvent (water-acetonitrile-methanol, 20:30:50) and desorption time (5 min), extraction time (50 min) and temperature (40 degrees C) as well as pH (3.5) and ionic strength (6 g NaCl) were established. The precision of the SPME-LC-ED method gave relative standard deviations (RSDs) of between 4 and 11%. The method was linear over three to four orders of magnitude and the detection limits, from 3 to 8 ng l(-1), were lower than the European Community legislation limits for drinking water. The method was applied to the analysis of CPs in drinking water and wood samples. PMID- 11883649 TI - Ignoring pharmacokinetics may lead to isoboles misinterpretation: illustration with the norfloxacin-theophylline convulsant interaction in rats. AB - PURPOSE: To investigate the norfloxacin-theophylline convulsant interaction in vivo, with an experimental approach distinguishing between pharmacodynamics and pharmacokinetics contributions to the observed effect. METHODS: Male Sprague Dawley rats (n = 38) were infused each compound separately or in different combination ratios. Infusion was maintained until the onset of maximal seizures. Cerebrospinal fluid and plasma samples were collected for high performance liquid chromatography drug determination. The nature and intensity of the pharmacodynamics interaction between drugs was quantified with an isobolographic approach. RESULTS: Isobolograms suggested a relatively marked antagonism between norfloxacin and theophylline at the cerebrospinal fluid (previously shown to be part of the biophase) and dose levels, but not at the plasma (free and total concentrations) levels. These apparent discrepancies could be explained by nonlinear distribution or/and distribution desequilibrium phenomenon. CONCLUSIONS: These findings showed that the quantitative isobolographic approach is appropriate to assess the nature and intensity of the pharmacodynamic interaction between two drugs when data are collected within the biophase, but that data interpretation outside the biophase can be risky due to further pharmacokinetic complexities, in particular slow or/and nonlinear diffusion into the biophase. PMID- 11883651 TI - Comparison of retention of aromatic hydrocarbons with polar groups in binary reversed-phase high-performance liquid chromatography systems. AB - The retention of aromatic hydrocarbons with polar groups has been correlated as log k1 versus log k2 for reversed-phase high-performance liquid chromatography systems with different binary aqueous mobile phases containing methanol, acetonitrile or tetrahydrofuran as modifiers. Distinct changes in separation selectivity have been observed between tetrahydrofuran and acetonitrile or methanol systems. Methanol and acetonitrile systems show lower diversity of separation selectivity. The changes in retention and selectivity of aromatic hydrocarbons with various polar groups between any two chromatographic systems with binary aqueous eluents (tetrahydrofuran vs. acetonitrile, tetrahydrofuran vs. methanol and methanol vs. acetonitrile) have been interpreted in terms of molecular interactions of the solute with especially one component of the stationary phase region, i.e. extracted modifier, and stationary phase ordering. The ordering of the stationary phase region caused by modifier type influences the chromatographic selectivity of solutes with different molecular shape. PMID- 11883652 TI - Fabrication and characterization of a rigid magnetic matrix for protein adsorption. AB - This article describes the fabrication and characterization of a novel magnetic poly(glycidyl methacrylate-triallyl isocyanurate-divinylbenzene) matrix containing magnetite colloids. The results showed that the matrix was superparamagnetic and could be separated magnetically from a suspension in a few seconds. Protein adsorption properties of diethylamine-derivatized matrix were characterized with bovine serum albumin (BSA) as a model protein. The static capacity determined by batch adsorption was 79 mg/ml wet matrix. Kinetic study gave an effective diffusivity of BSA of 5.0 x 10(-13) m2/s in the matrix at an initial BSA concentration in the liquid phase of 1.0 mg/ml. Stability of the matrix was confirmed by recycling of the matrix in protein adsorptions. PMID- 11883653 TI - Synthesis and characterization of endcapped C18 stationary phases using a silica hydride intermediate. AB - The effect of endcapping on an octdecyl bonded phase synthesized by the silanization/hydrosilation method is investigated. The endcapping reagent is a 1:1 molar ratio of trimethylchlorosilane (TMCS) and hexamethyldisilizane (HMDS). Two approaches for endcapping are possible for this synthetic method that produces a silica hydride intermediate: bonding of TMCS-HMDS after silanization (on the hydride intermediate) or after hydrosilation (on the C18 product stationary phase). The use of TMCS-HMDS is designed to eliminate the few remaining silanols on the silica hydride intermediate. The endcapping process is characterized spectroscopically by diffuse reflectance infrared Fourier transform (DRIFT), 29Si cross polarization magic angle spinning nuclear magnetic resonance spectroscopy (CP-MAS-NMR) and 13-C-CP-MAS-NMR. The octadecyl bonded phases are characterized chromatographically by measuring the capacity factors of several hydrophobic and basic test solutes as well as the separation factors among various solute pairs. Finally, long-term stability tests are done on both products at high and low pH. PMID- 11883654 TI - Determination of strong binding chelators and their metal complexes by anion exchange chromatography and inductively coupled plasma mass spectrometry. AB - Based on the negative charge of polycarboxylic chelators, an anion-exchange separation has been developed that is compatible with sensitive metal detection by ICP-MS. A low capacity hydrophilic polymer (AS11) was used as the anion exchanger and ammonium nitrate as the eluent. The new procedure provided high selectivity in the isocratic mode as well as a large separation window and high separation efficiency in the gradient mode. This was demonstrated for different types of chelators and their metal complexes. The aminopolycarboxylates NTA, EDTA, CDTA, DTPA, EDDS and for the EDTA derivatives HEDTA, ED3A and EDTMP, the phosphonic acid analogue of EDTA were tested. Their retention times generally depended on the charge, which was lower in 1:1 metal chelator complexes. Evaluation of the separation mechanism demonstrated that they were all separated predominantly by an anion-exchange mechanism with only a minor contribution from hydrophobic attraction. The method is useful for species identification and for predicting the charge of unknown analogous species from retention times. A gradient separation procedure achieved on-column preconcentration and matrix removal for the interference-free detection of metal chelates down to low nanomolar concentration in samples from various fields of environmental research. PMID- 11883655 TI - Determination of the amount of wash amines and ammonium ion in desulfurization products of process gases and results of related studies. AB - This paper describes a method for the determination of the so-called wash amines and their degradation products, including ammonium ions, in process liquids and wastewater generated during the desulfurization of hydrogen sulfide gas in the process of crude oil refining and also reports the results of related studies. Ion-exchange liquid chromatography employing an inexpensive cation-exchange HPLC column and refractometric detection was used. The results obtained were compared with those obtained by potentiometric titration. Analytical characteristics and a description of the developed procedure are provided. Examples of the results of routine determinations of amines, their degradation products and ammonium ions in process liquids and wastewater are given. PMID- 11883656 TI - Determination of fungicide residues in fruits and vegetables by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. AB - A liquid chromatography (LC) method for the quantitative determination of five fungicide residues (dichloran, flutriafol, o-phenylphenol, prochloraz and tolclofos methyl) in oranges, lemons, bananas, peppers, chards and onions is described. The residues were extracted by matrix solid-phase dispersion (MSPD) using C8. Quantitative analysis was performed by isocratic LC coupled to quadrupole mass spectrometer using atmospheric pressure chemical ionization in the negative ionization mode. The limit of quantification was 0.01 mg kgmicro for flutriafol, o-phenylphenol and dichloran, and 0.1 mg kg(-1) for prochloraz and tolclofos methyl. The MSPD method is also suitable for LC-UV analysis but higher limits of quantification (between 1 and 5 mg kg(-1)) were obtained. Validation of the method was performed between 0.01 and 25 mg kg(-1). Recoveries for fungicides ranged from 52.5 to 91.1% with relative standard deviations between 6.1 and 11.9%. The method was applied to the determination of residues in samples taken from agricultural cooperatives. The fungicides most often detected were o phenylphenol and prochloraz. PMID- 11883657 TI - Rapid separation of microcystins and nodularin using a monolithic silica C18 column. AB - A monolithic C18-bonded silica rod column (Merck Chromolith) was compared to particle-based C18 and amide C16 sorbents in the HPLC separation of eight microcystins and nodularin-R. Two gradient mobile phases of aqueous trifluoroacetic acid modified with acetonitrile or methanol, different flow-rates and different gradient lengths were tested. The performance of the Chromolith column measured as the resolution of some microcystin pairs, the selectivity, efficiency (peak width) and peak asymmetry equalled, or exceeded, the performance of traditional particle-based columns. The Chromolith column allowed a shortening of the total analysis time to 4.3 min with a flow-rate 4 ml min(-1). PMID- 11883658 TI - Simultaneous determination of reboxetine and O-desethylreboxetine enantiomers using enantioselective reversed-phase high-performance liquid chromatography. AB - Current knowledge of stereoselective pharmacokinetics and different potencies of drug enantiomers requires the performance of stereoselective analysis during therapeutic drug monitoring in clinical practice. However, in the case of the new antidepressant drug reboxetine, no effort has been made so far to find a such a suitable system. Therefore, as a step towards developing an enantioselective bioanalytical method for reboxetine and the O-desethylreboxetine metabolite, three stereoselective chromatographic approaches have been investigated. Several chiral columns were tested, among them Chiral-AGP, ChiraGrom 2 and Chiral-CBH, which were able to simultaneously separate the two compounds into enantiomers in total running times of 28, 18 and 12 min, respectively. PMID- 11883659 TI - High-temperature gas chromatography-mass spectrometry with glass capillary columns for the screening of natural products. AB - High-temperature high resolution gas chromatography (HT-HRGC) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS) are powerful but relatively unexplored tools for the analysis of crude extracts and fractions of natural products. To illustrate the scope of the technique the direct characterization of several compounds, present in crude extracts of leaves and stems of Croton hemiargyreus Muell. Arg. var. hemiargyreus was undertaken, without derivatization or clean-up procedures. Both practical aspects and limitations of HT-HRGC and HT-HRGC-MS were evaluated resulting in a simple, straightforward and extremely powerful technique for the analysis of complex mixtures. PMID- 11883660 TI - Analysis of free and bound volatiles by gas chromatography and gas chromatography mass spectrometry in uncased and cased tobaccos. AB - The free and bound volatiles of tobaccos were analyzed by capillary GC and GC-MS. Bound volatiles were isolated by dichloromethane extraction followed by stream distillation continuous extraction (SDE) at pH 2.5 acid hydrolysis. The bound aromatic compounds were hydrolyzed by acid at pH 2.5, and the bound volatiles were liberated and extracted into dichloromethane by SDE simultaneously. In total, 23 volatiles were identified, with neophytadiene, 2-ethyl hexanol, damascenone, benzene ethanol, palmitic acid, stearic acid, linoleic acid, farnesyl acetone, 3-oxo-ionol, and megastigmatrienone being the major components. They consisted mainly of compounds exhibiting aromatic characteristics. The quality and quantity of free and bound volatiles exhibited different distributions in uncased or cased tobaccos. The volatiles existed in higher amounts in bound form than in free form. Compared with uncased tobaccos, free form volatiles showed a decrease after the casing process, while bound volatiles showed an increase. PMID- 11883662 TI - Effect of structure modification of chondroitin sulfate C on its enantioselectivity to basic drugs in capillary electrophoresis. AB - The effect of structure modification of chondroitin sulfate C on its enantioselectivity to several representative basic drugs in capillary electrophoresis was investigated. Chemical desulfation showed no remarkable decrease in selectivity, whereas depolymerization with chondroitinase ABC resulted in complete loss of selectivity. Comparison with chondroitin sulfate A indicated considerable decrease in selectivity with this isomer. The great retention of enantioselectivity in the desulfated derivative suggests that the selectivity comes from the difference of the magnitude of an interaction in the multipoint mechanism between a part of the drug molecule and a functional group in chondroitin sulfate C other than the sulfate group. The sulfate group is not considered to play a major role for chiral separation. The complete loss of selectivity by depolymerization is consistent with a general tendency of lower selectivity in smaller saccharides, and the priority of chondroitin sulfate C to chondroitin sulfate A suggests the importance of the hydroxyl group at C4 in the galactosamine residue. During the course of this work we observed heavy tailing of the peaks of basic drugs in some batches of uncoated fused-silica capillaries under acidic conditions and solved this problem by doubly coating capillaries with Polybrene followed by chondroitin sulfate C. On the other hand, we demonstrated the usefulness of a special technique which uses a short, wider bore PTFE tube-attached capillary for the study of the effect of depolymerization, in order to minimize sample amount. PMID- 11883661 TI - Protein separation and surfactant control of electroosmotic flow in poly(dimethylsiloxane)-coated capillaries and microchips. AB - A thermally pyrolyzed poly(dimethylsiloxane) (PDMS) coating intended to prevent surface adsorption during capillary electrophoretic (CE) [Science 222 (1983) 266] separation of proteins, and to provide a substrate for surfactant adsorption for electroosmotic mobility control was prepared and evaluated. Coating fused-silica capillaries or glass microchip CE devices with a 1% solution of 100 cSt silicone oil in CH2Cl2, followed by forced N2 drying and thermal curing at 400 degrees C for 30 min produced a cross-linked PDMS layer. Addition of 0.01 to 0.02% Brij 35 to a 0.020 M phosphate buffer gave separations of lysozyme, cytochrome c, RNase, and fluorescein-labeled goat anti-human IgG Fab fragment. Respective plates/m typically obtained at 20 kV (740 V cm(-1)) were 2, 1.5, 1.25, and 9.4-10(5). In 50 mM ionic strength phosphate, 0.01% Brij 35 running buffer, the electroosmotic flow observed was about 25% of that in a bare capillary, and showed no pH dependence between pH 6.3-8.2. Addition of sodium dodecylsulfate (SDS) or cetyltrimethylammonium bromide (CTAB) to this running buffer allowed ready control of electroosmotic mobility, mu(eo). Concentrations of SDS between 0.005 to 0.1% resulted in mu(eo) ranging from 3 to 5 x 10(-4) cm2 V(-1) s(-1). Addition of 1 to 2.3 x 10(-4)% (2.7-6.3 microM) CTAB caused flow reversal. CTAB concentrations between 3.5 x 10(-4) and 0.05% (0.0014-1.37 mM) allowed control of mu(eo) between -1 x 10(-4) and -5.0 x 10(-4) cm2 V(-1) s(-1). For both surfactants the added presence of 0.01% Brij 35 provided slowly varying changes in mu(eo) with charged surfactant concentration. PMID- 11883663 TI - Enantioselective separation of racemic secondary amines on a chiral crown ether based liquid chromatography stationary phase. AB - The first general enantioselective separation of racemic secondary amines on a crown ether-based liquid chromatography chiral stationary phase (CSP) is presented. The CSP is based on (+)- or (-)-(18-crown-6)-2,3,11,12-tetracarboxylic acid covalently bonded to silica gel. A mobile phase containing methanol, acetonitrile, triethylamine and acetic acid was employed in these separations of secondary amines with crown ether CSPs. The separation mechanism is believed to be the secondary amine forming a complex which includes crown ether coordination and electrostatic interaction of the positively charged amine with a carboxylate anion of the immobilized crown ether. PMID- 11883664 TI - Detection and quantitation of lactoferrin in bovine whey samples by reversed phase high-performance liquid chromatography on polystyrene-divinylbenzene. AB - An existing RP-HPLC method for the measurement of the major bovine whey proteins in bovine whey samples and powders has been extended to include the analysis of the minor bovine whey protein lactoferrin. Lactoferrin could be detected and quantitated at levels down to 0.2 microg and linear calibration was observed between 0.2 and 30 microg. Reliable quantitation of lactoferrin in whey samples could be achieved provided the bovine serum albumin to lactoferrin ratio did not exceed 10:1. Quantitative data obtained by the RP-HPLC method compared favourably with data obtained by Mono S analytical chromatography. PMID- 11883666 TI - Analysis by gas chromatography-mass spectrometry of the volatile components of Ageratina adenophora Spreng., growing in the Canary Islands. AB - The essential oil of Ageratina adenophora Spreng., growing in the Canary Islands was analyzed by GC-MS. A total of 78 volatile compounds was identified and p cymene (11.6%) was the major component in the oil. The sesquiterpene fraction (44.3%) was higher than the monoterpene one (32.1%). PMID- 11883665 TI - Determination of liposoluble vitamins in cooked meals, milk and milk products by liquid chromatography. AB - A method for the simultaneous determination of liposoluble vitamins in cooked meals was established. Saponification was performed with 50% (w/v) KOH at 80 degrees C, and ascorbic acid was added as antioxidant. The subsequent extraction was carried out with diethyl ether. This was followed by a liquid chromatographic separation on a reversed-phase C18 column with methanol-water (94:6, v/v as the mobile phase. Retinyl acetate was used as the internal standard. The analytical parameters linearity, detection limit (0.19 and 8.33 microg/100 g for retinol and alpha-tocopherol, respectively), precision of the method (RSD=5.24 and 6.99% for retinol and alpha-tocopherol, respectively) and recovery assays (95.6 and 96.5% for retinol and alpha-tocopherol, respectively) show that the method studied is useful for measuring these compounds in foods and cooked meals. PMID- 11883667 TI - Neuropsychological findings in congenital and acquired childhood hydrocephalus. AB - Hydrocephalus is an increase in cerebrospinal fluid volume that can be caused by a variety of etiologies. The most common connatal and acquired causes of hydrocephalus are spina bifida, aqueduct stenosis, and preterm low birthweight infants with ventricular hemorrhage. In general, the literature suggests mild neuropsychological deficits associated with hydrocephalus, which are predominant in visuospatial and motor functions, and other nonlanguage skills. Although the precise nature of the neuropsychological deficits in hydrocephalus are not completely known, several factors such as etiology, raised intracranial pressure, ventricular size, and changes in gray and white matter tissue composition as well as shunt treatment complications have been shown to influence cognition. In fact, the presence of complications and other brain abnormalities in addition to hydrocephalus such as infections, trauma, intraventricular hemorrhage, low birthweight, and asphyxia are important determinants of the ultimate cognitive status, placing the child at a high risk of cognitive impairment. PMID- 11883668 TI - Neuropsychological aspects of pediatric sickle cell disease. AB - Sickle cell disease (SCD), a class of genetic disorders characterized by abnormal, sickled red blood cells, is a chronic illness that results in progressive cerebrovascular disease. Neurocognitive sequelae of clinically apparent cerebrovascular accidents in children with SCD are characterized by pervasive impairments, including decrements in general intellectual functioning, language and verbal abilities, visual-motor and visual-spatial processing, memory, academic achievement, and processing of subtle prosodic information. In contrast, subtle neurocognitive deficits in the areas of attention and concentration, executive function, and visual-motor speed and coordination appear to be associated with silent infarcts that are not necessarily detected on physical examination. Investigation of the disease course and associated neurocognitive sequelae suggest a disease-specific model of neuropsychological impairment. Recommendations are made for clinical and research efforts in the field of pediatric neuropsychology. PMID- 11883670 TI - Inhibition of bacterial foodborne pathogens by the lactoperoxidase system in combination with monolaurin. AB - The lactoperoxidase system (LPS) and monolaurin (ML) are potential natural antimicrobial agents for use in foods. The LPS is considered to have greatest activity against Gram-negative bacteria while ML is usually considered to have greatest activity against Gram-positive bacteria. An LPS-ML combination system (utilizing lactoperoxidase (LPX) in the range 5-200 mg kg(-1) and ML in the range 50-1,000 ppm) inhibited growth of Escherichia coli O157:H7 and Staphylococcus aureus. Growth of S. aureus was inhibited more strongly in broth than in milk, in milk than in ground beef A similar pattern was observed for E. coli O157:H7, though enhanced inhibition by LPS-ML systems over that obtained in comparable LPS only systems was not observed in ground beef The inhibitory action of the LPS in combination with other lipids was also examined, with progressively weaker inhibition observed in combinations including palmitoleic acid, monopalmitolein, lauric acid, caprylic acid, and sodium lauryl sulphate. PMID- 11883669 TI - Approaches to cognitive remediation of neuropsychological deficits in schizophrenia: a review and meta-analysis. AB - A review and critique of the literature pertaining to the use of cognitive remediation techniques in patients with schizophrenia is presented. The review is organized into three sections, according to the neuropsychological deficit targeted for remediation: 1) executive-function, 2) attention, and 3) memory. With regards to executive-function, despite an initial report suggesting that Wisconsin Card Sorting Test performance cannot be remediated, subsequent studies suggest that performance can be improved on a variety of dependent measures including perseverative errors, categories achieved, and conceptual level responses. These observations were confirmed by a meta-analytic investigation that revealed large mean effects sizes (d+ = 0.96) for these studies. Effect sizes were homogenous across discrepant remediation strategies and dependent measures. With regards to attention, serial scanning can be improved with instruction and reinforcement, whereas there is mixed evidence suggesting that practice-based attention drills can improve performance on measures of sustained attention in schizophrenia. With regards to memory, relatively simple semantic and affective elaborate encoding strategies elevates verbal list-learning memory in patients with schizophrenia to levels consistent with controls. A similar encoding procedure, combined with vigilance training, produces substantial improvement in social cue recognition. Avenues for future research are discussed. PMID- 11883671 TI - An emission pattern of a thermophilic bacteria attached to or imbedded in porous supports. AB - There are many problems with thermophilic bacteria contamination of milk in the dairy industry. This is, in part, a result of fouling by milk components on stainless steel surfaces, which provide good harboring facilities for these bacteria to attach, imbed and grow. The interactions between milk fouling and bacteria deposited in or on the fouling deposit therefore become important issues. There have been a number of previous studies on the biofilm development in dairy processing plants. Here, a different approach to investigate the bacteria emission from a porous layer has been taken. In this approach, various process fluids were flushed over the top of a model milk foulant layer that contains high percentages of milk proteins, fat and some bacteria cells, in order to investigate the behavior of the 'resident' microorganisms and how they are 'released' into the flushing liquids. Definitive results were obtained, which have created sufficient interest for a different approach taken later, where fabric layers were used as the support for the bacteria cells to explore the 'generic' behavior of the porous layer-bacteria system. This study has shown that Bacillus stearothermophilus could multiply on or within a porous layer and 'migrate' from the layer into the fluid during processing. This "migration" is somewhat peculiar in terms of its time-responses but these are reproducible in all the tests performed. The phenomena observed may have an impact on future microbial safety practice in food factories. PMID- 11883672 TI - Production of Bacillus cereus emetic toxin (cereulide) in various foods. AB - To determine the role of Bacillus cereus as a potential pathogen in food poisoning, the production of an emetic toxin (cereulide) by B. cereus was quantified in various food sources. The amount of emetic toxin in 13 of 14 food samples implicated in vomiting-type food poisoning cases ranged from 0.01 to 1.28 microg/g. A vomiting-type strain, B. cereus NC7401, was inoculated into various foods and incubated for 24 h at 20, 30, and 35 degrees C. In boiled rice, B. cereus rapidly increased to 10(7)-10(8) cfu/g and produced emetic toxin at both 30 and 35 degrees C. In farinaceous foods, the production of emetic toxin was as high as that in the food samples implicated in food poisoning. Low levels of emetic toxin were detectable in egg and meat and their products and a small quantity of toxin was detectable in liquid foods such as milk and soymilk when not aerated. Bacterial growth and toxin production was inhibited in foods cooked with vinegar, mayonnaise, and catsup, supposedly by the decreased pH of acetic acid. This is the first report that has quantified emetic toxin of B. cereus in various foods. PMID- 11883673 TI - The survival of hepatitis A virus in fresh produce. AB - Fresh produce has been repeatedly implicated as the source of human viral infections, including infection with hepatitis A virus (HAV). The objective of the present study was to evaluate the HAV adsorption capacity of the surface of various fresh vegetables that are generally eaten raw and the persistence of the HAV. To this end, the authors experimentally contaminated samples of lettuce, fennel, and carrot by immersing them in sterile distilled water supplemented with an HAV suspension until reaching a concentration of 5 log tissue culture infectious dose (TCID50)/ml. After contamination, the samples were stored at 4 degrees C and analysed at 0, 2, 4, 7, and 9 days. To detect the HAV, RT-nested PCR was used; positive samples were subjected to the quantitative determination using cell cultures. The three vegetables differed in terms of their adsorption capacity. The highest quantity of virus was consistently detected for lettuce, for which only a slight decrease was observed over time (HAV titre = 4.44 +/- 0.22 log TCID50/ml at day 0 vs. 2.46 +/- 0.17 log TCID50/ml at day 9, before washing). The virus remained vital through the last day of storage. For the other two vegetables, a greater decrease was observed, and complete inactivation had occurred at day 4 for carrot and at day 7 for fennel. For all three vegetables, washing does not guarantee a substantial reduction in the viral contamination. PMID- 11883674 TI - Evaluation of selective enrichment PCR procedures for Yersinia enterocolitica. AB - Four enrichment PCR protocols for detecting unlysed cells of pathogenic Yersinia enterocolitica were studied. First, the probability of detecting Y. enterocolitica cells of known concentrations by a multiplex PCR assay was determined, and it was found to follow a logistic regression model. From this model, the probability of detecting Y enterocolitica at a specific concentration could be estimated; for example, the detection probability of 10(4) CFU/ml was estimated to be 85.4%. The protocols were evaluated on enrichment cultures inoculated with 10(2) CFU/ml Y. enterocolitica and 10(2)-10(6) CFU/ml of a defined background flora. For each protocol, the time for sample withdrawal and the presence of background flora were studied with respect to PCR detection. The optimal point in time of sample withdrawal was found to be different for each protocol employed. Early detection was favoured by concentrating the target cells, and the most rapid PCR detection of Y. enterocolitica was achieved with enrichment in Yersinia-PCR-compatible-enrichment (YPCE) medium for 3 h at 25 degrees C, followed by a centrifugation prior to PCR analysis. For detection of Y. enterocolitica in the presence of high concentrations (10(6) CFU/ml) of background flora, a long incubation time followed by density centrifugation and a dilution step was most successful. The protocol that gave the most reliable PCR detection in the presence of 10(6) CFU/ml background flora included 24 h incubation in Yersinia-selective-enrichment (YSE) broth at 25 degrees C, followed by Percoll density centrifugation, and a 100 times dilution prior to PCR analysis. PMID- 11883675 TI - Fermentation and microflora of plaa-som, a thai fermented fish product prepared with different salt concentrations. AB - Plaa-som is a Thai fermented fish product prepared from snakehead fish, salt, palm syrup and sometimes roasted rice. We studied the effects of different salt concentrations on decrease in pH and on microflora composition during fermentation. Two low-salt batches were prepared, containing 6% and 7% salt (w/w) as well as two high-salt batches, containing 9% and 11% salt. pH decreased rapidly from 6 to 4.5 in low-salt batches, whereas in high-salt batches, a slow or no decrease in pH was found. Lactic acid bacteria (LAB) and yeasts were isolated as the dominant microorganisms during fermentation. LAB counts increased to 10(8)-10(9) cfu g(-1) and yeast counts to 10(7)-5 x 10(7) cfu g(-1) in all batches, except in the 11% salt batch, where counts were 1-2 log lower. Phenotypic tests, ITS-PCR, carbohydrate fermentations and 16S rRNA gene sequencing identified LAB isolates as Pediococcus pentosaceus, Lactobacillus alimentarius/farciminis, Weisella confusa, L. plantarum and Lactococcus garviae. The latter species was only isolated from high-salt batches. Phenotypic characteristics, ITS-PCR and carbohydrate assimilation identified 95% of the yeasts as Zygosaccharomyces rouxii. It is concluded that the fermentation of plaa som is delayed by a salt-level of 9% due to an inhibition of LAB growth. The growth of Z. rouxii has no influence on the fermentation rate, but may contribute positively to the flavour development of the product. PMID- 11883676 TI - Combined antimicrobial effect of nisin and a listeriophage against Listeria monocytogenes in broth but not in buffer or on raw beef. AB - The effect of nisin and listeriophage LH7, alone and in combination, on the growth and survival of two strains of Listeria monocytogenes in broth and two model food systems, with appropriate controls, was determined. Growth curves for both bacterial strains in tryptic soy broth incubated at 7 or 30 degrees C, and with the addition of nisin and/or listeriophage at lag, mid-exponential or early stationary phase, were obtained by measuring absorbance at 550 nm. Numbers of mixed populations of both L. monocytogenes strains in phosphate buffered saline (pH 5.5) and on vacuum-packaged fresh beef, both stored for 4 weeks at 4 degrees C, and with the addition of nisin and/or listeriophage, were determined. This was achieved by plating appropriately diluted samples on both Tryptic Soy Agar and Modified Oxford Agar to determine both L. monocytogenes numbers and the presence of sub-lethal injury. In broth nisin alone, reduced levels or prevented growth of the two strains under the conditions studied, but regrowth to levels equivalent to those of untreated cells, occurred. Listeriophage LH7 alone, on the other hand, had no effect in broth under the conditions studied. Notably, however, a mixture of nisin and listeriophage displayed a combined effect in broth and reduced levels of cells substantially without regrowth under the conditions studied. In both model food systems only nisin appeared to be active, in a manner consistent with existing literature, and no combined action was apparent. The use of nisin and listeriophage has potential to control L. monocytogenes in foods but a further understanding of the interactions in this complex system needs to be achieved before it could be applied practically. PMID- 11883677 TI - Evaluation of antilisterial action of cilantro oil on vacuum packed ham. AB - Cilantro oil is an essential oil preparation extracted from the plant Coriandrum sativium. A series of experiments were conducted to evaluate the ability of cilantro oil to control the growth of Listeria monocytogenes on vacuum-packed ham. The in vitro minimal inhibitory concentration for five strains of L. monocytogenes was found to vary from 0.074% to 0.018% depending on strain. Cilantro oil treatments were then tested on ham disks inoculated with a cocktail of the five L. monocytogenes strains. The treatments studied were 0.1%, 0.5%, and 6% cilantro oil diluted in sterile canola oil or incorporated into a gelatin gel in which lecithin was used to enhance incorporation of the cilantro oil. Gelatin gel treatments were also conducted with 1.4% monolaurin with or without 6% cilantro oil to determine if an interaction between the antimicrobials could increase inhibition of L. monocytogenes. Treated ham was then vacuum-packed and stored at 10 degrees C for up to 4 weeks. The only cilantro oil treatment which inhibited growth of L. monocytogenes on the ham samples was 6% cilantro oil gel. Samples receiving this treatment had populations of L. monocytogenes 1.3 log CFU/ml lower than controls at week 1 of storage, though there was no difference between treatments from week 2 onward. It appears that immobilization of the antimicrobial in a gel enhanced the effect of treatments. Cilantro oil does not appear to be a suitable agent for the control of L. monocytogenes on ham. The possible reasons for reduced effectiveness of cilantro oil against L. monocytogenes on ham are discussed. PMID- 11883678 TI - Whaling: when is enough, enough? PMID- 11883679 TI - Differences in the acute toxicities of tributyltin between the Caprellidea and the Gammaridea (Crustacea: Amphipoda). AB - Tests for the acute toxicity of tributyltin (TBT) were conducted on amphipod crustaceans collected from Otsuchi Bay, Japan. Five species of caprellids and three species of gammarids, which belong to a closely related ecological niche, were used for the exposure experiments at seven test concentrations (0, 0.001, 0.01, 0.1, 1, 10 and 100 microg TBTCl/l) for 48 h at 20 degrees C. The 48-h LC50 values of the caprellids were 1.2-6.6 microg TBTCl/l, and these were significantly lower than those of the gammarids (17.8-23.1 microg TBTCl/l). This suggests that caprellids are more sensitive to TBT than gammarids. Furthermore, the proportions of TBT and its derivatives, dibutyltin (DBT) and monobutyltin (MBT), were measured in the amphipods collected from Otsuchi Bay. In the caprellids, TBT was the predominant compound, accounting for 72% of the total butyltin which reflected the butyltin ratio in seawater, while in the gammarids, TBT's breakdown products (DBT and MBT) predominated, accounting for 75% of the total butyltin. This difference suggests that caprellids may have lower metabolic capacity to degrade TBT than gammarids. Therefore, the difference in sensitivity to TBT among the amphipods is thought to be related to the species-specific capacity to metabolize TBT. PMID- 11883680 TI - Experimental transplanting of Posidonia australis seagrass in Port Hacking, Australia, to assess the feasibility of restoration. AB - Over the last 50 years, about one-third of the original area of the seagrass Posidonia australis has been lost from Port Hacking (Australia) due to anthropogenic impacts. To assess the feasibility of restoring these seagrass meadows, healthy Posidonia rhizomes were transplanted to four impact sites and one control site. Survival rates of transplanted shoots were monitored in situ bi monthly for 16 months and, at the end of the experiment, rhizome growth, shoot growth, shoot production and growth architecture were assessed by harvesting tagged rhizomes. A total of 575 shoots were transplanted and after 16 months 650 shoots were present. Four of the five sites exhibited high survival rates in the short term (less than six months) but only two impact sites, Burraneer Bay (BB) and Red Jacks Point (RJP), and the control site (CS) survived to the end of the experiment. Total number of shoots increased by 61% at CS, tripled at BB, but decreased by 22% at RJP. Rhizome growth varied significantly between site, from 22.3 +/- 1.4 cm yr(-1) at BB to 9.1 +/- 1.0 cm yr(-1) at RJP. Shoot growth did not vary significantly between sites and was approximately 2-3 cm yr(-1). At BB and CS there was substantial colonisation of the surrounding substrate, with new rhizomes, orthotropic shoots and transitional shoots produced. Survival of transplants appeared to depend on whether the factors that had caused the original loss of Posidonia were still operating in the study area. PMID- 11883681 TI - Biological pollution in the Mediterranean Sea: invasive versus introduced macrophytes. AB - The authors have listed 85 species of macrophytes that have probably been introduced to the Mediterranean. Among them, nine species can be considered as invasive, i.e., playing a conspicuous role in the recipient ecosystems, taking the place of keystone species and/or being economically harmful: Acrothamnion preissii, Asparagopsis armata, Lophocladia lallemandii, Womersleyella setacea (Rhodophyta), Sargassum muticum, Stypopodium schimperi (Fucophyceae), Caulerpa racemosa, Caulerpa taxifolia and Halophila stipulacea (Plantae). These data fit well the Williamson and Fitter's "tens rule", which states that, on average, 1 out of 10 introduced species becomes invasive. Though some features (e.g. life traits, geographical origin) can increase the likelihood of a successful invasion, the success of invaders is far from being predictable. Since the beginning of the 20th century, the number of introduced species to the Mediterranean has nearly doubled every 20 years. Should these kinetics continue, and according to the tens rule, it can be expected that 5-10 newly introduced macrophytes shall become invasive in the next 20 years. PMID- 11883682 TI - Influence of the Aznalcollar mining spill on the vertical distribution of heavy metals in sediments from the Guadalquivir estuary (SW Spain). AB - The Natural Park of Donana and the Guadalquivir estuary were impacted by the release of 6 million cubic meters of acid waste after the mine-tailing spill in Aznalcollar (Andalusia, SW, Spain). Here is presented the monitoring of the accidental spill on vertical distribution of heavy metals in the estuarine sediments. The total concentration of six metals (Fe, Mn, Zn, Cd, Pb, Cu), their chemical speciation and the organic carbon concentration were analyzed in sediment vertical profiles. The results obtained determine background levels similar to previously reported in the area. The analysis catalogues the impact of the accident on the estuary as acute and mainly associated with high concentrations of Zn and Cd. The recent enrichment in Zn and Cd and their geochemical association with the more mobile fractions of the sediment determine an environmental risk associated with the acute impact and detected in some of the areas of the estuary. PMID- 11883683 TI - Petroleum and PAH contamination of the Black Sea. AB - Concern has been expressed regarding the extent of contamination of the Black Sea. Analyses of coastal sediments taken from throughout the region indicate, however, that levels of petroleum hydrocarbons (2-300 microg g(-1) dry wt total hydrocarbons) are generally comparable to those encountered in the Mediterranean and are lower than concentrations reported for highly contaminated areas such as the Gulf, Hong Kong, Taiwan and New York Bight. Highest concentrations of total hydrocarbons (>100 microg g(-1) dry wt) were associated with discharges from Odessa, Sochi and the River Danube. Chronic/degraded petroleum was the major contributor at these sites. Samples from the Ukrainian coastline were comparatively clean (<10 microg g(-1) dry wt total hydrocarbons). Major contributions of fresh oil (as indicated by sigma n-C14-34) occur through the River Danube. Concerning total PAH, concentrations (7-638 ng g(-1) dry wt) compare to relatively unpolluted locations in the Mediterranean and are much lower than levels reported for polluted UK estuaries (e.g. Mersey, Tyne, Thames). Both pyrolytic and petrogenic PAH are present in most samples, although petroleum derived PAH are dominant at Sochi and pyrolytic sources are prevalent in the Bosphorus region. The absence of a correlation between total hydrocarbons and PAH (R2 = 0.04) indicates different primary sources for the two. PMID- 11883684 TI - Concordance degrees in macrozoobenthic monitoring programmes using different sampling methods and taxonomic resolution levels. AB - In summer 1997, an intensive survey on the hard bottoms of the 'Abra de Bilbao' (N. Iberian Peninsula) was carried out in the context of the macrozoobenthic monitoring programmes developed to assess the biological recovery of the area. Three types of measurements (abundance, biomass and cover) were used to describe and compare the structure and composition of the communities at three littoral zones: subtidal, lower intertidal and upper intertidal. In addition, several taxonomic aggregation levels of data were successfully applied. The main objective of this paper is to explore the relative effect caused on the results of such programmes by earlier decisions concerning the type of measurement and the taxonomic resolution level to be applied. A 'second stage' multivariate procedure of analysis has been perforrmed based on the previously obtained sampling site ordinations. The measurement type chosen has been found to have a greater effect on the results than the taxonomic resolution used. Moreover, it is suggested that analyses based on abundance data usually lose more information when taxonomic resolution decreases than those based on biomass or cover estimates. The highest concordance among the different analyses has been found in the subtidal zone, which is considered the most appropriate habitat for the development of benthic monitoring programmes. PMID- 11883685 TI - Integrating toxicology and ecology: putting the "eco" into ecotoxicology. AB - Environmental toxicology has been and continues to be an important discipline (e.g., single-species testing for screening purposes). However, ecological toxicology (ecotoxicology--more realism in tests, test species and exposures) is required for predicting real world effects and for site-specific assessments. Ecotoxicology and ecology have shown similar developmental patterns over time; closer cooperation between ecologists and toxicologists would benefit both disciplines. Ecology can be incorporated into toxicology either extrinsically (separately, e.g., providing information on pre-selected test species) or intrinsically (e.g., as part of test species selection)--the latter is preferable. General guidelines for acute and chronic testing and criteria for species selection differ for ecotoxicology and environmental toxicology, and are outlined. An overall framework is proposed based on ecological risk assessment (ERA), for combining ecology and toxicology (environmental and ecological) for decision-making. Increased emphasis on ecotoxicology represents a shift from reductionist to holistic approaches. PMID- 11883686 TI - Hydrocarbon contamination in Cartagena Bay, Colombia. AB - This study deals with the levels of aromatic and aliphatic hydrocarbon quantification in sediments and organisms in Cartagena Bay (Colombia), 1996-1997. Sediments (14 stations) and bivalves (2 stations) were monitored at different times of the year. Areas with high values were in the north with concentrations above 100 microg/g with a maximum of 1415 microg/g. Areas with low values were located toward the south, near the outlet of the Canal del Dique and Baru Island, with values below 10 microg/g. In other areas concentrations were between 50 and 100 microg/g. A decrease in sediment concentrations of hydrocarbons has occurred since 1983, but levels in some sectors are still similar to those in polluted areas. Organisms have relatively low values (8-30 microg/g for bivalves, and 10 40 microg/g for fish). PMID- 11883687 TI - Subcellular distribution of heavy metals in livers and kidneys of Stenella coeruleoalba and Tursiops truncatus from the Mediterranean Sea. PMID- 11883688 TI - Statistical power of non-parametric tests: a quick guide for designing sampling strategies. AB - The importance of considering statistical power in marine pollution studies is unequivocal. However, the vast majority of ecological literature on power analysis focuses on parametric rather than non-parametric tests. This note describes a Monte Carlo simulation method for estimating the power of non parametric tests. The method is illustrated using ordinal data. PMID- 11883689 TI - Priming, boosting, measuring. PMID- 11883690 TI - Human papillomavirus vaccines. AB - Virus-like particle (VLP) subunit vaccines composed of the major capsid protein L1 of the genital human papillomaviruses (HPVs) are under test in phase I and II clinical trials. The vaccines are immunogenic and safe but no data on efficacy are yet available. VLPs induce strong cell-mediated as well as humoral immune responses, and chimeric VLPs, including an HPV early protein, may have therapeutic potential. Polynucleotide and recombinant viral vaccines encoding nonstructural viral proteins show therapeutic and prophylactic efficacy in animal models and are candidate immunotherapies for established low-grade benign genital infections. Recombinant virus, peptide, protein, polynucleotide and dendritic cell vaccines designed to elicit cytotoxic T-lymphocytes specific for the HPV oncoproteins E6 and E7 show immunogenicity and efficacy in transplantable tumor models in rodents. Immunogenicity, but no efficacy, has been demonstrated in small clinical trials with some of these approaches. PMID- 11883691 TI - Prime-boost immunization strategies for infectious diseases. AB - New vaccination strategies that induce the cellular arm of the immune response are needed for the development of effective prophylactic and therapeutic vaccines against a number of intracellular pathogens. DNA vaccines, recombinant viral vectors and recombinant proteins are all effective antigen delivery systems for inducing cellular immunity; however, when used alone, the levels of specific responses they induce are low. Prime-boost immunization strategies involve using two different vaccines, each encoding the same antigen, some weeks apart. Such strategies have been shown to enhance cellular immunity in several different animal and disease models. PMID- 11883692 TI - Alphavirus-based vaccines. AB - Alphavirus vectors can be appliedfor vaccine production as naked RNA molecules, DNA plasmids or recombinant replication-deficient viral particles. The common feature for all these vectors is the alphavirus replicon, which is responsible for the strong RNA amplification in host cells enabling extreme transgene expression levels. Additionally, live replication-proficient virus has elicited reasonable immune responses. For many viral genes, humoral and cellular cytotoxic T-lymphocyte responses have been obtained. In other cases, monoclonal antibodies have been generated against the encoded antigen. Moreover, protection against challenges with lethal doses of virus has been achieved. Alphavirus vaccine applications have been further extended to models for prophylactic tumor therapy. PMID- 11883693 TI - Recombinant yeast as a vaccine vector for the induction of cytotoxic T-lymphocyte responses. AB - There is currently a need to develop safe and effective vaccines for the induction of cellular, in particular cytotoxic T-lymphocyte-mediated, protective immune responses. This review focuses on the challenges presented by such an endeavor, current strategies for the induction of such responses, and the relatively new idea that recombinant yeast may represent an ideal vaccine vector for the induction of cellular immune responses. PMID- 11883694 TI - DNA gene fusion vaccines against cancer. AB - The ability of DNA vaccination to induce effective immune responses has been shown in a range of preclinical disease models. However, the potency of DNA vaccines must be further improved for their use in patients. DNA fusion vaccine strategies, whereby target antigens are genetically linked to immuno-enhancing molecules, are currently being explored. The ease of DNA manipulation has allowed incorporation of a wide variety of molecules able to promote antigen uptake, processing and presentation by professional antigen-presenting cells, to provide critical CD4 T-cell help and to activate more effective immune effector pathways. These strategies are particularly important for cancer vaccines to increase their immunogenicity and to overcome tolerance. PMID- 11883695 TI - Disabled infectious single cycle-herpes simplex virus (DISC-HSV) as a vector for immunogene therapy of cancer. AB - Disabled infectious single cycle-herpes simplex viruses (DISC-HSV) have been shown to be safe for use in humans and may be considered efficacious as vectors for immunogene therapy in cancer. Preclinical studies show that DISC-HSV is an efficient delivery system for cytokine genes and antigens. DISC-HSV infects a high proportion of cells, resulting in rapid gene expression for at least 72 h. The DISC-HSV-mGM-CSF vector, when inoculated into tumors, induces tumor regression in a high percentage of animals, concomitant with establishing a cytotoxic T-cell response, which is MHC class I restricted and directed against peptides of known tumor antigens. The inherent properties of DISC-HSV makes it a suitable vector for consideration in human immunogene therapy trials. PMID- 11883696 TI - DNA-based approaches to the treatment of allergies. AB - Although excellent pharmacological treatments for allergies exist, they do not change the underlying pathogenesis of allergic diseases and do not cure the disease. Only allergen-specific immunotherapy, the injection of small but increasing amounts of allergen, has been shown to change a pre-existing allergic Th2 immune response to a non-allergic Th1 response. However, since injection of allergen is associated with the risk of allergic and sometimes even life threatening anaphylactic reactions, immunotherapy is no longer used as extensively as in the past. In the search for a novel immunotherapy having a low risk-to-benefit ratio, immunostimulatory CpG motif DNA sequences have recently been shown to provide an excellent tool for designing safer and more efficient forms of allergen immunotherapy. These DNA-based immunotherapeutics include allergen gene vaccines, immunization with allergen-DNA conjugates and immunomodulation with immunostimulatory oligodeoxynucleotides. All three DNA based immunotherapeutics have been shown to be very effective in animal models of allergic diseases and, at present, allergen-DNA conjugates are being tested for their safety and efficacy in allergic patients. This review describes the preclinical findings and the data of the first clinical trials in allergic patients of DNA-based immunotherapeutics for allergic disorders. PMID- 11883697 TI - Technology evaluation: BL22, NCI. AB - BL22 (RFB4(dsFv)-PE38) is a recombinant Pseudomonas exotoxin-based immunotoxin under development by the National Cancer Institute for the treatment of B-cell malignancies. It is composed of the disulfide-stabilized Fv portion of the anti CD22 antibody RFB4 genetically fused to a truncated form of Pseudomonas exotoxin A. It has entered phase I trials for the treatment of B-cell lymphoma. PMID- 11883698 TI - Technology evaluation: APC-8015, Dendreon. AB - Dendreon (formerly Activated Cell Therapy), in association with the Mayo Clinic, is developing the dendritic cell therapy APC-8015 (Provenge) for the potential treatment of hormone-refractory prostate cancer [284376]. Phase III trials were initiated in January 2000 [353557], and in July 2001 Dendreon anticipated that preliminary results would be available by the end of the year [417283], [427591]. As of September 2001, Dendreon was planning tofile a BLA in 2002 [421356]. Provenge involves the use of a proprietary recombinant antigen derived from prostatic acid phosphatase, found in approximately 95% of prostate cancers. The target antigen is combined with the patient's own dendritic cells and reinfused into the patient to stimulate an immune response [406383]. In November 1999, Dendreon received US-05976546, which covers the composition of the prostate tumor antigen engineered by Dendreon to help stimulate the immune system [347885]. In August 2000, Dendreon received US-06080409, entitled 'Immunostimulatory composition', which relates to the method by which Dendreon's vaccines stimulate the T-cell arm of the immune system tofight cancer [379085]. In April 2001, Dendreon was awarded US-06210662 covering the therapeutic composition of APC-8015 [406383]. PMID- 11883699 TI - Technology evaluation: Allovectin-7, Vical. AB - Allovectin-7 is a gene transfer product consisting of the human leukocyte antigen HLA-B7 gene co-expressed with the beta2-microglobulin gene. Allovectin-7 is being developed as an immunotherapy approach for a variety of malignancies, with special focus on melanoma, and head and neck cancer. Efficacy results in the phase II setting appear to be promising; an 11% systemic response rate among the intent-to-treat population and 15% response rate in the evaluable population was observed in patients with refractory metastatic melanoma with disease limited to skin, lymph nodes and lung. Stable disease was seen in 19.2% of the intent-to treat population and 25.9% of the evaluable population in the same group of patients. Treatment has been extremely well tolerated with the most common side effects including mild-to-moderate injection site reactions and flu-like symptoms, all of which resolved rapidly and decreased in incidence after the first injection. Current evaluation of the drug includes higher Allovectin-7 doses and injection of multiple tumors in metastatic melanoma patients. A phase III trial comparing Allovectin-7 plus dacarbazine versus dacarbazine in untreated patients with metastatic melanoma has been completed, and preliminary results are soon to be reported. Phase I/II data also indicate promising activity of Alovectin-7 in patients with advanced refractory head and neck squamous cell carcinoma, with 10% of 60 patients achieving partial response and 23% stable disease after one cycle of treatment. Trials of Allovectin-7 as an adjuvant treatment in earlier stages of disease evolution are planned. PMID- 11883701 TI - Interleukin-18 reduces expression of cardiac tumor necrosis factor-alpha and atrial natriuretic peptide in a murine model of viral myocarditis. AB - Heart failure is generally believed to begin with myocyte damage caused by a variety of insults, including ischemia, toxin or myocardial infection. The proinflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), has been hypothesized to play a pathogenetic role in the transition from compensated to decompensated heart failure. Interleukin-18 (IL-18), a recently cloned cytokine synthesized by Kupffer cells, activates macrophages. We examined the therapeutic effect of IL-18 on the modulation of TNF-alpha gene expression in failing heart in a murine model of heart failure caused by viral myocarditis. The heart weight (HW)/ body weight (BW) ratio in IL-18 treated mice 7 days after viral inoculation was significantly lower (P<0.01) than in the untreated controls. Myocardial necrosis and inflammatory cell infiltration were significantly lower in IL-18 treated mice than untreated mice 5 and 7 days after inoculation. The expression of TNF-alpha mRNA in the myocardium was significantly lower on days 5 and 7 in IL 18 treated mice than in infected untreated mice. We conclude that concurrent systemic administration of IL-18 is beneficial in mice with myocarditis, and may be mediated through reduced expression of TNF-alpha in the heart. PMID- 11883702 TI - 4,4'-diisothiocyano-2 ,2'-stilbenedisulfonate protects cultured cerebellar granule neurons from death. AB - We examined the effects of 4,4'-diisothiocyano-2,2'-stilbenedisulfonate (DIDS), an inhibitor of the chloride-bicarbonate exchangers and chloride channels, on death in cultured cerebellar granule neurons. Various stimuli, such as reduction of extracellular K+ concentration, removal of growth factors, and staurosporine treatment, induced cell death. This death was blocked by DIDS in a dose dependent manner. In the presence of DIDS, the cells exposed to such stimuli did not show DNA fragmentation, but retained the ability to exclude trypan blue and to metabolize MTT to formazan. On the other hand, pretreatment of the cells with DIDS did not show any protective effects. The neuroprotective effect of DIDS was not influenced by extracellular Na+, Cl-, HCO3- or Ca2+ concentrations, although reduction of extracellular Cl- or Ca2+ concentrations per se induced neuronal death. Other chloride-bicarbonate exchange blockers like 4-acetamido-4' isothiocyanatostilmene-2,2'-disulfonic acid (SITS) or 4,4'-dinitrostilbene-2,2' disulfonic acid (DNDS) showed no significant effects on neuronal survival under these death-inducing stimuli. Dimethylamiloride, an inhibitor of the Na+/H+ exchanger, did not influence neuronal death induced by these stimuli. Cells undergoing death showed gradual intracellular acidification, and DIDS did not inhibit this response, although DIDS (2 mM) per se induced transitory acidification followed by recovery within 10 min. DIDS did not influence intracellular Ca2+ or Cl-levels during the lethal process. DIDS suppressed the cleavage of caspase-3 in the cells exposed to the death-inducing stimuli. These findings suggest that the neuroprotective effect of DIDS is mediated by a novel mechanism other than by nonselective inhibition of transporters or channels, and that DIDS blocks the death program upstream of caspases and downstream of all of the activation processes triggered by various stimuli. PMID- 11883703 TI - CCK processing by pituitary GH3 cells, human teratocarcinoma cells NT2 and hNT differentiated human neuronal cells evidence for a differentiation-induced change in enzyme expression and pro CCK processing. AB - Human teratocarcinoma Ntera2/c 1.D1 (NT2) cells express very low levels of the prohormone convertase enzyme PC1, moderate levels of PC2 and significant levels of PC5. When infected with an adenovirus which expresses rat CCK mRNA, several glycine-extended forms were secreted that co-eluted with CCK 33, 22 and 12. Amidated CCK is not produced because these cells appear to lack the amidating enzyme. Pituitary GH3 cells express high levels of PC2 and PC5. CCK adenovirus infected GH3 cells secrete amidated versions of the same peptides as NT2 cells. Differentiation of NT2 cells into hNT cells with retinoic acid and mitotic inhibitors increased expression of PC5 and decreased expression of PCI and PC2. CCK adenovirus-infected differentiated hNT cells also secrete glycine extended CCK products and the major molecular form produced co-eluted with CCK 8 Gly. These experiments demonstrate that the state of differentiation of this neuronal cell line influences its expression of PC 1,2, and 5 and its cleavage of pro CCK and suggests that these cells may make an interesting model to study how differentiation alters prohormone processing. These results also support the hypothesis that PC5 in differentiated neuronal cells is capable of processing pro CCK to glycine-extended CCK 8. PMID- 11883704 TI - Involvement of caspase-3 activation in squamocin-induced apoptosis in leukemia cell line HL-60. AB - Annonaceous acetogenins have potent antitumor effect in vitro and in vivo. Squamocin is one of the annonaceous acetogenins and has been reported to have antiproliferative effect on cancer cells. Our results from this study showed that squamocin inhibited proliferation of HL-60 cells with IC50 value of 0.17 microg/ml and induced apoptosis of HL-60 cells. Investigation of the mechanism of squamocin-induced apoptosis revealed that treatment of HL-60 cells with squamocin resulted in extensive nuclear condensation. DNA fragmentation, cleavage of the death substrate poly (ADP-ribose) polymerase (PARP) and induction of caspase-3 activity. Pretreatment of HL-60 cells with caspase-3 specific inhibitor DEVD-CHO prevented squamocin-induced DNA fragmentation, PARP cleavage and cell death. The expression levels of protein bcl-2, bax have no change in response to squamocin treatment in HL-60 cells, whereas stress-activated protein kinase (SAPK/JNK) was activated after treatment with squamocin in HL-60 cells. These results suggest that apoptosis of HL-60 cells induced by squamocin requires caspase-3 activation and is related to SAPK activation. PMID- 11883705 TI - Hyperpnea-induced production of TBHP-initiated chemiluminescence in guinea pigs. AB - Antioxidants attenuate hyperpnea-induced airway constriction. It was hypothesized that this type of airway constriction is closely related to reactive oxygen species (ROS). However, there is no direct evidence of an increase in ROS during or right after the course of hyperpnea. To detect ROS production induced by hyperpnea, forty one guinea pigs were divided into four groups: control; control with 95% O2-5% CO2; hyperpnea with 95% air-5% CO2; and hyperpnea with 95% O2-5% CO2. Three minutes following hyperpnea or at the equivalent time, we obtained bronchoalveolar lavage (BAL) and measured its chemiluminescence (CL) counts. In addition, hyperpnea with 95% O2-5% CO2 gas mixture was carried out and BAL was collected 3 minutes after the hyperpnea in an additional forty animals. We measured CL counts in BAL samples before and after the treatments of the following ROS scavenger(s) or saline in vitro: control (saline); superoxide dismutase (SOD); catalase; dimethylthiourea (DMTU); and SOD+catalase+DMTU. Hyperpnea with 95% O2-5% CO2, but not with 95% air-5% CO2, gas mixture induced significant increase in t-butyl hydroperoxide-initiated CL counts, which were inhibited by DMTU, catalase, or SOD in vitro. Our data suggest that hyperpnea with a 95% O2-5% CO2, but not with 95% air-5% CO2, gas mixture induced an increase in ROS production. PMID- 11883706 TI - Leaf extracts of Carlowrightia cordifolia induce macrophage nitric oxide production. AB - Carlowrightia cordifolia (Acanthaceae) is a medicinal plant used in northeastern Mexico as a traditional remedy against inflammation. As tissue release of nitric oxide (NO) has been correlated with both inflammatory and anti-inflammatory processes, the aim of this study was to determine the effect of C. cordifolia leaf extracts on macrophage NO production. Lipopolysaccharide (LPS)-stimulated and non-LPS-stimulated mouse peritoneal macrophages were incubated with aqueous, ethanol, methanol and hexane extracts of C. cordifolia leaves. All extracts inhibited NO release from LPS-stimulated macrophages, with methanol and hexane extracts showing the greatest inhibition. On the other hand, macrophage cultures treated with extracts without LPS-stimulation produced high releases of NO. These unexpected results suggest two different ways by which leaf extracts may act, depending on cell status. On the other hand, data on NO activity in relation to inflammatory/anti-inflammatory auto-regulatory feedback and high concentrations of NO release by non-stimulated macrophages agreed with the hypothesis that NO may have an inhibitory effect in vascular inflammation. PMID- 11883707 TI - Pharmacological characteristics of the endothelial target for acetylcholine induced vascular relaxation. AB - The pharmacological characteristics of the endothelial target for acetylcholine induced vascular relaxation were investigated in this experiment. The isolated preparations of arteries were suspended for the measurement of isometric force in modified Krebs-Ringer bicarbonate solution (37 degrees C aerated with 95% O2 and 5% CO2). Similar to acetylcholine, carbachol rather than thiocholine, butylcholine and choline could induce endothelium-dependent relaxation. Among cholinergic receptor agonists, arecoline and oxotremorine rather than nicotine could mimic the effects of acetylcholine. But muscarinic agonist pilocarpine had no effect. This phenomenon was observed in rat, cat and rabbit aorta, as well as cat mesenteric. femoral and renal arteries. The new compound tricyclopinate and phenyl cyclopentyl hydroxyl-ethoxy quinuclidines, the competitive antagonists against muscarinic receptors, displayed noncompetitive antagonism against the endothelial target for acetylcholine. Among the six isomers of the novel compound 2-(2'-cyclopentyl-2'-phenyl-2'-hydroxyl-ethoxy) tropane, the isomers with IS 2alpha-2'R and 1S-2alpha-2'S configuration caused the dose-response curves of acetylcholine for inducing vascular relaxation shift rightward with a parallel manner, while the isomers IR-2alpha-2'R and IR-2alpha-2'S with a nonparallel manner. In addition, the antagonistic effects of the isomer IS-2alpha-2'R against the endothelial target for acetylcholine and against muscarinic receptors were 4570 and 10 times greater than those of the isomer IS-2alpha-2'S respectively. In conclusion, the endothelial target for acetylcholine had the unique pharmacological characteristics different from those of muscarinic receptors. PMID- 11883708 TI - Pharmacokinetics of ipriflavone, an isoflavone derivative, after intravenous and oral administration to rats hepatic and intestinal first-pass effects. AB - Pharmacokinetic parameters of ipriflavone were evaluated after intravenous administration of spray-dried ipriflavone with polyvinylpyrrolidone, SIP (5, 10, 20, and 40 mg/kg as ipriflavone) and oral administration of SIP (50, 100, and 200 mg/kg as ipriflavone) to rats. The hepatic, gastric, and intestinal first-pass effects of ipriflavone were also measured after intravenous, intraportal, intraduodenal, and oral administration of SIP (20 or 50 mg/kg as ipriflavone) to rats. After intravenous and oral administration, the pharmacokinetic parameters of ipriflavone were dose-independent. The extent of absolute oral bioavailability (F) was also independent of oral doses; the mean F value was approximately 24%. Considering the amount of unchanged ipriflavone recovered from 24-hr gastrointestinal tract (the mean value was approximately 12%), the low F values could be due to the hepatic, gastric, and/or intestinal first-pass effects. Based on total body clearance (CL) data of ipriflavone after intravenous administration, the first-pass effect in the heart and lung could be almost negligible, if any, in rats. Approximately 30% of ipriflavone absorbed into the portal vein was eliminated by liver (hepatic first-pass effect) based on intravenous and intraportal administration of SIP. The area under the plasma concentration-time curve from time zero to time infinity (AUC) values after oral administration and intraduodenal instillation of SIP, 50 mg/kg as ipriflavone, were not significantly different, but the values were significantly smaller (129 and 116 microg ml/min) than that after intraportal administration of SIP, 20 mg/kg as ipriflavone (513 microg ml/min based on 50 mg/kg), indicating that gastric first-pass effect of ipriflavone was negligible, but intestinal first pass effect was considerable in rats. Therefore, the low F value of ipriflavone after oral administration to rats was mainly due to intestinal first-pass effect. The hepatic first-pass effect and incomplete absorption of ipriflavone from rat gastrointestinal tract could also contributed to the low F in rats. PMID- 11883709 TI - Diazepam binding inhibitor (DBI) reduces testosterone and estradiol levels in vivo. AB - Diazepam binding inhibitor (DBI) is a putative endogenous ligand capable of binding to the central type benzodiazepine (BZD) receptor located on the GABAA receptor and the peripheral type BZD receptor on the mitochondrial outer membrane. We examined the effects of an intracerebroventricular injection of DBI on the serum levels of the gonadal hormones, testosterone and estradiol, respectively, in male and female mice. DBI (0.3-10 nmol/mouse, i.c.v.) significantly reduced the levels of both gonadal hormones in a dose-dependent manner. The decrease in the gonadal hormone levels became evident at 1 hr and lasted for at least 4 hrs after the DBI injection. The effects of DBI (3 nmol/mouse, i.c.v.) in male and female mice were completely attenuated by the coadministration of flumazenil (66 nmol/mouse), a selective antagonist for the central type BZD receptor. These results suggest that DBI acts as an endogenous modulator to regulate the levels of gonadal hormones in vivo, and that the DBI induced decrease in gonadal hormone levels is mediated by down regulation of the GABAergic system, implicated in gonadotropin-releasing systems and/or the hypothalamic-pituitary-gonadal axis. PMID- 11883711 TI - Effect of the organotin compound triethyltin on Ca2+ handling in human prostate cancer cells. AB - The effects of triethyltin on Ca2+ mobilization in human PC3 prostate cancer cells have been explored. Triethyltin increased [Ca2+]i at concentrations larger than 3 microM with an EC50 of 30 microM. Within 5 min, the [Ca2+]i signal was composed of a gradual rise and a sustained phase. The [Ca2+]i signal was reduced by half by removing extracellular Ca2+. The triethyltin-induced [Ca2+]i increases were inhibited by 40% by 10 microM nifedipine, nimodipine and nicardipine, but were not affected by 10 microM of verapamil or diltiazem. In Ca2+-free medium, pretreatment with thapsigargin (1 microM), an endoplasmic reticulum Ca+ pump inhibitor, reduced 200 microM triethyltin-induced Ca+ increases by 50%. Pretreatment with U73122 (2 microM) to inhibit phospholipase C did not alter 200 microM triethyltin-induced [Ca2+]i increases. Incubation with triethyltin at a concentration that did not increase [Ca2+]i (1 microM) in Ca2+-containing medium for 3 min potentiated ATP (10 microM)- or bradykinin (1 microLM)-induced [Ca2+]i increases by 41 +/- 3% and 51 +/- 2%, respectively. Collectively, this study shows that the environmental toxicant triethyltin altered Ca2+ handling in PC3 prostate cancer cells in a concentration-dependent manner: at higher concentrations it increased basal [Ca2+]i; and at lower concentrations it potentiated agonists-induced [Ca2+]i increases. PMID- 11883710 TI - Short term treatment with St. John's wort, hypericin or hyperforin fails to induce CYP450 isoforms in the Swiss Webster mouse. AB - This investigation was designed to determine whether St. John's wort (SJW)(435 mg/kg/d), a readily available antidepressant, or its purported active constituents hypericin (1 mg/kg/d) and hyperforin (10 mg/kg/d) were able to induce various hepatic cytochrome P450 (CYP450) isoforms. SJW, hypericin and hyperforin were administered to male Swiss Webster mice for four consecutive days and hepatic microsomes were prepared on day 5. None of the three treatments resulted in a statistical change in total hepatic CYP450 (SJW treated 0.95 +/- 0.09 nmol/mg vs control 1.09 +/- 0.14 nmol/mg). Furthermore, the catalytic activities of CYP1A2. CYP2E1 and CYP3A were unchanged from control following all three treatments as determined by ethoxyresorufin O-deethylation, p-nitrophenol hydroxylation and erythromycin N-demethylation respectively. Additionally, western immunoblotting demonstrated that there was no significant change in the polypeptide levels of any of the three isoforms. These results indicate that four days of treatment with moderate to high doses of SJW, hyperforin or hypericin fails to induce these CYP450 isoforms in the male Swiss Webster mouse. PMID- 11883712 TI - Trypsin is produced by and activates protease-activated receptor-2 in human cancer colon cells: evidence for new autocrine loop. AB - In this work, we showed that human colon cancer cell lines produce trypsin which can activate a receptor for trypsin, the protease-activated receptor-2 (PAR-2), in these cells. RT-PCR experiments showed that trypsinogen transcripts were present in four colon cancer cell lines: T84, Caco-2, HT-29 and C1.19A. By Western blot analysis we found a 25 kDa immunoreactive band identified as trypsinogen I in cell lysates and in the corresponding culture media. Concentrations of trypsin in cell media were found in nanomolar range, thus compatible with activation of protease-activated receptor 2 (PAR-2). This was further demonstrated in a colon cancer cell line (H-29) Ca2+i assay since increases in Ca2+i were observed in response to media from T84, Caco-2 or C1.19A cells that were similar to that observed with 2-5 nM trypsin and were abolished by trypsin inhibitor. Altogether, these data show that colon cancer cell lines produce and secrete trypsin at concentrations compatible with activation of PAR 2. They support possible autocrine/paracrine regulation of PAR-2 activity by trypsin in colon cancer cells. PMID- 11883713 TI - Anoxia-induced changes in purine nucleoside metabolism of in vitro aged human fibroblasts. AB - Inosine deriving from the metabolism of adenosine or inosine monophosphate (IMP) in the fibroblast provides the substrate for xanthine oxidase and is, therefore, an important source of toxic oxygen free radicals. With well-oxygenated medium, adenosine release appears to be greater for aged than young fibroblasts. In that the adenosine release by young cells is enhanced by reduced oxygenation, the effect anoxic stress on the release of the purine nucleosides adenosine and inosine by low-passage (PDL 23-26; young) vs. high-passage (PDL 43-51; aged) human lung fibroblasts (IMR-90) was studied. Cultures of confluent fibroblasts were incubated for 16 hr under normoxic (NF) or anoxic (AF) atmospheres. The release of adenosine and inosine was determined by HPLC at 0, 3, 6 and 24 hr after termination of the 16-hr period. Immediately following anoxia (time 0), adenosine release by young AF was 29% greater than for young NF, whereas both the youn PMID- 11883714 TI - Expression of swelling- and/or pH-regulated chloride channels (ClC-2, 3, 4 and 5) in human leukemic and normal immune cells. AB - Chloride channels on immune cells reportedly play important roles in cell volume regulation, cell proliferation and immune functions, but they are not well characterized at the molecular level. We examined the expression of swelling and/or pH-regulated chloride channels (ClC-2, 3, 4 and 5) in human leukemic cell lines [Jurkat and Hut-78 (T cells), Raji and Daudi (B cells), K-562 and HL-60 (myeloid cells)] and T cells, B cells and neutrophils from 8 normal subjects to clarify the difference of their expression among different cell types and maturity. Semi-quantitative RT-PCR and Northern blot analysis showed that ClC-3 was most abundantly expressed in all cells regardless of the cell types and maturity, while expression of ClC-2 was weak in these cells. Expression of ClC-4 was observed mainly in leukemic B cell lines, and in B cells and neutrophils from normal subjects. ClC-5 was expressed in all cell lines, while it was observed in only T and B cells but not in neutrophils from normal subjects. Thus, these chloride channels (ClC-2, 3, 4 and 5) showed distinct distribution among human immune cells, suggesting that they have specific roles in these cells. Molecular identification of chloride channels in leukocytes of different types and maturity may provide a new approach for the treatment of leukemia. PMID- 11883715 TI - Inhibition of placental ornithine decarboxylase by DL-alpha-difluoro-methyl ornithine causes fetal growth restriction in rat. AB - The roles of polyamines in intrauterine growth restriction (IUGR) is studied. The DL-alpha-difluoromethyl ornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase (ODC) which is a rate limiting enzyme of polyamine synthesis was administrated to pregnant rats so that we obtained rat fetuses with IUGR. The changes of maternal nutrition, damage of the placenta, and the direct effect of DFMO on the fetus were examined in this IUGR model. Administration of DFMO did not induced changes of maternal nutrition except for triglyceride and the fetal metabolic state. But the placental weight, ODC activity, and DNA in the placenta were decreased significantly. The ODC activity in the total placenta decreased to less than 10% of that of the control. Depression of ODC activity in the placenta may be the major cause of IUGR induced by DFMO administration, and polyamines play important roles to carry pregnancy. PMID- 11883716 TI - The hypothalamic levels of the endocannabinoid, anandamide, peak immediately before the onset of puberty in female rats. AB - Several data suggest that the endogenous cannabinoid system plays a role in neuroendocrine regulation in adult individuals, although the information on its involvement in peri-pubertal processes is scarce. In the present study, we have examined the ontogeny (from postnatal day [PND] 5 up to adulthood) of hypothalamic and anterior pituitary contents of anandamide (arachidonyl ethanolamide, AEA). We observed that the content of AEA in the hypothalamus was low at PND5, PND15 and PND25, but it markedly increased (approximately 3-fold) immediately before the puberty (on the day of 1st proestrus), to return to intermediate values immediately after the vaginal opening (day of 2nd proestrus) and, eventually, adulthood. By contrast, no consistent differences were observed in AEA levels in the anterior pituitary. These results demonstrate the occurrence of a parallelism between the peri-pubertal events and a rise in the hypothalamic content of AEA immediately before the puberty, which might indicate that this endocannabinoid may be involved in the onset of puberty in rats. PMID- 11883717 TI - Identification of cytokeratins as accessory mediators of Salmonella entry into eukaryotic cells. AB - Pathogenic Salmonella species initiate infection of a mammalian host by inducing their own uptake into intestinal M-cells. During the uptake process, the bacteria utilize an intrinsic secretion system to release proteins that enter host cells. The secreted invasion-mediating proteins subsequently interact with host cell components that induce alterations in the actin cytoskeleton. To identify potential cellular determinants of invasion, we employed a yeast two-hybrid system using the secreted Salmonella invasion protein (SipC) as the bait protein. This system identified cytokeratins, supportive components of the cytoskeletal matrix, as proteins that may physically interact with SipC. Transfection-based studies revealed an inhibition of Salmonella invasion when a dominant negative cytokeratin-18 was expressed. Immunofluorescent confocal microscopy studies revealed that Salmonella did not enter HEp-2 cells expressing the dominant negative cytokeratin-18. These results suggest that an interaction between SipC and cytokeratin-18 may occur as part of Salmonella invasion. PMID- 11883718 TI - Deoxamuscaroneoxime derivatives as useful muscarinic agonists to explore the muscarinic subsite: demox, a modulator of orthosteric and allosteric sites at cardiac muscarinic M2 receptors. AB - A series of muscarinic agonists, straight chained, branched, cyclic alkyl and aromatic derivatives of the oxime 1 (demox) was designed with the aim of investigating their activity on muscarinic receptor subtypes. Effects on M1 receptor were assessed functionally by a microphysiometer apparatus, while M2, M3, and M4 receptor potency and affinity were studied on isolated preparations of guinea pig heart, ileum, and lung, respectively. The results suggest that the substitution of a hydrogen with a long side-chain or bulky group generally induces a decrease in potency at M1 and M3 subtypes, while a general increase in this parameter is obtained at M2 subtype. Among the agonists 2-18, compound 4 behaves as a full agonist with a preference for M3 subtype. Moreover, compound 12 is inactive at M1 and M4 receptors while it displays a full agonist activity at M2 and M3 subtypes. Since demox displays a variable response on cardiac M2 receptors regulating heart force, an in-depth inquiry of the functional behaviour of this compound was carried out at M2 receptors. In presence of 10(-11) and 10( 10) M demox, the binding of [3H]-NMS was increased by approximately 30% as a consequence of an increase of the association of [3H]-NMS to membranes; this effect was not observed in presence of a higher concentration of [3H]-NMS. Higher concentrations of demox decreased the binding of [3H]-NMS to heart atrial membranes but significantly retarded the dissociation of this radioligand. Our results suggest that demox may interact with orthosteric and allosteric sites of atrial M2 muscarinic receptor. PMID- 11883719 TI - Silybin and its bioavailable phospholipid complex (IdB 1016) potentiate in vitro and in vivo the activity of cisplatin. AB - In this study we investigated whether the flavonoid silybin and its bioavailable derivative IdB 1016 (silipide) could enhance the antitumour activity of cisplatin (CDDP), the most commonly used drug in the treatment of gynaecological malignancies. Silybin alone up to 10 (M was unable to produce a relevant in vitro growth inhibition of A2780 cells, whereas CDDP was effective, giving an IC50 value of 0.5+/-0.14 microM. When silybin was combined with CDDP, a dose-dependent and statistically significant (p<0.05) increase of the CDDP activity was noticed, yielding IC50 values of 0.35+/-0.07 and 0.263+/-0.004 microM at silybin concentrations of 1 and 10 microM, respectively. The same trend was observed for in vivo experiments. IdB 1016 alone (1350 mg/kg) did not significantly affect tumour growth, whereas CDDP at the Maximum Tolerated Dose (12 mg/kg) produced a tumour weight inhibition (TWI%) of 80% and a log10 cell kill (LCK) of 0.7. Administration of both drugs resulted in a potentiation of the antitumour activity and TWI% and LCK increased to 90% and 1, respectively. Interestingly, mice receiving the combination recovered earlier in terms of body weight loss as compared to CDDP-treated mice. CDDP at 6 mg/kg yielded TWI of 44% and LCK of 0. The concomitant administration of IdB 1016 (1800 mg/kg) enhanced CDDP anti-tumour activity, with 68% TWI and 0.6 LCK. Finally, an antiangiogenic effect of IdB 1016 in an in vivo experimental model was demonstrated. Median haemoglobin value for the Matrigel from the vehicle-treated controls was 2.43 versus a value of 0.321 for the IdB 1016-treated animals. PMID- 11883720 TI - Enhanced secretion of tissue plasminogen activator by simultaneous use of retinoic acid and ascorbic acid from tissue cultured gastroepiploic artery. AB - Tissue-type plasminogen activator (tPA) is a key enzyme in the fibrinolysis system and the regulation of its expression has been extensively studied in cultured vascular endothelial cells. Many kinds of supplements including growth factors are needed, however, to keep endothelial cells viable, which leads the culture condition far from the physiological milieu. Using a new device of amorphous calcium phosphate coated culture plate, we succeeded in culturing ring cut gastroepiploic artery in a basic medium of RPMI 1640 containing 10% fetal calf serum. The overall normal vessel architecture and the antigenicity of von Willebrand factor, tPA and plasminogen activator inhibitor type 1 (PAI-1) were retained for at least 9 days. tPA was constantly secreted into the conditioned medium at least up to day 12. Employing this organ culture technique, we analyzed the effects of two well-known profibrinolytic vitamins of retinoic acid (Vit. A) and ascorbic acid (Vit. C) on the release of tPA and PAI-1. The cultured artery responded well and the tPA secretion was enhanced by factors of 1.5 fold by Vit. A, 1.7 fold by Vit C and 3.2 fold by their combination, whereas none of these stimuli increased PAI-1 secretion. These results suggested that the cultured ring cut artery retained functional endothelial cells for at least 9 days and was suitable in analyzing the regulatory mechanism of protein synthesis and secretion from the vascular wall. Using this method, vitamins A and C were shown to lead the intravascular condition to a profibrinolytic state. PMID- 11883721 TI - Fatigue, depression and chronic hepatitis C infection. AB - BACKGROUND: We aimed to determine if an association exists between uncomplicated hepatitis C virus (HCV) infection and depression or fatigue. METHOD: A review of the literature was undertaken. RESULTS: There is an association between HCV infection and either depression or fatigue in certain circumstances--those who are aware they are HCV positive, those with advanced liver disease and those seen in specialist referral centres. All these studies are subject to important biases. There are only a few studies in which knowledge of HCV status and assessment of fatigue or depression is independent. These studies do not suggest an association. There is no association between conventional markers of liver disease and depression or fatigue. CONCLUSIONS: Despite anecdotal evidence to the contrary, at the moment there is no evidence that HCV infection per se is associated with fatigue or depression, and there is a suggestion that it is not. The same risk factors that exist for fatigue in other physical illnesses, such as metabolic disorder, mood disorder, demographics and lack of exercise, certainly exist for HCV. Although there are elegant theoretical mechanisms, there is no compelling epidemiological evidence for an additional HCV specific fatigue or depression factor. PMID- 11883723 TI - Prevalence, incidence and stability of premenstrual dysphoric disorder in the community. AB - BACKGROUND: Despite an abundance of clinical research on premenstrual and menstrual symptoms, few epidemiological data provide estimates of the prevalence, incidence, co-morbidity, stability and correlates of premenstrual dysphoric disorder (PMDD) in the community. AIMS: To describe the prevalence, incidence, 12 co-morbidity factors and correlates of threshold and subthreshold PMDD in a community sample of young women. METHODS: Findings are based on prospective longitudinal community survey of 1488 women aged 14-24, who were followed-up over a period of 48 months (follow-up, N = 1,251) as part of the EDSP sample. Diagnostic assessments were based on the Composite International Diagnostic Interview (CIDI) and its 12-month PMDD diagnostic module administered by clinical interviewers. Diagnoses were calculated using DSM-IV algorithms, but daily ratings of symptoms, as required, were not available. RESULTS: The baseline 12 month prevalence of DSM-IV PMDD was 5.8%. Application of the diagnostic exclusion rules with regard to concurrent major depression and dysthymia decreased the rate only slightly (5.3%). An additional 18.6 % were 'near-threshold' cases, mostly because they failed to meet the mandatory impairment criterion. Over the follow up period only few new PMDD cases were observed: cumulative lifetime incidence was 7.4%. PMDD syndrome was stable across 48 months with < 10% complete remissions among baseline PMDD cases. The 12-month and lifetime co-morbidity rates were high (anxiety disorders 47.4%, mood disorders 22.9%; somatoform 28.4%), only 26.5 % had no other mental disorder. Particularly high odds ratios were found with nicotine dependence and PTSD. In terms of correlates increased rates of 4-weeks impairment days, high use of general health and mental health services, and increased rates of suicide attempts were found. CONCLUSION: In this sample of adolescents and young adults, premenstrual symptoms were widespread. However, DSM-IV PMDD was considerably less prevalent. PMDD is a relatively stable and impairing condition, with high rates of health service utilization, increased suicidality and substantial co-morbidity. PMID- 11883722 TI - A twin study of genetic and environmental influences on suicidality in men. AB - BACKGROUND: Previous studies that have examined genetic influences on suicidal behaviour were confounded by genetic vulnerability for psychiatric risk factors. The present study examines genetic influences on suicidality (i.e. suicidal ideation and/or suicide attempt) after controlling for the inheritance of psychiatric disorders. METHODS: Sociodemographics, combat exposure, lifetime DSM III-R major depression, bipolar disorder, childhood conduct disorder, adult antisocial personality disorder, panic disorder, post-traumatic stress disorder, drug dependence, alcohol dependence and lifetime suicidal ideation and attempt were assessed in 3372 twin pairs from the Vietnam Era Twin Registry who were assessed in 1987 and 1992. Genetic risk factors for suicidality were examined in a multinomial logistic regression model. Additive genetic, shared environmental and non-shared environmental effects on suicidality were estimated using structural equation modelling, controlling for other risk factors. RESULTS: The prevalence of suicidal ideation and suicide attempt were 16.1% and 2.4% respectively. In a multinomial regression model, co-twin's suicidality, being white, unemployment, being other than married, medium combat exposure and psychiatric disorders were significant predictors for suicidal ideation. Co twin's suicidality, unemployment, marital disruption, low education attainment and psychiatric disorders (except childhood conduct disorder) were significant predictors for suicide attempt. Model-fitting suggested that suicidal ideation was influenced by additive genetic (36%) and non-shared environmental (64%) effects, while suicide attempt was affected by additive genetic (17%), shared environmental (19%) and non-shared environmental (64%) effects. CONCLUSIONS: There may be a genetic susceptibility specific to both suicidal ideation and suicide attempt in men, which is not explained by the inheritance of common psychiatric disorders. PMID- 11883724 TI - Stress and well-being in mothers of young children 11 years after the Chornobyl nuclear power plant accident. AB - BACKGROUND: This paper examines the association between exposure to the Chornobyl nuclear power plant explosion and the psychological and physical well-being of mothers with young children. The study also examines whether exposure to Chornobyl increased the vulnerability of mothers to subsequent economic and social stress, and thus represents a unique test of the stress-vulnerability model in a non-Western setting. METHOD: The sample consisted of mothers evacuated from the contamination zone surrounding the plant (evacuees) and mothers who had never lived in a radiation-contaminated area (controls). In addition to exposure status, the interview obtained data on perceived economic stress, social stress and stress moderators. The dependent variables were measured by the SCL-90 global severity index (GSI), perceived physical health and number of days unable to work due to illness. RESULTS: Overall, evacuees reported fewer stressors and greater personal and social resources than control mothers. Nevertheless, evacuees scored higher on the GSI, reported lower perceived physical health and took more sick days relative to control mothers, even after controlling for demographic factors, stressors and stress moderators. Tests of interaction effects were not statistically significant. CONCLUSIONS: The findings confirmed that married women with young children evacuated to Kyiv following the Chornobyl nuclear power plant explosion reported significantly poorer psychological and perceived physical health than controls 11 years later. Although perceived social and economic adversities also affected these outcomes, there was no evidence that exposure to the Chornobyl accident increased the vulnerability of mothers to these stressors, giving support to the additive burden model of stress. PMID- 11883725 TI - Beliefs, sense of control and treatment outcome in post-traumatic stress disorder. AB - BACKGROUND: Few studies have shown that maladaptive beliefs relate to treatment outcome. METHOD: In a randomized controlled study, 87 patients with post traumatic stress disorder (PTSD) had exposure therapy alone or cognitive restructuring alone, or both combined, or relaxation. Independent blind assessors assessed patients at pre-, mid- and post-treatment and at follow-up; at those times patients rated cognitive, behavioural and emotional aspects of their disorder. RESULTS: Baseline beliefs about mistrust, helplessness, meaninglessness and unjustness of the world related to baseline PTSD symptoms but did not predict treatment outcome, though improvement in certain beliefs correlated with more symptom improvement. Several 'key' beliefs changed after, and none before, symptoms improved. At post-treatment, sense of control and attribution of gains to personal efforts predicted maintenance of gains at follow-up. CONCLUSIONS: Baseline beliefs and improvement in beliefs did not predict outcome. Post treatment sense of control/internal attribution predicted maintenance of gains at follow-up. How much sense of control is produced by or causes improvement deserves testing. PMID- 11883726 TI - Childhood behaviour, psychotic symptoms and psychosis onset in young people at high risk of schizophrenia: early findings from the edinburgh high risk study. AB - BACKGROUND: Several studies suggest that many patients with schizophrenia have pre-morbid neurodevelopmental abnormalities. This study examines how behavioural abnormalities are associated with mild psychotic symptoms and later schizophrenic illness. METHODS: Maternal ratings on the Child Behavior Checklist (CBCL) of the early behaviour of 155 subjects were obtained at entry to the Edinburgh study of people at high risk of schizophrenia. These maternal ratings were compared in those with and without psychotic symptoms and used to predict the later onset of psychosis. RESULTS: The CBCL syndrome scores for the children prior to age 13 did not distinguish any of the study groups at entry to the study. In the ratings made for the subjects when aged from 13 to 16, delinquent behaviour and 'other problems' were weakly associated with these symptoms. However, with the exception of somatic symptoms and thought problems, the age 13-16 scales were significant predictors of later schizophrenic illness. This was true also for some of the ratings prior to age 13. CONCLUSIONS: Various behaviours, in particular, withdrawn and delinquent-aggressive behaviour in adolescents at risk of schizophrenia may predict later onset of the illness. These behaviours, however, are far less predictive of isolated psychotic symptoms prior to psychosis onset. PMID- 11883727 TI - Siblings of schizophrenia patients could be distinguished from siblings of bipolar patients. PMID- 11883728 TI - Genetic and environmental influences on premenstrual symptoms in an Australian twin sample. AB - BACKGROUND: We aimed to explore the prevalence and factor structure of premenstrual symptoms in a sample of Australian twins; to investigate phenotypic associations between reported premenstrual symptoms, personality and reproductive dimensions; and to identify the relative contributions of genes and environment to premenstrual symptoms and the extent of genetic and environmental covariation with the personality trait Neuroticism and lifetime major depression. METHOD: Seven hundred and twenty female twin pairs (454 monozygotic and 266 dizygotic) from the Australian National Health and Medical Research Council Twin Register reported on experience of 17 premenstrual symptoms during the previous 12 months. In the same questionnaire twins also responded to questions on symptom states, and personality dimensions including neuroticism. Interview data enabling diagnosis of lifetime history of DSM-IV major depression were also available. We fitted univariate and multivariate genetic models to the data. RESULTS: Most frequently reported symptoms were breast tenderness/pain and bloating/weight gain, followed by affective symptoms. Twelve-month prevalence was 24% for the combination of symptoms and functional interference meeting a very rough approximation of DSM-III-R criteria for late luteal dysphoric disorder. Principal factor analysis identified a single premenstrual (PMS) factor. Additive genetic influences (44% of total variance) were identified for PMS. Although we found genetic correlations of 0.62 between reported PMS and neuroticism, and 0 70 with lifetime major depression, 39 % of the genetic variance of PMS was not explained by these factors. CONCLUSIONS: Our findings support the existence of genetic influences on premenstrual symptoms, but we were unable to distinguish between liability to symptom experience and symptom reporting. Retrospective reporting may have contributed to our finding that PMS genes were shared in part with neuroticism and liability to lifetime major depression. PMID- 11883729 TI - Genetic effects on the variation and covariation of attention deficit hyperactivity disorder (ADHD) and oppositional-defiant disorder/conduct disorder (Odd/CD) symptomatologies across informant and occasion of measurement. AB - BACKGROUND: Previous studies have shown that the presence of conduct disorder may contribute to the persistence of attention deficit-hyperactivity disorder (ADHD) symptomatology into adolescence; however, the aetiological relationship between the two phenotypes remains undetermined. Furthermore, studies utilizing multiple informants have indicated that teacher ratings of these phenotypes are more valid than maternal reports. METHODS: The genetic structure underlying the persistence of ADHD and oppositional-defiant disorder/conduct disorder (ODD/CD) symptomatologies as rated by mothers and teachers at two occasions of measurement was investigated on a sample of 494 male and 603 female same sex adolescent twin pairs participating in the Virginia Twin Study of Adolescent Behavioral Development (VTSABD). RESULTS: Using structural modelling techniques, one common genetic factor was shown to govern the covariation between the phenotypes across informants and occasion of measurement with additional genetic factors specific to ODD/CD symptomatology and persistence of symptomatology at reassessment. Genetic structures underlying the phenotypes were, to some extent, informant dependent. CONCLUSIONS: The findings indicate that it is unlikely that the co morbidity between ADHD and ODD/CD is due to environmental influences that are independent of ADHD. Rather it is likely to be due to a shared genetic liability either operating directly, or indirectly through gene-environment correlations or interactions. The covariation between phenotypes across informants and time is governed by a common set of genes, but it seems that ODD/CD is also influenced by additional genetic factors. Developmentally, different forms of genetic liability control ADHD in males and inattention in females. PMID- 11883730 TI - Sibling pairs with affective disorders: resemblance of demographic and clinical features. AB - BACKGROUND: As part of a collaborative linkage study, the authors obtained clinical and demographic data on 160 families in which more than one sibling was affected with a bipolar illness. The aim of the study was to identify clinical characteristics that had a high degree of familiality. METHOD: Data on age at onset, gender, frequency of illness-episodes and proportion of manic to depressive episodes were examined to determine intra-pair correlations in affected sibling pairs. Dimension scales were developed measuring frequency and severity of lifetime mania, depression, psychosis and mood-incongruence of psychotic symptoms; degree of familial aggregation for scores on these dimensions was calculated. RESULTS: Sibling pairs correlated significantly for age at onset (p = 0.293, P < 0 001); dimension scores for psychosis (p = 0.332, P < 0.001); and proportion of manic to depressive episodes (p = 0.184, P = 0.002). These findings remained significant when correcting for multiple testing. Of the other test variables; mania (p = 0.171, P = 0.019); incongruence dimensions (p = 0.242, P = 0.042); .frequency of manic episodes (p = 0.152, P = 0.033); and frequency of depressive episodes (p = 0.155, P = 0.028) were associated with modest correlations but these were not significant after correction. Degree of familial aggregation was not significant for sex (kappa = 0.084) or dimension scores for depression (p = 0.078, P = 0.300). CONCLUSIONS: Significant but modest familial resemblance has been shown for some specific features of bipolar illness, particularly age at onset and degree of psychosis. Further research may establish the extent to which these findings are mediated by genetic and/or environmental factors. PMID- 11883731 TI - Parental alcohol use disorders and alcohol use and disorders in offspring: a community study. AB - BACKGROUND: We examined the association between parental alcohol use disorders and patterns of alcohol consumption and DSM-IV alcohol use disorders in their offspring in a community-based sample of young adults. METHODS: Data are based on baseline and 4-year follow-up data of 2427 respondents aged 14-24 at baseline. Alcohol use and disorders in respondents were assessed using the Munich-Composite international-Diagnostic-Interview with DSM-IV algorithms. Diagnostic information about parents was collected by family history information from the respondents, and by direct interview with one parent (cohort aged 14 to 17 years only). RESULTS: Although the association between maternal and paternal alcohol use disorders and non-problematical drinking in offspring was minimal, there was a strong effect for the transition to hazardous use and for alcohol abuse and dependence; the effect of parental concordance for transition into hazardous use was particularly striking. Maternal history was associated with a higher probability of progression from occasional to regular use, whereas paternal history was associated with progression from regular to hazardous use. Parental alcoholism increased the risk for first onset of hazardous use and alcohol dependence between the ages of 14-17, and for an earlier onset of the alcohol outcomes in offspring. The impact of parental alcohol use disorders was comparable for male and female offspring. CONCLUSIONS: Parental alcoholism predicts escalation of alcohol use, development of alcohol use disorders and onset of alcohol outcomes in offspring. PMID- 11883732 TI - Early adolescent marijuana use: risks for the transition to young adulthood. AB - BACKGROUND: This study assessed the relationship of early adolescent marijuana use to performance of developmental tasks integral to the transition to young adulthood. The tasks concerned intimacy, education, and work and social conformity. METHODS: African American (N = 617) and Puerto Rican (N = 531) youths completed questionnaires in their classrooms. Five years later they were individually interviewed. Logistic regression analysis estimated the increased likelihood that early marijuana users would make an inadequate transition to young adult social roles. RESULTS: Analyses examining the association between early marijuana use and 20 outcome variables found significant relationships for 10 of them: (a) having lower educational and occupational expectations; (b) being suspended or expelled from school, fired from jobs, 'high' at school or work, collecting welfare; and (c) rebelliousness, not participating in productive activities, not attending church, and being an unmarried parent. Marijuana use was not related to any of the intimate relationship measures. These finding emerged with controls on gender, ethnicity, age and mother's education. CONCLUSIONS: Among African Americans and Puerto Ricans, early marijuana use predicts less adequate performance on some developmental tasks integral to becoming an independent young adult. Marijuana is not a benign drug and is associated with future risks for the individual and society at large. PMID- 11883734 TI - Occupational asthma. PMID- 11883733 TI - Circumscribed numerical deficit of dorsal raphe neurons in mood disorders. AB - BACKGROUND: Neurocircuits comprising limbic, striato-pallidal and thalamo cortical brain areas are assumed to be involved in the pathophysiology of mood disorders. All these brain regions receive serotonergic afferents arising from the rostral raphe, mainly the dorsal raphe. Although serotonergic systems appear to be involved in the pathology of mood disorders, there is uncertainty as to whether structural alterations in raphe nuclei exist alongside a functional dysregulation of the serotonergic system. METHODS: In the brains of 12 patients with mood disorders (major depressive disorder N= 6, bipolar disorder N = 6) and 12 normal subjects we performed a morphometric post-mortem study on neuronal morphology in all subnuclei of the dorsal raphe nucleus using Nissl stained 20 microm axial serial sections of the brainstem. RESULTS: The number of neurones of the ventrolateral subnucleus of the dorsal raphe was reduced by 31 % in patients with mood disorders compared with non-psychiatric control subjects. Ventrally located subnuclei of the rostral dorsal raphe (ventrolateral, ventral, interfascicular) taken together also showed a smaller number of neurones. Neurone numbers of the dorsal and the caudal subnucleus and volumes of all single subnuclei appeared to be unchanged. Analysis of morphological neuronal types revealed a smaller number of triangular neurones in the ventrolateral subnucleus. Numbers of ovoid and round neurones in the ventrolateral subnucleus also showed a trend to reduction. No correlation was found between neurone numbers in any subnucleus of the dorsal raphe and duration of illness. Neurone numbers did not differ in any subnucleus between patients with unipolar and those with bipolar affective disorder. CONCLUSIONS: Results indicate that patients with primary mood disorders have a circumscribed numerical neuronal deficiency in the dorsal raphe. This structural deviation may contribute to impaired serotonergic innervation of brain regions which are involved in the pathology of mood disorders. PMID- 11883735 TI - The antiinflammatory activity of budesonide on human airway epithelial cells is lasting after removal of the drug from cultures. AB - Because of its ability to conjugate extensively with fatty acids within lung cells, it has been suggested that budesonide (Bud) may have a prolonged pharmacologic activity, related to retention of the drug in airway tissues. Using human bronchial epithelial cells (HBECs) as target cells, we evaluated whether Bud could have a long-lasting inhibitory effect on ICAM-1 expression and GM-CSF release. HBECs were cultured in Bud (10 microM) or in medium alone (Ctr) for 24 hr, then extensively washed (to remove Bud) and incubated for an additional 6, 12, or 24 hr with IFN-gamma. ICAM-1 expression and GM-CSF release were then measured by flow cytometric analysis. In Ctr HBECs, IFN-gamma induced a time dependent upregulation of ICAM-1 expression, significant at 6, 12, or 24 hr (p < 0.05, each comparison), and an increase in GM-CSF release, significant at 24 hr (p < 0.05). The inhibitory effects of Bud preexposure on IFN-gamma-induced ICAM-1 expression and GM-CSF release were then compared with those of a continuous exposure to the drug during IFN-gamma stimulation. Preexposure to Bud (1 and 10 microM) induced a significant inhibition of IFN-gamma-induced ICAM-1 expression (p < 0.05, each comparison), but lower than that observed in HBECs continuously exposed at the same Bud concentrations (p < 0.01, each comparison). In contrast, the inhibition of GM-CSF release was similar in preexposed and in exposed HBECs and statistically significant only at the highest Bud concentration tested (p < 0.05, each comparison). Thus, Bud is effective in vitro in inducing a downregulation lasting 24 hr of mechanisms involved in leukocyte recruitment. PMID- 11883736 TI - Synergistic effect of theophylline and procaterol on interleukin-5-induced degranulation from human eosinophils. AB - Inhibiting the release of toxic granule proteins from eosinophils is a possible means of treating allergic inflammation. This study was performed to examine whether procaterol and theophylline, commonly used bronchodilators in asthma, inhibit eosinophil degranulation induced by interleukin (IL)-5. Purified eosinophils from patients with asthma were incubated with IL-5 for 24 hr in the presence of theophylline, procaterol, combinations of theophylline and procaterol, or dexamethasone. Levels of eosinophil-derived neurotoxin (EDN) in the supernatants were measured with radioimmunoassay. Theophylline inhibited IL-5 induced release of EDN in a concentration-dependent manner. Procaterol inhibited degranulation only at high concentrations. However, procaterol at 10(-9) M and 10(-8) M, which are physiologic concentrations, together with theophylline at 10( 5) M, which is a concentration commonly found in the serum of patients receiving low-dose theophylline, inhibited degranulation by 43.8%. This finding indicates that theophylline and procaterol have synergistic effects. The inhibition was comparable to that with dexamethasone at 10(-9) M. Our results suggest that a combination of low-dose theophylline and procaterol exhibits antiinflammatory effects in asthma by inhibiting eosinophil-effector functions. PMID- 11883737 TI - A multi-stage asthma screening procedure for elementary school children. AB - This paper describes an asthma screening procedure developed to identify children with asthma for an intervention study. Students were classified into three categories based on questionnaire responses (previous asthma, suspected asthma, and no evidence of asthma). Those classified as suspected asthma by questionnaire underwent further testing, including spirometry and exercise challenge. Using the questionnaire alone, the measured asthma prevalence was 32%; the addition of spirometry and step testing reduced this estimate to 9.89%. The diagnosis of asthma was confirmed in 96% of children who saw the study physician. This screening procedure can identify school children with suspected undiagnosed asthma. PMID- 11883738 TI - Which factors predict success after discontinuation of inhaled budesonide therapy in children with asthma? AB - Urinary eosinophil protein X (UEPX) concentration, lung function, and nonspecific bronchial hyperreactivity were determined in 40 asthmatic children (asymptomatic for 6.4 +/- 3.0 months) (mean age 9.8 +/- 2.9 years) receiving inhaled budesonide, in order to establish whether measurement of these parameters is useful in determining discontinuation of inhaled corticosteroid therapy. After the discontinuation of therapy, patients were asked to come to the Outpatient Clinic if symptoms recurred and did not respond to beta2 mimetic usage in 24 hr. Otherwise they were to be seen 2-3 months later for a follow-up visit. UEPX concentration was determined and spirometry was performed on this visit. While UEPX concentrations had increased (p < 0.0001), FEV1, FEF 25-75 and PEF had decreased significantly 2.3 +/- 0.53 months after the cessation of inhaled budesonide therapy in all children (p = 0.004, p = 0.02, p = 0.02, respectively). Due to clinical deterioration, inhaled corticosteroid therapy had to be restarted in 19 (48%) of the children (Group I), while the remaining 21 (52%) (Group II) continued to be asymptomatic during the 2.3 +/- 0.5 months follow-up period. Although the initial UEPX concentrations, spirometer variables, and methacholine PC20 values of these two groups were not statistically different, the duration of clinical remission before discontinuation of budesonide prophylaxis was significantly longer in group II (p = 0.0037). We concluded that, in determining discontinuation of inhaled corticosteroid prophylaxis, duration of clinical remission seems to be a more useful criterion than measurement of UEPX levels, lung function test, and assessment of bronchial hyperreactivity. PMID- 11883739 TI - Enhancing medication adherence among inner-city children with asthma: results from pilot studies. AB - Despite the availability of effective treatments that aid in controlling asthma symptoms, inner-city children with asthma have high rates of morbidity and are frequent users of emergency department services. The goal of these studies was to pilot test an intervention that used social learning strategies (e.g., goal setting, monitoring, feedback, reinforcement, and enhanced self-efficacy) and targeted known barriers to individualize a family-based asthma action plan. Participants were 15 children with asthma, aged 7-12 years, who had been prescribed at least one daily inhaled steroid. The children and their mothers lived in inner-city Baltimore and all were African-American. Participants received up to five visits in their home by a nurse. Electronic monitors were installed on the children's MDI to provide immediate feedback on medication adherence to the families and validate medication use. At baseline, only 28.6% of the children were using their medications as prescribed. Within four weeks, the number of children who were using their medications appropriately doubled from 28.6% at baseline to 54.1% (90% increase; p = 0.004), while underutilization decreased from 51.2% to 25.4% (100% decrease; p = 0.02). The number of children with no medication use at all dropped from 28.3% at baseline to 15.1% by week 5 (87% decrease; p = 0.009). Thus, within four weeks, more than half the children were using their inhaled steroids appropriately. In addition, the rate of underutilization decreased and that of nonutilization was cut in half. Our initial data suggest that an individualized, home-based intervention can significantly enhance adherence to the daily use of inhaled steroids in inner city children with asthma. Nevertheless, adherence to daily inhaled steroid therapy remains a significant problem in this group. PMID- 11883740 TI - Using case management to increase antiinflammatory medication use among a managed care population with asthma. AB - The National Heart, Lung, and Blood Institute (NHLBI) recommends the regular use of antiinflammatory medications to achieve and maintain control of persistent asthma, while recommending that quick-relief beta2-agonist medications should be used to treat acute symptoms and exacerbations. Despite these suggestions, the overuse of short-acting quick-relief medications and underuse of long-acting antiinflammatory medications persists. ConnectiCare, Inc., a regional managed care company, used pharmacy claims data to identify members who had been dispensed a total of three or more beta2-agonist prescriptions for three consecutive months in a 12-month period. These members had also not been prescribed an inhaled corticosteroid, cromolyn sodium, or nedocromil during the same three consecutive months. An intensive case management intervention was developed that included multiple contacts from a nurse case manager to provide education and information about asthma control and the guidelines. Twenty-eight percent (n = 40) were purposively chosen to receive the intensive multiple contact intervention and the remainder received a standard, single-contact intervention. After adjusting for the effects of age, gender, and pre intervention medication use, ConnectiCare members who received the intensive intervention were 4.3 times more likely to increase the number of antiinflammatory medication prescriptions dispensed that those who received a standard intervention (p < 0.001). This study suggests that the use of intensive case management for persons identified as inappropriate users of asthma medication may result in medication changes that achieve the long-term control of asthma. PMID- 11883742 TI - Efficacy of inhaled anticholinergics and anesthesia in treatment of a patient in status asthmaticus. PMID- 11883741 TI - Once-daily budesonide inhalation powder (Pulmicort Turbuhaler) is effective and safe in adults previously treated with inhaled corticosteroids. AB - This 12-week, double-blind, placebo-controlled, multicenter study evaluated the efficacy and safety of budesonide 400 microg administered once daily via Turbuhaler in adults previously treated with at least twice-daily dosing of inhaled corticosteroids. Pulmonary function (FEV1, PEF, FVC, FEF(25-75%)), asthma symptom scores, quality of life, and breakthrough medication use were significantly (p < 0.05) different in patients receiving once-daily budesonide Turbuhaler compared to placebo, and significantly (p < 0.001) more patients receiving placebo discontinued the study. Adverse events were similar between study groups. Once-daily administration of budesonide Turbuhaler was safe and efficacious in patients previously treated with inhaled corticosteroids. PMID- 11883743 TI - Genetic contribution of the interleukin-10 promoter polymorphism in endometriosis susceptibility. AB - PROBLEM: Interleukin-10 (IL-10) is an important immunomodulatory cytokine. The aim of this study was to investigate whether polymorphisms of the IL-10 gene promoter polymorphisms may be responsible in part for genetic susceptibility to endometriosis. METHODS OF STUDY: Polymorphisms at position -1082 and -592 in the IL-10 promoter region were determined in 196 patients with endometriosis and 160 fertile healthy women by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). RESULTS: There were no statistically significant differences in the genotype and allele frequencies of IL-10 promoter polymorphism between the endometriosis and control groups. However, when subgrouped according to clinical features, the frequencies of the -592*CC genotype and -592*C allele were significantly increased in patients with autoantibodies to carbonic anhydrase II (anti-CA II ab) compared with controls. CONCLUSION: IL-10 promoter polymorphisms were associated with the production of anti-CA II ab in patients with endometriosis, suggesting a role in the genetic susceptibility for endometriosis. PMID- 11883744 TI - Increased levels of macrophage colony-stimulating factor in the placenta and blood in preeclampsia. AB - PROBLEM: Macrophage colony-stimulating factor (M-CSF) is considered an essential cytokine for placental growth and maintenance. We evaluated whether M-CSF levels in the placenta and blood in preeclampsia differed from those in normal pregnancies. METHOD OF STUDY: The subjects were 37 pregnant women carrying single fetuses, of whom 19 were women with normal pregnancies and 18 were women with preeclampsia. Their average gestational age at entry was 38 weeks of gestation. Blood was collected before the onset of labor, and separated serum was obtained after centrifugation. A tissue segment of the placenta was cut immediately after delivery. The frozen placental tissue was placed in a plastic tube containing phosphate-buffered saline. The tissue was fully homogenized and then centrifuged. Separated supernatant was used for subsequent determination. M-CSF levels in separated serum were measured, and M-CSF and total protein (TP) levels in separated supernatant were also measured. RESULTS: Both M-CSF/TP levels in the placenta and M-CSF levels in blood were significantly higher (P < 0.05-0.01) in preeclampsia than in normal pregnancies. CONCLUSIONS: This is the first report concerning high placenta levels of M-CSF/ TP in preeclampsia. Increased M-CSF in the placenta supports the hypothesis that immunological abnormalities contribute to the etiology of preeclampsia. PMID- 11883745 TI - Expression of pro-inflammatory cytokines in mouse blastocysts during implantation: modulation by steroid hormones. AB - PROBLEM: Expression and hormonal regulation of pro-inflammatory cytokines and their role in blastocyst activation and implantation is poorly known. The present study is aimed at analysing the expression and hormonal modulation of two pro inflammatory cytokines [interleukin-1alpha (IL-1alpha) and IL-6] in mouse blastocysts during implantation. METHOD OF STUDY: Blastocyst-uterine interactions are inhibited by progesterone during implantation and subsequent treatment with oestrogen triggers events that allow implantation to begin. Using this delayed implantation mouse model, dormant and activated blastocysts were recovered from mice treated with progesterone alone and progesterone plus oestrogen therapy, respectively. Expression of IL-1alpha and IL-6 messenger RNA (mRNA) was analysed in normal, dormant and activated blastocysts by in situ hybridization using specific labelled sense and antisense RNA probes, and the protein expression of the same was analysed by immunocytochemistry. RESULTS: In situ hybridization revealed IL-1alpha and IL-6 mRNA localization in normal, dormant and activated blastocysts and a differential expression was observed in relation to the exposure to progesterone and oestrogen. There was less expression in the dormant blastocysts as compared with the normal and activated ones, and the pattern was similar for both cytokines. Immunocytochemistry also revealed a similar pattern of protein expression to that of the mRNA expression for both the cytokines. CONCLUSIONS: Using a delayed implantation model, we show that mouse blastocysts express both IL-1alpha and IL-6 mRNA as well as their respective proteins. Both mRNA and the protein levels of IL-1alpha and IL-6 seem to be hormonally modulated in mouse blastocysts during implantation. PMID- 11883746 TI - Dynamic changes of the immunoglobulins in patients with severe ovarian hyperstimulation syndrome: efficacy of a novel treatment using peritoneo-venous shunt. AB - PROBLEM: To evaluate the efficacy of continuous auto-transfusion system of ascites (CATSA) for the treatment of patients with severe ovarian hyperstimulation syndrome (OHSS) at the risk of febrile morbidity, the dynamic changes of immunoglobulins in the sera and the peritoneal fluid from patients with severe OHSS treated by CATSA were estimated. METHOD OF STUDY: Ten patients with severe OHSS after superovulation for in vitro fertilization-embryo transfer (IVF-ET) were treated by CATSA. Immunoglobulin concentrations were examined in the serum and in the peritoneal fluid before and after CATSA. As controls, serum samples from 15 infertile women, who did not develop OHSS after the same superovulation protocol, were obtained on the day of mid-luteal period (Control 1). Serum samples from 15 patients with OHSS, who were treated by albumin infusion without paracentesis, were also obtained before and after the treatment (Control-2). RESULTS: Before the treatments, serum immunoglobulin G (IgG) concentrations in patients with severe OHSS treated with CATSA and those in patients of Control-2 were significantly lower than those in patients of Control 1 (P < 0.01). Following CATSA, the concentration of IgG increased in the sera, while it decreased in the peritoneal fluid. CONCLUSIONS: Serum IgG in patients with severe OHSS exuded into their peritoneal cavity, indicating that they might be at the status of immunodeficiency and at the risk of febrile morbidity. However, non-infectious febrile morbidity attributed to endogenous pyrogenic mechanism might be considerable. It is also suggested that CATSA might be effective in improving hypoimmunoglobulinemia of the patients with severe OHSS by the peritoneo-venous shunt. PMID- 11883747 TI - Value of flow cytometric assay for the detection of antisperm antibodies in women with a history of recurrent abortion. AB - PROBLEM: To verify the proposed relationship between recurrent spontaneous abortions and the presence of maternal antisperm antibodies (ASA) in women as detected by a sensitive and reliable method. METHOD OF STUDY: The presence of maternal antipaternal immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies were determined against three different paternal antigens comprising T, B lymphocytes and semen cells by a sensitive flow cytometric crossmatch method to examine their possible correlation with pregnancy outcome. Group 1 consisted of sera obtained from 24 women with a history of abortion, and lymphocytes and semen samples collected from their husbands at the same time of visiting the in vitro fertilization (IVF) Clinic at King Faisal Specialist Hospital and Research Center. Sera, lymphocytes and semen samples were also collected from six couples with no history of abortion who served as controls (Group 2). RESULTS: Using a sensitive flow cytometric assay to analyse the samples, without knowledge of clinical status, elevated levels of both IgG and IgM were detected in Group 1. However, no significant association was found when compared with normal females who had healthy pregnancies. CONCLUSION: Flow cytometry is a highly sensitive and specific tool for the detection of alloantibodies in human sera from patients with rejected transplanted organs. Our findings suggest that maternal antipaternal antibodies with respect to IgG and IgM classes do not play a major role in women with a history of recurrent abortions, despite the presence of increased levels of antibodies against three different sources of paternal antigens. PMID- 11883748 TI - Testis-specific antigen (TSA-1) is expressed in murine sperm and its antibodies inhibit fertilization. AB - PROBLEM: We recently cloned and sequenced a sperm-specific antigen, designated as testis-specific antigen-1 (TSA-1), from human testis. The present study was conducted to examine its expression and function in murine sperm, in order to find out whether or not the mouse can provide a suitable model for examining its immunocontraceptive effects. METHOD OF STUDY: The antibodies (Ab) were raised against purified human rTSA-1 in virgin female rabbits. The rTSA-1 was run in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and the gel containing the approximately 18 kDa band was cut, minced and used for immunization to obtain the specific Ab. The immunoglobulins from preimmune bleed and from animals injected with adjuvant alone served as control. These Ab were analysed in enzyme-linked immunosorbent assay (ELISA), Western blot procedure, immunoprecipitation procedure, immunocytochemical technique (ICT), immunobead binding technique (IBT), acrosome reaction and sperm-zona binding assay. RESULTS: Active immunization of female rabbits with purified rTSA-1 protein of 18 kDa, produced high titer Ab against the recombinant antigen. These Ab to rTSA-1 were used in the present study. In Western blot procedure, rTSA-1 Ab recognized a specific protein band of approximately 24 +/- 3 kDa in murine sperm extract, the band similar to found in human sperm extract. In the immunoprecipitation procedure, rTSA-1 Ab immunoprecipitated the protein band of similar size from extracts of murine sperm and murine testis. The ICT and the IBT studies revealed the subcellular localization of TSA-1 on the surface of acrosome and tail regions of the non-capacitated and capacitated murine sperm cells. In functional bioassays, rTSA-1 Ab inhibited the acrosome reaction and sperm-egg binding in vitro. CONCLUSIONS: These data indicate that the TSA-1 is expressed in murine sperm and may have a biological role in sperm function and sperm-egg binding. In vitro inhibition of capacitation/acrosome reaction and sperm-zona binding suggests that the mouse can provide a suitable model to examine the immunocontraceptive effects of TSA-1 in actively immunized animals. PMID- 11883750 TI - Modulation of the uterine response to infectious bacteria in postpartum ewes. AB - PROBLEM: Exogenous progesterone and prostaglandin E2 (PGE2) can downregulate uterine immune functions and render the uterus susceptible to bacterial infection. METHOD OF STUDY: Ewes were sham-ovariectomized (SHAM) or ovariectomized (OVEX) 9 days after parturition (day 0), and their uteri were inoculated with Arcanobacterium pyogenes and Escherichia coli on day 15. Vena caval blood was collected on day 14 and days 16-19, and uteri were collected on day 20. Ewes began receiving either canola oil (OIL) or progesterone in oil (PROG) on day 10. Lymphocytes from each blood sample were assigned to a 2 x 2 factorial array of in vitro treatments; 10(-7) M PGE2 and 10(-7) M indomethacin (INDO) were main effects. [3H]Thymidine incorporation (expressed in picomoles) was used to quantify proliferation. RESULTS: Progesterone was greater (P = 0.001) in PROG than in OIL ewes (3.6 versus 0.7 ng/mL), and only PROG ewes developed infections. Lymphocyte proliferation was least (P = 0.02) in PROG-OVEX ewes (4.1 versus 5.4, 5.7, and 5.8 pmol for OIL-SHAM, PROG-SHAM, and OIL-OVEX, respectively). Concanavalin A (Con-A)-stimulated proliferation was less (P < 0.01) for PGE2- and PGE2 + INDO-treated lymphocytes (7.5 and 8.3 pmol, respectively) than for control or INDO-treated cells (12.9 and 14.7 pmol, respectively). CONCLUSIONS: Progesterone treatment of postpartum ewes suppressed uterine immunity. In vitro PGE, treatment suppressed lymphocyte proliferation, regardless of PROG, and highlights a progesterone-independent level of regulation of uterine immune function. PMID- 11883749 TI - Sperm immobilizing antibodies in the sera of infertile women cause low fertilization rates and poor embryo quality in vitro. AB - PROBLEM: The effects of sperm immobilizing antibodies in the sera of infertile women on fertilization and embryo quality in vitro were investigated. METHOD OF STUDY: Before the introduction of sperm immobilization test (SIT) as a routine test for female infertility, 85 oocytes were collected in nine in vitro fertilization (IVF) cycles from four infertile women who were afterward found having had sperm immobilizing antibodies in their sera and the oocytes were inseminated with swim-up sperm in a medium containing the patient's serum. Fifty oocytes were collected in five IVF cycles from five infertile women possessing the antibodies in their sera and the oocytes were inseminated with swim-up sperm in a medium supplemented with human serum albumin (HSA). RESULTS: In the former group, 41 of 85 oocytes were fertilized, giving a fertilization rate of 48.2%. In the latter group, 43 of 50 oocytes were fertilized, giving a fertilization rate of 86.0%. There was a significant difference of the fertilization rate between the groups (P < 0.0001). Embryo quality was assessed by the Veeck's classification. The grade 1 and grade 2 embryos were considered good quality. Using this classification, 16 (39.0%) of 41 embryos incubated in the medium containing the patient's serum were good quality, while 34 (79.1%) of 43 embryos incubated in the medium supplemented with HSA were good quality. There was also a significant difference between the groups (P = 0.0003). CONCLUSIONS: These findings might indicate that sperm immobilizing antibodies in the sera of infertile women cause low fertilization rates and poor embryo quality in vitro. It is suggested that SIT in the sera of infertile women should be performed at least before proceeding IVF. The manipulation of gametes and embryos from patients having sperm immobilizing antibodies should be carefully carried out especially to avoid contaminating patient's serum and follicular fluid in the culture medium in order to have a better IVF result. PMID- 11883751 TI - Blood lead reference values: the results of an Italian polycentric study. AB - This paper presents the results of a polycentric study carried out in seven different areas, organized by the Italian Society of Reference Values (SIVR) for assessing reference values of lead in blood (B-Pb) at the current doses of the metal to general population. The estimated arithmetic mean for B-Pb in males was of 45.1 microg/l and 30.6 microg/l in females; the 95th centile was 100 and 60 for males and females, respectively. The main variables influencing B-Pb levels were gender, age, BMI, outside sport practice, alcohol consumption and smoking habits, while the geographic area and the urban residence did not affect the metal concentration in blood. PMID- 11883752 TI - The importance of organic matter distribution and extract soil:solution ratio on the desorption of heavy metals from soils. AB - The lability (mobility and bioavailability) of metals varies significantly with soil properties for similar total soil metal concentrations. We studied desorption of Cu, Ni and Zn, from 15 diverse, unamended soils. These studies included evaluation of the effects of soil:solution extraction ratio and the roles of soil properties on metal desorption. Dcsorption was examined for each metal by computing distribution coefficients (Kd) for each metal in each soil where Kd = [M]soil/[M]solution, Results from soil:solution ratio studies demonstrated that Kd values for the metals tended to increase with increasing soil:solution ratio. This result also held true for distribution of soil organic matter (SOM). Because the soil:solution ratio has a significant effect on measured metal distributions, we selected a high soil:solution ratio to more closely approach natural soil conditions. Copper showed strong affinity to operationally defined dissolved organic matter (DOM). In this study, DOM was operationally defined based on the total organic carbon (TOC) content in 0.45 microm or 0.22-microm filtrates of the extracts. The Kd of Cu correlated linearly (r2 = 0.91) with the Kd of organic matter (Kd-om) where the Kd-om is equal to SOM as measured by Walkley-Black wet combustion and converted to total carbon (TC) by a factor of 0.59. These values representing solid phase TC were then divided by soluble organic carbon as measured by TOC analysis (DOM). The conversion factor of 0.59 was employed in order to construct Kd-om values based on solid phase carbon and solution phase carbon. SOM plays a significant role in the fate of Cu in soil systems. Soil-solution distribution of Ni and Zn, as well as the activity of free Cu2+, were closely related to SOM, but not to DOM. Kd values for Ni, Zn and free Cu2+ in a particular soil were divided by the SOM content in the same soil. This normalization of the Kd values for Ni, Zn, and free Cu2+ to the SOM content resulted in significant improvements in the linear relationships between non-normalized Kd values and soil pH. The semi-empirical normalized regression equations can be used to predict the solubility of Ni and Zn and the activity of free Cu2+ as a function of pH. PMID- 11883753 TI - Exposure to genotoxins present in ambient air in Bangkok, Thailand--particle associated polycyclic aromatic hydrocarbons and biomarkers. AB - Exposure to genotoxic compounds in ambient air has been studied in Bangkok, Thailand, by analysis of polycyclic aromatic hydrocarbons (PAHs) associated with particles and using different biomarkers of exposure. Eighty-nine male, non smoking Royal Thai police officers were investigated. The police officers were divided into a high exposure group (traffic police) and low exposure (office duty). Particulate matter was collected using personal pumps (2 l/min) and the eight carcinogenic PAHs were analysed by standard procedures. The traffic police was exposed to a 20-fold higher level of total PAHs than office police (74.25 ng/m3 vs. 3.11; P= 0.001). A two-fold variation was observed between the different police stations. The major PAHs in all groups was benzo[g,h,l]pyrelene. Large inter-individual differences in biomarker levels were observed, but the level of all markers was statistically significantly higher in the traffic police group than in the office group. The level of 1-hydroxypyrene (1-HOP) was 0.181+/ 0.078 (range 0.071-0.393) micromol/mol creatinine in the traffic group and 0.173+/-0.151 (P = 0.044) in the office group. The bulky carcinogen DNA-adduct level, determined by P32-post-labelling, was 1.6+/-0.9 (range 0.4-4.3) adducts/10(8) nucleotides in the traffic group and 1.2+/-1.0 (0.2-4.9) in the office group (P = 0.029; Mann-Whitney U-test). The serum PAH-albumin adduct level was 1.76 (0.51-3.07) fmol adducts/microg albumin in the traffic group and 1.35+/ 0.77 (0.11-3.45; P = 0.001) in the office group. Lower biomarker levels were observed during the period when the traffic police officers were wearing a simple facemask, indicating that these masks protect against particle-associated PAHs. No statistically significant correlations were observed between biomarker levels and the level of individual PAHs or total PAH. Our data show, that people in Bangkok, who spend most of the day outside air-conditioned offices, are exposed to high levels of genotoxic PAHs. However, for people who spend their working day in offices, the exposure is similar to people living in other metropolitan areas. PMID- 11883754 TI - Trace elements in foods and meals consumed by students attending the faculty cafeteria. AB - The content and intake of some trace elements (Pb, Cd, Ni, Hg, Cr) in meals, consumed by students attending the Faculty Cafeteria, were assessed. The study was carried out over 6 days of the 2nd week of February, 1993 and in three consecutive days of the second week of May. In those periods 10453 and 4055 students attended the cafeteria, respectively. After recording the ingredients and the preparation and cooking methods, some edible portions of foods and dishes were collected, homogenised, lyophilised and kept in polyethylene bottles until analysis. We found few differences in food content and dietary intake of some (Ni and Cr) but not all trace elements between February and May. Higher levels of Pb, Cd, Ni, Cr were present in bread (21.5+/-14.0, 5.6+/-0.02, 55.6+/-1.7 and 66.9+/ 0.1 microg/100 g of edible portion, respectively), followed by meat, filled pasta and cheese, whereas lower values were observed in vegetables and fruit. The highest Hg content was recorded in the pasta group. Among the most frequently consumed foods and meals, the highest levels of P, Pb, Cr, Hg, Ni were found in bread, meat and pasta. The calculation of mean trace element intake, corrected for leftovers, showed pasta, bread and meat as the main sources of Pb, Cd, Ni, Hg and Cr over the two periods. The estimated potential tolerated weekly intake (PTWI) resulted high for Pb and Cd and low for Hg. Finally, a large variability in trace element content between raw and cooked foods was observed. These results give valuable information on trace element content of foods to be used for preparation of meals at the Faculty cafeteria in different seasons. PMID- 11883755 TI - Concentration of atmospheric particulates during a dust storm period in central Taiwan, Taichung. AB - In this study we monitored concentrations of particles in central Taiwan using PS 1 (GPS1 PUF Sampler) and Model 310 Universal Air Sampler (UAS) from 02/23/2001 to 03/12/2001 at two sampling sites. During this period, an Asian dust storm moved across central Taiwan from 3/3 to 3/6. The total ambient air particle concentrations during the dust storm period were than compared with previous data from this region. In general, the average total suspended particulate (TSP) concentration order was during dust storm period > after dust storm period > non dust storm period at both HKITT (traffic) and THUC (rural) sampling sites. The ratio of PM2.5/PM10 was 60% before and after the dust storm period. However, this ratio was decreased to less than 50% during the dust storm. This demonstrates that the coarse particulate concentrations (PM2.5-10) increased during the dust storm period. In contrast the increase of ambient air particles concentrations after the Taiwan Chi-Chi Earthquake were mainly due to fine particles (PM2.5). And, the increased of ambient air particles concentrations after dust storm period were mainly coarse particle (PM2.5-10) concentrations in central Taiwan. PMID- 11883756 TI - Oral manganese intake estimated with dietary records and with direct chemical analysis. AB - The estimation of the daily oral intake of manganese (Mn) can vary significantly with the method used. This study aims to compare two different approaches: the use of dietary records along with tables of the Mn contents of different foods and the parallel collection of identical aliquots which are analyzed for Mn. Dietary samples and information were collected during a 3-day dietary assessment with five participants. The average daily oral intake of Mn estimated from the dietary records and tables of Mn contents was 3.52 mg/day (S.D. = 1.9), which corresponds to an exposure dose (ED) of 50.27 microg/kg body wt./day (S.D. = 26.2). With a parallel collection of identical aliquots and chemical analysis, the average intake was 2.88 mg/day (S.D. = 0.64) for an ED of 41.18 microg/kg body wt./day (S.D. = 8.9). Thus, the use of tables gives an estimated oral dose 22% higher than the more accurate direct chemical analysis. PMID- 11883757 TI - Correlation between mercury and selenium concentrations in Indian hair from Rondjnia State, Amazon region, Brazil. AB - Total mercury and selenium concentrations were determined in hair samples collected from Wari (Pacaas Novos) Indians living in Doutor Tanajura village, Gujara-Mirim city, Rondjnia State. The mercury concentrations in some samples are much higher than the values determined in samples from individuals not exposed to mercury contamination, occupationally or environmentally. The selenium concentrations are in the normal range. A correlation was observed between the mercury and selenium concentration and the values of the molar ratio approach 1 at low Hg concentrations. This fact is related to the equimolar complex formed by [(Hg-Se)n]m-Seleprotein P, which can decrease the bioavailable mercury in the organism. PMID- 11883758 TI - Factors affecting individual exposure to NO2 in Genoa (northern Italy). AB - The individual exposure to nitrogen dioxide (NO2) of 89 volunteers living in Genoa, a large port city of northern Italy, was investigated with personal passive diffusion tubes in February-March 2000. The data were related to NO2 concentration in the kitchen and bedroom as measured by static samplers. Volunteers included students, workers and housewives living in three areas of Genoa differing by street traffic and industrial plant location. The kitchen samples showed higher (47.00+/-16.5 microg/m3) NO2 concentrations than those from the bedroom (24.78+/-9.8 microg/m3); overall indoor NO2 concentrations were lower in the Eastern area of Genoa, where outdoor pollution is lower. Students were the volunteer group with the lowest exposure rate (24.9+/-7.8 microg/m3 vs. 44.3+/ 10.1 microg/m3 for workers and 40.0+/-13.4 microg/m3 for housewives). This difference is related to the fact that students spend more time indoors, where pollution levels are lower. The main household characteristics which were shown to affect personal NO2 exposure were (a) the presence of a chimney equipped with an active aspiration device in the kitchen and (b) the heating system. PMID- 11883759 TI - Metal contamination in sediments from a desalination plant effluent outfall area. AB - To assess the impact of seawater desalination effluent discharges on the receiving water body, the outfall area of a small desalination facility on the northwestern coast of the Arabian Gulf (Saudi Arabia) was investigated for metal contamination. Sediment samples were collected from a 6 x 6 km2 area and were analyzed for metal concentrations. Cadmium, cobalt, copper, mercury, vanadium, iron, phosphorus and zinc were very high in the sediment samples from the immediate vicinity of the outfall, and decreased progressively away from it. Contour maps of elemental concentrations confirm the above conclusions. Barium and chromium showed a decreasing trend towards the outfall, but this could be related to drilling activities in a nearby oil field (barium and chromium were high in the drilling mud). Concentrations of nickel, lead, and titanium exhibited no general trend in the sediment samples. PMID- 11883760 TI - Mercury speciation in the French seasonal snow cover. AB - Snow samples have been collected in the French Alps in 1998, 1999 and 2000 in order to measure both total Hg (HgT) and reactive Hg (HgR). Concentrations of HgT were between 13 and 130 pg g(-1) and HgR concentrations were below the detection limit (approximately 0.8 pg g(-1)). Hg speciation in snow was evaluated on the basis of ionic complexation equilibrium with chloride, hydroxide, oxalate. The pH of the snow was found to be an important parameter for Hg speciation. For pH values near 3, HgC2O4 is predominant in snow samples except for snow strongly influenced by anthropogenic sources (in which case HgCl2 predominates). When pH > 4, Hg(OH)2 and HgOHCl are predominant. These latter pH values are observed for precipitation not influenced by anthropogenic sources but more by soil erosion, e.g. Saharan dusts. The knowledge of Hgr speciation in snow is a key question for understanding the mechanisms of transformation of these complexes in snow after precipitation. PMID- 11883761 TI - Exposure of South Carolinians to commercial meats and fish within their meat and fish diet. AB - There has been considerable interest in the public's exposure to a variety of contaminants through the consumption of wild fish and game, yet there is little information on consumption of commercial meats and fish, or the relationship between commercial and self-caught fish. We conducted a dietary survey in 1999 to estimate exposure levels of 464 individuals from people attending the Palmetto Sportsmen's Classic. Mean consumption was similar for beef, chicken/turkey, and wild-caught fish, and much lower for pork and store-bought fish, and still lower for restaurant fish. There were no ethnic differences in the consumption of most commercial fish and meats, although the differences for chicken approached significance. There were significant ethnic differences in consumption of wild caught fish. Women ate significantly less of all meat types, except store-bought fish. People over 45 ate less beef than younger people, and people younger than 32 ate significantly more chicken than others. There were no significant differences in consumption patterns as a function of income, except for chicken and wild-caught fish; people with higher incomes ate more chicken than others, and people with lower incomes ate more wild-caught fish than others. When all wild-caught and commercial fish and meats are considered, there are significant differences only for ethnicity and gender. Blacks consume significantly more fish than Whites, and men consume significantly more than women. PMID- 11883763 TI - Large particle flux of 239+240Pu on the continental margin of the East China Sea. AB - Settling particles were collected from three locations in the East China Sea continental margin and analyzed for 239+240Pu. Two types of sediment traps were used, cylindrical traps and conical time-series traps. Surface sediment samples collected from five locations were also analyzed for 239+240Pu. Data from cylindrical traps showed there was a clear tendency for total mass fluxes to increase with depth at all three stations, and there was an especially large increase near the bottom. 239+240Pu concentrations in settling particles increased with depth from 1.76 mBq/g at 97-m depth to 3.0 mBq/g at 120-m depth and ranged from approximately 3 to 4 mBq/g at depths greater than 120 m. 239+240Pu concentrations collected in the near-bottom traps were approximately two times higher than those in the underlying surface sediments. Like total mass fluxes there was a clear tendency for 239+240Pu fluxes to increase with depth at every station, and the highest 239+240Pu fluxes were observed near the bottom. 239+240Pu concentrations in the time-series traps had little variation throughout the sampling period, though the total mass fluxes showed a large variation. A high variability of 239+240Pu fluxes occurred in very short period of time (1/2 day). The large fluxes of 239+240Pu might be attributed to episodic lateral transport of particles that flow down the continental slope with the nepheloid layer which was considered to be significant for 239+240Pu transport on the continental slope in the East China Sea. PMID- 11883762 TI - Catchment-specific element signatures in estuarine crocodiles (Crocodylus porosus) from the Alligator Rivers Region, northern Australia. AB - The concentrations of Na, K, Ca, Mg, Ba, Sr, Fe, Al, Mn, Zn, Pb, Cu, Ni, Cr, Co, Se, U and Ti were determined in the osteoderms and/or flesh of estuarine crocodiles (Crocodylus porosus) captured in three adjacent catchments within the Alligator Rivers Region (ARR) of northern Australia. Results from multivariate analysis of variance showed that when all metals were considered simultaneously, catchment effects were significant (P < or = 0.05). Despite considerable within catchment variability, linear discriminant analysis (LDA) showed that differences in elemental signatures in the osteoderms and/or flesh of C. porosus amongst the catchments were sufficient to classify individuals accurately to their catchment of occurrence. Using cross-validation, the accuracy of classifying a crocodile to its catchment of occurrence was 76% for osteoderms and 60% for flesh. These data suggest that osteoderms provide better predictive accuracy than flesh for discriminating crocodiles amongst catchments. There was no advantage in combining the osteoderm and flesh results to increase the accuracy of classification (i.e. 67%). Based on the discriminant function coefficients for the osteoderm data, Ca, Co, Mg and U were the most important elements for discriminating amongst the three catchments. For flesh data, Ca, K, Mg, Na, Ni and Pb were the most important metals for discriminating amongst the catchments. Reasons for differences in the elemental signatures of crocodiles between catchments are generally not interpretable, due to limited data on surface water and sediment chemistry of PMID- 11883764 TI - Threshold, comfortable level and impedance changes as a function of electrode modiolar distance. AB - OBJECTIVE: The study investigated the hypothesis that threshold and comfortable levels recorded from cochlear implant patients would reduce, and dynamic range increase, as distance of the electrode from the modiolar wall (radial distance) decreases. Two groups of cochlear implant patients participated; one group using the Nucleus' 24 Contour electrode array, and one group using the Nucleus standard straight (banded) array. The Nucleus 24 Contour array has been shown in temporal bone studies to lie closer to the modiolus than the banded array. The relationship of electrode impedance and radial distance is also investigated. DESIGN: The study, conducted at three centers, evaluated 21 patients using the Contour array, and 36 patients using the banded array. For each patient, threshold, comfortable levels and dynamic range were measured at four time points. Common ground electrode impedance was recorded clinically from each patient, at time intervals up to 12 wk. An estimate of the radial distance of the electrode from the modiolus was made by analysis of Cochlear view x-rays. RESULTS: Threshold and comfortable levels were significantly lower for the Nucleus 24 Contour array than for the banded array. However, dynamic range measurements did not show the predicted increase. In a majority of subjects, a significant correlation was found between the estimated radial distance of the electrode from the modiolus and the measured threshold and comfortable levels. This trend was not observed for dynamic range. The analysis indicates that other factors than radial distance are involved in the resultant psychophysical levels. Clinical impedance measures (common ground) were found to be significantly higher for the Contour array. However, the electrodes on the Contour array are half rings, which are approximately only half the geometric size of the full rings as electrodes of the standard array. When the geometric electrode area in the two array designs are normalized, the trends in the electrode impedance behavior are similar. CONCLUSIONS: The results support the hypothesis that the relationship between the radial distance of the electrode and the psychophysical measures are influenced by patterns of fibrous tissue growth and individual patient differences, such as etiology and neural survival. Impedance measures for the Nucleus 24 Contour electrode array were higher than the banded electrode array, but this is primarily due to the reduction in electrode surface area. The different outcomes in impedance over time suggest differences in the relative contributions of the components of impedance with the two arrays. PMID- 11883765 TI - Nucleus 24 advanced encoder conversion study: performance versus preference. AB - OBJECTIVE: The Nucleus 24 Advanced Encoder Conversion Study was designed to determine the safety and effectiveness of the advanced combination encoder (ACE) and continuous interleaved sampling (CIS) speech coding strategies compared with that of the spectral peak (SPEAK) strategy in a large sample of postlinguistically deaf adults. Data from this study were analyzed to test the hypothesis that the group of subjects who prefer a given strategy for use in everyday life obtain significantly higher speech recognition scores (as a group) with the preferred strategy than with nonpreferred ones. DESIGN: The first 100 adults implanted with the Nucleus 24 Cochlear Implant System who had a minimum of 3 mo experience with the device were invited to participate. Those who accepted were randomly assigned to one of two groups for an initial 6-wk use of either the ACE or the CIS strategy; the other strategy was used during the second 6-wk period. Parameters in subjects' SPrint speech processor programs were adjusted to maximize perceived benefit with each strategy in everyday life. Recognition of medial consonants and vowels, CNC words, CUNY sentences in quiet and at + 10 dB signal to noise ratio, and HINT sentences in quiet was initially evaluated at the beginning of the study with the SPEAK strategy and at the end of the two 6-wk periods with the ACE and CIS strategies. Then subjects' processors were programmed with all three strategies for use in everyday life. After 3-wk use, a final evaluation of speech recognition with the HINT sentences in quiet and CUNY sentences at +10 dB signal to noise ratio was performed with each strategy. Subjects also responded to a questionnaire giving their strategy preference for most listening situations, the percentage of time they used each strategy, and the strategy they found gave them the best hearing and understanding of speech in 19 listening situations. RESULTS: Of the 62 subjects who participated, 56 subjects reported that they preferred one strategy for most listening situations (ACE strategy: 37 [59.7%]; SPEAK strategy: 14 [22.6%]; CIS strategy: 5 [8.0%]) and six subjects did not prefer a single strategy (9.7%). For the group who preferred one strategy, the preferred strategy resulted in higher scores than for one of the other strategies at the initial evaluation on CUNY sentences in quiet and noise and at the final evaluation on HINT sentences in quiet and CUNY sentences in noise for approximately two-thirds of the subjects. Strategy preference and performance were not significantly related for the remaining dependent measures. There also was strong agreement between the preferred strategy, percentage time this strategy was used, and the number of specific listening situations the preferred strategy was chosen for best hearing and understanding of speech. Although the majority of subjects strongly preferred a single strategy, some preferred to use two or three strategies, and a few were not sure which strategy they preferred for the majority of listening situations. Of the 19 subjects who reported that it was useful to use different strategies for different listening situations, only 5 of the 13 subjects, who responded to a follow-up questionnaire sent 18 mo later, continued to use multiple strategies. CONCLUSIONS: There was a significant relation between subjects' strategy preference based on experience in everyday life and their performance on the sentence tests, particularly sentences in noise. Important individual differences in strategy preference as well as in rate and number of channels stimulated per cycle within the ACE and CIS strategies emerged during the study. At the end of this process, over half of the subjects preferred the ACE strategy, and over double the number preferred the SPEAK strategy compared with the CIS strategy. To provide newly implanted recipients with as much benefit as possible, it is important that the speech processor program with each strategy be adjusted to maximize perceived benefit sequentially and then the three strategies need to be compared. With the four memories of the SPrint processor and a recipient who adapts quickly to hearing sound with different speech coding strategies, it may be possible to accomplish this comparison clinically through weekly fitting sessions plus listening in everyday life over a period of approximately 6 wk. At the end of this fitting process, most recipients probably will prefer to use one strategy, whereas some may prefer two or all three strategies to maximize their ability to hear in different listening situations. PMID- 11883766 TI - The nucleus 24 contour cochlear implant system: adult clinical trial results. AB - OBJECTIVE: The purpose of this article is to present psychophysical data for 40 Nucleus 24 Contour adult patients with 1 mo of device experience and speech perception results for a group of 56 adult patients with 3 mo experience using the Nucleus 24 Contour cochlear implant system. Postoperative hearing thresholds (i.e., under headphones) in the implanted ear were also assessed in a group of 85 patients who had measurable hearing preoperatively. This was of interest because preservation of residual hearing, postoperatively, is consistent with atraumatic insertion of the electrode array. In addition, data will be presented that reflected feedback from 40 surgeons who participated in the trial. DESIGN: Participants in this study were 18 yr of age or older, with bilateral severe to profound sensorineural hearing loss with no congenital component. Preoperatively, they scored < or = 50% open-set sentence recognition (HINT sentences) in the ear to be implanted and < or = 60% in the best-aided condition. The investigation was a repeated-measures single-subject experiment and took place at 46 different North American clinical sites. Preoperative performance was compared with postoperative performance 3 mo after device activation. Clinicians were able to program patients' processors with one, two, or all three speech-processing strategies. Testing took place using the participant's preferred speech processing strategy (SPEAK, CIS, or ACE). Preoperative unaided hearing thresholds were compared with unaided thresholds in the implanted ear measured 1 mo after device activation. Surgeons were canvassed regarding surgical use and design of the device via a questionnaire after having completed at least one Nucleus 24 Contour surgery. RESULTS: Average T- and C-levels for the Nucleus 24 Contour patients were considerably lower than those using the Nucleus 24 (CI24M). A total of 85 patients had measurable hearing preoperatively at two or more audiometric frequencies in the ear implanted. Of these patients 41 (48%) had measurable hearing at one or more frequencies and 32 (38%) had measurable hearing at two or more frequencies postoperatively. In general, surgeons found the Nucleus 24 Contour easy to insert and were pleased with the design features of the device. The downsized receiver/stimulator (of the Nucleus 24 Contour) required less drilling than the Nucleus 24, reducing surgical time, as well as making the Contour better suited for implantation in those with small skull sizes (e.g., small children and infants). After 3 mo of device use, mean open-set speech perception in quiet and in noise was significantly better than preoperative performance on all test measures. Patients using the ACE strategy had significantly better mean scores for all measures than patients using SPEAK. Only two patients preferred to use the CIS coding strategy. CONCLUSIONS The results presented in this article demonstrated that the design objectives of the Nucleus 24 Contour were met. Namely, results from this study, together with insertion studies, were consistent with perimodiolar placement using an implant design that the majority of surgeons found easy to insert with relatively minimal trauma. Reduced T- and C-levels were observed with Contour patients when compared with patients using the Nucleus 24 with the straight array, consistent with perimodiolar placement. A survey of surgeons participating in the clinical trial indicated easier, or equally easy, insertion of the Contour array, compared with previous Nucleus products as well as other manufacturers' devices, without the use of additional insertion tools or array positioners. Postoperatively, 46% of patients with preoperative residual hearing maintained some level of unaided hearing postoperatively, suggesting atraumatic insertion of the Nucleus 24 Contour electrode array. It is worth noting that all 216 patients implanted during this study had full insertions of their Contour electrode arrays. High levels of open-set speech perception in quiet and in noise were achieved and patients using the ACE strategy had significantly better mean scores for all measures than patients using SPEAK. Only two patients preferred to use the CIS coding strategy. PMID- 11883767 TI - Adaptive dynamic range optimization for cochlear implants: a preliminary study. AB - OBJECTIVE: The present study investigated the acceptability and the effect of Adaptive dynamic range optimization (ADRO) on speech perception for cochlear implant subjects. ADRO is a preprocessing scheme that continuously adjusts the gain in each frequency band to optimize the signal in the output dynamic range. DESIGN: Speech processor programs were created with and without ADRO processing. Nine subjects were tested in the laboratory and encouraged to use both programs in everyday listening situations. Take-home experience was assessed with preference questionnaires. Speech perception performance was compared for the standard and ADRO programs using City University of New York (CUNY) sentences, consonant-nucleus-consonant (CNC) words, and closed set spondees presented in quiet. A range of presentation levels were used; from 70 to 40 dB sound pressure level (unweighted RMS). CUNY sentences were also presented in multi-talker babble with +15 dB and +10 dB signal to noise ratios. RESULTS: There was a significant improvement in speech perception scores with the ADRO programs compared with the standard. At 50 dB, the mean open set sentence scores in quiet improved by 16% (p < 0.001). There was an improvement in mean word score for CNC words presented at 60 dB of 9.5% (p < 0.001) and a 20% improvement in mean score for spondees presented at 40 dB. There was no significant difference in sentence scores between the ADRO and the standard program for sentences presented in either noise condition. ADRO was the preferred program in 59% of listening situations, with five out of nine subjects indicating a strong overall preference and three subjects indicating a slight preference for ADRO. CONCLUSIONS: Continual adjustment of channel gains using ADRO provided improved sound quality and improved speech perception performance. Therefore, ADRO is a viable alternative to fixed channel gain and offers a means for cochlear implantees to gain more benefit from their devices. PMID- 11883768 TI - Surgical technique for the Nucleus Contour cochlear implant. AB - This paper deals with the Nucleus C124R (CS) (Contour) cochlear Implant: its characteristics, differences compared with the previous generation of devices, the perimodiolar electrode, and the surgical technique used for safe insertion. We also discuss the rationale behind perimodiolar electrodes in general, as well as the results of laboratory studies validating the design and safety of this particular electrode array. The differences in surgical technique between this device and prior Nucleus cochlear implants are as follows: the incision and the size of the well, or recess, for the electronics are smaller; the cochleostomy is larger; the posterior portion is placed in a subpericranial pocket, not tied down, before electrode insertion; and the insertion process itself is quite different, due to the nature of the electrode, its size, shape, and stylet. The technique described is that used by one experienced cochlear implant center, and reflect the authors' practice. Clearly, there are other possible variations on this theme, which may be equally satisfactory in other hands. Most surgeons find this device to be easier to place than previous generations: complications to date have been uncommon. PMID- 11883769 TI - A model of a nucleus 24 cochlear implant fitting protocol based on the electrically evoked whole nerve action potential. AB - OBJECTIVE: The goal of this study was to model a cochlear implant fitting protocol based on the electrically evoked whole nerve action potential (EAP) measured using the Neural Response Telemetry capabilities of the Nucleus C124M cochlear implant. The model and data were based on the significant correlations found between the EAP threshold and growth function slope and psychophysical threshold and, dynamic range, respectively, in 12 subjects (Franck, Reference Note 3; Franck & Norton, 2001). DESIGN: Using a retrospective split-half study design, these correlations found between psychophysical mapping levels and EAP data from six of the subjects were used in a model to predict psychophysical mapping levels from EAP data of the other six subjects. RESULTS: Predicted psychophysical mapping levels from the model of the EAP-based fitting protocol closely approximated measured cochlear implant fitting psychophysics. CONCLUSIONS: The close approximation of the measured data to the model data indicates the feasibility of the clinical use of an EAP-based cochlear implant fitting protocol. The realization of this model would only require two loudness judgments from the patient, whereas traditional fitting requires 44, and would be fit in a live-voice mode, accounting for across-electrode loudness summation. PMID- 11883770 TI - Speech perception using maps based on neural response telemetry measures. AB - OBJECTIVE: To determine how speech perception is affected for Nucleus cochlear implant users when their speech processor is programmed using neural response telemetry (NRT) measures rather than traditional behavioral estimates of threshold and maximum comfort level. DESIGN: Electrically evoked compound action potential (EAP) thresholds were measured for a group of 10 adult Nucleus cochlear implant users. These physiologic threshold estimates were used to create three SPEAK MAPs. One MAP ("+10/-20 MAP") was created using only the NRT data. The second MAP ("Combined MAP") was created using a combination of both EAP thresholds and a single behavioralmeasure of threshold and maximum comfort level from electrode 10. A third MAP ("Measured MAP") was created using standard programming techniques. Speech perception was then assessed using each of these three MAPs for either CUNY or HINT sentences presented at two different presentation levels (70 dB SPL and 55 dB SPL). RESULTS: On average, at the higher presentation level (70 dB SPL) subjects performed significantly better when using the Measured MAP than whenusing either of the NRT-based MAPs. No significant difference in speech perception was obtained for either of the three MAPs when the lower presentation level (55 dB SPL) was used. CONCLUSION: This study demonstrated that at relatively high presentation levels, speech perception scores obtained by subjects who use MAPs based solely or primarily on EAP thresholds ar eslightly lower than similar scores obtained using a more traditional MAP. This difference did not reach statistical significance at lower presentation levels. We interpret these findings in a positive light to suggest that although NRT-based MAPs may not be optimal, they are of sufficient quality to support reasonable levels of speech understanding. This is important to establish because these MAPs may prove to be most useful when very young children receive a cochlear implant. PMID- 11883771 TI - Three-month results with bilateral cochlear implants. AB - OBJECTIVES: To evaluate possible binaural listening advantages for speech in quiet, speech in noise, and for localization in a group of postlingually deafened adults with two cochlear implants functioning independently after 3 mo experience. DESIGN: Nine postlingually deafened subjects who had received a Cochlear Corporation CI24M implant in each ear were evaluated on a number of tasks. The subjects all had audiometric or biographical (e.g., duration of deafness) differences between the ears. Word and sentence materials were presented to the subjects in quiet and in noise with the signal always in the front and the noise from the front or either side. Results are reported for each ear and for both ears with the noise on either side. This allowed evaluation of head shadow and squelch effects. Additionally, localization ability was assessed for broadband noise presented either to the right or left of center at 45 degrees azimuth. Localization was assessed for each ear and for both ears. RESULTS: Results of speech testing in quiet showed a significant advantage for the binaural condition over the better ear in four subjects. In noise, with both signal and noise in front of the subject, a significant advantage of two ears over the better ear was found for four subjects. For noise to one side of the head, when the ear opposite the noise source was added to the ear ipsilateral to the noise, a significant advantage was demonstrated for seven of seven tested subjects. When the ear ipsilateral to the noise was added to the ear contralateral to the noise, a significant advantage was shown for only one of seven (noise on right) and three of seven (noise on left) tested subjects. The localization task showed that all seven tested subjects could discriminate 45 degrees left from 45 degrees right above chance with bilateral stimulation. Three subjects could perform the discrimination above chance with only one ear. However, performance with both ears was significantly better than performance with one ear for two of these latter subjects. CONCLUSIONS: We conclude that bilateral cochlear implants can provide real advantages, particularly when it is possible to utilize the ear that is away from a noise source, thus taking advantage of the head shadow effect. In addition, localization ability was generally better with two implants than with one. PMID- 11883772 TI - The results in patients implanted with the nucleus double array cochlear implant: pitch discrimination and auditory performance. AB - OBJECTIVE: In patients with total or surgically inaccessible cochlear obliteration, only a reduced number of active electrodes can be inserted with standard cochlear implants, resulting in below average auditory performance. Therefore, a special implant with two electrode arrays was developed on the basis of the Nucleus 22 cochlear implant, the socalled Double Array. One electrode array with 11 active electrodes is inserted into the basal turn of the cochlea, while the second array with 10 active electrodes is inserted into the second turn. The Double Array is now available on the basis of the more advanced Nucleus 24 with 11 active electrodes on each array and two reference electrodes, one at the case and the second one an additional ball electrode, which is placed under the temporalis muscle. For device description and surgical technique see Lenarz et al. (2001). This paper presents psychophysical data on pitch discrimination and auditory performance of patients implanted with a Double Array on the basis of the Nucleus 22. STUDY DESIGN: A prospective intra-individual study using a Latin square paradigm was performed in six adult patients with obliterated cochlea who received the Nucleus 22 Double Array. After appropriate fitting and loudness balancing, patients were tested either with the basal, the apical or both electrode arrays. Apart from auditory performance tests including numbers and monosyllable word tests, pitch discrimination was determined with a defined procedure. RESULTS: When activating each array alone, auditory performance was better with the basal array than with the apical array. Both arrays together showed marked improvement compared with the basal array, indicating an additional effect of the second array. Pitch discrimination was significantly better for the electrodes in the basal turn than in the second turn, indicating differences in electrical excitation of the auditory nerve fibers. Pitch discrimination was positively correlated with auditory performance data. CONCLUSION: The additional apical array leads to significant improvement in auditory performance in patients with obliterated cochleae by increasing the number of intracochlear electrodes. Despite reduced pitch discrimination, the apical array provides important information for speech recognition. For this reason the Double Array provides a profound advantage for patients with obliterated or surgically inaccessible cochleae. PMID- 11883773 TI - A quick solution structure determination of the fully oxidized double mutant K9 10A cytochrome c7 from Desulfuromonas acetoxidans and mechanistic implications. AB - Lysines 9 and 10 in Desulfuromonas acetoxidans cytochrome c7, which could be involved in the interaction mechanism with the redox partners, have been replaced by alanine residues using site-directed mutagenesis. The solution structure of the fully oxidized form of K9-10A cytochrome c7, which is paramagnetic with three paramagnetic centers, has been determined via 1H NMR. The assignment of the spectra has been performed through an automatic program whose algorithm and strategy are here described. The assignment of the NOESY spectra has been further extended by back calculating the NOESY maps. The final number of meaningful NOE based upper distance limits was 1186. In the Restrained Energy Minimization calculations, 147 pseudocontact shift constraints were also included, which showed consistency with NOE-based constraints and therefore further contribute to validate the structure quality. A final family of 35 conformers was calculated with RMSD values with respect to the mean structure of 0.69 +/- 0.17 A and 1.05 +/- 0.14 A for the backbone and heavy atoms, respectively. The overall fold of the molecule is maintained with respect to the native protein. The loop present between heme III and heme IV results to be highly disordered also in the present structure although its overall shape mainly resembles that of the oxidized native protein, and the two strands which give rise to the short beta-sheet present at the N-terminus and connected by a turn containing the mutated residues, are less clearly defined. If this loop is neglected, the RMSD values are 0.52 +/- 0.07 A and 0.92 +/- 0.06 A for the backbone and heavy atoms, respectively, which represent a reasonable resolution. The relative distances and orientations of the three hemes are maintained, as well as the orientation of the imidazole rings of the axial histidine ligands, with the only exception of heme IV. Such difference probably reflects minor conformational changes due to the substitution of the vicinal Lys 10 with an Ala. The replacement of the two lysines does not affect the reduction potentials of the three hemes, consistently with the expectations on the basis of the structure and electrostatic calculations. However, the replacement of the two lysines affects the reactivity of the mutant cytochrome c7 with [Fe] hydrogenase, inducing a change in Km. This finding is in agreement with the identification of the protein area around heme IV as the interacting site. PMID- 11883774 TI - Efficiency of paramagnetism-based constraints to determine the spatial arrangement of alpha-helical secondary structure elements. AB - A computational approach has been developed to assess the power of paramagnetism based backbone constraints with respect to the determination of the tertiary structure, once the secondary structure elements are known. This is part of the general assessment of paramagnetism-based constraints which are known to be relevant when used in conjunction with all classical constraints. The paramagnetism-based constraints here investigated are the pseudocontact shifts, the residual dipolar couplings due to self-orientation of the metalloprotein in high magnetic fields, and the cross correlation between dipolar relaxation and Curie relaxation. The relative constraints are generated by back-calculation from a known structure. The elements of secondary structure are supposed to be obtained from chemical shift index. The problem of the reciprocal orientation of the helices is addressed. It is shown that the correct fold can be obtained depending on the length of the alpha-helical stretches with respect to the length of the non helical segments connecting the alpha-helices. For example, the correct fold is straightforwardly obtained for the four-helix bundle protein cytochrome b562, while the double EF-hand motif of calbindin D9k is hardly obtained without ambiguity. In cases like calbindin D9k, the availability of datasets from different metal ions is helpful, whereas less important is the location of the metal ion with respect to the secondary structure elements. PMID- 11883775 TI - Exact solutions for chemical bond orientations from residual dipolar couplings. AB - New methods for determining chemical structures from residual dipolar couplings are presented. The fundamental dipolar coupling equation is converted to an elliptical equation in the principal alignment frame. This elliptical equation is then combined with other angular or dipolar coupling constraints to form simple polynomial equations that define discrete solutions for the unit vector(s). The methods are illustrated with residual dipolar coupling data on ubiquitin taken in a single anisotropic medium. The protein backbone is divided into its rigid groups (namely, its peptide planes and Calpha frames), which may be solved for independently. A simple procedure for recombining these independent solutions results in backbone dihedral angles phi and psi that resemble those of the known native structure. Subsequent refinement of these phi-psi angles by the ROSETTA program produces a structure of ubiquitin that agrees with the known native structure to 1.1 A Calpha rmsd. PMID- 11883776 TI - Sequence-specific assignment of histidine and tryptophan ring 1H, 13C and 15N resonances in 13C/15N- and 2H/13C/15N-labelled proteins. AB - Methods are described to correlate aromatic 1H(delta)2/13C(delta)2 or 1H(epsilon)1/15N(epsilon)1 with aliphatic 13C(beta) chemical shifts of histidine and tryptophan residues, respectively. The pulse sequences exclusively rely on magnetization transfers via one-bond scalar couplings and employ [15N, 1H]- and/or [13C, 1H]-TROSY schemes to enhance sensitivity. In the case of histidine imidazole rings exhibiting slow HN-exchange with the solvent, connectivities of these proton resonances with beta-carbons can be established as well. In addition, their correlations to ring carbons can be detected in a simple [15N, 1H]-TROSY-H(N)Car experiment, revealing the tautomeric state of the neutral ring system. The novel methods are demonstrated with the 23-kDa protein xylanase and the 35-kDa protein diisopropyl-fluorophosphatase, providing nearly complete sequence-specific resonance assignments of their histidine delta-CH and tryptophan epsilon-NH groups. PMID- 11883777 TI - NMR-based structural characterization of large protein-ligand interactions. AB - Genomic research on target identification and validation has created a great need for methods that rapidly provide detailed structural information on protein ligand interactions. We developed a suite of NMR experiments as rapid and efficient tools to provide descriptive structural information on protein-ligand complexes. The methods work with large proteins and in particular cases also without the need for a complete three-dimensional structure. We will show applications with two tetrameric enzymes of 120 and 170 kDa. PMID- 11883778 TI - Measurement of conformational constraints in an elastin-mimetic protein by residue-pair selected solid-state NMR. AB - We introduce a solid-state NMR technique for selective detection of a residue pair in multiply labeled proteins to obtain site-specific structural constraints. The method exploits the frequency-offset dependence of cross polarization to achieve 13COi-->15Ni-->13Calphai transfer between two residues. A 13C, 15N labeled elastin mimetic protein (VPGVG)n, is used to demonstrate the method. The technique selected the Gly3 Calpha signal while suppressing the Gly5 Calpha signal, and allowed the measurement of the Gly3 Calpha chemical shift anisotropy to derive information on the protein conformation. This residue-pair selection technique should simplify the study of protein structure at specific residues. PMID- 11883779 TI - Sequence-specific chemical shift assignment and chemical shift indexing of murine apo-Mts1. PMID- 11883780 TI - Assignments of 1H, 13C, and 15N resonances of human lysozyme at 4 degrees C. PMID- 11883781 TI - Sequence-specific resonance assignment of the second Ran-binding domain of human RanBP2. PMID- 11883782 TI - Virtually complete 1H, 13C and 15N resonance assignments of the second family 4 xylan binding module of Rhodothermus marinus xylanase 10A. PMID- 11883783 TI - 1H, 13C, and 15N resonance assignment of the vascular endothelial growth factor receptor-binding domain in complex with a receptor-blocking peptide. PMID- 11883784 TI - 1H, 15N and 13C resonance assignments of rabbit apo-S100A11. AB - S100 proteins belong to the EF-hand family of calcium binding proteins. Upon calcium binding, these proteins undergo a conformational change to expose a hydrophobic region necessary for target protein interaction. One member of the S100 protein family is S100A11, first isolated from chicken gizzard and termed calgizzarin. It was later isolated from other organisms and tissues including human placenta, pig heart and rabbit lung. The physiological target of S100A11 is thought to be annexin I, a phospholipid-binding protein involved in EGF receptor sorting. This work reports the 1H, 15N and 13C resonance assignments of rabbit apo-S100A11 determined using 15N, 13C-labelled protein and multidimensional NMR spectroscopy. PMID- 11883785 TI - Backbone NMR assignments of ribosome recycling factors (RRFs) from Escherichia coli and Tthermotoga maritima. PMID- 11883786 TI - 1H, 13C and 15N resonance assignments for the perdeuterated 22 kD palm-thumb domain of DNA polymerase beta. PMID- 11883787 TI - Sequence specific resonance assignment of the central domain of cardiac myosin binding protein C (MyBP-C). PMID- 11883788 TI - Differences between SHR and WKY following the airpuff startle stimulus in the number of Fos expressing, RVLM projecting neurons. AB - The neurocircuitry responsible for excessive stress-induced cardiovascular responses in genetic hypertensive rats remains elusive. Prior studies detailed a differential cardiovascular response profile to airpuff startle stimuli between Spontaneously Hypertensive (SHR) and Wistar Kyoto (WKY) rats. We recently identified strain differential Fos expression in the rostroventrolateral medulla (RVLM) and several RVLM projecting sites following airpuff startle. The current study sought to define RVLM projecting neurons that also express Fos following placement in the test chamber and administration of the airpuff startle stimulus. Unilateral iontophoretic micro-injections of fluorogold were made into the RVLM of 9-10 week old SHR and WKY rats. Two to three weeks later, animals were subjected to a series of 60 airpuff startle stimuli. Brains were double labeled for Fos and fluorogold. Single fluorogold and single Fos cells, and double labeled cells were found in the nucleus tractus solitarius (NTS), caudal ventral lateral medulla (CVLM), Kolliker fuse (KF), ventral lateral, lateral, and dorsal central gray, lateral hypothalamus (LH), and paraventricular nucleus of the hypothalamus (PVN). These data are consistent with the notion that the RVLM receives differential excitatory and/or inhibitory input from higher brain centers, perhaps contributing to differential Fos expression in the RVLM, differential autonomic responding, or both. PMID- 11883789 TI - Testosterone increased blood pressure and decreased renal tyrosine hydroxylase activity in SHR/y and Wistar-Kyoto rats. AB - The present study evaluated the association between a testosterone-induced elevation in blood pressure (BP) and renal tyrosine hydroxylase activity in SHR/y and Wistar-Kyoto male rats. The SHR/y rat is a consomic strain having the Y chromosome of the Spontaneously Hypertensive Rat and autosomes and the X chromosome from normotensive Wistar-Kyoto (WKY). Rats were castrated at 4-6 weeks and divided into control and sham groups (n = 6/group) with testosterone and blank sham implants respectively. BP and blood were taken every 2 weeks for estimation of serum testosterone and catecholamines. The animals were terminated at 16-18 weeks and kidneys were removed for the estimation of tyrosine hydroxylase activity. The testosterone treated rats had higher BP, plasma testosterone levels, kidney weights, but lower renal tyrosine hydroxylase activity than the sham treated controls. Hence, chronic testosterone treatment inhibits renal tyrosine hydroxylase activity in WKY and SHR/y rats. PMID- 11883790 TI - Paradoxical decrease in plasma NOx by L-arginine load in diabetic and non diabetic subjects. AB - L-arginine, a substrate of nitric oxide synthase, was infused (30 g/300 ml/30 min) to patients with or without type 2 diabetes to examine whether or not endothelial dysfunction expressed as attenuated depressor response to the substrate in diabetic patients may accompany attenuated plasma NOx (NO2- and NO3 ; an index of NO formation) elevation. Decrease in blood pressure by L-arginine was significantly smaller in diabetic patients than that in non-diabetic patients, and increase in plasma cGMP level in diabetic patients tended to be smaller and retarded than non-diabetic patients. However, plasma NOx decreased in both groups in a similar degree without changes in urinary NOx excretion, implying that NOx in plasma moved to other compartments. These results indicate that plasma NOx could not be solely used as an index of NO formation by L arginine load and that this paradoxical decrease in plasma NOx would require further examination extending to other NOx compartments. PMID- 11883791 TI - Effects of losartan and benazepril on abnormal circadian blood pressure rhythm and target organ damage in SHRSP. AB - The effects of chronic treatment with losartan, an angiotensin II type 1 (AT1) receptor antagonist, and benazepril, an angiotensin converting enzyme (ACE) inhibitor, on target-organ damage and abnormal circadian blood pressure (BP) rhythm were compared in stroke-prone spontaneously hypertensive rats (SHRSP). Losartan and benazepril were given by intraperitoneal infusion for 3 weeks after 17 weeks of age to minimize any influence of their different pharmacokinetic properties. BP was continuously monitored by telemetrical method before treatment and at the end of the observation period. The left ventricular (LV) weight, 24 hour urinary albumin excretion (UalbV) and morphological changes in the kidney were observed. Losartan and benazepril (1, 3 and 10 mg/day) reduced BP and LV weight in a dose-dependent manner with good correlation between the effects. Losartan significantly improved UalbV in a dose-dependent manner, whereas benazepril was effective at only 10 mg/day. Renal morphological analysis showed that reduction of glomerulosclerosis and collagen fiber thickness was related to the effect on UalbV, but not to the antihypertensive effects. Losartan improved the shifted circadian BP rhythm towards the active phase in a dose-dependent manner, whereas the improvement caused by 1 and 3 mg/day of benazepril was less effective than the same dosage of losartan. These results suggest that both losartan and benazepril can reduce cardiac hypertrophy showing good correlation with their antihypertensive effects, but losartan, especially at a low dose, alleviates renal damage more effectively than benazepril, with its effect correlating well with improvement of the abnormal circadian BP rhythm in SHRSP. Thus, the protective effect against hypertensive target organ damage of the AT1 receptor antagonist seems to be more effective than that of ACE inhibitor. PMID- 11883792 TI - Beneficial effects of fenoldopam treatment on renal function in streptozotocin induced diabetic rats. AB - We have studied the effect of chronic treatment with dopamine D1 receptor agonist fenoldopam (1 mg/kg, i.p. daily for 6 weeks) on renal function and metabolic parameters in streptozotocin (STZ)-diabetic rats. Diabetes was induced by a single tail vein injection of STZ (45 mg/kg). STZ produced severe hyperglycemia, hypoinsulinemia, hypercholesterolemia, hypertriglyceridemia, hypertension and bradycardia. Fenoldopam treatment significantly reduced fasting but not fed blood glucose levels and lowered the blood pressure in diabetic animals. Significant change was not observed in insulin, cholesterol, triglyceride levels. Diabetic animals showed increase in AUCglucose and decrease in AUCinsulin during oral glucose tolerance test. Fenoldopam treatment did not significantly change these values in diabetic animals. STZ produced increase in serum urea, creatinine and blood urea nitrogen. Diuresis and urinary sodium retention was observed in diabetic animals. Renal hypertrophy was observed as seen from increased kidney weight/body weight ratio and increased total RNA content as well as decreased total DNA content. Fenoldopam treatment significantly lowered serum urea, creatinine and blood urea nitrogen. Urinary sodium retention was significantly reduced and renal hypertrophy was prevented with fenoldopam treatment as seen from the improved kidney weight/body weight ratio. Fenoldopam treatment significantly prevented reduction in total DNA content and increase in total RNA content further substantiating reduced renal hypertrophy. Our data suggest that STZ induced diabetes is associated with renal dysfunctions and fenoldopam treatment could be beneficial in a condition where diabetes mellitus co-exists with hypertension and compromised renal function. PMID- 11883794 TI - Communication. PMID- 11883795 TI - The circle of care: establishing a palliative care service in a long-term care facility. PMID- 11883793 TI - Daily exercise reduces measures of heart rate and blood pressure variability in hypertensive rats. AB - This study was designed to test the hypothesis that daily spontaneous running (DSR) reduces measures of heart rate and blood pressure variability in spontaneously hypertensive rats (SHR). After 8 weeks of DSR or sedentary control, rats were chronically instrumented with arterial catheters. Daily exercise reduced most measures of heart rate (HR) and blood pressure variability. Specifically DSR decreased heart rate, Low Frequency Power (LF: 0.19-0.61 Hz), and Low Frequency/High Frequency (HF: 1.2-2.5 Hz) ratio of HR. Furthermore, Total Power (TP), LF power, and LF/HF ratio of systolic blood pressure were reduced by daily spontaneous running. Finally, TP, LF and HF powers and LF/HF ratios of diastolic blood pressure were reduced by daily spontaneous running. These data demonstrate that daily exercise reduces sympathetic activity and possibly increases cardiac reserve in hypertensive animals. PMID- 11883796 TI - Should we fear the pain relief promotion act? PMID- 11883797 TI - The more things change... PMID- 11883798 TI - Ask the patient: a semi-structured interview study of quality of life in advanced cancer. AB - The purpose of this exploratory study was to identify important aspects of quality of life for patients with advanced cancer Thirty patients with advanced cancer were interviewed in this qualitative study using a 23-question, face-to face format. Questions were asked about affective, global, social, and power domains. Tape-recorded interviews were analyzed using computer data analysis software. Results indicated that patients expressed more positive feelings about their quality of life than expected. Gender differences were noted in responses in the affective, social, and power domains. Family was identified as an important component of quality of life. PMID- 11883799 TI - Palliative care and hospice: one approach. AB - The establishment of the first department of pain medicine and palliative care in a medical center in the United States is a significant marker in the development and impact of the palliative care movement. The integration of Jacob Perlow Hospice into this department is a milestone in the continued evolution of the hospice movement in the United States, and suggests that an interrelationship and interdependence between palliative care and hospice is one of the characteristics of change for the future. PMID- 11883800 TI - Supportive care of the dying: A coalition for compassionate care--Conducting an organizational assessment. PMID- 11883801 TI - Educational needs of home caregivers of terminally ill patients: literature review. AB - Hospice care continues to be a rapidly growing philosophy of care at the end of life. One of the fundamental principles of hospice is the role of a primary caregiver to provide for the needs of the terminally ill loved one. Typically, a spouse, adult child, sibling, close friend, or significant other fills this role. Usually, the caregiver has no formal training in caregiving and is learning all aspects of providing for all the needs of the patient. This is an awesome responsibility and quite overwhelming for most caregivers. A research synthesis was completed to assess the educational needs of caregivers of terminally ill patients. The computerized literature search of several databases found very few studies on the educational needs of caregivers. The few studies that have been done have used both quantitative and qualitative approaches, using a variety of assessment tools, looking at several different populations, over varied time periods. In spite of the variety of methods and populations studied, all the studies came to similar conclusions. The needs of the studied caregivers fall into three main categories: needing information on meeting the physical needs of the patients, community resources, and the patient's illness. The findings provide a basis for further research to build a comprehensive educational program that maximizes the role of the caregivers of terminally ill patients. PMID- 11883802 TI - Dyspnea assessment and management in hospice patients with pulmonary disorders. AB - Accurate assessments and appropriate management of dyspnea are essential to provide improved quality of life for hospice patients. This study describes methods of assessing dyspnea and interventions used to manage dyspnea in 72 hospice patients with end-stage lung disease or lung cancer. The mean age of the sample was 72.46 years old and the majority was white (80 percent) and male (62 percent). Paired t-tests were used to compare mean scores on admission and near death for dyspnea severity, Karnofsky functional status, pain, and Mini-Mental Status scores. Results showed significant decline in functional and cognitive status, but no significant changes in dyspnea severity and pain. Dyspnea was often assessed subjectively with observational methods only. Use of inhalants, oxygen, positioning, steroids, and oral opioids were the most frequent therapies for dyspnea. Relaxation, guided imagery, and other complementary therapies were rarely used (five percent or less). Measurement of dyspnea needs to be done frequently by using standardized instruments to assess severity and degree of symptom distress as well as the effects of treatment. Clinical trials are needed to determine which dyspnea interventions are most effective in terminally ill patients. Guidelines such as those developed for pain management are needed for effectively managing dyspnea. PMID- 11883804 TI - Spiritual "signs and symptoms": toward an expanded understanding of dying. PMID- 11883803 TI - Continuous infusion sufentanil for malignant pain: a case report. PMID- 11883806 TI - Whose "dignity" are we talking about? PMID- 11883805 TI - Building the ship of death. PMID- 11883807 TI - Wolf in sheep's clothing? PMID- 11883808 TI - Child health in an urbanizing world. AB - The aim of this study is to document and comment on the effects of urbanization on child health, internationally, using published reports and the author's personal experience. Urbanization is having profound effects on the health and well-being of infants and children in industrialized and developing countries. This will affect generations into the future. The changes are not confined to cities and large towns; they rapidly influence transitional societies in remote and rural areas, because globalization is changing infant feeding practices and children's diets and lifestyles. In developing countries, overcrowding and environmental pollution are massive problems made worse by undernutrition and infections, particularly respiratory and diarrhoeal diseases. In developed societies there are many other problems, e.g. injuries, poisonings, violence, drug abuse, exposure to industrial and atmospheric pollutants, including pesticides, sexually transmissible diseases, and "lifestyle", diseases including obesity and cardiovascular disease risk. There is an urgent need for paediatricians, health planners, policy-makers, governments and the community to understand these issues and work towards minimizing their harmful effects on children. CONCLUSION: Urbanization has profound effects on child health, globally; these must be recognised so that harmful influences of urbanization can be reduced for the benefit of all children. PMID- 11883809 TI - Presence of the genetic marker for Gilbert syndrome is associated with increased level and duration of neonatal jaundice. PMID- 11883810 TI - Rate of seizures in children with shigellosis. PMID- 11883811 TI - Child health in an urbanized world. PMID- 11883812 TI - Coeliac families. PMID- 11883813 TI - Is lactate a reliable indicator of tissue hypoxia in the neonatal period? PMID- 11883814 TI - Coeliac disease is associated with intrauterine growth and neonatal infections. AB - To investigate whether factors in the fetal or neonatal period influence the risk of later development of coeliac disease we conducted a population-based register study. The Swedish Medical Birth Register was linked with the Hospital Discharge Register and identified 3392 singleton infants born in the period 1987-97 who developed coeliac disease. Perinatal data for these children were compared with all children born in these years. Exposure variables: Maternal age, parity and smoking habits in early pregnancy, preeclampsia, pregnancy duration and birthweight, birthweight by gestational week, Apgar score, neonatal icterus, neonatal infections, maternal-fetal blood group incompatibility, exchange transfusion, phototherapy. Odds ratios and test-based confidence intervals were calculated. Analyses were made with stratification for year of birth and other risk factors. The risk of developing coeliac disease decreased with maternal age and was lower in first-born than in second-born children. Maternal smoking in early pregnancy was a weak risk factor, as was low birthweight. The most evident risk factors were being exposed to neonatal infections (OR = 1.52, confidence limits 1.19: 1.95) and being small for gestational age (OR = 1.45, confidence limits 1.20; 1.75). These risk factors were independent of each other. CONCLUSIONS: We have demonstrated that the intrauterine environment, mainly as mirrored by a low birthweight for gestational age and, independently, neonatal infection diagnosis, is associated with the risk of developing coeliac disease, supporting the idea of a multifactorial aetiology of the disease. PMID- 11883815 TI - Tissue transglutaminase autoantibodies and human leucocyte antigen in Down's syndrome patients with coeliac disease. AB - The association between autoantibodies against tissue transglutaminase (tTG) and human leucocyte antigen (HLA)-DQB1 alleles was tested in Down's syndrome (DS) patients with and without coeliac disease (CD). Immunoglobulin A (IgA) and G (IgG) anti-tTG were measured in radioligand binding assays and compared with conventionally analysed IgA antibodies against gliadin (AGA) and IgA autoantibodies against endomysium (EMA) in 48 DS patients. HLA-DQB1 typing was carried out by polymerase chain reaction and hybridization with allele-specific probes in 41/48 patients. Both IgA-tTG and IgG-tTG, as well as EMA, were detected in 7/48 and AGA in 15/48 patients. Intestinal biopsy showed histopathological changes consistent with CD in 9/16 patients. HLA-DQB1 typing, available for 8/9 patients with and for 33/39 without CD, demonstrated that 5/8 with CD had DQB1*02 compared with 7/33 of those without (p = 0.0345). In patients with anti-tTG, 5/6 had the DQB1*02 allele compared with 7/35 of those without (p = 0.0053). CONCLUSIONS: Anti-tTG are HLA-DQB1*02-associated autoantibodies which together could be useful screening tests for silent CD in DS patients. In patients with gastrointestinal symptoms or clinical signs of malabsorption, anti-tTG should be combined with AGA to detect other forms of enteropathies and CD. PMID- 11883816 TI - Nature and extent of gastric lesions in symptomatic Chilean children with Helicobacter pylori-associated gastritis. AB - Chile has one of the highest rates of gastric cancer in the world and most children and adolescents in the country are colonized by Helicobacter pylori. This study assessed the nature and extent of the gastric lesions in 73 consecutive patients aged 5-17 y, referred for upper gastrointestinal endoscopy. Their H. pylori-associated gastric pathology was characterized and these data were compared with their sociodemographic status. Endoscopic assessment was normal in 43 patients while in 30 there was a variety of mucosal lesions. Sixty patients (83%) had histological chronic gastritis of the antrum and in 45 (63%) the lesions also involved the gastric corpus; 90% of patients with chronic gastritis were colonized by H. pylori. Although most of these patients had epithelial erosions and dedifferentiation of the pit epithelium, atrophy and metaplasia were not found. Patients' socioeconomic status was inversely correlated with their rate of colonization by H. pylori (p < 0.005), the frequency of gastric lesions on endoscopy (p < 0.01) and the frequency of involvement of antral and corpus mucosa by chronic gastritis (p < 0.002). This latter feature was positively correlated with age (p < 0.001). CONCLUSION: This study shows a high frequency of extensive lesions of H. pylori-associated chronic gastritis in young Chilean patients. This histological picture is consistent with the hypothesis of a H. pylori-associated progressive gastric pathology which may represent a major factor in the high local rate of gastric cancer. PMID- 11883817 TI - Faecal calprotectin levels in infants with infantile colic, healthy infants, children with inflammatory bowel disease, children with recurrent abdominal pain and healthy children. AB - This study investigated faecal calprotectin concentration, a measure of intestinal inflammation, in infants and children with abdominal pain. Faecal calprotectin was measured by an enzyme-linked immunosorbent assay kit in spot stool samples in 76 infants with typical infantile colic, 7 infants with transient lactose intolerance and 27 healthy infants. All infants were 2-10 wk of age. In addition, 19 children with recurrent abdominal pain (RAP; mean age 11.5 y), 17 with inflammatory bowel disease (IBD; mean age 11.1 y; 10 had Crohn's disease and 7 ulcerative colitis) and 24 healthy children (mean age 5.3 y) were studied. In infants with infantile colic the mean faecal calprotectin concentration was not different from that in healthy infants (278 +/- 105 vs 277 +/- 109 mg kg(-1), p = 0.97) or in infants with transient lactose intolerance (300.3 +/- 124 mg kg(-1), p = 0.60). The calprotectin level was similar in boys and girls and fell significantly with age (p = 0.04). Children with IBD had faecal calprotectin levels (293 +/- 218 mg kg(-1)) much higher than healthy children (40 +/- 28 mg kg(-1), p < 0.0001) and children with RAP without identified organic disease (18 +/- 24 mg kg(-1), p < 0.0001). CONCLUSION: Faecal calprotectin may differentiate between functional abdominal pain and IBD in school-aged children. In young infants high faecal calprotectin levels are normal. PMID- 11883818 TI - Folic acid supplementation on red kidney bean-induced diarrhoea and enteric bacterial translocation into mesenteric lymph nodes in rats: a pilot study. AB - Deaths following childhood diarrhoea, a major health problem in developing countries, are often associated with malnutrition and septicaemic complications. Folic acid has been used in the treatment of acute and chronic diarrhoea in the tropics. Using a rat model, we evaluated the protective effect of large doses of folic acid on diarrhoea, small intestinal bacterial overgrowth and translocation of enteric bacteria into mesenteric lymph nodes induced by a raw red kidney bean based diet containing lectin (phytohemagglutinin). Long-Evans rats in 2 groups of 5 each (60 g to 70 g in weight, 28 d old) were used. All 10 rats, individually kept in metabolic cages, received a raw red kidney bean-based diet for 10 d, and 5 of them also received a daily folic acid supplement (160 microg/g feed) both during and for 10 d before the experiment. The faecal weight was measured and a quantitative aerobic bacterial culture of the small intestinal mucosal scrapings and of the mesenteric lymph nodes was made. Folic acid supplementation did not reduce faecal output nor did it prevent loss of body weight associated with lectin-induced diarrhoea. However, the mean total count of enteric bacteria translocated to the mesenteric lymph nodes was significantly reduced in the supplemented rats (1.27 +/- 0.61 vs 2.66 +/- 0.84, p = 0.028) and a trend towards reduced bacterial count in the small intestinal mucosal scrapings (0.40 +/- 0.89 vs 1.42 +/- 1.31, p = 0.16) was documented. A significant positive correlation was also seen between the bacterial count in the jejunal mucosal scrapings and in the mesenteric lymph nodes. CONCLUSION: Although large-dose folic acid supplementation did not prevent diarrhoea and malnutrition induced by a lectin based diet, it substantially reduced the count of enteric bacteria translocated into the mesenteric lymph nodes and showed a trend towards a reduction in indigenous bacteria adhering to jejunal mucosa. These findings could be of relevance in the prevention of septicaemic complications following many clinical conditions, including diarrhoea with malnutrition in children known to have bacteraemic and septicaemic complications. PMID- 11883819 TI - Antibiotic treatment for five days is effective in children with acute cystitis. AB - Short courses of antibiotics are often recommended to treat children with acute cystitis despite lack of firm evidence to support such management. The aim of this study therefore was to analyse the short-term outcome of such treatment. The retrospective analysis included 300 children (252F, 48M) fulfilling the criteria of first-time acute cystitis and managed according to a protocol recommending 5 d treatment. In 214 (71%) the treatment was given according to the protocol and in the others for 7 or 10 d. Nitrofurantoin was used in 150 (50%) and trimethoprim without or with sulfonamide in 129 (43%). The short-term results were excellent with 96% of the children being free from symptoms at the first follow-up visit after a median of 6 d. Only 2 girls had persisting bacteriuria and thus the frequency of bacteriological treatment failure was 1%. Recurrence within 30 d occurred in 4 girls (2%). CONCLUSION: A 5 d treatment with antibiotics is adequate in children with acute cystitis. Routine follow-up visits after a first acute cystitis may not be necessary, providing that the bacteria causing the infection are sensitive to the prescribed antibiotic and that there is no history of defective bladder or bowel emptying. PMID- 11883820 TI - Rickettsia typhi infection in childhood. AB - Rickettsia typhi infection (murine typhus) is generally underdiagnosed in childhood, as clinical presentations are often non-specific. We present the manifestations in nine children hospitalized in the Department of Paediatrics of the University Hospital, Heraklion, Crete, over a 3-y period from 1998 to 2000. Titres > 1:400 for IgM and >1:960 for IgG and/or a fourfold increase in a second sample were considered strongly suggestive of acute infection. Children presented with prolonged fever, hepatosplenomegaly and lymphadenopathy. Five children presented with a rash. Unusual manifestations included aseptic meningitis and Kawasaki-like presentation. Laboratory findings included anaemia, leucopenia, and thrombocytopenia. Three children were treated with appropriate antibiotic regimens and all nine had a complete recovery. CONCLUSION: Rickettsia typhi infection should be considered in the differential diagnosis of children residing in or returning from Southern Europe countries who present with prolonged fever, rash and lymphadenopathy. PMID- 11883821 TI - Visceral childhood leishmaniasis in Turkey. AB - Between 1981 and 2001, we retrospectively analysed 40 cases of visceral leishmaniasis (VL) admitted to the Paediatric Infection Unit of Ondokuz Mayis University Hospital, in the middle Black Sea region of Turkey. Median age at presentation was 3 y. Fever and splenomegaly were found in all patients. Bone marrow smear examination resulted in the diagnosis of VL in 95% of cases. All patients were treated initially with meglumine antimonate and 95% of them were cured with this therapy. The remaining patients were cured with liposomal amphotericin B. CONCLUSIONS: VL should be considered in patients with fever and splenomegaly, particularly those residing in the Mediterranean region. Meglumine antimonate seems to be the first choice of treatment in childhood. PMID- 11883822 TI - Cardiopulmonary exercise parameters in children with atrial septal defect and increased pulmonary blood flow: short-term effects of defect closure. AB - Markedly increased pulmonary blood flow because of a relevant atrial septal defect (ASD) leads to impaired cardiopulmonary function during maximum exercise in adults. No comparative preoperative and postoperative data are available on the short-term effects of shunt closure on cardiorespiratory function at peak exercise in children. Pulmonary function testing at rest and cardiopulmonary exercise testing together with haemodynamic assessment was done prospectively in children with an ASD preoperatively and again after full recovery at 3-4 mo postoperatively and compared with a matched normal population. Sixteen children, aged 6.8-16.1 y, with a defect of 8-23 mm (median 15 mm) and a pulmonary/systemic flow ratio of 1.5-3.5 (median 2.2) were tested and compared with 15 healthy children. Preoperatively, baseline pulmonary function parameters and exercise capacity were no different from normals. At peak exercise, patients with a shunt had increased pulmonary resistance, especially of the distal airways (p = 0.04), with a significantly larger proportion of children having a paradoxical increase in total airway resistance during exercise (p < 0.05). Maximum serum lactate at peak exercise was elevated (p < 0.05) in patients. In patients, maximum oxygen uptake was impaired (p = 0.03) and remained so at repeat evaluation postoperatively. The same observation was made for chronotropic response to exercise. CONCLUSION: Cardiopulmonary exercise parameters in patients with ASD differed only slightly from those in normal children. The most important deviations were a lower maximum oxygen uptake and an increase in airway resistance at maximum exercise. PMID- 11883823 TI - Craniofacial morphology in preschool children with sleep-related breathing disorder and hypertrophy of tonsils. AB - The purpose of this study was to examine craniofacial morphology, pharyngeal airway space and hyoid bone position in preschool children with sleep-related breathing disorder associated with hypertrophy of tonsils (SBDT). Thirty-eight preschool children, mean age 4.7 y, with SBDT and with an apnoea index (AI) of 0 < AI < 5, were divided into two groups. One consisted of 15 children with sleep related breathing disorder (SBD) and more than 75% of the tonsils visible (GIII) and the other of 23 children with SBD and 25-75% of the tonsils visible (GII). The control group consisted of 31 children without ear, nose and throat disease and with GI (barely visible) tonsils. Compared with the controls, GIII children had a retrognathic mandible, a large posterior facial height, a large interincisal angle with retroclined lower incisors, a narrow pharyngeal airway space, an anterior tongue base position and a long soft palate. Compared with the controls, GII children had a large anterior lower facial height and a short nasal floor. However, like the controls, GII children did not have a retrognathic mandible. CONCLUSION: The findings show that children with SBDT display a characteristic facial appearance at an early age. Since the condition has an effect on growth, it needs to be prevented by controlling morphology and function at the preschool age. PMID- 11883824 TI - Agreement between capillary and arterial lactate in the newborn. AB - Arterial blood lactate is a reliable indicator of tissue oxygen debt and is of value in expressing the degree and prognosis of circulatory failure as a result of various diseases. Therefore, the practical issue of whether capillary lactate measurements might be of equal value was investigated in newborns. In total, 193 simultaneous measurements of capillary and arterial blood lactate concentrations were performed in 25 newborn babies with an indwelling umbilical arterial catheter. A strong linear correlation was found between capillary and arterial lactate concentration (Lcap = 1.02 Lart + 0.04; r = 0.98; p < 0.001). The mean difference was -0.08 mmol/l and the limits of agreement (+/- 2 SD) were +/- 0.69 mmol/l (-0.77 to 0.61 mmol/l). CONCLUSION: Our data show that capillary blood lactate measurements in newborn babies yield lactate concentrations equivalent to arterial measurements over a large concentration range. PMID- 11883825 TI - Genetic and clinical features of false-negative infants in a neonatal screening programme for cystic fibrosis. AB - A study was performed on the delayed diagnosis of cystic fibrosis (CF) in infants who had false-negative results in a neonatal screening programme. The genetic and clinical features of false-negative infants in this screening programme were assessed together with the efficiency of the screening procedure in the Lombardia region. In total, 774,687 newborns were screened using a two-step immunoreactive trypsinogen (IRT) (in the years 1990-1992), IRT/IRT + delF508 (1993-1998) or IRT/IRT + polymerase chain reaction (PCR) and oligonucleotide ligation assay (OLA) protocol (1998-1999). Out of 196 CF children born in the 10 y period 15 were false negative on screening (7.6%) and molecular analysis showed a high variability in the genotypes. The cystic fibrosis transmembrane regulator (CFTR) gene mutations identified were delF508, D1152H, R1066C, R334W, G542X, N1303K, F1052V, A120T, 3849 + 10kbC --> T, 2789 + 5G --> A, 5T-12TG and the novel mutation D110E. In three patients no mutation was identified after denaturing gradient gel electrophoresis of the majority of CFTR gene exons. CONCLUSION: The clinical phenotypes of CF children diagnosed by their symptoms at different ages were very mild. None of them presented with a severe lung disease. The majority of them did not seem to have been damaged by the delayed diagnosis. The combination of IRT assay plus genotype analysis (1998-1999) appears to be a more reliable method of detecting CF than IRT measurement alone or combined with only the delF508 mutation. PMID- 11883826 TI - Adverse drug events in children during hospitalization and after discharge in a Norwegian university hospital. AB - The frequency and characteristics of adverse drug events (ADEs) in children hospitalized in the paediatric department of Ullevaal University Hospital, Norway, were determined using intensive monitoring. Of 579 children treated with drugs, 28% experienced ADEs; 7% at the time of admission, 18% during hospitalization and 9% after discharge. All children treated for cancer, 19% treated with anti-infective drugs, 15% treated with antiasthmatics and 10% treated with drugs affecting the nervous system experienced ADEs. The most frequent events were gastrointestinal, CNS- and skin reactions and 19% were considered as serious. ADEs caused 6% of the admissions and 44% required interventions. Most ADEs were found by screening patient records, where physicians mostly described adverse drug events requiring interventions and nurses described less serious events. Parents reported 14% of the events, of which a majority were CNS reactions. CNS reactions may be more common than expected and observations by parents are important when investigating such reactions in children. CONCLUSIONS: ADEs, mainly gastrointestinal, CNS and skin reactions related to drugs affecting the nervous system, anti-infectives and antiasthmatics, were seen in 28% of the patients. The reporting of events by parents was a useful supplement to the screening of patient records. PMID- 11883827 TI - Brain damage markers in children. Neurobiological and clinical aspects. AB - The presence in blood of proteins normally confined to the cytoplasm of brain cells is considered peripheral evidence of brain damage. Only recently have these proteins been measured in the blood of children at risk of brain damage. To show the value and limitations of measuring these proteins, we review their biology and the adult literature that has correlated the blood concentrations of these proteins with lesion size and dysfunction. CONCLUSION: We conclude that brain damage markers will increasingly be measured in the blood of newborns and other children at risk of brain damage. PMID- 11883828 TI - Orbital lymphangioma with non-contiguous cerebral arteriovenous malformation, manifesting with thrombocytopenia (Kasabach-Merritt syndrome) and intracerebral hemorrhage. AB - This study describes the first reported case in a preterm infant of an orbital lymphangioma with non-contiguous cerebral arteriovenous malformation, manifesting with thrombocytopenia (Kasabach-Merritt syndrome) and intracerebral hemorrhage. CONCLUSION: Neonates presenting with orbital lymphangiomas should undergo radiological investigations of the lesion and a detailed cerebral evaluation for associated arteriovenous developmental anomalies. PMID- 11883829 TI - Trigeminal schwannomas: removal of dumbbell-shaped tumors through the expanded Meckel cave and outcomes of cranial nerve function. AB - OBJECT: As in patients with vestibular schwannomas, advances in surgical procedures have markedly improved outcomes in patients with trigeminal schwannomas. In this article the authors address the function of cranial nerves in a series of patients with trigeminal schwannomas that were treated with gross total surgical removal. The authors emphasize a technique they use to remove a dumbbell-shaped tumor through the expanded Meckel cave, and discuss the advantage of the extradural zygomatic middle fossa approach for total removal of tumor and preservation or improvement of cranial nerve function. METHODS: Within an 11-year period (1989-2000), 25 patients (14 female and 11 male patients with a mean age of 44.4 years) with benign trigeminal schwannomas were surgically treated by the senior author (O.A.) with the aim of total removal of the tumor. Three patients had undergone previous surgery elsewhere. Trigeminal nerve dysfunction was present in all but two patients. Abducent nerve paresis was present in 40%. The approach in each patient was selected according to the location and size of the lesion. Nineteen tumors were dumbbell shaped and extended into both middle and posterior fossae. All 25 tumors involved the cavernous sinus. The zygomatic middle fossa approach was particularly useful and was used in 14 patients. The mean follow-up period was 33.12 months. In patients who had not undergone previous surgery, the preoperative trigeminal sensory deficit improved in 44%, facial pain decreased in 73%, and trigeminal motor deficit improved in 80%. Among patients with preoperative abducent nerve paresis, recovery was attained in 63%. Three patients (12%) experienced a persistent new or worse cranial nerve function postoperatively. Fifth nerve sensory deficit persisted in one of these patients, sensory and motor dysfunction in another, and motor trigeminal weakness in the third patient. In all patients a good surgical outcome was achieved. One patient died 2 years after treatment from an unrelated cause. In three patients the tumors recurred after an average of 22.3 months. CONCLUSIONS: Preservation or improvement of cranial nerve function can be achieved through total removal of a trigeminal schwannoma, and skull base approaches are better suited to achieving this goal. The zygomatic middle fossa approach is particularly helpful and safe. It allows extradural tumor removal from the cavernous sinus, the infratemporal fossa, and the posterior fossa through the expanded Meckel cave. PMID- 11883830 TI - Individual variations in the sulcal anatomy of the basal temporal lobe and its relevance for epilepsy surgery: an anatomical study performed using magnetic resonance imaging. AB - OBJECT: The concept of selective amygdalohippocampectomy is based on pathophysiological insights into the epileptogenicity of the hippocampal region and the definition of the clinical syndrome of mesial temporal lobe epilepsy (TLE). High-resolution magnetic resonance (MR) imaging allows correlation of the site of histologically conspicuous tissue with anatomical structure. The highly variable sulcal pattern of the basal temporal lobe, however, definitely complicates the morphometric analysis of histomorphologically defined subdivisions of the hippocampal region. The goal of this study was to define individual variations in the sulcal anatomy on the basis of preoperative MR images obtained in patients suffering from TLE. METHODS: The authors analyzed coronal MR images obtained in 50 patients for the presence of and intrinsic relationships among the rhinal, collateral, and occipitotemporal sulci. The surface relief of consecutive sections of 100 temporal lobes was graphically outlined and the resulting maps were used for visual analysis. The sulci were characterized by measurement of their depth, distance to the temporal horn, and laterality. The anatomical measurements and frequencies of sulcal patterns were assessed for statistical correlation with patients' histories and the lateralization of the seizure focus. CONCLUSIONS: Statistical assessment shows that patient sex is a significant factor in sulcal patterns. Anatomical measurements are significantly decreased on the side of the seizure origin, which relates to loss of white matter, a known morphological abnormality associated with TLE. Magnetic resonance imaging allows for accurate preoperative knowledge of individual sulcal patterns and facilitates intraoperative orientation to anatomical landmarks. PMID- 11883831 TI - Endovascular treatment of giant and large intracranial aneurysms by using a combination of stent placement and liquid polymer injection. AB - OBJECT: The aim of this study was to test the feasibility, safety, and efficacy of a new endovascular method for the treatment of giant intracranial aneurysms. This new method consists of combining a metallic stent with a liquid polymer; the stent is first placed across the neck of the aneurysm to reconstruct a tubular arterial lumen, followed by obliteration of the fundus of the aneurysm with an ethyl vinyl alcohol polymer. During its injection, the liquid polymer is contained within the aneurysm by temporarily inflating an occlusion balloon in the parent artery. METHODS: Eleven patients harboring a giant aneurysm were successfully treated using this procedure. All aneurysms were excluded from the circulation, with preservation of the parent artery. In nine of the 11 patients, the 6-month follow-up angiogram demonstrated no recanalization of the aneurysm. In one patient who had a giant and partially clotted internal carotid artery bifurcation aneurysm, the follow-up angiogram demonstrated minimal recanalization. The complications in this series of patients included one death and one case of transient hemiparesis caused by watershed ischemia. CONCLUSIONS: The initial anatomical results and the clinical outcome in this small series of patients are very encouraging. The mortality and morbidity rates associated with this new endovascular treatment are superior to those associated with surgical clipping of giant aneurysms. PMID- 11883832 TI - Spontaneous spinal cerebrospinal fluid leaks and minor skeletal features of Marfan syndrome: a microfibrillopathy. AB - OBJECT: Spontaneous spinal cerebrospinal fluid (CSF) leaks are increasingly recognized as a cause of postural headaches. The authors examined a group of patients suffering from spontaneous spinal CSF leaks who also had minor skeletal features of Marfan syndrome for abnormalities of fibrillin-containing microfibrils. METHODS: Patients with spontaneous CSF leaks were evaluated for the clinical characteristics of connective tissue disorders. Skin biopsies were obtained in three patients with skeletal manifestations that constitute part of the Marfan syndrome phenotype. Cultured fibroblasts were studied for fibrillin-1 synthesis and incorporation into the extracellular matrix (ECM) by performing quantitative metabolic labeling and immunohistochemical analysis. Among 20 consecutive patients found to have spinal CSF leaks, four (20%) exhibited minor skeletal features of Marfan syndrome, but lacked any ocular or cardiovascular abnormalities. The mean age of these patients (30 years) was lower than that of the 16 patients without skeletal abnormalities (44 years; p = 0.01). Abnormalities in fibrillin-1 metabolism and immunostaining were detected in all three patients with the skeletal abnormalities who underwent examination, but not in a control patient without these skeletal manifestations. CONCLUSIONS: Twenty percent of patients who experience spontaneous spinal CSF leaks have minor skeletal features of Marfan syndrome. The authors demonstrated abnormalities in fibrillin-1 protein deposition in all patients examined, but only one person was found to have a fibrillin-1 abnormality typically found in classic Marfan syndrome. The results indicate that there is a heterogeneous involvement of other components of ECM microfibrils at the basis of this cerebrospinal manifestation. In addition, the authors identified a connective-tissue etiological factor in a group of disorders not previously classified as such. PMID- 11883833 TI - Stent placement for the treatment of occlusive atherosclerotic carotid artery disease in patients with concomitant coronary artery disease. AB - OBJECT: The authors report their experience with carotid artery stent placement (CASP) in patients with concomitant carotid artery (CA) and coronary artery (CorA) diseases. METHODS: In a review of 320 consecutive patients who underwent CASP, the authors identified 49 with severe CorA disease in addition to significant CA stenosis, who had undergone CASP before planned CorA bypass grafting (CorABG). The average age of these 49 patients was 68 years. In 39 patients (80%) the New York Heart Association functional classification grade was IV and in 10 the grade was III. In 26 patients 50% or greater stenosis of the left main CorA was found. Seventeen patients (35%) suffered from either significant hemodynamic contralateral CA stenosis (> 60% stenosis; eight patients) or contralateral CA occlusion (nine patients). Sixteen patients (33%) had symptomatic CA disease. No cerebrovascular events occurred during CorABG. Four patients (8%) died of cardiac arrest and one patient (2%) suffered a major stroke within 30 days after the CorABG procedure. No patient experienced clinically significant recurrent CA stenosis during the study period (average clinical follow-up period 27 months). CONCLUSIONS: Carotid artery stent placement should be considered as an alternative for the management of concomitant CA and CorA diseases. These preliminary results support the feasibility and durability of CASP in the population studied. PMID- 11883834 TI - Seasonal variation in the incidence of aneurysmal subarachnoid hemorrhage in hospital- and community-based studies. AB - OBJECT: The aim of this study was to examine seasonal variations in the onset of aneurysmal subarachnoid hemorrhage (SAH) in one hospital-based and one community based patient series. METHODS: The study population consisted of 941 patients with aneurysmal SAH who were admitted to Shimane Prefectural Central Hospital and 358 patients who were treated in Izumo City, Japan. When investigated as a whole, no significant seasonal variations were found in either population; however, in both series, statistically significant seasonal trends, with a peak in winter and a nadir in summer, were found among patients aged 59 years or younger (p < 0.05 for the hospital-based series and p < 0.005 for the community-based series), but not among those aged 60 years or older, regardless of sex. In the hospital-based series, seasonal variations were most apparent at certain times of day, with significant variations observed between 8:00 a.m. and 10:00 a.m. or 8:00 a.m. and noon (p < 0.001 and p < 0.005, respectively), regardless of patient age, and between 4:00 p.m. and 6:00 p.m. for patients aged 59 years or younger (p < 0.05). Consequently, seasonal variations were significant during daytime hours (between 8:00 a.m. and 8:00 p.m., p < 0.005) but not during the night (between 8:00 p.m. and 8:00 a.m.). Similar tendencies were found in the community-based series. Among patients aged 59 years or younger who had no risk factors for SAH, these seasonal variations were significant in both series. In patients with untreated hypertension, who were current smokers and daily alcohol drinkers, however, no significant patterns were observed in either series, even among younger patients. CONCLUSIONS: In both hospital- and community-based studies, aneurysmal SAH appears to undergo seasonal variation, with a peak in winter and a nadir in summer. This seasonal pattern may be derived mainly from the occurrence of SAH in the morning, but may also be modified by patient age and SAH risk factors, resulting in the masking of significant seasonal patterns when all patients are considered together. PMID- 11883835 TI - Magnesium sulfate therapy after aneurysmal subarachnoid hemorrhage. AB - OBJECT: Vasospasm remains a significant source of neurological morbidity and mortality following aneurysmal subarachnoid hemorrhage (SAH), despite advances in current medical, surgical, and endovascular therapies. Magnesium sulfate therapy has been demonstrated to be both safe and effective in preventing neurological complications in obstetrical patients with eclampsia. Evidence obtained using experimental models of brain injury, cerebral ischemia, and SAH indicate that Mg may also have a role as a neuroprotective agent. The authors hypothesize that MgSO4 therapy is safe, feasible, and has a beneficial effect on vasospasm and, ultimately, on neurological outcome following aneurysmal SAH. METHODS: A prospective randomized single-blind clinical trial of high-dose MgSO4 therapy following aneurysmal SAH (Hunt and Hess Grades II-IV) was performed in 40 patients, who were enrolled within 72 hours following SAH and given intravenous MgSO4 or control solution for 10 days. Serum Mg++ levels were maintained in the 4 to 5.5 mg/dl range throughout the treatment period. Clinical management principles were the same between groups (including early use of surgery or endovascular treatment, followed by aggressive vasospasm prophylaxis and treatment). Daily transcranial Doppler (TCD) ultrasonographic recordings were obtained, and clinical outcomes were measured using the Glasgow Outcome Scale (GOS). The patients' GOS scores and the TCD recordings were analyzed using the independent t-test. Forty patients were enrolled in the study: 20 (15 female and five male patients) received treatment and 20 (11 female and nine male patients) comprised a control group. The mean ages of the patients in these groups were 46 and 51, respectively, and the mean clinical Hunt and Hess grades were 2.6 +/- 0.68 in the MgSO4 treatment group and 2.3 +/- 0.73 in the control group (mean +/- standard deviation [SD], p = 0.87). Fisher grades were similar in both groups. Mean middle cerebral artery velocities were 93 +/- 27 cm/second in MgSO4-treated patients and 102 +/- 34 cm/second in the control group (mean +/- SD, p = 0.41). Symptomatic vasospasm, confirmed by angiography, occurred in six of 20 patients receiving MgSO4 and in five of 16 patients receiving placebo. Mean GOS scores were 3.8 +/- 1.6 and 3.6 +/- 1.5 (mean +/- SD, p = 0.74) in the treatment and control groups, respectively. Significant adverse effects from treatment with MgSO4 did not occur. CONCLUSIONS: Administration of high-dose MgSO4 following aneurysmal SAH is safe, and steady Mg++ levels in the range of 4 to 5.5 mg/dl are easily maintained. This treatment does not interfere with neurological assessment, administration of anesthesia during surgery, or other aspects of clinical care. We observed a trend in which a higher percentage of patients obtained GOS scores of 4 or 5 in the group treated with MgSO4, but the trend did not reach a statistically significant level. A larger study is needed to evaluate this trend further. PMID- 11883836 TI - Intraoperative complications in aneurysm surgery: a prospective national study. AB - OBJECT: With increasing use of endovascular procedures, the number of aneurysms treated surgically will decline. In this study the authors review complications related to the surgical treatment of aneurysms and address the issue of maintaining quality standards on a national level. METHODS: A prospective, nonselected amalgamation of every aneurysm case treated in five of six neurosurgical centers in Sweden during 1 calendar year was undertaken (422 patients; 7.4 persons/100,000 population/year). The treatment protocols at these institutions were very similar. Outcome was assessed using clinical end points. In this series, 84.1% of the patients underwent surgery, and intraoperative complications occurred in 30% of these procedures. Poor outcome from technical complications was seen in 7.9% of the surgically treated patients. Intraoperative aneurysm rupture accounted for 60% and branch sacrifice for 12% of all technical difficulties. Although these complications were significantly related to aneurysm base geometry and the competence of the surgeon, problems still occurred apparently at random and also in the best of hands (17%). The temporary mean occlusion time in the patients who suffered intraoperative aneurysm rupture was twice as long as the temporary arrest of blood flow performed to aid dissection. CONCLUSIONS: The results obtained in this series closely reflect the overall management results of this disease and support the conclusion that surgical complications causing a poor outcome can be estimated on a large population-based scale. Intraoperative aneurysm rupture was the most common and most devastating technical complication that occurred. Support was found for a more liberal use of temporary clips early during dissection, regardless of the experience of the surgeon. Temporary regional interruption of arterial blood flow should be a routine method for aneurysm surgery on an everyday basis. A random occurrence of difficult intraoperative problems was clearly shown, and this factor of unpredictability, which is present in any preoperative assessment of risk, strengthens the case for recommending neuroprotection as a routine adjunct to virtually every aneurysm operation, regardless of the surgeon's experience. PMID- 11883837 TI - Brachial plexus neurotization with donor phrenic nerves and its effect on pulmonary function. AB - OBJECT: To examine possible side effects of neurotizations in which the phrenic nerve was used, pulmonary function was analyzed pre- and postoperatively in patients with brachial plexus injury and root avulsions. METHODS: Twenty-three patients with complete brachial plexus palsy underwent neurotization of the musculocutaneous nerve, with the phrenic nerve as donor material. Patients who suffered lung contusions as part of the primary injury were excluded from this study. In 12 patients (five left-sided and seven right-sided neurotizations) pre- and postoperative functional parameters were compared and additional body plethysmography was performed more than 12 months postsurgery. Of the 23, no patient experienced pulmonary problems postoperatively. Nonetheless, pulmonary functional parameters showed a vital capacity in percent of the predicted value of 9.8 +/- 6.3% (mean +/- standard deviation [SD]) in all patients examined, which was a significant reduction (p = 0.0002). In right-sided phrenic nerve transfers this reduction was significant, at 14.3 +/- 3.3% (mean +/- SD), whereas left-sided transfers showed a nonsignificant reduction of 3.6 +/- 3.5% (mean +/- SD). The observed decrease in vital capacity (VC) correlates with the maximal inspiratory pressure (Pi(max)) as an indication of clinical significance. CONCLUSIONS: When the right phrenic nerve is used as a donor in neurotization of the musculocutaneous nerve, the patient incurs a higher risk of reduced pulmonary VC. If possible, the left phrenic nerve should be preferred. The Pi(max) has to be determined preoperatively to avoid any further decrease in the already reduced pulmonary function due to the initial injury. PMID- 11883838 TI - Predictors of outcome in surgically managed patients with typical and atypical trigeminal neuralgia: comparison of results following microvascular decompression. AB - OBJECT: Microvascular decompression (MVD) has become one of the primary treatments for typical trigeminal neuralgia (TN). Not all patients with facial pain, however, suffer from the typical form of this disease; many patients who present for surgical intervention actually have atypical TN. The authors compare the results of MVD performed for typical and atypical TN at their institution. METHODS: The results of 2675 MVDs in 2264 patients were reviewed using information obtained from the department database. The authors examined immediate postoperative relief in 2003 patients with typical and 672 with atypical TN, and long-term follow-up results in patients for whom more than 5 years of follow-up data were available (969 with typical and 219 with atypical TN). Outcomes were divided into three categories: excellent, pain relief without medication; good, mild or intermittent pain controlled with low-dose medication; and poor, no or poor pain relief with large amounts of medication. The results for typical and atypical TN were compared and patient history and pain characteristics were evaluated for possible predictive factors. CONCLUSIONS: In this study, MVD for typical TN resulted in complete postoperative pain relief in 80% of patients, compared with 47% with complete relief in those with atypical TN. Significant pain relief was achieved after 97% of MVDs in patients with typical TN and after 87% of these procedures for atypical TN. When patients were followed for more than 5 years, the long-term pain relief after MVD for those with typical TN was excellent in 73% and good in an additional 7%, for an overall significant pain relief in 80% of patients. In contrast, following MVD for atypical TN, the long term results were excellent in only 35% of cases and good in an additional 16%, for overall significant pain relief in only 51%. Memorable onset and trigger points were predictive of better postoperative pain relief in both atypical and typical TN. Preoperative sensory loss was a negative predictor for good long-term results following MVD for atypical TN. PMID- 11883839 TI - Mechanism of trigeminal neuralgia: an ultrastructural analysis of trigeminal root specimens obtained during microvascular decompression surgery. AB - OBJECT: Recent progress in the understanding of abnormal electrical behavior in injured sensory neurons motivated an examination, at the ultrastructural level, of trigeminal roots of patients with trigeminal neuralgia (TN). METHODS: In 12 patients biopsy specimens of trigeminal root were obtained during surgery for microvascular decompression. Pathological changes in tissue included axonopathy and axonal loss, demyelination, a range of less severe myelin abnormalities (dysmyelination), residual myelin debris, and the presence of excess collagen, including condensed collagen masses in two cases. Within zones of demyelination, groups of axons were often closely apposed without an intervening glial process. Pathological characteristics of nerve fibers were clearly graded with the degrees of root compression noted at operation. Pain also occurred, however, in some patients who did not appear to have a severe compressive injury. CONCLUSIONS: Findings were consistent with the ignition hypothesis of TN. This model can be used to explain the major positive and negative symptoms of TN by axonopathy induced changes in the electrical excitability of afferent axons in the trigeminal root and of neuronal somata in the trigeminal ganglion. The key pathophysiological changes include ectopic impulse discharge, spontaneous and triggered afterdischarge, and crossexcitation among neighboring afferents. PMID- 11883840 TI - Gamma surgery for melanoma metastases in the brain. AB - OBJECT: The aim of this study was to evaluate the usefulness and limitations of gamma surgery (GS) in the treatment of brain metastases from melanoma. METHODS: Imaging and clinical outcomes in 45 patients treated for 92 brain metastases from melanoma between October 1989 and October 1999 were retrospectively analyzed. Follow-up imaging studies were available in 35 patients with 66 treated lesions. Twenty-four percent of the lesions disappeared, 35% shrank, 23% remained unchanged, and 18% increased in size. No undue radiation-induced changes were observed in the surrounding brain. Clinical data were available in all patients. No deaths or neurological morbidity related to GS was observed. The median survival time, calculated using the Kaplan-Meier method, was 10.4 months from the time of GS. In both univariate and multivariate Cox regression analyses, a single brain lesion and lack of visceral metastases were statistically predictive of a better prognosis. Six of eight patients with solitary metastasis (that is, a single brain metastasis with no primary visceral tumor) were still alive at the close of the study, none of them with disease progression, with a follow-up period ranging between 14 and 82 months. Sixteen patients in this series received adjunctive whole-brain radiation therapy, which had no impact on their survival time or local and distant control of the brain disease. CONCLUSIONS: Gamma surgery is effective in treating melanoma metastases in the brain. It appears that the radiobiology of a single high dose overcomes the radioresistance barrier, yielding better results than fractionated radiation. PMID- 11883841 TI - Surgical management of cerebral metastases from melanoma: outcome in 147 patients treated at a single institution over two decades. AB - OBJECT: The aim of this study was to review the outcome of patients who underwent surgery for treatment of cerebral metastatic melanoma. METHODS: A retrospective analysis was performed in 147 patients with cerebral metastases from melanoma who were treated surgically at a single institution between 1979 and 1999. Almost all patients underwent postoperative wholebrain radiation therapy. The mean patient age was 53 years (range 17-76 years); 69% of patients were male. A single cerebral metastasis was identified in 84% of patients, although 56% had synchronous extracranial metastases. The 30-day postoperative mortality rate was 2% and neurological symptoms resolved or improved in 78% of patients. Recurrence of intracerebral disease was seen in 55% of patients and 26% died of intracerebral metastases. Twenty-four patients underwent reoperation for recurrent cerebral disease. The median survival duration from the time of surgery for all patients was 8.5 months; the 3- and 5-year survival rates were 9% and 5%, respectively. Factors that significantly influenced survival on univariate analysis were the number of cerebral metastases (p = 0.015), a macroscopically complete excision (p < 0.05), and reoperation for recurrence (p = 0.02). The presence of extracranial metastases did not significantly influence survival. On multivariate analysis only the number of cerebral metastases significantly affected survival (p = 0.04). CONCLUSIONS: For the majority of patients with cerebral metastases from melanoma, surgery with adjuvant radiation therapy is a treatment option that improves neurological symptoms and produces minimal morbidity. Long-term survival (> 3 years) most likely occurs in patients with a single cerebral metastasis and no demonstrable extracranial disease. Reoperation for recurrent cerebral disease may be appropriate in selected cases. PMID- 11883842 TI - Does administration of recombinant human erythropoietin attenuate the increase of S-100 protein observed in cerebrospinal fluid after experimental subarachnoid hemorrhage? AB - OBJECT: Results of recent studies indicate that erythropoietin (EPO) produces a neuroprotective effect on experimental subarachnoid hemorrhage (SAH). It has been reported that S-100 protein levels increase in cerebrospinal fluid (CSF) after SAH, providing a highly prognostic indication of unfavorable outcome. This study was conducted to validate further the findings of S-100 protein as an index of brain damage and to assess whether treatment with recombinant human EPO (rhEPO) would limit the increase of S-100 protein level in CSF following experimental SAH. METHODS: Thirty-two rabbits were each assigned to one of four groups: Group 1, control; Group 2, SAH; Group 3, SAH plus placebo; and Group 4, SAH plus rhEPO (each group consisted of eight rabbits). The rhEPO and placebo were administered to the rabbits after SAH had been induced, and S-100 protein levels in the CSF of these animals were measured at 24, 48, and 72 hours after the experimental procedure. In each group of animals levels of S-100 protein were compared with the mortality rate, neurological outcome, and neuronal ischemic damage. High S 100 protein levels were found in rabbits in Groups 2 and 3, which exhibited poor neurological status and harbored a high number of damaged cortical neurons. Favorable neurological outcome and significant reductions in total numbers of damaged neurons were observed in animals in Group 4 in which there were significantly lower S-100 protein concentrations compared with animals in Groups 2 and 3 (p < 0.001). CONCLUSIONS: The results of this study support the concept that determination of the S-100 protein level in CSF has prognostic value after SAH. The findings also confirm that rhEPO acts as a neuroprotective agent during experimental SAH. PMID- 11883843 TI - Total intravenous anesthesia for intraoperative monitoring of the motor pathways: an integral view combining clinical and experimental data. AB - OBJECT: Monitoring of descending corticospinal pathways by using motor evoked potentials (MEPs) has proven to be useful in preventing permanent neurological deficits during cranial and spinal procedures. Difficulties in interpretation of intraoperative changes in potentials may largely be attributed to the effects of anesthesia. Development of suitable intravenous anesthesia protocols specifically tailored for MEP monitoring, including plasma level target-controlled infusion (TCI), requires precise knowledge of the specific neurophysiological properties of the various agents. METHODS: The effects of alfentanil, sufentanil, fentanyl, remifentanil, thiopental, midazolam, etomidate, ketamine, and propofol on neurogenic and myogenic MEPs were evaluated in an integral study combining clinical data obtained in 40 patients and experimental investigations conducted in 140 animals. The dose-dependent modulation of MEPs after electrical and magnetoelectrical stimulation of the motor cortex was recorded from peripheral muscles and the spinal cord. The results were as follows: opioids, propofol, and thiopental suppressed myogenic, but not neurogenic MEPs in a dose-dependent fashion; remifentanil exerted the least suppressive effects. Etomidate and midazolam did not suppress myogenic MEP, even at plasma concentrations sufficient for anesthesia. Ketamine induced moderate reduction of compound muscle action potential amplitudes only at high doses. Remifentanil and propofol administered via TCI systems allowed recording of myogenic potentials within a defined target plasma concentration range. CONCLUSIONS: Development of standardized total intravenous anesthesia/TCI protocols by using anesthetic agents such as propofol, remifentanil, ketamine, and midazolam, which have favorable pharmacokinetic and neurophysiological properties, will enhance the quality of intraoperative MEPs and promote the use of MEP monitoring as a useful tool to reduce surgery-related morbidity. PMID- 11883844 TI - Apoptosis of T lymphocytes invading glioblastomas multiforme: a possible tumor defense mechanism. AB - OBJECT: The goal of this study was to investigate whether apoptosis occurs in T lymphocytes that invade Fas ligand (FasL)-expressing glioblastomas multiforme (GBMs) and if its induction could be mediated by Fas. METHODS: Apoptotic T lymphocytes were detected in GBMs by using detection of cell-type markers combined with active caspase-3 immunohistochemical analysis, a recently introduced apoptosis-specific in situ ligation assay, as well as by examining morphological criteria. Apoptotic T cells expressed Fas and were localized in the vicinity or in direct contact with FasL-expressing tumor cells. The T lymphocytes were undergoing apoptosis in spite of Bcl-2 expression. Expression of Bax was also detected in dying T cells, which can explain the absence of the protective effect of Bcl-2. because Bax inhibits Bcl-2 death-repressor activity. CONCLUSIONS: On the basis of the data presented in this paper, the authors suggest that GBM cells that express FasL can induce apoptosis in invading immune cells. This phenomenon may play an important role in these tumors' maintenance of immune privilege and evasion of immune attacks. Awareness of this phenomenon should be helpful for the development of novel strategies for treatment of malignant gliomas. PMID- 11883845 TI - Chronic granulomatous neuritis in idiopathic trigeminal sensory neuropathy. Report of two cases. AB - Idiopathic trigeminal sensory neuropathy is a clinically benign disorder in which the main feature is facial numbness limited to the territory of one or more divisions of the trigeminal nerve; the disorder persists for a few weeks to several years. and no underlying disease can be identified. Magnetic resonance (MR) imaging findings are occasionally consistent with a small trigeminal neuroma of the left gasserian ganglion associated with idiopathic trigeminal sensory neuropathy. The authors report on two patients who were treated using a skull base approach in which the gasserian ganglion was exposed and the lesion was removed. The pathological diagnosis was chronic granulomatous neuritis. The authors conclude that, in patients with MR findings suggestive of a small trigeminal neuroma, benign idiopathic trigeminal sensory neuropathy should also be considered in the differential diagnosis. A conservative approach featuring sequential MR imaging studies may avoid an unnecessary surgical exploration. PMID- 11883846 TI - Simultaneous intrastriatal and intranigral fetal dopaminergic grafts in patients with Parkinson disease: a pilot study. Report of three cases. AB - The main neural transplantation strategy in Parkinson disease (PD) has been focused on reinnervating the striatum. The clinical results reported in patients who receive transplants have been limited and do not justify the use of neural transplantation as a routine therapeutic procedure for PD. Identifying the optimal target for transplantation may be one of the critical factors for optimizing clinical outcomes. Evidence from preclinical studies indicates that simultaneous intrastriatal and intranigral grafts (double grafts) may produce a more complete functional recovery. The authors report the clinical and positron emission tomography (PET) scanning results in three patients enrolled in a safety and feasibility pilot study who received double grafts and who have been followed for up to 13 months posttransplantation. Patients included in the study had idiopathic PD. All patients underwent detailed assessments before and after surgery, in accordance with the Core Assessment Program for Intracerebral Transplantation. The patients received implants of fetal mesencephalic cell suspensions in the putamen and substantia nigra (SN) bilaterally. There were no intraoperative or perioperative complications. Follow-up PET scans demonstrated an increase in the mean fluorodopa uptake constant values in the putamen and SN 12 months postsurgery. Improvements were also noted in the total Unified Parkinson's Disease Rating Scale, Hoehn and Yahr, Schwab and England, and pronation/supination scores after transplantation. The authors demonstrate the feasibility of reinnervating the SN and striatum by using a double transplant strategy in humans. PMID- 11883847 TI - Bilateral cavernous sinus actinomycosis resulting in painful ophthalmoplegia. Case report. AB - The authors report the successful treatment of a 42-year-old man who suffered from recurrent painful ophthalmoplegia caused by bilateral cavernous sinus (CS) actinomycosis. A presumptive diagnosis of Tolosa-Hunt syndrome was made when he presented with left painful ophthalmoplegia. Recurrent ophthalmoplegia on the opposite side when steroid medications were tapered led to repeated imaging and a pterional craniotomy and biopsy sampling of the CS. These tests demonstrated acute inflammation and sulfur granules, which responded clinically and radiologically to parenterally administered penicillin therapy. Actinomycosis may present as a painful ophthalmoplegia with involvement of one or both CSs. Repeated imaging and possibly surgical exploration may be necessary to make a definitive diagnosis. PMID- 11883848 TI - Cranial root injury in glossopharyngeal neuralgia: electron microscopic observations. Case report. AB - Optical and electron microscopic examinations were made of a biopsy sample of the ninth and 10th cranial nerves obtained during posterior fossa surgery for the relief of pain in a patient suffering from glossopharyngeal neuralgia (GN). Pathological findings, which were restricted to a small fraction of fascicles in the nerves, included large patches of demyelinated axons in close membrane-to membrane apposition to one another and zones of less severe myelin damage (dysmyelination). These observations, in the light of similar morphological changes observed in biopsy samples excised from patients with trigeminal neuralgia, and new information on the pathophysiological characteristics of injured peripheral nerve axons, can account for much of the symptomatology of GN. PMID- 11883849 TI - Cerebellar mutism associated with a midbrain cavernous malformation. Case report and review of the literature. AB - The authors report a case of cerebellar mutism arising from a hemorrhagic midbrain cavernous malformation in a 14-year-old boy. No cerebellar lesion was identified; however, edema of the dorsal midbrain was noted on postoperative magnetic resonance images. Dysarthric speech spontaneously returned and then completely resolved to normal speech. This case provides further evidence for the theory that involvement of the dentatothalamic tracts, and not a cerebellar lesion per se, is the underlying cause of "cerebellar" mutism. PMID- 11883851 TI - Bilateral thalamic deep brain stimulation for the treatment of head tremor. Report of two cases. AB - Isolated head tremor is rare, but can be disabling. The authors' experience with the treatment of limb tremor due to essential tremor led them to consider using bilateral thalamic deep brain stimulation (DBS) in two patients presenting only with disabling head tremor. One patient exhibited no peripheral tremor and the other displayed only a slight upper-limb tremor. Both patients underwent placement of units that apply simultaneous bilateral thalamic DBS. Surgical targets were verified by using intraoperative macrostimulation, and the stimulators were implanted during the same surgery. Patients were videotaped preoperatively and at 2, 4, 6, and 9 months postoperatively during periods in which the stimulators were turned on and off. Videotapes were randomized and rated for resting, postural, and action tremors according to the Fahn clinical rating scale for tremor. Because this scale is not designed for head tremor, the patients were also evaluated on the basis of a functional scale that reflected their quality of life and the amount of disability caused by head tremor. Both patients experienced no tremor after their stimulators were turned on and properly adjusted at the 6th postoperative week. The patients were followed for a total of 9 months and results remained stable throughout this period. No complications were encountered. Bilateral thalamic DBS appears to be an effective and safe treatment for isolated head tremor in patients with essential tremor. The authors present a scale for the functional assessment of head tremor. PMID- 11883850 TI - Primary low-grade B-cell lymphoma of mucosa-associated lymphoid tissue of the dura mimicking the presentation of an acute subdural hematoma. Case report and review of the literature. AB - The authors present the case of a 64-year-old woman who experienced a left hemiparesis. An initial diagnosis of subdural hematoma was made based on results of computerized tomography scanning. Subsequent magnetic resonance imaging indicated an extraaxial meningioma. Histological findings confirmed an extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT). The authors outline the natural history of central nervous system lymphomas and of MALT lymphomas in other tissues. They review seven previously reported cases and emphasize the importance of recognizing these tumors as a distinct clinicopathological entity. PMID- 11883852 TI - Intracranial meningeal melanocytoma associated with ipsilateral nevus of Ota. Case report. AB - In this report, the authors review the case of a man with a neurocutaneous syndrome. He presented with an intracerebral melanocytoma associated with a blue nevus of the scalp; its location and its appearance during childhood supported the diagnosis of a nevus of Ota. Meningeal melanocytomas are increasingly being diagnosed, but remain rare. Primary meningeal malignant melanoma is the first differential diagnosis to eliminate. Despite their common embryonic origin. the association of a melanocytoma with a nevus of Ota is rare. A nevus of Ota exhibits the same melanocytic proliferation and affects the trigeminal nerve territory. An ocular effect is not always observed. In contrast to an ocular lesion, a nevus of Ota rarely transforms into a malignant melanoma. It is found only among caucasians. During 4 years of follow-up review after surgery, the patient remained asymptomatic. Other than antiepileptic therapy, he received no complementary treatment and cerebral imaging revealed no evidence of recurrence. PMID- 11883853 TI - Endovascular treatment of an internal carotid artery pseudoaneurysm following transsphenoidal surgery. Case report. AB - Internal carotid artery (ICA) pseudoaneurysm formation following transsphenoidal surgery is a rare but potentially lethal complication. Direct surgical repair with preservation of the ICA may be difficult. The feasibility of endovascular coil embolization with parent artery preservation for an iatrogenic ICA pseudoaneurysm is undefined. A 40-year-old man was referred to the authors' institution after identification of a pseudoaneurysm of the left ICA following transsphenoidal resection of a pituitary macroadenoma. The pseudoaneurysm was treated via an endovascular approach that included stent-assisted coil embolization of the lesion. Follow-up angiographic studies obtained 1 year later demonstrated complete occlusion of the aneurysm, and the patient remains asymptomatic. Stent-assisted coil embolization of this iatrogenic pseudoaneurysm was successful in achieving complete, angiographically confirmed aneurysm obliteration, with preservation of the ICA and short-term prevention of hemorrhage or carotidcavernous fistula. The endovascular method provided an effective, relatively low-risk treatment for this difficult lesion, and was an excellent alternative to direct surgical repair. Nonetheless, long-term follow-up review is required before definitive treatment recommendations can be made. PMID- 11883854 TI - A new method of ultrasonic guidance of neuroendoscopic procedures. Technical note. AB - The authors present a newly designed device for ultrasonic guidance of neuroendoscopic procedures. It consists of a puncture adapter that attaches to a rigid endoscope having an outer diameter of 6 mm and is mounted on a small, bayonet-shaped ultrasound probe. This adapter directs the movement of the endoscope precisely within the ultrasonic field of view. The targeted region is identified by transdural insonation via an enlarged single burr-hole approach, and the endoscope is tracked in real time throughout its approach to the target. The procedure has been performed in 10 patients: endoscopic ventriculocystostomy in four cases; removal of a colloid cyst of the third ventricle in two cases; and intraventricular tumor biopsy, intraventricular tumor resection, third ventriculostomy, and removal of an intraventricular hematoma in one case each. The endoscope was depicted on ultrasonograms as a hyperechoic line without disturbing echoes and, consequently, the target (cyst, ventricle, or tumor) was safely identified in all but one case, in which intraventricular air hid a colloid cyst in the foramen of Monro. The method presented by the authors proved to be very effective in the guidance and control of neuroendoscopic procedures. Combining this method with image guidance is recommended to define the entry point of the endoscope precisely. PMID- 11883855 TI - Craniofacial surgery. PMID- 11883856 TI - Posterior fossa aneurysms. PMID- 11883857 TI - Posterior fossa aneurysms. PMID- 11883858 TI - Cavernous sinus meningioma. PMID- 11883859 TI - Parkinson disease. PMID- 11883860 TI - The puzzle of hormone replacement therapy (HRT) and cardiovascular disease (CVD). PMID- 11883861 TI - Soluble CD4 concentrations predict relapse of post-partum thyroiditis. AB - Post-partum thyroiditis (PPT) is a common autoimmune thyroid disorder which results in significant morbidity at a critical time of a woman's life. The presence of anti-thyroglobulin (anti-TG) and, more so, anti-thryroperoxidase (anti-TPO) antibodies in the first trimester of pregnancy has been reported to forecast subsequent PPT. Despite their predictive value, these tests lack in specificity. We have sought to find an alternative that is more specific and, ideally, which could be tested immediately proximate to the event. We have taken advantage of the high recurrence rate of PPT in subsequent pregnancies to perform a prospective study of serum soluble CD4 (sCD4) and CD8 (sCD8) levels in 22 pregnant women who had at least one previous episode of PPT. This group was matched with 21 pregnant women of comparable age with no evidence of thyroid disease. Both groups of women were sampled in each of the three trimesters of pregnancy, 1 month, 3 months and 6 months post-partum for sCD4, sCD8, thyroid function parameters and antibodies. Twelve of the 22 women with previous PPT had recurrent disease; they were more likely to be cigarette smokers and to have a family history of autoimmune disorders (p<0.05, for both) than those who did not. Half of these women had high anti-TG or anti-TPO each in the first trimester compared to none among those without recurrent PPT and 2/21 controls. Serum sCD8 levels showed no changes over the observation points among the two PPT patient subsets and were comparable to those of the controls. By contrast, serum sCD4 concentrations showed divergent changes in the group with recurrent PPT in the course of pregnancy and postpartum period compared to those without disease recurrence and controls: sCD4 failed to show the physiological fall in the third trimester of pregnancy [19.0+/-1.7 (+/-SD) U/ml vs 15.6+/-2.3 U/ml in controls, NS]. This trend was continued into the first month post-partum when sCD4 levels were clearly higher than in controls (22.1+/-2.6 U/ml compared to 17.9+/-1.9 U/ml in controls, p<0.001) and well before the episode of PPT. An sCD4 serum level outside the 95% reference range at 1 month post-partum (9/12 in recurrent PPT, 1/21 in controls) yields a relative risk of 6.9 (chi2=14.67, p<0.001) compared to 3.3 for first trimester thyroid antibody positivity (p=0.029). In summary, we describe a reliable test for forecasting PPT that can be obtained immediately proximate to the possible event. If our findings are verified in larger studies, the measurement of serum sCD4 concentration drawn in the first month post-partum may prove an ideal test for population screening for impending PPT. PMID- 11883862 TI - Differentiated thyroid cancer in children and adolescents. AB - In this retrospective study we analyzed cancer characteristics and outcome in a consecutive series of 48 young patients (< or =20 yr of age) with a differentiated thyroid cancer (DTC), observed during the period 1977-1998. In none of them was thyroid cancer related to ionizing radiation. The median age was 18.1 yr, range 7-20, and the female/male ratio was 2.5/1. Papillary thyroid cancer (PTC) occurred in 83% and follicular thyroid cancer (FTC) in 17% of cases. All patients underwent total or near total thyroidectomy plus pre- and/or paratracheal lymphnode dissection. Surgery complication rate was low (4% permanent hypoparathyroidism; no permanent lesion of recurrent laryngeal nerve). Extrathyroid disease was present in 52% of patients with PTC and in 50% of patients with FTC, while nodal metastases were present in 62.5% of patients with PTC and in 12.5% of patients with FTC. Lung metastases occurred in 10 patients with PTC (25%) and in none with FTC. Twenty-one patients required radioiodine treatment for metastatic disease: 11 patients for relapsing lymph-node metastases, 4 patients for lung metastases, 6 patients for both lymph-node and lung metastases. After a mean follow-up of 85+/-12 months all patients followed regularly (no.=47) were alive; 37 patients (79%) were free of disease and 10 (21%) had residual disease. Our results indicate that non-radiation-related DTC occurring in young patients often presents at an advanced stage. For this reason, although the prognosis is usually good in these patients, we believe that total or near total thyroidectomy with lymphadenectomy is always the required initial surgical treatment. PMID- 11883863 TI - The finding of a somaticdeletion in RET exon 15 clarified the sporadic nature of amedullary thyroid carcinoma suspected to be familial. AB - Medullary thyroid carcinoma (MTC) occurs both sporadically and in the autosomal dominantly inherited multiple endocrine neoplasia (MEN) type 2 syndromes. The distinction between both is important for future clinical management. We report a family initially described as a familial MTC by pentagastrin stimulation test and clinical outcome, in which we found a 12 bp deletion within the catalytic domain of the protooncogene RET in the index case tumor alone. Linkage study suggests that it is a sporadic MTC. Therefore, in view of these results, in kindred with just one MTC case, borderline pentagastrin test values must be carefully assessed. In addition, this and other mutations can help us to understand some features about domains that play an important role in the normal function of this tyrosine kinase receptor and involved in MTC. PMID- 11883864 TI - Thyroxine binding to members and non-members of the serine protease inhibitor family. AB - Partition of T4 to plasma proteins is classically attributed only to TBGC approximately/= 70%), transthyretin (approximately/= 15%) and albumin (approximately/= 10%), based on zone electrophoresis. Since TBG migration spans the alpha1 and alpha2 regions, and since HDL, which have alpha1 migration, transport approximately/= 4% of circulating T4, other alpha-globulins could bind T4 and "contaminate" the TBG area. Hence, we determined the association of [125I]T4 to TBG and, for comparison, to transthyretin and albumin. Sera from 50 normolipidemic individuals were equilibrated with [125I] T4 and analyzed by both zone electrophoresis and radioimmunoprecipitation with specific antisera. Transthyretin-T4 or albumin-T4 bindings as assessed by the two methods agreed, while TBG-T4 did not, because other alpha-globulins carrying T4 with low affinity co-migrated with TBG. Some, but not all, of these alpha-globulins belong to the same superfamily of TBG and also bind steroid hormones. PMID- 11883865 TI - Radioiodine ablation and therapy in differentiated thyroid cancer under stimulation with recombinant human thyroid-stimulating hormone. AB - We investigated whether recombinant human TSH (rhTSH) safely and effectively induces uptake of high-dose 131-iodine (131I) to ablate thyroid remnant or treat disease, in patients with well-differentiated thyroid carcinoma. Eleven consecutive patients unable to tolerate thyroid hormone withdrawal received one im injection of 0.9 mg rhTSH on 2 consecutive days before receiving 4000 MBq (approximately 108 mCi) radioiodine orally. Eight patients received one, and 3 patients 2 courses. Our series comprised 7 women and 4 men (mean age, 78 yr, range: 56-87 yr). Ten patients had undergone total or near-total thyroidectomy up to 19 yr earlier. rhTSH-stimulated single course radioiodine with the intention to ablate thyroid remnant was performed in 3 patients, with following estimation of radioiodine uptake and TG measurements. Of another 8 patients given this treatment palliatively, 5 had radiological, clinical and/or laboratory response, including: 80% decreased pathological uptake between treatment courses; pronounced decrease in bone pain; diminished symptoms; improved physical condition and quality of life; lower serum TG concentration; and/or normalization of TG recovery test. Two patients with small lung metastases on computed tomography had no detectable radioiodine uptake or other response; they also lacked uptake after withdrawal-stimulated radioiodine treatment. Despite being elderly and frail, patients generally tolerated treatment well; rhTSH caused nausea in one patient and transiently increased pain in bone and soft tissue lesions in another. We conclude that rhTSH-stimulated high-dose radioiodine for remnant ablation or tumor treatment is safe, feasible and seemingly effective, enhancing quality of life and offering reasonable palliation in patients with advanced disease. PMID- 11883866 TI - Unusual clinical manifestation of pheochromocytoma in a MEN2A patient. AB - A case of unusual clinical manifestation of pheochromocytoma in a type 2A multiple endocrine neoplasia (MEN2A) patient is presented. A 27-year-old man affected by MEN2A syndrome, complaining of anxiety and depression, was admitted in our Division. Past medical history included a total thyroidectomy for medullary carcinoma in 1985, and left adrenalectomy for pheochromocytoma in 1994. Blood pressure was 130/ 85 mmHg without orthostatic hypotension and pulse rate was 72 beats/min. Laboratory data revealed thyroid hormones and carcinoembryonic antigen (CEA) in the normal range and high basal serum calcitonin levels (158 pg/ml). Plasma catecholamines and vanillylmandelic acid resulted in normal levels but epinephrine/norepinephrine ratio was elevated (0.65). The glucagon stimulation test showed positive clinical and biochemical response. Magnetic resonance imaging (MRI) and meta-iodobenzylguanidine (MIBG) scintiscan confirmed the presence of bilateral adrenal masses. Bilateral adrenalectomy by laparoscopic anterior approach was performed. Histology was consistent with adrenal pheochromocytomas. After surgical approach, psychiatric findings disappeared and did not recur at follow-up in spite of no medication for two years. In conclusion, bilateral pheochromocytoma is more frequent in MEN2A syndrome and probably understimated if the follow-up is not prolonged. In these cases clinical features are often aspecific and basal hormonal data may be normal in a great number of patients. Therefore long-term observation is justified in these patients. Pheochromocytoma was described as the "great mimic" for the numerous subjective manifestations. Differential diagnosis among typical features of neuropsychiatric disorders and pheochromocytoma must be considered. PMID- 11883867 TI - Abnormalities of GH secretion in a young girl with Floating-Harbor syndrome. AB - We present a 9.1-year-old girl of Calabrian (Italy) ancestry, with clinical features (cranio-facial dysmorphism, short stature with delayed bone age and speech delay) suggesting the diagnosis of Floating-Harbor syndrome (FHS). Physical examination showed: height 113.9 cm (-2.9 SD), with a parent's target of 156.2 cm (+1.0 SD), weight 20.7 kg, BMI 16.0 (-0.04 SD), and many phenotypic abnormalities: long eyelashes, large bulbous nose with broad nasal bridge, short philtrum, moderately broad mouth, tooth folding and malocclusion, posteriorly rotated ears, low posterior hair line, short neck, clinodactyly of the 5th finger and hyperextensible finger joints. Diffused hyperpigmentation and hypertrichosis with sporadic pubic terminal hairs, but neither clitoromegaly nor other signs of hyperandrogenism and/or precocious puberty, were observed (T1, P1). Carpal bone evaluation showed a delayed bone age (TW2: 5-5/10, - 3.6 yr) and the statural age/bone age ratio was 1.1. Other dysmorphic syndromes were excluded on the basis of clinical evidence, also evaluated by a computer-assisted search (P.O.S.S.U.M. version 3.5, 1992). Analysis of chromosome 22 by the FISH method, using specific probes Cos29 and Tuple1, excluded microdeletions in the region 22q11.2, typical of Velo-cardio-facial syndrome. In this case, we report the impairment of serum GH responsiveness (GH baseline values: 0.2-1.9 ng/ml) to the administration of oral 150 microg clonidine [peak 4.7 ng/ml, normal values (nv)>10 ng/ml] and oral 4 mg dexamethasone (8.1 ng/ml, nv>10 ng/ml). Moreover, the evaluation of spontaneous 24-h GH secretion (Carmeda AB, Stockholm, Sweden) showed low mean GH levels (1.75 ng/ml, nv>3.0 ng/ml), with a maximum sleep-related peak of 2.8 ng/ml. Serum IGF-1 values were in the low-normal range (80-176 ng/ml, nv 133-626 ng/ml). While in FHS the cranio-facial features minimize with advancement of age, the impairment of growth velocity is permanent and results in severe dwarfism. In our case, treatment with recombinant GH (0.10 U/kg/day), administered by a needle free device, induced a dramatic increase of growth velocity, increasing the height from -2.8 to -1.9 SD after 18 months, thus indirectly confirming a role of GH deficiency in the pathogenesis of FHS dwarfism. PMID- 11883868 TI - Pituitary carcinoma: report of an exceptional case and review of the literature. AB - Pituitary carcinomas are exceptional tumors and constitute 0.1 to 0.2% of pituitary tumors. Their definition includes well-established criteria but distant metastasis is the hallmark required for diagnosis. We report the fourth case of gonadotropic pituitary carcinoma described in the literature. This case illustrates the dramatic outcome of these tumors. The most interesting feature of our case was the loss of differentiation with time, established by retrospective analysis of the primary tumor surgically treated 15 years earlier. Most of the previously reported cases exhibited a majority of adrenocoticotropin and non functioning pituitary tumors. However, the frequency of non-functioning tumors seems smaller than previously believed. In the discussion, we stress the need to detect these very aggressive tumors as early as possible and identify treatments to improve the dramatic course of these carcinomas. PMID- 11883871 TI - Pre-natal treatment of congenital adrenal hyperplasia and fetal malformations. PMID- 11883869 TI - Dementia: a neuroendocrine perspective. AB - The etiology of Alzheimer's disease (AD) has not been as yet completely defined. Genetic, environmental and neurophysiological aspects should all be taken into account. The disease has also neuroendocrine implications, some of which are discussed in this review. It is known that stress and glucocorticoids may affect neurone survival. On the contrary, some data indicate that DHEA and DHEAS exert a neuroprotective action. In AD, changes in hypothalamic-pituitary-adrenal axis function have been reported. Experimental and clinical evidence indicates that glucocorticoid hypersecretion and DHEAS levels decrement may add to hippocampal dysfunction in aging and in AD. Glucocorticoid and beta-amyloid concur in the mechanism of neurone damage, as well as excitatory amino acids (EAA), Ca++ and reactive oxygen species (ROS). The neuroprotective effects exerted by IGFs are also hindered in aging and even more in AD. Production and biological actions of IGFs are negatively influenced by cortisol hypersecretion and DHEAS decrease in patients with AD. PMID- 11883870 TI - Comparison between buserelin and dexamethasone testing in the assessment of hirsutism. AB - Many hirsute women may present a form of functional ovarian hyperandrogenism (FOH), since they show an exaggerated 17-hydroxyprogesterone (17-OHP) response to GnRH agonists administration. As the failure of dexamethasone to reduce testosterone levels may be indicative of an ovarian source of androgen secretion, we evaluated the usefulness of dexamethasone suppression test, in comparison with buserelin challenge, in the assessment of hirsutism. Twenty-seven hirsute women (aged 15-42 yr) underwent ACTH and buserelin tests: 4 patients were heterozygotes for 21-OH deficiency and 8 patients were affected with FOH: 2 of the patients with hyperresponse to buserelin also had 21-hydroxylase deficiency. The results of the dexamethasone suppression test (2 mg/day for 7 days) were compared to those obtained after buserelin test. Basal T and delta4 levels (mean+/-SE) were higher than in controls (4.2+/-0.5 vs 2.2+/-0.2 nmol/l and 10.9+/-0.9 vs 5.9+/ 0.6 nmol/l, p<0.02), while no differences were found in 17-OHP and DHEAS levels. A significant reduction (p<0.001) in T (1.8+/-0.4 nmol/l), delta4 (3.2+/-0.5 nmol/l) and DHEAS levels (2.4+/-0.3 micromol/l) was observed at the 3rd day of dexamethasone administration and no differences between sampling at 3rd, 5th and 7th day were found. Serum T was not suppressed in 6 cases, delta4 and DHEAS levels in 3 and 1 of them, respectively. Buserelin injection caused an excessive 17-OHP response in 8 patients, only 4 of them did not reduce T levels during dexamethasone. The sensitivity and specificity of the dexamethasone suppression test, with respect to the buserelin test, were 50% and 89%, respectively. In conclusion, 37% of hirsute patients had an abnormal responsiveness to buserelin and/or ACTH tests, indicating that hormonal investigations are mandatory. An ovarian origin of hirsutism was identified by buserelin test in 30% of patients and by dexamethasone in 22% of cases; only 4 of 8 patients showed concordant results to both tests. Therefore, buserelin challenge seems a more useful, cost effective and less time consuming tool than dexamethasone administration in order to recognize the possible ovarian origin of hyperandrogenism. PMID- 11883872 TI - Lymphocytic hypophysitis and diabetes insipidus in non-pregnant women. PMID- 11883873 TI - A novel approach for the analysis of DAZ gene copy number in severely idiopathic infertile men. AB - The deleted-in-azoospermia (DAZ) gene family constitutes the major candidate for the AZFc (azoospermia factor c) phenotype of male infertility, being deleted in about 10% of azoospermic and severely oligozoospermic subjects. Four DAZ genes are arranged in two clusters in AZFc, and standard analysis by PCR cannot distinguish among the different copies. Therefore only deletions of the entire gene cluster can be identified. We developed a PCR amplification-restriction digestion assay able to distinguish from DAZ genes for single nucleotide variants. Then we applied this approach to screen a group of idiopathic infertile men in which the DAZ genes presence was previously assessed by standard PCR analysis. Two patients out of 25 showed deletion of two copies of DAZ (DAZI and 2), suggesting that this mutation was actually the cause of spermatogenic damage. This preliminary screening demonstrates that deletions of copies of DAZ genes may be often found in severely infertile men and it strengthens the role of this gene family in spermatogenesis. Furthermore, this simple method, being able to distinguish among the different DAZ copies, could be used to screen a larger number of patients and to perform a more accurate diagnosis. PMID- 11883874 TI - Population genetic structure of the malaria mosquito Anopheles arabiensis across Nigeria suggests range expansion. AB - Ten microsatellite loci, four located within and six outside chromosome inversions, were employed to study the genetic structure of Anopheles arabiensis across the ecological zones of Nigeria (arid savannah in the north gradually turns into humid forest in the south). Regardless of location within or outside inversions, genetic variability at all loci was characterized by a reduction in both the number of alleles per locus and heterozygosity from savannah to forest. Across all loci, all but one allele in the forest also occurred in the savannah, whereas at least 78 alleles in the savannah were missing in the forest. Genetic differentiation increased with geographical distance; consequently, genetic distances between zones exceeded those within zones. The largest genetic distances were between localities at the extremes of the transect (range F(ST) = 0.196-0.258 and R(ST) = 0.183-0.468) and were as large as those between A. arabiensis and Anopheles gambiae s.s. Gene flow across the country was very low, so that Nm between the extremes of the transect was < 1. These data suggest that A. arabiensis has extended its range from the savannah into the forest during which it experienced a reduction in effective population size due to sequential founder effects. Gene flow post range expansion appears too restricted by geographical distance to homogenize the gene pool of A. arabiensis across Nigeria. PMID- 11883875 TI - Random amplified polymorphic DNA and amplified fragment length polymorphism assessment of genetic variation in Nicaraguan populations of Pinus oocarpa. AB - Pinus oocarpa is the most widely distributed pine species of Mexico and Central America. The natural populations of Nicaragua have been affected by extensive human activities. As a consequence, their size has been reduced, and there is a serious threat to the development of mature woodland. Knowledge of population structures and the genetic diversity of the species is required for the design of sustainable use and conservation strategies. Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers were used to assess the genetic variation among 10 populations from three geographical regions of Nicaragua. Both markers revealed high levels of diversity in these populations. G(ST) values and analyses of molecular variance (AMOVA) found that most variation was within populations but there is still a significant differentiation between populations indicating that the populations sampled cannot be considered a single panmictic unit. The partitions created by AMOVA also showed that there was little differentiation between populations of different regions, although cluster analyses based on RAPDs and AFLPs indicated a closer relationship among most of the populations from a same geographical region. Management of P. oocarpa in Nicaragua should be aimed to maintain the high degree of genetic variation within individual populations that is still observed even in some of these highly degraded populations. PMID- 11883876 TI - Genetic divergence of peripherally disjunct populations of the gastropod Batillariella estuarina in the Houtman Abrolhos Islands, Western Australia. AB - Geographically disjunct populations are unusual in marine species, but the Houtman Abrolhos Islands, Western Australia, provide opportunities to study highly disjunct peripheral isolates of several species. The intertidal snail Batillariella estuarina occurs in isolated tidal ponds in the Abrolhos Islands, where it is at its northern limit, disjunct from mainland populations by 600-900 km. The species is thus disjunct both geographically and among the peripherally isolated populations in the Abrolhos Islands. Comparisons of allozymes at 11 polymorphic loci were made among populations from 10 ponds in the Abrolhos Islands and six sites from relatively continuous tidal flats at Albany, 900 km away, the nearest major set of populations. Among all 16 populations, subdivision was high (FST = 0.455). Although there were subtle differences between the geographical regions, the large majority of divergence occurred among the isolated ponds in the Abrolhos (FST = 0.441), and divergence on the tidal flats at Albany was only moderate (FST = 0.085). Characteristic of peripheral isolates, the pond populations have less polymorphism and fewer alleles than the more connected populations at Albany. Combined with evidence of genetic divergence in the gastropods Bembicium vittatum and Austrocochlea constricta, which have very similar geographical distributions to that of B. estuarina, these results indicate the potential evolutionary significance of peripherally isolated marine populations in the unusual habitats of the Abrolhos Islands. PMID- 11883877 TI - Phylogenetics, genome diversity and origin of modern leopard, Panthera pardus. AB - Leopards, Panthera pardus, are widely distributed across southern Asia and sub Saharan Africa. The extent and phylogeographic patterns of molecular genetic diversity were addressed in a survey of 77 leopards from known geographical locales representing 13 of the 27 classical trinomial subspecies. Phylogenetic analysis of mitochondrial DNA sequences (727 bp of NADH5 and control region) and 25 polymorphic microsatellite loci revealed abundant diversity that could be partitioned into a minimum of nine discrete populations, tentatively named here as revised subspecies: P. pardus pardus, P. p. nimr, P. p. saxicolor, P. p. fusca, P. p. kotiya, P. p. delacouri, P. p. japonensis, P. p. orientalis and P. p. melas. However, because of limited sampling of African populations, this may be an underestimate of modern phylogeographic population structure. Combined phylogeographic and population diversity estimates support an origin for modern leopard lineages 470,000-825,000 years ago in Africa followed by their migration into and across Asia more recently (170,000-300,000 years ago). Recent demographic reductions likely have led to genetic impoverishment in P. p. orientalis and in the island subspecies P. p. kotiya. PMID- 11883878 TI - High within-population mitochondrial DNA variation due to microvicariance and population mixing in the land snail Euhadra quaesita (Pulmonata: Bradybaenidae). AB - A high level of geographical variation at an exceptionally fine scale was detected in the mitochondrial 16S ribosomal RNA genes of a land snail species Euhadra quaesita from the Kanto region of Japan. In total, 50 haplotypes were detected from 27 populations, with most sample sites possessing private alleles. In some individual populations the different haplotypes do not fall as a monophyletic group, so that some of the haplotypes are phylogenetically distant, differing from each other by > 10%. In contrast, phylogenetically similar haplotypes were found in separate sites at long distances from their main distribution. Together, these findings strongly suggest that contraction and expansion of populations has occurred repeatedly in the past. The subsequent expansion of populations and migration from different areas may have mixed distant populations. This repetition of isolation and mixing has resulted in an exceptionally fine scale of geographical variation, and the accumulation of high genetic diversity within and between populations of this species. PMID- 11883879 TI - Global relationships amongst black-browed and grey-headed albatrosses: analysis of population structure using mitochondrial DNA and microsatellites. AB - The population structure of black-browed (Thalassarche melanophris and T. impavida) and grey-headed (T. chrysostoma) albatrosses was examined using both mitochondrial DNA (mtDNA) and microsatellite analyses. mtDNA sequences from 73 black-browed and 50 grey-headed albatrosses were obtained from five island groups in the Southern Ocean. High levels of sequence divergence were found in both taxa (0.55-7.20% in black-browed albatrosses and 2.10-3.90% in grey-headed albatrosses). Black-browed albatrosses form three distinct groups: Falklands, Diego Ramirez/South Georgia/Kerguelen, and Campbell Island (T. impavida). T. melanophris from Campbell Island contain birds from each of the three groups, indicating high levels of mixture and hybridization. In contrast, grey-headed albatrosses form one globally panmictic population. Microsatellite analyses on a larger number of samples using seven highly variable markers found similar population structure to the mtDNA analyses in both black-browed and grey-headed albatrosses. Differences in population structure between these two very similar and closely related species could be the result of differences in foraging and dispersal patterns. Breeding black-browed albatrosses forage mainly over continental shelves and migrate to similar areas when not breeding. Grey-headed albatrosses forage mainly at frontal systems, travelling widely across oceanic habitats outside the breeding season. Genetic analyses support the current classification of T. impavida as being distinct from T. melanophris, but would also suggest splitting T. melanophris into two groups: Falkland Islands, and Diego Ramirez/South Georgia/Kerguelen. PMID- 11883880 TI - A molecular phylogenetic analysis of diversification in Amazonian Anolis lizards. AB - We present a mitochondrial DNA (mtDNA) haplotype phylogeny for Amazonian Anolis lizards, including geographical sampling within four species distributed across the Amazon basin (A. fuscoauratus, A. nitens, A. ortonii and A. punctatus). Approximately 1500 bp of mtDNA encoding ND2, COI and four transfer RNAs (tRNAs) are reported for 39 specimens representing four to five populations of each widespread species, plus eight outgroups. These new sequences are aligned with eight previously published sequences, yielding 914 variable characters and 780 parsimony-informative characters. Phylogenetic analyses using maximum parsimony and maximum likelihood reject the hypothesis that Amazonian anoles form a monophyletic group excluding Central American and Caribbean anoles, and suggest multiple faunal exchanges among these regions. Haplotype divergence among geographical populations of A. nitens, whose variation was influential in formulating the Pleistocene refuge hypothesis of Amazonian speciation, is very large (13-22% sequence difference), suggesting that these populations separated well before the Pleistocene. Haplotype divergences among geographical populations of A. fuscoauratus (3-4%), A. punctatus (4-9%) and A. ortonii (6-8%) also indicate pre-Pleistocene differentiation within each species, but temporally incongruent patterns among species. PMID- 11883881 TI - Organelle DNA phylogeography of Cycas taitungensis, a relict species in Taiwan. AB - The phylogegraphic pattern of Cycas taitungensis, an endemic species with two remaining populations in Taiwan, was investigated based on genetic variability and phylogeny of the atpB-rbcL noncoding spacer of chloroplast DNA (cpDNA) and the ribosomal DNA (rDNA) internal transcribed spacer (ITS) of mitochondrial DNA (mtDNA). High levels of genetic variation at both organelle loci, due to frequent intramolecular recombination, and low levels of genetic differentiation were detected in the relict gymnosperm. The apportionment of genetic variation within and between populations agreed with a migrant-pool model, which describes a migratory pattern with colonists recruited from a random sample of earlier existing populations. Phylogenies obtained from cpDNA and mtDNA were discordant according to neighbour-joining analyses. In total four chlorotypes (clades I-IV) and five mitotypes (clades A-E) were identified based on minimum spanning networks of each locus. Significant linkage disequilibrium in mitotype-chlorotype associations excluded the possibility of the recurrent homoplasious mutations as the major force causing phylogenetic inconsistency. The most abundant chlorotype I was associated with all mitotypes and the most abundant mitotype C with all chlorotypes; no combinations of rare mitotypes with rare chlorotypes were found. According to nested clade analyses, such nonrandom associations may be ascribed to relative ages among alleles associated with the geological history through which cycads evolved. Nested in networks as interior nodes coupled with wide geographical distribution, the most dominant cytotypes of CI and EI may represent ancestral haplotypes of C. taitungensis with a possible long existence prior to the Pleistocene glacial maximum. In contrast, rare chlorotypes and mitotypes with restricted and patchy distribution may have relatively recent origins. Newly evolved genetic elements of mtDNA, with a low frequency, were likely to be associated with the dominant chlorotype, and vice versa, resulting in the nonrandom mitotype-chlorotype associations. Paraphyly of CI and EI cytotypes, leading to the low level of genetic differentiation between cycad populations, indicated a short period for isolation, which allowed low possibilities of the attainment of coalescence at polymorphic ancestral alleles. PMID- 11883882 TI - Phylogeography of the red-tailed chipmunk (Tamias ruficaudus), a northern Rocky Mountain endemic. AB - The northern Rocky Mountains have experienced a complex history of geological events and environmental fluctuation, including Pleistocene glaciation. To provide an initial assessment of the genetic impact of this history on the regional biota we estimated phylogenetic relationships within Tamias ruficaudus, a regional endemic, from cytochrome b sequence variation using parsimony, maximum likelihood, and nested clade analysis. Analyses of sequence variation in 187 individuals from 43 localities across the distribution of T. ruficaudus indicate a history of vicariance events and range fluctuation consistent with successive periods of extensive Pleistocene glaciation in the northern Rocky Mountains. Intraspecific divergence levels (c. 4.7% uncorrected) and phylogenetic structure are consistent with a genealogical vicariance initiated prior to the Late Pleistocene, whereas nested clade analyses indicate more recent population history structured by both fragmentation and range expansion. A comparison of sequence variation with bacular morphology indicates that the two genetically and morphologically differentiated entities exhibit a zone of differential character introgression. Sequence data support a multiple refugia hypothesis and provide a phylogeographical case study for the ongoing synthesis of regional biogeography for northern Rocky Mountain endemics. PMID- 11883883 TI - Phylogeography of Kandelia candel in East Asiatic mangroves based on nucleotide variation of chloroplast and mitochondrial DNAs. AB - Vivipary with precocious seedlings in mangrove plants was thought to be a hindrance to long-range dispersal. To examine the extent of seedling dispersal across oceans, we investigated the phylogeny and genetic structure among East Asiatic populations of Kandelia candel based on organelle DNAs. In total, three, 28 and seven haplotypes of the chloroplast DNA (cpDNA) atpB-rbcL spacer, cpDNA trnL-trnF spacer, and mitochondrial DNA (mtDNA) internal transcribed spacer (ITS) were identified, respectively, from 202 individuals. Three data sets suggested consistent phylogenies recovering two differentiated lineages corresponding to geographical regions, i.e. northern South-China-Sea + East-China-Sea region and southern South-China-Sea region (Sarawak). Phylogenetically, the Sarawak population was closely related to the Ranong population of western Peninsula Malaysia instead of other South-China-Sea populations, indicating its possible origin from the Indian Ocean Rim. No geographical subdivision was detected within the northern geographical region. An analysis of molecular variance (AMOVA) revealed low levels of genetic differentiation between and within mainland and island populations (phiCT = 0.015, phiSC = 0.037), indicating conspicuous long distance seedling dispersal across oceans. Significant linkage disequilibrium excluded the possibility of recurrent homoplasious mutations as the major force causing phylogenetic discrepancy between mtDNA and the trnL-trnF spacer within the northern region. Instead, relative ages of alleles contributed to non-random chlorotype-mitotype associations and tree inconsistency. Widespread distribution and random associations (chi2 = 0.822, P = 0.189) of eight hypothetical ancestral cytotypes indicated the panmixis of populations of the northern geographical region as a whole. In contrast, rare and recently evolved alleles were restricted to marginal populations, revealing some preferential directional migration. PMID- 11883884 TI - A genetic analogue of 'mark-recapture' methods for estimating population size: an approach based on molecular parentage assessments. AB - Molecular polymorphisms have been used in a variety of ways to estimate both effective and local census population sizes in nature. A related approach for estimating the current size of a breeding population, explored here for the first time, is the use of genetic 'marks' reconstructed for otherwise unknown parents in paternity or maternity analyses of progeny arrays. This method provides interesting similarities and contrasts to traditional mark-recapture methods based on physical tags. To illustrate, this genetic method is applied to a population of painted turtles on the Mississippi River to estimate the number of successfully breeding males. Non-genetic mark-recapture approaches were also applied to animals trapped at this location. Results demonstrate that such genetic data on parentage can be helpful not only in estimating contemporary population sizes, but also in providing additional information, not present in customary mark-recapture data, about possible extended movements of breeding individuals and the size of the pool of mates which they encounter. PMID- 11883885 TI - Mating frequency and mating system of the polygynous ant, Leptothorax acervorum. AB - Multiple mating by queens (polyandry) and the occurrence of multiple queens in the same colony (polygyny) alter patterns of relatedness within societies of eusocial insects. This is predicted to influence kin-selected conflicts over reproduction. We investigated the mating system of a facultatively polygynous UK population of the ant Leptothorax acervorum using up to six microsatellite loci. We estimated mating frequency by genotyping 79 dealate (colony) queens and the contents of their sperm receptacles and by detailed genetic analysis of 11 monogynous (single-queen) and nine polygynous colonies. Results indicated that 95% of queens were singly mated and 5% of queens were doubly mated. The corrected population mean mating frequency was 1.06. Parentage analysis of adults and brood in 17 colonies (10 monogynous, 7 polygynous) showed that female offspring attributable to each of 31 queens were full sisters, confirming that queens typically mate once. Inbreeding coefficients, queen-mate relatedness of zero and the low incidence of diploid males provided evidence that L. acervorum sexuals mate entirely or almost entirely at random. Males mated to queens in the same polygynous colony were not related to one another. Our data also confirmed that polygynous colonies contain queens that are related on average and that their workers had a mixed maternity. We conclude that the mating system of L. acervorum involves queens that mate near nests with unrelated males and then seek readoption by those nests, and queens that mate in mating aggregations away from nests, also with unrelated males. PMID- 11883886 TI - Contrasting evolutionary forces driving population structure at expressed sequence tag polymorphisms, allozymes and quantitative traits in white spruce. AB - Patterns of variation in quantitative characters and genetic markers were compared among six regional populations of white spruce [Picea glauca (Moench) Voss]. Although some phenotypic characters were correlated with latitude (r = 0.791), longitude (r = -0.796) and precipitation during the growing season (r = 0.789), variability at genetic markers was not correlated with geographical or bioclimatic variables, and followed neutral expectations. Estimates of genetic diversity and population differentiation for 14 allozymes (translated regions of coding genes) were essentially indistinguishable from those observed for 11 expressed sequence tag polymorphisms (ESTPs) from untranslated regions of coding genes. Variation among populations for quantitative traits such as eighth year height (Q(ST) = 0.082), thirteenth year height (Q(ST) = 0.069), total wood density (Q(ST) = 0.102) and date of budset (Q(ST) = 0.246), was greater than for allozymes (G(ST) = 0.014) and ESTPs (G(ST) = 0.019). These trends suggest a strong adaptive response in quantitative traits, contrasting to allozymes and ESTPs where no selective response could be detected and where populations appeared to be essentially in a migration-drift equilibrium. PMID- 11883887 TI - Pharmacokinetics and residues of ciprofloxacin and its metabolites in broiler chickens. AB - The pharmacokinetic properties of ciprofloxacin and its metabolites were determined in healthy chickens after single i.v. and oral dosage of 8 mg ciprofloxacin kg(-1) bodyweight. After i.v. and oral administration, the plasma concentration-time graph was characteristic of a two-compartment open model. Mean (SD) elimination half-life and mean residence time of ciprofloxacin in plasma were 8.84 (2.13) and 8.54 (1.64) hours, respectively, after i.v. administration and 11.89 (1.95) and 13.32 (2.65) hours, respectively, after oral administration. Mean maximal plasma concentration of ciprofloxacin was 2.63 (0.20) microg ml(-1), and the interval from oral administration until maximum concentration was 0.36 (0.07) hours. The mean oral bioavailability of ciprofloxacin was found to be 69.12 (6.95) per cent. Ciprofloxacin was mainly converted to oxociprofloxacin and desethyleneciprofloxacin. Considerable kidney, liver, muscle and skin + fat tissue concentrations of ciprofloxacin and its metabolites oxociprofloxacin and desethyleneciprofloxacin were found when ciprofloxacin was administered orally (8 mg kg(-1) on 3 successive days). It was estimated that mean tissue concentrations of ciprofloxacin and its metabolites ranging between 0.011 to 0.75 microg g(-1) persisted for 5 days. PMID- 11883888 TI - Influence of equine herpesvirus type 2 infection on monocyte chemoattractant protein 1 gene transcription in equine blood mononuclear cells. AB - Representational difference analysis (RDA) was used to compare gene expression in equine mononuclear cells either infected with equine herpesvirus-2 (EHV-2) or adsorbed with inactivated EHV-2. Seven clones identified in non-infected cells after three rounds of selective subtraction and enrichment for differentially expressed genes contained sequences homologous to equine monocyte chemoattractant protein 1 (MCP-1). This suggested that EHV-2 may down-regulate MCP-1 transcription in infected cells. These findings correlate well with similar findings described for human cytomegalovirus and support the view that EHV-2 may have the ability to modify the chemokine environment of infected cells. This may constitute an important feature of EHV-2 biology, because such an ability has the potential to compromise host defence mechanisms and predispose to infection with other pathogens. PMID- 11883889 TI - Glycosylated haemoglobin in dogs: study of critical difference value. AB - Measurement of glycated proteins can be of use in diagnosis and monitoring of diabetic dogs. Its use in monitoring can be facilitated by comparison of results with a reference interval derived from levels in normal dogs. In this study, a commercial immunoturbidometric assay was used to measure glycosylated haemoglobin in 15 normal dogs over a 5-week period. Following statistical analysis of the results a critical difference value of 0.38 per cent was obtained. PMID- 11883890 TI - A DNA vaccine encoding MPB83 from Mycobacterium bovis reduces M. bovis dissemination to the kidneys of mice and is expressed in primary cell cultures of the European badger (Meles meles). AB - Nucleic acid (DNA) vaccination against tuberculosis in the European badger (Meles meles) is one approach to addressing the escalating problem of bovine tuberculosis in Great Britain. The aim of vaccination is to reduce the burden of tuberculosis within the badger population and the shedding of Mycobacterium bovis to levels that would break the transmission of infection to cattle. To this end, the vaccine would be required to limit the amount of disseminated tuberculosis in the badger, especially dissemination to the kidney from where M. bovis can be shed in the urine. A promising candidate DNA vaccine encoding a 26 kDa major antigen (MPB83) of M. bovis was evaluated in a mouse model of disseminated M. bovis infection. Using the DNA vaccine, protection against infection of the kidney was found to be greater than that achieved with the current live vaccine, Bacille Calmette-Guerin (BCG). Kidney tissue and skeletal muscle from the badger was used to derive primary cell cultures in which to examine the expression of MPB83 following transfection with the DNA vaccine. Kidney cortex gave rise to a monotypic culture of epithelial cells whilst the muscle gave rise to a mixed culture of fibroblasts and myoblasts. During culture the myoblasts differentiated into multinucleated myotubes, verified by immunofluorescent detection of mammalian desmin. Successful expression of MPB83 by transfected epithelial and myotube cells was confirmed by immunofluorescence using a monoclonal antibody specific to the protein. These observations fulfil the early requirements for the development of a DNA vaccine for badger tuberculosis. PMID- 11883891 TI - Genetic typing of ruminant pestivirus strains from Northern Ireland and the Republic of Ireland. AB - A study was performed to investigate the genotypes and sub-groups of pestiviruses present in ruminants in Ireland. These comprised one ovine and eighteen bovine pestiviruses from Northern Ireland and six bovine pestiviruses from the Republic of Ireland. A 288 base pair (bp) portion of the 5'-non coding region (5'-NCR) from each of 25 pestiviruses collected over a period of 31 years was amplified by reverse-transcription-polymerase chain reaction (RT-PCR) and the product directly sequenced. From each pestivirus, nucleotide sequences corresponding to bases 130 to 374 of the 5'-NCR of NADL were aligned and compared with each other and with the corresponding sequences of a number of reference, field or vaccinal strains of BVDV types I and II, border disease virus and classical swine fever virus. All of the 25 sequenced pestiviruses were found to be BVDV type Ia. These were closely related to the constituent viruses of the 2 inactivated vaccines currently licensed for use in Northern Ireland and to recent bovine isolates from England. PMID- 11883892 TI - Effects of zinc and vitamin A supplements on plasma levels of thyroid hormones, cholesterol, glucose and egg yolk cholesterol of laying hens. AB - The effects of zinc and vitamin A supplementation to the diet on some blood metabolites were evaluated in Hisex brown laying hens from 56 weeks to 68 weeks of age. A total of 130 birds were divided into two main groups according to vitamin A treatment (0 and 3.44 mg retinyl acetate kg(-1) feed, respectively), each consisting of 65 hens. Hens in both of the main groups were then divided into five zinc treatment groups (0, 25, 50, 100 and 200 mg zinc kg diet(-1) respectively) of 13 hens each. It was observed that plasma T4, T3 and total cholesterol levels were affected by only zinc supplementation. While 100 and 200 mg Zn kg(-1) decreased plasma T4 level compared to control value, plasma T3 level was reduced by 100 mg Zn kg(-1) compared to groups fed less Zn. Adding 50 and 200 mg Zn kg(-1) to the diet increased plasma total cholesterol level in the birds compared to other groups. Vitamin A, zinc, and their interaction did not influence the concentration of plasma high-density lipoprotein (HDL)-cholesterol, glucose and egg yolk cholesterol in laying hens. PMID- 11883893 TI - Specific enzyme activities in bronchoalveolar lavage fluid as an aid to diagnosis of tracheobronchitis and bronchopneumonia in dogs. AB - Lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase and gamma-glutamyl transferase enzyme activities, and total protein (TP), calcium, inorganic phosphate, urea nitrogen (UN) and creatinine concentrations in bronchoalveolar lavage fluid were investigated for their relative importance in the diagnosis of respiratory diseases in dogs. Bronchoalveolar lavage (BAL) fluid was obtained from 26 dogs (20 with respiratory diseases and six controls) following anaesthesia with sodium pentothal. Enzyme activities and biochemical parameters were measured in BAL fluid. LDH and ALP levels were significantly increased in 12 dogs with bronchopneumonia, but not in eight dogs with tracheobronchitis. Insignificant and variable levels of TP and UN concentrations were found in both groups. It was concluded that LDH and ALP enzyme activities could be considered as pointers to pulmonary inflammation and/or damage while TP and UN measurements in BAL fluid may have a place in the identification of changes in respiratory and vascular permeability. PMID- 11883894 TI - Kinematic characteristics of Andalusian, Arabian and Anglo-Arabian horses: a comparative study. AB - The aim of this study was to compare the kinematic trot characteristics of three different breeds of horse: Andalusian (AN, n = 15), Arabian (AR, n = 7) and Anglo Arabian (AA, n = 5) using standard computer-assisted videography (25 Hz). Linear, temporal and angular parameters in fore- and hind limbs were analysed in six randomly selected strides per horse. Normalised angle-time diagrams along the complete stride were obtained for all joints angles in each breed and specific kinematic characteristics were detected graphically. AA horses displayed longer swing durations in both limbs ans a shorter angular range of motion (ARM) in scapula and pelvis inclination and in shoulder, hip and forelimb retraction protraction angles. At lift off, stifle and tarsal joint angles were more flexed. In general, only small differences were observed in AR horse kinematics when compared with the other 2 breeds. AN horses presented negative overtracking length, which was positive in AR and AA. In AN horses the elbow and carpal joints were more flexed at the moment of maximal elevation, elbow and fore-fetlock joints also exhibited a larger ARM due to a smaller angle at maximal flexion. In the hind limbs, tarsal, hind fetlock and retraction-protraction angles presented a larger ARM in AN horses due to greater maximal flexion in the tarsal and hind fetlock joints. Fore- and hind fetlocks were also more flexed in horses from this breed. In conclusion, differences between kinematic variables at the trot were observed in the three breeds studied here, mainly in forelimb joints. The most outstanding feature was the greater forelimb flexion recorded in AN horses than in the other breeds which is consistent with the elevated movements in this breed. In AA horses, the ARM of proximal joints involved in retraction protraction in both fore- and hind limbs was smaller. All the differences observed highlighted the idiosyncratic nature of the trot in each breed; this may influence the functional capacity of each breed. PMID- 11883895 TI - Morphological and quantitative study of the Leydig cells of pigs fed with anabolic doses of clenbuterol. AB - The effects of clenbuterol administered at anabolic doses on the testicular interstitium were studied in 30 pigs allocated to three experimental groups. The diet of two groups was supplemented with clenbuterol (Clb) (1 ppm), but whereas in the Clb+ group the treatment was given until slaughter (treatment period: 3 months), in the Clb- group the clenbuterol was withdrawn 2 weeks before slaughter (treatment period: 2-5 months); in the control group, the pigs were fed without clenbuterol. For histological procedures, a fractional sampling scheme was applied and routine techniques for light and transmission electron microscopy were used. The results of subjective morphology and morphometrics showed slight differences between the treated and the control groups. Conversely, the stereological results identified a prominent hyperplasia of the Leydig cells and ultrastructural analysis of these cells revealed a conspicuous increase in the organelles related to testosterone production, suggesting a functional activation of the interstitial cells in response to the clenbuterol treatment. PMID- 11883896 TI - Motility of the large intestine and flow of digesta in pigs. AB - The motor function of the large intestine of pigs is incompletely understood. Here the ileo-caecal-colonic motility is investigated by means of chronically implanted extraluminal strain gauge transducers and simultaneous videofluoroscopy in six pigs. Motility parameters were evaluated by computer and manually. The dominant feature of the ileal motility were aborally propagating giant contractions (velocity: 3.9 (0.7) cm sec(-1)) occurring at intervals of 7-12 minutes. They pushed the ileal digesta into the caecum. Despite a fed-state, migrating motor complexes occurred at intervals of 131.5 (8.1) minutes consisting of repetitive peristaltic waves. The motility of the caecum showed clustered contractions representing haustral movements. Transfer of caecal digesta and gas into the colon was caused by peristaltic contractions. The motility of the proximal colon was characterised by long peristaltic waves resulting in a rapid aboral transport of gas and a slow aboral flow of digesta. The propagation velocities along the centripetal and centrifugal loops of the colonic coil were 2.8 (0.6) and 5.7 (0.8) cm sec(-1), respectively. About half of the colonic waves were coordinated with the ileal giant contractions and the caecal peristaltic waves. The contraction parameters showed pronounced differences between the ileum and large intestine. The contraction rise time of the caecal and colonic contractions was about twice that of the ileal contractions (5.1 (0.2) and 4.4 (0.6) seconds versus 2.2 (0.1) seconds). Consequently, the maximal frequencies of the caecal and colonic contractions were about half compared with the ileal contractions (5.3 (0.4) and 6.1 (0.1) contractions min(-1) versus 11.8 (0.3) contractions min(-1)). Results show that the contractile patterns and motor functions of the individual intestinal segments differ markedly. PMID- 11883897 TI - Kinetic properties of p53 phosphorylation by the human vaccinia-related kinase 1. AB - The vaccinia-related kinase 1 (VRK1) protein is a nuclear Ser-Thr kinase that phosphorylates p53 in Thr18. We have determined the enzyme properties regarding its different substrates. VRK1 has a high affinity for ATP (K(m) 50 microM) and is thus saturated by the intracellular concentration of ATP in vivo. VRK1 uses preferentially magnesium, but is also functional with manganese and zinc. The VRK1 protein is autophosphorylated in multiple residues without effect on its activity. One autophosphorylated residue, T355, is within the VRK1 regulatory carboxy terminus. The kinase phosphorylates p53 with a K(m) of 1 microM and is well suited to respond to the variations of intracellular p53 concentration, which fluctuates as a response to different types of cellular stress. PMID- 11883898 TI - Partial characterization of a cerebral thyroid hormone-responsive protein. AB - The thyroid hormone-responsive protein (THRP) is expressed in rat cerebral tissue and has 83% overall sequence homology with c-Abl interactor protein, Abi-2, which is a substrate for the tyrosine kinase activity of c-Abl. Within the core region of the two proteins, the sequence similarity approaches 99%. To determine whether THRP is a rat homologue of Abi-2 or is a distinct protein with unique properties, the tissue distribution of THRP and Abi-2 mRNA's was examined using a sensitive ribonuclease protection assay and probes specific for THRP and Abi-2, respectively. The THRP mRNA content of cerebral tissue (1340.0 +/- 126.5 arbitrary units) was 2.3-fold higher than Abi-2 mRNA (581.3 +/- 73.7), while the ratio of hepatic content of THRP mRNA (209.0 +/- 49.1) to hepatic Abi-2 mRNA (2923.0 +/- 378.7) was only 0.07 (P < 0.004). Very low levels of Abi-2 mRNA, but not THRP mRNA, were also found in the heart and small intestine. Experiments with PC12 cells transfected with the full-length THRP cDNA and grown in the presence or absence of a tyrosine kinase inhibitor, along with experiments where PC12 cells were cotransfected with the THRP cDNA with or without the wild-type or mutant (tyrosine kinase deficient) c-Abl cDNA, showed that THRP is tyrosine phosphorylated; however, it is not a substrate for c-Abl. These studies demonstrate that THRP and Abi-2 have distinct tissue distribution and distinct biological properties. PMID- 11883899 TI - Both ALG-2 and peflin, penta-EF-hand (PEF) proteins, are stabilized by dimerization through their fifth EF-hand regions. AB - ALG-2 (apoptosis-linked gene-2 protein) and peflin are Ca(2+)-binding proteins and belong to the penta-EF-hand (PEF) protein family, which includes calpain, sorcin, and grancalcin. ALG-2 forms either a homodimer or a heterodimer with peflin like other PEF proteins. In this study, we found that the fifth-EF-hand (EF-5) regions of both ALG-2 and peflin are essential for dimerization and their stabilities. Exogenously expressed EF-5-deletion (DeltaEF-5) mutants of ALG-2 and peflin were unstable and were not detected in HEK293 cells by Western blotting. In a pulse--chase experiment, the DeltaEF-5 mutants were rapidly degraded, but they were stabilized by treatment with a proteasome inhibitor, MG132. In MG132 treated cells, DeltaEF-5 mutants were recovered in the insoluble fractions. Transient coexpression of ALG-2 increased the peflin level. These results indicate that the absence of a fifth EF-hand results in rapid degradation by the proteasome. On the other hand, stable expression of exogenous peflin decreased the amount of endogenous peflin. The amount of peflin that can dimerize with ALG 2 seems to be restricted in mammalian cells. PMID- 11883900 TI - Molecular evolution and structure--function relationships of the superoxide dismutase gene families in angiosperms and their relationship to other eukaryotic and prokaryotic superoxide dismutases. AB - This study assesses whether the phylogenetic relationships between SODs from different organisms could assist in elucidating the functional relationships among these enzymes from evolutionarily distinct species. Phylogenetic trees and intron positions were compared to determine the relationships among these enzymes. Alignment of Cu/ZnSOD amino acid sequences indicates high homology among plant sequences, with some features that distinguish chloroplastic from cytosolic Cu/ZnSODs. Among eukaryotes, the plant SODs group together. Alignment of the Mn and FeSOD amino acid sequences indicates a higher degree of homology within the group of MnSODs (>70%) than within FeSODs (approximately 60%). Tree topologies are similar and reflect the taxonomic classification of the corresponding species. Intron number and position in the Cu/Zn Sod genes are highly conserved in plants. Genes encoding cytosolic SODs have seven introns and genes encoding chloroplastic SODs have eight introns, except the chloroplastic maize Sod1, which has seven. In Mn Sod genes the number and position of introns are highly conserved among plant species, but not among nonplant species. The link between the phylogenetic relationships and SOD functions remains unclear. Our findings suggest that the 5' region of these genes played a pivotal role in the evolution of function of these enzymes. Nevertheless, the system of SODs is highly structured and it is critical to understand the physiological differences between the SODs in response to different stresses in order to compare their functions and evolutionary history. PMID- 11883901 TI - Nuclear UDP-glucuronosyltransferases: identification of UGT2B7 and UGT1A6 in human liver nuclear membranes. AB - We have demonstrated the subcellular localization of the human UDP glucuronosyltransferases (UGTs), UGT2B7 and UGT1A6, in endoplasmic reticulum (ER) and nuclear membrane from human hepatocytes and cell lines, by in situ immunostaining and Western blot. Double immunostaining for UGT2B7 and calnexin, an ER resident protein, showed that UGT2B7 was equally present in ER and nuclear membrane whereas calnexin was present almost exclusively in ER. Immunogold labeling of HK293 cells expressing UGT2B7 established the presence of UGT2B7 in both nuclear membranes. Enzymatic assays with UGT2B7 substrates confirmed the presence of functional UGT2B7 protein in ER, whole nuclei, and both outer and inner nuclear membranes. This study has identified, for the first time, the presence of UGT2B7 and UGT1A6 in the nucleus and of UGT2B7 in the inner and outer nuclear membranes. This localization may play an important functional role within nuclei: protection from toxic compounds and/or control of steady-state concentrations of nuclear receptor ligands. PMID- 11883902 TI - Functional consequences of the G235R mutation in liver arginase leading to hyperargininemia. AB - Hyperargininemia is a rare autosomal disorder that results from a deficiency in hepatic type I arginase. This deficiency is the consequence of random point mutations that occur throughout the gene. The G235R patient mutation has been proposed to affect the catalytic activity and structural integrity of the protein [D. E. Ash, L. R. Scolnick, Z. F. Kanyo, J. G. Vockley, S. D. Cederbaum, and D. W. Christianson (1998) Mol. Genet. Metab. 64, 243-249]. The G235R (patient) and G235A (control) arginase mutants of rat liver arginase have been generated to probe the effects of these point mutations on the structure and function of hepatic type I arginase. Both mutant arginases were trimeric by gel filtration, but the control G235A mutant had 56% of wild-type activity and the G235R mutant had less than 0.03% activity compared to the wild-type enzyme. The G235R mutant contained undetectable levels of tightly bound manganese as determined by electron paramagnetic resonance, while the G235A mutant had a Mn(II) stoichiometry of 2 Mn/subunit. Molecular modeling indicates that the introduction of an arginine residue at position 235 results in a major rearrangement of the metal ligands that compromise Mn(II) binding. PMID- 11883904 TI - The liver-specific human alpha(1)-microglobulin/bikunin precursor (AMBP) is capable of self-association. AB - alpha-1-Microglobulin (A1M) and bikunin are two plasma glycoproteins encoded by an alpha-1-microglobulin/bikunin precursor (AMBP) gene. Despite their lack of any structural or functional relationship, both A1M and bikunin originate from AMBP cleavage by a furin-like protease that releases the two mature molecules. The AMBP gene maintains a tight control over its expression by a unique enhancer, which is controlled by several hepatocyte-enriched nuclear factors; however, the mechanisms of regulation of the intracellular levels of the AMBP protein are currently unknown. We report the ability of the AMBP protein to self-associate and form a dimer in a yeast environment using the yeast two-hybrid system and an in vitro dimerization assay. We also show that the A1M protein binds to its precursor protein, AMBP, whereas bikunin does not. This observation warrants further investigations for a dimerization-dependent intracellular control that AMBP may be involved in. The relevance of AMBP dimerization and its possible biological significance are postulated. PMID- 11883905 TI - Random mutagenesis of the zinc-binding motif of betaine-homocysteine methyltransferase reveals that Gly 214 is essential. AB - Betaine-homocysteine S-methyltransferase (BHMT; EC2.1.1.5) is a zinc metalloenzyme that catalyzes the transfer of a methyl group from betaine to homocysteine to produce dimethylglycine and Met, respectively. This enzyme is a member of a family of zinc-dependent methyltransferases that use thiols or selenols as methyl acceptors and which contain the following motif: G[ILV]NCX(20, 100)[ALV]X(2)[ILV]GGCCX(3)PX(2)I. We recently reported that the three cysteine residues within this motif function as ligands to zinc in BHMT because changing any of them to alanine abolished zinc-binding and enzyme activity (A. P. Breksa, III, and T. A. Garrow, 1999, Biochemistry 38, 13991-13998). To determine if other amino acid residues in this motif were critical for enzyme function, the two regions defined by the motif in human BHMT, GVNCH(218) and VRYIGGCCGFEPYHI(307), were subjected to semirandom and random site-directed mutagenesis. Mutant enzymes were classified as either active or inactive based on their ability to complement the Met auxotrophy of Escherichia coli strain J5-3. The Gly residue at position 214 was found to be absolutely essential for complementation. The positions occupied by Gly297, Gly298, and Gly301 favored substitutions of small amino acids like Ala and Ser. We hypothesize that these Gly residues provide the necessary flexibility to the Zn-binding region to permit coordination of the metal. PMID- 11883903 TI - Toxicological consequences of differential regulation of cytochrome p450 isoforms in rat brain regions by phenobarbital. AB - Cytochrome P4502B is an isoform of cytochrome P450 (P450) that is induced by the anticonvulsant drug phenobarbital. Here, we demonstrate the constitutive expression and predominant localization of CYP2B in neurons of rat brain. Administration of phenobarbital to rats resulted in selective induction of P450 levels in cortex and midbrain, while other regions were unaffected. Immunohistochemical localization of P4502B in brains of phenobarbital treated rats revealed localization of P4502B in neuronal cells, most predominantly the reticular neurons in midbrain. The anticancer agent 9-methoxy-N(2) methylellipticinium acetate (MMEA) has been shown to exhibit preferential neuronal toxicity in vitro. Pretreatment of rats with phenobarbital potentiated the toxicity of intrathecally administered MMEA in vivo, as seen by the degeneration of reticular neurons. Thus, induction of P450 in selective regions of brain by phenobarbital would profoundly influence xenobiotic metabolism in these regions, especially in clinical situations where phenobarbital is coadministered with other psychoactive drugs/xenobiotics. PMID- 11883906 TI - A poplar plastocyanin mutant suitable for adsorption onto gold surface via disulfide bridge. AB - Aiming to achieve stable immobilization for a redox-active cupredoxin protein onto a gold substrate and its consequent molecular level monitoring by Scanning Tunnelling Microscopy (STM), we introduced a disulphide bridge within poplar plastocyanin, while avoiding the perturbation of its active site. We selected and modified residues Ile-21 to Cys and Glu-25 to Cys by structurally conservative mutagenesis. Optical absorption spectroscopy (UV-Vis), electron paramagnetic resonance (EPR), and resonance raman scattering (RRS) results indicate that the active site of the Ile21Cys, Glu25Cys plastocyanin (PCSS) to a large extent retains the spectroscopic properties of the wild-type protein. Furthermore, the redox midpoint potential of the couple CuII/CuI in PCSS, determined by cyclic voltammetry was found to be +348 mV close to the wild-type value. The STM images display self-assembled PCSS molecules immobilised onto gold substrate. Moreover, the full potentiostatic control of the electron transfer reaction during STM imaging, suggests that the adsorbed molecule maintains essentially its native redox properties. PMID- 11883907 TI - Dimethyl sulfoxide-induced conformational state of Na(+)/K(+)-ATPase studied by proteolytic cleavage. AB - Effects of dimethyl sulfoxide (Me(2)SO) on substrate affinity for phosphorylation by inorganic phosphate, on phosphorylation by ATP in the absence of Na(+), and on ouabain binding to the free form of the Na(+)/K(+)-ATPase have been attributed to changes in solvation of the active site or Me(2)SO-induced changes in the structure of the enzyme. Here we used selective trypsin cleavage as a procedure to determine the conformations that the Na(+)/K(+)-ATPase acquires in Me(2)SO medium. In water or in Me(2)SO medium, Na(+)/K(+)-ATPase exhibited after partial proteolysis two distinct groups of fragments: (1) in the presence of 0.1 M Na(+) or 0.1 M Na(+) + 3 mM ADP (enzyme in the E1 state) cleavage produced a main fragment of about 76 kDa; and (2) in the presence of 20 mM K(+) (E2 state) a 58 kDa fragment plus two or three fragments of 39-41 kDa were obtained. Cleavage in Me(2)SO medium in the absence of Na(+) and K(+) exhibited the same breakdown pattern as that obtained in the presence of K(+), but a 43-kDa fragment was also observed. An increase in the K(+) concentration to 0.5 mM eliminated the 43-kDa fragment, while a 39- to 41-kDa doublet was accumulated. Both in water and in Me(2)SO medium, a strong enhancement of the 43-kDa band was observed in the presence of either P(i) + ouabain or vanadate, suggesting that the 43-kDa fragment is closely related to the conformation of the phosphorylated enzyme. These results indicate that Me(2)SO acts not only by promoting the release of water from the ATP site, but also by inducing a conformation closely related to the phosphorylated state, even when the enzyme is not phosphorylated. PMID- 11883908 TI - Association of Cu,Zn-type superoxide dismutase with mitochondria and peroxisomes. AB - The subcellular localization of Cu,Zn-type superoxide dismutase (Cu,Zn-SOD) was investigated in rat tissues and cultured human fibroblasts. Subcellular fractionation, Nycodenz gradient centrifugation, and immunoblot analysis using specific antibodies showed that Cu,Zn-SOD was localized in cytosol, mitochondria, and peroxisomes of rat liver and brain. Treatment of highly purified mitochondria from rat liver with either Chaps or Triton X-100 released the bound Cu,Zn-SOD into supernatant fraction. Depolarization of mitochondria by inorganic phosphate and Ca(2+) released both Cu,Zn-SOD and cytochrome c from mitochondria. Digitonin also released Cu,Zn-SOD but not cytochrome c from mitochondria. Confocal immunofluorescence microscopy revealed that anti-Cu,Zn-SOD antibody in cultured human fibroblasts was found to colocalize with antibodies to Mn-SOD and PMP-70, markers of mitochondria and peroxisomes, respectively. Incubation of human Cu,Zn SOD with purified mitochondria resulted in their association. These results indicate that Cu,Zn-SOD associates with mitochondria and peroxisomes in various cell types such as those in brain, liver, and skin. PMID- 11883909 TI - Characterization of flavonoid--biomembrane interactions. AB - The flavonoids comprise a large group of polyphenolic compounds that are ubiquitous in vegetables, berries, and fruits, and they have been shown to possess antioxidative activity. The interactions between flavonoids and membranes composed of dipalmitoylphosphatidylcholine (DPPC) have been studied by means of noncovalent immobilized artificial membrane (IAM) chromatography. We have also investigated flavonoid-induced calcein release from fluid egg phosphatidylcholine (EPC) vesicles. Flavonoids with more hydroxyl groups showed longer retention delays in the IAM studies, suggesting stronger interactions between the flavonoids, which are rich in hydroxyl groups, and the DPPC membrane interface. We also observed an inverse correlation between the number of hydroxyl groups in the flavonoids and their capacity to induce calcein leakage through fluid EPC bilayer membranes (the more nonpolar flavonoids caused more calcein leakage). Rhamnetin and morin, however, both showed marked activity for the DPPC membrane interface and caused significant membrane leakage. Both polar and nonpolar forces were shown to have a significant impact on the flavonoid/biomembrane interactions. PMID- 11883912 TI - Genetic taste sensitivity to 6-n-propylthiouracil influences food preference and reported intake in preschool children. AB - Adult tasters of 6-n-propylthiouracil (PROP) are more sensitive to bitter taste and fattiness in foods, and often show lower acceptance of foods that are high in these taste qualities. This study hypothesized that PROP taster children would show lower acceptance of these same foods. Sixty-seven preschool children were classified as PROP tasters (N = 43) or nontasters (N = 24) using a suprathreshold screening solution. Children rated acceptance of 10 bitter and/or fat-containing foods using a 5-pt. facial scale. Parents completed a food frequency questionnaire to estimate their child's intake. Taster children showed lower acceptance of raw broccoli and American cheese (p < or = 0 x 05). Taster-girls showed lower acceptance of full-fat milk than nontaster-girls (p < or = 0 x 05). This effect was not seen in boys. Nontasters reported more daily intake of discretionary fats than tasters (p < or = 0 x 05), an effect largely due to nontaster-girls, in whom reported intake was 2--3 more servings per day than taster-girls, and boys of both groups. These data suggest that PROP taste sensitivity plays a role in acceptance of certain bitter cruciferous vegetables and cheese by young children. In addition, taster group differences in acceptance of full-fat milk and intake of discretionary fats seen in girls, suggest that gender-specific environmental factors might interact with genetics to influence fat preferences. PMID- 11883910 TI - Kinetic and regulatory properties of HK I(+), a modified form of the type I isozyme of mammalian hexokinase in which interactions between the N- and C terminal halves have been disrupted. AB - A modified form (HK I(+)) of rat Type I hexokinase (HK I) has been expressed. HK I(+) contains a centrally located polyalanine insert which, along with the known helical propensity of adjacent sequence, was expected to lead to alpha-helix formation, with resulting distension of the molecule and disruption of interactions between the N- and C-terminal halves. The properties of HK I(+) are consistent with this expectation and with previous proposals that (1) inhibition of HK I by Glc-6-P or its analogs and antagonism of this inhibition by P(i) result from competition of these ligands for a binding site in the N-terminal half of HK I, with resulting conformational changes propagated through interactions with the catalytic C-terminal half, and (2) binding of Glc-6-P to a site in the C-terminal half of HK I is obstructed by interactions between the halves, present in HK I but not HK I(+). PMID- 11883913 TI - Self-reported dietary restraint is associated with elevated levels of salivary cortisol. AB - Previous studies have found inconsistent relationships between restrained eating, dieting, and cortisol. The present study was designed to clarify the relationship between self-reported restrained eating and cortisol using multiple measures of dietary restraint. Eighty-five college-age women completed the Restraint Scale (RS) and the Cognitive Restraint Scale of the Three Factor Eating Questionnaire (TFEQ-R) and provided a saliva sample for analysis of cortisol. Both measures of restraint were positively associated with elevated levels of salivary cortisol, although the TFEQ-R was more strongly associated than the RS. Restrained eating, characterized by largely unsuccessful efforts to control eating, may lead to elevated cortisol levels. PMID- 11883914 TI - Stimulus satiation: effects of repeated exposure to foods on pleasantness and intake. AB - Frequent and repeated exposure to foods produces stimulus satiation or monotony. To explore further the nature of stimulus satiation, two experiments were conducted. Experiment 1 investigated the influence of initial pleasantness and frequency of intake on monotony. Tests showed that bread and butter was eaten more frequently but was liked less than chocolate. Therefore, normal-weight, healthy males were randomly assigned to either a chocolate condition (CC, N=13) or bread and butter condition (BC, N=16). All subjects received fixed amounts of the assigned food (67g/1473kJ of chocolate or 95g/1355kJ of bread and butter) every day for 22 days. On days 1, 8, 15 and 22 subjects consumed this food ad libitum. Pleasantness of taste and desire to eat chocolate declined significantly over time but no such changes were observed for bread and butter. Experiment 2 examined intake, pleasantness and desire to eat chocolate in 53 subjects over a 15 day period, with 3 conditions: control (CS: N=15), fixed (FS: N=20) and variable (VS: N=18). CS received no chocolate except on test days (days 1, 8 and 15), FS received 67g/1473kJ of chocolate daily and VS received increasing amounts of chocolate from 57g/1251kJ on day 1 to 86g/1888kJ by day 12. Pleasantness and desire to eat chocolate declined over time with this being more pronounced for F and V subjects. However, ad libitum intake increased over time. Both experiments demonstrated significant changes in pleasantness and desire to eat chocolate, but no commensurate decline in intake. Thus, although stimulus satiation occurred for subjective ratings of pleasantness and desire to eat chocolate, intake remained unaffected. This apparent dissociation between pleasantness and intake may reflect different processes underlying liking and wanting. PMID- 11883915 TI - Magnesium appetite in the rat. AB - Rats modify their ingestive behaviour to correct deficiencies of minerals such as sodium and calcium. Here, we examined the effect of magnesium deprivation on the ingestion of MgCl2 and other solutions. Male Sprague-Dawley rats were fed a nutritionally complete or magnesium-deficient diet and were then given 3.2, 10, 32, or 100mM MgCl2, 32mM CaCl2, 32mM NaCl, 10mM HCl, or 2.5mM saccharin, and their intake was measured for 24h in a two-bottle choice test with water. Within the first 5 min, magnesium-deprived subjects given 3.2, 32, or 100mM MgCl2 or 32mM CaCl2 drank significantly more of these solutions than did replete rats. In a separate study, rats fed replete, magnesium-deficient, or calcium-deficient diets were given a three-bottle choice between water, 32mM MgCl2, and 32mM CaCl2. The deprived rats preferred the solution that ameliorated their deficiency; for example, during the first 1h, the magnesium-deprived rats drank 3.1 +/- 0.5ml MgCl2 and 1.1 +/- 0.4ml CaCl2, whereas the calcium-deprived rats drank 1.8 +/- 0.5ml MgCl2 and 3.9 +/- 0.4ml CaCl2. Thus, magnesium deprivation leads to a compensatory appetite for magnesium, and the appetites for magnesium and calcium are distinct and specific. The rapid expression of magnesium appetite suggests that it depends in part on innate, gustatory factors. PMID- 11883916 TI - Rigid vs. flexible dieting: association with eating disorder symptoms in nonobese women. AB - The correlates of rigid and flexible dieting were examined in a sample of 188 nonobese women recruited from the community and from a university. The primary aim of the study was to test the hypothesis that women who utilize rigid versus flexible dieting strategies to prevent weight gain report more eating disorder symptoms and higher body mass index (BMI) in comparison to women who utilize flexible dieting strategies. The study sample included women who were underweight (29%), normal weight (52%), and overweight (19%). None of the women were obese, as defined by BMI>30. Participants were administered a questionnaire that measures Rigid Control and Flexible Control of eating. Body weight and height were measured and measures of eating disorder symptoms and mood disturbances were administered. Our results indicated that BMI was significantly correlated with rigid dieting and flexible dieting. BMI was controlled statistically in other analyses. The study found that individuals who engage in rigid dieting strategies reported symptoms of an eating disorder, mood disturbances, and excessive concern with body size/shape. In contrast, flexible dieting strategies were not highly associated with BMI, eating disorder symptoms, mood disturbances, or concerns with body size. Since this was a cross sectional study, causality of eating disorder symptoms could not be addressed. These findings replicate and extend the findings of earlier studies. These findings suggest that rigid dieting strategies, but not flexible dieting strategies, are associated with eating disorder symptoms and higher BMI in nonobese women. PMID- 11883917 TI - Food cravings and aversions during pregnancy: relationships with nausea and vomiting. AB - Food cravings and food aversions are common during pregnancy. A mechanism that may explain these changes in food preference is taste aversion learning. Accordingly, this study examined the temporal association between the first occurrences of nausea, vomiting, food cravings and food aversions during pregnancy. Ninety-nine women completed a questionnaire that asked about the occurrence, timing of first onset, duration, strength and targets of these symptoms. Nausea and vomiting were reported by 80% and 56% of the women, food cravings and aversions by 61% and 54% respectively. Although more women experienced both food cravings and aversions than either symptom alone, cravings and aversions were statistically unrelated. There was a significant positive correlation between week of onset of nausea and of aversions. In 60% of women reporting both nausea and food aversions, the first occurrence of each happened in the same week of pregnancy. No such association was found for cravings. These retrospective accounts provide good support for taste aversion learning as a mechanism for the development of some but not all food aversions during pregnancy. Prospective data are needed to confirm these temporal relationships and to assist understanding of the emergence of food cravings. PMID- 11883918 TI - Attitudes towards meat and meat-eating among adolescents in Norway: a qualitative study. AB - The aim of this study was to explore the concept of disgust in relation to meat and meat-eating. A sample of 30 high school students (aged 16 to 17 years), 10 urban females, 10 rural females, and 10 rural males, participated in the study. The participants' attitudes towards meat and meat-eating were investigated through interviews of a semi-structured nature and a short, confidential questionnaire. The study showed that disgust was solely related to red meat varieties and not to chicken. There were no vegetarians in our consumer sample, but red meat-eating was more common among males than females. Sensory attributes that were drivers of liking for meat were good taste, good smell and juiciness; these were described by both genders. All the females tended to characterise meat and meat-eating experiences negatively. Their associations were based on disgust, rather than distaste as found among males. Offensive attributes that the females attributed to meat were linked to the animals and their body parts, blood and raw meat, fibrous and chewy texture, fatty feeling in the mouth, and visible fat. Subjects with regular contact with farm animals displayed more relaxed attitudes towards animal production and showed no such disgust reactions. Females also tended to associate meat with "heavy" food that had negative impact on their bodies. They were also less content with their body appearance, dieted more than males, and tended to associate health (in the sense of fat consumption) and food intake to the wish for slim bodies. PMID- 11883919 TI - Reasons for eating: personal experiences in nutrition and anthropology. AB - Social, ecological, physiological and cognitive processes all influence choices among foods that cumulate in dietary intake. This broad research field is studied by nutritionists, agricultural economists and consumer researchers, specialists in ingestive behaviour, biosocial psychologists and cognitive anthropologists of food acceptance, sociologists and anthropologists of social roles of food and historians, folklorists, geographers and other cultural scholars of belief systems surrounding food research. Each discipline has its primary concerns, sometimes with other close fields. This workshop considered merits and mechanisms of inclusive research meetings, journals and books as physical units as well as separate workers and facilities for virtual conferences, documents and organizations. PMID- 11883920 TI - Nutrients epidemiology or healthy dietary practices? PMID- 11883921 TI - Appetite for integration: interdisciplinary careers for interdisciplinary areas. PMID- 11883922 TI - Interdisciplinarity, food, and power. PMID- 11883923 TI - Christine Wilson's presentation at the 1999 Food Choice Meeting in Toronto, Ontario, Canada. PMID- 11883924 TI - Personal perspectives and contemporary trends in nutritional anthropology. PMID- 11883925 TI - More personal experiences in nutrition and anthropology: a commentary on Christine Wilson. PMID- 11883926 TI - Food studies: interdisciplinary buffet and main course. PMID- 11883927 TI - Interdisciplinary work between nutrition and the social sciences: commentary on Christine Wilson's "reasons for eating: personal experiences in nutrition and anthropology". PMID- 11883928 TI - Are reasons required for eating? Comment on Wilson, Sobal, and Booth. PMID- 11883930 TI - Beta-1,4-galactosyltransferase and lactose synthase: molecular mechanical devices. AB - Recent structural investigations on the beta-1,4-galactosyltransferase-1 (Gal-T1) and lactose synthase (LS) have revealed that they are akin to an exquisite mechanical device with two well-coordinated flexible loops that are contained within the Gal-T1 catalytic domain. The smaller one has a Trp residue (Trp314) flanked by glycine residues. The larger one comprises amino acid residues 345 to 365. Upon substrate binding, the Trp314 side chain moves to lock the sugar nucleotide in the binding site, while the large loop undergoes a conformational change, masking the sugar nucleotide binding site, and creates (i) the oligosaccharide binding cavity; (ii) a protein-protein interacting site for the enzyme's partner, alpha-lactalbumin (LA); and (iii) a metal ion binding site. Only in conformation II do Gal-T1 and LA form the LS complex, enabling Gal-T1 to choose the new substrate glucose. LA holds and puts Glc right in the acceptor binding site of Gal-T1, which then maximizes the interactions with Glc, thereby making it a preferred acceptor for the LS reaction. The interaction of LA with Gal-T1 in conformation II also stabilizes the sugar-nucleotide-enzyme complex, kinetically enhancing the sugar transfer, even from the less preferred sugar nucleotides. The conformational change that masks the sugar nucleotide binding site can also be induced by the acceptor alone, thus making it possible for the protein to act as a specific lectin. PMID- 11883931 TI - Analysis of the role of RecQ helicases in RNAi in mammals. AB - The identity of mammalian genes involved in RNA interference (RNAi), the targeted sequence-specific mRNA degradation by double-stranded RNA (dsRNA), is poorly defined. Here we report the analysis of mice with null mutations of Wrn, Blm, and RecQ1 genes that are related to Mut-7 and Qde3, two genes essential for RNAi in Caenorhabditis elegans and quelling in Neurospora, respectively. Our results suggest that Wrn, Blm, and RecQ1 are not involved in sequence-specific mRNA degradation in mammals in response to dsRNA, suggesting potential differences in the mammalian RNAi pathway. PMID- 11883932 TI - Osmotic resistance of high-density erythrocytes in transglutaminase 2-deficient mice. AB - Transglutaminase 2 (TGase 2) is a Ca(2+)-dependent enzyme responsible for the posttransttranslational modification of proteins by transamidation of specific polypeptide-bound glutamine residues. Elevating the intracellular concentration of Ca(2+)-ions in human erythrocytes leads to the formation of cytoskeletal and cytoplasmic protein polymers. The Ca(2+)-dependent TGase 2-dependent cross linking activity has been proposed for its involvement in erythrocyte aging, by inducing irreversible modification of their cell shape and deformability. Accordingly, we found that high-density ("old") TGase 2(minus sign/minus sign) red blood cells (RBCs) were more resistant to osmotic stress-induced hemolysis than those from wild type mice. In addition, elevating the intracellular concentration of Ca(2+) by treatment of total RBCs with ionophore A23187 resulted in enhanced resistance of TGase 2-deficient erythrocytes compared to their normal counterpart. These findings indicate that TGase 2 may have a role in regulating structural flexibility of RBCs, possibly affecting their life span in physiopathological conditions, such as erythrocyte senescence, which are accompanied by increases in intracellular Ca(2+) concentration. PMID- 11883933 TI - Ubiquinone biosynthesis in rat liver peroxisomes. AB - The possibility that ubiquinone biosynthesis is present in rat liver peroxisomes was investigated. The specific activity of trans-prenyltransferase was 30% that of microsomes, with a pH optimum of around 8. trans-Geranyl pyrophosphate was required as a substrate and maximum activity was achieved with Mn(2+). Several detergents specifically inactivated the peroxisomal enzyme. The peroxisomal transferase is present in the luminal soluble contents, in contrast to the microsomal enzyme which is a membrane component. The treatment of rats with a number of drugs has demonstrated that the activities in the two organelles are subjected to separate regulation. Nonaprenyl-4-hydroxybenzoate transferase has about the same specific activity in peroxisomes as in microsomes and like the transferase activity, its regulation differs from the microsomal enzyme. The results demonstrate that peroxisomes are involved in ubiquinone biosynthesis, and at least two enzymes of the biosynthetic sequence are present in this organelle. PMID- 11883934 TI - Alternative splicing modulates subcellular localization of laforin. AB - Laforin is a dual-specificity phosphatase coded by the EPM2A gene defective in Lafora's progressive myoclonus epilepsy. We reported earlier that laforin is a cytoplasmic protein associated primarily with polyribosome. In the present study we characterized the expression of an EPM2A splice variant, named C-terISO, originating from the usage of a novel exon located in the 3'-untranslated region of exon 4 and encoding a laforin isoform containing unique sequences at its carboxyl terminus. Transfection studies demonstrate that, in addition to cytoplasm, the protein coded by C-terISO was targeted to the nucleus, a distinctive feature that was not observed for laforin coded by the major transcript of the EPM2A gene. The unique C-terminal sequence did not affect laforin's affinity for polysome, but sequestered nearly an equal amount of the protein into the nucleus. Our results are significant in light of the finding that laforin is an active phosphatase; therefore, isoforms targeted to different cellular compartments might dephosphorylate and regulate distinct cellular substrates. PMID- 11883935 TI - Transcriptional regulation of the human tumor suppressor p14(ARF) by E2F1, E2F2, E2F3, and Sp1-like factors. AB - The human ARF/INK4a locus encodes two cell cycle inhibitors, p16(INK4a) and p14(ARF), by using separate promoters. A variety of mitogenic stimuli upregulate ARF but a direct modulation at the transcriptional level has been reported only for E2F-1. We show here that the ARF promoter is strongly responsive also to E2F2 and E2F3, thus providing a strong support to their suggested role in the induction of apoptosis. Through the usage of both deletion mutants and/or site directed mutants, we surprisingly found that none of the four putative E2F consensus sites is strictly necessary for the upregulation of ARF expression, as a minimal deletion mutant, lacking all the putative E2F binding sites, is still transactivated by E2F. Moreover, our data suggest that the ARF promoter is regulated by E2F through both direct binding to the promoter sequences and indirectly, probably by being tethered to the ARF promoter by Sp1-like factors. PMID- 11883936 TI - Nuclear receptor and apoptosis initiator NGFI-B is a substrate for kinase ERK2. AB - NGFI-B is an inducible orphan nuclear receptor that initiates apoptosis. Growth factors such as EGF activate the MAP kinase ERK, whose activity may determine if a cell survives or undergoes apoptosis. EGF stimulation of cells leads to phosphorylation of threonine in NGFI-B. Thr-142 of NGFI-B is comprised in a consensus MAP kinase site and was identified as a preferred substrate for ERK2 (but not ERK1) in vitro. These results suggest that NGFI-B may be a molecular target for ERK2 signals and thereby a substrate for crosstalk between a growth factor survival pathway and an inducible regulator of apoptosis. PMID- 11883937 TI - A novel negative regulatory element in the human collagenase-3 proximal promoter region. AB - We have identified in the human collagenase-3 promoter a novel negative regulatory element, GAAAAGAAAAAG, designated AGRE (AG-Rich Element). The AGRE site functionality was characterized in human osteoarthritic (OA) chondrocytes as well as four cell lines. The cells were transfected with a plasmid consisting of the first 133 bp of the collagenase-3 promoter and its AGRE mutated or deleted derivatives. The absence of a functional AGRE site resulted in a statistically significant increase of the collagenase-3 basal transcription that was not affected by the collagenase-3 inducers IL-1beta and TGF-beta1. Two specific protein-AGRE binding complexes were detected by EMSA, and their presence depended on the physiological state of the cell. Indeed, normal chondrocytes and synovial fibroblasts and the four cell lines showed only a slower-migrating complex (complex 1). In OA chondrocytes, the type of complex discriminated two groups- the low-OA chondrocytes, showing low collagenase-3 basal levels and high inducibility of IL-1beta stimulation (complex 1), and the high-OA chondrocytes with high collagenase-3 basal levels and low IL-1beta inducibility (a faster migrating complex, designated complex 2). UV cross-linking revealed the presence of 48 and 97 kDa proteins in complex 1 and 27, 35, and 73 kDa proteins in complex 2. These findings suggest that the AGRE site plays a rate-limiting role in human collagenase-3 production. PMID- 11883938 TI - Genomic organization and regulation of a human 7-helix transmembrane receptor which is expressed in pulmonary epithelial cells and induced in hypoxia. AB - The genetic, regulatory, and tissue-specific characterization of G-protein coupled receptors is substantial since it contributes to the identification of the natural ligand which may influence basic physiological processes and cell function. Here we explored the genomic structure of a human orphan seven transmembrane receptor which presents the human homologue of a receptor which has been controversially identified as a rat adrenomedullin receptor subtype. Based on the cDNA sequence a 3.4 kb genomic DNA fragment was isolated. Sequencing of the fragment and comparison studies revealed an intron of 544 bp in the 5' untranslated region, followed by a second exon encoding the receptor protein of 404 amino acids. The gene is localized on chromosome 12q. The 5' regulatory region contains several SP1, AP2, and CAAT sites as well as hypoxia responsive elements (HRE) both in the 5' and 3' regulatory region. RT-PCR with intron spanning primers demonstrated mRNA signals in various tissues, especially in lung. Characterizing the histological expression pattern in lung sections by nonisotopic in situ hybridization, a strong signal of receptor mRNA was identified in pulmonary epithelial cells of bronchi and alveoli. Analysis of the two human pulmonary epithelial cell lines, H23 and A549, showed significant mRNA induction of this receptor subtype in hypoxia. PMID- 11883939 TI - ALG-2 interacts with the amino-terminal domain of annexin XI in a Ca(2+) dependent manner. AB - The apoptosis-linked protein ALG-2 is a Ca(2+)-binding protein that belongs to the penta-EF-hand protein family. ALG-2 forms a homodimer, a heterodimer with another penta-EF-hand protein, peflin, and a complex with its interacting protein, named AIP1 or Alix. By yeast two-hybrid screening using human ALG-2 as bait, we isolated a cDNA of a novel ALG-2-interacting protein, which turned out to be annexin XI. Deletion analysis revealed that ALG-2 interacted with the N terminal domain of annexin XI (AnxN), which has an amino acid sequence similar to that of the C-terminal region of AIP1/Alix. Using recombinant biotin-tagged ALG-2 and the glutathione S-transferase (GST) fusion protein of AnxN, the direct interaction was analyzed by an ALG-2 overlay assay and by real-time interaction analysis with a surface plasmon resonance (SPR) biosensor. The dissociation constant (K(d)) was estimated to be approximately 70 nM. The Ca(2+)-dependent fluorescence change of ALG-2 in the presence of the hydrophobicity fluorescent probe 2-p-toluidinylnaphthalene-6-sulfonate (TNS) was inhibited by mixing with GST-AnxN, suggesting that the Pro/Gly/Tyr/Ala-rich hydrophobic region in AnxN masked the Ca(2+)-dependently exposed hydrophobic surface of ALG-2. PMID- 11883940 TI - Expression of normal and mutant GFP-tagged y(+)L amino acid transporter-1 in mammalian cells. AB - Lysinuric protein intolerance (LPI; MIM 222700) is an autosomal recessive disorder characterized by defective transport of cationic amino acids lysine, arginine and ornithine. The defect is localized in the basolateral membrane of polar epithelial cells of the renal tubules and intestine. The SLC7A7 (solute carrier family 7, member 7) gene that encodes y(+)LAT-1 (y(+)L amino acid transporter-1) is mutated in LPI, and leads to the malfunction of the heterodimer composed of y(+)LAT-1 and 4F2hc (4F2 heavy chain) responsible for the system y(+)L amino acid transport activity at the membrane. In this study, the intracellular trafficking and membrane expression of wild type and four mutant y(+)LAT-1 proteins (LPI(Fin), G54V, 1548delC, W242X) was studied in two human cell lines by expressing green fluorescent protein (GFP) tagged proteins. Different SLC7A7 mutations influenced the trafficking of y(+)LAT-1 in the cells differently, as the wild type and missense mutant fusion proteins localized to the plasma membrane, while the frameshift and nonsense mutants sequestered to the cytoplasmic membranes, never reaching the target areas of the cell. PMID- 11883941 TI - Two splice variants of human PEX19 exhibit distinct functions in peroxisomal assembly. AB - PEX19 has been shown to play a central role in the early steps of peroxisomal membrane synthesis. Computational database analysis of the PEX19 sequence revealed three different conserved domains: D1 (aa 1--87), D2 (aa 88--272), and D3 (aa 273--299). However, these domains have not yet been linked to specific biological functions. We elected to functionally characterize the proteins derived from two naturally occurring PEX19 splice variants: PEX19DeltaE2 lacking the N-terminal domain D1 and PEX19DeltaE8 lacking the domain D3. Both interact with peroxisomal ABC transporters (ALDP, ALDRP, PMP70) and with full-length PEX3 as shown by in vitro protein interaction studies. PEX19DeltaE8 also interacts with a PEX3 protein lacking the peroxisomal targeting region located at the N terminus (Delta66aaPEX3), whereas PEX19DeltaE2 does not. Functional complementation studies in PEX19-deficient human fibroblasts revealed that transfection of PEX19DeltaE8-cDNA leads to restoration of both peroxisomal membranes and of functional peroxisomes, whereas transfection of PEX19DeltaE2 cDNA does not restore peroxisomal biogenesis. Human PEX19 is partly farnesylated in vitro and in vivo. The farnesylation consensus motif CLIM is located in the PEX19 domain D3. The finding that the protein derived from the splice variant lacking D3 is able to interact with several peroxisomal membrane proteins and to restore peroxisomal biogenesis challenges the previous assumption that farnesylation of PEX19 is essential for its biological functionality. The data presented demonstrate a considerable functional diversity of the proteins encoded by two PEX19 splice variants and thereby provide first experimental evidence for specific biological functions of the different predicted domains of the PEX19 protein. PMID- 11883942 TI - Isolation and characterization of a GnRH-like peptide from Octopus vulgaris. AB - Gonadotropin-releasing hormone (GnRH) is the key peptide in the hypothalamo hypophysial-gonadal axis, the core of regulation of reproduction in vertebrates. In this study, an octopus peptide with structural features similar to vertebrate GnRHs was isolated from brains of Octopus vulgaris. This peptide showed luteinizing hormone-releasing activity in quail anterior pituitary cells. A cDNA encoding the precursor protein was cloned. The RT-PCR transcripts were expressed in the supraesophageal and subesophageal brains, peduncle complex, and optic gland. The presence of the peptide in the different brain region was confirmed with enzyme-linked immunosorbent assay and time-of-flight mass spectrometric analysis. Immunoreactive neuronal cell bodies and fibers were observed in the subpedunculate lobe that controls the optic-gland activity. Optic gland nerves and glandular cells in the optic gland were immunostained. The isolated peptide may be octopus GnRH that contributes to octopus reproduction not only as a neurohormone but also as an endocrine hormone. PMID- 11883943 TI - Dioxin-induced adseverin expression in the mouse thymus is strictly regulated and dependent on the aryl hydrocarbon receptor. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a ligand for the ubiquitous, intracellular aryl hydrocarbon receptor (AhR), up-regulates the actin-modulating protein adseverin in mouse lymphoid tissues, a response that may be correlated to the immunotoxicity of TCDD. Here, by using chimeric mice with TCDD-responsive (AhR(+/+)) hematopoietic cells and TCDD-unresponsive (AhR(minus sign/minus sign)) thymic stroma, or the reverse, we show that TCDD-induced expression of adseverin in thymus is dependent on AhR expression in hematopoietic cells but not in stroma. The use of fetal thymic organ cultures also indicates that TCDD-induced expression of adseverin is confined to the thymocytes. The thymic stroma showed no induction of adseverin expression after TCDD exposure, although TCDD clearly activated the AhR in these cells, as indicated by the induction of CYP1A1. Adseverin was not induced in the thymus of normal adult C57BL/6 mice exposed to beta-estradiol or dexamethasone, two other agents, which also cause thymic atrophy. This further supports that adseverin induction is a specific gene regulatory effect by TCDD on thymocytes. PMID- 11883944 TI - Effects of a beta3-adrenergic agonist on glucose uptake and leptin expression and secretion in cultured adipocytes from lean and overweight (cafeteria) rats. AB - The increase in body and white adipose tissue weights induced by a high-fat diet were prevented by treatment with the beta3-adrenergic agonist Trecadrine. Plasma insulin levels were slightly elevated in overweight rats, while a decrease was observed in Trecadrine-treated groups. Insulin-dependent glucose uptake was impaired in adipocytes of the overweight rats in relation to lean animals. The beta3-adrenergic agonist induced an increase in insulin-stimulated glucose uptake by adipocytes as compared to the nontreated animals. In fact, Trecadrine treatment was able to restore to control values the impairment in insulin mediated glucose uptake induced by the cafeteria diet, suggesting that Trecadrine prevents the development of insulin resistance in overweight animals. Basal leptin secretion was increased in adipocytes of the overweight rats in relation to lean animals. Trecadrine treatment induced a decrease in basal leptin secretion compared to the untreated animals. Insulin-stimulated leptin secretion reached similar levels in adipocytes of the overweight rats as in lean animals. There was a trend for insulin-induced leptin secretion to be lower at 24 h in Trecadrine-treated rats, but it did not reach statistical significance. In conclusion, adipocytes of diet-induced overweight animals have a higher basal leptin secretion, which is reduced by treatment with Trecadrine. However, neither the cafeteria diet nor the Trecadrine treatment significantly alters the ability of adipocytes to increase leptin secretion in response to insulin. PMID- 11883945 TI - The effects of centrally administered apelin-13 on food intake, water intake and pituitary hormone release in rats. AB - Apelin is the recently identified endogenous ligand for the G-protein-coupled receptor, APJ. Preproapelin and APJ mRNA are found in hypothalamic regions known to be important in the regulation of food and water intake, and pituitary hormone release. The effects of intracerebroventricular (ICV) administration of pyroglutamylated apelin-13 on food and water intake and pituitary hormone release in rats were investigated. Apelin-13 had little effect on food intake, but dose dependently increased drinking behaviour and water intake at 1 h. Apelin-13 (10 nmol) increased water intake by up to sixfold compared to saline. Compared to saline control, apelin-13 (10 nmol) significantly increased plasma ACTH and corticosterone and decreased plasma prolactin, LH and FSH at 30 min. In vitro, apelin-13 stimulated the release of CRH and AVP from hypothalamic explants, but had no effect on NPY release. These results suggest that apelin may play an important role in the hypothalamic regulation of water intake and endocrine axes. PMID- 11883946 TI - A bisubstrate analog inhibitor of the carboxyltransferase component of acetyl-CoA carboxylase. AB - Acetyl-CoA carboxylase catalyzes the first committed step in the synthesis of long-chain fatty acids. The Escherichia coli form of the enzyme consists of a biotin carboxylase protein, a biotin carboxyl carrier protein, and a carboxyltransferase protein. In this report, the synthesis of a bisubstrate analog inhibitor of carboxyltransferase is described. The inhibitor was synthesized by covalently linking biotin to coenzyme A via an acyl bridge between the sulfur of coenzyme A and the 1'-N of biotin. The steady-state kinetics of carboxyltransferase are characterized in the reverse direction, in which malonyl CoA reacts with biocytin to form acetyl-CoA and carboxybiocytin. The inhibitor exhibited competitive inhibition versus malonyl-CoA and noncompetitive inhibition versus biocytin, with a slope inhibition constant (K(is)) of 23 +/- 2 microM. The bisubstrate analog has an affinity for carboxyltransferase 350 times higher than biotin. This suggests the inhibitor will be useful in structural studies, as well as aid in the search for chemotherapeutic agents that target acetyl-CoA carboxylase. PMID- 11883947 TI - Resveratrol activates membrane-bound guanylyl cyclase in coronary arterial smooth muscle: a novel signaling mechanism in support of coronary protection. AB - Resveratrol (RSVL), an edible polyphenolic stilbene, claims a myriad of cardiovascular benefits. However, the molecular underpinnings of such actions are poorly understood. Currently, in sheep coronary arteries (SCA), RSVL markedly (threefold) enhanced cGMP formation (t(1/2): 6.5 min; EC(50): 3 microM). This response was not abrogated by the phosphodiesterase inhibitor (IBMX, 0.5 mM), but was partly sensitive (20-30%) to either removal of the endothelium, treatment with the nitric oxide synthase-inhibitor (L-NMMA, 10 microM), or with the soluble GC (sGC)-inhibitor (ODQ, 10 microM). In membrane preparations from denuded SCA, either RSVL or the pGC agonist atrial natriuretic peptide (ANP, 0.1-1 microM) activated GC in the particulate, but not in the soluble, membrane fraction. By contrast, the nitric oxide donor, sodium nitroprusside (SNP, 1-10 microM), stimulated GC only in the soluble fraction. Further, pretreatment with RSVL partly desensitized the ANP response, but was additive to that of SNP. In arterial tension studies, RSVL relaxed PGF(2alpha)-precontracted denuded rings in a concentration-dependent manner, a response that was markedly enhanced (approximately 18 fold) in the presence of IBMX. Conversely, precontraction with phorbol ester, which also desensitizes pGC, blunted relaxations to RSVL but not to forskolin or SNP. These findings demonstrate that RSVL increases cGMP in coronary arteries, mostly by activation of pGC. This pathway triggers vasorelaxant responses that remain effective in endothelium-disrupted arteries. PMID- 11883948 TI - Relationship between the self-splicing activity and the solidity of the master domain of the Tetrahymena group I ribozyme. AB - The highly conserved P3-P7 domain of the Group I intron ribozymes is known to contain essential elements, such as the binding site for the cofactor guanosine, required for conducting the splicing reaction. We investigated the domain of the Tetrahymena intron ribozyme and its variants in order to clarify the relationship between its stability and function. We found that the destabilization of the P3 P7 domain facilitates the active structure formation at high magnesium ion concentrations where the formation is retarded for the wild type. The destabilized domain also increases K(GTP)(m) although this can be compensated by increasing the concentration of Mg(2+), indicating that the stable domain is required for establishing a tight guanosine binding site. The results suggest that the stability of the domain affects the rate-limiting step in the RNA folding pathway and also regulates the efficiency of the splicing reaction. PMID- 11883949 TI - ATP dependence of the SNARE/caveolin 1 interaction in the hippocampus. AB - The molecular mechanisms underlying the regulation of neurotransmission has been an open question for many years. Here, we have examined an interaction between caveolin1 and SNAREs (soluble N-ethylmalemide-sensitive factor attachment protein receptor) which may contribute to the cellular mechanisms underlying changes in synaptic strength. Previously, we reported that application of 4-aminopyridine to hippocampal slices resulted in a persistent potentiation of synaptic transmission and the induction of a short-lasting and specific 40-kDa complex composed of synaptosomal associated protein of 25 kDa (SNAP25) and caveolin1. We have characterized the binding properties of these proteins and observed that in vitro caveolin1 directly associates with both SNAP25 and syntaxin. Caveolin/SNARE interactions are enhanced in the presence of ATP by a mechanism that involves phosphorylation. While caveolin has been associated with cholesterol transport, signal transduction, and transcytosis, this study provides evidence that caveolin is also a SNARE accessory protein. PMID- 11883950 TI - Leucine zipper domain of HIV-1 gp41 interacted specifically with alpha-catenin. AB - Interactions between viral and cellular proteins could explain the molecular mechanisms behind the viral life cycle of HIV-1. The envelope protein gp41 of HIV 1 specifically interacted with alpha-catenin, not with beta-catenin. This interaction was shown by in vitro protein assay and in vivo transfected cell systems. Microinjection of the DNA expressing HIV-1 gp160 and alpha-catenin, into the HeLa cell, resulted in the colocalization of gp41 and alpha-catenin. Interestingly the noncleavable mutant of gp160 and alpha-catenin were found to be colocalized in the cell membrane. Mapping of the interaction sites between these two proteins revealed that the leucine zipper-like structure, located between the first and second alpha-helix domains from the carboxy terminus of HIV-1 gp41, interacted strongly with the carboxy terminus of alpha-catenin. PMID- 11883951 TI - Nucleotide-induced conformational changes of PMP70, an ATP binding cassette transporter on rat liver peroxisomal membranes. AB - Nucleotide-induced conformational changes of the 70-kDa peroxisomal membrane protein (PMP70) were investigated by means of limited-trypsin digestion. Rat liver peroxisomes preincubated with various nucleotides were subsequently digested by trypsin. The digestion products were subjected to immunoblot analysis with an anti-PMP70 antibody that recognizes the carboxyl-terminal 15 amino acids of the protein. PMP70 was initially cleaved in the boundary region between the transmembrane and nucleotide-binding domains and a carboxyl-terminal 30-kDa fragment resulted. The fragment in turn was progressively digested at the helical domain between the Walker A and B motifs. The fragment, however, could be stabilized with MgATP or MgADP. In contrast to MgATP, MgATP-gammaS protected whole PMP70 as well as the fragment. The 30-kDa fragment processed by trypsin was recovered in the post-peroxisomal fraction as a complex with a molecular mass of about 60 kDa irrespective of the presence of MgATP. These results suggest that PMP70 exists as a dimer on the peroxisomal membranes and the binding and hydrolysis of ATP induce conformational changes in PMP70 close to the boundary between the transmembrane and nucleotide binding domains and the helical domain between the Walker A and B motifs. PMID- 11883952 TI - Major adenine products from 2-methyl-1,4-naphthoquinone-sensitized photoirradiation at 365 nm. AB - In this article we report the isolation and characterization of major products of adenine in dinucleoside monophosphates upon 2-methyl-1,4-naphthoquinone (menadione)-sensitized UVA irradiation. Our results show that the major products form via the coupling between the menadione moiety and the exocyclic amino group of adenine. Similar reactions were not observed for cytosine. To our knowledge, this is the first report about the direct reaction between a DNA base and a photosensitizer under 365-nm ultraviolet light irradiation. Our results are consistent with previous observation showing that N(6) radical formed on adenine upon UVA irradiation. PMID- 11883954 TI - Potassium channel mRNAs with AU-rich elements and brain-specific expression. AB - GIRK2 (G protein-gated inwardly rectifying K(+) channel 2) located on the Down syndrome region 21q22.2 in humans has been reported to have several alternative transcripts and transcripts longer than 4 kb that do not have the poly-A tail. We sequenced GIRK2 transcripts with a long 3'-untranslated region (3'-UTR) containing multiple adenylate uridylate-rich elements (AREs) with the poly-A tail. In a 16-kb transcript, 28 AUUUA pentanucleotides, 9 AUUUUA hexanucleotides, 5 AUUUUUA heptanucleotides, and 3 UUAUUUA[U/A][U/A] nonanucleotides were found. Northern blot and in situ hybridization revealed abundant expression of the 16-kb transcripts in the rat brain despite no detectable signals in other tissues examined. The AREs have been reported to mediate the turnover of mRNAs encoding proteins regulating cellular proliferation/differentiation and body response to inflammatory and environmental stimuli. This is the first study indicating that ion channel transcripts have multiple AREs. PMID- 11883953 TI - Bcl-X(L) and calyculin A prevent translocation of Bax to mitochondria during apoptosis. AB - During many forms of apoptosis, Bax, a pro-apoptotic protein of the Bcl-2 family, translocates from the cytosol to the mitochondria and induces cytochrome c release, followed by caspase activation and DNA degradation. Both Bcl-X(L) and the protein phosphatase inhibitor calyculin A have been shown to prevent apoptosis, and here we investigated their impact on Bax translocation. ML-1 cells incubated with either anisomycin or staurosporine exhibited Bax translocation, cytochrome c release, caspase 8 activation, and Bid cleavage; only the latter two events were caspase-dependent, confirming that they are consequences in this apoptotic pathway. Both Bcl-X(L) and calyculin A prevented Bax translocation and cytochrome c release. Bcl-X(L) is generally thought to heterodimerize with Bax to prevent cytochrome c release and yet they remain in different cellular compartments, suggesting that their heterodimerization at the mitochondria is not the primary mechanism of Bcl-X(L)-mediated protection. Using chemical cross linking agents, Bax appeared to exist as a monomer in undamaged cells. Upon induction of apoptosis, Bax formed homo-oligomers in the mitochondrial fraction with no evidence for cross-linking to Bcl-2 or Bcl-X(L). Considering that both Bcl-X(L) and calyculin A inhibit Bax translocation, we propose that Bcl-X(L) may regulate Bax translocation through modulation of protein phosphatase or kinase signaling. PMID- 11883955 TI - High-order photobleaching of green fluorescent protein inside live cells in two photon excitation microscopy. AB - Combination of green fluorescent protein (GFP) and two-photon excitation fluorescence microscopy (TPE) has been used increasingly to study dynamic biochemical events within living cells, sometimes even in vivo. However, the high photon flux required in TPE may lead to higher-order photobleaching within the focal volume, which would introduce misinterpretation about the fine biochemical events. Here we first studied the high-order photobleaching rate of GFP inside live cells by measuring the dependence of the photobleaching rate on the excitation power. The photobleaching rate under one- and two-photon excitation increased with 1-power and 4-power of the incident intensity, respectively, implying the excitation photons might interact with excited fluorophore molecules and increase the probability of photobleaching. These results suggest that in applications where two-photon imaging of GFP is used to study dynamic molecular process, photobleaching may ruin the imaging results and attention should be paid in interpreting the imaging results. PMID- 11883956 TI - Probing the binding states of GDP to Cdc42 using urea interaction. AB - The inactive state of the small G protein Cdc42, the Cdc42.GDP.Mg(2+) ternary complex, was investigated using fluorescence, Mn(2+) substituted electron paramagnetic resonance, and (31)P nuclear magnetic resonance spectroscopy at various urea concentrations. The urea interaction with the protein was used to probe the binding state of GDP.Mg(2+) to Cdc42. Two binding states of the Cdc42.GDP.Mg(2+) ternary complex with different binding stability were observed. The two binding states were characterized by two sets of (31)P resonance of GDP phosphate groups, namely P(alpha) and P(beta), P('alpha), and P('beta). The high populated binding state I (P(alpha) and P(beta)) was more stable and less sensitive to the urea interaction. Yet the population of binding state II (P('alpha) and P('beta)) was lower, and the binding of GDP.Mg(2+) to Cdc42 in this state was more sensitive to the urea interaction. The release of GDP.Mg(2+) from the ternary complex in binding state II was faster than in state I. PMID- 11883957 TI - Deficiency of alpha-dystroglycan in muscle-eye-brain disease. AB - Alpha-dystroglycan is a component of the dystrophin-glycoprotein-complex, which is the major mechanism of attachment between the cytoskeleton and the extracellular matrix. Muscle-eye-brain disease (MEB) is an autosomal recessive disorder characterized by congenital muscular dystrophy, ocular abnormalities and lissencephaly. We recently found that MEB is caused by mutations in the protein O linked mannose beta1,2-N-acetylglucosaminyltransferase (POMGnT1) gene. POMGnT1 is a glycosylation enzyme that participates in the synthesis of O-mannosyl glycan, a modification that is rare in mammals but is known to be a laminin-binding ligand of alpha-dystroglycan. Here we report a selective deficiency of alpha dystroglycan in MEB patients. This finding suggests that alpha-dystroglycan is a potential target of POMGnT1 and that altered glycosylation of alpha-dystroglycan may play a critical role in the pathomechanism of MEB and some forms of muscular dystrophy. PMID- 11883958 TI - Inactivation of I(A) channels of frog skeletal muscle is modulated by ATP. AB - Whole cell, voltage clamp experiments were performed in vesicles derived from frog skeletal muscle plasma membranes to characterize the influence of ATP on the kinetic properties of fast inactivating K(+) currents (I(A)). I(A) was recorded in ATP-free solutions. Peak I(A) decayed with a time constant of 27 ms at large depolarizations. Steady state inactivation reached half maximal values at -66 mV. In the presence of ATP, these values were 196 ms and -41 mV, respectively, indicating a major effect of ATP on inactivation. In contrast, activation of I(A) was unaffected by ATP. The protein kinase C (PKC) inhibitors, H7 and staurosporine, greatly prevented the effects of ATP on inactivation. Inactivation remained unchanged by the protein kinase A inhibitor HA1004 or by the catalytic subunit of cAMP protein kinase. We conclude that ATP decreases inactivation of skeletal muscle I(A) and that this effect may be mediated by protein kinase C. PMID- 11883959 TI - Identification of the alternative splice products encoded by the human protein phosphatase inhibitor-1 gene. AB - We have isolated three major cDNA fragments of protein phosphatase inhibitor-1 from human brain and liver by RT-PCR. The 536 bp fragment encoded the wild-type of inhibitor-1 while two other fragments were alternative splice products of the inhibitor-1 gene, which was confirmed by partial genomic DNA sequencing. The 380 bp fragment encoded an in-frame 51-residue-deleted inhibitor-1, named inhibitor 1alpha, and the deletion occurred from residue 84 to 134 of inhibitor-1. The 316 bp fragment termed inhibitor-1beta was derived from an internal deletion of 536 bp fragment. This deletion resulted in an out of frame shift, allowing the 316 bp fragment that encoded the partial sequence of inhibitor-1. Based on the reported mRNA sequence of inhibitor-1 and evidence from our RT-PCR, we suggested that inhibitor-1beta consisted of 132 amino acids of which the N-terminal 61 amino acid sequences were identical to inhibitor-1 while the sequence after residue-61 was markedly different. PMID- 11883961 TI - Metformin effects on dipeptidylpeptidase IV degradation of glucagon-like peptide 1. AB - There is current interest in the use of inhibitors of dipeptidyl peptidase IV (DP IV) as therapeutic agents to normalize glycemic excursions in type 2 diabetic patients. Data indicating that metformin increases the circulating amount of active glucagon-like peptide-1 (GLP-1) in obese nondiabetic subjects have recently been presented, and it was proposed that metformin might act as a DP IV inhibitor. This possibility has been investigated directly using a number of in vitro methods. Studies were performed on DP IV enzyme from three sources: 20% human serum, purified porcine kidney DP IV, and recombinant human DP IV. Inhibition of DP IV hydrolysis of the substrate Gly-Pro-pNA by metformin was examined spectrophotometrically. Effects of metformin on GLP-1([7-36NH2]) degradation were assessed by mass spectrometry. In addition, surface plasmon resonance was used to establish whether or not metformin had any effect on GLP 1([7-36NH2]) or GLP-1([9-36NH2]) interaction with immobilized porcine or human DP IV. Metformin failed to alter the kinetics of Gly-Pro-pNA hydrolysis or GLP-1 degradation tested according to established methods. Surface plasmon resonance recordings indicated that both GLP-1([7-36NH2]) and GLP-1([9-36NH2]) show micromolar affinity (K(D)) for DP IV, but neither interaction was influenced by metformin. The results conclusively indicate that metformin does not act directly on DP IV, therefore alternative explanations for the purported effect of metformin on circulating active GLP-1 concentrations must be considered. PMID- 11883960 TI - Solution structure of intracellular signal-transducing peptide derived from human beta(2)-adrenergic receptor. AB - The structure of intracellular third loop peptide (betaIII-2: RRSSKFCLKEKKALK) was studied by CD and NMR spectroscopy. According to the CD study, this peptide forms a helix in the TFE solution. The three-dimensional molecular structure in TFE was determined by the 2D (1)H NMR spectroscopy. The structure consists of a positive charge cluster on the C-terminal side of the peptide chain. This part will be an active site of the peptide interacting with the G-protein. PMID- 11883962 TI - Clinical function after primary surgery for oral and oropharyngeal cancer: an 11 item examination. AB - The aim of this study was to record clinical function using an 11-item clinical examination and identify the main postoperative functional deficits. Of 132 consecutive patients undergoing surgery for previously untreated disease between January 1995 and June 1997, 130 were recruited in the study. An 11-domain clinical examination was made on the day before operation, and at 6 and 12 months afterwards. This examination assessed lip competence, tongue movement, oral mucosa, dental state, mouth opening, speech, drooling, diet, appearance, oral sensation and shoulder movement. Preoperatively there were deficits in natural dentition, consistency of diet and tongue protrusion. Postoperatively functional scores fell particularly for tongue movements, mouth opening, mucosa, dentition, speech, diet, appearance, lip sensation and tongue sensation. At 1 year, dental status, sensation and oral mucosa were particularly defective. Patients with large tumours, free tissue transfer, or adjuvant radiotherapy had the worst levels of function.A simple clinical examination provides a rapid assessment of function that can be used in conjunction with validated questionnaires to provide a more comprehensive evaluation of outcome. PMID- 11883963 TI - Health-related quality of life and clinical function after primary surgery for oral cancer. AB - Clinical function and health-related quality of life (HRQoL) are both important outcome parameters following surgery for oral and oropharyngeal cancer. The aim of this project was to explore the relationship between an 11-point clinical examination and HRQoL. Of 132 consecutive patients undergoing surgery for previously untreated disease between January 1995 and June 1997, 130 were recruited into the study. The University of Washington Quality of Life Questionnaire (UW-QoL) was completed by each patient on the day before operation and 6 and 12 months later. On each occasion the first author made an 11-point clinical examination. The main predictors of cumulative UW-QoL scores were tumour size, clinical functional score and type of operation. The trend was for a fall from preoperative levels at 6 months and then for a slight improvement at 1 year. The differential in respect of baseline function was present at all three time points in each patient group. This suggests that functional deficits at presentation persist following treatment. PMID- 11883964 TI - The role of open reduction and internal fixation in unilateral fractures of the mandibular condyle: a prospective study. AB - The purpose of this report is to present our experience in the management of unilateral condylar fractures between 1995 and 1998. This prospective study was carried out at a Regional Maxillofacial Unit and Teaching Hospital in the UK, and included 54 patients, of whom 32 attended for review. In all, 42 men and 12 women (age range 17-40 years) entered the study: 28 patients had unilateral fractures of the mandibular condyle alone; the remainder also had another mandibular fracture, invariably at the parasymphysis. Thirty-five of the patients (65%) had allegedly been assaulted. This is at variance with a recent study published by the British Association of Oral and Maxillofacial Surgeons, which showed that only 24% of facial injuries were the result of inter-personal violence. Open reduction and internal fixation of the mandibular condyle seems to carry low morbidity and may be of considerable functional benefit to the patient. PMID- 11883965 TI - Open reduction and internal fixation of fractured mandibular condyles by a retromandibular approach: surgical morbidity and informed consent. AB - Treatment of fractured mandibular condyles remains controversial. In this paper we present data collected prospectively on 42 fractured condyles treated by open reduction and internal fixation. The central issue, which is sometimes forgotten, is the informed consent of our patients. To this end we have looked at the surgical morbidity related to the retromandibular approach. PMID- 11883966 TI - Classification of surgical difficulty in extracting impacted third molars. AB - Few studies have attempted to analyse preoperative factors that complicate the surgical removal of impacted mandibular third molars. We studied this problem in two steps. We found that difficulty in extraction is associated with depth (depth is deep occlusal level: level C), ramus relationship/space available (ramus relationship/space available is no space: class 3), width of root (the width of the middle root is thicker than that of the neck and the roots do not separate, incomplete roots excluded: bulbous), or a combination of these factors. The index was tested in 20 patients whose extractions were difficult, and 24 in whom they were not. The new index has an odds ratio (relative risk) of 62.3 (95% confidence interval, 9.3-415.9), a sensitivity of 0.85 and a specificity of 0.92. We consider that the new index is superior to the conventional Pederson's index. PMID- 11883967 TI - Gastro-omental free flaps in oral and oropharyngeal reconstruction:surgical anatomy, complications, outcomes. AB - Free gastro-omental flaps can be used to reconstruct defects in the oral cavity after ablative cancer surgery. The omentum can provide as much bulk as required. The generous gastro-omental pedicle allows mobility. The gastric mucosal lining has the advantage that it produces mucus, does not carry hair follicles and is not prone to troublesome desquamation. This paper reviews the surgical anatomy of free gastro-omental flaps and presents a series of eight cases in which these flaps were used for oral and oropharyngeal reconstruction. PMID- 11883968 TI - Vascularization of the area between free grafts and irradiated graft beds in the neck in rats. AB - Inflammatory lesions of the vascular endothelium after preoperative radiotherapy often cause healing-delayed healing of free flaps in the irradiated graft bed. We investigated changes in neovascularization in the transition area between grafted tissues and irradiated tissues of the graft bed. We irradiated the neck(30 and 50 Gy total dose) in 102 Wistar rats and then grafted a free myocutaneous gracilis flap to the irradiated region of the neck 4 weeks later. We examined histologically the tissues of the graft, the transition area between the graft and the irradiated graft bed, and the graft bed. In contrast to control rats, the tissues in the irradiated animals showed a qualitatively reduced and a more irregular capillary distribution, with substantial fibrosis in the irradiated graft bed. We also found significant differences in vascularization and mean capillary lumen in the transitional zone between graft and graft bed in the irradiated rats compared with controls (P = 0.004 and P < 0.001, respectively). Both number and diameter of capillaries were reduced in the irradiated graft bed tissue. The graft failed to improve vascularization in the transitional zone between graft and irradiated tissue, so we conclude that it is the vascularization status of the bed tissue rather than that of the transplant tissue that is the limiting factor for graft healing. PMID- 11883969 TI - An extreme case of cherubism. AB - We describe an 8-year-old boy who presented with severe facial swelling. This progressed rapidly and 17 months later he died of gastrointestinal and pulmonary infections. The diagnosis was initially brown tumour associated with hyperparathyroidism, but this was revised in the light of laboratory investigations that were within the reference ranges, and normal appearance of the parathyroids on exploration to that of an extreme case of cherubism that behaved in a locally aggressive manner. PMID- 11883970 TI - The diagnosis and consequences of Stickler syndrome. AB - The objective was to study the expressivity of Stickler syndrome in affected children and adults in the UK and to highlight issues for improving early diagnosis, treatment and counselling. A postal questionnaire survey of the 216 members of the Stickler Syndrome Support Group was carried out. Of the 153 (71%) who responded to the questionnaire, 48 (61%) of adults and 15 (20%) of children had experienced retinal detachment; 36 (49%) of the children and 18 (23%) of the adults were born with a cleft palate. Only 5 (7%) of the children and none of the adults had been diagnosed by a cleft surgeon, although 23 (31%) of the children had been diagnosed originally as having Pierre-Robin sequence. Only a third of the adults had been given any genetic counselling. Stickler syndrome is an under diagnosed condition with profound consequences, particularly with respect to vision. Earlier diagnosis by the cleft team may help to reduce suffering and increase awareness of the condition. PMID- 11883971 TI - Routine follow-up after periradicular operations: current practice in UK hospitals. AB - Questionnaires were circulated to all Fellows of the British Society of Oral and Maxillofacial Surgeons in 1999, 75% of whom replied (n = 194). There was a wide range of responses for both the timing of review appointments and the taking of radiographs. Most patients were first followed up 1 week after operation (46%), but 38% were reviewed in the second week. Five surgeons did not review patients at all and two did not review until 6 or 9 months, respectively. Nearly two thirds routinely offered a second follow-up appointment but only 14% offered more than two, the maximum being seven. Most arranged postoperative radio-graphs but the timing ranged from immediately postoperatively to 1 year after the procedure. Less than one-third requested a second postoperative radiograph between 1 month and 1 year. The largest disparity was in the time of discharge to the general dental practitioner, which ranged from immediately to 5 years, the most popular time of discharge being at 3 months. The wide variations may reflect unnecessary recall of patients and misuse of valuable clinical time. PMID- 11883972 TI - Differential diagnosis and treatment of autosomal dominant osteosclerosis of the mandible. AB - A 20-year-old patient who presented with concerns about her large mandible was found to have a generalized mild cortical sclerosis. She was treated successfully with staged orthognathic surgery, despite the dense sclerosis of the jaws. PMID- 11883974 TI - Craniocervical necrotizing fasciitis in Ile-Ife, Nigeria. AB - Sixteen cases of necrotizing fasciitis were seen at the Obafemi Awolowo University Teaching Hospital, Ile-Ife, Nigeria from 1990 to 2000. Primary craniocervical involvement was recorded in seven patients (five men and two women). The clinical records of five patients were sufficiently detailed to allow us to report their age, aetiology, predisposing illness, clinical features, complications, management regimen and outcome. The patients were aged 30-75 years and in four of them odontogenic infections were the cause of the condition. Hypertension, diabetes mellitus and obesity were the underlying systemic diseases in three cases and the body/angle region of the mandible was the predominant site of the infection on the face. All five cases had involvement of the neck. Mediastinal extension was recorded in three cases. Two patients had complications: one had septicaemia and renal failure and the other developed bone necrosis. Pre-existing ill health, old age, late surgical intervention, and mediastinal and thoracic extension of infection were responsible for the only death. Treatment involved frequent and multiple surgical debridement, aggressive antimicrobial treatment and control of systemic disease. Early recognition, prompt surgical intervention, and aggressive antimicrobial treatment are essential to minimize morbidity and mortality. Rapid progression of infection, financial constraints, delayed referrals from rural clinics and distance to the tertiary hospital caused problems. PMID- 11883973 TI - Postoperative sore throat after routine oral surgery: influence of the presence of a pharyngeal pack. AB - A randomized clinical trial was conducted to investigate the effect of the presence of a pharyngeal pack during endotracheal anaesthesia on the incidence of postoperative sore throat. The patients were anaesthetized with fentanyl, thiopentone and atracurium. Thirty-six patients were anaesthetized without placement of pharyngeal packs while 26 patients had pharyngeal packs inserted. There were no significant differences in the incidence or severity of sore throat postoperatively in the two groups (P=0.23). These results contradict previous studies, which showed an increase in the incidence of postoperative sore throat after the use of pharyngeal packs. PMID- 11883975 TI - Proinflammatory cytokines and arthroscopic findings of patients with internal derangement and osteoarthritis of the temporomandibular joint. AB - This study investigated the correlations between the concentrations of proinflammatory cytokines in synovial fluid and the degree of synovitis on the one hand, and the degree of degeneration of articular cartilage on the other hand, in patients with internal derangement and osteoarthritis of the temporomandibular joint. We measured the concentrations of interleukin-1beta (IL 1beta), tumour necrosis factor-alpha (TNF-alpha), IL-6 and IL-8 in synovial fluid and the degree of arthroscopic synovitis and degeneration of articular cartilage in 37 joints with internal derangement and osteoarthritis. The correlations between the concentration of each cytokine and the score of each arthroscopic feature were analysed statistically. The detection rates of IL-1beta,TNF-alpha, IL-6 and IL-8 were 57%, 78%, 89% and 70%, respectively. There was a positive correlation between the IL-6 concentration and the synovitis score (P = 0.02). Measurement of IL-6 in synovial fluid might be useful as an indicator of the extent of synovitis. PMID- 11883976 TI - Importance of the changes in joint effusion shown by magnetic resonance imaging before and after arthroscopic lysis and lavage of the temporomandibular joint. AB - We investigated the changes in the amount of joint effusion estimated from T2 weighted magnetic resonance imaging (MRI) before and after arthroscopic lysis and lavage of the temporomandibular joint (TMJ). We studied 29 consecutive patients, each with internal derangement and osteoarthritis in one TMJ. Before operation, the MRI showed joint effusions in 22 of the patients (76%). After operation, the amount of the effusion decreased in 16 and increased in 2 patients. Effusions developed postoperatively in four of the seven patients who had no effusion before operation. In four of the six patients in whom the effusion increased, the symptoms had almost resolved by the time the MRI was taken. There was no significant correlation between changes in the amount of joint effusion and the clinical condition of the patients before and after the operation. In conclusion, changes in the amount of joint effusion in the TMJ are not related to the patient's clinical condition. PMID- 11883977 TI - Study by finite element method of the mechanical stress of selected biodegradable osteosynthesis screws in sagittal ramus osteotomy. AB - We tested the stability of the bilateral sagittal split osteotomy using four resorbable osteosynthesis screws (the PLLA screw introduced by Harada and Enomoto, the Isosorb screw, the BioSorbFX screw and the Lactosorb screw) which are all currently in clinical use. The distribution of stress in both the bicortically inserted screws and the adjacent bone of a computer-generated mandible was recorded by the three-dimensional finite element method. The stress of the materials under investigation was postulated to have reached threshold values for stability, and maximum chewing forces of 132 N (Harada and Enomoto), 117 N (Isosorb), 115 N (BioSorbFX) and 46.4 N (Lactosorb) were determined. As far as the postoperative chewing forces were concerned, all four screws were sufficiently stable at the osteotomy gap. Finite element modelling seems to be an appropriate method of investigating these clinical issues when the mechanical stress both in implants and in the adjacent bone is taken into account. PMID- 11883978 TI - Intermaxillary fixation screws and tooth damage. PMID- 11883979 TI - Iatrogenic injury caused by intermaxillary fixation screws. PMID- 11883980 TI - Re: Zakrzewska JM. Cluster headache: review of the literature. Br J Oral Maxillofac Surg 2001; 39: 103-113. PMID- 11883981 TI - Re: Holmes et al. Percutaneous osteosynthesis of the zygomatic buttress. Br J Oral Maxillofac Surg 2001; 39: 286-288. PMID- 11883982 TI - Conserving the inferior alveolar nerve during resection of the mandible. PMID- 11883983 TI - The incidence of maxillofacial injuries sustained during the May Day riots -- the experience of the Accident and Emergency Unit at the University College London Hospital. PMID- 11883984 TI - European Rolling Training Programme in Oral & Maxillofacial surgery 'Surgery for Facial Deformity -- an update' Manchester, 31 August - 2 September 2001. PMID- 11883985 TI - I, zombie. AB - Certain recent philosophical theories offer the prospect that zombies are possible. These theories argue that experiential contents, or qualia, are nonphysical properties. The arguments are based on the conceivability of alternate worlds in which physical laws and properties remain the same, but in which qualia either differ or are absent altogether. This article maintains that qualia are, on the contrary, physical properties in the world. It is shown how, under the burden of the a posteriori identification of qualia with physical properties, a reasoned choice can be made between the two types of theories which ultimately favors materialism and rejects zombies. PMID- 11883986 TI - Consciousness and control in task switching. AB - Participants were required to switch among randomly ordered tasks, and instructional cues were used to indicate which task to execute. In Experiments 1 and 2, the participants indicated their readiness for the task switch before they received the target stimulus; thus, each trial was associated with two primary dependent measures: (1) readiness time and (2) target reaction time. Slow readiness responses and instructions emphasizing high readiness were paradoxically accompanied by slow target reaction time. Moreover, the effect of task switching on readiness time was an order of magnitude smaller then the (objectively estimated) duration required for task preparation (Experiment 3). The results strongly suggest that participants have little conscious awareness of their preparedness and challenge commonly accepted assumptions concerning the role of consciousness in cognitive control. PMID- 11883987 TI - Emotion and cognition: feeling and character identification in dreaming. AB - This study investigated the relationship between dream emotion and dream character identification. Thirty-five subjects provided 320 dream reports and answers to questions on characters that appeared in their dreams. We found that emotions are almost always evoked by our dream characters and that they are often used as a basis for identifying them. We found that affection and joy were commonly associated with known characters and were used to identify them even when these emotional attributes were inconsistent with those of the waking state. These findings are consistent with the finding that the dorsolateral prefrontal cortex, associated with short-term memory, is less active in the dreaming compared to the wake brain, while the paleocortical and subcortical limbic areas are more active. The findings are also consistent with the suggestion that these limbic areas have minimal input from the dorsolateral prefrontal cortex in the dreaming brain. PMID- 11883988 TI - On the neural correlates of object recognition awareness: relationship to computational activities and activities mediating perceptual awareness. AB - Based on theoretical considerations of Aurell (1979) and Block (1995), we argue that object recognition awareness is distinct from purely sensory awareness and that the former is mediated by neuronal activities in areas that are separate and distinct from cortical sensory areas. We propose that two of the principal functions of neuronal activities in sensory cortex, which are to provide sensory awareness and to effect the computations that are necessary for object recognition, are dissociated. We provide examples of how this dissociation might be achieved and argue that the components of the neuronal activities which carry the computations do not directly enter the awareness of the subject. The results of these computations are sparse representations (i.e., vector or distributed codes) which are activated by the presentation of particular sensory objects and are essentially engrams for the recognition of objects. These final representations occur in the highest order areas of sensory cortex; in the visual analyzer, the areas include the anterior part of the inferior temporal cortex and the perirhinal cortex. We propose, based on lesion and connectional data, that the two areas in which activities provide recognition awareness are the temporopolar cortex and the medial orbitofrontal cortex. Activities in the temporopolar cortex provide the recognition awareness of objects learned in the remote past (consolidated object recognition), and those in the medial orbitofrontal cortex provide the recognition awareness of objects learned in the recent past. The activation of the sparse representation for a particular sensory object in turn activates neurons in one or both of these regions of cortex, and it is the activities of these neurons that provide the awareness of recognition of the object in question. The neural circuitry involved in the activation of these representations is discussed. PMID- 11883989 TI - Evidence for preserved representations in change blindness. AB - People often fail to detect large changes to scenes, provided that the changes occur during a visual disruption. This phenomenon, known as "change blindness," occurs both in the laboratory and in real-world situations in which changes occur unexpectedly. The pervasiveness of the inability to detect changes is consistent with the theoretical notion that we internally represent relatively little information from our visual world from one glance at a scene to the next. However, evidence for change blindness does not necessarily imply the absence of such a representation---people could also miss changes if they fail to compare an existing representation of the pre-change scene to the post-change scene. In three experiments, we show that people often do have a representation of some aspects of the pre-change scene even when they fail to report the change. And, in fact, they appear to "discover" this memory and can explicitly report details of a changed object in response to probing questions. The results of these real world change detection studies are discussed in the context of broader claims about change blindness. PMID- 11883990 TI - Can humans perceive their brain states? AB - Although the brain enables us to perceive the external world and our body, it remains unknown whether brain processes themselves can be perceived. Brain tissue does not have receptors for its own activity. However, the ability of humans to acquire self-control of brain processes indicates that the perception of these processes may also be achieved by learning. In this study patients learned to control low-frequency components of their EEG: the so-called slow cortical potentials (SCPs). In particular "probe" sessions, the patients estimated the quality of the SCP shift they had produced in the preceding trial. The correspondence between the recorded SCP amplitudes and the subjective estimates increased with training. The ability to perceive the SCPs was related to the ability to control them; this perception was not mediated by peripheral variables such as changes in muscle tonus and cannot be reduced to simple vigilance monitoring. These data provide evidence that humans can learn to perceive the neural activity of their brain. Alternative interpretations are discussed. PMID- 11883991 TI - Suppression of EEG gamma activity may cause the attentional blink. AB - The attentional blink (AB) is an impairment of attention, which occurs when subjects have to report a target stimulus (T2) following a previous target (T1) with a short delay (up to 600 ms). Theories explaining the AB assume that processing of T2 is more vulnerable to decay or substitution, as long as attention is allocated to T1. Existing models of the AB, however, do not account for the fact that T2 detection accuracy reaches the minimum when T2 is presented after about 300 ms and not immediately following T1. Therefore, a new model is suggested, which is based on chronometrical considerations together with recent neurophysiological findings concerning the relation between the P3 event-related potential and the AB, the interaction between P3 and gamma oscillations, and the significance of the early evoked gamma band response. We hypothesize that suppression of the early gamma response to T2, accompanying the P3 related to T1, causes the AB. PMID- 11883992 TI - Effects of the benzodiazepine lorazepam on monitoring and control processes in semantic memory. AB - Lorazepam has been repeatedly shown to induce memory impairments. The effects of this benzodiazepine on the processes involved in the strategic regulation of memory accuracy have not as yet been explored. An experimental procedure that delineates the role of monitoring and control processes was used. Fifteen lorazepam and 15 placebo subjects were examined using a semantic memory task that combined both a forced- and a free-report option and a no-incentive and an incentive condition. Memory accuracy was lower in the lorazepam than in the placebo group. Lorazepam impaired control sensitivity (the extent to which volunteering of answers is affected by the confidence judgments). While the absolute aspect of monitoring was impaired (calibration scores), both the discriminative aspect (the ability to distinguish between correct and incorrect answers) and the response criterion setting (the confidence threshold set for volunteering a report) were spared. The pharmacological dissociation between monitoring effectiveness and control sensitivity indicates that these two components involve distinct processes. PMID- 11883994 TI - Stress proteins in disease: metabolism on a knife edge. PMID- 11883995 TI - The adjunctive value of routine biochemistry in nutritional assessment of hospitalized patients. AB - Nutritional screening and assessment of patients are not part of the routine procedures at hospital admission, although a large percentage of patients is malnourished. Similarly, nutritional status of patients and dietary intake is usually not monitored during hospitalization. In contrast a great number of laboratory investigations for screening purposes accompanies most, if not all, hospital admissions. Several of those routine markers carry important nutritional information and could convey several aspects of patients' nutritional requirements. This review proposes that laboratories in particular could provide a service for the nutritional interpretation of available routine data which would help clinicians to focus on nutrition related problems. The nutritional meaning of basic physical characteristics (age, sex, height, and weight), urinary excretion of ketone bodies, urea and creatinine, and serum concentrations of urea, phosphate, iron and albumin are discussed in detail and the assessment of protein malnutrition, metabolism, and requirements is emphasized. Finally, a proposed sample layout for the nutritional interpretation of routinely available biochemical and basic physical data is presented. PMID- 11883996 TI - Malnutrition in hospitalized elderly patients: when does it matter? AB - BACKGROUND: Identifying the individual effects of acute illness and malnutrition on elderly patient outcome and the timing of nutritional support is still an important challenge for modern medicine. OBJECTIVE: The aims of this study were to assess the practical significance of serum albumin concentrations following acute illness as a measure of nutritional status in ageing patients and also to review recently published studies related to this field. DESIGN: Consecutive stroke patients had their nutritional status assessed from anthropometric, haematological and biochemical data during the hospital stay. Predicted energy needs and daily in-hospital energy intake were also studied in a subgroup of 24 acute stroke patients and 24 age and sex-matched hospitalized non-stroke patients. A multivariate analysis was used to measure the amount of variance in serum albumin concentrations explained by nutritional and non-nutritional clinical variables. RESULTS: Serum albumin concentrations deteriorated steadily during the study period and there was an increase in the amount of variance in the serum albumin explained by nutritional variables between admission and week 4 of the hospital stay. Almost all patients studied were in negative energy balance during hospitalization. Evidence is provided which links low serum albumin concentrations with clinical outcomes during the hospital stay and immediately following discharge. That nutritional supplementation started one week as opposed to immediately following acute illness, and continued during the convalescent period, can improve serum albumin concentrations during the hospital stay. CONCLUSION: Poor nutritional status following acute illness in ageing patients may be of more prognostic significance and amenable to therapy later on during the course of hospitalization. PMID- 11883997 TI - Urea synthesis in patients with chronic pancreatitis: relation to glucagon secretion and dietary protein intake. AB - BACKGROUND & AIMS: Up-regulation of urea synthesis by amino acids and dietary protein intake may be impaired in patients with chronic pancreatitis (CP) due to the reduced glucagon secretion. Conversely, urea synthesis may be increased as a result of the chronic inflammation. The aims of the study were to determine urea synthesis kinetics in CP patients in relation to glucagon secretion (study I) and during an increase in protein intake (study II). METHODS: In study I, urea synthesis rate, calculated as urinary excretion rate corrected for accumulation in total body water and intestinal loss, was measured during infusion of alanine in 7 CP patients and 5 control subjects on spontaneous protein intake. The functional hepatic nitrogen clearance (FHNC), i.e. urea synthesis expressed independent of changes in plasma amino acid concentration, was calculated as the slope of the linear relation between urea synthesis rate and plasma alpha -amino nitrogen concentration. In study II, 6 of the patients of study I had urea synthesis and FHNC determined before and after a period of 14 days of supplementation with a protein-enriched liquid (dietary sequence randomized). RESULTS: Study I: Alanine infusion increased urea synthesis rate by a factor of 10 in the control subjects, and by a factor of 5 in the CP patients (P<0.01). FHNC was 31.9+/-2.4 l/h in the control subjects and 16.5+/-2.0 l/h (P<0.05) in the CP patients. The glucagon response to alanine infusion (AUC) was reduced by 75 % in the CP patients. The reduction in FHNC paralleled the reduced glucagon response (r(2)=0.55, P<0.01). Study II: The spontaneous protein intake was 0.75+/ 0.14 g/(kg x day) and increased during the high protein period to 1.77+/-0.12 g/(kg x day). This increased alanine stimulated urea synthesis by a factor of 1.3 (P<0.05), FHNC from 13.5+/-2.6 l/h to 19.4+/-3.1 l/h (P<0.01), and the glucagon response to alanine infusion (AUC) by a factor of 1.8 (P<0.05). CONCLUSIONS: Urea synthesis rate and FHNC are markedly reduced in CP patients. This is associated with, and probably a result of, impaired glucagon secretion, and predicts a lower than normal postprandial hepatic loss of amino nitrogen. An increase in dietary protein intake increases alanine stimulated urea synthesis and FHNC by a mechanism that involves an increase in glucagon. This indicates that the low FHNC during spontaneous protein intake included an adaptation to the low protein intake, effectuated by a further decrease in glucagon secretion. PMID- 11883998 TI - Combination of recombinant human growth hormone and glutamine-enriched total parenteral nutrition to surgical patients: effects on circulating amino acids. AB - BACKGROUND & AIMS: Both recombinant human growth hormone (rhGH) and glutamine (GLN) may have beneficial anabolic actions on amino acid metabolism. The aim of this study was to evaluate the additive effects of rhGH and GLN on plasma amino acids postoperatively. METHODS: 31 females undergoing laparoscopic cholecystectomy were randomized to three groups: Group I (n=10) received 13 IU/m(2) of rhGH the morning of surgery and the following three postoperative days, together with glutamine-free TPN for the first two postoperative days. Group II (n=11) received rhGH as the first group, together with glutamine enriched (7 g GLN/m(2)/day) TPN. Group III (n=10) received glutamine-enriched TPN as the second group, but rhGH was replaced by placebo. Daily plasma amino acid concentrations and nitrogen balance were determined. RESULTS: In the GH treated groups, the plasma concentrations of several amino acids were decreased on the third postoperative day, compared to preoperatively. This was not observed in Group III. The changes were more pronounced in Group II. In Group II the negative AV-differences of amino acids tended to be attenuated, while the patients in Group III had increased negative AV-differences. The cumulative nitrogen balance was significantly improved in the GH groups, compared with Group III. CONCLUSION: The combined treatment of growth hormone and glutamine has additive effects on AV balances of amino acids postoperatively, whereas nitrogen balance is not further improved when adding glutamine to rhGH treatment. PMID- 11883999 TI - Prescribing of oral nutritional supplements in Primary Care: can guidelines supported by education improve prescribing practice? AB - BACKGROUND AND AIMS: With increasing resources being spent on nutritional supplements, this study sought to evaluate the effect of introducing guidelines on prescribing of supplements, by auditing practice, prior to, and after the implementation of guidelines. METHODS: Prescribing practice was evaluated from patient interviews, and knowledge of health professionals examined from questionnaires from 50 GP practices. Training on the use of guidelines on prescribing supplements was implemented, incorporating a Nutritional Screening Tool and practical application of high-energy dietary advice, targeting GPs and Community Nurses. RESULTS: Education to GPs and Community Nurses significantly reduced total prescribing by 15% and reduced the levels of inappropriate prescribing from 77% to 59% due to an improvement in monitoring of patients prescribed supplements. Although knowledge regarding high-energy dietary advice for nutritionally 'at risk' patients did improve as a result of the training, this was not demonstrated in practice. This lack of relevant dietary advice remained the main reason that inappropriate supplement prescriptions remained high. CONCLUSION: Education on guidelines incorporating a Nutritional Screening Tool has proved to be an effective method of achieving more appropriate prescribing of supplements, suggesting the need for ongoing training of health professionals in Primary Care. PMID- 11884000 TI - Should the food intake of patients admitted to acute hospital services be routinely supplemented? A randomized placebo controlled trial. AB - BACKGROUND: Many patients admitted to acute hospital services are underweight or harbour vitamin deficiencies. OBJECTIVES: To determine the effect on patient throughput of a policy of routine vitamin supplementation, and of early routine sipfeed supplementation in 'thin' patients (5-10% weight loss or body mass index 18-22). DESIGN: Factorial randomized placebo controlled trial of oral multivitamins from the first day of admission, and, after nutritional screening, of a nutritionally complete sipfeed from the second day in 'thin' patients. SETTING: Acute medical, surgical and orthopaedic hospital services of a London teaching hospital. PARTICIPANTS: 1561 patients admitted as emergencies were included in the vitamin study of which 549 were included in the sipfeed study. MAIN OUTCOME MEASURE: Length of hospital stay (LOS). RESULTS: Offering multivitamins to acute admissions resulted in a mean change (reduction) in LOS of -0.4 days 95% CI (-2-1.2days). The results suggest greater reductions for those discharged after 10 days: mean change=-2.3 days 95% CI (-5.7 to 1.2). Sipfeed supplementation was associated with an increased mean length of stay 2.8 days 95% CI (-0.8-6.3). 18% of acute admissions were classified undernourished on the basis of BMI, MUAC or percent weight loss combined. CONCLUSIONS: No benefit was observed for sipfeed intervention although a small benefit of less than one day is not excluded. Vitamin supplementation may have slight but economically important benefit. PMID- 11884001 TI - Colonization and bacteremia risk factors in parenteral nutrition catheterization. AB - AIMS: 1) To establish the relationship between the kind of microorganism that colonizes parenteral nutrition catheters and several risk factors related to catheterization and patient characteristics. 2) To investigate the risk factors associated to bacteremia episodes originated in these colonized catheters. METHOD: An observational, non-controlled, retrospective and cohorts study of the parenteral nutrition catheters implanted between 1988 and 1994 in our hospital. Risk factors were studied in 6 multiple-logistic regression models. RESULTS: 3632 catheters were studied. Incidences of colonization and bacteremia per 1000 days of catheterization were 17.56 and 3.93, respectively. Coagulase-negative staphyloccoci (CNS) were the most frequently isolated microorganisms. The colonization risk factors were: insertion site for all the microorganisms except fungi, catheterization time for CNS and fungi, hospitalization area, sex and age for CNS model, the existence of other infectious foci for Gram negative bacilli (GNB), S. aureus and other microorganisms, hypoalbuminemia for GNB model, and neoplasm for other microorganisms. The bacteremia risk factors were jugular insertion site, catheterization time greater than 10 days, catheter's hub colonization, and catheter colonization by gram-negative bacilli, fungi and S. aureus. CONCLUSION: Risk factors for catheter colonization vary depending on the microorganism which colonizes the catheter. PMID- 11884002 TI - A prospective comparison of the use of nasogastric and percutaneous endoscopic gastrostomy tubes for long-term enteral feeding in older people. AB - OBJECTIVE: To compare the indications for and the outcome of long-term enteral feeding by nasogastric tube (NGT) with that of percutaneous endoscopic gastrostomy (PEG) tube. DESIGN: A prospective, multicenter cohort study. SETTING: Acute geriatric units and long-term care (LTC) hospitals in Jerusalem, Israel. PARTICIPANTS: 122 chronic patients aged 65 years and older for whom long-term enteral feeding was indicated as determined by the treating physician. Patients with acute medical conditions at the time of tube placement were excluded. MEASUREMENTS: We examined the indications for enteral feeding, nutritional status, outcome and complications in all subjects. Subjects were followed for a minimum period of six months. RESULTS: Although the PEG patients were older and had a higher incidence of dementia, there was an improved survival in those patients with PEG as compared to NGT (hazard ratio (HR)=0.41; 95% confidence interval (CI) 0.22-0.76; P=0.01). Also, the patients with PEG had a lower rate of aspiration (HR=0.48; 95% CI 0.26-0.89) and self-extubation (HR=0.17; 95% CI 0.05 0.58) than those with NGT. Apart from a significant improvement in the serum albumin level at the 4-week follow-up assessment in the patients with PEG compared to those with NGT (adjusted mean 3.35 compared to 3.08; F=4.982), nutritional status was otherwise similar in both groups. CONCLUSION: In long-term enteral feeding, in a selected group of non-acute patients, the use of PEG was associated with improved survival, was better tolerated by the patient and was associated with a lower incidence of aspiration. A randomized controlled study is needed to determine whether PEG is truly superior to NGT. PMID- 11884003 TI - Treatments for Crohn's disease that minimise steroid doses are associated with a reduced risk of osteoporosis. AB - Crohn's disease is associated with an increased prevalence of osteoporosis. Corticosteroids, commonly used to control exacerbations, appear to be a major risk factor for subsequent development of osteoporosis. Exclusion diets, avoiding foods that precipitate symptoms, frequently allow control of the disease avoiding the use of corticosteroids and may thereby reduce the risk of osteoporosis. To investigate this we performed bone mineral density measurements of the proximal femur and spine in 95 patients, 31 treated predominately by corticosteroids, 33 by dietary manipulation with a low life-time corticosteroid dose and 31 by treatments other than diets but also with a low life-time corticosteroid dose. In both groups with a low life-time corticosteroid dose bone mineral density was comparable to that of age-matched normal controls, whereas bone mineral density was significantly reduced in those treated predominately by corticosteroids. We conclude that corticosteroid therapy is an independent risk factor for osteoporosis in patients with Crohn's disease and should be used as little as possible. PMID- 11884004 TI - Determination of metabolic monitor errors and precision under clinical conditions. AB - BACKGROUND & AIMS: Metabolic monitoring devices used in the critical care setting are subject to a range of conditions that may compromise their accuracy. We sought to investigate the error and precision of the Deltatrac metabolic monitor under these conditions. METHODS: A modified version of the funnel burner, described by Miodownik et al. (8), was ventilated by a mechanical ventilator. This was used to examine the performance of the Deltatrac metabolic monitor over a wide range of inspired oxygen concentrations, minute ventilation, and positive end expiratory pressure at different levels of oxygen consumption and carbon dioxide production. RESULTS: The Deltatrac measured V(O(2)) with a mean error+/ precision of 9.4%+/-19.5% (range, 9.3%+/-1.9%-72.6%+/-13.6%). The mean V(CO(2)) error+/-precision was 1.2%+/-3.1% (range-2.0%+/-1.2%-5.4%+/-3.1%). Error was significantly affected by oxygen concentration and minute ventilation but was largely independent of positive and expiratory pressure. CONCLUSIONS: The methodology of Miodownik et al. permits the expression of metabolic device errors over a wide range of simulated clinical conditions. PMID- 11884005 TI - Should a single centre for the assay of biochemical markers of nutritional status be mandatory in multicentric trials? AB - Clinical trials to identify patients at risk and to assess new therapeutic agents in the nutritional field are often single-site. The principal advantage of mounting a multicentric trial is that patient accrual is much quicker. Albumin, transthyretin, and C-reactive protein are frequently used biochemical markers of nutritional and inflammation status. However, the different techniques, reagents, and calibrators used to measure these markers introduce wide variations in values among laboratories. This study was carried out as part of a prospective multicentric study in chronic respiratory disease patients to evaluate variability and comparability of results among laboratories for these biochemical markers, and to determine whether centralization is necessary. Thirty enrolled laboratories provided their own range of reference values for those proteins a nd were then requested to process two control samples C1 and C2 blind. The results showed a broad dispersion of values for albumin and transthyretin. In 7% of laboratories, results of albumin for C2 (mean, all techniques: 39.1+/-3 g/l) were <35 g/l, the threshold value indicating a potential risk of malnutrition. When only laboratories using immunonephelemetry were considered, the results were satisfactory (CV<10% for all proteins). Given the possible incorrect classification of patients at risk, measurement should be made per site only if all participants use an immunonephelometric method. Otherwise, a centralizing assay of these biological markers should be considered. PMID- 11884006 TI - Corn might prevent Parkinson's Disease. PMID- 11884007 TI - The contribution of starvation, deconditioning and ageing to the observed alterations in peripheral skeletal muscle in chronic organ diseases. AB - Muscle weakness and early fatigue are common symptoms of chronic organ diseases, like chronic obstructive pulmonary disease (COPD), chronic heart failure (CHF) and chronic renal failure (CRF). It is becoming more and more clear that symptom intensities and exercise intolerance are related to muscle wasting and intrinsic alterations in peripheral skeletal muscle in these patient populations, while correlations with parameters of organ functioning are poor. Also, changes in muscle structure and function in COPD, CHF and CRF show much resemblance. Semi starvation, reduced physical activity and ageing are external factors possibly confounding a direct relationship between the primary organ impairments and alterations in peripheral skeletal muscle and exercise capacity. Reducing the catabolic effects of the various contributing factors might improve muscle function and health status in chronic disease. In this review, we present a systematic overview of human studies on alterations in skeletal muscle function, morphology and energy metabolism in COPD, CHF, CRF and we compare the results with comparable studies in anorexia nervosa, disuse or inactivity and ageing. Unravelling the relative contributions of these external factors to the observed alterations in the various diseases may contribute to targeted intervention strategies to improve muscle function in selected groups of patients. PMID- 11884008 TI - The reproducibility of a new dietary record routine in geriatric patients. AB - BACKGROUND AND AIM: Malnutrition in nursing home residents is an important clinical and public health problem. Knowledge is lacking about the reproducibility of dietary recording in geriatric patients. Few studies have described water intake in this age group. The aim of this study was to test the reproducibility of a 7-day dietary record routine in a clinical setting. METHODS: The dietary intake of 81 geriatric patients was recorded for two discrete periods of 7 consecutive days by the ward staff. The dietary record routine, which assessed both food and fluid intake, was based on standardized portion sizes and household measurements. RESULTS: The mean daily energy intake during the first period was 7.07 MJ and 6.84 MJ during the second period, with a mean difference of 4%. Corresponding values and the mean difference for water intake from food and beverages were 1781 g, 1702 g and 4% respectively. Age, gender, diagnosis, length of stay, diets or ADL function did not influence the results. The correlation coefficient for fluid intake between the periods was 0.84 for women and 0.72 for men. CONCLUSION: The 7-day dietary record routine seems to have a good reproducibility in assessing the intake of energy and fluids in geriatric patients. PMID- 11884009 TI - Long-term management of percutaneous endoscopic gastrostomy by a nutritional support team. AB - BACKGROUND/AIMS: Percutaneous Endoscopic Gastrostomy (PEG) has become a commonly performed procedure, to provide enteral nutrition for patients who are unable to eat. The aims of this study were to evaluate the long term efficacy, morbidity and mortality of percutaneous endoscopic gastrostomy (PEG). MATERIAL AND METHODS: We analysed 144 patients who underwent a PEG procedure. Survival curves were done with the Kaplan-Meier method. The indication was long-term enteral nutrition in patients unable to maintain adequate nutrition by mouth. RESULTS: The procedure was successful in all but one case. Mean age was 62 (18-85) years, 89 (62%) males. Seven patients recovered from their primary disease and gastrostomy tube was removed. Mean follow-up was 7.3+/-10.8 (1--66) months. Survival rates at 30 days, 1 year and 3 years following gastrostomy were 82%, 36% and 14%, respectively. Survival curves were better in females (P<0.0001). In almost all cases, patients were fed with current home-prepared food, and were ambulatory. There were no differences in survival curves according to the nutritional status. CONCLUSIONS: There were few procedure-related complications, but a high short term mortality, probably related with the underlying disease. The use of home prepared food through the gastrostomy was very well tolerated, and should be encouraged. PMID- 11884010 TI - Catheter-related infection in patients on home parenteral nutrition: results of a prospective survey. AB - BACKGROUND & AIMS: Central venous catheter (CVC) infection is the most frequent complication during home parenteral nutrition (HPN). We prospectively assessed incidence and catheter-related sepsis (CRS)-associated factors in the 42 adult patients enrolled in our HPN centre since its opening. METHODS: Age, frequency of infusions, CVC type, autonomy or nurse/family aid, underlying disease, involved infectious organism(s), hospital stay, efficacy of antibiotic-lock and other infectious complications, were studied. RESULTS: CRS occurred 39 times (3/1000 days of HPN). In 37/39 cases, it was proven by both peripheral and central blood cultures. In 56% of patients, clinical signs were discrete, delaying diagnosis. Individual factors like learning potency, underlying disease (especially chronic intestinal obstruction with bacterial overgrowth), and length of remaining colon and small intestine, were slightly associated with higher CRS incidence. Usually, one organism (S. epidermidis; 51%) was detected. A total of 14 CVC were immediately removed. In the others, antibiotic-lock was more effective in patients having tunnelled catheters (TC, 50%) than implanted devices (25%; P<0.05). Mean hospital stay was 22+/-15 days, which was influenced by 3 patients presenting associated osteomyelitis. CONCLUSIONS: CRS incidence was 3/1000 days of HPN. Clinical symptoms were often discrete, suggesting importance of rigorous survey. Individual apprenticeship and risk for higher bacterial translocation seem associated to higher CRS incidence. CVC sterilization was more frequent in patients with TC. PMID- 11884011 TI - Effects of medium-chain triglyceride in parenteral nutrition on rats undergoing gastrectomy. AB - This study was designed to investigate the effects of pre-infusion with total parenteral nutrition (TPN) using medium-chain triglyceride (MCT) versus long chain triglyceride (LCT) emulsion as fat sources on hepatic lipids, inflammatory mediators and antioxidant capacity in rats undergoing gastrectomy. Rats with internal jugular catheter, were divided into two groups and received TPN. TPN supplied 300 kcal/kg/d with 39% of the energy provided as fat. All TPN solutions were isonitrogenous and identical in nutrient composition except for the fat emulsion, which was composed of MCT/LCT (1 : 1) or LCT. After receiving TPN for 5 days, the rats underwent partial gastrectomy and were sacrificed 24 h after surgery. The results of the study demonstrated that the MCL/LCT group had lower hepatic lipids than did the LCT group. No differences in interleukin-1beta, interleukin-6 and tumor necrosis factor-alpha in peritoneal lavage fluid were observed between the two groups. Erythrocyte glutathione peroxidase activity was significantly higher in the LCT group than the MCT/LCT group, although erythrocyte superoxide dismutase activity did not differ significantly between the two groups. These results suggest that infusion with MCT/LCT before an abdominal operation did not have an effect on modulating the production of inflammatory mediators in the location of the injurious stimulus. However, pre infusion with MCT/LCT have beneficial effect in improving liver lipid metabolism and reducing oxidative stress in rats with gastrectomy. PMID- 11884012 TI - Orosomucoid: a mortality risk factor in elderly people living in the community? AB - AIM: We explored the value of variables relating to inflammation and nutrition as a prognostic factors for mortality in an elderly community-dwelling population. METHODS: We measured plasma levels of orosomucoid, C-reactive protein (CRP), albumin, and transthyretin, and the body mass index (BMI) of 245 subjects aged 65 to 95 living in the community (PAQUID study). The risk of death was analyzed two, four and six years after blood sampling by use of the Cox proportional hazards model with delayed entry. A survival curve was generated by the Kaplan-Meier method. RESULTS: Multivariate analysis--including sex, BMI, and plasma levels of albumin, transthyretin orosomucoid and CRP levels--showed that orosomucoid in the highest quartile (>0.88 g/L) was the strongest predictor of mortality two years after blood sampling [relative risk (RR)=7.4; 95% interval confidence (IC) 2.2 24.6; P<0.1]; the association remained significant four and six years after blood sampling [RR=2.5; 95%IC 1.2-5.2 and RR=1.9; 95%IC 1.0-3.4 respectively, P<0.05]. Orosomucoid levels above 1 g/L (the most accurate threshold for prediction of mortality) were associated with a strong increase in the risk of death two years [RR=12.3; 95%IC 4.3-35.0; P<0.001], four years [RR=6.9; 95%IC 3.3-14.7; P<0.001], and six years [RR=4.4; 95%IC 2.3-8.5; P<0.001) after blood sampling. CONCLUSION: These results further underline the association between systemic inflammation and mortality. They may help us to identify of high-risk subpopulations of elderly subjects so we can employ prevention strategies. PMID- 11884013 TI - Crohn's disease clinical course and severity in obese patients. AB - BACKGROUND AND AIMS: Obesity is unusual in Crohn's disease and the particularities of the disease in obese patients have not been studied. METHODS: 2065 patients were studied retrospectively. Obesity was defined by a BMI value >25.0 at disease onset and >30.0 at any time during the course of the disease. Disease characteristics, therapeutic needs, and year-by-year disease activity were determined in patients with and without obesity. RESULTS: 62 patients (3%) were obese. When compared with non-obese patients, obese patients did not show differences regarding sex, intestinal disease location, and disease behavior, but at diagnosis they were older (32 vs 28 years, P = 0.01) and a larger proportion had anoperineal disease (35 vs 24%, P = 0.03). When the 62 obese patients were paired for sex, location of disease at onset, date of birth, and date of diagnosis with 124 non-obese patients, the disease severity assessed by the importance of medical therapy and excisional surgery did not differ in the two groups but time to development of anoperineal abscess or fistula was shorter in obese patients, and obese patients were more prone to develop an active disease (OR 1.50, 95% CI 1.07-2.11) and to require hospitalization (OR 2.35, 95% CI 1.56 3.52) CONCLUSION: Obesity in Crohn's disease is associated with more frequent anoperineal complications and a more marked year-by-year disease activity, but does not alter significantly the long-term course of the disease. PMID- 11884014 TI - Feeding the gut early after digestive surgery: results of a nine-year experience. AB - BACKGROUND AND AIMS: Early enteral nutrition (EEN) after surgery should be preferred to parenteral feeding, but its clinical use is limited for concerns about possible gastrointestinal (GI) adverse effects and feeding tube-related complications. Thus we evaluated our experience focusing on safety and tolerance of early postoperative jejunal feeding and possible risk factors for gastrointestinal adverse effects. METHODS: 650 subjects treated with EEN after major digestive surgery for cancer were prospectively studied. EEN was started within 12 hours after operation via a naso-jejunal (NJ) feeding tube or a catheter-feeding jejunostomy. The rate of infusion was progressively increased to reach the nutritional goal (25 kcal/kg/day) within the 4th postoperative day. Rigorous treatment protocols for diet delivery and EEN-related GI adverse effects were applied. RESULTS: 402 patients had a jejunostomy and 248 patients a NJ tube. EEN-related GI adverse effects were observed in 194/650 patients (29.8%). In 136/194 patients, these events were successfully handled by treatment protocols. Overall the nutritional goal was achieved in 592/650 patients (91.1%). Fifty eight (8.9%) subjects had to be switched to parenteral feeding because of refractory intolerance to EEN. Intra-abdominal surgical complications and low serum albumin (<30 g/L) were the two major factors affecting tolerance. Severe jejunostomy-related complications occurred in 7/402 (1.7%) patients. EEN-related mortality was 0.1% (1/650). CONCLUSIONS: The use of the gut early after surgery is safe and well-tolerated and it should represent the first choice for nutritional support in this type of patients. PMID- 11884016 TI - A critical approach to nutritional assessment in critically ill patients. AB - BACKGROUND AND AIMS: Nutritional assessment enhances quality of nutritional care, however, its practice bemuses professionals. This prospective study aimed to identify a feasible/informative nutritional parameter in intensive care. METHODS: 44 patients (APACHE II: 23.8+/-10.1), age 58.4+/-18.6 years, were evaluated at admission: clinical data, height, weight, body mass index (BMI), tricep skinfold thickness, mid-arm circumference (MAC), mid-arm muscle circumference (MAMC), albumin, total protein and lymphocyte count. Anthropometric parameters' performance was evaluated isolated or assembled according to Blackburn and McWhirter criteria. RESULTS: Oedema increased %IW and BMI (P<0.01); muscle depletion was frequent and agreed with MAC or MAMC ranked by both criteria, P=0.02. %IW and BMI overestimated well-nourished/overweight patients, whilst arm anthropometry, mostly MAC/MAMC, shifted towards +/-50% malnutrition. Patients were not equally ranked by both criteria; McWhirter's by using percentiles clarified the distribution and showed agreement between MAC and MAMC, P=0.007, unlike Blackburn's. Mortality was higher in patients with MAC<5th percentile, P=0.003; MAC;<15th percentile was able to predict mortality and major complications. In invasive ventilated patients, severe muscle depletion was associated with mortality, P=0.05. CONCLUSION: In intensive care most nutritional assessment methods are useless; MAC is simple, feasible and if classified by percentiles may prove functional with prognostic value. PMID- 11884015 TI - Specific changes in n -6 fatty acid metabolism in patients with chronic intestinal failure. AB - BACKGROUND AND AIMS: In patients presenting severe malabsorption, essential fatty acid (EFA) deficiency can be corrected by intravenous lipids, but EFA abnormalities persist. The purpose of this study was to evaluate the role of large resection of the small bowel or malabsorption on plasma phospholipid EFA profile. METHODS: The plasma phospholipid EFA composition was measured by gas chromatography in home parenteral nutrition patients with (n=13) or without small bowel resection (n=7) and in 14 healthy subjects. RESULTS: The two groups of patients had the same nutritional status and comparable amounts of intravenous fat. In both groups, plasma fatty acid concentrations were significantly different from those observed in healthy subjects without EFA deficiency. Among them: a decrease in 18:2n -6, 22:5n -3, 22:6n -3 and an increase in 18:3n -3, 20:4n -6, 22:4n -6. Moreover, arachidonic acid to linoleic acid ratio was higher in both groups of patients, suggesting a stimulation of the elongation and desaturation of 18:2n -6. In multiple linear regression, 18:2n -6 and 20:4n -6 levels were not associated with the small bowel length, only 22:6n -3 concentration was correlated with small bowel length. CONCLUSIONS: The patients with chronic intestinal failure on home parenteral nutrition presented specific change in their EFA and an increase in the n -6 fatty acid pathway. This could be related to the severe malabsorption. PMID- 11884017 TI - Effects of oleic-rich and omega-3-rich diets on serum lipid pattern and lipid oxidation in mildly hypercholesterolemic patients. AB - AIMS: To evaluate which dietary fat elicits the best response in terms of plasma lipids, lipoproteins, and oxidative processes. METHODS: After a 4-week run-in period, 14 mildly hypercholesterolemic subjects were fed two balanced diets for 6 week periods. During the first intervention period, patients received a monounsaturated fatty acid (MUFA)-enriched diet (olive oil diet). During the second period this diet was supplemented by n-3 polyunsaturated fatty acids (PUFAs) (n-3 diet). RESULTS: After the olive oil diet, a significant decrease in total serum cholesterol (-8.54%, P<0.01), and in apolipoprotein B (Apo B) ( 10.0%, P<0.01) was observed. With the addition of n-3 fatty acids no further significant changes in serum lipid concentrations were found. However, the n-3 diet was followed by an increase in lipoperoxides in isolated native low-density lipoprotein (LDL) (67.23%, P<0.01). CONCLUSIONS: A beneficial effect on the serum lipid pattern was observed with the olive oil-enriched diet. The lack of further beneficial modifications on blood lipids and lipoproteins and the increase in the oxidative susceptibility of LDL observed after the addition of n-3 PUFA to the olive oil diet does not favor the use of this diet in hypercholesterolemic patients if it is not associated with a high intake of antioxidants. PMID- 11884018 TI - Nutrients, age and cognition. AB - Our knowledge about the influence of nutritional supplements on human cognition, especially in the elderly, rests largely on animal behavioural research and neurochemical experiments in vitro, while only a few epidemiological studies and even fewer controlled experiments in humans are reported. This is an inherent problem, due partly to the difficulty of conducting controlled nutritional experiments in humans, but may also partly be due to the gap between the research disciplines of nutritional and neurobehavioral experimental science. LEARNING OBJECTIVES: The aim of this paper is to discuss some new findings in this line of research, and to stress the importance of the need to start bridging the gap between disciplines by identifying possible human experimental models of altered cognitive function, which can elucidate the specific mechanisms of action through which nutritional supplements may enhance cognitive performance in humans in vivo. These experimental models are important because the research in this field is mostly based on epidemiological studies, which describe associations between nutrients and cognitive functions. Contrary to epidemiological studies, experimental models mimic associations between nutrients and cognition by manipulating their presumed mechanisms of action and can eventually explain the causal nature of found associations. PMID- 11884019 TI - Home artificial nutrition in chronic neurological disorders. PMID- 11884021 TI - Conflicts of interest in clinical trials. PMID- 11884023 TI - Regulation of beta2-adrenergic receptors on CD4 and CD8 positive lymphocytes by cytokines in vitro. AB - Increasing evidence points to a close relationship between the autonomic nervous system and the immune system. To further investigate mechanisms regulating beta2 adrenergic receptor (beta2R) expression in lymphocytes, the influence of cytokines on the density of beta2R on purified CD4+ and CD8+ lymphocytes was determined in vitro. beta2R were determined by means of a radioligand binding assay with (125I)iodocyanopindolol. CD4+ and CD8+ lymphocytes were incubated with catecholamines, interleukin 1beta (IL-1beta) and interleukin 2 (IL-2) for 6-72 h. The results demonstrate declining beta2R numbers on CD4+ and CD8+ lymphocytes in vitro augmented by epinephrine. IL-1beta has no effect on beta2R expression compared to medium. However, incubation with IL-2 resulted in an up-regulation of beta2R on CD8+ lymphocytes. Thus, the study demonstrates a differential regulation of beta2R on T-lymphocyte subpopulations with CD8+ lymphocytes being more susceptible to mechanisms of beta2R modulation than CD4+ lymphocytes. The findings further strengthen the concept of a close interplay between the autonomic nervous system and the immune system. PMID- 11884024 TI - Over-expression of hsp-70 inhibits bacterial lipopolysaccharide-induced production of cytokines in human monocyte-derived macrophages. AB - Cytokines released from monocytes and macrophages are major mediators of inflammation. Heat shock significantly inhibits cytokine production from these cells. To investigate whether this inhibitory effect was mediated by heat-shock proteins (HSP), we transfected human peripheral blood monocyte-derived macrophages (MDM) with HSP-70 cDNA and examined Brucella melitensis lipopolysaccharide (LPS)-induced cytokine production in transfected cells. Over expression of HSP-70 protein in the gene-transfected MDM had no effect on cytokine synthesis unless LPS was added. LPS-induced increases in production of tumour necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), IL-10 and IL-12 were significantly inhibited by the over-expression of HSP-70. However, over-expression of HSP-70 did not block LPS-induced increase in IL-6 synthesis. To further confirm these results, an antisense HSP-70 DNA oligomer was used to block HSP-70 synthesis. The inhibitory effect of HSP-70 on LPS-induced cytokine production in gene- transfected cells was completely reversed after treatment of cells with 5 microM antisense HSP-70. The same concentration of antisense HSP-70 also partially reversed heat-shock-induced inhibition of LPS-stimulated cytokine production. These results suggest that HSP-70 is involved in the regulation of LPS-induced cytokine production and that this family of proteins plays a role in mitigating adverse effects of endotoxin during infection or other pathological stresses. PMID- 11884025 TI - Implication of TNF-alpha convertase (TACE/ADAM17) in inducible nitric oxide synthase expression and inflammation in an experimental model of colitis. AB - Tumour necrosis factor-alpha (TNF-alpha) is a pro-inflammatory cytokine which is shed in its soluble form by a disintegrin and metalloproteinase (ADAM) called TNF alpha convertase (TACE; ADAM17). TNF-alpha plays a role in inflammatory bowel disease (IBD) and is involved in the expression of inducible nitric oxide synthase (iNOS) which has also been implicated in IBD. The study was designed to investigate whether colitis induced by trinitrobenzene sulphonic acid (TNBS) in rats produces an increase in TACE activity and/or expression and whether its pharmacological inhibition reduces TNF-alpha levels, iNOS expression and colonic damage in this model. TNBS (30 mg in 0.4 ml of 50% ethanol) was instilled into the colon of female Wistar rats. Saline or TACE inhibitor BB1101 (10 mg/kg/day) was administered intraperitoneally 5 days after TNBS instillation. On day 10, colons were removed and assessed for pathological score, myeloperoxidase (MPO), NO synthase (NOS), TACE enzymatic activity and protein levels, colonic TNF-alpha and NOx- levels. Instillation of TNBS caused an increase in TACE activity and expression and the release of TNF-alpha. TNBS also resulted in iNOS expression and colonic damage. BB1101 blocked TNBS-induced increase in TACE activity, TNF alpha release and iNOS expression. Concomitantly, BB1101 ameliorated TNBS-induced colonic damage and inflammation. TNBS causes TNF-alpha release by an increase in TACE activity and expression and this results in the expression of iNOS and subsequent inflammation, suggesting that TACE inhibition may prove useful as a therapeutic means in IBD. PMID- 11884026 TI - Anti-viral effect of recombinant bovine interferon gamma on bovine leukaemia virus. AB - The antiviral activity of recombinant bovine interferon gamma (rbIFN-gamma) against bovine leukaemia virus (BLV) was evaluated by an in vitro assay. rbIFN gamma was prepared using a baculovirus expression system and replication of BLV was measured by syncytium assay. Antiviral effects were observed when bovine and sheep cells were used as target cells or effector cells and treated with 0.1 unit/ml of rbIFN-gamma. Formed syncytium numbers were reduced less than 1/20 when these cells were treated with 10 units/ml of rbIFN-gamma. However, the antiviral effects on cells of heterologous species were decreased and more than 1000 units/ml of rbIFN-gamma were required to induce an anti-BLV effect on the combination of CC81 cells as target cells and Bat2Cl6 cells as effector cells, which originated from the cat and bat, respectively. When the degree of BLV production was estimated by reverse transcriptase (RT) activity, no antiviral effect of rbIFN-gamma was induced soon after the treatment, but it was evident in the cells persistently infected with BLV. These results showed that rbIFN-gamma suppresses the replication of BLV in vitro, but has effective biological activity on cells of homologous species. PMID- 11884028 TI - In situ vaccination against a non-immunogenic tumour using intratumoural injections of liposomal interleukin 2. AB - Cancers appear to escape surveillance by the immune system at least in part because they fail to induce a protective immune response. Therapeutic vaccines based on specific tumour antigens and tumour cells modified ex vivo by genetic techniques are but two strategies being used to circumvent this problem. In this report, we describe a simple, yet effective alternative in which tumour-specific responses are induced by in situ administration of a well-characterized liposomal formulation of the cytokine interleukin 2 (IL-2). Using the non-immunogenic B16 melanoma model, intratumoural injections of liposomal IL-2 L(IL2), were shown to induce a long-lived immune response specific for the injected tumour. In conjunction with subsequent removal of the primary tumours by surgery, the injections increased mean survival to 57 days from a control value of 32 days and partially protected surviving mice against re-challenge with B16. L(IL2) induced an early infiltration of inflammatory cells within the tumours which was followed several days later by an influx of CD3+ T cells. The cellular influx and a coincident decrease in tumour growth were noted in both injected tumours and a second non-injected tumour on the same animal, thereby demonstrating the systemic nature of the immune response. Intratumoural injections of soluble IL-2 at the same dose failed to induce B16-specific cellular immunity or to prolong survival of the mice. Thus, liposomal formulation of the cytokine was fundamental to successful induction of immunity by this in situ vaccination regimen. PMID- 11884027 TI - IL-6 a key cytokine in in vitro and in vivo response of Sertoli cells to external gamma irradiation. AB - Interleukin 1(IL-1) and IL-6 are cytokines involved in the response to radiation and are known for their radioprotective properties with respect to total-body irradiation. We previously showed that after gamma irradiation of Sertoli cells (SC), we observed an increase in the activity of IL-6 but not of IL-1. The aim of this study was to see whether this response is a function of the differentiation of SC, to analyse the mechanisms responsible for this induction, and to test whether this cytokine has a radioprotective role on germ cells. Unlike IL-1, a dose-dependent increase of IL-6 activity in SC following gamma irradiation at high doses was observed at all ages studied. On the other hand, radio-induction observed at low doses (<1Gy) was dose-independent. IL-6 up-regulation resulted from transcriptional activation as shown by the use of specific inhibitors. The injection of IL-1 and IL-6 in mice prior to whole-body irradiation resulted in an increased survival rate. Moreover, cytokines protected DNA from remaining cells following irradiation as shown by comet assay on germ cells. In conclusion, IL-6 seems to constitute a good marker of exposure to gamma irradiation, both at low and high doses. In addition, we showed that IL-1 and IL-6 have a radioprotective effect at testicular level. PMID- 11884029 TI - Cancer cachexia is mediated in part by the induction of IL-6-like cytokines from the spleen. AB - The development of cancer cachexia has been linked to cytokines related to interleukin6 (IL-6). We examined the kinetics of IL-6, IL-11, oncostatinM (OSM) and leukaemia inhibitory factor (LIF) induction in the splenocytes of tumour bearing mice. Using a lung carcinoma line, which grows in C57BL/6J mice, we observed that when the tumour grew and cachexia was observed, the splenocytes produced IL-6, IL-11, and OSM, but not LIF. Cytokine expression was observed within 1 week (day 3 for IL-6 and IL-11, and day 1 for OSM) of administration of tumour cells, and was observed in splenocytes without tumour metastases to the spleen. Cytokine expression preceded cachexia (determined by changes in serum triglyceride levels and decrease in epididymal fat-pad weights) development by over 1 week. Exogenous administration of IL-11 resulted in the accelerated onset of cachexia, compared to control protein treatment, but without an effect on the tumour burden. In vivo treatment with a neutralizing dose of anti-OSM antibody inhibited the triglyceride dysregulation only until the synthesis of IL-6 and IL 11 began in the spleen (day 3). Afterward, IL-6 and IL-11 induced lipid catabolism in the absence of functional OSM. We conclude from the data described above that cachexia developed due to a systemic cytokine response induced by a tumour burden, and that IL-6-like cytokines contributed independently to lipid hypercatabolism in the aetiology of cancer cachexia. PMID- 11884031 TI - Morpholino oligos: making sense of antisense? AB - Since morpholino oligos were first introduced as a means to inhibit gene function in embryos, in the Spring of 2000, they have been tested in a range of model organisms, including sea urchin, ascidian, zebrafish, frog, chick, and mouse. This review surveys the results of these studies and examines the successes and limitations of the approach for targeting maternal and zygotic gene function. The evidence so far suggests that, with careful controls, morpholinos provide a relatively simple and rapid method to study gene function. PMID- 11884032 TI - Two genetic circuits repress the Caenorhabditis elegans heterochronic gene lin-28 after translation initiation. AB - The heterochronic gene lin-28 of the nematode Caenorhabditis elegans controls the relative timing of diverse developmental events during the animal's larval stages. lin-28 is stage-specifically regulated by two genetic circuits: negatively by the 22-nt RNA lin-4 and positively by the heterochronic gene lin 14. Here, we show that lin-28 is repressed during normal development by a mechanism that acts on its mRNA after translation initiation. We provide evidence that lin-14 inhibits a negative regulation that is independent of the lin-4 RNA and involves the gene daf-12, which encodes a nuclear hormone receptor. The lin-4 independent repression does not affect the initiation of translation on the lin 28 mRNA, and like the lin-4-mediated repression, acts through the gene's 3' untranslated region. In addition, we find that lin-4 is not sufficient to cause repression of lin-28 if the lin-4-independent circuit is inhibited. Therefore, the lin-4-independent circuit likely contributes substantially to the down regulation of lin-28 that occurs during normal development. The role of lin-4 may be to initiate or potentiate the lin-4-independent circuit. We speculate that a parallel lin-4-independent regulatory mechanism regulates the expression of lin 14. PMID- 11884033 TI - A novel Dictyostelium gene encoding multiple repeats of adhesion inhibitor-like domains has effects on cell-cell and cell-substrate adhesion. AB - The Dictyostelium protein AmpA (adhesion modulation protein A) is encoded by the gene originally identified by the D11 cDNA clone. AmpA contains repeated domains homologous to a variety of proteins that influence cell adhesion. The protein accumulates during development, reaching a maximal level at the finger stage. Much of the AmpA protein is found extracellularly during development, and in culminants, AmpA is found in association with anterior-like cells. Characterization of an ampA- strain generated by gene replacement reveals a significant increase in cell-cell clumping when cells are starved in nonnutrient buffer suspensions. Developing ampA- cells are also more adhesive to the underlying substrate and are delayed in developmental progression, with the severity of the delay increasing as cells are grown in the presence of bacteria or on tissue culture dishes rather than in suspension culture. Reintroduction of the ampA gene rescues the developmental defects of ampA- cells; however, expression of additional copies of the gene in wild-type cells results in more severe developmental delays and decreased clumping in suspension culture. We propose that the AmpA protein functions as an anti-adhesive to limit cell-cell and cell-substrate adhesion during development and thus facilitates cell migration during morphogenesis. PMID- 11884035 TI - Conserved interactions with cytoskeletal but not signaling elements are an essential aspect of Drosophila WASp function. AB - Wiskott-Aldrich Syndrome proteins (WASp) serve as important regulators of cytoskeletal organization and function. These modular proteins, which are well conserved among eukaryotic species, act to promote actin filament assembly in response to cues from various signal transduction pathways. Genetic analysis has revealed a requirement for the single Drosophila homolog, Wasp (Wsp), in cell fate decisions governing specific neuronal lineages. We have used this unique developmental context to assess the contributions of established signaling and cytoskeletal partners of WASp. We present biochemical and genetic evidence that, as expected, Drosophila Wsp performs its developmental role via the Arp2/3 complex, indicating conservation of the cytoskeletal aspect of Wsp function in vivo. In contrast, we find that association with the key signaling molecules CDC42 and PIP2 is not an essential requirement, implying that activation of Wsp function in vivo depends on additional or alternative signaling pathways. PMID- 11884034 TI - Identification and characterization of a calcium channel gamma subunit expressed in differentiating neurons and myoblasts. AB - Transient elevations of intracellular calcium (calcium transients) play critical roles in many developmental processes, including differentiation. Although the factors that regulate calcium transients are not clearly defined, calcium influx may be controlled by molecules interacting with calcium channels, including channel regulatory subunits. Here, we describe the chick gamma4 regulatory subunit (CACNG4), the first such subunit to be characterized in early development. CACNG4 is expressed early in the cranial neural plate, and later in the cranial and dorsal root ganglia; importantly, the timing of this later expression correlates precisely with the onset of neuronal differentiation. CACNG4 expression is also observed in nonneuronal tissues undergoing differentiation, specifically the myotome and a subpopulation of differentiating myoblasts in the limb bud. Finally, within the distal cranial ganglia, we show that CACNG4 is expressed in placode-derived cells (prospective neurons), but also, surprisingly, in neural crest-derived cells, previously shown to form only glia in this location; contrary to these previous results, we find that neural crest cells can form neurons in the distal ganglia. Given the proposed role of CACNG4 in modulating calcium channels and its expression in differentiating cells, we suggest that CACNG4 may promote differentiation via regulation of intracellular calcium levels. PMID- 11884036 TI - Surface contraction waves (SCWs) in the Xenopus egg are required for the localization of the germ plasm and are dependent upon maternal stores of the kinesin-like protein Xklp1. AB - During the first four cell cycles in Xenopus, islands of germ plasm, initially distributed throughout the vegetal half of the egg cortex, move to the vegetal pole of the egg, fusing with each other as they do so, and form four large cytoplasmic masses. These are inherited by the vegetal cells that will enter the germ line. It has previously been shown that germ plasm islands are embedded in a cortical network of microtubules and that the microtubule motor protein Xklp1 is required for their localization to the vegetal pole [Robb, D., Heasman, J., Raats, J., and Wylie, C. (1996). Cell 87, 823-831]. Here, we show that germ plasm islands fail to localize and fuse in Xklp1-depleted eggs due to the abrogation of the global cytoplasmic movements known as surface contraction waves (SCWs). Thus, SCWs are shown to require a microtubule-based transport system for which Xklp1 is absolutely required, and the SCWs themselves represent a cortical transport system in the egg required for the correct distribution of at least one cytoplasmic determinant of future pattern. PMID- 11884037 TI - Targeted deletion of the MLC1f/3f downstream enhancer results in precocious MLC expression and mesoderm ablation. AB - The expression of skeletal muscle contractile proteins is tightly regulated during embryonic development. In the mouse, the myosin light chain (MLC) 1f/3f gene locus is not activated until E9.5, exclusively in skeletal muscle precursor cells. A potent enhancer downstream of the MLC1f/3f locus confers correct temporal and spatial activation of linked reporter gene in transgenic mouse embryos. To examine roles of the MLC downstream enhancer (MLCE) in its native context of the MLC1f/3f gene locus, we eliminated a 1.5-kb DNA segment containing the enhancer from the mouse genome by targeted deletion, leaving no exogenous sequences at the deletion site. Mouse embryos homozygous for the MLCE deletion were smaller and developmentally delayed, formed no mesoderm by E7.5, and were resorbed almost completely at E8.5. In situ hybridization and RT-PCR analyses of affected mutant embryos at E7.5 revealed ectopic MLC transcripts, whose products would be predicted to interfere with a variety of nonmuscle cell functions determining differentiation of mesoderm. These results suggest that the MLC downstream enhancer and its flanking sequences include negative regulatory elements which block precocious activation of MLC expression in mesodermal precursors during a critical window of development, as well as positive elements which subsequently permit tissue-restricted MLC transcription in differentiating skeletal muscles. PMID- 11884038 TI - Mouse parthenogenetic embryos with monoallelic H19 expression can develop to day 17.5 of gestation. AB - In mammals, both maternal and paternal genomes are required for a fetus to develop normally to term. This requirement is due to the epigenetic modification of genomes during gametogenesis, which leads to an unequivalent expression of imprinted genes between parental alleles. Parthenogenetic mouse embryos that contain genomes from nongrowing (ng) and fully grown (fg) oocytes can develop into 13.5-day-old fetuses, in which paternally and maternally expressed imprinted genes are expressed and repressed, respectively, from the ng oocyte allele. The H19 gene, however, is biallelically expressed with the silent status Igf2 in such parthenotes. In this study, we examined whether the regulation of H19 monoallelic expression enhances the survival of parthenogenetic embryos. The results clearly show that the ng(H19-KO)/fg(wt) parthenogenetic embryos carrying the ng-oocyte genome that had been deleted by the H19 transcription unit successfully developed as live fetuses for 17.5 gestation days. Control experiments revealed that this unique phenomenon occurs irrespective of the genetic background effect. Quantitative gene expression analysis showed that day 12.5 ng(H19-KO)/fg(wt) parthenogenetic fetuses expressed Igf2 and H19 genes at <2 and 82% of the levels in the controls. Histological analysis demonstrated that the placenta of ng(H19 KO)/fg(wt) parthenotes was afflicted with atrophia with severe necrosis and other anomalies. The present results suggest that the cessation of H19 gene expression from the ng-allele causes extended development of the fetus and that functional defects in the placenta could be fatal for the ontogeny. PMID- 11884039 TI - AlphaIIb integrin expression during development of the murine hemopoietic system. AB - Integrin alphaIIb is a cell adhesion molecule expressed in association with beta3 by cells of the megakaryocytic lineage, from committed progenitors to platelets. While it is clear that lymphohemopoietic cells differentiating along other lineages do not express this molecule, it has been questioned whether mammalian hemopoietic stem cells (HSC) and various progenitor cells express it. In this study, we detected alphaIIb expression in midgestation embryo in sites of HSC generation, such as the yolk sac blood islands and the hemopoietic clusters lining the walls of the major arteries, and in sites of HSC migration, such as the fetal liver. Since c-Kit, which plays an essential role in the early stages of hemopoiesis, is expressed by HSC, we studied the expression of the alphaIIb antigen in the c-Kit-positive population from fetal liver and adult bone marrow differentiating in vitro and in vivo into erythromyeloid and lymphocyte lineages. Erythroid and myeloid progenitor activities were found in vitro in the c Kit(+)alphaIIb(+) cell populations from both origins. On the other hand, a T cell developmental potential has never been considered for c-Kit(+)alphaIIb(+) progenitors, except in the avian model. Using organ cultures of embryonic thymus followed by grafting into athymic nude recipients, we demonstrate herein that populations from murine fetal liver and adult bone marrow contain T lymphocyte progenitors. Migration and maturation of T cells occurred, as shown by the development of both CD4(+)CD8- and CD4-CD8(+) peripheral T cells. Multilineage differentiation, including the B lymphoid lineage, of c-Kit(+)alphaIIb(+) progenitor cells was also shown in vivo in an assay using lethally irradiated congenic recipients. Taken together, these data demonstrate that murine c Kit(+)alphaIIb(+) progenitor cells have several lineage potentialities since erythroid, myeloid, and lymphoid lineages can be generated. PMID- 11884040 TI - Direct flight muscles in Drosophila develop from cells with characteristics of founders and depend on DWnt-2 for their correct patterning. AB - The direct flight muscles (DFMs) of Drosophila allow for the fine control of wing position necessary for flight. In DWnt-2 mutant flies, certain DFMs are either missing or fail to attach to the correct epithelial sites. Using a temperature sensitive allele, we show that DWnt-2 activity is required only during pupation for correct DFM patterning. DWnt-2 is expressed in the epithelium of the wing hinge primordium during pupation. This expression is in the vicinity of the developing DFMs, as revealed by expression of the muscle founder cell-specific gene dumbfounded in DFM precursors. The observation that a gene necessary for embryonic founder cell function is expressed in the DFM precursors suggests that these cells may have a similar founder cell role. Although the expression pattern of DWnt-2 suggests that it could influence epithelial cells to differentiate into attachment sites for muscle, the expression of stripe, a transcription factor necessary for epithelial cells to adopt an attachment cell fate, is unaltered in the mutant. Ectopic expression of DWnt-2 in the wing hinge during pupation can also create defects in muscle patterning without alterations in stripe expression. We conclude that DWnt-2 promotes the correct patterning of DFMs through a mechanism that is independent of the attachment site differentiation initiated by stripe. PMID- 11884041 TI - SNARE complex assembly is required for human sperm acrosome reaction. AB - Exocytosis of the acrosome (the acrosome reaction) is a terminal morphological alteration that sperm must undergo prior to penetration of the extracellular coat of the egg. Ca(2+) is an essential mediator of this regulated secretory event. Aided by a streptolysin-O permeabilization protocol developed in our laboratory, we have previously demonstrated requirements for Rab3A, NSF, and synaptotagmin VI in the human sperm acrosome reaction. Interestingly, Rab3A elicits an exocytotic response of comparable magnitude to that of Ca(2+). Here, we report a direct role for the SNARE complex in the acrosome reaction. First, the presence of SNARE proteins is demonstrated by Western blot. Second, the Ca(2+)-triggered acrosome reaction is inhibited by botulinum neurotoxins BoNT/A, -E, -C, and -F. Third, antibody inhibition studies show a requirement for SNAP-25, SNAP-23, syntaxins 1A, 1B, 4, and 6, and VAMP 2. Fourth, addition of bacterially expressed SNAP-25 and SNAP-23 abolishes exocytosis. Acrosome reaction elicited by Rab3-GTP is also inhibited by BoNT/A, -C, and -F. Taken together, these results demonstrate a requirement for members of all SNARE protein families in the Ca(2+)- and Rab3A triggered acrosome reaction. Furthermore, they indicate that the onset of sperm exocytosis relies on the functional assembly of SNARE complexes. PMID- 11884042 TI - Five-node biopsy of the axilla: an alternative to axillary dissection of levels I II in operable breast cancer. AB - BACKGROUND: Axillary clearance of patients with early breast cancer is accompanied by a high risk of arm morbidity. Less invasive ways to establish the axillary nodal status are therefore of interest, especially in women with low risk of nodal metastases. METHODS: Four hundred and fifteen breast cancer patients (clinical stage T(0-3) N(0-1) M(0)) were operated in the axilla with a five-node biopsy followed in the same operation by a further dissection of levels I-II of the axilla in order to evaluate the accuracy of the five-node node biopsy compared with level I-II dissection. RESULTS: In all patients the sensitivity of the five-node biopsy was 97.3% with a negative predictive value of 98.5% and a negative likelihood ratio of 0.027. Among cases detected by screening (n=204) and those clinically detected (n=197) the sensitivity of the five-node biopsy was 95.8% and 97.9% respectively, with negative predictive values of 98.7% and 98.0% and negative likelihood ratios of 0.042 and 0.021 respectively. CONCLUSION: Five node biopsy of the axilla has good accuracy for correctly staging the axilla in both clinically and screening-detected cases. Five-node biopsy is an alternative to axillary clearance and sentinel node biopsy in patients with operable breast cancer. PMID- 11884043 TI - An immunohistochemical study of p21 and p53 expression in primary node-positive breast carcinoma. AB - AIMS: p21, an inhibitor of cyclin-dependent kinase, is involved in the p53 pathway of growth control. Its expression has been linked to cellular differentiation. It has been implicated in p53-mediated growth arrest following DNA damage and in terminally differentiated cells. This study analysed p21 and p53 expression, in a series of node-positive patients with breast carcinoma and examined histopathological parameters of the tumour and the prognostic implications of p21 and p53 expression. METHODS: One hundred and five consecutive patients with node-positive disease and at least 3 years follow-up were identified. Sections were stained for p53 and p21 using monoclonal antibodies. Results were expressed as percentage positive cells, and over 20% considered positive for p53 and over 10% considered for p21. RESULTS: p21 was overexpressed (>10% of cells positive) in 65% of patients and p53 was overexpressed (>20% of cells positive in 68%. The mean (SEM) level of p21 staining was 5.7(0.8)% and was 54.9(4.0)% for p53. There was no correlation between p21 and p53 expression (r=0.071 P=0.5). There were no significant differences in demographic criteria between patients that were p21 positive or negative and p53 positive or negative. There were no significant differences in tumour type, grade or stage between the groups. p21 expression did not have prognostic significance; however, p53 positivity was associated with a worse prognosis, which remained when controlled for stage. CONCLUSIONS: This study demonstrated p21 overexpression in 65% of patients with node-positive breast carcinoma. Levels did not correlate with p53 status and unlike p53 failed to have prognostic significance. PMID- 11884044 TI - The role of Tc99m-sestamibi scintimammography in combination with the triple assessment of primary breast cancer. AB - AIM: The aim of this study was to evaluate if Tc99m-sestamibi scintimammography in addition to the triple assessment consisting of clinical examination, mammography, breast ultrasonography and fine needle aspiration cytology (FNA) enhances the diagnosis of breast cancer and helps in avoiding unnecessary operative biopsies. METHODS: Pre-operational scintimammography was performed within 2 weeks of operation to 46 consecutive patients with abnormal findings in clinical breast examination, mammography or ultrasonography. Three patients had abnormalities in both breasts. Histological diagnosis was obtained in all 49 cases. RESULTS: The histological diagnosis was benign in 18 (37%) cases and malignant in 31 (63%) cases. The overall sensitivity of scintimammography was 77% and the specificity was 61%. The sensitivity of scintimammography was 95% in invasive ductal carcinoma, 50% in invasive lobular carcinoma and 25% in ductal carcinoma in situ. Scintimammography showed 100% sensitivity in cases with invasive carcinoma, with highly suspicious findings for malignancy in the other examinations. The sensitivity was 63% in cases with indeterminate or contradictory findings in mammography, ultrasonography and FNA. CONCLUSIONS: Adding scintimammography to the triple assessment does not seem to be helpful in the diagnosis of breast abnormalities because of low sensitivity in malignant cases with a challenging diagnosis by mammography, ultrasonography and FNA, and because of low overall specificity. PMID- 11884045 TI - Impact of splenectomy on the early outcome after oesophagectomy for squamous cell carcinoma of the oesophagus. AB - AIM: Operative procedures for oesophageal malignancies are becoming more extensive and may result in fatal complications. Splenectomy compromises the immune system and can lead to increased susceptibility to infections. The aim of the present study was to report the early outcome of patients who underwent oesophagectomy and simultaneous splenectomy due to oesophageal squamous cell carcinoma (SCC). METHODS: Pre-operative risks and post-operative morbidity and mortality in 135 patients who had undergone extensive oesophagectomy without simultaneous splenectomy for SCC of the thoracic oesophagus were compared with those of 14 patients who had undergone oesophagectomy associated with splenectomy. RESULTS: Post-operative pneumonia, intra-abdominal abscess, post operative sepsis and anastonotic leakage were significantly increased when splenectomy was added to the original operation. The incidence of in-hospital death was significantly higher among splenectomized than non-splenectomized patients (35.7% vs 8.1%, P<0.01). Pulmonary complications and leakage were the main causes of death. Multivariate analysis recognized splenectomy as an independent prognostic factor for in-hospital death following transthoracic oesophagectomy for SCC. CONCLUSION: The addition of splenectomy to transthoracic oesophagectomy for oesophageal carcinoma can be a fatal combination. Preservation of the spleen should be the primary intention. In circumstances that necessitate splenectomy precautions should be taken to prevent post-operative infectious complications. PMID- 11884046 TI - Fas ligand is up-regulated during the colorectal adenoma-carcinoma sequence. AB - AIMS: Fas ligand (FasL) expression by cancer cells may mediate tumour immune privilege. The purpose of the present study was to investigate the timing and significance of FasL expression during the colorectal adenoma-carcinoma sequence. METHODS: FasL expression was studied by immunohistochemistry in 170 formalin fixed tissue sections representing the entire colorectal adenoma-carcinoma sequence. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to search for FasL mRNA. Analysis of survival was performed in patients with carcinomas. RESULTS: A significant positive linear correlation was found between FasL expression and tumour progression throughout the colorectal adenoma carcinoma sequence (r(s)=0.677 P<0.001). A pattern of high FasL expression was detected in 19% of high grade adenomas, 40% of stage I-II, 67% of stage III and 70% of stage IV carcinomas. No significant differences were observed between FasL expression in the primary tumours and that in the corresponding liver metastases. The specificity of FasL expression was confirmed at RT-PCR. For stage I-II carcinomas, the 5 year survival was 90% in patients without, or with moderate, tumoural FasL expression compared with 60% in those with high tumoural FasL expression (P<0.05). CONCLUSIONS: Our findings suggest that FasL expression may be involved in the development of colorectal cancer and its progression. PMID- 11884047 TI - Local recurrence after mesorectal excision for rectal cancer. AB - AIMS: Controversy still exists about the optimal surgical treatment of rectal cancer. The main purpose of the present study was to compare local recurrence (LR) rates after mesorectal excision (ME) and conventional surgery (CS) technique. METHODS: All rectal cancer patients from a defined catchment area were included. Outcome after ME in the period 1993-1999 (n=161) was compared with the outcome after CS (n=217) in the period 1983-1992. Partial ME (PME) was the routine in upper, and total ME the routine in mid- and low rectal cancer. The follow-up programmes were identical, and the median observation times very similar (37 and 38 months) in the two periods. Five-year actuarial LR rate and survival were estimated using the Kaplan-Meier method, and adjustment for prognostic factors was performed with Cox regression analysis. RESULTS: Total LR rate after R0 resection was 7.7% crude and 9% 5 year actuarial in the ME period, as compared with 16.0% crude and 24% actuarial in the CS period (P=0.02). Cox regression analyses confirmed these differences with a hazard ratio of 0.40 for ME vs CS (P=0.02). Isolated LR rate was 2% after ME and 8% after CS. Five-year actuarial total LR rate after rectal resection with curative intent was 11% after ME and 27% after CS (P<0.01). Actuarial total LR rate after PME was 6%, and none of these patients developed isolated LR. CONCLUSION: Standardization of surgical technique and application of ME resulted in a significant reduction of LRs. LR rate was low after PME, indicating that this procedure is adequate in upper rectal cancer. PMID- 11884048 TI - Liver resection for metastatic non-colorectal non-neuroendocrine hepatic neoplasms. AB - AIMS: Major liver surgery can be performed safely and hepatic resection for metastatic disease is increasingly carried out. However, the role of liver resection for hepatic metastases from non-colorectal, non-neuroendocrine (NCNN) cancers is unknown. Our aim was to evaluate our experience from hepatectomies for NCNN metastases. A retrospective study of 170 patients with liver resection performed the last 8 years was performed in two liver units in affiliated university hospitals. METHODS: Eighteen patients underwent liver resection for NCNN tumours. Origins included kidney (n=6), breast (n=4), gastric tumours (n=4), intestinal leiomyosarcoma (n=2) and malignant melanoma and in one patient a metastatic papillary of unknown origin was found. Eleven patients underwent a hepatic lobectomy and seven had local resections. Ten hepatectomies were performed at the same time with the primary tumour resection (synchronous resections) with five of those in an en bloc fashion with the primary tumour. RESULTS: There were no post-operative deaths and the peri-operative morbidity was minimal. During a median follow-up time of 3.2 years, 14 patients are alive with one of them having developed pulmonary metastases. CONCLUSION: In carefully selected patients with NCNN liver metastasis, liver resection can prolong survival as well and improve quality of life. PMID- 11884050 TI - Diagnosis and treatment of isolated neck metastases of adenocarcinomas. AB - AIMS: Cervical metastases of adenocarcinoma or undifferentiated large cell carcinoma (ULCC) (non-squamous cell carcinoma) of unknown primary origin are rare and often accompanied by distant metastases at multiple sites in the body. Nevertheless, in the past decades, several patients have presented in our clinic with isolated neck metastases of this type of malignancy. The aim of our study is to evaluate the clinical behaviour of these cases and to define the role of surgery and radiotherapy. METHODS: Over the past 24 years, we selected 15 out of 270 patients (6%) with isolated cervical lymph node metastases of adenocarcinoma (six) or ULCC (nine) of unknown primary origin. Diagnosis was made either by histology or by fine needle aspiration cytology. Treatment consisted of (selective) neck dissection and/or radiotherapy. RESULTS: The clinical presentation of isolated cervical metastases of adenocarcinoma compared with ULCC is equivalent, with an overall median survival time of 25 months (confidence interval 21--29 months). Combined therapy was correlated with an increased and persistent regional control and was associated with longer duration of survival. CONCLUSIONS: Patients with isolated cervical neck node metastases of adenocarcinoma or ULCC of unknown primary origin are rare and the diagnostic process to identify this subgroup requires a systemic work-up. In selected cases treatment should concentrate on (selective) neck dissection combined with radiotherapy to achieve a prolonged survival. PMID- 11884049 TI - Immunohistochemical prognostic indicators of gastrointestinal carcinoid tumours. AB - AIMS: The aim of this study was to determine whether expression of the oncoproteins p21, p53, E-cadherin (EC), cyclin D1, bcl-2 and Rb and the proliferation marker Ki-67 is predictive of malignant behaviour in gastrointestinal carcinoid tumours. METHODS: Immunohistochemical (IHC) staining was performed on carcinoid tumours from 41 patients (31 rectal, eight gastrointestinal, two appendiceal lesions). The six tumours that had invaded deeply into the muscularis propria or beyond, had metastasized to regional lymph nodes or had metastasized to a distant site were classified as the malignant group, and the other 35 tumours formed the benign group. IHC expression was compared between the two groups, and the prognostic value of each marker was assessed. RESULTS: Of the six tumours in the malignant group, 66.7% were p21 positive, 0% were p53 positive, 33.3% were EC positive, 100% were cyclin D1 positive, 33.3% were Rb positive, 16.7% were bcl-2 positive and 50% were Ki-67 positive. Of the 35 tumours in the benign group, 17.1% were p21 positive, 0% were p53 positive, 100% were EC positive, 94.3% were cyclin D1 positive, 8.6% were Rb positive, 17.1% were bcl-2 positive and 0% were Ki-67 positive. CONCLUSIONS: These data show that p53, cyclin D1, Rb, bcl-2 and Ki-67 staining does not correlate with malignant behaviour but that overexpression of p21 (P=0.02) and reduced staining of EC (P=0.005) do correlate with malignant behaviour. These two parameters may therefore be useful as prognostic indicators for gastrointestinal carcinoid tumours. PMID- 11884051 TI - Prognostic factors in soft tissue sarcomas: a study of 395 patients. AB - AIMS: The aim of this study was to report prognostic factors, end-points of local recurrence, distant recurrence, post-metastasis survival, and overall survival in a cohort of patients with soft tissue sarcomas. METHODS: We analysed a database of 395 patients affected by primary soft tissue sarcomas of various primary sites, treated and followed up at the Centro di Riferimento Oncologico, Aviano, Italy from January 1985 to January 1997. RESULTS: Grade, size, stage, surgical margins, distant metastasis, age, sex, performance status, and haemoglobin value were significant for overall survival. Histology, grade, stage, and surgical margins were significant for local recurrence. Grade, size, and stage, were significant for distant recurrence; and surgical margin was significant variable for post-metastasis survival. CONCLUSIONS: Grade, size, and TNM stage (UICC/AJCC) have stronger prognostic significance for overall survival and distant recurrence than for local relapse. Positive surgical margins are the main predictors for local relapse. Age was the most consistent adverse independent prognostic factor for survival. PMID- 11884052 TI - Combined regional and systemic chemotherapy for advanced and inoperable non-small cell lung cancer. AB - AIMS: The objective was to establish the feasibility and toxicity of regional chemotherapy using an isolated thoracic perfusion (ITP) technique plus low dose systemic chemotherapy as induction chemotherapy followed by surgery in advanced non-small cell lung cancer (NSCLC). METHODS: twenty-two chemotherapy-naive patients with NSCLC (median age of 57 years, stage III-IV disease with metastases only in the thoracic region, Karnofsky index >60), received two cycles of regional plus systemic chemotherapy with a treatment-free interval of 4 weeks. The cytostatic regimen consisted of 10 mg/m(2) mitomycin, 25 mg/m(2) navelbine and 30 mg/m(2) cisplatin during ITP followed by low-dose systemic chemotherapy with 250 mg/m(2) 5-fluorouracil and 20 mg/m(2) cisplatin given as a continuous infusion on day 1-4. Patients were re-evaluated for response and surgery was carried out if possible. RESULTS: All 22 patients could be assessed for toxicity, response and survival. There were 19/22 remissions corresponding to a regression rate of 86.4%; 16/22 patients could be resected. This corresponded to a resectability rate of 72.7% (13 complete resections R0, 1 R1, 2 R2). Side-effects were transient and acceptable with no treatment- or surgery-related deaths. Median survival has not been reached after an observation time of 15 months. The estimated 1-year survival rate was 67.3%. CONCLUSIONS: Regional chemotherapy using an ITP application form is highly effective in advanced NSCLC stage III-IV leading to a high rate of resectability with an encouraging survival outcome. PMID- 11884053 TI - Intraperitoneal mitoxantrone: a feasibility and pharmacokinetic study. AB - Fractionated doses have been advocated to prevent chemoperitonitis after intraperitoneal infusion of mitoxantrone. Patients with peritoneal carcinomatosis of various origin underwent surgery, including intestinal resections, with minimal residual disease. Peritoneal mitoxantrone in 1000 ml/m(2) saline was planned on the first post-operative day in groups of four patients (5 mg/m(2) for 3 and 5 days, 7.5 mg/m(2) for 3 and 4 days, 10 mg/m(2) for 2-4 days, if possible). Due to dose-limiting myelosuppression, only one and three patients received the 7.5-mg 4-day and 10-mg 3-day regimens, respectively. A total of 20 patients were consequently treated. Neither major complications nor severe pain were observed. Pharmacokinetics were completed on the 1st day in five 5-mg and five 10-mg patients, on the 5th day in three 5-mg patients, and on the 3rd day in one 10-mg patient. On the 1st day, mean peritoneal peak concentrations of mitoxantrone resulted 1.45 +/-0.56 (range 0.48-1.9) and 1.9+/-0.85 (range 1.27 3.13) microg/ml in the 5-mg and 10-mg patients, respectively. Mean dialysate/plasma exposure (AUC) ratio was 115. Even in patients with sutures, early post-operative fractionated intraperitoneal mitoxantrone appears feasible and safe, with a high local advantage, for up to 5 days of treatment and a maximum tolerated total dose of 20-25 mg/m(2). PMID- 11884055 TI - Modifications to the double-staple technique for oesophago-jejunal anastomosis. AB - A new instrument is described that makes a small modification to the anvil of an Ethicon circular stapler, allowing it to be more easily withdrawn through a cross stapled oesophagus. This allows the double-stapling technique for oesophago jejunal anastomoses to be completed safely through the hiatus, even many centimetres inside the chest. The technique has been used on 17 anastomoses without difficulty, clinical leak or subsequent stricture. Several of these patients would have needed a thoracotomy to complete the anastomosis had it not been for this instrument. PMID- 11884054 TI - Intraoperative radiotherapy: current thinking. AB - Intraoperative radiotherapy (IORT) refers to the delivery of irradiation at surgery. A large single dose of irradiation is delivered to a surgically defined area, while uninvolved and dose-limiting tissues are displaced, the final goal of IORT being enhanced locoregional tumour control. IORT is used in most modern protocol studies as a boost radiation component of multidisciplinary treatment approaches. More recently, high activity radiation sources or mobile operating room treatment machines are used to facilitate the IORT procedure. Clinical experiences have shown that IORT may improve local control and disease-free survival, especially when used in adjuvant setting, combined with external beam irradiation in some neoplasms such as cancer of the stomach, pancreas, colorectum, and soft tissue sarcoma. PMID- 11884056 TI - Dermatitis artefacta of the breast: a series of case reports. AB - Dermatitis artefacta of the breast is a common disease process with a psychological basis. Psychiatric assessment is a useful adjunct in the management of these patients who at initial presentation seem to have a strictly surgical dermatological problem. PMID- 11884057 TI - Peritoneal benign cystic mesothelioma: a case report and review of the literature. AB - Peritoneal benign cystic mesothelioma is a rare tumour of unknown aetiology. It usually presents with mild abdominal pain and a solid tumour on physical examination. The differential diagnosis with solid abdominal tumours is difficult. Computed tomography, magnetic resonance imaging as well as aspiration cytology are useful in suggesting the pre-operative diagnosis. We present one case report and discuss this entity. PMID- 11884058 TI - Primary anorectal lymphoma presenting as a perianal abscess in an HIV-positive male. PMID- 11884059 TI - Primary primitive neuroectodermal tumour of the small bowel mesentery: case report. PMID- 11884060 TI - Recurrence in the axilla after sentinel lymph node biopsy for breast cancer. PMID- 11884061 TI - Androgen and estrogen metabolism during sex differentiation in mono-sex populations of the Nile tilapia, Oreochromis niloticus. AB - Androgen and estrogen metabolism were examined in the period of steroid sensitivity during sex differentiation in mono-sex populations of Oreochromis niloticus. Fry (XX, XY, and YY genotypes) were maintained at 28 degrees and were sampled at 8, 10, 11, and 13 days postfertilization. Subsamples (n = 2-4) of pooled fry from each maternally distinct family were homogenized and incubated with either [(3)H]androstenedione or [(3)H]estradiol. Metabolites present in organic extracts were identified by thin-layer chromatography, microchemical reactions, and recrystallization to constant specific activity. Androstenedione was metabolized into at least seven readily identifiable compounds by all genotypes. In the XY genotype, 5beta-androstane-3alpha,17beta-diol synthesis decreased rapidly from 8 to 13 days postfertilization, with a concomitant increase in testosterone synthesis. Testosterone synthesis did not increase in the XX genotype. Testosterone synthesis in the YY genotype was intermediate to that of the XY and XX genotypes. Estrogens were not synthesized by any genotype. We hypothesize that 5beta-reduction (or further hydroxylation) is a mechanism important in regulating testosterone production and subsequent sex differentiation. Results of incubations with estradiol show an age-dependent increase in metabolism which did not vary among genotypes. Metabolites synthesized included estrone and up to five unidentified compounds. PMID- 11884062 TI - The effect of water temperature on the GABAergic and reproductive systems in female and male goldfish (Carassius auratus). AB - This study investigated the effect of water temperature on the synthesis of the amino acid neurotransmitter gamma-aminobutyric acid (GABA). In goldfish, GABA stimulates the release of pituitary gonadotropin-II (GTH-II), which regulates gonadal function. Fish were maintained in water of 11, 18, or 24 degrees. In the female and male goldfish, GABA synthesis rates estimated following inhibition of GABA catabolism by gamma-vinyl GABA (GVG) in both the telencephalon (TEL) and the hypothalamus (HYP) were increased in fish held at 24 degrees compared to those at either 11 or 18 degrees (P < 0.05). Additionally, GABA synthesis rates in the pituitary increased in a temperature-dependent manner. Glutamate is the precursor for GABA synthesis; however, no consistent pattern was seen between glutamate and GABA synthesis rates, indicating that glutamate is not a limiting factor in GABA synthesis. Both water temperature and GVG administration increased serum GTH-II levels in female goldfish. However, in male goldfish water temperature had no significant effect on serum GTH-II levels, and GVG injection increased serum GTH II levels only in fish maintained at 24 degrees. The effects of temperature on the levels of mRNA expression of the GABA-synthesizing enzymes glutamate decarboxylase 65 (GAD(65)) and GAD(67) were measured by semiquantitative PCR. In the TEL and HYP of female goldfish, GAD(65) was not affected, whereas temperature change from 11 to 18 degrees increased (P < 0.05) GAD(67) mRNA levels. These results demonstrate that central GABAergic systems in the goldfish are temperature sensitive. PMID- 11884063 TI - Hematological effects of high dose of cortisol on the carp (Cyprinus carpio L.): cortisol effect on the carp blood. AB - The level of circulating cortisol and peripheral blood parameters were determined in carp age 2 years (K(2)) 24, 72, and 216 h after a single intraperitoneal injection of a high dose (200 mg x kg(-1) body wt) of hydrocortisone. The most striking effect of cortisol was manifest as a significant change in the percentage composition of leukocytes, whose number per unit volume of blood remained relatively constant. A profound lymphopenia and eosinopenia were compensated for in the general balance by an increased number of circulating promyelocytes and myelocytes as well as metamyelocytes and mature polymorphonuclear neutrophilic granulocytes. The results and their possible reasons are discussed on the background of literature data. PMID- 11884064 TI - Seasonal variations of vitelline envelope proteins, vitellogenin, and sex steroids in male and female eelpout (Zoarces viviparus). AB - The seasonal variations of vitelline envelope proteins, vitellogenin (VTG), and reproductive steroids were investigated in feral male and female eelpout, Zoarces viviparus. 17beta-Estradiol was present in both sexes with a peak in prespawning fish of 2.6 ng/ml in males and 2.7 ng/ml in females. 11-Ketotestosterone peaked in June at 4.2 and 0.47 ng/ml in males and females, respectively. A surge of testosterone was seen in both sexes in August, just prior to spawning. All steroid levels were low during early pregnancy. The vitelline envelope of the eelpout is composed of two major and one minor protein with molecular weights of 50, 55, and 44 kDa, respectively. An antiserum raised against solubilized vitelline envelope from turbot (Scophthalmus maximus) cross-reacted strongly with the 50-kDa protein from the isolated vitelline envelope and a similar-sized protein in female plasma and plasma from estrogenized males. Interestingly, the 50-kDa protein was also present at low levels in males as demonstrated by ELISA and Western blotting. In males, the 50-kDa protein did not follow the seasonal changes in 17beta-estradiol, but instead showed an almost perfect negative correlation with water temperature. VTG was present in female plasma as shown by Western blotting, but VTG was not detectable in male plasma despite relatively high endogenous estrogen levels. This suggests that the VTG induction by estradiol may be modulated by other factors in the eelpout. PMID- 11884065 TI - Variation within and between birds in corticosterone responses of great tits (Parus major). AB - The present study investigated inter- and intraindividual variation of the stress response (in terms of plasma levels of corticosterone) to handling in birds. Individual captive great tits (Parus major) were exposed to a standardised capture and handling protocol three times at about 2-week intervals. Mean plasma corticosterone levels were low (<5 ng/ml) when the birds were first sampled and increased in all birds 10 min after handling (mean levels on each sampling occasion 14 to 22 ng/ml). Levels in some birds then continued to increase, whereas in other birds levels remained relatively constant or had declined 30 min after handling began. Corticosterone responses were measured three times in each bird so that variation in plasma corticosterone levels could be defined. The area under the corticosterone response curve was defined as the integrated corticosterone response and was calculated for each response curve. Variation between birds was quantified by comparing mean values of corticosterone levels and integrated corticosterone responses between birds, then calculating a coefficient of variation for the mean of the individual bird means for each corticosterone parameter. Variation within birds was quantified by calculating the mean of the coefficients of variation for each bird for each corticosterone parameter. Variation in corticosterone levels in the first blood samples at 0 min (coefficient of variation (CV) 84.0%) was greater than variation in peak levels at 10 min (CV 35.1%) and in integrated corticosterone responses (CV 31.9%), indicating that corticosterone responses were more consistent between birds than were basal corticosterone levels. This study has demonstrated that corticosterone responses to a stressor tend to be repeatable in individual great tits, and has provided a method for quantifying variation in corticosterone responses for comparison with other birds in the future. PMID- 11884066 TI - Functional analysis of natriuretic peptide receptors in the bladder of the toad, Bufo marinus. AB - This study aimed to localize and characterize natriuretic peptide binding sites in the urinary bladder of Bufo marinus and to then examine the effect of natriuretic peptides on the bladder vascular tone and water reabsorption in isolated perfused bladder preparations. Specific (125)I-rat atrial natriuretic peptide ((125)I-rANP) binding sites were present on blood vessels, muscle, and epithelium. In tissue sections and/or isolated membranes, the binding was completely displaced by frog ANP, rat ANP, and porcine C-type natriuretic peptide (CNP; membranes only). However, a reduction in binding was observed after incubation with (125)I-rANP and 1 microM of the natriuretic peptide receptor-C (NPR-C) ligand C-ANF, but residual binding remained suggesting the presence of two distinct binding sites. Electrophoresis of bladder membranes cross-linked to (125)I-rANP identified two bands at approximately 70 and 140 kDa that correspond to the monomeric mass of NPR-C and the guanylate cyclase receptors, respectively. Furthermore, the presence of natriuretic peptide receptor-A and NPR-C mRNA in the bladder was demonstrated with reverse transcription--polymerase chain reaction. In addition, rat ANP, frog ANP, and porcine CNP stimulated a significant increase in cGMP generation in bladder membrane preparations, which indicated the presence of guanylate cyclase-linked receptors. In perfused bladder preparations, arginine vasotocin increased perfusion pressure and water permeability. The infusion of frog ANP or porcine CNP failed to alter perfusion pressure or water reabsorption in the presence or absence of arginine vasotocin. This study identified a well developed natriuretic peptide receptor system in the urinary bladder of B. marinus but the function of the receptors remains unclear. PMID- 11884067 TI - Immunohistochemical study of androgenic gland hormone: localization in the male reproductive system and species specificity in the terrestrial isopods. AB - Androgenic gland hormone (AGH) is responsible for male sexual differentiation in crustaceans. AGH of the terrestrial isopod, Armadillidium vulgare, is a heterodimetric glycoprotein. To determine the distribution of AGH in the male reproductive system, an immunohistochemical study was carried out using antibodies raised against different components of the proAGH molecule of A. vulgare, for example, the whole molecule of recombinant proAGH expressed in Escherichia coli (E. coli-rAGH), the N-terminal nonapeptide of the B chain, and the N-terminal octapeptide of the A chain. The androgenic gland (AG) showed strong immunoreactivity to all three of these antibodies, while the testis, the seminal vesicle, and the vas deferens did not show immunostaining. To examine the species specificity of AGH, the male reproductive systems in nine species of Oniscidea were examined immunohistochemically with antibody raised against E. coli-rAGH. A positive reaction was observed in the AGs of species belonging to the Armadillidiidae, Porcellionidae, and Scyphacidae families. Immunoreactivity was strongest in A. vulgare and was stronger in Armadillidiidae than in Porcellionidae or in Scyphacidae. These results suggest that structural similarity of AGH may exist among some terrestrial isopods, although AGH seems to harbor a relatively high degree of species specificity. PMID- 11884068 TI - Molecular cloning and expression of thyroid hormone receptor alpha during salmonid development. AB - Thyroid hormones have been implicated as important regulators of teleost development. To gain a better understanding of the potential roles of the thyroid system in salmonids a genomic clone which encoded rainbow trout TR-alpha was isolated. This clone exhibited highest amino acid identity to Japanese flounder TR-alphaB (94%) and zebrafish TR-alpha1 (94%). Oligonucleotides were designed against the rainbow trout sequence and the complete coding region of Atlantic salmon TR-alpha was isolated by RACE-PCR. The Atlantic salmon sequence exhibited highest amino acid identity to rainbow trout TR-alpha (98%), Japanese flounder TR alphaB (93%), and zebrafish TR-alpha1 (90%). Atlantic salmon TR-alpha exhibited the classic modular structure associated with members of the nuclear receptor superfamily and consisted of a divergent A/B domain while the DNA and ligand binding domains were highly conserved to other teleost TR proteins. Temporal expression from the rainbow trout TR-alpha gene was monitored by semiquantitative RT-PCR at selected stages during rainbow trout embryonic and larval development. High levels of maternal transcripts were present at cleavage (Stage 6) which were rapidly degraded by gastrulation (Stage 13). Low levels of TR-alpha expression were then detected during organogenesis (Stages 20, 24, 26, 29, and 31). A peak in mRNA levels was observed at hatch (Stage 32) after which levels rose in a gradual manner during larval development (Stages 33, 34, 35, and 36) to reach maximal values at first feeding (Stage 37). These results suggest that the thyroid axis is functional and that embryonic and larval rainbow trout are at least capable of responding to thyroid hormones. These observations implicate the thyroid system as being an important regulator of salmonid development. PMID- 11884069 TI - Molecular cloning of growth hormone-encoding cDNA of an Indian major carp, Labeo rohita, and its expression in Escherichia coli and zebrafish. AB - The cDNA clones encoding for growth hormone (GH) of an Indian major carp rohu Labeo rohita were isolated from a cDNA library constructed from the poly(A)(+) RNA extracted from the pituitary glands of rohu. Partial GH cDNA of the rohu (3' end) was amplified by RT-PCR and used as probe to screen the cDNA library. Full length GH-specific cDNA clones (1180 bp) were isolated and sequenced. The sequence contains 48-bp 5'-noncoding region followed by an ORF of 621 bp and a 3' noncoding region of 521 bp. The peptide shares about 90% identity with the GH of Cyprinus carpio (Linn.) and >84% identity with GH sequences of other cyprinids. The GH-encoding cDNA of rohu has been cloned into expression vectors and GH protein has been over expressed in Escherichia coli and purified as a soluble protein. The GH cDNA was cloned into a bicistronic vector with EGFP; injection of in vitro transcribed GH-EGFP mRNA into zebrafish embryo has resulted in EGFP expression confirming the cloned GH cDNA is functional in fish and the IRES element could be effectively used in fish for bicistronic expression of foreign genes. PMID- 11884070 TI - Sight of a predator can stimulate a corticosterone response in the great tit (Parus major). AB - The corticosterone response to the sight of a natural predator was investigated in free-living and captive great tits (Parus major). Free-living great tits responded to the sight of a stuffed, slowly moving Tengmalm's owl, a major predator of great tits, with warning calls and a change in behaviour around a feeder. Great tits returned to the feeder within a few minutes and began to approach the owl, and there was no increase in plasma corticosterone levels in birds sampled 30-50 min after they first saw the owl. Captive great tits in an aviary were exposed for 30 min to a stuffed Tengmalm's owl, to a stuffed brambling, and to a cardboard box. All three stimulus objects were slowly rotated during the exposure period. Great tits exposed to the owl changed their behaviour immediately, and spent most of the time when the owl was visible flying around the aviary and hanging from the roof, with very few visits to a feeder. Great tits exposed to the brambling and to the moving box also changed their behaviour and made fewer visits to the feeder. The great tits responded to the sight of the owl with a marked increase in plasma corticosterone levels, whereas there was no change in corticosterone levels (mean levels < 11 ng/ml) in birds exposed to the brambling or to the moving box. Mean corticosterone levels were high (37.1 +/- 4.9 ng/ml) 0.5 h after exposure to the owl, remained high (38.9 +/- 6.0 ng/ml) 1 h after exposure, and had returned to basal (5.3 plus minus 1.3 ng/ml) 3 h after exposure to the owl. This is the first demonstration for any bird of a complete corticosterone response to a predator. The sight of a predator initiated a corticosterone response in great tits that could not move more than 3 m away, whereas free-living great tits that could choose how far to fly away from the predator either did not initiate a corticosterone response, or had a small corticosterone response in which corticosterone levels were not significantly different from basal 30-50 min later. The results indicate that the initiation of a corticosterone response in birds depends on whether or not a bird perceives that a stimulus is a threat. Furthermore, they illustrate the importance of not making generalised conclusions based on laboratory experiments. PMID- 11884071 TI - In vitro insulin stimulatory effect on glucose uptake and glycogen synthesis in the gills of the estuarine crab Chasmagnathus granulata. AB - The aim of the present study was to examine the effects of insulin on glucose uptake and glycogen synthesis in crab Chasmagnathus granulata gills. We observed an increased glucose uptake and incorporation of d-[(14)C]glucose into glycogen when posterior C. granulata gills were incubated in the presence of insulin; however, this was not observed in anterior gills, despite the presence of similar insulin receptors. In posterior gills, basal glucose uptake in the summer was significantly higher than in the winter. Moreover, in the summer, the insulin dose required to stimulate glucose uptake was twice as high as in the winter. However, there was no significant difference in terms of basal glycogen synthesis in summer and winter. In crustaceans, the endogenous insulin/IGFI substance might be involved in the rapid restoration of glycogen levels in the gills, increasing glucose uptake and glycogen synthesis. Bovine insulin seems to have a stimulatory effect on glycogen metabolism only in posterior gills. PMID- 11884072 TI - In vitro effect of sex steroids on cytotoxic activity of splenic macrophages in wall lizard (Hemidactylus flaviviridis). AB - Sexual dimorphism was observed in nitrite release and IL-1-like molecule production by splenic macrophages of the wall lizard (Hemidactylus flaviviridis), with a higher level in females than in males. Gonadectomy in both males and females resulted in a considerable increase of nitrite and IL-1-like molecule secretion, suggesting that the sex hormones inhibit cytotoxic activity of macrophages. To verify this assumption, dose- and time-related in vitro experiments with male and female sex steroids, dihydrotestosterone (DHT) and 17beta-estradiol (E(2)), respectively, were carried out. E(2) and DHT both significantly reduced the nitrite release and IL-1-like molecule production with an increase of dose or duration of treatment. PMID- 11884074 TI - Serotonin modulation of CHH secretion by isolated cells of the crayfish retina and optic lobe. AB - The authors used the reverse hemolytic plaque assay to investigate whether single retinal and optic lobe cells of juvenile and adult crayfish secrete crustacean hyperglycemic hormone (CHH) and whether the secretion rate depends on extracellular serotonin (5-HT) concentration. Nearly 25% of individual retinal and optic lobe cells of juvenile and adult organisms secrete CHH in response to KCl depolarization. In this condition, CHH secretion increased as a function of 5 HT concentration. In both cases, the dose-response curve indicates two different populations of CHH-secreting cells. Juveniles showed a higher CHH secretion index than did adult organisms, demonstrating a developmental interstage variation of CHH secretion. The authors conclude that (1) retinal CHH-secreting cells correspond to a population of retinal tapetal cells and (2) optic lobe CHH secreting cells correspond to two subpopulations of CHH of medulla terminalis-X organ. PMID- 11884073 TI - Free and conjugated estrogens and androgens in stallion semen. AB - The steroid content of semen from a total of 11 mature fertile stallions was studied during two breeding seasons and one winter. The levels of free and conjugated substrates (testosterone and androstenedione), and products (estradiol and estrone), of aromatase were measured by radioimmunoassay with a validated method. The results were seasonally and monthly highly variable with characteristic peaks. The concentrations of free and conjugated estrogens were always higher in the gel-free ejaculate than in the gel except in one subfertile stallion used as comparison. Furthermore, the steroid production and the maximal resulting aromatase activity, estimated by the estrogens/androgens ratio, peaked in April-May and June. The breeding season (spring and summer) presents a clear estrogenic profile with estrogens/androgens ratios higher in contrast to the nonbreeding period (autumn and winter). The involvement of estrogens in the regulation of reproduction and equine spermatogenesis is discussed, and estrogens production and thus equine aromatase is proposed as a strong marker of testicular endocrine function. PMID- 11884075 TI - The role of prolactin in fish osmoregulation: a review. AB - The protein hormone prolactin (PRL) was first discovered as an anterior pituitary factor capable of stimulating milk production in mammals. We now know that PRL has over 300 different functions in vertebrates. In fish, PRL plays an important role in freshwater osmoregulation by preventing both the loss of ions and the uptake of water. This paper will review what is currently known about the structure and evolution of fish PRL and its mechanisms of action in relation to the maintenance of hydromineral balance. Historically, functional studies of fish PRL were carried out using heterologous PRLs and the results varied greatly between experiments and species. In some cases this variability was due to the ability of these PRLs to bind to both growth hormone and PRL receptors. In fact, a recurring theme in the literature is that the actions of PRL cannot be generalized to all fish due to marked differences between species. Many of the effects of PRL on hydromineral balance are specific to euryhaline fish, which is appropriate given that they frequently experience sudden changes in environmental salinity. Much of the recent work has focused on the isolation and characterization of fish PRLs and their receptors. These studies have provided the necessary tools to obtain a better understanding of the evolution of PRL and its role in osmoregulation. PMID- 11884076 TI - Prominent expression of transforming growth factor beta2 gene in the chicken embryonic gonad as revealed by suppressive subtraction cloning. AB - cDNA cloning from chicken embryonic gonad subtracted from tissues of the brain, heart, liver, gizzard, mesonephros, and muscle was performed to identify growth factor genes with expression unique to embryonic ovary and testis. We obtained several cDNA clones encoding known and many unknown genes. We found for the first time that the transforming growth factor beta2 (TGF-beta2) is preferentially expressed in the chicken embryonic ovary and testis. cDNA subtraction cloning with respect to the selective expression of TGF-beta2 in the ovary and testis was further analyzed by reverse transcription-polymerase chain reaction analyses of other embryonic tissues. The ontogeny of TGF-beta2 was evaluated in chicken embryonic ovary and testis. In both testis and ovary, the levels of TGF-beta2 transcripts were high during the early period of embryonic development (E7), gradually decreased until the late embryonic days (E14--E17), and then slightly increased at the last embryonic day (E21). There was no difference in the TGF beta2 transcripts per RNA between the left and the right ovaries. TGF-beta2 may have a critical role in the regulation of the development of chicken ovarian and testicular germ cells during the embryonic period. PMID- 11884077 TI - Central administration of corticotropin-releasing hormone stimulates locomotor activity in juvenile chinook salmon (Oncorhynchus tshawytscha). AB - This study evaluated the effect of corticotropin-releasing hormone (CRH) on locomotor activity, habitat choice, and social behavior in juvenile spring chinook salmon (Oncorhynchus tshawytscha). An intracerebroventricular (ICV) injection of CRH caused a dose-dependent increase in locomotor activity. The stimulatory effect of exogenous CRH on locomotor activity lasted for at least 24 h. Injection (ICV) of a peptide antagonist of CRH, alpha-helical CRH(9-41) (ahCRH), prevented the increase in locomotor activity when administered concurrently with CRH. Furthermore, fish administered the antagonist alone had significantly lower locomotor activity levels compared to saline-injected control fish. The effects of CRH are often dependent on the social context. However, no evidence was found that the presence of conspecifics during the testing procedure affected locomotor activity following ICV injections of CRH. Similarly, ICV injections of CRH or ahCRH did not have a significant effect on the mean time spent in contact with a conspecific. However, the position of fish in the tank was affected by the treatments. ICV injections of CRH significantly increased the amount of time that fish spent near the center of the tank. Furthermore, ICV injections of ahCRH significantly increased the mean time taken for fish to find cover in the tank. The effect of CRH and ahCRH on locomotor activity was not related to changes in plasma cortisol or thyroxine. These results support the hypothesis that endogenous CRH within the central nervous system is involved in the stimulation of locomotor activity in fish. Furthermore, CRH may also alter habitat choice in a novel environment. PMID- 11884078 TI - Isolation and characterization of a homologue of mammalian prolactin-releasing peptide from the tilapia brain and its effect on prolactin release from the tilapia pituitary. AB - In the tilapia (Oreochromis mossambicus), as in many teleosts, prolactin (PRL) plays a major role in osmoregulation in freshwater. Recently, PRL-releasing peptides (PrRPs) have been characterized in mammals. Independently, a novel C terminal RF (arginine-phenylalanine) amide peptide (Carrasius RF amide; C-RFa), which is structurally related to mammalian PrRPs, has been isolated from the brain of the Japanese crucian carp. The putative PrRP was purified from an acid extract of tilapia brain by affinity chromatography with antibody against synthetic C-RFa and HPLC on a reverse-phase ODS-120 column. The tilapia PrRP cDNA was subsequently cloned by polymerase chain reaction. The cDNA consists of 619 bp encoding a preprohormone of 117 amino acids. Sequence comparison of the isolated peptide and the preprohormone revealed that tilapia PrRP contains 20 amino acids and is identical to C-RFa. Incubation of the tilapia pituitary with synthetic C RFa (100 nM) significantly stimulated the release of two forms of tilapia PRL (PRL188 and PRL177). However, the effect of C-RFa was less pronounced than the marked increase in PRL release in response to hyposmotic medium. The ability of C RFa to stimulate PRL release appears to be specific, since C-RFa failed to stimulate growth hormone release from the pituitary in organ culture. In contrast, rat and human PrRPs had no effect on PRL release. C-RFa was equipotent with chicken GnRH in stimulating PRL release in the pituitary preincubated with estradiol 17beta. Circulating levels of PRL were significantly increased 1 h after intraperitoneal injection of 0.1 microg/g of C-RFa in female tilapia in freshwater but not in males. These results suggest that C-RFa is physiologically involved in the control of PRL secretion in tilapia. PMID- 11884079 TI - Identification and properties of a Gs protein in catfish liver membranes. AB - The presence of G proteins and their involvement in adrenergic signaling has been investigated in catfish (Ictalurus melas) liver membranes. Adenylyl cyclase activity was potently stimulated by the nonhydrolyzable analog of GTP, [35S]guanosine 5'-O-(gamma-thiotriphosphate) (GTPgammaS) (maximal activation of about eightfold at 10(-5) M; half-maximal activation at 1.31 x 10(-7) M), and reduced by the competitive inhibitor of GTP, GDPbetaS (70% maximal inhibition at 10(-4) M; half-maximal inhibition at 1.98 x 10(-7) M). Forskolin dramatically enhanced enzyme activity (up to about 3500% at 100 microM), and its action was not affected by guanine nucleotides, confirming that the diterpene effect occurred only at targets downstream of the G proteins. Receptor-dependent G protein activity was evaluated by a [(35)S]GTPgammaS binding assay. At 100 microM GDP, 100 mM NaCl, and 5 mM MgCl2, after an incubation of 90 min at 20 degrees, a Kd of 18.6 nM and a Bmax of 105.7 pmol/mg protein for [35S]GTPgammaS binding to catfish liver membranes were determined. The binding of the tracer was enhanced by 1 microM epinephrine, up to a maximum of 158%, and inhibited by NF 449, a G(s)alpha-selective antagonist with half-maximal effect in the micromolar range. Immunoblotting analysis with a specific anti-G(s)alpha antibody revealed a single band of about 45 kDa mass. This result represents the first demonstration of the presence of G protein alpha(s) subunits in the liver of an ectothermal vertebrate. PMID- 11884080 TI - Zebrafish as a model for vertebrate reproduction: characterization of the first functional zebrafish (Danio rerio) gonadotropin receptor. AB - Vertebrate reproduction is tightly regulated by conserved glycoprotein hormones produced by the pituitary gland. Follicle-stimulating hormone (FSH) in tetrapods and gonadotropic hormone I (GTH-I) in fishes are orthologous glycoprotein hormones that control the timing of egg production and the number of eggs produced. Zebrafish, a well-established genetic model for developmental biology, also offers potential advantages for studies of reproductive toxicology, especially for modeling the impact of pollutants on fish reproductive processes. To facilitate these studies we have identified, expressed, and characterized the zebrafish GTH-I receptor. This receptor (zfGTHR-I)exhibits strong sequence similarity to the tetrapod FSH receptors and to GTHR-I from salmon and catfish. Human 293 cells transfected with zfGTHR-I exhibit increased cAMP levels after treatment with carp pituitary extracts or human FSH, but not when treated with a ligand to a related receptor (human chorionic gonadotropin). Northern blotting and RT-PCR analyses indicate that zfGTHR is expressed in ovaries from sexually mature fish, but not in immature fish. Several alternative splice variants of the receptor affecting putative exons 2-4 that encode dramatically shortened receptor fragments lacking the transmembrane domain as well as regions previously implicated in ligand binding were identified by RT-PCR. The zfGTHR-I sequence opens the way to study effects of genetic mutations or chemicals on ovarian zfGTHR-I expression and function in zebrafish. PMID- 11884081 TI - Development of a time-resolved fluoroimmunoassay for insulins and its application to monitoring of insulin secretion induced by feeding in the barfin flounder, Verasper moseri. AB - A time-resolved fluoroimmunoassay (TR-FIA) system was developed to quantify insulin levels in the barfin flounder. This TR-FIA system is a solid-phase assay based on competition of unlabeled insulins and biotinylated barfin flounder insulin-II against an anti-barfin flounder insulin-II antibody. The minimum detectable level of barfin flounder insulin-I and -II in this TR-FIA was 10 pg/well which corresponded to 1.0 ng/ml, and insulin-II showed slightly higher crossreactivity than insulin-I. The accuracy of this TR-FIA was assured by specificity test, validation test, and recovery test using plasma added insulin II. The results indicated the high specificity and sufficient accuracy of this assay system for insulin level measurement. This system was applied to the measurement of plasma insulin levels of fed and fasted barfin flounders. Plasma insulin levels (average +/- SEM) in fed flounders reached a maximum 2 h (9.3 +/- 1.7 ng/ml) and decreased gradually thereafter, while those in fasted flounders remained at low levels (1.1 +/- 0.1-2.0 +/- 0.2 ng/ml) during the experiment. After removing proteins by acidification and subsequent gel filtration, plasma samples taken from fed and fasted flounders at 2 h after feeding were fractionated separately by reversed-phase HPLC. In fed flounders, insulin immunoreactivity was detected in fractions corresponding to those of insulin-I or -II. The ratio of integrated insulin immunoreactivities of each peak was 0.378 +/ 0.044 (average +/- SD). This value was in good agreement with those (0.355 +/- 0.019) of absorbance areas of each insulin from Brockmann body extracts of the barfin flounder on reversed-phase HPLC. In fasted flounders, very weak insulin immunoreactivities were observed at retention times corresponding to those of insulin-I and -II. These results indicated that both insulin-I and -II were secreted into the blood being induced by feeding stimulation with approximately the same ratio as that of the quantities harbored in the Brockmann body. PMID- 11884082 TI - Cloning and gene expression of a cDNA for the chicken follicle-stimulating hormone (FSH)-beta-subunit. AB - Follicle-stimulating hormone (FSH) is a member of pituitary glycoprotein hormones that are composed of two dissimilar subunits, alpha and beta. Very little information is available regarding the nucleotide and amino acid sequence of FSH beta in avian species. For better understanding of the phylogenic diversity and evolution of FSH molecule, we have isolated and sequenced the complete complementary DNA (cDNA) encoding chicken FSH-beta precursor molecule by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) methods. The cloned chicken FSH-beta cDNA consists of 2457-bp nucleotides, including 44-bp nucleotides of the 5'-untranslated region (UTR), 396 bp of the open reading frame, and an extraordinarily long 3'-UTR of 2001-bp nucleotides followed by a poly(A)((16)) tail. It encodes a 131-amino-acid precursor molecule of FSH-beta-subunit with a signal peptide of 20 amino acids followed by a mature protein of 111 amino acids. Twelve cysteine residues, forming six disulfide bonds within beta-subunit and two putative asparagine linked glycosylation sites, are also conserved in the chicken FSH-beta-subunit. Four proline residues, presumably responsible for changing the backbone direction of protein structure, are conserved in chicken FSH-beta-subunit as well. The nucleotide sequence of chicken FSH-beta cDNA shows high homology with quail FSH beta cDNA, 97% homology in the open reading frame, and 85% homology in the 3' UTR. The deduced amino acid sequence of chicken FSH-beta-subunit shows a remarkable similarity to other avian FSH-beta-subunits, 98% homology with quail, and 93% homology with ostrich, whereas a lower similarity (66 to 70%) is noted when compared with mammalian FSH-beta-subunits. By contrast, when comparing with the beta-subunits of chicken luteinizing hormone and thyroid-stimulating hormone, the homologies are as low as 37 and 40%, respectively. FSH-beta mRNA was only expressed in pituitary gland out of various tissues examined and can be up regulated by gonadotropin-releasing hormone in pituitary tissue culture as estimated by real-time quantitative PCR. PMID- 11884083 TI - Characterization of outer ring iodothyronine deiodinases in tissues of the saltwater crocodile (Crocodylus porosus). AB - The distribution and characterization of outer ring deiodination (ORD) using reverse triiodothyronine (rT3) and thyroxine (T4) as substrates is reported in microsomes of liver, kidney, lung, heart, gut, and brain tissues from juvenile saltwater crocodiles (Crocodylus porosus). In lung and heart only small amounts of rT3 ORD and T4 ORD were detected, while in brain only a small amount of T4 ORD was detected. More detailed characterization studies could be performed on liver, kidney, and gut microsomes. Reverse T3 outer ring deiodination (rT3 ORD) was the predominant activity in liver and kidney microsomes. The properties of crocodile liver and kidney rT3 ORD, such as preference for rT3 as substrate, a dithiothreitol (DTT) requirement of 10 mM, inhibition by propylthiouracil (PTU), and Michaelis-Menten (Km) constant in the micromolar range, correspond to the properties previously reported for a type I deiodinase. The temperature optimum for rT3 ORD was between 30 and 35 degrees. There was also rT3 ORD activity in gut microsomes, along with what appeared to be a type II-like, low-Km deiodinase with a substrate preference for T4. There was also a small amount of T4 ORD activity in liver and kidney microsomes. Liver T4 ORD, like a type II deiodinase, had a preference for T4 as substrate at low substrate concentrations and a DTT requirement of 15 mM and was insensitive to PTU. However, at high substrate concentrations the predominant activity was of the type I deiodinase nature. T4 ORD in liver had an optimal incubation temperature of 30 to 35 degrees. Gut microsomal T4 ORD was also type II-like at low substrate concentrations and type I-like at high substrate concentrations. Gut T4 ORD had an optimal incubation temperature of 25 to 30 degrees and a DTT requirement of 20 mM DTT. Kidney microsomal T4 ORD had the same optimal temperature and DTT requirement as that in gut microsomes; however, there was no competition by low substrate concentrations. These results suggest that ORD in juvenile saltwater crocodile kidney is most likely exclusively catalyzed by a type I-like deiodinase. Liver and gut ORD, in contrast, is catalyzed by two enzymes, with a predominance of a type I-like deiodinase in liver and a type II-like deiodinase in gut. Low-Km T3 IRD activity could not be detected in any tissues of the juvenile saltwater crocodile. PMID- 11884084 TI - Factors affecting plasma concentrations of prolactin in the common eider Somateria mollissima. AB - In the common eider only the females incubate while they fast for 25 days. Thus, since they rely entirely on their body reserves for successful incubation, they can be defined as capital incubators. To assess the potential effects of their initial body mass, the incubation duration, and depletion in body reserves on prolactinemia, blood samples of eiders were analyzed during the breeding cycle and an experimental manipulation of the duration of incubation. Levels of circulating prolactin increased at the onset of incubation and then reached a high and stable level during incubation before increasing sharply before hatching. The prolactin level decreased significantly upon hatching. Captive females deprived from their eggs exhibited a rapid decrease in prolactinemia, suggesting that egg stimuli are necessary to prolactin secretion. Aunts, i.e., helper females caring for conspecific young, presented prolactin levels higher than nonbreeding captive females but not significantly different from those of females at hatching. Plasma prolactin at hatch was directly related to body mass loss. Birds with shortened incubation have higher body masses and showed higher levels of prolactinemia at hatching than the control group, in accordance with the idea that circulant prolactin at hatching is linked to body condition. Females which underwent an extended incubation (and started to eat again) displayed a low body mass and a high prolactinemia. These data therefore suggest that refeeding, albeit increasing the risk of predation, enhances prolactin secretion and allows the bird to continue incubation despite that it has reached a poor body condition. PMID- 11884085 TI - Immunocytochemical demonstration of melanotropic and adrenocorticotropic cells from the gilthead sea bream (Sparus aurata L., Teleostei) by light and electron microscopy: an ontogenic study. AB - In the pituitary of gilthead sea bream, Sparus aurata, melanotropic (MSH) and adrenocorticotropic (ACTH) cells were identified at the light and electron microscopic levels using rabbit anti-synthetic alphaMSH (MSH) and anti-human ACTH (1-24) (ACTH) sera. The distribution of these cell types was followed from hatching to 48 months. The techniques used included the peroxidase anti peroxidase (PAP) method, conventional electron microscopy, and an immunogold technique. Using PAP, MSH (immunoreactive to both anti-MSH and anti-ACTH) and ACTH (immunoreactive to anti-ACTH) cells were detected from hatching onward. These cells were distinguished ultrastructurally in 1-day-old larvae. Immunogold labeling was first detected in MSH cells in 5-day-old larvae, while ACTH cells were only immunogold labeled in adults. In newly hatched larvae, MSH cells were located from the middle to the posterior region of the adenohypophysis, while ACTH cells were found in the dorsoanterior region, next to the hypothalamus. At this age, both cell types were scarce. As the fish developed, these cell types progressively increased in number: MSH cells made up a layer surrounding the neurohypophysis (NH) in the pars intermedia (pi), whereas ACTH cells bordered the developing NH in the rostral pars distalis (rpd). From 82 days onward, a few MSH cells were observed in the proximal pars distalis (ppd) next to the pi and some ACTH cells were seen in the ppd next to the rpd. In adult specimens, both MSH and ACTH cells were adjacent to the stellate cells and showed processes and synaptic like structures. MSH cells exhibited numerous round secretory granules with a granular content and of varying electron density and compactness. These granules were immunogold labeled with anti-MSH serum. Electron-dense secretory granules near the Golgi complex immunoreacted with anti-MSH, anti-ACTH, or with both antisera. ACTH cells exhibited round secretory granules with a homogeneous, high electron-dense core and a narrow, clear halo. These granules immunoreacted with anti-ACTH serum. The main ultrastructural features that characterize the MSH and ACTH cells of adults appeared early during ontogeny. Involutive MSH and ACTH cells were only observed in adult specimens. PMID- 11884086 TI - Seasonal changes in testicular steroidogenesis in the toad Bufo arenarum H. AB - The biosynthesis of androgens in Bufo arenarum takes place through the 5-ene pathway that includes 5-androstane-3beta,17beta-diol as intermediate in testosterone biosynthesis. Besides testosterone and 5alpha-dihydrotestosterone, testes are able to synthesize 5alpha-pregnan-3,20-dione and several 3alpha- and 20alpha-reduced derivatives. Steroid biosynthesis changes during the breeding period (spring and early summer), turning from androgen to C21 steroid production. During the reproductive season, the production of progesterone, 5alpha-pregnan-3alpha,20alpha-diol, 3alpha-hydroxy-5alpha-pregnan-20-one, and 5alpha-pregnan-3,20-dione increases significantly. The function of most of these steroids in amphibians remains unknown. However, 5alpha-androstan-3alpha,17beta diol and 3alpha-hydroxy-5alpha-pregnan-20-one were shown to be neuroactive in mammals, modulating sexual behavior. Thus, 5alpha/3alpha-reduced steroids could be involved in the regulation of the reproductive behavior in B. arenarum, a species with a dissociated reproductive pattern. Percentage contribution of each enzymes to the total metabolism reveals that neither 3beta-hydroxysteroid dehydrogenase/isomerase nor 5alpha-reductase change throughout the reproductive cycle. However, a strong reduction in 17-hydroxylase-C(17-20) lyase activity occurs in the reproductive season, suggesting that this enzyme could represent a key enzyme in the regulation of the seasonal change of steroidogenesis. Also, 3alpha-hydroxysteroid dehydrogenase and 20-hydroxysteroid dehydrogenase activities increase during the reproductive period, implying that steroid metabolism is clearly focused on C21-reduced steroids. PMID- 11884088 TI - Development of animal recognition: a difference between Parts and Wholes. AB - A series of experiments examined children's recognition of animals by their features (Parts) and by the relative scale of the parts (Wholes). They were asked to identify the correct picture of an animal they could name from the original plus two computer-generated alternatives. We examined the developmental trends associated with upright (Studies 1 and 3) and inverted presentations (Study 3). Both experiments confirmed children's superior ability in dealing with the recognition of animal Parts over animal Wholes, especially for the younger ages tested (6- and 10-year-olds). It was not until the ages of 15-16 that children demonstrated equal performance on Whole and Part items. The late acquisition of animal Whole recognition is compared to the late acquired configural skills proposed for face recognition. PMID- 11884087 TI - Melanocyte-stimulating hormone plasma levels and environmental illumination in the cuttlefish, Sepia officinalis: a role for the neurosecretory system of the vena cava in cephalopods. AB - A melanotropin-like peptide (alpha-melanocyte-stimulating hormone or alpha-MSH) is suggested to be released into the circulatory system of cephalopods via the neurosecretory system of the vena cava or NSV, where neurosecretory vesicles contained within the axons of the NSV-neuropil on the inner surface of the vena cava lie in close contact with the venous circulation. Radioimmunoassay of blood plasma samples taken from the cephalic vein of anaesthetised cuttlefish, Sepia officinalis showed that immunoreactive alpha-MSH (ir alpha-MSH) was detectable within the cuttlefish circulatory system. The validity of the assay for determination of cuttlefish ir alpha-MSH was determined by parallelism of the alpha-MSH standard curve against serially diluted cuttlefish plasma samples. Plasma samples taken during a natural day-night-day illumination cycle showed a significant elevation in ir alpha-MSH concentration to 1.44 +/- 0.26 ng ml(-1) during the middle of the dark phase compared to concentrations of 0.48 +/- 0.13 and 0.35 +/- 0.10 ng ml(-1) in the middle of the light phases of the illumination cycle. So far, indirect evidence suggests Sepia officinalis may modulate chromatophore activity, body patterning, and behaviour via neuroendocrine release and circulating titres of this proopiomelanocortin-derived peptide. PMID- 11884089 TI - Developmental and content effects in reasoning with causal conditionals. AB - Two predictions derived from Markovits and Barrouillet's (2001) developmental model of conditional reasoning were tested in a study in which 72 twelve-year olds, 80 fifteen-year-olds, and 104 adults received a paper-and-pencil test of conditional reasoning with causal premises ("if cause P then effect Q"). First, we predicted that conditional premises would induce more correct uncertainty responses to the Affirmation of the consequent and Denial of the antecedent forms when the antecedent term is weakly associated to the consequent than when the two are strongly associated and that this effect would decrease with age. Second, uncertainty responding to the Denial of the antecedent form ("P is not true") should be easier when the formulation of the minor premise invites retrieval of alternate antecedents ("if something other than P is true"). The results were consistent with the hypotheses and indicate the importance of retrieval processes in understanding developmental patterns in conditional reasoning with familiar premises. PMID- 11884090 TI - The role of cues to differential absolute size in children's transitive inferences. AB - To investigate the role that "nonlogical" cues might play in transitive inference, 6- and 7-year-olds were given a three-term transitive task in which perceptual cues to differential absolute size were either present or absent. Relationships between the taught premises and the relational information that was physically present were manipulated using four basic conditions: "congruent," "inverse," "pretended," or "persuaded." Both age groups showed identical overall premise memory, but the younger group tended to reason more on the basis of the perceptual information rather than on the successfully encoded premise information. Contrasts between the various conditions showed that categorical effects can be circumvented in three-term problems with appropriate controls, that there may be qualitative as well as quantitative differences in transitive inference with age, and that transitive inference is not based solely on memory. The findings also indicate that, although 7-year-olds are competent in "logic based" transitive inference, they experience great difficulty on tasks involving pretend information. PMID- 11884091 TI - The development of explicit memory for basic perceptual features. AB - In three experiments with 164 individuals between 4 and 80 years old, we examined age-related changes in explicit memory for three perceptual features--item identity, color, and location. In Experiments 1-2, feature recognition was assessed in an incidental learning, gamelike task resembling the game Concentration. In Experiment 3, feature recognition was assessed using a pencil and-paper task after intentional learning instructions. The form of the explicit memory function across the life span varied with the particular perceptual feature tested and the type of task. Item recognition was excellent at all ages but was significantly poorer for older adults than children, color recognition peaked in late childhood on the gamelike task, and location recognition peaked in early adulthood on the pencil-and-paper task. These findings indicate that performance on explicit memory tests is not a consistent inverted U-shaped function of age across various features. Explicit memory performance depends on what is measured and how. Because explicit memory typically reflects a composite of different features, age-related changes in explicit memory will not necessarily correspond to the function for any single one. PMID- 11884092 TI - The development of organizational strategies in children: evidence from a microgenetic longitudinal study. AB - The authors examined memory on a sort-recall task in children 8 to 12 years of age. Children were first classified as either strategic or nonstrategic on a sort recall pretest and then participated in an 11-week microgenetic study involving nine sessions. Strategy use was assessed on each trial. Consistent with past longitudinal research, changes from nonstrategic to strategic behavior occurred suddenly rather than gradually. Once children began using organizational strategies, their recall performance improved immediately. Deliberate strategy use was clearly reflected by sorting behavior during encoding but not in clustering during recall. Conclusions about whether there are children with utilization deficiencies are thus affected by how the concept is defined and whether sorting or clustering is taken as the indicator of spontaneous strategy use. PMID- 11884093 TI - Effects of the "beauty is good" stereotype on children's information processing. AB - The authors tested schematic information processing as a function of attractiveness stereotyping in two studies. An adult experimenter read children (ages 3 to 7 years) eight different stories in which a child narrator encountered two characters who varied in level of attractiveness and displayed positive or negative traits that were either consistent or inconsistent with the "beauty is good" stereotype. Following the story, the experimenter showed each child a photograph of the two characters' faces and asked the child to point to the character who displayed the positive trait. In Experiment 1, children made more errors in identifying female characters with stereotype inconsistent traits but did just the opposite with male characters. Experiment 2 replicated the findings with female characters but found no difference in errors with male characters. The findings have implications for how attractiveness and gender stereotypes affect children's information processing, how attractiveness schemata may be organized, and why physical attractiveness stereotypes are maintained. PMID- 11884094 TI - Generalized imitation within three response classes in typically developing infants. AB - Effects of modeling and contingent praise on infant imitation of three different responses was analyzed. Generalization to nonreinforced probe models was assessed both within and across response types. Three 12- to 14-month-old infants and their mothers participated in this study. During baseline the mothers provided models only. During treatment mothers modeled and also praised contingent upon infant matching of the training models. During interspersed probe trials the mothers modeled different responses, which, if matched by the infant, produced no praise. The three responses modeled were motor-with-toy, motor-without-toy, and vocal responses. The dependent measure was the percentage of maternal models that were matched by the infant within 6 s. Nonmatching responses of the same response type were also measured. Results showed a systematic increase in the percentages of training and probe models matched by the three infants following the introduction of the model-and-praise treatment condition. Nonmatching responses did not systematically increase. Thus, imitation generalized within response class, but not across response classes. PMID- 11884095 TI - Young infants' perception of unity and form in occlusion displays. AB - Young infants have been reported to perceive the unity of a center-occluded object when the visible ends of the object are aligned and undergo common motion but not when the edges of the object are misaligned (Johnson & Aslin, 1996). Using a recognition-based paradigm, the authors investigated the possibility that past research failed to provide sufficiently sensitive assessments of infants' perception of the unity of misaligned edges in partial occlusion displays. Positive evidence was obtained in 4-month-olds for veridical perception of the motion and location of a hidden region but not its orientation, whereas 7-month olds, in contrast to the younger infants, appeared to respond to the orientation of the hidden region. Overall, the results suggest that habituation designs tapping recognition processes may be particularly efficacious in revealing infants' perceptual organization. In addition, the findings provide corroborative evidence for the importance of both motion and orientation in young infants' object segregation and for the difficulty in achieving percepts of the global form of a partly occluded object. PMID- 11884097 TI - Hand transplantation. PMID- 11884098 TI - Hand transplantation: ethics, immunosuppression and indications. PMID- 11884099 TI - Hand transplantation--risks of immunosuppression. PMID- 11884100 TI - My reflections and opinions on hand transplantation. PMID- 11884101 TI - Hand transplantation--an opinion. PMID- 11884103 TI - Some aspects of hand transplantation. PMID- 11884102 TI - Bilateral hand transplantation--indication and rationale. PMID- 11884104 TI - Aesthetic hand prosthesis: gadget or therapy? Presentation of a new classification. AB - Aesthetic prostheses must fulfill a functional as well as an aesthetic role. The function of these prostheses may be perceived as an additional passive support, or as an instrument of social function and an aid to the patient. This functional role has been observed in the continued use of these prostheses over several years. We reviewed our experience with prescription, fitting and follow-up care in 2847 patients. Classification of these patients was based on their age, level of amputation (with or without functional pinch) and cause of amputation. Analysis of the data ratifies our indications for prosthesis fitting. The long term wearing of these aesthetic prostheses confirms their use as therapeutic tools. PMID- 11884105 TI - Open carpal tunnel decompression in long-term haemodialysis patients [corrected]. AB - This retrospective study assessed the treatment of 91 cases of carpal tunnel syndrome in long-term haemodialysis patients. One group of patients underwent an enlargement reconstruction of the flexor retinaculum with synovectomy and the other group was treated with a conventional carpal tunnel release. There were no major changes or differences between the outcomes of the two groups. However, there was an earlier functional recovery of grip strength and a lower recurrence rate in the enlargement plasty with synovectomy group. PMID- 11884106 TI - Carpal tunnel syndrome in radial dysplasia. AB - We treated three patients for carpal tunnel syndrome which developed more than 10 years after reconstructive surgery for radial dysplasia. All responded to decompressive surgery. The radial carpal bones were hypoplastic in all cases, and in two we measured the carpal tunnel with computed tomography (CT). This showed that the anteroposterior diameter and cross-sectional area of the carpal tunnel were small because of the hypoplasia of the carpal bones. We believe carpal tunnel syndrome occurs with radial dysplasia because of the narrow anteroposterior diameter and small cross-sectional area of the carpal tunnel. PMID- 11884107 TI - The treatment of dorsal fracture-dislocation of the proximal interphalangeal joint by closed reduction and Kirschner wire fixation: a 16-year follow up. AB - Ten patients who had sustained 11 unstable dorsal fracture-dislocations of finger proximal interphalangeal joints were reviewed at a mean follow-up of 16 years. All had been treated acutely by closed reduction and transarticular Kirschner wire fixation of the proximal interphalangeal joint, without any attempt at reduction of the fracture of the base of the middle phalanx, which probably involved 30-60% of the articular surface. Seven of the ten patients complained of no finger pain or stiffness, and none complained of severe pain. There was a mean fixed flexion deformity of 8 degrees at the proximal interphalangeal joint, which had a mean arc of movement of 85 degrees. Although subchondral sclerosis and mild joint space narrowing were observed in some instances, there were no severe degenerative changes. These results confirm that this technique is a reliable treatment method for these injuries, and produces satisfactory long-term results. PMID- 11884108 TI - The innervation of the proximal interphalangeal joint and its application in neurectomy. AB - In an anatomic study of 64 fingers, we demonstrated that the proximal interphalangeal joint is innervated by branches of the palmar digital nerves. The number of articular branches ranges from two to four and their origins from the digital nerve are between 2 and 8 mm from the proximal interphalangeal joint. In a clinical series of 24 neurectomies in 21 patients, there was a significant improvement in pain and range of motion in 22 fingers. We conclude that neurectomy is a therapeutic option in the treatment of osteoarthritis of the proximal, interphalangeal joint. PMID- 11884109 TI - A comparison of the findings of wrist arthroscopy and magnetic resonance imaging in the investigation of wrist pain. AB - Between 1996 and 1999, 54 patients with wrist pain had magnetic resonance imaging performed using a 1.5 Tesla scanner without a wrist coil. Wrist arthroscopy was performed using a standard technique. The findings were then compared. Magnetic resonance imaging had a low sensitivity for the detection of triangular fibrocartilage complex injuries (0.44) and scapholunate ligament injuries (0.11) when wrist arthroscopy was used as the standard of reference. We conclude that when a magnetoresonance technique that does not employ a dedicated wrist coil is used, a negative magnetic resonance imaging scan does not exclude these two significant injuries. PMID- 11884110 TI - Arthroscopic diagnosis and treatment of dorsal wrist ganglion. AB - Thirty-seven patients with dorsal wrist ganglia underwent arthroscopic resection. The mean follow-up was 20 months, and no complications were encountered. The ganglia were classified into three types according to their arthroscopic appearance. This classification helps to determine the amount of dorsal capsular resection required. PMID- 11884111 TI - Mechanical strength of intramedullary pinning and transfragmental Kirschner wire fixation for Colles' fractures. AB - Four methods of distal radius fracture fixation were tested in this experimental biomechanical study of an unstable Colles' fracture model. Sixty artificial radial bones and seven pairs of cadaveric radii were used. Seven additional pairs of paired cadaveric radial epiphyses were used in separate perforation and cut out tests. Tests with the artificial bones showed that the rigidity of the four tested fixation methods was comparable, except for proximal intramedullary pinning. The rigidity of the NODE fixation in the cadaveric radii was greater than that of transfragmental pinning. The load at failure was significantly greater for the NODE system than for the Kirschner wire models. Rigidity and failure loads showed a positive correlation with bone mineral density. The current study indicates the NODE system was the strongest of the fixation methods. PMID- 11884112 TI - Surgical treatment for recurrent dislocation of the extensor carpi ulnaris tendon. AB - Twelve patients with recurrent dislocation of the extensor carpi ulnaris tendon were treated with repair or reconstruction of its tendon sheath, and each had a satisfactory result. We found three types of disruption of the fibro-osseous sheath. Type A: the fibro-osseous sheath ruptured ulnarly and the torn sheath lay superficial to the tendon (n=5). These were treated by reconstruction of the sheath using a piece of the extensor retinaculum. Type B: the fibro-osseous sheath ruptured radially and the torn sheath lay in the ulnar groove beneath the tendon (n=3). These were treated by direct suture of the sheath over the tendon. Type C: detachment of the periosteum from the ulnar side of the ulna in continuity with the fibro-osseous sheath formed a false pouch into which the tendon easily dislocated (n=4). These were treated by reattachment of the periosteum. PMID- 11884113 TI - Surgical correction of extensor tendon subluxation and ulnar drift in the rheumatoid hand: long-term results. AB - Subluxation of the extensor digitorum communis tendons in the rheumatoid hand causes ulnar digital drift. If passively correctable, the digit may be realigned by soft tissue rebalancing and extensor centralization, which may preserve a more functional arc of motion than achieved with arthroplasty. A total of 71 centralization procedures were done in 15 rheumatoid patients with a mean age of 55 years and an average follow-up of 9 years. A distally based central-third strip of extensor tendon was used. Correction of ulnar drift deformity was from an average of 47 degrees preoperatively to 7.9 degrees postoperatively, and correction of active range of motion of the metacarpophalangeal joints was from an average of 38 degrees to 56.2 degrees. Reoperation and complication rates were low. This technique corrects and maintains ulnar drift in the rheumatoid hand. Range of motion at the metacarpophalangeal joint level is improved and converted to a more functional one by decreasing the extensor lag. PMID- 11884114 TI - The value of crossed intrinsic transfer after metacarpophalangeal silastic arthroplasty: a comparative study. AB - Seventy three hands in patients with rheumatoid arthritis undergoing primary index to small finger metacarpophalangeal joint replacements were studied retrospectively. In twenty eight hands a crossed intrinsic transfer was performed and in forty five hands it was not. A similar splintage and rehabilitation programme was followed in each group. The two treatment groups had similar preoperative ulnar drift (crossed intrinsic transfer group mean 27 degrees, comparative group 29 degrees). At a mean follow up of 50 months the crossed intrinsic transfer group had statistically less ulnar drift (crossed intrinsic transfer group mean 6 degrees, comparative group mean 14 degrees, P=0.01). There were no other significant differences at follow up. PMID- 11884115 TI - Long-term results of digital arthrodesis with the Harrison-Nicolle peg. AB - This study determined the long-term success of digital arthrodesis with the Harrison-Nicolle peg. We reviewed 90 digital joints in 60 patients fused with the peg between 1986 and 1998 at a mean follow-up of 6 (range 2-11) years. The prime indication for surgery was rheumatoid arthritis. The early complication rate was 8%. At 1 month 89% of joints were pain-free and stable. In the long-term follow up, 96% of the joints were pain-free and stable, with the original angle of fusion. 85% achieved bony fusion, with no clinical difference between bony and fibrous fusion. Overall there was a significantly higher complication rate in the distal interphalangeal joint. We conclude that, with the exception of the distal interphalangeal joint, the Harrison-Nicolle peg is extremely effective for digital arthrodesis in the rheumatoid patient. PMID- 11884117 TI - A new technique to salvage digital replantation with compromised venous outflow. AB - The use of an intravenous cannula to salvage digital replantation with compromised venous outflow is described. The advantages of this technique when compared to other methods used to treat "artery only" replantations are discussed. PMID- 11884116 TI - Indications and selection for digital amputation and replantation. AB - Even though replantation surgery has now become a routine procedure, it remains delicate and demanding surgery, requiring adequate training and expertise in microsurgical techniques. Well-defined selection criteria for replantation procedures have evolved over the past few years, including definitive guidelines for thumb, single digit, multiple digit and mid-palm amputations. For more complex cases, other techniques, including transpositional microsurgery and various secondary reconstructive procedures, such as toe-to-hand transfer, are now available. Although replantation procedures have been simplified, a second surgical team can save valuable surgical time by debriding and identifying the vessels in the amputated part, harvesting microvenous grafts, and performing bone fixation or tendon repair among other things, while the chief surgeon focuses on revascularization. Overall, the most significant guideline underlining the philosophy of digital replantation today reflects the aim of not only ensuring the survival of a digit, but its functional use as well. Experience dictates that this can be achieved only if the basic principles and indications of replantation surgery are adhered to. PMID- 11884118 TI - Finger sucking digital deformities. AB - We report our experience with severe digital deformities caused by prolonged finger sucking. Our analysis of nine patients demonstrated that the deformity was mainly located in the proximal and middle phalanges of the affected digit(s). We have reviewed the literature, enumerated the common patterns of "finger sucking" and explained the resulting deformities. Corrective osteotomy, preferably at the metacarpal base level, is required in severe deformities. PMID- 11884119 TI - Reverse segmental pedicled ulna transfer as a salvage procedure in wrist fusion. AB - A new technique for wrist fusion using vascularized bone graft is described. A distally based, pedicled segment of the distal ulna, nourished by the ulnar artery or the distally based palmar-ulnar branch of the anterior interosseus artery was used in three patients to restore carpal height after infection (n=2) or tumour resection (n=1). The forearm is converted to a situation similar to a wide ulnar resection. All three wrist fusions healed uneventfully. This new technique is suitable in cases where a vascularized bone graft is required, but microsurgical techniques are not appropriate or are rejected by the patient. PMID- 11884120 TI - Penetrating injury to the terminal branches of the posterior interosseous nerve with nerve grafting. AB - We report two cases of penetrating injuries to the terminal branches of the posterior interosseous nerve in the forearm. Repair using nerve grafts in both cases were followed by complete recovery. PMID- 11884121 TI - Avulsion fracture of the extensor carpi radialis brevis insertion. AB - Avulsion of the extensor carpi radialis brevis at wrist level is rare. We present a case of an avulsion fracture involving the extensor carpi radialis brevis insertion at the base of the middle finger metacarpal. PMID- 11884123 TI - Should anatomic reduction be pursued in distal radial fractures. PMID- 11884122 TI - Re: neurophysiology not required before surgery for typical carpal tunnel syndrome. PMID- 11884124 TI - Re: closed reduction and percutaneous pinning of fractures of the proximal phalanx. PMID- 11884125 TI - Absorbable vs. non-absorbable sutures in carpal tunnel decompression. PMID- 11884126 TI - Carpal tunnel release under intravenous regional or local infiltration anaesthesia. PMID- 11884127 TI - The methyl group of N(alpha)(Me)Arg-containing peptides disturbs the active-site geometry of thrombin, impairing efficient cleavage. AB - Bivalent peptidic thrombin inhibitors consisting of an N-terminal d cyclohexylalanine-Pro-N(alpha)(Me)Arg active-site fragment, a flexible polyglycine linker, and a C-terminal hirugen-like segment directed towards the fibrinogen recognition exosite inhibit thrombin with K(i) values in the picomolar range, remaining stable in buffered solution at pH 7.8 for at least 15 hours. In order to investigate the structural basis of this increased stability, the most potent of these inhibitors, I-11 (K(i)=37pM), containing an N(alpha)(Me)Arg-Thr bond, was crystallized in complex with human alpha-thrombin. X-ray data were collected to 1.8A resolution and the crystal structure of this complex was determined. The Fourier map displays clear electron density for the N-terminal fragment and for the exosite binding segment. It indicates, however, that in agreement with Edman sequencing, the peptide had been cleaved in the crystal, presumably due to the long incubation time of 14 days needed for crystallization and data collection. The N(alpha)(Me) group is directed toward the carbonyl oxygen atom of Ser214, pushing the Ser195 O(gamma) atom out of its normal site. This structure suggests that upon thrombin binding, the scissile peptide bond of the intact peptide and the Ser195 O(gamma) are separated from each other, impairing the nucleophilic attack of the Ser195 O(gamma) toward the N(alpha)(Me)Arg carbonyl group. In the time-scale of two weeks, however, cleavage geometries favoured by the crystal allow catalysis at a slow rate. PMID- 11884128 TI - Transcriptional regulation by antitermination. Interaction of RNA with NusB protein and NusB/NusE protein complex of Escherichia coli. AB - A recombinant heterodimeric NusB/NusE protein complex of Escherichia coli was expressed under the control of a synthetic mini operon. Surface plasmon resonance measurements showed that the heterodimer complex has substantially higher affinity for the boxA RNA sequence motif of the ribosomal RNA (rrn) operons of E.coli as compared to monomeric NusB protein. Single base exchanges in boxA RNA reduced the affinity of the protein complex up to 15-fold. The impact of base exchanges in the boxA RNA on the interaction with NusB protein was studied by (1)H,(15)N heterocorrelation NMR spectroscopy. Spectra obtained with modified RNA sequences were analysed by a novel generic algorithm. Replacement of bases in the terminal segments of the boxA RNA motif caused minor chemical shift changes as compared to base exchanges in the central part of the dodecameric boxA motif. PMID- 11884129 TI - An SR-protein induced by HSVI binding to cells functioning as a splicing inhibitor of viral pre-mRNA. AB - The interaction between a virus and its specific receptor on the membrane of the host cell mimics the physiological combination of signal ligand and its receptor, and initiates the specific signal transduction from this activated receptor to induce a relative gene response. During the investigation of the interaction between Herpes simplex virus I (HSVI) and human fibroblast via the virus binding to its receptor complex on the cellular membrane, a new gene of cellular response against the specific stimulation of HSVI binding to fibroblasts was cloned from a cDNA library established from mRNA of an early gene response. This gene encoded a protein of 14.9kDa with the structural characteristics of Arg-rich and RS repeats. The analysis of the role of this protein in the infection by HSVI indicated that this protein, expressed only in G(1)/S phase and phosphorylated, functioned as a splicing inhibitor of HSVI pre-mRNA. The details of the mechanism of this inhibition of HSVI pre-mRNA splicing is still unclear. PMID- 11884130 TI - tRNA 3' end maturation in archaea has eukaryotic features: the RNase Z from Haloferax volcanii. AB - Here, we report the first characterization and partial purification of an archaeal tRNA 3' processing activity, the RNase Z from Haloferax volcanii. The activity identified here is an endonuclease, which cleaves tRNA precursors 3' to the discriminator. Thus tRNA 3' processing in archaea resembles the eukaryotic 3' processing pathway. The archaeal RNase Z has a KCl optimum at 5mM, which is in contrast to the intracellular KCl concentration being as high as 4M KCl. The archaeal RNase Z does process 5' extended and intron-containing pretRNAs but with a much lower efficiency than 5' matured, intronless pretRNAs. At least in vitro there is thus no defined order for 5' and 3' processing and splicing. A heterologous precursor tRNA is cleaved efficiently by the archaeal RNase Z. Experiments with precursors containing mutated tRNAs revealed that removal of the anticodon arm reduces cleavage efficiency only slightly, while removal of D and T arm reduces processing effciency drastically, even down to complete inhibition. Comparison with its nuclear and mitochondrial homologs revealed that the substrate specificity of the archaeal RNase Z is narrower than that of the nuclear RNase Z but broader than that of the mitochondrial RNase Z. PMID- 11884131 TI - Long-range correlations between DNA bending sites: relation to the structure and dynamics of nucleosomes. AB - It has been established that the precise positioning of nucleosomes on genomic DNA can be achieved, at least for a minority of them, through sequence-dependent processes. However, to what extent DNA sequences play a role in the positioning of the major part of nucleosomes is still debated. The aim of the present study is to examine to what extent long-range correlations (LRC) are related to the presence of nucleosomes. Using the wavelet transform technique, we perform a comparative analysis of the DNA text and of the corresponding bending profiles generated with curvature tables based on nucleosome positioning data. The exploration of a number of eukaryotic and bacterial genomes through the optics of the so-called "wavelet transform microscope" reveals a characteristic scale of 100-200 bp that separates two regimes of different LRC. Here, we focus on the existence of LRC in the small-scale regime (10-200 bp) which are actually observed in eukaryotic genomes, in contrast to their absence in eubacterial genomes. Analysis of viral DNA genomes shows that, like their host's genomes, eukaryotic viruses present LRC but eubacterial viruses do not. There is one exception for genomes of poxviruses (Vaccinia and Melamoplus sanguinipes) which do not replicate in the cell nucleus and do not exhibit LRC. No small-scale LRC are detected in the genomes of all examined RNA viruses, with the exception of retroviruses. These results together with the observation of LRC between particular sequence motifs known to participate in the formation of nucleosomes (e.g. AA dinucleotides) strongly suggest that the 10-200 bp LRC are a signature of the sequence-dependence of nucleosome positioning. Finally, we discuss possible interpretations of these LRC in terms of the physical mechanisms that might govern the positioning and the dynamics of the nucleosomes along the DNA chain through cooperative processes. PMID- 11884132 TI - Protein-splicing reaction via a thiazolidine intermediate: crystal structure of the VMA1-derived endonuclease bearing the N and C-terminal propeptides. AB - Protein splicing excises an internal intein segment from a protein precursor precisely, and concomitantly ligates flanking N and C-extein polypeptides at the respective sides of the precursor. Here, a series of precursor recombinants bearing 11 N-extein and ten C-extein residues is prepared for the intein of the Saccharomyces cerevisiae VMA1-derived homing endonuclease referred to as VDE and as PI-SceI. The recombinant with replacements of C284S, H362N, N737S, and C738S is chosen as a spliceable precursor model and is then subjected to a 2.1A resolution crystallographic analysis. The crystal structure shows that the introduced extein polypeptides are located in the vicinity of the splicing site, and that each of their peptide bonds is in the trans conformation. The S284 O(gamma) atom located at a distance of 3.1A from the G283 C atom in the N terminal junction suggests that a nucleophilic attack of the C284 S(gamma) atom on the G283 C atom forms a tetrahedral intermediate containing a five-membered thiazolidine ring. The tetrahedral intermediate is supposedly resolved into a thioester acyl group upon the cleavage of the linkage between the G283 C and C284 N atoms, and this thioester acyl formation completes the initial steps of Nright arrowS acyl shift at the junction between the N-extein and intein. The S738 O(gamma) atom in the C-terminal junction is placed in close proximity to the S284 O(gamma) atom at a distance of 3.6A, and is well suited for another nucleophilic attack on the resultant thioester acyl group that is then subjected to the transesterification in the next step. The reaction steps proposed for the acyl shift are driven entirely by protonation and deprotonation, in which proton ingress and egress is balanced within the splicing site. PMID- 11884133 TI - Crystal structures of a T4-lysozyme duplication-extension mutant demonstrate that the highly conserved beta-sheet region has low intrinsic folding propensity. AB - Residues 24 to 35 of T4 lysozyme correspond to the second and third strands of a region of beta-sheet that is highly conserved in all known lysozyme and chitinase structures. To evaluate the intrinsic propensity of these amino acid residues to form a defined structure they were added at the C terminus of the native protein, together with a dipeptide linker. Two crystal structures of this active, mutant protein were obtained, to 1.9A and 2.3A resolution, respectively. Even though the crystal conditions are similar, the appended sequence adopts very different secondary structures. In one case it is weakly structured and appears to extend through the active-site cleft, perhaps in part adding an extra strand to the original beta-sheet. In the other crystal form the extension is largely alpha helical. The formation of these alternative structures shows that the sequence does not have a strong intrinsic propensity to form a unique fold (either beta sheet or otherwise). The results also suggest that structural conservation during evolution does not necessarily depend on sequence conservation or the conservation of folding propensity. PMID- 11884134 TI - Artificial evolution of an enzyme active site: structural studies of three highly active mutants of Escherichia coli alkaline phosphatase. AB - The crystal structure of three mutants of Escherichia coli alkaline phosphatase with catalytic activity (k(cat)) enhancement as compare to the wild-type enzyme is described in different states. The biological aspects of this study have been reported elsewhere. The structure of the first mutant, D330N, which is threefold more active than the wild-type enzyme, was determined with phosphate in the active site, or with aluminium fluoride, which mimics the transition state. These structures reveal, in particular, that this first mutation does not alter the active site. The second mutant, D153H-D330N, is 17-fold more active than the wild type enzyme and activated by magnesium, but its activity drops after few days. The structure of this mutant was solved under four different conditions. The phosphate-free enzyme was studied in an inactivated form with zinc at site M3, or after activation by magnesium. The comparison of these two forms free of phosphate illustrates the mechanism of the magnesium activation of the catalytic serine residue. In the presence of magnesium, the structure was determined with phosphate, or aluminium fluoride. The drop in activity of the mutant D153H-D330N could be explained by the instability of the metal ion at M3. The analysis of this mutant helped in the design of the third mutant, D153G-D330N. This mutant is up to 40-fold more active than the wild-type enzyme, with a restored robustness of the enzyme stability. The structure is presented here with covalently bound phosphate in the active site, representing the first phosphoseryl intermediate of a highly active alkaline phosphatase. This study shows how structural analysis may help to progress in the improvement of an enzyme catalytic activity (k(cat)), and explains the structural events associated with this artificial evolution. PMID- 11884135 TI - Implications for the ubiquitination reaction of the anaphase-promoting complex from the crystal structure of the Doc1/Apc10 subunit. AB - The anaphase-promoting complex (APC) is a multi-subunit E3 protein ubiquitin ligase that is responsible for the metaphase to anaphase transition and the exit from mitosis. One of the subunits of the APC that is required for its ubiquitination activity is Doc1/Apc10, a protein composed of a Doc1 homology domain that has been identified in a number of diverse putative E3 ubiquitin ligases. Here, we present the crystal structure of Saccharomyces cerevisiae Doc1/Apc10 at 2.2A resolution. The Doc1 homology domain forms a beta-sandwich structure that is related in architecture to the galactose-binding domain of galactose oxidase, the coagulation factor C2 domain and a domain of XRCC1. Residues that are invariant amongst Doc1/Apc10 sequences, including a temperature sensitive mitotic arrest mutant, map to a beta-sheet region of the molecule, whose counterpart in galactose oxidase, the coagulation factor C2 domains and XRCC1, mediate bio-molecular interactions. This finding suggests the identification of the functionally important and conserved region of Doc1/Apc10 and, since invariant residues of Doc1/Apc10 colocalise with conserved residues of other Doc1 homology domains, we propose that the Doc1 homology domains perform common ubiquitination functions in the APC and other E3 ubiquitin ligases. PMID- 11884136 TI - The hidden thermodynamics of a zinc finger. AB - The Zn finger provides a model for studies of protein structure and stability. Its core contains a conserved phenylalanine residue adjoining three architectural elements: a beta-hairpin, an alpha-helix and a tetrahedral Zn(2+)-binding site. Here, we demonstrate that the consensus Phe is not required for high-affinity Zn(2+) binding but contributes to the specification of a precise DNA-binding surface. Substitution of Phe by leucine in a ZFY peptide permits Zn(2+)-dependent folding. Although a native-like structure is retained, structural fluctuations lead to attenuation of selected nuclear Overhauser enhancements and accelerated amide proton exchange. Surprisingly, wild-type Zn affinity is maintained by entropy-enthalpy compensation (EEC): a hidden entropy penalty (TDeltaDeltaS 7kcal/mol) is balanced by enhanced enthalpy of association (DeltaDeltaH 7kcal/mol) at 25 degrees C. Because the variant is less well ordered than the Phe anchored domain, the net change in entropy is opposite to the apparent change in configurational entropy. By analogy to the thermodynamics of organometallic complexation, we propose that EEC arises from differences in solvent reorganization. Exclusion of Leu among biological sequences suggests an evolutionary constraint on the dynamics of a Zn finger. PMID- 11884137 TI - The unusually slow relaxation kinetics of the folding-unfolding of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus. AB - In order to understand the thermodynamic and kinetic basis of the intrinsic stability of proteins from hyperthermophiles, the folding-unfolding reactions of cysteine-free pyrrolidone carboxyl peptidase (Cys142/188Ser) (PCP-0SH) from Pyrococcus furiosus were examined using circular dichroism (CD) and differential scanning calorimetry (DSC) at pH 2.3, where PCP-0SH exists in monomeric form. DSC showed a strong dependence of the shape and position of the unfolding profiles on the scan rate, suggesting the stability of PCP-0SH under kinetic control. On DSC timescales, even at a scan rate of 1 deg. C/hour, heat denaturation of PCP-0SH was non-equilibrium. However, over a long period of incubation of the heat denatured PCP-0SH at pre-transition temperatures, it refolded completely, indicating reversibility with very slow relaxation kinetics. The rates of refolding of the heat-denatured PCP-0SH determined from the time-resolved DSC and CD spectroscopic progress curves were found to be similar within experimental error, confirming the mechanism of refolding to be a two-state process. The equilibrium established with a relaxation time of 5080 seconds (at t(m)=46.5 degrees C), which is unusually higher than the relaxation times observed for mesophilic and hyperthermophilic proteins. The long relaxation time may lead to the apparent irreversibility of an unfolding process occurring on the DSC experiment timescale. The refolding rate (9.8 x 10(-5) s(-1)) peaked near the t(m) (=46.5 degrees C), whereas the stability profile reached maxima (11.8 kJ mol(-1)) at 17 degrees C. The results clearly indicate the unusual mode of protein destabilization via a drastic decrease in the rate of folding at low pH and still maintaining a high activation energy barrier (284 kJ mol(-1)) for unfolding, which provides an effective kinetic advantage to unusually stable proteins from hyperthermophiles. PMID- 11884139 TI - Preferential binding sites for interferon regulatory factors 3 and 7 involved in interferon-A gene transcription. AB - Transcription of the murine interferon-A4 (IFN-A4) gene is mediated by a virus responsive element (VRE-A4) located in the promoter proximal [-120 to -43] region. VRE-A4 contains four DNA modules (A to D) which cooperate for maximal IFN A4 activation following virus infection. The differential expression between the highly expressed IFN-A4 and the weakly inducible IFN-A11 gene promoters is essentially due to point mutations within the C and D modules of the virus responsive element VRE-A11. We now demonstrate that in murine L929 and human 293 cells, transcription factors IRF-3 and IRF-7, which are potent activators of virus-induced type I IFN transcription, differentially affect IFN-A4 and IFN-A11 promoter activities. Using electrophoretic mobility shift assays and DNase I footprinting data, our studies demonstrate that the AB modules correspond to a preferential site for IRF-7, whereas the C module is preferentially recognized by IRF-3. Furthermore, transfection of reporter constructs driven by four copies of different GAAANN hexameric motifs found within VRE-A4 indicates that the NN residues of these hexameric sequences define the preferential binding sites for IRF-3 or IRF-7. Together, these experiments clarify the molecular basis for differential expression of IFN-A genes following virus infection by delineating the sequence requirements for IRF association with the virus responsive elements of the IFN-A genes. PMID- 11884140 TI - p53 blocks RuvAB promoted branch migration and modulates resolution of Holliday junctions by RuvC. AB - The Holliday junction is the central intermediate in homologous recombination. Branch migration of this four-stranded DNA structure is a key step in genetic recombination that affects the extent of genetic information exchanged between two parental DNA molecules. Here, we have constructed synthetic Holliday junctions to test the effects of p53 on both spontaneous and RuvAB promoted branch migration as well as the effect on resolution of the junction by RuvC. We demonstrate that p53 blocks branch migration, and that cleavage of the Holliday junction by RuvC is modulated by p53. These findings suggest that p53 can block branch migration promoted by proteins such as RuvAB and modulate the cleavage by Holliday junction resolution proteins such as RuvC. These results suggest that p53 could have similar effects on eukaryotic homologues of RuvABC and thus have a direct role in recombinational DNA repair. PMID- 11884141 TI - Non-traditional Alu evolution and primate genomic diversity. AB - Alu elements belonging to the previously identified "young" subfamilies are thought to have inserted in the human genome after the divergence of humans from non-human primates and therefore should not be present in non-human primate genomes. Polymerase chain reaction (PCR) based screening of over 500 Alu insertion loci resulted in the recovery of a few "young" Alu elements that also resided at orthologous positions in non-human primate genomes. Sequence analysis demonstrated these "young" Alu insertions represented gene conversion events of pre-existing ancient Alu elements or independent parallel insertions of older Alu elements in the same genomic region. The level of gene conversion between Alu elements suggests that it may have a significant influence on the single nucleotide diversity within the genome. All the instances of multiple independent Alu insertions within the same small genomic regions were recovered from the owl monkey genome, indicating a higher Alu amplification rate in owl monkeys relative to many other primates. This study suggests that the majority of Alu insertions in primate genomes are the products of unique evolutionary events. PMID- 11884142 TI - Gene duplication and gene conversion shape the evolution of archaeal chaperonins. AB - Chaperonins are multi-subunit double-ring complexes that mediate the folding of nascent or denatured proteins. Gene duplication has been a potent force in the evolution of chaperonins in Archaea. Here we show that gene conversion has also been an important factor. We utilized a novel maximum likehood-based phylogenetic method for scanning DNA sequence alignments for regions of anomalous phylogenetic signal, such as those affected by gene conversion. Our results suggest that in crenarchaeotes, where an ancient gene duplication producing alpha and beta subunits took place in the common ancestor of the Pyrodictium, Aeropyrum, Pyrobaculum and Sulfolobus lineages, multiple independent gene conversions have occurred between the alpha and beta genes independently in each of these groups. Significantly, the conversions have repeatedly homogenized the region of the gene encoding the substrate-binding domain. This suggests that while the alpha and beta subunits in crenarchaeotes share only 50-60% overall amino acid sequence identity, they do not possess distinct roles in the binding of substrate. Cryptic gene conversion between distantly related paralogs may be more common than is currently appreciated, and could be a significant factor in slowing the functional differentiation of proteins encoded by duplicate genes long after their duplication. PMID- 11884144 TI - Atomic (0.94 A) resolution structure of an inverting glycosidase in complex with substrate. AB - The crystal structure of Clostridium thermocellum endoglucanase CelA in complex with cellopentaose has been determined at 0.94 A resolution. The oligosaccharide occupies six D-glucosyl-binding subsites, three on either side of the scissile glycosidic linkage. The substrate and product of the reaction occupy different positions at the reducing end of the cleft, where an extended array of hydrogen bonding interactions with water molecules fosters the departure of the leaving group. Severe torsional strain upon the bound substrate forces a distorted boat(2,5) B conformation for the glucosyl residue bound at subsite -1, which facilitates the formation of an oxocarbenium ion intermediate and might favor the breakage of the sugar ring concomitant with catalysis. PMID- 11884143 TI - Crystal structure of AlgQ2, a macromolecule (alginate)-binding protein of Sphingomonas sp. A1 at 2.0A resolution. AB - Sphingomonas sp. A1 possesses a high molecular mass (average 25,700 Da) alginate uptake system mediated by a novel pit-dependent ABC transporter. The X-ray crystallographic structure of AlgQ2 (57,200 Da), an alginate-binding protein in the system, was determined by the multiple isomorphous replacement method and refined at 2.0 A resolution with a final R-factor of 18.3% for 15 to 2.0 A resolution data. The refined structure of AlgQ2 was comprised of 492 amino acid residues, 172 water molecules, and one calcium ion. AlgQ2 was composed of two globular domains with a deep cleft between them, which is expected to be the alginate-binding site. The overall structure is basically similar to that of maltose/maltodextrin-binding protein, except for the presence of an N2-subdomain. The entire calcium ion-binding site is similar to the site in the EF-hand motif, but comprises a ten residue loop. This calcium ion-binding site is about 40 A away from the alginate-binding site. PMID- 11884145 TI - Mechanistic implications for Escherichia coli cofactor-dependent phosphoglycerate mutase based on the high-resolution crystal structure of a vanadate complex. AB - The structure of Escherichia coli cofactor-dependent phosphoglycerate mutase (dPGM), complexed with the potent inhibitor vanadate, has been determined to a resolution of 1.30 A (R-factor 0.159; R-free 0.213). The inhibitor is present in the active site, principally as divanadate, but with evidence of additional vanadate moieties at either end, and representing a different binding mode to that observed in the structural homologue prostatic acid phosphatase. The analysis reveals the enzyme-ligand interactions involved in inhibition of the mutase activity by vanadate and identifies a water molecule, observed in the native E.coli dPGM structure which, once activated by vanadate, may dephosphorylate the active protein. Rather than reflecting the active conformation previously observed for E.coli dPGM, the inhibited protein's conformation resembles that of the inactive dephosphorylated Saccharomyces cerevisiae dPGM. The provision of a high-resolution structure of both active and inactive forms of dPGM from a single organism, in conjunction with computational modelling of substrate molecules in the active site provides insight into the binding of substrates and the specific interactions necessary for three different activities, mutase, synthase and phosphatase, within a single active site. The sequence similarity of E.coli and human dPGMs allows us to correlate structure with clinical pathology. PMID- 11884146 TI - Differences in backbone dynamics of two homologous bacterial albumin-binding modules: implications for binding specificity and bacterial adaptation. AB - Proteins G and PAB are bacterial albumin-binding proteins expressed at the surface of group C and G streptococci and Peptostreptococcus magnus, respectively. Repeated albumin-binding domains, known as GA modules, are found in both proteins. The third GA module of protein G from the group G streptococcal strain G148 (G148-GA3) and the second GA module of protein PAB from P.magnus strain ALB8 (ALB8-GA) exhibit 59% sequence identity and both fold to form three helix bundle structures that are very stable against thermal denaturation. ALB8 GA binds human serum albumin with higher affinity than G148-GA3, but G148-GA3 shows substantially broader albumin-binding specificity than ALB8-GA. The (15)N nuclear magnetic resonance spin relaxation measurements reported here, show that the two GA modules exhibit mobility on the picosecond-nanosecond time scale in directly corresponding regions (loops and termini). Most residues in G148-GA3 were seen to be involved in conformational exchange processes on the microsecond millisecond time scale, whereas for ALB8-GA such motions were only identified for the beginning of helix 2 and its preceding loop. Furthermore, and more importantly, hydrogen-deuterium exchange and saturation transfer experiments reveal large differences between the two GA modules with respect to motions on the second-hour time scale. The high degree of similarity between the two GA modules with respect to sequence, structure and stability, and the observed differences in dynamics, binding affinity and binding specificity to different albumins, suggest a distinct correlation between dynamics, binding affinity and binding specificity. Finally, it is noteworthy in this context that the module G148-GA3, which has broad albumin-binding specificity, is expressed by group C and G streptococci known to infect all mammalian species, whereas P.magnus with the ALB8-GA module has been isolated only from humans. PMID- 11884147 TI - Structure, dynamics and binding characteristics of the second PDZ domain of PTP BL. AB - The PDZ domains of the protein tyrosine phosphatase PTP-BL mediate interactions by binding to specific amino acid sequences in target proteins. The solution structure of the second PDZ domain of PTP-BL, PDZ2, displays a compact fold with six beta strands and two alpha-helices. A unique feature of this domain compared to the canonical PDZ fold is an extended flexible loop at the base of the binding pocket, termed L1, that folds back onto the protein backbone, a feature that is shared by both the murine and human orthologues. The structure of PDZ2 differs significantly from the orthologous human structure. A comparison of structural quality indicators clearly demonstrates that the PDZ2 ensemble is statistically more reasonable than that of the human orthologue. The analysis of (15)N relaxation data for PDZ2 shows a normal pattern, with more rigid secondary structures and more flexible loop structures. Close to the binding pocket, Leu85 and Thr88 display greater mobility when compared to surrounding residues. Peptide binding studies demonstrated a lack of interaction between murine PDZ2 and the C terminus of the murine Fas/CD95 receptor, suggesting that the Fas/CD95 receptor is not an in vivo target for PDZ2. In addition, PDZ2 specifically binds the C termini of both human Fas/CD95 receptor and the RIL protein, despite RIL containing a non-canonical PDZ-interacting sequence of E-x-V. A model of PDZ2 with the RIL peptide reveals that the PDZ2 binding pocket is able to accommodate the bulkier side-chain of glutamic acid while maintaining crucial protein to peptide hydrogen bond interactions. PMID- 11884148 TI - Amino acid determinants of beta-hairpin conformation in erythropoeitin receptor agonist peptides derived from a phage display library. AB - Display of peptide libraries on filamentous phage has led to the identification of peptides of the form X(2-5)CX(2)GPXTWXCX(2-5) (where X is a variable residue) that bind to the extra-cellular portion of the erythropoietin receptor (EPO-R). These peptides adopt beta-hairpin conformations when co-crystallized with EPO-R. Solution NMR studies reveal that the peptide is conformationally heterogeneous in the absence of receptor due to cis-trans isomerization about the Gly-Pro peptide bond. Replacement of the conserved threonine residue with glycine at the turn i+3 position produces a stable beta-hairpin conformation in solution, although this peptide no longer has activity in an EPO-R-dependent cell proliferation assay. A truncated form of the EPO-R-binding peptide (containing the i+3 glycine residue) also forms a highly populated, monomeric beta-hairpin. In contrast, phage-derived peptide antagonists of insulin-like growth factor binding protein 1 (IGFBP-1) have a high level of sequence identity with the truncated EPO-R peptide (eight of 12 residues) yet adopt a turn-alpha-helix conformation in solution. Peptides containing all possible pairwise amino acid substitutions between the EPO-R and IGFBP-1 peptides have been analyzed to assess the degree to which the non conserved residues stabilize the hairpin or helix conformation. All four residues present in the original sequence are required for maximum population of either the beta-hairpin or alpha-helix conformation, although some substitutions have a more dominant effect. The results demonstrate that, within a given sequence, the observed conformation can be dictated by a small subset of the residues (in this case four out of 12). PMID- 11884150 TI - Conceptual models for implementing biopsychosocial theory in clinical practice. AB - The integration of the biopsychosocial model into manual therapy practice is challenging for clinicians, especially for those who have not received formal training in biopsychosocial theory or its application. In this masterclass two contemporary models of health and disability are presented along with a model for organizing clinical knowledge, and a model of reasoning strategies that will assist clinicians in their understanding and application of biopsychosocial theory. All four models emphasise the importance of understanding and managing both the psychosocial and the biomedical aspects of patients' problems. Facilitating change in patients' (and clinicians') perspectives on pain and its biopsychosocial influences requires them to reflect on their underlying assumptions and the basis of those beliefs. Through this reflective process perspectives will be transformed, and for clinicians, in time, different management practices will emerge. PMID- 11884151 TI - A survey to examine attitudes and patterns of practice of physiotherapists who perform cervical spine manipulation. AB - As part of the process of developing a Clinical Practice Guideline (CPG) on cervical spine manipulation (CSM), a working group sent out an 82-item postal survey to 150 randomly selected Ontario physiotherapists (PTs) who perform spinal manipulation, to collect information on the socio-demographics, practices, opinions of risk, and attitudes towards CPGs of these PTs (n = 118; response rate = 79%). Of the 118 respondents who performed spinal manipulation, 41 performed CSM. Respondents strongly agreed with three out of six indications listed in the survey for applying CSM: segmental fixation, stiff but stable joint, internal derangement (over 70%). Respondents also strongly agreed (over 88%) that all screening tests listed in the survey should be performed prior to applying CSM: tests for irritability, stability, vascular and neurological systems. Respondents rated patient education, other manual therapy, and exercise as the most common adjuncts to CSM (over 88%). Respondents reported seeing mild complications or side effects only rarely following the application of CSM. Fourteen percent of respondents reported having a written CSM policy or CPG on CSM in their work setting. Feedback from this survey will be used in developing a CSM CPG. A future survey will evaluate changes in clinical practice and in attitudes toward CPGs some time after the dissemination of the CSM CPG. PMID- 11884152 TI - The response of posteroanterior lumbar stiffness to repeated loading. AB - Lumbar posteroanterior (PA) responses are determined by manual examination and are used to guide treatment decisions and interpret changes in symptoms within and between treatments. Mechanical devices that simulate manual assessment have been developed to measure lumbar PA responses. The two variables used to describe lumbar PA responses to mechanical loading are stiffness coefficient K and displacement D30. The purpose of this study was to investigate the behaviour of lumbar PA responses with repeated loading over time. Lumbar PA responses at L4 were measured in 18 pain-free subjects using a mechanical device. Measurements were made for five consecutive loading cycles on three test occasions. The responses were compared between the five cycles within a single test occasion and between three test occasions. An identical procedure was also used to test a set of elastic springs for comparison. There was a significant increase in both stiffness coefficient K and displacement D30 between the first cycle and subsequent cycles of a single test occasion on human subjects. This response which demonstrates an increase in stiffness and displacement between the first and subsequent cycles can be considered a normal response to PA loading. PA stiffness remains constant over several tests both within one day and between days. PMID- 11884153 TI - Scapular position: the validity of skin surface palpation. AB - The assessment of the resting position of the scapula forms part of the examination of upper quadrant posture. The purpose of this study was to determine if surface palpation is a valid indicator of actual scapular position. Twelve embalmed shoulders were examined and the actual location of three bony scapular landmarks and three bony thoracic landmarks were compared with surface palpation of these locations. The results, based on the upper value for a distance (with 95% confidence), suggested that the difference between the surface location of the root of the spine of the scapula, the acromial angle and the inferior angle would be less than 0.67 cm, 0.98 cm and 0.46 cm respectively from the centre of the bony locations. The difference between the twelfth thoracic spinous process (SP), the SP corresponding with the root of the spine of the scapula, and the SP corresponding with the inferior angle and the surface points would be 1.46 cm, 1.09 cm and 1.01 cm respectively. The results of this study suggest that surface palpation of scapular location is a valid method for determining the actual location of the scapula. The findings also suggest that surface palpation can determine the location of thoracic landmarks, which may serve as reference points for scapular position. PMID- 11884154 TI - Effectiveness of manipulative physiotherapy for the treatment of a neurogenic cervicobrachial pain syndrome: a single case study -- experimental design. AB - A single case study ABC design was used to evaluate the effectiveness of manipulative physiotherapy in a 44-year-old woman with an 8-month history of neurogenic cervicobrachial pain. Clinical examination demonstrated significant signs of upper quadrant neural tissue mechanosensitivity indicating that neural tissue was the dominant tissue of origin for the subject's complaint of pain. Magnetic resonance imaging revealed correlating discal pathology at the C5/6 intersegmental level. The study involved a 4-week pre-assessment phase, a 4-week treatment phase and a 2-week home exercise phase. Functional disability was measured using the Northwick Park Neck Pain Questionnaire and pain was assessed using the McGill Short Form Pain Questionnaire. Cervical motion was measured by a cervical range of motion device (CROM) and the range of shoulder abduction with a mediclino inclinometer. Manipulative physiotherapy treatment involved a cervical lateral glide mobilization technique. Following treatment, visual analysis revealed beneficial effects on pain, functional disability as well as cervical and shoulder mobility. These improvements were maintained over the home exercise phase and at 1-month follow-up. The single case limits generalization of the findings, but the results support previous studies in this area and gives further impetus to controlled clinical trials. PMID- 11884155 TI - Manual examination of accessory movements--seeking R1. AB - Movement diagrams are used by physiotherapists to depict the behaviour of resistance through the available range of accessory and physiological joint movement. It is generally accepted that for an asymptomatic joint, the resistance first felt by the therapist (R1) occurs towards the end of range. R1 is considered to be at the transition point between the toe and linear region of a load displacement curve. The aim of this study was to more accurately define R1 from force displacement curves of accessory movement to the spine and peripheral joints using a validated instrument, the Spinal Assessment Machine (SAM). Thirty archived force displacement curves obtained using the SAM, which applied a posteroanterior force of 100N at a frequency of 0.5 Hz to L3 spinous process, were examined. In addition force displacement curves were similarly obtained from the tibiofemoral joint, glenohumeral joint and radiocarpal joint of one asymptomatic individual. In all cases resistance to a PA movement commenced at the beginning of range, the curve ascending as soon as the force was applied. While in most cases there was a low stiffness 'toe' region there was no unambiguous point where it could be said that the toe region ended. It is concluded that for spinal and peripheral accessory movements both the onset of resistance and the toe occurs at the beginning of range. Therapists should therefore depict R1 at the beginning of range not toward the end of range as is current practice. PMID- 11884156 TI - Mulligan's mobilization with movement for the thumb: a single case report using magnetic resonance imaging to evaluate the positional fault hypothesis. PMID- 11884157 TI - Bibliography. Cartilage, Articular. PMID- 11884159 TI - A phylogenetic comparison of red deer and wapiti using mitochondrial DNA. AB - A phylogeny was constructed for red deer/wapiti (Cervus elaphus) subspecies using sequence data from the control region of mitochondrial DNA (mtDNA). The tree was rooted using Cervus nippon (sika deer), Cervus albirostris (Thorold's white lipped deer), and several Odocoileinae species. A division between the mtDNA haplotypes of red deer (European) and wapiti (Asian/North American) corresponds to subspecies found on opposite sides of the Himalayan Mountains and Gobi, which suggests wapiti should be reconsidered for the status of C. canadensis. Using parsimony and distance analysis, red deer and wapiti are derived from a single recent common ancestor, which is consistent with current taxonomy that recognizes the subspecies of Cervus elaphus as monophyletic group. However, maximum likelihood analysis using weighted transitional substitutions caused red deer to form a sister group to sika deer (Cervus nippon) and wapiti. A phenetic comparison revealed wapiti also share more nucleotide similarities with sika deer, although approximately 5% sequence divergence separates wapiti, sika, and red deer. Phylogenetic evidence from the cytochrome b sequences corroborated observations from the control region. Observations from this study suggest that the species status of wapiti should be reinstated. PMID- 11884158 TI - Characterization and phylogenetic utility of the mammalian protamine p1 gene. AB - We sequenced the protamine P1 gene (ca. 450 bp) from 20 bats (order Chiroptera) and the flying lemur (order Dermoptera). We compared these sequences with published sequences from 19 other mammals representing seven orders (Artiodactyla, Carnivora, Cetacea, Perissodactyla, Primates, Proboscidea, and Rodentia) to assess structure, base compositional bias, and phylogenetic utility. Approximately 80% of second codon positions were guanine, resulting in protamine proteins containing a high frequency of arginine residues. Our data indicate that codon usage for arginine differs among higher mammalian taxa. Parsimony analysis of 40 species representing nine orders produced a well-resolved tree in which most nodes were supported strongly, except at the lowest taxonomic levels (e.g., within Artiodactyla and Vespertilionidae). These data support monophyly of several taxa proposed by morphologic and molecular studies (all nine orders: Laurasiatheria, Cetartiodactytla, Yangochiroptera, Noctilionoidea, Rhinolophoidea, Vespertilionoidea, Phyllostomidae, Natalidae, and Vespertilionidae) and, in agreement with recent molecular studies, reject monophyly of Archonta, Volitantia, and Microchiroptera. Bats were sister to a clade containing Perissodactyla, Carnivora, and Cetartiodactyla, and, although not unequivocally, rhinolophoid bats (traditional microchiropterans) were sister to megachiropterans. Sequences of the protamine P1 gene are useful for resolving relationships at and above the familial level in bats, and generally within and among mammalian orders, but with some drawbacks. The coding and intervening sequences are small, producing few phylogenetically informative characters, and aligning the intron is difficult, even among closely related families. Given these caveats, the protamine P1 gene may be important to future systematic studies because its functional and evolutionary constraints differ from other genes currently used in systematic studies. PMID- 11884160 TI - Molecular evidence for the monophyly of tenrecidae (mammalia) and the timing of the colonization of Madagascar by Malagasy Tenrecs. AB - Tenrecs are a diverse family of insectivores, with an Afro-Malagasian biogeographic distribution. Three subfamilies (Geogalinae, Oryzorictinae, Tenrecinae) are restricted to Madagascar and one subfamily, the otter shrews (Potamogalinae), occurs on the mainland. Morphological studies have generated conflicting hypotheses according to which both tenrecids and Malagassy tenrecs are either monophyletic or paraphyletic. Competing hypotheses have different implications for the biogeographic history of Tenrecidae. At present, there are no molecular studies that address these hypotheses. The present study provides sequences of a nuclear protein-coding gene (vWF) and the mitochondrial 12S rRNA, tRNA valine, and 16S rRNA genes from a potamogaline (Micropotamogale). New sequences of these genes are also reported for the tenrecine, Tenrec ecaudatus. The 12S sequences from these taxa were combined with data already available for this locus from two other tenrecids (Echinops telfairi, subfamily Tenrecinae and Oryzorictes talpoides, subfamily Oryzorictinae). Phylogenetic analyses provided strong bootstrap support for the monophyly of Tenrecidae and Malagasy tenrecs. The majority of statistical tests rejected morphological claims for both a Tenrecinae--Chrysochloridae clade and an Oryzorictinae--Potamogalinae clade. Molecular clock estimates suggest a split of otter shrews and Malagasy tenrecs at approximately 53 MYA. We estimate that the ancestor of Malagasy tenrecs dispersed to Madagascar subsequent to this split but prior to about 37 MYA. PMID- 11884161 TI - Molecular phylogeny of the snubnose darters, subgenus Ulocentra (genus Etheostoma, family Percidae). AB - Snubnose darters comprise one of the largest subgenera of the percid genus Etheostoma. Many species are described based on differences in male breeding coloration. Few morphological synapomorphies have been proposed for the subgenus and their relatives, making it difficult to delineate monophyletic clades. The phylogenetic relationships of the 20 snubnose darter species of the subgenus Ulocentra and 11 members of its proposed sister subgenus Etheostoma were investigated with partial mitochondrial DNA sequences including 1033 bp encompassing the entire mitochondrial control region, the tRNA-Phe gene, and part of the 12S rRNA gene. Two hypotheses on the relationship and monophyly of the two subgenera were evaluated. Both maximum-parsimony and neighbor-joining analyses supported monophyly of the subgenus Ulocentra and resolved some species-level relationships. The banded darter, E. zonale, and its sister taxon, E. lynceum, were not closely related to the snubnose darters and appear to be diverged from the other members of the subgenus Etheostoma, fitting their former distinction as the recognized subgenus Nanostoma. The sister group to Ulocentra appears to be a restricted species assemblage within the subgenus Etheostoma containing E. blennioides, E. rupestre, E. blennius, and the E. thalassinum species group. The placement of the harlequin darter, E. histrio, is problematic, and it may represent a basal member of Ulocentra or of the restricted subgenus Etheostoma. Despite recent estimates of divergence times between nominal Ulocentra taxa, each species exhibits its own unique set of mtDNA haplotypes, providing no direct evidence for current genetic exchange between species. The nominal taxa of snubnose darters thus appear to be evolving independently from each other and therefore constitute valid species under the Phylogenetic Species Concept. PMID- 11884162 TI - Mitochondrial phylogeography of moose (Alces alces): late pleistocene divergence and population expansion. AB - We examined phylogeographic relationships of moose (Alces alces) worldwide to test the proposed existence of two geographic races and to infer the timing and extent of demographic processes underpinning the expansion of this species across the Northern Hemisphere in the late Pleistocene. Sequence variation within the left hypervariable domain of the control region occurred at low or moderate levels worldwide and was structured geographically. Partitioning of genetic variance among regions indicated that isolation by distance was the primary agent for differentiation of moose populations but does not support the existence of distinct eastern and western races. Levels of genetic variation and structure of phylogenetic trees identify Asia as the origin of all extant mitochondrial lineages. A recent coalescence is indicated, with the most recent common ancestor dating to the last ice age. Moose have undergone two episodes of population expansion, likely corresponding to the final interstade of the most recent ice age and the onset of the current interglacial. Timing of expansion for the population in the Yakutia--Manchuria region of eastern Asia indicates that it is one of the oldest populations of moose and may represent the source of founders of extant populations in North America, which were colonized within the last 15,000 years. Our data suggest an extended period of low population size or a severe bottleneck prior to the divergence and expansion of extant lineages and a recent, less-severe bottleneck among European lineages. Climate change during the last ice age, acting through contraction and expansion of moose habitat and the flooding of the Bering land bridge, undoubtedly was a key factor influencing the divergence and expansion of moose populations. PMID- 11884163 TI - Molecular phylogenetics of emydine turtles: taxonomic revision and the evolution of shell kinesis. AB - The 10 extant species of emydine turtles represent an array of morphological and ecological forms recognizable and popular among scientists and hobbyists. Nevertheless, the phylogenetic affinities of most emydines remain contentious. Here, we examine the evolutionary relationships of emydine turtles using 2092 bp of DNA encoding the mitochondrial genes cyt b, ND4, and adjacent tRNAs. These data contain 339 parsimony informative characters that we use to erect hypotheses of relationships for the Emydinae. Both maximum parsimony and maximum likelihood methods yield a monophyletic Emydinae in which all but three nodes are well resolved. Emys orbicularis, Emydoidea blandingii, and Clemmys marmorata form a monophyletic clade, as do the species of Terrapene. Clemmys muhlenbergii and Clemmys insculpta form a third monophyletic group that may be sister to all other emydines. Clemmys guttata is problematic and probably related to Terrapene. Based on this phylogeny, and previous molecular work on the group, we suggest the following taxonomic revisions: (1) Clemmys should be restricted to a single species, C. guttata. (2) Calemys should be resurrected for C. muhlenbergii and C. insculpta. (3) Emys should be expanded to include three species: E. orbicularis, E. blandingii, and E. marmorata. Furthermore, our analyses show that neither kinetic-shelled nor akinetic-shelled emydines form monophyletic groups. Therefore, shell kinesis was either independently gained in Emys and Terrapene or secondarily lost in E. marmorata and C. guttata. Parsimony, paleontological evidence, and the multiple origins of shell kinesis in related turtle lineages (especially geoemydines) support the independent origin of plastral kinesis. PMID- 11884164 TI - Molecular phylogeny of a circum-global, diverse gastropod superfamily (Cerithioidea: Mollusca: Caenogastropoda): pushing the deepest phylogenetic limits of mitochondrial LSU rDNA sequences. AB - The Cerithioidea is a very diverse group of gastropods with ca. 14 extant families and more than 200 genera occupying, and often dominating, marine, estuarine, and freshwater habitats. While the composition of Cerithioidea is now better understood due to recent anatomical and ultrastructural studies, the phylogenetic relationships among families remain chaotic. Morphology-based studies have provided conflicting views of relationships among families. We generated a phylogeny of cerithioideans based on mitochondrial large subunit rRNA and flanking tRNA gene sequences (total aligned data set 1873 bp). Nucleotide evidence and the presence of a unique pair of tRNA genes (i.e., threonine + glycine) between valine-mtLSU and the mtSSU rRNA gene support conclusions based on ultrastructural data that Vermetidae and Campanilidae are not Cerithioidea, certain anatomical similarities being due to convergent evolution. The molecular phylogeny shows support for the monophyly of the marine families Cerithiidae [corrected], Turritellidae, Batillariidae, Potamididae, and Scaliolidae as currently recognized. The phylogenetic data reveal that freshwater taxa evolved on three separate occasions; however, all three recognized freshwater families (Pleuroceridae, Melanopsidae, and Thiaridae) are polyphyletic. Mitochondrial rDNA sequences provide valuable data for testing the monophyly of cerithioidean [corrected] families and relationships within families, but fail to provide strong evidence for resolving relationships among families. It appears that the deepest phylogenetic limits for resolving caenogastropod relationships is less than about 245--241 mya, based on estimates of divergence derived from the fossil record. PMID- 11884165 TI - A molecular phylogeny of the frog genus Tomopterna in Southern Africa: examining species boundaries with mitochondrial 12S rRNA sequence data. AB - Frogs of the genus Tomopterna occur throughout sub-Saharan Africa. Previous work has shown that there are seven cryptic species, which occupy diverse habitats from grasslands to deserts. The current paper proposes a phylogeny of Tomopterna based on partial sequences of the mitochondrial 12S rRNA gene. A gene tree for the genus, including all seven named species and three undescribed species which were discovered during the course of this study, is presented. PMID- 11884166 TI - 18S rDNA phylogeny of the Tubificidae (Clitellata) and its constituent taxa: dismissal of the Naididae. AB - The phylogeny of the Tubificidae, and of most of its subfamilies and some of its genera, is revisited, on the basis of sequences of 18S ribosomal DNA in a selection of species. Forty-six new 18S sequences of Naididae (6), Tubificidae (37), Phreodrilidae (1), Lumbriculidae (1), and Enchytraeidae (1) are reported and aligned together with corresponding sequences of 21 previously studied taxa. The 18S gene of Insulodrilus bifidus provides the first molecular evidence that phreodrilids are closely related to tubificids, corroborating previous conclusions based on morphology. The data further support the monophyletic status of Tubificidae, provided that the "Naididae" is regarded a part of this family; "naidids" may not even constitute a monophyletic group. It is thus suggested that the family name Naididae is formally suppressed as a junior synonym of the Tubificidae. The 18S gene also resolves a number of relationships within the tubificids. Among the subfamilies, Tubificinae is supported, Rhyacodrilinae and Phallodrilinae are revealed as nonmonophyletic, and Limnodriloidinae remains unresolved. Most tubificid genera tested for monophyly are corroborated by the data, only one (Tubifex) is refuted, and two (Tubificoides and Limnodriloides) are unresolved from other taxa. It is concluded that it will be valuable to expand the taxonomic sampling for 18S rDNA in clitellates, and in annelids in general, as this is likely to improve the resolution at many levels. However, it will be equally important to combine the annelid 18S data with other gene sequences and nonmolecular characters, to estimate the phylogeny of these common and diverse worms with greater precision. PMID- 11884167 TI - Phylogenetic relationships in Termitomyces (Family Agaricaceae) based on the nucleotide sequence of ITS: a first approach to elucidate the evolutionary history of the symbiosis between fungus-growing termites and their fungi. AB - Termitomyces constitutes a very poorly known genus of fungi whose essential characteristic is that all representatives of the genus are cultivated by termites (Macrotermitinae) in their nest and that all the fungi cultivated by termites belong to this genus. For the first time, the phylogenetic relationships of several African Termitomyces species was studied by the sequencing of their internal transcriber spacer region (ITS1--5.8S--ITS2). It appeared that this group is clearly monophyletic and belongs to the Tricholomataceae family. The total homology of the ITS zone of several Termitomyces symbionts of different termite genera indicated that the specific diversity of this group is in fact less important than previously supposed. Finally, the comparison between the Termitomyces phylogenetic tree and the taxonomic tree of Macrotermitinae showed that if for certain genera the hypothesis of termite/fungus coevolution is acceptable, it should not be applied for all symbiosis. PMID- 11884168 TI - Phylogeny of the tree swallow genus, Tachycineta (Aves: Hirundinidae), by Bayesian analysis of mitochondrial DNA sequences. AB - To set the stage for historical analyses of the ecology and behavior of tree swallows and their allies (genus Tachycineta), we reconstructed the phylogeny of the nine Tachycineta species by comparing DNA sequences of six mitochondrial genes: Cytochrome b (990 base pairs), the second subunit of nicotinamide adenine dinucleotide dehydrogenase (839 base pairs), cytochrome oxidase II (85 base pairs), ATPase 8 (158 base pairs), tRNA-lysine (73 base pairs), and tRNA methionine (25 base pairs). The phylogeny consisted of two main clades: South and Central American species ((T. stolzmanni, T. albilinea, T. albiventris), (T. leucorrhoa, T. meyeni)), and North American and Caribbean species (T. bicolor, (T. thalassina, T. euchrysea, T. cyaneoviridis)). The genetic distances among the species suggested that Tachycineta is a relatively old group compared to other New World swallow genera. One interesting biogeographic discovery was the close relationship between Caribbean and western North American taxa. This historical connection occurs in other groups of swallows and swifts as well. To reconstruct the phylogeny, we employed Bayesian as well as traditional maximum-likelihood methods. The Bayesian approach provided probability values for trees produced from the different genes and gene combinations, as well as probabilities of branches within those trees. We compared Bayesian and maximum-likelihood bootstrap branch support and found that all branches with Bayesian probabilities > or = 95% received bootstrap support >70%. PMID- 11884170 TI - Molecular relationships among European samples of Reticulitermes (Isoptera, Rhinotermitidae). AB - Taxonomy and phyletic relationships of the European termites of the holarctic genus Reticulitermes are highly debated and poorly known. Sequencing analyses of the complete cytochrome oxidase subunit II and of a fragment of 16S rDNA mitochondrial genes were performed on 21 Italian and French populations. Distance, parsimony, and maximum-likelihood evaluations on single and combined data sets confirm the presence in central peninsular Italy of R. lucifugus lucifugus, but demonstrate that a coastal Tuscan population and the Sardinian ones pertain to the Corsican subspecies R. lucifugus corsicus, whose distribution appears therefore transtyrrenian. Northeastern Italian samples are highly differentiated from presently analyzed R. lucifugus and constitute a new entity, possibly more related to the Japanese R. speratus. Finally, the French R. santonensis and the North American R. flavipes appear strongly related, so that the supposed synonymy between the two taxa must be accepted. PMID- 11884169 TI - Phylogeny of the subgenus sophophora (Diptera: drosophilidae) based on combined analysis of nuclear and mitochondrial sequences. AB - Sequences from the nuclear (nu) alcohol dehydrogenase gene, the nu 28S ribosomal RNA locus, and the mitochondrial cytochrome oxidase II gene were used both individually and in combined analyses to infer the phylogeny of the subgenus Sophophora (Diptera: Drosophilidae). We used several optimality criteria, including maximum likelihood, maximum parsimony, and minimum evolution, to analyze these partitions to test the monophyly of the subgenus Sophophora and its four largest species groups, melanogaster, obscura, saltans, and willistoni. Our results suggest that the melanogaster and obscura species groups are each monophyletic and form a closely related clade. The Neotropical clade, containing the saltans and willistoni species groups, is also recovered, as previous studies have suggested. While the saltans species group is strongly supported as monophyletic, the results of several analyses indicate that the willistoni species group may be paraphyletic with respect to the saltans species group. PMID- 11884171 TI - Nurse education in Ireland: redressing the balance. PMID- 11884172 TI - Accredited work-based learning: an approach for collaboration between higher education and practice. AB - This article discusses the experience of creating a programme of accredited work based learning (AWBL) for emergency nurse practitioners (ENPs) who work in an Accident and Emergency (A&E) Department in the North East of England. The initiative highlighted the challenges of collaboration with purchasers of education and with professional colleagues, other than nurses. Accredited work based learning was seen to be an appropriate means of supporting ENP role development. Some of the drivers of the development were: the need for a rapid response to wide ranging changes in the health service; the need to ensure rigour in the quality of both education and health care; to enable participants to engage in role development with appropriate skills, confidence and competence; and to ensure that the learning programme had parity in its design with conventional university based learning. The aim was to collaborate in the creation of learning which was relevant to the Trust's drive to improve patient care which accommodated the nurses' common and individual learning needs and offered academically recognized learning opportunities in tune with the post Dearing ethos in higher education. This aim was reached and included a great deal of learning on the part of the collaborating partners. PMID- 11884173 TI - Lifelong learning in nursing: perceptions and realities. AB - Because of ongoing development and changes in health care delivery, lifelong learning, as a concept and a practical activity, has increasingly gained centre stage in the nursing profession. There seems, however, to be no research study on how nurses feel about the introduction and implementation of this notion. Additionally, despite lifelong learning being advocated for all in UK society for a variety of reasons, there are other perceptions of the concept. This article reviews the literature to determine the meaning of the concept, which is then critically analysed in conjunction with aspects of qualitative data elicited from nurses directly involved in clinical nursing practice. Data from 26 semi structured individual interviews conducted as part of study for a Doctor in Philosophy (PhD) course exploring nurses' perceptions of lifelong learning and analysis of contemporary reports and documents indicate that although there are a number of very useful facets to the concept, there are also certain reservations directed at participation in lifelong learning. PMID- 11884174 TI - Re-introducing skills teaching to nurse education: an action research project. AB - A skills teaching programme for first year nursing students is being developed as an action research project with the participation of lecturers, practitioners of three NHS trusts and first year students. All branches of nursing are equally represented. During the first part of the action research cycle the skills to be taught were decided upon using anonymous questionnaires and branch specific group interviews. The selected skills related to aspects of communication, observation, care planning and nursing activities as well as health care and information technology. The professional and personal development of students was also considered. All branches were able to agree on a common skills teaching programme. During this first part of the action research cycle concerns about the organizational possibility of teaching skills to an ever-increasing number of nursing students and the necessary participation of many lecturers were raised. Nevertheless, all participants emphasized the need to continue the project, requesting the development, implementation and evaluation of a new curriculum based on this research as soon as possible. This research demonstrates how small schools of nursing without the resources of large metropolitan-based medical schools and teaching hospitals can improve or design and implement skills teaching programmes for their students in a logical and research based fashion. PMID- 11884175 TI - Development of an innovative tool to assess hospital learning environments. AB - Nursing as a profession has evolved in response to societal needs for well prepared practitioners who provide quality care to the needed in episodes of illness, and promote health among all age groups. Clinical practice enables the student to develop competencies in the application of knowledge, skills, and attitudes to clinical field situations. The clinical learning environment is a multidimensional entity with a complex social context. Previous research on clinical learning environment was examined, yet minimal studies have been conducted on hospital learning environments from the psychosocial educational perspective. With the mission to maximize nursing students' clinical learning experiences, the author developed and validated the Clinical Learning Environment Inventory (CLEI) based on the theoretical framework in psychosocial education. It is envisaged that this tool will assist the nurse professional to facilitate nursing students to achieve a productive clinical practice. This paper highlights the need and details the conceptual framework in the development of the CLEI. Although it is not presented as a research based paper, some statistical data are included to verify the reliability and validity of the newly-developed instrument. PMID- 11884176 TI - Facilitating the development of clinical skills in caring for dying people in hospital. AB - Research shows that diploma level preparation has resulted in newly qualified nurses with clinical skill deficits (e.g. Macleod Clark et al. 1996, Carlisle et al. 1999). Part of the government's current plan to improve the quality of services delivered by the NHS is to address this problem through the introduction of a new nursing role; a role incorporating both clinical practice and teaching (Department of Health 1999). This paper draws data from a phenomenological study of 28 qualified diplomat nurses. It identifies their self-perceived skill deficits in relation of caring for dying people in acute hospital medical wards, and the ways that they believe that they could best be helped to overcome these deficits. On the basis of these findings, strategies are recommended that a practitioner-educator might adopt, with the intention of facilitating skill development in the care of dying people. PMID- 11884177 TI - Empowering moral decision making in nurses. AB - In the past years a schooling programme in moral decision making has been provided in an Amsterdam acute care hospital for health care professionals. The goal has been to heighten awareness, stimulate communication between disciplines regarding moral dilemmas and support members of different disciplines to reach commonly shared decisions. This article discusses the outcomes of the schooling programme and shows that an investment of this type improves communication amongst disciplines and contributes towards the empowerment of nurses. PMID- 11884178 TI - Serving two masters: quality teaching and learning versus economic rationalism. AB - Nurse educators face the challenge of competing pressures. Programmes must be developed that more adequately prepare students to meet the demands of a changing and complex health care system. These programmes must reflect excellence in teaching and learning and this needs to be achieved within the constraints of economic rationalism. The design of a model based on principles of self directed learning assisted one university to deliver a high quality clinical skills programme. PMID- 11884179 TI - An education programme for professionals who specialize in substance misuse in St Petersburg, Russia: part 1. AB - This paper provides an account of a joint project of education and training of doctors and nurses in St Petersburg, funded by the Know How Fund Health Sector Small Partnership Scheme (Russia). Contextual material on drug and alcohol misuse in Russia is introduced prior to a focus on the drug and alcohol misuse issues in St Petersburg. Reference is made to historical and contemporary material on alcohol and drug misuse, and attention is drawn to the reliability of statistical data. The main aims of the project and the work carried out are outlined. Firstly, to bring together medical and nursing colleagues, enabling a recognition of the overlap in training and educational needs of both professional groups, and the learning that can occur from understanding each others roles and responsibilities. Secondly, that the theory and practice of different approaches to care and treatment can be incorporated into already established curricula used to educate both nurses and doctors. Colleagues in St Petersburg have requested the support and guidance of UK practitioners and nurse educationalists to facilitate these changes in perspectives. The paper concludes with a brief discussion of the influence of the project in anticipation of a forthcoming paper that will detail evaluation processes that the provision has undergone and examine the findings in more detail. PMID- 11884180 TI - User involvement in mental health branch education: client review presentations. AB - This paper will present the work of users, students and lecturers involved in the delivery of Mental Health pre-registration education, focusing on client assessment. Users from local mental health representative organizations, attend a series of college-based sessions to evaluate student's client review presentations, in which individual students explicitly and critically reflect upon a mental health assessment of a client in which they have participated. The user contributions are intended to raise student's awareness of client-centred perspectives, particularly in terms of the various possible interpretations that may be attributed to assessment data gathered about the client. The sessions seek to develop a learning approach, which will develop and consolidate a partnership in curriculum delivery between mental health service users and nursing education. This method of working has been evaluated by a previous study, and indicates that this method has an important influence on student's approaches to identifying clients needs and subsequent care delivery. This paper will focus on the organizational pre-requisites that are desirable, to implement this method of teaching and learning, including philosophical issues, contract arrangements, classroom activities, supervision, consideration of ethical dilemmas and reflective outcomes. PMID- 11884181 TI - Transcultural nursing education: a view from within. AB - There has been increasing interest in the nursing care of patients from minority ethnic communities in recent years. This has been linked to the promotion of transcultural competence within the profession in order to equip nurses to provide high quality and appropriate care within a multi-ethnic society. Nurse education has been identified as an ideal vehicle for the promotion of transcultural competence in both students and qualified nursing staff. However, many of the current approaches to transcultural education appear to reflect a single ethnocentric approach to transcultural education. This fails to appreciate the diversity which exists within both the profession and minority ethnic communities.The paper is written from a minority ethnic perspective and explores some of the issues for transcultural nurse education arising from this ethnocentric approach. It discusses some common approaches underpinning the provision of transcultural education and the potential consequences for a multi ethnic profession seeking to 'value diversity'. It concludes that a review of some of the current approaches to transcultural education is required and suggests some practical points for consideration. PMID- 11884187 TI - Information and communication technologies (ICT) in nursing education: is there a need for a more philosophical analysis? PMID- 11884188 TI - The Nurse Education Tomorrow conference: a critique. PMID- 11884189 TI - The Nurse Education Tomorrow conference: a critique. PMID- 11884190 TI - Courses for horses: is nursing a front runner in the higher education stakes? PMID- 11884191 TI - An action research study exploring how education may enhance pain management in children. AB - The aim of this study was to explore and address the views of children's nurses in relation to their educational needs on pain management. Action research was the methodology used: focus groups were run to identify the problem of nurses' educational needs; action planning was used to develop a short programme of study for nurses to address identified needs. Evaluation was by questionnaire and semi structured interviews. Ten children's nurses attended the study day. All the nurses said they gained knowledge on the day--in particular assessment of pain and the individuality of the pain experience. The nurses felt that their new knowledge increased their confidence and contributed to them feeling assertive when managing children's pain. The study findings suggest that the current provision in relation to education programmes for children's nurses needs to be improved, in order to provide them with the knowledge and confidence to manage children's pain more effectively. PMID- 11884192 TI - Benefits and barriers for registered nurses undertaking post-graduate diplomas in paediatric nursing. AB - This paper presents one aspect of a larger study identifying key influences on curriculum redesign and development of a post-graduate diploma in advanced clinical nursing. The focus is on paediatric intensive care and general paediatric streams. Data presented here relate to registered nurses' perceptions of benefits and barriers when undertaking this post-graduate diploma. As well as interviews and focus group discussions with a number of nurses, data were collected through a self-administered questionnaire, A total of 885 surveys were distributed to nurses working in paediatric areas in five hospitals in Victoria, Australia. Of these, 391 were completed (response rate 44%). One hundred and thirty (33%) had post-registration or post-graduate paediatric qualifications. Perceived benefits of undertaking the post-graduate diploma mainly related to an increase in knowledge and experience and improvement of employment opportunities. Perceived barriers mainly related to financial and professional issues such as cost of the course, loss of salary, the lack of direct remuneration on completion of the course and a lack of promotional opportunities. It was of concern that several nurses expressed a belief that paediatric qualifications were unnecessary and that many believed their employers did not value the qualification. Several recommendations are suggested to address the main barriers. These include more flexibility in the provision of such courses and opportunities for financial assistance. PMID- 11884193 TI - The development of nursing students' spirituality and spiritual care-giving. AB - Nursing education programs are being increasingly challenged to incorporate spirituality and spiritual care-giving into the curriculum. The purposes of this study were to explore how students in a baccalaureate curriculum perceived their spirituality and spiritual health, and their perceptions of spiritual nursing care. Students in the first and fourth years of the program filled out a survey that included a spiritual well-being scale and several open-ended questions. Overall, students had a strong awareness of personal spirituality and a high level of spiritual health. They identified a number of behaviours and characteristics of the nurse that facilitated spiritual nursing care. Fourth year students demonstrated a more patient-centered approach to spiritual care. They placed less emphasis on the nurse's agenda and qualities and more on supporting the patient's beliefs. PMID- 11884194 TI - Exploring qualified nurses' perceptions of the relevance of education in preparation for their role in rehabilitation. AB - The increasing importance of rehabilitation in the health sector and the nurses' critical role therein make it essential that nurses have the right skills and knowledge to work effectively in rehabilitation settings. Drawing from a wider qualitative investigation of the role of the nurse within the multi-professional rehabilitation team, gaps in the skills and knowledge of qualified nurses working in rehabilitation settings are presented and ways to address them are proposed. Both pre- and post-registration education were found wanting. Only one third of nurses thought, in retrospect, that their pre-registration education had provided them with adequate skills and knowledge for their role in rehabilitation. A need for greater focus on rehabilitation per se and associated clinical skills was identified. Whilst post-registration education was highly valued, substantial difficulties accessing relevant courses were noted. In-service training and ad hoc learning 'from experience' and colleagues formed additional ways to develop hands-on skills. Benefits of better education included enhancing confidence, promoting inter-professional equality and improving client care. Potential ways to address some of these concerns included: adoption of a 'thread and module' approach and dedicated rehabilitation student placements, a nationally recognized multi-professional post-registration course, and an integration of work based learning with formal educational provision. PMID- 11884195 TI - Presentation skills workshops for nurses. AB - In the modern NHS, where there is increasing emphasis on evidence-based practice and clinical governance, all healthcare professionals need to be able to present the results of their professional activities. Traditionally, nurses have not been trained to present the results of their work either verbally or in writing. A series of presentation workshops for nurses were held in Scotland. The feedback and presentation activities of participants, in the year following the workshops, are reported here. Immediate verbal feedback indicated that the majority of those attending had enjoyed the workshops. Written feedback over the course of the following year showed that the learning needs of participants had largely been met and that many of those attending had gone on to present their own work. Presentations were both verbal and in writing at a variety of levels, ranging from local reports and presentations to journal articles and papers given at international conferences. Nurses responded well to the very practical, informal and non-threatening environment of the workshop. The development of presentation skills has wider implications for raising awareness of and sharing best practice in nursing care. PMID- 11884196 TI - Occupational health teaching for pre registration nursing students. AB - The amount of time spent teaching occupational health and the methods used to teach this subject in the pre registration nursing programmes in the UK was surveyed. Questionnaires were sent to the Deans of all 66 Schools of Nursing identified from the UCAS list in 2000. Forty six responded. Five returns were rejected because they did not meet the study criteria, giving a final response rate of 67%. The 41 schools were providing 33 Nursing Diploma and 31 Nursing Degree courses. The results indicate that occupational health is taught on the majority of Nursing Diploma (88%) and Degree (80%) courses. The main method of teaching is by lectures, with a smaller number of courses offering the opportunity for seminars and project work. However, the subjects covered under this heading frequently relate to the occupational health and safety issues that nurses need to know about in order to protect their own health and safety. It is rare that the broader concept of how a patient or client's health can be affected by their work or how their health may affect their ability to work, remain in employment or, for populations, its impact on sustainable development at the community level, are addressed. It is recommended that all nurses should receive some training in the broader concepts of occupational health at the pre registration level so that they can develop an awareness of the relationship between work and health, at both the individual and community level. This survey parallels a similar exercise carried out with medical students that identified a worrying decline in the teaching of occupational health at the undergraduate level. PMID- 11884197 TI - Student nurse satisfaction levels with their courses: part I -- effects of demographic variables. AB - Recent years have seen an increased emphasis on the monitoring of the quality of learning and teaching in higher education. This has led to more attention being paid to evaluating student perceptions of their learning experiences. This article (Part I of two on the subject) examines the factors that influence student satisfaction with their modular courses. Seven factors categorized into two sets are scrutinized: Part I investigates the effects of four demographic variables, while Part II explores the effects of three academic (educational) related variables. In this report, the study research design, tool and sample are described, and an analysis of the demographic variables of gender, disability, ethnicity and age-bracket is undertaken, whereby their effects on performance and satisfaction levels of students are considered. Employing a questionnaire, data from 460 students attending various multidisciplinary health care modules at the School of Health Care, Oxford Brookes University for the first term of the academic year 2000/2001 were analysed. The questionnaire was found to be reliable and there were no differences in satisfaction between students who provided their registration numbers and those who did not, suggesting that the results are generalizable to the greater student population of the School of Health Care. The study found no differences in performance or satisfaction levels according to gender, but disabled students indicated less satisfaction although the differences were not significant. Fewer students of 'non-white' ethnicity felt that their learning experience was intellectually stimulating, and that the information about their modules was satisfactory and readily available. Student age significantly predicted performance and 'mature' students performed better and showed higher satisfaction than 'traditional' students. The findings and their implications for nurse education and curriculum design are discussed, with support from a growing number of studies investigating student nurse satisfaction and quality issues of learning and teaching in higher education. PMID- 11884198 TI - Student nurse satisfaction levels with their courses: Part II--effects of academic variables. AB - The degree of student satisfaction with their educational experiences is an important dimension in the assessment of institutional effectiveness. All nursing education teams are currently working on ways to improve the quality of their educational provision and increase the satisfaction of their students. This is the second of a two-part article on the factors that influence student satisfaction with their courses. Part I examined how the four demographic features of gender, disability, ethnicity and age influenced student satisfaction and their performance on nursing modules. This paper complements Part I by examining the effects of three educational factors (academic level of the module, mode of study, and the qualification aim) on the accomplishment and satisfaction levels of 460 students attending multidisciplinary health care modules at the School of Health Care, Oxford Brookes University, 2000/2001. The study found that in contrast to Level 1 students, Level 3 participants felt the need for modules to stimulate more interest, that module teams be more skilled and knowledgeable and that library resources be more abundant. The findings also suggested that part-time students required significantly more attention than full-time participants in terms of the need for smaller seminar groups that would facilitate contributions and decrease inhibitions among students, and were more concerned with the utility and relevance of their learning in relation to their chosen professions. Diploma participants had the highest satisfaction followed by the BA and then the BSc students. The findings raise issues which are of interest to academic staff and nursing students, and the implications for nurse education and curriculum design are discussed within the context of student nurse satisfaction and quality issues of learning and teaching in higher education. PMID- 11884203 TI - Pharmacology of MEN 11467: a potent new selective and orally- effective peptidomimetic tachykinin NK(1) receptor antagonist. AB - We have investigated the pharmacological properties of MEN 11467, a novel partially retro-inverse peptidomimetic antagonist of tachykinin NK(1) receptors. MEN 11467 potently inhibits the binding of [(3)H] substance P (SP) to tachykinin NK(1) receptors in the IM9 limphoblastoid cell line (pK(i) = 9.4 +/- 0.1). MEN 11467 is highly specific for the human tachykinin NK(1) receptors, since it has negligible effects (pK(i) <6) on the binding of specific ligands to tachykinin NK(2) or NK(3) receptors and to a panel of 30 receptors ion channels unrelated to tachykinin receptors. The antagonism exerted by MEN 11467 at tachykinin NK(1) receptors is insurmountable in saturation binding experiments, both K(D) and B(max) of SP were significantly reduced by MEN 11467 (0.3-10 nM). In the guinea pig isolated ileum, MEN 11467 (0.03-1 nM) produced a nonparallel rightward shift of the concentration-response curve to SP methylester with a concomitant reduction of the Emax to the agonist (pK(B) = 10.7 +/- 0.1). Moreover the antagonist activity of MEN 11467 was hardly reversible despite prolonged washout. In vivo, MEN 11467 produced a long lasting (> 2-3h) dose-dependent antagonism of bronchoconstriction induced by the selective tachykinin NK(1) receptor agonist, [Sar(9), Met(O(2))(11)]SP in anaesthetized guinea-pigs (ID(50)s' = 29+/-5, 31+/ 12 and 670+/-270 microg/kg, after intravenous, intranasal and intraduodenal administration, respectively), without affecting bronchoconstriction induced by methacholine. After oral administration MEN 11467 produced a dose-dependent inhibition of plasma protein extravasation induced in guinea-pig bronchi by [Sar(9), Met(O(2))(11)] (ID(50) = 6.7 +/- 2 mg/kg) or by antigen challenge in sensitized animals (ID(50) = 1.3 mg/kg). After i.v. administration MEN 11467 weakly inhibited the GR 73632-induced foot tapping behaviour in gerbil (ED(50) = 2.96 +/- 2 mg/kg), indicating a poor ability to block central tachykinin NK(1) receptors. These results demonstrate that MEN 11467 is a potent, highly selective and orally effective insurmountable pseudopeptide antagonist of peripheral tachykinin NK(1) receptors with a long duration of action. PMID- 11884204 TI - Binding of chimeric NPY/galanin peptides M32 and M242 to cloned neuropeptide Y receptor subtypes Y1, Y2, Y4, and Y5. AB - Ligand binding to neuropeptide Y (NPY) receptors Y1, Y2, Y4, and Y5 from guinea pig was investigated using the two NPY-galanin hybrids M32 (galanin1-13-NPY25-36 amide) and M242 ([D-Trp(32)]M32). The affinity of M32 for Y1, Y2, and Y4 receptors was 13, 4, and 30nM, respectively, similar to that of NPY18-36 and NPY22-36 but 40-fold to 300-fold lower than the affinity of intact porcine NPY. M242 bound to the Y1, Y2, and Y4 receptors with 9-fold to 20-fold lower affinity than did M32. The affinities of M32 and M242 for Y5 were 400 and 800 nM, respectively. Thus, M32 seems to gain affinity relative to both of its constituent peptide portions although the NPY25-36 part may be sufficient for NPY receptor recognition, especially at the Y2 receptor. This suggests that the galanin portion of M32 influences and/or stabilizes the conformation of the NPY portion, similar to the effect seen for the NPY portion of M32 in binding to galanin receptors. PMID- 11884206 TI - Colocalization of vasopressin and oxytocin in hypothalamic magnocellular neurons in water-deprived rats. AB - The posterior lobe hormones vasopressin and oxytocin are expressed in mutually exclusive sets of magnocellular hypothamalic neurons. However, under certain functional conditions a partial coexpression has been observed. In the present study we subjected adult rats to long-term osmotic stress by water deprivation for up to 3 days. After 3 days, a marked reduction of vasopressin immunostaining was observed in the paraventricular and supraoptic nuclei as compared with controls. Coexistence of oxytocin and vasopressin occurred in a portion of the magnocellular neurons. Many fibers of the hypothalamic-neurohypophyseal tract contained both peptides. Rehydration for 24 h after 3 days of thirsting resulted in a light recovery of vasopressin immunoreactivity with almost none magnocellular neurons containing both nonapeptides. Our findings indicate that magnocellular hypothalamo neurohypophysial neurons are capable of oxytocin and vasopressin coexpression upon extended osmotic stress. PMID- 11884205 TI - NK(3) receptors in the feline nucleus tractus solitarius are not involved with the muscle pressor response. AB - Isometric muscle contractions cause an increase in mean arterial pressure and heart rate. Previously, we showed that substance P (SP) is released from sites in the feline medial nucleus tractus solitarius (mNTS) in response to isometric muscle contractions, and that it most likely interacted with NK(1) tachykinin receptors at these sites. This study was undertaken to determine whether other tachykinin receptors in this area of the brainstem are involved with the muscle pressor response. Receptor autoradiography, using [(125)I]Bolton-Hunter SP and [(125)I] [MePhe(7)] neurokinin B to label NK(1) and NK(3) receptors, respectively, indicated that NK(3) tachykinin receptors are as abundant as NK(1) and NK(3) receptors, respectively, indicated that NK(3) tachykinin receptors are as abundant as NK(1) receptors in this region of the feline brainstem Injections of the specific NK(3) receptor antagonist, SR 142801 (0.1 to 10 microM) into the mNTS did not modify the pressor response or the heart rate response to isometric muscle contractions. Injection of SR142801 into the NTS prior to the injection of the NK(1) antagonist, GR82334 did not affect the action of GR82334 to attenuate the muscle pressor reflex. We conclude that NK(3) receptors in the NTS are not involved with the regulation of cardiovascular function during activation of the muscle pressor response. PMID- 11884207 TI - Neuropeptide Y (NPY) mRNA in rat brain tissue: effects of decapitation and high energy microwave irradiation on post mortem stability. AB - mRNA recovery from brain tissue is affected by time-interval from death to inactivation of tissue, and may depend on sacrificial method. Sacrifice by high energy microwave irradiation increases recovery of intact neuropeptides and proteins, and it has been suggested that this may be valid also for neuropeptide mRNAs. We therefore compared post-mortem NPY mRNA recovery following decapitation or microwave irradiation. Total RNA yield was significantly higher in tissue from decapitated rats. A decline in NPY mRNA (amol/mg tissue) over time, presumably reflecting degradation, was found in frontal cortex, hippocampus and striatum. Following high-energy microwaves, NPY message levels were higher in occipital cortex, lower in the hypothalamus, and unaltered in the other brain regions examined. These results show that post-mortem processes contribute to estimates of NPY mRNA levels obtained using standard methods for obtaining brain tissue from experimental animals and raise the question whether different pools of NPY mRNA might be differentially affected by post-mortem degradation. A general protective effect of high-energy radiation against degradation is not supported. PMID- 11884208 TI - Circulating levels of neuropeptides (CGRP, VIP, NPY) in patients with fulminant hepatic failure. AB - The present study investigated the circulating levels and cerebral fluxes of calcitonin gene-related peptide (CGRP), vasoactive intestinal peptide (VIP), and neuropeptide Y (NPY) and their relation to cerebral blood flow (CBF) during normoventilation and hyperventilation in patients with fulminant hepatic failure (FHF). Sixteen patients with FHF were studied and compared to six patients with cirrhosis of the liver. CBF was measured by the (133)Xe wash-out technique. Blood samples were obtained simultaneously from the artery and internal jugular bulb. Concentrations of CGRP and VIP were higher in FHF than in cirrhosis, 87 (55-218) vs. 29 (21-42) pmol/L, and 11 (6-29) vs. 5 (3-9)pmol/L, respectively. NPY was normal, none of the measures were related to CBF, and there was no detectable net brain fluxes. Hyperventilation did not alter any of the measures. CGRP and VIP in FHF seem to reflect hemodynamic changes in the systemic rather than in the cerebral circulation. PMID- 11884209 TI - Salt-loading increases vasopressin and vasopressin 1b receptor mRNA in the hypothalamus and choroid plexus. AB - The choroid plexus plays a pivotal role in the production of cerebrospinal fluid (CSF). Messenger RNA (mRNA) transcripts encoding arginine vasopressin (AVP) and the vasopressin 1b receptor (V(1b)R) are found in various structures of the central nervous system, including the choroid plexus. The present study measured AVP and V(1b)R mRNA production in response to plasma hyperosmolality. Compared to rats maintained on water, 2% salt-drinking rats had increased levels of AVP and V(1b)R mRNAs in the supraoptic (SON) and paraventricular (PVN) nuclei of the hypothalamus and in the choroid plexus. The increase in V(1b)R mRNA in the SON and PVN as a result of plasma hyperosmolality may reflect changes in receptor production that, in turn, have a role in AVP autoregulation of hypothalamic magnocellular neurons. The increase of AVP and V(1b)R mRNAs in the choroid plexus further shows the involvement of AVP in the regulation of brain water content and cerebral edema. PMID- 11884210 TI - Calcitonin gene related peptide in familial dysautonomia. AB - Familial dysautonomia (FD) patients have diminished sensory C-fibers. Calcitonin gene related peptide (CGRP) is a widely distributed neuropeptide and prominent neurotransmitter in C-fibers. We show that plasma CGRP levels measured by radioimmunoassay is significantly lower in 51 FD patients compared to controls (P<0.001). In 11/51 FD patients with FD crisis and in 19/51 FD patients with pneumonia, the mean CGRP levels rose significantly as compared to their baseline (P<0.003, P<0.001, respectively). The deficiency of CGRP in FD patients is consistent with their depletion of C-fibers, and may explain some of their symptoms, either directly or via modulation of sympathetic activity. PMID- 11884211 TI - A novel concept to preserve the beneficial effects of hormone replacement therapy in bilaterally female ovariectomized rats: role of lovastatin therapy. AB - Estrogen replacement therapy (ERT) is claimed to reduce cardiovascular mortality by about 50% in postmenopausal women. This improvement is caused by favorable changes in lipid and lipoproteins metabolism, however, it also increases the incidence of the endometrial hyperplasia. Addition of progestin to ERT, referred to as hormone replacement therapy (HRT), has been shown to successively reduce this risk to the endometrium. Unfortunately, it has an adverse effect on high density lipoprotein cholesterol (HDLC) concentration, thus compromising the benefits of ERT. Therefore the issue here whether HRT given alone and/or concomitantly with 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (lovastatin) could exert any significant additional favorable effect on the lipid profile in bilaterally ovariectomized female rats. Sixty female Wistar rats were ovariectomized and treated with ERT (0.625 mg kg (-1)estradiol, E (2), IM every 2 weeks), HRT (estradiol plus progesterone, E (2)+ P, 0.625 mg kg (-1)estradiol and 5 mg progesterone kg (-1) respectively, IM every 2 weeks), and lovastatin (20 mg kg (-1)day (-1)orally) plus HRT (L + HRT) for 6 weeks. Blood aliquots were collected for serum and plasma separation. Serum vitamin E and plasma levels of C-reactive protein (CRP), nitric oxide (NO), lipid profile, and the susceptibility of non-HDLC to oxidation were determined. Moreover, thoracic aortas were dissected and directed for measurement of its lipid peroxide and NO contents. Treatment of ovariectomized rats with HRT showed a significant decrease ( P< 0.0001) in HDLC concentration compared to the group treated alone with ERT and increase ( P< 0.0001) in CRP levels compared to ovariectomized rats. HDLC and CRP are two powerful and significant predictors for increased cardiovascular risk in postmenopausal women. Addition of lovastatin as a complementary therapy to HRT revealed a significant 27% increment in HDLC and 48% decrement in CRP concentrations. Moreover, it significantly increased vitamin E, each of plasma and tissue content of NO and decreased atherogenic indexes (TC/HDLC, LDLC/HDLC), aortic lipid peroxide and susceptibility of non-HDLC to oxidation. In conclusion, this current study demonstrated that lovastatin together with continuous combined HRT seems to be more effective in the secondary prevention of coronary heart disease not only due to lipid lowering properties but also related to several other additive effects such as modification of endothelial function and inflammatory responses. PMID- 11884212 TI - l-carnosine and verapamil inhibit hypoxia-induced expression of hypoxia inducible factor (HIF-1 alpha) in H9c2 cardiomyoblasts. AB - Contractile failure of myocardial cells is a common cause of mortality in ischemic heart disease. In response to hypoxic conditions, cells upregulate the activity of hypoxia-inducible factor 1 (HIF-1) and express a number of genes encoding proteins that either enhance O (2)delivery or increase cellular ATP levels. HIF-1 is a heterodimer of bHLH-PAS proteins, HIF-1 alpha and HIF-1 beta. Both subunits are constitutively expressed under normoxic conditions, but HIF-1 alpha levels are kept low by proteolytic degradation, then stabilized under conditions of low O (2)by a mechanism that is poorly understood. Here we tested the hypothesis that expression of HIF-1 alpha in cardiac cells may be affected by two known cardioprotective agents. We tested l-carnosine, a naturally occurring dipeptide which has been shown to improve myocardial contractility during hypoxia, and verapamil, a calcium channel blocker frequently prescribed for the treatment of heart disease. The levels of HIF-1 alphamRNA remained relatively stable during time course hypoxia (1% O (2)) in H9c2 cardiomyoblasts, then increased slightly after 24 h. In cells pretreated with 1 microM carnosine, the levels of mRNA were transiently reduced, but then increased after 24 h similar to the controls. The levels of HIF-1 alpha protein increased rapidly in H9c2 cells within 30 min of hypoxia, but this induction was significantly reduced in cells treated with either carnosine or verapamil. In addition, treatment of cells with these agents further reduced the low levels of HIF-1 under normoxic conditions. These results suggest that l-carnosine and verapamil may affect the regulated proteolytic degradation of HIF-1 alpha in heart cells during hypoxia. PMID- 11884213 TI - Effects of glibenclamide, metformin and insulin on the incidence and latency of death by oubain-induced arrhythmias in mice. AB - This study was performed to investigate possible effects of glibenclamide, insulin and metformin on the death latency and incidence caused by a cardiac glycoside, oubain. Mice of both sexes were injected with oubain (i.p. 20 mg x kg (-1), glibenclamide (s.c. 0.1-10 mg x kg (-1), insulin (s.c. 0.3-3 U kg (-)) and metformin (i.p. 200 mg x kg (-1)) and combinations of the last three drugs with oubain. Death latency was measured and lethality incidence was calculated. Death was assessed by visual observation. Plasma glucose level was evaluated from the tail blood. Glibenclamide (0.1 mg x kg (-)) prolonged the latency from 11.3 plus minus 1.2 to 15.8 plus minus 1.8 min but failed to decrease the incidence of death. At higher doses (1--10 mg x kg (-1)) it had no effects on the latency or the incidence. 0.3 U kg (-1)insulin decreased the incidence from 73.7 to 33.3% ( P< 0.05) without affecting the latency. However the higher dose (3 U kg (-1)) did not have any effects on the incidence or the latency. Oubain increased blood glucose level from 114.1 plus minus 3.8 (control) to 152.1 plus minus 5.3 mg x dl (-1). Metformin (200 mg x kg (-1)) did not affect either the latency or the incidence of death. While metformin did not decrease plasma glucose, insulin and higher doses of glibenclamide (1--10 mg x kg (-1)) markedly lowered glucose in blood. However, at the dose of 0.1 mg x kg (-1)glibenclamide did not alter the glucose level in the blood but prevented oubain from increasing it. Insulin (0.3 U kg (-1)) and, to some extent, glibenclamide (0.1 mg x kg (-)) but not metformin could be effective antiarrhythmic agents against oubain-induced arrhythmias. PMID- 11884214 TI - Protective effects of poly (ADP-ribose) synthase inhibitors on digoxin-induced cardiotoxicity in guinea-pig isolated hearts. AB - Reactive oxygen species, generated and released during digoxin-induced cardiotoxicity, can produce an activation of poly (ADP-ribose) synthase (PARS). Our objective was to examine the effects of PARS inhibitors, 3-aminobenzamide (3 AB ) and nicotinamide, on digoxin-induced arrhythmias in guinea-pig isolated hearts. 3-AB (0.1-0.3 mM) and nicotinamide (0.3 mM) were added to the perfusion solution starting 10 min before digoxin infusion (8 microg x ml (-1)min ( 1)reaching the heart) and maintained throughout the experiments. Electrocardiograms and coronary perfusion pressure were recorded continuously, and digoxin-induced arrhythmias were determined. Nicotinamide markedly inhibited ventricular tachycardia (VT) incidence (from 100%, n= 7, to 29%, n= 7), and abolished ventricular fibrillation (VF) incidence. 3-AB (0.1 mM, n= 9) significantly decreased VT incidence from 100% ( n= 7) to 22% ( n= 9) and VF incidence from 86% ( n= 7) to 11% ( n= 9). Both nicotinamide and 3-AB (0.1 mM) markedly decreased number of ventricular ectopic beats (VEBs) and arrhythmia score. 3-AB at 0.3 mM ( n= 8) appeared to decrease the VT (to 63%) and VF incidence (to 38%), but these reductions did not reach statistically significance levels. Moreover, 3-AB at high concentration (0.3 mM) did not significantly modify the number of VEBs and arrhythmia score. There were no significant changes in coronary perfusion pressure, heart rate or pressure rate index measured at certain time points throughout the experiment in all groups. Our results suggest that PARS activation plays a role in the digitalis-induced cardiotoxicity in guinea-pig isolated hearts. PMID- 11884215 TI - Influence of hypertension on acetaldehyde-induced vasorelaxation in rat thoracic aorta. AB - Ethanol causes vasoconstriction and contributes to the development of hypertension. Acetaldehyde (ACA), the primary metabolite of ethanol, elevates blood pressure by releasing endogenous catecholamines. In vitro, ACA leads to vasorelaxation, although the response may vary among various vascular beds. This study examined the influence of hypertensive state on the ACA-induced vasorelaxant responsiveness. Ring segments of thoracic aorta were isolated from Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) and isometric tension development was measured. In aorta with or without intact endothelium, the contractile responses to KCl and norepinephrine were greatly attenuated, whereas vasoconstrictive response to 5-HT was enhanced, by hypertension. Vasorelaxant response to histamine was similar between WKY and SHR groups. ACA (1 30 mM) elicited endothelium-intact as well as -denuded vasorelaxation in a dose dependent manner in aorta from both WKY and SHR groups. Interestingly, the ACA induced endothelium-intact vasorelaxation was significantly diminished, whereas the ACA-induced endothelium-denuded vasorelaxation was significantly augmented, by hypertension. These data indicated that the ACA-induced vasorelaxant response, either endothelium-intact or-denuded, is altered by the hypertensive state. PMID- 11884217 TI - A novel evaluation of subcutaneous formulations by in vivo magnetic resonance imaging (MRI). AB - The applicability of magnetic resonance imaging (MRI) for non-invasive in vitro studies of parenteral vehicles without use of marker substances was investigated. A wide range of extended release (ER) formulations such as oils, a lipid emulsion, and water solutions of a cyclodextrin and block co-polymers were visualized in vitro and in vivo by a (1)H-MRI technique. The study included measurements in vitro in a beaker and by injections in pig flesh. In vivo studies were carried out in rats. The contrast of the vehicles vs the background material could be visualized and quantifications of vehicle dispersion and disappearance were performed on obtained in vivo data. A wide range of different vehicles suitable for s.c. ER delivery were tested, such as different oils, a lipid emulsion, and water solutions of a cyclodextrin and block co-polymers. The vehicle volume expansion in vivo was possible to follow. However, this was not generally applicable for all kinds of vehicle component. The tested co-polymers, Poloxamers, were one type of vehicle component that provides an excellent MRI signal. The in vitro tests predicted the suitability of this vehicle for in vivo MRI studies. In the in vivo study of the block co-polymer formulations the apparent vehicle volume increased to a peak value from an initial value close to the injected volume. Thereafter the volume diminished and no vehicle could be detected after 29 h after injection. MRI could be applied for measurements of the dispersion and disappearance of some vehicles at the site of injection after s.c. administration without use of contrast agents. PMID- 11884216 TI - Neurotoxicologic sequelae of tributyltin intoxication in rats. AB - Tributyltin oxide (TBTO) is a commonly used biocide. The purpose of this study is to correlate the toxicity of TBTO with the alterations of brain neurotransmitters and ATPases. TBTO was given by stomach tube to rats at either 37.5 or 75 mg x kg (-1)for 3 consecutive days. Nervous signs appeared in treated animals and the mortality reached 12 and 30%, respectively. The levels of brain dopamine, norepinephrine and serotonin decreased in a dose-dependent manner. The activities of brain total ATPase, Mg (2+)-ATPase and Na (+)/K (+)- ATPase were suppressed. The activity of Na (+)/K (+)- ATPase was more severely affected than that of Mg (2+)-ATPase. Histopathological changes in brain included hyperaemia, focal haemorrhages in vacuolated myelinated fibres, chromatolysis, or complete necrosis of neurons, degenerative changes, or complete absence of purkinje cells in the cerebellum. PMID- 11884218 TI - Inhibitory effect of herbal remedies on 12-O-tetradecanoylphorbol-13-acetate promoted Epstein-Barr virus early antigen activation. AB - For the past several years we have been evaluating natural products as potential cancer chemopreventive agents in a short term in vitro assay involving Epstein- Barr virus early antigen (EBV-EA) activation in Raji cells promoted by phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA). Because of the current interest in the use of herbal remedies, we considered examining them for their cancer chemopreventive activities, using their extracts with a view to uncovering such benefits (if any) these remedies might possess. Thirty-six extracts of 32 herbs belonging to 27 families in use as herbal remedies including those of gingko, black cohosh, echinacea, kava-kava, saw palmetto, turmeric, angelica, wild yam, cat's claw, passion flower, muira puama, feverfew, blueberry, chasteberry, licorice, nettle, golden seal, pygeum, ginger, valerian and hops were prepared and evaluated. Turmeric at a concentration of 10 microg x ml ( 1)exhibited the most potent anti-EBV-EA activity, which is ten times more than passionflower, that is next in the order of activity. At the concentration level of 100 microg ml (-1), several of the herbal remedies tested inhibited the EBV-EA in Raji cells exposed to the tumor promoter TPA (32 pM) by more than 90%. We also report for the first time the activities of 16 new medicinal plants as potential cancer chemopreventive agents. Since inhibitors of EBV-EA promoted by TPA in vitro have been shown to be effective anti-tumor promoting agents in laboratory animal models, our results indicate new and potential applications of these herbal remedies as cancer chemopreventive agents since they are already in clinical use in the human population. PMID- 11884219 TI - Differential effects of acidosis, high potassium concentrations, and metabolic inhibition on noradrenaline release and its presynaptic muscarinic regulation. AB - It was the aim of the present study to characterize the effect of single components of ischaemia, such as inhibition of aerobic and anaerobic energy production by combined anoxic and glucose-free perfusion (metabolic inhibition), high extracellular potassium concentrations (hyperkalaemia), and acidosis, on (1). the stimulated release of noradrenaline from the in situ perfused guinea-pig heart and (2). its presynaptic modulation by the muscarinic agonist carbachol. The release of endogenous noradrenaline from efferent cardiac sympathetic nerve endings was induced by electrical stimulation of the left stellate ganglion (1 min, 5 V, 12 Hz) and quantified in the coronary venous effluent by high performance liquid chromatography. Under control conditions, two consecutive electrical stimulations (S1, S2) elicited a similar noradrenaline overflow (S2/S1: 0.98 plus minus 0.05). After 10 min of global myocardial ischaemia overflow of endogenous noradrenaline was significantly reduced (S2/S1: 0.18 plus minus 0.03; P< 0.05). When studied separately, metabolic inhibition, hyperkalaemia (16 mM), and acidosis (pH 6.0) each markedly attenuated stimulated noradrenaline overflow (S2/S1: 0.65 plus minus 0.05, 0.43 plus minus 0.14, and 0.37 plus minus 0.09, respectively; P< 0.05). The muscarinic agonist carbachol (10 microM) inhibited stimulated noradrenaline release under normoxic conditions (S2/S1: 0.41 plus minus 0.07; P< 0.05). However, after 10 min of global myocardial ischaemia the inhibitory effect of carbachol on noradrenaline overflow was completely lost. Single components of ischaemia had a differential effect on presynaptic muscarinic modulation. Whereas hyperkalaemia (8-16 mM) did not affect muscarinic inhibition of noradrenaline release, carbachol lost its inhibitory effect during acidosis and metabolic inhibition. In conclusion, hyperkalaemia, metabolic inhibition, and severe acidosis each contribute to reduced overflow of noradrenaline after 10 min of myocardial ischaemia. However, presynaptic muscarinic inhibition of noradrenaline release was not affected by hyperkalaemia, but was sensitive to metabolic inhibition and low degrees of acidosis. PMID- 11884220 TI - Evidence for a role of nitric oxide in the mediation of antiproliferative UVA effects in keratinocytes. AB - Using cultured human keratinocytes, the present study investigates the role of nitric oxide (NO) in the mediation of the antiproliferative effects of ultraviolet light A (UVA). UVA treatment of cells (3-21 J cm (-2)) caused a time- and dose-dependent increase in nitrite formation in a micromolar range. This effect was accompanied by a decrease in DNA synthesis by 53.5%. Moreover, UVA treatment slightly reduced cell viability by 23.8%. Preincubation of keratinocytes with the NO scavenger 4,4,5,5-tetramethyl-imidazoline-1-oxyl-3 oxide (PTIO, 10-100 microM) or the NO synthase inhibitor N(G)-monomethyl-l arginine (l-NMMA, 30-300 microM) significantly diminished the UVA-induced increase in nitrite. PTIO as well as l-NMMA partially protected keratinocytes from UVA-induced antiproliferative effects and increased DNA synthesis by 67 or 49% of the control. The co-application of UVA irradiation (10 J cm (-2)) and the essential cofactor of NO synthases tetrahydrobiopetrin (BH4, 500 microM) led to an overadditive increase in the release of nitrite as well as to a decrease in DNA synthesis. These results imply that NO is involved in the antiproliferative UVA effects in keratinocytes. PMID- 11884221 TI - Acute effects of pentobarbital, thiopental and urethane on lung oedema induced by alpha-naphthythiourea (ANTU). AB - This study was designed to investigate the possible participation of urethane, pentobarbital sodium and thiopental sodium anaesthesia in the lung oedema induced by alpha-naphthylthiourea (ANTU), which is a well known noxious chemical agent in the lung. ANTU when injected intraperitoneally (i.p.) into rats (10 mg x kg (-1) i.p.) produced lung oedema as indicated by an increase in lung weight/body weight (LW/BW) ratio and pleural effusion (PE) reaching a maximum within 4 h. Administration of urethane prior to ANTU, at doses of 100 and 200mg(100g)(-1), elicited a significant and dose-dependent inhibition in LW/BW ratio and PE. Thiopental sodium at doses of 25, 50 mg x kg (-1), also produced a significant and dose-dependent inhibition of both parameters. Prior i.p. injection of pentobarbital sodium at a dose of 40 mg x kg (-1) elicited a significant inhibition in both parameters. These results suggest that i.p. urethane, thiopental sodium and pentobarbital sodium pretreatment have a prophylactic effect on ANTU-induced lung injury in rats. The possible role of the anaesthetics in lung oedema induced by ANTU and the possible underlying mechanisms are discussed. PMID- 11884222 TI - The effects of delta sleep-inducing peptide on incidence and severity in metaphit induced epilepsy in rats. AB - The effects of delta sleep-inducing peptide (DSIP) on metaphit- (1-(1(3 isothiocyanatophenyl)-cyclohexyl) (piperidine)-) induced audiogenic seizures in adult male Wistar rats were studied. The animals were divided into four experimental groups: 1. saline injected; 2. metaphit administered (10 mg x kg ( 1)); 3. metaphit administered plus DSIP injected (dose range 0.1-1 mg x kg (-1)) and 4. DSIP injected (1 mg x kg (-1)). Upon treatment, the rats were exposed to sound stimulation ( 100 +/- 3 dB, 60 s) at hourly intervals and the incidence and severity (running, clonus and tonus) of seizures were analyzed. In most animals, metaphit led to EEG abnormalities and elicited epileptiform activity recorded as spikes, polyspikes and spike-wave complex and increased power spectra. Time course studies revealed the peak of convulsive activity 7-12 h after the injection in metaphit-treated rats. DSIP acted as an anticonvulsant and the most potent anticonvulsive dose of 1 mg x kg (-1)significantly increased power spectra of deltawaves (2-11 h) in comparison with the saline-control group and decreased the incidence and duration of convulsive response, as well as mean seizure grade of metaphit-induced convulsions. These results suggest that DSIP should be considered as having potential anticonvulsant activity in this animal model. PMID- 11884223 TI - A study on the relationship between homocysteine and diabetic nephropathy in rats. AB - Hyperhomocysteinemia is known to be associated with many of the occlusive vascular diseases including ischemic heart disease. Elevated plasma total homocysteine (t-Hcy) is also remarkably common among patients with moderate to severe renal failure. The purpose of this study was to investigate the role of homocysteine (Hcy) in the pathogenesis of diabetic nephropathy in the rat. Additionally, any effect of aminoguanidine (AG), an inhibitor of advanced glycation end product (AGE) formation, on the onset of nephropathic symptoms and on the concentrations of Hcy was searched for. Diabetes was induced in male Wistar albino rats (6 months old) by a single injection of 50 mg x kg ( 1)streptozotocin (STZ) into the penile vein. Animals with blood glucose levels higher than 350 mg x dl (-1)72 h after STZ injection were included in the study. Age-matched rats receiving a single dose of citrate buffer served as controls. One half of the control and diabetic groups received AG via drinking water (1 g l (-1)). The experimental period lasted for ten weeks. Animals were killed by cardiac venipuncture after 24 hour urine samples were collected. Serum t-Hcy was quantified using HPLC, and urinary GAGs using the spectrophotometric 1,9-dimethyl methylene blue dye method. Serum glucose, protein, creatinine and total sulfydryl (t-SH) measurements, and urinary protein determinations were carried out spectrophotometrically. In diabetic rats, serum t-Hcy levels were significantly decreased (P< 0.001), and were negatively correlated with the urinary protein concentration (r= -0.67, P< 0.05). Urinary GAG levels were also increased in diabetic rats (P< 0.001). AG neither affected the t-Hcy levels, nor ameliorated the nephropathic symptoms. These results indicate that diabetic nephropathy is not linked to homocysteinemia in the rat. PMID- 11884224 TI - Diethyldithiocarbamate inhibition of galactosamine-induced hepatitis in rats. AB - Free-radical-mediated oxidant damage can contribute to acute hepatitis. Vitamin E, a classic antioxidant, has been tested as a therapy for rodent acute hepatitis, but the protection achieved has not been complete. This study demonstrated that in rats, sodium diethyldithiocarbamate (DDC), a potent antioxidant, strongly depressed galactosamine-induced hepatitis in terms of serum alanine amino transferase activities and bile acids, though not in terms of serum beta-glucuronidase activities. A potential limitation for DDC use in humans, inhibition of copper metalloenzyme activities, did occur at the DDC dose used here. However, these effects were not severe. Thus, DDC could make a useful short term therapeutic drug for acute hepatitis. PMID- 11884225 TI - Introduction: multifaceted roles of lipids and their catabolites in immune cell signaling. PMID- 11884226 TI - Phosphoinositide 3-kinase in immunological systems. AB - Phosphoinositide 3-kinases (PI3Ks) are an evolutionarily conserved family of signal transducing enzymes. A great variety of stimuli activate PI3K, leading to the transient accumulation of its lipid products in cell membranes. These lipids serve as second messengers to regulate the location and activity of an array of downstream effector molecules. In cells of the mammalian immune system, PI3K is activated by receptors for antigen, cytokines, costimulatory molecules, immunoglobulins and chemoattractants. Signaling via PI3K regulates immune cell proliferation, survival, differentiation, chemotaxis, phagocytosis, degranulation, and respiratory burst. Here we review our current understanding of PI3K signaling in leukocytes. PMID- 11884227 TI - Protein kinase B (Akt) regulation and function in T lymphocytes. AB - Protein kinase B (PKB) [1-5] is a serine/threonine kinase that is activated by cytokines, antigen receptors, the costimulator CD28 and chemokines in lymphocytes. [6-11] PKB is thus poised to contribute to a variety of immune activation responses. A number of functions have been ascribed to PKB in different cell lineages including the regulation of cell survival, cytokine gene induction and cell cycle progression. In the present article the mechanisms that control PKB activity in T lymphocytes will be reviewed and the function of this kinase in the immune system will be discussed. PMID- 11884228 TI - Signaling pathways of D3-phosphoinositide-binding kinases in T cells and their regulation by PTEN. AB - Phosphoinositide 3-kinases (PI3Ks) phosphorylate the D3 position of the myo inositol ring of inositol phospholipids, producing, amongst others, phosphatidylinositol-(3,4,5)-trisphosphate. This activity is opposed by the lipid phosphatase PTEN, which catalyzes the removal of this phosphate. Stimulation of PI3Ks is elicited by engagement of receptors for antigen, cytokines and chemokines, and by co-stimulatory molecules. Kinases and other enzymes containing pleckstrin homology domains are activated by binding to these phospholipids, affecting a variety of cellular processes that control lymphocyte function, including cell survival, proliferation, chemotaxis and cytoskeletal reorganization. This review highlights the signaling pathways of these kinases and other enzymes in T cells, their biological effects, and their regulation by PTEN. PMID- 11884229 TI - Regulation of the immune response by SHIP. AB - Multiple lines of experimental data indicate that SHIP1 is an important negative regulator of the immune system. SHIP1 has been demonstrated to control survival and proliferation, as well as differentiation. In the cases of some inhibitory receptors, such as Fc gamma RIIB1, the molecular mechanisms of control by SHIP1 are established. For other receptors, particularly activating receptors where SHIP1 appears to set activation thresholds, the mechanisms remain to be discovered. Further study on SHIP and other SHIP family members could be critical for our understanding the negative regulation in multiple hematopoietic lineages and the immune system as a whole. PMID- 11884230 TI - Phospholipase D and immune receptor signalling. AB - Immune receptors are coupled to the activation of phosphatidylcholine phospholipase D (PC-PLD) that hydrolyses phosphatidylcholine to generate phosphatidic acid and choline. As these receptors are also coupled to other signalling cascades, it has been difficult to define the precise cell activation events resulting from PLD activation in the absence of specific inhibitors. There is increasing evidence that phosphatidic acid acts as an intracellular signalling molecule regulating release of calcium from intracellular stores, sphingosine kinase and protein kinase C activation and membrane budding. Phosphatidic acid can also be rapidly converted into lysophosphatidic acid, diacylglycerol and arachidonates. PMID- 11884231 TI - Sphingolipids and the regulation of the immune response. AB - Over the last decade evidence has accumulated that sphingolipids are important and specific signalling molecules for cell-to-cell communication (mediator function) as well as for intracellular signalling processes (second messenger function). In addition, glycosylated sphingolipids are essential building blocks of rafts thereby participating in the initiation of receptor mediated signalling events. In immunology, processes such as T cell apoptosis, Th1 versus Th2 T cell differentiation, phagocytosis, and allergic excitability are either influenced or directly regulated by this class of lipids. Models such as the 'dual function concept' (differentiation of structural components versus signalling molecules) and the 'rheostat concept' (the balance of two or more sphingolipids is essential for the biological function) describe the multiple properties of these signalling molecules. PMID- 11884232 TI - Effects of currently used pesticides in assays for estrogenicity, androgenicity, and aromatase activity in vitro. AB - Twenty-four pesticides were tested for interactions with the estrogen receptor (ER) and the androgen receptor (AR) in transactivation assays. Estrogen-like effects on MCF-7 cell proliferation and effects on CYP19 aromatase activity in human placental microsomes were also investigated. Pesticides (endosulfan, methiocarb, methomyl, pirimicarb, propamocarb, deltamethrin, fenpropathrin, dimethoate, chlorpyriphos, dichlorvos, tolchlofos-methyl, vinclozolin, iprodion, fenarimol, prochloraz, fosetyl-aluminum, chlorothalonil, daminozid, paclobutrazol, chlormequat chlorid, and ethephon) were selected according to their frequent use in Danish greenhouses. In addition, the metabolite mercaptodimethur sulfoxide, the herbicide tribenuron-methyl, and the organochlorine dieldrin, were included. Several of the pesticides, dieldrin, endosulfan, methiocarb, and fenarimol, acted both as estrogen agonists and androgen antagonists. Prochloraz reacted as both an estrogen and an androgen antagonist. Furthermore, fenarimol and prochloraz were potent aromatase inhibitors while endosulfan was a weak inhibitor. Hence, these three pesticides possess at least three different ways to potentially disturb sex hormone actions. In addition, chlorpyrifos, deltamethrin, tolclofos-methyl, and tribenuron-methyl induced weak responses in one or both estrogenicity assays. Upon cotreatment with 17beta-estradiol, the response was potentiated by endosulfan in the proliferation assay and by pirimicarb, propamocarb, and daminozid in the ER transactivation assay. Vinclozolin reacted as a potent AR antagonist and dichlorvos as a very weak one. Methomyl, pirimicarb, propamocarb, and iprodion weakly stimulated aromatase activity. Although the potencies of the pesticides to react as hormone agonists or antagonists are low compared to the natural ligands, the integrated response in the organism might be amplified by the ability of the pesticides to act via several mechanism and the frequent simultaneous exposure to several pesticides. PMID- 11884233 TI - Cadmium- and mercury-induced intercellular adhesion molecule-1 expression in immortalized proximal tubule cells: evidence for a role of decreased transforming growth factor-beta1. AB - A definitive association between the aberrant expression of cytokines and adhesion molecules in renal failure has been established. In this regard a relationship between cytokine and adhesion molecule expression is suggested but has not been shown in models of proximal tubular cell injury. To investigate the impact of acute injury on the relationship between transforming growth factor beta 1 (TGF-beta1) and intercellular adhesion molecule-1 (ICAM-1) expression, two immortalized mouse proximal tubular epithelial cell lines were exposed to cadmium chloride or mercuric chloride (0-50 microM) for 0-8 h. ELISA and Western blot measured expression of secreted and intercellular TGF-beta1, respectively. Direct cellular ELISA or Western blot was used to assess ICAM-1 expression. Challenge with cadmium caused a greater loss of cell viability than did mercury. Interestingly, cadmium significantly decreased the amount of TGF-beta1 in the conditioned media. Although a similar trend was seen in mercury-challenged cells, no significant differences were observed. The decrease in TGF-beta1 in the culture media was not due to decreased expression of this cytokine, as intercellular levels were not affected by metal-induced injury. Significant increases in ICAM-1 protein expression were observed following cadmium and mercury challenge. The increase in ICAM-1 appears to be due to increased mRNA, as Northern blot analysis demonstrated increased message expression following a 4-h cadmium or mercury challenge. Supplementation of the culture media with exogenous TGF-beta1 decreased basal ICAM-1 expression and attenuated the cadmium-induced increase. These data suggest that metal-induced injury is associated with increased ICAM-1 expression. The mechanism of this induction may involve the decreased TGF-beta1 in the conditioned media following metal challenge. Taken together, these studies suggest a link between cytokine and adhesion molecule expression in renal injury. PMID- 11884234 TI - Decrease in K-ras p21 and increase in Raf1 and activated Erk 1 and 2 in murine lung tumors initiated by N-nitrosodimethylamine and promoted by 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - Recent evidence suggests that K-ras protooncogene protein p21 may have a tumor suppressive role in the context of development of lung adenocarcinoma. Levels of K-ras p21, raf-1, mitogen-activated protein kinases Erk 1 and 2, the phosphorylated-activated forms of Erk 1 and 2 (Erk 1P and 2P), and proliferating cell nuclear antigen (PCNA) were measured by immunoblotting in mouse lung tumors (5 to 9 mm in size) caused by N-nitrosodimethylamine (NDMA) and in control lungs. In tumors compared with normal lung, cell membrane-associated K-ras p21 was significantly decreased and cytosolic K-ras p21 increased. Total, membrane, and cytosolic raf-1 and Erk 1P and 2P were increased in tumors compared with normal lung. A single dose of 5 nmol/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) given after NDMA resulted in a significant 2.4-fold increase in tumor multiplicity. A significantly greater decrease in membrane-associated K-ras p21 and increase in total and membrane associated raf-1 occurred in the NDMA/TCDD tumors compared with the NDMA-only tumors. PCNA levels increased in tumors, a finding confirmed by immunohistochemistry, and correlated with tumor size after NDMA/TCDD treatment but not after NDMA only. The increase in raf-1 in the tumors was confirmed by immunohistochemistry, which also revealed an increase in raf-1-positive alveolar macrophages specifically associating with tumors from the earliest stages. These results suggest a possible tumor-suppressive function for K-ras p21 in lung and a positive role for raf-1 and Erk 1/2 in lung tumorigenesis. TCDD may promote tumors by contributing to downregulation of K-ras and stimulation of raf-1. PMID- 11884235 TI - Physiologically based modeling of the inhalation kinetics of styrene in humans using a bayesian population approach. AB - Animal studies have implicated styrene as toxic to the central nervous system and its major metabolite styrene-7,8-oxide as a carcinogen. Therefore, a reliable estimate of the metabolic capacity for styrene in humans is of interest. However, the available models describing styrene kinetics in humans lack rigorous statistical validation and also ignore the population variability in metabolism. The population variability may be estimated by the use of population models. Furthermore, the statistical validation of pharmacokinetic models may be improved by use of Bayesian methods. These two approaches may be combined and recently have been gaining interest in the toxicology literature. A population-based physiologically based pharmacokinetic (PBPK) model for styrene was developed. The model was calibrated to extensive human toxicokinetic data from three previous studies in which 24 volunteers were exposed to 50-386 ppm of styrene at rest and various levels of exercise. Model fitting was performed in a Bayesian framework using Markov chain Monte Carlo simulation. The uncertainty around the partition coefficients and metabolic parameters for styrene was reduced. The metabolic capacity for styrene in humans was estimated to be 0.92 micromol/l kg(-1), with a lognormal standard deviation of 1.66. The estimated Vmax is 40% higher than previously estimated, whereas the population standard deviation is estimated for the first time. PMID- 11884236 TI - Caffeine enhances the calcium-dependent cardiac mitochondrial permeability transition: relevance for caffeine toxicity. AB - Caffeine (1,3,7-trimethylxanthine), a compound present in beverages such as tea and coffee, is known to be toxic at high concentrations. Some of the observed clinical conditions include cardiovascular disease and reproductive disorders, among others. The possible toxic effects of caffeine on heart mitochondria are still poorly understood. The influence of caffeine on the mitochondrial permeability transition has not been clarified so far. The objective of this study was to investigate whether caffeine, at toxic concentrations, had any stimulating effect on the permeability transition of heart mitochondria isolated from Wistar rats, as well as whether it influenced mitochondrial respiratory parameters. Our results show that caffeine reduced mitochondrial ability to accumulate calcium by increasing the susceptibility of heart mitochondria to the opening of the transition pore. Caffeine not only hindered mitochondrial capacity to recover membrane potential after calcium addition but also increased the rate of calcium-dependent mitochondrial swelling and calcium-induced calcium release. The increased swelling was also observed in nonenergized mitochondria. Caffeine also showed a complex array of effects on heart mitochondrial bioenergetics, as evaluated by respiratory parameter measurements. We observed an increase in state 4 respiration and a depression in state 3 respiration, although no effect was observed on succinate-sustained mitochondrial membrane potential in the absence of calcium. Our work may be relevant to cardiovascular problems linked to caffeine toxicity and also to in vitro experiences involving caffeine-induced calcium release from the sarcoplasmic reticulum and uptake by mitochondria. PMID- 11884237 TI - Selective inhibitors of fatty acid amide hydrolase relative to neuropathy target esterase and acetylcholinesterase: toxicological implications. AB - Fatty acid amide hydrolase (FAAH) plays an important role in nerve function by regulating the action of endocannabinoids (e.g., anandamide) and hydrolyzing a sleep-inducing factor (oleamide). Several organophosphorus pesticides and related compounds are shown in this study to be more potent in vivo inhibitors of mouse brain FAAH than neuropathy target esterase (NTE), raising the question of the potential toxicological relevance of FAAH inhibition. These FAAH-selective compounds include tribufos and (R)-octylbenzodioxaphosphorin oxide with delayed neurotoxic effects in mice and hens plus several organophosphorus pesticides (e.g., fenthion) implicated as delayed neurotoxicants in humans. The search for a highly potent and selective inhibitor for FAAH relative to NTE for use as a toxicological probe culminated in the discovery that octylsulfonyl fluoride inhibits FAAH by 50% at 2 nM in vitro and 0.2 mg/kg in vivo and NTE is at least 100-fold less sensitive in each case. More generally, the studies revealed 12 selective in vitro inhibitors for FAAH (mostly octylsulfonyl and octylphosphonyl derivatives) and 9 for NTE (mostly benzodioxaphosphorin oxides and organophosphorus fluoridates). The overall in vivo findings with 16 compounds indicate the expected association of AChE inhibition with acute or cholinergic syndrome and >70% brain NTE inhibition with delayed neurotoxic action. Surprisingly, 75-99% brain FAAH inhibition does not lead to any overt neurotoxicity or change in behavior (other than potentiation of exogenous anandamide action). Thus, FAAH inhibition in mouse brain does not appear to be a primary target for organophosphorus pesticide-induced neurotoxic action (cholinergic or intermediate syndrome or delayed neurotoxicity). PMID- 11884238 TI - beta-Carotene reduces bleomycin-induced genetic damage in human lymphocytes. AB - We had previously shown in a human feeding study that ingestion of tomato and carrot juices decreases DNA breaks and oxidized pyrimidine bases in peripheral lymphocytes and enhances expression of glutathione S-transferase (GST) in a subpopulation of the volunteers. The aim of this study was to determine how the major carotenoids of these juices (beta-carotene or lycopene) could contribute to the observed antigenotoxicity. Physiological concentrations (2 microM) of water soluble beta-carotene and lycopene were incubated for 18-24 h with lymphocytes and then treated with bleomycin or H(2)O(2). Strand breaks, oxidized DNA bases, and persistence of damage (DNA repair) were measured by single-cell microgelelectrophoresis. GST-protein (GSTP1) was determined using an immunoassay and by measuring enzyme activity. HPLC analysis showed that beta-carotene was taken up by the cells after 24 h, and this was associated with a reduction of bleomycin-induced damage (29.11 +/- 1.86% tail intensity without versus 21.54 +/- 2.36% with beta-carotene). Lycopene was ineffective. The carotenoids did not modulate repair of bleomycin- and H(2)O(2)-induced damage and did not alter levels of oxidized pyrimidine bases nor GST expression. The results indicate that beta-carotene can enter the cell and protect against strand breaks but not against oxidized DNA bases. Therefore, beta-carotene accounts for only part of the protection observed in vivo with carotenoid-rich vegetable juices. PMID- 11884239 TI - Altered ethylbenzene-mediated hepatic CYP2E1 expression in growth hormone deficient dwarf rats. AB - Ethylbenzene (EB) effectively induces several hepatic P450 enzymes including CYP2E1 and CYP2B. Hypophysectomy diminishes the magnitude of EB-mediated induction of CYP2B. Although growth hormone (GH) plays a key role in sexual dimorphism of CYP2C11, its impact on EB-mediated P450 expression is still unknown. Because hypophysectomy leads to a depletion of multiple pituitary hormones besides GH, a study was designed to investigate the possible involvement of GH in EB-mediated hepatic P450 expression using GH-deficient dwarf rats as a more specific animal model. In these rats, pituitary GH was selectively reduced to about 10% of normal levels and other pituitary trophic hormones including thyroid-stimulating hormone, adrenocorticotropic hormone, luteinizing hormone, follicle-stimulating hormone, and prolactin are largely unchanged. Male control and HsdOla:DWARF-dw-4 (Harlan, UK) rats were subjected to a single ip injection of EB (10 mmol/kg). CYP2E1- and CYP2B-dependent activities, protein, and RNA levels were measured 10 and 24 h afterward. The results indicated that dwarf rats without EB exposure expressed higher CYP2E1. Although EB treatment induced CYP2E1 activity, protein, and mRNA both in controls and dwarf rats, the magnitude of the response to EB exposure was greater 10 h after the treatment in dwarf rats. Hypophysectomy also increased CYP2E1 protein induction by EB compared to intact rats. This effect was reversed by GH supplementation to hypophysectomized rats. Overall, responses of CYP2B to EB exposure in dwarf rats did not display basic differences from controls. In conclusion, the results demonstrate that (1) the suppression of CYP2B induction found in the multi-hormone-deficient HX rats is not found in the more specific GH-deficient rat model, confirming that GH does not have a major influence on CYP2B expression and (2) both hypophysectomized and GH-deficient rats show an altered inducibility of CYP2E1 after EB treatment. PMID- 11884240 TI - Enhancement of platelet aggregation and thrombus formation by arsenic in drinking water: a contributing factor to cardiovascular disease. AB - Arsenic in drinking water is a worldwide health problem that is associated with cardiovascular disease, but the cause is currently unknown. Arsenic effects on platelets, which are important in development of cardiovascular disease, were examined in vitro and in a drinking water study using a rat animal model. Trivalent inorganic arsenic (arsenite) induced in vitro aggregation when platelets were exposed to subthreshold challenge by thrombin and several other agonists in a concentration-dependent manner, with arsenite being the most potent form tested. Arsenite also induced significant increases in serotonin secretion, thromboxane A(2) formation, and adhesion protein expression in platelets. Consistent with the in vitro studies, 4-week ingestion of arsenite-contaminated drinking water resulted in enhanced arterial thrombosis. Human platelets showed similar responses, suggesting that the effects seen in animal experiments are applicable to humans. These results will provide new insights into the mechanism of arsenic-induced cardiovascular disease. They will also allow regulatory agencies to estimate risk from arsenic-induced cardiovascular disease and to determine if drinking water regulatory levels based on human cancer studies will protect against noncancer effects associated with cardiovascular disease. PMID- 11884241 TI - Direct comparison of the nature of mouse and human GST T1-1 and the implications on dichloromethane carcinogenicity. AB - Dichloromethane (DCM) is a hepatic and pulmonary carcinogen in mice exposed to high doses by inhalation. It has been shown previously that the incidence of liver and lung tumors does not increase in rats or hamsters exposed to the dihaloalkane under conditions similar to those that produced tumors in mice. The biological consequences of DCM exposure to humans is therefore uncertain. The carcinogenic effects of DCM in the mouse are caused by the interaction with DNA of a glutathione (GSH) conjugate that is produced by the class theta glutathione S-transferase T1-1 (GST T1-1). The species specificity is thought to be due to the greater amount of transferase activity in mouse target organs and specific nuclear localization of GST T1-1 in target cells. This paper directly compares the relative capacity and locality of DCM activation in mouse and human tissues. The results show that mouse GST T1-1 is more efficient in catalyzing the conjugation of DCM with GSH than the orthologous human enzyme. In addition, the mouse expresses higher levels of the transferase than humans in hepatic tissue. Histochemical analysis confirmed the presence of GST T1-1 in the nucleus of mouse liver cells. However, in human liver GST T1-1 was detected in bile duct epithelial cells and hepatocyte nuclei but was also present in the cytoplasm. Taking this information into account, it is unlikely that humans have a sufficiently high capacity to activate DCM for this compound to be considered to represent a carcinogenic risk. PMID- 11884243 TI - The pesticide methoxychlor disrupts the fusion of myoblasts into myotubes in skeletal muscle cell culture. AB - We studied the effect of the estrogenic pesticide methoxychlor (MXC) on skeletal muscle development using C2C12 muscle cell culture. Various concentrations of MXC or beta-estradiol (E) were added to the culture media. MXC (100 microM) disrupted myoblast fusion into myotubes, but 10 microM MXC or 10 microM E had no effect. Correlated with the diminished size of the myotubes, the clustering of acetylcholine receptors (AChRs) was inhibited by 100 microM MXC, but not by 10 microM MXC or 10 microM E. However, since clusters of AChR receptors did form, the postsynaptic clustering mechanism remained intact. Since E did not disrupt myoblast fusion into myotubes or the clustering of AChRs, we conclude that the abnormality induced by MXC is mediated by a mechanism of action that is independent of E. We believe this to be the first demonstration that MXC induces abnormal effects in the process of muscle development in skeletal muscle cell culture. PMID- 11884242 TI - Ultrafine airborne particles cause increases in protooncogene expression and proliferation in alveolar epithelial cells. AB - Exposure to ambient particulate matter (PM) is linked to increases in respiratory morbidity and exacerbation of cardiopulmonary diseases. However, the important components of PM and their mechanisms of action in lung disease are unclear. We demonstrate the development of dose-related proliferation and apoptosis after exposure of an alveolar epithelial cell line (C10) to PM or to ultrafine carbon black (ufCB), a component of PM. Ribonuclease protection assays demonstrated that increases in mRNA levels of the early response protooncogenes c-jun, junB, fra-1, and fra-2 accompanied cell proliferation at low concentrations of PM whereas apoptotic concentrations of PM caused transient increases in expression of fos and jun family members and dose responsive increases in mRNA levels of receptor interacting protein, Fas-associated death domain, and caspase-8. Significant increases in steady-state mRNA levels of protooncogenes and apoptosis-associated genes, TNFR-associated death domain, and Fas were also observed after exposure of epithelial cells to ufCB, but not fine carbon black or glass beads, respectively, suggesting that the ultrafine particulate component of PM is critical to its biological activity. PMID- 11884244 TI - Effects of gender on the cardiac toxicity elicited by chronic ethanol intake in rats. AB - Experiments were designed to determine if gender influences the cardiac toxicity elicited by chronic high-level ethanol intake. Male and female ethanol-preferring P-rats were allowed free access to drinking water or water containing 25% ethanol for 6 months. Left atrial preparations were then isolated, bathed in Krebs Henseleit solution (37 degrees C), and used to examine basal contractility at 3.0 Hz stimulation, the force-frequency relationship, and the positive inotropic response to the beta-adrenoceptor agonist isoproterenol. Basal contractile function was not affected significantly by ethanol in either gender; however, atria from ethanol-treated male rats displayed diminished contractility compared to control males when measured at slow stimulation frequencies (0.1 and 0.2 Hz), during post rest potentiation (at rest intervals of 20-60 s), and in response to higher concentrations of isoproterenol (> or =3 x 10(-7) M; EC50 values were not affected). In contrast, female atria showed no effect of chronic ethanol consumption. These data suggest that ethanol consumption diminishes the cardiac reserve in male, but not female rats. PMID- 11884245 TI - Differential inhibition of hepatic, pancreatic, and plasma fatty acid ethyl ester synthase by tri-o-tolylphosphate in rats. AB - Fatty acid conjugation of alcohols, catalyzed by fatty acid ethyl ester synthase (FAEES), results in the formation of lipophilic esters. Although the activity of FAEES is reported in almost all organs, including plasma, the interrelationship among various proteins expressing FAEES activity in different organs/tissues is not well understood. Earlier, we have reported an inhibition of FAEES activity in human hepatoma cells by tri-o-tolylphosphate (TOTP; serine esterase inhibitor). The present study was undertaken to further characterize the hepatic, plasma, and pancreatic FAEES in rats after ip injection of 10, 25, 50, or 100 mg/kg TOTP in corn oil or vehicle alone. After 18 h, animals were euthanized and FAEES activity in the plasma and postnuclear fractions of hepatic and pancreatic homogenates were assayed by measuring the ester formation following incubation with [1 (14)C]oleic acid and ethanol or methanol as substrates. Significant inhibition of FAEES activity was observed in hepatic postnuclear fraction. The esterase activity also showed a pattern similar to fatty acid ethyl and methyl ester synthesizing activity. A trend similar to hepatic synthesizing and hydrolyzing activities was also found in the plasma of TOTP-treated rats. However, no inhibition of synthetic activity toward formation of fatty acid ethyl or methyl esters or p-nitrophenyl acetate hydrolyzing activity was observed in the pancreas of rats after TOTP exposure. Our results also show that the protein expressing FAEES activity in the pancreas does not cross-react with antibodies to rat adipose tissue FAEES using Western blot analysis, which recognizes approximately 60- and approximately 84-kDa proteins in the liver and plasma, respectively. Furthermore, the inhibition in liver is at the functional level of enzyme as no change was observed between control and treated animals by immunohistochemistry. We conclude that fatty acid ethyl or methyl ester synthesizing enzyme(s) in the liver and plasma, which are inhibited by TOTP, are different from that present in the pancreas. PMID- 11884246 TI - Central effects of clozapine in regulating micturition in anesthetized rats. AB - BACKGROUND: We previously showed that systemic administration of the atypical neuroleptic clozapine in the rat altered a number of urodynamic variables and inhibited the external urethral sphincter. Since clozapine acts at several receptor types both at the periphery and the central nervous system, the site of action remained uncertain. Therefore, the purpose of this study was to determine the effects of central administration of clozapine on the bladder and the external urethral sphincter during cystometry and to examine differences in spinal versus supraspinal administration. We extended our observations by delivering clozapine centrally in anesthetized rats instrumented with either an intrathecal (L6-S1 spinal segment) or an intracerebroventricular (lateral ventricle) catheter. RESULTS: Clozapine decreased micturition volume and increased residual volume possibly by acting at a supraspinal site. Expulsion time and amplitude of the high frequency oscillations were reduced by clozapine possibly by acting at a spinal site. Bladder capacity was increased after central clozapine but probably due to a peripheral effect. Clozapine acting at spinal and supraspinal sites increased pressure threshold. Contraction time and peak pressure were not affected by clozapine. The EMG from the external urethral sphincter was also reduced following clozapine centrally and suggests a spinal and a supraspinal site of action. CONCLUSIONS: The results from the present study suggest that spinal and supraspinal central sites mediate clozapine's action in inhibiting expulsion parameters and the external urethral sphincter of the rat. Therefore, the reduction in the voiding efficiency observed after clozapine appears to be mediated by spinal and supraspinal sites. PMID- 11884247 TI - Can Australian general practitioners effectively screen for diabetic retinopathy? A pilot study. AB - BACKGROUND: Diabetes has been identified as one of the national health priority areas in Australia. After 20 years of diabetes most patients can be expected to develop diabetic retinopathy which, if undetected, is likely to cause significant visual loss or blindness. This paper reports on a pilot study aimed to test the ability of Australian GPs to clinically recognise diabetic retinopathy following a brief training intervention. METHOD: 17 GPs from a Brisbane Division of General Practice were recruited to participate in a clinical upskilling intervention pilot. Participant scores on clinical assessments were used to analyse GP sensitivity and specificity in screening for diabetic retinopathy. Results were compared with the NHMRC guidelines for acceptable screening accuracy. RESULTS: Ten of the 17 GPs (59%) achieved a screening sensitivity of 25% or less in the pre test, three (18%) a sensitivity of 50%, and four (23%) achieved a sensitivity of >or= 75%. In the post-test, all seventeen GPs achieved between 50 and 100% sensitivity. In the pre-test, thirteen (76%) GPs achieved a screening specificity of less than or equal to 50%, and four (23%) a specificity of 75 %. In the post test, four GPs (23%) rated a screening specificity of less than 50%, six (35%) achieved a specificity of 66%, and seven (41%) 100% specificity. CONCLUSION: 24% of GPs met the NHMRC diabetic retinopathy screening criterion prior to the workshop, and 94% following this brief training intervention. Australian GPs are capable of a much more significant role in community screening for diabetic retinopathy. PMID- 11884248 TI - Reporting of measures of accuracy in systematic reviews of diagnostic literature. AB - BACKGROUND: There are a variety of ways in which accuracy of clinical tests can be summarised in systematic reviews. Variation in reporting of summary measures has only been assessed in a small survey restricted to meta-analyses of screening studies found in a single database. Therefore, we performed this study to assess the measures of accuracy used for reporting results of primary studies as well as their meta-analysis in systematic reviews of test accuracy studies. METHODS: Relevant reviews on test accuracy were selected from the Database of Abstracts of Reviews of Effectiveness (1994-2000), which electronically searches seven bibliographic databases and manually searches key resources. The structured abstracts of these reviews were screened and information on accuracy measures was extracted from the full texts of 90 relevant reviews, 60 of which used meta analysis. RESULTS: Sensitivity or specificity was used for reporting the results of primary studies in 65/90 (72%) reviews, predictive values in 26/90 (28%), and likelihood ratios in 20/90 (22%). For meta-analysis, pooled sensitivity or specificity was used in 35/60 (58%) reviews, pooled predictive values in 11/60 (18%), pooled likelihood ratios in 13/60 (22%), and pooled diagnostic odds ratio in 5/60 (8%). Summary ROC was used in 44/60 (73%) of the meta-analyses. There were no significant differences in measures of test accuracy among reviews published earlier (1994-97) and those published later (1998-2000). CONCLUSIONS: There is considerable variation in ways of reporting and summarising results of test accuracy studies in systematic reviews. There is a need for consensus about the best ways of reporting results of test accuracy studies in reviews. PMID- 11884249 TI - Differences and similarities between patients with and without end-stage renal disease, with regard to location of intracardiac calcification. AB - It has been known for some time that mitral annulus calcification is common in end-stage renal disease (ESRD) patients on long-term dialysis, as well as in elderly patients without renal failure. However, a systematic comparison of cardiac calcification in these two types of patients has not yet been made. We examined two-dimensional echocardiograms in 33 patients with ESRD (mean age 66 +/ 10 years) and in 34 other patients with intracardiac calcification but no ESRD (mean age 69 +/- 9 years), with particular attention to precise anatomic location of calcification. Age was not significantly different in the two groups. The incidences of posterior mitral annulus calcification and aortic valve calcification were not significantly different in the ESRD and non-ESRD groups, though mitral annulus calcification tended to be larger in ESRD patients. Basal mitral leaflet calcification and papillary muscle calcification was much more common in the ESRD group. Calcification of intervalvar fibrosa and of tricuspid annulus were noted only in ESRD patients. PMID- 11884250 TI - Is left ventricular diastolic thickening documented during dobutamine and pacing stress echocardiography related to myocardial ischemia? An animal model study. AB - Transient increase in diastolic wall thickness (pseudohypertrophy) during pacing stress echocardiography has been reported in normal myocardium. To evaluate the occurrence of pseudohypertrophy and to investigate the contribution of myocardial ischemia on its production during pacing and dobutamine stress echocardiography, we produced a physiologically significant coronary stenosis in 14 open chest dogs. The stenosis in the circumflex artery was measured by quantitative coronary angiography (range: 50% to 89% reduction in luminal diameter), and no resting segmental wall-motion abnormalities were observed by epicardial echocardiography (short-axis, papillary level). In each study, dobutamine (5-40 microg/kg/min) and pacing (up to 260 beats/min) were performed randomly. Positivity of stress echocardiography tests was quantitatively determined by a significant (P < 0.05) reduction or failure to increase in absolute and percent systolic wall thickening in the myocardial area supplied by the stenotic artery as compared to the left anterior descending (LAD) artery-related areas. Diastolic wall thickness and left ventricular diastolic area were compared before and after each stress test in the circumflex and LAD artery-related regions. Pseudohypertrophy was observed in 57% and 86% of dogs for pacing and dobutamine, respectively, in the circumflex region, and in 50% and 64% in the LAD region. Despite its increased incidence in the circumflex region, the augmented diastolic wall thickness did not correlate with coronary stenosis severity or stress test positivity, but correlated inversely with changes in left ventricular diastolic area. In addition, it correlated directly with changes in heart rate only for pacing. In conclusion, pseudohypertrophy was a frequent finding during pacing and dobutamine stress echocardiography tests but was not related to myocardial ischemia in this animal model. PMID- 11884251 TI - Evaluation of left ventricular systolic and diastolic global function: peak positive and negative myocardial velocity gradients in M-mode Doppler tissue imaging. AB - OBJECTIVES: To evaluate a new indicator of left ventricular global function: Myocardial velocity gradient (MVG) M-mode Doppler tissue imaging (DTI). BACKGROUND: MVG is a new indicator of regional left ventricular function and global left ventricular diastolic function. However, it is unclear whether MVG also is an indicator of left ventricular global function in comparison with invasive indices. METHODS: We performed conventional imaging and M-mode DTI in 85 subjects and calculated MVG at the posterior wall. We obtained satisfactory images in 65 subjects, who we divided into three groups: Noninvasive study group, invasive study group, and hemodialysis group. The noninvasive study group was divided into three subgroups (a younger normal subgroup, an older normal subgroup, and a cardiomyopathy subgroup), and MVG was compared with indices of conventional imaging. In the invasive study group, we compared MVG and indices of conventional imaging with hemodynamic data (peak positive and negative dp/dt, and the time constant T) using a high fidelity micromanometer-tipped catheter. In the hemodialysis group, we compared indices before hemodialysis with those after hemodialysis. RESULTS: Peak positive MVG correlated well with peak positive dp/dt (r = 0.79), and this did not change with hemodialysis (P = 0.87). Peak negative MVG also correlated well with peak positive dp/dt and the time constant T (r = 0.88 and r = 0.80), and this did not change with hemodialysis (P = 0.97). CONCLUSIONS: Peak positive and negative MVG are sensitive and load-insensitive indicators of left ventricular function. PMID- 11884252 TI - Multicenter evaluation of SonoVue for improved endocardial border delineation. AB - OBJECTIVES: Two multicenter studies were conducted to evaluate the safety and efficacy of SonoVue as a contrast agent for enhanced left ventricular endocardial border delineation (LVEBD), and to compare the efficacy of SonoVue and Albunex in adult patients with a suboptimal, nonenhanced echocardiogram. BACKGROUND: The use of contrast to enhance echocardiographic assessment of LVEBD is well-established. SonoVue is a new microbubble contrast agent that contains sulfur hexafluoride. METHODS: Patients were randomized to receive four injections of SonoVue (0.5, 1, 2, and 4 ml), or two injections of Albunex and two injections of hand-agitated saline (0.08 and 0.22 ml/kg). Echocardiographic images were evaluated at the study centers and by four blinded, offsite reviewers for degree of left ventricle opacification (LVO), duration of contrast enhancement, and LVEBD. RESULTS: LVO scores were significantly higher for all doses of SonoVue. Patients with complete LVO ranged from 34%-87% for SonoVue and from 0%-16% for Albunex. The mean duration of useful contrast effect ranged from 0.8-4.1 minutes for SonoVue and < 15 seconds for Albunex. Mean increases in LVEBD scores ranged from 3.8-18.2 for SonoVue and 0.1-4.3 for Albunex. SonoVue (cumulative 7.5 ml dose) was well tolerated, with a safety profile similar to that observed in the control group. CONCLUSIONS: SonoVue is superior to Albunex for improving visualization of endocardial borders in patients with suboptimal noncontrast echocardiograms. Optimal increases in LVEBD, LVO, and duration of useful contrast effect were observed at the 2.0 ml dose of SonoVue. PMID- 11884253 TI - Left atrial appendage function and pulmonary venous flow in patients with nonrheumatic atrial fibrillation and their relation to spontaneous echo contrast. AB - This study analyzed the relation between frequency of left atrial appendage (LAA) contractions, pulmonary venous flow (PVF) parameters, and spontaneous echo contrast (SEC). Thirty-six patients (22 male, 14 female, mean age 61 plus minus 11 years) with nonrheumatic atrial fibrillation undergoing transesophageal echocardiography were studied. Doppler flow was obtained from both the LAA and the left upper pulmonary vein. Fourier analysis was applied to the LAA signal that exhibited the frequency of LAA contractions. LAA emptying velocity and PVF parameters were determined. There was no relation between velocity and frequency of LAA flow (r = 0.256, P = ns). Among LAA and PVF parameters, patients with left atrial SEC (n = 17) had a lower LAA velocity (16.8 +/- 10.8 cm/sec vs 35.6 +/- 13.2 cm/sec, P < 0.001), a larger LAA area (4.8 +/- 2.2 cm(2) vs 3.0 +/- 1.3 cm(2), P = 0.008), and a reduced systolic velocity time integral of PVF (3.4 +/- 2.2 cm vs 5.4 +/- 2.2 cm, P = 0.017) when compared with patients without SEC. Frequency of LAA contractions was similar between both groups (6.8 +/- 0.4 Hz vs 6.8 +/- 1.0 Hz, P = ns). In conclusion, the rate of LAA contraction does not correlate with LAA flow velocity and SEC. A low left atrial flow expressed by low LAA flow velocity and a reduction in systolic PVF is a major hemodynamic determinant for the occurrence of SEC. PMID- 11884254 TI - Quantitative assessment of aortic stenosis by three-dimensional anyplane and three-dimensional volume-rendered echocardiography. AB - Aortic stenosis is a challenge for three-dimensional (3-D) echocardiographic image resolution. This is the first study evaluating both 3-D anyplane and 3-D volume-rendered echocardiography in the quantification of aortic stenosis. In 31 patients, 3-D echocardiography was performed using a multiplane transesophageal probe. Within the acquired volume dataset, five parallel cross sections were generated through the aortic valve. Subsequently, volume-rendered images of the five cross sections were reconstructed. The smallest orifice areas of both series were compared with the results obtained by two-dimensional (2-D) transesophageal planimetry and those calculated by Doppler continuity equation. No significant differences were found between Doppler (0.76 +/- 0.18 cm(2)), 2-D echocardiography (0.78 +/- 0.24 cm(2)), and 3-D anyplane echocardiography (0.72 +/- 0.29 cm(2)). The orifice area measured smaller (0.54 =/- 0.31 cm(2), P < 0.001) by 3-D volume-rendered echocardiography. Bland-Altmann analysis indicated that for 3-D anyplane echocardiography, the mean difference from Doppler and 2-D echocardiography was - 0.04 +/- 0.24 cm(2) and - 0.06 +/- 0.23 cm(2), respectively. For 3-D volume-rendered echocardiography, the mean difference was 0.23 +/- 0.24 cm(2) and - 0.25 +/- 0.26 cm(2), respectively. In the subgroup with good resolution in the 3-D dataset, close limits of agreement were obtained between 3-D echocardiography and each of the reference methods, while the subgroup with poor resolution showed wide limits of agreement. In conclusion, planimetry of the stenotic aortic orifice by 3-D volume-rendered echocardiography is feasible but tends to underestimate the orifice area. Three-dimensional anyplane echocardiography shows better agreement with the reference methods. Accuracy is influenced strongly by the structural resolution of the stenotic orifice in the 3-D dataset. PMID- 11884255 TI - Echocardiographic indices of Doppler flow patterns compared with MRI or angiographic measurements to detect significant coarctation of the aorta. AB - Evaluation for the presence and severity of coarctation of the aorta (CoA) by two dimensional echocardiography alone can be difficult. The purpose of this study was to use Doppler velocity and pressure gradient half-time in systole and diastole to estimate CoA severity. Doppler echocardiograms of children with suspected CoA and either an aortic angiogram or thoracic magnetic resonance imaging (MRI) performed within 1 month of the echocardiogram were evaluated. Patients with patent ductus arteriosus, significant aortic insufficiency, long tubular CoA, or CoA outside the thorax were excluded. Measured Doppler variables, indexed for heart rate, included systolic velocity half-time (sVHTi), diastolic velocity half-time (dVHTi), systolic pressure half-time (sPHTi), and diastolic pressure half-time (dPHTi). For each of these variables, sensitivity and specificity to detect a significant CoA were determined. A significant CoA was defined as a ratio of the CoA diameter to the diaphragmatic aortic diameter of < 0.5 as imaged by MRI or angiography. Indexed systolic velocity and pressure half times were found not to be significant predictors for CoA. For the Doppler parameter dVHTi, using a critical value of > 200 msec indexed, we found a positive predictive value of 87% and a negative predictive value of 80%. The parameter dPHTi, using a critical value of > 75 msec indexed, demonstrated positive and negative predictive values of 92% and 79%, respectively. Measurement of dVHTi is a useful predictor for significant CoA, but the parameter dPHTi has an improved positive predictive value for detection of significant CoA. Systolic measurements of velocity or pressure half-times are not adequate to assess severity of CoA. PMID- 11884257 TI - Local thrombus as an isolated sign of traumatic aortic injury. AB - We report an unusual case of blunt traumatic aortic injury in which a mobile thrombus located at the isthmus was the only abnormality detected by transesophageal echocardiography. The case illustrates the high sensitivity of this diagnostic tool in aortic trauma and underscores the possibility of finding infrequent evidence of injury in subtle aortic lesions. PMID- 11884256 TI - Malignant thymoma invading the right atrium: a rare echocardiographic finding. AB - Malignant thymoma is a rare tumor. We report a case of malignant thymoma with intracaval extension and direct invasion of the right atrium presenting as superior vena cava syndrome. PMID- 11884258 TI - Acquired left ventricular-right atrial communication: Gerbode-type defect. AB - Left ventricular-right atrial (LV-RA) communications are rare intracardiac defects, often congenital in nature and clinically apparent during childhood. Acquired LV-RA shunts are encountered occasionally in the adult population as a result of a defect in the upper portion of the membranous ventricular septum. We describe the clinical and echocardiographic features of an elderly patient with an acquired LV-RA communication in the setting of an aortic composite valve graft and endocarditis. We also review the anatomical features and hemodynamic consequences of such defects. PMID- 11884259 TI - Atypical Bland-White-Garland syndrome: a 58-year-old woman with stenosis of the pulmonary origin of the left coronary artery. AB - The anomalous origin of the left coronary artery from the main pulmonary trunk (also known as Bland-White-Garland syndrome) is a rare congenital malformation that occurs in 0.4% of patients with cardiac anomalies. We present an adult case (a 58-year-old woman) of atypical Bland-White-Garland syndrome. The patient displayed a stenosis at the ostium of the anomalous origin of the left coronary artery and an aortopulmonary fistula. Using conventional angiography, it was not possible to differentiate between an anomalous origin of the pulmonary coronary artery and total stenosis of the left main coronary artery in combination with a pulmonary fistula. However, transesophageal echocardiography (TEE) succeeded in making this differential diagnosis. CONCLUSION: If there is subtotal or total occlusion, TEE can be used for detection of coronary vessel morphology, particularly in cases of coronary anomalies. PMID- 11884260 TI - Paradoxical embolism. PMID- 11884261 TI - Tissue Doppler-derived postsystolic motion in a patient with left bundle branch block: a sign of myocardial wall asynchrony. PMID- 11884262 TI - Transesophageal echocardiographic identification of thrombus producing obstruction of left pulmonary artery descending lobar branches and bronchial artery dilatation. AB - We report an elderly patient in whom a thrombus in the distal left pulmonary artery was shown by transesophageal echocardiography to extend and produce obstruction of the descending lobar branches as well as dilatation of the left bronchial artery. PMID- 11884264 TI - More on adrenal activity in the metabolic syndrome. PMID- 11884263 TI - Interaction between blood pressure, lipoproteins, angiotensin, and vascular disease. PMID- 11884265 TI - Vascular signaling pathways in the metabolic syndrome. AB - There are several potential cellular and molecular pathways whereby cardiovascular risk factors act through very specific signal transduction pathways in the formation of atherosclerosis, as seen often in the metabolic syndrome. Many examples point to multiple postreceptor defects in the insulin signaling pathway in vascular tissue, however, there are differences in the insulin receptor pathway in vascular tissue compared with skeletal muscle or fat. In addition to insulin receptors, insulin may affect atherosclerotic changes in the vascular cells via stimulation of insulin-like growth factor-1 receptors and their signaling pathway. Insulin also causes activation of the vascular renin angiotensin system in both vascular smooth muscle cells and endothelial cells. Insulin-activated tissue renin-angiotensin system leads to increased cell growth and contributes to the cause of atherosclerosis. The fact that agents that inhibit the renin-angiotensin system also block insulin-mediated renin angiotensin system expression and cell growth reinforces the potential implication of a vascular insulin-renin-angiotensin system pathway. Finally, novel substances such as the adipokines, factors produced from fat cells, reveal new risk factors in the metabolic syndrome and offer further evidence for a link between insulin resistance and accelerated atherosclerosis. PMID- 11884266 TI - Obesity-related hypertension: role of the sympathetic nervous system, insulin, and leptin. AB - Heightened sympathetic nervous system activity, hyperinsulinemia, insulin resistance, and hyperleptinemia contribute to obesity-related hypertension. However, the precise mechanism and sequence of events in this pathophysiologic event have not been clarified. This review concentrates on studies helping to clarify the mechanisms of blood pressure elevation associated with weight change, concentrating on the temporal changes in neuroendocrine factors that are known to control energy metabolism and blood pressure. A better understanding of the pathophysiologic mechanisms of obesity-related hypertension may help in prevention, treatment, and slowing of the cardiovascular complications of obesity. PMID- 11884267 TI - Cardiovascular and sympathetic effects of leptin. AB - Several studies have shown the association between obesity and hypertension. The pathophysiologic mechanisms of obesity-related hypertension remain unknown. Clinical and experimental studies have shown that obesity is associated with enhanced sympathetic nervous activity. Thus, sympathetic nerve activation seems to play a major role in obesity-associated hypertension. However, the factors responsible for this sympathoactivation have not been identified. Leptin is an adipocyte-derived hormone that promotes weight loss by reducing appetite and food intake and by increasing energy expenditure through sympathetic stimulation to brown adipose tissue. Leptin also produces sympathoactivation to kidneys, hindlimb, and adrenal glands, indicating that the obesity-associated increase in sympathetic nerve activity could be due in part to these sympathetic effects of leptin. However, obesity is associated with leptin resistance, since high circulating levels of leptin were observed in obese subjects. Recent evidences indicate that this leptin resistance could be selective with preservation of sympathetic effects despite the loss of metabolic action of leptin. This suggests divergent central pathways underlying metabolic and sympathetic effects of leptin. Several neuropeptides have emerged as potent candidate mediators of leptin action in the central nervous system, including the melanocortin system, neuropeptide Y, and cortico-trophin releasing factor. A detailed understanding of the multitude and complexity of integrated neuronal circuits and neuropeptide containing pathways in leptin action will help in understanding the pathogenesis of obesity and related disorders. PMID- 11884268 TI - The brain and salt-sensitive hypertension. AB - Genetically salt-sensitive rats, such as Dahl S and spontaneously hypertensive rats, show clear hypertensive responses to a high salt diet. Neural mechanisms play an essential role in salt-induced hypertension, and recent studies indicate that centrally induced sympathetic hyperactivity actually causes the hypertension. This review discusses the view that the renal genotype is not the only determinant of salt-induced sympathetic hyperactivity and hypertension, and that changes in genetic control of neuronal responses to cerebrospinal fluid Na(+) may play a primary role. PMID- 11884270 TI - Effects of gender on the renin-angiotensin system, blood pressure, and renal function. AB - Recent studies have identified key gender differences in cardiovascular function, renal hemodynamics, and the renin-angiotensin system. Extensive epidemiologic evidence has shown a clear gender difference in cardiovascular and renal disease progression, whereby female sex appears to be protective. The mechanisms underlying this phenomenon are unknown, but likely reflect the aforementioned sex differences in common pathophysiologic pathways. This review focuses on studies examining sex differences in these underlying pathways, which together may provide a plausible mechanism for the gender disparity in clinical outcomes. PMID- 11884269 TI - Renal afferents and hypertension. AB - The kidney and the autonomic nervous system are linked through renal nerves. Activation of efferent renal sympathetic nerves leads to changes in renal vascular resistance, renin release, and Na(+) and water retention. Evidence also exists indicating that the kidney is not just a target organ of sympathetic activity, but also acts as a sensor. Afferent renal nerves have been shown to carry information from renal chemoreceptors, which respond to changes in the composition of the interstitial fluid environment, and mechanoreceptors, which monitor hydrostatic pressure changes within the kidney, to the central nervous system. These afferent renal nerve inputs alter the activity of central integrative neuronal circuits that normally give rise to command signals that influence the function of effector organs. Renal receptors, through their connections at different levels of the neuraxis, are able to reflexly influence not only cardiovascular function through changes in sympathetic nerve discharge to a variety of vascular beds and the hypothalamic release of vasopressin, but also the function of the kidney. This increased sympathetic activity and hormonal release induced by activation of afferent renal nerves has been implicated in hypertension of diverse etiologies. PMID- 11884271 TI - Mechanisms of pressure natriuresis. AB - A central component of the feedback system for long-term control of arterial pressure is the pressure-natriuresis mechanism, whereby increases in renal perfusion pressure lead to decreases in sodium reabsorption and increases in sodium excretion. The specific intrarenal mechanism for the decrease in tubular reabsorption in response to increases in renal perfusion pressure appears to be related to increases in hemodynamic factors such as medullary blood flow and renal interstitial hydrostatic pressure (RIHP), and renal autocoids such as nitric oxide, prostaglandins, kinins, and angiotensin II. Increases in renal perfusion pressure are associated with significant increases in RIHP, nitric oxide, prostaglandin E2, and kinins, and decreases in angiotensin II. The mechanism whereby RIHP increases in the absence of discernible changes in whole kidney renal blood flow and peritubular capillary hydrostatic and/or oncotic pressures may be related to increases in renal medullary flow as a result of nitric oxide-induced reductions in renal medullary vascular resistance. Several lines of investigation support an important quantitative role for RIHP in mediating pressure natriuresis. Preventing RIHP from increasing in response to increases in renal perfusion pressure markedly attenuates pressure natriuresis. Furthermore, direct increases in RIHP, comparable to increases measured in response to increases in renal perfusion pressure, have been shown to significantly decrease tubular reabsorption of sodium in the proximal tubule and increase sodium excretion. The exact mechanism whereby RIHP influences tubular reabsorption is unknown, but may be related to alterations in tight junctional permeability to sodium in proximal tubules, redistribution of apical sodium transporters, and/or release of renal autacoids such as prostaglandin E2. PMID- 11884272 TI - Reactive oxygen species: roles in blood pressure and kidney function. AB - Oxidative stress in blood vessels and the kidney in hypertension can be induced by diverse vasoconstrictor mechanisms, including blockade of nitric oxide synthase and activation of angiotensin II type I receptors and thromboxane receptors. It can cause vasoconstriction via bioinactivation of nitric oxide, and by nitric oxide synthase independent mechanisms that include increased generation of endothelin-1 and the effects of superoxide anion and hydrogen peroxide on vascular smooth muscle cells. Oxidative stress can accompany hypertension in many models including the spontaneously hypertensive rat, the angiotensin II-infused rat, renovascular hypertension, the deoxycorticosterone acetate-salt model, and obesity-related hypertension. In the kidney, NADPH oxidase-generating superoxide anion is expressed in the vasculature, interstitium, juxtaglomerular apparatus, and the distal nephron. Much progress has been made in defining the pathways that intervene between agonist stimulation of blood vessels and reactive oxygen species-mediated contractile and renal functional responses in animal models in hypertension. PMID- 11884274 TI - Coronary calcium, race, and genes. PMID- 11884275 TI - Leukocyte recruitment into developing atherosclerotic lesions: the complex interaction between multiple molecules keeps getting more complex. PMID- 11884273 TI - Angiotensin II-mediated signal transduction pathways. AB - The renin-angiotensin system is one of the major cardiovascular systems that controls blood volume, peripheral vascular tone, and blood pressure. Recent studies indicate important roles for angiotensin II in inflammation, atherosclerosis, and congestive heart failure as well. It is gradually becoming clear that angiotensin II exerts effects on the cardiovascular system through several unique mechanisms, including the availability of two different angiotensin II receptors, recruitment of protein tyrosine kinase activity, and receptor tyrosine kinase transactivation. This review discusses the diverse mechanisms of angiotensin II-mediated signal transduction pathways and the various effects of angiotensin II on the cardiovascular system. PMID- 11884276 TI - Purinergic signaling and vascular cell proliferation and death. AB - Evidence for the role of purinergic signaling (via P1 and P2Y receptors) in the proliferation of vascular smooth muscle and endothelial cells is reviewed. The involvement of the mitogen-activated protein kinase second-messenger cascade in this action is clearly implicated, although details of the precise intracellular pathways involved still remain to be determined. Synergistic actions of purines and pyrimidines with growth factors occur in promoting cell proliferation. Interaction between purinergic signaling for vascular cell proliferation and cell death mediated by P2X7 receptors is discussed. There is evidence of the release of ATP from endothelial cells, platelets, and sympathetic nerves as well as from damaged cells in atherosclerosis, hypertension, restenosis, and ischemia; furthermore, there is evidence that vascular smooth muscle and endothelial cells proliferate in these pathological conditions. Thus, the involvement of ATP and its breakdown product, adenosine, is implicated; it is hoped that with the development of selective P1 (A2) and P2Y receptor agonists and antagonists, new therapeutic strategies will be explored. PMID- 11884277 TI - Matrix metalloproteinase inhibition impairs adipose tissue development in mice. AB - The effect of galardin, a broad-spectrum matrix metalloproteinase (MMP) inhibitor, was studied in mice kept on a high fat diet (HFD). Five-week-old male wild-type mice were fed the HFD (42% fat) for up to 12 weeks and were daily injected intraperitoneally with the inhibitor (100 mg/kg) or with vehicle. After 12 weeks of the HFD, the body weights of both groups were comparable, but the weight of the isolated subcutaneous (SC) or gonadal (GON) fat deposits was significantly lower in the inhibitor-treated group than in the control group (88 +/- 11 versus 251 +/- 66 mg, respectively, for SC fat [P<0.05]; 90 +/- 24 versus 217 +/- 30 mg, respectively, for GON fat [P<0.02]). The number of adipocytes was somewhat higher and the diameter was somewhat smaller (but not significantly) in adipose tissues of the inhibitor-treated group. Adipose tissue of the inhibitor treated mice contained more collagen than did that of the vehicle-treated mice (Sirius red-stained area of 42 +/- 2.6% versus 22 +/- 4.4%, respectively, for SC fat [P<0.05]; 21 +/- 5.1% versus 4.7 +/- 0.92%, respectively, for GON fat [P<0.01]); a distinct collagen-rich cap was formed around the inhibitor-treated tissue. In situ zymography with casein- or gelatin-containing gels confirmed a reduced MMP activity in SC and GON adipose tissues of inhibitor-treated mice. Thus, in this model, growth and development of adipose tissue appears to be limited by the formation of a collagen-rich matrix cap around the inhibitor treated tissue. These data suggest a functional role for MMPs in the development of adipose tissue. PMID- 11884278 TI - Insulin inhibits apoptosis of macrophage cell line, THP-1 cells, via phosphatidylinositol-3-kinase-dependent pathway. AB - Hyperinsulinemia has recently been reported as a risk factor for atherosclerotic diseases such as coronary heart disease; however, its precise mechanism is not well understood. To elucidate the role of insulin in the development of atherogenesis, we have investigated the effect of insulin on cell survival in macrophages, which are known to be important in the atherosclerotic process. Apoptosis was induced in macrophage cell lines derived from human monocytes or murine macrophages by serum starvation. Insulin administration retarded macrophage apoptosis by means of DNA laddering, dimethylthiazol diphenyltetrazolium bromide assay, and annexin V binding assay. Insulin also enhanced mRNA expression and protein production of the antiapoptotic Bcl-XL gene in a dose-dependent manner within the range of physiological concentrations. In the exploration of the signaling pathway involved in these antiapoptotic effects of insulin, pretreatment of cells with a specific inhibitor of phosphatidylinositol-3-kinase significantly suppressed insulin-mediated cell survival and insulin-induced Bcl-XL expression in macrophages. These data indicate that the survival effect of insulin on the apoptosis of macrophages is associated with the upregulation of Bcl-XL expression, and it may be mediated through the phosphatidylinositol-3-kinase signaling pathway. These mechanisms could be involved in the possible role of insulin in the development of atherosclerosis. PMID- 11884279 TI - Human coronary smooth muscle cells internalize versican-modified LDL through LDL receptor-related protein and LDL receptors. AB - Versican-like proteoglycans are the main component of the intimal extracellular matrix interacting with low density lipoprotein (LDL). The aim of this study has been to investigate the receptors involved in versican-modified LDL uptake by human vascular smooth muscle cells (VSMCs). We have found that versican-LDL interaction leads to the following: (1) monomeric LDL particles that are similar in size and electrophoretic mobility to native LDL but that have a higher capacity to induce intracellular cholesteryl ester (CE) accumulation and (2) fused LDL particles similar in size to those obtained by vortexing. The precipitable fraction of versican-LDL, composed of 50% monomeric and 50% fused LDL particles, induced a dose-response increase in the CE content of VSMCs. Anti LDL receptor antibody decreased the CE accumulation derived from monomeric LDL particles by 88 +/- 3% and that derived from the total precipitable fraction by 45 +/- 3%. Inhibition of LDL receptor-related protein expression by antisense oligodeoxynucleotides reduced the CE accumulation derived from the precipitable fraction by 65 +/- 2.8%, whereas it did not produce any effect on the CE accumulation derived from monomeric LDL. These results suggest that versican-LDL induces CE accumulation in human VSMCs by the LDL receptor (monomeric particles) and LDL receptor-related protein (fused LDL). PMID- 11884280 TI - Activation of big mitogen-activated protein kinase-1 regulates smooth muscle cell replication. AB - This study examined the activation of big mitogen-activated protein (MAP) kinase 1 (BMK1) in rat carotid smooth muscle cells (SMCs). Platelet-derived growth factor, fibroblast growth factor-2, sorbitol, and serum all increased the activation of BMK1 in rat carotid SMCs, whereas angiotensin II, phorbol esters, and tumor necrosis factor-alpha had only slight effects. With the exception of tumor necrosis factor-alpha, all these factors phosphorylated extracellular signal-regulated kinase (ERK)1/2. The MAPK kinase inhibitor (MEKI), U0126 (1 micromol/L), blocked ERK1/2 phosphorylation and at higher doses (5 micromol/L) blocked BMK1 phosphorylation. This inhibitor also blocked SMC DNA synthesis in a dose-dependent manner. When SMCs were transfected with an adenoviral construct expressing dominant mutant BMK1 and stimulated with fibroblast growth factor-2, a significantly smaller increase in cyclin D1 and cyclin A expression and in retinoblastoma factor phosphorylation was detected compared with the increase in cells transfected with an adenoviral construct expressing green fluorescent protein (GFP). SMC DNA synthesis was significantly blocked in the cells transfected with the dominant mutant BMK1. These data support the suggestion that BMK1 is important and necessary for mitogen-induced SMC proliferation. PMID- 11884281 TI - Increased plasmin and serine proteinase activity during flow-induced intimal atrophy in baboon PTFE grafts. AB - High blood flow causes intimal atrophy and loss of extracellular matrix in PTFE aortoiliac grafts. We have investigated whether matrix-degrading proteinases are altered in this baboon model of atrophy using zymography, western analysis, and a versican degradation assay. After four days of high flow, urokinase was increased and plasminogen activator inhibitor-1 was decreased in the intima. Plasminogen was increased after seven days. Pro-matrix metalloproteinase (MMP)-2, activated MMP-2, and proMMP-9 levels were modestly increased by high flow at 7 days, whereas MMP-3 and tissue inhibitor of metalloproteinases-1 were not altered. Extracts of 4-day high-flow intimas degraded more 35S-methionine-labeled versican than low-flow intimal extracts, and this activity was inhibited by AEBSF, a serine proteinase inhibitor, and a plasmin antibody. In contrast, this activity was not inhibited by the MMP inhibitor, BB-94 (Batimastat). These data suggest that serine proteinases, including plasmin, may be largely responsible for extracellular matrix degradation in this primate model of flow-induced intimal atrophy. PMID- 11884282 TI - Different effects of high and low shear stress on platelet-derived growth factor isoform release by endothelial cells: consequences for smooth muscle cell migration. AB - In the present study, we analyzed the effect of conditioned media (CM) from bovine aortic endothelial cells exposed to laminar shear stress (SS) of 5 dyne/cm2 (SS5) or 15 dyne/cm2 (SS15) for 16 hours on smooth muscle cell (SMC) migration. In response to CM from bovine aortic endothelial cells exposed to SS5 (CMSS5) and SS15 (CMSS15), migration was 45 +/- 5.5 and 30 +/- 1.5 cells per field, respectively (P<0.05). Similar results were obtained with SS of 2 versus 20 dyne/cm2 and also when SS of 5 and 15 dyne/cm2 lasted 24 hours. Platelet derived growth factor (PDGF)-AA levels in CMSS5 and CMSS15 were 9 +/- 7 and 18 +/ 5 ng/10(6) cells for 16 hours, respectively (P<0.05); PDGF-BB levels in CMSS5 and CMSS15 were 38 +/- 10 and 53 +/- 10 ng/10(6) cells for 16 hours, respectively (P<0.05). PDGF receptor alpha (PDGFRalpha) and PDGF receptor beta (PDGFRbeta) in SMCs were phosphorylated by CMSS15>CMSS5. In response to CMSS15, a neutralizing antibody against PDGF-AA enhanced SMC migration to a level comparable to that of CMSS5; in contrast, antibodies against PDGF-BB abolished SMC migration. Transfection of SMCs with a dominant-negative PDGFRalpha or PDGFRbeta increased or inhibited, respectively, SMC migration in response to CMSS15. Overexpression of wild-type PDGFRalpha inhibited SMC migration in response to CMSS5, CMSS15, or recombinant PDGF-BB (P<0.001). These results suggest that the ability of high SS to inhibit arterial wall thickening in vivo may be related to enhanced activation of PDGFRalpha in SMCs by PDGF isoforms secreted by the endothelium. PMID- 11884283 TI - Novel 5' exon of scavenger receptor CD36 is expressed in cultured human vascular smooth muscle cells and atherosclerotic plaques. AB - CD36, a member of the scavenger receptor family, is centrally involved in the uptake of oxidized low density lipoproteins (oxLDLs) from the bloodstream. During the atherosclerotic process, the lipid cargo of oxLDL accumulates in macrophages and smooth muscle cells (SMCs), inducing their pathological conversion to foam cells. Increased expression of CD36 occurs in human atherosclerotic lesions, and CD36 knockout mice show reduced uptake of modified LDLs and reduced atherosclerosis. Here, we describe a novel exon 1b and extended CD36 promoter in human SMCs. Exon 1b is specifically transcribed in activated aortic SMCs and mainly expressed in atherosclerotic plaques. Thus, switching to exon 1b transcription may be an important step for the activation of SMCs and their conversion to foam cells. Using an antisense oligonucleotide to exon 1b, we inhibit CD36 translation and highly reduce oxLDL uptake. The antisense to exon 1b does not affect CD36 in cell lines not expressing the new exon. The possibility of a novel antiatherosclerotic therapy and the use of exon 1b as a marker of atherosclerosis are discussed. PMID- 11884284 TI - Autosomal genome-wide scan for coronary artery calcification loci in sibships at high risk for hypertension. AB - Coronary artery disease (CAD) is the leading cause of mortality in the developed world. Although several CAD risk factors, including measures of lipid metabolism, obesity, and blood pressure, have a genetic basis, many genes for CAD susceptibility have yet to be identified. Coronary atherosclerosis is the major cause of CAD, but many with coronary atherosclerosis lack symptoms. Thus, a major limitation of using symptomatic CAD endpoints (eg, sudden coronary death, myocardial infarction) as a study outcome is substantial disease misclassification. Coronary artery calcification (CAC) is part of the atherosclerotic process and is an independent predictor of CAD endpoints. In the present study, CAC was noninvasively quantified by electron beam computed tomography. We performed genome-wide multipoint mode-of-inheritance-free linkage analysis on affected sib pairs, defined as being > or = the 70th sex- and age specific percentile for CAC quantity, in a sample of 29 families enriched for hypertension. Almost 95% of participants were asymptomatic for CAD. Our LOD score (log10 odds in favor of linkage) results provide evidence that chromosomal regions 6p21.3 (maximum LOD score=2.22, P=0.00070) and 10q21.3 (maximum LOD score=3.24, P=0.000057) may harbor genes associated with subclinical coronary atherosclerosis. PMID- 11884285 TI - Racial differences in coronary artery calcification in older adults. AB - Reports on race-related differences in coronary artery calcium (CAC) are just beginning to emerge and have not been well studied in the elderly. This study was undertaken to assess whether such differences exist and the relationship between CAC and cardiovascular risk factors in a cohort of elderly community-dwelling adults. CAC was measured by using electron-beam tomography in 614 adults (aged 67 to 99 years), of whom 59% were women and 23% were black. The median CAC score was lower in blacks than in whites for men (159 versus 787, respectively; P<0.001) and for women (134 versus 233, respectively; P=0.02) after adjustment for age, cardiovascular disease, and risk factors for cardiovascular disease, although this difference was stronger and remained significant among men only. Lower CAC scores were also observed in the subgroup of blacks with a history of myocardial infarction. The lower CAC scores in blacks compared with whites observed in this study is consistent with either a lower prevalence of coronary artery disease or a lower extent of calcification of coronary artery disease. PMID- 11884286 TI - Autoimmunity to human heat shock protein 60, Chlamydia pneumoniae infection, and inflammation in predicting coronary risk. AB - Heat shock protein 60 (Hsp60) and Chlamydia pneumoniae infection have both been associated with cardiovascular diseases. Our aim was to study the role of Hsp60 antibodies as coronary risk predictors and their association with C pneumoniae infection and inflammation. This was a prospective, nested, case-control study. The cases consisted of 239 middle-aged Finnish men who developed myocardial infarction or coronary death during the follow-up. Baseline levels of IgA and IgG antibodies to human-specific and C pneumoniae-specific Hsp60 were measured by enzyme immunoassay. Human Hsp60 IgA, but not IgG or C pneumoniae Hsp60, antibodies were a significant risk factor for coronary events (odds ratio 2.0, 95% CI 1.1 to 3.6, when the fourth and first quartiles are compared). When an elevated human Hsp60 IgA antibody level (above the second quartile) was present simultaneously with a high C pneumoniae IgA antibody level (the third quartile) and an elevated C-reactive protein level (the second quartile), compared with all factors at low levels, the risk was 7.0 (95% CI 2.6 to 19.1) without adjustment and 5.0 (95% CI 1.8 to 14.2) when adjustment was made for age and smoking. In conclusion, an elevated human Hsp60 IgA antibody level was a risk factor for coronary events, especially when it was present together with C pneumoniae infection and inflammation. PMID- 11884287 TI - Greater oxidative stress in healthy young men compared with premenopausal women. AB - Coronary risk factors, including age, hypertension, diabetes mellitus, hyperlipidemia, and smoking, are associated with enhanced oxidative stress, which is implicated as a potential mechanism for atherogenesis and atherosclerotic cardiovascular diseases. Male sex is one of the well-known cardiovascular risk factors. We tested the hypothesis that oxidative stress is greater in men than in women. Plasma thiobarbituric acid-reactive substances (TBARS) and urinary 8 isoprostaglandin F2alpha (8-iso-PGF2alpha) were measured in 52 young men and 51 age-matched women. The subjects were healthy, were not smokers, and were not taking any medications or vitamins. Age, blood pressure, plasma cholesterol, and glucose did not differ between the groups. Baseline TBARS (2.32 +/- 0.11 [men] versus 1.87 +/- 0.09 [women] nmol/mL, P<0.01) and 8-iso-PGF2alpha (292 +/- 56 [men] versus 164 +/- 25 [women] pg/mg creatinine, P<0.05) were higher in men than in women. Supplementation of antioxidant vitamins for 4 weeks in men produced a significant reduction in TBARS and 8-iso-PGF2alpha by 34% (P<0.01) and 48% (P<0.05), respectively. Plasma superoxide dismutase, catalase, and vitamin E levels were comparable between the groups. Enhanced oxidative stress in men may provide a biochemical link between male sex and atherosclerotic diseases related to oxidative stress. PMID- 11884288 TI - Leukotriene B4 receptor antagonism reduces monocytic foam cells in mice. AB - Leukotriene B4 (LTB4) is a potent chemotactic agent that activates monocytes through the LTB4 receptor (BLTR). We tested the hypothesis that LTB4 receptor blockade would slow atherosclerotic progression by inhibiting monocyte recruitment. Homozygous low-density receptor knockout (LDLr(-/-)) mice and apolipoprotein E deficient (apoE(-/-)) mice were treated with a specific LTB4 receptor antagonist, CP-105,696, for 35 days. In apoE(-/-)mice, treatment with the LTB4 antagonist did not affect plasma lipid concentrations but significantly reduced CD11b levels both in vascular lesions and whole blood. Compared with age matched controls, lipid accumulation and monocyte infiltration were significantly reduced in treated apoE(-/-) mice at all time points tested. Lesion area reduction was also demonstrated in LDLr(-/-) mice maintained on a high-fat diet. LTB4 antagonism had no significant effect on lesion size in mice possessing the null alleles for another chemotactic agent, monocyte chemoattractant protein-1 (MCP-1(-/-)xapoE(-/-)), suggesting MCP-1 and LTB4 may either interact or exert their effects by a common mechanism. These results demonstrate that in a preclinical model of atherosclerosis LTB4 receptor blockade reduces lesion progression and further suggest a previously unrecognized role for LTB4 or other oxidized lipids recognized by the BLTR receptor in the pathogenesis of this disease. PMID- 11884289 TI - Both apolipoprotein E and immune deficiency exacerbate neointimal hyperplasia after vascular injury in mice. AB - In this study, we investigated the role of T and B lymphocytes in neointimal hyperplasia after endothelial denudation. Catheter-induced endothelial denudation of wild-type mice resulted in rapid infiltration of lymphocytes to the site of injury. Mice defective in recombination-activating gene 2 (RAG2(-/-)) showed increased neointimal formation 14 days after vascular injury in comparison to their wild-type immune-competent littermates. Immunohistochemical studies revealed the preponderance of smooth muscle cells and a significantly higher number of proliferating cells in the neointima of the RAG2(-/-) mice. The neointima size and the number of proliferating smooth muscle cells in the injured vessel of RAG2(-/-) mice were similar to those observed in the injured arteries of apolipoprotein E (apoE)-deficient (apoE(-/-)) mice. Interestingly, mice with double apoE and RAG2 gene mutations (apoE(-/-) RAG2(-/-)) displayed similar neointimal characteristics as mice with a single gene defect, suggesting a similar mechanism for apoE and lymphocyte protection against injury-induced neointimal formation. The protective role of lymphocytes against neointimal formation after vascular injury directly contrasts to their reported role in the promotion of atherosclerosis, which was observed in both apoE(+/+) and apoE(-/-) mice. Thus, these results support the hypothesis of different etiology between hyperlipidemia-induced atherosclerosis and injury-induced vascular occlusion. PMID- 11884290 TI - Interleukin-4 deficiency decreases atherosclerotic lesion formation in a site specific manner in female LDL receptor-/- mice. AB - Activated lymphocytes and mast cells have been detected in human atherosclerotic lesions. Interleukin-4 (IL-4) is a prominent cytokine released during the activation of both these cell types, and its mRNA has been detected in human and mouse atherosclerotic lesions. To define the effects of IL-4 on atherogenesis, bone marrow stem cells from either IL-4-/- or IL-4+/+ mice were transplanted into lethally irradiated female low density lipoprotein (LDL) receptor-/- mice. After an interval sufficient to allow engraftment, mice were placed on a diet containing 21% saturated fat, 1.25% cholesterol, and 0.5% cholate. Hematopoietic engraftment was confirmed by the presence of the LDL receptor gene in bone marrow cells. The effect on IL-4 depletion was confirmed by quantifying cytokine release from splenocytes of reconstituted mice. The deficiency of IL-4 in bone marrow derived cells had no effect on serum cholesterol concentrations or on the distribution of cholesterol among lipoproteins. Atherosclerotic lesion formation was not changed in the aortic root. However, deficiency of IL-4 led to reduced lesion size in the arch (9.1 +/- 1.1% versus 2.8 +/- 0.8% of intimal area, P<0.001) and the thoracic aorta (1.2 +/- 0.2% versus 0.4 +/- 0.1%, P<0.002). Therefore, IL-4 deficiency reduced atherosclerotic lesion formation in a site specific manner in female LDL receptor-/- mice fed a high-fat diet. PMID- 11884291 TI - Accumulation of biglycan and perlecan, but not versican, in lesions of murine models of atherosclerosis. AB - Proteoglycan accumulation within the arterial intima has been implicated in lipoprotein retention and in atherosclerosis progression in humans. Two commonly studied murine models of atherosclerosis, the apolipoprotein E (apoE)-deficient (apoE-/-) mouse and the low density lipoprotein receptor-deficient (LDLR-/-) mouse, develop arterial lesions similar to those of human atherosclerosis. However, specific proteoglycan classes that accumulate in lesions of these mice and their relation to the retention of specific apolipoproteins have not been previously determined. In this report, we characterized the distribution of proteoglycans (versican, biglycan, and perlecan) and apolipoproteins (apoB, apoA I, and apoE) in proximal aortic lesions of chow-fed apoE-/- and LDLR-/- mice at 10, 52, and 73 weeks of age. We observed that similar to the apoE-/- mice, the LDLR-/- mice develop intermediate and advanced plaques within 52 weeks of age. Perlecan and biglycan (both are proteoglycans) appeared early in lesion development with distinct expression patterns as the plaques advanced. Versican, a major proteoglycan detected in human plaques, was mostly absent in both strains. ApoA-I and apoB were detected in early through advanced lesions in regions of proteoglycan accumulation in both strains. Our results indicate that proteoglycans may contribute to the retention of lipoproteins at the earliest stage of atherosclerosis in murine models of atherosclerosis. PMID- 11884292 TI - Aortic constriction exacerbates atherosclerosis and induces cardiac dysfunction in mice lacking apolipoprotein E. AB - Despite considerable evidence suggesting that hypertension contributes to the development and progression of atherosclerosis, the causative links remain unclear. We have tested the effects of chronic hypertension induced by suprarenal aortic constriction on the development of atherosclerosis in apolipoprotein E deficient (Apoe-/-) mice. Compared with a sham operation, narrowing the aortic luminal diameter by 33% increased blood pressure proximal to the constriction by approximately 15 mm Hg, but the pressures distal to the constriction were unchanged. Kidney renin mRNA and plasma renin activity were also unaffected. Compared with plaque size after the sham operation, atherosclerotic plaque size in the aortic root 8 weeks after coarctation was increased to 245% and 152% in males and females, respectively. Aortic segments at the constriction were free of atherosclerotic deposits, but segments proximal to the constriction were dilated and had atherosclerotic lesions. Thrombi were present immediately below the constriction in Apoe-/- and wild-type vessels. Surprisingly, compared with wild type mice, the Apoe-/- mice were more susceptible to the cardiac hypertrophy and dysfunction induced by pressure overload. Thus, aortic coarctation exacerbates atherosclerosis in vessels proximal to the constriction without a concomitant increase in the renin-angiotensin system. Our study also suggests that apolipoprotein E plays an important role in modulating cardiac hypertrophy. PMID- 11884293 TI - Hepatic fatty acid synthesis is suppressed in mice with fatty livers due to targeted apolipoprotein B38.9 mutation. AB - Humans and genetically engineered mice with hypobetalipoproteinemia due to truncation-producing mutations of the apolipoprotein B (apoB) gene frequently have fatty livers, because the apoB defect impairs the capacity of livers to export triglycerides (TGs). We assessed the adaptation of hepatic lipid metabolism in our apoB-38.9-bearing mice. Hepatic TG contents were 2- and 4-fold higher in heterozygous and homozygous mice, respectively, compared with wild-type mice. Respective in vivo hepatic fatty acid synthetic rates were reduced to 40% and 15% of the wild-type rate. Hepatic mRNAs for sterol regulatory element binding protein (SREBP)-1c, fatty acid synthase (FAS), and stearoyl coenzyme A desaturase-1 were coordinately decreased. FAS and SREBP-1c mRNA levels were strongly and positively correlated with each other and inversely correlated with hepatic TGs, suggesting that impaired TG export is a potent inhibitor of fatty acid synthesis. In contrast, levels of plasma beta-hydroxybutyrate and of hepatic carnitine palmitoyl transferase and peroxisome proliferator-activated receptor alpha mRNAs were not altered, implying that beta-oxidation was not affected. Fasting followed by refeeding increased hepatic fatty acid synthesis 56-fold over fasting in normal and heterozygous mice but only 24-fold in homozygous mice. Parallel changes occurred in FAS and SREBP-1c mRNAs. Thus, impairment of very low density lipoprotein export downregulates hepatic fatty acid synthesis, but the adaptation is incomplete, resulting in fatty livers. The signals mediating suppression of FAS and SREBP-1c levels remain to be identified. PMID- 11884294 TI - Lipolytically modified triglyceride-enriched HDLs are rapidly cleared from the circulation. AB - The precise biochemical mechanisms underlying the reduction of HDL levels in hypertriglyceridemic states are currently not known. In humans, we showed that triglyceride (TG) enrichment of HDL, as occurs in hypertriglyceridemic states, enhances the clearance of HDL-associated apolipoprotein A-I (apoA-I) from the circulation. In the New Zealand White rabbit (an animal model naturally deficient in hepatic lipase [HL]), however, TG enrichment of HDL is not sufficient to alter the clearance of either the protein or lipid moieties of HDL. In the present study, therefore, we determined in the New Zealand White rabbit the combined effects of ex vivo TG enrichment and lipolytic transformation of HDL by HL on the subsequent metabolic clearance of HDL apoA-I. Results of the in vivo kinetic studies (n=18 animals) showed that apoA-I associated with TG-enriched rabbit HDL modified ex vivo by catalytically active HL was cleared 22% more rapidly versus TG-enriched HDL incubated with heat-inactivated HL, and 26% more rapidly than fasting (TG-poor) HDL incubated with active HL (P<0.05 for both). Furthermore, a strong correlation was observed between the HDL TG content and apoA-I fractional catabolic rate (0.59, P<0.05) in the combined active HL groups. These data establish that TG enrichment of HDL with subsequent lipolysis by HL enhances HDL apoA-I clearance, but neither TG enrichment of HDL without HL lipolysis nor HL lipolysis in the absence of previous TG enrichment of HDL is sufficient to enhance HDL clearance. These data further support the important interaction between HDL TG enrichment and HL action in the pathogenesis of HDL lowering in hypertriglyceridemic states. PMID- 11884295 TI - Total homocysteine lowering treatment among coronary artery disease patients in the era of folic acid-fortified cereal grain flour. AB - The prevalence of deficient plasma folate status and elevated total plasma levels of homocysteine (tHcy), have been dramatically reduced after fortification of all enriched cereal grain flour products with folic acid at 140 microg/100 g flour. Against this new background fortification, we evaluated the tHcy-lowering efficacy of pharmacological dose, folic acid-based vitamin B supplementation among stable coronary artery disease (CAD) patients. Using a 2x2 factorial design, 131 stable CAD patients (mean age 60.1 years; 29.8% women) were randomly assigned to receive a combination of folic acid 2.5 mg/d, riboflavin 5 mg/d, + B12 0.4 mg/d, or placebo, with or without vitamin B6 50 mg/d, for 12 weeks of treatment. ANCOVA adjusted for baseline fasting tHcy levels revealed only very modest (ie, approximately 1.0 micromol/L), albeit statistically significant (P<0.05), reductions in mean fasting tHcy levels afforded by the folic acid containing treatments. Additional analyses indicated that none of the treatments provided a statistically significant reduction in the 2-hour post-methionine increase in tHcy levels, relative to placebo treatment. CAD patients exposed to cereal grain flour products fortified with folic acid who receive high-dose, folic acid-containing vitamin B regimens, experience only very modest reductions in their mean fasting plasma tHcy levels. These findings have important implications for the statistical power of clinical trials testing the hypothesis that tHcy-lowering treatment may reduce recurrent atherothrombotic event rates. PMID- 11884296 TI - Impact of dietary intervention, sex, and apolipoprotein E phenotype on tracking of serum lipids and apolipoproteins in 1- to 5-year-old children: the Special Turku Coronary Risk Factor Intervention Project (STRIP). AB - The effects of dietary intervention, sex, and apolipoprotein E phenotype on tracking of serum lipid values in young children have remained poorly characterized. We investigated these associations in 1062 infants who were randomized into control and intervention groups (n=522 and n=540, respectively) at age 7 months; the intervention group received counseling aimed at maintaining a low-saturated fat, low-cholesterol diet. In 519 children in the control (n=254) and intervention (n=265) groups, serum lipid values were studied annually between 13 months and 5 years of age. In all children, tracking was strongest for the ratio of high density lipoprotein (HDL) cholesterol to total cholesterol; when a 13-month-old child belonged to the lowest quartile of the distribution, the odds ratio for belonging to the same quartile at older ages was 39.0 (95% CI 23.1 to 66.0). Dietary intervention did not influence the tracking of serum lipids. Tracking of HDL cholesterol was stronger in the boys than in the girls (P=0.018). Tracking of non-HDL cholesterol and apolipoprotein B in the children with phenotypes E2/3 or E3/3 was stronger than that in the other children (P=0.031 and P=0.014, respectively). In conclusion, the apolipoprotein E phenotype strongly influences tracking of non-HDL cholesterol and apolipoprotein B values in early childhood, whereas dietary intervention had no effect on tracking of any of the lipids. A child's sex influenced tracking only of HDL cholesterol, with boys showing stronger tracking. PMID- 11884297 TI - Lack of plasminogen activator inhibitor-1 promotes growth and abnormal matrix remodeling of advanced atherosclerotic plaques in apolipoprotein E-deficient mice. AB - Epidemiological studies suggest that elevated plasma levels of plasminogen activator inhibitor-1 (PAI-1) predispose an individual to ischemic heart disease or promote plaque progression by inhibiting fibrinolysis. In the present study, loss of PAI-1 in apolipoprotein E (apoE)-deficient (apoE(-/-):PAI-1(-/-)) mice promoted the growth of advanced atherosclerotic plaques, which was due to enhanced extracellular matrix deposition. ApoE(-/-):PAI-1(-/-) plaques also exhibited collagen fiber disorganization and degradation. Immunostaining and bone marrow transplantation revealed that smooth muscle cells, not macrophages, primarily expressed PAI-1 in plaques. Thus, although PAI-1 may promote plaque growth because of its antifibrinolytic properties, the present study reveals a protective role for PAI-1 by limiting plaque growth and preventing abnormal matrix remodeling. PMID- 11884298 TI - Genetic contribution to circulating levels of hemostatic factors in healthy families with effects of known genetic polymorphisms on heritability. AB - Levels of fibrinogen, factor VII (FVII), factor XIII (FXIII), plasminogen activator inhibitor (PAI)-1, and tissue plasminogen activator have been associated with coronary artery disease as have genetic polymorphisms. Quantitative genetic analyses allow determination of the genetic contribution to phenotypic variation. We investigated familial influences on these hemostatic factors in 537 adults from 89 randomly ascertained healthy families of white North European origin. We used maximum likelihood analysis to estimate the heritabilities of these factors and effects of covariates on the factors in these families. After adjustment for age and sex, the factors showed considerable heritability, varying from 26% (PAI-1) to 47% (FXIII complex). The influence of known polymorphisms was negligible for fibrinogen and contributed 2% to the variance of the FXIII complex and PAI-1 and 11% to the variance of FVII coagulant activity. Age, sex, body mass index, lifestyle, and metabolic covariates explained between 10% (FXIII) and 44% (PAI-1) of phenotypic variance. Childhood household influences significantly affected FVII (11%) and FXIII (18%). A significant degree of phenotypic variance of several hemostatic factors can be explained by additive genes and known covariates. The impact of certain well characterized polymorphisms to the heritability is small in this population of healthy families, indicating the need to localize new genes influencing hemostatic factor levels. PMID- 11884299 TI - Phospholipid-binding domain of factor VIII is involved in endothelial cell mediated activation of factor X by factor IXa. AB - Apparently quiescent, nonapoptotic endothelial cells mediate the activation of factor X by activated factor IX in the presence of its cofactor, activated factor VIII. In a previous study, we reported that during the activation of factor X, the interaction of the cofactor with the endothelial cell membrane clearly differs from the interaction of the cofactor with artificial lipid membranes. In the present study, we identified the peptide domain of factor VIII involved in the assembly of the enzyme-cofactor complex on the endothelial cell surface. With the use of monoclonal antibodies against different peptide sequences on the factor VIII light chain, it was observed that the lipid-binding region of the C2 domain on the factor VIII light chain mediates the assembly of the factor X activating complex on the endothelial cell surface. In addition, a synthetic peptide that constitutes region Ala2318-Tyr2332 of the C2 domain and that is known for its ability to inhibit the binding of factor VIII to artificial lipid membranes also showed inhibition of the cofactor activity of factor VIII on endothelial cells. Thus, the carboxy-terminal part of the factor VIII light chain not only contains sites involved in lipid binding but also contains sites involved in complex assembly on the endothelial cell membrane. PMID- 11884300 TI - A human antibody that inhibits factor IX/IXa function potently inhibits arterial thrombosis without increasing bleeding. AB - 10C12, a human antibody F(ab')2, which specifically binds to the gamma carboxyglutamic acid domain of factor IX/factor IXa (F.IX/IXa), interferes with all known coagulation processes in which F.IX/IXa is involved. In a rabbit model of carotid artery injury, intravenous administration of 10C12 or heparin decreased thrombosis dose dependently. The dose that resulted in a 90% reduction of thrombus mass (ED90) was a 30-microg/kg bolus of 10C12 or a 100-U/kg bolus plus 1.0 U x kg(-1) x min(-1) infusion of heparin. Heparin, at and below the ED90, significantly prolonged coagulation times and cuticle bleeding times. In contrast, 10C12 had no effect on coagulation or bleeding times at doses up to 4 times the ED90. To further evaluate the effect of 10C12 on bleeding, it was compared with heparin in a novel model of blood loss. At the ED90 of heparin, blood loss induced by a standardized injury to the vasculature of the rabbit tibia increased to more than 2 times that of saline controls. In contrast, the dose of 10C12 required to produce a similar increase in blood loss was more than 30 times the ED90. The antithrombotic potency and relative safety of this fully human antibody suggests that it may have therapeutic value for treatment of thrombotic disorders. PMID- 11884301 TI - From patients to end users. PMID- 11884302 TI - Against internet exceptionalism. PMID- 11884303 TI - The quality of health information on the internet. PMID- 11884305 TI - Three new initiatives involving bmj.com. PMID- 11884304 TI - NHS Direct audited. PMID- 11884306 TI - Claim launched against makers of third generation pill. PMID- 11884309 TI - United Kingdom grants first human embryo research licences. PMID- 11884310 TI - Veterans sue ministry of defence over post-traumatic stress disorder. PMID- 11884311 TI - Drugs industry urged to develop medicines for children. PMID- 11884312 TI - Overseas talent can help us build a better NHS, says Magdi Yacoub. PMID- 11884313 TI - "Buyer beware" remains US policy towards information on the net. PMID- 11884314 TI - WHO calls for a health domain name to help consumers. PMID- 11884315 TI - UK government aims to integrate health information on the internet. PMID- 11884316 TI - European Commission to publish a code of practice for websites. PMID- 11884317 TI - Barcelona medical college runs certification system for websites. PMID- 11884318 TI - Trust mark launched as a guarantee of safety in the Netherlands. PMID- 11884319 TI - NHS Direct Online explores partnerships with other health organisations. PMID- 11884320 TI - Examination of instruments used to rate quality of health information on the internet: chronicle of a voyage with an unclear destination. AB - OBJECTIVE: This study updates work published in 1998, which found that of 47 rating instruments appearing on websites offering health information, 14 described how they were developed, five provided instructions for use, and none reported the interobserver reliability and construct validity of the measurements. DESIGN: All rating instrument sites noted in the original study were visited to ascertain whether they were still operating. New rating instruments were identified by duplicating and enhancing the comprehensive search of the internet and the medical and information science literature used in the previous study. Eligible instruments were evaluated as in the original study. RESULTS: 98 instruments used to assess the quality of websites in the past five years were identified. Many of the rating instruments identified in the original study were no longer available. Of 51 newly identified rating instruments, only five provided some information by which they could be evaluated. As with the six sites identified in the original study that remained available, none of these five instruments seemed to have been validated. CONCLUSIONS: Many incompletely developed rating instruments continue to appear on websites providing health information, even when the organisations that gave rise to those instruments no longer exist. Many researchers, organisations, and website developers are exploring alternative ways of helping people to find and use high quality information available on the internet. Whether they are needed or sustainable and whether they make a difference remain to be shown. PMID- 11884321 TI - How do consumers search for and appraise health information on the world wide web? Qualitative study using focus groups, usability tests, and in-depth interviews. AB - OBJECTIVES: To describe techniques for retrieval and appraisal used by consumers when they search for health information on the internet. DESIGN: Qualitative study using focus groups, naturalistic observation of consumers searching the world wide web in a usability laboratory, and in-depth interviews. PARTICIPANTS: A total of 21 users of the internet participated in three focus group sessions. 17 participants were given a series of health questions and observed in a usability laboratory setting while retrieving health information from the web; this was followed by in-depth interviews. SETTING: Heidelberg, Germany. RESULTS: Although their search technique was often suboptimal, internet users successfully found health information to answer questions in an average of 5 minutes 42 seconds (median 4 minutes 18 seconds) per question. Participants in focus groups said that when assessing the credibility of a website they primarily looked for the source, a professional design, a scientific or official touch, language, and ease of use. However, in the observational study, no participants checked any "about us" sections of websites, disclaimers, or disclosure statements. In the post-search interviews, it emerged that very few participants had noticed and remembered which websites they had retrieved information from. CONCLUSIONS: Further observational studies are needed to design and evaluate educational and technological innovations for guiding consumers to high quality health information on the web. PMID- 11884322 TI - Breast cancer on the world wide web: cross sectional survey of quality of information and popularity of websites. AB - OBJECTIVES: To determine the characteristics of popular breast cancer related websites and whether more popular sites are of higher quality. DESIGN: The search engine Google was used to generate a list of websites about breast cancer. Google ranks search results by measures of link popularity---the number of links to a site from other sites. The top 200 sites returned in response to the query "breast cancer" were divided into "more popular" and "less popular" subgroups by three different measures of link popularity: Google rank and number of links reported independently by Google and by AltaVista (another search engine). MAIN OUTCOME MEASURES: Type and quality of content. RESULTS: More popular sites according to Google rank were more likely than less popular ones to contain information on ongoing clinical trials (27% v 12%, P=0.01 ), results of trials (12% v 3%, P=0.02), and opportunities for psychosocial adjustment (48% v 23%, P<0.01). These characteristics were also associated with higher number of links as reported by Google and AltaVista. More popular sites by number of linking sites were also more likely to provide updates on other breast cancer research, information on legislation and advocacy, and a message board service. Measures of quality such as display of authorship, attribution or references, currency of information, and disclosure did not differ between groups. CONCLUSIONS: Popularity of websites is associated with type rather than quality of content. Sites that include content correlated with popularity may best meet the public's desire for information about breast cancer. PMID- 11884323 TI - Accuracy of information on apparently credible websites: survey of five common health topics. PMID- 11884324 TI - Follow up of quality of public oriented health information on the world wide web: systematic re-evaluation. PMID- 11884325 TI - Enhancing public safety in primary care. PMID- 11884326 TI - Science, medicine, and the future: Microdialysis. PMID- 11884327 TI - Lesson of the week: Immobilisation of the cervical spine in children. PMID- 11884328 TI - ABC of clinical electrocardiography: Atrial arrhythmias. PMID- 11884329 TI - How to find the good and avoid the bad or ugly: a short guide to tools for rating quality of health information on the internet. PMID- 11884330 TI - Education and debate: Regulating health information: a US perspective. PMID- 11884331 TI - Statistics Notes: Validating scales and indexes. PMID- 11884332 TI - Effectiveness of smoking cessation initiatives. Efforts must take into account smokers' disillusionment with smoking and their delusions about stopping. PMID- 11884333 TI - Commentary: Iloprost for cholesterol emboli syndrome. PMID- 11884334 TI - Deaths from chickenpox. Deaths from chickenpox in adults are decreasing. PMID- 11884335 TI - Secondary prevention of coronary heart disease. Ill defined inclusion criteria resulted in missed trials. PMID- 11884336 TI - Advances in virtual reality are wide ranging. PMID- 11884337 TI - News article on report about drug researcher was biased. PMID- 11884338 TI - Dysphagia. Causes of high dysphagia should be assessed by ENT surgeons. PMID- 11884339 TI - Psychiatrists' perspective is insufficient to root out racism. PMID- 11884340 TI - Management of Helicobacter pylori infection. Treatment of ulcers can be improved and over-reliance on proton pump inhibitors reduced. PMID- 11884341 TI - Clinical databases can complement controlled trials. PMID- 11884342 TI - Hospital revises its own data in government league table. PMID- 11884343 TI - Death rates of surgeons should not be published. PMID- 11884349 TI - Large-scale neural model for visual attention: integration of experimental single cell and fMRI data. AB - A computational neuroscience framework is proposed to better understand the role and the neuronal correlate of spatial attention modulation in visual perception. The model consists of several interconnected modules that can be related to the different areas of the dorsal and ventral paths of the visual cortex. Competitive neural interactions are implemented at both microscopic and interareal levels, according to the biased competition hypothesis. This hypothesis has been experimentally confirmed in studies in humans using functional magnetic resonance imaging (fMRI) techniques and also in single-cell recording studies in monkeys. Within this neuro-dynamical approach, numerical simulations are carried out that describe both the fMRI and the electrophysiological data. The proposed model draws together data of different spatial and temporal resolution, as are the above-mentioned imaging and single-cell results. PMID- 11884351 TI - Directing attention to locations and to sensory modalities: multiple levels of selective processing revealed with PET. AB - We used positron emission tomography (PET) to investigate the neural correlates of selective attention in humans. We examined the effects of attending to one side of space versus another (spatial selection) and to one sensory modality versus another (intermodal selection) during bilateral, bimodal stimulation of vision and touch. Attention toward one side resulted in greater activity in several contralateral areas. In somatosensory cortex, these spatial attentional modulations were found only when touch was relevant. In the intraparietal sulcus, spatial attentional effects were multimodal, independent of the modality attended. In occipital areas, spatial modulations were also found during both visual and tactile attention, indicating that tactile attention can affect activity in visual cortex; but occipital areas also showed more activity overall during visual attention. This suggests that while spatial attention can exert multimodal influences on visual areas, these still maintain their specificity for the visual modality. Additionally, irrespective of the attended side, attending to vision activated posterior parietal and superior premotor cortices, while attending to touch activated the parietal operculi. We conclude that attentional selection operates at multiple levels, with attention to locations and attention to modalities showing distinct effects. These jointly contribute to boost processing of stimuli at the attended location in the relevant modality. PMID- 11884350 TI - Abnormalities of SNARE mechanism proteins in anterior frontal cortex in severe mental illness. AB - A fundamental molecular component of neural connectivity is the SNARE (SNAP receptor) protein complex, which consists of three proteins, syntaxin, SNAP-25 and VAMP. Under appropriate conditions, the SNARE complex can be formed in vitro. To investigate the hypothesis that dysregulation of SNARE proteins or their interactions could be abnormal in severe mental disorders, the three SNARE proteins and the complex were studied in post-mortem anterior frontal cortex homogenates. An ELISA was used to quantify SNARE protein immunoreactivities in cortical homogenates from four groups: patients with schizophrenia who died of causes other than suicide (n = 6), patients with schizophrenia and suicide (n = 7), patients with depression and suicide (n = 11), and controls (n = 11). Differences between groups in patterns of SNARE protein immuno-reactivities were demonstrated [Wilks' Lambda F(9,68) = 3.57, P = 0.001]. Protein-by-protein analyses indicated a significant reduction in SNAP-25 immunoreactivity in the schizophrenia non-suicide group [28% decrease relative to controls, F(3,31) = 6.45, P = 0.002, Student-Newman-Keuls test, P < 0.01]. The intercorrelations between SNARE protein and synaptophysin immunoreactivities were high in controls, but lower in the other groups, further indicating disturbances in relationships between these proteins. The extent of SNARE complex formation in vitro was studied using immuno-blotting. Significant differences related to group membership were observed for the SNARE complexes identified by SNAP-25 [Wilks' Lambda F(3,31) = 4.76, P = 0.008] and by syntaxin immunostaining [Wilks' Lambda F(3,31) = 9.16, P = 0.0002]. In both groups with suicide as a cause of death, relatively more SNAP-25 and syntaxin was present in the heterotrimeric SNARE complex than in other molecular forms. These abnormalities in the SNARE complex could represent a molecular substrate for abnormalities of neural connectivity in severe mental disorders. PMID- 11884352 TI - Segregation of areas related to visual working memory in the prefrontal cortex revealed by rTMS. AB - The functional organization of working memory (WM) in the human prefrontal cortex remains unclear. Storage and processing functions might be segregated in ventral and dorsal areas of the prefrontal cortex, respectively. If so, storage functions might be spared, irrespective of informational domain, following damage or dysfunction in dorsolateral areas. Alternatively, WM and prefrontal function in general might be segregated according to informational domains (e.g. spatial versus object-based information). In the present study we used repetitive transcranial magnetic stimulation (rTMS) to directly test these competing hypotheses. We applied rTMS to transiently and selectively disrupt the function of the dorsomedial, dorsolateral or ventral prefrontal cortex in normal human volunteers performing either a spatial or a face-recognition delayed-response task. Performance in the spatial task was impaired by rTMS of the dorsomedial prefrontal cortex. Performance in the face-recognition (non-spatial) task was impaired by rTMS of the ventral prefrontal cortex. Transient disruption of the dorsolateral prefrontal cortex affected performance in both tasks. These findings provide evidence of domain-specific segregation of WM functions in widely separated areas of prefrontal cortex. PMID- 11884353 TI - Representation of pain and somatic sensation in the human insula: a study of responses to direct electrical cortical stimulation. AB - We studied painful and non-painful somaesthetic sensations elicited by direct electrical stimulations of the insular cortex performed in 43 patients with drug refractory temporal lobe epilepsy, using stereotactically implanted depth electrodes. Painful sensations were evoked in the upper posterior part of the insular cortex in 14 patients, mostly in the right hemisphere. Non-painful sensations were elicited in the posterior part of the insular cortex in 16 patients, in both hemispheres. Thus, painful and non-painful somaesthetic representations in the human insula overlap. Both types of responses showed a trend toward a somatotopic organization. These results agree with previous anatomical and unit recording studies in monkeys indicating a participation of the posterior part of the insular cortex in processing both noxious and innocuous somaesthetic stimuli. In humans, both a posterior and an anterior pain-related cortical area have been described within the insular cortex using functional imaging. Our results help to define the respective functional roles of these two insular areas. Finally, lateralization in the right hemisphere of sites where painful sensations were evoked is coherent with the hypothesis of a preponderant role of this hemisphere in species survival. PMID- 11884354 TI - Reduced neuronal size and glial cell density in area 9 of the dorsolateral prefrontal cortex in subjects with major depressive disorder. AB - Reductions in glial cell density and neuronal size have been described recently in major depressive disorder (MDD). Considering the important trophic influence of glia on neurons, we hypothesized that this glial cell deficit is more prominent close to neurons. In this investigation we have characterized neuronal and glia cytoarchitecture in prefrontal area 9 using spatial point pattern techniques and two-dimensional measures of cell size and density. In post-mortem brain tissue of subjects with MDD, schizophrenia, bipolar disorder (BPD), and normal controls (15 subjects per group), we examined the laminar location and size of all neurons and glial nuclei in a 500 microm wide strip of cortex extending from the pia to the grey-white matter border. In MDD, we observed reductions in glial cell density (30%; P = 0.007) in layer 5 and neuronal size (20%; P = 0.003) in layer 6. We also found that glial cell density (34%; P = 0.003) was reduced in layer 5 in schizophrenia, while neuronal size was reduced in layers 5 (14%) (P = 0.006) and 6 (18%; P = 0.007) in BPD. The spatial pattern investigation of neurons and glia demonstrated no alteration in the clustering of glia about neurons between control and patient groups. These findings confirm that glial cell loss and neuronal size reductions occur in the deeper cortical layers in MDD, but provide no support for the hypothesis that an altered spatial distribution of glia about neurons plays a role in the development of these changes. PMID- 11884355 TI - Anatomical, physiological, molecular and circuit properties of nest basket cells in the developing somatosensory cortex. AB - Anatomical, electrophysiological and molecular diversity of basket cell-like interneurons in layers II-IV of rat somatosensory cortex were studied using patch clamp electrodes filled with biocytin. This multiparametric study shows that neocortical basket cells (BCs) are composed of three distinct subclasses: classical large (LBC) and small (SBC) basket cells and a third subclass, the nest basket cell (NBC). Anatomically, NBCs were distinct from LBCs and SBCs in that they formed simpler dendritic arbors and an axonal plexus of inter-mediate density, composed of a few long, smooth axonal branches. Electrophysiologically, NBCs exhibited diverse discharge responses to depolarizing current injections including accommodation, non-accommodation and stuttering. Single-cell multiplex RT-PCR revealed distinct mRNA expression patterns for the calcium binding proteins parvalbumin (PV), calbindin (CB) and calretinin (CR), and the neuropeptides somatostatin (SOM), vasoactive intestinal peptide (VIP), cholecystokinin (CCK) and neuropeptide Y (NPY) for each BC-subclass. SBCs lacked NPY expression but invariably expressed VIP, whereas neither VIP, CR nor SOM expression was detected in LBCs, and VIP and CR expression was absent in NBCs. Electro-physiologically distinct types of NBCs formed GABAergic synapses with specific dynamics onto pyramidal cells (PCs) and received either strongly facilitating or depressing synaptic inputs from PCs. Finally, NBCs were found to be the most common basket cell in layers II/III, while LBCs were the most common in layer IV. These data provide multiparametric distinguishing features of three major subclasses of basket cells and indicate that NBCs are powerful interneurons that provide most of the (peri-)somatic inhibition in the supragranular layers. PMID- 11884356 TI - Architecture and callosal connections of visual areas 17, 18, 19 and 21 in the ferret (Mustela putorius). AB - Visual areas 17, 18, 19 and 21 of the ferret can be distinguished on the grounds of cytoarchitecture, myeloarchitecture and cytochrome oxidase reactivity, and with transneuronal tract-tracing from the eye. Each visual area contains callosally connected, as well as acallosal, regions. The callosal connections originate mainly from layers 2 and 3 and, more widely, from layer 6. Callosally projecting neurons and callosal terminals are organized in three roughly medio laterally oriented bands. The posterior and intermediate bands straddle the 17/18 and 19/21 border, respectively; the third band extends along the medial bank of the lateral suprasylvian sulcus. These bands are linked by a variable number of bridges of connections that demarcate acallosal islands. The distribution of callosal connections predicts the existence of vertical meridian representations corresponding to each of the bands and of non-isotropic representations of the visual field within the bridges and islands. PMID- 11884357 TI - The representation of the visual field in three extrastriate areas of the ferret (Mustela putorius) and the relationship of retinotopy and field boundaries to callosal connectivity. AB - We describe representations of the visual field in areas 18, 19 and 21 of the ferret using standard microelectrode mapping techniques. In all areas the azimuths are represented as islands of peripheral visual field surrounded by central visual field representation. The zero meridian was found at the 17/18 and 19/21 borders; at the 18/19 and anterior border of 21 the relative periphery of the visual field was found. In areas 18 and 19, elevations are represented in a smooth medio-lateral progression from lower to upper visual field. In several cases the elevations in area 21 evidenced a similar medio-lateral progression; however, in others the elevations exhibited a split representation of the horizontal meridian. Anatomically determined callosal connections coincided with the representation of azimuths near the zero meridian. Medio-lateral bands of callosal connectivity that straddle the 17/18 and 19/21 borders are connected by bridges of callosally projecting cells. Acallosal cortical islands corresponded to the peripheral visual field and were found straddling the 18/19 border and the anterior border of area 21. The results are discussed in relation to callosal connectivity and retinotopy in extrastriate visual cortex and to proposed homologies of carnivore and primate visual cortex. PMID- 11884358 TI - Differential rates of regional brain change in callosal and ventricular size: a 4 year longitudinal MRI study of elderly men. AB - Brain structure changes in size with normal aging, but the rate at which different structures change is controversial. We used magnetic resonance imaging (MRI) performed twice, 4 years apart, to compare rates of age-related size change of the corpus callosum, which has been inconsistently observed to thin with age, with change in the lateral ventricles, which are well established to enlarge. Subjects were 215 community dwelling, elderly men (70-82 years old at initial MRI), who were participants in a longitudinal study of cardiovascular risk factors. Percent change in size was significant for both the callosal and ventricular measures, but annual rate of ventricular expansion (2.9%) was significantly greater than annual rate of callosal thinning (-0.9%). Callosal regions showed statistically equivalent rates of shrinkage; ventricular dilatation was symmetrical. Neither callosal and ventricular rates of change correlated with each other (r = 0.01), nor did genu and splenium rates of change correlate with each other (r = 0.05). Tests of speeded processing were administered contemporaneously with both MRIs to examine functional ramifications of observed brain changes. Decline in the Mini-Mental State Examination was related to thinning of the splenium, and decline in Stroop test word reading was selectively related to thinning of the callosal body. These longitudinal data support the contentions that differential rates of change occur in different brain regions in normal aging, age-related callosal thinning contributes to functional declines, and rate of change in one region can be independent of rate of change in another region, even within a brain structure. PMID- 11884359 TI - Mechanical noxious stimuli cause bilateral activation of parietal operculum in callosotomized subjects. AB - The patterns of cortical activation evoked by tactile and mechanical painful stimulation in six normal subjects and three patients with complete resection of the corpus callosum are described and compared, with emphasis on the parietal operculum. Stimulus-related cortical activation was investigated by functional magnetic resonance imaging. In both groups, painful stimulation activated the first somatosensory, insular and cingulate cortices in the contralateral hemisphere, and the parietal opercular cortex in both hemispheres. Comparison between the two patterns of cortical activation demonstrated that ipsilateral activation by unilateral painful stimulation is at least partially independent of the corpus callosum and suggests a different organization of the pain and touch systems. PMID- 11884361 TI - PAR2 is partout and now in the heart. PMID- 11884362 TI - Gaining respectability: membrane-delimited, caveolar-restricted activation of ion channels. PMID- 11884363 TI - Can integrins integrate vascular myogenic responses? PMID- 11884364 TI - Autocrine stimulation of cardiac Na(+)-Ca(2+) exchanger currents by endogenous endothelin released by angiotensin II. AB - The goal of the present study was to evaluate the effects of Ang II on the current produced by the Na(+)-Ca(2+) exchanger (I(NCX)) working in the reverse mode and the possible autocrine role played by the release of endothelin (ET) in these actions. I(NCX) was studied in isolation in cat cardiac myocytes. Angiotensin II (Ang II) (100 nmol/L) increased I(NCX) at potentials higher than 0 mV (at +60 mV: 2.07 +/- 0.22 pA/pF in control versus 2.73 +/- 0.22 pA/pF in Ang II, n=9; P<0.05). The increase in I(NCX) induced by Ang II was prevented by the treatment of the cells with the unspecific blocker of the ET receptors, TAK 044 (1 micromol/L) (at +60 mV: 2.15 +/- 0.27 pA/pF in control versus 2.01+/- 0.26 pA/pF in Ang II, n=5, NS). These results show, for the first time, that the effect of Ang II on I(NCX) is the result of the autocrine actions of ET released by the octapeptide. PMID- 11884365 TI - Phosphorylation of glycogen synthase kinase-3beta during preconditioning through a phosphatidylinositol-3-kinase--dependent pathway is cardioprotective. AB - We previously reported that activation of phosphatidylinositol-3-kinase (PI3 kinase) is involved in ischemic preconditioning (PC). Our goal was to determine downstream targets of PI3-kinase. In perfused rat hearts, PC (4 cycles of 5 minutes of ischemia and 5 minutes of reflow) increased phosphorylation of glycogen synthase kinase-3beta (GSK-3beta), a downstream target of PI3-kinase and protein kinase B (PKB), an effect that was blocked by wortmannin. Because phosphorylation inactivates GSK-3beta, we examined whether PC-induced phosphorylation and inhibition of GSK-3beta is important in PC by using two inhibitors of GSK-3beta, lithium and SB 216763. Pretreatment of perfused rat hearts with lithium or SB 216763, before ischemia, mimicked the protective effects of PC; hearts treated with either lithium or SB 216763 had improved postischemic function and reduced infarct size. These findings indicate that inhibition of GSK-3beta is protective and that this PI3-kinase--dependent signaling pathway may play an important role in ischemic preconditioning. PMID- 11884366 TI - Applied proteomics: mitochondrial proteins and effect on function. AB - The identification of a majority of the polypeptides in mitochondria would be invaluable because they play crucial and diverse roles in many cellular processes and diseases. The endogenous production of reactive oxygen species (ROS) is a major limiter of life as illustrated by studies in which the transgenic overexpression in invertebrates of catalytic antioxidant enzymes results in increased lifespans. Mitochondria have received considerable attention as a principal source---and target---of ROS. Mitochondrial oxidative stress has been implicated in heart disease including myocardial preconditioning, ischemia/reperfusion, and other pathologies. In addition, oxidative stress in the mitochondria is associated with the pathogenesis of Alzheimer's disease, Parkinson's disease, prion diseases, and amyotrophic lateral sclerosis (ALS) as well as aging itself. The rapidly emerging field of proteomics can provide powerful strategies for the characterization of mitochondrial proteins. Current approaches to mitochondrial proteomics include the creation of detailed catalogues of the protein components in a single sample or the identification of differentially expressed proteins in diseased or physiologically altered samples versus a reference control. It is clear that for any proteomics approach prefractionation of complex protein mixtures is essential to facilitate the identification of low-abundance proteins because the dynamic range of protein abundance within cells has been estimated to be as high as 10(7). The opportunities for identification of proteins directly involved in diseases associated with or caused by mitochondrial dysfunction are compelling. Future efforts will focus on linking genomic array information to actual protein levels in mitochondria. PMID- 11884367 TI - Mitochondrial PKCepsilon and MAPK form signaling modules in the murine heart: enhanced mitochondrial PKCepsilon-MAPK interactions and differential MAPK activation in PKCepsilon-induced cardioprotection. AB - Although activation of protein kinase C (PKC) epsilon and mitogen-activated protein kinases (MAPKs) are known to play crucial roles in the manifestation of cardioprotection, the spatial organization of PKCepsilon signaling modules in naive and protected myocardium remains unknown. Based on evidence that mitochondria are key mediators of the cardioprotective signal, we hypothesized that PKCepsilon and MAPKs interact, and that they form functional signaling modules in mitochondria during cardioprotection. Both immunoblotting and immunofluorescent staining demonstrated that PKCepsilon, ERKs, JNKs, and p38 MAPK co-localized with cardiac mitochondria. Moreover, transgenic activation of PKCepsilon greatly increased mitochondrial PKCepsilon expression and activity, which was concomitant with increased mitochondrial interaction of PKCepsilon with ERKs, JNKs, and p38 as determined by co-immunoprecipitation. These complex formations appeared to be independent of PKCepsilon activity, as the interactions were also observed in mice expressing inactive PKCepsilon. However, although both active and inactive PKCepsilon bound to all three MAPKs, increased phosphorylation of mitochondrial ERKs was only observed in mice expressing active PKCepsilon but not in mice expressing inactive PKCepsilon. Examination of potential downstream targets of mitochondrial PKCepsilon-ERK signaling modules revealed that phosphorylation of the pro-apoptotic protein Bad was elevated in mitochondria. Together, these data show that PKCepsilon forms subcellular targeted signaling modules with ERKs, leading to the activation of mitochondrial ERKs. Furthermore, formation of mitochondrial PKCepsilon-ERK modules appears to play a role in PKCepsilon-mediated cardioprotection, in part by the phosphorylation and inactivation of Bad. PMID- 11884368 TI - Human vascular smooth muscle cells from restenosis or in-stent stenosis sites demonstrate enhanced responses to p53: implications for brachytherapy and drug treatment for restenosis. AB - The p53 tumor suppressor gene regulates growth arrest and apoptosis after DNA damage. Recent studies suggest that p53 is inactive in vascular smooth muscle cells (VSMCs) in human angioplasty restenosis, promoting VSMC accumulation and vessel stenosis. In contrast, the success of irradiation (brachytherapy) for in stent restenosis argues that DNA-damage p53 responses are intact. We examined p53 expression and function in human VSMCs from normal vessels (n-VSMCs) and angioplasty/in-stent restenosis sites (r-VSMCs). p53 expression was uniformly low in all VSMCs and was induced by DNA damage. However, p53 induced profoundly different biological effects in r-VSMCs versus n-VSMCs, causing growth arrest and apoptosis in r-VSMCs only. In addition, dominant-negative p53 promoted cell proliferation and apoptosis in r-VSMCs but not n-VSMCs. Cytotoxic drug-- or irradiation-induced growth arrest and apoptosis in both cell types was mediated only partly by p53. In contrast, cyclin D degradation in response to DNA damage, a critical early mediator of growth arrest, was impaired in r-VSMCs, an effect that required p53. We conclude that p53 expression and function are normal or increased in r-VSMCs and may underlie the success of brachytherapy. We also identify a restenosis VSMC-specific defect in cyclin D degradation induced by DNA damage. PMID- 11884369 TI - Sp1 transcription factor as a molecular target for nitric oxide-- and cyclic nucleotide--mediated suppression of cGMP-dependent protein kinase-Ialpha expression in vascular smooth muscle cells. AB - cGMP-dependent protein kinase (PKG) expression is highly variable and decreases in cultured vascular smooth muscle cells (VSMCs), exposure of cells to nitric oxide (NO), or in response to balloon catheter injury in vivo. In this study, the mechanisms of human type I PKG-alpha (PKG-Ialpha) gene expression were examined. Three structurally unrelated NO donors decreased PKG-Ialpha promoter activity after transfection of a promoter/luciferase construct in VSMCs. Promoter deletion analysis demonstrated that (1) a 120-bp promoter containing tandem Sp1 sites was sufficient to drive basal PKG-Ialpha promoter activity, and (2) NO was inhibitory at this site. Cyclic nucleotide analogues also suppressed PKG-Ialpha promoter activity with cAMP being more potent than cGMP. The effects of cyclic nucleotides to suppress PKG-Ialpha promoter activity were attenuated by a specific cAMP dependent protein kinase (PKA) inhibitor. Single or double mutation of Sp1 binding sites abolished PKG-Ialpha expression. Moreover, Sp1 binding activity on the PKG-Ialpha promoter was detected in A7r5 cells, and this binding was inhibited by NO and cyclic nucleotides. These results indicate that PKG-Ialpha gene expression is driven by an Sp1 transcription mechanism, and that NO and cAMP inhibit Sp1-mediated PKG-Ialpha gene expression through separate mechanisms. PMID- 11884370 TI - Estradiol alters nitric oxide production in the mouse aorta through the alpha-, but not beta-, estrogen receptor. AB - Although estradiol (E(2)) has been recognized to exert several vasculoprotective effects in several species, its effects in mouse vasomotion are unknown, and consequently, so is the estrogen receptor subtype mediating these effects. We investigated the effect of E(2) (80 microg/kg/day for 15 days) on NO production in the thoracic aorta of ovariectomized C57Bl/6 mice compared with those given placebo. E(2) increased basal NO production. In contrast, the relaxation in response to ATP, to the calcium ionophore A23187, and to sodium nitroprusside was unaltered by E(2), whereas acetylcholine-elicited relaxation was decreased. The abundance of NO synthase I, II, and III immunoreactive proteins (using Western blot) in thoracic aorta homogenates was unchanged by E(2). To determine the estrogen receptor (ER) subtype involved in these effects, transgenic mice in which either the ERalpha or ERbeta has been disrupted were ovariectomized and treated, or not, with E(2). Basal NO production was increased and the sensitivity to acetylcholine decreased in ERbeta knockout mice in response to E(2), whereas this effect was abolished in ERalpha knockout mice. Finally, these effects of E(2) on vasomotion required long-term and/or in vivo exposure, as short-term incubation of aortic rings with 10 nmol/L E(2) in the isolated organ chamber did not elicit any vasoactive effects. In conclusion, this study demonstrates that ERalpha, but not ERbeta, mediates the beneficial effect of E(2) on basal NO production. PMID- 11884371 TI - Lovastatin enhances ecto-5'-nucleotidase activity and cell surface expression in endothelial cells: implication of rho-family GTPases. AB - Extracellular adenosine production by the GPI-anchored Ecto-5'-Nucleotidase (Ecto 5'-Nu) plays an important role in the cardiovascular system, notably in defense against hypoxia. It has been previously suggested that HMG-CoA reductase inhibitors (HRIs) could potentiate the hypoxic stimulation of Ecto-5'Nu in myocardial ischemia. In order to elucidate the mechanism of Ecto-5'-Nu stimulation by HRIs, Ecto-5'-Nu activity and expression were determined in an aortic endothelial cell line (SVAREC) incubated with lovastatin. Lovastatin enhanced Ecto-5'-Nu activity in a dose-dependent manner. This increase was not supported by de novo synthesis of the enzyme because neither the mRNA content nor the total amount of the protein were modified by lovastatin. By contrast, lovastatin enhanced cell surface expression of Ecto-5'-Nu and decreased endocytosis of Ecto-5'-Nu, as evidenced by immunostaining. This effect appeared unrelated to modifications of cholesterol content or Ecto-5'-Nu association with detergent-resistant membranes. The effect of lovastatin was reversed by mevalonate, the substrate of HMG-CoA reductase, by its isoprenoid derivative, geranyl-geranyl pyrophosphate, and by cytotoxic necrotizing factor, an activator of Rho-GTPases. Stimulation of Ecto-5'-Nu by lovastatin enhanced the inhibition of platelet aggregation induced by endothelial cells. In conclusion, lovastatin enhances Ecto-5'-Nu activity and membrane expression in endothelial cells. This effect seems independent of lowering cholesterol content but could be supported by an inhibition of Ecto-5'-Nu endocytosis through a decrease of Rho-GTPases isoprenylation. PMID- 11884372 TI - A structural and dynamic investigation of the facilitating effect of glycoprotein IIb/IIIa inhibitors in dissolving platelet-rich clots. AB - Glycoprotein IIb/IIIa (GP IIb/IIIa) inhibitors were shown recently to facilitate the rate and the extent of pharmacological thrombolysis. However, their synergistic potential with rtPA in dissolving thrombotic vaso-occlusions is not fully understood. We have therefore developed a dynamic and structural approach for analysis of fibrinolysis to assess the inhibiting effect of platelets and the facilitating effect of GPIIb/IIIa inhibitors in dissolving platelet-rich clots (PRCs). Fluorescent rtPA was used to study the architecture of PRCs, to follow the progression of the rtPA binding front, and to measure the lysis-front velocity using confocal microscopy. Fibrinolysis resistance of PRCs was related to a reduction of both rtPA binding and lysis-front velocities of platelet-rich areas compared with platelet-poor areas (2.4 +/- 0.2 versus 3.5 +/- 0.4 microm/min for rtPA binding velocity, P=0.04, and 1.2 +/- 0.6 versus 2.8 +/- 0.2 microm/min for lysis-front velocity, P=0.008, in platelet-rich and platelet-poor areas, respectively). Fibrinolysis appeared heterogeneous, leaving platelet-rich areas un-lysed. Adding pharmacological concentrations of abciximab (0.068 micromol/L) or eptifibatide (1 micromol/L) before clotting decreased the average surface of platelet-rich areas by 64% (P=0.0005) and 72% (P=0.0007), respectively. The resulting equalization of rtPA binding rate and rtPA binding front velocity between platelet-rich and platelet-poor areas led to a 3-fold increase of the lysis-front velocity in platelet-rich areas of either abciximab PRC (P=0.006) or eptifibatide-PRC (P=0.03). The overall lysis rate of treated-PRC was increased by 74% compared with control-PRC (P<0.01). These results demonstrate that fibrinolysis resistance of PRCs is related primarily to the heterogeneity in the clot structure between platelet-rich and platelet-poor areas. GP IIb/IIIa inhibitors facilitate the rate and the extent of fibrinolysis by improving rtPA binding velocity and, subsequently, the lysis rate in platelet rich areas. These findings provide new insights on the synergistic potential of GP IIb/IIIa inhibitors and fibrinolytic agents. PMID- 11884373 TI - L-type Ca(2+) currents overlapping threshold Na(+) currents: could they be responsible for the "slip-mode" phenomenon in cardiac myocytes? AB - Phosphorylation of Na channels has been suggested to increase their Ca permeability. Termed "slip-mode conductance" (SMC), this hypothesis predicts that Ca influx via protein kinase A (PKA)-modified Na channels can induce sarcoplasmic reticulum (SR) Ca release. We tested this hypothesis by determining if SR Ca release is graded with I(Na) in the presence of activated PKA (with Isoproterenol, ISO). V(m), I(m), and [Ca](i) were measured in feline (n=26) and failing human (n=19) ventricular myocytes. Voltage steps from -70 through -40 mV were used to grade I(Na). Na channel antagonists (tetrodotoxin), L-type Ca channel (I(Ca,L)) antagonists (nifedipine, cadmium, verapamil), and agonists (Bay K 8644, FPL 64176) were used to separate SMC from I(Ca,L). In the absence of ISO, I(Na) was associated with SR Ca release in human but not feline myocytes. After ISO, graded I(Na) was associated with small amounts of SR Ca release in feline myocytes and the magnitude of release increased in human myocytes. I(Na)-related SR Ca release was insensitive to tetrodotoxin (n=10) but was blocked by nifedipine (n=10) and cadmium (n=3). SR Ca release was induced over the same voltage range in the absence of ISO with Bay K 8644 and FPL 64176 (n=9). Positive voltage steps (to 0 mV) to fully activate Na channels (SMC) in the presence of ISO and Verapamil only caused SR Ca release when block of I(Ca,L) was incomplete. We conclude that PKA-mediated increases in I(Ca,L) and SR Ca loading can reproduce many of the experimental features of SMC. PMID- 11884374 TI - Localization of cardiac sodium channels in caveolin-rich membrane domains: regulation of sodium current amplitude. AB - This study demonstrates that caveolae, omega-shaped membrane invaginations, are involved in cardiac sodium channel regulation by a mechanism involving the alpha subunit of the stimulatory heterotrimeric G-protein, Galpha(s), via stimulation of the cell surface beta-adrenergic receptor. Stimulation of beta-adrenergic receptors with 10 micromol/L isoproterenol in the presence of a protein kinase A inhibitor increased the whole-cell sodium current by a "direct" cAMP-independent G-protein mechanism. The addition of antibodies against caveolin-3 to the cell's cytoplasm via the pipette solution abrogated this direct G protein-induced increase in sodium current, whereas antibodies to caveolin-1 or caveolin-2 did not. Voltage-gated sodium channel proteins were found to associate with caveolin rich membranes obtained by detergent-free buoyant density separation. The purity of the caveolar membrane fraction was verified by Western blot analyses, which indicated that endoplasmic/sarcoplasmic reticulum, endosomal compartments, Golgi apparatus, clathrin-coated vesicles, and sarcolemmal membranes were excluded from the caveolin-rich membrane fraction. Additionally, the sodium channel was found to colocalize with caveolar membranes by immunoprecipitation, indirect immunofluorescence, and immunogold transmission electron microscopy. These results suggest that stimulation of beta-adrenergic receptors, and thereby Galpha(s), promotes the presentation of cardiac sodium channels associated with caveolar membranes to the sarcolemma. PMID- 11884375 TI - Role of the carboxyl terminal of connexin43 in transjunctional fast voltage gating. AB - Previous studies show that chemical regulation of connexin43 (Cx43) gap junction channels depends on the integrity of the carboxyl terminal (CT) domain. Experiments using Xenopus oocytes show that truncation of the CT domain alters the time course for current inactivation; however, correlation with the behavior of single Cx43 channels has been lacking. Furthermore, whereas chemical gating is associated with a "ball-and-chain" mechanism, there is no evidence whether transjunctional voltage regulation for Cx43 follows a similar model. We provide data on the properties of transjunctional currents from voltage-clamped pairs of mammalian tumor cells expressing either wild-type Cx43 or a mutant of Cx43 lacking the carboxyl terminal domain (Cx43M257). Cx43 transjunctional currents showed bi-exponential decay and a residual steady-state conductance of approximately 35% maximum. Transjunctional currents recorded from Cx43M257 channels displayed a single, slower exponential decay. Long transjunctional voltage pulses caused virtual disappearance of the residual current at steady state. Single channel data revealed disappearance of the residual state, increase in the mean open time, and slowing of the transition times between open and closed states. Coexpression of CxM257 with Cx43CT in a separate fragment restored the lower conductance state. We propose that Cx43CT is an effector of fast voltage gating. Truncation of Cx43CT limits channel transitions to those occurring across the higher energy barrier that separates open and closed states. We further propose that a ball-and-chain interaction provides the fast component of voltage-dependent gating between CT domain and a receptor affiliated with the pore. PMID- 11884376 TI - Cardiac myocyte-specific excision of the beta1 integrin gene results in myocardial fibrosis and cardiac failure. AB - Integrins link the extracellular matrix to the cellular cytoskeleton and serve important roles in cell growth, differentiation, migration, and survival. Ablation of beta1 integrin in all murine tissues results in peri-implantation embryonic lethality. To investigate the role of beta1 integrin in the myocardium, we used Cre-LoxP technology to inactivate the beta1 integrin gene exclusively in ventricular cardiac myocytes. Animals with homozygous ventricular myocyte beta1 integrin gene excision were born in appropriate numbers and grew into adulthood. These animals had 18% of control levels of beta1D integrin protein in the heart and displayed myocardial fibrosis. High-fidelity micromanometer-tipped catheterization of the intact 5-week-old beta1 integrin knockout mice showed depressed left ventricular basal and dobutamine-stimulated contractility and relaxation (LV dP/dt(max) and LV dP/dt(min)) as compared with control groups (n=8 to 10 of each, P<0.01). Hemodynamic loading imposed by 7 days of transverse aortic constriction showed that the beta1 integrin knockout mice were intolerant of this stress as they had 53% survival versus 88% in controls (n=15 each). By 6 months of age, mice with depressed ventricular expression of beta1 integrin developed a dilated cardiomyopathy that was not evident in any control animals and had patchy decrease in glucose metabolism as determined by positron emission tomography. Myocyte membrane integrity as determined via Evan's blue dye staining was disrupted in the beta1 integrin knockout mice. This model provides strong evidence for the importance of beta1 integrin in cardiac form and function and indicates that integrins can be linked to development of cardiomyopathies. PMID- 11884377 TI - Protease-activated receptor-2 activation causes EDHF-like coronary vasodilation: selective preservation in ischemia/reperfusion injury: involvement of lipoxygenase products, VR1 receptors, and C-fibers. AB - Activation of protease-activated receptor (PAR)-2 has been proposed to be protective in myocardial ischemia/reperfusion (I/R) injury, an effect possibly related to an action on the coronary vasculature. Therefore, we investigated the effects of PAR2 activation on coronary tone in isolated perfused rat hearts and elucidated the mechanisms of any observed effects. Although having a negligible effect on ventricular contractility, the PAR2 activating peptide SLIGRL produced an endothelium-dependent coronary vasodilatation (ED(50)=3.5 nmol). Following I/R injury, the response to SLIGRL was selectively preserved, whereas the dilator response to acetylcholine was converted to constriction. Trypsin also produced a vasodilator dose-response curve that was biphasic in nature (ED(50-1)=0.36 U, ED(50-2)=38.71 U). Desensitization of PAR2 receptors indicated that the high potency phase was mediated by PAR2. Removal of the endothelium but not treatment with L-NAME (300 micromol/L), indomethacin (5 micromol/L), or oxyhemoglobin (10 micromol/L) inhibited the response to SLIGRL and trypsin. Treatment with the K(+) channel blockers TEA (10 mmol/L), charybdotoxin (20 nmol/L)/apamin (100 nmol/L), or elevated potassium (20 mmol/L) significantly suppressed responses. Similarly, inhibition of lipoxygenase with nordihydroguaiaretic acid (1 micromol/L), eicosatetraynoic acid (1 micromol/L), or baicalein (10 micromol/L), desensitization of C-fibers using capsaicin (1 micromol/L, 20 minutes), or blockade of vanilloid (VR1) receptors using capsazepine (3 micromol/L) inhibited the responses. This study shows, for the first time, that PAR2 activation causes endothelium-dependent coronary vasodilation that is preserved after I/R injury and is not mediated by NO or prostanoids, but involves the release of an endothelium-derived hyperpolarizing factor (EDHF), possibly a lipoxygenase derived eicosanoid, and activation of VR1 receptors on sensory C-fibers. PMID- 11884378 TI - alpha(4)beta(1) Integrin activation of L-type calcium channels in vascular smooth muscle causes arteriole vasoconstriction. AB - A pathway for the regulation of vascular tone appears to involve coupling between integrins and extracellular matrix proteins or their fragments and the subsequent modulation of ion movement across the smooth muscle cell membrane. Here, we report that the activation of L-type voltage-activated Ca(2+) channels occurs through a novel interaction of alpha(4)beta(1) integrin with peptides containing the Leu-Asp-Val (LDV) integrin--binding sequence, which is found in the CS-1 region of an alternately spliced fibronectin variant. Experiments were conducted on arterioles isolated from rat skeletal muscle. Arterioles exhibited sustained concentration-dependent vasoconstriction to LDV peptides but not to Leu-Glu-Val (LEV) control peptides. The constriction was associated with increased smooth muscle cell [Ca(2+)](i), as measured by using fura 2. The response could be inhibited with a function-blocking anti--alpha(4) integrin antibody. Removal of the endothelium did not alter the vasoconstrictor response. Further experiments demonstrated that the vasoconstriction was abolished by the L-type Ca(2+) channel inhibitor nifedipine and the Src family kinase inhibitor PP2. In studies of isolated smooth muscle cells using whole-cell patch-clamp methods, the L-type current was enhanced by the LDV but not LEV peptide and was blocked by PP2 or antibodies to alpha(4) integrin. Collectively, these data indicate that activation of alpha(4)beta(1) integrin leads to enhanced influx of Ca(2+) through L-type channels by activating a tyrosine kinase pathway, leading to vasoconstriction. Involvement of integrins in the modulation of vascular tone may be particularly important in vascular responses to mechanical signals, such as pressure and flow, and to tissue injury after damage to the extracellular matrix. PMID- 11884379 TI - Retrovirally mediated overexpression of versican v3 by arterial smooth muscle cells induces tropoelastin synthesis and elastic fiber formation in vitro and in neointima after vascular injury. AB - Versican is an extracellular matrix (ECM) proteoglycan that is synthesized as multiple splice variants. In a recent study, we demonstrated that retroviral mediated overexpression of the variant V3, which lacks chondroitin sulfate (CS) chains, altered arterial smooth muscle cell (ASMC) phenotype in short-term cell culture. We now report that V3-overexpressing ASMCs exhibit significantly increased expression of tropoelastin and increased formation of elastic fibers in long-term cell cultures. In addition, V3-overexpressing ASMCs seeded into ballooned rat carotid arteries continued to overexpress V3 and, at 4 weeks after seeding, produced a highly structured neointima significantly enriched in elastic fiber lamellae. In contrast to the hydrated, myxoid neointima produced by rounded or stellate vector-alone--transduced cells, V3-expressing cells produced a compact and highly ordered neointima, which contained elongated ASMCs that were arranged in parallel arrays and separated by densely packed collagen bundles and elastic fibers. These results indicate that a variant of versican is involved in elastic fiber assembly and may represent a novel therapeutic approach to facilitate the formation of elastic fibers. PMID- 11884380 TI - Targeting CCR2 or CD18 inhibits experimental in-stent restenosis in primates: inhibitory potential depends on type of injury and leukocytes targeted. AB - A central role for leukocytes in neointimal hyperplasia after arterial injury is suspected. However, the relative importance of neutrophils and monocytes in balloon or stent-induced injury are not well understood, and mechanistic targeting of leukocyte recruitment or function is crude. We determined the temporal and spatial distribution of different leukocytes after balloon and stent induced injury in primate iliac arteries. Based on these data, we targeted neutrophil and monocyte recruitment selectively after angioplasty or stent implantation and demonstrated that monocyte-specific blockade achieved via blockade of the MCP-1 receptor CCR2, was effective at reducing neointimal hyperplasia after stenting. In contrast, combined neutrophil and monocyte blockade achieved by targeting the leukocyte beta(2)-integrin beta-subunit CD18 was required to reduce neointimal hyperplasia after balloon injury. Distinct patterns of leukocyte infiltration in balloon versus stent-injured arteries predict distinct mechanisms for antiinflammatory strategies targeting neutrophils or monocytes in primates and may assist design of effective clinical strategies for optimizing vascular interventions. PMID- 11884381 TI - Isoform-specific modulation of voltage-gated Na(+) channels by calmodulin. AB - Calmodulin (CaM) is a calcium-sensing protein that binds to Na(+) channels, with unknown functional consequences. Wild-type CaM produced a hyperpolarizing shift in the steady-state availability of expressed skeletal muscle (micro1) but not cardiac (hH1) Na(+) channels. Mutant CaM(1234) did not alter the voltage dependence or the kinetics of gating of either micro1 or hH1. Mutation of the highly conserved IQ motif in the carboxyl terminus of both isoforms (IQ/AA) slowed the kinetics of current decay and abolished the effect of wild-type CaM on micro1, but did not alter hH1 currents. The IQ/AA mutation eliminated CaM binding to the carboxyl terminus of both micro1 and hH1 channels. Inhibition of Ca(2+)/CaM kinase (CaM-K) slowed the current decay, the rate of entry into inactivation, and shifted the voltage dependence of hH1 in the depolarizing direction independent of CaM overexpression with no effect on micro1 Na(+) channels. CaM signaling modulates Na(+) currents in an isoform-specific manner, via direct interaction with skeletal muscle Na(+) channels and through CaM-K in the case of the cardiac isoform. The full text of this article is available at http://www.circresaha.org. PMID- 11884383 TI - Evidence that the transmembrane biogenesis of aquaporin 1 is cotranslational in intact mammalian cells. AB - Most polytopic membrane proteins are believed to integrate into the membrane of the endoplasmic reticulum (ER) cotranslationally. However, recent studies with Xenopus oocytes and dog pancreatic microsomes have suggested that this is not the case for human aquaporin 1 (AQP1). These experiments indicate that membrane spanning segments (MSSs) 2 and 4 of AQP1 do not integrate into the membrane cotranslationally so that this protein initially adopts a four MSS topology. A later maturation event involving a 180-degree rotation of MSS 3 from an N(lum)/C(cyt) to an N(cyt)/C(lum) orientation and the concomitant integration of MSSs 2 and 4 into the membrane results in the final six MSS topology. Here we examine the biogenesis of AQP1 in the human embryonic kidney cell line HEK-293T. To do this, we constructed an expression vector for a fusion protein consisting of the enhanced green fluorescent protein followed by an insertion site for AQP1 sequences and a C-terminal glycosylation tag. We then transiently transfected HEK 293T cells with this vector containing the AQP1 sequence truncated after each MSS. Glycosylation of the C-terminal tag was used to monitor its location relative to the ER lumen and consequently the membrane integration and orientation of successive MSSs. In contrast to previous studies our results indicate that AQP1 integrates into the ER membrane cotranslationally in intact HEK-293T cells. PMID- 11884382 TI - Effects of angiotensin II infusion on the expression and function of NAD(P)H oxidase and components of nitric oxide/cGMP signaling. AB - Angiotensin II infusion causes endothelial dysfunction by increasing NAD(P)H oxidase-mediated vascular superoxide production. However, it remains to be elucidated how in vivo angiotensin II treatment may alter the expression of the gp91(phox) isoforms and the endothelial nitric oxide synthase (NOS III) and subsequent signaling events and whether, in addition to the NAD(P)H oxidase, NOS III contributes to vascular superoxide formation. We therefore studied the influence of in vivo angiotensin II treatment (7 days) in rats on endothelial function and on the expression of the NAD(P)H oxidase subunits p22(phox), nox1, nox4, and gp91(phox) and NOS III. Further analysis included the expression of NO downstream targets, the soluble guanylyl cyclase (sGC), the cGMP-dependent protein kinase I (cGK-I), and the expression and phosphorylation of the vasodilator-stimulated phosphoprotein (VASP) at Ser239 (P-VASP). Angiotensin II caused endothelial dysfunction and increased vascular superoxide. Likewise, we found an increase in vascular protein kinase C (PKC) activity, in the expression of nox1 (6- to 7-fold), gp91(phox) (3-fold), p22(phox) (3-fold), NOS III mRNA, and protein. NOS-inhibition with N(G)-nitro-L-arginine decreased superoxide in vessels from angiotensin II-treated animals, compatible with NOS-uncoupling. Vascular NO assessed with electron paramagnetic resonance was markedly reduced. Likewise, a decrease in sGC-expression and P-VASP levels was found. In vivo PKC inhibition with chelerythrine reduced angiotensin II-induced superoxide production and markedly inhibited upregulation of NAD(P)H oxidase subunits. We therefore conclude that angiotensin II-induced increases in the activity and the expression of NAD(P)H oxidase are at least in part PKC-dependent. NADPH oxidase induced superoxide production may trigger NOS III uncoupling, leading to impaired NO/cGMP signaling and to endothelial dysfunction in this animal model. The full text of this article is available at http://www.circresaha.org. PMID- 11884384 TI - Structure of the carboxyl-terminal Src kinase, Csk. AB - The carboxyl-terminal Src kinase (Csk) is an indispensable negative regulator for the Src family tyrosine kinases (SFKs) that play pivotal roles in various cell signalings. To understand the molecular basis of the Csk-mediated regulation of SFKs, we elucidated the crystal structure of full-length Csk. The Csk crystal consists of six molecules classified as active or inactive states according to the coordinations of catalytic residues. Csk assembles the SH2 and SH3 domains differently from inactive SFKs, and their binding pockets are oriented outward enabling the intermolecular interaction. In active molecules, the SH2-kinase and SH2-SH3 linkers are tightly stuck to the N-lobe of the kinase domain to stabilize the active conformation, and there is a direct linkage between the SH2 and the kinase domains. In inactive molecules, the SH2 domains are rotated destroying the linkage to the kinase domain. Cross-correlation matrices for the active molecules reveal that the SH2 domain and the N-lobe of the kinase domain move as a unit. These observations suggest that Csk can be regulated through coupling of the SH2 and kinase domains and that Csk provides a novel built-in activation mechanism for cytoplasmic tyrosine kinases. PMID- 11884385 TI - Direct phosphorylation of capsaicin receptor VR1 by protein kinase Cepsilon and identification of two target serine residues. AB - The capsaicin receptor, VR1, is a sensory neuron-specific ion channel that serves as a polymodal detector of pain-producing chemical and physical stimuli. It has been reported that ATP, one of the inflammatory mediators, potentiates the VR1 currents evoked by capsaicin or protons and reduces the temperature threshold for activation of VR1 through metabotropic P2Y(1) receptors in a protein Kinase C (PKC)-dependent pathway, suggesting the phosphorylation of VR1 by PKC. In this study, direct phosphorylation of VR1 upon application of phorbol 12-myristate 13 acetate (PMA) was proven biochemically in cells expressing VR1. An in vitro kinase assay using glutathione S-transferase fusion proteins with cytoplasmic segments of VR1 showed that both the first intracellular loop and carboxyl terminus of VR1 were phosphorylated by PKCepsilon. Patch clamp analysis of the point mutants where Ser or Thr residues were replaced with Ala in the total 16 putative phosphorylation sites showed that two Ser residues, Ser(502) and Ser(800) were involved in the potentiation of the capsaicin-evoked currents by either PMA or ATP. In the cells expressing S502A/S800A double mutant, the temperature threshold for activation was not reduced upon PMA treatment. The two sites would be promising targets for the development of substance modulating VR1 function, thereby reducing pain. PMID- 11884386 TI - Mitogen-activated protein kinases and activator protein 1 are required for proliferation and cardiomyocyte differentiation of P19 embryonal carcinoma cells. AB - Mitogen-activated protein kinases (MAPKs) have been implicated as regulators of differentiation. The biological effect of MAPK signaling in the nucleus is achieved by signal-responsive transcription factors. Here we have investigated MAPK signaling and activation of AP-1 transcription factors in P19 embryonal carcinoma cells undergoing cardiomyocyte differentiation. We show that aggregation and Me(2)SO treatment, which trigger the differentiation response, result in sustained activation of JNK1, p38, and ERK1/2 MAPKs and acquisition of AP-1 DNA binding activity. The induced AP-1 activity consists of c-Jun, JunD, and Fra-2 proteins and is accompanied with the increased expression of these proteins. JNK is involved in c-Jun phosphorylation, whereas ERK and p38 activities are essential for maximal c-Jun and Fra-2 expression, and AP-1 DNA binding activity. While the inhibition of ERK can partially prevent the formation of beating cardiomyocytes, the activity of p38 is absolutely required for the differentiation. Expression of dominant negative c-Jun(bZIP) in P19 cells can also inhibit the differentiation response. Surprisingly, however, expression of dominant negative SEK or JNK causes an inhibition of P19 cell proliferation. Taken together, the results show that ERK, JNK, p38, and AP-1 are activated in a coordinated and sustained manner, and contribute to proliferation and cardiomyocyte differentiation of P19 cells. PMID- 11884387 TI - Biosynthesis and secretion of parathyroid hormone are sensitive to proteasome inhibitors in dispersed bovine parathyroid cells. AB - Preproparathyroid hormone (prepro-PTH) is one of the proteins abundantly synthesized by parathyroid chief cells; yet under normal growth conditions, little or no prepro-PTH can be detected in these cells. Although this may be attributed to effective cotranslational translocation and proteolytic processing, proteasome-mediated degradation of PTH precursors may be important in the regulation of the levels of these precursors and hence PTH secretion. The effects of N-acetyl-Leu-Leu-norleucinal, N-acetyl-Leu-Leu-methional, carbobenzoxy-Leu-Leu leucinal (MG132), benzyloxycarbonyl-Ile-Glu(t-butyl)-Ala-leucinal (proteasome inhibitor I), and lactacystin on the biosynthesis and secretion of PTH were examined in dispersed bovine parathyroid cells. We demonstrate that treatment of these cells with proteasome inhibitors caused the accumulation of prepro-PTH and pro-PTH. Compared with mock-treated cells, the processing of pro-PTH to PTH was delayed, and the secretion of intact PTH decreased in proteasome inhibitor treated cells. Relieving the inhibition of the proteasome by chasing MG132 treated cells in medium without the inhibitor led to the rapid disappearance of the accumulated prepro-PTH, and the rate of PTH secretion was restored to levels comparable to those in mock-treated cells. Furthermore, overexpression of the Hsp70 family of molecular chaperones was observed in proteasome inhibitor-treated cells, and we show that PTH/PTH precursors interact with these molecular chaperones. These data suggest the involvement of parathyroid cell proteasomes in the quality control of PTH biosynthesis. PMID- 11884388 TI - Characterization of the acid stability of glycosidically linked neuraminic acid: use in detecting de-N-acetyl-gangliosides in human melanoma. AB - The glycosidic linkage of sialic acids is much more sensitive to acid hydrolysis than those of other monosaccharides in vertebrates. The commonest sialic acids in nature are neuraminic acid (Neu)-based and are typically N-acylated at the C5 position. Unsubstituted Neu is thought to occur on native gangliosides of certain tumors and cell lines, and synthetic de-N-acetyl-gangliosides have potent biological properties in vitro. However, claims for their natural existence are based upon monoclonal antibodies and pulse-chase experiments, and there have been no reports of their chemical detection. Here we report that one of these antibodies shows nonspecific cross-reactivity with a polypeptide epitope, further emphasizing the need for definitive chemical proof of unsubstituted Neu on naturally occurring gangliosides. While pursuing this, we found that alpha2-3 linked Neu on chemically de-N-acetylated G(M3) ganglioside resists acid hydrolysis under conditions where the N-acetylated form is completely labile. To ascertain the generality of this finding, we investigated the stability of glycosidically linked alpha- and beta-methyl glycosides of Neu. Using NMR spectroscopy to monitor glycosidic linkage hydrolysis, we find that only 47% of Neualpha2Me is hydrolyzed after 3 h in 10 mm HCl at 80 degrees C, whereas Neu5Acalpha2Me is 95% hydrolyzed after 20 min under the same conditions. Notably, Neubeta2Me is hydrolyzed even slower than Neualpha2Me, indicating that acid resistance is a general property of glycosidically linked Neu. Taking advantage of this, we modified classical purification techniques for de-N-acetyl ganglioside isolation using acid to first eliminate conventional gangliosides. We also introduce a phospholipase-based approach to remove contaminating phospholipids that previously hindered efforts to study de-N-acetyl-gangliosides. The partially purified sample can then be N-propionylated, allowing acid release and mass spectrometric detection of any originally existing Neu as Neu5Pr. These advances allowed us to detect covalently bound Neu in lipid extracts of a human melanoma tumor, providing the first chemical proof for naturally occurring de-N acetyl-gangliosides. PMID- 11884389 TI - Inhibition of lipid raft-dependent signaling by a dystrophy-associated mutant of caveolin-3. AB - Specific point mutations in caveolin-3, a predominantly muscle-specific member of the caveolin family, have been implicated in limb-girdle muscular dystrophy and in rippling muscle disease. We examined the effect of these mutations on caveolin 3 localization and function. Using two independent assay systems, Raf activation in fibroblasts and neurite extension in PC12 cells, we show that one of the caveolin-3 point mutants, caveolin-3-C71W, specifically inhibits signaling by activated H-Ras but not by K-Ras. To gain insights into the effect of the mutant protein on H-Ras signaling, we examined the localization of the mutant proteins in fibroblastic cells and in differentiating myotubes. Unlike the previously characterized caveolin-3-DGV mutant, the inhibitory caveolin-3-C71W mutant reached the plasma membrane and colocalized with wild type caveolins. In BHK cells, caveolin-3-C71W associated with caveolae and in differentiating muscle cells with the developing T-tubule system. In contrast, the caveolin-3-P104L mutant accumulated in the Golgi complex and had no effect on H-Ras-mediated Raf activation. Inhibition by caveolin-3-C71W was rescued by cholesterol addition, suggesting that the mutant protein perturbs cholesterol-rich raft domains. Thus, we have demonstrated that a naturally occurring caveolin-3 mutation can inhibit signaling involving cholesterol-sensitive raft domains. PMID- 11884390 TI - Candoxin, a novel toxin from Bungarus candidus, is a reversible antagonist of muscle (alphabetagammadelta ) but a poorly reversible antagonist of neuronal alpha 7 nicotinic acetylcholine receptors. AB - In contrast to most short and long chain curaremimetic neurotoxins that produce virtually irreversible neuromuscular blockade in isolated nerve-muscle preparations, candoxin, a novel three-finger toxin from the Malayan krait Bungarus candidus, produced postjunctional neuromuscular blockade that was readily and completely reversible. Nanomolar concentrations of candoxin (IC(50) = approximately 10 nm) also blocked acetylcholine-evoked currents in oocyte expressed rat muscle (alphabetagammadelta) nicotinic acetylcholine receptors in a reversible manner. In contrast, it produced a poorly reversible block (IC(50) = approximately 50 nm) of rat neuronal alpha7 receptors, clearly showing diverse functional profiles for the two nicotinic receptor subsets. Interestingly, candoxin lacks the helix-like segment cyclized by the fifth disulfide bridge at the tip of the middle loop of long chain neurotoxins, reported to be critical for binding to alpha7 receptors. However, its solution NMR structure showed the presence of some functionally invariant residues involved in the interaction of both short and long chain neurotoxins to muscle (alphabetagammadelta) and long chain neurotoxins to alpha7 receptors. Candoxin is therefore a novel toxin that shares a common scaffold with long chain alpha-neurotoxins but possibly utilizes additional functional determinants that assist in recognizing neuronal alpha7 receptors. PMID- 11884391 TI - Distinct role of phosphatidylinositol 3-kinase and Rho family GTPases in Vav3 induced cell transformation, cell motility, and morphological changes. AB - Vav3 is a member of the Vav family of guanine nucleotide exchange factors (GEFs) for the Rho family GTPases. Deleting the N-terminal calponin homology (CH) domain to generate Vav3-(5-10) or deleting both the CH and the acidic domain to generate Vav3-(6-10) results in activating the transforming potential of Vav3. Expression of either the full-length Vav3 or its truncation mutants led to activation of phosphatidylinositol 3-kinase (PI3K), mitogen-activated protein kinase (MAPK), focal adhesion kinase (FAK), and Stat3. We investigated the requirement of these signaling molecules for Vav3-induced focus formation and found that PI3K and its downstream signaling molecules, Akt and p70 S6 kinase, are required, albeit to varying degrees. Inhibition of PI3K had a more dramatic effect than inhibition of MAPK on Vav3-(6-10)-induced focus formation. Activated PI3K enhanced the focus forming activity of Vav3-(6-10). Wild type FAK but not Y397F mutant FAK enhanced Vav3-(6-10)-induced focus formation. Dominant negative (dn) mutant of Stat3 resulted in a 60% inhibition of the focus-forming activity of Vav3-(6-10). Moreover, Rac1, RhoA, and to a lesser extent, Cdc42, are important for Vav3-(6 10)-induced focus formation. Constitutively activated (ca) Rac synergizes with Vav3-(6-10) in focus formation. This synergy requires signaling via Rho associated kinase (ROK) and p21-activated kinase (PAK), downstream effectors of Rac. Consistently, a ca PAK mutant enhanced, whereas a dn PAK mutant inhibited the focus-forming ability of Vav3-(6-10). Despite having potent focus-forming ability, Vav3-(6-10) has very weak colony-forming ability. This colony-forming ability of Vav3-(6-10) can be enhanced dramatically by co-expressing an activated PI3K and to some extent by co-expressing an activated PAK mutant or c-Myc. Interestingly, inhibition of PI3K and MAPK had no effect on the ability of either wild type or Vav3-(6-10) to induce cytoskeletal changes including formation of lamellipodia and filopodia in NIH 3T3 cells. Over expression of Vav3 or Vav3-(6 10) resulted in an enhancement of cell motility. This enhancement was dependent on PI3K, Rac1, and Cdc42 but not on Rho. Overall, our results show that signaling pathways of PI3K, MAPK, and Rho family GTPases are differentially required for Vav3-induced focus formation, colony formation, morphological changes, and cell motility. PMID- 11884392 TI - Crystal structure of the human estrogen sulfotransferase-PAPS complex: evidence for catalytic role of Ser137 in the sulfuryl transfer reaction. AB - Estrogen sulfotransferase (EST) transfers the sulfate group from 3' phosphoadenosine 5'-phosphosulfate (PAPS) to estrogenic steroids. Here we report the crystal structure of human EST (hEST) in the context of the V269E mutant-PAPS complex, which is the first structure containing the active sulfate donor for any sulfotransferase. Superimposing this structure with the crystal structure of hEST in complex with the donor product 3'-phosphoadenosine 5'-phosphate (PAP) and the acceptor substrate 17beta-estradiol, the ternary structure with the PAPS and estradiol molecule, is modeled. These structures have now provided a more complete view of the S(N)2-like in-line displacement reaction catalyzed by sulfotransferases. In the PAPS-bound structure, the side chain nitrogen of the catalytic Lys(47) interacts with the side chain hydroxyl of Ser(137) and not with the bridging oxygen between the 5'-phosphate and sulfate groups of the PAPS molecule as is seen in the PAP-bound structures. This conformational change of the side chain nitrogen indicates that the interaction of Lys(47) with Ser(137) may regulate PAPS hydrolysis in the absences of an acceptor substrate. Supporting the structural data, the mutations of Ser(137) to cysteine and alanine decrease gradually k(cat) for PAPS hydrolysis and transfer activity. Thus, Ser(137) appears to play an important role in regulating the side chain interaction of Lys(47) with the bridging oxygen between the 5'-phosphate and the sulfate of PAPS. PMID- 11884393 TI - Manipulation of a nuclear NAD+ salvage pathway delays aging without altering steady-state NAD+ levels. AB - Yeast deprived of nutrients exhibit a marked life span extension that requires the activity of the NAD(+)-dependent histone deacetylase, Sir2p. Here we show that increased dosage of NPT1, encoding a nicotinate phosphoribosyltransferase critical for the NAD(+) salvage pathway, increases Sir2-dependent silencing, stabilizes the rDNA locus, and extends yeast replicative life span by up to 60%. Both NPT1 and SIR2 provide resistance against heat shock, demonstrating that these genes act in a more general manner to promote cell survival. We show that Npt1 and a previously uncharacterized salvage pathway enzyme, Nma2, are both concentrated in the nucleus, indicating that a significant amount of NAD(+) is regenerated in this organelle. Additional copies of the salvage pathway genes, PNC1, NMA1, and NMA2, increase telomeric and rDNA silencing, implying that multiple steps affect the rate of the pathway. Although SIR2-dependent processes are enhanced by additional NPT1, steady-state NAD(+) levels and NAD(+)/NADH ratios remain unaltered. This finding suggests that yeast life span extension may be facilitated by an increase in the availability of NAD(+) to Sir2, although not through a simple increase in steady-state levels. We propose a model in which increased flux through the NAD(+) salvage pathway is responsible for the Sir2 dependent extension of life span. PMID- 11884394 TI - Alternative splice variants of doublecortin-like kinase are differentially expressed and have different kinase activities. AB - Alternative splicing of mRNA transcripts expands the range of protein products from a single gene locus. Several splice variants of DCLK (doublecortin-like kinase) have previously been reported. Here, we report the genomic organization underlying the splice variants of DCLK and examine the expression profile of two splice variants affecting the kinase domain of DCLK and CPG16 (candidate plasticity gene 16), one containing an Arg-rich domain and the other affecting the C terminus of the protein. These splice alternatives were differentially expressed in embryonic and adult brain. Both splice variants disrupted DCLK PEST domains; however, all splice variants remained sensitive to proteolysis by calpain. The adult-specific C-terminal splice variant of DCLK had reduced autophosphorylation activity, but similar kinase activity for myelin basic protein relative to the embryonic splice variant. The splice variant adding an Arg-rich domain gained an autophosphorylation site at Ser-382. Although this protein isoform was expressed mainly in the adult brain, the phosphorylated form was strongly enriched in embryonic brain and adult olfactory bulb, suggesting a possible role in migrating neurons. PMID- 11884395 TI - Casein kinase I and casein kinase II differentially regulate axin function in Wnt and JNK pathways. AB - Axin uses different combinations of functional domains in down-regulation of the Wnt pathway and activation of the MEKK1/JNK pathway. We are interested in the elucidation of the functional switch of Axin. In the present study, we show that the Wnt activator CKIepsilon, but not CKIIalpha, Frat1, LRP5, or LRP6, inhibited Axin-mediated JNK activation. We also found that both CKIalpha and CKIepsilon interacted with Axin, whereas CKIIalpha did not bind to Axin and had no effect on Axin-mediated JNK activity even though CKIIalpha has also been suggested to be an activator for the Wnt pathway. The COOH-terminal region and the MEKK1-interacting domain of Axin are important for CKIalpha-Axin and CKIepsilon-Axin interaction. We further demonstrated that CKIepsilon and CKIalpha binding to Axin excluded MEKK1 binding, indicating that a competitive physical occupancy may underlie the inhibitory effect. Moreover, our data indicated that CKIepsilon kinase activity plays an additive role in this effect. Taken together, we have demonstrated that CKI and CKII exhibit differential effects on Axin-MEKK1 interaction and Axin mediated JNK activation. Furthermore, our data suggest that CKI may provide a possible switch mechanism for Axin function in the regulation of Wnt and JNK pathways. PMID- 11884396 TI - Pro-inflammatory cytokines tumor necrosis factor alpha and interleukin-6 and survival factor epidermal growth factor positively regulate the murine GSTA4 enzyme in hepatocytes. AB - We hypothesized that glutathione transferases could be induced and may participate to cellular defenses against the oxidative stress occurring during liver regeneration. Here, we evidenced that murine GSTA1 (mGSTA1), A4, Pi, and Mu are up-regulated during mouse liver regeneration, exhibiting a biphasic pattern of induction correlating early G(1) phase and G(1)/S transition of the cell cycle. Using confocal microscopy immunolocalization and subcellular fractionation, mGSTA4 was demonstrated in both mitochondria and cytosol and found preferentially increased in cytosol during liver regeneration. In addition, mGSTA4 was induced in vivo and in cultured hepatocytes by tumor necrosis factor alpha (TNFalpha), interleukin-6 (IL-6), and epidermal growth factor (EGF), factors that play crucial roles in hepatocyte survival and proliferation during liver regeneration. However, the mitogenic effect of EGF was not responsible for the induction of mGSTA4. In transient transfections, IL-6 and EGF, but not TNFalpha, transactivated the human GSTA4 (hGSTA4) promoter cloned upstream of the luciferase reporter gene suggesting that IL-6 and EGF up-regulated hGSTA4 at a transcriptional level, whereas TNFalpha could rather act at a post transcriptional level. The inhibition of phosphoinositide 3-kinase, p38 MAPK, and MEK/ERK signaling pathways, using specific inhibitors, prevented EGF-dependent induction of mGSTA4 and transactivation of hGSTA4 promoter. Altogether, these data favor the conclusion that, in regenerating hepatocytes, several GST isoforms are induced and that cytokines TNFalpha and IL-6 and survival factor EGF positively regulate mGSTA4 via survival signaling pathways. PMID- 11884397 TI - Mrd1p is required for processing of pre-rRNA and for maintenance of steady-state levels of 40 S ribosomal subunits in yeast. AB - Ribosome biogenesis is a conserved process in eukaryotes that requires a large number of small nucleolar RNAs and trans-acting proteins. The Saccharomyces cerevisiae MRD1 (multiple RNA-binding domain) gene encodes a novel protein that contains five consensus RNA-binding domains. Mrd1p is essential for viability. Mrd1p partially co-localizes with the nucleolar protein Nop1p. Depletion of Mrd1p leads to a selective reduction of 18 S rRNA and 40 S ribosomal subunits. Mrd1p associates with the 35 S precursor rRNA (pre-rRNA) and U3 small nucleolar RNAs and is necessary for the initial processing at the A(0)-A(2) cleavage sites in pre-rRNA. The presence of five RNA-binding domains in Mrd1p suggests that Mrd1p may function to correctly fold pre-rRNA, a requisite for proper cleavage. Sequence comparisons suggest that Mrd1p homologues exist in all eukaryotes. PMID- 11884398 TI - Correlation between tRNALys3 aminoacylation and its incorporation into HIV-1. AB - During human immunodeficiency virus type 1 (HIV-1) assembly, tRNA(Lys) is selectively packaged into the virus, where tRNA(Lys3) serves as the primer for reverse transcription. Lysyl-tRNA synthetase is also selectively incorporated into HIV-1 and is therefore a strong candidate for being the signal by which viral proteins interact with tRNA(Lys) isoacceptors. Previously, mutations in the tRNA(Lys3) anticodon have been shown to strongly inhibit the charging of tRNA(Lys3) by lysyl-tRNA synthetase in vitro, and we show here that in vivo aminoacylation is also inhibited by anticodon changes. The order of decreasing in vivo aminoacylation for tRNA(Lys3) anticodon mutants is: wild-type SUU (where S = mcm(5)S(2)U) 100%) --> SGU (49%) --> CGU (40%) --> SGA (0%) and CGA (0%). We found that the ability of these tRNA(Lys3) anticodon variants to be aminoacylated in vivo is directly correlated with their ability to be packaged into HIV-1. These data showed that the anticodon is a major determinant for tRNA(Lys3) packaging and support the conclusion that its productive interaction with lysyl tRNA synthetase is important for tRNA(Lys3) incorporation into HIV-1. PMID- 11884399 TI - P-TEFb containing cyclin K and Cdk9 can activate transcription via RNA. AB - Different positive transcription elongation factor b (P-TEFb) complexes isolated from mammalian cells contain a common catalytic subunit (Cdk9) and the unique regulatory cyclins CycT1, CycT2a, CycT2b, or CycK. The role of CycK as a transcriptional cyclin was demonstrated in this study. First, CycK activated transcription when tethered heterologously to RNA, which required the kinase activity of Cdk9. Although this P-TEFb could phosphorylate the C-terminal domain (CTD) of RNA polymerase II (RNAPII) in vitro, in contrast to CycT1 and CycT2, CycK did not activate transcription when tethered to DNA. Interestingly, when the C termini of CycT1 and CycT2 or only the histidine-rich stretch from positions 481 to 551 in CycT1 were added to CycK, the extended chimeras activated transcription equivalently via DNA. Moreover, these transcriptional effects required the CTD of RNAPII in cells. Thus, CycK functions as P-TEFb only via RNA, which suggests the presence of cellular RNA-bound activators that require CycK for their transcriptional activity. PMID- 11884400 TI - Loss of the peroxisome proliferation-activated receptor gamma (PPARgamma ) does not affect mammary development and propensity for tumor formation but leads to reduced fertility. AB - The peroxisome proliferation-activated receptor gamma (PPARgamma) is expressed in many cell types including mammary epithelium, ovary, macrophages, and B- and T cells. PPARgamma has an anti-proliferative effect in pre-adipocytes and mammary epithelial cells, and treatment with its ligands reduced the progression of carcinogen-induced mammary tumors in mice. Because PPARgamma-null mice die in utero it has not been possible to study its role in development and tumorigenesis in vivo. To investigate whether PPARgamma is required for the establishment and physiology of different cell types, a cell-specific deletion of the gene was carried out in mice using the Cre-loxP recombination system. We deleted the PPARgamma gene in mammary epithelium using WAP-Cre transgenic mice and in epithelial cells, B- and T-cells, and ovary cells using MMTV-Cre mice. The presence of PPARgamma was not required for functional development of the mammary gland during pregnancy and for the establishment of B- and T-cells. In addition, no increase in mammary tumors was observed. However, loss of the PPARgamma gene in oocytes and granulosa cells resulted in impaired fertility. These mice have normal populations of follicles, they ovulate and develop corpora lutea. Although progesterone levels are decreased and implantation rates are reduced, the exact cause of the impaired fertility remains to be determined. PMID- 11884402 TI - Identification of ERp29, an endoplasmic reticulum lumenal protein, as a new member of the thyroglobulin folding complex. AB - Folding and post-translational modification of the thyroid hormone precursor, thyroglobulin (Tg), in the endoplasmic reticulum (ER) of the thyroid epithelial cells is facilitated by several molecular chaperones and folding enzymes, such as BiP, GRP94, calnexin, protein disulfide isomerase, ERp72, and others. They have been shown to associate simultaneously and/or sequentially with Tg in the course of its maturation, thus forming large heterocomplexes in the ER of thyrocytes. Here we present evidence that such complexes include a novel member, an ER resident lumenal protein, ERp29, which is present in all mammalian tissues with exceptionally high levels of expression in the secretory cells. ERp29 was induced upon treatment of FRTL-5 rat thyrocytes with the thyroid-stimulating hormone, which is essential for the maintenance of thyroid cells and Tg biosynthesis. Chemical cross-linking followed by the cell lysis and immunoprecipitation of ERp29 or Tg revealed association of these proteins and additionally, immunocomplexes that also included major ER chaperones, BiP and GRP94. Sucrose density gradient analysis indicated co-localization of ERp29 with Tg and BiP in the fractions containing large macromolecular complexes. This was supported by immunofluorescent microscopy showing co-localization of ERp29 with Tg in the putative transport vesicular structures. Affinity chromatography using Tg as an affinity ligand demonstrated that ERp29 might be selectively isolated from the FRTL-5 cell lysate or purified lumenal fraction of rat liver microsomes along with the other ER chaperones. Preferential association with the urea-denatured Tg Sepharose was indicative of either direct or circuitous ERp29/Tg interactions in a chaperone-like manner. Despite the presence of the C-terminal ER-retrieval signal, significant amounts of ERp29 were also recovered from the culture medium of stimulated thyrocytes, indicating ERp29 secretion. Based on these data, we suggest that the function of ERp29 in thyroid cells is connected with folding and/or secretion of Tg. PMID- 11884401 TI - Protein kinase C and ERK activation are required for TFF-peptide-stimulated bronchial epithelial cell migration and tumor necrosis factor-alpha-induced interleukin-6 (IL-6) and IL-8 secretion. AB - TFF-peptides (formerly P-domain peptides, trefoil factors) are typical secretory products of many mucous epithelia and are aberrantly secreted during chronic inflammatory diseases. They are known to enhance the migration of intestinal, corneal, and bronchial epithelial cells. Using the human bronchial epithelial cell line BEAS-2B as a model, it is shown here for the first time that TFF peptides are capable of modulating the inflammatory response in vitro by regulating tumor necrosis factor-alpha-induced secretion of interleukin (IL)-6 and IL-8. In contrast, TFF2 itself does not change IL-6 and IL-8 secretion but triggers sustained activation of the extracellular signal-regulated kinases (ERK1/2) as well as phosphorylation of c-Jun N-terminal kinase (JNK). A complex differential regulation of tumor necrosis factor-alpha-induced IL-6 and IL-8 secretion by TFF2 is observed that involves signaling via protein kinase C and ERK1/2. Furthermore, the motogenic effect of TFF2 on BEAS-2B cells is analyzed using a modified Boyden chamber assay. This migratory effect is shown to be dependent not only on protein kinase C and ERK1/2 but also on the activation of the Src family of tyrosine kinases. Taken together, the data presented indicate an important physiological role of TFF-peptides during inflammatory conditions of mucous epithelia. PMID- 11884403 TI - Shedding of the interleukin-6 (IL-6) receptor (gp80) determines the ability of IL 6 to induce gp130 phosphorylation in human osteoblasts. AB - Human osteoblasts produce interleukin-6 (IL-6) and respond to IL-6 in the presence of soluble IL-6 receptor (sIL-6R), but the cell surface expression of IL 6R and the mechanism of sIL-6R production are largely unknown. Three different human osteoblast-like cell lines (MG-63, HOS, and SaOS-2) and bone marrow-derived primary human osteoblasts expressed both IL-6R and gp130 as determined by flow cytometry and immunoprecipitation. However, the membrane-bound IL-6R was nonfunctional, as significant tyrosine phosphorylation of gp130 did not occur in the presence of IL-6. Phorbol myristate acetate induced a dramatic increase of both IL-6R shedding (i.e. the production of sIL-6R) and IL-6 release in osteoblast cultures, but the cell surface expression of gp130 remained unchanged. IL-6 complexed with sIL-6R, either exogenously introduced or derived from the nonfunctional cell surface form by shedding, induced rapid tyrosine phosphorylation of gp130. This effect was inhibited by neutralizing antibodies to either sIL-6R or gp130, indicating that the gp130 activation was induced by IL 6/sIL-6R/gp130 interaction. Protein kinase C inhibitors blocked phorbol myristate acetate-induced and spontaneous shedding of IL-6R resulting in the absence of sIL 6R in the culture medium, which in turn also prevented the activation of gp130. In conclusion, human osteoblasts express cell surface IL-6R, which is unable to transmit IL-6-induced signals until it is shed into its soluble form. This unique mechanism provides the flexibility for osteoblasts to control their own responsiveness to IL-6 via the activation of an IL-6R sheddase, resulting in an immediate production of functionally active osteoblast-derived sIL-6R. PMID- 11884405 TI - Investigation of the roles of catalytic residues in serotonin N acetyltransferase. AB - Serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase (AANAT)) is a critical enzyme in the light-mediated regulation of melatonin production and circadian rhythm. It is a member of the GNAT (GCN-5-related N-acetyltransferase) superfamily of enzymes, which catalyze a diverse array of biologically important acetyl transfer reactions from antibiotic resistance to chromatin remodeling. In this study, we probed the functional properties of two histidines (His-120 and His-122) and a tyrosine (Tyr-168) postulated to be important in the mechanism of AANAT based on prior x-ray structural and biochemical studies. Using a combination of steady-state kinetic measurements of microviscosity effects and pH dependence on the H122Q, H120Q, and H120Q/H122Q AANAT mutants, we show that His 122 (with an apparent pK(a) of 7.3) contributes approximately 6-fold to the acetyltransferase chemical step as either a remote catalytic base or hydrogen bond donor. Furthermore, His-120 and His-122 appear to contribute redundantly to this function. By analysis of the Y168F AANAT mutant, it was demonstrated that Tyr-168 contributes approximately 150-fold to the acetyltransferase chemical step and is responsible for the basic limb of the pH-rate profile with an apparent (subnormal) pK(a) of 8.5. Paradoxically, Y168F AANAT showed 10-fold enhanced apparent affinity for acetyl-CoA despite the loss of a hydrogen bond between the Tyr phenol and the CoA sulfur atom. The X-ray crystal structure of Y168F AANAT bound to a bisubstrate analog inhibitor showed no significant structural perturbation of the enzyme compared with the wild-type complex, but revealed the loss of dual inhibitor conformations present in the wild-type complex. Taken together with kinetic measurements, these crystallographic studies allow us to propose the relevant structural conformations related to the distinct alkyltransferase and acetyltransferase reactions catalyzed by AANAT. These findings have significant implications for understanding GNAT catalysis and the design of potent and selective inhibitors. PMID- 11884404 TI - Overexpression and ribozyme-mediated targeting of transcriptional coactivators CREB-binding protein and p300 revealed their indispensable roles in adipocyte differentiation through the regulation of peroxisome proliferator-activated receptor gamma. AB - The cAMP-response element-binding protein-binding protein (CBP) and p300 are common coactivators for several transcriptional factors. It has been reported that both CBP and p300 are significant for the activation of peroxisome proliferator-activated receptor gamma (PPARgamma), which is a crucial nuclear receptor in adipogenesis. However, it remains unclear whether CBP and/or p300 is physiologically essential to the activation of PPARgamma in adipocytes and adipocyte differentiation. In this study, we investigated the physiological significance of CBP/p300 in NIH3T3 cells transiently expressing PPARgamma and CBP and in 3T3-L1 preadipocytes stably expressing CBP- or p300-specific ribozymes. In PPARgamma-transfected NIH3T3 cells, induction of expression of PPARgamma target genes such as adipocyte fatty acid-binding protein (aP2) and lipoprotein lipase (LPL) by adding thiazolidinedione was enhanced, depending on the amount of a CBP expression plasmid transfected. Expression of aP2 and LPL genes, as well as glycerol-3-phosphate dehydrogenase activity and triacylglyceride accumulation after adipogenic induction, was largely suppressed in 3T3-L1 adipocytes expressing either the CBP- or p300-specific active ribozyme, but not in inactive ribozyme-expressing cells. These data suggest that both CBP and p300 are indispensable for the full activation of PPARgamma and adipocyte differentiation and that CBP and p300 do not mutually complement in the process. PMID- 11884406 TI - Interactions between the isolated oxygenase and reductase domains of neuronal nitric-oxide synthase: assessing the role of calmodulin. AB - Nitric-oxide synthase (NOS) is a fusion protein composed of an oxygenase domain with a heme-active site and a reductase domain with an NADPH binding site and requires Ca(2+)/calmodulin (CaM) for NO formation activity. We studied NO formation activity in reconstituted systems consisting of the isolated oxygenase and reductase domains of neuronal NOS with and without the CaM binding site. Reductase domains with 33-amino acid C-terminal truncations were also examined. These were shown to have faster cytochrome c reduction rates in the absence of CaM. N(G)-hydroxy-l-Arg, an intermediate in the physiological NO synthesis reaction, was found to be a viable substrate. Turnover rates for N(G)-hydroxy-l Arg in the absence of Ca(2+)/CaM in most of the reconstituted systems were 2.3 3.1 min(-1). Surprisingly, the NO formation activities with CaM binding sites on either reductase or oxygenase domains were decreased dramatically on addition of Ca(2+)/CaM. However, NADPH oxidation and cytochrome c reduction rates were increased by the same procedure. Activation of the reductase domains by CaM addition or by C-terminal deletion failed to increase the rate of NO synthesis. Therefore, both mechanisms appear to be less important than the domain-domain interaction, which is controlled by CaM binding in wild-type neuronal NOS, but not in the reconstituted systems. PMID- 11884407 TI - Molecular and biochemical characterization of a highly stable bacterial laccase that occurs as a structural component of the Bacillus subtilis endospore coat. AB - The Bacillus subtilis endospore coat protein CotA shows laccase activity. By using comparative modeling techniques, we were able to derive a model for CotA based on the known x-ray structures of zucchini ascorbate oxidase and Cuprinus cereneus laccase. This model of CotA contains all the structural features of a laccase, including the reactive surface-exposed copper center (T1) and two buried copper centers (T2 and T3). Single amino acid substitutions in the CotA T1 copper center (H497A, or M502L) did not prevent assembly of the mutant proteins into the coat and did not alter the pattern of extractable coat polypeptides. However, in contrast to a wild type strain, both mutants produced unpigmented colonies and spores unable to oxidize syringaldazine (SGZ) and 2'2-azino-bis-(3 ethylbenzothiazoline-6-sulfonic acid) (ABTS). The CotA protein was purified to homogeneity from an overproducing Escherichia coli strain. The purified CotA shows an absorbance and a EPR spectra typical of blue multicopper oxidases. Optimal enzymatic activity was found at < or =pH 3.0 and at pH 7.0 for ABTS or SGZ oxidation, respectively. The apparent K(m) values for ABTS and SGZ at 37 degrees C were of 106 +/- 11 and 26 +/- 2 microm, respectively, with corresponding k(cat) values of 16.8 +/- 0.8 and 3.7 +/- 0.1 s(-1). Maximal enzyme activity was observed at 75 degrees C with ABTS as substrate. Remarkably, the coat-associated or the purified enzyme showed a half-life of inactivation at 80 degrees C of about 4 and 2 h, respectively, indicating that CotA is intrinsically highly thermostable. PMID- 11884408 TI - Oncogenic H-Ras enhances DNA repair through the Ras/phosphatidylinositol 3 kinase/Rac1 pathway in NIH3T3 cells. Evidence for association with reactive oxygen species. AB - This study investigated the role of oncogenic H-Ras in DNA repair capacity in NIH3T3 cells. Expression of dominant-positive H-Ras (V12-H-Ras) enhanced the host cell reactivation of luciferase activity from UV-irradiated and cisplatin-treated plasmids and also increased the unscheduled DNA synthesis following cisplatin or UV treatment of cells. This observed enhancement of DNA repair capacity was inhibited by transient transfection with dominant-negative H-Ras (N17-H-Ras) or Rac1 (N17-Rac1) plasmids. Moreover, stable transfection of dominant-positive Rac1 (V12-Rac1) further enhanced DNA repair capacity. Because reactive oxygen species (ROS) are known to be a downstream effector of oncogenic Ras, we examined the role of ROS in DNA repair capacity. We found that ROS production by V12-H-Ras expression was mediated by the Ras/phosphatidylinositol 3-kinase (PI3K)/Rac1/NADPH oxidase-dependent pathway and that pretreatment of V12-H-Ras transformed cells with an antioxidant (N-acetylcysteine) and an NADPH oxidase inhibitor (diphenyleneiodonium) decreased DNA repair capacity. Similarly, treatment with PI3K inhibitors (wortmannin and LY294002) inhibited the ability of oncogenic H-Ras to enhance DNA repair capacity. Furthermore, inhibition of the Ras/PI3K/Rac1/NADPH oxidase pathway resulted in increased sensitivity to cisplatin and UV in V12-H-Ras-expressing NIH3T3 cells. Taken together, these results provide evidence that oncogenic H-Ras activates DNA repair capacity through the Ras/PI3K/Rac1/NADPH oxidase-dependent pathway and that increased ROS production via this signaling pathway is required for enhancement of the DNA repair capacity induced by oncogenic H-Ras. PMID- 11884409 TI - S phase and meristem-specific expression of the tobacco RNR1b gene is mediated by an E2F element located in the 5' leader sequence. AB - The RB/E2F pathway is involved in the control of the G(1)/S transition of the eukaryotic cell cycle where various S phase genes are activated by specific E2F factors. Ribonucleotide reductase (RNR) plays an essential role in the DNA synthesis pathway. Earlier studies showed that there are at least two RNR1 genes (RNR1a and RNR1b) and one RNR2 gene in tobacco. In synchronized tobacco BY2 cells, RNR1b gene expression is at its highest level in S phase. To investigate transcriptional regulation of the RNR1b gene, its promoter region was cloned and sequenced. Unlike its animal counterparts, the tobacco RNR1b promoter contains a consensus E2F-binding site. Surprisingly, this site is found in the leader sequence of the gene. We show here by gel shift analysis and antibody competition that one nuclear complex specifically binds this motif, and an E2F factor is part of this complex. Using reporter gene analysis, tobacco RNR1b promoter activity was detected during S phase in synchronized cells and in plant meristematic tissues. Mutation of the E2F element substantially reduced both activities. For the first time in plants, a single E2F motif found in the leader sequence plays an important role in the meristem and S phase-specific expression of the tobacco RNR1b gene. PMID- 11884410 TI - Role of phosphatidylserine exposure and sugar chain desialylation at the surface of influenza virus-infected cells in efficient phagocytosis by macrophages. AB - HeLa cells infected with influenza A virus undergo typical caspase-dependent apoptosis and are efficiently phagocytosed by mouse peritoneal macrophages in a manner mediated by the membrane phospholipid phosphatidylserine, which is translocated to the surface of virus-infected cells during apoptosis. However, the extent of phagocytosis is not always parallel with the level of phosphatidylserine externalization. Here we examined the involvement of influenza virus neuraminidase (NA) in efficient phagocytosis of virus-infected cells. HeLa cells infected with an influenza virus strain expressing temperature-sensitive NA underwent apoptosis and produced viral proteins, including the defective NA, at a non-permissive temperature to almost the same extent as cells infected with the wild-type virus. The cells were, however, phagocytosed by macrophages with reduced efficiency. In addition, phagocytosis of cells infected with the wild type virus was severely inhibited when the cells had been maintained in the presence of the NA inhibitor zanamivir. On the other hand, the binding of sialic acid-recognizing lectins to the cell surface declined after infection with the wild-type virus. The decrease in the extent of lectin binding was greatly attenuated when cells were infected with the mutant virus or when wild-type virus infected cells were maintained in the presence of zanamivir. These results indicate that sugar chains are desialylated by NA at the surface of virus infected cells. We conclude that the presence of both phosphatidylserine and asialoglycomoieties on the cell surface is required for efficient phagocytosis of influenza virus-infected cells by macrophages. PMID- 11884411 TI - Discoidin domain receptor 2 interacts with Src and Shc following its activation by type I collagen. AB - Discoidin domain receptor 2 (DDR2) is an unusual receptor tyrosine kinase in that its ligand is fibrillar collagen rather than a growth factor-like peptide. We examined signal transduction pathways of DDR2. Here we show that DDR2 is also unusual in that it requires Src activity to be maximally tyrosine-phosphorylated, and that Src activity also promotes association of DDR2 with Shc. The interaction with Shc involves a portion of Shc not previously implicated in interaction with receptor tyrosine kinases. These results identify Src kinase and the adaptor protein Shc as key signaling intermediates in DDR2 signal transduction. Furthermore, Src is required for DDR2-mediated transactivation of the matrix metalloproteinase-2 promoter. The data support a model in which Src and the DDR2 receptor cooperate in a regulated fashion to direct the phosphorylation of both the receptor and its targets. PMID- 11884412 TI - Control of Ser2448 phosphorylation in the mammalian target of rapamycin by insulin and skeletal muscle load. AB - We have investigated the effects of insulin, amino acids, and the degree of muscle loading on the phosphorylation of Ser(2448), a site in the mammalian target of rapamycin (mTOR) phosphorylated by protein kinase B (PKB) in vitro. Phosphorylation was assessed by immunoblotting with a phosphospecific antibody (anti-Ser(P)(2448)) and with mTAb1, an activating antibody whose binding is inhibited by phosphorylation in the region of mTOR that contains Ser(2448). Incubating rat diaphragm muscles with insulin increased Ser(2448) phosphorylation but did not change the total amount of mTOR. Insulin, but not amino acids, activated PKB, as evidenced by increased phosphorylation of both Ser(308) and Thr(473) in the kinase. Ser(2448) phosphorylation was also modulated by muscle loading. Overloading the rat plantaris muscle by synergist muscle ablation, which promotes hypertrophy of the plantaris muscle, increased Ser(2448) phosphorylation. In contrast, unloading the gastrocnemius muscle by hindlimb suspension, which promotes atrophy of the muscle, decreased Ser(2448) phosphorylation, an effect that was fully reversible. Neither overloading nor hindlimb suspension significantly changed the total amount of mTOR. In summary, our results demonstrate that atrophy and hypertrophy of skeletal muscle are associated with decreases and increases in Ser(2448) phosphorylation, suggesting that modulation of this site may have an important role in the control of protein synthesis. PMID- 11884413 TI - Vitamin C prevents DNA mutation induced by oxidative stress. AB - The precise role of vitamin C in the prevention of DNA mutations is controversial. Although ascorbic acid has strong antioxidant properties, it also has pro-oxidant effects in the presence of free transition metals. Vitamin C was recently reported to induce the decomposition of lipid hydroperoxides independent of metal interactions, suggesting that it may cause DNA damage. To directly address the role of vitamin C in maintaining genomic integrity we developed a genetic system for quantifying guanine base mutations induced in human cells under oxidative stress. The assay utilized a plasmid construct encoding the cDNA for chloramphenicol acetyl transferase modified to contain an amber stop codon, which was restored to wild type by G to T transversion induced by oxidative stress. The mutation frequency was determined from the number of plasmids containing the wild type chloramphenicol acetyl transferase gene rescued from oxidatively stressed cells. Cells were loaded with vitamin C by exposing them to dehydroascorbic acid, thereby avoiding transition metal-related pro-oxidant effects of ascorbic acid. We found that vitamin C loading resulted in substantially decreased mutations induced by H(2)O(2). Depletion of glutathione led to cytotoxicity and an increase in H(2)O(2)-induced mutation frequency; however, mutation frequency was prominently decreased in depleted cells preloaded with vitamin C. The mutation results correlated with a decrease in total 8-oxo guanine measured in genomic DNA of cells loaded with vitamin C and oxidatively stressed. These findings directly support the concept that high intracellular concentrations of vitamin C can prevent oxidation-induced mutations in human cells. PMID- 11884414 TI - BH-3-only BIK functions at the endoplasmic reticulum to stimulate cytochrome c release from mitochondria. AB - Stimulation of apoptosis by p53 is accompanied by induction of the BH-3-only proapoptotic member of the BCL-2 family, BIK, and ectopic expression of BIK in p53-null cells caused the release of cytochrome c from mitochondria and activation of caspases, dependent on a functional BH-3 domain. A significant fraction of BIK, which contains a predicted transmembrane segment at its COOH terminus, was found inserted in the endoplasmic reticulum (ER) membrane, with the bulk of the protein facing the cytosol. Restriction of BIK to this membrane by replacing its transmembrane segment with the ER-selective membrane anchor of cytochrome b(5) also retained the cytochrome c release and cell death-inducing activity of BIK. Whereas induction of cell death by BIK was strongly inhibited by the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, the inhibitor was without effect on the ability of BIK to stimulate egress of cytochrome c from mitochondria. This benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone-insensitive pathway for stimulating cytochrome c release from mitochondria by ER BIK was successfully reconstituted in vitro and identified the requirement for components present in the light membrane (ER) and cytosol as necessary for this activity. Collectively, the results identify BIK as an initiator of cytochrome c release from mitochondria operating from a location at the ER. PMID- 11884415 TI - Association of tapasin and COPI provides a mechanism for the retrograde transport of major histocompatibility complex (MHC) class I molecules from the Golgi complex to the endoplasmic reticulum. AB - Tapasin is a subunit of the transporter associated with antigen processing (TAP). It associates with the major histocompatibility complex (MHC) class I. We show that tapasin interacts with beta- and gamma-subunits of COPI coatomer. COPI retrieves membrane proteins from the Golgi network back to the endoplasmic reticulum (ER). The COPI subunit-associated tapasin also interacts with MHC class I molecules suggesting that tapasin acts as the cargo receptor for packing MHC class I molecules as cargo proteins into COPI-coated vesicles. In tapasin mutant cells, neither TAP nor MHC class I are detected in association with the COPI coatomer. Interestingly, tapasin-associated MHC class I molecules are antigenic peptide-receptive and detected in both the ER and the Golgi. Our data suggest that tapasin is required for the COPI vesicle-mediated retrograde transport of immature MHC class I molecules from the Golgi network to the ER. PMID- 11884417 TI - Alternate translation occurs within the core coding region of the hepatitis C viral genome. AB - The majority of hepatitis C virus (HCV) isolates contain an open reading frame (ORF) overlapping with the core coding sequences in the +1 frame, which was assumed to be untranslated. We present evidence supporting the expression of this ORF (designated core+1 ORF) via novel translation mechanisms. First, fusion of the luciferase gene with the HCV-1 core+1 ORF followed by in vitro translation resulted in the synthesis of a chimeric protein (core+1-luciferase) that exhibited approximately 54% luciferase activity relative to the positive control (core-luciferase). Second, antisera raised against two different synthetic core+1 peptides recognized the previously identified p16 (but not p21) core protein band expressed from HCV-1, indicating the presence of epitopes from the core+1 ORF within the p16 protein. Third, HCV-positive sera specifically recognized lysates of Escherichia coli cells expressing recombinant core+1 protein, suggesting the presence of anti-core+1 antibodies in HCV-infected patients. Finally, luciferase tagging experiments designed to assess for -1 frameshifting combined with site directed mutagenesis experiments supported the presence of +1/-1 ribosomal frameshift translation mechanisms within the core coding region. In conclusion, our data provide evidence for novel translation mechanisms within the core coding region and demonstrate the expression of the core+1 ORF, at least for some HCV isolates. PMID- 11884416 TI - Processing of pro-atrial natriuretic peptide by corin in cardiac myocytes. AB - Corin is a type II transmembrane serine protease abundantly expressed in the heart. In a previous study using transfected 293 cells, we showed that corin converted pro-atrial natriuretic peptide (pro-ANP) to atrial natriuretic peptide (ANP), suggesting that corin is likely the pro-ANP convertase. Because other serine proteases such as thrombin and kallikrein had previously also been shown to cleave pro-ANP in vitro, it remained to demonstrate that corin is indeed the endogenous pro-ANP convertase in cardiomyocytes. In this study, we examined pro ANP processing in a murine cardiac muscle cell line, HL-5. Northern analysis showed that corin mRNA was present in HL-5 cells. In HL-5 cells transfected with a plasmid expressing pro-ANP, recombinant pro-ANP was converted to mature ANP as determined by Western analysis, indicating the presence of the endogenous pro-ANP convertase in these cells. The processed recombinant ANP was shown to be active in an enzyme-linked immunosorbent assay-based cGMP assay in baby hamster kidney cells. The processing of recombinant pro-ANP in HL-5 cells was highly sequence specific, because mutation R98A, but not mutations R101A and R102A, in pro-ANP prevented the conversion of pro-ANP to ANP. Expression of recombinant wild-type corin enhanced the processing of pro-ANP in HL-5 cells. In contrast, overexpression of active site mutant corin S985A or transfection of oligonucleotide small interfering RNA duplexes directed against the mouse corin gene completely inhibited the processing of recombinant pro-ANP in HL-5 cells. These results indicate that corin is the physiological pro-ANP convertase in cardiac myocytes. PMID- 11884418 TI - Mice deficient in the insulin-regulated membrane aminopeptidase show substantial decreases in glucose transporter GLUT4 levels but maintain normal glucose homeostasis. AB - The insulin-regulated aminopeptidase (IRAP) is a zinc-dependent membrane aminopeptidase. It is the homologue of the human placental leucine aminopeptidase. In fat and muscle cells, IRAP colocalizes with the insulin responsive glucose transporter GLUT4 in intracellular vesicles and redistributes to the cell surface in response to insulin, as GLUT4 does. To address the question of the physiological function of IRAP, we generated mice with a targeted disruption of the IRAP gene (IRAP-/-). Herein, we describe the characterization of these mice with regard to glucose homeostasis and regulation of GLUT4. Fed and fasted blood glucose and insulin levels in the IRAP-/- mice were normal. Whereas IRAP-/- mice responded to glucose administration like control mice, they exhibited an impaired response to insulin. Basal and insulin-stimulated glucose uptake in extensor digitorum longus muscle, and adipocytes isolated from IRAP-/- mice were decreased by 30-60% but were normal for soleus muscle from male IRAP-/- mice. Total GLUT4 levels were diminished by 40-85% in the IRAP-/- mice in the different muscles and in adipocytes. The relative distribution of GLUT4 in subcellular fractions of basal and insulin-stimulated IRAP-/- adipocytes was the same as in control cells. We conclude that IRAP-/- mice maintain normal glucose homeostasis despite decreased glucose uptake into muscle and fat cells. The absence of IRAP does not affect the subcellular distribution of GLUT4 in adipocytes. However, it leads to substantial decreases in GLUT4 expression. PMID- 11884420 TI - Cutting edge: identification of c-Rel-dependent and -independent pathways of IL 12 production during infectious and inflammatory stimuli. AB - The production of IL-12 is required for immunity to many intracellular pathogens. Recent studies have shown that c-Rel, a member of the NF-kappaB family of transcription factors, is essential for LPS-induced IL-12p40 production by macrophages. In this study, we demonstrate that c-Rel is also required for IL 12p40 production by macrophages in response to Corynebacterium parvum, CpG oligodeoxynucleotides, anti-CD40 and low molecular weight hyaluronic acid. However, c-Rel(-/-) mice infected with Toxoplasma gondii produce comparable amounts of IL-12p40 to infected wild-type mice and have an IL-12-dependent mechanism of resistance to this infection. Furthermore, c-Rel was not required for IL-12p40 production by macrophages or dendritic cells in response to soluble Toxoplasma Ag, and neutrophils from c-Rel(-/-) mice contain normal amounts of preformed IL-12p40. Together these studies reveal the presence of c-Rel-dependent pathways critical for IL-12p40 production in response to inflammatory stimuli and demonstrate a novel c-Rel-independent pathway of IL-12p40 production during toxoplasmosis. PMID- 11884419 TI - Cutting edge: inhibitory functions of the killer cell lectin-like receptor G1 molecule during the activation of mouse NK cells. AB - The killer cell lectin-like receptor G1 (KLRG1) is the mouse homolog of the rat mast cell function-associated Ag and contains an immunoreceptor tyrosine-based inhibitory motif in its cytoplasmic domain. In this study we demonstrate that both pathogenic and nonpathogenic in vivo activation of NK cells induces the expression of KLRG1 on their cell surface. Upon infection with murine CMV, this induction peaks between days 5 and 7 with about 90% of the NK cells expressing KLRG1. On day 1.5 post-murine CMV infection of C57BL/6 mice, the main producers of IFN-gamma are the KLRG1-negative NK cells. This effect has been recapitulated in vitro as we show that engagement of KLRG1 on a transfected NK cell line inhibits both cytokine production and NK cell-mediated cytotoxicity. Taken together, these data illustrate the crucial role played by KLRG1 during the termination of mouse NK cell activation. PMID- 11884421 TI - Cutting edge: recruitment of the ancestral fyn gene during emergence of the adaptive immune system. AB - The adaptive immune system (AIS) is characterized by the MHC molecules and the rearranging Ag receptors, and was established in a common ancestor of jawed vertebrates. Fyn, a Src-family tyrosine kinases, is important for normal development and function of T lymphocytes and neuronal cells. Indeed, as the result of an alternative splicing of a distinct exon 7, fyn encodes for two isoforms, FynT in T lymphocytes and FynB in the brain. How this alternative splicing of fyn transcripts has emerged and evolved in relation to the setting of the AIS remains to be established. In this study, we show that exon capture in a vertebrate ancestor by the fynT-like gene has yielded a novel fyn-encoded isoform, fynB. Unexpectedly, the newly established AIS recruited the ancestral Fyn isoform, FynT, whereas the CNS expresses the most recent one, FynB. These results shed new light on the emergence of the AIS. PMID- 11884422 TI - Cutting edge: CD83 regulates the development of cellular immunity. AB - We recently found that human CD83, a marker of mature dendritic cells, is an adhesion receptor that binds to resting monocytes and a subset of activated CD8(+) T cells. We injected CD83-Ig into mice transplanted with the immunogenic P815 mastocytoma and showed that it significantly enhanced the rate of tumor growth and inhibited the development of cytotoxic T cells. In contrast, mice immunized with CD83-transfected K1735 cells, a poorly immunogenic melanoma, could prevent the outgrowth of wild-type K1735 cells. Studies performed in vitro with human PBL showed that coimmobilized CD83-Ig and anti-CD3 enhanced T cell proliferation and increased the proportion of CD8(+) T cells. CD83-transfected B lymphoblastoid T51 cells stimulated T cell proliferation more effectively than untransfected T51 cells in MLR cultures and increased the generation of cytolytic T cells. We conclude that CD83 is a functionally important receptor that can regulate the development of cellular immunity by interacting with its ligand(s). PMID- 11884423 TI - Cutting edge: VacA, a vacuolating cytotoxin of Helicobacter pylori, directly activates mast cells for migration and production of proinflammatory cytokines. AB - Mucosal mast cells strategically located at the optimal site interact with invading bacteria. Presence of VacA, the virulent Helicobacter pylori cytotoxin, is correlated with the severity of H. pylori-induced gastritis. To examine the mechanisms of inflammation in H. pylori-induced gastritis, we administered VacA to the mice. Inoculation of VacA resulted in epithelium vacuolization and marked infiltrations of mast cells and mononuclear cells into the mucosal epithelium within 24 h. In an in vitro study using bone marrow-derived mast cells, VacA directly bound and showed a chemotactic activity to the mast cell. In addition, VacA induced bone marrow-derived mast cells to produce proinflammatory cytokines, TNF-alpha, macrophage-inflammatory protein-1alpha, IL-1beta, IL-6, IL-10, and IL 13 in a dose-dependent manner without causing degranulation. The present study suggests that early activation of mast cells by VacA may be the host early response to clear the bacteria and also may contribute to the pathogenesis of H. pylori-induced gastritis. PMID- 11884424 TI - Stromal-derived factor 1 expression in the human thymus. AB - Stromal-derived factor-1 (SDF-1), the only known ligand for the chemokine receptor CXCR4, is broadly expressed in cells of both the immune and central nervous systems, and it can induce the migration of resting leukocytes and hemopoietic progenitors. SDF-1 mRNA was previously detected in human thymus derived stromal cells, but its role in thymopoiesis was unknown. Here we show that SDF-1 is expressed in medullar epithelial cells forming Hassall's corpuscles (HC). In search of the cell type that may be attracted by SDF-1(+) cells in the medulla, we determined that dendritic cells (DC) could be found in situ in close proximity to SDF-1(+) epithelial cells in HC. In HIV-1-infected SCID-hu thymuses, DC contained apoptotic cells and were located within enlarged HC. It was further demonstrated that uptake of apoptotic thymocytes by immature DC induced an increase in CXCR4 expression and SDF-1-mediated chemotaxis. Our data suggest a role for SDF-1 in the elimination of apoptotic thymocytes. PMID- 11884425 TI - A role for cathepsin L and cathepsin S in peptide generation for MHC class II presentation. AB - The enzymes that degrade proteins to peptides for presentation on MHC class II molecules are poorly understood. The cysteinal lysosomal proteases, cathepsin L (CL) and cathepsin S (CS), have been shown to process invariant chain, thereby facilitating MHC class II maturation. However, their role in Ag processing is not established. To examine this issue, we generated embryonic fibroblast lines that express CL, CS, or neither. Expression of CL or CS mediates efficient degradation of invariant chain as expected. Ag presentation was evaluated using T cell hybridoma assays as well as mass spectroscopic analysis of peptides eluted from MHC class II molecules. Interestingly, we found that the majority of peptides are presented regardless of CL or CS expression, although these proteases often alter the relative levels of the peptides. However, for a subset of Ags, epitope generation is critically regulated by CL or CS. This result suggests that these cysteinal proteases participate in Ag processing and generate qualitative and quantitative differences in the peptide repertoires displayed by MHC class II molecules. PMID- 11884426 TI - Homeostatic regulation of intestinal villous epithelia by B lymphocytes. AB - The epithelial cell of the small intestine is one of the most rapidly regenerating cells in the body. However, the cellular mechanism and biological significance underlying this rapid regeneration remain elusive. In this study we examined the intestinal epithelia of mutant mice that lack B and/or T cells and those of normal littermates. The absence of B cells in Ig mu-chain mutant mice or B and T cells in recombination-activating gene (RAG)-2(-/-) as well as SCID mutant mice was associated with a marked acceleration of epithelial cell turnover and an up-regulation of the expression of MHC class II molecules. No such effects were observed in T cell-deficient TCR-delta and -beta double-mutant mice. As far as the goblet cells of villous epithelium are concerned, absolute numbers of them remained the same among these mutant mice that have no B and/or T cells. Alymphoplasia (aly/aly) mutant mice that lacked Peyer's patches and Ig-producing cells in the lamina propria, but harbored a large number of intestinal mucosal T cells, also displayed a significant acceleration of epithelial cell turnover and, to some extent, up-regulated expression of MHC class II molecules. Notably, the accelerated epithelial cell turnover was not observed and returned to normalcy in the Ig mu-chain mutant mice that had been given antibiotic-containing water. These findings indicate that B cells down-regulate the generation and differentiation of intestinal epithelial cells in the normal wild-type condition and suggest that enteric microorganisms are implicated in the accelerated generation of epithelial cells in mice that have no B cells. PMID- 11884427 TI - DC-SIGN (CD209) expression is IL-4 dependent and is negatively regulated by IFN, TGF-beta, and anti-inflammatory agents. AB - Dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) is a monocyte derived dendritic cell (MDDC)-specific lectin which participates in dendritic cell (DC) migration and DC-T lymphocyte interactions at the initiation of immune responses and enhances trans-infection of T cells through its HIV gp120-binding ability. The generation of a DC-SIGN-specific mAb has allowed us to determine that the acquisition of DC-SIGN expression during the monocyte-DC differentiation pathway is primarily induced by IL-4, and that GM-CSF cooperates with IL-4 to generate a high level of DC-SIGN mRNA and cell surface expression on immature MDDC. IL-4 was capable of inducing DC-SIGN expression on monocytes without affecting the expression of other MDDC differentiation markers. By contrast, IFN alpha, IFN-gamma, and TGF-beta were identified as negative regulators of DC-SIGN expression, as they prevented the IL-4-dependent induction of DC-SIGN mRNA on monocytes, and a similar inhibitory effect was exerted by dexamethasone, an inhibitor of the monocyte-MDDC differentiation pathway. The relevance of the inhibitory action of dexamethasone, IFN, and TGF-beta on DC-SIGN expression was emphasized by their ability to inhibit the DC-SIGN-dependent HIV-1 binding to differentiating MDDC. These results demonstrate that DC-SIGN, considered as a MDDC differentiation marker, is a molecule specifically expressed on IL-4-treated monocytes, and whose expression is subjected to a tight regulation by numerous cytokines and growth factors. This feature might help in the development of strategies to modulate the DC-SIGN-dependent cell surface attachment of HIV for therapeutic purposes. PMID- 11884428 TI - Thalidomide suppresses NF-kappa B activation induced by TNF and H2O2, but not that activated by ceramide, lipopolysaccharides, or phorbol ester. AB - Thalidomide ([+]-alpha-phthalimidoglutarimide), a psychoactive drug that readily crosses the blood-brain barrier, has been shown to exhibit anti-inflammatory, antiangiogenic, and immunosuppressive properties through a mechanism that is not fully established. Due to the central role of NF-kappaB in these responses, we postulated that thalidomide mediates its effects through suppression of NF-kappaB activation. We investigated the effects of thalidomide on NF-kappaB activation induced by various inflammatory agents in Jurkat cells. The treatment of these cells with thalidomide suppressed TNF-induced NF-kappaB activation, with optimum effect occurring at 50 microg/ml thalidomide. These effects were not restricted to T cells, as other hematopoietic and epithelial cell types were also inhibited. Thalidomide suppressed H(2)O(2)-induced NF-kappaB activation but had no effect on NF-kappaB activation induced by PMA, LPS, okadaic acid, or ceramide, suggesting selectivity in suppression of NF-kappaB. The suppression of TNF-induced NF-kappaB activation by thalidomide correlated with partial inhibition of TNF-induced degradation of an inhibitory subunit of NF-kappaB (IkappaBalpha), abrogation of IkappaBalpha kinase activation, and inhibition of NF-kappaB-dependent reporter gene expression. Thalidomide abolished the NF-kappaB-dependent reporter gene expression activated by overexpression of TNFR1, TNFR-associated factor-2, and NF kappaB-inducing kinase, but not that activated by the p65 subunit of NF-kappaB. Overall, our results clearly demonstrate that thalidomide suppresses NF-kappaB activation specifically induced by TNF and H(2)O(2) and that this may contribute to its role in suppression of proliferation, inflammation, angiogenesis, and the immune system. PMID- 11884429 TI - Polycyclic aromatic hydrocarbons affect functional differentiation and maturation of human monocyte-derived dendritic cells. AB - Polycyclic aromatic hydrocarbons (PAHs) such as benzo(a)pyrene (BP) are environmental carcinogens exhibiting potent immunosuppressive properties. To determine the cellular bases of this immunotoxicity, we have studied the effects of PAHs on differentiation, maturation, and function of monocyte-derived dendritic cells (DC). Exposure to BP during monocyte differentiation into DC upon the action of GM-CSF and IL-4 markedly inhibited the up-regulation of markers found in DC such as CD1a, CD80, and CD40, without altering cell viability. Besides BP, PAHs such as dimethylbenz(a)anthracene and benzanthracene also strongly altered CD1a levels. Moreover, DC generated in the presence of BP displayed decreased endocytic activity. Features of LPS-mediated maturation of DC, such as CD83 up-regulation and IL-12 secretion, were also impaired in response to BP treatment. BP-exposed DC poorly stimulated T cell proliferation in mixed leukocyte reactions compared with their untreated counterparts. In contrast to BP, the halogenated arylhydrocarbon 2,3,7,8-tetrachlorodibenzo-p-dioxin, which shares some features with PAHs, including interaction with the arylhydrocarbon receptor, failed to phenotypically alter differentiation of monocytes into DC, suggesting that binding to the arylhydrocarbon receptor cannot mimic PAH effects on DC. Overall, these data demonstrate that exposure to PAHs inhibits in vitro functional differentiation and maturation of blood monocyte-derived DC. Such an effect may contribute to the immunotoxicity of these environmental contaminants due to the major role that DC play as potent APC in the development of the immune response. PMID- 11884430 TI - L-selectin is not required for T cell-mediated autoimmune diabetes. AB - Administration of anti-L-selectin (CD62L) mAb to neonatal nonobese diabetic (NOD) mice mediates long term protection against the development of insulitis and overt diabetes. These results suggested that CD62L has a key role in the general function of beta cell-specific T cells. To further examine the role of CD62L in the development of type 1 diabetes, NOD mice lacking CD62L were established. The onset and frequency of overt diabetes were equivalent among CD62L(+/+), CD62L(+/ ), and CD62L(-/-) NOD littermates. Furthermore, patterns of T cell activation, migration, and beta cell-specific reactivity were similar in NOD mice of all three genotypes. Adoptive transfer experiments with CD62L(-/-) CD4(+) T cells prepared from BDC2.5 TCR transgenic mice revealed no apparent defects in migration to pancreatic lymph nodes, proliferation in response to beta cell Ag, or induction of diabetes in NOD.scid recipients. In conclusion, CD62L expression is not essential for the development of type 1 diabetes in NOD mice. PMID- 11884431 TI - Efficient and qualitatively distinct MHC class I-restricted presentation of antigen targeted to the endoplasmic reticulum. AB - For most nascent glycoprotein Ags, the MHC class I-restricted processing pathway begins in the endoplasmic reticulum (ER). From this location, they are translocated to the cytosol for degradation by the proteasome. A reasonable assumption is that processing of exocytic Ags is less efficient than that of cytosolic Ags, due to the requirement for additional handling, but that the processing pathways for the two types of proteins are otherwise similar. To test this, we compared the presentation of three epitopes within influenza nucleoprotein (NP) when this Ag is targeted to the cytosol or the ER. Surprisingly, under conditions of limited Ag expression, presentation of two proteasome-dependent epitopes is comparable when NP is targeted to the ER while presentation of a third is negatively impacted. Furthermore, presentation of the third epitope is unaffected by the addition of proteasome inhibitor when cytosolic NP is expressed but is significantly enhanced when exocytic NP is expressed. These results indicate that delivery of Ag to the ER need not preclude efficient presentation and that processing of cytosolic and ER-targeted Ag is qualitatively distinct. PMID- 11884432 TI - Cross-linking surface Ig delays CD40 ligand- and IL-4-induced B cell Ig class switching and reveals evidence for independent regulation of B cell proliferation and differentiation. AB - T cells stimulate B cells to divide and differentiate by providing activating signals in the form of inducible membrane-bound molecules and secreted cytokines. Provision of these signals in vitro reproduces many of the consequences of T-B collaboration in the absence of any form of Ag stimulation. Although clearly not obligatory, Ag signals appear to play an important regulatory role in numerous aspects of the B cell response. To examine directly the effect of an Ag signal, naive B cells were stimulated in the presence of rCD40 ligand, with or without IL 4 in the presence or absence of different anti-Ig mAbs. Anti-Ig mAbs exerted variable effects on the B cell division rate, from enhancement to no effect to inhibition. In contrast, all anti-Ig mAbs tested inhibited division-linked isotype switching to IgG1 and IgE. Thus, B cell Ag receptor ligands could modify the rates of B cell expansion and class switching independently. The ability of anti-Ig reagents to modify class switching suggests the B cell Ag receptor may play an important role in the selection of Ig isotypes during T cell-dependent humoral immune responses to Ags of different physical structure. PMID- 11884433 TI - Recognition of nonclassical HLA class I antigens by gamma delta T cells during pregnancy. AB - The healthy trophoblast does not express classical HLA-A and HLA-B products; therefore, an MHC-restricted recognition of trophoblast-presented Ags is unlikely. In the decidua and also in peripheral blood of healthy pregnant women, gammadelta T cells significantly increase in number. We investigated the possible role of gammadelta T cells in recognition of trophoblast-presented Ags. PBL and isolated gammadelta T cells from healthy pregnant women as well as from those at risk for premature pregnancy termination were conjugated to choriocarcinoma cells (JAR) transfected with nonclassical HLA Ags (HLA-E, HLA-G). To investigate the involvement of killer-inhibitory/killer-activatory receptors in trophoblast recognition, we tested the effect of CD94 block on cytotoxic activity of Vdelta2(+) enriched gammadelta T cells to HLA-E- and/or HLA-G-transfected targets. Lymphocytes from healthy pregnant women preferentially recognized HLA(-) choriocarcinoma cells, whereas those from pathologically pregnant patients did not discriminate between HLA(+) and HLA(-) cells. Normal pregnancy Vdelta2(+) T cells conjugated at a significantly increased rate to HLA-E transfectants, whereas Vdelta2(+) lymphocytes from pathologically pregnant women did not show a difference between those and HLA(-) cells. Blocking of the CD94 molecule of Vdelta2(+) lymphocytes from healthy pregnant women resulted in an increased cytotoxic activity to HLA-E-transfected target cells. These data indicate that Vdelta2(+) lymphocytes of healthy pregnant women recognize HLA-E on the trophoblast, whereas Vdelta1 cells react with other than HLA Ags. In contrast to Vdelta2(+) lymphocytes from healthy pregnant women, those from women with pathological pregnancies do not recognize HLA-E via their killer-inhibitory receptors and this might account for their high cytotoxic activity. PMID- 11884434 TI - IL-4 and T cells are required for the generation of IgG1 isotype antibodies against cardiolipin. AB - Infection with Mycobacterium tuberculosis induces Abs against a vast array of mycobacterial lipids and glycolipids. One of the most prominent lipid Ags recognized is cardiolipin (CL). The kinetics of the generation of anti-CL Abs during infection reveals that IgM titers to CL increase over time. Interestingly, at day 30 postinfection CL-specific IgG1 appears, an isotype usually dependent on T cell help. Using an immunization schedule with CL/anti-CL Ab complexes, which induces antiphospholipid syndrome in mice, we show that the generation of IgG1 to CL requires IL-4 and that optimal production is T cell dependent. IgG1 production to CL was impaired in nude (nu/nu) mice devoid in conventional T cells, but was not affected in mice deficient for either alphabeta TCR(+), gammadelta TCR(+), CD4(+), CD8(+), or NK1.1(+) T cells. We conclude that IgG1 production to CL depends on T cell help and IL-4, which can be provided by different T cell populations. This is the first report that IL-4 is indispensable for the induction of IgG1 Abs to lipid Ags. PMID- 11884435 TI - Impaired precursor B cell differentiation in Bruton's tyrosine kinase-deficient mice. AB - Bruton's tyrosine kinase (Btk) is a cytoplasmic signaling molecule that is crucial for precursor (pre-B) cell differentiation in humans. In this study, we show that during the transition of large cycling to small resting pre-B cells in the mouse, Btk-deficient cells failed to efficiently modulate the expression of CD43, surrogate L chain, CD2, and CD25. In an analysis of the kinetics of pre-B cell differentiation in vivo, Btk-deficient cells manifested a specific developmental delay within the small pre-B cell compartment of about 3 h, when compared with wild-type cells. Likewise, in in vitro bone marrow cultures, Btk deficient large cycling pre-B cells showed increased IL-7 mediated expansion and reduced developmental progression into noncycling CD2(+)CD25(+) surrogate L chain negative small pre-B cells and subsequently into Ig-positive B cells. Furthermore, the absence of Btk resulted in increased proliferative responses to IL-7 in recombination-activating gene-1-deficient pro-B cells. These findings identify a novel role for Btk in the regulation of the differentiation stage specific modulation of IL-7 responsiveness in pro-B and pre-B cells. Moreover, our results show that Btk is critical for an efficient transit through the small pre-B cell compartment, thereby regulating cell surface phenotype changes during the developmental progression of cytoplasmic mu H chain expressing pre-B cells into immature IgM(+) B cells. PMID- 11884436 TI - Differential in vivo persistence of two subsets of memory phenotype CD8 T cells defined by CD44 and CD122 expression levels. AB - The existence of distinct subsets of memory CD8 T cells with different characteristics is now well established. In this work, we describe two subsets of mouse CD8 T cells with memory characteristics that coexist in primed thymectomized TCR-transgenic F5 mice and that share some properties with the human central and effector memory cells. The first subset corresponds to CD8 T cells generated following nucleoprotein 68 peptide priming which are CD44(int)CD122(-)nucleoprotein 68/H-2D(b) tetramer(+) and express high levels of CCR7 mRNA. In contrast, CD8 T cells in the second subset are CD44(high)CD122(+), are heterogeneous in terms of Ag specificity, and express low levels of CCR7 mRNA. We have studied the functional characteristics and the persistence of these two subsets in thymectomized mice. CD44(int) CD8 T cells persist like naive cells; i.e., they are slowly lost with time. However, surviving cells maintain their phenotype and memory characteristics for the entire life span of the animal. In contrast, CD44(high) CD8 T cells are persistent and accumulate in thymectomized but not euthymic mice. This is correlated with an increased in vivo proliferative and survival potential of these cells. These results show that acquisition of enhanced functional characteristics and long-term persistence by memory T cells are independent. This may have important consequences for the design of specific vaccine. PMID- 11884437 TI - BCR engagement induces Fas resistance in primary B cells in the absence of functional Bruton's tyrosine kinase. AB - B cell susceptibility to Fas-mediated apoptosis is regulated in a receptor specific fashion. CD40 engagement produces marked sensitivity to Fas killing, whereas surface Ig (sIg) engagement blocks Fas signaling for cell death in otherwise sensitive, CD40-stimulated B cell targets, and thus, induces a state of Fas resistance. The signaling mediator, Bruton's tyrosine kinase (Btk), is required for certain sIg-triggered responses, and Btk is reported to directly bind Fas and block Fas-mediated apoptosis. For these reasons, the role of Btk as a mediator of sIg-induced Fas resistance was examined. Dysfunction of Btk through mutation, and absence of Btk through deletion did not interfere with induction of Fas resistance by anti-Ig. This may be due, at least in part, to induction of Btk dependent Bcl-2 family members by anti-Ig after CD40 ligand treatment. However, the susceptibility to Fas-mediated apoptosis of B cell targets stimulated by CD40 ligand alone was increased in the absence of Btk. These results indicate that Fas resistance produced by sIg triggering does not require Btk, but suggests that in certain situations Btk modulates B cell susceptibility to Fas killing. PMID- 11884438 TI - T cell immunity to lymphoma following treatment with anti-CD40 monoclonal antibody. AB - In this study we demonstrate that treatment with anti-CD40 mAb eradicates a range of mouse lymphomas (BCL(1), A31, A20, and EL4), but only when used against i.v. tumor doses in excess of 10(7) cells. Only partial protection was seen against smaller tumor loads. We saw no evidence that anti-CD40 mAb changed the phenotype of the lymphomas or inhibited their growth in the initial period following treatment, but it did result in a rapid expansion of cytotoxic CD8(+) cells that was able to clear the neoplastic disease and provide long-term protection against tumor rechallenge. The CTL responses were blocked by mAb against a range of coreceptors and cytokines, including CD8, B7-1, B7-2, LFA-1, and IFN-gamma, but not CD4 or CTLA-4, indicating the presence of a conventional cellular Th1 response. Furthermore, we found evidence of cross-recognition between lymphomas (BCL(1) and A20) as measured by cytotoxicity and IFN-gamma responses in vitro and using tumor rechallenge experiments, suggesting common target Ags. Finally, although anti-CD40 was shown to stimulate NK cell killing, we could find no role for these cells in controlling tumor growth. These data underline the ability of anti-CD40 mAb to potentiate CTL responses and the potency of cellular immunity in eradicating large quantities of syngeneic tumor. PMID- 11884439 TI - CD28 costimulation mediates down-regulation of p27kip1 and cell cycle progression by activation of the PI3K/PKB signaling pathway in primary human T cells. AB - CD28 provides a costimulatory signal that cooperates with the TCR/CD3 complex to induce T cell activation, cytokine production, and clonal expansion. We have recently shown that CD28 directly regulates progression of T lymphocytes through the cell cycle. Although a number of signaling pathways have been linked to the TCR/CD3 and to CD28, it is not known how these two receptors cooperate to induce cell cycle progression. Here, using cell-permeable pharmacologic inhibitors of phosphatidylinositol 3-hydroxykinase (PI3K) and mitogen-activated protein kinase kinase (MEK1/2), we show that cell cycle progression of primary T lymphocytes requires simultaneous activation of PI3K- and MEK1/2-dependent pathways. Decreased abundance of cyclin-dependent kinase inhibitor p27(kip1), which requires simultaneous TCR/CD3 and CD28 ligation, was dependent upon both MEK and PI3K activity. Ligation of TCR/CD3, but not CD28 alone, resulted in activation of MEK targets extracellular signal-related kinase 1/2, whereas ligation of CD28 alone was sufficient for activation of PI3K target protein kinase B (PKB; c-Akt). CD28 ligation alone was also sufficient to mediate inactivating phosphorylation of PKB target glycogen synthase kinase-3 (GSK-3). Moreover, direct inactivation of GSK-3 by LiCl in the presence of anti-CD3, but not in the presence of anti CD28, resulted in down-regulation of p27(kip1), hyperphosphorylation of retinoblastoma tumor suppressor gene product, and cellular proliferation. Thus, inactivation of the PI3K-PKB target GSK-3 could substitute for CD28 but not for CD3 signals. These results show that the PI3K-PKB pathway links CD28 to cell cycle progression and suggest that p27(kip1) integrates mitogenic MEK- and PI3K dependent signals from TCR and CD28 in primary T lymphocytes. PMID- 11884440 TI - Lipid microdomain clustering induces a redistribution of antigen recognition and adhesion molecules on human T lymphocytes. AB - The study of lipid microdomains in the plasma membrane is a topic of recent interest in leukocyte biology. Many T cell activation and signaling molecules are found to be associated with lipid microdomains and have been implicated in normal T cell function. It has been proposed that lipid microdomains with their associated molecules move by lateral diffusion to areas of cellular interactions to initiate signaling pathways. Using sucrose density gradients we have found that human T cell beta(1) integrins are not normally associated with lipid microdomains. However, cross-linking of GM1 through cholera toxin B-subunit (CTB) causes an enrichment of beta(1) integrins in microdomain fractions, suggesting that cross-linking lipid microdomains causes a reorganization of molecular associations. Fluorescent microscopy was used to examine the localization of various lymphocyte surface molecules before and after lipid microdomain cross linking. Lymphocytes treated with FITC-CTB reveal an endocytic vesicle that is enriched in TCR and CD59, while beta(1) integrin, CD43, and LFA-3 were not localized in the vesicle. However, when anti-CTB Abs are used to cross-link lipid microdomains, the microdomains are not internalized but are clustered on the cell surface. In this study, CD59, CD43, and beta(1) integrin are all seen to colocalize in a new lipid microdomain from which LFA-3 remains excluded and the TCR is now dissociated. These findings show that cross-linking lipid microdomains can cause a dynamic rearrangement of the normal order of T lymphocyte microdomains into an organization where novel associations are created and signaling pathways may be initiated. PMID- 11884441 TI - Focal localization of placental protein 14 toward sites of TCR engagement. AB - TCR signal transduction is amplified by the dynamic accumulation of accessory molecules at APC-T cell contact sites, along with the simultaneous exclusion from these sites of negative regulators, such as certain tyrosine phosphatases and large glycosylated proteins. However, given the general nature of the cytoskeleton-driven clustering mechanism underlying molecular segregation events at the APC-T cell interaction site, the possibility exists that negative regulators might similarly be segregated at these sites. Using fluorescence microscopy, we have demonstrated that placental protein 14 (PP14), a direct T cell inhibitor, focuses toward APC-T cell contact sites in conjunction with conjugate formation. We have further established that the function of PP14 is dependent upon its localization to the sites of TCR triggering, where it negatively regulates T cell activation. Thus, PP14 provides an example of a soluble negative T cell regulator whose inhibitory activity is linked to modulation of the APC-T cell contact site, thereby hindering early events triggered by the TCR. PMID- 11884442 TI - Characterization of the Fas ligand/Fas-dependent apoptosis of antiretroviral, class I MHC tetramer-defined, CD8+ CTL by in vivo retrovirus-infected cells. AB - C57BL/6 (B6; H-2(b)) mice mount strong AKR/Gross murine leukemia virus (MuLV) specific CD8(+) CTL responses to the immunodominant K(b)-restricted epitope, KSPWFTTL, of endogenous AKR/Gross MuLV. In sharp contrast, spontaneous virus expressing AKR.H-2(b) congenic mice are low/nonresponders for the generation of AKR/Gross MuLV-specific CTL. Furthermore, when viable AKR.H-2(b) spleen cells are cocultured with primed responder B6 antiviral precursor CTL, the AKR.H-2(b) cells function as "veto" cells that actively mediate the inhibition of antiviral CTL generation. AKR.H-2(b) veto cell inhibition is virus specific, MHC restricted, contact dependent, and mediated through veto cell Fas ligand/responder T cell Fas interactions. In this study, following specific priming and secondary in vitro restimulation, antiretroviral CD8(+) CTL were identified by a labeled K(b)/KSPWFTTL tetramer and flow cytometry, enabling direct visualization of AKR.H 2(b) veto cell-mediated depletion of these CTL. A 65-93% reduction in the number of B6 K(b)/KSPWFTTL tetramer(+) CTL correlated with a similar reduction in antiviral CTL cytotoxicity. Addition on sequential days to the antiviral CTL restimulation cultures of either 1) AKR.H-2(b) veto cells or 2) a blocking Fas-Ig fusion protein (to cultures also containing AKR.H-2(b) veto cells) to block inhibition demonstrated that AKR.H-2(b) veto cells begin to inhibit B6 precursor CTL/CTL expansion during days 2 and 3 of the 6-day culture. Shortly thereafter, a high percentage of B6 tetramer(+) CTL cocultured with AKR.H-2(b) veto cells was annexin V positive and Fas(high), indicating apoptosis as the mechanism of veto cell inhibition. Experiments using the irreversible inhibitor emetine demonstrated that AKR.H-2(b) cells had to be metabolically active and capable of protein synthesis to function as veto cells. Of the tetramer-positive CTL that survived veto cell-mediated apoptosis, there was no marked skewing from the preferential usage of Vbeta4, 8.1/8.2, and 11 TCR normally observed. These findings provide further insight into the complexity of host/virus interactions and suggest a fail-safe escape mechanism by virus-infected cells for epitopes residing in critical areas of viral proteins that cannot accommodate variations of amino acid sequence. PMID- 11884443 TI - CD40 ligand functions non-cell autonomously to promote deletion of self-reactive thymocytes. AB - CD40 ligand (CD40L)-deficient mice have been shown to have a defect in negative selection of self-reactive T cells during thymic development. However, the mechanism by which CD40L promotes deletion of autoreactive thymocytes has not yet been elucidated. We have studied negative selection in response to endogenous superantigens in CD40L-deficient mice and, consistent with previous reports, have found a defect in negative selection in these mice. To test the requirement for expression of CD40L on T cells undergoing negative selection, we have generated chimeric mice in which CD40L wild-type and CD40L-deficient thymocytes coexist. We find that both CD40L wild-type and CD40L-deficient thymocytes undergo equivalent and efficient negative selection when these populations coexist in chimeric mice. These results indicate that CD40L can function in a non-cell-autonomous manner during negative selection. Deletion of superantigen-reactive thymocytes was normal in B7-1/B7-2 double-knockout mice, indicating that CD40-CD40L-dependent negative selection is not solely mediated by B7 up-regulation and facilitation of B7-dependent T cell signaling. Finally, although the absence of CD40-CD40L interactions impairs negative selection of autoreactive CD4(+) and CD8(+) cells during thymic development, we find that self-reactive T cells are deleted in the mature CD4(+) population through a CD40L-independent pathway. PMID- 11884444 TI - Impact of antigen presentation on TCR modulation and cytokine release: implications for detection and sorting of antigen-specific CD8+ T cells using HLA A2 wild-type or HLA-A2 mutant tetrameric complexes. AB - Soluble MHC class I molecules loaded with antigenic peptides are available either to detect and to enumerate or, alternatively, to sort and expand MHC class I restricted and peptide-reactive T cells. A defined number of MHC class I/peptide complexes can now be implemented to measure T cell responses induced upon Ag specific stimulation, including CD3/CD8/zeta-chain down-regulation, pattern, and quantity of cytokine secretion. As a paradigm, we analyzed the reactivity of a Melan-A/MART-1-specific and HLA-A2-restricted CD8(+) T cell clone to either soluble or solid-phase presented peptides, including the naturally processed and presented Melan-A/MART-1 peptide AAGIGILTV or the peptide analog ELAGIGILTV presented either by the HLA-A2 wild-type (wt) or mutant (alanineright arrowvaline aa 245) MHC class I molecule, which reduces engagement of the CD8 molecule with the HLA-A2 heavy chain. Soluble MHC class I complexes were used as either monomeric or tetrameric complexes. Soluble monomeric MHC class I complexes, loaded with the Melan-A/MART-1 peptide, resulted in CD3/CD8 and TCR zeta-chain down-regulation, but did not induce measurable cytokine release. In general, differences pertaining to CD3/CD8/zeta-chain regulation and cytokine release, including IL-2, IFN-gamma, and GM-CSF, were associated with 1) the format of Ag presentation (monomeric vs tetrameric MHC class I complexes), 2) wt vs mutant HLA A2 molecules, and 3) the target Ag (wt vs analog peptide). These differences are to be considered if T cells are exposed to recombinant MHC class I Ags loaded with peptides implemented for detection, activation, or sorting of Ag-specific T cells. PMID- 11884445 TI - Essential role for CD40 ligand interactions in T lymphocyte-mediated modulation of the murine immune response to pneumococcal capsular polysaccharides. AB - Protection against infection with pneumococci is provided by anti-capsular polysaccharide (caps-PS) Abs. We investigated whether CD40 ligand (CD40L) plays a role in T lymphocyte-mediated regulation of the immune response to caps-PS, which are considered thymus-independent Ags. Administration of MR1, an antagonist mAb against murine CD40L, in BALB/c mice immunized with Pneumovax resulted in an inhibition of the IgM and IgG Ab response for various caps-PS serotypes. Evidence for the involvement of CD4(+) T lymphocytes in the Ab response to caps-PS was obtained in SCID/SCID mice that, when reconstituted with B lymphocytes and CD4(+) T lymphocytes, mounted a higher specific IgM response compared with SCID/SCID mice reconstituted with only B lymphocytes. This helper effect of CD4(+) T lymphocytes was abrogated by MR1. Blocking CD40L in vitro decreased the IgM response to caps-PS and abolished the helper effect of CD4(+) T lymphocytes. CD8(+) T lymphocyte-depleted murine spleen cells mounted a higher in vivo immune response than total murine spleen cells, which provided evidence for a suppressive role of CD8(+) T lymphocytes on the anti-caps-PS immune response. CD4(+) T lymphocyte-depleted murine spleen cells, leaving a B and CD8(+) T lymphocyte fraction, elicited only a weak in vivo and in vitro Ab response, which was enhanced after MR1 administration. In summary, our data provide evidence that T lymphocytes contribute to the regulation of the anti-caps-PS immune response in a CD40L-dependent manner. PMID- 11884446 TI - Complement receptor type 1 (CD35) mediates inhibitory signals in human B lymphocytes. AB - The complement system---particularly component C3---has been demonstrated to be a key link between innate and adaptive immunity. The trimolecular complex of complement receptor type 2 (CR2), CD19, and CD81 is known to promote B cell activation when coligated with the B cell Ag receptor. In the present study, we aimed to elucidate the role of human complement receptor type 1 (CR1), the other C3-receptor on B cells. As ligand, aggregated C3 and aggregated C3(H(2)O), i.e., multimeric "C3b-like C3", are used, which bind to CR1, but not to CR2. In experiments studying the functional consequences of CR1-clustering, the multimeric ligand is shown to inhibit the proliferation of tonsil B cells activated with a suboptimal dose of anti-IgM F(ab')(2). Importantly, this inhibitory activity also occurs in the presence of the costimulatory cytokines IL 2 and IL-15. The anti-IgM-induced transient increase in the concentration of intracellular free Ca(2+) and phosphorylation of several cytoplasmic proteins are strongly reduced in the presence of the CR1 ligand. Data presented indicate that CR1 has a negative regulatory role in the B cell Ag receptor mediated activation of human B lymphocytes. PMID- 11884447 TI - A novel murine model of Graves' hyperthyroidism with intramuscular injection of adenovirus expressing the thyrotropin receptor. AB - In this work we report a novel method to efficiently induce a murine model of Graves' hyperthyroidism. Inbred mice of different strains were immunized by i.m. injection with adenovirus expressing thyrotropin receptor (TSHR) or beta galactosidase (1 x 10(11) particles/mouse, three times at 3-wk intervals) and followed up to 8 wk after the third immunization. Fifty-five percent of female and 33% of male BALB/c (H-2(d)) and 25% of female C57BL/6 (H-2(b)) mice developed Graves'-like hyperthyroidism with elevated serum thyroxine (T(4)) levels and positive anti-TSHR autoantibodies with thyroid-stimulating Ig (TSI) and TSH binding inhibiting Ig (TBII) activities. In contrast, none of female CBA/J (H 2(k)), DBA/1J (H-2(q)), or SJL/J (H-2(s)) mice developed Graves' hyperthyroidism or anti-TSHR autoantibodies except SJL/J, which showed strong TBII activities. There was a significant positive correlation between TSI values and T(4) levels, but the correlations between T(4) and TBII and between TSI and TBII were very weak. TSI activities in sera from hyperthyroid mice measured with some chimeric TSH/lutropin receptors suggested that their epitope(s) on TSHR appeared similar to those in patients with Graves' disease. The thyroid glands from hyperthyroid mice displayed diffuse enlargement with hypertrophy and hypercellularity of follicular epithelia with occasional protrusion into the follicular lumen, characteristics of Graves' hyperthyroidism. Decreased amounts of colloid were also observed. However, there was no inflammatory cell infiltration. Furthermore, extraocular muscles from hyperthyroid mice were normal. Thus, the highly efficient means that we now report to induce Graves' hyperthyroidism in mice will be very useful for studying the pathogenesis of autoimmunity in Graves' disease. PMID- 11884448 TI - Regulation of cytokine expression by ligands of peroxisome proliferator activated receptors. AB - Peroxisome proliferator activated receptors (PPARs) are ligand-activated transcription factors with diverse actions including adipocyte differentiation and lipid metabolism. Recent studies have revealed anti-inflammatory activities, but the majority of these studies have been performed in monocyte/macrophages. In these studies, we investigate the effects of PPAR ligands in murine mitogen activated splenocytes. Ciglitazone, a PPARgamma ligand, consistently decreased IFN-gamma and IL-2 production by mitogen-activated splenocytes and had modest effects on splenocyte proliferation. The effects of WY14,643, a representative of the fibrate class of PPARalpha ligands, on splenocyte proliferation and IL-2 levels are less marked than those observed with the PPARgamma ligand. In addition, treatment with WY14,643 and other fibrates led to marked increases in supernatant concentrations of IL-4. However, treatment with a potent and specific PPARalpha ligand (GW7,647) did not augment IL-4. Also, WY14,643 induced IL-4 expression in splenocytes from PPARalpha knockout mice, suggesting that the fibrate effect on IL-4 was largely through a PPARalpha-independent mechanism. This increase in IL-4 was associated with and causatively related to augmented expression of CD23 by CD45R/B220(+) cells. We also demonstrate that PPARgamma gene expression is up-regulated in T cells by mitogen activation, that it is positively regulated by IL-4 and WY14,643, and that it is blocked by anti-IL-4. Finally, we demonstrate that WY14,643 can modestly augment IL-4 promoter activity in a PPARalpha-independent manner. In concert, these findings support the roles of PPAR ligands in modulating inflammatory responses involving lymphocytes but also establish potent effects of the fibrate class of PPARalpha ligands on IL-4 expression that are receptor independent. PMID- 11884449 TI - CD66a interactions between human melanoma and NK cells: a novel class I MHC independent inhibitory mechanism of cytotoxicity. AB - NK cells are able to kill virus-infected and tumor cells via a panel of lysis receptors. Cells expressing class I MHC proteins are protected from lysis primarily due to the interactions of several families of NK receptors with both classical and nonclassical class I MHC proteins. In this study we show that a class I MHC-deficient melanoma cell line (1106mel) is stained with several Ig fused lysis receptors, suggesting the expression of the appropriate lysis ligands. Surprisingly, however, this melanoma line was not killed by CD16 negative NK clones. The lack of killing is shown to be the result of homotypic CD66a interactions between the melanoma line and the NK cells. Furthermore, 721.221 cells expressing the CD66a protein were protected from lysis by YTS cells and by NK cells expressing the CD66a protein. Redirected lysis experiments demonstrated that the strength of the inhibitory effect is correlated with the levels of CD66a expression. Finally, the expression of CD66a protein was observed on NK cells derived from patients with malignant melanoma. These findings suggest the existence of a novel class I MHC-independent inhibitory mechanism of human NK cell cytotoxicity. This may be a mechanism that is used by some of the class I MHC-negative melanoma cells to evade attack by CD66a-positive NK cells. PMID- 11884450 TI - A role for CCR9 in T lymphocyte development and migration. AB - CCR9 mediates chemotaxis in response to CCL25/thymus-expressed chemokine and is selectively expressed on T cells in the thymus and small intestine. To investigate the role of CCR9 in T cell development, the CCR9 gene was disrupted by homologous recombination. B cell development, thymic alphabeta-T cell development, and thymocyte selection appeared unimpaired in adult CCR9-deficient (CCR9(-/-)) mice. However, competitive transplantation experiments revealed that bone marrow from CCR9(-/-) mice was less efficient at repopulating the thymus of lethally irradiated Rag-1(-/-) mice than bone marrow from littermate CCR9(+/+) mice. CCR9(-/-) mice had increased numbers of peripheral gammadelta-T cells but reduced numbers of gammadeltaTCR(+) and CD8alphabeta(+)alphabetaTCR(+) intraepithelial lymphocytes in the small intestine. Thus, CCR9 plays an important, although not indispensable, role in regulating the development and/or migration of both alphabeta(-) and gammadelta(-) T lymphocytes. PMID- 11884451 TI - Differential patterns of methylation of the IFN-gamma promoter at CpG and non-CpG sites underlie differences in IFN-gamma gene expression between human neonatal and adult CD45RO- T cells. AB - IFN-gamma is a potent pleiotropic Th1 cytokine, the production of which is tightly regulated during fetal development. Negative control of fetal/neonatal IFN-gamma production is generally attributed to the Th1-antagonistic effect of mediators produced by the placenta, but evidence exists of additional and more direct transcriptional regulation. We report that neonatal (cord blood) CD3(+)/CD45RO(-) T cells, in particular the CD4(+)/CD45RO(-) subset, are hypermethylated at CpG and non-CpG (CpA and CpT) sites within and adjacent to the IFN-gamma promoter. In contrast, CpG methylation patterns in cord blood IFN-gamma producing CD8(+)/CD45RO(-) T cells and CD56(+)/CD16(+)/CD3(-) NK cells did not differ significantly from those in their adult counterparts. Consistent with this finding, IFN-gamma production by stimulated naive cord blood CD4(+) T cells is reduced 5- to 10-fold relative to adult CD4(+) T cells, whereas production levels in neonatal and adult CD8(+) T cells are of a similar order. Evidence of significant CpA and CpT methylation was not discovered in promoter sequence from other cytokines (IL-4, TNF-alpha, or IFN-gammaR alpha-chain). We additionally demonstrate that overexpression of DNA methyltransferase 3a in embryonic kidney carcinoma cells is accompanied by CpA methylation of the IFN-gamma promoter. PMID- 11884452 TI - Dualism of oxidized lipoproteins in provoking and attenuating the oxidative burst in macrophages: role of peroxisome proliferator-activated receptor-gamma. AB - Activation and deactivation of macrophages are of considerable importance during the development of various disease states, atherosclerosis among others. Macrophage activation is achieved by oxidized lipoproteins (oxLDL) and is determined by oxygen radical (ROS) formation. The oxidative burst was measured by flow cytometry and quantitated by oxidation of the redox-sensitive dye dichlorodihydrofluorescein diacetate. Short-time stimulation dose-dependently elicited ROS formation. Diphenylene iodonium prevented ROS formation, thus pointing to the involvement of a NAD(P)H oxidase in producing reduced oxygen species. In contrast, preincubation of macrophages with oxLDL for 16 h showed an attenuated oxidative burst upon a second contact with oxLDL. Taking into account that oxLDL is an established peroxisome proliferator-activated receptor-gamma (PPARgamma) agonist and considering the anti-inflammatory properties of PPARgamma, we went on and showed that a PPARgamma agonist such as ciglitazone attenuated ROS formation. Along that line, major lipid peroxidation products of oxLDL, such as 9- and 13-hydroxyoctadecadienoic acid, shared that performance. Supporting evidence that PPARgamma activation accounted for reduced ROS generation came from studies in which proliferator-activated receptor response element decoy oligonucleotides, but not a mutated oligonucleotide, supplied in front of oxLDL delivery regained a complete oxidative burst upon cell activation. We conclude that oxLDL not only elicits an oxidative burst upon first contact, but also promotes desensitization of macrophages via activation of PPARgamma. Desensitization of macrophages may have important consequences for the behavior of macrophages/foam cells in atherosclerotic lesions. PMID- 11884453 TI - Two new isotype-specific switching activities detected for Ig class switching. AB - Ig class switch recombination (CSR) occurs by an intrachromosomal deletional process between switch (S) regions in B cells. To facilitate the study of CSR, we derived a new B cell line, 1.B4.B6, which is uniquely capable of mu --> gamma3, mu --> epsilon, and mu --> alpha, but not mu --> gamma1 CSR at its endogenous loci. The 1.B4.B6 cell line was used in combination with plasmid-based isotype specific S substrates in transient transfection assays to test for the presence of trans-acting switching activities. The 1.B4.B6 cell line supports mu --> gamma3, but not mu --> gamma1 recombination, on S substrates. In contrast, normal splenic B cells activated with LPS and IL-4 are capable of plasmid-based mu --> gamma1 CSR and demonstrate that this S plasmid is active. Activation-induced deaminase (AID) was used as a marker to identify existing B cell lines as possible candidates for supporting CSR. The M12 and A20 cell lines were identified as AID positive and, following activation with CD40L and other activators, were found to differentially support mu --> epsilon and mu --> alpha plasmid-based CSR. These studies provide evidence for two new switching activities for mu --> gamma1 and mu --> epsilon CSR, which are distinct from mu - > gamma3 and mu --> alpha switching activities previously described. AID is expressed in all the B cell lines capable of CSR, but cannot account for the isotype specificity defined by the S plasmid assay. These results are consistent with a model in which isotype-specific switching factors are either isotype specific recombinases or DNA binding proteins with sequence specificity for S DNA. PMID- 11884454 TI - The ubiquitously expressed DNA-binding protein late SV40 factor binds Ig switch regions and represses class switching to IgA. AB - Ig heavy chain class switch recombination (CSR) determines the expression of Ig isotypes. The molecular mechanism of CSR and the factors regulating this process have remained elusive. Recombination occurs primarily within switch (S) regions, located upstream of each heavy chain gene (except Cdelta). These repetitive sequences contain consensus DNA-binding sites for the DNA-binding protein late SV40 factor (LSF) (CP2/leader-binding protein-1c). In this study, we demonstrate by EMSA that purified rLSF, as well as LSF within B cell extracts, directly binds both Smu and Salpha sequences. To determine whether LSF is involved in regulating CSR, two different LSF dominant negative variants were stably expressed in the mouse B cell line I.29 mu, which can be induced to switch from IgM to IgA. Overexpression of these dominant negative LSF proteins results in decreased levels of endogenous LSF DNA-binding activity and an increase in cells undergoing CSR. Thus, LSF represses class switching to IgA. In agreement, LSF DNA-binding activity was found to decrease in whole cell extracts from splenic B cells induced to undergo class switching. To elucidate the mechanism of CSR regulation by LSF, the interactions of LSF with proteins involved in chromatin modification were tested in vitro. LSF interacts with both histone deacetylases and the corepressor Sin3A. We propose that LSF represses CSR by histone deacetylation of chromatin within S regions, thereby limiting accessibility to the switch recombination machinery. PMID- 11884455 TI - The Pmed1 gene promoter of human Fc gamma RIIIA can function as a NK/T cell specific restriction element, which involves binding of Sp1 transcription factor. AB - The low-affinity receptor for IgG (human FcgammaRIIIA) is selectively expressed by a subset of T lymphocytes, NK cells, and macrophages. To understand the mechanisms underlying this pattern of cell type-specific expression, we initially identified alternative promoters, Pmed1/2 and Pprox, in the 5' end of the FcgammaRIIIA gene. In this study, we focused on the Pmed1 promoter and demonstrated this 93-bp region to be highly specific in governing restriction to NK/T cell lines. This property of Pmed1 is context independent and can extend to a disparate promoter. Deletion analysis defined a contribution of two separate elements located to the 5' 21-bp (-942/-922) and 3' 72-bp (-921/-850) regions of Pmed1 in conferring NK/T cell specificity. The 5' part of Pmed1 contains binding sites for Sp1 and NK element-recognizing factors and substitution mapping studies revealed a critical requirement of the Sp1-I site. The importance of Sp1 protein to regulate maximal Pmed1 promoter activity was further established by EMSAs and cotransfection experiments in Sp1-null Drosophila SL2 cells. Our data suggest that Sp1 can contribute, in part, to NK/T cell restriction and further indicate that the FcgammaRIIIA Pmed1 sequence might be useful to direct the NK/T cell specific expression of heterologous genes. PMID- 11884456 TI - Phosphorylation and O-linked glycosylation of Elf-1 leads to its translocation to the nucleus and binding to the promoter of the TCR zeta-chain. AB - Elf-1, a member of the E 26-specific transcription factor family with a predicted molecular mass of 68 kDa, is involved in the transcriptional regulation of several hematopoietic cell genes. We demonstrate that Elf-1 exists primarily as a 98-kDa form in the nucleus and as an 80-kDa form in the cytoplasm. Phosphorylation and O-linked glycosylation contribute to the increased posttranslational molecular mass of Elf-1. The 98-kDa Elf-1 is released from the cytoplasm tethering retinoblastoma protein and moves to the nucleus, where it binds to the promoter of the TCR zeta-chain gene. Finally, the cytoplasmic 98-kDa form enters the proteasome pathway and undergoes degradation. In conclusion, different forms of Elf-1 are the products of posttranslational modifications that determine its subcellular localization, activity, and metabolic degradation. PMID- 11884458 TI - Critical role for activation of antigen-presenting cells in priming of cytotoxic T cell responses after vaccination with virus-like particles. AB - Virus-like particles (VLPs) are known to induce strong Ab responses in the absence of adjuvants. In addition, VLPs are able to prime CTL responses in vivo. To study the efficiency of this latter process, we fused peptide p33 derived from lymphocytic choriomeningitis virus to the hepatitis B core Ag, which spontaneously assembles into VLPs (p33-VLPs). These p33-VLPs were efficiently processed in vitro and in vivo for MHC class I presentation. Nevertheless, p33 VLPs induced weak CTL responses that failed to mediate effective protection from viral challenge. However, if APCs were activated concomitantly in vivo using either anti-CD40 Abs or CpG oligonucleotides, the CTL responses induced were fully protective against infection with lymphocytic choriomeningitis virus or recombinant vaccinia virus. Moreover, these CTL responses were comparable to responses generally induced by live vaccines, because they could be measured in primary ex vivo (51)Cr release assays. Thus, while VLPs alone are inefficient at inducing CTL responses, they become very powerful vaccines if applied together with substances that activate APCs. PMID- 11884457 TI - Optimal T cell responses to Cryptococcus neoformans mannoprotein are dependent on recognition of conjugated carbohydrates by mannose receptors. AB - Cryptococcosis is a leading cause of death among individuals with compromised T cell function. Soluble Cryptococcus neoformans mannoproteins (MP) have emerged as promising vaccine candidates due to their capacity to elicit delayed-type hypersensitivity and Th type 1-like cytokines, both critical to the clearance of this pathogenic yeast. In this study, the mechanisms responsible for the potent immunostimulatory properties of MP were explored. Using Chinese hamster ovary cells expressing human macrophage mannose receptor (MMR), we determined that MP is a MMR ligand. Functionally, competitive blockade of multilectin mannose receptors (MR) on APCs diminished MP-dependent stimulation of primary T cells from immunized mice and the MP-reactive CD4(+) T cell hybridoma, P1D6, by 72 and 99%, respectively. Removal of O-linked saccharides from MP by beta-elimination inhibited MP-dependent stimulation of P1D6 and primary T cells by 89 and 90%, respectively. In addition, MP-dependent stimulation of P1D6 was abrogated after digestion with proteinase K, suggesting the protein core of MP contributed the antigenic moiety presented by APC. Stimulation of P1D6 by MP also was abolished using APC obtained from invariant chain-deficient mice, demonstrating Ag presentation was MHC class II restricted. Our data suggest that MP is a ligand for the MMR and that T cell stimulation is functionally inhibited either by competitive blockade of MR or by removal of carbohydrate residues critical for recognition. The demonstration that efficient T cell responses to MP require recognition of terminal mannose groups by MMR provides both a molecular basis for the immunogenicity of cryptococcal MP and support for vaccination strategies that target MR. PMID- 11884459 TI - Eotaxin/CCL11 suppresses IL-8/CXCL8 secretion from human dermal microvascular endothelial cells. AB - The CC chemokine eotaxin/CCL11 is known to bind to the receptor CCR3 on eosinophils and Th2-type lymphocytes. In this study, we demonstrate that CCR3 is expressed on a subpopulation of primary human dermal microvascular endothelial cells and is up-regulated by TNF-alpha. We found that incubation of human dermal microvascular endothelial cells with recombinant eotaxin/CCL11 suppresses TNF alpha-induced production of the neutrophil-specific chemokine IL-8/CXCL8. The eotaxin/CCL11-suppressive effect on endothelial cells was not seen on IL-1beta induced IL-8/CXCL8 release. Eotaxin/CCL11 showed no effect on TNF-alpha-induced up-regulation of growth-related oncogene-alpha or IFN-gamma-inducible protein-10, two other CXC chemokines tested, and did not affect production of the CC chemokines monocyte chemoattractant protein-1/CCL2 and RANTES/CCL5, or the adhesion molecules ICAM-1 and E-selectin. These results suggest that eotaxin/CXCL11 is not effecting a general suppression of TNF-alphaR levels or signal transduction. Suppression of IL-8/CXCL8 was abrogated in the presence of anti-CCR3 mAb, pertussis toxin, and wortmannin, indicating it was mediated by the CCR3 receptor, G(i) proteins, and phosphatidylinositol 3-kinase signaling. Eotaxin/CCL11 decreased steady state levels of IL-8/CXCL8 mRNA in TNF-alpha stimulated cells, an effect mediated in part by an acceleration of IL-8 mRNA decay. Eotaxin/CCL11 may down-regulate production of the neutrophil chemoattractant IL-8/CXCL8 by endothelial cells in vivo, acting as a negative regulator of neutrophil recruitment. This may play an important biological role in the prevention of overzealous inflammatory responses, aiding in the resolution of acute inflammation or transition from neutrophilic to mononuclear/eosinophilic inflammation. PMID- 11884460 TI - HIV-1 down-modulates gamma signaling chain of Fc gamma R in human macrophages: a possible mechanism for inhibition of phagocytosis. AB - HIV-1 infection impairs a number of macrophage effector functions, thereby contributing to development of opportunistic infections and the pathogenesis of AIDS. FcgammaR-mediated phagocytosis by human monocyte-derived macrophages (MDM) is inhibited by HIV-1 infection in vitro, and the underlying mechanism was investigated in this study. Inhibition of phagocytosis directly correlated with the multiplicity of HIV-1 infection. Expression of surface FcgammaRs was unaffected by HIV-1 infection, suggesting that inhibition of phagocytosis occurred during or after receptor binding. HIV-1 infection of MDM markedly inhibited tyrosine phosphorylation of the cellular proteins, which occurs following engagement of FcgammaRs, suggesting a defect downstream of initial receptor activation. FcgammaR-mediated phagocytosis in HIV-infected MDM was associated with inhibition of phosphorylation of tyrosine kinases from two different families, Hck and Syk, defective formation of Syk complexes with other tyrosine-phosphorylated proteins, and inhibition of paxillin activation. Down modulation of protein expression but not mRNA of the gamma signaling subunit of FcgammaR (a docking site for Syk) was observed in HIV-infected MDM. Infection of MDM with a construct of HIV-1 in which nef was replaced with the gene for the gamma signaling subunit augmented FcgammaR-mediated phagocytosis, suggesting that down-modulation of gamma-chain protein expression in HIV-infected MDM caused the defective FcgammaR-mediated signaling and impairment of phagocytosis. This study is the first to demonstrate a specific alteration in phagocytosis signal transduction pathway, which provides a mechanism for the observed impaired FcgammaR-mediated phagocytosis in HIV-infected macrophages and contributes to the understanding of how HIV-1 impairs cell-mediated immunity leading to HIV-1 disease progression. PMID- 11884461 TI - Dendritic cells pulsed with fungal RNA induce protective immunity to Candida albicans in hematopoietic transplantation. AB - Immature myeloid dendritic cells (DC) phagocytose yeasts and hyphae of the fungus Candida albicans and induce different Th cell responses to the fungus. Ingestion of yeasts activates DC for production of IL-12 and Th1 priming, while ingestion of hyphae induces IL-4 production and Th2 priming. In vivo, generation of antifungal protective immunity is induced upon injection of DC ex vivo pulsed with Candida yeasts but not hyphae. In the present study we sought to determine the functional activity of DC transfected with yeast or hyphal RNA. It was found that DC, from either spleens or bone marrow, transfected with yeast, but not hyphal, RNA 1) express fungal mannoproteins on their surface; 2) undergo functional maturation, as revealed by the up-regulated expression of MHC class II Ags and costimulatory molecules; 3) produce IL-12 but no IL-4; 4) are capable of inducing Th1-dependent antifungal resistance when delivered s.c. in vivo in nontransplanted mice; and 5) provide protection against the fungus in allogeneic bone marrow-transplanted mice, by accelerating the functional recovery of Candida specific IFN-gamma-producing CD4(+) donor lymphocytes. These results indicate the efficacy of DC pulsed with Candida yeasts or yeast RNA as fungal vaccines and point to the potential use of RNA-transfected DC as anti-infective vaccines in conditions that negate the use of attenuated microorganisms or in the case of poor availability of protective Ags. PMID- 11884462 TI - Macrophage effector functions controlled by Bruton's tyrosine kinase are more crucial than the cytokine balance of T cell responses for microfilarial clearance. AB - Macrophages from X-linked immunodeficient (xid) mice lacking functional Bruton's tyrosine kinase (Btk) show poor NO induction and enhanced IL-12 induction, and contribute to delayed clearance of injected microfilaria (mf) in vivo. We now show that Btk is involved in other macrophage effector functions, such as bactericidal activity and secretion of proinflammatory cytokines (TNF-alpha, IL 1beta), but not the T cell-directed cytokine IL-12. Induction of some transcriptional regulators of the NF-kappaB family, crucial for the expression of proinflammatory cytokines, is also poor in Btk-deficient macrophages. Thus, Btk appears to be involved in signaling for inducible effector functions, but not APC functions, in macrophages. Furthermore, adoptive transfer of T cells from mf infected xid or wild-type mice did not alter the course of mf clearance in recipients, mf clearance was unaltered in IFN-gamma-deficient mice, and improved mf clearance was seen only if greater inducibility of IL-12 was accompanied by greater NO secretion from macrophages, as seen in Ity(r) C.D2 mice as compared with congenic Ity(s) BALB/c mice. Thus, delayed mf clearance in xid mice was correlated not with the high IL-12/Th1 phenotype but with low NO induction levels. Also, xid macrophages showed poor toxicity to mf in vitro as compared with wild-type macrophages. Inhibition of NO production blocked this mf cytotoxicity, and an NF-kappaB inhibitor blocked both NO induction and mf cytotoxicity. Thus, Btk is involved in inducing many macrophage effector functions, and delayed mf clearance seen in Btk-deficient xid mice is due to poor NO induction in macrophages, resulting in compromised microfilarial toxicity. PMID- 11884463 TI - Recruitment kinetics and composition of antibody-secreting cells within the central nervous system following viral encephalomyelitis. AB - Infection by the neurotropic JHM strain of mouse hepatitis virus produces an acute demyelinating encephalomyelitis. While cellular immunity initially eliminates infectious virus, CNS viral persistence is predominantly controlled by humoral immunity. To better understand the distinct phases of immune control within the CNS, the kinetics of humoral immune responses were determined in infected mice. Early during clearance of the JHM strain of mouse hepatitis virus, only few virus-specific Ab-secreting cells (ASC) were detected in the periphery or CNS, although mature B cells and ASC without viral specificity were recruited into the CNS concomitant with T cells. Serum antiviral Ab and CNS virus-specific ASC became prominent only during final elimination of infectious virus. Virus specific ASC peaked in lymphoid organs before the CNS, suggesting peripheral B cell priming and maturation. Following elimination of infectious virus, virus specific ASC continued to increase within the CNS and then remained stable during persistence, in contrast to declining T cell numbers. These data comprise three novel findings. Rapid recruitment of B cells in the absence of specific Ab secretion supports a potential Ab-independent effector function involving lysis of virus-infected cells. Delayed recruitment relative to viral clearance and subsequent maintenance of a stable CNS ASC population demonstrate differential regulation of T and B lymphocytes within the infected CNS. This supports a critical role of humoral immunity in regulating viral CNS persistence. Lastly, altered antiviral ASC specificities following clearance of infectious virus suggest ongoing recruitment of peripheral memory cells and/or local B cell differentiation. PMID- 11884464 TI - Protection against influenza virus infection in polymeric Ig receptor knockout mice immunized intranasally with adjuvant-combined vaccines. AB - The role of secretory IgA in conferring cross-protective immunity was examined in polymeric (p)IgR knockout (KO) mice immunized intranasally with different inactivated vaccines prepared from A/PR/8/34 (H1N1), A/Yamagata/120/86 (H1N1), A/Beijing/262/95 (H1N1), and B/Ibaraki/2/85 viruses and infected with the A/PR/8/34 virus in the upper respiratory tract (RT)-restricting volume. In wild type mice, immunization with A/PR/8/34 or its variant (A/Yamagata/120/86 and A/Beijing/262/95) vaccines conferred complete protection or partial cross protection against infection, while the B-type virus vaccine failed to provide protection. The protection or cross-protection was accompanied by an increase in the nasal A/PR/8/34 hemagglutinin-reactive IgA concentration, which was estimated to be >30 times the serum IgA concentration and much higher than the nasal IgG concentration. In contrast, the blockade of transepithelial transport of dimeric IgA in pIgR-KO mice reduced the degree of protection or cross-protection, in parallel with the marked increase in serum IgA concentration and the decrease in nasal IgA concentration (about 20 and 0.3 times those in wild-type mice, respectively). The degree of the reduction of protection or cross-protection was moderately reversed by the low but non-negligible level of nasal IgA, transudates from the accumulated serum IgA. These results, together with the absence of the IgA-dependent cross-protection in the lower RT and the unaltered level of nasal or serum IgG in wild-type and pIgR-KO mice, confirm that the actively secreted IgA plays an important role in cross-protection against variant virus infection in the upper RT, which cannot be substituted by serum IgG. PMID- 11884465 TI - Toll-like receptor 4-MD-2 complex mediates the signal transduction induced by flavolipin, an amino acid-containing lipid unique to Flavobacterium meningosepticum. AB - Flavolipin, an amino acid-containing lipid isolated from Flavobacterium meningosepticum, induces many immune responses. It has been shown that flavolipin does not induce an immune response of macrophages derived from C3H/HeJ mice, which possess a point mutation in Toll-like receptor 4 (TLR4). To determine whether TLR4 or the molecular complex of TLR4 and TLR4 association molecule MD-2 mediates the flavolipin signal, flavolipin responsiveness was examined by measuring NF-kappaB activation in Ba/F3 cells and Ba/F3 transfectants expressing TLR4 or both TLR4 and MD-2. Flavolipin-induced NF-kappaB activation was detected in the cells expressing both TLR4 and MD-2, but not in the other cells. Expression of CD14 in the transfectant expressing both TLR4 and MD-2 increased the sensitivity to flavolipin. Furthermore, flavolipin stereoisomers were chemically synthesized, and their abilities to induce NF-kappaB activation were examined. (R)-Flavolipin, in which the configuration of the lipid moiety is R, induced NF-kappaB activation via the TLR4-MD-2 complex, but (S)-flavolipin did not. In this study, we demonstrated the involvement of TLR4-MD-2 and CD14 in flavolipin signaling and the importance of the (R)-configuration of the flavolipin lipid moiety for the induction of an immune response via TLR4-MD-2. PMID- 11884466 TI - The role of IFN in respiratory syncytial virus pathogenesis. AB - Formalin-inactivated respiratory syncytial virus (RSV) vaccine preparations have been shown to cause enhanced disease in naive hosts following natural infection. In this study we demonstrate a similar pattern of enhanced disease severity following primary RSV infection of IFN-nonresponsive STAT1(-/-) mice. STAT1(-/-) mice showed markedly increased illness compared with wild-type BALB/c animals following RSV inoculation despite similar lung virus titers and rates of virus clearance. Histologically, STAT1(-/-) animals had eosinophilic and neutrophilic pulmonary infiltrates not present in wild-type or IFN-gamma(-/-)-infected mice. In cytokine analyses of infected lung tissue, IFN-gamma was induced in both STAT1(-/-) and wild-type mice, with preferential IL-4, IL-5, and IL-13 induction only in the STAT1(-/-) animals. Eotaxin was detected in the lungs of both wild type and STAT1(-/-) mice following infection, with a 1.7-fold increase over wild type in the STAT1(-/-) mice. Using a peptide epitope newly identified in the RSV fusion protein, we were able to demonstrate that wild-type memory CD4(+) T cells stimulated by this peptide produce primarily IFN-gamma, while STAT1(-/-)CD4(+) cells produce primarily IL-13. These findings suggest that STAT1 activation by both type I (alphabeta) and type II (gamma) IFNs plays an important role in establishing a protective, Th1 Ag-specific immune response to RSV infection. PMID- 11884467 TI - IL-13-induced chemokine responses in the lung: role of CCR2 in the pathogenesis of IL-13-induced inflammation and remodeling. AB - IL-13 stimulates inflammatory and remodeling responses and contributes to the pathogenesis of human airways disorders. To further understand the cellular and molecular events that mediate these responses, we characterized the effects of IL 13 on monocyte chemotactic proteins (MCPs) and compared the tissue effects of transgenic IL-13 in mice with wild-type (+/+) and null (-/-) CCR2 loci. Transgenic IL-13 was a potent stimulator of MCP-1, -2, -3, and -5. This stimulation was not specific for MCPs because macrophage-inflammatory protein (MIP)-1alpha, MIP-1beta, MIP-2, MIP-3alpha, thymus- and activation-regulated chemokine, thymus-expressed chemokine, eotaxin, eotaxin 2, macrophage-derived chemokines, and C10 were also induced. The ability of IL-13 to increase lung size, alveolar size, and lung compliance, to stimulate pulmonary inflammation, hyaluronic acid accumulation, and tissue fibrosis, and to cause respiratory failure and death were markedly decreased, whereas mucus metaplasia was not altered in CCR2(-/-) mice. CCR2 deficiency did not decrease the basal or IL-13 stimulated expression of target matrix metalloproteinases or cathepsins but did increase the levels of mRNA encoding alpha1-antitrypsin, tissue inhibitor of metalloproteinase-1, -2, and -4, and secretory leukocyte proteinase inhibitor. In addition, the levels of bioactive and total TGF-beta(1) were decreased in lavage fluids from IL-13 transgenic mice with -/- CCR2 loci. These studies demonstrate that IL-13 is a potent stimulator of MCPs and other CC chemokines and document the importance of MCP-CCR2 signaling in the pathogenesis of the IL-13-induced pulmonary phenotype. PMID- 11884468 TI - Role of lipopolysaccharide-binding protein in early alcohol-induced liver injury in mice. AB - Cellular responses to endotoxins are enhanced markedly by LPS-binding protein (LBP). Furthermore, it has been demonstrated that endotoxins and proinflammatory cytokines such as TNF-alpha participate in early alcohol-induced liver injury. Therefore, in this study, a long-term intragastric ethanol feeding model was used to test the hypothesis that LBP is involved in alcoholic hepatitis by comparing LBP knockout and wild-type mice. Two-month-old female mice were fed a high-fat liquid diet with either ethanol or isocaloric maltose-dextrin as control continuously for 4 wk. There was no difference in mean urine alcohol concentrations between the groups fed ethanol. Dietary alcohol significantly increased liver to body weight ratios and serum alanine aminotransferase levels in wild-type mice (189 +/- 31 U/L) over high-fat controls (24 +/- 7 U/L), effects which were blunted significantly in LBP knockout mice (60 +/- 17 U/L). Although no significant pathological changes were observed in high-fat controls, 4 wk of dietary ethanol caused steatosis, mild inflammation, and focal necrosis in wild type animals as expected (pathology score, 5.9 +/- 0.5). These pathological changes were reduced significantly in LBP knockout mice fed ethanol (score, 2.6 +/- 0.5). Endotoxin levels in the portal vein were increased significantly after 4 wk in both groups fed ethanol. Moreover, ethanol increased TNF-alpha mRNA expression in wild-type, but not in LBP knockout mice. These data are consistent with the hypothesis that LBP plays an important role in early alcohol-induced liver injury by enhancing LPS-induced signal transduction, most likely in Kupffer cells. PMID- 11884469 TI - The cutaneous reverse Arthus reaction requires intercellular adhesion molecule 1 and L-selectin expression. AB - The deposition of immune complexes (IC) induces an acute inflammatory response with tissue injury. IC-induced inflammation is mediated by inflammatory cell infiltration, a process highly regulated by expression of multiple adhesion molecules. To assess the role of L-selectin and ICAM-1 in this pathogenetic process, the cutaneous reverse passive Arthus reaction was examined in mice lacking L-selectin (L-selectin(-/-)), ICAM-1 (ICAM-1(-/-)), or both (L selectin/ICAM-1(-/-)). Edema and hemorrhage, which peaked 4 and 8 h after IC challenge, respectively, were significantly reduced in L-selectin(-/-), ICAM-1(-/ ), and L-selectin/ICAM-1(-/-) mice compared with wild-type littermates. In general, edema and hemorrhage were more significantly inhibited in ICAM-1(-/-) mice than in L-selectin(-/-) mice, but were most significantly reduced in L selectin/ICAM-1(-/-) mice compared with ICAM-1(-/-) or L-selectin(-/-) mice. Decreased edema and hemorrhage correlated with reduced neutrophil and mast cell infiltration in all adhesion molecule-deficient mice, but leukocyte infiltration was most affected in L-selectin/ICAM-1(-/-) mice. Reduced neutrophil and mast cell infiltration was also observed for all mutant mice in the peritoneal Arthus reaction. Furthermore, cutaneous TNF-alpha production was inhibited in each deficient mouse, which paralleled the reductions in cutaneous inflammation. These results indicate that ICAM-1 and L-selectin cooperatively contribute to the cutaneous Arthus reaction by regulating neutrophil and mast cell recruitment and suggest that ICAM-1 and L-selectin are therapeutic targets for human IC-mediated disease. PMID- 11884470 TI - Activation of nuclear orphan receptor NURR1 transcription by NF-kappa B and cyclic adenosine 5'-monophosphate response element-binding protein in rheumatoid arthritis synovial tissue. AB - Modulation of the NURR subfamily of nuclear receptors may be an important mechanism regulating pathways associated with inflammatory joint disease. We examined the signaling mechanisms through which inflammatory mediators, produced by rheumatoid arthritis (RA) synovial tissue, contribute to the regulation of the NURR subfamily. Markedly enhanced expression of NURR1 is observed in synovial tissue of patients with RA compared with normal subjects. Modulation by proinflammatory mediators in primary RA and normal synoviocytes shows that PGE(2), IL-1beta, and TNF-alpha markedly enhance NURR1 mRNA and protein levels in contrast to other subfamily members, NUR77 and NOR-1. We have established that transcriptional activation of the NURR1 gene by IL-1beta and TNF-alpha requires a proximal promoter region that contains a consensus NF-kappaB DNA-binding motif. IL-1beta- and TNF-alpha-induced NF-kappaB binding to this site is due predominantly to p65-p50 heterodimer and p50 homodimer subunit protein complexes. We further demonstrate a direct CREB-1-dependent regulation by PGE(2) situated at promoter region -171/-163. Moreover, analyses confirm the presence of CREB-1 and NF-kappaB p50 and p65 subunit binding to the NURR1 promoter under basal conditions in freshly explanted RA synovial tissue. In summary, enhanced NF kappaB- and CREB-1-binding activity on the NURR1 promoter by inflammatory mediators delineates novel mechanisms in the regulation of NURR1 transcription. PGE(2)-, TNF-alpha-, and IL-1beta-dependent stimulation of the NURR1 gene implies that NURR1 induction represents a point of convergence of at least two distinct signaling pathways, suggesting an important common role for this transcription factor in mediating multiple inflammatory signals. PMID- 11884471 TI - Lamina propria CD4+ T lymphocytes synergize with murine intestinal epithelial cells to enhance proinflammatory response against an intracellular pathogen. AB - Acute and lethal ileitis can be elicited in certain strains of inbred mice after oral infection with the intracellular protozoan parasite Toxoplasma gondii. The development of this inflammatory process is dependent upon the induction of a robust Th1 response, including overproduction of IFN-gamma, TNF-alpha, and NO, as has been reported in other experimental models of human inflammatory bowel disease. In this study we have investigated the role of CD4(+) T cells from the lamina propria (LP) in the early inflammatory events after T. gondii infection using isolated and primary cultured intestinal cells from infected mice and immortalized mouse mIC(cl2) intestinal epithelial cells. Primed LP CD4(+) T cells isolated from parasite-infected mice produce substantial quantities of both IFN gamma and TNF-alpha. IFN-gamma- and TNF-alpha-producing LP CD4(+) T cells synergize with infected mIC(cl2) and enhance the production of several inflammatory chemokines including macrophage-inflammatory protein-2, monocyte chemoattractant protein-1, monocyte chemoattractant protein-3, macrophage inflammatory protein-1alphabeta, and IFN-gamma-inducible protein-10. Furthermore, primed LP CD4(+) T cells cocultured with infected mIC(cl2) inhibited replication of the parasite in the intestinal epithelial cells. Thus, LP CD4(+) T cells can interact with parasite-infected intestinal epithelial cells and alter the expression of several proinflammatory products that have been associated with the development of intestinal inflammation. The interaction between these two components of the gut mucosal compartment (CD4(+) T cells and enterocytes) may play a role in the immunopathogenesis of this pathogen-driven experimental inflammatory bowel disease model. PMID- 11884472 TI - Heat shock proteins gp96 and hsp70 activate the release of nitric oxide by APCs. AB - NO is a cytotoxic and immunomodulatory cytokine produced by macrophages and dendritic cells. We show that stimulation of murine and human macrophages with the heat shock proteins gp96 and hsp70 results in induction of inducible NO synthase and the production of NO. The release of NO by monocytes exposed to hsp60 has been documented previously. Immature, but not mature, dendritic cells respond in the same manner. The activity of heat shock proteins is relatively unaffected by an antagonist of LPS, and is abrogated by heat denaturation. Macrophages have been shown previously to produce NO in response to stimulation with IFN-gamma; stimulation of macrophages with mixtures of IFN-gamma and gp96 or hsp70 leads to a synergistic production of NO. The present observations extend the roles of these heat shock proteins in innate immune responses to another potent and highly conserved function of APC. PMID- 11884473 TI - Critical role for T cells in Sephadex-induced airway inflammation: pharmacological and immunological characterization and molecular biomarker identification. AB - Intratracheal instillation of Sephadex particles is a convenient model for assessing the impact of potential anti-inflammatory compounds on lung eosinophilia thought to be a key feature in asthma pathophysiology. However, the underlying cellular and molecular mechanisms involved are poorly understood. We have studied the time course of Sephadex-induced lung eosinophilia, changes in pulmonary T cell numbers, and gene and protein expression as well as the immunological and pharmacological modulation of these inflammatory indices in the Sprague Dawley rat. Sephadex increased T cell numbers (including CD4(+) T cells) and evoked a pulmonary eosinophilia that was associated with an increase in gene/protein expression of the Th2-type cytokines IL-4, IL-5, and IL-13 and eotaxin in lung tissue. Sephadex instillation also induced airway hyperreactivity to acetylcholine and bradykinin. A neutralizing Ab (R73) against the alphabeta TCR caused 54% depletion of total (CD2(+)) pulmonary T cells accompanied by a significant inhibition of IL-4, IL-13 and eotaxin gene expression together with suppression (65% inhibition) of eosinophils in lung tissue 24 h after Sephadex treatment. Sephadex-induced eosinophilia and Th2 cytokine gene and/or protein expression were sensitive to cyclosporin A and budesonide, compounds that inhibit T cell function, suggesting a pivotal role for T cells in orchestrating Sephadex induced inflammation in this model. PMID- 11884474 TI - Pathogenesis of murine experimental allergic rhinitis: a study of local and systemic consequences of IL-5 deficiency. AB - Recent studies have demonstrated an important role for IL-5-dependent bone marrow eosinophil progenitors in allergic inflammation. However, studies using anti-IL-5 mAbs in human asthmatics have failed to suppress lower airway hyperresponsiveness despite suppression of eosinophilia; therefore, it is critical to examine the role of IL-5 and bone marrow responses in the pathogenesis of allergic airway disease. To do this, we studied the effects of IL-5 deficiency (IL-5(-/-)) on bone marrow function as well as clinical and local events, using an established experimental murine model of allergic rhinitis. Age-matched IL-5(+/+) and IL-5(-/ ) BALB/c mice were sensitized to OVA followed by 2 wk of daily OVA intranasal challenge. IL-5(-/-) OVA-sensitized mice had significantly higher nasal mucosal CD4(+) cells and basophilic cell counts as well as nasal symptoms and histamine hyperresponsiveness than the nonsensitized group; however, there was no eosinophilia in either nasal mucosa or bone marrow; significantly lower numbers of eosinophil/basophil CFU and maturing CFU eosinophils in the presence of recombinant mouse IL-5 in vitro; and significantly lower expression of IL-5Ralpha on bone marrow CD34(+)CD45(+) progenitor cells in IL-5(-/-) mice. These findings suggest that IL-5 is required for normal bone marrow eosinophilopoiesis, in response to specific Ag sensitization, during the development of experimental allergic rhinitis. However, the results also suggest that suppression of the IL-5 eosinophil pathway in this model of allergic rhinitis may not completely suppress clinical symptoms or nasal histamine hyperresponsiveness, because of the existence of other cytokine-progenitor pathways that may induce and maintain the presence of other inflammatory cell populations. PMID- 11884475 TI - Antimicrobial peptides initiate IL-1 beta posttranslational processing: a novel role beyond innate immunity. AB - Human monocytes stimulated with LPS produce large quantities of prointerleukin 1beta, but little of this cytokine product is released extracellularly as the mature biologically active species. To demonstrate efficient proteolytic cleavage and export, cytokine-producing cells require a secondary effector stimulus. In an attempt to identify agents that may serve as initiators of IL-1beta posttranslational processing in vivo, LPS-activated human monocytes were treated with several individual antimicrobial peptides. Two peptides derived from porcine neutrophils, protegrin (PTG)-1 and PTG-3, promoted rapid and efficient release of mature IL-1beta. The PTG-mediated response engaged a mechanism similar to that initiated by extracellular ATP acting via the P2X(7) receptor. Thus, both processes were disrupted by a caspase inhibitor, both were sensitive to ethacrynic acid and CP-424,174, two pharmacological agents that suppress posttranslational processing, and both were negated by elevation of extracellular potassium. Moreover, the PTGs, like ATP, promoted a dramatic change in monocyte morphology and a loss of membrane latency. The PTG response was concentration dependent and was influenced profoundly by components within the culture medium. In contrast, porcine neutrophil antimicrobial peptides PR-26 and PR-39 did not initiate IL-1beta posttranslational processing. The human defensin HNP-1 and the frog peptide magainin 1 elicited export of 17-kDa IL-1beta, but these agents were less efficient than PTGs. As a result of this ability to promote release of potent proinflammatory cytokines such as IL-1beta, select antimicrobial peptides may possess important immunomodulatory functions that extend beyond innate immunity. PMID- 11884476 TI - The Helicobacter pylori blood group antigen-binding adhesin facilitates bacterial colonization and augments a nonspecific immune response. AB - Presence of the Helicobacter pylori adherence factor blood group Ag-binding adhesin (BabA; binding to Lewis(b) (Le(b))) is associated with ulcer disease, adenocarcinoma, and precancerous lesions. The importance of BabA for bacterial colonization and the inflammatory response is unknown. A total of 141 antral biopsies from H. pylori-infected patients were assessed in regard to the degree of granulocytic (G0 degrees--G3 degrees) and lymphocytic (L1 degrees--L3 degrees) infiltration. DNA genotypes of babA2 (the transcriptionally active gene of BabA), cagA, and vacAs1/2 were determined by PCR. Colonization density and Le(b) status on gastric epithelial cells were determined by immunohistochemistry. Real-time quantitative (TaqMan) RT-PCR determined mRNA expression of IL-8, TNF -alpha, and the Th1 markers IFN-gamma and the IL-12R beta2 chain. A total of 91% of infected patients were Le(b) positive. The vacAs1(+)/cagA(+) strains harboring babA2 showed significantly higher levels of granulocytic infiltration, bacterial colonization, and IL-8 mRNA than vacAs1(+)/cagA(+) strains lacking babA2. IL-8 mRNA and protein production by KATO III cells in vitro increased dose dependently with addition of different numbers of type 1 strains (G27 and 2808 strains, 0.1- 20 bacteria/cell). The mRNA expression of TNF-alpha, IFN-gamma, and IL-12R beta2 was higher in H. pylori-positive patients than in controls, but it did not differ significantly between patients infected with different strain types. These data suggest that BabA facilitates colonization of H. pylori and thereby increases IL 8 response, resulting in enhanced mucosal inflammation. Infection with strains harboring BabA thereby augment a nonspecific immune response, whereas the Th1 response toward H. pylori appears to be independent of BabA, cytotoxin-associated gene A, or vacuolating cytotoxin. PMID- 11884477 TI - A novel susceptibility locus on chromosome 2 in the (New Zealand Black x New Zealand White)F1 hybrid mouse model of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is inherited as a complex polygenic trait. (New Zealand Black (NZB) x New Zealand White (NZW)) F(1) hybrid mice develop symptoms that remarkably resemble human SLE, but (NZB x PL/J)F(1) hybrids do not develop lupus. Our study was conducted using (NZW x PL/J)F(1) x NZB (BWP) mice to determine the effects of the PL/J and the NZW genome on disease. Forty-five percent of BWP female mice had significant proteinuria and 25% died before 12 mo of age compared with (NZB x NZW)F(1) mice in which >90% developed severe renal disease and died before 12 mo. The analysis of BWP mice revealed a novel locus (chi(2) = 25.0; p < 1 x 10(-6); log of likelihood = 6.6 for mortality) designated Wbw1 on chromosome 2, which apparently plays an important role in the development of the disease. We also observed that both H-2 class II (the u haplotype) and TNF alpha (TNF(z) allele) appear to contribute to the disease. A suggestive linkage to proteinuria and death was found for an NZW allele (designated Wbw2) telomeric to the H-2 locus. The NZW allele that overlaps with the previously described locus Sle1c at the telomeric part of chromosome 1 was associated with antinuclear autoantibody production in the present study. Furthermore, the previously identified Sle and Lbw susceptibility loci were associated with an increased incidence of disease. Thus, multiple NZW alleles including the Wbw1 allele discovered in this study contribute to disease induction, in conjunction with the NZB genome, and the PL/J genome appears to be protective. PMID- 11884478 TI - A 320-kilobase artificial chromosome encoding the human HLA DR3-DQ2 MHC haplotype confers HLA restriction in transgenic mice. AB - MHC class II haplotypes control the specificity of Th immune responses and susceptibility to many autoimmune diseases. Understanding the role of HLA class II haplotypes in immunity is hampered by the lack of animal models expressing these genes as authentic cis-haplotypes. In this study we describe transgenic expression of the autoimmune prone HLA DR3-DQ2 haplotype from a yeast artificial chromosome (YAC) containing an intact similar320-kb region from HLA DRA to DQB2. In YAC-transgenic mice HLA DR and DQ gene products were expressed on B cells, macrophages, and dendritic cells, but not on T cells indicating cell-specific regulation. Positive selection of the CD4 compartment by human class II molecules was 67% efficient in YAC-homozygous mice lacking endogenous class II molecules (Abeta(null/null)) and expressing only murine CD4. A broad range of TCR Vbeta families was used in the peripheral T cell repertoire, which was also purged of Vbeta5-, Vbeta11-, and Vbeta12-bearing T cells by endogenous mouse mammary tumor virus-encoded superantigens. Expression of the HLA DR3-DQ2 haplotype on the Abeta(null/null) background was associated with normal CD8-dependent clearance of virus from influenza-infected mice and development of CD4-dependent protection from otherwise lethal infection with Salmonella typhimurium. HLA DR- and DQ restricted T cell responses were also elicited following immunization with known T cell determinants presented by these molecules. These findings demonstrate the potential for human MHC class II haplotypes to function efficiently in transgenic mice and should provide valuable tools for developing humanized models of MHC associated autoimmune diseases. PMID- 11884479 TI - Interaction of antibodies to proteinase 3 (classic anti-neutrophil cytoplasmic antibody) with human renal tubular epithelial cells: impact on signaling events and inflammatory mediator generation. AB - Among the anti-neutrophil cytoplasmic Abs (ANCA), those targeting proteinase 3 (PR3) have a high sensitivity and specificity for Wegener's granulomatosis (WG). A pathogenetic role for these autoantibodies has been proposed due to their capacity of activating neutrophils in vitro. Recently, PR3 was also detected in human renal tubular epithelial cells (TEC). In the present study, the effect of murine monoclonal anti-PR3 Abs (anti-PR3) and purified c-ANCA targeting PR3 from WG serum on isolated human renal tubular cell signaling and inflammatory mediator release was characterized. Priming of TEC with TNF-alpha resulted in surface expression of PR3, as quantified in immunofluorescence studies and by flow cytometry. Moreover, PR3 was immunoprecipitated on surface-labeled TEC. Primed TEC responded to anti-PR3 with a dose- and time-dependent activation of phosphoinositide hydrolysis, resulting in a remarkable accumulation of inositolphosphates. Control IgG was entirely ineffective, whereas PR3-ANCA reproduced the phosphoinositide response. The signaling response was accompanied by a pronounced release of superoxidanion into the cell supernatant. Moreover, large amounts of PGE(2) and, to a lesser extent, of thromboxane B(2), the stable metabolite of TxA(2), were secreted from anti-PR3-stimulated TEC. In parallel, a rise in intracellular cAMP levels was observed, which was blocked by the cyclooxygenase inhibitor indomethacin. We conclude that anti-PR3 Abs directly target renal TECs, thereby provoking pronounced activation of the phosphoinositide-related signal transduction pathway. Associated metabolic events such as the release of reactive oxygen species and lipid mediators may directly contribute to the development of renal lesions and loss of kidney function in WG. PMID- 11884480 TI - TNF-TNFR2 interactions are critical for the development of intestinal graft versus-host disease in MHC class II-disparate (C57BL/6J-->C57BL/6J x bm12)F1 mice. AB - TNF-TNFR2 interactions promote MHC class II-stimulated alloresponses while TNF TNFR1 interactions promote MHC class I-stimulated alloresponses. The present studies were designed to evaluate whether TNF-TNFR2 interactions were involved in the in vivo generation of CD4(+) T cell-mediated intestinal graft-versus-host disease (GVHD) in the (C57BL/6J (hereafter called B6) --> B6 x B6.C-H-2(bm12) (bm12))F(1) GVHD model. Briefly, 5 x 10(6) splenic CD4(+) T lymphocytes from B6.TNFR2(-/-) or control B6 mice were transferred with 1--2 x 10(6) T cell depleted B6 bone marrow cells (BMC) to irradiated MHC class II-disparate (bm12 x B6)F(1) mice. Weight loss, intestinal inflammation, and the surface expression of CD45RB (memory marker) on intestinal and splenic lymphocytes were assessed. IL-2 and IFN-alpha mRNA levels in intestinal lymphocytes were assessed by nuclease protection assays. A significant reduction in weight loss and intestinal inflammation was observed in recipients of the TNFR2(-/-)CD4(+) SpC. Similarly, a significant decrease was noted in T cell numbers and in CD45RB(low) (activated/memory) expression on intestinal but not CD4(+) T cells in recipients of TNFR2(-/-)CD4(+) spleen cells. IL-2 and IFN-alpha mRNA levels were reduced in the intestine in the recipients of TNFR2(-/-) splenic CD4(+) T cells. These results indicate that TNF-TNFR2 interactions are important for the development of intestinal inflammation and activation/differentiation of Th1 cytokine responses by intestinal lymphocytes in MHC class II-disparate GVHD while playing an insignificant role in donor T cell activation in the spleen. PMID- 11884481 TI - Alpha-actinin is a cross-reactive renal target for pathogenic anti-DNA antibodies. AB - Anti-DNA Abs commonly found in patients with systemic lupus erythematosus are thought to play an important pathogenic role in lupus nephritis. Anti-DNA Abs may contribute to renal disease by cross-reactivity with renal Ags, the identity of which remain elusive. To identify a target Ag for pathogenic anti-DNA Abs, we performed Western blotting and immunoprecipitations of mesangial cell lysates from the lupus-prone MRL-lpr/lpr mouse and a nonautoimmune BALB/c mouse with the pathogenic anti-DNA Ab R4A. We found that R4A (but not a nonpathogenic Ab mutant of R4A) binds to and immunoprecipitates a 100-kDa protein expressed on the cell surface and in lysates of MRL-lpr/lpr mesangial cells. DNase treatment of the lysate and of the R4A Ab did not effect binding, indicating that the binding of R4A to the 100-kDa protein was direct and not mediated by an antigenic bridge containing DNA. Binding was greatly diminished in BALB/c lysates, suggesting that Ag expression or availability at the level of the target organ may be a factor in determining susceptibility to lupus nephritis. Following identification of this 100-kDa protein as nonmuscle alpha-actinin, binding of R4A to alpha-actinin was confirmed by Western blot, ELISA, inhibition studies, and immunofluorescence. High titers of anti-alpha-actinin Abs were present in sera and kidney eluates of lupus mice with active nephritis. These results indicate that the nephritogenicity of some anti-DNA Abs may be mediated via cross-reactivity with alpha-actinin. Furthermore, variations in target Ag display between individuals may underlie differential susceptibility to anti-DNA Ab-induced renal disease. PMID- 11884482 TI - Skin inflammation during contact hypersensitivity is mediated by early recruitment of CD8+ T cytotoxic 1 cells inducing keratinocyte apoptosis. AB - Contact hypersensitivity (CHS) is a T cell-mediated, Ag-specific skin inflammation induced by skin exposure to haptens in sensitized individuals. Th1/T cytotoxic 1 cells are effector cells of CHS, whereas Th2/T regulatory CD4(+) T cells have down-regulating properties. We have previously shown that CHS to 2,4 dinitrofluorobenzene is mediated by specific CD8(+) effector cells, whose cytolytic activity is mandatory for induction of skin inflammation. In this study, using immunohistochemistry and RT-PCR analysis, we show that CD8(+) T cells are rapidly recruited into the skin at the site of hapten challenge before the onset of clinical and histological signs of skin inflammation. This early CD8(+) T cell recruitment is concomitant with: 1) transient IFN-gamma mRNA expression suggesting local activation of effector cells; and 2) induction of keratinocyte (KC) apoptosis which gradually increased to a maximum at the peak of the CHS response. Alternatively, skin infiltration of CD4(+) T cells occurred later and coincided with the peak of the CHS reaction and the beginning of the resolution of skin inflammation. Mice deficient in CD8(+) T cells did not develop CHS, whereas mice deficient in CD4(+) T cells developed an enhanced inflammatory response with increased numbers of CD8(+) T cells recruited in the skin associated with massive KC apoptosis. These data show that CHS is due to the early and selective recruitment in the skin of CD8(+) T cytotoxic 1 effector cells responsible for KC apoptosis. PMID- 11884484 TI - A novel approach to the analysis of specificity, clonality, and frequency of HIV specific T cell responses reveals a potential mechanism for control of viral escape. AB - Escape from the CD8(+) T cell response through epitope mutations can lead to loss of immune control of HIV replication. Theoretically, escape from CD8(+) T cell recognition is less likely when multiple TCRs target individual MHC/peptide complexes, thereby increasing the chance that amino acid changes in the epitope could be tolerated. We studied the CD8(+) T cell response to six immunodominant epitopes in five HIV-infected subjects using a novel approach combining peptide stimulation, cell surface cytokine capture, flow cytometric sorting, anchored RT PCR, and real-time quantitative clonotypic TCR tracking. We found marked variability in the number of clonotypes targeting individual epitopes. One subject recognized a single epitope with six clonotypes, most of which were able to recognize and lyse cells expressing a major epitope variant that arose. Additionally, multiple clonotypes remained expanded during the course of infection, irrespective of epitope variant frequency. Thus, CD8(+) T cells comprising multiple TCR clonotypes may expand in vivo in response to individual epitopes, and may increase the ability of the response to recognize virus escape mutants. PMID- 11884483 TI - Murine sclerodermatous graft-versus-host disease, a model for human scleroderma: cutaneous cytokines, chemokines, and immune cell activation. AB - Murine sclerodermatous graft-vs-host disease (Scl GVHD) models human scleroderma, with prominent skin thickening, lung fibrosis, and up-regulation of cutaneous collagen mRNA. Fibrosis in Scl GVHD may be driven by infiltrating TGF-beta1 producing mononuclear cells. Here we characterize the origin and types of those cutaneous effector cells, the cytokine and chemokine environments, and the effects of anti-TGF-beta Ab on skin fibrosis, immune cell activation markers, and collagen and cytokine synthesis. Donor cells infiltrating skin in Scl GVHD increase significantly at early time points post-transplantation and are detectable by PCR analysis of Y-chromosome sequences when female mice are transplanted with male cells. Cutaneous monocyte/macrophages and T cells are the most numerous cells in Scl GVHD compared with syngeneic controls. These immune cells up-regulate activation markers (MHC class II I-A(d) molecules and class A scavenger receptors), suggesting Ag presentation by cutaneous macrophages in early fibrosing disease. Early elevated cutaneous mRNA expression of TGF-beta1, but not TGF-beta2 or TGF-beta3, and elevated C-C chemokines macrophage chemoattractant protein-1, macrophage inflammatory protein-1alpha, and RANTES precede subsequent skin and lung fibrosis. Therefore, TGF-beta1-producing donor mononuclear cells may be critical effector cells, and C-C chemokines may play important roles in the initiation of Scl GVHD. Abs to TGF-beta prevent Scl GVHD by effectively blocking the influx of monocyte/macrophages and T cells into skin and by abrogating up-regulation of TGF-beta1, thereby preventing new collagen synthesis. The Scl GVHD model is valuable for testing new interventions in early fibrosing diseases, and chemokines may be new potential targets in scleroderma. PMID- 11884485 TI - Increased severity of experimental allergic encephalomyelitis in lyn-/- mice in the absence of elevated proinflammatory cytokine response in the central nervous system. AB - lyn, a member of the src kinase family, is an important signaling molecule in B cells. lyn(-/-) mice display hyperactive B-1 cells and IgM hyperglobulinemia. The role of lyn on T cell function and development of Th1-mediated inflammatory disease is not known. Therefore, we examined the effect of disruption of the lyn gene on the development of experimental allergic encephalomyelitis (EAE), a well established Th1-mediated autoimmune disease. Following immunization with myelin oligodendrocyte protein (MOG) p35-55, lyn(-/-) mice had higher clinical and pathological severity scores of EAE when compared with wild type (WT). The increase in the severity of EAE in lyn(-/-) mice was not associated with a commensurate increase in the production of proinflammatory cytokines in the CNS. lyn(-/-) mice with EAE showed elevation in serum anti-IgM MOG Ab levels over that seen in WT mice, along with a modest increase in the mRNA levels of complement C5 and its receptor, C5aR, in the spinal cord. Transfer of serum from MOG-immunized lyn(-/-) mice worsened EAE in WT mice, suggesting a pathogenic role for anti-MOG IgM Abs in EAE. These observations underscore the potential role of lyn in regulation of Th1-mediated disease and the role of autoantibodies and complement in the development of EAE. PMID- 11884486 TI - Poststress motionlike artifacts caused by the use of a dual-head gamma camera for (201)Tl myocardial SPECT. AB - When performing (201)Tl myocardial SPECT using a dual-head gamma camera on patients after exercise stress, we have observed in some a sudden increase in the counting rate between the 16th and 17th images. This increase provoked motionlike artifacts, which increased the number of false-positive findings. The aim of our study was to determine possible causes for this leap in activity. METHODS: We performed myocardial SPECT using a dual-head gamma camera on 110 patients after exercise stress: in 38 patients approximately 5 min after injection (group 1), in 43 patients approximately 14 min after injection (group 2), and in 29 patients twice, at approximately 5 and 20 min after injection (group 3). We also performed dynamic data acquisition for 10 min on 18 patients after exercise stress. We compared activity in the heart region in image series obtained after exercise stress and at rest. RESULTS: Daily quality control tests eliminated the possibility of any malfunctions of the gamma camera. Careful image analysis showed no visible patient motion. Our results showed that upward creep of the heart could not be a cause of the described phenomenon. After exercise stress, a > or = 5% activity leap in the heart region on the 16th and 17th frames was more frequent in group 1 than in group 2. Two consecutive acquisitions after exercise stress showed that the leap was >5% in 24 patients (83%) and 12 patients (41%) at the first and second acquisitions, respectively (group 3). In all patients, the leap was <5% at rest. Dynamic studies showed that the activity in the heart region steadily decreased in all patients after exercise stress. We suggest that decreasing (201)Tl concentrations in myocardium or blood could be a major reason for the described artifacts. CONCLUSION: We proposed that the pharmacokinetics of (201)Tl-chloride be evaluated within a short time after injection in humans after exercise stress. Now, in our department, we have begun acquisition approximately 12 min after (201)Tl administration, and the above-mentioned phenomenon has not appeared. However, to avoid the artifacts caused by early redistribution of (201)Tl, acquisition must not begin too late. PMID- 11884487 TI - The PET radioligand [carbonyl-(11)C]desmethyl-WAY-100635 binds to 5-HT(1A) receptors and provides a higher radioactive signal than [carbonyl-(11)C]WAY 100635 in the human brain. AB - 5-Hydroxytryptamine (serotonin)-1A (5-HT(1A)) receptors are of key interest in research on the pathophysiology and treatment of psychiatric disorders. The PET radioligand [carbonyl-(11)C]WAY-100635 ((11)C-WAY), where WAY-100635 is (3)H-(N (2-(1-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridyl) cyclohexane carboxamide, is commonly used for quantitation of 5-HT(1A) receptors in the human brain. The aim of this PET study was to compare (11)C-WAY with the putative metabolite and selective radioligand [carbonyl-(11)C]desmethyl-WAY-100635 ((11)C DWAY). METHODS: A PET examination was performed on each of 5 healthy male volunteers after intravenous injection of (11)C-WAY and (11)C-DWAY on separate occasions. Radioactive metabolites in plasma were determined with high performance liquid chromatography. The plasma metabolite--corrected input function was used in a kinetic compartment analysis. The simplified reference tissue model and peak equilibrium method, using the cerebellum as reference region, was applied for comparison of data. RESULTS: For both radioligands, the highest radioactivity was observed in the neocortex and the raphe nuclei, whereas radioactivity was low in the cerebellum. The regional binding potentials were similar for the 2 radioligands. The brain uptake was more than 2-fold higher for (11)C-DWAY than for (11)C-WAY, in part because of higher delivery (first-order rate constant K(1), 0.38 vs. 0.16). The time--activity curves were well described by a 3-compartment model for all regions, whereas uptake in the cerebellum could not be described by a 2-compartment model, supporting the existence of kinetically distinguishable nonspecific binding in the cerebellum or radioactive metabolites in the brain for both radioligands. Both radioligands were rapidly metabolized, and <10% of the radioactivity in plasma represented unchanged (11)C WAY or (11)C-DWAY at 10 min after injection. The metabolic pattern was similar for both radioligands, with the formation of radiolabeled cyclohexanecarboxylic acid and more polar components. For (11)C-WAY, small amounts of an additional labeled metabolite comigrated with reference desmethyl-WAY-100635. CONCLUSION: The advantages of (11)C-DWAY over (11)C-WAY for research on central 5-HT(1A) receptors is supported by a significantly higher radioactivity signal at equipotent doses, providing improved imaging statistics and advantages in biomathematic modeling and the preclusion of (11)C-DWAY as a metabolite interfering with PET measurements. PMID- 11884488 TI - Longitudinal evaluation of both morphologic and functional changes in the same individuals with Alzheimer's disease. AB - Morphologic and functional imaging studies have not always given concordant results about brain areas showing atrophic changes and reduced flow or metabolism in Alzheimer's disease (AD). The aim of this study was to determine the initial abnormality and the longitudinal changes in both morphologic and functional measurements for the same individuals with AD. METHODS: We investigated 15 patients with mild AD and 25 age-matched healthy volunteers. The AD patients underwent both MRI and SPECT 3 times at intervals of approximately 1 y. The gray matter volume, as segmented from MRI, and the regional cerebral blood flow (rCBF), as measured by SPECT, of AD patients were compared with those of healthy volunteers using statistical parametric mapping, which is a voxel-based analysis in stereotactic space. RESULTS: Considerable discordance between areas of regional atrophy and areas of decreased rCBF was observed. The medial temporal areas showed a faster and more extensive reduction of gray matter volume than of rCBF. In comparison with the value at the baseline study, rCBF in the posterior cingulate gyrus and precuneus and the associative parietal cortex was extensively decreased. In contrast, the extent of significant decrease in this area continued to be much narrower for gray matter volume than for rCBF, even in the follow-up studies. Frontal areas, including the anterior cingulate gyrus and the orbitofrontal areas, showed a progressive reduction in both rCBF and gray matter volume. The reduction in rCBF was in a more posterior part of the associative temporal cortex than was the reduction in gray matter volume. CONCLUSION: These results indicate a distinct discordance between morphologic and functional changes in a longitudinal study of AD. Functional changes may be caused partly by remote effects from the morphologically involved areas with decreased connectivity and partly by a compensatory response by neuronal plasticity. PMID- 11884489 TI - Quantitation of liver and spleen uptake of (99m)Tc-phytate colloid using SPECT: detection of liver cirrhosis. AB - The purpose of this study was to assess the diagnostic performance of quantitative SPECT of (99m)Tc-phytate colloid in detecting liver cirrhosis and to assess the correlation between the SPECT results and the severity of disease. METHODS: Quantitative SPECT was performed on 60 patients (38 men, 22 women; mean age, 62.4 y) with liver cirrhosis and 36 control patients (21 men, 15 women; mean age, 58.7 y) without liver cirrhosis, and the results for the 2 groups were compared. Correlation with Child--Pugh classification and receiver operating characteristic methodology was used to analyze the results. RESULTS: Cirrhotic livers had a lower total uptake than did control livers (35.6% plus minus 13.5% vs. 61.6% +/- 10.2%, P < 0.0001). This reduced uptake was associated with a significantly reduced percentage injected dose per cubic centimeter (%ID/cm(3)) (0.024 +/- 0.01 vs. 0.044 +/- 0.01, P < 0.0001). The volume, 1,467 +/- 348 cm(3), was similar to that of control livers (1,487 +/- 397 cm(3), P = 0.80). Total uptake in the spleen was significantly greater in patients with cirrhosis than in control patients (24.9% +/- 12% vs. 7.6% +/- 3.2%, P < 0.0001) because of an increased volume (833 +/- 460 cm(3) vs. 239 +/- 90 cm(3), P < 0.0001). The %ID/cm(3) of spleen tissue was 0.033 +/- 0.01, which was similar to the value in control patients (0.032 +/- 0.01, P = 0.88). Spleen volume showed the best performance in detecting liver cirrhosis, with a mean area under the curve of 0.97 (95% CI = 0.91--0.99). The severity of liver disease correlated better with total liver uptake (r = -0.68, 95% CI = -0.80 to -0.52, P < 0.0001). CONCLUSION: Individual SPECT quantitation of (99m)Tc-phytate colloid uptake in the liver and spleen could be used as a noninvasive method to separate normal from cirrhotic livers and to evaluate the severity of disease. PMID- 11884490 TI - Quantification of lymphatic function for investigation of lymphedema: depot clearance and rate of appearance of soluble macromolecules in blood. AB - The object of this study was to develop a new technique for the quantitative measurement of lymphatic function. The rate of clearance of radiolabeled protein from a subcutaneous depot is supplemented by measurement of the appearance of the protein in venous blood. This initial study was performed on normal arms, with a view to subsequent clinical application such as in the investigation of women with breast cancer--related lymphedema (BCRL). METHODS: Fourteen healthy volunteers (12 women, 2 men) and 8 women awaiting surgery for breast cancer were recruited for the study. Each received subcutaneous depot injection of protein solution in the second dorsal web space of each hand, labeled with (111)In on one side and with (99m)Tc on the other side. Human serum albumin (HSA) was the protein used in the first 8 subjects and human polyclonal immunoglobulin G (HIgG) was used thereafter. The activity at each depot was measured at regular intervals using a collimated sodium iodide scintillation detector, and the activity in venous blood sampled from both arms was measured in an automatic sample counter. RESULTS: (99m)Tc-HSA cleared from the depot consistently faster than (111)In-HSA (P = 0.001). The proportions of radionuclide remaining bound to protein in venous blood were higher for (99m)Tc than for (111)In. HIgG displayed improved labeling stability for both nuclides, reflected in equal rates of clearance. Blood activity rose steadily after an early latent phase and for HIgG correlated strongly with the rate of clearance from the depot (P < 0.001). Marked variation between individuals was observed. CONCLUSION: A dual-isotope technique relies on identical behavior of the 2 radiopharmaceuticals used. This study shows that this is the case with respect to HIgG but not HSA. (99m)Tc-HSA cleared faster than (111)In-HSA and yet displayed better in vivo labeling stability. We conclude that (111)In dissociates from HSA in the depot but then becomes locally bound. Using HIgG, a close correlation was observed between the rates of clearance from the depot and the appearance in venous blood. This finding suggests that HIgG would be a suitable marker for subsequent dual-isotope studies on women with BCRL. PMID- 11884491 TI - Whole-body (18)F-FDG PET and conventional imaging for predicting outcome in previously treated breast cancer patients. AB - This study was conducted to determine the ability of (18)F-FDG PET and conventional imaging (CI) to predict the outcomes in breast cancer patients who have previously undergone primary treatment. METHODS: The study population consisted of 61 female patients (median age, 54 y; range, 32--91 y) who were reevaluated with (18)F-FDG PET and CI after treatment. The median interval between the last treatment and PET was 0.4 y (range, 0--16 y). PET was performed within 3 mo of CI (median interval, 25 d; range, 2--84 d). To determine the independent impact of PET on outcome, PET images were reinterpreted in a blind fashion. Availability of clinical information after PET scanning (21 plus minus 12 mo) was required for study inclusion. Study endpoints were clinical evidence of progression of disease or death. RESULTS: Of 61 patients, 19 (31.1%) had no clinical evidence and 38 (62.3%) had evidence of residual or recurrent disease by the end of follow-up. Four patients (6.6%) had died. The positive and negative predictive values (PPV and NPV, respectively) of PET were 93% and 84%, respectively. CI yielded a PPV of 85% and an NPV of 59%. The prognostic accuracy of single whole-body PET was superior to that of multiple procedures with CI (90% vs. 75%; P < 0.05). Kaplan--Meier estimates of disease-free survival in patients with negative PET findings compared with those with positive PET findings revealed a significant difference between the 2 curves (log-rank test = 0.001). Kaplan--Meier estimates of disease-free survival stratified by CI results showed a marginally significant difference between CI-positive and CI-negative patients (log-rank test = 0.04). CONCLUSION: FDG PET can be used to improve prediction of the clinical outcome of previously treated breast cancer patients relative to what is achievable through CI alone. PMID- 11884492 TI - Interobserver agreement on captopril renography for assessing renal vascular disease. AB - Captopril-stimulated renography is widely used to screen selected groups of hypertensive patients for renal vascular disease. Evaluation of the test is a complex task. Lack of interobserver agreement on the assessment and interpretation of renographic parameters may contribute to differences in sensitivity and specificity between studies. METHODS: Three experienced nuclear medicine physicians evaluated 658 renograms of 503 hypertensive patients suspected of having renal vascular disease from a large Dutch multicenter study (the Dutch Renal Artery Stenosis Intervention Cooperative [DRASTIC] study). Interobserver agreement on several renographic parameters was assessed by the kappa statistic and the intraclass correlation coefficient (ICC). RESULTS: The interobserver agreement on the time to excretion was high: The pooled ICC was 0.90. The pooled kappa was > or = 0.65 for the pattern of the time--activity curves, the visual aspect of the scintigraphic images (visible uptake and kidney size), and the judgment on the presence of renal artery stenosis. However, the interobserver agreement on cortical retention and pelvic retention by visual inspection of the images was rather low (pooled kappa = 0.46 and 0.52, respectively). Pelvic retention was found to complicate the interpretation of renography. CONCLUSION: Interobserver agreement on most of the renographic parameters was satisfactory, but the assessment of cortical retention was more difficult, in particular, in the presence of pelvic retention. Captopril renography should be interpreted with caution if pelvic retention is suspected. Interobserver variability offers one of several explanations for the differences in diagnostic test performance that are found between studies. PMID- 11884493 TI - Quantitative studies of bone in postmenopausal women using (18)F-fluoride and (99m)Tc-methylene diphosphonate. AB - Quantitative radionuclide studies of bone using the short-lived tracers (18)F fluoride and (99m)Tc-methylene diphosphonate (MDP) are an alternative method to biochemical markers of bone turnover for investigating the dynamic state of the skeleton. In this study we evaluated their use to quantify bone turnover in women receiving antiresorptive therapy compared with that of untreated control subjects. METHODS: The patients were 69 healthy postmenopausal women. Twenty-six women were receiving hormone replacement therapy (HRT) and 43 were untreated age matched control subjects. After bolus injection of (18)F-fluoride (1 MBq), (99m)Tc-MDP (1 MBq), (51)Cr-ethylenediaminetetraacetic acid (3 MBq), and (125)I labeled human serum albumin (0.25 MBq), multiple blood samples and urine collections were taken between 0 and 4 h. The clearance to bone mineral K(bone) was first evaluated using the area under the plasma concentration curve (AUC) on the assumption that the rate constant k(4) for the outflow of tracer from bone was negligibly small. AUC values of K(bone) were then compared with those found using a compartmental model method that allowed k(4) to be fitted as a free parameter. RESULTS: Using the AUC method the mean plus minus SD for K(bone) for the 2 tracers were: (18)F-fluoride, 61.8 plus minus 12.0 mL center dot min(-1) (HRT group) versus 67.2 plus minus 12.6 mL center dot min(-1) (control group) (P = 0.045); and (99m)Tc-MDP, 40.3 plus minus 8.2 mL center dot min(-1) (HRT group) versus 44.2 plus minus 7.6 mL center dot min(-1) (control group) (P = 0.024). Values for the 2 tracers in individual patients were moderately well correlated (r = 0.76; P < 0.001). Using the compartmental model method, k(4) for (18)F fluoride was shown to lie in the range 0--0.0025 min(-1) with a best-fit value of 0.0018 min(-1). Values of K(bone) determined using k(4) = 0.0018 min(-1) were highly correlated with the AUC values (r = 0.989; SEE = 2.05 mL center dot min( 1)) with numeric values that were larger by a factor of 1.53. Analysis of the (99m)Tc-MDP data was more difficult because of uncertainties in protein binding in the extracellular fluid compartment space. The best fit for k(4) was in the range 0.0010--0.0014 min(-1) with values of K(bone) similar to those found using the AUC method. CONCLUSION: Values of K(bone) determined using the AUC method were able to differentiate between HRT-treated women and postmenopausal women who were not treated and were highly correlated with those determined using a compartmental model method with nonzero values of k(4). PMID- 11884494 TI - Heterotopic ossification. AB - Heterotopic ossification (HO) is the presence of bone in soft tissue where bone normally does not exist. The acquired form of HO most frequently is seen with either musculoskeletal trauma, spinal cord injury, or central nervous system injury. For example, patients who have recently undergone total hip arthroplasty or have paraplegia after spinal cord injury are at risk for HO. The fever, swelling, erythema, and occasional joint tenderness seen in early HO can be difficult to distinguish from cellulitis, osteomyelitis, or thrombophlebitis. Bone scanning and other imaging tests frequently are used to distinguish between these diagnostic possibilities. As treatment or prophylaxis for HO, either a nonsteroidal antiinflammatory drug (such as indomethacin), a diphosphonate (such as ethane-1-hydroxy-1,1-diphosphate), or local radiation therapy is recommended. Before therapy begins, bone scanning may be requested to confirm the diagnosis of HO. In addition, surgical resection of HO is used to preserve joint mobility; however, HO is likely to recur and possibly progress if resection is undertaken before the lesion has become mature. With a view toward avoiding recurrent HO and other operative complications, serial quantitative bone scans are used as an aid to time surgical intervention. PMID- 11884495 TI - A practical methodology for patient release after tositumomab and (131)I tositumomab therapy. AB - A methodology was developed determining patient releasability after radioimmunotherapy with tositumomab and (131)I-tositumomab for the treatment of non-Hodgkin's lymphoma. METHODS: Dosimetry data were obtained and analyzed after 157 administrations of (131)I-tositumomab to 139 patients with relapsed or refractory non-Hodgkin's lymphoma. Tositumomab and (131)I-tositumomab therapy included dosimetric (low activity) and therapeutic (high activity) administrations. For each patient, the total-body residence time was calculated after the dosimetric administration from total-body counts obtained over 6 or 7 d and was then used to determine the appropriate therapeutic activity to deliver a specific total-body radiation dose. Patient dose rates at 1 m were measured immediately after the therapeutic infusion. Patient-specific calculations based on the measured total-body residence time and dose rate for (131)I-tositumomab were derived to determine the patient's maximum releasable dose rate at 1 m, estimated radiation dose to maximally exposed individuals, and the amount of time necessary to avoid close contact with others. RESULTS: The mean administered activity (+/-SD), determined by dosimetry studies for each patient before therapy, was 3,108 +/- 1,073 MBq (84 +/- 29 mCi) (range, 1,221 +/- 5,957 MBq [33- 161 mCi]). Immediately after treatment, the mean measured dose rate (+/- SD) at 1 m was 0.109 +/- 0.032 mSv/h (10.9 +/- 3.2 mrem/h; range, 0.04--0.24 mSv/h [4--24 mrem/h]). The measured dose rates were 60% (range, 37%--90%; P < 0.0001) of the theoretic dose rates from a point source in air predicted using the dose equivalent rate per unit activity of (131)I (5.95 x 10(-5) mSv/MBq h [0.22 mrem/mCi h] at 1 m). The mean estimated radiation dose to the maximally exposed individual was 3.06 mSv (306 mrem) (range, 1.95--4.96 mSv [195--496 mrem]). On the basis of current regulatory patient-release criteria, all (131)I-tositumomab- treated patients were determined to be releasable by comparing the dose rate at 1 m with a predetermined maximum releasable dose rate. Detailed instructions were provided to limit family members' exposure. CONCLUSION: A methodology has been developed for the release of patients administered radioactive materials based on the new Nuclear Regulatory Commission regulations. This approach uses a patient specific dose calculation based on the measured total-body residence time and dose rate. This analysis shows the feasibility of outpatient radioimmunotherapy with tositumomab and (131)I-tositumomab. PMID- 11884496 TI - Definitive improvement in the approach to the treated patient as a radioactive source. PMID- 11884497 TI - Relative uptake, metabolism, and beta-receptor binding of (1R,2S)-4-(18)F fluorometaraminol and (123)I-MIBG in normotensive and spontaneously hypertensive rats. AB - The objective of the study was to compare relative uptake, metabolism, and beta receptor affinity of the new positron-emitting uptake-1 tracer (1R,2S)-4-(18)F fluorometaraminol (4-FM) with those of the SPECT pharmaceutical meta-(123)I iodobenzylguanidine (MIBG) in Wistar Kyoto (WKY) rats and spontaneously hypertensive (SHR) rats. METHODS: No-carrier-added 4-(18)F-FM was applied to SHR and WKY rats in vivo and to retrogradely perfused hearts in vitro. Cardiac and extracardiac distribution was assessed, and metabolite formation was determined by thin-layer chromatography. The in vivo experiments were repeated with no carrier-added (123)I-MIBG. By means of autoradiography, the beta-receptor affinity of 4-FM was compared with that of MIBG and propranolol (10 micromol/L) through displacement of (125)I-iodocyanopindolol (1.5 pmol/L) in slices of heart and spleen. RESULTS: Cardiomyopathic hearts showed heterogeneous 4-(18)F-FM uptake with gradients up to 3.6 in vivo and in vitro between different regions of the heart. Control hearts showed such gradients in 4-(18)F-FM uptake only in vitro. (123)I-MIBG exhibited a less heterogeneous in vivo distribution in SHR hearts. Extracardiac differences between WKY and SHR were found for uptake of 4 (18)F-FM in the spleen (63.3% plus minus 4% vs. 38.8% plus minus 5.7% of cardiac activity) and for renal uptake of (123)I-MIBG (373% plus minus 27% vs. 81.4% plus minus 17% of cardiac activity). Metabolites of 4-(18)F-FM were found only in the liver and those of (123)I-MIBG were found in the liver and kidney with a nearly equal relative fraction in both types of animals of about 20%, 60%, and 30%, respectively. 4-FM suppressed cardiac-specific beta-receptor binding of (125)I iodocyanopindolol in heart and spleen of both types of animals significantly, whereas MIBG had almost no effect. CONCLUSION: The more heterogeneous cardiac distribution of 4-(18)F-FM suggests that it reflects alterations in uptake-1 better than (123)I-MIBG in addition to the possibility of quantification and higher spatial resolution by PET compared with SPECT. Altered biotransformation in cardiomyopathic diseases may also impair the evaluation of (123)I-MIBG-SPECT data. The beta-receptor binding of 4-(18)F-FM must be further elucidated. PMID- 11884498 TI - Preclinical evaluation of a new, stabilized neurotensin(8--13) pseudopeptide radiolabeled with (99m)tc. AB - The rapid degradation of neurotensin (NT) limits its clinical use in cancer imaging and therapy. Thus, a new NT(8--13) pseudopeptide, NT-VIII, was synthesized. Some changes were introduced in the sequence of NT(8--13) to stabilize the molecule against enzymatic degradation: Arg(8) was N-methylated, and Lys and Tle replaced Arg(9) and Ile(12), respectively. Finally, (NalphaHis)Ac was coupled to the N-terminus for (99m)Tc(CO)(3) labeling. This peptide was characterized both in vitro and in vivo. METHODS: The new analog was labeled with (99m)Tc(CO)(3). Its metabolic stability was analyzed both in human plasma and in HT-29 cells. Binding properties, receptor downregulation, and internalization were tested with HT-29 cells. Biodistribution was evaluated in nude mice with HT 29 xenografts. RESULTS: (99m)Tc(CO)(3)NT-VIII showed a high stability in plasma, where most of the peptide remained intact after 24 h of incubation at 37 degreesC. However, the degradation in HT-29 cells was more rapid (46% of intact (99m)Tc(CO)(3)NT-VIII after 24 h at 37 degreesC). Binding to NT1 receptors (NTR1) was saturable and specific. Scatchard analysis showed a high affinity for (99m)Tc(CO)(3)NT-VIII, with a dissociation constant similar to (125)I-NT (1.8 vs. 1.6 nmol/L). After interacting with NTR1, (99m)Tc(CO)(3)NT-VIII was rapidly internalized, with more than 90% internalized after 30 min. It also distributed and cleared rapidly in nude mice bearing HT-29 xenografts. The highest rates of accumulation were found in kidney and tumor at all time points tested. Tumor uptake was highly specific because it could be blocked by coinjection with a high dose of (NalphaHis)Ac-NT(8--13). Tumors were clearly visualized in scintigraphy images. CONCLUSION: The changes that were introduced stabilized the molecule against enzymatic degradation without affecting binding properties. Moreover, the increase in stability enhanced tumor uptake, making this derivative a promising candidate for clinical use. PMID- 11884499 TI - Tumor pretargeting in mice using (99m)Tc-labeled morpholino, a DNA analog. AB - Over the past several years, investigators in this laboratory and elsewhere have been studying tumor localization by pretargeting with streptavidin and biotin or with avidin and biotin. Despite encouraging results, difficulties related to endogenous biotin and the immunogenicities of streptavidin and avidin have made a search for alternative strategies sensible. Recently, we have considered the use of DNAs and peptide nucleic acids for this purpose because oligomers can have hybridization affinities equivalent to that of biotin for streptavidin or avidin without the associated difficulties. We now report on the use of a morpholino (MORF), another commercially available synthetic oligomer, for pretargeting applications. MORFs support the nitrogenous bases by nonionic phosphorodiamidate linkages and, besides being nuclease resistant, can display good water solubility. METHODS: An 18mer MORF and its 18mer complementary MORF (cMORF) were obtained with a primary amine through a 3-member alkyl linker on the 3' equivalent end. An anti--carcinoembryonic antigen IgG antibody (MN14) was conjugated with MORF, whereas cMORF was conjugated with N-hydroxysuccinimide mercaptoacetyltriglycine (MAG3) to permit radiolabeling with (99m)Tc. The biodistribution of labeled cMORF was first evaluated in normal CD-1 mice. Subsequently, nude mice bearing LS174T tumors received 50 microg conjugated antibody 48 h before the administration of 1.0 microg (7.4 MBq) (99m)Tc-MAG3 cMORF. Control animals received the labeled cMORF without prior administration of the antibody. A clearing step was not used. RESULTS: Biodistributions in normal mice showed that (99m)Tc-MAG3-cMORF was excreted rapidly through the kidneys, with only 7 percentage injected dose (%ID) remaining within the whole body (excluding urine) at 3 h. In tumor-bearing mice at 24 h, only 11 %ID of the radioactivity remained in the whole body of study animals, and of this amount, 2 %ID/g was in tumor tissue. The sites with the highest %ID were the kidneys, at 4 %ID/g, and the blood, at 0.5 %ID/g; all other organs had <1 %ID/g. At the same time, values for the control animals were 5 %ID (whole body), 0.05 %ID/g (tumor), and 3 %ID (kidneys). All images reflected high uptake in the tumors and low uptake in the normal tissues of the study mice. CONCLUSION: Pretargeting using MORFs was effective in a mouse tumor model. PMID- 11884500 TI - In vivo imaging of human colon cancer xenografts in immunodeficient mice using a guanylyl cyclase C--specific ligand. AB - Guanylyl cyclase C (GC-C) is a transmembrane receptor expressed by human intestinal cells and primary and metastatic colorectal adenocarcinomas but not by extraintestinal tissues or tumors. The Escherichia coli heat-stable enterotoxin analog, STa (5--18), is a 14--amino acid peptide that selectively binds to the extracellular domain of GC-C with subnanomolar affinity. This study examined the utility of a radiolabeled conjugate of STa (5--18) to selectively target and image extraintestinal human colon cancer xenografts in vivo in nude mice. METHODS: The STa conjugate, ethoxyethyl-mercaptoacetamidoadipoylglycylglycine-STa (5--18) (NC100586), was synthesized and labeled with (99m)Tc to produce (99m)Tc NC100586. This compound was intravenously administered to nude mice bearing human colon cancer xenografts, and specific targeting was evaluated by biodistribution and gamma camera imaging. RESULTS: In CD-1 nude mice, biodistribution and scintigraphic imaging analyses showed selective uptake of (99m)Tc-NC100586 into human colon cancer xenografts that express GC-C but not into normal tissues that do not express GC-C. Similarly, (99m)Tc-NC100586 injected intravenously into CD-1 nude mice with human colon cancer hepatic metastases selectively accumulated in those metastases, and about 5-mm foci of tumor cells were visualized after ex vivo imaging of excised livers. Accumulation of (99m)Tc-NC100586 in human colon cancer xenografts reflected binding to GC-C because (99m)Tc-NC100588, an inactive analog that does not bind to GC-C, did not selectively accumulate in cancer xenografts compared with normal tissues. Also, coadministration of excess unlabeled STa (5--18) prevented accumulation of (99m)Tc-NC100586 in human colon cancer xenografts. Furthermore, (99m)Tc-NC100586 did not selectively accumulate in Lewis lung tumor xenografts, which do not express GC-C. CONCLUSION: This study showed that intravenously administered STa (5--18) selectively recognizes and binds to GC-C expressed by human colon cancer cells in vivo. Also shown was the ability to exploit this selective interaction to target imaging agents to extraintestinal human colon tumors in nude mice. These results suggest the utility of STa and GC-C for the development of novel targeted imaging and therapeutic agents with high specificity for metastatic colorectal tumors in humans. PMID- 11884501 TI - Targeting of endothelin receptors for molecular imaging of atherosclerosis in rabbits. AB - We wanted to determine whether a previously described in vivo accumulation of a (99m)Tc-labeled endothelin derivative in atherosclerotic lesions is mediated by binding to endothelin receptors. Furthermore, the expression of endothelin receptors in atherosclerotic lesions of 2 different rabbit animal models for atherosclerosis was to be evaluated to determine whether endothelin receptors generally are a suitable target for atherosclerosis imaging. METHODS: Normal vessels from untreated New Zealand White rabbits (NZW), balloon-denuded aortas from cholesterol-fed NZW, and atherosclerotic aortas from Watanabe Heritable Hyperlipidemic rabbits (WHHL) were used either as cross sections (cryosections) for receptor binding studies or for superfusion with a medium containing (125)I labeled endothelin-1 or the (99m)Tc-labeled endothelin derivative. RESULTS: Cross sections of aortas from untreated NZW contained 45 +/- 11.10(6) endothelin A receptors per square millimeter and 55 +/-11.10(6).endothelin B receptors per square millimeter, cross sections of balloon-denuded aortas from cholesterol-fed NZW contained 106 +/- 16.10(6) endothelin A receptors per square millimeter and 27 +/- 16.10(6) endothelin B receptors per square millimeter, and cross sections of atherosclerotic aortas from WHHL contained 40 +/- 13.10(6) endothelin A receptors per square millimeter and 5 +/- 13.10(6) endothelin B receptors per square millimeter. Balloon-denuded aortas from cholesterol-fed NZW (366 +/- 132 amol.mm(-2), P < 0.001) and atherosclerotic aortas from WHHL (338 +/- 175 amol.mm(-2), P < 0.002) accumulated significantly more of the (99m)Tc-labeled endothelin derivative than did vessels from control animals (137 +/- 26 amol.mm( 2)). On the contrary, (125)I-labeled endothelin-1--bound receptor mediated to superfused aortas from untreated NZW (12 +/- 9 amol.mm(-2)) and to balloon denuded aortas from cholesterol-fed NZW (19 +/- 5 amol.mm(-2)) but not to aortas from WHHL. This lack of receptor-specific accumulation of (125)I-endothelin-1 in atherosclerotic areas of WHHL aortas, and this receptor-specific accumulation in atherosclerotic balloon-denuded NZW aortas that does not significantly increase in comparison with normal aortas of untreated NZW, cause failure of endothelins to detect atherosclerotic lesions. CONCLUSION: Although the density and the ratio of endothelin receptor subtypes change because of the development of atherosclerotic lesions in rabbit aortas, endothelin receptor targeting for imaging of atherosclerosis is not suitable. PMID- 11884502 TI - Noninvasive monitoring of somatostatin receptor type 2 chimeric gene transfer. AB - Noninvasive monitoring of gene transfer will benefit basic research and patient care. Most gene-transfer imaging systems do not directly detect the gene of interest, and most do not exploit radiopharmaceuticals that have Food and Drug Administration approval for total-body use. (111)In-Octreotide is used clinically to locate tumors overexpressing primarily somatostatin receptor type 2 (SSTR2). We report the in vitro and in vivo detection of SSTR2 chimeric gene transfer with this radiopharmaceutical. METHODS: Full-length SSTR2A was ligated into a vector downstream of a 5' Igkappa leader sequence and the hemagglutinin A (HA) sequence. The vector plus insert was then introduced into HT1080 cells. Igkappa and HA domain functions were confirmed by immunologic methods. Receptor binding was studied in transfected cells incubated with (111)In-octreotide with and without somatostatin-28. Mice bearing tumors produced by transfected cells were injected with (111)In-octreotide for biodistribution and imaging studies. RESULTS: Cell membrane localization by the amino-terminal Igkappa domain was confirmed by immunofluorescence. The HA domain was identified by enzyme-linked immunosorbent assay, immunofluorescence, and Western blotting analysis with anti-HA antibodies. (111)In-Octreotide detected the SSTR2 portion of the fusion protein in vitro (receptor-binding assay) and in vivo (biodistribution studies and gamma-camera imaging). In addition, in vitro studies using either the anti-HA antibody or (111)In-octreotide correlated with biodistribution and imaging studies when cell clones expressing different levels of the fusion protein were tested. This approach may be feasible clinically because we were able to discern chimeric gene transfer in tumor-bearing animals with (111)In-octreotide at doses similar to those already used in humans. CONCLUSION: With this method it may be possible to monitor transfer of a gene of interest directly and noninvasively. PMID- 11884503 TI - Cis-4-[(18)F]fluoro-L-proline PET imaging of pulmonary fibrosis in a rabbit model. AB - A fluorinated analog of proline amino acid, cis-4-[(18)F]fluoro-L-proline (FP), was tested for potential use in PET for detection and evaluation of pulmonary response to respirable crystalline silica. The purpose of the study was to determine whether PET imaging with FP is sensitive for detection of pulmonary fibrosis. METHODS: Experimental silicosis was produced in rabbits by airway instillation of 300 mg respirable silica in 0.9% sterile saline; control rabbits received only saline. After 1, 2, 4, or 5 mo, animals were injected with 37 MBq (1 mCi) FP, and imaged in sets of 2 to 3 in a PET scanner using a dynamic scanning protocol over a 3-h period. Each imaging set contained at least 1 control rabbit. FP uptake in each lung was scored from 0 to 5 (PET score) by consensus of 3 readers blinded to animals' exposure status. Animals were humanely killed 2 d after the last imaging, and tissue sections from each lung lobe were graded from 0 to 5 by histopathology examination (histopathology score) for severity and distribution of fibrosis. RESULTS: Silicotic animals had significantly higher (P < 0.05) PET scores at each time point than did control animals. Repeated-measures ANOVA showed significant differences in PET scores between silicotic and control animals for the total lung field, but there were no statistically significant time trends for either group. Presence of fibrosis (i.e., histopathology score > 1) showed a significant association with elevated PET score (i.e., PET score > 1) using Fisher's exact test (P < 0.05). PET scores also showed excellent predictive ability, as all animals (18/18) with fibrosis also had elevated PET scores, and 95% (18/19) of animals with PET scores > 1 showed evidence of fibrosis. Localization of activity to specific lung areas was less exact, perhaps due in part to the small animal size for the resolution of the clinical PET imager used. PET scores were elevated (>1) for 67% (10/15) of silicotic right lungs and 75% (12/16) of silicotic left lungs; fibrosis scores > 1 were measured in 91% (10/11) of right lungs with PET scores > 1, and in 92% (12/13) of such left lungs. CONCLUSION: The FP tracer provided sensitive and specific identification of silicotic animals in early stages of the disease. This suggests that FP PET imaging has the potential sensitivity to detect active fibrosis in silicosis and other lung diseases. Additional studies are needed to determine the specificity of the FP tracer for fibrosis versus inflammatory processes. PMID- 11884504 TI - Correlation of myocardial p-(123)I-iodophenylpentadecanoic acid retention with (18)F-FDG accumulation during experimental low-flow ischemia. AB - Myocardial ischemia is associated with reduced free fatty acid (FFA) beta oxidation and increased glucose utilization. This study evaluated the potential of dynamic SPECT imaging of a FFA analog, p-(123)I-iodophenylpentadecanoic acid (IPPA), for detection of ischemia and compares retention of IPPA with (18)F-FDG accumulation. METHODS: In a canine model of regional low-flow ischemia (n = 9), serial IPPA SPECT images (2 min per image) were acquired over 52--90 min. In a subset of dogs (n = 6), (18)F-FDG was injected after completing SPECT imaging and allowed to accumulate for 40 min before killing the animals. Flow was assessed with radiolabeled microspheres. Myocardial metabolism was evaluated independently by selective coronary arterial and venous sampling. RESULTS: Serial IPPA SPECT images showed an initial defect in the ischemic region (0.70% plus minus 0.03% ischemic-to-nonischemic ratio), which normalized within 48 min because of the slower IPPA clearance from the ischemic region (t(1/2) = 54.2 plus minus 3.3 min) relative to the nonischemic region (t(1/2) = 36.7 plus minus 5.6 min) (P < 0.05). Delayed myocardial IPPA and (18)F-FDG activities were correlated (r = 0.70; n = 576 segments), and both were maximally increased in segments with a moderate flow reduction (IPPA, 151% of nonischemic; (18)F-FDG, 450% of nonischemic; P < 0.05). CONCLUSION: Serial SPECT imaging showed delayed myocardial clearance of IPPA in ischemic regions with moderate flow reduction, which lead to increased late myocardial retention of IPPA. Retention of IPPA correlated with (18)F-FDG accumulation, supporting the potential of IPPA as a noninvasive marker of ischemic myocardium. PMID- 11884505 TI - Optimum compensation method and filter cutoff frequency in myocardial SPECT: a human observer study. AB - Attenuation, photon scatter, and distance-dependent collimator-detector response are major degrading factors in myocardial SPECT images. The current study investigated whether compensation for these factors improves perfusion defect detectability, and compared the results for human observers with a previous study using a mathematical observer. METHODS: Four methods were investigated: attenuation compensation (AC); attenuation and detector response compensation; attenuation and scatter compensation; and attenuation, detector response, and scatter compensation (ADSC). For ADSC, 4 three-dimensional postreconstruction Butterworth filter cutoff frequencies were investigated for a pixel size of 0.62 cm: 0.12, 0.14, 0.16, and 0.22 pixel(-1). Five observers read images reconstructed using the 4 compensation methods. Receiver operating characteristics (ROC) analysis was used to determine the area under the ROC curve in each treatment studied. RESULTS: Reconstruction methods that incorporated scatter and detector response compensation had higher indices of detectability than AC alone. Over the range studied, a filter cutoff frequency of 0.14 pixel( 1) was optimal. A comparison of human observer results with an earlier channelized Hotelling observer study performed with the same images showed excellent agreement in trend and ranking of defect detectability. CONCLUSION: Compensation for detector response and scatter improves defect detectability compared with AC alone, although detectability may depend on phantom population choice and noise level. An optimal filter cutoff was found that is lower than what is typically used in a clinical setting. The channelized Hotelling observer is a good predictor of human observer performance and may reduce the need for tedious, time-consuming studies with human observers. PMID- 11884506 TI - Quantification of (18)F-FDG uptake in the liver using dynamic PET. PMID- 11884507 TI - A tabulated summary of the FDG PET literature. PMID- 11884508 TI - In (anonymous) support of anonymity. By Caveman. PMID- 11884509 TI - Notch: a membrane-bound transcription factor. PMID- 11884510 TI - The PX domain: a new phosphoinositide-binding module. AB - The PX domain, which until recently was an orphan domain, has emerged as the latest member of the phosphoinositide-binding module superfamily. Structural studies have revealed that it has a novel fold and identified key residues that interact with the bound phosphoinositide, enabling some prediction of phosphoinositide-binding specificity. Specificity for PtdIns(3)P appears to be the most common, and several proteins containing PX domains localise to PtdIns(3)P-rich endosomal and vacuolar structures through their PX domains: these include the yeast t-SNARE Vam7p, mammalian sorting nexins (involved in membrane trafficking events) and the Ser/Thr kinase CISK, which is implicated in cell survival. Additionally, phosphoinositide binding to the PX domains of p40(phox) and p47(phox) appears to play a critical role in the active assembly of the neutrophil oxidase complex. PMID- 11884511 TI - Gene discovery by e-genetics: Drosophila odor and taste receptors. AB - A new algorithm that examines DNA databases for proteins that have a particular structure, as opposed to a particular sequence, represents a novel 'e-genetics' approach to gene discovery. The algorithm has successfully identified new G protein-coupled receptors, which have a characteristic seven-transmembrane-domain structure, from the Drosophila genome database. In particular, it has revealed novel families of odor receptors and taste receptors, which had long eluded identification by other means. The two new gene families, the Or and Gr genes, are expressed in neurons of olfactory and taste sensilla and are highly divergent from all other known G-protein-coupled receptor genes. Modification of the algorithm should allow identification of other classes of multitransmembrane domain protein. PMID- 11884512 TI - Cell-cycle-dependent localisation of Ulp1, a Schizosaccharomyces pombe Pmt3 (SUMO)-specific protease. AB - We report here on the characterisation of Ulp1, a component of the SUMO modification process in S. pombe. Recombinant S. pombe Ulp1 has de-sumoylating activity; it is involved in the processing of Pmt3 (S. pombe SUMO) and can, to a limited extent, remove Pmt3 from modified targets in S. pombe cell extracts. ulp1 is not essential for cell viability, but cells lacking the gene display severe cell and nuclear abnormalities. ulp1-null (ulp1.d) cells are sensitive to ultraviolet radiation in a manner similar to rad31.d and hus5.62, which have mutations in one subunit of the activator and the conjugator for the ubiquitin like protein SUMO respectively. However ulp1.d cells are less sensitive to ionising radiation and hydroxyurea (HU) than are rad31.d and hus5.62. ulp1-null cells are defective in processing precursor Pmt3 and display reduced levels of Pmt3 conjugates compared with wild-type cells. The slow growth phenotype of ulp1 null cells is not substantially rescued by over-expression of the mature form of Pmt3 (Pmt3-GG), suggesting that the de-conjugating activity of Ulp1 is required for normal cell cycle progression. During the S and G2 phases of the cell cycle the Ulp1 protein is localised to the nuclear periphery. However, during mitosis the pattern of staining alters, and during anaphase, Ulp1 is observed within the nucleus. Ulp1 localisation at the nuclear periphery is generally re-established by the time of septation (S phase). PMID- 11884513 TI - The Cdc42 and Rac1 GTPases are required for capillary lumen formation in three dimensional extracellular matrices. AB - Here we show a requirement for the Cdc42 and Rac1 GTPases in endothelial cell (EC) morphogenesis in three-dimensional extracellular matrices. Cdc42 and Rac1 specifically regulate EC intracellular vacuole and lumen formation in both collagen and fibrin matrices. Clostridium difficile toxin B (which blocks all three Rho GTPases) completely inhibited the ability of ECs to form both vacuoles and lumens, whereas C3 transferase, a selective inhibitor of Rho, did not. Expression of either dominant-negative (N17) or constitutively active (V12) Cdc42 using recombinant adenoviruses dramatically inhibited EC vacuole and lumen formation in both collagen and fibrin matrices. Both vacuole and lumen formation initiated in ECs expressing dominant-negative (N17) Rac1 but later collapsed, indicating a role for Rac1 during later stages of vessel development. Analysis of cultures using confocal microscopy revealed green fluorescent protein-V12Rac1, Rac1 wild-type and -Cdc42 wild-type chimeric proteins targeted to intracellular vacuole membranes during the lumen formation process. Also, expression of the verprolin-cofilin-acidic domain of N-WASP, a downstream Cdc42 effector, in ECs completely interfered with vacuole and lumen formation. These results collectively reveal a novel role for Cdc42 and Rac1 in the process of EC vacuole and lumen formation in three-dimensional extracellular matrices. PMID- 11884514 TI - Multi-parameter analysis of the kinetics of NF-kappaB signalling and transcription in single living cells. AB - Proteins of the NF-kappaB transcription factor family normally reside in the cytoplasm of cells in a complex with IkappaB inhibitor proteins. Stimulation with TNFalpha leads to proteosomal degradation of the IkappaB proteins and nuclear translocation of the NF-kappaB proteins. Expression of p65 and IkappaBalpha fused to fluorescent proteins was used to measure the dynamics of these processes in transfected HeLa cells. Simultaneous visualisation of p65-dsRed translocation and IkappaBalpha-EGFP degradation indicated that in the presence of dual fluorescent fusion protein expression, the half-time of IkappaBalpha-EGFP degradation was reduced and that of p65 translocation was significantly increased when compared with cells expressing the single fluorescent fusion proteins. These results suggest that the ratio of IkappaBalpha and p65 determine the kinetics of transcription factor translocation into the nucleus and indicate that the complex of p65 and IkappaBalpha is the true substrate for TNFalpha stimulation in mammalian cells. When cells were treated with the CRM-1-dependent nuclear export inhibitor, leptomycin B (LMB), there was nuclear accumulation of IkappaBalpha EGFP and p65-dsRed, with IkappaBalpha-EGFP accumulating more rapidly. No NF kappaB-dependent transcriptional activation was seen in response to LMB treatment. Following 1 hour treatment with LMB, significant IkappaBalpha-EGFP nuclear accumulation, but low levels of p65-dsRed nuclear accumulation, was observed. When these cells were stimulated with TNFalpha, degradation of IkappaBalpha-EGFP was observed in both the cytoplasm and nucleus. A normal transient transcription response was observed in the same cells using luminescence imaging of NF-kappaB-dependent transcription. These observations suggest that both normal activation and post-induction repression of NF-kappaB dependent transcription occur even when nuclear export of NF-kappaB is inhibited. The results provide functional evidence that other factors, such as modification of p65 by phosphorylation, or interaction with other proteins such as transcriptional co-activators/co-repressors, may critically modulate the kinetics of transcription through this signalling pathway. PMID- 11884515 TI - Imaging of procollagen transport reveals COPI-dependent cargo sorting during ER to-Golgi transport in mammalian cells. AB - We have examined the ER-to-Golgi transport of procollagen, which, when assembled in the lumen of the ER, is thought to be physically too large to fit in classically described 60-80 nm COPI- and COPII-coated transport vesicles. We found that procollagen exits the ER via COPII- coated ER exit sites and is transported to the Golgi along microtubules in defined transport complexes. These procollagen-containing transport complexes are, however, distinct from those containing other cargo proteins like ERGIC-53 and ts-045-G. Furthermore, they do not label for the COPI coat complex in contrast to those containing ts-045-G. Inhibition of COPII or COPI function before addition of ascorbate, which is required for the folding of procollagen, inhibits export of procollagen from the ER. Inactivation of COPI coat function after addition of ascorbate results in the localisation of procollagen to transport complexes that now also contain ERGIC-53 and are inhibited in their transport to the Golgi complex. These data reveal the existence of an early COPI-dependent, pre-Golgi cargo sorting step in mammalian cells. PMID- 11884516 TI - Association of the tetraspanin CD151 with the laminin-binding integrins alpha3beta1, alpha6beta1, alpha6beta4 and alpha7beta1 in cells in culture and in vivo. AB - CD151 is a cell surface protein that belongs to the tetraspanin superfamily. It forms complexes with the laminin-binding integrins alpha3beta1, alpha6beta1 and alpha6beta4 and is codistributed with these integrins in many tissues at sites of cell-matrix interactions. In this study we show that CD151 can also form stable complexes with the laminin-binding integrin alpha7beta1. The strength of this interaction is comparable to that between CD151 and alpha3beta1. Complexes of alpha3beta1, alpha6beta1 and alpha7beta1 with CD151 are equally well formed with all splice variants of the alpha3, alpha6 and alpha7 subunits, and complex formation is not affected by mutations that prevent the cleavage of the integrin alpha6 subunit. Like the expression of alpha3beta1 and alpha6beta1, expression of alpha7beta1 in K562 cells results in increased levels of CD151 at its surface. Two non-integrin laminin receptors, dystroglycan and the polypeptide on which the Lutheran blood group antigens are expressed, are also often colocalized with CD151, but no association with CD151-alpha3beta1 complexes was found with biochemical analysis. The anti-CD151 antibody TS151R detects an epitope at a site at which CD151 interacts with integrins, and therefore it cannot react with CD151 when it is bound to an integrin. Comparison of the straining patterns produced by TS151R with that by of an anti-CD151 antibody recognizing an epitope outside the binding site (P48) revealed that most tissues expressing one or more laminin binding integrins reacted with P48 but not with TS151R. However, smooth muscle cells that express alpha7beta1 and renal tubular epithelial cells that express alpha6beta1 were stained equally well by TS151R and P48. These results suggest that the interactions between CD151 and laminin-binding integrins are subject to cell-type-specific regulation. PMID- 11884517 TI - Mitochondrial oxidative stress and cell death in astrocytes--requirement for stored Ca2+ and sustained opening of the permeability transition pore. AB - The role of oxidative stress is established in a range of pathologies. As mitochondria are a major source of reactive oxygen species (ROS), we have developed a model in which an intramitochondrial photosensitising agent is used to explore the consequences of mitochondrial ROS generation for mitochondrial function and cell fate in primary cells. We have found that, in astrocytes, the interplay between mitochondrial ROS and ER sequestered Ca2+ increased the frequency of transient mitochondrial depolarisations and caused mitochondrial Ca2+ loading from ER stores. The depolarisations were attributable to opening of the mitochondrial permeability transition pore (mPTP). Initially, transient events were seen in individual mitochondria, but ultimately, the mitochondrial potential (Deltapsi(m)) collapsed completely and irreversibly in the whole population. Both ROS and ER Ca2+ were required to initiate these events, but neither alone was sufficient. Remarkably, the transient events alone appeared innocuous, and caused no increase in either apoptotic or necrotic cell death. By contrast, progression to complete collapse of Deltapsi(m) caused necrotic cell death. Thus increased mitochondrial ROS generation initiates a destructive cycle involving Ca2+ release from stores and mitochondrial Ca2+-loading, which further increases ROS production. The amplification of oxidative stress and Ca2+ loading culminates in opening of the mPTP and necrotic cell death in primary brain cells. PMID- 11884518 TI - Hepatocytes convert to a fibroblastoid phenotype through the cooperation of TGF beta1 and Ha-Ras: steps towards invasiveness. AB - In hepatocarcinogenesis, it is an open question whether transforming growth factor (TGF)-beta1 provides a tumor-suppressive or a tumor-promoting role. To address this question, we employed immortalized murine hepatocytes, which display a high degree of differentiation and, expectedly, arrest in the G1 phase under exposure to TGF-beta1. These hepatocytes maintain epithelial polarization upon expression of oncogenic Ha-Ras. However, Ras-transformed hepatocytes rapidly convert to a spindle-shaped, fibroblastoid morphology upon treatment with TGF beta1, which no longer inhibits proliferation. This epithelial to fibroblastoid conversion (EFC) is accompanied by disruption of intercellular contacts and remodeling of the cytoskeletal framework. Fibroblastoid derivatives form elongated branching cords in collagen gels and grow to severely vascularized tumors in vivo, indicating their increased malignancy and even invasive phenotype. Additionally, fibroblastoid cells secrete strongly enhanced levels of TGF-beta1, suggesting an autocrine regulation of TGF-beta signaling. Expression profiling further revealed that the loss of the adhesion component E-cadherin correlates with the upregulation of its transcriptional repressor Snail in fibroblastoid cells. Moreover, the phosphoinositide 3-OH (PI3) kinase pathway was required for the maintenance of EFC, as inhibition of PI3 kinase reverted fibroblastoid cells to an epithelial-like phenotype. Taken together, these data indicate a dual role of TGF-beta1 in hepatocytes: it induces proliferation arrest but provides a crucial function in promoting late malignant events in collaboration with activated Ha-Ras. PMID- 11884519 TI - Proprotein convertases are important mediators of the adipocyte differentiation of mouse 3T3-L1 cells. AB - Mouse 3T3-L1 cells are widely used to study adipocyte differentiation in vitro. When treated with insulin, dexamethasone and isobutylmethylxanthine these fibroblastic cells differentiate into round triglyceride-rich adipocytes. Because several proteins implicated in adipocyte differentiation (e.g. type 1 IGF receptors) are proteolytically activated by endoproteinases of the proprotein convertase family, we sought to determine whether these endoproteinases are crucial for adipose conversion. In this study, we show that expression of the proprotein convertases PACE4, PC7 and furin increases when 3T3-L1 cells are induced to differentiate into adipocytes. The differentiation was blocked in transfected cells expressing alpha1-antitrypsin Portland or in normal cells pre treated with the synthetic inhibitor decanoyl-RVKR-chloromethylketone. Both inhibitors are known to specifically inactivate proprotein convertases. The block was associated with impaired proteolytic activation of proIGF-1 receptor, absence of induction of the adipogenic transcriptional factor PPARgamma and marked reduction of the nuclear translocation of the C/EBPbeta factor. Taken together, these data constitute evidence that proprotein convertases are crucial mediators of adipogenesis. PMID- 11884520 TI - Chromosome associations in budding yeast caused by integrated tandemly repeated transgenes. AB - The binding of GFP-tagged tetracycline repressor (TetR) molecules to chromosomally integrated tetracycline operator (tetO) sequence repeats has been used as a system to study chromosome behaviour microscopically in vivo. We found that these integrated transgenes influence the architecture of yeast interphase nuclei, as chromosomal loci with tandem repeats of exogenous tetO sequences are frequently associated. These associations occur only if TetR molecules are present. tetO tandem repeats associate regardless of their chromosomal context. When they are present at a proximal and a distal chromosomal position, they perturb the normal polarized Rabl-arrangement of chromosome arms by recruiting chromosome ends to the centromeric pole of the nucleus. Associations are established at G1 and are reduced during S-phase and mitosis. This system may serve as a model for the role of DNA sequence-specific binding proteins in imposing nonrandom distribution of chromosomes within the nucleus. PMID- 11884521 TI - Dual labeling of the fibronectin matrix and actin cytoskeleton with green fluorescent protein variants. AB - We have prepared 3T3 cells doubly labeled to visualize simultaneously the extracellular fibronectin (FN) matrix and intracellular actin cytoskeleton in living cell cultures. We used FN-yellow fluorescent protein (FN-yfp) for the FN matrix, and the actin-binding domain of moesin fused to cyan fluorescent protein (cfp-Moe) to stain actin. Actin filament bundles were clearly seen in the protruding lamellae of the cells. FN matrix assembly appeared to be initiated as small spots of FN at the ends of actin filament bundles. The spots then elongated along the actin filament bundle toward the cell center to form FN fibrils. The end of the fibril towards the cell edge appeared immobile, and probably attached to the substrate, whereas the end toward the cell center frequently showed movements, suggesting attachment to the cell. Combining our data with the observations of Pankov et al. we suggest that fibrils grow by stretching this mobile end toward the cell center while adding new FN molecules at the end and along the entire length. When the cell culture was treated with cytochalasin to disrupt the actin cytoskeleton, some fibrils contracted substantially, suggesting that the segment attached primarily to the cell surface is stretched. PMID- 11884522 TI - Microvilli-like structures are associated with the internalization of virulent capsulated Neisseria meningitidis into vascular endothelial cells. AB - Bacterial pathogens are internalized into non-phagocytic cells either by a zipper mechanism involving a direct contact between a bacterial ligand and a cellular receptor or a trigger mechanism secondary to the formation of membrane ruffles. Here we show that internalization of capsulated Neisseria meningitidis within endothelial cells following type IV pilus-mediated adhesion is associated with the formation of cellular protrusions at the site of bacterial attachment. These protrusions, like microvilli, are highly enriched in ezrin and moesin, two members of the ERM (ezrin/radixin/moesin) family, whereas vinculin and paxillin are absent. ERM-binding transmembrane proteins, such as CD44, and cortical actin polymerization colocalized within these membrane protrusions. Expression of dominant-negative ezrin largely prevented cortical actin polymerization, thus confirming the role of this molecule in bacteria-induced cytoskeletal modifications. Moreover, using selective inhibitors and dominant-negative mutants of the Rho family GTPases, we show that bacteria-induced actin polymerization required the activation of both Rho and Cdc42 but not of Rac1. Whereas GTPase inhibition dramatically reduced actin polymerization at the site of bacterial attachment, ezrin recruitment was not affected, indicating that bacterial adhesion promotes ezrin recruitment independently of the activity of the Rho GTPases. Furthermore, GTPase inhibition largely reduced N. meningitidis entry into endothelial cells without affecting adhesion. We thus propose that following pilus-mediated adhesion, capsulated N. meningitidis recruit ERM-binding transmembrane proteins, as well as ezrin and moesin, and that both Rho and Cdc42 are critical for the subsequent cytoskeletal modifications responsible for the formation of microvilli-like cellular protrusions and bacterial internalization. PMID- 11884523 TI - A small RNA in testis and brain: implications for male germ cell development. AB - BC1 RNA, a small non-coding RNA polymerase III transcript, is selectively targeted to dendritic domains of a subset of neurons in the rodent nervous system. It has been implicated in the regulation of local protein synthesis in postsynaptic microdomains. The gene encoding BC1 RNA has been suggested to be a master gene for repetitive ID elements that are found interspersed throughout rodent genomes. A prerequisite for the generation of repetitive elements through retroposition and subsequent transmission in the germline is expression of the master gene RNA in germ cells. To test this hypothesis, we have investigated expression of BC1 RNA in murine male germ cells. We report that BC1 RNA is expressed at substantial levels in a subset of male germ cells. Results from cell fractionation experiments, developmental analysis, and northern and in situ hybridization showed that the RNA was expressed in pre-meiotic spermatogonia, with particularly high amounts in syncytial ensembles of cells that are primed for synchronous spermatogenic differentiation. BC1 RNA continued to be expressed in spermatocytes, but expression levels decreased during further spermatogenic development, and low or negligible amounts of BC1 RNA were identified in round and elongating spermatids. The combined data indicate that BC1 RNA operates in groups of interconnected germ cells, including spermatogonia, where it may function in the mediation of translational control. At the same time, the identification of BC1 RNA in germ cells provides essential support for the hypothesis that repetitive ID elements in rodent genomes arose from the BC1 RNA gene through retroposition. PMID- 11884524 TI - Three-dimensional reconstruction of cytoplasmic membrane networks in parietal cells. AB - There is general agreement that stimulation and consequent secretion of gastric parietal cells result in a great expansion of the apical canalicular membrane at the expense of an extensive intracellular network of membranes rich in the gastric proton pump (H,K-ATPase). However, there is ongoing controversy as to the precise nature of the intracellular membrane network, conventionally called tubulovesicles. At the heart of this controversy lies the question of whether tubulovesicles are a distinct membrane compartment or whether they are continuous with the apical plasma membrane. To address this controversy we used high pressure, rapid freezing techniques to fix non-stimulated (resting) rabbit gastric glands for electron microscopy. Ultra-thin (60-70 nm) serial sections were used for conventional TEM; 400-500 nm sections were used for tomography. Images were digitized and models constructed using Midas and Imod software (http://bio3d.colorado.edu ). Images were aligned and contours drawn on specific cellular structures. The contours from a stack of serial sections were arranged into objects and meshed into 3D structures. For resting parietal cells our findings are as follows: (1) The apical canaliculus is a microvilli-decorated, branching membrane network that extends into and throughout the parietal cell. This agrees well with a host of previous studies. (2) The plentiful mitochondria form an extensive reticular network throughout the cytoplasm. This has not previously been reported for the parietal cell, and the significance of this observation and the dynamics of the mitochondrial network remain unknown. (3) H,K ATPase-rich membranes do include membrane tubules and vesicles; however, the tubulovesicular compartment is chiefly comprised of small stacks of cisternae. Thus a designation of tubulocisternae seems appropriate; however, in the resting cell there are no continuities between the apical canaliculus and the tubulocisternae or between tubulocisternae. These data support the recruitment recycling model of parietal cell stimulation. PMID- 11884525 TI - Identification of septin-interacting proteins and characterization of the Smt3/SUMO-conjugation system in Drosophila. AB - The septins are a family of proteins involved in cytokinesis and other aspects of cell-cortex organization. In a two-hybrid screen designed to identify septin interacting proteins in Drosophila, we isolated several genes, including homologues (Dmuba2 and Dmubc9) of yeast UBA2 and UBC9. Yeast Uba2p and Ubc9p are involved in the activation and conjugation, respectively, of the ubiquitin-like protein Smt3p/SUMO, which becomes conjugated to a variety of proteins through this pathway. Uba2p functions together with a second protein, Aos1p. We also cloned and characterized the Drosophila homologues of AOS1 (Dmaos1) and SMT3 (Dmsmt3). Our biochemical data suggest that DmUba2/DmAos1 and DmUbc9 indeed act as activating and conjugating enzymes for DmSmt3, implying that this protein conjugation pathway is well conserved in Drosophila. Immunofluorescence studies showed that DmUba2 shuttles between the embryonic cortex and nuclei during the syncytial blastoderm stage. In older embryos, DmUba2 and DmSmt3 are both concentrated in the nuclei during interphase but dispersed throughout the cells during mitosis, with DmSmt3 also enriched on the chromosomes during mitosis. These data suggest that DmSmt3 could modify target proteins both inside and outside the nuclei. We did not observe any concentration of DmUba2 at sites where the septins are concentrated, and we could not detect DmSmt3 modification of the three Drosophila septins tested. However, we did observe DmSmt3 localization to the midbody during cytokinesis both in tissue-culture cells and in embryonic mitotic domains, suggesting that DmSmt3 modification of septins and/or other midzone proteins occurs during cytokinesis in Drosophila. PMID- 11884526 TI - CEACAM1 isoforms with different cytoplasmic domains show different localization, organization and adhesive properties in polarized epithelial cells. AB - CEACAM1 is a signaling cell adhesion molecule expressed in epithelia, vessel endothelia and leukocytes. It is expressed as two major isoforms with different cytoplasmic domains. CEACAM1 occurs both in cell-cell contact areas and on apical surfaces of polarized epithelial cells, but it is not known how the different isoforms are distributed in polarized cells or what the functions of CEACAM1 are in the apical surfaces. We investigated the localization and organization of the two CEACAM1 isoforms in transfected, polarized MDCK cells by confocal microscopy and differential surface labelling. CEACAM1-L was found on both the apical and the lateral surfaces, whereas CEACAM1-S appeared exclusively on the apical surfaces. Maintenance of the lateral localization of CEACAM1-L required homophilic binding between CEACAM1-L molecules on adjacent cells. Double labelling with anti-CEACAM1 antibodies directed against different epitopes indicated that apical CEACAM1-L occurred either in a homophilic adhesive state or in a free non-adhesive state. CEACAM1-S appeared almost exclusively in the homophilic adhesive state. These findings suggest that CEACAM1 mediates adhesive bonds between adjacent microvilli on the apical surfaces. PMID- 11884527 TI - Muscle regeneration by reconstitution with bone marrow or fetal liver cells from green fluorescent protein-gene transgenic mice. AB - The myogenic potential of bone marrow and fetal liver cells was examined using donor cells from green fluorescent protein (GFP)-gene transgenic mice transferred into chimeric mice. Lethally irradiated X-chromosome-linked muscular dystrophy (mdx) mice receiving bone marrow cells from the transgenic mice exhibited significant numbers of fluorescence(+) and dystrophin(+) muscle fibres. In order to compare the generating capacity of fetal liver cells with bone marrow cells in neonatal chimeras, these two cell types from the transgenic mice were injected into busulfantreated normal or mdx neonatal mice, and muscular generation in the chimeras was examined. Cardiotoxin-induced (or -uninduced, for mdx recipients) muscle regeneration in chimeras also produced fluorescence(+) muscle fibres. The muscle reconstitution efficiency of the bone marrow cells was almost equal to that of fetal liver cells. However, the myogenic cell frequency was higher in fetal livers than in bone marrow. Among the neonatal chimeras of normal recipients, several fibres expressed the fluorescence in the cardiotoxin untreated muscle. Moreover, fluorescence(+) mononuclear cells were observed beneath the basal lamina of the cardiotoxin-untreated muscle of chimeras, a position where satellite cells are localizing. It was also found that mononuclear fluorescence(+) and desmin(+) cells were observed in the explantation cultures of untreated muscles of neonatal chimeras. The fluorescence(+) muscle fibres were generated in the second recipient mice receiving muscle single cells from the cardiotoxin-untreated neonatal chimeras. The results suggest that both bone marrow and fetal liver cells may have the potential to differentiate into muscle satellite cells and participate in muscle regeneration after muscle damage as well as in physiological muscle generation. PMID- 11884528 TI - p65-NFkappaB synergizes with Notch to activate transcription by triggering cytoplasmic translocation of the nuclear receptor corepressor N-CoR. AB - Notch/RBP-Jkappa and nuclear factor-kappaB (NFkappaB) complexes are key mediators of the progression of many cellular events through the activation of specific target gene transcription. Independent observations have shown that activation of Notch-dependent transcription generally correlates with inhibition of differentiation. In contrast, activated NFkappaB complexes are required for progression of differentiation in several systems. Although some interactions between both pathways have been observed, the physiological significance of their connection is unclear. We have now demonstrated that the increase in p65-NFkappaB protein levels enhances Notch-mediated activation of the Hes1 promoter up to three-fold. This effect does not require NFkappaB transcriptional activity, and it is independent of the previously described interaction between Notch and p50 NFkappaB. Furthermore, we show that p65-NFkappaB can modulate subcellular localization of the transcriptional corepressor N-CoR, abrogating N-CoR mediated repression of the Hes1 promoter. In addition, p65-NFkappaB is able to upregulate not only the Hes1 but also other promoters containing SRE and AP-1 sites, which are repressed by N-CoR. Thus, we conclude that p65-NFkappaB can regulate gene expression by a general mechanism that involves cytoplasmic translocation of the transcriptional corepressor protein N-CoR. PMID- 11884529 TI - Dual regulation of telomerase activity through c-Myc-dependent inhibition and alternative splicing of hTERT. AB - Telomerase is believed to be induced upon proliferation and inhibited when cells differentiate. Thus, regulation of telomerase activity could be an important mechanism to limit growth of normal and cancer cells. Using transforming growth factor-beta 1 (TGF-beta1), which is known to control proliferation in epithelial cells, we now demonstrate that in the human HaCaT skin keratinocytes, TGF-beta1 downregulated c-Myc, and this blocked proliferation. This also caused a decrease in hTERT expression, which in turn inhibited telomerase activity. Overexpressing hTERT recovered telomerase activity but not proliferation, whereas constitutive expression of c-Myc recovered proliferation and hTERT expression. Nevertheless, telomerase remained inhibited, thus dissociating proliferation and telomerase activity. In addition, we show that TGF-beta1 inhibited telomerase activity despite ongoing hTERT transcription by inducing loss of the full-length hTERT transcript (mediating telomerase activity) and retaining high expression of the inactive beta variant. These changes in the splicing pattern reversed upon TGF beta1 removal, as did inhibition of telomerase activity, suggesting that alternative splicing may represent a novel mechanism of telomerase regulation by TGF-beta1. In addition, we show that destruction of tissue integrity (in a model for epidermal blistering) resulted in a rapid induction of the inactive beta variant, whereas tissue regeneration (formation of a stratified epithelium) correlated with a shift to the active full-length transcript, which is the dominant form in intact epidermis. Thus alternative splicing may not be restricted to TGF-beta1 but may add a more general mechanism of hTERT regulation in epidermal cells. PMID- 11884530 TI - Activation of protein kinase Ceta triggers cortical granule exocytosis in Xenopus oocytes. AB - Previous work has shown that phorbol esters or diacylglycerol trigger cortical granule exocytosis in Xenopus oocytes. We sought to identify the isoform(s) of protein kinase C (PKC) that mediate(s) this regulated secretory event. Because this process is initiated by lipid activators of PKC but is independent of calcium ions, we focused on the family of novel (calcium-independent) PKCs. Pharmacological investigations using Go6976 and Go6983 tended to exclude PKCdelta, epsilon and mu as secretory triggers. Subcellular fractionation and immunoblot data revealed that these oocytes expressed all five members of the novel PKC family, but it was only PKCeta that colocalized with cortical granules. Finally, expression of wild type or constitutively active forms of PKCdelta and eta strongly supported the conclusion that it is PKCeta that initiates cortical granule exocytosis in these cells. These observations represent an important step in identifying the mechanism of secretory triggering in this system. PMID- 11884531 TI - Rab5a GTPase regulates fusion between pathogen-containing phagosomes and cytoplasmic organelles in human neutrophils. AB - Biogenesis of phagolysosomes proceeds through a sequential series of interactions with endocytic organelles, a process known to be regulated by Rab and SNARE proteins. The molecular mechanisms underlying phagosome maturation in neutrophils are, however, not clearly understood. We investigated fusion between phagosomes containing the intracellular pathogen Mycobacterium tuberculosis versus the extracellular pathogen Staphylococcus aureus (designated MCP for mycobacteria containing phagosome and SCP for S. aureus-containing phagosome) and cytoplasmic compartments in human neutrophils. Western blot analysis of phagosomes isolated after internalisation revealed that lactoferrin (a constituent of secondary granules) and LAMP-1 were incorporated into both SCP and MCP, whereas hck (marker of azurophil granules) interacted solely with SCP. The subcellular distribution of the proteins Rab5a and syntaxin-4 suggested a role in docking of granules and/or endosomes to the target membrane in the neutrophil. We observed that during phagocytosis, Rab5a in GTP-bound form interacted with syntaxin-4 on the membrane of MCP and were retained for up to 90 minutes, whereas the complex was recruited to the SCP within 5 minutes but was selectively depleted from these vacuoles after 30 minutes of phagocytosis. Downregulation of Rab5a by antisense oligonucleotides efficiently reduced the synthesis of Rab5a, the binding of syntaxin-4 to MCP and SCP and the capacity for fusion exhibited by the pathogen containing phagosomes, but it had no effect on bacteria internalisation. These data indicate that the difference in granule fusion is correlated with a difference in the association of Rab5a and syntaxin-4 with the phagosomes. Intracellular pathogen-containing phagosomes retain Rab5a and syntaxin-4, whereas extracellular pathogen-containing phagosomes bind briefly to this complex. These results also identified Rab5a as a key regulator of phagolysosome maturation in human neutrophils. PMID- 11884532 TI - Cholesterol is important in control of EGF receptor kinase activity but EGF receptors are not concentrated in caveolae. AB - We have investigated the localization and function of the epidermal growth factor receptor (EGFR) in normal cells, in cholesterol-depleted cells and in cholesterol enriched cells. Using immunoelectron microscopy we find that the EGFR is randomly distributed at the plasma membrane and not enriched in caveolae. Binding of EGF at 4 degrees C does not change the localization of EGFR, and by immunoelectron microscopy we find that only small amounts of bound EGF localize to caveolae. However, upon patching of lipid rafts, we find that a significant amount of the EGFR is localized within rafts. Depletion of the plasma membrane cholesterol causes increased binding of EGF, increased dimerization of the EGFR, and hyperphosphorylation of the EGFR. Addition of cholesterol was found to reduce EGF binding and reduce EGF-induced EGFR activation. Our results suggest that the plasma membrane cholesterol content directly controls EGFR activation. PMID- 11884533 TI - Destiny of unspliced retroviral RNA: ribosome and/or virion? PMID- 11884534 TI - Novel swine virulence determinant in the left variable region of the African swine fever virus genome. AB - Previously we have shown that the African swine fever virus (ASFV) NL gene deletion mutant E70DeltaNL is attenuated in pigs. Our recent observations that NL gene deletion mutants of two additional pathogenic ASFV isolates, Malawi Lil-20/1 and Pr4, remained highly virulent in swine (100% mortality) suggested that these isolates encoded an additional virulence determinant(s) that was absent from E70. To map this putative virulence determinant, in vivo marker rescue experiments were performed by inoculating swine with infection-transfection lysates containing E70 NL deletion mutant virus (E70DeltaNL) and cosmid DNA clones from the Malawi NL gene deletion mutant (MalDeltaNL). A cosmid clone representing the left-hand 38-kb region (map units 0.05 to 0.26) of the MalDeltaNL genome was capable of restoring full virulence to E70DeltaNL. Southern blot analysis of recovered virulent viruses confirmed that they were recombinant E70DeltaNL genomes containing a 23- to 28-kb DNA fragment of the Malawi genome. These recombinants exhibited an unaltered MalDeltaNL disease and virulence phenotype when inoculated into swine. Additional in vivo marker rescue experiments identified a 20-kb fragment, encoding members of multigene families (MGF) 360 and 530, as being capable of fully restoring virulence to E70DeltaNL. Comparative nucleotide sequence analysis of the left variable region of the E70DeltaNL and Malawi Lil-20/1 genomes identified an 8-kb deletion in the E70DeltaNL isolate which resulted in the deletion and/or truncation of three MGF 360 genes and four MGF 530 genes. A recombinant MalDeltaNL deletion mutant lacking three members of each MGF gene family was constructed and evaluated for virulence in swine. The mutant virus replicated normally in macrophage cell culture but was avirulent in swine. Together, these results indicate that a region within the left variable region of the ASFV genome containing the MGF 360 and 530 genes represents a previously unrecognized virulence determinant for domestic swine. PMID- 11884535 TI - Efficient concerted integration by recombinant human immunodeficiency virus type 1 integrase without cellular or viral cofactors. AB - Replication of retroviruses requires integration of the linear viral DNA genome into the host chromosomes. Integration requires the viral integrase (IN), located in high-molecular-weight nucleoprotein complexes termed preintegration complexes (PIC). The PIC inserts the two viral DNA termini in a concerted manner into chromosomes in vivo as well as exogenous target DNA in vitro. We reconstituted nucleoprotein complexes capable of efficient concerted (full-site) integration using recombinant wild-type human immunodeficiency virus type I (HIV-1) IN with linear retrovirus-like donor DNA (480 bp). In addition, no cellular or viral protein cofactors are necessary for purified bacterial recombinant HIV-1 IN to mediate efficient full-site integration of two donor termini into supercoiled target DNA. At about 30 nM IN (20 min at 37 degrees C), approximately 15 and 8% of the input donor is incorporated into target DNA, producing half-site (insertion of one viral DNA end per target) and full-site integration products, respectively. Sequencing the donor-target junctions of full-site recombinants confirms that 5-bp host site duplications have occurred with a fidelity of about 70%, similar to the fidelity when using IN derived from nonionic detergent lysates of HIV-1 virions. A key factor allowing recombinant wild-type HIV-1 IN to mediate full-site integration appears to be the avoidance of high IN concentrations in its purification (about 125 microg/ml) and in the integration assay (<50 nM). The results show that recombinant HIV-1 IN may not be significantly defective for full-site integration. The findings further suggest that a high concentration or possibly aggregation of IN is detrimental to the assembly of correct nucleoprotein complexes for full-site integration. PMID- 11884537 TI - Capsid-specific cytotoxic T lymphocytes recognize three distinct H-2D(b) restricted regions of the BeAn strain of Theiler's virus and exhibit different cytokine profiles. AB - The role of virus-specific cytotoxic T lymphocytes (CTL) in Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease, a viral model for multiple sclerosis, is not yet clear. To investigate the specificity and function of CTL generated in response to TMEV infection, we generated a panel of overlapping 20-mer peptides encompassing the entire capsid and leader protein region of the BeAn strain of TMEV. Binding of these peptides to H-2K(b) and H 2D(b) class I molecules of resistant mice was assessed using RMA-S cells. Several peptides displayed significant binding to H-2K(b), H-2D(b), or both. However, infiltrating cytotoxic T cells in the central nervous system of virus-infected mice preferentially lysed target cells pulsed with VP2(111-130/121-140) or VP2(121-130), a previously defined CTL epitope shared by the DA strain of TMEV and other closely related cardioviruses. In addition, at a high effector-to target cell ratio, two additional peptides (VP2(161-180) and VP3(101-120)) sensitized target cells for cytolysis by infiltrating T cells or splenic T cells from virus-infected mice. The minimal epitopes within these peptides were defined as VP2(165-173) and VP3(110-120). Based on cytokine profiles, CTL specific for these subdominant epitopes are Tc2, in contrast to CTL for the immunodominant epitope, which are of the Tc1 type. Interestingly, CTL function towards both of these subdominant epitopes is restricted by the H-2D molecule, despite the fact that these epitopes bind both H-2K and H-2D molecules. This skewing toward an H 2D(b)-restricted response may confer resistance to TMEV-induced demyelinating disease, which is known to be associated with the H-2D genetic locus. PMID- 11884536 TI - A human immunodeficiency virus type 1 isolate from an infected person homozygous for CCR5Delta32 exhibits dual tropism by infecting macrophages and MT2 cells via CXCR4. AB - The mechanisms of human immunodeficiency virus (HIV) infection of a man (VH) homozygous for the CCR5Delta32 mutation were investigated, and coreceptors other than CCR5 used by HIV type 1 (HIV-1) isolated from this individual were identified. In contrast to previous reports, this individual's rate of disease progression was not accelerated. Homozygosity for CCR5Delta32 mutation was demonstrated by PCR and DNA sequencing (R. Biti et al., Nat. Med. 3:252-253, 1997). CCR5 surface expression was absent on T lymphocytes and macrophages. HIV was isolated by coculture with peripheral blood mononuclear cells (PBMCs) from siblings who were homozygous (VM) or wild type (WT) for the CCR5Delta32 mutation. The virus demonstrated dual tropism for infection of MT2 cell line and primary macrophages. Sequencing of the full HIV genome directly from the patient's PBMCs revealed 21 nucleotide insertions in the V1 region of gp120. The VH envelope sequence segregated apart from both the T-cell-line-adapted tropic strains NL4-3 and SF2 and M-tropic strain JRFL or YU2 by phylogenetic tree analysis. VH was shown to utilize predominantly CXCR4 for entry into T lymphocytes and macrophages by HOS.CD4 cell infection assay, direct envelope protein fusion, and inhibition by anti-CXCR4 monoclonal antibody (12G5), SDF-1, and AMD3100. Microsatellite mapping demonstrated the separate inheritance of CXCR4 by both homozygote brothers (VH and VM). Our study demonstrates the ability of certain strains of HIV to readily use CXCR4 for infection or entry into macrophages, which is highly relevant to the pathogenesis of late-stage disease and presumably also HIV transmission. PMID- 11884538 TI - Bipartite signal for genomic RNA dimerization in Moloney murine leukemia virus. AB - Retroviral virions each contain two identical genomic RNA strands that are stably but noncovalently joined in parallel near their 5' ends. For certain viruses, this dimerization has been shown to depend on a unique RNA stem-loop locus, called the dimer initiation site (DIS), that efficiently homodimerizes through a palindromic base sequence in its loop. Previous studies with Moloney murine leukemia virus (Mo-MuLV) identified two alternative DIS loci that can each independently support RNA dimerization in vitro but whose relative contributions are unknown. We now report that both of these loci contribute to the assembly of the Mo-MuLV dimer. Using targeted deletions, point mutagenesis, and antisense oligonucleotides, we found that each of the two stem-loops forms as predicted and contributes independently to dimerization in vitro through a mechanism involving autocomplementary interactions of its loop. Disruption of either DIS locus individually reduced both the yield and the thermal stability of the in vitro dimers, whereas disruption of both eliminated dimerization altogether. Similarly, the thermal stability of virion-derived dimers was impaired by deletion of both DIS elements, and point mutations in either element produced defects in viral replication that correlated with their effects on in vitro RNA dimerization. These findings support the view that in some retroviruses, dimer initiation and stability involve two or more closely linked DIS loci which together align the nascent dimer strands in parallel and in register. PMID- 11884539 TI - Regions and activities of simian virus 40 T antigen that cooperate with an activated ras oncogene in transforming primary rat embryo fibroblasts. AB - Prolonged expression of a ras oncogene in primary cells accelerates the natural process of senescence. This ras-induced permanent growth arrest is bypassed in cells expressing the simian virus 40 large T antigen. Previously we showed that two regions of T antigen, a region consisting of the N-terminal 147 amino acids and a region consisting of amino acids 251 to 708 (T251-708), independently overcome ras-induced senescence. Coexpression of either T-antigen fragment and Ras results in the appearance of dense foci of transformed cells. Using a series of mutants that produce shorter T-antigen fragments, we show that the C-terminal limit of the N-terminal T-antigen fragment that cooperates with Ras lies between amino acids 83 and 121. The N-terminal limit of the C-terminal T-antigen fragment lies between amino acids 252 and 271. In addition, we present evidence that cooperation between the N-terminal fragment and Ras depends upon an intact T antigen J domain and the ability of the T antigen to bind and inactivate the growth-suppressive effect of the tumor suppressor Rb. Introduction of specific amino acid substitutions surrounding residue 400 into T251-708 prevented the fragment from cooperating with Ras. T251-708 proteins with these same substitutions inhibited the transcriptional transactivating potential of p53 as effectively as did the wild-type protein. Thus, at least one activity contained within T251-708, other than inactivating p53 as a transcriptional transactivator, is likely to be required to bypass Ras-induced senescence. PMID- 11884540 TI - Novel immediate-early protein IE19 of human cytomegalovirus activates the origin recognition complex I promoter in a cooperative manner with IE72. AB - The major immediate-early (MIE) gene of human cytomegalovirus (HCMV) expresses IE86, IE72, IE55, and IE18 mRNA by differential splicing. Reverse transcription PCR with IE72-specific primers generated an 0.65-kb cDNA from HCMV-infected fibroblast RNA, which does not correspond to any known MIE cDNA. Nucleotide sequencing revealed that the 0.65-kb cDNA is from exons 1, 2, and 3 and part of exon 4, indicating that it is derived from a novel alternatively spliced mRNA of the MIE gene. The cDNA encodes a 172-amino-acid polypeptide, termed IE19, which corresponds to an IE72 variant with an internal deletion from Val(86) to Pro(404) and appears as a band at 38 kDa on a sodium dodecyl sulfate-polyacrylamide gel. IE19 mRNA was expressed at a low level in the immediate-early, early, and late period of viral infection. IE19 was localized in nuclei, and a transient expression assay revealed that IE19 enhances IE72-dependent activation of the HsOrc1 promoter, which is identified here as an IE72 target promoter. Another MIE protein, IE86, activated the same promoter but only weakly compared to IE72, and coexpression of IE19 did not alter the IE86-mediated transcriptional activation. In addition, IE19 did not enhance the IE72-dependent activation of the HCMV UL54 promoter. These results suggest that IE19 is a transcriptional coactivator that works with IE72. PMID- 11884541 TI - Open reading frame 50 protein of Kaposi's sarcoma-associated herpesvirus directly activates the viral PAN and K12 genes by binding to related response elements. AB - Open reading frame (ORF) 50 protein is capable of activating the entire lytic cycle of Kaposi's sarcoma-associated herpesvirus (KSHV), but its mechanism of action is not well characterized. Here we demonstrate that ORF 50 protein activates two KSHV lytic cycle genes, PAN (polyadenylated nuclear RNA) and K12, by binding to closely related response elements located approximately 60 to 100 nucleotides (nt) upstream of the start of transcription of the two genes. The 25 nt sequence 5' AAATGGGTGGCTAACCTGTCCAAAA from the PAN promoter (PANp) confers a response to ORF 50 protein in both epithelial cells and B cells in the absence of other KSHV proteins. The responsive region of DNA can be transferred to a heterologous minimal promoter. Extensive point mutagenesis showed that a span of at least 20 nt is essential for a response to ORF 50 protein. However, a minimum of six positions within this region were ambiguous. The related 26-nt responsive element in the K12 promoter (K12p), 5' GGAAATGGGTGGCTAACCCCTACATA, shares 20 nt (underlined) with the comparable region of PANp. The divergence is primarily at the 3' end. The DNA binding domain of ORF 50 protein, encompassing amino acids 1 to 490, fused to a heterologous activation domain from herpes simplex virus VP16 [ORF 50(1-490)+VP] can mediate activation of reporter constructs bearing these response elements. Most importantly, ORF 50(1-490)+VP can induce PAN RNA and K12 transcripts in transfected cells. ORF 50(1-490)+VP expressed in human cells binds specifically to duplex oligonucleotides containing the responsive regions from PANp and K12p. These DNA-protein complexes were supershifted by antibody to VP16. ORF 50(1-490) without a VP16 tag also bound to the response element. There was a strong correlation between DNA binding by ORF 50 and transcriptional activation. Mutations within PANp and K12p that impaired transactivation by ORF 50 or ORF 50(1-490)+VP also abolished DNA binding. Only one of eight related complexes formed on PANp and K12p oligonucleotides was due to ORF 50(1-490)+VP. The other complexes were due to cellular proteins. Two KSHV lytic-cycle promoters are activated by a similar mechanism that involves direct recognition of a homologous response element by the DNA binding domain of ORF 50 protein in the context of related cellular proteins. PMID- 11884543 TI - Mapping the rubella virus subgenomic promoter. AB - Rubella virus (RUB), the sole member of the Rubivirus genus in the Togaviridae family of positive-strand RNA viruses, synthesizes a single subgenomic (SG) RNA containing sequences from the 3' end of the genomic RNA including the open reading frame (ORF) that encodes the virion proteins. The synthesis of SG RNA is initiated internally on a negative-strand, genome-length template at a site known as the SG promoter (SGP). Mapping the RUB SGP was initiated by using an infectious cDNA vector, dsRobo402/GFP, in which the region containing the SGP was duplicated (K. V. Pugachev, W.-P. Tzeng, and T. K. Frey, J. Virol. 74:10811 10815, 2000). In dsRobo402/GFP, the 5'-proximal nonstructural protein ORF (NS ORF) is followed by the first SGP (SGP-1), the green fluorescent protein (GFP) gene, the second SGP (SGP-2), and the structural protein ORF. The duplicated SGP, SGP-2, contained nucleotides (nt) -175 to +76 relative to the SG start site, including the 3' 127 nt of the NS-ORF and 47 nt between the NS-ORF and the SG start site. 5' Deletions of SGP-2 to nt -40 (9 nt beyond the 3' end of the NS ORF) resulted in a wild-type (wt) phenotype in terms of virus replication and RNA synthesis. Deletions beyond this point impaired viability; however, the analysis was complicated by homologous recombination between SGP-1 and SGP-2 that resulted in deletion of the GFP gene and resurrection of viable virus with one SGP. Since the NS-ORF region was not necessary for SGP activity, subsequent mapping was done by using both replicon vectors, RUBrep/GFP and RUBrep/CAT, in which the SP-ORF is replaced with the reporter GFP and chloramphenical acetyltransferase genes, respectively, and the wt infectious clone, Robo402. In the replicon vectors, 5' deletions to nt -26 resulted in the synthesis of SG RNA. In the infectious clone, deletions through nt -28 gave rise to viable virus. A series of short internal deletions confirmed that the region between nt -28 and the SG start site was essential for viability and showed that the repeated UCA triplet at the 5' end of SG RNA was also required. Thus, the minimal SGP maps from nt -26 through the SG start site and appears to extend to at least nt +6, although a larger region is required for the generation of virus with a wt phenotype. Interestingly, while the positioning of the RUB SGP immediately adjacent the SG start site is thus similar to that of members of the genus Alphavirus, the other genus in the Togaviridae family, it does not include a region of nucleotide sequence homology with the alphavirus SGP that is located between nt -48 and nt -23 with respect to the SG start site in the RUB genome. PMID- 11884542 TI - Mechanism of human immunodeficiency virus-induced complement expression in astrocytes and neurons. AB - The cerebral complement system is hypothesized to contribute to neurodegeneration in the pathogenesis of AIDS-associated neurological disorders. Our former results have shown that the human immunodeficiency virus (HIV) strongly induces the synthesis of complement factor C3 in astrocytes. This upregulation explains in vivo data showing elevated complement levels in the cerebrospinal fluid of patients with AIDS-associated neurological symptoms. Since inhibition of complement synthesis and activation in the brain may represent a putative therapeutic goal to prevent virus-induced damage, we analyzed in detail the mechanisms of HIV-induced modulation of C3 expression. HIV-1 increased the C3 levels in astrocyte culture supernatants from 30 to up to 400 ng/ml; signal transduction studies revealed that adenylate cyclase activation with upregulation of cyclic AMP is the central signaling pathway to mediate that increase. Furthermore, activity of protein kinase C is necessary for HIV induction of C3, since inhibition of protein kinase C by prolonged exposure to the phorbol ester tetradecanoyl phorbol acetate partly abolished the HIV effect. The cytokines tumor necrosis factor alpha and gamma interferon were not involved in mediating the HIV-induced C3 upregulation, since neutralizing antibodies had no effect. Besides whole HIV virions, the purified viral proteins Nef and gp41 are biologically active in upregulating C3, whereas Tat, gp120, and gp160 were not able to modulate C3 synthesis. Further experiments revealed that neurons were also able to respond on incubation with HIV with increased C3 synthesis, although the precise pattern was slightly different from that in astrocytes. This strengthens the hypothesis that HIV-induced complement synthesis represents an important mechanism for the pathogenesis of AIDS in the brain. PMID- 11884544 TI - Lack of both Fas ligand and perforin protects from flavivirus-mediated encephalitis in mice. AB - The mechanism by which encephalitic flaviviruses enter the brain to inflict a life-threatening encephalomyelitis in a small percentage of infected individuals is obscure. We investigated this issue in a mouse model for flavivirus encephalitis in which the virus was administered to 6-week-old animals by the intravenous route, analogous to the portal of entry in natural infections, using a virus dose in the range experienced following the bite of an infectious mosquito. In this model, infection with 0.1 to 10(5) PFU of virus gave mortality in approximately 50% of animals despite low or undetectable virus growth in extraneural tissues. We show that the cytolytic effector functions play a crucial role in invasion of the encephalitic flavivirus into the brain. Mice deficient in either the granule exocytosis- or Fas-mediated pathway of cytotoxicity showed delayed and reduced mortality. Mice deficient in both cytotoxic effector functions were resistant to a low-dose peripheral infection with the neurotropic virus. PMID- 11884545 TI - In adenovirus type 12 tumorigenic cells, major histocompatibility complex class I transcription shutoff is overcome by induction of NF-kappaB and relief of COUP TFII repression. AB - The surface levels of major histocompatibility complex class I antigens are diminished on tumorigenic adenovirus type 12 (Ad12)-transformed cells, enabling them to escape from immunosurveillant cytotoxic T lymphocytes (CTLs). This is due to the down-regulation of the class I transcriptional enhancer, in which there is strong binding of the repressor COUP-TFII and lack of binding of the activator NF kappaB. Even though NF-kappaB (p65/p50) translocates to the nuclei of Ad12 transformed cells, it fails to bind to DNA efficiently due to the hypophosphorylation of the p50 subunit. In this study, tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta) were shown to promote degradation of the NF-kappaB cytoplasmic inhibitor IkappaBalpha and permit the nuclear translocation of a phosphorylated form of NF-kappaB that is capable of binding DNA. Interestingly, when Ad12-transformed cells were treated with TNF alpha or IL-1beta, class I gene transcription substantially increased when transcriptional repression by COUP-TFII was blocked. This indicates that in cytokine-treated Ad12-transformed cells, COUP-TFII is able to repress activation of class I transcription by newly nucleus-localized NF-kappaB. Our results suggest that Ad12 likely employs a "fail-safe" mechanism to ensure that the transcription of class I genes remains tightly repressed under various physiological conditions, thus providing tumorigenic Ad12-transformed cells with a means of escaping CTL recognition and lysis. PMID- 11884546 TI - Coding sequences upstream of the human immunodeficiency virus type 1 reverse transcriptase domain in Gag-Pol are not essential for incorporation of the Pr160(gag-pol) into virus particles. AB - Incorporation of the human immunodeficiency virus type 1 (HIV-1) Gag-Pol into virions is thought to be mediated by the N-terminal Gag domain via interaction with the Gag precursor. However, one recent study has demonstrated that the murine leukemia virus Pol can be incorporated into virions independently of Gag Pol expression, implying a possible interaction between the Pol and Gag precursor. To test whether the HIV-1 Pol can be incorporated into virions on removal of the N-terminal Gag domain and to define sequences required for the incorporation of Gag-Pol into virions in more detail, a series of HIV Gag-Pol expression plasmids with various extensive deletions in the region upstream of the reverse transcriptase (RT) domain was constructed, and viral incorporation of the Gag-Pol deletion mutants was examined by cotransfecting 293T cells with a plasmid expressing Pr55(gag). Analysis indicated that deletion of the N-terminal two-thirds of the gag coding region did not significantly affect the incorporation of Gag-Pol into virions. In contrast, Gag-Pol proteins with deletions covering the capsid (CA) major homology regions and the adjacent C terminal CA regions were impaired with respect to assembly into virions. However, Gag-Pol with sequences deleted upstream of the protease, or of the RT domain but retaining 15 N-terminal gag codons, could still be rescued into virions at a level about 20% of the wild-type level. When assayed in a nonmyristylated Gag-Pol context, all of the Gag-Pol deletion mutants were incorporated into virions at a level comparable to their myristylated counterparts, suggesting that the incorporation of the Gag-Pol deletion mutants into virions is independent of the N-terminal myristylation signal. PMID- 11884547 TI - Experimental inoculation of conventional pigs with porcine reproductive and respiratory syndrome virus and porcine circovirus 2. AB - Postweaning multisystemic wasting syndrome (PMWS) is a disease of nursery and fattening pigs characterized by growth retardation, paleness of the skin, dyspnea, and increased mortality rates. Porcine circovirus 2 (PCV2) has been demonstrated to be the cause of PMWS. However, other factors are needed for full development of the syndrome, and porcine reproductive and respiratory syndrome virus (PRRSV) infection has been suggested to be one of them. Twenty-four conventional 5-week-old pigs were distributed in four groups: control (n = 5), PRRSV inoculated (n = 5), PCV2 inoculated (n = 7), and PRRSV and PCV2 inoculated (n = 7). The two groups inoculated with PRRSV showed growth retardation. Pigs inoculated with both PRRSV and PCV2 had increased rectal temperature. One of these pigs developed wasting, had severe respiratory distress, and died. The most important microscopic lesion in pigs inoculated with PCV2 was lymphocyte depletion with histiocytic infiltration of the lymphoid organs, more severe and in a wider range of tissues in doubly inoculated pigs. Interstitial pneumonia was observed in the three inoculated groups. PCV2 nucleic acid was found by in situ hybridization in larger amounts and in a wider range of lymphoid tissues in PRRSV and PCV2-inoculated than in PCV2-inoculated pigs. TaqMan PCR was performed to quantify the PCV2 loads in serum during the experiment. PCV2 loads were higher in doubly inoculated pigs than in pigs inoculated with PCV2 alone. These findings indicate that severe disease can be reproduced in conventional 5-week-old pigs by inoculation of PRRSV and PCV2. Moreover, these results support the hypothesis that PRRSV infection enhances PCV2 replication. PMID- 11884548 TI - Ty5 gag mutations increase retrotransposition and suggest a role for hydrogen bonding in the function of the nucleocapsid zinc finger. AB - The Ty5 retrotransposon of Saccharomyces paradoxus transposes in Saccharomyces cerevisiae at frequencies 1,000-fold lower than do the native Ty1 elements. The low transposition activity of Ty5 could be due to differences in cellular environments between these yeast species or to naturally occurring mutations in Ty5. By screening of a Ty5 mutant library, two single mutants (D252N and Y68C) were each found to increase transposition approximately sixfold. When combined, transposition increased 36-fold, implying that the two mutations act independently. Neither mutation affected Ty5 protein synthesis, processing, cDNA recombination, or target site choice. However, cDNA levels in both single mutants and the double mutant were significantly higher than in the wild type. The D252N mutation resides in the zinc finger of nucleocapsid and increases the potential for hydrogen bonding with nucleic acids. We generated other mutations that increase the hydrogen bonding potential (i.e., D252R and D252K) and found that they similarly increased transposition. This suggests that hydrogen bonding within the zinc finger motif is important for cDNA production and builds upon previous studies implicating basic amino acids flanking the zinc finger as important for zinc finger function. Although NCp zinc fingers differ from the zinc finger motifs of cellular enzymes, the requirement for efficient hydrogen bonding is likely universal. PMID- 11884549 TI - The M184V mutation reduces the selective excision of zidovudine 5'-monophosphate (AZTMP) by the reverse transcriptase of human immunodeficiency virus type 1. AB - The M184V mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) causes resistance to lamivudine, but it also increases the sensitivity of the virus to zidovudine (3'-azido-3'-deoxythymidine; AZT). This sensitization to AZT is seen both in the presence and the absence of the mutations that confer resistance to AZT. AZT resistance is due to enhanced excision of AZT 5'-monophosphate (AZTMP) from the end of the primer by the RT of the resistant virus. Published data suggest that the excision reaction involves pyrophosphorolysis but that the likely in vivo pyrophosphate donor is not pyrophosphate but ATP. The mutations that lead to AZT resistance enhance ATP binding and, in so doing, enhance pyrophosphorolysis. The excision reaction is specific for AZT because HIV-1 RT, which can form a closed complex with a dideoxy terminated primer and an incoming deoxynucleoside triphosphate (dNTP), does not form the closed complex with an AZTMP-terminated primer and an incoming dNTP. This means that an AZTMP-terminated primer has better access to the site where it can be excised. The M184V mutation alters the polymerase active site in a fashion that specifically interferes with ATP-mediated excision of AZTMP from the end of the primer strand. The M184V mutation does not affect the incorporation of AZT 5' triphosphate (AZTTP), either in the presence or the absence of mutations that enhance AZTMP excision. However, in the presence of ATP, the M184V mutation does decrease the ability of HIV-1 RT to carry out AZTMP excision. Based on these results, and on the results of other excision experiments, we present a model to explain how the M184V mutation affects AZTMP excision. PMID- 11884550 TI - Interaction between parvovirus NS2 protein and nuclear export factor Crm1 is important for viral egress from the nucleus of murine cells. AB - A mutation that disrupts the interaction between the NS2 protein of minute virus of mice and the nuclear export factor Crm1 results in a block to egress of mutant generated full virions from the nucleus of infected murine cells. These mutants produce wild-type levels of monomer and dimer replicative DNA forms but are impaired in their ability to generate progeny single-stranded DNA in restrictive murine cells in the first round of infection. The NS2-Crm1 interaction mutant can be distinguished phenotypically from an NS2-null mutant and reveals a role for the Crm1-mediated export pathway at a late step in viral infection. PMID- 11884551 TI - The fusion peptide of Semliki Forest virus associates with sterol-rich membrane domains. AB - Semliki Forest virus (SFV) is an enveloped alphavirus whose membrane fusion is triggered by low pH and promoted by cholesterol and sphingolipid in the target membrane. Fusion is mediated by E1, a viral membrane protein containing the putative fusion peptide. Virus mutant studies indicate that SFV's cholesterol dependence is controlled by regions of E1 outside of the fusion peptide. Both E1 and E1*, a soluble ectodomain form of E1, interact with membranes in a reaction dependent on low pH, cholesterol, and sphingolipid and form highly stable homotrimers. Here we have used detergent extraction and gradient floatation experiments to demonstrate that E1* associated selectively with detergent resistant membrane domains (DRMs or rafts). In contrast, reconstituted full length E1 protein or influenza virus fusion peptide was not associated with DRMs. Methyl beta-cyclodextrin quantitatively extracted both cholesterol and E1* from membranes in the absence of detergent, suggesting a strong association of E1* with sterol. Monoclonal antibody studies demonstrated that raft association was mediated by the proposed E1 fusion peptide. Thus, although other regions of E1 are implicated in the control of virus cholesterol dependence, once the SFV fusion peptide inserts in the target membrane it has a high affinity for membrane domains enriched in cholesterol and sphingolipid. PMID- 11884552 TI - Distribution of spontaneous mutants and inferences about the replication mode of the RNA bacteriophage phi6. AB - When a parent virus replicates inside its host, it must first use its own genome as the template for replication. However, once progeny genomes are produced, the progeny can in turn act as templates. Depending on whether the progeny genomes become templates, the distribution of mutants produced by an infection varies greatly. While information on the distribution is important for many population genetic models, it is also useful for inferring the replication mode of a virus. We have analyzed the distribution of mutants emerging from single bursts in the RNA bacteriophage phi6 and find that the distribution closely matches a Poisson distribution. The match suggests that replication in this bacteriophage is effectively by a stamping machine model in which the parental genome is the main template used for replication. However, because the distribution deviates slightly from a Poisson distribution, the stamping machine is not perfect and some progeny genomes must replicate. By fitting our data to a replication model in which the progeny genomes become replicative at a given rate or probability per round of replication, we estimated the rate to be very low and on the on the order of 10(-4). We discuss whether different replication modes may confer an adaptive advantage to viruses. PMID- 11884553 TI - Herpes simplex virus tegument protein US11 interacts with conventional kinesin heavy chain. AB - Little is known about the mechanisms of transport of neurotropic herpesviruses, such as herpes simplex virus (HSV), varicella-zoster virus, and pseudorabies virus, within neurons. For these viruses, which replicate in the nucleus, anterograde transport from the cell body of dorsal root ganglion (DRG) neurons to the axon terminus occurs over long distances. In the case of HSV, unenveloped nucleocapsids in human DRG neurons cocultured with autologous skin were observed by immunoelectron microscopy to colocalize with conventional ubiquitous kinesin, a microtubule-dependent motor protein, in the cell body and axon during anterograde axonal transport. Subsequently, four candidate kinesin-binding structural HSV proteins were identified (VP5, VP16, VP22, and US11) using oligohistidine-tagged human ubiquitous kinesin heavy chain (uKHC) as bait. Of these viral proteins, a direct interaction between uKHC and US11 was identified. In vitro studies identified residues 867 to 894 as the US11-binding site in uKHC located within the proposed heptad repeat cargo-binding domain of uKHC. In addition, the uKHC-binding site in US11 maps to the C-terminal RNA-binding domain. US11 is consistently cotransported with kinetics similar to those of the capsid protein VP5 into the axons of dissociated rat neurons, unlike the other tegument proteins VP16 and VP22. These observations suggest a major role for the uKHC-US11 interaction in anterograde transport of unenveloped HSV nucleocapsids in axons. PMID- 11884554 TI - Nuclear interactions are necessary for translational enhancement by spleen necrosis virus RU5. AB - The 5' long terminal repeat of spleen necrosis virus (SNV) facilitates Rev/Rev responsive element (RRE)-independent expression of intron-containing human immunodeficiency virus type 1 (HIV-1) gag. The SNV RU5 region, which corresponds to the 165-nucleotide 5' RNA terminus, functions in a position- and orientation dependent manner to enhance polysome association of intron-containing HIV-1 gag RNA and also nonviral luc RNA. Evidence is mounting that association with nuclear factors during intron removal licenses mRNAs for nuclear export, efficient translation, and nonsense-mediated decay. This project addressed the relationship between the nuclear export pathway of SNV RU5-reporter RNA and translational enhancement. Results of RNA transfection experiments suggest that cytoplasmic proteins are insufficient for SNV RU5 translational enhancement of gag or luc RNA. Reporter gene assays, leptomycin B (LMB) sensitivity experiments, and RNase protection assays indicate that RU5 gag RNA accesses a nuclear export pathway that is distinct from the LMB-inhibited leucine-rich nuclear export sequence dependent CRM1 pathway, which is used by the HIV-1 RRE. As a unique tool with which to investigate the relationship between different RNA trafficking routes and translational enhancement, SNV RU5 and Rev/RRE were combined on a single gag RNA. We observed a less-than-synergistic effect on cytoplasmic mRNA utilization. Instead, Rev/RRE diverts RU5 gag RNA to the CRM1-dependent, LMB-inhibited pathway and abrogates translational enhancement by SNV RU5. Our study is the first to show that a nuclear factor(s) directs SNV RU5-containing RNAs to a distinct export pathway that is not inhibited by LMB and programs the intron-containing transcript for translational enhancement. PMID- 11884555 TI - The RNA binding domain of the hantaan virus N protein maps to a central, conserved region. AB - The nucleocapsid (N) protein of hantaviruses encapsidates both viral genomic and antigenomic RNAs, although only the genomic viral RNA (vRNA) is packaged into virions. To define the domain within the Hantaan virus (HTNV) N protein that mediates these interactions, 14 N- and C-terminal deletion constructs were cloned into a bacterial expression vector, expressed, and purified to homogeneity. Each protein was examined for its ability to bind the HTNV S segment vRNA with filter binding and gel electrophoretic mobility shift assays. These studies mapped a minimal region within the HTNV N protein (amino acids 175 to 217) that bound vRNA. Sequence alignments made from several hantavirus N protein sequences showed that the region identified has a 58% identity and an 86% similarity among these amino acid sequences. Two peptides corresponding to amino acids 175 to 196 (N1) and 197 to 218 (N2) were synthesized. The RNA binding of each peptide was measured by filter binding and competition analysis. Three oligoribonucleotides were used to measure binding affinity and assess specificity. The N2 peptide contained the major RNA binding determinants, while the N1 peptide, when mixed with N2, contributed to the specificity of vRNA recognition. PMID- 11884556 TI - Induction of mucosal protection against primary, heterologous simian immunodeficiency virus by a DNA vaccine. AB - An effective vaccine against human immunodeficiency virus (HIV) should protect against mucosal transmission of genetically divergent isolates. As a safe alternative to live attenuated vaccines, the immunogenicity and protective efficacy of a DNA vaccine containing simian immunodeficiency virus (SIV) strain 17E-Fr (SIV/17E-Fr) gag-pol-env was analyzed in rhesus macaques. Significant levels of cytotoxic T lymphocytes (CTL), but low to undetectable serum antibody responses, were observed following multiple immunizations. SIV-specific mucosal antibodies and CTL were also detected in rectal washes and gut-associated lymphoid tissues, respectively. Vaccinated and naive control monkeys were challenged intrarectally with SIV strain DeltaB670 (SIV/DeltaB670), a primary isolate whose env is 15% dissimilar to that of the vaccine strain. Four of seven vaccinees were protected from infection as determined by the inability to identify viral RNA or DNA sequences in the peripheral blood and the absence of anamnestic antibody responses postchallenge. This is the first report of mucosal protection against a primary pathogenic, heterologous isolate of SIV by using a commercially viable vaccine approach. These results support further development of a DNA vaccine for protection against HIV. PMID- 11884557 TI - Derivation and characterization of a dengue type 1 host range-restricted mutant virus that is attenuated and highly immunogenic in monkeys. AB - We recently described the derivation of a dengue serotype 2 virus (DEN2mutF) that exhibited a host range-restricted phenotype; it was severely impaired for replication in cultured mosquito cells (C6/36 cells). DEN2mutF virus had selected mutations in genomic sequences predicted to form a 3' stem-loop structure (3'-SL) that is conserved among all flavivirus species. The 3'-SL constitutes the downstream terminal similar95 nucleotides of the 3' noncoding region in flavivirus RNA. Here we report the introduction of these same mutational changes into the analogous region of an infectious DNA derived from the genome of a human virulent dengue serotype 1 virus (DEN1), strain Western Pacific (DEN1WP). The resulting DEN1 mutant (DEN1mutF) exhibited a host range-restricted phenotype similar to that of DEN2mutF virus. DEN1mutF virus was attenuated in a monkey model for dengue infection in which viremia is taken as a correlate of human virulence. In spite of the markedly reduced levels of viremia that it induced in monkeys compared to DEN1WP, DEN1mutF was highly immunogenic. In addition, DEN1mutF-immunized monkeys retained high levels of neutralizing antibodies in serum and were protected from challenge with high doses of the DEN1WP parent for as long as 17 months after the single immunizing dose. Phenotypic revertants of DEN1mutF and DEN2mutF were each detected after a total of 24 days in C6/36 cell cultures. Complete nucleotide sequence analysis of DEN1mutF RNA and that of a revertant virus, DEN1mutFRev, revealed that (i) the DEN1mutF genome contained no additional mutations upstream from the 3'-SL compared to the DEN1WP parent genome and (ii) the DEN1mutFRev genome contained de novo mutations, consistent with our previous hypothesis that the defect in DEN2mutF replication in C6/36 cells was at the level of RNA replication. A strategy for the development of a tetravalent dengue vaccine is discussed. PMID- 11884558 TI - Recombinant vaccinia virus-induced T-cell immunity: quantitation of the response to the virus vector and the foreign epitope. AB - Recombinant vaccinia viruses (rVV) have been extensively used as vaccines, but there is little information about the total magnitude of the VV-specific T-cell response and how this compares to the immune response to the foreign gene(s) expressed by the rVV. To address this issue, we quantitated the T-cell responses to both the viral vector and the insert following the infection of mice with VV expressing a cytotoxic T lymphocyte (CTL) epitope (NP118-126) from lymphocytic choriomeningitis virus (LCMV). The LCMV epitope-specific response was quantitated by intracellular cytokine staining after stimulation with the specific peptide. To analyze the total VV-specific response, we developed a simple intracellular cytokine staining assay using VV-infected major histocompatibility complex class I and II matched cells as stimulators. Using this approach, we made the following determinations. (i) VV-NP118 induced potent and long-lasting CD8 and CD4 T-cell responses to the vector; at the peak of the response (approximately 1 week), there were approximately 10(7) VV-specific CD8 T cells (25% of the CD8 T cells) and approximately 10(6) VV-specific CD4 T cells (approximately 5% of the CD4 T cells) in the spleen. These numbers decreased to approximately 5 x 10(5) CD8 T cells (approximately 5% frequency) and approximately 10(5) CD4 T cells (approximately 0.5% frequency), respectively, by day 30 and were then stably maintained at these levels for >300 days. The size of this VV-specific T-cell response was comparable to that of the T-cell response induced following an acute LCMV infection. (ii) VV-specific CD8 and CD4 T cells were capable of producing gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and interleukin-2; all cells were able to make IFN-gamma, a subset produced both IFN gamma and TNF-alpha, and another subset produced all three cytokines. (iii) The CD8 T-cell response to the foreign gene (LCMV NP118-126 epitope) was coordinately regulated with the response to the vector during all three phases (expansion, contraction, and memory) of the T-cell response. The total number of CD8 T cells responding to NP118-126 were approximately 20- to 30-fold lower than the number responding to the VV vector (approximately 1% at the peak and 0.2% in memory). This study provides a better understanding of T-cell immunity induced by VV-based vaccines, and in addition, the technique described in the study can be readily extended to other viral vectors to determine the ratio of the T-cell response to the insert versus the vector. This information will be useful in optimizing prime boost regimens for vaccination. PMID- 11884559 TI - Mutation of the dominant endocytosis motif in human immunodeficiency virus type 1 gp41 can complement matrix mutations without increasing Env incorporation. AB - The human immunodeficiency virus type 1 transmembrane glycoprotein (TM) is efficiently endocytosed in a clathrin-dependent manner. Internalization is mediated by a tyrosine-containing motif within the cytoplasmic domain, and replacement of the cytoplasmic tyrosine by cysteine or phenylalanine increased expression of mutant glycoprotein on the surface of transfected cells by as much as 2.5-fold. Because interactions between the cytoplasmic domain of Env and the matrix protein (MA) have been suggested to mediate incorporation of Env in virus particles, we examined whether perturbation of endocytosis would alter incorporation. Proviruses were constructed to contain the wild-type or mutant Env in conjunction with point mutations in MA that had previously been shown to block Env incorporation. These constructs were used to evaluate the effect of glycoprotein endocytosis on incorporation into virus particles and to test the necessity for a specific interaction between Env and MA to mediate incorporation. Viruses produced from transfected 293T cells were used to infect various cell lines, including MAGI, H9, and CEMx174. Viruses encoding both a disrupted endocytosis motif signal and mutations within MA were significantly more infectious in MAGI cells than their counterparts encoding a mutant MA and wild type Env. This complementation of infectivity for the MA incorporation mutant viruses was not due to increased glycoprotein incorporation into particles but instead reflected an enhanced fusogenicity of the mutated Env proteins. Our findings further support the concept that a specific interaction between the long cytoplasmic domain of TM and MA is required for efficient incorporation of Env into assembling virions. Alteration of the endocytosis signal of Env, and the resulting increase in cell surface glycoprotein, has no effect on incorporation despite demonstrable effects on fusion, virus entry, and infectivity. PMID- 11884560 TI - Stable replication of papillomavirus genomes in Saccharomyces cerevisiae. AB - Papillomaviruses normally replicate in stratified squamous epithelial tissues of their mammalian hosts, in which the viral genome is found as a nuclear plasmid. Two viral proteins, E1, a helicase, and E2, a transcriptional activator and plasmid maintenance factor, are known to contribute to the episomal replication of the viral genome. Recently, our laboratory discovered that papillomaviruses can also replicate in an E1-independent manner in mammalian cells (K. Kim and P. F. Lambert, Virology, in press; K. Kim and P. F. Lambert, submitted for publication). In this study, we describe experiments investigating the capacity of the human papillomavirus type 16 (HPV16) genome to replicate in yeast (Saccharomyces cerevisiae). The full-length HPV16 genome, when linked in cis to a selectable yeast marker gene, either TRP1 or URA3, could replicate stably as an episome in yeast. The replication of papillomavirus genomes in yeast is not limited to HPV16. Bovine papillomavirus type 1 and HPV6b, -11, -16, -18, and -31 were all capable of replicating in short-term assays over a period of 20 cell doublings. The long-term persistence of viral episomes did not require any one viral gene, as mutant genomes defective in single genes also replicated episomally. These results indicate that the viral episome can replicate in the absence of the E1 DNA helicase. Similarly, E2 was also not required for replication in yeast, and E2 mutant viral genomes were stably maintained in the absence of selection, indicating the existence of an E2-independent mechanism for plasmid maintenance. The episomal replication of papillomavirus genomes in yeast provides a genetically manipulatable system in which to investigate cellular factors required for episomal replication and may provide a novel means for generating infectious papillomavirus. PMID- 11884561 TI - Replication of bovine papillomavirus type 1 (BPV-1) DNA in Saccharomyces cerevisiae following infection with BPV-1 virions. AB - Saccharomyces cerevisiae protoplasts exposed to bovine papillomavirus type 1 (BPV 1) virions demonstrated uptake of virions on electron microscopy. S. cerevisiae cells looked larger after exposure to BPV-1 virions, and cell wall regeneration was delayed. Southern blot hybridization of Hirt DNA from cells exposed to BPV-1 virions demonstrated BPV-1 DNA, which could be detected over 80 days of culture and at least 13 rounds of division. Two-dimensional gel analysis of Hirt DNA showed replicative intermediates, confirming that the BPV-1 genome was replicating within S. cerevisiae. Nicked circle, linear, and supercoiled BPV-1 DNA species were observed in Hirt DNA preparations from S. cerevisiae cells infected for over 50 days, and restriction digestion showed fragments hybridizing to BPV-1 in accord with the predicted restriction map for circular BPV-1 episomes. These data suggest that BPV-1 can infect S. cerevisiae and that BPV-1 episomes can replicate in the infected S. cerevisiae cells. PMID- 11884563 TI - Overexpression of the rabies virus glycoprotein results in enhancement of apoptosis and antiviral immune response. AB - A recombinant rabies virus (RV) carrying two identical glycoprotein (G) genes (SPBNGA-GA) was constructed and used to determine the effect of RV G overexpression on cell viability and immunity. Immunoprecipitation analysis and flow cytometry showed that tissue culture cells infected with SPBNGA-GA produced, on average, twice as much RV G as cells infected with RV carrying only a single RV G gene (SPBNGA). The overexpression of RV G in SPBNGA-GA-infected NA cells was paralleled by a significant increase in caspase 3 activity followed by a marked decrease in mitochondrial respiration, neither of which was observed in SPBNGA infected cells. Furthermore, fluorescence staining and confocal microscopy revealed an increased extent of apoptosis and markedly reduced neurofilament and F actin in SPBNGA-GA-infected primary neuron cultures compared with neuronal cells infected with SPBNGA, supporting the concept that RV G or motifs of the RV G gene trigger the apoptosis cascade. Mice immunized with SPBNGA-GA showed substantially higher antibody titers against the RV G and against the nucleoprotein than SPBNGA-immunized mice, suggesting that the speed or extent of apoptosis directly determines the magnitude of the antibody response. PMID- 11884564 TI - Evolutionary history of Cucumber mosaic virus deduced by phylogenetic analyses. AB - Cucumber mosaic virus (CMV) is an RNA plant virus with a tripartite genome and an extremely broad host range. Previous evolutionary analyses with the coat protein (CP) and 5' nontranslated region (NTR) of RNA 3 suggested subdivision of the virus into three groups, subgroups IA, IB, and II. In this study 15 strains of CMV whose nucleotide sequences have been determined were used for a complete phylogenetic analysis of the virus. The trees estimated for open reading frames (ORFs) located on the different RNAs were not congruent and did not completely support the subgrouping indicated by the CP ORF, indicating that different RNAs had independent evolutionary histories. This is consistent with a reassortment mechanism playing an important role in the evolution of the virus. The evolutionary trees of the 1a and 3a ORFs were more compact and displayed more branching than did those of the 2a and CP ORFs. This may reflect more rigid host interactive constraints exerted on the 1a and 3a ORFs. In addition, analysis of the 3' NTR that is conserved among all RNAs indicated that evolutionary constraints on this region are specific to the RNA component rather than the virus isolate. This indicates that functions other than replication are encoded in the 3' NTR. Reassortment may have led to the genetic diversity found among CMV strains and contributed to its enormous evolutionary success. PMID- 11884562 TI - Coxsackievirus B3 replication is reduced by inhibition of the extracellular signal-regulated kinase (ERK) signaling pathway. AB - Coxsackievirus B3 (CVB3) is the most common human pathogen for viral myocarditis. We have previously shown that the signaling protein p21(ras) GTPase-activating protein (RasGAP) is cleaved and that mitogen-activated protein kinases (MAPKs) ERK1/2 are activated in the late phase of CVB3 infection. However, the role of intracellular signaling pathways in CVB3-mediated myocarditis and the relative advantages of such pathways to host or virus remain largely unclear. In this study we extended our prior studies by examining the interaction between CVB3 replication and intracellular signaling pathways in HeLa cells. We observed that CVB3 infection induced a biphasic activation of ERK1/2, early transient activation versus late sustained activation, which were regulated by different mechanisms. Infection by UV-irradiated, inactivated virus capable of receptor binding and endocytosis triggered early ERK1/2 activation, but was insufficient to trigger late ERK1/2 activation. By using a general caspase inhibitor (zVAD.fmk) we further demonstrated that late ERK1/2 activation was not a result of CVB3-mediated caspase cleavage. Treatment of cells with U0126, a selective inhibitor of MAPK kinase (MEK), significantly inhibited CVB3 progeny release and decreased virus protein production. Furthermore, inhibition of ERK1/2 activation circumvented CVB3-induced apoptosis and viral protease-mediated RasGAP cleavage. Taken together, these data suggest that ERK1/2 activation is important for CVB3 replication and contributes to virus-mediated changes in host cells. Our findings demonstrate coxsackievirus takeover of a particular host signaling mechanism and uncover a prospective approach to stymie virus spread and preserve myocardial integrity. PMID- 11884565 TI - Intra- and intercellular trafficking of the foamy virus auxiliary bet protein. AB - The Bet protein of foamy viruses (FVs) is an auxiliary protein encoded by the 3' end of the viral genome. Although its function during the viral replication cycle is still unknown, Bet seems to play a key role in the establishment and/or maintenance of viral persistence, representing the predominant viral protein detected during chronic infection. To clarify the function of this viral protein, the subcellular distribution of Bet from the prototypic human foamy virus (HFV) was examined. We report here that this protein is distributed in both the cytoplasm and the nucleus of HFV-infected or Bet-transfected cells. The nuclear targeting results from the presence of a bipartite nuclear localization signal at the C-terminal region, sufficient to direct heterologous reporter proteins to the nucleus. Since HFV Bet spreads between cells, we show here that the secreted protein targets the nuclei of recipient cells. HFV Bet follows an unconventional route to exit the cell since its secretion is not affected by brefeldin A, a drug which disrupts the trafficking between the endoplasmic reticulum and the Golgi complex. Finally, these inter- and intracellular movements were also observed for the equine foamy virus Bet protein, strongly suggesting that these remarkable features are conserved among FVs. PMID- 11884566 TI - Altered patterns of cellular gene expression in dermal microvascular endothelial cells infected with Kaposi's sarcoma-associated herpesvirus. AB - Kaposi's sarcoma (KS)-associated herpesvirus (KSHV; also called human herpesvirus 8) is believed to be the etiologic agent of Kaposi's sarcoma, multicentric Castleman's disease, and AIDS-associated primary effusion lymphoma. KSHV infection of human dermal microvascular endothelial cells (DMVEC) in culture results in the conversion of cobblestone-shaped cells to spindle-shaped cells, a characteristic morphological feature of cells in KS lesions. All spindle-shaped cells in KSHV-infected DMVEC cultures express the latency-associated nuclear protein LANA1, and a subfraction of these cells undergo spontaneous lytic cycle induction that can be enhanced by tetradecanoyl phorbol acetate (TPA) treatment. To study the cellular response to infection by KSHV, we used two different gene array screening systems to examine the expression profile of either 2,350 or 9,180 human genes in infected compared to uninfected DMVEC cultures in both the presence and absence of TPA. In both cases, between 1.4 and 2.5% of the genes tested were found to be significantly upregulated or downregulated. Further analysis by both standard and real-time reverse transcription-PCR procedures directly confirmed these results for 14 of the most highly upregulated and 13 of the most highly downregulated genes out of a total of 37 that were selected for testing. These included strong upregulation of interferon-responsive genes such as interferon response factor 7 (IRF7) and myxovirus resistance protein R1, plus upregulation of exodus 2 beta-chemokine, RDC1 alpha-chemokine receptor, and transforming growth factor beta3, together with strong downregulation of cell adhesion factors alpha(4)-integrin and fibronectin plus downregulation of bone morphogenesis protein 4, matrix metalloproteinase 2, endothelial plasminogen activator inhibitor 1, connective tissue growth factor, and interleukin-8. Significant dysregulation of several other cytokine-related genes or receptors, as well as endothelial cell and macrophage markers, and various other genes associated with angiogenesis or transformation was also detected. Western immunoblot and immunohistochemical analyses confirmed that the cellular IRF7 protein levels were strongly upregulated during the early lytic cycle both in KSHV-infected DMVEC and in the body cavity-based lymphoma BCBL1 PEL cell line. PMID- 11884568 TI - E1 protein of bovine papillomavirus type 1 interferes with E2 protein-mediated tethering of the viral DNA to mitotic chromosomes. AB - Eukaryotic viruses can maintain latency in dividing cells as extrachromosomal plasmids. It is therefore of vital importance for viruses to ensure nuclear retention and proper segregation of their viral DNA. The bovine papillomavirus (BPV) E2 enhancer protein plays a key role in these processes by tethering the viral DNA to the host cell chromosomes. Viral genomes that harbor phosphorylation mutations in the E2 gene are transformation defective, and for these mutant genomes, neither the viral DNA nor the E2 protein is detected on mitotic chromosomes, while other key functions of E2 in transcription and replication were wild type. Moreover, secondary mutations in both the E2 and E1 proteins lead to suppression of the phosphorylation mutant phenotype and resulted in reattachment of the viral DNA and the E2 protein onto mitotic chromosomes, suggesting that E1 also plays a role in viral genome partitioning. The E1 protein was cytologically always excluded from mitotic chromatin, either as a suppressor allele or as the wild type. In the absence of other viral proteins, an E2 protein containing alanine substitutions for phosphorylation substrates in the hinge region (E2-A4) was detected as wild-type on mitotic chromosomes. However, when wild-type E1 protein levels were increased in cells expressing either the A4 mutant E2 proteins or wild-type E2, the E2-A4 protein was much more sensitive to chromosomal dislocation than was the wild-type protein. In contrast, suppressor alleles of E1 were not capable of such abrogation of E2 binding (A4 or wild-type) to chromosomes. These results suggest that wild-type E1 can be a negative regulator of the chromosomal attachment of E2. PMID- 11884567 TI - Patterns of gene expression and a transactivation function exhibited by the vGCR (ORF74) chemokine receptor protein of Kaposi's sarcoma-associated herpesvirus. AB - The ORF74 or vGCR gene encoded by Kaposi's sarcoma-associated herpesvirus (KSHV; also called human herpesvirus 8) has properties of a ligand-independent membrane receptor signaling protein with angiogenic properties that is predicted to play a key role in the biology of the virus. We have examined the expression of vGCR mRNA and protein in primary effusion lymphoma (PEL) cell lines, PEL and multicentric Castleman's disease (MCD) tumors, Kaposi's sarcoma lesions and infected endothelial cell cultures. The vGCR gene proved to be expressed in PEL cell lines as a large spliced bicistronic mRNA of 3.2 kb that also encompasses the upstream vOX2 (K14) gene. This mRNA species was induced strongly by phorbol ester (TPA) and sodium butyrate treatment in the BCBL-1 cell line, but only weakly in the HBL6 cell line, and was classified as a relatively late and low abundance delayed early class lytic cycle gene product. A complex bipartite upstream lytic cycle promoter for this mRNA was nestled within the intron of the 5'-overlapping but oppositely oriented latent-state transcription unit for LANA1/vCYC-D/vFLIP and responded strongly to both TPA induction and cotransfection with the KSHV RNA transactivator protein (RTA or ORF50) in transient reporter gene assays. A vGCR protein product of 45 kDa that readily dimerized was detected by Western blotting and in vitro translation and was localized in a cytoplasmic and membrane pattern in DNA-transfected Vero and 293T cells or adenovirus vGCR-transduced dermal microvascular endothelial cells (DMVEC) as detected by indirect immunofluorescence assay (IFA) and immunohistochemistry with a specific rabbit anti-vGCR antibody. Similarly, a subfraction of KSHV-positive cultured PEL cells and of KSHV (JSC-1) persistently infected DMVEC cells displayed cytoplasmic vGCR protein expression, but only after TPA or spontaneous lytic cycle induction, respectively. The vGCR protein was also detectable by immunohistochemical staining in a small fraction (0.5 to 3%) of the cells in PEL and MCD tumor and nodular Kaposi's sarcoma lesion specimens that were apparently undergoing lytic cycle expression. These properties are difficult to reconcile with the vGCR protein's playing a direct role in spindle cell proliferation, transformation, or latency, but could be compatible with proposed contributions to angiogenesis via downstream paracrine effects. The ability of vGCR to transactivate expression of both several KSHV promoter-driven luciferase (LUC) reporter genes and an NFkappaB motif containing the chloramphenicol acetyltransferase (CAT) reporter gene may also suggest an unexpected regulatory role in viral gene expression. PMID- 11884569 TI - Adenovirus binding to the coxsackievirus and adenovirus receptor or integrins is not required to elicit brain inflammation but is necessary to transduce specific neural cell types. AB - Intracranial administration of adenovirus vectors elicits rapid, capsid-mediated dose-dependent brain inflammation. The mechanisms through which adenovirus capsids trigger inflammation in the brain remain unknown. We determined whether adenovirus interaction with the primary and secondary cell surface receptors for infection (CAR and alphav integrins) was necessary to trigger acute adenovirus mediated brain inflammation, and, furthermore, whether capsid mutations that abrogated CAR and integrin binding altered vector tropism in the brain. Vectors ablated for CAR binding, but retaining integrin binding function, transduced equivalent areas of brain compared to vectors with wild-type capsids; however, vector tropsim was dramatically altered. Vectors with wild-type capsids predominantly transduced oligodendrocytes, whereas mutation of the fiber protein to ablate CAR binding resulted in a loss of oligodendrocyte transduction and a consequent redirection of transduction to neurons and other types of glial cells. Combined mutations of fiber and penton base that ablate both CAR and integrin binding almost abolished brain transduction. Thus, doubly-ablated capsids engineered to express new ligands should allow complete vector retargeting in the central nervous system. Although transduction by the doubly-ablated vectors was reduced by greater than 95%, inflammation was not reduced compared to wild-type vectors, demonstrating that brain inflammation occurs independently of adenovirus binding and infection of cells via CAR and integrin receptors. PMID- 11884570 TI - Rotavirus NSP5: mapping phosphorylation sites and kinase activation and viroplasm localization domains. AB - Rotavirus NSP5 is a nonstructural protein that localizes in cytoplasmic viroplasms of infected cells. NSP5 interacts with NSP2 and undergoes a complex posttranslational hyperphosphorylation, generating species with reduced polyacrylamide gel electrophoresis mobility. This process has been suggested to be due in part to autophosphorylation. We developed an in vitro phosphorylation assay using as a substrate an in vitro-translated NSP5 deletion mutant that was phosphorylated by extracts from MA104 cells transfected with NSP5 mutants but not by extracts from mock-transfected cells. The phosphorylated products obtained showed shifts in mobility similar to what occurs in vivo. From these and other experiments we concluded that NSP5 activates a cellular kinase(s) for its own phosphorylation. Three NSP5 regions were found to be essential for kinase(s) activation. Glutathione S-transferase-NSP5 mutants were produced in Escherichia coli and used to determine phosphoacceptor sites. These were mapped to four serines (Ser(153), Ser(155), Ser(163), and Ser(165)) within an acidic region with homology to casein kinase II (CKII) phosphorylation sites. CKII was able to phosphorylate NSP5 in vitro. NSP5 and its mutants fused to enhanced green fluorescent protein were used in transfection experiments followed by virus infection and allowed the determination of the domains essential for viroplasm localization in the context of virus infection. PMID- 11884571 TI - Evidence of a role for nonmuscle myosin II in herpes simplex virus type 1 egress. AB - After cell entry, herpes simplex virus (HSV) particles are transported through the host cell cytoplasm to nuclear pores. Following replication, newly synthesized virus particles are transported back to the cell periphery via a complex pathway including a cytoplasmic phase involving some form of unenveloped particle. These various transport processes are likely to make use of one or more components of the cellular cytoskeletal systems and associated motor proteins. Here we report that the HSV type 1 (HSV-1) major tegument protein, VP22, interacts with the actin-associated motor protein nonmuscle myosin IIA (NMIIA). HSV-1 infection resulted in reorganization of NMIIA, inducing retraction of NMIIA from the cell periphery and condensation into a spoke-like distribution around the nucleus along with a second effect of accumulation in a perinuclear cluster. VP22 did not appear to colocalize with the reorganized cagelike distribution of NMIIA. However, VP22 has been previously reported to localize in a perinuclear vesicular pattern, and significant overlap was observed between this pattern and the perinuclear clusters of NMIIA. Inhibition of the ATPase activity of NMIIA with the myosin-specific inhibitor butanedione monoxime impaired the formation of the perinuclear vesicular VP22 accumulations and also the release of virus into the extracellular medium while having much less effect on the yield of cell associated virus. Virus infection frequently results in the induction of highly extended processes emanating from the infected cell, and we observed that VP22 containing particles line up along NMIIA-containing filaments which run through these protrusions. PMID- 11884572 TI - Structural analysis of the hepatitis C virus RNA polymerase in complex with ribonucleotides. AB - We report here the results of a systematic high-resolution X-ray crystallographic analysis of complexes of the hepatitis C virus (HCV) RNA polymerase with ribonucleoside triphosphates (rNTPs) and divalent metal ions. An unexpected observation revealed by this study is the existence of a specific rGTP binding site in a shallow pocket at the molecular surface of the enzyme, 30 A away from the catalytic site. This previously unidentified rGTP pocket, which lies at the interface between fingers and thumb, may be an allosteric regulatory site and could play a role in allowing alternative interactions between the two domains during a possible conformational change of the enzyme required for efficient initiation. The electron density map at 1.7-A resolution clearly shows the mode of binding of the guanosine moiety to the enzyme. In the catalytic site, density corresponding to the triphosphates of nucleotides bound to the catalytic metals was apparent in each complex with nucleotides. Moreover, a network of triphosphate densities was detected; these densities superpose to the corresponding moieties of the nucleotides observed in the initiation complex reported for the polymerase of bacteriophage phi6, strengthening the proposal that the two enzymes initiate replication de novo by similar mechanisms. No equivalent of the protein stacking platform observed for the priming nucleotide in the phi6 enzyme is present in HCV polymerase, however, again suggesting that a change in conformation of the thumb domain takes place upon template binding to allow for efficient de novo initiation of RNA synthesis. PMID- 11884573 TI - Activation of nuclear factor of activated T cells by human T-lymphotropic virus type 1 accessory protein p12(I). AB - Human T-lymphotropic virus type 1 (HTLV-1) is the agent of an aggressive malignancy of CD4(+) T lymphocytes, called adult T-cell lymphoma/leukemia, and is associated with numerous immune-mediated diseases. To establish infection, HTLV-1 must activate targeted T cells during early stages of infection. We recently demonstrated that the HTLV-1 accessory protein p12(I) is critical for persistent infection in vivo and for viral infectivity in quiescent primary lymphocytes, suggesting a role for p12(I) in lymphocyte activation. To test whether p12(I) modulates signaling pathways required for T-lymphocyte activation, we examined AP 1-, NF-kappaB-, and nuclear factor of activated T cells (NFAT)-driven reporter gene activity in p12(I)-expressing Jurkat T cells compared to vector-transfected control cells. HTLV-1 p12(I) specifically induced NFAT-mediated transcription approximately 20-fold in synergy with the Ras/mitogen-activated protein kinase pathway, but did not influence AP-1- or NF-kappaB-dependent gene expression. Inhibition of calcium-dependent signals by cyclosporin A, BAPTA-AM [glycine, N,N' 1,2-ethanediylbis(oxy-2,1-phenylene)-bis-N-2-(acetyloxy)methoxy-2-oxoethyl] [bis(acetyloxy)methyl ester], and a dominant negative mutant of NFAT2 abolished the p12(I)-mediated activation of NFAT-dependent transcription. In contrast, inhibition of phospholipase C-gamma and LAT (linker for activation of T cells) did not affect p12(I)-induced NFAT activity. Importantly, p12(I) functionally substituted for thapsigargin, which selectively depletes intracellular calcium stores. Our data are the first to demonstrate a role for HTLV-1 p12(I) in calcium dependent activation of NFAT-mediated transcription in lymphoid cells. We propose a novel mechanism by which HTLV-1, a virus associated with lymphoproliferative disease, dysregulates common T-cell activation pathways critical for the virus to establish persistent infection. PMID- 11884574 TI - Mapping of genes involved in murine herpes simplex virus keratitis: identification of genes and their modifiers. AB - Herpes simplex keratitis (HSK) is an inflammatory response to viral infection and self antigens in the cornea and is a major cause of blindness. Using two strains of mice which are susceptible (129/SVEV) and resistant (C57BL/6) to herpes simplex virus (HSV) strain KOS, (129/SVEV x C57BL/6)F(2) mice were generated and examined for their disease susceptibility in terms of clinical symptoms, ocular disease, and antibody production following corneal scarification with HSV (KOS). A genome-wide screen was carried out using microsatellite markers to determine the genetic loci involved in this response. Loci on chromosomes 4, 5, 12, 13, and 14 were shown to be involved in general susceptibility to clinical disease, whereas loci on chromosomes 10 and 17 were shown to be unique to ocular disease. PMID- 11884576 TI - Altering expression levels of human immunodeficiency virus type 1 gp120-gp41 affects efficiency but not kinetics of cell-cell fusion. AB - Human immunodeficiency virus (HIV) entry into a host cell requires the fusion of virus and cellular membranes that is driven by interaction of the viral envelope glycoproteins gp120 and gp41 (gp120/gp41) with CD4 and a coreceptor, typically either CXCR4 or CCR5. The stoichiometry of gp120/gp41:CD4:CCR5 necessary to initiate membrane fusion is not known. To allow an examination of early events in gp120/gp41-driven membrane fusion, we developed a novel real-time cell-cell fusion assay. Using this assay to study fusion kinetics, we found that altering the cell surface density of gp120/gp41 affected the maximal extent of fusion without dramatically altering fusion kinetics. Collectively, these observations are consistent with the view that gp120/gp41-driven membrane fusion requires the formation of a threshold number of fusion-active intercellular gp120/gp41:CD4:CCR5 complexes. Furthermore, the probability of reaching this threshold is governed, in part, by the surface density of gp120/gp41. PMID- 11884575 TI - Solid-phase proteoliposomes containing human immunodeficiency virus envelope glycoproteins. AB - The human immunodeficiency virus type 1 (HIV-1) exterior envelope glycoprotein gp120 mediates receptor binding and is the major target for neutralizing antibodies. A broadly neutralizing antibody response is likely to be a critical component of the immune response against HIV-1. Although antibodies against monomeric gp120 are readily elicited in immunized individuals, these antibodies are inefficient in neutralizing primary HIV-1 isolates. As a chronic pathogen, HIV-1 has evolved to avoid an optimal host response by a number of immune escape mechanisms. Monomeric gp120 that has dissociated from the functional trimer presents irrelevant epitopes that are not accessible on functional trimeric envelope glycoproteins. The resulting low level of antigenic cross-reactivity between monomeric gp120 and the functional spike may contribute to the inability of monomeric gp120 to elicit broadly neutralizing antibodies. Attempts to generate native, trimeric envelope glycoproteins as immunogens have been frustrated by both the lability of the gp120-gp41 interaction and the weak association between gp120 subunits. Here, we present solid-phase HIV-1 gp160DeltaCT (cytoplasmic tail-deleted) proteoliposomes (PLs) containing native, trimeric envelope glycoproteins in a physiologic membrane setting. We present data that indicate that the gp160DeltaCT glycoproteins on PLs are trimers and are recognized by several relevant conformational ligands in a manner similar to that for gp160DeltaCT oligomers expressed on the cell surface. The PLs represent a significant advance over present envelope glycoprotein formulations as candidate immunogens for HIV vaccine design and development. PMID- 11884577 TI - Capsid protein C of tick-borne encephalitis virus tolerates large internal deletions and is a favorable target for attenuation of virulence. AB - Deletions ranging in size from 4 to 21 amino acid residues were introduced into the capsid protein of the flavivirus tick-borne encephalitis (TBE) virus. These deletions incrementally affected a hydrophobic domain which is present at the center of all flavivirus capsid protein sequences and part of which may form an amphipathic alpha-helix. In the context of the full-length TBE genome, the deletions did not measurably affect protein expression and up to a deletion length of 16 amino acid residues, corresponding to almost 17% of mature protein C, viable virus was recovered. This virus was strongly attenuated but highly immunogenic in adult mice, revealing capsid protein C as a new and attractive target for the directed attenuation of flaviviruses. Apparently, the larger deletions interfered with the correct assembly of infectious virus particles, and this disturbance of virion assembly is likely to be the molecular basis of attenuation. However, all of the mutants carrying large deletions produced substantial amounts of subviral particles, which as judged from density gradient analyses were identical to recombinant subviral particles as obtained by the expression of the surface proteins prM and E alone. The structural and functional flexibility of protein C revealed in this study and its predicted largely alpha helical conformation are reminiscent of capsid proteins of other enveloped viruses, such as alphaviruses (N-terminal domain of the capsid protein), retroviruses, and hepadnaviruses and suggest that all of these may belong to a common structural class, which is fundamentally distinct from the classical beta barrel structures of many icosahedral viral capsids. The possibility of attenuating flaviviruses by disturbing virus assembly and favoring the production of noninfectious but highly immunogenic subviral particles opens up a promising new avenue for the development of live flavivirus vaccines. PMID- 11884579 TI - Heterologous movement protein strongly modifies the infection phenotype of cucumber mosaic virus. AB - A hybrid virus (CMVcymMP) constructed by replacing the movement protein (MP) of cucumber mosaic cucumovirus (CMV) with that of cymbidium ringspot tombusvirus (CymRSV) was viable and could efficiently spread both cell to cell and long distance in host plants. The hybrid virus was able to move cell to cell in the absence of functional CP, whereas CP-deficient CMV was restricted to single inoculated cells. In several Chenopodium and Nicotiana species, the symptom phenotype of the hybrid virus infection was clearly determined by the foreign MP gene. In Nicotiana debneyi and Nicotiana tabacum cv. Xanthi, the hybrid virus could move systemically, contrary to CymRSV. PMID- 11884578 TI - Apical budding of a recombinant influenza A virus expressing a hemagglutinin protein with a basolateral localization signal. AB - Influenza virions bud preferentially from the apical plasma membrane of infected epithelial cells, by enveloping viral nucleocapsids located in the cytosol with its viral integral membrane proteins, i.e., hemagglutinin (HA), neuraminidase (NA), and M2 proteins, located at the plasma membrane. Because individually expressed HA, NA, and M2 proteins are targeted to the apical surface of the cell, guided by apical sorting signals in their transmembrane or cytoplasmic domains, it has been proposed that the polarized budding of influenza virions depends on the interaction of nucleocapsids and matrix proteins with the cytoplasmic domains of HA, NA, and/or M2 proteins. Since HA is the major protein component of the viral envelope, its polarized surface delivery may be a major force that drives polarized viral budding. We investigated this hypothesis by infecting MDCK cells with a transfectant influenza virus carrying a mutant form of HA (C560Y) with a basolateral sorting signal in its cytoplasmic domain. C560Y HA was expressed nonpolarly on the surface of infected MDCK cells. Interestingly, viral budding remained apical in C560Y virus-infected cells, and so did the location of NP and M1 proteins at late times of infection. These results are consistent with a model in which apical viral budding is a shared function of various viral components rather than a role of the major viral envelope glycoprotein HA. PMID- 11884580 TI - Redirecting retroviral tropism by insertion of short, nondisruptive peptide ligands into envelope. AB - A potentially powerful approach for in vivo gene delivery is to target retrovirus to specific cells through interactions between cell surface receptors and appropriately modified viral envelope proteins. Previously, relatively large (>100 residues) protein ligands to cell surface receptors have been inserted at or near the N terminus of retroviral envelope proteins. Although viral tropism could be altered, the chimeric envelope proteins lacked full activity, and coexpression of wild-type envelope was required for production of transducing virus. Here we analyze more than 40 derivatives of ecotropic Moloney murine leukemia virus (MLV) envelope, containing insertions of short RGD-containing peptides, which are ligands for integrin receptors. In many cases pseudotyped viruses containing only the chimeric envelope protein could transduce human cells. The precise location, size, and flanking sequences of the ligand affected transduction specificity and efficiency. We conclude that retroviral tropism can be rationally reengineered by insertion of short peptide ligands and without the need to coexpress wild-type envelope. PMID- 11884581 TI - Selective targeting and inducible destruction of human cancer cells by retroviruses with envelope proteins bearing short peptide ligands. AB - In the accompanying study, we show how retroviral tropism can be redirected by insertion of short peptide ligands at multiple locations in envelope. Here we use this approach to selectively target and destroy human cancer cells. Many cancer cells overexpress specific cell surface receptors. We have generated Moloney murine leukemia virus (MLV) envelope derivatives bearing short peptide ligands for gastrin-releasing protein (GRP) and human epidermal growth factor receptors. Pseudotyped viruses containing these chimeric envelope derivatives selectively transduce human cancer cell lines that overexpress the cognate receptor. A retrovirus targeting the GRP receptor can deliver the thymidine kinase gene to human melanoma and breast cancer cells, which are killed by the subsequent addition of ganciclovir. Collectively, our results demonstrate that short peptide ligands inserted at appropriate locations in MLV envelope can selectively target retroviruses to human cancer cells and deliver a therapeutically relevant gene. PMID- 11884583 TI - Varicella-zoster virus open reading frame 2 encodes a membrane phosphoprotein that is dispensable for viral replication and for establishment of latency. AB - Varicella-zoster virus (VZV) encodes six genes that do not have homologs in herpes simplex virus. One of these genes, VZV open reading frame 2 (ORF2), was expressed as a 31-kDa phosphoprotein in the membranes of infected cells. Unlike equine and bovine herpesvirus type 1 ORF2 homologs that are associated with virions, VZV virions contained no detectable ORF2 protein. The ORF2 deletion mutant established a latent infection in cotton rats at a frequency and with a number of VZV genomes similar to that of the parental virus. ORF63 transcripts, a hallmark of latent infection, were present in ganglia latently infected with both the ORF2 deletion mutant and parental VZV. Thus, ORF2 is the first VZV gene shown to be dispensable for establishment of latent infection in an animal model. PMID- 11884582 TI - Inhibition of hepatitis B virus replication by interferon requires proteasome activity. AB - Hepatitis B virus (HBV) replication is inhibited in a noncytopathic manner by alpha/beta interferon (IFN-alpha/beta) and IFN-gamma. We demonstrate here that inhibitors of cellular proteasome activity can block this antiviral effect. These results suggest that a critical component of the IFN-induced antiviral response may be the proteasome-dependent degradation of viral or cellular proteins that are required for HBV replication. PMID- 11884584 TI - Inhibition of in vitro leukocyte proliferation by morbilliviruses. AB - Immune suppression associated with morbillivirus infections may influence the mortality rate by allowing secondary bacterial infections that are lethal to the host to flourish. Using an in vitro proliferation assay, we have shown that all members of the genus Morbillivirus inhibit the proliferation of a human B lymphoblast cell line (BJAB). Proliferation of freshly isolated, stimulated bovine and caprine peripheral blood lymphocytes is also inhibited by UV inactivated rinderpest (RPV) and peste-des-petits ruminants viruses. As for measles virus, coexpression of both the fusion and the hemagglutinin proteins of RPV is necessary and sufficient to induce immune suppression in vitro. PMID- 11884585 TI - The PHD type zinc finger is an integral part of the CBP acetyltransferase domain. AB - Histone acetyltransferases (HATs) such as CBP and p300 are regarded as key regulators of RNA polymerase II-mediated transcription, but the critical structural features of their HAT modules remain ill defined. The HAT domains of CBP and p300 are characterized by the presence of a highly conserved putative plant homeodomain (PHD) (C4HC3) type zinc finger, which is part of the functionally uncharacterized cysteine-histidine-rich region 2 (CH2). Here we show that this region conforms to the PHD type zinc finger consensus and that it is essential for in vitro acetylation of core histones and the basal transcription factor TFIIE34 as well as for CBP autoacetylation. PHD finger mutations also reduced the transcriptional activity of the full-length CBP protein when tested on transfected reporter genes. Importantly, similar results were obtained on integrated reporters, which reflect a more natural chromatinized state. Taken together, our results indicate that the PHD finger forms an integral part of the enzymatic core of the HAT domain of CBP. PMID- 11884586 TI - Ctr9, Rtf1, and Leo1 are components of the Paf1/RNA polymerase II complex. AB - The Saccharomyces cerevisiae Paf1-RNA polymerase II (Pol II) complex is biochemically and functionally distinct from the Srb-mediator form of Pol II holoenzyme and is required for full expression of a subset of genes. In this work we have used tandem affinity purification tags to isolate the Paf1 complex and mass spectrometry to identify additional components. We have established that Ctr9, Rtf1, and Leo1 are factors that associate with Paf1, Cdc73, and Pol II, but not with the Srb-mediator. Deletion of either PAF1 or CTR9 leads to similar severe pleiotropic phenotypes, which are unaltered when the two mutations are combined. In contrast, we found that deletion of LEO1 or RTF1 leads to few obvious phenotypes, although mutation of RTF1 suppresses mutations in TATA binding protein, alters transcriptional start sites, and affects elongation. Remarkably, deletion of LEO1 or RTF1 suppresses many paf1Delta phenotypes. In particular, an rtf1Delta paf1Delta double mutant grew faster, was less temperature sensitive, and was more resistant to caffeine and hydroxyurea than a paf1Delta single mutant. In addition, expression of the G(1) cyclin CLN1, reduced nearly threefold in paf1Delta, is restored to wild-type levels in the rtf1Delta paf1Delta double mutant. We suggest that lack of Paf1 results in a defective complex and a block in transcription, which is relieved by removal of Leo1 or Rtf1. PMID- 11884588 TI - Targeted disruption of the testicular SPAG5/deepest protein does not affect spermatogenesis or fertility. AB - In an effort to define the molecular basis for morphogenesis of major sperm tail structures, including outer dense fibers, we recently cloned the Spag5 gene by virtue of its strong and specific leucine-zipper-mediated interaction with Odf1, the 27-kDa major outer dense fiber protein. Spag5 is expressed during meiosis and in round spermatids and is similar, if not identical, to Deepest, a putative spindle pole protein. Here we report the disruption of the Spag5 gene by homologous recombination. Spag5-null mice lack Spag5 mRNA and protein. However, male mice are viable and fertile. Analysis of the process of spermatogenesis and sperm produced in Spag5-null mice did not reveal a major phenotype as a consequence of the knockout event. This result suggests that if Spag5 plays a role in spermatogenesis it is likely compensated for by unknown proteins. PMID- 11884587 TI - Stat1-dependent, p53-independent expression of p21(waf1) modulates oxysterol induced apoptosis. AB - 7-Ketocholesterol (7kchol) is prominent in atherosclerotic lesions where apoptosis occurs. Using mouse fibroblasts lacking p53, p21(waf1), or Stat1, we found that optimal 7kchol-induced apoptosis requires p21(waf1) and Stat1 but not p53. Findings were analogous in a human cell system. Apoptosis was restored in Stat1-null human cells when wild-type Stat1 was restored. Phosphorylation of Stat1 on Ser(727) but not Tyr(701) was essential for optimum apoptosis. A neutralizing antibody against beta interferon (IFN-beta) blunted Ser(727) phosphorylation and apoptosis after 7kchol treatment; cells deficient in an IFN beta receptor subunit exhibited blunted apoptosis. IFN-beta alone did not induce apoptosis; thus, 7kchol-induced release of IFN-beta was necessary but not sufficient for optimal apoptosis. In Stat1-null cells, expression of p21(waf1) was much less than in wild-type cells; introducing transient expression of p21(waf1) restored apoptosis. Stat1 and p21(waf1) were essential for downstream apoptotic events, including cytochrome c release from mitochondria and activation of caspases 9 and 3. Our data reveal key elements of the cellular pathway through which an important oxysterol induces apoptosis. Identification of the essential signaling events that may pertain in vivo could suggest targets for therapeutic intervention. PMID- 11884589 TI - Regulation of insulin-like growth factor type I (IGF-I) receptor kinase activity by protein tyrosine phosphatase 1B (PTP-1B) and enhanced IGF-I-mediated suppression of apoptosis and motility in PTP-1B-deficient fibroblasts. AB - The insulin-like growth factor type I (IGF-I) receptor (IGF-IR), activated by its ligands IGF-I and IGF-II, can initiate several signal transduction pathways that mediate suppression of apoptosis, proliferation, differentiation, and transformation. Here we investigated the regulation of IGF-IR activation and function by protein tyrosine phosphatase 1B (PTP-1B). Coexpression of PTP-1B with a beta-chain construct of the IGF-IR (betaWT) inhibited IGF-IR kinase activity in fission yeast Schizosaccharomyces pombe, in COS cells, and in IGF-IR-deficient fibroblasts. In both spontaneously immortalized and simian virus 40 T antigen transformed embryonic fibroblast cell lines derived from PTP-1B knockout mice, IGF-I induced higher levels of IGF-IR autophosphorylation and kinase activity than were induced in PTP-1B-expressing control cells. PTP-1B-deficient cells exhibited enhanced IGF-I-mediated protection from apoptosis in response to serum withdrawal or etoposide killing, as well as enhanced plating efficiency and IGF-I mediated motility. Reexpression of PTP-1B in spontaneously immortalized fibroblasts resulted in decreased IGF-IR and AKT activation, as well as decreased protection from apoptosis and decreased motility. These findings demonstrate that PTP-1B can regulate IGF-IR kinase activity and function and that loss of PTP-1B can enhance IGF-I-mediated cell survival, growth, and motility in transformed cells. PMID- 11884590 TI - Proteomics analysis reveals stable multiprotein complexes in both fission and budding yeasts containing Myb-related Cdc5p/Cef1p, novel pre-mRNA splicing factors, and snRNAs. AB - Schizosaccharomyces pombe Cdc5p and its Saccharomyces cerevisiae ortholog, Cef1p, are essential Myb-related proteins implicated in pre-mRNA splicing and contained within large multiprotein complexes. Here we describe the tandem affinity purification (TAP) of Cdc5p- and Cef1p-associated complexes. Using transmission electron microscopy, we show that the purified Cdc5p complex is a discrete structure. The components of the S. pombe Cdc5p/S. cerevisiae Cef1p complexes (termed Cwfs or Cwcs, respectively) were identified using direct analysis of large protein complex (DALPC) mass spectrometry (A. J. Link et al., Nat. Biotechnol. 17:676-682, 1999). At least 26 proteins were detected in the Cdc5p/Cef1p complexes. Comparison of the polypeptides identified by S. pombe Cdc5p purification with those identified by S. cerevisiae Cef1p purification indicates that these two yeast complexes are nearly identical in composition. The majority of S. pombe Cwf proteins and S. cerevisiae Cwc proteins are known pre mRNA splicing factors including core Sm and U2 and U5 snRNP components. In addition, the complex contains the U2, U5, and U6 snRNAs. Previously uncharacterized proteins were also identified, and we provide evidence that several of these novel factors are involved in pre-mRNA splicing. Our data represent the first comprehensive analysis of CDC5-associated proteins in yeasts, describe a discrete highly conserved complex containing novel pre-mRNA splicing factors, and demonstrate the power of DALPC for identification of components in multiprotein complexes. PMID- 11884591 TI - Control of cell cycle exit and entry by protein kinase B-regulated forkhead transcription factors. AB - AFX-like Forkhead transcription factors, which are controlled by phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) signaling, are involved in regulating cell cycle progression and cell death. Both cell cycle arrest and induction of apoptosis are mediated in part by transcriptional regulation of p27(kip1). Here we show that the Forkheads AFX (FOXO4) and FKHR-L1 (FOXO3a) also directly control transcription of the retinoblastoma-like p130 protein and cause upregulation of p130 protein expression. Detailed analysis of p130 regulation demonstrates that following Forkhead-induced cell cycle arrest, cells enter G(0) and become quiescent. This is shown by a change in phosphorylation of p130 to G(0)-specific forms and increased p130/E2F-4 complex formation. Most importantly, long-term Forkhead activation causes a sustained but reversible inhibition of proliferation without a marked increase in apoptosis. As for the activity of the Forkheads, we also show that protein levels of p130 are controlled by endogenous PI3K/PKB signaling upon cell cycle reentry. Surprisingly, not only nontransformed cells, but also cancer cells such as human colon carcinoma cells, are forced into quiescence by Forkhead activation. We therefore propose that Forkhead inactivation by PKB signaling in quiescent cells is a crucial step in cell cycle reentry and contributes to the processes of transformation and regeneration. PMID- 11884593 TI - Ime2, a meiosis-specific kinase in yeast, is required for destabilization of its transcriptional activator, Ime1. AB - In the budding yeast Saccharomyces cerevisiae, entry into meiosis and its successful completion depend on two positive regulators, Ime1 and Ime2. Ime1 is a transcriptional activator that is required for transcription of IME2, a serine/threonine protein kinase. We show that in vivo Ime2 associates with Ime1, that in vitro Ime2 phosphorylates Ime1, and that in living cells the stability of Ime1 depends on Ime2. Diploid cells with IME2 deleted show an increase in the level of Ime1, whereas haploid cells overexpressing IME2 show a decrease in the stability of Ime1. Furthermore, the level of Ime1 depends on the kinase activity of Ime2. Using a mutation in one of the ATPase subunits of the proteasome, RPT2, we demonstrate that Ime1, amino acids 270 to 360, is degraded by the 26S proteasome. We also show that Ime2 itself is an extremely unstable protein whose expression in vegetative cultures is toxic. We propose that a negative-feedback loop ensures that the activity of Ime1 will be restricted to a narrow window. PMID- 11884592 TI - Partial reconstitution of human DNA mismatch repair in vitro: characterization of the role of human replication protein A. AB - DNA mismatch repair (MMR) is a critical genome-stabilization system. However, the molecular mechanism of MMR in human cells remains obscure because many of the components have not yet been identified. Using a functional in vitro reconstitution system, this study identified three HeLa cell fractions essential for in vitro MMR. These fractions divide human MMR into two distinct stages: mismatch-provoked excision and repair synthesis. In vitro dissection of the MMR reaction and crucial intermediates elucidated biochemical functions of individual fractions in human MMR and identified hitherto unknown functions of human replication protein A (hRPA) in MMR. Thus, one fraction carries out nick-directed and mismatch-dependent excision; the second carries out DNA repair synthesis and DNA ligation; and the third provides hRPA, which plays multiple roles in human MMR by protecting the template DNA strand from degradation, enhancing repair excision, and facilitating repair synthesis. It is anticipated that further analysis of these fractions will identify additional MMR components and enable the complete reconstitution of the human MMR pathway with purified proteins. PMID- 11884595 TI - Targeted transposition by the V(D)J recombinase. AB - Cleavage by the V(D)J recombinase at a pair of recombination signal sequences creates two coding ends and two signal ends. The RAG proteins can integrate these signal ends, without sequence specificity, into an unrelated target DNA molecule. Here we demonstrate that such transposition events are greatly stimulated by--and specifically targeted to--hairpins and other distorted DNA structures. The mechanism of target selection by the RAG proteins thus appears to involve recognition of distorted DNA. These data also suggest a novel mechanism for the formation of alternative recombination products termed hybrid joints, in which a signal end is joined to a hairpin coding end. We suggest that hybrid joints may arise by transposition in vivo and propose a new model to account for some recurrent chromosome translocations found in human lymphomas. According to this model, transposition can join antigen receptor loci to partner sites that lack recombination signal sequence elements but bear particular structural features. The RAG proteins are capable of mediating all necessary breakage and joining events on both partner chromosomes; thus, the V(D)J recombinase may be far more culpable for oncogenic translocations than has been suspected. PMID- 11884594 TI - CRM1-dependent function of a cis-acting RNA export element. AB - Viruses often contain cis-acting RNA elements, which facilitate the posttranscriptional processing and export of their messages. These elements fall into two classes distinguished by the presence of either viral or cellular RNA binding proteins. To date, studies have indicated that the viral proteins utilize the CRM1-dependent export pathway, while the cellular factors generally function in a CRM1-independent manner. The cis-acting element found in the woodchuck hepatitis virus (WHV) (the WHV posttranscriptional regulatory element [WPRE]) has the ability to posttranscriptionally stimulate transgene expression and requires no viral proteins to function. Conventional wisdom suggests that the WPRE would function in a CRM1-independent manner. However, our studies on this element reveal that its efficient function is sensitive to the overexpression of the C terminus of CAN/Nup214 and treatment with the antimicrobial agent leptomycin B. Furthermore, the overexpression of CRM1 stimulates WPRE activity. These results suggest a direct role for CRM1 in the export function of the WPRE. This observation suggests that the WPRE is directing messages into a CRM1-dependent mRNA export pathway in somatic mammalian cells. PMID- 11884596 TI - Differential effects of chromatin and Gcn4 on the 50-fold range of expression among individual yeast Ty1 retrotransposons. AB - Approximately 30 copies of the Ty1 retrotransposon are present in the genome of Saccharomyces cerevisiae. Previous studies gave insights into the global regulation of Ty1 transcription but provided no information on the behavior of individual genomic elements. This work shows that the expression of 31 individual Ty1 elements in S288C varies over a 50-fold range. Their transcription is repressed by chromatin structures, which are antagonized by the Swi/Snf and SAGA chromatin-modifying complexes in highly expressed Ty1 elements. These elements carry five potential Gcn4 binding sites in their promoter regions that are mostly absent in weakly expressed Ty1 copies. Consistent with this observation, Gcn4 activates the transcription of highly expressed Ty1 elements only. One of the potential Gcn4 binding sites acts as an upstream activating sequence in vivo and interacts with Gcn4 in vitro. Since Gcn4 has been shown to interact with Swi/Snf and SAGA, we predict that Gcn4 activates Ty1 transcription by targeting these complexes to specific Ty1 promoters. PMID- 11884597 TI - Shared role for differentially methylated domains of imprinted genes. AB - For most imprinted genes, a difference in expression between the maternal and paternal alleles is associated with a corresponding difference in DNA methylation that is localized to a differentially methylated domain (DMD). Removal of a gene's DMD leads to a loss of imprinting. These observations suggest that DMDs have a determinative role in genomic imprinting. To examine this possibility, we introduced sequences from the DMDs of the imprinted Igf2r, H19, and Snrpn genes into a nonimprinted derivative of the normally imprinted RSVIgmyc transgene, created by excising its own DMD. Hybrid transgenes with sequences from the Igf2r DMD2 were consistently imprinted, with the maternal allele being more methylated than the paternal allele. Only the repeated sequences within DMD2 were required for imprinting these transgenes. Hybrid transgenes containing H19 and Snrpn DMD sequences and ones containing sequences from the long terminal repeat of a murine intracisternal A particle retrotransposon were not imprinted. The Igf2r hybrid transgenes are comprised entirely of mouse genomic DNA and behave as endogenous imprinted genes in inbred wild-type and mutant mouse strains. These types of hybrid transgenes can be used to elucidate the functions of DMD sequences in genomic imprinting. PMID- 11884598 TI - Convergence of multiple signaling cascades at glycogen synthase kinase 3: Edg receptor-mediated phosphorylation and inactivation by lysophosphatidic acid through a protein kinase C-dependent intracellular pathway. AB - Lysophosphatidic acid (LPA) is a natural phospholipid with multiple biological functions. We show here that LPA induces phosphorylation and inactivation of glycogen synthase kinase 3 (GSK-3), a multifunctional serine/threonine kinase. The effect of LPA can be reconstituted by expression of Edg-4 or Edg-7 in cells lacking LPA responses. Compared to insulin, LPA stimulates only modest phosphatidylinositol 3-kinase (PI3K)-dependent activation of protein kinase B (PKB/Akt) that does not correlate with the magnitude of GSK-3 phosphorylation induced by LPA. PI3K inhibitors block insulin- but not LPA-induced GSK-3 phosphorylation. In contrast, the effect of LPA, but not that of insulin or platelet-derived growth factor (PDGF), is sensitive to protein kinase C (PKC) inhibitors. Downregulation of endogenous PKC activity selectively reduces LPA mediated GSK-3 phosphorylation. Furthermore, several PKC isotypes phosphorylate GSK-3 in vitro and in vivo. To confirm a specific role for PKC in regulation of GSK-3, we further studied signaling properties of PDGF receptor beta subunit (PDGFRbeta) in HEK293 cells lacking endogenous PDGF receptors. In clones expressing a PDGFRbeta mutant wherein the residues that couple to PI3K and other signaling functions are mutated with the link to phospholipase Cgamma (PLCgamma) left intact, PDGF is fully capable of stimulating GSK-3 phosphorylation. The process is sensitive to PKC inhibitors in contrast to the response through the wild-type PDGFRbeta. Therefore, growth factors, such as PDGF, which control GSK-3 mainly through the PI3K-PKB/Akt module, possess the ability to regulate GSK-3 through an alternative, redundant PLCgamma-PKC pathway. LPA and potentially other natural ligands primarily utilize a PKC-dependent pathway to modulate GSK-3. PMID- 11884600 TI - Dnmt1 overexpression causes genomic hypermethylation, loss of imprinting, and embryonic lethality. AB - Biallelic expression of Igf2 is frequently seen in cancers because Igf2 functions as a survival factor. In many tumors the activation of Igf2 expression has been correlated with de novo methylation of the imprinted region. We have compared the intrinsic susceptibilities of the imprinted region of Igf2 and H19, other imprinted genes, bulk genomic DNA, and repetitive retroviral sequences to Dnmt1 overexpression. At low Dnmt1 methyltransferase levels repetitive retroviral elements were methylated and silenced. The nonmethylated imprinted region of Igf2 and H19 was resistant to methylation at low Dnmt1 levels but became fully methylated when Dnmt1 was overexpressed from a bacterial artificial chromosome transgene. Methylation caused the activation of the silent Igf2 allele in wild type and Dnmt1 knockout cells, leading to biallelic Igf2 expression. In contrast, the imprinted genes Igf2r, Peg3, Snrpn, and Grf1 were completely resistant to de novo methylation, even when Dnmt1 was overexpressed. Therefore, the intrinsic difference between the imprinted region of Igf2 and H19 and of other imprinted genes to postzygotic de novo methylation may be the molecular basis for the frequently observed de novo methylation and upregulation of Igf2 in neoplastic cells and tumors. Injection of Dnmt1-overexpressing embryonic stem cells in diploid or tetraploid blastocysts resulted in lethality of the embryo, which resembled embryonic lethality caused by Dnmt1 deficiency. PMID- 11884599 TI - Enumeration of the simian virus 40 early region elements necessary for human cell transformation. AB - While it is clear that cancer arises from the accumulation of genetic mutations that endow the malignant cell with the properties of uncontrolled growth and proliferation, the precise combinations of mutations that program human tumor cell growth remain unknown. The study of the transforming proteins derived from DNA tumor viruses in experimental models of transformation has provided fundamental insights into the process of cell transformation. We recently reported that coexpression of the simian virus 40 (SV40) early region (ER), the gene encoding the telomerase catalytic subunit (hTERT), and an oncogenic allele of the H-ras gene in normal human fibroblast, kidney epithelial, and mammary epithelial cells converted these cells to a tumorigenic state. Here we show that the SV40 ER contributes to tumorigenic transformation in the presence of hTERT and oncogenic H-ras by perturbing three intracellular pathways through the actions of the SV40 large T antigen (LT) and the SV40 small t antigen (ST). LT simultaneously disables the retinoblastoma (pRB) and p53 tumor suppressor pathways; however, complete transformation of human cells requires the additional perturbation of protein phosphatase 2A by ST. Expression of ST in this setting stimulates cell proliferation, permits anchorage-independent growth, and confers increased resistance to nutrient deprivation. Taken together, these observations define the elements of the SV40 ER required for the transformation of human cells and begin to delineate a set of intracellular pathways whose disruption, in aggregate, appears to be necessary to generate tumorigenic human cells. PMID- 11884602 TI - Regulation of E2F1-dependent gene transcription and apoptosis by the ETS-related transcription factor GABPgamma1. AB - The E2F family of transcription factors comprises six related members which are involved in the control of the coordinated progression through the G(1)/S-phase transition of cell cycle or in cell fate decision. Their activity is regulated by pocket proteins, including pRb, p107, and p130. Here we show that E2F1 directly interacts with the ETS-related transcription factor GABPgamma1 in vitro and in vivo. The binding domain interacting with GABPgamma1 was mapped to the C-terminal amino acids 310 to 437 of E2F1, which include its transactivation and pRb binding domain. Among the E2F family of transcription factors, the interaction with GABPgamma1 is restricted to E2F1. DNA-binding E2F1 complexes containing GABPgamma1 are characterized by enhanced E2F1-dependent transcriptional activity. Moreover, GABPgamma1 suppresses E2F1-dependent apoptosis by mechanisms other than the inhibition of the transactivation capacity of E2F1. In summary, our results provide evidence for a novel pRb-independent mechanism regulating E2F1-dependent transcription and apoptosis. PMID- 11884601 TI - Protein kinase A associates with HA95 and affects transcriptional coactivation by Epstein-Barr virus nuclear proteins. AB - HA95, a nuclear protein homologous to AKAP95, has been identified in immune precipitates of the Epstein-Barr virus (EBV) coactivating nuclear protein EBNA-LP from EBV-transformed lymphoblastoid cells (LCLs). We now find that HA95 and EBNA LP are highly associated in LCLs and in B-lymphoma cells where EBNA-LP is expressed by gene transfer. Binding was also evident in yeast two-hybrid assays. HA95 binds to the EBNA-LP repeat domain that is the principal coactivator of transcription. EBNA-LP localizes with HA95 and causes HA95 to partially relocalize with EBNA-LP in promyelocytic leukemia nuclear bodies. Protein kinase A catalytic subunit alpha (PKAcsalpha) is significantly associated with HA95 in the presence or absence of EBNA-LP. Although EBNA-LP is not a PKA substrate, HA95 or PKAcsalpha expression in B lymphoblasts specifically down-regulates the strong coactivating effects of EBNA-LP. The inhibitory effects of PKAcsalpha are reversed by coexpression of protein kinase inhibitor. PKAcsalpha also inhibits EBNA-LP coactivation with the EBNA-2 acidic domain fused to the Gal4 DNA binding domain. Furthermore, EBNA-LP- and EBNA-2-induced expression of the EBV oncogene, LMP1, is down-regulated by PKAcsalpha or HA95 expression in EBV-infected lymphoblasts. These experiments indicate that HA95 and EBNA-LP localize PKAcsalpha at nuclear sites where it can affect transcription from specific promoters. The role of HA95 as a scaffold for transcriptional regulation is discussed. PMID- 11884603 TI - Involvement of mouse Rev3 in tolerance of endogenous and exogenous DNA damage. AB - The Rev3 gene of Saccharomyces cerevisiae encodes the catalytic subunit of DNA polymerase zeta that is implicated in mutagenic translesion synthesis of damaged DNA. To investigate the function of its mouse homologue, we have generated mouse embryonic stem cells and mice carrying a targeted disruption of Rev3. Although some strain-dependent variation was observed, Rev3(-/-) embryos died around midgestation, displaying retarded growth in the absence of consistent developmental abnormalities. Rev3(-/-) cell lines could not be established, indicating a cell-autonomous requirement of Rev3 for long-term viability. Histochemical analysis of Rev3(-/-) embryos did not reveal aberrant replication or cellular proliferation but demonstrated massive apoptosis in all embryonic lineages. Although increased levels of p53 are detected in Rev3(-/-) embryos, the embryonic phenotype was not rescued by the absence of p53. A significant increase in double-stranded DNA breaks as well as chromatid and chromosome aberrations was observed in cells from Rev3(-/-) embryos. The inner cell mass of cultured Rev3(-/ ) blastocysts dies of a delayed apoptotic response after exposure to a low dose of N-acetoxy-2-acetylaminofluorene. These combined data are compatible with a model in which, in the absence of polymerase zeta, double-stranded DNA breaks accumulate at sites of unreplicated DNA damage, eliciting a p53-independent apoptotic response. Together, these data are consistent with involvement of polymerase zeta in translesion synthesis of endogenously and exogenously induced DNA lesions. PMID- 11884604 TI - Functional divergence between histone deacetylases in fission yeast by distinct cellular localization and in vivo specificity. AB - Histone deacetylases (HDACs) are important for gene regulation and the maintenance of heterochromatin in eukaryotes. Schizosaccharomyces pombe was used as a model system to investigate the functional divergence within this conserved enzyme family. S. pombe has three HDACs encoded by the hda1(+), clr3(+), and clr6(+) genes. Strains mutated in these genes have previously been shown to display strikingly different phenotypes when assayed for viability, chromosome loss, and silencing. Here, conserved differences in the substrate binding pocket identify Clr6 and Hda1 as class I HDACs, while Clr3 belongs in the class II family. Furthermore, these HDACs were shown to have strikingly different subcellular localization patterns. Hda1 was localized to the cytoplasm, while most of Clr3 resided throughout the nucleus. Finally, Clr6 was localized exclusively on the chromosomes in a spotted pattern. Interestingly, Clr3, the only HDAC present in the nucleolus, was required for ribosomal DNA (rDNA) silencing. Clr3 presumably acts directly on heterochromatin, since it colocalized with the centromere, mating-type region, and rDNA as visualized by in situ hybridization. In addition, Clr3 could be cross-linked to mat3 in chromatin immunoprecipitation experiments. Western analysis of bulk histone preparations indicated that Hda1 (class I) had a generally low level of activity in vivo and Clr6 (class I) had a high level of activity and broad in vivo substrate specificity, whereas Clr3 (class II) displayed its main activity on acetylated lysine 14 of histone H3. Thus, the distinct functions of the S. pombe HDACs are likely explained by their distinct cellular localization and their different in vivo specificities. PMID- 11884605 TI - Maintenance of double-stranded telomeric repeats as the critical determinant for cell viability in yeast cells lacking Ku. AB - The Saccharomyces cerevisiae Ku complex, while important for nonhomologous DNA end joining, is also necessary for maintaining wild-type telomere length and a normal chromosomal DNA end structure. Yeast cells lacking Ku can grow at 23 degrees C but are unable to do so at elevated temperatures due to an activation of DNA damage checkpoints. To gain insights into the mechanisms affected by temperature in such strains, we isolated and characterized a new allele of the YKU70 gene, yku70-30(ts). By several criteria, the Yku70-30p protein is functional at 23 degrees C and nonfunctional at 37 degrees C. The analyses of telomeric repeat maintenance as well as the terminal DNA end structure in strains harboring this allele alone or in strains with a combination of other mutations affecting telomere maintenance show that the altered DNA end structure in yeast cells lacking Ku is not generated in a telomerase-dependent fashion. Moreover, the single-stranded G-rich DNA on such telomeres is not detected by DNA damage checkpoints to arrest cell growth, provided that there are sufficient double stranded telomeric repeats present. The results also demonstrate that mutations in genes negatively affecting G-strand synthesis (e.g., RIF1) or C-strand synthesis (e.g., the DNA polymerase alpha gene) allow for the maintenance of longer telomeric repeat tracts in cells lacking Ku. Finally, extending telomeric repeat tracts in such cells at least temporarily suppresses checkpoint activation and growth defects at higher temperatures. Thus, we hypothesize that an aspect of the coordinated synthesis of double-stranded telomeric repeats is sensitive to elevated temperatures. PMID- 11884606 TI - Calcium-dependent assembly of centrin-G-protein complex in photoreceptor cells. AB - Photoexcitation of rhodopsin activates a heterotrimeric G-protein cascade leading to cyclic GMP hydrolysis in vertebrate photoreceptors. Light-induced exchanges of the visual G-protein transducin between the outer and inner segment of rod photoreceptors occur through the narrow connecting cilium. Here we demonstrate that transducin colocalizes with the Ca(2+)-binding protein centrin 1 in a specific domain of this cilium. Coimmunoprecipitation, centrifugation, centrin overlay, size exclusion chromatography, and kinetic light-scattering experiments indicate that Ca(2+)-activated centrin 1 binds with high affinity and specificity to transducin. The assembly of centrin-G-protein complex is mediated by the betagamma-complex. The Ca(2+)-dependent assembly of a G protein with centrin is a novel aspect of the supply of signaling proteins in sensory cells and a potential link between molecular translocations and signal transduction in general. PMID- 11884607 TI - ErbB2/Neu-induced, cyclin D1-dependent transformation is accelerated in p27 haploinsufficient mammary epithelial cells but impaired in p27-null cells. AB - ErbB2/Neu destabilizes the cyclin-dependent kinase (Cdk) inhibitor p27 and increases expression of cyclin D1. Therefore, we studied the roles of p27 and cyclin D1 in ErbB2-mediated mammary epithelial cell transformation. Overexpression of ErbB2 or cyclin D1 in p27(+/-) primary murine mammary epithelial cells resulted in increased proliferation, cyclin D1 nuclear localization, and colony formation in soft agar compared to those in p27(+/+) cells. In contrast, ErbB2- or cyclin D1-overexpressing p27(-/-) cells displayed reduced proliferation, anchorage-independent growth, Cdk4 activity, cyclin D1 expression, and cyclin D1 nuclear localization compared to wild-type cells. A cyclin D1 mutation in its nuclear export sequence (T286A) partially rescued nuclear localization of cyclin D1 in p27(-/-) cells but did not increase proliferation or Cdk4 kinase activity. Overexpression of E2F1, however, increased proliferation to the same degree in p27(+/+), p27(+/-), and p27(-/-) cells. Mammary glands from MMTV (mouse mammary tumor virus)-neu/p27(+/-) mice exhibited alveolar hyperplasia, enhanced proliferation, decreased apoptosis, and accelerated tumor formation compared to MMTV-neu/p27(+/+) glands. However, MMTV neu/p27(-/-) glands showed decreased proliferation, cyclin D1 expression, and Cdk4 activity, as well as markedly prolonged tumor latency, compared to MMTV neu/p27(+/+) glands. These results suggest that p27(+/-) mammary epithelium may be more susceptible to oncogene-induced tumorigenesis, whereas p27-null glands, due to severely impaired cyclin D1/Cdk4 function, are more resistant to transformation. PMID- 11884608 TI - p53-dependent S-phase damage checkpoint and pronuclear cross talk in mouse zygotes with X-irradiated sperm. AB - One difficulty in analyzing the damage response is that the effect of damage itself and that of cellular response are hard to distinguish in irradiated cells. In mouse zygotes, damage can be introduced by irradiated sperm, while damage response can be studied in the unirradiated maternal pronucleus. We have analyzed the p53-dependent damage responses in irradiated-sperm mouse zygotes and found that a p53-responsive reporter was efficiently activated in the female pronucleus. [(3)H]thymidine labeling experiments indicated that irradiated-sperm zygotes were devoid of G(1)/S arrest, but pronuclear DNA synthesis was suppressed equally in male and female pronuclei. p53(-/-) zygotes lacked this suppression, which was corrected by microinjection of glutathione S-transferase-p53 fusion protein. In contrast, p21(-/-) zygotes exhibited the same level of suppression upon fertilization by irradiated sperm. About a half of the 6-Gy-irradiated-sperm zygotes managed to synthesize a full DNA content by prolonging S phase, while the other half failed to do so. Regardless of the DNA content, all the zygotes cleaved to become two-cell-stage embryos. These results revealed the presence of p53-dependent pronuclear cross talk and a novel function of p53 in the S-phase DNA damage checkpoint of mouse zygotes. PMID- 11884609 TI - CENP-A, -B, and -C chromatin complex that contains the I-type alpha-satellite array constitutes the prekinetochore in HeLa cells. AB - CENP-A is a component of centromeric chromatin and defines active centromere regions by forming centromere-specific nucleosomes. We have isolated centromeric chromatin containing the CENP-A nucleosome, CENP-B, and CENP-C from HeLa cells using anti-CENP-A and/or anti-CENP-C antibodies and shown that the CENP-A/B/C complex is predominantly formed on alpha-satellite DNA that contains the CENP-B box (alphaI-type array). Mapping of hypersensitive sites for micrococcal nuclease (MNase) digestion indicated that CENP-A nucleosomes were phased on the alphaI type array as a result of interactions between CENP-B and CENP-B boxes, implying a repetitive configuration for the CENP-B/CENP-A nucleosome complex. Molecular mass analysis by glycerol gradient sedimentation showed that MNase digestion released a CENP-A/B/C chromatin complex of three to four nucleosomes into the soluble fraction, suggesting that CENP-C is a component of the repetitive CENP B/CENP-A nucleosome complex. Quantitative analysis by immunodepletion of CENP-A nucleosomes showed that most of the CENP-C and approximately half the CENP-B took part in formation of the CENP-A/B/C chromatin complex. A kinetic study of the solubilization of CENPs showed that MNase digestion first released the CENP-A/B/C chromatin complex into the soluble fraction, and later removed CENP-B and CENP-C from the complex. This result suggests that CENP-A nucleosomes form a complex with CENP-B and CENP-C through interaction with DNA. On the basis of these results, we propose that the CENP-A/B/C chromatin complex is selectively formed on the I-type alpha-satellite array and constitutes the prekinetochore in HeLa cells. PMID- 11884610 TI - Reversal of growth suppression by p107 via direct phosphorylation by cyclin D1/cyclin-dependent kinase 4. AB - p107 functions to control cell division and development through interaction with members of the E2F family of transcription factors. p107 is phosphorylated in a cell cycle-regulated manner, and its phosphorylation leads to its release from E2F. Although it is known that p107 physically associates with E- and A-type cyclin/cyclin-dependent kinase 2 (Cdk2) complexes through a cyclin-binding RXL motif located in the spacer domain, the mechanisms underlying p107 inactivation via phosphorylation remain poorly defined. Recent genetic evidence indicates a requirement for cyclin D1/Cdk4 complexes in p107 inactivation. In this work, we provide direct biochemical evidence for the involvement of cyclin D1/Cdk4 in the inactivation of p107's growth-suppressive function. While coexpression of cyclin D1/Cdk4 can reverse the cell cycle arrest properties of p107 in Saos-2 cells, we find that p107 in which the Lys-Arg-Arg-Leu sequence of the RXL motif is replaced by four alanine residues is largely refractory to inactivation by cyclin D/Cdk4, indicating a role for this motif in p107 inactivation without a requirement for its tight interaction with cyclin D1/Cdk4. We identified four phosphorylation sites in p107 (Thr-369, Ser-640, Ser-964, and Ser-975) that are efficiently phosphorylated by Cdk4 but not by Cdk2 in vitro and are also phosphorylated in tissue culture cells. Growth suppression by p107 containing nonphosphorylatable residues in these four sites is not reversed by coexpression of cyclin D1/Cdk4. In model p107 spacer region peptides, phosphorylation of S640 by cyclin D1/Cdk4 is strictly dependent upon an intact RXL motif, but phosphorylation of this site in the absence of an RXL motif can be partially restored by replacement of S643 by arginine. This suggests that one role for the RXL motif is to facilitate phosphorylation of nonconsensus Cdk substrates. Taken together, these data indicate that p107 is inactivated by cyclin D1/Cdk4 via direct phosphorylation and that the RXL motif of p107 plays a role in its inactivation by Cdk4 in the absence of stable binding. PMID- 11884611 TI - hnRNP A1 nucleocytoplasmic shuttling activity is required for normal myelopoiesis and BCR/ABL leukemogenesis. AB - hnRNP A1 is a nucleocytoplasmic shuttling heterogeneous nuclear ribonucleoprotein that accompanies eukaryotic mRNAs from the active site of transcription to that of translation. Although the importance of hnRNP A1 as a regulator of nuclear pre mRNA and mRNA processing and export is well established, it is unknown whether this is relevant for the control of proliferation, survival, and differentiation of normal and transformed cells. We show here that hnRNP A1 levels are increased in myeloid progenitor cells expressing the p210(BCR/ABL) oncoprotein, in mononuclear cells from chronic myelogenous leukemia (CML) blast crisis patients, and during disease progression. In addition, in myeloid progenitor 32Dcl3 cells, BCR/ABL stabilizes hnRNP A1 by preventing its ubiquitin/proteasome-dependent degradation. To assess the potential role of hnRNP A1 nucleocytoplasmic shuttling activity in normal and leukemic myelopoiesis, a mutant defective in nuclear export was ectopically expressed in parental and BCR/ABL-transformed myeloid precursor 32Dcl3 cells, in normal murine marrow cells, and in mononuclear cells from a CML patient in accelerated phase. In normal cells, expression of this mutant enhanced the susceptibility to apoptosis induced by interleukin-3 deprivation, suppressed granulocytic differentiation, and induced massive cell death of granulocyte colony-stimulating factor-treated cultures. In BCR/ABL transformed cells, its expression was associated with suppression of colony formation and reduced tumorigenic potential in vivo. Moreover, interference with hnRNP A1 shuttling activity resulted in downmodulation of C/EBPalpha, the major regulator of granulocytic differentiation, and Bcl-X(L), an important survival factor for hematopoietic cells. Together, these results suggest that the shuttling activity of hnRNP A1 is important for the nucleocytoplasmic trafficking of mRNAs that encode proteins influencing the phenotype of normal and BCR/ABL transformed myeloid progenitors. PMID- 11884612 TI - Developmental control of histone mRNA and dSLBP synthesis during Drosophila embryogenesis and the role of dSLBP in histone mRNA 3' end processing in vivo. AB - In metazoans, the 3' end of histone mRNA is not polyadenylated but instead ends with a stem-loop structure that is required for cell cycle-regulated expression. The sequence of the stem-loop in the Drosophila melanogaster histone H2b, H3, and H4 genes is identical to the consensus sequence of other metazoan histone mRNAs, but the sequence of the stem-loop in the D. melanogaster histone H2a and H1 genes is novel. dSLBP binds to these novel stem-loop sequences as well as the canonical stem-loop with similar affinity. Eggs derived from females containing a viable, hypomorphic mutation in dSLBP store greatly reduced amounts of all five histone mRNAs in the egg, indicating that dSLBP is required in the maternal germ line for production of each histone mRNA. Embryos deficient in zygotic dSLBP function accumulate poly(A)(+) versions of all five histone mRNAs as a result of usage of polyadenylation signals located 3' of the stem-loop in each histone gene. Since the 3' ends of adjacent histone genes are close together, these polyadenylation signals may ensure the termination of transcription in order to prevent read through into the next gene, which could possibly disrupt transcription or produce antisense histone mRNA that might trigger RNA interference. During early wild type embryogenesis, ubiquitous zygotic histone gene transcription is activated at the end of the syncytial nuclear cycles during S phase of cycle 14, silenced during the subsequent G(2) phase, and then reactivated near the end of that G(2) phase in the well-described mitotic domain pattern. There is little or no dSLBP protein provided maternally in wild-type embryos, and zygotic expression of dSLBP is immediately required to process newly made histone pre-mRNA. PMID- 11884613 TI - Identification of a novel hypoxia-inducible factor 1-responsive gene, RTP801, involved in apoptosis. AB - Hypoxia is an important factor that elicits numerous physiological and pathological responses. One of the major gene expression programs triggered by hypoxia is mediated through hypoxia-responsive transcription factor hypoxia inducible factor 1 (HIF-1). Here, we report the identification and cloning of a novel HIF-1-responsive gene, designated RTP801. Its strong up-regulation by hypoxia was detected both in vitro and in vivo in an animal model of ischemic stroke. When induced from a tetracycline-repressible promoter, RTP801 protected MCF7 and PC12 cells from hypoxia in glucose-free medium and from H(2)O(2) triggered apoptosis via a dramatic reduction in the generation of reactive oxygen species. However, expression of RTP801 appeared toxic for nondividing neuron-like PC12 cells and increased their sensitivity to ischemic injury and oxidative stress. Liposomal delivery of RTP801 cDNA to mouse lungs also resulted in massive cell death. Thus, the biological effect of RTP801 overexpression depends on the cell context and may be either protecting or detrimental for cells under conditions of oxidative or ischemic stresses. Altogether, the data suggest a complex type of involvement of RTP801 in the pathogenesis of ischemic diseases. PMID- 11884614 TI - The ubiquitin ligase component Siah1a is required for completion of meiosis I in male mice. AB - The mammalian Siah genes encode highly conserved proteins containing a RING domain. As components of E3 ubiquitin ligase complexes, Siah proteins facilitate the ubiquitination and degradation of diverse protein partners including beta catenin, N-CoR, and DCC. We used gene targeting in mice to analyze the function of Siah1a during mammalian development and reveal novel roles in growth, viability, and fertility. Mutant animals have normal weights at term but are postnatally growth retarded, despite normal levels of pituitary growth hormone. Embryonic fibroblasts isolated from mutant animals grow normally. Most animals die before weaning, and few survive beyond 3 months. Serum gonadotropin levels are normal in Siah1a mutant mice; however, females are subfertile and males are sterile due to a block in spermatogenesis. Although spermatocytes in mutant mice display normal meiotic prophase and meiosis I spindle formation, they accumulate at metaphase to telophase of meiosis I and subsequently undergo apoptosis. The requirement of Siah1a for normal progression beyond metaphase I suggests that Siah1a may be part of a novel E3 complex acting late in the first meiotic division. PMID- 11884615 TI - Opposing roles of the extracellular signal-regulated kinase and p38 mitogen activated protein kinase cascades in Ras-mediated downregulation of tropomyosin. AB - We showed previously that activated Ras, but not Raf, causes transformation of RIE-1 epithelial cells, demonstrating the importance of Raf-independent pathways in mediating Ras transformation. To assess the mechanism by which Raf-independent effector signaling pathways contribute to Ras-mediated transformation, we recently utilized representational difference analysis to identify genes expressed in a deregulated fashion by activated Ras but not Raf. One gene identified in these analyses encodes for alpha-tropomyosin. Therefore, we evaluated the mechanism by which Ras causes the downregulation of tropomyosin expression. By using RIE-1 cells that harbor inducible expression of activated H Ras(12V), we determined that the downregulation of tropomyosin expression correlated with the onset of morphological transformation. We found that the reversal of Ras transformation caused by inhibition of extracellular signal regulated kinase activation corresponded to a restoration of tropomyosin expression. Inhibition of p38 activity in Raf-expressing RIE-1 cells caused both morphological transformation and loss of tropomyosin expression. Thus, a reduction in tropomyosin expression correlated strictly with morphological transformation of RIE-1 cells. However, forced overexpression of tropomyosin in Ras-transformed cells did not reverse morphological or growth transformation, a finding consistent with the possibility that multiple changes in gene expression contribute to Ras transformation. We also determined that tropomyosin expression was low in two human tumor cell lines, DLD-1 and HT1080, that harbor endogenous mutated alleles of ras, but high in transformation-impaired, derivative cell lines in which the mutant ras allele has been genetically deleted. Finally, treatment with azadeoxycytidine restored tropomyosin expression in Ras transformed RIE-1, HT1080, and DLD-1 cells, suggesting a role for DNA methylation in downregulating tropomyosin expression. PMID- 11884617 TI - Caveolin-2-deficient mice show evidence of severe pulmonary dysfunction without disruption of caveolae. AB - Caveolin-2 is a member of the caveolin gene family with no known function. Although caveolin-2 is coexpressed and heterooligomerizes with caveolin-1 in many cell types (most notably adipocytes and endothelial cells), caveolin-2 has traditionally been considered the dispensable structural partner of the widely studied caveolin-1. We now directly address the functional significance of caveolin-2 by genetically targeting the caveolin-2 locus (Cav-2) in mice. In the absence of caveolin-2 protein expression, caveolae still form and caveolin-1 maintains its localization in plasma membrane caveolae, although in certain tissues caveolin-1 is partially destabilized and shows modestly diminished protein levels. Despite an intact caveolar membrane system, the Cav-2-null lung parenchyma shows hypercellularity, with thickened alveolar septa and an increase in the number of endothelial cells. As a result of these pathological changes, these Cav-2-null mice are markedly exercise intolerant. Interestingly, these Cav 2-null phenotypes are identical to the ones we and others have recently reported for Cav-1-null mice. As caveolin-2 expression is also severely reduced in Cav-1 null mice, we conclude that caveolin-2 deficiency is the clear culprit in this lung disorder. Our analysis of several different phenotypes observed in caveolin 1-deficient mice (i.e., abnormal vascular responses and altered lipid homeostasis) reveals that Cav-2-null mice do not show any of these other phenotypes, indicating a selective role for caveolin-2 in lung function. Taken together, our data show for the first time a specific role for caveolin-2 in mammalian physiology independent of caveolin-1. PMID- 11884616 TI - Targeted mutagenesis of the Hira gene results in gastrulation defects and patterning abnormalities of mesoendodermal derivatives prior to early embryonic lethality. AB - The Hira gene encodes a nuclear WD40 domain protein homologous to the yeast transcriptional corepressors Hir1p and Hir2p. Using targeted mutagenesis we demonstrate that Hira is essential for murine embryogenesis. Analysis of inbred 129Sv embryos carrying the null mutation revealed an initial requirement during gastrulation, with many mutant embryos having a distorted primitive streak. Mutant embryos recovered at later stages have a range of malformations with axial and paraxial mesendoderm being particularly affected, a finding consistent with the disruption of gastrulation seen earlier in development. This phenotype could be partially rescued by a CD1 genetic background, although the homozygous mutation was always lethal by embryonic day 11, with death probably resulting from abnormal placentation and failure of cardiac morphogenesis. PMID- 11884618 TI - RACK1, an insulin-like growth factor I (IGF-I) receptor-interacting protein, modulates IGF-I-dependent integrin signaling and promotes cell spreading and contact with extracellular matrix. AB - The insulin-like growth factor I (IGF-I) receptor (IGF-IR) is known to regulate a variety of cellular processes including cell proliferation, cell survival, cell differentiation, and cell transformation. IRS-1 and Shc, substrates of the IGF IR, are known to mediate IGF-IR signaling pathways such as those of mitogen activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K), which are believed to play important roles in some of the IGF-IR-dependent biological functions. We used the cytoplasmic domain of IGF-IR in a yeast two-hybrid interaction trap to identify IGF-IR-interacting molecules that may potentially mediate IGF-IR-regulated functions. We identified RACK1, a WD repeat family member and a Gbeta homologue, and demonstrated that RACK1 interacts with the IGF IR but not with the closely related insulin receptor (IR). In several types of mammalian cells, RACK1 interacted with IGF-IR, protein kinase C, and beta1 integrin in response to IGF-I and phorbol 12-myristate 13-acetate stimulation. Whereas most of RACK1 resides in the cytoskeletal compartment of the cytoplasm, transformation of fibroblasts and epithelial cells by v-Src, oncogenic IR or oncogenic IGF-IR, but not by Ros or Ras, resulted in a significantly increased association of RACK1 with the membrane. We examined the role of RACK1 in IGF-IR mediated functions by stably overexpressing RACK1 in NIH 3T3 cells that expressed an elevated level of IGF-IR. RACK1 overexpression resulted in reduced IGF-I induced cell growth in both anchorage-dependent and anchorage-independent conditions. Overexpression of RACK1 also led to enhanced cell spreading, increased stress fibers, and increased focal adhesions, which were accompanied by increased tyrosine phosphorylation of focal adhesion kinase and paxillin. While IGF-I-induced activation of IRS-1, Shc, PI3K, and MAPK pathways was unaffected, IGF-I-inducible beta1 integrin-associated kinase activity and association of Crk with p130(CAS) were significantly inhibited by RACK1 overexpression. In RACK1 overexpressing cells, delayed cell cycle progression in G(1) or G(1)/S was correlated with retinoblastoma protein hypophophorylation, increased levels of p21(Cip1/WAF1) and p27(Kip1), and reduced IGF-I-inducible Cdk2 activity. Reduction of RACK1 protein expression by antisense oligonucleotides prevented cell spreading and suppressed IGF-I-dependent monolayer growth. Our data suggest that RACK1 is a novel IGF-IR signaling molecule that functions as a positive mediator of cell spreading and contact with extracellular matrix, possibly through a novel IGF-IR signaling pathway involving integrin and focal adhesion signaling molecules. PMID- 11884619 TI - Essential regions of Saccharomyces cerevisiae telomerase RNA: separate elements for Est1p and Est2p interaction. AB - The Saccharomyces cerevisiae telomerase RNA subunit is encoded by the TLC1 gene. A selection for viable alleles of TLC1 RNA from a large library of random deletion alleles revealed that less than half (approximately 0.5 kb of the approximately 1.3-kb RNA) is required for telomerase function in vivo. The main essential region (430 nucleotides), which contains the template for telomeric DNA synthesis, was required for coimmunoprecipitation with Est1p and Est2p. Furthermore, the subregion required for interaction with Est1p, the telomerase recruitment subunit, differed from those required for interaction with Est2p, the reverse transcriptase subunit. Two regions of the RNA distant from the template in the nucleotide sequence were required for Est2p binding, but the template itself was not. Having the RNA secured to the protein away from the template is proposed to facilitate the translocation of the RNA template through the active site. More generally, our results support a role for the telomerase RNA serving as a scaffold for binding key protein subunits. PMID- 11884621 TI - hMutSbeta is required for the recognition and uncoupling of psoralen interstrand cross-links in vitro. AB - The removal of interstrand cross-links (ICLs) from DNA in higher eucaryotes is not well understood. Here, we show that processing of psoralen ICLs in mammalian cell extracts is dependent upon the mismatch repair complex hMutSbeta but is not dependent upon the hMutSalpha complex or hMlh1. The processing of psoralen ICLs is also dependent upon the nucleotide excision repair proteins Ercc1 and Xpf but not upon other components of the excision stage of this pathway or upon Fanconi anemia proteins. Products formed during the in vitro reaction indicated that the ICL has been removed or uncoupled from the cross-linked substrate in the mammalian cell extracts. Finally, the hMutSbeta complex is shown to specifically bind to psoralen ICLs, and this binding is stimulated by the addition of PCNA. Thus, a novel pathway for processing ICLs has been identified in mammalian cells which involves components of the mismatch repair and nucleotide excision repair pathways. PMID- 11884620 TI - Protein phosphatase 2A forms a molecular complex with Shc and regulates Shc tyrosine phosphorylation and downstream mitogenic signaling. AB - Protein phosphatase 2A (PP2A) is a multimeric serine/threonine phosphatase that carries out multiple functions. Although numerous observations suggest that PP2A plays a major role in downregulation of the mitogen-activated protein (MAP) kinase pathway, the precise mechanisms are unknown. To clarify the role of PP2A in growth factor (insulin, epidermal growth factor [EGF], and insulin-like growth factor 1 [IGF-1]) stimulation of the Ras/MAP kinase pathway, simian virus 40 small t antigen was expressed in Rat-1 fibroblasts which overexpress insulin receptors. Small t antigen is known to specifically inhibit PP2A by binding to the A PP2A regulatory subunit, interfering with the ability of PP2A to bind to its cellular substrates. Overexpressed small t protein was coimmunoprecipitated with PP2A and inhibited cellular PP2A activity but did not inhibit protein phosphatase 1 (PP1) activity. Insulin, IGF-1, and EGF stimulation also inhibited PP2A activity. Growth factor-stimulated Ras, Raf-1, MAP kinase, and mitogen activated extracellular-signal-regulated kinase kinase (MEK) activities were elevated in small-t-antigen-expressing cells. Furthermore, Shc tyrosine phosphorylation and its association with Grb2 were also elevated in small-t antigen-expressing cells. Expression levels of Shc, Ras, MEK, or MAP kinase and phosphorylation of insulin, EGF, and IGF-1 receptors were not altered. Interestingly, we found that PP2A associated with Shc in the basal state and dissociated in response to insulin and EGF and that this dissociation was inhibited by 65% in small-t-antigen-expressing cells. In addition, we found that PP2A associates with the phosphotyrosine-binding domain (PTB domain) of Shc and that phosphorylation of tyrosine 317 of Shc was required for PP2A-Shc dissociation. We conclude (i) that PP2A negatively regulates the Ras/MAP kinase pathway by binding to Shc, inhibiting tyrosine phosphorylation; (ii) that the Shc PP2A association is mediated by the Shc PTB domain but the interaction is independent of phosphotyrosine binding, indicating a new molecular function for the PTB domain; (iii) that growth factor stimulation, or small-t-antigen expression, causes dissociation of the PP2A-Shc complex, facilitating Shc phosphorylation and downstream activations of the Ras/MAP kinase pathway; and (iv) that this defines a new mechanism of small-t-antigen action to promote mitogenesis. PMID- 11884622 TI - Proteolytic cleavage of cyclin E leads to inactivation of associated kinase activity and amplification of apoptosis in hematopoietic cells. AB - Cyclin E/Cdk2 is a critical regulator of cell cycle progression from G(1) to S in mammalian cells and has an established role in oncogenesis. Here we examined the role of deregulated cyclin E expression in apoptosis. The levels of p50-cyclin E initially increased, and this was followed by a decrease starting at 8 h after treatment with genotoxic stress agents, such as ionizing radiation. This pattern was mirrored by the cyclin E-Cdk2-associated kinase activity and a time-dependent expression of a novel p18-cyclin E. p18-cyclin E was induced during apoptosis triggered by multiple genotoxic stress agents in all hematopoietic tumor cell lines we have examined. The p18-cyclin E expression was prevented by Bcl-2 overexpression and by the general caspase and specific caspase 3 pharmacologic inhibitors zVAD-fluoromethyl ketone (zVAD-fmk) and N-acetyl-Asp-Glu-Val-Asp aldehyde (DEVD-CHO), indicating that it was linked to apoptosis. A p18-cyclin E(276-395) (where cyclin E(276-395) is the cyclin E fragment containing residues 276 to 395) was reconstituted in vitro, with mutagenesis experiments, indicating that the caspase-dependent cleavage was at amino acid residues 272 to 275. Immunoprecipitation analyses of the ectopically expressed cyclin E(1-275), cyclin E(276-395) deletion mutants, and native p50-cyclin E demonstrated that caspase mediated cyclin E cleavage eliminated interaction with Cdk2 and therefore inactivated the associated kinase activity. Overexpression of cyclin E(276-395), but not of several other cyclin E mutants, specifically induced phosphatidylserine exposure and caspase activation in a dose-dependent manner, which were inhibited in Bcl-2-overexpressing cells or in the presence of zVAD fmk. Apoptosis and generation of p18-cyclin E were significantly inhibited by overexpressing the cleavage-resistant cyclin E mutant, indicating a functional role for caspase-dependent proteolysis of cyclin E for apoptosis of hematopoietic tumor cells. PMID- 11884624 TI - Requirement of RAD5 and MMS2 for postreplication repair of UV-damaged DNA in Saccharomyces cerevisiae. AB - UV lesions in the template strand block the DNA replication machinery. Genetic studies of the yeast Saccharomyces cerevisiae have indicated the requirement of the Rad6-Rad18 complex, which contains ubiquitin-conjugating and DNA-binding activities, in the error-free and mutagenic modes of damage bypass. Here, we examine the contributions of the REV3, RAD30, RAD5, and MMS2 genes, all of which belong to the RAD6 epistasis group, to the postreplication repair of UV-damaged DNA. Discontinuities, which are formed in DNA strands synthesized from UV-damaged templates, are not repaired in the rad5Delta and mms2Delta mutants, thus indicating the requirement of the Rad5 protein and the Mms2-Ubc13 ubiquitin conjugating enzyme complex in this repair process. Some discontinuities accumulate in the absence of RAD30-encoded DNA polymerase eta (Poleta) but not in the absence of REV3-encoded DNA Polzeta. We concluded that replication through UV lesions in yeast is mediated by at least three separate Rad6-Rad18-dependent pathways, which include mutagenic translesion synthesis by Polzeta, error-free translesion synthesis by Poleta, and postreplication repair of discontinuities by a Rad5-dependent pathway. We suggest that newly synthesized DNA possessing discontinuities is restored to full size by a "copy choice" type of DNA synthesis which requires Rad5, a DNA-dependent ATPase, and also PCNA and Poldelta. The possible roles of the Rad6-Rad18 and the Mms2-Ubc13 enzyme complexes in Rad5 dependent damage bypass are discussed. PMID- 11884623 TI - Base excision repair is limited by different proteins in male germ cell nuclear extracts prepared from young and old mice. AB - The combined observations of elevated DNA repair gene expression, high uracil-DNA glycosylase-initiated base excision repair, and a low spontaneous mutant frequency for a lacI transgene in spermatogenic cells from young mice suggest that base excision repair activity is high in spermatogenic cell types. Notably, the spontaneous mutant frequency of the lacI transgene is greater in spermatogenic cells obtained from old mice, suggesting that germ line DNA repair activity may decline with age. A paternal age effect in spermatogenic cells is recognized for the human population as well. To determine if male germ cell base excision repair activity changes with age, uracil-DNA glycosylase-initiated base excision repair activity was measured in mixed germ cell (i.e., all spermatogenic cell types in adult testis) nuclear extracts prepared from young, middle-aged, and old mice. Base excision repair activity was also assessed in nuclear extracts from premeiotic, meiotic, and postmeiotic spermatogenic cell types obtained from young mice. Mixed germ cell nuclear extracts exhibited an age-related decrease in base excision repair activity that was restored by addition of apurinic/apyrimidinic (AP) endonuclease. Uracil-DNA glycosylase and DNA ligase were determined to be limiting in mixed germ cell nuclear extracts prepared from young animals. Base excision repair activity was only modestly elevated in pachytene spermatocytes and round spermatids relative to other spermatogenic cells. Thus, germ line short-patch base excision repair activity appears to be relatively constant throughout spermatogenesis in young animals, limited by uracil-DNA glycosylase and DNA ligase in young animals, and limited by AP endonuclease in old animals. PMID- 11884625 TI - Comparative analysis of hairpin ribozyme structures and interference data. AB - Great strides in understanding the molecular underpinnings of RNA catalysis have been achieved with advances in RNA structure determination by NMR spectroscopy and X-ray crystallography. Despite these successes the functional relevance of a given structure can only be assessed upon comparison with biochemical studies performed on functioning RNA molecules. The hairpin ribozyme presents an excellent case study for such a comparison. The active site is comprised of two stems each with an internal loop that forms a series of non-canonical base pairs. These loops dock into each other to create an active site for catalysis. Recently, three independent structures have been determined for this catalytic RNA, including two NMR structures of the isolated loop A and loop B stems and a high-resolution crystal structure of both loops in a docked conformation. These structures differ significantly both in their tertiary fold and the nature of the non-canonical base pairs formed within each loop. Several of the chemical groups required to achieve a functioning hairpin ribozyme have been determined by nucleotide analog interference mapping (NAIM). Here we compare the three hairpin structures with previously published NAIM data to assess the convergence between the structural and functional data. While there is significant disparity between the interference data and the individual NMR loop structures, there is almost complete congruity with the X-ray structure. The only significant differences cluster around an occluded pocket adjacent to the scissile phosphate. These local differences may suggest a role for these atoms in the transition state, either directly in chemistry or via a local structural rearrangement. PMID- 11884627 TI - The CUP1 upstream repeated element renders CUP1 promoter activation insensitive to mutations in the RNA polymerase II transcription complex. AB - Activation of transcription in eukaryotes requires the concerted action of numerous components of the RNA polymerase II transcriptional apparatus. The degree of dependence on many of these components varies from gene to gene and it is still largely unknown how the requirement for any particular component is determined at any given gene. We show that removal of Gal11 from the yeast transcription complex can affect activation from the CUP1 UAS in a manner dependent on its genomic context. Our results indicate a novel function for the CUP1 upstream repeated element (CURE) located upstream of the CUP1 UAS at the naturally multimerized CUP1 locus. The presence of CURE endowed the CUP1 UAS with a reduced susceptibility to the effects of deleting Gal11. Similar results were obtained with the Srb/mediator subunit Srb5. Restoration of activation from the CUP1 promoter to wild-type levels by the CURE correlated with changes in the accessibility of local chromatin to nucleases. The CURE sequence may serve to protect the stress-inducible CUP1 UAS-promoter elements against reduced activation that may result from crippled transcription complexes under stress conditions. PMID- 11884628 TI - Differential expression and requirements for Schizosaccharomyces pombe RAD52 homologs in DNA repair and recombination. AB - In fission yeast two RAD52 homologs have been identified, rad22A(+) and rad22B(+). Two-hybrid experiments and GST pull-down assays revealed physical interaction between Rad22A and Rad22B, which is dependent on the N-terminal regions. Interaction with Rhp51 is dependent on the C-terminal parts of either protein. Both Rad22A and Rad22B also interact with RPA. The expression of rad22B(+) in mitotically dividing cells is very low in comparison with rad22A(+) but is strongly enhanced after induction of meiosis, in contrast to rad22A(+). Rad22B mutant cells are not hypersensitive to DNA-damaging agents (X-rays, UV and cisplatin) and display normal levels of recombination. In these respects the Schizosaccharomyces pombe rad22B mutant resembles the weak phenotype of vertebrate cells deficient for RAD52. Mutation of rad22A(+) leads to severe sensitivity to DNA-damaging agents and to defects in recombination. In a rad22Arad22B double mutant a further increase in sensitivity to DNA-damaging agents and additional mitotic recombination defects were observed. The data presented here indicate that Rad22A and Rad22B have overlapping roles in repair and recombination, although specialized functions for each protein cannot be excluded. PMID- 11884626 TI - Real-time PCR in virology. AB - The use of the polymerase chain reaction (PCR) in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting nucleic acids from a number of origins and it has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility and the reduced risk of carry-over contamination. There are currently five main chemistries used for the detection of PCR product during real-time PCR. These are the DNA binding fluorophores, the 5' endonuclease, adjacent linear and hairpin oligoprobes and the self-fluorescing amplicons, which are described in detail. We also discuss factors that have restricted the development of multiplex real-time PCR as well as the role of real-time PCR in quantitating nucleic acids. Both amplification hardware and the fluorogenic detection chemistries have evolved rapidly as the understanding of real-time PCR has developed and this review aims to update the scientist on the current state of the art. We describe the background, advantages and limitations of real-time PCR and we review the literature as it applies to virus detection in the routine and research laboratory in order to focus on one of the many areas in which the application of real-time PCR has provided significant methodological benefits and improved patient outcomes. However, the technology discussed has been applied to other areas of microbiology as well as studies of gene expression and genetic disease. PMID- 11884630 TI - Binding of oligonucleotides to a viral hairpin forming RNA triplexes with parallel G*G*C triplets. AB - Infrared and UV spectroscopies have been used to study the assembly of a hairpin nucleotide sequence (nucleotides 3-30) of the 5' non-coding region of the hepatitis C virus RNA (5'-GGCGGGGAUUAUCCCCGCUGUGAGGCGG-3') with a RNA 20mer ligand (5'-CCGCCUCACAAAGGUGGGGU-3') in the presence of magnesium ion and spermidine. The resulting complex involves two helical structural domains: the first one is an intermolecular duplex stem at the bottom of the target hairpin and the second one is a parallel triplex generated by the intramolecular hairpin duplex and the ligand. Infrared spectroscopy shows that N-type sugars are exclusively present in the complex. This is the first case of formation of a RNA parallel triplex with purine motif and shows that this type of targeting RNA strands to viral RNA duplexes can be used as an alternative to antisense oligonucleotides or ribozymes. PMID- 11884629 TI - Involvement of conserved histidine, lysine and tyrosine residues in the mechanism of DNA cleavage by the caspase-3 activated DNase CAD. AB - The caspase-activated DNase (CAD) is involved in DNA degradation during apoptosis. Chemical modification of murine CAD with the lysine-specific reagent 2,4,6-trinitrobenzenesulphonic acid and the tyrosine-specific reagent N acetylimidazole leads to inactivation of the nuclease, indicating that lysine and tyrosine residues are important for DNA cleavage by this enzyme. The presence of DNA or the inhibitor ICAD-L protects the enzyme from modification. Amino acid substitution in murine CAD of lysines and tyrosines conserved in CADs from five different species leads to variants with little if any catalytic activity, but unaltered DNA binding (K155Q, K301Q, K310Q, Y247F), with the exception of Y170F, which retains wild-type activity. Similarly, as observed for the previously characterised H242N, H263N, H308N and H313N variants, the newly introduced His- >Asp/Glu or Arg exchanges lead to variants with <1% of wild-type activity, with two exceptions: H313R shows wild-type activity, and H308D at pH 5.0 exhibits approximately 5% of wild-type activity at this pH. Y170F and H313R produce a specific pattern of fragments, different from wild-type CAD, which degrades DNA non-specifically. The recombinant nuclease variants produced in Escherichia coli were tested for their ability to form nucleolytically active oligomers. They did not show any significant deviation from the wild-type enzyme. Based on these and published data possible roles of the amino acid residues under investigation are discussed. PMID- 11884631 TI - Optimal transfection with the HK polymer depends on its degree of branching and the pH of endocytic vesicles. AB - We have recently reported that liposomes in combination with histidine (HK) containing polymers enhanced the expression of luciferase in transfected cells. In transformed or malignant cell lines, branched HK polymers (combined with liposome carriers) were significantly more effective than the linear HK polymer in stimulating gene expression. In the current study, we found that the linear HK polymer enhanced gene expression in primary cell lines more effectively than the branched polymers. The differences in the optimal carrier (linear versus branched) were not due to initial cellular uptake, size of the complexes or level of gene expression. There was, however, a strong association between the optimal type of HK polymer and the pH of endocytic vesicles (P = 0.0058). By altering the percentage of histidines carrying a positive charge, the endosomal pH of a cell may determine the amount of DNA released from the linear or branched HK polymer. In the two cell lines in which the linear HK was the optimal polymer, the endocytic vesicles were strongly acidic with a pH of <5.0. Conversely, in the four cell lines in which the branched polymers were optimal transfection agents, the pH of endocytic vesicles was >6.0. Furthermore, binding data support the relationship between DNA release from the optimal HK polymer and endosomal pH. The interplay between optimal HK polymers and the endosomal pH may lead to improved gene-delivery polymers tailored to a particular cell. PMID- 11884632 TI - Human Rad54B is a double-stranded DNA-dependent ATPase and has biochemical properties different from its structural homolog in yeast, Tid1/Rdh54. AB - The RAD52 epistasis group genes are involved in homologous recombination, and they are conserved from yeast to humans. We have cloned a novel human gene, RAD54B, which is homologous to yeast and human RAD54. Human Rad54B (hRad54B) shares high homology with human Rad54 (hRad54) in the central region containing the helicase motifs characteristic of the SNF2/SWI2 family of proteins, but the N terminal domain is less conserved. In yeast, another RAD54 homolog, TID1/RDH54, plays a role in recombination. Tid1/Rdh54 interacts with yeast Rad51 and a meiosis-specific Rad51 homolog, Dmc1. The N-terminal domain of hRad54B shares homology with that of Tid1/Rdh54, suggesting that Rad54B may be the human counterpart of Tid1/Rdh54. We purified the hRad54 and hRad54B proteins from baculovirus-infected insect cells and examined their biochemical properties. hRad54B, like hRad54, is a DNA-binding protein and hydrolyzes ATP in the presence of double-stranded DNA, though its rate of ATP hydrolysis is lower than that of hRad54. Human Rad51 interacts with hRad54 and enhances its ATPase activity. In contrast, neither human Rad51 nor Dmc1 directly interacts with hRad54B. Although hRad54B is the putative counterpart of Tid1/Rdh54, our findings suggest that hRad54B behaves differently from Tid1/Rdh54. PMID- 11884633 TI - Heat-induced formation of reactive oxygen species and 8-oxoguanine, a biomarker of damage to DNA. AB - Heat-induced formation of 8-oxoguanine was demonstrated in DNA solutions in 10( 3) M phosphate buffer, pH 6.8, by enzyme-linked immunosorbent assays using monoclonal antibodies against 8-oxoguanine. A radiation-chemical yield of 3.7 x 10(-2) micromol x J(-1) for 8-oxoguanine production in DNA upon gamma-irradiation was used as an adequate standard for quantitation of 8-oxoguanine in whole DNA. The initial yield of heat-induced 8-oxoguanine exhibits first order kinetics. The rate constants for 8-oxoguanine formation were determined at elevated temperatures; the activation energy was found to be 27 +/- 2 kcal/mol. Extrapolation to 37 degrees C gave a value of k37 = 4.7 x 10(-10) x s(-1). Heat induced 8-oxoguanine formation and depurination of guanine and adenine show similarities of the processes, which implies that heat-mediated generation of reactive oxygen species (ROS) should occur. Heat-induced production of H2O2 in phosphate buffer was shown. The sequence of reactions of thermally mediated ROS formation have been established: activation of dissolved oxygen to the singlet state, generation of superoxide radicals and their dismutation to H2O2. Gas saturation (O2, N2 and Ar), D2O, scavengers of 1O2, O2-* and OH* radicals and metal chelators influenced heat-induced 8-oxoguanine formation as they affected thermal ROS generation. These findings imply that heat acts via ROS attack leading to oxidative damage to DNA. PMID- 11884634 TI - Methylation of adenine in the nuclear DNA of Tetrahymena is internucleosomal and independent of histone H1. AB - There are about 50 copies of each chromosome in the somatic macronucleus of the ciliated protozoan TETRAHYMENA: Approximately 0.8% of the adenine residues in the macronuclear DNA of Tetrahymena are methylated to N6-methyladenine. The degree of methylation varies between sites from a very low percentage to >90%. In this study a correlation was found between nucleosome positioning and DNA methylation. Eight GATC sites with different levels of methylation were examined. There was a direct correlation between the degree of methylation and proximity to linker DNA at these sites. Although methylation occurs preferentially in linker DNA, the patterns and extent of methylation in a histone H1 knockout strain were virtually indistinguishable from those in wild-type cells. PMID- 11884635 TI - The solution structure of an oligonucleotide duplex containing a 2' deoxyadenosine-3-(2-hydroxyethyl)- 2'-deoxyuridine base pair determined by NMR and molecular dynamics studies. AB - Determination of the solution structure of the duplex d(GCAAGTC(HE)AAAACG)*d(CGTTTTAGACTTGC) containing a 3-(2-hydroxyethyl)-2' deoxyuridine*deoxyadenine (HE*A) base pair is reported. The three-dimensional solution structure, determined starting from 512 models via restrained molecular mechanics using inter-proton distances and torsion angles, converged to two final families of structures. For both families the HE and the opposite A residues are intrahelical and in the anti conformation. The hydroxyethyl chain lies close to the helix axis and for one family the hydroxyl group is above the HE*A plane and in the other case it is below. These two models were used to start molecular dynamic calculations with explicit solvent to explore the hydrogen bonding possibilities of the HE*A base pair. The dynamics calculations converge finally to one model structure in which two hydrogen bonds are formed. The first is formed all the time and is between HEO4 and the amino group of A, and the second, an intermittent one, is between the hydroxyl group and the N1 of A. When this second hydrogen bond is not formed a weak interaction CH...N is possible between HEC7H2 and N1A21. All the best structures show an increase in the C1'-C1' distance relative to a Watson-Crick base pair. PMID- 11884636 TI - Phi29 DNA polymerase residues Tyr59, His61 and Phe69 of the highly conserved ExoII motif are essential for interaction with the terminal protein. AB - Phage Phi29 encodes a DNA-dependent DNA polymerase belonging to the eukaryotic type (family B) subgroup of DNA polymerases that use a protein as the primer for initiation of DNA synthesis. In one of the most important motifs present in the 3'-->5' exonucleolytic domain of proofreading DNA polymerases, the ExoII motif, Phi29 DNA polymerase contains three amino acid residues, Y59, H61 and F69, which are highly conserved among most proofreading DNA polymerases. These residues have recently been shown to be involved in proper stabilization of the primer terminus at the 3'-->5' exonuclease active site. Here we investigate by means of site directed mutagenesis the role of these three residues in reactions that are specific for DNA polymerases utilizing a protein-primed DNA replication mechanism. Mutations introduced at residues Y59, H61 and F69 severely affected the protein-primed replication capacity of Phi29 DNA polymerase. For four of the mutants, namely Y59L, H61L, H61R and F69S, interaction with the terminal protein was affected, leading to few initiation and transition products. These findings, together with the specific conservation of Y59, H61 and F69 among DNA polymerases belonging to the protein-primed subgroup, strongly suggest a functional role of these amino acid residues in the DNA polymerase-terminal protein interaction. PMID- 11884638 TI - Representational difference analysis in a lupus-prone mouse strain results in the identification of an unstable region of the genome on chromosome 11. AB - BXSB mice develop a lupus-like autoimmune syndrome. We have previously identified several intervals that were linked to disease and, in an attempt to characterise lupus susceptibility genes within these intervals, we have sought to isolate differentially expressed genes. Representational difference analysis was used to compare gene expression between BXSB and C57BL/10 mice using spleen and thymus as a source of mRNA. The majority of differentially expressed sequences identified were immunoglobulin and gp70-related sequences, overexpression of these being characteristic of the disease. Among other isolated sequences were a sialyltransferase gene, a mouse tumour virus superantigen gene (Mtv-3), and the virus-related sequence, hitchhiker. In BXSB the sialyltransferase gene not only overexpressed spliced transcripts, but also produced high levels of unspliced mRNA. Further analysis demonstrated that the copy number of the three linked sequences: sialyltransferase, Mtv-3 and hitchhiker, was amplified in BXSB and that the structural organisation of this locus varies in different mouse strains. This locus consists of three parts, Mtv-3-hitchhiker-sialyltransferase, hitchhiker-sialyltransferase, and sialyltransferase alone. Different combinations of the regions are present in different mouse strains. Linkage analysis demonstrated that this region at the distal end of chromosome 11 is weakly linked to phenotypic markers of disease. PMID- 11884637 TI - Transcriptional regulation of the mouse steroid 5alpha-reductase type II gene by progesterone in brain. AB - The steroid 5alpha-reductase (5alpha-R) plays an important physiological role in the conversion of steroid hormones such as androgen and progesterone to their 5alpha-reduced derivatives. 5alpha-R type II (5alpha-R2), one of two 5alpha-R isoforms, is thought to be a key enzyme in the generation of neuroactive steroids in the brain, particularly allopregnanolone (AP), via the production of its precursor dihydroprogesterone from progesterone. In the present study, we investigated possible regulatory mechanisms of 5alpha-R2 gene expression by steroid hormones in the female mouse brain. We first cloned mouse 5alpha-R2 (m5alpha-R2) cDNA by degenerate PCR, and found that progesterone induced 5alpha R2 gene expression to levels detectable by in situ hybridization in female mouse brains. Functional analysis of the m5alpha-R2 gene promoter by a transient expression assay with human progesterone receptor (PR) and androgen receptor (AR) expression vectors identified a progesterone and androgen regulatory element (m5alpha-R2 PRE/ARE). Results of an electrophoretic mobility shift assay revealed that both PR and AR homodimers bound directly to m5alpha-R2 PRE/ARE sequence. These findings suggest that the gene expression of m5alpha-R2 is transcriptionally regulated by progesterone in female brains. PMID- 11884639 TI - Boron-containing aptamers to ATP. AB - Boron neutron capture therapy (BNCT), an experimental treatment for certain cancers, destroys only cells near the boron; however, there is a need to develop highly specific delivery agents. As nucleic acid aptamers recognize specific molecular targets, we investigated the influence of boronated nucleotide analogs on RNA function and on the systematic evolution of ligands by exponential enrichment (SELEX) process. Substitution of guanosine 5'-(alpha-P-borano) triphosphate (bG) for GTP or uridine 5'-(alpha-P-borano) triphosphate (bU) for UTP in several known aptamers diminished or eliminated target recognition by those RNAs. Specifically, ATP-binding aptamers containing the zeta-fold, which appears in several selections for adenosine aptamers, became inactive upon bG substitution but were only moderately affected by bU substitution. Selections were carried out using the bG or bU analogs with C8-linked ATP agarose as the binding target. The selections with bU and normal NTP yielded some zeta-fold aptamers, while the bG selection yielded none of this type. Non-zeta aptamers from bU and bG populations tolerated the borano substitution and many required it. The borano nucleotide requirement is specific; bU could not be used in bG dependent aptamers nor vice versa. The borano group plays an essential role, as yet undefined, in target recognition or RNA structure. We conclude that the bG and bU nucleotides are fully compatible with SELEX, and that these analogs could be used to make boronated aptamers as therapeutics for BNCT. PMID- 11884640 TI - DAP-like kinase interacts with the rat homolog of Schizosaccharomyces pombe CDC5 protein, a factor involved in pre-mRNA splicing and required for G2/M phase transition. AB - DAP-like kinase (Dlk, also termed ZIP kinase) is a leucine zipper-containing serine/threonine-specific protein kinase with as yet unknown biological function(s). Interaction partners so far identified are either transcription factors or proteins that can support or counteract apoptosis. Thus, Dlk might be involved in regulating transcription or, more generally, survival or apoptosis. Here we report on a new interaction partner, the rat homolog of Schizosaccharomyces pombe CDC5 protein, a presumptive transcription and splicing factor involved in the G(2)/M transition. In vitro, rat CDC5 forms complexes with, but is not phosphorylated by, Dlk. Rather, it was phosphorylated by an associated kinase which was identified as CK2. The interaction domain of Dlk was mapped to the leucine zipper, while that of CDC5 was mapped to the C-terminal region between residues 500 and 802. In vivo, both proteins co-localize perfectly in distinct speckle-like structures in the nucleus, some of which overlap with promyelocytic leukemia protein. Interestingly, splicing factor SC35, which also resides in speckles, was partially displaced upon overexpression of either CDC5 or Dlk, perhaps due to phosphorylation by Dlk. Together with previous data, these results suggest that Dlk might play a role in coordinating specific transcription and splicing events. PMID- 11884641 TI - Mining Bacillus subtilis chromosome heterogeneities using hidden Markov models. AB - We present here the use of a new statistical segmentation method on the Bacillus subtilis chromosome sequence. Maximum likelihood parameter estimation of a hidden Markov model, based on the expectation-maximization algorithm, enables one to segment the DNA sequence according to its local composition. This approach is not based on sliding windows; it enables different compositional classes to be separated without prior knowledge of their content, size and localization. We compared these compositional classes, obtained from the sequence, with the annotated DNA physical map, sequence homologies and repeat regions. The first heterogeneity revealed discriminates between the two coding strands and the non coding regions. Other main heterogeneities arise; some are related to horizontal gene transfer, some to t-enriched composition of hydrophobic protein coding strands, and others to the codon usage fitness of highly expressed genes. Concerning potential and established gene transfers, we found 9 of the 10 known prophages, plus 14 new regions of atypical composition. Some of them are surrounded by repeats, most of their genes have unknown function or possess homology to genes involved in secondary catabolism, metal and antibiotic resistance. Surprisingly, we notice that all of these detected regions are a + t richer than the host genome, raising the question of their remote sources. PMID- 11884642 TI - A second set of loxP marker cassettes for Cre-mediated multiple gene knockouts in budding yeast. AB - Heterologous markers are important tools required for the molecular dissection of gene function in many organisms, including Saccharomyces cerevisiae. Moreover, the presence of gene families and isoenzymes often makes it necessary to delete more than one gene. We recently introduced a new and efficient gene disruption cassette for repeated use in budding yeast, which combines the heterologous dominant kan(r) resistance marker with a Cre/loxP-mediated marker removal procedure. Here we describe an additional set of four completely heterologous loxP-flanked marker cassettes carrying the genes URA3 and LEU2 from Kluyveromyces lactis, his5(+) from Schizosaccharomyces pombe and the dominant resistance marker ble(r) from the bacterial transposon Tn5, which confers resistance to the antibiotic phleomycin. All five loxP--marker gene--loxP gene disruption cassettes can be generated using the same pair of oligonucleotides and all can be used for gene disruption with high efficiency. For marker rescue we have created three additional Cre expression vectors carrying HIS3, TRP1 or ble(r) as the yeast selection marker. The set of disruption cassettes and Cre expression plasmids described here represents a significant further development of the marker rescue system, which is ideally suited to functional analysis of the yeast genome. PMID- 11884643 TI - Construction and electrophoretic migration of single-stranded DNA knots and catenanes. AB - In recent years there has been growing interest in the question of how the particular topology of polymeric chains affects their overall dimensions and physical behavior. The majority of relevant studies are based on numerical simulation methods or analytical treatment; however, both these approaches depend on various assumptions and simplifications. Experimental verification is clearly needed but was hampered by practical difficulties in obtaining preparative amounts of knotted or catenated polymers with predefined topology and precisely set chain length. We introduce here an efficient method of production of various single-stranded DNA knots and catenanes that have the same global chain length. We also characterize electrophoretic migration of the produced single-stranded DNA knots and catenanes with increasing complexity. PMID- 11884644 TI - A rapid, quantitative, non-radioactive bisulfite-SNuPE- IP RP HPLC assay for methylation analysis at specific CpG sites. AB - The precise mapping and quantification of DNA methylation as an epigenetic parameter during development and in diseased tissues is of great importance for functional genomics. Here we describe a rapid, quantitative method to assess methylation levels at specific CpG sites using PCR products of bisulfite-treated genomic DNA. Using single nucleotide primer extension (SNuPE) assays in combination with ion pair reverse phase high performance liquid chromatography (IP RP HPLC) separation techniques, methylated and unmethylated CpGs can be discriminated and quantified based on the different masses and hydrophobicities of the extended primer products. The assay is linear, highly reproducible and several sites can be measured simultaneously in one reaction. It can be semi automated and eliminates the need for cloning and sequencing of individual bisulfite PCR products. PMID- 11884645 TI - Principles of quantitation of viral loads using nucleic acid sequence-based amplification in combination with homogeneous detection using molecular beacons. AB - For quantitative NASBA-based viral load assays using homogeneous detection with molecular beacons, such as the NucliSens EasyQ HIV-1 assay, a quantitation algorithm is required. During the amplification process there is a constant growth in the concentration of amplicons to which the beacon can bind while generating a fluorescence signal. The overall fluorescence curve contains kinetic information on both amplicon formation and beacon binding, but only the former is relevant for quantitation. In the current paper, mathematical modeling of the relevant processes is used to develop an equation describing the fluorescence curve as a function of the amplification time and the relevant kinetic parameters. This equation allows reconstruction of RNA formation, which is characterized by an exponential increase in concentrations as long as the primer concentrations are not rate limiting and by linear growth over time after the primer pool is depleted. During the linear growth phase, the actual quantitation is based on assessing the amplicon formation rate from the viral RNA relative to that from a fixed amount of calibrator RNA. The quantitation procedure has been successfully applied in the NucliSens EasyQ HIV-1 assay. PMID- 11884646 TI - Hierarchical high-throughput SNP genotyping of the human Y chromosome using MALDI TOF mass spectrometry. AB - We have established the use of a primer extension/mass spectrometry method (the PinPoint assay) for high-throughput SNP genotyping of the human Y chromosome. 118 markers were used to define 116 haplogroups and typing was organised in a hierarchical fashion. Twenty multiplex PCR/primer extension reactions were set up and each sample could be assigned to a haplogroup with only two to five of these multiplex analyses. A single aliquot of one enzyme was found to be sufficient for both PCR and primer extension. We observed 100% accuracy in blind validation tests. The technique thus provides a reliable, cost-effective and automated method for Y genotyping, and the advantages of using a hierarchical strategy can be applied to any DNA segment lacking recombination. PMID- 11884648 TI - Deja vu: possible parahippocampal mechanisms. AB - Deja vu experiences are common in normal subjects. In addition, they are established symptoms of temporal lobe seizures. The author argues that the phenomenon is the result of faulty and isolated activity of a recognition memory system that consists of the parahippocampal gyrus and its neocortical connections. This memory system is responsible for judgments of familiarity. The result is that a momentary perceived scene is given the characteristics of familiarity that normally accompany a conscious recollection. The normal functioning of other brain structures involved in memory retrieval--the prefrontal cortex and the hippocampus proper--leads to the perplexing phenomenological quality of deja vu. The hypothesis accounts for many characteristics of deja vu in healthy subjects and is well fitting with experimental findings in patients with epilepsy. PMID- 11884647 TI - The future for diffusion tensor imaging in neuropsychiatry. PMID- 11884649 TI - Estimating the prevalence of agitation in community-dwelling persons with Alzheimer's disease. AB - To estimate the prevalence of, and develop norms for, significant agitation in community-dwelling persons with Alzheimer's disease (AD), the authors applied three different criteria to persons with AD (n=235) and normal elderly control subjects (NEC; n=64). The criteria were used to identify the minimum total score on the Cohen-Mansfield Agitation Inventory (CMAI) that represents significant or "excessive" agitation and to estimate its prevalence. The "ultraliberal" criterion resulted in 99.1% of persons with AD and 56.6% of NEC being classified as "excessively" disturbed. The "liberal" and "conservative" criteria classified 66.7% and 68.2% of persons with AD, and no NEC, as "excessively" disturbed. The authors conclude that the best estimate of prevalence of excessive agitation in this population is 67.5%, and that individuals with CMAI scores of 0 to 14 probably should not be considered to have excessive agitation. PMID- 11884650 TI - Age and regional cerebral blood flow in schizophrenia: age effects in anterior cingulate, frontal, and parietal cortex. AB - Positron emission tomography ([(15)O] water PET) was used to examine the relationship between age and regional cerebral blood flow (rCBF) in schizophrenia. Forty-nine unmedicated male patients, ages 20-51, underwent imaging during an eyes-closed resting condition. Negative correlations were observed between age and rCBF in the anterior cingulate, as well as in frontal (Brodmann area 8) and parietal cortex (area 40) bilaterally. The observation of reduced rCBF in the anterior cingulate with increased age is consistent with previous findings in healthy subjects. In contrast, the reduced flow observed in the frontal and parietal regions may be unique to schizophrenia. PMID- 11884651 TI - Posttraumatic amnesia and recall of a traumatic event following traumatic brain injury. AB - The relationship between posttraumatic amnesia (PTA) and symptoms of posttraumatic stress disorder (PTSD) was examined in 282 outpatients at a mean of 53 days after traumatic brain injury (TBI). Patients were assessed for TBI severity, intrusive and avoidant PTSD-type symptoms, and psychological distress, and were stratified into four comparison groups by duration of PTA. Levels of PTSD-type symptoms and psychological distress did not differ significantly between groups. Even patients with PTA >1 week reported intrusive and avoidant PTSD-type symptoms. However, when patients were stratified into those with PTA of <1 hour or >1 hour, the former were more likely to report such symptoms. TBI patients with brief PTA are more likely to experience PTSD-type reactions, but severe TBI with prolonged PTA is not incompatible with such reactions in a subset of patients. Possible mechanisms that could account for this finding are discussed. PMID- 11884652 TI - Effects of depression and Parkinson's disease on cognitive functioning. AB - This study compared the performance of Parkinson's disease (PD) patients with and without depression, patients with depression alone, and normal control subjects on a cognitive screening instrument, the Mattis Dementia Rating Scale (DRS) to evaluate the influences of depression and Parkinson's disease on cognition. PD affects overall level of cognitive functioning and, to a lesser extent, DRS Initiation/Perseveration, Construction, and Attention. Diminished memory was primarily related to depression. Treatment of depression may ameliorate aspects of cognitive dysfunction in the PD patient with depression. PMID- 11884653 TI - Behavior in Huntington's disease: dissociating cognition-based and mood-based changes. AB - The authors examined the relationship of three dimensions of behavioral change (Apathy, Depression, and Irritability) measured by the Problem Behaviors Assessment for Huntington's Disease (PBA-HD) to cognitive and motor indices of disease severity. The Apathy subscale was highly correlated with both cognitive and motor impairment; the Irritability and Depression subscales were not. The findings suggest that certain behavioral alterations are intrinsic to the evolution and progression of HD, whereas others are more variable and are independent of other indices of disease progression. PMID- 11884654 TI - Frontal cortex atrophy predicts cognitive impairment in multiple sclerosis. AB - The association between regional measures of cortical atrophy and neuropsychological (NP) dysfunction was studied in 35 multiple sclerosis (MS) patients. Patients underwent neurological examination, MRI, and NP testing. Blind quantitative MRI analysis yielded total T(2) lesion area (TLA) and third ventricle width (3VW). Cortical atrophy, rated by blind visual inspection, was more extensive in superior frontal and parietal cortices than in other regions. No MRI measures were correlated with depression scores. TLA and 3VW were significantly correlated with each NP test. Cortical atrophy measures for bilateral superior frontal cortex were retained in regression models predicting impairments in verbal learning, spatial learning, attention, and conceptual reasoning. The authors conclude that cerebral atrophy predicts NP impairment while accounting for the influence of TLA or 3VW. Regions of cortex most susceptible to atrophic and cognitive changes in MS are the right and left superior frontal lobes. PMID- 11884655 TI - Addiction denial and cognitive dysfunction: a preliminary investigation. AB - This study explored the proposition that denial of addiction is often more a product of cognitive failure due to cerebral dysfunction than an emotion-driven rejection of the truth. Forty-four subjects were studied in an inpatient alcohol rehabilitation program. Denial was defined as the proportion of standardized denial-related treatment goals established at admission that remained unachieved at discharge. Cognitive deficiencies were identified through neuropsychological assessments. Persistent denial was significantly correlated with greater impairment of executive function, verbal memory, visual inference, and mental speed. PMID- 11884657 TI - Impaired recognition of facial expressions of emotion in Alzheimer's disease. AB - Recognizing facial emotions is an important aspect of interpersonal communication that may be impaired in Alzheimer's disease (AD). The authors examined facial emotion matching, facial emotion labeling, and same--different emotion differentiation in AD patients, healthy elderly volunteers, and elderly, nondemented psychiatric outpatients. Compared with both control groups, AD patients were significantly impaired on all three measures. AD patients were also impaired on a facial identity matching task. Using facial identity matching scores as a covariate provided evidence suggesting the facial emotion processing deficit may be independent of impairment in nonemotional face processing. AD patients also had selective impairment in labeling facial expressions of sadness. The authors conclude that patients with AD have deficits in recognizing facial emotions, which may be independent of their impairment in recognizing nonemotional features of faces. PMID- 11884656 TI - Learning and recall in subjects at genetic risk for Alzheimer's disease. AB - Deficits in delayed recall of learned information may be an early marker of Alzheimer's disease (AD). The apolipoprotein E E4 allele and a positive family history (FH) are both genetic risk factors for AD. The authors cross-sectionally compared performance on the California Verbal Learning Test (CVLT) in 153 prospectively recruited normal elderly subjects (mean age 67 years, mean MMSE=28) stratified by genetic risk into four groups (E4+/FH+, E4+/FH-, E4-/FH+, E4-/FH-). Neither FH nor E4 status affected performance, except on List B (a distraction word list), on which the FH+ group performed worse. The high-risk group (E4+/FH+) also performed worse on List B than the low-risk group (E4-/FH-) but did not differ on other measures. Memory impairments associated with genetic or family history risk may not manifest until the person is much closer to the onset age of AD. PMID- 11884658 TI - Impaired visuomotor function in schizophrenic patients compared with control subjects. AB - Visuomotor function was studied in 36 schizophrenic patients treated with atypical antipsychotics and in 22 control subjects. Patients showed significant disturbances in ability to control movement direction when tracing objects on screen and in keeping pace with a moving target in tracking tests. The impairments were not related to medication dose or to extrapyramidal side effects. Visuomotor impairment may be part of illness-related pathology in schizophrenia. PMID- 11884659 TI - Transcranial magnetic stimulation of left auditory cortex in patients with schizophrenia: effects on hallucinations and neurocognition. AB - The effects of transcranial magnetic stimulation (TMS) on hallucination severity and neurocognition were studied in 9 medication-resistant hallucinating patients. A statistically significant improvement was observed on a hallucination scale after 10 days of TMS at the left auditory cortex. PMID- 11884660 TI - Apolipoprotein E gender effects on cognitive performance in age-associated memory impairment. AB - Among 100 individuals with age-associated memory impairment (AAMI), APOE E4 carriers performed worse on memory. However, when subjects were considered by gender, this effect was only observed in females. APOE E4 may have a more robust cognitive influence on female than on male individuals with AAMI. PMID- 11884662 TI - Slowly progressive alexia. PMID- 11884661 TI - Valproate for catatonia: need for caution in patients on SSRIs and antipsychotics. PMID- 11884664 TI - Lamotrigine--clozapine combination in refractory schizophrenia: three cases. PMID- 11884665 TI - A case of neuroleptic malignant syndrome with quetiapine. PMID- 11884667 TI - Obsessive-compulsive disorder associated with a left orbitofrontal infarct. PMID- 11884666 TI - Risperidone and refusal to eat after traumatic brain injury. PMID- 11884668 TI - Cortisol and reduced interhemispheric coupling between the left prefrontal and the right parietal cortex. PMID- 11884669 TI - A third arm on the chest: implications for the cortical reorganization theory of phantom limbs. PMID- 11884674 TI - Tangerine dreams: cloning of carotenoid isomerase from Arabidopsis and tomato. PMID- 11884675 TI - An emerging model of auxin transport regulation. PMID- 11884676 TI - Arabidopsis SHY2/IAA3 inhibits auxin-regulated gene expression. AB - In Arabidopsis, SHY2 encodes IAA3, a member of the auxin-induced Aux/IAA family. Gain-of-function mutations in SHY2/IAA3 cause enlarged cotyledons, short hypocotyls, and altered auxin-regulated root development. Here we show that the gain-of-function mutation shy2-2 decreases both the induction and repression of auxin-regulated genes, suggesting that SHY2/IAA3 acts as a negative regulator in auxin signaling. shy2-2 affects auxin induction of many previously characterized primary response genes, implying that it might repress primary auxin responses. In addition, shy2-2 also affects expression of multiple auxin-nonresponsive genes. Light regulates expression of SHY2/IAA3, suggesting a possible link between light and auxin response pathways. PMID- 11884677 TI - Identification of the carotenoid isomerase provides insight into carotenoid biosynthesis, prolamellar body formation, and photomorphogenesis. AB - Carotenoids are essential photoprotective and antioxidant pigments synthesized by all photosynthetic organisms. Most carotenoid biosynthetic enzymes were thought to have evolved independently in bacteria and plants. For example, in bacteria, a single enzyme (CrtI) catalyzes the four desaturations leading from the colorless compound phytoene to the red compound lycopene, whereas plants require two desaturases (phytoene and zeta-carotene desaturases) that are unrelated to the bacterial enzyme. We have demonstrated that carotenoid desaturation in plants requires a third distinct enzyme activity, the carotenoid isomerase (CRTISO), which, unlike phytoene and zeta-carotene desaturases, apparently arose from a progenitor bacterial desaturase. The Arabidopsis CRTISO locus was identified by the partial inhibition of lutein synthesis in light-grown tissue and the accumulation of poly-cis-carotene precursors in dark-grown tissue of crtISO mutants. After positional cloning, enzymatic analysis of CRTISO expressed in Escherichia coli confirmed that the enzyme catalyzes the isomerization of poly cis-carotenoids to all-trans-carotenoids. Etioplasts of dark-grown crtISO mutants accumulate acyclic poly-cis-carotenoids in place of cyclic all-trans-xanthophylls and also lack prolamellar bodies (PLBs), the lattice of tubular membranes that defines an etioplast. This demonstrates a requirement for carotenoid biosynthesis to form the PLB. The absence of PLBs in crtISO mutants demonstrates a function for this unique structure and carotenoids in facilitating chloroplast development during the first critical days of seedling germination and photomorphogenesis. PMID- 11884678 TI - Cloning of tangerine from tomato reveals a carotenoid isomerase essential for the production of beta-carotene and xanthophylls in plants. AB - Carotenoid biosynthesis in plants has been described at the molecular level for most of the biochemical steps in the pathway. However, the cis-trans isomerization of carotenoids, which is known to occur in vivo, has remained a mystery since its discovery five decades ago. To elucidate the molecular mechanism of carotenoid isomerization, we have taken a genetic map-based approach to clone the tangerine locus from tomato. Fruit of tangerine are orange and accumulate prolycopene (7Z,9Z,7'Z,9'Z-tetra-cis-lycopene) instead of the all trans-lycopene, which normally is synthesized in the wild type. Our data indicate that the tangerine gene, designated CRTISO, encodes an authentic carotenoid isomerase that is required during carotenoid desaturation. CRTISO is a redox-type enzyme structurally related to the bacterial-type phytoene desaturase CRTI. Two alleles of tangerine have been investigated. In tangerine(mic), loss of function is attributable to a deletion mutation in CRTISO, and in tangerine(3183), expression of this gene is impaired. CRTISO from tomato is expressed in all green tissues but is upregulated during fruit ripening and in flowers. The function of carotene isomerase in plants presumably is to enable carotenoid biosynthesis to occur in the dark and in nonphotosynthetic tissues. PMID- 11884679 TI - Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling. AB - The phytohormone abscisic acid (ABA) plays an essential role in adaptive stress responses. The hormone regulates, among others, the expression of numerous stress responsive genes. From various promoter analyses, ABA-responsive elements (ABREs) have been determined and a number of ABRE binding factors have been isolated, although their in vivo roles are not known. Here we report that the ABRE binding factors ABF3 and ABF4 function in ABA signaling. The constitutive overexpression of ABF3 or ABF4 in Arabidopsis resulted in ABA hypersensitivity and other ABA associated phenotypes. In addition, the transgenic plants exhibited reduced transpiration and enhanced drought tolerance. At the molecular level, altered expression of ABA/stress-regulated genes was observed. Furthermore, the temporal and spatial expression patterns of ABF3 and ABF4 were consistent with their suggested roles. Thus, our results provide strong in vivo evidence that ABF3 and ABF4 mediate stress-responsive ABA signaling. PMID- 11884680 TI - Short defective interfering RNAs of tombusviruses are not targeted but trigger post-transcriptional gene silencing against their helper virus. AB - Post-transcriptional gene silencing (PTGS) is a sequence-specific degradation mechanism that operates in almost all eukaryotic cells. In plants, double stranded RNA triggers PTGS, generating 21- to 25-nucleotide guide RNAs responsible for specific degradation of cognate mRNA. The double stranded RNA intermediates of replicating plant viruses often induce PTGS, leading to symptom attenuation. Here we demonstrate the role of PTGS in defective interfering (DI) RNA-mediated symptom attenuation in plants infected with Cymbidium ringspot tombusvirus (CymRSV). Analysis of 21- to 25-nucleotide RNAs in Nicotiana benthamiana infected with CymRSV indicated that PTGS was not spread homogeneously along the viral genome. The 21- to 25-nucleotide RNAs derived mainly from plus stranded RNA and likely arose from local basepaired structures. In contrast to helper viral RNA, short DI RNAs were not accessible to helper virus-induced RNA degradation guided by the 21- to 25-nucleotide RNAs. Our results suggest a model in which PTGS plays an important role in the selective accumulation and symptom attenuation mediated by DI RNAs. Because PTGS operates in a wide variety of different organisms, this model is applicable to DI RNA generation and accumulation in both plant and animal cells. PMID- 11884681 TI - Cryptochrome light signals control development to suppress auxin sensitivity in the moss Physcomitrella patens. AB - The blue light receptors termed cryptochromes mediate photomorphological responses in seed plants. However, the mechanisms by which cryptochrome signals regulate plant development remain obscure. In this study, cryptochrome functions were analyzed using the moss Physcomitrella patens. This moss has recently become known as the only plant species in which gene replacement occurs at a high frequency by homologous recombination. Two cryptochrome genes were identified in Physcomitrella, and single and double disruptants of these genes were generated. Using these disruptants, it was revealed that cryptochrome signals regulate many steps in moss development, including induction of side branching on protonema and gametophore induction and development. In addition, the disruption of cryptochromes altered auxin responses, including the expression of auxin inducible genes. Cryptochrome disruptants were more sensitive to external auxin than wild type in a blue light-specific manner, suggesting that cryptochrome light signals repress auxin signals to control plant development. PMID- 11884682 TI - The abscisic acid-related SNARE homolog NtSyr1 contributes to secretion and growth: evidence from competition with its cytosolic domain. AB - Syntaxins and other SNARE proteins are crucial for intracellular vesicle trafficking, fusion, and secretion. Previously, we isolated the syntaxin-related protein NtSyr1 (NtSyp121) from tobacco in a screen for abscisic acid-related signaling elements, demonstrating its role in determining the abscisic acid sensitivity of K(+) and Cl(-) channels in stomatal guard cells. NtSyr1 is localized to the plasma membrane and is expressed normally throughout the plant, especially in root tissues, suggesting that it might contribute to cellular homeostasis as well as to signaling. To explore its functions in vivo further, we examined stably transformed lines of tobacco that expressed various constructs of NtSyr1, including the full-length protein and a truncated fragment, Sp2, corresponding to the cytosolic domain shown previously to be active in suppressing ion channel response to abscisic acid. Constitutively overexpressing NtSyr1 yielded uniformly high levels of protein (>10 times the wild-type levels) and was associated with a significant enhancement of root growth in seedlings but not with any obvious phenotype in mature, well-watered plants. Similar transformations with constructs encoding the Sp2 fragment of NtSyr1 showed altered leaf morphology but gave only low levels of Sp2 fragment, suggesting a strong selective pressure against plants expressing this protein. High expression of the Sp2 fragment was achieved in stable transformants under the control of a dexamethasone-inducible promoter. Sp2 expression was correlated positively with altered cellular and tissue morphology in leaves and roots and with a cessation of growth in seedlings. Overexpression of the full-length NtSyr1 protein rescued the wild-type phenotype, even in plants expressing high levels of the Sp2 fragment, supporting the idea that the Sp2 fragment interfered specifically with NtSyr1 function by competing with NtSyr1 for its binding partners. To explore NtSyr1 function in secretion, we used a green fluorescent protein (GFP)-based section assay. When a secreted GFP marker was coexpressed with Sp2 in tobacco leaves, GFP fluorescence was retained in cytosolic reticulate and punctate structures. In contrast, in plants coexpressing secreted GFP and NtSyr1 or secreted GFP alone, no GFP fluorescence accumulated within the cells. A new yellow fluorescent protein-based secretion marker was used to show that the punctate structures labeled in the presence of Sp2 colocalized with a Golgi marker. These structures were not labeled in the presence of a dominant Rab1 mutant that inhibited transport from the endoplasmic reticulum to the Golgi. We propose that NtSyr1 functions as an element in SNARE-mediated vesicle trafficking to the plasma membrane and is required for cellular growth and homeostasis. PMID- 11884683 TI - Independently regulated neocentromere activity of two classes of tandem repeat arrays. AB - Tandem repeat arrays often are found in interstitial (i.e., normally gene-rich) regions on chromosomes. In maize, genes on abnormal chromosome 10 induce the tandem repeats that make up knobs to move poleward on the meiotic spindle. This so-called neocentromere activity results in the preferential recovery, or meiotic drive, of the knobs in progeny. Here we show that two classes of repeats differ in their capacity to form neocentromeres and that their motility is controlled in trans by at least two repeat-specific activators. Microtubule dynamics appear to contribute little to the movement of neocentromeres (they are active in the presence of taxol), suggesting that the mechanism of motility involves microtubule-based motors. These data suggest that maize knob repeats and their binding proteins have coevolved to ensure their preferential recovery in progeny. Neocentromere-mediated drive provides a plausible mechanism for the evolution and maintenance of repeat arrays that occur in interstitial positions. PMID- 11884684 TI - AXR1-ECR1-dependent conjugation of RUB1 to the Arabidopsis Cullin AtCUL1 is required for auxin response. AB - Mutations in the AXR1 gene result in a reduction in auxin response and diverse defects in auxin-regulated growth and development. In a previous study, we showed that AXR1 forms a heterodimer with the ECR1 protein. This enzyme activates the ubiquitin-related protein RUB1 in vitro. Furthermore, we showed that the Skp1 Cul1/Cdc53-F-box (SCF) subunit AtCUL1 is modified by RUB1 in vivo. In this report, we demonstrate that the formation of RUB-AtCUL1 is dependent on AXR1 and ECR1 in vivo. The expression of AXR1 and ECR1 is restricted to zones of active cell division and cell elongation, consistent with their role in growth regulation. These results provide strong support for a model in which RUB conjugation of AtCUL1 affects the function of SCF E3s that are required for auxin response. PMID- 11884685 TI - Large-scale structure-function analysis of the Arabidopsis RPM1 disease resistance protein. AB - The Arabidopsis RPM1 gene confers resistance against Pseudomonas syringae expressing either the AvrRpm1 or the AvrB type III effector protein. We present an exhaustive genetic screen for mutants that no longer recognize avrRpm1. Using an inducible avrRpm1 expression system, we identified 110 independent mutations. These mutations represent six complementation groups. None discriminates between avrRpm1 and avrB recognition. We identified 95 rpm1 alleles and present a detailed structure--function analysis of the RPM1 protein. Several rpm1 mutants retain partial function, and we deduce that their residual activity is dependent on the level of avrRpm1 signal. In these mutants, the hypersensitive response remains activated if the signal goes above a certain threshold. Missense mutations in rpm1 are highly enriched in the nucleotide binding domain, suggesting that this region plays a key role either in the hypersensitive response associated with RPM1 activation or in RPM1 stability. Cluster analysis of rpm1 alleles defines functionally important residues that are highly conserved between nucleotide binding site leucine-rich repeat R proteins and those that are unique to RPM1. Regions of RPM1 to which no loss-of-function alleles map may represent domains in which variation is tolerated and may contribute to the evolution of new R gene specificities. PMID- 11884687 TI - The putative plasma membrane Na(+)/H(+) antiporter SOS1 controls long-distance Na(+) transport in plants. AB - The salt tolerance locus SOS1 from Arabidopsis has been shown to encode a putative plasma membrane Na(+)/H(+) antiporter. In this study, we examined the tissue-specific pattern of gene expression as well as the Na(+) transport activity and subcellular localization of SOS1. When expressed in a yeast mutant deficient in endogenous Na(+) transporters, SOS1 was able to reduce Na(+) accumulation and improve salt tolerance of the mutant cells. Confocal imaging of a SOS1-green fluorescent protein fusion protein in transgenic Arabidopsis plants indicated that SOS1 is localized in the plasma membrane. Analysis of SOS1 promoter-beta-glucuronidase transgenic Arabidopsis plants revealed preferential expression of SOS1 in epidermal cells at the root tip and in parenchyma cells at the xylem/symplast boundary of roots, stems, and leaves. Under mild salt stress (25 mM NaCl), sos1 mutant shoot accumulated less Na(+) than did the wild-type shoot. However, under severe salt stress (100 mM NaCl), sos1 mutant plants accumulated more Na(+) than did the wild type. There also was greater Na(+) content in the xylem sap of sos1 mutant plants exposed to 100 mM NaCl. These results suggest that SOS1 is critical for controlling long-distance Na(+) transport from root to shoot. We present a model in which SOS1 functions in retrieving Na(+) from the xylem stream under severe salt stress, whereas under mild salt stress it may function in loading Na(+) into the xylem. PMID- 11884686 TI - The protein encoded by oncogene 6b from Agrobacterium tumefaciens interacts with a nuclear protein of tobacco. AB - The 6b gene in the T-DNA from Agrobacterium has oncogenic activity in plant cells, inducing tumor formation, the phytohormone-independent division of cells, and alterations in leaf morphology. The product of the 6b gene appears to promote some aspects of the proliferation of plant cells, but the molecular mechanism of its action remains unknown. We report here that the 6b protein associates with a nuclear protein in tobacco that we have designated NtSIP1 (for Nicotiana tabacum 6b-interacting protein 1). NtSIP1 appears to be a transcription factor because its predicted amino acid sequence includes two regions that resemble a nuclear localization signal and a putative DNA binding motif, which is similar in terms of amino acid sequence to the triple helix motif of rice transcription factor GT 2. Expression in tobacco cells of a fusion protein composed of the DNA binding domain of the yeast GAL4 protein and the 6b protein activated the transcription of a reporter gene that was under the control of a chimeric promoter that included the GAL4 upstream activating sequence and the 35S minimal promoter of Cauliflower mosaic virus. Furthermore, nuclear localization of green fluorescent protein-fused 6b protein was enhanced by NtSIP1. A cluster of acidic residues in the 6b protein appeared to be essential for nuclear localization and for transactivation as well as for the hormone-independent growth of tobacco cells. Thus, it seems possible that the 6b protein might function in the proliferation of plant cells, at least in part, through an association with NtSIP1. PMID- 11884688 TI - Age-related resistance in Arabidopsis is a developmentally regulated defense response to Pseudomonas syringae. AB - Age-related resistance (ARR) has been observed in a number of plant species; however, little is known about the biochemical or molecular mechanisms involved in this response. Arabidopsis becomes more resistant, or less susceptible, to virulent Pseudomonas syringae (pv tomato or maculicola) as plants mature (in planta bacterial growth reduction of 10- to 100-fold). An ARR-like response also was observed in response to certain environmental conditions that accelerate Arabidopsis development. ARR occurs in the Arabidopsis mutants pad3-1, eds7-1, npr1-1, and etr1-4, suggesting that ARR is a distinct defense response, unlike the induced systemic resistance or systemic acquired resistance responses. However, three salicylic acid (SA) accumulation-deficient plant lines, NahG, sid1, and sid2, did not exhibit ARR. A heat-stable antibacterial activity was detected in intercellular washing fluids in response to Pst inoculation in wild type ARR-competent plants but not in NAHG: These data suggest that the ability to accumulate SA is necessary for the ARR response and that SA may act as a signal for the production of the ARR-associated antimicrobial compound(s) and/or it may possess direct antibacterial activity against P. syringae. PMID- 11884689 TI - The dominance of alleles controlling self-incompatibility in Brassica pollen is regulated at the RNA level. AB - Self-incompatibility (SI) in Brassica is controlled sporophytically by the multiallelic S-locus. The SI phenotype of pollen in an S-heterozygote is determined by the relationship between the two S-haplotypes it carries, and dominant/recessive relationships often are observed between the two S-haplotypes. The S-locus protein 11 (SP11, also known as the S-locus cysteine-rich protein) gene has been cloned from many pollen-dominant S-haplotypes (class I) and shown to encode the pollen S-determinant. However, SP11 from pollen-recessive S haplotypes (class II) has never been identified by homology-based cloning strategies, and how the dominant/recessive interactions between the two classes occur was not known. We report here the identification and molecular characterization of SP11s from six class II S-haplotypes of B. rapa and B. oleracea. Phylogenetic analysis revealed that the class II SP11s form a distinct group separated from class I SP11s. The promoter sequences and expression patterns of SP11s also were different between the two classes. The mRNA of class II SP11, which was detected predominantly in the anther tapetum in homozygotes, was not detected in the heterozygotes of class I and class II S-haplotypes, suggesting that the dominant/recessive relationships of pollen are regulated at the mRNA level of SP11s. PMID- 11884691 TI - AIDS in Southern Africa. PMID- 11884690 TI - Characterization of phenylpropene O-methyltransferases from sweet basil: facile change of substrate specificity and convergent evolution within a plant O methyltransferase family. AB - Some basil varieties are able to convert the phenylpropenes chavicol and eugenol to methylchavicol and methyleugenol, respectively. Chavicol O-methyltransferase (CVOMT) and eugenol O-methyltransferase (EOMT) cDNAs were isolated from the sweet basil variety EMX-1 using a biochemical genomics approach. These cDNAs encode proteins that are 90% identical to each other and very similar to several isoflavone O-methyltransferases such as IOMT, which catalyzes the 4'-O methylation of 2,7,4'-trihydroxyisoflavanone. On the other hand, CVOMT1 and EOMT1 are related only distantly to (iso)eugenol OMT from Clarkia breweri, indicating that the eugenol O-methylating enzymes in basil and C. breweri evolved independently. Transcripts for CVOMT1 and EOMT1 were highly expressed in the peltate glandular trichomes on the surface of the young basil leaves. The CVOMT1 and EOMT1 cDNAs were expressed in Escherichia coli, and active proteins were produced. CVOMT1 catalyzed the O-methylation of chavicol, and EOMT1 also catalyzed the O-methylation of chavicol with equal efficiency to that of CVOMT1, but it was much more efficient in O-methylating eugenol. Molecular modeling, based on the crystal structure of IOMT, suggested that a single amino acid difference was responsible for the difference in substrate discrimination between CVOMT1 and EOMT1. This prediction was confirmed by site-directed mutagenesis, in which the appropriate mutants of CVOMT1 (F260S) and EOMT1 (S261F) were produced that exhibited the opposite substrate preference relative to the respective native enzyme. PMID- 11884692 TI - Clinical application of findings of clinical trials and systematic reviews. PMID- 11884693 TI - Data and models determine treatment proposals--an illustration from meta analysis. AB - A relevant problem in meta-analysis concerns the possible heterogeneity between trial results. If a test of heterogeneity is not significant the trials are often considered to be "homogeneous" and the individual trial results are replaced by an overall mean effect size and its confidence interval ("equal effects model"). If the trials are heterogeneous the individual trial effect sizes are conserved ("fixed effects model"). In a more flexible approach ("random effects model"), each trial makes use of knowledge from the other trials so individual effect sizes are "shrunken" towards an overall mean effect size. The more flexible tool may be useful for doctors involved in a trial when the outcome of their individual trial differs markedly from the overall mean effect size. Where a particular trial result is opposite in direction to the overall mean result, a conflict may arise: should a new patient be treated with the new method or not? The more flexible position and a graphical comparison of the three approaches are likely to be helpful in guiding the decision. Applying different models to the same data may lead to apparently paradoxical results: an individual trial result may be interpreted to be beneficial or harmful depending on the choice of model. PMID- 11884694 TI - Liver transplantation for chronic liver disease: advances and controversies in an era of organ shortages. AB - Since liver transplantation was first performed in 1968 by Starzl et al, advances in case selection, liver surgery, anaesthetics, and immunotherapy have significantly increased the indications for and success of this operation. Liver transplantation is now a standard therapy for many end stage liver disorders as well as acute liver failure. However, while demand for cadaveric organ grafts has increased, in recent years the supply of organs has fallen. This review addresses current controversies resulting from this mismatch. In particular, methods for increasing graft availability and difficulties arising from transplantation in the context of alcohol related cirrhosis, primary liver tumours, and hepatitis C are reviewed. Together these three indications accounted for 42% of liver transplants performed for chronic liver disease in the UK in 2000. Ethical frameworks for making decisions on patients' suitability for liver transplantation have been developed in both the USA and the UK and these are also reviewed. PMID- 11884696 TI - Primary care referral protocol for carpal tunnel syndrome. AB - Carpal tunnel syndrome is an extremely common upper limb nerve compression syndrome, widely distributed in the community. There are a variety of treatment options which may be applied to the syndrome, depending on the severity of symptoms. Some options are available in a primary care setting, others require secondary referral. This paper is a detailed review of the available literature and provides a protocol that could be used to assist in the referral of patients from primary care. PMID- 11884695 TI - Understanding the pathology of schizophrenia: recent advances from the study of the molecular architecture of postmortem CNS tissue. AB - The use of central nervous system (CNS) tissue obtained postmortem has long underpinned efforts to understand the neurobiology of schizophrenia, but the ability to use such tissue in conjunction with a wide variety of methodologies has seen a renaissance of interest in this area of research. Recent findings have shown changes in markers in a number of neurotransmitter systems in the brains of subjects with schizophrenia which include the dopaminergic, serotonergic, cholinergic, glutamatergic, and GABAergic systems of the CNS. Many of these changes also appear to be regionally specific, and abnormalities in non neurotransmitter specific pathways have been found in schizophrenia. Changes in the neurotransmitter release pathways in schizophrenia may be important in the pathology of the illness, and recent findings suggest that abnormalities in the Wnt pathway, which controls transcription selectivity in cells, may be involved. Studies using CNS material obtained postmortem clearly show that the pathology of schizophrenia is complex while the polygenetic nature of the illness may be adding to this complexity. PMID- 11884697 TI - Living kidney donation: a comparison of laparoscopic and conventional open operations. AB - Laparoscopic donor nephrectomy has the potential to lessen the burden placed on live kidney donors. This study describes the first British comparison of donor morbidity and recovery following conventional open donor nephrectomy (ODN) and laparoscopic donor nephrectomy (LDN). An initial series of LDN (n=20) was compared to a historical control group of ODN (n=34). Laparoscopic operations were performed via a transperitoneal approach, the kidney being removed through a 6--12 cm Pfannensteil incision. Open operations were performed using a retroperitoneal flank approach with resection of the 12th rib. Postoperatively, donors were managed with a patient controlled analgesia system. LDN was associated with shorter mean (SD) inpatient stay (6 (2) v 4 (1) days; p=0.0001) and lower parenteral narcotic requirements (morphine 179 (108) v 67 (54) mg; p=0.0001). Laparoscopic donors started driving their cars sooner (2 (1.5) v 6 (4) weeks; p=0.0001) and returned to work more quickly (5 (3) v 12 (6) weeks; p=0.0001) than open nephrectomy donors. There were no differences in recipient serum creatinine levels at three months post-transplant but two recipients of transplant kidneys retrieved laparoscopically (10%) developed ureteric obstruction, whereas this complication did not occur after ODN (p=0.13). LDN is associated with less postoperative pain and a substantial improvement in donor recovery times. It is not yet clear whether or not the outcome of the recipient kidney transplants are the same after ODN and LDN and much more experience is required before the place of this new technique can be defined. PMID- 11884698 TI - Misinterpretation of the chest x ray as a factor in the delayed diagnosis of lung cancer. AB - All patients in 1997 with a histologically proved diagnosis of lung cancer in Castle Hill Hospital in whom a full set of case notes and x rays could be retrieved were studied. All previous chest x rays were reviewed by a consultant chest physician and a radiologist, who were blinded to the eventual site of the lesion and the point at which a suspicious abnormality first appeared. Case notes were inspected to clarify the cause of any error. Fifty eight patients were eligible, 28 of whom had previous chest x rays. Of these 14 were found to be abnormal. A significant difference (p=0.007) in time from diagnosis to death was found between those with a missed abnormality, median (interquartile range, IQR) 105 (55-219) days and those with no previous abnormality, median (IQR) 260 (137 512) days. In the 14 in whom the diagnosis was missed the median (IQR) delay from first abnormal chest x ray to the eventual diagnostic x ray was 101 (48-339) days. A significant difference (p=0.001) was also found between the median (IQR) time from first abnormal chest x ray to start of treatment between those with missed abnormalities, 155 (115-376) days, and those with no previous abnormality on chest x ray, 51 (44-77) days. The most common reason (47%) for the diagnosis to be missed was failure of the radiologist reporting the x ray to recognise the abnormality. It is not unusual to find previous significant radiological abnormalities in patients in whom a diagnosis of lung cancer is later made. This leads to a diagnostic delay which has a significant effect on time to initiation of treatment and palliation of symptoms, although not necessarily to eventual outcome. PMID- 11884699 TI - Audit of a nurse endoscopist based one stop dyspepsia clinic. AB - As a response to the UK Health Department's "two week cancer wait" initiative a one stop dyspepsia clinic based on a nurse endoscopist was introduced, and the first 100 cases attending this clinic have been audited. After referral on a purpose designed form, patients were assessed by a gastroenterologist and then investigated at the same visit--where possible and appropriate--by endoscopy or ultrasound scan. All endoscopies were performed by a trained nurse specialist. Of the 100 patients, 84 were gastroscoped the same day and 11 had an ultrasound scan. Inappropriate tests were avoided in 16% of referrals. The commonest endoscopic diagnoses were minor oesophageal or gastroduodenal inflammation (64% of gastroscopies). Only six oesophageal or gastric cancers were found--all at an advanced stage--and three further malignancies were diagnosed. Only a minority (12%) of the patients with "alarm symptoms" had cancer. The waiting time for an appointment rose progressively during the first six months of the clinic. The system was popular with patients as most of them (70%) were dealt with at a single hospital attendance. Basing the endoscopy practice on a trained nurse specialist not only facilitated the creation of the service by maximising the use of scarce resources, but also improved communication and overall management of patients. PMID- 11884700 TI - Parasites in fine needle breast aspirates--assessment of host tissue response. AB - Parasites in the human breast are uncommon but not rare. Cysticercus and filariasis in fine needle breast aspirates have been documented and their cytomorphology is well characterised. However, the host tissue response to these parasites and the factors responsible for their initiation are not clear. Over a 21 year period, 28 cases of breast parasites (16 cases of cysticercus and 12 of filariasis), diagnosed by fine needle aspiration cytology, were reviewed to assess the host tissue response. PMID- 11884701 TI - Aortic root abscess presenting as unstable angina due to extrinsic compression of the left coronary artery. AB - Coronary ischaemia in acute endocarditis is usually due to pre-existing coronary disease or occasionally as a result of embolism from vegetations. A 68 year old man with known mixed aortic valve disease presented with a four week history of progressive exertional angina, which became unstable. He was apyrexial with no peripheral signs of endocarditis. Three sets of blood cultures were negative. Transthoracic echocardiography with suboptimal windows confirmed moderate mixed aortic valve disease. Marked reversible ST segment depression with angina recurred at rest. Aortography showed severe aortic regurgitation with a distorted aortic root. Coronary angiography showed severe proximal narrowing of the left anterior descending and circumflex arteries with an unusual long and tapering contour. Emergency surgery revealed a large anterior aortic root abscess which had destroyed the left and right coronary cusps. Aortic root abscess and other rare causes of extrinsic coronary compression are discussed. PMID- 11884702 TI - Pseudomyxoma peritonei. AB - Pseudomyxoma peritonei is a relatively rare and poorly understood condition in which mucus accumulates within the peritoneal cavity. The presence of cells in the mucin, either inflammatory or neoplastic, distinguishes it from simple acellular mucus ascites caused by mucinous spillage. There is widespread seeding of the peritoneal and omental surfaces with a heavy cancerous glaze. This is principally a complication of borderline or malignant neoplasm of the ovary and/or appendix. This paper describes two cases of previously healthy women who both presented with an acute abdomen, and were diagnosed postoperatively with pseudomyxoma peritonei. In addition, literature on the clinical presentation, diagnostic procedures, and treatment options has been briefly reviewed. PMID- 11884703 TI - Isolated splenic metastasis from carcinoma of the breast. AB - A case of isolated splenic metastasis from carcinoma of the breast in a 54 year old woman, two years after treatment for breast carcinoma, is presented. There was no involvement of other organs like liver, bone, lungs, etc. The patient underwent splenectomy and recovered without any complications. This case is being reported because of the rarity of the lesion. PMID- 11884704 TI - Sister Joseph's nodule. PMID- 11884706 TI - Human Rights Act 1998 and mental health legislation: implications for the management of mentally ill patients. AB - In the management of mentally ill patients, there is a tension between protecting the rights of individual patients and safeguarding public safety. The Human Rights Act 1998 emphasises on the former while two recent white papers focus on the latter. This article first examines the extent to which the Mental Health Act 1983 is consistent with the Human Rights Act. It argues that while the recent white papers exploit the gaps in the judgments given by the European courts, its compatibility with human rights is very doubtful. The practical implications of the Human Rights Act for doctors are discussed. PMID- 11884705 TI - Integrated care pathways for vascular surgery: an analysis of the first 18 months. AB - OBJECTIVES: A review of the use of previously described integrated care pathways (ICPs) established for three elective vascular surgical procedures. DESIGN: A retrospective analysis of information gathered prospectively over an initial 18 month period of use of vascular surgical ICPs. SUBJECT: Patients admitted to a single vascular unit for "open" repair of abdominal aortic aneurysm (AAA), carotid endarterectomy, or femoropopliteal bypass grafting. METHODS: An analysis of variance data, length of stay, and costings after the use of ICPs, compared with previous clinical practice. RESULTS: Variance data were gathered for each of the three procedures. Variances of medication prescribing and delays in discharge were common to all procedures. In particular: (i) gastrointestinal complications were more specific to AAA repair and (ii) wound drains were removed a day later than originally proposed after femoropopliteal bypass. Overall, improved efficiency due to use of ICPs reduced the length of stay for all procedures, which was reflected in a potential cost saving of some 25%. CONCLUSION: There are clear benefits to the use of ICPs, resulting in more structured, efficient, and cost effective patient care. Recommended changes to current practice based on variance analysis will require continued audit to sustain this "evidence based" approach. PMID- 11884707 TI - An uncommon cause of lumbar radiculopathy. PMID- 11884708 TI - A woman with painless burns. PMID- 11884709 TI - Massive haemoptysis in a young woman. PMID- 11884710 TI - A painful and deformed wrist. PMID- 11884711 TI - An unusual presentation of calvarial tuberculosis. PMID- 11884717 TI - Redox regulation of forkhead proteins through a p66shc-dependent signaling pathway. AB - Genetic determinants of longevity include the forkhead-related transcription factor DAF-16 in the worm Caenorhabditis elegans and the p66shc locus in mice. We demonstrate that p66shc regulates intracellular oxidant levels in mammalian cells and that hydrogen peroxide can negatively regulate forkhead activity. In p66shc-/ cells, the activity of the mammalian forkhead homolog FKHRL1 is increased and redox-dependent forkhead inactivation is reduced. In addition, expression of FKHRL1 results in an increase in both hydrogen peroxide scavenging and oxidative stress resistance. These results demonstrate an important functional relation between three distinct elements linked to aging: forkhead proteins, p66shc, and intracellular oxidants. PMID- 11884718 TI - Crystal structure of the extracellular segment of integrin alpha Vbeta3 in complex with an Arg-Gly-Asp ligand. AB - The structural basis for the divalent cation-dependent binding of heterodimeric alphabeta integrins to their ligands, which contain the prototypical Arg-Gly-Asp sequence, is unknown. Interaction with ligands triggers tertiary and quaternary structural rearrangements in integrins that are needed for cell signaling. Here we report the crystal structure of the extracellular segment of integrin alphaVbeta3 in complex with a cyclic peptide presenting the Arg-Gly-Asp sequence. The ligand binds at the major interface between the alphaV and beta3 subunits and makes extensive contacts with both. Both tertiary and quaternary changes are observed in the presence of ligand. The tertiary rearrangements take place in betaA, the ligand-binding domain of beta3; in the complex, betaA acquires two cations, one of which contacts the ligand Asp directly and the other stabilizes the ligand-binding surface. Ligand binding induces small changes in the orientation of alphaV relative to beta3. PMID- 11884719 TI - The effect of algal symbionts on the accuracy of Sr/Ca paleotemperatures from coral. AB - The strontium-to-calcium ratio (Sr/Ca) of reef coral skeleton is commonly used as a paleothermometer to estimate sea surface temperatures (SSTs) at crucial times in Earth's climate history. However, these estimates are disputed, because uptake of Sr into coral skeleton is thought to be affected by algal symbionts (zooxanthellae) living in the host tissue. Here, we show that significant distortion of the Sr/Ca temperature record in coral skeleton occurs in the presence of algal symbionts. Seasonally resolved Sr/Ca in coral without symbionts reflects local SSTs with a temperature sensitivity equivalent to that of laboratory aragonite precipitated at equilibrium and the nighttime skeletal deposits of symbiotic reef corals. However, up to 65% of the Sr/Ca variability in symbiotic skeleton is related to symbiont activity and does not reflect water temperature. PMID- 11884720 TI - To publish or not to publish. PMID- 11884721 TI - Nuclear fusion. 'Bubble fusion' paper generates a tempest in a beaker. PMID- 11884722 TI - Infectious disease. New culprit emerges in river blindness. PMID- 11884723 TI - AIDS research. Delays jeopardize Italian program. PMID- 11884724 TI - Paleontology. Earliest signs of life just oddly shaped crud? PMID- 11884725 TI - Astronomy. Stellar flares illuminate young sun's outbursts. PMID- 11884726 TI - Neutrino detection. Japan hopes casings will do the trick. PMID- 11884727 TI - Stem cell research. Canada gives OK for new cell lines. PMID- 11884728 TI - Animal behavior. Guppy sex and gluttony guided by orange glow. PMID- 11884729 TI - Denmark. Greens see red over revisionist's new job. PMID- 11884731 TI - Stem cells. In the Mideast, pushing back the stem cell frontier. PMID- 11884730 TI - Genomics. Taking aim at Celera's shotgun. PMID- 11884732 TI - Stem cells. Are any two cell lines the same? PMID- 11884733 TI - 2002 Ocean Sciences Meeting. Salt fingers mix the sea. PMID- 11884734 TI - 2002 Ocean Sciences Meeting. Mussels on the move. PMID- 11884735 TI - 2002 Ocean Sciences Meeting. Coastal cool-down. PMID- 11884736 TI - New York Academy of Sciences. How fast can an old dog learn new tricks? PMID- 11884737 TI - Public health. Decision-making when science is ambiguous. PMID- 11884740 TI - Radical chemistry. From reactive intermediates to stable compounds. PMID- 11884738 TI - Structural biology. PMF through the redox loop. PMID- 11884739 TI - Neurobiology. What the synapse tells the neuron. PMID- 11884741 TI - Transcription. Unlocking the gates to gene expression. PMID- 11884742 TI - Solid state chemistry. A mixed oxide-hydride perovskite. PMID- 11884743 TI - Nuclear fusion. Evidence for nuclear reactions in imploding bubbles. PMID- 11884744 TI - Granular materials. Mixing and de-mixing. PMID- 11884745 TI - Molecular chaperones in the cytosol: from nascent chain to folded protein. AB - Efficient folding of many newly synthesized proteins depends on assistance from molecular chaperones, which serve to prevent protein misfolding and aggregation in the crowded environment of the cell. Nascent chain--binding chaperones, including trigger factor, Hsp70, and prefoldin, stabilize elongating chains on ribosomes in a nonaggregated state. Folding in the cytosol is achieved either on controlled chain release from these factors or after transfer of newly synthesized proteins to downstream chaperones, such as the chaperonins. These are large, cylindrical complexes that provide a central compartment for a single protein chain to fold unimpaired by aggregation. Understanding how the thousands of different proteins synthesized in a cell use this chaperone machinery has profound implications for biotechnology and medicine. PMID- 11884746 TI - Prevention of inhalational anthrax in the U.S. outbreak. PMID- 11884747 TI - Molecular basis of proton motive force generation: structure of formate dehydrogenase-N. AB - The structure of the membrane protein formate dehydrogenase-N (Fdn-N), a major component of Escherichia coli nitrate respiration, has been determined at 1.6 angstroms. The structure demonstrates 11 redox centers, including molybdopterin guanine dinucleotides, five [4Fe-4S] clusters, two heme b groups, and a menaquinone analog. These redox centers are aligned in a single chain, which extends almost 90 angstroms through the enzyme. The menaquinone reduction site associated with a possible proton pathway was also characterized. This structure provides critical insights into the proton motive force generation by redox loop, a common mechanism among a wide range of respiratory enzymes. PMID- 11884748 TI - Evidence for nuclear emissions during acoustic cavitation. AB - In cavitation experiments with deuterated acetone, tritium decay activity above background levels was detected. In addition, evidence for neutron emission near 2.5 million electron volts was also observed, as would be expected for deuterium deuterium fusion. Control experiments with normal acetone did not result in tritium activity or neutron emissions. Hydrodynamic shock code simulations supported the observed data and indicated highly compressed, hot (10(6) to 10(7) kelvin) bubble implosion conditions, as required for nuclear fusion reactions. PMID- 11884749 TI - Spontaneous air-driven separation in vertically vibrated fine granular mixtures. AB - We report the observation of the spontaneous separation of vertically vibrated mixtures of fine bronze and glass spheres of similar diameters. At low frequencies and at sufficient vibrational amplitudes, a sharp boundary forms between a lower region of glass and an upper region of the heavier bronze. The boundary undergoes various oscillations, including periodic tilting motion, but remains extremely sharp. At higher frequencies, the bronze separates as a mid height layer between upper and lower glass regions, and the oscillations are largely absent. The mechanism responsible for the separation can be traced to the effect of air on the granular motion. PMID- 11884750 TI - Singlet diradicals: from transition states to crystalline compounds. AB - Singlet diradicals are usually not energy minima. As observed by femtosecond spectroscopy, they readily couple to form final sigma bonds. Substituent effects allow lifetimes to increase into the microsecond range. Taking advantage of the properties of hetero-elements, a diradical has been prepared that is indefinitely stable at room temperature. The availability of diradicals that can be handled under standard laboratory conditions will lead to further insight into their chemical and physical properties, raising the likelihood of practical applications, especially in the field of molecular materials such as electrical conductors and ferromagnets. PMID- 11884751 TI - The hydride anion in an extended transition metal oxide array: LaSrCoO3H0.7. AB - We present the synthesis and structural characterization of a transition metal oxide hydride, LaSrCoO3H0.7, which adopts an unprecedented structure in which oxide chains are bridged by hydride anions to form a two-dimensional extended network. The metal centers are strongly coupled by their bonding with both oxide and hydride ligands to produce magnetic ordering at temperatures up to at least 350 kelvin. The synthetic route is sufficiently general to allow the prediction of a new class of transition metal--containing electronic and magnetic materials. PMID- 11884752 TI - Water in Earth's lower mantle. AB - Secondary ion mass spectrometry measurements show that Earth's representative lower mantle minerals synthesized in a natural peridotitic composition can dissolve considerable amounts of hydrogen. Both MgSiO3-rich perovskite and magnesiowustite contain about 0.2 weight percent (wt%) H2O, and CaSiO3-rich perovskite contains about 0.4 wt% H2O. The OH absorption bands in Mg-perovskite and magnesiowustite were also confirmed with the use of infrared microspectroscopic measurements. Earth's lower mantle may store about five times more H2O than the oceans. PMID- 11884753 TI - Repeated and sudden reversals of the dipole field generated by a spherical dynamo action. AB - Using long-duration, three-dimensional magnetohydrodynamic simulation, we found that the magnetic dipole field generated by a dynamo action in a rotating spherical shell repeatedly reverses its polarity at irregular intervals (that is, punctuated reversal). Although the total convection energy and magnetic energy alternate between a high-energy state and a low-energy state, the dipole polarity can reverse only at high-energy states where the north-south symmetry of the convection pattern is broken and the columnar vortex structure becomes vulnerable. Another attractive finding is that the quadrupole mode grows, exceeding the dipole mode before the reversal; this may help to explain how Earth's magnetic field reverses. PMID- 11884754 TI - Antarctic krill under sea ice: elevated abundance in a narrow band just south of ice edge. AB - We surveyed Antarctic krill (Euphausia superba) under sea ice using the autonomous underwater vehicle Autosub-2. Krill were concentrated within a band under ice between 1 and 13 kilometers south of the ice edge. Within this band, krill densities were fivefold greater than that of open water. The under-ice environment has long been considered an important habitat for krill, but sampling difficulties have previously prevented direct observations under ice over the scale necessary for robust krill density estimation. Autosub-2 enabled us to make continuous high-resolution measurements of krill density under ice reaching 27 kilometers beyond the ice edge. PMID- 11884755 TI - The role of endosymbiotic Wolbachia bacteria in the pathogenesis of river blindness. AB - Parasitic filarial nematodes infect more than 200 million individuals worldwide, causing debilitating inflammatory diseases such as river blindness and lymphatic filariasis. Using a murine model for river blindness in which soluble extracts of filarial nematodes were injected into the corneal stroma, we demonstrated that the predominant inflammatory response in the cornea was due to species of endosymbiotic Wolbachia bacteria. In addition, the inflammatory response induced by these bacteria was dependent on expression of functional Toll-like receptor 4 (TLR4) on host cells. PMID- 11884756 TI - Mannose receptor-mediated regulation of serum glycoprotein homeostasis. AB - Carbohydrates are thought to function as tags that mark circulatory glycoproteins for rapid clearance. To examine the role of the mannose receptor (MR) in glycoprotein clearance, we generated mice genetically deficient in MR. MR-/- mice were defective in clearing proteins bearing accessible mannose and N acetylglucosamine residues and had elevated levels of eight different lysosomal hydrolases. Proteomic analysis of MR-/- and control mouse sera showed that an additional 4 out of 52 proteins identified were elevated in MR-/- serum. Each of these is up-regulated during inflammation and wound healing. Thus, MR appears to operate as an essential regulator of serum glycoprotein homeostasis. PMID- 11884757 TI - Coordination of PIC assembly and chromatin remodeling during differentiation induced gene activation. AB - We analyzed the ordered recruitment of factors to the human alpha1 antitrypsin promoter around the initial activation of the gene during enterocyte differentiation. We found that a complete preinitiation complex, including phosphorylated RNA pol II, was assembled at the promoter long before transcriptional activation. The histone acetyltransferases CBP and P/CAF were recruited subsequently, but local histone hyperacetylation was delayed. After transient recruitment of the human Brahma homolog hBrm, remodeling of the neighboring nucleosome coincided with transcription initiation. The results suggest that, at this promoter, chromatin reconfiguration is a defining step of the initiation process, acting after the assembly of the Pol II machinery. PMID- 11884759 TI - Dependence of EPSP efficacy on synapse location in neocortical pyramidal neurons. AB - Neurons receive thousands of synaptic inputs throughout elaborate dendritic trees. Here we determine the somatic impact of excitatory postsynaptic potentials (EPSPs) generated at known dendritic sites in neocortical pyramidal neurons. As inputs became more distal, somatic EPSP amplitude decreased, whereas use dependent depression increased. Despite marked attenuation (>40-fold), when coactivated within a narrow time window (approximately 10 milliseconds), distal EPSPs could directly influence action potential output following dendritic spike generation. These findings reveal that distal EPSPs are ineffective sources of background somatic excitation, but through coincidence detection have a powerful transient signaling role. PMID- 11884758 TI - Purkinje cell degeneration (pcd) phenotypes caused by mutations in the axotomy induced gene, Nna1. AB - The classical recessive mouse mutant, Purkinje cell degeneration (pcd), exhibits adult-onset degeneration of cerebellar Purkinje neurons, retinal photoreceptors, olfactory bulb mitral neurons, and selected thalamic neurons, and has defective spermatogenesis. Here we identify Nna1 as the gene mutated in the original pcd and two additional pcd alleles (pcd2J and pcd3J). Nna1 encodes a putative nuclear protein containing a zinc carboxypeptidase domain initially identified by its induction in spinal motor neurons during axonal regeneration. The present study suggests an unexpected molecular link between neuronal degeneration and regeneration, and its results have potential implications for neurodegenerative diseases and male infertility. PMID- 11884761 TI - Glucose tolerance, insulin secretion and insulin sensitivity in polycystic ovary syndrome. PMID- 11884760 TI - Parallel single-cell monitoring of receptor-triggered membrane translocation of a calcium-sensing protein module. AB - Time courses of translocation of fluorescently conjugated proteins to the plasma membrane were simultaneously measured in thousands of individual rat basophilic leukemia cells. We found that the C2 domain---a calcium-sensing, lipid-binding protein module that is an essential regulator of protein kinase C and numerous other proteins---targeted proteins to the plasma membrane transiently if calcium was released from internal stores, and persistently in response to entry of extracellular calcium across the plasma membrane. The C2 domain translocation time courses of stimulated cells clustered into only two primary modes. Hence, the reversible recruitment of families of signaling proteins from one cellular compartment to another is a rapid bifurcation mechanism for inducing discrete states of cellular signaling networks. PMID- 11884762 TI - Analysis and optimization of nutritional set-up for murine pancreatic acinar cells. AB - CONTEXT: Pancreatic acinar cell cultivation poses a serious problem due to limitations in the in vitro survival time despite variations of dissociation protocols, culture media and nutrient supplements. OBJECTIVE: To establish a long term culture of murine pancreatic acinar cells which retain their viability, monolayer formation and responsiveness to secretagogues. In order to investigate the mechanism of the short-life of acinar cells studied in vitro, we studied their survival under the influence of different supplements on nutrient media. INTERVENTIONS: Dissociated pancreatic acini were prepared from BALB/c mice pancreata by collagenase digestion supplemented with bovine serum albumin fraction V and soybean trypsin inhibitor. A nutrient set-up was designed for their long term survival in vitro. RESULTS: It was observed that mouse pancreatic acinar cells dissociated in presence of bovine serum albumin fraction V and soybean trypsin inhibitor result in 95% viability. Further cultivation of these acinar cells in Waymouth's MB 752/1 medium supplemented with 10% fetal calf serum (v/v), soybean trypsin inhibitor, bovine serum albumin, dexamethasone, and epidermal growth factor results in their survival for more than 6 days in culture with 85% viability, retention of the secretagogue responsiveness and formation of a monolayer without any extracellular matrix coating. CONCLUSIONS: Our study clearly demonstrates that the addition of soybean trypsin inhibitor to culture medium reduces zymogen granule fragility and acinar cell death, thus increasing their viability for sufficiently long periods. The present study offers an excellent, in vitro model for the investigation of exocrine dysfunction in response to acinar cell injury. PMID- 11884763 TI - A new classification plot for the C-peptide suppression test. AB - CONTEXT AND OBJECTIVE: To evaluate the C-peptide suppression test as a screening test in patients with symptoms of hypoglycemia as compared to the standard fasting test. DESIGN: Retrospective discriminant analysis of data from C-peptide suppression tests. SETTING: Clinical study. PATIENTS: Patients with insulinomas and patients without insulinomas but having symptoms compatible with hypoglycemia. INTERVENTIONS: The results from C-peptide suppression tests of 26 patients with insulinomas and 100 patients without insulinomas were compared. MAIN OUTCOME MEASURES: A classification plot which introduces two discriminant parameters for the C-peptide suppression test: the ratio of [blood glucose]/[C peptide] at the lowest C-peptide concentration and mean glycemia during insulin infusion. RESULTS: In patients with insulinomas, minimal serum C-peptide levels were higher (1.81+/- 0.87 ng/mL; median 1.83 ng/mL; maximal suppression 37 +/- 24% of basal C-peptide levels) as compared to patients without insulinoma (0.40 +/- 0.15 ng/mL; median 0.30 ng/mL; maximal suppression of 75 +/- 9%; P<0.001). Mean glycemia during the test was lower in patients with insulinomas (30.8 +/- 3.3 vs. 47.5 +/- 8.3 mg/dL; P<0.001) as was the [blood glucose]/[C-peptide] ratio (21.9 +/- 14.6 vs. 139.2 +/- 43.8; P<0.001). Discriminant analysis revealed a specificity of 96% to rule out the diagnosis of 'insulinoma' at a 1% probability threshold with a sensitivity of 100%. CONCLUSIONS: We developed a new classification plot for the C-peptide suppression test in order to accurately identify those patients whose symptoms of hypoglycemia are not due to endogenous hyperinsulinemia/insulinomas. Thus, the need for fasting tests and hospitalization costs can be reduced. PMID- 11884764 TI - Neurology and neuropathology of the pancreatic innervation. PMID- 11884765 TI - Trypsin-based laboratory methods and carboxypeptidase activation peptide in acute pancreatitis. AB - Acute pancreatitis is a common disease varying widely in severity. At present, there is no "gold standard" for the diagnosis of acute pancreatitis. Currently, the diagnosis of acute pancreatitis is based on measurements of serum amylase and/or lipase activity, which are considered unsatisfactory due to their low level of accuracy. Early identification of acute pancreatitis and especially detection of patients with a severe form of the disease is of utmost importance. Premature intrapancreatic activation of trypsinogen is a crucial early event in the pathophysiology of acute pancreatitis. The conversion of trypsinogen to active trypsin is mediated by the release of its activation peptide (TAP). The active trypsin is then able to activate other pancreatic zymogens (i.e. procarboxypeptidase) leading to tissue damage and eventually to autodigestion of the pancreas. To improve the laboratory diagnostics of AP, new methods have been developed to measure this primary pancreatic proteolytic insult. Here we review the current knowledge and clinical implications of trypsin based laboratory methods and carboxypeptidase activation peptide (CAPAP) in the diagnosis and severity assessment of acute pancreatitis. PMID- 11884766 TI - Increased early rejection rate after conversion from tacrolimus in kidney and pancreas transplantation. AB - CONTEXT: A successful immunosuppression regimen for combined kidney and pancreas transplants is tacrolimus, mycophenolate mofetil, and prednisone. However, not all patients tolerate these immunosuppressants especially tacrolimus. OBJECTIVE: To evaluate the efficacy of cyclosporine as a rescue agent for tacrolimus toxicity in combined kidney and pancreas transplants. DESIGN: Retrospective. SETTING: Single center. PATIENTS: Thirty-five combined kidney and pancreas transplants were performed between July 1994 and January 1999. All patients were insulin dependent diabetics with end-stage renal disease. Twenty-eight (mean age: 36 years and 57% female) were available with at least 12 month follow-up. INTERVENTIONS: Conversion to cyclosporine following renal (biopsy proven) or pancreatic dysfunction. MAIN OUTCOME MEASURES: Toxicity, rejection rate, and patient/transplant organ survival. RESULTS: Nineteen transplant recipients (68%) were continuously maintained on tacrolimus while nine (32%) required conversion to cyclosporine 75 +/- 20 days post-transplant. Reasons for conversion included: hyperglycemia (n=2), hemolytic-uremic syndrome (n=1), and severe tacrolimus nephrotoxicity (n=6). By 12 months post-transplant, the 19 patients maintained on tacrolimus had 5 rejections (26%). Three of the 9 patients (33%) converted to cyclosporine had an acute rejection prior to conversion. Seven of these 9 patients (78%; P=0.017 vs. patients maintained on tacrolimus) had rejections an average of 25 +/- 4 days post-conversion. Four of the 7 patients had no previous rejections prior to conversion. In spite of increased rejections, the 1- and 2 year patient/graft survivals were unchanged by converting. CONCLUSIONS: Converting to cyclosporine from tacrolimus was associated with an increased risk of acute rejection especially within the first 30 days post conversion. PMID- 11884767 TI - Pancreatic stone protein of pancreatic calculi in chronic calcified pancreatitis in man. AB - CONTEXT: The role of protein components of pancreatic secretions has been controversial in pancreatic stone formation. OBJECTIVE: To study the lithogenic role of pancreatic stone protein and lactoferrin in stone formation in chronic pancreatitis. PATIENTS: Pancreatic stones were collected from 13 patients with alcoholic (n=6) and nonalcoholic (n=7) chronic calcified pancreatitis. MAIN OUTCOME MEASURES: Pancreatic stone extracts were analyzed for pancreatic stone protein and lactoferrin using enzyme immunoassay. The localization of pancreatic stone protein immunoreactivity in the stone was observed using immunogold staining and scanning electron microscopy. RESULTS: Immunoreactivities for pancreatic stone protein were detected in the stones from all 13 patients with chronic calcified pancreatitis and for lactoferrin in the stones from five of the 13 patients. Pancreatic stone protein immunoreactivity distributed diffusely from the center to the periphery of the pancreatic stones. CONCLUSIONS: Involvement of pancreatic stone protein seems to be constant from the initial step of the stone formation to subsequent steps of the stone growth. However, pancreatic stone protein is only one of the precipitating proteins in pancreatic secretions such as lactoferrin, trypsinogen, etc. PMID- 11884768 TI - The radiologic spectrum of pulmonary Aspergillus infections. AB - Aspergillus infections may be categorized by specific radiographic patterns, the patient's immunologic status, and the presence or absence of preexisting structural lung disease. General patterns include invasive aspergillosis (both vascular and airway invasive varieties and acute tracheobronchitis), semiinvasive aspergillosis (including allergic bronchopulmonary aspergillosis and hypersensitivity pneumonitis), mycetoma, allergic aspergillosis, and obstructing bronchial aspergillosis. Knowledge of these various radiographic patterns as well as the immune derangements that accompany these infections may allow proper diagnosis. PMID- 11884769 TI - Solitary fibrous tumor of the pleura: MR appearance and enhancement pattern. AB - PURPOSE: The aim of the current study was to characterize the MR appearance of solitary fibrous tumor of the pleura (SFTP). METHOD: Twenty-two consecutive patients with histologically confirmed SFTP were retrospectively evaluated with MRI. RESULTS: Tumors demonstrated low signal intensity (n = 18, 82%), iso signal intensity (n = 2, 9%), and high signal intensity (n = 2, 9%) on T1-weighted images. On T2-weighted images, tumors were observed as low signal intensity (n = 10, 45%), high signal intensity (n = 2, 9%), and mixed signal intensity (n = 10, 45%). Solid component in the proliferation of tumor cells corresponded to low signal intensity in 12 tumors (55%) on T2-weighted images. On gadolinium-enhanced T1-weighted images, inhomogeneous enhancement (n = 18, 82%), nodular enhancement (n = 6, 27%), and homogeneous enhancement (n = 4, 18%) were observed. The enhanced lesion within the tumors had dense tumor tissue and dilated microvessels in the pathologic specimen. CONCLUSION: SFTP shows variable appearance and enhancement pattern on MRI according to morphologic tumor heterogeneity. PMID- 11884770 TI - Diffuse pulmonary neuroendocrine cell hyperplasia: radiologic and clinical features. AB - PURPOSE: The purpose of this study was to define the radiologic features of pulmonary neuroendocrine cell hyperplasia and correlate them with clinical findings. METHOD: Five women, ranging in age from 45 to 63 years, were diagnosed with pulmonary neuroendocrine cell hyperplasia. Two radiologists assessed the presence and extent of airway wall thickening, mosaic pattern, air trapping, ground-glass opacity, nodular opacity, and centrilobular opacity on high resolution CT. The CT findings were compared with physiologic data and histologic features. RESULTS: On CT scans, mosaic pattern was the predominant finding in all patients. The extent of mosaic pattern was correlated with the forced expiratory volume in 1 s/forced vital capacity ratio (r = 0.8508, p = 0.0317). Nodular lesions were noted in three patients. Airway walls were thickened in four patients. In one patient, ground-glass opacity and centrilobular opacity were also noted on high resolution CT. CONCLUSION: Pulmonary neuroendocrine cell hyperplasia is characterized by mosaic perfusion due to air trapping, airway wall thickening, and occasional small nodules on high resolution CT scans. The extent of mosaic perfusion correlates with physiologic evidence of airway obstruction. PMID- 11884771 TI - Lung tumors evaluated with FDG-PET and dynamic CT: the relationship between vascular density and glucose metabolism. AB - PURPOSE: The aim of the current study was to evaluate the relationship between FDG-PET and dynamic CT in lung tumors. METHOD: Forty consecutive patients with pulmonary tumors underwent whole-body FDG-PET and contrast-enhanced dynamic CT. The size of tumors was 2.6 +/- 0.2 cm (mean +/- SD) at the largest diameter. The standardized uptake value (SUV) of FDG-PET, peak attenuation (APA), and relative flow (RF) of dynamic CT were evaluated. All patients underwent surgery, and tissue samples were available to be studied. The intratumoral microvessel densities (MVDs) were counted and compared with the radiologic parameters. The duration between radiologic examinations and surgery was within 2 weeks in all patients. RESULTS: The mean SUV, APA, and RF of lung cancers were significantly higher than those of benign lesions (p < 0.05). The mean APA and RF of lung cancers correlated with mean SUV (APA : r = 0.665, p < 0.0001; RF: r = 0.848, p < 0.05) and mean MVD (APA: r = 0.801, p < 0.0001; RF: r = 0.723, p < 0.05). The mean SUV of lung cancers correlated with the mean MVD (r = 0.612, p < 0.001). No correlation was found between the mean APA, RF, SUV, and MVD in benign tumors. CONCLUSION: The APA and RF of dynamic CT correlated with the SUV of FDG-PET imaging in lung cancer. The APA and RF of dynamic CT as an index of blood pooling may be related to increased glucose metabolism in lung cancer. PMID- 11884772 TI - CT/fluoroscopy-guided transthoracic needle biopsy: sensitivity and complication rate in 98 procedures. AB - PURPOSE: The purpose of this study was to evaluate CT/fluoroscopy (CTF)-guided core needle biopsies (CNBs) in the thorax. METHOD: Ninety-eight biopsies were performed using a core biopsy needle (18G) with a reusable biopsy gun under CT/F. All results were compared to surgery plus histology or to clinical follow-up of >12 months. Sensitivity, specificity, and negative predictive value (NPV) were calculated. RESULTS: For pulmonary biopsies, sensitivity was 94%, specificity 100%, and NPV 73%; no significant correlation between the pneumothorax rate and the intrathoracic penetration depth was found. For biopsies of the mediastinum and pleura, sensitivities were 87 and 80%, respectively; specificity was 100% in both locations. A pneumothorax occurred in 21%, a pneumothorax requiring drainage in 2.0%. CONCLUSION: CT/F-guided CNB is a reliable method to obtain thoracic biopsies, with a complication rate of 2.0%. PMID- 11884773 TI - A pitfall of CT findings in peripheral lung adenocarcinoma. PMID- 11884774 TI - CT angiography of the subclavian artery: utility of curved planar reformations. AB - Despite advances in the diagnosis and treatment of peripheral vascular occlusive disease, an ever-aging population continues to provide scores of new cases requiring medical care. While traditional angiography has been the mainstay of diagnosis for many years, newer, less invasive techniques such as CT angiography with three-dimensional reformation are rapidly establishing themselves as first line diagnostic modalities. We present a case of severe left subclavian artery stenosis that demonstrates the utility of curved planar reformation in providing a concise visual summary of the pertinent anatomy and abnormalities. PMID- 11884775 TI - Image quality of three-dimensional electron beam coronary angiography. AB - PURPOSE: This study identifies reasons for poor image quality and nonassessability of coronary artery segments and compares results between early and late diastolic triggering on coronary electron beam angiography (EBA). METHOD: One hundred patients referred for EBA were studied. Contrast-enhanced transaxial coronary images were acquired using electrocardiographic (ECG) triggering and reconstructed three dimensionally using volume-rendering techniques. The image quality of coronary segments and image artifacts were analyzed statistically. RESULTS: Volume rendering failed in seven patients (7%) owing to cardiac and breathing motions. Image quality was the best with the left main (LM) and worst with the left circumflex (LCX) coronary arteries (p < 0.001). The image quality decreased systematically from proximal to distal within each coronary artery (p < 0.001). Forty percent R-R interval triggering on ECG was better than 80% for image quality. The nonassessable segments occurred in 3% of LM, 2, 8, and 5% of proximal, 24, 22, and 12% of mid, and 64, 45, and 20% of distal segments of the left anterior descending, LCX, and right coronary arteries, respectively (p < 0.05). CONCLUSION: The major limitations of coronary EBA were suboptimal spatial resolution and image artifacts. The image quality could be improved by using optimal ECG triggering. PMID- 11884776 TI - Risk of significant coronary artery disease as determined by CT measurement of the distribution of abdominal adipose tissue. AB - PURPOSE: The purpose of this work was to determine if CT measurement of the distribution of abdominal adipose tissue is reproducible between observers and is associated with patient risk of significant coronary artery disease. METHOD: We compared 11 male patients having abdominal CT who had a history of significant coronary artery disease and 9 male patients having abdominal CT without a history of coronary artery disease. Two observers, at the level of the umbilicus, independently measured the ratio of visceral adipose tissue (VAT) to total abdominal adipose tissue (TAT). VAT is equal to the sum total of intraperitoneal and retroperitoneal adipose tissue. TAT equals the sum total of visceral and subcutaneous adipose tissue. Measurements were made using a standard software package. RESULTS: The mean ratio of VAT to TAT was significantly different (p < 0.05) between patients with a history of coronary artery disease (mean = 0.51, SD = 0.10, range = 0.38-0.69) and without a history of coronary artery disease (mean = 0.40, SD = 0.12, range = 0.23-0.51). Agreement in measurements between observers was excellent (mean difference = 0.01, range = 0.00-0.03, intraclass correlation = 0.99). CONCLUSION: The measurement of the VAT/TAT ratio is highly reproducible between observers, and a high ratio is associated with patient risk of significant coronary artery disease. PMID- 11884777 TI - Contrast-enhanced MRI of the liver with mangafodipir trisodium: imaging technique and results. AB - Magnetic resonance (MR) contrast agents are now routinely used for detecting and characterizing focal liver lesions. Liver specific, hepatobiliary, MRI contrast agent mangafodipir trisodium (Mn-DPDP) is taken up by the functioning hepatocytes and excreted by the biliary system. Contrast uptake leads to persistent elevation of T1-weighted signal of normal liver parenchyma within 10 minutes of injection. Most tumors of non-hepatocellular origin typically are hypointense relative to enhanced liver parenchyma on T1 weighted images and are more conspicuous than on unenhanced images. Whereas, tumors of hepatocellular origin such as focal nodular hyperplasia (FNH), adenoma, and well-differentiated hepatocellular carcinomas (HCC) have been shown to accumulate Mn-DPDP, providing characterization information to discriminate hepatocellular from non-hepatocellular tumors. The purpose of this pictorial essay is to illustrate the appearance of various liver tumors on mangafodipir enhanced liver MR imaging. PMID- 11884778 TI - Intraductal papillary mucinous tumors of the pancreas: features with multimodality imaging. AB - The radiologic diagnosis of intraductal papillary mucinous tumor (IPMT) of the pancreas is important to establish because of its malignant potential and in order to determine the site of tumor origin and its extent. These pancreatic tumors are recognized more often now than previously because of the increasing use of imaging modalities such as computed tomography and magnetic resonance cholangiopancreatography. The main features of IPMT are given in this pictorial essay, with illustrations provided from a series of 50 patients with a surgically proven diagnosis of IPMT. PMID- 11884779 TI - MR findings of renal cortical necrosis. AB - PURPOSE: Evaluating the MR findings of renal cortical necrosis was the purpose of this study. METHOD: Eight series of T1-/T2-weighted (n = 8) and contrast-enhanced T1-weighted (n = 4) MR images in six patients with renal cortical necrosis diagnosed by renal biopsy (n = 4) or on clinical grounds (n = 2) were reviewed. In those who had follow-up MRI (n = 2) or comparable CT (n = 3), interval changes of MR findings and comparison with CT images were done. RESULTS: Swollen kidney with dark signal intensity rim involving the inner cortex and column of Bertin was noted on T2-and T1-weighted images. It was more conspicuous on T2-weighted images. The lesion did not enhance and was differentiated from uninvolved renal parenchyma. In the follow-up MRI, thickened dark signal intensity was more prominent and proved to be calcification or fibrosis. CONCLUSION: MRI, especially T2-weighted and contrast-enhanced T1-weighted imaging, was helpful in evaluating renal cortical necrosis. PMID- 11884780 TI - MR cholangiography in the evaluation of hepatic and biliary abnormalities in autosomal dominant polycystic kidney disease: study of 93 patients. AB - PURPOSE: The purpose of this work was to conduct an MR cholangiography study of hepatobiliary abnormalities in autosomal dominant polycystic kidney disease (ADPKD) and to correlate these abnormalities with the risk of infection. METHOD: Cystic and intrahepatic bile duct (IHBD) abnormalities identified by MR cholangiography in 93 ADPKD patients were studied retrospectively. A blind study of liver function tests, renal insufficiency, and infectious episodes was also carried out. Correlations among these data were looked for using univariate analysis. RESULTS: Intrahepatic cysts were present in 84 of 93 patients and peribiliary cysts in 59 of 90 patients. IHBDs were abnormal in 25 of 90 patients (tubular dilatation in 15 cases and bead-like dilatation in 16). IHBD abnormalities were correlated with hepatobiliary infections (p = 0.0012), gamma glutamyltranspeptidase elevation (p = 0.018), and terminal renal failure (p = 0.006). CONCLUSION: This study confirms the complexity of hepatobiliary involvement in ADPKD. Various types of cystic lesions can arise, and patients with IHBD abnormalities are at increased risk for hepatobiliary infection. PMID- 11884781 TI - Hepatic blood flow measurements with arterial and portal blood flow mapping in the human liver by means of xenon CT. AB - PURPOSE: The purpose of this work was to quantify arterial and portal blood flows in the human liver and to create blood flow maps by means of xenon CT. METHOD: Mathematical procedures were developed based on a simplified model having two tissue components: liver tissue and portal organ tissue. Xe-CT studies were performed on 10 healthy volunteers (ages 33.4 +/- 9.8 years), a patient with hepatocellular carcinoma (HCC), and a liver transplant recipient. RESULTS: Arterial and portal blood flows for the healthy subjects were 36.7 +/- 5.2 and 65.2 +/- 22.0 ml/100 ml/min. In the HCC patient, arterial blood flow was shown to be dominant in the tumoral area. From the results of the liver recipient, it was demonstrated that obtaining lambda values is important for proper evaluation of blood flows. CONCLUSION: Xe-CT can provide substantial information on hepatic blood flow quantitatively and visually with separation of arterial and portal components. PMID- 11884782 TI - Diffusion-weighted echo planar imaging of ovarian tumors: is it useful to measure apparent diffusion coefficients? AB - PURPOSE: Our goal was to test the hypothesis, as previously reported in other studies, that apparent diffusion coefficients (ADCs) provide specific information to diagnose ovarian tumors, especially to discriminate between benign and malignant lesions. METHOD: T1-and T2-weighted spin echo imaging and diffusion weighted echo planar imaging were performed in 31 women with 61 cystic components of ovarian tumors. RESULTS: The lesions that showed typical watery intensity, hypointensity in T1-weighted imaging, and hyperintensity in T2-weighted imaging had similar ADCs, ranging from 1.54 to 1.84 x 10(-3) mm2 /s. The lesions that showed signal intensity different from typical watery intensity in conventional MRI tended to have low ADCs. In endometrial cysts, the mean ADC of the subgroup that showed typical watery intensity was higher than that of other subgroups. CONCLUSION: With conventional MRI, a tendency of ADCs could be predicted. ADCs may not provide additional information, especially to discriminate benign from malignant lesions. PMID- 11884783 TI - MR findings of Erdheim-Chester disease. AB - Lipoid granulomatosis (Erdheim-Chester disease) is a rare but distinct form of histiocytosis. This disease has characteristic radiologic findings involving the musculoskeletal system that are critical to the diagnosis: symmetric sclerosis of the metaphysis and diaphysis of long bones with relative sparing of the epiphysis as depicted on conventional radiography. However, it is a systemic disease that involves multiple organ systems. This pictorial essay is of a single patient imaged over multiple years, using various pulse sequences with both low and high field strength MR scanners. It depicts many of the characteristic findings encountered in this rare systemic disorder. PMID- 11884784 TI - Proteus syndrome. AB - Proteus syndrome is a rarely described dysplasia syndrome of the group of congenital hamartomas that arises from mosaic mutation. An extraordinary case history including imaging studies will be reported. This 17-year-old girl suffered from cachexia, lifelong chronic obstipation, different dysplasias, and lipomatous tumor-like lesions. The following findings were marked: macrodactyly, nevi, hemihypertrophy, aggressive lipomatosis, hemangiomas of the spleen, and skull and cerebral malformations. Additionally, an intestinal affection with fatty wall thickening was detected. In contrast to reports in the literature describing a reduced lifespan with a mean of few years, our patient is still alive. The treatment should take a palliative symptomatic approach considering the clinical situation. PMID- 11884785 TI - Thin section MR study of the basal ganglia in the differential diagnosis between striatonigral degeneration and Parkinson disease. AB - PURPOSE: Signal abnormalities within the putamen in MRI have been related to tissue degeneration in the striatonigral variant of multiple system atrophy (MSA P). While previous work demonstrated the high specificity of these MR findings, sensitivity rates were unsatisfactory. We evaluated the specificity and sensitivity of an acquisition protocol using thin section MRI to differentiate MSA-P from Parkinson disease (PD). METHOD: Axial 3-mm-thick conventional T2 and proton density spin echo images at the level of basal ganglia were acquired at 1.5 T in 24 patients with MSA-P and 27 patients with PD. RESULTS: We found an abnormal putaminal T2 hypointensity in 21 of 24 MSA-P patients (87.5% sensitivity) and a proton density hyperintensity in 20 of 24 MSA-P patients (83.3% sensitivity). Three among 27 PD patients had an abnormal putaminal T2 hypointensity (88.8% specificity) and there were no proton density abnormalities (100% specificity). CONCLUSION: Our thin section conventional spin echo protocol showed a substantial increase in MR sensitivity compared with previous reports. We believe that a better depiction of even mild signs of degeneration in the putamen may allow a more widespread use of this technique in the differential diagnosis of parkinsonisms. PMID- 11884786 TI - Evaluation of cerebral perfusion parameters measured by perfusion CT in chronic cerebral ischemia: comparison with xenon CT. AB - PURPOSE: The purpose of this work was to evaluate the usefulness of perfusion CT in the evaluation of patients with chronic cerebral ischemia by comparing it with xenon CT (Xe-CT). METHOD: Cerebral blood flow (CBF) of perfusion CT (CBFper) and time to peak (TTP) were compared with the CBF of Xe-CT (CBFxe) in 18 patients. Cerebral blood volume (CBV) was compared with cerebral vascular reserve (CVR) in 10 of 18 patients who underwent pre- and postacetazolamide Xe-CT. RESULTS: CBFper and TTP demonstrated a high correlation with CBFxe in relative values by side-to side comparisons (r = 0.743, p < 0.0001 and r = -0.760, p < 0.0001, respectively). There was a negative correlation between relative CBV and relative CVR (r = -0.637, p = 0.0025). Visually, territories with delayed TTP corresponded well to those of decreased CBFxe, but these territories tended to be larger in TTP maps. CONCLUSION: Perfusion CT is a useful tool to evaluate chronic hemodynamic disturbance and can be an alternative method for those using acetazolamide challenge. PMID- 11884787 TI - Tumor staging of laryngeal and hypopharyngeal carcinomas with functional spiral CT: comparison with nonfunctional CT, histopathology, and microlaryngoscopy. AB - PURPOSE: The purpose of this work was to compare nonfunctional and functional spiral CT in the tumor (T) staging of laryngeal and hypopharyngeal tumors and to correlate the CT results with microlaryngoscopy and postoperative pathology. METHOD: Twenty-six patients (3 women, 23 men) with clinically suspected laryngeal and hypopharyngeal tumors underwent both nonfunctional CT during quiet breathing and functional spiral CT during either a modified Valsalva (n = 19) or E phonation (n = 7) maneuver. CT slice thickness was 3 mm, table feed was 3 mm, and 40-80 ml of intravenous contrast material was administered at a flow of 1.5 ml/s. T stages as determined by nonfunctional and functional CT were compared and correlated with postoperative pathology or microlaryngoscopy. RESULTS: The T stages determined with functional CT were better correlated with postoperative pathology (rS = 0.88, p = 0.001) and microlaryngoscopy (rS = 0.77, p = 0.008) than T stages determined with nonfunctional CT (rS = 0.80, p = 0.001; and rS = 0.51, p = 0.13, respectively). Twelve of 26 patients (46%) had a lower T stage on functional than on nonfunctional CT. In 14 of 26 patients (54%), the T stage was identical with both modalities. In no patients was the T stage increased by functional CT. CONCLUSION: Functional CT appears to be more accurate than nonfunctional CT in the T staging of laryngeal and hypopharyngeal carcinomas. Functional CT also results in lower T stages than nonfunctional CT in a substantial number of patients. PMID- 11884788 TI - Sinus pericranii: demonstration using three-dimensional surface shading. PMID- 11884789 TI - Contrast-enhanced CT with saline flush technique using two automated injectors: how much contrast medium does it save? AB - PURPOSE: The purpose of this work was to investigate the volume of contrast medium saved by the saline flush technique. METHOD: Thoracic helical CT was performed by injecting 75 ml of contrast material (Ioversol 320) only (n = 25; Group A) or 75 ml of contrast medium pushed with 6 ml (n = 25; Group B), 12 ml (n = 25; Group C), 25 ml (n = 25; Group D), or 50 ml (n = 25; Group E) of saline at a rate of 2 ml/s. The aortic CT numbers were measured from 30 to 55 s after the beginning of injection. We compared the time to peak aortic enhancement (TPAE) among Groups A-E. RESULTS: The TPAEs of the Groups A-E were 42.6, 45.2, 48.6, 48.4, and 48.3 s, respectively, and there was a statistically significant difference among them (p < 0.0001, analysis of variance). Post hoc test revealed statistically significant differences in TPAE between Groups A and B, C, D, and E and between Groups B and C, D, and E but no differences among Groups C, D, and E. CONCLUSION: The saline flush technique prolongs TPAE by 6 s and saves 12 ml of contrast medium. PMID- 11884790 TI - Image-guided coaxial fine needle aspiration biopsy with a side-exiting guide. AB - PURPOSE: The aim of this study was to evaluate the diagnostic accuracy and complication rates of a side-exiting coaxial needle system for fine needle aspiration (FNA) biopsies. METHOD: Between 1995 and 1998, 127 nonconsecutive biopsies were performed on 122 patients (74 males, 48 females). CT guidance was used in 111, ultrasound guidance was used in 14, and both were used in 2 biopsies. Patient history, biopsy site, needle performance, complications, and cytology results were recorded. RESULTS: Diagnostic rate and accuracy were 92.9 and 99.2%, respectively. There were minor complications from 14 biopsies, and all of them arose from chest biopsies: pneumothorax in 13 of 47 and hemoptysis in 1 of 47. There were no major complications. CONCLUSION: The side-exiting coaxial needle system is a safe and effective alternative to the conventional end-exiting coaxial needle system for performance of image-guided FNA biopsies. PMID- 11884791 TI - Morphometric analysis of cortical sulci using parametric ribbons: a study of the central sulcus. AB - Interhemispheric and gender differences of the central sulcus were examined via a parametric ribbon approach. The central sulcus was found to be deeper and larger in the right nondominant hemisphere than in the left dominant hemisphere, both in males and in females. Based on its pattern, that asymmetry could be attributed to increased connectivity between motor and somatosensory cortex, facilitating fine movement, which could constrain the in-depth growth of the central sulcus. Position asymmetries were also found, which might be explained by a relative larger parietal association cortex in men but not in women. PMID- 11884792 TI - Reduction of electronic noise from radiofrequency generator during radiofrequency ablation in interventional MRI. AB - MRI has been used increasingly in the recent past for the guidance and monitoring of minimally invasive interventional procedures, using typically radiofrequency (RF) and laser energy, cryoablation, and percutaneous ethanol. RF energy has been used over the last 30 years for the ablation of tissues. Its use in conjunction with MRI for monitoring is limited, however, because of the electronic noise produced by the RF generators, which can significantly deteriorate image quality. The objective of this work was to devise methods by which this noise can be reduced to an acceptable level to allow simultaneous acquisition of MR images for monitoring purposes with the application of RF energy. Three different methods of noise reduction were investigated in a 0.2 T MR scanner: filtration using external hardware circuitry, MR scanner software-controlled filtration, and keyholing. The last two methods were unable by themselves to suppress the noise to an acceptable degree. Hardware filtration, however, provides excellent suppression of RF noise and is able to withstand up to 12 W of RF energy. When all the three approaches are combined, significant reduction of RF noise is achieved. The feasibility of creating an RF lesion of about 1.2 cm diameter in vivo in a porcine model simultaneously with temperature-sensitive MRI with adequate noise suppression is demonstrated. PMID- 11884793 TI - A computed tomographic guide to endoscopic sinus surgery: axial and coronal views. AB - SUMMARY: The purpose of this article is to correlate endoscopic surgical procedures in the sinuses with computer tomographic (CT) scans. Twelve commonly performed procedures were color-coded on a set of normal coronal and axial CT scans. The illustrated procedures are uncinectomy/antrostomy, anterior ethmoidectomy, posterior ethmoidectomy, sphenoidotomy, partial inferior turbinectomy, partial middle turbinectomy, septoplasty, frontal recess approach, dacrocystorhinostomy, orbital decompression, optic nerve decompression, and medial maxillectomy. Drainage of a medial orbital abscess is discussed but not illustrated. A brief description of the indications, surgical approach, and complications of each procedure is also provided. PMID- 11884794 TI - The SOOF lift: its role in correcting midfacial and lower facial asymmetry in patients with partial facial palsy. AB - Subperiosteal face lifting has gained wide acceptance in aesthetic surgical practice. It may also have a role to play in patients with partial facial palsy. These patients demonstrate poor static position of the mouth but maintain some degree of facial movement. This study examined the role of subperiosteal facial suspension as an alternative treatment modality in this patient group. In this series, five patients with varying degrees of partial facial palsy underwent subperiosteal face lifting, including sub-orbicularis oculi fat elevation via a temporal, lower lid, and buccal approach, thereby mobilizing and elevating and suspending the zygomaticus major and levator labii superioris muscles on the facial skeleton. An attempt was made to categorize the patients according to overall House-Brackmann score. It was not possible to precisely classify the patients by this method, although the approximate scores were two patients scoring 3, two patients scoring 4, and one patient scoring 5. To overcome inconsistencies with this method, the degree of static and dynamic asymmetry of the mouth and also the excursion of the mouth were graded separately. Four patients with mild to moderate dynamic and static asymmetry (House-Brackmann score of approximately 3 and 4) who maintained excellent or good excursion of the mouth achieved excellent or good results. One patient with poor excursion and severe partial facial palsy (House-Brackmann score of 5) was improved but remained markedly asymmetric (follow-up, 4 months to 1 year). Subperiosteal face lifting is a useful therapeutic modality for management of selected patients with mild partial facial palsy. These patients demonstrate asymmetric static position but maintain some degree of muscle excursion. Patients with severe facial palsies with poor muscle excursion continue to require muscle transfer or sling procedures. The authors hope that long-term follow-up will confirm the sustained effect of midfacial suspension in this selected patient group. PMID- 11884796 TI - Facial artery in the upper lip and nose: anatomy and a clinical application. AB - Twenty-five facial arteries were examined radiographically in 19 fresh cadavers that had been injected systemically with a lead oxide-gelatin mixture. Major branches of the facial artery in the upper lip and nose were investigated, and the anatomical variations were classified into three types on the basis of the anatomy of the lateral nasal artery, which was determined as an artery running toward the alar base. In 22 cases (88 percent), the facial artery bifurcated into the lateral nasal artery and superior labial artery at the angle of the mouth. In two cases (8 percent), the facial artery became an angular artery after branching off into the superior labial artery and the lateral nasal artery sequentially. In one case (4 percent), the facial artery became an angular artery after branching off into the superior labial artery, and the lateral nasal artery then branched off from the superior labial artery. Branches from the lateral nasal and superior labial arteries were observed stereographically. Vascular anastomoses between those branches were created in the upper lip, columella base, and nasal tip, and an intimate vascular network was formed. With a vascular network in the mucosa of the upper lip, a bilobed upper-lip flap was created for a clinical case with a full-thickness defect of the ala. PMID- 11884799 TI - Endoscopic endonasal reconstruction of blowout fractures of the medial orbital walls. AB - High-resolution endoscopes and the advent of endoscopic instruments for sinus surgery currently provide surgeons with excellent endonasal visualization and access to the medial orbital walls. The purpose of this study was to demonstrate the reduction of medial orbital wall fractures through an endonasal endoscopic approach that allows the repair of the medial orbital wall fractures without an external incision. This study was a retrospective analysis of 16 patients who underwent surgical repair of medial orbital wall fractures from March of 1997 to May of 1998. The 11 male and five female patients ranged in age from 16 to 54 years (mean, 30.5 years). These patients had undergone primary reduction of medial orbital wall fractures and were observed for at least 12 months after surgery. There were no intraoperative or postoperative complications. Fifteen of 16 patients showed a complete improvement of their symptoms. One patient showed persistent diplopia, which was well managed by prisms. Endoscopic reduction of medial orbital wall fracture using an endonasal approach seems to produce good results and definite cosmetic advantages. PMID- 11884798 TI - Cranial reconstruction with computer-generated hard-tissue replacement patient matched implants: indications, surgical technique, and long-term follow-up. AB - The aim of this clinical study was to evaluate the effectiveness and safety of using computer-generated alloplastic (hard-tissue replacement) implants for the reconstruction of large defects of the upper craniofacial region. Fourteen patients who had large (> 150 cm2) preexisting defects of the cranium or cranio orbital region underwent surgical reconstruction. Preoperatively, a three dimensional computed tomographic scan was obtained from which an anatomic model was fabricated. The defect in the model was then used to create an alloplastic (hard tissue-replacement polymer) implant for reconstruction and surgical placement. At the time of surgery, the implant was secured into position with either metal or resorbable fixation. In cases where the frontal sinus was in proximity to the implant, the frontal sinus was either cranialized and covered with a pericranial flap or obliterated with hydroxyapatite cement. In cases that had been previously irradiated or infected, wide bony debridement and coverage with a vascularized muscle was initially performed, followed by implant reconstruction 6 months later. All implants fit easily into the bone defects, and only four (29 percent) required some minor adjustments to complete the fit. All patients healed uneventfully. With a minimum of 1 year follow-up (average, 3 years) in all cases, excellent contours have been maintained and all patients have remained infection-free. In large cranial defects, custom implants fabricated from porous, hydrophilic hard-tissue replacement polymer provide an exacting anatomic fit and a solid stable reconstruction. This method of reconstruction in these defects is rapid and exact, and significantly reduces operative time. Critical attention must be paid, however, to management of the frontal sinus and preexisting bone infection and the quality of the overlying soft-tissue cover. PMID- 11884800 TI - Porous polyethylene implants in orbital floor reconstruction. AB - The purpose of this article is to present the authors' experience with the use of porous polyethylene ultrathin sheets for orbital floor reconstruction. Thirty-two patients with orbital floor fractures were treated with porous polyethylene ultrathin sheets. Sixteen cases corresponded to orbitozygomatic fractures, 11 cases corresponded to pure orbital floor fractures, and five corresponded to panfacial fractures. The subciliary approach was used in 15 patients and the transconjunctival approach in nine; another three patients were operated on through a preexisting eyebrow wound, two were operated on with a subtarsal approach, two were operated on through an eyebrow extension of a facial wound, and one patient was operated on through the facial wound. Intraoperatively, all patients received a prophylactic dose of intravenous antibiotics. Postoperatively, 24 patients received amoxicillin clavulanate for 5 to 7 days, two patients received clindamycin, and six patients received no antibiotics. Enophthalmos was corrected in 15 of 24 patients (62.5 percent), and hypoglobus in nine of 11 (82 percent). Diplopia was resolved in 25 of 28 patients (89.3 percent) with preoperative impairment. Extrinsic eye movement impairment was resolved in 25 of 27 patients (92.6 percent). A preoperative visual acuity deficit was present in four patients (12.5 percent) and was resolved in one (from 20/100 to 20/20). Visual acuity improved in one patient (from 20/60 to 20/30). In the other two patients, visual acuity remained altered (from 20/30 to 20/30). One patient (3.1 percent) suffered blindness induced by surgery. Nine of 26 patients (34.6 percent) had residual infraorbital nerve hypesthesia and five (19.2 percent) had residual paresthesias. Postoperatively, epiphora was present in six patients (18.8 percent) and ectropion in five (15.6 percent). Although there was no statistical significance between the surgical approach and the presence of epiphora (p = 0.211) and ectropion (p = 0.422), patients who were treated using the transconjunctival approach suffered reduced ectropion (0 percent) compared with patients treated using the subciliary approach (20 percent). However, patients treated using the transconjunctival approach suffered increased epiphora (22.2 percent) compared with those treated with the subciliary approach (13.3 percent). There were four cases (12.5 percent) of postoperative facial infections. Two of these cases were resolved with systemic antibiotics, one was resolved with bone sequestrum resection, and one patient needed removal of the implant. Orbital infections were related in all cases to titanium osteosynthesis miniplates or skull bone graft. When comparing patients who were treated with and without antibiotics, no statistical differences (p = 0.958) were found relative to the presence of infections. Correction of hypoglobus is technically easier than enophthalmos, because enophthalmic correction requires a wide, deep subperiosteal dissection and implant positioning, posterior to the equator of the globe, with the inherent risk of orbital apex injury. PMID- 11884802 TI - Regeneration of intraoral defects after tumor resection with a bioengineered human dermal replacement (Dermagraft). AB - The experiences of seven patients with squamous cell carcinomas of the oral cavity who underwent reconstruction with a bioengineered human dermal replacement (Dermagraft) are examined. The human dermal replacement consists of fibroblasts seeded onto a three-dimensional polymer scaffold to create a living dermal structure. In this setting, the fibroblasts secrete a mixture of growth factors and matrix proteins in physiological concentration that is essential for wound healing and epithelization. The fibroblast tissue remains metabolically active after cryopreservation and can be used as an off-the-shelf tissue to cover medium sized defects and avoid donor-site morbidity. In the first series of patients treated with this tissue, defect closure was achieved without functional problems, allowing optimal postoperative monitoring for tumor recurrence. PMID- 11884804 TI - Mandibular distraction in neonates: a strategy to avoid tracheostomy. AB - Over the past 5 years, the authors developed an application of mandibular distraction osteogenesis to eliminate existing tracheostomy. That experience led the authors to attempt mandibular distraction osteogenesis in neonates as an alternative before tracheostomy. Success with this approach using supporting objective airway measurements has been reported previously. This report includes six neonates diagnosed with Pierre Robin sequence. Of the six, five neonates ranging in age from 6 to 26 days (mean, 14.5 days) were treated by the authors with mandibular distraction over a 22-month period. The sixth neonate was treated with tracheostomy, because of other airway abnormalities. Findings included retrognathia, glossoptosis, incomplete cleft palate, and airway obstruction in each patient. Birth weights ranged from 2.8 to 3.2 kg. All patients were unable to control their airway during feeding, as evidenced by repeated episodes of choking and obstruction. Resting oxygen saturations were in the 70 to 80 percent range in all patients, with further deterioration during attempted feeding. Bronchoscopy was performed in all patients under anesthesia before distraction. Recurrent near-complete and intermittent complete airway obstruction were present in all patients at the level of the tongue base. There was a consensus by a pediatric intensivist, a pediatric anesthesiologist, and a pediatric otolaryngologist in all cases. Each patient met all criteria requiring ventilation for life support. Tracheostomy would be required if mandibular distraction osteogenesis was not performed, or if it failed. Patients with other airway abnormalities were not considered for treatment. Maxillomandibular disharmony measured at the midline ranged from 8 to 15 mm (mean, 11.2 mm). Active distraction was performed at the rate of 1 to 2 mm a day, with a consolidation period of 4 weeks. Total time of treatment was less than 6 weeks in all cases. All patients were extubated by the completion of active distraction. Distraction distance ranged from 8 to 15 mm (mean, 12.4 mm). All patients were discharged to home on apnea monitors, the use of which was discontinued after 90 days with no further apneic events. Weight gains met or exceeded the average 500 g a month after distraction. Bronchoscopy at the time of distractor removal showed correction of airway obstruction at the tongue base. Radiographs showed bilateral ossification of the distraction sites. Tracheostomy was avoided in all cases selected for treatment by distraction. Patient follow-up range was 9 to 22 months. In selected Pierre Robin sequence patients with tongue base airway obstruction, mandibular distraction osteogenesis can successfully avoid the need for and the associated mortality and morbidity of indwelling tracheostomy. PMID- 11884806 TI - Early nonsurgical correction of congenital auricular deformities. AB - Congenital auricular deformation is not an uncommon phenomenon, and it may cause substantial annoyance to the growing child. Many parents of affected children will seek surgical correction of the deformed auricles. The variety of techniques that have been described for the correction of this anomaly suggests that none has been considered satisfying. The consequent possible surgical complications should also be considered, when the surgical procedure can be replaced by an effective conservative treatment. The authors describe their experience using early splinting for congenital auricular deformities. Fifty-two newborn infants with lop, prominent, Stahl's, and constricted ears referred to us by the neonatal department staff were enrolled in this study. Putty Soft, a vinyl polysiloxane impression material, has been used for early molding of the auricles. Surgical tapes were used for the fixation of the mold and to fix the auricle to the scalp. The results were evaluated by one of the parents and by a layperson (medical student) 6 months after completion of the procedure. The above-described early splinting procedure was applied onto 92 auricles of 52 newborn infants aged 1 to 10 days, mostly around day 3. The mean treatment time was 6.8 weeks. All treated auricles were improved, 87 percent were rated as excellent improvement, and there were no complications related to the treatment. The authors conclude that early splinting of deformed auricles should be offered to parents of affected children, and the awareness of this procedure by neonatologists, pediatricians, and nursery staff should be increased. PMID- 11884808 TI - Stability of dental implants in microvascular osseous transplants. AB - Microvascular iliac crest and scapula transplants have been used in reconstruction of the lower jaw following tumor surgery. It has only been with the insertion of dental implants that a satisfactory prosthetic rehabilitation of the patient has been achieved. For this study, a follow-up of 38 patients with lower jaw tumors was carried out. The patients had been treated with partial resection of the lower jaw and neck dissection with microvascular iliac crest transplants (n = 20) or microvascular scapula transplants (n = 18); this was followed with dental implants (n = 143) in the region of the transplants or the local lower jaw. One hundred thirty-nine of the 143 dental implants were loaded by prosthetic superstructures. In all patients, the implant situation was evaluated on average 2 years 5 months after implantation. Periotest values, periimplant probing depths, and contact bleeding were registered, and the extent of periimplant bone loss was defined radiographically. The clinical situation in the region of the implant was compared for both types of implants and also with the nonresected lower jaw. The average Periotest values were within the normal range for all groups. In one scapula implant, however, a better average of Periotesting, -3.3, was found compared with implants of the iliac crest with Periotest values of -0.7. A measurement of -2.1 was found for the local lower jaw, similar to that of scapula implants. Pathologic probing depths were found for all three compared groups. The radiographically determined vertical loss of bone was the same for all three groups, on average 1 mm at 27 months postoperatively. The highest incidence of sulcus bleeding was found in the scapula implant group. Thus, it can be stated that the scapula transplants provide a similar transplant site to local lower jaw bone, whereas implants in iliac crest transplants show lesser bony stability. Periimplant soft-tissue conditions are worse for both types of transplants compared with local tissue of the lower jaw. PMID- 11884810 TI - Distraction osteogenesis of costochondral bone grafts in the mandible. AB - Costochondral grafting for reconstruction of the Pruzansky type III mandible has given variable results. Lengthening of the rib graft by means of distraction had been advocated when subsequent growth of the grafted mandible is inadequate. This retrospective study reviews a series of patients with mandibular costochondral grafts who underwent subsequent distraction osteogenesis of the graft. A retrospective review identified two patient groups: group 1 consisted of individuals (n = 9) who underwent costochondral rib grafting of the mandible followed by distraction osteogenesis several months later at a rate of 1 mm/day. Group 2 consisted of patients with Pruzansky type II mandibles who had distraction osteogenesis without prior rib grafting (n = 9). The biomechanical parameters, orthodontic treatment regimens, and complications were examined versus patient age and quality of the rib graft. Distraction osteogenesis was successfully performed in six of the rib graft patients (group 1) and in all of the group 2 individuals. On the basis of the Haminishi scale, the computed tomographic scan appearance of the regenerate was classified as "standard or external" in six of the group 1 patients and as either "agenetic" or "pillar" (fibrous union) in the remaining three patients. In group 1, the average device was expanded 23 mm (range, 20 to 30 mm). Group 2 mandibular distraction results were all classified as either standard or external, and there was an average device expansion of 22.4 mm (range, 16 to 30 mm). The length of consolidation averaged 12.6 weeks in group 1, compared with 8.5 weeks in the traditional mandibular distraction patients (group 2). The mean shift of the dental midline to the contralateral side was 2.5 mm in group 1 versus 4.0 mm in group 2. Complex multiplanar and transport distractions were successfully performed on grafts of adequate bony volume. All four patients in group 1 with tracheostomies were successfully decannulated after consolidation. Rib graft distraction complications included pin tract infections in two patients, hardware failure with premature pin pullout in one patient, and evidence of fibrous nonunions in three young patients with single, diminutive rib grafts. In group 2, there were no distraction failures. Distraction osteogenesis can be successfully performed on costochondral rib grafts of the mandible; however, the complication rate is higher than in non-rib-graft patients. Performing the technique on older, more cooperative individuals seems to reduce this risk. In addition, placement of a double rib graft or an iliac bone graft of sufficient volume to create a neomandible with greater bone stock is an absolute requirement to decrease the risk of fibrous nonunion and provide a bone base of sufficient size for retention of the distraction device and manipulation of the regenerate. PMID- 11884812 TI - Prefabricated superficial temporal fascia flap combined with a submental flap in noma surgery. AB - The authors report their experience with a new procedure: the combination of a prefabricated superficial temporal fascia flap and a submental flap performed in an African hospital on five patients with cheek deformities caused by noma. The prefabricated superficial temporal fascia flap makes the inner lining of the cheek, which is anchored on the peripheral scar tissue. The submental flap is released during the second operation and makes the outer lining. The main advantages are the excellent aesthetic color of this last flap and the short distance between the donor site and the recipient site. Moreover, the submental flap is positioned in a single operation (when the outer-lining reconstruction is performed with a deltopectoralis flap, a third operation is necessary to cut the pedicle). None of the flaps failed, and the functional results were good. The prefabricated superficial temporal fascia flap and submental flap are versatile and reliable flaps, with reasonably long vascular pedicles, that can be used successfully, even under suboptimal conditions in weak patients with huge defects of the face. PMID- 11884814 TI - The relationship of facial two-point discrimination to applied force under clinical test conditions. AB - When an interside comparison is hampered, for example, in cases of bilateral trauma, normal threshold values of two-point discrimination from healthy subjects might be used to delineate abnormal from normal sensory function in patients. To determine threshold pin distances, two devices, the Disk-Criminator and the Aesthesiometer, have often been applied in a clinical setting. Because these devices are hand-operated, the force of applying a device might vary considerably. The general applicability of normal threshold values from the literature may therefore be questioned. Five subjects participated in experiments with two observers, in which a hand-operated device with a constant pin distance (5 or 10 mm) was pushed on a facial site, until the point at which blanching of the skin started, and the applied force was recorded. To that end, the devices were modified by providing them with force transducers. These recordings revealed a considerable variation in force variables (level, duration, and rate). Significant differences in mean force level, duration, and rate occurred, particularly between devices, pin distances, and/or sites (cheek, upper and lower lips, and mental region) and also in mean duration between observers (p < 0.01 to 0.001, analysis of variance). However, the observed force levels were always at an extremely supra-threshold stimulus intensity. The threshold pin distances in subsequent experiments (four subjects, the two devices and two sites: cheek and mental region) were therefore almost invariant to the difference in the extreme low and high force levels that were applied, using acoustic feedback on the force signal. Furthermore, these thresholds were also similar when one or two observers performed repeated measures on groups of 15 to 18 subjects under the influence of a usual variation of force level, using an interval of at least 1 week and both nonmodified devices. Because of invariance, normal values of threshold pin distance are generally applicable to any well-trained observer and are related to the density of afferent nerve fibers. In contrast, recently reported force thresholds determined at a constant pin distance might not be related to fiber density only. The findings regarding dependency on site and pin distance of the force level suggest that force thresholds will also be related to tissue stiffness and to the extent to which a pair of pins co-operate mechanically. PMID- 11884816 TI - Low-dose propofol infusion for sedation during local anesthesia. AB - The safety and efficacy of lose-dose propofol for sedation were investigated on 90 consenting patients who had undergone surgical procedures with local anesthesia. After being premedicated with intravenous midazolam 0.05 mg.kg(-1), all patients were randomly divided into two groups and received intravenously either a loading dose of propofol 0.8 mg.kg(-1) followed by a continuous infusion of propofol 30 microg.kg(-1)min(-1) (propofol group) or an equivalent volume of saline (placebo group) during operation. Study groups were compared with respect to the level of sedation, hemodynamic variables, oxygen saturation, and the incidence of intraoperative side effects. In addition, the discharge time and the satisfaction of both patients and surgeons with this sedative technique were assessed. Propofol reduced patients' discomfort and lowered their arterial pressure and heart rate during the infiltration of local anesthetics. It also promoted an adequate level of sedation without clinically significant oxygen desaturation in the intraoperative period. Surgeons and patients in the propofol group showed a higher level of satisfaction than those in the placebo group. There was no significant difference between the two groups with regard to the incidence of adverse effects and the discharge time. In conclusion, it was found that the use of low-dose propofol infusion was a safe and effective sedative technique for local anesthesia. PMID- 11884817 TI - Cutaneous leiomyosarcoma. AB - Cutaneous leiomyosarcoma is a rare soft-tissue sarcoma with negligible metastatic potential, but local recurrence rates after surgical excision have ranged from 14 percent to 42 percent. Unlike other sarcomas, guidelines for the optimal surgical excision margin of cutaneous leiomyosarcoma are not clearly defined in the existing literature. A review of local experience with this condition revealed eight patients over 12 years, none of whom developed local recurrence or distant metastases. This is despite poor prognostic factors in seven patients and excision margins ranging from 1 to 27 mm. These findings are compared with previously published data, and conclusions are drawn based on analysis of the collective results. Complete surgical excision with a narrow margin is recommended, and patients should be observed for a minimum of 5 years after surgery. PMID- 11884818 TI - The role of reduction mammaplasty in reconstructing partial mastectomy defects. AB - The management of breast tumors in women with macromastia can be challenging. Reconstructive options are limited and breast conservation therapy is often not indicated or results in poor cosmetic outcomes. The purpose of this report was to present a series of women with macromastia who underwent simultaneous reconstruction of a partial mastectomy defect with bilateral reduction mammaplasty. A retrospective review was performed and included all women who underwent partial mastectomy with simultaneous reduction mammaplasty. Data points included patient demographics, preoperative assessment, operative intervention, adjuvant treatment, and outcomes. Twenty women were included in the series (mean age, 43 years; range, 11 to 72 years) with an average body mass index of 32.6 (range, 24.9 to 44.1). Tissue diagnosis was ductal carcinoma (n = 8), ductal carcinoma in situ (n = 6), fibroadenoma (n = 4), and benign breast tissue (n = 2). The various reduction mammaplasty techniques were documented with regard to tumor size and location. The superior medial and inferior pedicles seemed to be the most versatile techniques. One patient required completion mastectomy with autologous tissue reconstruction given positive margins. All patients were disease-free at follow-up (mean, 23 months) and postoperative cancer surveillance was not impaired by the combined procedures. The versatility of reduction mammaplasty allows this procedure to be performed in conjunction with partial mastectomy for any tumor location. Combining these procedures in patients with macromastia provides numerous therapeutic benefits at low cost, while reducing breast distortion and preserving symmetry. PMID- 11884820 TI - Rehabilitation of the replanted upper extremity. AB - A concise protocol for the rehabilitation of the upper extremity after replantation is presented with special emphasis in the early mobilization of the hand. PMID- 11884821 TI - Osteocutaneous posterior interosseous flap for reconstruction of the metacarpal bone and soft-tissue defects in the hand. AB - A vascularized bone segment of the ulna together with a posterior interosseous fasciocutaneous flap is harvested, including a cuff of the extensor pollicis longus muscle. The authors treated five male patients with metacarpal bone and soft-tissue defects of the hand using a distally based island osteocutaneous posterior interosseous flap. Their ages at the time of surgery ranged from 15 to 37 years (mean, 24 years). The bone defects were in the first metacarpal in three cases, the fourth metacarpal in one, and the fifth metacarpal in one. The length of the donated ulna ranged from 3 to 7 cm (mean, 5 cm). The follow-up period ranged from 5 to 92 months (mean, 39 months). All flaps survived completely. The posterior interosseous flap provides thin skin of good texture, together with vascularized bone, for a one-stage reconstruction of the metacarpal bone and soft tissue defects in the hand. PMID- 11884822 TI - Sensory reconstruction of the fingertip using the bilaterally innervated sensory cross-finger flap. AB - The authors present a series of 15 patients with large soft-tissue defects of the fingertips as a prospective, nonrandomized study. In all cases, reconstruction was achieved using a bilaterally innervated sensory cross-finger flap. This sensory fasciocutaneous flap relies on the dorsal branch of the proper digital nerves, which branch off at the level of the head of the proximal phalanx; sensory supply to the dorsal skin of the middle phalanx is thus ensured. The reconstructive procedure consists of two steps. First, the contralateral dorsal branch of the proper digital nerve is elevated with the flap at proximal interphalangeal joint level. Microsurgical coaptation is performed to the proximal nerve stump of the injured fingertip. After 3 weeks, when the pedicle is dissected, the second nerve is dissected and coapted. Clinical results were evaluated after 12 months. Because the regenerative distance is only 1.5 to 2.5 cm, good sensory regeneration should be expected. In nine of 16 flaps, sensory quality of S2+ (Highet) was present in the flap after 3 weeks. After 12 months, two-point discrimination was present in all patients, the values ranging between 2 and 6 mm (for two-point discrimination), with an average of 3.6 mm. The rate of complications was low. With acceptable additional operative action, a good functional result can be achieved. The indications of this method are discussed in comparison with other methods of fingertip reconstruction. PMID- 11884823 TI - A new surgical technique for the treatment of high common peroneal nerve palsy. AB - In this article, the authors introduce a new procedure for the treatment of high common peroneal nerve palsy. The principle of this technique consists of the neurotization of the anterior tibial nerve (deep peroneal nerve) with the bundle composed of the nerves to the soleus and lateral head of gastrocnemius muscles. The authors used this procedure for eight children who had permanent common peroneal nerve palsy caused by the injection of diclofenac in the gluteal region and for a 25-year-old male patient whose common peroneal nerve was transected near the gluteal region by a stab wound. For the cases in which paralysis was less than 8 months in duration, the results are satisfactory. PMID- 11884824 TI - Free-tissue transfer in patients with peripheral vascular disease: a 10-year experience. AB - Advances in free-tissue transfer have allowed for lower limb salvage in patients with significant peripheral vascular disease and limb-threatening soft-tissue wounds. The authors retrospectively reviewed their 10-year experience with free flaps for limb salvage in patients with peripheral vascular disease to assess postoperative complication rates and long-term functional outcome. They identified all patients undergoing free-tissue transfer with significant peripheral vascular disease and otherwise unreconstructible soft-tissue defects. Charts were reviewed for perioperative and long-term outcome. Parameters studied included perioperative morbidity and mortality, flap success, bypass graft patency, ambulatory results, and long-term limb and patient survival. Survival data were analyzed using life-table analysis, Kaplan-Meier survival analysis, and Cox testing. A total of 79 flaps were examined in 75 patients with peripheral vascular disease from July of 1990 to November of 1999. All patients would have required a major amputation had free-tissue transfer not been performed. Mean age was 60 years, average hospital stay was 32 days, and perioperative mortality was 5 percent. Within the first 30 days after operation, there were four cases of primary flap loss, and another two were lost as the result of bypass graft failure (8 percent); five of these cases resulted in amputation. There were no primary flap failures after 30 days. Follow-up ranged to 91 months (mean, 24 months). During this time, another 14 limbs were lost, most commonly because of progressive gangrene and/or infection in sites remote from the still-viable free flap. Using Kaplan-Meier survival analysis, 5-year flap survival was 77 percent, limb salvage 63 percent, and patient survival 67 percent. Sixty-six percent of patients were able to ambulate independently with the use of their reconstructed limb at least 1 year after hospital discharge, although some of these later went on to amputation. Free-tissue transfer for lower extremity reconstruction can be performed with acceptable morbidity and mortality in patients with peripheral vascular disease. Flap loss is low, and limb salvage, ambulation, and long-term survival rates in these patients are excellent. PMID- 11884825 TI - Versatility of the medial plantar flap: our clinical experience. AB - The medial plantar flap presents an ideal tissue reserve, particularly for the reconstruction of the plantar and palmar areas, which require a sensate and unique form of skin. In the past 5 years, the authors performed 16 free flaps, 10 locally pedicled flaps, and five cross-leg flaps on 31 patients for the reconstruction of palmar and plantar defects. All flaps transferred to the palmar area survived, providing good color match and sufficient bulkiness. The overall results were satisfactory in terms of function and sensation, and no complications related to flap survival in the plantar area were observed. All flaps used to cover defects in the heel and ankle region adapted well to their recipient areas, and all lower extremities remained functional. Because the medial plantar flap presents glabrous, sensate skin with proper bulkiness and permits the movement of underlying structures, the authors advocate its use and view this procedure as an excellent alternative in the reconstruction of palmar and plantar weight-bearing areas. PMID- 11884826 TI - Reconstruction of short lower leg stumps with the osteomusculocutaneous latissimus dorsi-rib flap. AB - To avoid a more proximal amputation at the distal part of the thigh, and when the knee joint is preserved, it is possible to lengthen short lower leg stumps. The authors report five cases in which the latissimus dorsi-rib flap was used to achieve a satisfactory functional prosthetic result. The bone segment is long enough to both lengthen the stump and allow its extremities to be firmly fixed to the tibia. Depending on the remaining tibia length, one or two ribs were included in the flap. The procedure allowed achievement of a 5-cm to 9-cm lengthening of the tibia. Bone healing time was 5 to 6 months and allowed prosthetic rehabilitation and ambulating 5 to 7 months after surgery. Final range of motion of the knee joint is compatible with normal ambulating, and the prosthesis is well tolerated. This procedure, which provides a large amount of skin, muscle, and bone, is very effective for reconstruction of short lower leg stumps. PMID- 11884827 TI - T-incision technique in distal hypospadias: a modification of meatal advancement and glanuloplasty. AB - Hypospadias is among the most common of the congenital anomalies. Distal hypospadias refers to an orifice in the distal third of the penile shaft. Correction of distal hypospadias requires different techniques, depending on the location of the meatus. Simple advancement techniques can be used for most distal hypospadias, whereas hypospadias with chordee requires reconstruction of a urethra. The meatoplasty and glanuloplasty procedures developed by Duckett have become standard operations to correct these lesions. Complications such as meatal stenosis, meatal retraction, and fish mouth-like meatus can be seen after meatal advancement and glanuloplasty ("MAGPI"), though it usually yields good results. In an attempt to avoid the complications associated with the meatal advancement and glanuloplasty procedure, the authors added a modification to the procedure for those hypospadias cases located in the coronal sulcus or its distal part. As an addition to conventional meatal advancement and glanuloplasty, a transverse incision on top of the vertical incision was made so that the urethra was supported by lateral triangle flaps created on the glans. Lateral triangle flaps of the glans were sutured to the dorsal aspect of the urethra advanced from the previous position. Thus, stress on the urethra was lessened and meatal retraction was prevented. When closure was performed with a T incision, an M-shaped, zigzag incision line was placed instead of a circular incision line. Therefore, stenosis was prevented and a vertical meatus with good cosmetic appearance was obtained. Fifty-three boys aged 4 to 7 years were operated on with this technique and were followed for 2.4 years. Good functional and cosmetic results were achieved in most of the cases. PMID- 11884828 TI - Long-term fate of the bony component in neophallus construction with free osteofasciocutaneous forearm or fibula flap in 18 female-to-male transsexuals. AB - Female-to-male transsexuals have been operated on in the authors' department since 1975. Between 1981 and 1995, 46 patients underwent neophallus construction with a free osteofasciocutaneous forearm or fibula flap. The bony part of these flaps is embedded in tissue with excellent blood circulation, has no contact with the skeleton, and is free of mechanical stress. To evaluate the long-term fate of the bony component of these flaps, the authors examined 18 of the 46 patients (39.1 percent) who had received a neophallus by means of one of these methods (12 with forearm and six with fibula flap) and who were willing to participate in the updating of the results of the previous two decades; this represented a follow-up of 5 to 112 months postoperatively (average, 27.4 months). The following investigations were undertaken: clinical and radiologic examination, bone scintigraphy, magnetic resonance imaging, and histologic examination of the neophallus bony component. In all patients, the clinical examination showed no significant variations in the shape and rigidity of the neophallus bone. The radiologic examination showed a compact bone structure, and the magnetic resonance imaging proved the vitality of the neophallus in all patients, with no significant changes over time. Bone scintigraphy did not prove to be useful in determining the long-term fate of the neophallic bony component. Histologically, subperiosteal neoformation of fibrous bone was shown, whereas the lamellar cortical bone was predominantly avital. The results of this study reveal the vitality of the bony component in neophallus construction with free osteofasciocutaneous flaps. Even 112 months after the procedure, it provided sufficient stiffness for sexual intercourse. This continuing adequate rigidity of the bony component, in addition to the well-known advantages of the free osteofasciocutaneous flap, is further evidence of its usefulness in neophallus construction. PMID- 11884830 TI - Rat extramedullary adipose tissue as a source of osteochondrogenic progenitor cells. AB - Human liposuction aspirates contain pluripotent adipose-derived mesodermal stem cells that have previously been shown to differentiate into various mesodermal cell types, including osteoblasts and chondrocytes. To develop an autologous research model of bone and cartilage tissue engineering, the authors sought to determine whether rat inguinal fat pads contain a similar population of osteochondrogenic precursor cells. It was hypothesized that the rat inguinal fat pad contains adipose-derived multipotential cells that resemble human adipose derived mesodermal stem cells in their osteochondrogenic capacity. To test this, the authors assessed the ability of cells isolated from the rat inguinal fat pad to differentiate into osteoblasts and chondrocytes by a variety of lineage specific histologic stains. Rat inguinal fat pads were isolated and processed from Sprague-Dawley rats into a fibroblast-like cell population. Cell cultures were placed in pro-osteogenic media containing dexamethasone, ascorbic acid, and beta-glycerol phosphate. Osteogenic differentiation was assessed at 2, 4, and 6 weeks. Alkaline phosphatase activity and von Kossa staining were performed to assess osteoblastic differentiation and the production of a calcified extracellular matrix. Cell cultures were also placed in prochondrogenic conditions and media supplemented with transforming growth factor-beta1, insulin, transferrin, and ascorbic acid. Chondrogenic differentiation was assessed at 2, 7, and 14 days by the presence of positive Alcian blue staining and type II collagen immunohistochemistry. Cells placed in osteogenic conditions changed in structure to a more cuboidal shape, formed bone nodules, stained positively for alkaline phosphatase activity, and secreted calcified extracellular matrix by 2 weeks. Cells placed in chondrogenic conditions formed cartilaginous nodules within 48 hours that stained positively for Alcian blue and type II collagen. The authors identified the rat inguinal fat pad as a source of osteochondrogenic precursors and developed a straightforward technique to isolate osteochondrogenic precursors from a small animal source. This relatively easily obtained source of osteochondrogenic cells from the rat may be useful for study of tissue engineering strategies and the basic science of stem cell biology. PMID- 11884832 TI - Prefabricated buccal mucosa-lined flap in an animal model that could be used for vaginal reconstruction. AB - Congenital vaginal aplasia, gynecological tumor excision, and male-to-female sex surgery are three clinical conditions in which the plastic surgeon is involved in vaginal reconstruction. Skin-lined or skin-grafted local flaps are currently used, but for many reasons, keratinized skin is not the ideal lining for such a moist cavity because it leads to dryness, desiccation, maceration of the skin, and even hair growth in the cavity. The purpose of this study was to create a subcutaneous cavity lined with mucosa in an area with a predictable blood supply. The abdominal area supplied by the deep circumflex iliac vessels was chosen. Six minipigs were used. Strips of tongue buccal mucosa formed the lining; if additional tissue was required, it was taken from the mucosal aspect of the cheek. The mucosa was expanded by using multiple stab incisions. The mucosa was sutured onto the fascia supplied by the deep circumflex iliac vessels, and the skin incision was closed over a silicone sheet to prevent adhesion to the underlying mucosa. This was left for 1 week to allow the mucosa to take. The prefabricated fascial flap was rolled over a silicone stent and was closed longitudinally to form a cylindrical shape. The flap was placed in a subcutaneous pocket in the right inguinal area. The caudal end was left open and was sutured to the surrounding skin. The silicone stent was used to keep the cavity patent and to prevent adhesions in the early stage of the healing process. Regular digital examination was performed to assess patency and contour; endoscopy allowed assessment of mucosa viability. This method of producing a mucosa-lined flap may provide a solution to the difficult problem of vaginal reconstruction. PMID- 11884834 TI - The relative importance of the deep and superficial vascular systems for delay of the transverse rectus abdominis musculocutaneous flap as demonstrated in a rat model. AB - The use of some form of delay maneuver for "high-risk" patients before transfer of the superior pedicled lower transverse rectus abdominis musculocutaneous (TRAM) flap for breast reconstruction has augmented the rate of success in both the experimental and clinical arenas. A common method of vascular delay has been the bilateral division of both the superficial inferior epigastric and deep inferior epigastric vessels. Whether all of these must be divided to adequately effect the delay is unknown. For that matter, the relative importance of the superficial versus the deep vascular systems is unclear. To investigate this uncertainty, a delay was attempted in 61 Sprague-Dawley rats by division of either the superficial inferior epigastric or deep cranial epigastric vessels (the latter is the homologue to the human deep inferior epigastric) in unilateral or bilateral fashion. Division of the contralateral superficial inferior epigastric vessel resulted in significantly greater TRAM flap survival than either ipsilateral or bilateral superficial inferior epigastric vessel division (p = 0.0034 or p = 0.0093, respectively). Division of the ipsilateral or bilateral deep cranial epigastric vessel resulted in significantly greater flap survival than just contralateral deep cranial epigastric vessel division (p = 0.0034 or p = 0.006, respectively). No significant difference was observed between the group having contralateral superficial inferior epigastric or groups with ipsilateral deep cranial epigastric division, implying that either alone would be efficacious to achieve the desired delay effect. This would allow the other vascular system to be retained intact for later potential salvage maneuvers as needed. PMID- 11884836 TI - Reconstruction of an extensive hemipelvectomy defect using a pedicled upper and lower leg in-continuity fillet flap. PMID- 11884837 TI - Toe-finger switch operation. PMID- 11884838 TI - On-top plasty for duplicated hallux with hallux varus using microsurgical transfer of the whole toe. PMID- 11884839 TI - Transport distraction osteogenesis: a new method to heal adult calvarial defects. AB - Popularized by Gavril Ilizarov in the 1960s, monofocal distraction osteogenesis has become a well-established method of endogenous bone engineering. This revolutionary surgical technique has significantly augmented the available reconstructive orthopedic and craniomaxillofacial procedures. Bifocal distraction osteogenesis, or bone transportation, is a modification of monofocal distraction that involves moving a free segment of living bone to fill an intercalary bone defect. Bifocal distraction has been applied successfully to reconstruct complex mandibular and long bone defects. Because traumatic or postsurgical calvarial defects do not spontaneously heal in humans older than 18 to 24 months of age, we hypothesized that bifocal distraction osteogenesis could be applied to the skull to close critical size calvarial defects. Critical size (15 x 15 mm) calvarial defects were created in eight New Zealand White rabbits. Next, a 15-mm x 10-mm calvarial box osteotomy was created just anterior to the skull defect. This osteotomy created a free bone segment that could be transported. A custom-made transport distraction device was fixed into place and the skin incision was closed. After a 4-day latency period, the distraction device was activated (0.5 mm once daily for 30 days) in seven animals; the distraction device in one animal was not activated and served as a control. All animals underwent 30 days of consolidation and were then killed. Radiographs and computed tomographic scans were performed at the following time points: end of latency period (postoperative day 4), mid-distraction (postoperative day 19), and end of consolidation period (postoperative day 64). Gross and histologic analysis was performed to evaluate the quality of the bony regenerate. The control animal healed with a fibrous union. Complete closure of the skull defects was observed in five of seven rabbits at the end of the consolidation period. One animal was removed from the study because of an early loosening of the distraction device, and one was removed because of device failure. Of the remaining five animals that completed the distraction protocol, radiographs and computerized tomographic scans showed successful ossification in all five rabbits at the end of the consolidation period. This study suggests that transport distraction osteogenesis is a promising technique that may be applied to a variety of commonly encountered craniofacial problems such as nonhealing calvarial defects. PMID- 11884840 TI - Isolated zygomatic arch fracture: report on a modified surgical technique. PMID- 11884841 TI - Retroauricular prefabricated chondrofasciocutaneous flap for reconstruction of the columella. PMID- 11884842 TI - Bilateral triangular flaps for the correction of posttracheostomy scars: a simpler approach. PMID- 11884843 TI - Refinements in reduction mammaplasties from a solely inframammary approach. PMID- 11884845 TI - Scar assessment tools: implications for current research. AB - Scarring is considered a major medical problem that leads to cosmetic and functional sequelae. Scar tissue is clinically distinguished from normal skin by an aberrant color, rough surface texture, increased thickness (hypertrophy), occurrence of contraction, and firmness. Marked histologic differences are the change in dermal architecture and the presence of cells such as the myofibroblast. Many assessment tools are available for analysis of pathologic conditions of the skin; however, there is no general agreement as to the most appropriate tools for evaluation of scar tissue. This review critically discusses currently available objective measurement tools, subjective assessment tools, and potential devices that may be available in the future for scar assessment. PMID- 11884846 TI - Long-term clinical experience with the Ruiz-Cohen intraoperative arterial expanders. PMID- 11884847 TI - Understanding the nasal airway: principles and practice. AB - LEARNING OBJECTIVES: After studying this article, the reader should be able to: 1. Describe the soft-tissue, cartilaginous, and bony anatomy of the nose. 2. Describe the anatomy and function of the nasal valves. 3. Discuss the governing physiologic principles responsible for airflow dynamics. 4. Discuss the various functions of the nose. 5. Demonstrate an appropriate evaluation of the nasal airway. 6. Discuss the differential diagnosis of nasal obstruction. 7. Discuss appropriate management options for nasal airway obstruction. The nose is a complex, multifunctional organ that requires respect and understanding from the rhinoplasty surgeon. The etiologic and pathologic characteristics of each patient's nasal airway problem determine the treatment of the nasal airway. Frequently, medical management is sufficient without operative intervention. Recent advances have shown that nasal valves in airway patency may play a more important role than the septum. The rhinoplasty surgeon's understanding of the anatomy and physiology of the nasal airway, along with the causes of obstruction, can pave the way for a proper evaluation and appropriate management of nasal airway problems. Lack of understanding can result in misdiagnosis and mismanagement. This article outlines current concepts of medical and surgical management of nasal airway problems and discusses in detail the key concepts and principles in the practical management of the nasal airway. PMID- 11884848 TI - Lasers in office-based settings: establishing guidelines for proper usage. PMID- 11884849 TI - The superficial lateral canthal tendon: anatomic study and clinical application to lateral canthopexy. AB - Most patients who undergo facial cosmetic surgery procedures that could cause lower eyelid retraction or ectropion should have an additional surgical procedure to support the lower eyelid and lateral canthus. The lower eyelid should be supported when performing laser planing of the eyelid; midface elevation through a lower eyelid incision approach; or conventional blepharoplasty, in patients with lower eyelid laxity. Suspending the lateral canthus by surgically altering the lateral canthal tendon is a proven technique that can provide support for the lower eyelid. However, a technique of this complexity may be unnecessary for most cosmetic surgery patients. To increase understanding of the fascial support system of the lateral canthus, four fresh cadaver dissections were performed to investigate the attachments of the lateral canthus to the lateral orbital rim. The most commonly appreciated attachment between the eyelids and the lateral orbital rim is the lateral canthal tendon (the lateral canthal raphe). However, the lateral canthus also is attached to the orbital rim at a more superficial level through the septum orbitale. This superficial fascial plane may be modified and used as a structure to stabilize or suspend the lateral canthus. This structure is defined in this article as the "superficial lateral canthal tendon." PMID- 11884852 TI - Correction of lateral brow ptosis: a nonendoscopic subgaleal approach. AB - The authors present their experience with a relatively uncomplicated, rapid technique for elevation of the lateral eyebrow and a simultaneous correction of eyelid hooding that is secondary to the descent of the eyebrow. The procedure is designed for all patients requiring lateral brow elevation, either separately or in combination with other procedures. The authors describe and illustrate their technique. PMID- 11884854 TI - New technique of plication for miniabdominoplasty. AB - The authors present a new method of plication of abdominal fascia performed in 42 patients who underwent surgery for miniabdominoplasty between September of 1998 and February of 2000. The design consisted of a horizontal half-moon on the infraumbilical fascia with high lateral tension, similar to the one that is performed on the skin, achieving an improvement in the muscle-aponeurotic tension of the whole abdomen without requiring a supraumbilical dissection or undermining. All patients (n = 42) had a superficial and a deep conventional liposuction of the abdomen, flanks, and posterior trunk with the wet technique. The rate of minor complications was 59.5 percent. Twenty patients had seromas, three patients had dog-ears and one had cutaneous ischemia (epidermolysis). There were no cases of major complications such as tissue necrosis, infections, deep venous thrombosis, pulmonary emboli, or fat embolus syndrome. Patients received follow-up examination between 6 months and 2 years after surgery (average, 15 months). The results were excellent, and the patients were completely satisfied. PMID- 11884857 TI - And for my next trick...! PMID- 11884856 TI - Are plastic surgery advertisements conforming to the ethical codes of the american society of plastic surgeons? AB - Cosmetic surgeons have increasingly come under fire for using advertisements that may be deceptive or intended for the solicitation of vulnerable consumers. However, aesthetic surgery is a growing business that relies heavily on advertising to survive. To prevent the use of deceptive advertisements, the American Society of Plastic Surgeons has developed a code of ethics for its physician members. We conducted a study to determine the prevalence of cosmetic surgery advertisements considered objectionable by the lay public. These advertisements were published in the Yellow Pages of the 10 largest U.S. cities. Because all of the advertisements in this study contained the American Society of Plastic Surgeons logo, we also determined whether its members are upholding the ethical code of advertising. We asked a convenience sample of 50 participants to rate 104 advertisements using four yes/no questions derived from the code of ethics and one overall yes/no question regarding whether the advertisement was objectionable. We obtained the mean percentage of "yes" responses for each advertisement, from the total sample, for each question. We found that the study participants felt that 25 percent of the advertisements used images of persons or facsimiles that falsely and deceptively created unjustified expectations of favorable results. The participants responded that 22 percent of the advertisements appealed primarily to the layperson's fears, anxieties, or emotional vulnerabilities. In addition, 18 percent of the advertisements were considered to be objectionable. Discretion is currently left up to physicians as to the ethical nature of their advertisements. Although the majority of American Society of Plastic Surgeons members uphold the ethical code of advertising, there are still a substantial number of published advertisements that the average consumer considers to be in violation of this code. PMID- 11884858 TI - There is no accounting for accountability. PMID- 11884859 TI - Botulinum toxin. PMID- 11884860 TI - Gigantic methameric seborrheic keratosis. PMID- 11884861 TI - Radiation therapy in postmastectomy TRAM reconstruction. PMID- 11884862 TI - Pubic hair reconstruction using minigrafts and micrografts. PMID- 11884863 TI - The responsibility of plastic surgeons to help patients stop smoking. PMID- 11884864 TI - Successful primary microsurgical replantation of an avulsed penis. PMID- 11884865 TI - Medicare reform house floor speech by Congressman Dr. Gregory Ganske, June 6, 2001. PMID- 11884866 TI - Breast reconstruction with the internal mammary artery pedicled fasciocutaneous island flap: description of a new flap. PMID- 11884867 TI - The use of clonidine in facial plastic surgery. PMID- 11884868 TI - Successful free flap transfer to the lower extremity in a patient with raynaud syndrome of the four extremities. PMID- 11884870 TI - Is pharmacologic vitreolysis brewing? PMID- 11884871 TI - To laser the ridge or not laser the ridge, that is the question. PMID- 11884872 TI - Guidelines for using verteporfin (visudyne) in photodynamic therapy to treat choroidal neovascularization due to age-related macular degeneration and other causes. AB - OBJECTIVE: Guidelines were developed based on best available scientific data as well as consensus of expert opinion in absence of controlled clinical trial data to: 1) assist ophthalmologists with selection of patients for whom photodynamic therapy with verteporfin, termed "verteporfin therapy," should be considered; and 2) offer suggestions regarding treatment, follow-up, and re-treatment. METHODS: Consensus from roundtable of retina specialists who either participated in randomized clinical trials evaluating verteporfin therapy or had clinical experience with verteporfin therapy was based on results of these trials and expert opinion. Additional input and advice were received from representatives on behalf of the Macula Society, the Retina Society, and the Vitreous Society, as well as principal investigators of randomized clinical trials evaluating verteporfin therapy. RESULTS: Patient selection criteria included the following: 1) in cases due to age-related macular degeneration (AMD), lesion composition either predominantly classic choroidal neovascularization (CNV) or occult with no classic CNV; 2) CNV location subfoveal or so close to the foveal center that conventional laser photocoagulation treatment almost certainly would extend under the center; 3) lesion etiology from AMD, pathologic myopia, or other causes in which the outcome without treatment is likely to be worse than with treatment; 4) vision at a level where further loss would be recognized as detrimental to the quality of life of the patient. Criteria did not include lesion size, except in cases composed of occult with no classic CNV in AMD in which therapy for lesions >4 Macular Photocoagulation Study (MPS) disc areas usually should be considered only when presenting with lower levels of best-corrected visual acuity. Criteria also did not include patient age, history of systemic arterial hypertension, or prior laser photocoagulation. Therapy should occur ideally within 1 week of the initial fluorescein angiogram on which the clinical decision to treat is based. Patients should return for follow-up at least as often as every 3 months after any initial or subsequent treatment to determine if there is fluorescein leakage from CNV. Re-treatment should be considered as often as every 3 months if fluorescein leakage from CNV is noted at that time. Re-treatment could be deferred if the biomicroscopic and fluorescein angiographic appearance of the lesion is unchanged and shows minimal leakage, especially when there is no subretinal fluid or fluorescein leakage from CNV underlying the center of the foveal avascular zone. Patients should avoid exposure of skin or eyes to direct sunlight or bright indoor light for 48 hours after treatment or until resolution of any swelling or discoloration from extravasation. CONCLUSION: These recommendations provide guidelines on the role of verteporfin therapy in the management of CNV due to AMD and other causes. Revisions of these guidelines may be required as new data become available. PMID- 11884873 TI - Factors associated with reduced visual acuity during long-term follow-up of patients with idiopathic central serous chorioretinopathy. AB - PURPOSE: To investigate factors associated with reduced visual acuity during long term follow-up of patients with idiopathic central serous chorioretinopathy (ICSC). METHODS: Retrospective consecutive case series that included patients with ICSC who were younger than 50 years of age at the time of initial examination and were followed up for > or =3 years. RESULTS: The mean follow-up for 101 involved eyes of 61 patients was 9.8 years (median, 8.0 years). Eyes were stratified into two groups based on visual acuity at the final examination: Group 1, visual acuity of 2040 or better; and Group 2, visual acuity of worse than 2040. Findings identified as potential risk factors for reduced vision at the final follow-up examinations for Group 1 versus Group 2 included the following: macular retinal pigment epithelium atrophy (90.8% versus 96.0%, respectively; P = 0.68); persistent pigment epithelial detachment or persistent subretinal fluid (5.3% versus 28.0%, respectively; P = 0.004); recurrences (39.5% versus 68.0%, respectively; P = 0.020); laser treatment (28.9% versus 32.0%, respectively; P = 0.80); and submacular choroidal neovascularization (0.0 versus 8.0%, respectively; P = 0.059). CONCLUSIONS: Factors associated with reduced visual acuity during long-term follow-up of patients with ICSC included persistent pigment epithelial detachment and/or subretinal fluid, recurrences, and submacular choroidal neovascularization. PMID- 11884874 TI - A long-term follow-up study of severe variant of central serous chorioretinopathy. AB - PURPOSE: To facilitate understanding of the long-term course and visual outcome of a severe variant of central serous chorioretinopathy. DESIGN: Consecutive observational case series. PATIENTS AND METHODS: The authors reviewed 25 patients with multifocal posterior pigment epitheliopathy and bullous retinal detachment, who had a mean follow-up time of 10.6 years (range, 6-22 years), with reference to the demographic feature, fundus changes, recurrence, and final anatomic and visual outcome. Two patients underwent optical coherence tomography. RESULTS: The patients were 21 men and 4 women, with a mean age at disease onset of 43.1 years (range, 30-63 years). Twenty-one patients were otherwise healthy, and four developed ocular disease during systemic corticosteroid therapy for metabolic or autoimmune diseases including systemic lupus erythematosus. The disease was bilateral in 21 patients (84%). Nine patients (36%) presented initially with classic central serous chorioretinopathy, followed by its severe variant 7 months to 9 years later. Active disease was characterized by multifocal exudative lesions in the posterior pole and bullous retinal detachment with shifting subretinal fluid in the inferior periphery. Optical coherence tomography of exudative lesions disclosed cloudy and fibrinous subretinal fluid. The exudative lesions were self-limited or responded to photocoagulation. During the follow-up period, 13 patients (52%) showed 1 to 5 recurrent disease, but the disease eventually became quiescent with multifocal atrophic scars in the posterior pole with or without atrophic tracts in the inferior periphery. Final best-corrected visual acuity was 2020 or better in 24 of 46 affected eyes (52%) of 25 patients and 2040 or better in 37 eyes (80.4%). CONCLUSIONS: A severe variant of central serous chorioretinopathy characterized by multifocal posterior exudations and bullous inferior retinal detachment with shifting subretinal fluid may affect otherwise healthy, middle-aged males or individuals receiving systemic corticosteroid therapy for metabolic or autoimmune diseases. Exudative chorioretinal lesions are self-limited or respond to photocoagulation. Recurrence is common, but the disease eventually becomes quiescent with favorable visual acuity unless the macula is damaged. PMID- 11884875 TI - "On" response dysfunction in multifocal posterior pigment epitheliopathy. AB - PURPOSE: To determine the electroretinographic properties of one patient with multifocal posterior pigment epitheliopathy. METHODS: Rod and cone electroretinograms (ERGs) and photopic ERGs elicited by long-duration stimuli were studied in a patient with multifocal posterior pigment epitheliopathy. RESULTS: The amplitudes of both the rod and cone ERGs were significantly reduced. Photopic ERGs elicited by long-duration stimuli demonstrated that the b-wave ("on" response) was abolished but the d-wave ("off" response) was reduced by only amplitude. CONCLUSION: The ERG findings in multifocal posterior pigment epitheliopathy indicate that there is dysfunction not only of the photoreceptors but also in the signal transmission specific for the "on" pathway. PMID- 11884876 TI - Fluorescein angiographic findings in primary intraocular lymphoma. AB - PURPOSE: Primary intraocular lymphoma (PIOL), also known as primary central nervous system lymphoma, is a rare yet blinding and fatal disease. Often presenting with ocular involvement, it can masquerade as posterior or intermediate uveitis, thus delaying diagnosis. A noninvasive ancillary test such as fluorescein angiography could be helpful in raising the level of suspicion in the diagnosis of this disease. METHODS: Results of fluorescein angiography (FA) and clinical characteristics of 17 patients (31 eyes) who presented to the National Eye Institute with the diagnosis of PIOL (confirmed by histopathologic analysis) were reviewed. RESULTS: The most common angiographic characteristics included disturbances at the level of the retinal pigment epithelium (RPE), such as granularity (19 eyes [61%]), blockage (17 eyes [55%]), and late staining (14 eyes [45%]). These changes are well correlated to histopathologic findings of lymphoma cells located between the RPE and Bruchs membrane. Perivascular staining or leakage and cystoid macular edema were rare. Other less common findings included pigment epithelial detachments and punctate hyperfluorescent lesions. Clinical characteristics found in eyes for which results of FA were available included vitreitis (29 eyes [94%]), subretinal infiltrates (19 eyes [61%]), and anterior chamber cells (10 eyes [32%]). In some cases, clinical examination did not correlate with FA findings. CONCLUSIONS: Although PIOL may present with a normal angiographic phenotype, extensive RPE changes demonstrated by FA, combined with the absence of perivascular staining or leakage and macular edema, may be associated with and distinctive of PIOL. PMID- 11884877 TI - Interpretation of flow cytometric measurement of lymphocytes after fluorescein angiography. AB - PURPOSE: By use of flow cytometry (FCM), lymphocyte subsets were evaluated with fluorochrome-labeled monoclonal antibodies (MoAbs). Recent fluorescein angiography (FA) produces temporary elevation of serum background fluorescence at certain wavelengths of light, producing falsely decreased lymphocyte subset quantitations. The authors evaluated the duration of this effect on one subset of lymphocytes after FA. METHODS: CD4 counts were determined by FCM before and 10 minutes, 1, 6, and 24 hours after injection of fluorescein dye in 12 patients. The MoAbs used were directly conjugated to fluorescein isothiocyanate (FITC) or phycoerythrin (PE). RESULTS: Using FITC-labeled MoAbs, falsely decreased CD4 counts occurred in all patients at 10 minutes and in all but one patient 1 hour after injection. Return to baseline levels occurred in 50% (95% confidence interval [CI], 0.21, 0.79) by 6 hours and in 75% (95% CI, 0.43, 0.95] by 24 hours. No such effect was observed using PE-labeled MoAbs. CONCLUSIONS: Falsely decreased CD4 values as determined by FCM were present immediately after FA in all 12 patients and persisted 24 hours in some patients when FITC-labeled MoAbs were used. CD4 evaluation should be delayed in patients who have undergone recent FA or the analysis should be performed with PE-labeled MoAbs. PMID- 11884878 TI - Diode laser photocoagulation to the ridge and avascular retina in threshold retinopathy of prematurity. AB - PURPOSE: To report results applying diode laser photocoagulation to both the peripheral avascular retina and the ridge in stage 3+ threshold retinopathy of prematurity. METHODS: The authors performed a retrospective review of 82 consecutive eyes in 43 preterm infants with threshold disease who had both the peripheral avascular retina and the ridge treated with diode laser photocoagulation. With a minimum follow-up of 3 months, these eyes were evaluated for intraoperative and postoperative complications and long-term anatomic results. RESULTS: A favorable anatomic outcome occurred in 79 eyes (96%). There were no intraoperative complications. Postoperative intraocular hemorrhage occurred in eight eyes (10%) and resolved without sequelae. Supplemental laser was required in only two eyes (2%). CONCLUSIONS: Diode laser photocoagulation to the ridge and peripheral avascular retina in threshold retinopathy of prematurity is associated with a favorable anatomic outcome. The risk of postoperative intraocular hemorrhage and the need for supplemental laser photocoagulation is low. PMID- 11884879 TI - Ciliary detachment after pars plana vitrectomy: an ultrasound biomicroscopic study. AB - PURPOSE: To determine the incidence, duration, risk factors, and clinical outcomes for ciliary detachment after pars plana vitrectomy (PPV). METHODS: A total of 109 eyes of 103 patients who underwent PPV for various disease entities were included. Ultrasound biomicroscopy was applied to determine the tomographic features of the ciliary body before and 1, 3, and 7 days after the surgery. All eyes were then examined once weekly for 2 months. Demographic, preoperative, intraoperative, and postoperative parameters were evaluated to assess their predictive value in the formation of postvitrectomy ciliary detachment. RESULTS: Ciliary detachment was observed in 46 eyes (42%) after surgery and persisted for less than 3 weeks in 40 of 46 eyes. It most frequently occurred in eyes of patients with proliferative diabetic retinopathy (PDR) (64%) or retinal vascular obstructive diseases (RVO) (47%). Extensive retinal photocoagulation and retinal cryopexy positively predisposed to its formation whereas fluid-gas exchange had a protective effect. No clinical complications were observed in eyes with postoperative ciliary detachment. CONCLUSION: Ciliary detachment occurred frequently after PPV. A diagnosis of PDR or RVO and surgical procedures with extensive retinal photocoagulation and retinal cryopexy may have a higher incidence of its occurrence. PMID- 11884880 TI - Comparison of nonmydriatic digitized video fundus images with standard 35-mm slides to screen for and identify specific lesions of age-related macular degeneration. AB - PURPOSE: The purpose of this study was to compare nonmydriatic digitized images obtained using a digital imaging system (resolution of 640 x 480 pixels) with 35 mm slide images for detecting specific findings of age-related macular degeneration (AMD) and to evaluate its usefulness as a screening tool in detecting signs of AMD. METHODS: Seventeen consecutive patients (33 eyes) underwent digital color imaging (with a nonmydriatic, 45-degree, fundus camera attached to a digital back) and standard 35-mm, 30-degree retinal color photography of the fundus: posterior pole, nasal retina, and temporal retina. The images were later reviewed for the presence or absence of specific retinal findings. The images were not compressed. Primary outcome measures included the presence or absence of drusen, hard exudate, choroidal neovascularization (CNV), subretinal hemorrhage, retinal pigment epithelial (RPE) changes, subretinal fibrosis, pigment epithelial detachment (PED), and subretinal fluid. Presence of drusen, with or without any one of the other findings, and presence of disciform scar or geographic atrophy were positive indications in screening for AMD. RESULTS: Agreement between image type was highest for PED (97%), CNV (91%), and subretinal fibrosis (91%); and lowest for RPE changes (63%). Sensitivity, specificity, positive predictive value, and negative predictive values were determined using the 35 mm-slide images as the reference for comparison. Sensitivity ranged from 40% (hard exudates) to 75% (subretinal hemorrhage). Specificity ranged from 88% (drusen) to 100% (hard exudate, CNV, RPE changes, PED). Positive predictive value ranged from 67% (subretinal hemorrhage) to 100% (hard exudate, CNV, RPE changes, PED). Negative predictive value ranged from 44% (drusen) to 97% (PED). For the purposes of screening for any evidence of AMD, the system was 70% sensitive. CONCLUSION: This digital fundus imaging system with 640 x 480 pixel resolution has low sensitivity and high specificity, as compared with 35-mm slide images, for detection of early AMD, but higher sensitivity for late findings (CNV, scar, atrophy) of AMD. Because of sensitivity for detecting any AMD coupled with the low sensitivity for detecting CNV, the system is not useful for evaluating AMD patients who require close follow-up and who are at risk for more severe visual loss. PMID- 11884881 TI - Localization of rose bengal, aluminum phthalocyanine tetrasulfonate, and chlorin e6 in the rabbit eye. AB - PURPOSE: The localization and site of action of photosensitizers in the eye may be important for photodynamic therapy for fundus disorders but remain poorly understood for most agents. We investigated the intraocular localization of xanthene, phthalocyanine, and chlorin photosensitizers by using fluorescence microscopy and digital fundus fluorescence angiography. METHODS: Rose bengal (40 mg/kg), aluminum phthalocyanine tetrasulfonate (CASPc) (5 mg/kg), or chlorin e6 (2 mg/kg) was intravenously administered to albino rabbits. The eyes were enucleated and examined by means of fluorescence microscopy 5, 20, 60, and 120 minutes and 24 hours after dye injection. In vivo digital fundus fluorescence angiography with use of rose bengal (2-4 mg/kg), CASPc (2 mg/kg), and chlorin e6 (2 mg/kg) was performed. RESULTS: For all agents studied pathologically, there was moderate fluorescence from the choroid and retinal pigment epithelium 5 minutes after dye injection. Mild fluorescence detected from the photoreceptor outer segments at 5 minutes was increased at 20 minutes. Angiographic studies with use of rose bengal, CASPc, and chlorin e6 revealed differences in the pattern and rate of photosensitizer accumulation. CONCLUSIONS: Rose bengal, CASPc, and chlorin e6 accumulate rapidly in the choroid and retinal pigment epithelium and less rapidly in the outer retina. Differences in ocular localization of these photosensitizers were demonstrated. The significance of these findings for potential photodynamic therapy with these agents requires further investigation. PMID- 11884883 TI - Review: coats disease: the 2001 LuEsther T. Mertz lecture. PMID- 11884882 TI - Systemic non-Hodgkin's lymphoma with optic nerve infiltration in a patient with AIDS. AB - PURPOSE: To report the clinicopathologic features of a patient with AIDS and clinically regressed systemic non-Hodgkin's lymphoma who subsequently developed lymphomatous infiltration of the optic nerve and occlusion of the central retinal vein in both eyes. METHODS: The eyes of this patient were examined ophthalmologically and by fluorescein angiography. The eyes, brain, and body were obtained after death and studied by light microscopy. RESULTS: Ophthalmic examination and fluorescein angiography revealed optic nerve swelling and central retinal vein occlusion first in the left eye and shortly thereafter in the right eye. Postmortem histopathologic examination showed dense infiltration of both optic nerves by lymphoma as well as necrotizing vasculitis involving the retinal vessels near the left optic nerve head. Examination of the brain revealed lymphomatous involvement in the paraventricular region with associated necrosis and inflammation. No bone marrow recurrence or other residual systemic lymphoma was present. CONCLUSION: This case demonstrates that infiltrative optic neuropathy may occur as the sole ocular manifestation of disease recurrence in a patient with systemic non-Hodgkin's lymphoma otherwise thought to be in clinical remission. PMID- 11884884 TI - Diagnostic and therapeutic challenges. Consultants suspect either coagulopathy, hypertensive episode of unknown etiology or anticoagulation. PMID- 11884885 TI - Combined hamartoma of the retina and retinal pigment epithelium: optical coherence tomography. PMID- 11884886 TI - Visual improvement after pars plana vitrectomy and membrane peeling for vitreoretinal traction associated with combined hamartoma of the retina and retinal pigment epithelium. PMID- 11884887 TI - Indocyanine green can distinguish posterior vitreous cortex from internal limiting membrane during vitrectomy with removal of epiretinal membrane. PMID- 11884888 TI - Negative indocyanine green staining of epiretinal membranes. PMID- 11884889 TI - Scanning laser opthalmoscope findings in acute macular neuroretinopathy. PMID- 11884890 TI - Surgery for choroidal neovascularization in sympathetic opthalmia. PMID- 11884891 TI - Photodynamic therapy for choroidal neovascularization in fundus flavimaculatus. PMID- 11884892 TI - Macular hole formation following rupture of retinal arterial macroaneurysm. PMID- 11884893 TI - Postsurgical alterations in visual acuity, retinal vasculature, and retinal circulation times in Takayasu's disease. PMID- 11884894 TI - Retinal toxicity from accidental intraocular injection of depo-medrol. PMID- 11884895 TI - Polymorphisms 1704G/T, 2184A/G, and 2245G/A in the rage gene are not associated with diabetic retinopathy in NIDDM: pilot study. PMID- 11884896 TI - Single-bite suture for securing the infusion cannula in vitrectomy. PMID- 11884897 TI - Microsporidial endophthalmitis in a patient with acute myelogenous leukemia. PMID- 11884898 TI - Choroidal melanoma in a black patient with oculodermal melanocytosis. PMID- 11884899 TI - Macular buckling for retinal detachment due to macular hole in highly myopic eyes with posterior staphyloma. PMID- 11884900 TI - A new three-port cannular system for closed pars plana vitrectomy. PMID- 11884901 TI - Posterior retinal vasculitis associated with multiple sclerosis. PMID- 11884902 TI - A randomized controlled trial of two strategies to implement active sick leave for patients with low back pain. AB - STUDY DESIGN: Cluster randomized controlled trial. OBJECTIVE: To evaluate the effectiveness of two strategies to improve the use of active sick leave (ASL) for patients with low back pain. SUMMARY OF BACKGROUND DATA: ASL is a public sickness benefit scheme offered to promote early return to modified work for temporarily disabled workers. It was poorly used, and the authors designed two community interventions to strengthen the implementation of ASL based on the results of a study of barriers to use among back pain patients, employers, general practitioners (GPs), and local National Insurance Administration staff. METHODS: Sixty-five municipalities in three counties in Norway, randomly assigned to a passive intervention, a proactive intervention, or a control group. The interventions were targeted at patients on sick leave for low back pain for more than 16 days (n = 6176), their GPs, employers, and local insurance officers. The passive intervention included reminders about ASL on the sick leave form that GPs must complete, a standard agreement to facilitate ASL, targeted information, and a desktop summary for GPs of clinical practice guidelines for low back pain, emphasizing the importance of advice to stay active. The proactive intervention included these elements plus a resource person to facilitate the use of ASL and a continuing education workshop for GPs. The main outcome measure reported here is the proportion of eligible patients that used ASL. RESULTS: ASL was used significantly more in the proactive intervention municipalities (17.7%) compared with the passive intervention and control municipalities (11.5%, P = 0.018). CONCLUSIONS: A passive intervention that addressed identified barriers to the use of ASL did not increase its use. Although modest, a proactive intervention did increase its use. The main impact of the intervention was through direct contact and motivating telephone calls to patients. To the extent that GPs' practice was changed, it was either patient mediated or by patients bypassing their GP. PMID- 11884903 TI - Histologic evaluation of the efficacy of rhBMP-2 compared with autograft bone in sheep spinal anterior interbody fusion. AB - STUDY DESIGN: The sheep anterior lumbar spinal fusion model was used to study the efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2)-collagen composite in comparison with autograft to enhance spinal interbody fusion. Comparisons were drawn from temporal radiographic and end-point biomechanical and histologic data. OBJECTIVE: To analyze histologically the ability of rhBMP-2 to achieve complete arthrodesis between vertebral bodies. SUMMARY OF BACKGROUND DATA: Studies using rhBMP for enhancement of anterior interbody fusion have used numerous endpoints. However, systematic histologic evaluation of the fusion has not been conducted. METHODS: Twelve sheep underwent single-level anterior lumbar interbody fusion performed with a cylindrical fenestrated titanium interbody fusion device (INTER FIX, Medtronic Sofamor Danek, Inc., Memphis, TN). The device was filled either with rhBMP-2-collagen (n = 6) or autogenous iliac crest bone graft (n = 6). Radiologic evaluation was carried out at 2-month intervals, and all sheep were killed 6 months after surgery. Nondestructive biomechanical testing for stiffness to flexion, extension, and lateral bending moments, un decalcified histology, and qualitative and quantitative histologic evaluation were performed. RESULTS: Radiographs revealed a bony bridge anterior to the cage in five of six rhBMP-2-treated animals, whereas it was present only in one of five in the autogenous bone graft group. Segments treated with rhBMP-2 were 20% stiffer in flexion than autograft-treated segments at 6 months. Six of six in the rhBMP-2 group and two of six in the autograft group showed complete fusion. There was a significantly higher rate of bony continuity observed at the fenestrations of the rhBMP-2 group. Three times more number of cage fenestrations in the rhBMP 2 group demonstrated "all-bone" when compared with the autograft group (P < 0.001). Further, the scar tissue in and around the autograft-treated cages was 16 fold more (P < 0.01) than that seen for rhBMP-2-treated cages. CONCLUSIONS: The study demonstrates that rhBMP-2 can lead to earlier radiologic fusion and a more consistent increased stiffness of the segments when compared with autograft in sheep anterior lumbar interbody fusion. Furthermore, a three times higher histologic fusion rate is attainable with significantly reduced fibrous tissue around the implant when rhBMP-2 is used. PMID- 11884904 TI - Effect of chondroitinase ABC on matrix metalloproteinases and inflammatory mediators produced by intervertebral disc of rabbit in vitro. AB - STUDY DESIGN: Lumbar intervertebral discs in rabbit were cultured in the presence of chondroitinase ABC. The matrix metalloproteinases (MMPs) and inflammatory mediators produced in culture media were then analyzed. OBJECTIVES: To investigate the effect of chondroitinase ABC on MMPs and inflammatory mediators produced by intervertebral disc of rabbit in vitro. SUMMARY OF BACKGROUND DATA: The chemonucleolytic effect of chondroitinase ABC is caused by the decrease in the chondroitin sulfate, hyaluronan, and protein content of the nucleus pulposus in rabbit. The reason for the decreases in protein content remains unclear. METHODS: Anulus fibrosus and nucleus pulposus were cultured for 72 hours with or without chondroitinase ABC stimulated or not stimulated by interleukin-1 after preculture for 4 days. Subsequently, the MMPs (gelatinases MMP-2, MMP-9, and collagenase) and inflammatory mediators (prostaglandin E2 and nitric oxide) produced in the culture media were analyzed. RESULTS: In the anulus fibrosus chondroitinase ABC and interleukin-1 synergistically increased the collagenase activity, which was at a significantly higher level than the increment solely due to interleukin-1. In contrast, chondroitinase ABC counteracted the increase in nitric oxide production by interleukin-1. In the nucleus pulposus the collagenase and nitric oxide productions were not particularly affected by chondroitinase ABC and/or interleukin-1. In zymographic analysis MMP-2 was detected, but MMP-9 was only slightly detected in both tissues. There were no significant differences in both tissues for MMP-2 and prostaglandin E2 following incubation with or without chondroitinase ABC, whether stimulated by interleukin-1 or not. CONCLUSIONS: The collagenase activity in the anulus fibrosus was increased by chondroitinase ABC with interleukin-1. This finding may support the hypothesis that some proteolytic activities are involved in the chemonucleolytic process by chondroitinase ABC treatment. PMID- 11884905 TI - Dexamethasone decreases blood flow in normal nerves and dorsal root ganglia. AB - STUDY DESIGN: An experimental physiologic and histologic study of dexamethasone effects on peripheral nerves. OBJECTIVE: To characterize the effect of topically applied 0.4% dexamethasone on acute changes in nerve blood flow and subsequent histologic changes in rat sciatic nerve fibers. SUMMARY OF BACKGROUND DATA: Dexamethasone is an anti-inflammatory glucocorticoid used clinically to reduce the neural consequences of inflammation. Several reports of accidental injury to nerves after steroid injections have raised questions about the mechanisms involved in dexamethasone-induced neurotoxic injury. METHODS: Nerve blood flow studies using a laser Doppler flowmeter were conducted in animals with stable temperature and arterial pressure. Dexamethasone 0.4%, 0.1 mL was applied topically to rat sciatic nerve in the following protocol groups: 1) nerve blood flow recording every 5 minutes for 30 minutes, and 2) initial nerve blood flow recording and repeat recording at 4 hours. Three additional animals had 30-minute nerve blood flow recordings in which normal saline was substituted for dexamethasone; these animals were used for control and to assure that the experimental preparation was viable throughout the observational period. Additional groups of two animals each received dexamethasone but were used only for neuropathologic observation at 2, 4, and 6 days after treatment. Neuropathologic studies were conducted on glutaraldehyde-fixed, plastic-embedded tissue. RESULTS: Application of saline to the exposed sciatic nerves did not significantly change nerve blood flow from baseline values. Nerve blood flow values remained constant throughout the observational period. Dexamethasone, however, significantly reduced nerve blood flow in both the 30-minute and 4-hour groups. Some animals showed an initial transient increase in blood flow before nerve blood flow began to steadily decline to the final values reported. Neuropathologic changes were minimal and consisted only of edema and occasional subperineurial activation of Schwann cells. No demyelination or degeneration was seen. CONCLUSION: Dexamethasone causes statistically significant reductions in normal nerve blood flow at 30 minutes and 4 hours after topical application; however, the reduction is on average below the threshold for causing ischemic changes in the structure of peripheral nerve fibers. PMID- 11884906 TI - Neural space integrity of the lower cervical spine: effect of normal range of motion. AB - STUDY DESIGN: An experimental investigation of intervertebral foramen and spinal canal neural space integrity was performed throughout physiologic range of motion of the lower cervical spine in intact human cadaver specimens. OBJECTIVE: To investigate cervical positions that might place the neural tissues of the spine in heightened risk of injury. To meet this objective the following hypotheses were tested: 1) spinal canal integrity varies with specific normal range of motion positions of the lower cervical spine, and 2) intervertebral foramen integrity is dependent on and unique for different physiologic positions of the lower cervical spine. SUMMARY OF BACKGROUND DATA: Cervical spine injuries are frequently associated with compressive damage to neurologic tissues and consequently poor clinical outcomes. Neurologic injury typically occurs from disc, ligamentous, or bony occlusion of the spinal canal and intervertebral foraminal spaces dynamically during an injury event or with abnormal alignment and position after the injury event. Prior studies have shown pressure and geometric changes in cervical spine neural spaces in certain cervical spine positions. However, to the authors' knowledge, this is the first research effort aimed at elucidating the integrity of the cervical spine neural spaces throughout the normal physiologic range of motion. METHODS: The authors instrumented 17 fresh-frozen unembalmed cadaveric human cervical spines (C3-C7) with specially designed intervertebral foramen occlusion transducers and a spinal canal occlusion transducer. The specimens were loaded with pure bending moments to produce simulated physiologic motions of the lower cervical spine. The resulting occlusion profiles for the intervertebral foramen and spinal canal were recorded along with the 6-degree of freedom position of the cervical spine. Because these occlusion measurements describe the ability of the spine to preserve the space for the neural structures, the authors define this neuroprotective role of the vertebral column as neural space integrity. RESULTS: The range of motion developed experimentally in this study compared well with published reports of normal cervical motion. Thus, subsequent changes in neural space integrity may be regarded as resulting from normal human cervical spine motion. No significant change in the spinal canal space was detected for any physiologic motion; however, intervertebral foramen integrity was significantly altered in extension, ipsilateral bending, combined ipsilateral bending and extension, and combined contralateral bending with extension when compared with intact upright neutral position. CONCLUSIONS: This study defines the range of neural space integrity associated with simulated physiologic motion of the lower cervical spine in an experimental setting. This information may be useful in comparing neural space changes in pathologic conditions and may enhance refinement of neurologic injury prevention strategies. PMID- 11884907 TI - Efficacy of the transcutaneous electrical nerve stimulation for the treatment of chronic low back pain: a meta-analysis. AB - BACKGROUND: Low back pain affects a large proportion of the population. Transcutaneous electrical nerve stimulation (TENS) was introduced more than 30 years ago as an alternative therapy to pharmacologic treatments for chronic pain. However, despite its widespread use, the efficacy of TENS is still controversial. PURPOSE: The aim of this meta-analysis was to determine the efficacy of TENS in the treatment of chronic low back pain. METHODS: The authors searched MEDLINE, EMBASE, PEDro, and the Cochrane Controlled Trials Register up to June 1, 2000. Only randomized controlled clinical trials of TENS for the treatment of patients with a clinical diagnosis of chronic low back pain were included. Abstracts were excluded unless further data could be obtained from the authors. Two reviewers independently selected trials and extracted data using predetermined forms. DATA ANALYSIS: Heterogeneity was tested with Cochrane's Q test. A fixed effects model was used throughout for continuous variables, except where heterogeneity existed, in which case, a random effects model was used. Results are presented as weighted mean differences with 95% confidence intervals, where the difference between the treated and control groups was weighted by the inverse of the variance. Standardized mean differences were calculated by dividing the difference between the treated and control by the baseline variance. Standardized mean differences were used when different scales were integrated to measure the same concept. Dichotomous outcomes were analyzed with odds ratios. MAIN RESULTS: Five trials were included, with 170 subjects randomized to the placebo group receiving sham TENS and 251 subjects receiving active TENS (153 for conventional mode, 98 for acupuncture-like TENS). The schedule of treatments varied greatly between studies ranging from one treatment/day for 2 consecutive days, to three treatments/day for 4 weeks. There were no statistically significant differences between the active TENS group compared with the placebo TENS group for any outcome measures. Subgroup analysis performed on TENS application and methodologic quality did not demonstrate a significant statistical difference (P > 0.05). Remaining preplanned subgroup analysis was not conducted because of the small number of included trials and the variety of outcome measures reported. CONCLUSION: The results of the meta-analysis present no evidence to support the use or nonuse of TENS alone in the treatment of chronic low back pain. Considering the small number of studies responding to the criteria to be included in this meta-analysis, it is clear that more appropriately designed studies are needed before a final conclusion. Clinicians and researchers should consistently report the characteristics of the TENS device and the application techniques used. New trials on TENS should make use of standardized outcome measures. This meta analysis lacked data on how TENS efficacy is affected by four important factors: type of applications, site of application, treatment duration of TENS, and optimal frequencies and intensities. PMID- 11884909 TI - Clinical outcome results of pedicle subtraction osteotomy in ankylosing spondylitis with kyphotic deformity. AB - STUDY DESIGN: A prospective study was performed in 45 patients with ankylosing spondylitis. OBJECTIVES: To assess the outcomes of decancellation pedicle subtraction extension osteotomy in ankylosing spondylitis patients with severe fixed kyphotic deformity. SUMMARY OF BACKGROUND DATA: There have been several studies regarding correction of kyphotic deformity in ankylosing spondylitis. However, most of them concern surgical technique. There have been no reports concerning clinical results of decancellation pedicle subtraction osteotomy in ankylosing spondylitis. METHODS: The kyphotic deformity was corrected by a one stage pedicle subtraction extension osteotomy. Radiographic assessment for sagittal balance was performed by measuring thoracic kyphosis, lumbar lordosis, distance between the vertical line on anterosuperior point of T1 and that of S1, and sacral inclination. Chin brow-vertical angle was measured on the preoperative and postoperative clinical photograph of patients. Clinical outcomes were assessed by questionnaire measuring changes in physical function, indoor activity, outdoor activity, psychosocial activity, pain, and patient satisfaction with surgery. RESULTS: Final follow-up radiograph showed an increase in lumbar lordosis from 10 degrees to 44 degrees (an increase of 34 degrees), whereas thoracic kyphosis remained stable from 50 degrees to 54 degrees. Sagittal imbalance significantly improved from 94 to 8 mm, whereas sacral inclination increased from 8 degrees to 24 degrees. The chin brow-vertical angle was 32.0 degrees before surgery and 0.9 degrees after surgery. Satisfactory clinical outcome was achieved; however, clinical improvements did not correlate with changes in radiologic measurements. CONCLUSIONS: Most of the patients maintained good correction and had good clinical results. Based on the results of this study, pedicle subtraction extension osteotomy is effective for correction of kyphotic deformity in ankylosing spondylitis. PMID- 11884908 TI - Curve prevalence of a new classification of operative adolescent idiopathic scoliosis: does classification correlate with treatment? AB - STUDY DESIGN: A retrospective multicenter consecutive case review of operative adolescent idiopathic scoliosis. OBJECTIVES: To define the curve prevalence of a large consecutive series of cases with operative adolescent idiopathic scoliosis as classified by a new system and to test the ability of this new classification system to correlate with regions of the scoliotic spine to be instrumented/fused. SUMMARY OF BACKGROUND DATA: A new comprehensive, two-dimensional classification system, intended to be treatment based, has been developed. However, it has not been tested whether all presenting operative cases of adolescent idiopathic scoliosis are classifiable in a large consecutive series, nor has the prevalence of specific curve types been determined. In addition, it is unknown whether this classification is truly treatment based, as to whether it can correlate with regions of the spine to be instrumented/fused. METHODS: A multicenter retrospective review of 606 consecutive operative cases of adolescent idiopathic scoliosis was performed. All cases were classified by a new triad classification system, which included the following: a curve type (1-6), a lumbar spine modifier (A, B, C), and a sagittal thoracic modifier (-, N, +). Prevalence of the individual three components of the system and the classification grouping of all three components together were performed. In addition, the authors assessed whether this system could correlate with regions of the spine that should be included in the instrumentation and fusion, based on exactly which regions were fused during the operative procedure. RESULTS: All 606 cases were classifiable by this system. Prevalence of the six curve types noted was as follows: Type 1, main thoracic (n = 305, 51%); Type 2, double thoracic (n = 118, 20%); Type 3, double major (n = 69, 11%); Type 4, triple major (n = 19, 3%); Type 5, thoracolumbar/lumbar (n = 74, 12%); and Type 6, thoracolumbar/lumbar-main thoracic (n = 17, 3%). The five most common curve classifications noted were as follows: 1AN, 1BN, 2AN, 5CN, and 1CN, which accounted for 58% of all curve classifications noted. An average of 90% of the operative cases had surgically structural regions of the spine included in the instrumentation and fusion as predicted by the curve type. CONCLUSIONS: A new comprehensive classification system for operative adolescent idiopathic scoliosis found all 606 consecutive cases of adolescent idiopathic scoliosis classifiable, with the Type 1, main thoracic curve pattern, the most common curve type found (51%). This new classification system appears to correlate with treatment of surgically structural regions of the spine fused in 90% of cases by the objective radiographic criteria used. PMID- 11884910 TI - Long-term results from in situ fusion for congenital vertebral deformity. AB - STUDY DESIGN: Retrospective review of long-term outcome of fusion in situ for congenital vertebral anomaly with particular emphasis on cosmesis and the incidence of reoperation. OBJECTIVE: Examination of the success rate of this procedure and of risk factors for failure. BACKGROUND: Fusion in situ is the accepted prophylactic treatment to prevent deformity in congenital vertebral anomalies that have a high risk of progression or have been shown to be deteriorating. METHODS: Records of patients who were at least 15 years of age at last examination were reviewed retrospectively. Consideration was given to cosmetic outcome and to the incidence of reoperation. RESULTS: There were 43 patients in this category, 19 boys and 24 girls, who were at least 15 years of age when last seen. Reoperation had been performed in 11 cases (25.6%). The main finding was that, although the Cobb angle of the fused segment of spine remained constant after fusion, a curve sometimes developed in the whole spine, sometimes (but by no means always) centered on that fused segment. Cosmetic deformity continued to progress in a number of cases. CONCLUSIONS: Localized fusion, whether posterior alone or anterior and posterior combined, was effective in preventing progression of the Cobb angle of the congenitally malformed area but did not control the overall deformity that developed or progressed with growth. Current concepts of the pathomechanism of deformity do not adequately explain the observations, and a more biologic approach is suggested. PMID- 11884911 TI - Some complications of common treatment schemes of thoracolumbar spine fractures can be predicted with magnetic resonance imaging: prospective study of 53 patients with 71 fractures. AB - STUDY DESIGN: Prospective cohort study. OBJECTIVES: To study the predictive value of magnetic resonance imaging (MRI) findings of thoracolumbar spine fractures concerning the radiologic and clinical outcome. SUMMARY OF BACKGROUND DATA: Disagreement about the proper treatment of thoracolumbar spine fractures is caused by insufficiency of conventional imaging techniques. Previous studies have shown that MRI is capable of distinguishing injury to all structures of the fractured spine and thus may help develop schemes with higher predictive power. METHODS: A total of 53 patients with 71 fractures were studied with MRI in a prospective fashion. A total of 24 patients with 39 fractures were treated conservatively and 29 patients with 32 fractures were treated operatively after a protocol concerning the treatment options. MRI scans were obtained within 1 week of injury and at the 2-year follow-up. Pain scores were obtained at the 2-year follow-up. Previously described MRI schemes concerning the trauma and post-trauma conditions were used. RESULTS AND CONCLUSIONS: An unfavorable outcome in the conservative group was related to the progression of kyphosis, which in most cases was predictable with the use of trauma MRI findings concerning the endplate comminution and vertebral body involvement. In the operatively treated group, recurrence of the kyphotic deformity was predictable by the lesion of the posterior longitudinal ligamentary complex together with endplate comminution and vertebral body involvement as seen on trauma MRI. The authors recommend the use of MRI to develop reliable prognostic criteria for these injuries. PMID- 11884912 TI - Fatigue-related changes in torque output and electromyographic parameters of trunk muscles during isometric axial rotation exertion: an investigation in patients with back pain and in healthy subjects. AB - STUDY DESIGN: A cross-sectional case-control study. OBJECTIVES: To examine the effect of fatigue on torque output as well as electromyographic frequency and amplitude values of trunk muscles during isometric axial rotation exertion in back pain patients and to compare the results with a matched control group. SUMMARY OF BACKGROUND DATA: Back pain patients exhibited different activation strategies in trunk muscles during the axial rotation exertions. Fatigue changes of abdominal and back muscles during axial rotation exertion have not been examined in patients with back pain. METHODS: Twelve back pain patients and 12 matched controls performed isometric fatiguing axial rotation to both sides at 80% maximum voluntary contraction in a standing position. During the fatiguing exertion, electromyographic changes of rectus abdominis, external oblique, internal oblique, latissimus dorsi, iliocostalis lumborum, and multifidus were recorded bilaterally. The primary torque in the transverse plane and the coupling torques in sagittal and coronal planes were also measured. RESULTS: No difference in the endurance capacity was found between back pain and control groups. At the initial period of the exertion, back pain patients demonstrated a statistical trend (P = 0.058) of greater sagittal coupling torque as well as lower activity of rectus abdominis and multifidus and higher activity in external oblique. During the fatigue process similar changes of coupling torque were demonstrated in both sagittal and coronal planes, but a smaller fatigue rate for right external oblique, increase in median frequency for latissimus dorsi, and lesser increase in activity for back muscles were found in the back pain group compared with the control group. CONCLUSIONS: Alterations in electromyographic activation and fatigue rates of abdominal and back muscles demonstrated during the fatigue process provide insights into the muscle dysfunctions in back pain and may help clinicians to devise more rational treatment strategies. PMID- 11884913 TI - Quality of life and cost of care of back pain patients in Finnish general practice. AB - STUDY DESIGN: A cohort of 114 primary care patients were studied for 1 year before and 1 year after a randomized clinical trial. OBJECTIVES: To explore the therapy use, societal costs, and quality of life of patients with prolonged back pain. To compare the effects of physiotherapy, bone setting, and light exercise therapy on these measures. SUMMARY OF BACKGROUND DATA: Analyses of back pain have mostly focused on the minority of patients who cause high costs and a heavy burden on national economies. The majority with low costs have aroused less interest. The patient's choice of therapy, especially alternative medicine, has seldom been evaluated despite the increasing popularity of alternative therapies. METHODS: Data were collected from the Social Insurance Institution files, patient records, and questionnaires: the Nottingham Health Profile (NHP). RESULTS: One year before enrollment a third of the patients had consulted primary care. Half of them had had some therapy: mainly massage, physiotherapy, naprapathy, or bone setting. One third of the direct costs were spent on complementary therapies and another third on rehabilitation. Sick leaves accounted for 55% of the total costs (US$ 1029). The mean total costs slightly increased after the randomized therapies (US$ 1306). The costs of ambulatory care, with the study therapies included, were similar, whereas physiotherapy seemed the cheapest (US$ 621) and bone setting the most expensive (US$ 2072) alternative in view of the total costs. More NHP subscales were improved by physiotherapy and bone setting than by exercise. CONCLUSIONS: A third of the direct back pain costs were spent on complementary therapies. The use of health care services and absenteeism tended to decrease after a course of physiotherapy. Physiotherapy and bone setting seemed able to improve the quality of life of patients with prolonged back pain. PMID- 11884914 TI - Active sick leave for patients with back pain: all the players onside, but still no action. AB - STUDY DESIGN: Semistructured interviews, group discussions, and a mailed survey. OBJECTIVE: To identify barriers to the use of active sick leave (ASL) and to design an intervention to improve the use of ASL by patients with low back pain. SUMMARY OF BACKGROUND DATA: ASL was introduced in Norway in 1993 to encourage people on sick leave to return to modified work. With ASL the National Insurance Administration (NIA) pays 100% of wages, thereby allowing the employer to engage a substitute worker at no extra cost, in addition to the worker on ASL. Arranging ASL requires cooperation between the general practitioner (GP), employer, local NIA staff, and the patient, which may explain why ASL was used in less than 1% of the eligible sick leave cases in 1995, despite strong support from all players. METHODS: The authors conducted five in-depth interviews at a workplace where ASL was successfully implemented. Questionnaires were sent to 89 GPs, 102 workplace representatives, and 22 local NIA officers in three counties. Five patients with back pain who had used ASL were interviewed in a focus group, and 10 patients with back pain who had not used ASL were interviewed using a structured guide. Five workplaces participated in a dialogue conference. Data collection and analysis were iterative, and new data were constantly compared with the previously analyzed materials. RESULTS: About 80% of the GPs, employers, and NIA officers believed ASL is effective in reducing long-term sick leave. Among the barriers identified were lack of information, lack of time, and work flow barriers such as poor communication and coordination of activities between the players required to carry out ASL. Two strategies were designed to improve the workflow between them. A passive implementation strategy was designed to require a minimum amount of economic and administrative support. It included targeted information, clinical guidelines for low back pain, a reminder to GPs in the sick leave form, and a standardized agreement. A proactive strategy included the same four elements plus a kick-off continuing education seminar for GPs and a trained resource person to facilitate the use of ASL. CONCLUSIONS: Having all the players onside may be essential, but it is not sufficient to bring about action in workplace strategies for patients with low back pain. If early return to modified work is effective, implementing it may require interventions targeted at identified barriers. PMID- 11884915 TI - Physical, psychosocial, and individual risk factors for neck/shoulder pain with pressure tenderness in the muscles among workers performing monotonous, repetitive work. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVES: To evaluate the effect of individual characteristics and physical and psychosocial workplace factors on neck/shoulder pain with pressure tenderness in the muscles. SUMMARY OF BACKGROUND DATA: Controversy prevails about the importance of workplace factors versus individual factors in the etiology of pain in the neck and/or shoulders. METHODS: Study participants were 3123 workers from 19 plants. Physical risk factors were evaluated via video observations, and psychosocial risk factors were assessed with the job content questionnaire. Other procedures included symptom survey, clinical examination, and assessment of health-related quality of life (SF-36). The main outcome variable, neck/shoulder pain with pressure tenderness, was defined on the basis of subjective pain score and pressure tenderness in muscles of the neck/shoulder region. RESULTS: The prevalence of neck/shoulder pain with pressure tenderness was 7.0% among participants performing repetitive work and 3.8% among the referents. We found an association with high repetitiveness (prevalence ratio 1.8, 95% confidence interval 1.1-2.9), high force (2.0, 1.2 3.3), and high repetitiveness and high force (2.3, 1.4-4.0). The strongest work related psychosocial risk was high job demands (1.8, 1.2-2.7). Increased risk was also associated with neck/shoulder injury (2.6, 1.6-4.1), female gender (1.8, 1.2 2.8), and low pressure pain threshold (1.6, 1.1-2.3). Neck/shoulder pain was strongly associated with reduced health-related quality of life. CONCLUSIONS: Work-related physical and psychosocial factors, as well as several individual risk factors, are important in the understanding of neck/shoulder pain. The findings suggest that neck/shoulder pain has a multifactorial nature. Reduced health-related quality of life is associated with subjective pain and clinical signs from the neck and shoulders. The physical workplace factors were highly intercorrelated, and so the effect of individual physical exposures could only be disentangled to a minor degree. PMID- 11884917 TI - Flail chest secondary to excessive rib resection in idiopathic scoliosis: case report. AB - STUDY DESIGN: Case report. OBJECTIVES: To report a previously undescribed complication of scoliosis surgical treatment. SUMMARY OF BACKGROUND DATA: None available. METHODS: Clinical case analysis. RESULTS: Excessive rib resection resulted in a permanent "flail" chest. CONCLUSION: Rib resection ("costoplasty") is a valuable procedure for obtaining bone graft and for esthetic reduction of rib prominence, but excessive removal and especially done twice can produce major disability. PMID- 11884919 TI - Is Cobb angle progression a good indicator in adolescent idiopathic scoliosis? AB - STUDY DESIGN: A retrospective follow-up study of spine geometry after posterior instrumentation and fusion for adolescent idiopathic scoliosis (AIS). OBJECTIVES: To evaluate 1) if Cobb angle progression is a reliable indicator of the crankshaft phenomenon; 2) if significant growth of the spine can occur after surgery without the development of a crankshaft phenomenon? SUMMARY OF BACKGROUND DATA: Anterior fusion of the spine is often recommended for skeletally immature scoliotic patients to avoid the risk of a crankshaft phenomenon, a long-term loss of curve correction caused by residual growth of the spine combined with the constraints of a posterior fusion. The crankshaft phenomenon is usually assessed indirectly by documenting progression of the Cobb angle on frontal radiographs. Thus far, no study has directly measured the three-dimensional growth of the spine after surgery in AIS. METHODS: Cobb angle, spine length and spine height were obtained from three-dimensional radiographic reconstructions of the spine in 48 adolescent scoliotic patients undergoing posterior instrumentation and fusion. Measurements were done before surgery, after surgery and at skeletal maturity. A significant growth of the spine was defined as a > or = 10 mm increase in spine length, while a significant curve progression was defined as a > or = 10 degrees increase in Cobb angle at skeletal maturity. RESULTS: In the majority of patients (56%), there was no significant change in spinal length or in Cobb angle measurements at an average 2.4 years post surgery. A crankshaft phenomenon was detected in 6 patients (12%) for which significant increases both in spinal length and Cobb angle measurement were found. Significant curve progression without any change in spine length was noted in 9 patients (19%) while an increase in spine length with no evidence of curve progression was present in 6 patients at last follow-up. CONCLUSION: Spinal growth as indicated by an increase in spinal length can be measured in a significant proportion of adolescents with idiopathic scoliosis after posterior instrumentation and fusion. Some of these study participants will develop a crankshaft phenomenon but Cobb angle progression is not a reliable indicator of this complication, since it may occur without any detectable growth of the spine. PMID- 11884920 TI - Diverging intramuscular activity patterns in back and abdominal muscles during trunk rotation. AB - STUDY DESIGN: An intramuscular electromyographic study was performed on trunk rotations during sitting and standing. OBJECTIVE: The aim was to provide new information on activation levels for deep trunk muscles in various unresisted and resisted trunk rotations. SUMMARY AND BACKGROUND DATA: Frequent daily trunk twisting and decreased maximal strength during trunk rotation have been associated with low back pain or sciatic pain. However, the involvement of deep trunk muscles during different trunk rotations is relatively unknown. METHODS: Ten healthy subjects participated. Fine-wire electrodes were inserted, under ultrasound guidance, into psoas, quadratus lumborum, the superficial medial lumbar erector spinae (ES-s, multifidus) and its deep lateral portion (ES-d, iliocostalis), iliacus, rectus abdominis, obliquus externus, and obliquus internus. RESULTS: The highest involvement for all muscles was observed on the ipsilateral side, in maximal trunk twists with shoulder resistance, except obliquus externus, which showed a dominant contralateral side, and rectus abdominis, which was little activated in all rotations. In contrast, maximal trunk twist without shoulder resistance, i.e., freely performed, resulted generally in lower levels for all muscles involved and in a shift of side dominance for the lumbar muscles quadratus lumborum, psoas, and ES-s. CONCLUSIONS: During trunk rotations the activity patterns for various trunk muscles could drastically change, and even be the opposite, between the two body sides, within the same type of task, depending on several factors such as initial position, effort level, sitting or standing, and external shoulder resistance. PMID- 11884921 TI - Partial lumbosacral kyphosis reduction, decompression, and posterior lumbosacral transfixation in high-grade isthmic spondylolisthesis: clinical and radiographic results in six patients. AB - BACKGROUND: In an attempt to increase fusion while decreasing the neurologic risk associated with complete reduction of high-grade spondylolisthesis, the authors have used a technique of partial lumbosacral kyphosis reduction, posterior decompression, and pedicle screw transfixation of the lumbosacral junction. OBJECTIVE: To determine if this technique is effective in treatment of high-grade spondylolisthesis. STUDY DESIGN: A retrospective review of six patients with high grade spondylolisthesis treated by this technique was performed. There were four female patients (ages 16 years [n=2], 23 years [n=1], and 29 years [n=1]) and two male patients (both 13 years of age) with spondylolisthesis ranging from Grade IV to Grade V. All patients presented with pain and radiculopathy. After surgery the patients were evaluated for resolution of symptoms, sagittal alignment, fusion, and satisfaction. The radiographic measurements included the slip angle, the percentage slip, and the sacral inclination. An SRS outcome score was also obtained on all six patients to evaluate postoperative outcome, in terms of pain control, self-image perception, and return to function. RESULTS: The average length of follow-up was 42.6 months (range 24-60 months). All patients evidenced solid fusion by the 6-month follow-up (based on oblique radiographs showing lateral bridging bone masses). The slip angle was improved from 62 degrees to 28 degrees (P < 0.5), whereas there was no significant improvement in the percentage slip or the sacral inclination (89-80% and 28-37 degrees, respectively). No progression of the slip angle or percentage slip was noted on the follow-up radiographs. Complications included two intraoperative dural tears that were identified and repaired. There were no neurologic complications. The SRS outcome instrument demonstrated good postoperative pain control, function, self-image, and satisfaction in all patients. CONCLUSION: In high-grade spondylolisthesis, this posterior approach is safe and effective in obtaining a solid arthrodesis, restoring sagittal balance, and improving function. These results reinforce the impression that it is the partial reduction of the slip angle, not the percentage slip, in high-grade spondylolisthesis that is important in obtaining optimal results. PMID- 11884922 TI - Extraforaminal entrapment of the fifth lumbar spinal nerve by osteophytes of the lumbosacral spine: anatomic study and a report of four cases. AB - STUDY DESIGN: An anatomic study of the associations between the fifth lumbar spinal nerve (L5 spinal nerve) and a lumbosacral tunnel, consisting of the fifth lumbar vertebral body (L5 vertebral body), the lumbosacral ligament, and sacral ala, and clinical case reports of four patients with lumbar radiculopathy secondary to entrapment of the L5 spinal nerve in the lumbosacral tunnel. OBJECTIVES: To delineate the anatomic, clinical, and radiologic features and surgical outcome of patients with entrapment of the L5 spinal nerve in the lumbosacral tunnel. SUMMARY OF BACKGROUND DATA: Although several cadaveric studies on a lumbosacral tunnel as a possible cause of L5 radiculopathy have been reported, few studies had focused on osteophytes of the L5-S1 vertebral bodies as the major component of this compressive lesion, and clinical reports on patients with this disease have been rare. METHODS: Lumbosacral spines from 29 geriatric cadavers were examined with special attention to the associations between osteophytes of the L5-S1 vertebral bodies and the L5 spinal nerve. Four patients with a diagnosis of the entrapment of the L5 spinal nerve by osteophytes at the lumbosacral tunnel were treated surgically, and their clinical manifestations and surgical results were reviewed retrospectively. RESULTS: The anatomic study demonstrated osteophytes of the L5-S1 vertebral bodies in seven of the 29 cadavers. Entrapment of the L5 spinal nerve in the lumbosacral tunnel was observed in six of the seven cadavers with L5-S1 osteophytes but in only one of the 22 cadavers without such osteophytes (P < 0.05, chi2 test). All four patients had neurologic deficits in the L5 nerve root distribution. MRI and myelography showed no abnormal findings in the spinal canal, but CAT scans demonstrated prominent osteophytes on the lateral margins of L5-S1 vertebral bodies in all four. Selective L5 nerve block completely relieved all patients of pain but only temporarily. Three patients were treated via a posterior approach by resecting the sacral ala along the L5 spinal nerve, and the other patient was treated by laparoscopic anterior resection of the osteophytes. Pain relief was obtained in the four patients immediately after surgery, but one patient experienced recurrence of pain 1 year after the first surgery and was successfully treated by additional posterior decompression and fusion. CONCLUSIONS: Extraforaminal entrapment of L5 spinal nerve in the lumbosacral tunnel can cause L5 radiculopathy, and osteophytes of L5-S1 vertebral bodies are a major cause of the entrapment. PMID- 11884923 TI - Subdural hematoma after cervical epidural steroid injection. AB - STUDY DESIGN: A case report is presented involving a subdural hematoma after cervical epidural steroid injection. OBJECTIVE: To demonstrate a previously unreported complication of cervical epidural steroid injection. SUMMARY OF BACKGROUND DATA: Cervical epidural steroid injection is a common procedure performed in the care of patients with spine-related complaints. Reports of complications are rare, and most of these are fairly benign. To the authors' knowledge, subdural hematoma has never been described as a complication of a cervical epidural steroid injection. METHODS: A patient underwent an uncomplicated cervical epidural steroid injection by an experienced anesthesiologist. She developed acute onset of axial pain followed by progressive quadriparesis within a matter of 8 hours. She was transferred from a local emergency room after a CT scan suggested posterior cord displacement consistent with an anterior spinal hematoma from C3 to C5. She was taken to the operating room for urgent decompression. Exploration revealed an anterior subdural hematoma that was evacuated followed by dural closure with a patch. RESULTS: After surgery the patient was initially quadriplegic but rapidly gained full function in the left upper and lower extremities. She was making steady progress with motor recovery on the right side when she developed acute meningitis about 8 days after surgery, and then she subsequently went into cardiopulmonary arrest. She was successfully resuscitated but remained critically ill with no evidence of encouraging neurologic function. Six days later she had a second cardiac arrest and could not be resuscitated. CONCLUSIONS: It is important to acknowledge that spinal hematomas can occur after cervical epidural steroid injection, as prompt recognition and treatment could improve the prognosis for recovery. The sequelae of a cervical subdural hematoma after epidural steroid injection remain potentially devastating. PMID- 11884924 TI - Hepatic steatosis and liver transplantation current clinical and experimental perspectives. PMID- 11884925 TI - Hepatitis B core antibody-positive liver recipients and hepatitis B reaction after liver transplantation. PMID- 11884926 TI - Further lessons from knockout man. PMID- 11884927 TI - Feasibility of immunosuppression in composite tissue allografts by systemic administration of CTLA4Ig. AB - BACKGROUND: Although recent experimental studies have demonstrated CTLA4Ig to be a potent immunosuppressant in vascularized solid organ allografts, little attention has been given to the effect of this soluble recombinant fusion protein on immunosuppression in composite tissue allografts (CTAs). Using a rat hind limb allograft model, we examined the efficacy of CTLA4Ig against the allograft rejection of composite tissue. METHODS: The hind limbs of ACI rats (RT1a) were heterotopically transplanted to Lewis rats (RT11). Controls received no immunotherapy. Experimental recipients were treated with a single i.p. injection of either human immunoglobulin (Ig)G (0.5 mg/body) or CTLA4Ig (0.5 mg/body) according to different time schedules. Graft survival time and histopathological changes for each experimental group were evaluated and statistically compared. RESULTS: Graft survival times were prolonged significantly in rats treated with CTLA4Ig on day 1 and day 2 after transplantation, compared with survival times of controls. In particular, the most significant prolongation was found in rats treated on day 2. At 7 days after transplantation, moderate-to-severe histological rejection occurred in all tissues in control rats. On the other hand, in rats treated with CTLA4Ig, all tissues showed significantly better preservation. Among these treated rats, the rats treated on day 2 showed excellent histopathological conditions in each tissue. CONCLUSIONS: This study supports the feasibility of using CTLA4Ig for preventing acute rejection in CTA. On the basis of the current results, the administration of CTLA4Ig for CTA is more effective at 24-48 hr after transplantation, after the initial immune response has been allowed to begin. PMID- 11884928 TI - Role of hypomagnesemia in chronic cyclosporine nephropathy. AB - BACKGROUND: Hypomagnesemia is a common finding of cyclosporine (CsA)-treated patients and has been proposed as both a cause and a consequence of CsA-induced nephrotoxicity. This experiment was conducted to elucidate the role of hypomagnesemia in the pathogenesis of chronic CsA nephropathy. METHODS: CsA (15 mg/kg/day subcutaneously) was administered to rats maintained on a low-sodium diet for 1, 2, and 4 weeks, and the effects of magnesium (Mg) supplementation on renal function, renal histology, and renal gene expression profile of fibrogenic molecules and vasoconstrictors was examined. RESULTS: CsA elicited hypomagnesemia and induced a progressive decline in glomerular filtration. At 28 day, renal tubular atrophy and cortical striped interstitial fibrosis were evident with CsA treatment. Dietary supplementation of Mg ameliorated CsA-induced hypomagnesemia and almost completely abolished CsA-induced chronic fibrotic lesions. Neither CsA nor Mg supplementation affected blood pressure. Renal cortical mRNA of transforming growth factor beta, plasminogen activator inhibitor (PAI)-1, and extracellular matrix started to increase at 14 days and elevated further at 28 days. In contrast, the increase in mRNA of tissue inhibitor of matrix metalloproteinase-1 and renin was evident early at 7 days and reached peak at 14 days. These mRNA increases, except that of renin, were almost abolished when hypomagnesemia was corrected. Magnesium supplementation also improved glomerular dysfunction, at least in part, through inhibition of up-regulated mRNA of endothelin-1. CONCLUSION: CsA-induced hypomagnesemia contributes to chronic renal fibrotic lesions seen during CsA treatment through up-regulation of fibrogenic molecules, most notably early activation of tissue inhibitor of matrix metalloproteinase-1 expression. PMID- 11884929 TI - SP-A-enriched surfactant for treatment of rat lung transplants with SP-A deficiency after storage and reperfusion. AB - BACKGROUND: The function of pulmonary surfactant is affected by lung transplantation, contributing to impaired lung transplant function. A decreased amount of surfactant protein-A (SP-A) after reperfusion is believed to contribute to the impaired surfactant function. Surfactant treatment has been shown to improve lung transplant function, but the effect is variable. We investigated whether SP-A enrichment of surfactant improved the efficacy of surfactant treatment in lung transplantation. METHODS: Left and right lungs of Lewis rats, inflated with 50% O2, were stored for 20 hr at 8 degrees C. Surfactant in bronchoalveolar lavage fluid from right lungs was investigated after storage (n=6). Left lungs were transplanted into syngeneic recipients and treated with SP A-deficient surfactant (n=6) or SP-A-enriched surfactant (n=6) just before reperfusion. Air was instilled into untreated lung transplants (n=6). Sham operated (n=4) and normal (n=8) animals served as controls. Lung function was measured during 1 hr of reperfusion; surfactant components in bronchoalveolar lavage fluid were measured after reperfusion. RESULTS: After storage the amount of SP-A decreased by 27%, whereas surfactant phospholipids changed minimally. After reperfusion a further decrease of SP-A was paralleled by profound changes in surfactant phospholipids. Lung transplant function, however, remained relatively good. After instillation of SP-A-enriched surfactant, PO2 values were reached that approximated sham control PO2 values, whereas after SP-A-deficient surfactant treatment, the PO2 values did not improve. CONCLUSION: Enrichment of surfactant with SP-A for treatment of lung transplants improves the efficacy of surfactant treatment. PMID- 11884930 TI - Inhibitory effects of immunosuppressive drugs on insulin secretion from HIT-T15 cells and Wistar rat islets. AB - Until recently, islet allotransplantation for type 1 diabetic patients has been largely unsuccessful. Previous pharmacologic studies of single drugs have suggested that one factor contributing to this poor success is toxicity of immunosuppressive drugs on transplanted islets. However, no comprehensive study of agents currently used for islet transplantation has been previously reported. Consequently, we exposed HIT-T15 cells and Wistar rat islets to various concentrations of five immunosuppressive agents for 48 and 24 hr, respectively, and measured glucose-stimulated insulin secretion during subsequent static incubations. Results are expressed as percent reduction of insulin secretion at the lower and upper limits, respectively, of plasma drug concentrations used in clinical transplantation compared with control (no drug exposure). Insulin secretion from HIT-T15 cells was significantly inhibited by 74% and 90% after exposure to methylprednisolone (P<0.05), 11% and 24% after exposure to cyclosporine (P<0.01), 60% and 83% after exposure to mycophenolate (P<0.05), 56% and 63% after exposure to sirolimus (P<0.001), and 10% and 20% after exposure to tacrolimus (P<0.001). Insulin secretion from Wistar rat islets was reduced by 0% and 48% after exposure to mycophenolate (P<0.001) and 20% and 31% after exposure to tacrolimus (P<0.05). No reduction in insulin secretion was observed from either HIT-T15 cells or rat islets after exposure to daclizumab. The results support the hypothesis that toxicity of certain immunosuppressive drugs on beta cell function plays a role in the poor success of islet allotransplantation. This is especially true of intrahepatically transplanted islets, which are exposed to higher portal concentrations of immunosuppressive agents. These findings support the use of low-dose immunosuppressive drug protocols in clinical islet transplantation. PMID- 11884931 TI - Experiences with leflunomide in solid organ transplantation. AB - BACKGROUND: Leflunomide (Arava), a drug widely used for treatment of rheumatoid arthritis, has a very promising background in experimental transplantation. Its activity in experimental models of chronic rejection, its synergy with calcineurin phosphatase inhibitors, and its inhibitory effects on herpes virus replication are compelling reasons to pursue its clinical evaluation in transplantation. We report the use of this drug over the past 3 years in various clinical situations. METHODS: A retrospective review was performed in 53 liver and kidney transplant recipients receiving Arava. A single-dose pharmacokinetic (PK) study was first performed in stable, renal transplant recipients, and an initially targeted serum level of 100 microg/mL (300 microM) was calculated to require a loading dose of 1200-1400 mg over a 7-day period. We correlate the appearance of toxicity with serum levels of active drug and review the outcomes in patients whose clinical condition required dose reductions of conventional immune suppressive drugs. RESULTS: Fifty-three patients received leflunomide from 5 days to more than 430 days, and 37 patients received the drug for more than 60 days. The primary toxicity was anemia in the renal transplant patients and elevation of liver enzymes in the liver transplant patients. At comparable oral doses, serum levels were substantially lower and anemia more common in patients with serum creatinine >3 mg/dL. In liver and renal recipients with serum creatinine <3 mg/dL, the drug was well tolerated and dose-limiting side effects occurred in less than 15% when drug serum levels were less than 80 microg/ml. Patients with serum creatinine >3 mg/dL often required serum levels of active drug reduced to <60 microg/mL. In 12 of 18 renal patients treated for 200 days or more, the dose of cyclosporine or Prograf was reduced by a mean of 38.5% and stopped in one patient. The prednisone dose was reduced by a mean of 25% in these same 13 patients. Cyclosporine or FK506 was stopped completely in four liver recipients and reduced by 65% in another patient. No evidence of acute rejection developed in any of these liver or kidney transplant patients. CONCLUSION: Leflunomide seems to possess substantial immune suppressive potency in renal and liver transplant recipients and may be safely dosed for more than 300 days. The data suggest that calcineurin phosphatase inhibitors and prednisone can be safely reduced in patients with serum levels of active drug above 50 microg/mL. Because of a wide inter-patient range of active metabolite terminal half-life (>300%), monitoring of serum levels would seem to be an important part of its evaluation. PMID- 11884932 TI - Proteinuria after injection of human focal segmental glomerulosclerosis factor. AB - BACKGROUND: Recurrent focal segmental glomerulosclerosis (FSGS) is heralded by proteinuria that may remit after treatment with plasmapheresis or immunoadsorption. Study of recurrent FSGS has been hampered by lack of an animal model that exhibits a pattern of proteinuria that mimics human disease. We have obtained a component of FSGS patient plasma (FSGS factor) that increases glomerular albumin permeability (P(alb)) in vitro and causes transient proteinuria in vivo. METHODS: Plasma fractions containing FSGS factor and comparable plasma fractions from normal donors were injected into normal male Sprague-Dawley rats. Urinary protein, albumin, and creatinine were measured at various time points. Additionally, plasma samples from test animals were collected after injection and tested for FS activity defined by increased P(alb). Finally, glomeruli were isolated from animals after injection and P(alb) of these glomeruli tested. RESULTS: Proteinuria and albuminuria were increased by 24 hr after injection with FSGS factor, and returned to baseline by 48 hr after injection. Injection with the same fraction of normal plasma had no effect on urinary protein. FSGS factor increased urinary protein in a dose-dependent manner. Serum collected from rats 15 or 60 min after injection with FSGS factor increased P(alb) of glomeruli in vitro, whereas serum collected 3 or more hours after injection had no effect. Glomeruli isolated from rats receiving injections with FSGS factor had increased in vitro P(alb) compared with glomeruli from rats injected with a fraction from normal plasma. CONCLUSIONS: We have demonstrated that a single injection of FSGS factor increases P(alb) and, causes transient albuminuria and proteinuria in rats. FS activity in the plasma of recipient rats is also transient. This is the first detailed description of the time course and dose-dependence of proteinuria caused by FSGS factor in an animal model. PMID- 11884933 TI - Immunological and virological effects of ribavirin in hepatitis C after liver transplantation. AB - BACKGROUND: Hepatitis C recurring after liver transplant may cause progressive liver dysfunction, and available treatment regimens are unsatisfactory. A better understanding of the mechanisms of action of drugs currently used to manage hepatitis C would be helpful. METHODS: In a pilot, uncontrolled clinical trial, we treated 12 patients with post-liver transplantation hepatitis C with 1000-1200 mg qd of ribavirin, given as a monotherapy. We measured the transaminases levels, the liver disease grading and staging scores, the intrahepatic interferon-gamma, tumor necrosis factor-alpha, interleukin (IL)-4 and IL-10 mRNA levels, the serum and liver hepatitis C virus (HCV) RNA titers, and the intrahepatic HCV envelope 2 protein staining score before and after 12 weeks of ribavirin monotherapy. RESULTS: Ribavirin induced a significant amelioration of the transaminases levels. This biochemical response was not associated with a distinct change in the intrahepatic T helper 1/T helper 2 cytokine mRNA profile. Furthermore, some histological parameters, such as the portal inflammation and the fibrosis scores, worsened significantly even in the short term. A slight, albeit not significant, decrease of serum HCV RNA level and intrahepatic HCV antigen staining score was observed. Intrahepatic genomic-strand (but not negative-strand) HCV RNA titer decreased significantly (P=0.024). CONCLUSIONS: Contrary to what is suggested by experimental data, administration of ribavirin alone to patients with recurrent hepatitis C after liver transplantation is not accompanied by a specific change of the intrahepatic interferon-gamma, tumor necrosis factor-alpha, IL-4, or IL-10 mRNA transcription profile. PMID- 11884934 TI - Posttransplant diabetes mellitus in kidney allograft recipients: incidence, risk factors, and management. AB - BACKGROUND: Posttransplant diabetes mellitus (PTDM), associated with the use of immunosuppressants, occurs at varying rates in kidney transplant recipients. METHODS: Five transplant centers conducted a retrospective review of 435 kidney recipients completing at least 6 months of follow-up to determine risk factors, incidence, and management strategies for posttransplant glucose intolerance. A distinction was made between hyperglycemia and diabetes. RESULTS: The incidence of PTDM was found to be 4.9%. Among tacrolimus-treated patients it was 5.7%, compared with 3.3% among cyclosporine-treated patients (P=0.453). Mean daily maintenance doses of prednisone and mycophenolate mofetil (MMF) were significantly lower in tacrolimus-treated patients. Significantly more tacrolimus treated patients were prednisone-free (9.0%/0%; P<0.001). Logistic regression analysis revealed that the absence of an antiproliferative agent correlated with the development of PTDM (odds ratio=3.56; P=0.01). CONCLUSIONS: Based on this study, we propose management guidelines specifically for glucose intolerance developing after renal transplantation. Maintenance of blood glucose levels within strict limits is recommended, and the contribution of immunosuppressive agents to the development of PTDM is accounted for. Gradual tapering of prednisone and tacrolimus is proposed for patients who develop PTDM but also bear minimal risk of rejection. Tapering and eventual withdrawal of insulin should be attempted once blood glucose levels normalize. Switching to the alternative calcineurin inhibitor should only be considered as a late intervention. Tacrolimus therapy should be considered even in patients at high risk for diabetes, because the benefit of reduced acute rejection incidence and severity, as demonstrated in other studies, outweighs the risk of PTDM. PMID- 11884935 TI - Ki67, E-cadherin, and p53 as prognostic indicators of long-term outcome after liver transplantation for metastatic neuroendocrine tumors. AB - BACKGROUND: Patients suffering from hepatic metastases of neuroendocrine tumors (NET) are potential candidates for orthotopic liver transplantation. Because recurrence rates are high and outcome is variable, prognostic indicators are required. The aim of our study was to identify predictors of long-term survival with a focus on the impact of tumor biology. METHODS: We retrospectively analyzed 19 patients who received an orthotopic liver graft for metastatic NET at the Medizinische Hochschule Hannover. Expression of Ki67, E-cadherin, and p53 was studied immunohistochemically in metastases of neuroendocrine tumors of the explanted livers. RESULTS: Patients were followed up to 146 months after liver transplantation. Six patients died during follow-up. The resulting 1-, 5-, and 10 year survival rates are 89%, 80%, and 50%, respectively. All deaths during long term follow-up were tumor-associated. Recurrence was diagnosed in 12 patients between 2 weeks and 48 months after liver transplantation. Three patients are without tumor recurrence more than 8 years after liver transplantation. Survival in the 5 patients with low Ki67 and regular E-cadherin staining was significantly better than in the 12 patients with high Ki67 or aberrant E-cadherin expression (7-year survival 100% vs. 0%, respectively, log rank P=0.007). p53 expression did not significantly improve prognostic accuracy. CONCLUSIONS: We conclude that analysis of Ki67 and E-cadherin expression may improve the identification of patients with a favorable prognosis after liver transplantation for metastatic neuroendocrine tumors. PMID- 11884937 TI - Acute thrombosis of renal transplant artery: graft salvage by means of intra arterial fibrinolysis. AB - BACKGROUND: Arterial thrombosis in a transplanted kidney is a serious complication that usually leads to graft loss. The purpose of our study was to evaluate intra-arterial fibrinolysis as a treatment of acute renal transplant artery thrombosis and to determine the maximum period of occlusion allowing a reasonable chance of graft salvage. METHODS AND RESULTS: Four patients underwent intra-arterial fibrinolysis for acute transplant artery thrombosis. Transplantations had been performed 29 days to 10 years before the fibrinolysis. Fibrinolysis was carried out by using recombitant tissue plasminogen activator (n=1) or urokinase (n=3). In one patient, anuric for 13 hr at admittance, fibrinolysis could not revascularize the graft artery. In a second patient, anuric for 48 hr at admittance, fibrinolysis did revascularize the graft artery, but dialysis could not be discontinued. In the two remaining patients, anuric for 19 and 20 hr at admittance, the graft artery was successfully revascularized and dialysis could be discontinued 1 week later. One of these two patients returned to dialysis 71 months later because of chronic rejection. Thirty-four months after the acute episode, the remaining patient had a patent artery and did not require dialysis. CONCLUSIONS: Fibrinolysis seems an efficient treatment that may save transplants after up to 24 hr of the arterial occlusion. PMID- 11884936 TI - Recurrence of nephrotic syndrome in kidney grafts of patients with congenital nephrotic syndrome of the Finnish type: role of nephrin. AB - BACKGROUND: Congenital nephrotic syndrome of the Finnish type (CNF, NPHS1) is caused by mutations in the NPHS1 gene. NPHS1 codes for nephrin, a cell adhesion protein located at the glomerular slit diaphragm. Renal transplantation is the only treatment option for most patients with NPHS1. We have previously described recurrence of severe proteinuria in grafts transplanted to children with NPHS1. Here we studied the pathophysiology of this proteinuria. METHODS: Clinical data, light and electron microscopic findings as well as the expression of nephrin in the proteinuric grafts were studied. The patients' sera were screened for antibodies against kidney glomerulus and nephrin molecule using indirect immunofluorescence and ELISA. RESULTS: Fifteen episodes of recurrent nephrotic syndrome occurred in 13 (25%) of 51 grafts transplanted to 45 Finnish children with NPHS1. All nine patients with recurrence had a Fin-major/Fin-major genotype, which leads to absence of nephrin in the native kidney. Rescue therapy (cyclophosphamide) was successful in seven episodes, but six kidneys were lost due to this process. Antibodies reacting against glomerulus were found in eight, and high anti-nephrin antibody levels were detected in four of the nine patients. In electron microscopy, the fusion of the foot process and decreases in the detectable slit diaphragms in the podocyte pores were observed. The expression of nephrin mRNA was markedly reduced in two, and granular staining for nephrin was seen in three of five grafts. CONCLUSIONS: Circulating anti-nephrin antibodies seem to have a pathogenic role in the development of heavy proteinuria in kidney grafts of NPHS1 patients with Fin-major/Fin-major genotype. PMID- 11884938 TI - Kidney transplant in children weighing less than 15 kg: donor selection and technical considerations. AB - BACKGROUND: Small children represent a challenging patient group in kidney transplantation (KTx). The aim of this study was to analyze patient and donor data influencing outcome in children that weighed <15 kg. METHODS: Sixty-eight kidneys were transplanted in 64 children that weighed <15 kg. In 44 cases, kidneys came from cadaveric donors (CAD) and in 24 cases from living-related donors (LRD). Grafts were placed transperitoneally via midline incision (n=16) or extraperitoneally to the iliac fossa (n=52). Vascular anastomoses were routinely performed to the aorta and vena cava even when the extraperitoneal approach was used. RESULTS: Vascular thrombosis was observed in two (3%), urinary leaks in five (7%), and stenosis in four (6%) patients. In six children receiving organs from adults to the iliac fossa, wound closure was performed using an absorbable mesh to avoid organ compression. Initial graft function occurred in 60 cases (88%). Frequency of initial graft function was significantly higher after KTx from LRD (100%) compared with CAD (82%). The 1-, 5-, and 10-year patient survival was 93%, 91%, and 91%, respectively, and the 1-, 5-, and 10-year graft survival was 92%, 85%, and 85%, respectively. There was no significant difference in patient and graft survival when KTx from LRD and CAD were compared. Within the CAD group, graft survival was improved using kidneys from donors >12 years compared with younger donors. CONCLUSION: Despite size discrepancy between recipients and grafts, KTx is feasible in children that weigh <15 kg by using an improved surgical technique even when adult organs are placed to the iliac fossa. PMID- 11884939 TI - Arteroportal fistulas between the accessory right hepatic, gastroduodenal and superior mesenteric arteries and portal vein: a difficult technical problem to overcome in liver transplantation. AB - BACKGROUND: Fistulous communications between the accessory right hepatic (ARHA), gastroduodenal (GD), and superior mesenteric (SMA) arteries and the portal vein (PV) may represent a contraindication for liver transplantation (LT). MATERIAL: A patient with HCV-related liver cirrhosis and progressive liver decompensation underwent preoperative LT work-up. Doppler ultrasound (DU), Angiography and MRI revealed arteroportal fistulas (APF) and diversion of mesenteric-splenoportal flow through spontaneous splenorenal shunts (SSRS) in the systemic circulation. The patient was transplanted and the ARHA and GDA were distally sectioned; the HA was anastomosed to the donor HA; the superior mesenteric vein (SMV) was detached from the splenopancreatic venous bed by sectioning and ligating the Henle trunk, by ligating an posterior-inferior pancreatic vein and, finally, by positioning an iliac vein interposition graft between the SMV and the donor PV. The postanastomotic SMV trunk and recipient PV were ligated below and above the pancreatic head, respectively. RESULTS: Reperfusion and late liver function were good. DU and MRI studies showed an effective portal flow and the maintenance of a normal splenopancreatic vein outflow through the SSRS. DISCUSSION: APF represent a serious clinical problem, particularly in patients who need LT. The persistence of arterial flow into the PV is dangerous for the long-term liver function. A particular surgical strategy, strictly tailored to the hemodynamic conditions, has to be planned. CONCLUSIONS: Extrahepatic multiple APF would no longer to represent a contraindication to LT, although this claim needs to be confirmed in the light of further experience and a longer-term follow-up. PMID- 11884940 TI - Clinical and laboratory evaluation of the safety of a bioartificial liver assist device for potential transmission of porcine endogenous retrovirus. AB - BACKGROUND: The potential risk of transmission of porcine endogenous retroviruses (PERV) from xenogeneic donors into humans has been widely debated. Because we were involved in a phase I/II clinical trial using a bioartificial liver support system (BLSS), we proceeded to evaluate the biosafety of this device. MATERIALS AND METHODS: The system being evaluated contains primary porcine hepatocytes freshly isolated from pathogen-free, purpose-raised herd. Isolated hepatocytes were installed in the shell, which is separated by a semipermeable membrane (100 kD nominal cutoff) from the lumen through which the patients' whole blood is circulated. Both before and at defined intervals posthemoperfusion, patients' blood was obtained for screening. Additionally, effluent collected from a clinical bioreactor was analyzed. The presence of viral particles was estimated by reverse transcriptase-polymerase chain reaction (RT-PCR) and RT assays. For the detection of pig genomic and mitochondrial DNA, sequence-specific PCR (SS PCR) was used. Finally, the presence of infectious viral particles in the samples was ascertained by exposure to the PERV-susceptible human cell line HEK-293. RESULTS: PERV transcripts, RT activity, and infectious PERV particles were not detected in the luminal effluent of a bioreactor. Culture supernatant from untreated control or mitogen-treated porcine hepatocytes (cleared of cellular debris) also failed to infect HEK-293 cell lines. Finally, RT-PCR, SS-PCR, and PERV-specific RT assay detected no PERV infection in the blood samples obtained from five study patients both before and at various times post-hemoperfusion. CONCLUSION: Although longer patient follow-up is required and mandated to unequivocally establish the biosafety of this device and related bioartificial organ systems, these analyses support the conclusion that when used under standard operational conditions, the BLSS is safe. PMID- 11884941 TI - CD97-decay-accelerating factor interaction is not involved in leukocyte adhesion to endothelial cells. AB - BACKGROUND: Effective improvement in xenograft survival is achieved using transplants from transgenic pigs expressing human complement (C) regulatory proteins, including decay-accelerating factor (DAF), CD59, and CD46 on endothelial cells (ECs). The aim of this study was to investigate whether human DAF expression in porcine ECs, as well as regulating C activation, can modify intercellular events through its interaction with its receptor, CD97, on human leukocytes. METHODS: Cellular interactions between human leukocytes and porcine ECs were investigated in vitro using ECs from either wild-type or DAF-transgenic pigs. Static leukocyte adhesion and T cell activation assays were performed using porcine ECs as target or effector cells, respectively. The role of the DAF-CD97 interaction was investigated using specific blocking monoclonal antibodies (mAbs) against human DAF and its receptor, CD97, in adhesion assays. RESULTS: Adhesion of U937 or Jurkat T cells, both expressing human DAF and CD97, was quantitatively similar for wild-type and transgenic-DAF-expressing pig ECs. Furthermore, blocking the CD97-DAF interaction did not inhibit xenogeneic leukocyte endothelium adhesion, whereas blocking the very late antigen 4-vascular cell adhesion molecule-1 pathway reduced this adhesion by 50-80%. Furthermore, DAF and CD97 expression was not up-regulated during tumor necrosis factor-alpha- or lipopolysaccharide-mediated EC activation, unlike the adhesion molecules E selectin, vascular cell adhesion molecule-1, and intracellular adhesion molecule (ICAM)-1. CONCLUSION: We found that high levels of human DAF expressed on ECs abrogates C-mediated cell damage but did not affect the in vitro adhesive properties or antigen-presenting cell function of genetically modified porcine ECs. PMID- 11884942 TI - CD4+ T cells initiate pancreatic islet xenograft rejection via an interferon gamma-dependent recruitment of macrophages and natural killer cells. AB - BACKGROUND: In this study, the mechanisms by which CD4+ T cells interact with the innate immune system in xenograft rejection were investigated. METHODS: Fetal pig pancreas (FPP) grafts were transplanted into female SCID mice. The FPP recipient SCID mice were reconstituted with exogenous leukocytes obtained from male BALB/c mice. RESULTS: Although nonreconstituted SCID recipients or recipients reconstituted with CD4+ T cell-depleted leukocytes showed indefinite FPP graft survival with very few macrophages infiltrating their grafts, reconstitution of SCID recipients with as few as 2x10(5) CD4+ T cells was sufficient to induce rapid xenograft rejection. CD4+ T cells secreted interferon-gamma but not interleukin-4 and initiated the activation and accumulation of macrophages and natural killer cells, that were responsible for the rapid graft destruction. Suppression of interferon-gamma prolonged graft survival and suppressed the macrophages and natural killer cell accumulation and activation. CONCLUSIONS: These results demonstrate that CD4+ T cell-dependent cellular xenograft rejection was a result of macrophage and natural killer cell accumulation and activation, but was not mediated by eosinophils. Consistent with this was the finding that interferon-gamma but not interleukin-4 was in part responsible for mediating this effect. PMID- 11884943 TI - Modeling chronic lung allograft rejection in miniature swine. AB - BACKGROUND: The success of lung transplantation has been limited by the perplexing problem of chronic rejection. The development of a large-animal model for the systematic study of the mechanisms underlying chronic lung rejection has been problematic. We have developed a new preclinical model of chronic lung rejection using MHC-inbred miniature swine. METHODS: Using standard operative techniques, four orthotopic left lung allografts were performed using MHC matched, minor-antigen-mismatched donors. Recipient animals received a 12-day course of postoperative cyclosporine. Grafts were followed with open biopsies and high-resolution computed tomography. Cellular immune responses were monitored by mixed lymphocyte reaction, cytometric analysis of graft-infiltrating lymphocytes, and skin grafting. RESULTS: All grafts survived > or = 5 months and developed manifestations of chronic rejection, including obliterative bronchiolitis, interstitial fibrosis, and occlusive vasculopathy. A mononuclear infiltrate was also present in all grafts by the fourth posttransplant month. High-resolution computed tomography demonstrated several cardinal radiographic findings known to correlate with chronic rejection. Cytometric analysis of graft-infiltrating lymphocytes showed a predominance of CD8+ cells. The development of alloreactivity in the host was confirmed by mixed lymphocyte reaction and skin grafting. CONCLUSIONS: We report a reproducible, whole-lung, large-animal model of chronic lung rejection. In this immunogenetically defined construct, we have observed a full spectrum of histopathologic lesions that reproduce with fidelity those lesions observed in human lung transplant recipients suffering from chronic rejection. We anticipate that this preclinical model will facilitate further study of the pathogenesis and therapy of chronic lung rejection. PMID- 11884944 TI - Treatment with anti-CD154 antibody and donor-specific transfusion prevents acute rejection of myoblast transplantation. AB - BACKGROUND: Achieving immunological tolerance to transplanted myoblasts would reduce the adverse effects associated with the sustained immunosuppression required for this experimental therapeutic approach in Duchenne muscular dystrophic patients. METHODS: Mdx mice were transplanted with fully allogeneic BALB/c myoblasts in the tibialis anterior muscles. Seven days before transplantation (-7), host mice received 107 total donor spleen cells i.v. (donor specific transfusion, DST) with 500 microg of anti-CD154 mAb i.p. on days -7, -4, 0, +4. RESULTS: Results showed a high level of dystrophin expression in 83, 60, and 20% of the mice 1, 3, and 6 months, respectively, after transplantation of myoblasts. No antibodies against the donor cells were produced up to 3 months after transplantation. However, abundant activated cytotoxic cells were present in muscles still expressing high percentage of dystrophin positive fibers. CONCLUSIONS: In conclusion, the DST + anti-CD154 mAb treatments effectively prolonged myoblast survival, but this treatment could not develop tolerance to complete allogeneic myoblast transplantation. PMID- 11884945 TI - Cellular cardiomyoplasty in a transgenic mouse model. AB - BACKGROUND: Recent progress in the cardiotypic differentiation of embryonic and somatic stem cells opens novel prospects for the treatment of cardiovascular disorders. The aim of the present study was to develop a novel surgical approach that allows standardized cellular cardiomyoplasty in mouse with low-perioperative mortality. METHODS: Reproducible transmural lesions were generated by cryoinjury followed by intramural injection of embryonic cardiomyocytes using a newly designed holding device and vital dye staining. This approach was validated with a transgenic mouse model, in which the live reporter gene-enhanced green fluorescent protein (EGFP) is under control of a cardiac-specific promoter. RESULTS: The perioperative mortality was 10%. The engrafted EGFP-positive cardiomyocytes could be identified in a high percentage (72.2%, n=36) of operated animals. CONCLUSIONS: This novel approach enables reliable cellular replacement therapy in mouse and greatly facilitates the analysis of its molecular, cellular, and functional efficacy. PMID- 11884947 TI - Benefits of chronic plasmapheresis and intravenous heme-albumin in erythropoietic protoporphyria after orthotopic liver transplantation. AB - Erythropoietic protoporphyria (EPP) is characterized by a deficiency of ferrochelatase the final enzyme of the heme biosynthetic pathway. Patients with EPP may overproduce protoporphyrin IX, chiefly in developing erythrocytes. In some, protoporphyrin accumulates and causes toxicity, particularly to the skin and liver. Orthotopic liver transplantation (OLT) treats the severe liver disease that sometimes occurs in EPP; however, it does not correct the underlying metabolic disorder. We recently reported a patient with EPP who was improved with plasmapheresis and i.v. heme-albumin before OLT. Subsequently he developed histological and biochemical evidence of recurrent hepatotoxicity from protoporphyrin in the graft liver. We now report successful treatment of the patient with additional plasmapheresis and heme-albumin with improvement of hepatic histological and biochemical abnormalities. We conclude that plasmapheresis and heme-albumin are of benefit in EPP complicated by hepatotoxicity before and after liver transplantation. PMID- 11884946 TI - Epstein-Barr virus infection is associated with endothelial Bcl-2 expression in transplant liver allografts. AB - INTRODUCTION: In liver transplant recipients with Epstein-Barr virus (EBV) disease, we reported a low rate of acute rejection after stopping or markedly lowering immunosuppression. This observation led to the hypothesis that EBV, as a means of viral persistence, induces expression of antiapoptotic factors and these factors, in turn, confer protection to the transplanted organ. Bcl-2, an antiapoptotic factor induced by EBV in various host cells, is not normally expressed in the liver. We questioned whether bcl-2 is expressed in the transplanted liver and whether its expression is modified by EBV. MATERIALS AND METHODS: Retrospective liver biopsy specimen from liver transplant patients diagnosed with EBV (n=12) were examined for the presence of bcl-2 by immunohistochemistry and compared with EBV (-) transplant (n=15), and nontransplant (n=13) livers. RESULTS: The most significant finding was the presence of endothelial bcl-2 expression in the majority of EBV (+) transplant samples examined (67%) and its relative absence in the other two groups (P<0.005). There was also bcl-2 expression in the hepatocytes and lymphocytes of the majority of transplant liver samples, irrespective of EBV status. DISCUSSION: We have identified a strong association between EBV infection and endothelial bcl 2 expression in transplant livers. We also found that transplantation, in itself, was associated with bcl-2 expression in the hepatocytes and lymphocytes of liver allografts. PMID- 11884948 TI - Short course induction immunosuppression with thymoglobulin for renal transplant recipients. AB - BACKGROUND: The aim of this study was to demonstrate that 3-days of induction immunosuppression with thymoglobulin was as effective and safe as a 7-day course and reduced initial hospitalization after transplantation. METHODS: This was a prospective, nonrandomized trial of 40 consecutive patients receiving thymoglobulin induction for 3 days and followed for 1 year. An historical group of 48 patients that received 7 days of thymoglobulin served as controls. RESULTS: At 1 year, acute rejection (5 vs. 4%), graft survival (95 vs. 98%) and patient survival were similar; a composite end point of freedom from death, rejection, or graft loss, the event-free graft survival, was similar as was the safety profile. In the 3-day group, lymphocyte depletion was more sustained and initial hospitalization was significantly shorter (6 vs. 8 days). CONCLUSION: Three-day induction with thymoglobulin is as effective and safe as seven days, decreases initial hospitalization and causes more sustained lymphocyte depletion. PMID- 11884949 TI - Successful treatment of mucor infection after liver or pancreas-kidney transplantation. AB - BACKGROUND: Mucormycosis is a rare and opportunistic infection usually associated with hematologic diseases, diabetes mellitus, renal failure, solid tumors, and organ transplantation. METHODS: We present five cases of mucor infection after transplantation (three after a series of 750 orthotopic liver transplantation and two after a series of 13 simultaneous pancreas-kidney transplantation in patients with type 1 diabetes) subjected to medical and surgical treatment and analyze the factors related to the development of this infection. RESULTS: The clinical forms were two cutaneous (laparotomy wound or prior surgical drain site), two rhino maxillary, and one pulmonary. As risk factors for mucormycosis all patients had pre- or posttransplantation diabetes, and showed at least one episode of acute rejection that required aggressive immunosuppression (2-7 g of methylprednisolone; also three patients were treated with antithymocyte globulin [ATG] monoclonal antibody [orthoclone and/or OKT3]). We also found renal failure, acidosis, malnutrition, and Candida and cytomegalovirus infections as factors related to mucor infection. Diagnosis of fungal infection was confirmed by exudate or fluid culture in three cases and by biopsy in two. All patients were treated with liposomal amphotericin B (from 3.5 to 5.6 g of total dose) and resection until the surgical margins were free of infection. All patients survived after this severe infection. CONCLUSIONS: With an early diagnosis of mucormycosis by clinical findings, culture, or tissue biopsy, and aggressive treatment consisting of administration of liposomal amphotericin B and surgical resection of all infected tissue, excellent results are achieved. PMID- 11884951 TI - Isolated xeno and isoperfused rat liver: experimental procedure and results. PMID- 11884950 TI - Failure of reactivation of hepatitis B after liver transplantation in hepatitis B surface antigen-negative, core antibody-positive recipients. AB - BACKGROUND: Hepatitis B virus (HBV) infection after liver transplantation may occur in patients previously not exposed to the virus, but who receive an organ from a surface antigen (HBsAg)-negative, core antibody (HBcAb)-positive donor. The risk of HBV reactivation after liver transplantation in recipients that are HBsAg negative and HBcAb positive requires definition, because reactivation in kidney and bone marrow transplant patients has occurred. PATIENTS AND METHODS: A total of 409 HBsAg-negative patients underwent transplantation between April 1994 and June 1999. Pretransplantation sera were tested subsequently for HBcAb and HBsAb and posttransplantation sera for HBsAg. RESULTS: Of the 55 recipients who were positive for HBcAb, 48, who were immunosuppressed predominantly using tacrolimus, showed no evidence of HBV reactivation as shown by the absence of HBsAg (mean follow up 21.3+/-13.5 months). The remaining seven died within 6 months. CONCLUSIONS: In our experience, HBV reactivation did not occur in HBsAg negative, HBcAb-positive recipients after liver transplantation, most of whom were immunosuppressed with tacrolimus. We would not, therefore, currently recommend HBV prophylaxis in these patients. PMID- 11884952 TI - Patient acceptance and reliability of new Humulin/Humalog 3.0 ml prefilled insulin pen in ten Croatian diabetes centres. AB - BACKGROUND: The objective of this study was to assess feature preferences, patient acceptance, reliability, and safety of the new Lilly Humulin/Humalog 3.0 ml prefilled insulin pen in a clinical setting. MATERIAL/METHODS: A total of 330 patients in Croatia with type 1 or type 2 diabetes mellitus who required at least one injection of insulin per day were treated with Humulin 30/70 or Humalog for 4 to 6 weeks using the new prefilled pen. Questionnaires concerning various aspects of the pen performance were administered at endpoint. RESULTS: The features of the Lilly 3.0 ml pen device ranked most highly by patients (% of excellent or good ratings) were cartridge visibility (93%), attaching/replacing needles (93%), ease of dose correction (92%), checking insulin flow (90%) and dialling of insulin dose (89%). Features of the pen device rated most highly by patients in comparison with the delivery systems used before the study and the percentage of patients rating those features as much better or better were ease of dose correction (74%), cartridge visibility (67%), audible dialing clicks (55%) and size of dose numbers (52%). Most respondents (78%) preferred single-unit versus two-unit dosage increments. The majority of patients rated the new prefilled pen as being more convenient and easier to use, and indicated that it represented a significant or modest an improvement over their previous insulin injection method. CONCLUSIONS: The results of this study indicate that the new Lilly 3.0 ml prefilled pen is acceptable for patients who were previously using either reusable devices or the traditional syringe and vial. PMID- 11884953 TI - Postoperative pain control in cats: clinical trials with medetomidine and butorphanol. AB - OBJECTIVE: To evaluate the analgesic effects of medetomidine (MED) and butorphanol (BTO) in cats after ovariohysterectomy. STUDY DESIGN: A placebo controlled, blinded monocenter clinical study. ANIMALS: Healthy adult female client-owned cats. METHODS: Sixty-four cats weighing 3.15 +/- 0.6 kg, presented to the University of Helsinki's Small Animal Teaching Hospital for routine elective ovariohysterectomy, received MED at 15 microg/kg (n = 18), BTO at 0.1 mg/kg (n = 23), or saline (PL) (n = 23) intramuscularly immediately after ovariohysterectomy. Level of pain perception, degree of restlessness, and extent of sedation were scored subjectively before and at 30, 60, 90, and 120 minutes after test-drug administration. RESULTS: BTO provided the best pain relief, followed by MED. Saline provided the least pain relief. Both MED and BTO effectively and identically prevented postoperative restlessness. MED and BTO produced an identical degree of sedation that was better than the PL. CONCLUSIONS: Both MED (at 15 microg/kg) and BTO (at 0.1 mg/kg) prevent postoperative pain in cats after ovariohysterectomy. Clinical Relevance-MED and BTO are useful for preventing postoperative pain in cats. PMID- 11884955 TI - Nd:YAG laser-assisted modified Forssell's procedure for treatment of cribbing (crib-biting) in horses. AB - OBJECTIVE: To report an neodymium:yttrium-aluminum garnet (Nd:YAG) laser-assisted modified Forssell's surgical technique and outcome for treatment of cribbing (crib-biting) in horses. STUDY DESIGN: Retrospective clinical study. ANIMALS: Ten adult horses with stereotypic cribbing behavior. METHODS: Data were obtained from medical records and telephone conversations with owners, trainers, and veterinarians. Surgical technique involved an approximately 34-cm ventral median skin incision starting rostral to the larynx and extending caudally. A 10-cm section of the ventral branch of the spinal accessory nerve was removed, using an Nd:YAG laser at 25 W and continuous pulse with a contact, sculpted-fiber tip. After neurectomy, approximately 34-cm sections of the paired omohyoideus and sternothyrohyoideus muscles were removed starting 2 cm rostral to the ventral aspect of the larynx, at the basihyoid bone, using the Nd:YAG laser. RESULTS: Median horse age was 7 years (range, 1 to 11 years). Median surgical time was 90 minutes (range, 75 to 130 minutes). Long-term outcome (range, 7 to 72 months) was available for all horses. None of the horses had cribbing behavior after surgery, and all returned to their previous use. Four horses had complications (two of which were unrelated to the surgical site), but all recovered fully. CONCLUSION: The successful outcome we obtained is better than reported previously using a modified Forssell's technique. CLINICAL RELEVANCE: Surgical treatment for cribbing by Nd: YAG laser-assisted myectomy and neurectomy resulted in an excellent prognosis for resolution of the stereotypical behavior with minimal complications. PMID- 11884956 TI - Laparoscopic cryptorchidectomy using electrosurgical instrumentation in standing horses. AB - OBJECTIVE: To describe a technique for laparoscopic cryptorchidectomy in standing horses using electrosurgical instrumentation. STUDY DESIGN: Retrospective clinical study. ANIMALS OR SAMPLE POPULATION: Ten horses, 1 to 7 years of age, with unilaterally or bilaterally retained testes. METHODS: Food was withheld for a minimum of 12 to 24 hours. Horses were sedated using xylazine hydrochloride (0.5 to 1 mg/kg) and butorphanol tartrate (0.02 mg/kg) or detomidine hydrochloride (0.02 to 0.03 mg/kg) and restrained in standing stocks. Three portal sites in the paralumbar fossae were locally desensitized using 2% mepivacaine. After trocar and laparoscope insertion, the ipsilateral testicle, mesorchium, and ductus deferens were identified. The cranial mesorchium was coagulated with either monopolar (one horse) or bipolar (nine horses) electrosurgical forceps, and then the mesorchium, ductus deferens, and ligament of the tail of the epididymis were transected from cranial to caudal using laparoscopic scissors. Once the testis was freed, the transected mesorchium was inspected for hemorrhage and the testis was removed by connecting the two instrument portals (eight horses). In two horses, the testis was placed within a laparoscopic retrieval bag and then removed without enlarging the portal incision. If the testes were retained bilaterally, the retained contralateral testis was removed similarly through the opposite paralumbar fossa. If the contralateral testis was descended, it was removed by a standard, standing castration technique. RESULTS: Vessels of the mesorchium were adequately coagulated using bipolar and monopolar electrosurgical forceps. No immediate or short-term complications occurred in 10 horses at 3 to 11 months after surgery. CONCLUSION: Standing laparoscopic cryptorchidectomy can be performed easily and safely using electrosurgical instrumentation as the sole means of providing hemostasis of the equine mesorchium. CLINICAL RELEVANCE: Standing laparoscopic cryptorchidectomy using electrosurgical instrumentation provides a safe, reliable, and efficient alternative to achieve hemostasis of the equine mesorchium. PMID- 11884954 TI - A comparison of the mechanical strength of two stapled anastomosis techniques for equine small intestine. AB - OBJECTIVE: To compare bursting strength, time of construction, and cost of a closed one-stage, stapled functional end-to-end jejunojejunostomy (FEE) with a stapled side-to-side jejunojejunostomy (STS). STUDY DESIGN: Experimental, randomized block design. ANIMALS: Seven adult horses without gastrointestinal disease. METHODS: The jejunum was isolated, and three FEE, three STS, and three control segments were created in each horse using a randomized block design. Anastomosis time was recorded. The intraluminal pressure at failure and mode of failure were recorded. Length at failure was measured on digitized images. Bursting pressure (BP), bursting wall tension (BWT), anastomosis time, and cost were compared. RESULTS: Control jejunal segments were stronger (P < or = .0001) in bursting strength and bursting wall tension (P < or = .0001) than either anastomosis type; no difference was found between anastomosis types for either variable. Functional end-to-end jejunojejunostomy was significantly quicker and less costly than STS (P < or = .0001). CONCLUSIONS: Mechanically there were no significant differences between the FEE and STS techniques. The FEE technique maintained the physiologic direction of peristalsis of the segments, required less tissue manipulation, and was faster and more economical to create. CLINICAL RELEVANCE: The FEE is a clinically viable technique. PMID- 11884958 TI - Effect of a supplemental plate on the stiffness of a type I external fixator. AB - OBJECTIVE: To assess the effect of a supplemental plate on the stiffness of a six pin unilateral external skeletal fixator. STUDY DESIGN: Mechanical testing performed on models. METHODS: Wooden (birch) dowels were used to create five models of a fracture. A commercially available external fixation system was applied to the model with a uniform unilateral six-pin fixator design. The models were mechanically tested with and without a supplemental plate attached to the 2 clamps adjacent to the fracture gap. Testing was conducted in axial loading, medial to lateral bending, and cranial to caudal bending. RESULTS: Results showed a 4.42-fold increase in stiffness in axial load, a 4.23-fold increase in stiffness in medial to lateral bending, and a 1.94-fold increase in stiffness in cranial to caudal bending with the addition of the plate. CONCLUSIONS: The addition of a supplemental plate increases the mechanical stiffness of unilateral fixators. This was especially true in axial load and medial to lateral bending. CLINICAL RELEVANCE: A supplemental plate can be used with unilateral fixators to increase stiffness of the fixator. Conversely, the plate can be removed to decrease stiffness without the removal of fixation pins. PMID- 11884957 TI - Sensitivity of radiographic evaluation of radio-ulnar incongruence in the dog in vitro. AB - OBJECTIVE: To determine the sensitivity and specificity of radiographic evaluation of radio-ulnar incongruence in canine elbow joints in vitro. STUDY DESIGN: Radiographic evaluation of induced radio-ulnar incongruence in canine cadaveric forelimbs by radiologists blinded to study design. SAMPLE POPULATION: Six cadaveric canine left forelimbs. METHODS: Extended lateral, 90 degrees flexed lateral, and cranio-caudal projections were taken of the elbow joint of six cadaveric canine forelimbs. A four-pin, type I external skeletal fixator (ESF) with a linear motor side bar was attached to the medial aspect of the radius, and a 2-cm segment of bone was removed from the mid-diaphysis. A 3.5-mm cortical bone screw placed from the medial to lateral styloid processes prevented relative movement between the distal radius and ulna during radial shortening. The ESF was used to progressively shorten the radius in increments of 0.5 mm to a total of 4 mm. The three radiographic projections were repeated after each incremental change of length. After the study, each elbow joint was disarticulated to confirm the presence of a step defect. The original radiographs and three copies were randomized and then evaluated by four radiologists blinded to the study design. Radiologists were asked to evaluate whether the joint was normal or abnormal and if there was evidence of radio-ulnar incongruence. The ability of each radiologist to correctly identify congruent elbows (specificity) and incongruent elbows (sensitivity) was calculated. RESULTS: The median specificity was 86% using the lateral projection and 82% using the cranio-caudal projection. The median sensitivities using the lateral and cranio-caudal radiographic projections were 78% and 79%, respectively. The degree of radial shortening required for individual radiologists to achieve a sensitivity of 90% ranged from 1.5 mm to greater than 4 mm. CLINICAL RELEVANCE: Standard radiographic evaluation for radio ulnar incongruence in the dog may be associated with relatively poor sensitivity and specificity. Invasive surgical procedures and screening programs that rely on radiographic diagnosis of radio-ulnar incongruence should be discouraged until a more reliable method of diagnosis of this type of elbow joint incongruence is available. PMID- 11884960 TI - Effect of tibial plateau leveling on stability of the canine cranial cruciate deficient stifle joint: an in vitro study. AB - OBJECTIVE: To evaluate the effect of tibial plateau leveling on joint motion in canine stifle joints in which the cranial cruciate ligament (CCL) had been severed. STUDY DESIGN: In vitro cadaver study. ANIMALS: Six canine cadaver hind legs. METHODS: Radiographs of the stifle joints were made to evaluate the tibial plateau angle with respect to the long axis of the tibia. The specimens were mounted in a custom-made testing device to measure cranio-caudal translation of the tibia with respect to the femur. An axial load was applied to the tibia, and its position was recorded in the normal stifle, after transection of the CCL, and after tibial plateau leveling. Further, the amount of caudal tibial thrust was measured in the tibial plateau leveled specimen while series of eight linearly increasing axial tibial loads were applied. RESULTS: Transection of the CCL resulted in cranial tibial translation when axial tibial load was applied. After tibial plateau leveling, axial loading resulted in caudal translation of the tibia. Increasing axial tibial load caused a linear increase in caudal tibial thrust in all tibial plateau-leveled specimens. CONCLUSIONS: After tibial plateau leveling, axial tibial load generates caudal tibial thrust, which increases if additional axial load is applied. CLINICAL RELEVANCE: Tibial plateau leveling osteotomy may prevent cranial translation during weight bearing in dogs with CCL rupture by converting axial load into caudal tibial thrust. The amount of caudal tibial thrust seems to be proportional to the amount of weight bearing. PMID- 11884959 TI - Thoracoscopic visualization and ligation of the thoracic duct in dogs. AB - OBJECTIVE: To develop a technique for thoracoscopic visualization and ligation of the thoracic duct in dogs. STUDY DESIGN: In vivo experimental study. ANIMALS: Five mature, healthy dogs. METHODS: Dogs were normal based on physical examination, negative occult heartworm test, normal complete blood count and biochemical profile, and normal thoracic radiographs. The dogs were anesthetized, and a ventral midline laparotomy was performed for catheterization of a mesenteric lymphatic. Lymphangiography was performed to determine thoracic duct anatomy. Thoracoscopy was performed in the caudal, right hemithorax after single lung intubation or bronchial blockade. At least two 10-mm clips were placed across the thoracic duct in each dog. Lymphangiography was repeated to assess duct ligation. If complete duct occlusion was not achieved, thoracoscopy was repeated for additional clip placement. After surgery the dogs were euthanatized, and necropsies were performed. RESULTS: Lymphangiography showed that multiple branches of the thoracic duct were present in every dog; bilateral thoracic duct branches were most common. Thoracoscopic identification and ligation of the thoracic duct was successful in all five dogs. Two dogs required a second thoracoscopic procedure to completely occlude flow of contrast through the thoracic duct. Surgery time for thoracoscopy averaged 59 plus minus 9.6 minutes. Retroperitoneal contrast accumulation after thoracic duct ligation occurred in two dogs. One dog required bilateral pulmonary ventilation. CONCLUSION: Thoracoscopy can be used to visualize the thoracic duct for ligation in normal dogs. CLINICAL RELEVANCE: Thoracoscopic ligation of the thoracic duct may be a therapeutic option for management of chylothorax in dogs. PMID- 11884961 TI - An evaluation of two autologous tendon grafting techniques in ponies. AB - OBJECTIVE: To compare the healing and mechanical strength of a multiple split autologous tendon graft (MG) to a whole autologous tendon graft (WG) in the deep digital flexor tendon of ponies. STUDY DESIGN: In vitro evaluation of two different tendon-grafting techniques. ANIMALS OR SAMPLE POPULATION: Six ponies of mixed gender and age. METHODS: Tenotomies performed in forelimb deep digital flexor tendons (DDFT) distal to the insertion of the accessory ligament (AL-DDFT) were repaired with free autologous grafts from the hindlimb lateral digital extensor tendon (LDET). Grafts were either whole (WG) or split into three longitudinal strips (multiple graft, MG). Tendons and graft sites were collected and loaded (2.54 cm/s) to failure at either 4 or 8 weeks after surgery. Cross sectional area was determined by both impression cast (IC) and an inkblot (IB) method. Tissue maturity and inflammation were evaluated by microscopy. RESULTS: Gap formation was a consistent finding in all repair sites. No statistical differences were found in healing or mechanical variables between MG and WG techniques. The failure stress for the 8-week repairs (15.51 +/- 3.1 MPa IB and 11.73 +/- .77 MPa IC, 16.13 +/- 2.2 MPa IB and 10.22 +/- .76 MPa IC for MG and WG, respectively) were significantly greater (P <.0005) than for 4-week repairs (3.71 +/- 1.7 MPa IB and 2.68 +/- 1.44 MPa IC, 2.81 +/- 1.46 MPa IB and 2.3 +/- 1.7 MPa IC for MG and WG, respectively). The repair tissue was more mature (P <.05) at 8 weeks than at 4 weeks, but there was no significant difference in inflammatory responses at 4 and 8 weeks. CONCLUSIONS: There was a sixfold increase in strength between 4 and 8 weeks of healing, but no significant difference in healing or strength between the MG and WG techniques. CLINICAL RELEVANCE: In ponies, autologous tendon grafting contributes to a strong repair during the early convalescent period, but splitting a tendon graft seemingly offers no appreciable advantage over use of a whole graft. PMID- 11884962 TI - The effects of acetabular cup temperature and duration of cement pressurization on cement porosity in a canine total hip replacement model. AB - OBJECTIVE: To determine the effect of acetabular cup temperature and duration of cement pressurization on porosity of the acetabular cement mantel. STUDY DESIGN: In vitro study. METHODS: Twenty-four polyurethane foam blocks prepared for acetabular prosthetic implantation were implanted with polyethylene acetabular cups using four combinations and variations of temperature and pressure: (1) high temperature/short-term pressurization; (2) high temperature/long-term pressurization; (3) low temperature/long-term pressurization; and (4) low temperature short-term pressurization. Five 1-mm-thick slices were taken from the center of each block using a tissue processing system. The slices were scanned into a personal computer using a photo slide scanner. Imaging software was used to determine cement surface area and size, number, and distribution of pores. The quality of the cement-implant interface was subjectively evaluated. Statistical analysis of relative cement porosity was performed by a Kruskal-Wallis analysis of variance comparing the four groups individually and combining the short-term pressurization groups versus the long-term pressurization groups. RESULTS: There were no significant differences in cement porosity between the four test groups (P =.11). There were no significant differences in porosity between the combined groups (P =.48). CONCLUSIONS: There is no benefit in prewarming acetabular cups before implantation. There are no deleterious effects of short-term pressurization of the cement during implantation. PMID- 11884963 TI - Open peritoneal drainage versus primary closure for the treatment of septic peritonitis in dogs and cats: 42 cases (1993-1999). AB - OBJECTIVE: To determine survival rates in dogs and cats with septic peritonitis treated with open peritoneal drainage (OPD) versus primary closure (PC) after laparotomy. STUDY DESIGN: Retrospective analysis of medical records from Colorado State University Veterinary Teaching Hospital from 1993 to 1999. SAMPLE POPULATION: Thirty-six dogs and 6 cats with septic peritonitis documented by cytological examination or microbiological culture of abdominal fluid. METHODS: Medical records of dogs and cats with septic peritonitis treated by OPD or PC were reviewed. Age, weight, species, white blood cell (WBC) count, band neutrophil count, platelet count, serum glucose concentration, heart rate, body temperature, duration of hospitalization, and clinical outcome were recorded for each animal. Differences in treatments administered between the OPD and PC groups as well as the underlying cause of septic peritonitis were determined. RESULTS: There was no significant difference in survival between animals in the OPD versus PC groups (P =.26) with an overall survival rate of 71%. White blood cell count, band neutrophil count, platelet count, serum glucose and total bilirubin concentrations, heart rate, age, and weight were not significantly different between groups (P >.05). A significantly greater number of animals in the OPD group received plasma (P =.009), blood (P =.037), and a jejunostomy tube (P =.02) than animals in the PC group. There was a significant difference in the number of days spent in critical care unit with a mean of 6.0 +/- 4.1 days for the OPD group and 3.5 +/- 2.3 days for the PC group (P =.02). CONCLUSIONS: Open peritoneal drainage for the management of septic peritonitis in dogs and cats is an acceptable alternative to PC. PMID- 11884965 TI - Abdominal aortic aneurysms: basic mechanisms and clinical implications. PMID- 11884964 TI - External fixation of the lumbar spine in a canine model. AB - OBJECTIVE: To compare the mechanical properties of two types of external skeletal fixation of the lumbar spine with polymethylmethacrylate (PMMA)/Steinmann pin fixation in a canine unstable spine model. STUDY DESIGN: Cadaver study. SAMPLE POPULATION: Lumbar spines of 17 mature large-breed dogs. METHODS: Spine stiffness (N-m/deg) in flexion, extension, and rotation under physiological loading conditions and spine strength (N-m) in flexion were determined. Spines were destabilized at L3-L4, instrumented and retested. Fixation techniques included four-pin PMMA (PMMA4), eight-pin PMMA (PMMA8), eight-pin biplanar type I external skeletal fixator (ESF) (SK), and eight-pin spinal arch ESF (ARCHES). RESULTS: All fixation groups were as stiff as intact spines in extension and rotation and were significantly stiffer in flexion. In flexion, both PMMA8 and ARCHES were significantly stiffer than SK, and PMMA8 was significantly stiffer than PMMA4. In rotation, PMMA8 and ARCHES were significantly stiffer than SK, and in flexion to failure, PMMA8 and ARCHES were significantly stiffer than PMMA4. CONCLUSIONS: External skeletal spinal fixation (ESSF) has mechanical properties comparable to more commonly used PMMA/pin internal fixation techniques. CLINICAL RELEVANCE: External fixation of the canine spine has several potential advantages over internal fixation including minimal dissection for pin placement, the ability to span affected vertebrae with placement of implants distant from the site of injury, postoperative adjustability, and complete removal of implants after healing. This study supports the biomechanical stability ESSF of the canine lumbar spine. Further studies are indicated to evaluate zones of consistently safe and secure placement of pins and clinical efficacy. PMID- 11884967 TI - Investigations into the safety and immunogenicity of a killed oral cholera vaccine developed in Viet Nam. AB - OBJECTIVE: To evaluate a killed oral cholera vaccine produced in Viet Nam, and to compare the Vietnamese vaccine with one that is licensed internationally. METHOD: Two-dose regimens of a locally produced, bivalent, anti-O1, anti-O139 killed oral whole-cell cholera vaccine (biv-WC) and of a commercially available, monovalent (anti-O1) oral recombinant B subunit-killed whole-cell cholera vaccine (rBS-WC) were compared in two trials in Viet Nam. In the first trial, 144 adults were randomized to biv-WC with or without buffer, rBS-WC with buffer, or placebo without buffer. In the second, 103 children aged 1-12 years were randomized to biv-WC without buffer, rBS-WC with buffer, or placebo without buffer. FINDINGS: No regimen was associated with significant side-effects. In adults, ca 60% of recipients of either vaccine exhibited at least fourfold serum anti-O1 vibriocidal antibody responses and ca 40% of recipients of biv-WC demonstrated anti-O139 vibriocidal responses. Both anti-O1 (ca 90% in each vaccine groupand anti-O139 (68% in the biv-WC group) vibriocidal responses occurred more frequently in children. The responses to biv-WC were unaffected by the receipt of buffer. CONCLUSION: It was concluded that biv-WC was safe and immunogenic, that it could be administered without buffer, and that it could elicit robust immune responses even in children, for whom the risk of endemic cholera is highest. PMID- 11884968 TI - Death rate variation in US subpopulations. AB - OBJECTIVE: To account for variations in death rates in population subgroups of the USA. METHODS: Factors associated with age-adjusted death rates in 366 metropolitan and non- metropolitan areas of the United States were examined for 1990-92. The rates ranged from 690 to 1108 per 100 000 population (mean = 885 +/- 78 per 100 000). FINDINGS: Least squares regression analysis explained 71% of this variance. Factors with the strongest independent positive association were ethnicity (African-American), less than a high school education, high Medicare expenditures, and location in western or southern regions. Factors with the strongest independent negative associations were employment in agriculture and forestry, ethnicity (Hispanic) and per capita income. CONCLUSION: Additional research at the individual level is needed to determine if these associations are causal, since some of the factors with the strongest associations, such as education, have long latency periods. PMID- 11884969 TI - Etiology of child mortality in Goroka, Papua New Guinea: a prospective two-year study. AB - OBJECTIVE: To collect accurate data on disease- and microbial-specific causes and avoidable factors in child deaths in a developing country. METHODS: A systematic prospective audit of deaths of children seen at Goroka Hospital in the highlands of Papua New Guinea was carried out. Over a 24-month period, we studied 353 consecutive deaths of children: 126 neonates, 186 children aged 1-59 months, and 41 children aged 5-12 years. FINDINGS: The most frequent age-specific clinical diagnoses were as follows: for neonates--very low birth weight, septicaemia, birth asphyxia and congenital syphilis; for children aged 1-59 months--pneumonia, septicaemia, marasmus and meningitis; and for children aged 5-12 years- malignancies and septicaemia. At least one microbial cause of death was identified for 179 (50.7%) children and two or more were identified for 37 (10.5%). Nine microbial pathogens accounted for 41% of all childhood deaths and 76% of all deaths that had any infective component. Potentially avoidable factors were identified for 177 (50%) of deaths. The most frequently occurring factors were as follows: no antenatal care in high-risk pregnancies (8.8% of all deaths), very delayed presentation (7.9%), vaccine-preventable diseases (7.9%), informal adoption or child abandonment leading to severe malnutrition (5.7%), and lack of screening for maternal syphilis (5.4%). Sepsis due to enteric Gram-negative bacilli occurred in 87 (24.6%). The strongest associations with death from Gram- negative sepsis were adoption/abandonment leading to severe malnutrition, village births, and prolonged hospital stay. CONCLUSIONS: Reductions in child mortality will depend on addressing the commonest causes of death, which include disease states, microbial pathogens, adverse social circumstances and health service failures. Systematic mortality audits in selected regions where child mortality is high may be useful for setting priorities, estimating the potential benefit of specific and non-specific interventions, and providing continuous feedback on the quality of care provided and the outcome of health reforms. PMID- 11884970 TI - Comparison of cohort smoking intensities in Denmark and the Netherlands. AB - OBJECTIVE: To assess the usefulness of the general framework of the smoking epidemic. METHODS: We use lung cancer mortality as an indicator for smoking intensity and employ an age-cohort model to accommodate the long-lasting and cumulative effects. RESULTS: Dutch males have higher risks than Danish males, but the risks for the younger cohorts have been declining faster in the Netherlands than in Denmark. Danish women have about twice the risk of Dutch women, and in both countries the risks for the younger cohorts are increasing. The smoking epidemic began at about the same time in Denmark and the Netherlands. Dutch males, however, seem to have smoked more but to have given up smoking more quickly than Danish males. Danish females were quicker to take up smoking than Dutch females. CONCLUSIONS: Within the general framework of the smoking epidemic, differences in timing and levels can produce large differences between countries. For the purposes of assessing smoking-related risks, including projections, the smoking epidemic framework therefore has to be tailored to each study population. PMID- 11884971 TI - Monitoring one-year compliance to antihypertension medication in the Seychelles. AB - OBJECTIVE: To examine the compliance to medication among newly diagnosed hypertensive patients screened from the general population of the Seychelles, a rapidly developing country. METHODS: Among the 1067 participants to a population based survey for cardiovascular risk factors, hypertension was discovered in 50 (previously unaware of having hypertension and having blood pressure > or = 160/95 mmHg over 3 visits). These 50 patients were placed on a daily one-pill regimen of medication (bendrofluazide, atenolol, or a combination of hydrochlorothiazide and atenolol) and compliance to the regimen was assessed over 12 months using electronic pill containers. Satisfactory compliance was defined as taking the medication on 6 or 7 days a week on average (which corresponds to a mean compliance level of > or = 86%). FINDINGS: In the first month, fewer than half (46%) of the new hypertension patients achieved satisfactory compliance, and only about one-quarter (26%) achieved this level by the twelfth month. Compliance was better among the 23 participants who regularly attended medical follow-up, with nearly three-quarters of these patients (74%) achieving satisfactory compliance during the first month and over one-half (55%) by the twelfth month. There was a direct association between mean 12-month compliance level and having a highly skilled occupation; having good health awareness; and regularly attending medical appointments. In contrast, there was an inverse relationship between mean compliance level and heavy drinking. CONCLUSION: The low proportion of people selected from the general population who were capable of sustaining satisfactory compliance to antihypertension medication may correspond to the maximum effectiveness of medication interventions based on a screening and treatment strategy in the general population. The results stress the need for both high-risk and population approaches to improve hypertension control. PMID- 11884972 TI - The risk of Ascaris lumbricoides infection in children as an environmental health indicator to guide preventive activities in Caparao and Alto Caparao, Brazil. AB - OBJECTIVE: To develop an environmental health indicator for use as a basis for developing preventive measures against Ascaris lumbricoides infection in children from the rural municipalities of Caparao and Alto Caparao, in Minas Gerais, Brazil. METHODS: A cross-sectional survey was conducted between May and September 1998 among 1171 children under 14 years of age living in 588 dwellings selected from 11 communities. Trained interviewers used a questionnaire to identify risk factors for infection (socioeconomic, sanitation and hygiene variables) and collected stool samples from each child for parasitological tests. RESULTS: The overall prevalence of A. lumbricoides infection was 12.2%. The results showed the protective effects of availability of water in the washbasin and better hygiene, sanitation and socioeconomic status; the interactive effect of crowding was five times larger in households without water in the washbasin than in those having water. There was a statistically significant association between infection and children's age. CONCLUSION: The environmental health indicator, which incorporated the most significant biological, environmental and social factors associated with the risk of A. lumbricoides infection in children from these communities, should contribute to the development of surveillance tools and health protection measures in this population. PMID- 11884973 TI - An economic analysis of midwifery training programmes in South Kalimantan, Indonesia. AB - In order to improve the knowledge and skills of midwives at health facilities and those based in villages in South Kalimantan, Indonesia, three in-service training programmes were carried out during 1995-98. A scheme used for both facility and village midwives included training at training centres, peer review and continuing education. One restricted to village midwives involved an internship programme in district hospitals. The incremental cost-effectiveness of these programmes was assessed from the standpoint of the health care provider. It was estimated that the first scheme could be expanded to increase the number of competent midwives based in facilities and villages in South Kalimantan by 1% at incremental costs of US$ 764.6 and US$ 1175.7 respectively, and that replication beyond South Kalimantan could increase the number of competent midwives based in facilities and villages by 1% at incremental costs of US$ 1225.5 and US$ 1786.4 per midwife respectively. It was also estimated that the number of competent village midwives could be increased by 1% at an incremental cost of US$ 898.1 per intern if replicated elsewhere, and at a cost of US$ 146.2 per intern for expanding the scheme in South Kalimantan. It was not clear whether the training programmes were more or less cost-effective than other safe motherhood interventions because the nature of the outcome measures hindered comparison. PMID- 11884974 TI - The implications of health sector reform for human resources development. AB - The authors argue that "health for all" is not achievable in most countries without health sector reform that incorporates a process of coordinated health and human resources development. They examine the situation in countries in the Eastern Mediterranean Region of the World Health Organization. Though advances have been made, further progress is inhibited by the limited adaptation of traditional health service structures and processes in many of these countries. National reform strategies are needed. These require the active participation of health professional associations and academic training institutions as well as health service managers. The paper indicates some of the initiatives required and suggests that the starting point for many countries should be a rigorous appraisal of the current state of human resources development in health. PMID- 11884976 TI - Drawers of water: domestic water use in East Africa. 1972. PMID- 11884977 TI - Water, sanitation, and hygiene evaluation issues. PMID- 11884978 TI - Impact of the Bosnian conflict on the health of women and children. PMID- 11884979 TI - The role of RT-PCR assay of oral fluid for diagnosis and surveillance of measles, mumps and rubella. PMID- 11884980 TI - HIV/AIDS surges in Eastern Europe--Asia-Pacific next? PMID- 11884981 TI - Haemoglobin variant gives strong protection against malaria. PMID- 11884982 TI - Anti-inflammatory drugs slash Alzheimer risk. PMID- 11884983 TI - WHO attacks tobacco sponsorship of sports. PMID- 11884985 TI - Lead, unsafe at any level. PMID- 11884986 TI - [Pneumology 2002]. PMID- 11884987 TI - [Antimycotic treatment of pulmonary aspergilloma in patients without neutropenia]. AB - BACKGROUND AND OBJECTIVE: Pulmonary aspergillomas are associated with a high morbidity and mortality. There are only very few data about non-surgical treatment in immunocompetent patients. METHODS: We evaluated 30 patients (19 male/11 female; mean age: 54 years) with pulmonary aspergillosis, their symptoms, treatment and outcome during a time period of 9 years. All patients had either definitive (18/60 %) or probable (12/40 %) aspergillosis with a cultural confirmation. RESULTS: Underlying diseases were tuberculosis (16/53 %), malignancy (8/30 %), COPD (15/50 %), pneumonia (3/10 %). Only five patients were immunocompromised (steroid medication: n = 4, chronic lymphatic leukemia: n = 1) All patients had a contraindication against surgery, 26 received antifungal treatment (mostly with voriconazole or itraconazole). The overall response was 61 %, there was an improvement of radiological signs in seven (23 %). Especially hemoptysis and dyspnea resolved. 12 patients died during the 9 years (40 %), nine from their underlying disease, three from the pulmonary aspergilloses (hemoptysis: n = 2, secondary invasive aspergillosis: n = 1). CONCLUSION: Pulmonary aspergilloma in its chronic form is a disease of patients with pulmonary disease but who are immunocompetent. Antifungal treatment seems to be a therapeutic option, if surgery is not possible. PMID- 11884989 TI - [Turner syndrome, autoimmune thyroiditis and partial "empty sella turcica."--an unusual case in a progressed aging unrecognized diagnostic combination]. PMID- 11884988 TI - [Undetected tracheal tumours responsible for ventilator-dependency]. AB - HISTORY AND CLINICAL FINDINGS: Case 1 A patient with former carcinoma of the larynx became dependent on mechanical ventilation. She failed to be weaned from the respirator because of severe bronchial obstruction, therefore she was transferred to a weaning center. Case 2 A COPD patient with respirator dependency due to infectious exacerbation underwent percutaneous tracheostomy shortly after primary intubation. Status asthmaticus was considered to be the reason of following unsuccessful weaning. INVESTIGATIONS, DIAGNOSIS AND TREATMENT: Case 1 Performing a bronchoscopy the diagnosis of a central tumor (local recurrence) was found causing nearly total obstruction of the trachea. The ensuing treatment was restricted to palliation. Case 2 After transferral to the weaning center a small cell lung cancer located in the central tracheal was identified by bronchoscopy. The tumor masses were exstirpated by laser technique and the patient was weaned immediately afterwards. Chemotherapy and radiation of the mediastinum were performed. CONCLUSIONS: Fibreoptic bronchoscopy is an essential tool concerning diagnosis and treatment of tracheal tumors which may cause difficult weaning from mechanical ventilation. Every percutaneous tracheostomy should be performed with endoscopical guidance. PMID- 11884990 TI - [Genetic diagnosis and consultation in bronchiectasia. First diagnosis of cystic fibrosis in an adult]. PMID- 11884991 TI - [Chronic obstructive pulmonary disease: pathophysiology and diagnosis]. PMID- 11884992 TI - [Chronic obstructive pulmonary disease: treatment]. PMID- 11884996 TI - [Interstitial pulmonary disease in systemic scleroderma: diagnosis]. PMID- 11884997 TI - [Interstitial pulmonary disease in systemic scleroderma: treatment]. PMID- 11884998 TI - [Singing in asthma--does budesonide harm the voice box?]. PMID- 11884999 TI - [Diagnosis of pulmonary artery embolism.. Part 1]. PMID- 11885000 TI - [Diagnosis of pulmonary artery embolism. Reply]. PMID- 11885001 TI - [Interventional radiology - quo vadis?]. PMID- 11885002 TI - [Multislice CT of the abdomen--current indications and future trends]. AB - Multislice CT systems allow the simultaneous acquisition of multiple slices per gantry rotation. In combination with faster gantry rotation times of 0.5 seconds, the abdominal structures can be displayed in higher spatial and temporal resolution. In MS-CT of the liver it is possible to scan the entire organ with an optimal slice thickness of 2 - 3 mm within a defined perfusion phase in less than 10 seconds. This results in an improved detection and characterization of focal liver lesions. A high-quality, 3-dimensional reconstruction of the hepatic arterial and portalvenous system is obtained with the same data set. The diagnostic use of the simultaneously acquired perfusion data will lead to a better characterization of focal liver lesions in the future. The diagnostics of the pancreas also profits from MS-CT, especially for the detection of small tumors and the evaluation of resectability of a pancreatic carcinoma. All abdominal structures can be displayed in a coronal view without loss of image quality because of the almost isotropic voxels obtained. This proves to be beneficial for the preoperative diagnostics of renal cell carcinomas, especially if the tumor extension into adjacent organs (e. g., liver or spleen) in the longitudinal direction has to be evaluated. The multiplanar display and the sophisticated 3-dimensional reconstruction tools have a substantial value for the abdominal CT angiography. It proves to have a major diagnostic impact on acute abdominal aortic and visceral arterial diseases because even large distances in the z-direction can be covered with high spatial resolution. PMID- 11885003 TI - Real-time-MR guidance for placement of a self-made fully MR-compatible atrial septal occluder: in vitro test. AB - PURPOSE: This in vitro study investigated the feasibility to visualize the placement of three different atrial septal occluder systems using real-time MR control. METHODS: The experiments were performed on an interventional 1.5 T high field whole body system. Real-time MR imaging was achieved by radial or spiral k space filling in conjunction with the sliding window reconstruction technique yielding an imaging speed of 15 frames per second. The CardioSeal, Amplatzer Septal Occluder and a specially designed MR-compatible closure device were tested in a water bath. A punctured plastic wall served as model for the atrial septal defect. RESULTS: The delivery systems of the CardioSeal and Amplatzer Occluder were ferromagnetic and caused substantial artifacts, making the device placement impossible, even if the magnetic forces would have been acceptable. The self-made prototype caused only minor susceptibility artifacts allowing its visualization on the MR images. The MR imaging techniques applied enabled real-time control of the occluder including steering through the artificial septal foramen and visualization of the occluder deployment. CONCLUSION: Real-time MR imaging allows for guidance and placement of an MR-compatible septal occluder in vitro suggesting the feasibility to perform atrial septal occlusion under MR-guidance in vivo as well. PMID- 11885004 TI - [Solid-pseudopapillary tumors in childhood]. AB - We report on fife female patients with solid pseudopapillary tumors of the pancreas. The tumors are extremely rare in children. They occur mainly in adolescent and young adult females. The tumors are neoplasms of low malignancy with infrequent metastases, for instance, in the liver or the peritoneum. Although the tumors had reached a large diameter, all of them underwent complete tumor resection. After that the patients have a very good prognosis. Thus, so it is important to distinguish solid-pseudopapillary tumors from other tumors of the pancreas. PMID- 11885005 TI - Magnetic resonance hydrometry: non-invasive quantification of the exocrine pancreatic function. AB - AIMS: To show the ability of magnetic resonance hydrometry (MRH) to quantify the pancreatic secretion after secretin stimulation in order to distinguish between physiological excretion and reduced output in chronic pancreatitis. METHODS: MRH images were acquired in a 1.0-T-clinical scanner using a body-array coil and a heavily T2-weighted standard single-shot TSE sequence. Thirty-one patients (14 male/17 female) who routinely underwent ERCP for suspected choledocholithiasis (n = 22), recurring abdominal pain (n = 1), icterus (n = 6 and suspected pancreatitis (n = 2) were included. During the investigation 1 CU/kg BW secretin were administered intravenously. Secreted volume of fluid, start of secretion, achievement of a plateau of secretion and a combined score of these parameters (MRH score) were assessed and evaluated. Sensitivity and specificity were calculated for these parameters. RESULTS: 27 patients had no pancreatic pathology, and four suffered from chronic pancreatitis. Patients without pancreatic disorders produced a mean pancreatic fluid volume of 183 plus minus 86 mL, whereas patients with chronic pancreatitis secreted 61 +/- 39 mL. Secretion started after a mean time of 95 +/- 94 seconds (no pancreatic impairment) and 62 +/- 13 seconds (chronic pancreatitis). The MRH score achieved a high accuracy in the detection of chronic pancreatitis. CONCLUSIONS: Our study demonstrated the feasibility of measuring pancreatic output by MRH after stimulation with secretin. Moreover, a distinction between normal secretion and patients with chronic pancreatitis is possible. PMID- 11885006 TI - [Comparing the visualization of microcalcifications with direct magnification in digital full-field mammography vs. film-screen mammography]. AB - PURPOSE: To evaluate the conspicuity of microcalcifications in magnified mammographic views of preparations obtained with full field digital mammography (FFDM), film-screen mammography (FSM), and the DIMA technique. MATERIAL AND METHODS: Twelve preparations were examined by FFDM and FSM using 1.8 x magnification and DIMA using 7 x magnification. Parameter settings were identical for all three techniques. The number of visible microcalcifications was then determined for each modality by three radiologists. As far as possible, all preparations were X-rayed at 22 kV and 10 mAS. RESULTS: Altogether 9705 calcifications were counted (DIMA: 1609/1542/1534; FFDM: 1020/753/881; FSM: 901/643/822). The total number of microcalcifications identified with the DIMA technique was 4685 as compared to 2654 with FFDM and 2366 with FSM. The calcifications counted with FFDM and FSM thus corresponded to 56.6 % and 50.5 %, respectively, of those identified with DIMA. The differences between the groups were statistically significant (F-Test, p < 0.05). CONCLUSION: Significantly more calcifications are identified when magnified mammographic views of preparations containing microcalcifications are obtained with the DIMA technique compared to FFDM or FSM. FFDM depicts markedly more calcifications than FSM. This means one should increase spatial resolution. Digital mammography offers the potential for improved visualization of microcalcifications with advanced applications. PMID- 11885007 TI - [A differentiated approach to the diagnosis of pulmonary embolism and deep venous thrombosis using multi-slice CT]. AB - PURPOSE: To establish a differentiated protocol for multi-slice CT (MSCT) examinations in cases of clinically suspected pulmonary embolism (PE) using pulmonary CT-angiography (CTA) and indirect CT-phlebography (CTP). MATERIALS AND METHODS: 161 patients with suspected PE were examined using an MSCT (SOMATOM Volume Zoom; Siemens, Forchheim, Germany). After intravenous administration of 120 ml of contrast material, a thin collimation chest-CT scan was performed (120 kV, 100 mAs, collimation: 4 x 1 mm). If PE was present, or previous examinations and clinical signs suggested deep venous thrombosis (DVT), a CTP was subsequently completed. CTPs were performed using a 4 x 5 mm protocol (120 kV, 170 mAs). Venous phase scanning, starting from the pelvic crest, was completed in the popliteal fossa three minutes after contrast material injection. In 73 extremities, CTP were compared to the results of ultrasound, phlebography and autopsy. Scan ranges were documented in all patients. Cumulative doses were calculated for male and female subgroups. RESULTS: 62 patients in our series suffered from PE and in 47 of these patients deep venous thrombosis was seen additionally. Of the 99 patients without PE, 47 also received indirect CTP. CTP confirmed the suspicion and extent of DVT in 8 patients. Only in 2 of 39 patients (5.1 %) was previously unknown DVT found, despite the exclusion of PE. Regarding DVT, sensitivity was 94.3 % and specificity was 92.1 % for indirect CTP. Cumulative chest CT doses averaged 3.3 mSv for males and 4.2 mSv for females, the calculated CTP dosage was 9.3 mSv (according to ICRP 60). CONCLUSIONS: The examination protocol presented is suitable for clinical usage in patients with suspected PE. If PE is confirmed, indirect CTP is justified, so that detailed information of the venous system can be obtained. However, the relatively high radiation dosage of an additional CTP requires a strict indication regiment in patients with a negative CTA. PMID- 11885008 TI - [Clinical value of MRI concerning dissection of the supraaortic vessels]. PMID- 11885009 TI - [Evaluation of haemophilic arthropathy--a comparison of MRI and Pettersson score]. AB - PURPOSE: In order to compare score values, joint alterations in haemophilic patients on early and late prophylaxis were assessed and evaluated by X-ray radiography and magnetic resonance imaging (MRI) in a prospective study. PATIENTS AND METHODS: 24 joints (10 knees and 14 ankles) of 15 haemophilic patients, with at least one manifestation of bleeding (proven by sonographic assessment) were investigated. Radiological and MRI investigations were done after complete resorption of bleeding. For radiological evaluation a Petterson score (max. 13 points), and for MRI evaluation a modified MRI score (max. 13 points) according to Nuss et al. were used. RESULTS: Good correlation could be demonstrated between the number of joint bleedings and the degree of arthropathy. 16 joints with maximal 2 bleedings had no alteration on both MRI and radiological assessment. Joints with 3 bleedings had Pettersson score from 0 - 3 points and MRI scores of 2 points. Joints with greater-than-or-equal 4 bleedings had a radiological score between 7 and 12 points, MRI scores ranged from 3 to 8 points. CONCLUSION: Increasing severity of heamophilic arthropathy could be demonstrated initially by the MRI score and later by the Pettersson score depending on the number of bleedings. The MRI score describes initial joints alterations more precisely and earlier than the Pettersson score, allowing a discerning estimation of the degree and a follow-up of haemophilic arthropathy. PMID- 11885011 TI - [Percutaneous vertebroplasty]. AB - PURPOSE: To describe the procedure of percutaneous vertebroplasty and to present our first clinical results of patients treated for benign or malignant painful vertebral body disease. MATERIAL AND METHODS: We performed percutaneous vertebroplasty in 31 painful lesions of the spine. Liquid bone cement was injected into the affected vertebral body using fluoroscopic guidance through a bilateral transpedicular approach. Etiology of the bone disease was assessed by biopsy. Pain intensity was assessed before and 1 week after the procedure by standardized catalogue. RESULTS: Percutaneous vertebroplasty was performed in 17 thoracic and in 14 lumbar spine bodies of benign (n = 23) or malignant (n = 8) disease; no clinically relevant complications occurred. All patients reported significant pain relief 1 week after the intervention. One week after treatment, patients were pain-free in 15/31 vertebral bodies, and reported mild residual pain not necessitating narcotic medication in 16/31 cases. CONCLUSION: In accordance with the literature, percutaneous vertebroplasty proved to be a highly effective, minimal invasive interventional procedure to treat severely painful bone lesions of benign and malignant origin. PMID- 11885010 TI - [Endovascular gamma-irradiation for prevention or restenosis after angioplasty of femoropopliteal de-novo-stenoses-long-term results of a feasibility study]. AB - OBJECTIVES: To evaluate the performance and efficacy of endovascular irradiation after percutaneous transluminal angioplasty (PTA) of de-novo femoropopliteal stenoses in a pilot study. METHODS: 6 patients received non-centered endovascular irradiation (12 Gray at surface of the vessel wall) immediately after angioplasty of de-novo femoropopliteal stenosis, 1 patient was given centered endovascular irradiation using 192-iridium (12 Gray at surface of the vessel wall) Centered irradiation was considered for two other patients. Duplex sonographies and interviews were performed the day before and after PTA and after 1, 3, 6, 9, 12, 18, 24 months up to 4 years. Intraarterial angiography was performed in symptomatic patients. RESULTS: Non-centered endovascular irradiation was possible in all patients without problems or complications. Centered irradiation was not possible in two patients with the cross-over approach. One thromboembolic complication occurred during centered irradiation. Both restenosis and new stenosis at the edge of irradiated distance occurred in 1/7 patients. No other side effects were observed during follow-up. CONCLUSIONS: In our pilot study endovascular irradiation after angioplasty of de-novo femoropopliteal stenosis was possible with low rates of complications and restenosis and taking vessel anatomy into account. PMID- 11885012 TI - [Early results with a monorail-stent-balloon device for endovascular treatment of renal artery stenosis]. AB - OBJECTIVE: To evaluate the technical feasibility of a new monorail-stent-balloon device for treatment of renal artery stenosis (RAS). PATIENTS AND METHODS: During a study period of 18 months, 38 patients with proven RAS in 41 cases (hypertension n = 36, renal insufficiency n = 13) and indication for stenting (calicified ostial lesions n = 35, insufficient PTA n = 4, dissection n = 2) were enrolled into this prospective evaluation. Pre-mounted stents (Rx-Herculink(TM) 5 mm = 13, 6 mm = 34, 7 mm = 1) were implanted a transfemoral (n = 35) or transbrachial approach (n = 6). Mean grade and lengths of stenosis measured were 88 % plus minus 10 and 9 mm plus minus 5. RESULTS: Renal stent implantation was technically successful in all cases (100 %). In 7 cases a second stent had to be implanted to cover the entire lesion. The transstenotic pressure drop decreased from 88 mmHg plus minus 10 before to 1 mmHg plus minus 1.8 after the procedure. Remaining stenosis measured 0.7 % plus minus 4.2. Serum creatine levels decreased from 1.9 mm/dl to 1.5 mg/dl (n. s.), blood pressure decreased from 178/94 mmHg to 148/79 mmHg (p < 0.0001) after the intervention. Primary and secondary patency rates at 6 months were 72 % (Standard Error 9.8 %) and 77 (% (Standard Error 9.2 %), respectively. CONCLUSION: With the used monorail-stend-balloon device a technically easy, secure and exact renal stent placement is guaranteed, patency rates are similar to those described in the current literature. PMID- 11885013 TI - [Interactive direct volume rendering of CT-data: technical principle and applications]. AB - The advantages of interactive Volume Rendering Technique (VRT) for routine diagnostics of CT and particularly Multislice CT data are unquestionable. However, with apart from Virtual Endoscopy, where besides Shaded Surface Display, perspective Volume Rendering is used in some cases, VRT at present is only playing a minor role in routine diagnostics. One reason is time consuming calculation, so far requiring powerful computers. Due to the variety of possible parameter settings and the poorly predictable outcome of the resulting images, there also exists a certain discomfort in using the technique. In this paper the underlying principles of VRT are illustrated. Different parameter settings and their impact on the final images are explained. Possible pitfalls in VRT and also suitable parameter settings and indications for VRT are shown. PMID- 11885014 TI - [Evaluation of a new, portable ultrasound system in routine clinical use]. AB - AIM: Evaluation of a new, hand-carried ultrasound system (SonoSite 180) for routine use at a university hospital. METHODS: 101 routine ultrasound examinations were performed as bedside examinations on the wards or in the ultrasound centre by three experienced sonographers. The quality and results of examinations using the SonoSite 180 were evaluated in a standardized questionnaire. RESULTS: in 89 % of the ultrasound examinations, results were at least satisfactory. Even though the examiners felt some uncertainty due to the small screen, especially while performing abdominal scans, small lesions which were located far from the scanner were readily identified. CONCLUSION: Routine ultrasound examinations yield satisfactory results when performed with the new, hand-carried ultrasound system. The low size and weight of this ultrasound system and its good image quality makes it a very usefull tool, especially for examinations of patients at intensive care units and on hospital wards. PMID- 11885015 TI - [First clinical experience with a full-size, flat-panel detector for imaging the peripheral skeleton - Part II: Post-processing with a newly developed adaptive autowindow algorithm]. AB - PURPOSE: of the second part of the investigation was the evaluation of a newly developed adaptive autowindow algorithm in comparison to the system processing radiographs of the wrist and ankle to further optimize the image quality with softcopy reading. MATERIAL AND METHODS: All 120 radiographs of the wrist and all 100 radiographs of the ankle used in the 1st part of this paper were processed with the adaptive autowindow algorithm. The evaluation was again performed by 5 radiologists with softcopy reading. For the data analysis a variation of the Visual Grading Analysis (VGA) was used. RESULTS: Up to 19 % of the wrist radiographs and 2 % of the ankle radiographs processed with the system software had to be processed manually afterwards to get acceptable results. By the application of the adaptive autowindow algorithm a manual post-processing was no longer necessary. Highly significant (p less-than-or-equal 0.001) differences for all criteria to be evaluated were found for the wrist radiographs and in the case of the ankle radiographs for the bone contrast, the contrast in soft-tissue regions, the fine details in the bone and the artifacts, the adaptive autowindow algorithm performed always better than the system software. CONCLUSION: Using half of the exposition dose on a flat-panel detector, an optimized post processing leads to comparable or better results compared to the conventional film-screen-system concerning the image quality. PMID- 11885016 TI - [Determination of the distribution volume of contrast media solutions injected intrahepatically: pre-pilot studies for intratumoral gene therapy]. AB - PURPOSE: Determination of the intrahepatic distribution volume of two contrast media (CM) by CT-guided application in an ex-vivo and an in-vivo model (pig liver). MATERIAL AND METHODS: In pig livers ex-vivo and in-vivo, 131 CT-guided injections of two different CM (Imagopaque(R), Visipaque(R)) were performed using catheters and cannula with and without side-holes and documented by spiral CT. The distribution pattern was assessed visually: interstitial, subcapsular, vascular/tubular, the distribution volume was quantified using a density mask (thresholds 70/400 HE). RESULTS: Purely interstitial applications were achieved more frequently in-vivo than ex-vivo (p = 0.001). There were no relevant differences between the two CM. Catheters without side-holes led to more interstitial CM depots than catheters with side-holes (p = 0.005). The mean distribution volume was larger with catheters with side-holes (ex-vivo 103 cm(3), in-vivo 19 cm(3)) than with catheters without side-holes (ex-vivo 67 cm(3), in vivo 13 cm(3)) (p = 0,01). At the same time, the mean density with catheters with side-holes (102 HE) was lower than with catheters without side-holes (115 HE) (p = 0.005). CONCLUSION: Marked differences of the CM distribution volumes were observed between ex-vivo and in-vivo studies in the pig model. Catheters with side-holes are far superior to catheters without. PMID- 11885017 TI - [Adult, afebrile patient with rapidly growing tumor of the upper arm: Bartonella henselae as the etiology]. PMID- 11885018 TI - [Iatrogenic, symptomatic mesenterico-portal fistula]. PMID- 11885019 TI - Disconnection and migration of a subcutaneously implanted catheter system in a one-year-old boy. PMID- 11885021 TI - Treatment of malignancy by activation of the plasminogen system. AB - The blood coagulation mechanism in general and the plasminogen system in particular contribute to malignant growth and dissemination in complex ways. This article reviews the extensive literature that has accumulated over the past half century on effects of plasminogen activation on the natural history of experimental animal and human malignancy. Although the potent enzymes generated upon plasminogen activation may have a direct effect on tumor cells, it is more likely that their mechanism of action is related to disruption of the tumor cell extracellular matrix interaction. These observations suggest novel approaches to the experimental therapy of cancer. PMID- 11885022 TI - The coagulation system as a target for the treatment of human gliomas. AB - Coagulation activation in human gliomas may have two consequences: (1) activation of systemic coagulation reactions leading to the development of venous thromboembolic disease, and (2) stimulation of tumor growth and invasion. Anticoagulation in patients with gliomas, therefore, may not only prevent thrombosis but also have anticancer activity. Tissue factor and thrombin are appropriate targets for intervention, and several drugs are suitable for testing. Low-molecular-weight heparin and direct thrombin inhibitors are useful for reducing thrombin production and activity, and recombinant tissue factor pathway inhibitor and statins are examples of drugs that target tissue factor directly. This article reviews the implications of coagulation activation in human gliomas and provides a rationale for clinical testing of anticoagulants as part of a treatment strategy for this devastating human cancer. PMID- 11885024 TI - Platelets and cancer: implications for antiangiogenic therapy. AB - Thromboembolism is one of the most common causes of death in cancer patients. Among the most frequent thrombotic complications in patients with cancer are disseminated intravascular coagulation, thrombotic thrombocytopenic purpura, and thrombocytosis. Clearly, these complications arise as tumor cells interact with almost all components of the hemostatic system including platelets. Platelets participate in tumor progression by contributing to the metastatic cascade, protecting tumor cells from immune surveillance, regulating tumor cell invasion, and angiogenesis. Platelets contain one of the largest stores of angiogenic and mitogenic factors and the tumor vasculature is leaky, which allows platelets to come in contact with the tumor and deposit multiple angiogenic factors including vascular endothelial growth factor (VEGF) and thrombin to tumor cells, which in turn contributes to tumor progression. This article reviews the recent literature on how platelets contribute to tumor growth, angiogenesis, and metastasis. PMID- 11885023 TI - Effect of antihemostatic agents on experimental tumor dissemination. AB - There is now considerable evidence that the blood coagulation system plays an important role in the biology of malignant tumors. This evidence has been derived from a combination of clinical, biochemical, histological, and pharmacological observations that point to the possibility of favorably affecting the course of malignant disease with agents that interfere with blood coagulation pathways. For a number of years our laboratory has used experimental models of blood-borne metastasis to study the events that follow the introduction of procoagulant bearing tumor cells into the circulating blood. This article summarizes our experience with these models, which suggests that intravascular coagulation is a necessary prelude to lung tumor formation and that interruption of coagulation pathways in various ways may be an effective antimetastatic strategy. We have shown that anticoagulation with commonly used agents such as unfractionated heparin and warfarin (Coumadin) prevent tumor formation by limiting the ability of tumor cells to be retained in the pulmonary microvasculature. Binding of fibrin-coated tumor cells to activated platelets is essential for this retention and, therefore, treatment with potent antiplatelet agents such as abciximab is also effective. The predominant tumor procoagulant is tissue factor (TF), and direct targeting of this protein with concanavalin A, monoclonal antibodies, and tissue factor pathway inhibitor (TFPI) has provided compelling evidence that TF is an important determinant of tumor seeding in these experimental models. Collectively, our data provide strong support for the concept that some form of anticoagulant therapy would be a useful adjunct to existing cancer treatments. PMID- 11885025 TI - Anticoagulants in thrombosis and cancer: the missing link. AB - Many cancer patients reportedly have a hypercoagulable state, with recurrent thrombosis due to the impact of cancer cells and chemotherapy on the coagulation cascade. Studies have demonstrated that unfractionated heparin (UFH) or low molecular-weight heparin (LMWH) interferes with various processes involved in tumor growth and metastasis. These processes might include fibrin formation, binding of heparin to angiogenic growth factors such as basic fibroblast growth factor and vascular endothelial growth factor, modulation of tissue factor, and other mechanisms. Clinical trials have indicated a clinically relevant effect of LMWH as compared with UFH on the survival of cancer patients with deep vein thrombosis. Similarly, the impact of warfarin on the survival of cancer patients with thromboembolic disorders was demonstrated. Recent studies from our laboratory defined the role of an LMWH (tinzaparin), warfarin, anti-factor VIIa, and recombinant tissue factor pathway inhibitor in the modulation of angiogenesis, tumor growth, and tumor metastasis. PMID- 11885026 TI - Heparin inhibition of selectin-mediated interactions during the hematogenous phase of carcinoma metastasis: rationale for clinical studies in humans. AB - Classic studies indicate that the formation of tumor cell-platelet complexes in the blood stream is important in facilitating the metastatic process. Metastasis in animal models can be inhibited by heparin, and retrospective analyses of heparin use in human cancer have shown promise. However, most follow-up human studies using vitamin K antagonists have failed, and conclusive proof for other previously proposed mechanisms of heparin action is lacking. Carcinoma progression and metastasis are associated with overexpression of sialylated fucosylated mucins. Structurally similar molecules happen to be natural ligands for vascular adhesion molecules called the selectins. Heparin also happens to be a good inhibitor of P-selectin, which is expressed on activated platelets or endothelial cells. We have found that heparin blocks P-selectin-mediated interactions of endogenous platelets with sialylated fucosylated mucins on circulating carcinoma cells and that this reduces tumor cell survival. The use of more specific and selective P-selectin inhibitors will some day help to dissect the relative importance of this mechanism of heparin action in cancer. Meanwhile, we suggest that the failure of vitamin K antagonists to improve cancer prognosis should be ignored and that heparin therapy should be immediately revisited under this new paradigm. Unlike the suggestions in most previous studies, we propose that heparin use should be reexplored specifically during the interval from initial visualization of a primary tumor until just after its definitive surgical removal. A suggested clinical trial is outlined. PMID- 11885027 TI - Dynamic regulation of tumor growth and metastasis by heparan sulfate glycosaminoglycans. AB - This article focuses on the emerging views and concepts concerning the role of cell surface and extracellular heparan sulfate-like glycosaminoglycans (HSGAGs) in tumor biology. HSGAGs, found ubiquitously both at the cell surface and in the extracellular matrix (ECM), play a critical role in regulating tumor initiation, progression, and metastasis. The diverse biological functions of HSGAGs include the regulation of coagulation, growth factor signaling, cell adhesion, proliferation, and mobility. HSGAGs, depending on their location (anchored at the cell surface or soluble as free GAGs), the signaling molecules they associate with, and their fine structures, can either promote or inhibit the tumorigenic process. PMID- 11885028 TI - Improved cancer mortality with low-molecular-weight heparin treatment: a review of the evidence. AB - Work with low-molecular-weight heparins (LMWHs) continues to provide suggestions for survival advantages among patients with cancer diagnoses. Momentum is building in support of this theory through reports, the vast majority of which are derived from secondary analyses of clinical trials on the treatment of thromboembolism. The data retrieved from such studies that compare unfractionated heparin (UFH) with LMWH indicate that LMWH is equally beneficial if not more beneficial to cancer patients in terms of survival. In retrospective analysis, this improved life expectancy is not considered a result of reduced complications from thromboembolism. Thus, theories of antitumor effects of LMWH have developed, supported by evidence that most of the survival benefits are during long-term comparisons. Reports describing the effects of heparin in the setting of cancer have existed for over a half-century, although specific mechanisms for the marginal results seen thus far have yet to surface. Proposals for the most likely targets of the effective heparins include enzyme interaction, cellular growth modifications, and antiangiogenesis. PMID- 11885029 TI - Upper extremity deep venous thrombosis. AB - Upper extremity deep venous thrombosis (UEDVT) makes up approximately 1-4% of all episodes of deep venous thrombosis (DVT). Risk factors for UEDVT include central venous catheterization, strenuous upper extremity exercise or anatomic abnormalities causing venous compression, inherited thrombophilia, and acquired hypercoagulable states including pregnancy, oral contraceptive use, and cancer. Unexplained or recurrent UEDVT should prompt a search for inherited hypercoagulable states or underlying malignancy. Clinical presentations include arm, neck, and shoulder pain; edema; skin discoloration; tenderness; and venous distension. Because UEDVT is frequently asymptomatic until complications ensue, a high index of suspicion is required for patients with one or more risk factors for thrombosis. Pulmonary embolism and post-thrombotic syndrome are the most common sequelae of UEDVT. Early detection and treatment of UEDVT decrease complications, morbidity, and mortality. Compressive ultrasonography is an effective and economical means of confirming the clinical diagnosis in most patients. Traditional anticoagulant therapy of UEDVT is giving way to a multimodal approach involving transcatheter thrombolytic therapy followed by a minimum of 3 months of warfarin sodium anticoagulant therapy, venous decompression as needed, and balloon angioplasty with stenting for treatment of residual stricture. Low-dose anticoagulant therapy can safely and effectively mitigate the increased risk of UEDVT associated with the use of central venous catheters. PMID- 11885030 TI - A mutation hot spot for nonspecific X-linked mental retardation in the MECP2 gene causes the PPM-X syndrome. AB - We report here the genetic cause of the X-linked syndrome of psychosis, pyramidal signs, and macro-orchidism (PPM-X) in a three-generation family manifesting the disorder as a mutation in the methyl-CpG binding-protein 2 (MECP2) gene in Xq28. The A140V mutation was found in all affected males and all carrier females in the family. To date, descriptions have been published of two patients with independent familial mental retardation (MR) and two patients with sporadic MR who harbor this specific mutation in the MECP2 gene. This strongly suggests that A140V is a hot spot of mutation resulting in moderate to severe MR in males. A simple and reliable PCR approach has been developed for detection of the hot spot A140V mutation to prescreen any other unexplained cases of MR before further extensive mutation analyses. PMID- 11885031 TI - Genetic disease in offspring of long-term survivors of childhood and adolescent cancer treated with potentially mutagenic therapies. PMID- 11885032 TI - Skopje declaration on public health, peace & human rights, December 2001. PMID- 11885033 TI - Public health and peace. AB - The modern concept of public health, the New Public Health, carries a great potential for healthy and therefore less aggressive societies. Its core disciplines are health promotion, environmental health, and health care management based on advanced epidemiological methodologies. The main principles of living together in healthy societies can be summarized as four ethical concepts of the New Public Health essential to violence reduction equity, participation, subsidiarity, and sustainability. The following issues are discussed as violence determinants: the process of urbanization; type of neighborhood and accommodation, and consequent stigmatization; level of education; employment status; socialization of the family; women's status; alcohol and drug consumption; availability of the firearms; religious, ethnic, and racial prejudices; and poverty. Development of the health systems has to contribute to peace, since aggression, violence, and warfare are among the greatest risks for health and the economic welfare. This contribution can be described as follows: 1) full and indiscriminate access to all necessary services, 2) monitoring of their quality, 3) providing special support to vulnerable groups, and 4) constant scientific and public accountability of the evaluation of the epidemiological outcome. Violence can also destroy solidarity and social cohesion of groups, such as family, team, neighborhood, or any other social organization. Durkheim coined the term anomie for a state in which social disruption of the community results in health risks for individuals. Health professionals can make a threefold contribution to peace by 1) analyzing the causal interrelationships of violence phenomena, 2) curbing the determinants of violence according to the professional standards, and 3) training professionals for this increasingly important task. Because tolerance is an essential part of an amended definition of health, monitoring of the early signs of public intolerance is important. The vital interplay between the informed public and efficient administration, however, can only exist in an open society. The link between democracy and health of the people, and between public health and economic welfare is real. The Public Health Collaboration in South Eastern Europe (PH-SEE) evolved just in time to reconnect and strengthen disrupted professional networks in the region as a prerequisite of effective public health action. PMID- 11885034 TI - Health and peace. AB - Health and peace are closely linked. One cannot have one without the other. Although health and peace are desirable conditions, we human beings often thwart our best intentions to achieve and maintain them. War has profound impacts on human health. In addition to direct consequences, including the fact that 90% of all deaths related to recent wars were among civilians, war has several indirect consequences, including long-term physical and psychological adverse health effects, damage to the social fabric and infrastructure of society, displacement of people, damage to the environment, drainage of human, financial, and other resources away from public health and other socially productive activities, and fostering of a culture of violence. Many public health issues can be both a consequence and a cause of war, including infectious diseases, mental health disorders, vulnerability of population groups, disparities in health status within and among countries, and weakening of human rights. We, health professionals, can promote peace in many ways and facilitate this work by demonstrating our values, vision, and leadership. PMID- 11885035 TI - Contributing to Balkan public health: a school for Skopje. AB - The absence of social well-being and growing vulnerability are alarming for a large portion of people living in the Balkan countries. The Stability Pact is currently targeting the issue of social cohesion, which holds out promise for as yet unrealized development. Both the World Health Organization and the Council of Europe have called attention to the population vulnerability and growing disparity in health status between that region and Europe. Reversal of present trends demands the support of the international community and the strengthening of all public health institutions, human resource training, and population health research. Given the severity of the problem space of population vulnerability, these actions are more than ever indispensable to the health sector of the region. The paper describes an encouraging dialogue for Balkan health conducted by the National School of Public Health in Athens, Greece over the past decade and emphasizes the work of the newly created Public Health in South Eastern Europe (PH-SEE) Network (www.snz.hr/ph-see), which provides new opportunities for engagement in regional public health through Public Health Schools and Institutes. There is a need for public health curricula development and a closer linkage of all Schools with the Association of Schools of Public Health in the European Region. A curriculum for peace and public health is already under development in institutions in Athens, Greece; Mostar, Bosnia and Herzegovina; and Zagreb, Croatia. Soon to be added to the group of regional institutions is the School of Public Health in Skopje. It is a policy response to considerable need in a country showing pre-conflict conditions in the heart of the South Eastern Europe. Within the general framework of public health development, a School of Public Health in Skopje can be of great national benefit. Suggestions are made for its function under an umbrella of interdisciplinarity and autonomy, and the need to steer a path clear of medical dominance. According to a related mission statement, the School is to be implemented as an academic center of excellence and innovation, with the worthy purpose of improving the health of the population, with particular attention to the disadvantaged, underserved, and vulnerable. It can aid policy enactment, capacity building, and vulnerability research, promote the development of new training curricula for human rights and public health, and contribute to regional public health. The implementation of the School has a symbolism attached to it as a Balkan response for the elimination of the causes for political violence. PMID- 11885036 TI - Psychotrauma and reconciliation. AB - Our goal was to analyze the phenomenon of intergenerational transmission of trauma-related feelings and propose a way to alleviate this process through reconciliation of conflicted groups. The genesis of psychotrauma, with respect to the organism's defensive barrier being pierced by too strong external impulses, is discussed in terms of activation of the death instinct or by reactivation of childhood trauma. Genesis of the group trauma is explained in terms of chosen trauma, ie, activation of ancient national or other large group traumas, leading to the creation of malignant prejudices and hyper-activation of the social unconscious. Development of hatred, guilt, shame, and a need for revenge is illustrated with a number of examples from the 1991-1995 war in Croatia and the Holocaust, together with the influence they bear on an individual and a group. Therapy of psychotrauma could be a possible means of prevention of intergenerational transmission of traumatic emotions. Special attention is given to the retraumatization of Croatian war veterans in terms of the influence it exerts on future generations. A therapeutic model developed on the basis of eight years of clinical experience, designed specifically for veterans with posttraumatic stress disorder, is presented. The mechanism of intergenerational transmission of traumatic emotions as understood mainly through clinical experience in work with second-generation Holocaust victims is discussed. PMID- 11885037 TI - Neighbors and enemies: lessons to be learned from the Palestinian-Israeli conflict regarding cooperation in public health. AB - We aim to demonstrate on the example of recent Palestinian-Israeli collaborative projects how sustainable cooperation can be achieved despite unfavorable political atmosphere and continuing violent disputes. Palestinian and Israeli participants of the collaborative projects declared that their attitude and belief in coexistence was positively changed as a result of their experience in cooperative work. Their main motivations were a desire to contribute to the resolution of the conflict and to improve professional skills. The experience in Israeli-Palestinian conflict showed that the collaborative projects of higher chance of success and sustainability should address issues of high priority to the concerned parties, lead to demonstrable benefits in the immediate future, and preferably be organized by non-governmental organizations. Also, to secure long term success of collaborative projects, parity and symmetry should be maintained, as well as equal division of work and responsibility between the partners. Collaboration in public health field is first to be (re)established in conflict areas to alleviate suffering, minimize future health risks, and prevent further health deterioration. Thereby, public health can serve as a bridge to peace in areas of unrest or war. PMID- 11885038 TI - Developing partnership promotes peace: group psychotherapy experiences. AB - Partnerships are often optimal processes for interpersonal growth. The ability to have and keep a partner in mind should, therefore, be thought about and learnt. Although reciprocity, some symmetry, and mutual give and take are important aspects of partnerships, this article emphasizes a partners ability to process difficulties for the other as an aid to growth. The containment and elaboration of distress in partnerships is discussed using three examples of such potential relationships. The emotional beginning of a partnership, whether starting from love, working relationship or from hate, is the focus of the article. Individual, dyadic, and group aspects as separation-individuation and containment processes are described as contributing to partnership-building. The ability of a therapy group to process splitting and projecting phenomena are discussed. Co-therapists seem to have to work through painful conflicts between themselves to develop the therapists' containment abilities inside a functional partnership. Supervision may help process these emotional hardships. Within families, mothers could contribute to a better processing of their sons violence shared through infant dreams, which represent an effort to cope with inner and outer aggression. Growth promoting aspects of dream telling as potential partnerships in families and groups are discussed. Finally, partnership building between hating foes is exemplified by the efforts made by participants in Israeli and Palestinian peace dialogues. In groups, interpersonal development may be furthered by helping participants mutually contain and be contained, enabling partnership opportunities to grow after love and sympathy are over. PMID- 11885039 TI - Primary health care in complex humanitarian emergencies: Rwanda and Kosovo experiences and their implications for public health training. AB - In a complex humanitarian emergency, a catastrophic breakdown of political, economic, and social systems, often accompanied by violence, contributes to a long-lasting dependency of the affected communities on external service. Relief systems, such as the Emergency Response Units of the International Federation of Red Cross and Red Crescent Societies, have served as a sound foundation for fieldwork in humanitarian emergencies. The experience in emergencies gained in Rwanda in 1994 and Kosovo in 1999 clearly points to the need for individual adjustments of therapeutic standards to preexisting morbidity and health care levels within the affected population. In complex emergencies, public health activities have been shown to promote peace, prevent violence, and reconcile enemies. A truly democratic and multi-professional approach in all public health training for domestic or foreign service serves as good pattern for fieldwork. Beyond the technical and scientific skills required in the profession, political, ethical, and communicative competencies are critical in humanitarian assistance. Because of the manifold imperatives of further public health education for emergency assistance, a humanitarian assistance competence training center should be established. Competence training centers focus on the core competencies required to meet future needs, are client-oriented, connect regional and international networks, rely on their own system of quality control, and maintain a cooperative management of knowledge. Public health focusing on complex humanitarian emergencies will have to act in prevention not only of diseases and impairments but also of political tension and hatred. PMID- 11885040 TI - Challenge of goodness III: public health facing war. AB - Using moral and empirical analysis, we analyzed and discussed the role of public health in prevention of war as well as its function during and after the war. The idea is to develop a theory and new strategy in the spirit of public health to improve practices in preserving and strengthening peace, to be prepared for the future. The experiences from the last four wars in South Eastern Europe were ethical challenges to public health. We identified and described four models of public health practice in the past wars and conflicts. Based on the recent wars, the two new models, Professional Model and Peace Model, were developed and suggested as a new public health strategy in prevention and alleviation of the health burden of war. PMID- 11885041 TI - Human rights approach to health. AB - Adopting human rights approach to health carries many benefits, because it emphasizes the equality of all persons and their inherent right to health as the foundation of the health care system. It also argues that promotion and protection of health are fundamentally important social goals, focuses particularly on the needs of the most disadvantaged and vulnerable communities, balances individual needs with the common good, and so forth. However, it also raises some practical issues, such as organization of interdisciplinary education and work, and different use of the language, which often goes unacknowledged. The relationship between human rights and health is a reciprocal one, and can be beneficial or harmful. For the relationship to be beneficial and successful, the differences between human rights and public health approach to health, centered around the perspective taking, attitudes, and abilities of health professionals, need to be acknowledged and reconciled, and the need for interdisciplinarity adequately fulfilled. PMID- 11885042 TI - Minimum health indicator set for South Eastern Europe. AB - AIM: The Stability Pact includes a program for the development and reconstruction of training and research in public health for the countries of South Eastern Europe (PH-SEE). One of the identified priorities of national public health development is the definition of a Minimum Indicator Set for all countries of SEE. METHODS: A Task Force of the PH-SEE Network (www.snz.hr/ph-see) has proposed a Minimum Indicator Set on the basis of the list of the 224 indicators of the World Health Organization (WHO) Health for All (HFA) 21 strategy. The indicators selected follow the selection criteria as defined by expert groups of WHO and the European Commission. A meta-database describing the indicators should be established soon. RESULTS: A list of 32 indicators was agreed at a workshop in Ohrid, Macedonia, in September 2001. All indicators are included in the WHO HFA 21 indicator set. Some indicators are related specifically to the SEE post-war situation, such as indicators on suicide and homicide, literacy rate, average number of calories per person a day, and average number of persons per room. CONCLUSION: After principal agreement of the expert group on the list of indicators, further practical steps are necessary, especially testing the indicators and building a logistic network for realizing the Minimum Indicator Set. This includes a pilot phase, a revision of the Minimum Indicator Set after testing, responsibilities and timelines for data collection and data analysis, and transfer of the project into a continuous surveillance and monitoring system. PMID- 11885043 TI - Health monitoring of the migrant population in Northrhine-Westphalia, Germany: experiences, implications, and perspectives. AB - AIM: To describe the benefits and restrictions that emerged in health monitoring of the Northrhine-Westphalian migrant population. METHODS: Analysis of official register data, description and classification of benefits and restrictions, and systematic derivation of implications of general validity. RESULTS: The comparison of the native German and migrant populations revealed which health relevant fields with specific problems require political intervention and further research. The results clearly reflected strongly differing socio-demographic structures. Moreover, insufficiencies in the design of official statistics were found, which led to the formulation of general principles of an integrated system for the health monitoring of migrant populations. CONCLUSIONS: To serve as adequate data sources relevant for health monitoring that takes into account different dimensions of migration, official registers should fulfill certain requirements. Different indicators of migration and socioeconomic situation should be recorded, and classifications, such as national background and age, should be standardized in different statistical sources. PMID- 11885044 TI - Current health care system policy for vulnerability reduction in the United States of America: a personal perspective. AB - AIM: To raise questions about how the United States of America, which spends 1.3 trillion dollars on health care, conducts cutting-edge biomedical research, has the most advanced medical technology, and trains a cadre of highly competent health professionals cares for the most vulnerable members of its population. METHODS: Relevant statistical data were extrapolated from the most current statistical sources and research reports, and assessed in terms of existing practices and policies. RESULTS: The data clearly demonstrated that particular population cohorts -- the elderly, the poor, new immigrants, the homeless, the HIV-positive, and substance abusers -- were especially vulnerable to illness and its consequences. CONCLUSION: Since American medicine, despite all of its science, technology, and clinical competence, operates in a non-system, there is currently no efficacious approach to vulnerability reduction. To turn health care in the U.S. into a high quality, comprehensive, and cost-effective system, government officials, health care planners, and medical practitioners must address a series of fundamental social, economic, and political issues. What other countries, like those in South Eastern Europe, can learn from this is not to duplicate these mistakes. PMID- 11885045 TI - Refugee crisis in Macedonia during the Kosovo conflict in 1999. AB - The Kosovo refugee crisis in the Macedonia in 1999 was unique in terms of its unprecedented magnitude against its short duration (sharp increase and sudden decrease in refugee population), its high visibility in the world media, and attention received by donors. In the late March 1999, after the launch of the NATO air campaign against the Federal Republic of Yugoslavia, refugees from Kosovo began to enter Macedonia. Within 9 weeks, the country received 344,500 refugees. Aiming to provide an emergency humanitarian relief, United Nations, and international and national organizations together with the host country, donors, and other concerned parties coordinated and provided immediate assistance to meet the needs of refugees, including shelter in collective centers (camps) and accommodation in host families, nutrition, health care, and water/sanitation. The morbidity and mortality rates remained low due to the effective action undertaken by a great number of humanitarian organizations, backed up by strong governmental support. No significant epidemics developed in the camps, and there were no epidemic outbreaks during the crisis. Mortality rate of refugees was lower than in other emergency situations. PMID- 11885046 TI - Management of refugee crisis in Albania during the 1999 Kosovo conflict. AB - The report presents key data on Kosovo refugees in Albania during the 1999 crisis in Kosovo. In a three-month period, from March through May 1999, Albania received, accommodated, and cared for 479,223 officially registered refugees from Kosovo (FR Yugoslavia). Many foreign governmental and non-governmental organizations helped the Albanian government during the crisis. The Government cooperated with the organizations through Government Commission, which appointed a Special Coordinator to the Emergency Management Group that coordinated factors and actions in the field. A Health Desk was established by the Emergency Management Group to provide an overview of the health impact of the crisis upon refugees and domestic Albanian population. There were no serious outbreaks of infectious diseases, but the Health Desk registered 2,165 cases of diarrhea without and 14 cases of diarrhea with blood in the stool. Scabies and lice affected around 4% of the refugees. After the refugees returned to Kosovo, Emergency Management Group continued to coordinate the work on the rehabilitation of the refugee-affected areas. In this phase, humanitarian emergency work served as a bridge between emergency activities and normal development. PMID- 11885047 TI - International Organization for Migration: experience on the need for medical evacuation of refugees during the Kosovo crisis in 1999. AB - The International Organization for Migration (IOM) developed and implemented a three-month project entitled Priority Medical Screening of Kosovar Refugees in Macedonia, within the Humanitarian Evacuation Program (HEP) for Kosovar refugees from FR Yugoslavia, which was adopted in May 1999. The project was based on an agreement with the office of United Nations High Commission for Refugees (UNHCR) and comprised the entry of registration data of refugees with medical condition (Priority Medical Database), and classification (Priority Medical Screening) and medical evacuation of refugees (Priority Medical Evacuation) in Macedonia. To realize the Priority Medical Screening project plan, IOM developed and set up a Medical Database linked to IOM/UNHCR HEP database, recruited and trained a four member data entry team, worked out and set up a referral system for medical cases from the refugee camps, and established and staffed medical contact office for refugees in Skopje and Tetovo. Furthermore, it organized and staffed a mobile medical screening team, developed and implemented the system and criteria for the classification of referred medical cases, continuously registered and classified the incoming medical reports, contacted regularly the national delegates and referred to them the medically prioritized cases asking for acceptance and evacuation, and co-operated and continuously exchanged the information with UNHCR Medical Co-ordination and HEP team. Within the timeframe of the project, 1,032 medical cases were successfully evacuated for medical treatment to 25 host countries throughout the world. IOM found that those refugees suffering from health problems, who at the time of the termination of the program were still in Macedonia and had not been assisted by the project, were not likely to have been priority one cases, whose health problems could be solved only in a third country. The majority of these vulnerable people needed social rather than medical care and assistance a challenge that international aid agencies needed to address in Macedonia and will need to address elsewhere. PMID- 11885048 TI - Children's well-being after the war in Kosovo: survey in 2000. AB - AIM: To assess special health and psychosocial needs of Albanian children in Kosovo shortly after the dramatic ethnic conflict in this part of former Yugoslavia in 1999. METHODS: The survey included representative samples of school age children (n=813), parents (n=41), and teachers (n=31) from six public schools in Prishtina and surrounding area. The measuring instruments included a standardized inventory of children's coping behavior in stressful situations (Ryan-Wegner Coping Style Inventory, SCSI), and survey questionnaires for children, parents, and school teachers, which were also used in a parallel study in Bosnia and Herzegovina (Sarajevo) on comparable survey samples. The study was accomplished in April 2000, ie, only a few months after the crisis in Kosovo. RESULTS: At the time of the survey, many children in Prishtina and surrounding area lived in unhealthy and dangerous physical environment. There were frequent lack of electricity (74%), lack of safe drinking water (68%), garbage on the streets (63%), and firearms, explosive devices, and mine fields in close environment (45%). Many of them showed signs and symptoms of ill health, including frequent headaches (60%), stomach ache (41%), frequent high fever (32%), and sleeplessness (18%). Most of them felt unsafe on the streets (61%). Many of them had rather unhealthy eating habits, such as not having breakfast regularly (16%) or not having a morning snack (60%). Three major groups of stressors were identified as having impact on children's health and psychosocial well-being in Kosovo, as follows: 1) lack of cultural and social security resources at home and in the community at large (20% of common variance explained); 2) poor physical and mental health conditions (14% of common variance); and 3) school-related stressors (11% of common variance). Similarly, parents and teachers also lived and worked under stressful life conditions. Many parents feared the impact of traumatic war experiences on children's health (54%), and school teachers noticed high rates of children's learning and behavioral disorders (84%). Factor analysis of the SCSI proved the hypothesis that in stressful situations children tend to use two major coping strategies: either active, ie, object-focused coping (13% of variance explained) or passive, ie, self-focused coping (10% of variance explained), the later being more typical for younger children. The pattern of stressors and coping behaviors were similar to stressors impacting physical and mental health of children in Sarajevo, although there were a number of culture-specific differences. CONCLUSION: Environmental, educational, and social conditions must be respected in assessing impact of war and conflict on children. Promotion of solidarity, tolerance, and mutual support among children from different ethnic and cultural backgrounds should be encouraged. PMID- 11885049 TI - Medical aspects of the mass-scale civilian casualties at Sarajevo Markale market on August 28, 1995: triage, resuscitation, and treatment. AB - The siege of Sarajevo, which holds an important but tragic position in the history of war, lasted for more than 45 months, from April 6, 1992 to March 19, 1996. Shelling of the Markale city market on August 28, 1995, was the attack with the largest number of civilian casualties. There were 23 persons killed on the spot. Another 104 were injured, of whom 15 died immediately after the explosion or during surgery, and 4 died a week later. Transport to the hospital was provided mostly by other civilians and resuscitation on the spot was not attempted. The triage of the wounded was conducted at the Kosevo University Hospital Center and State Hospital by teams of surgeons and anesthesiologists. Out of 104 wounded, 94 were treated at several different Surgery Departments and the Emergency Department. There were 85 survivors among the wounded. This incident once again illustrates the importance of timely adequate triage and resuscitation after mass-scale injuring, which can increase the chances of survival. PMID- 11885050 TI - Psychological status of Sarajevo children after war: 1999-2000 survey. AB - AIM: To make a survey of children's health and psychosocial needs after the 1992 1995 war in Sarajevo, Bosnia and Herzegovina. METHODS: Representative samples of school-age children (n=310) from 6 public schools in the Sarajevo Canton, their parents (n=280), and teachers (n=156) were surveyed by means of self-administered questionnaires and standardized psychometric scale (Ryan-Wengers Schoolagers Coping Strategies Inventory). The survey was conducted in October-November 1999, approximately four years after the war. RESULTS: At the time of survey, well being of children in Sarajevo was still heavily impacted by many various unhealthy life conditions and psychosocial stressors. Many school-age children lived in unhealthy and dangerous environment, including overcrowded living conditions (40%), unsafe playgrounds (68%), and no access to sports fields (52%). Most felt unsafe on streets (74%), many (73%) coped with one or more school problems, and even 84% were ill at least once during the past 12 months. General poverty was the prime stressor (common variance explained: 23.5%), followed by school- and health-related risks (common variance explained: 17.0%). At the third place were family-associated risk factors impacting children's health and development, such as overcrowded living conditions and lack of social support within their own family (common variance explained: 10.5%). Parents and teachers also lived and worked in stressful life conditions and were concerned for both their children's and their own well-being. Despite all that, most children tended to use healthy strategies in coping with stressful events in their everyday lives. CONCLUSION: The reinforcement of children with positive (healthy) coping skills and strengthening of their social support networks seems to be the most important intervention strategy to help the war-traumatized children in Bosnia and Herzegovina. PMID- 11885051 TI - Psychiatric help to psychotraumatized persons during and after war in Croatia. AB - AIM: To present organization of psychosocial support and treatment of traumatized persons during and after the 1991-1995 war in Croatia. METHOD: Description of application and results of community-based National Program of Psychosocial Help to War Victims, and retrospective analysis of hospitalizations for psychotrauma at the National Center for Psychotrauma, Dubrava University Hospital. RESULTS: During the 1991-1995 war in Croatia, the population faced severe traumatic events, and the need for organized psychosocial help to traumatized persons was great. Government has established the network of psychosocial help in 1994. This was the beginning of the National Program of Psychosocial Help to War Victims. As a strategy in building a social support, the pyramidal model consisting of five levels of psychosocial help was used. The levels were the following: 1) preventive programs in mental health, 2) nonspecific psychological help in community, 3) basic psychological help, 4) psychiatric institutionalized help and specific psychological help, and 5) national coordination and operative planning. During the war, the work primarily centered around community-based approach, satisfying the current needs of the traumatized people, such as food, medication, and clothes, and providing crisis psychological interventions and urgent psychiatric help. In 1999, this organizational scheme was replaced by the establishment of the National Center for Psychotrauma and four Regional Centers for Psychotrauma in Zagreb, Rijeka, Osijek, and Split. In the post-war period, the emphasis was on psychological and social help in the community and on institutionalized approach to treatment of psychotrauma. According to the data of the Croatian Institute of Public Health, in the 1998 and 1999 there were 1,744 and 2,415 hospitalizations due to PTSD and related disorders, respectively. CONCLUSION: The community-based approach remained central in dealing with psychotraumatized persons during and after the war. In the post-war period, the emphasis was put on non-specific and specific psychological and social help in the community, with institutionalized approach to the treatment of PTSD and related disorders, whereas satisfying the current needs (food, medication, clothing), intervention in the crisis situations and urgent psychiatric interventions were of prime importance during the war. PMID- 11885052 TI - Health care relief to neighbors: Split University Hospital during the 1991-1995 war in Bosnia and Herzegovina. AB - AIM: To assess the workload of the Split University Hospital during the war and its role in providing help to the neighboring countries. METHODS: We reviewed all available records of patients admitted to the four (out of 15) departments: General Surgery, Traumatology, Dermatovenerology, and Pulmonology. The files of 37,821 patients (78% of total number) treated during 1990-1995 were analyzed. RESULTS: The workload of the hospital paralleled the political crisis in Bosnia and Herzegovina (BH) -- the number of patients from BH increased more than 10 fold between 1990 and 1993, including during the time of armed conflict between BH Croats and Bosniaks (1993-94). Among them, there were 84% of ethnic Croats and 16% of ethnic Bosniaks. The hospital spent US$6.2 million (18% of total costs) on the treatment of BH citizens. Approximately two thirds of BH citizens (62%) were treated at one of the surgical departments, and approximately one third of Bosniak patients were young males, admitted for treatment of war-related injuries. CONCLUSION: The Split University Hospital took a large burden of managing BH citizens, despite the armed conflict of Bosnian Croats and Bosniaks, indicating the high professionalism of the hospital staff and management. Such attitude can contribute to peace and post-war reconciliation in the region. PMID- 11885053 TI - Linking objects in the process of mourning for sons disappeared in war: Croatia 2001. AB - AIM: Mothers use linking object to externalize the complex aspects of their relationship to the loss of their child. We analyzed the linking objects that mothers kept in memory of their sons who disappeared in the 1991-1995 war in Croatia or whose remains were uncovered and identified long time after they had gone missing. METHOD: The case study of disturbed mourning included 26 mothers of Croatian soldiers from Croatian Osijek-Baranja County who went missing in war or whose remains were recovered and identified long after they had gone missing. The mothers were selected independently by the president of the Association of Families of Missing and Detained Croatian Soldiers and agreed to participate in the study in 2001. They were interviewed in their homes, their testimonies were recorded, and photographs of the linking objects taken. Linking objects were classified according to the Volkan's four-group classification. RESULTS: Out of four Volkan's groups of linking objects, we identified the objects belonging to the first three. Those were 1) objects that had been worn by the deceased (clothes, wrist-watch, ring, or glasses), (6/26); 2) objects that could be viewed in the psychoanalytic sense as an extension of the body of the disappeared or dead person, such as a camera (4/26); and 3) objects with realistic or symbolic resemblance to the deceased, usually a photograph (8/26). None of the examined objects belonged to the fourth Volkan's group (objects at hand when the news of the death came or objects present at the funeral, things that could be considered last-minute objects, ie, related to the moment when the deceased was last seen alive). However, 8/26 objects formed a new hitherto undescribed group. Mothers used such objects to create a memorial shrine to their sons. A photograph of the missing person or person whose remains were identified long after he had gone missing occupied a central place at the shrine, and was surrounded by other symbols of the Catholic iconography (Virgin Mary, crucifix), flowers, and candles. The memorial shrine to the beloved son who disappeared was always located in the room where the family spent most of their time and/or where guests were received (living room or kitchen). CONCLUSION: We found three out of four original (Volkan's) groups of linking objects, but also an additional one, hitherto undescribed, comprising objects used for designing a memorial shrine to the deceased. This could be viewed as an expression typical of Christian, mid European Croatian culture and tradition. PMID- 11885054 TI - Bulgarian population in transitional period. AB - In the transition period from a communist to market-oriented economy, Bulgaria faces several public health challenges. One of them is the decline in population (estimated fall from current 8.25 million to around 6 million in 2045), mainly due to emigration and pronounced fall in fertility. Infant mortality is still relatively high (over 15/1,000 live births), and the incidence of tuberculosis is on the rise. Total mortality shows a steady upward trend from 12.1/1,000 in 1990 to 14.3/1,000 in 1998. Trends in ischemic heart disease are comparable to those in other Central and Eastern European countries, but stroke mortality is notably higher. This calls for detailed epidemiological studies of risk factors, such as salt consumption, as well as preventive programs for detection and control of high blood pressure. The problems of smoking and alcohol abuse should be addressed by a coordinated public health and legal measures. PMID- 11885055 TI - Burden of tuberculosis in Afghanistan: update on a war-stricken country. AB - AIM: To review Afghans Tuberculosis (TB) Control Program and assesses the impact of disruption induced by the war in Afghanistan. METHODS: National TB control program of Afghanistan was reviewed in terms of its milestones, achievement parameters, and potential barriers. Information and data were collected by review visits to the Ministry of Health and health facility survey of non-governmental organizations working for TB control in Afghanistan. Local and international literature was consulted. RESULTS: Mortality and morbidity figures due to tuberculosis remained alarmingly high in the last two decades, especially among women. Current estimates show that the incidence of active TB cases is 278 per 100,000 and mortality mounts to 15,000 cases per year. The epidemiological profile reflecting the situation of Afghans inside and outside the country is extremely deplorable. The situation has worsened due to the cessation of TB control activities during the war. Compliance of patients and access to the treatment has become very difficult in an emergency situation. Similarly, an increasing number of TB cases among Afghans refugees in Pakistan have also been observed. Overcrowded refugee camps and lack of treatment facilities increases manyfold the risk of further transmission. CONCLUSION: TB is a major public health threat inside and outside war-stricken Afghanistan. TB control activities need prompt attention of health authorities in reestablishing TB control network. World Health Organization's guidelines and nationwide Directly Observed Treatment Short Course strategy should be adopted and sufficient resources allocated. It is vital to build a peaceful environment with a viable and durable alliance of local and international donors in the fight against TB. PMID- 11885056 TI - Church bells. PMID- 11885057 TI - Public health and quality of care improvement. PMID- 11885058 TI - Dispelling myths about depression. PMID- 11885059 TI - Human immunodeficiency virus (HIV) disease in persons with severe mental illnesses. AB - Extant research has found alarming rates of the human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) in persons with severe mental illnesses (SMI), with seroprevalence rates ranging from 4% to 23%. However, persons with SMI have received less attention than any other group that has been disproportionately affected by HIV/AIDS. As such, preventing and treating HIV in persons with SMI has been indicated as one of the highest priority research initiatives by the NIH Office on AIDS research. This article reviews current research findings and discusses the nursing research and practice implications for assessment, prevention, and treatment of HIV in this vulnerable population. PMID- 11885060 TI - Issues of women dually diagnosed with HIV infection and substance use problems in the Carolinas. AB - A growing number of women are being dually diagnosed with HIV infection and substance use problems. Forty-two percent of all women diagnosed with AIDS have been infected through injection drug use. Many more women with HIV are exposed to nonintravenous drugs that potentially affect their quality of life and illness experience. This study sought to identify from the perspective of women factors that most influenced their ability to obtain treatment for their HIV infection and control their substance use. A focus group approach was used for data collection. Twenty-five HIV-infected women participated in one of four focus groups. Women were asked to identify and discuss their concerns and needs related to HIV/AIDS and substance use. Twenty-four women were African-American; one was white. All the women reside in South Carolina or North Carolina. Each focus group session was audiotaped and transcribed. Content analysis, following Krippendorff's (1980) methodology, was used to analyze the data. Five themes emerged: 1) AIDS as a life-altering event; 2) spirituality; 3) mental health issues; 4) barriers to health care services; and 5) environmental influences. It was concluded that the coexistence of HIV and substance abuse adds to the complexity of women's treatment needs. For these women, an HIV diagnosis can serve to alter their lives either positively or negatively. Dually diagnosed women have unique needs that require integration of physical and psychosocial interventions. These women may benefit from the services of psychiatric or mental health nurse practitioners who have the skills necessary to address the many psychosocial issues women face as well as provide physical treatment. Additionally, drug treatment services need to be expanded and made more comprehensive. Drug treatment programs need to be developed specifically for women, and these services need to be made accessible to poor women with substance abuse problems. Further, drug treatment programs need to provide comprehensive services that can appropriately integrate the treatment of HIV disease and substance abuse. PMID- 11885061 TI - The mental health clinical nurse specialist and the "difficult" patient: evolving meaning. AB - Twelve mental health clinical nurse specialists (CNSs) working in outpatient mental health settings were interviewed and asked to describe situations where they had experienced difficult client behavior. Study data, analyzed via the grounded theory method, revealed the basic social process of Evolving Meaning. Evolving Meaning signifies change over time, based on both Enhancing Experience and Expanding Understanding. The phases of Personal Meaning, Negotiating Meaning, and Illuminating Meaning were recognized as central to the basic social process of Evolving Meaning. The study findings emphasized the importance of the nurse client relationship process. Although the CNS participants did describe client behavior that created difficulty for them, the CNS-client relationship was viewed as being interactive and subsequently, difficult behavior was viewed within the context of that relationship. Clinical supervision was recognized as an essential component of outpatient mental health CNS practice, even by the more experienced study participants. In this study, positive components of clinical supervision included validation, insight, and system support. A surprising finding of the study was the intrusive behavior of clients, including stalking of some CNSs, their family members, or both. PMID- 11885062 TI - Coping styles, stress levels, and the occurrence of spontaneous simple reminiscence in older adult nursing home residents. AB - This article reports the results of a preliminary descriptive correlational study developed to evaluate the relationships among coping styles, stress levels, and the occurrence of spontaneous simple reminiscence in a group of older adult nursing home residents. Several important issues that will affect future studies of these relationships were identified during the implementation and evaluation of this study. Identified stressors did not appear to contribute to the use of spontaneous simple reminiscence and subject recruitment was challenging. Suggestions for responding to these issues in future studies are offered. PMID- 11885063 TI - Moderating and mediating effects in causal models. AB - This article explains causal relationships in conceptual models of mental health phenomena. Direct, moderating, mediating, and reciprocal effects among variables are defined, appropriate statistical analyses are described, and the correct interpretations of moderating versus mediating effects are discussed. Examples are provided that will help the reader to distinguish between moderating and mediating effects. PMID- 11885064 TI - "So that our souls don't get damaged": the impact of racism on maternal thinking and practice related to the protection of daughters. AB - Racism influences the conceptualization of motherhood and the practice of mothering. Narrative analysis was completed on 246 stories collected through five focus groups. Twenty-five African American women participated in the study. The results indicate that living in a racist society profoundly impacts the maternal thinking and practice of African American women in relation to protecting their daughters. Protection of children was viewed as a communal responsibility. Issues related to the provision of safe and nurturing physical, aesthetic, and spiritual environments are addressed. The influence that maternal responsibilities have on women's own health is also discussed. Suggestions are provided for mental health providers who wish to work more effectively with African American women. PMID- 11885065 TI - Recent developments in psychosocial interventions for people with psychosis. AB - The advent of drug treatments for psychotic illness in the 1950s, along with changes in social policy, heralded the move from institutionalized care to community care. Over the last decade, there have been research developments in the use of psychological techniques to manage psychotic symptomatology, particularly in the realms of cognitive behavioral therapies. There is growing evidence to suggest that psychological treatments can offer an adjunct or even an alternative to traditional medical treatments for patients with schizophrenia and other serious mental illnesses. Opportunities have arisen for mental health nurses to learn these new approaches to caring for people with enduring mental illnesses. The impact of psychological interventions in the treatment of psychosis and the implications for mental health nursing practice are discussed. PMID- 11885066 TI - African American mothers' responses to hospitalization of an infant with serious health problems. AB - PURPOSE: To describe African American mothers' experiences related to the hospitalization of an infant with serious health problems. DESIGN: Retrospective descriptive. SAMPLE: 19 African American mothers with premature and term infants who were hospitalized at birth for serious health problems related to sequelae of prematurity or birth defect. MAIN OUTCOME VARIABLE: African American mothers' recollections about the hospitalization of their seriously ill infant. RESULTS: The mothers worried primarily about when the baby could go home. Their greatest source of stress was separation from the infant. Seeing their sick infant was also stressful and evoked shock, fear, denial, guilt, and helplessness. Mothers sought hope by seeking information and cues from the infant and by praying to God. Mothers established a relationship with their infant by visiting regularly and by learning how to care for him. Some mothers feared getting attached to an infant who might die. Mothers' highest source of satisfaction was support from the health care team. PMID- 11885067 TI - Neonatal ethical decision making: where does the NNP fit in? AB - Neonatal nurse practitioners are frequently confronted with ethical dilemmas in the NICU. This article reflects on the historical basis of ethical decision making and the issues some novice NNPs face today regarding their participation in the decision-making process. It examines various educational strategies that can be used to help NNPs to develop the skills they need to participate collaboratively in ethical decision making in the NICU. It concludes with recommendations for further research. PMID- 11885068 TI - More clinical research needed for children. PMID- 11885069 TI - Wide range of skills make research nurse role a challenge. PMID- 11885070 TI - So happy together. Co-bedding multiples boosts growth and development, enhances bonding. PMID- 11885071 TI - Update on reimbursement issues faced by neonatal nurse practitioners. PMID- 11885072 TI - Securing ET tubes. PMID- 11885073 TI - Drug-of-abuse testing in the neonate. AB - Drug-exposed mothers and infants continue to challenge maternal and infant health care resources. Complex family problems, polydrug abuse, long-standing drug use, and resistance to change may complicate intervention and treatment. Maternal motivation to change may be significant at the time of a child's birth, however. Identification of the infant exposed to drugs of abuse may provide an opportunity for health care providers to intervene at a time when the mother is open to assistance in addressing her drug abuse problem. Performing drug testing on the neonate without providing the necessary family assessment, referral, and follow up services limits the usefulness of the test. Awareness of community resources, referral mechanisms, and involvement of social services provides an integrated approach to the family and a better outcome for the infant. PMID- 11885074 TI - Newborn percutaneous absorption: hazards and therapeutic uses. PMID- 11885075 TI - Autosomal trisomies: what neonatal nurses need to know. AB - Autosomal trisomies are associated with major congenital malformations that may result in prolonged hospitalization of the newborn. Knowledge about these chromosomal abnormalities is important for nurses in neonatal practice. This article identifies the causes and manifestations of most of these trisomies: trisomy 13 (Patau syndrome), trisomy 18 (Edwards syndrome), and trisomy 21 (Down syndrome). More detailed description of the manifestations, associated abnormalities, and outcomes of the most common of these, trisomy 21, is provided. PMID- 11885076 TI - Neonatal air leaks: pneumothorax, pneumomediastinum, pulmonary interstitial emphysema, pneumopericardium. PMID- 11885077 TI - Screening of antiadherent activity on Streptococcus sobrinus culture. AB - The mutans group of streptococci is considered to play a key role in the etiology of dental caries. We have evaluated the ability of different substances to prevent dental plaque formation without affecting Streptococcus sobrinus viability. Viable organisms were detected as CFU/mL in agar plates and bacterial adherence was assessed by dry weight. We studied 23 compounds and we demonstrated that phenyl salicylate, phenylmercuric nitrate and potassium iodate are more effective to inhibit adhesion without showing antibacterial activity. PMID- 11885078 TI - Chronodynamic evaluation of the stages of osseointegration in zirconium laminar implants. AB - Osteogenesis occurs throughout all stages in life, due to both bone turnover and reparative processes. Thus, osseointegration (OI) can be described as the final step in a cascade of processes involved in bone healing in relation to implants. Ten groups of 5 Wistar rats each (mean = 90 g b.w.) were used. Under ether anesthesia a zirconium laminar implant was placed in the tibia following the method previously described by our laboratory (Cabrini et al Imp Dent 2:264-7, 1993). The animals were killed at Ohs, and 1, 2, 3, 6, 7, 10, 12, 14, and 60 days post-implantation. Tibiae were resected, radiographed and processed for their embedding in methyl methacrylate. Three sections, perpendicular to the major axis of the tibia, were obtained per implant and histologic and histomorphometric studies were carried out. Volume occupied by blood clot, woven bone, percentage of OI and OI bone tissue thickness, were determined. Histologic and histomorphometric studies as function of time revealed: a) at 6 days the presence of non-osseointegrated woven bone around the device is evident increasing in volume from 7 to 10 days post-implantation, and disappearing from day 12 to 14., b) at 14 days after implantation lamellar bone formation on the surface of the zirconium implants (OI) is noticeable. Additional bone growth is observed after 60 days. This study enables quantification of peri-implant reparative process response in an unloaded, necrotic trabeculae free model showing, in the different phases of the osseointegration process, the role of the blood clot and of the appearance and disappearance of woven bone and the final stages of osseointegration. Further investigation will allow comparison of results obtained under the effect of local and/or systemic factors that might affect osseointegration. PMID- 11885080 TI - A culture medium for simultaneous counts of Streptococcus mutans and lactobacilli in saliva. AB - The use of a single culture medium that allows the isolation and counts of both Streptococcus mutans and lactobacilli could be of great value in microbiological diagnosis, control and evaluation of prevention programs that are nowadays employed in Odontology. To date there is no method that allows the simultaneous counts of lactobacilli and S. mutans in oral samples using a single culture medium. A single culture medium would allow for a more exact diagnosis of cariogenic risk and activity and a reduction in costs and processing time. We here in propose the selective-differential LAPTg 7% sucrose medium to differentiate oral streptococci and lactobacilli according to colony morphology and dextran production. The choice of this medium was the result of testing culture media such as MRS Agar, Elliker Agar and modified LAPTg Agar. PMID- 11885079 TI - Effects of xylitol, sorbitol and fluoride mouthrinses on glucose clearance in adolescents. AB - The present work describes and analyzes the results of a randomized clinical trial on 98 healthy adolescents (age 18 +/- 0.7 years) in order to evaluate the effects of a 14 days treatment with mouthrinses containing xylitol (0.2%; 0.5% and 1%), sorbitol (1%), NaF (0.1% respectively) on salivary glucose clearance. In all volunteers oral glucose clearance followed an exponential curve as a function of time, which fitted almost exactly to the equation log Ct = log Co - bt from 1 to 16 minutes after sugar rinsing. Xylitol treatment provoked an increase in oral glucose clearance, which was proportional to its concentration in the mouthrinse formula. The average AUC (area under curve) decrease was 9.1% in subjects rinsing with 0.2% xylitol; 21.5% with 0.5% xylitol and 40.0% with 1% xylitol. 1% sorbitol or 0.1% NaF did not modify any of the pharmacokinetical parameters over the same treatment time. The mouthrinses containing 1% xylitol and 0.1% NaF produced the same results as 1% xylitol alone on oral glucose clearance. No significant changes in the salivary flow rate nor in oral health parameters were observed concomitant to the faster oral glucose clearance by xylitol treatment. Since the sugars salivary clearance is part of a process intended to prevent dental caries, our results suggest that xylitol adds another mechanism of action to its well known cariostatic and anticaries properties. PMID- 11885081 TI - Mitis salivarius-bacitracin 10% sacarose agar for oral streptococci and Streptococcus mutans counts. AB - The MSB Agar (mitis salivarius-bacitracin) 20% sacarose medium is frequently used for the isolation and count of total streptococci and Streptococcus mutans. Although it is considered a selective culture medium for this micro-organism, S. mutans recovery in this medium is much lower than in this Mitis Salivarius Agar (MSA). Because the number of S. mutans in saliva is used for estimating caries risk and activity from a microbiological stand point, the aim of this work was to find a modification of the MSB 20% sacarose medium so that it would offer not only selectivity in the isolation but also maximum recovery. This would detect people at risk more efficiency and would evaluate the preventive odontological treatments more accurately. The results show that: 1) the greatest recovery of total streptococci and S. mutans is obtained in the MSB 10% sacarose medium, 2) S. mutans must be incubated in aerobiosis and the total streptococci in a candle jar (10% CO2). MSB 10% sacarose medium is proposed as a choice medium for the microbiological estimation of cariogenic risk and activity, to detect infection levels and evaluate preventive odontological treatments. PMID- 11885082 TI - Iron overloading inhibits endochondral ossification. AB - The development of bone disease in patients with chronic renal failure is well known. Renal patients frequently suffer anemia and iron oral therapy and/or transfusions are used to treat them. Recent findings show that iron could be a factor that provokes bone lesions but the alterations it causes are not well known. The aim of this work was to study the effect of iron intoxication on endochondral ossification, a recognized model for bone growth evaluation. Male Wistar rats weighing 250-300 g were used. They received 88 mg of dextran-iron per day intraperitoneally during 34 days. The experimental and control groups were killed on day 34. A histomorphometric study of the endochondral plate was performed on histologic sections of tibiae. The results obtained show that iron overloading inhibits endochondral ossification. PMID- 11885083 TI - Scientific presentations and publications on odontological research in Argentina. AB - The results of odontological research which are presented at the annual meetings of the Argentine Division of the International Association for Dental Research (A.D.I.A.D.R.) are proof of the scientific production of this country in this area. An analysis of the presentations allows for the quantitative evaluation of the activity of the area. A deeper appraisal of the reality of research, involves the analysis of quality and publication efficiency. A useful indicator is the relationship between the quantity of the presentations and subsequent publications (Publ./Pres. Ratio) in Journals with peer review. In 1990, the authorities of the Division presented an evaluation of the 10 previous years (Acta Odont. Latinoamer. 7(2):39-46, 1993). The current Board of Directors has considered timely to update that information. With this aim in mind the presentations at A.D.I.A.D.R. over the period 1990-1995 were considered. Employing the authors index of the A.D.I.A.D.R. meeting we searched for possible publications in Medline. The references were compared with the data from the presentations, disregarding those which had not been communicated previously in the Division. The data obtained were grouped according to Research Center and subject area. The Publ./Pres. Ratio was calculated. The time to publication and language of publication were considered. Of a total of 506 presentations, 61 were published, Ratio Publ./Pres. 1:8 (12%). Considering each Center individually the ratio was 1:6 for the University of Buenos Aires (UBA), 1:13 for the National University of Cordoba (UNC), 1:3 for the National University of La Plata (UNLP) and 1:2 for the National University of Rosario (UNR). There were no records of publications from the National Universities of Tucuman and of the North-East. The groups of investigation with greater quantity of presentations and better Publ./Pres. Ratio were Dental Mat./Restorative Dent./Endod. (ratio 1:8), Physiol./Pharmacol./Biochem. (Ratio 1:4) and Oral Pathol. (Ratio 1:4) of UBA, and Physiol./Pharmacol. (Ratio 1:1) of UNC. Most of the publications were in English (86%) and within 4 years of presentation. The current Publ./Pres. Ratios are similar to those communicated in 1990 in the area of the basic investigation and represent acceptable values. An increase in this proportion is expected in the future especially in the area of applied clinical research, since publication would imply prior peer review and thus qualitative evaluation of the work. PMID- 11885084 TI - School nursing services: the early years. AB - This is the first in a series of three articles devoted to school nursing practice past, present, and future. This article highlights development of school health programs in the late 1800s to 1950. Since its inception, school nursing service has paralleled events and developments in society. Nursing services were introduced to treat minor illnesses or injuries of children at school, to provide health education to children in school, and to provide follow-up care and teaching in the home setting in order keep the children healthy enough to attend school. As health care moved away from the home into physician-directed hospitals, nursing services in schools shifted from a focus on the public health nurse to the notion of a nurse-teacher with emphasis on health education. By exploring the practice of the early visionaries in school nursing, one realizes that today's nurses have similar challenges and opportunities in providing quality health care to the school community. PMID- 11885085 TI - Open-air schools. AB - Open-air schools were initiated in 1908 and maintained through the 1930s to treat children with tuberculosis while meeting their educational needs. During this period, treatment of tuberculosis was accomplished in sanitoriums where patients, both adults and children, were isolated and exposed to fresh air and rest. Isolating and institutionalizing children made it difficult for them to obtain schooling. The open-air schools were seen as a method of educating children with tuberculosis while treating them with fresh air, rest, and nutritious food. Although early reports of the effects of open-air schools were positive, student outcome studies reported conflicting results. Some student health outcomes improved, but reports of academic outcomes were inconsistent, leading to their closure between 1938 and 1941. Nursing roles were limited, but nurses were well prepared to participate in the open-air schools by providing health education, maintaining health records, monitoring student health outcomes, and coordinating the services provided to the students. Nurses can learn from our history and hopefully avoid the missed opportunities of yesteryear. PMID- 11885086 TI - Childhood depressive disorders. AB - About 5% of children and adolescents in the general population suffer from depressive disorders at any given point in time. Children under stress, those who experience loss, or children who have attention, learning, conduct, or anxiety disorders are at a higher risk for depressive disorders. Depressive disorders cause diminished quality of life and can ultimately affect mortality through increased risk of suicide and acts of violence. This article describes the incidence and prevalence of childhood depressive disorders, reviews the diagnostic criteria for and the symptom manifestations of depressive illnesses, and discusses recommended treatments. PMID- 11885087 TI - Pediculosis in a school population. AB - This nonexperimental, retrospective study of elementary, middle school, and high school children (a) determined the prevalence of pediculosis, (b) identified populations susceptible to head lice infestation, and (c) examined the role of the school nurse in pediculosis management. The findings showed the highest prevalence of pediculosis was among younger school-age children, and girls were 3 times more likely to be infested than boys. Hispanic children had the highest rate of infestation, followed by Caucasian children. African American children showed a 15% infestation rate, a finding not supported by other studies. The role of the school nurse in pediculosis management was restricted by time and budgetary constraints. Findings of this study support the need for a standardized data collection system for pediculosis in all school districts. PMID- 11885088 TI - Initiation of a school employee wellness program: applying the Comprehensive Health Education Model. AB - Employee wellness is an important component of a coordinated school health program and one that often lacks formalization. With many things competing for the school nurse's attention, health promotion for staff most often consists of helping to arrange flu vaccination and blood pressure screenings. The Comprehensive Health Model assists school personnel desiring to formalize an employee wellness program. An earlier nursing practice management article (Galemore, 2000) explored the history and components of work-site wellness programs. The purpose of this article is to review the process as outlined by the Comprehensive Health Education Model used by one school district to initiate an employee wellness program. PMID- 11885089 TI - Managing risks in professional and clinical performance dilemmas: Part II. AB - The primary purpose of the second article in this 2-part series is to describe and illustrate the use of an analytical framework that may assist school nurses to approach and resolve the dilemmas they may face in practice. Part I of the article was published in the April issue of this journal. It defined the terms "professional performance issue" and "clinical performance issue" and described a 5-step framework for analyzing practice dilemmas related to clinical and performance issues. In this article, the framework will be applied to a specific case scenario involving unsafe staffing and delegation. PMID- 11885091 TI - Transitions: a new look for the journal. PMID- 11885090 TI - An overview of herbal medications with implications for the school nurse. AB - Americans are increasingly using herbs and herbal preparations to treat a wide array of conditions ranging from colds to depression. Children and parents are ingesting herbs in greater quantities, as reflected by increased herb sales and telephone calls to poison information centers. Herbal preparations are being used for the management of insomnia, anxiety, and attention deficit hyperactivity disorders. This trend introduces interesting opportunities and dilemmas for the school nurse. This article provides an overview of the seven most common medicinal herbs (garlic, ginkgo, ginseng, Echinacea, chamomile, valerian, and feverfew). It provides a guide to choosing herbal supplements, a framework for developing school-based herbal policies, and a discussion of the issues faced by school nurses. PMID- 11885092 TI - Early intervention and school nursing practice. AB - The most vital and critical period for early intervention is in the first 3 years of life. The school nurse working in an early childhood program plays a pivotal role in assessing and meeting the health and developmental needs of very young children and their families. This article discusses early intervention, as defined by federal law, as an age-related service delivered within a family focused, multidisciplinary, interagency, and collaborative model. An assessment framework for school nurses working with this population is described, which includes physical, social, and emotional domains. Identifying children early and then providing needed interventions and services will assist at-risk children in realizing their developmental potential. PMID- 11885093 TI - School nursing practice today: implications for the future. AB - School nurses provide health care services to children in a complex environment. School nurses in Delaware established Strategic Planning and Research Committees to assist them in meeting the challenges of providing quality school health care. The small size of the state gave the researcher the opportunity to survey all currently practicing school nurses in public, private, and parochial settings. In this descriptive study, school nurses were asked to identify activities in their daily practice in five areas: administrative, educational, clerical, supportive, and physical. School nurses were asked to rate the frequency at which they performed these activities and to assess their perceived level of competence in the activities. Analysis of the surveys returned identified areas of common practice as well as areas of development and implementation of new school nursing skills. PMID- 11885094 TI - Development and evaluation of a Mexican immigrant family support program. AB - A report to Congress in the fall of 1998 warned that immigrant children are in a state of emergency regarding access to health care. This article presents the three phases involved in developing, implementing, and evaluating a Mexican American Problem Solving (MAPS) program designed to promote the mental health of families. Methods were Phase 1 focus groups to identify concerns and desired approaches for intervention; Phase 2 instrument assessment and prevalence assessment of mental health; and Phase 3 intervention testing. In Phases 1 and 2, 67% of mothers and 59% of children had mental health scores that required referral for evaluation. Participating mothers and children reported positive views of the intervention and showed significant improvements in mental health scores. PMID- 11885095 TI - Computer mentoring for school nurses: the New York State model. AB - As computer use in the school health office is becoming a necessity, learning and upgrading technology skills is a high priority for school nurses across the country. The New York State Association of School Nurses and the Statewide Advocacy for School Health Services conducted a needs assessment to determine school nurses' perceptions of information technology skill level and the use of technology in the school health office. From these data, they have collaboratively developed a computer-mentoring program to be used throughout New York State. This program pairs standardized computer training with the assignment of peer mentors to support novice computer users during the adoption of technology into New York's school health services. PMID- 11885097 TI - A student health advocate program. AB - School nurses are in a unique position to influence health education in the school system. However, it is likely that school nurses have varying opportunity, due to staffing patterns, to effectively use their health education background to reach large numbers of students. This article explores the benefits of providing a school nurse-facilitated student health advocate program. The student health advocate program is a creative approach, designed to access a greater audience of students for health education and health promotion activities. It also cites research supporting the choice of peer education as a health education/health promotion strategy. The initiation of a student health advocate program, the role of the school nurse, student activities, program evaluation, and nursing implications are discussed. Positive benefits accrued for both the student health advocate and the mentored student. PMID- 11885096 TI - The value, be, do ethical decision-making model: balancing students' needs in school nursing. AB - How should school nurses balance the needs of students with complex health problems and the needs of all the other students under their care? School nurses experience this and many other stressful ethical problems. The ethical decision making model, "Value, Be, Do: Guidelines for Resolving Ethical Conflict," provides a philosophical tool for effective resolutions and ethical nursing practice. PMID- 11885098 TI - The future of nursing: responding to the nursing shortage. PMID- 11885100 TI - Articulating your philosophy of nursing. PMID- 11885099 TI - School nursing today: a search for new cheese. AB - The practice of school nursing must change with the times. School nurses must develop new leadership and collaborative skills to work within the interdisciplinary school and community teams that will be required to put changes in place. This article defines current trends and their implications for the search for the most efficient and appropriate school health services. It explores preparation and competencies for school nurses, legal and ethical concerns, and financing needed to provide adequate school health services. Many support systems exist today, and new ones are being developed to expand the range of health services provided in schools. The National Association of School Nurses encourages strategic planning for the new school health environment, and the Office of School Health at the University of Colorado provides extensive resources to help school nurses develop and put their plans in place. PMID- 11885101 TI - Tattooing, body piercing, and branding are on the rise: perspectives for school nurses. AB - This journal presented the first nursing information on adolescents and tattooing 6 years ago, and 5 years ago, information was provided about body piercing. These were published to help school nurses assist adolescents become informed decision makers. Another purpose was to prevent risks and, if possible, help dissuade adolescents from tattooing and body piercing. Continuing this theme, the latest information and trends are reported and discussed, and new information on scarification and branding is presented. If an adolescent wants some form of body art (tattooing, body piercing, or branding), they will often obtain it regardless of regulations, risks, or money. School nurses can take a powerful, proactive role by sharing applicable information, realistic concerns, and care instructions about tattooing, body piercing, and branding. Specific information, risks, and care about each form of procedure is presented. A convenient reference table is available for nurses and students. Additionally, nursing actions are suggested including making changes in health policies regarding body art on a local and state level. PMID- 11885102 TI - Bullying and sexual harassment in the school setting. AB - This article defines bullying and sexual harassment, identifies associated characteristics of the aggressor and the victim, and describes implications for school nurses. The background of federal laws with a focus on the most current 1999 Supreme Court decision, holding a school district liable for damages under federal law (Title IX), is addressed with a case study. Health promotion issues and prevention concepts are outlined in a 10-Point Action Plan to facilitate the prevention and management of bullying and sexual harassment in schools. A survey tool to assess bullying and teaching plans for parents of victims and aggressors are provided. Suggested linkages among approved nursing languages, North American Nursing Diagnosis Association (NANDA) nursing diagnoses (NANDA, 1998), Nursing Interventions Classification interventions (Iowa Intervention Project, 2000), and Nursing Outcomes Classification outcomes (Iowa Outcomes Project, 2000) are included for use in developing nursing care plans for both aggressors and victims of harassment. PMID- 11885103 TI - School health nursing: framework for the future, Part I. AB - Society has an emerging respect for the impact that school health nursing programs have on both the health and the education of school-age children. School nurses need to capture current opportunities by building on the richness of the past in order to remain viable, to advance the value of the specialty, and to meet increasing demands for health-related services in schools. Thriving in an increasingly complex and outcome-driven health and educational environment will require a broadening of perspectives, a modifying of paradigms, and an adjustment of attitudes and practices. This will involve building stronger program support, influencing external societal forces, forging new and expanded partnerships, and solidifying program infrastructures while addressing the specialty's professional and practice issues. Such changes will allow school nursing services to remain available and relevant to the needs of the students and communities they serve. This is Part I of a two-part series on the future of school nursing. PMID- 11885104 TI - Immunization controversy: understanding and addressing public misconceptions and concerns. AB - School nurses often meet with parents who are reluctant to immunize their children. This reluctance is based on widely publicized stories about vaccine safety. Illnesses that are preventable by vaccines have become almost nonexistent, and consequently, vaccine safety concerns have increased in prominence. Often a negative report about the risk of a particular vaccine is released by the media and on the Internet before scientific evidence has been obtained. To adequately respond to parental concerns, school nurses should be aware of the historical impact of vaccine safety issues and the vaccine-related fears that are prevalent at the present time. Nurses also need to be provided with scientifically accurate information so that risk-benefit concerns regarding vaccine safety can be effectively communicated to parents. In this way, school nurses can play an important role in ensuring that the student population and the community are protected from vaccine-preventable diseases. PMID- 11885105 TI - Enhancing wellness in a high school: a community partnership. AB - Meeting the wellness needs of high school students reporting high-risk behaviors above national averages was the purpose of a community partnership between the county school district and West Virginia University School of Nursing. Although the school district and School of Nursing were the primary partners, other programs in the university provided additional support. The school nurse, school of nursing faculty, and nursing students provided wellness programs to students, faculty, and staff. Positive evaluations and high demand for the services demonstrated the school community's need for the program and the success of the partnership. PMID- 11885106 TI - Evaluating the value of screening for hypertension: an evidence-based approach. AB - No recommendations regarding in-school blood pressure (BP) screening currently exist. The purpose of this project was to use an evidence-based approach to determine whether BP screening should be initiated as part of one school district's standard screening protocols. Pediatric BP measurement, risk factors for hypertension, issues for determining youth at risk for hypertension, and eligibility criteria for determining conditions appropriate for screening are discussed. BPs of 1st, 6th, and 11th graders were evaluated according to standardized criteria. The evidence indicated that BP screening in school appears warranted, although a formalized study is needed before a definitive decision can be made regarding the incorporation of BP screening into school health services. PMID- 11885107 TI - "Operation Sick Bay": an opportunity for community service learning. AB - "Operation Sick Bay" was the inspiration of the 3 full-time nurses at Framingham High School, which has a population of 1,800 students and 300 teachers and staff. The primary focus of Operation Sick Bay was to create a health services environment conductive to learning about health and illness, developing lifetime skills in self-help and self-care, providing a restful atmosphere, and involving students and staff in taking pride in their school health services center. The "sick bays" are small units with privacy curtain closures used for students who need to lie down because of illness. There are 3 bays for female students and 3 for male students. Instead of the usual cream-colored walls, the project provided the opportunity to create a colorful environment designed to provide health information to students. PMID- 11885108 TI - New esthetics curriculum--first of its kind. PMID- 11885109 TI - LDS/PDP subsidiary report. Louisiana Dental Services, Inc. PMID- 11885111 TI - Effective communication between dentists and laboratories. PMID- 11885110 TI - HIPAA compliance 101: privacy aspects. PMID- 11885113 TI - Thinking upstream about promoting healthy environments in schools. PMID- 11885112 TI - Second generation anticonvulsant medications: their use in children. AB - The pharmacotherapy of seizure disorders has long relied on a few standard medications such as phenobarbital, phenytoin (Dilantin), valproate (Depakote), and others that represent the "first generation" of anticonvulsants. This article reviews the newer, "second-generation" anticonvulsants that were developed in the last decade. The addition of these second-generation agents has doubled the number of therapies available for the treatment of seizure disorders. They include felbamate (Felbatol), gabapentin (Neurontin), lamotrigine (Lamictal), levetiracetam (Keppra), oxcarbazepine (Trileptal), tiagabine (Gabitril), topiramate (Topamax), and zonisamide (Zonegran). This article describes the known side effects of the second-generation agents and reviews the adverse reactions of the first generation of anticonvulsants as a guide to potential toxicities. Reference tables are included that note usual dosages, available dosage forms, and tablet imprint. In addition, this article describes monitoring parameters and gives specific information regarding the use of these agents. PMID- 11885114 TI - School health nursing: framework for the future. Part II. AB - The relative prestige that school health nursing currently enjoys provides a positive climate in which to advance this nursing specialty. To fully capture this advantage, the profession and its practitioners need to address some practice issues while reinforcing its community-based influence on the health and educational success of America's school-age children. Part I of this series addressed the societal environment in which school nursing finds itself, the factors that support school health nursing programs, and the need to develop new and expanded partnerships. Part II speaks to the professional, practice, and management issues facing school nurses and offers strategies for creating a solid framework in the 21st century. PMID- 11885115 TI - The need for quality physical education. AB - Recently, the U.S. Surgeon General described two aspects of national health that require our immediate attention: As a nation, we are becoming increasingly more sedentary and more overweight. Furthermore, he and some of the most prominent health agencies have suggested that one of the primary strategies to overcome what is viewed as a national epidemic is the promotion of quality physical education in our schools. This paper describes in brief the facts associated with this concern, what is meant by quality physical education, and the role of the school nurse in helping to promote physical education in the schools. PMID- 11885116 TI - School-age caregivers: perceptions of school nurses working in central England. AB - Children are caring for invalid relatives in Britain today, often at the expense of their own development. Research indicates these children are vulnerable to a wide range of problems. A preliminary investigation was conducted on the experiences of school nurses in central England to ascertain what they know about school-age caregivers, their awareness of how caring affects their health, and their perception of support available for caregivers. Eighteen school nurses participated in a qualitative investigation, which indicated that given recognition and resources, school nurses could identify school-age caregivers, their health needs, and, in cooperation with other agencies, provide these children with the necessary support. PMID- 11885117 TI - Acculturation status, birth outcomes, and family planning compliance among Hispanic teens. AB - This study examined acculturation status, selected demographic and pregnancy indices, and the relationship to birth outcomes and family planning patterns among a convenience sample of 63 Hispanic adolescents aged 13 to 19 years and attending community-based prenatal clinics. Findings suggest that Hispanic teenagers who are the first generation in the United States and traditional in their world view are compliant with prenatal and postpartum care and have healthy babies and birth outcomes. Gravidity and gestational age of the infant were significant predictors of birth weight, accounting for 30% of the variability in birth weight. Generation in the United States accounted for 8% of the variance in family planning compliance. Higher gravidity was associated with increased infant birth weight and a decreased likelihood for return for family planning visits during the 1st year postbirth. Teens who were first generation in the United States were more likely to return for family planning visits during the 1st year. School nurses are in a pivotal position to design intervention programs that build on traditional cultural prescriptions for healthy behaviors during and after pregnancy. PMID- 11885118 TI - Statewide demonstration of not on tobacco: a gender-sensitive teen smoking cessation program. AB - This study represented the largest statewide demonstration (n = 346) of the teen smoking cessation program Not On Tobacco (N-O-T) to date and one of the few systematically controlled teen smoking cessation trials reported in the literature. Results showed that N-O-T female teens were 4 times more likely to quit smoking almost 6 months after the program ended than female teens who received a brief intervention (BI). The quit rate for the N-O-T female groups was significantly higher than that for female brief intervention comparison groups. The study demonstrated that 2 times more N-O-T than BI teens quit smoking overall. Differences in the biochemically validated quit rate between the N-O-T groups and the brief intervention groups overall and for male participants were not statistically different, however. Furthermore, findings showed that N-O-T was more effective than the brief intervention in assisting youth with cigarette reduction. There was a significant difference in the reduction rate between the N O-T and the BI groups on weekdays and weekends 6 months after the program ended. Overall, approximately 84% of N-O-T teens either quit or reduced smoking, compared with approximately 55% of BI teens. This study is 1 phase of an ongoing multiphase evaluation of N-O-T. This study resulted in several important findings that will help guide future teen cessation studies and tobacco cessation efforts of school health professionals. PMID- 11885119 TI - Handheld computing: the next technology frontier for school nurses. AB - The purpose of this article is to demonstrate the potential utilization of handheld personal digital assistants for school nurses. Handheld devices and their general uses are described. Clinical programs available through downloading and Web sites with handheld resources are included. Finally, specific handheld computer applications that can be adapted for school nurses are discussed. PMID- 11885120 TI - [Morphologic and electrophysiologic characteristics of cultured vestibular ganglia neurons]. AB - Vestibular afferent neurons have been classified on the basis of their spontaneous activity as regular and irregular; this has been attributed to their synaptic input, but it remains to be defined the participation of some intrinsical properties of the afferent neurons in the determination of their discharge pattern. In this work, we have developed tissue cultures of the rat vestibular ganglia. Isolated cells were plated using poly-D-lysine or collagen as substrates and L-15 or Neurobasal as culture media. After 48 hrs cells in the four experimental conditions give forth neurites of variable longitude. By using antibodies against the neurofilaments 160 kDa the cell structure was studied. Monopolar (30.6%), bipolar (63.9%) and multipolar (5.5%) cells were found. By using the voltage and current clamp procedures the voltage dependence and kinetics of the tetrodotoxin sensitive Na+ current was fully characterized. Cultured cells were shown to generate action potentials under electrical stimulation, and they were capable of repetitive spike discharge under the influence of 4-aminopyridine. These results demonstrate that tissue cultures constitute an excellent system to study the intrinsical properties of vestibular afferent neurons. PMID- 11885121 TI - [Neutropenic colitis]. PMID- 11885122 TI - [Genetic bases of language]. PMID- 11885123 TI - [Controversy between estrogen replacement therapy and risk of breast cancer in menopause]. AB - At present, no information is available from controlled prospective randomized clinical trials to demonstrate a causal link between estrogen replacement therapy (HRT) and the risk of developing breast cancer. In most epidemiologic studies, HRT is not associated with an major increased risk of breast cancer; thus for women who had used estrogen for 10 years or more, the relative risk of breast cancer is 1.46 which is considered as small magnitude. Clinicians and patients are challenged with the difficult task of balancing the beneficial effects of HRT on cardiovascular and bone disease with the potential adverse effects on the breast. The analyses of the benefits and risks of HRT generally indicate that the benefits of therapy outweigh the risks. In other respect the number of survivors of breast cancer are increasing rapidly because of both early detection and the availability of more effective treatments. This effect will increase the number of hypoestrogenic survivors of breast cancer, a group that might benefit from HRT. However, the decision of using HRT has to be determined between the patient and the physician. PMID- 11885124 TI - [Bioethics in genetic engineering]. AB - The advances in the field of molecular biology and genetics have widened the possibilities for the diagnosis and treatment of hereditary diseases. At the same time research into this field has broken bounds of its legal and ethical regulation. The intention of this paper is not to analyze these advances from scientific and technical point of view which is the area of the specialist, but rather to review historical antecedents of genetic engineering; the legal and ethical repercussions of the human genome project (HGP); in vitro fertilization and embryo transfer (FIVET); the embryo research which is being caused out and which may be possible; other fields of genetics and cloning especially germinal cells and human beings; genetic diagnosis and its family, social and work repercussions; treatment through genetic engineering, research into cloning in order to obtain organs and tissues for transplants; and the use of genetic engineering in the biomedical industry. To avoid these advances working against humans, the organization and participation of multidisciplinary bodies are required to provide legal and ethical supervision. PMID- 11885125 TI - [For the patient's autonomy]. PMID- 11885126 TI - [Biomarkers in the prognosis and treatment response of breast cancer]. AB - PURPOSE: To determine the prognostic value of c-erbB-2, p53, hormone receptors and angiogenesis, on recurrence free time and its relationship to treatment in breast cancer patients. METHODS: Women with histologic diagnosis of breast cancer and immunohistochemical determination of biologic factors. Clinic, histologic, molecular factors and recurrence free time were registered. RESULTS: 101 patients, ages 51.98 +/- 11.5 years. Follow-up 32.52 +/- 24.3 months. Tumor recurred in 31, (30.69%); 15 (48.33%) had tumor size above 2.1 cm, 19 (61.29%) showed positive estrogen receptors and 18 (58.07%) for progesterone; 20 (64.51%) to c-erbB-2 expression (64.51%); 18 to p53; average microvessels 24.48 +/- 17.27. Tumor size related to recurrence, p = 0.008. Kruskal-Wallis test did not show a difference when correlating survival free time and biologic factors. 24 pts. (77.41%) received hormones; 20 (64.5%) chemotherapy (61.29%); 19 (61.29%) radiotherapy. Response prediction to hormones with estrogen receptor positive, p = 0.059; to chemotherapy in angiogenesis under 40 vessels/field-0.024. CONCLUSIONS: Tumor size has prognostic implications. A clear positive tendency was observed with p53 and higher microvessel density. Estrogen receptors offer predictive response value to hormone treatment and lower vascular density to chemotherapy, treatment indicators of possible therapeutic association. PMID- 11885127 TI - [Prevalence of bacterial vaginosis in a group of women at a family planning clinic]. AB - OBJECTIVE: To determine the prevalence of bacterial vaginosis among family planning users and the relationship between clinical symptoms and gynecologic signs. MATERIAL AND METHODS: Four hundred and fifty women were studied: they answered a detailed structured questionnaire and had a vaginal secretion sampling to make the diagnose of bacterial vaginosis according to the Amsel et al criteria. RESULTS: 85 women were excluded because they were found to have either yeast or trichomonas. 144/450 women were found to have bacterial vaginosis (32%), while 221 women without evidence of this problem served as a comparison group. We found an increased number of sexual partners, and more abortions and premature rupture of membranes in women with bacterial vaginosis than women without the problem. More than 50% of patients with bacterial vaginosis were asymptomatic of disease complaints and the only sign with significance observed in bacterial vaginosis patients was a grey vaginal discharge. Use of IUD wasn't associated with bacterial vaginosis. CONCLUSIONS: The prevalence of bacterial vaginosis we found in a family planning clinic was similar to those that have been reported in developed countries whereas it seems to be higher than other national studies. We must emphasize the importance of an accurate diagnose and a suitable treatment to prevent subsequent complications. PMID- 11885128 TI - [Vaccination messages foster mobilization and high coverage in Mexico]. AB - OBJECTIVE: To understand the perceptions of the effect of vaccination messages aired during the Second National Health Week in 1996 on the predisposition to vaccinate children among mothers from a lower class neighbourhood in Mexico City. METHODS: 120 mothers of children between the ages of 0 and 7 years who were exposed to the campaign messages participated. They were divided into 8 focus groups based on the age of the children and level of schooling attained by the mothers. A content analysis was conducted using open-ended coding and categorization based on shared concepts. RESULTS: The mothers had a positive image of vaccines for the health of their children. They perceived that the messages reminded them that they needed to vaccinate their children and contributed to the mobilization of their social network in support of vaccination. The authors inferred of mother 5 narratives that some components of messages generated inaccurate interpretations and knowledge and reinforced negative attitudes and cultural and organizational barriers to vaccination in some mothers. CONCLUSIONS: The messages prompted the use of vaccination services during the health campaign. In the production, the form and content should be modified to overcome misunderstandings and barriers to vaccination. PMID- 11885129 TI - [Frequent clinical syndromes without anatomical bases. New perspectives on fibromyalgia and irritable intestine]. PMID- 11885130 TI - [Clostridium difficile infection]. AB - Clostridium difficile is an anaerobic species consisting of bacilli with large, oval, subterminal spores, normally found in intestines. It uses two toxins, which produce cytopathic changes in the intestinal mucosae, causing diarrhea. Patients can present a spectrum of disease that varies from uncomplicated antibiotic-associated diarrhea to life threatening antibiotic-associated pseudomembranous colitis. C. difficile is the only species. There are no defined sterotypes. Toxigenic and nontoxigenic strains exist. The former produce varying amounts of toxin A (enterotoxin) and toxin B (Cytotoxin). Broad spectrum antiboiotic therapy eliminates much competing normal flora, permitting intestinal overgrowth of toxigenic C. difficile. There are no defined host defenses. Metronidazole and vancomycin should be used therapeutically, however, relapses can occur. Supportive therepy may be needed. PMID- 11885132 TI - [A 66-year-old woman with acute respiratory insufficiency, uremic syndrome, abdominal pain, and diarrhea]. PMID- 11885131 TI - [New concepts in the biology of acute myeloid leukemia]. AB - During the last 20 years, several concepts regarding the biology of acute myeloid leukemia (AML) have changed in a profound manner. This has been mainly due to significant advances in the identification, purification and characterization of the primitive hematopoietic cells--including stem and progenitor cells--in which this disorder originates. In the present review article, we discuss some of these new concepts and their relevance in the treatment of AML. PMID- 11885133 TI - [200 years of the smallpox vaccine]. AB - The expression "inoculation of smallpox" was first employed by Emanuele Timone, native of Chios island and graduated from the Universities of Padua and Oxford. He learned about this procedure in Constantinople. This method was introduced in North America, during the great epidemic outbreak of 1721, by two Bostonian citizens: Cotton Mather and Zabdiel Boylston. The French physician Henri Etienne Morel introduced the procedure into New Spain during the smallpox epidemic of 1779. Nevertheless only in 1798 the English physician Edward Jenner published the results of his observations and experience concerning the "vaccination" in his book "Inquiry into the cause and effects of the variolae vaccinae." After some initial oppositions, this method rapidly spreaded to the rest of Europe. It arrived to Spain in 1801 and thence was transferred to Spanish America and Philippine Islands with the expedition leaded by Francisco Xavier Balmis. Along the XIX century the methods for obtaining and keeping the vaccine were notably improved. Both Jenner and Balmis are worthy of remembrance as great humanity benefactors. PMID- 11885134 TI - [Anthology of the first clinical studies with hypothalamic hormones: a story of successful international cooperation]. AB - Our early pioneering clinical trials in Mexico with natural and synthetic thyrotropin-releasing hormone (TRH) and luteinizing hormone releasing hormone (LH RH) also known as gonadotropin releasing hormone (Gn-RH), were reviewed. Highly purified TRH of porcine origin was shown to stimulate Thyrotropin (TSH) release in hypothyroid cretins. Subsequent tests with synthetic TRH also demonstrated significant increases in plasma TSH in normal men and women as well as in patients with primary hypothyroidism and other endocrine disorders. Even more extensive clinical studies were carried out with highly purified natural porcine LH-RH. Subjects with normal basal serum levels of gonadotropins, low levels (men and women pretreated with steroids) and high levels (e.g. post menopausal women) all responded to LH-RH with a release of LH and FSH. The results of these early studies with the natural LH-RH were confirmed by the use of synthetic LH-RH. These investigations made in Mexico with TRH and LH-RH preceded all other clinical studies by a wide margin. Subsequently various clinical investigations with LH-RH agonists and antagonists were also carried out. All these studies played a major role in introducing hypothalamic-releasing hormones into clinical medicine. PMID- 11885135 TI - Pulsed electric field processing of high acid liquid foods: a review. PMID- 11885136 TI - Thermal inactivation of pathogens and verification of adequate cooking in meat and poultry products. PMID- 11885137 TI - Phytoestrogens in foods. PMID- 11885138 TI - Taste and smell perception in the elderly: effect of medications and disease. PMID- 11885139 TI - Structure and mechanical properties of fat crystal networks. PMID- 11885141 TI - Buckwheat: composition, chemistry, and processing. PMID- 11885140 TI - Development and application of multicomponent edible coatings and films: a review. AB - Combining the advantages of polysaccharides, proteins and/or lipids offers multicomponent edible films and coatings good mass transfer barrier properties. Multicomponent edible films and coatings could be beneficial to the food industry by leading to innovative applications. The barrier properties of these systems strongly depend upon their structure and chemistry, the interaction between different film components as well as surrounding environment conditions. Future researches on these multicomponent systems need to concentrate on the following goals: (1) investigation of the optimal film compositions and specific film forming conditions for different food systems; (2) study of the film responses in their barrier properties to environmental factors such as RH and temperature; (3) fundamental research of the mechanism of mass transfer and the interaction of different films components and foods; (4) exploration of the feasibility of potential applications in the food industry. PMID- 11885142 TI - A model of facilitative communication for support of general hospital nurses nursing mentally ill people. Part 2: model description and evaluation. AB - Part 1 of this article dealt with a full description of the research design and methods. This article aims at describing a model of facilitative communication to support general hospital nurses nursing the mentally-ill. In this article a model of facilitative communication applicable to any general hospital setting is proposed. Fundamental assumptions and relationship statements are highlighted and the structure and process of facilitative communication is described according to the three steps employed: 1) assisting the general hospital nurse learn the skill; 2) assisting the general hospital nurse practise the skill in order to develop confidence; and 3) using the skill in a work setting. The guidelines for operationalizing this model are dealt with in the next article. The evaluation of the model is also briefly described. PMID- 11885143 TI - A comparative analysis of ethical development of student nurses registered for a basic degree and basic diploma programme in KwaZulu Natal. AB - A comparative descriptive study was conducted to establish whether the Comprehensive Basic Nursing Course (CBNC) is able to develop students ethically, and how educational preparation from two different programmes (basic degree and basic diploma) influence their ethical development. This study was conducted because of the concerns on the escalating number of litigations instituted against nurses. Several studies have indicated that some of these litigations are as a result of the growing complexity of the health care system and the society's increasing awareness of their human rights. Some studies have shown that nurses are failing to make principled and ethically sound decisions because they are inadequately prepared to handle ethical issues in an ethically responsible manner. A purposively selected sample of third and fourth year students from both programmes was used. Data was collected from both groups through the use of questionnaires. The findings revealed that the students are developing ethically in a CBNC but the level of ethical development is influenced by their educational preparation, teaching approaches and strategies used, clinical environment, hospital bureaucracy, rules and policies. PMID- 11885144 TI - Affairs of the heart: patients' personal constructions of a cardiac event and their effect on lifestyle change. AB - The issue of why people do not always make appropriate lifestyle changes in response to a cardiac event has and continues to be of central importance to health practitioners. This paper addresses this issue from the perspective of the lived experience of persons who have suffered an acute myocardial infarction (AMI). The experiences of 10 persons admitted to the coronary care unit (CCU) of a South African clinic were richly described, making use of the grounded theory methodology. These descriptions were then used as a basis for the development of a contextualist theory of the experience of heart attacks. A central feature of the results was that the disease was mainly attributed to stress by the participants. This was in contrast to the explanations offered by the medical profession, who attribute this more to other modifiable risk factors such as smoking, high blood cholesterol, high blood pressure, and lack of exercise. This tension between lay and professional constructions of the aetiology of the condition is deemed to be of import in the recovery process. The paper further alludes to the goodness of fit which exists between the proposed grounded theory and the personal construct theory of George Kelly. The importance of personal constructions of the event is then used as the basis for a proposed intervention process aimed at addressing the difficulties AMI patients' experience in making and sustaining lifestyle changes. PMID- 11885145 TI - The life world of the adolescent with mental health problems. AB - Adolescents are currently being more and more exposed to the expectations of parents, educators, health-workers/helpers and policy makers to meet the demands of society and conform to it. The perception arises that adults are not able to let the adolescent take responsibility for the HOW of his own life story, despite all the expectations and demands. Under the influence of the post-modernistic approach to science and the narrative therapy it appears that each person is an expert of his own life and that each person is responsible for the how and the writing and rewriting of his own life story. This means that even the adolescent with mental health problems is busy with the writing and rewriting of his life story till even unpleasant incidents and experiences gain new meaning. This demands from the adolescent with mental health problems to be actively involved with his treatment program while the therapist is a participating observer of the therapeutic events. A one-sided approach, where the therapist's objectives and ideas make the difference in the treatment of adolescents with mental health problems, becomes redundant. An alternative approach is suggested where the adolescent with mental health problems becomes co-author of his own life story and his treatment program. In this research the researcher aimed to explore and describe the HOW of the life world of the adolescent with mental health problems. The utilization of the case-study format as research method enabled an in-depth, holistic description of the life world of the adolescent with mental health problems. The implementation of the strategies to ensure trustworthiness, as described by Guba was applied to ensure the validity and reliability of this study. Focus was specifically placed on the application of the strategy of cross validation. This implies that multiple data-collection sources, different experts, theories and respondents were utilized in the exploration of the life world of the adolescent with mental health problems before this life word was described in depth. The researcher makes a few conclusions and based on these make recommendations for application in practice, education and research. PMID- 11885147 TI - A comparison between medicine from an African (Ubuntu) and Western philosophy. AB - I consider the Ubuntu way of caring for the sick in terms of the Ubuntu world view by systematizing the scattered views. I argue that this world-view is underpinned by the regulative concept of sharing and that caring in Ubuntu thinking can only be understood correctly in terms of sharing. I substantiate my exposition in terms of what Africans themselves claim Ubuntu is and relate its meaning to African thinking in general. I consider the uniqueness of this world view by showing how an African thinker compares it to Western World-views on causality and critically consider these comparisons. I apply this world-view to African medicine and evaluate the Ubuntu idea of causes in medicine in comparison with causality in Western thinking by considering the two frameworks of medical care in terms of their viability respectively. I conclude that causal patterns in medicine are controversial in both thinkings but argue that it sets the framework for intercultural communication that can lead both to a better understanding of each other and to some positive developments in medicine. These ways of dealing with the topic represents the significance of this article as an addition to existing knowledge. PMID- 11885146 TI - Knowledge and practice of condom use among first year students at University of the North, South Africa. AB - The aim of the study is to investigate knowledge and sexual practices with reference to correct use of condoms among first year South African University students. The sample consisted of 206 participants, 146 female and 60 male, the mean age was 20.9 years (SD = 3.4), with a range from 17 to 34 years. Results indicated that one third (29.2%) of the sample reported never using condoms, 35.4% always, 19.8% regularly and 8.5% irregularly in the past three months. About 90% levels of correct answers for condom use were found for the items of 'condoms as protection against STD and AIDS', 'expiry date of condoms', and 're using condoms'. More than 15% were not aware that a condom should be put on before any contact with the vagina. The most common mistakes with respect to condom use were ignorance about the correct moment to put on a condom (56%), and when to take off a condom (55%). Male sex and especially increasing recent sexual encounters was associated with correct condom knowledge. The most common reasons for not using a condom were 'I do not have the AIDS virus' and 'I thought I was safe' seems to indicate a low perceived susceptibility. Findings are discussed in view of condom promotion programmes. PMID- 11885148 TI - Student nurses' needs for developing basic study skills. AB - One of the key responsibilities of the nurse educator is to develop student nurses' abilities regarding self-directed study. Self-directed study requires inter alia, the ability to find information, synthesis and consequent application and integration of the information in practice. The development of the abovementioned skills does not only imply a multidimensional approach to the student in totality, but also requires the meticulous involvement of the student in her/his own learning. The latter also assumes that students possess certain essential skills relevant to learning and studying. From the literature it is evident that secondary schooling in general, does not prepare students adequately for tertiary education. This research intended to find answers to the questions whether student nurses require guidance regarding the development of specifically identified study skills, the guidance provided and whether the guidance provided was sufficient. A descriptive survey was done in order to address the above questions. The research instruments (questionnaires) were completed (during 1997) by nurse educators and student nurses in the Western Cape. On completion of the analysis and interpretation of the data, the researcher concluded that student nurses expressed a need for more guidance regarding the development of basic study skills ant that existing student support programs did not address all these needs adequately. Furthermore, it was concluded that the language medium of the prescribed study material had a profound effect on the learning and study processes of student nurses. Based on the conclusion, various recommendations were made concerning different facets of the teaching/learning event., in order to enhance students' learning and studying skills. Mastery of these skills can be regarded as being important prerequisites for effective, responsible, independent professional practice. PMID- 11885149 TI - A model of facilitative communication for the support of general hospital nurses nursing mentally ill people. Part I: background, problem statement and research methodology. AB - The impressive growth in the extent and range of psychiatric services provided by general hospitals in South Africa creates stress among nurses employed in these settings who are not psychiatric trained. This manifests itself in negative attitudes displayed towards mentally ill people. The aim of this paper is to discuss the process followed in the development of the model of facilitative communication. A theory generative design was used. The research methods were dealt with in four steps of theory generation as set out below. Step 1 entailed concept analysis. This step was dealt with in two phases, namely concept identification and concept definition. During concept identification, a qualitative research strategy that is explorative, descriptive and contextual was used. This was achieved through field research conducted in an urban general hospital. A sample of twelve professional nurses was selected from a population of 800 professional nurses employed in a general hospital using the purposive sampling technique. This sample size was determined by saturation of data in themes. Both semi-structured individual phenomenological interviews and observations were used as methods of data collection. Giorgi's method of descriptive data analysis (1985) was used. Four themes emerged from the results of the study. The main concepts of the model were identified and classified using a survey list of Dickoff et al. (1968). Step 2 dealt with the creation of interrelationship statements between concepts identified in Step 1, while Step 3 dealt with the description of the model using strategies proposed by Chinn and Kramer (1991). In Step 4, the description of guidelines for operationalizing in practice was ensured. To ensure valid results, a model for trustworthiness proposed by Guba (Lincoln & Guba, 1985) was used. The following criteria for trustworthiness were applied in all the steps of theory generation: truth value, applicability, consistency and neutrality. PMID- 11885151 TI - The role of the enrolled nursing auxiliary in a selected health care administration. AB - The purpose of the study was to determine the contribution of nursing auxiliaries towards health care services, against their scope of practice. The sample consisted of nursing auxiliaries in the Gazankulu area, in the Northern Transvaal. The findings revealed that nursing auxiliaries are presently an essential component of nursing services rendered in Gazankulu, but that apart from their prescribed role, they are also engaged in activities which should be performed by enrolled and professional nurses, and general assistants. A need for education is apparent for all categories of nursing staff regarding the scope of practice of nursing auxiliaries. PMID- 11885150 TI - The effect of type 2 diabetes mellitus on health-related quality of life (HRQOL). AB - Improving the quality of life of all South Africans has become a major concern to health care practitioners, organisations and politicians. However, the paucity of local information on health-related quality of life (HRQOL) does not allow us to address this public health challenge. In order to rectify this deficiency and complement international research, we undertook a study with 281 Type 2 Black diabetic patients and 437 controls, with no self-reported chronic conditions, to ascertain HRQOL. We used the SF-20 to measure functioning, general health, well being and bodily pain (HRQOL). It was hypothesised that diabetes mellitus significantly affects functioning, general health and well-being. Multiple analyses of covariance controlled for age, schooling, marital status, employment status and commodity ownership (a socio-economic measure). Patients were significantly more likely to report poorer role functioning, poorer general health and more pain than controls, providing partial support for the hypothesis. Reliability (internal consistency) coefficients on the four multi-item SF-20 sub scales ranged between 0.79 (well-being), 0.81 (general health), 0.83 (physical functioning) and 0.94 (role functioning) for patients: for controls these coefficients ranged between 0.70 (well-being), 0.78 (general health), 0.80 (physical functioning) and 0.90 (role functioning). Inter-correlations among the sub-scales were significant for patients and controls (p = 0.01). It was concluded that the SF-20 is a reliable instrument for measuring HRQOL in both patient and control samples, and diabetes mellitus has more impact on general health and level of pain than on well-being. PMID- 11885152 TI - Nurse educators' perceptions of OSCE as a clinical evaluation method. AB - The South African Qualification Authority, and the South African Nursing Council are in pursuit of quality nursing education to enable the learners to practise as independent and autonomous practitioners. The educational programme should focus on the facilitation of critical and reflective thinking skills that will help the learner to make rational decisions and solve problems. A way of achieving this level of functioning is the use of assessment and evaluation methods that measure the learners' clinical competence holistically. This article is focused on the perceptions of twenty nurse educators, purposively selected from three Nursing Colleges affiliated to a university in Gauteng, regarding the use of OSCE (Objective Structured Clinical Examination) as a clinical evaluation method within a qualitative and descriptive research strategy. Three focus group interviews were conducted in different sessions. A descriptive content analysis was used. Trustworthiness was ensured by using Lincoln and Guba's model (1985). The results revealed both positive and negative aspects of OSCE as a clinical evaluation method with regard to: administrative aspects; evaluators; learners; procedures/instruments and evaluation. The conclusion drawn from the related findings is that OSCE does not measure the learners' clinical competence holistically. It is therefore recommended that the identified negative perception be taken as challenges faced by nurse educators and that the positive aspects be strengthened. One way of meeting these recommendations is the use of varied alternative methods for clinical assessment and evaluation that focus on the holistic measurement of the learners' clinical competence. PMID- 11885153 TI - The journey of recovery after a rape experience. AB - In this existential-phenomenological investigation seven women were interviewed about their experiences of recovering from rape trauma. The purpose of the study was to discover the meaning of recovery from the perception of the victim, how recovery is experienced, and what contributed to the growth and recovery of the woman who has been raped. Transcribed interviews were analyzed using a hermeneutic process. The thematic structure of a woman's recovery from rape comprises three main themes: reaching out, reframing the rape, and redefining the self. These findings are important to professionals working with women who have been raped because it is the raped woman, rather than the clinician, who is able to define what constitutes recovery. PMID- 11885154 TI - Dynamically fluctuating hope, despair and hopelessness along the HIV/AIDS continuum as described by caregivers in voluntary organizations in Finland. AB - The purpose of this study was to describe the dynamics of hope in a) people fearing a diagnosis of HIV or living with HIV/AIDS and b) their significant others, from the perspective of caregivers working in voluntary organizations in Finland. Individual interviews with eight caregivers were analysed using the grounded theory method. Living with the fluctuating waves of hope, despair, and hopelessness based on factors constructing them emerged as the core category describing the dynamics of hope in a person fearing a diagnosis of HIV, becoming aware of HIV contagion, and living with HIV/AIDS. Mirroring the fluctuating waves of hope, despair, and hopelessness based on factors constructing them emerged as the core category describing the dynamics of hope in a significant other of a person fearing a diagnosis of HIV, or living with HIV/AIDS. It is important to take into consideration the dynamics of hope in taking care of people fearing a diagnosis of HIV or living with HIV/AIDS and their significant others. PMID- 11885155 TI - A profile of who completes and who drops out of domestic violence rehabilitation. AB - Despite changes made in domestic violence (DV) programs, attrition continues to be a major problem. For this study on DV rehabilitation attrition, 62 male batterers and 31 female victims were recruited during a six month time frame from an existing batterers' program. Of the 62 batterers, one man was removed from the study, 38 dropped out of the program, and 23 made the transition from rehabilitation to the maintenance phase of the program. A logistical regression to predict completion status resulted in a Model Chi-square statistic of 31.08 (p = .000). Completers were more likely young, court-monitored, had lower levels of stress (SOS Inventory) and posttraumatic stress (PCL), and had higher levels of mutuality (MPDQ) in their relationships than noncompleters. The model predicted 88.89% of the noncompleters, 78.26% of the completers, and had an overall predictive ability of 84.75% for the study sample. PMID- 11885156 TI - The meaning of depression from the life-world perspective of elderly women. AB - Depression is a serious public health problem that particularly affects women and elderly people. The aim of this phenomenological study was to gain a deeper understanding of depression in elderly women by investigating and describing the meaning of depression from a life-world perspective. Qualitative interviews were conducted with five elderly women suffering from depression; they were transcribed and analysed using the phenomenological method. The essence of depression emerged as 'reexperiencing a severe personal insult' and the perception of 'increased sensitivity and vulnerability.' These two components constituted the breeding ground for an additional five characteristics. Depression was perceived as a severe multidimensional suffering that affected physical, mental, social, and spiritual aspects, where previous experiences merged with the current situation. Thus, the whole life space of these women was affected. In its most severe form, depression and, thereby, life was perceived as unbearable. The meaning that emerged from the women's descriptions can be understood against the background of the aging individual's retrospection and summation of life. PMID- 11885157 TI - Gender and mental illness: an Australian overview. AB - This article highlights the centrality of gender to mental health nursing practice by providing evidence that gendered assumptions are embedded in psychiatric knowledge. After a brief account of gendered rates of mental illness, the first two-thirds of this article explores formal psychiatric diagnostic criteria, casebook specificity, and processes involved in gaining a psychiatric diagnosis in relation to gender. In contemporary psychiatric practice the two tendencies of overdiagnosis and underdiagnosis are simultaneously evident, with woman-predominant styles of expressing distress being particularly associated with underdiagnosis. The final sections of the article outline gendered attitudes and expectations that impact on clients and their treatment, along with common gendered differences relevant to mental health nursing and people living with anxiety, depression, substance abuse, psychosis, and dual diagnosis. PMID- 11885158 TI - Effects of aggressive behavior and perceived self-efficacy on burnout among staff of homes for the elderly. AB - This study elicits effects of experienced aggressive behavior and perceived self efficacy in coping with aggressive behavior on the dimensions of burnout of staff caring for the elderly (N = 551). From the results of the hierarchical regression analysis it appears that physical and psychological aggression and the number of weekly working hours has an effect on emotional exhaustion of staff. Psychological aggression is found to have an effect on depersonalization. The number of weekly working hours and the perceived self-efficacy in turn appear to have an effect on personal accomplishment. Neither sex nor age has an effect on the burnout dimensions. Implications for research and suggestions for work and training of staff caring for the elderly are discussed. PMID- 11885159 TI - High-risk behavior in teens: self-destructive or adaptive? PMID- 11885160 TI - Evaluation of a youth tobacco education program: student, teacher, and presenter perspectives. AB - Few published studies have explored the impact of smoking prevention programs among elementary-school children. This study describes a qualitative, cross sectional evaluation of the Tar Wars tobacco prevention program among 5th-grade students (n = 888), along with impressions from classroom teachers and program presenters. Results from this evaluation reveal that all constituencies involved with the Tar Wars program--5th-grade students, classroom teachers, and program presenters--indicated high satisfaction with this youth tobacco education program. Students enjoyed the program and indicated understanding of key themes, classroom teachers stated that the program was worthwhile in presenting unique information, and presenters were enthusiastic about the ease of presentation and opportunities for future presentations. PMID- 11885161 TI - A school-based intervention program to prevent adolescent smoking. AB - Research has shown that tobacco use usually begins in early adolescence, results in an increase in future health problems, and ultimately affects national health care costs. Despite the messages about the dangers of smoking, young people continue to smoke. A school-based tobacco education program designed to produce a more favorable attitude about the positive effects of not smoking and increase knowledge of the hazards of smoking was implemented for 6th graders in a parochial middle school. After the intervention, there was a significant increase in knowledge about tobacco but no change in attitudes regarding the use of tobacco. The results have implications for school nurses who design and teach programs to prevent tobacco use. PMID- 11885162 TI - Cigar and marijuana use: their relationship in teens. AB - There is evidence of a substantial increase in teen use of both cigars and marijuana over the last decade. The theoretical concept of sensation seeking suggests a conceptual relationship between teen use of the substances. The hypothesis was that a relationship existed between the use of cigars and marijuana. Data for this study were gathered during the evaluation in a local school district of a teen tobacco-use reduction program. Questionnaire data and focus group results supported the study hypothesis and provided the direction for peer tobacco education to include an emphasis on this significant relationship. Identified associations between use of different substances supplies cues for the school nurse and other health providers to pursue specific areas of substance use practices when assessing students. PMID- 11885163 TI - Smoking cessation programs for adolescents. AB - Many smoking cessation programs for adult smokers are described in the medical and nursing literature. Programs designed to prevent smoking among adolescents and children also are prevalent in the literature. Despite these prevention programs, many adolescents choose to start smoking. Once adolescents begin smoking, it is difficult to find ways to help them quit. Few programs exist that are targeted to help this population with smoking cessation. This article provides an in-depth review of 5 smoking cessation programs designed for adolescents. Each of the programs presents unique strategies for helping teenage smokers quit. Techniques used in these programs include peer leadership, nicotine patch therapy, peer support, computer instruction, and one-on-one counseling with a nurse practitioner. Each program was studied for efficacy with adolescents. Although none of the programs reviewed showed remarkable success, they serve as guides for future program development. Additional programs need to be developed and studied with larger, more diverse populations. Nurses must identify or develop smoking cessation programs that meet the needs of all types of adolescents and are effective in helping them to quit. Once designed, these smoking cessation programs should be made accessible to adolescents in a variety of settings. PMID- 11885164 TI - Power up your presentation with PowerPoint. AB - School nurses have the experience, insight, and passion to speak in pubic forums on issues regarding the health and education of children. They are seeking opportunities to inform district decision makers and the general public about student needs and about the importance of having adequate resources to meet these needs. Nurses may be called upon to speak both formally and informally to parent and community groups, the school board, and to their peers. To be credible and to communicate their messages effectively, nurses must hone their presentation and speaking skills, which may include the use of up-to-date technology. PMID- 11885165 TI - National school nurse certification. Part I: An ongoing process. AB - The National Association of School Nurses proposed the concept of certification for school nurses in the 1970s. The development and evolution of the school nurse certification process, from concept to reality, is described in this article. It is the first of a 2-part series on national certification for school nurses. Part 2 will examine issues and answer questions about the certification, give information from past and current presidents and certified school nurses about their experiences, and present a forecast about the future of school nurse certification. PMID- 11885166 TI - School health services after Cedar Rapids Independent School District v. Garret F. AB - In 1999, the U.S. Supreme Court ruled that "school health services" include nursing services that are required for students needing one-on-one ("private duty") nursing care to attend school. This ruling settled opposing federal court interpretations of the Individuals with Disabilities Education Act, clarifying the rights of students with complex medical needs and the responsibility of school districts to provide them with an appropriate educational program. In responding to the challenges and opportunities raised by this decision, school nurses must understand the implications for school nursing practice and for school districts, educate their school leaders and communities, and provide leadership for creative and collaborative problem solving. PMID- 11885167 TI - Introduction, methodology, and summary of results for the Dental Implant Clinical Research Group studies. PMID- 11885168 TI - Survival of various implant-supported prosthesis designs following 36 months of clinical function. AB - BACKGROUND: The use of endosseous dental implants to replace natural teeth lost to trauma, dental caries, or periodontal disease has become a predictable form of prosthetic treatment since gaining popularity in the early 1980s. While numerous clinical studies have focused on the survival of implants, few address the survival of different prosthesis designs. METHODS: Beginning in 1991, 882 prostheses supported by more than 2,900 implants (687 patients) were placed by the Department of Veterans Affairs Dental Implant Clinical Research Group (DICRG). These prostheses were divided into five research strata based on arch location. The recommended design for each stratum was: bar-supported overdenture (maxillary completely edentulous); screw-retained hybrid denture (mandibular completely edentulous); screw-retained fixed partial denture (mandibular and maxillary posterior partially edentulous); and cemented single crown (maxillary anterior single tooth). Alternative overdenture designs were utilized in the edentulous arches when the recommended prosthesis could not be fabricated. Prosthesis success rates for the research strata were calculated for an observation time of up to 36 months following prosthesis placement. RESULTS: Success rates for the maxillary edentulous stratum ranged from 94.6% for the bar retained overdenture supported by five to six fixtures to 81.8% for the cap retained overdenture. The mandibular edentulous strata produced success rates of 98.1% for the fixed hybrid prosthesis to 91.7% for the cap-retained prosthesis. Success rates for maxillary and mandibular posterior fixed partial dentures were 94.3% and 92.6%, respectively, while the maxillary anterior single-tooth prosthesis yielded a success rate of 98.1% for the 36-month observation period. CONCLUSIONS: The recommended prosthesis designs investigated in this study proved to be reliable, with encouraging success rates for an observation period of 36 months following placement. PMID- 11885169 TI - Implant surface coating and bone quality-related survival outcomes through 36 months post-placement of root-form endosseous dental implants. AB - Survival rates from placement to 36 months were reported for the ongoing Dental Implant Clinical Research Group studies of root-form endosseous dental implants. Failure rates for all implants were similar in bone qualities 1 and 2 (6.2% and 6.7%, respectively) and slightly higher in bone qualities 3 and 4 (8.5% and 8.7%, respectively). Hydroxyapatite (HA)-coated implants had an overall failure rate of 3.9% over 36 months in all bone qualities combined, while non-coated implants had a 13.4% failure rate for the same parameters. For each bone quality, there was a significant difference in implant survival for the non-coated implants (P < 0.01). The highest failure rates for non-coated implants were in bone qualities 3 and 4 (19.1% and 25.5%, respectively). No major difference in survival was found for HA-coated implants placed in each bone quality. Possible reasons for the differences in survival are discussed. PMID- 11885170 TI - The influence of bone thickness on facial marginal bone response: stage 1 placement through stage 2 uncovering. AB - BACKGROUND: Various causes of facial bone loss around dental implants are reported in the literature; however, reports on the influence of residual facial bone thickness on the facial bone response (loss or gain) have not been published. This study measured changes in vertical dimension of facial bone between implant insertion and uncovering and compared these changes to facial bone thickness for more than 3,000 hydroxyapatite (HA)-coated and non-HA-coated root-form dental implants. METHODS: Subjects were predominantly white males, 18 to 80+ years of age (mean 62.9 years), who were patients at 30 Department of Veterans Affairs Medical Centers and two university dental clinics. Alveolar ridges ranged from normal to resorbed with intact basal bone. Following preparation of the osteotomy site, direct measurements with calipers were made of the residual facial bone thickness, approximately 0.5 mm below the crest of the bone. The distance from the top of the implants to the crest of the facial bone was also measured using periodontal probes. Implants were uncovered between 3 to 4 months in the mandible and 6 to 8 months in the maxilla after insertion. Facial bone response was the difference between the height of facial bone at Stage 1 (insertion) and Stage 2 (uncovering). RESULTS: The mean facial bone thickness after osteotomies were made was 1.7 +/- 1.13 mm. When a mean facial bone thickness of 1.8 +/- 1.41 mm or larger remained after site preparation, bone apposition was more likely to occur. The mean facial bone response for 2,685 implants was -0.7 +/- 1.70 mm. For implants integrated at uncovering, the mean bone response was -0.7 +/- 1.69 mm, and -2.8 +/- 1.57 mm for implants mobile at uncovering. Bone quality-4 had the least facial bone response, -0.5 +/- 2.11 mm. Bone responses were similar for both HA-coated and non-HA-coated implants. CONCLUSIONS: Significantly greater amounts of facial bone loss were associated with implants that failed to integrate. As the bone thickness approached 1.8 to 2 mm, bone loss decreased significantly and some evidence of bone gain was seen. There was no statistically or clinically significant difference in bone response between HA-coated and non-HA-coated implants. PMID- 11885171 TI - The Dental Implant Clinical Research Group study: study design and statistical methods utilized. PMID- 11885172 TI - Influence of research center on overall survival outcomes at each phase of treatment. AB - BACKGROUND: Clinical studies of dental implants tend to fall into two broad categories. Efficacy studies apply strict exclusion criteria under carefully controlled conditions to produce a narrow range of results. Effectiveness studies more closely model real-world treatment environments, with a more diverse patient sample and broader range of provider skills. In this multi-center study of more than 2,900 dental implants, study centers were grouped by implant survival scores in an attempt to draw attention to the influence of confounding variables associated with the treatment environment. METHODS: Thirty-two study centers were ranked by implant survival scores at uncovering and assigned to three performance groups. Centers whose overall scores were within approximately one standard deviation of the mean were placed in the middle (70%) performance group (MPG). The remaining centers were placed in either the top (15%) performance group (TPG) or the lower (15%) performance group (LPG). Overall survival and survival by phase of treatment were recorded for each of six implant designs in each of the three performance groups. RESULTS: From implant placement to 36 months, the TPG achieved survival rates from 100% (for 3 designs) to 95.5% (for one design), with an average of 97% for all designs. Increased variations in survival (97.2% to 73%) occurred in the MPG, with larger variations (96.4% to 48%) in the LPG. The HA-coated cylinder recorded consistently high survival scores (over 95%) in all performance groups and all phases of treatment. Failures for other designs in the MPG and LPG were concentrated in the healing period (placement to uncovering), except for the commercially pure titanium screw, which had the most failures between uncovering and prosthesis loading. CONCLUSIONS: Implant design and treatment environment both play an important role in implant survival. Two design characteristics appear to enhance survival: 1) a surgical protocol involving minimal instrumentation at placement, and 2) hydroxyapatite (HA) coating. The HA coated press-fit cylinder design was the least affected by the center's performance. PMID- 11885174 TI - Survival and stability (PTVs) of six implant designs from placement to 36 months. AB - BACKGROUND: Numerous new implant designs and materials have become available over the last decade, each with special claims of superiority in restoring complex cases. Differences in existing clinical databases, study designs, and methods of determining failures/survival are seldom standardized, which complicates comparisons of clinical performance of these new designs. Little information is available concerning the changes in stability of various designs and materials following clinical loading. METHODS: A total of 30 VA medical centers and 2 dental schools combined to form the Dental Implant Clinical Research Group (DICRG). More than 2,900 implants were placed, restored and data gathered from the time of placement to 36 months. Implant stability from uncovering to 36 months and survival from placement to 36 months were determined. Survival was determined using two different approaches--considering all implants removed at any time, regardless of the reason (DICRG approach), and considering only those that were removed following loading of the prosthesis (post-loading approach). Survival was also determined for each of the three phases of implant treatment- phase 1, from the time of placement to uncovering and abutment connection; phase 2, from uncovering to placement and loading of the prosthesis; and phase 3, from loading of the prosthesis to 36 months. RESULTS: The two approaches to determining survival for each implant design and/or material included in the study showed differences in reported numbers ranging from 1.1% to 21.7%. The largest difference in survival was for the commercially pure titanium screw (used in this study only in the maxillary completely edentulous applications), which showed a 21.7% greater survival rate. With the DICRG approach, the hydroxyapatite (HA)-cylinder had the highest survival (97.5%). When considering the post-loading approach, the titanium-alloy screw had the highest survival (99.4%), with the HA cylinder having the next highest survival (98.6%). The HA-cylinder did not show increased stability from uncovering to 36 months, and the HA-grooved implant became less stable. CONCLUSIONS: HA-coated implants demonstrated the highest survival rate; 2) the post-loading analysis approach inflated survival; 3) non-HA implants showed increased stability following loading; 4) HA-coated implants showed a slight decrease or no change in stability; and 5) the clinical significance of the changes in implant stability must be determined for the long term. PMID- 11885173 TI - Factors associated with radiographic vertical bone loss around implants placed in a clinical study. AB - The loss of vertical bone height over time has been assessed radiographically as part of the Dental Implant Clinical Research Group studies. Radiographs were assessed from implant placement, uncovering surgeries, and recall appointments. Overall, the study implants experienced most peri-implant vertical bone loss in the first year after placement, followed by a dramatic decrease in bone loss rate through the subsequent study intervals. Stratified analysis of data up to 72 months after implant uncovering indicates different bone loss patterns by: 1) arch; 2) jaw region; 3) case type; 4) bone quality; 5) surface type; 6) implant design; 7) smoking status; and 8) postoperative antibiotic treatment. These results will be used to build statistical mixed models to indicate which clinical factors are most predictive of peri-implant vertical bone loss, controlling for confounding and accounting for correlation of data over time and within study patients. PMID- 11885175 TI - Long-term assessment (5 to 71 months) of endosseous dental implants placed in the augmented maxillary sinus. AB - BACKGROUND: It is not uncommon for the placement of endosseous dental implants in the maxillary posterior jaw region to be complicated by the pneumatization of the maxillary sinus. When this occurs, the residual bone between the floor of the sinus and the crestal ridge is inadequate for the placement of implants. The sinus lift procedure provides a way to increase the amount of available bone and the placement of longer implants. METHODS: One hundred twenty (120) implants were placed in 45 augmented maxillary sinuses. Patients ranged in age from 34 to 78 years. The implant design included a limited number of non-hydroxyapatite (HA) coated titanium screws, with the majority of the implants being HA-coated cylinders, grooved cylinders, and screws. The augmentation materials were autogenous bone, allogenic bone (demineralized freeze-dried bone allograft, DFDBA), alloplastic bone (HA), combination grafts of HA and DFDBA, and combination grafts of autogenous bone and DFDBA. All the cases were successfully restored with implant-supported, bar-retained overdentures or fixed partial dentures. The follow-up began at Stage 2 uncovering and ranged from 5 to 71 months, with a mean of 38.2 and standard deviation of 14.6 months. RESULTS: Three (2.5%) of the 120 implants failed between the period of implant placement and 36 months. Failures appeared to be associated with a history of smoking. Other complications encountered during the study are presented. Implant survival was higher in those placed in grafted sinuses (97.5%) than in those placed in the posterior maxilla without sinus grafting (90.3%). CONCLUSION: These findings support the use of implants placed in augmented sinuses to support dental prostheses. PMID- 11885176 TI - Implant survival in patients with type 2 diabetes: placement to 36 months. AB - BACKGROUND: Because the life expectancy of individuals continues to increase, dentists providing dental implant treatment can expect to see an increasing number of patients with diabetes mellitus. Today, there are little data available concerning the clinical outcomes involving the use of implant treatment for patients with diabetes mellitus. There are three types of diabetes mellitus: Type 1 (insulin dependent), Type 2 (non-insulin dependent), and gestational. Because of possible complications from patients with diabetes mellitus, they are excluded from participation in most clinical studies of endosseous dental implant survival. METHODS: This study attempted to determine if Type 2 diabetes represents a significant risk factor to the long-term clinical performance of dental implants, using the comprehensive DICRG database. Diabetes was a possible exclusion criterion; however, the final decision on Type 2 patients was left to the dental implant team at the research center. A total of 2,887 implants (663 patients) were surgically placed, restored, and followed for a period of 36 months. Of these, 2,632 (91%) implants were placed in non-diabetic patients and 255 (8.8%) in Type 2 patients. Failures (survival) were compared using descriptive data. Possible clustering was also studied. RESULTS: A model assuming independence showed that implants in Type 2 patients have significantly more failures (P = 0.020). However, if correlations among implants within the patient are considered, the significance level becomes marginal (P = 0.046). The experience of the surgeon did not produce a clinically significant improvement in implant survival. The use of chlorhexidine rinses following implant placement resulted in a slight improvement (2.5%) in survival in non-Type 2 patients and a greater improvement in Type 2 patients (9.1%); the use of preoperative antibiotics improved survival by 4.5% in non-Type 2 patients and 10.5% in Type 2 patients. The use of HA-coated implants improved survival by 13.2% in Type 2 diabetics. CONCLUSION: Type 2 diabetic patients tend to have more failures than non-diabetic patients; however, the influence was marginally significant. These findings need to be confirmed by other scientific clinical studies with a larger Type 2 diabetic sample size. PMID- 11885177 TI - The influence of preoperative antibiotics on success of endosseous implants at 36 months. AB - The benefits of prophylactic antibiotics are well recognized in dentistry. However, their routine use in the placement of endosseous dental implants remains controversial. As part of the comprehensive Dental Implant Clinical Research Group (DICRG) clinical implant study, the preoperative or postoperative use of antibiotics, the type used, and the duration of coverage were left to the discretion of the surgeon. These data for 2,973 implants were recorded and correlated with failure of osseointegration during healing (Stage 1), at surgical uncovering (Stage 2), before loading the prosthesis (Stage 3), and from prosthesis loading to 36 months (Stage 4). The results showed a significantly higher survival rate at each stage of treatment in patients who had received preoperative antibiotics. PMID- 11885178 TI - A methodological study for the analysis of apatite-coated dental implants retrieved from humans. AB - The stability of thermally processed hydroxyapatite coatings for oral and orthopedic bioprostheses has been questioned. Information on the chemical changes, which occur with hydroxyapatite biomaterials post-implantation in humans, is lacking. The purpose of this investigation was to begin to examine post-implantation surface changes of hydroxyapatite-coated implants using scanning electron microscopy (SEM), x-ray microanalysis (EDAX), Fourier transform infrared spectroscopy (FTIR), and x-ray diffraction (XRD). Three retrieved dental implant specimens from humans following clinical failure due to peri-implantitis were examined. Unimplanted cylinders served as controls. Clinically, the retrieved specimens were all enveloped by a fibrous tissue capsule with bone present at the apical extent of the implant. SEM analysis showed that the retrieved surfaces were coated with both calcified and proteinaceous deposits. EDAX scans of the retrieved specimens demonstrated evidence of hydroxyapatite coating loss reflected by increasing titanium and aluminum signals. Other foreign ions such as sodium, chloride, sulfur, silica, and magnesium were detected. XRD of the control specimens showed that the samples were predominantly apatite; however, two peaks were detected in the diffraction pattern, which are not characteristic of hydroxyapatite, indicating that small amounts of one or more other crystalline phases were also present. The retrieved specimens showed slightly larger average crystal size relative to the control sample material, and the non-apatite lines were not present. FTIR evaluation of the retrieved specimens revealed the incorporation of carbonate and organic matrix on or into the hydroxyapatite. Narrowing of and increased detail in the phosphate peaks indicated an increase in average crystal size and/or perfection relative to the controls, as did the XRD results. Based on these results, we conclude that chemical changes may occur within the coating, with the incorporation of carbonate and concomitant reduction in hydroxyapatite coating thickness. Thermodynamic dissolution-reprecipitation of the coating itself and subsequent surface insult by bacterial and local inflammatory components may be involved with these changes. PMID- 11885179 TI - Implant survival to 36 months as related to length and diameter. AB - BACKGROUND: It is generally accepted that diameter and length of an endosseous dental implant and its stability at placement are critical factors in achieving and maintaining osseointegration. In the event of slight implant mobility at placement, the conventional or accepted treatment is to place a longer implant and/or one of wider diameter. This manuscript presents stability and survival/failure data for implants of different diameters and lengths following 36 months post-placement, as well as crestal bone loss data between placement and uncovering. METHODS: A subset of the Dental Implant Clinical Research Group's database was used to study the 3-year survival and stability of various implant lengths (7 mm, 8 mm, 10 mm, 13 mm, and 16 mm) and diameters (3 mm+ and 4 mm+). Placement to uncovering crestal bone loss was also determined. The implants were generally representative of those available for clinical use (screws, basket, grooved, hydroxy-apatite-coated, CP-Ti, Ti-alloy). The study protocol specified that the implants be randomized to various jaw regions to accomplish the primary goals of the study--the comparison of each implant design's overall survival. A total of 2,917 implants were placed, restored, and followed. Data for all 3 mm to 3.9 mm diameter implants were pooled into a "3+" group, and the 4 mm to 4.9 mm diameter implants into a "4+" mm group. No attempt was made to look at the influence of any other variables on survival outcomes. The possible influence of clustering on survival was taken into consideration. RESULTS: The 3+ mm group had a mean stability (PTV) of -3.8 (SD = 2.9), and the 4+ group had a mean PTV of 4.4 (SD = 2.7) (P < 0.05). The PTVs for implant lengths ranged from -2.9 (SD = 2.8) for 7 mm lengths to -3.9 (SD = 2.9) for 16 mm lengths (P < 0.05). Survival to 36 months was 90.7% for the 3+ diameter and 94.6% for the 4+ group (P = 0.01). Survival ranged from 66.7% for the 7 mm implants to 96.4% for 16 mm implants (P = 0.001). Outcomes did not change when clustering was considered, although the P value decreased slightly. CONCLUSIONS: The results indicate that: 1) shorter implants had statistically lower survival rates as compared with longer implants; 2) 3+ mm diameter implants had a lower survival rate as compared with 4+ mm implants; 3) 3+ mm diameter implants are less stable (more positive PTVs) than 4+ mm implants; and 4) there was no significant difference in crestal bone loss for the two different implant diameters between placement and uncovering. PMID- 11885180 TI - Three-year post-placement survival of implants mobile at placement. AB - BACKGROUND: Although rigid fixation of endosseous implants at the time of placement is generally thought to be a prerequisite for successful osseointegration, the Dental implant Clinical Research Group (DICRG) of the Department of Veterans Affairs has reported on implants that integrated despite being mobile at placement. The present study examines the frequency of osseointegration and the 36-month post-placement survival of implants mobile at placement in a prospective, multicenter, longitudinal clinical study of more than 3,000 implants conducted by the DICRG. METHODS: A total of 3,111 implants of 6 different designs were placed in all jaw regions in more than 800 patients at 32 study centers. At the time of this report, 2,770 of these implants had been followed for 36 months post-placement. They included 89 implants that were mobile at placement. Data for demographic variables, implant coating, bone quality, incision type, bone augmentation, and antibiotic usage were recorded. An electronic hand-held probe was used to measure mobility at uncovering and at regular follow-up intervals. RESULTS: Eighty-nine of 2,770 inserted implants were mobile at placement. Results are reported for two periods: from placement to 36 months and from prosthetic loading to 36 months. The latter method eliminated early failures and resulted in substantially higher scores for both mobile implants at placement (95.9% survival from prosthetic loading to 36 months post placement versus 79.8% from placement to 36 months) and implants not mobile at placement (98.4% versus 93.4%). Mobility at placement was significant to 3-year survival (P < 0.001). Hydroxyapatite (HA) coating improved the performance of implants mobile at placement (91.8% for HA-coated versus 53.6% for non-HA) and those not mobile at placement (97.2% for HA-coated versus 87.4% for non-HA). Radiographic findings suggested that crestal bone response around implants which were mobile versus immobile at placement was similar. CONCLUSIONS: Although implant stability at the time of placement is clearly desirable as seen in the superior 3-year survival of stable implants, it may not be an absolute prerequisite to osseointegration or to long-term survival. Several factors may influence the decision to remove or replace a mobile implant. HA-coating significantly improved the performance of both mobile and immobile implants at placement to 3 years post-placement (P < 0.001). PMID- 11885181 TI - Stability of the bone-implant complex. Results of longitudinal testing to 60 months with the Periotest device on endosseous dental implants. AB - BACKGROUND: Maintenance of the health and integrity of the bone-implant complex (osseointegration) has been shown to be essential for long term success of root form, endosseous dental implants. If reliable clinical indicators of adequacy of the bone-implant complex existed, they could stimulate new and innovative early intervention research to arrest of reverse early deterioration of the bone implant complex. In the absence of such indicators, this has been problematic. The Periotest may have the potential to provide this information by indirectly assessing the status of the bone-implant complex. However, little information is available that documents either the capability of the Periotest to reliably assess changes of the bone-implant complex or the "normal variations" in Periotest values (PTVs) for both HA-coated and non-coated implants. METHODS: The purpose of this paper was to document changes in PTVs as influenced by various implant surfaces, implant designs, and bone densities. The mean PTVs recorded for each visit, for all implant types and bone densities, were combined to provide an overall average PTV (A-PTV). The changes in stability (PTVs) were analyzed using a generalized linear model (GLM) with repeated measures (Hotelling's Trace). RESULTS: The A-PTV for all implants over all visits was -3.5. The mean PTVs ranged from -4.2 (SD = 2.4) at uncovering to -3.9 (SD = 2.9) at 60 months. All implants in bone qualities 1 and 2 (BQ-1 and BQ-2) became more stable over time, while those in bone quality 3 or 4 (BQ-3 and BQ-4) showed a slight decrease in stability. In BQ-1, the mean PTVs increased from -4.7 at uncovering to -4.9 at 60 months. A similar increase in stability occurred in BQ-2 (-4.1 at uncovering to 4.4 at 60 months). In BQ-3, the stability of the implants decreased over time ( 3.6 at uncovering to -2.9 at 60 months), with similar changes recorded for BQ-4 ( 2.5 at uncovering to -1.0 at 60 months). When comparing the stability of all HA coated with all non-coated implants, the HA implants became less stable (-4.4 to 3.4) over time, while non-coated implants showed an improvement in stability ( 3.5 to -4.5). The changes in stability found in BQ-1, BQ-2, and BQ-3 were similar, with HA implants becoming less stable and non-coated more stable. HA- and non-coated comparisons were not possible in BQ-4 since there were too few non coated implants placed in this type of bone. The HA-coated screw showed a decrease in stability when compared to the non-coated screw. CONCLUSIONS: Conclusions of the study are as follows: 1) PTVs are influenced by bone quality and surface coating of the implant; 2) the PTVs at the time of uncovering provide the best estimate of a clinically acceptable PTV for that bone-implant complex; 3) while the PTVs for any bone-implant complex may fluctuate +/- 1.0 around the uncovering PTV during routine healing and loading of the implant, a consistent shift toward a positive PTV that approaches "0" should be cause for concern that the bone-implant complex may be at risk of failure; 4) HA-coated implants became slightly less stable (more positive PTVs) over time, while the non-coated implants became more stable (more negative PTVs); and 5) until a "critical PTV" can be accurately identified, it is suggested that a consistent shift in recorded PTVs that exceeds the +3.0 value on the PTV scale should be viewed with concern for possible deterioration at the bone-implant complex. PMID- 11885182 TI - Periodontal-type measurements associated with hydroxyapatite-coated and non-HA coated implants: uncovering to 36 months. AB - BACKGROUND: While the use of hydroxyapatite (HA)-coated endosseous dental implants has gained in popularity over the past 10 years, the short-term and long term predictability and indications for their use remain highly controversial. Some reports suggest that the HA coating may separate from the substructure, undergo dissolution in tissue fluids, and/or contribute to rapid osseous breakdown around the implant. Other reports, however, relate favorable responses to HA-coated implants, which include rapid bone adaptation to the HA, greater stability at uncovering, and increased coronal bone growth. These contradictions may be related to differences in chemical composition of the HA on the implant surface. Most clinicians and researchers may agree that long-term, independent, scientific clinical studies are needed to compare HA-coated and non-HA-coated (titanium-alloy and CP-titanium) implants under the same conditions. Concerns appear in the literature that HA-coated implants experience greater breakdown because they are more susceptible to bacterial colonization due to their roughness and hydrophilicity. Some studies suggest that specific putative periodontal pathogens may adhere to the HA, thereby predisposing the implant to greater peri-implantitis than that experienced by non-HA implants. METHODS: A total of 32 clinical research centers, located in various geographic regions of the United States, were selected to participate in a comprehensive clinical study. More than 2,900 HA-coated and non-HA implants were randomized as to location within one of three jaw regions--maxillary anterior, mandibular anterior, and mandibular posterior--and followed for 36 months. It can be assumed that in each of these jaw regions, the conditions associated with both implant surface types would be similar enough to permit meaningful comparisons of periodontal-type measurements that have not previously been reported. Periodontal type measurements (gingiva, plaque, suppuration, and calculus indices; probing depth; attachment levels; recession; and keratinized tissue width) for each aspect of each implant (mesial, facial, distal, and lingual) were recorded at 3, 6, 9, 12, 18, 24, and 36 months following implant uncovering. The implant was considered the experimental unit for analysis using generalized estimating equation and repeated measure methods. Data for the four aspects of each implant, as well as measurements over time, were all clustered in the unit of analysis. RESULTS: The percentages of implants with zeros recorded for the indices was remarkably similar for both HA-coated and non-HA implants. While statistically significant differences were found for some of the measurements associated with HA-coated and non-HA implants under certain conditions, these differences were too small to be considered clinically significant. CONCLUSIONS: Overall, there was no clinically significant difference between the periodontal-type measurements for HA-coated and non-HA-coated implants followed for a period from 3 through 36 months. The concerns about HA-coated implants being associated with adverse periodontal responses for the HA chemical composition included in this study appear to be unfounded for a period of clinical performance up to 36 months. PMID- 11885183 TI - Clinical studies of endosseous dental implants: the good, the bad and the ugly. PMID- 11885184 TI - Variables affecting survival of single-tooth hydroxyapatite-coated implants in anterior maxillae at 3 years. AB - BACKGROUND: The development and expanded use of endosseous dental implants over the last two decades have been remarkably rapid. It is, therefore, imperative that the dental profession closely monitor the performance of root-form implants used in a variety of applications. The Dental Implant Clinical Research Group (DICRG) was established in 1990 by the Department of Veterans Affairs as a forum for conducting prospective, multidisciplinary, multicentered studies in the field of implant dentistry. The DICRG comprised 30 VA medical centers and 2 dental schools at the time of this study. This paper reports on the survival of hydroxyapatite (HA)-coated grooved implants used to replace single missing teeth in anterior maxillae at 3 years post-implant placement. METHODS: During a 4-year accrual period, a total of 247 single-tooth implant restorations were placed in anterior maxillae. This paper focuses on the survival of 222 implants (149 patients) for which 3-year data were recorded for the period from placement. Survival was examined with respect to patient demographics and health status, implant location, surgical variables, and 2-week post-placement use of chlorhexidine digluconate (0.12%) rinses. Implant stability was recorded using a hand-held probe. Periodontal-type measures were recorded and evaluated, and all complications related to osseointegration were noted. Failure was defined as removal of the implant for any reason. RESULTS: Establishment and maintenance of osseointegration at 3 years post-placement was 97.3%. During this 3 year period, 6 implants were removed due to either failure to osseointegrate or loss of osseointegration. Implant length correlated positively with 3-year survival (P = 0.003, exact test). The use of preoperative antibiotics was nearly significant to implant survival (P = 0.051. Pearson chi-square). Mean stability values (PTVs) increased incrementally from -4.5 at uncovering to +1.1 at 36 months, indicating a decrease in stability of the bone-implant-prosthesis complex. The most common complication was related to inadequate available bone to fully house implants. CONCLUSIONS: Three-year post-placement survival data suggest that the use of HA coated, grooved, endosseous implants to support maxillary anterior single-tooth replacements is a predictable and reliable procedure that can offer significant benefits. Longer implants demonstrated higher survival than shorter implants. The use of preoperative antibiotics was nearly significant to implant survival, and there was an increase in mean PTVs observed over the duration of the study. Further research is needed to assess stability of the hydroxyapatite-bone interface over time. PMID- 11885185 TI - The influence of smoking on 3-year clinical success of osseointegrated dental implants. AB - BACKGROUND: Health risks associated with smoking have been exhaustively documented and include increased incidence of periodontal disease, greater risk of osteitis following oral surgery, and compromised wound healing due to hypoxia. Information related directly to dental implants, although limited, points to higher rates of implant failures among smokers than non-smokers. This paper reports on long-term clinical outcomes of osseointegrated dental implants placed in smokers and non-smokers in a longitudinal clinical study of endosseous dental implants. METHODS: In 1990, the Dental Implant Clinical Research Group (DICRG) of the Department of Veterans Affairs (DVA) launched an 8-year, randomized, prospective clinical study of more than 2,900 endosseous dental implants in more than 800 patients at 32 study centers. Confounding variables, including smoking patterns, were recorded. For this report, new follow-up data were analyzed for two groups: 1) current smokers and 2) those who never smoked combined with those who quit. Most of the variables recorded for each implant were screened on a univariate basis as possible predictors associated with implant survival/failure. Those with P values less than 0.15 and those likely to be a factor of clinical importance were placed in a logistic regression equation and analyzed for a simultaneous effect on survival. A step-wise procedure was used to eliminate those variables that showed the least significance, until only those variables with a Wald chi-square of significance in the presence of others remained. The effects of clustering within patients and of unbalanced distribution within hospitals were standardized to facilitate analysis of influence of demographic variables. The GEE analysis was performed with the patient as the primary cluster. RESULTS: Current data do not support earlier findings that smoking contributes to early implant failure (placement to uncovering). A trend of greater failures in smokers appeared between the time after uncovering and before insertion of the prosthesis. Hydroxyapatite (HA)-coated implants had significantly lower failure rates. For the entire 3-year period, overall failures were significantly higher for smokers than non-smokers. CONCLUSIONS: Results suggest that increased implant failures in smokers are not the result of poor healing or osseointegration, but of exposure of peri-implant tissues to tobacco smoke. Data also suggest that detrimental effects may be reduced by: 1) cessation of smoking; 2) the use of preoperative antibiotics; and 3) the use of HA-coated implants. PMID- 11885187 TI - Responding to anthrax. PMID- 11885186 TI - Influence of two different approaches to reporting implant survival outcomes for five different prosthodontic applications. AB - BACKGROUND: Over the years, the definition of implant failure has varied, with some investigators accounting for all implants placed, while others discard failures that occurred before clinical loading. The influence of stresses transmitted to various bone densities, by different prosthetic appliances as well as the method used to determine failures, needs to be clearly understood. This paper reports on the influence of two different methods of determining 36-month survival of implants used to support different dental prostheses. METHODS: More than 2,900 implants with six different designs were placed in 829 patients at 32 study centers and followed for 3 years. The first method of determining survival accounted for all failures from placement through 36 months, while the second method counted only failures from post-loading of the prosthesis to 36 months. Survival curves were used to determine differences in survival outcomes for the two methods. RESULTS: For the maxillary single-tooth prosthetic application, implant survival from placement to 36 months was 94.7% when all failures were counted and increased to 98.3% with the post-loading method. For upper completely edentulous applications, implant survival was 85.3% with all failures counted and 95.6% with the post-loading method. This 10.3% difference is clinically important. The survival for implants in lower completely edentulous applications increased by 4.4% simply by using the post-loading approach. Implants used for upper posterior, partially edentulous applications involved only hydroxyapatite (HA)-coated implants, and the survival rates were similar (96.4% for all implants and 98.2% for the post-loading method). The difference in reported survival rates for implants in the lower posterior, partially edentulous application was 5.8%. Since failures tended to occur in the earlier phases of treatment, the post loading approach always resulted in more favorable survival data. With the post loading approach, valuable information related to implant performance before loading is lost. In all comparisons, HA-coated implant survival was always better than non-HA implants. Clinical investigators should clearly state the method used for determining failures. For all implants included in the study, survival curves illustrated different failure patterns for each method of determining overall survival. CONCLUSION: Reporting of implant survival rates based on the post loading method provides more favorable survival rates; however, accounting for all implants provides a more accurate method of determining survival. PMID- 11885188 TI - The TEREC connection. PMID- 11885189 TI - Designing your lab team. PMID- 11885190 TI - College quality. The same old way (not!). PMID- 11885191 TI - Vo-tech prepares the pros. PMID- 11885192 TI - The art and science of diagnostic waxing. PMID- 11885193 TI - The interdependence of dentist and laboratory in successful management of a rehabilitation case. Part 2. PMID- 11885194 TI - Life at the bench. Fired. PMID- 11885196 TI - Diagnostic orthotics to establish the functional mandibular-maxillary relationship for orthodontic corrections. AB - Under optimal anatomic and physiologic circumstances, there exists a harmonious and functional balance between the occlusion, muscles of mastication and joint relationships referred to as a functional bite relationship. A diagnosis and treatment to the habitual bite relationship is provided if the function of the joints, muscles and associated structures is determined to be normal. If compromises of form or function are determined, a diagnostic orthotic is an essential tool to evaluate why the habitual bite position is not the best functional bite position and to also help determine a better functional relationship to avoid tissue and system compromise. The benefit of the diagnostic orthotic is the ability to establish an arbitrary reversible diagnostic relationship, evaluate the tissue and system response to this new association over time, and determine if the new treatment position is better for the patient than the habitual bite association. PMID- 11885195 TI - The product of partnership. PMID- 11885197 TI - Treating to beautiful faces, healthy TM joints, faster treatments and stable arches. PMID- 11885199 TI - The interdependence of dentist and laboratory in successful management of a rehabilitation case. Part 1. PMID- 11885198 TI - Do your doctors think you're worth it? PMID- 11885200 TI - Software essentials for lab success. AB - "Look around before you make a decision," Whelan said. "Take the time for a hands on trial of different systems. Ask other labs using the system what they think of it. After all, this is an important decision for the long-term health of your business and one you will live with for a long time." PMID- 11885201 TI - A framework for success. Part 2: The joy of (understanding) money. PMID- 11885202 TI - Training in the dental laboratory: the presentation. PMID- 11885203 TI - Six attitudes that destroy teamwork. PMID- 11885204 TI - What's driving the industry. PMID- 11885205 TI - Understanding dental system economics. PMID- 11885206 TI - Education. Committed to change. PMID- 11885207 TI - Lab on the water: Town & Country Dental Studios. PMID- 11885208 TI - Making the most of the Economic Growth and Tax Relief Reconciliation Act of 2001 (EGTRRA). PMID- 11885209 TI - Creating lab opportunity with products, sales support and communication. PMID- 11885210 TI - Continuum of care for Alzheimer's disease: a nurse education and counseling program. AB - This study evaluated a one year long course education and counseling program with 93 family caregivers of elders afflicted with Alzheimer's disease. The elders had received treatment for agitation in an inpatient setting and were subsequently discharged to the caregivers' home. Caregivers were randomly assigned to an experimental group (n = 68) and a control group (n = 25). Baseline assessments (Time 0) were conducted while the elder was an inpatient. Postdischarge interventions and assessments were conducted at 2 weeks, 6 weeks, 12 weeks, and 6 and 12 months (Times 1-5 respectively). There were no significant treatment effects for care recipient agitation, caregiver stress, depression, and physical health, and no significant differences between groups in rates of institutionalization for afflicted elders. Longitudinal data, however, revealed several important trends. Afflicted elders' agitation rose steadily for control group subjects at Times 3 through 5 but declined for experimental group subjects. Caregiver depression increased for control group subjects at Time 5, but declined for experimental group subjects. Caregiver physical health declined for control group subjects at Times 4 and 5 but was maintained for experimental group subjects. A significantly higher number of afflicted elders were still at home among experimental group subjects at the end of the one year study. The difficulties in demonstrating efficacy of interventions with family caregivers of Alzheimer's disease are discussed. Finally, the issue of data collection being perceived as support by control group subjects is evaluated. PMID- 11885211 TI - Relationships of religion, health status, and socioeconomic status to the quality of life of individuals who are HIV positive. AB - The present study tested three hypotheses about the quality of life of individuals who are HIV positive. It was hypothesized that quality of life among HIV-positive individuals would be directly related to their (1) health status, (2) religious affiliation, and (3) religious faith. A correlational design was used with a nonrandom sample of 40 subjects (32 males and 8 females) who were HIV positive. Bivariate analyses were conducted to obtain intercorrelational among several independent variables, including two measures of religion (religious affiliation and a composite measure of religious faith), number of symptoms, level of physical functioning, and various demographic measures, including socioeconomic status. Stepwise regression confirmed all three hypotheses, revealing that four independent variables made significant, positive contributions to subjects' scores on the Quality of Life Index (QLI). These were socioeconomic status, religious affiliation (affiliation vs. no affiliation), religious faith, and a combined measure of health status based upon the participants' number of symptoms and Karnofsky Performance Status. The other independent variables (age, ethnicity, and gender) did not make significant contributions to the regression model, accounting for only 2.3% of the variance in the QLI. PMID- 11885212 TI - Is codependency a meaningful concept? AB - The concept of condependency has achieved a prominent place in the psychiatric, psychological, and addiction literature in a remarkably short period of time. Although the term was first developed in the substance abuse treatment arena, specifically referring to the wives of men who abuse alcohol, codependency has more recently been used almost generically to describe a dysfunctional style of relating to others (Irwin, 1995). The manner in which definitions of codependency have become increasingly inclusive are probably related to continuing input from the both the fashionable self-help movement and from some psychiatric perspectives. The purpose of this article is to review proposed definitions of codependency, discuss issues related to the validity of the codependency construct, and summarize efforts aimed at producing instruments to measure codependency. Additionally, I will address implications of this concept as related to psychiatric nursing education, practice, and research. PMID- 11885214 TI - Problems of self-regulation: a new way to view deficits in children born prematurely. AB - While survival rates for the smallest infants are increasing, so is the rate of disability. Low birth rate children are at increased risk for psychiatric and behavioral symptoms especially those related to attention deficit and hyperactivity disorders. Researchers have demonstrated that even "normal" low birth weight (LBW) children receive special educational services at an alarming rate. Little is understood about the processes responsible for these academic delays. Preventative interventions cannot be implemented without understanding the underlying developmental processes. The study of self-regulation (SR) of cognition and the factors that may influence the development of regulatory capacity are suggested as a way to frame future work. SR of cognition refers to one's ability to select and use information appropriately. Problems in the development of the self-regulation of attention may explain deficits in the acquisition of cognitive skills as well as other deficits. The argument is made that specific variables may directly, indirectly or in both ways influence mechanisms and processes underlying the development of attention in LBW children. It is proposed that studying the SR of cognition provides a potentially useful and powerful focus for intervention research. PMID- 11885213 TI - Factors affecting substance abuse treatment completion for women. AB - Substance abuse by women is considered an individual pathology, and the larger social processes of recovery are seldom explored. This research study examined social factors that influenced completion of an outpatient women-centered substance abuse treatment program. The treatment records of a group of 15 women who completed the program were compared with a group who did not complete the program. More completers had previous life successes in the areas of education, job skills, and employment history. Completers also had fewer children, less involvement with child protective services, and lower levels of chaos, a construct that included the presence of two of any of the following in women's lives: child protective services, homelessness, psychiatric diagnosis, or domestic violence. Completion of substance abuse treatment seems more likely for women with personal and social resources. If programs are to be successful, adequate funding must be provided for both assessment and support of the social problems encountered by the most vulnerable women. PMID- 11885215 TI - Patient education about schizophrenia: initial expectations and later satisfaction. AB - This study investigated patients' expectations prior to participation in an education program about coping with schizophrenia, and their evaluations of the program upon its completion. Adult inpatients diagnosed with schizophrenic disorders (N = 123) responded anonymously to a preintervention expectation measured and a postintervention evaluation questionnaire. Results point to high expectations of this illness self-management education program, and a high level of satisfaction upon its completion, with a self-fulfilling prophecy effect, in which those with high expectations later reported greater satisfaction. Patients perceived, however, a differential level of helpfulness of the program's nine content areas, and rated learning about diagnosis and medication management as most helpful. Content areas that were rated less helpful included prevalence of schizophrenia, its psychosocial rehabilitation, and use of community resources. Implications for clinical practice in patient education are identified and discussed. PMID- 11885216 TI - About this issue on youthful offenders. PMID- 11885217 TI - An overview of forensic psychiatric care of the adolescent. AB - The field of forensic psychiatric nursing is a relatively recent addition to the specialty of psychiatric nursing. This paper provides an overview of forensic psychiatric care of adolescent patients. It describes the juvenile justice system, identifies various theoretical models useful for understanding youthful offenders, and explores risk and protective factors. Psychiatric comorbidities and treatment considerations also are presented. PMID- 11885218 TI - Healing the victim, the young offender, and the community via restorative justice: an international perspective. AB - The 1990s saw the enactment of much "get tough with young offenders" legislation in the United States. At the same, problems with our present punishment and treatment model, in which many youngsters cycle repeatedly through the justice and mental health systems, raised interest in restorative justice, a community based alternative model emphasizing a balanced, negotiated approach to the needs of victims, offenders, and the community. After summarizing the philosophical bases underlying both models, this article describes the practice of restorative justice in New Zealand, where it was pioneered. Restorative justice has special relevance for Maori community in New Zealand and minority communities in the United States, where youth are consistently overrepresented in the courts, detention centers, and jails, and in which the juvenile justice system is seen as hostile and biased. Outcome data from New Zealand and early outcome research from the United States suggest that the restorative model, in which offenses are understood as a breakdown in social bonds, offers a hopeful alternative for offending youngsters, their families, and their communities. PMID- 11885219 TI - Adolescent addiction and delinquency in the family system. AB - Drug abuse and delinquent behavior in adolescents have been closely linked in the literature. It is possible that these links are not causal, but rather covariates of the common factor of family. This paper seeks to examine the behaviors of addiction and delinquency within the framework of family systems theory. PMID- 11885220 TI - AIDS and drug use prevention intervention for confined youthful offenders. AB - This paper presents pilot work to develop and test an intervention designed to reduce high-risk behaviors that lead to the development of HIV infection in adolescents. The intervention was designed for youth that have high-risk behaviors, a history of drug us or both. The intervention was based upon Peer Counseling and Leadership Techniques (PCLT) that have been successfully implemented with adult offender populations. These methods were modified to address the specific aims of the pilot study were two fold: (1) to evaluate the modifications of the intervention, and (2) to test the short-term outcomes regarding knowledge acquisition, enhancement of self-esteem, and readiness to change behavior. PMID- 11885221 TI - The correctional Officer's role in Mental Health treatment of youthful offenders. AB - This paper explores issues specific to the provision of effective mental health treatment for juvenile offenders within juvenile justice departments. A multifaceted treatment approach in this setting should include security officers as well as the customary mental health and medical professionals. Confidentiality issues direct correctional policies that require medical, mental health and security records to remain separate. Such limitations in the exchange of information reduce the likelihood that correctional officers would be considered as part of the treatment team. With high inmate to professional staff ratio and the growing attention to outcomes, this approach needs to be considered. PMID- 11885222 TI - Using creative arts to build coping skills to reduce domestic violence in the lives of female juvenile offenders. AB - The development of gender-specific programming is gaining attention as the approach that most effectively addresses the needs of female youthful offenders. This study provides a profile of female juvenile offenders, their problems and needs, and suggests psychoeducational approaches for building coping skills to reduce cycles of domestic violence. This is particularly important within a system designed primarily for their male counterparts. PMID- 11885223 TI - The role of contextual, child and parent factors in predicting criminal outcomes in adolescence. AB - This study examined predictor variables that would contribute toward an explanatory model linking child maltreatment and the outcome of crime seriousness. A secondary data analysis was completed on a random sample of youth committed and detained by a juvenile justice system in the northeast United States. Although existing data indicate a predisposition to crime and violence among youth that are maltreated, more research is needed to determine the exact nature of the link, as well as the need to determine the relationship of associated factors. The aim of this analysis was to identify key contextual, child, and parental factors related to maladaptive behavior in abused and neglected adolescent offenders. Logistic regression was used to predict serious criminal outcomes. Overall, 73% of the cases were correctly classified. Ten variables remained in the model to predict serious juvenile criminal behavior. Youth who had been exposed to community violence or who had a mother with mental illness were four times as likely to commit serious criminal behaviors. PMID- 11885224 TI - The challenge of juvenile justice: advocating for troubled children in trouble. AB - Psychiatric nurses have long advocated for children with mental disorders, but few have gone behind bars to advocate for these youth. This paper offers suggestions for advocating on behalf of the most underserved of children-youthful offenders. Efforts in Maryland, as an example, portray conflicting convictions: to punish or to treat. These conflicting convictions permeate our juvenile laws, juvenile facilities, and treatment programs. A discussion of successful programming demonstrates the strengths of advocacy and the difference one person can make. Steps are outlined to assist nurses in the development of activist roles in advocating for these troubled youth. PMID- 11885225 TI - Effect of self-brushing with acidulated phosphate fluoride (pH 5.6) on dental caries in children. AB - The aim of this work was to study the effect of a preventive program on the dental plaque and on the caries incidence in school children. The program comprised weekly supervised self brushing with acidulated phosphate fluoride (APF) gel (pH 5.6; concentration: 4520 ppm of ion F-). The program involved 240 children of 1st., 3rd and 5th grade of a primary school in the City of Buenos Aires (Argentina). The population was divided in 2 groups: A (experimental) and B (control). Ninety children from both groups (45 experimental and 45 control) were submitted to baseline clinical examination (DMFT and plaque index) and microbiological analysis (total streptococci, St mutans and St. mutans and St. mutans percentage). Group A was then submitted to a preventive program which included self brushing with APF gel (4520 ppm of ion F-). The 90-children sample was monitored after 1 and 2 years of program. Results were statistically processed and they revealed the following: a--DMFT was significantly greater in the control group than in the experimental group after 1 and 2 years of program; b--an 81.43% reduction in caries increment rate at the end of the 2-years program in the experimental group as compared to the control group; c--a rise in the number of colonies of total streptococci and of St. Mutans; d--a reduction in the % of St. mutans in the total streptococci flora in the plaque of children in the experimental group; e--the presence of St. mutans colonies featuring a rough surface; f--the effectiveness of the program in the modifying the profile of the diagnosed dental pathology. PMID- 11885226 TI - Effect of chronic constant light on sensitivity of rat submandibular gland. AB - A consistent difference in the secretory response between submandibular (SM) glands of rats maintained under constant light (CL) during 50 days and those of rats under a photoperiod (14 h light: 10 h dark) was found. We have used alpha 1 adrenergic, muscarinic, peptidergic and beta-adrenergic secretagogue agents to study the secretory response of rat SM glands "in vivo". The response to phenylephrine, methacholine and substance P, was increased by exposure CL, while that to isoproterenol was diminished. The changes in the sensitivity of the secretory response from SM gland of rats under CL might be related to changes in the normal interplay of various receptors as well as to possible alteration in the intracellular signal transduction. It may represent and adaptive process of the nervous control of saliva secretion by environmental light and be of physiological and clinical interest. PMID- 11885227 TI - Bone growth is impaired by uranium intoxication. AB - Acute and chronic uranium intoxication leads to the inhibition of bone formation and impaired bone modeling and remodeling. As these are processes directly involved in bone growth the aim of this paper is to present a biometric study of bone growth--tibiae and mandibles of rats intoxicated with uranium. Wistar ratios weighing 60-80 g were used as follows, a) one intraperitoneal injection (IPI, 2 mg/Kg of body weight)) of uranyl nitrate; b) 30 daily applications on the dorsal skin of aliquots of a mixture of U308, concentrated at 2% and at 4%--percutaneous absorption(PA)-. Tibia and mandible length were smaller in both experimental groups than in their respective controls. Some of the mandibular parameters were lower in intoxicated animals than their controls which in turn results in the alteration of the mandibular shape. We conclude that impairment in bone growth can be achieved by uranium intoxication. PMID- 11885229 TI - No association between secretor status of ABO blood group antigens and juvenile periodontitis. AB - The aim of this study was to determine the association between secretor status of ABO blood group antigens and localized juvenile periodontitis (LJP). Forty-three patients with LJP (mean age 20.8 years) diagnosed according to Baer's criteria were selected. Thirty two periodontally healthy normal subjects (mean age 24.7 years) were use as control. Samples of blood and saliva were collected from patients and from control subjects. ABO blood group was determined by agglutination of erythrocytes with appropriate antisera. Determination of soluble ABO antigens in saliva was made by the haemagglutination inhibition assay. Subjects with O blood group were most frequent in both groups. The distribution of blood groups, and secretor and non-secretor status of ABO group antigens in LJP patients and control subjects was not significant. The results provide support for the hypothesis that there is no association between non-secretor status and LJP. PMID- 11885228 TI - Antimicrobial power of composite resin adhesion systems. AB - The adherence of microorganisms to dentin that had been contaminated and then treated with conditioning substances was evaluated. The germicide effect of those products and their possible substantivity was also evaluated. Dentin slices were contaminated with Candida albicans, Streptococcus mutans and Actinomyces naeslundii and then treated with the following substances: 1. Experimental Blue Solution A, 2. Experimental Red Solution B; 3 Tubulicid Blue Label; 4. Tubulicid Red Label; 5 Scotchprep Dentin Primer; 6 ventura Dentin Bond Cleaner; 7 ventura Dentin Bond Primer, 8. Gluma dentin Bond; 9 Tenure Conditioner; 10. All Bond Dentin Conditioner; 11 Syntac Primer; 12. Clearfil New Bond acid + adhesive; 13 Prisma Universal Bond 3 Primer; 14. Denthesive Cleaner; 15. Control (Distilled water). Adherence was evaluated using scanning electron microscopy and viability tests were performed. Substances 1, 2, 3, 4, 5, 7, 8, and 11 impair adherence; the control and substances 6 and 9 produced no afforded intermediate results effect while substances 10, 12, 13 and 14. Viability tests afforded results similar to those obtained for adherence. Several substances that are used for dentin treatment or for dentin priming for adhesion could be useful in preventing microorganism viability and so contribute to the protection of pulp vitality. PMID- 11885230 TI - Use of Zircalloy to induce bone regeneration. A 2-year follow-up study. AB - In a previous study we analyzed the behaviour of Zircalloy 4 particles employed in patients with significant loss of periodontal bone support which threatened the permanence in situ of the tooth. We herein present the results obtained after regular examination over a 2-year period of 6 patients submitted to this treatment. Our results reveal that all patients exhibited excellent gingivo periodontal health with an increase in clinical insertion of 7 mm +/- 1 and a marked reduction in motility from 3 to 1. None of the cases featured added inflammation or expulsion of the implant material. In one of the cases it was possible to study the histology of the area by punch biopsy with a disposable needle (Sherwood-Medical). Analysis of the sample revealed the presence of bone tissue in intimate contact with implanted metallic particles of Zircalloy 4, producing osseointegration. This osseointegration would lead to filling of the periodontal bone defect. Macrophages loaded with metallic particles were found in the vicinity of particles which were not osseointegrated. These features may correspond to superficial areas related with soft tissue as observed in our study on alveolar filling in rats with the same type of particles. PMID- 11885231 TI - Depressed eruption dental rate in rats with hemodynamically-mediated acute renal failure. AB - The effects of acute renal failure on the impeded (IER) and unimpeded (UER) eruption dental rate and attrition rate (AR) were investigated. Adult female Wistar rats were injected with 125 mg/kg b.w of human methemoglobin (M-Hb) in order to induce a first episode of hemodynamically-mediated acute renal failure (H-ARF). Ten days after the injection of M-Hb, other groups of rats received another equal dose of the drug in order to induce a second episode of H-ARF. A group of six animals was pair-fed daily and individually with rats of M-Hb groups. Evaluation of renal function, histopathology studies, IER, UER, food intake (FI), AR and body weight gains was performed at different times after the first and second injections, of M-Hb. Treatment induced transient increases in plasma urea concentration and urine volume, and significant depression in urine osmolality, body weight gains, IER, UER and AR. In every case, the maximal effect of the first injection of M-Hb on the individual parameters was always greater than that of the second injection. Histologic sections showed interstitial cellular infiltration, desquamation of the proximal tubular epithelium and collapse or dilation of the tubular lumen. The functional values of kidney, histologic findings, IER, UER and AR of the pair-fed rats were not significantly different from control values. The results of the present study indicate that dental eruption rate (IER and UER) is relatively low in uremic rats with kidney tubule lesions and that both parameters are related. PMID- 11885232 TI - Additive effects of dietary protein and energy deficiencies on mandibular growth in the weanling rat. AB - Dietary protein restriction adversely affects mandibular growth in the weanling rat. Protein deficiency is usually accompanied by reduced food intake which, in turn, induces energy deficiency. The present study was thus designed to dissociate the effects of dietary protein and energy deficiencies on the growth of the mandible in rapidly growing rats. Four groups of Sprague-Dawley rats aged 30 days were fed a normal diet, a low-energy diet, a low protein diet, and a low protein and low-energy diet for 20 days. Rats were sacrificed at the end of experimental period and body weight and mandibular dimensions were recorded to evaluate body growth and mandibular growth. The growth of the mandible was affected almost in the same order of magnitude by both protein and energy restrictions. When both were applied together, mandibular growth was even more severely affected. Two way analysis of variance revealed the absence of synergism between variables, indicating that the negative effects of dietary protein and energy restrictions on mandibular growth could be considered to be additive. PMID- 11885233 TI - Study on gutta-percha points using scanning electron microscopy and analysis with electron microprobe. AB - A comparative study of the apical morphology and contour was performed with the scanning electron microscope at 45x, 70x and 300x, while the chemical composition of eight gutta-percha point brands was surveyed with an EDAX 9100 electron microprobe. They were classified according to their apical morphology as conical (25%) and truncate (75%), based on their contour as regular (50%) and irregular (50%) and according to their surface as without defects (50%) and with defects (50%). The analysis of inorganic compounds revealed the presence of varying proportions of Zn, Ba, Si, Mg. Ca, P, Cl and Al, possibly implying that the presence of Ba and Al could interfere with postendodontic repair or at least irritate the periapical area if inadvertently overfilled. PMID- 11885234 TI - Extraction and quantification of heparin in "clinically normal" and inflamed gingiva of the dog. AB - Heparin present in tissue has been reported to be a potential locally active agent responsible for bone resorption by the stimulation of collagenase via osteoclast activation and increased collagenase synthesis by the osteoblast. The purpose of this study was to extract, identify and quantify heparin in the "clinically normal", mildly and severely inflamed gingiva of Beagle dogs, using a simple and reliable technique. The extraction was carried out by homogenization, fat elimination, proteolytic digestion and precipitation, with organic solvents. Identification was performed by microelectrophoresis conducted on an agarose coated microscope slide. Quantification was performed by measuring the optical density of the metachromatic toluidine blue stained spot an comparing with the standard reference curve of heparin run simultaneously. The results showed that the difference in concentration of heparin in units per gram of wet tissue, was not significant when the "clinically normal" (26 +/- 1.9) was compared with mildly inflamed (24.4 +/- 4.7) gingiva. However, the concentration of heparin in severe gingivitis (79 +/- 7) was significantly higher. Gingival heparin could play important role in the established periodontal lesions, acting as a local factor or co-factor in periodontal bone destruction. PMID- 11885235 TI - A method for the quality control of osseointegration in endosseous implants. AB - Metallic dental and orthopedic implants are essential therapeutic tool. The biologic success of an implant involves intimate contact between the implant and vital bone tissue, an event which has been termed osseointegration. The aim of the present study was to study the biocompatibility and biomechanic properties of different implant materials. Zircalloy 4 metallic cones (1.8 mm base x 2 mm height) were implanted in the diaphysis of the tibiae of 6 Wistar rats (90 g body weight) under i.p. Ethyl Urethane anesthesia. The animals were killed 30 days postimplantation, the tibiae were resected, and radiographed. One side chosen at random was used for the biomechanical study; whereas the other implant of each animal was processed for embedding in methyl-methacrylate. The bond of the implanted material to bone tissue was monitored immediately after resecting the tibiae by applying an extraction force to the base of the cone via a device designed ad hoc connected to a testing machine. Histological and radiographic analyses revealed the presence of bone tissue in contact with the implant surface. A force of about 35 g was necessary to separate the implant from the bone tissue. The system proposed renders possible the study of biocompatibility in histologic terms and in terms capacity to bond to bone tissue and could be a valuable research tool and the basis for quality control of all types of material, metallic or otherwise, used in endosseous implants. PMID- 11885236 TI - Sickle cell anemia oral manifestations in a Venezuelan population. AB - Sickle cell anemia (SCA) is a well known hemoglobinopathy which results from a substitution of amino acids in the polypeptidic chain. SCA was considered endemic in certain areas of the world. It has been recognized now that it may have a wide geographic distribution. Few studies have dealt with dental manifestations or complications of SCA (Cox and Soni, 1984). Nevertheless none of them have showed epidemiological data for a large series of oral manifestations. To date, no epidemiological data of our country is available in the literature. The aim of this study was to determine the oral manifestations of SCA in a Venezuelan population. Seventeen patients affected were examined at the University Hospital and the Dental Clinic. Age ranged between 1 1/2-48 years. Each patient was haematologically diagnosed by hemoglobin electrophoresis and only homozygous individuals were selected. Each patient was analyzed according to general clinical history, as well as, dental history; clinical and radiological examination using periapical, panorex and bite-wings radiographs. Our results showed that the most affected group was between 20 to 30 years (41.18%). According to sex, females were more affected than males (64.71%). The most common phenotype was mestizo (47.31%). The most frequent type of hemoglobinopathy was Hg SS and Hg SS-F. The most common soft tissue oral manifestation was buccal mucosa pallor in 77.05%. In addition, the hard tissue findings involved enlarged medullary spaces (70.58%). Cicatritial infarcts were present in 77.05% of cases and the step-ladder effect was demonstrated in 82.35% of cases. Our observations could be due to genetic, environmental, nutritional and geographical factors. PMID- 11885237 TI - Effect of compressive forces on a bone modelling surface. AB - The work presented herein, is an experimental study on the effect of an orthodontic appliance with a helicoidal spring designed to exert force toward palatine--i.e. in the opposite direction to the natural vestibular drift--on a bone remodelling surface. The appliance consists of two stainless steel molar bands, with a horizontal bracket tube welded to their palatal aspect through which the arms of the helicoidal spring are passed. Wistar rats, 250 g body weight, were fitted with the device for 48 and 96 hours. One group of rats was administered two doses of tetracycline hydrochloride prior to device placement, in order to label mineralizing fronts. Histomorphometric studies of the periodontal wall of the palatine alveolar bone showed a marked increase of bone resorption at both experimental time points together with an increase in the number of osteoclasts, and no tetracycline labelling after 48 hours. The results show that compressive forces are capable of stimulating resorption, even on bone modelling surfaces. The pressure applied would stimulate osteoblasts to send out signals for osteoclast recruitment and activity. PMID- 11885238 TI - Longitudinal study of periodontal condition in students of the Dental School the University of Buenos Aires Argentina. AB - The aim of the present study was to describe, clinically and radiographically, the gingivoperiodontal condition of students (young adults) entering the Dental School and reassess it 2 years later. Four hundred and seventy-five students, 147 male and 328 female (mean: 19.66 years) were studied (Initial examination: IE); two years later, 240 students 76 male and 164 female were reevaluated (Final examination: FE). The clinical examination include Plaque Index (P1I), Gingival Index (GI), Bleeding on Probing (BOP), Probing Pocket Depth (PPD), and Attachment Level (AL). Alveolar bone level was measured on bite wing radiographs. At the IE, 46.5% of the sites had no visual signs of inflammation and 65.3% did not bleed on probing, however 91% of the students had at least one site with BOP. (Prevalence). Of the total sites probed 99.47% had crevices between 1 to 4 mm. 92% had AL between 0 to 1 mm. And only 7.9% had AL > 1 mm. 49.6% of the students had at least one site with AL > 1 mm. Prevalence of alveolar bones loss > 2 mm. was 2.95% and in no case measurements exceeded 3 mm. Distal of teeth 16 and 36 showed the highest P1I, GI, PPD and AL, therefore these teeth showed the highest risk of pathology in the population studied. In the final examination 70.5% of the sites had GI 0 and 73% did not bleed on probing. 90.4% of the students had at least one site with BOP. No statistically significant difference in PPD nor in AL was found between IE and FE. In this population of young adults prevalence of gingivitis was high but severity was low. Prevalence of loss of attachment was near 50% but only 7.9% of the sites had AL > 1 mm. After studying dentistry for two years the prevalence of gingivitis was similar but the severity was lower. Periodontal condition was not substantially modified. PMID- 11885239 TI - Evaluation of serum and saliva components in candidosis patients. AB - The serum and saliva components of 36 chronic Candidosis patients, both male and female, ages 38-82 who attended the Department of Clinical Stomatology were studied. Total Mucous Lesion Index (TMLI) and salivary flow rate were assessed. The following parameters were evaluated: iron bound protein, unsaturated iron binding capacity, peroxidase activity, protein content, OSCN-, SCN-, IgAs, Candida and St. mutans levels and lactobacilli activity. Candidosis patients exhibited higher Candida CFU values and increased activity of the peroxidase system (p < 0.05) whereas unsaturated iron binding capacity was significantly lower as compared to healthy subjects (p < 0.05). Furthermore, TMLI and Candida CFU values were higher in diseased subjects wearing complete prosthesis as compared to those without complete prosthesis. Diseased subjects with < 1.2 ml/min salivary flow rate exhibited even greater differences with control. This subgroup exhibited a marked reduction in IgAs. The serum components assayed were iron bound protein, unsaturated iron binding capacity, IgG, IgA and IgM. Unsaturated iron binding capacity was significantly lower in the Candidosis group (p = 0.03). Subjects suffering from oral Candidosis display deficiencies in some of their saliva components, evidencing impaired oral defense capacity. PMID- 11885240 TI - Age-related histochemical and immunohistochemical changes in human labial salivary glands. AB - We here in determined the histochemical and immunological changes that salivary glands experience as a result of the ageing process. Samples from the inferior lip of children, youngsters and young adults were analyzed by histochemical techniques, Periodic-acid-Schiff, (PAS) Alcian blue (AB) (pHs 1.0 and 2.5) and Toluidine blue for mucosubstances and immunohistochemical staining of S 100 protein and cytokeratin 20, avidin-biotin system (DAB). In children, the techniques used evidenced various reaction degrees in the acinar cells, even within a single acinous. They displayed a slight metachromasia and were alcianophilic at pH 2.5. In youngsters, and especially in adults, glands showed a notable PAS positivity and alcianophilia at both pH levels, and an intense metachromasia. PS 100 was positive in the basal area of the acini and serous demilunes of all groups, the reactivity being higher in adults. Cytokeratin 20 was better observed in ductal cells from children glands. These findings suggest modifications at cytological level and in the chemical composition of the secretory granules, indicating possible functional variations in lip salivary glands related to the ageing process. PMID- 11885241 TI - Structural and cytochemical modifications in the lingual glands of the newborn chicken irradiated with He-Ne laser. AB - Despite the increasing and successful use of laser in Medicine and Odontology, the possible iatrogenic and otherwise deleterious side effects of this radiation remain mostly unknown. In previous studies, it was shown that both the embryonic and the post-hatched chicken constitute reliable experimental models for this type of studies. Hence, the purpose of the present work was to analyze the structural and cytochemical alterations of the lingual glands of the newborn chicken irradiated with low energy He-Ne laser. This laser produced regressive structural changes of the glands towards the embryonic stage as well as hyperplasia of the reserve glandular basal cells. Furthermore, a decrease in the glycoprotein content and a rise in the sulphated glycosaminoglycans were also found. These results corroborate the pathogenic effects of the He-Ne- laser on the experimental model employed and, at the same time, emphasize the importance of considering, regarding clinical applications, possible previous neoplastic alterations as well as adverse reactions which might appear once laser therapy has been installed. PMID- 11885242 TI - A comparative study of oral lichen planus and leukoplakia in two Argentine populations. AB - Oral Lichen Planus and Leukoplakia are two precancerous lesions of great relevance in oral pathology. A total of 4183 patients from the National University of Cordoba (UNC) and 4838 patients from the University of Buenos Aires (UBA) who had been admitted to the corresponding Oral Pathology Departments were analyzed. Of the total number of patients, 476 corresponded to Lichen Planus cases and 418 to Leukoplakia cases. Of the 476 Lichen Planus cases, 330 came from UBA and 146 from UNC, whereas of the 418 cases of Leukoplakia, 284 came from UNC and 134 from UBA. These differences were statistically significant (p < 0.02). Distribution according to sex and age was similar for Lichen Planus and Leukoplakia patients from both Oral Pathology Departments. The association between diabetes and Lichen Planus was similar for both centers, 11.5% for UNC and 14% for UBA. Similarly, no differences were found in terms of the association with tobacco consumption and dental microtrauma. Twenty-two percent of UNC patients were smokers whereas only 11% of UBA patients were smokers. This finding could explain the larger amount of Leukoplakia in UNC. The differences in the incidence of Lichen Planus could be attributed to the fact that the Buenos Aires population is under greater stress and the higher incidence of Leukoplakia in UNC could be related to the smoking habits of this population. PMID- 11885243 TI - Simultaneous PAGE, immunoblotting, and immunohistochemical analysis of differentiation associated keratins in lesions of the oral mucosa. AB - The expression of differentiation associated high PM Keratin polypeptides of the oral mucosa lesions were studied by immunohistochemical and immunoblotting techniques applied to adjacent sections of each biopsy specimen. The material studied included specimens of leukoplakia, verrucous carcinoma, squamous cell carcinoma, adenocarcinoma and keratoacanthoma. Little or no expression of 65-67 Kd keratins was evident in squamous cell carcinoma and adenocarcinoma. Hyperkeratotic (both benign and dysplastic) lesions such as verrucous carcinoma, leukoplakia, and keratoacanthoma, showed great variations in the intensity of 65 67 bands and a very irregular immunohistochemical staining pattern. Increased amounts of horny substance was usually accompanied by absence of, or decreased expression of 65-67 Kd keratins, thus indicating a change in the polypeptide composition of the horny layer in pathological conditions of the oral epithelium. PMID- 11885244 TI - Embryogeny of human labial glands: a structural, ultrastructural and cytochemical study. AB - A histochemical study of labial glands was performed to compare the different stages of differentiation with those of lingual glands previously studied. Labial glands of 8 to 32 week old human fetuses were analyzed with Hematoxylin/eosine, PAS, Cason, Alcian blue, Toluidine blue, methenamine/silver, TEM and Ruthenium red techniques. At 8-10 weeks various differentiation phases of cell cords originated in the epithelium of the labial mucosa were observed. Acinar buds had PAS positive, alcianophilic and metachromatic material in the lumen of 14 week labial glands. The excretory ducts featured similar characteristics. At 24 weeks groups of mucous and seromucous acini were identified and the mucosubstances increased in the 32 week old fetuses. These results show that the labial glands are histophysiologically differentiated at an earlier stage of development (14 weeks) as compared to lingual glands (20 weeks). However, mucosubstance production would begin during the early phases of embryogenesis for both labial and lingual glands. PMID- 11885245 TI - Bacterial inhibition produced by substances for dentin pretreatment. AB - Dentin treatment before adhesion of composites is performed both to enhance adhesion and to remove the microbial contents of the smear layer. The purpose of these experiments was to evaluate the germicide potential of several dentin treatments used in adhesive systems and of some cleansing solutions. Different germs involved in caries processes were used (Candida Albicans, Streptococcus mutans and Actinomyces naeslundii) to prepare suspensions. Half a milliliter of each of the suspensions was transferred to test tubes and an equal volume of the following substances was added: Scotch Prep Dentin Primer (P), Gluma Cleanser (G), Cleaner Sol. (C), Tubulicid Blue (TB) and Red Label (TR), Blue Experimental Solution (SB) and Red Experimental Solution (SR) and sterile distilled water (control). The preparation was incubated at 37 degrees C for seven days to test viability. P, TR, TB and SB produced complete inhibition of germs tested. The results reveal that, "in vitro", not all the substances tested exert a germicide effect on the microorganisms analyzed. PMID- 11885246 TI - Effect of a toothpaste containing amyloglucosidase and glucose oxidase on recurrent aphthous ulcers. AB - The aim of this study was to assess whether a toothpaste containing amyloglucosidase and glucose oxidase (Z) provoked any effect on minor recurrent aphthous ulcers, (RAU) as compared with a placebo toothpaste (P). Twenty patients (11 females), suffering from minor RAU, participated in this study during a period of 15 weeks. The patients brushed their teeth twice a day with the toothpaste. They were examined once a week to monitor the number and size of ulcers. The mean number of ulcers in both groups was about 40% lower than that found before treatment. Ulcer mean diameter had also decreased in both the placebo (about 32%) and experimental groups (about 66%). There were no statistically significant differences between the two groups in number of weeks with ulcers, in total number of ulcers per patient, and in mean diameter of the ulcers. In conclusion, no significant differences in therapeutic effects could be shown between treatments with Z and P. PMID- 11885247 TI - Scientific presentations and their publication. Experience over a 10-year period in the Argentine Division of the I.A.D.R. AB - At present the "Impact Factor" developed by the Scientific Information Institute (Philadelphia), is an indicator of the quality of the journals in terms of the quality of the papers which are published. The results of odontological research which are presented and discussed at the annual meetings of the DAAIIO are proof of the quality and quantity of scientific production in this area in Argentina. Presentations are undoubtedly numerous. However, their diffusion and qualitative evaluation are of utmost importance. One way to estimate these parameters would be to monitor the ratio between number of presentations and their publication as full papers in journals with a system of referees (Ratio Publ./Pres.). With this aim in mind the presentations at DAAIIO over the 1980-1989 period were considered. Employing the author index of the DAAIIO meetings, we searched for possible publications in the Index Dental and the Index Medicus. The references were compared with the results presented to disregard publications which had not been previously presented at DAAIIO meetings. The data obtained were grouped according to subject area and Research Center. A total of 747 presentations led to 94 publications, the Publ./Pres. Ratio being 1/8 (12.5%). The ratio for each research center was the following: Univ. Buenos Aires 1/7; Natl. Univ. La Plata 1/8; Natl. Univ. Cordoba 1/15. The research groups with the highest publication ratios were Dent. Mat., Natl. Univ. La Plata 1/3; Clinical Pathol., Univ. Bs. As. 1/4; Oral Pathol., Univ. Bs. As. 1/4; Physiol. and Pharmacol., Univ. Bs. As. 1/4; Natl. Univ. Cordoba 1/7. The majority of the publications within 2 years of presentation were in English (69%). These results suggest that an acceptable number of presentations are published, particularly in the area of basic research. We should hope for an increase in the publication/presentation ratio in the future, particularly in the area of applied clinical research since publication would imply technical-scientific quality evaluation of the work by the experts who condition publication. PMID- 11885248 TI - [Dental implants and guided tissue regeneration]. PMID- 11885249 TI - An original orthodontic appliance for experimental mesial movements in rats. AB - The design of an original appliance to achieve mesial movement of the first upper molar on one side in rat is presented. The appliance is constructed in 0.4 mm stainless steel wire forming a parallelepiped 3 mm in width in its anterior sector, 4 mm in width in its posterior sector and 12 mm in length in its lateral branches. In addition a central longitudinal bar is welded to the structure. The lateral branches of the appliance slide freely through the tubes welded to the palatal aspect of the molar bands, cemented in turn to both first molars. A wire 2 mm in length is welded in the anchorage area to the mesial side of the molar tube at 3 mm from the anterior end of the appliance, which acts as a butt. A pre formed nickel-titanium open coil spring 0.23 mm thick, lumen 0.60 mm and 5.00 mm in length is placed distal to the molar tube. The spring is compressed by 1 mm of its original length, the final force being approximately 50 g. The movement achieved was measured on plaster casts obtained from pre and postoperative impressions, and afforded values of 0.250 mm +/- 0.790 mm on the active side and 0.012 +/- 0.011 mm on the passive side. The histologic studies showed an extensive erosive area on the pressure side of the alveolar wall. The appliance presented will be useful to achieve models of experimental movements of only one molar towards mesial with a force of known magnitude. PMID- 11885250 TI - Structural and biochemical modifications in parotid gland induced by fasting. AB - Structural, ultrastructural and biochemical modifications produced by fasting in the parotid gland of guinea pig, were studied. The highest storage of secretory granules was found in the apical cytoplasm after a 12 hour fasting period. The curve of soluble proteins showed that the highest storage of proteins in the parenchyma took place after a 10/12 hour fasting period. Amylase activity reached its highest point after a 10 hour fasting period. We suggest that granules stored in cellular cytoplasm after a 12 hour fasting period would have completed their maturation cycle. PMID- 11885251 TI - Bucodental health condition in patients with Down syndrome of Cordoba City, Argentina. AB - The oral health condition of children and youngsters with Down Syndrome (DS) was evaluated on a sample of 86 mongolic subjects ages 3 to 19, both sexes, residents in the city of Cordoba (Argentina), and compared with control groups. Those persons were attended special educational institutions for the care of that type of disabled individuals. In every age group, the dmf-t and dmf-s indexes were higher in the mongolic children than in the control population, while from the age of 10 onwards the DMF-T and DMF-S of the control population were higher than those of the DS individuals. In spite of this, the scarce participation of the DF component in the mentally disabled showed deficiencies in their dental care. This population exhibited a high frequency of retarded eruption, agenesis, conoidism, Angle's type III malocclusion, posterior cross bite and deficient gingival health. A positive correlation was found between tha activity of Lactobacillus and the amount of Streptococcus and the caries indicators. A high concentration of calcium and secretory IgA was found in the group of mongolic subjects. Our analysis evidences that DS patients are at a disadvantage in relation with healthy individuals in terms of oral health. An early program of preventive measures is proposed (dental hygiene, anti-plaque agents, Therapy of Orofacial Regulation) which would involve the education of parents and teachers. PMID- 11885253 TI - Impairment of molar tooth eruption caused by x-radiation. AB - The effect of x-radiation on erupting molars is presented. New born, 5 day old Wistar rats were locally irradiated in the molar area with doses of 20 Gy. They were killed in two groups, 30 and 60 days postirradiation respectively. Two other groups of non irradiated, age matched rats were killed at the given times. In addition a control group of 5 day old animals was also studied. Radiographic and histologic studies were performed. Odontoblastic atrophy, odontodysplasia, rootless formation, and ankylosis of tooth to bone by osteodentin formation with the resulting lack of tooth eruption were observed. The relation between the histologic alterations and tooth eruption is discussed. PMID- 11885252 TI - Presence of heat shock protein in chronic periapical pathology of pulpal origin. AB - In order to establish whether tissues damaged by Chronic Periapical Pathology (CPP) of endodontic origin produced heat shock protein (HSP) capable of attracting reactive lymphocytes, paraffin sections of samples from the oral cavity of 10 patients with CPP were incubated with commercially available anti HSP monoclonal antibodies. Antibodies were evidenced employing the alkaline phosphatase immunoenzymatic method (DAKO). HSP was found in the lamina propria infiltrated by lymphocytes, in six of the ten samples. These results suggest that HSP may be one of the lymphocyte recruiting factors in the damaged area and opens new possibilities for further research. PMID- 11885255 TI - Prenatal development of human palatine glands: a structural and cytochemical study. AB - The histogenesis of the salivary glands was structurally and cytochemically studied in human embryos and fetuses during the period of 8 to 32 weeks of intrauterine life. Glandular buds appeared at about 12 weeks of embryonic development. The rounded distal ends of the epithelial cords and neighbouring mesenchyma showed small and abundant PAS positive and alcianophilic granules. At age 14 weeks the secretory end pieces and the duct system were seen at different morphologic and structural stages of a differentiation. Mucous acini with scanty mixed acini predominated and serous acini appeared occasionally. From 20 to 24 weeks the mucous acini stained with toluidine blue featured different degrees of metachromasia even in the case of cells of the same acinus. In the ducts it was also possible to identify metachromatic cells intermingled with basophilic cells in the epithelial coat. These findings suggest that the palatine glands present typical histophysiological material from 14 to 20 weeks. The presence of PAS positive, alcianophilic and metachromatic secretory substance in the acinar lumen and the luminal content of ducts suggests that mucin secretion begins during intrauterine life. PMID- 11885254 TI - Comparative effect of antiinflammatory drugs on rat paw edema induced by human sterile dental plaque extract, carrageenan or dextran. AB - The effect of antihistamine (diphenhydramine) or antihistamine and antiserotonin (cyproheptadine) or aspirin-like (acetylsalicylic acid and indomethacin) or corticosteroid (dexamethasone) drugs on the edema induced by various doses of carrageenan, dextran or human sterile dental plaque extract, injected intraplantarily in the rat paw were comparatively studied. The results showed that: (a) human dental plaque extract injected into the rat paw induces a dose dependent inflammatory response, confirming that it is a potent phlogistic agent; (b) the edema induced by the plaque extract though closer to the pattern of carrageenan-induced edema, was different to both the carrageenan- and the dextran induced edema in its time course and the response to antiedema drugs; (c) histamine and serotonin are liberated in the plaque-induced edema but they play no essential role; (d) the inhibitors of arachidonic acid metabolite formation (ASA, indomethacin and dexamethasone) inhibit this inflammation suggesting the presence of prostaglandin-like substances since its first phase. PMID- 11885256 TI - The hamster cheek pouch carcinogenesis model. PMID- 11885257 TI - Dimensional variations induced by local irradiation in cheek skin and oral mucous membrane. AB - The present study was undertaken to characterize and quantitatively analyze an easily reproducible experimental model of rat oral mucosa subjected to a sequence of radiation doses and post-irradiation times using epithelial and connective tissue thickness and cytochrome oxidase activity as end-pints. The radiobiological behaviour of oral mucosa was compared to that of skin subjected to the same experimental conditions. PMID- 11885258 TI - Early steps in bone resorption in experimental periodontitis: a histomorphometric study. AB - A histomorphometric study of the initial steps in the process of bone resorption of inflammatory origin is presented. The study was performed in a model of experimental periodontitis, in which plaque formation is induced by means of a cotton thread ligature placed around the neck of the first molar of Wistar rats. The treated animals were killed in groups after 24, 48, 72 and 96 hours. An equivalent number of untreated rats were employed as controls. After routine histologic processing mesiodistal sections were obtained. The histologic studies showed the sequence of events of the inflammatory process and bone resorption. The histomorphometric studies showed the concomitant increase in erosive areas and number of osteoclasts with the highest values at 72 hours. At 96 hours both parameters decreased. The results are discussed in the light of those afforded by other experimental models, in which bone resorption is induced by orthodontic forces. In these models the increase in the resorptive areas precedes the increase in the number of osteoclasts. The existence of two different mechanisms for the initiation of bone resorption, one which involves the action of an inflammatory environment and the other which results from the action of mechanical forces, is proposed. PMID- 11885259 TI - Germicide effect of several glass ionomer cements. AB - One the most significant characteristics of glass ionomer cements is their ability to release fluoride compounds. This study was carried out to try establish relationships between this property and the possible effect on the growth of microorganisms that are found in carious lesions, Agar BHI medium containing Petri dishes were flooded with strains of Actinomyces naeslundii, Actinomyces israelii and Actinomyces odontolyticus. Cavities were then prepared in the agar and filled with mixtures of several glass ionomer cements. Some of them were polymerizable resin containing products. A zinc phosphate and a zinc oxide-eugenol cement were used as controls. After a seven day incubation at 37 degrees C under anaerobic conditions the inhibition halos around the specimens were measured in a way similar to that used for antibiograms. The statistical analysis of the results showed no significant differences among Actinomyces strains but a significant difference one among cements. Even when no definitive conclusions could be drawn it is worth taking into consideration the effect of glass ionomer cements on microorganisms such as the Actinomyces and continuing studies to establish more clearly what is required from the material to produce a clinically significant outcome. PMID- 11885260 TI - Shear bond strength of adhesive composite systems to human dentin. AB - This study was carried out to evaluate the following adhesive systems under different conditions from the point of view of the bond strength that is obtained on dentin: Scotchbond Multipurpose, 3M: Denthesive II, Kulzer; Optibond, Kerr with techniques # 1 and 5; and A.R.T. Bond. Coltene. Composite resin (Z100, 3M) cylindrical specimens were bonded to flat dentin surfaces and the adhesive strength was determined under shear. No significant differences were found between the group in which products were used with a separate step of dentin demineralization and the group in which this step is not recommended or was omitted. It can be concluded that the use of dentin acid treatment as a separate step is not necessarily required for high composite bond strength even when it could represent a convenient clinical step to prepare enamel surfaces simultaneously. PMID- 11885261 TI - Study of the vascular pattern in oral lichen planus. AB - Oral Lichen Planus is a relatively frequent disease. Its etiopathogenesis is still unknown and it can undergo malignant transformation during its evolution. Thus, data which could contribute to the knowledge of the biology of this disease are particularly significant. The present study involves a quantitative evaluation of the vascular pattern of oral lichen planus. A portion of biopsy specimens taken for histopathologic diagnosis was processed to mark vascular walls using the histoenzymic technique for ATPase activity demonstration. Stained Sections were then evaluated in a semi-automatic magnetic image analyser. The stereologic parameters studied, showed there is no vascular increase in lichen with regard to normal mucosae or leukoplakias, since the number of vascular walls did not show significant differences. Instead, a significant increase was observed in the vascular area. The association of these parameters, indicates that lichen is a more congestive lesion than the other two conditions studied. These findings indicate that the modifications of the vascular pattern could play a role in the etiopathogenesis of oral lichen planus and suggest that the observation of these changes could be a useful element in the histopathologic diagnosis. PMID- 11885262 TI - Suitability of different substrates for reliable bond strength tests. AB - The aim of the present study was to analyze different substrates used to determine composite-dentine bond strength. It comprises 3 parts. In the first, results obtained using bovine and human dentin are compared. In the second, bond strength of composite-bovine dentin sections cut at different angles was studied. In the third part, the effect of different storage solutions on bond strength was analysed. Two methods were used to study the capacity of bovine dentin to reproduce "in vivo" conditions as closely as possible: a) bond strength tests. Tensile strength of composite samples mounted on flat dentin surfaces, treated with bonding agents, stored in artificial saliva at 37 degrees C was measured. b) Observation of exposed surfaces by scanning electron microscopy. Results showed that more reliable values were obtained when using bovine dentin stored in distilled water at 4 degrees C sectioned at an angle of 125 degrees to vestibular face and not further than 7 mm from the incisal edge. PMID- 11885263 TI - Histochemical demonstration of gamma-glutamyl transpeptidase activity in normal rat cartilage and bone. AB - The feasibility of histochemical detection of GGT activity in decalcified bone tissue is proved and the activity distribution pattern of GGT in normal rat cartilage and bone is described. The results suggest the association, in this model, between GGT activity and differentiation mechanisms rather than proliferative processes. The fact that GGT activity in adult tissues which are normally GGT negative has been linked to premalignant transformation confers significance to the study of GGT activity in normal tissues. The results contribute to the knowledge of the biological mechanisms in which GGT activity is involved and to the understanding of the behaviour of tissues which can be used as controls in carcinogenesis models. PMID- 11885264 TI - Thiols in redox mechanism of ribonucleotide reductase. PMID- 11885265 TI - Role of yeast flavin-containing monooxygenase in maintenance of thiol-disulfide redox potential. PMID- 11885266 TI - Identification of cysteine sulfenic acid in AhpC of alkyl hydroperoxide reductase. AB - C165S AhpC in its sulfenate (Cys-SO-) and presumed thiolate (Cys-S-) forms at pH 7 (pKa for sulfenic acid about pH 6.1) exhibit low extinction absorbance bands around 367 and 324 nm, respectively. Sulfenic acid content of the protein can be assessed by its reactivity with the chromophoric TNB anion. Using this technique, H2O2 titrations of C165S AhpC give a maximum of about 1 SOH per subunit on addition of 1.0 to 1.2 equivalents of H2O2. Cys46-SO- is moderately air stable at neutral pH and room temperature and is oxidized at a steady rate of about 10% per half hour. Cys46-SO- of C165S AhpC is reduced in the presence of catalytic amounts of AhpF by approximately 1 equivalent of NADH to regenerate the Cys46-S- species. NBD chloride is extremely useful as a trapping agent for cysteine sulfenic acid. The Cys46-S(O)-NBD adduct absorbs maximally at 347 nm and is 16 amu larger than the Cys46-S-NBD adduct (lambda max = 420 nm) as shown by ESI-MS. Other electrophilic thiol reagents also react with Cys46-SO-; however, iodoacetamide and N-ethylmaleimide reactivities are much lower with Cys46-SO- than with Cys46-S-. These methods are applicable to other sulfenic acid containing proteins, although in some cases the proteins must be denatured in order to provide accessibility of this species toward labeling agents. PMID- 11885267 TI - Glutaredoxins and oxidative stress defense in yeast. PMID- 11885268 TI - Quantitation of protein sulfinic and sulfonic acid, irreversibly oxidized protein cysteine sites in cellular proteins. PMID- 11885269 TI - c-Jun regulation by S-glutathionylation. PMID- 11885270 TI - S-glutathionylation of glyceraldehyde-3-phosphate dehydrogenase: role of thiol oxidation and catalysis by glutaredoxin. AB - The findings in this article illustrate the complexity residing in the regulation of reversible S-glutathionylation of proteins, such as GAPDH. This is clearly reflected in the design of suitable experimental approaches designed to cope with the interaction of several redox-dependent factors. Clear interactions are demonstrated between oxidative modification of GAPDH and its subsequent S glutathionylation. Similarly, a redox interaction between GSSG and GAPDH with Grx as the catalyst is shown, suggesting that the Grx molecule may participate in catalytic S-glutathionylation in intact cells. Furthermore, Grx itself can readily undergo S-glutathionylation, indicating the potential for regulation of this catalyst of the reversible S-glutathionylation of other proteins. The methodologies detailed in this work may provide a good reference point for other attempts to elucidate the mechanism of reversible S-glutathionylation of purified proteins in a manner that more closely resembles the situation arising in intact cells during the generation of oxidative stress. PMID- 11885271 TI - Roles of Nrf2 in activation of antioxidant enzyme genes via antioxidant responsive elements. PMID- 11885272 TI - Enzymatic pathways of beta elimination of chemopreventive selenocysteine Se conjugates. PMID- 11885273 TI - Gene expression and thiol redox state. PMID- 11885274 TI - Tyrosyl radicals and ribonucleotide reductase. PMID- 11885275 TI - Redox flow as an instrument of gene regulation. PMID- 11885276 TI - Optical methods for measuring zinc binding and release, zinc coordination environments in zinc finger proteins, and redox sensitivity and activity of zinc bound thiols. PMID- 11885277 TI - Metallothionein expression and oxidative stress in the brain. PMID- 11885279 TI - Thiol enzymes protecting mitochondria against oxidative damage. PMID- 11885278 TI - Reversible oxidation of HIV-2 protease. PMID- 11885280 TI - Phenylarsine oxide affinity chromatography to identify proteins involved in redox regulation: dithiol-disulfide equilibrium in serine/threonine phosphatase calcineurin. PMID- 11885281 TI - Glutathione reductase from bovine brain. PMID- 11885282 TI - Redox-mediated functional and structural changes in insulin receptor kinase. PMID- 11885283 TI - Redox regulation of protein tyrosine phosphatases by hydrogen peroxide: detecting sulfenic acid intermediates and examining reversible inactivation. PMID- 11885285 TI - Protein cross-linking by self-assisted intermolecular disulfide bond formation. PMID- 11885284 TI - Flavin-dependent sulfhydryl oxidases in protein disulfide bond formation. PMID- 11885286 TI - Sulfhydryl oxidases as factors for mitochondrial biogenesis. PMID- 11885287 TI - Activation of iron regulatory protein-1 by oxidative stress. PMID- 11885288 TI - Mouse astrocyte cultures used to study antioxidant property of metallothionein isoforms. PMID- 11885289 TI - Model peptide substrates and ligands in analysis of action of mammalian protein disulfide-isomerase. PMID- 11885290 TI - Analyzing cotranslational protein folding and disulfide formation by diagonal sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 11885291 TI - Escherichia coli SoxR protein: sensor/transducer of oxidative stress and nitric oxide. PMID- 11885292 TI - Disulfide reduction in major histocompatibility complex class II-restricted antigen processing by interferon-gamma-inducible lysosomal thiol reductase. AB - Constitutively expressed in antigen-presenting cells (APCs), interferon-gamma inducible lysosomal thiol reductase (GILT) catalyzes disulfide bond reduction under acidic conditions. GILT contains a CXXC motif similar to the WCGH/PCK motif of proteins in the thioredoxin family. This class of enzymes catalyzes, at a neutral pH, dithiol oxidation, disulfide bond reduction, and disulfide bond isomerization. A well-established assay spectrophotometrically measures interchain disulfide bond reduction of insulin via the precipitation of aggregating free B chains. However, the insolubility of insulin at low pH limits the use of this assay. To assess the thiol reductase activity of GILT, we employed an assay that uses denatured [125I]F(ab')2 as a substrate, which is detailed in this article. In addition, we discuss approaches used to demonstrate the mechanism of action of GILT and to identify substrates. PMID- 11885293 TI - Thiol oxidation and reduction in major histocompatibility complex class I restricted antigen processing and presentation. PMID- 11885294 TI - Disulfide bond formation in periplasm of Escherichia coli. PMID- 11885295 TI - Protein disulfide isomerase as an enzyme and a chaperone in protein folding. PMID- 11885296 TI - Characterization of redox-active proteins on cell surface. PMID- 11885297 TI - Measurement of reduction of disulfide bonds in plasmin by phosphoglycerate kinase. PMID- 11885298 TI - Redox potential of GSH/GSSG couple: assay and biological significance. PMID- 11885299 TI - Giving children security. Mamie Phipps Clark and the racialization of child psychology. AB - During the 1930s and 1940s, social psychologists became increasingly well-known among progressives battling race prejudice. By the early 1950s, African American psychologist Kenneth Bancroft Clark had become deeply involved with the National Association for the Advancement of Colored People's battle against segregated education in the South. By this time, his wife, who is less well-known in the annals of history, was developing her own reputation as the guiding spirit behind Harlem's Northside Center for Child Development. Her work at the center helped define an increasing interest in the psychology of children of color. This article examines the individual and social contexts of Mamie Phipps Clark's life and argues for greater attention to the dynamics of race and gender in the history of psychology. PMID- 11885300 TI - Kenneth B. Clark in the patterns of American culture. AB - Kenneth B. Clark is most well-remembered as the social scientist cited by the U.S. Supreme Court in footnote 11 of its decision in Brown v. Board of Education in 1954. His presence in that decision came to symbolize the role that social science could play in changing social policy and public attitudes. As an African American social scientist who was prominent during a time of great turmoil over racial issues in the United States, Clark also became a "participant-symbol" in America's discussion of race. Clark contributed to this discussion in the three books he wrote for the general public: Prejudice and Your Child (Clark, 1955), Dark Ghetto (Clark, 1965), and Pathos of Power (Clark, 1974). In this article, the author discusses how these works document Clark's growing pessimism about the prospects for improving race relations. In addition, Clark's place in contemporary American debates about Brown v. Board of Education and the persistence of racial equality is considered. PMID- 11885301 TI - The American Psychological Association's response to Brown v. Board of Education. The case of Kenneth B. Clark. AB - In 1954, in Brown v. Board of Education, the Supreme Court struck down the "separate but equal" doctrine of the Plessy v. Ferguson decision (1896) that was the foundation of school segregation in 17 states and the District of Columbia. Brown is arguably the most important Supreme Court decision of the 20th century in terms of its influence on American history. Moreover, it has a special significance for psychology because it marked the first time that psychological research was cited in a Supreme Court decision and because social science data were seen as paramount in the Court's decision to end school segregation. This article describes psychologist Kenneth B. Clark's role in that case and the response of the American Psychological Association to scientific psychology's moment in a great spotlight. PMID- 11885302 TI - The practice of forensic psychology. A look toward the future in light of the past. AB - In a 1987 American Psychologist article, Tom Grisso summarized the state of forensic psychological assessment, noted its limitations and potential, and offered suggestions for researchers and practitioners interested in contributing to its future. Since that time, there have been many important developments in the field of forensic psychology, as well as in clinical psychology more generally, some of which were anticipated and recommended by Grisso, and some of which were not. Forensic psychology is now at a crossroads, and the specialty must make an effort to respond to current challenges if it is to aid in the administration of justice by assisting legal decision makers. The need to distinguish between and identify levels of forensic knowledge and practice, establish guidelines for practice, educate legal consumers, and devote more attention to treatment issues in forensic contexts is highlighted. PMID- 11885303 TI - The legacy of Kenneth B. Clark to the APA. The Board of Social and Ethical Responsibility for Psychology. AB - The American Psychological Association faced many challenges beginning around 1965, including the challenge to make psychology more inclusive of traditionally underrepresented groups. The larger context of social unrest and public and political focus on social problems framed these challenges. This article describes the events that led to the establishment of the Board of Social and Ethical Responsibility for Psychology in 1972 and highlights the role of Kenneth B. Clark in those events. PMID- 11885304 TI - Are there benefits from NHST? PMID- 11885306 TI - How probable is the null hypothesis? PMID- 11885305 TI - Why chance is a good theory. PMID- 11885307 TI - Beyond objectivity and subjectivity. PMID- 11885308 TI - Fisher's fallacy and NHST's flawed logic. PMID- 11885309 TI - Bayes rules. PMID- 11885310 TI - Part-time dental teaching: a personal retrospective. AB - A prosthodontist in private practice who teaches part-time describes the benefits he has received from combining these aspects of his career. The rewards of teaching early in professional life include professional identification, the opportunity to share views with senior colleagues, and learning from the patterns of mistakes make by students. Later, the rewards included experience synthesizing and organizing one's knowledge and the inspiration to pursue advanced training. Still later, the challenge of working with graduate students who are constantly striving to advance the field prove rewarding. Throughout, the common theme of personal and professional inspiration is a characteristic of dental education. PMID- 11885311 TI - A fresh look at how dental schools prepare dentists for today's practice. AB - The current Vice President for Students of the American Dental Education Association and a recent graduate who also completed an Advanced Education in General Dentistry residency looks at dental education in light of the current demands on beginning dental practitioners. He describes the didactic curriculum that incorporates much new material on the changing foundations of health and dental materials and the comprehensive care, competency-based clinical model typical of most schools. He also discusses how debt, community service, practice opportunities, and initial licensure affect recent graduates' perceptions of the profession. He concludes that continuous learning is the obligation of all dentists, regardless of when they may have graduated. PMID- 11885312 TI - Keeping the curriculum current with research and problem-based learning. AB - Active learning is more effective than passive learning and the critical activities are discovery, mastery, and application. Only 10% of a typical dentist's career is spent in dental school, so the educational experience must provide the tools used for lifelong learning. Problem-Based Learning is a model used in several dental schools that is grounded in these assumptions. PBL makes use of small groups, patient cases serving as the vehicle for learning, with student directed outcomes. PMID- 11885313 TI - Education problems and Web-based teaching: how it impacts dental educators? AB - This article looks at six problems that vex educators and how web-based teaching might help solve them. These problems include: (1) limited access to educational content, (2) need for asynchronous access to educational content, (3) depth and diversity of educational content, (4) training in complex problem solving, (5) promotion of lifelong learning behaviors and (6) achieving excellence in education. The advantages and disadvantage of web-based educational content for each problem are discussed. The article suggests that when a poorly organized course with inaccurate and irrelevant content is placed online, it solves no problems. However some of the above issues can be partially or fully solved by hosting well-constructed teaching modules on the web. This article also reviews the literature investigating the efficacy of off-site education as compared to that provided on-site. The conclusion of this review is that teleconference-based and web-based delivery of educational content can be as effective as traditional classroom-based teaching assuming the technologic problems sometimes associated with delivering teaching content to off-site locations do not interfere in the learning process. A suggested hierarchy for rating and comparing e-learning concepts and methods is presented for consideration. PMID- 11885314 TI - Future of dentistry--education chapter. PMID- 11885315 TI - Overview of issues facing dental education. AB - The Executive Director of the American Association of Dental Education discusses some of the major issues now facing dental education. These include: increasingly complex missions, faculty recruitment and retention, financing, student debt, postdoctoral education, lifelong learning, diversity, clinical education, research, trans-generational learning styles, globalization, and licensure. PMID- 11885316 TI - Requirement-driven dental education and the patient's right to informed consent. AB - In dental education, students spend much of their time treating patients' oral health care needs. Many dental schools still require students to complete a specified number of treatments of various kinds before they can graduate. It often happens that students need to do a particular treatment in order to complete school requirements, when this treatment is not what the patient truly needs, or is not the only treatment indicated for the patient's condition. Consequently, students will be tempted to talk the patient into accepting the procedure. Likewise, educational requirements may tempt the student to postpone certain treatments or forgo non-credit-bearing interventions altogether. We argue that this conflict of interest is inevitable (even though the educational system adopted by the school may mitigate the problem) and analogous to that found in therapeutical experimentation. Hence, we advocate the same ethical solution as has long been adopted for conflicts arising in biomedical experimentation: informed consent. PMID- 11885318 TI - Dental education: one dean's perspective. AB - A dean looks at dental education and the practicing profession from the perspective of three years in dental school administration and sixteen in industry. A significant challenge is to balance costs and standards in the face of well-meaning calls for benefits from those who are not charged with meeting costs. One issue of central importance for education is keeping the curriculum properly positioned in a dynamically evolving profession. The knowledge and skills needed to manage the practice pharmacopeia are used as an example of this problem. It is proposed that schools adopt a future orientation. Dental education must be valued within the higher educational community just as dentistry is valued for its contributions to society at large. Any drift toward proprietary interests must be resisted. This can best be accomplished through a partnership between education and organized dentistry where the lobbying power of the ADA is used to gain the resources education needs and where education, including research and patient care aspects of its mission, are enlisted in support of the practicing community. PMID- 11885319 TI - The value of belonging to your professional organization. PMID- 11885317 TI - A brief history of conflicting ideals in health care. AB - Three medical traditions were in conflict in fifth century B.C. Greece. The Aesculapian view was grounded in illness as a mystery and remedies based on authority. The Cnidian School emphasized early views of disease and science. The School of Cos, commonly associated with the name Hippocrates, was patient and practice based. The history of medicine is a complex intertwining of these traditions. Each developed its own epistemology--theory of how we come to know things and what basis is used to ground truth. These three traditions can be found in dentistry today. Even taken together, however, they fail to account for modern dental practice. PMID- 11885320 TI - Understanding and caring for the child with Asperger syndrome. AB - Nurses in a variety of settings encounter children with the unfamiliar diagnosis of Asperger syndrome (AS). This disorder, which falls clinically along the autism spectrum, is receiving increasing attention because of its inclusion in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) as one of the pervasive developmental disorders. The characteristic features of AS include deficits in social skills, atypical understanding of and use of pragmatic language, behavior problems, and a restricted set of interests. Cognitive abilities vary, and some children with AS have high intelligence. In addition, many children with AS have other conditions, such as attention deficit hyperactivity disorder, Tourette's syndrome, obsessive-compulsive disorder, and depression. The disorder can result in significant functional difficulties in the home, school, and community contexts. A case study highlights the features of AS, and a related individualized school health care plan demonstrates the school nurse's role in family and staff education, monitoring for comorbidities, behavioral management, medication management, support to family members, and referral. PMID- 11885321 TI - Nurse volunteers in school-based hepatitis B immunization programs. AB - School districts across the nation are implementing school-based hepatitis B virus vaccination programs. Because adolescents are at risk of contracting hepatitis B virus, these programs are important in preventing infections. Critical to the success of these programs is having qualified and cost-effective health professionals to administer the vaccine. This article describes the recruitment and training of professional nurse volunteers to administer vaccines in school-based clinics. During the 1998-1999 school year, approximately 60 nurses in Durham, North Carolina, volunteered 300 hours of time to the program. In the first year, the cost of recruiting and training volunteers exceeded the savings from salaried school nurse time. However, savings are expected in future years. Other benefits of recruiting volunteers to administer vaccine include allowing school nurses to remain in their usual assignments, increasing awareness of the health department's mission and school health in the community, and improving collaboration among local health agencies. PMID- 11885322 TI - The nursing outcomes classification: its relevance to school nursing. AB - Two surveys were conducted to determine school nurses' perceptions of the relevance of 190 outcomes developed by the Nursing Outcomes Classification (NOC) research team to school nursing practice. First, a national random survey of members of the National Association of School Nurses was conducted. Participants were asked to identify the percentage of children and families for whom school nurses affect health outcomes. Usable responses were returned from 227 school nurses (22% response rate). Ninety outcomes were perceived as relevant for 30% or more of children and families; 8 outcomes were perceived as relevant for 50% or more of children and families. A second survey--targeting state representatives attending a school nursing leadership meeting--was conducted using a revised version of the instrument. The legend was changed to degree of relevance to school nursing practice, with a response scale of 0 to 10. Usable responses were returned from 31 state representatives (41% response rate). The means of 172 of 190 outcomes were above 5 (median point on the 10-point scale of relevance). The findings of both surveys indicate that a large number of NOC outcomes are useful for documentation of the effectiveness of nursing interventions in school settings. PMID- 11885323 TI - Adolescents and HIV: knowledge, behaviors, influences, and risk perceptions. AB - Although the rate of progression from HIV to AIDS has slowed, the incidence of HIV infection has continued to rise. Many teenagers are knowledgeable about the risks and consequences of HIV, yet a large percentage do not perceive that they are personally at risk. Gaining insight into the perceptions and factors influencing the behavior of teens is critical in HIV and AIDS prevention. A structured 39-item questionnaire was designed to elicit answers that explored 4 areas: knowledge of HIV and AIDS, reported sexual behavior, perceived susceptibility to HIV, and factors influencing behavior. The mean age of the 78 respondents was 15.9 years. One important finding was that 74% of respondents perceived their knowledge of HIV transmission to be "good," yet only 33% were able to answer all of the 8 test questions in this area correctly. In addition, 80% of those who reported engaging in risky behavior such as multiple sexual partners or having sex without condoms also felt they were not personally at risk for HIV. PMID- 11885324 TI - Evaluation of the effectiveness of a youth leadership class in the prevention of depression in adolescents. AB - Depression in adolescence is a common and potentially life-threatening health problem. The purpose of this study was to evaluate the effects of prevention strategies on decreasing the rate of depression in adolescents. A class addressing specific skills identified as having an impact on adolescent depression was taught to youth participating in the program. The nonequivalent control group design was used. The sample consisted of 7th- and 8th-grade students in either the Youth Leadership class or a computer class of a California middle school. The Childhood Depression Inventory was the instrument used. Nine control group subjects and 11 experimental group subjects completed both the pretest and the posttest. The analysis of the data revealed no statistically significant differences between the control and experimental groups. This study should be repeated with larger sample sizes and with greater attention to the timing of pretests and posttests. PMID- 11885325 TI - The facts about faxing. AB - The fax has a vital place in school health nursing practice. Using a facsimile machine can streamline medication communications, parental permissions, and doctor's orders. Nonetheless, there are confidentiality concerns and the potential for abuse. School health office practices for ethical and prudent use of fax technology are outlined. PMID- 11885326 TI - A handbook for student nurses to guide clinical experiences in the school setting. AB - For more than 30 years, nursing students have had the opportunity to have clinical experiences related to their course requirements in the Dallas Public Schools. The Dallas Independent School District School Health Services Department staff provide an orientation to student nurses before their first day in the school clinic. To enhance their learning experience and clarify the regulations and expectations for student nurses, a handbook was prepared for the use of school nurses and the students. The Basic Health Care for the School-age Child: A Handbook for Student Nurses outlines the use of the school as a clinical experience setting. Another purpose for the handbook is to reduce the stress of this clinical rotation for the student nurse and for the staff nurse who serves as the student nurse's preceptor. This article describes the development of the expectations for the clinical experience and the information included in the handbook. An outline of the material included in each section is presented to provide ideas for school nurses who provide or are considering providing a rotation for student nurses in their schools. PMID- 11885327 TI - Pursuing justice in the courts: lessons from the experience of one school nurse. PMID- 11885328 TI - Identifying the mental health needs of preschool children. AB - The city of Chicago offers publicly funded preschool education to 20,000 3- and 4 year-olds through its State Pre-Kindergarten program. The students attend some 300 schools, and their health needs are monitored by 11 nurses and 8 aides. In the last several years, the nursing coordinator recognized the need to improve the mental health assessment skills of the school nurses. To that end, a relationship was developed with a child psychiatric nurse who had expertise in assessing young children's behaviors, particularly in the context of the classroom milieu. The collaboration of the school nurse and mental health nurse consultant was structured as one-on-one sessions, each focusing on a particular child. A case is presented to illustrate the assessment method and accompanying suggestions for early intervention strategies. The case also points out how school nurses can structure assessments of at-risk children that lead to classroom-based interventions. PMID- 11885329 TI - [Comparison of the efficacy of two different iron supplements for anemia prevention in piglets]. AB - In a randomized, confirmatory study performed between July and October 2000 the efficacy of two iron products in preventing iron deficiency anaemia was compared. A total of 102 newborn piglets from ten litters were treated intramuscularly with 200 mg iron as iron dextran per ml, or 200 mg iron as gleptoferron per ml. For true comparison, piglets within a litter of a sow were subdivided into pairs on the basis of birth weight (one pair of the two heaviest piglets, et cetera). Within a pair, treatment with the iron supplements was randomly allocated. One group of piglets was injected at an age of 1 day (experiment 1) and the other group of piglets was injected at an age of 3 days (experiment 2). The piglets were weighed and blood samples were taken at an age of 18 days (experiment 1) or at an age of 19 days (experiment 2). Average daily weight gain and haemoglobin concentrations of both treatment groups were compared. Both products were very effective in preventing anaemia. No significant differences could be found between the two formulations. It can be concluded that iron-dextran and gleptoferron can be used with similar effect for anaemia prevention in piglets. PMID- 11885330 TI - [Modern biotechnology: a new box for Pandora?]. AB - In a bird's-eye view attention has been paid to a number of items of biotechnology. Amongst others to the beginning of the recombinant DNA technology or genetic modification. Mentioned are the bull Herman, the American tomato 'Flavr Savr' and the sheep Dolly. And further something about the supporters and the opponents of genetic modification and the safety of it. The why of transgenic plants and animals. The legislation and the Netherlands as possible transgene production country. PMID- 11885331 TI - [The therapy is worse than the disease]. PMID- 11885332 TI - [Breeding of cattle misjudged by cattle holders]. PMID- 11885333 TI - [Breeders prepare themselves for stricter rules in dog and cat decision 1999]. PMID- 11885335 TI - [Foot and mouth disease control]. PMID- 11885334 TI - [KKM, PBB and MVGK?]. PMID- 11885336 TI - [Predecessors: veterinarians from earlier times (46). Sebastiano Rivolta (1832 1893)]. PMID- 11885337 TI - [New leave policies by activating the work and dependent care law]. PMID- 11885338 TI - Health education: an important role for school nurses. AB - Health education is an important, yet challenging and time-consuming, nursing intervention. It is one of the most important tools school nurses have in teaching students, families, and staff about health. To be effective health educators, nurses need skills in planning and implementing attractive and effective programs to students. They also need to develop skills in evaluating the effectiveness of their efforts to emphasize the impact school nursing can have on the health of children. This editorial highlights the school nurse's role in health education in schools and gives a brief overview of the health education process. Health education provides many opportunities for school nurses to reach out in new and creative ways to students in their quest to promote health and success in the school environment. PMID- 11885339 TI - Vision screening in central Iowa. AB - This study assessed the vision-screening practices of all preschools and elementary schools during the spring of 2000 in the Heartland Area Education Agency (AEA) in central Iowa. Surveys were returned by 7% of the preschools and 56% of the elementary schools. Survey questions were drafted based on recommendations from the Iowa Vision Screening Program Guidelines, which were distributed to all Iowa school districts in 1997. Areas surveyed included vision screening personnel, attainment of students' visual history, rescreening practices, referral and follow-up, and screening procedures. Survey results indicated that there is a need for improvement to standardize vision-screening procedures within Heartland AEA. Time and effort are invested in activities that are not recommended, and not enough effort is being put into recommended activities such as obtaining vision histories, rescreening to avoid overreferrals, and follow-up to make sure students receive required treatment. PMID- 11885340 TI - Excellence in school nursing workshop: Florida's experience in standardizing school nursing orientation and education. AB - Specialty preparation is needed for safe, effective school nursing practice. The Excellence in School Nursing workshop was envisioned as a means to provide a statewide orientation and education program for school nurses in Florida. As a result of a needs assessment, the Florida Departments of Health and Education formed a curriculum development committee to address the educational needs of school nurses. A 3.5-day workshop was designed for school health nursing supervisors and school nurses employed by county health departments, school districts, and community agencies. The workshop was piloted and, based on evaluations and feedback, was revised and replicated nine times throughout the state. The workshop has proved to be an effective method of providing statewide orientation and education for school nurses. The purpose of this article is to describe the workshop curriculum, planning, implementation, and evaluation. Recommendations for adaptation and replication in other states are included. PMID- 11885341 TI - Childhood asthma and indoor allergens: the classroom may be a culprit. AB - Asthma has become the most common chronic illness among children. Indoor environments appear to play a substantial role in the development of asthma. Recent studies indicate strong evidence of a causal relationship between exposure to certain indoor environmental pollutants and development and/or exacerbation of asthma in susceptible individuals. Allergens of concern include those produced by dust mites, cockroaches, cats, dogs, and molds. It is important to better understand this relationship and take preventive and corrective steps to reduce or eliminate these sources in schools, homes, and day care centers. Measures include tracking of asthma and allergic response incidents; monitoring for the presence of allergens and molds; effective cleaning procedures; prompt repair of water leaks and/or moisture problems; control of indoor relative humidity; and proper operation of heating, ventilation, and air conditioning (HVAC) systems. PMID- 11885342 TI - Reduction of illness absenteeism in elementary schools using an alcohol-free instant hand sanitizer. AB - Hand washing is the most effective way to prevent the spread of communicable disease. The purpose of this double-blind, placebo-controlled study was to assess whether an alcohol-free, instant hand sanitizer containing surfactants, allantoin, and benzalkonium chloride could reduce illness absenteeism in a population of 769 elementary school children and serve as an effective alternative when regular soap and water hand washing was not readily available. Prior to the study, students were educated about proper hand washing technique, the importance of hand washing to prevent transmission of germs, and the relationship between germs and illnesses. Children in kindergarten through the 6th grade (ages 5-12) were assigned to the active or placebo hand-sanitizer product and instructed to use the product at scheduled times during the day and as needed after coughing or sneezing. Data on illness absenteeism were tracked. After 5 weeks, students using the active product were 33% less likely to have been absent because of illness when compared with the placebo group. PMID- 11885343 TI - The use of standardized language to describe abdominal pain. AB - The assessment, diagnosis, and management of abdominal pain (AP) in children is a complex and challenging process. The factors that contribute to this complexity are the varied etiologies of AP in children that may be biological and/or psychological, nurses' perceptions of AP, manifestations of AP in children, and the lack of a standardized language to describe AP. In this descriptive study, 64 school nurses completed an investigator-developed survey to determine the usefulness of standardized language in describing AP. This study identified the most frequently used assessments to document complaints of AP; activities from the Nursing Interventions Classification "pain management" intervention used to treat AP; and relevant indicators used by school nurses from the Nursing Outcomes Classification "pain level" and "symptom severity" outcomes. Use of standardized language to document nursing care will make nursing's contribution to the care of children with AP more visible and provide data that will identify and monitor trends of AP in the school setting. PMID- 11885344 TI - The facts about e-mail. AB - The use of E-mail in the school setting has become a standard method of communication. E-mail can increase the efficiency of communicating with parents and students. Privacy and security considerations regarding the use of E-mail to transmit health information must be addressed. This article outlines regulations and the best practices for using E-mail in the school health office. PMID- 11885345 TI - Using nicotine replacement therapy in treating nicotine addiction in adolescents. AB - Cigarette smoking is the greatest cause of preventable death and disability in the United States. More than 3,000 children in the United States begin smoking each day. Smokers experience withdrawal symptoms that can be ameliorated by pharmacological interventions. These interventions include Zyban (Bupropion HCl), Nicorette gum, Habitrol patch, Nicoderm patch, Nicotrol inhaler, and Nicotrol NS spray, along with their generic counterparts. This article reviews each of these agents, the time course of nicotine withdrawal symptoms, and the Fagerstrom Tolerance Questionnaire and presents a framework for assisting the nicotine addicted student in smoking cessation. PMID- 11885346 TI - A patient pathway to renal replacement therapy: implementation in the progressive renal insufficiency population. AB - Care paths and patient pathways are collaborative interdisciplinary tools developed to provide quality care. One goal in progressive renal insufficiency (PRI) care is to provide patients and family members with relevant and timely information to help them cope with medical therapy and lifestyle changes required to slow the progression of renal insufficiency. At Halton Healthcare Services, clinical and patient pathways have been widely used with success. Recognizing the potential benefits of a patient pathway for the PRI population, our interdisciplinary team developed a pathway that outlines the journey to renal replacement therapy. The pathway provides the patient with some insight into the future, as their disease process continues to have impact on their life. Reviewed by a patient within our PRI program during the development process, we have tailored this pathway to meet the needs of this unique population. We predict this patient pathway will have positive impact on the patient's understanding of the role of the PRI clinic and the partnership between the team members, the patient, and the family. PMID- 11885347 TI - An educational tool for the care of the hemodialysis patient admitted to an acute care hospital. AB - The goal of this article was to develop an educational tool for nurses working in an acute care hospital in the hope of improving quality of care for hemodialysis patients. In Canada, the prevalence of end stage renal disease is increasing by approximately 10% annually (Mendelssohn et al., 1999). Today, the hemodialysis patient is often older than previously, and may well have multiple comorbidities resulting in hospital admissions in a multitude of settings. Hemodialysis patients are not always admitted under the nephrology service due to lack of beds or medical necessity. For example, a hemodialysis patient with an acute myocardial infarction would be admitted to a cardiology unit, where the cardiology staff are better able to treat the patient's primary diagnosis. When patients are transferred among the various hospital units, effective communication of knowledge and expertise between nursing staff is essential for care of the hemodialysis patient to be seamless. PMID- 11885348 TI - Continuous quality improvement: at the grassroots. AB - As a philosophy and a belief system, continuous quality improvement (CQI) provides the basis for organizational commitment to strategic leadership, customer focus, an empowered workforce, and reliance on evidence to improve processes and outcomes. This paper describes the process engaged in by the hemodialysis unit at St. Joseph's Health Care, London to formalize our CQI processes. The paper emphasizes: the selection of outcome measures, the strategies to link outcomes to improvement opportunities, and the resulting improvement projects. The formal commitment to CQI has affected both patient care and staff morale in many positive ways. PMID- 11885349 TI - What are professional standards and how do I use them in my practice as a nephrology nurse? PMID- 11885350 TI - Keep it simple: teaching totally blind patients using Baxter's Twin Bag peritoneal dialysis system. PMID- 11885351 TI - Protothecosis in human medicine. AB - Prototheca spp. are ubiquitous achlorophyllous algae that produce disease in humans and animals. In the past years infections with Prototheca have obtained increasing importance in human medicine. The cases have been classified into three clinical forms: cutaneous and/or subcutaneous infection, synovitis of olecranon bursa or other fibrous tissue and systemic infection. Patients with a mild degree of immunosuppression may become colonized by Prototheca spp. with a subsequent worsening of their immune surveillance and spread of the disease. Among the numerous pharmacologic agents tried, amphotericin B is the most promising. Successful treatment of protothecosis involves radical excision of the involved structures. PMID- 11885352 TI - A pilot study of three methods for the reduction of bacterial contamination of dental unit water systems in routine use. AB - Three different methods for minimizing the bacterial contamination of the water system in a SIRONA C2 type dental unit were investigated sequentially. Without any decontamination method, water from the hand piece, air-water-jet and mouthwash were continuously contaminated by 10(3) to 10(5) colony forming units (cfu) of aerobic mesophilic bacteria per milliliter. A reduction to below 100 cfu/ml was achieved by continuous adding of a chemical microbicide based on hydrogen peroxide and silver ions. However, this was only possible after rinsing the system thoroughly for at least two minutes after interruptions of the treatment. Long-lasting low counts of below 100/ml were obtained by means of an in-line bacteria filter, in connection with the provision of a thermo-chemical or thermal decontamination of the water pipes and hand pieces after the filter. The electrolyte release of chlorine from the dental unit tap water by anodic oxidation without addition of any chemical disinfectant also resulted in continuously low colony numbers of the water. In this case, regular decontamination of the end parts of the pipes and hand pieces was not necessary. PMID- 11885353 TI - Microbicidal efficacy of superheated steam. II. Studies involving E. faecium and spores of B. xerothermodurans and B. coagulans. AB - The authors' studies on the heat resistance of microorganisms and the superheating of steam (Spicher et al., Zbl. Hyg. Umweltmed. 201, 541-553, 1999) have been continued, using the vegetative bacterium, Enterococcus faecium, and spores of B. xerothermodurans and B. coagulans. The temperature of the saturated steam was 68 degrees C (E. faecium), 105 degrees C (B. xerothermodurans) and 110 degrees C (B. coagulans), respectively. The steam was superheated by 30-40 K, as a maximum. The test organisms had been fixed to fibre glass fleece. The time of exposure to saturated or superheated steam after which 50% of the bioindicators yielded no viable germs was used as a measure of resistance of the organisms. In the discussion, reference has also been made to the findings for B. subtilis and B. stearothermophilus spores obtained in the preceding study. E. faecium exhibited a maximum resistance when superheating the steam by 5 K. The superheated steam required a 74-fold exposure time compared to saturated steam of 68 degrees C. Resistance became gradually reduced as superheating was further increased. Even superheating by 30 K required a 11-fold exposure time. The bacterial spores exhibited maxima of resistance on superheating by 23-30 K. In these cases necessary exposure times were 119 (B. subtilis), 30 (B. xerothermodurans), and 4 times (B. coagulans, B. stearothermophilus) longer than those required in saturated steam. In the superheating range below 10 K, behaviour patterns varied. Thus, heat resistance may initially become reduced with increasing superheating (B. coagulans), remain on almost the same level as under exposure to saturated steam (B. stearothermophilus), reach a stage of weakly enhanced resistance (B. xerothermodurans), or approach a maximum of resistance in an almost linear mode (B. subtilis). It appears that there are differences between strains of one and the same bacterial species. PMID- 11885354 TI - Leukemia mortality and occupational exposure to rubber: a nested case-control study. AB - OBJECTIVE: The excess risk of leukemia in rubber industry was considered to be real and attributed to the exposure to solvents, particularly benzene. METHODS: Following a nested case-control study, we used the data of 7 leukemia deaths in 1973-1997 and 28 controls matched for sex and age from the same cohort of a rubber plant. Leukemia risks due to exposure to rubber were assessed, unadjusted and adjusted for non-occupational factors by conditional logistic regression. RESULTS: OR for leukemia was found to be 7.81 (95% CI = 0.77-78) in grouped analysis for one or more years of work in the inner tire tube department. The models for continuous exposure variables indicated that working for one year in the inner tire tube department was associated with a 10 percent increment in the OR (95% CI = 1.00-1.24; score test for linear trend: chi 1(2) = 6.27, P = 0.012). The confounding effects studied could not be ruled out for the excess risks. No excess risk was found in the remaining four departments. CONCLUSIONS: Because of widespread exposure to various carcinogens in the process of making inner tire tubes, removal of a single agent (benzene) may not eliminate the risk of leukemia in the entire industry. PMID- 11885355 TI - In vitro effects of incinerator fly ash on pulmonary macrophages and epithelial cells. AB - Fly ash from a municipal waste incinerator was used as a model for atmospheric particles in order to identify parameters relevant for the induction of adverse health effects. The aim of this study was to compare the biological effects of the total incinerator fly ash (IFA), the soluble and the insoluble fraction with the effects of quartz by in vitro toxicity studies. The previously sized fly ash (< 20 microns) was characterized by elemental composition and particle size distribution. The particles were administered to rat alveolar macrophages (NR8383) and human bronchial epithelial cells (BEAS-2B) at different amounts via the medium. The total IFA and its insoluble fraction were shown to induce cytotoxicity and cytokine release in a dose-dependent manner. The soluble fraction was nearly unable to induce cytotoxicity and TNF-alpha release but showed potent induction of IL-8 release in BEAS-2B cells at increasing concentrations. Quartz caused similar effects compared to IFA in NR8383 but was less effective in BEAS-2B. PMID- 11885356 TI - Mineral composition other than quartz is a critical determinant of the particle inflammatory potential. AB - The aim of this study was to examine the inflammatory potential of stone quarry particles with differing mineral and metal composition and if the effects could be related to the leaching of metals from the particles and if antioxidants would reduce the cytokine release. After intratracheal instillation of rats with a type of mylonite (median size 8 microns) we found a stronger inflammatory potential of mylonite than of quartz at 20 h after treatment. In isolated rat type 2 cells and human epithelial lung cells (A549) mylonite induced a much greater release of MIP 2/IL-8 than quartz or a type of basalt and a feldspar. The mylonite particles were more potent even when compared to smaller size fractions of quartz. Thus mineral composition can be more important than size in eliciting acute inflammatory responses. The content of metals in basalt and mylonite showed minor variations with somewhat more metals present in basalt. The release of metals from the two particle types varied, but in general more metals were released from basalt than from mylonite particles. However, metal release was not related to the differences in proinflammatory effect. Antioxidants seemed to decrease the release of cytokines induced by mylonite particles, but a suppression of basal cytokine release by antioxidants was also observed, questioning the involvement of oxygen radicals in the mylonite-induced effects. PMID- 11885357 TI - Biomonitoring of polycyclic aromatic compounds in the urine of mining workers occupationally exposed to diesel exhaust. AB - Diesel exhaust is considered a probable human carcinogen by the IARC. Biomonitoring of workers occupationally exposed to diesel exhaust was performed to determine their internal burden of diesel associated aromatic compounds. Personal air sampling also allowed to determine the exposure of the miners at their work place towards several polycyclic aromatic hydrocarbons (PAH) and nitro arenes, the latter of which are thought to be specific constituents of diesel exhaust. For biomonitoring the urine of 18 underground salt miners was collected during and after their shift for 24-hours. half of the 18 miners were smokers. The urinary levels of 1-hydroxypyrene and hydroxylated phenanthrene metabolites were determined as biomarkers of PAH exposure, whereas urinary levels of some aromatic amines were chosen to monitor exposure towards specific nitro-arenes from diesel exhaust like 1-nitropyrene and 3-nitrobenzanthrone and to monitor the human burden by these compounds from inhaled cigarette smoke. Non-smoking workers exposed to diesel exhaust excrete an average level of about 4 micrograms phenanthrene metabolites, whereas the urinary levels in smokers were up to 3-fold higher. In summary the results indicate that (i) diesel exposure led to an increase of PAH metabolism in the workers examined, most probably by an induction of cytochrome P450 (ii) smokers could be identified in accordance with earlier studies by their increased ratio of phenanthrene metabolites derived from 1,2- and 3,4-oxidation and their higher amounts of excreted 1-naphthylamine, and (iii) the excreted amounts of aromatic amines found as metabolites of the nitro-arenes were about 5- to 10-fold higher as one might expect from the levels determined by personal air sampling at the workplace of the individuals. PMID- 11885358 TI - An interdisciplinary therapeutic approach for dealing with patients attributing chronic fatigue and functional memory disorders to environmental poisoning--a pilot study. AB - Nonspecific symptoms and a general feeling of ill health that is difficult to objectify are the commonest health problems with which patients present to an Environmental Medicine Outpatient Department (OPD). Of this group, a great proportion meets the classification criteria for Chronic Fatigue Syndrome (CFS) or Functional Memory Disorders in association with Idiopathic Chronic Fatigue (FMD-ICF). This is a longitudinal study of the OPD of Environmental Medicine, Freiburg University Hospital, Germany, to determine the feasibility and impact of an interdisciplinary therapeutic approach (self-help program, acupuncture, psychosomatic support by group interventions) in 8 patients with CFS, FMD-ICF, or CFS in association with self-reported Multiple Chemical Sensitivities (sr-MCS). The intervention took into consideration the patients' need for treatment of physical aspects of their disease. This is an important step to motivate patients into required psychosomatic support. Although none of the patients was willing to accept psychosomatic support or psychotherapy at study outset, acceptance of psychosomatic group interventions was high during the study course. Additionally five patients started with personal counseling at the Psychosomatic Clinic, and, without feeling stigmatized, 4 patients started with specific psychotherapy. The patients' quality of life showed no increase after four months, but, as shown by the Sum-Score of SF-36, it had improved significantly at the end of the study, which covered eight months' treatment (p = 0.015). Two follow-up investigations showed that this improvement probably persisted in part (mainly in the dimensions mental health, social function, physical role function, and vitality). In conclusion our interdisciplinary therapeutic approach indicates successful treatment of patients attributing CFS, CFS/sr-MCS, and FMD-ICF to environmental poisoning. We now plan to conduct a randomized controlled trial in the future. PMID- 11885359 TI - Role of increased environmental Aspergillus exposure for patients with chronic obstructive pulmonary disease (COPD) treated with corticosteroids in an intensive care unit. AB - We report about a 75-year-old woman and a 62-year-old man hospitalised for infection-related exacerbation of chronic obstructive pulmonary disease (COPD). In both patients, respiratory function worsened after initial stabilisation and the disease took a fatal course. A careful inspection of the intensive care unit (ICU) revealed several circumstances known to be risk factors for invasive aspergillosis (reconstruction activities near to the ICU, contamination of the window sills with pigeon droppings, moist building materials due to water leakage). The case reports suggest that both critically ill patients receiving high dose corticosteroid medication possibly have acquired aspergillosis on account of increased environmental exposure to Aspergillus conidia (> 10(2) CFU/m3 air). However, due to the severity of the disease confirmation by invasive diagnostic procedures was not possible. The role of high dose corticosteroid treatment as a risk factor for invasive aspergillosis should be taken into consideration, and increased exposure to fungi consequently be reduced in health care environments, especially for patients at risk. PMID- 11885360 TI - Uterine molecular responses to bisphenol A treatment before and after decidual induction in pseudopregnant rats. AB - The results of the study demonstrate that the weak estrogenic action of bisphenol A (a daily subcutaneous dose of 200 mg/kg on 4 consecutive days, administered before or after decidual induction that occurs on day 4 of pseudopregnancy) on deciduoma growth in pseudopregnant Sprague-Dawley rats, was functionally associated with the hormonal status of the uterus. Whereas bisphenol A displayed uterotrophic action during the pre-decidual, estrogen related period, it inhibited decidual growth and progesterone secretion during the post-decidual, progesterone-dominated period. The estrogenic action of bisphenol A on uterine decidual growth was not correlated with the reduced levels of estrogen receptor binding sites and mRNA expression, nor the unchanged serum estradiol concentrations. BPA action appeared to be antagonized by progesterone. PMID- 11885361 TI - Dietary intake of persistent organic pollutants (POPs) by German children using duplicate portion sampling. AB - The dietary intake of persistent organic pollutants (POPs) was studied in 14 German children at the age of 1.5 to 5.3 years from the North Sea island Amrum. A total of 98 duplicate samples were collected between April and May 1995. The sampling period for each participant was 7 days. Concentrations of POPs were measured by capillary gas chromatography and electron capture detection. The median daily intake [ng/(kgbw.day)] for the different compounds were as follows: 1.9 for alpha-HCH, 1.2 for beta-HCH, 9.5 for gamma-HCH, 4.6 for HCB, 3.4 for DDT, 11.2 for DDE, 1.1 for PCB 101, 5.1 for, PCB 138, 5.2 for PCB 153 and 2.2 for PCB 180. Compared to acceptable or tolerable daily intake (ADI/TDI) proposed by WHO and other organizations the dietary intake of POPs was low. The median values of the ADI/TDI for the POPs was less than 2.2%. The highest percentage of tolerable intake was found for the sum of PCB and amounted to 20.4%. However, compared with minimal risk levels (ATSDR), the percentage of dietary intake was much higher, especially for gamma-HCH (based on median intake: 95%), for HCB (23.1%) and for PCB (69%). PMID- 11885362 TI - Increased incidence of allergic sensitisation and respiratory diseases due to mould exposure: results of the Leipzig Allergy Risk children Study (LARS). AB - To investigate mould effects on health, the concentration of mould spores in air and dust was determined during a prospective cohort study. Clinical outcome was estimated by questionnaires and determination of specific IgE antibodies and intracellular cytokine production of T cells. A significant association was observed between the incidence of respiratory tract infections and exposure to Penicillium spores. Moreover, Aspergillus exposure was found to be associated with allergic rhinitis or related symptoms. In addition, T cells of children exposed to Aspergillus showed a significantly lower content of TH1 cytokines (IFN gamma, TNF-alpha, IL-2) producing cells. Our data suggest that mould exposure is associated with several effects on health, depending on the species involved. PMID- 11885363 TI - Time courses of sensory irritations due to 2-butanone and ethyl benzene exposure: influences of self-reported multiple chemical sensitivity (sMCS). AB - The individually different effects of exposure to comparable levels of chemicals might be partly explained by dissimilar response sensitivity towards chemicals. Multiple chemical sensitivity (MCS) might be the clinical endpoint of this altered sensitivity. Concerning a subclinical range of chemical sensitivity, 'challenge studies' with people reporting chemical sensitivity are needed to improve the knowledge about such differences. The chemical and general environmental sensitivity questionnaire (CGES) is a standardized screening tool for the selection of this group. In the present study 24 healthy male volunteers, half of them classified as sMCS-subjects, were experimentally exposed to 2 butanone and ethyl benzene at different levels (TLV-level vs. odor threshold). The strength of self-reported sensory irritations (nasal and ocular) and symptoms of bad smell were assessed, prior, during, and after the 4 hours of exposure. The time courses of sensory irritations were affected by sMCS. Across all exposure periods sMCS-subjects showed increasing symptom scores while control-subjects did not. Symptoms of bad smell were affected by three exposure-related factors (substance, level, duration) without any additional influence from the sMCS factor. Starting from these results it could be concluded that the time-depending influence of reported chemical sensitivity is most prominent for subjective data of sensory irritations. PMID- 11885364 TI - Psychophysiological functions of subjects with self-reported multiple chemical sensitivity (sMCS) during experimental solvent exposure. AB - The study examined the assumption of a higher sensitivity of autonomic functions of subjects with self-reported multiple chemical sensitivity (sMCS) during environmental exposure. The hypothesis was tested in a laboratory study with standardized exposures. Twelve healthy male subjects (26.4 +/- 5.4 y) with and 12 male control subjects (25.7 +/- 3.8 y) without self-reported multiple chemical sensitivity (sMCS), selected by a questionnaire, were included in the experimental study. At four different days the subjects were exposed in a random order to solvents for four hours: 10 ppm or 98 ppm ethyl benzene, 10 ppm or 189 ppm 2-butanone. Heart rate and breathing rate were analysed for two 30-minutes periods of vigilance testing at the beginning and end of exposure. In sMCS subjects both functions were elevated at the beginning of the testing periods with a tendency to decrease over the 30-minutes periods. Control subjects revealed a relatively constant level (breathing rate) and a small increase (heart rate) during the periods. These group differences were obvious for all experimental conditions across substances and levels of exposures. Furthermore, the mean of the breathing rate of sMCS-subjects was generally higher compared to the control subjects. While the assumption of a generally altered sensitivity of autonomic functions of sMCS-subjects to environmental changes seems to be supported, no specific reactions to the type or level of the chemical exposure were found. PMID- 11885365 TI - Sorption of toxic heavy metals to soil. AB - The surface soil is a major recipient of pollutants, including heavy metals, through atmospheric deposition, agricultural practices, and waste disposal. In the present work the sorption capacity of different types of soils to toxic heavy metals, i.e. chromium, copper, cadmium and lead has been studied. Experimental adsorption data for metals to the soil obtained by the batch method were fitted by linear isotherm. The various soils showed a very different behaviour in sorption of heavy metals. The distribution coefficient Kd, which is an indication of the adsorbing capacity of the substrate, varies within a wide range, from 57 to 53,000 l kg-1. Desorption of metals from the solid phase was found to be small, indicating that the soil matrix is affecting the metal mobility by modifying the bonding of pollutants to the soil system consequently affecting the potential for soil remediation processes. PMID- 11885366 TI - Soluble intercellular adhesion molecules in the serum of subjects exposed to dust at different workplaces--correlation to airway symptoms, lung function, tobacco and dust exposure. AB - Soluble intercellular adhesion molecule 1 (s-ICAM-1) was evaluated as biomarker indicating inflammatory processes in the airways of persons exposed to salt dust, ceramic dust and flour dust. ICAM values in the serum of these workers were related to airway symptoms, lung function (FEV1), inhalable dust dose and tobacco consumption. A weak relation was found to airway symptoms (cough and phlegm) and FEV1. Consistent elevated ICAM levels in smokers suggest, that ICAM indicates inflammatory processes following strong irritants such as tobacco smoke. In contrast to lung function analysis, serum ICAM does not support inflammatory changes by salt dust or ceramic dust. PMID- 11885367 TI - Interference of CD95 expression on human lymphocytes. AB - The study presents the exogenous influence of cadmium in comparison with zinc on the apoptosis of human lymphocytes by CD95 expression and its kinetic changes. The salts of both metals were used in final concentrations of 20 microM in cell cultures with whole blood. The duration of cultivation was 18 and 90 hours. The expression of surface antigens was evaluated by flow cytometry with monoclonal antibodies. In cultures of not stimulated cells we found in average 51.54% CD95 positive lymphocytes. The kinetic study of untreated cells showed elevation after 18 hours of cultivation and a very low expression after 90 hours. The CD95 expression on lymphocytes in cell culture with cadmium and zinc was lower after 18 hours of cultivation than in untreated cells. After 90 hours cultivation we found low levels of CD95 expression on cells treated with cadmium and a great individual variability in the number of positive cells upon the influence of zinc. PMID- 11885368 TI - [Acute pancreatitis after surgical treatment of abdominal aortic aneurysm]. AB - INTRODUCTION: Acute pancreatitis after surgical treatment of non ruptured aneurysm of abdominal aorta is a rare complication, considered to be due to pancreatic ischemia or peroperative trauma of pancreas. The aim of this study is to describe 4 new cases of this complication and to discuss its etiology. PATIENTS AND METHODS: From January 1995 to November 2000, 365 patients underwent elective surgery for a non ruptured abdominal aorta aneurysm. Four (1.1%) men, aged 66 to 79 years and operated for an aneurysm which diameter ranged from 60 to 77 mm, developed postoperative acute pancreatitis. The abdominal approach was a midline incision in 3 cases and a retroperitoneal lombotomy in one case. Superior pole of the aneurysm always adjoined or involved the right renal artery. The aortic clamping was supra-renal in 3 cases and celiac in one case. Diagnosis of acute pancreatitis was established at days 2, 4, 12, and 23 after surgery on abdominal computed tomography in 3 cases and at reoperation in one case. RESULTS: Three patients died, including 2 from early multiple organ failure and one peroperatively during surgical attempt to treat a prostheto-digestive fistula. One patient was alive and asymptomatic with a 2-years follow-up. CONCLUSION: Acute pancreatitis is a rare and serious complication after surgical treatment of abdominal aorta aneurysm. Its diagnosis is often delayed. The main etiological factor of this complication could be trauma of pancreas during supra-renal clamping through a midline incision. PMID- 11885369 TI - [Evaluation of laparoscopic bariatric surgery using the BAROS score]. AB - AIM OF THE STUDY: Mid-term assessment of laparoscopic adjustable silicon gastric banding (Lap-Band) by a specific score. PATIENTS AND METHODS: One hundred consecutive patients received by mail 12 to 54 months after laparoscopic adjustable gastric banding a questionnaire including the Bariatric Analysis and Reporting Outcome System (BAROS) which is the only specific and validated instrument for measuring the quality of life after bariatric surgery. This score includes five categories of results (failure, fair, good, very good, excellent). It uses three major fields: the quality of life, excess weight loss, and medical comorbidities evaluation. RESULTS: Seventy three patients answered back with a mean follow up of 24.6 +/- 10 months. Forty six (2/3) had lost more than 50% of their weight excess. Sixty six experienced an improvement of their medical conditions following surgery. Final results were good or excellent for 60 patients (82% of those who answered back). Failure was reported in 7 patients (2 "sweet eaters" and 2 pouch dilatations) which needed a surgical treatment. CONCLUSION: This evaluation based on the BAROS confirms its validation in France and the good mid-term results of bariatric surgery based on the Lap-Band. PMID- 11885370 TI - [Is subtotal bilateral thyroidectomy still indicated in patients with Grave's disease?]. AB - AIM OF THE STUDY: To evaluate the morbidity and the functional results of subtotal bilateral thyroidectomy in patients (TST) with Graves' disease. PATIENTS AND METHOD: A retrospective study was performed in 128 patients. They were 23 males and 105 females with a median age of 34 years (range: 14-68). Weight of remnant tissue was between 4 and 5 g. Thyroid functional status was evaluated, at 3 months and after a follow-up period ranged from 1 to 5 years, by measurement of serum concentration of free T4 and/or free T3 and TSH. RESULTS: They were no post operative death. Surgical complications were 2 vocal cord palsies and 17 hypocalcemia (inf. to 2 mmol/L). After a median follow-up of 2 years, they were no longer any cases of vocal cord dysfunction and no case of permanent hypoparathyroidism. Functional results were established in 118 patients: 46 patients had clinical hypothyroidism (39%), 64 patients had latent hypothyroidism or euthyroidism (54.2%), and 8 had recurrent hyperthyroidism (6.8%). CONCLUSION: These results suggest that TST with a remnant mass inferior to 5 g provides a low level of recurrent hyperthyroidism and allows to give no drug therapy to half patients. In our opinion, TST is still indicated in Graves' disease. PMID- 11885371 TI - [Outpatient laparoscopic cholecystectomy]. AB - OUTPATIENT LAPAROSCOPIC CHOLECYSTECTOMY: The laparoscopic technique is the procedure of choice for cholecystectomy. This procedure is done on ambulatory setting in the United States and Europe but no experience was reported in France. AIM OF THE STUDY: To report the organisation and results of our initial 100 consecutive patients operated for a laparoscopic cholecystectomy on an outpatient basis. PATIENTS AND METHODS: After assessment of the prevention of pain and nausea or vomiting after laparoscopic cholecystectomy on hospitalized patients, a prospective trial was done on our first 100 patients for outpatient laparoscopic cholecystectomy on routine basis. RESULTS: During the period, 27.4% of patients were entered on an ambulatory basis. 72% of patients did not need any medication post-operatively in the structure. 17 patients were admitted: in five cases, decision was done pre-operatively, one patient went back home against medical advising; in three cases, peroperatively, and in 10 cases postoperatively. Four patients were readmitted between the fifth and sixteenth post-operatoire day. CONCLUSION: An adequate organisation for day case surgery, a good selection of patients on medical, surgical and environmental criteria, simple procedures to prevent pain or nausea vomiting post-operatively allow use to assert that hospitalisation is unjustified for laparoscopic cholecystectomy in a quater of patients. PMID- 11885372 TI - [Discordance between actual and radiologically estimated size of an incidentaloma. Delay in questions regarding operative decision-making criteria]. AB - AIM OF THE STUDY: The accurate assessment of tumour size is an important consideration during preoperative evaluation of adrenal tumors, particularly incidentaloma; however the "size criteria" is still a controversial topic in some respects: size is a bad indicator of malignancy, there is still a confusion in the "grey zone" for tumors between 3 and 6 cm, and no universal consensus on the exact cut-off value for resection has been agreed. Nowadays it is clearly accepted that the "size criteria" alone is extremely limited in the assessment of adrenal tumor, moreover some studies suggested the relative inaccuracy of conventional CT in evaluating the size: radiological examination underestimated consistently adrenal tumor size. The aim of this study was to confirm those suggested data. PATIENTS AND METHODS: Our study compared the radiological estimated size and the histological size of 26 incidentaloma operated on with a laparoscopic approach. RESULTS: Our data confirm the inaccuracy of CT and MRI in predicting the size of incidentaloma particularly for tumor measuring less than 3 cm. CT and MRI significantly underestimated size of adrenal tumors, 108% for MRI and 101% for CT-scan. CONCLUSION: The decision to operate, even with the advent and safety of laparoscopic adrenalectomy, cannot only rely on the "size criteria". Radiologists have to perform multiple 1 mm cuts until the very superior and inferior tip of this tumor in order to provide a better estimation of the size. PMID- 11885373 TI - [Translaryngeal tracheostomy using the Fantoni technique: report of 104 cases]. AB - OBJECTIVE: Surgical tracheostomy morbidity led the authors to investigate new election techniques. The aim of this retrospective study was to assess the translaryngeal tracheostomy (TLT), complications and cost. METHODS: From January 1998 to January 2001, 104 patients were treated with TLT modified: 69 males (66.3%) and 35 females (33.7%), average age 52.6 +/- 9.5 years. The original pathologies were: traumatical (36), neurological (37), surgical (9), heart (4), respiratory (18). The average time between intubation and execution of TLT was 4.2 +/- 1.3 days. RESULTS: Fifty four patients died (52%) and 50 patients lived (48%). Two complications (1.9%) occurred in those who survived: a breaking of the guidewire in traction. Extraction of the tracheostomy tube by clamp, a haemorrhage in 2nd post-operative day due to a thyroid vessel lesion. The haemostasis was performed by classical tracheostomy. The average number of days to decannulation was 25 +/- 1 days. CONCLUSIONS: TLT reduces trauma or trachea and neighbouring structures. This technique is safe and easy. TLT is an effective method, in non-urgent situations, in children and adults, as well as in brachytypes and the obese. PMID- 11885374 TI - [Cowden's disease in an adolescent]. AB - Cowden's disease is an autosomal dominantly inherited syndrome characterized by mucocutaneous lesions and multiple hamartomas. We report here a 12 years-old boy case with craniomegally, intestinal polyps, epilepsy and multiadenomatous goiter. All the lesions were beginnings. The predisposing genetic defect has been assignated to chromosomal 10 (PTEN-gene mutation). A long term follow-up is necessary because of the risk of malignancies. PMID- 11885375 TI - [Iliac venous leiomyosarcoma revealed by cruralgia: a case report]. AB - The iliac venous leimyosarcoma is rare, usually malignant, and often occurs with oedema or phlebitis. We report one case of iliac venous leiomyosarcoma revealed by cruralgia. A 69 years old patient, presented with a left cruralgia which had been developing for three months and which happened after an insignificant trauma. The clinical examination objectified a stiff painful mass of the left iliac fossa together with left psoitis. The initial pelvic tomodensitometry showed a mass at the contact of the psoas muscle. At first, the diagnosis of a psoas haematoma complicated by a compressive cruralgia was evocated. Two months ago, the patient had a pulmonary embolism. At his hospitalisation, considering the persistent cruralgia, a tomodensitometry and a pelvic magnetic resonance imaging were carried out and had shown an heterogeneous mass that was including the iliac vessels. The result of the anatomopathologic examination was leiomysarcoma. Due to the disease's evolution (pulmonary metastasis), only a medical treatment by chemotherapy was undertaken and the patient died a few weeks later. The association of phlebitis and cruralgia should let us think of the diagnosis of vascular neoplasm. Indeed, only an early diagnosis enables a curative treatment. PMID- 11885376 TI - [Abdominal complication of hip prosthesis: peritonitis]. AB - We report a case of rare complication of hip prosthesis: peritonitis. The bad influence of radiation therapy and corticotherapy is shown. A important outcome is necessary for this patients. PMID- 11885377 TI - [Two tapes to control the main fissure of the liver]. AB - Herein we report a technique that allows a rapid and selective clamping of the left and right glissonian sheats and that secures the opening of the main fissure. The posterior face of segment IV capsula is opened immediately above the hilum on the left side of the gallbladder fossa. The tip of a right angled dissector is gently pushed in the liver substance from front to back while maintained against the hilar plate, until it arises in the caudate process just below the pedicle. A tape is used to encircle the Glisson sheath. Its inferior extremity can be picked up either on the right or the left side of the liver pedicle in order to clamp the right or the left portal pedicle, respectively. Both clamping precisely mark the anterior limit of the main fissure. Using a Kelly forceps, a second tape is introduced in the Couinaud space, between the inferior vena cava and segment one. The inferior extremity of this tape is then picked up above the Glisson sheat and allows to hang the posterior limit of the main fissure which can be securely approached. The two tapes technique cannot be applied when liver is fibrotic or when biliary ducts are dilated. PMID- 11885378 TI - ["Thyroidectomy" becoming excessive surgery?]. PMID- 11885379 TI - [Construction of a neorectum after rectal excision: colonic pouches]. AB - Rectal excision followed by low anastomosis is associated with high bowel frequency, urgency and faecal incontinence. These functional disorders results from the loss of the rectal pouch and may be also related to the damage of the anal sphincter or the loss of normal anorectal sensation. Formation of a colonic J pouch reduces the severity of the symptoms of the anterior resection syndrome mainly by decreasing bowel frequency. Creation of a J pouch may also improve the healing of coloanal anastomoses. However, there is no evidence of the role of the colonic J pouch in long term functional outcome of coloanal anastomoses. Moreover, the size of the J pouch increases with time and this may induce evacuation difficulties. Finally, the J pouch cannot be used in all patients, because of technical difficulties especially in obese men. Because the results after colonic J pouch are not perfect, new colonic pouches are developed. The caecal pouch is performed by using an ileocoecal interposition graft between the sigmoid and the anus. The transverse coloplasty is similar to that of stricturoplasty. The side-to-end coloanal anastomosis, giving a colonic blind end, is an other type of pouch. The first procedure seems technically complex with no demonstrated advantage. The second procedure is easy to construct and may be performed in all patients; however, there is a potential higher risk of leakage and functional results must be evaluated. The third procedure showed few advantages compared to a straight anastomosis. PMID- 11885380 TI - [Cephalic pancreaticoduodenectomy for adenocarcinoma of the head of the pancreas: does pylorus preservation change morbidity and prognosis?]. AB - STUDY AIM: To evaluate the influence of a pylorus-preserving on the morbidity and prognosis of patient with pancreaticoduodenectomy for adenocarcinoma of pancreas. PATIENTS AND METHODS: Between 1985 and 1999, 183 patients were operated on for pancreatic adenocarcinoma. Among them, 63 patients (40 men, mean age 63 years, range 41-77 years) had curative resection and were included in this retrospective study. They were classified according to the type of resection. In the group I, the procedure included a pylorus-preserving pancreaticoduodenectomy (n = 35). In the group II, the procedure included polar inferior gastrectomy (n = 28). The prognosis was compared. Parameters for comparison were rate of local recurrence, rate of metastatic evolution and duration of survival. RESULTS: The operative length and mortality rate (group I: 0%, group II: 3%), general (p = 0.37) and specific morbidity (p = 0.30), frequency of delayed gastric emptying were similar in the 2 groups (group I: 20%, group II: 35%, p = 0.88). The duration of naso gastic aspiration was shorter in the group I (6 days vs 8, p = 0.01). The prognosis was the same in the 2 groups (metastasis: group I: 39%, group II: 56%, p = 0.12, local recurrence: group I: 58%, group II: 43%, p = 0.09, mean survival: group I: 18 months, group II: 19 months, p = 0.77). CONCLUSION: These results suggest that pylorus preserving pancreatoduodenectomy could be performed for patients with adenocarcinoma of the head of the pancreas and does not compromise survival. PMID- 11885381 TI - [Stability of the forearm after resection of the distal ulna and proximal radius in rheumatoid arthritis: report of 11 cases]. AB - Combined resection of radial head and distal ulna could jeopardize the stability and kinematics of the forearm bones. The goals of this retrospective study was to investigate these data after resection of distal ulna and proximal radius in rheumatoid arthritis. Between 1990 and 1998, eleven patients had these bone resections combined with implantation of elbow prostheses (eight Kudo and three GSB III). Wrist surgery consisted in five wrist arthrodeses combined with Darrach procedure, four Sauve-Kapandji procedures and two isolated Darrach procedures. Mean age at surgery was 58 years and the average follow-up was 40 months. We assessed at follow-up: 1) wrist and elbow pain according to Gschwend; 2) stability of the forearm bones (cubitus valgus angle, impingement of the proximal radial stump with humerus, giving away accident of the ulnar distal stump); 3) wrist and elbow mobility. At follow-up six patients had no pain at the elbow and five had slight occasional pain. At the wrist, five patients had no pain and six slight occasional pain. Elbow motion was increased (from mean 83 degrees [50 degrees-100 degrees] to mean 110 degrees [85 degrees-135 degrees]) excepted in supination which slightly decreased (from mean 3 degrees [40 degrees-90 degrees] to mean 75 degrees [85 degrees-90 degrees]). Mean wrist mobility was impaired because of the five combined radiocarpal arthrodeses. If these five wrist arthrodeses were excluded, the mean ranges of motion were: 10 degrees in flexion, 16 degrees in extension, 2 degrees in radial deviation, 14 degrees in ulnar deviation. At follow-up, no patient had giving away accident of the ulnar distal stump nor impingement between radial stump and humerus in full flexion. Average cubitus valgus was 10 degrees. This study pointed out the predominant effect of the interosseous membrane in stability of the forearm bones. PMID- 11885382 TI - [Palliative tendon transfer for reanimation of the wrist and finger extension lag. Report of 14 transfers for radial nerve palsies and ten transfers for brachial plexus lesions]. AB - This retrospective study is based on 23 males and one female, of an average age of 36.2 years that presented to us between 1982 and 2000 with an average follow up of 61 months, with fully established paralysis of wrist and fingers extension. Fourteen patients had isolated radial nerve palsy, while ten patients had brachial plexus lesions. 1) The tendon transfer for radial nerve palsy was: PT to ECRB, FCU to ED + EPL and PL to APL + EPB; 2) for brachial plexus injury, the tendon transfer was: PT (n = 4) or FDS III or IV (n = 5) to ECRB, FCU (n = 8) or FDS IV (n = 1) to ED + EPL, PL to APL + EPB and wrist arthrodesis with transfer of FDS IV to ED + EPL and PL to APL + EPB. The results were evaluated according to the degree of wrist movement, MP extension of long fingers, opening of first commissure, thumb opposition, grip power and the subjective evaluation of results. Concerning the radial nerve palsy: results are excellent in nine cases and good in one case. An active extension of the wrist of 38 degrees was obtained as well as MP extension of 0 degree with the wrist straightened. Thumb oppositioned was conserved (Kapandji = 8.2), opening of the first commissure 40 degrees and grip power was 20 kg. Concerning the brachial plexus lesions: results are excellent in five cases and good in the other five. An active wrist extension of 32 degrees was obtained, as well as MP extension deficit of 16 degrees with wrist straightened. Opposition was concerned (Kapandji = 7.2), opening of first commissure of 38 degrees and grip power of 13 kg. The functional results are satisfactory, but the analytic study shows some effect of tenodesis of MP extension. PMID- 11885383 TI - [The VB system: a new modular osteosynthesis material involving both screws and wires]. AB - VB is an osteosynthesis system for the stabilisation of small fragments, which combines the benefits of both wires and screws. It is a modular system comprising a threaded pin and a ring. The threaded pin is first positioned. Then a ring is grasped and opened by the progressive angulation of a screwdriver. Still anchored on the screwdriver, the ring slides easily on the pin. It is clamped on the pin by simply removing the screwdriver and the pin is then cut. This modular system includes 1.8 and 1.1 mm pins and different types of rings (threaded or non threaded, with or without collars). The system is easy to handle and can be introduced using an open or percutaneous technique, allowing compression or distraction. Our preliminary series, performed in accordance with National clinical trial protocol (Huriet) consisted of 50 cases in 24 patients (five women and 19 men) with an average age of 48 years, and a follow-up of more than six months. Fourteen cases of fractures (28 implants) were treated as emergencies (two radial heads, one capitellum, one trochlea of the humerus, seven distal radius fractures, one trapezium, two metacarpals) and 12 cases (22 implants) were elective cases: arthrodesis (one trapezo-metacarpal, one intermetacarpal, two interphalangeal, two carpal), non-union (six scaphoids, one phalangeal) and one phalangeal malunion. Hardware removal was performed in 16 cases. No implant failure has been detected. One case, a DIP arthrodesis, had a suspicion of sepsis which led to the removal of the implants at six weeks. The results of this study have convinced us of the merits of the system, which combines the advantages of both wires and screws. The system allows the user to perform either distraction or compression, and to adjust the force by hand. Compared to the fixed amount of compression produced by lag screws, this feature seems to be a real step forward. PMID- 11885384 TI - Marchetti nailing with decortication and bone graft in non-unions of the two upper thirds of the humerus. AB - METHOD: Twelve patients with humeral shaft non-unions were treated using a Marchetti-Vicenzi nailing. The fractures site was decorticated and bone graft added. RESULTS: Fracture healing was obtained in all cases. The mean healing time was 4.7 months. The range of motion of the shoulder was excellent in nine patients, moderate in two and poor in one. The elbow had an excellent range of motion in ten patients, moderate in one and poor in one. The functional result was excellent in nine patients, good in two, and fair in one. CONCLUSION: Marchetti-Vicenzi nailing with bone grafting appears to be a good method for the treatment of humeral shaft non-unions. It is technically easy and its results are satisfactory. PMID- 11885385 TI - [Juvenile aponeurotic fibroma. A case report with a review of the literature]. AB - Juvenile aponeurotic fibroma is a rare benign tumour which occurs mainly in young patients under 20 years of age and especially during childhood. Clinical presentation is a unique, hard and painless tumour of the palm or sole. The treatment commonly accepted for this locally recurrent tumour is complete excision with function preservation. In this paper, we report on a case of juvenile aponeurotic fibroma of the thenar area with a thickening of the first metacarpal bone shaft. Bone involvement in juvenile aponeurotic fibroma is a very rare condition. We only found one report of a comparable case in literature. PMID- 11885386 TI - [Aseptic bone necrosis following reimplantation of a degloving finger]. AB - We report a case of microsurgical replantation of a degloved finger in a manual worker. Four months following replantation, avascular necrosis of the middle and distal phalanges was apparent. Amputation at the level of the proximal phalanx was performed. Re-plantation is the solution of choice for such degloving injuries, but a different flap can be used if replantation is not possible. Avascular necrosis of bone is an unfrequent complication, but surgeons should be aware of it. PMID- 11885388 TI - [Island bone graft from the second metacarpal to the first metacarpal]. AB - We report a case of reconstruction of the first metacarpal by using an extended vascularized bone graft taken from the distal part of the second metacarpal. Following a gunshot injury, a 57 year-old man presented with a thumb amputated at the interphalangeal joint level and an index finger amputated at the metacarpophalangeal joint level. After lengthening of the first metacarpal over a six weeks period, the bone gap was filled using a vascularized 2.4 cm-length piece of the second metacarpal. This graft was vascularized by the dorsal vascular network of the first web space. At two months, the bone fixation was removed. At five months, the bony integration was complete. PMID- 11885387 TI - Hamatometacarpal fracture-dislocation: distinctive three dimensional computed tomographic appearance. AB - We report here two fully documented cases of hamatometacarpal fracture dislocation following trauma and treated in our hospital. In our cases, the patients suffered hamate fracture in association with metacarpal dislocation. In the first case, a dorsal oblique fracture of the hamate was associated with a dorsal dislocation of the base of the fourth metacarpal. In the second case, a dorsal oblique fracture of the hamate was not associated with a dorsal dislocation of the base of the fifth metacarpal. This diagnosis should be suspected on initial review of plain radiographs, which must include an oblique view because of diagnostic difficulty for this injury. We recommend three dimensional computed tomography (3D-CT) in any patient presenting with pain after blunt trauma to the hand to prevent in diagnosis. Open reduction and internal fixation of the fracture is indicated and relevant for displaced fracture. PMID- 11885389 TI - [Four bone versus capito-lunate limited carpal fusion. Report of 40 cases]. AB - Fourty patients with limited carpal fusion have been retrospectively reviewed. The aim of this study was to compare the results of four bone fusions (30 wrists) versus capitolunate fusion (11 wrists). Follow-up averaged 30 months with a range of 15-96 months. Twelve patients presented SLAC-wrist (scapho-lunate advanced collapse) and fourteen with SNAC-wrist (sapho-non union advanced collapse). There were seven cases of primitive wrist arthritis, one mid-carpal instability, one sequella of Fenton's syndrome, one Preiser's disease and two Kienbock's disease. All 40 patients were evaluated by the same observer. In terms of range of motion, the capito-lunate fusion led to better results than the four bone fusions with a gain of 10 degrees in volar flexion and 12 degrees in radial deviation. Results in term of post operative pain are similar in the two groups of patients with 90% painless wrist in four bone fusion and 81% with capito-lunate fusion. Results for strength were equivalent. Radiological bone fusion was obtained within nine weeks. Absence of fusion was observed in two patients with capito-lunate fusion. Our result in terms of joint motion and strength are similar to those found in the literature. Correction or not of the DISI deformity during the procedure did not affect the results (on 19 patients). The Four bone fusion procedure is still a good treatment in SLAC or SNAC wrist. Capito-lunate fusion remains a good choice, despite the risk of non-fusion. PMID- 11885390 TI - [Divergent dislocation of the elbow in an adolescent. A case report]. AB - A case of simultaneous proximal radio ulnar joint divergent dislocation combined with a bone avulsion of the coronoid apophysis of the same elbow in a 16 years old girl is presented. After a closed reduction and two weeks of plaster immobilization, normal function of the elbow was recovered within three months. There are only ten cases reported of such a divergent elbow dislocation in modern literature. PMID- 11885391 TI - [Reliability of global interdigital pinch (vice pinch) dynamometral values measured by Microfet 2]. AB - OBJECTIVE: To study test-retest reliability of pinch grip strength with a two month gap in three upper limb positions. To study factors causing variability of peak force. MATERIAL AND METHODS: Forty healthy control volunteers, sex-ratio 1, mean age 36.6 years (sd = 9.9), were separated into four groups of ten persons. The Microfet 2 dynamometer was used to measure peak force developed during pinch grip. The measurements were made in three positions: upper limb parallel to body, arm flexed at 90 degrees and during the Roos manoeuvre. Test-retest reliability was evaluated after two months. Statistical analysis used the correlation test (p = 0.05). RESULTS: Reliability proved to be excellent in all three positions and in all four groups. Dynamometer values were stable between the beginning and end of the Roos manoeuvre in this healthy population. There were no significant differences between peak force measured in the three positions and between left and right upper limbs. DISCUSSION: The Roos manoeuvre gives a stable and reliable result in a population without thoracic outlet syndrome. Dynamometer tests are also reliable in the usual upper limb positions, in accordance with other studies. CONCLUSION: This dynamometric test should be useful in assessing the severity of thoracic outlet syndrome and in demonstrating the therapeutic efficacy of different treatments. PMID- 11885392 TI - CDC updates childhood immunization schedule. PMID- 11885393 TI - Recap of FDA product approvals--2001. PMID- 11885394 TI - FDA offers guidance on prophylaxis for exposure to radioiodines. PMID- 11885395 TI - Alitretinoin. PMID- 11885396 TI - Formoterol fumarate. PMID- 11885397 TI - Update on natural product--drug interactions. AB - The interactions of natural products with drugs are discussed. Interactions between natural products and drugs are based on the same pharmacokinetic and pharmacodynamic principles as drug-drug interactions. Clinically important interactions appear to involve effects on drug metabolism via cytochrome P-450 isoenzymes, impairment of hepatic or renal function, and other possible mechanisms. Natural products that have been reported to interact with drugs in humans include coenzyme Q10, dong quai, ephedra, Ginkgo biloba, ginseng, glucosamine sulfate, ipriflavone, melatonin, and St. John's wort. In many cases, more research is needed to confirm these interactions and to determine whether other natural products may also interact with drugs. To effectively counsel patients about interactions involving natural products, pharmacists should be familiar with the most commonly used products and have access to information on more obscure products. In view of the less than stringent provisions of the Dietary Supplement Health and Education Act, pharmacists should consult reliable, independent sources of information on natural products rather than rely on literature provided by manufacturers. Pharmacists should recommend only those products that are manufactured to high quality-control standards. Natural products can interact with drugs and with other natural products by the same mechanisms as drugs. PMID- 11885398 TI - Use of patients' own medications in small hospitals. AB - The prevalence, management, and adverse events associated with the use of a patient's personal medications in hospitals were studied. A questionnaire comprised of 17 questions was mailed in November 1999 to the pharmacy directors of a random sample of 300 small (< or = 200-bed capacity) hospitals selected from the American Hospital Association 1999 membership directory. A follow-up mailing was sent to nonrespondents in early December 1999. The total usable response rate was 54.6%. The mean bed capacity was 76.6, and 70.8% of facilities had < or = 100 beds. Most facilities provided acute care, were nonprofit organizations, and were located in rural areas. A majority (90%) of the pharmacy directors surveyed allowed patients to use their own medications in the hospital. Elderly patients were most likely to bring their personal medications to use in the hospital, and pharmacists were the health professionals most likely to identify patients' personal medications. Circumstances in which patients were allowed to use their own medication, provided there was a physician's order, included prepackaged courses of therapy or antimicrobial courses and nonformulary medications, excluding controlled substances. Loss of personal medication and medication errors were the most frequently identified problems with allowing patients to use personal medications. Most small hospitals allowed the use of patients' personal medications; however, there was a wide variation in the circumstances for which the use of these medications was allowed. PMID- 11885399 TI - Effectiveness of preprinted order forms in promoting perioperative beta-blocker use. AB - The Notes section welcomes the following types of contributions: (1) practical innovations or solutions to everyday practice problems, (2) substantial updates or elaborations on work previously published by the same authors, (3) important confirmations of research findings previously published by others, and (4) short research reports, including practice surveys, of modest scope or interest. Notes should be submitted with AJHP's manuscript checklist. The text should be concise, and the number of references, tables, and figures should be limited. PMID- 11885400 TI - Stability of oral suspensions of ursodiol made from tablets. PMID- 11885401 TI - Multidisciplinary approach to improving treatment of community-acquired pneumonia. PMID- 11885402 TI - Ideal principles and characteristics of a fail-safe medication-use system. PMID- 11885403 TI - Efficacy of HMG-CoA reductase inhibitors in treating osteoporosis. AB - The Therapy Consultation section provides brief advice, in question-and-answer format, on how to handle specific drug-therapy problems. Readers are invited to submit questions, which will be referred to a consultant for an answer. The problems should not be highly specialized or unusual, nor should they be mundane. Questions are answered briefly, largely on the basis of judgment and personal experience of the consultant, although selected references may be cited to substantiate the consultant's advice. PMID- 11885404 TI - Drug administration through enteral feeding catheters. PMID- 11885405 TI - Herbal product use among anticoagulation clinic patients. PMID- 11885406 TI - Goals of statin conversion program. PMID- 11885407 TI - Value of residency projects. PMID- 11885408 TI - Optimal antimicrobial therapy for sepsis. AB - The selection of appropriate antimicrobial therapy for patients with sepsis is discussed. Antimicrobial selection is based on the most likely source of infection, the most common pathogens at that site, knowledge of antimicrobial susceptibility patterns in the local community and institution, and host factors. Prompt initiation of appropriate antimicrobial therapy is essential for achieving the best possible outcomes. Defining optimal antimicrobial therapy for sepsis is difficult because few clinical studies have specifically addressed this. Sepsis has traditionally been associated with gram-negative infections, but studies indicate that gram-positive pathogens commonly cause sepsis and are associated with high mortality. Enterobacteriaceae, species, Pseudomonas aeruginosa, and Staphylococcus aureus are the most common pathogens in sepsis patients and should be considered in the selection of empirical therapy. Antimicrobial therapy should be initiated as soon as samples have been obtained for culture; empirical therapy should cover a very broad spectrum of organisms, with consideration given to the most likely pathogens for the infection site. An antipseudomonal beta-lactam with or without an aminoglycoside is recommended for initial therapy in most patients. The role of monotherapy versus combination therapy is controversial. Routine use of vancomycin and antifungal therapy as part of initial regimens should be discouraged but may be justified in specific circumstances. PMID- 11885409 TI - Recombinant human activated protein C in severe sepsis. AB - The role of activated protein C (APC) in coagulation, inflammation, and fibrinolysis and the pharmacology, pharmacokinetics, and trials of recombinant human activated protein C (rhAPC), or drotrecogin alfa (activated), in sepsis are described. Protein C, a naturally occurring vitamin K-dependent serine protease in the blood, remains inactive until exposed to the thrombin-thrombomodulin complex. This change between the inactive and active forms occurs constantly in humans and serves to balance the coagulation cascade. APC functions in concert with protein S as an anticoagulant, a fibrinolytic agent, and an antiinflammatory agent. In response to serious infection, a procoagulant process is activated leading to thrombin and fibrin deposition in small vessels that results in decreased blood flow, decreased oxygen delivery, and organ failure. The body's natural defense during severe sepsis is to activate protein C through the thrombin-thrombomodulin complex in an attempt to restore the imbalance of the hemostatic systems. However, APC has a short half-life, and the pool of circulating protein C is rapidly depleted in severe sepsis. Low protein C levels have been correlated with poor outcome in patients with severe sepsis and in animal models. These observations led to a Phase III safety and efficacy trial of drotrecogin alfa (activated) that demonstrated a significant improvement in mortality compared with placebo (24.7% versus 30.8%). This 6.1% absolute difference in mortality translates to a 19.4% reduction in relative risk of death in the treated patients. The proper use of drotrecogin alfa (activated) will require careful consideration of appropriate patients to treat and further studies in patient populations that were excluded from the Phase III trial, as well as possible modification of dosing schemes on the basis of patient response. PMID- 11885410 TI - Innovation in sepsis management. Introduction. PMID- 11885411 TI - Management challenge with drotrecogin alfa (activated). AB - Clinical trial results for drotrecogin alfa (activated) are discussed in terms of potential effect on morbidity and total cost of care. Interdisciplinary collaboration is crucial for improving care in health systems; teamwork has been shown to improve outcomes in intensive care and reduce costs. The treatment of sepsis is costly and resource intensive. With existing treatments, drugs account for a lower percentage of the total cost than does care in the intensive care unit. Study results indicate that drotrecogin alfa (activated) reduces mortality in severe sepsis without a major morbidity penalty in 28 days. No difference was found in total resource use over 28 days between patients who received the drug and those who received placebo, even though more patients who received drotrecogin alfa (activated) survived and thus required treatment. Estimated costs per life-year saved fall within the cost-effective range. When drotrecogin alfa (activated) becomes available for clinical use, pharmacists must systematically assess this new therapeutic tool in terms of health value. PMID- 11885412 TI - Pathophysiology of sepsis. AB - The roles of inflammation and coagulation in the pathophysiology of sepsis are described. Sepsis results when an infectious insult triggers a localized inflammatory reaction that then spills over to cause systemic symptoms of fever or hypothermia, tachycardia, tachypnea, and either leukocytosis or leukopenia. These clinical symptoms are called the systemic inflammatory response syndrome. Severe sepsis is defined by dysfunction of one of the major organ systems or unexplained metabolic acidosis. The inflammatory reaction is mediated by the release of cytokines, including tumor necrosis factor-alpha, interleukins, and prostaglandins, from neutrophils and macrophages. The cytokines activate the extrinsic coagulation cascade and inhibit fibrinolysis. These overlapping processes result in microvascular thrombosis; thrombosis is one potential factor producing organ dysfunction. Activation of the coagulation system leads to consumption of endogenous anticoagulants (e.g., protein C and antithrombin); this may be an important factor in the development of microvascular coagulation. Antiinflammatory mediators as well as inflammatory mediators have a role in sepsis, and an excess of either can result in poor patient outcomes. Sepsis is a complex syndrome involving activation of a variety of systems. PMID- 11885413 TI - Current strategies for managing the patient with sepsis. AB - Key elements of the current approach to treating sepsis are reviewed, and examples are given to illustrate the difficulty of designing and evaluating trials in sepsis. A patient with sepsis is likely to have symptoms characteristic of the systemic inflammatory response syndrome. Initially, ruling out noninfective causes, locating the site of infection, and obtaining cultures before beginning antimicrobial therapy are critical. Aggressive fluid resuscitation and hemodynamic support are used to restore tissue perfusion and normalize cellular metabolism. Vasopressor therapy with dopamine or norepinephrine is needed in patients unresponsive to fluid resuscitation. Dobutamine should be administered in patients whose cardiac output is inadequate despite optimization of fluids and pressors. Supportive care includes deep vein thrombosis prophylaxis, nutrition support, stress ulcer prophylaxis, and management of acute lung injury. Attempts to modify the sepsis response and improve the outcome in these patients have yielded limited benefits. Recent small studies have shown benefits with low-dose hydrocortisone in patients with refractory sepsis. One challenge in study design is that a therapy may target a subset of patients that cannot be identified at the outset. Management of patients with suspected or documented sepsis focuses on hemodynamic support, appropriate antimicrobial therapy, and other supportive care. PMID- 11885414 TI - Prospects for the phytoremediation of organic pollutants in Europe. PMID- 11885415 TI - Exploiting plant metabolism for the phytoremediation of persistent herbicides. AB - Weed control by herbicides has helped us to create the green revolution and to provide food for at least the majority of human beings living today. However, some herbicides remain in the environment and pose an ecological problem. The present review describes the properties and fate of four representative herbicides known to be presistent in ecosystems. Metabolic networks are depicted and it is concluded that removal of these compounds by the ecologically friendly technique of phytoremediation is possible. The largest problem is seen in the uptake of the compounds into suitable plants and the time needed for such an approach. PMID- 11885416 TI - Phytoremediation of polyaromatic hydrocarbons, anilines and phenols. AB - Phytoremediation technologies based on the combined action of plants and the microbial communities that they support within the rhizosphere hold promise in the remediation of land and waterways contaminated with hydrocarbons but they have not yet been adopted in large-scale remediation strategies. In this review plant and microbial degradative capacities, viewed as a continuum, have been dissected in order to identify where bottle-necks and limitations exist. Phenols, anilines and polyaromatic hydrocarbons (PAHs) were selected as the target classes of molecule for consideration, in part because of their common patterns of distribution, but also because of the urgent need to develop techniques to overcome their toxicity to human health. Depending on the chemical and physical properties of the pollutant, the emerging picture suggests that plants will draw pollutants including PAHs into the plant rhizosphere to varying extents via the transpiration stream. Mycorrhizosphere-bacteria and -fungi may play a crucial role in establishing plants in degraded ecosystems. Within the rhizosphere, microbial degradative activities prevail in order to extract energy and carbon skeletons from the pollutants for microbial cell growth. There has been little systematic analysis of the changing dynamics of pollutant degradation within the rhizosphere; however, the importance of plants in supplying oxygen and nutrients to the rhizosphere via fine roots, and of the beneficial effect of microorganisms on plant root growth is stressed. In addition to their role in supporting rhizospheric degradative activities, plants may possess a limited capacity to transport some of the more mobile pollutants into roots and shoots via fine roots. In those situations where uptake does occur (i.e. only limited microbial activity in the rhizosphere) there is good evidence that the pollutant may be metabolised. However, plant uptake is frequently associated with the inhibition of plant growth and an increasing tendency to oxidant stress. Pollutant tolerance seems to correlate with the ability to deposit large quantities of pollutant metabolites in the 'bound' residue fraction of plant cell walls compared to the vacuole. In this regard, particular attention is paid to the activities of peroxidases, laccases, cytochromes P450, glucosyltransferases and ABC transporters. However, despite the seemingly large diversity of these proteins, direct proof of their participation in the metabolism of industrial aromatic pollutants is surprisingly scarce and little is known about their control in the overall metabolic scheme. Little is known about the bioavailability of bound metabolites; however, there may be a need to prevent their movement into wildlife food chains. In this regard, the application to harvested plants of composting techniques based on the degradative capacity of white-rot fungi merits attention. PMID- 11885417 TI - Prospects and limitations of phytoremediation for the removal of persistent pesticides in the environment. AB - The environmental problems that have arisen from the use of persistent pesticides in the past, and potential sources of further contamination have been discussed. The potential and limitations of phytoremediation for removal of pesticides in the environment have been reviewed. The enzymatic processes in plants that are known to be involved in phytodegradation of pesticides, and possibilities for enhancing them have also been discussed. PMID- 11885418 TI - Biological remediation of explosives and related nitroaromatic compounds. AB - Nitroaromatics form an important group of recalcitrant xenobiotics. Only few aromatic compounds, bearing one nitro group as a substituent of the aromatic ring, are produced as secondary metabolites by microorganisms. The majority of nitroaromatic compounds in the biosphere are industrial chemicals such as explosives, dyes, polyurethane foams, herbicides, insecticides and solvents. These compounds are generally recalcitrant to biological treatment and remain in the biosphere, where they constitute a source of pollution due to both toxic and mutagenic effects on humans, fish, algae and microorganisms. However, relatively few microorganisms have been described as being able to use nitroaromatic compounds as nitrogen and/or carbon and energy source. The best-known nitroaromatic compound is the explosive TNT (2,4,6-trinitrotoluene). This article reviews the bioremediation strategies for TNT-contaminated soil and water. It comes to the following conclusion: The optimal remediation strategy for nitroaromatic compounds depends on many site-specific factors. Composting and the use of reactor systems lend themselves to treating soils contaminated with high levels of explosives (e.g. at former ammunition production facilities, where areas with a high contamination level are common). Compared to composting systems, bioreactors have the major advantage of a short treatment time, but the disadvantage of being more labour intensive and more expensive. Studies indicate that biological treatment systems, which are based on the activity of the fungus Phanerochaete chrysosporium or on Pseudomonas sp. ST53, might be used as effective methods for the remediation of highly contaminated soil and water. Phytoremediation, although not widely used now, has the potential to become an important strategy for the remediation of soil and water contaminated with explosives. It is best suited where contaminant levels are low (e.g. at military sites where pollution is rather diffuse) and where larger contaminated surfaces or volumes have to be treated. In addition, phytoremediation can be used as a polishing method after other remediation treatments, such as composting or bioslurry, have taken place. This in-situ treatment method has the advantage of lower treatment costs, but has the disadvantage of a considerable longer treatment time. In order to improve the cost-efficiency, phytoremediation of nitroaromatics (and other organic xenobiotics) could be combined with bio-energy production. This requires, however, detailed knowledge on the fate of the contaminants in the plants as well as the development of efficient treatment methods for the contaminated biomass that minimise the spreading of the contaminants into the environment during post harvest treatment. PMID- 11885419 TI - Sulphonated aromatic pollutants. Limits of microbial degradability and potential of phytoremediation. AB - Many synthetic sulphonated aromatic compounds are used as starting material to produce dyes and pigments, or are released as by-products in the effluents of the textile and dye industry. A large number of these chemicals are poorly biodegradable and cannot be eliminated by classical wastewater treatment plants. To limit the impact of these pollutants on the environment, new processes, based on the use of higher plants (constructed wetlands or hydroponic systems), are under development. Detergents and surfactants are essential for both industrial and domestic applications, the most important family being the alkylbenzene sulphonates. Originally, the alkyl side chains were branched and thus recalcitrant to biodegradation. Therefore, they have been replaced by linear alkylbenzene sulphonates. Although more acceptable, present formulations still have adverse environmental and toxic effects. In this context, phytoremediation appears to be a promising approach to remove these compounds from contaminated soils and waters. PMID- 11885420 TI - Phytoremediation to increase the degradation of PCBs and PCDD/Fs. Potential and limitations. AB - Phytoremediation is already regarded as an efficient technique to remove or degrade various pollutants in soils, water and sediments. However, hydrophobic organic molecules such as PAHs, PCBs and PCDD/Fs are much less responsive to bioremediation strategies than, for example, BTEX or LAS. PCDD/Fs and PCBs represent 3 prominent groups of persistent organic pollutants that share common chemical, toxicological and environmental properties. Their widespread presence in the environment may be explained by their chemical and biological stability. This review considers their fate and dissipation mechanisms. It is then possible to identify major sinks and to understand biological activities useful for remediation. Public health and economic priorities lead to the conclusion that alternative techniques to physical treatments are required. This review focuses on particular problems encountered in biodegradation and bioavailability of PCDD/Fs and PCBs. It highlights the potential and limitations of plants and micro organisms as bioremediation agents and summarises how plants can be used to augment bacterial activity. Phytoremediation is shown to provide some new possibilities in reducing risks associated with dioxins and PCBs. PMID- 11885422 TI - Assistance for a dental student. PMID- 11885421 TI - Remediation of BTEX and trichloroethene. Current knowledge with special emphasis on phytoremediation. AB - The widespread use of industrial chemicals in our highly industrialized society has often caused contamination of large terrestrial and marine areas due to the deliberate and accidental release of organic pollutants into the soil and groundwater. In this review, environmental problems arising from the use of chlorinated solvents and BTEX compounds are described, and an overview about active management strategies for remediation with special emphasis on phytoremediation are presented to achieve a reduction of the total mass of chlorinated solvents and BTEX compounds in contaminated areas. Phytoremediation has been proposed as an efficient, low-cost remediation technique to restore areas contaminated with chlorinated solvents and BTEX compounds. The feasibility of phytoremediation as a remediation tool for these compounds is discussed with particular reference to the uptake and metabolism of these compounds, and a future perspective on the use of phytoremediation for the removal of chlorinated solvents and BTEX compounds is given. PMID- 11885423 TI - Effectiveness of three root canal medicaments to eliminate Actinomyces israelii from infected dentinal tubules in vitro. AB - The persistence of anaerobic bacteria in the root canal system often leads to treatment failure. One possible reason for this may be the retention of micro organisms in the dentinal tubules of root canal walls. This study was performed to compare the effectiveness of two root canal medicaments and a chlorhexidine solution in disinfecting Actinomyces israelii-infected root canal walls and dentinal tubules in vitro. Dentinal tubules of root canal walls of human teeth were experimentally infected with A. israelii. The root canals were exposed to either iodine potassium iodide, calcium hydroxide or 2% chlorhexidine for periods of 3, 7 and 60 days. At the end of the medication periods samples were removed at different depths and tested for A. israelii viability. Chlorhexidine was the only disinfectant that was able to eliminate A. israelii from all the samples after 3, 7 as well as 60 days while 25% of the specimens treated with iodine potassium iodide and 50% of the specimens treated with calcium hydroxide still had viable A. israelii after treatment. It is clear from this study that 2% chlorhexidine is superior to iodine potassium iodide and calcium hydroxide in its ability to remove A. israelii from infected dentinal tubules. However, in vivo trials need to be undertaken before its clinical use can be recommended. PMID- 11885424 TI - Does the lead apron and collar always reduce radiation dose? AB - The possibility that personal lead shielding devices can increase absorption of radiation has not been entertained. The purpose of the present investigation specifically was to determine whether pituitary dose might be increased when a leaded apron and thyroid collar are used. Thermoluminescent dosimeters (TLDs) were used to measure absorbed dose. They were calibrated at the kVp used in the clinical situation and a calibration curve relating light output to dose was generated. Lithium fluoride TLD discs were placed in the pituitary gland region of a Rando-Alderson female human phantom. The equivalent of 100 transpharyngeal exposures were delivered. The resultant light output from recovered dosimeters was converted to a uGy value using the calibration curve. The experiment was repeated using a 0.25 mm lead equivalent collar and apron fitted to the phantom in the customary manner. The entire process was repeated in order to have 30 dosimeters for the unshielded and 30 dosimeters for the shielded conditions. A further 30 dosimeters were sham irradiated and served as controls. A statistical comparison between unshielded and shielded conditions was performed. When the leaded apron and thyroid collar were used the absorbed dose to the pituitary gland was increased significantly (P < 0.05). Following this a second group, using a different dosimetry system and a male phantom repeated the experiment. In both cases, the shielded phantom received significantly higher dose to the pituitary region than the unshielded. PMID- 11885425 TI - Perceptions of oral health: the South African Demographic and Health Survey of 1998. AB - Oral diseases are widespread in South Africa and affect large numbers of people in terms of pain, tooth loss, disfigurement, loss of function and even death. The majority of South Africans have no access to private services and are dependent on the government for oral health care services, but less than 10% of the population utilises public oral health services. This underutilisation is due to limited resources and inaccessibility. This article reports on the data collected on adults, 15 years and older, from the South African Demographic and Health Survey carried out in 1998, relating to perceptions of oral health. The questions dealt with oral health problems, utilisation of oral health services, loss of natural teeth, oral health practices and knowledge of water fluoridation. The results are discussed in terms of age, gender, education, place of residence, province and classification according to the previous population registration act. A high proportion (36%) of people had experienced oral health problems. Teeth problems were most commonly reported in the higher age groups, non-urban areas, Eastern Cape, Northern Cape and Free State, people with little education and those classified as non-urban Africans. Significant differences were found in regard to periodontal disease, tooth loss, knowledge of fluoride between groups according to age, geographic location, race and level of education. 62% of the respondents reported that they had lost some of their natural teeth and in some communities almost a third of the respondents were edentulous. In comparing the goals of the current draft national oral health policy with the findings of this survey the following implications for policy development should be noted: A higher priority needs to be given to oral health issues; Improve access by increasing primary health care facilities through the delivery of oral health care services; The high prevalence of hepatitis and HIV/AIDS infection poses a higher risk to oral health personnel and the public and The successful implementation of water fluoridation depends upon public knowledge, understanding and support. PMID- 11885426 TI - Bacterial contamination of dental handpieces. AB - Bacterial contamination of water- and air-lines was investigated after artificial contamination and clinical use of dental handpieces. The effect of lubrication, disinfection and autoclaving on this contamination was also addressed. Bacterial growth was recorded in both the air- and water-lines during clinical and artificial contamination, with water-lines showing heavier contamination than air lines. External and internal surfaces of handpieces still yielded bacterial growth after lubrication and disinfection, while heat sterilisation (autoclaving) rendered both internal and external surfaces sterile. PMID- 11885427 TI - [Polymerization shrinkage by 4 different types of dental materials]. AB - Forces developing during polymerisation of dental resins cause tension in the material to increase, with possible subsequent distortion of the bond. This study was undertaken to determine and compare polymerisation shrinkage in four different light-cured dental resins. A modified dilatometer was used to determine volumetric changes during polymerisation in 60 seconds of Z250 and Filtec flow from 3 M, and DyractAP and Dyract from Dentsply. Statistical analysis revealed that Dyract Flow shrinks significantly more (P < 0.05) during polymerisation in the first, as well as the last 10 seconds when compared with the other three materials and Z250 significantly less. An increase in the amount of fillers in the composition of the material leads to a decrease in polymerisation shrinkage, while an increase in the monomer concentration gives rise to more shrinkage. Stress, that builds up during the polymerisation process, is reduced by the elasticity of the material. The shrinkage of the flowables is therefore counteracted by their reduced rigidity. PMID- 11885428 TI - Human vaginal epithelium and the epithelial lining of a cyst model constructed from it: a comparative light microscopic and electron microscopic study. AB - The light microscopic features and keratin filament distribution of human vaginal epithelium resemble those of buccal mucosa. We used vaginal epithelium to establish a human cyst model in immunodeficient mice. To strengthen the view that this experimental cyst is a suitable model to study mucosal diseases, we compared specific light microscopic and ultra-structural features of vaginal epithelium and the epithelial lining of the cyst. Nineteen cyst walls and 6 specimens of vaginal mucosa, which had been used to establish the cysts, were examined. We counted the number of cell layers of 17 cyst linings and the 6 vaginal specimens. Surface keratinisation was evaluated on sections stained with the Picro-Mallory method. To demonstrate intercellular lamellae and membrane coating granules 2 cyst linings were examined ultra-structurally. The epithelium lining of the cyst wall was thinner than that of vaginal mucosa but the surface keratinisation and ultra-structural features of the intercellular lamellae and membrane coating granules were similar. We concluded that vaginal mucosa is a useful substitute for oral mucosa in the cyst model. PMID- 11885429 TI - Prevalence and impact of dental pain in 8-10-year-olds in the western Cape. AB - The purpose of this study was to investigate the prevalence of acute dental pain in 8-10-year-old schoolchildren in the Western Cape. The study sample was drawn from schools in Stellenbosch, Malmesbury, Mitchells Plain, Mossel Bay and Robertson. Eighty eight per cent reported that they had experienced dental pain and 70% within the last two months. The prevalence and severity of pain in this study is much higher than reported studies in England and the USA. A high percentage of children not only live with pain on a daily basis, but also missed school on account of it (70%). There is an urgent necessity for oral health to be given a greater priority within the district health system to improve access to integrated oral health care services with appropriate preventive and treatment measures. PMID- 11885430 TI - Extranodal non-Hodgkin's lymphoma of the maxillofacial region: analysis of 88 consecutive cases. AB - Examination of the records of 88 consecutive patients with extranodal maxillofacial non-Hodgkin's lymphoma (ENHL) was undertaken. Each patient's complete record was reviewed. Males outnumbered females by 1.7:1. Age at diagnosis ranged from 22 to 94 years (median 60.0). Affected anatomic sites included: maxillary sinus (22), nasal cavity (8), maxilla (13), mandible (8), salivary glands (14), and other (23). The most common presenting symptom was a non-painful mass. Associated dental symptoms were present in 72 patients and included intraoral swelling, pain, and loose teeth. Treatment included chemotherapy and radiation with a follow-up of 1-25 years. Treatment trends indicate a shift towards multimodal therapy. Non-Hodgkin's lymphoma of the head and neck, if discovered early, has an excellent prognosis. PMID- 11885431 TI - Perceptions of fluorosis in northern Cape communities. AB - The objective of the study was to determine the perception of fluorosis in communities living in the Northern Cape Province of South Africa where there is a considerable range in fluoride levels of drinking water. The fluoride levels of the drinking water were categorised as suboptimal (0.40-0.60 ppmF), optimal (0.99 1.10 ppmF) or supra-optimal (1.70-2.70 ppmF). The teeth of 694 children aged 6, 12 and 15 years were examined. Dental fluorosis occurred among children of all ages in all areas studied. As anticipated there appears to be a direct relationship between fluoride levels in the drinking water and levels of dental fluorosis, and the severity of the condition increased with an increase in levels of fluoride in the water supplies. Children in low fluoride areas showed some form of mild fluorosis (37% very mild and 17% mild). However, 19% of this group experienced moderate or severe forms of fluorosis. In areas with optimal levels of fluoride 30% of children showed a questionable form of fluorosis and 21% mild fluorosis. Moderate or severe forms of fluorosis were recorded in 31% of children in the optimal fluoride area. The Community Fluorosis Index (CFI) scores for the sub-optimal and optimal areas were of medium public health significance and for the supra-optimal area of very high public health significance. Of concern is the high percentage of children (45%) in the supra-optimal area with severe forms of fluorosis. The awareness and concern for stains on teeth were mostly expressed by children with moderate or severe fluorosis. This study suggests that the proposed fluoride concentration (not more than 0.7 ppmF) prescribed in the Regulations on Fluoridating the Water Supplies for South Africa would minimise the risk of dental fluorosis. PMID- 11885432 TI - The dental health of 12-year-old children whose diets include canned fruit from local factories: an added risk for caries? AB - OBJECTIVE: To determine the dental health of 12-year-old children from households that are supplied with sweetened canned fruit from local factories. METHODS: The DMFS and DMFT indices were determined for 12-year-old pupils from 6 schools, 3 of which are situated in communities with fruit canning factories. Subjects were questioned about the workplace of household members and supplies of sweetened canned fruit from factories. To compare indices we used a two-way analysis of variance, for multiple comparisons the Bonferroni Test and for proportions the Chi-Squared Test. Fluoride content of the drinking water was measured for each school. RESULTS: The fluoride content for all schools was less than 0.1 ppm. In only 2 schools were the majority of pupils from households that were supplied regularly with canned fruit from factories. An analysis of the DMFT data of the children in all the schools showed that there were significantly fewer children with a DMFT = 0 and significantly more with a DMFT = 4+ in the 2 schools. The mean DMFS and DMFT of children in households supplied with canned fruit were significantly higher than those children without the supply. CONCLUSION: The supply of sweetened canned fruit to households may be an added risk to dental health for the children in that household. PMID- 11885433 TI - The use of subepithelial connective tissue grafts in the treatment of marginal recession defects--a surgical approach. PMID- 11885434 TI - [Advancements at the Faculty of Dentistry, now the School for Oral Health Science -an overview]. AB - The aim of this communication is to give an overview of the contributions made by the faculty of Dentistry, now School of Oral Health Science, at scientific meetings of the SA Division of the IADR since 1970, the year that research started in the faculty. With the increase of staff from 1970 research started in the faculty. With the increase of staff from 1970 research papers increased in leaps and bounds. In 1970 one paper was presented in 1978 there were 15 and in 1980, 27. The abstracts of 423 presentations have been published in the Journal of Dental Research - 17% of all the abstracts of all the papers read at IADR meetings. The majority of papers from this faculty concerned pathological conditions of the mouth (77 or 18.2%), followed by community-oriented research (8.3%), fluoride (8%), and microbiology and orthodontics (7.8% respectively). Also prominent were papers on rontgenology, oral biology, pharmacology, periodontics, prosthetics, forensic dentistry, oral surgery and research on lead uptake. Epidemiology featured in 19.9% of the research papers. Of note are the number of doctorates awarded: 20 PhDs, 4 senior doctorates (DSc), 1 DEd, DSc from the University of Pretoria and an Honorary Doctorate also from Pretoria. PMID- 11885435 TI - Non-Hodgkin's lymphomas of Waldeyer's ring: a clinicopathological and immunological study of 64 cases in the western Cape. AB - The clinicopathological and immunological features of 64 cases of primary extranodal non-Hodgkin's lymphoma (NHLs) that occurred in Waldeyer's ring (WR) were examined. The objective was to compare the findings of this study with those of previous studies. The age at presentation, sex ratio, and site of occurrence of these tumours within WR concurred with that of other studies. Diffuse large cell lymphomas were the most prevalent in this study. Most T-cell NHLs occurred in the nasopharynx where they constituted 28% of all NHLs in that site. This indicates a higher incidence of nasopharyngeal T-cell NHLs in South Africa as compared with other Western countries. PMID- 11885436 TI - The role of the dentist in detection of carotid atherosclerosis. AB - Cerebrovascular accidents (CVA), or stroke, afflict 731,000 Americans each year, with 165,000 of these individuals dying. Stroke is a major cause of death and disability throughout the world, including southern Africa. Atherosclerosis related formation of thrombi and emboli at the bifurcation of the common carotid artery and proximal internal carotid artery represents a common cause of stroke. The detection of carotid atherosclerosis by dentists using panoramic radiographs recently has been presented to the public through television news stories and the press, but many dentists still do not know how to interpret panoramic radiographs for detection of this condition. This communication illustrates examples in which carotid atherosclerosis was detected using panoramic radiography. Differential diagnoses are presented. Since not every carotid plaque calcifies, panoramic radiography should never be used alone to exclude the possibility of carotid atherosclerosis. It should also be remembered that the mere presence of calcified carotid plaque is not necessarily a reflection on the degree of carotid stenosis. Definitive diagnosis and treatment requires referral of patients deemed to be at risk to an appropriate physician. A variety of advanced diagnostic methods, including gadolinium-enhanced MRI, Duplex Doppler sonography and angiography are used to confirm carotid stenosis. PMID- 11885438 TI - Drinking water in South Africa: implications for fluoride supplementation. AB - During the extremely heavy rainfall period in the first half of 2000 in certain summer rainfall regions of South Africa a similar investigation as in 1983 1984/1985 was done but on a smaller scale, and compared to the previous results. A greater reduction (P < 0.05) of the fluoride levels in the drinking water than previously recorded has recently been demonstrated, thus strongly accentuating the effect of heavy rains. Presently, for fluoride prescription we recommend that the prescriber should decide whether it is a dry or wet period for his area and then read the fluoride content for that area, from the higher or lower end of the water fluoride content range from the tables published in 1988, 1991 or 1994. When in doubt the higher fluoride level should be used as a measure of safety against overprescribing. If only one value exists it should be used until further notification. An extended study is in process to set a single table for South Africa. PMID- 11885437 TI - Benign mucous membrane pemphigoid--a case report. AB - Benign mucous membrane pemphigoid (BMMP) is a relatively rare, chronic vesiculobullous disease. It frequently affects postmenopausal women, although cases have been reported in younger individuals. Benign mucous membrane pemphigoid has a predilection to affect multiple mucosal surfaces including the gingivae, hard and soft palate, alveolar ridge, nose, pharynx, gastrointestinal tract, genitalia and the conjunctiva. It is seen clinically as bullae or erosions on the mucosae or gingivae. The bullae rupture after 24-48 hours and the erosions heal within 7-14 days, sometimes with scar formation. Scarring frequently occurs with ocular mucosa involvement and may contribute to blindness. In order to make a diagnosis, the clinical features must be correlated with microscopic and immunopathological findings. Benign mucous membrane pemphigoid is treated with high doses of corticosteroids and immunosuppressive agents. This paper is a presentation of a case report in a 36-year-old woman. PMID- 11885439 TI - [The accuracy and consistency of dental radiometers]. AB - Radiometers are used in dentistry to evaluate the intensity of light emitted by curing lights. This article discusses the accuracy and consistency of radiometers. The study was done as two experiments, dividing radiometers by age. In experiment 1, one Heliolux II curing light was tested nine times with each of four old radiometers. In experiment 2 the same curing light was tested with three very new radiometers, under identical circumstances. In experiment 1, the average intensities measured by the radiometers ranged between 262 and 348 mW/cm2, while the standard deviation varied between 7.59 and 42.03. In experiment 2, the average intensities measured by the radiometers ranged between 240 and 283.75 mW/cm2, while the standard deviation varied between 0.00 and 4.63. The seven radiometers differed significantly (P < 5%). The conclusion of this study is that the consistency of the radiometers differs depending on the age of the unit, the state of repair of the unit, and how often it is standardised. In this study it was impossible to evaluate the accuracy of the radiometers. PMID- 11885440 TI - Understanding and cherishing the differences in our culturally diverse nation. PMID- 11885441 TI - "Nurse, my head hurts": a review of childhood headaches. AB - Headache, a frequent occurrence during childhood, can have a number of etiologies. Most headaches are benign, but all require appropriate assessment. Common types include sinusitis, migraine, and muscle contraction (tension). Headache assessment includes both history and physical examination. The headache history consists of the history of present illness, past history, family history, and environmental and social history. Physical examination begins with general observation and vital sign measurement and proceeds to specific inspection of the head, neck, and facial structures. The nursing interventions carried out are dependent on the interpretation of assessment findings. Pharmacologic interventions, commonly acetaminophen or ibuprofen, may be appropriate if pain management protocols exist. Nonpharmacologic strategies for headache relief include reassurance, rest, ingestion of simple and complex carbohydrate foods, relaxation exercises, or home care. Children with migraine headaches benefit from specific interventions, and children exhibiting headache warning signs should have emergency measures instituted. PMID- 11885442 TI - The complicated task of managing school health programs. AB - There is growing awareness of the important link between health and education in our society. Children need to be healthy to learn, and they must learn to be healthy. The 8-component coordinated school health program, developed in the early 1980s, rests on the premise that everybody in a child's environment can contribute something, although no one can address a child's health problems effectively by working alone (Tyson, 1999). There is, however, one essential component missing from the coordinated plan: program management. Many of the components of a comprehensive school health program exist in some aspects in our schools. What is lacking is the coordination of these services. School nurses possess the experience, skills, and knowledge necessary to provide the missing link of a comprehensive school health program and must become leaders in this essential effort to care for children and their families. PMID- 11885443 TI - Medication management problems reported by subscribers to a school nurse listserv. AB - Given the potentially serious consequences of suboptimum medication management practices in elementary and secondary schools and the fact that this topic has been subject to little empirical inquiry, the purpose of this study was to obtain a preliminary understanding of the types of medication management problems that school nurses face, as well as the strategies they use to solve those problems. An analysis of messages related to medication management that appeared on the SCHLRN-L listserv (a 1,400-member discussion group for schools nurses) was performed. All messages sent to the listserv during a 5-month period were monitored. Any discussion thread that began during this time period and addressed the management of medications was selected for possible inclusion in the study. The result was a group of 71 threads. The listserv participants described a wide array of medication management problems and suggested numerous strategies for solving them. This study shows that serious medication management problems exist in schools and that school nurses make use of various strategies or "tricks of the trade" to handle these problems. This research highlights the need for a contribution from the pharmacy profession in this important and neglected area of drug therapy. PMID- 11885444 TI - International students' perceptions of health care. AB - International students are coming to the United States in increasing numbers to study in the independent school setting. Compared with the general student population, fewer international students seek nursing intervention or medical attention for common illnesses and ailments. The purpose of this study was to determine barriers, real or perceived, that impede international students in high schools from seeking and utilizing health services that are available to them in the school setting. This qualitative study used Leininger's cultural care and universality theory. Self-contained focus groups were used to collect data that provided insight into attitudes, perceptions, and opinions as they relate to health care services, practices, and beliefs. Nursing implications include incorporation of culturally appropriate nursing care and the expansion of nursing practice to include traditional homeland remedies and medications in the treatment of international students in the school setting. PMID- 11885445 TI - The health behavior selection process of young adolescents. AB - The health behavior selection process of young adolescents was explored using the grounded theory research method. Data were generated from in-depth interviews, field notes from school settings, and memos. A conceptual model was generated identifying 4 major categories: assessing/valuing, confirming, choosing, and safeguarding. The processes identified in the model offer the potential of developing new strategies for school nurses to influence health behavior outcomes among young adolescents. PMID- 11885446 TI - National school nurse certification. Part II: Questions and answers. AB - The National Association of School Nurses proposed the concept of certification for school nurses in the 1970s. The development and evolution of the school nurse certification process, from concept to reality, were described in the October 2000 Journal of School Nursing (Gregory & Marcontel, 2000). Readers were asked to submit questions to be answered in the second article. This article describes issues and answers questions from school nurses about national certification, gives information from past and current presidents and certified school nurses regarding their experiences, and presents a forecast on the future of school nurse certification. PMID- 11885447 TI - Consent and release. AB - Because school nursing practice by definition involves delivering services to minors, school nurses often find themselves with questions about consent. Because school districts, much like other health care providers, occasionally face liability for the undesired health care outcomes of their clients (i.e., students), another question that surfaces frequently deals with release from liability. Each question must be answered based on analysis of the specific facts and applicable laws. This article provides an overview of the principles that provide school nurses with general guidance on consent issues and release of liability and, by way of example, applies them in the answers to four specific questions forwarded by practicing school nurses. PMID- 11885448 TI - Health Jeopardy: a game to market school health services. AB - Health Jeopardy was developed by members of the School Nurse Organization of Minnesota to present information about health and to market the role of the school nurse. Although it was created for the Minnesota State Fair, it has been used in a variety of community settings. This interactive and fun game has been popular with both children and adults. Contestants were selected from the state fair audience, and each competed for prizes by answering questions about school health under the five categories of first aid and safety, nutrition, immunizations, communicable diseases, and school nurse facts. In addition to the state fair, Minnesota school nurses have used this game as both an informational and promotional tool in their school and community fairs. PMID- 11885449 TI - Lyme disease (LD) is a potential health concern for school-age children. PMID- 11885450 TI - Evolution of saliva and serum components in patients with oral candidosis topically treated with Ketoconazole and Nystatin. AB - The present study involves the analysis of some saliva components (SC) and serum components in patients with oral candidosis topically treated with Ketoconazole 2% (K) or Nystatin 100,000 IU (N). Twenty-four male and female patients, age range 39-82 years, were included in the study. A double-blind study was undertaken in which the patients were divided into 2 treatment groups. These groups were compared with a control group (CG) of 16 healthy patients, both male and female, age-matched with the treated groups. The parameters evaluated were oral mucous membrane lesion index (MLI), CFU of Candida, saliva flow rate, protein-bound Fe (Fe-prot), Fe-prot binding capacity (Fe-prot cap), IgAs, peroxidase activity (PA), hypothiocyanite and thiocyanite. The values of Candida CFU and MLI were significantly reduced in patients treated with K and N. The pre treatment values of SC as compared to the CG revealed a reduction in Fe-prot and Fe-prot cap. These parameters reach values similar to control towards the end of the treatment. The PA was significantly higher in candidosis patients and fell to control values with treatment. The other SC and serum components did not exhibit significant differences with the CG. Patients with oral candidosis treated locally exhibit not only an improvement in clinical manifestations but also a return to control values of altered SC. PMID- 11885451 TI - Dynamics of bone loss in experimental periodontitis. AB - A dynamic histomorphometric study of bone loss in periodontitis induced by inserting a thread ligature around the neck of the lower first molar of Wistar rats weighing 300 g. was performed. Bone formation fronts were labelled twice by tetracycline injection (day 1 and day 14). On day 16 the animals were divided into 4 groups: Two experimental (ligature in place) and two controls (no ligature). Animals in one experimental and one control groups were killed 72 hours post insertion, and the other two groups 96 hours post-insertion. Grinding sections of the first molar were obtained to perform histomorphometric studies on microphotographs taken under fluorescence microscopy. At 72 hours results showed total loss of the double labeling in the mesial wall, partial loss in the top of the crest and no loss in the distal wall. At 96 hours, loss of the double labeling at the top of the crest was greater, while only one label (the first) could be observed in the distal wall. These results show that in this periodontitis experimental model, bone loss is initiated and is more rapid in the bone remodeling (mesial) wall than in the modeling (distal) wall. This understanding of bone loss dynamics enables the characterization of the model employed herein, contributing to further studies on the course of periodontal disease under different experimental conditions. PMID- 11885452 TI - Relation between demographic and epidemiological characteristics and permanency under a dental health care program for HIV infected patients. AB - The association between factors involved in health care and the health status of the people has been proven. The use of health care services, particularly in the case of patients who suffer from chronic pathologies, has been the object of many studies aimed at establishing factors which contribute to guarantee permanence in treatment and implementation of health care controls. The purpose of the present study was to identify the response of HIV infected or AIDS patients to the oral health care program and establish the association between permanence in treatment and the presence of risk factors, epidemiological or demographic conditions of the users. Ninety patients selected at random from the 300 who attended the Clinic for High Risk Patient Care, School of Dentistry, University of Buenos Aires (CLAPAR, Spanish acronym) during 1994-1995 were included in this study. The patients were assigned to one of seven groups, according to their permanence in treatment and commitment to the program during the phase of maintenance in health. Each of these categories was characterized in terms of age, sex, educational level, place of residence (CIRFS, 1990), type of job, type of health coverage and place where healthcare was received, risk behavior and date of positive serological diagnosis. The frequency of each variable was established. Contingency tables were employed to establish the statistical significance of the association between the different variables and the patient categories. The data revealed that 24.2% of the patients performed occasional or emergency consultation, 57.1% achieved discharge with or without the assistance of the social worker or are still in treatment and 18.7% abandoned the program. Significant association were found between the response to odontological treatment and the following variables: place of residence, date of positive serological diagnosis, and risk behavior. We may conclude that certain demographic, epidemiological or life-related factors would be linked to the response to odontological treatment. PMID- 11885453 TI - Caries experience in 3 year old children of Cordoba, Argentina. AB - The results of a study of caries in a 3 year old cohort of children performed by the CLACyD (Spanish abbreviation by initials for Nutrition, Growth and Development Program of Cordoba) are analysed. The prevalence of caries was 19.7% for boys and 16.1% for girls. Dmf-t and dmf-s values were 0.55-0.75% for girls and 0.73-1.22% for boys. The most affected element was the first mandibular molar (16.30% of the population). Extracted and filled elements accounted for 1.8 and 5.9% of the dmf-t index. According to the Caries Analysis System, a higher prevalence of the PF (pits and fissures) pattern and the AM (anterior maxillary bone) pattern was observed as compared to the PP (proximal posterior) and BL (buccolingual) patterns. The percentage distribution of BL lesions over all the patterns exhibited statistically significant (p = 0.05) differences between girls and boys. The PF and PP patterns were more severe in boys (p = 0.0006 and p = 0.002 respectively). Based on the present data we conclude that this population exhibits marked prevalence which can rapidly increase and pose a threat to temporary and mixed dentitions. We herein recommend preventive measures of widespread use (regarding toothbrushing, and eating habits with particular reference to carbohydrate consumption) and preventive-therapeutic treatments for diseased patients (topical applications of fluoride, sealing agents and active treatment of caries). PMID- 11885454 TI - Pulpal response to intrusive orthodontic forces. AB - Many studies have examined the effect of orthodontic forces on supporting tissue. However, their effect on dental pulp has not been sufficiently studied. The aim of the present study was to evaluate, histologically, the pulpal response to the action of intrusive orthodontic forces. The study was performed on 40 first premolars which had to be extracted for orthodontic reasons from 11-17 year-old patients. The twenty teeth of the experimental group received, prior to extraction, an intrusive force with an initial load of 150 g over a period of 15 to 20 days. The control group comprised the 20 homologous teeth which were not submitted to intrusive forces prior to extraction. All the teeth were fixed in 10% buffered formalin, decalcified, embedded in paraffin, sectioned and stained with hematoxylin-eosin. Light microscopy observation revealed alterations in predentine, calcium deposition, fibrohyalinosis, congestion, inflammation and haemorrhage. The data were statistically analysed employing Fisher's exact probability test. Congestion, haemorrhage and fibrohyalinosis were significantly (p < 0.005) greater in the experimental group than in controls. PMID- 11885455 TI - In vitro erosive capacity of some fruit juices and soft or low alcoholic strength beverages on human teeth. AB - The in vitro erosive capacity of different fruit juices and soft or low alcoholic strength beverages (n = 50) on human teeth was examined. The end-point was the amount of calcium and phosphate released into the medium following incubation with the test substance. Overall, the erosive capacity rose as the pH of the products fell. A statistically significant negative correlation was found between the dissolving effect and pH or the modulating action in the presence of acids (r = -0.69282 and -0.63708 respectively; p = 0.0000). Conversely, there was no association between erosive capacity and concentrations of calcium, phosphate or fluoride. Considered overall, sport drinks exhibited the greatest demineralizing effect. Beer had the lowest dissolving action. Most of the drinks manufactured with grapefruit, lime or lemon/lime proved to be more erosive than those prepared with apple or peach. Susceptibility to erosion was virtually the same for all types of teeth and dental surfaces. The cervical portion which harbours the anatomical neck underwent dissolution more readily than the coronary portion (p < 0.001). The erosive capacity of the fruit juices and beverages tested is related to their pH. However, a strict relation between free H+ and demineralizing action was not found, probably due to the influence of other factors not considered in the present study. PMID- 11885456 TI - Effects of colchicine on parotid gland: structural and biochemical studies. AB - The present study involves the analysis of structural, ultrastructural and biochemical modifications induced by colchicine (Col) in Guinea Pig parotid gland. The biochemical studies showed that the highest concentration of soluble proteins occurred 4 hs after colchicine injection. The curve of a-amylase activity in the gland showed an increase in enzyme activity over control at approximately 4 hs post-injection. At 8 hours the rise was even greater. The structural and ultrastructural analyses of the gland revealed that 4 hs post injection colchicine exerts its maximum inhibitory effect on secretion. At this treatment time, the cytoplasm contained granules of various sizes and sharp outlines. Large areas of the cytoplasm exhibited material which resembled the granular content and seemed to result from the fusion of granules. No secretion was found in the lumen of the ducts. The inhibitory effect on secretion would be due to the disassembly of microtubules which would in turn impair transport of the granules to the apical surface of the membrane and eventual exocytosis. We herein propose that the effects of colchicine in the experimental conditions of this study would be largely reversible at 24 hours. At this time, structural, ultrastructural and biochemical features were similar to those of controls. PMID- 11885457 TI - [Awa Seck. Nurse, director and woman leader. Interview by Odile Lamy and Colette Pilon-Bergman]. PMID- 11885458 TI - [A new instrument to measure distress in the operated child]. AB - Having noted the lack of a French language tool to evaluate distress behaviours of children in a day care surgery situation, and after an exhaustive review of the literature on the subject, the author developed a measuring instrument for distress levels in such children. The 1999 version of the Echelle descriptive des comportements de l'enfant opere (EDCEO) is a chart of observations used to measure anxiety, fear and pain in children aged 3 to 10 during the day when surgery is performed. The author describes the instrument, its components and the steps leading to its validation. Suggestions as to its uses in clinical and research situations are also presented. PMID- 11885459 TI - [Nursing education in an era of globalization]. PMID- 11885460 TI - [Migrant patients and foot care]. PMID- 11885461 TI - [Kangaroo mothers in Mexico]. PMID- 11885463 TI - Older women's experience of living with chronic leg ulceration, December 1999. PMID- 11885464 TI - Bone growth in nonorganic nutritional dwarfing rats. AB - Since no data are available to characterize mandibular growth in nonorganic nutritional dwarfing (ND), the purpose of the present study was to describe the effects of a diet on mandible and femur growth in a nutritional dwarfish animal model. Male Wistar rats were divided into two groups of 10 animals each: Control (C) and Experimental (E80: diet-restricted group). C rats were fed a standard diet ad libitum. E80 rats received 80% of the amount of standard diet eaten by group C. Food intake and body weight (Wt) and length (Lt) were recorded periodically. Growth data (Wt and Lt) were expressed as a Z-score of weight-for length (WLZ) ratio, an index of body size. Five animals of each group were selected at random at 4 and 8 weeks and sacrificed. Additionally at t = 0, 5 animals were sacrificed for baseline measurements. Mandibular growth was estimated directly on the right mandible by measuring ten dimensions. Femur growth was estimated from Wt and Lt measurements of the bone. Mandibular weight, area, length and height were negatively affected by dietary restriction during the first 4 weeks of the experimental period. Mandibular growth ceased after this point. Dimensions corresponding to the alveolar unit did not change with time. However, all other dimensions were negatively influenced but not to the same extent. Femur rather than mandibular weight was severely affected. Therefore, the negative effects of the nutritional stress that occurs after weaning would be stronger for the femur, than for the mandible. Femur length was also negatively affected by suboptimal nutrition. In summary, the results of the present study showed that mandible and femur growth respond differently to mild chronic food restriction. These observations could be explained in terms of the different critical bone growth periods and of the time at which nutritional stress was imposed. PMID- 11885462 TI - The Clair Commission. The health system under examination. PMID- 11885465 TI - Effect of photic stimuli on rat salivary glands. Role of sympathetic nervous system. AB - Saliva secretion during feeding facilitates chewing, swallowing and other oral functions. Between meals, a "resting saliva" is elicited to allow speaking and contribute to maintain soft and hard tissues health. Chewing is the main stimulus for "stimulated saliva" secretion. Mouth dryness and other less well known stimuli control "resting saliva". In humans the stimulus of the light increases the parotid saliva flow rate. Saliva secretion occurs in response to a reflex. Both motor branches of the autonomous nervous system drive efferent outputs to the salivary glands. Cellular bodies of sympathetic motor fibers innervating salivary glands are located in the superior cervical ganglia. A multisynaptic pathway couples the superior cervical ganglia to hypothalamic areas related to the control of autonomous and endocrine functions. Projections from suprachiasmatic nuclei involved in circadian rhythms control reach those areas. Salivary glands postsynaptic beta-adrenoceptors control synthesis and secretion of proteins. Postsynaptic alpha 2-adrenoceptors modulate salivary responses mediated by alpha 1 and beta-adrenoceptors. Parotid alpha-amylase circadian rhythm in suckling rats, suggest that the sympathetic nervous system mediates an effect of light on saliva secretion. Analysis of: 1) parotid fine structure, 2) submandibular secretory response to adrenergic agonists, and 3) submandibular 3H clonidine binding to alpha 2-adrenoceptors, demonstrated that an increase of sympathetic reflex activity occurs in salivary glands of rats chronically exposed to constant light. Similar effects were observed in rats chronically exposed to immobilization stress. Catecholamine biosynthetic enzyme mRNA levels in adrenal glands and superior cervical ganglia suggest that changes induced by light on salivary sympathetic reflex activity are mediated by plasma catecholamines released by adrenal glands. Post and presynaptic alpha 2 adrenoceptors could play an important role in saliva secretion control when light or stress stimuli modify the sympathoadrenal system. PMID- 11885466 TI - Enlargement of periosteocytic lacunae associated to mechanical forces. AB - Demineralization of bone has been linked to the action of osteocytes via the process of osteocytic osteolysis. The aim of this study was to assess the influence of mechanical forces produced during orthodontic movements on the osteocytic lacunae. Orthodontic movements were achieved employing a device constructed "ad hoc" for rats that exerts a force of approximately 70 gr. The experimental animals and the corresponding controls were killed 48 and 96 hours after the onset of the experiment. Histologic sections oriented along the bucco palatine axis were employed to measure the area of osteocytic lacunae to infer information on volume in keeping with standard stereological concepts. Regions alongside resorption areas of cortical bone and resting areas of palatine bone were evaluated. Osteocytic lacunae associated to erosive surfaces were rounded and rose markedly in area after the application of the orthodontic force (58.4 +/ 6 mm2). Elongated lacunae were present in relation to resting areas (24.8 +/- 2 mm2). The present study shows an association between the increase in size of osteocytic lacunae and the resorption fronts induced by the application of orthodontic forces. This finding would suggest that the osteocyte would participate in the resorption process of bone submitted to pressure. PMID- 11885467 TI - Endodontic anatomy of the root canals of lower incisors. AB - Ninety lower incisors of known age (18 to 20 yr, 30 to 40 yr and over 50 yr) were grouped into 2 sets of 45 pieces each. Cross sections of the root at 4 levels (coronal, middle and apical thirds and 2 mm from the apex) were employed to study: 1. The shape of the canals 2. The changes in diameter induced by age 3. The correlation at the apex of the maximum diameter of the root canal with the Dzero diameter of endodontic instrument. The results showed that the canal anatomy of lower incisors is not as simple as it appears to be and that it is characterized by the predominance of flat shapes even in the vicinity of the foramen. The statistical analysis showed that the dentin deposits, typically age associated, are influenced by the morphological changes that take place. A true compensation occurs between the different buccolingual and mesiodistal diameters, particularly in the vicinity of the apex, to guarantee irrigation in old age and avoid calcification fo the canal. The correlation between the maximum diameter of the canal at 2 mm from the apex and the Dzero diameter of the instruments evidenced the difficulties involved in adequate apical preparation of these teeth, regardless of age. PMID- 11885468 TI - Study of the crevicular fluid flow rate in smokers. AB - The purpose of this study was to investigate if smoking--a risk factor in periodontal disease-affects the crevicular fluid (CF) flow rate. Twenty-nine dental students were included in the control group--non-smokers- (NS) and 34 in the experimental group--smokers- (S). All subjects were enrolled in a rigorous dental hygiene program (RDHP). The Greene-Vermillion plaque index, and Loe Silness gingival index (GI) were recorded. CF was obtained and measured with the Periotron 8000. These recordings were made before and after the RDHP. The results show that the CF mean flow rate was slightly lower in the S group than in the NS group, for both recordings. The analysis of the relation between the CF flow rate and the GI recorded in the dental surfaces, revealed a significantly lower flow rate in the S group for GI 1 (p < 0.01) and GI 3 (p < 0.05). The difference observed between the S and NS groups, may be due to the vasoconstrictor action of the cigarette components (nicotine and/or metabolites) on the gingival vasculature. PMID- 11885469 TI - Novice nurse educators' lecture room instructional management competence. AB - The lecture room instructional management competence (LRIMC) of novice nurse educators (NNEs) in the Republic of South Africa (RSA) was investigated by means of a quantitative exploratory survey using questionnaires. The findings indicated that NNEs benefited from mentors' guidance, experienced reality shock on entering their first teaching situation, and lacked LRIMC--according to the perceptions of the NNEs themselves, their students and their mentors. NNEs could benefit from effective orientation programmes and from ongoing in-service education programmes as well as from the availability of mentors assigned to specific NNEs. PMID- 11885470 TI - Selection of magister learners in nursing science at the Rand Afrikaans University. AB - Selection of learners implies that candidates are assessed according to criteria with the purpose of selecting the most suitable learners for the course. A magister qualification is on level 8A of the National Qualifications Framework (NQF). The purpose of a magister qualification in Nursing is the development of advanced research, clinical, professional, managerial, educational, leadership and consultative abilities (knowledge, skills, values and attitudes) for the promotion of individual, family, group and community health. From the above introduction it becomes clear that there is a high expectations of a person with a magister qualification. Such a person should be a specialist, scientist, leader and role model in the profession. A magister programme is human-power intensive as well as capital intensive for both the learner and higher education institutions. It is therefore important to select learners with the ability to achieve the outcomes of the programme. Limited research has been conducted on the selection of post graduate learners. This leads to the question whether the current selection criteria (undergraduate mark and the mark in Research Methodology) are reasonable predictors of success for the magister programmes. In order to answer this question, hypotheses with the following variables were formulated. Achievement/success in the magister programme as reflected by The mark for the dissertation or mini-dissertation. The level of input by the supervisor during the magister programme. The quality of the research article reflecting the research in the magister programme. Undergraduate mark Mark for Research Methodology In order to test the hypotheses a quantitative correlation design was used incorporating documented data of 74 magister graduates. Descriptive and inferential data analysis (Pearson's correlation coefficient, ANOVA and multivariate test) were used. The findings showed Research Methodology to be the best indicator of success in the magister programmes. PMID- 11885471 TI - Errors in anthropometric measurements in neonates and infants. AB - The accuracy of methods used in Cape Town hospitals and clinics for the measurement of weight, length and age in neonates and infants became suspect during a survey of 12 local authority and 5 private sector clinics in 1994-1995 (Harrison et al. 1998). A descriptive prospective study to determine the accuracy of these methods in neonates at four maternity hospitals [2 public and 2 private] and infants at four child health clinics of the Cape Town City Council was carried out. The main outcome measures were an assessment of three currently used methods namely to measure crown-heel length with a measuring board, a mat and a tape measure; a comparison of weight differences when an infant is fully clothed, naked and in napkin only; and the differences in age estimated by calendar dates and by a specially designed electronic calculator. The results showed that the current methods which are used to measure infants in Cape Town vary widely from one institution to another. Many measurements are inaccurate and there is a real need for uniformity and accuracy. This can only be implemented by an effective education program so as to ensure that accurate measurements are used in monitoring the health of young children in Cape Town and elsewhere. PMID- 11885472 TI - Satisfaction with personal and environmental quality of life: a black South African informal settlement perspective. AB - A study was conducted with 487 black adult residents of a South African informal settlement (151 men and 336 women) to ascertain satisfaction with personal and environmental quality of life. It was hypothesised that: (1) health status and life satisfaction were the underlying dimensions of personal quality of life (PQOL); (2) health status and life satisfaction were more strongly associated with PQOL than environmental quality of life (EQOL); and (3) life satisfaction and satisfaction with EQOL were positively related. Seventy per cent of respondents rated their health as good or better. Age, schooling and employment status were significantly related to health, life satisfaction and PQOL. Reliability (internal consistency) coefficients were 0.77 for the 5-item life satisfaction scale and 0.82 for the 12-item EQOL measure. Factor analysis showed that safety and security was the major unmet service need. Health status and life satisfaction explained 38% of the variance in PQOL; health status explained only 4% of the variance in EQOL. Life satisfaction was significantly related to EQOL (r = 0.16, p = 0.01). The results provided support for all three hypotheses. It was concluded that the life satisfaction and EQOL measures had good reliability; there was a definite need for a safety and security programme; and good health was a more important predictor of PQOL than EQOL. PMID- 11885473 TI - The image of the nursing profession as perceived by the community members of three adjacent residential areas of Empangeni in KwaZulu-Natal. AB - The study was undertaken out of concern that nurses and the nursing profession project a negative image to the public they serve. Diverse aspects of nursing examined in this study included standards or quality of nursing care and its influence on encouraging clients to utilise health care services, communication, attitudes, expertise, availability at all times and patients'/clients' participation in decision-making. Nurses are part of the communities they serve. The rationale for the study was that nurses should be alert to perceptions of the communities about the service they provide, whether expressed formally or informally. Survival of nursing as a profession is dependent upon the positive impact it has on consumers in the past, present and the future. The aim of the study was to discover, through a systematic, scientific inquiry, the positive and negative perceptions that communities have about nursing. An exploratory, descriptive study was done in KwaZulu-Natal on a sample of 50 participants from three adjacent historically Black residential areas (townships) using questionnaires. Findings of the study were contrary to the assumption that nursing has a negative image. All aspects except one were rated very good or good by the majority of the participants. Those who had negative perceptions, though in the minority, highlighted important reasons directed at both the authorities and the nurses. These were used as a basis for recommendations for further improvement of the image of nursing. The area of gross dissatisfaction among the majority was feeling unsafe with nurses getting increasingly involved in unionism, which would lead to abandonment of patients in times of industrial action. PMID- 11885474 TI - Tobacco use among black South African university students: attitudes, risk awareness and health locus of control. AB - OBJECTIVES: To provide data on African/black South African university students' tobacco use status, belief in the benefits to health of not smoking, risk awareness in terms of knowledge of the links between smoking and disease, health locus of control, value for health, subjective health status and well-being. DESIGN: Cross sectional. SETTING: University of the North. SUBJECTS: 793 Black University students from non-health courses chosen by random sampling, of these 370 (46.7%) were males and 423 (53.3%) were females in the age range of 18 to 25 years (M age 21.0 years, SD = 3.48). MAIN OUTCOME MEASURES: A measure of smoking, the Multidimensional Health Locus of Control Scale, the Health as a Value Scale, and a measure for subjective health and subjective well-being. RESULTS: The average prevalence of current tobacco use was 15% in men and 1% in women. The proportion of tobacco users who were classified as light users (1-10 per day) averaged 10% in men and 1% in women. Age and being male were significantly positively associated with status and frequency of tobacco use. Awareness of the link between smoking and lung cancer was high (93%), but awareness of the role of smoking in heart disease was very low (16%). The importance to health of not smoking was associated with smoking status (non-smoking versus smoking). Overall, 75% of the current smokers stated that they would like to reduce the amount they smoked. Poor subjective health status and low subjective well-being was associated with smoking status. No significant differences were found among non tobacco users and tobacco users in relation to the three subscales of the Health Locus of Control (Internal, Chance, and Powerful others) and Value for health. CONCLUSION: For about 9% of the male students investigated, a high risk exists to become regular tobacco users for the next 30 years. PMID- 11885475 TI - Concepts and treatment for diabetes among traditional and faith healers in the northern province, South Africa. AB - The purpose of the study was to assess the concepts and treatment modalities for diabetes among traditional and faith healers in the Northern Province in South Africa. The sample consisted of 50 traditional healers (13 females and 37 males) and 50 faith healers (12 females and 38 males). They were interviewed on local terminology, clinical manifestations, causes, curability, and treatment for diabetes, help-seeking behaviour of diabetes patients, and the healers' sources of information about diabetes. Results indicate that all healers were familiar with "diabetes", however, not all of them had seen patients suffering from diabetes. The perceived causes of diabetes by both traditional and faith healers could be divided into (1) diet (especially too much of sugar), (2) heredity, (3) supernatural, and (4) psychological causes. Most traditional healers (92%) and faith healers (90%) indicated that diabetes is curable. Treatments used by the healers in this study included the use of prayer, diet, and herbs. The authors conclude that the concepts and treatment modalities for diabetes among traditional and faith healers should be taken note of by health workers while developing health education programmes in the Province. PMID- 11885476 TI - Exploring the critical care nurses' experiences regarding moonlighting. AB - While moonlighting is so prevalent amongst critical care nurses, there are no documented facts in this country about how it affects the nurses and the hospital management, considering the nature of their work that is both physically, mentally and emotionally strenuous. The aim of this study was to explore the critical care nurses' rationale and experiences regarding holding a second job (moonlighting). A non-experimental exploratory study was done using focus groups. Many positive and less positive experiences were revealed, for example, economical, educational, and psychosocial ones. Participatory control of moonlighting activity was suggested involving both management and staff at functional level. PMID- 11885477 TI - Life stories of families with a terminally ill child. AB - Family units with a terminally ill child have a tendency to withdraw and this isolation may lead to problems in their mental health. A tendency with psychologists, clergy and helpers from other professions is to act as ideal experts on the lives of saddened people. From painful personal experience, this does not seem to enable acquiescence. Therefore, the aim of research on families with terminally ill children, was to explore and describe their lives and to develop an approach to facilitate their families to obtain acquiescence. In this article however, attention will be given to the life-world of families with terminally ill children. The research consists of two phases. In phase one the experiences of four families with terminally ill children are explored and described by means of phenomenological, unstructured, in-depth interviews. In phase two an acquiescence approach, which was designed for educational psychologists to facilitate families with terminally ill children to achieve acquiscence, is described. This approach is based on results from phase one. This article focuses on phase one. In this phase four families were interviewed individually, in the privacy of their homes. The interviews were audiotaped, and were transcribed for the purpose of data gathering. The data was analysed according to Tesch's method and a literature control was performed to verify the results. Guba's model for the validity of qualitative research was used. Five recurrent themes were identified: 1. Families are able to choose their reactions to the crises of having a terminally ill child. 2. When there is a terminally ill child in the family, the family's values change. 3. Acceptance of the circumstances with a terminally ill child, makes life easier. 4. As families with a terminally ill child learn to live every moment to the full, their quality of life improves. 5. As people learn to accept support, their quality of life with a terminally ill child improves. The research indicated that families with terminally ill children move through a lonely and painful process, which is characterised by growth at the end. This growth implies that the life skills mentioned above, were obtained after years of unimaginable suffering. In order to reduce this period of suffering, an acquiescence approach was designed for educational psychologists to facilitate discovery and acceptance regarding the above life skills with family units and thus allow them to achieve acquiescence. PMID- 11885478 TI - Perceptions of epilepsy among black students at a university in South Africa. AB - The present study sought to investigate the relationship between familiarity, attitudes, causative and treatment beliefs about epilepsy in a sample of black young adults (university students) in South Africa. The sample included a convenient sample of 253 second year social science students, 98 (38.7%) males and 155 (61.3%) females in the age range from 18 to 42 years (M = 25.0 years, (SD = 4.2). The questionnaire administered to students in a class room situation included sociodemographic data, and sections on familiarity, beliefs about cause and treatment, sources of information, attitudes about epileptics, and prevention of epilepsy. Results showed that the majority of students, in particular from a rural background, were familiar with epilepsy. Health care institutions were the most important source of information on epilepsy, especially for women. Those who believed in traditional causes of epilepsy also endorsed traditional treatment for it, though they did not see such treatment as curative. Those who believed in a medical treatment did however see such treatment as curative. Although the majority of the students had a positive attitude towards epileptics through sharing a meal or room and willingness to marry an epileptic, 17% thought that epileptics can infect others with their saliva during a seizure, 12% felt an epileptic is a witch or wizard, and about 10% said an epileptic must be isolated. Findings should be included in educational programmes for young adults. PMID- 11885479 TI - System requirements for a computerised patient record information system at a busy primary health care clinic. AB - A prototyping approach was used to determine the essential system requirements of a computerised patient record information system for a typical township primary health care clinic. A pilot clinic was identified and the existing manual system and business processes in this clinic was studied intensively before the first prototype was implemented. Interviews with users, incidental observations and analysis of actual data entered were used as primary techniques to refine the prototype system iteratively until a system with an acceptable data set and adequate functionalities were in place. Several non-functional and user-related requirements were also discovered during the prototyping period. PMID- 11885480 TI - Graduates' perceptions of their midwifery training during the four year comprehensive nursing diploma. AB - Since its inception in 1986, the Comprehensive Nursing Diploma has received many criticisms from registered nurses, who graduated from the pre-existing programmes. This study attempts to examine the perceptions of graduates from this training programme, towards their midwifery education in terms of its adequacy in preparing them for midwifery practice. Twenty-nine graduates of the four year Comprehensive Nursing Programme completed self-administered questionnaires. Data was analysed by means of descriptive and inferential statistics, namely the paired t-test and the Wilcoxon test for matched pairs. Qualitative data was analysed to determine emerging themes and patterns. Differences in competence ratings on entry into midwifery units and those a year later, were significant at p = 0.0001. It was found that graduates appear to have a positive perception of the midwifery component of their training programme. They found the theory aspect to be more than adequate in preparing them for their professional roles. However, with regards to clinical exposure, it was felt that the time period was too short and needed to be extended in order for them to attain clinical proficiency. PMID- 11885481 TI - Psychiatric research in South Africa: a systematic review of Medline publications. AB - BACKGROUND: There is debate about the future path that medical and psychiatric research in South Africa should take. In particular, there have been calls to make research more relevant to the needs of the population. There is, however, little systematically collected data on the nature, strengths, and flaws of past psychiatric research in this country. METHODS: We undertook a MEDLINE search to gather all manuscripts that fell under the umbrella of psychiatric research and published by South Africa-based authors during the years 1966-1997. Several kinds of data were collated from each of the articles, including information about the authors and the journal, as well as information on the focus and type of article. RESULTS: While publications from South Africa continue to grow in number, relatively few involve collaborative research groups and few authors write more than one paper. Many papers relevant to psychiatry were published in general medical journals and many were from general medical departments. While blacks and females have been included in research, a number of important areas have received little attention. CONCLUSIONS: Psychiatry research in South Africa requires additional fostering, including additional resources for research training and arguably additional development of subspecialty focuses. Given the limited resources, and the nature of modern research, increased emphasis on collaboration seems advisable. A number of areas in psychiatry deserve particular attention from future researchers. PMID- 11885482 TI - Osteoporosis: a treatable disease. PMID- 11885483 TI - Prevention of restenosis after percutaneous coronary intervention: the continuing challenge. AB - Percutaneous coronary intervention with angioplasty and stenting is well established in the treatment of coronary artery disease. However, the many advances in technique and equipment over the last couple of decades have yet to significantly reduce the incidence of restenosis. This Achilles' heel has necessitated frequent re-interventions and also introduced a new iatrogenic disease of in-stent restenosis. Brachytherapy and coated stents may be the answer to this difficult problem. Many papers have been published in the last few years on these two new modalities of treatment, and we review the evidence available so far. Early results show that brachytherapy significantly reduce the incidence of restenosis when used in restenotic lesions, and coated stents significantly reduce restenosis in de novo lesions. This early promise of brachytherapy and coated stents, if confirmed in longer-term studies, will represent a breakthrough in the battle against restenosis and may dramatically change the practice of interventional cardiology in the near future. PMID- 11885485 TI - Two case reports on incessant left ventricular tachycardia: curative therapy with radiofrequency ablation. AB - INTRODUCTION: Incessant ventricular tachycardia is a rare arrhythmia which can be life threatening. Treatment with anti-arrhythmic agents may occasionally fail. CLINICAL PICTURE: We report 2 cases of incessant ventricular tachycardia. The first case was a young man with idiopathic left ventricular tachycardia who was in incessant ventricular tachycardia despite treatment with multiple anti arrhythmic drugs and developed dilated cardiomyopathy. The second case was an asymptomatic girl with the incidental finding of an incessant ventricular tachycardia which originated from the left ventricular outflow tract. TREATMENT AND OUTCOME: Both patients underwent electrophysiologic study and radiofrequency ablation with complete termination of the tachycardia. CONCLUSION: Radiofrequency catheter ablation in experienced centres should be the first-line therapy for incessant ventricular tachycardia. PMID- 11885484 TI - Embolisation of a renal artery pseudoaneurysm in a patient with renal malrotation and chronic aortic dissection. AB - INTRODUCTION: Renal artery pseudoaneurysms may arise as a complication of percutaneous nephrolithotomy (PCNL). Prompt recognition and treatment is essential to arrest haemorrhage which may be life threatening. CLINICAL PICTURE: A patient with chronic aortic dissection and malrotated right kidney underwent PCNL for right renal calculus. He developed delayed gross haematuria. TREATMENT: Angiography showed a pseudoaneurysm arising from one of two right renal arteries, which in turn arose from the false lumen of the aortic dissection. The supplying artery was successfully embolised. CONCLUSION: Renal artery pseudoaneurysms can be successfully treated with prompt angiography and embolisation, even in the presence of renal malrotation and aortic dissection. PMID- 11885486 TI - A case report of neurologically unstable fracture of the lumbosacral spine in a patient with ankylosing spondylitis. AB - INTRODUCTION: Fracture/dislocation is uncommonly reported in ankylosing spondylitis involving the lumbosacral spine. CLINICAL PICTURE: We report an 18 month follow-up of a case of neurologically unstable traumatic fracture of the lumbosacral spine in ankylosing spondylitis. TREATMENT/OUTCOME: Posterior decompression, alar-transverse fusion and instrumentation were performed. Anterior diskectomy and fusion were done 6 weeks later. There was solid bony fusion on follow-up and the patient had improvement of 2 Frankel grades and was able to ambulate. CONCLUSION: Combined approaches and longer fixations to stabilise the spine may be required. In the lumbosacral spine, this poses a problem vis-a-vis limited levels of fixation in the sacrum. PMID- 11885487 TI - Bronchopulmonary dysplasia and chronic lung disease of infancy: strategies for prevention and management. AB - Remarkable advances in the treatment of neonatal respiratory disorders, such as antenatal glucocorticoid therapy, surfactant replacement therapy and alternative modes of ventilation, have reduced neonatal mortality and acute respiratory morbidity. However, bronchopulmonary dysplasia and chronic lung disease of infancy remain a substantial complication, especially among the most immature infants. The pathogenesis of chronic lung disease is complex and multifactorial. Prevention and treatment will require a comprehensive multiprong approach with specific interventions and practices focused on different levels of the pathways leading to chronic lung changes. Future improvements in care will require a better understanding of lung development and lung repair mechanisms. However, the ultimate and most effective approach should be a relentless pursuit for measures to prevent premature births. PMID- 11885488 TI - Re: 188rhenium-TDD-lipiodol in treatment of inoperable primary hepatocellular carcinoma--a case report. PMID- 11885489 TI - Percutaneous vertebroplasty in the management of osteoporotic vertebral compression fractures: initial experience. AB - INTRODUCTION: Vertebral compression fractures related to osteoporosis may cause persistent pain which impairs mobility and reduces the quality of life. Percutaneous vertebroplasty is a therapeutic interventional radiology procedure which is used in the management of pain relief in such fractures. It involves the injection of bone cement [polymethylmethacrylate (PMMA)] into the collapsed vertebrae under radiological guidance. This provides pain relief as well as increases the strength and stability of the vertebra. MATERIALS AND METHODS: A total of 16 patients with 17 osteoporotic compression fractures which were treated with percutaneous vertebroplasty over an 18-month period were studied. There were all women with the exception of 1 male patient. Their ages ranged from 61 to 87 years. The fracture sites were at the thoracolumbar junction from T12 to L3 levels. The majority of cases only required a unipedicular injection, with bipedicular injections in 3 cases. All cases were performed in the angiographic suite in the radiology departments, with biplanar fluoroscopy in one hospital. PMMA was injected in a semi-solid state under radiological guidance and screening into the collapsed vertebrae. RESULTS: All cases showed good technical success with no mortality or major complications. Only 2 cases had minor complications of cement leakage into the soft tissues of the back and adjacent disc space, respectively. There was sufficient pain relief in all patients and they were well enough to be discharged within 1 to 5 days after the procedure. Patients were followed up to evaluate the degree of long-term pain relief as well as analgesic usage. CONCLUSION: Percutaneous vertebroplasty is a new and minimally-invasive modality of treating pain in patients with osteoporotic compression fractures who are refractory to medical therapy. Under adequate imaging guidance, the risks of complications are minimal while the potential benefit to patients and their care givers are significant. PMID- 11885490 TI - To establish the normal bone mineral density reference database for the Singapore male. AB - INTRODUCTION: The objective of this study was to establish the normal bone mineral density (BMD) reference curve for the Asian Singapore male. MATERIALS AND METHODS: Three hundred and eighty-three male subjects were enrolled; comprising of 309 Chinese, 44 Malays and 30 Indians resident in Singapore. Bone mineral density was measured at the lumbar spine and left hip using a Hologic QDR 4500 Elite dual-energy X-ray absorptiometry (DXA) scanner. RESULTS: The mean peak BMD for the average lumbar spine and the neck of femur was 1.006 g/cm2 and 0.97 g/cm2, respectively. The mean peak BMD was taken at the 20 to 24 years age group at both the hip and spine based on data distribution for the various age groups. The BMD corresponding to -2.5 standard deviations from the peak adult value was 0.719 g/cm2 for the average lumbar spine and 0.655 g/cm2 for the neck of femur. CONCLUSION: This Asian male BMD reference database, which is 10% and 5% lower than corresponding values from the Caucasian reference database, allows for more accurate diagnosis of osteoporosis in Asian males. PMID- 11885491 TI - An Asian perspective to the problem of osteoporosis. AB - INTRODUCTION: Most research into the problem of osteoporosis has been carried out in Caucasian populations. This review highlights emerging data from research on osteoporosis among Asians, and some differences from Caucasian data. METHODS: A non-systematic review of the English-language literature on various aspects of osteoporosis among Asian populations was carried out. RESULTS: Compared to Caucasian populations, epidemiological trends in Asian populations appear to be developing along similar lines, although rates of hip fracture do differ by country and ethnicity, and are generally lower. Bone mineral density (BMD) carries similar relevance with regard to fracture risk, although hip geometry is also believed to have some impact. Risk factors for osteoporosis and fractures are somewhat similar, although dietary factors seem to play a larger role as reported in Asian studies. A uniquely Asian self-assessment tool based on clinical factors has been developed to assist in case-finding of osteoporotic patients. The few intervention trials with hormone replacement, alendronate and parathyroid hormone appear to show similar responses between Asians and Caucasians, although the response to vitamin D analogs in Asians appears better. Some differences in gene polymorphisms between Asians and Caucasians exist, and these may impact on BMD and fractures via different gene-environment relationships. CONCLUSIONS: Many aspects of osteoporosis in Asia appear similar to the West, but several interesting differences have emerged. These might lead to refinements in the strategies to manage osteoporosis within the Asian context. PMID- 11885493 TI - Osteoporosis in relation to menopause. AB - With an ageing population in Singapore, it is anticipated that postmenopausal osteoporosis and related fractures will be an increasingly important health issue in the coming decades. Oestrogen replacement therapy has a long history of use to treat postmenopausal problems including osteoporosis. The availability of other potent agents such as bisphosphonates, calcitonin and selective oestrogen receptor modulators has enriched the therapeutic options. This article reviews the evidence for the choice of appropriate agents, and strategies to prevent and treat postmenopausal osteoporosis. PMID- 11885492 TI - Osteoporotic hip fractures in Singapore--costs and patient's outcome. AB - INTRODUCTION: Little data are available on costs and outcome associated with osteoporotic hip fractures in Singapore. MATERIALS AND METHODS: A retrospective study was carried out on 280 consecutive hip fractures in patients older than 60 years admitted over a 3-year period. RESULTS: The mean age of patients was 80 years. Sixty-eight per cent were female and 58% were intertrochanteric fractures. Two hundred and sixty-four patients (95%) were operated upon. The mean total hospitalisation period was 17 days. Seventy-six per cent were staying in their own homes prior to the hip fracture while 22% were admitted from nursing homes. After surgery, 63% of patients returned to their homes while 26% needed nursing home care. The index admission mortality rate was 5.7%. Mortality was 26% at 1 year. Of those alive at 1 year, ambulatory status was: 28% were walking without aids, 39% were walking with aids, 24% were wheelchair bound and 9% were bedridden. Poor ambulatory function at discharge was related to increased mortality at 1 year. The average cost incurred was S$7367. The average government subsidy amounted to 82%. Ninety-one per cent of patients were warded in subsidized beds. Breakdown of cost was as follows: hospital stay, 42.6%; surgery, 36.5%; ward treatment, fee 9%; laboratory and X-ray investigations, 4.4%; implant costs, 3.5%; drugs, 1.6% and rehabilitation, 1.1%. Multivariate analysis showed that the cost is significantly related to days spent awaiting surgery, preoperative sepsis, operative complications and cerebrovascular accidents. Young age, good American Society of Anesthetists (ASA) status and endoprosthesis replacement were factors that allowed for early ambulation and lower costs. CONCLUSION: The mortality rates and functional outcome are not very different from published studies in the West. More of our patients returned to their own homes after hospitalisation. Early surgery, close involvement of the medical social worker and intensive physiotherapy or provision of outpatient therapy facilities may help cut cost of treatment. PMID- 11885494 TI - Osteoporosis risk factor assessment and bone densitometry--current status and future trends. AB - INTRODUCTION: Osteoporosis and fragility fractures are problems which will increase in significance as the population of the elderly in many countries increases. The availability of bone mineral density (BMD) measurements, which can define osteoporosis, allows the implementation of effective therapeutic interventions to those at risk for fractures before they occur. Because of the increasing at-risk population and the relatively high cost of these measurements and interventions, a case-finding strategy to detecting osteoporosis has been widely recommended. This review highlights the approach to detecting and diagnosing osteoporosis. METHODS: A non-systematic review of English-language literature on the diagnosis and assessment of osteoporosis was conducted. RESULTS: Many risk factors have been found to be associated with osteoporosis and fractures. These risk factors may be utilised for case finding in deciding who should be evaluated for osteoporosis. Clinical self-assessment tools have been developed to identify women likely to have low BMD who might be recommended for BMD. Several techniques are available for BMD measurement, but the technique of choice for the diagnosis of osteoporosis is dual-energy X-ray absorptiometry (DXA) measured at the hip. BMD thresholds have been widely used to guide osteoporosis treatment. However, recent research has been directed at using risk factor assessment and self-assessment tools to derive medium-term fracture risk as a guide to therapeutic intervention. CONCLUSIONS: Proper selection of individuals for evaluation and treatment for osteoporosis would include risk factor assessment and appropriate BMD measurement to determine the risk of fracture and the need for intervention. PMID- 11885495 TI - Medical treatment of osteoporosis--increasing options. AB - INTRODUCTION: Many drugs are now available for the prevention and treatment of osteoporosis. They have differing levels of evidence of efficacy and each may be used according to clinical indications. METHODS: A Medline search of clinical drug trials using various therapeutic agents used for osteoporosis was carried out. RESULTS: Several randomised controlled trials have been carried out using many agents. The agents with the best data to date with regards to the prevention of spine as well as hip fractures in patients with prevalent fractures belong to alendronate and risedronate. Parathyroid hormone has been shown in one trial to reduce the risk of non-vertebral fractures. For reduction of spine fractures, in addition to the above two bisphosphonates, many agents, in particular raloxifene, have been shown to be clearly beneficial. Weaker data exist for hormone replacement, calcitonin, cyclical etidronate and the vitamin D analogues calcitriol and alfacalcidol. CONCLUSION: There are many therapeutic agents shown to be clearly effective in the treatment of osteoporosis. PMID- 11885496 TI - Steroid-induced osteoporosis. AB - INTRODUCTION: Corticosteroids have major effects on calcium metabolism, leading to accelerated osteoporosis and fracture. METHODS: This review will attempt to summarise current knowledge about their effects in light of new information and important remaining questions, especially with respect to management of this common condition. RESULTS: Corticosteroids affect bone through multiple pathways, influencing both bone formation and bone resorption. Evidence from randomised trials suggests that postmenopausal women receiving corticosteroids are at greatest risk of rapid bone loss and consequent fracture and should be actively considered for prophylaxis. Based upon available evidence, the rank order of choice for prophylaxis would be a bisphosphonate followed by a vitamin D metabolite or hormone replacement. CONCLUSIONS: With early therapy, corticosteroid bone loss can be effectively prevented or reversed. PMID- 11885497 TI - Bone fragility in Asian and Caucasian men. AB - Hip and vertebral fractures are a public health problem in men of Asian and Caucasian origin. Inferences regarding gender and racial/ethnic differences in fracture rates must be made cautiously as problems in case ascertainment and classification of hip fractures, and problems in defining what constitutes a vertebral 'fracture' have not been solved. However, methodological issues probably do not entirely account for the heterogeneity of fracture patterns. There is likely to be a wide variation in fracture rates from country to country in Asia as reported in studies in Europe. The reasons for this heterogeneity are unknown. Caucasian men lose similar amounts of bone as Caucasian women during ageing from the endosteal surface of the bone. Net bone loss is less in men than women because men form more periosteal bone during ageing than do women. The extent of periosteal and endosteal bone modelling and remodelling have not been studied in Asian men and women. Nor have there been hypothesis-driven studies designed to compare periosteal apposition and endosteal bone loss in Asian males compared to Caucasian males. Sex hormone deficiencies contribute to abnormalities in skeletal size and mass during growth, remodelling imbalance and bone loss during ageing in men. The larger peak bone size and greater periosteal apposition with ageing in men compared to women is most likely to be androgen-dependent in Caucasians and Asians. Androgen deficiency may also partly account for reduced bone formation and negative bone balance at the basic multicellular unit (BMU). Oestrogen deficiency during growth is associated with reduced bone mass and increased leg length in males and females. Oestrogen deficiency during ageing may account for trabecular bone loss in men by increasing remodelling rate. There have been no anti-fracture efficacy studies done in Asian males. Studies on the pathophysiology of osteoporosis in males have given insight into the pathophysiology of osteoporosis in females. Similarly, collaborative research efforts between groups around the world will facilitate comparative studies in Asian and Caucasian communities. The results of this work will provide important insights into the pathophysiology of bone fragility in both groups. PMID- 11885498 TI - Osteoporosis--a worldwide problem and the implications in Asia. PMID- 11885499 TI - The Bone and Joint Decade 2000-2010: for prevention and treatment of musculoskeletal disease. AB - The Bone and Joint Decade 2000-2010 has been established to increase awareness of the scale and impact of musculoskeletal disorders on the individual, health care systems and the society. It is a multi-disciplinary initiative involving professional bodies, patient care groups, research organisations and the community. PMID- 11885500 TI - Clinical update on osteoporosis. AB - Osteoporosis and fractures are an increasing problem in many countries. Continued research into the genetics and molecular pathophysiology of osteoporosis improve our understanding of this disease, and may provide insights into novel strategies or therapeutics which might be developed. Increased awareness of the epidemiology of osteoporosis, risk factors, and the availability of methods to quantify skeletal integrity such as bone densitometry, allow the condition to be anticipated and diagnosed before fractures occur, and facilitate measured decisions about further evaluation and appropriate management. As with most chronic diseases, healthy lifestyle measures are recommended. In addition, pharmaceutical options which have been shown to be effective in reducing fracture rates among those with or without fractures are increasingly available. The decision to institute therapy and choice of drug should ideally be rational and individualized. PMID- 11885501 TI - Single centre review of radiologically-guided percutaneous nephrostomies: a report of 273 procedures. AB - INTRODUCTION: To evaluate the technical success and complications associated with radiologically-guided percutaneous nephrostomies (PCNs) in a single centre. MATERIALS AND METHODS: A total of 273 PCNs performed in 190 patients in our hospital over a 3-year period from January 1997 to December 1999 were retrospectively reviewed. The study population consisted of 97 males and 93 females, ranging in age from 13 to 91 years. The main indications were urinary obstruction (77.7%), pyonephrosis (18.3%) and urinary diversion (4%). Demographic variables, technical and risk factors related to the procedure, complications, effect on urine cultures and body temperature; and subsequent patient management were examined. RESULTS: The technical success rate was 99%. The 30-day mortality was 7.2%, none of which were procedure related. Haemorrhage requiring transfusion occurred in 4.3% while septicaemia affected 3.2% of patients. Drainage catheter complications included catheter dislodgement and blockage which were 11.9% and 4.1%, respectively. Thirty-one per cent of PCNs subsequently underwent ureteric stenting as the definitive treatment modality. CONCLUSION: Radiologically-guided PCN is a safe procedure with a high technical success rate. PMID- 11885502 TI - Differentiation of malignant vertebral collapse from osteoporotic and other benign causes using magnetic resonance imaging. AB - INTRODUCTION: Elderly patients presenting with backache and vertebral collapse are a diagnostic challenge. Plain X-rays, computed tomography and radionuclide bone scans have not always reliably distinguished between benign and malignant causes. Magnetic resonance imaging (MRI) may be able to do so. MATERIALS AND METHODS: Patients who underwent MRI evaluation for vertebral collapse were retrospectively studied. Over a 21-month period from January 1995 to September 1996, 47 patients with 58 vertebral collapses were studied. Benign and malignant aetiologies were established by serial imaging, clinical outcome and histology. Imaging was performed with T1 and T2-weighted sequences, with contrast enhancement in some patients. Collapsed vertebrae were examined for appearance of marrow on T1 and T2-weighted sequences and after contrast administration, signal intensity of adjacent discs, degree of marrow involvement, involvement of posterior elements, presence or absence of paraspinal mass and end-plate integrity. Agreement between the final and radiological diagnosis was evaluated. RESULTS: There were 36 benign vertebral collapses (20 osteoporotic, 7 post traumatic, 9 infective) and 22 malignant ones (20 metastatic carcinoma, 2 multiple myeloma). Features which pointed to malignant cause were hypointense marrow on T1-weighted images, marrow enhancement after intravenous contrast, greater than 50% marrow involvement and involvement of posterior elements. Of the vertebral collapses due to infection, 78% showed end-plate disruption. CONCLUSIONS: This study shows that MRI can be used to accurately differentiate between benign and malignant causes of vertebral collapse. Further differentiation between an osteoporotic, traumatic or infective cause can be done with the help of clinical history and evaluation of end-plate integrity. PMID- 11885503 TI - Intussusception: a three-year review. AB - INTRODUCTION: Intussusception is the commonest cause of intestinal obstruction in infants and young children. MATERIALS AND METHODS: This report reviews the clinical presentation, investigations and outcomes of patients with intussusception treated at the KK Women's and Children's Hospital between 1 May 1997 and 30 April 2000. RESULTS: The study population comprised 160 consecutive patients treated for intussusception at our hospital over this 3-year period. The commonest symptom was vomiting; present in 135 patients (84.4%). A palpable abdominal mass was present in 90 patients (56.3%). The classical features of vomiting, abdominal pain, abdominal mass and rectal bleeding were present together in only 12 patients (7.5%). Abdominal ultrasonography was performed in 155 patients. One hundred and fifty-two patients (98.1%) had the classical target lesion on ultrasonography. Air enema reduction was attempted in all except 6 patients. In the majority of patients (130 or 84.4%), the intussusception was reduced successfully by air enema reduction. There was no association between the duration of symptoms before radiological reduction and the outcome of radiological reduction. CONCLUSIONS: As the four classical features of intussusception were present together in only 7.5% of our patients, a high index of suspicion is necessary when any of the signs and symptoms are present in an infant or young child. Abdominal ultrasonography is the diagnostic investigation of choice. Air enema reduction was successful in 84.4% of patients and the duration of symptoms did not reduce the success rate. Thus, air enema reduction should be attempted in most patients unless they have absolute contraindications. PMID- 11885504 TI - Epidemiology of beta-haemolytic group G streptococcal bacteraemia in Singapore (1996 to 1998). AB - INTRODUCTION: Group G streptococcus (GGS) accounted for 8% to 44% of all bacteraemias due to beta-haemolytic streptococci according to various reports. The aims of this study were 1) to describe the epidemiology of GGS bacteraemia in Singapore for which local data are lacking and 2) to compare its frequency of isolation to the other Lancefield groups. PATIENTS AND METHODS: The study period was from 1 January 1996 to 30 June 1998. The laboratory records of 2 large acute care hospitals were examined. There was a total of 85 patients. The medical records of 52 patients were available for analysis. In addition, laboratory microbiological data from 1993 to 1999 were reviewed and the number of blood cultures that were positive for beta-haemolytic streptococci groups A, B, C and G was collated. RESULTS: The majority involved the elderly. The mean age was 67 years. The skin was the major portal of entry. Local conditions predisposing the skin to infection occurred in 40.4%. Co-morbidity included malignancies in 28.8% of patients, diabetes mellitus in 11.5% and liver disease in 9.6%. Mortality was 15.4% including fatal septic shock. Recurrent bacteraemia occurred in 5.8% of the patients. The majority (90.4%) were community-acquired infections. GGS, along with group B streptococcus (GBS), was the most common streptococcus among the beta-haemolytic streptococci causing bacteraemia in these 2 hospitals. PMID- 11885505 TI - Bacteriologically-negative pulmonary tuberculosis--the Singapore tuberculosis control unit experience. AB - INTRODUCTION: Patients with radiological features suggestive of active pulmonary tuberculosis (PTB) from areas with a high prevalence of the disease have a high clinical likelihood of PTB regardless of the bacteriological findings. It is the established practice in many countries to initiate therapy in such patients. Our study aimed to determine if treatment for bacteriologically-negative PTB in our local population was appropriate and to identify features at presentation that would be predictive of active PTB, as defined by good and appropriate response to anti-tuberculous treatment. MATERIALS AND METHODS: A retrospective review of a randomised sample consisting of 100 bacteriologically-negative PTB patients given a course of anti-tuberculous treatment at the Singapore Tuberculosis Control Unit (TBCU). Based on their treatment response and outcome, patients were classified as probable active or unlikely active PTB. Patients' characteristics, clinical presentation and radiological findings were analysed for their association with likelihood of probable active PTB. RESULTS: Fifty-six per cent of patients in this study had probable active PTB. The decision to treat this group of patient was appropriate. There was no serious adverse reaction in the patients treated. The presence of symptoms, especially cough at presentation, a history of contact with tuberculosis and cavitation on chest radiograph, were associated with an increase risk of probable active disease. CONCLUSION: The TBCU's practice to treat patients suspected of having radiological PTB in the setting of negative sputum smear and culture seems to be appropriate in the majority of cases. PMID- 11885506 TI - Hepatolithiasis--a case series. AB - INTRODUCTION: Hepatolithiasis is an uncommon entity in Singapore. We reviewed the cases presented to our institution (a 1200-bedded restructured hospital) over a 5 year period. MATERIAL AND METHODS: Twelve cases of hepatolithiasis were treated between December 1995 and July 2000 representing 0.77% of gallstone disease operated on in the same period. The clinical presentation, investigations, treatment and subsequent progress are presented. RESULTS: The patients' ages ranged from 28 to 82 years. There was a male to female ratio of 5:7. All patients had upper abdominal pain at presentation; 10 patients had clinical evidence of cholangitis. Ultrasound was the commonest first line investigation but additional investigations, such as computed tomographic (CT) scan and various forms of cholangiography, were frequently necessary for complete delineation of the biliary disease. The intrahepatic stones were located predominantly in the left lobe of the liver. Parenchymal atrophy was seen in 83% of patients. Two patients underwent a biliary bypass operation only, 5 had a hepatic resection only, and 5 had combined procedures. Follow-up ranged from 4 to 50 months. Postoperative recovery was generally unremarkable. Complications included subphrenic abscess (1 patient), recurrent stricture (1 patient) and recurrent stones (1 patient). One patient had an elevated serum CA 19-9 preoperatively; a small villous adenoma was noted at the biliary stricture in the resected left lateral segment of the liver. There was no operative mortality. CONCLUSION: Hepatolithiasis is uncommon in Singapore. Complete diagnosis requires a combination of imaging modalities. Surgery remains the mainstay of definitive treatment. With adequate treatment, good outcome is possible. PMID- 11885507 TI - Plasma lipid peroxidation and vitamin E levels in smokers. PMID- 11885508 TI - Monitoring of presterile disposable syringes and needles marketed in Pune. AB - Out of hundred syringes and hundred needles (both presterile, disposable) tested, 41 syringes (41%) and 6 needles (6%) showed aerobic growth. Forty of the above syringes were tested simultaneously for fungi and anaerobes. Fungi isolated were 5.7% and no anaerobe was grown. Fifty glass syringes and fifty needles autoclaved in the departmental laboratory served as controls and did not show any growth. As a preventive measure, proper disposal of used disposable material should be made mandatory so that it does not find its way into the market. Or has the time come to switch back to the former conventional practice of using in house autoclaved articles? PMID- 11885509 TI - Protein profile in leprosy. AB - Serum proteins and plasma fibrinogen were estimated in 103 patients in various groups of leprosy and 52 patients of reactional leprosy. Total proteins, serum globulin and fibrinogen showed significant rise while serum albumin showed fall over the immunological spectrum from TT to LL. Type II reactional leprosy similarly revealed significant rise in globulin and fibrinogen. The comparison of these parameters between most of the comparable groups of leprosy was statistically significant. ENL patients after complete subsidence of reaction and after steroid treatment showed significant decrease in these protein fractions, thus conferring some prognostic implication on these tests. PMID- 11885510 TI - Role of percutaneous bone marrow grafting in delayed unions, non-unions and poor regenerates. AB - The present study comprises of 72 patients of post traumatic delayed unions, established non-unions, poor regenerate in segmental bone transportation and limb lengthening procedure treated by percutaneous injections of autogenous bone marrow at the site of failed healing. The average follow up was 4 years. Bone union was achieved in 68 patients. Overall, 72.2% of the patients had an excellent result, 11.1% a good result, 11.1% a fair result and 5.5% a poor result or failure. These results with only 4 failures (5.5%) are encouraging and suggest that percutaneous autogenous bone marrow grafting is a simple, safe and useful technique in the treatment of delayed unions and non-unions. We believe that this technique of percutaneous autogenous bone marrow grafting can be a procedure of choice in those patients where Phemister or Forbes methods of bone grafting alone is required, especially in limbs with scarred and poor soft tissue coverage. This procedure can also be useful in iatrogenic delayed or non-unions which is the commonest cause of non-union in present era of enthusiastic fracture fixation. PMID- 11885511 TI - Cutaneous nocardiosis in east Delhi--a case report. PMID- 11885512 TI - A crisis in morality. PMID- 11885513 TI - Root resection revisited. PMID- 11885514 TI - [Hereditary cerebral amyloid angiopathies]. AB - Cerebral amyloid angiopathies are defined by the presence of amyloid substance in the walls of cerebral vessels. All amyloid substances have a particular physico chemical structure, which imparts certain specific staining properties, but the biochemical composition of different amyloid types varies. Different forms of cerebral amyloid angiopathy have been identified, based on the biochemical nature of the protein deposited (e.g. beta-amyloid, cystatin C, transthyretin, gelsolin, amyloid protein Bri, prion protein). Some cerebral amyloid angiopathies are familial; these prompted genetic studies which in turn led to a better understanding of the genes coding for different amyloid proteins. As a group, cerebral amyloid angiopathies have certain neuropathological lesions in common. Infiltration by amyloid substance results in weakening of the small vessel walls and secondary complications responsible for changes such as microinfarcts and miliary haemorrhages in the cerebral cortex, lobar haemorrhages and/or leucoencephalopathy. These changes form the basis of the neurological complications: meningeal and cerebral haemorrhages, transient ischaemic episodes, vascular dementia. However each type of hereditary cerebral amyloid angiopathy has individual clinical and histopathological features reflecting the severity of arterial involvement, the extent of amyloid deposition within or outside the central nervous system, and the association with other neurodegenarative changes. PMID- 11885515 TI - [Headaches caused by abuse of symptomatic anti-migraine and analgesic treatment]. AB - These daily or near-daily headaches result from the chronic overuse of all immediate relief antimigraine drugs: ergotamine, analgesics, and/or more recently triptans. Like for much chronic daily headaches, the International Headache Society diagnostic criteria for drug abuse headaches are difficult to apply. Generally, patients confuse primary headaches (usually migraines) with interparoxysmal tension-type headaches called "rebound headaches". Psychosocial factors may play a role. Insidiously, a compulsive automedication results, often in anticipation of headache. This headache syndrome resists symptomatic and prophylactic treatment. These headaches are frequent, very disabling and socioeconomically costly. They are still largely underdiagnosed. Drug-induced headaches may be restricted to those patients who are already headache sufferers. The pathogenesis is not clearly understood: it may involve a deficience of inhibitory pain modulation, a hyperactivation of nociceptive facilitatory systems, and the peripheral and central effects of the incriminating drugs. The withdrawal of all offending analgesic drugs and a multimodality approach are indispensable, but the therapeutic protocoles are actually very heterogeneous and poorly estimated. Non-drug means could be very helpful. Effective education of headache sufferers and regular follow-up are essential to avoid relapses. Prognosis factors have been evoked, but may not be significant for the long term outcome. The rate successfull of is actually estimated at 60 p. cent at five years. The benefits of an adequate management encourage early recognation of drug induced headaches. This article has in view to take stock of the literature at the end of 1999, and to help physicians become mora aware of this problem and develp a more preventive attitude. PMID- 11885516 TI - [Illusions of body normality in amputees and paraplegic patients]. AB - Phantom limbs in amputees, or body illusion in hemiplegics, have been the subject of wide ranging descriptions. The detected abnormalities involve morphological, postural and/or kinetic features. The aim of this prospective study carried out in 25 amputees and 10 adult paraplegics was to describe the typology of these perceptions. Data were collected from free and semi-directive investigations before and after caloric vestibular stimulation. Amputees and paraplegics perceived normal, deformed and painful body phantom segments, reffered perceptions and "normal limbs" which took on the request posture considering the general body position (illusion of body normality). This perception corresponds to an image of the body, such as it should be and not such as it is. In amputees, the limb follows the movements of the prothesis. These perceptions conform quite well reality so that the loss of the paralyzed limb is not perceived as a missing limb. This illusion of body normality should be distinguished from the normal phantom limb, characterized by a stronger perception of the lost limb compared with the other. In both amputees and paraplegics, vestibular stimulation can generate or modify phantoms limbs or body illusion and can abolish painful phantom limbs. The neuromatrix, which rebuilds body representations, could get its information from reorganized cortical areas (instantaneous body image), autobiographical engrams (painful phantoms limbs), or innate engrams (identity body schema) that, via congruence mechanisms, could be identified as a somatic reference, particularly for motor programming. This interpretation is compatible with current knowledge and suggests how amputees can easily use a prothesis. PMID- 11885517 TI - [Conduction aphasia and phonemic disorder]. AB - Conduction aphasia is usually described as a repetition impairment. Semiology or pathophysiology cannot be explained with this definition. We report a single case particularly demonstrative. The patient showed spontaneous speech, denomination, repetition and reading impairments. Main errors were phonemic paraphasia. No arthric disorder nor comprehension impairment was observed. Damage of supramarginalis gyrus and Wernicke's area was found. A cognitive analysis suggested that the phonological buffer and the working memory were impaired. Implication for rehabilitation, which included segmentation and semantisation associated to phonological training, is discussed. The course of the conduction aphasia was good and the patient was able to work again. PMID- 11885518 TI - [Ischemic cerebrovascular stroke of arterial origin in the child]. AB - Comparing stroke in children with stroke in adults can provide interesting information because age and cerebral plasticity induce specific clinical features and outcome. Arterial ischemic strokes are the most frequent in childhood although the problem is not one of arteriosclerosis. Arterial dissection, Moya Moya syndrome, and cardioembolic and thrombogenic events induced by hemoglobin diseases and hyperhomocyteinemia must be detected at the first event. In some cases, onset is marked by head trauma or an infectious syndrome. The important feature is that outcome is better than in adults and is marked by onset of hemidystonia, partial epilepsy. Aphasia is benign if stroke arises before the child has learned to write. PMID- 11885519 TI - [Multiple intracranial and intraspinal meningiomas successively discovered in the absence of neurofibromatosis: 2 cases]. AB - Multiple meningiomas in different neuroaxial compartments are quite rare. We describe the case of a 44-year-old woman who developed three intracranial meningiomas and 8 years later a T3 dorsal meningioma. Histologically, the frontal and dorsal tumors appeared as benign psammomatouss meningiomas. Both tumors were removed successfully. The second patient was a 31-year-old woman who developed right benign fronto-parietal transitional meningioma. She presented local and spheno-orbital recurrences, then a lombo-sacral lesion. The histological picture worsened from benign to malignant with multiple recurrences. Several mechanisms could account for multiple meningiomas. Such meningiomas could arise from a single primary tumor via subarachnoidal spread of a benign or malignant nature. Alternatively, they could be atypical forms of neurofibromatosis type 2 or tumors with a multifocal origin. PMID- 11885520 TI - [Diurnal consequence of insomnia: impact on quality of life]. AB - Insomnia is not only a disease of sleep, it has also daily consequences: fatigue, irritability, impaired daytime functioning. These complaints are regent reported by the patients, however the objective tests assessing alertness in insomnia are usually not impaired when compared with good sleepers. We wanted to appreciate more accurately the daily consequences of insomnia, in terms of quality of life. 240 severe insomniacs (according to the DSM-IV criterias) and 391 good sleepers received a questionnaire on quality of life items. Depressed and anxious patients were excluded from this group. The questionnaire was built by a multidisciplinary group, based on insomniac's interviews. It was primarily tested in a small sample and then proposed in the entire group. Insomniac's quality of life appeared to be significantly impaired in comparison with good sleepers. They experienced more fatigue and more sleepiness during the daytime. They reported more attention disorders and memory complaints. They seemed to be more irritable and sensitive to the environment. At work they made more mistakes and had more sic leave. They also had poorer relationships with relatives and family than good sleepers. PMID- 11885521 TI - [Andermann syndrome in an Algerian family: suggestion of phenotype and genetic homogeneity]. AB - Andermann syndrome or Agenesis of the Corpus Callosum with Polyneuropathy (MIM 218000) is an autosomal recessive disease almost exclusively found in Quebec. Only few cases have been reported in other populations. The locus for Andermann syndrome was assigned to chromosome 15q13-q15 in French Canadian families. We performed a haplotype analysis with two markers of this chromosomal region in an Algerian consanguineous family with two affected sibs. The children were homozygous for both markers, suggesting genetic homogeneity in Andermann syndrome. PMID- 11885522 TI - [Primary hypereosinophilia syndrome manifesting as encephalopathy and vision disorders]. AB - We report the case of a 60-year-old man who developed visual and cognitive disorders. Investigations confirmed the diagnosis of idiopathic hypereosinophilic syndrome even though the patient had a history of rectal cancer. The olinical course was favorable after treatment. We discuss the different clinical forms, imaging data and treatments of eosinphilic syndrome. PMID- 11885523 TI - [Hemichorea caused by striatal infarct in a young type 1 diabetic patient]. AB - Movement disorders such as chorea and ballism rarely occur in diabetes mellitus. We report the case of 26-year-old man with a 13-year-history of type 1 diabetes mellitus. He presented with a right side hemichorea. Brain CT-scan and MRI showed an infarction of the head of the caudate nucleus and the anterior part of the putamen. Presence of microangiopathy affecting retina, kidneys and peripheral nerves suggest a similar involvement of the lenticulo-striatal arteries. Hemichorea and hemiballism usually occur in older patients presenting type 2 diabetes mellitus. Non-ketotic hyperglycaemia is the common cause in such situation. Striatal infarct, as seen in our patient, is rarely reported. PMID- 11885524 TI - [Multiple mononeuropathy and inflammatory syndrome manifested in Lyme disease]. AB - Meningo-radiculitis is the most common peripheral nerve system involvement of Lyme disease. We report the observation of a 73 year-old woman presenting a subacute multiple mononeuropathy and a severe inflammatory syndrome. Diagnosis of Lyme disease was confirmed by a lymphocytic meningitis with positive serologic results in the cerebrospinal fluid. Nerve biopsy showed inflammatory cells spreading along the endoneurium. This case report emphasizes that Lyme disease may present as a multiple mononeuropathy mimicking a vasculitic neuropathy. PMID- 11885525 TI - [Tubular aggregate congenital myopathy associated with neuromuscular block]. AB - Various myopathies are described associated with tubular aggregates. However, in several cases tubular aggregates constitute the main structural feature allowing to consider myopathy with tubular aggregates as a distinct entity. A 50-year-old woman whose parents were consanguinous, presented frequent falls. She walked only after 18 months of age and did poorly in gymnastics. The weakness, which has myasthenic feature, involved predominantly the pelvis girdle. The serum creatine kinase was 206 UI/L (normal < 110 UI/L). Electromyogram showed a myogenic pattern in proximal muscles. Repetitive stimulation on the trapezius revealed 50 p. cent decrementing response. Muscle biopsy showed numerous tubular aggregates in type II fibers. Anti-acetylcholine receptor (AChR) antibodies were absent. There was no thymoma. The neostigmine test was negative. Clinical and electrical myasthenic features characterize one of the numerous forms of myopathy with tubular aggregates. In our case, the lack of AChR antibodies and the negative response to neostigmine argue in favor of a dysfunction of the AChR. This unusual observation highlights the therapeutic difficulties in this myopathy with neuromuscular block. PMID- 11885526 TI - [Is it necessary to treat persistent aneurysms after dissection of the cervical arteries?]. PMID- 11885527 TI - [Ophthalmologic and neuro-ophthalmologic examination of the patient with multiple sclerosis]. PMID- 11885528 TI - [Abnormal diurnal somnolence, "sleep attacks" and antiparkinson drugs]. PMID- 11885530 TI - Use of gamma radiation for treatment of renal artery in-stent restenosis. PMID- 11885529 TI - [To the memory of professor Lev Polezhaev]. PMID- 11885531 TI - [Microbiological diagnostics in periodontal treatment]. AB - Periodontal diseases are bacterial infections. The rationale for the use of microbiological diagnostics in periodontal treatment of severe periodontitis patients is discussed as well as the use of adjunct antimicrobial therapy for the elimination of specific bacterial species. PMID- 11885532 TI - [Bleeding tendency and periodontal diagnosis. Evaluation of the prognostic significance of bleeding tendency for periodontal diagnosis]. AB - Since differences in susceptibility to periodontal breakdown do exist, it is essential to predict future breakdown on the basis of a prognostic indicator. The experimental gingivitis model was chosen to investigate the clinical differences related to the degree of susceptibility to periodontal breakdown. Both bleeding upon probing and the amount of plaque seems to depend on the history of inflammatory periodontal disease. It is concluded that the bleeding/plaque ratio may act as a prognostic indicator to predict periodontal breakdown. PMID- 11885533 TI - [Periodontal inflammation. Biochemical markers in crevicular fluid for diagnosis of periodontal disease]. AB - Based on a review of the literature the possible application of biochemical markers in the early detection of periodontal disease is discussed, using crevicular fluid. PMID- 11885534 TI - [Patient care and dental ethics. The request of a patient for total extraction]. AB - On the basis of a case report in which a patient requests total extraction while, from the view of dental care, there are possibilities to preserve the teeth, the question is discussed if there is a moral justification to gratify the wishes of the patient. PMID- 11885535 TI - Three years postgraduate programme in orthodontics. Final report Erasmus project. AB - A new curriculum with a common contents of 75%, leaving 25% for electives has been developed for a three year programme for specialty education in orthodontics. The result is a joint effort of the original eight participants and invited professors from Italy and Ireland as well as from five non-EEC countries (Austria, Finland, Norway, Sweden and Switzerland). Unanimity among the 15 participants was reached on all essential matters and on the contents of the final report. The main objective of the programme has been defined and the different courses have been formulated in terms of goals to be accomplished in three levels of comprehension and by the minimal hours students must devote to each subject. Furthermore, procedures that should be mastered have been specified. PMID- 11885536 TI - [Diagnosis and treatment planning of the class II/2 malocclusion]. AB - Diagnosis and treatment planning of the Angle class II/2 malocclusion of the dentition are discussed. The skeletal pattern is essentially normal except for an anterior rotation of the mandible during growth. Major deviations involve the dentition. The high lip line is of paramount importance for the development of the Class II/2 malocclusion. PMID- 11885537 TI - [Patient care and dental ethics. III. The advice of an acupuncturist for the removal of dental amalgam]. AB - On the advice of an acupuncturist a patient requests her dentist to remove her amalgam fillings which are of a good quality. The question is discussed whether there is a moral justification to gratify the wishes of the patient. PMID- 11885538 TI - [Chronic facial pain due to nerve injury. Deafferentation pain]. AB - Patients presenting chronic facial pain are very common. After profound examination a significant cause for this pain often cannot be found. Some patients are suffering from this pain due to injury of a sensible nerve and then it is called deafferentation pain. This somatic disorder can give different complaints. Pain can develop as a result of formation of neuromas 'cross-talking' of nerves and alterations in the central nerve system. It seems to be difficult to make clinical features to diagnose deafferentation pain and differentiate it from pain disorders where the psychic suffering seems to be very important. The treatment is based mainly on the reduction of the conducting of abnormal pulses by the damaged or changed nerves. PMID- 11885539 TI - [When the heart beats faster: negative expectations about dental treatment]. AB - Many people expect dental treatment to be painful. This kind of expectation has important negative consequences for the patients' well-being. Disappointing occurrences during the treatment itself can further influence the expectations about a next treatment. The fact that the treatment result happens to be more positive than expected is a necessary instead of a sufficient condition for changing existing expectations. A cognitive model for dental fear is proposed and some implications for the general practice are discussed. PMID- 11885540 TI - [Dental anxiety]. AB - The variety of patients with dental anxiety is large. A classification is made containing four types of patients, using criteria related to fear complexity and specificity. Analysing the issues provoking dental fear, the anxiety for loss of control is a major one. Procedures of guiding and treating patients related to their classification have been briefly described. PMID- 11885541 TI - [Patient care and ethics. The dentist in love with his patient]. AB - A young dentist, who just started his practice, falls in love with a 24-year old woman, during an expensive treatment of several crowns and bridges. He gets confused if it is wise for him to continue this treatment. This question is discussed by a philosophical analysis of what is called the 'I-am-in-love situation'. PMID- 11885542 TI - [Darkfield or phase contrast microscopy. Usefulness in periodontology]. AB - Microscopic evaluation of a dental plaque sample is not very useful, since the bacteria are difficult to distinguish from the diluent (same refractive index). Two types of microscopic analyses try to solve this problem in a different way. Using a darkfield microscope, the object is illuminated by slanting rays of light, that are then dispersed or bent away and enter the object. In this way, a shining image on a dark background is formed. The phase contrast microscope uses two principles of the geometry (wave length and amplitude) to create an image of the illuminated cells. Methodologically the next aspects are important, since they strongly influence the outcome of the analysis: contamination of the sample, technique of sampling and preparation of the sample. The reproducibility of the above mentioned techniques is high when a great number of parameters is kept constant. The analysis of the sample give us some clinically relevant information. PMID- 11885543 TI - Neuromuscular disorders: gene location. PMID- 11885544 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 11885545 TI - A conservative alternative. PMID- 11885546 TI - Readers' perspectives. Most health care CIOs have adequate disaster recovery plans. PMID- 11885547 TI - [Acute renal insufficiency. Diagnostic algorithm]. PMID- 11885548 TI - Different quantities and quality of fat in milk products given to young children: effects on long chain polyunsaturated fatty acids and trans fatty acids in plasma. AB - In this study we compared plasma contents of long-chain polyunsaturated fatty acids (LC-PUFAs) and trans fatty acids in triglycerides (TG), phospholipids (PL) and cholesterolesters (CE) in young children fed milk diets containing different amounts of linoleic (LA) and alpha-linolenic acid (ALA). Because the diets differed in vitamin A and E content, plasma concentrations of vitamin A and E were also studied. Thirty-seven 1-y-old children were randomly assigned to one of four feeding groups: (1) low-fat milk (LF) (1.0 g cow's milk fat/dL); (2) standard-fat milk (SF) (3.5 g cow's milk fat/dL); (3) partially vegetable fat milk (PVF) (3.5 g fat/dL; 50% vegetable fat from rapeseed oil, 50% milk fat); and (4) full vegetable fat milk (FVF) (3.5 g fat/dL; 100% vegetable fat from palm-, coconut- and soybean oil). We found higher amounts of plasma LA in the FVF group than in the LF and SF groups (p < 0.001) and higher amounts of ALA in the PVF group than in the SF (p < 0.001 in TGs, p < 0.05 in CEs) and LF (p < 0.01 in PLs and CEs, p < 0.05 in TGs) groups. However, amounts of plasma arachidonic acid (AA) were similar between groups as well as the amounts of docosahexaenoic acid (DHA) in CEs and PLs. Total trans FAs were lower in CEs in the PVF and FVF groups than in the SF group (p < 0.05 SF vs PVF; p < 0.01 SF vs FVF). Plasma concentrations of alpha-tocopherol were higher in the FVF group than in the other groups (p < 0.05 FVF vs SF, p < 0.01 FVF vs SF and PVF). CONCLUSION: Children consuming milk diets containing high amounts of vegetable fat present with higher plasma LA and ALA without any effects on amounts of plasma LC-PUFA. The plasma LC PUFA status is not adversely affected by a low-fat milk diet. AHA and DHA in plasma are not affected by the diets studied, presumably because 15-mo-old children may be able to compensate for dietary influences through endogenous LC PUFA metabolism. PMID- 11885549 TI - Laboratory-confirmed reinfections with Bordetella pertussis. AB - Susceptibility to infection with Bordetella pertussis re-emerges several years after pertussis vaccination. However, the duration of immunity after natural infection with B. pertussis, postulated to be lifelong, is not known. In an ongoing study, the longitudinal course of pertussis antibodies in patients who have had laboratory-confirmed pertussis is being followed using sera obtained at irregular intervals. In 4 patients a reinfection with Bordetella pertussis is described respectively 7 (patient A), 12 (patients B and C) and 3.5 (patient D) y after the first infection. It seems that the longer the interval between the infections the more severe the complaints. CONCLUSION: To the authors' knowledge. these are the first patients in whom symptomatic reinfection with B. pertussis has definitely been proven by laboratory confirmation of both episodes. Bordetella pertussis infection should be considered in patients with symptoms of typical or atypical whooping cough, irrespective of their vaccination status or previous whooping cough. PMID- 11885551 TI - Peripheral disorders in the otolith system. A pathophysiological and clinical overview. PMID- 11885550 TI - The mammalian otolithic receptors: a complex morphological and biochemical organization. PMID- 11885552 TI - Pathophysiology and clinical testing of otolith dysfunction. PMID- 11885553 TI - Otolithic vertigo. PMID- 11885554 TI - Physiopathology of otolith-dependent vertigo. Contribution of the cerebral cortex and consequences of cranio-facial asymmetries. PMID- 11885555 TI - Clinical and instrumental investigational otolith function. PMID- 11885556 TI - The subjective visual vertical. PMID- 11885557 TI - Clinical application of the off vertical axis rotation test (OVAR). PMID- 11885558 TI - VEMP induced by high level clicks. A new test of saccular otolith function. PMID- 11885559 TI - Total cortisol levels are reduced in the periovulatory follicle of infertile women with minimal-mild endometriosis. AB - PROBLEM: To measure and compare concentrations of total and free glucocorticoids with oocyte fertilizing capacity in the follicular fluid (FF) of women with minimal-mild endometriosis and tubal damage. METHOD OF STUDY: Follicular fluid was collected from individual periovulatory follicles during oocyte retrieval for in vitro fertilization (IVF) in natural cycles. Total and free levels of cortisol and cortisone were measured using specific radioimmunoassays after chloroform extraction. RESULTS: Cortisol concentrations in women with minimal-mild endometriosis were significantly lower compared with controls (women with tubal infective damage) (258 versus 328 nmol/L, P < 0.02). There was no correlation between total or free concentrations of cortisol or cortisone and the fertilization capacity of the oocyte. CONCLUSIONS: Total cortisol levels are lower in the follicles of women with endometriosis. Our findings provide further evidence of follicular dysfunction contributing to the subfertility associated with minimal-mild endometriosis. PMID- 11885560 TI - Experimental and computational screening models for prediction of aqueous drug solubility. AB - PURPOSE: To devise experimental and computational models to predict aqueous drug solubility. METHODS: A simple and reliable modification of the shake flask method to a small-scale format was devised, and the intrinsic solubilities of 17 structurally diverse drugs were determined. The experimental solubility data were used to investigate the accuracy of commonly used theoretical and semiexperimental models for prediction of aqueous drug solubility. Computational models for prediction of intrinsic solubility, based on lipophilicity and molecular surface areas, were developed. RESULTS: The intrinsic solubilities ranged from 0.7 ng/mL to 6.0 mg/ mL, covering a range of almost seven log10 units, and the values determined with the new small-scale shake flask method agreed well with published solubility data. Solubility data computed with established theoretical models agreed poorly with the experimentally determined solubilities, but the correlations improved when experimentally determined melting points were included in the models. A new, fast computational model based on lipophilicity and partitioned molecular surface areas, which predicted intrinsic drug solubility with a good accuracy (R2 of 0.91 and RMSEtr of 0.61) was devised. CONCLUSIONS: A small-scale shake flask method for determination of intrinsic drug solubility was developed, and a promising alternative computational model for the theoretical prediction of aqueous drug solubility was proposed. PMID- 11885561 TI - Relation between K+ leakage and damage to band 3 in photodynamically treated red cells. AB - Potassium leakage is one of the first events that appear after photosensitization of red blood cells. This event may subsequently lead to colloid osmotic hemolysis. The aim of our study was to determine which photodynamically induced damage is responsible for increased membrane cation permeability. This was done by studying the effect of dimethylmethylene blue (DMMB)-mediated photodynamic treatment (PDT) on different membrane transport systems. Inhibition of band 3 activity (anion transport) showed a comparable light dose dependency as PDT induced potassium leakage, whereas glycerol transport activity was inhibited only at higher light doses. Dipyridamole (DIP), an inhibitor of anion transport, protects band 3 against DMMB-induced damage, and prevents the increase in cation permeability of the membrane. Damage to glycerol transport was partially reduced when PDT was performed in the presence of DIP. Because DIP has no affinity for the glycerol transporter, this protection might result from the reduced photodamage to band 3. These results support the hypothesis that band 3 might be involved in glycerol transport. Glucose transport was not affected by DMMB mediated PDT. The present results are the first to show a causal relationship between DMMB-mediated photodamage to band 3 and increased cation permeability of red blood cells. PMID- 11885562 TI - Systemic component of protoporphyrin IX production in nude mouse skin upon topical application of aminolevulinic acid depends on the application conditions. AB - Topical application of 5-aminolevulinic acid (ALA) for protoporphyrin IX (PpIX) based photodynamic therapy of skin cancer is generally considered not to induce systemic side effects because PpIX is supposed to be formed locally. However, earlier studies with topically applied ALA have revealed that in mice PpIX is not only produced in the application area but also in other organs including skin outside the application area, whereas esterified ALA does not. From these results, it was concluded that it is not redistribution of circulating PpIX that causes the fluorescence distant from the ALA application site, but rather, local PpIX production induced by circulating ALA. In the present study we investigate the effects of the ALA concentration in the cream, the application time, the presence of a penetration enhancer, the presence of the stratum corneum and esterification of ALA on the PpIX production in nude mouse skin outside the area where ALA is applied. For this purpose, ALA and ALA hexyl ester (ALAHE) were applied to one flank, and the PpIX fluorescence was measured in the contralateral flank. During a 24 h application of ALA, PpIX was produced in the contralateral flank. No PpIX could be detected in the contralateral flank after ALA application times ranging from 1 to 60 min. Tape-stripping the skin prior to short-term ALA application, but not the addition of a penetration enhancer, resulted in PpIX production in the contralateral flank. When ALAHE was applied, no PpIX fluorescence was measured in the contralateral flank under any application condition. The results suggest that the systemic component of PpIX production outside the ALA application area plays a minor or no role in relevant clinical situations, when the duration of ALA (ester) application is relatively short and a penetration enhancer is possibly added. PMID- 11885563 TI - The myc oncogene: MarvelouslY Complex. AB - The activated product of the myc oncogene deregulates both cell growth and death check points and, in a permissive environment, rapidly accelerates the affected clone through the carcinogenic process. Advances in understanding the molecular mechanism of Myc action are highlighted in this review. With the revolutionary developments in molecular diagnostic technology, we have witnessed an unprecedented advance in detecting activated myc in its deregulated, oncogenic form in primary human cancers. These improvements provide new opportunities to appreciate the tumor subtypes harboring deregulated Myc expression, to identify the essential cooperating lesions, and to realize the therapeutic potential of targeting Myc. Knowledge of both the breadth and depth of the numerous biological activities controlled by Myc has also been an area of progress. Myc is a multifunctional protein that can regulate cell cycle, cell growth, differentiation, apoptosis, transformation, genomic instability, and angiogenesis. New insights into Myc's role in regulating these diverse activities are discussed. In addition, breakthroughs in understanding Myc as a regulator of gene transcription have revealed multiple mechanisms of Myc activation and repression of target genes. Moreover, the number of reported Myc regulated genes has expanded in the past few years, inspiring a need to focus on classifying and segregating bona fide targets. Finally, the identity of Myc-binding proteins has been difficult, yet has exploded in the past few years with a plethora of novel interactors. Their characterization and potential impact on Myc function are discussed. The rapidity and magnitude of recent progress in the Myc field strongly suggests that this marvelously complex molecule will soon be unmasked. PMID- 11885564 TI - [The dispute over eugenics in interwar Poland]. AB - The eugenic problem seems not as well known as it was in the period between the world wars. The word "eugenics" was introduced into the scientific language by the British biologist, Francis Galton at the end of the XIX century. In general, it means the study of the methods of protecting and improving the quality of the human race by selective breeding. Galton's remarks initiated a wider (social and political) movement, which was active in some countries. The eugenic tendency was visible in American immigration policy before the Great War - making it difficult for people suffering from particular diseases to enter the USA. After the war, the eugenic movement became much stronger. In some countries (e.g. Sweden, Norway, Finland, Denmark, Switzerland, Germany, and over half of the states of the USA) some components of the eugenic programme were introduced into legislation. The eugenic movement also appeared in reconstructed Poland. In 1918 it published it own magazine "Zagadnienia Rasy" (Problems of Human Race), later "Eugenika Polska" (Polish Eugenics). This did not imply however, that interest in the eugenic programme was generally very strong. In fact, it was limited by the influence of the Catholic Church. Another significant factor was lost Polish - German hostility. In the late thirties eugenic slogans lost popularity because of their use in Germany racist policy. PMID- 11885566 TI - Chlorinated pesticides in mussels from Guanabara Bay, Rio de Janeiro, Brazil. PMID- 11885565 TI - Current issues in dendritic cell cancer immunotherapy. AB - Dendritic cells (DCs) initiate and direct the immune response. Their inability to detect danger signals from transformed cells and to generate an effective immunological response may allow cells with a malignant phenotype to evolve into cancers. This defect can be corrected for many cancer types and the immune response boosted to eliminate malignant cells by means of DC-based vaccines/therapies. Rapid advances in our understanding of basic DC physiology and improved methods for DC isolation have made clinical application of DC therapy practical, and encouraging phase I/II results are emerging. PMID- 11885567 TI - Serotonergic and cognitive impairment in impulsive aggressive personality disordered offenders: are there implications for treatment? AB - BACKGROUND: Reduced serotonin (5-HT) function and deficits on neuropsychological tasks have been separately reported in antisocial populations. We investigated whether these impairments are independent or associated factors underlying impulsivity in aggressive personality disordered (PD) offenders and healthy controls and whether there are associated changes in quantitative brain measures. METHODS: This study reports on the findings from a sample of 51 PD offenders and 24 controls, recruited from maximum security psychiatric hospitals, who were characterized using the Special Hospital Assessment of Personality and Socialisation (SHAPS). Subjects underwent assessment of 5-HT function (prolactin response to D-fenfluramine challenge), neuropsychological testing and had a diagnostic MRI scan. Of this sample 19 controls and 24 patients also had quantitative measurement of frontal and temporal lobe volumes on magnetic resonance imaging (MRI). RESULTS: Non-psychopathic (low-impulsive) aggressive PDs had enhanced 5-HT function compared with controls and highly impulsive aggressive psychopaths. Primary and secondary psychopaths had poorer executive/frontal, but not memory/temporal neuropsychological function than controls and non psychopaths. There were no significant group differences in frontal or temporal lobe brain volumes. Although impulsivity and aggression are correlated constructs impulsivity appeared to be related to both executive function and 5-HT function, while aggression only correlated inversely with executive/frontal and memory/temporal function. 5-HT did not directly correlate with frontal or temporal volume or function. CONCLUSION: Impulsivity appears to be contributed to by both impaired neuropsychological function and 5-HT function. Impaired neuropsychological function alone makes a contribution to aggression. Treatment needs to take account of the neuropsychological and biochemical deficits in this challenging population. PMID- 11885568 TI - Acculturation and suicide: a case-control psychological autopsy study. AB - BACKGROUND: The relationships between acculturation and suicide were investigated in East Taiwan. METHODS: Psychological autopsy interviews were conducted for consecutive suicides from two native Taiwanese groups (Atayal and Ami) (N = 30 for each group); each of them was matched with two controls for age, sex and area of residence. The Taiwan Aboriginal Acculturation Scale was used to measure the extent of acculturation. RESULTS: A lower degree of social assimilation was significantly associated with a higher risk of suicide in the Atayal and the male groups. In multivariable regression analysis, a significant effect of low social assimilation on the risk of suicide was found in Atayal and in men, even after controlling for the effects of ICD-10 depressive episode and emotionally unstable personality disorder. Meanwhile, there was a significant trend across low, moderate and high social assimilation on suicide risk in Atayal and in men. CONCLUSIONS: For the native Taiwanese, the stress from rapid acculturation into the main Chinese society is crucial to their mental health. It might be reduced through targeted social and educational programmes. PMID- 11885569 TI - Mood congruent memory bias induced by tryptophan depletion. AB - BACKGROUND: Mood congruent memory bias predicts a more superior recall memory of learnt material congruent with the mood state at the time of learning. The present study is the first report of an experimental study in which a biological mood induction was used to test this hypothesis. The influence of acute tryptophan (TRP) depletion, inducing low serotonin neurotransmission and a depression of mood, on memory bias was evaluated in healthy volunteers (16 with and 11 without a family history of major affective disorder). METHODS: Twenty seven subjects received 100 g of an amino acid mixture with and without TRP according to a placebo-controlled, double-blind, balanced, cross-over design. An affective memory test consisting of a 30-word list with words of positive, neutral, and negative affective valence and a mood questionnaire were assessed at 6 and 24 h following treatment administration. RESULTS: TRP depletion impaired delayed recall of neutral and positive words, but not of negative words. There was no interaction of family history and treatment and there was no post hoc association between the influence of TRP-depletion on mood and on affective memory bias. CONCLUSION: Experimentally induced serotonergic depletion in normal individuals shifts affective memory bias towards negative affective valent verbal stimuli. PMID- 11885570 TI - Backbone and side-chain 1H, 15N, and 13C assignments for chick cofilin. PMID- 11885571 TI - Conversion from the SPEAK to the ACE strategy in children using the nucleus 24 cochlear implant system: speech perception and speech production outcomes. AB - OBJECTIVE: The main objective of this study was to assess whether speech perception and speech production in children using the Nucleus 24 cochlear implant system improved with a change in speech processing strategy from the SPEAK to the Advanced Combination Encoder (ACE) strategy. The major difference between the two strategies is that ACE uses a higher stimulation rate (in this study the stimulation rate was 900 Hz per channel) compared with the SPEAK strategy, where the stimulation rate is 250 Hz per channel. Information also was obtained regarding the adjustment period after conversion to the ACE strategy. DESIGN: An ABA experimental design was used where scores were initially obtained using the SPEAK strategy' (in the initial A time interval), and subsequently performance was assessed using the ACE strategy (B time interval) and then again with the SPEAK strategy (second A time interval). The duration of the B interval was 10 wk, and the duration for the second A interval was 4 wk. Seven children aged between 9 and 16 yr who had at least 6 mo experience with the SPEAK strategy participated. Open-set monosyllabic CNC word perception in quiet and Speech Intelligibility Test sentence perception in noise was evaluated at the end of each of the time intervals. Word perception was also monitored at fortnightly intervals during the B time interval. Speech production was assessed at the end of the initial A time interval and at the end of the B time interval. RESULTS: Mean word and phoneme scores for open-set words in quiet for the group of seven children were significantly higher with the ACE strategy as compared with the SPEAK strategy scores obtained in both of the A time intervals. For sentences in noise, mean scores using the ACE strategy as well as the SPEAK strategy at the second A evaluation point were significantly higher than the scores using the SPEAK strategy measured at the first A time interval. This suggests that learning effects may have influenced outcomes. For some subjects, an initial decrease in scores was found during the initial 2-wk period after fitting the ACE strategy; however, scores subsequently were found to be similar to or higher than those when using the initial SPEAK strategy. Analysis of speech production assessments showed an improvement in the medial consonant scores after using the ACE strategy. CONCLUSIONS: This study demonstrated that some children were able to benefit from the additional information provided by the ACE strategy as compared with the SPEAK strategy. However, the differences in overall performance between the two strategies appear to be relatively small. PMID- 11885572 TI - Determination of the release of hydrolyzed demethylcantharidin from novel traditional chinese medicine-platinum compounds with anticancer activity by gas chromatography. AB - The present paper describes the development of a simple, accurate and reproducible gas chromatographic method for the determination of hydrolyzed demethylcantharidin release from a novel series of traditional Chinese medicine (TCM)-platinum compounds possessing potent anticancer and protein phosphatase 2A (PP2A)-inhibition properties. The salient features of the validated assay were a limit of detection (LOD) of 2 microg/mL, a limit of quantitation (LOQ) of 6 microg/mL, an intra- and inter-day precision of less than 11%, and an accuracy of more than 92%. The developed GC-flame ionization detection (FID) method was successfully utilized for the analysis of hydrolyzed demethylcantharidin, the TCM component that is slowly released from the novel compounds over 24 h, leading to PP2A inhibition. Further structural confirmation was achieved by GC-MS. The GC method is suitable for further mechanistic, pharmacokinetic and metabolic studies of the TCM-Pt compounds that might prove to be new anticancer agents with novel mechanisms of cytotoxic action. PMID- 11885573 TI - A process risk model for the shelf life of Atlantic salmon fillets. AB - The shelf life of Atlantic salmon (Salmo salar) portions produced for retail distribution is examined and the dominant aerobic spoilage organism is identified. Characterization of the harvesting and processing operations allow the development of a stochastic mathematical model, a process risk model (PRM), which predicts the range of the possible shelf life for the portions under normal retail and distribution. The considered risk is the failure to achieve the nominal 'use by' date. Bacterial counts from surface swabs, water, ice, and fish samples, collected over a period of 9 months, are fitted to distribution functions for use within the model. Comparisons are made between the distributions fitted to the observed bacterial levels and the predicted levels for the slurry water, initial surface contamination on the fish, and for the predicted and observed shelf life. Storage temperature of the packaged salmon portions has the greatest influence on shelf life, with contamination from contact surfaces and other sources being the next most important. The range of bacterial counts on the portions was between -0.6 and 5 log10 cfu/cm2. The model predicts bacterial counts in the slurry water to have an average value of 3.36 log10 cfu/ml, whereas the observed slurry water bacterial counts were 3.35 log10 cfu/ml. The predicted average initial bacterial contamination is 3.31 log10 cfu/cm2 on the fish surface and 3.23 log10 cfu/cm2 on the observed. The average predicted shelf life is 6.5 days, compared to an observed value of 6.2 days at 4 degrees C. PMID- 11885574 TI - Antagonistic activity of Lactobacillus casei strain shirota against gastrointestinal Listeria monocytogenes infection in rats. AB - In the present study, the effect of ingested viable Lactobacillus casei Shirota strain YIT9029 on oral infection with the enteric pathogen Listeria monocytogenes in Wistar rats was investigated. Rats were orally infected with 10(9) viable L. monocytogenes. Starting 3 days before the infection, rats received a daily dosage of 10(9) viable L. casei. It was shown that supplementation of L. casei significantly reduced the numbers of L. monocytogenes in stomach, caecum, faeces, spleen and liver, 2 days after L. monocytogenes infection. The number of L. monocytogenes in the mesenteric lymph nodes was not affected by the ingestion of L. casei. In comparison with control animals, the levels of the liver-specific alanine aminotransferase were lower in L. casei-fed rats. Histological analysis of spleen and liver revealed no differences between the experimental and control animals. In a parallel study with orally L. monocytogenes infected rats, it was shown that L. casei was able to increase cellular immunity significantly as determined with the delayed-type hypersensitivity response against heat-killed L. monocytogenes. In conclusion, in the present study it was shown that orally administered L. casei is able to enhance host resistance against oral L. monocytogenes infection. In the gastrointestinal tract, as well as in the spleen and liver, L. monocytogenes numbers were reduced. Furthermore, it is concluded that the enhancement of this anti-Listeria activity might be, at least partly, due to increased cell-mediated immunity. PMID- 11885575 TI - Ultrasonography: is it useful in the diagnosis of cancer in thyroid nodules? AB - The role of ultrasonography (US) in the diagnosis of cancer in thyroid nodules is not well-established. The aim of the present study was to evaluate US performance in predicting cancer in thyroid nodules using a novel approach. Two hundred and eighty-nine patients with thyroid nodular disease were evaluated with clinical, biochemical and cytopathological examinations. Eighty patients with palpable solitary thyroid nodules or multinodular goiters who were to undergo surgery were included, and had a US exam performed by one of us. Some US characteristics of thyroid nodules were associated to cancer: absent halo, hypoechogenicity and microcalcifications, with sensitivity, respectively, of 56, 44 and 56%, and specificity of, respectively, 80, 83 and 94%. These findings were considered positive and were studied in two different combinations: simultaneous, when two or more were positive, and parallel, when any positive finding was present. When positive findings were studied simultaneously, sensitivity ranged 25 to 38% and specificity ranged 89 to 97%. Microcalcifications, associated or not to other findings, were highly specific for thyroid cancer, but they were only present in half of the malignancies. When positive findings were studied in parallel, sensitivity ranged 69 to 81% and specificity ranged 70 to 81%. The parallel combination of hypoechogenicity or microcalcifications or absent halo improved US sensitivity to 81% with an acceptable specificity (70%). This method is potentially useful to help us select patients for surgery when fine-needle aspiration biopsy is repetitively non-diagnostic or select for biopsy incidentally discovered non-palpable nodules. PMID- 11885576 TI - Implications of estradiol and progesterone in pulmonary vasodilatation in cirrhotic patients. AB - The derangement of sex hormone serum levels in cirrhotic patients is well delineated, and increased levels of progesterone and estradiol have been associated to hyperventilation in cirrhotic patients. These hormones have a well known role in the regulation of vascular tone. The aim of this study was to evaluate whether sex hormone levels contribute to pulmonary vasodilatation (PV) and gas exchange abnormalities in cirrhosis. Contrast transesophageal echocardiography, arterial blood gases, parameters of liver function, pulmonary function test, estradiol and progesterone levels were determined in 45 male cirrhotic patients. Nineteen of 45 patients (42.2%) presented PV. Hyperventilation (pressure arterial of CO2< or =35 mmHg) was correlated to progesterone levels (p<0.05) and pressure arterial of CO2 was high in patients with PV (p<0.005) and Child class B and C (p<0.01). Hypoxemia (pressure arterial of O2<80 mmHg) had inverse correlation with progesterone (p<0.05) and estradiol (p<0.05) levels and pressure arterial of O2 was low in patients with Child class B and C (p<0.05). PV was present in patients with high estradiol levels (p<0.05), high progesterone levels (p<0.005) and Pugh class B and C (p<0.05). Logistic regression analysis identified progesterone as the sole independent factor associated to PV (p<0.0005). Multivariate linear regression showed that PV was the sole independent factor related to both pressure arterial of CO2 (p<0.05) and pressure arterial of O2 (p<0.01) levels. PV was independently associated to gas exchange abnormalities in cirrhosis. Progesterone and estradiol were related with PV in cirrhotic patients. PMID- 11885577 TI - Involvement of monoaminergic system in the antidepressant-like effect of the hydroalcoholic extract of Siphocampylus verticillatus. AB - The antidepressant-like effect of the hydroalcoholic extract obtained from aerial parts of Siphocampylus verticillatus, a Brazilian medicinal plant, was investigated in two models of depression in mice and against synaptosomal uptake of serotonin, noradrenaline and dopamine. The immobility times in the forced swimming test (FST) and in the tail suspension test (TST) were significantly reduced by the extract (dose range 100-1000 mg/kg, i.p.), without accompanying changes in ambulation when assessed in an open-field. In addition when given orally the extract was also effective in reducing the immobility time in the TST. The efficacy of extract in the TST was comparable to that of the tricyclic antidepressant imipramine (15 mg/kg, i.p.) and with fluoxetine (32 mg/kg, i.p.). The anti-immobility effect of the extract (600 mg/kg, i.p.) assessed in the TST was not affected by pre-treatment with naloxone (1 mg/kg, i.p., a non-selective opioid receptor antagonist) or L-arginine (750 mg/kg, i.p., a nitric oxide precursor). In contrast, the extract (600 mg/kg, i.p.) antidepressant-like effect was significantly reduced by pre-treatment of animals with p-chlorophenylalanine (PCPA, 100 mg/kg, i.p., an inhibitor of serotonin synthesis), sulpiride (50 mg/kg, i.p., a selective D2 receptor antagonist), prazosin (62.5 microg/kg, i.p., an alpha1 adrenoreceptor antagonist) or by guanosine 5'-monophosphate (GMP, 250 mg/kg, i.p., a nucleotide known to block some actions elicited by NMDA). The biochemical data show that the extract of S. verticillatus inhibited in a graded manner the uptake of monoamines. However, at the IC50 level, the extract was approximately 3.2 to 3.4-fold more potent and also more efficacious in inhibiting the synaptosomal uptake of noradrenaline and serotonin than dopamine. Taken together these data demonstrate that the extract of S. verticillatus elicited a significant antidepressant-like effect, when assessed in the TST and FST in mice. Its action seems to involve an interaction with adrenergic, dopaminergic, glutamatergic and serotonergic systems. PMID- 11885578 TI - Transport rate of arsenic, cadmium, copper and zinc in Potamogeton pectinatus L.: radiotracer experiments with 76As,109,115Cd,64Cu and 65,69mZn. AB - The present study was aimed at obtaining an insight into possible experimental approaches for providing numerical data on both the accumulation of sediment As, Cd, Cu and Zn in the submerged water plant Potamogeton pectinatus L., and the possible corresponding metal flows into the water phase. A hydroculture two compartment system was used as the experimental set-up, and the selected metals were followed by measurements of their radioisotopes 76As, 109Cd, 115Cd, 64Cu, 65Zn and 69mZn. All experiments were performed in single plant mode. The results stress the extreme importance of leakage tests, which were performed using 99mTcO4-, and which resulted in approximately 30% of all experiments being discarded. Metal flows were shown as very near the metal limits of detection or obscured by and/or numerically very near the occurring leakage phenomena. Bio concentration factors BCF (fresh wt. fine root basis) were calculated as 100, 10, 10 and 100-500 l/kg for Cu, Zn, Cd and As, respectively. The mobility, expressed as the shoot/root concentration ratio, CR, was obtained as < 10(-4), < 10(-5), 10(-3) and 10(-3) - 10(-2) for Cu, As, Cd and Zn, respectively. Double-labeling experiments showed that the CR values were due to the exclusive root-mediated transport in radiotracer experiments: results for simultaneous applications of 109Cd and 115Cd or 65Zn and 69mZn showed field-simulated CR values of approximately 0.04 and 14, respectively. Single-tracer experiments, using liquid scintillation counting (LSC) with 109Cd and 65Zn, were shown to strongly improve the sensitivities of flow determinations. Under the applied conditions, metal flows could be determined as <5 x 10(-8), <5 X 10(-8), 3.5+/-1.8 x 10(-10) and <8 x 10(-9) mol/h per kg root fresh wt. for Cu, As, Cd and Zn, respectively. Upscaling calculations, assuming plant steady state behavior, indicate that the metal accumulation in the plants may comprise up to 1% of the sediment metal occurrence, that the major part of an accumulated metal is retained in the plant roots, and that plant-mediated metal flow into the water phase (< 0.01% for Cd, Cu and Zn, < 0.1% for As within a growing season) may be regarded as not significantly contributing to the overall process of metal mobilization. It should be noted, however, that the above conclusions should be drawn with care, due to the pilot nature and the short-term duration of the presented experiments. PMID- 11885579 TI - Persistent organochlorine pollutants in ringed seals and polar bears collected from northern Alaska. AB - Blubber samples from ringed seal (Phoca hispida; n = 8) and polar bear subcutaneous fat (Ursus maritimus; n = 5) were collected near Barrow, Alaska in 1996 as part of the Alaska Marine Mammal Tissue Archival Project (AMMTAP) and retained in the National Biomonitoring Specimen Bank at the National Institute of Standards and Technology in Gaithersburg, Maryland (USA). The samples were analyzed for a variety of persistent organochlorine pollutants (POPs) including polychlorinated biphenyls (PCBs), hexachlorocyclohexanes (HCHs), chlordane and metabolites, hexachlorobenzene (HCB) and DDTs and metabolites. The geometric mean, on a wet mass basis, of sigmaPCBs (sum of 29 congeners and congener groups) were 732+/-282 ng/g (1 S.D.) in seals and 3395+/-1442 ng/g in polar bears. The geometric mean of sigmaDDTs, sigmaHCHs (alpha-, beta- and gamma- HCH) and HCB concentrations (wet mass basis) in seals and bears were 562+/-261 ng/g vs. 74.8+/ 39 ng/g, 380+/-213 ng/g vs. 515 ng/g, and 17.4+/-10.1 ng/g vs. 183+/-153 ng/g, respectively. The geometric mean sum of chlordane (sigmachlordane, sum of cis- and trans-chlordane, cis- and trans-nonachlor, oxychlordane and heptachlor epoxide) and dieldrin concentrations in ringed seals and polar bears were 753+/ 617 ng/g vs. 720+/-315 ng/g and 38.6+/-22.8 ng/g vs. 130+/-65 ng/g, respectively. Apparent bioaccumulation factors (polar bear/ringed seal POP concentrations) were lower in the animals sampled near Barrow, Alaska than in those from locations in the Canadian Arctic. This suggests that polar bears are also preying on marine mammals from lower trophic levels than the ringed seals with correspondingly lower organochlorine levels, such as bowhead whale carcasses. PCB congener patterns in the samples demonstrated the metabolism of certain PCB congeners in the polar bear relative to the ringed seal in agreement with previous studies. Regional comparisons of animals collected in Alaska and Arctic Canada are presented. PMID- 11885580 TI - Acute changes in 3H-PAC and 125I-PYY binding in the nucleus tractus solitarii and hypothalamus after a hypertensive stimulus. AB - Activation of alpha-2-adrenergic and neuropeptide Y (NPY) receptors in the nucleus tractus solitarii (NTS) induces hypotension and bradycardia. On the contrary, activation of angiotensin II (Ang II) receptors leads to hypertension. Acute changes in binding parameters of alpha-2-adrenergic, NPY and Ang II receptors were evaluated in the NTS and paraventricular hypothalamic nucleus (PVN) of rats after a hypertensive stimulus employing quantitative receptor autoradiography. Saturation experiments showed a decrease in the number (Bmax) of alpha-2-adrenergic binding sites in the NTS 6 hours after coarctation-induced hypertension. Furthermore, the affinity of NPY receptors was diminished as seen by the increase in the KD value of 125I-PYY. Tyrosine hydroxylase and NPY immunoreactivities were increased in the NTS and ventral medulla. Binding of 125I Ang II was not changed in the NTS. Binding of all ligands analyzed was not altered in the PVN. The results suggest an acute down-regulation of alpha-2 adrenergic and NPY receptors involved with hypotension in response to hypertensive stimulus, which might be related to an increased availability of catecholamines and NPY in the NTS. PMID- 11885581 TI - From the Centers for Disease Control. Measles--United States, 2000. PMID- 11885582 TI - From the Centers for Disease Control. Revision of guidelines for the prevention of perinatal group B streptococcal disease. PMID- 11885584 TI - From the Centers for Disease Control. Update: influenza activity--United States, 2001-02 season. PMID- 11885583 TI - From the Centers for Disease Control. Status of US Department of Defense preliminary evaluation of the association of anthrax vaccination and congenital anomalies. PMID- 11885585 TI - JAMA patient page. Vehicle safety and children. PMID- 11885586 TI - Social security reform: how might women fare? PMID- 11885587 TI - Effectiveness of smoking cessation initiatives. Smoking cessation services show good return on investment. PMID- 11885588 TI - The declining personal saving rate: is there cause for alarm? PMID- 11885589 TI - Assuring the quality of home care: the challenge of involving the consumer. PMID- 11885590 TI - The deathcare industry. PMID- 11885591 TI - Lifesaving opportunities missed: the challenge of vaccinating older Americans for pneumococcal diseases and influenza. PMID- 11885592 TI - Complementary and alternative medicine: the road less traveled? PMID- 11885593 TI - Retraction: The failure of a once-daily vancomycin dosing regimen in patients with normal renal function. PMID- 11885594 TI - Mitotic exit: closing the gap. AB - Completion of mitosis is triggered by the activation of the Ras-like GTP-binding protein Tem1p. In the November 30, 2001 issue of Cell, Hu et al. suggest that Tem1p activation is achieved by inhibition of its two-component GAP Bub2p/Bfa1p via phosphorylation of Bfa1p by the Polo kinase Cdc5p. Interestingly, activation of spindle checkpoints inhibits Bfa1p phosphorylation, suggesting that these signaling pathways prevent mitotic exit by maintaining the GAP activity of Bub2p/Bfa1p. PMID- 11885595 TI - Plant hormone signaling: getting the message out. AB - Molecular genetic analysis has identified a variety of molecules that are required for correct signaling of the plant hormone abscisic acid (ABA). It now appears that proteins involved in RNA metabolism also modulate the ABA response in Arabidopsis. PMID- 11885596 TI - Thirty-three years later, a glimpse at the ribonuclease III active site. AB - RNase III endonucleases cleave double-stranded RNA, transforming precursor RNAs into mature RNAs that act in pre-mRNA splicing, RNA modification, translation, gene silencing, and the regulation of developmental timing. The recently solved structure of an RNase III endonuclease domain provides a hint at how this family of ribonucleases functions. PMID- 11885597 TI - NHEJ deficiency and disease. AB - In mouse and human, diseases associated with deficiency of DNA ligase IV, a protein involved in DNA double-strand break repair, have been identified. Manifestation of some of these disease phenotypes, namely tumorigenesis, may require additional checkpoint deficiencies. PMID- 11885598 TI - Health and safety issues in an aging workforce. PMID- 11885599 TI - State pharmacy assistance programs 2001: an array of approaches. PMID- 11885602 TI - Consumer-directed services for older people. PMID- 11885603 TI - State taxation of Social Security and pensions in 2000. PMID- 11885604 TI - Beneficial effects of humic acid on micronutrient availability to wheat. AB - Humic acid (HA) is a relatively stable product of organic matter decomposition and thus accumulates in environmental systems. Humic acid might benefit plant growth by chelating unavailable nutrients and buffering pH. We examined the effect of HA on growth and micronutrient uptake in wheat (Triticum aestivum L.) grown hydroponically. Four root-zone treatments were compared: (i) 25 micromoles synthetic chelate N-(4-hydroxyethyl)ethylenediaminetriacetic acid (C10H18N2O7) (HEDTA at 0.25 mM C); (ii) 25 micromoles synthetic chelate with 4 morpholineethanesulfonic acid (C6H13N4S) (MES at 5 mM C) pH buffer; (iii) HA at 1 mM C without synthetic chelate or buffer; and (iv) no synthetic chelate or buffer. Ample inorganic Fe (35 micromoles Fe3+) was supplied in all treatments. There was no statistically significant difference in total biomass or seed yield among treatments, but HA was effective at ameliorating the leaf interveinal chlorosis that occurred during early growth of the nonchelated treatment. Leaf tissue Cu and Zn concentrations were lower in the HEDTA treatment relative to no chelate (NC), indicating HEDTA strongly complexed these nutrients, thus reducing their free ion activities and hence, bioavailability. Humic acid did not complex Zn as strongly and chemical equilibrium modeling supported these results. Titration tests indicated that HA was not an effective pH buffer at 1 mM C, and higher levels resulted in HA-Ca and HA-Mg flocculation in the nutrient solution. PMID- 11885606 TI - Informed consent and other fairy tales. PMID- 11885605 TI - Direct comparison of the impact of head tracking, reverberation, and individualized head-related transfer functions on the spatial perception of a virtual speech source. AB - A study of sound localization performance was conducted using headphone-delivered virtual speech stimuli, rendered via HRTF-based acoustic auralization software and hardware, and blocked-meatus HRTF measurements. The independent variables were chosen to evaluate commonly held assumptions in the literature regarding improved localization: inclusion of head tracking, individualized HRTFs, and early and diffuse reflections. Significant effects were found for azimuth and elevation error, reversal rates, and externalization. PMID- 11885607 TI - English medical law and 'informed consent': an antipodean assessment and alternative. PMID- 11885608 TI - Regulating the reproduction business? PMID- 11885609 TI - Euthanasia: the strength of the middle ground. PMID- 11885610 TI - Physician involvement in a patient's death: a continental European perspective. PMID- 11885611 TI - Decision-making capacity. PMID- 11885612 TI - The role of care assistants in the withdrawal of hydration and nutrition in Germany. PMID- 11885613 TI - The law's treatment of the suicidal. PMID- 11885614 TI - Rights versus risk? Reform of the Mental Health Act 1983. PMID- 11885615 TI - Maintaining a pregnancy following loss of capacity. PMID- 11885616 TI - Easing the passing: end of life decisions and the Medical Treatment (Prevention of Euthanasia) Bill. PMID- 11885618 TI - The last taboo. PMID- 11885619 TI - We have the power. PMID- 11885617 TI - Incompetent patient (adult): Bland and the Human Rights Act 1998. PMID- 11885620 TI - The common good. PMID- 11885621 TI - Gene cheats. PMID- 11885623 TI - Sale of the century: Iceland has come to grief over the human genome. Do we have to? PMID- 11885622 TI - The way ahead. PMID- 11885624 TI - The coming revolution. PMID- 11885625 TI - A right to know? PMID- 11885626 TI - A cure at any cost? PMID- 11885627 TI - Clones are human too. PMID- 11885628 TI - No clones please. PMID- 11885629 TI - Who needs to know? Confusion reigns over the reporting of gene therapy deaths. PMID- 11885630 TI - Without consent. Interview by Charles Seife. PMID- 11885633 TI - Whose rights are they anyway? PMID- 11885634 TI - Fasten your seat belt. PMID- 11885636 TI - They can find you: GPS implants will make it easy to pinpoint people. PMID- 11885637 TI - Rest assured. PMID- 11885638 TI - Will to live? PMID- 11885640 TI - Insurers backtrack on gene tests demands. PMID- 11885639 TI - Don't keep secrets. There's no alternative to being open with the public. PMID- 11885641 TI - Beyond two sexes: boy meets girl. PMID- 11885642 TI - They said it couldn't happen. PMID- 11885643 TI - Beyond two sexes: the gender police. PMID- 11885644 TI - My two mums. Have genetically engineered children arrived by stealth? PMID- 11885645 TI - Born to make you happy. Can stars really be protected against fans intent on cloning them? PMID- 11885646 TI - First person. PMID- 11885647 TI - Lies, damn lies and statistics. Can we trust scientists or are they puppets of the drug industry? PMID- 11885648 TI - Anatomical differences in the peripheral auditory system of mammals and man. A mini review. AB - The major anatomical differences among animal models and man are briefly reviewed. Differences are described in the length and width of the basilar membrane, the number of inner (IHCs) and outer hair cells (OHCs), and the length of cilia on both cell types. Significant differences in the innervation pattern of the IHCs among these species include the number of afferent nerve terminals per IHC, the degree of branching of afferent fibers and the number of synapses per afferent nerve terminal. At the OHCs, the number of afferent and efferent nerve terminals, the presence or absence of presynaptic bodies, reciprocal synapses and the presence of dendrodendritic synapses in the outer spiral bundles may have important physiological functions. In the cochlear nerve, significant differences are described in the number of spiral ganglion cells (SGCs) and cochlear nerve fibers. Furthermore the percentage of myelinated SGCs and the presence of synapses on SGCs varies enormously. PMID- 11885649 TI - Effects of aging on C57BL/6J mice: an electrophysiological and morphological study. AB - Presbycusis is a progressive hearing loss associated with aging that manifests as deafness linked to cochlear morphological degeneration. The effects of aging on the auditory system were studied in C57BL/6J mice using electrophysiological (brainstem auditory evoked potentials; BAEP) and morphological techniques. Cochleae of animals aged 1, 6, 9, 12, 15, 18, 21, or 24 months old were used for that purpose. The BAEP showed a progressive increase in latency and a reduction in amplitude. Morphological studies demonstrated total degeneration of the organ of Corti, which was replaced by a single epithelial layer. An affinity histochemistry study demonstrated minor modifications of glycoconjugates in the organ of Corti during the aging process. PMID- 11885650 TI - Neurotransmission in the human labyrinth. AB - Different neuroactive substances have been found in the efferent pathways of both the olivocochlear and vestibular systems. In the present study, the distribution and role of three neurotransmitters, choline acetyltransferase (ChAT), gamma aminobutyric acid (GABA), and enkephalin were investigated in the human labyrinth of 4 normal-hearing individuals. Immunohistochemical studies in human inner ear research, however, face a problem of procuring well-preserved specimens with maintained neurotransmitter antigenicity and morphology. Methods and findings are reported and discussed. PMID- 11885651 TI - Effects of aging on cochlear monoamine turnover. AB - The aging of the cochlear dopaminergic system has been analyzed by quantifying the levels of dopamine (DA) and its metabolites (3,4-dihydroxyphenylacetic acid, DOPAC, and homovanillic acid, HVA) in adult rats aged 3, 12 or 24 months. The main results were an increase in DA, DOPAC and HVA basal concentrations in aged females with respect to the adults (3 or 12 months old), while just DA and DOPAC increased in aged males. A higher synthesis of DA in aged animals could support these findings, which could indicate some kind of compensatory mechanism related to presbycusis. PMID- 11885652 TI - Degeneration pattern of human first-order cochlear neurons. AB - In the present study, quantitative analysis of the cochlear neurons in the osseous spiral lamina, the modiolus and the internal auditory canal of the same cochlea was performed. Forty-five temporal bones were obtained from 25 patients and prepared by means of microdissection. Ten patients had age-related normal hearing (ARNH) assuming that the 5 children without audiogram had normal hearing. Fifteen patients had sensorineural hearing loss due to various causes. The present study has shown that in young individuals the numbers of cochlear neurons are almost identical at all 3 sites. In patients over 60 years with ARNH, the loss of peripheral nerve processes is always severer than the loss of central nerve processes. This finding suggests that the central processes degenerate at a much slower rate or not at all. Furthermore, 4 different peripheral degeneration patterns were described. The factors responsible for the different degeneration behaviors are still not understood and need further investigation. PMID- 11885653 TI - The inner hair cell afferent/efferent synapses revisited: a basis for new therapeutic strategies. AB - Within the cochlea, the sensory inner hair cells, which transduce the mechanical displacement of the basilar membrane into neural activity, release glutamate that acts on postsynaptic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor channels located on dendrites of primary auditory neurons. Up to now, it has been thought that the auditory nerve responses passively reflected the motion of the basilar membrane supporting the organ of Corti. Here, we show that dopaminergic lateral olivocochlear efferents drive a permanent gain control at the site of auditory action potential initialization. A dysfunction of this system leads to the development of early signs of excitotoxicity. With the knowledge of the molecular mechanisms involved at this first synaptic complex in the cochlea, it is now possible to envisage local treatments for spiral ganglion neurons, either to stop an excitotoxically induced hyperexcitability (probably the starting point of most posttraumatic tinnitus) or to prevent neuronal death (neural presbycusis). PMID- 11885654 TI - Neurotransmission of the cochlear inner hair cell synapse--implications for inner ear therapy. AB - The cochlear inner hair cells (IHCs) are connected to afferent type I auditory neurons and use probably L-glutamate as a neurotransmitter. This IHC synapse receives efferent input from the lateral part of the efferent olivocochlear system with neurons originating in the brainstem and terminating below IHCs synapsing with the afferent type I dendrites. A number of substances have been proposed to function as neurotransmitter or neuromodulator in the lateral efferent system: acetylcholine, gamma-aminobutyric acid (GABA), dopamine, enkephalin and dynorphin. With the aid of microiontophoretic techniques, we studied several transmitter candidates and characterized their receptor subtypes as well as their function on spontaneous or evoked activity of afferent dendrites. The results showed that the glutamatergic transmission of IHCs is facilitated by all types of glutamate receptors: ionotropic glutamate receptors of the N-methyl-D-aspartic acid (NMDA) and a-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) type as well as group I and II metabotropic glutamate receptors. This excitatory glutamatergic transmission is under inhibitory control of GABA (mediated by GABA(A) receptors) and dopamine (mediated by D1 and D2 receptors). In contrast, acetylcholine was able to excite afferent dendrites via muscarinic receptors. These results demonstrate that the lateral efferent system has modulatory function on the glutamatergic neurotransmission of IHCs. Excitation of afferent dendrites by glutamate released from IHCs can thus be tuned in different physiological or pathophysiological conditions. This could have therapeutic implications as it is known that noise exposure is followed by an excitotoxic injury of the IHC synapse. During overexcitation of IHCs, a possible therapy based on the neurochemical data would be (a) glutamate antagonists, (b) dopamine agonists, (c) GABA agonists or a combination from a, b and a, c. PMID- 11885655 TI - Simulation of methods for drug delivery to the cochlear fluids. AB - The inner ear fluids are remarkably 'unstirred' so that it cannot be assumed that applied drugs are dispersed throughout the fluid spaces. Calculation of the effective concentrations achieved when drugs are applied directly to the inner ear is made possible by simulations combining the physical processes involved in solute dispersal, which are diffusion, longitudinal fluid flow and clearances to other compartments. The approach has been validated in numerous experiments in which ion-selective electrodes were used to characterize the spread of marker substances in the cochlear fluids. The model incorporates the known size and geometry of cochlear fluid spaces for 6 species, including the guinea pig and the human. The simulator allows the dispersal of drugs or other substances to be approximated with knowledge of relatively few parameters. The simulation program is available on the Internet at http://oto.wustl.edu/cochlea/. PMID- 11885656 TI - Clinical experience with caroverine in inner ear diseases. AB - The glutamatergic synapses between the cochlear inner hair cells and their afferent neurons seem to be mostly involved in the pathophysiology of the cochlea. Glutamatergic neurotoxicity is characterized by a mitochondrial overproduction of free oxygen radicals damaging lipid membranes and DNA structures of the postsynaptic neuron followed by the clinical symptoms of hearing loss and tinnitus. In preclinical tests, quinoxaline derivatives antagonized these deleterious consequences of too high an amount of free radicals. Therefore the clinically available quinoxaline dione caroverine provides a new approach to a successful treatment of tinnitus, sudden hearing loss and speech discrimination disorders in presbyacusis. The results of corresponding clinical trials are presented. PMID- 11885657 TI - Different action of memantine and caroverine on glutamatergic transmission in the mammalian cochlea. AB - Glutamate is the major transmitter candidate between inner hair cells and the afferent neurons of the mammalian cochlea. We investigated the action of memantine (1-amino-3,5-dimethyl-adamantane) and the quinoxaline derivative caroverine [1-diethylaminoethyl-3,8-(p-methoxybenzyl)-1,2-dihydro-quinoxaline dione] on the glutamatergic transmission in the guinea pig cochlea utilizing extracellular recording techniques and microiontophoretic ejection of substances. While memantine was able to inhibit the NMDA (N-methyl-D-aspartate)-induced firing, the AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) stimulated activity was unaffected. In contrast, caroverine could block both NMDA as well as AMPA-induced firing. As memantine and caroverine are currently in clinical use, these substances could be introduced to the treatment of several cochlear disorders. PMID- 11885658 TI - Role of substance P in the peripheral vestibular and auditory system. AB - The central role of substance P (SP) has attracted growing interest in the past two decades. One of the important physiological functions of SP and other tachykinins is that of a neurotransmitter in primary afferent neurons. Recent immunocytochemical, biochemical and electrophysiological investigations on various neurotransmitters support the hypothesis that SP has a similar function in the vestibular and auditory systems of all mammals including humans. The purpose of this review is to give an overview of the distribution and concomitant physiological functions of this peptide in these sensory systems. PMID- 11885659 TI - Calcium channels in mouse hair cells: function, properties and pharmacology. AB - Adult inner hair cells (IHCs) possess voltage-activated Ca2+ currents that couple receptor potentials to transmitter release at the afferent synapses. Before the onset of hearing both IHCs and outer hair cells (OHCs) exhibit Ca2+ currents. More than 90% of neonatal hair cell (HC) currents flow through alpha1D Ca2+ channel subunits because they are absent in both IHCs and OHCs from alpha1D-/- mice and residual currents are insensitive to L-type agonists. Since lack of the alpha1D-subunit leads to HC degeneration and profound deafness, class D L-type Ca2+ currents seem to be crucial for the development and functioning of the inner ear. Neonatal HC Ca2+ currents were studied using the whole-cell patch clamp technique. They showed rapid activation, rapid deactivation and very little inactivation. They started activating as negative as -65mV. In contrast to alpha1C-mediated (classical L-type) Ca2+ currents, they showed a rather low sensitivity to various L-type antagonists. 10 microM nifedipine e.g. blocked HC Ca2+ currents by about 40% whereas class C L-type Ca2+ currents are completely blocked by 100nM nifedipine. The L-type channel agonist Bay K 8644 increased the HC Ca2+ current by 100-200% and shifted the IV curve to more negative potentials which is similar to its effects in alpha1C-mediated Ca2+ currents. PMID- 11885660 TI - Adrenergic and muscarinic control of cochlear endolymph production. AB - The transduction of sound into nerve impulses in the cochlea is dependent on the stria vascularis. It is a multilayered epithelium, which is part of the epithelial barrier between endolymph and perilymph. The current model designed to explain the generation of the endocochlear potential assumes that the molecular mechanism for the generation of the endocochlear potential is the K(IR)4.1 K+ channel localized in the intermediate cells and that strial marginal cells play an indirect role in the generation of the endocochlear potential. This role is limited to the maintenance of a low K+ concentration in the intrastrial space by absorbing K+ from this space and secreting it into the endolymph. The molecular mechanisms for K+ secretion by strial marginal cells are well established. Strial marginal cells absorb K+ from the intrastrial space via the Na+-K+-ATPase and the Na+2Cl-K+ cotransporter and secrete it across the apical membrane via the IsK/KvLQT1 K+ channel. K+ secretion by strial marginal cells is not only required for the maintenance of the endocochlear potential and to provide the charge carrier for the transduction mechanism, but also to maintain a constant volume of endolymph. Thus, the presence of multiple control mechanisms regulating the rate of K+ secretion is likely. Recent observations suggest that the rate of K+ secretion in strial marginal cells is stimulated by beta1-adrenergic receptors and inhibited by M3 and/or M4 muscarinic receptors. PMID- 11885661 TI - Cochlear blood flow regulation. AB - The regulation of cochlear blood flow is crucial for auditory function due to the sensitivity of this sensory organ to hypoxia. Part of the regulation of cochlear blood flow occurs in the spiral modiolar artery, which provides the main blood supply to the cochlea. Blood flow in general is most effectively regulated through the control of the vascular diameter. The vascular diameter is determined by the degree of constriction of the smooth muscle cells in the vascular wall. A constriction of the smooth muscle cells reduces the diameter of the vascular lumen and thereby decreases blood flow, whereas a relaxation of the smooth muscle cells increases blood flow. The degree of constriction of the smooth muscle cells in the spiral modiolar artery is carefully controlled and must be adjusted properly to the demands of the cochlear tissues. To achieve proper control, smooth muscle cells integrate information from various sources. Vasoconstrictors and dilators may originate from the innervation surrounding the vessel, from endothelial cells lining the vascular lumen or from the smooth muscle cells themselves. Recent advances revealed that smooth muscle cells from different arterioles differ widely in their endowment with mechanisms that regulate the degree of smooth muscle cell tone. Signal transduction mechanisms, which mediate these neurogenic, local and paracrine regulations of smooth muscle contractility are now beginning to be understood. This report reviews recently obtained evidence for adrenergic regulation of cochlear blood flow and then focuses on a novel vasodilation mechanism that involves ryanodine receptors, Ca2+ sparks and the activation of Ca2+-activated K+ channels. PMID- 11885662 TI - ETA receptors in the gerbil spiral modiolar artery. AB - A reduction of blood flow in the spiral modiolar artery (SMA), which supplies the cochlea, is implicated in hearing loss and tinnitus. Endothelins are known to be the most potent endogenous vasoconstrictors. The purpose of the present study was to determine whether the SMA responds to endothelin, which receptor type is present and which signal transduction pathway is involved. The SMA was isolated from the gerbil cochlea by microdissection and superfused in a bath chamber on the stage of an inverted microscope. The vascular diameter was measured by video microscopy, and the cytosolic Ca2+ concentration was monitored simultaneously by fluo-4 fluorescence microscopy. ET-1 and ET-3 caused a dose-dependent vasoconstriction with ET-1 being the more potent agonist. The agonist sarafotoxin S6c had no significant effect. The preferential ET(A) receptor antagonist BQ123 had a higher affinity inhibiting the ET-1-induced vasoconstriction than the preferential ET(B) receptor antagonist BQ788. The ET-1-induced vasoconstriction was prevented by inhibition of phospholipase C with U73122. Blockade of the inositol 1,4,5-trisphosphate (IP3) receptor on the sarcoplasmic reticulum Ca2+ stores with 2-aminoethoxydiphenyl borate and depletion of Ca2+ stores by inhibition of the sarcoplasmic Ca2+-ATPase with thapsigargin prevented ET-1 induced cytosolic Ca2+ increase and reduced the ET-1-induced vasoconstriction. These results demonstrate that endothelin causes a vasoconstriction of the SMA, which is mediated via ET(A) receptors. The data suggest that the signal transduction pathway of the ET(A) receptor involves phospholipase C, IP3 receptors and release of Ca2+ from thapsigargin-sensitive Ca2+ stores. PMID- 11885663 TI - Immunological damage to the inner ear: current and future therapeutic strategies. AB - There is considerable evidence to suggest that hearing and vestibular function can be influenced by immunity in the inner ear. While immunity can protect against infections of the labyrinth, immune response also has the capacity to damage the delicate tissues of the inner ear. Antigenic challenge of the inner ear of sensitized animals leads to rapid accumulation of leukocytes, antibody production, hearing loss and tissue damage. Moreover, a number of systemic autoimmune disorders include hearing loss and vertigo as part of their constellation of symptoms. It also appears that autoimmune damage can exist as an entity confined to the labyrinth. Immune disorders of the inner ear are of special interest since they are among the few forms of hearing loss that are currently amenable to medical treatment. In addition, recent developments in understanding the intracellular pathways that participate in damage to the inner ear provide new opportunities for pharmacotherapy of immune-mediated disorders of hearing and balance. PMID- 11885664 TI - Pharmacological influence on inner ear endothelial cells in relation to the pathogenesis of sensorineural hearing loss. AB - Despite an increasing incidence of acute sensorineural hearing loss, the pathogenesis of this disease remains uncertain. While viral infection of the stria vascularis, organ of Corti or spiral ganglion cells is discussed in the American literature, a vascular genesis with resulting impaired perfusion of the inner ear is favoured by European investigators. Although both hypotheses are supported by different therapeutic strategies to regain normal hearing, the influence of spontaneous remission remains unclear. This study aims at combining these seemingly opposing concepts with the assumption of an immunologically mediated vasculitis with consequent cochlear hypoperfusion. We already know from other organs that during viral vasculitis circulating immunoglobulins are deposited perivascularly, which leads to a local decrease in perfusion and tissue hypoxia. Also in autoimmune diseases, perivasculitis is common with the endothelium playing a major role at the initial stages of the disease. These endothelial cells promote vasculitis by secreting pro-inflammatory cytokines like IL-1, IL-6 or TNF-alpha in addition to the expression of adhesion molecules. Due to the persistence of these immunopathological mechanisms stenosis or atresia with ischaemic necrosis results. To examine whether this pathomechanism is also important in inner ear dysfunction, the immunological response after stimulation of the cochlear endothelium of guinea pigs was determined. In addition, the influence of corticosteroids on this immune cascade was examined. PMID- 11885665 TI - Mpv37 mouse strain--a model for the relationship between the kidney and the inner ear. AB - Using the Mpv17-negative mouse strain, which developed inner ear and kidney dysfunction, we confirm a strong relationship between the kidney and the inner ear. Both organs have specialized epithelia involved in active ion transport, which are separated from the vessels by a basement membrane of similar composition. Our recent results indicate that the glomerular and the stria vascularis basement membrane are simultaneously affected in early stages. Concomitant deposits of IgG during the progressive development of the disease support the idea of a shared antigen. Understanding the pattern of the development of the degeneration will provide a basis towards understanding the essential role of the Mpv17 protein in the structures of both organs and may provide a basis for future therapeutic intervention. PMID- 11885666 TI - Sensitivity to glutamate neurotoxicity in different developmental periods of the rat cochlea. AB - Cochlear neurotoxicity induced by the intraperitoneal administration of monosodium glutamate (MSG) has been analyzed during the postnatal development of the auditory receptor of the rat. The animals were treated with MSG during two postnatal periods. The electrophysiological recordings showed that MSG treatment produced a decrease in the 8th nerve compound action potential. The effect was more marked in the animals treated between the 9th and 12th postnatal day than in the others, with a qualitative decrease in neuronal density in the spiral ganglion. These results suggest that there is a period of maximum sensitivity to the cochlear neurotoxicity induced by MSG in the postnatal development of the rat. PMID- 11885667 TI - Protective mechanisms of sound conditioning. AB - Evidence continues to accumulate demonstrating the importance of reducing the deleterious effects of noise trauma by sound conditioning. Sound conditioning is an active process induced by low-level, nondamaging noise exposure that creates long-term protective effects to subsequent detrimental forms of noise trauma. This phenomenon is now shown to occur in a variety of mammals, including gerbils, chinchillas, guinea pigs, rabbits, rats, mice and human subjects. Different sound conditioning paradigms have been proven successful in preventing pathological changes to the auditory system. These studies are reviewed in the present chapter and the possible biological mechanisms underlying this phenomenon are discussed. PMID- 11885668 TI - Dentistry for people with special needs--a perspective from Australia and New Zealand. PMID- 11885670 TI - Selection of restorative materials for the atraumatic restorative treatment (ART) approach: a review. AB - The atraumatic restorative treatment (ART) technique or approach for the restoration of primary and permanent teeth has been widely adopted in, but not limited to, developing countries. However, the requirement for the placement of the restorative materials under often less-than-ideal conditions imposes significant restrictions on their selection; and there have been very few randomized clinical trials or reports comparing different types of restorative materials and treatments. Although conventional glass-ionomer cements (GICs) have relatively poor mechanical and adhesive strengths, their satisfactory biological features, ease of use, and low costs are distinct advantages. Most of the published reports of the clinical performance of the newer, high-strength esthetic conventional GICs specifically marketed for the ART approach have been from short-term studies. Satisfactory clinical performance has been demonstrated for single-surface posterior restorations only, over three years. Findings indicate that further improvements in restorative materials are still required for their use with the ART approach, together with further clinical investigations of the remineralization of shallow open caries lesions, as an alternative to placing definitive restorations. PMID- 11885669 TI - Coping with tooth pain: a qualitative study of lay management strategies and professional consultation. AB - This research analyzes transcripts of semi-structured interviews with patients presenting with tooth pain at a rural dental clinic in North Florida. The primary objectives are to identify the strategies patients use to manage their pain and to elucidate the decision-making process leading to the clinic visit. Although respondents understood that their condition was not self-limiting, only about one half contacted the clinic within several days of the onset of their pain. Most tried one or more lay management strategies. PMID- 11885671 TI - The implications of visual impairment in an elderly population in recognizing oral disease and maintaining oral health. AB - The incidence of impairment and disability increases in the elderly population. Disability can affect elderly people's ability to maintain oral health, maintain access to dental care, and accept dental treatment. The oral health of visually impaired people can be disadvantaged, since they are not in a position to detect and recognize early oral disease and may be unable to take immediate action unless informed of the situation. The aim of this pilot study was to identify the problems experienced by a visually impaired elderly population regarding the maintenance of oral health and the need to seek treatment. Sixty-two legally blind people underwent a structured interview and a clinical dental examination. Data are presented descriptively. All the participants would be potentially able to maintain their own adequate oral health level if given the appropriate stimulus. At the time of the study, 53% brushed their own teeth, 39% of whom brushed daily. Of the denture wearers, 58% cleaned them at least once a week. Most of the participants were independent. Eighty-two percent believed that they did not need help to brush their teeth or dentures, even though 85% had never been shown how to brush their teeth. Eighty percent of people did not realize that regular oral reviews were necessary. Other barriers to regular care included poor health, mobility problems, cost, and fear. Twenty-one percent of the sample had toothache or a denture problem. The professionally assessed treatment need was high in the dentate group, and 32% of denture wearers had denture-related pathology. The professionally assessed treatment need was greater than the visually impaired people's perceived need for care. The majority of visually impaired elderly were capable of maintaining a reasonable level of oral hygiene by themselves and were aware of their own dental needs but either had no reason to seek care or were unable to access oral health care services. It is important for visually impaired persons to realize that they need supervision to maintain good oral hygiene standards and to ensure the early identification of oral pathology. Also, the barriers to access to oral health services need to be reduced. PMID- 11885673 TI - A novel multi-degree-of-freedom thick-film ultrasonic motor. AB - This paper describes a new multi-degree-of-freedom (MDOF) ultrasonic motor that comprises few parts and is based on low-cost thick-film technology. Conventional ultrasonic motors using bulk lead zirconate titanate (PZT) or thin-film PZT layers are relatively expensive at the present time. Thick-film printed PZT technology provides the opportunity to reduce the costs of ultrasonic motors. To demonstrate the feasibility of this approach, an ultrasonic motor was fabricated from alumina using thick-film printed PZT actuators. The thick-film PZT and electrode layers were printed on a thin alumina plate, and a tiny cylinder was mounted at its center. This cylinder magnifies the lateral displacement of the stator, holds the spherical rotor, and transmits the driving force to the sphere. Three bending vibrations, B22, B30, B03, of the plate were applied to rotate the sphere. Sufficient displacements for rotating the sphere were obtained near the resonance of B22 by applying an excitation voltage of 200 V peak-to-peak via a three-phase drive circuit. Rotations in three orthogonal directions have been observed by controlling the phase of the driving signal to the PZT electrodes, and a MDOF ultrasonic motor was successfully realized. PMID- 11885672 TI - Oral health profile in an institutionalized population of Italian adults with mental retardation. AB - The oral health of 219 residents with mental retardation living in a long-term care institution near Milan was assessed. The dental and periodontal status, daily habits, oral hygiene, and oral mucosal status were evaluated. Of the sample, 179 (81.7%) were males. The mean age of the residents was 61.3 years, and the degree of cooperation was evaluated as good for 131 subjects (59.8%), fair for 79 (36.1%), and poor for nine (4.1%). The percentage of residents who were edentulous was 21.5% (47 subjects), of whom 28 subjects (59.6%) were without dentures. Evaluation showed an overall DMFT of 23.1, and the average number of missing teeth was 20.5. All subjects had periodontal disease: Forty-five subjects had calculus and/or shallow pockets (4-5 mm); 61 had deep pockets (> or = 6 mm). The most common mucosal lesion was oral stomatitis (49.3%). These findings underline the need for special programs aimed at institutionalized subjects with mental retardation. PMID- 11885674 TI - An accurate model for capacitive micromachined ultrasonic transducers. AB - Modeling of capacitive micromachined ultrasonic transducers (cMUTs) is based on a two-port network with an electrical and a mechanical side. To obtain a distributed model, a solution of the differential equation of motion of the diaphragm for each element of the transducer has to be found. Previous works omit the mechanical load of the cavity behind the diaphragm, i.e., the effect of the gas inside. In this paper, we propose a distributed model for cMUTs that takes this effect into account. A closed-form solution of the mechanical impedance of the membranes has been obtained, including the effect of the restoring forces because of the stiffness of the membrane and because of the compression of the air in the cavity. Simulation results based on the presented model are compared with the experimental data for two types of cMUTs reported in the recent literature. It is demonstrated that the compression of the air has a significant effect on the fundamental frequency of the air transducer, with a deviation of about 22% from the prediction of a model that does not consider the interaction between the vibrating diaphragm and the air cushion. PMID- 11885675 TI - Numerical simulation of the electro-acoustical response of a transducer excited by a time-varying electrical circuit. AB - Existing methods for the modeling of piezoelectric transducer response are generally frequency domain-based. The major disadvantage of this type of model is that they cannot take into account the electrical elements present in the emitting or receiving circuit whose values vary with respect to time. The need for a method that accounts for time-varying elements arises, for example, when the circuit comprises active electrical elements, such as diodes, or when the transducer is excited by capacitive discharge via a switch. Indeed, in this last example, it is known that the output impedance of the generator depends on the state of the switch: if it is off, its value is high; if it is on, its value is low. A time domain-based method is presented to compute the electro-acoustical response of a piezoelectric transducer and its electrical circuit, taking into account the presence of time-varying elements. An application to a current example makes it possible to show the influence of these elements on waveforms and the capacity of our model to account for them. PMID- 11885676 TI - Present and future of piezoelectric single crystals and the importance of B-site cations for high piezoelectric response. AB - High quality piezoelectric single crystals, such as Pb(Zn(1/3)Nb(2/3))O3-PbTiO3 (PZNT) and Pb(Mg(1/3)Nb(2/3))O3-PbTiO3 (PMNT), have been investigated, and, because their piezoelectric properties are greatly superior to those of Pb(Zr(1 x)Ti(x))O3 (PZT) ceramics, they have been used for certain transducer applications since the late 1990s. The present situation for these relaxor-PT (lead titanate) single crystals is summarized. In this review, some possible high Tc > 200 degrees C single crystals are also introduced. Single crystals of Pb(In(1/2)Nb(1/2))O3-PbTiO3 (PINT) binary system and Pb(Mg(1/3)Nb(2/3))O3 Pb(Sc(1/2)Nb(1/2))O3-PbTiO3 (PSMNT) tertiary system have been synthesized, and their electrical properties are reported. In addition, a novel guiding principle for discovering excellent piezoelectric materials, namely the presence of low molecular mass B-site ions that can enter the lead-perovskite Pb(B'B'')O3 structure, is introduced. PMID- 11885677 TI - Optical measurement of ultrasonic Poynting and velocity vector fields. AB - This report describes a method for estimating several wide bandwidth ultrasonic field parameters from optical measurements of the local, acoustically induced, refractive index perturbation in water. These parameters include Poynting and particle velocity vector fields as well as pressure and density fields at any temporal delay under mild (forward-propagating) assumptions on the angular plane wave spectrum of the ultrasound field. A sampling theorem is derived stating that two complete measurements of the three-dimensional pressure field separated in time by delta t allow release of the forward-propagating assumption for every acoustic wave number k satisfying k not = n pi/(c delta t), where c is the acoustic wave speed in the medium and n an integer greater than zero. The approach provides detailed measurements of very general ultrasound fields. Two optical measurement methods that acquire the Radon transform of the three dimensional refractive index perturbation are briefly reviewed. It is shown that the Radon transform of the field itself satisfies a two-dimensional wave equation and may be propagated independently forward or backward in time under a source free model. Conversely, the Radon transform of the ultrasound field measurement at several known time delays provides a means of applying a filter to the data based on known ultrasound propagation models. Each two-dimensional distribution may be propagated to a common time point and the ensemble averaged, thus incorporating the propagation model into the measurement. We support the presented theory with several experiments. PMID- 11885678 TI - Clutter filter design for ultrasound color flow imaging. AB - For ultrasound color flow images with high quality, it is important to suppress the clutter signals originating from stationary and slowly moving tissue sufficiently. Without sufficient clutter rejection, low velocity blood flow cannot be measured, and estimates of higher velocities will have a large bias. The small number of samples available (8 to 16) makes clutter filtering in color flow imaging a challenging problem. In this paper, we review and analyze three classes of filters: finite impulse response (FIR), infinite impulse response (IIR), and regression filters. The quality of the filters was assessed based on the frequency response, as well as on the bias and variance of a mean blood velocity estimator using an autocorrelation technique. For FIR filters, the frequency response was improved by allowing a non-linear phase response. By estimating the mean blood flow velocity from two vectors filtered in the forward and backward direction, respectively, the standard deviation was significantly lower with a minimum phase filter than with a linear phase filter. For IIR filters applied to short signals, the transient part of the output signal is important. We analyzed zero, step, and projection initialization, and found that projection initialization gave the best filters. For regression filters, polynomial basis functions provide effective clutter suppression. The best filters from each of the three classes gave comparable bias and variance of the mean blood velocity estimates. However, polynomial regression filters and projection-initialized IIR filters had a slightly better frequency response than could be obtained with FIR filters. PMID- 11885679 TI - A 30-MHz piezo-composite ultrasound array for medical imaging applications. AB - Ultrasound imaging at frequencies above 20 MHz is capable of achieving improved resolution in clinical applications requiring limited penetration depth. High frequency arrays that allow real-time imaging are desired for these applications but are not yet currently available. In this work, a method for fabricating fine scale 2-2 composites suitable for 30-MHz linear array transducers was successfully demonstrated. High thickness coupling, low mechanical loss, and moderate electrical loss were achieved. This piezo-composite was incorporated into a 30-MHz array that included acoustic matching, an elevation focusing lens, electrical matching, and an air-filled kerf between elements. Bandwidths near 60%, 15-dB insertion loss, and crosstalk less than -30 dB were measured. Images of both a phantom and an ex vivo human eye were acquired using a synthetic aperture reconstruction method, resulting in measured lateral and axial resolutions of approximately 100 microm. PMID- 11885680 TI - 1.5-D high intensity focused ultrasound array for non-invasive prostate cancer surgery. AB - The aim of this study is to demonstrate the feasibility of a new spherically curved 1.5-D phased array for the treatment of localized prostatic cancer. The device is designed to conform to the Ablatherm machine (EDAP-Technomed, France), a commercially available machine in which high intensity focused ultrasound (HIFU) treatment for prostate cancer is administered transrectally. It uses high intensity electronically focused ultrasound to steer a beam along two axes, allowing enough depth to be reached to treat large prostates and eliminating two degrees of mechanical movement. Through computer simulation, it was determined that a curved 1.5-D configuration offered the optimal design. Two configurations were then proposed, and their ability to steer a beam within a target volume centered on the geometric focus of the transducer was simulated. An eight-element prototype was constructed to test the piezo-composite material and its electro acoustical efficiency. Then, an array was constructed, and a multi-channel amplifier and control system were added, to permit remote operation. Acoustical and electrical measurements were made to verify performance. Finally, the 1.5-D array was tested in vitro on samples of pig liver to confirm the ability to induce lesions. PMID- 11885681 TI - A sigma-delta-based sparse synthetic aperture beamformer for real-time 3-D ultrasound. AB - Sigma-delta modulation allows delay resolution in ultrasound beamformers to be achieved by simple clock cycle delays applied to the undecimated bit-stream, greatly reducing the complexity of the signal processing and the number of bits in the datapath. The simplifications offered by this technique have the potential for low power and portable operation in advanced systems such as 3-D and color Doppler imagers. In this paper, an architecture for a portable, real-time, 3-D sparse synthetic aperture ultrasound beamformer based on sigma-delta modulation is presented, and its simulated performance is analyzed. Specifically, with a 65 element linear phased array and three transmit events, this architecture is shown to achieve a 1.1 degrees beamwidth, a -54-dB secondary lobe level, and a theoretical frame rate of 1700 frames/s at lambda/64 delay resolution using a second-order low pass sigma-delta modulator. Finally, a technique for modifying the proposed multi-beam architecture to allow improved analog-to-digital (A/D) resolution by premodulating the input signal for bandpass sigma-delta modulation is also presented. PMID- 11885682 TI - Experimental and simulated ultrasonic characterization of complex damage in fused silica. AB - The growth of a laser-induced, surface damage site in a fused silica window was monitored by the ultrasonic pulse-echo technique. The laser damage was grown using 12-ns pulses of 1.053-microm wavelength light at a fluence of approximately 27 J/cm2. The ultrasonic data were acquired after each pulse of the laser beam for 19 pulses. In addition, optical images of the surface and subsurface damage shape were recorded after each pulse of the laser. The ultrasonic signal amplitude exhibited variations with the damage size, which were attributed to the subsurface morphology of the damage site. A mechanism for the observed ultrasonic data based on the interaction of the ultrasound with cracks radiating from the damage site was tested using two-dimensional numerical simulations. The simulated results exhibit qualitatively similar characteristics to the experimental data and demonstrate the usefulness of numerical simulation as an aid for ultrasonic signal interpretation. The observed sensitivity to subsurface morphology makes the ultrasonic methodology a promising tool for monitoring laser damage in large aperture laser optics used in fusion energy research. PMID- 11885683 TI - Design optimization of beam-like ferroelectrics-silicon microphone and microspeaker. AB - Design optimization of beam-like ferroelectrics-silicon integrated microphone and microspeaker is studied. The Pt/Pb(Zr,Ti)O3/Pt/SiO2/Si3N4 cantilever structure is designed theoretically using multimorph model. The sensitivity and the sound pressure level (SPL) of the integrated microphone and microspeaker are calculated. It is found that the sensitivity of the microphone and the SPL of the microspeaker are very high. This integrated microphone and microspeaker will be valuable for high quality micro-acoustic devices. PMID- 11885684 TI - Lumen pressure within obliquely insonated absorbent solid cylindrical shells with application to Doppler flow phantoms. AB - Flow phantoms used in medical ultrasound usually employ a plastic tube as a blood vessel mimic. These tubes often have acoustic properties differing significantly from the tissue and blood-mimicking media, which results in distortion of the acoustic pressure field within the tubes and, hence, of the Doppler flow spectra. Previous analyses of this problem have used some form of the infinite plate transmission coefficient, although at least one ray-based analysis has considered a cylindrical interface but with zero wall thickness. In this paper, we compare these approximate pressure fields with the exact solution for oblique incidence on a viscoelastic cylindrical shell at 5 MHz to find for which materials the plate approximation is valid. The shell has water both inside and outside, but it can be modified to use a different fluid inside and also to include absorption in either fluid. We find the plate approximation is reasonable for soft tubes such as the copolymer Cflex (Cole-Palmer, Niles, IL) but much less so for hard tubes such as polymethylmethacrylate (PMMA). PMID- 11885685 TI - Two-dimensional ultrasonic strain rate measurement of the human heart in vivo. AB - A study is presented in which the feasibility of two-dimensional strain rate estimation of the human heart in vivo has been demonstrated. To do this, ultrasonic B-mode data were captured at a high temporal resolution of 3.8 ms and processed off-line. The motion of the RF signal patterns within the two dimensional sector image was tracked and used as the basis for strain rate estimation. Both axial and lateral motion and strain rate estimates showed a good agreement with the results obtained by more established, one-dimensional techniques. PMID- 11885687 TI - Evidence for a common molecular basis for sequence recognition of N3-guanine and N3-adenine DNA adducts involving the covalent bonding reaction of (+)-CC-1065. AB - The antitumor antibiotic (+)-CC-1065 can alkylate N3 of guanine in certain sequences. A previous high-field 1H NMR study on the (+)-CC-1065d[GCGCAATTG*CGC]2 adduct (* indicates the drug alkylation site) showed that drug modification on N3 of guanine results in protonation of the cross-strand cytosine [Park, H.-J.; Hurley, L. H. J. Am. Chem. Soc. 1997, 119, 629.]. In this contribution we describe a further analysis of the NMR data sets together with restrained molecular dynamics. This study provides not only a solution structure of the (+) CC-1065(N3-guanine) DNA duplex adduct but also new insight into the molecular basis for the sequence-specific interaction between (+)-CC-1065 and N3-guanine in the DNA duplex. On the basis of NOESY data, we propose that the narrow minor groove at the 7T8T step and conformational kinks at the junctions of 16C17A and 18A19T are both related to DNA bending in the drugDNA adduct. Analysis of the one dimensional 1H NMR (in H2O) data and rMD trajectories strongly suggests that hydrogen bonding linkages between the 8-OH group of the (+)-CC-1065 A-subunit and the 9G10C phosphate via a water molecule are present. All the phenomena observed here in the (+)-CC-1065(N3-guanine) adduct at 5'-AATTG* are reminiscent of those obtained from the studies on the (+)-CC-1065(N3-adenine) adduct at 5'-AGTTA*, suggesting that (+)-CC-1065 takes advantage of the conformational flexibility of the 5'-TPu step to entrap the bent structure required for the covalent bonding reaction. This study reveals a common molecular basis for (+)-CC-1065 alkylation at both 5'-TTG* and 5'-TTA*, which involves a trapping out of sequence-dependent DNA conformational flexibility as well as sequence-dependent general acid and general base catalysis by duplex DNA. PMID- 11885686 TI - Specific cell-signal targets for cancer chemotherapy. AB - Attempts to develop drugs, specific for cancer cells, are dealt here according to the intended cell-target. While many target specific drugs were developed, they reach only moderate successes in clinics for reasons, such as, delivery problem, lack of in vivo efficacy or toxicity. However, recent efforts focusing on the diversity of tyrosine kinases, participating in cell-signal transduction, brought fruit. The firs such drug, Givec, approved by the USFDA recently, is used in clinics with great success to threat CML. The drug inhibits tyrosin kinase of bcr abl, c-abl and v-abl. Work is progressing on other tyrosin kinase inhibitors and on other type of specific cancer cell signal protein inhibitors. These efforts are hoped to yield better cures for cancer in the near future. PMID- 11885688 TI - Synthesis, antimicrobial and molluscicidal activities of new benzimidazole derivatives. AB - A series of benzimidazole Schiff's bases, thiosemicarbazides were synthesized, azole ring systems as 1,3,4-triazole, 1,3,4-oxadiazole were prepared. 1 Methylbenzimidazole incorporated to substituted dithio-carbamate, thiophenol, diethylamine via acetamido group were synthesized. A series of pyrimidinobenzimidazoles, triazinobenz-imidazoles, and 2-(acetonylamino)-1 methylbenzimidazole were prepared. The antimicrobial and molluscicidal activities of some newly prepared compounds were carried out. PMID- 11885689 TI - Synthesis, characterization and in vtro identification of N7-guanine adduct of 2 bromopropane. AB - Recently, we have reported that 2-bromopropane might have an immunotoxic potential in rats when exposed for 28 days. In the present studies, the possibility of 2i-deoxyguanosine adduct formation by 2-bromopropane was investigated in vitro to elucidate molecular mechanism of 2-bromopropane-induced immunosuppression. N7-Guanine adduct of 2'-bromopropane (i.e., N7-isopropyl guanine) was chemically synthesized and structurally characterized by analysis of UV, 1H-NMR, '3C-NMR, COSY and fast atom bombardment mass spectrometry to use as a reference material. Incubation of 2'-deoxyguanosine with an excess amount of 2 bromopropane in PBS buffer solution, pH 7.4, at 37 degrees C for 16 h, followed by a thermal hydrolysis, produced a detectable amount of N7-isopropyl guanine by an HPLC and UV analysis. The present results suggest that 2-bromopropane might form a DNA adduct in N7 position of 2'-deoxyguanosine at a physiological condition. PMID- 11885691 TI - Syntheses of (+/-)-homoepibatidine analogues. AB - Syntheses of (+/-)-homoepibatidine analogues (2), which contain the 8-azabicyclo [3.2.1]octane ring system, were achieved by using palladium-catalyzed reductive coupling reaction from 3 and the analgesic activity was tested by Mouse writhing antinociceptive assay. PMID- 11885690 TI - Stereoselective syntheses of (+/-)-epibatidine analogues. AB - Stereoselective syntheses of (+/-)-epibatidine analogues 2, which contain the 8 azabicyclo [3.2.1]octane ring system, were achieved by using palladium-catalyzed cross-coupling reaction from 4 and the analgesic activity was tested by Mouse writhing antinociceptive assay. PMID- 11885692 TI - A new coumarin from the stem of Angelica dahurica. AB - One new and three known coumarins were isolated from the CHCl3 soluble fraction of Angelica dahurica stem. On the basis of spectral data, the structures of the isolated compounds were determined to be scopoletin, angelol I, angelol H and 6 [(1S), 2(R)-2, 3-dihydroxy-1-methoxy-3-methylbutyl]-7-methoxycoumarin; the latter being isolated for the first time from a plant source. PMID- 11885693 TI - Structure determination of a new lupane-type triterpene, tiarellic acid, isolated from Tiarella polyphylla. AB - A new 27-carboxylic lupane-type triterpene, tiarellic acid (1), was isolated from Tiarella polyphylla together with corosolic acid (2) and tormentic acid (3). Tiarellic acid was characterized as 3,23-dihydroxy-20(29)-lupen-27-oic acid and its NMR data were unambiguously assigned using 2-D NMR techniques. PMID- 11885694 TI - Effect of butanol fraction of Panax ginseng head on gastric lesion and ulcer. AB - From our previous result that Panax ginseng head extract had inhibition of gastric damages, the extract was fractionated. Among the hexane, chloroform, butanol and water fractions, butanol fraction showed the most potent inhibition of HCl.ethanol-induced gastric lesion, aspirin-induced gastric ulcer, acetic acid induced ulcer and Shay ulcer. Butanol fraction showed significant increase in mucin secretion, and inhibited malondialdehyde (MDA) and H+/ K+ATPase activity in the stomach. This results indicate that the effectiveness of the fraction on gastric damages might be related to inhibition of acid secretion, increment of mucin secretion and antioxidant property. PMID- 11885695 TI - Effects of Opuntia ficus-indica var. Saboten stem on gastric damages in rats. AB - The effects of the dried stem powder of Opuntia ficus-indica var. saboten (OF-s) were investigated on gastric lesion and ulcer models in rats. It showed significant inhibition in HCl ethanol-induced gastric lesion at the doses of 200 and 600 mg/kg p.o. and in HCl.aspirin-induced gastric lesion at 600 mg/kg p.o. OF s also showed significant inhibition in indomethacin-induced gastric lesion at the doses of 200 and 600 mg/kg, p.o. However, it did not affect both the aspirin induced and Shay ulcers in rats. It also did not affect gastric juice secretion, acid output and pH. These data indicate that OF-s only possesses pronounced inhibitory action on gastric lesion without antiulcer activity in rats. PMID- 11885696 TI - Epidermis proliferative effect of the Panax ginseng ginsenoside Rb2. AB - Ginseng has been used as a traditional medicine with various therapeutic effects. However, it is still unknown which component of this plant is effective at promoting wound healing. Recently, ginsenoside Rb2 has been reported to improve wound healing. In this study, to investigate the reported wound healing effect of the ginsenoside Rb2, cell morphology and protein factors involved in epidermal formation were evaluated by immunochemical and immunoblotting analysis. Rb2 stimulated epidermal cell proliferation, and the cell showed a 1.5-fold increase in thymidine uptake compared to the control (p<0.05, n=3). Furtheremore, Rb2 was found to stimulate epidermis formation in a dose-dependent manner in raft culture, and to dose dependently enhance the expressions of protein factors related to cell proliferation, namely, epidermal growth factor and its receptor, fibronectin and its receptor, keratin 5/14, and collagenase I (p<0.05, n=3-9). It is believed that ginsenoside Rb2 enhances epidermal cell proliferation by upregulating the expressions of these proliferation-related factors. PMID- 11885697 TI - A radical scavenging farnesylhydroquinone from a marine-derived fungus Penicillium sp. AB - Farnesylhydroquinone (1) has been isolated from the mycelium of a marine-derived fungus of the genus Penicillium. The structure of the compound (1) has been elucidated by spectral method. The compound 1 exhibits potent radical scavenging activity (IC50 12.5 microM) against the DPPH. PMID- 11885698 TI - Spontaneous release of glycosylphosphatidylinositol (GPI)-anchored renal dipeptidase from porcine renal proximal tubules. AB - The incubation of porcine renal proximal tubules (PTs) resulted in the release of the glycosylphosphatidylinositol (GPI)-anchored renal dipeptidase (RDPase, EC 3. 4. 13. 19) from the membrane after a lag period of approximately 6 hours. This spontaneous release of RDPase from the membrane was inhibited by antibiotics. When the incubation supernatant was added back to fresh PTs, both the antibiotic inhibition of RDPase release and the lag period disappeared. The released RDPase reacted with an anti-cross reacting determinant antibody indicating the presence of the Ins (1,2-cyc)P moiety. These results suggest that bacteria in the PTs, when incubated, grow and secrete a phosphatidylinositol-specific phospholipase C (PI-PLC). This enzyme then hydrolyses the GPI-anchored RDPase and is transferable. RDPase was purified following its release from the membrane by this simple and inexpensive method which may also be applied to other GPI-anchored proteins. PMID- 11885699 TI - Enhancement of paracellular transport of heparin disaccharide across Caco-2 cell monolayers. AB - The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested. The cytotoxicity of these enhancers was also examined. The enhancing effects by Quillaja saponin, dipotassium glycyrrhizinate, 18beta-glycyrrhetinic acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, 18beta-glycyrrhetinic acid and taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds. It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium. PMID- 11885701 TI - Now we're talking...but who are we talking about? PMID- 11885700 TI - Korean mistletoe lectin-induced apoptosis in hepatocarcinoma cells is associated with inhibition of telomerase via mitochondrial controlled pathway independent of p53. AB - The extract of European mistletoe (Viscum album, L) has been used in adjuvant chemotherapy of cancer and mistletoe lectins are considered to be major active components. The present work was performed to investigate the effects of Korean mistletoe lectin (Viscum album L. coloratum agglutinin, VCA) on proliferation and apoptosis of human hepatoma cells as well as the underlying mechamisms for these effects. We showed that VCA induced apoptosis in both SK-Hep-1 (p53-positive) and Hep 3B (p53-negative) cells through p53- and p21-independent pathways. VCA induced apoptosis by down-regulation of Bcl-2 and by up-regulation of Bax functioning upstream of caspase-3 in both cell lines. In addition, we observed down-regulation of telomerase activity in both VCA-treated cells. Our results provide direct evidence of the anti-tumor potential of this biological response which comes from inhibition of telomerase and consequent inducing apoptosis. VCA induced apoptosis is regulated by mitochondrial controlled pathway independently of p53. These findings are important for the therapy with preparation of mistletoe because they show that telomerase-dependent mechanism can be targeted by VCA in human hepatocarcinoma. Taken together, our results suggest that the VCA, considered as a telomerase-inhibitor, can be envisaged as a candidate for enhancing sensitivity of conventional anticancer drugs. PMID- 11885702 TI - Low-level lead exposure and children. AB - The adverse effects of environmental lead exposure on the mental development of young children are well established. There is no safe level of blood lead below which children are not affected. Recent research expands our understanding of the impact of lead exposure continuing into later childhood, as well as its effects on children's behaviour. However, social and other environmental factors also contribute to variance in measures of developmental and behavioural outcomes. Lead is associated with only modest effects on children's development, but is a potentially modifiable risk factor. As environmental exposure to lead declines for the whole population, continued specific attention is needed for children living in industrial areas. PMID- 11885703 TI - Slow transit constipation in children. AB - Patients with chronic constipation that fails to respond to treatment remain a challenge for paediatricians and surgeons. Ongoing work in our institution suggests that a number of children with intractable symptoms have slow transit constipation, which has only been described recently in paediatrics. Common features of slow transit are: delayed passage of the first meconium stool beyond 24 h of age, symptoms of severe constipation within a year, or treatment resistant 'encopresis' at 2-3 years, soft stools despite infrequent bowel actions, and delay in colonic transit on a transit study. A proportion of children with slow transit constipation have an abnormality of intestinal innervation associated with the dysfunctional colonic motility, recognized as intestinal neuronal dysplasia (IND). Intestinal neuronal dysplasia type B, the most common variant of IND, is defined on rectal biopsy by hyperplasia of the submucosal plexus. On laparoscopic colon muscle biopsy, many specimens show reduced numbers of excitatory substance P-immunoreactive nerve fibres in the circular muscle. Functional markers of the nerves allow new diagnostic criteria to be developed which may also allow a more rational approach to treatment. The aetiology remains obscure and the optimal management poorly defined, although subtotal colectomy, proximal colostomy or appendicostomy (for antegrade enemas) have been tried. Once the anatomy and physiology of the colon in children with slow colonic transit is better understood, we will have defined not only a new form of constipation, but also will be able to consider new therapies. PMID- 11885704 TI - Childhood speech disorders: reported prevalence, comorbidity and socioeconomic profile. AB - OBJECTIVE: To describe the reported prevalence, comorbidity and socioeconomic status (SES) of children with speech disorders. METHODOLOGY: Data from the 1995 Australian Health Survey were used. Information relating to the health of 12 388 children aged 0-14 years was collected via face-to-face interviews with a responsible adult. Speech disorders were recorded if children had reported difficulty talking, producing speech sounds, or who stuttered. RESULTS: The prevalence of childhood speech disorders (CSD) was 1.7% (n = 209). Of this group, 25.8% (n = 54) had a developmental delay or intellectual impairment; when these were excluded, the prevalence of CSD was reduced to 1.3% (n = 155). Among males, the peak prevalence occurred at age 5 (6.5%), for females the highest rates were for 3-4-year-olds (1.8%). Children with a speech disorder had a greater number of additional health problems. No relationship was found between SES and CSD. CONCLUSIONS: Children with speech disorders often have complex health and developmental needs. Developing effective prevention programs with a view to improving the long-term health and social outcomes of these children will require a mix of individual (clinical) and population-based (public-health) strategies. PMID- 11885705 TI - Abolishing the bag: a quality assurance project on urine collection. AB - OBJECTIVE: To audit practice in diagnosis of urinary tract infection (UTI) and implement a protocol to reduce the number of falsely diagnosed or misse PMID- 11885707 TI - Childhood poisoning in Queensland: an analysis of presentation and admission rates. AB - OBJECTIVE: To determine the presentation rates for paediatric poisoning by ingestion and the determinants of hospital admission. METHODOLOGY: Cross sectional survey using an injury surveillance database from emergency departments in South Brisbane, Mackay and Mt Isa, Queensland, from January 1998 to December 1999. There were 1516 children aged 0-14 years who presented following ingestional poisoning. RESULTS: The presentation rates for poisoning were 690, 40 and 67 per 100000 population aged 0-4, 5-9 and 10-14 years, respectively. The admission rates to hospital for poisoning were 144, 14 and 22 per 100000 population aged 0-4, 5-9 and 10-14 years, respectively. Although presentation rates for poisoning were higher in the rural centres the admission rates were disproportionately high for the 0-4 years age group. The agents most frequently ingested were paracetamol, Dimetapp, rodenticides and essential oils. CONCLUSION: There is a need to design and implement interventions aimed at reducing poison exposures and unnecessary hospital admissions in the 0-4 years age group. PMID- 11885706 TI - Maternal awareness of sudden infant death syndrome in North Queensland, Australia: an analysis of infant care practices. AB - OBJECTIVES: To assess awareness of sudden infant death syndrome (SIDS) and risk reducing recommendations in a sample of mothers in North Queensland, Australia, and to examine their infant care practices. METHOD: Interviews conducted with 195 women using a standardized questionnaire between October 1997 and January 1998. RESULTS: 191 questionnaires analyzed; 134 (70.2%) Caucasian and 57 (29.8%) indigenous women. Four women with previous SIDS experience were excluded from the analysis. Eight (4.2%) had never heard of SIDS. Twenty-nine (15.2%) had heard of SIDS and 154 (80.6%) had heard of SIDS and could list risk recommendations to reduce its incidence. Multivariate analysis identified ethnicity as the only significant predictor of maternal knowledge. Indigenous mothers knew less about SIDS: adjusted odds ratio (OR) = 5.4; 95% confidence interval (CI) = [2.1-14.0]. Avoidance of prone sleeping was the most frequently identified recommendation (n = 132), with no smoking in pregnancy (n = 48) and breastfeeding (n = 40) identified least frequently. There were 80.2% of mothers who put their infant in non-prone positions to sleep. Only 48 (25%) women identified smoking in pregnancy, and 93 (48.6%) smoking in the infant's environment as risk factors. Indigenous women were more likely to smoke in their pregnancy (P = 0.004), bed share with their infant (P = 0.0001), and have smokers in the home. CONCLUSION: There is a high level of awareness of SIDS and the main associated risk factor of infant prone sleeping, but the link between SIDS and smoking requires further emphasis. Future campaigns should ensure the SIDS message is delivered more effectively to the indigenous communities. PMID- 11885708 TI - Transcatheter patent ductus arteriosus occlusion: evolution of techniques and results from the 1990s. AB - OBJECTIVE: To review the evolution of transcatheter patent ductus arteriosus (PDA) occlusion techniques and results. METHODS: A single institution, retrospective review including all patients with intention to close a PDA from 1991 to 1998, with no exclusions. RESULTS: Rashkind occluder (n = 65), sideris double-button (n = 6), Cook detachable coil (n = 28) and Amplatzer ductal occluder (n = 4) were used. Successful implantation occurred in 99 of 103 patients. There was a need for a second transcatheter procedure to close residual ductal shunting in 12% of patients: Rashkind umbrellas (n = 8), double-button (n = 1), coils (n = 3). Eight patients (8%) required surgery, including 4 of 6 patients with the double-button occluder. CONCLUSIONS: The Rashkind occluder and the Sideris double-button device both had an unacceptably high rate of residual shunts requiring a second transcatheter procedure or surgical closure. Detachable coils and the Amplatzer ductal occluder have become the current technology of choice for transcatheter PDA closure with high success rates. PMID- 11885709 TI - Growth of infants during the first 18 months of life in urban and rural areas of southern China. AB - OBJECTIVE: Observe the growth pattern of infants from birth to 18 months. METHODOLOGY: A prospective study was conducted from 1994 to 1996 in urban and township-rural areas of Guangdong Province, southern China, recruiting 568 and 257 newborn infants, respectively. Anthropometric data was collected at birth, 1.5, 4, 5, 6, 8, 10, 12, 15, and 18 months. RESULTS: The urban infants at birth had Z-scores of weight for age (WAZ), height for age (HAZ) and weight for height (WHZ) below the National Center for Health Statistics (NCHS) median (P < 0.01). However, from 1.5 to 4 months, the WAZ and WHZ scores were above (P < 0.01), but by 7 months fell and remained at - 0.7 to - 0.8 SD below the NCHS median (P < 0.01). HAZ scores improved after birth, were at the NCHS median to 8 months (P = NS), and then decreased to 0.2-0.4 SD below the NCHS median (P < 0.01). Compared with the urban infants, the township-rural infants were lighter and shorter throughout the first 18 months of life, and from 8 months of age, were - 1 SD or more below the NCHS median. CONCLUSIONS: Growth retardation was found in this sample of Chinese infants. The magnitude of growth retardation was greater for infants in the township-rural area compared with those in the urban area. PMID- 11885710 TI - The effect of breastfeeding on child development at 5 years: a cohort study. AB - OBJECTIVE: It is uncertain to what degree the relationship between breastfeeding and later cognitive development is a true biological effect, or is confounded by psychosocial factors. The study aim was to further investigate this relationship and the effect of duration of breast feeding on cognitive development. METHODS: A total of 3880 children were followed from birth. Breastfeeding duration was measured by questionnaire at 6 months of age and a Peabody Picture Vocabulary Test Revised (PPVT-R) was administered at 5 years. PPVT-R scores were adjusted for the effects of a large array of biological and psychosocial confounders. The relationship between breastfeeding and the mean PPVT-R scores were examined using analysis of variance and multiple linear regression. RESULTS: A strong positive relationship was demonstrated between breastfeeding and the PPVT-R scores with increasing scores with increased duration of breastfeeding. After adjusting for a wide range of biological and social factors, the adjusted mean for those breastfed for 6 months or more was 8.2 points higher for females and 5.8 points for males when compared to those never breastfed. CONCLUSION: These findings suggest a significant benefit to child development is conferred by breastfeeding and is related independently to longer periods of breastfeeding. PMID- 11885711 TI - The sleep and settle questionnaire for parents of infants: psychometric properties. AB - OBJECTIVE: To determine the psychometric properties of a parent-report questionnaire (Sleep and Settle Questionnaire (SSQ)) assessing: (i) the infant's sleep and settling behaviour, and (ii) the parent's level of concern with such behaviours. METHODOLOGY: Test-retest reliability was determined by administering the SSQ to 20 mothers on two occasions, 7-14 days apart. Validity was determined by comparing SSQ responses between mothers with 6-week-old infants who, on a semistructured questionnaire, reported no sleep or settling difficulties (n = 56 60) with those who reported they were experiencing difficulties (n = 133). Further comparison was made with a sample of mothers (n = 34-36) attending a community class on sleep and settling difficulties with infants. Sensitivity to change was determined by comparing mothers' SSQ responses at 6-weeks and 6-months postpartum. RESULTS: The SSQ was found to have low test-retest reliability on items referring to the infants' sleep and settling behaviour, but moderate reliability for the extent that such behaviour bothered the parent. Comparison across the different samples showed good discriminant and concurrent validity. CONCLUSIONS: Parental reports on the SSQ indicates that over a short period (1-2 weeks) the infants' sleep and settling behaviour can change considerably, but that the extent to which such behaviour bothers the parent is more stable. Good validity demonstrates the SSQ is sensitive to differing infant behaviour. It is recommended as both a clinical and research instrument, and could be used to complement assessments focusing on the parent's psychosocial adjustment in the early postpartum period. PMID- 11885712 TI - Specific dangers associated with infants sleeping on sofas. AB - AIM: A study was undertaken to examine specific circumstances that may lead to accidental asphyxial deaths in infants on sofas. METHODS: Coronial files in South Australia (Australia) from 1989 to 1998, and files at the Office of the Medical Examiner in San Diego County (USA) from 1991 to 1998 were searched for all cases of infant deaths occurring on sofas. RESULTS: A total of 10 cases with complete death scene descriptions were found. Four deaths were attributed to sudden infant death syndrome and six deaths to accidental asphyxia, of which four involved shared sleeping with an adult. Lethal circumstances involved infants being overlayed by an adult (n = 2), wedged between an adult and the back of a sofa (n = 1), sleeping with an intoxicated/sedated adult (n = 2), wedged between pillows and the back of a sofa (n = 1), and wedged into the back of a sofa (n = 1). CONCLUSIONS: Although shared sleeping of an adult with an infant on a sofa may result in accidental asphyxia, there is also the potential for wedging and accidental asphyxia of infants sleeping alone on a sofa. For this reason the use of sofas for both shared and solitary infant sleeping is discouraged. PMID- 11885713 TI - Successful clot lysis using low dose of streptokinase in 22 neonates with aortic thromboses. AB - OBJECTIVE: To determine whether intravenous infusion of low dose of streptokinase was effective in lysing umbilical arterial catheter (UAC)-associated aortic thrombi. METHOD: A prospective cohort study of 31 consecutive newborn infants with UAC-associated aortic thrombi which were detected by abdominal ultrasonography after removal of UAC. Twenty-two infants were treated with intravenous infusion of low dose (1000 U/h) streptokinase, while nine others were not treated due to various contra-indications. Thrombolysis occurred after a mean interval of 2.2 days (standard deviation (SD) = 1.8) in the treated infants. In the untreated infants, spontaneous thrombolysis occurred significantly later, after a mean interval of 16.9 days (SD = 14.7) (95% confidence intervals of difference between mean intervals - 26.0, - 3.4; P = 0.02). Only one treated infant developed mild bleeding directly attributed to streptokinase therapy. CONCLUSION: Low dose streptokinase infusion was effective and safe in thrombolysing UAC-associated aortic thrombi. PMID- 11885714 TI - Ten years' experience of persistent hyperinsulinaemic hypoglycaemia of infancy. AB - OBJECTIVE: To review the presentation, management and outcome of persistent hyperinsulinaemic hypoglycaemia of infancy seen at the Royal Alexandra Hospital for Children over a 10 year period. METHODOLOGY: A retrospective review of 20 subjects was performed. As well as laboratory data, data were collected on clinical presentation, medical and surgical management and developmental outcome. RESULTS: Twenty subjects (11 male) were identified with presentation at a median age of 1.5 months (range 0-10 months), with 10 (50%) presenting in the first week of life. Only 20% of patients were large for gestational age. Diagnosis was made on the basis of high glucose requirements and inappropriately high insulin levels at the time of hypoglycaemia. Eight (40%) responded well to diazoxide treatment alone, seven (35%) received diazoxide in combination with other short-term medical therapy initially and five (25%) required pancreatectomy (repeat surgery in three). Those who required surgery had a higher mean birth weight. Infants presenting in the first week of life were less likely to respond to diazoxide. At the time of last review, eight (40%) of those treated medically had ceased all treatment. Two of the five cases requiring pancreatectomy now require insulin treatment. Neurodevelopmental assessment was normal in 11 (55%), mild delay was found in six (30%) and moderate or severe delay was found in three (15%). CONCLUSIONS: Persistent hyperinsulinaemic hypoglycaemia of infancy remains a major diagnostic and management challenge. Early suspicion and recognition is critical with definitive investigation and medical therapy to avoid hypoglycaemia, with pancreatectomy in medically unresponsive cases. Normal neurodevelopmental outcome was found in only 55% of cases. PMID- 11885715 TI - Inspired gas humidity during mechanical ventilation: effects of humidification chamber, airway temperature probe position and environmental conditions. AB - OBJECTIVES: To determine the inspired gas humidity during mechanical ventilation with: (i) four different humidification chambers; (ii) two airway temperature probe (ATP) positions; (iii) five different humidicrib temperatures; and (iv) insulating the inspiratory limb with bubble wrap. METHODOLOGY: An observational study in the Neonatal Laboratory and Neonatal Intensive Care Unit, Westmead Hospital. The humidity of the inspired gas was measured at the proximal end of the endotracheal tube (ETT) during mechanical ventilation. Inspired humidity measurements were made with four different humidification chambers (Fisher & Paykel (F&P Healthcare Pty Ltd, Auckland, New Zealand) auto refill MR290. F&P manual refill MR310, Suruga (Suruga Inc. Humidifiers, Vincent Medical, Dongguan, China) manual refill MI-20 and MI-10F) with the humidity control (relative humidity setting) set at - 2. Measurements were made with the ATP positioned either; (A) at the distal end of the inspiratory tube inside the humidicrib or (B) outside the humidicrib 50 cm proximal to the ETT. The inspired gas temperatures were set at 36.5 degrees C and at 39.0 degrees C, respectively. For each of the different humidification chambers and ATP positions, inspired humidity measurements were made with the humidicrib temperature set at 30.8, 32.9, 35.2, 36.2, or 37.2 degrees C. Two further sets of measurements were made, one with the inspiratory limb insulated with bubble wrap and the second set without bubble wrap. RESULTS: There were significant differences in inspired humidity with the four humidification chambers at both ATP positions at all humidicrib temperatures. Both Suruga humidification chambers produced significantly higher inspired gas humidities under most conditions. Positioning the ATP outside the humidicrib produced significantly higher inspired gas humidities than with the ATP inside the humidicrib. Insulating the inspiratory tubing with bubble wrap also significantly improved the inspired gas humidity. CONCLUSIONS: Significant differences in inspired gas humidity were found with the humidification chambers tested. The position of the ATP and the set temperature had a significant impact on the absolute humidity of the inspired gas. In general, higher inspired gas humidities were obtained with the ATP outside the humidicrib. However, condensation of water close to the ETT appeared at low humidicrib temperatures (< 36.2 degrees C) with the ATP outside the humidicrib and extreme care should be taken that particulate water does not enter the lungs under these conditions. PMID- 11885716 TI - Inspired gas temperature during mechanical ventilation: effects of environmental temperature and airway temperature probe position. AB - OBJECTIVES: To determine the inspired gas temperature during mechanical ventilation with: (i) five different humidicrib temperatures; (ii) two airway temperature probe (ATP) positions; and (iii) four ATP adaptors. METHODOLOGY: An observational study in the Neonatal Intensive Care Laboratory, Westmead Hospital. The inspired gas temperature was measured at the proximal end of the endotracheal tube (ETT) during conventional mechanical ventilation using a Tele-thermometer. Inspired gas temperature measurements were made with: (i) the humidicrib temperature set at 30.8. 32.9, 35.2, 36.2. or 37.2 degrees C; (ii) the ATP either (A) positioned inside the humidicrib at the distal end of the inspiratory tubing or (B) positioned outside the humidicrib 50 cm proximal to the ETT, with the inspired gas temperatures set at 36.5 and 39.0 degrees C, respectively; and (iii) the measurements repeated with four different ATP adaptors at each humidicrib temperature and each ATP position. RESULTS: With the ATP inside the humidicrib, there were no significant differences between set and actual inspired gas temperature. However, with the ATP outside the humidicrib, there were significant decreases in inspired gas temperature at each humidicrib temperature. For instance, with the ATP outside the humidicrib and set at 39.0 degrees C, the inspired gas temperature decreased to 34.7+/-0.2 degrees C at a humidicrib temperature of 30.8 degrees C and to 37.7+/-0.2 degrees C at a humidicrib temperature of 37.2 degrees C. The type of ATP adaptor also had a significant effect on inspired gas temperature. CONCLUSIONS: With the ATP placed outside the humidicrib and with variations of humidicrib temperature, infants are likely to have inspired gas temperatures that are significantly different to the desired temperature. Certain ATP adaptors cause these variations in inspired gas temperature to be more pronounced. Extreme care must be used to avoid suboptimal inspired gas temperatures with these environmental variations and the ATP positioned outside the humidicrib. PMID- 11885717 TI - Parent-requested treatment. AB - The decision about EPO was referred to and made by the Drug Committee, a committee of physicians, nurses and pharmacists. This committee has perforce to make decisions about drugs and vaccines, decisions which sometimes have a significant ethical component due to concerns about cost, safety and efficacy. Our hospital is considering developing a Clinical Ethics Advisory Committee, to assist with difficult ethical decisions such as this one. Should such a committee be asked to make acute ethical judgements on patient management? Larcher describes his ideal Hospital Ethics Committee as nonprescriptive, and suggests a more appropriate role is retrospective analysis and reflective discussion of clinical ethical problems. Such discussion may help with future rather than current management issues, and can help support clinicians in their decisions and hospital staff in their management of patients. PMID- 11885718 TI - Bones, groans and blasts. AB - A 14-year-old girl presented with the acute onset of gastrointestinal symptoms due to hypercalcaemia. Chest X-ray revealed osteolytic lesions in the ribs which in conjunction with a normal parathyroid hormone level raised the possibility of malignancy. Despite the absence of blast cells in her blood film, the bone marrow biopsy was diagnostic of acute lymphoblastic leukaemia. She responded well to treatment with pamidronate and chemotherapy. PMID- 11885719 TI - Fevers and mouth ulcers. AB - Mouth ulcers are commonly caused by infection but may be due to neutropenia. The most common form of hyper-IgM syndrome is of X-linked inheritance and caused by CD40 ligand gene mutations. Consider hyper-IgM syndrome in a male child with recurrent bacterial or opportunistic infections, neutropenia, hypogammaglobulinaemia (IgG and IgA) and normal T- and B-cell counts. In X-linked hyper-IgM syndrome: - the serum IgM concentration is normal in about 50% of cases. - transient or persistent neutropenia occurs in 70% of cases. First-line therapeutic options for hyper-IgM syndrome include regular intravenous immunoglobulin and prophylactic trimethoprimsulphamethoxazole. PMID- 11885720 TI - Carbohydrate-deficient glycoprotein syndrome 1b: a new answer to an old diagnostic dilemma. AB - A patient with carbohydrate-deficient glycoprotein syndrome type 1b (CDGS1b) is reported. The patient presented at 5 months of age with failure to thrive, prolonged diarrhoea, hepatomegaly and elevated serum liver transaminases. Liver biopsy showed steatosis. A low serum albumin and elevated serum liver transaminases persisted throughout childhood during which he had repeated infectious illnesses. From the age of 10 years he had oesophageal and duodenal ulceration together with recurrent bacterial cholangitis. Liver biopsy demonstrated hepatic fibrosis. CDGS1b was suspected, supported by the finding of a protein-losing enteropathy and finally confirmed by showing a reduced phosphomannoseisomerase activity. This case illustrates a rare condition with a wide range of presentations. PMID- 11885721 TI - The diagnosis and management of neonatal urinary ascites. AB - Urinary ascites in a newborn infant is unusual and most commonly indicates a disruption to the integrity of the urinary tract. The following report describes a case of urinary ascites, probably due to bladder rupture caused by umbilical artery catheterization, associated with hyponatremia, hyperkalemia and elevated serum creatinine. This unusual biochemical profile is characteristic of urinary 'autodialysis' and was corrected by bladder drainage. PMID- 11885722 TI - 3beta-hydroxy-delta5 -C27-steroid dehydrogenase deficiency: diagnosis and treatment. AB - The aim of this study was to evaluate the effects of bile acid treatment and to obtain further information about the pathway of bile acid biosynthesis in a patient with 3beta-hydroxy-delta5-C27-steroid dehydrogenase/isomerase (3beta-HSD) deficiency by gas chromatography-mass spectrometry. Results showed that at 2 months of age, 3beta-hydroxy-5-cholen-24-oic acid (3.0 micromol/mmol Cr, 7.9%) was detected in the urine in essentially the same relative amount as 3beta,7alpha dihydroxy- and 3beta,7alpha,12alpha-trihydroxy-5-cholen-24-oic acids (3.7 micromol/mmol Cr, 9.8%) during ursodeoxycholic acid treatment combined with prednisolone. As a result, diagnosis was delayed until 18 months of age. One month later with substitution of chenodeoxycholic acid treatment, urinary 3beta,7alpha-dihydroxy- and 3beta,7alpha,12alpha-trihydroxy-5-cholen-24-oic acids decreased significantly, and subsequent improvement of liver dysfunction was accelerated. Chenodeoxycholic acid treatment is useful in 3beta-HSD deficiency. However, in the diagnosis of this disease in early life, it should be noted that the acidic pathway may be the major route for bile acid biosynthesis in the neonatal period. Diagnosis of 3beta-HSD deficiency may have been delayed by administration of ursodeoxycholic acid, resulting in prolonged diagnostic investigation in this child with cholestasis. Further, use of prednisolone may have been contraindicated. PMID- 11885723 TI - Multiple fractures in a 3-month-old infant with severe infantile osteopetrosis. AB - A diagnosis of severe infantile, autosomal recessive osteopetrosis (I-ARO) was made in a 3-month-old female based on characteristic radiological and histological findings. The finding of multiple fractures at presentation in this infant is highly unusual. Deficiency of carbonic-anhydrase type II was excluded. PMID- 11885724 TI - Use of acridine orange leukocyte cytospin test in diagnosis of neonatal sepsis. PMID- 11885725 TI - Asymptomatic oesophageal perforation in a neonate. PMID- 11885726 TI - Upper airway obstruction induced by a caustic substance found responsive to nebulised adrenaline. PMID- 11885727 TI - Use of intravenous megadose corticosteroid in a child with thrombocytopenic purpura due to mumps. PMID- 11885728 TI - Delayed diagnosis and treatment of umbilical arterial catheter-associated aortic thrombus. PMID- 11885729 TI - Percutaneous intravenous central catheters. PMID- 11885730 TI - Meningococcal vaccines: advances but new questions? PMID- 11885731 TI - A 10-year serogroup B meningococcal disease epidemic in New Zealand: descriptive epidemiology, 1991-2000. AB - OBJECTIVE: New Zealand has experienced an epidemic of meningococcal disease since 1991. This paper describes the characteristics of this epidemic during its first 10 years (1991-2000), current control measures, and potential future interventions. METHODOLOGY: Meningococcal disease surveillance in New Zealand uses combined notification and laboratory data. Population census data from 1991 and 1996 were used to calculate disease rates. RESULTS: The annual incidence of meningococcal disease increased from 53 cases (1.6 per 100 000 population) in the pre-epidemic year of 1990 to a peak of 613 (16.9 per 100000) in 1997, followed by consistently raised rates. Over the 1996-2000 period, there was an average of 502 cases per year (13.9 per 100 000). The epidemic has resulted in 3547 cases since 1991 approximately 3000 in excess of the number expected based on pre-epidemic disease incidence. Of the total cases, 158 (4.5%) were fatal. A disproportionately large number of cases have been in Maori and Pacific Islands children in the northern part of the North Island of New Zealand. Since 1991, the epidemic has increasingly been dominated by serogroup B meningococci with subtype P1.7b,4, which by 2000 accounted for 84.6% of all cases for whom this testing was carried out. The majority of these organisms were characterised as B:4:P1.7b,4. CONCLUSION: Meningococcal disease rates are likely to remain elevated in New Zealand for at least several more years. A vaccine which could induce immunity to the P1.7b,4 PorA subtype may have a role in controlling this epidemic. Efforts are underway to obtain and trial such a vaccine. Measures are also underway to reduce overcrowded living conditions which are contributing to the epidemic. Early recognition and antibiotic treatment of cases improves outcomes and should continue to be promoted. Integrated notification and laboratory-based surveillance of meningococcal disease provides relatively complete surveillance of this disease in New Zealand and has supported the development of public health interventions. PMID- 11885732 TI - Meningococcal disease and vaccination in North America. AB - In North America, meningococcal disease occurs at a rate of I case per 100000 population per year, producing 2725 cases notified in the US in 1998 and 155 laboratory confirmed cases in Canada in the same year. A majority of these cases occur in the winter season and in early childhood, with a case fatality rate of approximately 10%. There has been an increase in the proportion of cases due to serogroup Y meningococci over the past decade: in 1995-98 in the US, 33% of cases were due to serogroup B, 28% were due to serogroup C and 34% were due to serogroup Y; in Canada in 1995-96, 47% of cases were due to serogroup B, 40% were due to serogroup C and 10% were due to serogroup Y. Outbreaks due to serogroup C were more common in the 1990s in the US and a number of outbreaks have occurred in Canada due to organisms from the hypervirulent ET-37 complex. College freshmen in the US in dormitories were found to be at an increased risk of disease. In addition, over the past 10 years, an outbreak of serogroup B disease occurred in the Pacific North-west of the US, with a fourfold increased rate of disease in that region. The explanations for these changes in epidemiology are unknown, but probably reflect the appearance of hypervirulent clones of meningococci and/or changing levels of population immunity. Meningococcal serogroup C polysaccharide protein conjugate vaccines have been introduced recently in the UK, seem efficacious and offer the potential to reduce the burden of disease in the US and Canada too. Because serogroups other than serogroup C are prevalent in North America, a combination polysaccharide-protein vaccine, including C, Y and possibly W135 serogroups, would be attractive for this population. Although not currently an issue in industrialized nations, inclusion of serogroup A conjugates in any future vaccine policy would be an important decision in driving global prevention of meningococcal disease. A meningococcal conjugate vaccine against serogroup C was licensed in Canada in April 2001. PMID- 11885733 TI - Meningococcal vaccination for adolescents? An economic evaluation in Victoria. AB - OBJECTIVE: To undertake an economic evaluation of the options for vaccination of adolescents using meningococcal polysaccharide vaccine based on Victorian data. METHODOLOGY: Cost-effectiveness and cost-benefit analyses of three options for vaccination were undertaken for hypothetical populations aged 15-19 years. Baseline analyses assumed a single year of programme implementation and vaccine protection of 5 years. Sensitivity analyses of key variables were performed. Outcomes included the number of people vaccinated, cases averted, life years saved and disability adjusted life years (DALY) averted. Lost earnings avoided were included as a measure of vaccination benefit in cost-benefit analyses. RESULTS: Vaccination of people in Years 10-12 (secondary school) and first year university within a defined population with a high rate of disease was the most cost-effective option. Excluding direct cost savings and compared with no vaccination, this resulted in a discounted cost per DALY avoided of $17646 and benefits exceeding costs in discounted terms. The 'break-even' incidence rate for this option in the cost-benefit analysis was 11.9/100000. CONCLUSIONS: Economic evidence favours the use of vaccination within well-defined populations with a high rate of disease. PMID- 11885734 TI - Epidemiology of meningococcal disease in Australia. AB - A review of the epidemiology of meningococcal disease (MD) in Australia was undertaken, with particular emphasis on the 1990s, when national strain differentiation data became available. The data included a review of clinical and laboratory notification data and published reports on clusters and outbreaks. There have been considerable changes in the patterns of MD in the 1990s. In some cases, these changes can be related to the dominance of a particular phenotype. In the early 1990s, widely scattered urban and rural clusters were associated with the phenotype C:2b:P1.2 and strains were closely genetically related. Larger urban clusters and increased numbers of cases in adolescents and young adults were most obvious in New South Wales in the mid-1990s and were associated with a phenotype C:2a:P1.5. This ET-15 clone of the ET-37 complex caused similar patterns of MD to those seen in other countries as part of the global spread of the clone. In contrast, the B:4:P1.4 phenotype, with close genetic similarities to New Zealand strains, did not cause the hyperendemic disease seen in New Zealand this decade. The epidemiology of MD will continue to exhibit considerable variation due, at least in part, to the genetic flexibility of meningococci. Information about strain variation expands our understanding of changing patterns of disease. PMID- 11885735 TI - Introduction of a conjugate meningococcal type C vaccine programme in the UK. AB - OBJECTIVE: The purpose of the UK initiative was to accelerate the development and introduction of new conjugate meningococcal C vaccine into the routine immunisation programme, and to implement a catch-up campaign based on the disease epidemiology. METHODOLOGY: Collaboration between Government supported institutions and the vaccine industry lead to a collaborative programme of research designed to answer policy specific questions and to accelerate the availability of new vaccines. RESULTS: Three new conjugate meningococcal C vaccines were developed and licensed for use in the UK after satisfactory data on safety and immunogenicity had been generated. A nationwide campaign was designed to offer vaccine to all infants at the same time as their three doses of primary immunisations, two doses were offered to children over 4 months and under 1 year old; all those over 1 and under 18 years old were offered a single dose of vaccine. The programme was on course for completion within approximately twelve months, with around 15 million immunisations being offered. The programme was implemented simultaneously through school and primary care services. CONCLUSIONS: A safe and effective new vaccine, against group C meningococcal disease, has been introduced into the UK immunisation programme after just a 5 year development to implementation process. Early indications point to high coverage and impacts on disease are already apparent in the groups that have been immunised. These vaccines may play an equally important role in other countries where there is a significant burden from Group C meningococcal disease. PMID- 11885736 TI - Laboratory enhanced surveillance for meningococcal disease in Victoria. AB - OBJECTIVE: To describe the epidemiological and microbiological characteristics and notification patterns of invasive meningococcal disease (IMD) in Victoria between 1990 and 1999. METHODS: Cases of IMD occurring between 1990 and 1995 identified in any of three databases were combined, matching where possible. Statistical modelling provided estimates of cases missing from all datasets. Notification sources for 1999 and 2000 cases were identified. Cases identified from notification and laboratory results provided the data to describe IMD epidemiology between 1990 and 1999. RESULTS: Between 1990 and 1995, 479 cases of IMD were identified. Three individual datasets each identified between 62 and 82% of cases and 47% of cases were identified in all three datasets. Statistical modelling estimated that between 37 and 83 additional cases were not identified by any dataset. Serogroup B and C strains caused 63 and 33% of culture-positive cases, respectively, with a substantial rise in serogroup C cases in 1999. Epidemiological characteristics remained relatively constant between 1990 and 1998, but an increase in patient age was seen in cases with serogroup C disease in 1999. In addition to three clonal strains seen elsewhere, an additional strain was identified that was unique to Victoria. Since January 1999, only 72% of notifications have come from treating doctors. CONCLUSIONS: Meningococcal disease is of increasing public health significance in Victoria. Laboratory enhanced notification has improved case identification and detailed microbiological information has improved our understanding of the changing epidemiology of this disease. Collaboration with laboratories and other agencies, active investigation of putative cases and microbiological monitoring are important elements in supporting public health decisions about the control of IMD. PMID- 11885737 TI - Influence of sample preparation, staining procedure and analysis conditions on bull sperm head morphometry using the morphology analyser integrated visual optical system. AB - The importance of standardizing the procedures of sample and slide preparation for computer-assisted morphologic analysis has been emphasized in human and veterinary andrology. The purpose of this study was to optimize slide preparation (dilution grade and sperm washing), staining procedures and analysis conditions (colour of light source and objective magnification) for the morphometric analysis of bull spermatozoa using the Hamilton Thorne morphology analyzer integrated visual optical system (IVOS). For experiment 1, one ejaculate was collected from one bull and diluted to 200,000-300,000 spermatozoa/microl. Slides were prepared and stained using seven different procedures: rapid Papanicolaou (PAP), rapid Papanicolaou with prolonged staining times (PAP+). Diff-Quik (DIF), haematoxylin (HEM). Farelly (FAR), Spermac (SPER) and the modified GZIN (MGZIN) staining. All slides were analysed using a Hamilton Thorne Morphology Analyser IVOS equipped alternatively with a red, green or blue light source, and a 40x or 100x oil immersion objective. Recognition and digitization errors as well as morphometric parameters were determined. The IVOS was unable to detect DIF stained spermatozoa. The GZIN and the SPER staining as well as the blue light source led to unsatisfactory results. Among the staining methods examined, the FAR, HEM, PAP+, and PAP staining, preferably in combination with the green light source, and the 40x objective yielded optimal results concerning sperm recognition and digitization. The 100x objective did not allow reliable analysis of the sperm heads because of a frequently appearing digitization error. For experiment 2, three ejaculates were collected from each of three bulls and diluted to five dilution grades (100 000-500 000 spermatozoa/microl). An aliquot of each dilution grade was washed additionally. The percentage of correctly digitized sperm heads decreased with increasing spermatozoal concentration. However, the evaluation speed increased. The range of 200 000-300 000 spermatozoa/microl appeared to be a reasonable compromise for both criteria. Sperm washing failed to further improve the analysis results. Sperm head dimensions were influenced significantly by all variations of the methods in both experiments. In conclusion, using the proposed methods, the IVOS allows precise and reliable morphometric analyses of bull spermatozoa. The consistent application of these procedures may lead to an inter-laboratory standardization and to further establishment of generally accepted morphometric criteria used in human andrology (e.g. World Health Organisation or strict criteria). PMID- 11885738 TI - First identification of caldesmon transcripts in bovine oviduct epithelial cells in vitro by means of an RNA differential display technique examining culture induced expression changes. AB - Innovative molecular biology techniques enable the quick evaluation of distinct gene expression pattern of cells or tissues. Hitherto, a cell-type-specific behaviour has been difficult to evaluate. In addition to standard morphological and immunological criteria in this study the expression of in vitro-cultured bovine oviduct epithelial cells (BOECs) was compared with fresh cells by RNA arbitrarily primed polymerase chain reaction (RAP-PCR). The cultured cells showed mitotic activity during the whole culture period (6 days) until they had reached about 80% confluency. Remarkable results of a random transcript screening using fresh versus cultured BOECs were reported, in which a single PCR product appeared during culture, but was absent in fresh cells. Sequence analysis of the culture induced 522 bp fragment revealed a high homology (87%) to caldesmon (CaD) of various species and a 92% homology to a short cDNA fragment of a bovine non muscle CaD. Specific, cross-species PCR primers were used to elongate this partial sequence (1,036 bp). This resulting cDNA showed an open reading frame and was identified as a bovine non-muscle CaD isoform. When compared with human non muscle CaDs (89% homology) a deletion of 2 codons was observed. According to sequential culture experiments, CaD expression was not found in fresh BOECs but specific transcripts appeared within 48-113 h under specific culture conditions. It is likely that augmented CaD expression in cultured BOECs may reflect the cell effort adapting to specific culture conditions. The hypothesis that increased CaD levels could be important to facilitate adherence and spreading by formation of new stress fibres can be favoured. This first identification of caldesmon expression being specifically induced during in vitro culture demonstrates the potential of RAP-PCR for the analysis and validation of cell culture techniques. PMID- 11885739 TI - Colour-coded and pulsed Doppler sonography of the canine testis, epididymis and prostate gland: physiological and pathological findings. AB - Two-dimensional ultrasound was used in combination with colour-coded and pulsed Doppler sonography to study the blood flow of the testes and prostate gland in a total of 30 male dogs. After detection of the vessels by colour-coded Doppler sonography, the blood flow patterns were determined by pulsed Doppler sonography measuring and describing the systolic and diastolic peak velocity (SPV, DPV), the end-diastolic velocity, the time-averaged maximum velocity (TAMAX), the pulsatility and resistance index, as well as the ratios of the systolic peak velocity and end-diastolic velocity and of the systolic and diastolic peak velocities. The blood flow of the testicular artery was measured within the pampiniform plexus and the marginal location. The prostatic blood supply was measured in the artery of the deferential duct (cranial), the prostatic artery outside (lateral) and within the gland (subcapsular). The physiological testicular flow pattern was monophasic with a high diastolic flow. Testes with neoplastic alterations showed a significant increase of SPV and TAMAX. The epididymal vessels could not be detected. Under physiological conditions the prostatic blood flow pattern was biphasic in the cranial and lateral location and monophasic in the subcapsular location. Benign prostatic hyperplasia was characterized by a significant increase of SPV, DPV and TAMAX. The results of the present investigation demonstrate that the colour-coded and pulsed Doppler sonography give additional valuable information which improves the andrological diagnostics in the dog. PMID- 11885740 TI - Semen collection, examination and spermiogram in ostriches. AB - The level of fertility in the male ostrich exerts considerable influence on the efficiency of the fertilization procedure, and thus also on reproductive performance. The determination of the reproductive capacity is of particular interest with regard to the selection of single individuals for optimizing reproduction ratios. Although the breeding and raising of ostriches has become increasingly important in many countries, little research has been completed on reproductive parameters and factors that may possibly influence them. This study presents observations made concerning the quantity and quality of sperm as found in the spermatological testing of 411 ejaculate samples taken from male ostriches on two farms in Namibia. The semen volume varied between 0.1 and 1.5 ml (mean, 0.64 ml). Normal ejaculate colours ranged from white to ivory; the consistency ranged from thin creamy to viscous. The measured pH values lay between 6.4 and 8.0 (mean, 7.3). Microscopic investigations revealed sperm concentrations of 8.9 78.1 million/microl and individual sperm motility from 42 to 96% (mean, 78%). No mass motility was detectable in 42% of the ejaculates; weak mass motility was found in 46%, and clear mass movements were to be found in only 12% of samples. Regarding the morphology of the sperm, 5 to 26% were abnormal (mean, 17%) and 4 to 28% (mean, 20%) were dead. Seasonal patterns of sperm concentration and the influence of frequency of semen collection were investigated in a group of 56 healthy male ostriches. Peak sperm concentrations were found at the beginning of the breeding season in spring; the lowest values were found at the end of the breeding season in autumn. The highest quality ejaculate was obtained from those males whose semen was collected once a week. The results of this study provide fundamental data for the establishment of minimum quality requirements for ostrich sperm to be met by individuals receiving certification as breeding animals and for the selection of suitable males for use in artificial insemination. PMID- 11885741 TI - Combined gnRH and PGF2alpha application in cows with endometritis puerperalis treated with antibiotics. AB - The investigations were carried out on a total of 70 cows with puerperal endometritis. In addition to intrauterine antibiotic treatment, 30 experimental animals were administered 20 microg GnRH analogue, buserelin, between days 10 and 12 post-partum followed by 500 microg PGF2alpha analogue, cloprostenol, 10 days later. Forty control cows were treated only with intrauterine antibiotics. Blood samples for progesterone determination were collected from the tail vein twice weekly until day 70 post-partum. The first rise in progesterone level above 3.18 nmol/l occurred significantly earlier in the experimental than in control cows (21.6 +/- 9.2 versus 27.8 +/- 12.3 days; p < or = 0.05). The duration of the first cycle post-partum was 15.0 +/- 4.3 days in experimental and 19.7 +/- 7.3 days in control animals (p < or = 0.05). However, no significant differences were observed in the occurrence of first oestrus post-partum. The involution of the uterus was improved after hormone treatment. At day 42 post-partum, completion of uterine involution was found in 93.3% of hormone-treated cows and in 82.5% of those treated with antibiotic only (p < or = 0.05). Clinical recovery was 96.6% in the experimental and 82.5% in the control group (p < or = 0.05). First service pregnancy rate was significantly better in hormone-treated than control cows (51.7 versus 36.4%; p < or = 0.05). Total pregnancy rate and insemination index values were not significantly improved following GnRH and PGF2alpha treatment. The average service period was 89.8 +/- 21.2 days in cows after hormone treatment, and 112.6 +/- 24.5 days in control cows. The difference was statistically significant (p < or = 0.05). These results indicate, that the sequential GnRH and PGF2alpha application in cows with puerperal endometritis positively affected ovarian function and uterine involution, resulting in improved fertility performance. PMID- 11885742 TI - Clinical and bacteriological aspects on the use of oxytetracycline and flunixin in primiparous cows with induced retained placenta and post-partal endometritis. AB - Retention of the fetal membranes and post-partal endometritis (RFM) are common problems in dairy cows. Treatment often includes manual removal of the placenta in combination with antibiotic treatment. Earlier studies have shown that cows with endometritis post-partum have a strong tendency to recover spontaneously. The present study focused on treatments of post-partal endometritis with the prostaglandin synthesis inhibitor, flunixin (F) either alone or combined with oxytetracycline (T). The study was conducted in two experiments, using 12 primiparous cows in each. As a model for RFM, premature parturition was induced in late pregnant heifers by injecting PGF2alpha (25 mg i.m.) twice with a 24 h interval. In each experiment the cows were set into four groups and treated with either T (10 mg/kg BW i.m. once daily), F (2.2 mg/kg BW p.o. twice daily), a combination of T and F (dosage, as above) or conservatively (group 0, no drugs). The treatment periods lasted from days 11-14 post-partum in experiment I (groups T1, F1, TF1 and 0) and from days 3-6 post-partum in experiment 2 (groups T2, F2, TF2 and 0). Jugular vein blood samples were collected for analyses of flunixin and total white blood cells. Uterine biopsies were collected twice weekly for investigation of endometrial microbiology. Rectal palpation and ultrasonographic examinations were performed three times weekly for investigations of uterine involution and ovarian activity. No attempts were made to remove the placentas manually. The experiment lasted until day 56 post-partum. The induction of parturition was successful in all heifers and 22 of 24 animals had RFM. All RFM cows had bacterial endometritis. The predominant bacteria were Escherichia coli alpha-haemolytic streptococci, Fusobacterium necrophorum, Arcanobacterium (Actinomyces) pyogenes, Bacteroides spp., Pasteurella spp. and Proteus spp. Fusobacterium necrophorum and A. pyogenes could be isolated for 3-5 weeks post partum and E. coli Pasteurella and Proteus could be isolated for 2-3 weeks post partum. Animals treated with tetracycline after placental shedding (T1 and TF1) had a more rapid recovery from infections with A. pyogenes and F. necrophorum than animals that were not treated with tetracycline. No other genera were affected. Antibiotic treatment before placental shedding (T2 and TF2) did not shorten the uterine infection but altered the bacterial flora, seen as an overgrowth of Proteus spp. (p < 0.05) and increased frequency of Pasteurella (p < 0.05). The alpha-haemolytic streptococci were less common in T2 and TF2 than in other groups (NS). Antibiotic treatment of cows before placental shedding (T2 or TF2, n = 6) postponed detachment of placenta compared to cows were no antibiotics were administered before placental shedding (T1, TF1, F1, F2 and 0, n = 16. 9.8 days pp (median) versus p = 0.004). Neither treatment shortened uterine involution. Flunixin treatments did not seem to influence recovery from infection or uterine involution. It was concluded that early oxytetracycline treatment of retained fetal membranes in the cow did not shorten the uterine involution or uterine infection but it did slow down the detachment process of the retained placenta. Oxytetracycline treatment after placental shedding might shorten the uterine infection but otherwise did not affect the clinical results. Flunixin treatment had no influence on the clinical outcome of the disease. PMID- 11885743 TI - Effectiveness of cabergoline for termination of pregnancy in silver fox (Vulpes vulpes fulva). AB - Red foxes (Vulpes vulpes) are a major pest species in Europe and Australia. Traditional methods of control such as hunting or poisoning are no longer sufficient or feasible. As with domestic dogs and cats, prolactin (PRL) in the vixen is an essential luteotropin during the second half of gestation. Hence, PRL inhibitors such as cabergoline have been used to induce abortions. Eighteen mated silver fox vixens (three groups of six foxes each) were treated orally with a placebo of paraffin oil (I), or with 15 microg/kg cabergoline in feed once (11) or twice (III), on day 30 (I and II) or days 30 and 32 (III) post-coitum. Blood samples were taken prior to and after treatments and concentrations of PRL and progesterone (P4) were determined. Normal parturitions were observed in five of six, five of six and two of six vixens in groups I, II and III, respectively. In group III plasma concentrations of PRL and P4 decreased significantly but only temporarily. This drop in hormone concentrations was more pronounced in the vixens that did not carry to term. In conclusion, doses in excess of 15 microg/kg of cabergoline are likely to prevent the development of fetuses to term in pregnant vixens. PMID- 11885744 TI - Neuroendocrine regulation of growth hormone and luteinizing hormone in fetal and neonatal pig. AB - Ontogeny of the production and the regulation of growth hormone and Luteinizing hormone is studied in a series of experiments utilizing male and female pigs at different fetal and neonatal stages. Growth hormone mRNA is detectable in both sexes as early as d. 50 p.c. The mRNA levels increase to reach the maximum levels at d. 95-110 p.c. Plasma levels of GH follow the developmental patterns of GH mRNA. A sex difference is evident around d. 80-90 p.c. with males having higher GH levels than females. The stimulatory but not the inhibitory mechanisms of GH secretion are fully functioning in the pig fetus. LHbeta mRNA is detectable earlier in females (d. 50 p.c.) than in males (d. 65 p.c.). Plasma concentrations of LH increase with fetal age in female fetuses, but in male fetuses there is no distinct developmental pattern evident. Basal LH secretion achieves maximum levels in both sexes after birth. Opioids do modulate fetal LH secretion, however, the mode of their action is age-dependent. PMID- 11885745 TI - Uterine responses to exogenous oxytocin before and after pre-partum luteolysis in the cow. AB - The aim of this study was to test the functional status of uterine oxytocin receptors in cows in vivo around parturition. The animals received consecutive, intra-arterial injections of 800, 1,600 and 3,200 mU of oxytocin at three different stages: during late gestation (days 260-274), at 12 h and at 24 h after intramuscular injection of a prostaglandin F2alpha analogue at day 275 to induce parturition. Cows (n = 6) had been provided with myometrial electrodes and a catheter had been installed in the aorta and in a branch of the uterine vein (UV). Regular blood samples were obtained from the UV from 5 min before until 45 min after each oxytocin injection to measure plasma levels of prostaglandin F2alpha (PGF2alpha) and oxytocin. Uterine electromyographic (EMG) activity was registered continuously during each experiment. The increase of oxytocin levels in UV plasma after intra-arterial injections was dose dependent (p < 0.02). Pre- and post treatment oxytocin levels at 24 h after induction of parturition were significantly increased (p = 0.0313). Both during late pregnancy and at 12 h after induction of parturition, oxytocin caused a significant increase in EMG activity (p = 0.022). After the 3,200 mU dose the increase was significantly higher than with the other 2 doses (p = 0.004). After each dose, EMG activity returned to baseline levels within some 15 min. At 24 h after induction of parturition, the pre-treatment level of EMG activity had increased. Doses of 800 mU and 1,600 mU of oxytocin produced a significant (p = 0.022) increment of EMG activity, which was of the same magnitude as during the preceding stages; after 3,200 mU of oxytocin the response was significantly higher than before (p = 0.008). No significant increases of PGF2alpha levels in UV plasma could be measured after oxytocin injections at any of the three stages. It is concluded that the myometrium of the pregnant cow responds in vivo to physiological doses of oxytocin. At 24 h after induction of parturition, when luteolysis has occurred and a parturient pattern of parturient myometrial activity has already started to develop, the response is enhanced. Physiological doses of oxytocin did not evoke a spurt release of PGF2alpha in uterine venous blood during the peripartal period. PMID- 11885746 TI - Faecal oestrogens and progesterone metabolites in mares of different breeds during the last trimester of pregnancy. AB - Non-invasive pregnancy diagnosis in mares by measuring faecal oestrogens has been performed over years with great accuracy. However, results have indicated breed related differences in the amount of excreted steroids during late pregnancy. Therefore faecal samples were collected during the last 4 months of pregnancy of Thoroughbred (n = 10), New Forest pony (n = 9), Shetland pony (n = 10) and Iceland pony mares (n = 11). Concentrations of oestrogens, 20alpha-hydroxy- and 20-oxopregnanes were measured using enzyme immunoassays. Breed differences concerning both levels (though significant only in case of oestrogens) and time course of measured steroids were observed. There was a highly significant time effect (p < 0.00001) and an interaction between time and breeds (p < 0.02) for all steroids measured, suggesting that the time effect differs for different breeds. Oestrogen concentrations showed a decrease towards parturition, whereas in 20alpha-hydroxy- and 20-oxopregnane levels a pronounced increase was found 2 and I months, respectively, before parturition. A breed effect was only significant (p = 0.001) when comparing oestrogen concentrations and was mainly due to Iceland ponies, which had the lowest concentrations especially during the last 2 months of pregnancy. An almost significant (p = 0.06) breed effect was found for 20-oxopregnanes. In Iceland mares an additional increase in faecal pregnane content was already observed earlier, reaching maximum levels before the 60th day ante-partum (a.p.), followed by a decrease until the 30th day a.p. The ratio of 20-oxopregnanes to oestrogens in the samples was significantly higher (p < 0.006) in Iceland ponies in comparison with any other breed throughout all months before parturition. The breed differences observed in the amounts of oestrogens and/or progestagens present during late pregnancy may demonstrate micro-evolutionary changes in the endocrine system of a species. PMID- 11885747 TI - The evolution of the endozepine-like peptide (ELP) in the mammalian testis. AB - The endozepine-like peptide (ELP) is a testis-specific isoform of the acyl-CoA binding protein (ACBP) and shares the latter's peptide motif for binding mid-long chain acyl-CoA groups. ELP is expressed both as mRNA and protein at high levels in the testes of a wide range of mammals, including rodents, carnivores and ruminants. However, the ELP gene is progressively inactivated through primate evolution, with no protein detectable in a range of primates studied, including human. In nonprimate species, ELP is expressed in very late postmeiotic germ cell stages only, such that its function in these species is probably associated with the metabolism of the mature spermatozoon. Current research is looking at both the function of the ELP protein and the haploid regulation of the gene. PMID- 11885749 TI - Inhibitory actions of indomethacin on electrical and mechanical responses produced by nerve stimulation in circular smooth muscle of the guinea-pig gastric fundus. AB - The effects of indomethacin on electrical and mechanical responses produced by transmural nerve stimulation (TNS) were investigated in isolated circular smooth muscle of the guinea-pig gastric fundus. TNS evoked a cholinergic excitatory junction potential (e.j.p.). The e.j.p.s were inhibited by 1-10 microM indomethacin, in a concentration-dependent manner, with no marked alteration of the resting membrane potential. Exogenously applied acetylcholine caused a depolarization of the membrane that was not altered by indomethacin. TNS evoked a cholinergic twitch contraction at low frequencies (0.1 Hz). A train of TNS's at high frequency (1 Hz) produced a transient contraction with a subsequent sustained relaxation. Indomethacin reduced the resting tension and inhibited these TNS-induced contractions. Application of Nomega-nitro-L-arginine (NOLA), an inhibitor of nitric oxide (NO) synthesis, increased the amplitude of twitch contractions, and altered transient contractions to tetanic contractions during TNS at a frequency of 1 Hz, also with an increased amplitude. In the presence of NOLA, indomethacin (5 microM) again reduced the resting tension and inhibited TNS induced contractions. This inhibition was greater for twitch contractions than for tetanic contractions. Nifedipine reduced the TNS-induced contractions, while addition of indomethacin further reduced the amplitude of contractions. Contractions produced by low concentrations of acetylcholine (0.1 microM) were inhibited by indomethacin, while those produced by 1 microM were not. These results indicate that the inhibitory actions of indomethacin on TNS-induced contractions do not involve enhanced production of NO or selective inhibition of voltage-gated Ca-channels. Prejunctional autoregulatory mechanisms may also not be altered by indomethacin. As indomethacin inhibits the enzyme cyclooxygenase, it is speculated that endogenously produced prostaglandins exert excitatory actions on gastric smooth muscle, and act mainly postjunctionally to facilitate spontaneous and neurogenic electrical and mechanical activity. PMID- 11885750 TI - A light microscope study of the distribution of muscle in the frog esophagus and stomach. AB - The present study reports light microscopical observations of the distribution of muscle in the esophagus and stomach of both the bull frog (Rana catesbeiana) and the African clawed frog (Xenopus laevis). The external muscle coat of the upper half of the esophagus in both species had several collagen coated bundles of striated muscle fibres around the circumference. These striated muscle bundles ran longitudinally from the pharynx to around the vicinity of the center of the esophagus. Beneath these striated muscle bundles was an inner circular layer of smooth muscle. In both species, the inner circular layer of smooth muscle was particularly thick in the region close to the pharynx. In the bull frog, the lower half of the esophagus lacked striated muscle. However, the circular smooth muscle layer, extending from the upper half of the esophagus, was also observed throughout the lower half of the esophagus. An outer longitudinal layer of smooth muscle developed towards the terminal portion of the esophagus such that in this region, both outer longitudinal and inner circular layers of smooth muscle were observed. Similarly in the African clawed frog, the inner circular layer of smooth muscle was continuous along the full length of the esophagus. Again, no striated muscle bundles were observed in the lower half of the esophagus. However, the outer longitudinal layer of smooth muscle was seen to develop in the middle region of the esophagus. Its muscle layer extended to the terminal portion of the esophagus. Thus, both outer longitudinal and inner circular layers of smooth muscle were observed throughout the lower half of the esophagus. In both frogs, the thickness of the outer longitudinal and inner circular layers of smooth muscle changed before and after the esophago-gastric junction. In both frogs, no muscularis mucosa was observed in the esophageal wall. However, in the lower half of the esophagus of the African clawed frog, small bundles of smooth muscle were observed here and there in the submucosa. A fully developed muscularis mucosa with both outer longitudinal and inner circular layers was observed in the upper stomach of both frogs. PMID- 11885748 TI - Structure-activity relationship studies of (+/-)-terbutaline and (+/-)-fenoterol on beta3-adrenoceptors in the guinea pig gastric fundus. AB - (+/-)-Terbutaline and (+/-)-fenoterol are both arylethanolamine analogs that have tertbutyl and aryliso-propyl substituents respectively at the a position on the nitrogen of the ethanolamine side chain. In the present study, we have investigated the structure-activity relationships of (+/-)-terbutaline and (+/-) fenoterol as beta3-adrenoceptor agonists in the guinea pig gastric fundus. (+/-) Terbutaline and (+/-)-fenoterol induced concentration-dependent relaxation of the precontracted gastric fundus with pD2 values of 4.45+/-0.10 and 5.90+/-0.09, and intrinsic activities of 1.00+/-0.03 and 0.99+/-0.01 respectively. The combination of the selective beta1-adrenoceptor antagonist (+/-)-atenolol (100 microM), and the selective beta2-adrenoceptor antagonist (+/-)-butoxamine (100 microM), produced a 2 and 6 fold rightward shift of the concentration-response curves for (+/-)-terbutaline and (+/-)-fenoterol respectively, without depressing the maximal responses. The order of potency of these agonists was (pD2 value): (+/-) fenoterol (5.09+/-0.10) > (+/-)-terbutaline (4.13+/-0.08). In the presence of (+/ )-atenolol and (+/-)-butoxamine, however, the non-selective beta1, beta2- and beta3-adrenoceptor antagonist (+/-)-bupranolol caused a concentration-dependent rightward shift of the concentration-response curves for (+/-)-terbutaline and (+/-)-fenoterol. Schild plot analyses of the effects of (+/-)-bupranolol against these agonists gave pA2 values of 6.21+/-0.07 ((+/-)-terbutaline) and 6.37+/-0.06 ((+/-)-fenoterol) respectively, and the slopes of the Schild plot were not significantly different from unity (p>0.05). These results suggest that the relaxant responses to (+/-)-terbutaline and (+/-)-fenoterol are mainly mediated through beta3-adrenoceptors in the guinea pig gastric fundus. The beta3 adrenoceptor agonist potencies of arylethanolamine analogs depend on the size of the end of the alkylamine side chain. PMID- 11885751 TI - Neonatal hydronephrosis detected on routine health check-up. AB - BACKGROUND/PURPOSE: Prenatal dilatation of the urinary tract may be caused by obstructive defects, and it is known that 20% of normal fetuses have some degree of renal pelvic or calyceal dilatation, determined by sonographic examination. We analyzed the clinical course of patients found to have dilatation of the renal pelvis by ultrasound 1 month after birth, and compared prenatal and postnatal sonograms of each patient. METHODS: Between January 1996 and December 1998, renal ultrasounds were performed on 2,071 children at their 1 month routine health checks in our hospital. We found dilatation of the renal pelvis in 92 kidneys in 84 children and then compared these neonatal sonograms with the prenatal ultrasounds for each child. Ultrasound examinations were performed at 28 weeks of gestational age and 1 month after birth. Our criterion for diagnosis of hydronephrosis at 1 month of age was a renal pelvis measuring greater than 7 mm at the central echo complex. The medical records of patients found to have hydronephrosis were then reviewed. RESULTS: Eight patients had bilateral hydronephrosis, 2 had dilatation only in the right kidney and 74 had this finding only in the left kidney. Examination of 36 (39%) of these 92 kidneys revealed renal pelvic dilatation to be present both prenatally and neonatally. The dilatation ranged from 7 to 43 mm. No dilatation of the renal pelvis was seen on the other 56 fetal examinations. During the follow-up period, 2 patients (1 with bilateral and the other with right-sided hydronephrosis) were diagnosed with vesicoureteral reflux, the 1 patient with bilateral pelvic dilatation was found to have a vesico-ureteral junction obstruction. After full evaluation, the other children were found to have no anatomic abnormalities. CONCLUSIONS: We found 84 of 2,071 children showed dilatation of the renal pelvis on ultrasound examination performed at 1 month of age. Three (3.3%) of the 84 children required surgery to correct the neonatal hydronephrosis detected via this imaging modality. Interestingly, 88% of the children had only left-sided hydronephrosis, which did not predict an adverse outcome during the follow-up period. We conclude that neonatal hydronephrosis appears to be a relatively benign condition and the requirement of surgery is relatively slight. PMID- 11885753 TI - Valsalva maneuver prevents guide wire trouble associated with 22-gauge safe guide. AB - The Safe guide is a central venous puncture needle that serves as both a pilot needle and as an introducer. A guide wire can be inserted into a vein through the side port at the hub of the 22-gauge Safe guides needle initially inserted as a pilot needle. However, guide wire insertion may fail due to kinking or locking at the side port. Increasing airway pressure to 20 cm H2O by squeezing a respiratory bag during insertion of the guide wire together with venous puncture was attempted to determine if would decrease guide wire trouble. SUBJECTS AND METHODS: A total of 120 patients scheduled for central venous catheterization by right internal jugular puncture were divided into two groups. Patients in group-A (n = 60) were catheterized by the conventional method and those in group-B (n = 60) were catheterized by applying the Valsalva maneuver. Three observations were made: 1) Frequency of cases in which blood back-flow occurred during withdrawal only and not upon advancement of the puncture needle. 2) Frequency of cases in which kinking and/or locking of the guide wire occurred at the hub during its insertion. And 3) the occurrence of complications. RESULTS: 1) The patency of the vein was preserved and blood back-flow was obtained during advancement of the puncture needle in all cases in which the Valsalva maneuver was applied. 2) The incidence of kinking and/or locking during insertion of the guide wire decreased from 16.7% to 3.4% by applying positive airway pressure during the Valsalva maneuver. And 3) complications were negligible. Additionally, the application of the Valsalva maneuver allowed successful guide wire insertion in 6 out of 9 cases (67%) in group-A, in which the initial attempt using the conventional method had failed. CONCLUSION: The application of positive airway pressure using the Valsalva maneuver may prevent the guide wire trouble associated with the 22-gauge Safe guide. PMID- 11885752 TI - Influence of body fat on the onset of vecuronium induced neuromuscular blockade. AB - The onset time of vecuronium, a muscle relaxant, was measured after a bolus intravenous injection of 0.15 mg kg(-1) of vecuronium into 40 surgical patients aged 59-64 years. The onset time was then compared between male and female patients and the relationship between onset time and body fat (% of body weight) was analyzed. Arterial plasma concentrations of vecuronium were measured at 75, 195, and 375 sec after administration of vecuronium to 8 patients. The female patients (n = 23) showed a shorter onset time and more body fat than the male patients (n = 17). The onset time significantly decreased with increasing body fat in both groups. When only females with body fat of less than 30% (n = 10) were compared with the male group (all male patients had body fat of less than 30%), the body fat, onset time, and regression lines between the onset time and the body fat did not differ significantly. Except in the patient with the highest body fat, plasma concentrations at 195 and 375 sec significantly increased with increasing body fat. We concluded that the higher body fat in females is largely responsible for the faster onset of vecuronium action in females. A smaller distribution volume of vecuronium may also be one of the reasons for the faster onset of vecuronium in females. PMID- 11885754 TI - The E4 allele of apolipoprotein E is associated with increased restenosis after coronary angioplasty. AB - The aim of this study is to investigate the influence of the E4 allele of apolipoprotein E (apo E) on restenosis after percutaneous transluminal coronary angioplasty (PTCA). The subjects were 171 male patients with more than 75% luminal diameter stenotic lesions of the coronary artery who had undergone an elective initial PTCA. The PTCA was successful in 164 patients, 157 of whom completed a prospective 5 month coronary angiography (CAG) follow up to assess the degree of restenosis after their surgery. Patients with previous coronary artery bypass grafting surgery (CABG), 3 vessel disease, complete obstruction or calcified lesions of the coronary artery, cerebro-vascular disease (CVD), arteriosclerosis obliterans (ASO), and renal failure with hemodialysis were excluded, leaving 105 patients in the analysis. Subjects carrying the E4 allele (n = 22, Phenotype E4/2 = 2, E4/3 = 19, E4/4 = 1: E4 group) were well matched with non-carriers (n = 83, Phenotype E2/2 = 0, E3/2 = 4, E3/3 = 79: E3 group) for clinical, and pre-and post-PTCA angiographic features. The restenosis rates were significantly higher in the E4 group than in the E3 group (patient restenosis rate : 59.1 vs 33.7% p < 0.05, lesion restenosis rate: 51.8 vs 30.9% p < 0.05). These results suggest that the E4 allele is associated with a higher restenosis rate after PTCA. PMID- 11885755 TI - Effect of diabetic retinopathy on redox state of aqueous humor and serum albumin in patients with senile cataract. AB - PURPOSE: To investigate the oxidative status of albumin in the aqueous humor and serum of senile cataract patients with diabetes in order to clarify the pathogenesis of this condition. METHODS: High-performance liquid chromatography (HPLC) was employed to measure the reduced form of albumin (mercaptalbumin) and the oxidized form of albumin (nonmercaptalbumin) in serum and aqueous humor. The mercaptalbumin, nonmercaptalbumin-1, and nonmercaptalbumin-2 fractions in aqueous humor obtained at the start of cataract surgery and in serum obtained intraoperatively were analyzed by HPLC in 7 senile cataract patients with diabetic retinopathy (2 men and 5 women aged 70+/-9.8 years). RESULTS: The mean content (%) of mercaptalbumin, nonmercaptalbumin-1, and nonmercaptalbumin-2 in serum albumin from the diabetic patients was 60.3+/-7.8, 36.9+/-6.6, and 2.7+/ 1.7%, respectively, while the corresponding values for aqueous humor albumin were 40.0+/-14.1, 52.4+/-12.6, and 7.5+/-3.6%. When the mercaptalbumin content (%) of aqueous humor albumin was compared between patients with active and inactive diabetic retinopathy, the respective values were 47.0+/-9.1% and 22.8+/-5.7%. A significant correlation mercaptalbumin content (%) of aqueous humor albumin did not show with the HbA1c level, but there was still a relationship (Y = 5.0 x - 2.7, r = 0.80, and p < 0.052). CONCLUSION: The increase of mercaptalbumin (the reduced form of albumin) in the aqueous humor of patients with diabetic retinopathy probably resulted from an increase of retinal vascular permeability. PMID- 11885756 TI - Structural equation modeling of determinants of planning. AB - A model is proposed that implies that planning evoked by the formation of an implementation intention is related to behavioral intention and perceived behavioral control, whereas in accordance with the theory of planned behavior, behavioral intention is related to attitude and perceived behavioral control. Measures of attitude toward the behavior, perceived behavioral control, behavioral intention, and planning were constructed from 192 undergraduates' ratings of descriptions of two fictitious situations in which a target behavior was varied with respect to benefit and actual behavioral control. Structural equation modeling yielded an acceptable fit of the proposed model. PMID- 11885757 TI - The measurement of core affect: a Swedish self-report measure derived from the affect circumplex. AB - Three studies were conducted with the aim of developing a new Swedish self-report measure of core affect (the Swedish Core Affect Scale or SCAS). In Study 1,122 participants rated their current mood on 24 unipolar adjective scales. A revised set of 12 bipolar adjective scales was evaluated in Study 2 employing 96 participants who rated their current mood before and after a mood-inducing naturally occurring event. A slightly revised set of adjective scales was used in Study 3, in which another 96 participants rated several induced moods. The results showed that the adjective scale ratings could be aggregated as reliable measures of the independent valence and activation dimensions proposed in the affect circumplex, and that the aggregated measures discriminated mood differences within and between individuals. PMID- 11885758 TI - The auditory sensory memory trace decays rapidly in newborns. AB - The present study investigated the temporal dynamics of auditory sensory memory in newborns as reflected by the mismatch negativity (MMN), a preattentive electric change-detection response. MMN was obtained from 24 full-term healthy newborns who were either awake or asleep (quiet or active sleep) during the experiments. Stimuli were 1,000 Hz tones (standards) that were occasionally replaced by 1,100 Hz tones (deviants). The constant stimulus onset asynchrony (SOA) was, in separate blocks, either 450, 800, or 1,500 ms. A prominent MMN was obtained at the 800 ms SOA in all three sleep or waking states, whereas no MMN occurred at 450 and 1,500 ms SOAs. In view of the fact that in adults MMN is elicited even with a 10s SOA, these results imply that the time span of auditory memory is considerably shorter in neonates than in adults and 8-12-year-old children. PMID- 11885759 TI - Prenatal drug exposure and the conceptualization of long-term effects. AB - This paper discusses several factors affecting the development of children prenatally exposed to drugs. In the "first generation" of research in this field a main factor model of disease formed the basis for a belief in the feasibility of detecting the direct pharmacological or teratogenic effects of drug exposure on long-term child development. However, the clustering of confounding variables has constituted a major problem in identifying these effects. In the last few years a "second generation" of research in this field has emerged, and investigators have moved beyond simple main-effect models. The importance of controlling for confounding variables has been underscored. However, prenatal substance exposure is still often studied within a teratology model where the main goal is the search for unique effects of a specific drug or substance. Based on this review it is suggested that an appropriate model for understanding the development of drug-exposed children cannot be based on a main-effect perspective. Rather, such a model must evolve from a contextual perspective, and it is suggested that a transactional model, where both potential risk factors and protective factors are considered, should replace the traditional teratology model in this field. PMID- 11885760 TI - The hierarchical structure of empathy: dimensional organization and relations to social functioning. AB - The purpose of the present study was to examine the structure of empathy using a hierarchical approach, and to compare the dimensions of empathy with measures of social functioning, in order to contribute to the understanding of the nature of empathy. The dimensionality of the Interpersonal Reactivity Index, which comprises four subscales (empathic concern, perspective taking, fantasy and personal distress) was examined using confirmatory factor analysis. Relations with the Social Skills Inventory were also investigated. A sample of 127 applicants for places on nursing and social work undergraduate programs participated in the study. The study findings indicate that empathy is hierarchically organized, with one general dimension at the apex. The general factor is identical to empathic concern and this dimension overlaps to a great extent with perspective taking and fantasy. The findings also indicate that the general dimension constitutes an integrated entirety, with its main emphasis on emotional reactivity by also involving cognitive processes. PMID- 11885761 TI - Temperament in children with Down syndrome and in prematurely born children. AB - Parents of three groups of children completed the Children's Behavior Questionnaire (CBQ). Participants were children with Down syndrome aged 4-11 years (n = 55), prematurely born children aged 5 years (n = 97), and a group of normally developing kindergarten children 5-7 years of age (n = 91). Mean levels and factor structures on the CBQ were compared between the three groups. The children with Down syndrome had less attentional focusing and expressed less inhibitory control and less sadness than the normally developing children. There were also group differences in temperament structures, especially a clearer emotional factor of "surgency" among the children with Down syndrome. The only significant difference in mean temperament scores between the premature children and the control group was that the former evinced less attentional focussing. The temperament structures in the Norwegian samples were very similar to those reported in earlier studies, conducted in China and the US. PMID- 11885762 TI - Tactile memory of deaf-blind adults on four tasks. AB - The performance of ten deaf-blind and ten sighted-hearing participants on four tactile memory tasks was investigated. Recognition and recall memory tasks and a matching pairs game were used. It was hypothesized that deaf-blind participants would be superior on each task. Performance was measured in terms of the time taken, and the number of items correctly recalled. In Experiments 1 and 2, which measured recognition memory in terms of the time taken to remember target items, the hypothesis was supported, but not by the length of time taken to recognize the target items, or for the number of target items correctly identified. The hypothesis was supported by Experiment 3, which measured recall memory, with regard to time taken to complete some of the tasks but not for the number of correctly recalled positions. Experiment 4, which used the matching pairs game, supported the hypothesis in terms of both time taken and the number of moves required. It is concluded that the deaf-blind people's tactile encoding is more efficient than that of sighted-hearing people, and that it is probable that their storage and retrieval are normal. PMID- 11885763 TI - Structure of conduct problems in adolescence. AB - Scholars disagree about whether adolescent conduct problems (CPs) form a single behavioral syndrome or whether such problems are better conceptualized as different dimensions. The arguments raised by both sides are addressed and tested empirically by analysing data from a large general population sample of Norwegian adolescents (n = 9,342). Confirmatory factor analyses show that a single syndrome of CPs may be subdivided into three highly correlated factors. The first dimension, destructive covert, includes theft and vandalism, whereas the second, nondestructive covert, reflects avoidance of arenas under adult control. The last dimension, overt, includes school opposition and fighting. This three-factor model fits well for both girls and boys, and individuals in their early and late teens. Results from scale analyses are modestly to moderately indicative of a developmental continuum of severity of CPs ranging from nondestructive covert to overt to destructive covert. Taken together the findings imply that both a unidimensional and a multidimensional perspective on CPs are applicable in the general youth population. PMID- 11885764 TI - Cultural changes (1986-96) in a Norwegian airline company. AB - The purpose of the study was to investigate cultural changes in a Norwegian airline company over a time span of 10 years. A questionnaire including parameters characterizing culture was administered to air crews in 1986 (n = 137) and in 1996 (n = 50). The performance part of a simulator study in 1996 indicated a significant reduction in operational failures compared with the 1986 study. The data further demonstrated significant changes in cultural variables, such as reduced Dominance and Masculinity, and improved Social climate and Communication. The direction of change in scores on the cultural variables corresponded with the principles on which the remedial actions were based. PMID- 11885765 TI - The pig as a source of cardiac xenografts. AB - The inadequate availability of human donor hearts and other organs has inspired interest in the field of xenotransplantation. Historically, ten attempts to transplant animal hearts into human recipients have been reported. Those who received hearts from nonhuman primates (i.e., baboons and chimpanzees) survived rather longer than did those who received hearts from nonprimates (i.e., sheep and pigs). Nevertheless, current opinion is that the pig is the best candidate as a source of hearts for humans despite the considerable immunologic disparity between the two species. Pigs are available in large numbers and can be bred easily and rapidly. They grow to appropriate sizes and their cardiovascular system is similar to that of humans. Substantial knowledge has been accumulated regarding both genetic engineering and tolerance induction in pigs, two strategies that may help to overcome the existing immunologic barriers. Concern has been raised, however, with regard to the potential for the transfer of a porcine infection with the pig organ to the human recipient. This brief review addresses these and other aspects of the use of the pig as a source of hearts for patients with end-stage cardiac disease. PMID- 11885766 TI - The pathology of cardiac xenografts. AB - The pathology of cardiac xenografts has yielded critical insights into the mechanisms of xenograft rejection and the therapeutic procedures that might be applied to preventing or treating it. The conditions seen in rejecting cardiac xenografts include hyperacute rejection, acute vascular rejection, and cellular rejection. Hyperacute and acute vascular rejection of cardiac xenografts have features typical of humoral injury. Less is known about cellular rejection and only speculation can be offered about chronic rejection. Still, these features allow critical testing of pathogenetic mechanisms and therapies. PMID- 11885767 TI - Infection in xenotransplantation. AB - Advances in transplantation immunology have enhanced the possibility of xenotransplantation as a therapeutic option for end-stage organ failure. The potential spread of animal-derived pathogens to the recipient and to the general population, termed "xenosis," is a potential complication of interspecies transplantation. Recognition of such novel infections may be complicated by infections due to altered microbiologic behavior and clinical symptomatology of these organisms, particularly in the immunocompromised xenograft recipient. Particular concern exists over the activation of latent viruses, including retroviruses, from xenograft tissues. Based on experience with human allogeneic transplantation, those pathogens considered most likely to cause human disease can be excluded prospectively from herds of animals developed for organ donation. Research is needed into the activation and behavior of retroviruses and other potential pathogens in xenotransplantation. Xenotransplantation may also provide unique opportunities not only for the care of patients with organ failure, but in the therapy of individuals with chronic infections to which the xenograft may be resistant. Clinical protocols must be developed so as to enhance the safety of the recipient and of the community-at-large. PMID- 11885768 TI - Therapeutic strategies for xenograft rejection. AB - The increasing demand for transplantable organs over the past several decades has stimulated the idea of using animal organs in lieu of cadaveric organs in clinical transplantation. Pigs are now considered to be the most suitable source of organs for transplantation because of their abundant availability, their appropriate size, their relatively short gestation period, and the recent development in the technology to genetically manipulate them. In the past few years, some of the seemingly complex immunologic responses in pig-to-primate transplantation have been elucidated. This progress has allowed us to focus our efforts on devising specific therapeutic strategies to overcome or prevent some of the responses that contribute to rejection of the xenograft. In this article, we review the various approaches that might allow clinical xenotransplantation to come to fruition. PMID- 11885769 TI - Concordant cardiac xenotransplantation. PMID- 11885770 TI - Surgical treatment of acute infective valvular endocarditis (18 years experience). AB - BACKGROUND: Infective endocarditis morbidity remains high: 3 to 8 cases per 100,000 of population. Antibiotic therapy is ineffective. Its surgical treatment experience is relatively limited. AIM: To share the surgical treatment experience of 855 patients with acute infective valvular endocarditis (AIVE) treated during 1982 to 2000 in the Institute of Cardiovascular Surgery AMS, Ukraine. MATERIALS AND METHODS: 855 (75.4%) of 1128 hospitalized patients with AIVE were operated upon. Surgical interventions included removal of diseased tissues, heart chambers treatment with antiseptic solutions, wash out with normal saline solution, replacement or plastic procedure of valves. RESULTS: Heart abscesses were found in 132 (15.5%) patients. Hospital mortality was after aortic valve replacement 12.6%; mitral valve replacement 9.7%; plastic procedure on mitral valve 0%; aortic and mitral valve replacement 30%; tricuspid valve replacement 15.4%; and plastic procedure on tricuspid valve 6.1%. Recurrences of infective process occurred in 51 (6.0%) patients. Infections were observed more frequently in patients with heart abscesses: 10.6% versus 5.7% (p < 0.02). RESULTS: 716 (96.7%) patients were studied 2 to 194 (87.4+/-39.4) months postoperatively. Tenth year postoperative survival was 62.1+/-27.7% including hospital mortality. CONCLUSIONS: (1) AIVE has become one of the most frequent causes of acquired heart lesions in the postChernobyl nuclear power station catastrophe era. (2) Heart failure development in postoperative period is stipulated by the disease duration. (3) Presence of heart abscesses favors recurrence of development of infective endocarditis. (4) Postoperative antibiotic therapy for more than 3 weeks does not help in prevention of recurrences. PMID- 11885771 TI - Evaluation of kangaroo aortic valved conduits in a juvenile sheep model: preliminary findings. AB - BACKGROUND: Biological heart valve substitutes, manufactured from either porcine or bovine tissue, have been in use for more than 30 years. Despite low thrombogenicity and excellent performance, bioprosthetic heart valves tend to degenerate and calcify early in young patients because of patient and valve related factors. The aim of this study was to examine the calcification behavior of glutaraldehyde-preserved kangaroo heart valves in a juvenile sheep model. METHODS: Porcine (n = 10) and kangaroo (n = 10) valved conduits were implanted in the descending aortic position of juvenile sheep and retrieved after 6, 8, and 12 months. Retrieved valved conduits were examined for morphological changes and calcification of the valve tissue, using Von Kossa's stain technique and atomic absorption spectrophotometry. RESULTS: Structural valve deterioration, characterized by increased stiffness and severe calcification, occurred in 100% of the porcine conduits within 4 months. Kangaroo valve leaflets were significantly (p < 0.001) less calcified at 6 months (3.39+/-1.80 microg/mg), 8 months (5.86+/-4.57 microg/mg), and at 12 months (14.38+/-6.72 microg/mg), compared to porcine valves at 3 months (176.45+/-42.88 microg/mg ) and at 4 months (154.67+/-52.67 microg/mg ). Porcine aortic wall tissue was more calcified (118.24+/-42.86 microg/mg) than kangaroo aortic wall tissue (79.55+/-26.40 microg/mg). CONCLUSIONS: Kangaroo heart valves calcify less than porcine heart valves. These findings suggest that a different donor valve tissue has a lower calcification potential probably due to a difference in the morphological ultrastructure. This could result in improved long-term durability of kangaroo heart valves. PMID- 11885772 TI - Open heart surgery in patients with dialysis-dependent renal insufficiency. AB - OBJECTIVE: Chronic renal failure (CRF) is commonly considered a significant factor for increased morbidity and mortality after cardiac surgery. METHODS: To assess the risk in our population we retrospectively analyzed 28 patients (16 men and 12 women, mean age 58.1+/-10.8 years) with end-stage renal disease (ESRD) undergoing cardiac surgery between 1989 and 2001. Sixteen (57.2%) patients had isolated coronary artery bypass grafting (CABG), nine (32.1%) had isolated valve replacement, and three (10.7%) underwent combined CABG and valve replacement. Preoperatively, 20 (71.4%) patients were on hemodialysis and eight (28.6%) on peritoneal dialysis. Mean preoperative duration of dialysis was 38.7+/-24.9 months (range, 3 to 93 months). RESULTS: There were two perioperative deaths (30 day mortality, 7.1%). Actuarial survival at 1, 2, 5, and 12 years was 0.85+/-0.7, 0.73+/-0.10, 0.65+/-0.12, and 0.54+/-0.14, respectively. Among 22 survivors, mean NYHA class was 1.7+/-0.8 (p < 0.001 vs. preoperatively) and mean CCS class was 1.6+/-0.6 (p < 0.001 vs. preoperatively). CCS/NYHA functional class IV (p = 0.01), urgent/emergency operation (p < 0.001), LVEF < 35% (p < 0.001) were strongly related to early and late mortality. CONCLUSIONS: Open-heart operations can be performed with acceptable short- and long-term results in patients with CRF on dialysis. Adequate preoperative management with identification of high risk patients and a more aggressive approach before the onset of symptoms of cardiac failure are advisable. PMID- 11885773 TI - Techniques for lengthening vein grafts in coronary artery bypass grafting. PMID- 11885774 TI - Disseminated cholesterol embolism after coronary artery bypass grafting. AB - Blue toe syndrome caused by cholesterol emboli is a relatively benign disease. However, disseminated cholesterol embolism is a life-threatening condition. We describe here the case of a 71-year-old female admitted because of anterior chest pain and intermittent claudication. Following cardiac catheterization, warfarin potassium was administered. However, the patient's toes soon darkened bilaterally, and BUN and creatinine levels increased from the normal value. Skin discoloration and renal failure were improved after stopping warfarin potassium administration. The patient underwent coronary artery bypass grafting and left femoropopliteal bypass. Cerebral infarction and renal failure occurred postoperatively due to disseminated cholesterol embolism. The patient died from renal failure on the 16th postoperative day without regaining consciousness following surgery. For high risk patients, interventional procedures to the ascending aorta must be avoided. When CABG cannot be avoided for coronary revascularization, off-pump bypass and use of arterial grafts are recommended. PMID- 11885775 TI - Current status of arterial revascularization. PMID- 11885776 TI - Quantification of free fatty acids in human cerebrospinal fluid. AB - Free fatty acids (FFA) in cerebrospinal fluid (CSF) are well-recognized markers of brain damage in animal studies. Information is limited regarding human CSF in both normal and pathological conditions. Samples of CSF from 73 patients, who had undergone lumbar puncture for medically indicated reasons, came from a core laboratory upon completion of ordered tests. Using high performance liquid chromatography, mean FFA concentrations (microg/L +/- SEM) were: arachidonic 26.14 +/- 3.44; docosahexaenoic 60.74 +/- 5.70; linoleic 105.07 +/- 10.98; myristic 160.38 +/- 16.17; oleic 127.91 +/- 10.13; and palmitic 638.34 +/- 37.27. No differences in FFA concentrations were seen with gender, race, age, and/or indication for lumbar puncture. This is the first study to document normal human CSF FFA concentrations in a large series. Further characterization of FFA in pathological conditions may provide markers for evaluating clinical treatments and assisting in prognostication of neurological disease. PMID- 11885777 TI - Presynaptic modulation of K+-evoked [3H]dopamine release in striatal and frontal cortical synaptosomes of normotensive and spontaneous-hypertensive rats. AB - Regional differences in presynaptic [3H]dopamine ([3H]DA) release and its modulation by D2 DA-receptors between the frontal cortex and striatum obtained from Wystar-Kyoto (WKY) and spontaneous-hypertensive rats (SHR) have been evaluated using superfused synaptosomes. Synaptosomal tritium content was significantly lower in the frontal cortex than in the striatum in both SHR and WKY (approximately 45% and 48%, respectively), but no differences in tritium content were obtained between strains. However, the 15 mM K+-evoked [3H]DA overflow was lower in the SHR as compared to WKY rats in both brain regions (striatum approximately 23%, frontal cortex approximately 21). Concentration response curves for quinpirole (1nM-10 microM)-mediated inhibition of 15mM K+ evoked [3H]DA release showed no differences between SHR and WKY. These results suggest that SHR has less ability to release [3H]DA as compared to WKY rats, but SHR did not show differences in the autoregulation of such release in both the frontal cortex and striatum. PMID- 11885778 TI - L-pyroglutamic acid inhibits energy production and lipid synthesis in cerebral cortex of young rats in vitro. AB - In the present study we investigated the effects of L-pyroglutamic acid (PGA), which predominantly accumulates in the inherited metabolic diseases glutathione synthetase deficiency (GSD) and gamma-glutamylcysteine synthetase deficiency (GCSD), on some in vitro parameters of energy metabolism and lipid biosynthesis. We evaluated the rates of CO2 production and lipid synthesis from [U-14C]acetate, as well as ATP levels and the activities of creatine kinase and of the respiratory chain complexes I-IV in cerebral cortex of young rats in the presence of PGA at final concentrations ranging from 0.5 to 3 mM. PGA significantly reduced brain CO2 production by 50% at the concentrations of 0.5 to 3 mM, lipid biosynthesis by 20% at concentrations of 0.5 to 3 mM and ATP levels by 52% at the concentration of 3 mM. Regarding the enzyme activities, PGA significantly decreased NADH:cytochrome c oxireductase (complex I plus CoQ plus complex III) by 40% at concentrations of 0.5-3.0 mM and cytochrome c oxidase activity by 22-30% at the concentration of 3.0 mM, without affecting the activities of succinate dehydrogenase, succinate:DCPIP oxireductase (complex II), succinate:cytochrome c oxireductase (complex II plus CoQ plus complex III) or creatine kinase. The results strongly indicate that PGA impairs brain energy production. If these effects also occur in humans, it is possible that they may contribute to the neuropathology of patients affected by these diseases. PMID- 11885779 TI - Influence of convulsants on rat brain activities of alanine aminotransferase and aspartate aminotransferase. AB - There exist differences between 12-day-old and adult rats in the onset of seizures induced by some inhibitors of glutamate decarboxylase (GAD). The aim of study was to investigate if there are differences between both groups in activities of rat brain alanine aminotransferase (ALT) and aspartate aminotransferase (AST), the enzymes involved in glutamate metabolism, after the administration of 3-mercaptopropionic acid as specific GAD inhibitor or isoniazid as less specific general inhibitor of pyridoxal enzymes. Activities of both aminotransferases in a supernatant 20,000 g of the whole brain (containing predominantly cytosolic isoforms of enzymes) were increased at the beginning of 3 mercaptopropionic acid-induced generalized tonic-clonic seizures. At isoniazid induced generalized tonic-clonic seizures, a significant increase in both enzyme activities was observed in adult rat brain. In the 12-day-old rat brain, ALT and AST activities reached about 40% and about 50-60% of adult control levels, respectively. In in vitro experiments, no influence of 3-mercaptopropionic acid on transaminase activities was found and an inhibitory effect of isoniazid on the enzymes was confirmed. Increased aminotransferase activities might participate in the enhanced synthesis of excitatory amino acid neurotransmitters in the nervous system, which may take a part in the initiation of epileptic seizures. Alternatively, the increased AST activity may be connected with an increased transport of NADH from the cytosol to mitochondria, while the increased ALT activity would represent the transformation of pyruvate to alanine as a consequence of increased glycolysis. PMID- 11885780 TI - Effects of glia maturation factor overexpression in primary astrocytes on MAP kinase activation, transcription factor activation, and neurotrophin secretion. AB - Using the replication-defective adenovirus vector, we overexpressed rat glia maturation factor (GMF) in primary astrocyte cultures derived from embryonic rat brains. Among the three isoforms of MAP kinase, there was a big increase in the phosphorylation of p38, as detected with Western blotting using the phosphospecific antibody. Likewise, there was a substantial increase in the phosphorylation of the transcription factor CREB. Using the electrophoretic mobility shift assay (EMSA), we found a stimulation in the transcription factor NF-kappaB. The activations of CREB and NF-kappaB were blocked by inhibitors of either p38 (SB-203580) or MEK (PD-098059), suggesting that they were events downstream of MAK kinase. There was an increased secretion of BDNF and NGF into the conditioned medium, along with an increase in their messenger RNA. The inductions of BDNF and NGF were also blocked by inhibitors of p38 and MEK, as well as by the inhibition of NF-kappaB with a decoy DNA sequence. Taken together, the results suggest that GMF functions intracellularly in astrocytes as a modulator of MAP kinase signal transduction, leading to a series of downstream events including CREB and NF-kappaB activation, resulting in the induction and secretion of the neurotrophins. PMID- 11885781 TI - Cell body size and succinate dehydrogenase activity of spinal motoneurons innervating the soleus muscle in mice, rats, and cats. AB - The cell body sizes and succinate dehydrogenase (SDH) activities of motoneurons in the retrodorsolateral region of the ventral horn in the spinal cord innervating the soleus muscle in mice, rats, and cats were compared using quantitative enzyme histochemistry. There was an inverse relationship between cell body size and SDH activity of motoneurons in the three species. The mean cell body sizes of both gamma and alpha motoneuron pools were in the rank order of mice < rats < cats, while the mean SDH activities of both gamma and alpha motoneuron pools were in the rank order of mice > rats > cats. It is concluded that smaller motoneurons innervating the soleus muscle have higher SDH activities than larger motoneurons, irrespective of the species, and that motoneuron pools innervating the soleus muscle in smaller animals have smaller mean cell body sizes and higher mean SDH activities than those in larger animals. PMID- 11885782 TI - Effect of dithiol chelating agents on [3H]MK-801 and [3H]glutamate binding to synaptic plasma membranes. AB - 2,3-Dimercaptopropanol (BAL- British Anti-Lewesite) is a dithiol chelating agent used for the treatment of heavy metal poisoning, however, BAL can produce neurotoxic effects in a variety of situations. Based on the low therapeutic efficiency of BAL other dithiols were developed and DMSA (meso-2,3 dimercaptosuccinic acid) and DMPS (2,3-dimercaptopropane-1-sulfonic acid) are becoming used for treatments of humans exposed to heavy metals. In the present investigation the effect of dithiols in the glutamatergic system was examined. The results showed that BAL inhibited [3H]MK-801 and [3H]glutamate binding in a concentration-dependent manner. At 100 microM BAL and DMSA caused a significantly inhibition of [3H]MK-801 binding to brain membranes (p < 0.05 by Duncan's multiple range test). BAL at 100 microM caused an inhibition of 40% on [3H]glutamate binding. DMPS and DMSA had no significant effect on [3H]glutamate binding. Dithiotreitol (DTT), abolished the inhibitory effect of BAL on [3H]MK 801 binding. The protection exerted by DTT suggests that BAL inhibit [3H]MK-801 binding by interacting with cysteinyl residues that are important for redox modulation of receptor responses. ZnCl2 inhibited [3H]glutamate and [3H]MK-801 binding to brain synaptic membrane; nevertheless, the inhibitory effect was slight more accentuated for [3H]MK-801 than [3H]glutamate binding (p < 0.05). The inhibition caused by 10 microM ZnCl2 on [3H]MK-801 binding was attenuated by BAL. The findings present in this study may provide the evidence that BAL affect the glutamatergic system and these effects can contributed to explain, at least in part, why BAL, in contrast to DMPS and DMSA is neurotoxic. PMID- 11885783 TI - Opposite effects of lithium on proximal and distal caspases of immature and mature primary neurons correlate with earlier paradoxical actions on viability. AB - To provide an explanation for earlier paradoxical findings of lithium on survival of mature and immature neurons, this study monitors changes in cytosolic caspases in rat cerebellar granule cells (CGC) grown 2-7 days in vitro (DIV), or in murine E-17 cortical neurons. Data show Li+ protects mature 7-DIV CGC parallel to a decrease in proximal and distal caspases but increases levels for immature 2-DIV CGC or E-17 cortical neurons. Caspases mirror viability based on morphological analyses (dye uptake, phase-contrast, DNA fragmentation), and suggest protection occurs by suppressing activation of a cascade resulting in distal effectors that destroy proteins essential for neuronal survival. Protection was dose-dependent with EC50 3.0 mM and extended to 64 h in K+-serum deprived apoptotic media. Neuronal extracts contain a spectrum of proximal (-2, -8, -9) and distal (-3, -6) caspases sensitive to Li+ on assay with preferred peptide substrates and by immunoblotting. The lack of direct effect on activated cytosols indicates Li+ acts upstream only on intact cells, at sites for recruitment of pivotal procaspases. Alterations of procaspase-9 p46 and membrane-bound cytochrome c (Apaf-1) point to interaction with an intrinsic Mt-mediated pathway as one of the targets. The opposite effects on caspases and viability of immature or embryological neurons point to existence of alternative pathways that alter during neurite outgrowth suggesting the use of Li+ as a probe to unravel events relevant to neurogenesis. PMID- 11885784 TI - Inhibition of Na+,K+-ATPase activity from rat hippocampus by proline. AB - Na+,K+-ATPase and Mg2+-ATPase activities were determined in the synaptic plasma membranes from hippocampus of rats subjected to chronic and acute proline administration. Na+,K+-ATPase activity was significantly reduced in chronic and acute treatment by 33% and 40%, respectively. Mg2+-ATPase activity was not altered by any treatment. In another set of experiments, synaptic plasma membranes were prepared from hippocampus and incubated with proline or glutamate at final concentrations ranging from 0.2 to 2.0 mM. Na+,K+-ATPase, but not Mg2+ ATPase was inhibited (30%) by the two amino acids. In addition, competition between proline and glutamate for the enzyme activity was observed, suggesting a common binding site for these amino acids. Considering that Na+,K+-ATPase activity is critical for normal brain function, the results of the present study showing a marked inhibition of this enzyme by proline may be associated with the neurological dysfunction found in patients affected by type II hyperprolinemia. PMID- 11885787 TI - Appropriate vision standards for safe driving: experience of on-road driving assessment. PMID- 11885785 TI - Investigation of extracellular L-citrulline concentration in the striatum during alcohol withdrawal in rats. AB - In this study, changes in striatal extracellular L-citrulline concentrations were investigated hourly for 5 h following alcohol withdrawal in chronic alcohol feeding Wistar rats. Alcohol (7.2% ethyl alcohol, v/v) was given to rats as modified liquid diet for 20 days. Signs of alcohol withdrawal appeared from the 1st h of alcohol withdrawal and the total alcohol withdrawal scores remained higher during the course of experiments. The mean of basal levels of L-citrulline in the microdialysis samples collected in conscious rat model from the striatum of control and alcoholized rats were found to be 1.28 +/- 0.48 microM and 0.35 +/ 0.08 microM, respectively. L-citrulline levels in the striatum of alcoholized rats increased by 4 folds significantly within 1 h following alcohol withdrawal. The increased striatal L-citrulline concentration was blocked by NG-nitro-L arginine methyl ester (L-NAME; 60 mg/kg), a nitric oxide synthase inhibitor, pretreatment. Our results indicate an increased L-citrulline level in the rat striatum during early alcohol withdrawal and this situation may be related to an increased nitric oxide production. PMID- 11885788 TI - Four-year review of open eye injuries at the Royal Adelaide Hospital. AB - PURPOSE: To review the epidemiology of penetrating eye injuries and ruptured globes presenting to the Royal Adelaide Hospital, South Australia. METHODS: A retrospective case review over a 4-year period. RESULTS: There were 109 penetrated or ruptured globes in 105 patients. The average age was 41 years and 80.2% were men. Over half were from rural areas. The commonest cause of injury was hammering metal followed by motor vehicle accidents. Falls in the elderly were the commenest cause of globe ruptures. A final visual acuity of 6/12 or better was found in 40% of eyes and no perception of light in 27%. CONCLUSIONS: At the Royal Adelaide Hospital, the predominant referral centre for serious ocular injury in South Australia, approximately 25 open globe injuries are encountered a year. Although the epidemiology of these injuries was found to be similar to those previously reported in Victoria and rural New South Wales, differences were thought to reflect to the ageing population of South Australia. Rupture of an old, healed large-incision cataract extraction wound was the commonest cause of ruptured globe. An effective preventive strategy to reduce the incidence of severe ocular trauma has yet to be implemented. The concept of a national population-based severe ocular trauma database is considered. PMID- 11885786 TI - Effect of graded hypoxia on high-affinity Ca2+-ATPase activity in cortical neuronal nuclei of newborn piglets. AB - Previous studies have shown that nuclear calcium signals control a variety of nuclear functions including gene transcription, DNA synthesis, DNA repair and nuclear envelope breakdown. The present study tested the hypothesis that the activity of the neuronal nuclear high affinity Ca2+-ATPase increases as a function of decreased energy metabolism in the cerebral cortex. Studies were performed in 11 ventilated newborn piglets, age 3-5 days, divided into normoxic (Nx, n = 4) and hypoxic (Hx, n = 7) groups. The animals were exposed to a single FiO2 in the range from 0.21 to 0.05 for one hr. Cerebral tissue hypoxia was confirmed biochemically by determining brain tissue ATP and phosphocreatine levels. Neuronal nuclei were isolated and the high-affinity Ca2+-ATPase activity determined. During graded hypoxia, cerebral tissue ATP decreased from 4.80 +/- 0.58 (normoxic) to 1.03 +/- 0.38 (ranging from 0.61-1.63) micromol/g brain (p < 0.05) and PCr decreased from 3.94 +/- 0.75 (normoxic) to 0.99 +/- 0.27 (ranging from 0.50 to 1.31) micromol/g brain (p < 0.05). The total high affinity Ca2+ ATPase activity in the hypoxic nuclei increased and ranged from 541 to 662 nmol/mg protein/hr, compared to activity in normoxic group of 327 to 446 nmol/mg protein/hr. During graded hypoxia, the level of nuclear high affinity Ca2+-ATPase activity correlated inversely with ATP (r = 0.91) and PCr levels (r = 0.82), with activity increasing as tissue high energy phosphates decreased. The results demonstrate that the decrease in cerebral energy metabolism during hypoxia is linearly correlated with an increase in activity of high affinity Ca2+-ATPase in cerebral cortical nuclei from immature brain. We propose that increased nuclear membrane high affinity Ca2+-ATPase activity, leading to increased nuclear Ca2+, will result in altered expression of apoptotic genes that could initiate programmed neuronal death. PMID- 11885789 TI - A comparison of cataract surgery under topical anaesthesia with and without intracameral lignocaine. AB - PURPOSE: To compare the efficacy of intracameral unpreserved lignocaine with placebo during cataract surgery under topical anaesthesia. METHODS: One hundred and thirty-five consecutive cases undergoing clear corneal phacoemulsification were enrolled in this single surgeon, prospective, double-masked, controlled trial. Patients were randomized into two groups, receiving either intracameral unpreserved lignocaine 1% or placebo (balanced salt solution). Systolic and diastolic blood pressure, pulse rate and peripheral oxygen saturation were recorded preoperatively and during phacoemulsification. The level of intraoperative pain was assessed by a numerical analogue scale ranging between 0 (no pain) and 10 (unbearable pain). Data were compared by Student t-test. RESULTS: Sixty-seven cases received lignocaine (group 1) and 68 received placebo (group 2). Average age was 75 years and 74 years, respectively. Systolic blood pressure, pulse rate, oxygen saturation and pain score showed no statistical difference (P = 0.241, 0.542, 0.712, 0.237, respectively). Diastolic blood pressure showed a weakly significant change (P = 0.023). Patients reported minimal discomfort during surgery in both groups. CONCLUSION: This study found no additional benefit of intracameral unpreserved lignocaine when performing routine clear corneal phacoemulsification under topical anaesthesia. PMID- 11885790 TI - Intraepidermal carcinoma of the eyelid. AB - PURPOSE: To examine the clinical and pathological features and treatment outcomes of cutaneous squamous intraepidermal carcinoma of the eyelid in order to determine its behaviour, its relationship to squamous cell carcinoma and appropriate management. METHODS: All patients from the practice of one of the authors with a histological diagnosis of squamous intraepidermal carcinoma were included. Retrospective chart review was performed, and pathology re-examined. All patients were recalled for examination. Histological diagnosis of periocular intraepidermal carcinoma, lesion characteristics, outcome of surgical excision, local recurrence, and occurrence of metastasis were assessed as main outcome measures. RESULTS: Thirty-one white, usually fair-haired patients had a total of 37 lesions. Most had a history of occupational or recreational sun exposure. Other predisposing factors included exposure to arsenic, petroleum by-products and epidermodysplasia verruciformis. All except one had other solar keratoses and non-melanoma skin cancers. All had surgical excision. Six cases (16%) had evidence of progression to squamous cell carcinoma. Seven cases (18%) had local recurrence successfully treated by further local excision. No patient had perineural spread or distant metastasis. CONCLUSIONS: This lesion, which serves as a marker for severe actinic damage, may masquerade as chronic blepharitis. Early diagnosis prior to dermal invasion is important. Complete excision is the recommended treatment. Adjunctive treatment with topical 5-fluorouracil may be appropriate in some circumstances. Long-term follow up is mandatory. PMID- 11885791 TI - Relationship between the concentration of copper and iron in the aqueous humour and intraocular pressure in rabbits treated with topical steroids. AB - PURPOSE: To evaluate the concentration of copper and iron in the aqueous humour of steroid-treated eyes, particularly to study the concentration of these metals in relation to steroid-induced increases in intraocular pressure (IOP). METHODS: Adult rabbits of both sexes were selected in order to study the effect of steroids on the concentrations of copper and iron in the aqueous humour and on IOP. The rabbits were acclimatised for 2 weeks prior to the instillation of various drugs into the eyes. Then a steroid (dexamethasone, betamethasone or fluoromethalone) was instilled in both eyes of the rabbits, for about 1 month. Intraocular pressure was measured twice a week. When IOP was significantly increased, the animals were killed. The aqueous humour was collected and analysed for copper and iron using atomic absorption spectrophotometry coupled with graphite fumace. RESULTS: After about 30 days of steroid treatment the mean (+/- SD) IOP in dexamethasone, betamethasone and fluoromethalone treated groups was 17.5 (+/- 4.81) mmHg, 18.48 (+/- 4.5) mmHg and 21.8 (+/- 5.7) mmHg, respectively. These values were significantly higher compared to the control group where the mean IOP was 11.6 (+/- 2.2) mmHg. The concentration of copper in the aqueous humour of steroid-treated rabbits was significantly lower (P < 0.001) compared to the control group. However, the concentration of iron was not significantly different between the control and steroid treated rabbits. CONCLUSION: A greater increase in IOP was observed in the fluoromethalone-treated group compared to the dexamethasone and betamethasone-treated groups, but the difference was not significant. The lower concentrations of copper in aqueous humour in steroid treated eyes may play an important role in the maintenance of IOP. The concentration of iron was not significantly different compared to the control group. These results may help to explain the role of these metals in the pathogenesis of open angle glaucoma. PMID- 11885792 TI - Visual field assessment and the Austroads driving standard. AB - PURPOSE: To compare the conventional (Humphrey 24-2) automated visual field testing with the Goldmann standard visual field test for driving, and to predict how many patients with glaucoma may not meet the Australian driving standard with respect to visual fields. METHODS: Four patients (retinitis pigmentosa, glaucoma or vigabatrin treatment) with marked visual field defects as determined by uniocular static computerized perimetry (conventional testing) were re-evaluated with binocular kinetic Goldmann IV4e target field test (Australian driving standard). A series of 48 consecutive patients seen by the Glaucoma Inheritance Study in Tasmania were assessed with both static computerized perimetry and the Goldmann IV4e target test. RESULTS: The four patients with severe visual field defects (on computerized perimetry) were found to meet the driving standard on the binocular Goldmann IV4e target test. On computerized perimetry, 15 of 48 patients from the Glaucoma Inheritance Study in Tasmania were found to have visual field defects of sufficient severity that they may not meet the driving standard. However, only five of these patients failed the driving standard for visual fields, two of whom were still driving. CONCLUSIONS: Patients with severe field defects on conventional uniocular automated perimetry may still meet the Goldmann standard visual field test for driving. Approximately 30% of glaucoma patients would have visual field loss shown on Humphrey 24-2 test of a severity that requires further testing to determine if they meet the driving standard. Ten per cent of glaucoma patients tested did not meet the driving standard for visual fields. PMID- 11885793 TI - Development and surgical implantation of a vision prosthesis model into the ovine eye. AB - PURPOSE: A surgical technique was designed and tested to enable the implantation of an intraocular electrical retinal stimulator. METHOD: An inoperative perspex and silicone model was constructed to closely resemble the anticipated properties of the proposed visual prosthesis. The animal model chosen for these experiments was the sheep, because the dimensions of its ocular anatomy are approximately 30% larger than the human's, being otherwise grossly similar. The surgical method involved transplanar port-hole lensectomy and vitrectomy, insertion of the model implant through a limbal incision, and fixation of the perspex subunit close to the location of the native crystalline lens, by way of trans-pars plana fixation sutures. Adequate pre-retinal positioning of the implant's silicone extension was obtained by way of its inherent elastic recoil. RESULTS: The procedure was performed without macroscopic evidence of undue surgical trauma. CONCLUSION: Although further long-term experiments are required to fully assess the surgical procedure and biocompatibility of the implant, intraoperative assessment and postmortem computed tomographic imaging of the globe has confirmed the successful intraocular positioning and fixation of the implant. PMID- 11885794 TI - Orbital teratoma: late presentation with normal vision. AB - Orbital teratoma is a rare, rapidly growing tumour that usually presents with congenital proptosis. Visual outcome is usually poor. A case is described of orbital teratoma presenting in an 18-month-old child as an inferior orbital mass with normal vision and intermittent hypertropia. Computed tomography demonstrated a cystic mass containing a tooth adjacent to the inferior orbital fissure. Histology revealed tissues derived from all three germ cell layers. The tumour was removed with preservation of vision and resolution of the strabismus. Late presentation of an orbital teratoma with a good visual outcome is exceptional. PMID- 11885795 TI - Acute dacryocystitis presenting as an orbital abscess. AB - Acute dacryocystitis usually presents as a preseptal infection, but can uncommonly be associated with orbital cellulitis. Orbital abscess formation is, however, very rare. The case is presented of a 60-year-old woman with an extraconal abscess secondary to acute dacryocystitis. The clinical, radiological and intraoperative findings are discussed. PMID- 11885796 TI - Post-traumatic Scedosporium inflatum endophthalmitis. AB - This is the first documented case of post-traumatic Scedosporium inflatum endophthalmitis and only the second of S. inflatum endophthalmitis occurring in a non-immunocompromised individual, to the authors' knowledge. A case is reported of a 57-year-old woman who, while chopping wood, had a wood chip hit her in the right eye. This caused a penetrating corneal injury with uveal prolapse and damage to the crystalline lens. There were also vitreous and suprachoroidal haemorrhages. No detectable intraocular foreign material was retained. The clinical manifestation of infection was delayed, but once established, it was very destructive. The initially indolent endophthalmitis eventually led to loss of all light perception and panophthalmitis which required enucleation. The responsible strain of S. inflatum was found to be resistant to all antifungal medication in vitro. PMID- 11885797 TI - Infectious keratitis in orthokeratology. AB - Orthokeratology is a method of changing refraction in myopic patients by using rigid contact lenses to reduce the curvature of the cornea. This treatment was in use in the two cases of corneal ulcer described in this paper and appears to have contributed to the development of their disease. As with extended wear contact lenses, patients undergoing orthokeratology treatment are frequently advised to wear the orthokeratology lenses overnight increasing the risk of corneal ulceration and infection. Patients should be adequately warned of the associated risks and advised that any envisaged benefits of the procedure are temporary. PMID- 11885799 TI - Posterior subtenon injection of corticosteroids using polytetrafluoroethylene (PTFE) intravenous cannula. AB - Injection of corticosteroids into the posterior subtenon space is a well established and highly effective modality in the treatment of intermediate uveitis. The conventional technique of posterior subtenon injection involves the use of a sharp tipped 26-gauge, 5/8 inch needle that must be inserted up to its hub to obtain adequate placement of the drug into the posterior subtenon space. With this technique the risk of perforation of the globe, although minimal, remains a potential complication. Herein is described a new technique for injection of corticosteroids into the posterior subtenon space using an intravenous cannula made of polytetrafluoroethylene (PFTE) that allows safer delivery of the drug into the posterior subtenon space. PMID- 11885798 TI - Combined coliform and anaerobic infection of the lacrimal sac. AB - A case is reported of combined coliform and anaerobic bacterial infection of the lacrimal sac, a condition of which there is only one other published case report. In addition, a literature review is presented of the bacteriology of acute dacryocystitis as it applies to this case. Recommendations for the microbiological investigation and management of acute dacryocystitis are made. PMID- 11885800 TI - Which procedure, which eye? PMID- 11885801 TI - Health status of Lebanese ophthalmologists. PMID- 11885802 TI - Comparison of the frequency doubling technology screening algorithm and the Humphrey 24-2 SITA-FAST in a large eye screening. AB - PURPOSE: To compare the Frequency Doubling Technology (FDT) C20-1 screening algorithm and the Humphrey Field Analyser II (HFA) 24-2 SITA-FAST in a large eye screening. METHODS: In a non-randomized, prospective, free eye screening, the FDT Screening Protocol (C20-1 Screening Algorithm) was administered to 574 attendees (422 men and 152 women, average age 64, range 17-89 years) of the 1998 Veterans of Foreign Wars (VFW) Convention in San Antonio, Texas. Individuals who failed the FDT (two or more misses out of 17 locations) immediately underwent white-on white threshold visual field perimetry (HFA 24-2, SITA-FAST). Humphrey visual field analysis included STATPAC and masked evaluations by three glaucoma specialists. RESULTS: Approximately one-tenth of the VFW conference attendees voluntarily presented themselves for screening. Among these 574 volunteers, 69 (12%) failed the FDT and underwent HFA analysis. Eighty-one per cent (56/69) of these FDT failures had abnormal HFA Glaucoma Hemifield Tests. Eighty-eight per cent (61/69) were judged to have nerve fibre type visual field loss on HFA by at least two of three masked examiners. A positive correlation existed between the number of FDT locations missed and the HFA mean deviation (r = 0.5, P = 0.0001). A similar association was observed when FDT and HFA results were analysed by quadrant (r = 0.5, P < 0.0001). CONCLUSION: There was a low false positive rate and a good positive predictive value comparing the FDT screening algorithm to the HFA 24-2 SITA-FAST in this study. This supports the potential use of FDT as an economical screening device. PMID- 11885804 TI - Immunohistochemical localization of the somatostatin receptor subtype 2A in the rat adrenal gland. AB - The immunohistochemical localization of the somatostatin receptor subtype sst2A was investigated in the rat adrenal gland using SS-800 polyclonal antibody. The sst2A immunopositivity was found in all adrenocortical zones and in adrenal medulla, the reaction being slightly more intense in zona glomerulosa and medulla. The administration of the potent agonist of sst2 receptors - octreotide resulted in the enhancement of the immunopositivity in zona glomerulosa and medulla, whereas chronic exposure of the rats to diethylstilbestrol led to enhancement of the immunopositivity in zona glomerulosa and in the external part of zona fasciculata. PMID- 11885803 TI - Noradrenergic and peptidergic innervation of the mammary gland in the immature pig. AB - The presence and pattern of coexistence of some biologically active substances in nerve fibres supplying the mammary gland in the immature pig were studied using immunohistochemical methods. The substances studied included: protein gene product 9.5 (PGP), tyrosine hydroxylase (TH), somatostatin (SOM), neuropeptide Y (NPY), galanin (GAL), calcitonin gene-related peptide (CGRP) and substance P (SP). The mammary gland was found to be richly supplied by PGP-immunoreactive (PGP-IR) nerve fibres that surrounded blood vessels, bundles of smooth muscle cells and lactiferous ducts. The vast majority of these nerves also displayed immunoreactivity to TH. Immunoreactivity to SOM was observed in a moderate number of nerve fibres which were associated with smooth muscles of the nipple and blood vessels. Immunoreactivity to NPY occurred in many nerve fibres associated with blood vessels and in single nerves supplying smooth muscle cells. Solitary GAL-IR axons supplied mostly blood vessels. Many CGRP-IR nerve fibres were associated with both blood vessels and smooth muscles. SP-IR nerve fibres richly supplied blood vessels only. The colocalization study revealed that SOM, NPY and GAL partly colocalized with TH in nerve fibres supplying the porcine mammary gland. PMID- 11885805 TI - Vasopressinergic innervation of the pig pineal gland. AB - An immunohistochemical study of the pineal gland of the domestic pig was carried out using the antisera raised against vasopressin (VP). The pineal glands were taken from the newborn, 21-day- and 7-month old female pigs. The pig pineal gland is moderately innervated by VP-immunoreactive nerve fibers. They run from the habenular commissure into the connective tissue septa and further into the pineal parenchyma. In the subependymal tissue as well as in the connective tissue septa, the fibers are smooth or with small varicosities and in the parenchyma with large ones. The obtained results point to extrapineal and extraepithalamic source of the fibers. The density of VP-immunoreactive fibers in the pineal gland of 7 month old pigs is higher than in the younger animals. PMID- 11885806 TI - The effect of heat shock, cisplatin, etoposide and quercetin on Hsp27 expression in human normal and tumour cells. AB - Heat shock protein 27 (Hsp27) belongs to the group of proteins called molecular chaperones protecting normal and tumour cells against many stressors such as hyperthermia, several commonly used chemotherapeutics as well as other apoptotic stimuli. Our study was designed to determine whether heat shock and drugs like cisplatin, etoposide and quercetin have an effect on the expression of heat shock protein 27 in tumour cells such as: HeLa (cervical cancer), Hep-2 (larynx cancer), A549 (lung cancer) and also in normal human skin fibroblasts (HSF) cultured in two-dimensional (2D) and three-dimensional (3D) conditions. Our results indicate that Hsp27 expression is drug- and cell-type specific and depends on the culture model as well. HeLa, Hep-2 and HSF cell cultured in 3D model appeared to be more resistant to stimulatory or inhibitory effects of the applied drugs than cells cultured in 2D model. Only A549 cells cultured in 3D culture model appeared to be more susceptible and reacted to drugs and heat treatment with increased Hsp27 expression. PMID- 11885807 TI - Immunohistochemical assay of p53, cyclin D1, c-erbB2, EGFr and Ki-67 proteins in HPV-positive and HPV-negative cervical cancers. AB - The purpose of the present study was to analyse clinical correlation between HPV type 16 and 18 infection, expression of p53, cyclin D1, Ki-67, c-erbB2 and EGFr gene products in cervical cancer cells as well as their nuclear ploidy. The morphological parameters evaluated, such as differentiation of carcinomas, vascular invasion, ploidy and expression of oncogenic proteins, indicate the increased biological malignancy of HPV 16/18-positive carcinomas. The majority of them were poorly differentiated, revealed significantly higher frequence of vascular invasion (p<0.05), were more frequently aneuploid and showed overexpression of cyclin D1. The comparison of the data obtained with the mortality rate of the patients suggests that the overexpression of EGFr and moderate expression of Ki-67 seem to be unfavorable prognostic factors, regardless of the presence of HPV 16/18. PMID- 11885808 TI - rDNA amplification in previtellogenic and vitellogenic oocytes of symphylans (Arthropoda, Myriapoda). AB - Tube-shaped ovaries of symphylans house numerous developing oocytes that are accompanied by somatic follicular cells. Oocyte nuclei (germinal vesicles) are relatively large and ovoid. During early previtellogenesis they contain compact spherical bodies and lampbrush chromosomes immersed in a translucent karyoplasm. Fluorescent labeling with DAPI and propidium iodide has revealed the presence of both DNA and RNA in the spherical bodies. As previtellogenesis advances, small RNA- and AgNOR-positive nucleoli bud off from these bodies. Full-grown nucleoli consist of coarse-granular material and comprise electron-transparent vacuoles. Our results suggest that in symphylan germinal vesicles amplification of rDNA genes takes place, and that the spherical bodies represent accumulations of extrachromosomal rDNA (rDNA bodies) after commencement of transcriptional activity. PMID- 11885809 TI - Nucleolar activity of germ cells in polytrophic ovaries of crane flies (Diptera: Tipulidae). Ultrastructural and cytochemical studies. AB - Ultrastructural and histochemical studies confirmed extrachromosomal amplification of rDNA in nuclei of the oocyte and one of its sibling nurse cells during early stages of oogenesis in Tipula spp. Decondensation of the extra DNA body in the oocyte nucleus coincides with the appearance of multiple nucleoli. In contrast, the amplified copies of ribosomal genes in the nurse cell nucleus remain condensed (i.e. transcriptionally inactive). Roughly of the same size, all nurse cells look almost identical. Their nuclei are spherical and contain single, prominent nucleoli, clumps of chromatin and accumulations of granular material. The cytoplasm is packed with free ribosomes, while in close vicinity of the nuclear envelope many islets of fine granular nuage material can be found. These data indicate that the nurse cells in crane fly ovaries are synthetically active, i.e. contribute to the overall production of ribosomes and their final accumulation in the oocyte. The invariable volume of the nurse cells throughout oogenesis may therefore result from the differences in the dynamics of transcriptional activity and transport of ribosomes, rather than indicate their low synthetic activity. PMID- 11885811 TI - Influence of 4-day long treatment with vasoactive intestinal peptide on ultrastructure and function of the rat pinealocytes in organ culture. AB - Vasoactive intestinal peptide (VIP) is one of neuropeptides involved in the regulation of the pineal gland function. The acute treatment of rat pinealocytes with VIP caused changes in their biochemical parameters. The present study concerns the effects of the chronic treatment with VIP on ultrastructure and function of the rat pinealocytes in organ culture. The pineals of adult male rats were assigned to one of three groups and placed in organ culture for four consecutive days. The pineals of the first group were incubated in the control medium, the pineals of the second group--12 hrs in control medium and 12 hrs in medium with 1 microM VIP (between 20.00 and 8.00) during each day, the pineals of the third group--24 hrs per day in medium with 1 microM VIP. The melatonin concentration was measured using RIA and activity of enzymes using radiochemical methods. Point count method was used in quantitative ultrastructural analysis. Both modes of chronic treatment with VIP increased significantly the level of melatonin secretion during four days of the culture and the content of this hormone in the pineal explants at the end of the experiment. Treatment with the neuropeptide for 12 hrs and 24 hrs per day elevated also the activity of arylalkylamine N-acetyltransferase and hydroxyindole-O-methyltransferase. On the other hand, VIP had no effect on the activity of arylamine-N-acetyltransferase. VIP increased the relative volume of rough endoplasmic reticulum, Golgi apparatus and mitochondria and did not influence the relative volume of lysosomes and lipid droplets as well as the numerical density of dense core vesicles in the examined rat pinealocytes. The obtained results indicate stimulatory effect of chronic treatment with VIP on the synthesis and secretion of melatonin in the rat pinealocytes in vitro. The results of morphological study are in agreement with the obtained biochemical data and point to the increase in secretory and metabolic activity of the rat pinealocytes in response to VIP. PMID- 11885810 TI - Induction of premature mitosis in root meristem cells of Vicia faba and Pisum sativum by various agents is correlated with an increased level of protein phosphorylation. AB - The intra-S-phase checkpoint response to hydroxyurea (HU)-mediated arrest of DNA replication was analysed in root meristems of two legumes, Pisum sativum and Vicia faba. The obtained results suggest that a molecular signal which invokes mechanisms allowing the cells to override the S-M dependency control system may be generated by caffeine (CF) and a number of alternative, yet related chemical agents, benzyl-6-aminopurine (BAP), 2-aminopurine (2-AP), and 6 dimethylaminopurine (DMAP). A variety of aberrant mitotic divisions included chromosomal breaks and gaps, lost and lagging chromatids and chromosomes, acentric fragments, chromosome bridges and micronuclei. Furthermore, similar effects induced by sodium vanadate, an inhibitor of protein phosphatases, extend the number of inhibitors capable of inducing premature chromosome condensation (PCC) in root meristem cells, as well as the range of possible regulatory pathways leading to the transition from S-phase arrest towards abnormal mitosis. Until preprophase, FITC-conjugated monoclonal antibodies (alpha-Y(a)b-FITC) that specifically recognize phosphorylated form of threonine indicate no evident cell cycle-dependent changes in an overall phosphorylation status of root meristem cells in the control plants. Irrespective of the stage of interphase, alpha Y(p)ab-FITC was localized basically in the cytoplasm, whereas nuclear staining was considerably weaker, with a significant fluorescence confined merely to nucleolar regions. The intensity of alpha-Y(p)ab-FITC staining in HU/CF-treated seedlings was found higher than that in the control plants (with the exception of G2 cells), suggesting a general increase in the level of protein phosphorylation, a physiological response mediated probably by an enhanced activity of the cdc like protein kinase(s). PMID- 11885812 TI - Attachment disruptions in seriously emotionally disturbed children: implications for treatment. AB - In this paper, we consider the effect of attachment disruptions on severe adjustment problems in school-age boys. Three groups of 9-11-year-old boys were sampled based on their degree of risk for adjustment difficulties: (1) boys in regular classrooms, (2) boys in regular classrooms who are at risk due to poverty, and (3) boys who have been placed in special education classrooms as a result of serious emotional disturbance (SED). Attachment disruptions were categorized according to the severity of major separations from the biological mother. SED children experienced significantly more severe disruptions of their relationships with their biological mothers and fathers than either the high-risk or comparison boys. Teachers' ratings indicated that both the high-risk and SED boys experienced more externalizing symptoms than comparison boys in regular classrooms. However, SED children were most clearly discriminated from their high risk and comparison counterparts by higher levels of dissociative symptoms. Regression analyses indicated that children who had experienced maternal attachment disruptions were more likely to show dissociative symptomatology in the classroom setting and were more likely to develop dependent relationships with their teachers after risk group status, child age and family structure were controlled. Implications of these findings for the treatment of SED children are discussed. PMID- 11885813 TI - Attachment representations in adolescence: further evidence from psychiatric residential settings. AB - The Adult Attachment Interview (AAI) has afforded the opportunity to profile the various ways in which people make sense of early experience. While the initial research with the AAI was primarily based on non-clinical populations, this paper extends the growing body of knowledge concerning attachment representations in clinical samples, specifically among severely emotionally disturbed adolescents. The study investigated 39 adolescents resident on five regional adolescent units in the south-east of England. As predicted, the number of adolescents presenting as securely attached was low (n = 4), whilst the incidence of insecure attachment patterns in the sample was high (n = 35). When interviews were rated additionally in terms of lack of resolution, 59% of the sample were unresolved with respect to experiences of trauma or loss. Discussion addresses the possible uses of the AAI in therapeutic interventions for severely disturbed adolescents, which are centrally based on the formation of a secure, safe relationship with a non threatening adult. PMID- 11885814 TI - Current attachment representations of incarcerated offenders varying in degree of psychopathy. AB - The present study sought to examine the current mental representations of early attachment relationships in 24 psychopathic criminal offenders, incarcerated in a forensic psychiatric hospital or a medium-security prison. The participants had been assessed on Hare's Psychopathy Checklist, Revised: Screening Version (PCL-R, sv, 1997) and scored either high or low. They were interviewed with the Main and Goldwyn Adult Attachment Interview (1998) and completed the EMBU, a Swedish self report questionnaire tapping memories of the parent's rearing techniques. The results pointed to an extensive over-representation of individuals who were dismissing of attachment and attachment-related experiences (close to three times as many as in the normal population), no secure individuals, and with the remainder being either unclassifiable or unresolved with regard to severe early abuse/trauma. In addition, an examination of the EMBU data revealed an association between a higher psychopathy score and a family constellation of a rejecting father and an emotionally very warm (idealized) mother. The discussion will focus on the unique discourse of the dismissing individuals and on clinical implications. PMID- 11885815 TI - The adult attachment interview: rating and classification problems posed by non normative samples. AB - Non-normative samples can pose major procedural and coding challenges to interviewers and raters of the Adult Attachment Interview (AAI). With reference to interview transcripts drawn from a population of personality disordered offenders detained in a high-security hospital, specific difficulties are identified and discussed. These difficulties have their roots in three separate but overlapping areas: extreme attachment-related experience; interviewees' psychological or psychiatric state; and factors relating to the context in which the interview is conducted. They raise questions about whether and when the use of the interview should be restricted, the rating rules elaborated and/or the rating system expanded. Suggestions are made as to how some of the difficulties might be addressed. PMID- 11885816 TI - Attachment in mental health institutions: a critical review of assumptions, clinical implications, and research strategies. AB - Attachment is relevant to institutionalized treatment and the therapeutic process in three identifiable ways: (1) patients bring their mental representations of previous and existing attachment relationships to the treatment; (2) attachment is relevant to the extent to which a therapeutic alliance is established and maintained, both in terms of the mental representations of attachment in the patient and in the therapist and how these influence interactive behaviour and expectations in each partner to the therapeutic work; (3) the outcome of the treatment may be related to attachment; for example, when institutional experiences have an enduring impact on attachment representations and the future attachment behaviour of the patient. However, this brief review of attachment concepts reveals that several theoretical, conceptual and empirical questions remain to be answered before evidence-based clinical attachment guidelines can be formulated concerning patient-staff relationships. PMID- 11885817 TI - Attachment in mental health institutions: a commentary. PMID- 11885818 TI - Can attachment theory, and attachment research methodologies, help children and adolescents in mental health institutions? PMID- 11885820 TI - Why women consult with increased vaginal bleeding: a case-control study. AB - BACKGROUND: Many women with heavy periods and irregular bleeding do not consult about them. It has been suggested that some of these symptoms are associated with psychological distress and that this influences consultation behaviour which may account for why some women present with a menstrual disturbance and others with apparently the same problem do not. AIM: To explore the relationship between symptom severity, psychological distress, and the seeking of medical help in primary care among women aged 54 years or less with increased vaginal bleeding. DESIGN OF STUDY: Case control. SETTING: An urban four-partner general practice of 10,000 patients. METHOD: Questionnaires were sent to women who were consulting with new episodes of 'increased vaginal bleeding' and two groups of controls: consulting controls with 'acute respiratory tract infection' (RTI) or 'other illness' as identified by weekly computerised searches, and community controls, selected from the practice age-sex register. RESULTS: Nine hundred and forty three questionnaires were sent out to 108 cases and 835 controls with an 80% response rate. Of these, 60.9% of the cases, 47.0% of the consulting controls, and 39.7% of the community controls were subjects with probable psychological distress on the General Health Questionnaire (chi2 test, P = 0.002). Cases were more likely than community controls to have heavy periods (odds ratio [OR] = 2.86, 95% confidence interval [CI] = 1.53-5.35) and heavy periods interfering with life (OR = 3.69, 95% CI = 2.02-6.75). After controlling for heaviness of periods, cases were still more likely to have psychological distress (OR = 1.80, 95% CI = 1.00-3.24). The same relationships prevailed when comparing cases and consulting controls. CONCLUSION: Interference in life caused by heaviness of periods appears to be a powerful initiator of consultation with increased vaginal bleeding. Perceived heavy periods and psychological disturbance are weaker predictors. Women presenting to primary care with increased vaginal bleeding are more likey to have a psychological disturbance than women from the community or those consulting with another illness. PMID- 11885819 TI - Relationship style between GPs and community mental health teams affects referral rates. AB - BACKGROUND: Community mental health teams (CMHTs) are the established model for supporting patients with serious mental illness in the community. However, up to 25% of those with psychotic disorders are managed solely by primary care teams. Effective management depends upon locally negotiated referral and shared care arrangements between CMHTs and primary care. AIM: To examine whether the style of working relationship between general practices and CMHTs affects the numbers and types of referrals from general practices to CMHTs, taking into account population and practice factors and provision of other mental health services which may influence referral rates. DESIGN OF STUDY: Cross-sectional study. SETTING: All 161 general practices in East London and the City Health Authority. METHOD: Questionnaire survey to all general practices to identify style of relationship. Collection of routinely available referral data to all statutory mental health services over a two-year period. Main outcome measures were number and types of referrals from general practices to CMHTs. RESULTS: The average annual referral rate to the eleven CMHTs in east London is 10 per 1000 adult population annually. The teams show a sixfold variation in rates of referral from all sources. Where good working relationships (a consultation-liaison style) exist between CMHTs and general practice, there are greater numbers of referrals requiring both long and short-term work by CMHTs. Two-stage multivariate models explained 47% of the referral variation between practices. Where primary care based psychologists work with practices there are greater numbers of CMHT referrals, but less use of psychiatric services. CONCLUSION: Shifting to a consultation-liaison relationship should increase rates of referral of patients with serious mental illness, including those who can most benefit from the skills of CMHTs. Increasing the provision of primary care-based psychology might improve practice use of mental health services, reducing avoidable outpatient psychiatric referrals. PMID- 11885821 TI - Inequalities in morbidity and consulting behaviour for socially vulnerable groups. AB - BACKGROUND: The focus of health policy on improving health and reducing inequality for socially vulnerable groups. AIM: To examine self-report of condition-specific morbidity and consultation with the general practitioner (GP) for socially vulnerable groups. DESIGN OF STUDY: Cross-sectional survey using a modified version of the General Practitioner Assessment Survey (GPAS). SETTING: Ten general practices in each of six health authorities. METHOD: A random sample of 200 patients was selected from each practice. The questionnaire elicited information about experience of specific acute and chronic conditions and whether the GP had been consulted. Four sub-samples were selected from the 4493 registered patients who responded to the self-completion questionnaire. They were lone mothers (n = 160), elderly living alone (n = 417), the unemployed (n = 100), and members of ethnic minority groups (n = 316). RESULTS: Logistic regression analyses showed that, after adjustment for age, sex, smoking, and housing tenure, only lone motherhood and ethnic minority group status were consistently and independently associated with poorer health outcomes. Lone motherhood was associated with a higher likelihood of anxiety (odds ratio [OR] = 2.03, 95% confidence interval [CI] = 1.34 to 3.08) and sleep problems (OR = 1.83, 95% CI = 1.18 to 2.83) and ethnic minority group status with a higher likelihood of depression (OR = 2.02, 95% CI = 1.34 to 3.04), diabetes (OR = 4.03, 95% CI = 2.54 to 6.39, migraine (OR = 1.72, 95% CI = 1.26 to 2.35), and minor respiratory symptoms (OR = 1.75, 95% CI = 1.33 to 2.29). Ethnic minority group status was the only source of social vulnerability that was independently associated with a higher likelihood of GP consultation, particularly for episodes of illness such as backache (OR = 3.28, 95% CI = 2.06 to 5.21), indigestion (OR = 2.94, 95% CI = 1.53 to 5.65), migraine (OR = 3.22, 95% CI = 1.75 to 5.93), minor respiratory symptoms (OR = 3.53, 95% CI = 2.26 to 5.50) and sleep problems (OR = 4.72, 95% CI = 2.56 to 8.71). CONCLUSIONS: Social vulnerability can be a risk factor for poorer health, but this is dependent on the source of vulnerability and is condition-specific. No association was found between inequity in the utilisation of primary care and social vulnerability. The propensity for members of ethnic minority groups to consult more than white people, particularly for acute conditions, requires further exploration. PMID- 11885822 TI - Comparison of the smoking behaviour and attitudes of smokers who attribute respiratory symptoms to smoking with those who do not. AB - General practitioners' (GPs') advice against smoking helps smokers to stop; unfortunately, GPs cannot predict which patients will quit following advice. This postal questionnaire survey suggests that where smokers attribute their respiratory symptoms to smoking, they are eight times (95% confidence interval [CI] = 3.0-23.3) more likely to believe that their health will improve if they stop smoking and six times (95% CI = 1.4-23.3) more likely to intend to stop smoking. PMID- 11885823 TI - Toys are a potential source of cross-infection in general practitioners' waiting rooms. AB - The waiting rooms of general practitioners' surgeries usually have toys provided for children. The level of contamination of these toys and the effectiveness of toy decontamination was investigated in this study. Hard toys from general practitioners' waiting rooms had relatively low levels of contamination, with only 13.5% of toys showing any coliform counts. There were no hard toys with heavy contamination by coliforms or other bacteria. Soft toys were far more likely to be contaminated, with 20% of toys showing moderate to heavy coliform contamination and 90% showing moderate to heavy bacterial contamination. Many waiting-room toys are not cleaned routinely. Soft toys are hard to disinfect and tend to rapidly become recontaminated after cleaning. Conversely, hard toys can be cleaned and disinfected easily. Soft toys in general practitioners' waiting rooms pose an infectious risk and it is therefore recommended that soft toys are unsuitable for doctors' waiting rooms. PMID- 11885824 TI - The abolition of the GP fundholding scheme: a lesson in evidence-based policy making. AB - The general practitioner (GP) fundholding scheme was introduced as part of the Conservative governments 1991 National Health Service reforms and abolished by the Labour government in 1998. This paper contends that the scheme was introduced and abolished without policy-makers having any valid evidence of its effects. In particular, it focuses on the salient features of the decision to abolish. These were: (a) that it was not based on evidence; (b) that it came relatively soon after the introduction of the scheme; and (c) the GP fundholding scheme was voluntary and increasing numbers of GPs were being recruited. The overtly political nature of the introduction of GP fundholding is already well documented and is important in understanding the lack of evidence involved in the development of the fundholding scheme. PMID- 11885826 TI - Left ventricular dysfunction in the diabetic population. PMID- 11885827 TI - Predictive value of asthma medication to identify asthma sufferers. PMID- 11885828 TI - Melleril: gone forever! PMID- 11885825 TI - A combination of systematic review and clinicians' beliefs in interventions for subacromial pain. AB - The aim of the study is to determine which treatments for patients with subacromial pain are trusted by general practitioners (GPs) and physiotherapists, and to compare trusted treatments with evidence from a systematic critical review of the scientific literature. A two-step process was used: a questionnaire (written case simulation) and a systematic critical review. The questionnaire was mailed to 188 GPs and 71 physiotherapists in Sweden. The total response rate was 72% (186/259). The following treatments were trusted, ergonomics/adjustments at work, corticosteroids, non-steroidal anti-inflammatory drugs, movement exercises, acupuncture, ultrasound therapy, strengthening exercises, stretching, transcutaneous electric nerve stimulation, and superficial heat or ice therapy. The review, including efficacy studies for the treatments found to be trusted, was conducted using the CINAHL, EMBASE and MEDLINE databases. Evidence for efficacy was recorded in relation to methodological quality and to diagnostic criteria that labelled participants as having subacromial pain or a non-specific shoulder disorder. Forty studies were included. The methodological quality varied and only one treatment had definitive evidence for efficacy for non-specific patients, namely injection of corticosteroids. The trust in corticosteroids, injected in the subacromial bursa, was supported by definitive evidence for short term efficacy. Acupuncture had tentative evidence for short-term efficacy in patients with subacromial pain. Ultrasound therapy was ineffective for subacromial pain. This is supported by tentative evidence and, together with earlier reviews, this questions both the trust in the treatment and its use. The clinicians' trust in treatments had a weak association with available scientific evidence. PMID- 11885829 TI - Selling drugs. PMID- 11885830 TI - Selling drugs to doctors--it's marketing, not education. PMID- 11885832 TI - Developments in the provision of primary health care for homeless people. PMID- 11885831 TI - Selling drugs to the public--should the UK follow the example of the US? PMID- 11885834 TI - Hickman catheter complications in a haematology unit, 1996-98. PMID- 11885833 TI - A pragmatic randomised controlled trial of a prompt and reminder card in the care of people with epilepsy. AB - BACKGROUND: The quality of epilepsy care has often been noted to be poor and fragmentary. Most people with epilepsy are solely under the care of their general practitioner (GP). Many patients report medication side-effects and poor seizure control. Most GPs accept responsibility for epilepsy care; however, many report problems with knowledge of epilepsy and nearly all support guidance on epilepsy management. AIM: To determine whether a GP-completed prompt and reminder card is effective in improving the quality of epilepsy care when used opportunistically. DESIGN OF STUDY: Primary care-based pragmatic cluster-randomised controlled trial. SETTING: People with active epilepsy (n = 1275) from 82 practices. METHOD: Practices were randomly categorised as 'control', 'doctor-held card' (card in patient records), or 'patient-held card' practices. RESULTS: Compared with control practices, recording of seizure frequency was significantly increased in doctor-held card practices (57.4% versus 42.8%, P = 0.003) but not in patient held card practices (44.6% versus 42.8%). No differences were found in the proportion of seizure-free patients (doctor-held card [56.0%] versus control [51.5%]; patient-held card [58.1%] versus control [51.5%]) or in the proportion on monotherapy. Patients in both intervention groups reported more medication related side-effects and patients in doctor-held card practices were less satisfied with information provision about epilepsy. Participating GPs found the card useful. The doctor-held card was retrieved and completed more often than the patient-held card. CONCLUSIONS: A doctor-held prompt and reminder card is effective in improving the recording of key clinical information for people with epilepsy, is felt to be useful by GPs, and is completed more often than a patient held card. However it does not improve outcomes and may result in less patient centred care. PMID- 11885836 TI - Schwannoma as a cause of Pancoast's syndrome. PMID- 11885835 TI - Medication use in hospitalized nonagenarians. PMID- 11885837 TI - The clinical basis for the management of coronary artery disease. PMID- 11885838 TI - Cyclophosphamide-induced transitional cell carcinoma of bladder in lupus nephritis. PMID- 11885839 TI - Late onset systemic lupus erythematosus with elevated CA125 and gastrointestinal ischaemia. PMID- 11885840 TI - Slow release nifedipine for patients with sphincter of Oddi dyskinesia: results of a pilot study. PMID- 11885841 TI - Inappropriate thyroid-stimulating hormone secretion. PMID- 11885842 TI - Anchored by a slipping ANCA? PMID- 11885843 TI - Can pre-emptive analgesia reduce pain experienced after liver biopsy? PMID- 11885844 TI - The chronic obstructive pulmonary disease strategy. PMID- 11885845 TI - Evaluating Australia's National Medicines Policy using geographical mapping. AB - BACKGROUND: There has been a proliferation of quality use of medicines activities in Australia since the 1990s. However, knowledge of the nature and extent of these activities was lacking. A mechanism was required to map the activities to enable their coordination. AIMS: To develop a geographical mapping facility as an evaluative tool to assist the planning and implementation of Australia's policy on the quality use of medicines. METHODS: A web-based database incorporating geographical mapping software was developed. Quality use of medicines projects implemented across the country was identified from project listings funded by the Quality Use of Medicines Evaluation Program, the National Health and Medical Research Council, Mental Health Strategy, Rural Health Support, Education and Training Program, the Healthy Seniors Initiative, the General Practice Evaluation Program and the Drug Utilisation Evaluation Network. In addition, projects were identified through direct mail to persons working in the field. RESULTS: The Quality Use of Medicines Mapping Project (QUMMP) was developed, providing a Web based database that can be continuously updated. This database showed the distribution of quality use of medicines activities by: (i) geographical region, (ii) project type, (iii) target group, (iv) stakeholder involvement, (v) funding body and (vi) evaluation method. At September 2001, the database included 901 projects. Sixty-two per cent of projects had been conducted in Australian capital cities, where approximately 63% of the population reside. Distribution of projects varied between States. In Western Australia and Queensland, 36 and 73 projects had been conducted, respectively, representing approximately two projects per 100,000 people. By comparison, in South Australia and Tasmania approximately seven projects per 100,000 people were recorded, with six per 100,000 people in Victoria and three per 100,000 people in New South Wales. Rural and remote areas of the country had more limited project activity. CONCLUSIONS: The mapping of projects by geographical location enabled easy identification of high and low activity areas. Analysis of the types of projects undertaken in each region enabled identification of target groups that had not been involved or services that had not yet been developed. This served as a powerful tool for policy planning and implementation and will be used to support the continued implementation of Australia's policy on the quality use of medicines. PMID- 11885846 TI - Review of the effect of the dosing interval for inhaled corticosteroids in asthma control. AB - BACKGROUND: Asthma is recognized as an inflammatory disease of the airways and treatment includes anti-inflammatory agents such as corticosteroids. Inhaled corticosteroids (ICS) are widely prescribed for long-term prophylaxis, yet their optimal dosing interval is not clear. AIMS: To determine whether the dosing interval of ICS affects asthma control. METHODS: We performed an electronic search of the literature to identify studies on the dosing interval of ICS in asthmatic subjects. Data were extracted from suitable studies by two independent researchers and, where possible, a meta-analysis performed. RESULTS: A total of 4,267 titles were retrieved, of which 13 met inclusion and exclusion criteria and 11 had extractable data. There were no significant differences between outcomes for: (i) once daily vs twice daily administration (7 trials, 810 subjects), (ii) once daily vs four times daily administration (2 trials, 68 subjects) and (iii) twice daily vs four times daily administration (4 studies, 111 subjects). There was a variety of outcomes used to assess differences between dosing intervals. These included symptom scores, lung function, use of rescue medication and adverse drug effects. The number of subjects that could be included in the statistical analysis of any of such outcomes was small, much smaller than the total sample size. CONCLUSIONS: There was no significant difference in measures of asthma control between the assessed dosing intervals of ICS. Current evidence indicates that single daily administration of ICS produces equivalent asthma control to multiple daily administration. PMID- 11885847 TI - Resection of residual pulmonary masses after chemotherapy in patients with metastatic non-seminomatous germ cell tumours. AB - BACKGROUND: Resection of residual post-chemotherapy pulmonary masses in patients with non-seminomatous germ cell tumours gives therapeutic benefit and prognostic information. AIM: This study was undertaken to review the experience of this intervention in a single teaching hospital. METHODS: The Germ Cell Database of the Sydney Cancer Centre was searched for all patients who had undergone excision of pulmonary metastases. These patient records were subsequently reviewed. RESULTS: Between 1976 and 1999, 15 patients underwent a combined total of 19 thoracotomies for resection of residual tumour mass after cisplatin-based chemotherapy. The primary tumour histology included mature teratoma in 47% (7 of 15) of patients. Prior to chemotherapy, 73% (11 of 15) of patients had elevated serum levels of alpha-fetoprotein (median 180 ng/mL) and 60% (9 of 15) of patients had elevated beta-human chorionic gonadotropin (median 672 IU/L). The median length of hospital stay related to thoracotomy was 7 days. There were two surgical complications, a prolonged air leak and a residual pleural effusion. Pathology of residual pulmonary masses revealed necrosis alone in 37% (7 of 19) of procedures, mature teratoma alone in 32% (6 of 19) of procedures and viable tumour in 32% (6 of 19) of procedures. Of those with viable tumour, three achieved long-term complete response (CR), two died of progressive disease (PD) and one is alive with PD. Of those with teratoma, two achieved CR and one relapsed. The long-term CR rate was 80% (12 of 15 patients). The median follow up was 10 years (range 0.75-17.5 years). Four patients died, two of PD and two of cardiovascular disease while in CR. CONCLUSION: At this institution, thoracotomy for residual pulmonary masses was well tolerated, with a high cure rate. PMID- 11885848 TI - Minidose (1 mg) warfarin as prophylaxis for central vein catheter thrombosis. AB - BACKGROUND: Central vein catheters, which are used in the treatment of cancer patients, are prone to thrombotic complications of the catheter or adjacent vein. Previous studies suggest that 1 mg warfarin daily (minidose) can significantly reduce that risk. AIMS: This, study aims to establish whether minidose warfarin could reduce catheter-related thrombosis in adult patients with haematological malignancies. METHODS: Patients were randomly selected to receive warfarin or not. The end-points studied were: (i) occlusion by thrombus, (ii) removal of catheter for other reasons or (iii) 90 days free of thrombus. RESULTS: There was no significant difference in the incidence of catheter thrombosis or venous thrombosis and no significant variation in catheter survival between the study and control groups. CONCLUSIONS: This study found no benefit of the routine use of minidose warfarin for prophylaxis of central vein' catheter thrombosis in patients with haematological malignancies and therefore does not support the routine use of minidose warfarin for prophylaxis in such patients. PMID- 11885849 TI - Ethical, legal and social issues surrounding the human genome project. PMID- 11885850 TI - Health services research: what is it and what does it offer? AB - Over the last 20 years, clinical medicine has witnessed rapid expansion in its underlying evidence base, greater demand for accountability in clinicians' use of limited resources and increasing societal expectation for health care that confers proven benefit at reasonable cost to all eligible recipients. Health services research, also referred to as the clinical evaluative sciences, has grown in response to the need for objective empirical analysis of the modern health system's ability to deliver effective, efficient, equitable and safe care and to further the health and well-being of whole populations. In this article we provide an overview of the aims, methods and outputs of this burgeoning new discipline. PMID- 11885851 TI - Determination of antiepileptic drugs in biological material. AB - Current analytical methodologies applied to the determination of antiepileptic drugs in biological material are reviewed. The role of chromatographic techniques is emphasized. Special attention is focused on new chemical entities as well as current trends such as high-speed liquid chromatographic techniques, hyphenated techniques and electrochromatography techniques. A review with 542 references. PMID- 11885852 TI - External contamination of bovine hair with beta2-agonist compounds: evaluation of decontamination strategies. AB - Hair analysis has shown great potential in the control of illegal use of veterinary drugs such as beta2-agonists. However, it has been shown that hair can be externally contaminated with drugs which can lead to false positive results. Exposure of bovine hair to aqueous solutions of beta2-agonist compounds results in incorporation of these drugs into the hair. Standard hair washing procedures found in the literature: detergent (Tween-20), phosphate buffer or organic solvents (dichloromethane or methanol) cannot eliminate this external contamination. Beta2-agonists can be extracted from hair very efficiently with 0.1 M HCl, the extraction kinetics of externally and endogenously accumulated clenbuterol at room temperature are different which makes it feasible to discriminate between them. Treatment of hair samples with a 0.1 M HCI solution for 2 h at room temperature results in a ratio of clenbuterol content in the wash solution to clenbuterol content in the washed hair equal to or less than 0.25 for samples from treated cattle; whereas this ratio is equal to or higher than 0.70 for externally contaminated samples. The design of the study was intended to resemble the plausible scenario of hair being sampled a short time after external contamination. A similar study to detect external contamination for hair sampled a long time after exposure is in progress. PMID- 11885853 TI - Quantitative liquid chromatography-mass spectrometry determination of isatin in urine using automated on-line extraction. AB - Here we describe a simple, fast and sensitive liquid chromatography/mass spectrometry method with automated on-line extraction to quantify isatin, an endogenous monoamine oxidase, and atrial natriuretic peptide inhibitor, in urine. After derivatisation of isatin to isatinoxime with hydroxylamine hydrochloride and zinc sulfate precipitation, samples were loaded on the extraction column, washed and, after activation of the column-switching valve, backflushed onto the analytical column. Using electrospray ionisation, [M+H]+ ions could be detected in the selected ion monitoring mode. The assay was linear from 5 to 5000 ng/ml (r2>0.99) and analytical recovery was >80%. Inter-assay precision for the quality control samples was less than 3% and inter-assay accuracy was within +/- 5%. PMID- 11885854 TI - Quantitative determination of glufosinate in biological samples by liquid chromatography with ultraviolet detection after p-nitrobenzoyl derivatization. AB - We have established a new HPLC method for derivatizing and quantifying glufosinate (GLUF) in human serum and urine using p-nitrobenzoyl chloride (PNBC). The p-nitrobenzoyl derivative of GLUF (PNB-GLUF) was produced quantitatively over 10 min at room temperature. PNB-GLUF possesses the property of ultraviolet (UV) light absorption with a lambda(max) of 272.8 nm, and was isolated from biological specimens by reversed-phase chromatography using Inertsil Ph-3. In experiments at a UV wavelength of 273 nm, GLUF has a quantitative detection limit of 0.005 microg/ml, and when it was added to both serum and urine to yield concentrations of 0.1-1000 microg/ml, its recovery rate was quite satisfactory: at least 93.8% in all cases. Further, the measured amounts of GLUF in 23 serum samples from patients intoxicated by ingestion of GLUF compared favorably with those obtained by fluorescence derivatization-HPLC using 9-fluorenylmethyl chloroformate (R=0.998). This technique of analysis is, in addition, applicable for Glyphosat, which possesses a chemical structure resembling that of GLUF, and it will be of great use in the determination of these two compounds. PMID- 11885855 TI - Determination of homocysteine thiolactone and homocysteine in cell cultures using high-performance liquid chromatography with fluorescence detection. AB - A sensitive and simple method utilising fluorometric detection for the simultaneous routine monitoring of homocysteine thiolactone (HTL) and homocysteine (Hcy) in biological samples has been developed. Separation relies on isocratic ion-pairing and reversed-phase chromatography while the principle of the detection is that the lactone ring in HTL molecule is cleaved with an alkali to produce Hcy, which reacts with ortho-phthalaldehyde (OPA) in the absence of an added thiol reagent to form a stable fluorescent derivative. The method has a sensitivity of 200 fmol of HTL and 100 fmol for Hcy in the sample. The present method was applied to the determination of HTL and Hcy in Hep G2 cell. PMID- 11885856 TI - Determination of modafinil, modafinil acid and modafinil sulfone in human plasma utilizing liquid-liquid extraction and high-performance liquid chromatography. AB - An assay was developed to determine concentrations of modafinil (dl-2 [(diphenylmethyl)sulfinyl]acetamide; Provigil) and its two major circulating metabolites, modafinil acid and modafinil sulfone, in human plasma. The assay utilized liquid-liquid extraction of the analytes and an internal standard, (phenylthio)acetic acid, from plasma into a mixture of hexane-dichloromethane glacial acetic acid (55:45:2, v/v). The analytes were resolved isocratically on a narrow-bore phenyl column at a mobile phase flow-rate of 0.3 ml/min and were monitored by UV detection at 235 nm. The method reported herein reduces the required sample volume of previously reported methods from 1.00 to 0.200 ml of plasma while lowering the limit of quantification (LOQ). The linear range of the assay was from 0.100 to 20.0 microg/ml for each of the three compounds. PMID- 11885857 TI - Control of propranolol intake by direct chromatographic detection of alpha naphthoxylactic acid in urine. AB - A rapid chromatographic procedure with a C18 column, a mobile phase of 0.15 M sodium dodecyl sulfate (SDS)-10% (v/v) 1-propanol at pH 3 (0.01 M phosphate buffer), and fluorimetric detection, is reported for the control of propranolol (PPL) intake in urine samples, which are injected directly without any other treatment than filtration. The peak of PPL was only observed in samples taken a few hours after ingestion of the drug due to its extensive conjugation and metabolisation. The detection of several unconjugated PPL metabolites was therefore considered: desisopropylpropranolol (DIP), propranolol glycol (PPG), alpha-naphthoxylactic acid (NLT) and alpha-naphthoxyacetic acid (NAC). NLT showed the best characteristics: it eluted at a much shorter retention time than PPL, its concentration in urine samples was greater and it did not present any interference from endogeneous compounds in urine, common drugs or drugs administered in combination with PPL. The limit of quantification, measured as the concentration of analyte providing a relative standard deviation of 20%, was 24 ng/ml, and the day-to-day imprecision was below 4% for concentrations above 200 ng/ml. The procedure allows the routine control of PPL at therapeutic urine levels. Urinary excretion studies showed that the detection of NLT is possible at least up to 20-30 h after oral administration. PMID- 11885858 TI - Ergosteroids. VI. Metabolism of dehydroepiandrosterone by rat liver in vitro: a liquid chromatographic-mass spectrometric study. AB - Because relatively large amounts of dehydroepiandrosterone (DHEA) are required to demonstrate its diverse metabolic effects, it is postulated that this steroid may be converted to more active molecules. To search for the possible receptor recognized hormones. DHEA was incubated with whole rat liver homogenate and metabolite appearances were studied by LC-MS as a function of time to predict the sequence of their formation. An array of metabolites has been resolved, identified and characterized by highly specific and accurate technique of LC-MS, and several of these steroids were analyzed quantitatively. Their identities were established by comparison with pure chemically synthesized compounds and by chemical degradation of isolated fractions. In the present study, we have reasonably established that DHEA was converted to 7alpha-OH-DHEA, 7-oxo-DHEA, and 7beta-OH-DHEA in sequence. These metabolites were further reduced at position 7 and/or 17 to form their respective diols and triols, which were also sulfated at 3beta-position. DHEA and its 7-oxygenated derivatives were also converted to their respective 3beta-sulfate esters. Several of these steroids are being reported for the first time. 16Alpha-hydroxy-DHEA, androst-5-ene 3beta,16alpha,17beta-triol, androst-4-ene-3,17-dione, 11-hydroxy-androst-4-ene 3,17-dione, androst-5-ene-3,17-diol and testosterone were also identified and characterized. In all, 19 metabolites of DHEA are being reported in this extensive study. We have also detected the formation of 12 additional metabolites including several conjugates, which are the subject of current investigation. PMID- 11885859 TI - Identification of a novel selenium metabolite, Se-methyl-N acetylselenohexosamine, in rat urine by high-performance liquid chromatography- inductively coupled plasma mass spectrometry and--electrospray ionization tandem mass spectrometry. AB - The major urinary metabolite of selenium (Se) in rats was identified by HPLC inductively coupled argon plasma mass spectrometry (ICP-MS) and--electrospray tandem mass spectrometry (ESI-MS/MS). As the urine sample was rich in matrices such as sodium chloride and urea, it was partially purified to meet the requirements for ESI-MS. The group of signals corresponding to the Se isotope ratio was detected in both the positive and negative ion modes at m/z 300 ([M+H]+) and 358 ([M+CH3COO]-) for 80Se, respectively. These results suggested that the molecular mass of the Se metabolite was 299 Da for 80Se. The Se metabolite was deduced to contain one methylselenyl group, one acetyl group and at least two hydroxyl groups from the mass spectra of the fragment ions. The spectrum of the Se metabolite was completely identical to that of the synthetic selenosugar, 2-acetamide-1,2-dideoxy-beta-D-glucopyranosyl methylselenide. However, the chromatographic behavior of the Se metabolite was slightly different from that of the synthetic selenosugar. Thus, the major urinary Se metabolite was assigned as a diastereomer of a selenosugar, Se-methyl-N-acetyl-selenohexosamine. PMID- 11885861 TI - Endogenous alkaloids in man. XXXVIII. "Chiral" and "achiral" determination of the neurotoxin TaClo (1-trichloromethyl-1,2,3,4-tetrahydro-beta-carboline) from blood and urine samples by high-performance liquid chromatography--electrospray ionization tandem mass spectrometry. AB - An improved sensitive assay for the determination of the dopaminergic and serotonergic neurotoxin 1-trichloromethyl-1,2,3,4-tetrahydro-beta-carboline (TaClo) is presented, based upon on-line coupling of high-performance liquid chromatography with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS MS). Applying synthetic [D4]TaClo as a fourfold deuterated internal standard, TaClo was detected and reliably quantified as a trace constituent of blood samples (0.5 up to 70 ng g(-1) of clot) obtained from six patients orally treated with the hypnotic chloral hydrate. Unambiguous identification of this tricyclic "endogenous alkaloid" was achieved by selected reaction monitoring (SRM) experiments. The molecular ion peaks of TaClo, m/z 289 (for [35Cl3]TaClo) and m/z 291 (for its [37Cl35Cl2]isotopomer), were both monitored to undergo a retro-Diels Alder fragmentation by loss of a CH2=NH portion (-29 u) as typical of a tetrahydropyrido ring system of tetrahydro-beta-carbolines. Detection of the resulting fragments, m/z 260 and m/z 262, with the expected statistical chlorine isotopic intensities of 100:96 confirmed the identity of the TaClo molecule. In addition, an enantiomer-specific device was developed for TaClo, by employing a chiral reversed-phase HPLC column in combination with circular dichroism (CD) spectroscopy and MS-MS analysis (LC-CD and LC-MS-MS coupling). In a human clot sample, both TaClo enantiomers were found in equimolar concentration (i.e., as a racemate) corroborating a spontaneous, non-enzymatic formation of TaClo from biogenic tryptamine and therapeutically administered chloral. In urine samples of TaClo-treated rats, by contrast, the (S)-antipode was found to predominate, hinting at an enantiomer-differentiating metabolism of the compound. PMID- 11885860 TI - Determination of difloxacin and sarafloxacin in chicken muscle using solid-phase extraction and capillary electrophoresis. AB - This paper describes a method for residue analysis of difloxacin and sarafloxacin in chicken muscle. Clean-up and preconcentration of the samples are effected by solid-phase extraction (C18) and the determination is carried out by capillary electrophoresis using a photodiode array detection system. The method was validated with satisfying results. The calibration graphs are linear for difloxacin and sarafloxacin from 50 to 300 microg/kg. The limit of detection obtained for difloxacin and sarafloxacin are 10 and 25 microg/kg, respectively, which allows the detection of positive muscle samples at the required maximum residue limits of European Union. PMID- 11885862 TI - Screening method for inherited disorders of purine and pyrimidine metabolism by capillary electrophoresis with reversed electroosmotic flow. AB - Capillary electrophoresis with electroosmotic flow reversed by cationic surfactant for diagnosis of purine and pyrimidine inherited enzyme deficiencies is reported. Final separation conditions consist of 45 mM borate, 55 mM N tris[hydroxymethyl]methylglycine, 10 mM tartrate, 1 mM cetyltrimethylammonium bromide and 0.44% tetrabutylammonium hydroxide-2-amino-2-methyl-1,3-propanediol (pH 8.6). Average sensitivity (2.51 microM), reproducibility of migration times (run-to-run C.V. < or = 0.6%, day-to-day C.V. < or = 2.5%), linearity (R2>0.994) and imprecision (mean intra-assay RSD 4.7% and inter-assay RSD 6.6%) of the method are acceptable for diagnostic purposes. Applicability of the method is demonstrated on urine samples from patients with enzymatically proven enzyme deficiencies. PMID- 11885863 TI - Validation of qualitative chromatographic methods: strategy in antidoping control laboratories. AB - An experimental approach for the validation of chromatographic qualitative methods and its application in an antidoping control laboratory is described. The proposed strategy for validation of qualitative methods consists of the verification of selectivity/specificity, limit of detection (LOD), extraction recovery and repeatability (intra-assay precision). A one-day assay protocol, based on the analysis of five blank samples obtained from different sources and four replicates of control samples at two different concentrations of the analytes, has been defined to evaluate the validation parameters. The following evaluation criteria have been applied: absence of interfering substances at the retention time of the analytes in the blank samples to check the selectivity/specificity of the method, the LOD recommended by international sports authorities has to be attained, and for repeatability, the relative standard deviation should be <25% for the low concentration control sample and <15% for the high concentration control sample. Qualitative screening procedures are able to detect a great number of analytes so that extraction and analysis conditions are always a compromise for the different analytes. For this reason, no minimum acceptance criteria have been defined for data of extraction recoveries. The proposed protocol has been used for the validation of the screening and confirmation qualitative methods included in the scope of the accreditation of an antidoping control laboratory according to ISO quality standards. PMID- 11885864 TI - Direct cocktail analysis of drug discovery compounds in pooled plasma samples using liquid chromatography-tandem mass spectrometry. AB - Direct plasma injection technology coupled with a LC-MS/MS assay provides fast and straightforward method development and greatly reduces the time for the tedious sample preparation procedures. In this work, a simple and sensitive bioanalytical method based on direct plasma injection using a single column high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS) was developed for direct cocktail analysis of double-pooled mouse plasma samples for the quantitative determination of small molecules. The overall goal was to improve the throughput of the rapid pharmacokinetic (PK) screening process for early drug discovery candidates. Each pooled plasma sample was diluted with working solution containing internal standard and then directly injected into a polymer-coated mixed-function column for sample clean-up, enrichment and chromatographic separation. The apparent on-column recovery of six drug candidates in mouse plasma samples was greater than 90%. The single HPLC column was linked to either an atmospheric pressure chemical ionization (APCI) or electrospray ionization (ESI) source as a part of MS/MS system. The total run cycle time using single column direct injection methods can be achieved within 4 min per sample. The analytical results obtained by the described direct injection methods were comparable with those obtained by semi-automated protein precipitation methods within +/- 15%. The advantages and challenges of using direct single column LC-MS/MS methods with two ionization sources in combination of sample pooling technique are discussed. PMID- 11885865 TI - Method for determination of histidine in tissues by isocratic high-performance liquid chromatography and its application to the measurement of histidinol dehydrogenase activity in six cattle organs. AB - A selective and simple HPLC procedure has been developed to determine histidine (His) and histidinol (HDL) in liver supernate. The separation was performed on a column, Mightysil RP-18 GP. The eluted analytes were measured with UV detection without derivatization which provided detection limits of 1.1 and 2.0 microM for His and HDL (S/N ratio, 3:1), respectively. Recovery of the analytes added to liver sample was 98.3-101.6% within a 1-day study and 95.7-98.6% on different day (6 days) studies. The apparent histidinol dehydrogenase activities (nmol/g wet tissue) at pH 8, 9, 10, 11, and 12 were 38.6, 50.4, 160.3, 274.3, and 185.6 for liver; 90.6, 132.2, 30.7, 22.1, and 6.76 for kidney; 0.0, 0.0, 38.2, 20.1, and 12.9 for pancreas; 0.0, 0.0, 0.0, 14.7, and 6.8 for spleen; 0.0, 0.0, 4.2, 6.8, and 0.0 for muscle; and 0.0, 0.0, 4.9, 1.8, and 0.0 for small intestine, respectively. On the basis of optimum pH values, histidinol dehydrogenase activity in the organs was in the following order: liver>kidney>pancreas>spleen>muscle>small intestine. PMID- 11885866 TI - The concept of nursing presence: state of the science. PMID- 11885867 TI - A non-theorist's perspective on nursing theory: issues of the 1990s. AB - The basis of all nursing endeavors, including practice and research, lies in theory. While nursing theorists are postulating and debating, practicing nurses are continuing with their daily routines and are often unaware that the world of nursing theory is changing. It is important, however, for all nurses to keep abreast of the latest developments in nursing theory. This article discusses some of the key developments within nursing theory based on a review of the nursing literature from 1990 through 1999. PMID- 11885868 TI - Conversation across paradigms: unitary-transformative and critical feminist perspectives. AB - The purpose of this article is to convey a conversation that occurred over a period of months between a unitary-transformative scholar and a critical feminist scholar. The intention of our conversation was to uncover, through dialogue and engagement, ways in which these two paradigms might help us understand the forces and conditions which impede and may liberate full expression of health and well being. Areas of essential tensions addressed were the relationships of action and theory, sense and soul, stories and numbers, and aesthetics and empirics. Critical conversational points were notions of liberation, consciousness and social conditions, unpredictability and acausality, and potentials for reconciliation that would serve nursing and society. We concluded that although there are significant differences that exist between the two paradigms, there are areas in which we might begin to speak with one voice for the betterment of nursing and health care. PMID- 11885869 TI - Positivism and qualitative nursing research. AB - Despite the hostility to positivism shown by qualitative methodologists in nursing, as in other disciplines, the epistemological and ontological instincts of qualitative researchers seem to coincide with those of the positivists, especially Bayesian positivists. This article suggests that positivists and qualitative researchers alike are pro-observation, proinduction, pro-plausibility and pro-subjectivity. They are also anti-cause, anti-realist, anti-explanation, anti-correspondence, anti-truth. In only one respect is there a significant difference between positivist and qualitative methodologists: most positivists have believed that, methodologically, the natural sciences and the social sciences are the same; most qualitative researchers are adamant that they are not. However, if positivism fails as a philosophy of the natural sciences (which it probably does), it might well succeed as a philosophy of the social sciences, just because there is a methodological watershed between the two. Reflex antagonism to positivism might therefore be a major obstacle to understanding the real reasons why qualitative research and the natural sciences are methodologically divergent; and less hostility on the part of qualitative nurse researchers might bring certain advantages in its wake. PMID- 11885870 TI - Intra-operative cardiac arrests. AB - This is a review of patients who had cardiac arrest in the operating suites at the University College Hospital (UCH), Ibadan between January 1994 and December 1998. The main objectives of the study were to evaluate the incidence of intraoperative cardiac arrests, identify associated factors and, in particular, those factors that might be essential for better prognosis. The study was retrospective, descriptive and cross-sectional. The figures were retrieved from the theatre records, the intensive care records, case notes and pathology reports. During the study period, 6,356 operations were performed. There were 35 cardiac arrests giving an incidence of 55 per 10,000 operations. Of the 35 cases, 18(51.4%) were males while 17(48.6%) were females. The age range was 4 months to 84 years (mean 32.5 years) with wide distribution through the decades. Of the twenty-four patients (68.6%) that were done as emergency cases, four patients (17.1%) recovered fully. Of the 11(31.4%) elective cases, 5 (41.7%) made full recovery. Ten patients (28.6%) were ASA I & II, while 25 (71.4%) were graded ASA status III to V. The factors associated with cardiac arrest in this study included emergency operation and the ASA status. PMID- 11885871 TI - A randomised trial to compare the efficacy and safety of Felodipine (Plendil) and Nifedipine (Adalat) retard in patients with mild-to-moderate hypertension. AB - The efficacy and tolerability of Felodipine extended-release was compared with Nifedipine retard in the management of patients with mild-to-moderate hypertension. A total of one hundred and thirty three patients were screened out of which one hundred and twenty-one patients were enrolled in a 9-week multicentre open, randomised rising-dose trial to receive either Felodipine 5-10 mg once daily or Nifedipine 10-20 mg twice daily. Blood pressure was measured at the end of the dosing interval that is 24 hours and 12 hours after Felodipine and Nifedipine respectively. Both drugs, Felodipine and Nifedipine were found to lower blood pressure significantly compared with baseline. After three weeks of treatment, seated blood pressure was reduced by 20/14 mmHg (systolic/diastolic) and by 24/16 mmHg after 6 weeks in the felodipine group. Corresponding values in the Nifedipine group were 16/09 mmHg and 24/13mmHg. Pulse rate was not significantly affected by either drugs. The percentage of patients who had satisfactory control after 3 weeks treatment was 57.6% for Felodipine and 33.3% for Nifedipine (significant). After dose titration (where necessary), at the end of the study the response rates were 76.3% (n=45) and 79.6% (n=43) for Felodipine and Nifedipine respectively (non significant). Both drugs were metabolically inert and did not derange the haematologic and biochemical profile of patients. They produced no significant weight changes. The pattern of side effects were similar in both groups but tended to be more severe with Nifedipine necessitating withdrawal of two patients in this group. In conclusion, Felodipine ER 5mg - 10mg once daily, and Nifedipine Retard, 20mg twice daily were equally effective medications for mild-to-moderate hypertension but Felodipine was better tolerated. PMID- 11885872 TI - High risk behaviours related to maternal and child health. AB - BACKGROUND: The excessive high maternal and perinatal mortality and morbidity figures characteristic of our community could be linked with maternal high-risk behaviours during pregnancy, delivery and postnatal periods. This cross-sectional questionnaire study assessed the prevalence of these behaviours among mothers in Ibadan. METHODS: Consecutive mothers attending infant welfare clinics of selected formal health facilities in Ibadan within 6 weeks of delivery were interviewed with a questionnaire. RESULTS: Of the 500 women interviewed, only 2.4% did not receive any antenatal care, 87.7% commenced care after first semester (57.2% and 30.5% in the second and third trimesters respectively). Twenty-five percent, 7.5% and 4.8% received no malaria prophylaxis, no tetanus immunisation, and iron preparations during pregnancy respectively. Seventeen percent (16.9%) received incomplete tetanus immunisation. Thirty percent (30%) of the women delivered out of formal health facilities, in church-based maternity and at home. While 91.4% received group breastfeeding counselling, 72% received group counselling specifically on exclusive breastfeeding. Eighty-four percent of the babies were not exclusively breast-fed, with 62.9%, 26.8% and 4.6% receiving supplementation with water, herbal preparations and artificial milk respectively. Sixty-seven percent of the babies received first immunisation after the first week of delivery. Prevalence of high risk behaviours related to maternal and child health care were not significantly different among the women utilising formal health facilities irrespective of the level, but were significantly higher among women who did not. CONCLUSION: A high prevalence of high-risk behaviours during pregnancy, delivery and postnatal periods, despite the availability of services, has been highlighted. There is a need to promote optimal utilization of existing services. There may be a need for quality assurance of these services. PMID- 11885873 TI - Observations on water supplementation in breastfed infants. AB - A total of 378 infants, under six completed months, were evaluated at the University of Benin Teaching Hospital, Benin City, Nigeria, to investigate some aspects of water supplementation in breastfed infants. Data were recorded concerning water supplementation and age at introduction of supplementary water among others. Water supplementation accounted for 91.7% of partial breastfeeding up to one completed month compared with 70-76% from up to 2 completed months to up to 5 completed months or earlier. The postnatal period of up to one month completed month may possibly be considered as the critical period for water supplementation since after this period, water was introduced only in a small proportion of infants during the first six months. It is suggested that strategies which prevent water supplementation at this critical period may ultimately reduce the prevalence of water supplementation during the first six months and, therefore, increase the rates and duration of exclusive breastfeeding in the studied population. PMID- 11885874 TI - Adult intussusception: the Jos experience. AB - Twenty two consecutive cases of adult intussusception managed between January 1990 and December 1998 at Jos University Teaching Hospital formed the basis of this study. Thirteen (59.1%) of the patients were males and 9(40.9%) females, with a male to female ratio of 1:4:1 and a mean age of 49.6 years. Most patients were referred late to our service as a result of poor index of suspicion and misdiagnosis. Laparotomy was done in all the cases and in 5(22.7%) patients no cause could be found, but in the remaining 17(77.3%) definite causes were identified which were mainly polyps in 7(31.8%) patients and colonic malignancies in 4(18%). The ileocolic intussusception was the commonest variety. Sixteen (72.7%) patients had bowel resection for colonic carcinoma, gangrenous bowel and irreducibility of the intussusception while manual reduction was successful in the other 6(27.3%) patients. The morbidity rate was 22.7% and the complications were wound infection and adhesive intestinal obstruction. Two deaths were recorded with a mortality rate of 9.1%. The pattern of adult intussusception as seen in the western world was observed in this tropical highland. PMID- 11885875 TI - Current views on epidemiology of renal tuberculosis. AB - The epidemiology of renal tuberculosis is characterised by a considerable variation in the frequency and distribution of the infection. It is difficult to be sure of the prevalence of renal tuberculosis (RTB throughout the world since survey results are to a certain extent dependent on the technique used. While the diagnosis of this disease is difficult, a high index of suspicion at all times with combined investigative tools permit the accurate diagnosis of most cases. Prevalence rates of over 5% in the general population have been reported in the Western countries. RTB is the commonest form of extrapulmonary tuberculosis among the Caucasians with male preponderance. There is paucity of data on the prevalence of RTB in tropical developing countries because of the general impression that this form of tuberculosis is rare in Blacks. However, hospital based retrospective data on extrapulmonary tuberculosis in Nigeria have revealed RTB prevalence rates of 1-3%. Tuberculosis involving the lymph nodes have been observed to be the commonest form of extrapulmonary tuberculosis in Nigeria. The age distribution of RTB obtained from different studies ranged from 14 to 70 years with the majority being below 50 years of age. Various male to female ratios ranging from 1:1 to 3:1 have been reported from different studies. Renal involvement in tuberculosis occurs through haemogeneous spread from a primary focus and is usually a bilateral disease. The most common primary site are the lungs. Many studies on pulmonary tuberculosis (PTB) outside Nigeria have shown that 26-75% of RTB co-exist with active PTB and 6-10% of screened open PTB patients have renal involvement. A recent study on RTB among active PTB patients in Ilorin, Nigeria revealed prevalence rate of 9.5% by urine Z-N positivity and 14% by utilising a combination of urine Z-N stain, sterile pyuria and renal tissue histology. It should be clear from the foregoing that RTB is not uncommon in both Caucasians and Blacks. It may be commoner in tropical developing countries than the Western World since PTB which is the main primary focus is endemic in these areas. PMID- 11885876 TI - An open study to evaluate the safety and efficacy of zafirlukast ("Accolate") in patients with mild to moderate asthma in Ibadan, Nigeria. AB - An open study to evaluate the safety and efficacy of Zafirlukast (oral leukotriene-receptor antagonist) in patients with mild to moderate asthma was conducted at the out-patient department of the University College Hospital (UCH), Ibadan. A total of 30 patients aged 16-70 years were screened into the trial but 20 patients that fulfilled the inclusion and exclusion criteria were enrolled in a 7-week open study to receive 20mg b.i.d. of "Accolate". Efficacy of treatment was assessed by changes in symptoms, beta-agonist use and pulmonary function tests (PEFR and FEV1). Safety was assessed by adverse experiences, laboratory test results, results of physical examination and chest-x-ray (when necessary). Zafirlukast (Accolate) significantly decreased daytime asthma symptoms scores (28.8%), night-time awakenings (80.76%), morning with asthma (36.4%) and beta agonist use (31.3%) and significantly increased the mean PEFR values (11.3%); FEV, (17.4%) at end point from their baseline values. Changes in symptoms, beta agonist use, and pulmonary function occurred within the first week of zafirlukast treatment and continued throughout the trial. Zafirlukast was well tolerated. Headache was reported in two patients. No significant changes were observed in laboratory test results, findings on physical examination. I concluded that zafirlukast produces early and sustained effects in the treatment of mild-to moderate asthma. PMID- 11885877 TI - Morbidity pattern in a sample of elderly Nigerians resident in Idikan community, Ibadan. AB - We documented the pattern of medical illnesses in 613 elderly Nigerians (398 females and 215 males) resident in Idikan community in Ibadan city. Their ages ranged from 65 to 110 years with a mean of 76.2 years. Medical disorders diagnosed either singly or in combinations were diagnosed in 364 (59.4%) subjects and there was no gender association. Cardiovascular problems were the commonest and high blood pressure (27.8%) was the most frequent diagnosis. Only 5 of the hypertensive subjects were aware of that diagnosis and were on regular medications. The complications presented with included heart failure and stroke. Visual impairment (12.1%) mainly due to cataracts and osteoarthritis (6.7%) in that order were next in frequency. The most frequent neurological disorders were hearing impairment and movement disorders. The other conditions encountered were similar to the findings in previous studies in this environment, and the usual findings in studies focusing on this age-group in other countries. The presence of morbidity was significantly associated with increasing age and poor performance on screening. The latter increased the probability of being selected for clinical examination with detection of medical problems or could suggest associated cognitive impairment. The prevalence of systemic hypertension was not different from findings in other communities in people of similar age groups. This study emphasises the role of hypertension as a major cause of morbidity in this community and stresses the need for increased health awareness especially with regards to regular checking of blood pressure so as to avoid complications. PMID- 11885878 TI - Human immunodeficiency seropositivity among mother-child pairs in South West Nigeria: a community-based survey. AB - A community based survey to determine the prevalence of human immunodeficiency infection in Nigerian women and children in South Western Nigeria is reported. A multi-stage cluster random sampling procedure was used to select mother-child pairs from 35 enumeration areas in South western Nigeria. The final study sample consisted of 460 mothers and 476 children (including 16 sets of twins). A commercially available recombinant antigen-based ELISA method was used to test for HIV-1 and HIV-2 antibody in sera and Western blotting was used as a confirmatory test for initially reactive samples. Only one mother-child pair (out of 460 mother-child pairs) was found to be positive for HIV antibody giving a mother-child concordance for HIV infection of 0.22%. Antibody to either HIV-1 or HIV-2 was detected in 3.8% (18/476) of the children's sera and in 43% (20/460) of mothers sera. HIV-1 reactivity was commoner than HIV-2 reactivity (2.9% versus 0.8% among children and 2.8% versus 1.5% among mothers). There were many more positive samples in the rural than in urban areas among children (7.1% versus 1.1%) and also among mothers (6.8% versus 2.4%), (p<0.001). Thus, HIV infection appears to be a real problem in South western Nigeria. The lack of concordance between mother-child sera suggests that vertical transmission may not be a major route of transmission of HIV infection in children in South western Nigeria. It is suggested that certain high risk practices (such as the re-use of unsterilised hypodermic needles for injections and surgical knives in local scarification) which are common practices, especially in rural areas, need to be investigated as potential major modes of transmission of the infection. Control programmes need to take note of these findings in order to adequately plan comprehensive health education which will cover the whole population, including children. PMID- 11885879 TI - Time of presentation for treatment and profile of deformities among leprosy patients in South Eastern Nigeria. AB - A 10 year review of leprosy patients seen at Leprosy Hospital Ekpene Obom in South Eastern Nigeria (1988-1997) was carried out to evaluate the effect of early identification and treatment of leprosy patients in the limitation of deformities among them. A total of 2,597 patients comprising 1,714 (66%) males and 883 (34.0%) females formed subjects for the study. Of these 288(11.1%) were aged 15 and below while 2,309 (88.9) were above 15 years. Their case records were thoroughly reviewed noting the duration of disease before presentation, type and location of deformity as well as the type of leprosy. Though there was a steady decline in the total number of leprosy patients seen over the study period as well as a decrease in the mean duration of illness before presentation, approximately 19% of patients still had deformities at presentation, a figure much lower than those reported by other workers. Analysis of the pattern of deformities shows that most patients 71.2% presented with affectation of the upper and lower limbs with consequent functional disability. We conclude that early treatment is an effective means of reducing the prevalence of deformity and thus disability from leprosy. More effective implementation of health education and treatment programmes initiated by the W.H.O should further reduce the scourge of leprosy in our community. PMID- 11885880 TI - Hirschsprung's disease in children in South Eastern Nigeria. AB - In a three year prospective study between January 1991 and December 1993 on 21 children with Hirschsprung's disease in our environment, the main complaint were abdominal distention, constipation, wasting, diarrhoea and retardation in growth in decreasing order of frequency. As in other reports a male preponderance of approximately 4:1 was observed in the study. Patients presented late to hospital and mostly when the complications of complete intestinal obstruction are obvious. This is normally when the habitual enema can no longer afford anymore relieve. A palliative transverse colostomy was considered essential in our environment whereby the complications normally associated with outright resection and anastomosis can be minimised. Public health education as to seeking early medical assistant is necessary. This may prevent fatal complications as some of the childhood mortality in our environment can be attributed to ignorance on the parts of parents. PMID- 11885881 TI - Factors affecting angular deformities of the knees in Nigerian children--Ilorin experience. AB - 44 children with angular knee deformities were studied at the University of Ilorin Teaching Hospital from September 1994 to December, 1997. There were 19 males (43.2%) and 25 females (56.8%). The commonest age range was 0-5 years age range (81.5%). Bilateral angular deformities are more common than unilateral angular deformities. Windswept deformities accounted for 16 cases(36.4%), bilateral Genu Varum accounted for 15 cases (34.0%), and unilateral Genu Valgum, 8 cases (18.2%). It was found in the study that 40 cases (90.9%) were from urban communities while 4 cases (9.1%) were from rural communities. The angular knee deformities affect both Christian and Moslem children alike; however, the environmental factor is the restriction of children indoors which is a serious factor in the causation of Nutritional Rickets. Angular deformities of the knee were far more common amongst urban dwellers than in children from rural area (Ratio 10:1) while involvement of both knees (88.6%) simultaneously is also more common than single knee involvement (11.4%). Some of the objectives of the investigation were to find out the implications of the angular deformities to parents and children; and ways to prevent occurrence were also discussed in this paper. PMID- 11885882 TI - Malignant colorectal tumours: a ten year review in Jos, Nigeria. AB - One hundred and forty-four cases of histologically confirmed colorectal cancer in patients managed at the Jos University Teaching Hospital (JUTH) over a 10 year period from January, 1989 to January, 1999 is discussed with special consideration to incidence, distribution and unfavourable prognosis. Altogether, 144 patients were treated for colorectal carcinoma. Eighty-seven were males while fifty-seven were females, giving a male to female ratio of 1.51:1. The mean age was 44.3 years. The commonest clinical presenting features were weight loss, bloody mucoid diarrhoea, anorectal mass, anaemia, low-back pain and constipation/increased noise in the abdomen, present for not less than 3 months. The rectum and rectosigmoid junction were the commonly affected sites. All, except four patients, had advanced disease at first presentation. Treatment was basically palliative with only 43.5 percent of those offered such treatment alive at 6 months while 25 percent had died. Prognosis is unfavourable. Though, predisposing factors are not clear, promotion of educational programme highlighting the dangers of concealing chronic large bowel symptoms and screening efforts will most probably reduce morbidity and mortality rates associated with this condition. PMID- 11885883 TI - Factory floor injury in a Lagos sawmill. AB - We retrospectively reviewed injuries sustained in 36 consecutive accidents in a wood-processing factory and managed at a private hospital over a 2-year period in Lagos. The commonest injuries were lacerations by revolving saws, followed by crush injuries from entrapment by machines and from falling logs and planks. The upper limbs were involved in 24 (66%) of these accidents cases. Of 137 workers on the factory floor, the highest injury rate (64%) occurred among machine operators. While 80.6% of these injuries were simple ones treated by suturing and dressing, 7 (19.4%) were life-threatening enough to warrant hospitalisation and major surgery, with 6 sustaining a mean permanent disability of 7.1 +/- 6%. Although factory-floor injuries constituted only 6.5% of 553 hospital attendance recorded within the period from the company, they were responsible for 44.2% of total medical expenditure by the company within the same period. Non-use of protective gears and disregard for safety procedures were noted in most of the accidents. The in-house first-aid program was adjudged as life-saving in the few major cases managed. We concluded that while many factory-floor injuries in wood processing factories may be minor hand injuries, provision and strict observance of safety protocols as well as an active first-aid program are invaluable to minimise morbidity, cost and loss of productive man-hours in wood processing factories. PMID- 11885884 TI - Cardiovascular involvement in HIV/AIDS: report of 3 cases. AB - Three cases of dilated cardiomyopathy in patients with IIIV/AIDS are being reported. The three patients are of young age group and they presented with cardiac symptoms for the first time. They were all heterosexuals and not known was as intravenous drug abuser. There was no history of rheumatic fever or hypertension or diabetes mellitus and ischaemic heart disease. Examination confirmed cardiac failure and investigations including chest x-ray, echocardiograph and electrocardiograph confirmed dilated heart. All the patients tested positive to HIV-1 antibodies. They were managed with the usual anticardiac failure regimen. Two of the patients died on admission, one developed multi organ failure and the other had tonic-clonic seizure. In other parts of Africa reports have also emerged describing the cardiovascular involvement in HIV/AIDS. PMID- 11885885 TI - Case report--abdominal cocoon. AB - Abdominal cocoon is a rare cause of acute intestinal obstruction seen almost exclusively in young adolescent females. Almost all cases are diagnosed at surgery and cured by excising the fibrous cocoon. This case although diagnosed accidentally too was treated conservatively successfully. PMID- 11885886 TI - Fascia lata autograft for orbital floor reconstruction. AB - Enophthalmos and diplopia are two major complications that result from a downward displacement of the eyeball. A case in which a fascia lata autograft was used to prevent these complications is presented. When covered by a flap with rich blood supply, this autograft can be used as a good substitute for alloplastic materials. PMID- 11885887 TI - Acute haemorrhagic pancreatitis in HIV positive patients. AB - We present two cases of severe acute haemorrhagic pancreatitis (1,2), in otherwise healthy adults who were HIV positive. Acute pancreatitis is not known to be common in the African communities but the incidence is on the increase (1). Both of them scored between 5 and 6 points on the Ransom scale (3). One of them died despite similar aggressive resuscitation, adequate transfusion with fresh frozen plasma (4,5) and peritoneal lavage (6,7,8) Though Steinberg and Tenner (2) had shown a higher incidence (4-22%) of acute pancreatitis among patients with the acquired immune deficiency syndrome (AIDS) in some populations, we are not aware of any observation in literature that same is true in otherwise healthy patients who are HIV positive. We are posting that what we have observed may indeed be human immune deficiency viral haemorrhagic pancreatitis. A prospective study of patients with acute pancreatitis will determine the position. PMID- 11885888 TI - Influence of storage days on the distribution for time of embryonic mortality during incubation. AB - Breakout analysis of 11,254 chicken eggs that failed to hatch was used to assess the influence of storage days on the distribution for time of embryonic mortality during incubation and on reproductive efficiency. Eggs were collected over 30 d, stored from 2 through 18 d, and incubated in two hatches. For each storage day within hatch, proportions of embryonic mortality during each of the 21 d of incubation, among embryos that did not survive incubation, were fitted by a diphasic Weibull distribution. Multivariate analysis was used to assess the influence of hatch and storage days within hatch on parameters of the distribution and on two measures of reproductive efficiency, proportions of embryonic mortality during incubation among all eggs incubated P(mort) and among fertile eggs incubated P(mort/fert), and to obtain partial correlation coefficients. Storage days influenced the distribution for time of embryonic mortality in each hatch, but the effect was different for each hatch. As the number of storage days increased, P(mort) and P(mort/fert) increased. Partial correlations showed that P(mort) and P(mort/fert) decreased as the proportion of embryos that died during the first phase decreased and as duration of the second phase increased. The shape of the distribution for time of mortality during incubation influenced reproductive efficiency. Factors that influence the shape of this distribution, other than hatch and storage days within hatch, should be studied to increase reproductive efficiency in the poultry industry. PMID- 11885889 TI - Phenotypic traits as reliable indicators of fertility in male broiler breeders. AB - Genetic selection procedures applied to improve broiler performance may negatively impact the subsequent reproductive efficiency of breeders, particularly in males. Identification of traits that reliably indicate individual male fertility would facilitate selection for reproduction. We hypothesized that physical traits, such as comb area, relative testicular weight, and testicular weight asymmetry, may correlate with fertility in two male-selected primary broiler breeder strains (A and B). Thirty males per strain, individually housed with an average of 10 females, were evaluated at five age periods within the 30 to-50-wk breeding cycle. Flock fertility by candling eggs at Day 19 of incubation and sample fertility by visual assessment of the germinal disc were determined. Sperm penetration (SP) through the perivitelline layer was assessed. Comb area was evaluated by image analysis at 40 and 50 wk, and relative testicular weight was measured at 50 wk. Strain A sample and flock fertility (P < 0.001) and SP values (P < 0.0001) were significantly lower than Strain B. Both strains had a significant decline of fertility and SP with age (P < 0.0001). Strain A comb area correlated with sample fertility (P < 0.05), flock fertility (P < 0.05), and relative testicular weight (P < 0.01). Conversely, Strain B relative testicular weight correlated with sample fertility (P < 0.0001) and flock fertility (P < 0.001). Significant correlations were not found between testicular weight asymmetry and other reproductive traits. Results suggest comb area may be a reliable indicator of male fertility in Strain A. PMID- 11885891 TI - Tenderization of hot-boned broiler breast meat by clamping during chilling. AB - Hot-boned broiler breast fillets were tightly clamped between rigid aluminum plates during chilling to determine whether tenderness is increased if breast fillets are not allowed to shorten during rigor. In two experiments, 6-wk-old broilers were processed in a pilot plant. Approximately 5 min after evisceration, the breast fillets (pectoralis major) were deboned, and each fillet was subjected to one of two treatments while chilling for 2 h in ice slush. Fillets were placed in perforated plastic bags (hot-boned control) or clamped between rigid aluminum plates that compressed the meat to a uniform thickness of 7.2 mm during chilling. In Experiment 2, chilling time in ice slush was 1 h, and a third treatment was added to make an incomplete block design in which one breast half was left intact on the carcass and was deboned immediately after chilling. All breast fillets were sealed in plastic bags after the chilling period, held overnight at 4 C, and then cooked at 85 C for 30 min in a steam kettle. In Experiment 1, clamping for 2 h reduced Warner-Bratzler shear values of hot-boned fillets from 11.4 to 2.7 kg. In Experiment 2, shear values for the treatments were 13.0, 9.2, and 5.1 kg for the hot-boned, cold-boned, and hot-boned clamped treatments, respectively, with significantly lower shear values for the clamped fillets. Clamped fillets were significantly thinner than the control fillets in both experiments. Cooked yield as a percentage of postchill weight was significantly higher for the clamped compared to the hot-boned control pieces, 81.1 versus 77.3%, with cold-boned pieces being intermediate and not different from the other treatments. Shear values were reduced, and cooked yield was increased by clamping hot-boned fillets during chilling. PMID- 11885890 TI - Identification of ovotransferrin as an acute phase protein in chickens. AB - Inflammation is homeostatic process associated with a variety of cellular injuries resulting from infections, toxicosis, and physical trauma. The studies on inflammation in avian species are limited. To understand the inflammation induced changes, 4-wk-old male broiler chickens were subjected to experimental inflammation by a subcutaneous injection of croton oil (inflammatory) with changes in serum measured over time and were compared with birds treated similarly with olive oil (injected control). Croton oil treatment significantly elevated serum interleukin (IL)-6 concentrations and heterophil counts by 6 and 16 h postinjection, respectively, which returned to the basal levels of controls at 16 and 24 h, respectively. Croton oil treatment affected the serum protein profiles of chickens as assessed by SDS-PAGE and densitometric analyses. Compared with olive oil-injected or noninjected chicken sera, there were increases in the density of protein bands corresponding to molecular weights (MW) of 42, 65, 200, and 219 kDa and decreases in bands corresponding to 49 kDa (serum albumin) and a 56-kDa protein in chickens treated with croton oil. Most of these changes were evident at 24 h and lasted through 48 h. The protein band corresponding to 65 kDa was further characterized using two-dimensional gel electrophoresis and N terminal sequence analyses. A sequence similarity search in the Genbank database using the first 22 amino acids yielded a complete homology with chicken ovotransferrin. Western blot analysis using antichicken serum transferrin or antichicken ovotransferrin antibodies also confirmed the 65-kDa protein band to be ovotransferrin. Under nonreducing conditions, the ovotransferrin standard also showed an apparent MW corresponding to 65 kDa, like the serum transferrin. The serum ovotransferrin was found to be glycosylated using a glycoprotein stain. Although the significance of ovotransferrin in avian inflammation is not clear, these results show that it is a major acute phase protein (APP) in chickens. PMID- 11885892 TI - Variation in numbers of bacteria on paired chicken carcass halves. AB - Bacterial counts from paired broiler carcass halves were examined for relationships between numbers and kinds of bacteria that might indicate fecal contamination. Broiler carcasses removed from a commercial processing plant just before chilling were split aseptically along the midline, and each side was rinsed in 400 mL of phosphate buffered saline for 1 min with either mechanical or hand shaking. Both halves of six carcasses were rinsed on four different days for a total of 24 carcasses sampled with each shaking method. Aerobic bacteria, coliforms, Escherichia coli, and Campylobacter jejuni were enumerated and summed to obtain whole carcass counts. There were no significant (P < 0.05) differences in numbers of bacteria recovered by the two rinse methods. In left-right comparisons, only E. coli was significantly different (P = 0.04), with the right side having higher counts (least-square means of 1.09 vs. 0.97). For aerobic plate count (APC), coliforms, E. coli, and Campylobacter, correlations between paired left and right side counts were between 0.78 and 0.86. The correlation between whole carcass counts and absolute left-right differences was significant for APC (0.43), but was not significant for coliforms, E. coli, and Campylobacter, so higher whole carcass counts were not associated with higher counts on one side of the carcass. Correlations between different bacteria on whole carcasses were significant for E. coli-APC (0.39), E. coli-coliforms (0.67), and APC-coliforms (0.71), but other combinations had non-significant correlations. The correlation was not significant between E. coli and Campylobacter, a relatively fragile organism whose presence can be interpreted to indicate fairly recent fecal contamination. There were no indications that high E. coli counts on inspection-passed, prechill carcasses indicated recent fecal contamination. PMID- 11885893 TI - Microbiological consequences of skin removal prior to evisceration of broiler carcasses. AB - The objective of this project was to determine if removal of skin prior to evisceration lowers the number of bacteria that can be recovered by whole carcass rinse or sponge sampling. Four experiments were conducted, two with each type of sampling (rinse or sponge). New York dressed carcasses obtained from a commercial broiler processing plant were aseptically skinned or left with skin intact. The carcasses were then aseptically eviscerated by hand. Carcasses were rinsed in 100 mL sterile water or sampled by moist sponge. When sampled by rinse, significantly fewer Campylobacter and total aerobic bacteria were recovered from carcasses that had been skinned prior to evisceration. When sampled by sponge, significantly fewer Campylobacter, Escherichia coli, coliform and total aerobic bacteria were recovered from the outer surface of carcasses without skin. No differences were noted for bacterial counts recovered from internal surfaces by sponge sampling. Similar trends were observed when carcasses were subjected to an inside and outside washing step after evisceration. Removal of skin and washing the carcass led to significantly less Campylobacter being recovered by whole carcass rinse compared to carcasses that were washed with the skin on. When sampled by sponge, incidence of Campylobacter and level of total aerobic bacterial counts were lower on the outer surface of skinned and washed carcasses than on washed carcasses with intact skin. Like the unwashed carcasses, no differences were noted for bacterial counts recovered from internal surfaces by sponge sampling. Although not commercially practical, it is possible to lower the level of Campylobacter on the outside of broiler carcasses by removal of the skin prior to evisceration. PMID- 11885894 TI - Effect of physical feed restriction during rearing on large white turkey breeder hens: 2. Reproductive performance. AB - Large White turkey breeder hens were used to evaluate the effect of three different levels of physical feed restriction on subsequent reproductive performance. The feed treatments were: 1) fed ad libitum throughout the study (CC), 2) feed-restricted from 16 to 24 wk (CR), 3) feed-restricted from 3 to 16 wk (RC), and 4) feed-restricted from 3 to 24 wk (RR). Feed restriction was implemented so that restricted-fed hens (RC and RR) achieved a 45% reduction in BW as compared to CC hens at 16 wk. From 16 to 24 wk, feed was allotted to RR and CR hens to maintain a slight increase in BW. At the completion of the respective restriction periods, hens were gradually returned to ad libitum feeding. At 30 wk of age, hens were photostimulated for a 20-wk summer season egg production cycle. Hens receiving RC and RR treatments laid significantly more eggs than did CC and CR hens for the first 5 wk of lay. However, once the house temperature increased to 26.7 to 29.4 C during 6 to 10 wk of lay, egg production of all hens decreased, resulting in a significant decrease in cumulative egg production for RR and RC hens compared to CC and CR hens. Egg and poult weights were less for RC and RR hens compared to those from CC and CR hens. In conclusion, age of breeder, season of implementation, and length of physical feed restriction have significant effects on the reproductive performance of turkey breeder hens. PMID- 11885895 TI - Market age live weight, carcass yield, and liver characteristics of broiler offspring from breeder hens fed diets differing in fat and energy contents. AB - The effects of energy level, fat type, and fat level in breeder hen diets on subsequent offspring market age live BW, carcass yield, and liver characteristics from breeder hens at 29 and 36 wk of age were evaluated. At 22 wk of age, six dietary treatments were imposed. Dietary treatments contained: 1) 3.0%, added poultry fat (PF) and 467 (high energy) kcal/hen per day at peak production (CPP), 2) no added fat and high energy, 3) 3.0% added PF and 430 (low energy) CPP, 4) no added fat and low energy, 5) 1.5% added PF and 449 (moderate energy) CPP, and 6) 3.0% added corn oil (CO) and moderate energy. Breeder age influenced Day 43 broiler live BW, percentage total carcass and front-half yields, and liver moisture contents. Furthermore, wet and dry liver weights were higher in female broilers compared to those of male broilers from 29-wk-old breeder hens. Live BW was higher in broilers from hens fed low-energy diets compared with moderate energy diets and 3.0% compared to 1.5% PF diets. Percentage liver DM was higher in females compared to male broilers from hens fed 3.0% CO and moderate energy and was highest in male and female broilers from hens fed 1.5% PF and moderate energy. Percentage wet liver weight and liver DM were higher and liver moisture content was lower in broilers from hens fed 1.5% compared to 3.0% PF diets. Overall, energy and fat levels in breeder diets had subsequent influences on market age weight and liver characteristics of broilers. PMID- 11885896 TI - Fatty acid composition and egg components of specialty eggs. AB - Egg components, total fat, and fatty acid content of specialty eggs were compared. One dozen eggs were collected and analyzed from each of five different brands from hens fed a diet free of animal fat (SP1), certified organic free range brown eggs (SP2), uncaged unmedicated brown eggs (SP3), cage-free vegetarian diet brown eggs (SP4), or naturally nested uncaged (SP5). Regular white-shelled eggs were the control. A significant (P < 0.05) difference was observed in the egg components and fatty acid content in different brands. The percentage of yolk was lower (P < 0.05) in SP2 and SP4 with a concomitant increase (P < 0.05) in the percentage of white. The percentage of shell was lower (P < 0.05) in SP4 and SP5. The total edible portion was greater in SP4 and SP5. The yolk:white ratio was greater (P < 0.05) in SP3. The total lipid content was lower in SP4 eggs. The content of palmitic (C16:0), stearic (C18:0), and total saturated fatty acids were lower (P < 0.05) in SP1. No difference was observed in the content of palmitoleic (C16:1), oleic (C18:1), or total monounsaturated fatty acids. The content of n-3 fatty acids in SP2, SP4, and SP5 were similar to control eggs. The ratio of total n-6:n-3 polyunsaturated fatty acids ranged from 39.2 for SP5 to 11.5 for SP1 (P < 0.05). No difference was observed in the total polyunsaturated fatty acid content of eggs (P > 0.05). PMID- 11885897 TI - Prevention of Salmonella enteritidis infection in commercial ducklings by oral chicken egg-derived antibody alone or in combination with probiotics. AB - Ducklings were given egg-derived antibody against Salmonella enteritidis (Ab) in drinking water daily to determine if infection could be prevented. Pekin ducklings in all experimental groups were infected on Day 1 or 5 with 0.7 x 10(6) Salmonella enteritidis (SE). Spleen, liver, and intestine of each bird were collected and cultured on Days 7, 14, 21, and 28. Only livers and spleens were culture positive for SE. Ducklings infected on Day 1 had more SE infections than controls at each observation. Ducklings infected on Day 5 had fewer SE infections than controls on Days 7, 14, and 21. The same experiment was repeated to determine if SE infection could be prevented under production conditions. Only 10 ducks per group were infected with 1.02 x 10(7) SE. In addition to Ab, one group each, infected on Day 1 or 5, received a proprietary probiotic (Pro) daily to determine if Pro was synergistic to Ab. Groups receiving Ab and Pro and infected on Day 1 had fewer birds infected than Ab alone in Day 1-infected birds. Both Day 1-infected groups had more birds infected than controls. Birds infected on Day 5 had fewer ducks infected than controls on Days 7, 14, and 21. Except for Day 14, birds receiving both Ab and Pro and infected on Day 5 had fewer birds infected than Ab alone on Day 21 and 28. Probiotics act synergistically with oral Ab. Oral antibodies may serve as a tool to prevent salmonella infection in poultry. PMID- 11885898 TI - Interaction of dietary vitamin E with Eimeria maxima infections in chickens. AB - In two trials, broiler chickens, processed similarly to those placed in commercial operation, were fed, from 1 d of age, a range (13 to 200 ppm) of DL alpha-tocopheryl acetate (VE-AC) levels, and the effects on the pathology of Eimeria maxima infections were assessed at 6 d postinoculation (PI). In Trial 1, dietary levels of VE-AC had little significant effect on variables characterizing pathology except for the number of oocysts shed, which was significantly increased in chicks treated with higher VE-AC levels. The infection was judged to be mild based on moderate lesion scores (2.2+/-0.2), lack of significant effects on weight gain (7+/-1.6% decrease), moderate reduction in plasma carotenoids (21+/-2%) and small increases in plasma NO2-+NO3- (141+/-12%). In uninfected and infected chickens, plasma alpha-tocopherol (AT) increased with dietary levels of VE-AC; however, E. maxima infection caused a fairly constant decrease in AT of 35.3+/-3.2% across these levels. Plasma gamma-tocopherol (GT) levels were unaffected by dietary VE-AC or E. maxima infection. In Trial 2, pathology, again, was relatively unaffected by dietary VE-AC level. The infection was judged to be severe based on lesion scores (3.5+/-0.1), reduction in weight gain (30.7+/-3%), plasma carotenoids (72.4+/-1.5%), uric acid (16.3+/-3.4), albumin (37.8+/-2.8%), large increases (261+/-8%) in plasma NO2-+NO3-, and high numbers of oocysts shed per chick (4.12+/-0.4 x 10(7)). Plasma AT again increased with increasing dietary VE-AC levels in uninfected and infected chicks, but the mean decrease across VE AC levels caused by E. maxima infection was 73.14+/-3.3%. GT levels were erratic and unrelated to dietary VEAC or infection. Thus, in processed broiler chickens, high dietary VE-AC did not prevent or lessen the pathology caused by mild or severe infections with E. maxima. The main effect of E. maxima infection appeared to be reduction in plasma AT levels. We postulate that this reduction may be due to malabsorption of AT, which results from physical damage to the absorptive mucosa, reduction in esterases required to hydrolyze the VE-AC, and a generalized lipid malabsorption, preventing movement of the free AT to circulating blood and infected tissues. PMID- 11885899 TI - Reducing airborne pathogens, dust and Salmonella transmission in experimental hatching cabinets using an electrostatic space charge system. AB - Electrostatic charging of particles in enclosed spaces has been shown to be an effective means of reducing airborne dust. Dust generated during the hatching process has been strongly implicated in Salmonella transmission, which complicates the cleaning and disinfecting processes for hatchers. Following two preliminary trials in which dust reduction was measured, four trials were conducted to evaluate the effectiveness of an electrostatic space charge system (ESCS) on the levels of total aerobic bacteria (TPC), enterobacteriaceae (ENT), and Salmonella within an experimental hatching cabinet. The ESCS was placed in a hatching cabinet that was approximately 50% full of 18-d-old broiler hatching eggs. The ESCS operated continuously to generate a strong negative electrostatic charge throughout the cabinet through hatching, and dust was collected in grounded trays containing water and a degreaser. An adjacent hatching cabinet served as an untreated control. Air samples from hatchers were collected daily, and sample chicks from each hatcher were grown out to 7 d of age for cecal analysis in three of the trials. The ESCS significantly (P < 0.05) reduced TPC and ENT by 85 to 93%. Dust concentration was significantly reduced (P < 0.0001) during the preliminary trials with an average reduction of 93.6%. The number of Salmonella per gram of cecal contents in birds grown to 7 d of age was significantly (P < 0.001) reduced by an average log10 3.4 cfu/g. This ionization technology is relatively inexpensive and could be used to reduce airborne bacteria and dust within the hatching cabinet. PMID- 11885900 TI - Chronic effects of fumonisin B1 in broilers and turkeys fed dietary treatments to market age. AB - Floor pen studies were conducted with 270 broiler chicks and 144 turkey poults, all 1 wk old, to evaluate the chronic effects of fumonisin B1 (FB1). A completely randomized design was used in both studies with six pen replicates of 15 chicks or eight pen replicates of six poults assigned to each of three dietary treatments from Weeks 1 to 7 (broilers) or to Week 14 (turkeys). Fusarium moniliforme (M-1325) culture material was added to a typical corn-soybean basal diet to supply 0, 25, or 50 mg FB1/kg diet. Feed intake, body weight gain, and feed conversion of chicks were not affected (P > 0.05) by FB1. Turkeys fed 50 mg FB1/kg had significantly (P < 0.05) lower feed intake than the controls. Compared with controls, chicks and turkeys fed FB1 diets had significantly higher liver sphinganine to sphingosine ratios (P < 0.05). Relative organ weights of chicks were not affected (P > 0.05) by FB1, other than those chicks fed 25 mg FB1/kg, which had lower (P < 0.05) relative proventriculus weights than the chicks fed 0 or 50 mg FB1/kg. Broilers fed 50 mg FB1/kg had decreased serum calcium and increased serum chloride when compared to broilers fed 0 or 25 mg FB1/kg. Hematology was not affected (P > 0.05) by dietary FB1. No lesions were present in any organ examined microscopically. Results indicate that 50 mg FB1/kg diet is detrimental to turkeys but is not toxic to broilers fed to market age. PMID- 11885901 TI - Use of cool perches by broiler chickens. AB - Broilers under commercial conditions might experience relatively high temperatures during summer and leg disorders year round that may be partially alleviated by providing them with access to cooled perches. It is unknown, however, how perch temperature and factors such as height and position of the perch affect perch use. Furthermore, little is known regarding gender effects. Eight thousand 1-d-old, mixed-sex broilers were exposed to three perch treatments to determine preferences for water-cooled perches over ambient temperature perches and preferences for height, location, and temperature section of the perch. The experimental treatments were as follows: 1) three cool perches 15 cm above the floor (Cool 15), 2) three ambient perches 7.5 cm off the floor (Ambient 7.5), 3) three ambient perches 15 cm high (Ambient 15), and 4) control chambers with no perches. Total number of birds perching, their positions, and temperature section within the perch were recorded. The results indicate a strong preference for high perches as birds grow (P < 0.0001). The cooler sections of the perch were utilized more than warmer sections within the cool treatments (P < 0.05). Females showed a stronger tendency to perch than males, particularly within the cool treatment (P < 0.0001). The higher perch use could be one of the reasons for the higher eviscerated body weight found in females with access to cool perches (P < 0.05). Differences in mean body weight were not significant (P = 0.07). Potential beneficial effects of perch access in final body weights needs to be further investigated. PMID- 11885902 TI - Fluorescent marker for the detection of crop and upper gastrointestinal leakage in poultry processing plants. AB - Previous published research has identified the crop as a source of Salmonella and Campylobacter contamination for broiler carcasses and reported that broiler crops are 86 times more likely to rupture than ceca during commercial processing. Presently, we evaluated leakage of crop and upper gastrointestinal contents from broilers using a fluorescent marker at commercial processing plants. Broilers were orally gavaged with a fluorescent marker paste (corn meal-fluorescein dye agar) within 30 min of live hang. Carcasses were collected at several points during processing and were examined for upper gastrointestinal leakage using long wavelength black light. This survey indicated that 67% of the total broiler carcasses were positive for the marker at the rehang station following head and shank removal. Crops were mechanically removed from 61% of the carcasses prior to the cropper, and visual online examination indicated leakage of crop contents following crop removal by the pack puller. Examination of the carcasses prior to the cropper detected the marker in the following regions: neck (50.5% positive), thoracic inlet (69.7% positive), thoracic cavity (35.4% positive), and abdominal cavity (34.3% positive). Immediately prior to chill immersion, 53.2% of the carcasses contained some degree of visually identifiable marker contamination, as follows: neck (41.5% positive), thoracic inlet (45.2% positive), thoracic cavity (26.2% positive), and abdominal cavity (30.2% positive). These results suggest that this fluorescent marker technique may serve as a useful tool for rapid identification of potential changes, which could reduce the incidence of crop rupture and contamination of carcasses at processing. PMID- 11885903 TI - Influence of dietary phosphorus on performance of Hy-line W36 hens. AB - Two experiments were conducted to determine the effect of nonphytate phosphorus (NPP) on hen performance in an environmentally controlled house. In Experiment 1, 21-wk-old Hy-Line W36 hens (n = 1,248) were randomly assigned to 13 dietary treatments (0.1 to 0.7% NPP, at graded increments of 0.05%) for 17 wk. In Experiment 2, 45-wk-old Hy-Line W36 hens (n = 960) were randomly allocated to eight diets in a 2 x 4 factorial arrangement of treatments. Two levels of Ca (3 and 4%) and four levels of NPP (0.1, 0.2, 0.3, and 0.4%) were fed for 8 wk. Feed consumption (FC), egg production (EP), egg weight (EW), egg specific gravity (ESG), bone density (BD), bone mineral content (BMC), bone breaking strength (BBS), BW and mortality were evaluated to determine performance. Results of Experiment 1 indicated that FC, EP, and BW increased as NPP was increased from 0.1 to 0.7% (77 to 588 mg/hen/d). During Week 12, a sharp decrease in FC and EP was observed in hens fed the most deficient level of 0.1% NPP. Egg weight and ESG decreased linearly (P < 0.05) as dietary NPP was increased from 0.1 to 0.7%. However, after 14 wk, ESG decreased (P < 0.05) in hens fed 0.1% NPP. Bone breaking strength was higher (P < 0.05) in hens fed 0.3 to 0.4% NPP, indicating maximum bone quality. Mortality was higher (P < 0.05) in hens fed 0.01% NPP, followed by hens fed 0.15% NPP. In Experiment 2, a pronounced adverse effect of P deficiency was observed on FC and EP within 2 wk compared with 12 wk in Experiment 1. Reduction of NPP to 0.1% reduced BD, BMC, BBS and increased hen mortality (P < 0.05). Reducing dietary Ca from 4 to 3% decreased ESG (P < 0.05). A wide variation in response time to P deficiency indicated that P requirement varied for different performance criteria with age. PMID- 11885904 TI - Dietary interaction of 1,4-diaminobutane (putrescine) and calcium on eggshell quality and performance in laying hens. AB - An experiment was conducted to evaluate the potential dietary interaction between 1,4-diaminobutane (putrescine) and calcium on eggshell quality and overall laying performance. One hundred ninety-two 30-wk-old White Leghorn hens were fed a corn and soybean-meal-based diet supplemented with 0.00, 0.05, 0.10, or 0.15% putrescine and 2.5, 3.0, 3.5, or 4.0% calcium in a factorial design (12 birds per diet) for 4 wk. The percentage of egg production increased linearly (P < 0.05) with increasing levels of dietary calcium. Significant interactions (P < 0.05) were observed between dietary putrescine and calcium for eggshell thickness, eggshell deformation, percentage of eggshell, calcium intake, total calcium retention, total eggshell calcium, and percentage of eggshell calcium. Interactions were due to quadratic effects of putrescine or calcium on these parameters. Eggshell thickness and percent eggshell increased when hens were fed 3.5% calcium in combination with 0.1% putrescine; however, calcium intake and calcium retention were significantly lower (P < 0.05). Eggshell quality improved with increasing dietary levels of calcium due to increased calcium retention and calcium balance. Increasing levels of dietary putrescine did not have a negative effect on eggshell quality; however, calcium intake was lower at higher supplemented levels of putrescine. It was observed that dietary calcium in excess of requirements resulted in increased egg production and eggshell quality. Eggshell quality improved when hens were fed 3.5% calcium diet in combination with 0.10% putrescine. It was concluded that small supplements of dietary putrescine may improve eggshell quality, depending on dietary calcium concentration. PMID- 11885905 TI - Effect of physical feed restriction during rearing on large white turkey breeder hens: 1. Growth performance. AB - Large White turkey breeder hens were fed ad libitum (CC), feed-restricted from 16 to 24 wk (CR), feed-restricted from 3 to 16 wk (RC), or feed-restricted from 3 to 24 wk (RR). Feed restriction was implemented so that RC and RR hens achieved a 45% reduction in BW compared to CC hens at 16 wk. From 16 to 24 wk, feed was allotted to RR and CR hens to maintain a slight increase in BW. At the completion of each restriction period, hens were gradually released back to ad libitum feeding. At 30 wk of age, hens were photostimulated for a 20-wk summer season egg production cycle. Mean BW for all treatments were different (P < or = 0.05) at 16 and 30 wk. At the end of lay, hens on treatment CR were not different in BW from treatment CC hens, and treatment RR hens were not different in BW from treatment RC hens. Hens on treatment RR had the greatest BW gain and feed consumption leading into the production cycle. All treatment hens lost BW from the time of first egg until 47 wk of age. Hens on treatment CC lost significantly (P < or = 0.05) more relative BW (%) than those in any other treatment. Coefficient of variation for flock uniformity was similar for all treatments at time of photostimulation. At the end of the study, cumulative feed consumption was significantly less for restricted treatments: 86.5, 83.1, 75.8, and 70.7 kg/hen for treatments CC, CR, RC, and RR, respectively. PMID- 11885906 TI - Liver proteolytic activity in tannic acid-fed birds. AB - The influence of tannic acid in the rate of growth (BWG), feed intake, protein efficiency ratio, and liver proteolytic activities (cathepsin A and D) were measured in growing male chickens. These birds were fed ad libitum over a 15-d experiment on 20% protein standard diets containing heated soybean (control, C) as the main source of protein. Tannic acid (TA; 25 g/kg diet) was added to all diets, except the control. It has been found that in comparison to control-fed birds, TA-fed birds showed a significant reduction (P < 0.01) in BWG, protein efficiency ratio, and relative weight of liver, together with a significant increase (P < 0.01) in the activities of cathepsin A and D in liver. Addition of TA to the control diet had no significant effect on feed intake. The possible nature of these results is discussed. PMID- 11885907 TI - Effect of enzymatic and chemical treatments on feather solubility and digestibility. AB - This study was designed to evaluate the effect of chemical and enzymatic approaches to digest poultry feathers from dead birds for the purpose of recycling their nutrients as animal feeds. The experimental treatments were as follows: 1) control, 2) 24-h enzyme, 3) 24-h NaOH, 4) 2-h NaOH, and 5) 2-h NaOH and 24-h enzyme. The feather N solubilities of the control, 24-h enzyme, 24-h NaOH, 2-h NaOH, and 2-h NaOH and 24-h enzyme treatments were 0.91, 2.55, 78.83, 30.03, and 50.34%, respectively. The pepsin digestibilities of unsolubilized feather residues from the control, 24-h enzyme, 2-h NaOH, and 2-h NaOH and 24-h enzyme treatments were 4.67, 13.19, 55.83, and 59.08%, respectively. The in vitro amino acid digestibilities of the 2-h NaOH and 24-h enzyme treatment were significantly higher than the 24-h enzyme or 2-h NaOH (P < 0.05), except for alanine, whereas the 2-h NaOH treatment had significantly higher amino acid digestibility than the 24-h enzyme treatment (P < 0.05), except for methionine and histidine. Costs per kilogram of solubilized feather for the 24-h enzyme, 24 h NaOH, 2-h NaOH, and 2-h NaOH and 24-h enzyme treatments were $9.64, 4.72, 12.39, and 22.97, respectively. The results indicated that prolonged incubation with NaOH improved feather solubility, whereas further enzyme treatment after NaOH treatment increased feather solubility, pepsin digestibility, and in vitro amino acid digestibility. PMID- 11885908 TI - Betaine does not improve performance of laying hens when the diet contains adequate choline. AB - An experiment was conducted with Hy-Line W36 hens to determine possible benefits from adding betaine to the diet of commercial laying hens. There was no benefit from the substitution of betaine for choline as measured by egg production, egg weight, egg content, or weight gain. PMID- 11885909 TI - Genetics and nursing: the interface in education, research, and practice. AB - Genetics is affecting all of health care, including nursing. The way in which nurses think about planning health care must be seen now through a "genetic eye" or lens, and nurses must learn to "think genetically." While efforts to integrate genetics into nursing began in earnest in the early 1980s, this effort did not accelerate until the mid-1990s. Before nursing can fully incorporate genetic knowledge into education and practice in a meaningful way, the ways in which genetics will influence health care must be understood. The basic knowledge, skills, and attitudes needed by health professionals are discussed as well as their integration into education and practice. Opportunities for nursing research in genetics are presented as are possible directions. Recommendations for facilitating the integration of genetics into nursing education, practice, and research are also presented. PMID- 11885910 TI - Ethical aspects of genetic testing. AB - This article explores ethical concerns and emerging dilemmas associated with the proliferation of information resulting from the extraordinary advances in molecular genetics. It provides an overview of the ethical and legal challenges associated with predictive testing for inherited disease currently being addressed in the literature. Finally, it offers a framework of ethical principles that can be used to guide nurses and other practitioners in the appropriate application of research findings to the clinical practice setting. The ethical guidelines of self-determination, benefit-burden ratio, and justice promulgated in The Belmont Report are interpreted in the new context of predictive genetic testing. The author concludes by discussing how to balance the technical imperative to advance genetic knowledge for the sake of human health with the ethical imperative to preserve the fundamental rights and liberties of both individuals and communities who are its recipients. PMID- 11885911 TI - Advancing genetic nursing research. PMID- 11885912 TI - Prevention of birth defects: folic acid. AB - Neural tube defects (NTDs) comprise an important category contributing to infant mortality. While some NTDs may be due to identifiable inherited or specific environmental factors, most are multifactorial, with genetic and environmental factors contributing to their occurrence. Folic acid has been found to have a protective effect against the recurrence and occurrence of NTDs. In addition to natural dietary sources, in the United States, all enriched grain products now are fortified with folic acid. In addition, all women who could become pregnant are recommended to consume 0.4 mg of folic acid daily. Despite these measures, not all women of childbearing age have added sufficient folic acid to their diets or take a vitamin supplement. Challenges remain regarding educating women of childbearing age about the potential health benefits of adequate folic acid consumption. PMID- 11885913 TI - Relationships of cortisol, perceived stress, genitourinary infections, and fetal fibronectin to gestational age at birth. AB - The authors investigated the role of stress and cortisol with patients having preterm labor (PTL) and preterm birth (PTB). The relationships of maternal cortisol, perceived stress, fetal fibronectin (fFN), and genitourinary infections to PTL and PTB were studied. A prospective, longitudinal, observational study (n = 78) was conducted in a private practice in central Texas. Subjects had 4 blood draws for cortisol measurements grouped by 15-19, 20-22, 23-26, 27-30, and 31-35 weeks of gestation. Subjects had 2 vaginal swabs forfFN, chlamydia, and bacterial vaginosis screens at 23-26 and 27-30 weeks with assessment of psychosocial stress at 23-26 and 31-35 weeks. Statistical analysis was by analysis of variance, Pearson correlations, Fisher exact test, and logistic regression. There were no significant differences between the PTB, PTL, and term groups on cortisol levels at any of the gestational periods. Cortisol concentrations at any gestational stage did not correlate with gestational age at birth. A relationship of cortisol to race was observed when comparing Caucasians to other ethnic groups. A correlation (r = 0.42, P < 0.001) between the change in Perceived Stress Scale (PSS) score and gestational age was observed. The greater the decrease in PSS scores, the longer was the gestational age. A significant increase in cortisol at 19-21 weeks (P < 0.04), 23-26 weeks (P < 0.05), and 31-35 weeks (P < 0.01) was observed in patients having genitourinary infection. PTL was also significantly increased in subjects having positive genitourinary infections at either 23-26 weeks or 27-30 weeks (P < 0.01). The sensitivity of fFN to predict PTL collected at 27-30 weeks was 40%, specificity 86%, positive predictive value 55%, and negative predictive value 83%. These results indicate that cortisol is a poor predictor of either PTL or PTB. A decrease in perceived stress during the 2nd trimester was associated with an increase in length of gestation, suggesting the possibility of stress reduction as an appropriate intervention for lengthening gestational age. PMID- 11885914 TI - From ecology to base pairs: nursing and genetic science. AB - With the mapping of the human genome has come the opportunity for nursing research to explore topics of concern to the maintenance, restoration, and attainment of genetic-related health. Initially, nursing research on genetic topics originated primarily from physical anthropology and from a clinical, disease-focused perspective. Nursing research subsequently focused on psychosocial aspects of genetic conditions for individuals and their family members. As findings emerge from current human genome discovery, new programs of genetic nursing research are originating from a biobehavioral interface, ranging from the investigations of the influence of specific molecular changes on gene function to social/ethical issues of human health and disease. These initiatives reflect nursing's response to discoveries of gene mutations related to phenotypic expression in both clinical and community-based populations. Genetic research programs are needed that integrate or adapt theoretical and methodological advances in epidemiology, family systems, anthropology, and ethics with those from nursing. Research programs must address not only populations with a specific disease but also community-based genetic health care issues. As genetic health care practice evolves, so will opportunities for research by nurses who can apply genetic concepts and interventions to improve the health of the public. This article presents an analysis of the evolution of genetic nursing research and challengesfor the future. PMID- 11885915 TI - Selected mechanisms of genotoxic effects of inorganic arsenic compounds. AB - Chronic exposure to inorganic arsenic compounds is responsible for the prevalance of various tumors, as well as of other diseases. A major problem is the exposure to inorganic arsenic (i-As) in drinking water that affects millions of people, primarily in Asia and South America. In these regions, the concentration of arsenic in drinking water amounts to several thousand microg/l and considerably exceeds the standard of 50 microg/l, recommended by the US Environmental Protection Agency. It is interesting that not all populations are equally sensitive to i-As. Therefore, the existing standard should be verified and the environmentally safe i-As concentration should be established. Bearing this in mind, it would be helpful to know the mechanisms of toxicity of inorganic arsenic compounds. In vitro and in vivo studies and examination of people exposed to high concentrations of i-As in drinking water show its genotoxicity. Inorganic As increases the frequency of micronuclei, chromosome aberrations and sister chromatid exchanges both in humans and in animals, but it does not induce point mutations. If arsenic does not affect DNA directly, then what is the mechanism of its toxicity? The results of various studies suggest that it may intensify toxic effects of other physical and chemical agents, especially by DNA repair inhibition. Besides, it is believed that inorganic arsenic compounds may cause changes in the cell redox potential and alter DNA methylation and phosphorylation of cell-cycle control proteins. Some data also suggest that i-As increases celluar proliferation and apoptosis. The purpose of this work is to present some views on cytotoxic mechanisms of inorganic arsenic compounds. PMID- 11885916 TI - Neurological and neurophysiological examinations of workers occupationally exposed to manganese. AB - The nervous system is the major target of the toxic effect of manganese (Mn) and its compounds. Nowadays, neurological diagnostics is directed towards early detection of symptoms and abortive forms, and the cases of serious damage of the nervous system are no longer reported. The aim of the present study was to assess the effects of manganese on the functions of the nervous system in workers exposed to this metal in the ship and electrical industries. The study covered a selected group of 75 male workers (mean age 39.17 yr +/- 9.79; range 20-56 yr), including 62 welders and fitters, as well as 13 workers involved in the battery production. Their employment duration ranged between 1 and 41 yr (mean 17.5 yr +/ 10.81). During the welding process the air Mn concentrations varied from 0.004 to 2.67 mg/m3 (arithmetic mean, 0.399 mg/m3; geometric mean, 0.154 mg/m3; standard deviation, 0.586). Of the 62 workers, 30 worked in the area with exceeding MAC value of 0.3 mg/m3. At the battery production workposts, Mn concentrations fell within 0.086-1.164 mg/m3 (arithmetic mean, 0.338 mg/m3, geometric mean, 0.261 mg/m3; standard deviation, 0.292). The values of current Mn exposure in the study group fell within the range below 0.01 and 2.67 mg/m3 (arithmetic mean, 0.4 mg/m; geometric mean, 0.15 mg/m3). Of the 13 subjects, 6 worked at the Mn air concentration exceeding MAC values. In the exposed group, the values of cumulated exposure index ranged from 0.008 to 35.52 (arithmetic mean, 8.045; geometric mean, 4.615; standard deviation, 6.562). The control group consisted of 62 men non-occupationally exposed to Mn, matched by sex, age and work shift distribution. Clinically, the increased emotional irritability, dysmnesia, concentration difficulties, sleepiness and limb paresthesia predominated among the disorders of the nervous system functions in workers chronically exposed to manganese. Neither in the central nor in the peripheral nervous system, the objective examinations revealed organic lesions that could provide grounds for diagnosing toxic encephalopathy or polyneuropathy. Generalized and paroxysmal changes were the most common recordings in the abnormal electroencephalography. Visual evoked potentials examinations showed abnormalities in the response evoked, which could be a signal of the optic neuron disorders and their significant relationship with cumulated exposure. The results of the study demonstrate that Mn exposure within the range of <0.01-2.67 mg/m3 (arithmetic mean, 0.4 mg/m3; geometric mean, 0.15 mg/m3) induces subclinical effects on the nervous system. PMID- 11885917 TI - Cancer risk among male farmers: a multi-site case-control study. AB - Farmers may experience exposure to several hazardous substances, and cancer risk in this occupational group is considered an important public health issue. In order to examine the association between cancer and farming among male agricultural workers, a hospital-based case-control study was conducted in five Italian rural areas. The cancer sites selected for the study were: lip, oral cavity and oropharynx, oesophagus, stomach, colon, rectum, lung, skin melanoma, skin non-melanoma, prostate, bladder, kidney, and non-Hodgkin's lymphoma. In all, 1525 newly diagnosed cases, aged 20-75 years, were ascertained in hospital records, covering the period between March 1990 and September 1992, and for 1279 of them, a detailed exposure information was collected by a standard questionnaire. Data analyses were performed comparing each cancer site to a control group, including a subset of the other cancer sites in the study. Unconditional logistic regression models were used in the statistical analyses. Increased risks of cancer associated with agricultural work were found for stomach (OR = 1.4, 95%CI:0.9-2.0), rectum (OR = 1.5, 95%CI:0.8-2.7), larynx (OR = 1.4, 95%CI:0.8-2.5), and prostate (OR = 1.4, 95%CI:1.0-2.1). The excess of prostate cancer was specifically related to application of pesticides (OR = 1.7, 95%CI:1.2-2.6). PMID- 11885918 TI - Assessment of the ability of health care providers to treat and prevent adverse health effects of pesticides in agricultural areas of Tanzania. AB - A survey of Tanzanian health care providers in agricultural areas was undertaken in 1991-1994 to assess their knowledge of toxic effects of pesticides, experiences and practices, as well as of their needs for appropriate information in order to develop effective strategies for reducing pesticide poisoning. Face to-face interviews were conducted with 104 physicians, clinical officers and nurses at health care facilities in the coffee and cotton growing areas. Eighty percent of respondents reported to have seen one and nine of them two to four cases of pesticide poisoning in the preceding three months. A significantly higher annual number of poisonings were observed in coffee than in cotton area (GM 0.5 vs 0.1). Also the number of cases registered in hospitals was considerably higher than that in the out-patient health care (GM 1.7 vs 0.2). Pesticide poisoning was regarded as a major problem in the community by 63% of health care providers, including 77% of hospital staff. One third of health care providers thought that a certain percent of pesticide poisoning cases remain unrecognized, and that this percentage is higher in cotton than in coffee growing areas. The respiratory tract was the major route for pesticide to enter the human body; this was followed by gastrointestinal tract, skin, and eyes. Only one percent of the respondents could identify the groups of pesticides (organophosphate vs organochlorine) mostly used in the study areas. The survey indicated that training of hospital staff in toxicity of pesticide exposure is an important task and a prerequisite for efficient recognition, diagnosis and treatment of pesticide poisoning cases in Tanzania. PMID- 11885919 TI - Prevalence, incidence and risk factors of carpal tunnel syndrome in a large footwear factory. AB - The study was conducted to assess the prevalence and incidence of carpal tunnel syndrome (CTS) in a large modern footwear factory and to identify factors predictive of CTS. To this end, 199 workers were examined in 1996, and 162 of them were re-examined in 1997. Ergonomic and psychosocial risk factors of CTS were assessed by workpost analysis and self-administered questionnaire. The prevalence of CTS at baseline in 1996 and in 1997 was 16.6% (95%CI: 11.4-21.7) and 11.7% (95%CI: 6.7-16.8), respectively. The incidence rate of CTS in 1997 was 11.7% (95%CI: 6.7-7.8). No specific type of job performance was associated with CTS. Obesity (OR = 4.4; 95%CI: 1.1-17.1) and psychological distress at baseline (OR = 4.3; 95%CI: 1.0-18.6) were strongly predictive of CTS. Rapid trigger movements of the fingers were also predictive of CTS (OR = 3.8; 95%CI: 1.0-17.2). A strict control of thework by superiors was negatively associatedwith CTS (OR = 0.5; 95%CI: 0.2-1.3). The prevalence and incidence of CTS in this workforce were largely higher than in the general population and numerous industries. The study highlights the role of psychological distress in workers exposed to a high level of physical exposure and psychological demand. PMID- 11885920 TI - The effect of sodium fluoride on the adenine nucleotide pool in erythrocytes of Wistar rats. AB - The effect of sodium fluoride on the content of adenine nucleotides, adenine nucleotide pool and energy potential of erythrocytes was studied in male Wistar rats, depending on the dose and time of exposure. Sodium fluoride was administered for 4 and 8 weeks at 4 or 16 ppm through a gastric tube. The concentration of fluorine in serum, ATP, ADP and AMP content in blood and erythrocytes, adenine nucleotide pool and energy potential of erythrocytes were calculated. The results were expressed in SI units and compared statistically with Student's t-test (Statgraphics v. 5.0 software). A significant reduction in the content of ATP and ADP and an increase in the content of AMP in erythrocytes was found after 4 weeks of exposure to 4 or 16 ppm NaF. The adenine nucleotide pool and energy potential were reduced with the smaller dose. After 8 weeks, the ADP content remained significantly reduced with the smaller dose, while the greater dose was associated with a higher energy potential of the cells. Correlations between serum concentration of fluorine, content of adenine nucleotides and adenine nucleotide pool in erythrocytes were noted in all study groups. PMID- 11885921 TI - The effect of heavy metals on the immune system at low concentrations. AB - The present study describes the effect of cadmium on lymphokines that cannot be directly traced to an allergen, or antigen in order to be able to explain various immunological processes. Exposure to various environmental pollutants is known to induce epithelial and inflammatory changes, characterized by a release of cytokines and other soluble mediators. Heavy metals like CdCl2 can induce, or inhibit the synthesis and expression of the inflammatory cytokines IL-1beta, IL 4, IL-6, TNF-alpha, IFN-gamma and ICAM-1. Normal human peripherial blood mononuclear cells (PBMCs) were exposed for different periods of times (1 to 96 h) to 0, 5, 25 and 50 micromoles CdCl2, and mRNA for the above cytokines was quantified by RT-PCR. Highly purified blood B cells and PBMCs from healthy donors were stimulated with IL-4 and aCD40 mAb and incubated with non-toxic concentrations of cadmium chloride (0,1-10 micromol). Levels of IgG and IgE were measured in the supernatants. Proliferation and expression of surface markers were determined by measuring [3H]-thymidine incorporation and by flow cytometry. The study showed that the in vitro synthesis of IgE by purified B cells or PBMCs stimulated with IL-4/aCD40 is inhibited by Cd at doses as low as 0,1 microM. Cd was found to inhibit IL-4/aCD40 induced proliferation of purified B cells and PBMCs in a dose dependent fashion. Most strikingly, only IgE but not IgG synthesis of purified B cells was inhibited by Cd. These data suggest that inhibition of IgE synthesis in human B lymphocytes by Cd seems to be a selective effect on immunoglobulin synthesis. PMID- 11885922 TI - Soil remediation--an alternative to abate human exposure to heavy metals. AB - This paper addresses the issue of soil pollution in the context of historical background, and its implications for the human exposure to heavy metals. The importance of metal bioavailability is also stressed. Various approaches to the problem are described, including administrative and technical preventive measures. PMID- 11885924 TI - Simulations in health risk assessment. AB - Health risk assessment procedure provides a clear and systematic form of quantitative (or semi-quantitative) description of environmental health impact. It is well known that this approach is burdened with various types of uncertainties of different origin and nature. Therefore, the results of risk assessment should always contain both the "number" and the "measure of uncertainty". The problem is that even if one does attempt to take account of the uncertainty, one does not know a priori what is the probability of getting a given risk value within the specified range of uncertainty. A promising tool for the assessment of risk which provides a means of describing the sensitivity with respect to different exposure factors and evaluating different intervention scenarios is the technique of Monte Carlo simulation. In this probabilistic approach all variables and parameters used in risk assessment may be regarded as distributions throughout the analysis. A process of repeated simulations is then used, during which the estimated quantity (risk in this case) is calculated many times (usually 10,000 or more) with randomly chosen values of variables and parameters, covering their range of variability and reproducing the assumed distribution density. The final result is given in the form of a probability distribution of risk. The idea of Monte Carlo simulations in health risk assessment concerning the exposure to heavy metals in drinking water is illustrated in the population living in the vicinity of the "Lubna" waste site, taken as an example. PMID- 11885923 TI - Nutrient intake patterns in gastric and colorectal cancers. AB - The purpose of the study was to present the dietary risk pattern in gastric and colorectal cancers, using the same methodological approach in a parallel hospital based case-control study. In all, 180 cases of colorectal cancer and 80 cases of stomach cancer, confirmed histopathologically, were enrolled from the University Hospital in Cracow. A high intake of carbohydrates was associated with an increased risk of colorectal cancer (OR = 2.45). For stomach cancer, a moderate consumption of carbohydrates markedly increased relative risk (OR = 4.29), while a high intake of carbohydrates increased the risk by 8.73. The patterns of dietary risk factors related to intake of fats were definitively different in both cancer sites. The higher fat consumption was not associated with the higher risk of stomach cancer. A medium intake of fats increased the risk of colorectal cancer by 1.96 and that above 83 g/day by 2.20. In colorectal cancer, the significant protective effect of retinol, carotene and vitamin C has been evidenced, however, only carotene and vitamin E were inversely correlated with stomach cancer. PMID- 11885925 TI - Magnetic fields classified as potentially carcinogenic. PMID- 11885926 TI - Avian and human influenza a virus strains possess different intracellular nucleoprotein oligomerization efficiency. AB - We have previously shown (Prokudina-Kantorovich EN and Semenova NP, Virology 223, 51-56, 1996) that the nucleoprotein (NP) of influenza A virus forms in infected cells oligomers which in the presence of SDS and 2-mercaptoethanol (ME) as reducing agent are stable at room temperature (RT) and dissociate at 100 degrees C. Here we report that the efficiency of intracellular NP oligomerization depends on the host origin of influenza A virus strain. Thus, in the cells infected with avian influenza A virus strains the viral NP was almost completely oligomerized and only traces of monomeric NP were detected by polyacrylamide gel electrophoresis (PAGE) in unboiled samples. However, in the cells infected with human influenza A virus strains, besides oligomeric NP also a significant amount of non-oligomerized monomeric NP was detected in unboiled samples. In purified virions of avian and human strains the same difference in NP monomers/oligomers ratio was detected as in the infected cells. A reassortant having all internal protein genes from a human strain and the glycoprotein genes from an avian strain revealed the same intracellular pattern of NP monomers/oligomers ratio as its parental human virus. These findings suggest that the type of NP oligomerization is controlled by the NP gene. The possible connection between the accumulation of protease-sensitive monomeric NP in cells infected with a human influenza strain and the parallel accumulation of cleaved NP in these cells is discussed. PMID- 11885927 TI - Activity of anthocyanins from fruit extract of Ribes nigrum L. against influenza A and B viruses. AB - Earlier, we have detected antiviral activity in an extract from Ribes nigrum L. fruits ("Kurokarin", name of the one species of black currant in Japanese) against influenza A and B viruses, and herpes simplex virus 1 (Knox et al., Food Processing 33, 21-23, 1998). In the present study, the antiviral activity of constituents of a Kurokarin extract and the mechanism of its antiviral action were examined. Kurokarin extracts were separated to fractions A to D by column chromatography. The major constituents of the fraction D were estimated as anthocyanins. The fraction D was further fractionated by thin-layer chromatography (TLC) to fractions A' to G'. The fraction E' consisted of 3-O alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl-cyanidin and 3-O-beta-D glucopyranosyl-cyanidin, and the fraction F' consisted of 3-O-alpha-L rhamnopyranosyl-beta-D-glucopyranosyl-delphinidin and 3-O-beta-D-glucopyranosyl delphinidin, identified by high performance liquid chromatography (HPLC) with standards and by high resolution mass spectrometry. The fractions D' to G' showed potent antiviral activity against influenza viruses A and B. The additive antiviral effect of a combination of the fractions E' and F' was assessed. Anthocyanins in the fraction F' did not directly inactivate influenza viruses A and B, but they inhibited virus adsorption to cells and also virus release from infected cells. PMID- 11885928 TI - Nucleotide sequence and phylogenetic analysis of a segment of a highly virulent strain of infectious bursal disease virus. AB - The complete nucleotide sequences encoding precursor polyprotein (VP2-VP3-VP4) and VP5 of a highly virulent (hv) infectious bursal disease virus (IBDV), UPM97/61 was determined. Comparison of the deduced amino acid sequences with the published ones revealed 8 common amino acid substitutions, which were found only in the hv IBDV including the UPM97/61 strain. Three of the amino acid substitutions (222 Ala, 256 Ile and 294 Ile) were used as a marker for determining hv IBDV strains. The other five substitutions (685 Asn, 715 Ser, 751 Asp, 990 Val and 1005 Ala) were also conserved in hv IBDV strains isolated in various countries. UPM97/61 strain demonstrated also 8 unique amino acid substitutions of which 3 were in VP2, 4 in VP3 and 1 in VP4. There was 1 unique amino acid substitution in VP5 at position 19 (Asp-->Gly) not found in other strains. However, all the strains have a conserved 49 Arg. The amino acid sequence of UPM97/61 strain differed by 1.09% from the Japanese (OKYM) and Hong Kong (HK46) strains, and by 1.48% from the Israeli (IBDVKS) and European (UK661) strains. Hence, UPM97/61 is more closely related to the hv strains from Asia. However, phylogenetic analysis indicated that the origin of UPM97/61 might be the same as that of other hv strains isolated from other parts of the world. PMID- 11885929 TI - Analysis of the extracellular processing of HIV-1 gp160-derived peptides using monoclonal antibodies specific to H-2Dd molecule complexed with p18-I10 peptide. AB - The immunodominant peptide of human immunodeficiency virus 1 gp 160 for murine cytotoxic T cells of H-2d haplotype, has been originally identified as a 15 amino acid residue peptide P18IIIB (RIQRGPGRAFVTIGK) (Takahashi et al., 1988). Further studies have indicated that a more active form of the peptide is generated by removal of the C-terminal dipeptide by angiotensin-I-converting enzyme (ACE), and additional detailed studies have shown that the actual immunodominant peptide is a decamer P18-I10 (RGPGRAFVTI) (Kozlowski et al., 1993). The effect of proteolytic processing on the antigenicity of P18IIIB peptide and its analogs was investigated by functional T cell assays based on the ability of T cell receptor (TCR) to recognize a specific major histocompatibility complex class I (MHC I)/peptide complex. Recently we described a new monoclonal antibody (MAb) KP15 directed against the MHC-I molecule H-2Dd complexed with the 10-mer peptide P18 I10. Using this MAb, the cell surface H-2Dd/P18-I10 complex can be easily detected by flow cytometry (Polakova et al., 2000). Here we examined whether peptides longer than P18-I10 decamer form H-2Dd complexes recognized by KP15 MAb. Further we also analyzed how the ACE processing of P18IIIB-related peptides of different length affects their ability to form complexes with H-2Dd recognized by MAb KP15. These experiments confirmed that the ACE digestion of 15-mer peptide P18IIIB is the most effective in the production of a peptide capable of forming complex with H-2Dd recognized by KP15 MAb. The ACE digestion of longer peptides (16-mer to 19-mer) did not produce a significant quantity of peptides, capable of forming H-2Dd complexes recognizable with by MAb KP15. Peptides shorter than P18IIIB (13-mer to 10-mer), notably the optimally sized P18-I10 peptide lost most of their capacity to form H-2Dd complexes recognized by KP15 MAb. Our results show that the extracellular processing of MHC-I-restricted peptides, which cannot be overlooked in designing peptide-based vaccines, can be also studied by as simple and rapid assay as flow cytometry, provided MAbs specific to a particular MHC-I/peptide complex are available. PMID- 11885930 TI - Development of a thermoresistant tissue culture rinderpest vaccine virus. AB - The currently used Plowright's tissue culture rinderpest vaccine (RBOK strain) gives full protection and lifelong immunity, but it is highly thermolabile and requires maintenance of cold chain from vaccine production till delivery. Keeping in view the need for a thermostabile vaccine in tropical developing countries with limited refrigeration facilities, we passaged serially the RBOK strain of rinderpestvirus (RPV) at gradually elevated temperature up to 40 degrees C to obtain a thermoresistant RPV (TR-RPV) mutant. The thermoresistance (thermostability) and antigenicity of TR-RPV were compared with those of the vaccine virus by various methods, confirming the acquired properties. Thus, the infectivity titres of the TR-RPV mutant and vaccine virus were determined after incubation for various times at 37 degrees C. Regression analysis indicated that TR-RPV had a half-life of 1.81 hr and a degradation constant of 0.1656, while the parent vaccine virus had a half-life of 1.11 hr and a degradation constant of 0.2686. In capture ELISA with four different monoclonal antibodies (MAbs) to the N protein of RPV, TR-RPV showed a 10-fold higher reactivity with one MAb as compared to the vaccine virus. Although TR-RPV did react also with the other three MAbs, its reactivity was only 4-5 times higher than that of the vaccine virus. A treatment of the virus with Triton X-100 resulted in 2-4 times higher reactivity with the MAbs. The 35S-methionine-labeled vaccine virus-and TR-RPV infected Vero cell lysates showed 6 polypeptide bands with identical pattern of migration in polyacrylamide gel electrophoresis in the presence of SDS (SDS PAGE). Radioimmunoprecipitation assay (RIPA) of the TR-RPV and vaccine virus with a rabbit anti-RPV immune serum (RHIS) and bovine anti-RPV hyperimmune serum (BHIS) showed the presence of four identical antigenic proteins, namely H, N, F and M, for both viruses. It can be concluded that TR-RPV has indeed retained the antigenic properties of the parental vaccine virus besides acquiring thermoresistance. PMID- 11885931 TI - Early gene expression of vaccinia virus strains replicating (Praha) and non replicating (modified vaccinia virus strain Ankara, MVA) in mammalian cells. AB - Modified vaccinia virus Ankara strain (MVA) is a safe highly attenuated non pathogenic virus suitable as a vector for developing various vaccines. Study of expression of a reporter beta-galactosidase gene under the control of an early vaccinia virus (VV) promoter in MVA and non-modified vaccinia virus Praha strain showed that early transcription in MVA is elevated in comparison with non modified VV. This property was demonstrated in various cell cultures including CV1 cells, human lung diploid cells, chicken primary fibroblasts but not in bone marrow-derived mouse dendritic cells. There the relationship between the elevated early transcription and the permisivity of cells for MVA was not observed. PMID- 11885932 TI - Nucleotide sequences of the coat protein and readthrough protein genes of the Chinese GAV isolate of barley yellow dwarf virus. AB - The nucleotide sequences of the coat protein (CP) and readthrough protein (RTP) genes of the Chinese GAV isolate of Barley yellow dwarf virus (BYDV) were determined. The CP and RTP genes of GAV isolate comprised 600 and 1374 nucleotides, respectively. When the CP and RTP gene sequences of GAV isolate were compared with those of BYDV isolates MAV-PS1, P-PAV, NY-SGV and Cereal yellow dwarf virus RPV (CYDV-RPV), the highest similarity (97.2%) between the CP genes of GAV and MAV-PS1 isolates was observed, while the RTP genes of these two isolates shared a lower similarity (87.8%). The results of the alignment of the deduced amino acid sequences of RTP showed that the sequence diversity observed was located at the C terminus. PMID- 11885933 TI - A modified immunofluorescence assay for detection of Japanese encephalitis virus infected cells. AB - Japanese encephalitis virus (JEV) infections are currently detected by indirect immunofluorescence (IF) assay using virus-specific antibodies on acetone-fixed smears. On few occasions, the acetone treatment was reported to damage certain epitopes on JE virus (JEV) glycoprotein. Here, we have made an attempt to adopt quick paraformaldehyde fixation followed by a short detergent treatment of cells in suspension for identification of JEV-infected brain cells of laboratory-reared Toxorhynchitis splendens mosquito larvae using virus-specific antibodies. JEV positive cells could be scored by the presence of a well defined intracellular immunofluorescence staining against unstained uninfected antibody-treated cells. The advantage of this assay is that stained cell suspensions can be stored for up to 4 weeks, allowing analysis at convenience. Thus, the modified IF assay can be employed as an additional/alternate technique to standard IF assay for detection of JEV in cells and also to screen hybridoma cell lines for anti-JEV antibody production. PMID- 11885934 TI - Significance of molecular identification of hepatitis C virus RNA in diagnosis of cryptogenic hepatitis in children. AB - Viral etiology of hepatitis is routinely proved by standard immunological tests detecting specific antibodies. However, identification of specific antibodies cannot always be conclusive. Since specific hepatitis C virus (HCV) antibodies may appear after some months of the infection, identification of HCV RNA and/or hepatitis G virus (HGV) RNA should clarify the etiology of hepatitis. The aim of this study was to diagnose etiologically unknown hepatitis by a reverse transcription-polymerase chain reaction (RT-PCR) testing of the presence of HCV RNA and HGV RNA. The study involved 33 children with histologically proved hepatitis. The presence of HCV and any signs of autoimmune disease were not observed at the beginning of the follow-up study. During 2.5 years of the follow up HCV-RNA was found in the blood and liver biopsies in 17 patients. Eight of them became HCV antibodies-positive during the follow-up. None of them eliminated the virus from the blood during the follow-up. In two other patients HCV-RNA was found only in the liver. HGV infection in all cryptogenic patients was excluded by PCR testing. Identification of HCV RNA RT-PCR allowed to diagnose 19 out of 33 (57.6%) patients with cryptogenic hepatitis. The etiology of the hepatitis in remaining 12 patients has to be established. PMID- 11885935 TI - Hepatitis G virus co-infection may affect the elimination of hepatitis C virus RNA from the peripheral blood of hemodialysis patients. AB - Hemodialysis patients are at risk for hepatitis C virus (HCV) and hepatitis G virus (HGV) infection. The aim of this study was to investigate the possible influence of HGV co-infection on HCV RNA elimination from the peripheral blood of hemodialysis patients. The study involved 144 persons, all with HCV antibodies and HCV RNA. Among 144 patients 24 (16.7%) were positive for HGV RNA. After 2.5 years of observation 80 patients (55.6%) were still HCV RNA-positive. In the latter group 18 patients were co-infected with HGV and 62 were HGV RNA-negative. During 2.5 years of the follow-up study 64 patients eliminated HCV RNA from the serum. In this group only 6 patients were HGV co-infected. None of the HGV positive patients eliminated HGV RNA from the serum. The higher incidence of HGV co-infection in the group of patients who remained HCV RNA-positive (18/80, 22.5%), in comparison to the group of HCV antibodies-positive patients who lost HCV in the blood (6/64, 9.4%, P < 0.0001) suggests, that the co-infection with HGV may delay the spontaneous elimination of HCV RNA from the blood. PMID- 11885936 TI - Control over HIV-1 replication by an antibiotic; a novel vaccination strategy with a drug-dependent virus. PMID- 11885938 TI - Kaposi's sarcoma associated herpesvirus (KSHV) or human herpesvirus 8 (HHV8). PMID- 11885937 TI - Epstein-Barr virus. PMID- 11885939 TI - Comments on the phylogenetics and evolution of herpesviruses and other large DNA viruses. PMID- 11885940 TI - Therapeutic vaccination. PMID- 11885941 TI - DNA mucosal HIV vaccine in humans. AB - This is a demonstration of immune activation by delivery of genetic vaccines in human mucosa. We analyzed the local and systemic responses in HIV-1 infected individuals following intraoral jet-injections of HIV-1 DNA constructs encoding the nef, rev, and tat regulatory genes. The immunological responses of the oral mucosa may be representative of other mucosal sites and was therefore selected for induction of mucosal reactivity by DNA immunization. The oral and intramuscular routes induced specific systemic T cell proliferative responses. Immunohistochemical analysis of oral biopsies 2 days after immunization revealed increased levels of granulocytes and T cells as well as expression of HLA-DR. T cell markers for CD3 +, CD4 + and CD8 + were significantly increased in the vaccinated mucosa. Vaccine-specific local and systemic antibodies were present during the immunization. However, increases were neither seen in the local or systemic titer nor in the B-cell accumulation in response to the immunizations. The presence of HIV-1 plasmid DNA was observed in mucosal biopsies as well as local proinflammatory T lymphocyte immune responses with predominantly IL-2 expression in the oral mucosal transudate. PMID- 11885942 TI - Anti-influenza therapies. PMID- 11885943 TI - Synthesis of influenza virus: new impetus from an old enzyme, RNA polymerase I. AB - Reverse genetics systems, i.e., systems for the generation of virus entirely from cloned cDNA, have been established for most nonsegmented negative-sense RNA viruses. In contrast, the generation of influenza A viruses (whose genome is composed of eight segments of negative-sense RNA) was not possible until 1999, likely due to the inherent technical difficulties of providing all eight viral RNAs as well as the four viral proteins required for replication and transcription. In 1999, we (Neumann et al., 1999, Proc. Natl. Acad. Sci. USA 96, 9345-9350) and others (Fodor et al., 1999, J. Virol. 73, 9679-9682) demonstrated the generation of influenza A virus from plasmids, relying on the cellular enzyme RNA polymerase I for the synthesis of influenza viral RNAs. In this review, we provide background on RNA polymerase I transcription and discuss its use for the generation of influenza virus from cloned cDNAs. PMID- 11885944 TI - Comparative pathogenesis of HBV and HCV. PMID- 11885945 TI - In vitro models for hepatitis C. PMID- 11885946 TI - Brief overview on cellular virus receptors. PMID- 11885947 TI - Emerging/disappearing viruses future issues concerning polio eradication. PMID- 11885948 TI - Emergence and selection of RNA virus variants: memory and extinction. AB - Two features of viral quasispecies are reviewed: the presence of memory genomes as minority components of their mutant spectra, and viral extinction due to enhanced mutagenesis. Memory has been documented with several genetic markers of the important animal picornavirus foot-and-mouth disease virus (FMDV). The presence of memory genomes in viral quasispecies may accelerate their adaptive response whenever a selective constraint has already been experienced by a viral population during previous stages of its evolution. Enhanced mutagenesis has been shown to lead to losses of infectivity of a number of RNA viruses: poliovirus, vesicular stomatitis virus, human immunodeficiency virus type 1 and FMDV. These observations, based on the theoretical prediction of the existence of a copying error-threshold for maintenance of genetic information, may contribute to the development of a new antiviral strategy. PMID- 11885949 TI - Quantification and genotyping in management of chronic hepatitis B and C. PMID- 11885950 TI - Prions--role of the peripheral nervous system. PMID- 11885951 TI - Bornaviruses. PMID- 11885952 TI - Rabies. PMID- 11885953 TI - DNA-chip technology and infectious diseases. PMID- 11885954 TI - Importance of hemagglutinin glycosylation for the biological functions of influenza virus. PMID- 11885955 TI - Folding of viral glycoproteins in the endoplasmic reticulum. AB - The endoplasmic reticulum (ER) is a subcellular compartment specialized in folding and assembly of newly synthesized polypeptides. The polypeptides expressed in the ER include all secretory proteins produced in the cell, lumenal or membrane-bound proteins of the endocytic/vacuolar and secretory compartments and transmembrane proteins that operate at the plasma membrane. In the lumen of the ER, molecular chaperones and folding factors facilitate the maturation of newly synthesized proteins. In a process defined as ER-quality control, they also warrant that only properly structured and assembled products leave the ER and are transported to their target organelles and compartments. If proper maturation fails, the aberrant products are degraded. Quality control in the ER is essential to prevent exit of improperly regulated or not-functional products that could lead to harmful effects. The mechanisms of protein folding and quality control in the ER are far from being fully understood. They are fundamental for the life of cells and organisms, but they are also linked to important human hereditary diseases in which mutated gene products are retained in the ER and degraded (e.g., cystic fibrosis and hereditary lung emphysema). PMID- 11885956 TI - The utility of H-2 class I knockout mice. PMID- 11885957 TI - High resolution structural studies of complex icosahedral viruses: a brief overview. AB - Structural descriptions of viral particles are key to our understanding of their assembly mechanisms and properties. We will describe the application of X-ray crystallography and electron cryomicroscopy to the structural determination of the bluetongue virus core and the herpesvirus capsid. These represent the highest resolution structural studies carried out by these techniques on such complex and large icosahedral virus particles. The bluetongue virus core consists of two layers of distinct proteins with different protein packing symmetries, while the herpes virus capsid is made up of four types of proteins with 3.3 MDa per asymmetric unit. The structural results reveal that each of these proteins has distinct folds and they are packed uniquely to form stable particles. PMID- 11885958 TI - HIV-1 Tat vaccines. PMID- 11885959 TI - Differential binding mode of diverse cyclooxygenase inhibitors. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are competitive inhibitors of cyclooxygenase (COX), the enzyme that mediates biosynthesis of prostaglandins and thromboxanes from arachidonic acid. There are at least two different isoforms of the enzyme known as COX-1 and -2. Site directed mutagenesis studies suggest that non-selective COX inhibitors of diverse chemical families exhibit differential binding modes to the two isozymes. These results cannot clearly be explained from the sole analysis of the crystal structures of COX available from X-ray diffraction studies. With the aim to elucidate the structural features governing the differential inhibitory binding behavior of these inhibitors, molecular modeling studies were undertaken to generate atomic models compatible with the experimental data available. Accordingly, docking of different COX inhibitors, including selective and non-selective ligands: rofecoxib, ketoprofen, suprofen, carprofen, zomepirac, indomethacin, diclofenac and meclofenamic acid were undertaken using the AMBER program. The results of the present study provide new insights into a better understanding of the differential binding mode of diverse families of COX inhibitors, and are expected to contribute to the design of new selective compounds. PMID- 11885960 TI - Prediction of enantiomeric selectivity in chromatography. Application of conformation-dependent and conformation-independent descriptors of molecular chirality. AB - In order to process molecular chirality by computational methods and to obtain predictions for properties that are influenced by chirality, a fixed-length conformation-dependent chirality code is introduced. The code consists of a set of molecular descriptors representing the chirality of a 3D molecular structure. It includes information about molecular geometry and atomic properties, and can distinguish between enantiomers, even if chirality does not result from chiral centers. The new molecular transform was applied to two datasets of chiral compounds, each of them containing pairs of enantiomers that had been separated by chiral chromatography. The elution order within each pair of isomers was predicted by means of Kohonen neural networks (NN) using the chirality codes as input. A previously described conformation-independent chirality code was also applied and the results were compared. In both applications clustering of the two classes of enantiomers (first eluted and last eluted enantiomers) could be successfully achieved by NN and accurate predictions could be obtained for independent test sets. The chirality code described here has a potential for a broad range of applications from stereoselective reactions to analytical chemistry and to the study of biological activity of chiral compounds. PMID- 11885961 TI - Computer-aided design of chiral ligands. Part I. Database search methods to identify chiral ligand types for asymmetric reactions. AB - The utility of database searching to identify chiral ligand motifs is outlined. The key elements of three known chiral ligands have been described as bond vectors. The CAVEAT program was then used to screen the Cambridge Structural Database (CSD), portions of the Chemical Abstracts Services three-dimensional database (CAS-3D), and the TRIAD tricyclic structure database for scaffolds containing these elements. Scaffolds corresponding to the known starting points were identified indicating that this method can be used to identify chiral ligand structural motifs. In addition, alternate structural motifs were found that suggested alternative possible ligands. PMID- 11885962 TI - MOSBY: a molecular structure viewer program with portability and extensibility. AB - A molecular structure viewer program, MOSBY has been developed for studies that use atomic coordinates to understand the structures of protein molecules. The program is designed to be portable with a comprehensive user interface by our high-throughput graphics library. In addition, it cooperates with extension modules customized for individual research topics and analysis. For example, an electron density module loads and displays electron density maps derived in X-ray crystallographic analysis superimposed to an atomic model. A molecular dynamics module reads a trajectory file of the results of molecular dynamics calculations and animates the structure. These plug-in modules are devised to function without modification to the MOSBY program. For variations of analysis and calculations with atomic coordinates, the portability and extensibility illustrated by MOSBY play an important rule in scientific computational tools with active software development. PMID- 11885963 TI - A novel approach for identifying the surface atoms of macromolecules. AB - A significant number of atoms lie buried beneath the "molecular surface" of proteins and other biologic macromolecules. Interactions between ligands and these macromolecules are dominated by interactions with the "surface atoms". Although interactions with the "buried" or interior atoms of the macromolecule certainly contribute to the total intermolecular interaction energy, many computer-assisted drug design (CADD) strategies can benefit from the identification of those atoms "on the surface" of proteins and other macromolecules. We have developed a simple, yet novel method to distinguish the surface atoms of macromolecules from the interior atoms which is based on computing the atomic contributions to the solvent-accessible surface (SAS) area. This report describes that method and demonstrates that it compares very favorably with four alternative methods. PMID- 11885964 TI - Physiological and pharmacological role of lysophosphatidic acid as modulator in mechanotransduction. AB - The mechanotransduction mechanism is believed to play an important role in maintenance of cellular homeostasis in a wide variety of cell types. In particular, the mechanotransduction system in vascular endothelial cells may be an essential mechanism for local hemodynamic control. Elevations in intracellular free Ca2+ concentration ([Ca2]i) are an important signal in the initial step of mechanotransduction and mechanosensitive (MS) cation channels are thought to be a putative pathway; however, the molecular mechanisms remain unclear. We found that lysophosphatidic acid (LPA), a bioactive phospholipid, sensitizes the response of [Ca2+]i to mechanical stress in several cell types. Employing real-time confocal microscopy, local increases in [Ca2+]i in several regions within the cell during application of mechanical stress were clearly visualized in bovine lens epithelial and endothelial cells in the presence of LPA. The phenomenon was termed "Ca2+ spots". Pharmacological studies revealed that Ca2+ spots arise due to influx through MS channels. In this report, our data indicating the possible significance of LPA as an endogenous factor involved in regulation of mechanotransduction is reviewed. Furthermore, our findings suggest that the Ca2+ spot is a novel phenomenon occurring as an elementary Ca2+-influx event through MS channels directly coupled with the initial step in mechanotransduction. PMID- 11885965 TI - Hepatoprotective drugs for the treatment of virus-induced chronic hepatitis: from hypercarcinogenic state to hypocarcinogenic state. AB - Interferon (IFN)-based therapy is a standard treatment for chronic hepatitis caused by hepatitis C virus (HCV) infection. This treatment is effective in approximately 30-40% of the patients and using ribavirin in combination with IFN increases the rate of sustained virologic clearance. For the remaining patients, glycyrrhizin is often used. Glycyrrhizin is known to prevent the development of hepatocellular carcinoma (HCC), but glycyrrhizin is usually administered intravenously. Drugs that are effective by oral administration are convenient for patients for long-term administration, and development of more effective drugs than glycyrrhizin is preferable. However, studies on drugs for the treatment of hepatitis are not actively conducted, and promotion of the study of drugs in this area is encouraging. For that reason, we show our approach to study drugs for the treatment of hepatitis. We analyzed the effect of glycyrrhizin on hepatitis as a standard chemical using the mouse liver injury model. Based on this, we screened drugs and found that a coumarin derivative seems to be one of model chemicals for the treatment of hepatitis. PMID- 11885966 TI - Adenylate cyclase/protein kinase A signaling pathway enhances angiogenesis through induction of vascular endothelial growth factor in vivo. AB - We previously reported that endogenous prostaglandins (PGs) may increase cAMP facilitated angiogenesis through the induction of vascular endothelial growth factor (VEGF) in rat sponge implantation models. In the present experiment, we tested whether or not adenylate cyclase / protein kinase A (AC/PKA)-dependent VEGF induction enhanced angiogenesis in this model. Topical daily injections of 8 bromo-cAMP enhanced angiogenesis in a dose-dependent manner. Forskolin, an activator of AC, also facilitated angiogenesis as did amrinone, an inhibitor of phosphodiesterase. VEGF induction was confirmed by the increased levels in the fluids in the sponge matrix after topical injection of 8-bromo-cAMP. Immunohistochemical investigation further revealed the VEGF-expressed cells in the sponge granulation tissues to be fibroblasts, and the intensity of positive reactions was enhanced by 8-bromo-cAMP, forskolin and amrinone. Angiogenesis without topical injections of the above compounds was suppressed by SQ22,536, an inhibitor for AC, or H-89, an inhibitor for PKA, with concomitant reductions in VEGF levels. Daily topical injections of neutralizing antibody or anti-sense oligonucleotide against VEGF significantly suppressed angiogenesis. PGE2-induced angiogenesis was suppressed with SQ22,536 or H-89. These results suggested that AC/PKA-dependent induction of VEGF certainly enhanced angiogenesis and that pharmacological tools for controlling this signaling pathway may be able to facilitate the management of conditions involving angiogenesis. PMID- 11885967 TI - Assessment of affinity and dissociation ability of a newly synthesized 5-HT2 antagonist, AT-1015: comparison with other 5-HT2 antagonists. AB - This study investigated the binding affinities of a newly synthesized 5-HT2 antagonist, AT-1015 (N-[2-[4-(5H-dibenzo[a,d]cyclohepten-5-ylidene) piperidino]ethyl]-1-formyl-4-piperidinecarboxamide monohydrochloride monohydrate) for [3H]ketanserin bindings to 5-HT2 receptors in the rabbit cerebral cortex membranes using the radioligand binding assay method. The affinity of this compound was also compared with other 5-HT2-selective antagonists such as ketanserin, sarpogrelate, cyproheptadine and ritanserin, and the results showed that AT-1015 has a high pKi value for the 5-HT2 receptor. The rank order of these antagonists are: ritanserin > ketanserin approximately equal to AT-1015 > cyproheptadine approximately equal to sarpogrelate. We also evaluated the dissociation ability (slow or rapid) of AT-1015 in the rabbit cerebral cortex membrane and compared it with other 5-HT2 antagonists using the radioligand binding assay method. The blockade of [3H]ketanserin binding sites in the rabbit cerebral cortex induced by ketanserin and sarpogrelate was readily reversed by washing, whereas the inhibition by AT-1015, cyproheptadine and ritanserin was not readily reversed by washing. The % of control after washing are 76.10% and 49.55% for AT-1015 at 10(-7.5) and 10(-7.0) M, 67.32% and 50.17% for cyproheptadine at 10(7.5) and 10(-7.0) M, and 72.38% and 39.80% for ritanserin at 10(-9.5) and 10( 9.0) M concentrations, respectively. Thus, these findings suggest that AT-1015 has antagonistic properties towards the 5-HT2 receptor and also shows that AT 1015 slowly dissociates from the 5-HT2 receptor, whereas, ketanserin and sarpogrelate dissociate rapidly from the 5-HT2 receptor, which do not correlate with their affinity. PMID- 11885968 TI - Deterioration of spatial learning performances in lipopolysaccharide-treated mice. AB - It is well demonstrated that acute or chronic stress leads to reduction of learning ability. Lipopolysaccharide (LPS), a component of the outer membrane of gram-negative bacteria, induces profound physiological and behavioral changes, including fever, decrease in food motivation, and decrease in social behavior. These changes might be interpreted as an acute stress reaction to the LPS. In the present study, therefore, we investigated the effects of LPS (400-800 microg/kg, i.p.) on spatial learning performances using C57BL/6J male mice. In the Morris water-maze task, spatial learning performances were examined in six trials of training for two consecutive days. LPS-treated mice took a longer time to reach the hidden platform than control mice (F(1,60)=4.80801, P<0.05 at 600 microg/kg). In addition, injection of LPS decreased the percent of correct choices in the Y maze test (P<0.05 at 800 microg/kg). LPS, however, did not alter the body weight, grip tone, motor activity or swimming speed. Taken together, these results indicate that LPS treatment specifically impaired spatial learning performances. PMID- 11885969 TI - Activation of a potassium conductance by extracellular alkaline pH in oocytes of Xenopus laevis. AB - Electrophysiological properties of Xenopus oocytes exposed to alkaline extracellular pH (pHo) were investigated by measuring whole-cell currents using the two-electrode voltage-clamp method. Alkaline pHo (8.5-10.5) elicited an outward current in a pHo-dependent manner with a concomitant increase in the membrane conductance. This outward-current response was dependent on K+ because it was suppressed by a K+ channel blocker tetraethylammonium+ (20 mM), and the reversal potential of the response was in good agreement with the Nernst equation for K+. The response was not affected by pretreatment of oocytes with the acetoxymethyl ester of bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (10,uM), a membrane-permeant intracellular Ca2+ chelator, but it was augmented by forskolin (0.4 microM), a stimulant of adenylate cyclase. The outward-current response originates in the oocyte but not in the surrounding follicle cells because the current could still be evoked when follicle cells were removed by collagenase or when gap junctions connecting the oocyte membrane and follicle cells were blocked by 1-octanol (1 mM). It is concluded that the outward current elicited by alkaline pHo in Xenopus oocytes is dependent on the activation of K+ channels via the cAMP pathway and that the outward current originates in the oocyte rather than the surrounding follicle cells. PMID- 11885970 TI - Effects of calcium antagonists on the nitrergic nerve function in canine corpus cavernosum. AB - Effects of calcium antagonists on nitrergic nerve function were examined in the isolated canine corpus cavernosum. In the cavernous strips precontracted with phenylephrine, transmural electrical stimulation elicited frequency-dependent (2 5 Hz) relaxations that were abolished by N(G)-nitro-L-arginine (10(-5) M), a nitric oxide (NO) synthase inhibitor; 1H[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ, 10(-6) M), a soluble guanylate cyclase inhibitor; and tetrodotoxin (3 x 10( 7) M). The relaxations were not affected by treatment with nifedipine or nicardipine (10(-8) - 10(-6) M), L-type specific calcium channel inhibitors, but were significantly inhibited by amlodipine or cilnidipine, inhibitors of L- plus N-type calcium channels, in a concentration-related manner (10(-7) - 10(-6) M). All of the inhibitors used did not affect the relaxations induced by exogenous NO (acidifed NaNO2). These findings suggest that N-type, but not L-type, calcium channels are responsible for increasing cytosolic free calcium, a prerequisite for the synthesis of NO, in the nitrergic dilator nerves innervating the corpus cavernosum. PMID- 11885971 TI - A novel analgesic compound OT-7100 attenuates nociceptive responses in animal models of inflammatory and neuropathic hyperalgesia: a possible involvement of adenosinergic anti-nociception. AB - We studied the effects of OT-7100 (5-n-butyl-7-(3,4,5 trimethoxybenzoylamino)pyrazolo [1,5-a]pyrimidine), a novel analgesic compound, on the inhibitory action of adenosine on the contraction of guinea pig ileum and investigated the effects of OT-7100 on the nociceptive responses in animal models of inflammatory and peripheral neuropathic hyperalgesia and decreases spinal c Fos expression. OT-7100 at 0.3 - 3 microM significantly enhanced the inhibitory effect of adenosine on the contraction of guinea pig ileum. The efficacy of OT 7100 (1, 3 or 10 mg/kg, p.o.) on hyperalgesia induced by yeast or substance P and in the Bennett model was significantly suppressed by coadministration of the adenosine A1 antagonist DPCPX (0.01 or 0.1 pmol/animal, i.t.), while OT-7100 without DPCPX significantly increased the nociceptive threshold in each rat model. OT-7100 (3, 10 and 30 mg/kg per day, p.o.) significantly inhibited the mechanical nociceptive threshold in the injured paw in the Chung model. OT-7100 (30 mg/kg, p.o.) significantly decreased the number of Fos-LI neurons in the spinal dorsal horn in the Bennett model. These finding suggest that OT-7100 inhibits hyperalgesia in these animal models possibly by enhancing adenosinergic neurotransmission in the dorsal horn, although we still lack direct evidence for it. PMID- 11885973 TI - A comparative study of the fluoroquinolone antibacterial agents on the action potential duration in guinea pig ventricular myocardia. AB - We examined the effects of ten fluoroquinolone antibacterial agents, levofloxacin, sitafloxacin, trovafloxacin, ciprofloxacin, gemifloxacin, tosufloxacin, gatifloxacin, grepafloxacin, moxifloxacin and sparfloxacin, on action potentials recorded from guinea pig ventricular myocardia. Sparfloxacin prolonged action potential duration (APD) by about 8% at 10 microM and 41% at 100 microM. Gatifloxacin, grepafloxacin and moxifloxacin also prolonged APD at 100 microM by about 13%, 24% and 25%, respectively. In contrast, levofloxacin, sitafloxacin, trovafloxacin, ciprofloxacin, gemifloxacin and tosufloxacin had little or no APD-prolonging effect at concentrations as high as 100 microM. These findings suggest that there are differences in potency to prolong QT interval among the fluoroquinolones. PMID- 11885972 TI - The influence of the protein kinase A system in differentiation of HL-60-Eo cells to eosinophils induced by histamine. AB - The influence of the protein kinase A (A kinase) system in differentiation of HL 60-Eo cells to eosinophils induced by histamine was studied. Although 8-Cl-cAMP caused inhibitions of proliferation and [3H]thymidine uptake of HL-60-Eo cells similarly to histamine, no significant eosinophilic differentiation was observed. Histamine as well as 8-Cl-cAMP caused elevation of A kinase activity. However, KT 5720, an inhibitor of A kinase, had no effect on histamine-induced eosinophil differentiation. RIalpha antisense oligodeoxynucleotide caused significant inhibition of HL-60-Eo cell growth, but RIIbeta antisense oligodeoxynucleotide had no effect. On the other hand, neither of the antisense oligodeoxynucleotides showed potentiating effects on growth inhibition induced by histamine. In addition, RIalpha and RIIbeta antisense oligodeoxynucleotides caused neither differentiation to eosinophils itself nor potentiation of histamine-induced differentiation. From these findings, it was concluded that A kinase is not correlated directly with differentiation of HL-60-Eo cells to eosinophils. PMID- 11885974 TI - Activation of background membrane conductance by the tyrosine kinase inhibitor tyrphostin A23 and its inactive analog tyrphostin A1 in guinea pig ventricular myocytes. AB - ABSTRACT-The effects of the tyrosine kinase (TK) inhibitor tyrphostin A23 and its inactive analog tyrphostin Al on background membrane conductance were investigated in guinea pig ventricular myocytes. TK-inhibiting A23 reversibly increased membrane conductance under conditions designed to minimize Na+, Ca2+, K+, and Na+-K+ pump currents. Similar stimulatory action was obtained with TK inactive Al. The tyrphostin-induced current was inhibited by omitting external Na+ or Ca+, suppressed by chelating internal Ca2+, blocked by external Cd2+ and Ni2+, and insensitive to changes in internal Cl- concentration. We conclude that tyrphostins have a direct, TK-independent action that increases membrane conductance probably by stimulating Na+-Ca2+ exchange. PMID- 11885975 TI - Ameliorative effects of azaindolizinone derivative ZSET845 on scopolamine-induced deficits in passive avoidance and radial-arm maze learning in the rat. AB - Effects of ZSET845 (3,3-dibenzylimidazo[1,2-a]pyridin-2-(3H)-one), a newly synthesized cognitive enhancer, and donepezil and tacrine on the scopolamine induced cognitive deficits in rats were examined in passive avoidance and radial arm maze tasks. ZSET845 (0.01 mg/kg) showed a greater ameliorative effect than donepezil (0.1 mg/kg) or tacrine (1 mg/kg) in the passive avoidance task. In the radial-arm maze task, ZSET845 (0.1 mg/kg) also showed a greater effect than donepezil (10 mg/kg) or tacrine (10 mg/kg). ZSET845 induced an increase in the choline acetyltransferase (ChAT) activity in the hippocampus, suggesting that the ameliorative effects of ZSET845 are related to the increase in the ChAT activity in the hippocampus. PMID- 11885976 TI - Residual dipolar couplings: synergy between NMR and structural genomics. AB - Structural genomics is on a quest for the structure and function of a significant fraction of gene products. Current efforts are focusing on structure determination of single-domain proteins, which can readily be targeted by X-ray crystallography, NMR spectroscopy and computational homology modeling. However, comprehensive association of gene products with functions also requires systematic determination of more complex protein structures and other biomolecules participating in cellular processes such as nucleic acids, and characterization of biomolecular interactions and dynamics relevant to function. Such NMR investigations are becoming more feasible, not only due to recent advances in NMR methodology, but also because structural genomics is providing valuable structural information and new experimental and computational tools. The measurement of residual dipolar couplings in partially oriented systems and other new NMR methods will play an important role in this synergistic relationship between NMR and structural genomics. Both an expansion in the domain of NMR application, and important contributions to future structural genomics efforts can be anticipated. PMID- 11885978 TI - A spin-state-selective experiment for measuring heteronuclear one-bond and homonuclear two-bond couplings from an HSQC-type spectrum. AB - Recently, a set of selective 1D experiments with spin-state-selective excitation for CH spin systems was introduced by Parella and Belloc (J. Magn. Reson., 148, 78-87 (2001)). We have expanded and generalized this concept further, and demonstrated that a very simple experiment utilizing spin-state-selective filtering can be used for simultaneous measurement of heteronuclear 1JNH (or 1JCH) and geminal 2JHH couplings from two-dimensional 15N-1H (or 13C-1H) correlation spectrum. The experiment has very high sensitivity owing to the preservation of equivalent coherence transfer pathways analogous to the sensitivity and gradient enhanced HSQC experiment. However, overall length of the pulse sequence is 1/(2J) shorter than the gradient selected SE-HSQC experiment. Furthermore, the spin-state-selection can be utilized between NH and NH2 (or CH and CH2) moieties by changing the phase of only one pulse. The pulse scheme will be useful for the measurement of scalar and residual dipolar couplings in wide variety of samples, due to its high sensitivity and artifact suppression efficiency. The method is tested on NH2 and CH2 moieties in 15N- and 15N/13C labeled ubiquitin samples. PMID- 11885977 TI - 1H-filtered correlation experiments for assignment and determination of coupling constants in backbone labelled proteins. AB - The implementation of [13Calpha,13C',15N,2Halpha] labelled amino acids into proteins allows the acquisition of high resolution triple resonance experiments. We present for the first time resonance assignments facilitated by this new labelling strategy. The absence of 1JCalpha,Cbeta couplings enables us to measure 1JCalpha,C' scalar and 1DCalpha,C' residual dipolar coupling constants using modified HNCA experiments which do not suffer from sensitivity losses characteristic for 13C constant time experiments. PMID- 11885980 TI - Structure refinement of flexible proteins using dipolar couplings: application to the protein p8MTCP1. AB - The present study deals with the relevance of using mobility-averaged dipolar couplings for the structure refinement of flexible proteins. The 68-residue protein p8MTCP1 has been chosen as model for this study. Its solution state consists mainly of three alpha-helices. The two N-terminal helices are strapped in a well-determined alpha-hairpin, whereas, due to an intrinsic mobility, the position of the third helix is less well defined in the NMR structure. To further characterize the degrees of freedom of this helix, we have measured the dipolar coupling constants in the backbone of p8MTCP1 in a bicellar medium. We show here that including D(dip)HN dipolar couplings in the structure calculation protocol improves the structure of the alpha-hairpin but not the positioning of the third helix. This is due to the motional averaging of the dipolar couplings measured in the last helix. Performing two calculations with different force constants for the dipolar restraints highlights the inconstancy of these mobility-averaged dipolar couplings. Alternatively, prior to any structure calculations, comparing the values of the dipolar couplings measured in helix III to values back calculated from an ideal helix demonstrates that they are atypical for a helix. This can be partly attributed to mobility effects since the inclusion of the 15N relaxation derived order parameter allows for a better fit. PMID- 11885979 TI - Solution structure of N-terminal SH3 domain of Vav and the recognition site for Grb2 C-terminal SH3 domain. AB - The three-dimensional structure of the N-terminal SH3 domain (residues 583-660) of murine Vav, which contains a tetra-proline sequence (Pro 607-Pro 610), was determined by NMR. The solution structure of the SH3 domain shows a typical SH3 fold, but it exists in two conformations due to cis-trans isomerization at the Gly614-Pro615 bond. The NMR structure of the P615G mutant, where Pro615 is replaced by glycine, reveals that the tetra-proline region is inserted into the RT-loop and binds to its own SH3 structure. The C-terminal SH3 domain of Grb2 specifically binds to the trans form of the N-terminal SH3 domain of Vav. The surface of Vav N-terminal SH3 which binds to Grb2 C-terminal SH3 was elucidated by chemical shift mapping experiments using NMR. The surface does not involve the tetra-proline region but involves the region comprising the n-src loop, the N terminal and the C-terminal regions. This surface is located opposite to the tetra-proline containing region, consistent with that of our previous mutagenesis studies. PMID- 11885981 TI - Position of residues in transmembrane peptides with respect to the lipid bilayer: a combined lipid Noes and water chemical exchange approach in phospholipid bicelles. AB - The model transmembrane peptide P16 (Ac-KKGLLLALLLLALLLALLLKKA-NH2) was incorporated into small unaligned phospholipid bicelles, which provide a 'native like' lipid bilayer compatible with high-resolution solution NMR techniques. Using amide-water chemical exchange and amide-lipid cross-relaxation measurements, the interactions between P16 and bicelles were investigated. Distinctive intermolecular NOE patterns observed in band-selective 2D-NOESY spectra of bicellar solutions with several lipid deuteration schemes indicated that P16 is preferentially interacting with the 'bilayered' region of the bicelle rather than with the rim. Furthermore, when amide-lipid NOEs were combined with amide-water chemical exchange cross-peaks of selectively 15N-labeled P16 peptides, valuable information was obtained about the position of selected residues relative to the membrane-water interface. Specifically, three main classes were identified. Class I residues lie outside the bilayer and show amide water exchange cross-peaks but no amide-lipid NOEs. Class II residues reside in the bilayer-water interface and show both amide-water exchange cross-peaks and amide-lipid NOEs. Class III residues are embedded within the hydrophobic core of the membrane and show no amide-water exchange cross-peaks but strong amide-lipid NOEs. PMID- 11885982 TI - A novel PH-cT-COSY methodology for measuring JPH coupling constants in unlabeled nucleic acids. application to HIV-2 TAR RNA. AB - A quantitative analysis of JPH scalar couplings in nucleic acids is difficult due to small couplings to phosphorus, the extreme overlap of the sugar protons and the fast relaxation of the spins involved in the magnetization transfer. Here we present a new methodology that relies on heteronuclear Constant Time Correlation Spectroscopy (CT-COSY). The three vicinal 3JPH3', 3JPHS' and 3JPHS" scalar couplings can be obtained by monitoring the intensity decay of the P1-H3'(i-1) peak as a function of the constant time T in a 2D correlation map. The advantage of the new method resides in the possibility of measuring the two 3JPH5' and 3JPH5" scalar couplings even in the presence of overlapped H5'/H5" resonances, since the quantitative information is extracted from the intensity decay of the P H3' peak. Moreover, the relaxation of the H3' proton is considerably slower than that of the H5'/H5" geminal protons and the commonly populated conformations of the phosphate backbone are associated with large 3JPH3' couplings and relatively small 3JPH5'/H5". These two facts lead to optimal signal-to-noise ratio for the P H3' correlation compared to the P-H5'/H5" correlation. The heteronuclear CT-COSY experiment is suitable for oligonucleotides in the 10-15 kDa molecular mass range and has been applied to the 30mer HIV-2 TAR RNA. The methodology presented here can be used to measure P-H dipolar couplings (DPH) as well. We will present qualitative results for the measurement of P-Hbase and P-H2' dipolar couplings in the HIV-2 TAR RNA and will discuss the reasons that so far precluded the quantification of the DPHS for the 30mer RNA. PMID- 11885983 TI - Characterization of polyacrylamide-stabilized Pfl phage liquid crystals for protein NMR spectroscopy. AB - A new polymer-stabilized nematic liquid crystal has been characterized for the measurement of biomolecular residual dipolar couplings. Filamentous Pf1 phage were embedded in a polyacrylamide matrix that fixes the orientation of the particles. The alignment was characterized by the quadrupolar splitting of the 2H NMR water signal and by the measurement of 1H-15N residual dipolar couplings (RDC) in the archeal translation elongation factor 1beta. Protein dissolved in the polymer-stabilized medium orients quantitatively as in media without polyacrylamide. We show that the quadrupolar splitting and RDCs are zero in media in which the Pf1 phage particles are aligned at the magic angle. This allows measurement of J and dipolar couplings in a single sample. PMID- 11885984 TI - The NMR structure of the class I human ubiquitin-conjugating enzyme 2b. PMID- 11885985 TI - MQ-hCN-based pulse sequences for the measurement of 13C1'-1H1', 13C1'-15N, 1H1' 15N, 13C1'-13C2', 1H1'-13C2',13C6/8-1H6/8, 13C6/8-15N, 1H6/8-15N, 13C6-13C5, 1H6 13C5 dipolar couplings in 13C, 15N-labeled DNA (and RNA). AB - A suite of multiple quantum (MQ) HCN-based pulse sequences has been developed for the purpose of collecting dipolar coupling data in labeled nucleic acids. All the pulse sequences are based on the robust MQ-HCN experiment which has been utilized for assignment purposes in labeled nucleic acids for a number of years and provides much-needed resolution for the dipolar coupling measurements. We have attempted to collect multiple couplings centered on the 13C1' and 13C6/8 positions. Six pulse sequences are described, one each for measurement of one bond 13C1'-1H1' and 13C6/8-1H6/8 couplings, one for measurement of one-bond 13C1' 15N and two-bond 1H1'-15N couplings, one for measurement of one-bond 13C6/8-15N and two-bond 1H6/8-15N couplings, one for measurement of one-bond 13C1'- 13C2' and two-bond 1H1'-13C2' couplings, and one for measurement of one-bond 13C6-13C5 and two-bond 1H6-13C5 couplings in the bases of C and T. These sequences are demonstrated for a labeled 18 bp DNA duplex in a 47 kDa ternary complex of DNA, CBFbeta, and the CBFalpha Runt domain, thus clearly demonstrating the robustness of the pulse sequences even for a very large complex. PMID- 11885987 TI - Complete 1H, 15N and 13C assignments of the carboxyl terminal domain of the ciliary neurotrophic factor receptor (CNTFR). PMID- 11885986 TI - Backbone sequential resonance assignments of yeast iso-2 cytochrome c, reduced and oxidized forms. PMID- 11885988 TI - 1H, 13C and '5N resonance assignments of GABARAP, GABAA receptor associated protein. PMID- 11885989 TI - Assignment of the 1H, 13C and 15N resonances of the catalytic domain of the rat 2',3'-cyclic nucleotide 3'-phosphodiesterase. PMID- 11885990 TI - A fistful of Euro. PMID- 11885991 TI - Research charities try pharma industry trick. PMID- 11885992 TI - Does rehabilitation have a place in oncology management? PMID- 11885993 TI - Germ-cell tumor survivors: the price for cure. PMID- 11885994 TI - Oestrogen/insulin-like growth factor-I receptor interaction in early breast cancer: clinical implications. AB - The expression of oestrogen receptor (ER) and that of the insulin-like growth factor-I receptor (IGF-IR) are positively correlated in breast cancer specimens. Their function is strongly linked in enhancing proliferative activity in normal and malignant human mammary epithelial cells in culture. This review examines the likely role of such a mechanism in the increased breast cancer risk reported in postmenopausal women with abdominal obesity. Precancerous breast lesions show an increased proportion of ER-positive cells with high proliferative activity, and recent studies suggest that genetic mutations or epigenetic variants in the ER alpha gene may increase the ER's sensitivity to oestrogen stimulation. Abdominal obesity in women is associated with higher concentrations both of free oestradiol and free IGF-I. Activation of their respective receptors may induce synergistic stimulation of mammary carcinogenesis. However, there is clinical evidence that progression in precancerous breast lesions may be delayed or reversed. Involution occurs spontaneously in a proportion of duct carcinoma in situ (DCIS) (intraductal) lesions as women approach the menopause, and antioestrogen therapy has been shown to reduce recurrence and progression of DCIS lesions. CONCLUSIONS: Intervention trials in breast cancer prevention would greatly benefit from surrogate response markers which could predict long-term benefit. Changes in ER and IGF-IR expression apart from those in standardised cytomorphological criteria, might predict the likelihood of DCIS involution in cancer prevention trials. Future studies could involve examination of serial core biopsies from normal breast tissue during trials of antioestrogens, retinoids or weight reduction interventions. Correlation of changes in these markers with changes in circulating IGF-I and oestradiol concentrations may help to clarify the roles of the markers. PMID- 11885995 TI - Second and subsequent lines of chemotherapy for metastatic breast cancer: what did we learn in the last two decades? AB - Despite almost 30 years of clinical cancer research, the true impact of second and subsequent lines of chemotherapy on the outcome of metastatic breast cancer patients, especially on the duration of survival, is still unknown. In the virtually incurable metastatic setting, issues like quality of life and patients' preferences gain particular relevance. At the turn of the century, in-depth rethinking of the design of clinical trials run in this challenging disease setting appears to be warranted. PMID- 11885996 TI - Quality of life and rehabilitation in social and professional life after autologous stem cell transplantation. AB - BACKGROUND: In this study, we report on quality of life (QOL) in long-term survivors after high-dose chemotherapy and autologous stem cell transplantation (ASCT) with special emphasis on rehabilitation in social and professional life. PATIENTS AND METHODS: The European Organization of Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ)-C30 questionnaire was sent by mail to 391 patients 1 to 12 years (median 31 months) after ASCT. The procedure was performed at our institution alone. Of the questionnaires 78% were returned and evaluated. RESULTS: Of the 238 patients who had an occupation and were employed, 132 (55%) have returned full time (68%) or part time (32%) to their previous occupation. A total of 139 patients (46%) received a 3- to 4-week inpatient rehabilitation treatment in specialised institutions following ASCT. Employment status post-transplantation and QOL were similar in these patients as compared with those who did not participate in rehabilitation programmes. Of the 304 evaluable patients, 39% reported physical problems that reduced their satisfaction with sex and intimacy. The general QOL was significantly reduced in the first year, improved with interval to transplant, and reached the level of the general population after 4 years. CONCLUSIONS: Our retrospective data showed that ASCT has a significant, unfavourable impact on QOL, including reintegration into social and professional life. Most symptoms and scores returned to normal after 3 to 6 years. Employment status and QOL were similar in patients who participated in a rehabilitation programme and those who did not. PMID- 11885997 TI - Canalicular stenosis secondary to weekly docetaxel: a potentially preventable side effect. AB - BACKGROUND: The purpose of this study was to describe canalicular stenosis as a mechanism for epiphora (excessive tearing) secondary to weekly docetaxel. PATIENTS AND METHODS: Fourteen patients with metastatic breast cancer who developed epiphora during weekly docetaxel therapy underwent an ophthalmologic examination, and probing and irrigation of the nasolacrimal ducts. The total duration of docetaxel therapy, the duration of treatment at the time of onset of epiphora, the number of infusions, the cumulative dose of docetaxel and the severity of canalicular stenosis were recorded. RESULTS: All 14 patients had anatomic narrowing of the canaliculi as the underlying mechanism for epiphora. Bicanalicular silicone intubation or dacryocystorhinostomy (DCR) was recommended for all 14 patients. Eleven patients underwent surgery and experienced resolution of their symptoms. The three patients who declined surgery continue to have epiphora at the time of this report. CONCLUSIONS: Canalicular stenosis is an underlying mechanism for epiphora in patients receiving weekly docetaxel. Bicanalicular silicone intubation should be considered early in the course of weekly docetaxel therapy to prevent complete closure of the canaliculi. Once complete or near complete stenosis of the canaliculi occurs, DCR with a permanent pyrex glass tube placement may become necessary to overcome the blockage of tear outflow. PMID- 11885998 TI - Long-term renal function after treatment for malignant germ-cell tumours. AB - OBJECTIVE: To evaluate prospectively renal function in patients with malignant germ-cell tumours (MGCTs) >10 years after retroperitoneal lymph node dissection alone (RPLND), radiotherapy alone (RAD) or different schedules of cisplatin-based chemotherapy with or without surgery/radiotherapy (CHEM). PATIENTS AND METHODS: In 85 patients, three groups were identified: RPLND, 14; RAD, 18; CHEM, 53, with subdivision of the latter group according to the cumulative cisplatin dose or the additional use of radiotherapy. Renal function was determined by 131Iodine Hippuran clearance or 99m DTPA glomerular filtration rate, and was assessed before treatment and four times during 14 years of follow-up. A value of <70% of the upper limit of the normal range identified impaired renal function. RESULTS: Twenty-five patients displayed long-term impaired renal function, 23 of them from the RAD or CHEM group. In the RAD group, renal function decreased by 8%, whereas a 14% reduction of renal function was observed in the CHEM group. In the CHEM group the cumulative dose of cisplatin, and in the RAD group the age at treatment, were associated with impairment of renal function. Combining all patients, age at treatment and the type of treatment were associated with impaired renal function. CONCLUSIONS: In 20-30% of the patients with germ-cell tumour, standard radiotherapy and chemotherapy strategies are followed by long term subclinical impaired renal function. These findings support current intentions to avoid overtreatment with these treatment modalities. PMID- 11886000 TI - Routine computerised tomographic scans of the thorax in surveillance of stage I testicular non-seminomatous germ-cell cancer--a necessary risk? AB - BACKGROUND: The standard management approach to stage I testicular non seminomatous germ-cell tumours (NSGCT) in the UK is a surveillance programme with adjuvant bleomycin, etoposide, cisplatin (BEP) chemotherapy being offered to individuals with high risk disease. Conventionally, computed tomography (CT) scanning of the thorax has formed part of the surveillance programme. This paper evaluates the contribution of routine thoracic CT imaging in the management of this disease. PATIENTS AND METHODS: We retrospectively reviewed the case notes of 168 patients with stage I NSGCT referred to the Wessex Medical Oncology Unit over a period of 13 years (1986-1998). These patients entered onto a surveillance programme that included serial chest X-ray follow up rather than thoracic CT. RESULTS: Forty-two out of 168 patients (25%) evaluated suffered relapse during the follow up period. Eight of 42 patients (19%) relapsed with intrathoracic disease. Seven out of eight of these patients (87.5%) had at least one other indicator of disease recurrence (elevated serum marker, abnormal abdominal CT). One of 42 patients (2.4%) relapsed with isolated intrathoracic disease with no other indicator of relapse. All patients with intrathoracic relapse had evidence of disease on chest X-ray. Of the 42 relapsing patients, 93% could be categorised as having good prognosis metastatic disease. Seven per cent relapsed with intermediate or poor prognostic disease; relapse in these patients would not have been detected earlier with the inclusion of routine thoracic CT. Only one patient has died giving a cure rate of 98% for relapsing patients. CONCLUSIONS: The elimination of chest CT did not compromise outcome but significantly reduced radiation exposure thereby minimising the risk of radiation-induced secondary malignancy. Continued review of surveillance programmes is essential if we are to optimise management of this disease. PMID- 11885999 TI - Evaluation of long-term toxicity in patients after cisplatin-based chemotherapy for non-seminomatous testicular cancer. AB - BACKGROUND: Because of the increasing number of long-term survivors of metastatic testicular germ-cell cancer, a general concern has been secondary morbidities, especially cardiovascular risk factors. PATIENTS AND METHODS: Thirty-two patients treated with cisplatin- and doxorubicin-containing chemotherapy > or = 13 years before the time of analyses were evaluated for neuro-, oto-, pulmonary-, vascular and gonadal toxicity including evaluation of myocardial damage and cardiovascular risk factors and analysis of microcirculation. RESULTS: Thirty percent of the patients showed abnormal left ventricle function. Elevated follicle stimulating hormone (FSH) and luteinising hormone (LH) levels in 75% of patients were often associated with low testosterone levels. Elevated total cholesterol levels were found in 82% and higher triglyceride levels in 44% of patients, most of them were overweight. About 25% of the patients developed diastolic arterial hypertension after chemotherapy. Reduced hearing was confirmed in 23% of patients, especially at frequencies higher than 3000 Hz. Moreover, 53% of patients presented transient evoked otoacoustic emissions. In 38% of patients non-symptomatic neuropathy was detected, in 28% symptomatic neuropathy, and in 6% disabling polyneuropathy. In 80% of patients with neuropathic symptoms additional morphological and functional abnormalities were found by nailfold capillary videomicroscopy, compared to only 57% of the patients without neuropathic symptoms. CONCLUSIONS: Patients cured by cisplatin-based chemotherapy for metastatic testicular cancer have to be cognizant of their unfavorable cardiovascular risk profile, that might be a greater risk than developing a relapse or second malignancy. PMID- 11886001 TI - Weekly chemotherapy with docetaxel, gemcitabine and cisplatin in advanced transitional cell urothelial cancer: a phase II trial. AB - PURPOSE: To evaluate the efficacy and toxicity of a combination of weekly docetaxel, gemcitabine and cisplatin in advanced transitional cell carcinoma (TCC) of the bladder. PATIENTS AND METHODS: Thirty-five chemotherapy-naive (adjuvant and neoadjuvant chemotherapy was allowed) patients with advanced TCC received intravenous docetaxel 35 mg/m2, gemcitabine 800 mg/m2 and cisplatin 35 mg/m2, on days 1 and 8 every 3 weeks. Prophylactic granulocyte-colony stimulating factor was given from days 3 to 6 and days 10 to 15, anti-emetics were used routinely. RESULTS: Most (27) patients (77.1%) had a performance status of 0 to 1 and eight (22.9%) had received prior adjuvant or neoadjuvant cisplatin-based chemotherapy. In the intention-to-treat analysis, the objective response rate was 65.6% [23/35 patients, 95% confidence interval (CI) 47.8% to 80.9%]. Ten patients (28.5%) achieved a complete response (95% CI 14.6% to 46.3%) and 13 (37.1%) a partial response (95% CI 21.5% to 55.0%). Median survival time was 15.5 months, median duration of response was 10.2 months and median time to progression was 8.9 months. Ten patients (28.5%) developed grade 3/4 neutropenia, including five (14.3%) who experienced febrile neutropenia, which was successfully treated. Grade 3/4 anaemia and thrombocytopenia occurred in 20% and 25.7% of patients, respectively; four patients required platelet transfusions. There were no treatment-related deaths. CONCLUSIONS: Weekly docetaxel, gemcitabine plus cisplatin is a highly effective treatment for chemotherapy-naive advanced TCC, and causes only moderate toxicity. This regimen should be considered as a suitable option that deserves further prospective evaluation through randomised phase III trials. PMID- 11886002 TI - Chemotherapy versus hormonal treatment in platinum- and paclitaxel-refractory ovarian cancer: a randomised trial of the German Arbeitsgemeinschaft Gynaekologische Onkologie (AGO) Study Group Ovarian Cancer. AB - BACKGROUND: The majority of patients with ovarian cancer are not cured by first line treatment. Until now, no study could demonstrate any substantial benefit when exposing ovarian cancer patients to second-line chemotherapy. However, most treatment regimens induce toxicity, thus negatively influencing the quality of rather limited life spans. Here we evaluate whether a second-line chemotherapy can offer any benefit compared with a less toxic hormonal treatment. PATIENTS AND METHODS: Patients with ovarian cancer progressing during platinum-paclitaxel containing first-line therapy or experiencing relapse within 6 months were eligible. Patients were stratified for response to primary treatment (progression versus no change/response), and measurable versus non-measurable disease. Treatment consisted of either treosulfan 7 g/m5 infused over 30 min or leuprorelin 3.75 mg injected subcutaneously or intramuscularly. Both regimens were repeated every 4 weeks. RESULTS: This study began in late 1996, and after 2.5 years accrual an interim analysis was performed when several investigators reported their concern about a suspected lack of efficacy. Following this analysis the recruitment was stopped early and the 78 patients already enrolled were followed up. The majority of patients received treatment until progressive disease was diagnosed or death occurred. Treatment delay was observed rarely and dose reduction was performed only in the treosulfan arm in 5% of 150 courses. Overall, both treatment arms were well tolerated. No objective responses were observed. The median survival time was 36 and 30 weeks in the treosulfan and leuprorelin arms, respectively. Overall survival did not differ between patients with relapse 3-6 months after first-line chemotherapy compared with patients with progressive disease within 3 months. CONCLUSIONS: The selected patient population represents a subgroup with extremely poor prognosis. Accordingly, results were not impressive. Both treatment arms showed favourable toxicity data, but failed to show remarkable activity, thus adding only limited evidence to the issue of whether patients with refractory ovarian cancer might benefit from second-line chemotherapy. Even stratified analysis did not identify any subgroup of patients in whom the administration of second-line chemotherapy could demonstrate a clinically relevant survival benefit. PMID- 11886003 TI - Multicentre phase II study of oxaliplatin as a single-agent in cisplatin/carboplatin +/- taxane-pretreated ovarian cancer patients. AB - BACKGROUND: This multicentre phase II open-label study evaluated safety and antitumour activity of oxaliplatin in cisplatin or carboplatin (cis/carboplatin) +/- taxane-pretreated advanced ovarian cancer (AOC) patients. PATIENTS AND METHODS: Forty-eight patients received oxaliplatin 130 mg/M2 intravenously every 3 weeks, 94% having a performance status (PS) 0-1. All were pretreated with cis/carboplatin and 21 (44%) with paclitaxel. The median number of involved organs was two, 18 (38%) had liver metastasis, 23 (48%) were platinum-resistant and 14 (29%) were taxane-resistant. Forty-two patients were evaluable for a response, 18 (43%) were platinum-resistant and 11 (26%) were taxane-resistant. RESULTS: A total of 253 cycles was administered (median: 5.5/patient). Median cumulative oxaliplatin dose was 666 mg/m2. National Cancer Institute-Common Toxicity Criteria toxicity analysis showed that seven patients (15%) had grade 3/4 thrombocytopenia, two patients (4%) had grade 3 neutropenia, and one patient had grade 3 anaemia. Eleven patients (23%) experienced grade 3 neurosensory toxicity. Of the 29 patients with peripheral neuropathy at the end of treatment, 55% had recovered or improved 1 month later. Eleven objective responses (two complete) were obtained in the 42 evaluable patients [ORR 26%, 95% confidence interval (CI) 14% to 42%], with 10/24 (42%, 95% CI 22% to 63%) in platinum sensitive, and 1 of 18 (5.6%, 95% CI 0% to 27%) in platinum-resistant patients. Median response duration was 9.2 months (95% CI 6.6% to 11.8%), and median progression-free and overall survival in all treated patients were 4.3 months (95% CI 3.0% to 5.7%) and 15.0 months (95% CI 11.1% to 18.8%), respectively. CONCLUSION: Oxaliplatin has a good safety profile and is active in cis/carboplatin +/- paclitaxel-pretreated AOC patients. PMID- 11886004 TI - Heated intra-operative intraperitoneal oxaliplatin after complete resection of peritoneal carcinomatosis: pharmacokinetics and tissue distribution. AB - PURPOSE: This article reports the pharmacokinetics (PK) of heated intra-operative intraperitoneal oxaliplatin and its tolerance profile. Oxaliplatin has demonstrated significant activity in advanced colorectal cancer, and this is the first publication concerning its intraperitoneal administration. METHODS: Twenty consecutive patients with peritoneal carcinomatosis (PC) of either gastrointestinal or uniquely peritoneal origin underwent complete cytoreductive surgery followed by intra-operative intraperitoneal chemo-hyperthermia (IPCH) with increasing doses of oxaliplatin. We performed IPCH using an open procedure (skin pulled upwards), at an intraperitoneal temperature of 42-44 degrees C, with 2 l/m2 of 5% dextrose instillate in a closed circuit. The flow-rate was 2 l/min for 30 min. Patients received intravenous leucovorin (20 mg/m2) and 5 fluorouracil (400 mg/m2) just before the IPCH to maximize the effect of oxaliplatin. We treated at least three patients at each of the six intraperitoneal oxaliplatin dose levels (from 260 to 460 mg/m2) before progressing to the next. We analysed intraperitoneal, plasma and tissue samples with atomic absorption spectrophotometry. RESULTS: The mean duration of the entire procedure was 8.4 +/- 2.7 h. Half the oxaliplatin dose was absorbed in 30 min at all dose levels. Area under the curve (AUC) and maximal plasma concentration (Cmax) increased with dose. At the highest dose level (460 mg/m2), peritoneal oxaliplatin concentration was 25-fold that in plasma. AUCs following intraperitoneal administration were consistently inferior to historical control AUCs after intravenous oxaliplatin (130 mg/m2). Intratumoral oxaliplatin penetration was high, similar to absorption at the peritoneal surface and 17.8 fold higher than that in non-bathed tissues. Increasing instillate volume to 2.5 l/m2 instead of 2 l/m2 dramatically decreased oxaliplatin concentration and absorption. There were no deaths, nor severe haematological, renal or neurological toxicity, but we observed two fistulas and three deep abscesses. CONCLUSIONS: Heated intraperitoneal chemotherapy gives high peritoneal and tumour oxaliplatin concentrations with limited systemic absorption. We recommend an oxaliplatin dose of 460 mg/m2 in 2 l/m2 of 5% dextrose for intraperitoneal chemo hyperthermia, at a temperature of 42-44 degrees C over 30 min. We may be able to improve these results by increasing the intraperitoneal perfusion duration or by modifying the instillate composition. PMID- 11886005 TI - Very young women (<35 years) with operable breast cancer: features of disease at presentation. AB - BACKGROUND: Breast cancer rarely occurs in young women. Our knowledge about disease presentation, prognosis and treatment effects are largely dependent upon older series. MATERIALS AND METHODS: We evaluated biological features and stage at presentation for 1427 consecutive premenopausal patients aged < or = 50 years with first diagnosis of invasive breast cancer referred to surgery at the European Institute of Oncology from April 1997 to August 2000. A total of 185 patients (13%) were aged < 35 years ('very young') and 1242 (87%) were aged 35-50 years ('less young'). The expression of estrogen receptors (ER), progesterone receptors (PgR), presence of vascular invasion (VI), grading (G), expression of Ki-67, HER2/neu overexpression, pathological stage according to TNM staging system (pTNM), pathological tumor size and number of axillary lymph node involvement were evaluated. RESULTS: Compared with less young patients, the very young patient group had a higher percentage of tumors classified as ER negative (P < 0.001), PgR negative (P = 0.001), higher expression of Ki-67 > or = 20% of cells stained; 62.2% versus 53%, (P < 0.001), vascular or lymphatic invasion (48.6% versus 37.3%, P = 0.006), and pathological grade 3 (P < 0.0001). There was no difference between the two groups for pT, pathological tumor size (pN) and number of positive lymph nodes. CONCLUSIONS: We conclude that compared with less young premenopausal patients, very young women have a greater chance of having an endocrine-unresponsive tumor, and are more likely to present with a higher grade, more extensively proliferating and vessel invading disease. Pathological tumor size, nodal status and number of positive axillary lymph-nodes have a similar distribution among the younger and the older cohorts, thus not supporting previous data indicating more advanced disease in younger patients at diagnosis of operable disease. PMID- 11886006 TI - Influence of alternate sequences of epirubicin and docetaxel on the pharmacokinetic behaviour of both drugs in advanced breast cancer. AB - BACKGROUND: Previously we observed a pharmacokinetic interference of epirubicin elimination when paclitaxel is given in combination in a sequence-dependent manner (i.e. when paclitaxel is administered as first drug). The aim of this study was to determine whether these sequence-dependent pharmacological effects were also evident when epirubicin was combined with docetaxel. PATIENTS AND METHODS: Patients who received epirubicin 75 mg/m2 or 90 mg/m2 as an intravenous bolus followed immediately by docetaxel 70 mg/m2 or 80 mg/m2 over a 1-h infusion, or the opposite sequence, every 3 weeks were eligible for this study. The pharmacokinetics of docetaxel, epirubicin and its metabolites were studied at the first and second cycle of treatment. Pharmacokinetic data were normalised to the lower dose of each drug. Toxicity was recorded at nadir and graded according to National Cancer Institute Common Toxicity Criteria. RESULTS: Twelve consecutive patients, each acting as their own control, entered the study. The sequence of drug administration of docetaxel and epirubicin did not affect the pharmacokinetics of the parent anthracycline. Statistically significant lower glucuronidation metabolism of epirubicin was observed in patients who received docetaxel before epirubicin. The pharmacokinetics of docetaxel were not influenced by the sequence of drug administration. No difference in haematological and non-haematological toxicity was observed in the two sequences of treatment. CONCLUSIONS: The pharmacokinetics of the parent anthracycline and of docetaxel were similar between the two schemes of treatment. The metabolic variations observed, i.e. differences in the plasma levels of epirubicin glucuronides, seem not to have clinical relevance. PMID- 11886007 TI - Phase II study of weekly docetaxel in patients with metastatic breast cancer. AB - BACKGROUND: This study was conducted to investigate the efficacy and toxicity of weekly docetaxel administration in patients with metastatic breast cancer. PATIENTS AND METHODS: Thirty-seven women were treated with 1 h infusions of docetaxel at 40 mg/m2/week after pre-medication with 8 mg dexamethazone. Each cycle consisted of three consecutive weekly treatments followed by a 1 week rest. All patients were assessed for toxicity; five patients were not assessable for clinical response, time to progression (TTP) and overall survival (OS) because of early treatment failure, but they were included in intention-to-treat analysis. RESULTS: Patients received a median of four cycles (range, 1-9), with a median dose intensity of 28 mg/m2/week (range 22-30) and a median relative dose intensity of 0.95 (range 0.73-1.0). No patients showed complete response, whereas 14 had partial response, which accounted for 38% of objective response rate [95% confidence interval (CI) 22% to 53%]. In addition, three patients (8%, 95% CI 0% to 17%) had stable disease over 6 months. Clinical responses were achieved at a median of three cycles (range 1-4 cycles). The median TTP and OS were 5 and 12 months, respectively. The weekly docetaxel regimen was generally well tolerated. About half of the patients experienced grade > or = 1 neutropenia; only 19% had grade 3/4 neutropenia, including one case of grade 4. No febrile neutropenia was observed and fluid retention syndrome was uncommon. Non-hematologic toxicity, however, such as asthenia/fatigue, nail damage, tearing or hearing disorders, was seen with successive treatment cycles. CONCLUSIONS: Weekly docetaxel at 40 mg/m2/week is an active and feasible regimen for patients with metastatic breast cancer. PMID- 11886008 TI - Neoadjuvant tamoxifen for hormone-sensitive non-metastatic breast carcinomas in early postmenopausal women. AB - BACKGROUND: From 1984 to 1996, 1581 postmenopausal women aged 50-70 years old were treated at Institut Bergonie for an infiltrative non-metastatic breast carcinoma with a positive estrogen and/or progesterone receptor determination. PATIENTS AND METHODS: Among them, 199 were treated with first line tamoxifen. Ninety-seven had operable disease (T2 >30 mm, T3, N0/1) and 102 had T4 tumours. RESULTS: After a mean treatment duration of 5.3 months, 89 T2 and T3 (92%) and 93 T4 (91%) were treated by surgery (conservative or not) with or without irradiation, or by irradiation alone. Conserving treatment levels were 53.6% and 44%, respectively. The other women were treated with either second-line chemotherapy or another hormonotherapy; the remaining patients continued regularly with tamoxifen. Overall survival is analysed with a 83 month median follow-up. CONCLUSIONS: A comparison between neoadjuvant endocrine therapy and surgery seems feasible to assess the concept of neoadjuvant hormonotherapy. PMID- 11886009 TI - Adjuvant chemotherapy in gastric cancer: 5-year results of a randomised study by the Italian Trials in Medical Oncology (ITMO) Group. AB - BACKGROUND: The aim of this study was to determine the efficacy of the EAP regimen (etoposide, adriamycin and cisplatin) followed by the Machover schedule (fluorouracil and folinic acid) given as adjuvant treatment to patients with poor prognostic factors (N+ or T3/4). PATIENTS AND METHODS: Before randomisation, the subjects were stratified on the basis of node involvement (N+ or N-) and the time from surgery to randomisation (< or = 21 days or > 22 days). The surgical procedures for sub-total or total gastrectomy with D2 dissection were standardised among the participating centres. RESULTS: Between December 1992 and December 1997, 274 patients were enrolled: 137 in the treatment arm and 137 in the control arm. The majority of the patients (90%) were N+. After a median follow up of 66 months (range 2-83), the 5-year overall survival (OS) was 52% in the treatment arm and 48% in the control arm [hazard ratio (HR) 0.93; 95% confidence interval (CI) 0.65-1.34]; the 5-year disease-free survival (DFS) was 49% and 44%, respectively (HR: 0.83; 95% CI 0.59-1.17). Among the patients with N /N+ (1-6), the 5-year OS was 61% in the treatment group and 60% in the control group; in those with N+ (1-6), it was 42% and 22%. The treatment was completed by 87% of patients. Drug-related grade 3/4 WHO toxicities included leukopenia (21%), nausea and vomiting (14%), mucositis (9%), neutropenia (3%) and thrombocytopenia (2%). There were two deaths due to sepsis. CONCLUSIONS: Although our results are not statistically significant, there was a limited relative risk reduction in the patients receiving adjuvant therapy (17% in DFS and 7% in OS). The data suggest that D2 surgery may have a favourable impact on OS. PMID- 11886011 TI - Is there an increased rate of additional malignancies in patients with mantle cell lymphoma? AB - PURPOSE: To examine the frequency of additional neoplasms preceding and following the diagnosis of mantle cell lymphoma (MCL). PATIENTS AND METHODS: A total of 156 patients with MCL treated on the hyperfractionated cyclophosphamide, vincristine, doxorubicin and dexamethasone alternated with methotrexate and cytosine arabinoside (Hyper-CVAD/M-A) program with or without rituximab from 1994 to 2000 were the subjects of this report. RESULTS: These patients were followed for a median time of 26 months, and a total of 32 (21%) additional neoplasms were diagnosed, 21 preceding the diagnosis of MCL and 11 following MCL. After excluding certain types of non-invasive neoplasms, including basal cell carcinoma, meningioma and cervical intraepithelial neoplasia, we observed seven second malignancies after the diagnosis of MCL, and the 5-year cumulative incidence rate of second malignancy was 11%. The observed-to-expected (O/E) ratio was 7/0.07 = 100 [95% confidence interval (CI) 49.3 to 186.6; P <0.0001]. Of the 21 malignancies diagnosed prior to MCL, 16 were invasive and five non-invasive. There were a total of 10 urologic malignancies occurring before or after the diagnosis of MCL was established. CONCLUSIONS: Our findings suggest that there is an increased incidence of second malignancies in patients with MCL. In addition, the high number of cases with urinary tract cancer in our series may substantiate prior reports describing a possible association between lymphoma and urologic malignancies. PMID- 11886010 TI - Clinical determinants of survival in patients with 5-fluorouracil-based treatment for metastatic colorectal cancer: results of a multivariate analysis of 3825 patients. AB - BACKGROUND: Patients with metastatic colorectal cancer are usually offered systemic chemotherapy as palliative treatment. A multivariate analysis was performed in order to identify predictors and their constellation that allow a valid prediction of the outcome in patients treated with 5-fluorouracil (5-FU) based therapy. PATIENTS AND METHODS: A total of 3825 patients treated with 5-FU within 19 prospective randomised and three phase II trials were separated into learning (n = 2549) and validation (n = 1276) samples. Data were analysed by tree analysis using the recursive partition and amalgamation method (RECPAM). A predictor could only enter the RECPAM analysis if the number of patients with missing values was < 33.3% within a node, and the minimal node size was set to 50 patients. Twenty-three potential predictors were grouped into subsets of laboratory variables (11 parameters), tumour-related variables (seven parameters) and clinical variables (five parameters). In the first step, tree analysis was performed separately for each predictor subset. The selected prognostic parameters of the resulting partial models (the 'winners') were entered into the general model. The classification rule from the data of the learning set was applied to the independent validation set. RESULTS: Winners of the subgroup analysis for laboratory variables were: platelets > or = 400 x 10(9)/l, alkaline phosphatase > or = 300 U/l, white blood cell (WBC) count > or = 10 x 10(9)/l and haemoglobin < 11 x 10(9)/l, and all predicted a worse outcome. Negative predictors within the subgroup of tumour parameters were: number of tumour sites more than one or more than two, presence of liver metastases or peritoneal carcinomatosis, which predicted a worse outcome. Furthermore, presence of lung metastases, a primary rectal cancer and presence of lymph node metastases all predicted a better outcome in the multivariate setting. Among the clinical parameters only performance status of ECOG 0 or 1 predicted better outcome. In the final regression tree, three risk groups could be identified: low risk group (n = 1111) with a median survival of 15 months for patients with ECOG 0/1 and only one tumour site; intermediate risk group (n = 904) with a median survival of 10.7 months for patients with ECOG 0/1 and more than one tumour site and alkaline phosphatase < 300 U/l or patients with ECOG > 1, WBC count < 10 x 10(9)/l and only one tumour site; high risk group (n = 534) with a median survival of 6.1 months for patients with ECOG 0/1 and more than one tumour site and alkaline phosphatase of > or = 300 U/l or patients with ECOG > 1 and more than one tumour site or WBC count > 10 x 10(9)/l. The median survival times for the good, intermediate and high risk groups in the validation sample were 14.7, 10.5 and 6.4 months, respectively. CONCLUSIONS: Patients can be divided into at least three risk groups depending on the four baseline clinical parameters: performance status, WBC count, alkaline phosphatase and number of metastatic sites. Any molecular or biological marker should be validated against these clinical parameters and decisions for more or less intensive treatments may be studied separately in these three risk groups. Also, clinical trials should be stratified according to the three risk groups. PMID- 11886012 TI - Rhabdomyosarcoma arising in mediastinal teratoma in an adult man: a case report. AB - We report a case of rhabdomyosarcoma which occurred in a mediastinal teratoma in a 44-year-old man. Presentation symptoms were chest pain, hoarseness and a cough. Diagnosis was fortuitous, performed by the histological and immunohistochemical study of a mediastinal tumour biopsy specimen that showed embryonal carcinoma and yolk sac tumour components associated with the rhabdomyosarcoma. After cisplatin based chemotherapy (bleomycin-etoposide-cisplatin), surgical resection of the residual mediastinal tumour was performed. Histological and immunohistochemical study of this tumour confirmed the presence of mature teratoma and embryonal rhabdomyosarcoma. Evolution was marked by a local extension of the mediastinal tumour, occurrence of multiple metastases and bone marrow involvement. The patient died 8 months after diagnosis despite chemotherapy and radiotherapy. A review of the literature reveals that the development of rhabdomyosarcoma in primary mediastinal teratomas is unusual in adults. The diagnostic, therapeutic and prognostic implications of such an association are reviewed. PMID- 11886013 TI - Significant impairment of high-dose methotrexate clearance following vancomycin administration in the absence of overt renal impairment. AB - BACKGROUND: Methotrexate is an antimetabolite cytotoxic drug which is predominantly renally excreted. Vancomycin, a glycopeptide antibiotic that is used in the febrile neutropaenic patient, can be nephrotoxic. There are no previous reports of any interactions between these two drugs. PATIENTS AND METHODS: We describe two patients with osteosarcoma treated with high-dose methotrexate-containing chemotherapy who had significantly delayed methotrexate clearance several weeks following exposure to vancomycin. RESULTS: These patients were treated with alternating chemotherapy consisting of 12 g/m2 methotrexate, 60 mg/m2 cisplatin, 75 mg/m2 adriamycin and 15 g/m2 ifosfamide. In both patients, serum methotrexate levels fell to below 0.2 micromol/l within 48-96 h during initial treatment cycles. However, following recent exposure to therapeutic vancomycin in the preceding 10 days and in the absence of overt renal impairment, both patients manifested markedly prolonged methotrexate clearance, requiring 170 231 h to reach serum levels of less than 0.2 microM. Subclinical renal impairment was documented by impaired glomerular filtration rates in both cases by technetium 99 m diethylene triamine penta-acetic acid scanning. Subsequent methotrexate cycles using an unmodified schedule were cleared within 72 h. Both cases had their glomerular filtration rate re-assessed, which showed marked improvement. CONCLUSIONS: Recent exposure to vancomycin, even in the absence of overt renal impairment, may adversely affect methotrexate excretion, which can subsequently lead to increased toxicity of the antimetabolite. The glomerular filtration rate should be measured in such cases so that appropriate dose modification of methotrexate can be made. PMID- 11886014 TI - Second-line treatment with docetaxel after failure of a platinum-based chemotherapy in squamous-cell head and neck cancer. PMID- 11886015 TI - Testicular sperm extraction prior to treatment in azoospermic patients with Hodgkin's disease. PMID- 11886016 TI - Is 18 months too early for the chat? PMID- 11886017 TI - Genetic and environmental influences on the cognitive outcomes of children with fragile X syndrome. AB - OBJECTIVE: To measure the genetic and environmental factors influencing the cognitive outcomes in children with fragile X, a common genetic disorder causing cognitive impairments. METHOD: In-home evaluations were conducted on 120 children (80 boys and 40 girls) with the fragile X full mutation and their unaffected siblings. RESULTS: Multiple regression analyses show that the cognitive outcomes for girls with fragile X are most strongly predicted by the mean IQ of their parents, with a small proportion of the variance accounted for by the quality of their home environment. FMR1 protein (FMRP) was associated with girls' levels of distractibility. Mean parental IQ was associated only with boys' Performance IQs, while FMRP was associated with boys' Full Scale IQs. The quality of boys' home environments accounted for more of the variance in their cognitive outcomes than it did for affected girls. CONCLUSIONS: Both biological/genetic factors and environmental factors are significant predictors of IQ in children with fragile X syndrome; however, the influence of specific factors differs between girls and boys. These findings lay the foundation for further investigation into biological and environmental interventions. PMID- 11886018 TI - Development, reliability, and validity of the children's aggression scale-parent version. AB - OBJECTIVE: To provide preliminary psychometric data on the Children's Aggression Scale-Parent Version (CAS-P), which assesses severity, frequency, pervasiveness, and diversity of aggressive, as distinct from nonaggressive, disruptive behaviors. METHOD: The scale has 33 items representing five domains: Verbal Aggression, Aggression Against Objects and Animals, Provoked Physical Aggression, Unprovoked Physical Aggression, and Use of Weapons. The CAS-P was completed for 73 clinically referred children. Validity was evaluated dimensionally by examining the relationship of CAS-P scores to other parent and teacher rating scales, and categorically by comparing scores of children with attention-deficit hyperactivity disorder (ADHD) alone, oppositional defiant disorder, and conduct disorder. RESULTS: The scale as a whole had excellent internal consistency (alpha = .93). Children with conduct disorder were rated significantly higher than those with oppositional defiant disorder, who were rated significantly higher than those with ADHD alone. The CAS-P did not distinguish clinical control children from those with ADHD only. Correlations with other rating scales provide further support for the validity of the CAS-P. CONCLUSIONS: The CAS-P assesses distinct components of aggressive behavior and may fill a gap in that it distinguishes among various types and severity of aggressive behaviors, and the settings in which they take place. PMID- 11886019 TI - Psychopharmacology and aggression. I: A meta-analysis of stimulant effects on overt/covert aggression-related behaviors in ADHD. AB - OBJECTIVE: To determine by meta-analysis the effect size for stimulants on overt and covert aggression-related behaviors in children with attention deficit/hyperactivity disorder (ADHD), separately from stimulant effects on the core symptoms of ADHD. METHOD: A review of the literature from 1970 to 2001 revealed 28 studies meeting inclusion/exclusion criteria for meta-analysis. These studies yielded 28 independent effects of overt aggression and 7 independent effects of covert aggression. RESULTS: The overall weighted mean effect size was 0.84 for overt and 0.69 for covert aggression related behaviors in ADHD. Comorbid conduct disorder is associated with diminishing stimulant effect size for overt aggression. CONCLUSION: Stimulant effects for aggression-related behaviors in ADHD have effect sizes similar to those for the core symptoms of ADHD. PMID- 11886020 TI - Psychiatric comorbidity and functioning in clinically referred preschool children and school-age youths with ADHD. AB - OBJECTIVE: Although the literature documents that attention-deficit/hyperactivity disorder (ADHD) commonly onsets prior to age 6, little is known about the disorder in preschool children. We evaluated the clinical characteristics, psychiatric comorbidity, and functioning of preschool children and school-age youths with ADHD referred to a pediatric psychiatric clinic for evaluation. METHOD: Structured psychiatric interviews assessing lifetime psychopathology by DSM-III-R criteria were completed with parents about their children. Family, social, and overall functioning were also assessed at intake. RESULTS: We identified 165 children with ADHD aged 4 to 6 years (preschool children) and 381 youths aged 7 to 9 years (school-age) with ADHD. Despite being younger, preschool children had similar rates of comorbid psychopathology compared with school-age youths with ADHD. There was an earlier onset of ADHD and co-occurring psychopathology in the preschool children compared to school-age youths. Both preschool children and school-age youths had substantial impairment in school, social, and overall functioning. CONCLUSIONS: The results of this study suggest that despite being significantly younger, clinically referred preschool children with ADHD are reminiscent of school-age youths with ADHD in the quality of ADHD, high rates of comorbid psychopathology, and impaired functioning. Follow-up of these clinically referred preschool children with ADHD to evaluate the stability of their diagnoses, treatment response, and their long-term outcome are necessary. PMID- 11886022 TI - Children's responses to low parental mood. I: Balancing between active empathy, overinvolvement, indifference, and avoidance. AB - OBJECTIVE: The aims were to study children's behavioral and emotional responses to parental low mood, and the associations of these responses with parental depressive symptoms and the children's own mental distress. METHOD: The community sample consists of 990 twelve-year-old Finnish children and their mothers (843) and fathers (573). The children's responses were elicited by inquiring about their behaviors and feelings when their mothers and their fathers were feeling down. RESULTS: Cluster analysis identified four response patterns: the Active Empathy, Emotional Overinvolvement, Indifference and Avoidance. The Emotional Overinvolvement and the Avoidance groups reported more depressive and externalizing symptoms than the other two groups. The patterns did not vary on the basis of variation in the level of parental depressive symptoms. CONCLUSIONS: Children are sensitive members of their families and feel for their parents. Their responses can reflect both adaptive and nonadaptive patterns. PMID- 11886021 TI - Socioeconomic status as a moderator of ADHD treatment outcomes. AB - OBJECTIVE: To explore whether socioeconomic status (SES) variables moderate treatment response of attention-deficit/hyperactivity disorder (ADHD) to medication management (MedMgt), behavioral treatment (Beh), combined intervention (Comb), and routine community care (CC). METHOD: The MTA Cooperative Group's intent-to-treat (ITT) analyses were repeated, covarying for composite Hollingshead SES, education, occupation, income, and marital status. RESULTS: Individual SES variables were more informative than the composite Hollingshead Index. Treatment response of children from less educated households paralleled ITT outcomes: no significant difference was found between Comb and MedMgt (both better than Beh and CC) for core ADHD symptoms. However, children from more educated families showed superior reduction of ADHD symptoms with Comb. For oppositional-aggressive symptoms, children from blue-collar, lower SES households benefited most from Comb, whereas those from white-collar, higher SES homes generally showed no differential treatment response. Household income and marital status failed to influence outcomes. Controlling for treatment attendance attenuated the moderating effects of the SES variables only for MedMgt. CONCLUSIONS: Investigators are encouraged to use independent SES variables for maximal explanation of SES effects. Clinicians should prioritize target symptoms and consider the mediating role of treatment adherence when determining an ADHD patient's optimal intervention plan. PMID- 11886023 TI - Children's responses to low parental mood. II: Associations with family perceptions of parenting styles and child distress. AB - OBJECTIVE: In an earlier article (part 1) the authors identified four patterns of children's responses to parental low mood: Active Empathy, Emotional Overinvolvement, Indifference, and Avoidance. They then hypothesized that these response patterns were related to parenting styles and to discrepancies in family members' perceptions of parenting and child mental distress. METHOD: A normal population sample of 990 twelve-year-old Finnish children and their mothers (843) and fathers (573) was used. Within-family multivariate analyses conducted in mother-father-child triads (470) were used to examine whether quality of parenting varied according to children's responses and whether parents' and children's perceptions of parenting and child distress were different. RESULTS: Children in the Active Empathy and Indifference groups experienced more positive parenting than those in the other two groups. Discrepancies in family members' perceptions of child distress and mothering and fathering were especially characteristic of the Emotional Overinvolvement group. Typical for the Avoidance group was a within-family agreement on poor parenting and severe child distress. CONCLUSIONS: Children's response patterns as regards parental low mood are related to family dynamics. The study suggests that discrepancies in parents' and children's perceptions of parenting and child distress can be meaningful in understanding family interactions and child development and well-being. PMID- 11886024 TI - Parental identification of depression and mental health service use among depressed adolescents. AB - OBJECTIVE: To examine a proposed pathway to service use among depressed adolescents, this study assessed the effects of (1) parental perceptions of family burden due to adolescents'depression, (2) adolescent-parent communication, (3) parents' depressive symptomatology, and (4) comorbid substance use disorders on parental identification of adolescent depression and use of mental health services. METHOD: A two-stage screening process was used to recruit 44 depressed adolescents and their parents from pediatrics clinics between 1997 and 1999. Measures included structured diagnostic interviews with adolescents, the Child and Adolescent Services Assessment, the Child and Adolescent Functional Assessment Scale, the Beck Depression Inventory (for parents), and the Communication subscale of the Inventory of Parent and Peer Attachment. RESULTS: When history of service use was controlled, two predictors--parental perceptions of family burden and presence of a substance use disorder in the adolescent--were most strongly related to parental depression-identification. The data support the role of parental identification of depression as a mediator between the parent/adolescent characteristics and reports of mental health service use. CONCLUSION: Enhancing parents' abilities to identify and understand signs of depression may facilitate service use among depressed adolescents. PMID- 11886025 TI - Group cognitive-behavioral treatment for depressed adolescent offspring of depressed parents in a health maintenance organization. AB - OBJECTIVE: A randomized, controlled effectiveness trial of group cognitive behavioral therapy (CBT) for depressed adolescent offspring of depressed parents in a health maintenance organization (HMO) was conducted. METHOD: Potential adult cases were found by reviewing antidepressant medication prescriptions, mental health appointments, and medical charts. Introductory study letters signed by each parent's treating physician were mailed to the appropriate adults. Eligible offspring aged 13 to 18 who met current DSM-III-R criteria for major depression and/or dysthymia were randomly assigned to either usual HMO care (n = 47) or usual care plus a 16-session group CBT program (n = 41). Assessments were conducted at baseline, after treatment, and at 12- and 24-month follow-up. RESULTS: Using intent-to-treat analyses, the authors were unable to detect any significant advantage of the CBT program over usual care, either for depression diagnoses, continuous depression measures, nonaffective measures, or functioning outcomes. CONCLUSIONS: Group CBT does not appear to be incrementally beneficial for depressed offspring of depressed parents who are receiving other mental health care. However, given that many other studies have found positive effects of CBT for youth depression, this single study should not be viewed as evidence that CBT is ineffective overall. PMID- 11886026 TI - The voice DISC-IV with incarcerated male youths: prevalence of disorder. AB - OBJECTIVES: (1) To accurately assess rate of psychiatric disorder in incarcerated juveniles, and (2) to examine the feasibility of using a self-administered, comprehensive structured psychiatric assessment with those youths. METHOD: In 1999-2000, 292 recently admitted males in secure placement with New Jersey and Illinois juvenile justice authorities provided self-assessments by means of the Voice Diagnostic Interview Schedule for Children-IV, a comprehensive, computerized diagnostic instrument that presents questions via headphones. RESULTS: Assessments were well tolerated by youths, staff, and parents; 92% of approached youths agreed. Rates of disorder were comparable to prior diagnostic assessment studies with interviewers. Beyond expectable high rates of disruptive and substance use disorders, youths reported high levels of anxiety and mood disorders, with over 3% reporting a past-month suicide attempt. Youths with substance use disorder were significantly more likely to be incarcerated for substance offenses than were youths with no disorder or those with other, non substance use disorders. CONCLUSIONS: Although the study identified rates of disorder generally comparable to those of prior investigations, some differences, understandable in the context of measurement variations, are apparent. Those variations offer recommendations for mental health assessment practices for youths in the justice system that would include using a comprehensive self-report instrument, pooling across parent and youth informants for certain disorders, focusing on current disorder, and flexibility regarding consideration of impairment. PMID- 11886027 TI - Violence exposure, posttraumatic stress, and personality in juvenile delinquents. AB - OBJECTIVE: To assess posttraumatic stress and its relationship to comorbid psychopathology, violence exposure, and personality traits in Russian male juvenile delinquents. METHOD: Posttraumatic stress and comorbid psychopathology were assessed by a semistructured psychiatric interview (Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime Version) in 370 delinquent youths during winter-spring of 1999. In addition, violence exposure, personality, and psychopathology were assessed by self-reports. RESULTS: Most delinquents reported some degree of posttraumatic stress: 156 subjects (42%) fulfilled partial criteria and 87 (25%) fulfilled full DSM-IV criteria for posttraumatic stress disorder (PTSD). Violence-related experiences (witnessing and victimization) were the most common types of trauma. Higher levels of posttraumatic stress were accompanied by higher rates of comorbid psychopathology, with the most striking differences occurring between the groups with full versus partial PTSD criteria. Violence exposure was related to temperamental behavior activation (novelty seeking), whereas PTSD symptom scores were predominantly related to behavior inhibition and poor coping (high harm avoidance and low self-directedness). CONCLUSIONS: Similar to findings from American samples, Russian juvenile delinquents represent a severely traumatized population, mainly due to high levels of violence exposure. Those with full PTSD are the most severely traumatized and have highest rates of psychopathology, as compared to those with no or partial PTSD, and they require the most clinical attention and rehabilitation. Both exposure to violence and levels of posttraumatic stress are related to personality traits, which influence degree of exposure and individual perception of stress. The latter should be considered in individualized approaches to rehabilitation. PMID- 11886028 TI - Risperidone versus clonidine in the treatment of children and adolescents with Tourette's syndrome. AB - OBJECTIVE: To evaluate the efficacy and tolerability of risperidone in comparison with clonidine in the treatment of children and adolescents with Tourette's syndrome (TS). METHOD: Following a 7- to 14-day single-blind, placebo lead-in, 21 subjects aged 7 to 17 years were randomly assigned to 8 weeks of double-blind treatment with clonidine or risperidone. Research scales evaluated tics and comorbid obsessive-compulsive and attention-deficit/hyperactivity symptoms. RESULTS: Risperidone and clonidine appeared equally effective in the treatment of tics in an intent-to-treat analysis, as rated by the Yale Global Tic Severity Scale (YGTSS). Risperidone produced a mean reduction in the YGTSS of 21%; clonidine produced a 26% reduction. Among subjects with comorbid obsessive compulsive symptoms, 63% of the risperidone group and 33% of the clonidine group responded to treatment (not significant). The most common adverse event seen with both treatments was sedation, which was mild to moderate in severity. Sedation subsequently resolved with continued administration of the medication or with a dose reduction. No clinically significant extrapyramidal symptoms were observed. CONCLUSIONS: In this pilot study, risperidone demonstrated efficacy equivalent to clonidine in the treatment of tic symptoms in children and adolescents with TS. Further research is needed to clarify the role of atypical antipsychotics in TS and to delineate potential benefits for comorbid obsessive-compulsive and attention-deficit/hyperactivity symptoms. PMID- 11886029 TI - Weight gain associated with olanzapine and risperidone in adolescent patients: a comparative prospective study. AB - OBJECTIVE: To evaluate weight gain associated with olanzapine, risperidone, and haloperidol treatment and its clinical risk factors in adolescent patients. METHOD: The study was conducted at three adolescent psychiatric departments in two mental health centers in the Tel Aviv area. All patients were Jewish Israelis. Weight and body mass index (BMI) of hospitalized adolescents treated with olanzapine (n = 21), risperidone (n = 21), or haloperidol (n = 8) were prospectively monitored on a weekly basis for the first 12 weeks of treatment. Various clinical risk factors were tested for association with weight gain. RESULTS: The olanzapine and risperidone groups experienced significant weight gain between baseline and endpoint (p < .01), whereas the average weight of the haloperidol group did not change. Average weight gain was significantly higher for the olanzapine group (7.2 +/- 6.3 kg, 11.1% +/- 7.8%) than for the risperidone (3.9 +/- 4.8 kg, 6.6% +/- 8.6%) and haloperidol (1.1 +/- 3.3 kg, 1.5% +/- 6.0%) groups. Extreme weight gain (>7%) was recorded in 19 patients (90.5%), 9 patients (42.9%), and 1 (12.5%) patient, respectively Gender (males), low concern about gaining weight (females), low baseline BMI, and paternal BMI were positively correlated with weight gain, whereas previous neuroleptic history, neuroleptic dosage, response to treatment, and illness duration were not. CONCLUSIONS: Olanzapine and risperidone are associated with extreme weight gain in adolescents, much higher than that reported in adults. This side effect should be taken into consideration before prescribing these medications, especially in patients at high risk. PMID- 11886030 TI - Clinical problem solving: the case of Matthew, part III. PMID- 11886031 TI - Genetics of childhood disorders: XXXVI. Stem cell research, part 1: New neurons in the adult brain. PMID- 11886032 TI - Managing breakthrough pain. PMID- 11886034 TI - Ketamine and problems with advanced palliative care in the community setting. PMID- 11886035 TI - Worldwide hospice & palliative care: focus on east Asia. PMID- 11886033 TI - Dancing with angels. PMID- 11886036 TI - Quality end-of-life care: the right of every Canadian. PMID- 11886037 TI - The therapeutic triad. PMID- 11886038 TI - Palliative medicine in a sole community provider. PMID- 11886039 TI - The development of a hospice junior volunteer program. AB - In the United States, volunteer services are mandated by hospice Medicare guidelines; volunteers provide a very valuable service to patients, families, and other members of the interdisciplinary team. A hospice junior volunteer program can engage teens in the care of the dying in our communities. This article describes the development and implementation of a junior volunteer program at St. Thomas Hospice in Hinsdale, Illinois. PMID- 11886040 TI - Nebulized hydromorphone for dyspnea in hospice care of advanced cancer. AB - This case report describes the use of nebulized hydromorphone for management of dyspnea in advanced cancer in home hospice care. The patient was intolerant of morphine; nebulized hydromorphone was used as an alternative to nebulized morphine for dyspnea and found to be both safe and effective. PMID- 11886041 TI - Methylphenidate for depression in hospice practice: a case series. AB - Psychostimulants such as methylphenidate have been used for depression in cancer patients. We report the successful use of methylphenidate to treat depression in 10 consecutive patients with advanced cancer. A rapid onset of effect was noted. Appetite, concentration, fatigue, and sedation also improved in some persons. No severe side effects were noted. PMID- 11886042 TI - Attitudes, values, beliefs, and practices surrounding end-of-life care in selected Kansas communities. AB - OBJECTIVE: This study was designed to better meet end-of-life care needs for patients and their families in rural Kansas communities. METHODS: Initially, statistical information indicated an underutilization of hospice services in rural communities. To evaluate the data, focus groups were conducted in targeted communities in order to determine attitudes, values, beliefs, and practices surrounding end-of-life care. A script was developed using established focus group protocols. Each focus group was recorded and the tapes were transcribed. Transcripts were reviewed and categorized for similarities and emerging key issues. RESULTS: Five major areas of concern were identified using this methodology. Major concerns were: (1) participants believed that control over decisions about care at the end of life is the patient's right; (2) while participants saw a need for advance directives, they avoided using them; (3) group participants believed that the patient's wishes should be given first priority and this was viewed as a patient's right; (4) there was an expressed lack of trust in the existing health care system and its providers; and (5) participants expressed more fear over the manner of death than death itself. They fear a technological death as opposed to a good death. CONCLUSIONS: The values most important to the group participants included: freedom and independence, trust, honesty, the right to information, and the importance of family. This information will be utilized in the development of programs and interventions to effect changes in end-of-life care, not only in Kansas, but also in areas with a comparable population. PMID- 11886043 TI - Palliative care and hospice opioid dosing guidelines with breakthrough pain (BP) doses. PMID- 11886044 TI - The ballet of baseball: lessons of the game for hospice. AB - As Yogi Berra once said, "The future ain't what it used to be." In the era of rapid change in health care, hospice and palliative care programs will survive only through organizational teamwork. Using the lessons of baseball, we present a stadium-eye perspective on how programs can take the three fundamentals of baseball--pitching, batting, and fielding--and translate them into the three fundamentals of organizational teamwork--clinical, operational, and financial. The best clinicians (pitchers) are of no use if the office operations (batting) keep the patients (fans) out of the stadium and no program can survive without the financial resources (fielding.) When you come to a fork in the road, take it. PMID- 11886045 TI - Hope and Hanukkah. PMID- 11886046 TI - Parenteral opioids and cost. PMID- 11886047 TI - Palliative care for children: is it really needed? PMID- 11886048 TI - Compelling statement. PMID- 11886049 TI - Choosing, not withholding. PMID- 11886050 TI - Life and afterlife in Jewish tradition. PMID- 11886051 TI - Out of context? PMID- 11886052 TI - Worldwide hospice & palliative care: focus on Africa. PMID- 11886053 TI - The Netherlands moves toward legalization of assisted suicide and euthanasia. PMID- 11886054 TI - Unique aspects of caring for dying children and their families. AB - Pediatric hospice has become an important service for children and their families in the past decade. In this article we present unique aspects of StarShine, the hospice of Children's Hospital Medical Center in Cincinnati, Ohio. Length-of service data demonstrate the need to find innovative ways of encouraging earlier referral to hospice. Several specialized aspects of care, such as long-term bereavement, pet visitation and social services for the family are presented. The initiation of a novel in-home pain management program is described. StarShine has not followed an adult hospice model, and as such, we discuss those unique aspects of dying pediatric patients and suggest specific solutions and interventions designed for children. PMID- 11886055 TI - Turning toward death together: conversation in mortal time. AB - Death has a different meaning for everyone it touches, and these meanings have serious impact on how each person communicates with others as they deal with it together. This paper is a reflection on clinical experience in dealing with the dying and their caregivers in a comprehensive cancer center. By "mortal time," we mean the psychological state human beings enter when confronted with the prospect of death. Our focus is on a particular and powerful instance of the entry into mortal time--the diagnosis of a terminal illness. The experience of mortal time is profoundly subjective. Authentic conversation has the power not only to enhance how people cope practically with dying, but to illuminate and enrich the very meaning of life for patients and caregivers alike, as they enter the sacred moment of mortal time together. PMID- 11886056 TI - The relationship of pain and suffering in a hospice population. AB - Although suffering is frequently encountered in the hospice setting, few studies examine this condition. The purpose of this study was to examine the relationship between terminally ill hospice patients' pain and their physical, spiritual, and personal or family suffering. Using a tool developed to measure suffering in those categories, a convenience sample of 92 patients were asked to rate their worst pain within the last 24 hours, and to rate their suffering at the time of the interview. All items were rated on a 0-10 Numeric Intensity Scale. Pain scores and suffering scores were divided into four categories; no pain or no suffering (0), mild pain or mild suffering (1-3), moderate pain or moderate suffering (4-6), and severe pain or severe suffering (7-10). Mean scores were compared for pain and suffering. More patients experienced suffering than pain. The highest mean suffering scores occurred in the severe pain category. Correlation coefficients for each suffering and pain category were also calculated. Results indicated a statistically significant correlation only between severe pain and suffering in the categories of loss of enjoyment of life, unfinished business, and concern for loved ones. Data indicated that patients view pain and suffering as separate entities. Further research is needed to better define the relationship between pain and suffering in order to improve assessment and intervention in a hospice setting. PMID- 11886057 TI - Self-esteem in a palliative care population. AB - OBJECTIVE: To study the effects of palliative care treatment in cancer patients, on their self-esteem. DESIGN AND SETTING: It was a cross-sectional study conducted at Shaukat Khanum Memorial Cancer Hospital and Research Center, Lahore, Pakistan (SKMCH & RC). PARTICIPANTS: A total of 60 referred cancer patients receiving palliative care were interviewed using the Mehrabian self-esteem scale (MSE). No follow-up was required. All the patients were over 16 years of age. The sample was not restricted to any one type of cancer. Since lack of education could be a contributing factor, only patients who had a minimum of an eighth grade education were included in the study. RESULTS: Four variables were studied: gender, age, diagnostic-related group, and performance status. Performance status was determined using the Eastern Cooperative Oncology Group Scale (ECOG). The only significant correlation was between self-esteem and performance status, with a p value of 0.04. CONCLUSION: We conclude that the patients who are functionally active (not disabled) have a higher level of self-esteem, as compared to those who are disabled. This is quite understandable, as the patients who are not dependent on others for their needs feel worthy, resulting in higher self-esteem. This was the first study of its kind done in Pakistan. We identified two limitations. First, the sample size was small. Second, although the Mehrabian self-esteem scale is a valid, reliable, and standardized scale, it was not standardized for the population of Pakistan. PMID- 11886058 TI - Relieving the agony of the new pain management standards. AB - The Joint Commission on Accreditation of Healthcare Organizations (JCAHO) has issued new standards for pain assessment in accredited hospitals and other health care settings, including hospice and home care. Under the new pain management standards, health care facilities will be called upon to recognize the right of patients to appropriate assessment and management of pain; to assess the existence of pain, its nature, and intensity; to record the results of the assessment in a way that facilitates regular reassessment and follow-up; to determine and ensure staff competency in pain assessment and management, and to address pain assessment and management in the orientation of all new staff; to establish policies and procedures that support the appropriate prescription or ordering of effective pain medications; to educate patients and their families about effective pain management; and to address patient needs for symptom management in the discharge planning process. Many health care organizations are reporting confusion and lack of understanding about the scope of the new standards. To address this issue, this article summarizes the new pain management standards. This article is based on a three-part series published in the Journal of Healthcare Safety, Compliance & Infection Control (January, March, and April 2000). PMID- 11886059 TI - Hydrocodone for cough in advanced cancer. AB - Cough is a common symptom in advanced cancer. Hydrocodone is the antitussive of choice in our palliative medicine inpatient unit. We reviewed the pharmacy records for the use of hydrocodone for all cancer admissions to our unit from May 1996 to December 1998. Median treatment duration with hydrocodone was three days (range 1-18). Median maximum daily dose was 15 mg (range 5-100), and median total dose during the hospital stay was 32 mg (range 5-455). Lung cancer as a primary cancer site was strongly related to the use of hydrocodone. The highest median duration of treatment (five days) was for esophageal cancer and the highest median maximum daily dose (35 mg) and total dose (75 mg) were for treating kidney cancer. This retrospective review provides information regarding the use of hydrocodone on the palliative medicine unit of the Cleveland Clinic Foundation. Controlled trials are needed to evaluate the efficacy and safety of hydrocodone for cough in advanced cancer. PMID- 11886061 TI - The transitional phase: the closing journey for patients and family caregivers. AB - The ability to identify physical waypoints along the continuum of illness may give end-stage patients and their families opportunities for goal reframing and risk reduction. That period which exists between active participation in the activities of daily living and a bedbound status, herein described as the transitional phase, is characterized by the four precursive, or seminal, behaviors of anorexia, increased sleep, weakness, and confusion, and the two cardinal behaviors of incontinence and falls. It is a time of heightened anxiety for families and risk of injury for patients. The ability to identify these behaviors as part of a definable phase, with a beginning and an end, allows the health care clinician or hospice worker to educate the family, assisting with goal reassignment, risk reduction, and diminishment of anxiety. Family understanding of the finite nature of the transitional phase may also reduce the need for placement outside the home for those wishing their loved one to die at home, and provide the caregiver with meaningful participation in end-of-life problem solving. PMID- 11886060 TI - Use of continuous ambulatory infusions of concentrated subcutaneous (s.q.) hydromorphone versus intravenous (i.v.) morphine: cost implications for palliative care. AB - Health care practitioners are increasingly under pressure to curtail spending while trying to deliver excellent patient care. These issues are also affecting palliative care, particularly now that palliative care programs are expanding. A comparison of cost-effectiveness and feasibility of using continuous subcutaneous (s.q.) ambulatory infusion of hydromorphone versus intravenous (i.v.) ambulatory morphine is illustrated in this study. With the high doses of morphine required in chronic cancer pain, the use of subcutaneous morphine is not feasible due to the volume of solution required to be delivered. Hydromorphone can be prepared in concentrated solutions enabling it to be delivered by the subcutaneous route. Morphine stability data are available. However, hydromorphone stability has only been verified for seven days; thus, stability data were needed post-seven days. Concentrations of 10 mg/ml, 20 mg/ml, 50 mg/ml, and 100 mg/ml, in 0.9 percent normal saline or dextrose 5 percent water, were analyzed via high-performance liquid chromatography (HPLC) at seven and 28 days. Cost comparisons of supplies and associated costs with subcutaneous versus intravenous solutions were obtained. Hydromorphone was found to be stable for 28 days in both dilutants. Cost analysis of a hydromorphone 28-day supply resulted in substantial savings over the equivalent costs of morphine infusions. PMID- 11886062 TI - Terminal sedation. PMID- 11886063 TI - Nursing responds to the demand for improved end-of-life care. PMID- 11886064 TI - Politics, cancer pain, and methadone. PMID- 11886066 TI - The expertise of hospice professionals leads to quality end-of-life care. PMID- 11886065 TI - What's in a name? NHO becomes NHPCO and the end-of-life debate goes mainstream. PMID- 11886067 TI - Supportive/palliative care of children suffering from life-threatening and terminal illness. PMID- 11886068 TI - A comparison of hospice in the UK and the US. AB - This article presents a comparison of hospice services in the United Kingdom and the United States. The article includes a brief history of the development of the modern hospice movement in the United States and the United Kingdom, with emphasis on funding, regulatory standards, and demographics. PMID- 11886069 TI - Who are our clients? A profile of a community-based Buddhist hospice. AB - One of the ongoing challenges for those involved in the hospice movement is to find ways of effectively extending the provision of hospice services to all in need. Pragmatic, empirical research that provides information on the present and potential hospice population is essential to foster the development of services if this challenge is to be met. This discussion presents a descriptive profile of the clients of a community-based Buddhist hospice in Brisbane, Australia, known as the Karuna Hospice Service (KHS). This descriptive profile makes a contribution to the important task of establishing where we currently are, in the hope that this will begin to delineate where fresh energy needs to be directed in the future. PMID- 11886070 TI - The Jewish patient and terminal dehydration: a hospice ethical dilemma. AB - Culturally competent nursing care regarding the ethical dilemma of terminal dehydration (withholding or withdrawing food and fluid) for the Jewish hospice patient involves applying the ethical principles of justice, autonomy, beneficence, and nonmaleficence to nursing interventions by identifying outcomes that focus on the high value Jews place on life; avoiding stereotyping as to what it means to be Jewish; knowledge of various Jewish traditions surrounding death and dying; and good communication with the patient and his or her family. PMID- 11886071 TI - Terminal sedation for existential distress. AB - Although sedation for existential distress has been actively discussed in the palliative care literature, empirical reports are limited. A retrospective cohort study was performed to clarify the physical conditions of terminally ill cancer patients who expressed existential distress and received sedation. Of 248 consecutive hospice inpatients, 20 patients expressed a belief that their lives were meaningless and received sedation. The target symptoms for sedation were dyspnea (n = 10), agitated delirium (n = 8), and pain (n = 1). Only one patient received sedation for psychological distress alone, although physical symptoms were acceptably relieved. The Palliative Performance Scale just before sedation was 10 (n = 7), 20 (n = 11), 30(n = 1), and 40(n = 1). All but one patient could take nourishment orally of only mouthfuls or less. Edema, dyspnea at rest, and delirium were observed in 10, 13, and 14 cases, respectively. The Palliative Prognostic Index was greater than 6.0 in all but one case with a mean of 12 +/- 3.3. In conclusion, in our practice, sedation was principally performed for physical symptoms of cancer patients in very late stages. Further research is encouraged to establish standard therapy for existential distress of the terminally ill. PMID- 11886072 TI - Clinical applications of methadone. PMID- 11886074 TI - A unique group of self-splicing introns in bacteriophage T4. AB - We describe in this review, the salient splicing features of group I introns of bacteriophage T4 and propose, a hypothetical model to fit in the self-splicing of nrdB intron of T4 phage. Occurrence of non-coding sequences in prokaryotic cells is a rare event while it is common in eukaryotic cells, especially the higher eukaryotes. Therefore, T4 bacteriophage can serve as a good model system to study the evolutionary aspects of splicing of introns. Three genes of T4 phage were found to have stretches of non-coding sequences which belonged to the group IA type introns of self-splicing nature. PMID- 11886073 TI - Control theory in dying: what do we know? AB - As we try to develop appropriate models for end-of-life care, it is important to review theoretical applications as they relate to current models of care. It is also vital to listen carefully to what patients and theirfamily caregivers tell us about their needs at the end of life. This article explores the issue of control: its application to wellness at the end of life, and patient and family caregiver perceptions about end-of-life control. Interventions and implications for professional caregivers are proposed. PMID- 11886076 TI - Mercury induced modifications in the stereochemistry of the active site through Cys-73 in a serine protease--crystal structure of the complex of a partially modified proteinase K with mercury at 1.8 A resolution. AB - Proteinese K (PK) isolated from Tritirachium album Limber was crystallized with HgCl2 in excess, under microgravity conditions. The intensity data were collected at 4 degrees C to 1.8 A resolution and the final R-factor after refinement for all the reflections was 0.164. Mercury has been found at two sites with partial occupancies (0.4 and 0.6) which are at distances of 2.48 A and 2.58 A respectively from Cys-73 Sgamma. The Cys-73 in the enzyme structure is located close to the active site residue, His-69. This region is completely buried and is not accessible to the solvent. It is rather tightly packed. Therefore, the binding of mercury distorts the stereochemistry of the neighbouring residues including those belonging to the catalytic triad. As a result of this, the Ogamma of Ser-224 is displaced by 0.6 A which causes the inactivation of proteinase K by increasing the H-bond distance to 3.7 A between Ser-224 Ogamma and His-69 Nepsilon2. PMID- 11886077 TI - An alternative approach for screening active bam HI variants: overexpression in T 7 RNA polymerase based system. AB - The type II restriction endonuclease, Bam HI, has been overexpressed in E. coli by cloning the Bam HI gene in frame with an E. coli Ribosome Binding Site (RBS) under the T7 promoter of an E. coli expression vector pRSET A. The expression level of Bam HI endonuclease using this construct was found to be higher than that reported of the overexpressing clone pAEK14. Our overexpressing clone, pAABRw in BL21 cells in presence of Bam HI methylase in pMAP6 following induction with IPTG yields about 9.2 x 10(6) units per gram wet cell paste. In vivo activity of the recombinant endonuclease could be confirmed by the SOS induction assay in JH139 cells even in the absence of T7 polymerase and cognate Bam HI methylase because of leaky expression in E. coli. This provides an alternate way to screen the active endonuclease and its variants. PMID- 11886075 TI - Molecular modelling of epitope presentation using membrane protein OmpC. AB - Three-dimensional models of the chimeric S. typhi OmpC protein carrying an epitope from rotavirus VP4 capsid protein on either of two exposed loops (fourth and sixth) were constructed separately, using computer-aided homology modelling. The theoretical model of S. typhi OmpC was used as a template. The monomers were initially energy minimized. The trimers were generated for both the chimeric S. typhi OmpC proteins and the structures were optimized after several cycles of minimization. The surface accessibility calculations for the resulting models show that epitope recognition should be more effective in the fourth loop than in the sixth loop, in accordance with the experimental results on the immunogenic nature of the rotaviral epitope inserted into the two putative loops of S. typhi OmpC. PMID- 11886078 TI - Role of positive charge of lysine residue on ribosome-inactivating property of gelonin. AB - The report that gelonin cross-linked with monoclonal antibodies with the use of 2 iminothiolane (2-IT) exhibited higher cytotoxicity than the conjugates prepared with the use of N-succinimidyl-3-(2-pyridylthio) propionate (SPDP) alone, has prompted us to investigate the effect of epsilon-NH2 group modification with 2-IT on the ribosome-inactivating property (RIP) of gelonin. The purified gelonin was modified with 2-IT at a different molar ratio and their effects on immunoreactivity and ribosome-inactivating property were compared with those of N succinimidyl 6-[3-(2-pyridyldithio) propionamido] hexanoate (long chain-SPDP) and SPDP modified gelonin derivatives. Modification of single amino group with 2-IT results in about 25-50% inhibition of immunoreactivity and 60-70% loss of protein synthesis inhibition activity. Modification of 2-3 amino groups further hampers both immunoreactivity and protein synthesis inhibition property of gelonin. Both the long chain-SPDP with SPDP modifications showed more pronounced effects on immunoreactivity and RIP activity as compared to the similar ratio of 2-IT modification(s). It may, therefore, be concluded that the positive charge plays an important role in the immunological as well as the protein synthesis inhibitory effect of gelonin. PMID- 11886079 TI - Binding of globular proteins to DNA from surface tension measurement. AB - Extent of binding (gammap) of globular proteins to calf-thymus DNA have been measured in mole per mole of nucleotide as function of equilibrium protein concentration. We have exploited measurement of the surface tension of the protein solution in the presence and absence of DNA to calculate the binding ration (gammap). Interaction of bovine serum albumin with DNA has been studied at different pH. Interaction of bovine serum albumin with DNA has been studied at different pH, ionic strength and in presence of Ca2+. Interaction of BSA with denatured DNA has also been investigated. Binding isotherms for other globular proteins like beta-lactoglobulin, alpha-lactalbumin and lysozyme have been compared under identical physicochemical condition. It has been noted with considerable interest that globular form of protein is important to some extent in protein-DNA interaction. An attempt has been made to explain the significance of difference in binding ratios of these two biopolymers in aqueous medium for different systems in the light of electrostatic and hydrophobic effects. Values of maximum binding ration (gammap(m)) at saturated level for different systems have been also presented. The Gibb's free energy decrease (-deltaG0) of the binding of proteins to DNA has been compared more precisely for the saturation of binding sites in the DNA with the change of activity of protein in solution from zero to unity in the rational mole fraction scale. PMID- 11886080 TI - Age-dependent variations in mitochondrial and cytosolic antioxidant enzymes and lipid peroxidation in different regions of central nervous system of guinea pigs. AB - The age-related changes in the activities of antioxidant enzymes of mitochondrial and cytosolic fractions were measured in different regions of the central nervous system (CNS) in 10 and 32 months old guinea pigs. In old animals, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were reduced (p < 0.05) in all the regions of CNS studied but catalase (CAT) declined significantly only in the cerebral cortex, hypothalamus and cerebellum. Glutathione reductase (GRd) activity declined in cerebral cortex and hypothalamus in the cytosolic fractions and only in cerebellum in the mitochondrial fraction. It is concluded that age-related decline in the activities of antioxidant enzymes is both region and enzyme specific. The endogenous lipid peroxide was found to be significantly higher (p < 0.05) in the 32 month old animals whereas, lipid peroxidation after incubating the tissue homogenate in air was found to be lower (p < 0.05). The in vitro mitochondrial lipid peroxidation decreased with age. The results indicate that accumulation of lipid peroxides takes place with ageing but the susceptibility of lipid peroxidation decreases in the older animals. PMID- 11886081 TI - Polyunsaturated fatty acids (PUFA) regulate neurotransmitter contents in rat brain. AB - The effects of feeding of 6-propylthiouracil (6-PTU) and polyunsaturated fatty acids (PUFA) independently and in combination and administration (ip) of a single dose of triiodothyronine (T3) (2.5 microg/100 g body wt) along with feeding of 6 PTU and PUFA were studied in rat brain. Dopamine (DA), 5-hydroxytryptophan (5 HTP), serotonin (5-HT), 5-hydroxy indole acetic acid (5-HIAA), norepinephrine (NE) and epinephrine (EPI) contents were assayed in the hypothalamus and cerebral cortex regions. It was found that 6-PTU feeding resulted in decrease in dopamine, 5-HT, 5-HTP and 5-HIAA in both regions. In animals fed with PUFA followed by administration of T3, the DA level was found normal. PMID- 11886082 TI - Insulin receptor tyrosine kinase activity in monocytes of type 2 diabetes mellitus patients receiving oral L-lysine. AB - The action of lysine as an antidiabetic agent was examined in human volunteers. Eight patients with type 2 DM were orally supplemented with L-lysine hydrochloride 1 g/day in two doses along with antidiabetic tablets (glyciphage or chlorformine), for a period of two months. Periodically their plasma fasting sugar and insulin receptor tyrosine kinase activity was measured in their monocytes. Eight normal healthy volunteers served as controls for comparison of receptor tyrosine kinase activity. Insulin receptor tyrosine kinase was isolated from monocytes by immunoprecipitation and the activity was determined using exogenous substrate poly glu-tyr (4:1) and radioactive ATP. Phosphorylated peptide was separated by electrophoresis and quantified using a liquid scintillation system. The enzyme activity was significantly low (22074 +/- 1728 dpm/ml immunoprecipitate) in subjects with diabetes when compared to non-diabetic control group (50,775 +/- 3597). Lysine treatment enhanced the enzyme activity by 31% in patients with diabetes and decreased their blood sugar by 27%. PMID- 11886083 TI - Isolation and characterization of NADP+-linked isocitrate dehydrogenase of germinating pea seeds (Pisum sativum). AB - NADP+-linked isocitrate dehydrogenase (E.C.1.1.1.42) has been purified to homogeneity from germinating pea seeds. The enzyme is a tetrameric protein (mol wt, about 146,000) made up of apparently identical monomers (subunit mol wt, about 36,000). Thermal inactivation of purified enzyme at 45 degrees and 50 degrees C shows simple first order kinetics. The enzyme shows optimum activity at pH range 7.5-8. Effect of substrate [S] on enzyme activity at different pH (6.5 8) suggests that the proton behaves formally as an "uncompetitive inhibitor". A basic group of the enzyme (site) is protonated in this pH range in the presence of substrate only, with a pKa equal to 6.78. On successive dialysis against EDTA and phosphate buffer, pH 7.8 at 0 degrees C, yields an enzymatically inactive protein showing kinetics of thermal inactivation identical to the untreated (native) enzyme. Maximum enzyme activity is observed in presence of Mn2+ and Mg2+ ions (3.75 mM). Addition of Zn2+, Cd2+, Co2+ and Ca2+ ions brings about partial recovery. Other metal ions Fe2+, Cu2+ and Ni2+ are ineffective. PMID- 11886084 TI - Purification and properties of antiviral proteins from the leaves of Bougainvillea xbuttiana. AB - A non-phytotoxic, resistance inducing, proteinaceous antiviral principle was purified by ammonium sulphate fractionation, ion exchange chromatography and gel filtration from the leaves of Bougainvillea xbuttiana. It imparted resistance against tobacco mosaic virus (TMV) and sunnhemp rosette virus (SRV) in their respective test hosts viz. Nicotiana glutinosa, N. tabacum var. Samsun NN, and Cyamopsis tetragonoloba, respectively. The purified principle eluted as a single peak upon gel filtration, but exhibited two polypeptides on SDS-PAGE with Mr 28,000 and 24,000. The two polypeptides were found to be highly basic, rich in lysine with pI around 10.0 and 10.5, respectively. Since this principle effected local lesion inhibition in both treated and untreated top leaves of test host, it might be acting in the initial stages of virus infection as a systemic inducer. PMID- 11886085 TI - Lead in a Nigerian savanna soil under long-term cultivation. AB - Concern about heavy metal accumulation in agricultural soils under long-term application of phosphate fertilizers and organic wastes makes investigation of heavy metals in agricultural soils imperative. This study examines the total, available and chemical forms of lead (Pb) in a savanna soil after 50 years of continuous cultivation and application of NPK fertilizers and farmyard manure (FYM). Total Pb concentration ranged from 28 to 42 mg kg(-1) over 2-3 times more than the average Pb concentration of non-polluted soils worldwide. Available Pb was, however, less than 2 mg kg(-1) indicating that Pb in the soils was largely insoluble. There were no detectable concentrations of water soluble and organically-bound Pb fractions. On average, residual Pb accounted for over 80% of total Pb. Compared to the natural site, cultivation and fertilization with NPK and FYM increased total Pb concentration by 19 and 17%, respectively, or, on mass basis, by 10 and 35 kg ha(-1), respectively, after 50 years. Soil Pb showed strong linear relations with sand fraction and inorganic phosphorus in the soils. Thermodynamic equilibrium relations provided some indirect evidence that the control on soluble Pb appeared to be chloropyromorphite [Pb5(PO4)3Cl], an insoluble lead phosphate mineral. PMID- 11886086 TI - Waste ashes for use in agricultural production: II. Contents of minor and trace metals. AB - The present study was carried out to examine the contents of 18 minor and trace metals in five typical municipal waste ashes in Japan. In the waste ashes, Li, Ga, Rb, Y, Zr had relatively higher concentrations, approximately 5-300 mg kg( 1), the remaining metal concentrations were generally approximately 0.05-20 mg kg(-1). A comparison of the metal concentrations in the waste ashes and in Japanese agricultural soils indicated that the ratios for Ga, Mo, Ag, Sb, W, Bi between sewage sludge ash (SSA) and the soils were approximately 10-100 and for the remaining metals approximately 0.2-2; the ratios between food scrap ash (FSA), animal waste ash (AWA), horticulture waste ash (HWA) and incinerator bottom ash (IBA) and the soils were approximately 0.2-5. Furthermore, an overall evaluation on the waste ashes was also carried out using factor analysis with the addition of the other 21 elements examined in a companion paper. In the waste ashes, the major nutrient elements and heavy metals were mainly described by four factors: factors 1 and 2 explained the main information of the minor and trace metals while factors 3 and 4 explained that of the major nutrient elements. Factor 2 in the score plots could be used to evaluate the potential risk of the waste ashes to agricultural soils. Of the five types of waste ashes, SSA and IBA were abundant with minor and trace metals; AWA was relatively abundant with major nutrient elements especially for K; FSA was relatively abundant with major nutrient elements except for K, while HWA was not abundant with either of them. PMID- 11886087 TI - Residuals of beta-hexachlorocyclohexane, dichlorodiphenyltrichloroethane, and hexachlorobenzene in serum, and relations with consumption of dietary components in rural residents in Japan. AB - To estimate levels of organochlorine residuals in the Japanese population and the contribution of dietary factors to these levels, we determined serum levels of beta-hexachlorocyclohexane (beta-HCH), hexachlorobenzene (HCB), p,p' dichlorodiphenyldichloroethane (p,p'-DDD), 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p'-DDE) and p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT) in 41 volunteers (14 men and 27 women) in a rural area of Northern Japan. These organochlorine levels were measured using gas-chromatography mass-spectrometry. By a self-administered dietary history questionnaire, the usual dietary intake was estimated. Their median levels (range) were as follows: beta-HCH, 0.50 (0.05 1.50); HCB, 0.20 (0.02-0.70); and total DDT (p,p'-DDE + p,p'-DDT), 5.0 (0.9-31.0) ng/ml serum. Levels of p,p'-DDD were detected in only seven subjects (0.05-0.6 ng/ml serum). The beta-HCH levels were increased with rice and milk intakes, but the least squares means were not simply increased according to the quartile of the intakes. Concerning HCB, fish intake showed a borderline significant correlation (0.20, P = 0.052). In terms of total DDT, intakes of meat, fish, vegetable and milk showed a positive relationship, although none of them provided statistically significant results. No other statistically significant relation between any organochlorines and any food intakes examined was observed in this study. The present study suggests that organochlorine compounds are transported into the human body via foods in the Japanese population. Their effects on health should thus be investigated and monitored. PMID- 11886088 TI - Trace element levels in whole blood and serum from Swedish adolescents. AB - Blood and serum samples from 372 15-year-old adolescents were collected in two cities in Sweden and analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The objective was to (1) determine the levels of 13 elements in blood and serum from the teenagers; and (2) for each element, investigate the correlation between the concentrations in blood and serum. The concentrations in blood and serum were generally in line with that usually reported for the essential elements Co, Cu, Zn and Se, and generally low for the 'non-essential' elements Cd, Hg, Pb. The median concentrations were in blood and serum, respectively: of Co 0.31 and 0.48 microg/l, Cu 0.92 and 1.0 mg/l, Zn 6.1 and 0.99 mg/l, Se 110 and 100 microg/l, Rb 2.8 and 0.24 mg/l, Hg 1.1 and 0.44 microg/l, Pb 16 and 0.33 microg/l. The median concentration of W in blood was <0.2 microg/l (below the detection limit) and in serum 0.087 microg/l. The median concentrations of Cd, Rh, Pd, Pt and Tl were below the detection limits. Statistically significant correlations were found between the concentrations in blood and serum for Co, Cu, Zn, Se, Rb, W, Hg and Pb. The levels presented in this study constitute baseline levels or levels generally not exceeded in adolescents for 13 elements, including essential, ubiquitous toxic, and rare elements. PMID- 11886089 TI - Fate of POPs (DDX, HCHs, PCBs) in upper soil layers of pine forests. AB - Organochlorines (HCH isomers, DDX and individual PCBs) were determined in pine needle litter and upper soil layers of three forest test areas in central Germany. The contents accumulating over a number of years or even decades in the organic surface layer are compared with the levels of new inputs from needle fall as well as with the levels of older inputs in the upper mineral soil layer. Differences in behaviour between the soil horizons are discussed, especially concerning the DDX and HCH groups. With approximately comparable Corg values (approx. 21-24%) the pH value in the range of 4.24-2.90 in the O-horizon of the forest soils exerts a large influence. Hence the A-horizon represents--for p,p' DDT and gamma-HCH in particular--at pH values of 2.90 a pollutant reservoir which should not be underestimated and which could endanger the rhizosphere and the groundwater. According to PCBs, in the more acidic soils with a pH value <4.0 the lipophilic higher polychlorinated biphenyls were found to be more highly enriched in the humus layer. PMID- 11886090 TI - Effect of alcohol addition on the movement of petroleum hydrocarbon fuels in soil. AB - Groundwater contamination by fuel spills from aboveground and underground storage tanks has been of growing concern in recent years. This problem has been magnified by the addition of oxygenates, such as ethanol and methyl-tertiary butyl ether (MTBE) to fuels to reduce vehicular emissions to the atmosphere. These additives, although beneficial in reducing atmospheric pollution, may, however, increase groundwater contamination due to the co-solvency of petroleum hydrocarbons and by the provision of a preferential substrate for microbial utilisation. With the introduction of ethanol to diesel fuel imminent and the move away from MTBE use in many states of the USA, the environmental implications associated with ethanol additive fuels must be thoroughly investigated. Diesel fuel movement was followed in a 1-m soil column and the effect of ethanol addition to diesel fuel on this movement determined. The addition of 5% ethanol to diesel fuel was found to enhance the downward migration of the diesel fuel components, thus increasing the risk of groundwater contamination. A novel method using soil packed HPLC columns allowed the influence of ethanol on individual aromatic hydrocarbon movement to be studied. The levels of ethanol addition investigated were at the current additive level (approx. 25%) for ethanol additive fuels in Brazil and values above (50%) and below (10%) this level. An aqueous ethanol concentration above 10% was required for any movement to occur. At 25% aqueous ethanol, the majority of hydrocarbons were mobilised and the retention behaviour of the soil column lessened. At 50% aqueous ethanol, all the hydrocarbons were found to move unimpeded through the columns. The retention behaviour of the soil was found to change significantly when both organic matter content and silt/clay content was reduced. Unexpectedly, sandy soil with low organic matter and low silt/clay was found to have a retentive behaviour similar to sandy subsoil with moderate silt/clay, but little organic matter. It was concluded that sand grains might have a more important role in the adsorption of petroleum hydrocarbons than first realised. This method has shown that soil packed HPLC columns can be used to provide a quick estimate of petroleum hydrocarbon, and possibly other organic contaminant, movement in a variety of different soil types. PMID- 11886091 TI - Composition of human excreta--a case study from Southern Thailand. AB - In Thailand, human excreta might be recycled into agricultural soils as a supplement to commercial fertiliser and thereby enrich the general fertility of the soils. However, for Thailand an adequate knowledge of the quality of human excreta, in order to assess its fertiliser potential, is not available. A literature survey revealed only very limited information of the chemical composition and generation rate of human excreta in South East Asia. Data from other parts of the world also lacked specific information on collection and analytical methods, or the studies were typically 20-30 years old. In the present study the composition of human excreta has been studied in three case study areas in Southern Thailand: Kuan Lang, Phattalung and Prik. The inhabitants of the three areas represent people of Southern Thailand by age, sex, occupation, religion and type of residence. Human excreta was collected and stored for 1 week from five persons in each area, who each had their own toilet and collection bucket. In parallel, a septic tank at the Observation and Protection (O&P) Centre of Songkhla (a boys prison institution) adjacent to the three study areas was used as a daily sampling point, to obtain data on average amounts of human excreta and chemical composition. Information on average values of generation rate and chemical composition was obtained as well as inter-human variation. However, no significant variation was found between the results for human excreta at the O&P Centre or from the 15 individuals. Furthermore, there was no significant influence of age, sex, occupation or religion on the chemical composition. The only significant variation was that the older people excreted larger amounts of total wet matter than the younger, which could be due to a higher water intake, in order to reduce the risk of constipation. The generation rate found was 0.6-1.2 1 urine/cap/day and 120-400 g wet faeces/cap/day. The generation rate of the elements in the excreta was 7.6-7.9 g N/cap/day, 1.6-1.7 g P/cap/day, 1.8-2.7 g K/cap/day, 1.0-1.1 g S/cap/day, 0.75-1.5 g Ca/cap/day, 0.25 0.4 g Mg/cap/day, 9-16 mg Zn/cap/day, 1.4-1.5 mg Cu/cap/day, 0.3 mg Ni/cap/day, 0.02-0.03 mg Cd/cap/day, 0.07-0.14 mg Pb/cap/day, 0.01 mg Hg/cap/day and 0.8-1.1 mg B/cap/day. The metals (Ca, Mg, Zn, Cu, Ni, Cd, Pb, Hg) are mainly excreted via the faeces and the remaining elements (N, P, K, S, B) are mainly excreted via the urine. It can be concluded that human excreta constitutes a large fertiliser resource, which presently is not utilised in Thailand. PMID- 11886092 TI - Residual pollution load of soils impacted by the Aznalcollar (Spain) mining spill after clean-up operations. AB - By comparing total concentrations of potentially toxic elements in soils affected by the Aznalcollar mining spill with those of the adjacent unaffected soils, it can be inferred that after the sludge removal, there still exists a considerable amount of residual pollution. This exceeds the suitable levels for cultivation, especially in the case of arsenic for which total concentrations are in the range of values above which eco-toxicity is considered to be possible. Elemental distribution in the soil seems to be determined by two distinctive associations (As-Pb-Hg-Sb and Cu-Zn-Cd) with different geochemical behaviours. PMID- 11886093 TI - Evaluation of lead exposure in workers at secondary lead smelters in South Korea: with focus on activity of erythrocyte pyrimidine 5'-nucleotidase (P5N). AB - To evaluate lead exposure among secondary lead-smelting workers with a focus on erythrocyte pyrimidine 5'-nucleotidase (P5N) activity, blood lead concentration (PbB), activity of P5N and other biological variables were examined in 88 exposed workers in five secondary lead smelters and in 24 non-exposed workers in Korea. All of the mean values of air lead concentration (PbA) in the three processes, scrap pretreatment, blast furnace smelting, and refining and casting of the secondary lead smelters, markedly exceeded 0.05 mg/m3. In this survey, 29 (97%) of 30 air samples for lead exceeded 0.05 mg/m3. The highest mean PbA and PbB values were found in the section of blast furnace smelting. All of the mean PbB values in all the sections were higher than 30 microg/dl. PbB of 71 (81%) of the 88 exposed workers exceeded 30 microg/dl. In 31 (35%) of the exposed workers, PbB was above 60 microg/dl. Compared with the non-exposed group, zinc protoporphyrin (ZPP) in the exposed group was significantly increased, whereas erythrocyte P5N activity and activity of erythrocyte delta-aminolevulinic acid dehydratase (ALAD) were significantly inhibited. Erythrocyte P5N activity had valid biological correlation with PbB and with other biological variables, such as ALAD activity or ZPP. Lead exposure affected hemoglobin levels via inhibition of P5N activity, as well as the heme biosynthetic pathway, in the high-exposure state. PMID- 11886095 TI - Bryophytes as indicators of trace metals pollution in the River Brenta (NE Italy). AB - The performance of aquatic bryophytes for detecting metal pollution was tested along the River Brenta, in NE Italy. Nine sampling sites were selected, three of them along short tributaries with no anthropic influence upstream, the others along the main bed of the river. Two sites were deliberately placed downstream from a previously known pollution focus. The multivariate analysis (classification and ordination) of the matrix of 10 metals and 38 samples revealed: (i) a good discrimination between 'clean' and potentially polluted sites; (ii) two contamination syndromes, one by As and, to a lesser degree, Pb, and the other by Cr, due to agricultural and industrial activities, respectively; and (iii) the previously known pollution focus was clearly detected. The magnitude of contamination was estimated by means of a comparison between local backgrounds and concentrations of metals measured. The distance of aquatic bryophytes from the center of the river was negatively correlated with metal concentrations, which suggests that this factor should be taken into account in the implementation of sampling protocols. PMID- 11886094 TI - Modelling long-term stream acidification in the chemically heterogeneous Upper Severn catchment, Mid-Wales. AB - A two-box version of the long-term acidification model MAGIC is applied to the Upper Severn catchment, Mid-Wales. Comparison between modelled output and the observed stream- and groundwater chemistry points to the limitations of modelling, due to the inherent complexity and variability in the catchment hydrology, soils, geology and chemistry. The MAGIC model is used to produce long term hind- and forecast predictions of average stream-, soil- and groundwater chemistry, and to simulate long-term changes in sulfate and nitrate deposition, in line with current proposals of reduction. Changes in flow-routing pathways between soil- and groundwater are simulated and the long-term effects on streamwater quality noted. The use of a long-term acidification model enabled the simulation of streamwater quality under these different case scenarios. However, the modelled output is insensitive to depositional and flow routing changes, indicating that catchment processes are not being represented to a sufficient degree. Changes in simulated output as a result of increased acidic deposition are not statistically significant, lying within the variance of long-term observed data. Simulated changes in flow routing suggest a lack of model sensitivity, in terms of the effect on stream chemistry. The need for large amounts of measured data to ensure correct model representation of the hydrology, chemistry and the heterogeneous/variable nature of upland catchments is outlined. It is vital that these long-term data are available to ensure that problems do not arise due to over-reliance by catchment managers on potentially unreliable modelled output. PMID- 11886096 TI - Scanning laser ablation-ICP-MS tracking of platinum group elements in urban particles. AB - While it has now been demonstrated that platinum group elements (PGE) are released from automobile catalysts into the environment, less is known about the form in which they are emitted and transported. Here we show that scanning laser ablation-inductively coupled plasma-mass spectrometry (scanning laser ablation ICP-MS) can identify and track individual particles released from automobile catalysts present in environmental particulates and sediments. Particles with high PGE concentrations were found in the exhaust of gasoline and diesel vehicles equipped with catalytic converters. The PGE-Ce association in individual particles provides a definitive fingerprinting for tracking catalyst particles in environmental compartments, while relative PGE signal intensity is an indication of the catalyst type. Scanning laser ablation-ICP-MS of road and aquatic sediments revealed a few PGE containing catalyst particles and it was possible to identify catalyst types for the origin of these particles. PMID- 11886097 TI - Lichen (Xanthoria parietina) biomonitoring of trace element contamination and air quality assessment in Pisa Province (Tuscany, Italy). AB - In the northern part of Pisa Province (Tuscany, Italy), the use of lichens as both airborne trace element biomonitors and air quality bioindicators is described. The following elements were analysed in Xanthoria parietina: As, Cd, Cr, Ni, Pb, V, Zn and Hg, and the results are compared with those we previously obtained in Livorno Province (Tuscany) using the same lichen species. The results identify spots of different environmental metal contamination and air quality. Median values of Pb, V and Ni concentrations were much lower than those of Livorno Province, with maximum values even nine times lower. Arsenic contamination was also lower, while Cd, Hg and Zn levels were similar in the two areas. In Pisa Province, the highest levels of contamination were recorded for Zn, Pb, Cr and Ni, and a degree of agreement was found between air quality and metal concentrations in lichens. The air quality in Pisa Province is better than in Livorno Province, even if the different climatic and orographic features of the two areas may influence the presence of lichen species and thus an assessment of air quality. PMID- 11886098 TI - Air quality and well-being perception in subjects attending university libraries in Modena (Italy). AB - We studied four libraries in the University of Modena and Reggio Emilia (Northern Italy) to determine the presence of polluting agents such as total dusts, formaldehyde and other volatile organic compounds (VOCs) including benzene, toluene and xylenes and to assess the sense of well-being perceived by library users. This investigation was suggested by an increase in reported symptoms related to Sick Building Syndrome (SBS) observed in recent decades among people spending most of their time in various indoor environments, including libraries. The microclimatic conditions and the concentrations of pollutants indicated an acceptable situation on the whole, even though a wide range of total dust values (40-350 microg/m3) and total VOCs (203-749 microg/m3) was observed. However, the perception of the different environmental parameters by the 130 library users that were interviewed identified the existence of some discomfort mainly caused by the feeling of poor ventilation. Moreover, 78.5% of the subjects stated they had at least one of the 16 investigated symptoms potentially related to a SBS. The place of occurrence of the self-reported symptoms was also investigated, the symptoms arising during library attendance more frequently than elsewhere were only four, and in particular feeling hot, sore eyes, dry throat and breathing difficulties. Overall, our study has shown the existence of an association between microclimatic perceptions as reported by library users and subjective symptoms related to SBS, considering the four libraries separately, the highest prevalence of self-reported symptoms was found in the library where environmental discomfort as perceived by users was greatest. Nevertheless, an association between subjective self-reported symptoms and both microclimatic conditions as resulted by instrumental measurements and/or pollutants concentrations was not apparent. PMID- 11886099 TI - Assessment of bioavailable arsenic and copper in soils and sediments from the Antofagasta region of northern Chile. AB - Copper levels of nearly 500 mg l(-1) were measured in aqueous extracts of soil and sediment samples from the lowlands of Antofagasta. Arsenic levels of up to 183 mg l(-1) were found in river sediments, and 27.5 mg l(-1) arsenic was found at the location of a dam where potable water is extracted. This indicates that the arsenic contamination of water supplies reported recently for the pre-Andes may be a widespread problem throughout the region. Copper contamination from smelting activities also provides cause for concern as elevated levels were found in aqueous extracts of soil up to 20 km away from a smelter. This study went beyond traditional chemical analysis by assessing the potential benefits of using microbial biosensors as an alternative to determination of chemical speciation, to provide an environmentally relevant interpretation of soil/sediment residue levels. This approach is simple to use and enables a rapid, low cost assessment of pollutant bioavailability. It may, therefore, be of use for further investigations in the region and beyond. PMID- 11886101 TI - Dry deposition profile of small particles within a model spruce canopy. AB - Data on dry deposition of 0.82 microm MMAD uranium particles to a small scale, 'model' Norway spruce (Picea abies) canopy have been determined by means of wind tunnel experiments. These are presented for both the total canopy and for five horizontal layers within the canopy. The results show a complex pattern of deposition within the canopy. The highest deposition velocity Vg (0.19 cm s(-1)) was recorded for the topmost layer within the canopy (i.e. the layer in direct contact with the boundary layer) whereas the lowest Vg (0.02 cm s(-1)) occurred at the soil surface. Vertical penetration of depositing aerosol through the canopy was influenced by variations in biomass, wind velocity and turbulence within the canopy. A total canopy Vg of 0.5 cm s(-1) was obtained and this is in line with field measurements of Vg reported in literature for both anthropogenic and radionuclide aerosols of similar size ranges. Extrapolation of wind tunnel data to 'real' forest canopies is discussed. The information presented here is of importance in predicting the likely contribution of dry deposition of aerosols to pollutant inputs to forest ecosystems, particularly in the context of radioactive aerosol releases from nuclear installations. The application of the present data may also be appropriate for other pollutant aerosols such as SO4, NO3 and NH4, which are characterised by particle sizes in the range used in this study. PMID- 11886102 TI - Biogeochemical dynamics following land use change from forest to pasture in a humid tropical area (Rondjnia, Brazil): a multi-element approach by means of XRF spectroscopy. AB - Forest burning for pastures in tropical areas represents an important component of biogeochemical cycles. In order to provide information concerning chemical modifications after forest burning, in this local study the total contents of 29 elements in topsoils were analyzed when forest is changed to pasture land. The work was carried out in 1999 in Rondjnia state (Brazilian Amazon Basin) focussing on a native forest site and four neighboring pastures established in 1987, 1983, 1972 and 1911 after forest conversion. Chemical fingerprint graphs of the pasture soils related to the forest soil illustrated mainly higher contents for the vast majority of macro- and micro nutrients, but for other elements as well (e.g. Ba, Sr, Cr, Ni, V or Pb). Also increases of pH levels were measured in all pastures, which remained higher than the forest values for decades. After initial increases of most of the elements in pasture of 1987 the decreases of some macro elements (e.g. C, N, K, Mg, S) in pasture 1983 as well as again the enhanced levels in pasture 1972 and 1911 suggest both a persistent leaching of these elements and a function of pasture age where external element inputs exceed outputs. Ash deposition, accumulation of organic matter, animal excreta as well as natural soil conditions are discussed as influencing factors on the element contents of the original forest and the pasture soils. Nevertheless, in this particular area continuous pasturing after forest clearing primarily enriched the soils in elements. PMID- 11886100 TI - Alterations of heme metabolism in lymphocytes and metal content in blood plasma as markers of diesel fuels effects on human organism. AB - Workers in the diesel fuel distribution trade are intensively exposed to fuel vapours. Diesel fuel presents the main source of air pollution by benzene at a marine diesel fuel terminal. Levels of benzene are used to evaluate the external exposure to diesel fuel. Since benzene causes alterations in porphyrin metabolism, and some of these may lead to the generation of tumours, heme synthesis is proposed as a biomarker of early health effects of diesel fuel. A group of 20 workers exposed to diesel fuel and a group of 20 unexposed persons were examined and interviewed using structured questionnaires. The levels of 5 aminolevulinic acid (ALA) and protoporphyrin (PP), activities of ALA synthase and ferrochelatase, as well as levels of PP associated with DNA were determined in lymphocytes spectrophotometrically. Amounts of the metals Cd, Mn, Zn, Cu and Ca were measured in blood plasma by flame atomic absorption spectrometry method. Both ALA and PP levels were significantly increased in marine terminal workers: 3.0 +/- 0.4 vs. 0.8 +/- 0.2 nmol/10(6) lymphocytes: and 511 +/- 164 vs. 389 +/- 77 pmol/10(6) lymphocytes in exposed and control individuals, respectively. ALA synthase activity was 2.5 fold higher in lymphocytes of workers exposed to diesel fuels (P < 0.01). At the same time ferrochelatase activity was decreased and protoporphyrin level was accordingly elevated. The amount of porphyrin associated with DNA increased 1.4 fold in exposed workers (P = 0.05). Among all investigated metals in blood plasma of exposed workers only zinc levels were statistically significantly increased (P < 0.05). The disturbances of heme metabolism in lymphocytes and zinc level in blood plasma caused by diesel fuel exposure seems to be a useful biomarkers for carcinogenic risk assessment. PMID- 11886103 TI - Tracheal dyskinesia associated with midline abnormality: embryological hypotheses and therapeutic implications. AB - Abnormalities of tracheal rigidity, which may lead to the collapse of the airway during expiration and consequent complications, characterize two groups of disorders: tracheomalacia (weakness of the anterior cartilaginous arc of the trachea) and tracheal dyskinesia (dysfunction of the posterior membranous trachea). Tracheal dyskinesia can either be isolated or associated with a more complex syndrome of malformations: esophageal atresia, tracheoesophageal fistula and laryngotracheal cleft. Although our knowledge of the embryological development of the tracheoesophageal axis remains limited, the existence of these associations suggests that tracheal dyskinesia is of congenital origin. The presentation of three clinical cases demonstrates that the coexistence of a midline malformation and of tracheal dyskinesia complicates the therapeutic management of the first malformation. In particular, the postoperative follow-up is often more difficult, and a long-term tracheostomy is often required (sometimes for several years). However, it must be pointed out that tracheal dyskinesia, even in the associated forms, has a good long-term prognosis, since spontaneous resolution as the child grows up is the rule. PMID- 11886104 TI - Management of respiratory distress syndrome: an update. PMID- 11886105 TI - Persistent pulmonary hypertension in preterm infants with respiratory distress syndrome. PMID- 11886106 TI - Bronchopulmonary dysplasia: prenatal and postnatal influences. PMID- 11886107 TI - Central chemoreceptor function in children. PMID- 11886108 TI - Abnormalities of the chemical control of breathing: Clinical correlates in infants and children. AB - Abnormalities of the chemical control of breathing may go unrecognized and lead to life-threatening events, especially during sleep. Tests to assess chemical control in vivo have not yet been standardized, and their results may be difficult to interpret. Non-invasive monitoring of gas exchange and polysomnography are essential to assess the severity of hypoventilation and the extent to which it is dependent on the state of alertness. One has to be aware that some patients may have increased vulnerability to stress, and that mild infections may trigger acute hypoventilation. To date, no pharmacological approaches have proved effective in the long-term. Therefore, the management of infants and children with abnormal chemical control of breathing includes ventilatory support during sleep and diaphragmatic pacing during wakefulness, if necessary. Further research is needed to improve our understanding of the mechanisms controlling chemosensitivity and of the developmental plasticity of chemosensitivity during infancy and childhood. Genetic influences, as well as environmental factors in utero or during early infancy, may contribute to abnormal chemical control of breathing during infancy and childhood. PMID- 11886109 TI - Imaging of congenital lower respiratory tract malformations: prenatal diagnosis by magnetic resonance imaging. PMID- 11886110 TI - Prenatal ultrasound diagnosis of congenital lung lesions. PMID- 11886111 TI - Postnatal imaging of congenital lower respiratory tract malformations. PMID- 11886112 TI - Management of childhood asthma in Indonesia. PMID- 11886113 TI - Specific problems with the diagnosis and therapy of asthma in Colombia. PMID- 11886114 TI - The special challenges of aerosolization of medication to technologically dependent children. PMID- 11886115 TI - What are the problems with the NIH guidelines? What are the solutions? PMID- 11886116 TI - Home therapy: oxygen and nutrition. PMID- 11886117 TI - Tracheostomy in chronic lung disease: care and follow-up. PMID- 11886118 TI - Lung function testing: infants on ventilatory support. PMID- 11886119 TI - Lung function testing: chronic lung disease of infancy. PMID- 11886120 TI - Asthma and lung function. PMID- 11886121 TI - Mechanisms involved in cystic fibrosis airway inflammation. PMID- 11886122 TI - The effects of postnatal environment. PMID- 11886123 TI - Has the role of atopy in the development of asthma been over-emphasized? PMID- 11886124 TI - Lower respiratory tract infection in Sub-Saharan Africa: epidemiology, diagnostic, and therapeutic strategy. PMID- 11886125 TI - Respiratory infections in Morocco: past, present, and future. PMID- 11886126 TI - Diagnostic and therapeutic problems of lower respiratory tract infections in Turkey. PMID- 11886127 TI - Lower respiratory tract infections in developing countries. PMID- 11886128 TI - Difficulties in the diagnosis and treatment of acute lower respiratory tract infections in Uruguay. PMID- 11886129 TI - Asthma education: the impact on care: the Quebec experience. PMID- 11886130 TI - Changing pattern of lower respiratory tract infections in Hong Kong children. PMID- 11886131 TI - Pressurized metered dose inhalers: optimal use of hydrofluoroalkane devices. PMID- 11886132 TI - Biology of respiratory epithelial cells: role in defense against infections. PMID- 11886133 TI - Impact of specialty care on cost containment and pulmonary function. PMID- 11886134 TI - Nutrition and severe chronic respiratory diseases: pathophysiologic mechanisms. PMID- 11886135 TI - Nutrition and severe chronic respiratory disease. PMID- 11886136 TI - Nutrition and severe chronic respiratory diseases: the pediatric experience. PMID- 11886138 TI - Epidemiology of asthma in Maghrebian countries. PMID- 11886137 TI - The epidemiology of asthma and allergic diseases: a comparison between Eastern and Western European countries. PMID- 11886139 TI - Follow-up of asthma from childhood to adulthood. PMID- 11886140 TI - Epidemiology of asthma in Portugal, Cape Verde, and Macao. PMID- 11886141 TI - Epidemiology of food allergy in Portugal. PMID- 11886142 TI - Allergy to lentils in Spain. PMID- 11886143 TI - Mustard allergy in children. PMID- 11886144 TI - Pollen allergy in Israel. PMID- 11886145 TI - Exercise and food-induced anaphylaxis. PMID- 11886146 TI - Bronchopulmonary dysplasia: thirty-three years later. PMID- 11886147 TI - Epidemiology of cigarette smoking in adolescents: the German experience. PMID- 11886148 TI - Position-centered national coalition: how to build a coalition in connection with a public health campaign to obtain tobacco control measures. PMID- 11886149 TI - Theoretically sound approaches to prevention and cessation of cigarette smoking among youths. PMID- 11886150 TI - Worldwide epidemiology of tuberculosis. PMID- 11886151 TI - Usefulness of serological tests in childhood TB. PMID- 11886152 TI - Treatment of pediatric tuberculosis in the year 2000. PMID- 11886154 TI - The spectrum of interstitial lung disease in childhood. PMID- 11886153 TI - New strategies of tuberculosis control in the developing world. PMID- 11886155 TI - Persistent tachypnea of infancy (PTI)--a new entity. PMID- 11886156 TI - Diffuse interstitial lung disease in infants. PMID- 11886157 TI - Regulation of alveolar growth. PMID- 11886158 TI - Tracheomalacia associated with compressive cardiovascular anomalies in children. PMID- 11886159 TI - Alveolar growth: development disorders. PMID- 11886160 TI - Prevention of pneumococcal respiratory infections. PMID- 11886161 TI - Prevention in respiratory syncytial virus infections. PMID- 11886162 TI - Histopathology of fatal asthma: drowning in mucus. PMID- 11886163 TI - Therapeutic strategies in near fatal asthma in children. AB - Status asthmaticus in children is associated with non-decreasing morbidity and mortality. Oxygen, beta2-agonists and corticosteroids remain the mainstay of therapy. Other therapies may be of help although their efficacy remains anecdotal. The development of new methods of ventilation for children with status asthmaticus is of utmost importance. PMID- 11886165 TI - Perspectives in gene therapy. PMID- 11886164 TI - Novel approaches to inflammation and infection in the cystic fibrosis lung. PMID- 11886166 TI - High level expression and purification of the Epstein-Barr virus encoded cytokine viral interleukin 10: efficient removal of endotoxin. AB - To characterize the structural and functional properties of viral interleukin 10 (vIL-10), its cDNA was cloned into the bacterial expression vector pMAL-c2, which directs the synthesis of the inserted gene as a fusion protein with maltose binding protein (MBP). The MBP-vIL-10 fusion protein was expressed in Escherichia coli and purified from cell lysates using amylose resin chromatography. Viral interleukin 10 (IL-10) was released from the fusion protein by cleavage with the proteolytic enzyme factor Xa. We show that vIL-10 will bind to heparin and use this property to purify vIL-10 from factor Xa cleaved products and trace contaminants using heparin agarose chromatography. A simple one-step procedure is described for the removal of endotoxins from heavily contaminated vIL-10 preparations. The protocol exploits the high binding affinity of MBP for amylose resin or vIL-10 for heparin and the ability of Triton-X114 to dissociate endotoxins from proteins. The biological activity of purified vIL-10 was demonstrated through its ability to inhibit interferon gamma (IFN-gamma) production by mitogen activated peripheral blood mononuclear cells and to down regulate HLA-class II expression on activated monocytes/macrophages. The availability of an efficient expression and purification strategy for vIL-10 together with appropriate assays will contribute to a greater understanding of how vIL-10 has evolved to retain and modify those activities of cellular IL-10 best suited for Epstein-Barr virus (EBV)'s specialized niche within the host. PMID- 11886167 TI - Stabilization of hypoxia-inducible factor-1alpha is involved in the hypoxic stimuli-induced expression of vascular endothelial growth factor in osteoblastic cells. AB - It has been suggested that blood vessel formation is an important event coupled to bone formation. The expression of vascular endothelial growth factor (VEGF), a potent angiogenic factor, has been shown to be greatly stimulated in osteoblasts by hypoxic stimuli such as deprivation of oxygen and treatment with cobalt. In other cell types, hypoxia-inducible factor-1 (HIF-1) that binds hypoxia-response element (HRE) has been shown to mediate gene expression induced by hypoxic stimuli. In this study, we investigated the effects of hypoxic stimuli on HIF-1, HRE, and VEGF in osteoblastic cell lines. Exposure of these cells to hypoxia or cobalt resulted in a great increase in the protein level of HIF-1alpha and the gene expression of VEGF. Transforming growth factor-beta1, prostaglandin E2, dexamethasone, and 1,25-dihydroxyvitamin D3 that have been shown to regulate VEGF gene expression in osteoblasts had no effect on HIF-1alpha induction. Blocking the enzymatic activity of phosphatidylinositol 3-kinase, p38, MEK-1 did not have any effect on the cobalt-stimulated increase of HIF-1alpha in these cells. In contrast, N-acetylcysteine (NAC), a scavenger of reactive oxygen species, abolished the cobalt induction of HIF-1alpha and that of the VEGF and a HRE driven reporter genes. However, the hypoxia responses were not affected by NAC. These findings suggest that hypoxia and cobalt can induce VEGF gene expression in osteoblasts by increasing the level of HIF-1alpha protein through different mechanisms. PMID- 11886168 TI - Interleukin (IL)-4 inhibits phorbol-ester induced HIV-1 expression in chronically infected U1 cells independently from the autocrine effect of endogenous tumour necrosis factor-alpha, IL-1beta, and IL-1 receptor antagonist. AB - The anti-inflammatory cytokine interleukin 4 (IL-4) has shown both inductive and inhibitory effects on the replication of the human immunodeficiency virus type 1 (HIV-1) in primary CD4+ T cells and mononuclear phagocytes. In this study, IL-4 did not induce virus production, but inhibited phorbol esters (PMA)-stimulated HIV expression in chronically infected promonocytic U1 cells. This effect, however, was not accounted for by a decreased secretion of endogenous TNF-alpha induced by phorbol myristate acetate (PMA). We also observed that PMA upregulated the production of both IL-1beta and of IL-1 receptor antagonist (IL-1ra). IL-4 inhibited the secretion of IL-1beta and strongly increased that of IL-1ra; however, these effects were not responsible of IL-4-mediated inhibition of PMA induced HIV expression since anti-IL-1ra antibodies did not revert IL-4 mediated suppression. U1 cells were transiently transfected with both wild-type (WT) long terminal repeat (LTR) constructs, or with LTR plasmids containing deletions of either the NF-kappaB or the Sp-1 binding sites. IL-4 inhibited LTR-driven transcription triggered by PMA stimulation of U1 cells, and this effect was dependent upon intact NF-kappaB but not Sp-1 binding sites. Thus, IL-4 may favour a state of microbiological quiescence in infected monocytic cells bypassing the induction of HIV expression mediated by pro-inflammatory cytokines. PMID- 11886170 TI - Combination interleukin-2 and interleukin-12 induces severe gastrointestinal toxicity and epithelial cell apoptosis in mice. AB - Interleukin 2 (IL-2) and interleukin 12 (IL-12) have potent anti-tumour activity as single agent therapy against several different murine and human tumours. Combining these cytokines may result in improved therapeutic effectiveness, however, the toxicity associated with simultaneous administration is prohibitive. This study was designed to determine the specific histopathologic changes associated with combination therapy. Mice were treated with 5 days of interleukin 2, interleukin-12, or both using standard doses and schedules. Histologic specimens were prepared from all internal organs on a daily basis to identify specific pathologic abnormalities. Treatment with interleukin-2, interleukin-12, or both resulted in pathologic insult to the liver and gastrointestinal tract. Mild lymphoplasmacytic infiltrates were seen in the liver. The most significant pathology was seen in the large bowel and consisted of apoptosis of colonic epithelial cells. While recovery of injured gastrointestinal mucosa occurred in mice treated with interleukin-2 or interleukin-12 alone, combination therapy resulted in death before recovery was possible. Combination interleukin-2 and interleukin-12 therapy results in irreversible injury of the colon as manifested by increased epithelial cell apoptosis and death in mice. Understanding the pathologic changes associated with combination cytokine therapy may lead to strategies that prevent toxicity while maintaining therapeutic effects. PMID- 11886169 TI - Bone morphogenetic proteins act synergistically with haematopoietic cytokines in the differentiation of haematopoietic progenitors. AB - We examined the effects of bone morphogenetic protein-2 (BMP-2), -3, -4, -5, -6, and -7 on the proliferation and differentiation of bone marrow CD34+ haematopoietic progenitors in semi-solid medium. The BMPs had no effect on haematopoietic colony development when added to medium containing erythropoietin (Epo) or Interleukin-3 plus Epo. Synergistic effects with the haematopoietic cytokines stem cell factor (SCF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were observed. In conjunction with GM-CSF and Epo, BMP-4 increased the number of both erythroid and granulocyte/monocyte colonies formed in semi-solid medium (P<0.01). No other BMP stimulated erythroid colony development under these conditions, while BMP-3, BMP-7 (P<0.01), BMP-5, and BMP-6 (P<0.05) stimulated granulocyte/monocyte colony formation. BMP-7 acted synergistically with stem cell factor to increase granulocyte/monocyte colony formation but not erythroid colony formation. The other BMPs did not affect either erythroid or granulocyte/monocyte colony development under these conditions. These results suggest that individual BMPs form part of the complement of cytokines regulating the development of haematopoietic progenitors, and in particular, point to a role for BMP-4 in the control of definitive, as well as embryonic erythropoiesis. PMID- 11886171 TI - Profiles of pro-inflammatory cytokines in the serum of rabbits after experimentally induced acute pancreatitis. AB - In a recent study we have demonstrated that interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF-alpha) serum levels correlate positively with the severity of acute pancreatitis (AP), induced by bile acid injected into the pancreatic duct of rabbits. In this article we describe the effect of an IL-10 analogue IT9302 and a monoclonal anti-IL-8 (mon. IL-8) antibody on the content of several pro-inflammatory cytokines in the serum of rabbits, after induction of AP. We found that the serum content of inflammatory cytokines IL-8, IL-1beta, TNF alpha and monocyte chemoattractant protein 1 (MCP-1) are increased during AP. Injection of IT9302 or mon. IL-8 antibody, diminish the concentration of these cytokines in the serum, with the exception that mon. IL-8 antibody actually increased the circulating level of MCP-1. In addition, intravenous administration of IT9302 increased the serum levels of IRAP, an IL-1beta receptor antagonistic cytokine. Furthermore, intravenous injection of mon. IL-8 antibody increased serum levels of IL-4. It can be concluded that both the human IL-10 analogue IT9302 and mon. IL-8 antibody are able to alter the pro-inflammatory cytokine levels in rabbits suffering from experimentally induced AP. PMID- 11886172 TI - Plasma and urinary cytokine homeostasis and renal function during cardiac surgery without cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) significantly contributes to the plasma pro inflammatory cytokine response at cardiac surgery. Complementary plasma and urinary anti-inflammatory cytokine responses have been described. The pro inflammatory cytokines interleukin 8 (IL-8), tumour necrosis factor alpha (TNF alpha) and interleukin 1beta (IL-1beta) have lower molecular weights than the anti-inflammatory cytokines interleukin 10 (IL-10), interleukin 1 receptor antagonist (IL-1ra) and TNF soluble receptor 2 (TNFsr2) and thus undergo glomerular filtration more readily. In vitro work suggests that proximal tubular cells are vulnerable to pro-inflammatory cytokine mediated injury. Accordingly, this study investigated the hypothesis that cardiac surgery without CPB would not have significant changes in plasma and urinary cytokines and proximal renal dysfunction. Eight patients undergoing coronary artery bypass grafting (CABG) without CPB were studied. Blood and urine samples were analysed for pro- and anti inflammatory cytokines. Proximal tubular dysfunction was measured using urinary Nu-acetyl-beta-D-glucosaminidase (NAG)/creatinine and alpha(1) microglobulin/creatinine ratios. Plasma IL-8, IL-10, IL-1ra and TNFsr2 were significantly elevated compared with baseline. Urinary IL-1ra and TNFsr2 were significantly elevated, as were urinary NAG/creatinine and alpha(1) microglobulin/creatinine ratios. Two hours following revascularization, urinary IL-1ra correlated with urinary alpha(1)-microglobulin/creatinine ratios (P<0.05). As previously reported in CABG surgery with CPB, we now report that non-CPB cardiac surgery also has significant changes in plasma and urinary cytokine homeostasis and early proximal tubular injury. The correlation between urinary IL 1ra and alpha(1)-microglobulin/creatinine ratios is consistent with earlier suggestions of a mechanistic link between cytokine changes and proximal tubular dysfunction. The relative roles of CPB and non-CPB processes in producing inflammation still require definition. PMID- 11886173 TI - Molecular cloning of an IL-8-like CXC chemokine and tissue factor in rainbow trout (Oncorhynchus mykiss) by use of suppression subtractive hybridization. AB - Suppression subtractive hybridization (SSH) was performed to construct a Rainbow trout cDNA library enriched in sequences up-regulated in head kidney leukocytes after lipopolysaccharide (LPS) and tumour necrosis factor alpha (TNFalpha) stimulation. Random sequencing of fifty clones allowed the identification of a Rainbow trout interleukin 8 (IL-8)-related CXC chemokine, as well as the Rainbow trout tissue factor (TF) precursor. Expression of both the IL-8-like chemokine and TF is induced after LPS and TNFalpha stimulation, indicating that they are associated with inflammatory responses in fish, as has been suggested in mammals. These results confirm the potential of SSH to identify cytokines and immuno regulatory genes in fish. PMID- 11886174 TI - Cloning of a novel rainbow trout (Oncorhynchus mykiss) CC chemokine with a fractalkine-like stalk and a TNF decoy receptor using cDNA fragments containing AU-rich elements. AB - An activation-specific cDNA library was made from phytohaemagglutinin (PHA) activated haematopoietic cells of the rainbow trout (Oncorhynchus mykiss) using the technique of suppression subtractive hybridization. Several immune system genes were identified, including an interleukin (IL)1 receptor related protein and two invariant chain-like proteins. Many clones showed no similarity by BLAST search, but had AU-rich elements. These fragments were labelled and used for hybridization with a PHA-activated head kidney cDNA library. Several immune system genes were isolated by this technique, including a tumour necrosis factor (TNF) decoy receptor and a novel chemokine, designated trout chemokine 2. The TNF receptor is 285 amino acids in length and is 32-36% identical to a brook trout and human homologue. The CC chemokine is 44% identical at the amino acid level to a carp CC chemokine and approximately 20% identical to several mammalian CC chemokines. However, it has a 91 amino acid stalk-like structure at its COOH end, which is similar to the glycosylated stalk of fractalkine, a mammalian CX(3)C chemokine. In summary, AU-rich fragments obtained from an activation-specific library proved useful as hybridization probes for isolating trout immune system genes. PMID- 11886175 TI - Oligomerization of IL-2Ralpha. AB - Interleukin (IL) 2 receptor subunit alpha (IL-2Ralpha) increases the affinity of the IL-2 receptor complex while hetero-association of IL-2Rbeta and gamma(c) chains initiates a proliferative signal. We show here that IL-2Ralpha is necessary for receptor clustering required for augmentation of IL-2 signalling. Cells expressing chimeras incorporating the extracellular domain of IL-2Ralpha demonstrated IL-2 independent homo-association of the IL-2Ralpha chimera. Singly or co-transfected IL-2Rbeta and gamma(c) chimeras showed no spontaneous or IL-2 inducible oligomerization. Co-transfection of IL-2Ralpha and IL-2Rbeta (+/- gamma(c)) chimeras diminished spontaneous IL-2Ralpha chimera oligomerization and permitted IL-2-inducible hetero-oligomerization of receptor components. Homo association of IL-2Ralpha was also demonstrated by fluorescence resonance energy transfer (FRET). The spontaneous homo-oligomerization property of IL-2Ralpha required the membrane proximal region of the receptor (exon 6) by deletion analysis; the IL-2 inducible oligomerization property of IL-2Ralpha required the second "sushi" domain (exon 4). This work provides insight into the mechanics of this complex receptor system and to other receptor complexes in the immune system that send signals by clustering receptor subunits. PMID- 11886176 TI - Role of pRB-family/E2F complex in the inhibition of IL-3-dependent lymphoid cell proliferation. AB - Interleukin 3 (IL-3)-dependent proliferation of haematopoietic cells is specifically inhibited by p130, a member of the pRB-family proteins. p130 interacts with the cell-cycle regulatory E2F transcription factors, notably E2F-4 and E2F-5, and affects promoters containing E2F-binding sites through two distinct mechanisms. First, upon complex formation with E2F, it blocks transcriptional activation by E2F. Second, the formed p130-E2F complex binds to E2F sites and actively represses transcription by inhibiting the activity of surrounding enhancer elements on the promoter. To pursue the relative contributions of each mechanism in the p130-mediated inhibition of IL-3-dependent cell proliferation, we employed a dominant-negative DP-1, which suppresses both E2F-dependent transactivation and the formation of active transcriptional repressors. Ectopic expression of the dominant negative DP-1 in the IL-3 dependent BaF3 lymphoid cells gave rise to an inhibition of cell proliferation, which was concomitantly associated with a decrease in levels of cyclin E, an indispensable molecule for G1 to S-phase cell-cycle progression. Our results indicate that blocking E2F-dependent transactivation, but not the formation of p130-E2F transcriptional repressor complexes, is responsible for the inhibition of IL-3-dependent cell growth by p130. PMID- 11886177 TI - A possible role of IL-1ra 3'-untranslated region in modulation of protein production. AB - Human interleukin 1 (IL-1) receptor antagonist (hIL-1ra), an anti-inflammatory cytokine and naturally occurring antagonist of IL-1, may be regulated at both the transcriptional and post-transcriptional levels. The aim of this study was to evaluate post-transcriptional regulation of hIL-1ra with specific focus on the 3' untranslated region (3'-UTR) of hIL-1ra mRNA using the luciferase reporter gene system. Constructs were created containing the luciferase reporter gene followed by the hIL-1ra 3'-UTR or its modified variants. In monocyte/macrophage cell lines RAW264.7 and U937, the presence of the hIL-1ra 3'-UTR resulted in a 5.7-fold (n=6, P<0.001) and a 3.9-fold (n=7, P<0.001) decrease in transient reporter gene expression, respectively, with only a less than 2-fold mean difference in steady state mRNA levels in the former case. In a cell-free translation system, the presence of the middle segment of the 3'-UTR caused a 5.2-fold (n=5, P<0.001) decrease in the amount of luciferase synthesized. In contrast to synthetic 3'-UTR and that derived from bovine growth hormone RNA, the presence of hIL-1ra 3'-UTR resulted in accumulation of unprocessed transcripts in transfected cells. We conclude that hIL-1ra synthesis may be regulated at the post-transcriptional level through mechanisms involving the 3'-UTR of IL-1ra transcripts. PMID- 11886178 TI - Serum tumour necrosis factor-alpha and transforming growth factor-beta levels in chronic hepatitis C patients are immunomodulated by therapy. AB - Our aims were: (i) to characterize serum levels of tumour necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) in non-cirrhotics with hepatitis C; (ii) to correlate levels of theses cytokines with degree of disease at baseline; (iii) to characterize the immunomodulatory effects of therapy with response and (iv) to compare profiles of cytokines in patients treated with pegylated-interferon alpha-2b monotherapy (PMT) vs its combination with ribavirin (PCT1-low dose ribavirin and PCT2-high dose ribavirin). We studied 56 patients that were part of two randomized, controlled, clinical trials. At baseline, high TNF-alpha levels paralleled the degree of inflammation as determined by histology. In PCT2, a significant reduction was seen in levels of TNF-alpha, TGF beta and fibrosis scores when comparing baseline with follow-up. In sustained responders, regardless of therapy, the histological activity scores were lower at follow-up as compared to baseline. In conclusion, PCT2 is able to constantly reduce and sustain TNF-alpha levels, which is responsible for the sustained decline in liver inflammation as shown by the histological activity index and it is also able to reduce fibrosis as judged both by TGF-beta levels and fibrosis scores. PMID- 11886179 TI - The Berlin declaration on gold mining. Further observations and comments on the cyanide process to produce gold. PMID- 11886180 TI - Acute toxicity of lead on tolerance, oxygen consumption, ammonia-N excretion, and metal accumulation in Penaeus indicus postlarvae. AB - The estuaries and backwaters that are the potential breeding grounds of penaeid shrimps are subject to heavy metal pollution through industrial effluents and domestic sewage. In the present investigation, laboratory experiments were conducted to study the acute toxicity of lead on tolerance, oxygen consumption, ammonia-N excretion, and metal accumulation in Penaeus indicus postlarvae. Static bioassay tests were employed to determine tolerance limits. Oxygen consumption, ammonia-N excretion, and metal accumulation were determined in postlarvae by exposing them to different concentrations of lead for a period of 48 h. Oxygen consumption measurements were made by using a respiratory chamber equipped with an oxygen electrode and ammonia-N was determined with trione (dichloro-S-triamine 2,4,6(1H,3H,5H-trione)). Accumulation of metal was estimated by wet-ash method. The LC50 value for 96 h was 7.223 ppm and the regression equation Y=4.1638+0.9738X with correlation coefficient of 0.9613 was obtained by probit method. A decrease in oxygen consumption and ammonia-N excretion was observed in postlarvae with increasing concentration of lead. A concentration-dependent accumulation of metal was noticed in these postlarvae. Modifications in O:N ratios of postlarvae suggest that lead accumulation might have altered utilization patterns. PMID- 11886181 TI - Detoxication mechanism of exogenous monatomic phenols in pea seedlings. AB - The conversion of exogenous monatomic phenols (O-[1-(14)C]nitrophenol, 2,4-[1 14C]dinitrophenol, and alpha-[1-14C]naphthol) in pea seedlings has been investigated. It has been found that in the pea seedlings glycosylation of these phenols does not occur, but the main pathway of their detoxication is conjugation with the low-molecular-weight peptides. Approximately 80% of phenols absorbed by seedlings form phenol-peptide conjugates. The part of exogenous monatomic phenols is irreversibly bound to proteins via quinone-protein interaction. The amino acid content of the peptides involved in the conjugation process has been established. Penetration into the plant of monatomic exogenous phenols with a high dissociation constant leads to the stimulation of peptide formation. PMID- 11886182 TI - Synergistic action of ultraviolet-B radiation and cadmium on the growth of wheat seedlings. AB - The increase in ground level UV-B radiation as a result of stratospheric ozone depletion may have major deleterious effects on crop photosynthesis and productivity. Plants are exposed to UV-B and other xenobiotics simultaneously in today's industrialized world. The present studies were undertaken to see the effect of dual stress of UV-B and Cd2+ exposure on the growth of wheat (Triticum aestivum L.) seedlings. The plants grown in 0.25, 0.5, 1.0, 2.5, and 5.0 ppm Cd2+ supplemented medium were exposed to UV-B for 30 min (2 J, 0.4 mW/cm(2)) per day. After 5 and 10 days of treatment the combined stress of UV-B and Cd2+ resulted in reduction of biomass yield, growth, and chlorophyll content and changes in protein, free amino acid, starch, and total reducing sugar content. These data support the assumption that UV-B may have a regulatory role besides damaging effects and that an increased UV-B environment will likely increase this regulatory influence of UV-B radiation. The results also indicate that the adverse effects of one stress may be modulated in the presence of other stresses. PMID- 11886183 TI - Bile metabolites of polycyclic aromatic hydrocarbons in three species of fish from the Severn Estuary. AB - Six metabolites of polycyclic aromatic hydrocarbons (PAHs) were identified and quantified from the bile of 31 common eels (Anguilla anguilla), 29 European flounders (Pleuronectes flesus), and 15 conger eels (Conger conger) collected from the Severn Estuary and Bristol Channel during 1997. The bile metabolites were deconjugated by enzymatic hydrolysis and separated by reverse-phase HPLC with fluorescence detection. The major metabolite present in all fish was 1 hydroxy pyrene (75-94% of all metabolites detected) with lower proportions of 1 hydroxy chrysene (2-15%) and 1-hydroxy phenanthrene (2-8%), and small amounts of three benzo[a]pyrene derivatives (<3%). Metabolite concentrations (normalized to biliverdin content) were significantly higher in common eels than in the other two species and tended to be higher in all species at the beginning of the year than at the end. The data confirm the importance of 1-hydroxy pyrene as the key PAH metabolite in fish bile and suggest that the common eel is an ideal species for monitoring PAHs in estuarine environments. PMID- 11886184 TI - Food concentration affects the life history response of Ceriodaphnia cf. dubia to chemicals with different mechanisms of action. AB - The effect of three chemicals with different mechanisms of action (3,4 dichloroaniline, fenoxycarb, and chlorpyrifos) on the life history response of the cladoceran Ceriodaphnia cf. dubia was examined under both limited (3 x 10(4) cells/mL) and abundant (15 x 10(4) cells/mL) food conditions. Toxicity tests were conducted at both food concentrations simultaneously for each chemical, and cladocerans were examined daily from less than 24 h old until their death. A range of life history parameters were calculated, including mean brood sizes, survival, net reproductive rate, and population growth rate. The toxicity of 3,4 dichloroaniline was not significantly affected by food concentration. However, limited food significantly decreased the toxicity of fenoxycarb, and significantly increased the toxicity of chlorpyrifos. The effect of food concentration on toxicity appears to depend on the mechanism by which the chemical exerts its toxicity and on food--chemical interactions. Possible mechanisms for the different effects of food concentration on toxicity are discussed. PMID- 11886185 TI - Juvenile sea bass liver P450, EROD induction, and erythrocytic genotoxic responses to PAH and PAH-like compounds. AB - Sea bass were exposed, for 0, 2, 4, 6 and 8 h, to 0 or 2.7 microM beta naphthoflavone (BNF), or to 0, 0.1, 0.3, 0.9, or 2.7 microM benzo[a]pyrene (B(a)P) and naphthalene (NAPH). Liver cytochrome P-450 content (P450) and liver ethoxyresorufin-O-deethylase activity (EROD) induction were determined as phase I biotransformation responses, whereas erythrocytic micronuclei (EMN) and erythrocytic nuclear abnormalities (ENA) tests were performed to assess genotoxic effects. Liver alanine aminotransferase activity and liver somatic index were determined, respectively, as liver damage and common health indicators. A significant increase in sea bass EROD activity began, respectively, at 2 and 4 h exposure to 2.7 microM B(a)P and 2.7 microM BNF, whereas NAPH failed to induce EROD activity. Maximal EROD activity was observed after 6 h exposure to 2.7 microM BNF (9-fold increase), 2.7 microM B(a)P (4-fold increase), and 2.7 microM NAPH (2-fold increase), indicating BNF as the most potent EROD inducer (BNF>B(a)P>NAPH). A significant increase in liver P450 content was observed at 6 h exposure to 2.7 microM BNF (6.5-fold increase), indicating BNF as the most potent P450 inducer. A significant P450 increase was observed at 8 h exposure only to 0.1 microM B(a)P (2-fold increase), whereas it slightly increased at 2 h exposure to 2.7 microM NAPH, within a wide variable range. The BNF, NAPH, and B(a)P genotoxic potential was demonstrated as sea bass EMN and ENA. B(a)P promoted at 2 h exposure a significant EMN (24-fold) and ENA (2.2-fold) increase, whereas NAPH exhibited similar results only at 8 h exposure. BNF also increased significantly sea bass EMN (8-fold) and ENA (1.5-fold) after 8 h. The results indicated B(a)P as the most genotoxic compound, followed by NAPH and BNF (B(a)P>NAPH>BNF). Despite the low liver P450 content and EROD activity induction by NAPH and B(a)P, their genotoxic potential was higher than that of BNF. PMID- 11886186 TI - Toxicity of 40 herbicides to the green alga Chlorella vulgaris. AB - The effects on the green alga Chlorella vulgaris of 40 herbicides in 19 different chemical structure classes and with 11 dissimilar modes of action were studied through 96-h acute toxicity tests. Experimental results indicated that the average acute toxicity of acetolactate synthase (ALS)-inhibiting herbicides to C. vulgaris was close to those of the acetyl-CoA carboxylase (ACCase)-inhibiting herbicides and the lipid synthesis-inhibiting protoporphyrinogen oxidase (Protox) inhibitor. Their acute toxicities were higher than those of the microtubule process inhibitor, mitotic process inhibitor, the glutamine synthase inhibitors, and 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSP synthase) inhibitors. The acute toxicities of auxin herbicides to C. vulgaris were the lowest among all herbicides tested and that of the photosynthesis-inhibiting herbicides was the highest. PMID- 11886187 TI - Screening for soil toxicity and mutagenicity using luminescent bacteria--a case study of the explosive 2,4,6-trinitrotoluene (TNT). AB - The presented study explored the suitability of aquatic bioassays based on the marine luminescent bacterium Vibrio fischeri as screening indicators for soil toxicity and mutagenicity. The study consists of two parts: (i) determination of the bacterial toxicity and mutagenicity of the single substance 2,4,6 trinitrotoluene (TNT) and its primary reduced metabolites using three different luminescent bacteria assays and (ii) determination of the water-extractable toxicity and mutagenicity of soil samples taken at a former production plant for TNT showing complex contamination (TNT, metabolites of TNT, PAHs, and heavy metals). Resulting data indicate TNT to be predominantly responsible for the observed biological effects of soil leachates. A strategy for soil toxicity screening based on luminescent bacteria is proposed which may especially be applicable for the case of bioremediation of TNT-contaminated soils. Potentials and restrictions of this approach to soil toxicity assessment are discussed. PMID- 11886188 TI - Multibiomarker responses in fish from the Moselle River (France). AB - The response of wild fish to pollutants was studied using two biomarkers in chub (Leuciscus cephalus) at five stations in the Moselle River (France) in 1998 and in 1999. The induction of cytochrome P450 1A was quantified by the ethoxyresorufin O-deethylase (EROD) activity in the liver and the level of DNA single-strand breaks was determined in erythrocytes using the comet assay. EROD activity was observed to be up to 10-fold induced in both males and females from the downstream stations in comparison to the fish from the upstream station. Levels of DNA damage did not parallel EROD induction. Chemical analyses did not clearly explain the responses of the studied biomarkers, confirming the great difficulty in relating chemical and biological information in the field. This study confirms the difficulty in assessing the biological effects of mixtures of pollutants and points out the usefulness of a large array of biomarkers. PMID- 11886190 TI - Quality indicators for antibiotic control programmes. AB - Antimicrobial control measures are commonly perceived to lead to an improvement in quality of prescribing, cost-effectiveness and reduction in resistance. All three outcomes have been subject to scrutiny. As proper use of resources needs to be balanced with provision of high-quality health care, the effectiveness of control measures (antibiotic policies or formularies) must be monitored. Measurement of quality by using specific indicators has been suggested as an effective measure of performance. We describe a model for evaluating core aspects of antimicrobial control programmes, aimed at improving the quality of glycopeptide prescribing by 'appropriate use guidelines'. Prioritizing indicator settings within antimicrobial control programmes is essential if limited resources are to be used most effectively. Indicator development, evaluation and feedback ought to be multi-disciplinary to ensure ownership and long-term benefit. PMID- 11886191 TI - Polymicrobial ventriculitis and evaluation of an outbreak in a surgical intensive care unit due to inadequate sterilization. AB - At the end of 1999, a case of polymicrobial ventriculitis in the Department of Neurosurgery followed by an outbreak of Serratia marcescens mediastinitis in the intensive care unit of cardiovascular surgery occurred. These nosocomial surgical infections were considered to be the result of contamination of surgical sites with inadequately sterilized instruments or theatre linen. An epidemiological survey was focused on the central sterilization unit of the hospital. The microbiological results of this survey proved that the cause of the outbreak was the use of inadequately decontaminated theatre linen. This study indicates that strict infection control measures including the control of sterilization procedures and a well-organized infection control team are necessary to prevent nosocomial surgical infections. PMID- 11886192 TI - Clinical significance of extra-pulmonary involvement of invasive aspergillosis: a retrospective autopsy-based study of 107 patients. AB - Disseminated aspergillus infection has a poor prognosis, but few reports have been published on extra-pulmonary involvement in aspergillosis. We reviewed 107 autopsy records of patients with invasive aspergillosis. Fifty-five patients had extra-pulmonary aspergillosis. Organs involved included heart, kidney, central nervous system, gastrointestinal tract, spleen, liver, thyroid gland and pancreas. Extra-pulmonary aspergillosis produces different manifestations according to involved organs. Risk factors associated with dissemination included cytotoxic chemotherapy within a month of death (P=0.0087). Lack of response to empiric or preemptive treatment of amphotericin B predicted IA dissemination (P=0.0328). To improve prognosis of IA, it is important to recognize clinical features of extra-pulmonary aspergillosis and to institute the aggressive anti fungal treatment. PMID- 11886193 TI - The effect of 10% povidone--iodine solution on contaminated bone allografts. AB - The aim of this study was to determine the efficacy of 10% povidone--iodine solution for the decontamination of bone allografts. Bone samples were prepared and tested for sterility using a femoral head removed at the time of primary hip replacement. They were contaminated by a suspension of Staphylococcus epidermidis and ground to measure the quantity of micro-organism attached to the bone. Two levels of contamination were used (1 x 10(3) vs. 1 x 10(4)CFU/mL) to check the efficiency of our method of measurement. Samples of the two groups were decontaminated with 10% povidone--iodine solution using different exposure times. Before decontamination, the count of bacteria attached to the bone was proportional to the bacterial concentration of the contaminating solution. The microbiocidal activity of 10% povidone--iodine solution was the same in both groups. The decontamination time was proportional to the bacterial concentration of the contaminating solution. The results of this preliminary study suggest that a 10% povidone--iodine solution can decontaminate inoculated bone grafts, but a sufficient time of exposure according to the level of contamination must be allowed. PMID- 11886194 TI - Burkholderia cepacia complex in cystic fibrosis and non-cystic fibrosis patients: identification of a cluster of epidemic lineages. AB - This study was performed in order to compare Burkholderia cepacia complex strains from cystic fibrosis (CF) and non-CF patients at the genomovar, genetic and epidemiological levels. A total of 92 B. cepacia respiratory tract isolates were obtained from patients attending the following CF centres: Catania and Palermo, Sicily; Gualdo Tadino, Central Italy, and Milan, Northern Italy. A total of 23 B. cepacia isolates were obtained from blood, surgical wound, and intravenous catheter sources of patients without CF, hospitalized in Catania and Varese, Northern Italy. Genomovar status identification, clonality and genetic relatedness determination, antibiotic susceptibility pattern determination and electron microscopy were performed. Transmission of infection was shown in both CF and non-CF patients by identifying clonality of responsible strains. In total 13 clones were involved in cross-transmission episodes. No outbreak was described involving both CF and non-CF patients. The present study indicates the existence of a distinct cluster of strains responsible for epidemics in CF and non-CF patients, based on their genetic relatedness, distinct from strains associated with no or negligible transmissibility. This result suggests that transmissibility is not only associated with a specific genomovar in CF patients, but also with a group of genetically related lineages in CF and non-CF patients. A key role is shown for both segregation measures and careful surveillance of infection, based on selective culture, molecular identification and epidemiological characterization of individual isolates. PMID- 11886195 TI - Recurrent Sphingomonas paucimobilis -bacteraemia associated with a multi bacterial water-borne epidemic among neutropenic patients. AB - A cluster of septicaemias due to several water-related species occurred in a haematological unit of a university hospital. In recurrent septicaemias of a leukaemic patient caused by Sphingomonas paucimobilis, genotyping of the blood isolates by use of random amplified polymorphic DNA-analysis verified the presence of two distinct S. paucimobilis strains during two of the separate episodes. A strain of S. paucimobilis identical to one of the patient's was isolated from tap water collected in the haematological unit. Thus S. paucimobilis present in blood cultures was directly linked to bacterial colonization of the hospital water system. Heterogeneous finger-printing patterns among the clinical and environmental isolates indicated the distribution of a variety of S. paucimobilis clones in the hospital environment. This link also explained the multi-microbial nature of the outbreak. PMID- 11886196 TI - Contamination of central venous catheters in immunocompromised patients: a comparison between two different types of central venous catheters. AB - Catheters impregnated with silver have been proposed as a means of reducing catheter-related infection. We therefore performed a prospective randomized study to compare a new silver-impregnated central venous catheter (CVC) with a commercially available CVC in a cohort of immunocompromised patients. We studied 157 patients of whom 97 could be analysed. The median indwelling time in the study group (SC) was 10.5 days and 11 days in the control group (CC). The incidence of contamination in the SC group was 15.6 vs 24.6 in the CC group referring to 1000 catheter days. In both groups, we found 6% of catheter-related infections according to the definitions of a published scoring system. The differences between the two groups were not significant. We conclude that the SC decrease the incidence of catheter contamination and may have a positive effect on the reduction of CVC-related infections. PMID- 11886197 TI - In situ decontamination of medical wastes using oxidative agents: a 16-month study in a polyvalent intensive care unit. AB - Over a 16-month period from September 1997 to December 1998, a prospective study was made of an on-site treatment of medical wastes in a 10-bed intensive care unit. First, the wastes were ground and then, a high concentration of ozone in air was repeatedly injected into the ground wastes. The study analysed the practical application of the system and its microbiological efficiency. Inactivation experiments were made with reference strains of Staphylococcus aureus, Enterococcus hirae, Pseudomonas aeruginosa, Escherichia coli, Mycobacterium smegmatis, Bacillus subtilis var niger, Bacillus stearothermophilus, Candida albicans and Aspergillus niger. Two thousand eight hundred treatment cycles, i.e. 84,000 grindings and 140,000 ozone injections gave a treatment capacity of 50 kg of waste per day with a good staff acceptability. All kinds of medical devices used in an intensive care unit were treated. In untreated ground wastes, the median bacterial load was 105.86 (range 10(2.35) 10(8.05)) cfu/g. After ozone treatment, bacteria and fungi were reduced by a factor of 10(5). Aero-contamination of the ward was unchanged. Computer control allowed all events to be tracked. On-site medical waste treatment appears to be an efficient alternative to the usual centralized collection and treatment. PMID- 11886198 TI - Control of nosocomial Legionnaires' disease by keeping the circulating hot water temperature above 55 degrees C: experience from a 10-year surveillance programme in a district general hospital. AB - After a nosocomial outbreak of Legionnaires' disease in a 450-bed district general hospital in 1991, the circulating hot water temperature was kept above 55 degrees C as the sole control measure. From 1991 to 2000, all cases of nosocomial pneumonia were clinically monitored and tested for Legionella pneumophila serogroup 1 by serology or urinary antigen detection. Water samples from peripheral tap sites were cultured for Legionella spp. twice a year. An infection with L. pneumophila serogroup 1 was diagnosed in four out of 366 (1.1%) patients treated for nosocomial pneumonia, representing one case per 26,000 admissions. All patients were cured without complications. L. pneumophila serogroup 1 was isolated in 30 of 251 (12%) cultured hospital water samples during the monitoring period. We conclude that control of nosocomial Legionnaires' disease in a primary referral hospital is possible by keeping the circulating hospital hot water temperature above 55 degrees C, together with careful clinical surveillance. Complete eradication of Legionella spp. from the hot water system does not seem necessary. PMID- 11886199 TI - Legionella contamination of hospital water supplies: monitoring of private healthcare facilities in Bologna, Italy. AB - The hot water supplies of 11 private healthcare facilities in the city of Bologna, Italy, were monitored for the presence of Legionella spp. Four samplings were made in each establishment over a period of one year and in total 121 samples were collected from distribution points situated near the water-boiler and inside the wards (taps and showers). Legionellae were recovered from all the water supplies in question: Legionella spp. in 86.8% of samples, L. pneumophila in 82.6% of samples. L. pneumophila was found in all the water supplies at levels averaging above 10(4)cfu/L in five health facilities and reaching a maximum concentration of 10(7)cfu/L. The only parameter to have affected the presence of legionellae was the water temperature, which was seen to be inversely correlated to the concentration of Legionella spp. Despite the high levels of contamination from L. pneumophila, no cases of nosocomial Legionnaires' disease were reported during the period of the study. PMID- 11886200 TI - Nosocomial blood-stream infection in patients with end-stage renal disease: excess length of hospital stay, extra cost and attributable mortality. AB - Patients with end-stage renal disease undergoing haemodialysis are at high risk of nosocomial blood-stream infection (BSI), but data on the associated costs in this patient population are not available. Therefore, we conducted a retrospective matched (1:2) case-control study of such patients undergoing haemodialysis from January 1998 to December 1998 in a medical centre in southern Taiwan to determine the excess length of hospital stay, attributable mortality, and the extra cost caused by nosocomial BSI. The excess length of hospital stay was 30 days for cases vs. 16 days for controls (P<0.001), the mortality rate was 26.3% for cases vs. 0 for controls (P=0.003) (attributable mortality being 26.3%), and the median of overall costs was 131,584 dollars NT for cases vs. 65,282 dollars NT for controls (P<0.001). Based on these findings, we believe that an effective programme to minimize nosocomial BSI in this patient population would greatly reduce their medical and economic burdens. PMID- 11886201 TI - A survey of rotational use of biocides in hospital pharmacy aseptic units. AB - A postal survey of biocide rotation in UK hospital pharmacy aseptic units was carried out. Seventy per cent of respondents stated that biocides were rotated, most frequently in areas outside critical work zones. High-level disinfection was employed when 'aseptic' conditions were required. Decisions on frequency of rotation were most often based on in-house validation or consultation with colleagues. Toxicity and corrosiveness were the criteria rated most important in a rotation policy. Microbiological monitoring was carried out most frequently in critical work zones but less often for handwashing. Most QC hospital pharmacists supported rotation and would prefer a standard period for all applications (monthly). Guidelines need to be clarified to assist staff in decisions regarding biocide rotation. PMID- 11886203 TI - Surgical face masks in the operating theatre: are they still necessary? PMID- 11886204 TI - Surgical face masks in the operating theatre. PMID- 11886206 TI - Burkholderia cepacia from a sink drain. PMID- 11886207 TI - Molecular epidemiology for detecting Norwalk-like viruses in clinical cases and associated environment contamination. PMID- 11886208 TI - Unusual nosocomial infection due to Pseudomonas aeruginosa. PMID- 11886209 TI - Nuclear reconstitution of plant (Orychophragmus violaceus) demembranated sperm in cell-free extracts from animal (Xenopus laevis) eggs. AB - Cell-free extracts from animal Xenopus laevis egg could induce chromatin decondensation and pronuclear formation from demembranated plant (Orychophragmus violaceus) sperm. When incubated with Xenopus egg extracts, the demembranated sperm began to swell and then gradually decondensed. The assembly of the nuclear envelope in the reconstituted nuclei was visualized by means of electron microscopy and fluorescence microscopy. Membrane vesicles fused to form the double envelope around the periphery of the decondensed chromatin. The morphology of the newly formed nuclei, with a double membrane, was similar to that of nuclei after fertilization. The electron micrograph of the whole-mount prepared nuclear matrix--lamina showed the reconstituted nucleus to be filled with a dense network. PMID- 11886210 TI - The structure of mast cell secretory granules in the blind mole rat (Spalax ehrenbergi). AB - Eosinophils, basophils, and mast cells produce and secrete active substances whose role is to attack invading parasites and protect the host. In this study we use morphometric methods to study mast cells in the blind mole rat (Spalax ehrenbergi). The subterranean and solitary way of life of this species has led to the evolutionary development of special anatomical, morphological, behavioral, and physiological adaptations. Because of its particular lifestyle, the mole rat is less exposed to parasites than other rodents. This could provide a unique model for research into the pathobiology of mast cells. The paracrystalline structure of the mast cell granule content is composed of parallel plates. Diffraction analysis of electron micrographs of thin sections of araldite embedded tissues indicated that each crystal line plate is a periodic array of parallelograms. The crystal unit cell volume is approximately 930 nm(3), suggesting that each unit cell is composed of one heparin molecule and one to three additional adsorbed proteins. Morphometric data show that characteristics of the secretory granules of mast cells of the blind mole rat resemble those of other rodents. The mast cell unit granule volume in the present study was calculated to be 0.055 microm(3), similar to that of rat peritoneal mast cells. PMID- 11886211 TI - Structural variation of tracheids in Norway spruce (Picea abies [L.] Karst.). AB - The orientation of cellulose microfibrils in the cell wall and the shape and the dimensions of the cells of earlywood of four Norway spruce (Picea abies [L.] Karst.) stems grown in Finland were studied by X-ray diffraction and optical microscopy. The average microfibril angle (MFA) decreased and the diameter of the cell increased rapidly up to rings 5-10 from the pith and remained at the same level after that. The average MFA close to the pith was over 20 degrees and decreased to about 8 degrees after ring 10 from the pith. The average diameter of the cells was 35 microm in the outer rings. The shape of the cross section of the lumen changed from circular to rectangular from the pith to the bark. The tracheid length increased also as a function of the distance from the pith. The thickness of the cell wall varied between 2.8 and 3.5 microm. Automatic cell lumen and cell wall recognition procedures were developed for the analysis of the images of the cross sections of the cells. PMID- 11886212 TI - Reaction of the skin fibroblast cytoskeleton to micromanipulation interventions. AB - Micromanipulation is a strong mechanical intervention into cellular integrity and induces large changes in the fine structure of the treated cells. Human diploid skin fibroblasts (KF1 and KF2 cell lines) were chosen as an experimental model. Special hatching needles were used for defined micromanipulation interventions (deformation of plasma membrane). Changes in cytoskeletal structures were visualized by using fluorescent and confocal microscopy. The actin cytoskeleton showed a more sensitive response to micromanipulation than microtubules. Characteristic changes in microfilaments, i.e., thickenings and knot formation, were visible in treated cells fixed immediately after micromanipulation and were the result of hatching-needle pressure on the plasma membrane as well as a reaction of actin filaments localized near the plasma membrane deformation. These direct changes and also other specific alterations in the actin filament network were detectable 14 to 16 h after treatment, but they were not observed when longer reparation intervals were used. PMID- 11886214 TI - Structural development of the mineralized tissue in the human L4 vertebral body. AB - Knowledge of the structural development of the human vertebrae from non-weight bearing before birth to weight-bearing after birth is still poor. We studied the mineralized tissue of the developing lumbar L4 vertebral body at ages 15 weeks postconception to 97 years from the tissue level (trabecular architecture) to the material level (micro- and nanostructure). Trabecular architecture was investigated by 2D histomorphometry and the material level was examined by quantitative backscattered electron imaging (for typical calcium content, CaMaxFreq) and scanning small-angle X-ray scattering (for mean mineral particle thickness). During early development, the trabecular orientation changed from a radial to a vertical/horizontal pattern. For bone area per tissue area and trabecular width in postnatal cancellous bone, the maximum was reached at adolescence (20 years), while for trabecular number the maximum was reached at childhood (approximately 1 year). CaMaxFreq was lower in early bone (approximately 21 wt%) than in mineralized cartilage (approximately 29 wt%) and adolescent bone (approximately 23 wt%). In conclusion, the changes at the tissue level were observed to continue throughout life while the development of bone at the material level (CaMaxFreq, mineral particle thickness and orientation) is essentially complete after the first years of life. CaMaxFreq and mean particle thickness increase rapidly during the first years and reach saturation. Remarkably, when these parameters are plotted versus logarithm of age, they appear linear. PMID- 11886213 TI - Crystal structure of dephospho-coenzyme A kinase from Haemophilus influenzae. AB - Dephospho-coenzyme A kinase catalyzes the final step in CoA biosynthesis, the phosphorylation of the 3'-hydroxyl group of ribose using ATP as a phosphate donor. The protein from Haemophilus influenzae was cloned and expressed, and its crystal structure was determined at 2.0-A resolution in complex with ATP. The protein molecule consists of three domains: the canonical nucleotide-binding domain with a five-stranded parallel beta-sheet, the substrate-binding alpha helical domain, and the lid domain formed by a pair of alpha-helices. The overall topology of the protein resembles the structures of nucleotide kinases. ATP binds in the P-loop in a manner observed in other kinases. The CoA-binding site is located at the interface of all three domains. The double-pocket structure of the substrate-binding site is unusual for nucleotide kinases. Amino acid residues implicated in substrate binding and catalysis have been identified. The structure analysis suggests large domain movements during the catalytic cycle. PMID- 11886215 TI - Twisted plywood pattern of collagen fibrils in teleost scales: an X-ray diffraction investigation. AB - The distribution and orientation of collagen fibrils, and apatite crystals, in the scales of a bony fish (Leuciscus cephalus) were investigated by X-ray diffraction. The small-angle diffraction patterns obtained with a microfocus scanning setup from most of the examined areas exhibit a distribution of intensity of the collagen reflections according to five preferential orientations, at 36 degrees from one another. It is suggested that the peculiar small-angle X-ray diffraction pattern is due to a plywood arrangement of collagen fibrils in successive layers parallel to the surface of the scale. The fibrils are strictly aligned in each layer and the alignment rotates by 36 degrees in successive layers, according to a discontinuous twist that generates a symmetric plywood pattern. The large spread of the wide-angle reflections does not allow one to distinguish the five directions of orientation in the intensity distribution of the 002 reflection of apatite. However, the patterns recorded from the less ordered regions of the scales display two different orientations of the 002 reflection and allow one to infer a preferential distribution of the apatite crystals with their c-axes parallel to the collagen fibrils. Although much electron microscopic evidence of plywood arrangements in calcified, as well as uncalcified, tissues has been reported, these are the very first diffraction data which unambiguously confirm the presence of these peculiar structures and suggest that this kind of investigation represents a powerful tool with which to study plywood arrangements in biological tissues. PMID- 11886216 TI - Stability of membrane proteins: relevance for the selection of appropriate methods for high-resolution structure determinations. AB - High stability is a prominent characteristic of integral membrane proteins of known atomic structure. But rather than being an intrinsic property, it may be due to a selection exerted by biochemical procedures prior to structure determination, since solubilization results in the transient exposure of membrane proteins to solution conditions. This may cause structural perturbations that interfere with 3D crystallization and hence with X-ray analysis. This problem also affects the preparation of samples for electron crystallography and NMR studies and may account for the fact that high-resolution structures of representatives of whole groups, such as transport proteins and signal transducers, have not been elucidated so far by any method. A knowledge of the proportion of labile proteins among membrane proteins, and of the kinetics of their denaturation, is therefore necessary. Establishing stability profiles, developing methods to maintain lateral pressure, or preventing contact with water (or both) should prove significant in establishing the structures of conformationally flexible proteins. PMID- 11886218 TI - Crystallization and initial crystallographic analysis of the disulfide bond isomerase DsbC in complex with the alpha domain of the electron transporter DsbD. AB - The protein disulfide bond isomerase DsbC catalyzes the rearrangement of incorrect disulfide bonds during oxidative protein folding in the periplasm of Escherichia coli. The active site cysteines of DsbC are maintained in the active reduced form by the transmembrane electron transporter DsbD. DsbD obtains electrons from the cytoplasm, transports them across the inner membrane, and passes them onto periplasmic substrates, such as DsbC. The electron transport process involves several thiol disulfide exchange reactions between different classes of thiol oxidoreductase. We were able to trap the final electron transport reaction using active site mutants yielding a stable DsbC-DsbDalpha complex. This disulfide cross-linked complex was purified to homogeneity and crystallized. Dehydration of the tetragonal crystals changed the unit cell dimensions from a approximately b = 73 A, c = 267.5 A to a = b = 68.9 A, c = 230.3 A, reducing the cell volume by 23% and the solvent content from 55 to 41%. Crystal dehydration and cryo-cooling improved the diffraction quality of the crystals from 7 to 2.3 A resolution. PMID- 11886217 TI - Crystallization and preliminary X-ray diffraction analysis of the Rab escort protein-1 in complex with Rab geranylgeranyltransferase. AB - Posttranslational prenylation of proteins is a widespread phenomenon and the majority of prenylated proteins are geranylgeranylated members of the Rab GTPase family. Geranylgeranylation is catalyzed by Rab geranylgeranyltransferase (RabGGTase) and is critical for the ability of Rab protein to mediate vesicular docking and fusion of various intracellular vesicles. RabGGTase consists of a catalytic alpha/beta heterodimer and an accessory protein termed Rab escort protein (REP-1) that delivers the newly prenylated Rab proteins to their target membrane. Mutations in the REP-1 gene in humans lead to an X-chromosome-linked defect known as choroideremia--a debilitating disease that inevitably culminates in complete blindness. Here we report in vitro assembly and purification of the stoichiometric ternary complex of RabGGTase with REP-1 stabilized by a hydrolysis resistant phosphoisoprenoid analog--farnesyl phosphonyl(methyl)phoshonate. The complex formed crystals of extended plate morphology under low ionic-strength conditions. X-ray diffraction data were collected to 2.8 A resolution at the ESRF. The crystals belong to the monoclinic space group P2(1), with unit-cell parameters a = 68.7, b = 197.7, c = 86.1 A, beta = 113.4 degrees. Preliminary structural analysis revealed the presence of one molecule in the asymmetric unit. PMID- 11886219 TI - Diversity: a driving force in curriculum development. PMID- 11886220 TI - Faking a difference: evidence-based nursing and the illusion of diversity. AB - Whilst it is desirable for the dominant paradigm (discourse) in any discipline to be seen to be encouraging diversity, I will argue in this paper that the result of real diversity is to decentralize power and strengthen the challenges to that discourse. It is therefore in the interests of the dominant discourse (if it wishes to remain dominant) to act to neutralize the power of its competitors by forcing them to conform to the rules of the dominant discourse. In this way, the illusion of diversity is maintained, but its power for radical change is greatly diluted. In deconstructing some of the most influential 'texts' (in the broader sense of the term) of evidence-based nursing, I have attempted to show that the usual way of divesting competing discourses of their power is through an assimilation into the dominant discourse in the name of diversity. However, such assimilation is, in reality, more akin to an aggressive takeover bid which maintains the rules, values and culture of the dominant discourse. For example, when qualitative bids for research funding are judged according to the rules of quantitative research, that is, according to their generalizability, sample size, and so on, the outcome is inevitably to their disadvantage. Simply by agreeing to play, any competing discourse accepts the rules of the game that are designed to deny fair competition. The rhetoric of diversity therefore masks a strategy for maintaining power which can only be challenged by (Lyotard's notion of) the philosopher, who attempts to expose the inequality inherent in it. PMID- 11886222 TI - Global network explores diversity and opportunity in nurse education. PMID- 11886225 TI - Toward a model of psychological health empowerment: implications for health care in multicultural communities. AB - This article presents a model of health empowerment from an individual psychological perspective. Building on Menon's (2001) model of psychological empowerment in organizations, psychological health empowerment is developed as a construct to capture the individual community member's feelings of empowerment with regard to health and health care. The context for health empowerment is first conceptualized as an interactive system of three elements, namely, the individual community member, health service providers, and the regulatory environment consisting of health policy and systems. The individual manages his or her own health on a daily basis, interacts with health service providers when in need of specialized medical assistance, and is affected by the 'health policy and systems' element. Perceived control, perceived competence, and goal internalization, the three facets of Menon's empowerment model, are then adapted to the health context. A scale for measuring psychological health empowerment is also proposed. The implications of this approach for health care in multicultural communities are then explored. PMID- 11886228 TI - Issues of qualification assessment for nurses in a global market. AB - Many nurses have a desire to travel and work overseas for both long- and short term periods. Some may continue postgraduate study, while others may simply wish to gain experience working in another country. Depending on the country where the nurse wishes to travel and work, the process of having qualifications assessed may be quite different, usually difficult and time consuming. There are several models of assessment currently used in various countries; from examinations for all to qualification comparability, the range is diverse. These models are very relevant given that the profession has experienced a global shortage of nurses that is unlikely to improve with the current work force predictions. Reasons for the shortages range from an aging nursing population to the profession's inability to attract young people. This has lead some countries to entice nurses from other countries to lessen their shortages. This paper discusses the issues in relation to the assessment of overseas qualified nurses applying to migrate or seek employment in Australia. Some of the issues highlighted are English language assessment, qualification assessment, competency assessment, timeliness, cost and equity. Assessment models from the global market and the impact these models have on attracting nurses to Australia are discussed. PMID- 11886231 TI - The impact of globalization and environmental change on health: challenges for nurse education. AB - The environment is an established domain of nursing knowledge, but some authors argue that the traditional perspective is too narrowly focused on the immediate environment to appreciate the relevance of the global environment. This article explores how human activities are bringing about global changes through their impact on biogeochemical cycles, land use and mobility of organisms, altering biodiversity and climate, and ultimately compromising the ecosystems services that sustain our planet. The consequences of global change for population health are examined, including the emergence of drug-resistant diseases, and the implications of climate warming and pollution for health. Addressing these issues presents a considerable challenge for nursing at all levels, in promoting sustainable policies, integrating environmental considerations into clinical practice, and in the nursing role as health educators. The greatest challenge is to education, to raise awareness of the relevance and importance of the global environment to health, and to empower nurses with sufficient understanding of the issues to apply them to practice, participate in debate and contribute to policy making that aims to reduce the burden of global changes. The extent to which the profession is prepared to diversify in response to these challenges is discussed. PMID- 11886234 TI - E-learning and educational diversity. AB - This article discusses the nature of electronic learning (E-learning) and argues for its centrality to educational diversity and the shift from teaching to learning. It is argued that E-learning is the new wave strategy that sits comfortably with other strategies developed for the 21st century. As such it challenges the traditional 'banking concept' of education, where the teacher is seen as the font of knowledge as long as students acknowledge this and are eager to absorb the teacher's vital knowledge. The article argues that E-learning should replace what Freire (1994) calls the backing concept of education, which is at odds with other 21st century approaches such as lifelong learning, open and flexible learning and the accreditation of prior learning (APL) to name only a few. In suggesting the shift from the traditional approach to E-learning, the article acknowledges issues of quality assurance and the need to maintain not only standards of achievements but also the comparability of those standards. Strategies for developing E-learning material and maintaining standards are discussed. McKey (2000) and Salmon's (2001) model of E-learning development and management are used to show how E-learning works in practise. The article then focuses on the role of E-learning as a catalyst for educational diversity, freedom to learn and equality of opportunity. While E-learning encourages diversity it paradoxically creates programmes that are more specifically tailored to the market needs than traditionally validated programmes. This is seen as very good in terms of addressing specific needs, for instance, specific knowledge and skills for a particular market. The learners or students in that particular market will feel that their specific needs are recognized and addressed, and will thus see the E-learning programme as having relevance for them. The article concludes by asserting that adequate resources, particularly learner support, will distinguish quality or good programmes from bad ones. PMID- 11886236 TI - Research in Nurse Education Today: do we meet our aims and scope? AB - All issues of Nurse Education Today between January 1996 and July 2001 were examined manually and categorized as 'research' or other forms of scholarship. A total of 356 articles were reviewed and 193 of these were considered to be some form of research. The prevalence of well known methods of data collection was noted and broad trends identified. Questionnaires, reflective diaries, Delphi surveys, focus groups and individual interviews formed the backbone of methods used in educational research. There was a marked absence of experimental work. Proportionally more UK based papers avoided inferential analysis than those from overseas. We show that 'research' in Nurse Education Today has become rather narrow. It rarely incorporates 'user' (client/patient) perspectives, and rarely (especially in the UK) uses more than one site for study. More papers which undertake comparison of nurse education between countries, which employ samples from more than one country, and which address the impact of findings from an international perspective should be sought in order to enhance this aspect of the diversity of the journal. PMID- 11886239 TI - Dephosphorylation failure of tyrosine-phosphorylated STAT1 in IFN-stimulated Sendai virus C protein-expressing cells. AB - Sendai virus C protein (SeV C) has been reported to counteract the antiviral activities of interferons (IFNs) by inhibiting the expression of IFN-stimulated gene products. In SeV C-expressing cells, formation of an active ISGF3 complex and translocation of STAT1 into the nucleus were not observed. STAT1 was continuously phosphorylated at tyrosine 701 by IFN signaling; however, its serine phosphorylation was suppressed. In addition, tyrosine-phosphorylated STAT1 grew to form abnormally huge complexes. These findings suggest that the counteraction of IFN in SeV C-expressing cells is caused by disordered phosphorylation and dephosphorylation of STAT1. PMID- 11886240 TI - Single oral immunization with replication deficient recombinant adenovirus elicits long-lived transgene-specific cellular and humoral immune responses. AB - Oral-gastric delivery of vaccines is a preferred route of immunization and is particularly relevant to the development of vaccine-vector systems. We have investigated the ability of a replication deficient (E1-deleted) adenovirus construct (RAd68), which efficiently expresses the measles virus nucleocapsid (N) protein under the control of the strong HCMV IE promoter, to elicit antibody and cytotoxic T cell (CTL) responses in mice following intragastric administration. Measles virus N protein-specific CTL memory and serum antibody responses were analyzed in a total of 140 mice at time points 2-51 weeks after immunization either with a single dose of 10(8) pfu RAd68 or with a fivefold higher dose. Of the 20 animals analyzed in the first 4-week period following low-dose immunization, 6 mounted low-level splenic CTL responses while 13 animals had CTL in the mesenteric lymph nodes. Splenic CTL responses were largely undetectable at later times. Only 23% of low-dose-immunized mice made serum antibody responses and these were generally of low magnitude and frequently of short duration. In contrast, the majority of animals immunized orally with 5 x 10(8) pfu RAd68 mounted splenic CTL responses (70%) and/or antibody responses (89%). Notably, these responses were stronger and of greater duration than those seen following immunization at the lower dose. Gut mucosal immunization with replication deficient adenoviruses is a promising approach, not only for the development of complementary measles vaccine strategies which may be required for measles virus eradication, but also generally for vaccination against other infections. PMID- 11886241 TI - T cell epitopes in coxsackievirus B4 structural proteins concentrate in regions conserved between enteroviruses. AB - The present study aimed to characterize systematically the target epitopes of T cell responses in CBV4 structural proteins. These were studied by synthesizing 86 overlapping 20-aa-long peptides covering the known sequence of CBV4 structural proteins and analyzing the proliferation responses of 18 CBV4-specific T cell lines against these peptides. Recognized peptides differed depending on the HLA DR genotype of the T cell donor. They were concentrated to the VP4 and VP2 regions as six of seven common peptide epitopes located in this region, whereas there was only one in the VP3 region and none in the VP1 region. Peptides from conserved areas were recognized more often (on average, 15% of them stimulated each T cell line) than those derived from variable areas (3%) (P < 0.0001, Fisher's exact test). Some conserved peptides inducing T cell responsiveness in most subjects were identified, a knowledge which can be useful in the development of new synthetic vaccines. PMID- 11886242 TI - A comparative analysis of the avirulence and translational transactivator functions of gene VI of Cauliflower mosaic virus. AB - The primary function associated at present with the gene VI product of Cauliflower mosaic virus (CaMV) is that of a translational transactivator (TAV). In this capacity, it alters the host translational machinery to allow reinitiation of translation of other CaMV genes on the polycistronic 35S RNA of CaMV. In addition, the gene VI protein can elicit a specific type of plant defense response called the hypersensitive response (HR) in Nicotiana edwardsonii. In this study, we have adapted the agroinfiltration technique to compare the sequences of CaMV gene VI required for TAV function and elicitation of HR. To measure the activity of the TAV, we coagroinfiltrated gene VI of CaMV strain W260 with a bicistronic GUS reporter plasmid. TAV function could be assayed 4 days postinfiltration, before the onset of HR in N. edwardsonii. Through the use of the TAV and HR assays, we could show that the TAV functions of gene VI of CaMV strains W260 and D4 were equivalent, but only W260 gene VI elicited HR. A mutational analysis of W260 gene VI showed that the structural requirements for elicitation of HR were much more stringent than those for TAV function. Small deletions from either the 5' or 3' end of W260 gene VI abolished its ability to elicit HR, although the TAV function was retained in the mutant. The TAV function could also tolerate a small insertion within gene VI; this insertion abolished the elicitor function. This study provides direct evidence that the TAV function of gene VI is separate from its role as an elicitor of HR. PMID- 11886243 TI - Differences in interferon sensitivity and biological properties of two related isolates of simian virus 5: a model for virus persistence. AB - CPI(+) and CPI(-) are two canine isolates of simian virus 5 (SV5). CPI(+) was originally isolated from the cerebrospinal fluid of a dog with temporary posterior paralysis and CPI(-) was recovered at 12 days p.i. from the brain tissue of a dog experimentally infected with CPI(+). We have previously shown that the V protein of SV5 blocks interferon (IFN) signalling by targeting STAT1 for degradation. Here we report that whilst CPI(+) targets STAT1 for degradation, CPI(-) fails to and as a consequence, CPI(+) blocks IFN signalling but CPI(-) does not. Three amino acid differences in the P/V N-terminal common domain of the V protein are responsible for the observed difference in the abilities of CPI(+) and CPI(-) to block IFN signalling. In cells persistently infected with CPI(-) the virus may become repressed in response to IFN, under which circumstances virus glycoproteins are lost from the surface of infected cells and virus nucleocapsid proteins accumulate in cytoplasmic inclusion bodies. We suggest that in vivo cells infected with IFN-resistant viruses (in which there would be continuous virus protein synthesis) may be more susceptible to killing by cytotoxic T cells than cells infected with IFN-sensitive viruses (in which virus protein synthesis was repressed), and a model of virus persistence is put forward in which there is alternating selection of IFN-resistant and IFN-sensitive viruses depending upon the state of the adaptive immune response. PMID- 11886244 TI - Quantitative measurement of fusion of HIV-1 and SIV with cultured cells using photosensitized labeling. AB - The fusion of HIV and SIV with biological membranes was studied by photosensitized activation of a hydrophobic probe, [(125)I]iodonaphthylazide ([(125)I]INA), by a fluorescent lipid which is situated in the target membrane. Photosensitized labeling of viral envelope-resident proteins occurs only upon their insertion into target membranes. Photosensitized labeling as a result of HIV-1 Env-mediated cell fusion showed the same kinetics as aqueous dye transfer. We have for the first time measured kinetics of HIV and SIV virus-cell fusion. HIV-1(MN) virions were about 10x less fusion active than SIVmne virions. SIV inactivated by aldrithiol-2 retained fusion activity similar to that seen with untreated virus. The relatively slow time course of SIV-cell fusion (t(1/2) = 19 min) indicates that the fusion events are stochastic. This feature provides a basis for understanding the mode of action of HIV/SIV entry inhibitors that target transition states. PMID- 11886245 TI - Deletion of the vpu sequences prior to the env in a simian-human immunodeficiency virus results in enhanced Env precursor synthesis but is less pathogenic for pig tailed macaques. AB - The Vpu protein of human immunodeficiency virus type 1 (HIV-1) has been reported to enhance virion release from infected cells and to down-regulate the expression of CD4 on infected cells. Previous studies have shown that Vpu and the envelope glycoprotein precursor (gp160) are translated from different reading frames of the same bicistronic messenger RNA (mRNA). In order to assess the effect of the Vpu sequences 5' to the Env open reading frame on Env biosynthesis and pathogenesis, we have constructed a deletion mutant of a molecularly cloned chimeric simian--human immunodeficiency virus (SHIV(KU-1bMC33)) in which the entire coding region of vpu upstream of env had been deleted (novpuSHIV(KU 1bMC33)). While both SHIV(KU-1bMC33) and novpuSHIV(KU-1bMC33) synthesized comparable amounts of env mRNA in infected cells, the novpuSHIV(KU-1bMC33) infected cells synthesized more Env precursor when standardized against the p57 Gag precursor protein. While more Env was synthesized than Gag in novpuSHIV(KU 1bMC33)-infected cells, pulse--chase analysis revealed that p27 Gag protein was released from infected cells with delayed kinetics, a reflection of the lack of a Vpu protein. Inoculation of novpuSHIV(KU-1bMC33) into two pig-tailed macaques resulted in no loss of circulating CD4(+) T cells. However, replicating virus could be detected in the lymphoid tissues (lymph nodes, spleen, thymus) 1 year after inoculation and the thymus of one of the macaques exhibited severe atrophy. The results of these studies indicate that the Vpu coding sequences upstream of Env may attenuate the level of Env precursor biosynthesis but significantly contribute to the pathogenesis of this SHIV in pig-tailed macaques. PMID- 11886246 TI - Reexamination of amphotropic murine leukemia virus neurovirulence: neural stem cell-mediated microglial infection fails to induce acute neurodegeneration. AB - The 4070A amphotropic murine leukemia virus (A-MuLV) has been variably reported to harbor neurovirulence determinants within its env gene. In this report we reexamined this issue by applying two approaches previously demonstrated to amplify murine leukemia virus neurovirulence. The first approach involved introducing the 4070A env gene into the background of Friend virus clone FB29 to enhance peripheral virus replication kinetics and central nervous system entry. The resulting chimeric virus, FrAmE, exhibited widespread vascular infection throughout the central nervous system (CNS); however, parenchymal infection was quite limited. Neither clinical neurological signs nor spongiform neurological changes accompanied FrAmE CNS infection. To overcome this CNS entry limitation, 4070A and FrAmE were delivered directly into the CNS via transplantation of infected C17.2 neural stem cells (NSCs). Significantly, NSC dissemination of either 4070A or FrAmE resulted in widespread, high-level amphotropic virus expression within the CNS parenchyma, including the infection of microglia, the critical target required for inducing neurodegeneration. Despite the extensive CNS infection, no associated clinical neurological signs or acute neuropathological changes were observed. Interestingly, we observed the frequent appearance of circulating polytropic (MCF) virus in the serum of amphotropic virus-infected animals. However, neither peripheral inoculation of an amphotropic/MCF virus mixture nor transplantation of NSCs expressing both amphotropic and MCF viruses induced acute clinical neurological signs or spongiform neuropathology. Thus, the results generated in this study suggest that the 4070A env gene is not inherently neurovirulent. However, the frequent appearance of endogenous MCF viruses suggests the possibility that the interactions of amphotropic viruses with endogenous retroviral elements could contribute to the development of retrovirus-induced neurodegenerative disease. PMID- 11886247 TI - Chimeric recombinant hepatitis E virus-like particles as an oral vaccine vehicle presenting foreign epitopes. AB - Many viral and bacterial pathogens establish infections through mucosal surfaces in their initial stage. However, only a few nonreplicating molecules successfully induce strong mucosal immune reaction without the addition of adjuvants by oral administration. To overcome this difficulty, we investigated whether hepatitis E virus-like particles (HEV-VLPs) could be utilized as a carrier molecule for foreign antigenic epitopes and to stimulate mucosal immunity without the need for adjuvants. To accomplish this goal, we incorporated a B cell epitope tag, consisting of 11 amino acids at the C-terminal of HEV-VLP. The chimeric VLP showed morphology similar to that of the mature HEV virion and VLP. The inserted epitope was reactive with a specific monoclonal antibody in the VLP form, suggesting that it was exposed on the surface of the VLP. After oral administration without adjuvant, this chimeric HEV induced significant levels of specific IgG and IgA to both the inserted epitope and HEV-VLP in intestinal secretions. These humoral immune responses were observed as early as 2 weeks after the first immunization. These results suggest the potential of HEV-VLP as a mucosal vaccine carrier vehicle for the presentation of antigenic epitopes through oral administration. PMID- 11886248 TI - A role for human cytomegalovirus glycoprotein O (gO) in cell fusion and a new hypervariable locus. AB - A cell fusion assay using fusion-from-without (FFWO) recombinant adenoviruses (RAds) and specific antibody showed a role in fusion modulation for glycoprotein gO, the recently identified third component of the gH/gL gCIII complex of human cytomegalovirus (HCMV). As in HCMV, RAd gO expressed multiple glycosylated species with a mature product of 125 kDa. Coexpression with gH/gL RAds showed gCIII reconstitution in the absence of other HCMV products and stabilisation by intermolecular disulfide bonds. Properties of HCMV clinical isolate, Pt, also implicated gO in cell spread. Compared to laboratory strain AD169, Pt was resistant to gH antibody plaque inhibition, but mature gH was identical. However, the gO sequences were highly divergent (20%), with further variation in laboratory strain Towne gO (34%). Thus, gO forms gCIII with gH/gL, performs in cell fusion, and is a newly identified HCMV hypervariable locus which may influence gCIII's function in mediating infection. PMID- 11886249 TI - The immediate-early protein, ICP0, is essential for the resistance of herpes simplex virus to interferon-alpha/beta. AB - Herpes simplex virus type 1 (HSV-1) is resistant to the antiviral effects of interferon (IFN)-alpha, -beta, or -gamma. The fact that ICP0(-) mutants replicate like wild-type virus in IFN-alpha/beta receptor knockout mice (Leib et al., 1999, J. Exp. Med. 189, 663) suggested that ICP0 may serve a direct role in the resistance of HSV-1 to IFN. To test this hypothesis, the effects of IFN-alpha, beta, and -gamma were compared against wild-type HSV-1 and an ICP0(-) mutant virus, 7134. In Vero cells, 7134 was more sensitive to inhibition by low doses of type I IFN (-alpha/beta) or type II IFN (-gamma) than vesicular stomatitis virus, a well-studied IFN-sensitive virus. At a concentration of 100 U/ml, IFN-alpha, beta, or -gamma reduced the efficiency of 7134 plaque formation by 120-, 560-, and 45-fold, respectively. In contrast, none of the IFNs reduced wild-type HSV-1 plaque formation by more than 3-fold. Even when Vero cells were infected with 10 pfu per cell, IFN-alpha and -beta inhibited 7134 replication by over 100-fold, but inhibition by IFN-gamma decreased to less than 10-fold. While IFN-beta efficiently inhibited 7134 replication in primary mouse kidney and SK-N-SH cells, IFN-gamma did not inhibit 7134 to a comparable extent in these cells. ICP0 provided in trans from an adenovirus vector allowed 7134 to replicate efficiently in Vero cells in the presence of IFN-alpha, -beta, or -gamma. While IFN-beta or gamma efficiently repressed the ICP0 promoter-lacZ reporter gene in 7134 (i.e., approximately 60-fold reduction in beta-galactosidase activity), ICP0 provided in trans almost completely reversed IFN-mediated repression of the lacZ gene in 7134. The results suggest that the rate of ICP0 expression in infected cells in vivo may be critical in determining whether host IFNs repress the HSV-1 genome. This concept is discussed in light of its potential relevance to the establishment of latent HSV-1 infections. PMID- 11886250 TI - Reduced antigenicity of the hepatitis B virus HBsAg protein arising as a consequence of sequence changes in the overlapping polymerase gene that are selected by lamivudine therapy. AB - The prevalence of hepatitis B virus vaccine escape mutants has increased as a consequence of the introduction of global vaccination programs. Furthermore and as a consequence of the organization of the genome of hepatitis B virus (HBV) into overlapping reading frames, the selection of polymerase mutants during long term lamivudine therapy can select viruses with changes in the overlapping S gene coding for the hepatitis B small antigen (HBsAg). We have investigated the role of lamivudine in selecting HBV mutants with antigenically altered HBsAg protein using pooled human vaccine sera in enzyme immunosorbent assays and radioimmunoassays. HBsAg proteins containing the vaccine escape mutations G145R and D144E/G145R demonstrated markedly reduced binding to anti-HBs antibody. HBsAg mutants including E164D, W196S, I195M, M198I, and E164D/I195M (corresponding to the polymerase protein changes of V519L, M550I, L526M/M550V V553I, and V519L/L526M/M550V) selected during lamivudine treatment also demonstrated reduced binding to anti-HBs antibody. These findings raise the possibility of lamivudine resistant mutants arising that possess antigenically distinct HBsAg proteins. PMID- 11886251 TI - Positional effect of deletions on viability, especially on encapsidation, of Brome mosaic virus D-RNA in barley protoplasts. AB - Brome mosaic virus (BMV), a tripartite RNA plant virus, accumulates RNA3-derived defective RNAs (D-RNAs) in which 477-500 nucleotides (nt) are deleted in the central region of the 3a protein open reading frame (ORF), after prolonged infection in barley. In the present study, six artificial D-RNAs (AD-RNAs), having deletions of the same size as the naturally occurring D-RNA but at different positions in the 3a ORF, were constructed and tested for their amplification and encapsidation in barley protoplasts by coinoculation with BMV RNA1 and 2, or RNA1, 2, and 3. Northern blot analysis of RNA accumulation in total and virion fractions showed that deletions of 492 nt in the 3'-proximal and the 5'-proximal regions of the 3a ORF decreased encapsidation efficiency of the AD-RNAs compared with that of RNA3, whereas deletions in the central region enhanced encapsidation efficiency. The present results also show that deletion positions affect competition with RNA3 in the amplification and encapsidation of AD-RNAs. PMID- 11886252 TI - The 3'-untranslated region of RNA1 as a primary determinant of temperature sensitivity of Red clover necrotic mosaic virus Canadian strain. AB - Red clover necrotic mosaic virus Canadian strain (RCNMV-Can) induces symptoms on host plants at 17 degrees C, but not at 25 degrees C. We investigated the temperature sensitivity of RCNMV-Can in Nicotiana benthamiana plants and protoplasts using infectious transcripts of genomic RNAs 1 and 2. Viral RNAs accumulated in both inoculated and noninoculated leaves at 17 degrees C, whereas no viral RNAs were detected at 25 degrees C in either inoculated or noninoculated leaves. Similar temperature sensitivity in RNA accumulation was observed in protoplasts, and no viral RNAs were detected at temperatures above 22 degrees C. These results indicate that the temperature sensitivity of RCNMV-Can occurs at an early stage of infection, including during RNA replication. Using reassortant viruses and chimeric RNAs 1 between RCNMV-Can and the RCNMV Australian strain, which accumulates viral RNAs at nonpermissive temperatures for RCNMV-Can, we demonstrated that a viral determinant for the temperature sensitivity resides in the 3'-untranslated region of RNA1. PMID- 11886253 TI - Efficient gene transfer into spleen cells of newborn mice by a replication competent retroviral vector. AB - A murine leukemia virus-derived replication-competent retroviral vector with a translational cassette for the enhanced green fluorescence protein (EGFP) was previously found to function efficiently in cell culture (Jespersen et al., 1999, Gene 239, 227-235). We here report that infection of newborn NIH Swiss mice gives rise to EGFP expression in a majority of spleen cells within the first days after infection. Among the nonadherent spleen cells, B, T, and NK lymphocytes were found to be efficiently marked by EGFP by flow cytometry analysis, whereas the adherent spleen cells were negative in most animals. Analysis at time points up to 60 days after infection reveals a decline in EGFP-positive spleen cells over time. Viremia analysis and PCR analysis of spleen DNA indicate that viruses that have lost the translation cassette predominate at later stages. The results provide a model for efficient gene delivery to spleen cells in a time window of 1 to 2 weeks after infection of newborns. Although this type of vector will not in itself be applicable in the clinic, we envision that efficient in vivo gene delivery will be valuable by analysis of differentiation and proliferation of hematopoietic cells in animal models and by development and evaluation of effectors interfering with these processes. PMID- 11886254 TI - Yeast coexpression of human papillomavirus types 6 and 16 capsid proteins. AB - The L1 and L2 capsid proteins of animal and human papillomaviruses (HPVs) can self-assemble into virus-like particles (VLPs) that closely resemble native virions. The use of different animal models shows that VLPs can be very efficient at inducing a protective immune response. However, studies with infectious HPV virions and VLPs of different HPV types indicate that the immune response is predominantly type-specific. We have generated a diploid yeast strain that coexpresses the L1 and L2 capsid proteins of both HPV-6b and HPV-16, and we have purified fully assembled VLPs banding in a cesium chloride gradient at the expected density of 1.29-1.3 mg/ml. Experimental evidence strongly indicated that the four proteins coassembled into VLPs. Western blot analysis, using anti-HPV-6 and anti-HPV-16 L1-specific monoclonal antibodies and type-specific L2 antisera, demonstrated that all four proteins copurified. Most importantly, immunoprecipitation experiments, carried out using type-specific anti-L1 monoclonals and either total yeast cell extracts or purified VLPs, confirmed the interaction and the formation of covalent disulfide bonds between the two L1 proteins. Finally, HPV-6/16 VLPs administered to mice induced conformational antibodies against both L1 protein types. These results suggest that coexpression of different capsid proteins may provide new tools for the induction of antibodies directed against multiple HPV types. PMID- 11886255 TI - Characterization of adeno-associated virus rep protein inhibition of adenovirus E2a gene expression. AB - Adeno-associated virus (AAV) replication (Rep) proteins are pleiotropic effectors of viral DNA replication, RNA transcription, and site-specific integration into chromosome 19. In addition to regulating AAV gene expression, the Rep proteins modulate expression of a variety of cellular and viral genes. In this report we investigate Rep-mediated effects on expression of the adenovirus (Ad) E2a gene and the Ad major late promoter. We have found that all four Rep proteins repress E2a expression at the protein level, with Rep40 showing the weakest repression. Mutations in the purine nucleotide binding (PNB) site weakened each of the protein's abilities to repress expression. Analysis of steady-state E2a mRNA showed that Rep proteins decreased mRNA levels, but to a lesser extent than E2a protein levels. Analysis of mRNA stability demonstrated that neither Rep78 nor Rep52 affected E2a mRNA stability, suggesting that the decrease in mRNA is due to Rep-mediated inhibition of Ad E2a transcription. To determine if Rep68 proteins could directly inhibit RNA transcription, we performed in vitro transcription assays using HeLa nuclear extracts supplemented with Rep68 and Rep68PNB. We demonstrate that Rep68, but not mutant Rep68PNB, blocked in vitro transcription of a template containing the Ad major late promoter. These results provide insight into how AAV and its encoded Rep proteins interact with Ad and provide a model system for the study of AAV and host-cell interactions. PMID- 11886256 TI - Equine herpesvirus type 1 devoid of gM and gp2 is severely impaired in virus egress but not direct cell-to-cell spread. AB - Experiments were conducted to analyze the effects of a simultaneous deletion of glycoprotein M (gM) and glycoprotein 2 (gp2) of equine herpesvirus type 1 (EHV 1). EHV-1 strain RacH was cloned as a bacterial artificial chromosome (pRacH) by homologous recombination of a mini F plasmid into the unique short region of the genome, thereby deleting gene 71 encoding gp2. Upon transfection of the pRacH DNA into rabbit kidney RK13 cells, virus plaques were visible from day 1 after transfection. The mutant RacH virus (H Delta gp2) reconstituted from pRacH lacked gene 71 and did not express gp2 as assayed by indirect immunofluorescence analysis using gp2-specific monoclonal antibodies. The H Delta gp2 virus exhibited 10-fold reduced extracellular titers and an approximately 10% reduction in mean plaque diameters when compared to parental or gp2-revertant virus. The gM open reading frame was deleted from pRacH by recE/T mediated mutagenesis in Escherichia coli. The gM-gp2 double negative virus mutant (H Delta gp2gM) did not express either of the deleted glycoproteins as demonstrated by indirect immunofluorescence analysis. The H Delta gp2gM virus exhibited a 200-fold reduction of end-point extracellular titers when compared to parental RacH virus, which could not be compensated for by growth of the mutant virus on gM-expressing cells. After restoration of the gM open reading frame, however, growth of the mutant virus was comparable to the H Delta gp2 virus. Plaque diameters of the gM gp2 double-negative mutant were reduced by only 16% when compared to that of parental RacH virus. From the results it was concluded that the simultaneous absence of gM and gp2 had an additive effect on egress but not secondary envelopment or cell-to-cell spread of EHV-1. PMID- 11886257 TI - Human immunodeficiency virus type 1 preferentially encapsidates genomic RNAs that encode Pr55(Gag): functional linkage between translation and RNA packaging. AB - Full-length retroviral RNA serves as both messenger and genomic RNA. Therefore, an unspliced RNA could play both roles: viral mRNA could be bound in cis by the same Gag polyprotein that it produced, becoming a packaged genomic RNA. To test this possibility, we used in vivo packaging experiments which coexpressed wild type NL4-3 RNA and NL4-3-based mutant RNA that, ideally, could not translate Gag. However, mutating the gag initiator produced a mutant (pNLX) that expressed a truncated Gag, Gag*, initiated at methionine 10 in the CA region (142 of Pr55(Gag)). Gag* can be rescued into virions by Gag and, as it contains the NC domain, could package RNA in cis. To eliminate NC and the CA dimerization domain, a nonsense mutation in CA at residue 99 was introduced into pNLX to produce pNLXX, which expresses an RNA that should only be packaged in trans. Cotransfection packaging experiments revealed that wild-type genomic RNA was packaged at an 8-fold greater level than NLXX RNA given equal expression of both RNAs. Experiments that varied the relative amounts of these RNAs in the cell found that the wild-type RNA was encapsidated with a packaging preference (i.e., the relative amount of this RNA in virions versus cells) of 6- to 13-fold over the NLXX RNA, showing that the NLXX RNA did not efficiently compete with NL4-3 RNA. These data suggest that the wild-type RNA's ability to express Pr55(Gag) and, by inference, actively translate Gag confers an advantage in packaging over the nearly identical NLXX RNA. In contrast, the NLX RNA competed with wild-type RNA at a 1-to-3 preference. This ratio is similar to the amounts of Gag* rescued by Gag, suggesting that the presence of Gag* assists in the encapsidation of NLX RNA. Together, our data link translation and particle formation to the packaging of viral RNA and support a model of cis packaging where nascent Gag proteins encapsidate their cognate RNA. PMID- 11886258 TI - Evidence of nucleotidyl phosphatase activity associated with core protein sigma A of avian reovirus S1133. AB - Both avian reovirus core protein sigma A purified from virus-infected cell extracts and the purified bacterially expressed protein sigma A (e sigma A) were characterized for their nucleoside triphosphate (NTP) hydrolysis activity by thin layer chromotography. Protein sigma A from both preparations has a nonspecific nucleotidyl phosphatase activity that hydrolyzes four types of NTP to their corresponding nucleoside di- and monophosphates and free phosphate. The divalent cation requirement for this activity of e sigma A was further examined by the addition of Mn(2+), Mg(2+), Ca(2+), and Zn(2+) ions. NTP hydrolysis by e sigma A was maximal when Mn(2+), Mg(2+), or Ca(2+) concentrations were 5, 4, or 1 mM, respectively. Addition of Mn(2+) or Mg(2+) stimulated the reactions up to 4- or 3 fold, respectively, higher than Ca(2+) (2.2-fold). However, Zn(2+) ion inhibited this activity of e sigma A. The results suggest that nucleotidyl phosphatase activity of e sigma A is absolutely dependent on the divalent cations Mn(2+), Mg(2+), or Ca(2+), but not Zn(2+). Similar results were obtained from the analysis of divalent cation requirements for the protein sigma A nucleotidyl phosphatase activity. Optimal pH for nucleotidyl phosphatase activity of protein sigma A from both preparations was determined using reaction mixtures buffered at different pH. The results show that the optimal activities of both proteins were similar and were achieved between pH 7.5 and 8.5. PMID- 11886259 TI - The 3C protease activity of enterovirus 71 induces human neural cell apoptosis. AB - The human glioblastoma SF268 cell line was used to investigate the induction of apoptosis by the 3C protease of enterovirus 71 (EV71). Transient expression in these cells of the wild-type 3C protein encoded by EV71 induced morphological alterations typical of apoptosis, including generation of apoptotic bodies. Degradation of cellular DNA in nucleosomes was also observed. When two of the amino acids in the catalytic motif of 3C were changed by mutagenesis, the 3C protein not only lost its proteolytic activity, but also its ability to induce apoptosis in the SF268 cells. Twenty-four hours after 3C transfection, poly(ADP ribose) polymerase, a DNA repair enzyme, was cleaved, indicating that caspases were activated by the expression of EV71 3C. The 3C-induced apoptosis was blocked by the caspase inhibitors DEVD-fmk and VAD-fmk. Our findings suggest that the proteolytic activity of 3C triggers apoptosis in the SF268 cells through a mechanism involving caspase activation and that this apoptotic pathway may play an important role in the pathogenesis of EV71 infection. PMID- 11886260 TI - Induction of secreted human immunodeficiency virus type 1 (HIV-1) resistance factors in CD4-positive T lymphocytes by attenuated HIV-1 infection. AB - This report describes induction of HIV-1 resistance and synthesis of resistance factors in immortal CD4-positive T lymphocytes. SupT1 cells were infected by NL4 3 attenuated by a defect in the vif gene through coculture with infected primary lymphocytes. Cell lines from this infection, termed R1, expressed CD4 and CXCR4, carried low levels of HIV-1 DNA, but expressed no other detectable viral products and were resistant to infection by wild-type HIV-1. Investigation of challenge infection in resistant R1 lines demonstrated entry, reverse transcription, and integration by incoming HIV-1 but no synthesis of viral RNA. By assay of marker gene expression, we found that Tat was unable to activate LTR-driven transcription in R1 lines. HIV-1-resistant R1 lines secreted soluble factors that inhibited productive infection of primary lymphocytes by several strains of HIV-1 and blocked viral RNA synthesis in newly infected cells. Resistance factors also blocked the induction of HIV-1 transcription in ACH-2 cells as assayed by viral antigen expression and Northern blot of viral RNA. Soluble factors produced by HIV-1-resistant, immortal R1 cells may form the basis of new approaches to control HIV-1 infection. PMID- 11886261 TI - HIV-1 Vpr does not inhibit CTL-mediated apoptosis of HIV-1 infected cells. AB - HIV-1 infected persons develop a robust CTL response to HIV antigens, yet HIV-1 is able to evade this host response and successfully replicate. The mechanism(s) of evasion is not completely defined but has been suggested to include resistance of infected cells to CTL-mediated apoptosis. The HIV-1 Vpr protein induces G2 arrest by indirectly inhibiting activation of cyclin B/p34cdc2 kinase. Granzyme B, the principle mediator of CTL-induced apoptosis, prematurely activates this same kinase complex. Therefore, we assessed the susceptibility of HIV-1 infected cells to CTL-mediated apoptosis to determine whether the expression of Vpr protected the infected cells from CTL-induced apoptosis. Antigen-specific CD8(+) CTL were able to induce apoptosis in HIV-1 infected cells and cells labeled with peptide corresponding to the CTL epitope with equivalent efficiency. This demonstrates that neither HIV-1 Vpr nor any other HIV protein directly inhibits CTL effector functions. Furthermore, we confirm that HIV-1 Nef is able to provide partial protection from CTL recognition of infected cells. Thus, the inability of CTL to control HIV-1 infection is likely not due to direct inhibition of CTL mediated apoptosis. PMID- 11886262 TI - Bovine herpesvirus 1 glycoprotein G is necessary for maintaining cell-to-cell junctional adherence among infected cells. AB - Glycoproteins gE and gG of bovine herpesvirus 1 (BHV-1) are involved in viral cell-to-cell transmission. We have compared the subcellular localizations of gE and gG and examined the cell-to-cell adherence of bovine kidney (MDBK) cells infected with BHV-1 mutants lacking gE or gG. In BHV-1-infected MDBK cells, gE was observed at cell junctions but did not localize at apical or basal plasma membranes. BHV-1 gG was primarily found in the cytoplasm and was also observed at boundaries among infected cells. During the infection with wild-type or gE negative BHV-1, the filamentous actin and the adherent junctional proteins accumulated at the cell junctions. In contrast, cell junctions of MDBK cells infected with gG-negative BHV-1 were loosened, and the junctional proteins and BHV-1 gE were distributed in the cytoplasm. These data indicate that BHV-1 gG facilitates viral cell-to-cell spread by maintaining the cell-to-cell junctions among the infected cells. PMID- 11886263 TI - Peripheral T-cell lymphoma in herpesvirus saimiri-infected tamarins: tumor cell lines reveal subgroup-specific differences. AB - Efficiency of lymphoma induction by herpesvirus saimiri (HVS) isolates correlates with the genetically defined viral subgroups A, B, and C. To compare subgroup specific effects, highly susceptible tamarins were infected with HVS strain A-11, B-SMHI, or C-488. All animals developed T-cell lymphomas indistinguishable with respect to clinical, pathological, and virological parameters. Ex vivo T-cell lines were established readily from the HVS C-488 animal, less efficiently in the presence of HVS A-11, and from only a single HVS B-SMHI sample. These cultivated cells revealed strain-specific biochemical characteristics. HVS A-11 strongly induced the expression of tyrosine kinase Lyn. HVS C-488 led to the activation of STAT3, which is most likely linked to the association of virus-encoded Tip with tyrosine kinase Lck. The lack of these activities in HVS B-SMHI-transformed cells may correlate with the reduced oncogenic phenotype of this virus in species other than tamarins. PMID- 11886264 TI - Enhanced immunogenicity of HPV 16 E7 fusion proteins in DNA vaccination. AB - DNA vaccination is a promising approach for inducing both humoral and cellular immune responses. For immunotherapy of HPV-16-associated diseases the E7 protein is considered a prime candidate, as it is expressed in all HPV-16-positive tumors. Unfortunately, the E7 protein is a very poor inducer of a cytotoxic T cell response, when being used as antigen in DNA vaccination. Here we demonstrate that after fusion to protein export/import signals such as the herpes simplex virus ferry protein VP22, E7 can translocate in vitro from VP22-E7-expressing cells to neighboring cells that do not carry the VP22-E7 gene. In vivo, the VP22 E7 fusion shows significantly increased efficiency in inducing a cytotoxic T-cell response. Our data suggest that the export function of VP22 plays a major role in this phenomenon, since VP22 can be replaced by classical protein export signals, without impairing the induction of the E7-specific cellular immune response. However, all E7 fusion constructs showed significantly elevated protein steady state levels, which might also account for the observed boost in immunogenicity. PMID- 11886265 TI - Hantaan virus enters cells by clathrin-dependent receptor-mediated endocytosis. AB - The cellular entry of Hantaan virus (HTN) occurs through interactions with beta(3) integrins as cellular receptors. However, the process of HTN infection following attachment to the cell surface is not well understood. Our data indicate that overexpression of a dominant-negative mutant dynamin inhibits HTN internalization and that compounds that block clathrin- but not caveolae dependent endocytosis also reduce HTN infectivity. In addition, we show that HTN colocalizes with the clathrin heavy chain but not with caveolae. At the early phase of infection HTN colocalizes with EEA-1, an early endosome marker, and later, HTN colocalizes with LAMP-1, a lysosome marker. Cells treated with lysosomotropic agents are largely resistant to infection, suggesting that a low pH-dependent step is required for HTN infection. These findings demonstrate that HTN enters cells via the clathrin-coated pit pathway and uses low-pH-dependent intracellular compartments for infectious entry. PMID- 11886266 TI - An antibody that prevents the hemagglutinin low pH fusogenic transition. AB - We have determined the structure of a complex of influenza hemagglutinin (HA) with an antibody that binds simultaneously to the membrane-distal domains of two HA monomers, effectively cross-linking them. The antibody prevents the low pH structural transition of HA that is required for its membrane fusion activity, providing evidence that a rearrangement of HA membrane-distal domains is an essential component of the transition. PMID- 11886267 TI - A replication-competent chimera of plant and animal viruses. AB - Human, animal, fungal, and plant viruses encode papain-like proteinases that function in polyprotein processing, RNA synthesis, and virus-host interactions. To compare the functional profiles of diverse papain-like proteinases, we replaced a proteinase gene of the beet yellows virus (BYV) with those derived from equine arteritis virus (EAV), foot-and-mouth disease virus (FMDV), and the fungal virus CHV1. We found that, although each of the foreign proteinases efficiently processed the viral polyprotein, only the EAV proteinase supported vigorous replication of the chimeric BYV in plant protoplasts. This result demonstrated that the proteinases of BYV and EAV, but not FMDV or CHV1, provide a function that is critical for genome replication and that is separable from polyprotein processing. Further characterization of the BYV-EAV chimera revealed that BYV proteinase is also required for virus invasion and cell-to-cell movement. Thus, the same viral protein can combine both replication-related functions shared by plant and animal viruses and specialized functions in virus host interactions. PMID- 11886268 TI - Altered cell growth and morphology in a BHK-21 cell mutant that lacks a receptor for Theiler's murine encephalomyelitis virus. AB - The receptor for Theiler's murine encephalomyelitis virus (TMEV) remains unknown. In vitro, BHK-21 cells are permissive to infection by TMEV. Selecting mutants of BHK-21 cells produced a cell line (BHKR-) resistant to infection by TMEV. Viral persistence was ruled out by immunofluorescent staining for viral antigens. BHKR- cells were nonpermissive to infection even at high multiplicities of infection. In contrast, cells were able to support one round of virus replication when transfected with infectious TMEV RNA. Binding studies indicated that TMEV was unable to attach to these cells. These data are consistent with the BHKR- cells lacking a receptor for TMEV. Interestingly, BHKR- cells were larger in size and had a significant lag in growth after subculture versus BHK-21 cells. This suggests that the TMEV receptor on BHK-21 cells could play an important role in cell growth and morphology under physiologic conditions. BHKR- cells should facilitate the search for TMEV receptors. PMID- 11886270 TI - Sequence and organization of the Mamestra configurata nucleopolyhedrovirus genome. AB - The nucleotide sequence of the genome of the nucleopolyhedrovirus (NPV) from Mamestra configurata (MacoNPV, isolate 90/2), a group II NPV, was determined and analyzed. The MacoNPV DNA genome consists of 155,060 bp and has an overall G+C content of 41.7%. Computer-assisted analysis predicted 169 open reading frames (ORFs) of 150 nucleotides or greater that showed minimal overlap. BLAST searches and comparisons with completely sequenced baculoviruses indicated that there were 66 ORFs conserved among the nine baculoviruses compared and an additional 17 ORFs were conserved among the NPVs. The gene content and gene arrangement in MacoNPV were most similar to those of SeMNPV, including two putative odv-e66 and p26 gene homologues. However, in contrast to SeMNPV, 8 ORFs with homology to baculovirus repeat ORFs (bro) and single copies of enhancin and conotoxin-like protein ORFs were found in MacoNPV. The MacoNPV genome contained four homologous regions, each with 10 to 17 repeated sequences. Each repeat was 60 to 86 nucleotides in length and contained an approximately 43-bp-long imperfect palindrome. There were 13 ORFs unique to MacoNPV, ranging from a small ORF of 196 bp to larger ORFs of up to 1047 bp, and many of these contained typical early and late baculovirus consensus promoters. PMID- 11886269 TI - Hepatitis C virus NS5A protein impairs TNF-mediated hepatic apoptosis, but not by an anti-FAS antibody, in transgenic mice. AB - Hepatitis C virus (HCV) is a major etiologic agent of chronic hepatitis worldwide and may lead to the development of hepatocellular carcinoma. However, the mechanism of development of chronic hepatitis or hepatocarcinogenesis by HCV remains unclear. In the present study, we have investigated the effect of nonstructural protein 5A (NS5A) on TNF- and Fas-mediated apoptosis in the liver of transgenic mice. For this purpose, transgenic mice were generated by targeting the HCV NS5A genomic region cloned under the control of a liver-specific apoE promoter. The transgenic animals were phenotypically similar to their normal littermates and did not exhibit a detectable histological change in the liver at 8-12 weeks of age. Intraperitoneal injection of recombinant TNF induced hepatic injury and apoptosis in normal mice. In contrast, transgenic mice expressing NS5A protein were protected against hepatic apoptosis after injection of TNF. However, injection of anti-Fas antibody into transgenic mice did not significantly influence hepatic apoptosis compared to the normal littermates. These results suggested distinct effects of TNF and anti-Fas antibody in transgenic mice expressing NS5A. We subsequently investigated the effect of NS5A in signaling pathways involved in these two cytokine-mediated apoptosis. A physical association between NS5A and TRADD was observed by pull-down assay, coimmunoprecipitation, and colocalization experiments. Furthermore, NS5A prevented the association between TRADD and FADD and blocked TRADD-mediated NF kappaB activation. Together, our results suggest that NS5A impairs TNF-mediated apoptosis by interfering upstream of the signal transduction pathway and may play a role in HCV-mediated pathogenesis. PMID- 11886271 TI - Nontemplated base addition by HIV-1 RT can induce nonspecific strand transfer in vitro. AB - After minus-strand strong-stop DNA (-sssDNA) synthesis, the RNA template is degraded by the RNase H activity of reverse transcriptase (RT), generating a single-stranded DNA. The genomes of some retroviruses contain sequences that could lead to self-priming of their minus signsssDNA. Self-priming was prevented by annealing a DNA oligonucleotide to the 3' end of model DNAs that corresponded to the 3' ends of the -sssDNAs (-R ssDNA) from human immunodeficiency virus type 1 (HIV-1), type 2 (HIV-2), and human T-cell leukemia virus type 1 (HTLV-1) but nonspecific strand transfer to ssDNA molecules in solution was induced in vitro (Golinelli and Hughes, 2001). This nonspecific strand transfer involved the addition of a nontemplated base to the 3' end of -R ssDNAs that was part of a blunt-ended duplex. In the case of HIV-2 -R ssDNA, A and C were added more efficiently than G and T. Strand transfer to ssDNA in solution occurred only if the nontemplated base could form a basepair with the last base at the 3' end of the ssDNA. If there was a mismatch, strand transfer did not occur. There was no detectable strand transfer to internal sites in the target ssDNA except to the second position from the 3' end of the DNA acceptor when the sequences at the 3' ends of the two DNAs allowed the formation of two basepairs. The nontemplated base addition and the one-basepair strand transfer were both affected by the salt concentration in the reaction, the nature of the reverse transcriptase (HIV-1 versus Moloney murine leukemia virus), and the sequence at the 3' end of -R ssDNA. NC reduced the efficiency of nonspecific strand transfer in vitro, suggesting that NC may have a role in reducing nonspecific strand transfer in vivo. PMID- 11886272 TI - In vivo tissue-specific regulation of the human papillomavirus type 18 early promoter by estrogen, progesterone, and their antagonists. AB - Human papillomavirus type 18 is a causative agent of epithelial cancers in the uterine cervix. We show here that estrogen and progesterone activate beta galactosidase expression from the early promoter of this virus in the genital epithelia of transgenic mice. Ovariectomy caused suppression of transgene expression exclusively in vagina and cervix epithelia. Beta-galactosidase expression could be restored in ovariectomized females by administration of estrogen, alone or in combination with progesterone. Further, rescue of transgene expression was inhibited by the estrogen antagonist tamoxifen and the anti progesterone RU486, suggesting that this was a specific effect. PMID- 11886273 TI - Identification of a minimal HIV-1 gag domain sufficient for self-association. AB - Gag polyprotein precursors play an essential role in the assembly of the HIV particle by polymerizing into a spherical shell at the plasma membrane. In order to define the domains within Gag responsible for this homotypic interaction, we have coupled the technology of the yeast two-hybrid system with the technology of a gene-based, semirandom library. By this method, we have identified a minimal region of Gag capable of efficient self-interaction. This region consists of the N-terminal portion of the nucleocapsid protein (NC), including the first zinc finger and the previously described interaction, or I, domain. In parallel with this randomized approach, individual HIV Gag domains, and combinations of these domains, were tested for potential homotypic and heterotypic interactions in the yeast two-hybrid system. Consistent with the results from the semirandom library screen, only combinations of species containing NC were strongly interacting. PMID- 11886274 TI - Spontaneous mobilization of integrated recombinant adenoassociated virus in a cell culture model of virus latency. AB - A cell line containing integrated recombinant adenoassociated virus (AAV) was investigated for spontaneous mobilization of vector sequence. Detection of these rare events was facilitated by using a vector design that allowed the circular rescue product (cAAV) to be individually scored by bacterial transformation. Restriction and sequence analysis of captured clones revealed five highly ordered classes of cAAV, each of which contained a defined segment of the integrated vector locus. A common feature of all cAAV classes was the presence of a modified inverted terminal repeat that joined the ends of the liberated sequence. Assembly of extrachromosomal vector genomes was accompanied by deletions in the integration locus that could be mapped to one of the five cAAV classes, suggesting an excision-type mechanism. We propose that the spontaneous deletion and mobilization of vector sequence from the recombinant adenoassociated virus (rAAV) integration locus is mediated by a recombination event between the inverted terminal repeats that define the boundaries of the individual genome subunits. PMID- 11886275 TI - SP1 and AP-1 elements direct chromatin remodeling in SV40 chromosomes during the first 6 hours of infection. AB - To identify the SV40 regulatory sequences responsible for the chromatin remodeling associated with early transcription, SV40 chromosomes containing potential remodeling sequences inserted adjacent to a reporter region were isolated at various times within the first 6 h of infection and analyzed by a combination of restriction endonuclease digestion and competitive PCR amplification. The sequences analyzed included the early domain, the enhancer, the late domain, the early phasing element, the AP-1 element, two tandem copies of the SP1 element, and the AP-4 element. From 30 min to 3 h postinfection only the enhancer, the AP-1 element, and the two tandem copies of the SP1 element caused a change in nuclease sensitivity consistent with chromatin remodeling. These results suggest that the changes in chromatin structure seen in the promoter during activation of early transcription are most likely a result of remodeling by the AP-1 and/or SP1. PMID- 11886276 TI - Spliced and prematurely polyadenylated Jaagsiekte sheep retrovirus-specific RNAs from infected or transfected cells. AB - Jaagsiekte sheep retrovirus (JSRV) is the etiologic agent of a contagious lung cancer of sheep, ovine pulmonary adenocarcinoma (OPA). In this study, we characterized the virus-specific RNAs in 293T cells transiently transfected with a human cytomegalovirus promoter-driven JSRV expression plasmid, in productively infected OHH1.LU deer lung cells, and in OPA tumors from field isolates. Typical unspliced (presumably for gag, pro, and pol) and singly spliced env mRNAs were detected. In addition, six other virus-specific RNAs were detected that resulted from the use of alternate splice acceptor sites and two premature polyadenylation sites (located in gag and in env). The orf-x gene of the virus appears to be expressed from two singly spliced subgenomic mRNAs of 3.2 kb that would encode an independent orf-x protein of 179 amino acids. In addition, the results suggested that there may also be an internal promoter for orf-x. PMID- 11886277 TI - Splicing inhibition at the level of spliceosome assembly in the presence of herpes simplex virus protein ICP27. AB - Herpes simplex virus (HSV) immediate-early protein ICP27 is a multifunctional regulator of viral and cellular gene expression. It has previously been shown that ICP27 directly or indirectly modulates several posttranscriptional processes, such as pre-mRNA splicing and polyadenylation. We show here that pre mRNA splicing is inhibited in nuclear extracts prepared from cells in which ICP27 has been transiently expressed. Our results show that splicing inhibition in ICP27 extracts is manifested at early stages of the splicing process. Furthermore, our results suggest that an enzymatic activity in ICP27-containing extracts causes the splicing inhibition. PMID- 11886278 TI - Identification and characterization of novel murine cytomegalovirus M112-113 (e1) gene products. AB - The human cytomegalovirus (HCMV) UL112-113 gene products play important roles in viral DNA replication and transcriptional regulation. In this report, we characterize two novel transcripts originating from the homologous M112-113 (e1) region of the murine cytomegalovirus (MCMV) genome. These transcripts of 2.0 and 2.4 kb represent alternatively spliced products of the e1 gene region. Analysis of the e1 proteins demonstrates the presence of a previously unidentified 87-kDa protein that is likely encoded by the 2.4-kb transcript. All four protein products derived from the e1 gene region are expressed with early kinetics, are coordinately regulated, and localize predominantly to the nucleus of MCMV infected cells. The expression pattern and localization of the e1 proteins show significant similarity to those of the HCMV UL112-113 proteins, signifying that MCMV e1 will serve as a useful model for assessing the role of this early gene region during viral infection. PMID- 11886279 TI - Simian immunodeficiency viruses with defective nef genes show increased susceptibility to the noncytotoxic antiviral activity of CD8+ lymphocytes. AB - The noncytotoxic soluble factor produced by CD8+ T cells inhibits replication of HIV and SIV in vitro and is thought to play a crucial role in combatting infection in vivo. We determined the effect of human CD8+ lymphocytes on the in vitro replication potential of both wild-type and nef-defective mutants of the simian immunodeficiency virus SIVmac251. Although replication of wild-type SIVmac251 in unstimulated human PBMC supplemented with IL-2 was unaffected by the presence of CD8+ T cells, the nef mutants were susceptible to the inhibitory effects. The effect of exogenous IL-2 depended upon the culture conditions: (i) in nonstimulated human PBMC depleted of CD8+ T cells, addition of IL-2 had a positive effect on the growth of the nef-defective viruses; (ii) in total human PBMC, IL-2 appeared to reinforce the CD8+ T-cell-dependent inhibition of the same mutant viruses. This strongly suggests that IL-2 stimulates the noncytotoxic anti HIV/SIV response of CD8+ cells present in PBMC cultures. PHA stimulation of unfractionated human PBMC overrode the suppression of viral replication by CD8+ T cells. Depletion of activated T cells expressing the IL-2 receptor alpha-chain (CD25+ T cells), present in small amounts in these primary T cell cultures, dramatically reduced viral replication, indicating that the depleted cell population harbors the target cells permissive for viral replication. Furthermore, using neutralizing antibodies we could show that inhibition by the beta-chemokines MIP-1alpha, MIP-1beta, and RANTES and the inhibitory effect of CD8+ lymphocytes on nef mutant SIVmac viruses are harbored on different levels. PMID- 11886281 TI - Increasing rates of reaction: microwave-assisted organic synthesis for combinatorial chemistry. PMID- 11886280 TI - The spike but not the hemagglutinin/esterase protein of bovine coronavirus is necessary and sufficient for viral infection. AB - The spike (S) and hemagglutinin/esterase (HE) of bovine coronavirus (BCV) are the two envelope proteins that recognize the same receptor-determinant of 9-O acetylneuraminic acid on host cells. However, the precise and relative roles of the two proteins in BCV infectivity remain elusive. To unequivocally determine their roles in viral cytopathogenicity, we developed a system in which phenotypically chimeric viruses were generated by infecting a closely related mouse hepatitis virus (MHV) in cells that stably express an individual BCV protein (S or HE). The chimeric viruses were then used to infect human rectal tumor (HRT)-18 cells that are permissive to BCV but are nonsusceptible to MHV. Using this approach, we found that the chimeric virus containing the BCV S protein on the virion surface entered and replicated in HRT-18 cells; this was specifically blocked by prior treatment of the virus with a neutralizing antibody specific to the BCV S protein, indicating that the BCV S protein is responsible for initiating chimeric virus infection. In contrast, chimeric viruses that contain biologically active and functional BCV HE protein on the surface failed to enter HRT-18 cells, indicating that the BCV HE protein alone is not sufficient for BCV infection. Taken together, these results demonstrate that the S protein but not the HE protein of BCV is necessary and sufficient for infection of the chimeric viruses in HRT-18 cells, suggesting that BCV likely uses the S protein as a primary vehicle to infect permissive cells. PMID- 11886282 TI - Solid-phase synthesis of unsaturated 3-substituted piperazine-2,5-diones. PMID- 11886284 TI - Dendritic aliphatic polyethers as high-loading soluble supports for carbonyl compounds and parallel membrane separation techniques. AB - This paper describes the use of dendritic polyglycerol as a new high-loading polymeric support. The soluble polyether skeleton allows the parallel synthesis of small libraries on a large scale (1-5 mmol). Purification of polymer-bound products is easily achieved by a parallel dialysis apparatus, which was developed to separate up to 12 reaction mixtures simultaneously. The terminal 1,2-diol groups of polyglycerol (loading capacity: 4.1 mmol diol/g) can be directly coupled with carbonyl compounds without additional linker groups. At the same time the polyglycerol support acts as a polymeric ketal protecting group. The coupling of the carbonyl compounds occurs in high yields, and effective loading capacities of up to 3.5 mmol of ketone/g can be reached. The obtained polymeric acetals can easily be characterized by standard analytical techniques, such as NMR, IR, UV, and SEC. The versatility of this new polymeric support for solution phase organic synthesis is demonstrated by two efficient polymer-supported syntheses: nucleophilic substitutions of gamma-chloroketones with amines and Suzuki-coupling on p-bromobenzaldehyde. The acid-catalyzed acetal cleavage with a solid-phase acidic ion-exchange resin in methanol demonstrates the orthogonal use of these soluble polymeric supports with conventional solid-phase reagents. Cleavage of products occurs in high yields, and almost complete recovery (>95%) of the polyglycerol support has been demonstrated after phase separation or ultrafiltration. PMID- 11886283 TI - In situ generation of carbon monoxide from solid molybdenum hexacarbonyl. A convenient and fast route to palladium-catalyzed carbonylation reactions. PMID- 11886285 TI - Color test for the detection of resin-bound aldehyde in solid-phase combinatorial synthesis. AB - We report the development of a sensitive and specific color test for the detection of the presence of resin-bound aldehyde groups using 4-amino-3 hydrazino-5-mercapto-1,2,4-triazole (Purpald). Aldehyde resin turns dark-brown to purple after a 5 min reaction followed by a 10 min air oxidation period. Resins that possess other functional groups (i.e., ketone, ester, amide, alcohol, and carboxylic acid) do not change color under the same conditions. The detection limit is 20 micromol/g for polystyrene-based aldehyde resins. PMID- 11886286 TI - Templates for exploratory library preparation. Derivatization of a functionalized spirocyclic 3,6-dihydro-2H-pyran formed by ring-closing metathesis reaction. AB - The preparation of a novel spirocyclic template from tert-butoxycarbonyl-4 piperidone is reported. The synthesis of N-(tert-butoxycarbonyl)-1-oxa-9-aza spiro[5.5]undec-3-ene (4) for exploratory library generation involves ketone allylation, etherification, and ring-closing metathesis (RCM) reactions. Epoxidation of the alkene formed in the RCM followed by addition of volatile amines to the epoxides led rapidly to an exploratory library of structurally novel spirocyclic amino alcohols. The addition of amines to epoxides derived from 4 was determined to occur primarily at C3. PMID- 11886287 TI - "Meshed-Bag Gathered-Bunch" method for solid-phase synthesis of small molecular diverse compounds. AB - A new "Meshed-Bag Gathered-Bunch" technology for the solid-phase synthesis of chemical libraries was developed. Using such technology, we synthesized muramyl dipeptide mimetics including derivatives at the N- and C-terminus, cyclic muramyl dipeptide mimetics, muramyl dipeptide and Tuftsin's analogue conjugates. The advantages of such a method include ease of manufacture, low unit cost of production, the physical encoding method, and the compatibility with both parallel and "split-mix" approaches. PMID- 11886288 TI - Development of a system to evaluate compound identity, purity, and concentration in a single experiment and its application in quality assessment of combinatorial libraries and screening hits. AB - The development and use of a new assay system for the simultaneous determination of identity, purity, and concentration of sample components from combinatorial libraries produced by parallel synthesis are described. The system makes use of high-performance liquid chromatography with UV/vis photodiode array (PDA), evaporative light scattering (ELSD), chemiluminescent nitrogen (CLND), and time of-flight mass spectrometer (TOFMS) detectors (HPLC-PDA-ELSD-CLND-TOFMS). Although these detectors have previously been utilized separately for the analysis of combinatorial chemistry libraries, the use of TOFMS along with CLND provides a synergistic combination enabling target and side-product structures to be identified and their concentrations and purities determined in a single experiment from a solution containing microgram levels of material. The CLND was found to give a linear response based on the number of moles of nitrogen present. Therefore, if the number of nitrogens per molecule is known, the concentration of each nitrogen-containing sample component may be determined utilizing an unrelated co-injected standard. A molecular formula for an impurity may often be calculated from the exact mass determined by the TOFMS and knowledge of the chemistry involved. Thus, if the sample components contain nitrogen, the concentration of every identified HPLC peak may be determined even in the absence of primary standards. This combination of detectors enabled the characterization of both target compounds and byproducts in combinatorial libraries, allowing the optimization of library synthetic procedures. This system was also used to survey the quality of libraries, enabling the selection of the best libraries for screening. This method also facilitated the characterization of samples from combinatorial libraries found as hits in high-throughput screening to establish the potency of the leads based on their actual concentration. In addition, concentrations and potencies of impurities were determined after identification of their structures, utilizing exact mass data, determination of charge states, and knowledge of the synthetic chemistry. PMID- 11886289 TI - Solid-phase synthesis of thalidomide and its analogues. AB - A novel solid-phase synthesis of thalidomide and its metabolites and analogues is described. The synthetic strategy involves the coupling of hydroxymethyl polystyrene with phthalic anhydride to form the resin-linked acid. The acid is then reacted with primary amines followed by acid or base treatment to form thalidomide and its analogues with either open or closed phthalimide rings. Most of the analogues are synthesized with high yields (40.3-98.1% in three steps) and purities (92.3-98.9%). PMID- 11886291 TI - A method for the parallel synthesis of multiply substituted oxazolidinones. AB - There are many examples of both naturally occurring and synthetic molecules containing a 2-oxazolidinone ring that have significant biological activity. The oxazolidinone ring potentially has three sites for attachment of diversity elements. A synthesis that can provide for inclusion of diversity elements at all three positions should be a powerful method for the preparation of oxazolidinone libraries. In this paper we present the preparation of a 3 x 3 x 3 array yielding 27 different products with minimal workup, high yields, and, most importantly, high purity. Using an intramolecular acylnitrene-mediated aziridination reaction, we have prepared (triphenylmethoxymethyl)-3-oxa-1-azabicyclo[3.1.0]hexan-2-one as a starting material for library generation. The first substitution involves opening the aziridine ring of the azabicyclo[3.1.0]hexane ring system using a Grignard reagent. The nitrogen of the oxazolidinone is now ready for substitution via alkylation or arylation. Removing the trityl protecting group via esterification under mildly acidic conditions accomplishes the final substitution. PMID- 11886290 TI - Rapid parallel synthesis of polymer-bound enones utilizing microwave-assisted solid-phase chemistry. AB - A microwave-assisted parallel solid-phase synthesis of a collection of 21 polymer bound enones has been developed. The two-step protocol involves initial high speed acetoacetylation of polystyrene Wang resin with a selection of seven common beta-ketoesters. When microwave flash heating at 170 degreesC was employed, complete conversions were achieved within 1-10 min, a significant improvement over the conventional thermal method, which takes several hours for completion. Significant rate enhancements were also observed for the subsequent microwave heated Knoevenagel condensations with a second set of 13 different aldehydes. Reaction times were reduced to 30-60 min at 125 degreesC in the microwave protocol compared to 1-2 days using conventional thermal conditions. Kinetic comparison studies indicate that the observed rate enhancements can be attributed to the rapid direct heating of the solvent (1,2-dichlorobenzene) by microwaves rather than to any specific microwave effect. All reactions have been carried out in commercially available parallel reactors with on-line temperature measurement, designed specifically for use in multimode microwave cavities. PMID- 11886292 TI - Solid-phase synthesis of trisubsituted guanidines. AB - The solid-phase library synthesis of trisubstituted guanidines was accomplished. Amines were loaded onto the 4-formyl-3,5-dimethoxyphenoxymethyl linker via reductive amination. Subsequent acylation with Fmoc-4-aminomethylbenzoic acid followed by Fmoc deprotection gave solid-supported primary amines. Alternatively, sulfonylation of resin-bound secondary amines with 4-cyanobenzenesulfonyl chloride followed by borane reduction also gave solid-supported primary amines. Both resins were acylated with isocyanates to furnish solid-supported ureas. Dehydration of ureas with p-toluenesulfonyl chloride in pyridine gave solid supported carbodiimides. Nucleophilic addition of amines to the carbodiimide bond followed by cleavage off the solid support gave trisubstituted guanidines. PMID- 11886293 TI - Solid-phase synthesis of linear ureas tethered to hydantoins and thiohydantoins. AB - An efficient method for the solid-phase synthesis of hydantoins and thiohydantoins tethered to ureas, starting from a resin-bound amino acid, is presented. Following reduction of the amide with borane-THF, a second amino acid was selectively coupled to the primary amine followed by treatment of the secondary amine by an isocyanate to generate the corresponding urea. Hydantoin and thiohydantoin formation was achieved through the use of carbonyldiimidazole and thiocarbonyldiimidazole, respectively. Cleavage from the solid support using hydrogen fluoride, followed by extraction and lyophilization, provided the desired urea-linked heterocyclic compounds in good yield and high purity. PMID- 11886295 TI - Resin-capture and release strategy toward combinatorial libraries of 2,6,9 substituted purines. AB - A resin-capture and release strategy for making combinatorial 2,6,9 trisubstituted purine libraries is demonstrated by capturing N9-derivatized purines at the C6 position with a thio-modified polymer. The C2 fluoro group is subsequently substituted with primary and secondary amines followed by thioether oxidation and release by C6 substitution with amines and anilines. This approach complements a previously reported strategy where a 6-phenylsulfenylpurine scaffold was captured at the C2 position with a resin-bound amine.(3) PMID- 11886296 TI - A wake-up call. PMID- 11886294 TI - N-arylation of primary and secondary aliphatic amines on solid supports. AB - A general and mild method for the N-arylation of primary and secondary aliphatic amines is reported. Copper acetate, triethylamine mediated C/N cross-coupling reaction of arylboronic acids at room temperature to solid-supported primary and secondary amines gave good to excellent yields of the desired N-arylated products. PMID- 11886298 TI - Postoperative nausea and vomiting--can it be eliminated? PMID- 11886299 TI - Sequencing of the malaria genome opens door to vaccines and new drugs. PMID- 11886300 TI - News about neuroprotectants for the treatment of stroke. PMID- 11886305 TI - Efficacy of intranasal corticosteroids for acute sinusitis. PMID- 11886307 TI - Does time of day of hemodialysis affect survival? PMID- 11886308 TI - Does time of day of hemodialysis affect survival? PMID- 11886309 TI - Does time of day of hemodialysis affect survival? PMID- 11886310 TI - Does time of day of hemodialysis affect survival? PMID- 11886311 TI - Does time of day of hemodialysis affect survival? PMID- 11886313 TI - Inappropriate prescribing for elderly patients. PMID- 11886315 TI - Sexual orientation and suicide risk among teenagers. PMID- 11886317 TI - Circadian variability in hemorrhagic stroke. PMID- 11886318 TI - Improving quality of care for acute myocardial infarction: The Guidelines Applied in Practice (GAP) Initiative. AB - CONTEXT: Quality of care of patients with acute myocardial infarction (AMI) has received intense attention. However, it is unknown if a structured initiative for improving care of patients with AMI can be effectively implemented at a wide variety of hospitals. OBJECTIVE: To measure the effects of a quality improvement project on adherence to evidence-based therapies for patients with AMI. DESIGN AND SETTING: The Guidelines Applied in Practice (GAP) quality improvement project, which consisted of baseline measurement, implementation of improvement strategies, and remeasurement, in 10 acute-care hospitals in southeast Michigan. PATIENTS: A random sample of Medicare and non-Medicare patients at baseline (July 1998--June 1999; n = 735) and following intervention (September 1--December 15, 2000; n = 914) admitted at the 10 study centers for treatment of confirmed AMI. A random sample of Medicare patients at baseline (January--December 1998; n = 513) and at remeasurement (March--August 2001; n = 388) admitted to 11 hospitals that volunteered, but were not selected, served as a control group. INTERVENTION: The GAP project consisted of a kickoff presentation; creation of customized, guideline-oriented tools designed to facilitate adherence to key quality indicators; identification and assignment of local physician and nurse opinion leaders; grand rounds site visits; and premeasurement and postmeasurement of quality indicators. MAIN OUTCOME MEASURES: Differences in adherence to quality indicators (use of aspirin, beta-blockers, and angiotensin-converting enzyme [ACE] inhibitors at discharge; time to reperfusion; smoking cessation and diet counseling; and cholesterol assessment and treatment) in ideal patients, compared between baseline and postintervention samples and among Medicare patients in GAP hospitals and the control group. RESULTS: Increases in adherence to key treatments were seen in the administration of aspirin (81% vs 87%; P =.02) and beta-blockers (65% vs 74%; P =.04) on admission and use of aspirin (84% vs 92%; P =.002) and smoking cessation counseling (53% vs 65%; P =.02) at discharge. For most of the other indicators, nonsignificant but favorable trends toward improvement in adherence to treatment goals were observed. Compared with the control group, Medicare patients in GAP hospitals showed a significant increase in the use of aspirin at discharge (5% vs 10%; P<.001). Use of aspirin on admission, ACE inhibitors at discharge, and documentation of smoking cessation also showed a trend for greater improvement among GAP hospitals compared with control hospitals, although none of these were statistically significant. Evidence of tool use noted during chart review was associated with a very high level of adherence to most quality indicators. CONCLUSIONS: Implementation of guideline-based tools for AMI may facilitate quality improvement among a variety of institutions, patients, and caregivers. This initial project provides a foundation for future initiatives aimed at quality improvement. PMID- 11886319 TI - Evaluation of a consumer-oriented internet health care report card: the risk of quality ratings based on mortality data. AB - CONTEXT: Health care "report cards" have attracted significant consumer interest, particularly publicly available Internet health care quality rating systems. However, the ability of these ratings to discriminate between hospitals is not known. OBJECTIVE: To determine whether hospital ratings for acute myocardial infarction (AMI) mortality from a prominent Internet hospital rating system accurately discriminate between hospitals' performance based on process of care and outcomes. DESIGN, SETTING, AND PATIENTS: Data from the Cooperative Cardiovascular Project, a retrospective systematic medical record review of 141 914 Medicare fee-for-service beneficiaries 65 years or older hospitalized with AMI at 3363 US acute care hospitals during a 4- to 8-month period between January 1994 and February 1996 were compared with ratings obtained from HealthGrades.com (1-star: worse outcomes than predicted, 5-star: better outcomes than predicted) based on 1994-1997 Medicare data. MAIN OUTCOME MEASURES: Quality indicators of AMI care, including use of acute reperfusion therapy, aspirin, beta-blockers, angiotensin-converting enzyme inhibitors; 30-day mortality. RESULTS: Patients treated at higher-rated hospitals were significantly more likely to receive aspirin (admission: 75.4% 5-star vs 66.4% 1-star, P for trend =.001; discharge: 79.7% 5-star vs 68.0% 1-star, P =.001) and beta-blockers (admission: 54.8% 5-star vs 35.7% 1-star, P =.001; discharge: 63.3% 5-star vs 52.1% 1-star, P =.001), but not angiotensin-converting enzyme inhibitors (59.6% 5-star vs 57.4% 1-star, P =.40). Acute reperfusion therapy rates were highest for patients treated at 2 star hospitals (60.6%) and lowest for 5-star hospitals (53.6% 5-star, P =.008). Risk-standardized 30-day mortality rates were lower for patients treated at higher-rated than lower-rated hospitals (21.9% 1-star vs 15.9% 5-star, P =.001). However, there was marked heterogeneity within rating groups and substantial overlap of individual hospitals across rating strata for mortality and process of care; only 3.1% of comparisons between 1-star and 5-star hospitals had statistically lower risk-standardized 30-day mortality rates in 5-star hospitals. Similar findings were observed in comparisons of 30-day mortality rates between individual hospitals in all other rating groups and when comparisons were restricted to hospitals with a minimum of 30 cases during the study period. CONCLUSION: Hospital ratings published by a prominent Internet health care quality rating system identified groups of hospitals that, in the aggregate, differed in their quality of care and outcomes. However, the ratings poorly discriminated between any 2 individual hospitals' process of care or mortality rates during the study period. Limitations in discrimination may undermine the value of health care quality ratings for patients or payers and may lead to misperceptions of hospitals' performance. PMID- 11886320 TI - Racial disparities in the quality of care for enrollees in medicare managed care. AB - CONTEXT: Substantial racial disparities in the use of some health services exist; however, much less is known about racial disparities in the quality of care. OBJECTIVE: To assess racial disparities in the quality of care for enrollees in Medicare managed care health plans. DESIGN AND SETTING: Observational study, using the 1998 Health Plan Employer Data and Information Set (HEDIS), which summarized performance in calendar year 1997 for 4 measures of quality of care (breast cancer screening, eye examinations for patients with diabetes, beta blocker use after myocardial infarction, and follow-up after hospitalization for mental illness). PARTICIPANTS: A total of 305 574 (7.7%) beneficiaries who were enrolled in Medicare managed care health plans had data for at least 1 of the 4 HEDIS measures and were aged 65 years or older. MAIN OUTCOME MEASURES: Rates of breast cancer screening, eye examinations for patients with diabetes, beta blocker use after myocardial infarction, and follow-up after hospitalization for mental illness. RESULTS: Blacks were less likely than whites to receive breast cancer screening (62.9% vs 70.9%; P<.001), eye examinations for patients with diabetes (43.6% vs 50.4%; P =.02), beta-blocker medication after myocardial infarction (64.1% vs 73.8%; P<.005), and follow-up after hospitalization for mental illness (33.2 vs 54.0%; P<.001). After adjustment for potential confounding factors, racial disparities were still statistically significant for eye examinations for patients with diabetes, beta-blocker use after myocardial infarction, and follow-up after hospitalization for mental illness. CONCLUSION: Among Medicare beneficiaries enrolled in managed care health plans, blacks received poorer quality of care than whites. PMID- 11886321 TI - Primary human herpesvirus 8 infection in immunocompetent children. AB - CONTEXT: Human herpesvirus 8 (HHV-8) infection causes Kaposi sarcoma and lymphoproliferative disorders in immunosuppressed adults. Its manifestations in immunocompetent hosts are unknown. OBJECTIVES: To determine whether HHV-8 primary infection is symptomatic in immunocompetent children and to identify the epidemiological and virological correlates of HHV-8 infection. DESIGN AND SETTING: Prospective cohort study conducted in the pediatric emergency department of a hospital in Alexandria, Egypt, between December 1, 1999, and April 30, 2000. PATIENTS: Eighty-six children aged 1 to 4 years who were evaluated for a febrile syndrome of undetermined origin. MAIN OUTCOME MEASURES: Serological assay and polymerase chain reaction of blood and saliva samples for HHV-8. Information on potential risk factors for HHV-8 infection was also collected. RESULTS: Thirty six children (41.9%) were seropositive; HHV-8 DNA sequences were detected in 14 (38.9%) of these 36 children (detected in saliva in 11 of 14). Significant associations were found between HHV-8 infection and close contact with at least 2 other children in the community (36 of 63 vs 6 of 23 for <2 children; adjusted odds ratio [OR], 3.50; 95% confidence interval [CI], 1.11-12.22) and admission to the emergency department in December or January (28 of 47 vs 14 of 39 for February-April; adjusted OR, 3.15; 95% CI, 1.23-8.58). Six children had suspected primary HHV-8 infection; all but 1 had a febrile cutaneous craniocaudal maculopapular rash, which was more common among these children (5 of 6 vs 10 of 75; P<.001). For 3 of these 6 children, a second blood sample was obtained after the convalescence phase, and all 3 seroconverted for HHV-8. CONCLUSIONS: Primary infection with HHV-8 may be associated with a febrile maculopapular skin rash among immunocompetent children. The finding of HHV-8 DNA sequences in saliva supports the hypothesis that transmission through saliva is the main mode of transmission in the pediatric age group. PMID- 11886322 TI - Corticosteroid therapy in pulmonary sarcoidosis: a systematic review. AB - CONTEXT: Corticosteroids are used in pulmonary sarcoidosis to reduce symptoms and minimize long-term damage. Spontaneous recovery is a common feature. Both the decision to initiate therapy and the treatment response may be influenced by disease severity, so trials need to use a randomized controlled design. OBJECTIVE: To assess the effect of oral and inhaled corticosteroids on chest radiograph results, symptoms, pulmonary function, and long-term outcome in pulmonary sarcoidosis. DATA SOURCES: MEDLINE, EMBASE, CINAHL, and the Cochrane Controlled Trials Register were searched all years through December 2001. Bibliographies of review articles and retrieved articles were searched, and pharmaceutical companies and authors of identified trials were contacted for other studies. There was no language restriction. STUDY SELECTION: Trials were randomized and included a control group. Participants were adults with histologic evidence of pulmonary sarcoidosis. Treatments included the use of oral and inhaled corticosteroids for at least 8 weeks. The search identified 150 studies; 9 met the inclusion criteria, but only 8 provided usable data. DATA EXTRACTION: Two reviewers assessed trial quality using the Jadad score, which evaluates the quality of randomization, blinding, and reasons for withdrawal. Data were extracted and sent to primary authors for verification. DATA SYNTHESIS: In patients with stage 2 and 3 disease, oral corticosteroids improved findings on the chest radiograph after 6 to 24 months (Peto odds ratio, 2.54; 95% confidence interval [CI], 1.69-3.81; P<.001). Forced vital capacity improved with oral corticosteroids (weighted mean difference [WMD], 4.2% predicted; 95% CI, 0.4% 7.9% predicted) and diffusing capacity also improved (WMD, 5.7% predicted; 95% CI, 1.0%-10.5% predicted). In 2 small studies of inhaled corticosteroids, there was no effect on chest radiograph and inconsistent effects on lung function in one and only a small improvement in symptoms in the other. There were no data following corticosteroid withdrawal to assess any disease-modifying effect. CONCLUSIONS: Oral corticosteroids improved results on the chest radiograph following 6 to 24 months of treatment and produced a small improvement in vital capacity and diffusing capacity. Trials of inhaled corticosteroids were small and results too inconsistent to make firm conclusions concerning their efficacy. There are no data to suggest that corticosteroid therapy alters long-term disease progression. PMID- 11886323 TI - Hypertrophic cardiomyopathy: a systematic review. AB - CONTEXT: Throughout the past 40 years, a vast and sometimes contradictory literature has accumulated regarding hypertrophic cardiomyopathy (HCM), a genetic cardiac disease caused by a variety of mutations in genes encoding sarcomeric proteins and characterized by a broad and expanding clinical spectrum. OBJECTIVES: To clarify and summarize the relevant clinical issues and to profile rapidly evolving concepts regarding HCM. DATA SOURCES: Systematic analysis of the relevant HCM literature, accessed through MEDLINE (1966-2000), bibliographies, and interactions with investigators. STUDY SELECTION AND DATA EXTRACTION: Diverse information was assimilated into a rigorous and objective contemporary description of HCM, affording greatest weight to prospective, controlled, and evidence-based studies. DATA SYNTHESIS: Hypertrophic cardiomyopathy is a relatively common genetic cardiac disease (1:500 in the general population) that is heterogeneous with respect to disease-causing mutations, presentation, prognosis, and treatment strategies. Visibility attached to HCM relates largely to its recognition as the most common cause of sudden death in the young (including competitive athletes). Clinical diagnosis is by 2-dimensional echocardiographic identification of otherwise unexplained left ventricular wall thickening in the presence of a nondilated cavity. Overall, HCM confers an annual mortality rate of about 1% and in most patients is compatible with little or no disability and normal life expectancy. Subsets with higher mortality or morbidity are linked to the complications of sudden death, progressive heart failure, and atrial fibrillation with embolic stroke. Treatment strategies depend on appropriate patient selection, including drug treatment for exertional dyspnea (beta-blockers, verapamil, disopyramide) and the septal myotomy-myectomy operation, which is the standard of care for severe refractory symptoms associated with marked outflow obstruction; alcohol septal ablation and pacing are alternatives to surgery for selected patients. High-risk patients may be treated effectively for sudden death prevention with the implantable cardioverter defibrillator. CONCLUSIONS: Substantial understanding has evolved regarding the epidemiology and clinical course of HCM, as well as novel treatment strategies that may alter its natural history. An appreciation that HCM, although an important cause of death and disability at all ages, does not invariably convey ominous prognosis and is compatible with normal longevity should dictate a large measure of reassurance for many patients. PMID- 11886324 TI - From clinical trials to clinical practice: bridging the GAP. PMID- 11886325 TI - Public profiling of clinical performance. PMID- 11886326 TI - Aging in the 21st century: a call for papers. PMID- 11886333 TI - Harnessing the energy of fusion: Presidential Address, American Head and Neck Society. PMID- 11886334 TI - The dilemma of treating hypopharyngeal carcinoma: more or less: Hayes Martin Lecture. AB - The optimal therapy for hypopharyngeal carcinoma depends on its staging. For early-stage disease, radiotherapy and surgery achieve similar results. Radical surgery followed by radiotherapy is applicable in the management of patients with advanced-stage disease. Chemoradiation aiming to preserve the larynx can only be performed for selected patients and in well-equipped institutions. Thorough understanding of pathological behavior of hypopharyngeal carcinoma, its submucosal tumor extension, and its high propensity to metastasize to cervical lymph nodes allows head and neck surgeons to choose optimal surgical treatment. Lymph node status determines the type of neck dissection required while location and size of the primary tumor determine the extent of resection and choice of reconstruction procedure. Adequate tumor extirpation with less extensive and invasive procedures preserving unaffected normal tissue contribute to more tumor control and less morbidity. PMID- 11886335 TI - James Barrett Brown (1899-1971), head and neck surgeon of a half century ago. AB - This article summarizes the life and work of James Barrett Brown, MD (1899-1971), a plastic surgeon from St Louis, Mo, whose many contributions to the knowledge of surgery include his pioneering use of large split-thickness skin grafts to resurface defects. Along with a coauthor, he published an excellent book on radical neck dissection in 1954 (Neck Dissections, published by Charles C. Thomas, Springfield, Ill). Brown was a leading figure in the organization of high quality plastic surgical care to injured soldiers in World War II. His training program in plastic surgery at Barnes Hospital in St Louis provided education to many leaders in the field. He received a number of honors for his many accomplishments. PMID- 11886336 TI - Hurthle cell tumors: using molecular techniques to define a novel classification system. AB - BACKGROUND: Since ret/PTC gene rearrangements are specific to papillary thyroid carcinoma (PTC), the diagnosis of Hurthle cell PTC (HCPTC) has recently been expanded to include a subset of Hurthle cell tumors (HCTs) that may lack both papillary architecture and/or classic nuclear features but that harbor a ret/PTC gene rearrangement. We hypothesize that such HCPTCs behave in a fashion analogous to other papillary carcinomas, while Hurthle cell carcinomas (HCCs) behave similarly to follicular carcinomas. EDUCATIONAL OBJECTIVES: At the conclusion of this article, participants should be able to discuss HCTs and to identify HCPTCS using molecular techniques. METHODS: A retrospective chart review was carried out on 56 patients with HCTs. All pathological specimens were analyzed for ret/PTC gene rearrangements. Hurthle cell adenoma (HCA) was defined as an HCT that did not exhibit capsular and/or vascular invasion and that lacked a ret/PTC gene rearrangement when evaluated by immunohistochemical and reverse transcription polymerase chain reaction analysis. An HCC was defined as an HCT with capsular and/or vascular invasion that lacked a ret/PTC gene rearrangement, and an HCPTC was defined as any HCT that harbored a ret/PTC gene rearrangement. RESULTS: The subclassification of the 56 HCTs was as follows: 21 HCAs, 15 HCCs, and 20 HCPTCs. No patients with HCA or HCC were ret/PTC positive. Five of the 6 patients with definite lymph node metastasis were in the HCPTC group, demonstrating that molecular analysis helps to explain biological behavior. CONCLUSIONS: Hurthle cell neoplasms can now be classified using histopathological as well as molecular criteria. It appears that the new subclassification of malignant HCTs into follicular (HCC) and papillary (HCPTC) variants identifies 2 distinct biological groups. PMID- 11886337 TI - A prospective study of intraoperative lymphatic mapping for head and neck cutaneous melanoma. AB - BACKGROUND: Intraoperative lymphatic mapping and sentinel lymph node biopsy have been used successfully to stage regional lymphatics for trunk and extremity melanomas. However, the accuracy and applicability of these techniques in the head and neck have not been determined conclusively. OBJECTIVE: To report the results of a prospective trial of intraoperative lymphatic mapping and sentinel lymph node identification in patients with head and neck cutaneous melanoma. METHODS: Using technetium Tc 99m--labeled sulfur colloid and isosulfan blue, intraoperative lymphatic mapping and sentinel lymph node identification were performed in 43 patients with melanomas of intermediate thickness. After the sentinel lymph nodes were identified in situ, an elective dissection of levels I through V or II through V was performed, based on the location of the primary tumor. The parotid, postauricular, and suboccipital lymphatics were dissected as clinically indicated. The sentinel lymph nodes were isolated ex vivo and evaluated pathologically by serial sectioning, and the accuracy of the lymphatic mapping was determined. RESULTS: Intraoperative lymphatic mapping identified 155 sentinel lymph nodes in 94 nodal basins, with a mean of 3.6 sentinel nodes and 2.2 basins per patient. Sentinel nodes were located in the parotid gland in 19 patients (44%), necessitating superficial parotidectomies, and they were distributed throughout nonadjacent nodal basins in 18 patients (42%). Nine patients (21%) had metastatic disease in 1 or more sentinel nodes, 3 of whom had metastatic disease in a nonsentinel node. No patient who had negative sentinel nodes had a positive nonsentinel node (false-negative incidence, 0). CONCLUSIONS: Although intraoperative lymphatic mapping accurately identifies sentinel lymph nodes for head and neck cutaneous melanomas, the multiplicity of these nodes, their widespread distribution, and their frequent location within the parotid gland may preclude sentinel lymph node biopsy in many patients. Therefore, we advocate selective lymphadenectomy of sentinel nodal basins, allowing histological staging of the regional lymphatics with limited morbidity. However, further study is necessary to define the true role of sentinel lymph node identification for head and neck cutaneous melanoma. PMID- 11886338 TI - Pulmonary atelectasis after reconstruction with a rectus abdominis free tissue transfer. AB - BACKGROUND: Atelectasis is one of the most common postoperative complications encountered in head and neck surgery. Risk factors include preexisting pulmonary disease, the procedure performed, and the length of anesthetic. Regional flaps used to reconstruct defects in the head and neck predispose to radiographic atelectasis. The rectus abdominis myocutaneous flap is usually transferred as a free tissue transfer. Harvesting the flap results in abdominal wall pain and postoperative splinting that may contribute to an increased development of atelectasis. To our knowledge, this issue has not been previously examined. DESIGN: Retrospective review. RESULTS: Fifty-three patients underwent rectus abdominis myocutaneous free flap reconstruction following major ablative procedures for head and neck cancer. The flap size ranged from 5 x 7 to 25 x 27 cm. Most flaps were 8 x 15 cm. The cutaneous area transferred ranged from 35 to 600 cm(2) (mean, 120 cm(2)). These patients were compared with a group of 53 patients who were matched for age, sex, length of the procedure, and stage of disease. Postoperative atelectasis was radiographically detected in 37 (70%) of the patients who underwent rectus abdominis myocutaneous free flap reconstruction vs 41 (77%) of the controls. Major atelectasis was not encountered in any patient in either group. Patients with a larger cutaneous paddle (>120 cm(2)) had a higher atelectasis score than patients with smaller cutaneous paddles (< or =120 cm(2)) (P =.02). CONCLUSIONS: The incidence of radiographic postoperative atelectasis in patients undergoing rectus abdominis myocutaneous free tissue transfer is high. The degree of atelectasis is small, and the clinical correlation and relevance are minimal. PMID- 11886340 TI - Clinical care pathway for head and neck cancer: a valuable tool for decreasing resource utilization. AB - OBJECTIVE: To evaluate the durability over time of the reduction of resource utilization after implementing a clinical care pathway (CCP) for head and neck cancer surgery. DESIGN: Cohort study. SETTING: A tertiary care academic medical center. PATIENTS: We studied control subjects from 1995 (pre-CCP) (n = 87), a cohort from July 1, 1996, through July 31, 1997 (the first year after CCP implementation) (n = 43), and a cohort from 1999 (n = 82) after major resection and tracheostomy for upper aerodigestive tract cancer. INTERVENTIONS: Starting July 1, 1996, all patients undergoing major resection for head and neck cancer were treated using a CCP, which delineates daily interventions and goals. MAIN OUTCOME MEASURES: Length of stay (LOS), readmission and complication rates, and hospital charges. RESULTS: Median total LOS and LOS exclusive of the intensive care unit decreased in the first year and remained stable at 3 years (from 13.0 to 8.0 days and from 10.5 to 6.4 days, respectively). The intensive care unit LOS decreased across 3 years from 2.2 to 1.1 days (P=.001). Median total charges declined from 105,410 US dollars pre-CCP to 65,919 US dollars at 3 years. Incidence of postoperative pneumonia decreased from 12% to 1% (P=.02), and readmission rate decreased from 18% to 11% (P=.37) across 3 years. CONCLUSIONS: The CCP for head and neck cancer maintained the improvement in LOS and charges seen in the first year of implementation and continues to decrease resource utilization while enhancing quality of care. PMID- 11886339 TI - Underexpression of p27/Kip in thyroid papillary microcarcinomas with gross metastatic disease. AB - OBJECTIVE: Papillary microcarcinomas (PMCs) of the thyroid (measuring less than 1 cm in maximum dimension) are extremely common incidental histologic findings, and most of these tumors are not considered clinically significant. However, rare PMCs behave aggressively and metastasize early, giving rise to clinically significant metastatic disease. We hypothesized that p27 and MIB-1/Ki-67 immunoreactivity would allow us to identify this small subgroup of PMCs that have the potential to behave aggressively. METHODS: We reviewed the histopathology reports of 2000 patients who underwent thyroid surgery at our institution between 1995 and 1999 and identified 22 patients who presented with gross regional metastases from a primary PMC. The primary and metastatic tumors were stained for ret, p53, p27, and MIB-1 using the avidin-biotin-peroxidase complex technique. A control group of 33 nonmetastasizing PMCs was also analyzed. RESULTS: Immunoreactivity for ret, p53, and MIB-1 showed no difference between metastasizing and nonmetastasizing PMCs. In most tumors, ret was present, while p53 immunoreactivity was absent in all tumors. MIB-1 staining was present in a small number of cells in both groups of tumors. Immunoreactivity for p27 was quantitated by the intensity of expression as well as the distribution of positive cells within each tumor. All tumors showed lower p27 expression than normal thyroid tissue. However, metastasizing PMCs demonstrated a significantly lower expression of p27 than nonmetastasizing PMCs (P<.001). CONCLUSION: Our results suggest that p27 immunohistochemical analysis may be a valuable diagnostic tool in predicting aggressive potential in PMCs. PMID- 11886341 TI - The value of frozen section examinations in determining the extent of thyroid surgery in patients with indeterminate fine-needle aspiration cytology. AB - OBJECTIVES: To determine the usefulness of intraoperative frozen section (FS) examinations in establishing the diagnosis of thyroid cancer in patients undergoing thyroidectomy for nodules with indeterminate cytological features and to determine the cost-effectiveness of FS examinations in this situation. DESIGN: Retrospective medical record review. The results of fine-needle aspiration biopsies (FNABs), FS examinations, and final pathologic examinations are compared. A cost-effectiveness analysis of routine FS examinations compared with the cost of additional surgical procedures is performed. SETTING: A private surgical practice in a medical school-affiliated teaching hospital. PATIENTS: The records of all 480 patients undergoing thyroidectomy between January 1, 1998, and September 30, 2000, were reviewed. All 199 patients with a dominant thyroid nodule and FNAB results either highly suggestive of papillary cancer or indeterminate were studied. RESULTS: Of the patients with FNAB results highly suggestive of papillary cancer, 95% had cancer according to the final pathologic examination results. The diagnosis of cancer was made by FS examination results in 67% of these patients. Of the remaining 178 patients whose FNAB result was indeterminate, 64 (36%) had thyroid cancer. Malignancy was diagnosed by FS examination results in 30 (47%) of these patients. If FS examinations had not been performed, these 30 patients would have required a second operation to complete a total thyroidectomy. The cost savings of routine FS examinations in patients with indeterminate FNAB results is 1298 US dollars per patient. CONCLUSIONS: The routine performance of FS examinations in patients with thyroid nodules with indeterminate cytological features is a cost-effective way of avoiding a second surgical procedure if a total thyroidectomy is indicated. In patients with FNAB results highly suggestive of papillary cancer, FS examinations are not useful. In these patients, the definitive operation can be based on the results of the FNAB. PMID- 11886342 TI - Head and neck cancer incidence trends in young Americans, 1973-1997, with a special analysis for tongue cancer. AB - OBJECTIVE: To examine the temporal changes in head and neck cancer in young adults in the United States. METHODS: Using the cancer surveillance database from the National Cancer Institute Surveillance, Epidemiology, and End Results (SEER) Program, we calculated age-adjusted incidence rates for head and neck cancers. Using the joinpoint regression model, we described tongue cancer incidence trends and established the statistical significance of temporal changes. We also compared changes in 5-year survival rates for tongue cancer. RESULTS: From 1973 to 1997, there were 63 409 patients with head and neck cancer in the 9 SEER registries. Of these, 3339 patients were younger than 40 years. The incidence of head and neck cancer remained stable in groups older than 40 years comparing the 1973-1984 and 1985-1997 data. In contrast, tongue cancer in adults younger than 40 years increased approximately 60% during the same period. We detected a significant increase until 1985, the estimated annual percentage change being 6.7% (95% confidence interval, 2.7%-10.8%; P<.001). After 1985, incidence rates stopped rising but remained steadily high. The change in tongue cancer incidence rates for young adults was related to birth cohorts between 1938 and 1948. The absolute increase in 5-year survival for tongue cancer ranged from 11.7% (<40 years old) to 6.6% (40-64 years old) between 1973-1984 and 1985-1997, with the most significant improvement occurring in young Americans with regional or distant disease (27% and 21%, respectively). CONCLUSIONS: A sharp increasing trend in tongue cancer in young Americans may be attributed to persons born after 1938. The reason for the increase is uncertain. Improved survival rates in young patients suggest the emergence of a distinct disease process that is apparent in white but not black populations. PMID- 11886343 TI - Persistent parathyroid hormone elevation following curative parathyroidectomy for primary hyperparathyroidism. AB - BACKGROUND: Persistent elevation of parathyroid hormone (PTH) levels following parathyroidectomy may indicate residual abnormal parathyroid tissue. OBJECTIVE: To determine the clinical significance and risk factors for persistent PTH elevation following curative parathyroidectomy. METHODS: A prospective study of consecutive patients with primary hyperparathyroidism who had resolution of hypercalcemia following parathyroidectomy. Patients with low or normal serum calcium and increased PTH levels postoperatively were identified, and serial calcium and PTH levels and clinical course were monitored. A multivariate analysis was performed to identify features associated with an elevated postoperative PTH level. RESULTS: Of 85 patients with resolution of hypercalcemia following parathyroidectomy, postoperative PTH levels were elevated in 23 (27%) (mean, 99 pg/mL; range, 70-194 pg/mL) and normal in 62 (mean, 30 pg/mL; range, 3 65 pg/mL) (P<.001). No significant differences in preoperative or postoperative calcium or preoperative PTH levels were found between groups. Among patients with persistent PTH elevation, 18 had adenoma and 5 had multiglandular disease, compared with 52 with adenoma and 10 with multiglandular disease in patients with normal postoperative PTH levels (P>.05). Multivariate analysis demonstrated that black race and musculoskeletal symptoms were associated with an elevated postoperative PTH level (P =.01. After an average 16-month follow-up, PTH levels normalized in 13 patients, decreased in 5, and were unchanged in 2. Three patients were lost to follow-up. CONCLUSIONS: Persistent PTH elevation occurs in 27% of patients following curative parathyroidectomy and is usually a transient phenomenon more common in patients with musculoskeletal symptoms and of the black race. It is not a manifestation of persistent disease but is most likely a secondary response to bone remineralization. PMID- 11886344 TI - Nasopharyngectomy for recurrent nasopharyngeal cancer: a 2- to 17-year follow-up. AB - OBJECTIVE: To review the 2- to 17-year outcome of nasopharyngectomy following local recurrence of nasopharyngeal carcinoma. DESIGN: Retrospective review. SETTING: University medical center. PATIENTS: Thirty-seven patients with biopsy proven recurrent nasopharyngeal cancer followed up for a minimum of 2 years after transpalatal, transmaxillary, and/or transcervical resection with and without neck dissection. OUTCOME: Clinical examination, magnetic resonance imaging, chest x-ray examination, and liver function tests to determine re-recurrence; unlimited follow-up. RESULTS: With a mean follow-up of 5.4 years, the crude, 5-year, overall, free-of-disease survival rate was 52%, local control at 5 years was 67%, and the 5-year actuarial survival rate was 60%. Survival by recurrent T stage (rT) was as follows: rT1, 73%; rT2, 40%; rT3, 14%; and rT4, 0%. Complications occurred in 54% and included 1 death from carotid artery injury and 1 patient with permanent pharyngeal plexus paralysis with resultant dysphagia. The remaining patients had transitory complications that spontaneously resolved, required further surgery (closure of palate fistula, debridement, and reapplication of skin graft), or required further medical therapy. CONCLUSIONS: The results of this study are better than most published reports of additional irradiation for rT1 and rT2 lesions. More recent radiation studies that use radiosurgery or implants suggest promising early results. A randomized prospective study comparing surgery with additional irradiation for recurrent disease at the primary site is warranted. PMID- 11886345 TI - Sentinel lymph node biopsy for cutaneous head and neck melanomas. AB - OBJECTIVE: To report the results of sentinel lymph node biopsy (SLNB) for cutaneous head and neck melanomas (CMHNs). DESIGN: Consecutive series followed for a median of 20 months. SETTING: Tertiary cancer care center. PATIENTS: Fifty six individuals with clinically node-negative CMHN, median Breslow thickness, 2.6 mm (range, 0.2-20.0 mm). INTERVENTIONS: Preoperative technetium 99m sulfur colloid lymphoscintigraphy (PLSG) followed within 4 hours by intraoperative handheld gamma probe localization (IHGP). Intraoperative injection of 1% isosulfan blue dye (IBD) was used in 48 patients. Immediate completion nodal dissection was performed for metastatic SLNs on intraoperative frozen section analysis and monitoring for negative SLNs. MAIN OUTCOME MEASURES: Rate of SLN identification, SLN and non-SLN positivity, same-basin recurrence, and disease specific and recurrence-free survival. RESULTS: Combination of IHGP and IBD improved SLN identification to 96% from 93% for IHGP and 73% for IBD alone. Four patients had a positive SLN on frozen section analysis. A negative SLNB correctly predicted regional nodal control in 47 of 48 patients but missed 1 of 5 patients who had regional lymphatic disease. All 4 patients who failed SLNB remain alive and free of recurrent disease. Two-year Kaplan-Meier disease-specific and relapse free survival was 91% and 88%, respectively. Two-year disease-specific survival was 93% for SLN-negative patients and 50% for SLN-positive patients (P=.20). CONCLUSIONS: Combining PLSG with IHGP and IBD improves the success rate of SLNB. Although SLNB is a reliable indicator of the status of the draining lymphatic basins in CMHN, patients with negative SLNs must be observed for longer periods to understand the true implications of the procedure. PMID- 11886346 TI - Recurrence rates after selective neck dissection in the N0 irradiated neck. AB - OBJECTIVES: To define patterns of subclinical metastases in irradiated N0 necks with recurrent or persistent primary site disease and to determine the regional control rate when selective neck dissection (SND) is used in this setting. PATIENTS AND INTERVENTION: Individuals included were previously treated for head and neck squamous cell carcinoma with primary radiation therapy or chemoradiotherapy. All had recurrent or persistent disease at the primary site, with no clinical or radiographic evidence of nodal disease. The patients underwent surgical treatment of the primary site along with site-specific SND and were required to undergo at least 1 year of follow-up. Subsequent recurrence at the primary site disqualified the patient from further evaluation. MAIN OUTCOME MEASURE: Regional tumor control. RESULTS: Forty-three patients meeting the inclusion criteria underwent 59 SNDs (levels dissected: I-IV [n = 22], II-IV [n = 34], and I-III [n = 3]). Sixteen specimens were positive for nodal disease. The charts of 26 patients, who underwent a total of 35 SNDs, were available for review after 1 year (none of the patients involved died of disease in the neck). There were no neck recurrences (mean follow-up, 25 months; median, 21 months). All patients with more than 2 occult nodal metastases experienced primary site recurrence or distant metastases. CONCLUSIONS: In this small cohort, SND in previously irradiated patients with recurrent primary disease but clinically negative necks has resulted in excellent tumor control in the neck. The usual patterns of nodal spread do not appear to be significantly altered with primary site recurrence after radiation therapy. The presence of more than 2 positive nodes in the neck specimen correlates with poor prognosis. PMID- 11886347 TI - Superior laryngeal nerve identification and preservation in thyroidectomy. AB - BACKGROUND: Injury to the external branch of the superior laryngeal nerve (EBSLN) can result in detrimental voice changes, the severity of which varies according to the voice demands of the patient. Variations in its anatomic patterns and in the rates of identification reported in the literature have discouraged thyroid surgeons from routine exploration and identification of this nerve. Inconsistent with the surgical principle of preservation of critical structures through identification, modern-day thyroidectomy surgeons still avoid the EBSLN rather than identifying and preserving it. OBJECTIVES: To describe the anatomic variations of the EBSLN, particularly at the junction of the inferior constrictor and cricothyroid muscles; to propose a systematic approach to identification and preservation of this nerve; and to define the identification rate of this nerve during thyroidectomy. MATERIALS AND METHODS: A retrospective review of thyroid lobectomies and total thyroidectomies performed between 1978 and 1997 was carried out. A total of 884 patients were included, with 1057 EBSLNs explored. Intraoperative findings of identification of the EBSLN were recorded and compared on an annual basis for both benign and malignant disease. Overall results were also compared with those found in previous series identified through a 50-year literature review. RESULTS: The 3 anatomic variations of the distal aspect of the EBSLN as it enters the cricothyroid were encountered and are described. The total identification rate over the 20-year period was 900 (85.1%) of 1057 nerves. Operations performed for benign disease were associated with higher identification rates (599 [86.1%] of 696) as opposed to those performed for malignant disease (301 [83.4%] of 361). Operations performed in recent years have a higher identification rate (over 90%). CONCLUSIONS: Understanding the 3 anatomic variations of the distal portion of the EBSLN and its relation to the inferior constrictor muscle allows for high rates of identification of this nerve. The EBSLN should be explored during thyroid surgery and identification is possible in most cases. Preservation of the EBSLN maintains optimal function of the larynx. PMID- 11886348 TI - Paraglottic space in supracricoid laryngectomy. AB - BACKGROUND: Paraglottic space (PGS) is a connective tissue compartment of the larynx and is important in the extension of laryngeal cancer. It communicates with the preepiglottic space superiorly and with the extralaryngeal region inferiorly through the gap within the cricothyroid membrane. Transglottic cancer of the larynx, which spreads within PGS, is characterized by a high incidence of laryngeal skeleton invasion and of cervical metastasis. Determining the correct stage of transglottic cancer of the larynx is difficult, leading to therapeutic failure of partial laryngectomy in some cases. OBJECTIVE: To clinically confirm a pathologically complete resection of PGS from the piriform sinus mucosa by supracricoid partial laryngectomy in laryngeal cancers involving PGS. MATERIALS AND METHODS: Eight patients with transglottic cancer whose cancer was confirmed clinically and pathologically at stages T2b or higher underwent supracricoid partial laryngectomy. During supracricoid partial laryngectomy, we performed a sharp dissection of PGS from the piriform sinus mucosa to obtain a complete resection margin while preserving the piriform sinus mucosa. Microscopic evaluation of the specimens was made for the invasion of PGS and the safe margin distance from the piriform sinus mucosa. RESULTS: Pathological cancer invasion of PGS was confirmed in 7 of 8 patients and a sufficient pathological margin from tumor invasion to the piriform sinus mucosa was obtained. The average safety margin was 10.3 mm. CONCLUSION: Supracricoid partial laryngectomy could be considered a safe surgical modality for cancers not extending to PGS. PMID- 11886349 TI - Effect of blood transfusion in an experimental sarcoma model. AB - OBJECTIVE: To study the effect of allogeneic, syngeneic, and autologous blood transfusion on the growth rate of the KHT tumor in a C3H murine model. DESIGN: Prospective, randomized, and controlled animal study. SUBJECTS: Sixty-one C3H female mice. INTERVENTIONS: The C3H female mice were implanted with 2 x 10(5) cells of KHT, a murine sarcoma. Ten days later, 0.3 mL of blood was removed from a retro-orbital site to simulate surgical blood loss. This blood loss was replaced by blood transfusion through a tail vein with the use of allogeneic (major histocompatibility complex incompatible), syngeneic (major histocompatibility complex compatible), or autologous blood. Tumor growth was measured daily for 14 days. The tumor growth curve for each of the animals was constructed and the mean slope of growth calculated for each group. RESULTS: There were statistically significant differences in tumor growth rate (P =.001) when the allogeneic group (mean slope = 0.232, n = 14), the syngeneic group (mean slope = 0.190, n = 17), and the autologous group (mean slope = 0.202, n = 14) were compared. A t test confirmed that there was no significant difference in the tumor growth rate between the groups transfused with syngeneic and autologous blood (P =.26). However, the rate of tumor growth in the allogeneic group was found to be significantly higher when independently compared with the syngeneic group (P<.001) and the autologous group (P =.02). CONCLUSIONS: In this experimental model of a solid murine sarcoma, allogeneic blood transfusion was associated with an increased rate of tumor growth compared with syngeneic and autologous blood transfusion, likely reflecting immunomodulatory effects incurred by the introduction of major histocompatibility complex-incompatible antigens. PMID- 11886350 TI - Predictive factors for diagnosis of advanced-stage squamous cell carcinoma of the head and neck. AB - OBJECTIVE: To identify the predictive factors (with emphasis on diagnostic delay) associated with the diagnosis of an advanced-clinical stage head and neck cancer. DESIGN: Cross-sectional study of patients with head and neck cancer originally recruited for a case-control study. SETTING: Three referral oncological centers in metropolitan areas in southern Brazil: Sao Paulo, Curitiba, and Goiania. PATIENTS: The study population comprised 679 patients recently diagnosed as having a previously untreated head and neck squamous cell carcinoma. MAIN OUTCOME MEASURE: Diagnosis of advanced disease (clinical stage III-IV) head and neck cancer. RESULTS: Patients with laryngeal and hypopharyngeal cancers were more likely to be diagnosed as having advanced disease than those with lip, oral, and oropharyngeal cancers (88.0% vs 74.6%) (P<.001). Patient delay was inversely associated with clinical stage at diagnosis in patients with the same cancers, while professional delay was directly associated with a higher risk of advanced clinical stage at diagnosis (P =.001 and P =.006, respectively). In the analysis of laryngeal and hypopharyngeal cancer, both patient and professional delays were associated with advanced disease, with patient delay being a stronger predictive factor than professional delay. CONCLUSIONS: Clinical stage at diagnosis was associated with sociodemographic characteristics, patient delay, and professional delay. Our results indicate that continued educational programs for the population and health care professionals regarding the identification of early symptoms of head and neck cancers are warranted. PMID- 11886351 TI - The thoracoacromial/cephalic vascular system for microvascular anastomoses in the vessel-depleted neck. AB - OBJECTIVE: To review our experience with use of the thoracoacromial/cephalic (TAC) system in the free flap reconstruction of complicated head and neck defects. DESIGN: Case series. SETTING: Tertiary care referral center. POPULATION: A consecutive sample of 11 patients requiring free flap reconstruction of head and neck defects using the TAC system for microvascular anastomoses was identified by medical chart review. INTERVENTION: Free flap reconstruction of complicated defects of the head and neck using the TAC vascular system for microvascular anastomoses. MAIN OUTCOME MEASURES: Free flap survival and microvascular thrombosis. RESULTS: Of 11 patients using TAC anastomoses, all had complete survival of free flaps. No complications related to anastomotic failure were identified. CONCLUSIONS: The TAC system provides a reliable source of undisturbed vessels when cervical vessels are unusable or absent. PMID- 11886352 TI - Changing patterns of failure of head and neck cancer. AB - BACKGROUND: With the increased use of neoadjuvant therapy for advanced stage squamous cell carcinoma of the head and neck, we have observed an apparent change in the pattern of failure from predominantly locoregional sites to distant metastases. We reviewed the patterns of failure in cancers of the oral cavity, oropharynx, and larynx at our institution during the last decade. OBJECTIVE: To determine whether there has been a significant change in the patterns of recurrence from the historical locoregional failure to distant sites, and whether this change is associated with the increased use of multimodality therapy. METHODS: We reviewed cancer registry data on patients with squamous cell carcinoma of the head and neck diagnosed between January 1, 1988, and December 31, 1999. Sites included the oral cavity and oropharynx (including the tongue, floor of mouth, retromolar trigone, gingiva, tonsil, and lip) and larynx. RESULTS: Among 432 patients with squamous cell carcinoma of the head and neck, 280 (65%) had oral cavity and oropharyngeal cancers, and 152 (35%) had laryngeal cancers. Overall, 19% developed locoregional recurrence, and 8% developed distant failure. Although locoregional failure for oral cavity and oropharyngeal squamous cell carcinoma decreased from 26% to 16% from 1988-1993 to 1994-1999, distant failure increased significantly from 3% to 8%. During these periods, multimodality therapy was used in 62% of oral cavity and oropharyngeal cancers, and this rate remained essentially unchanged. For laryngeal cancer, locoregional and distant failure remained stable at 18% and 9%, respectively. In these laryngeal cancers, the use of multimodality therapy decreased from 60% to 46%, but this difference was not statistically significant (P =.43). CONCLUSIONS: Although locoregional control in oral cavity and oropharyngeal cancers has improved significantly with the use of multimodality therapy, the incidence of distant failure has doubled. In laryngeal squamous cell carcinoma, the patterns of failure have not changed significantly. PMID- 11886353 TI - Microvascular reconstruction after previous neck dissection. AB - BACKGROUND: Microvascular reconstruction of defects in the head and neck is more challenging in patients who have undergone a previous neck dissection, owing to prior resection of potential cervical recipient blood vessels used for free flap perfusion. OBJECTIVE: To evaluate the reliability and safety of free flap reconstruction in patients with previous neck dissection. PATIENTS AND METHODS: Sixty free flaps were performed in 59 patients with a medical history of neck dissection for head and neck cancer. This included patients undergoing salvage surgery for recurrent cancer as well as patients undergoing secondary reconstruction of cancer surgery-related defects. Flap selection included 25 radial forearm flaps, 20 fibula flaps, 7 rectus abdominis flaps, 7 subscapular system flaps, and 1 iliac crest flap. RESULTS: Recipient vessels were used in the field of previous neck dissection in approximately half the patients with previous selective neck dissection, while contralateral recipient vessels were always used in patients with a history of modified radical or radical neck dissection. Vein grafts were not necessary in any cases. One arterial anastomosis that was created under excessive tension required urgent reoperation and revision, but there were no cases of free flap failure. CONCLUSIONS: Free flap reconstruction of the head and neck is highly successful in patients with a history of neck dissection, despite a relative paucity of potential cervical recipient blood vessels. Heavy reliance on free flaps with long vascular pedicles obviated the need to perform vein grafts in the present series, probably contributing to the absence of free flap failure. Previous neck dissection should not be considered a contraindication to microvascular reconstruction of the head and neck. PMID- 11886354 TI - Lhermitte-Duclos disease (dysplastic cerebellar gangliocytoma): a malformation, hamartoma or neoplasm? AB - OBJECTIVES: Dysplastic gangliocytoma (Lhermitte-Duclos disease) is a rare disorder, characterized by a slowly progressive unilateral tumour mass of the cerebellar cortex. The fundamental nature of this apparently benign entity and in particular its pathogenesis remain unknown. The debate, whether it represents a neoplastic, malformative or hamartomatous lesion, is still in progress. Lhermitte Duclos disease was recently encountered to be part of a multiple hamartoma neoplasia complex (Cowden's syndrome). METHODS: The present account gives a review of the pertinent literature with emphasize on clinical presentation, radiological findings, surgical procedures, histopathological features and pathogenetic considerations of dysplastic cerebellar gangliocytoma. RESULTS: Dysplastic cerebellar gangliocytoma clusters within the third to fourth decades of life. Cranial nerve palsies, unsteadiness of gait, ataxia and sudden neurological deterioration as a result of occlusive hydrocephalus are frequent signs and symptoms. Associations with other congenital malformations, such as megalencephaly, polydactylia, multiple haemangioma and skull abnormalities are common. Magnetic resonance imaging (MRI) is the diagnostic modality of choice and reveals characteristic non-enhancing gyriform patterns with enlargement of cerebellar folia. Surgery is the therapeutic procedure generally performed and complete resection was attempted in the majority of cases. The histopathological findings of Lhermitte-Duclos disease include widening of the molecular layer, which is occupied by abnormal ganglion cells, absence of the Purkinje cell layer and hypertrophy of the granule cell layer. CONCLUSIONS: Dysplastic gangliocytoma of the cerebellum is of benign behaviour and its incidence is extremely rare. The disease should be considered when confronted with a young adult presenting with clinical signs of progressive mass effect in the posterior fossa. The lesion is hypointense on T1- and hyperintense on T2-weighted magnetic resonance images. Recognition of the disease is of particular importance, as the frequent but under reported coexistence with Cowden syndrome, should prompt thorough clinical and apparative investigation to detect or exclude concomitant malignancies. PMID- 11886355 TI - Focal enhancement of motor cortex excitability during motor imagery: a transcranial magnetic stimulation study. AB - OBJECTIVES: In order to learn more about the physiology of the motor cortex during motor imagery, we evaluated the changes in excitability of two different hand muscle representations in the primary motor cortex (M1) of both hemispheres during two imagery conditions. MATERIALS AND METHODS: We applied focal transcranial magnetic stimulation (TMS) over each M1, recording motor evoked potentials (MEPs) from the contralateral abductor pollicis brevis (APB) and first dorsal interosseus (FDI) muscles during rest, imagery of contralateral thumb abduction (C-APB), and imagery of ipsilateral thumb abduction (I-APB). We obtained measures of motor threshold (MT), MEP recruitment curve (MEP-rc) and F waves. RESULTS: Motor imagery compared with rest significantly decreased the MT and increased MEPs amplitude at stimulation intensities clearly above MT in condition C-APB, but not in condition I-APB. These effects were not significantly different between right and left hemisphere. MEPs simultaneously recorded from the FDI, which was not involved in the task, did not show facilitatory effects. There were no significant changes in F wave amplitude during motor imagery compared with rest. CONCLUSIONS: Imagery of unilateral simple movements is associated with increased excitability only of a highly specific representation in the contralateral M1 and does not differ between hemispheres. PMID- 11886356 TI - A transcranial magnetic stimulation study evaluating methylprednisolone treatment in multiple sclerosis. AB - OBJECTIVE: To investigate the efficacy of two different high doses of intravenous methylprednisolone (IVMP) during Multiple Sclerosis (MS) relapses. BACKGROUND: Transcranial Magnetic Stimulation (TMS) is the most sensitive neurophysiological ascertainment to quantify motor disability, to follow the recovery from an MS relapse, and to detect the response to treatment. DESIGN AND METHOD: Twenty-four clinically definite relapsing - remitting MS patients presenting a relapse were randomly assigned to a treatment for 5 days with IVMP 1 or 2 g/day. The response to treatment of each patient was evaluated through Expanded Disability Status Scale (EDSS), Medical Research Council (MRC) score, and TMS by means of motor evoked potential (MEP) parameters. RESULTS: Motor threshold (MT), central motor conduction time (CMCT) and MRC showed a higher improvement with the highest dose of IVMP. Silent period and EDSS improved with both treatments. CONCLUSION: The dose of 2 g/day of IVMP is more effective in MS relapse. PMID- 11886357 TI - Intrathecal IgG synthesis: marker of progression in multiple sclerosis patients. AB - OBJECTIVES: We study the power of IgG synthesis value as a marker of disease activity in multiple sclerosis (MS). MATERIAL AND METHODS: Link index was calculated in 202 MS patients. Time between first, second and third attack and progression index (PI) were compared in patient with normal (NLI) high (HL) or very high Link index (VHLI). RESULTS: Secondary progressive (SP) patients had a higher LI than relapsing-remitting (RR) and primary progressive (PP) courses (1.10 +/- 0.5 for SP vs 0.86 +/- 0.5 for RR and 0.81 +/- 0.5 for PP, P=0.01 and 0.03, respectively). Having a HLI in MS RR and SP patients has no time effect in the development of the second and third attack. PI was higher in patients with VHIL (0.67 +/- 0.7) vs patients with NLI (0.42 +/- 0.4, P=0.008) and with HLI (0.39 +/- 0.3, P=0.001). CONCLUSIONS: This study confirmed that LI is a good marker of subsequent progression of MS. PMID- 11886358 TI - Changes of the MS functional composite and EDSS during and after treatment of relapses with methylprednisolone in patients with multiple sclerosis. AB - OBJECTIVES: The Multiple Sclerosis Functional Composite (MSFC) comprises quantitative functional measures of leg, hand/arm and cognitive function. We examined the responsiveness of the MSFC compared with the Expanded Disability Status Scale (EDSS) during treatment of relapses in patients with multiple sclerosis (MS). PATIENTS AND METHODS: 27 patients received 1000 mg intravenous methylprednisolone (i.v.-MP) for 5 days, followed by oral methylprednisolone for 14 days. The MSFC and the EDSS-score were assessed on day 0, before the first corticosteroid treatment, on day 5, after the last course of i.v. MP, and on day 20 after the treatment was finished. Before the first administration of the MSFC, patients were trained for the paced auditory addition test (PASAT) performing three test trials. In order to analyse practice effects, 10 MS patients without an acute exacerbation were tested three times under the same conditions as the treated group. RESULTS: The median EDSS-score was 2.5 in both groups. On day 5 it remained unchanged in all treated patients, on day 20 a decrease of 0.5 EDSS point occurred in five patients, and in two patients an improvement with a decrease of more than 0.5 point was observed. There was no statistically significant difference between the EDSS-scores on day 0, 5 and 20. The mean MSFC score in the treated group was -0.14 +/- 0.63 on day 0, 0.17 +/- 0.66 on day 5, and 0.42 +/- 0.59 on day 20. On the last study day, 26 patients improved compared with day 0. The differences between the MSFC-scores at the three points of time were statistically significant for the treated group (P < 0.001), but not for the control group. CONCLUSION: During and after treatment of relapses in patients with MS, the MSFC appears to be more sensitive in detecting changes in function than the EDSS. PMID- 11886359 TI - Driving accident frequency increased in patients with multiple sclerosis. AB - OBJECTIVES: To assess the influence of multiple sclerosis (MS) on the ability to drive safely. MATERIAL AND METHODS: A 10-year historical cohort register-study on 197 patients with MS and 545 controls individually matched on age, gender, place of residence, and exposure period. Persons with other neurological diseases, diabetes or abuse were excluded. The outcome measure was treatment at the emergency department after accident as a car driver. RESULTS: Five patients and four controls had been treated, the rate per 1000 person-years with exposure being 3.4 times higher (CI 0.73-17.15) in the patients than in the control cohort. The difference is significant in one-sided test (P=0.04). CONCLUSION: Drivers with MS were treated more often than healthy controls at a casualty department after having a road traffic accident. However, drastic consequences regarding the patients automobile driving should be avoided until these results have been substantiated by further investigations. PMID- 11886360 TI - The controversy of birth order as a risk factor for epilepsy: a study from Saudi Arabia. AB - PURPOSE: To evaluate the relationship between people with epilepsy and birth order. METHODS: A case-control study of 336 epileptics, 15 years and above, and their 1961 full siblings. The data was analysed by birth order and then stratified by sibship size. The 95% confidence interval (CI) for each odds ratio (OR) was obtained. ORs were calculated in the 259 probands in whom the seizure and epileptic syndrome were classifiable against their corresponding 1313 siblings. RESULTS: The OR in birth order 1 is 2.08 (1.6-2.8) on comparing probands to their unaffected siblings. In birth order 3 the OR was 1.64 (1.2-2.2) and ORs declined as birth order increased. The chi-square test for the decline was significant P < 0.05. OR in birth >2 in probands against unaffected sibs was 0.42 (0.2-0.62) in partial seizures and 0.27 (0.17-0.43) in the cryptogenic category, 86% of whom had partial seizures. CONCLUSION: In spite of some limitations in the study it seemed that there is a significant association between low birth order and the risk of epilepsy when all cases were computed together. The cryptogenic type showed the clearest association between low birth order and the likelihood of epilepsy. PMID- 11886361 TI - Silent cerebral infarction and cognitive function in middle-aged neurologically healthy subjects. AB - We sought to clarify whether apparently silent cerebral infarcts and periventricular hyperintensities are associated with depressed cognitive function in middle-aged subjects. Subjects were 84 middle-aged neurologically normal adults who wished to undergo a screening examination of the brain. We performed magnetic resonance imaging (MRI) of the brain and neuropsychologic tests in all subjects. Silent cerebral infarcts and periventricular hyperintensities, respectively, were detected in 21 and 14 of 84 subjects. Mini-mental state (MMS) and Raven's colored progressive matrices (RCPM) scores were significantly lower in subjects with than without silent cerebral infarcts. By two-factor analysis of variance, MMS score was affected by silent cerebral infarcts or periventricular hyperintensities, with interactions between the two lesion types (P < 0.05). Silent cerebral infarcts may be an independent factor in the pathogenesis of intellectual dysfunction, but truly independent analysis is difficult because many subjects with silent cerebral infarcts also have periventricular hyperintensities. PMID- 11886362 TI - Dynamics of LDL oxidation in ischemic stroke patients. AB - OBJECTIVE: Oxidative modification of human low density lipoprotein (LDL) plays an important role in the development of atherosclerosis. The aim of this study was to evaluate the oxidative modification of LDL in the group of patients with ischemic stroke. MATERIAL AND METHODS: In the group of 43 patients 3 months after ischemic stroke and in the age and sex-matched control group, the kinetics of LDL oxidation and level of vitamin E were estimated. The susceptibility of LDL to oxidation was evaluated in isolated LDL exposed to in vitro oxidation. In 26 patients, after diet change, clinical and laboratory investigations were repeated 9 months later. RESULTS: In the patient group, susceptibility of LDL to oxidation was enhanced, lag phase was significantly shorter in comparison with the control group. After a change in diet, significant elongation of the lag phase was observed. CONCLUSION: Diet change improves LDL resistance to oxidation and may influence prognosis in stroke patients. PMID- 11886363 TI - Classic risk factors, hypercoagulability and migraine in young women with cerebral lacunar infarctions. AB - OBJECTIVE: Lacunar cerebral infarctions (LACI) in young women is a rare condition which pathogenesis is still not fully recognized. We explored the presence of classic risk factors, hypercoagulability and migraine in young women with LACI. METHODS: Charts of 192 consecutive premenopausal women suffering cerebrovascular insult [125 (65%) haemorhagic, 58 (30%) ischaemic and 9 (5%) unclassified] during a period of 5 years were reviewed. Sixteen out of 58 (27%) patients with ischaemic stroke were identified to have LACI and included in a study. RESULTS: Ten and seven out of 16 LACI women had at least one classical risk factor (hypertension, hyperlipidaemia, smoking or oral contraceptives) or migraine, respectively. LACI patients had slight hypercoagulable state indicated by shorter thrombin and thromboplastin times, higher fibrinogen and higher t-PA antigen than 47 age matched controls (all P < 0.05). In addition in LACI patients with migraine the trend toward more pronounced hypercoagulable state in comparison to LACI patients without migraine was found. The combination of migraine, at least one classic risk factor and hypercoagulability was present in 5/16 (31.25%) of patients. CONCLUSIONS: The combination of slightly to moderately expressed classic risk factors, hypercoagulability and migraine might be a risk profile for LACI in young women. Further studies are needed to clarify risk profile, rather than isolated risk factors, for LACI in a specific group as young women are. PMID- 11886364 TI - L-Dopa decreases cutaneous nociceptive inhibition of motor activity in Parkinson's disease. AB - OBJECTIVES: To estimate changes in motor inhibitory mechanisms at the spinal level in Parkinson's disease (PD) patients by measuring cutaneous silent responses to nociceptive stimuli in the course of L-Dopa therapy. MATERIALS AND METHODS: Fourteen patients with idiopathic PD (Group 1) and 13 patients with other forms of parkinsonism (Group 2) participated in the study. The cutaneous silent period (CSP) from the hand and clinical scores (UPDRS, part III) were measured "off" therapy (T0), after a single dose of L-Dopa (T1) and 3 months after the beginning of L-Dopa daily therapy (T2). RESULTS: At T0 the duration of the CSP was significantly prolonged in Group 1 and Group 2. At T1 and T2 the mean duration of the CSP significantly decreased in Group 1 (P < 0.05) and a significant correlation was found between the shortening of the CSP and the improvement of rigidity and bradikynesia in the upper limb. CONCLUSIONS: Our findings show that L-Dopa decreases the cutaneous nociceptive inhibition of motor activity in PD patients. CSP may be useful to assess L-Dopa responsiveness during the clinical course of PD. PMID- 11886365 TI - Autosomal dominant cerebellar ataxias in ethnic Bengalees in West Bengal - an Eastern Indian state. AB - BACKGROUND: Phenotypic and genotypic patterns of a hereditary disease in a large multiethnic country like India need to be studied in relation to geographical location and ethnicity of the population. The few reported studies from India on dominant ataxias (ADCA) have mostly been conducted on multiethnic populations and hence may not reflect the patterns observed in specific ethnic groups or geographical locations. The present study attempted to look into the patterns of ADCA amongst ethnic Bengalee patients hailing from the eastern Indian state of West Bengal. MATERIAL AND METHODS: Between mid-1996 and mid-2000, in a clinic based study, 37 cases (from 14 families) with ADCA were studied. This included 33 affected and four asymptomatic members with abnormal physical signs. Genotypic analyses were performed on more than one affected member from each family. Clinical, neuroradiological and electrophysiological aspects were studied. OBSERVATIONS: Genotype analysis revealed: two families with SCA-1,4 families with SCA2,5 families with SCA3 and three families with undetermined genotype. Of the latter, phenotypically 2 were of ADCA 1 and one of ADCA 2 type. No clear preponderance of one particular genotype over another was observed. We noted significant intra- and interfamily variations in phenotype within the same genotype form as well as overlapping of clinical signs between different genotypes. Slow saccadic eye movements and peripheral neuropathy were not seen consistently in our ethnic Bengalee subjects with SCA2 genotypes. Similarly, extrapyramidal features, ophthalmoplegias and distal amyotrophy were seen in some but not in all families with SCA3 mutation. A peculiar form of abduction lag during slow pursuit movement of eyes was observed in an asymptomatic girl in an SCA3 family. CONCLUSIONS: Although SCA2 has been claimed to be the commonest form of ADCA in India, this does not appear to be so in our ethnic Bengalee subjects. Phenotypic expression of the genotype also appears to be variable amongst families and individuals. Hence, phenotypic expression appears to be an inconsistent marker of the SCA genotype in our patients. PMID- 11886366 TI - Parasomnias decline during pregnancy. AB - OBJECTIVES: A survey of the effects of pregnancy on parasomnias. MATERIAL AND METHODS: In an area of a central hospital and the maternity care units in the nearby rural community, women were interviewed during and after their pregnancy with a series of five questionnaires to assess the frequency of their parasomnias. The first questionnaire covered the 3 months before becoming pregnant, the next three the trimesters of pregnancy and the last one the 3 months after delivery. Altogether 325 mothers filled all the five questionnaires and constitute the study group. RESULTS: The total number of parasomnias declined (P < 0.001) during pregnancy and even more among the primiparas than among the multiparas (difference until third trimester, P=0.02). Among various parasomnias reported, sleep talking and sleepwalking decreased from the prepregnant period to the second trimester (22.8 vs 12.6%, change P=0.003), and the reported sleep starts also diminished from the prepregnant time to the first trimester (78.5 vs 63.1%, P < 0.001), but these phenomena did not change further during the follow up. Altogether 55.7% of the women reported having nightmares 3 months before the pregnancy, and 47.7, 49.5, 41.2 and 40.3% (change from the prepregnant period, P < 0.001), respectively, at first, second and third trimester and after the delivery. Reported hypnagogic hallucinations decreased from the prepregnant time to the first trimester (9.8 vs 6.5%, P=0.027), but returned thereafter to the previous level. During the prepregnant period, 25.8% of the women reported bruxism and only 19.9% during the first trimester (P=0.009). Though the prevalence of sleep paralysis decreased during the first trimester of pregnancy, it was the only parasomnia that increased during later pregnancy (from 5.7 to 13.3% in the second trimester, P < 0.013). CONCLUSIONS: The reported frequency of most parasomnias decreases during pregnancy and even more in primiparas than multiparas. PMID- 11886367 TI - History of allergic disorders in common neurologic diseases in Japanese patients. AB - OBJECTIVE: To clarify the association between past and present history of allergic disorders and neurologic diseases. METHODS: The past and present history of common allergic disorders together with family history was prospectively studied in all out-patients at the Department of Neurology at Kyushu University Hospital from March 1998 to February 2000. RESULTS: Among 3113 out-patients, 2152 (69.1%) completed a questionnaire. Myelitis showed a statistically significant increase of concomitant atopic dermatitis (P=0.006) and concomitant and past atopic dermatitis (P=0.014), as compared with neurologically healthy controls. Moreover, patients with lower motoneuron disease (LMND) had a statistically significant increase of past and concomitant asthma (P=0.007). None of the other common neurologic diseases showed any increase of allergic disorders when compared with controls. CONCLUSIONS: The present study supports the significant association between allergic disorders and such spinal cord diseases as myelitis and LMND in Japanese patients. PMID- 11886368 TI - Diagnostic value of electrical stimulation of lumbosacral roots in lumbar spinal stenosis. AB - OBJECTIVES: This paper compares the diagnostic sensitivity of two tests in lumbar spinal stenosis (LSS): lumbosacral root stimulation with needle electrodes and needle electromyograph (EMG). MATERIAL AND METHODS: Twenty patients with LSS were assigned to two groups: Patients with 'neurogenic intermittent claudication' (NIC) only (n=11), and patients with 'neurological signs' (n=9). Ten normal subjects were also examined. The effects of direct stimulation of the lumbosacral roots and conventional EMG recorded from important muscles [rectus femoris (RF): L4, tibialis anterior (TA): L5, soleus muscle (SOL): S1], were compared with each other and correlated with their respective clinical findings and radiological images. RESULTS: Needle EMG and nerve conduction study revealed pathology in 15/20 patients, and electrical stimulation of the roots in 17/20 patients. Agreement in radiological findings with electrical stimulation of the roots and EMG was found in 12 patients. The other patients were harmonic with radiological findings either in EMG or in electrical stimulation of the roots. CONCLUSIONS: Electrical root stimulation revealed more abnormalities in patients with LSS in comparison with needle EMG. However, both methods seemed to complement each other to show additional pathology in a given patient. PMID- 11886369 TI - Palinopsia and perilesional hyperperfusion following subcortical hemorrhage. AB - We report a patient who exhibited transient palinopsia and visual hallucinations. Disturbances initially included an auditory component and increasingly were localized to the left visual field. These events occurred during recovery from a right subcortical hematoma with left homonymous hemianopia. Single-photon emission computed tomography (SPECT) demonstrated extensive perilesional hyperperfusion involving parts of the right parietal, temporal, and occipital cortex. Perilesional hyperperfusion disappeared as the visual abnormalities diminished. We believe that excitatory neuronal activation in perilesional cortex during recovery contributed importantly to the transient abnormal perceptions. PMID- 11886370 TI - Is there a lamotrigine withdrawal syndrome? AB - OBJECTIVES: To report a peculiar observation of a patient who developed a psychomotor inhibition state after a rapid cessation of lamotrigine (LTG). RESULTS: This man was referred to us at the age of 26 years for presurgical evaluation. His treatment [valproate (VPA), 1200 mg/day and LTG, 200 mg/day] was quickly decreased and discontinued after 4 days in order to record seizures. Because LTG was ineffective on seizures control, it was decided to stop it definitively. After a few days, he became anhedonic. He had a tremor, a slight tachycardia and an important hyperhydrosis of the hands. He was considered as having a withdrawal reaction to LTG which was confirmed by spontaneous resolution after a few days. CONCLUSIONS: Withdrawal syndrome caused by anti-epileptic drugs has been rarely reported. However, in our personal experience of patients monitored for epilepsy surgery, many patients complained of minor reactions when the treatments were quickly decreased. Severe reactions are exceptional and may be explained in this case by the pharmacodynamic effects of LTG. It has indeed been suggested that LTG could have psychostimulant and antidepressive effects. PMID- 11886371 TI - Stroke after internal jugular venous cannulation. AB - OBJECTIVE: To alert clinicians to the stroke risk associated with carotid artery injury secondary to attempted internal jugular venous (IJV) cannulation. METHODS: Case reports and review of the literature. RESULTS: Four patients developed a stroke following carotid artery (CA) injury during attempted IJV cannulation using the landmark technique. In all cases the arterial puncture was detected immediately and firm pressure applied for several minutes. In three cases there was evidence of intimal injury and thrombus formation. Two strokes were delayed by more than 24 h. One patient died. A review of studies describing 4487 IJV line insertion attempts using the landmark technique reveals that 5.9% of attempts are associated with CA injury. CONCLUSION: Cannulation of the IJV using visible and palpable landmarks is associated with a risk of stroke. Arterial injury and stroke should be mentioned when consent is obtained for cannulation. Consideration should be given to a reduction of the arterial injury risk by using ultrasound guidance during line insertion. PMID- 11886372 TI - Visual evoked potentials and magnetic resonance imagings of Tolosa-Hunt syndrome before and after steroid therapy. PMID- 11886374 TI - Stomatocytic elliptocytosis and 'neutrophil drumsticks' as a marker of stem cell engraftment. PMID- 11886375 TI - Renal Hodgkin's disease. PMID- 11886376 TI - Sideroblastic anaemias. PMID- 11886377 TI - Class III receptor tyrosine kinases: role in leukaemogenesis. PMID- 11886378 TI - A highly specific and sensitive fluorescence in situ hybridization assay for the detection of t(4;11)(q21;q23) and concurrent submicroscopic deletions in acute leukaemias. AB - The translocation t(4;11)(q21;q23) is one of the most frequent 11q23 abnormalities associated with infant leukaemia as well as topoisomerase inhibitor induced secondary leukaemias. On the molecular level, the MLL gene on 11q23 is fused to the AF4 gene in the 4q21 region, resulting in a chimaeric MLL/AF4 fusion transcript. These particular chromosome rearrangements are generally considered to be associated with poor prognosis, and therefore accurate detection at diagnosis is of clinical significance. In this study we developed a highly specific dual-colour fluorescence in situ hybridization (FISH) assay for the detection of the t(4;11) and demonstrate its usefulness for interphase molecular cytogenetics. In our approach, differentially labelled genomic clones that span the breakpoint cluster regions of both genes involved in the specific translocation were used. Thus, t(4;11)-positive nuclei will display two fusion signals and for t(4;11) cases with concurrent 3' MLL deletions only one fusion signal will be displayed. A very low false-positive value of less than 0.1% was obtained for interphase cells with two fusion signals. In contrast, in cases with 3' MLL deletions that display only one fusion signal, the rate of false-positive nuclei was 10.4%. This FISH assay enables the screening of larger series of patients with haematological diseases for t(4;11) translocations and allows the unambiguous detection of associated cryptic deletions. PMID- 11886379 TI - Expression of the signal transduction molecule zeta in peripheral and tumour associated lymphocytes in Hodgkin's disease in relation to the Epstein-Barr virus status of the tumour cells. AB - We investigated whether the described immune evasion of Epstein-Barr virus (EBV) infected malignant Hodgkin and Reed-Sternberg (HRS) cells in Hodgkin's disease (HD) is paralleled by a disturbed expression of the signal transduction molecule zeta associated with CD3 and CD16 in tumour-associated T lymphocytes (TAL). Flow cytometric analysis revealed a significantly lower zeta expression in CD3+/4+, CD3+/8+ and CD16+ patient peripheral blood lymphocytes (PBL; n = 10) compared with normal donor PBLs (n = 11). When patient PBLs were compared with the corresponding TAL, the latter showed a significantly higher (CD3+/4+) or equal (CD3+/8+) zeta expression. The EBV status of the tumours did not correlate with zeta expression in the TAL. Immunohistochemical staining revealed zeta-positive lymphocytes among the adjacent bystander cells of the HRS cells in all analysed tumours (n = 8), irrespective of tumour EBV status. In conclusion, these results do not support downregulation of zeta in TAL as a critical mechanism contributing specifically to the immune escape of EBV+ HRS cells. PMID- 11886380 TI - Multidrug resistance mechanisms in chronic lymphocytic leukaemia. AB - We evaluated the presence of P-glycoprotein (P-gp)-170, multidrug resistance protein (MRP), lung resistance protein (LRP)-56 and Bcl-2 in CD19-positive cells from 100 cases of chronic lymphocytic leukaemia (CLL). P-gp-170 was found in 73% of the CLL cases with no significant difference regarding stage or previous treatment. LRP-56 protein was homogeneously distributed with no differences for stage or treatment. MRP protein was detected at a low level of expression in 49.4% of CLL patients with no differences for stage or treatment. Bcl-2 protein was expressed at a high level in all CLL patients and higher levels were found in the advanced stage. This leads us to conclude that P-gp, MRP, LRP-56 and Bcl-2 are frequently expressed in CLL. P-gp, MRP and LRP are not correlated to stage or previous treatment. Bcl-2 is higher in advanced-stage patients. The clinical and biological significance of these zMDR mechanisms in CLL remains to be fully explained. PMID- 11886381 TI - High-dose chlorambucil for the treatment of chronic lymphocytic leukaemia and low grade non-Hodgkin's lymphoma. AB - Chlorambucil has been used for many years for the treatment of low-grade B-cell lymphoproliferative disorders, including chronic lymphocytic leukaemia and low grade non-Hodgkin's lymphoma. There is evidence in the literature that increasing the dose of chlorambucil produces better results than 'standard' doses in terms of response rates and overall survival. There is also evidence that this approach may be at least as effective as the use of fludarabine, as well as being very much less expensive. We describe a high-dose chlorambucil (HDC) regimen, which involves a sustained but intermittent dose of chlorambucil, i.e. 30 mg/d for 4 d per week for 4 weeks, followed by a further four courses at fortnightly intervals for 8 weeks (a total of eight 4-d courses) given as a single drug over an initial 12-week period. The outcome of treatment in previously treated and untreated patients was excellent, with a median time to treatment failure of 33 months for the patient cohort overall and for previously treated and chemotherapy-naive patients of 13 and 104 months respectively. In patients previously treated with fludarabine, 78% had a response. Autoimmune haemolytic anaemia was reversed in one patient. Toxicity, both haematological and other, was minimal. We propose that escalated-dose chlorambucil regimens should be compared with fludarabine in randomized controlled trials, rather than 'standard' lower dose protocols. PMID- 11886382 TI - Interleukin 10 abolishes the growth inhibitory effects of all-trans retinoic acid on human myeloma cells. AB - Recently, it was disclosed that all-trans retinoic acid (ATRA) inhibits myeloma cell growth by downregulating the interleukin 6 (IL-6)/IL-6 receptor (IL-6R) auto/paracrine loop, and upregulating p21/Cip1 cyclin-dependent kinase inhibitor (CDK-I), thereby inducing apoptosis with a decrease in Bcl-2 protein expression. To elucidate and generalize the effects of ATRA on the proliferation and cellular biology of myeloma cells, 12 human myeloma cell lines established in our laboratory were utilized. Two out of the 12 lines showed enhanced growth on supplementation of ATRA and were characterized by IL-10 production, downregulation of membrane Fas and reduced upregulation of p21/Cip1 CDK-I message. These characteristics may prove important for the clinical use of ATRA and should be considered before starting ATRA therapy for myeloma. PMID- 11886383 TI - Clinical significance of vascular endothelial growth factor and hepatocyte growth factor in multiple myeloma. AB - Angiogenesis is a crucial process in the progression of multiple myeloma (MM). Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) are multifunctional cytokines that potently stimulate angiogenesis including tumour neovascularization. Serum levels of VEGF and HGF were measured in 52 patients with MM by enzyme-linked immunosorbent assay (ELISA). Serum levels of VEGF and HGF were elevated in MM patients compared with healthy controls (VEGF: mean 0.31 ng/ml and 0.08 ng/ml respectively, P < 0.01; HGF: mean 2.17 ng/ml and 0.45 ng/ml, respectively, P < 0.001). In serial samples taken after chemotherapy, serum VEGF and HGF levels were correlated with M-protein levels. Serum levels of VEGF were higher in patients with extramedullary plasmacytomas than in patients without them (P < 0.05). They were also significantly higher in a group of patients who showed poor response to chemotherapy (P < 0.01). Serum levels of HGF were higher in patients with complications such as anaemia, hypercalcaemia and amyloidosis than in patients without these complications (P < 0.01, P < 0.05, P < 0.05 respectively). Both serum VEGF and HGF levels were significant predictors of mortality (P = 0.01, P = 0.02, respectively, log-rank test). The present study demonstrated that serum levels of VEGF and HGF are significantly elevated and dependent on the severity of MM, suggesting that measurement of VEGF and HGF may be useful for assessing disease progression and for predicting the response to chemotherapy in MM patients. PMID- 11886384 TI - Clinical evaluation of a polymerase chain reaction assay to detect Aspergillus species in bronchoalveolar lavage samples of neutropenic patients. AB - The increasing incidence of invasive aspergillosis, a life-threatening infection in immunocompromised patients, emphasizes the need to improve the currently limited diagnostic tools. Using a recently developed two-step polymerase chain reaction (PCR) assay to detect 10 fg of Aspergillus DNA, corresponding to 1-5 colony-forming units (CFU)/ml of spiked samples in vitro, we prospectively examined 197 bronchoalveolar lavage (BAL) samples from 176 subjects, including 141 neutropenic, febrile patients with lung infiltrates, at risk for invasive fungal disease. Underlying diseases of these patients were haematological malignancies; 93 patients suffered from acute leukaemias. Thirty-one of these immunocompromised patients (17.6%) were PCR positive, correlating with positive BAL culture, positive histology from lung surgery or from autopsy, positive computerized tomography scans or positive galactomannan enzyme-linked immunosorbent assay. Six patients (4.3%) of this group had positive PCR results without any correlation to clinical or other diagnostic data, probably owing to contamination of the samples by ubiquitous Aspergillus spores. The samples of two patients (1.4%) with a subsequent histologically proven mould infection were PCR negative. All 102 immunocompromised patients (72.3%) with a negative PCR showed no evidence of invasive fungal disease. From 35 patients without immunodeficiency, four (11.4%) showed positive results, without evidence of invasive or non-invasive pulmonary aspergillosis. In this haematological population, the sensitivity and specificity values of the test reached 93.9% and 94.4%, the positive predictive value 83.8%, the negative predictive value 98.1%. Our data support the considerable clinical value of this PCR assay for confirming and improving diagnosis of pulmonary aspergillosis in high-risk patients. PMID- 11886387 TI - Outbreak of vancomycin-resistant Enterococcus faecium in a haematology unit: risk factor assessment and successful control of the epidemic. AB - We describe an outbreak of vancomycin-resistant Enterococcus faecium (VRE) on the haematology ward of a Dutch university hospital. After the occurrence of three consecutive cases of bacteraemia with VRE, strains were genotyped and found to be identical. During the next 4 months an intensive surveillance programme identified 21 additional patients to be colonized with VRE, while two more patients developed bacteraemia. A case-control study was carried out to identify risk factors for VRE acquisition. In comparison with VRE-negative control patients (n=49), cases (n=24) had a longer stay on the ward during the year preceding the outbreak (25.8 versus 10.1 d, P=0.02), more cases with acute myeloid leukaemia [11 versus 4, odds ratio (OR) 9.5, 95% confidence interval (CI95) 2.4-32.2] and higher grades of mucositis (P=0.03). Logistic regression analysis identified antibiotic use within 1 month before admission (OR 13.0, CI95 2.1-80.5, P=0.006) and low albumin levels at baseline (OR 1.2, CI95 1.1-1.3, P=0.02) to be independent risk factors. Four patients with VRE-bacteraemia were successfully treated with quinupristin/dalfopristin (Synercid). Control of the outbreak was achieved by step-wise implementation of intensive infection control measures, which included the cohorting of patients, allocation of nurses and reinforcement of hand hygiene. PMID- 11886386 TI - Excessive apoptosis, increased phagocytosis, nuclear inclusion bodies and cylindrical confronting cisternae in bone marrow biopsies of myelodysplastic syndrome patients. AB - Transmission electron microscopic (TEM) studies have not been reported in bone marrow (BM) biopsies of patients with myelodysplastic syndromes (MDS) owing to failure to overcome the technical impediment of maintaining ultrastructural detail in decalcified tissue. Using a modified technique to physically separate pieces of bone from marrow tissue under a dissecting microscope, and embedding the material directly for TEM, ultrastructural studies were performed in 15 MDS patients and four normal BM biopsies. Biopsy tissue was also used to initiate long-term in vitro cultures and 12-week plates were sacrificed for TEM analysis. Features noted in freshly obtained decorticated tissue included an excessive apoptosis in both haematopoietic and stromal cells, ringed sideroblasts with iron laden mitochondria and highly active, enormously increased phagocytosis. In addition, type IV nuclear inclusion body variants (NIB-v) and confronting cylindrical cisternae (CCC) were readily identified in up to 40% of stromal cells in vivo, providing an important footprint of a possible infectious agent in the pathology of MDS. Cultured stromal cells did not show excessive apoptosis and only 2-4% fibroblasts showed the presence of NIB-v or CCC, underscoring the artificial nature of ex vivo systems. We conclude that ultrastructure studies using decorticated tissue can be a powerful tool to investigate the biology and aetiology of a variety of haematopoietic disorders as it enables the direct examination of BM biopsies with their intimate stromal parenchymal cell associations preserved intact. PMID- 11886385 TI - Typical essential thrombocythaemia does not express bcr-abelson fusion transcript. AB - Essential thrombocythaemia (ET) is a chronic myeloproliferative disorder (MPD) characterized by an elevated platelet count and no identifiable underlying primary cause. According to the diagnostic criteria of the Polycythemia Vera Study Group (PVSG), ET lacks features diagnostic for other MPDs, including the Philadelphia chromosome (Ph) or bcr-abl rearrangement. Recently, some authors have reported bcr-abl transcript positivity in ET patients, but these findings remain controversial. The aim of this study was to investigate whether the bcr abl transcript could be found in ET patients and to verify the hypothesis of a new ET variant. ET patients (n = 121) with a median age at diagnosis of 55 years were enrolled. The bcr-abl transcript status was examined by multiplex reverse transcription-polymerase chain reaction. Only two cases were positive for bcr abl, one of which had the Ph at diagnosis. The positive bcr-abl transcript was associated, in both cases, with mild basophilia at diagnosis. After a median follow-up of 43 months (0-309 months), two patients in the bcr-abl-negative group developed Ph and bcr-abl-negative acute myeloid leukaemia (AML). In contrast, one of the two patients in the bcr-abl-positive group died from AML 13 years after diagnosis. In conclusion, our data on a large group of patients shows the rarity of the bcr-abl transcript in well-established ET. However, a subset of patients with apparent ET and basophilia may express the transcript and may constitute a novel entity intermediate between chronic myeloid leukaemia (CML) and typical ET. A prospective study is warranted in order to define better the clinical and biological characteristics of bcr-abl-expressing ET. PMID- 11886388 TI - High multidrug resistance protein activity in acute myeloid leukaemias is associated with poor response to chemotherapy and reduced patient survival. AB - Multidrug resistance protein (MRP) activity was investigated in 44 newly diagnosed acute myeloid leukaemia (AML) patients using a functional assay based on efflux of carboxy-2',7'-dichlorofluorescein, an anionic dye handled by both MRP1 and MRP2. Elevated MRP transport was detected in 29% of cases, but was not significantly correlated with sex, age, white blood cell count at diagnosis or karyotype. In contrast, it was associated with secondary AML (P = 0.002), CD34 positivity (P = 0.041) and P-glycoprotein activity (P = 0.01). There was a lower rate of complete remission in MRP-positive patients versus MRP-negative patients (23% versus 81%; P = 0.001); overall survival was also better for MRP-negative patients (P = 0.004). These data indicate a probable role for MRP activity in the clinical outcome of AML. PMID- 11886389 TI - Genotyping of human platelet antigen-1 by gene amplification and labelling in one system and automated fluorescence correlation spectroscopy. AB - Genotyping of human platelet antigen-1 (HPA-1) is required for the diagnosis and appropriate therapy of alloimmunization. Recently, the HPA-1 polymorphism has been identified as an inherited risk factor for thrombosis. Most currently used methods for HPA-1 genotyping have the disadvantage of time-consuming post polymerase chain reaction (PCR) processes such as ligation (oligonucleotide ligation assay), restriction enzyme digestion (allele-specific restriction enzyme analysis) and electrophoresis (single-strand conformation polymorphism). We present a novel method for HPA-1 genotyping based on a homogeneous PCR strategy (GALIOS, gene amplification and labelling in one system) combined with automated fluorescence correlation spectroscopy (FCS). The PCR uses one pair of gene specific amplification primers and two allele-specific, semi-nested labelling primers. The allele-specific labelling primers differ in a single nucleotide (T for HPA-1a/1a, C for HPA-1b/1b) and are coupled to different fluorescent dyes. The quantities of generated fluorescent PCR products are analysed by FCS at 543 nm and 633 nm excitation wavelength respectively. The genotypes determined using this method were in 100% concordance with the results obtained by allele-specific restriction analysis (n = 380 samples). The assay was validated for specificity, reliability and the dynamic range. This innovative method of rapid HPA-1 genotyping offers a specific and robust system, which is applicable for routine HPA-1 genotyping. PMID- 11886390 TI - European Concerted Action on Anticoagulation (ECAA): multicentre international sensitivity index calibration of two types of point-of-care prothrombin time monitor systems. AB - A multicentre modified World Health Organization (WHO)-type international sensitivity index (ISI) calibration has been performed at 10 European Concerted Action on Anticoagulation (ECAA) national laboratories using non-citrated whole blood on two point-of-care test (POCT) prothrombin time (PT) monitor systems, CoaguChek Mini and TAS PT-NC, using single lots of test cards/strips. The relevant species (human and rabbit) WHO international reference preparations (IRPs) were tested with the manual PT technique on citrated plasma from the same blood donations. The ISI was calculated from the slope of the orthogonal regression line relating log PT (POCT) to log PT (IRP). The mean ISI of the CoaguChek Mini system was 1.75 and 1.13 with the prothrombin time non-citrated Thrombolytic Assessment System (TAS PT-NC). With the CoaguChek Mini system, seven out of 10 calibrations exceeded the current 3% WHO recommended limit for the coefficient of variation (CV) of the slope with conventional PT testing, whereas with the TAS PT-NC system, it was eight out of 10. All the POCT calibrations had a CV of the slope <5%. It is suggested that this level of precision be adopted as the limit of acceptability of calibration of these monitor systems. In these circumstances, the modified WHO-type ISI calibration appeared to be satisfactory for the POCT whole-blood monitors. PMID- 11886391 TI - Hormonal replacement therapy, prothrombotic mutations and the risk of venous thrombosis. AB - Hormone replacement therapy (HRT) increases the risk of venous thrombosis. We investigated whether this risk is affected by carriership of hereditary prothrombotic abnormalities. Therefore, we determined the two most common prothrombotic mutations, factor V Leiden and prothrombin 20210A in women who participated in a case-control study on venous thrombosis. Relative risks were expressed as odds ratios (OR) with 95% confidence intervals (CI95). Among 77 women aged 45-64 years with a first venous thrombosis, 51% were receiving HRT at the time of thrombosis, compared with 24% of control women (OR = 3.3, CI95 1.8 5.8). Among the patients, 23% had a prothrombotic defect, versus 7% among the control women (OR = 3.8, CI95 1.7-8.5). Women who had factor V Leiden and used HRT had a 15-fold increased risk (OR = 15.5, CI95 3.1-77), which exceeded the expected joint odds ratio of 6.1 (under an additive model). We conclude that the thrombotic risk of HRT may particularly affect women with prothrombotic mutations. Efforts to avoid HRT in women with increased risk of thrombosis are advisable. PMID- 11886392 TI - Pipobroman is safe and effective treatment for patients with essential thrombocythaemia at high risk of thrombosis. AB - Essential thrombocythaemia (ET) is a disease associated with an elevated risk of thrombosis. This study evaluated the efficacy and safety of pipobroman (PB) in the long-term control of ET patients who had, at diagnosis, one or more of the following currently known risk factors for thrombosis or haemorrhage (high-risk patients): age > 60 years, history of thrombosis or haemorrhage, platelets >1000 x 10(9)/l. From 1978 to 2000, with a median follow-up of 10 years, 118 previously untreated high-risk ET patients (median age 62 years, range 25-82), were treated with PB at the starting dose of 0.8-1 mg/kg/d. All patients reached a platelet count <600 x 10(9)/l and 91% achieved a platelet count <400 x 10(9)/l. During follow-up, 13 patients had thrombosis, with a 10-year cumulative risk of 14%. Acute myeloid leukaemia, myelofibrosis and solid tumours occurred in three, two and seven patients with a 10-year cumulative risk of 3%, 2% and 7% respectively. Actuarial survival at 20 years was 64% and the standardized mortality ratio was 1.1 (95% CI: 0.7-1.7), not statistically different from the general population (P = 0.54). Age was associated with a higher risk of death (P = 0.00009) and thrombosis (P = 0.003). The duration of PB treatment did not correlate with the occurrence of second malignancies. This study, with a median follow-up of 10 years, demonstrates that pipobroman is effective and well tolerated. The low cumulative 10-year risk of thrombosis, leukaemia and solid tumours indicates that pipobroman is an adequate treatment for patients with high risk ET. PMID- 11886393 TI - Reduced transforming growth factor-beta1 production by mononuclear cells from patients with active chronic idiopathic thrombocytopenic purpura. AB - Chronic idiopathic thrombocytopenic purpura (ITP) is an autoimmune disorder in which activated T-helper (Th) cells and different Th-cell cytokines might play an important role. We have recently reported that chronic ITP patients in remission had elevated plasma levels of the Th3 cytokine transforming growth factor-beta1 (TGF-beta1), possibly as a part of a bystander immune suppression. In the present study we found that, in ITP patients with active disease [platelet count (plc) < 50 x 10(9)/l], mitogen-stimulated peripheral blood mononuclear cells (PBMC) had a significantly reduced production of TGF-beta1 (444 +/- 178 pg/ml; n = 6) compared with patients with plc 50-150 x 10(9)/l (1293 +/- 374 pg/ml; n = 9; P < 0.05), patients with plc >150 x 10(9)/l (1894 +/- 244 pg/ml; n =12; P <0.005) and healthy controls (1698 +/- 241 pg/ml; n = 10; P < 0.01). Nineteen per cent of ITP patients expressed a platelet-induced PBMC proliferation. Surprisingly, 22% of the ITP patients had a PBMC proliferation below the normal range, i.e. a suppressed proliferation in the presence of platelets; five of these six patients had active disease. In summary, this study demonstrated that chronic ITP patients with active disease had reduced PBMC production of the Th3 cytokine TGF-beta1. This result gives further support to the theory that chronic ITP in active phase is associated with a downregulated Th3-response. PMID- 11886394 TI - Anticoagulant effects of a synthetic peptide containing residues Thr-2253-Gln 2270 within factor VIII C2 domain that selectively inhibits factor Xa-catalysed factor VIII activation. AB - Factor VIII (FVIII), an essential cofactor that accelerates the generation of factor Xa (FXa) in the tenase complex, is activated by proteolytic cleavage by thrombin or FXa. A strong relationship has been reported between high levels of FVIII activity and thrombosis. We have demonstrated previously that an anti-FVIII C2 antibody (ESH8) with a Val-2248-Gly-2285 epitope inhibited FXa-catalysed FVIII activation, and that a synthetic peptide designated EP-2 (residues 2253-2270) blocked C2 domain binding to FXa. We investigated the inhibitory effect of EP-2 on FXa-catalysed FVIII activation and its anticoagulant effect in the blood coagulation system. EP-2 inhibited FXa-catalysed activation in a clotting assay in a dose-dependent manner and reduced FXa generation in a chromogenic assay using FVIII, factor X, factor IXa and phospholipid. The peptide only inhibited FVIII binding to FXa. We also tested the anticoagulant effect of EP-2 in the plasma milieu. The peptide prolonged the activated partial thromboplastin time and activated clotting time in a dose-dependent manner, but not prothrombin time. Our results indicate that EP-2 mediates the anticoagulant effect by specific inhibition of FVIII and FXa interaction in the intrinsic pathway, and that FXa catalysed FVIII activation plays a significant role in blood clotting. The peptide may provide the basis for the development of novel anticoagulant therapy. PMID- 11886395 TI - True anti-anionic phospholipid immunoglobulin M antibodies can exert lupus anticoagulant activity. AB - True (cofactor-independent) anticardiolipin antibodies (aCL) are thought to lack lupus anticoagulant (LA) activity and pathogenic potential. A serum monoclonal immunoglobulin Mlambda (mIgMlambda) with aCL and LA activities found in a man with a splenicIgMlambda+ B-cell lymphoplasmacytic lymphoma (LPL) without thrombotic events has been characterized. LPL-derived hybridoma clones (designated HY-FRO) producing the serum mIgMlambda were obtained. mIgMlambda secreted by HY-FRO grown in protein-free culture medium, like that purified from serum, (i) showed binding, in a cofactor-free system, to solid-phase CL and phosphatidylserine (PS) and to the membrane of PS-expressing cells (apoptotic cells and activated platelets); (ii) failed to bind neutral phospholipids (PL), beta2Glycoprotein, histone, ssDNA, dsDNA, human IgG and umbilical vein endothelial cells. Absorption with apoptotic cells abolished its binding to anionic plate-bound CL and PS. IgMlambda-FRO used poorly mutated VH and Vlambda region genes, with a pattern that was inconsistent with an antigen-driven selection. Basic amino acids were present in the IgH complementarity determining region 3 (CDR3), which can be important for binding to anionic PL. These findings demonstrate unequivocally that true anti-anionic PL IgM antibodies can exert LA and indicate this anti-PL type does not involve thrombophilia. PMID- 11886396 TI - The new ID-heparin/PF4 antibody test for rapid detection of heparin-induced antibodies in comparison with functional and antigenic assays. AB - Heparin-induced thrombocytopenia (HIT) is an immune-mediated complication of heparin treatment. Several in vitro assays are available to detect the causative HIT antibodies: functional assays, usually requiring freshly prepared platelets and immunological tests based on the enzyme-linked immunosorbent assay (ELISA) principle. We compared a new, simple and rapid test based on the ID-microtyping particle agglutination system with 14C-serotonin release assay, heparin-induced platelet activation (HIPA) test and two ELISAs. Sera from 100 confirmed HIT patients, 20 serologically negative suspected HIT patients and 20 healthy blood donors were used. The specificity and sensitivity of the new test was similar to the functional assays. Compared with the ELISAs, specificity was better at the cost of reduced sensitivity. As in all other immunological tests, HIT antibodies against less typical antigens, such as interleukin (IL)-8 or neutrophil activating peptide (NAP) 2 could not be detected. Thus, although the ID Heparin/PF4 antibody test seems to be a quick, reliable and robust test to determine the presence of HIT antibodies, it should still be combined with a functional assay if possible. Evaluation of the test in a prospective setting as well as interlaboratory variation should be assessed as a next step. PMID- 11886397 TI - Effect of nitric oxide modulation on systemic haemodynamics and platelet activation determined by P-selectin expression. AB - Inhibiting platelet and endothelial nitric oxide production favours platelet adhesion and aggregation, and arterial vasoconstriction. This study investigated the effect of NG-nitro-l-arginine methyl ester (L-NAME), a stereospecific inhibitor of nitric oxide synthesis, on P-selectin expression on platelets, platelet-derived microparticles and platelet-leucocyte aggregates, and on soluble P-selectin levels. Twelve healthy male volunteers were infused intravenously with L-NAME and then with a 10% solution of either l- or d-arginine. Blood pressure responses were recorded and whole blood and serum collected at baseline and after each infusion. P-selectin expression was analysed in all samples by flow cytometry. Serum levels of soluble P-selectin were batch analysed using an enzyme linked immunosorbent assay at the end of the study. P-selectin expression on platelets, platelet-derived microparticles and platelet-leucocyte aggregates did not vary significantly from baseline levels following the infusion of L-NAME or l or d-arginine. However, endothelial nitric oxide synthase inhibition caused a marked elevation of arterial blood pressure (P < 0.01) that was restored to pretreatment values by l- but not d-arginine. Serum levels of the soluble form decreased significantly (P = 0.001) following the infusion of l- and d-arginine compared with samples taken at baseline and following L-NAME infusion. In conclusion, inhibition of constitutive nitric oxide synthase in the endothelium and platelets produced significant increases in blood pressure but did not alter platelet membrane expression of P-selectin. PMID- 11886398 TI - Lack of multimer organization of von Willebrand factor in an acquired von Willebrand syndrome. AB - We report a case of acquired von Willebrand syndrome (AVWS) in a 20-year-old woman with systemic lupus erythematosus, in whom severe bleeding complications followed kidney biopsy. Coagulation studies demonstrated undetectable levels of ristocetin-induced platelet aggregation (RIPA), von Willebrand factor antigen (VWF:Ag) and VWF ristocetin cofactor activity (VWF:RCo), associated with significantly prolonged bleeding time; unlike type 3 von Willebrand disease (VWD), platelet VWF was reduced but not undetectable. The plasma VWF multimer pattern was characterized by the presence of only two bands, one of low molecular weight (MW) running as the protomer of plasma VWF in normals, the other of abnormally high MW without detectable intermediate multimers; this pattern resembles that of VWF present in endothelial cells. A search for an anti-VWF antibody demonstrated the presence of an inhibitor at high titre. This anti-VWF antibody did not interfere in the interaction of VWF with platelet glycoprotein (GP) Ib through the A1 domain, and did not react with the A2 domain of VWF; instead, it seemed to modify the relative representation of high and low MW VWF multimers released by normal human umbilical vein endothelial cells (HUVEC). After Azathioprine and corticosteroid treatment, the anti-VWF antibody disappeared and the patient's haemostatic profile normalized, except for the platelet VWF content which still remained decreased. We suggest that the anti-VWF antibody present in the AVWS described compromised both circulating VWF levels and their multimeric organization, inducing the maintenance of the multimer structure that VWF normally has before or in the early phase after secretion from endothelial cells. PMID- 11886399 TI - Plasma exchange as a source of protein C for acute onset protein C pathway failure. AB - Severe bacterial sepsis, particularly secondary to meningococcaemia, is a well recognized cause of purpura fulminans resulting from severe acquired protein C (PC) deficiency. Recently, PC and activated protein C (APC) concentrate replacement therapy has been shown to improve outcome in patients with meningococcaemia- associated purpura fulminans and severe sepsis respectively. Despite these impressive findings, PC and APC concentrates are not currently widely available. We describe a 31-year-old patient with pneumococcal septic shock, purpura fulminans (PF) and severe acquired PC deficiency, whom we successfully treated with conventional therapy and high-volume plasma exchange as a source of PC. PMID- 11886400 TI - von Willebrand factor-cleaving protease inhibitor in a patient with human immunodeficiency syndrome-associated thrombotic thrombocytopenic purpura. AB - Antibodies that inhibit von Willebrand Factor (VWF)-cleaving protease activity occur in patients with acute thrombotic thrombocytopenic purpura (TTP) and often persist in the chronic phase. A deficiency of this protease is likely to be responsible for the generation of ultrahigh VWF multimers and influence the formation of intra-arterial platelet aggregates that result in microangiopathic haemolytic anaemia, thrombocytopenia and end in organ failure. This report demonstrates complete deficiency of VWF-cleaving protease and the presence of a concentration-dependent IgG1 inhibitor in the plasma of a patient with acquired immunodeficiency syndrome (AIDS). These data may contribute to understanding the pathophysiology of human immunodeficiency syndrome (HIV)-related TTP. PMID- 11886401 TI - Ex vivo expansion of megakaryocyte progenitor cells from normal bone marrow and peripheral blood and from patients with haematological malignancies. AB - A number of haematological and non-haematological malignancies can be successfully treated using high-dose chemotherapy +/- irradiation followed by haematopoietic progenitor cell transplantation. Post transplant, thrombocytopenia and neutropenia always occur and patients require platelet transfusions. It may be possible to reduce the period of thrombocytopenia by re-infusion of ex vivo expanded megakaryocyte progenitors (MP), derived from the progenitor cell graft. We have investigated the expansion of MP from CD34+ enriched cells from normal bone marrow (NBM) and peripheral blood (PB) and remission BM or PB samples from patients with haematological malignancies. CD34+ cells were cultured in serum free medium supplemented with thrombopoietin (TPO), interleukin 1 (IL-1), IL-6 and stem cell factor (SCF) for 7 d, then cell proliferation was assessed by flow cytometry using lineage-specific markers. It was possible to significantly expand the number of MP cells from all sources. There were no major differences in yields of MP from normal BM or PB, or BM from multiple myeloma and non-Hodgkin's lymphoma patients. However, expansion of MP in acute myeloid leukaemia samples was lower than all other samples and the number of megakaryocyte colony-forming units was reduced. Several cytokine combinations were evaluated to optimize MP expansion from NBM. Equivalent yields of MP were obtained using TPO and one of IL 1, IL-3, granulocyte-macrophage colony-stimulating factor or SCF, suggesting that large cytokine combinations are not necessary for this procedure. It should be possible to scale up the culture conditions described to produce effective MP doses for clinical transplantation. PMID- 11886402 TI - Adoptive therapy with monocyte-derived macrophages in the setting of high-dose chemotherapy and peripheral blood stem cell transplantation. AB - In an attempt to ameliorate chemotherapy-induced side-effects after transplantation of autologous peripheral blood stem cells (PBSCT), we tested the reinfusion of autologous macrophages (MAC) that are known to be potent antimicrobial effector cells and cytokine producers. Ten patients were treated with two sequential cycles of high-dose chemotherapy followed by PBSCT. Before the second cycle of PBSCT, mononuclear cells were harvested, cultured for 8 d in order to induce MAC maturation and reinfused 3 d after PBSCT without clinical problems. However, MAC infusions did not substantially alleviate the toxicity of autologous PBSCT. PMID- 11886403 TI - Second malignancies in patients with essential thrombocythaemia. PMID- 11886404 TI - Veno-occlusive disease after an anti-CD33 therapy (gemtuzumab ozogamicin). PMID- 11886407 TI - Our responsibility in a developing world: from ethics to pragmatism. AB - If development is defined as a process of enhancing human capabilities, that is, to expand choices and opportunities so that each person can lead a life of respect and value, then poverty is the deprivation of these capabilities. Nobel Laureate for Economics, Amartya Sen, states: 'as people who live minus sign in a broad sense minus sign together, we cannot escape the thought that the terrible occurrences that we see around us are quintessentially our problems'. This year's Council Lecture examines issues of individual and institutional responsibility in a developing world. Aspects of development relevant to ophthalmology are discussed and a review of Australian efforts undertaken. With a view to encouraging Fellows to take a more active role in development, it is demonstrated that there are a range of contributions that can be made. Appropriate practice models are explored and a strategy for College involvement presented. PMID- 11886406 TI - Flies and trachoma. PMID- 11886408 TI - Clinical aspects of conjunctival melanoma. AB - Conjunctival melanoma is a rare but important condition encountered in ophthalmology. This paper reviews conjunctival melanoma as a clinical entity, with particular emphasis on differential diagnosis, management and prognostic factors. Relevant references were located through a comprehensive search of articles published between 1980 and early 2001 on Medline, using both the WinSpirs and PubMed platforms. The references of these articles were then checked for further articles of relevance. The condition is uncommon and its presentation variable; therefore, relevant studies were found to suffer from small numbers of subjects. This may be the cause of the confusion that still surrounds the condition and its management. PMID- 11886409 TI - Seasonal variation in trachoma and bush flies in north-western Australian Aboriginal communities. AB - BACKGROUND: Among younger age groups trachoma (Chlamydia trachomatis) has been identified as a major cause of morbidity in Australian Aboriginal communities. North-western Australia has two seasons, referred to as the wet and the dry, and until recently most trachoma screening programmes were conducted during the dry season. This study compared the prevalence of trachoma between three Aboriginal communities, two in the west and one in the east Kimberleys with differences in adult bush fly (Musca vetustissima) populations between the wet and dry seasons. METHODS: All preschool and school-aged children in each community were screened for trachoma in February and July 1996 using the World Health Organization method for clinical assessment of trachoma. Flies were trapped fortnightly from September through to May (inclusive) using a wind-orientated fly trap. RESULTS: Two communities in the west Kimberleys had a significantly higher rate of trachoma during the wet season (14-59% in dry season compared with 46-69% in wet season). One community showed no difference but this was probably due to the reduced re-screening rate. Further-more, it was demonstrated that fly populations are so low during the dry season that they were untrappable; however, populations of bush fly significantly increased during the wet season (ranging from 6 to 146 flies per hectare per month). CONCLUSIONS: If bush fly populations are correlated with increased levels of trachoma, then measures aimed at augmenting public health plans for bush fly control may decrease the cross-infection rate. Additionally, based on the results of this study, wet season trachoma screening trips should be considered. PMID- 11886410 TI - Population-based assessment of refractive error in India: the Andhra Pradesh eye disease study. AB - PURPOSE: To assess the prevalence, distribution, and demographic associations of refractive error in the population of the southern Indian state of Andhra Pradesh. METHODS: From 94 clusters in one urban and three rural areas of Andhra Pradesh, 11 786 persons of all ages were sampled using a stratified, random, cluster, systematic sampling strategy in the Andhra Pradesh Eye Disease Study, a population-based cross-sectional study. A total of 10 293 people underwent an interview and detailed dilated eye examination. Refraction was performed by ophthalmic personnel trained in the study procedures. Objective refraction under cycloplegia was assessed for participants < or = 15 years of age and subjective refraction for those > 15 years of age. Myopia was defined as spherical equivalent worse than -0.50 D and hyperopia as spherical equivalent worse than +0.50 D. RESULTS: In the participants < or = 15 years of age, the prevalence of myopia was 3.19% (95% confidence interval [CI] 2.24-4.13%) and of hyperopia was 62.62% (95% CI 57.10-68.13%). In this age group, myopia increased with increasing age and was more prevalent in the urban study area, and hyperopia prevalence was greater in the participants < 10 years of age. In participants > 15 years of age, the prevalence of myopia was 19.45% (95% CI 17.88-21.02%) and of hyperopia was 8.38% (95% CI 6.91-9.85%). Myopia and hyperopia increased with increasing age. Myopia was more common in males, those with education higher than class 12, those with nuclear cataract, and those living in rural study areas. Hyperopia was more common in females, those with any level of formal education, and those living in the urban area and in the well-off rural study area. CONCLUSIONS: There is significant refractive error in this population. These data on the distribution and associations of refractive error can be useful for the planning of refractive eye-care services. PMID- 11886411 TI - Use of mitomycin C in the treatment of corneal conjunctival intraepithelial neoplasia. AB - PURPOSE: To evaluate the efficacy of topical mitomycin C as a treatment of corneal conjunctival intraepithelial neoplasia. METHODS: An open prospective analysis of 20 cases of corneal conjunctival intraepithelial neoplasia with recurrent disease (17 patients) or refusing surgery (three patients) were treated with topical mitomycin C. Treatment was with mitomycin C eye drops, either 0.02% or 0.04%, four times daily for 1 week followed by a week off the cycle then repeated for a second week. Patients were examined weekly until the lesions were eradicated. RESULTS: Clinical resolution of disease occurred in 18/20 cases. The mean time to resolution was 4.5 weeks, the mean number of cycles of treatment was two. Average follow up was 13 months with four cases of recurrent disease. These four cases were retreated with complete resolution in two cases. Epithelial toxicity occurred in 10/20 eyes and lid toxicity in two cases. There were no long term complications on discontinuing mitomycin C. CONCLUSIONS: Mitomycin C is effective in inducing regression of corneal conjunctival intraepithelial neoplasia. Complications are common but self-limiting. An optimal regimen is still to be established. PMID- 11886412 TI - The effect of water content on the 193 nm excimer laser ablation. AB - BACKGROUND: Water content of the corneal stroma may influence excimer laser ablation and may therefore affect residual refractive error following laser in situ keratomileusis. This study reports associations between water content of hydrogel materials and laser ablation depth. METHODS: Hydrated (n = 4) and dehydrated (n = 4) hydrogel buttons of 38%, 45%, 55% and 69% water content were ablated with the Nidek EC-5000 ArF 193 nm excimer laser, set to deliver a -6.00 DS curvature. Central curvature, optical quality and water content were measured before and after ablation. Hydrated buttons were rehydrated postablation and prior to measurement, to eliminate the effect of water removal during the procedure. The ablation depth per pulse was calculated. RESULTS: The average ablation rate for fully hydrated buttons was 0.51 +/- 0.17 microm. The ablation rate for hydrated materials (dry component ablation) reduced with increasing water content (P < 0.001). Dry hydrogel materials (0% water content) had an average ablation rate of 0.23 +/- 0.06 microm per pulse. CONCLUSIONS: For a constant laser energy output, lower water content materials ablated to a greater extent than higher water content materials. This model provides a simple way to assess the effect of water content and dehydration on myopic laser in situ keratomileusis. PMID- 11886413 TI - Factors affecting awareness and knowledge of glaucoma among patients presenting to an urban emergency department. AB - BACKGROUND: Until advanced, glaucoma is asymptomatic. For early diagnosis to occur, patients may need to be aware of it and seek assessment regularly. People who have risk factors for glaucoma may have a greater awareness of the disease. METHODS: Patients presenting to an urban hospital emergency department were surveyed with a brief questionnaire to assess their knowledge of glaucoma. Data was collected about their gender, age, family history of glaucoma and presence of systemic hypertension, diabetes, Raynaud's phenomenon, migraines and myopia. RESULTS: Women (Odds ratio 2.3; 95% CI 1.4-3.7; P < 0.01), people who were 40 years or older (Odds ratio 2.2; 95% CI 1.1-4.4; P < 0.05) and those who were aware of a family history of glaucoma (Odds ratio 15.7; CI 5.5-45.3; P < 0.01) knew significantly more about the disease than others. People with other risk factors did not demonstrate significantly greater knowledge despite 89% of all participants having had a previous eye examination. CONCLUSION: This information may be useful to predict which patients may know about glaucoma when they present for an eye examination and who should be targeted in public health campaigns. PMID- 11886414 TI - Indocyanine green angiography in the presence of subretinal or intraretinal haemorrhages: clinical and experimental investigations. AB - PURPOSE: The absorption and emission characteristics of indocyanine green are associated with better penetration through ocular pigments, including melanin and blood, in comparison with fluorescein. Therefore, it has been assumed that indocyanine green angiography (ICG-A) allows better delineation of fluorescent structures including choroidal neovascularization in the presence of haemorrhages. The degree and frequency of blockage by haemorrhages during ICG-A and fluorescein angiography (Fl-A) were compared and absorption characteristics by blood were experimentally determined. METHODS: Simultaneous confocal scanning laser ophthalmoscopy was performed in patients with intraretinal or sub-retinal haemorrhages associated with various retinal diseases including neovascular age related macular degeneration. Areas of blocked choroidal fluorescence were compared in Fl-A and ICG-A using a standardized classification system by two independent readers. Experimental absorption measurements were performed using blood-filled quartz cuvettes and laser light with 488 and 790 nm, respectively. RESULTS: Sixty eyes of 59 patients were analysed. Twelve eyes (20%) showed blockage in Fl-A only corresponding with funduscopically visible blood. In 35 eyes (58%) the extent of absorption was greater in Fl-A compared with ICG-A. An identical area of blockage in both Fl-A and ICG-A was noted in 13 eyes (22%). The coefficient of absorption was 18.4 mm(-1) for Fl-A (488 nm) and 5.4 mm(-1) for ICG-A (790 nm). CONCLUSIONS: In contrast to previous assumptions, the findings indicate that clinically intraretinal or subretinal haemorrhages are frequently associated with blockage not only in Fl-A but also in ICG-A. This is in accordance with the experimentally determined coefficient of absorption. Apparently, haemorrhages occurring in association with retinal and choroidal diseases commonly have a thickness sufficient enough to induce relevant absorption during ICG-A, and thus impair delineation of fluorescent structures in planes posterior to the haemorrhage. Therefore, the diagnostic value of ICG-A in presence of subretinal or intra-retinal bleedings is limited. PMID- 11886415 TI - Delivery of photocoagulation treatment for diabetic retinopathy at a large Australian ophthalmic hospital: comparisons with national clinical practice guidelines. AB - OBJECTIVE: To determine if the delivery of photocoagulation for diabetic retinopathy at a large Australian ophthalmic hospital conforms with Australian National Health and Medical Research Council clinical practice guidelines. METHODS: A retrospective medical record review was conducted of all patients who had initial laser treatment for diabetic retinopathy at the Royal Victorian Eye and Ear Hospital from January 1997 to December 1998. RESULTS: The study included 322 eyes from 203 patients. The mean age was 65.8 years (range 18-89 years) and the mean duration of diabetes was 14.7 years (range 1-40 years). Panretinal photocoagulation (PRP) alone was performed in 37 eyes over a mean number of 2.6 sessions. The median waiting time for these procedures was 14 days (range 0-146 days) and the median follow-up time was 7.1 weeks (range 1-25 weeks). Focal treatment was performed (without PRP) in 238 eyes and 55.5% of these cases required repeat focal treatment for persistent clinically significant macular oedema. Median waiting time for focal treatment was 20 days (range 0-302 days) and the median follow-up time after treatment was 12.1 weeks (range 1.7-42.0 weeks). Focal and PRP treatment was used in 47 eyes that had maculopathy concurrently with proliferative retinopathy. Focal treatment was applied before (or at the same session as) the PRP wherever possible. CONCLUSIONS: The study indicates that the application of photocoagulation and follow up for diabetic retinopathy at this tertiary referral institution conforms closely with Australian clinical practice guidelines. PMID- 11886416 TI - A study of the indications and changing trends of evisceration in north India. AB - AIM: To study the demographic pattern and indications for evisceration in north India and to evaluate the changing trends over the last decade. METHODS: In a retrospective hospital-based study, case records of all patients who underwent evisceration at Rajendra Prasad Centre for Ophthalmic Sciences from January 1990 to December 1999 were reviewed. The parameters evaluated were the age and sex distribution, the place of residence (urban/rural) and the indications for evisceration. The aetiology responsible for evisceration was determined on the basis of history, clinical examination and investigations as determined from previous records. RESULTS: One hundred and sixty-four patients had one eye eviscerated during the study period. The mean age of the patients was 51 +/- 13.84 years (range 6 months to 90 years). Panophthalmitis was the most common indication for evisceration (78.6%, n = 129), followed by irreparable globe injury (21.3%, n = 35). There was a significant decrease in the eviscerations performed due to pano-phthalmitis from 104 cases during the period 1990-1994, to 25 cases in the period 1995-1999. CONCLUSION: Panophthalmitis and severe ocular injury are the major indications of evisceration in north India. There has been a significant decrease in the number of eviscerations related to panophthalmitis over the last decade. PMID- 11886417 TI - The location of insulin receptors in bovine retina and isolated retinal cells. AB - PURPOSE: The binding of insulin to its cell-surface receptor is the sole means by which the hormone influences cellular activity. The location of insulin receptors in bovine retina and on isolated retinal cells was investigated to determine the specific cells sensitive to insulin. METHODS: Insulin receptors were located in frozen retinal sections prepared from enucleated bovine eyes, with polyclonal anti-insulin receptor antibodies using an immuno-peroxidase method. Isolated cells were obtained by enzymatic and physical dispersion of bovine retinal tissue. Insulin receptors on isolated cells were located by a monoclonal anti insulin receptor antibody using an immunogold silver staining technique. RESULTS: Insulin receptors demonstrated a widespread distribution throughout the bovine retina, being present in all retinal layers. A particular association with the plexiform layers and Muller cells was identified in the frozen sections. Consistent with these findings, insulin receptors were predominantly located on dendritic processes of isolated retinal neurones and on Muller cells. CONCLUSIONS: The widespread distribution of retinal insulin receptors in the bovine retina supports the hypothesis that insulin has a role in regulating retinal activity. Insulin receptors associated with plexiform regions suggests that insulin may influence neural activity, while receptors on Muller cells indicate that insulin may have a role in metabolic or functional mechanisms in bovine retina. PMID- 11886418 TI - Quantitative analysis of apoptosis in retinoblastoma. AB - PURPOSE: The rate of apoptosis in retinoblastoma (Rb) and the factors that may influence this rate, such as therapy prior to surgery, amount of necrosis in tumour tissue, differentiation and laterality of the tumour, were investigated. METHODS: Thirty-one specimens (25 enucleation, six exenteration) with Rb were studied. Prior to final surgery, three patients received systemic chemotherapy, one intravitreal chemotherapy, one transpupillary thermotherapy, one external beam radiotherapy and one high-dose oral methylprednisolone therapy. The apoptotic index (AI,%) was calculated by counting at least 1000 cells under light microscopy (x 100) using TUNEL (terminal deoxynu-cleotidyl transferase-mediated dUTP nick-end labelling) method. RESULTS: The mean AI was 2.75 plus minus 1.2. No statistically significant association was observed between rate of apoptosis and the presurgical treatment, extent of necrosis, tumour differentiation and laterality. CONCLUSION: Apoptosis is an important mechanism of cell death, and may be a limiting factor for tumour progression. In this study, the rate of apoptosis was not affected by any of the studied parameters. PMID- 11886419 TI - The cutaneomarginal eyelid graft. AB - PURPOSE: To assess the effectiveness of the cutaneomarginal graft. METHODS: The cutaneomarginal graft consists of eyelid margin tissue with anterior lamella skin. The tarsal plate is resected following harvesting of a wedge of eyelid tissue. The graft is used to repair eyelid defects following excision of neoplasms which spares the posterior lamella. The graft is principally used at the lateral most part of the lower eyelid following Mohs surgery. A case series of five patients are reviewed to assess the effectiveness of the technique. RESULTS: All patients achieved a satisfactory result following grafting and there were no donor site complications. CONCLUSIONS: The cutaneomarginal graft is an effective means of repair, although with limited indications. PMID- 11886421 TI - Septic superior ophthalmic vein thrombosis. AB - An illustrative case of septic superior ophthalmic vein thrombosis secondary to a staphylococcus orbital cellulitis is presented and correlated with autopsy findings. A literature review and discussion of the risks and benefits of anti coagulation in this setting is outlined. PMID- 11886422 TI - Hypersensitivity reaction to intravitreal clindamycin therapy. AB - A Caucasian woman presenting with recurrence of intra-ocular toxoplasmosis was given intravitreal clindamycin. She subsequently developed a hypersensitivity reaction in the form of a generalized erythematous rash. To the authors' knowledge, hypersensitivity reactions to an antibiotic given by the intra-vitreal route have not previously been reported. PMID- 11886420 TI - Optic nerve cysticercosis. AB - Cysticercosis of the optic nerve is an extremely rare entity and only seven cases have been reported in the world literature. A case of optic nerve cysticercosis in a 25-year-old woman is reported, along with a review of literature. The patient presented with two episodes of pain, diminution of vision and proptosis. Computed tomography and ultrasonography revealed an intraneural cyst with scolex in the retrobulbar portion of the optic nerve. A positive enzyme-linked immunosorbent assay test for cysticercosis further confirmed the diagnosis. Medical therapy in the form of oral albendazole and steroids resulted in complete resolution of the cyst, with few visual sequelae. PMID- 11886423 TI - Epithelial corneal oedema treated with honey. PMID- 11886424 TI - Iron: an essential but potentially harmful nutrient. PMID- 11886425 TI - Haemochromatosis mutations and ferritin in myocardial infarction: a case-control study. AB - BACKGROUND: Iron accumulation may contribute to coronary heart disease by catalysing free radical formation and promoting oxidation of low-density lipoprotein cholesterol. Epidemiological studies of iron status and coronary heart disease are conflicting. DESIGN: To test whether genetic haemochromatosis is associated with myocardial infarction, we determined the prevalence of three mutations in the HFE gene (Cys282Tyr, His63Asp and Ser65Cys) in a 2 : 1 case control study including 177 patients who survived an acute myocardial infarction and 89 controls. Genotypes were determined by PCR amplification of genomic DNA followed by restriction enzyme digestion. We also studied the relationship between plasma ferritin and myocardial infarction. RESULTS: The carrier frequencies of these three mutations were not statistically different among patients and controls (Cys282Tyr: 1.4 vs. 10.1%; His63Asp: 26.5 vs. 31.5%; Ser65Cys: 2.8 vs. 1.1%). Mean ferritin levels were elevated among patients (176 +/- 155 microg L(-1)) compared with controls (131 +/- 106 microg L(-1), P = 0.015). Subjects with plasma ferritin concentrations of 300 microg L(-1) or more had a 2.9-fold (95% CI: 1.2-7.3, P = 0.02) unadjusted risk for a myocardial infarction compared with those with normal levels. In a univariate analysis, ferritin was significantly associated with myocardial infarction. Upon multiple regression analysis adjusting for smoking, hypertension, diabetes, body-mass index and total cholesterol, significance was no longer present. CONCLUSIONS: No direct association was found between genetic haemochromatosis and myocardial infarction among Swiss whites. Raised ferritin levels among patients suggest a role of increased iron stores in myocardial infarction, but iron overload was not an independent risk factor for Swiss coronary heart disease patients. PMID- 11886426 TI - Ferric saccharate induces oxygen radical stress and endothelial dysfunction in vivo. AB - BACKGROUND: Intravenous iron supplementation is used widely in haemodialysis patients. However, nontransferrin-bound iron (NTBI), which increases after intravenous supplementation of ferric saccharate, has been suggested to act as a catalytic agent in oxygen radical formation in vitro and may thus contribute to endothelial impairment in vivo. MATERIALS AND METHODS: In 20 healthy volunteers the effect of 100 mg ferric saccharate infusion was investigated. Vascular ultrasound was used to assess endothelium-dependent vasodilatation at baseline, and 10 and 240 min after ferric saccharate infusion. Whole blood was collected to measure NTBI and in vivo radical formation was assessed by electron spin resonance. A time-control study was performed using saline infusion. RESULTS: Infusion of ferric saccharate induces a greater than fourfold increase in NTBI, as well as a transient, significant (P < 0.01) reduction of flow-mediated dilatation 10 min after infusion of ferric saccharate, when compared with saline. The generation of superoxide in whole blood increased significantly 10 and 240 min after infusion of ferric saccharate by, respectively, 70 and 53%. CONCLUSIONS: Iron infusion at a currently used therapeutic dose for intravenous iron supplementation leads to increased oxygen radical stress and acute endothelial dysfunction. PMID- 11886427 TI - Iron does not bind to the Apo-B protein of low-density lipoprotein. AB - BACKGROUND: Epidemiological and experimental evidence suggests that (nontransferrin-bound) iron plays an important role in atherogenesis by catalysing peroxidation of low-density lipoprotein (LDL). However, the mechanism of the interaction of iron and LDL is unclear. Iron has to be in the closest vicinity of LDL in order to catalyse the formation of the short-lived hydroxyl. In this study we investigated whether iron can bind to LDL in order to facilitate LDL peroxidation. METHODS: LDL and [(59)Fe]ferric citrate were incubated at 37 degrees C and pH 7.4 for 30 min. Unbound [(59)Fe]ferric citrate was separated from LDL using a Sephadex G25-M column. Activity of [59Fe]ferric citrate was measured in the collected fractions. A control experiment was performed using albumin instead of LDL. RESULTS AND CONCLUSION: No binding was observed between iron, as a low molecular weight Fe(III) complex, and LDL. As a control albumin was able to bind iron, it seems evident that interaction of iron with LDL will involve other iron complexes. PMID- 11886428 TI - Red blood cell antioxidant and iron status in alcoholic and nonalcoholic cirrhosis. AB - BACKGROUND: Iron overload has been reported in alcoholic liver cirrhosis but it remains to be established whether iron is involved in inducing oxidative damage to erythrocytes in alcoholic cirrhosis. The aim of this study was to assess oxidative damage and red cell indicators of antioxidant defences in alcoholics with mild-to-severe liver cirrhosis, taking into account the iron status. MATERIALS AND METHODS: Twenty-nine patients with alcoholic liver cirrhosis (AC) and 27 with nonalcoholic cirrhosis (NAC) were studied. Serum lipid peroxides (LPO) were assayed by a colourimetric method. Serum-free malonyldialdehyde (MDA) was assayed by selected ion monitoring in positive chemical ionization; serum 4 hydroxy-2(E)-nonenal (4-HNE) was determined by a colorimetric method. Reduced (GSH) and oxidized glutathione (GSSG), adenine and pyridine cofactors were assayed in whole blood extracts by HPLC. Hexose-monophosphate shunt (HMPS), glycolytic pathway (EMP) and antioxidant enzyme activities were determined by standard methods. Iron status was evaluated by standard clinical chemistry and by histological grading of liver iron. Nontransferrin-bound iron (NTBI) was measured in serum by HPLC. RESULTS: GSH progressively decreased with increasing severity of liver involvement in AC and NAC. MDA, 4-HNE and NTBI were significantly higher in AC serum. Stimulation of red cell HMPS and reducing potential, in terms of NADPH production, were more pronounced in AC. CONCLUSIONS: These results suggest that NTBI is more important than the decrease of antioxidant defences in inducing lipid peroxidation. NTBI may play a catalytic role in free radical reactions in the presence of cellular reductants such as NADPH. PMID- 11886430 TI - Nontransferrin-bound iron in the plasma of haemodialysis patients after intravenous iron saccharate infusion. AB - BACKGROUND: Many haemodialysis patients treated with recombinant human erythropoietin (r-HuEPO) receive intravenous iron supplementation on a regular basis. It has been shown previously that this may result in a transient "oversaturation" of transferrin. METHODS: Ten stable haemodialysis patients on r HuEPO treatment received 100 mg iron saccharate in 60 min, and 1 week later 100 mg in 6 min. Conventional iron metabolism parameters and nontransferrin-bond iron, detected with HPLC after addition of nitrilotriacetate and pretreatment with cobalt, were measured. Also, iron was measured in dialysate. RESULTS: Serum iron increased from 9.6 +/- 6.2 to 213.7 +/- 49.4 micromol L(-1) (P < 0.001) when iron was given in 60 min, and from 11.1 +/- 4.7 to 219.3 +/- 43.7 micromol L(-1) (P < 0.001) when iron was given in 6 min. Transferrin saturation increased from 0.22 +/- 0.18 to 4.75 +/- 1.35 in protocol 1 and 0.26 +/- 0.16 to 4.91 +/- 1.38 in protocol 2. Nontransferrin-bound iron increased from 0.74 +/- 0.69 to 3.79 +/- 1.41 micromol L(-1) in protocol 1, and from 0.90 +/- 0.92 to 2.90 +/- 0.96 micromol L(-1) in protocol 2. No significant iron concentrations were found in dialysate before or during the iron saccharate infusion. CONCLUSION: Nontransferrin-bound iron exists in plasma of dialysis patients after infusion of iron saccharate. There was no difference when 100 mg iron was given in 60 min or in 6 min. Before iron infusion, appreciable concentrations of nontransferrin bound iron could already be detected. The clinical significance is not clear, but the findings may be important since nontransferrin-bound iron can act as a catalytic agent in the formation of hydroxyl radicals, thus potentially inducing cell damage and atherosclerosis. PMID- 11886429 TI - Iron, hepatic stellate cells and fibrosis in chronic hepatitis C. AB - BACKGROUND/AIMS: In patients with chronic hepatitis C, hepatic iron concentration correlates with liver fibrosis. However, it is not clear whether this correlation merely reflects the presence of more active disease, or iron exacerbates chronic hepatitis C virus (HCV)-induced damage through activation of hepatic stellate cells and regeneration of hepatocytes. MATERIALS AND METHODS: We studied 72 HCV positive patients, staged according to the Ishak's score system. We measured hepatic iron concentration with spectrophotometry and evaluated the number of hepatic stellate cells (using monoclonal antibody against alpha smooth muscle actin) and proliferating hepatocytes (using monoclonal antibody against Ki67). Iron and ferritin serum levels were also determined. RESULTS: Hepatic iron concentration correlated statistically with ferritin serum level (r = 0.59, P < 0.001), with grading (r = 0.47, P < 0.001) and staging (r = 0.51, P < 0.001) scores for chronic hepatitis in the whole group of patients. Hepatic iron concentration correlated positively with stellate cell number (r = 0.55, P = 0.004) and Ki67-positive hepatocyte number (r = 0.36, P = 0.08) in patients with chronic hepatitis C and low grading score (< 3). CONCLUSIONS: In patients with chronic hepatitis C and low grading score, hepatic iron could play a role in the activation of hepatic stellate cells and in the progression of fibrosis. PMID- 11886431 TI - Labile iron in parenteral iron formulations and its potential for generating plasma nontransferrin-bound iron in dialysis patients. AB - BACKGROUND: Labile plasma iron (LPI) associated with iron supplementation has been implicated in complications found in dialysis patients. As LPI can potentially catalyse oxygen radical generation, we determined the presence of labile iron in the parenteral preparations and the frequency of occurrence of LPI in dialysis patients. DESIGN: The capacity to donate iron to apotransferrin (apo ) or to the chelator desferrioxamine (DFO) was measured with fluorescein-Tf (Fl Tf) and Fl-DFO, respectively. Those probes undergo quenching upon binding to iron. Iron-catalysed generation of oxidant species was determined with dihydrorhodamine. Plasma nontransferrin-bound iron (NTBI), here termed LPI, was determined by mobilization of iron from low-affinity binding sites with oxalate, followed by its quantification with Fl-Tf in the presence of Ga(III). RESULTS: Normal individuals and most (80%) dialysis patients, analysed at least 1 week after iron supplementation showed no detectable (<0.2 microm) LPI. However, approximately 20% of the patients (n = 71) showed significant LPI levels (>0.2 microm), in some cases weeks after iron administration. LPI levels correlated best (r2 = 0.9) with Tf saturation. The iron preparations contained 2-6% low molecular weight and redox-active iron, most of which is chelated by Tf. CONCLUSIONS: Parenteral iron formulations contain a small but significant fraction of redox-active iron, most of which is scavenged by apo-Tf within <1 h. Therefore, oxidant stress associated with iron infusion is likely to be transient. The bulk of the polymeric iron is apparently inaccessible to apo-Tf. Although LPI might return to normal within 2 h of intravenous iron infusion, the long-term persistence of low-level LPI in up to 20% of end stage renal disease (ESRD) patients indicates that complete clearance of the intravenous iron may be more protracted than originally estimated. PMID- 11886432 TI - Nature of nontransferrin-bound iron. AB - Nontransferrin-bound iron is now recognized to exist by many workers. It is present in the serum of patients suffering from a wide range of disease states and may be induced by certain therapeutic treatments. The chemical nature of this iron pool is unknown but almost certainly it is a multicomponent pool including a considerable proportion of protein-bound iron. Methods are required to separate and quantify these different components. The biological properties of the individual isoforms need to be established; it is possible that some forms are relatively nontoxic, while others are highly toxic. This paper reviews what is known about the nature of nontransferrin-bound iron and describes the methods currently available to quantify this important serum component. PMID- 11886433 TI - Oxidative status and malondialdehyde in beta-thalassaemia patients. AB - BACKGROUND: In beta-thalassaemia syndromes, decreased or impaired biosynthesis of beta-globin leads to accumulation of unpaired alpha-globin chains. Moreover, the iron overload in beta-thalassaemia patients generates oxygen-free radicals and peroxidative tissue injury. The aim of this study was to detect and correlate iron overload parameters with the oxidative stress and the antioxidant capability in beta-thalassaemia patients. DESIGN: Serum iron, transferrin saturation, serum ferritin, nontransferrin-bound iron (NTBI), levels of serum free and total (free + bound) malondialdehyde (MDA) and total peroxyl radical-trapping antioxidant parameter (TRAP) were evaluated in 21 regularly transfused beta-thalassaemia major (TM) patients, 13 untransfused beta-thalassaemia intermedia (TI) patients and 17 healthy controls. Blood from the TM patients was drawn 48 h after the last desferoxamine (20-40 mg kg(-1)) infusion and just before transfusion. RESULTS: Free and total MDA and NTBI levels were higher in the TM patients than in the TI. In the TM patients the free MDA levels correlated positively with serum iron (r = +0.3, P = 0.0006), whereas the total MDA correlated positively with NTBI (r = +0.45, P = 0.037). However, a negative correlation was observed between TRAP and NTBI (r = -0.4, P = 0.0006). In the TI patients there was no significant correlation between free or total MDA and TRAP or NTBI. CONCLUSIONS: Our results confirm the peroxidative status generated by iron overload in thalassaemia patients and highlight the rapid formation of marked amounts of free MDA despite the chelation therapy in TM patients. PMID- 11886435 TI - Iron and immunity: a double-edged sword. AB - Iron is a crucial element for many central metabolic pathways of the body. Lack of iron leads to growth arrest and anaemia while increased accumulation of this metal, as it occurs in highly frequent inherited diseases such as hereditary haemochromatosis and thalassaemia, is associated with toxic radical formation and progressive tissue damage. As shown by several groups, iron also modulates immune effector mechanisms, such as cytokine activities (IFN-gamma effector pathways towards macrophages), nitric oxide (NO) formation or immune cell proliferation, and thus host immune surveillance. Therefore, gaining control over iron homeostasis is one of the central battlefields in deciding the fate of an infection with intracellular pathogens or a malignant disease. Thus, the reticulo endothelial system has evoked sophisticated strategies to control iron metabolism in general and especially the handling of the metal within immune cells. PMID- 11886434 TI - Transferrin toxin but not transferrin receptor immunotoxin is influenced by free transferrin and iron saturation. AB - BACKGROUND: Cytotoxic agents can be targeted successfully to cancer cells. The efficacy of such novel and potent anticancer strategies may be influenced by variables of iron metabolism. METHODS: The in vitro cytotoxicity against glioma cells of transferrin (Tf)-based targeted toxins was compared with that of alpha transferrin receptor (TfR)-immunotoxin. RESULTS: Of four Tf-based targeted toxins, Tf-gelonin, Tf-pokeweed antiviral protein, Tf-momordin and Tf-saporin, inhibitory concentration 50% values against glioma-derived cell lines HS683 and U251, ranged from [4.8 +/- 1.5] x 10(-10) m for Tf-saporin to [26.9 +/- 15.3] x 10(-10) m for Tf-gelonin in [(3)H]-leucine incorporation assays. Tf-saporin and alpha-TfR-saporin-immunotoxin had similar efficacy, even in the more quantitative clonogenic assay (4-5 log kill with 1 x 10(-9) m) using the myeloma cell line RPMI 8226 and glioma cell line U251. However, on RPMI 8226, the efficacy of Tf saporin 1 x 10(-9) m was reduced by 90% in the presence of 150 microg mL(-1)(=20% of normal plasma value) competing diferric transferrin, whereas the efficacy of the corresponding immunotoxin was affected only marginally. In addition, the efficacy of Tf-based conjugates will depend on their iron saturation state. Iron desaturation of Tf-saporin was demonstrated by [(59)Fe]-labelling, subsequent CM Sepharose chromatography and SDS-PAGE. Desaturation led to virtually complete loss of affinity for the transferrin receptor, as determined by flow cytometry, which could be largely restored upon resaturation. CONCLUSION: Transferrin-based toxin conjugates are strongly influenced by the presence of free transferrin and the iron saturation state. The corresponding alpha-transferrin receptor immunotoxin does not show these disadvantages, has similar efficacy and should be preferred for further experiments. PMID- 11886436 TI - Regulation of ferritin: a specific role for interferon-alpha (IFN-alpha)? The acute phase response in patients treated with IFN-alpha-2b. AB - BACKGROUND: Adult onset of Still's disease is characterized by very high serum ferritin levels, in disproportion with other acute phase proteins (APPs). Because interferon-alpha (IFN-alpha) was observed to cause hyperferritinaemia in three healthy people without increase of other APPs, we hypothesized that IFN-alpha stimulates specifically the synthesis of ferritin. To test this hypothesis, we studied ferritin and other APP levels in patients treated with IFN-alpha. PATIENTS AND METHODS: Fifteen patients treated with IFN-alpha-2b 3-5 times a week, as adjuvant treatment after excision of a high-risk melanoma, were compared with six patients without adjuvant treatment (controls). Serum levels of C reactive protein (CRP) and secretory phospholipase A2 (sPLA2) were measured using ELISA. Levels of ferritin, alpha1-acid glycoprotein (AAG) and albumin were determined by nephelometry. RESULTS: CRP was decreased significantly after 4 weeks (P < 0.01) in the patients treated with IFN-alpha compared with the nontreated patients, after 6 months of treatment it was still decreased although not significantly. Ferritin increased significantly in the IFN-alpha-treated patients: 187% of pretreatment value after 4 weeks and 217% after 6 months (P < 0.01), while ferritin levels decreased in the nontreated patients. AAG increased significantly in IFN-alpha-treated patients (107, 114%) compared with the control patients (91, 76%) but differences were less compared with CRP and ferritin. sPLA2 had a variable course, while albumin remained constant within the normal range in both patient groups. CONCLUSIONS: IFN-alpha induced a significant increase in ferritin, with a significant decrease in CRP, little increase in AAG, varying response of sPLA2 and no change in albumin. This finding suggests a specific role for IFN-alpha in the synthesis or secretion of ferritin. This mechanism may also be involved in the marked hyperferritinaemia in adult onset of Still's disease. PMID- 11886437 TI - Iron chelation and hydroxyl radical scavenging reduce the inflammatory response of endothelial cells after infection with Chlamydia pneumoniae or influenza A. AB - BACKGROUND: Chronic low-grade inflammation is associated with increased risk of vascular diseases. The source of inflammation is unknown but may well be chronic and/or repetitive infections with microorganisms. Direct infection of endothelial cells (ECs) may also be a starting point for atherogenesis by initiating endothelial procoagulant activity, increased monocyte adherence and increased cytokine production. We hypothesized that iron-mediated intracellular hydroxyl radical formation after infection is a key event in triggering the production of interleukin-6 (IL-6) by ECs in vitro. METHODS: Cultured ECs were incubated with Fe(II) and Fe(III) or infected with Chlamydia pneumoniae or influenza A/H1N1/Taiwan/1/81 for 48 and 24 h, respectively. To determine the role of iron and reactive oxygen species, cells were coincubated with the H2O2 scavenger N acetyl-l-cysteine, with the iron chelator deferoxamine (DFO) or with the intracellular hydroxyl radical scavenger dimethylthiourea (DMTU). After the incubation periods, supernatants were harvested for IL-6 determination. RESULTS: Incubating ECs with Fe(II) and Fe(III) resulted in increased IL-6 production. Similarly, infection with C. pneumoniae and influenza A also induced an IL-6 response. Coincubating ECs with DFO or DMTU blocked this response. Nuclear factor kappaB activity was increased after infection and blocked by coincubation with DFO or DMTU. CONCLUSION: Cultured ECs respond to infection and iron incubation with increased production of IL-6. Iron, the generation of intracellular hydroxyl radical and NF-kappaB activity are essential in cellular activation, suggesting that reactive oxygen species generated in the Haber-Weiss reaction are essential in invoking an immunological response to infection by ECs. PMID- 11886438 TI - Human immunodeficiency virus type 1 replication inhibition by the bidentate iron chelators CP502 and CP511 is caused by proliferation inhibition and the onset of apoptosis. AB - BACKGROUND: The iron chelators deferoxamine (DF) and deferiprone (CP20) have been shown to inhibit human immunodeficiency virus type 1 (HIV-1) replication in human peripheral blood lymphocytes (PBL). The orally active bidentate chelators CP502 and CP511, which also belong to the 3-hydroxypyridin-4-one family, but with higher affinities for iron than CP20, were monitored for their antiviral properties by checking for p24 antigen production and nuclear factor (NF)-kappaB activation, and their ability to induce apoptosis. MATERIALS AND METHODS: Human PBLs were isolated from HIV-1 seronegative donors and subsequently infected with HIV-1(Ba-L) for 2 h. After 5 days' incubation, HIV-1 replication was monitored by p24 antigen production. Cellular proliferation as well as caspase-3 activity were monitored in uninfected cells after a period of 5 days and after 1 day infection, respectively. NF-kappaB activity was also monitored by electromobility shift assays (EMSA) performed on nuclear extracts of Jurkat cells treated with the different chelators for 4 h. RESULTS: CP502 and CP511 decrease HIV-1 replication by decreasing cellular proliferation in a similar manner to DF and CP20. CP511 seemed to be more potent than either CP502 or CP20. Due to the reduction in cellular proliferation, there was an increase in caspase-3 activity after 24 h incubation. NF-kappaB activity was not affected by any of the chelators. CONCLUSIONS: Iron chelators with high affinities for iron, which are under development for the treatment of iron overload, could contribute to the reduction of HIV-1 replication in infected patients by cellular proliferation inhibition rather than by a direct antiviral action. PMID- 11886439 TI - Somatosensory cortical neuronal population activity across states of anaesthesia. AB - Experiments were carried out to learn about changes in sensory cortical processing associated with different levels of anaesthesia. Traditionally this question has been addressed by studying single neurons. Because state changes are likely to influence the relationships between neurons, the present experiments were undertaken to investigate the spatial and temporal firing patterns distributed across cortex. Using 5 x 5 or 10 x 10 microelectrode arrays, spontaneous and stimulus-evoked activity of multineuron clusters was recorded from rat somatosensory 'barrel' cortex (the whisker representation) during a light surgical stage of urethane anaesthesia, and after two supplemental doses of urethane which led to intermediate and deep levels of anaesthesia. At all depths of anaesthesia, spontaneously occurring action potentials at a single electrode tended to be clustered into 'bursts.' With increasing anaesthetic depth, bursts became more prominent and rhythmic, and increasingly synchronized between cortical barrel-columns. Burst frequency decreased and fewer spikes occurred outside bursts, leading to a decrease in the overall spontaneous firing rate. The cortical territory engaged by individual whiskers contracted with increasing depth of anaesthesia, leading to the spatial segregation of whisker representations. At all stages of anaesthesia, whisker stimulation produced the maximal cortical response when delivered close to burst onset. These observations show that ongoing spontaneous activity modulates sensory response properties and makes peripheral tactile information accessible to a cortical territory whose size is determined by the phase of burst cycle. The possible significance of the cyclic cortical responsiveness encountered during urethane anaesthesia to cortical processing in awake rats is considered. PMID- 11886440 TI - GDNF and NGF released by synthetic guidance channels support sciatic nerve regeneration across a long gap. AB - The present work was performed to determine the ability of neurotrophic factors to allow axonal regeneration across a 15-mm-long gap in the rat sciatic nerve. Synthetic nerve guidance channels slowly releasing NGF and GDNF were fabricated and sutured to the cut ends of the nerve to bridge the gap. After 7 weeks, nerve cables had formed in nine out of ten channels in both the NGF and GDNF groups, while no neuronal cables were present in the control group. The average number of myelinated axons at the midpoint of the regenerated nerves was significantly greater in the presence of GDNF than NGF (4942 +/-1627 vs. 1199 +/-431, P < or = 0.04). A significantly greater number of neuronal cells in the GDNF group, when compared to the NGF group, retrogradely transported FluoroGold injected distal to the injury site before explantation. The total number of labelled motoneurons observed in the ventral horn of the spinal cord was 98.1 +/-23.4 vs. 20.0 +/-8.5 (P < or = 0.001) in the presence of GDNF and NGF, respectively. In the dorsal root ganglia, 22.7% +/- 4.9% vs. 3.2% +/-1.9% (P +/-0.005) of sensory neurons were labelled retrogradely in the GDNF and NGF treatment groups, respectively. The present study demonstrates that, sustained delivery of GDNF and NGF to the injury site, by synthetic nerve guidance channels, allows regeneration of both sensory and motor axons over long gaps; GDNF leads to better overall regeneration in the sciatic nerve. PMID- 11886442 TI - A dose-dependent facilitation and inhibition of peripheral nerve regeneration by brain-derived neurotrophic factor. AB - The time-dependent decline in the ability of motoneurons to regenerate their axons after axotomy is one of the principle contributing factors to poor functional recovery after peripheral nerve injury. A decline in neurotrophic support may be partially responsible for this effect. The up-regulation of BDNF after injury, both in denervated Schwann cells and in axotomized motoneurons, suggests its importance in motor axonal regeneration. In adult female Sprague Dawley rats, we counted the number of freshly injured or chronically axotomized tibial motoneurons that had regenerated their axons 1 month after surgical suture to a freshly denervated common peroneal distal nerve stump. Motor axonal regeneration was evaluated by applying fluorescent retrograde neurotracers to the common peroneal nerve 20 mm distal to the injury site and counting the number of fluorescently labelled motoneurons in the T11-L1 region of the spinal cord. We report that low doses of BDNF (0.5-2 microg/day for 28 days) had no detectable effect on axonal regeneration after immediate nerve repair, but promoted axonal regeneration of motoneurons whose regenerative capacity was reduced by chronic axotomy 2 months prior to nerve resuture, completely reversing the negative effects of delayed nerve repair. In contrast, high doses of BDNF (12-20 microg/day for 28 days) significantly inhibited motor axonal regeneration, after both immediate nerve repair and nerve repair after chronic axotomy. The inhibitory actions of high dose BDNF could be reversed by functional blockade of p75 receptors, thus implicating these receptors as mediators of the inhibitory effects of high dose exogenous BDNF. PMID- 11886441 TI - Spinal cord injury induces expression of RGS7 in microglia/macrophages in rats. AB - RGS proteins regulate G protein-mediated signalling pathways through direct interaction with the Galpha subunits and facilitation of GTP hydrolysis. An RGS subfamily consisting of RGS 6, 7, 9, and 11 also interacts with the G protein beta subunit Gbeta5 via a characteristic Ggamma-like domain. Thus far, these complexes were found only in neurons, with RGS7 being the most widely distributed in the brain. Here we confirm the expression of RGS7 in spinal neurons and show as a novel finding that following an experimental spinal cord injury in rats, expression of RGS7 is induced in a subpopulation of other cells. Immunofluorescent confocal microscopy using a series of cell specific antibodies identified these RGS7 positive cells as activated microglia and/or invading peripheral macrophages. To rule out interference from the adjacent neurons and confirm the presence of RGS7-Gbeta5 complex in inflammatory cells, we performed immunocytochemistry, RT-PCR, Western blot, and immunoprecipitation using microglial (BV2) and peripheral macrophage (RAW) cell lines. Expression of RGS7 mRNA and protein are nearly undetectable in non-stimulated BV2 and RAW cells, but remarkably increased after stimulation with LPS or TNF-alpha In addition, RGS7 positive cells were also found in the perinodular rim in the rat spleen. Our findings show that RGS7-Gbeta5 complex is expressed in immunocompetent cells such as resident microglia and peripheral macrophages following spinal cord injury. This expression might contribute to the post-traumatic inflammatory responses in the central nervous system. PMID- 11886443 TI - Differential post-transcriptional regulation of p21WAF1/Cip1 levels in the developing nervous system following gamma-irradiation. AB - Radiation-induced death in the developing brain is p53-dependent. However, genetic studies indicate that the signalling pathways that couple irradiation to p53 expression can vary between different developing neural populations [Herzog et al. (1998) Science, 280, 1089-1091]. Here we establish that signalling downstream of p53 also exhibits brain region-specific differences that are associated with the relative vulnerability of some cell populations to radiation induced killing in the mouse. Following gamma-irradiation, p53 and p21WAF1/cip1, but not Bax, protein levels increased in the developing cerebellum. In contrast, neither p21WAF1/cip1 nor Bax protein levels were elevated in the retina following irradiation, despite increased p53 expression. In the retina, p53 expression was associated with cells destined to die, whereas in the cerebellum, p53 was expressed in both radiation-sensitive and radiation-resistant neuroblasts of the external granule cell layer. Although p21WAF1/cip1 mRNA was expressed in all p53 positive neuroblasts after irradiation, p21WAF1/cip1 protein was only detected in radiation-resistant neuroblasts of the cerebellum. Thus, p21WAF1/cip1 was subject to post-transcriptional regulation with p21WAF1/cip1 protein only accumulating in cells destined to survive irradiation. Nevertheless, p21WAF1/cip1 function was not essential for radiation resistance, as postmitotic neuroblasts in the external granule cell layer were spared in p21WAF1/cip1 knockout mice. PMID- 11886444 TI - Dose-dependent rescue of axotomized rat retinal ganglion cells by adenovirus mediated expression of glial cell-line derived neurotrophic factor in vivo. AB - Adult rat retinal ganglion cells undergo degeneration after optic nerve transection. Repeated intraocular injection of glial cell-line derived neurotrophic factor (GDNF) has been shown to be efficient in enhancing retinal ganglion cell survival following optic nerve axotomy. In the present study we evaluated the potential survival-promoting effect of adenovirally administered GDNF on axotomized retinal ganglion cells. A single intravitreal injection [7 x 107 plaque-forming units (pfu) or 7 x 108 pfu] of an adenoviral vector expressing the rat GDNF gene from a cytomegalovirus promoter enhanced retinal ganglion cell survival 14 days after axotomy by 67 and 125%, respectively, when compared to control animals. Intraocular administration of the vector rescued 12.6 and 23%, respectively, of the retinal ganglion cells which would otherwise have died after axotomy. An increase in retinal GDNF protein and specific virally transduced GDNF mRNA expression was detected following intraocular vector application. Our data support previous findings showing that adenoviral delivery of neurotrophic factors to the vitreous body is a feasible approach for the prevention of axotomy induced retinal ganglion cell death in vivo and may constitute a relevant strategy for future treatment in traumatic brain injury and ensuing neurodegeneration. PMID- 11886445 TI - Hypothalamic bHLH transcription factors are novel candidates in the regulation of energy balance. AB - The basic helix-loop-helix transcription factors, neurological basic-helix-loop helix-2 (Nhlh-2), neurogenic differentiation-1 (NeuroD-1) and single minded-1 (Sim-1) could have roles in energy balance regulation, although supporting evidence is inconclusive. This study in mice provides further evidence that Nhlh 2 and NeuroD-1 are involved in energy balance regulation. In situ hybridization was used to study the expression of the genes in relation to physiological status and genetic background within hypothalamic nuclei that are involved in energy balance regulation. These studies show reduced expression of Nhlh-2 mRNA in the arcuate (ARC) nucleus and NeuroD-1 mRNA in the paraventricular (PVN) nucleus in obese ob/ob and 24 h food-deprived mice relative to respective controls, suggesting regulation by leptin. Interestingly, Nhlh-2 mRNA expression is reduced in obese db/db mice, whereas NeuroD-1 remains unchanged, suggesting different mechanisms of regulation by leptin of these two genes. To study the role of leptin in the regulation of these genes, leptin was injected intraperitoneally in obese ob/ob mice and mRNA expression evaluated after 1 h or 4 h, or after twice daily injection for 7 days. None of these regimes restored Nhlh-2 or NeuroD-1 to wild-type mRNA levels. These latter data suggest either that the regulation of the Nhlh-2 and NeuroD-1 genes by leptin is indirect or that the apparent leptin insensitivity of the gene expression reflects a developmental deficit that is a consequence of the phenotype of the obese ob/ob mice. The relationship between Nhlh-2 and candidate energy balance-related genes was studied by dual in situ hybridization. Nhlh-2 mRNA was coexpressed in a subpopulation (30%) of ARC neurons expressing pro-opiomelanocortin (POMC) mRNA, suggesting a potential functional relationship. PMID- 11886446 TI - In vivo activation and nuclear translocation of phosphorylated glycogen synthase kinase-3beta in neuronal apoptosis: links to tau phosphorylation. AB - The roles of glycogen synthase kinase-3beta (GSK-3beta) and tau phosphorylation were examined in seven-day-old rats injected with the NMDA receptor antagonist (MK801) that is known to induce neuronal apoptosis. Immunoblot and immunohistochemical analysis of brain samples demonstrated a site-specific increase in tau phosphorylation associated with the relocalization of the protein to the nuclear/perinuclear region of apoptotic neurons. In addition, a tau 32-kDa fragment was detected, suggesting that tau was a target of intracellular proteolysis in MK801-treated brains. The proteolytically modified form of tau has reduced ability to bind to microtubules. GSK-3beta kinase assay and immunoblottings of active (tyrosine-216) and inactive (serine-9) forms of GSK 3beta revealed a rapid and transient increase in the kinase activity. Lithium chloride, a GSK-3beta inhibitor, prevented tau phosphorylation suggesting that tau phosphorylation is mediated by the activation of GSK-3beta. Confocal microscopy using double labelling of tau and GSK-3beta revealed that the activation of GSK-3beta in neurons was associated with early (2 h) nuclear translocation of tyrosine-216 GSK-3beta. The execution phase of neuronal apoptosis was accompanied by a selective phosphorylation of serine-9 and dephosphorylation of tyrosine-216 GSK-3beta. These findings demonstrate that in vivo, GSK-3beta kinase activation and nuclear translocation are early stress signals of neuronal apoptosis. PMID- 11886447 TI - Integration of calcium signals by calmodulin in rat sensory neurons. AB - We have used the fluorescently labelled calmodulin TA-CaM to follow calmodulin activation during depolarization of adult rat sensory neurons. Calcium concentration was measured simultaneously using the low affinity indicator Oregon Green BAPTA 5N. TA-CaM fluorescence increased during a 200-ms depolarization but then continued to increase during the subsequent 500 ms, even though total cell calcium was falling at this time. In the next few seconds TA-CaM fluorescence fell, but to a new elevated level that was then maintained for several tens of seconds. During a train of depolarizations that evoked a series of largely independent calcium changes TA-CaM fluorescence was in contrast raised for the duration of the train and for many tens of seconds afterwards. The presence of a peptide corresponding to the calmodulin binding domain of myosin light chain kinase significantly increased the depolarization-induced TA-CaM fluorescence increase and slowed the subsequent fall of fluorescence. We interpret the slow recovery component of the TA-CaM signal as reflecting the slow dissociation of calcium--calmodulin--calmodulin binding protein complexes. Our results show that after brief electrical activity calmodulin's interaction with calmodulin binding proteins persists for approximately one minute. PMID- 11886448 TI - Mechanisms controlling bursting activity induced by disinhibition in spinal cord networks. AB - Disinhibition reliably induces regular synchronous bursting in networks of spinal interneurons in culture as well as in the intact spinal cord. We have combined extracellular multisite recording using multielectrode arrays with whole cell recordings to investigate the mechanisms involved in bursting in organotypic and dissociated cultures from the spinal cords of embryonic rats. Network bursts induced depolarization and spikes in single neurons, which were mediated by recurrent excitation through glutamatergic synaptic transmission. When such transmission was blocked, bursting ceased. However, tonic spiking persisted in some of the neurons. In such neurons intrinsic spiking was suppressed following the bursts and reappeared in the intervals after several seconds. The suppression of intrinsic spiking could be reproduced when, in the absence of fast synaptic transmission, bursts were mimicked by the injection of current pulses. Intrinsic spiking was also suppressed by a slight hyperpolarization. An afterhyperpolarization following the bursts was found in roughly half of the neurons. These afterhyperpolarizations were combined with a decrease in excitability. No evidence for the involvement of synaptic depletion or receptor desensitization in bursting was found, because neither the rate nor the size of spontaneous excitatory postsynaptic currents were decreased following the bursts. Extracellular stimuli paced bursts at low frequencies, but failed to induce bursts when applied too soon after the last burst. Altogether these results suggest that bursting in spinal cultures is mainly based on intrinsic spiking in some neurons, recurrent excitation of the network and auto-regulation of neuronal excitability. PMID- 11886450 TI - Cross-modal neuroplasticity in neonatally enucleated hamsters: structure, electrophysiology and behaviour. AB - Potential auditory compensation in neonatally bilaterally enucleated Syrian hamsters was explored anatomically, electrophysiologically and behaviourally. Gross morphology of the visual cortex appeared normal and no obvious cytoarchitectural malformation was discerned. However, enucleation induced a significant increase in the spontaneous firing rate of visual cortex cells. Further, auditory stimuli elicited field potentials and single unit responses in the visual cortex of enucleated, but not normal, animals. About 63% of the cells isolated in the visual cortex of 16 enucleated hamsters responded to at least one type of auditory stimulus. Most of the responses were less vigorous and less time locked than those of auditory cortex cells, and thresholds were typically higher. Projection tracing with WGA-HRP disclosed reciprocal connections between the visual cortex and the dorsal lateral geniculate nucleus in both intact and enucleated animals. However, in the enucleated animals retrogradely labelled cells were also found in the inferior colliculus, the major midbrain auditory nucleus. Behaviourally determined auditory sensitivity across the hearing range did not differ between enucleated and intact hamsters. Minimum audible angle, as determined by a conditioned suppression task, ranged from around 17 to 22 degrees, with no significant difference between normal and enucleated animals. The two groups also did not differ with regard to the direction of their unconditioned head orientating response to intermittent noise. However, the enucleated animals showed a more vigorous response and were slower to habituate to the noise. These results show that bilateral enucleation of newborn hamsters results in auditory activation of visual targets, in addition to the typical activation of the intact auditory pathway. Behaviourally it appears that enucleated hamsters, compared with their normal littermates, are slower to habituate in their response to an unexpected source of sound. PMID- 11886449 TI - Different respiratory control systems are affected in homozygous and heterozygous kreisler mutant mice. AB - During embryonic development, restricted expression of the regulatory genes Krox20 and kreisler are involved in segmentation and antero-posterior patterning of the hindbrain neural tube. The analysis of transgenic mice in which specific rhombomeres (r) are eliminated points to an important role of segmentation in the generation of neuronal networks controlling vital rhythmic behaviours such as respiration. Thus, elimination of r3 and r5 in Krox20-/- mice suppresses a pontine antiapneic system (Jacquin et al., 1996). We now compare Krox20-/- to kreisler heterozygous (+/kr) and homozygous (kr/kr) mutant neonates. In +/kr mutant mice, we describe hyperactivity of the antiapneic system: analysis of rhythm generation in vitro revealed a pontine modification in keeping with abnormal cell specifications previously reported in r3 (Manzanares et al., 1999b). In kr/kr mice, elimination of r5 abolished all +/kr respiratory traits, suggesting that +/kr hyperactivity of the antiapneic system is mediated through r5-derived territories. Furthermore, collateral chemosensory pathways that normally mediate delayed responses to hypoxia and hyperoxia were not functional in kr/kr mice. We conclude that the pontine antiapneic system originates from r3r4, but not r5. A different rhythm-promoting system originates in r5 and kreisler controls the development of antiapneic and chemosensory signal transmission at this level. PMID- 11886451 TI - Differential effects of typical and atypical antipsychotic drugs on striosome and matrix compartments of the striatum. AB - Administration of typical antipsychotic drugs (APDs) is often accompanied by extrapyramidal side-effects (EPS). Treatment with atypical APDs has a lower incidence of motor side-effects and atypical APDs are superior to typical APDs in treating the negative symptoms of schizophrenia. Although typical APDs strongly induce the immediate-early gene c-fos in the striatum while atypical APDs do so only weakly, it is possible that the effects of atypical APDs are more pronounced within certain regions of the striatum. The striatum contains two histochemically defined compartments, the striosome (patch) and the matrix. These compartments have been well characterized anatomically but their functional attributes are unclear. We therefore examined the effects of typical and atypical APDs on Fos expression in the striosome and matrix of the rat. Typical and atypical APDs were distinguished by the pattern of striatal compartmental activation they induced: the striosome : matrix ratio of Fos-li neurons was greater in rats treated with atypical APDs. Pretreating animals with selective antagonists of receptors that atypical APDs target with high affinity did not increase the striosome : matrix Fos ratio of typical APD-treated rats and thus did not mimic the ratio seen in response to atypical APDs. However, pretreatment with the atypical APD clozapine did recapitulate the characteristic compartmental Fos pattern seen in response to typical APDs. These data suggest that some characteristics of atypical APDs, such as the lower EPS liability and greater reduction of negative symptoms, may be linked to the coordinate regulation of the striatal striosome and matrix. PMID- 11886452 TI - Decreased BDNF signalling in transgenic mice reduces epileptogenesis. AB - Brain derived neurotrophic factor (BDNF) has been suggested to be involved in epileptogenesis. Both pro- and antiepileptogenic effects have been reported, but the exact physiological role is still unclear. Here, we investigated the role of endogenous BDNF in epileptogenesis by using transgenic mice overexpressing truncated trkB, a dominant negative receptor of BDNF. After induction of status epilepticus (SE) by kainic acid, the development of spontaneous seizures was monitored by video-EEG system. Hilar cell loss, and the number of neuropeptide Y immunoreactive cells were studied as markers of cellular damage, and mossy fibre sprouting was investigated as a plasticity marker. Our results show that transgenic mice had significantly less frequent interictal spiking than wild-type mice, and the frequency of spontaneous seizures was lower. Furthermore, compared to wild-type animals, transgenic mice had less severe seizures with later onset and mortality was lower. In contrast, no differences between genotypes were observed in any of the cellular or plasticity markers. Our results suggest that transgenic mice with decreased BDNF signalling have reduced epileptogenesis. PMID- 11886454 TI - Neuronal responses of the rat amygdala during extinction and reassociation learning in elementary and configural associative tasks. AB - To investigate functional heterogeneity within the amygdala in appetitive conditioned instrumental behaviours, neuronal activity was recorded from the amygdala of behaving rats during learning and discrimination of conditioned sensory stimuli associated with or without reinforcement [sucrose solution, intracranial self-stimulation (ICSS)]. Sensory stimuli included auditory (tone), visual (light) and configural (simultaneous presentation of tone and light) stimuli. The rat was trained to lick a spout protruded close to its mouth just after a conditioned sensory stimulus to obtain a reward. Of the 609 neurons recorded from the amygdala and amygdalostriatal transition area, 154 responded to one or more sensory stimuli. The 62 amygdalar neurons responded strongly to certain conditioned sensory stimuli associated with rewards. Of these 62 neurons, 45 were tested with the extinction trials. Responses of 31 neurons to conditioned stimuli were finally extinguished, and those of the remaining 14 were not extinguished. Furthermore, responses of 26 of these 31 neurons resumed in the relearning trials (plastic neurons), suggesting that these sensory responses were associative rather than just responses to physical properties of the stimuli. These plastic neurons were located mainly in the basolateral nucleus of the amygdala, and responses of the plastic neurons were correlated with behavioural responses. These results suggest that the basolateral nucleus is crucial in associative learning between sensory information and affective significance for behavioural outputs in appetitive conditioned instrumental behaviours. PMID- 11886453 TI - Task-dependent and cell-type-specific Fos enhancement in rat sensory cortices during audio-visual discrimination. AB - Attention modulates neural activities in sensory cortices. Because cortical neurons are composed of many types of neurons, the activities of these different types of cells can exhibit different modifications depending on whether an animal pays attention to a particular sensory stimulus or not. In the present study, we examined which types of cortical neurons change their activities in rats during one of two types of audio-visual discrimination (AVD) tasks by using Fos immunohistochemistry. In the tasks, both auditory and visual stimuli were simultaneously presented but only one of the two modalities was task-relevant. Once the rats had learned one of the AVD tasks, presenting only relevant sensory stimuli was sufficient for them to perform the task correctly. These results suggest that the rats indeed attended to the relevant stimuli during the performance of the tasks. We found that Fos expression in the primary auditory and visual cortices was enhanced in a task-dependent manner during the performance of the AVD tasks. The enhancement of Fos expression depended on the behavioural significance of the stimulus in the tasks. Moreover, using double immunohistochemistry of Fos and a cell type-specific marker protein (phosphate activated glutaminase, nonphosphorylated neurofilament protein, parvalbumin, calretinin or somatostatin), the task-dependent Fos expression was observed preferentially in excitatory neurons but not in inhibitory interneurons. These results suggest that modulation in cortical excitatory neurons might have critical roles in selecting and processing behaviourally relevant sensory stimuli. PMID- 11886455 TI - Activation of extracellular signal-regulated kinases 1 and 2 by depolarization stimulates tyrosine hydroxylase phosphorylation and dopamine synthesis in rat brain. AB - Production of dopamine is regulated via phosphorylation of tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of catecholamines. Here we have used a preparation of rat striatal slices to examine the involvement of two mitogen-activated protein kinases (MAPKs), extracellular signal-regulated protein kinases 1 and 2 (ERK1/2), in the depolarization-dependent regulation of TH phosphorylation and dopamine synthesis. Depolarization with elevated KCl (45 mm) caused an increase in the phosphorylation state and, thereby, activation of ERK1/2. The same stimulus also increased TH phosphorylation at Ser19, Ser31 and Ser40 (measured using site- and phospho-specific antibodies) and TH activity [measured as 3,4-dihydroxyphenylalanine (DOPA) accumulation]. A MAPK/ERK kinase inhibitor, PD098059, decreased the basal levels of phospho-ERK1/2 and prevented the increase in ERK1/2 phosphorylation induced by depolarization. PD098059 also decreased both basal and depolarization-induced phosphorylation of TH at Ser31 and reduced the increase in Ser40 phosphorylation induced by high potassium, but did not affect Ser19 phosphorylation. PD098059 alone inhibited basal TH activity and decreased the accumulation of DOPA induced by depolarization. These data provide evidence for the involvement of ERK1/2 in the regulation of the state of phosphorylation of TH at Ser31 and Ser40 and a correlation between ERK1/2 dependent phosphorylation of TH and stimulation of dopamine synthesis in the brain. PMID- 11886456 TI - c-Fos expression in dopaminergic and GABAergic neurons of the ventral mesencephalic tegmentum after paradoxical sleep deprivation and recovery. AB - Evidence suggests that dopaminergic neurons of the ventral mesencephalic tegmentum (VMT) could be important for paradoxical sleep (PS). Here, we examined whether dopamine (DA) and adjacent gamma-aminobutyric acid (GABA)-synthesizing neurons are active in association with PS recovery as compared to PS deprivation or control conditions in different groups of rats by using c-Fos expression as a reflection of neural activity, combined with dual immunostaining for tyrosine hydroxylase (TH) or glutamic acid decarboxylase (GAD). Numbers of TH+/c-Fos+ neurons in the substantia nigra (SN) were not significantly different across groups, whereas those in the ventral tegmental area (VTA) were significantly different and greatest in PS recovery. Numbers of GAD+/c-Fos+ neurons in both VTA and SN were greatest in PS recovery. Thus, DA neuronal activity does not appear to be suppressed by local GABAergic neuronal activity during PS but might be altered in pattern by this inhibitory as well as other excitatory, particularly cholinergic, inputs such as to allow DA VTA neurons to become maximally active during PS and thereby contribute to the unique physiological and cognitive aspects of that state. PMID- 11886458 TI - Haemophilia A: effects of inhibitory antibodies on factor VIII functional interactions and approaches to prevent their action. AB - Factor VIII (FVIII) is an essential component of the intrinsic pathway of blood coagulation. Normal functioning of FVIII requires its interactions with other components of the coagulation cascade. In the circulation, it exists as a complex with von Willebrand factor (vWF). Upon activation by thrombin or activated factor X (FXa), activated FVIII (FVIIIa) functions as a cofactor for the serine protease factor IXa. Their complex assembled on the phospholipid surface activates FX to FXa, which consequently participates in formation of thrombin, the key protease of the coagulation cascade. Genetic deficiency in FVIII results in a coagulation disorder haemophilia A, which is treated by infusions of FVIII products. Approximately 25-30% of patients develop antibodies inhibiting FVIII activity (FVIII inhibitors). The major epitopes of inhibitors are located within the A2, C2 and A3 domains of the FVIII molecule. The inhibitory effects of antibodies are manifested at various stages of the FVIII functional pathway, including FVIII binding to vWF, activation of FVIII by thrombin, and FVIIIa incorporation into the Xase complex. We summarize the current knowledge of the FVIII sites involved in interaction with its physiological ligands and different classes of inhibitory antibodies and describe their inhibitory mechanisms. We outline the strategies aimed to overcome the effects of inhibitory antibodies such as development of human/porcine FVIII molecules, resistant to inhibitors. We also discuss approaches to modulate the antibody response, as well as efforts to develop a long-term immunotolerance to FVIII protein. PMID- 11886457 TI - Transient forebrain ischemia induces persistent hyperactivity of large conductance Ca2+-activated potassium channels via oxidation modulation in rat hippocampal CA1 pyramidal neurons. AB - The present study examined temporal changes in activity of large conductance, Ca2+-activated potassium (BKCa) channels in postischemic CA1 pyramidal neurons at 2, 6, 24 and 48 h after reperfusion. These changes in activity and possible cellular mechanisms were examined using the inside--out configuration of patch clamp. The unitary conductance of postischemic BKCa channels increased transiently to 119% of the control at 2 h after reperfusion, and recovered to the control level thereafter. A persistent increase in [Ca2+]i sensitivity of BKCa channels was observed in postischemic CA1 neurons with the maximal sensitivity to [Ca2+]i at 6 h after reperfusion while channel voltage- dependence showed no obvious changes. Kinetic analyses showed that the postischemic enhancement of BKCa channel activity was due to longer open times and shorter closed times as there was no significant changes in opening frequency after ischemia. Glutathione disulphide markedly increased BKCa channel activity in normal CA1 neurons, while reducing glutathione caused a decrease in BKCa channel activity by reducing the sensitivity of this channel to [Ca2+]i in postischemic CA1 neurons. Similar modulatory effects on postischemic BKCa channels were also observed with another redox couple, DTNB and DTT, suggesting an oxidation modulation of BKCa channel function after ischemia. The present results indicate that a persistent enhancement in activity of BKCa channels, probably via oxidation of channels, in postischemic CA1 pyramidal neurons may account for the decrease in neuronal excitability and increase in fAHP after ischemia. The ischemia-induced augmentation in BKCa channel activity may be also associated with the postischemic neuronal injury. PMID- 11886459 TI - The haemophilic pseudotumour. AB - The management of the patient with a haemophilic pseudotumour is complex and carries a high rate of potential complications. There are a number of therapeutic alternatives for this dangerous condition: embolization, radiation, percutaneous management, surgical removal and exeresis, and filling of the dead cavity. It is hoped that with the advent of widespread maintenance therapy, pseudotumours will be less common in the future. It is important that they are diagnosed early, and prevention of muscular haematomas is key to reducing their incidence. Untreated, proximal pseudotumours will ultimately destroy soft tissues, erode bone and may produce neurovascular complications. Surgical excision is the treatment of choice but should only be carried out in major haemophilia centres by a multidisciplinary surgical team. PMID- 11886460 TI - Large FVIII gene deletion confers very high risk of inhibitor development in three related severe haemophiliacs. AB - Haemophilia A displays a broad heterogeneity of genetic defects and of clinical severity. Inhibitor development is the main complication of replacement therapy in severe cases and most patients with inhibitors have gross gene rearrangement or point mutations, which hamper the production of normal circulating factor VIII (FVIII). We have investigated three related severe haemophilia A patients, all of whom have high titre inhibitors. By using long-range polymerase chain reaction (PCR) for FVIII gene inversion, we observed an unusual pattern in these patients. We therefore decided to screen the whole FVIII gene by conformation-sensitive gel electrophoresis. A large FVIII gene deletion spanning exon 2 to exon 25 was identified and we were able to obtain a 18.5 kb PCR product, which is specific for this mutation and useful for determining the carrier state in this family. All three haemophiliacs carrying this very large gene deletion show similar clinical history and very high-titre inhibitors, supporting the observation that inhibitor development seems to be an inherited characteristic. On the basis of our observations we think that this subgroup of patients at very high risk of inhibitor development should be identified by mutation analysis whenever possible, before the beginning of replacement therapy. PMID- 11886461 TI - The effect of resistance training on the frequency of bleeding in haemophilia patients: a pilot study. AB - The benefits and feasibility of progressive resistance training on muscle strength and bleeding profile were studied prospectively in two patients with severe haemophilia. Additionally, retrospective data were collected from three patients who had been training for 11-21 years (one patient for 21 years and two patients for 11 years). Muscle strength increased, especially in muscle groups surrounding the target joints (elbow and knee). Bleeding frequency decreased from 2-3 times per week to 1-2 times per week. Patients who had been training for > 11 years reported bleeding episodes of 2-4 times per month prior to training, but after > 11 years of progressive training a marked decrease in bleeding occurred, as well as a decrease in severity. These data support the importance of resistance training for haemophilia patients, not only for increasing muscle strength, but also for decreasing the frequency and severity of bleeding episodes and the associated pain. A controlled study, with a greater number of patients, is needed to confirm the suggested benefits of resistance training in haemophilia patients. PMID- 11886462 TI - Acquired anti-FVIII inhibitors in children. AB - Acquired inhibitors to FVIII (anti-FVIII) are uncommon in children. An acquired anti-FVIII developed in a previously healthy 4-year-old boy treated with penicillin for streptococcal pharyngitis. Aspirin prophylaxis begun for suspected rheumatic fever led to compartment syndromes of all four extremities, which resolved with high-dose FVIII and surgical decompression. Anti-FVIII in this patient, and the five additional cases identified in a survey of 160 haemophilia treatment centres, occurred at a median age of 8 years, with median initial and peak titres of 4.6 and 6.9 Bethesda Units (BU), respectively. All six presented with bleeding, including haematomas (three intramuscular, one intracranial), and ecchymoses in three. The median baseline FVIII was 0.05 U mL(-1), and the median baseline activated partial thromboplastin time (APTT) was 79.8 s. The inhibitor resolved completely in five patients (83%) within a median 5 months, after treatment with FVIII concentrate, steroids, cytoxan, methotrexate, and no treatment. The inhibitor persisted in the patient with Goodpasture's disease, despite steroids, cytoxan, cyclosporin, and intravenous gamma globulin. Aspirin therapy, in two, worsened ongoing bleeding. The association of penicillin-like drugs in this and three other cases in the literature suggest that to avoid potential catastrophic bleeding, it is prudent to obtain an APTT prior to initiating aspirin for suspected rheumatic fever. In conclusion, acquired anti FVIII inhibitors in children may cause severe bleeding, and remit in the majority after FVIII and/or immunosuppressive therapy. PMID- 11886463 TI - Clinical outcomes and resource utilization associated with haemophilia care in Europe. AB - We conducted a multicentre, cross- sectional study of 1042 haemophilia subjects across Europe to compare various health outcomes associated with on-demand vs. prophylactic factor-substitution therapy. Demographic, medical history, and healthcare resource utilization data were analysed along with the number of bleeding events over the past 6 months. Treatment-cost data were also examined to provide preliminary information for future economic studies. A logistic regression analysis, controlling for other statistically significant covariates, showed that patients treated on demand were 3.4 times more likely to have had a joint bleed over the previous 6 months than those treated with prophylaxis. Multiple regression analyses further confirmed these findings, because on-demand subjects had, on average, 5.15 more joint bleeds over the reporting period than patients treated with prophylaxis. Notably, these findings were even more dramatic for younger haemophilia patients when our study sample was stratified by age. Due to the high cost of factor replacement, healthcare costs were significantly higher for subjects treated prophylactically. While hospital costs for prophylaxis subjects were, on average, lower, statistically significant cost savings for prophylactic subjects were not noted. These results suggest that clinicians and health policy decision-makers should consider the advantages of prophylactic therapy for haemophilia patients in formulating treatment protocols and allocating health resources. PMID- 11886465 TI - Carrier detection and prenatal diagnosis in haemophilia in India: realities and challenges. AB - Organizing services for haemophilia in developing countries with few resources is a formidable task. There is wide variation in haemophilia care and management between developing and developed countries. The management of a genetic disorder such as haemophilia becomes difficult in developing countries where scanty resources are allocated mainly to nutrition and infectious diseases as a first priority. In a country such as India, with one billion people and with a wide diversity in cultural, educational and financial conditions, educating people about such diseases is difficult and will take a long time to have an effect on attitudes. Meanwhile, attempts are being made at several centres in India to develop carrier detection and prenatal diagnosis for this disease. Counselling a carrier for detection of her carrier status remains problematic in India because marriages are still largely arranged by the parents of prospective brides and bridegrooms. Hence, the very idea of communicating the carrier status to the bridegrooms' families may lead to cancellation of the marriages, and concealing the carrier status is associated with guilt both for the bride's parents as well as the bride. Prenatal diagnosis and carrier detection in India therefore must be discussed in this context. PMID- 11886464 TI - Quality-of-life differences between prophylactic and on-demand factor replacement therapy in European haemophilia patients. AB - The European Study on the Clinical Outcomes and Resource Utilization associated with Haemophilia Care was designed to compare various health outcomes associated with on-demand and prophylactic factor substitution methods in European haemophilia patients. While the primary objective of this research is to conduct an economic analysis, an important component of this study is to evaluate quality of-life differences that may exist between patients who utilize these two styles of therapy. Quality-of-life research has emerged as a primary measure of health outcomes because it allows the augmentation of traditional clinical indicators of health with data gathered from the patient's perspective. A total of 1033 haemophilia patients from 16 European haemophilia treatment centres were enrolled in this study. The SF-36, a multidimensional quality-of-life instrument, was administered to all participants. This instrument measures eight health-related quality-of-life dimensions: physical functioning, physical role limitations, bodily pain, general health, vitality, social functioning, emotional role limitations, and mental health. All haemophilia subjects enrolled in the study scored significantly lower than the population normative means in the three physical dimensions and in the general health dimension. HIV-negative haemophiliac subjects differed significantly by factor substitution type in a multivariate analysis examining all eight health dimensions. Univariate analyses testing each dimension separately indicated that patients treated prophylactically reported significantly less bodily pain, better general health, and scored significantly higher in the physical functioning, mental health, and social functioning dimensions. While these results suggest that health-related quality-of-life may be better for haemophilia patients treated prophylactically, future prospective studies that gather periodic quality-of-life data over time should be conducted. PMID- 11886466 TI - Thrombotic stroke associated with the use of porcine factor VIII in a patient with acquired haemophilia. AB - Porcine factor VIII (pFVIII), which is used to control bleeding in patients with congenital or acquired haemophilia who have high-titre neutralizing antibodies to human FVIII, is not known to increase the risk of arterial or venous thrombosis. We have recently encountered a patient with acquired haemophilia who developed a thrombotic left middle cerebral artery distribution stroke while being treated with pFVIII. To our knowledge, this is the first such reported thrombotic event. We speculate that platelet activation induced by pFVIII may have contributed to thrombosis and suggest that pFVIII be used with caution in elderly patients with pre-existing cardiovascular risk factors. PMID- 11886467 TI - Postoperative hepatic laceration in a child with type 3 von Willebrand disease. AB - We report an unusual postoperative complication (hepatic laceration) in a child with type 3 vWD, who has not received any prophylaxis. PMID- 11886468 TI - Danazol therapy in factor X deficiency: more questions than answers. PMID- 11886469 TI - Critical effect of Helicobacter pylori infection on the effectiveness of omeprazole for prevention of gastric or duodenal ulcers among chronic NSAID users. AB - BACKGROUND: The recently reported OMNIUM and ASTRONAUT NSAID ulcer prevention trials using omeprazole to prevent endoscopic ulcer recurrence among chronic NSAID users suggested superiority over misoprostol or ranitidine. AIM: To test the hypothesis the results from the OMNIUM and ASTRONAUT studies would not be generalizible as ulcer healing and ulcer recurrence would differ in relation to Helicobacter pylori status. METHODS: The data regarding H. pylori status were made available by AstraZenca allowing separate analysis of the outcome of those with NSAID ulcers (i.e. without H. pylori infection) and those NSAID use was complicated with the presence of an active H. pylori infection. RESULTS: Reanalysis confirmed that omeprazole was superior to placebo for the prevention of ulcer recurrence in chronic NSAID users. However, overall omeprazole was not significantly better than the subtherapeutic dose (400 microg/day) of misoprostol (14.5% vs. 19.6%, respectively, p =.93); 400 microg of misoprostol was actually superior to omeprazole for the prevention of gastric ulcers among those NSAID ulcers (8.2% vs. 16.6% for misoprostol and omeprazole, respectively; p <.05). Omeprazole was also not statistically different from misoprostol for gastric ulcer prevention in those whose NSAID use was complicated by an active H. pylori infection. Omeprazole was not significantly different from 300 mg of ranitidine for the prevention of NSAID gastric ulcers (14.6% vs. 11.6%, respectively, p =.56). Duodenal ulcers were over represented among H. pylori infected NSAID users and duodenal ulcer prevention was more sensitive to acid suppression than gastric ulcer. CONCLUSION: The OMNIUM and ASTRONAUT trials may have provided an unrealistic sense of security regarding the effectiveness of omeprazole for protection against ulcer recurrence in chronic NSAID users. PMID- 11886470 TI - Effects of selective cyclooxygenase-2 inhibitor and non-selective NSAIDs on Helicobacter pylori-induced gastritis in Mongolian gerbils. AB - BACKGROUND: It is still a point of controversy whether Helicobacter pylori infected patients are more likely to develop mucosal damage while taking NSAIDs. Selective cyclooxygenase (COX-2) inhibitors may be associated with less severe gastric mucosal damage than conventional NSAIDs, but this association is undefined in H. pylori-induced gastritis. The aim of this study was to evaluate the effects of selective COX-2 and nonselective NSAIDs on H. pylori-induced gastritis. METHODS: After intragastric administration of indomethacin, NS-398 or vehicle alone, once daily for 5 days in H. pylori-infected and uninfected Mongolian gerbils, we evaluated gastric mucosal damage, inflammatory cell infiltration and prostaglandin E2 (PGE2) concentration. We investigated whether H. pylori infection induced the COX-2 expression. RESULTS: In H. pylori uninfected groups, the indomethacin-treated group showed the highest mucosal damage score and the lowest PGE2 concentration. There was no difference in mucosal damage scores and PGE2 concentration between NS-398 and vehicle-alone treated group. In H. pylori-infected groups, there was no difference in mucosal damage scores, irrespective of the type of drugs administered. The indomethacin treated group showed the lowest PGE2 concentration, similar to that of the NS-398 and vehicle-alone treated groups, both without H. pylori infection. Gastric neutrophil and monocyte infiltration scores were higher in H. pylori-infected groups than in uninfected groups. However, there was no difference in these scores according to the type of drugs administered, within H. pylori-infected or uninfected groups. COX-2 protein expression was observed in H. pylori-infected Mongolian gerbils but not in uninfected ones. CONCLUSIONS: Our animal study showed that H. pylori infection induced COX-2 expression and increased prostaglandin concentration. Administration of NSAIDs decreased the prostaglandin concentration, but did not increase mucosal damage in H. pylori-induced gastritis. Selective COX-2 inhibitors, instead of conventional NSAIDs, had no beneficial effect on preventing mucosal damage in H. pylori-induced gastritis. PMID- 11886471 TI - Decreased adherence of cagG-deleted Helicobacter pylori to gastric epithelial cells in Japanese clinical isolates. AB - BACKGROUND: The cag pathogenicity island (cag PAI) is a major virulence factor. The ability of Helicobacter pylori to adhere to gastric epithelial cells is an important initial step for virulence. The aim of this study was to evaluate the relationship between genetic variations of cag PAI in Japanese clinical isolates and the ability of H. pylori to adhere to gastric epithelial cells. MATERIALS AND METHODS: The polymerase chain reaction and Southern blot analysis were used to verify the presence or absence of cagA, cagE, cagG, cagI and cagM in the cag PAI in 236 Japanese clinical isolates. The ability of H. pylori to adhere to KATOIII cells was examined by flow cytometry. RESULTS: Seven (3.0%) cag PAI partial deleted strains were found in 236 clinical isolates, and these strains showed three patterns in the deleted region within the cag PAI. All of the cagG-deleted strains showed decreased adherence to KATOIII cells, in comparison with cagG positive strains. These strains had abolished IL-8 induction despite the presence of cagE, which is essential for IL-8 induction. CONCLUSIONS: Our results suggest that cagG or surrounding genes in the cag PAI has a function related to adhesion to epithelial cells. PMID- 11886472 TI - Antibacterial activities of beta-lactamase inhibitors associated with morphological changes of cell wall in Helicobacter pylori. AB - BACKGROUND: Recent study has demonstrated that beta-lactamase inhibitors including clavulanate, sulbactam and tazobactam have an vitro antibacterial effect on Helicobacter pylori. Here we describe the relationship between viability and cell profiles of H. pylori exposed to beta-lactamase inhibitors and some antibiotics in a short-time course. MATERIALS AND METHODS: The antibacterial effects of beta-lactamase inhibitors including clavulanate, sulbactam and tazobactam on the bacterial viability of and morphological changes in H. pylori ATCC43504 were examined. RESULTS: The beta-lactamase inhibitors such as clavulanate and sulbactam alone decreased the viable counts of H. pylori, depending on the antibiotic concentrations. Exposure to these beta-lactamase inhibitors resulted in morphological changes of cell shape, cell-wall disintegration and cell lysis. Among these beta-lactamase inhibitors, clavulanate was the most active, causing a decrease in viable counts and morphological changes such as short filamentous to sphaeroplast formation and lysis. One x minimum inhibitory concentration (MIC) of amoxicillin plus 1 x MIC of clavulanate decreased viable counts effectively compared with 1 x MIC of amoxicillin or 1 x MIC of clavulanate alone, and induced morphological changes of cell shape and cell wall. CONCLUSION: Our results suggest that the beta-lactamase inhibitors alone have concentration-dependent antibacterial activities against H. pylori and affect the morphology of the cell shape and the cell wall in vitro. PMID- 11886474 TI - Increased primary resistance to recommended antibiotics negatively affects Helicobacter pylori eradication. AB - OBJECTIVE: To evaluate the efficacy of two commonly employed treatments for Helicobacter pylori infection and the impact of bacterial resistance to antibiotics on eradication rate. METHODS: Ninety-two consecutive H. pylori positive patients with active peptic ulcer disease were randomly enrolled to receive a 7-day treatment with either lansoprazole 30 mg plus amoxicillin 1 g and clarithromycin 500 mg [all twice a day (b.i.d.), Group A, n = 46]; or bismuth subcitrate 125 mg four times a day (q.i.d.) plus tetracycline 500 mg q.i.d and furazolidone 200 mg b.i.d. (Group B, n = 46) H. pylori status was reassessed 30 days after completion of the therapy and bacterial resistance to the antibiotics was investigated using an in vitro assay. RESULTS: Five patients from each study group were lost to follow up. Both treatments resulted in similar H. pylori eradication rate: 66-60% (per protocol), 59-52% (intention-to-treat) in Groups A and B, respectively (non significant). However, eradication improved to 79% in the absence of H. pylori resistance to clarithromycin or amoxicillin. CONCLUSION: Primary resistance to clarithromycin or amoxicillin may underscore a potentially serious problem for the eradication of H. pylori infection. Testing for bacterial resistance may become necessary to improve therapeutic efficacy. PMID- 11886473 TI - Antral nodularity and positive CagA serology are distinct and relevant markers of severe gastric inflammation in children with Helicobacter pylori infection. AB - BACKGROUND: The aim of this study was to assess whether the endoscopic finding of antral nodularity and serum IgG antibodies to CagA are associated with higher grades of gastric inflammation. MATERIALS AND METHODS: The comprehensive data of two previously published trials were reanalysed. One hundred and fifty-three children (median age 9.5 years) who underwent gastroscopy were included. Biopsy specimens from the antrum and corpus were taken to assess Helicobacter pylori status, gastritis score and lymphoid follicles. During endoscopy, antral nodularity was noted. Serum samples were assayed for IgG antibodies to CagA. RESULTS: The presence of antral nodularity (nod+) and positive CagA serology (CagA+) were each found in 32 of the 77 (41.5%) children who had evidence of H. pylori infection. Cross tabulation showed that 20 children (26%) were nod+/CagA+, 12 (15.5%) nod+/CagA-, 12 (15.5%) nod-/CagA+ and 33 (43%) nod-/CagA-. Gastritis score was significantly lower in nod-/CagA- children than in nod+/CagA- (p =.004), nod-/CagA+ (p =.002) and nod+/CagA+ (p <.001), both in the antrum and corpus. Completely normal gastric histology was only found in the nod-/CagA- subgroup of H. pylori-infected children (eight of 33, 24%). Regression analysis showed that antral nodularity and positive CagA serology were related to severe gastric inflammation independently of each other and age. Separate analysis showed that inflammation (p <.001), activity (p <.001) and H. pylori density (p =.002) scores were significantly lower in nod-/CagA- children compared with nod+/CagA+ children. The number of lymphoid follicles in the gastric mucosa was related to antral nodularity (p =.003) and positive CagA serology (p =.043), independently of each other. CONCLUSIONS: Antral nodularity and positive CagA serology are distinct and relevant markers of severe gastric inflammation in children with H. pylori infection. The lack of both findings in the same child reflects low-grade or no gastritis. PMID- 11886475 TI - What to do about Helicobacter pylori? A decision analysis of its implication on public health. AB - OBJECTIVES: Public health measures to eradicate Helicobacter pylori in the general population may prevent the occurrence of nonulcer dyspepsia (NUD), peptic ulcer (PUD) and gastric cancer, but may at the same time increase the prevalence of gastroesophageal reflux disease (GERD). A decision analysis is carried out to quantify the counteracting influences of H. pylori and resolve the controversy about a public policy to eliminate H. pylori from the general population. METHODS: A compartment model is structured to analyze the jointly beneficial and adverse effects of H. pylori. Gastric acid, H. pylori infection, and other pathophysiological mechanisms influence the occurrence of reflux disease, peptic ulcer and dyspepsia, which all contribute to the occurrence of upper abdominal symptoms. Each influence is modeled as a separate compartment with various connections to other compartments. The simulation is carried out on an electronic spreadsheet. RESULTS: A decision in favor or against eradication of all H. pylori depends primarily on the relative contribution of reflux disease vs. peptic ulcer and dyspepsia to upper abdominal symptoms in the general population. If reflux related symptoms contribute twice more than peptic ulcer plus dyspepsia to the overall occurrence of abdominal symptoms, a strategy to eradicate H. pylori would actually lower rather than raise public health. Below this threshold such strategy may improve general well-being. In the individual patient infected with H. pylori, it remains beneficial to eradicate H. pylori, irrespective of the symptoms' nature. CONCLUSIONS: Although it is advisable to treat H. pylori infection in the individual patient who comes to medical attention, a general policy directed towards complete elimination of H. pylori from the population would not be beneficial. A compartment model provides a simple yet powerful method to assess complex disease behavior. PMID- 11886476 TI - The presence of immunoglobulins in the gastric juice of patients infected with Helicobacter pylori is related to a reduced secretion of acid. AB - BACKGROUND: It has been reported that treatment with proton pump inhibitors (PPI) leads to partial elimination and suppression of Helicobacter pylori. In theory, since acid is known to denature immunoglobulins, this antibacterial activity of PPI may be due to a reduction in the acid output favouring humoral immunity. MATERIALS AND METHODS: We analysed prospectively fasting gastric juice in 54 consecutive patients attending upper endoscopy for pH and levels of IgG, IgA and IgM. In addition, two antral and two corpus biopsies were obtained and histologically examined for the presence of H. pylori. RESULTS: 41/54 patients were infected with H. pylori. Immunoglobulines in the gastric juice of these patients were found in 25/41 (IgG), 27/41 (IgA), and 29/41 (IgM) patients. There was a highly significant difference in the gastric pH when H. pylori infected patients with measurable IgG, IgA, or IgM were compared with those in whom no immunoglobulines were found (median pH: 6 vs. 2 in each group; p <.001) CONCLUSIONS: There is a close correlation between a high gastric pH and the presence of IgG, IgA, and IgM antibodies. Hence, it may be speculated that the efficacy of humoral immunity following H. pylori infection depends on a high pH such as resulting from PPI treatment. PMID- 11886477 TI - Haemochromatosis: understanding the mechanism of disease and implications for diagnosis and patient management following the recent cloning of novel genes involved in iron metabolism. AB - Haemochromatosis, a common recessive genetic disorder in people of Northern European descent, is an iron storage disorder characterized by excessive hepatic iron accumulation resulting from disruption of the regulation of intestinal iron absorption. The identification of novel genes involved in the control of iron absorption from the diet has allowed improved understanding of iron metabolism in health and disease. In particular, the identification of the haemochromatosis gene (HFE) and more recently the transferrin receptor 2 gene (TfR2) together with the specific mutations in these genes which result in hepatic iron overload, has enhanced our understanding of the pathophysiology of haemochromatosis. However, because of the wide variation in phenotypic expression of the disease, there now exists a considerable challenge to diagnosis and patient management. PMID- 11886478 TI - Preventing atherothrombotic events in peripheral arterial disease: the use of antiplatelet therapy. AB - Patients with peripheral arterial disease (PAD) are at increased risk of generalized atherothrombotic events. Epidemiologic data shows a high rate of co prevalence of PAD and atherosclerosis in other vascular beds. Aggressive risk factor modification and antiplatelet therapy has become the cornerstone of treatment to prevent ischaemic events associated with PAD. Recent clinical trials have confirmed the clinical benefit of clopidogrel and ticlopidine in patients with PAD, agents that irreversibly inhibit the binding of adenosine diphosphate to its platelet receptor. In the clopidogrel versus aspirin in patients at risk of ischaemic events (CAPRIE) trial, clopidogrel was associated with an overall risk reduction of 8.7% (compared with aspirin, P=0.043) in myocardial infarction (MI), ischaemic stroke and vascular death. These results demonstrated that long term administration of clopidogrel was effective in preventing ischaemic events in patients with atherosclerotic vascular disease including PAD. Aspirin and/or clopidogrel are the antiplatelet agents of choice for the reduction of atherothrombotic events in patients with PAD. PMID- 11886479 TI - Sleep complaints predict coronary artery disease mortality in males: a 12-year follow-up study of a middle-aged Swedish population. AB - OBJECTIVES: Only a few prospective surveys have been performed to investigate the relationship between sleep complaints and coronary artery disease (CAD) mortality. This study was conducted to determine whether sleep complaints in a middle-aged population predicted total mortality and CAD mortality. DESIGN: A population-based prospective study. Setting. The County of Dalarna, Sweden. SUBJECTS AND METHODS: In 1983, a random sample of 1870 subjects aged 45-65 years responded to a postal questionnaire (response rate 70.2%) including questions about sleep complaints and various diseases. Mortality data for the period 1983 95 were collected, and Cox proportional hazard analyses were used to examine the mortality risks. RESULTS: At 12-year follow-up 165 males (18.2%) and 101 females (10.5%) had died. After adjustment for a wide range of important putative risk factors, difficulties initiating sleep (DIS) were related to CAD death in males [relative risk (RR), 3.1; 95% confidence interval (CI), 1.5-6.3; P < 0.01], but not in females. Short or long sleep duration did not influence risk of CAD mortality or total mortality for either gender. Depression in males increased the risk of death attributed to CAD (RR, 3.0; 95% CI, 1.1-8.4; P < 0.05) and total mortality (RR, 2.2; 95% CI, 1.1-4.5; P < 0.05). CONCLUSION: These results provide evidence that there is an association between difficulties falling asleep and CAD mortality in males. PMID- 11886480 TI - Angiotensin-converting enzyme gene polymorphism in relation to HLA-DR in sarcoidosis. AB - OBJECTIVES: To investigate if an insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme (ACE) gene associates with HLA-DR alleles previously found to be of prognostic interest in Scandinavian sarcoidosis patients. This may contribute to characteristics associated with these HLA-DR alleles, such as a good (DR17) or poor (DR14 or 15) prognosis. DESIGN, SETTINGS AND SUBJECTS: Polymerase chain reaction (PCR) was used for analysing an I/D polymorphism in the gene coding for ACE in 138 subjects; 65 controls and 73 sarcoidosis patients, and for HLA-DR genotyping 67 patients. Serum ACE level (S ACE) was measured in all controls and 72 patients. Sixty-one patients were classified as chronic or nonchronic after 2 years follow-up. All patients were recruited and followed at our outpatient clinic. RESULTS: No significant differences in ACE alleles or genotypes were seen between controls and patients or between patients positive and negative for DR17 or DR14/15. The ACE genotype did not differ between nonchronic and chronic patients. The ACE genotype tended to influence the S-ACE in patients, whilst in controls S-ACE significantly differed between the ACE genotypes. CONCLUSION: This study does not support an association between ACE genotype and sarcoidosis or disease outcome. However, because significantly (P < 0.001) more DR17 positive (17 of 19) than DR14/15 positive (seven of 26) patients were classified as nonchronic, these results instead strengthen the prognostic importance of HLA-DR alleles in Scandinavian sarcoidosis patients. PMID- 11886481 TI - Transforming growth factor (TGF)-beta1 inversely related to vascular cell adhesion molecule-1 in postmenopausal women with coronary artery disease. A possible mechanism for the putative cardioprotective role of TGF-beta1? AB - OBJECTIVE AND DESIGN: Transforming growth factor beta (TGF-beta1) is involved in a variety of physiological processes as well as in many diseases. Both in vitro and in vivo evidence suggest that TGF-beta1 may influence atherogenesis and a dominant protective role of TGF-beta1 on coronary arteries has been proposed. On the other hand, increased levels of soluble adhesion molecules have been found in patients with atherosclerosis, and adhesion of monocytes to the endothelium followed by migration to the intima, has been proved to be an early event in atherosclerosis. The purpose of the present investigation was to look at a possible relationship between circulating active TGF-beta1 and adhesion molecules in postmenopausal women with angiographically verified coronary heart disease (CHD) (n=118). RESULTS: Serum levels of the active form of TGF-beta1 showed a tendency to be lower in patients with increasing number of vessels with more than 50% stenosis (P=0.058), and there was higher TGF-beta1 in the group with one vessel disease compared with those with two or more vessels affected (P=0.041). Additionally, negative association between TGF-beta1 and VCAM-1 was found (r= 0.26, P=0.023). However, no associations were observed between TGF-beta1 and intercellular adhesion molecule-1 (ICAM-1) or E-selectin in the present study. CONCLUSION: We observed an inverse correlation between the active form of TGF beta1 and VCAM-1 in postmenopausal women with verified CHD. These results may suggest a role of TGF-beta1 in CHD. PMID- 11886482 TI - Natriuretic peptides during the development of doxorubicin-induced left ventricular diastolic dysfunction. AB - OBJECTIVES: To investigate changes in plasma atrial natriuretic peptide (ANP), N terminal pro-atrial natriuretic peptide (NT-pro-ANP) and brain natriuretic peptide (BNP) during the development of doxorubicin-induced left ventricular systolic and diastolic dysfunction as measured by echocardiography (ECHO). DESIGN: Prospective study. SETTING: University hospital. SUBJECTS: Twenty-eight adult patients with non-Hodgkin's lymphoma, who received doxorubicin to the cumulative dose of 400-500 mg m(-2). MAIN OUTCOME MEASURES: The relationship between plasma natriuretic peptides and systolic and diastolic ECHO indices after the cumulative doxorubicin doses of 200, 400 and 500 mg m(-2). RESULTS: Left ventricular ejection fraction (LVEF, by 2D ECHO) decreased from 58 +/- 1.7 to 52.5 +/- 1.3% (P=0.036) and fractional shortening (FS) from 34.6 +/- 1.4 to 27.8 +/- 0.9% (P=0.002). Peak E wave velocity decreased from 63.3 +/- 3.2 to 51.3 +/- 2.6 cm s(-1) (P=0.008) resulting in a statistically nonsignificant decrease in E/A ratio from 1.08 +/- 0.01 to 0.85 +/- 0.07. A significant decrease was observed in the percentage of left ventricular filling during the 1/3 of diastole (1/3FF) from 42.2 +/- 1.7 to 36.5 +/- 2.0% (P < 0.001). LV end systolic diameter increased from 32 +/- 1 to 38 +/- 1 mm (P=0.011), whereas left atrial (LA) diameter remained unchanged. Peak filling rate decreased from 4.4 +/- 0.2 to 4.0 +/- 0.2 stroke volume s(-1) (SV s(-1)) (ns). Plasma levels of ANP increased from 16.4 +/- 1.3 to 22.7 +/- 2.4 pmol L(-1) (P=0.002), NT-pro-ANP from 288 +/- 22 to 380 +/- 42 pmol L(-1) (P=0.019) and BNP from 3.3 +/- 0.4 to 8.5 +/- 2.0 pmol L( 1) (P=0.020). There was a significant inverse correlation between the decrease in FS and the increases in plasma NT-pro-ANP (r= -0.524, P=0.018) and plasma BNP (r=0.462, P=0.04) and between the decrease in PFR and the increases in plasma ANP (r= -0.457, P=0.043) and plasma NT-pro-ANP (r= -0.478, P=0.033). Furthermore, after doxorubicin therapy, significant inverse correlations were observed between E/A ratio and plasma ANP (r= -0.535, P=0.008), between E/A ratio and plasma NT pro-ANP (r= -0.432, P=0.04) and between E/A ratio and plasma BNP (r= -0.557, P=0.006) as well as between 1/3FF and plasma BNP (r= -0.493, P=0.017). There was also a trend for correlation between LA diameter and plasma BNP (r=0.395, P=0.062) and peak E wave velocity and plasma BNP (r= -0.414, P=0.05), respectively. However, no significant correlations were observed between any of the systolic parameters and natriuretic peptide levels. CONCLUSIONS: The results of this prospective study show that during the evolution of doxorubicin-induced LV dysfunction the secretion of natriuretic peptides is more closely associated with the impairment of left ventricular diastolic filling than with the deterioration of LV systolic function. PMID- 11886483 TI - Exploring sex differences in case fatality in acute myocardial infarction or coronary death events in the northern Sweden MONICA Project. AB - OBJECTIVES: To investigate sex differences in reaching diagnosis, medical management and case fatality (CF) in acute myocardial infarction (AMI) in the population aged 35-64 years in northern Sweden. METHODS: Within the framework of the World Health Organization Multinational Monitoring of Trends and Determinants in Cardiovascular Diseases (MONICA) Project, definite AMI was monitored in people aged 35-64 years from 1989 through 1995 (target population 510 000 in 1991). SETTING: In a population based coronary register, all coronary events were recorded in nine hospitals in 1989-95. RESULTS: The number of events included in the definite coronary myocardial infarction register was 2483 men and 669 women. On admission, a higher proportion of men with definite AMI had chest pain or ECG changes typical for AMI (P < 0.0001). Disagreement between clinical diagnosis and classification by MONICA criteria occurred more often in women (P=0.008). A significantly higher proportion of men was admitted in the coronary care unit and they were significantly more often treated with thrombolytics, nitroglycerine, beta-blockers, or antiplatelet agents. Women received significantly more diuretics, inotropics or calcium antagonists. Diabetes, conferring a worse prognosis, was more common in women (20 vs. 15%; P=0.003). Prehospital CF was significantly higher in men (24.1 vs. 18.3%; P=0.005), but in patients treated in hospital, the CF was significantly lower in men (12.7 vs. 21.2%; P < 0.001). Total CF was equal in men and women. CONCLUSIONS: Several factors contributing to the excess in-hospital CF in women were identified, including greater problems in diagnosis of AMI in women which may be one of the reasons for less intensive treatment in women. Differences in co-morbidity, most notably diabetes and medical treatment between men and women with acute AMI may also have played a part. PMID- 11886484 TI - Oral oestradiol/trimegestone replacement reduces procarboxypeptidase U (TAFI): a randomized, placebo- controlled, 12-week study in early postmenopausal women. AB - OBJECTIVE: To investigate the effects of short-term postmenopausal oral hormone administration on plasma levels of procarboxypeptidase U (proCPU, thrombin activatable fibrinolysis inhibitor, EC 3.4.17.20), an inhibitor of fibrinolysis, in healthy early postmenopausal women. DESIGN: A prospective, randomized, placebo controlled study. SETTING: Outpatient clinic of the Department of Obstetrics and Gynaecology. SUBJECTS: Seventy-seven healthy early postmenopausal women were screened of whom 65 were randomized. Analyses were based on 60 participants. INTERVENTIONS: The women received oral micronized oestradiol 2 mg either alone (E2 group, n=16), or sequentially combined with dydrogesterone 10 mg (E2 + D group, n=14) or trimegestone 0.5 mg (E2 + T, n=14), or placebo (n=16) for 12 weeks. MAIN OUTCOME MEASURE: ProCPU concentrations at baseline, and at 4 and 12 weeks of treatment. RESULTS: Four weeks of E2 + T was associated with a significant decrease in the fasting proCPU concentration, which was sustained after 12 weeks [t=0: 636 +/- 57 U L(-1) (mean +/- SD); t=4: 583 +/- 63UL-1; t=12: 589 +/- 48 U L(-1); ANCOVA versus placebo: P=0.011]. The percentage change from baseline versus placebo in this group was -8.4% [95% confidence interval (CI) 15.7 to -1.1] after 4 weeks and -5.9% (95% CI -11.7 to -0.1) after 12 weeks. There were no significant changes versus placebo in the E2 group nor in the E2 + D group. CONCLUSION: Short-term treatment with E2 + T, but not E2 alone or E2 + D, lowers proCPU concentration. These findings add to accumulating evidence suggesting that different progestagens added to oestrogen replacement may differentially affect the risk of arterial and venous disease. PMID- 11886485 TI - A C-1291G polymorphism in the alpha2A-adrenergic receptor gene (ADRA2A) promoter is associated with cortisol escape from dexamethasone and elevated glucose levels. AB - OBJECTIVES: The objective of the current study was to examine the potential impact of a C right arrow G substitution at position -1291 of the alpha2A adrenergic receptor gene (ADRA2A) promoter on obesity and estimates of insulin, glucose, and lipid metabolism as well as circulating hormones, including salivary cortisol in 284 unrelated Swedish men born in 1944. MAIN OUTCOME MEASURES: The subjects were genotyped by using PCR amplification of the promoter region of the ADRA2A gene followed by digestion with the restriction enzyme MspI. RESULTS: The frequencies were 0.23 for allele C and 0.77 for allele G. The observed genotype frequencies were 45.8 and 54.2% for C/G and G/G, respectively. Heterozygotes (n=121) had significantly (P=0.009) higher salivary cortisol levels after 0.5 mg dexamethasone compared with G/G homozygotes (n=143). Fasting glucose was found to be significantly (P=0.017) higher in heterozygotes than in G/G homozygotes. The latter group had also a borderline significantly (P=0.080) higher mean diastolic blood pressure. These results were all adjusted for the potential confounding effect of body mass index (BMI) and waist-to-hip ratio (WHR). Other measurements such as BMI, WHR, abdominal sagittal diameter, total testosterone, insulin-like growth factor I, serum leptin, fasting insulin and serum lipids were not different across the ADRA2A genotype groups. CONCLUSIONS: In conclusion, we have shown that an C --> G polymorphism at position -1291 of the ADRA2A gene is associated with a subnormal cortisol response to dexamethasone, elevated glucose levels and perhaps increased diastolic blood pressure. The pathophysiology could involve an altered density of the alpha2A-AR that destabilizes the sympathetic hypothalamic-pituitary-adrenal systems in those with genetic vulnerability in the alpha2A-adrenergic receptor gene promoter. PMID- 11886486 TI - Parental age and coronary disease in the general male population. AB - OBJECTIVE: To investigate the role of achieved age in father or mother for long term prognosis in men with respect to fatal and nonfatal coronary events, and mortality from cardiovascular disease, cancer, and all causes. DESIGN: Prospective follow-up study. SETTING: City of Goteborg, Sweden. SUBJECTS: A total of 6242 men aged 51-59 out of 7100 men who took part in the second screening of the Primary Prevention Study in 1974-77 and who had stated the age of both mother and father at death, or their current age, if they were still alive. MAIN OUTCOME MEASURES: Fatal and nonfatal coronary events, and mortality from cardiovascular disease, cancer, and all causes during follow-up until 1996. RESULTS: Of the men (n=2135) whose father had died before the age of 70, 8.4 per 1000 observation years died from coronary disease during follow-up. Coronary mortality decreased continuously with increasing age in the father and in those whose father had achieved an age of 90 or more 3.2 per 1000 years died [hazard ratio (HR) after adjustment for coronary risk factors, socio-economic status, and history of myocardial infarction in either parent, 0.41 (95% CI 0.23-0.73; P for trend <0.0001)]. This was reflected in reduced risk of mortality from any cause (P for trend after adjustment 0.003). No association with cancer death was found. With respect to hospitalization for myocardial infarction men whose father had survived to at least 90 had an adjusted HR of 0.60 (0.40-0.89) compared with men whose father died before 70 (P for trend 0.0006). The effect of achieved age in the mother was weaker and after adjustment trends were weakly significant for death from cardiovascular and all causes (P=0.01 and 0.03, respectively), but not for any other end-point. CONCLUSION: Paternal, but not maternal, longevity appears to protect against coronary disease, by mechanisms that are largely unknown. PMID- 11886487 TI - ANCA-positive periaortic vasculitis: does it fall within the spectrum of vasculitis? AB - Retroperitoneal fibrosis (RPF) is a disease of unknown aetiology that has sometimes been reported in association with connective tissue disorders and systemic vasculitis. We report here two cases of antineutrophil cytoplasmic antibody (ANCA)-positive RPF showing clinical evidence of rapidly progressive glomerulonephritis. Although treatment with prednisone and cyclophosphamide led to a remission of RPF in both cases, renal function was restored in only one patient and the other progressed to chronic renal failure. The paper reviews the literature concerning ANCA-positive RPF and discusses the relationship between ANCA-positive vasculitis and RPF. PMID- 11886488 TI - Systemic granulomatosis and hypercalcaemia following intravesical bacillus Calmette-Guerin immunotherapy. PMID- 11886489 TI - Recurrent pneumonia with unconsciousness. PMID- 11886490 TI - Simple plasma exchange reduced autoantibody to von Willebrand factor-cleaving protease in a Japanese man with ticlopidine-associated thrombotic thrombocytopenic purpura. PMID- 11886493 TI - Steroid sulfatase in the human hair follicle concentrates in the dermal papilla. AB - 5 alpha-dihydrotestosterone is known to play a crucial part in the regulation of hair growth and in the development of androgenetic alopecia. 5 alpha dihydrotestosterone is formed locally within the hair follicle from the systemic precursor testosterone by cutaneous steroid 5 alpha-reductase. Moreover, adrenal steroids such as dehydroepiandrosterone are converted to 5 alpha dihydrotestosterone by isolated hair follicles, which may provide an additional source of intrafollicular 5 alpha-dihydrotestosterone levels. Elevated urinary dehydroepiandrosterone and serum dehydroepiandrosterone sulfate have been reported to be present in balding young men. These reports suggest that dehydroepiandrosterone sulfate may act as an important endocrine factor in the development of androgenetic alopecia. Hence the question arises whether the dehydroepiandrosterone sulfate can be metabolized within the hair follicles to yield dehydroepiandrosterone by the microsomal enzyme steroid sulfatase, and where steroid sulfatase might be localized. We therefore performed immunostaining for steroid sulfatase on human scalp biopsies as well as analysis of steroid sulfatase enzyme activity in defined compartments of human beard and occipital hair follicles ex vivo. Using both methods steroid sulfatase was primarily detected in the dermal papilla. Steroid sulfatase activity was inhibited by estrone-3-O-sulfamate, a specific inhibitor of steroid sulfatase, in a concentration-dependent way. Furthermore, we show that dermal papillae are able to utilize dehydroepiandrosterone sulfate to produce 5 alpha-dihydrotestosterone, which lends further support to the hypothesis that dehydroepiandrosterone sulfate contributes to androgenetic alopecia and that steroid sulfatase inhibitors could be novel drugs to treat androgen-dependent disorders of the hair follicle such as androgenetic alopecia or hirsutism. PMID- 11886492 TI - Heparin-binding epidermal-growth-factor-like growth factor activation of keratinocyte ErbB receptors Mediates epidermal hyperplasia, a prominent side effect of retinoid therapy. AB - Sun-protected human skin was maintained in organ culture and treated with all trans retinoic acid in the presence or absence of reversible or irreversible pharmacologic antagonists of c-erbB receptor tyrosine kinase activity. In the absence of these inhibitors, all-trans retinoic acid induced epidermal hyperplasia comparable to that induced in intact skin by all-trans retinol or all trans retinoic acid itself. There was a strong correlation between inhibition of epidermal hyperplasia in organ culture and inhibition of epidermal-growth-factor dependent keratinocyte growth in monolayer culture. In additional studies it was shown that all-trans retinoic acid could overcome the known inhibitory effects of calcium on expression of HB-EGF-like growth factor mRNA in organ-cultured skin. Further, it was shown that an antibody to HB-EGF-like growth factor inhibited retinoid-stimulated epidermal hyperplasia in organ culture and reduced proliferation in cultured keratinocytes. In contrast, the c-erbB receptor tyrosine kinase antagonists and the neutralizing HB-EGF-like growth factor antibody were ineffective in inhibiting all-trans-retinoic-acid-dependent survival and proliferation of human dermal fibroblasts. Taken together, these data indicate (i) that retinoid-induced epidermal hyperplasia in human skin proceeds through c-erbB, and (ii) that HB-EGF-like growth factor is one of the c erbB ligands mediating this effect. PMID- 11886494 TI - Human follicular papilla cells carry out nonadipose tissue production of leptin. AB - Leptin, a satiety-regulating cytokine, is predominantly expressed by adipocytes, although recently the nonadipose tissue production of leptin has been reported. To investigate the possibility of leptin production by human scalp hair follicles, we examined leptin production and its mRNA expression by cultured human follicular papilla cells. We isolated 12 human follicular papilla cell lines from different individuals. They were identified by their morphology, their high alpha-smooth-muscle actin expression, their inability to differentiate into adipocytes, and by the lack of mRNA for adipose-specific fatty acid binding protein. All the human follicular papilla cell lines, but not neonatal human dermal fibroblasts, produced significant amounts of leptin demonstrable by enzyme linked immunosorbent assay. We demonstrated leptin mRNA expression by human follicular papilla cell lines, but not by neonatal human dermal fibroblasts, by reverse transcription polymerase chain reaction. By immunohistochemistry and in situ hybridization, we detected both leptin protein and mRNA at the lower portion of the hair follicle, i.e., hair matrix, inner root sheath of the hair bulb, and human follicular papilla cells. In contrast, the leptin receptor with intracytoplasmic signal sequence was detected in the follicular papilla cells immunohistochemically, and the long isoform of the leptin receptor mRNA was demonstrated in the human follicular papilla cell lines by reverse transcription polymerase chain reaction. Finally, by using these human follicular papilla cell lines, we showed that cytokines such as interleukin-1 beta, tumor necrosis factor alpha, interferon-gamma, and interleukin-4, and growth factors such as epidermal growth factor, basic fibroblast growth factor, and transforming growth factor beta1, but not vascular endothelial growth factor, hepatocyte growth factor, keratinocyte growth factor, and insulin-like growth factor 1, significantly downregulated the production of leptin. These data demonstrated that human follicular papilla cells produce leptin and express the functional leptin receptor in vivo and in vitro, suggesting its autocrine function. Moreover, the regulation pattern of its production by various factors suggests a pivotal role of leptin in hair biology. PMID- 11886495 TI - Melanocyte-associated T cell epitopes can function as autoantigens for transfer of alopecia areata to human scalp explants on Prkdc(scid) mice. AB - Alopecia areata is a tissue restricted autoimmune condition affecting the hair follicle, resulting in hair loss. The goal of this study was to test the hypothesis that the autoantigen of alopecia areata is melanocyte associated. Potential autoantigens were tested in the human scalp explant/Prkd(scid) CB-17 mouse transfer system. Scalp T cells from lesional (bald) alopecia areata scalp were cultured with antigen-presenting cells, and antigen, along with interleukin 2. The T cells were then injected into autologous lesional scalp grafts on SCID mice, and hair regrowth was measured. Hair follicle homogenate was used as an autoantigen control. T cells cultured with melanoma homogenate induced statistically significant reduction in hair growth (p <0.01 by ANOVA). HLA-A2 restricted melanocyte peptide epitopes were then tested with lesional scalp T cells from HLA-A2-positive alopecia areata patients. Melanocyte-peptide-activated T cells significantly reduced the number of hairs regrowing in two experiments with six patients (p <0.001 by ANOVA). Injected scalp grafts showed histologic and immunochemical changes of alopecia areata. The most consistent peptide autoantigens were the Gp100-derived G9-209 and G9-280 peptides, as well as MART-1 (27-35). Melanocyte peptide epitopes can function as autoantigens for alopecia areata. Multiple peptides were recognized, suggesting epitope spreading. PMID- 11886496 TI - Dimethylfumarate inhibits tumor-necrosis-factor-induced CD62E expression in an NF kappa B-dependent manner. AB - Fumaric acid esters are thought to improve psoriasis by altering leukocyte, keratinocyte, and/or endothelial functions. To determine specificity, kinetics, and molecular mechanisms of different fumaric acid esters in their ability to inhibit endothelial cell activation, we analyzed CD62E and CD54 expression in endothelial cells in vivo and in vitro. In lesional skin of psoriatic patients, oral fumaric acid ester treatment resulted in a marked reduction of CD62E but not CD54 expression on dermal microvessels. Using human umbilical vein endothelial cells, dimethylfumarate almost completely inhibited tumor-necrosis-factor-induced CD62E, but not CD54 expression at concentrations < or = 70 microM, mimicking the situation in vivo. A 60 min dimethylfumarate preincubation was sufficient to block tumor-necrosis-factor-induced CD62E expression for up to 24 h. In contrast, equimolar concentrations of methylhydrogenfumarate, the hydrolysis product of dimethylfumarate, did not suppress tumor-necrosis-factor-induced CD62E expression. Likewise, all fumaric acid esters other than dimethylfumarate were ineffective. Using CD62E, NF-kappa B, or AP-1-responsive promoter constructs, dimethylfumarate inhibited tumor-necrosis-factor-induced activation of the CD62E and the NF-kappa B but not the AP-1 promoter construct. In summary, at a dose range < or = 70 microM, dimethylfumarate appeared to be a specific inhibitor of CD62E expression in an NF-kappa B-dependent manner. PMID- 11886497 TI - Fibrinogen and fibrin are anti-adhesive for keratinocytes: a mechanism for fibrin eschar slough during wound repair. AB - During cutaneous wound repair the epidermis avoids the fibrin-rich clot; rather it migrates down the collagen-rich dermal wound margin and over fibronectin-rich granulation tissue. The mechanism(s) underlying keratinocyte movement in this precise pathway has not been previously addressed. Here we demonstrate that cultured human keratinocytes do not express functional fibrinogen/fibrin receptors, specifically alpha v beta 3. Biologic modifiers known to induce integrin expression or activation did not induce adhesion to fibrin, fibrinogen, or its fragments. Epidermal explant outgrowth and single epidermal cell migration failed to occur on either fibrin or fibrinogen. Surprisingly, fibrin and fibrinogen mixed at physiologic molar ratios with fibronectin abrogated keratinocyte attachment to fibronectin. Keratinocytes transduced with the beta 3 integrin subunit cDNA, expressed alpha v beta 3 on their surface and attached to and spread on fibrinogen and fibrin. beta-gal cDNA-transduced keratinocytes did not demonstrate this activity. Furthermore, beta 3 cDNA-transduced keratinocyte adhesion to fibrin was inhibited by LM609 monoclonal antibody to alpha v beta 3 in a concentration-dependent fashion. From these data, we conclude that normal human keratinocytes cannot interact with fibrinogen and its derivatives due to the lack of alpha v beta 3. Thus, fibrinogen and fibrin are authentic anti adhesive for keratinocytes. This may be a fundamental reason why the migrating epidermis dissects the fibrin eschar from wounds. PMID- 11886498 TI - Keratinocyte-derived granulocyte-macrophage colony stimulating factor accelerates wound healing: Stimulation of keratinocyte proliferation, granulation tissue formation, and vascularization. AB - Chronic, nonhealing wounds represent a major clinical challenge to practically all disciplines in modern medicine including dermatology, oncology, surgery, and hematology. In skin wounds, granulocyte-macrophage colony stimulating factor (GM CSF) is secreted by keratinocytes shortly after injury and mediates epidermal cell proliferation in an autocrine manner. Many other cells involved in wound healing including macrophages, lymphocytes, fibroblasts, endothelial cells, and dendritic cells synthesize GM-CSF and/or are targets of this cytokine. Therefore, GM-CSF is a pleiotropic cytokine evoking complex processes during wound repair. Despite this complexity and the scarcity of mechanistic understanding GM-CSF has been employed in trials of clinical treatment of skin wounds with some success. In this study, we evaluated a transgenic mouse model in order to analyze the effects of an excess of keratinocyte-derived GM-CSF on excisional wound healing in the skin. Transgenic mice constitutively overexpressing GM-CSF in the basal layer of the epidermis displayed accelerated reepithelialization of full thickness skin wounds. In the early stages of wound repair, transgenic mice exhibited significantly higher numbers of proliferating keratinocytes at the wound edges and increased formation of granulation tissue with enhanced neovascularization. As a potential mechanism of these beneficial changes, we identified the differential temporal regulation of cytokines such as transforming growth factor-beta, a known angiogenetic factor, interferon-gamma, a proinflammatory cytokine, and interleukin 6, an essential factor for reepithelialization, in transgenic mice versus controls. We propose that the beneficial effects observed in GM-CSF transgenics are due not only to direct GM CSF action but in addition to indirect processes via the induction of secondary cytokines. PMID- 11886499 TI - Novel and recurrent mutations in the genes encoding keratins K6a, K16 and K17 in 13 cases of pachyonychia congenita. AB - Thirteen patients with pachyonychia congenita types 1 and 2 were studied, two of which had a family history of pachyonychia and 11 of which were sporadic cases. Heterozygous mis-sense or small in-frame insertion/deletion mutations were detected in the genes encoding keratins K6a, K16, and K17 in all cases. Three novel mutations, F174V, E472K, and L469R were found in the K6a gene. Two novel mutations, M121T and L128Q were detected in K16. Similarly, three novel mutations, L95P, S97del, and L99P were found in K17. In addition, we identified recurrent mutations N171del (three instances) and F174S in K6a and R94H in K17. Analysis of both phenotype and genotype data led to the following conclusions: (i) K6a or K16 mutations produce the pachyonychia congenita type 1 phenotype, whereas K17 (or K6b) mutations cause pachyonychia congenita type 2; (ii) the presence of pilosebaceous cysts following puberty is the best indicator of pachyonychia congenita type 2; (iii) prepubescent patients are more difficult to classify due to the lack of cysts; and (iv) natal teeth are indicative of pachyonychia congenita type 2, although their absence does not preclude the pachyonychia congenita type 2 phenotype. This study establishes useful diagnostic criteria for pachyonychia congenita types 1 and 2, which will help limit unnecessary DNA analysis in the diagnosis and management of this genetically heterogeneous group of genodermatoses. PMID- 11886500 TI - Transduction of the E6 and E7 genes of epidermodysplasia-verruciformis-associated human papillomaviruses alters human keratinocyte growth and differentiation in organotypic cultures. AB - Epidermodysplasia-verruciformis-associated human papilloma virus DNA has been detected in skin cancers, in premalignant and benign skin lesions, and in plucked hairs from immunocompetent and immunosuppressed patients. The role of epidermodysplasia-verruciformis-associated human papilloma virus in the pathogenesis of nonmelanoma skin cancer is still enigmatic. In organotypic cultures we investigated the effects of retroviral transduction of the E6 and E7 genes of epidermodysplasia-verruciformis-associated human papilloma virus types 5, 12, 15, 17, 20, and 38 on the growth and differentiation of human keratinocytes. Differentiation was disturbed to different degrees as revealed by histology and by the expression patterns of differentiation markers keratin 10 and small proline rich protein 2. Conversely, proliferating cell nuclear antigen was induced in some of the suprabasal, differentiated cells to varying extent. No unscheduled DNA synthesis was detected in these cells, however, as probed by 5' bromo-2'-deoxyuridine incorporation. Most intriguingly, when the E6 and E7 genes of epidermodysplasia-verruciformis-associated human papilloma virus types 15 and 17 were transduced, a broadening layer of basal cells and an accelerated differentiation were observed. In addition, "papilla-like structures" comprising basal-like keratinocytes arose from the basal layer into the differentiated layers. These cells did not express the differentiation markers keratin 10 and small proline rich protein 2, but did actively replicate DNA. These observations warrant further research by using this system to elucidate the replication strategy of epidermodysplasia-verruciformis-associated human papilloma virus types in keratinocytes and to shed light on the role of these human papilloma virus types in the pathogenesis of skin cancer. PMID- 11886501 TI - Two different mutations in the cytoplasmic domain of the integrin beta 4 subunit in nonlethal forms of epidermolysis bullosa prevent interaction of beta 4 with plectin. AB - The integrin alpha 6 beta 4 plays a crucial role in the assembly and maintenance of hemidesmosomes. Previous work has shown that the recruitment of plectin into hemidesmosomes is dependent on beta 4 and involves a region of the beta 4 cytoplasmic domain, which contains the first two fibronectin (FNIII) repeats and a short region of the connecting segment. Two missense mutations (R1225H and R1281W) in beta 4 that are responsible for nonlethal forms of epidermolysis bullosa are located in the second FNIII repeat. One of them is confined to a loop region that connects two beta strands (EC') whereas the other is located at the N terminal end of the second FNIII repeat. We here report that these mutations render beta 4 unable to interact with plectin and prevent the localization of plectin in hemidesmosomes. Substitution of a lysine residue (K1279W) that forms part of the same loop as R1281 had no effect on the ability of beta 4 to recruit plectin. Furthermore, we show that an extended loop structure in beta 4, composed of the amino acids DDN (1262--1264), which resembles the RGD integrin-binding loop in fibronectin, is not involved in the binding to plectin. These results further demonstrate that binding of beta 4 to plectin is essential for the proper formation and function of hemidesmosomes and that loss of the interaction between beta 4 and plectin is associated with a mild form of epidermolysis bullosa. PMID- 11886502 TI - Protease-activated receptor 2, a receptor involved in melanosome transfer, is upregulated in human skin by ultraviolet irradiation. AB - Previous studies have shown that the protease-activated receptor 2 is involved in skin pigmentation through increased phagocytosis of melanosomes by keratinocytes. Ultraviolet irradiation is a potent stimulus for melanosome transfer. We show that protease-activated receptor 2 expression in human skin is upregulated by ultraviolet irradiation. Subjects with skin type I, II, or III were exposed to two or three minimal erythema doses of irradiation from a solar simulator. Biopsies were taken from nonexposed and irradiated skin 24 and 96 h after irradiation and protease-activated receptor 2 expression was detected using immunohistochemical staining. In nonirradiated skin, protease-activated receptor 2 expression was confined to keratinocytes in the lower one-third of the epidermis. After ultraviolet irradiation protease-activated receptor 2 expression was observed in keratinocytes in the upper two-thirds of the epidermis or the entire epidermis at both time points studied. Subjects with skin type I showed delayed upregulation of protease-activated receptor 2 expression, however, compared with subjects with skin types II and III. Irradiated cultured human keratinocytes showed upregulation in protease-activated receptor 2 expression as determined by immunofluorescence microscopy and Western blotting. Cell culture supernatants from irradiated keratinocytes also exhibited a dose-dependent increase in protease-activated receptor-2 cleavage activity. These results suggest an important role for protease-activated receptor-2 in pigmentation in vivo. Differences in protease-activated receptor 2 regulation in type I skin compared with skin types II and III suggest a potential mechanism for differences in tanning in subjects with different skin types. PMID- 11886503 TI - Regulation of keratin expression by ultraviolet radiation: differential and specific effects of ultraviolet B and ultraviolet a exposure. AB - Skin, the most superficial tissue of our body, is the first target of environmental stimuli, among which is solar ultraviolet radiation. Very little is known about the regulation of keratin gene expression by ultraviolet radiation, however, although (i) it is well established that ultraviolet exposure is involved in skin cancers and photoaging and (ii) keratins represent the major epidermal proteins. The aim of this study was to analyze the regulation of human keratin gene expression under ultraviolet B (290-320 nm) or ultraviolet A (320 400 nm) irradiation using a panel of constructs comprising different human keratin promoters cloned upstream of a chloramphenicol acetyl transferase reporter gene and transfected into normal epidermal keratinocytes. By this approach, we demonstrated that ultraviolet B upregulated the transcription of keratin 19 gene and to a lesser extent the keratin 6, keratin 5, and keratin 14 genes. The DNA sequence responsible for keratin 19 induction was localized between -130 and +1. In contrast to ultraviolet B, ultraviolet A irradiation induced only an increase in keratin 17, showing a differential gene regulation between these two ultraviolet ranges. The induction of keratin 19 was confirmed by studying the endogenous protein in keratinocytes in classical cultures as well as in skin reconstructed in vitro and normal human skin. These data show for the first time that keratin gene expression is regulated by ultraviolet radiation at the transcriptional level with a specificity regarding the ultraviolet domain of solar light. PMID- 11886504 TI - Activators of peroxisome proliferator-activated receptors protect human skin from ultraviolet-B-light-induced inflammation. AB - Peroxisome proliferator-activated receptors (PPAR) are members of a nuclear receptor superfamily, which were initially described in the context of fatty acid degradation and adipocyte differentiation. In this study we tested the hypothesis that peroxisome proliferator-activated receptor activation also controls inflammation. In an in vitro model with human keratinocytes inflammation was mimicked by irradiation with ultraviolet B light (150 mJ per cm(2)). Activators for PPAR-alpha (WY-14,643, clofibrate) were shown to reverse ultraviolet-B-light mediated expression of inflammatory cytokines (interleukin-6, interleukin-8). An activator preferentially for PPAR-beta (bezafibrate) did not show prominent effects on interleukin-6 and interleukin-8 expression. The anti-inflammatory action of WY-14,643 on skin cells was further demonstrated by in vivo testings in which topically applied WY-14,643 markedly increased the minimal erythema dose in ultraviolet-B-irradiated skin. Additionally, it was shown that ultraviolet B irradiation led to a decrease of all three peroxisome proliferator-activated receptor subsets at the mRNA level. Also transactivation of peroxisome proliferator response element was attenuated by ultraviolet B irradiation. The downregulation of peroxisome proliferator-activated receptors by ultraviolet B irradiation provides a possible mechanism that leads to exaggerated and prolonged inflammation. This work suggests the possibility of PPAR-alpha activators as novel nonsteroidal anti-inflammatory drugs in the topical treatment of common inflammatory skin diseases such as atopic dermatitis, psoriasis, and photodermatitis. PMID- 11886505 TI - Protection against pyrimidine dimers, p53, and 8-hydroxy-2'-deoxyguanosine expression in ultraviolet-irradiated human skin by sunscreens: difference between UVB + UVA and UVB alone sunscreens. AB - As DNA damage induced by ultraviolet radiation plays an essential role in skin cancer induction, we pursued the measure of several DNA lesions induced by ultraviolet radiation in human skin for determining the efficacy of different topical photoprotectors. Non-exposed skin (buttocks from 20 individuals) was exposed to 10 doses of ultraviolet, which corresponded to three to four minimal erythema doses of solar-simulating radiation, and biopsies were taken at 24 h within the half and one minimal erythema dose sites and a nonirradiated, adjacent control area. We report that even suberythemal doses of ultraviolet radiation are capable of inducing substantial DNA damage, namely pyrimidine dimers, p53 induction, and the DNA base-modified product generated by oxidative stress, 8 hydroxy-2'-deoxyguanosine. All three lesions are induced in a dose-dependent manner. An additional eight individuals were treated with either ultraviolet B or ultraviolet B + ultraviolet A sunblock (sun protection factor 15) and exposed to 71/2 and 15 times the minimal erythema dose on each individual, with biopsies taken at 24 h post-ultraviolet. Pyrimidine dimer and p53 expression were rarely seen in nonirradiated skin but occasional staining was seen in all normal skin for 8-hydroxy-2'-deoxyguanosine. Applications of sunscreens to human skin before irradiation were shown to attenuate erythema but did not completely eliminate all three types of cellular damage when tested up to their sun protection factor 15. Furthermore, ultraviolet B + ultraviolet A sunscreens were less efficient than the ultraviolet B alone formulation for protection against all three lesions. These results suggest that DNA damage assessed in vivo by immunohistochemistry provides a very sensitive endpoint for determining the efficacy or photosensitivity of possible different protective measures in human skin. PMID- 11886506 TI - In vivo detection of small subsurface melanomas in athymic mice using noninvasive fiber optic confocal imaging. AB - Fiber optic confocal imaging, following intravenous administration of fluorescently labeled antibodies and Texas Red-dextran, enabled in vivo detection of melanoma and surrounding blood vessels in athymic mice. Human melanoma cells (three cell lines) and cultured normal human skin cells were implanted intradermally into the haunch skin of anesthetized athymic BALB/C mice and allowed to grow to a maximum size of 2 mm diameter. Using three different fluorescein-isothiocyanate-labeled antimelanoma antibodies, single channel confocal images of melanoma cells were obtained in vivo. Using noninvasive techniques, the overall in vivo melanoma detection rate for tumors within 0.2 mm of the skin surface was 84% (27 of 32 tumors). Normal cultured human skin cells were found to have little or no fluorescence after administration of the fluorescein-isothiocyanate-labeled antibodies and tumors were not labeled by an isotype control antibody. Dual channel imaging of the implanted melanoma tumor and surrounding dermal vasculature in vivo showed increased blood vessel density at the melanoma site. Conventional immunoperoxidase histology confirmed that fiber optic confocal imaging was able to detect melanoma tumors up to 0.2 mm below the skin surface, in vivo. PMID- 11886507 TI - Fluorescence in situ detection of human cutaneous melanoma: study of diagnostic parameters of the method. AB - Multicenter study of the diagnostic parameters was conducted by three groups in Poland to determine if in situ fluorescence detection of human cutaneous melanoma based on digital imaging of spectrally resolved autofluorescence can be used as a tool for a preliminary selection of patients at increased risk of the disease. Fluorescence examinations were performed for 7228 pigmented lesions in 4079 subjects. Histopathologic examinations showed 56 cases of melanoma. A sensitivity of fluorescence detection of melanoma was 82.7% in agreement with 82.5% found in earlier work. Using as a reference only the results of histopathologic examinations obtained for 568 cases we found a specificity of 59.9% and a positive predictive value of 17.5% (melanomas versus all pigmented lesions) or 24% (melanomas versus common and dysplastic naevi). The specificity and positive predictive value found in this work are significantly lower than reported earlier but still comparable with those reported for typical screening programs. In conclusion, the fluorescence method of in situ detection of melanoma can be used in screening large populations of patients for a selection of patients who should be examined by specialists. PMID- 11886508 TI - Skin melanin, hemoglobin, and light scattering properties can be quantitatively assessed in vivo using diffuse reflectance spectroscopy. AB - Noninvasive and real-time analysis of skin properties is useful in a wide variety of applications. In particular, the quantitative assessment of skin in terms of hemoglobin and melanin content, as well as in terms of its light scattering properties, is a challenging problem in dermatology. We present here a technique for examining human skin, based on the in vivo measurement of diffuse reflectance spectra in the visible and near-infrared ranges of the electromagnetic spectrum. Spectra were measured by means of a fiber optic probe, and they were analyzed using an analytical model of light diffusion in the skin. The results of the analysis indicate that it is possible to obtain quantitative information about hemoglobin and melanin content, as well as basic information regarding the scattering properties of the skin. PMID- 11886509 TI - Degenerative alterations of dermal collagen fiber bundles in photodamaged human skin and UV-irradiated hairless mouse skin: possible effect on decreasing skin mechanical properties and appearance of wrinkles. AB - Dermal collagen fiber bundles (DCFB) are the major constructional element in the dermis. Although degenerative alterations of DCFB have been reported in chronologically aged skin, changes in photodamaged skin have not been fully investigated. We report ultrastructural alterations of DCFB, and their relation to skin elasticity using photodamaged human skin and UV-irradiated hairless mouse skin. The degree to which DCFB were intact and closely packed was evaluated and scored blindly. Exposed skin (outer forearm) exhibited marked ultrastructural degeneration. In UV-irradiated hairless mouse skin, the intact ultrastructural appearance of DCFB was gradually lost with increasing UV dosage; however, marked alterations in DCFB ultrastructure were absent in either human inner upper arm (unexposed) skin or nonirradiated age-matched control mouse skin. Skin mechanical properties were measured using a Cutometer SEM 474 suction extensometer, recording Ue* immediate deformation, Uv* viscous deformation, Uf* final deformation, and Ur* immediate contraction, all normalized for skin thickness. Uf*, Ue*, Uv*, and Ur/Uf were significantly decreased in exposed compared with unexposed skin. Significant positive correlations between degenerative alterations of DCFB and the decrease in Uf*, Ue*, and Uv* were seen. Changes of "% area of wrinkles" in UV-irradiated mouse skin was significantly correlated with degenerative changes of DCFB. Based on these results, we confirm observations made by others that chronic photodamage may have more severe effects on degeneration of DCFB than that of chronologic aging alone. Furthermore, degeneration of DCFB as detected ultrastructurally may, by its effect on skin elasticity, result in an increase in the appearance of wrinkles. PMID- 11886510 TI - Direct evidence to support the role of antigen-specific CD8(+) T cells in melanoma-associated vitiligo. AB - Vitiligo is a cutaneous pigmentary disorder characterized by the loss of melanocytes. An autoimmune mechanism is strongly suspected to be involved in this affection given that it is frequently associated with autoimmune hormonal disorders, and because antibodies directed against melanocytic antigens are found in the serum of patients with vitiligo. We examined the role of cellular immunity in melanoma-associated vitiligo by expanding infiltrating lymphocytes from fresh biopsy specimens of vitiligo patches in melanoma patients. The vitiligo infiltrating lymphocytes were almost exclusively T lymphocytes, and most were CD8(+). Following in vitro expansion, vitiligo-infiltrating lymphocytes remained predominantly CD8(+) and expressed the cutaneous homing receptor CLA. Furthermore, vitiligo-infiltrating lymphocytes had a clonal or oligoclonal T cell receptor profile, possibly reflecting specific antigenic stimulation. Finally, vitiligo- infiltrating lymphocytes specifically recognized differentiation antigens shared by normal melanocytes and melanoma cells. This direct demonstration of CD8(+) T cell involvement in vitiligo suggests that, in melanoma patients, vitiligo may be a visible effect of a spontaneous antitumoral immune response. PMID- 11886511 TI - Identification by cDNA microarray technology of genes modulated by artificial ultraviolet radiation in normal human melanocytes: relation to melanocarcinogenesis. AB - Target genes of ultraviolet stress response in cutaneous melanocytes, potentially associated with solar-induced melanocarcinogenesis, were characterized by cDNA microarray technology. In cultured normal human melanocytes, 198 genes out of approximately 9000 arrayed were found modulated > or = 1.9 times following artificial ultraviolet minus sign mainly ultraviolet-B minus sign irradiation (100 mJ per cm(2)). Among them, 159 corresponded to known sequences, the encoded proteins being mostly involved in DNA or RNA binding/synthesis/modification, or ribosomal proteins. The others were transcription factors, receptors, tumor suppressors, and (proto)oncogenes. Members of these families have already been linked to melanoma. In addition, some of the modulated genes were borne by chromosomes harboring candidate melanoma loci. Comparisons with genes modified in melanoma samples reported in previous studies with similar microarray platform showed that 59% of the known genes sensitive to ultraviolet were modulated in the same way. Furthermore, 39 expressed sequence tags were modulated, and preliminary experiments showed that two expressed sequence tags displayed differential expressions both in melanoma cell lines and in melanoma tumors. These results provide a basis for further studies on the role of modulated genes in ultraviolet induced melanoma. Because some of these genes are potential markers of the disease, they might help for developing new molecular-based strategies for risk prediction in patients. PMID- 11886512 TI - Analysis of N- and K-ras mutations in the distinctive tumor progression phases of melanoma. AB - Mutations in the ras genes are key events in the process of carcinogenesis; in particular, point mutations in codon 61 of exon 2 of the N-ras gene occur frequently in cutaneous melanoma. To investigate whether these mutations occur in early or late tumor progression phases, we searched for point mutations in the N- and K-ras genes in 69 primary cutaneous melanoma, 35 metastases, and seven nevocellular nevi in association with cutaneous melanoma. Lesions were microdissected in order to procure pure tumor samples from the distinctive growth phases of the cutaneous melanoma; the very sensitive denaturing gradient gel electrophoresis technique was used to visualize the mutations, and was followed by sequencing. Point mutations in the N-ras gene but not in the K-ras gene were detected on denaturing gradient gel electrophoresis. Twenty-three primary (33%) and nine metastatic (26%) melanomas showed bandshifts for N-ras. In the majority of cases, mutations occurring in early growth phases (i.e., the "intraepidermal" radial growth phase), were preserved in later growth phases (i.e., the invasive radial growth phase, vertical growth phase, and metastatic phase), which proves the clonal relationship between the successive growth phases. In three cases, however, the mutations differed between the distinctive growth phases within the same cutaneous melanoma, due to the occurrence of an additional mutation (especially in codon 61) in a later tumor progression phase. Our approach also permitted us to analyze the mutational status of nevi, associated with cutaneous melanoma. Six out of seven associated nevi carried the same sequence (mutated or wild-type) as the primary cutaneous melanoma, whereas in one case the sequence for N-ras differed between the primary melanoma and the associated nevus. In conclusion, this approach allowed us to demonstrate the clonal relationship between subsequent growth phases of melanoma and associated nevi; our results suggest that N-ras exon 1 mutations preferentially occur during early stages of tumor progression and hence may be involved in melanoma initiation, whereas those in N-ras exon 2 are found preferentially during later stages and hence are more probably involved in metastatic spread of cutaneous melanoma. PMID- 11886513 TI - Nucleotide excision repair genes are upregulated by low-dose artificial ultraviolet B: evidence of a photoprotective SOS response? AB - Nucleotide excision repair is a major mechanism of defense against the carcinogenic effects of ultraviolet light. Ultraviolet B causes sunburn and DNA damage in human skin. Nucleotide excision repair has been studied extensively and described in detail at the molecular level, including identification of many nucleotide excision repair-specific proteins and the genes encoding nucleotide excision repair proteins. In this study, normal human keratinocytes were exposed to increasing doses of ultraviolet B from fluorescent sunlamps, and the effect of this exposure on expression of nucleotide excision repair genes was examined. An RNase protection assay was performed to quantify transcripts from nucleotide excision repair genes, and a slot blot DNA repair activity assay was used to assess induction of the nucleotide excision repair pathway. The activity assay demonstrated that cyclobutane pyrimidine dimers were removed efficiently after exposure to low doses of ultraviolet B, but this activity was delayed significantly at higher doses. All nucleotide excision repair genes examined demonstrated a similar trend: ultraviolet B induces expression of nucleotide excision repair genes at low doses, but downregulates expression at higher doses. In addition, we show that pre-exposure of cells to low-dose ultraviolet protected keratinocytes from apoptosis following high-dose exposure. These data support the notion that nucleotide excision repair is induced in cells exposed to low doses of ultraviolet B, which may protect damaged keratinocytes from cell death; however, exposure to high doses of ultraviolet B downregulates nucleotide excision repair genes and is associated with cell death. PMID- 11886514 TI - Decreased intraindividual HLA class I expression is due to reduced transcription in advanced melanoma and does not correlate with HLA-G expression. AB - The presentation of endogenously synthesized peptides in association with HLA class I molecules allows the activation of CD8(+) lymphocytes. Tumor cells often fail to present antigenic peptides resulting in the immune escape of metastasizing cells. The aim of this study was to elucidate possible molecular mechanisms leading to reduced antigen presentation in melanoma. Melanoma cell short-time cultures were genotypically and phenotypically HLA-typed by sequence specific primer polymerase chain reaction and complement-mediated microlymphocytotoxicity assays, respectively. Flow cytometric analysis of HLA-A2 and HLA-A3 allospecificities were performed to confirm typing results. Transcriptional levels of classical HLA-A, HLA-B genes and nonclassical HLA-G genes were detected using quantitative real-time reverse transcriptase polymerase chain reaction (LightCycler). We found loss or downregulation of HLA proteins in 18% (for HLA-A) and 53% (for HLA-B) of all tested metastases. Genomic analysis, however, revealed the presence of the corresponding HLA class I gene in six out of seven cases. On the level of gene transcription we observed a differential regulation of HLA-A, HLA-B, and HLA-G mRNA expression. There was no correlation between classical and nonclassical HLA gene transcription, but the transcriptional levels of classical HLA corresponded to the protein expression levels. Furthermore, an overall reduced amount of HLA class I gene transcription was observed in melanoma metastases during disease progression in three individuals. We postulate that there is a transcriptional regulation of HLA class I gene expression in melanoma cells. These data suggest that treatment approaches aimed at activating specific cytotoxic T lymphocytes are most successful in early disease. PMID- 11886515 TI - Transcriptional repression of the microphthalmia gene in melanoma cells correlates with the unresponsiveness of target genes to ectopic microphthalmia associated transcription factor. AB - In the melanocyte, expression of genes required for pigment formation is mediated by the microphthalmia transcription factor, which is also critical for the development and survival of normal melanocytes during embryogenesis. Here we show that the expression of the melanocyte-specific isoform of microphthalmia transcription factor is lost in a subset of human melanoma cell lines, accompanied by the repression of tyrosinase and tyrosinase-related proteins 1 and 2, the three transcriptional target genes for microphthalmia. After the forced expression of microphthalmia transcription factor in melanoma cells where the expression of endogenous microphthalmia gene was found to be extinguished, no restoration of the melanogenic phenotype occurred and the transcription of the three microphthalmia transcription factor target genes remained silent. The transcription activation domain of microphthalmia transcription factor, tested as a GAL-MITF fusion protein, remained fully functional in these cells, however, and ectopic microphthalmia transcription factor localized normally to the nucleus and bound to the tyrosinase initiator E-box in gel retardation assays. Thus, the block of differentiation in microphthalmia-transcription-factor-negative melanomas extended the transcriptional repression of the microphthalmia transcription factor gene alone, and endogenous promoters in these melanoma cells became no longer responsive to microphthalmia transcription factor when this was substituted exogenously. The data presented suggest that a specific nuclear context is required for the transcriptional activation of the melanocyte markers by the microphthalmia transcription factor in malignant melanocytes and this specificity is lost concomitantly with the transcriptional repression of microphthalmia transcription factor. PMID- 11886516 TI - Primary cutaneous follicle center cell lymphomas and large B cell lymphomas of the leg descend from germinal center cells. A single cell polymerase chain reaction analysis. AB - Primary cutaneous B cell lymphomas are defined as non-Hodgkin lymphomas that occur in the skin without extracutaneous involvement for 6 mo after diagnosis. They are characterized by a less aggressive course and better prognosis than their nodal counterparts. According to the European Organization for Research and Treatment of Cancer classification, the major subentities of primary cutaneous B cell lymphoma are follicle center cell lymphomas, immunocytomas, and large B cell lymphomas of the leg, which differ considerably regarding their clinical behavior, the former two being indolent, the latter being of intermediate malignancy. In this study, we applied a single cell polymerase chain reaction approach to analyze immunoglobulin V(H)/V(L) genes in 532 individual B lymphocytes from histologic sections of four follicle center cell lymphomas localized on the head and trunk, and four large B cell lymphomas on the leg. We found: (i) in six of eight patients a clonal heavy chain, and in seven of eight patients a clonal light chain rearrangement, all being potentially productive; (ii) no bias in VH gene usage, in four of seven light chain rearrangements the V kappa germline gene IGVK3-20*1 was used; (iii) no biallelic rearrangements; (iv) all V(H)/V(L) genes are extensively mutated (mutation rate 5.4-16.3%); (v) intraclonal diversity in six of eight cases (three of each group); and (vi) low replacement vs silent mutation ratios in framework regions indicating preservation of antigen-receptor structure, as in normal B cells selected for antibody expression. Our data indicate a germinal center cell origin of primary cutaneous follicle center cell lymphomas and large B cell lymphomas independent of those belonging to one of these subentities. PMID- 11886517 TI - Haplotype analysis in determination of the heredity of erythropoietic protoporphyria among Swiss families. AB - Defects in the human ferrochelatase gene lead to the hereditary disorder of erythropoietic protoporphyria. The clinical expression of this autosomal dominant disorder requires an allelic combination of a disabled mutant allele and a low expressed nonmutant allele. Unlike most other erythropoietic protoporphyria populations, mutations identified among Swiss erythropoietic protoporphyria families to date have been relatively homogeneous. In this study, genotype analysis was conducted in seven Swiss erythropoietic protoporphyria families, three carrying mutation Q59X, two carrying mutation insT213, and two carrying mutation delTACAG(580-584). Three different haplotypes of five known intragenic single nucleotide polymorphisms, namely -251 A/G, IVS1-23C/T, 798 G/C, 921 A/G, and 1520C/T, were identified. Each haplotype was shared by families carrying an identical mutation in the ferrochelatase gene indicating a single mutation event for each of the three mutations. These mutations have been present in the Swiss erythropoietic protoporphyria population for a relatively long time as no common haplotypes of microsatellite markers flanking the ferrochelatase gene were found, except of two conserved regions, telomeric of the insT213 allele and centromeric of the delTACAG(580-584)allele, each with a size > 3 cM. Among the nonmutant ferrochelatase alleles, patients from six erythropoietic protoporphyria families shared a common haplotype [-251G; IVS1-23T] of the first two single nucleotide polymorphisms. An exception was the haplotype [-251 A; IVS1-23C] identified in the index patient of one erythropoietic protoporphyria family. These results supported the recent findings that the low expressed allele is tightly linked to a haplotype [-251G; IVS1-23T] of two intragenic single nucleotide polymorphisms in the ferrochelatase gene. PMID- 11886518 TI - An application of the United Kingdom Working Party diagnostic criteria for atopic dermatitis in Scottish infants. AB - The United Kingdom Working Party diagnostic criteria for atopic dermatitis have been characterized in infants and children; however, the need for visual confirmation of flexural dermatitis by a trained investigator limits their use in large epidemiologic studies. We have administered the complete United Kingdom Working Party criteria in a postal questionnaire format to the mothers of year old infants and determined the concordance between mothers' and trained investigator's reports of visual flexural dermatitis. Based on mothers' responses to the questionnaire, 59 infants with atopic dermatitis and 59 controls were identified. In subsequent home interviews conducted by a trained investigator, the United Kingdom criteria questions were repeated and sites of current visible dermatitis were identified by mothers and the investigator as per United Kingdom Working Party protocol. Agreement between the mothers' postal and home interview responses was high: kappa= 0.75-0.94 for individual criteria; kappa= 0.93 for diagnosed atopic dermatitis. Agreement between the mothers' and investigator's observations of visible flexural dermatitis was high for all sites: kappa= 0.88 1.0. The results demonstrate that mothers are able to apply the United Kingdom criteria and accurately report visible flexural dermatitis in their year old infants. The postal application of the United Kingdom Working Party's diagnostic criteria for atopic dermatitis in year old infants appears to be a practical, reliable, epidemiologic tool in the investigation of atopic dermatitis with results comparable with formal application of the criteria by a trained investigator. PMID- 11886519 TI - Cancer risk in a population-based cohort of patients hospitalized for psoriasis in Sweden. AB - Studies of clinical series of psoriasis patients have suggested an increased risk of nonmelanoma skin cancer and melanoma; the risk of other neoplasms has rarely been studied. In order to assess the incidence of cancer in a nationwide series of psoriasis patients from Sweden, we followed up, for the years 1965-89, 9773 patients with a hospital discharge diagnosis of psoriasis made during 1965-83, who were alive and free from malignancy 1 y after first discharge. We compared their incidence of neoplasms with that of the national population by computing standardized incidence ratios (SIR). We observed a total of 789 neoplasms [SIR 1.37, 95% confidence interval (CI) 1.28, 1.47]. There was an increase in the risk of cancers of the oral cavity and pharynx (SIR 2.80, 95% CI 1.96, 3.87), liver (SIR 1.91, 95% CI 1.28, 2.74), pancreas (SIR 1.56, 95% CI 1.02, 2.23), lung (SIR 2.13, 95% CI 1.71, 2.61), skin (squamous cell carcinoma, SIR 2.46, 95% CI 1.82, 3.27), female breast (SIR 1.27, 95% CI 1.00, 1.58), vulva (SIR 3.24, 95% CI 1.18, 7.06), penis (SIR 4.66, 95% CI 1.50, 10.9), bladder (SIR 1.43, 95% CI 1.03, 1.92), and kidney (SIR 1.56, 95% CI 1.04, 2.25). The risk of malignant melanoma was decreased (SIR 0.32, 95% CI 0.10, 0.74). Despite some limitations (possible diagnostic misclassification, lack of data on treatment, relatively short follow up), our study provides evidence against an increased risk of melanoma among patients hospitalized for psoriasis. In addition to nonmelanoma skin and genital cancers, patients hospitalized for psoriasis were at increased risk of several malignancies, in particular those associated with alcohol drinking and tobacco smoking. PMID- 11886520 TI - Basal keratinocytes from uninvolved psoriatic skin exhibit accelerated spreading and focal adhesion kinase responsiveness to fibronectin. AB - We previously proposed that the keratinocyte hyperproliferative state in psoriatic skin results from a combination of T cell cytokine interaction with basal keratinocytes that exist in a primed state. We now provide evidence that basal keratinocytes from psoriatic uninvolved skin are in a preactivated state with regard to their interaction with fibronectin. Freshly isolated basal keratinocytes (K(1)/K(10)(-)) from non-lesional psoriatic skin demonstrated a significantly higher percentage of spreading cells 1 h after plating on fibronectin-coated plates than keratinocytes isolated from normal skin (p =0.0002). No differences were observed on collagen-laminin-coated plates, however. The keratinocyte spreading on fibronectin-coated plates involved alpha 5 beta 1 and alpha V beta 1 integrins. To address the potential signaling cascades that may respond to integrin changes in psoriatic keratinocytes, focal adhesion kinase changes were assessed. The percentage of keratinocytes from psoriatic uninvolved skin that exhibit positive focal adhesion kinase staining was significantly greater than the percentage from healthy volunteers after 1 h incubation on fibronectin (p =0.006). Additionally, focal adhesion kinase isolated from uninvolved psoriatic keratinocytes had a greater degree of tyrosine phosphorylation. Thus, the proliferative effect of fibronectin in combination with T cell lymphokines on psoriatic uninvolved basal keratinocyte progenitors may be due to abnormal in vivo integrin-driven focal adhesion kinase activity and downstream signaling. PMID- 11886521 TI - Oligoclonal expansion of intraepidermal T cells in psoriasis skin lesions. AB - CD8(+) T cell infiltration into the epidermis is thought to be a key event in the pathogenesis of psoriasis. A quantitative competitive polymerase chain reaction method was developed to examine the expression of T cell receptor beta chain variable region 2, 3, 6.1-3, 8, and 13.1 genes in the epidermis of psoriatic lesions. Paired epidermal samples and peripheral blood samples from five psoriasis patients were studied. The results demonstrated the expansion of T cell receptor beta chain variable region 3 (two patients), 8 (two patients), and/or 2 (one patient). Contrary to previous reports, neither beta chain variable region 6.1-3 nor beta chain variable region 13.1 subgroups were expanded in any of the lesions. DNA sequence analysis revealed dominant T cell clones observed in all expanded beta chain variable region families and heterogeneous populations and/or small clones observed in non-expanded beta chain variable region families. Using CDR3 length analysis to examine the complete beta chain repertoire of the infiltrating T cells in the lesional epidermis, we found that approximately 50% of the T cell receptor beta chain variable region families in each patient's lesion demonstrated abnormal CDR3 DNA length distribution, indicating the presence of monoclonal or oligoclonal T cell expansion. Together, the results show that among different patients, T cell oligoclonality is not restricted to a limited number of T cell receptor beta chain variable region families. In an attempt to identify the pathogenic T cells among the many expanded T cell clones in the lesions, we compared T cell receptor expansion in the lesional epidermis with non-lesional epidermis. Particular T cell receptor were found to be preferentially expanded in lesional epidermis and these lesion-specific T cell clones may be most important in the pathogenesis and development of psoriatic lesions. PMID- 11886522 TI - Osteopontin gene is expressed in the dermal papilla of pelage follicles in a hair cycle-dependent manner. AB - Hair follicle formation and maintenance involve intimate interactions between follicular epithelial cells and a group of specialized mesenchymal cells known as the dermal papilla. Using the random primer polymerase chain reaction, we have identified an approximately 1.4 kb osteopontin mRNA that is present in large quantities in cultured rat vibrissa dermal papilla cells but undetectable in cultured rat skin fibroblasts. In situ hybridization showed that the osteopontin gene is expressed in dermal papilla cells of pelage follicles during catagen but not in anagen or telogen. As an acidic glycosylated RGD-containing extracellular matrix protein, osteopontin can function both as a cell attachment protein and as a soluble cytokine playing roles in signaling, cell migration, tissue survival, anti-inflammation, and T-cell-mediated cellular immunity. Our results indicate that the comparison of the mRNA of cultured dermal papilla cells and fibroblasts can lead to the identification of not only anagen-specific genes (e.g., nexin 1), but also a catagen-specific gene. We have thus provided evidence that specific genes are turned on during catagen, which is therefore not simply a passive "degenerative" phase. The functional role of osteopontin in catagen is unclear but it may promote the formation of a tightly aggregated dermal papilla, and/or protect the dermal papilla cells from apoptosis induced by cytokines or hypoxia during catagen. PMID- 11886523 TI - Transcriptional regulators of steroidogenesis, DAX-1 and SF-1, are expressed in human skin. AB - DAX-1 and SF-1 are members of the orphan nuclear receptor superfamily that are critical regulatory components of the hypothalamic-pituitary-adrenal-gonadal axis. In adrenal and gonadal tissues they regulate the expression of the cytochrome P450 steroid hydroxylase genes, key mediators of steroidogenesis. The identification of a number of steroid hydroxylases in human skin prompted us to investigate the presence of DAX-1 and SF-1. Immuno histochemical analysis of human skin revealed a distinctive staining pattern for DAX-1 and SF-1 in skin and its appendages. Prominent staining for DAX-1 was confined to the epidermis, sebaceous glands, sweat glands, and outer root sheath of the hair follicle with weaker expression in the inner root sheath, matrix cells, and dermal papilla cells. Similarly, SF-1 was also detected in the epidermis but displayed a scattered nuclear pattern across all layers. SF-1 immunoreactivity was also detected in the exocrine glands and was stronger than DAX-1 in the inner root sheath, matrix cells, and dermal papilla cells. Co-localization of DAX-1 and SF-1 was demonstrated by immunocytochemistry in the HaCaT keratinocyte cell line, primary keratinocytes, preadipocytes, and dermal papilla cells. Reverse transcriptase-polymerase chain reaction analysis demonstrated the expression of DAX-1 and SF-1 mRNA in whole human skin and Western analysis also confirmed the presence of DAX-1 protein in skin-derived cells. Our investigations demonstrate that two important regulators of steroidogeneisis are present in human skin and its appendages. These transcription factors may have a role in cutaneous steroidogenesis and thus be involved in hair follicle cycling or pathologies associated with steroids. Further studies are needed to determine the functional roles of DAX-1 and SF-1 in human skin. PMID- 11886524 TI - PARP determines the mode of cell death in skin fibroblasts, but not keratinocytes, exposed to sulfur mustard. AB - Sulfur mustard is cytotoxic to dermal fibroblasts as well as epidermal keratinocytes. We demonstrated that poly(ADP-ribose) polymerase (PARP) modulates Fas-mediated apoptosis, and other groups and we have shown that PARP plays a role in the modulation of other types of apoptotic and necrotic cell death. We have now utilized primary dermal fibroblasts, immortalized fibroblasts, and keratinocytes derived from PARP(-/-) mice and their wildtype littermates (PARP(+/+)) to determine the contribution of PARP to sulfur mustard toxicity. Following sulfur mustard exposure, primary skin fibroblasts from PARP-deficient mice demonstrated increased internucleosomal DNA cleavage, caspase-3 processing and activity, and annexin V positivity, compared to those derived from PARP(+/+) animals. Conversely, propidium iodide staining, PARP cleavage patterns, and random DNA fragmentation revealed a dose-dependent increase in necrosis in PARP(+/+) but not PARP(-/-) cells. Using immortalized PARP(-/-) fibroblasts stably transfected with the human PARP cDNA or with empty vector alone, we show that PARP inhibits markers of apoptosis in these cells as well. Finally, primary keratinocytes were derived from newborn PARP(+/+) and PARP(-/-) mice and immortalized with the E6 and E7 genes of human papilloma virus. In contrast to fibroblasts, keratinocytes from both PARP(-/-) and PARP(+/+) mice express markers of apoptosis in response to sulfur mustard exposure. The effects of PARP on the mode of cell death in different skin cell types may determine the severity of vesication in vivo, and thus have implications for the design of PARP inhibitors to reduce sulfur mustard pathology. PMID- 11886525 TI - Appearance of Langerhans cells in the epidermis of Tgfb1(-/-) SCID mice: paracrine and autocrine effects of transforming growth factor-beta 1 and -beta 2(1). AB - A striking immunologic abnormality of normal and SCID Tgfb1(-/-) mice is the total absence of Langerhans cells in their epidermis. Here we show that transfer of Tgfb1(+/-) SCID bone marrow causes, within a few weeks, the appearance of Langerhans cells in the epidermis of gamma-irradiated and unirradiated Tgfb1(-/-) SCID recipients. In addition, local injection of 2 x 10(5) latent transforming growth factor-beta1 cDNA-transduced cloned CD4+ T lymphocytes causes the appearance of Langerhans cells in the ear epidermis of Tgfb1(-/-) SCID mice. This effect is enhanced by antigen-specific activation of these T cells. Injection of recombinant active transforming growth factor-beta 2 into the ear of Tgfb1(-/-) SCID mice also results in the migration of Langerhans cells into the epidermis locally, but no epidermal Langerhans cells are seen after systemic injections of transforming growth factor-beta 2. Our results suggest that transforming growth factor-beta can act in paracrine as well as autocrine fashion to induce the differentiation of precursors into Langerhans cells. Furthermore, these results indicate that the relative roles of different transforming growth factor-beta isoforms in vivo may be influenced by their local availability and/or the regulation of their conversion from latent into active form. PMID- 11886526 TI - Decreased mRNA stability as a mechanism of glucocorticoid-mediated inhibition of vascular endothelial growth factor gene expression by cultured keratinocytes. AB - Epidermal keratinocyte-derived overexpression of vascular endothelial growth factor has been functionally linked to increased density of tortuous and hyperpermeable dermal microvessels, representing a characteristic component of cutaneous inflammation. We hypothesized that potent anti-inflammatory properties of synthetic glucocorticoids are attributed in part to their interference with the regulated vascular endothelial growth factor expression by keratinocytes. As vascular endothelial growth factor is markedly upregulated by autocrine transforming growth factor alpha and paracrine hepatocyte growth factor/scatter factor expression, the effect of glucocorticoids on growth-factor-induced vascular endothelial growth factor production by primary and immortalized keratinocytes was examined. Glucocorticoids were shown to suppress vascular endothelial growth factor protein and mRNA expression in a concentration- and time-dependent fashion. In transcriptional activation studies, however, common 5' regulatory regions of the vascular endothelial growth factor gene failed to confer inhibitory glucocorticoid effects. Instead, glucocorticoids were shown to increase vascular endothelial growth factor mRNA turnover, indicating that post transcriptional modes of glucocorticoid action are employed to negatively regulate induced vascular endothelial growth factor expression. Together, these studies identify vascular endothelial growth factor upregulation by epidermal keratinocytes as a putative target of glucocorticoid action in cutaneous inflammation. Our data provide strong evidence that mRNA destabilization may represent a mechanism by which glucocorticoids inhibit growth-factor-regulated vascular endothelial growth factor gene expression by keratinocytes. PMID- 11886527 TI - 8-Cl-adenosine induces growth arrest without differentiation of primary mouse epidermal keratinocytes. AB - In some cell systems, the antiproliferative effects of 8-Cl-cAMP, a site selective cAMP analog specific for the type I cAMP-dependent protein kinase, are mediated by its metabolite, 8-Cl-adenosine. These effects were once thought to be specific to transformed cells. We investigated the ability of 8-Cl-adenosine to regulate growth and differentiation in primary cultures of mouse epidermal keratinocytes. A 24 h exposure of keratinocytes to 8-Cl-adenosine inhibited [3H]thymidine incorporation in a dose-dependent manner with an apparent IC(50) of 7.5 microM, and these effects were completely reversible. To determine the ability of 8-Cl-adenosine to induce differentiation of primary keratinocytes, we measured keratin-1 expression and transglutaminase activity, markers of early and later stages of keratinocyte differentiation, respectively. Interestingly, exposure of keratinocytes to 25 microM 8-Cl-adenosine for 24 h had no effect on keratin-1 expression or transglutaminase activity. The 8-Cl-adenosine-induced growth arrest of keratinocytes required uptake of the compound and was accompanied by an increase in protein expression of the cyclin-dependent protein kinase inhibitor p21(WAF1/Cip1). These results demonstrate that 8-Cl-adenosine inhibits growth in a non-transformed/non-immortalized cell system, possibly through an elevation in p21(WAF1/Cip1) protein levels, without inducing differentiation. PMID- 11886528 TI - Minoxidil-induced hair growth is mediated by adenosine in cultured dermal papilla cells: possible involvement of sulfonylurea receptor 2B as a target of minoxidil. AB - The mechanism by which minoxidil, an adenosine-triphosphate-sensitive potassium channel opener, induces hypertrichosis remains to be elucidated. Minoxidil has been reported to stimulate the production of vascular endothelial growth factor, a possible promoter of hair growth, in cultured dermal papilla cells. The mechanism of production of vascular endothelial growth factor remains unclear, however. We hypothesize that adenosine serves as a mediator of vascular endothelial growth factor production. Minoxidil-induced increases in levels of intracellular Ca(2+) and vascular endothelial growth factor production in cultured dermal papilla cells were found to be inhibited by 8-sulfophenyl theophylline, a specific antagonist for adenosine receptors, suggesting that dermal papilla cells possess adenosine receptors and sulfonylurea receptors, the latter of which is a well-known target receptor for adenosine-triphosphate sensitive potassium channel openers. The expression of sulfonylurea receptor 2B and of the adenosine A1, A2A, and A2B receptors was detected in dermal papilla cells by means of reverse transcription polymerase chain reaction analysis. In order to determine which of the adenosine receptor subtypes contribute to minoxidil-induced hair growth, the effects of subtype-specific antagonists for adenosine receptors were investigated. Significant inhibition in increase in intracellular calcium level by minoxidil or adenosine was observed as the result of pretreatment with 8-cyclopentyl-1,3-dipropylxanthine, an antagonist for adenosine A1 receptor, but not by 3,7-dimethyl-1-propargyl-xanthine, an antagonist for adenosine A2 receptor, whereas vascular endothelial growth factor production was blocked by both adenosine A1 and A2 receptor antagonists. These results indicate that the effect of minoxidil is mediated by adenosine, which triggers intracellular signal transduction via both adenosine A1 and A2 receptors, and that the expression of sulfonylurea receptor 2B in dermal papilla cells might play a role in the production of adenosine. PMID- 11886529 TI - The p38-MAPK/SAPK pathway is required for human keratinocyte migration on dermal collagen. AB - Human keratinocyte motility plays an important role in the re-epithelialization of human skin wounds. The wound bed over which human keratinocytes migrate is rich in extracellular matrices, such as fibrin, fibronectin, and collagen, and serum factors, such as platelet-derived growth factor and transforming growth factor beta 1. Extracellular matrices and the serum factors bind to cell surface receptors and initiate a cascade of intracellular signaling events that regulate cell migration. In this study, we identified an intracellular signaling pathway that mediates collagen- driven motility of human keratinocytes. Pharmaco logic inhibition of the activation of p38-alpha and p38-beta mitogen-activated protein kinase activation potently blocked collagen-driven human keratinocyte migration. Transfection of the same keratinocytes with the kinase-negative mutants of p38 alpha or p38-beta mitogen-activated protein kinase markedly inhibited keratinocyte migration on collagen. Attachment of keratinocytes to collagen activated p38 mitogen- activated protein kinase, as well as p44/p42 ERKs. Interestingly, activation of the p38 mitogen-activated protein kinase cascade by overexpressing the constitutively active MKK3 and MKK6, MKK3b(E) and MKK6b(E), could neither initiate migration in the absence of collagen nor enhance collagen driven migration. This study provides evidence that the p38-MAPK/SAPK pathway is necessary, but insufficient, for mediating human keratinocyte migration on collagen. PMID- 11886530 TI - Assembly of epithelial cell fibrillins. AB - Fibrillins are large structural macromolecules that are components of connective tissue microfibrils. Fibrillin microfibrils have been found in association with basement membranes, where microfibrils appear to insert directly into the lamina densa. It is unknown whether fibrillins are limited to these sites of microfibril insertion or are present throughout the lamina densa. In this study, electron microscopic immunolocalization demonstrated the presence of fibrillin-1 throughout the lamina densa in the dermal-- epidermal junction. In order to investigate whether fibrillin microfibrils might be present in the lamina densa, epithelial cell cultures (WISH, HaCaT, and primary keratinocytes) were analyzed by immunofluorescence, immunoblotting, and extraction of microfibrils followed by rotary shadowing electron microscopy and compared to mesenchymal cell cultures (dermal fibroblasts and MG63 osteosarcoma). In contrast to mesenchymal cells, which elaborate a fibrillin fibril network, epithelial cells primarily deposit fibrillin into the extracellular matrix in a nonfibrillar form. Coculture experiments using human epithelial cells and mouse fibroblasts implicated the cells themselves in the assembly of fibrillin. The importance of the cell in this process was further underscored by novel data demonstrating that keratinocytes selectively secrete fibrillin-1 into the matrix and not into the medium and can differentiate between fibrillin-1 and fibrillin-2. PMID- 11886531 TI - Involvement of leukotriene B(4) in substance P-induced itch-associated response in mice. AB - Intradermal injection of substance P elicits an itch sensation in human subjects and an itch-associated response in mice. The substance P-induced itch-associated response in mice is not inhibited by antihistamine. Therefore, the mechanisms of substance P-induced itch-associated response are unclear. In this study, we demonstrated one of the mechanisms. Substance P induces an arachidonate cascade to produce prostaglandins and leukotriene. In this study we considered whether arachidonate metabolites are involved in the substance P-induced itch-associated response. A phospholipase A(2) inhibitor arachidoryltrifluoromethyl ketone inhibited the substance P-induced itch-associated response in mice. Pre treatment with the glucocorticoids betamethasone and dexamethasone also produced inhibition of the substance P-induced itch-associated response in mice as well as humans. The 5-lipoxygenase inhibitor zileuton, but not the cyclooxygenase inhibitors indomethacin and diclofenac, suppressed substance P-induced itch-associated response. The leukotriene B(4) receptor antagonist 5-[2-(2-carboxyethyl)-3-[6-(4 methoxyphenyl)-5E-hexenyl]oxyphenoxy]valeric acid produced inhibition, whereas pranlukast (leukotriene C(4)/D(4)/E(4) receptor antagonist) and 5(Z)-7-[1S,2S, 3S,5R-3-(trans-b-styren)sulfonamido-6,6-dimethylbi cyclo(3,1,1)hept-2-yl]-5 heptenoic acid (EP(1) receptor antagonist) were without effect. Furthermore, when the production of leukotriene B(4) and prostaglandin E(2) was measured in skin injected with substance P and in mouse keratinocytes applied with substance P, the level of both products increased. As leukotriene B(4), but not prostaglandin E(2), also induces the itch-associated response in mice, these results suggest that leukotriene B(4) and keratinocytes, cutaneous cells which produced leukotriene B(4), play an important role in substance P-induced itch-scratch response in mice. Leukotriene B(4) receptor antagonist and 5-lipoxygenase inhibitor may be novel antipruritic drugs. PMID- 11886532 TI - Cyclosporin A exacerbates skin irritation induced by tributyltin by increasing nuclear factor kappa B activation. AB - In searching for pharmacologic agents able to reduce xenobiotic-induced skin irritation, we found that cyclosporine A exacerbates the skin irritation induced by tributyltin. We previously demonstrated the involvement of interleukin-1 alpha and tumor necrosis factor alpha in tributyltin-induced skin irritation. Here, we show that cyclosporine A (28 mg per kg), at a dose that results in systemic immunosuppression, potentiates tributyltin-induced skin irritation through increased tumor necrosis factor alpha production, associated with increased tributyltin-induced activation of transcription factor nuclear factor kappa B in cyclosporine-A-treated mice. On the other hand, under the same experimental conditions, cyclosporine A prevented the elicitation phase of oxazolone-induced contact allergy, but was ineffective in preventing benzalkonium-chloride-induced skin irritation. Using a murine keratinocyte cell line (HEL30) we demonstrated, also in vitro, that the cyclosporine A potentiates tributyltin-induced nuclear factor kappa B activation and cytokine production, this being preceded by an increase in cellular oxidative activity, essential for nuclear factor kappa B activation, that is time and dose (0.1-10 microM) dependent. This effect was not exclusive to tributyltin but could be extended to other mitochondrial poisons such as sodium arsenate. It has been reported that cyclosporine A binds to cyclophilins. An 18-mer antisense phosphorothioate oligodeoxynucleotide was used to target mitochondrial cyclophilin D mRNA. After 24 h exposure to the oligonucleotide, the amount of cyclophilin D in the cells was decreased by 54% as judged by Western blot analysis. Cyclophilin D suppression prevented cyclosporine A potentiation of tributyltin-induced cellular oxidative activity, indicating the key role of the binding of cyclosporine A to mitochondrial cyclophilin D in mediating this effect. PMID- 11886533 TI - Anti-mycotics suppress interleukin-4 and interleukin-5 production in anti-CD3 plus anti-CD28-stimulated T cells from patients with atopic dermatitis. AB - It is reported that anti-mycotic agents are effective for the treatment of patients with atopic dermatitis. We studied the in vitro effects of anti-mycotics on T helper-1 and T helper-2 cytokine production in anti-CD3 plus anti-CD28 stimulated T cells from atopic dermatitis patients and normal donors. The amounts of interleukin-4 and interleukin-5 secreted by anti-CD3/CD28-stimulated T cells were higher in atopic dermatitis patients than in normal donors. Azole derivatives, ketoconazole, itraconazole, miconazole, and nonazole terbinafine hydrochloride, and tolnaftate reduced interleukin-4 and interleukin-5 secretion without altering that of interferon-gamma and interleukin-2 in anti-CD3/CD28 stimulated T cells from both atopic dermatitis patients and normal donors. The azole derivatives were more inhibitory than nonazole anti-mycotics. These anti mycotics reduced the anti-CD3/CD28-induced mRNA expression and promoter activities for interleukin-4 and interleukin-5. The 3',5'-cyclic adenosine monophosphate analog dibutyryl 3',5'-cyclic adenosine monophosphate reversed the inhibitory effects of the anti-mycotics on interleukin-4 and interleukin-5 secretion, mRNA expression, and promoter activities. Anti-CD3/CD28 transiently (< or = 5 min) increased intracellular 3',5'-cyclic adenosine monophosphate in T cells, and the increase was greater in atopic dermatitis patients than in normal donors. The increase of 3',5'-cyclic adenosine monophosphate by anti-CD3/CD28 correlated with interleukin-4 and interleukin-5 secretion by anti-CD3/CD28. The transient 3',5'-cyclic adenosine monophosphate increase was suppressed by anti mycotics, and azole derivatives were more suppressive than nonazoles. Azole derivatives inhibited the activity of cyclic adenosine monophosphate-synthesizing adenylate cyclase whereas terbinafine hydrochloride and tolnaftate enhanced the activity of 3',5'-cyclic adenosine monophosphate-hydrolyzing cyclic nucleotide phosphodiesterase in atopic dermatitis and normal T cells. These results suggest that the anti-mycotics may suppress interleukin-4 and interleukin-5 production by reducing 3',5'-cyclic adenosine monophosphate signal, and stress their potential use for the suppression of T helper-2-mediated allergic reactions. PMID- 11886534 TI - Late-onset erythropoietic porphyria caused by a chromosome 18q deletion in erythroid cells. AB - The erythropoietic porphyrias, erythropoietic protoporphyria and congenital erythropoietic porphyria, result from germline mutations in the ferrochelatase gene and uroporphyrinogen III synthase gene, respectively. Both conditions normally present in childhood but rare cases with onset past the age of 40 y have been reported. Here we show that late-onset erythropoietic protoporphyria can be caused by deletion of the ferrochelatase gene in hematopoietic cells with clonal expansion as part of the myelodysplastic process. This is the first direct demonstration of porphyria produced by an acquired molecular defect restricted to one tissue. Some other cases of late-onset erythropoietic porphyria may be explained by a similar mechanism. PMID- 11886535 TI - Activation of a cryptic splice site of PTEN and loss of heterozygosity in benign skin lesions in Cowden disease. AB - Cowden disease is an autosomal dominant syndrome characterized by facial trichilemmomas, acral keratoses, papillomatous papules, mucosal lesions, and an increased risk for breast and nonmedullary thyroid cancer. Here, we describe a novel PTEN splicing site mutation in a family with classical Cowden disease and we studied benign skin lesions typical for Cowden disease for loss of heterozygosity. We found a PTEN IVS2 + 1G > Alpha 5'-splicing acceptor mutation resulting in activation of a cryptic splice site. Activation of this cryptic splice site is predicted to result in a frameshift with a premature stop codon, thus disrupting the phosphatase core motif of PTEN. Loss of heterozygosity analysis of two trichilemmomas, one fibroma, and three acanthomas of the index patient demonstrated loss of heterozygosity at the PTEN locus in four of these lesions. In conclusion, our data demonstrate that a PTEN splicing site mutation causes activation of a cryptic splice site, which results in aberrant transcripts. PMID- 11886536 TI - Mutations of ATP2C1 in Japanese patients with Hailey-Hailey disease: intrafamilial and interfamilial phenotype variations and lack of correlation with mutation patterns. AB - We report herein mutations of ATP2C1 in 11 Japanese patients with Hailey-Hailey disease gene (including five previously reported) and compare the mutation pattern with clinical phenotypes. Patients with missense mutations and some of those with mutations causing premature termination showed erythema and erosions primarily at intertriginous areas. In two families with unique mutations, one with an in-frame three amino acid deletion plus an eight amino acid insertion and one with a two base pair deletion predicted to cause premature truncation, some affected individuals had unique clinical features -- generalization of Hailey Hailey disease and generalized skin eruption resembling keratotic papules in Darier's disease -- but other affected individuals did not, suggesting the presence of severe intrafamilial phenotype variations. Our findings suggest that differences in clinical phenotypes are probably related to factors other than the type of causative mutation. PMID- 11886537 TI - Novel point mutations, deletions, and polymorphisms in the cathepsin C gene in nine families from Europe and North Africa with Papillon-Lefevre syndrome. AB - Papillon-Lefevre syndrome is an autosomal recessive disorder characterized by palmoplantar keratoderma, periodontitis, and premature loss of dentition. Mutations in the CTSC gene that encodes cathepsin C have been described in families affected with Papillon--Lefevre syndrome. Cathepsin C is the least understood of the lysosomal cysteine proteases; it has been reported to participate in both intracellular and extracellular cleavage of proteins and activation of serine proteases in immune and inflammatory cells. We report here eight new mutations in Papillon-Lefevre syndrome families: four deletions and four point mutations, including a missense mutation in the propeptide chain that could help elucidate structure-function relationships in this protein. We also found that the 458C > T mutation, first reported in two families by Hart et al (2000c), was a neutral polymorphism in our families, as suggested by Allende et al (Cathepsin C gene: first compound heterozygous patient with Papillon--Lefevre syndrome and novel symptomless mutation. Hum Mutat 17:152-153, 2001). PMID- 11886538 TI - Clinical and molecular diagnostic criteria of congenital atrichia with papular lesions. AB - Congenital atrichia with papular lesions is a rare, autosomal recessive form of total alopecia and mutations in the hairless (hir) gene have been implicated in this disorder. Published estimates of the prevalence of this disorder remain surprisingly low considering pathogenetic mutations in hir have been found in distinct ethnicities around the world. Therefore, it is likely that congenital atrichia with papular lesions is far more common than previously thought and is often mistaken for its phenocopy, the putative autoimmune form of alopecia universalis. To clarify this discrepancy, we propose criteria for the clinical diagnosis of congenital atrichia with papular lesions. Among these is the novel report of the consistent observation of hypopigmented whitish streaks on the scalp surface of affected individuals. Additionally, we report the identification of a novel missense mutation in hir from a family of Arab Palestinian origin that exhibits the pathognomonic features of atrichia with papular lesions. Collectively, we anticipate that an increased recognition of this disorder will result in more accurate diagnosis and the sparing of unnecessarily treatment to patients. PMID- 11886539 TI - No evidence of deregulated patched-hedgehog signaling pathway in trichoblastomas and other tumors arising within nevus sebaceous. AB - Nevus sebaceous is a congenital malformation of the skin within which a number of neoplasms showing adnexal differentiation may arise. Recently, deletions in the patched gene region were reported in nevus sebaceous and constitutive activation of the patched-hedgehog signaling pathway was implicated in the development of tumors arising within nevus sebaceous. To substantiate further a role of the patched-hedgehog signaling pathway in secondary tumors arising within nevus sebaceous, we examined 11 nevus sebaceous associated with secondary tumors for loss of heterozygosity of the patched gene region by microsatellite polymerase chain reaction and patched mRNA expression by in situ hybridization. Unexpectedly, however, none of the tumors (including eight trichoblastomas) and nevus sebaceous lesions showed loss of heterozygosity at any polymorphic loci close to the patched gene. Further more, none of the nevus sebaceous lesions and secondary tumors gave detectable signals for patched mRNA. In contrast, four of 11 sporadic basal cell carcinomas, that were examined for comparison, showed loss of heterozygosity at the patched gene locus (p <0.05), and moderate to strong signals for patched mRNA was observed in all seven basal cell carcinoma tumors examined (p <0.0001). Additional investigation by reverse transcription polymerase chain reaction in four basal cell carcinomas and two nevus sebaceous tumors also showed the expression of Gli-1, another target gene in the patched hedgehog signaling pathway, in all the basal cell carcinomas samples but not in any of the nevus sebaceous tumors examined. The findings in this study do not support the view that the deregulation of the patched-hedgehog signaling pathway is involved in the pathogenesis of nevus sebaceous and associated tumors, and show that, although morphologically similar, trichoblastomas and basal cell carcinomas have a different molecular pathogenesis. PMID- 11886540 TI - Lack of association between NOD2 3020InsC frameshift mutation and psoriasis. PMID- 11886541 TI - XP43TO, previously classified as xeroderma pigmentosum Group E, should be reclassified as xeroderma pigmentosum variant. PMID- 11886542 TI - A simple in vivo system for studying epithelialization, hair follicle formation, and invasion using primary epidermal cells from wild-type and transgenic ornithine decarboxylase-overexpressing mouse skin. PMID- 11886543 TI - Late Breaking Abstracts for the 31st European Society for Dermatological Research (ESDR) Meeting 2001 Stockholm, Sweden September, 20-22, 2001. PMID- 11886548 TI - A three-protein inhibitor of polar septation during sporulation in Bacillus subtilis. AB - We present evidence for a three-protein inhibitor of polar division that locks in asymmetry after the formation of a polar septum during sporulation in Bacillus subtilis. Asymmetric division involves the formation of cytokinetic Z-rings near both poles of the developing cell. Next, a septum is formed at one of the two polar Z-rings, thereby generating a small, forespore cell and a mother cell. Gene expression under the control of the mother-cell transcription factor sigmaE is needed to block cytokinesis at the pole distal to the newly formed septum. We report that this block in polar cytokinesis is mediated partly by sigmaE-directed transcription of spoIID, spoIIM and spoIIP, sporulation genes that were known to be involved in the subsequent process of forespore engulfment. We find that a spoIID, spoIIM and spoIIP triple mutant substantially mimicked the bipolar division phenotype of a sigmaE mutant and that cells engineered to produce SpoIID, SpoIIM and SpoIIP prematurely were inhibited in septum formation at both poles. Consistent with the hypothesis that SpoIID, SpoIIM and SpoIIP function at both poles of the sporangium, a GFP--SpoIIM fusion localized to the membrane that surrounds the engulfed forespore and to the potential division site at the distal pole. PMID- 11886549 TI - Systematic mutational analysis of the amino-terminal domain of the Listeria monocytogenes ActA protein reveals novel functions in actin-based motility. AB - The Listeria monocytogenes ActA protein acts as a scaffold to assemble and activate host cell actin cytoskeletal factors at the bacterial surface, resulting in directional actin polymerization and propulsion of the bacterium through the cytoplasm. We have constructed 20 clustered charged-to-alanine mutations in the NH2-terminal domain of ActA and replaced the endogenous actA gene with these molecular variants. These 20 clones were evaluated in several biological assays for phenotypes associated with particular amino acid changes. Additionally, each protein variant was purified and tested for stimulation of the Arp2/3 complex, and a subset was tested for actin monomer binding. These specific mutations refined the two regions involved in Arp2/3 activation and suggest that the actin binding sequence of ActA spans 40 amino acids. We also identified a 'motility rate and cloud-to-tail transition' region in which nine contiguous mutations spanning amino acids 165-260 caused motility rate defects and changed the ratio of intracellular bacteria associated with actin clouds and comet tails without affecting Arp2/3 activation. Several unusual motility phenotypes were associated with amino acid changes in this region, including altered paths through the cytoplasm, discontinuous actin tails in host cells and the tendency to 'skid' or dramatically change direction while moving. These unusual phenotypes illustrate the complexity of ActA functions that control the actin-based motility of L. monocytogenes. PMID- 11886550 TI - Distribution of the Escherichia coli structural maintenance of chromosomes (SMC) like protein MukB in the cell. AB - Fluorescent polyclonal antibodies specific for MukB have been used to study its localization in Escherichia coli. In wild-type cells, the MukB protein appeared as a limited number of oblong shapes embracing the nucleoid. MukB remained associated with the nucleoid in the absence of DNA replication. The centre of gravity of the dispersed MukB signal initially localized near mid-cell, but moved to approximately quarter positions well before the termination of DNA replication and its subsequent reinitiation. Because MukB had been reported to bind to FtsZ and to its eukaryotic homologue tubulin in vitro, cells were co-labelled with MukB- and FtsZ-specific fluorophores. No co-localization of MukB with polymerized FtsZ (the FtsZ ring) was observed at any time during the cell cycle. A possible role for MukB in preventing premature FtsZ polymerization and in DNA folding that might assist DNA segregation is discussed. PMID- 11886551 TI - The incompatibility between the PlcR- and AtxA-controlled regulons may have selected a nonsense mutation in Bacillus anthracis. AB - Bacillus anthracis, Bacillus thuringiensis and Bacillus cereus are members of the Bacillus cereus group. These bacteria express virulence in diverse ways in mammals and insects. The pathogenic properties of B. cereus and B. thuringiensis in mammals results largely from the secretion of non-specific toxins, including haemolysins, the production of which depends upon a pleiotropic activator PlcR. In B. anthracis, PlcR is inactive because of a nonsense mutation in the plcR gene. This suggests that the phenotypic differences between B. anthracis on the one hand and B. thuringiensis and B. cereus on the other could result at least partly from loss of the PlcR regulon. We expressed a functional PlcR in B. anthracis. This resulted in the transcriptional activation of genes weakly expressed in the absence of PlcR. The transcriptional activation correlated with the induction of enzymatic activities and toxins including haemolysins. The toxicity of a B. anthracis PlcR+ strain was assayed in the mouse subcutaneous and nasal models of infection. It was no greater than that of the parental strain, suggesting that the PlcR regulon has no influence on B. anthracis virulence. The PlcR regulon had dramatic effects on the sporulation of a B. anthracis strain containing the virulence plasmid pXO1. This resulted from incompatible interactions with the major AtxA-controlled virulence regulon. We propose that the PlcR-controlled regulon in B. anthracis has been counterselected on account of its disadvantageous effects. PMID- 11886552 TI - The sporulation transcription factor Spo0A is required for biofilm development in Bacillus subtilis. AB - Biofilms are structured communities of cells encased in a polymeric matrix and adherent to a surface, interface or each other. We report here that the soil bacterium Bacillus subtilis forms biofilms. By confocal scanning laser microscopy, we observed that B. subtilis adhered to abiotic surfaces and formed a three-dimensional structure > or =30 microm in depth. These biofilms appeared to be at least partly encased in an extracellular polysaccharide matrix, as they could be stained with Calcofluor, a polysaccharide-binding dye. To understand the molecular mechanism of biofilm formation, we screened previously characterized mutants for a defect in biofilm formation. We found that mutations in spo0A, which encodes the major early sporulation transcription factor, caused a defect in biofilm formation. spo0A mutant cells adhered to a surface in a monolayer of cells rather than a three-dimensional biofilm. The requirement of Spo0A for biofilm development appears to result from its role in negatively regulating AbrB. Mutations in abrB suppressed the biofilm defect of a spo0A mutant, indicating that AbrB negatively regulates at least one gene that is required for the transition from a monolayer of attached cells to a mature biofilm. Implications of biofilm development for the ecology of B. subtilis are discussed. PMID- 11886553 TI - Isolation and characterization of topological specificity mutants of minD in Bacillus subtilis. AB - In rod-shaped bacteria such as Bacillus subtilis, division site selection is mediated by MinC and MinD, which together function as a division inhibitor. Topological specificity is imposed by DivIVA, which ensures that MinCD specifically inhibits division close to the cell poles, while allowing division at mid-cell. MinD plays a central role in this process, as it positions and activates MinC and is dependent on DivIVA for its own positioning at the poles. To investigate MinD activities further, we have constructed and analysed a collection of minD mutants. Mutations in the conserved ATPase motifs lead to an inactive protein, possibly unable to oligomerize, but which nevertheless retains some affinity for the cell membrane. Several mutations affecting the mid- to C terminal parts of MinD led to a protein probably unable to interact with DivIVA, but that could still stimulate division inhibition by MinC. These findings suggest that the ATPase activity of MinD is necessary for all its functions (possibly in part by controlling the oligomerization state of the protein). The other mutations may identify a surface of MinD involved in its interactions with DivIVA and a possible mechanism for control of MinD by DivIVA. PMID- 11886554 TI - Spontaneous sequence duplication within an open reading frame of the pneumococcal type 3 capsule locus causes high-frequency phase variation. AB - The molecular genetic basis of high-frequency serotype 3 capsule phase variation in Streptococcus pneumoniae (the pneumococcus) was investigated. Pneumococci were grown in sorbarod biofilms at 34 degrees C to mimic nasopharyngeal carriage. Different type 3 pneumococci commonly associated with invasive disease generated apparently random tandem duplications of 11-239 bp segments within the cap3A gene of the type 3 capsule locus. These duplications alone were found to be responsible for high-frequency capsule phase variation, in which (phase off) acapsular variants possessed duplications within cap3A, and (phase on) capsular revertants possessed wild-type cap3A genes, indicating the precise excision of the duplication. Additionally, the frequency of phase reversion (off to on) was found to exhibit a linear relationship between (log) frequency of reversion and (log) length of duplication. This apparently random duplication giving rise to phase variation is in stark contrast to the 'preprogrammed' contingency genes in many Gram-negative organisms that possess homopolymeric sequence repeats or motifs for site-specific recombination. PMID- 11886555 TI - Sister chromosome cohesion of Escherichia coli. AB - We analysed Escherichia coli cells synchronized for initiation of chromosomal DNA replication by fluorescence in situ hybridization (FISH) using fluorescent DNA probes corresponding to various chromosomal regions. Sister copies of regions in an approximately oriC-proximal half of the chromosome are cohesive with each other after replication until the late period of chromosome replication. Sister copies of regions relatively close to the terminus are also separated from each other in the same late period of replication. It is important that sister copies in all the tested regions are thus separated from each other nearly all at once in the late period of chromosome replication. These results are consistent with results obtained by FISH in randomly growing cultures. Cohesion of sister copies in an oriC-close region is observed in a dam null mutant lacking DNA adenine methyltransferase the same as in the parental isogenic dam+ strain, indicating that the cohesion is independent of DNA adenine methyltransferase. This further implies that hemimethylated DNA-binding proteins, such as SeqA, are not involved in the cohesion. On the other hand, the cohesion of sister copies of the oriC close region was not observed in mukB null mutant cells, suggesting that MukB might be involved in the chromosome cohesion. PMID- 11886556 TI - Distinct and redundant roles of the two protein kinase A isoforms Tpk1p and Tpk2p in morphogenesis and growth of Candida albicans. AB - TPK1 and TPK2 encode both isoforms of protein kinase A (PKA) catalytic subunits in Candida albicans. Mutants lacking both TPK1 alleles showed defective hyphal morphogenesis on solid inducing media, whereas in liquid hypha, formation was affected slightly. In contrast, tpk2 mutants were only partially morphogenesis defective on solid media, whereas a strong block was observed in liquid. In addition, the yeast forms of tpk2-- but not tpk1-- mutants were completely deficient in invading agar. Because Tpk1p and Tpk2p differ in their N-terminal domains of approximately 80--90 amino acids, while the catalytic portions are highly homologous, the functions of hybrid Tpk proteins with exchanged N-terminal domains were tested. The results demonstrate that the catalytic portions mediate Tpk protein specificities with regard to filamentation, whereas agar invasion is mediated by the N-terminal domain of Tpk2p. Homozygous tpk1 and tpk2 mutants grew normally; however, a tpk2 mutant strain containing a single regulatable TPK1 allele (PCK1p-TPK1) at low expression levels was severely growth defective. It was completely blocked in hyphal morphogenesis and was stress resistant to high osmolarities or temperatures. Thus, both Tpk isoforms in C. albicans share growth functions but, unlike Saccharomyces cerevisiae isoforms, they have positive, specific roles in filament formation in different environments. PMID- 11886557 TI - Overexpression of the alternative oxidase restores senescence and fertility in a long-lived respiration-deficient mutant of Podospora anserina. AB - Several lines of evidence have implicated reactive oxygen species (ROS) in the pathogenesis of various degenerative diseases and in organismal ageing. Furthermore, it has been shown recently that the alternative pathway respiration present in plants lowers ROS mitochondrial production. An alternative oxidase (AOXp) also occurs in the filamentous fungus Podospora anserina. We show here that overexpression of this oxidase does not decrease ROS production and has no effect on longevity, mitochondrial stability or ageing in this fungus. In the same way, inactivation of the gene has no effect on these parameters. In contrast, overexpression of the alternative oxidase in the long-lived cox5::BLE mutant, deficient in cytochrome c oxidase, considerably increases ROS production of the mutant. It rescues slow growth rate and female sterility, indicating an improved energy level. This overexpression also restores senescence and mitochondrial DNA instability, demonstrating that these parameters are controlled by the energy level and not by the expression level of the alternative oxidase. We also suggest that expression of this oxidase in organisms naturally devoid of it could rescue respiratory defects resulting from cytochrome pathway dysfunctions. PMID- 11886558 TI - Phosphoserine modification of the enteropathogenic Escherichia coli Tir molecule is required to trigger conformational changes in Tir and efficient pedestal elongation. AB - Enteropathogenic Escherichia coli (EPEC) virulence is correlated with intimate adherence to gut epithelial cells, loss of absorptive microvilli and reorganization of host cytoskeletal proteins into pedestal-like structures beneath the adherent bacteria. These processes depend on Tir (i) being inserted into the plasma membrane; (ii) being tyrosine phosphorylated; and (iii) interacting with the bacterial outer membrane protein, intimin. However, phosphorylation on other undefined residues leads to approximately 5 kDa and approximately 2 kDa increases in Tir apparent molecular mass within host cells. In this study, we show that equivalent shifts can be induced in vitro by phosphorylation of Tir on two serine (S434 and S463) residues by protein kinase A (PKA). Our data suggest that the sequential addition of two phosphate groups triggers conformational changes in Tir structure that may supply the energy to insert Tir into the plasma membrane. PKA was also shown to modify Tir within host cells on S434 to induce the approximately 5 kDa shift. Whereas modification of S434 was not essential to generate an actin-nucleating molecule, it was required for Tir to induce pedestal elongation efficiently. This study not only increases our understanding of the mechanism by which phosphorylation induces shifts in Tir apparent molecular mass and suggests a mechanism by which Tir may be inserted into the plasma membrane, but also reveals a role for non-tyrosine phosphorylation in Tir function and identifies the first kinase that can modify Tir in vitro or in vivo. PMID- 11886559 TI - Sensing nitrogen limitation in Corynebacterium glutamicum: the role of glnK and glnD. AB - A novel nitrogen control system regulating the transcription of genes expressed in response to nitrogen starvation in Corynebacterium glutamicum was identified by us recently. In this communication, we also show that the nitrogen regulation cascade in C. glutamicum functions by a new mechanism, although components highly similar to sensor and signal transmitter proteins of Escherichia coli are used, namely uridylyltransferase and a PII-type GlnK protein. The genes encoding these key components of the nitrogen regulation cascade, glnD and glnK, are organized in an operon together with amtB, which codes for an ammonium permease. Using a combination of site-directed mutagenesis, RNA hybridization experiments, reporter gene assays, transport measurements and non-denaturing gel electrophoresis followed by immunodetection, we showed that GlnK is essential for nitrogen control and that signal transduction is transmitted by uridylylation of this protein. As a consequence of the latter, a glnD deletion strain lacking uridylyltransferase is impaired in its response to nitrogen shortage. The glnD mutant revealed a decreased growth rate in the presence of limiting amounts of ammonium or urea; additionally, changes in its protein profile were observed, as shown by in vivo labelling and two-dimensional PAGE. In contrast to E. coli, expression of glnD is upregulated upon nitrogen limitation in C. glutamicum. This indicates that the glnD gene product is probably not the primary sensor of nitrogen status in C. glutamicum as shown for enterobacteria. In accordance with this hypothesis, we found a deregulated nitrogen control as a result of the overexpression of glnD. Furthermore, quantification of cytoplasmic amino acid pools excluded the possibility that a fall in glutamine concentration is perceived as the signal for nitrogen starvation by C. glutamicum, as is found in enterobacteria. Direct measurements of the intracellular ammonium pool indicated that the concentration of this compound might indicate the cellular nitrogen status. Deduced from glnK and glnD expression patterns and the genetic organization of these genes, this regulatory mechanism is also present in Corynebacterium diphtheriae, the causative agent of diphtheria. PMID- 11886560 TI - The Fur repressor controls transcription of iron-activated and -repressed genes in Helicobacter pylori. AB - The ferric uptake regulator (Fur) protein is known to act as a Fe2+-dependent transcriptional repressor of bacterial promoters. Here, we show that, in Helicobacter pylori, Fur can mediate the regulation of iron-activated genes in contrast to classical Fur regulation, in which iron acts as a co-repressor. Inactivation of the fur gene in the chromosome of H. pylori resulted in the derepression of a 19 kDa protein that was identified by N-terminal sequencing as the non-haem-containing ferritin (Pfr). Growth of the wild-type H. pylori strain on media treated with increasing concentrations of FeSO4 resulted in induction of transcription from the Ppfr promoter and, conversely, depletion of iron resulted in repression of Ppfr, indicating that this promoter is iron activated. In the fur mutant, the Ppfr promoter is constitutively highly expressed and no longer responds to iron, indicating that the Fur protein mediates this type of iron regulation. Footprinting analysis revealed that Fur binds to the Ppfr promoter region and that Fe2+ decreases the efficiency of binding. In contrast, Fe2+ increased the affinity of Fur for a classical Fur-regulated promoter, the iron repressed frpB gene promoter. To our knowledge, this is the first evidence of direct interaction between the Fur protein and the promoter of an iron-activated (-derepressed) gene. Our results support a model in which the iron status of the Fur protein differentially alters its affinity for operators in either iron repressed or iron-activated genes. PMID- 11886561 TI - A molecular mechanism for the repression of transcription by the H-NS protein. AB - The H-NS protein is a major component of the bacterial nucleoid and plays a crucial role in the global gene regulation of enteric bacteria. Although H-NS does not exhibit a high DNA sequence specificity, a number of H-NS-responsive promoters have been shown to contain regions of intrinsic DNA curvature located either upstream or downstream of the transcription start point. We have studied H NS binding to DNA and in vitro transcriptional regulation by H-NS at several synthetic promoters with or without curved sequences inserted upstream of the Pribnow box. We show how such inserts determine the final organization of H-NS containing nucleoprotein complexes and how this affects transcription. We refine a two-step mechanism for the constitution of H-NS assemblies that are efficient in regulation. PMID- 11886562 TI - In vivo aggregation of the HET-s prion protein of the fungus Podospora anserina. AB - We have proposed that the [Het-s] infectious cytoplasmic element of the filamentous fungus Podospora anserina is the prion form of the HET-s protein. The HET-s protein is involved in a cellular recognition phenomenon characteristic of filamentous fungi and known as heterokaryon incompatibility. Under the prion form, the HET-s protein causes a cell death reaction when co-expressed with the HET-S protein, from which it differs by only 13 amino acid residues. We show here that the HET-s protein can exist as two alternative states, a soluble and an aggregated form in vivo. As shown for the yeast prions, transition to the infectious prion form leads to aggregation of a HET-s--green fluorescent protein (GFP) fusion protein. The HET-s protein is aggregated in vivo when highly expressed. However, we could not demonstrate HET-s aggregation at wild-type expression levels, which could indicate that only a small fraction of the HET-s protein is in its aggregated form in vivo in wild-type [Het-s] strains. The antagonistic HET-S form is soluble even at high expression level. A double amino acid substitution in HET-s (D23A P33H), which abolishes prion infectivity, suppresses in vivo aggregation of the GFP fusion. Together, these results further support the model that the [Het-s] element corresponds to an abnormal self perpetuating aggregated form of the HET-s protein. PMID- 11886563 TI - Systematic mutagenesis of the Helicobacter pylori cag pathogenicity island: essential genes for CagA translocation in host cells and induction of interleukin 8. AB - Helicobacter pylori (Hp) carries a type IV secretion system encoded by the cag pathogenicity island (cag-PAI), which is used to: (i) translocate the bacterial effector protein CagA into different types of eukaryotic cells; and (ii) induce the synthesis and secretion of chemokines, such as interleukin-8 (IL-8). The cag PAI in Hp 26695 consists of 27 putative genes, six of which were identified as homologues to the basic type IV secretion system represented by the Agrobacterium tumefaciens virB operon. To define the role and contribution of each of the 27 genes, we applied a precise deletion/insertion mutagenesis procedure to knock out each individual gene without causing polar effects on the expression of downstream genes. Seventeen out of 27 genes were found to be absolutely essential for translocation of CagA into host cells and 14 out of 27 for the ability of Hp fully to induce transcription of IL-8. The products of hp0524 (virD4 homologue), hp0526 and hp0540 are absolutely essential for the translocation of CagA, but not for the induction of IL-8. In contrast, the products of hp0520, hp0521, hp0534, hp0535, hp0536 and hp0543 are not necessary for either translocation of CagA or for IL-8 induction. Our data argue against a translocated IL-8-inducing effector protein encoded by the cag-PAI. We isolated a variant of Hp 26695, which spontaneously switched off its capacity for IL-8 induction and translocation of CagA, but retained the complete cag-PAI. We identified a point mutation in gene hp0532, causing a premature translational stop in the corresponding polypeptide chain, providing a putative explanation for the defect in the type IV secretion system of the spontaneous mutant. PMID- 11886564 TI - Interactions of the Trichoderma reesei rho3 with the secretory pathway in yeast and T. reesei. AB - We recently isolated from the filamentous fungus Trichoderma reesei (Hypocrea jecorina) a gene encoding RHOIII as a multicopy suppressor of the yeast temperature-sensitive secretory mutation, sec15-1. To characterize this gene further, we tested its ability to suppress other late-acting secretory mutations. The growth defect of yeast strains with sec1-1, sec1-11, sec3-2, sec6-4 and sec8 9 mutations was suppressed. Expression of rho3 also improved the impaired actin organization of sec15-1 cells at +38 degrees C. Overproduction of yeast Rho3p using the same expression vector as T. reesei RHOIII appeared to be toxic in sec3 101, sec5-24, sec8-9, sec10-2 and sec15-1 cells. When expressed from the GAL1 promoter, RHO3 suppressed the growth defect of sec1 at the restrictive temperature and inhibited the growth of sec3-101 at the permissive temperature. Disruption of the rho3 gene in the T. reesei genome did not affect the hyphal or colony morphology nor the cellular cytoskeleton organization. Furthermore, the growth of T. reesei was not affected on glucose by the rho3 disruption. Instead, both growth and protein secretion of T. reesei in cellulose cultures was remarkably decreased in rho3 disruptant strains when compared with the parental strain. These results suggest that rho3 is involved in secretion processes in T. reesei. PMID- 11886565 TI - Pairing of P1 plasmid partition sites by ParB. AB - The mechanisms by which bacterial plasmids and chromosomes are partitioned are largely obscure, but it has long been assumed that the molecules to be separated are initially paired, as are sister chromatids in mitosis. We offer in vivo evidence that the partition protein ParB encoded by the bacterial plasmid P1 can pair cis-acting partition sites of P1 inserted in a small, multicopy plasmid. ParB was shown previously to be capable of extensive spreading along DNA flanking the partition sites. Experiments in which ParB spreading was constrained by physical roadblocks suggest that extensive spreading is not required for the pairing process. PMID- 11886566 TI - Replication arrests during a single round of replication of the Escherichia coli chromosome in the absence of DnaC activity. AB - We used a flow cytometric assay to determine the frequency of replication fork arrests during a round of chromosome replication in Escherichia coli. After synchronized initiation from oriC in a dnaC(Ts) strain, non-permissive conditions were imposed, such that active DnaC was not available during elongation. Under these conditions, about 18% of the cells failed to complete chromosome replication. The sites of replication arrests were random and occurred on either arm of the bidirectionally replicating chromosome, as stalled forks accumulated at the terminus from both directions. The forks at the terminal Ter sites disappeared in the absence of Tus protein, as the active forks could then pass through the terminus to reach the arrest site, and the unfinished rounds of replication would be completed without DnaC. In a dnaC2(Ts)rep double mutant, almost all cells failed to complete chromosome replication in the absence of DnaC activity. As inactivation of Rep helicase (the rep gene product) has been shown to cause frequent replication arrests inducing double-strand breaks (DSBs) in a replicating chromosome, DnaC activity appears to be essential for replication restart from DSBs during elongation. PMID- 11886569 TI - Therapeutic apheresis: current perspectives. PMID- 11886570 TI - Myocardial infarction/injury is relatively common at presentation of acute thrombotic thrombocytopenic purpura: the Indiana University experience. AB - Although widespread vascular thrombosis is common in thrombotic thrombocytopenic purpura (TTP), there have been no prospective studies on the extent of injury to specific organs. Following successful resuscitation and plasma exchange of an index patient with widespread organ dysfunction, cardiogenic shock, and elevated cardiac troponin-I levels, we prospectively studied and identified 2 more individuals (of 10 consecutive patients) with evidence of myocardial injury/infarction at presentation of acute TTP. These data suggest that cardiac troponin-I measurements should be considered during initial evaluation of all patients with acute TTP. PMID- 11886571 TI - Peripheral blood progenitor cell transplantation. AB - Peripheral blood progenitor cells (PBPCs) have become increasingly popular over the last 15 years as the source of hematopoietic stem cells for transplantation. In the early 1990s, PBPCs replaced bone marrow (BM) as the preferred source of autologous stem cells, and recently the same phenomenon is seen in the allogeneic setting. Under steady-state conditions, the concentration of PBPCs (as defined by CFU-GM and/or CD34+ cells) is very low, and techniques were developed to increase markedly this concentration. Such mobilization techniques include daily injections of filgrastim (G-CSF) or a combination of chemotherapy and growth factors. Leukapheresis procedures allow the collection of large numbers of circulating white blood cells (and PBPCs). One or two leukapheresis procedures are often sufficient to obtain the minimum number of CD34+ cells considered necessary for prompt and consistent engraftment (i.e., 2.5-5.0 x 10(6)/kg). As compared to BM, autologous transplants with PBPCs lead to faster hematologic recovery and have few, if any, disadvantages. In the allogeneic arena, PBPCs also result in faster engraftment, but at a somewhat higher cost of chronic graft versus-host disease (GvHD). This may be a double-edged sword leading to both increased graft-versus-tumor effects and increased morbidity. The rapid advances in the study of hematopoietic, and even earlier, stem cells will continue to shape the future of PBPC transplantation. PMID- 11886572 TI - Leukocytoreduction for acute leukemia. AB - Both in children and adults, acute leukemia may present with extremely high blast counts; a phenomenon known as hyperleukocytosis. Respiratory failure, intracranial bleeding, and severe metabolic abnormalities frequently occur in acute hyperleukocytic leukemias (AHLs) and are the primary determinants of the high early mortality (20% to 40%) observed. The process leading to these complications has long been known as leukostasis, but the biological mechanisms underlying its development and progression have remained unclear. Traditionally, leukostasis has been attributed to overcrowding of leukemic blasts in the microcirculation, and its treatment has focused on prompt leukocytoreduction. However, it is becoming increasingly evident that leukostasis results from the adhesive interactions between leukemic blasts and the endothelium; a mechanism that none of the current therapies directly addresses. The endothelial damage associated with leukostasis is likely to be mediated by cytokines released in situ and by subsequent migration of leukemic blasts in the perivascular space. The adhesion molecules displayed by the leukemic blasts and their chemotactic response to the cytokines in the vascular microenvironment are probably more important in causing leukostasis than the cell number. This may explain why leukostasis may develop in some patients with AHL and not in others, and why some patients with acute leukemia without hyperleukocytosis (<50,000 blasts/mm(3)) develop leukostasis and respond to leukocytoreduction. Leukapheresis effectively reduces the blast count in many patients with AHL and is routinely used for immediate leukocytoreduction. However, the most appropriate use of leukapheresis in acute leukemia remains unclear, and the procedure may not prevent early death more efficiently than fluid therapy, hydroxyurea, and prompt induction chemotherapy. The use of cranial irradiation remains very controversial and is not generally recommended. The identification of the adhesion molecules, soluble cytokines, and chemotactic ligand-receptor pairs mediating endothelial cell damage in AHL should become a priority if better outcomes are desired. PMID- 11886573 TI - The role of red blood cell exchange transfusion in the treatment and prevention of complications of sickle cell disease. AB - Patients with sickle cell disease have abnormal red blood cells (RBCs). This can cause chronic hemolytic anemia and vaso-occlusion leading to tissue hypoxemia and organ dysfunction. RBC exchange transfusion can, without increasing the whole blood viscosity, quickly replace abnormal erythrocytes with normal and raise the hematocrit resulting in improved delivery of oxygen to hypoxic tissues. Unfortunately, transfusion can also be associated with complications. This paper reviews the role of transfusion, both simple and exchange, in the treatment and prevention of sickle-related complications. The benefits of exchange versus simple transfusion and transfusion versus alternative therapies are discussed. PMID- 11886574 TI - Neonatal alloimmune thrombocytopenia. AB - Neonatal alloimmune thrombocytopenia (NAIT) can occur when a mother is immunized against fetal platelet antigens inherited from the father. Early diagnosis and appropriate platelet transfusion therapy are essential to prevent life threatening intracranial hemorrhage in the thrombocytopenic fetus or neonate. Five major human platelet antigen (HPA) systems are capable of causing this disorder with HPA-1a indicated most frequently. This article reviews the pathophysiology, clinical aspects, and management of NAIT. We also present our experience with treatment of neonates affected with this disorder. PMID- 11886575 TI - The role of blood component removal in essential and reactive thrombocytosis. AB - An elevated platelet count is now a common finding in both hospitalized and ambulatory patients with the advent of automated complete blood cell counters. Clinicians may be called upon to make a distinction between a reactive process and a primary hematologic disorder as the cause of a thrombocytosis and to determine whether treatment is indicated. Essential thrombocythemia and other myeloproliferative disorders may present with marked increases in the platelet counts and may be associated with thrombohemorrhagic complications. Reactive thrombocytosis can be caused by iron deficiency and a variety of inflammatory conditions, infections, malignancy, bleeding or hemolysis, splenectomy, and drugs. Acute therapy for all of these disorders has included blood component removal, specifically plateletpheresis. The role of plateletpheresis in current management of thrombocytosis is considered, based on current knowledge of pathophysiology and a review of the literature. PMID- 11886576 TI - Plasmapheresis in the dysproteinemias. AB - The dysproteinemias consist of a broad range of serious disease states with the common thread of excessive production of an abnormal, or para-protein. Various clinical syndromes may arise, either from the underlying disease process, the excess paraprotein, or both. Clinical presentation depends upon the organ system(s) affected by the abnormal protein. Diseases included under the classification of dysproteinemias include cryoprotein-related diseases, Waldenstrom's macroglobulinemia, hyperviscosity syndrome, monoclonal gammopathy, multiple myeloma, light chain disease, and amyloidosis. Plasmapheresis, often in conjunction with other therapies, has been widely used to treat the dysproteinemias and their resulting clinical syndromes. Automated plasmapheresis, which separates plasma from the cellular blood elements by centrifugation, is used most commonly in the United States. Membrane separation and immunoadsorption techniques are more commonly used in Europe and Japan. In automated plasmapheresis, the plasma is removed from the patient's circulation and replaced with a protein-based fluid such as 5% human albumin solution or plasma protein fraction or with fresh frozen plasma. Membrane separation and immunoadsorption allow the offending proteins to be removed more selectively from the patient's plasma prior to the plasma being returned to the patient. This review article presents a description of each disease, the rationale for plasmapheresis therapy, recommended schedules of plasmapheresis, and the use of adjunctive therapies. Results of published studies, case reports, and the author's experience in treating these diseases will serve as the foundation for discussion. PMID- 11886577 TI - Apheresis in treatment of the inflammatory demyelinating peripheral neuropathies. AB - The inflammatory demyelinating peripheral neuropathies are recognizable treatable entities within the broad field of diseases of the peripheral nervous system. Most peripheral nerve diseases are still of undetermined causation and have no direct disease-modifying treatment. This paper reviews the diagnostic features of the inflammatory neuropathies, presumed mechanisms of pathogenesis, and the role of apheresis, along with other immunomodulating therapies, in the management of these diseases. PMID- 11886578 TI - Myasthenia gravis and Lambert-Eaton syndrome. AB - Myasthenia gravis is a common autoimmune disorder characterized by the presence of pathogenic antibodies directed against the acetylcholine receptor. Patients present with variable degrees and distribution of fluctuating weakness at times life threatening. Clinical manifestations, establishment of diagnosis, the natural history of myasthenia gravis, and therapeutic options are herein reviewed. Far less common is Lambert-Eaton syndrome (the myasthenic syndrome), another autoimmune disorder due to the presence of antibodies directed against the PQ-type voltage-gated calcium channels. Clinical features and treatment options are summarized. PMID- 11886579 TI - Apheresis in cryoglobulinemia complicating hepatitis C and in other renal diseases. AB - Removal of cryoglobulins by plasma exchange is now an accepted therapy. Cryoglobulins are circulating complexes that can deposit on small vessels and cause limited or extensive tissue injury. There are 3 major classes of cryoglobulins. Type I cryoglobulins are monoclonal and are detected in a variety of lymphoproliferative disorders. Type II cryoglobulins are mixed containing monoclonal and polyclonal IgG or IgM molecules. Type III cryoglobulins are also mixed and contain polyclonal IgG. Type II cryoglobulins are largely caused by hepatitis C virus infection; hence, they are the most common of the 3 types. In hepatitis C, cryoglobulins are linked to glomerular immune complex injury, often times accompanied by vasculitis of the skin, nerves, and other vital organs. Immediate removal of cryoglobulins by plasma exchange is an effective short-term treatment that can complement more-specific therapies. Plasma exchange has also been used to remove other circulating nephrotoxic agents such as antiglomerular basement antibodies that cause Goodpasture's syndrome, protease inhibitor autoantibodies that cause thrombotic thrombocytopenic purpura, and antiglomerular factors that cause some types of focal glomerulosclerosis. PMID- 11886580 TI - Volunteer donor apheresis. AB - Volunteer donor apheresis has evolved from early plasmapheresis procedures that collected single components into technically advanced multicomponent procedures that can produce combinations of red blood cells, platelets, and plasma units. Blood collection and utilization is increasing annually in the United States. The number of apheresis procedures is also increasing such that single donor platelet transfusions now exceed platelet concentrates from random donors. Donor qualifications for apheresis vary from those of whole blood. Depending on the procedure, the donor weight, donation interval, and platelet count must be taken into consideration. Adverse effects of apheresis are well known and fortunately occur in only a very small percentage of donors. The recruitment of volunteer donors is one of the most challenging aspects of a successful apheresis program. As multicomponent apheresis becomes more commonplace, it is important for collection centers to analyze the best methods to recruit and collect donors. PMID- 11886581 TI - Influence of storage time on activation of platelets collected with CS 3000 Plus and Cobe Spectra using platelet storage containers. AB - The aim of this study was to evaluate the in vitro activation in platelet suspensions collected with CS 3000 Plus and Cobe Spectra cell separators using platelet storage containers and the role of white blood cell (WBC) concentration of the suspension in this activation. Seventy-seven donors were subjected to automated platelet donations with 1 type of equipment (37 with Cobe Spectra and 40 with CS 3000 Plus). Blood samples were obtained immediately after separation and on the third day of storage at 22 degrees C in constant agitation. The WBC concentrations of these samples were studied before storage. Paraformaldehyde fixed platelets were incubated with 2 murine monoclonal antibodies: CD42b and CD62. Murine monoclonal antibody immunoglobulin G was used as a negative isotypic control. Bound antibody was then quantitated by flow cytometry. On the third day of storage, a significant increase in CD62 expression rate was observed in platelet suspensions collected with both kinds of equipment. Mean expression rates for Cobe Spectra on Day 0 and Day 3 were 25.6 +/- 6.2% and 69.2 +/- 9.7%, respectively. Mean expression rates for CS 3000 Plus on Day 0 and Day 3 were 23.4 +/- 8.2% and 67.0 +/- 8.2%, respectively. The mean results for both devices were 22.8 +/- 4.56% for Day 0 and 68.7 +/- 13.2% for Day 3. There was no difference between CD42b mean fluorescence intensity on Days 0 and 3 for the 2 devices (p > 0.5). Mean WBC concentrations in the platelet suspensions for Cobe Spectra and CS 3000 Plus were 0.37 x 10(3)/microl and 0.42 x 10(3)/microl, respectively, and there was no relation between WBC concentration and increase in CD62 expression. Both kinds of equipment were found to be similar according to in vitro activation markers. PMID- 11886582 TI - Performance evaluation of plasma fractionation membrane. AB - Membrane plasma fractionation is an important part of double filtration plasmapheresis. We evaluated a newly developed polyethersulfone (PES) hollow fiber plasma fractionation membrane, using 3 parameters: abnormal protein ratio (APRR), different protein ratio (DPR), and plasma filtration efficiency, except sieving coefficients. The APRR of the fractionation membrane was over 0.90, and the DPR changed after the plasma fractionation. We also compared the sieving coefficients of the PES hollow membrane with some commercial plasma fractionation membranes. The data were significant for the clinical use of the PES hollow-fiber fractionation membrane. PMID- 11886583 TI - Continuous arterial infusion of prostaglandin E(1) via the superior mesenteric artery in the treatment of postoperative liver failure. AB - Impaired hepatic blood flow is one of the causative factors in postoperative liver failure. To restore the hepatic blood flow in case of hepatic artery interruption (HAI), the effect of continuous arterial infusion of prostaglandin E(1) (PGE(1)), which has a strong vasodilatory effect on vascular smooth muscles, was assessed experimentally and clinically. Twelve pigs underwent ligation and division of the hepatic artery and were divided into 2 groups. In the control group, saline was infused in the superior mesenteric artery (SMA), and in the PGE(1) group, 0.02 microg/kg/min of PGE(1) was infused continuously in the SMA. Hepatic oxygen delivery (HDO(2)) in the control group was 87.8 +/- 8.9 ml/min before HAI and decreased to 43.1 +/- 2.6 ml/min at 60 min after HAI, showing 50.9% decrease by HAI. On the contrary, HDO(2) in the PGE(1) group was 86.7 +/- 9.1 ml/min before HAI and was 76.6 +/- 12.2 ml/min at 60 min after HAI, showing only 11.6% decrease by HAI. Clinically, a 65-year-old female suffering from cholangiocellular carcinoma underwent extended left hepatic lobectomy. At operation, the branch of the hepatic artery to the anterior segment of the liver was ligated, and the right branch of the portal vein became stenotic unavoidably. Postoperatively, severe liver dysfunction developed so that continuous PGE1 infusion in the SMA was initiated at a rate of 0.01 microg/kg/min on the eighth postoperative day and continued for 9 days. Plasma exchange was performed twice concomitantly. Portal venous flow increased from 612 ml/min to 1,192 ml/min, and bile flow from external biliary drainage tube doubled by the PGE(1) infusion. The liver function was ameliorated after PGE(1) infusion. PMID- 11886584 TI - A case report: first case of filtration leukocytapheresis for a patient of aortitis syndrome associated with ulcerative colitis. AB - An 18-year-old woman was treated with leukocytapheresis (LCAP) for her combined ulcerative colitis (UC) and aortitis syndrome (AS). Because a close relationship between these two diseases has been suspected based on their etiological and/or pathological findings, we had hypothesized that LCAP, which has satisfactory effects on inflammatory bowel disease such as UC and Crohn's disease might be effective for both her UC and her AS. After informed consent, LCAP therapy was performed once a week for a total of 7 times. Endoscopic remission of the UC was observed. Even though there were no significant improvements in her subjective symptoms of AS such as side-neck pain and dizziness, objective evidence of improvement was obtained when the patient's condition was compared before and after LCAP by angiography, angio-magnetic resonance imaging, and the plethysmogram of her fingertips. These results suggest that LCAP may be valuable as a new adjunct therapy for AS. PMID- 11886586 TI - Trafficking of the Salmonella vacuole in macrophages. AB - Salmonella enterica is a facultative intracellular pathogen which can replicate in macrophages. Intracellular Salmonella exist in a membrane-bound compartment called the Salmonella-containing vacuole. Most studies on Salmonella trafficking in relation to the endocytic pathway have concluded that the majority of Salmonella-containing vacuoles do not interact extensively with late endosomes and lysosomes. Numerous bacterial genes have been identified which are required for survival and replication in macrophages. These include the spv operon, located on the large virulence plasmid, the phoP-phoQ regulon, and those connected with the Salmonella pathogenicity island 2 type III secretion system. The functions of some of these genes are beginning to be understood. In this review, I discuss their roles in relation to our broader understanding of Salmonella trafficking in macrophages. PMID- 11886587 TI - A microbial strategy to multiply in macrophages: the pregnant pause. AB - Humans live in harmony with much of the microbial world, thanks to a sophisticated immune system. As the first line of defense, macrophages engulf, digest, and display foreign material, then recruit specialists to eliminate potential threats. Yet infiltrators exist: certain fungi, viruses, parasites, and bacteria thrive within sentinel macrophages. By scrutinizing the life styles of these shrewd microbes, we can deduce how macrophages routinely mount an effective immune response. The bimorphic life cycles of three pathogens have dramatic consequences for phagosome traffic. In the transmissible state, Leishmania spp., Coxiella burnetii, and Legionella pneumophila block phagosome maturation; after a pregnant pause, replicative forms emerge and thrive in lysosomes. PMID- 11886588 TI - Type IVB secretion by intracellular pathogens. AB - A growing number of pathogens are being found to possess specialized secretion systems which they use in various ways to subvert host defenses. One class, called type IV, are defined as having homology to the conjugal transfer systems of naturally occurring plasmids. It has been proposed that pathogens with type IV secretion systems have acquired and adapted the conjugal transfer systems of plasmids and now use them to export toxins. Several well-characterized intracellular pathogens, including Legionella pneumophila, Coxiella burnetii, Brucella abortus, and Rickettsia prowazekii, contain type IV systems which are known or suspected to be of critical importance in their ability to cause disease. Specifically, these systems are believed to be the key factors determining intracellular fate, and thus the ability to replicate and cause disease. PMID- 11886589 TI - Visualization of actin dynamics during macropinocytosis and exocytosis. AB - Macropinocytosis of newly formed resides and exocytosis of post-lysosomes have been visualized using a green fluorescent protein probe that binds specifically to F-actin filaments. F-actin association with macropinocytosis begins as a V shaped infolding of the membrane. Vesicle enlargement occurs through an inward movement of the proximal point of the V as well as an outward protrusion at the tip of the V to form an elongated invagination. The protrusion eventually closes at its distal margin to become a vesicle and is moved centripetally while recovering its circular shape. The vesicle loses its actin coat within 1 min after internalization. One hour later, post-lysosomal vesicles became weakly surrounded by actin while still cytoplasmic. Some of these vesicles moved to the plasma membrane, docked, and then expelled their contents. Slightly before the vesicle content began to disappear, an increase in F-actin association with the vesicle was observed. This was followed by rapid contraction of the vesicle and then disappearance of the actin signal once the internal content was released. These results show that dynamic changes in actin filament association with the vesicle membrane accompany both endocytosis and exocytosis. PMID- 11886590 TI - The leaden gene product is required with Rab27a to recruit myosin Va to melanosomes in melanocytes. AB - The function of lysosome-related organelles such as melanosomes in melanocytes, and lytic granules in cytotoxic T lymphocytes is disrupted in Griscelli syndrome and related diseases. Griscelli syndrome results from loss of function mutations in either the RAB27A (type 1 Griscelli syndrome) or MYO5A (type 2 Griscelli syndrome) genes. Melanocytes from Griscelli syndrome patients and respective murine models ashen (Rab27a mutant), dilute (myosin Va mutant), and leaden exhibit perinuclear clustering of melanosomes. Recent work suggests that Rab27a is required to recruit myosin Va to melanosomes, thereby tethering melanosomes to the peripheral actin network and promoting melanosome retention at the tips of melanocytic dendrites. Here, we characterize the function of the leaden gene product. We show that Rab27a, but not myosin Va, can be localized to melanosomes in leaden melanocytes, suggesting that the leaden gene product acts downstream of, or in parallel to, Rab27a in melanocytes to promote recruitment of myosin Va to melanosomes. We also observed reduced levels of myosin Va protein in leaden and ashen melanocytes, suggesting that myosin Va stability is influenced by the leaden and ashen gene products. In leaden cytotoxic T lymphocytes, we observed that lytic granules polarize towards the immunological synapse and kill target cells normally. However, in contrast to melanocytes, we found that neither the leaden gene product (melanophilin) nor myosin Va was detectable in cytotoxic T lymphocytes. These results suggest that Rab27a interacts with different classes of effector proteins in melanocytes and cytotoxic T lymphocytes. PMID- 11886591 TI - Ca(2+) and Mg(2+)/ATP independently trigger homotypic membrane fusion in gastric secretory membranes. AB - Exocytic activation of gastric parietal cells represents a massive transformation. We studied a step in this process, homotypic fusion of H,K-ATPase containing tubulovesicles, using R18 dequenching. Ca(2+) and Mg(2+)/ATP each caused dramatic dequenching, reflecting a change in R18 distribution from 5% to 65-90% of the assay's membranes in 2.5 min. These stimuli also triggered fusion between tubulovesicles and liposomes. Independent confirmation that dequenching represented membrane fusion was established by separating tubulovesicle-liposome fusion products on density gradients. Only agents that trigger fusion allowed the transmembrane H,K-ATPase to move to low-density fractions along with R18. EC(50) for Ca(2+)-triggered fusion was 150 nm and for Mg(2+)/ATP-triggered fusion 1 mm, the latter having a Hill coefficient of 2.5. ATP-triggered fusion was specific for Mg(2+)/ATP, required ATP hydrolysis, and was insensitive to inhibition of NSF and/or H,K-ATPase. Fusion initiated by either trigger caused tubulovesicles to become resistant to subsequent challenge by either trigger. Ca(2+) and Mg(2+)/ATP triggered fusion required protein component(s) in tubulovesicles, though this was required in only one of the fusing membranes since tubulovesicles fused well with liposomes containing no proteins. Our data suggest that exocytosis in parietal cells is triggered by separate but interacting pathways and is regulated by self inhibition. PMID- 11886592 TI - Localization of HCMV UL33 and US27 in endocytic compartments and viral membranes. AB - The human cytomegalovirus genome encodes four putative seven transmembrane domain chemokine receptor-like proteins. Although important in viral pathogenesis, little is known about the properties or functions of these proteins. We previously reported that US28 is located in endocytic vesicles and undergoes constitutive endocytosis and recycling. Here we studied the cellular distributions and trafficking of two other human cytomegalovirus chemokine receptor-like proteins, UL33 and US27, in transfected and human cytomegalovirus infected cells. Immunofluorescence staining indicated that UL33 and US27 are located at the cell surface, although the majority of both proteins was seen in intracellular organelles located in the perinuclear region of the cell. The intracellular pools of UL33 and US27 showed overlap with markers for endocytic organelles. Antibody-feeding experiments indicated that cell surface US27 undergoes endocytosis. By immunogold labeling of cryosections and electron microscopy, UL33 was seen to localize to multivesicular bodies (MVBs or multivesicular endosomes). Electron microscopy analysis of human cytomegalovirus infected cells showed that most virus particles wrapped individually into short membrane cisternae, although virus particles were also occasionally seen within and budding into MVBs. Electron microscopy immunolocalization of viral UL33 and US27 on ultrathin cryosections of human cytomegalovirus-infected cells showed gold particles over the membranes into which virions were wrapping, in small membrane tubules and vesicles and in MVBs. Labeling of the human cytomegalovirus glycoproteins gB and gH indicated that these proteins were also present in the same membrane structures. This first electron microscopy analysis of human cytomegalovirus assembly using immunolabeling suggests that the localization of UL33, US27 and US28 to endosomes may allow these proteins to be incorporated into the viral membrane during the final stages of human cytomegalovirus assembly. PMID- 11886593 TI - Meeting report from the EMBO workshop "The Cell Biology of Virus Infection", Heidelberg, Germany, 22-26 September 2001. PMID- 11886595 TI - Molecular cloning of the rat proteinase-activated receptor 4 (PAR4). AB - BACKGROUND: The proteinase-activated receptor 4 (PAR4) is a G-protein-coupled receptor activated by proteases such as thrombin and trypsin. Although activation of PAR4 has been shown to modulate rat gastrointestinal motility, the rat PAR4 sequence was unknown until now. This study aimed to identify the rat PAR4 cDNA. RESULTS: The cDNA coding for the rat PAR4 homologue was cloned from the duodenum. Northern blots demonstrated a 3.0 kb transcript in the duodenum. Protein homology with mouse and human counterparts was 90% and 75% respectively. PAR4 is expressed predominantly in the esophagus, stomach, duodenum and the spleen. When expressed in COS cells, PAR4 is activated by trypsin (1 nM), thrombin (50 nM), mouse PAR4 specific peptide (500 microM) and a putative rat PAR4 specific activating peptide (100 microM), as measured by intracellular Ca2+-changes. CONCLUSIONS: We have identified and characterized cDNA encoding the rat PAR4 homologue. PAR4 is expressed predominantly in the upper gastrointestinal tract. It is activated by trypsin, thrombin and its newly identified rat PAR4 specific activating peptide. PMID- 11886596 TI - Gender assignment and ideology. PMID- 11886597 TI - Clinical and biochemical outcome of conventional dose radiotherapy for localized prostate cancer. AB - PURPOSE: To retrospectively review the results of conventional dose radical radiotherapy for clinical stage T1 and T2 prostate cancer, and to identify the factors that predict the biochemical relapse-free rate. METHODS: The records were reviewed of 706 hormonally-naive men with clinical stage T1T2 prostate cancer treated with radical radiotherapy (RT) between 1987-1994 at the Princess Margaret Hospital. The median prostate RT dose was 65 Gy in 35 fraction (range 52 Gy in 20 fractions to 67 Gy in 37 fractions). Pelvic lymph nodes were included in the treatment volume and treated to a median dose of 45 Gy in 25 fractions for 283 cases (40%). The primary end-point was biochemical relapse-free survival from RT using the American Society of Therapeutic Radiology and Oncology (ASTRO) consensus criteria. Favourable, intermediate and unfavourable pre- treatment prognostic groupings were derived from the initial PSA, T-category, and Gleason score using Cox regression analysis. Secondary end-points included survival, metastases-free survival and clinical local control. RESULTS: The overall biochemical relapse-free rate at 2 and 5 years was 63% and 45% respectively. Overall survival at 5 years was 87%, and metastases-free survival was 86%. Local control by DRE was 72% at 5 years. Multivariate analysis of variables associated with time to biochemical failure after RT showed that pre-RT PSA, T-category and Gleason score were significant independent predictors with hazard ratios of 1.33 (P = 0.0001), 1.22 (P = 0.01) and 1.33 (P = 0.029) respectively. PSA nadir was an early indicator of biochemical failure. The biochemical failure rate at 3 years was 20% for a PSA nadir < or = 0.5 ng/ml and 85% for a PSA nadir > or = 2.0 ng/ml (P < 0.0001). CONCLUSION: The results of conventional dose RT were unsatisfactory for all risk categories, and overall, less than half of the treated patients remained in biochemical remission at 5 years. These men require more aggressive therapy, that may include dose escalation with conformal techniques, and neoadjuvant/adjuvant androgen deprivation therapy. These results highlight the need to support new and on-going clinical trials for management of localized disease. PMID- 11886599 TI - The efficacy of chondroitin sulfate 0.2% in treating interstitial cystitis. AB - OBJECTIVE: An open label study of chondroitin sulfate was undertaken to determine the response of patients with interstitial cystitis and positive potassium test results to this agent. METHOD: Eighteen patients with classic features of interstitial cystitis were enrolled in the study. Patients received 40 mL chondroitin sulfate, 0.2% instilled intravesically once a week for four weeks and then once a month for 12 months. At the same times, Quality of Life Improvement scores, voiding diaries, and pain and voiding indices were reviewed. RESULTS: Thirteen of 18 patients were followed for the entire 13-month study. Twelve of these patients responded to treatment within 3 to 12 weeks, on average. A total of 6/13 (46.2%) showed a good response, 2/13 (15.4%) had a fair response, and 4/13 (30.8%) had a partial response and 1/13 (7.7%) showed no response. CONCLUSION: Intravesical chondroitin sulfate seems to demonstrate some beneficial effects in treatment of interstitial cystitis patients who have positive potassium stimulation test results. PMID- 11886600 TI - Greater reliability of neonatal ultrasonography in defining renal hypoplasia with antenatal hydronephrosis and vesicoureteral reflux. AB - PURPOSE: Infants with history of antenatal hydronephrosis and neonatal vesicoureteral reflux may have detectable changes in renal scans before the advent of urinary tract infection. In cases of bilateral high-grade vesicoureteral reflux, differential renal function on renal scan may not reveal renal hypoplasia since comparison of relative function may be made between two abnormal kidneys. We tested the hypothesis that ultrasonography in the neonatal period may be accurate and complementary to renal scan in detecting renal hypoplasia at birth. MATERIALS AND METHODS: Twenty-six infants who presented in the antenatal period with history of hydronephrosis and were noted to have neonatal vesicoureteral reflux postnatally were studied retrospectively. They had all been treated by a prospective protocol that included renal ultrasound and renal scans in the first 6 weeks of life. All had been placed on prophylactic antibiotics and had no urinary tract infection. Multiple sonographic parameters were analyzed including kidney length, echogenicity, calyceal blunting, parenchymal thinning and focal scars. We correlated the renal morphology on ultrasound, the renal function on renal scan and the degree of reflux seen on VCUG. RESULTS: VCUG showed reflux in 44 renal units, grade of reflux was: I (2), II (7), III (12), IV (8), and V (15). A variety of nucleides were used including DMSA in 15, DTPA in 6 and MAGIII in 5. Renal scans identified global hypoplasia without focal scars (differential function less than 40%) in 10 of 44 refluxing renal units grades I (1), III (2), IV (4), and V (3). The sonographic finding of decreased renal length (<50th percentile for age) was present in 14 refluxing units of 44 refluxing renal units, grade I (1), grade III (4 bilaterally in 1), grade IV (4 bilaterally in 1), and grade V (5 bilaterally in 2). The sonographic finding of decreased renal length (<50th percentile) correlated strongly with renal hypoplasia on renal scans in refluxing renal units (p value <.005, sensitivity 80% and specificity 82%, positive predictive value 57%, and negative predictive value 93%). CONCLUSION: Postnatal ultrasonography is a reliable measure of gross renal parenchyma, and in the presence of vesicoureteral reflux correlates with renal scintilligraphy. In addition, for cases of bilateral neonatal vesicoureteral reflux, ultrasound and renal scan are complimentary, each being able to detect the abnormalities that might be missed by the other. PMID- 11886601 TI - Sclerosing lipogranuloma: an unusual scrotal mass. AB - Sclerosing lipogranuloma of the male genitalia without a history of injection of exogenous material is extremely rare. This is the first case reported from a Canadian center. This 33 year old man developed sclerosing lipogranuloma of his scrotum 3 months after being diagnosed with infectious mononucleosis. There was no history of injection of exogenous substances or trauma. His lesion was painless, sudden in onset, "Y-shaped", associated with eosinophila and spontaneously regressed after partial resection. A review of the available English literature on sclerosing lipogranuloma from 1966 to 2001 was completed to compare our case report to previously available reports. The results show definite differences in the presentation of primary versus secondary sclerosing lipogranuloma. Sixty-eight per cent of the cases of primary sclerosing lipogranuloma involved the scrotum only while 63% of secondary sclerosing lipogranuloma involved the penis only. Seven per cent of lesions attributed to primary sclerosing lipogranuloma were painful compared to 69% of secondary sclerosing lipogranulomas. Cases of primary sclerosing lipogranuloma were often described as "Y-shaped" and were unlikely to recur. Understanding the typical presentation of this condition will allow future cases to be recognized more easily and managed appropriately. Primary sclerosing lipogranuloma may be diagnosed by fine needle aspiration or excisional biopsy and then managed conservatively avoiding more complex and invasive surgery. PMID- 11886602 TI - Palliative Subcutaneous Tunneled Nephrostomy Tube (PSTN): a simple and effective technique for management of malignant extrinsic ureteral obstruction. AB - The establishment and maintenance of effective urinary tract drainage for patients with malignant extrinsic ureteric obstruction is a formidable challenge for the urologist. We have utilized an alternative method of urinary diversion, called Palliative Subcutaneous Tunneled Nephrostomy Tubes (PSTN), for long term urinary tract drainage when intracoropreal stenting has failed or is not tolerated. PSTN provides a simple and effective method of external urinary diversion and preservation of renal function. This technique should be an option in the armamentarium of urologists for management of malignant ureteral obstruction. PMID- 11886603 TI - Genital pain of unknown aetiology. PMID- 11886604 TI - Treatment of genital warts - what's the evidence? AB - Genital warts are usually asymptomatic, and rarely cause discomfort. Once the patient is aware of them the main symptom is their cosmetic appearance and resultant psychological consequences. The ideal treatment outcome would be complete viral eradication, but this is not possible. Treatments focus on the removal of exophytic warts, leaving the surrounding subclinical and latent human papillomavirus (HPV) infection as areas of possible transmission and recurrence. Effective treatment does reduce HPV viral load, so the infection is reduced if not completely eradicated. Treatment is often painful, inconvenient, and may produce poor clearance rates and frequent recurrences. The treatment chosen should be no worse or more dangerous than the disease itself, and should be tailored to the patients' disease and needs as well as to the available resources. Genital warts are highly infectious and sexual partners may well already be infected when a patient presents for treatment. There are no published studies showing that condom use reduces transmission of HPV from people with genital warts. However, if the sexual partner is uninfected; using a condom may protect against HPV lesions and genital warts. Condom use should be encouraged in new relationships. PMID- 11886605 TI - Information-giving to patients with genital warts at a genitourinary medicine clinic: a baseline assessment. AB - Aspects of information-giving to patients with a first episode of genital warts attending a large UK genitourinary medicine clinic on three or more occasions were assessed using a questionnaire survey employing correlational and between groups analyses. The main outcomes measured were levels of, and interrelationships between, patients' perceptions of information received about genital warts, accuracy of knowledge, quality of interaction with clinic personnel, written information, patients' level of anxiety and demographic variables. Patients with genital warts have good basic knowledge about this condition, although understanding about several complex issues is poor. Educational level was positively correlated with accuracy of knowledge about genital warts. Although ease of communicative interaction with information-givers was positively correlated with perceived information given, no relationship was found between the latter and actual knowledge about genital warts. Anxiety levels and receiving written information were largely uncorrelated with accuracy of knowledge about genital warts. This study suggests that while patients' perception of information received may relate to more satisfactory interaction with information-givers, actual knowledge may be a more important measurable outcome. More interventional work is needed to determine how the information giving process for genital warts can be optimized. PMID- 11886606 TI - Sexual health provision and the Sexual Health and HIV Strategy for England: a questionnaire survey of general practitioners' views. AB - An anonymous postal survey was used to assess the views of general practitioners (GPs) on the expected contents of the Department of Health National Sexual Health and HIV Strategy for England prior to release. One hundred and seventeen GPs in four London Primary Care Groups took part (response rate=50%). The majority felt they currently have about the right amount of involvement in patient care (HIV 70%: GUM 66%); 88% of GPs felt they offer general sexual health care (Level one), and most wished to continue with this (85%). However, a sizeable minority of GPs wanted to be involved in the development of care guidelines (HIV 17%; GUM 22%), and 10% considered that they might wish to offer more specialist services (Level two). To achieve a Strategy goal to integrate sexual healthcare services in England, it is likely that there will need to be local consultation and support for GPs. PMID- 11886607 TI - Chlamydial infection in males and conseqences for their female sexual partners, an example from rural Kilimanjaro, Tanzania. AB - The aim of this paper is to examine the reproductive health status for the wives of chlamydia-infected, but largely asymptomatic men. In a cross-sectional study in rural Tanzania 447 men and 393 women, aged 15-44 years, were screened for chlamydial infection. The prevalence was 9.6% and 6.9%, respectively. Among 43 chlamydia-positive men, 17 were married. Data from both spouses, independently examined, could be matched for 12 couples. None of the 12 husbands had discharge, one had dysuria and 3 had pyuria. Three wives tested positive for chlamydial infection, two others had pelvic inflammatory disease, four others had pyuria. Men reported more sexual partners than women and were considered being the index case for the chlamydial infection. While three of 11 wives (27%) tested chlamydia positive from a cervical sample, transmission might have occurred in 5-8 cases (transmission rate (42-67%), indicating that these 12 largely asymptomatic chlamydia-positive men were highly infectious to their sexual partners. PMID- 11886608 TI - Risk behaviours and prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae genital infections among Montreal street youth. AB - We estimated the prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae genital tract infections among 302 Montreal street youth (223 boys) and identified associated risk factors. Study participants, 14-25 years old (average 20.9 years), meeting specific criteria for homelessness, were recruited in street youth agencies. Participation included a structured interview and provision of a urine specimen. Among sexually active youth, (n = 300) 30.0% had more than five heterosexual partners and 13.0% had at least one homosexual partner (last year), 10.7% had received money in exchange for sex (last six months) and 47.0% reported sexual relations resulting in pregnancy (lifetime). Among all youths, 82.1% had used at least one type of illicit drug, and 30.1% injected drugs at least once (last six months). The prevalence of C. trachomatis infection was 6.6% (95% CI 4.1-10.0%). Prevalence did not vary significantly by sex, age or any other variable, except history of pregnancy (10.4% among youth with history of pregnancy vs 3.6% among others, P = 0.02). No cases of N. gonorrhoeae infection were found. PMID- 11886609 TI - Sexually transmitted diseases in Germany. AB - In the former West Germany, in specific venereal diseases legislation passed in 1953, only syphilis, gonorrhoea, ulcus molle, and lymphogranuloma venereum were defined as venereal diseases and subject to mandatory notification. The proportion of unreported cases was as high as 75% for syphilis and up to 90% for gonorrhoea. Epidemiological data for the past 10 years exist only on selected populations from research studies and are summarized in this article. In the former East Germany reporting of sexually transmitted infections (STIs) was mandatory and, due to the centralized organization, underreporting was considered to be low, although no specific studies have examined this. After the unification in 1990 of the two German states the West German laws were adopted in East Germany. Since 1982 - when the first AIDS case was reported in Germany - information on AIDS cases has voluntarily been collected at the national register at the AIDS Centre of the Robert Koch Institute in Berlin. The law governing the reporting of infectious diseases has recently been revised. Under the new Protection against Infection Act, which became effective on 1 January 2001, clinical diagnoses of STIs (with the exception of hepatitis B) are no longer notifiable diseases. Laboratory reporting of positive test results for Treponema pallidum has been introduced. With T. pallidum and HIV notifications, additional disaggregated data are collected. Since T. pallidum and HIV remain the only notifiable STIs, all other STIs have to be monitored through sentinel surveillance systems. These surveillance systems are currently being established. Under the new legislation, local health authorities have to provide adequate counselling and testing services for STIs, which may be provided free of charge if necessary. PMID- 11886610 TI - The organization of STI control in the Netherlands - an overview. AB - The organization of STI-control in the Netherlands is described. STIs represent a relatively modest health problem in the Netherlands. Chlamydia trachomatis is the most frequent STI. Recent reports show that STIs are on the rise again, especially amongst gay men. The general health system deals with STIs, the main professionals are: the general practitioners, public health nurses and dermatovenereologists. The larger cities have specific STI-clinics. Prevention is a cornerstone in Dutch STI-control. At a regional level, the municipal health services play a major role in policy and prevention. At a national level, several specific organizations are active. In general the STI-system is of good quality. Future attention is needed for the financial arrangement, surveillance and the rising incidence of STIs. PMID- 11886611 TI - Chlamydial co-infection among patients with gonorrhoea. AB - The objective was to determine how often gonococcal (GC) infection is accompanied by chlamydial co-infection and to determine risk factors for dual infection. All GC-positive cultures were identified between 24 April and 9 September 1998, among patients seen at the three genitourinary medicine (GUM) clinics across the Chelsea and Westminster Directorate. Chlamydia trachomatis was diagnosed using an enzyme-linked immunosorbent assay (ELISA) (Dade-Behring). One hundred and fifty three episodes of gonorrhoea were identified. Information on chlamydial infection was available for 149 cases of GC of whom 16 (10.7%) were found to be co-infected with C. trachomatis. In univariate analysis, chlamydial co-infection was exclusively diagnosed in heterosexuals, and was more likely to be diagnosed among females, in younger individuals and in individuals of black Caribbean ethnic group. In multivariable analyses, however, only the sex and age of the individual were independently associated with chlamydial co-infection. The rate of co infection was 10.7%. Independent risk factors were being less than 20 years old and being female. PMID- 11886612 TI - A multidistrict audit of the management of chlamydial PID in genitourinary medicine clinics in Yorkshire. AB - In response to recent reviews of practice of pelvic inflammatory disease (PID) management, a multidistrict audit involving eight genitourinary clinics within the Yorkshire region was carried out. This audit reports the referral patterns of patients, physical signs and microscopy findings at the first genitourinary medicine (GUM) clinic attendance, antibiotic treatment and follow-up data as well as health adviser involvement and partner participation for 68 patients diagnosed with chlamydial PID. Twenty-eight (41.2%) patients presented with symptoms of less than or equal to four weeks duration, partner notification was recorded as being carried out for all patients and at least one partner was documented as having been treated in 57 (83.8%) patients. There is, however, a wide variation in the antibiotic regimens used for treatment and their duration. PMID- 11886614 TI - The male genital skin burning syndrome (Dysaesthetic Peno/Scroto-dynia). AB - Men may complain of penile and/or scrotal skin burning with no evidence of positive physical signs or investigations. The condition is cumbersome and leads to stress and disruption in social and sexual relationships. The patients report no response to previous medications (including antibiotics, antifungals and topical corticosteroids); and identify improvement in symptoms and quality of life on selective serotonin re-uptake inhibitors (SSRI). A similar condition has been recognized in the female patients (dysaesthetic vulvodynia). We report the occurrence of this condition in three men and suggest it being recognized as 'the male genital skin burning syndrome' (Dysaesthetic Peno/Scroto-dynia). PMID- 11886613 TI - Audit of hepatitis B immunization at the genitourinary medicine department in Middlesbrough, UK. AB - The outcomes of hepatitis B immunization programme at the genitourinary medicine (GUM) department in Middlesbrough during a period of five years were examined. The majority of those immunized were male homosexuals who had earlier requested an HIV test. All participants had preimmunization hepatitis B serology, and were negative. The default rate was high at 20/42 (47.6%). However, among those who completed all the processes, seroconversion rate was high at 21/22 (95.5%). The only non-responder was a man who was HIV-positive. The type of vaccine and immunization regimen are discussed. PMID- 11886615 TI - Subcutaneous granulomatous lesions related to ritonavir therapy in a HIV infected patient. AB - We report a case of subcutaneous granulomatous lesions developed in a HIV seropositive man, related to ritonavir therapy. The importance of close monitoring and investigations of patients developing unusual side effects during highly active antiretroviral therapy (HAART) is demonstrated. PMID- 11886616 TI - To cut a long story short - the use of imiquimod in resistant warts. AB - Persistent anogenital warts are frustrating and expensive to manage, both for the patient and the clinic. This case report illustrates prolonged and varied management, and eventual success with imiquimod. PMID- 11886617 TI - Donovanosis in India: declining fast? PMID- 11886619 TI - Generation of periodic waves by landscape features in cyclic predator-prey systems. AB - The vast majority of models for spatial dynamics of natural populations assume a homogeneous physical environment. However, in practice, dispersing organisms may encounter landscape features that significantly inhibit their movement. We use mathematical modelling to investigate the effect of such landscape features on cyclic predator-prey populations. We show that when appropriate boundary conditions are applied at the edge of the obstacle, a pattern of periodic travelling waves develops, moving out and away from the obstacle. Depending on the assumptions of the model, these waves can take the form of roughly circular 'target patterns' or spirals. This is, to our knowledge, a new mechanism for periodic-wave generation in ecological systems and our results suggest that it may apply quite generally not only to cyclic predator-prey interactions, but also to populations that oscillate for other reasons. In particular, we suggest that it may provide an explanation for the observed pattern of travelling waves in the densities of field voles (Microtus agrestis) in Kielder Forest (Scotland-England border) and of red grouse (Lagopus lagopus scoticus) on Kerloch Moor (northeast Scotland), which in both cases move orthogonally to any large-scale obstacles to movement. Moreover, given that such obstacles to movement are the rule rather than the exception in real-world environments, our results suggest that complex spatio-temporal patterns such as periodic travelling waves are likely to be much more common in the natural world than has previously been assumed. PMID- 11886620 TI - Understanding the persistence of measles: reconciling theory, simulation and observation. AB - Ever since the pattern of localized extinction associated with measles was discovered by Bartlett in 1957, many models have been developed in an attempt to reproduce this phenomenon. Recently, the use of constant infectious and incubation periods, rather than the more convenient exponential forms, has been presented as a simple means of obtaining realistic persistence levels. However, this result appears at odds with rigorous mathematical theory; here we reconcile these differences. Using a deterministic approach, we parameterize a variety of models to fit the observed biennial attractor, thus determining the level of seasonality by the choice of model. We can then compare fairly the persistence of the stochastic versions of these models, using the 'best-fit' parameters. Finally, we consider the differences between the observed fade-out pattern and the more theoretically appealing 'first passage time'. PMID- 11886621 TI - Mitochondrial gene rearrangements confirm the parallel evolution of the crab-like form. AB - The repeated appearance of strikingly similar crab-like forms in independent decapod crustacean lineages represents a remarkable case of parallel evolution. Uncertainty surrounding the phylogenetic relationships among crab-like lineages has hampered evolutionary studies. As is often the case, aligned DNA sequences by themselves were unable to fully resolve these relationships. Four nested mitochondrial gene rearrangements--including one of the few reported movements of an arthropod protein-coding gene--are congruent with the DNA phylogeny and help to resolve a crucial node. A phylogenetic analysis of DNA sequences, and gene rearrangements, supported five independent origins of the crab-like form, and suggests that the evolution of the crab-like form may be irreversible. This result supports the utility of mitochondrial gene rearrangements in phylogenetic reconstruction. PMID- 11886622 TI - Evidence for habitat partitioning based on adaptation to environmental light in a pair of sympatric lizard species. AB - Terrestrial habitats exhibit a variety of light environments. If species exhibit evolutionary adaptations of their visual system or signals to habitat light conditions, then these conditions can directly influence the structure of communities. We evaluated habitat light characteristics and visual-signal design in a pair of sympatric species of lizards: Anolis cooki and Anolis cristatellus. We found that each species occupies a distinct microhabitat with respect to light intensity and spectral quality. We measured the relative retinal spectral sensitivity and found significant differences between the species that correlate with differences in habitat spectral quality. We measured the spectral reflectance of the dewlaps (colourful throat fans used in communication), and found that the A. cooki dewlap reflects little ultraviolet (UV), while that of A. cristatellus reflects strongly in the UV. For both species downwelling light (irradiance) is rich in UV. However the background light (radiance) is rich in UV for A. cooki, but low in UV for A. cristatellus. Thus, the dewlap of each species creates a high contrast with the background in the UV. Our findings strongly suggest that these two species are partitioning their habitat through specializations of the visual system and signal design to microhabitat light conditions. PMID- 11886623 TI - Polyandry produces sexy sons at the cost of daughters in red flour beetles. AB - Female mating with multiple males within a single fertile period is a common phenomenon in the animal kingdom. Female insects are particularly promiscuous. It is not clear why females mate with multiple partners despite several potential costs, such as expenditure of time and energy, reduced lifespan, risk of predation and contracting sexually transmitted diseases. Female red flour beetles (Tribolium castaneum) obtain sufficient sperm from a single insemination to retain fertility for several months. Nonetheless they copulate repeatedly within minutes with different males despite no direct fitness benefits from this behaviour. One hypothesis is that females mate with multiple partners to provide indirect benefits via enhanced offspring fitness. To test this hypothesis, we compared the relative fitness of F(1) offspring from females mated with single males and multiple males (2, 4, 8, or 16 partners), under the condition of relatively high intraspecific competition. We found that a female mating with 16 males enhanced the relative fitness of F(1) males (in two out of three trials) but reduced F(1) females' fitness (in two independent trials) in comparison with singly mated females. We also determined whether several important fitness correlates were affected by polyandry. We found that F(1) males from mothers with 16 partners inseminated more females than F(1) males from mothers with a single partner. The viability of the eggs sired or produced by F(1) males and females from highly polyandrous mothers was also increased under conditions of low intra specific competition. Thus, the effects of polyandry on F(1) offspring fitness depend on environmental conditions. Our results demonstrated a fitness trade-off between male and female offspring from polyandrous mothers in a competitive environment. The mechanisms and biological significance of this unique phenomenon are discussed. PMID- 11886624 TI - Origin of the tooth-replacement pattern in therian mammals: evidence from a 110 Myr old fossil. AB - Living placental and marsupial mammals (therians) use distinctive tooth replacement patterns that have not yet been traced back fully to their time of divergence in the Early Cretaceous (>100 Myr ago). Slaughteria eruptens, a small 110 Myr old fossil mammal from Texas, USA, is near the base of that divergence. Using ultra-high-resolution X-ray CT analysis we demonstrate that Slaughteria preserves an unrecognized pattern of tooth replacement with simple posterior premolars replacing molariform precursors. Differing from both placentals that have a more complex posterior adult premolar, and from marsupials, in which only one premolar is replaced, Slaughteria provides the first direct evidence of a tooth-replacement pattern that is plausible for the common ancestor of all therians. By our interpretation Slaughteria has only one adult molar in place and contains two mental foramina in the jaw, thus changing characters that are critical to reconstruction of mammalian relationships and to species discrimination and interpretations of diversity for Early Cretaceous mammals. PMID- 11886625 TI - The evolution of cuckoo parasitism: a comparative analysis. AB - Cuckoos (family Cuculidae) show the highest diversity of breeding strategies within one bird family (parental care, facultative and obligate brood parasites). We used independent contrasts from two phylogenies to examine how this variation was related to 13 ecological and life-history variables. The ancestral state was probably tropical, resident, forest cuckoos with parental care. The evolution of brood parasitism was correlated with a shift to more open habitats, a change in diet, increases in species breeding-range size and migration, and a decrease in egg size. Once parasitism had evolved, more elaborate parasitic strategies (more harmful to host fitness) were correlated with decreased egg size, a change in diet, increased breeding-range size and migration, a shortened breeding season and a decrease in local abundance. Establishing the most probable evolutionary pathways, using the method of Pagel, shows that changes in ecological variables (such as migration, range size and diet type) preceded the evolution of brood parasitism, which is likely to be a later adaptation to reduce the cost of reproduction. By contrast, brood parasitism evolved before changes in egg size occurred, indicating that egg size is an adaptive trait in host--parasite coevolution. Our results suggest that the evolution of cuckoo brood parasitism reflects selection from both ecological pressures and host defences. PMID- 11886626 TI - Use of population genetic data to infer oviposition behaviour: species-specific patterns in four oak gallwasps (Hymenoptera: Cynipidae). AB - Many species of oak gallwasp (Hymenoptera: Cynipidae: Cynipini) induce galls containing more than one larva (multilocular galls) on their host plant. To date, it has remained unclear whether multilocular galls result solely from clustered oviposition by a single female, or include the aggregated offspring of several females (multiple founding). We have developed a novel maximum-likelihood approach for use with population genetic data that estimates the number and genotypes of parents contributing to offspring from each gall. We apply this method to allozyme data from multiple populations of four oak gallwasps whose asexual generations develop in multilocular galls (Andricus coriarius, A. lucidus, A. panteli and A. seckendorffi). We find strong evidence for multiple founding in all four species, and show the data to be compatible with multiple founding rather than founding by a single foundress mated with multiple males. The extent of multiple founding differs among species: in A. lucidus and A. seckendorffi most galls are induced by a single female, whereas in A. coriarius and A. panteli over half of the galls sampled were multiple founded. We suggest that variation in levels of multiple founding may be due to consistent ecological differences between the four species. PMID- 11886627 TI - North Atlantic Oscillation synchronizes food-web interactions in central European lakes. AB - A regular and distinct feature of seasonal plankton succession in temperate lakes is the early summer period of algal suppression by herbivores, i.e. the clear water phase. Within the last 30 years the timing of this food-web interaction between algae and herbivores has advanced on average by approximately two weeks in central European lakes due to faster population growth of herbivores in warmer water. Trend and inter-annual variability in clear water timing were strongly related to the climate dynamics of the North Atlantic, i.e. the North Atlantic Oscillation (NAO). Due to its large-scale effects, the NAO synchronized plankton succession in central European lakes, causing a striking temporal coherence of a food-web interaction over several hundreds of kilometers. PMID- 11886628 TI - Mixed inoculation alters infection success of strains of the endophyte Epichloe bromicola on its grass host Bromus erectus. AB - Within-host competition in multiply infected hosts is considered an important component of host-parasite interactions, but experimental studies on the dynamics of multiple infections are still rare. We measured the infection frequencies of four strains of the fungal endophyte Epichloe bromicola on two genotypes of its host plant Bromus erectus after single- and double-strain inoculation. Double strain inoculations resulted in fewer double, but more single, infections than expected on the basis of infection frequencies in single-strain inoculations. In most cases, only one of the two strains established an infection, and strains differed in their overall competitive ability. This pattern resembles the mutual exclusion scenarios in some theoretical models of parasite evolution. In addition, competitive ability varied with host genotype, which may represent a mechanism for the coexistence of strains in a population. Hence, considering the genetic variation in both host and parasite may be important for a better understanding of within-host dynamics and their role in epidemiology or (co)evolution. PMID- 11886629 TI - State-dependent behaviour in breeding barn swallows (Hirundo rustica): consequences for reproductive effort. AB - Life-history theory offers an explanation for the intraspecific variation in reproductive effort; increased levels of current reproductive success, for example, may trade off against residual reproductive value. Even where such trade offs have been demonstrated, however, much variation in effort remains unexplained and the underlying causes are usually obscure. We examined body state, i.e. energy reserves, as a factor, which could moderate reproductive effort. Specifically, overnight heating and cooling treatments were used to adjust dawn energy reserves in female swallows attending their nests without impinging on the opportunities for foraging. Changes in reproductive effort were measured as 'daytime energy expenditure' (doubly labelled water technique) and the 'number of feeding visits' during brood rearing, which both relate positively to current reproductive success. Our experimental treatments and responses were then compared using the common currency of energy. In response to positive and negative state manipulations, female swallows increased and decreased, respectively, their daytime energy expenditure (and number of feeding visits). These responses to experimental manipulation of state provided evidence of a direct link between the energy expenditure, life history and behaviour, which has hitherto proved elusive. They allow that energy supply and expenditure play a regulatory role in reproductive effort, and indicate that units of energy expenditure probably carry fitness costs and benefits, which are context dependent. PMID- 11886630 TI - The relationship between mimetic imperfection and phenotypic variation in insect colour patterns. AB - Many hoverflies (Syrphidae) mimic wasps or bees through colour or behavioural adaptations. The relationship between phenotypic variation in colour pattern and mimetic perfection (as determined by pigeons) was investigated in three species of Mullerian mimics (Vespula spp.) and 10 Batesian hoverfly mimics, plus two non mimetic species of flies. Four predictions were tested: (i) Batesian mimics might be imperfect because they are in the process of evolving towards perfection, hence there should be a positive relationship between variation and imperfection; (ii) some Batesian mimics are imperfect because they do not have the appropriate genetic variation to improve and have evolved to be as good as possible, hence there should be no differences between species, all displaying a low level of variation; (iii) very common hoverflies should show the highest levels of variation because they outnumber their models, resulting in high predation and a breakdown in the mimetic relationship; and (iv) social wasps (Vespula) have such a powerful defence that anything resembling a wasp, both Mullerian and perfect Batesian mimics, would be avoided, resulting in relaxed selection and high variance. Poor mimics may still evolve to resemble wasps as well as possible and display lower levels of variation. The data only provided support for the fourth prediction. The Mullerian mimics, one of the most perfect Batesian mimics, and the non-mimetic flies displayed much higher levels of variation than the other species of Batesian mimics. PMID- 11886631 TI - Adaptive production of fighter males: queens of the ant Cardiocondyla adjust the sex ratio under local mate competition. AB - Hamilton's concept of local mate competition (LMC) is the standard model to explain female-biased sex ratios in solitary Hymenoptera. In social Hymenoptera, however, LMC has remained controversial, mainly because manipulation of sex allocation by workers in response to relatedness asymmetries is an additional powerful mechanism of female bias. Furthermore, the predominant mating systems in the social insects are thought to make LMC unlikely. Nevertheless, several species exist in which dispersal of males is limited and mating occurs in the nest. Some of these species, such as the ant Cardiocondyla obscurior, have evolved dimorphic males, with one morph being specialized for dispersal and the other for fighting with nest-mate males over access to females. Such life history, combining sociality and alternative reproductive tactics in males, provides a unique opportunity to test the power of LMC as a selective force leading to female-biased sex ratios in social Hymenoptera. We show that, in concordance with LMC predictions, an experimental increase in queen number leads to a shift in sex allocation in favour of non-dispersing males, but does not influence the proportion of disperser males. Furthermore, we can assign this change in sex allocation at the colony level to the queens and rule out worker manipulation. PMID- 11886632 TI - Parental care of a cowbird host: caught between the costs of egg-removal and nest predation. AB - Avian brood parasites reduce host fitness through the addition of parasitic eggs and the removal of host eggs. Both parasitic egg-addition and host egg-removal may be important sources of selection on host behaviour, creating fitness trade offs with selection imposed by nest predation. However, the relative costs hosts suffer from egg-addition and host egg-removal and the responses to these costs are largely unstudied. Through experimental manipulations and observations, we demonstrate that increased nest attentiveness by female yellow warblers (Dendroica petechia) reduces the cost of brood parasitism by reducing egg-removal by brown-headed cowbirds (Molothrus ater). However, female attentiveness does not reduce the addition of parasitic eggs. Experimentally parasitized females respond to the threat of egg-removal by increasing nest attentiveness. Increased attentiveness, however, reduces time for females to gather food and requires males to visit the nest more often to feed incubating females. This increased activity in turn increases the risk of nest predation. Thus, brood parasitism (specifically egg-removal) and nest predation produce conflicting selection on incubation strategies, as parasitized hosts are caught between the costs of egg removal by brood parasites, and the costs of increased nest predation if the female spends more time on the nest to reduce egg-removal. PMID- 11886633 TI - A large gene family for putative variant antigens shared by human and rodent malaria parasites. AB - A major mechanism whereby malaria parasites evade the host immune response to give chronic infections in patients' blood for months, or even years, is antigenic variation. In order to generate variant antigens, parasites require large multigene families. Although several gene families involved in these phenomena have been identified in the human malaria Plasmodium falciparum, to date no variant antigen gene families have been identified in malaria species that will infect widely used rodent laboratory hosts. Here we present, for the first time, to our knowledge, a large multigene family conserved in both rodent and human malarias, which is a strong candidate as a major variant antigen gene family. In each of four species of Plasmodium, three rodent malarias and the human pathogen P. vivax, homologues of the gene family were found to have a conserved three-exon structure. In the rodent malaria P. chabaudi, transcription of members of the gene family was developmentally regulated with maximum expression in late trophozoite stages, which is the developmental stage known to express variant antigen proteins. PMID- 11886634 TI - The effect of Wolbachia-induced cytoplasmic incompatibility on host population size in natural and manipulated systems. AB - Obligate, intracellular bacteria of the genus Wolbachia often behave as reproductive parasites by manipulating host reproduction to enhance their vertical transmission. One of these reproductive manipulations, cytoplasmic incompatibility, causes a reduction in egg-hatch rate in crosses between individuals with differing infections. Applied strategies based upon cytoplasmic incompatibility have been proposed for both the suppression and replacement of host populations. As Wolbachia infections occur within a broad range of invertebrates, these strategies are potentially applicable to a variety of medically and economically important insects. Here, we examine the interaction between Wolbachia infection frequency and host population size. We use a model to describe natural invasions of Wolbachia infections, artificial releases of infected hosts and releases of sterile males, as part of a traditional sterile insect technique programme. Model simulations demonstrate the importance of understanding the reproductive rate and intraspecific competition type of the targeted population, showing that releases of sterile or incompatible individuals may cause an undesired increase in the adult number. In addition, the model suggests a novel applied strategy that employs Wolbachia infections to suppress host populations. Releases of Wolbachia-infected hosts can be used to sustain artificially an unstable coexistence of multiple incompatible infections within a host population, allowing the host population size to be reduced, maintained at low levels, or eliminated. PMID- 11886635 TI - Short day lengths attenuate the symptoms of infection in Siberian hamsters. AB - Symptoms of infection, such as fever, anorexia and lethargy, are ubiquitous among vertebrates. Rather than nonspecific manifestations of illness, these responses are organized, adaptive strategies that are often critical to host survival. During times of energetic shortage such as winter, however, it may be detrimental for individuals to prolong energetically demanding symptoms such as fever. Individuals may adjust their immune responses prior to winter by using day length to anticipate energetically-demanding conditions. If the expression of sickness behaviours is constrained by energy availability, then cytokine production, fever, and anorexia should be attenuated in infected Siberian hamsters housed under simulated winter photoperiods. We housed hamsters in either long (14 L : 10 D) or short (10 L : 14 D) day lengths and assessed cytokines, anorexia and fever following injections of lipopolysaccharide (LPS). Short days attenuated the response to lipopolysaccharide, by decreasing the production of interleukin (IL) 6 and IL-1beta, and diminishing the duration of fever and anorexia. Short-day exposure in hamsters also decreased the ingestion of dietary iron, a nutrient vital to bacterial replication. Taken together, short day lengths attenuated the symptoms of infection, presumably to optimize energy expenditure and survival outcome. PMID- 11886636 TI - Volatiles from potato plants infected with potato leafroll virus attract and arrest the virus vector, Myzus persicae (Homoptera: Aphididae). AB - The influence of viral disease symptoms on the behaviour of virus vectors has implications for disease epidemiology. Here we show that previously reported preferential colonization of potatoes infected by potato leafroll virus (genus Polerovirus) (luteovirus) (PLRV) by alatae of Myzus persicae, the principal aphid vector of PLRV, is influenced by volatile emissions from PLRV-infected plants. First, in our bioassays both differential immigration and emigration were involved in preferential colonization by aphids of PLRV-infected plants. Second, M. persicae apterae aggregated preferentially, on screening above leaflets of PLRV-infected potatoes as compared with leaflets from uninfected plants, or from plants infected with potato virus X (PVX) or potato virus Y (PVY). Third, the aphids aggregated preferentially on screening over leaflet models treated with volatiles collected from PLRV-infected plants as compared with those collected from uninfected plants. The specific cues eliciting the aphid responses were not determined, but differences between headspace volatiles of infected and uninfected plants suggest possible ones. PMID- 11886637 TI - Manipulation of offspring sex ratio by second-mated female house wrens. AB - In 1973, Trivers and Willard proposed that offspring sex ratio should be associated with the quality of parental care likely to be provided to the offspring. We tested this hypothesis by comparing fledgling sex ratios in nests of first- and second-mated female house wrens (Troglodytes aedon). In our Wyoming population, second-mated females typically receive little or no male parental assistance and fledge fewer and lower-quality young compared with first-mated females. Assuming that being of lower quality has stronger negative effects on the future reproductive success of males than that of females in this polygynous population, we predicted that fledgling sex ratios in the nests of second-mated females would be female-biased compared with the fledgling sex ratios of first mated females. Additionally, we asked whether any sex bias at fledging could have resulted from male-biased nestling mortality caused by sex-biased parental provisioning. As predicted, mean fledgling sex ratios in nests of second-mated females were more female-biased than fledgling sex ratios in nests of first-mated females. However, we found no evidence of either sex-biased nestling mortality or sex-biased parental provisioning. These findings suggest that females are responding to their status as second-mated females and to the associated low quality parental care that their young are likely to receive by producing female biased clutches rather than manipulating the offspring sex ratio through sex biased nestling mortality. PMID- 11886638 TI - Efficiency of gamete usage in nature: sperm storage, fertilization and polyspermy. AB - Gamete production for both males and females can be energetically expensive such that selection should maximize fertilization opportunities while minimizing fertilization costs. In laboratory studies of Drosophila reproduction, however, the failure of eggs to yield adult progeny can be quite high, suggesting that female control over gamete utilization is surprisingly inefficient. We examined gamete utilization in D. pseudoobscura from nature and compared our observations to those for laboratory populations. In natural populations 100% of oviposited eggs effectively produce adult progeny, and fertilization is exclusively monospermic, indicating that in nature, D. pseudoobscura females maintain a very strict control over their reproduction such that gamete usage is extremely efficient. The potential reasons for the inefficient gamete utilization in the laboratory, as well as the potential impact on laboratory studies of sperm competition, sexual conflict, and the evolution of reproductive barriers are discussed. Furthermore, in this sperm-heteromorphic species, our observations show definitively that in nature, as well as in the laboratory, only the long sperm morph participates in fertilization. PMID- 11886639 TI - A possible non-sexual origin of mate preference: are male guppies mimicking fruit? AB - In most animals, the origins of mating preferences are not clear. The "sensory bias" hypothesis proposes that biases in female sensory or neural systems are important in triggering sexual selection and in determining which male traits will become elaborated into sexual ornaments. Subsequently, other mechanisms can evolve for discriminating between high- and low-quality mates. Female guppies (Poecilia reticulata) generally show a preference for males with larger, more chromatic orange spots. It has been proposed that this preference originated because it enabled females to obtain high-quality mates. We present evidence for an alternative hypothesis, that the origin of the preference is a pleiotropic effect of a sensory bias for the colour orange, which might have arisen in the context of food detection. In field and laboratory experiments, adult guppies of both sexes were more responsive to orange-coloured objects than to objects of other colours, even outside a mating context. Across populations, variation in attraction to orange objects explained 94% of the inter-population variation in female mate preference for orange coloration on males. This is one of the first studies to show both an association between a potential trigger of a mate-choice preference and a sexually selected trait, and also that an innate attraction to a coloured inanimate object explains almost all of the observed variation in female mate choice. These results support the "sensory-bias" hypothesis for the evolution of mating preferences. PMID- 11886640 TI - In vivo regulation of the mandibular organ in the edible crab, Cancer pagurus. AB - Considerable evidence indicates that methyl farnesoate (MF) production by the crustacean mandibular organs is negatively regulated by neuropeptides from the sinus gland (SG) in the eyestalk. In the crab Cancer pagurus, two neuropeptides (MO-IH-1 and -2) have been isolated from the SG that inhibit MF synthesis by mandibular organs of female crabs in vitro. To test their activity in vivo, we treated eyestalk-ablated male crabs with SG extracts (SGEs) or MO-IH-1 and -2. SGEs reduced haemolymph levels of MF by 60-80%, while MO-IH-1 and -2 had little effect. Protease treatment of SGEs destroyed the in vivo activity, suggesting that the extract contains an additional peptide responsible for the in vivo activity. When separated by reversed-phase high performance liquid chromatography (HPLC), the in vivo activity eluted in fractions prior to MO-IH-1 and -2. When mandibular organs were removed from animals previously treated in vivo with these active fractions, they had reduced levels of MF synthesis and activity of farnesoic acid O-methyl transferase compared with mandibular organs from animals treated with saline. Together, these results indicate that the regulation of the crustacean mandibular organ is complex and may involve several SG compounds. Some of these compounds (i.e., MO-IH-1 and -2) act directly on the tissue while others affect the mandibular organ indirectly. PMID- 11886641 TI - Patterns in fish radiation are compatible with Pleistocene desiccation of Lake Victoria and 14,600 year history for its cichlid species flock. AB - Geophysical data are currently being interpreted as evidence for a late Pleistocene desiccation of Lake Victoria and its refilling 14,600 years ago. This implies that between 500 and 1000 endemic cichlid fish species must have evolved in 14,600 years, the fastest large-scale species radiation known. A recent review concludes that biological evidence clearly rejects the postulated Pleistocene desiccation of the lake: a 14,600 year history would imply exceptionally high speciation rates across a range of unrelated fish taxa. To test this suggestion, I calculated speciation rates for all 41 phylogenetic lineages of fish in the lake. Except for one cichlid lineage, accepting a 14 600 year history does not require any speciation rates that fall outside the range observed in fishes in other young lakes around the world. The exceptional taxon is a lineage of haplochromine cichlids that is also known for its rapid speciation elsewhere. Moreover, since it is unknown how many founding species it has, it is not certain that its speciation rates are really outside the range observed in fishes in other young lakes. Fish speciation rates are generally faster in younger than in older lakes, and those in Lake Victoria, by far the largest of the young lakes of the world, are no exception. From the speciation rates and from biogeographical observations that Lake Victoria endemics, which lack close relatives within the lake basin, have such relatives in adjacent drainage systems that may have had Holocene connections to Lake Victoria, I conclude that the composition of the fish assemblage does not provide biological evidence against Pleistocene desiccation. It supports a hypothesis of recent colonization from outside the lake basin rather than survival of a diverse assemblage within the basin. PMID- 11886642 TI - The X chromosome is a hot spot for sexually antagonistic fitness variation. AB - Sexually antagonistic alleles are selected discordantly between the sexes. Experimental evidence indicates that sexually antagonistic fitness variation is abundant in the genome of Drosophila melanogaster. Theory predicts that the X chromosome will be enriched with this type of variation. To test this prediction in D. melanogaster, we sampled, and cytogenetically cloned, 20 X chromosomes and compared their fitness variation to genome-wide levels. At the juvenile stage, in which gender roles are most similar, the X chromosome made no detectable contribution to genome-wide fitness variation. At the adult stage, in which gender roles diverge, the X chromosome was estimated to harbour 45% of the genome wide fitness variation and 97% of the genome-wide sexually antagonistic variation. This genomic structure has important implications for the process of sexual selection because X-linked sexually antagonistic variation contributes to negative intersexual heritability for fitness, i.e. high-fitness males (females) produce, on average, low-fitness daughters (sons). PMID- 11886643 TI - Trawling damage to Northeast Atlantic ancient coral reefs. AB - This contribution documents widespread trawling damage to cold-water coral reefs at 840-1300 m depth along the West Ireland continental shelf break and at 200 m off West Norway. These reefs are spectacular but poorly known. By-catches from commercial trawls for deep-water fish off West Ireland included large pieces (up to 1 m(2)) of coral that had been broken from reefs and a diverse array of coral associated benthos. Five azooxanthellate scleractinarian corals were identified in these by-catches, viz. Desmophyllum cristagalli, Enallopsammia rostrata, Lophelia pertusa, Madrepora oculata and Solenosmilia variabilis. Dating of carbonate skeletons using (14)C accelerator mass spectrometry showed that the trawled coral matrix was at least 4550 years old. Surveys by remotely operated vehicles in Norway showed extensive fishing damage to L. pertusa reefs. The urgent need for deep-water coral conservation measures is discussed in a Northeast Atlantic context. PMID- 11886644 TI - High temporal frequency synchrony is insufficient for perceptual grouping. AB - We used textures of randomly moving grating patches to assess the role of fine grain temporal synchrony in texture segregation. In the target area, patches reversed direction simultaneously. In the surround, patches changed direction at random times. Thus, phase changes in the target area were precisely synchronous, whereas those in the surround were not. In agreement with work carried out by Lee and Blake, we found that the target area was frequently visible, and that observers could discriminate its shape (horizontal versus vertical) at frame rates of 100 Hz in brief exposures (200 ms). Further experiments suggested that the length of unidirectional motion sequences in the target area, rather than synchrony, determined its visibility. To eliminate completely contrast and motion cues, we made all the background elements identical to the target elements, but with a random starting phase. Despite the presence of synchrony in the target area but not the background, the target was generally very hard to see. Targets that remained visible contained low temporal frequency modulations of direction. We conclude that the human observer can detect synchrony, but only at modest temporal frequencies once motion and contrast artefacts have been eliminated. PMID- 11886645 TI - Prehistoric bird extinctions and human hunting. AB - Holocene fossils document the extinction of hundreds of bird species on Pacific islands during prehistoric human occupation. Human hunting is implicated in these extinctions, but the impact of hunting is difficult to disentangle from the effects of other changes induced by humans, including habitat destruction and the introduction of other mammalian predators. Here, we use data from bones collected at a natural sand dune site and associated archaeological middens in New Zealand to show that, having controlled for differences in body mass and family membership (and hence for variation in life-history traits related to population growth rate), birds that were more intensively hunted by prehistoric humans had a higher probability of extinction. This result cannot be attributed to preservation biases and provides clear evidence that selective hunting contributed significantly to prehistoric bird extinctions at this site. PMID- 11886646 TI - Oystercatchers use colour preference to achieve longer-term optimality. AB - The optimal diet model entails that foragers look beyond the individual prey encounter, to at least the level of intake rate across a bout of foraging, but optimization over a longer time remains controversial. In this paper, we show how oystercatchers increase their intake over the longer term using mussel colour as a cue. Wintering oystercatchers Haematopus ostralegus feed extensively on mussels Mytilus edulis in the estuaries of southern Britain. They show a marked preference for brown-shelled mussels over the commoner black-shelled morph, and we show that this enables them to maximize their rate of energy gain over a longer period than a single foraging bout. The brown and black mussels did not differ in ventral thickness and energy content, which are the main criteria for mussel selection and most important for short-term optimization. The brown mussels contained significantly less moisture, so by selecting them, oystercatchers could pack more mussel flesh into their limited oesophageal storage capacity. This enables them to increase their overall consumption during a feeding bout and increases their long-run energy gain rate, to an extent that is large enough to be significant for survival, especially during the short exposure of the mussel beds in winter. PMID- 11886647 TI - Parapoxvirus causes a deleterious disease in red squirrels associated with UK population declines. AB - The disease implications of novel pathogens need to be considered when investigating the ecological impact of species translocations on native fauna. Traditional explanations based on competition or predation may often not be the whole story. Evidence suggests that an emerging infectious disease, caused by a parapoxvirus, may be a significant component of the impact that the introduced grey squirrel has had on UK red squirrel populations. Here we validate the potential role of parapoxvirus by proving that the virus is highly pathogenic in the red squirrel while having no detectable effect on grey squirrel health. PMID- 11886648 TI - A view of early vertebrate evolution inferred from the phylogeny of polystome parasites (Monogenea: Polystomatidae). AB - The Polystomatidae is the only family within the Monogenea to parasitize sarcopterygians such as the Australian lungfish Neoceratodus poisteri and freshwater tetrapods (lissamphibians and chelonians). We present a phylogeny based on partial 18S rDNA sequences of 26 species of Polystomatidae and three taxon from the infrasubclass Oligonchoinea (= Polyopisthocotylea) obtained from the gills of teleost fishes. The basal position of the polystome from lungfish within the Polystomatidae suggests that the family arose during the evolutionary transition between actinopterygians and sarcopterygians, ca. 425 million years (Myr) ago. The monophyly of the polystomatid lineages from chelonian and lissamphibian hosts, in addition to estimates of the divergence times, indicate that polystomatids from turtles radiated ca. 191 Myr ago, following a switch from an aquatic amniote presumed to be extinct to turtles, which diversified in the Upper Triassic. Within polystomatids from lissamphibians, we observe a polytomy of four lineages, namely caudatan, neobatrachian, pelobatid and pipid polystomatid lineages, which occurred ca. 246 Myr ago according to molecular divergence-time estimates. This suggests that the first polystomatids of amphibians originated during the evolution and diversification of lissamphibian orders and suborders ca. 250 Myr ago. Finally, we report a vicariance event between two major groups of neobatrachian polystomes, which is probably linked to the separation of South America from Africa ca. 100 Myr ago. PMID- 11886649 TI - The design of scaffolds for use in tissue engineering. Part II. Rapid prototyping techniques. AB - Tissue engineering (TE) is an important emerging area in biomedical engineering for creating biological alternatives for harvested tissues, implants, and prostheses. In TE, a highly porous artificial extracellular matrix or scaffold is required to accommodate mammalian cells and guide their growth and tissue regeneration in three-dimension (3D). However, existing 3D scaffolds for TE proved less than ideal for actual applications because they lack mechanical strength, interconnected channels, and controlled porosity or pores distribution. In this paper, the authors review the application and advancement of rapid prototyping (RP) techniques in the design and creation of synthetic scaffolds for use in TE. We also review the advantages and benefits, and limitations and shortcomings of current RP techniques as well as the future direction of RP development in TE scaffold fabrication. PMID- 11886650 TI - Evaluation of Na+ active transport and morphological changes for bioartificial renal tubule cell device using Madin-Darby canine kidney cells. AB - The function of current hemodialysis as an artificial kidney is insufficient because of the lack of reabsorptive function. In this study, we intend to develop a bioartificial renal tubule cell device (RTD) using tubular epithelial cells and artificial membranes and to evaluate the reabsorptive function of the confluent layers. Madin-Darby canine kidney (MDCK) cells were cultured on a nucleopore polycarbonate membrane for up to 4 weeks after confluence to examine the influence of the culture period on their ability to transport Na+ actively using Na+/K+ATPase (NKA). The results were (1) active Na+ transport of the cells averaged 24.8 mM/m(2) x 24 h during the initial 2 weeks after confluence and then decreased to about 4.2 mM/m(2) x 24 h during the next 2 weeks; (2) NKA localized on the basal-lateral sides of the cells during the initial 2 weeks, whereas it also localized on the apical side of the cells during the next 2 weeks; (3) long term culture resulted in an increased number of upheaving cell mass, increased fatty droplets in the cells, and necrosis; and (4) scanning electron microscopy showed fewer microvilli 3-four weeks after confluence. It is concluded that the culture period is critical for developing RTD using cultured renal tubular epithelial cells. PMID- 11886651 TI - Oxygen and inulin transport measurements in a planar tissue-engineered bioartificial organ. AB - In vivo oxygen and inulin transport rates were measured in a planar tissue engineered bioartificial organ implanted in a rat. A compartmental model was used to describe the transport of oxygen and inulin between the cell chamber, across the immunoisolation membrane, and within the neovascularized region adjacent to the immunoisolation membrane. A nonlinear regression analysis of the plasma inulin levels and the oxygen transport rate into the device provided information on the degree of vascularization in the region adjacent to the bioartificial organ. Key parameters that were obtained from the analysis of the in vivo transport data included the average capillary blood oxygen partial pressure, the Krogh tissue cylinder radius, the extracellular volume fraction, and the capillary blood residence time. These four parameters are important indicators for assessing the degree of vascularization in the tissue adjacent to the immunoisolation membrane in the bioartificial organ. The oxygen and inulin transport technique reported here is a useful tool for describing the in vivo transport characteristics of a bioartificial organ and for assessment of the vascularization within tissue engineered structures. PMID- 11886652 TI - Proliferating cells versus differentiated cells in tissue engineering. AB - The efficiency of cell or tissue cultures is usually judged by how quickly confluence is reached within a Petri dish or on a scaffold. Growth factors and fetal bovine serum are employed to drive cultured cells from one mitosis to the next as quickly as possible. The tissue specific interphase is extremely short under these conditions, so that the degree of differentiation desired in tissue engineering cannot be achieved. To reach the goal of functional differentiation in vitro mitosis and interphase must be separated experimentally and tailored to the specific requirements of the cell-type used. This could be achieved by a three step concept for tissue-engineering in vitro as we present here. The expansion phase is followed by a phase in which tissue differentiation is initiated. The final phase serves to express and maintain histotypical differentiation of the generated tissue. PMID- 11886653 TI - Salt fusion: an approach to improve pore interconnectivity within tissue engineering scaffolds. AB - Macroporous scaffolds composed of biodegradable polymers have found extensive use as three-dimensional substrates either for in vitro cell seeding followed by transplantation, or as conductive substrates for direct implantation in vivo. Methods abound for creation of macroporous scaffolds for tissue engineering, and common methods typically employ a solid porogen within a three-dimensional polymer matrix to create a well-defined pore size, pore structure, and total scaffold porosity. This study describes an approach to impart improved pore interconnectivity to polymer scaffolds for tissue engineering by partially fusing the solid porogen together prior to creation of a continuous polymer matrix. Three dimensional, porous scaffolds of the copolymer 85:15 poly(lactide-co glycolide) were fabricated via either a solvent casting/particulate leaching process, or a gas foaming/particulate leaching process. Prior to creation of a continuous polymer matrix the NaCl crystals, which serve as the solid porogen, are partially fused via treatment in 95% humidity. Scanning electron micrographs clearly display fused salt crystals and an enhancement in pore interconnectivity in the salt fused scaffolds prepared via both solvent casting and gas foaming, and the extent of pore interconnectivity is enhanced with longer treatment times. Fusion of salt crystal for 24 h increased the radius of curvature of salt crystals, and led to a twofold increase in the compressive modulus of solvent cast scaffolds (total porosity of 97 +/- 1%). Fusion of NaCl crystals prior to gas foaming resulted in a decrease in scaffold compressive modulus from 277 +/- 60k Pa to 187 +/- 30k Pa (total porosity of 94 +/- 1%). The resulting highly interconnected scaffolds have implications for facilitated cell migration, abundant cell-cell interaction, and potentially improved neural and vascular growth within tissue engineering scaffolds. PMID- 11886654 TI - The Th2-restricted immune response to xenogeneic small intestinal submucosa does not influence systemic protective immunity to viral and bacterial pathogens. AB - Implantation of mice with xenogeneic extracellular matrix (ECM) not only results in tissue remodeling but also elicits a strong Th2 immune response. It is possible that the Th2 cytokines induced by ECM act systemically and result in immune suppression to unrelated antigens. In this case, the recipient would be predisposed to immune dysfunction and have increased susceptibility to various pathogens. The purpose of this study was to determine if ECM implantation does, in fact, influence the immune response to other antigens. Four models were examined to determine the effects of ECM implantation on systemic immunity. In the first model, mice were subcutaneously implanted with porcine small intestinal submucosa (SIS) and immunized with a T-dependent subunit vaccine against influenza virus. The antibody response and protection against lethal infection were then measured. The second model consisted of similar experiments performed using a T-independent polysaccharide vaccine against S. pneumoniae. In the third model, mice were implanted and the cell-mediated response to dinitrofluorobenzene (DNFB) challenge was determined. The fourth model involved examining the influence of SIS implantation on rejection of xenogeneic skin grafts. We found that antibody levels of mice vaccinated against influenza virus or S. pneumoniae were not affected by SIS implantation and these mice did not exhibit increased or decreased susceptibility to either infectious agent. Similarly, mice implanted with ECM showed no cell-mediated immune dysfunction upon challenge with DNFB or xenogeneic skin grafts. The results of this study demonstrate that the Th2 restricted response induced by xenogeneic ECM implantation does not cause generalized immune suppression. Therefore, SIS implantation does not increase susceptibility to viral or bacterial pathogenic agents. PMID- 11886655 TI - Antimicrobial activity associated with extracellular matrices. AB - Materials derived from extracellular matrices (ECMs) are being evaluated as scaffolds for surgical reconstruction of damaged or missing tissues. It is important to understand the susceptibility of these biological materials to bacterial infections. ECMs derived from porcine small intestinal submucosa (SIS) and urinary bladder submucosa (UBS) were found to possess antimicrobial activity. ECM extracts, obtained by digesting these acellular matrices in acetic acid, demonstrated antibacterial activity against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Antimicrobial activity was determined using a minimal inhibitory concentration assay. Bacteriostatic activity was detected at protein concentrations of ECM extracts equivalent to 0.77-1.60 mg/mL. ECM extracts were found to inhibit bacterial growth for up to at least 13 h. The resulting extracts consisted of water-soluble peptides and proteins with molecular weights ranging from <4 to >100 kDa and lower molecular weight compounds, as determined by size exclusion liquid chromatography. PMID- 11886656 TI - Differential effects of growth factors on tissue-engineered cartilage. AB - The effects of four regulatory factors on tissue-engineered cartilage were examined with specific focus on the ability to increase construct growth rate and concentrations of glycosaminoglycans (GAG) and collagen, the major extracellular matrix (ECM) components. Bovine calf articular chondrocytes were seeded onto biodegradable polyglycolic acid (PGA) scaffolds and cultured in medium with or without supplemental insulin-like growth factor (IGF-I), interleukin-4 (IL-4), transforming growth factor-beta1 (TGF-beta1) or platelet-derived growth factor (PDGF). IGF-I, IL-4, and TGF-beta1 increased construct wet weights by 1.5-2.9 fold over 4 weeks of culture and increased amounts of cartilaginous ECM components. IGF-I (10-300 ng/mL) maintained wet weight fractions of GAG in constructs seeded at high cell density and increased by up to fivefold GAG fractions in constructs seeded at lower cell density. TGF-beta1 (30 ng/mL) increased wet weight fractions of total collagen by up to 1.4-fold while maintaining a high fraction of type II collagen (79 plus minus 11% of the total collagen). IL-4 (1-100 ng/mL) minimized the thickness of the GAG-depleted region at the construct surfaces. PDGF (1-100 ng/mL) decreased construct growth rate and ECM fractions. Different regulatory factors thus elicit significantly different chondrogenic responses and can be used to selectively control the growth rate and improve the composition of engineered cartilage. PMID- 11886657 TI - Tissue-engineered human auricular cartilage demonstrates euploidy by flow cytometry. AB - Transforming growth factor-beta (TGF-beta) and basic fibroblast growth factor (bFGF) are known to stimulate the rate of chondrocyte proliferation. The theoretical risk of malignant transformation associated with growth factor stimulation of chondrocytes should be addressed; aneuploidy has been found to occur in human cartilaginous tumors. In this study, chondrocytes were obtained from six human auricles and cultured in vitro for 6 weeks in the presence or absence of TGF-beta and bFGF. Cells were analyzed for DNA at 3-, 4-, 5-, and 6 week intervals by flow cytometry (FACScan), which demonstrated no evidence of aneuploidy. A persistent increase in S-phase was noted in cells cultured only with TGF-beta. Cells were implanted in athymic mice, and after 8 weeks of implantation, the cartilage constructs formed were examined histologically. The tissue-engineered cartilage cultured originally in bFGF most resembled normal, native cartilage. Specimens cultured in TGF-beta produced suboptimal cartilage morphology. Flow cytometry shows no evidence of aneuploidy, with chondrocytes maintaining their normal diploid state. Further studies incorporating additional methods of analysis need to be done. PMID- 11886658 TI - Formation of vascularized meniscal tissue by combining gene therapy with tissue engineering. AB - Ingrowth of host blood vessels into engineered tissues has potential benefits for successful transplantation of engineered tissues as well as healing of surrounding host tissues. In particular, the use of a vascularized bioengineered tissue could be beneficial for treating injuries to the meniscus, a structure in the knee where the lack of a vascular supply is associated with an inadequate healing response. In this study, gene transfer using an adenovirus vector encoding the hepatocyte growth factor gene (AdHGF) was used to induce blood vessel formation in tissue-engineered meniscus. Bovine meniscal cells were treated with AdHGF, a vector encoding a marker gene E. coli beta-galactosidase (Adbetagal), or no virus. Cells were seeded onto poly-glycolic acid felt scaffolds and then transplanted into the subcutaneous pouch of athymic nude mice for 8 weeks. Expression of the marker gene and HGF was detectable for several weeks after gene transfer. Ink injection studies showed that AdHGF-treated meniscal cells formed tissue which contained fourfold more blood vessels at 2 weeks (p < 0.02) and 2.5-fold more blood vessels at 8 weeks (p < 0.001) posttransplantation than controls. This study demonstrates the feasibility of using adenovirus-mediated gene transfer to engineer a blood supply in the bioengineered meniscal tissue. PMID- 11886659 TI - Genetic engineering of meniscal allografts. AB - Allograft meniscal transplantation represents one of the few available treatment options after menisectomy. Despite acceptable early results, a considerable controversy exists with regard to poor graft regeneration, shrinkage and biomechanical failure of transplanted menisci. Transfer of specific growth factor genes may improve the regeneration process of meniscal allografts. The aim of this study was to investigate the feasibility of gene transfer in meniscal allografts in rabbits. Four different viral vectors encoding marker genes, including lacZ, luciferase, and green fluorescence protein were used to investigate viral transduction in 50 lapine menisci for 4 weeks in vitro. Subsequently, 16 unilateral meniscus replacements were performed with ex vivo retrovirally transduced meniscal allografts, and the expression of the lacZ gene was examined histologically at 2, 4, 6, and 8 weeks after transplantation. Gene expression in the superficial cell layers of the menisci can be detected for up to 4 weeks in vitro, but the level of gene transfer declined over time. The transduction with retrovirus showed better persistence and deep penetration of the menisci with infected cells. In vivo, declining numbers of beta-galactosidase positive cells were also detected in retrovirally transduced allografts up to 8 weeks. Consistently, transduced cells were found at the menisco-synovial junction of the transplants and in deeper layers of the menisci. There was no evidence of cellular immune response in the transduced transplants. This investigation showed a prospective for growth factor delivery in auto- and allografts. In further experiments, vectors expressing therapeutic proteins such as growth factors will be investigated to assess their potential to improve remodeling and healing of meniscal allografts. PMID- 11886660 TI - Effect of glycosaminoglycan production on hardness of cultured cartilage fabricated by the collagen-gel embedding method. AB - To assess applicability of the tactile sensor in hardness measurement of cultured cartilage and to clarify the relationship between hardness and tissue structure of cultured cartilage fabricated by the collagen-gel embedding method, we studied the effect of glycosaminoglycans on hardness of such cultured cartilage using a tactile sensor and electron probe x-ray microanalyzer (EPMA). Hardness measured by the tactile sensor, that is, change in frequency of naturally oscillating piezoelectric elements caused by contact with a testing material, increased with the number of days of culture or seeding cell density. Analysis of the sulfur component in EPMA results mainly reflected glycosaminoglycans produced by chondrocytes. Sulfur mapping indicated that tissue of the cultured cartilage differed between its surface and the inside; layers rich in glycosaminoglycans and cells had formed in the surface. Changes in frequency showed close correlation with the amount of glycosaminoglycans in the surface and the inside (r = 0.98 and 0.85, respectively) of cultured cartilage measured by EPMA. Thus, the tactile sensor is capable of measuring hardness of cultured cartilage, reflecting the change in tissue structure between the surface and the inside of the cartilage. PMID- 11886661 TI - In vitro engineered cartilage constructs produced by press-coating biodegradable polymer with human mesenchymal stem cells. AB - Cartilage constructs were fabricated by press-coating D,D-L,L-polylactic acid polymer blocks of 1 x 0.5 x 0.5 cm onto high-density cell pellets of 1.5 x 10(6) human mesenchymal stem cells (mhMSCs) isolated from the femoral head of patients undergoing total hip arthroplasty. Following attachment of the cell pellets to the polymer surfaces, chondrogenesis was induced by culturing the constructs for 3 weeks in a serum-free, chemically defined, chondrogenic differentiation medium supplemented with transforming growth factor beta-1 (TGF-beta1). Histochemical analysis showed that the press-coated pellets formed cell layers composed of morphologically distinct, chondrocyte-like cells, surrounded by a fibrous, sulfated proteoglycan-rich extracellular matrix. Immunohistochemical analysis detected collagen type II and cartilage proteoglycan link protein within the extracellular matrix. Expression of the cartilage-specific marker genes collagen types II, IX, X, and XI, and aggrecan was detected by RT-PCR. Scanning electron microscopy revealed organized and spatially distinct zones of cells within the cell-polymer constructs, with the superficial layer resembling compact hyaline cartilage. The fabrication method of press-coating biodegradable polymers with mhMSCs allows the in vitro production of cartilage constructs without harvesting chondrocytes from intact articular cartilage surfaces. These constructs may be applicable as prototypes for the reconstruction of articular cartilage defects in humans. PMID- 11886662 TI - Bone marrow tissue engineering. AB - The creation of mixed hematopoietic chimerism has become an important clinical strategy for tolerance induction for cellular and organ transplantation, and for the treatment of numerous hematopoietic diseases. Clinical success has been limited however, by host immune response and by competition from host hematopoiesis. Recent data suggests that limited donor stem cell engraftment after minimally myeloablative hematopoietic stem cell (HSC) transplantation may in part be due to MHC associated microenvironmental mismatch resulting in a competitive disadvantage for donor HSC. A strategy to overcome this barrier to stable mixed hematopoietic chimerism would involve concurrent transplantation of a donor bone marrow microenvironment. To test this possibility, we set out to develop a method to tissue engineer a bone marrow microenvironment. One to two murine femurs were mechanically crushed to a fine suspension and were combined in vitro with various delivery vehicles. These constructs were transplanted into syngeneic animals in locations that are known to support transplantation of other tissues. Although bone formation was observed with several conditions, bone marrow formation was noted only within the small bowel mesentery when type I collagen was used as the delivery vehicle. No bone marrow formed when the vehicle was changed to polyglycolic acid or type IV collagen. We have demonstrated that the small bowel mesentery can support bone marrow formation under specific in vivo conditions. Future work will focus on strategies for transplantation of an engineered donor bone marrow environment to facilitate creation of allogeneic mixed hematopoietic chimerism. PMID- 11886664 TI - Where we are in telemedicine/telehealth, and where we go from here. PMID- 11886663 TI - Endogenous adipocyte precursor cells for regenerative soft-tissue engineering. AB - Subcutaneous injection of reconstituted basement membrane (Matrigel) in combination with basic fibroblast growth factor induces de novo adipogenesis in which endogenous precursor cells invade the artificially formed Matrigel space, proliferate and differentiate to form adipose tissue. Since this adipogenesis offers us a novel approach for soft-tissue reconstruction without transplanting preadipocytes, the early process was examined by optical and electron microscopy. Formation of multiple layers of fibroblast-like cells at the surface of Matrigel implant was the first response of connective tissue. The cells within four to five layers proximal to Matrigel implant acquired a thick cytoplasm and an enlarged nucleus, and they invaded Matrigel space together with endothelial cells which caused neovascularization. Phagocytotic incorporation and digestion of Matrigel components by well-developed lysosomes appeared to be a stimulus of fibroblast-like cells to mature depending on proximity to Matrigel. The fibroblast-like cells often contacted to the outer surface of capillary over a large area and rapidly accumulated lipid droplets. Electron microscopy of the developing adipocytes showed a well-organized smooth endoplasmic reticulum and mitochondria. This investigation thus revealed the characteristics of adipocyte precursor cells, which can be recruited for regenerative engineering of soft tissues. PMID- 11886665 TI - Issues and problems before us. PMID- 11886666 TI - Improvement in asthma symptoms and quality of life in pediatric patients through specialty care delivered via telemedicine. AB - Asthma is the most common chronic disease affecting children. Studies have demonstrated improvements in asthma control when care is delivered by specialists compared with generalists. We postulated that specialist care delivered by telemedicine would result in similar improvements in control of symptoms and quality of life as compared with face-to-face encounters with specialists. Seventeen patients with persistent asthma, who were cared for by pediatricians in a rural school-based health clinic, were treated over a 6-month period in an asthma specialty program. Patients had face-to-face encounters at week zero, and then telemedicine follow-up visits at weeks 4, 12, and 24. Patients maintained a symptom diary and reliever medication use log. Spirometry and patient and caregiver quality-of-life questionnaires were completed at each visit. Mean number of symptom free days increased 83% from 2.35 days at week 0 to 4.31 days at week 24 (p < 0.05). There was a 44% reduction in mean symptom scores, from 2.32 at week 0 to 1.31 at week 24 (p < 0.001). Nine patients reported having 7 symptom-free days or 7 days of symptom scores of zero in the preceding seven days at week 24 compared with one patient at week 0 (p < 0.002). FEV(1) increased by > or = 12% in seven patients during the study period. Significant improvements in quality of life were reported by patients at week 4 (p < 0.02) and week 24 (p < 0.01), and by caregivers at week 24 (p < 0.002). Specialty asthma care delivered via telemedicine resulted in improvements in asthma symptom control and quality of life similar to improvements reported in face-to-face encounters provided by specialists. PMID- 11886667 TI - Limitations of patient satisfaction studies in telehealthcare: a systematic review of the literature. AB - The objective of this study is to provide a systematic review of studies on patient satisfaction with telemedicine. The review included empirical studies that investigated patient satisfaction with that telemedicine service. The search strategy involved matching at least one of 11 'telemedicine' terms with one of 5 'satisfaction' terms. The following databases were searched: Telemedicine Information Exchange (TIE) database, MEDLINE, Science Citation Index (SCI), Social Science Citation Index (SSCI), Psycinfo, and Citation Index of Nursing and Allied Health (CINAHL). A highly structured instrument was used for data extraction. The review included 93 studies. Telepsychiatry represents the largest portion of these studies (25%), followed by multispecialty care (14%), nursing (11%), and dermatology (8%). Real-time videoconferencing was used in 88% of these studies. Only 19 (20%) included an independent control group, including 9 (10%) randomized control trial (RCT) studies. One third of studies were based on samples of less than 20 patients, and only 21% had samples of over 100 patients. Aspects of patient satisfaction most commonly assessed were: professional-patient interaction, the patient's feeling about the consultation, and technical aspects of the consultation. Only 33% of the studies included a measure of preference between telemedicine and face-to-face consultation. Almost half the studies measured only 1 or 2 dimensions of satisfaction. Reported levels of satisfaction with telemedicine are consistently greater than 80%, and frequently reported at 100%. Progression of telemedicine services from "trial" status to routine health service must be supported by improved research into patients' satisfaction with telemedicine. Further investigation of factors that influence patient acceptance of telemedicine is indicated. PMID- 11886668 TI - The usefulness of telemedicine for the detection of infection/inflammation at the point of care. AB - The objective of this study is to examine the possibility of using Telemedicine to diagnose the presence of the inflammatory response and to assess its intensity at the point of care. One drop of citrated peripheral venous blood from 15 patients with infection/inflammation and 15 controls were used to prepare the slides. Unstained pictures were analyzed using a microscope, video camera and image analyzer (INFLAMETTM, Biovision, Tel Aviv, Israel). The jpg-compressed images were transferred via telephone to a physician in a remote location. A significant correlation was noted between the white blood cell count and the number of leukocytes per square mm by image analysis (r = 0.67 p < 0.0001 n = 30), between the degree of leukocyte adhesiveness/aggregation and the concentration of C-reactive protein (r = 0.42 p = 0.02 n = 29) and between the degree of erythrocyte aggregation and either fibrinogen concentrations (r = 0.73 p < 0.0001) or erythrocyte sedimentation (r = 0.83, p < 0.0001). No problems occurred during file transmission and there were no transfer errors. Physicians can successfully estimate the presence of an inflammatory response and its intensity using a simple slide test, image analysis, and Telemedicine technology. PMID- 11886669 TI - A virtual instrument for acquisition and analysis of the phonocardiogram and its internet-based application. AB - The objective of this study is to develop a phonocardiogram (PCG) acquisition and analysis instrument using virtual instrumentation technology and investigate its Internet-based application. The PCG instrument was developed using a Pentium 200 computer, a data acquisition board, and a two-channel custom designed bio-signal preamplifier. LabVIEW was used to create the instrument's front panels. Spectral and joint time-frequency analyses were implemented into the instrument. This instrument can be used to display the PCG and to analyze the individual heart sound and murmur for the detection of heart valve diseases. Using a test-bed, the PCG data acquisition and analysis were performed remotely over the Internet. Through the main PCG panel, an operator can control the acquisition and analysis of PCG signals. In the remote test, real-time transmission of the PCG signal over the Internet was possible. Remote operators were able to view smoothly scrolling PCG waveforms and could control all the acquisition parameters and perform spectral and time-frequency analyses on the acquired heart sound. This study demonstrated that a LabVIEW-based medical virtual instrument provides a low-cost and flexible solution for data acquisition and analysis of PCG. It also showed that the current Internet supports the transmission of real-time PCG signals. Compared with other telemedicine systems, this application transfers not only the medical data, but also the virtual instrument and its signal processing capability through the Internet. PMID- 11886670 TI - Remote percutaneous renal access using a new automated telesurgical robotic system. AB - Previous clinical application of remote telesurgery has been the use of a novel system of video teleconferencing equipment along with remote control of a laparoscopic camera at distances over 11,000 miles. Recently, a robotic system has been developed to assist with percutaneous renal surgery. This robot has been incorporated into the telesurgical system to allow remote needle placement into the renal collecting system under radiological guidance. The main component of the telesurgical system is a low degree of freedom robot called "PAKY" (percutaneous access of the kidney). It is custom designed for fluoroscopic guided percutaneous needle insertion into the renal collecting system. The robot is a six-degrees of freedom device. However, when the skin entry site is fixed and held in position, only two degrees of freedom are required to orient the needle in the correct plane for accurate insertion. Remote control of the robot was accomplished over a plain old telephone system (POTS) line. On June 17, 1998, the first remote telerobotic percutaneous renal access procedure was performed between the Johns Hopkins Hospital, Baltimore, Maryland, and Tor Vergata University, Rome, Italy. This new telesurgical robot was successful in term of obtaining percutaneous access within 20 min, with two attempts to obtain entry into the collecting system. This robot represents the first system for performing remote telesurgical interventions in the kidney and demonstrates the feasibility and safety of assisting accurate and rapid needle access to the kidney during percutaneous procedures. PMID- 11886671 TI - [DEBONEL (Dermacentor-borne-necrosis-erythemalymphadenopathy). A new tick-borne disease?]. PMID- 11886673 TI - [Effectiveness of contact isolation in the control of multiresistant bacteria in an intensive care service]. AB - AIM: To describe the frequency, characteristics and progression of critically ill patients admitted to the ICU, for whom isolation is indicated due to detection of multiresistant pathogenic bacteria, and to study the effectiveness of precautionary measures to avoid dissemination of these microorganisms. PATIENTS AND METHODS: Prospective, observational, cohort study performed by a specially created working group of four nurses and an ICU specialist. The study included 55 patients in whom contact isolation was indicated (isolation rate, 15.2 per 100 patients), collected over a 16-month period. RESULTS: The multiresistant bacteria responsible for isolation of the patients were: Pseudomonas aeruginosa (17 cases), Staphylococcus aureus (17 cases), Stenotrophomonas maltophilia (15 cases), Acinetobacter baumannii (4 cases) and extended-spectrum beta-lactamase (ESBL)- producing Enterobacteria (2 cases). Vancomycin-resistant Enterococcus spp. was not identified in any case. The mean duration of ICU isolation was 17.6 6 5.1 days (range 1-75). Multiresistant bacteria were classified as intra-ICU nosocomial in 39 cases (70.9%), extra-ICU nosocomial in 10 cases (18.2%) and community-acquired in 6 (10.9%). During the study period, no epidemic outbreak due to any of the controlled bacteria was detected. The multiresistant bacteria presented in the form of colonization in 41 cases (74.5%). The reasons for discontinuing isolation were death of the patient in 18 cases, transferal to a hospital ward (discharge from the ICU) in 19 cases, and eradication of the bacteria in 18 cases. Of the 55 patients with multiresistant bacteria, 35 (63.6%) died during hospitalization, and 23 of these (41.8%) during their stay in the ICU. CONCLUSIONS: The implementation of a working team for early detection of multiresistant pathogenic bacteria resulted in application of contact isolation in 15.2% of patients admitted. Surveillance to fulfill isolation precautions in a medical-surgical ICU achieved an absence of epidemic outbreaks due to these bacteria during the study period. PMID- 11886672 TI - [Oral and cervicofacial actinomycosis. Presentation of five cases]. AB - BACKGROUND: Actinomycosis is a suppurative and granulomatous chronic infectious disease caused by Actinomyces sp. and most commonly affecting the cervicofacial area. AIM: To study the clinical characteristics of patients with actinomycosis, with regard to clinical history, presentation, method of diagnosis, treatment and follow up. PATIENTS AND METHODS: A retrospective review was performed on all cases of microbiologically or histologically proven oral or cervicofacial actinomycosis. RESULTS: Five patients were studied, 2 men and 3 women, 25-69 years old. Four patients had a history of surgical procedures and/or dental manipulations. Three patients showed the classic presentation of a lump and fistulization, and two patients presented intra-oral lesions. Four patients were diagnosed by cultures positive to A. israelii on microbiologic study and the remaining patient by cytologic detection of a sulfur granule. The first patient received the classic initial regimen of iv penicillin and 3 were treated with third-generation cephalosporins, continuing with oral amoxicillin during 12 months. Patient no.2 required a second surgical procedure. Patient no. 5, who had an exclusively oral process, received a short course of amoxicillin. There were no relapses during follow-up. CONCLUSION: Actinomycosis is an uncommon disease. Establishment of the definite diagnosis requires a high index of suspicion and good clinical-microbiological collaboration. The classic course of iv penicillin and oral amoxicillin during 6-12 months is effective. For the acute phase treatment, iv penicillin can be replaced by third-generation cephalosporins. PMID- 11886674 TI - [Seroprevalence of hepatitis C virus in the general population]. AB - AIM: To determine the prevalence of hepatitis C virus (HCV) infection and to study associated risk factors in the general population of the province of Zamora. PATIENTS AND METHODS: For this transversal, observational, descriptive, population study, we randomly selected 1973 individuals over 14 years old and grouped them according to age and sex. The study included completion of a questionnaire containing socio-demographic information and data on risk factors. Serum samples were collected for analysis of antibodies to HCV by means of a third-generation ELISA, and were confirmed by an immunoblot technique. Viral load and genotype analyses were carried out in positive cases by polymerase chain reaction technique. RESULTS: Among the total, 600 complete studies (questionnaires and serum samples) 657 questionnaires and 675 samples were obtained. Eleven serum samples tested positive for HCV and among these, 5 were confirmed by immunoblot. The positive samples corresponded to 3 men and 2 women; 4 lived in urban settings and all 5 had one or more known risk factors for acquiring the infection. CONCLUSIONS: The prevalence of HCV infection in our geographical area was 0.74% (95% CI, 0.27-1.82). The seropositive individuals detected in this population screening study had a history of parenteral risk activities and most lived in urban areas. PMID- 11886675 TI - [Characteristics of tuberculosis in a general hospital during the period 1993 1998. Analysis of resistance and HIV coinfection]. AB - AIM: The epidemiologic and clinical characteristics, presence of HIV coinfection, and sensitivity to tuberculostatic drugs were analyzed in a series of tuberculosis patients attended in our center. PATIENTS AND METHODS: Retrospective study of tuberculosis cases attended in a third- level hospital from 1993 to 1998. RESULTS: During the study period, 268 cases of tuberculosis were diagnosed in our center. A progressive decrease in the incidence of this disease has occurred since 1995. Among the jailed population, we also found a decrease in cases of tuberculosis and there were no cases of resistance. In the total population, only 8 isolates (3.27%) showed resistance to some of the antituberculosis drugs studied (isoniazid, rifampicin, ethambutol, streptomycin). A tendency toward a decrease in resistance was also observed starting from 1995, with no new cases detected in the last two years. There was a 1.3% rate of primary resistance to isoniazid. Multiresistance was detected in only 4 patients, two of whom died. The rate of HIV coinfection was 38.8%. In 39% of cases the form of presentation was exclusively pulmonary and in 25% it was disseminated. CONCLUSIONS: There was a 50% decrease in tuberculosis cases during the period studied. The rate of HIV coinfection was 38.8%, one of the highest in the literature, indicating that HIV serology should be included in the protocol for studying tuberculosis in our setting. Given the low rate of resistance detected, we recommend a three-drug regimen for antituberculosis treatment. PMID- 11886676 TI - [Infections in joint prostheses: epidemiology and clinical presentation. A prospective study 1992-1999]. AB - BACKGROUND: The prosthetic infection is a serious complication due to diagnostic problems. AIM: To determinate epidemiological characteristics, and clinical patterns of infections associated to prosthetic materials to improve the diagnosis and management. PATIENTS AND METHODS: From december 1992 to december 1999, 110 patients with prosthetic infections were prospectively evaluated. Diagnosis was made according to standard microbiological, clinical and radiological criteria. RESULTS: The incidence was 5.1% (110/1,400) prosthetic materials insert in the period of study. The average age was 59.6 years (range 18 79), and the majority of patients 63 (57.2%) were female. Forty-two (38%) suffered a total knee replacement, 29 (26%) a total hip replacement, 1 (1%) shoulder replacement and 38 (34%) autogenous bone gratting. In 29 patients (26.3%) a previous chronic disease had been diagnosed (diabetes, neoplasis, rheumatoid arthritis). Previous use of antibiotics was detected in 58 patients (51%), being ciprofloxacin the most frequently used. An etiological diagnosis was reached in 66 patients (60%), isolated grampositive in 58.2%, gramnegative in 32.8%, fundamentally by Staphylococcus sp. and P. aeruginosa respectively. In 9% anaerobe were isolated. There were early infections in 67 cases, delayed in 25, and late in 18. All the patients had local pain and flogotiv signs as initial findings, whereas 46 (41.8%) developed osteocutaneous fistula and only 5 (4.5%) presented temperature. CONCLUSIONS: Prosthetic infection is a frequent complication after articular replacement, and grampositive cocci predominate as ethilogical agents. Sistemic clinical manifestations are uncommon. PMID- 11886677 TI - [Variability in the prescription of antibiotics]. PMID- 11886678 TI - [Bacteriemia in a girl with rhabdomyosarcoma]. PMID- 11886679 TI - [Frequent, acute episodes of normocytic anemia in a patient with AIDS]. PMID- 11886680 TI - [Lumbar tumor in a patient with AIDS]. PMID- 11886681 TI - [Contribution to diagnosis of microbiology results from samples obtained in the Emergency Room of a Children's Hospital]. PMID- 11886682 TI - [Isolation of Mycobacterium lentiflavum in a case of suspected lung cancer]. PMID- 11886683 TI - [Candida tropicalis meningitis associated with external ventricular drainage in an adult female patient]. PMID- 11886684 TI - [Isolated laryngeal leishmaniasis and bone marrow culture]. PMID- 11886685 TI - [Mesenteric thrombosis associated with cytomegalovirus infection in an immunocompetent patient]. PMID- 11886686 TI - [Hepatitis C virus and type-2 diabetes mellitus. Is there a connection?]. PMID- 11886687 TI - [Mixed bacterial infections of the oral cavity]. PMID- 11886688 TI - Global benefit-risk assessment of antidepressants: venlafaxine XR and fluoxetine. AB - BACKGROUND: Quantifying efficacy and safety differences between drugs is difficult because rigorous statistical methods to assess benefit and risk simultaneously are lacking. METHODS: Global benefit-risk (GBR) analysis of clinical trial data was used retrospectively to compare venlafaxine extended release (XR) and fluoxetine. Of 301 outpatients with moderate to severe depression given venlafaxine XR 75-225 mg/day (n=100), fluoxetine 20-60 mg/day (n=103), or placebo (n=98) for up to 8 weeks, 295 qualified for analysis. Primary efficacy variables were Hamilton Rating Scale for Depression (HAM-D) remission (final on-therapy score risk category were 2.1 (1.1-4.0) and 2.2 (1.1 4.3) for venlafaxine XR vs. fluoxetine and placebo, respectively. For CGI response, relative gains of venlafaxine XR were 1.39 (P<0.01) and 1.45 (P<0.01) vs. fluoxetine and placebo; benefit exceeded risk in 66, 53, and 52% of patients given venlafaxine XR, fluoxetine, and placebo (P=0.041 vs. venlafaxine XR), respectively. CONCLUSIONS: GBR analysis can be applied to a wide array of efficacy and safety data to form statistical tests of clinically meaningful treatment comparisons. In this comparison, the GBR assessments on response and remission significantly favored venlafaxine XR. PMID- 11886689 TI - Molecular characterisation of antidepressant effects in the mouse brain using gene expression profiling. AB - Antidepressants are widely used for the treatment of psychiatric disorders, including depression and anxiety. Although they are efficient drugs, there are several unsolved questions regarding their clinical pharmacology. Furthermore, the molecular mechanisms of action of antidepressants are still poorly understood and the molecular targets and pathways remain to be identified. To address these issues, we performed a gene expression analysis in mice treated with two commonly used antidepressants with differing pharmacology (paroxetine or mirtazapine) for 1, 7 or 28 days. We quantified the effects of these treatments on gene expression in the mouse brain with cDNA-microarrays containing 3624 expressed sequence tags (ESTs) representing murine genes expressed in the brain. We found that both drugs led to downregulation of four common genes. In addition, although it was possible to identify common targets for the two drugs, the expression profiles of the drugs differed in a fundamental manner, and the longer the treatment duration, the greater the difference in the profiles. These findings suggest that antidepressants with different pharmacologies can share molecular targets even though the primary pathways at which they act are different. PMID- 11886690 TI - Relation between responses to repetitive transcranial magnetic stimulation and partial sleep deprivation in major depression. AB - Repetitive transcranial magnetic stimulation (rTMS) has been found to ameliorate symptoms in major depression. However, its mechanism of action has to be further elucidated and the relationship between responses to rTMS and other antidepressant interventions except electroconvulsive therapy has not been investigated to date. Here we studied in an open trial whether the response to partial sleep deprivation may predict the clinical outcome of rTMS treatment. Thirty-three drug-free patients suffering from a major depressive episode underwent a partial sleep deprivation at least 5 days prior to rTMS and subsequently received 10 sessions of 10 Hz rTMS of the left prefrontal cortex. After rTMS a significant overall improvement of 32% on the Hamilton Rating Scale for Depression was observed. Forty-two percent of patients showed an antidepressant response after rTMS. Amelioration of depression after partial sleep deprivation was inversely correlated with improvement after rTMS. There was no clinically applicable predictive value of the response to partial sleep deprivation for the outcome after rTMS. Apparently, different subgroups of depressed patients respond to both interventions. Further studies are needed to characterize the response to rTMS by means of clinical and biological parameters. PMID- 11886691 TI - Biological nature of depressive symptoms in borderline personality disorder: endocrine comparison to recurrent brief and major depression. AB - Borderline personality disorder (BPD) often shows depressive symptoms and their biological nature albeit extensively discussed remains controversial. The knowledge of this nature seems essential as it could imply key therapeutic strategies. We have found BPD and major depression (MD) not to share biological abnormalities. We have proposed BPD to frequently display an affective syndrome distinct from the nonborderline MD both in terms of quality and duration of symptoms and of biological substrate. A substantial number of BPD patients can be diagnosed as having clinical Recurrent Brief Depression (RBD) which has been proposed to overlap with BPD. RBD has been found to share perturbed biological substrate with MD but we have previously not found this abnormal substrate in BPD. Our aim was to study the possibility that BPD patients with depressive symptoms and even clinically diagnosed with RBD have a biological substrate distinct from RBD without BPD and from MD, and therefore an specific affective syndrome. We compared 20 BPD in-patients without co-existing MD to 20 sex- and age-matched non-BPD recurrent brief depressives and to 20 sex- and age-matched non-BPD major depressives on the thyrotropin-releasing hormone stimulation test (TRH-ST) and the dexamethasone suppression test (DST). Twelve BPD patients were diagnosed as having also RBD. BPD had less TRH-ST blunting than MD. TRH-ST did not differentiate BPD from RBD. RBD and MD patients shared equivalent TRH-ST values but BPD patients with clinically diagnosed RBD did not. BPD and RBD showed less perturbed DST than MD. DST did not differentiate BPD from RBD. BPD and RBD share most of the endocrinological normal substrate already described in BPD but RBD also share abnormalities with MD. Whereas we can conceptualize RBD as being an endocrinologically perturbed depressive syndrome, this may not be the case for the possible specific affective syndrome of BPD even if it can be for now diagnosed as being RBD. PMID- 11886693 TI - Intent-to-treat analysis for clinical trials: use of data collected after termination of treatment protocol. AB - Following patients for a period of time after termination of treatment protocols is a common practice in clinical trials of drug treatments. After termination of the protocol treatment, patients are usually provided with medical care as deemed appropriate, e.g. different doses of the same drug, augmentation with other drugs, or different drugs. Data collected on patients when they are not in the treatment protocol are termed "off-treatment" data to be differentiated from "on treatment" data that are collected while patients are in the treatment protocol. The purpose of the present paper is to describe some recent statistical methodological advances in the use of "off-treatment" data for intent-to-treat (IT) analysis using mixture models [Biometrics 52 (1996) 1002]. Two-piece spline models conditional on the protocol treatment dropout time are developed first. The two pieces of the spline model represent the on-treatment and off-treatment segments of data and are joined at the dropout time. The weighted average of the two-piece conditional models across realizations of the dropout times provides the mixture model for the pragmatic IT analysis. The weights are the estimates of the probabilities of the dropout times. The mixture model is amenable to an explanatory analysis which assumes that the patients remain on their assigned treatments. The model allows the parameters of the splines to depend on the dropout time. Statistical inference is based on bootstrap sampling procedures. We have illustrated this methodology using data from a drug trial comparing nortriptyline and paroxetine in the treatment of major depression in older patients. PMID- 11886692 TI - Mirtazapine, yohimbine or olanzapine augmentation therapy for serotonin reuptake associated female sexual dysfunction: a randomized, placebo controlled trial. AB - BACKGROUND: Many agents have been proposed as potential treatments for SSRI associated sexual dysfunction, but few placebo-controlled trials have been reported. METHOD: After a 1-month baseline evaluation, pre-menopausal women with moderate to severe sexual dysfunction associated with the institution of fluoxetine therapy were randomized to augmentation therapy with placebo (N=39), mirtazapine (N=36), yohimbine (N=35) or olanzapine (N=38) for a 6-week period. Outcomes were measured using a daily diary, a biweekly self-report assessment, and a computer assisted structured interview. RESULTS: At baseline, orgasm was most severely impaired. After 6 weeks, there was statistically significant improvement on most measures for the overall group of patients, however there were few differences between treatment groups. Isolated treatment differences were observed for the patient self-report of overall sexual function (olanzapine superior to placebo) and the structured interview sexual satisfaction item (mirtazapine inferior to placebo). CONCLUSION: No drug assessed was consistently associated with differences from placebo. The results of the study do not support uncontrolled reports of efficacy for these agents in premenopausal women. PMID- 11886694 TI - The relation of "acute and transient psychotic disorder" (ICD-10 F23) to bipolar schizoaffective disorder. AB - The aim of this work is to investigate differences between acute and transient psychotic disorders (ATPD; F23 of ICD-10) and bipolar schizoaffective disorders (BSAD). In a controlled prospective and longitudinal study, we compared all inpatients with ATPD treated at Halle university hospital during a 5-year period with matched controls with BSAD. Sociobiographical data were collected using a semi-structured interview. Follow-up investigations were performed at a mean of 2.2-3.3 years after the index episode or 8.2-16.1 years after the first episode by means of standardized instruments. ATPD differs significantly from BSAD on various relevant levels, such as gender (more female), age at onset (older), development of the full symptomatology (more rapid), duration of the symptomatology (shorter), acuteness of onset (more acute), preceding stressful life-events (more frequent) and long-term prognosis (better). It is concluded that ATPD and BSAD are different nosological entities. PMID- 11886695 TI - Urinary free cortisol and childhood trauma in cocaine dependent adults. AB - OBJECTIVE: To examine whether childhood trauma may have a relationship to hypothalamic-pituitary-adrenal (HPA) axis function as an adult. METHODS: Forty six withdrawn cocaine dependent patients participated in 24-h urine collections for determination of urinary-free cortisol (UFC) and completed the Childhood Trauma Questionnaire (CTQ). RESULTS: Patients with a mean UFC output below the median had significantly higher CTQ scores for childhood sexual abuse than patients with UFC outputs above the median. Multiple regression analysis showed that both childhood emotional neglect and sexual abuse were independently associated with UFC outputs. CONCLUSION: These cross sectional data, in a sample of middle-aged cocaine dependent patients, suggest the possibility that childhood trauma may have an effect on HPA axis function and thus predispose to psychiatric disorders. Further studies are needed in different samples. PMID- 11886696 TI - The effect of guided imagery and amitriptyline on daily fibromyalgia pain: a prospective, randomized, controlled trial. AB - OBJECTIVE: The effectiveness of an attention distracting and an attention focusing guided imagery as well as the effect of amitriptyline on fibromyalgic pain was studied prospectively. METHODS: Fifty-five women with previously diagnosed fibromyalgia were monitored for daily pain (VAS) in a randomized, controlled clinical trial. One group received relaxation training and guided instruction in "pleasant imagery" (PI) in order to distract from the pain experience (n=17). Another group received relaxation training and attention imagery upon the "active workings of the internal pain control systems", "attention imagery" (AI) (n=21). The control group (CG) received treatment as usual (n=17). Patients were also randomly assigned to 50-mg amitriptyline/day or placebo. Some psychological and socio-demographic variables were also measured initially. The slopes of diary pain ratings over a 4-week period were used as the outcome measures. RESULTS: We found significant differences of the pain-slopes between the three psychological conditions (P=0.0001). The pleasant imagery (P<0.005), but not the attention imagery group's slope, declined significantly when compared with the control group (P>0.05). There was neither a difference between the amitriptyline and placebo slopes (main effects, P=0.98) nor a significant amitriptyline x psychological interaction (P=0.76). CONCLUSION: Pleasant imagery (PI) was an effective intervention in reducing fibromyalgic pain during the 28-day study period. Amitriptyline had no significant advantage over placebo during the study period. PMID- 11886697 TI - Nocturnal secretion of TSH and ACTH in male patients with depression and healthy controls. AB - Profound alterations of the hypothalamic-pituitary-thyroid (HPT) and the hypothalamic-pituitary-adrenal (HPA) systems at the hypophyseal level have been described in affective disorder. To precisely characterize the basal alterations of both axes during sleep, we simultaneously investigated sleep EEG and the secretion of thyrotropin, ACTH and cortisol in nine drug-free male patients with depression in comparison to 10 healthy age and sex matched controls. In depressed patients the nearly diametrical nocturnal secretion of thyrotropin and ACTH was disturbed by significantly blunted thyrotropin values (TSH AUC 51.96+/-5.68 vs. 87.23+/-13.63, P<0.05) and elevated ACTH values (ACTH AUC 1804+/-161 vs. 1538+/ 130, P<0.05) compared to controls. Moreover, cross correlation analysis revealed a highly negative association of 0 lag between thyrotropin and ACTH and between thyrotropin and cortisol in the control sample, indicating a physiological nocturnal negative correlation of HPT and HPA system. In the patients sample these associations were weak and reached not statistical significance. Therefore, as a descriptive tool, the ratio TSH/ACTH revealed a significant group difference between controls and patients in the first half of the night (TSH/ACTH AUC 6.50+/ 0.42 vs. 3.35+/-0.31, P<0.05). Sleep-EEG analysis showed a shortened REM latency, a decrease of stage 2 and an increase of awake time in the patients. Our data support the hypothesis that both hypophyseal hormones reflect a common dysregulation of both systems in depression probably due to impaired action of TRH-related corticotropin-release-inhibiting-factor (CRIF). The ratio TSH/ACTH might be a tool to characterize alterations of both the HPT and HPA axis in depression during the first half of the night. PMID- 11886698 TI - Generalized linear least squares algorithms for modeling glucose metabolism in the human brain with corrections for vascular effects. AB - The generalized linear least squares (GLLS) algorithm has been found useful in image-wide parameter estimation for the generation of parametric images with positron emission tomography (PET) as it is computationally efficient and statistically reliable. However, the original algorithm was designed for parameter estimation with non-uniformly sampled instantaneous measurements. When dynamic PET data are sampled with the optimal image sampling schedule (OISS) to reduce memory and storage space, only a few temporal image frames are recorded. As a result, the direct application of GLLS is no longer appropriate. In this paper, we extend the GLLS algorithm to a five parameter model for the study of human brain metabolism, which accounts for the effect of cerebral blood volume (CBV), using OISS sampled data, with as few as five temporal samples. The formulation for this new GLLS algorithm is developed, and its computational efficiency and statistical reliability are investigated and validated using computer simulations and clinical PET [18F]-2-fluoro-2-deoxy-D-glucose (FDG) data. PMID- 11886699 TI - How to avoid misinterpretation of heart rate variability power spectra? AB - Spectral analysis of R-R Interval time series is increasingly used to determine periodic components of heart rate variability (HRV). Particular diagnostic relevance is assigned to a low-frequency (LF) component, associated with blood pressure regulation, and a high-frequency (HF) component, also referred to as respiratory sinus arrhythmia (RSA) in the HRV power spectra. Frequency ranges for parametrisation of power spectra have been defined for either component in numerous publications.Results obtained from examinations with standardised psychic load in which ECG and respiratory signal are continuously recorded and adequately processed have shown that the true individual frequency range of the HF component can be reliably determined only by means of characteristics of respiration (respiratory rate (RR), range and median value of RR, tidal depth). Respiratory rhythms are interindividually extremely differentiated and of individual-specific nature. In many cases LF and HF components may be totally superimposed on each other and, consequently, cannot be diagnostically evaluated. PMID- 11886700 TI - Automatic recognition of cell layers in corneal confocal microscopy images. AB - A confocal microscope can produce gray-scale images of the different layers of the cornea. We have addressed the problem of classifying these images, i.e. recognizing the layer displayed, using the shape of the cells contained, which is uniquely related to each specific layer. A first method was designed, based first on the binarization of the image and then on the description of the cell shape by means of Hu variables (central moments). An artificial neural network was used to classify each image according to the values assumed by these variables. A Matlab prototype of the classification system was developed, considering images of three corneal layers (Bowman membrane, stroma, endothelium) in normal subjects. The system was tested on 46 images, and good results were obtained. To avoid the critical step of binarization, an alternative cell shape description was investigated, based on Zernike moments, and a new network was developed and trained. The results achieved were better than those obtained with the previous technique, and also, no binarization was necessary. PMID- 11886701 TI - Coupling patterns between spontaneous rhythms and respiration in cardiovascular variability signals. AB - We performed a quantitative study of coupling patterns between respiration and spontaneous rhythms of heart rate and blood pressure variability signals by using the Recurrence Quantification Analysis (RQA). We applied RQA to both simulated and experimental data obtained in control breathing at three different frequencies (0.25, 0.20, and 0.13 Hz) from ten normal subjects. RQA succeeded in quantifying different degrees of non-linear coupling associated to several interference patterns. We found higher degrees of non-linear coupling when the respiratory frequency was close to the spontaneous Low Frequency (LF) rhythm (0.13 Hz), or almost twice the LF frequency (0.2 Hz), whereas weaker coupling was observed when the respiratory frequency was 0.25 Hz. Clinical applications of our approach should focus on new experimental protocols, featuring the stimulation of one of the two branches of the autonomic nervous system (ANS) or aimed at the analysis of pathologies linked to the ANS. PMID- 11886702 TI - Complexity of biomedical data models in cardiology: the Intranet-based AF registry. AB - Atrial fibrillation (AF) is the most common cardiac arrhythmia and is associated with major complications. Ongoing research is focused on new pacing devices for alternative treatment of this disease. The objective of an AF registry is to store prospectively all relevant data covering clinical information, quality of life and device parameters and by this means provide a platform for long-term follow-up. For statistical analysis, categorical and numerical items are required, thus a high-granular data structure must be defined and implemented in the clinical setting. Facing the limits of formalization, we developed an XML based documentation scheme consisting of 619 items in eight tables and implemented it with state-of-the-art Intranet technology. At present detailed information on 88 patients is recorded. The pacing device generates per patient and follow-up visit a file consisting of approximately 400-500 parameters provided on a floppy disk, which are transferred by means of a specific interface into the database. Success factors for integration of a complex research database into the routine workflow of a busy university hospital are interfaces between data sources to enable non-redundant data entry, intensive fine tuning by iterative software engineering and benefit for the clinical users in form of clinical reports and patient-specific summaries. Data quality must be assured by plausibility checks. To get an overview of this complex dataset we developed a dedicated visualization tool. Due to the high number of items a large patient collective must be recruited for statistical evaluation. Interinstitutional cooperation is required for a consensus on common minimal documentation schemes to enable pooling of data. PMID- 11886703 TI - Unified algorithm for real-time detection of drug interaction and drug allergy. AB - This algorithm aims at unifying and generalizing the algorithm for detecting all types of documented drug interactions such as drug-drug interactions, drug disease interactions, drug-patient interactions (drug allergy) from patient profile information and drug-laboratory test interactions in real-time prescribing system. Ideally, the system should conform to the following criteria: (1) data independence; (2) software interconnectability; (3) knowledge expandability; (4) flexibility; and (5) computation resource efficiency. We propose a robust Structured Query Language (SQL) algorithm to detect drug interactions and drug allergy according to such criteria. We believe that this is the first public domain algorithm in SQL that could be easily implemented into most open-system prescribing software which support SQL language. The algorithm comprises two major stages: 'expand' and 'extract'. The former expands all information in the prescription with their synonyms, groups, or components. The latter extracts the documented interactions by inner-joining knowledge-base with two independent copies of the expanded prescription list simultaneously. Simulation study for speed performance indicates that this algorithm is well behaved, for the speed of computation does not grow faster than the growth in prescription size. PMID- 11886704 TI - Which coding system for therapeutic information in evidence-based medicine. AB - The coding of information in the computer representation of clinical trials is essential both for the rationalisation of the activities involved in the production of therapeutic information for evidence-based decision support and for the integration of the messages produced by these activities with clinical information and electronic patient record systems. There is no standard coding system available, however, so building on existing evaluations, we performed a simple semi-quantitative evaluation of ICD-10, CDAM, MEDDRA, MESH, READ, SNOMED and UMLS to provide objective criteria for the choice of a coding system. Inclusion and exclusion criteria for four clinical trials recorded in TriSum constituted the corpus of evaluation texts. Criteria included coding coverage, size, integration and language coverage. The results of the comparison lead us to choose SNOMED as the most appropriate coding system for our needs. The absence of a European Medical Language System project is observed, as is the need for combinatorial as opposed to enumerative systems. PMID- 11886705 TI - The ups and downs of the impact factor: the case of Archives of Medical Research. PMID- 11886706 TI - The oral side of Sjogren syndrome. Diagnosis and treatment. A review. AB - Sjogren syndrome (SS) is an inflammatory disease of the exocrine glands. Although not always present, signs and symptoms of dry eyes and xerostomia are characteristic features of SS. Oral dryness is one of the most important data of patients with SS. Several sets of criteria have been published; however, there is no definitive agreement concerning which is the most useful. In addition to its various clinical manifestations, lack of understanding of the causes of SS delays prompt diagnosis. Histologically, the salivary gland shows a characteristic lymphocytic infiltrate, which is implicated in the destruction of gland cells. Saliva performs an important role in maintaining and protecting oral health. Deficient quality and quantity of saliva have a detrimental consequence for dental and oral health. In some patients, appropriate information regarding dry mouth care is not offered because most professionals either neglect or ignore adequate attention to oral health. Therefore, lack of treatment is frequent. Medical and dental studies that focus on the oral aspects of diagnosis, consequences, and treatment of SS are commented on. Diagnostic methods used for the oral component are also reviewed. The role of the oral tests developed to diagnose SS is assessed, especially tests used by the majority of criteria. Impairment of salivary secretion increases the risk of developing oral diseases; the therapeutic modalities designed to ameliorate these damages by increasing salivary output or by substitution of saliva are reviewed. We discuss published prevention techniques to diminish dental, periodontal, and soft tissue infections. PMID- 11886707 TI - In vitro proliferation, expansion, and differentiation of a CD34+ cell-enriched hematopoietic cell population from human umbilical cord blood in response to recombinant cytokines. AB - BACKGROUND: The conditions and mechanisms that control the in vitro growth of hematopoietic stem/progenitor cells (contained within the population of CD34+ cells) are still not completely understood. METHODS: By using an immunomagnetic system, we have enriched for umbilical cord blood (UCB)-derived CD34+ cells (55% of total cells recovered vs. 0.8% of total cells prior to the enrichment procedure) and analyzed their in vitro growth (proliferation, expansion, and differentiation) in a liquid culture system in the absence or presence of different recombinant cytokine combinations. RESULTS: When the selected cells were cultured in the absence of recombinant cytokines, no proliferation or expansion was observed. In the presence of steel factor (SF) and interleukin-6 (IL-6), total cell number was increased nearly fourfold; however, no progenitor cell expansion took place. When cultures were supplemented with SF and IL-6 together with IL-3 and erythropoietin (EPO), a rapid proliferation of the CD34+ enriched cell population was observed with a selective stimulation of erythropoiesis. However, this stimulation was only transient, suggesting that there was a rapid exhaustion of erythroid progenitor cells within the first 10 days. Significantly higher levels of proliferation and expansion of progenitor cells were observed in the presence of SF, IL-6, GM-CSF, and G-CSF with preferential stimulation of myelopoiesis. Interestingly, such stimulation of myelopoiesis was sustained for the entire culture period (>30 days). The highest levels of proliferation and expansion were observed in the presence of all six cytokines. Under these conditions, erythropoiesis was also sustained only transiently (10 days), whereas myelopoiesis was sustained for >30 days. CONCLUSIONS: This study indicates that significant proliferation and expansion of hematopoietic progenitors can be achieved in vitro when culturing a cell population in which CD34+ cells comprise only >50% of the total cells. Our results also suggest that myeloid progenitors (those responding to GM-CSF and G CSF) possess higher expansion potentials in vitro than their erythroid counterparts. The methods described here for the enrichment and culture of CD34+ cells may be relevant in the development of protocols for the ex vivo proliferation and expansion of hematopoietic progenitors for transplantation. PMID- 11886708 TI - Superoxide-superoxide oxidoreductase activity of the captopril-copper complex. AB - BACKGROUND: The purpose of this study was to determine whether the interaction of captopril, an angiotensin-converting enzyme inhibitor, with copper could modify the superoxide dismutase activity of this metal. Results may help to explain the interaction of captopril with reactive oxygen species in the stunned myocardium where substantial mobilization of copper and iron in the coronary flow following ischemia has been reported. METHODS: An assay that generates superoxide anion radicals without the intervention of metal ions was utilized. In addition, direct EPR analysis was applied to assess the redox state of copper during reactions. RESULTS: Captopril-copper complex inhibited the superoxide-mediated reduction of nitroblue tetrazolium. In addition, captopril-copper complex was able to suppress formazan production by potassium superoxide. Direct EPR analysis showed that copper was reduced to the cuprous state by captopril and remained in this state in the course of the reaction. Captopril was also stable during the dismutation reaction. CONCLUSIONS: We conclude that cuprous-captopril complex is a catalytic species with properties different from those of Cu(2+) alone. A model in which sulfur acts as electron acceptor/donor in place of the metal is proposed and a mechanism of action for this complex is discussed. PMID- 11886709 TI - Human lymphocyte antigen DR7 protects against proliferative retinopathy with type II diabetes mellitus. AB - BACKGROUND: This study was undertaken in order to analyze the genetic incidence of human lymphocyte antigen diabetic retinopathy (HLA-DR) and its influence in proliferative diabetic retinopathy (PDR). METHODS: We designed a case-control study in which 127 mestizo Mexican patients with DM II and diabetic retinopathy were studied. DNA was extracted and HLA-DR regions were amplified using PCR. Alleles were determined by DNA hybridization. Diagnosis was assessed clinically and by fluorescein angiography. Incidence of HLA-DR alleles in patients was compared with an ethnically matched control group of healthy subjects (n = 98). Statistical significance was established with non-parametric tests. RESULTS: Patients with diabetic retinopathy showed less frequency of HLA-D11 compared with the control group (p = 0.043). NPDR patients with 10 or more years of DM II showed an increase of HLA-DR7 (p = 0.01). CONCLUSIONS: Our results suggest that the presence of HLA-DR7 protects against the development of proliferative disease in the diabetic Mexican population. PMID- 11886710 TI - DNA damage and repair in lymphoblastoid cell lines from normal donors and fragile X syndrome patients. AB - BACKGROUND: Because lymphocytes from fragile X patients have been reported as hypersensitive to bleomycin-induced chromatid breaks and because the number of trinucleotide repeats in families with fragile X syndrome has a propensity to expand, we have investigated the possibility that fragile X cells may be hypersensitive to DNA damage and have a lower capacity for DNA repair. METHODS: Lymphocytes from normal and fragile X syndrome donors were immortalized by Epstein-Barr virus transformation. Characteristics of fragile X syndrome including the folate-sensitive fragile site on chromosome Xq27.3, length of CGG repeat expansion, and FMRP expression in Epstein-Barr virus-transformed lymphoblastoid cell lines were analyzed by standard cytogenetic methods, Southern blot, and Western blot, respectively. Analysis of DNA damage and repair induced by hydrogen peroxide, bleomycin, ethyl methanesulfonate, 4-nitroquinoline-N oxide, etoposide, and mitomycin C was carried out by single-cell gel electrophoresis assay (known as comet assay). RESULTS: Lymphoblastoid cell lines from fragile X donors had a folate-sensitive fragile site on chromosome Xq27.3, no or low FMRP expression, and expansion of the CGG repeat. Results of comet assay showed that fragile X cells were not more sensitive to mutagen-induced DNA strand breaks and did not have lower DNA repair capacity in comparison with normal cells. Furthermore, one fragile X cell line showed hyposensitivity to DNA strand breaks induced by hydrogen peroxide, bleomycin, and ethyl methansulfonate. CONCLUSIONS: The results of this study do not support the notion that CGG trinucleotide expansion in fragile X syndrome is caused by permanent deficiency in DNA repair. PMID- 11886711 TI - Allogeneic peripheral blood stem cell transplantation. Safety, hematologic recovery, kinetics, and complications. AB - BACKGROUND: Hematopoietic stem cell transplants are an accepted treatment for several malignant and nonmalignant hematologic diseases. Recently, the peripheral blood, after mobilization of stem cells with growth factors, has become the source of choice for hematopoietic stem cells. We report on a series of patients who received peripheral blood stem cell transplants at the Instituto Nacional de Cancerologia (INCAN) in Mexico City. METHODS: Between May 1995 and December 1999, 33 patients received peripheral blood stem cell transplants to treat hematologic diseases. Sixty percent of our patients had chronic myelocytic leukemia (CML). All had a matched related donor. Patients were conditioned with one of five different conditioning regimens and subsequently received one of two different graft-vs.-host disease prophylaxis regimens. Stratified Wilcoxon rank-sum, chi square, and Mann-Whitney tests were used to analyze the results. RESULTS: In our series, median time to achieve a total neutrophil count of 0.5 x 10(9)/L was 14 days and to achieve a platelet count of 20 x 10(9)/L, 15 days. Acute graft-vs. host disease occurred in seven patients. Chronic graft-vs.-host disease occurred in 69% of surviving patients. Survival for low-risk patients was 67% and for the high-risk group, 9%. CONCLUSIONS: Peripheral blood stem cells produce a faster hematopoietic recovery. The rate of acute graft-vs.-host disease is not increased using the peripheral blood as source of stem cells; however, chronic graft-vs. host disease continues to be a significant problem. Donors tolerated the procurement procedure without complications. PMID- 11886712 TI - Increased incidence of anxiety and depression during bone marrow transplantation. AB - BACKGROUND: Since the first organ transplantation in the 1950s, there have been reports that patients who underwent organ transplantation had a poor prognosis if they were depressed and/or anxious prior to transplantation. Our objective in this study was to determine the prevalence of anxiety and depression in the different stages of bone marrow transplantation (BMT). METHODS: Mood disorders (MD), anxiety disorders (AD), and adjustment disorders (ADD) were measured five times with DSM IV. The Beck Inventory of Depression (BID) and Hamilton Rating Scale of Anxiety (HRSA) were used to measure levels of depression and anxiety, respectively, at registration and at days -1, +21 +/- 7, +30 +/- 10, and 90 +/- 10 days from BMT. Analysis between diverse periods was made for allogeneic BMT (allo-BMT) with chi square test, while Fisher exact test was used for the autologous BMT (auto-BMT). RESULTS: We report on 26 patients, including 18 with allo-BMT, and eight with auto-BMT. The allo-BMT was associated with depression during post BMT period (chi(2) = 11.924; p = 0.01). Slight anxiety without statistical significance was found in all stages. There was a high prevalence of anxiety and adjustment disorder in the immediate posttransplantation stage. Anxiety and adjustment disorders were more frequently found in all posttransplantation stages, particularly in the immediate stage (chi(2) =11.104, p = 0.02). After 3 months, no survivor received a psychiatric diagnosis. We did not find any differences in MD. There were five deaths. CONCLUSIONS: The auto-BMT group did not show significant associations between different stages and psychiatric variables studied. One death occurred at 1 month. This patient had severe depression. We recommend that the depressive syndrome be intentionally researched during the different stages of BMT, specifically in the immediate transplantation stage. PMID- 11886713 TI - Enhanced sister-chromatid exchange rate in human lymphocytes exposed to 17beta estradiol in vitro. AB - BACKGROUND: Epidemiologic data and animal experiments strongly implicate that steroid hormones are involved in the process of malignant transformation due to their capability to stimulate mitotic division and/or elevate the level of mutations in susceptible cells. METHODS: The objectives of this investigation were to evaluate the effects of 17beta estradiol in sister-chromatid exchange (SCE) test on cultured human peripheral blood lymphocytes. The lowest concentration of 17beta estradiol used in this experiment (10(-10) M) was calculated as comparable with the physiologic blood level of 17beta estradiol in women. Three experimental concentrations corresponded to minimal (7 x 10(-8) M), average (3.5 x 10(-6) M), and maximal (7 x 10(-6) M) therapeutic doses in human medicine. In addition, the highest concentrations exceed maximal therapeutic dose 10-fold (7 x 10(-5) M) and 30-fold (2.1 x 10(-4) M), respectively. RESULTS: The obtained results indicate that estradiol significantly elevates SCE per cell frequency at all concentrations applied except at the lowest one. However, estradiol has not influenced mitotic activity of cultured human lymphocytes significantly. CONCLUSIONS: It can be concluded that 17beta estradiol expressed genotoxic effects and therefore might represent a human health risk. PMID- 11886714 TI - Bone mineral density loss in patients with urolithiasis: a follow-up study. AB - BACKGROUND: Recurrent calcium urolithiasis is often associated with disorders of calcium metabolism. The purpose of this investigation was to assess bone mineral content (BMC) and bone mineral density (BMD) over a period of 1 year in patients with urolithiasis and to determine the factors that could have influenced the changes in bone density during that period. METHODS: The patient group comprised 34 men aged 41.2 plus minus 7.9 years with recurrent urolithiasis. A wide spectrum of biochemical measurements was performed. Bone mineral density (g/cm(2)), bone mineral content (BMC), and bone area (BA) were measured twice during a period of 1 year at the lumbar spine (L2-L4), femoral neck, Ward triangle, and trochanter, using dual energy absorptiometry. Patient results were compared to those obtained from 30 healthy male controls of a comparable age group. RESULTS: Nine patients were hypercalciuric, while the majority of the remaining metabolic parameters were within the reference values. Bone mineral content and bone areas at all regions were lower in patients comparing to controls, but not significantly. The greatest annual reduction of BMD was noticed at Ward triangle (-5.70% in patients and -2.36% in controls), followed by femoral neck (-4.06% patients, -2.03% controls) and trochanter (-3.06% patients, -1.39% controls). There was no significant decrease of the BMD of the spine. Analyzing the influence of age, body mass index (BMI), metabolic parameters, and dietary calcium intake on the annual reduction of bone density, we found that age, hyperuricosuria, and calcium intake were significantly associated with bone loss in that time period. CONCLUSIONS: Bone mass reduction in patients with urolithiasis over a 1-year period did not differ significantly from that in controls and was principally related to age, hyperuricosuria, and calcium dietary restriction but not to increased calcium excretion. PMID- 11886715 TI - Successful early pyeloplasty in infants. AB - BACKGROUND: Early pyeloplasty for the treatment of congenital ureteropelvic junction obstruction to maximize nephron salvage is justified only if potential hazards of operating on small infants are avoided. METHODS: The records were analyzed of all infants who underwent pyeloplasty over a 5-year period. Open pyeloplasty was performed if collecting systems had deteriorated or were demonstrated to be obstructed; it was also performed for severe cases of hydronephrosis. Outcome of surgery in the younger infant (patients <2 months of age) was compared with the older infant group (patients >2 months of age). Preoperative evaluation in case of mild or moderate hydronephrosis was directed toward ruling out a non-obstructed collection system and included voiding cystourethrography, and serial ultrasonography and/or dual isotope diuretic renography. Postoperative assessment consisted of serial ultrasonography and/or nuclear imaging to confirm decompression and relief of obstruction. RESULTS: A total of 24 pyeloplasties were performed on 22 patients in the younger infant group (two bilateral) and 30 were performed on 27 infants in the older infant group (three bilateral). The only significant differences between the groups were as follows: patients in the younger infant group were likely to present in utero (75%, p = 2.69), whereas those in the older infant group were more likely to present with a urinary tract infection (48%, p = 4.12). During follow-up examination, 23 renal units in the younger infant group and 24 in the older infant group were judged to be stable or improved. Four kidneys were not salvaged after pyeloplasty, one in the younger infant group and three in the older infant group. CONCLUSIONS: Good results of pyeloplasties performed in the infants in this series support early correction of ureteropelvic junction obstruction in infants. PMID- 11886716 TI - Population distribution residing at different altitudes: implications for hypoxemia. AB - BACKGROUND: Hypoxemia can adversely affect health. The present study is aimed at estimating the prevalence of altitude-related low partial pressure of oxygen in arterial blood in Mexican population by means of census and geographic data. METHODS: Population, altitude, and characteristics of communities were obtained from the Mexican Institute of Statistics, Geography, and Informatics (INEGI). The population of each municipality (municipio) was assumed to have the same age distribution as that reported for the entire country. Partial pressure of oxygen in arterial blood (PaO(2)) was estimated from altitude and from a hypothetical alveolar-arterial oxygen gradient that increases with age. RESULTS: In Mexico, 3.95% of the population lives at altitudes 2,500 m above sea level. Census data for the year 1990 recount 3.6 million people distributed among 6,150 communities. Exposure to intermediate altitudes is considerable: one half of the Mexican population resides above 1,550 m, 32% above 2,000 m, 25% above 2,230 m, and 5% above 2,440 m. It was estimated that between 0.9 and 3.4% of the healthy population (between 800,000 and 3 million persons) have a resting PaO(2) <55 torr, a criterion frequently used for prescribing chronic oxygen therapy in patients with lung diseases. CONCLUSIONS; Although the exact prevalence of hypoxemia in Mexico awaits empirical data, a large number of people live in places where altitude may expose them to low partial pressure of oxygen in inspired air. Ensuing hypoxemia may adversely affect their health. Hypoxia may be particularly harmful to elderly persons and to patients suffering from respiratory diseases. PMID- 11886717 TI - Quantification of vena cava blood flow with half Fourier echo-planar imaging. AB - BACKGROUND: Human hemodynamics occurs in very short periods of time. To quantify blood flow under these circumstances, a fast-scan imaging technique is required. Echo-planar imaging can be a good candidate because it is able to acquire images in <50-100 msec. An imaging scheme with these properties can produce real-time images as well as overcome motion artifacts such as blurring and ghosting, which alter image quality. Additionally, echo-planar imaging does not require a calibration protocol to perform flow experiments in the human cardiovascular system. Consequently, echo-planar imaging appears to be the best imaging tool available to quantify blood flow in the vena cava. From a clinical point of view, echo-planar imaging has become a widespread commodity to produce magnetic resonance images in real-time. METHODS: Flow-encoded half Fourier echo-planar imaging is proposed to determine blood flow in the arteries. This flow sequence was used to investigate vena cava blood flow in healthy volunteers and compared with other diagnostic imaging modalities. RESULTS: Two-dimensional flow maps were obtained by using the two components (sine and cosine images) resulting from the flow-encoded echo-planar imaging sequence. Velocity profiles of vena cava of two healthy volunteers were calculated from the previous bidimensional blood flow maps. CONCLUSIONS: We proved that real-time flow imaging of the cardiovascular system can be achieved with flow-encoded echo-planar imaging and a partial Fourier method. It is possible to quantify blood flow in the superior vena cava in humans. We believe that this imaging tool might offer relevant anatomic and physiologic information of the vena cava as well as of the cardiovascular system. PMID- 11886718 TI - Anti-cardiolipin, anti-cardiolipin plus bovine, or human beta(2)glycoprotein-I and anti-human beta(2)glycoprotein-I antibodies in a healthy infant population. AB - BACKGROUND: Pathogenic antiphospholipid antibodies studies are usually conducted on populations of adults. Studies involving normal children are scant. METHODS: Antibody reactivity against CL alone (true aCL), CL-complexed to bovine beta(2)GP I (aCL-bovine beta(2)GP-I), or human (aCL-human beta(2)GP-I) beta(2)GP-I, or to phospholipid-free human beta(2)GP-I (anti-human beta(2)GP-I) was determined by ELISA in serum samples from 360 Mexican children ranging from 1 month through 8 years of age. RESULTS: Statistical analysis of variance and rankings of Kruskal Wallis demonstrated no significant difference in all tested antibody activities between ages and genders of the study population. Values are presented as a percentile distribution included between 5 and 99, corresponding to the percentages of the studied population. Normal arbitrary units (AU) for IgG, IgA, and IgM true aCL that correspond to the 95 and 99 percentiles are as follows: 2.15 and 3.5; 2.35 and 5.0, and 3.15 and 4.5, respectively. IgG, IgA, and IgM aCL bovine beta(2)GP-I activities are 2.6 and 5.0, 3.0 and 5.0, and 2.7 and 6.0 AU, respectively, while IgG activities of aCL-bovine and human beta(2)GP-I are 1.45 and 1.80, respectively. Normal values for IgG anti-human beta(2)GP-I are 1.85 AU. CONCLUSIONS: While elevated serum levels of antibodies to CL and/or beta(2)GP-I have been associated with thrombotic and hematologic manifestations, the majority of reports deal with adult populations. We report the cut-off values (in AU, international PL units, and international units for beta(2)GP-I) of the specific serologic response of true aCL, aCL-bovine beta(2)GP-I, aCL-human beta(2)GP-I, and anti-human beta(2)GP-I in healthy Mexican children. PMID- 11886719 TI - Frequency and determinants of vitamin A deficiency in children under 5 years of age with pneumonia. AB - BACKGROUND: Vitamin A deficiency (VAD) has been closely related to acute respiratory infections (ARI), although information is still incomplete; for example, the frequency of VAD in children <5 years of age with pneumonia is not known, and the conditions associated with VAD have not been identified. This study was conducted to gain insight into the status of vitamin A in children with pneumonia. A secondary objective was to identify the sociodemographic, individual, and nutritional factors associated with VAD in these children. METHODS: A cross-sectional study was conducted in the Mexican state of Hidalgo, one of the poorest in the country. Children with community-acquired pneumonia treated at nine public hospitals were included. Information was obtained by interviewing mothers, and ascertainment of vitamin A status was performed with relative-dose-response (RDR) test. RESULTS: A total of 422 cases were included. VAD was identified in 17.8% of children; 50.3% showed normal results, 24.6% had liver reserve depletion, and 7.3% showed results attributable to the infectious process. Variables associated with VAD were as follows: age <2 months (OR 3.44, 95% CI: 1.84-9.24); children >6 months of age fed with formula (OR 0.37, 95% CI: 0.15-0.91), and affiliation with the Mexican Social Security Institute (IMSS) health system (OR 0.40, 95% CI: 0.22-0.72). CONCLUSIONS: The frequency of VAD in children with community-acquired pneumonia confirms that the problem of deficiency persists in Mexico. The associated factors for VAD found in this study can be taken into account when planning and evaluating vitamin A supplementation activities in populations with high risk for deficiency. PMID- 11886720 TI - The positive experience of screening quality among users of a cervical cancer detection center. AB - BACKGROUND: Our objective was to determine the main factors associated with increased utilization of a cervical cancer screening program (CCSP) in a population with a high mortality rate due to cervical cancer. METHODS: A population-based study was carried out in the Mexican state of Morelos, Mexico. The study population included 3,197 women between the ages of 15 and 49 years who were selected at random using a State Household Sampling Framework in the State of Morelos's 33 municipalities. The sample included 2,094 women with a history of a previous Papanicolaou (Pap) test. RESULTS: A previous experience of good screening quality is strongly associated with greater use of the CCSP (OR = 4.2; 95% confidence interval [CI], 1.6-10.9). The educational level of the head of the family is related to more frequent use of Pap smear services. Women whose husbands have 13 or more years of education (OR = 1.8; 95% CI 1.1-2.9) were more likely to have been screened. Similarly, women who had used two or more family planning methods (OR = 1.6; 95% CI 1.2-2.1) and those who knew why the Pap test was given (OR = 3.0; 95% CI 2.1-4.3) had a better history of Pap screening. CONCLUSIONS: In areas where coverage of cervical cancer screening is low, a CCSP that guarantees the quality of all the different elements of care is essential if obstacles to cervical cancer prevention are to be eliminated. It is of particular importance to take into account and satisfy the perceptions and expectations of the women at risk. PMID- 11886721 TI - Recurrent epistaxis from Kiesselbach area syndrome in patients suffering from hemorrhoids: fact or fiction? AB - BACKGROUND: It has been found that >90% of patients suffering from recurrent epistaxis from Kiesselbach area syndrome (REKAS) simultaneously suffered from hemorrhoids. To clarify this, the authors decided to investigate in the opposite direction, i.e., to find out whether or not REKAS occurs in patients suffering primarily from hemorrhoids. METHODS: The study group included 53 randomly selected hospitalized patients with hemorrhoidal disorder (31 males and 22 females: age range 18-57 years). A search for essential clinical signs of REKAS was performed in each patient. RESULTS: Incidence was not high, although all clinical parameters were nearly the same: dilated vessels in Kiesselbach venous plexus (83.01%) and a positive hereditary factor (92.7%). The only missing factor in patients with hemorrhoids was anterior septal deformity, so frequent in REKAS patients. CONCLUSIONS; The authors conclude that REKAS and hemorrhoidal syndrome are separate clinical entities that are characterized by dilated vessels of similar venous plexus and simultaneous appearance in the same patient or close relatives. PMID- 11886722 TI - Medium-sized arterial vasculitis associated with vascular deposits of immunoglobin E. Favorable response to intravenous methylprednisolone and cyclophosphamide. AB - Herein we describe the case of a female patient with medium-sized arterial vasculitis associated with high serum immunoglobin E (lgE) levels in the absence of eosinophilia. During occlusive vascular events, IgE deposits in the arterial muscle layer were demonstrated by immunohistochemistry, suggesting a pathogenic role of this antibody in blood vessel inflammation. Combined treatment with methylprednisolone and cyclophosphamide was successful in reducing serum IgE levels and IgE tissue deposits and in inducing the clinical manifestations of vasculitis into remission. This primary medium-sized IgE-associated vasculitis may constitute a new syndrome not previously reported. PMID- 11886723 TI - The advantages of concurrent chemoradiation after neoadjuvant chemotherapy for locally advanced cervical carcinoma. PMID- 11886724 TI - Research with children. PMID- 11886727 TI - James Blundell: the first transfusion of human blood. PMID- 11886728 TI - The epidemiology of out-of-hospital 'sudden' cardiac arrest. AB - It is difficult to assemble data from an out-of-hospital cardiac arrest since there is often lack of objective information. The true incidence of sudden cardiac death out-of-hospital is not known since far from all of these patients are attended by emergency medical services. The incidence of out-of-hospital cardiac arrest increases with age and is more common among men. Among patients who die, the probability of having a fatal event outside hospital decreases with age; i. e. younger patients tend to more often die unexpectedly and outside hospital. Among the different initial arrhythmias, ventricular fibrillation is the most common among patients with cardiac aetiology. The true distribution of initial arrhythmias is not known since several minutes most often elapse between collapse and rhythm assessment. Most patients with out-of-hospital cardiac arrest have a cardiac aetiology. Out-of-hospital cardiac arrests most frequently occur in the patient's home, but the prognosis is shown to be better when they occur in a public place. Witnessed arrest, ventricular fibrillation as initial arrhythmia and cardiopulmonary resuscitation are important predictors for immediate survival. In the long-term perspective, cardiac arrest in connection with acute myocardial infarction, high left ventricular ejection fraction, moderate age, absence of other heart failure signs and no history of myocardial infarction promotes better prognosis. Still there is much to learn about time trends, the influence of patient characteristics, comorbidity and hospital treatment among patients with prehospital cardiac arrest. PMID- 11886729 TI - Impact of age, submersion time and water temperature on outcome in near-drowning. AB - BACKGROUND: Because children have less subcutaneous fat, and a higher surface area to body weight ratio than adults, it has been suggested that children cool more rapidly during submersion, and therefore have a better outcome following near-drowning incidents. AIM OF THE STUDY: To study the impact of age, submersion time, water temperature and rectal temperature in the emergency room on outcome in near-drowning. MATERIAL AND METHODS: This retrospective study included all near-drowning victims admitted to the intensive care units of Helsinki University Central Hospital after successful cardiopulmonary resuscitation between 1985 and 1997. RESULTS: There were 61 near-drowning victims (age range: 0.5-60 years, median 29 years). Males were in the majority (40), and 26 were children (<16 years). The median water temperature was 17 degrees C (range: 0-33 degrees C). The median submersion time for the 43 survivors (70%) was 10 min (range: 1-38 min). Intact survivors and those with mild neurological disability (n=26, 43%) had a median submersion time of 5 min (range: 1-21 min). In non-survivors the median submersion time was 16 min (range: 2-75 min). Submersion time was the only independent predictor of survival in linear regression analysis (P<0.01). Patient age, water temperature and rectal temperature in the emergency room were not significant predictors of survival. CONCLUSIONS: Although submersion time is usually an estimate, it is the best prognostic factor after a near drowning incident. Children did not have a better outcome than adults. PMID- 11886730 TI - Cold water submersion and cardiac arrest in treatment of severe hypothermia with cardiopulmonary bypass. AB - In the paediatric population, submersion injury with drowning or near-drowning represents a significant cause of morbidity and mortality. This study reviews retrospectively our own experiences and the literature on the use of cardiopulmonary bypass (CPB) to rewarm paediatric victims of cold water submersion who suffer severe hypothermia (<28 degrees C) and cardiac arrest (asystole or ventricular fibrillation). In addition to three children treated at our institution, nine other victims were found in the literature. In this cohort of 12 children aged between 2 and 12 years, there was a tendency to better outcome with lower core temperature at the beginning of extracorporeal circulation (mean temperature in nine survivors, 20 degrees C; in three non survivors, 25.5 degrees C). The lowest temperature survived was 16 degrees C. Neither base excess, pH nor serum potassium levels were reliable prognostic factors. The lowest base excess in a survivor was -36.5 mmol/l, the lowest pH 6.29. We consider CPB as the method of choice for resuscitation and rewarming of children with severe accidental hypothermia and cardiac arrest (asystole or ventricular fibrillation). Compared with adults, children, especially smaller ones, require special consideration with regard to intravenous cannulation as drainage can be inadequate using femoral-femoral cannulation. In hypothermic children we advocate, therefore, emergency median sternotomy. Until more information regarding prognostic factors are available, children who are severely hypothermic and clinically dead after submersion in cold water--even if for an unknown length of time--should receive cardiopulmonary resuscitation (CPR) and be transported without delay to a facility with capabilities for CPB instituted via a median sternotomy. PMID- 11886731 TI - Should we follow ATLS guidelines for the management of traumatic pulmonary contusion: the role of non-invasive ventilatory support. AB - OBJECTIVE: To assess the management of patients with blunt traumatic pulmonary contusion admitted to our hospital. To identify the role of early blood gas analysis, non-invasive ventilation and to assess the validity of the current Advanced Trauma Life Support manual statement that "Patients with significant hypoxia, i.e. PaO(2)<65 mm Hg or 8.6 kPa on room air, SaO(2)<90%, should be intubated and ventilated within the first hour after injury". SETTING: A 24 bed Intensive Care Unit in a major Trauma Centre situated in South Western Sydney, Australia. METHODS: Retrospective review of adults with blunt traumatic pulmonary contusion identified from the trauma registry. RESULTS: A total of 75 patients with an age range of 16-81 years were identified over a 2-year period. Arterial blood gas measurement was available for 32 patients during the immediate resuscitative period (<1 h from admission). All patients received supplemental oxygen and a PaO(2)/FiO(2) ratio was calculated. Seven patients had significant pulmonary contusion, indicated by an initial PaO(2)/FiO(2) ratio of <300, and were treated successfully with non-invasive ventilatory support. A further five patients without arterial blood gas (ABG) analysis on admission but with PaO(2)/FiO(2) ratio of <300 in the ICU were also managed with non-invasive ventilatory support. Multi-modal analgesia was commonly used. CONCLUSIONS: All major trauma patients admitted to our hospital received supplemental oxygen. Interpretation of ABG breathing room air was not used as an indicator for intubation. Most decisions to intubate early were based on clinical need. Patients with significant pulmonary contusion required intubation for reasons other than respiratory failure. Patients with significant pulmonary contusion were managed safely with non-invasive ventilatory support. Further investigation will determine the role of non-invasive ventilatory support in the management of these patients. PMID- 11886732 TI - Survival to discharge following open chest cardiac compression (OCCC). A 4-year retrospective audit in a cardiothoracic specialist centre--Royal Brompton and Harefield NHS Trust, United Kingdom. AB - OBJECTIVE: To review the use of Open Chest Cardiac Compression (OCCC) techniques in postcardiac surgical patients in one specialist cardiothoracic centre in the UK. METHODS: A 4-year retrospective audit (April 1995--March 1999) of all cardiac arrest victims and resuscitation practice across two specialist cardiothoracic hospitals. Audit outcomes related to initial survival and survival to discharge, arrest rhythm, reasons for resternotomy, surgical procedure prior to resternotomy and time elapsed from original surgery to resternotomy. RESULTS: Seventy-two patients (adult and paediatric) suffering cardiac arrest received OCCC following cardiac surgery. Thirty-three patients initially survived (46%) and 12 patients survived to discharge (17%). DISCUSSION AND RECOMMENDATIONS: In the absence of current European Resuscitation Council guidelines, we adopted recommendations for resternotomy to be performed after 5 min of unsuccessful conventional CPR and OCCC initiated. An adapted ERC algorithm incorporating these recommendations can provide much needed direction in postcardiac surgery cardiac arrest victims. PMID- 11886734 TI - ALERT--a multiprofessional training course in the care of the acutely ill adult patient. AB - The Acute Life-threatening Events--Recognition and Treatment (ALERT) course is a one-day multidisciplinary course originally designed to give newly qualified doctors and nurses greater confidence and ability in the recognition and management of adult patients who have impending or established critical illness. It may also be suitable for many other groups of health service workers. ALERT was developed using principles common to many advanced life support courses and incorporates aspects of clinical governance, multidisciplinary education and interprofessional working. It incorporates pre-course reading, informal and interactive seminars, practical demonstrations and role-play during clinically based scenarios. A novel aspect of ALERT is that participants undertake role interchange during scenarios, thereby facilitating mutual understanding. At all times during the course, participants are encouraged to reflect on their actions and to pay particular attention to detail. The course focuses on those problems that lead ward nurses to call doctors for assistance, e.g. 'the blue patient', 'the hypotensive patient'. Communication skills are covered frequently in the course, during seminars and scenarios, but also as a specific session that covers three aspects--breaking bad news, writing patient notes and interpersonal/interprofessional communication. PMID- 11886733 TI - Retention of basic life support skills 6 months after training with an automated voice advisory manikin system without instructor involvement. AB - AIM: To evaluate the retention of skills 6 months after training in ventilation and chest compressions (CPR) on a manikin with computer based on-line voice advisory feedback and the possible effects of initial overtraining. METHODS: Thirty five volunteers had 20 min provisional CPR training on a manikin with computer based voice advisory feedback but without an instructor. The appropriate feedback was taken from a pre-recorded list depending on performance measured by the manikin--computer system versus set limits for ventilation and compression variables. One group in addition was randomised to receive 10 similar 3 min training sessions during 1 week in the following month (overtrained group). All ventilation and compression variables were measured without feedback before and after the initial training session, with feedback immediately thereafter, and both without and with feedback 6 months after the initial training session. RESULTS: The initial training improved all variables. Compressions with correct depth increased from a mean of 33 to 77%, and correct inflations from a mean of 9 to 58%. After 6 months, the results for the controls were not significantly different from pre-training, except for a higher of correct inflations (18%), while the overtrained group had better retention of skills including the correct compression depth (mean 61%) and inflations (mean 42%). When verbal feedback was added both the compressions and ventilations immediately improved both when tested immediately and 6 months after the initial training session. CONCLUSIONS: The computer-based voice advisory manikin (VAM) feedback system can improve immediate performance of basic life support (BLS) skills, with better long-term retention with overtraining. PMID- 11886735 TI - Teaching basic life support skills using self-directed learning, a self instructional video, access to practice manikins and learning in pairs. AB - Applying adult learning principles in healthcare education is increasingly recognised as useful and effective. We designed and evaluated an educational package for medical student basic life support (BLS) skills that placed the responsibility of skill acquisition with the learner. The package provided hardcopy and web based information, an in-house produced audio-video tape demonstrating BLS, and open access to manikins in a Skills Centre where the students learnt in pairs. Students determined when they were ready to be assessed. This assessment was performed by two independent observers using the Resuscitation Council (UK) BLS assessment sheet. Two groups, comprising in total 51 fourth year medical students were assessed, 47 were found to be competent in performing BLS on their first assessment. Of the remaining four, three were assessed as competent after further self-directed learning and retesting. Only one student required personal tutoring prior to success. Self-directed learning is a successful method of mastering BLS. Where failure occurred, it was due to inadequate student learning in the Skills Centre. The importance of practice needs emphasis in future use of the programme, as does the virtual guarantee of success, if all steps are followed. A similar programme could be devised for other technical skills. PMID- 11886736 TI - Dashing with scooters to in-hospital emergencies: a randomised cross-over experiment. AB - OBJECTIVE: Physical exhaustion is a frequent condition in emergency medical teams after in-house emergency runs, which might affect the quality of advanced care. Newly available light-weight scooters may reduce exertion as measured by the cardiovascular response in these circumstances and, therefore, may reduce physical exhaustion on arrival. METHODS: We undertook a randomised cross-over trial in a simulated in-house emergency alarm run to examine the influence of scooting compared with conventional running on pulse rate (primary outcome), manual skillfulness and response time. RESULTS: We tested 24 emergency department professionals in eight emergency medical teams. After scooting the pulse rate was significantly lower compared with conventional running [157 (IQR 145-169) vs. 170 (IQR 154-175) min(-1), P=0.004]. After the simulated emergency alarm run no difference was found in manual skillfulness and response time between scooting and running. CONCLUSION: Using scooters for simulated in-house emergency alarm runs markedly reduces the cardiovascular response of emergency medical teams. PMID- 11886737 TI - Use of heliox in critical upper airway obstruction. Physical and physiologic considerations in choosing the optimal helium:oxygen mix. AB - Heliox has a lower density than oxygen and nitrogen, and can improve ventilation rapidly in patients with critical upper airway obstruction. The choice of the best helium:oxygen ratio depends on whether the predominant problem is hypercarbia or hypoxia. In the former situation, 80% helium should be used, and in the latter, 100% oxygen is appropriate. PMID- 11886739 TI - ILCOR/ERC recommendations for out-of-hospital 12-lead ECG and fibrinolysis are not well adopted at present. PMID- 11886738 TI - Hurrah--we are still alive! A different dimension in post-resuscitative care: the annual gathering of cardiac arrest survivors at a typical Viennese wine tavern. PMID- 11886740 TI - Electrocardiographic changes in occult pneumothorax. PMID- 11886741 TI - Thrombolysis using recombinant tissue-type plasminogen activator during cardiopulmonary resuscitation in patients with out-of-hospital cardiac arrest. PMID- 11886742 TI - Direct detection of N-acylhomoserine lactones in cystic fibrosis sputum. AB - Pseudomonas aeruginosa and Burkholderia cepacia cause destructive lung disease in cystic fibrosis (CF) patients. Both pathogens employ 'quorum sensing', i.e. cell to-cell communication, via diffusible N-acyl-L-homoserine lactone (AHL) signal molecules, to regulate the production of a number of virulence determinants in vitro. However, to date, evidence that quorum sensing systems are functional and play a role in vivo is lacking. This study presents the first direct evidence for the presence of AHLs in CF sputum. A total of 42 samples from 25 CF patients were analysed using lux-based Escherichia coli AHL biosensors. AHLs were detected in sputum from patients colonised by P. aeruginosa or B. cepacia but not Staphylococcus aureus. Furthermore, using liquid chromatography-mass spectrometry and thin layer chromatography, we confirmed the presence of N-hexanoylhomoserine lactone and N-(3-oxododecanoyl)homoserine lactone respectively in sputum samples from patients colonised by P. aeruginosa. PMID- 11886743 TI - Protein aggregation in Escherichia coli: role of proteases. AB - Protein aggregation is involved in several human diseases, and presumed to be an important process in protein quality control. In bacteria, aggregation of proteins occurs during stress conditions, such as heat shock. We studied the protein aggregates of Escherichia coli during heat shock. Our results demonstrate that the concentration and diversity of proteins in the aggregates depend on the availability of proteases. Aggregates obtained from mutants in the Lon (La) protease contain three times more protein than wild-type aggregates and show the broadest protein diversity. The results support the assumption that protein aggregates are formed from partially unfolded proteins that were not refolded by chaperones or degraded by proteases. PMID- 11886744 TI - Characterization of the isoprenoid chain of coenzyme Q in Plasmodium falciparum. AB - Little is known about isoprenoid biosynthesis in parasitic protozoa. The presence of dolichol and isoprenylated proteins has been detected in Plasmodium falciparum, but no studies are available about the biosynthesis of the isoprenic side chain attached to the benzoquinone ring of coenzyme Q. In the present study, using metabolic labelling with different intermediates, we demonstrated the presence of an active isoprenoid pathway for the biosynthesis of the isoprenic chain of coenzyme Q. Our results show that P. falciparum is able to synthesize different homologs (coenzyme Q(8) and coenzyme Q(9)), depending on the given intermediate. Parasites treated with nerolidol at doses 2.2 times below the IC(50) showed a decreased ability to synthesize the isoprenic chain attached to coenzyme Q at all intraerythrocytic stages. Treatment with nerolidol arrested development of the intraerythrocytic stages of the parasites, indicating that the drug may have an antimalarial potential. PMID- 11886745 TI - Genetic relationships of Bacillus anthracis and closely related species based on variable-number tandem repeat analysis and BOX-PCR genomic fingerprinting. AB - Variable-number tandem repeats (VNTR) analysis and BOX-repeat-based PCR (BOX-PCR) genomic fingerprinting were performed on 25 Bacillus strains to investigate the genetic relatedness of Bacillus anthracis to the closely related species. Based on VNTR analysis, all B. anthracis strains could be assigned to (VNTR)(4), which is the most commonly found type in the world. Interestingly, a (VNTR)(2) was also observed in Bacillus cereus KCTC 1661 and with an exact match to the tandem repeats found in B. anthracis. This finding has never been reported before in the closely related species. According to the BOX-PCR, B. anthracis strains clustered together and separated reliably from the closely related species. However, B. cereus KCTC 1661 was linked to the B. anthracis cluster and showed close relationships with B. anthracis strains. These results indicated that there was a strong correlation between VNTR analysis and BOX-PCR genomic fingerprinting. PMID- 11886746 TI - Expression of a small RNA, BS203 RNA, from the yocI-yocJ intergenic region of Bacillus subtilis genome. AB - We isolated and characterized a novel small RNA from Bacillus subtilis. We termed this molecule BS203 RNA from the length of its mature form (203 nt) and located the corresponding gene at the yocI-yocJ intergenic region on the B. subtilis genome. Northern blotting revealed that it is transcribed in vegetative growing cells and that the amount of BS203 RNA decreased in the middle of the vegetative phase. A computer-aided prediction of the BS203 RNA secondary structure revealed three characteristic stem-loop structures. Despite active expression during the vegetative phase, growth of the knockout mutant was not affected by depletion of BS203 RNA. A phylogenetic comparison of the sequence of the BS203 RNA with other Bacillus species including B. cereus and B. halodurans C-125, or Clostridium perfringens suggests that the sequence is unique to Bacillus subtilis. PMID- 11886747 TI - Kinetic analysis of PPi-dependent phosphofructokinase from Porphyromonas gingivalis. AB - We have previously cloned the gene encoding a pyrophosphate-dependent phosphofructokinase (PFK), designated PgPFK, from Porphyromonas gingivalis, an oral anaerobic bacterium implicated in advanced periodontal disease. In this study, recombinant PgPFK was purified to homogeneity, and biochemically characterized. The apparent K(m) value for fructose 6-phosphate was 2.2 mM, which was approximately 20 times higher than that for fructose 1,6-bisphosphate. The value was significantly greater than any other described PFKs, except for Amycolatopsis methanolica PFK which is proposed to function as a fructose 1,6 bisphosphatase (FBPase). The PgPFK appears to serves as FBPase in this organism. We postulate that this may lead to the gluconeogenic pathways to synthesize the lipopolysaccharides and/or glycoconjugates essential for cell viability. PMID- 11886748 TI - 23S rRNA point mutation associated with erythromycin resistance in Treponema denticola. AB - Mechanisms and occurrence of macrolide resistance in the periodontal pathogen Treponema denticola have received little attention. In this study, erythromycin resistance due to mutations in the genes encoding T. denticola 23S rRNA was investigated. The T. denticola genome was shown to contain two copies of 23S rDNA. 23S rRNA genes of nine erythromycin-resistant isolates derived from T. denticola were amplified and sequences were analyzed. All the erythromycin resistant strains had at least one A-->G transition mutation at the 23S rRNA gene sequence cognate to position A2058 in Escherichia coli 23S rDNA. This suggests that antibiotic pressure is sufficient to select for point mutations that confer resistance in this organism. PMID- 11886749 TI - Sequence analysis of 16S rRNA gene of cyanobacteria associated with the marine sponge Mycale (Carmia) hentscheli. AB - Marine sponges frequently contain a complex mixture of bacteria, fungi, unicellular algae and cyanobacteria. Epifluorescent microscopy showed that Mycale (Carmia) hentscheli contained coccoid cyanobacteria. The 16S rRNA gene was amplified, fragments cloned and analysed using amplified rRNA gene restriction analysis. The nearly complete 16S rRNA gene of distinct clones was sequenced and aligned using ARB. The phylogenetic analysis indicated the presence of four closely related clones which have a high (8%) sequence divergence from known cyanobacteria, Cyanobacterium stanieri being the closest, followed by Prochloron sp. and Synechocystis sp. All belong to the order Chroococcales. The lack of non molecular evidence prevents us from proposing a new genus. PMID- 11886750 TI - Utilisation of aminomethane sulfonate by Chromohalobacter marismortui VH1. AB - Chromohalobacter marismortui VH1 was screened for its ability to utilise organosulfonate compounds at a range of NaCl concentrations. Only aminomethane sulfonate, of seven sulfonates tested, was utilised. Length of lag phase during growth on aminomethane sulfonate, as either nitrogen and/or sulfur source, increased with increasing NaCl concentration. Cell yields increased linearly with increasing aminomethane sulfonate concentration up to 5 mM. Resting cells pregrown on aminomethane sulfonate as sole nitrogen source exhibited carbon sulfur bond cleaving [0.123 nmol sulfate accumulated h(-1) (mg cells)(-1)] and sulfite-oxidising [0.185 nmol sulfate accumulated h(-1) (mg cells)(-1)] activities. C. marismortui VH1 is capable of sulfur-starvation deregulated metabolism of aminomethane sulfonate under high salt conditions. PMID- 11886751 TI - MOSC domains: ancient, predicted sulfur-carrier domains, present in diverse metal sulfur cluster biosynthesis proteins including Molybdenum cofactor sulfurases. AB - Using computational analysis, a novel superfamily of beta-strand-rich domains was identified in the Molybdenum cofactor sulfurase and several other proteins from both prokaryotes and eukaryotes. These MOSC domains contain an absolutely conserved cysteine and occur either as stand-alone forms such as the bacterial YiiM proteins, or fused to other domains such as a NifS-like catalytic domain in Molybdenum cofactor sulfurase. The MOSC domain is predicted to be a sulfur carrier domain that receives sulfur abstracted by the pyridoxal phosphate dependent NifS-like enzymes, on its conserved cysteine, and delivers it for the formation of diverse sulfur-metal clusters. The identification of this domain may clarify the mechanism of biogenesis of various metallo-enzymes including Molybdenum cofactor-containing enzymes that are compromised in human type II xanthinuria. PMID- 11886752 TI - Transcriptional regulation of mexR, the repressor of Pseudomonas aeruginosa mexAB oprM multidrug efflux pump. AB - The transcription start site of mexR, encoding the repressor of the Pseudomonas aeruginosa mexAB-oprM multidrug efflux pump, has been determined by S1 mapping. One signal corresponding to a single promoter has been found, whereas three major signals were observed for the mexA messenger. Further analysis demonstrated that mexA has just one promoter that overlaps with the mexR promoter, with the other two signals observed by S1 probably being the consequence of RNA processing. Transcription of mexR and mexA from the aforementioned promoters is regulated by MexR. We show that bacterial growth phase affects expression of these promoters as well. mexR expression was higher at the exponential growth phase and declined afterwards, whereas mexA expression was triggered at the onset of the stationary growth phase. A model for the regulation of mexR and mexA expression, which includes an analysis of the interplay between both promoters, is proposed. PMID- 11886753 TI - Analysis of the Schwanniomyces occidentalis SWA2 gene promoter in Saccharomyces cerevisiae. AB - The effect of different carbon sources on the expression in Saccharomyces cerevisiae of the SWA2 alpha-amylase gene from Schwanniomyces occidentalis was studied from constructs containing its 5' region (-223 to +15), which were fused in-frame to the lacZ gene coding sequence. Maximal expression was achieved with the non-fermentable substrates ethanol and/or glycerol, whereas lower levels were found with maltose or galactose. In contrast, glucose repressed it, even in the presence of any of these other carbon sources. Deletion analyses of the -233 to 85 SWA2 promoter region permitted the identification of two fragments involved in both glucose repression and ethanol activation. A possible region required for cAMP regulation was localised. The SWA2 promoter contains a MIG1-binding GC box whose deletion caused a five-fold increase in the glucose-repressed reporter expression. Despite this, expression of the SWA2 promoter was not MIG1-dependent. PMID- 11886754 TI - Natural transformation of Pseudomonas stutzeri by single-stranded DNA requires type IV pili, competence state and comA. AB - Pseudomonas stutzeri, in addition to being transformed by duplex DNA, is also transformed by the sense or antisense strand of the genetic marker employed (hisX(+)) or by heat-denatured chromosomal DNA. Transformation was absent in non competent cells and in mutants defective for pilus biogenesis (pilA, pilC) and function (pilT) or DNA translocation into the cytoplasm (comA). Uptake of (3)H thymidine-labeled single-stranded DNA was hardly detectable reflecting the 20- to 60-fold lower transformation compared to duplex DNA. The results suggest that the steps in natural transformation also accommodate single-stranded DNA and that DNA translocation from the periplasm into the cytoplasm is not necessarily coupled to the degradation of a complementary strand. Small DNA single-stranded fragments are thus not excluded from horizontal gene transfer by transformation. PMID- 11886755 TI - Inactivation of the Mycobacterium tuberculosis Nramp orthologue (mntH) does not affect virulence in a mouse model of tuberculosis. AB - Mycobacterium tuberculosis is an intracellular pathogen which can survive and multiply within the phagosomal compartment of the macrophage, and in doing so has to withstand the various macrophage defense mechanisms, which include limitation of iron and other metals. Analysis of the complete genome sequence of M. tuberculosis revealed an extensive array of cation transporters, including mntH, an orthologue of the eukaryotic Nramp (natural resistance-associated macrophage protein) gene, that encodes a proton-dependent divalent metal transporter. To assess the effect of this transporter on intracellular survival and pathogenesis, an mntH knock-out mutant of M. tuberculosis H37Rv was created and assayed in bone marrow-derived macrophages and in a murine model of tuberculosis. In neither of these systems was any loss of fitness associated with inactivation of mntH, demonstrating that Nramp orthologues are not important determinants of mycobacterial virulence. PMID- 11886756 TI - Identification of lactoferrin-binding proteins in Streptococcus dysgalactiae subsp. dysgalactiae and Streptococcus agalactiae isolated from cows with mastitis. AB - Three strains of Streptococcus dysgalactiae subsp. dysgalactiae (S. dysgalactiae) and five strains of Streptococcus agalactiae were used to identify lactoferrin binding proteins (LBPs). LBPs from extracted surface proteins were detected by polyacrylamide gel electrophoresis and Western blotting. All strains of S. dysgalactiae evaluated had 52- and 74-kDa protein bands. All strains of S. agalactiae evaluated had 52-, 70- and 110-kDa protein bands. In addition, a 45 kDa band was detected in two of five S. agalactiae strains evaluated. This study demonstrated that S. dysgalactiae and S. agalactiae of bovine origin contain two and three major LBPs, respectively. PMID- 11886757 TI - Increased expression of the multidrug efflux genes acrAB occurs during slow growth of Escherichia coli. AB - Intrinsic antimicrobial resistance of Escherichia coli is elicited by the gene products of the multidrug efflux acrAB-tolC operon. In this paper, we have shown that acrAB is regulated as a function of the growth rate of E. coli during growth in batch and chemostat culture. In chemostat culture, expression of acrAB is inversely related to growth rate irrespective of the limiting nutrient. The level of expression of acrAB is greater under glucose limitation compared with either iron or nitrogen limitation. Increase in expression of acrAB confers a greater resistance to ciprofloxacin, and the implications for a clinical situation are discussed. Slow growth rate regulation of acrAB transcription does not require the presence of the stationary-phase sigma factor. A putative gearbox consensus sequence was identified at the -10 region of the acrAB promoter. PMID- 11886758 TI - Synthesis of polyhydroxyalkanoate in the peroxisome of Pichia pastoris. AB - Polyhydroxyalkanoates (PHAs) are polyesters naturally produced by bacteria that have properties of biodegradable plastics and elastomers. A PHA synthase from Pseudomonas aeruginosa modified at the carboxy-end for peroxisomal targeting was transformed in Pichia pastoris. The PHA synthase was expressed under the control of the promoter of the P. pastoris acyl-CoA oxidase gene. Synthesis of up to 1% medium-chain-length PHA per g dry weight was dependent on both the expression of the PHA synthase and the presence of oleic acid in the medium. PHA accumulated as inclusions within the peroxisomes. P. pastoris could be used as a model system to study how peroxisomal metabolism needs to be modified to increase PHA production in other eukaryotes, such as plants. PMID- 11886759 TI - DNA sequence and genetic characterization of plasmid pFQ11 from Frankia alni strain CpI1. AB - An 8551-bp plasmid, pFQ11, from Frankia alni strain CpI1 was sequenced. Its sequence was found to be very similar to that presented for pFQ31 from strain ArI3. Six potential protein-encoding open reading frames (ORFs) were identified, and transcriptional activity was shown within four of those regions of the plasmid by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. An earlier study reported that ORF E(F) of pFQ31, which is nearly identical to the 3' 45% of ORF1 of pFQ11, is significantly similar to RepF. We found no such similarity. ORF2 and ORF3 predict products that are similar to a repressor protein and a partition protein, respectively. We found inverted repeats within and covering the start codon of ORF3; palindromic sequences and direct repeats between ORF3 and ORF4; and 3' from ORF3, an AT-rich sequence that extensively overlaps the promoter region of a uvrB homolog in strain ArI3. PMID- 11886761 TI - Our history. PMID- 11886762 TI - The meaning of mammographic breast density in users of postmenopausal hormone therapy. PMID- 11886763 TI - Lipoproteins and BMI: a comparison between women during transition to menopause and regularly menstruating healthy women. AB - One hundred and forty-three women born 1942 were followed for 5 years during transition to menopause (49--54 years of age). Changes in menopausal status, body mass index (BMI) and circulating lipoproteins cholesterol, (chol), low density lipoprotein (LDL), high density lipoprotein, (HDL) and total triglycerides (TG) were measured, once yearly and compared with a control group of normally menstruating healthy, non-smoking women 23--39 years old. RESULTS: Chol was significantly higher P<0.0001 in the study group visits 1--5 when compared with the controls and higher at visit 4 compared with visit 1(P<0.05) LDL was significantly lower in the study group and at visit 5 compared with visit 2 (P<0.05) HDL was significantly lower at visits 1--3 and 5 when compared with the controls (P<0.001) and to visit 4 (P<0.0001). TG was significantly higher in the study group (P<0.0001) and increased significantly during the 5-year study. BMI was significantly higher in the older women (P<0.001) and increased during 5 years of study (P<0.0001). When multiple stepwise regression analyses were performed at visit 5 using education, menopausal status, use of hormone replacement therapy (HRT), BMI and smoking as predictor variables, postmenopausal status was found to be significantly associated with high LDL (P<0.3), while high BMI significantly predicted low HDL and high TG levels. Perimenopausal status was significantly associated with high HDL levels. CONCLUSION: Age, BMI and menopausal status are significant predictors of circulating lipoprotein levels during transition to menopause. PMID- 11886764 TI - Cytokine pattern in postmenopause. AB - OBJECTIVE: The present study was undertaken to evaluate the pattern of serum cytokine production in postmenopausal women and the relationship with the hormonal status. A group of fertile women served as controls. METHODS: Eighty-two women in apparent good health, non-smokers and without a history of hormone replacement therapy, were enrolled for the study. The women were divided in two groups according to their hormonal status: fertile women (n=34, age 32 +/- 7 years) and postmenopausal women (n=48, age 54 +/- 8 years). Blood samples were withdrawn in the morning, after an overnight fast. RESULTS: Sex hormones (LH, FSH, Estradiol, Progesterone, DHEA, DHEA-S), as well as GH and IGF-1 levels, were significantly higher in the serum of fertile women as compared with their postmenopausal counterparts. Unlike IL-2, IL-4, IL-5, IL-10, IL-12 and IFN-gamma, significant differences were observed in serum IL-6, IL-18, TNFalpha and TNFbeta between groups: both IL-6 and IL-18 were higher in postmenopausal women, while TNFalpha and TNFbeta were significantly lower. There was an inverse relationship between serum DHEA and DHEA-S levels and both IL-6 (r= -0.46, P<0.02) or IL-18 (r= -0.38, P<0.05) serum concentrations. CONCLUSIONS: Compared with fertile counterparts, women in postmenopause present an alteration in serum cytokine profile suggesting a prevalence of Th2 lymphocytes. PMID- 11886765 TI - Estrogen receptor alpha polymorphism as a genetic marker for bone loss, vertebral fractures and susceptibility to estrogen. AB - OBJECTIVE: The purpose of the present study was to investigate the possible roles of PvuII and XbaI polymorphisms of the estrogen receptor alpha (ER(alpha)) in bone mineral density (BMD), vertebral fracture, bone loss rate after menopause and response to hormone replacement therapy (HRT). METHODS: All 286 women were grouped according to the genotypes of PvuII or XbaI polymorphisms of the ER(alpha) gene. We compared the BMD Z-score, incidence of vertebral fracture, changes in Z-score after menopause and response of BMD to HRT among the genotypes. RESULTS: Subjects with the PPxx genotype had significantly (P<0.05) lower Z-scores than did subjects with the other genotypes. A negative correlation was observed between the length of time after menopause and the decrease of the Z score only in women with the pp genotype, suggesting faster bone loss in this group. In the analysis of the ER(alpha) polymorphism with regard to the effect of HRT on BMD, there appears to be a significantly greater increase of BMD (P<0.01 and 0.05) in women with the pp genotype than in those with the Pp or PP genotype. CONCLUSIONS: PvuII and XbaI polymorphisms of the ER(alpha) gene were associated with BMD in postmenopausal Japanese women. Also, the polymorphisms may be useful genetic markers for predicting vertebral fracture in relatively young postmenopausal women. The PvuII polymorphism may be associated with susceptibility to changes in estrogen level. PMID- 11886766 TI - Cross-sectional study of the effects of parturition and lactation on bone mineral density later in life. AB - OBJECTIVE: The present cross-sectional study investigated the effects of parturition and lactation on bone mineral density (BMD) later in life. METHODS: The subjects were 456 premenopausal and 713 postmenopausal Japanese women aged 40 69 years old. They were classified into six subgroups at 5-year increments. Age, height, weight, menopausal status, age at menopause (in postmenopausal women), years since menopause (in postmenopausal women), parity, and total lactation period were recorded. Lumbar spine BMD (L2-4) was measured by dual-energy X-ray absorptiometry (DEXA). In each subgroup, correlations of parturition and lactation with BMD were investigated using Pearson's correlation test and multiple regression analyses. RESULTS: In premenopausal women aged 40-44 years old (n=143), total lactation period was inversely correlated with BMD (r= -0.293, P<0.01). This relationship remained significant after adjusting for age, height, weight, and parity (P<0.05). Although the total lactation period was inversely correlated with BMD in the group aged 60-64 years old (r= -0.194, P<0.05, n=218), this relationship disappeared after adjusting for age, YSM, height, weight, and parity. However, in the other subgroups, there were no significant correlations between total lactation period and BMD. There were no significant correlations observed between parity and BMD in any groups. CONCLUSION: Reproductive history of lactation and parity does not seem to be a major determinant of BMD later in life. PMID- 11886767 TI - Effect of continuous combined therapy with vitamin K(2) and vitamin D(3) on bone mineral density and coagulofibrinolysis function in postmenopausal women. AB - OBJECTIVES: To investigate the therapeutic effect of combined use of vitamin K(2) and D(3) on vertebral bone mineral density in postmenopausal women with osteopenia and osteoporosis. SUBJECTS AND METHODS: We enrolled 172 women with vertebral bone mineral density <0.98 g/cm(2) (osteopenia and osteoporosis) as measured by dual-energy X-ray absorptiometry. In this study, we employed the criteria for diagnosis of osteopenia and osteoporosis using dual energy X-ray absorptiometry proposed by the Japan Society of Bone Metabolism in 1996. Subjects were randomized into four groups (each having 43 subjects in vitamin K(2) therapy group, vitamin D(3) therapy group, vitamin K(2) and D(3) combined therapy group, or a control group receiving dietary therapy alone) and treated with respective agents for 2 years, with bone mineral density was measured prior to therapy and after 6, 12, 18, and 24 months of treatment. The bone metabolism markers analyzed were serum type 1 collagen carboxyterminal propeptide (P1CP), serum intact osteocalcin, and urinary pyridinoline. Tests of blood coagulation function consisted of measurement of activated partial thromboplastin time (APTT) and analysis of concentrations of antithrombin III (AT III), fibrinogen, and plasminogen. RESULTS: Combined therapy with vitamin K(2) and D(3) for 24 months markedly increased bone mineral density (4.92 +/- 7.89%), while vitamin K(2) alone increased it only 0.135 +/- 5.44%. The bone markers measured, revealed stimulation of both bone formation and resorption activity. We observed an increase in coagulation and fibrinolytic activity that was within the normal range, suggesting that balance was maintained in the fibrinolysis-coagulation system. CONCLUSIONS: Continuous combination therapy with vitamin K(2) and D(3) may be useful for increasing vertebral bone mass in postmenopausal women. Furthermore, the increase in coagulation function observed during this therapy was within the physiological range, and no adverse reactions were observed. PMID- 11886768 TI - Resistive index of renal artery and blood pressure in postmenopausal women. AB - Causal association between perimenopausal changes and symptoms and disease is commonly accepted even if not definitely explained. Resistive index (RI) of renal artery assessed by Doppler echography is related to renal function and systemic circulatory adaptation in patients with chronic renal failure and hypertension. Echocardiographic measurement of left ventricular myocardial mass (LVMM) is a useful tool for assessing effects of arterial hypertension on heart. Aim of the study was to assess RI in normotensive postmenopausal women and relationship, if any, with blood pressure and LVMM. We studied 28 normotensive, non-obese postmenopausal women, age 52.21 +/- 5.40 years, with normal creatinine clearance. Renal colour-Doppler echography was performed assessing intra-parenchimal renal artery mean velocity (mVRA) and intra-parenchimal RI [(peak systolic velocity - end diastolic velocity)/peak systolic velocity]. Echocardiography was performed as well. RI of intra-parenchimal renal artery is 0.67 +/- 0.05 and it shows correlations vs. diastolic blood pressure (r=0.41; P<0.03) and vs. mean BP (r=0.47; P<0.01). LVMM has correlation (r=0.41; P>0.03) with RI. Age, body weight, body mass index, menarche age, fertility years and postmenopausal years do not show correlation with RI. Heart rate, creatinine clearance, hemoglobin, serum albumin do not show any correlation with RI. Higher RI is associated with alcohol intake, liver steatosis, biliary gallstones and family history of diabetes mellitus, but not with postmenopausal years, unrespective of surgical or non-surgical menopause. Among echocardiographic measurements only LVMM is correlated with RI; mVRA does not show correlation. LVMM and BP do not show other independent correlation except that the one already reported vs. RI. RI, as a pathophysiological measurement whose increase preludes to arterial hypertension, could help to ascertain perimenopausal women at risk for arterial hypertension and left ventricular hypertrophy, but does not seem directly associated with the loss of ovarian function. PMID- 11886769 TI - Efficacy of a new 7-day transdermal sequential estradiol/levonorgestrel patch in women. AB - OBJECTIVE: To investigate the efficacy and tolerability of a new 7-day transdermal sequential estradiol/levonorgestrel patch (Fem7 Combi; Merck KGaA; Germany), versus placebo, as hormone replacement therapy in menopausal women. METHODS: A multicentre, randomized, clinical study consisting of a 3-week screening phase, a 12-week double-blind, placebo-controlled treatment phase, and a 12-week open, follow-up phase. Women aged 40-65 years with an intact uterus and menopausal complaints were randomized to either 2 weeks of an estradiol mono patch (50 microg per 24 h) followed by 2 weeks of an estradiol/levonorgestrel combination patch (50 microg/10 microg per 24 h), or a placebo patch, for three 28-day cycles. Changes in the Kupperman Index and the frequency of hot flushes were assessed. RESULTS: The sequential use of a 7-day estradiol patch and a 7-day estradiol/levonorgestrel patch was superior to placebo in reducing menopausal symptoms, and was well tolerated. At the end of the treatment phase, there was a statistically significant reduction in the Kupperman Index score versus placebo (P<0.0001), and a statistically significant difference between groups in the proportion of patients with a reduction in the number of hot flushes (at least 50% versus baseline). During the open follow-up phase, there was a marked reduction in the Kupperman Index score and the number of hot flushes for patients switched from placebo to active study medication. The active medication was effective throughout the 1-week application period. CONCLUSIONS: The new 7-day transdermal sequential estradiol/levonorgestrel patch was well tolerated, providing rapid and effective relief of menopausal symptoms. The addition of low dose levonorgestrel did not influence the beneficial effects of estradiol. PMID- 11886770 TI - Rhodnius heme-binding protein (RHBP) is a heme source for embryonic development in the blood-sucking bug Rhodnius prolixus (Hemiptera, Reduviidae). AB - We have previously shown that Rhodnius prolixus' eggs and hemolymph are pink due to the presence of the hemeprotein Rhodnius heme-binding protein (RHBP). In the hemolymph it functions as an antioxidant. Nevertheless, its function in eggs has not been determined. Here we present evidence that RHBP is a source of heme for embryonic development. RHBP content decreases during embryogenesis, but the total heme content of eggs remains unchanged. Biliverdin, the product of heme degradation, is not detectable in late embryos. The activity of the heme synthesizing pathway is low throughout embryogenesis and rises sharply after nymphs' hatching. Heme-radiolabeled eggs were produced and, at the day of hatching, nymphs were dissected. The presence of radiolabeled heme in their carcass is an indication that heme reutilization is occurring. The only animal known to reutilize heme in significant levels is the cattle tick Boophilus microplus, which cannot synthesize its own heme. Diversely, Rhodnius can synthesize its own heme but, in the context of embryogenesis, heme demand seems to be supplied by the programmed release of heme form RHBP. This behavior indicates that in Rhodnius, we might have a highly unusual profile: heme is both synthesized and reutilized. PMID- 11886771 TI - Immunopeptides in the defense reactions of Glossina morsitans to bacterial and Trypanosoma brucei brucei infections. AB - Several dipteran insects are vectors of parasites causing major human infectious diseases. Among these, the tsetse fly, Glossina spp., is responsible for the transmission of trypanosomes, the pathogens responsible for sleeping sickness in Africa. A better understanding of insect-parasite interactions will help establish new strategies to fight this important often fatal disease. Antimicrobial peptides (AMPs) are part of the humoral immune response in insects during bacterial, fungal and parasitic infections. Here, we studied the immune response of Glossina morsitans to bacteria and to Trypanosoma brucei brucei by analyzing the synthesis of AMPs as markers of the humoral immune response. By reversed-phase chromatography, mass spectrometry analysis, Edman degradation and in vitro antimicrobial assays of the hemolymph of immune-challenged adults of G. morsitans, we identified three AMPs: a cecropin, an attacin and a defensin. These three AMPs were found to be induced upon systemic bacterial infection and also after per os infections by bacteria and parasites. PMID- 11886772 TI - Fork head alternative binding drives stage-specific gene expression in the silk gland of Bombyx mori. AB - Here, we identified the main transactivator of fhx, the gene encoding the silk protein fibrohexamerin in posterior silk gland cells (PSG), as the homeotic SGF1/fork head factor. The same factor also stimulates sericin-1, another silk protein encoding gene, in the middle silk gland cells. SGF1/fork head is present in the silk gland nuclei during the whole course of larval life, but its binding to the fhx promoter occurs at intermolt and not during molt, when fhx is respectively turned on and off. The alternative binding of the factor is associated with specific changes in the fhx chromatin topology in PSG cells. Taken together, our results show that stabilization of SGF1/fork head to its target sequence is critical to promote fhx transcription at each intermolt. We also found that fhx is characterized by a PSG-specific DNase I hypersensitive site in the first intron, present during molt and intermolt, i.e. independent of the transcriptional status of the gene. All these data suggest that differential chromatin accessibility and fork head activation are crucial in controlling the spatial and temporal regulation of the fhx gene in the posterior silk gland cells. PMID- 11886773 TI - Molecular identification of an endosymbiotic bacterium associated with pederin biosynthesis in Paederus sabaeus (Coleoptera: Staphylinidae). AB - Biosynthesis of the structurally complex hemolymph toxin pederin is an eminent character of Paederus females. For that capability, however, they rely on endosymbiotic bacteria that are lacking in aposymbiotic females. The bacterial inhabitants of the two phenotypes in Paederus sabaeus are evaluated in a PCR based analysis of 16S rDNA. A certain fragment, which is not found in aposymbiotic females, is highly dominant in the other, biosynthesizing females and thus identifies the endosymbiont. Its DNA sequence reveals a member of the gamma subdivision of the Proteobacteria that is clustered within the genus Pseudomonas (sensu stricto) as it is most closely related to Pseudomonas aeruginosa. These bacteria appear as the hypothesized common producers of pederin and the pederin family of analogs from marine sponges. PMID- 11886774 TI - Synergistic interaction between two cockroach sodium channel mutations and a tobacco budworm sodium channel mutation in reducing channel sensitivity to a pyrethroid insecticide. AB - Pyrethroid insecticide resistance due to reduced nerve sensitivity, known as knockdown resistance (kdr or kdr-type), is linked to multiple point mutations in the para-homologous sodium channel genes. Previously we demonstrated that two mutations (E434K and C764R) in the German cockroach sodium channel greatly enhanced the ability of the L993F mutation (a known kdr -type mutation) to reduce sodium channel sensitivity to deltamethrin, a pyrethroid insecticide. Neither E434K nor C764R alone, however, altered sodium channel sensitivity. To examine whether E434K and C764R also enhance the effect of pyrethroid resistance associated sodium channel mutations identified in other insects, we introduced a V to M mutation (V409M) into the cockroach sodium channel protein at the position that corresponds to the V421M mutation in the Heliothis virescens sodium channel protein. We found that the V409M mutation alone modified the gating properties of the sodium channel and reduced channel sensitivity to deltamethrin by 10-fold. Combining the V409M mutation with either the E434K or C764K alone did not reduce the V409M channel sensitivity to deltamethrin further. However, the triple mutation combination (V409M, E434K and C764R) dramatically reduced channel sensitivity by 100-fold compared with the wild-type channel. These results suggest that the E434K and C764R mutations are important modifiers of sodium channel sensitivity to pyrethroid insecticides. PMID- 11886775 TI - Effects of a potato cysteine proteinase inhibitor on midgut proteolytic enzyme activity and growth of the southern corn rootworm, Diabrotica undecimpunctata howardi (Coleoptera: Chrysomelidae). AB - The major proteinase activity in extracts of larval midguts from the southern corn rootworm (SCR), Diabrotica undecimpunctata howardi, was identified as a cysteine proteinase that prefers substrates containing an arginine residue in the P1 position. Gelatin-zymogram analysis of the midgut proteinases indicated that the artificial diet-fed SCR, corn root-fed SCR, and root-fed western corn rootworms (Diabrotica virgifera virgifera) possess a single major proteinase with an apparent molecular mass of 25kDa and several minor proteinases. Similar proteinase activity pH profiles were exhibited by root-fed and diet-fed rootworms with the optimal activity being slightly acidic. Rootworm larvae reared on corn roots exhibited significantly less caseinolytic activity than those reared on the artificial diet. Midgut proteolytic activity from SCR was most sensitive to inhibition by inhibitors of cysteine proteinases. Furthermore, rootworm proteinase activity was particularly sensitive to inhibition by a commercial protein preparation from potato tubers (PIN-II). One of the proteins, potato cysteine proteinase inhibitor-10', PCPI-10', obtained from PIN-II by ion-exchange chromatography, was the major source of inhibitory activity against rootworm proteinase activity. PCPI-10' and E-64 were of comparable potency as inhibitors of southern corn rootworm proteinase activity (IC(50) =31 and 35nM, respectively) and substantially more effective than chicken egg white cystatin (IC(50) =121nM). Incorporation of PCPI-10' into the diet of SCR larvae in feeding trials resulted in a significant increase in mortality and growth inhibition. We suggest that expression of inhibitors such as PCPI-10' by transgenic corn plants in the field is a potentially attractive method of host plant resistance to these Diabrotica species. PMID- 11886776 TI - Mechanisms of resistance to organophosphates in Tetranychus urticae (Acari: Tetranychidae) from Greece. AB - We investigated the mechanisms conferring resistance to methyl-parathion (44 fold) and to methomyl (8-fold) in Tetranychus urticae from Greece by studying the effect of synergists on the resistance and the kinetic characteristics of various enzymes in a resistant strain (RLAB) and a susceptible reference strain (SAMB). It is shown that S,S,S-tributyl phosphorotrithioate, a synergist that inhibits esterases and glutathione S-transferases, and piperonyl butoxide, a synergist that inhibits cytochrome P450 mediated monooxygenases, did not affect the level of methyl-parathion or methomyl resistance in RLAB and that resistance ratios to both insecticides did not change significantly in the presence of either synergist. Isoelectric focusing of esterase allozymes on single mites revealed no differences in staining intensity and glutathione S-transferase activity was not significantly different in the two strains. The activity of two cytochrome P450 monooxygenase groups was compared. No significant difference of 7-ethoxyresorufin O-diethylase activity was observed between strains that were two-fold higher in RLAB than in SAMB. The kinetic characteristics of acetylcholinesterase, the target enzyme of organophosphates and carbamates, revealed that acetylcholinesterase in RLAB was less sensitive to inhibition by paraoxon and methomyl in comparison with SAMB. I(50), the inhibitor concentration inducing 50% decrease of acetylcholinesterase activity was greater (119- and 50-fold with paraoxon and methomyl, respectively) and the bimolecular constant k(i) was lower (39- and 47-fold with paraoxon and methomyl, respectively) in RLAB compared to SAMB. PMID- 11886777 TI - Expression and characterization of a novel class of glutathione S-transferase from Anopheles dirus. AB - A new Anopheles dirus glutathione S-transferase (GST) has been obtained and named adGST4-1. Both genomic DNA and cDNA for heterologous expression were acquired. The genomic sequence was 3188bp and consisted of the GST gene as well as flanking sequence. The flanking sequence was analyzed for possible regulatory elements that would control gene expression. In Drosophila several of these elements have been shown to be involved in development and cell differentiation. The deduced amino acid sequence has low identity compared with the four alternatively spliced enzymes, adGST1-1 to 1-4, from another An. dirus GST gene adgst1AS1. The percent identities are 30--40% and 11--12% comparing adGST4-1 to insect GSTs from Delta and Sigma classes, respectively. Enzyme characterization of adGST4-1 shows it to be distinct from the other An. dirus GSTs because of low enzyme activity for customary GST substrates including 1-chloro-2, 4-dinitrobenzene (CDNB). However, this enzyme has a greater affinity of interaction with pyrethroids compared to the other An. dirus GSTs. PMID- 11886778 TI - Prostaglandin biosynthesis by midgut tissue isolated from the tobacco hornworm, Manduca sexta. AB - We describe prostaglandin (PG) biosynthesis by isolated midgut preparations from tobacco hornworms, Manduca sexta. Microsomal-enriched midgut preparations yielded four PGs, PGA/B(2), PGD(2), PGE(2) and PGF(2alpha), all of which were confirmed by analysis on gas chromatography--mass spectrometry (GC--MS). PGA and PGB are double bond isomers which do not resolve on TLC but do resolve by GC; for convenience, we use the single term PGA(2) for this product. PGA(2) was the major product under most conditions. The midgut preparations were sensitive to reaction conditions, including radioactive substrate, protein concentration (optimal at 1mg/reaction), reaction time (optimal at 0.5 min), temperature (optimal at 22 degrees C), buffer pH (highest at pH 6), and the presence of a co-factor cocktail composed of reduced glutathione, hydroquinine and hemoglobin. In vitro PG biosynthesis was inhibited by two cyclooxygenase inhibitors, indomethacin and naproxen. Subcellular localization of PG biosynthetic activity in midgut preparations, determined by ultracentrifugation, revealed the presence of PG biosynthetic activity in the cytosolic and microsomal fractions, although most activity was found in the cytosolic fractions. This is similar to other invertebrates, and different from mammalian preparations, in which the activity is exclusively associated with the microsomal fractions. Midgut preparations from M. sexta pupae, adult cockroach, Periplaneta americana, and corn ear worms, Helicoverpa zea, also produced the same four major PG products. We infer that insect midguts are competent to biosynthesize PGs, and speculate they exert important, albeit unrevealed, actions in midgut physiology. PMID- 11886780 TI - Molecular cloning and partial characterization of a trypsin-like protein in salivary glands of Lygus hesperus (Hemiptera: Miridae). AB - Trypsin-like enzymes from the salivary gland complex (SGC) of Lygus hesperus Knight were partially purified by preparative isoelectric focusing (IEF). Enzyme active against Nalpha-benzoyl-L-arginine-p-nitroanilide (BApNA) focused at approximately pH 10 during IEF. This alkaline fraction gave a single activity band when analyzed with casein zymograms. The serine proteinase inhibitors, phenylmethylsulfonyl fluoride (PMSF) and lima bean trypsin inhibitor, completely inhibited or suppressed the caseinolytic activity in the crude salivary gland extract as well as the IEF-purified sample. Chicken egg white trypsin inhibitor also inhibited the IEF-purified sample but was not effective against a major caseinolytic band in the crude salivary gland extract. These data indicated the presence of serine proteinases in the SGC of L. hesperus. Cloning and sequencing of a trypsin-like precursor cDNA provided additional direct evidence for serine proteinases in L. hesperus. The encoded trypsin-like protein included amino acid sequence motifs, which are conserved with five homologous serine proteinases from other insects. Typical features of the putative trypsin-like protein from L. hesperus included residues in the serine proteinase active site (His(89), Asp(139), Ser(229)), conserved cysteine residues for disulfide bridges, residues (Asp(223), Gly(252), Gly(262)) that determine trypsin specificity, and both zymogen signal and activation peptides. PMID- 11886781 TI - Developmental profiles of ecdysteroids, ecdysteroid receptor mRNAs and DNA binding properties of ecdysteroid receptors in the Ixodid tick Amblyomma americanum (L.). AB - Total body ecdysteroid titers were determined at specific stages during the larval and nymphal life of Amblyomma americanum (L.). One ecdysteroid peak was observed following the completion of larval apolysis. However, two distinct ecdysteroid peaks occurred at a comparable stage in the nymphal molting cycle. The first occurred following apolysis and the second peak occurred at about the time of ecdysis. When whole body profiles of EcR and RXR mRNAs were examined during the molting cycle using RT-PCR, the expression of both AamEcR and AamRXR mRNAs was shown to be correlated with the ecdysteroid titer. Using an electrophoretic gel mobility shift assay, it was demonstrated that AamEcR*AamRXR1, but not AamEcR*AamRXR2, exhibits broad DNA binding specificity, forming complexes with a variety of synthetic direct repeat and palindromic nuclear response elements with the half-site consensus AGGTCA. These data suggest that functional differences may exist between the AamRXR1 and AamRXR2 proteins. PMID- 11886779 TI - Novel sodium channel gene mutations in Blattella germanica reduce the sensitivity of expressed channels to deltamethrin. AB - Pyrethroid insecticides alter the normal gating of voltage-gated sodium channels in the nervous system. Three sodium channel mutations (E434K, C764R, L993F) were recently identified in pyrethroid resistant German cockroach populations. In this report, we show that the L993F mutation decreased sodium channel sensitivity to the pyrethroid, deltamethrin, by five-fold in Xenopus oocytes. In contrast, neither E434K nor C764R alone decreased channel sensitivity to deltamethrin. However, E434K or C764R combined with L993F reduced deltamethrin sensitivity by 100-fold. Furthermore, concomitant presence of all three mutations (KRF) reduced channel sensitivity to deltamethrin by 500-fold. None of the mutations significantly affected channel gating. However, sodium current amplitudes from the mutant sodium channel carrying either E434K or C764R alone were much reduced compared to those of the wild-type channel or the channel carrying the double or triple mutations (KF, RF and KRF). These results indicated that evolution of sodium channel insensitivity in the German cockroach is achieved by sequential selection of a primary mutation L993F and two secondary mutations E434K and C764R, and concomitant presence of all three mutations dramatically reduced sodium channel sensitivity to deltamethrin. PMID- 11886782 TI - cDNA cloning of calcineurin heterosubunits from the pheromone gland of the silkmoth, Bombyx mori. AB - Pheromone biosynthesis activating neuropeptide (PBAN) stimulates the step of fatty acyl reduction in the pheromone biosynthetic pathway of the silkmoth, Bombyx mori. It has been suggested that the intracellular signal transduction of PBAN in B. mori involves Ca(2+), calmodulin, and calcineurin (also known as protein phosphatase 2B). We have cloned two cDNAs encoding calcineurin heterosubunits from a pheromone gland cDNA library of B. mori. The 2,996-bp clone predicts a 495-amino acid protein homologous to the catalytic subunit calcineurin A (CnA) with a molecular mass of 55,968. The deduced amino acid sequence well conserves the calcineurin B (CnB)-binding domain and two subdomains, a calmodulin binding and an autoinhibitory domain, showing 77-85% and 82% identities to the isoforms of Drosophila melanogaster CnA and human CnA, respectively. On the other hand, the 820-bp clone predicts a 170-amino acid protein homologous to the regulatory subunit CnB with a molecular mass of 19,357. The deduced amino acid sequence well conserves four EF-hand type calcium-binding structures, showing 95% and about 85% identities to D. melanogaster CnB and mammalian CnBs, respectively. A yeast two-hybrid system has demonstrated the molecular interaction between B. mori CnA and CnB. Northern blot analyses revealed that both CnA and CnB genes were expressed in various larval and adult tissues of B. mori. Both transcripts detected in the pheromone gland three days before adult eclosion increased by the day before eclosion and the mRNA levels were found to be high even two days after adult eclosion. Immunohistochemical analysis has revealed that B. mori calcineurin is localized in the cytoplasm of the pheromone-producing cells. PMID- 11886784 TI - An analysis of the binding of cocaine analogues to the monoamine transporters using tensor decomposition 3-d QSAR. AB - The conformation and alignment of cocaine analogues bound to the monoamine transporter proteins were explored using the tensor decomposition 3-D QSAR method. It is proposed from these calculations that the bound conformation of these ligands to the three transporter proteins has the 3beta-aryl substituent in a conformation in which the aryl group is orthogonal or approximately orthogonal to the tropane ring. Based on these results, rigid and semi-rigid tropane analogues were designed, synthesized and their affinities for the monoamine transporters were determined. PMID- 11886785 TI - Coupling of isoprenoid triflates with organoboron nucleophiles: synthesis and biological evaluation of geranylgeranyl diphosphate analogues. AB - The Suzuki coupling reaction has been used to introduce a methyl group derived from commercially available methylboronic acid into a vinyl triflate. This has led to a concise synthesis of all-trans-geranylgeraniol, with the key step being the palladium-catalyzed, silver-mediated methylation of triflate to give ethyl geranylgeranoate. This coupling protocol has also been used to produce the novel geranylgeranyl diphosphate (GGPP) analogue 3-phenyl-3-desmethylgeranylgeranyl diphosphate (3-PhGGPP, ). Our previously developed organocuprate coupling protocol has been used to introduce the cyclopropyl and tert-butyl moieties into the 3-position of vinyl triflate. The four GGPP analogues 3-vinyl-3 desmethylgeranylgeranyl diphosphate (3-vGGPP, ), 3-cyclopropyl-3 desmethylgeranylgeranyl diphosphate (3-cpGGPP, ), 3-tert-butyl-3-desmethyl geranylgeranyl diphosphate (3-tbGGPP, ), and were then evaluated as potential inhibitors of recombinant yeast protein-geranylgeranyl transferase I (PGGTase I). The potential mechanism-based inhibitors 3-vGGPP and 3-cpGGPP did not exhibit time-dependent inactivation of PGGTase I. Instead, both analogues were alternative substrates, in accord with the interaction of the corresponding farnesyl analogues 3-vFPP and 3-cpFPP with PFTase. The tert-butyl and phenyl analogues were not substrates, but were instead competitive inhibitors of PGGTase I. Note that all four of the GGPP analogues were bound less tightly by the enzyme than the natural substrate, in contrast to the behavior of the 3-substituted FPP analogues. PMID- 11886786 TI - Acetamidoquinone and acetamidohydroxy derivatives as inhibitors for both dihydroxyacetamido epoxidase and dehydrogenase. AB - A series of monohydroxy and dihydroxyacetanilides, acetamidoquinones and bromoacetamidoquinones have been synthesised and tested as substrates and/or inhibitors of highly purified dihydroxyacetamido epoxidase (DHAE) and dihydroxy acetamido dehydrogenase (DHADH) from Streptomyces LL-C10337. None was found to act as substrates but many selectively inhibit the enzymes. Kinetic analysis has shown that all the compounds act as reversible competitive inhibitors with respect to the substrates 2,5-dihydroxyacetanilide and 2,3-epoxy-1,4-benzoquinone 5-acetanilide. Monohydroxy acetanilides showed weak inhibition to these enzymes compared to the dihydroxy derivatives while the more powerful inhibitors were the benzoquinoneacetanilide and its 5-bromo equivalent. PMID- 11886787 TI - 5'-alkyl-benzothiadiazides: a new subgroup of AMPA receptor modulators with improved affinity. AB - AMPA receptors form a major subdivision of the glutamate receptor family that mediates excitatory synaptic transmission in the brain. Currents through AMPA receptors can be up- or down-regulated by compounds that allosterically modulate receptor kinetics through binding sites distinct from that for glutamate. One of those modulators is the benzothiadiazide IDRA-21 which has been reported to enhance synaptic transmission and be effective in behavioral tests, but typically requires threshold concentrations of at least 100 microM to be active in vitro. In this study, new benzothiadiazides were developed with IDRA-21 as lead compound and examined for their potency in modulating AMPA receptor kinetics. A significant increase in drug affinity was obtained by alkyl substitution at the 5'-position of IDRA-21; substitutions at other positions of the benzothiadiazide core generally did not yield a further gain in affinity and in some cases abolished drug binding. The 5'-ethyl derivative exhibited an EC(50) value in the order of 22 microM which represents about a 30-fold gain in affinity over that of IDRA-21. The EC(50) value is comparable to that of cyclothiazide, the most potent benzothiadiazide drug, but the effects on AMPA receptors differed substantially between these two compounds in that the 5'-ethyl derivative of IDRA-21 greatly increased the binding affinity for receptor agonists whereas cyclothiazide is known to reduce agonist binding. The structure--activity relationships reported here thus offer to provide new insights how receptor kinetics is linked to particular aspects of receptor--drug interactions. PMID- 11886788 TI - Polyene substrates with unusual methylation patterns to probe the active sites of three catalytic antibodies. AB - The synthesis of two tetraenes that differ in their methylation pattern from the natural substrate in lanosterol biosynthesis, 2,3-oxidosqualene, and their examination with three catalytic antibodies is described. The design of these novel, linear terpenoid structures was governed by initial results obtained from the characterization of the three catalytic antibodies. These were generated by immunization with a steroidal hapten that mimics multicyclization without the necessity for anti-Markovnikov additions or ring expansions. Such a reaction cascade would represent a more 'primitive' version compared to the oxidosqualene cyclization observed in lanosterol, cycloartenol and beta-amyrin biosynthesis and would not require a tail-to-tail connection of the third and fourth isoprene unit as seen in squalene. The first tetraene design (A) only contains trisubstituted double bonds and hence its synthesis starts from farnesol and tris-norgeraniol. The second tetraene design (B) is considered the more precise match to the inducing hapten that generated the antibody collections by exhibiting one disubstituted double bond and its synthesis utilizes a tris-norgeraniol derivative and a symmetrical bis-allylic alcohol as key building blocks. Chromatographic comparison studies lead to the conclusion that the currently studied antibodies also produce monocyclic products from the two substrates as has been formerly observed with a squalene-derived substrate. In contrast, 2,3 oxidosqualene is not accepted by these catalysts supporting the notion that the current substrates are fully bound by recognition of both terminal functional groups. PMID- 11886789 TI - Synthesis and biological properties of amino acid amide ligand-based pyridinioalkanoyl thioesters as anti-HIV agents. AB - Hyper-mutable retroviruses such as HIV can become rapidly resistant to drugs used to treat infection. Strategies for coping with drug-resistant strains of virus include combination therapies, using viral protease and reverse transcriptase inhibitors. Another approach is the development of antiviral agents that attack mutationally nonpermissive targets that have functions essential for viral replication. Thus, the highly conserved nucleocapsid protein, NCp7, was chosen as a prime target in our search for novel anti-HIV agents that can overcome the problem of viral drug resistance. Recently, we reported (J. Med. Chem. 1999, 42, 67) a novel chemotype, the pyridinioalkanoyl thioesters (PATEs), based on 2 mercaptobenzamides as the thiol component and having its amide nitrogen substituted with various phenylsulfonyl moieties. These compounds were identified as relatively nontoxic anti-HIV agents in the XTT cytoprotection assay. In this study, we wish to report a separate genre of active PATEs wherein the thiol component consists of an N-2-mercaptobenzoyl-amino acid derivative. Active derivatives (EC(50) < 10 microM) reported herein were confined to amino acid primary amides or methyl amides having side chains no larger than isobutyl. Amino acids terminating in free carboxyl or carboxylic acid ester groups were mostly inactive. Selected compounds were shown to be active on chronically infected CEM/SK-1, TNFalpha-induced U1, ACH-2 cells and virucidal on cell-free virus, latently infected U1 cells and acutely infected primary peripheral blood mononuclear cells (PBMCs). PMID- 11886791 TI - Novel irreversible butyrylcholinesterase inhibitors: 2-chloro-1-(substituted phenyl)ethylphosphonic acids. AB - 2-Chloroethylphosphonic acid (ethephon) as the dianion phosphorylates butyrylcholinesterase (BChE) at its active site. In contrast, the classical organophosphorus esterase inhibitors include substituted-phenyl dialkylphosphates (e.g., paraoxon) with electron-withdrawing aryl substituents. The chloroethyl and substituted-phenyl moieties are combined in this study as 2-chloro-1-(substituted phenyl)ethylphosphonic acids (1) to define the structure--activity relationships and mechanism of BChE inhibition by ethephon and its analogues. Phenyl substituents considered are 3- and 4-nitro, 3- and 4-dimethylamino, and 3- and 4 trimethylammonium. Phosphonic acids were synthesized via the corresponding O,O diethyl phosphonate precursors followed by deprotection with trimethylsilyl bromide. They decompose under basic conditions about 100-fold faster than ethephon to yield the corresponding styrene derivatives. Electron-withdrawing substituents on the phenyl ring decrease the hydrolysis rate while electron donating substituents increase the rate. The 4-trimethylammonium analogue has the highest affinity (K(i)=180 microM) and potency (IC(50)=19 microM) in first binding reversibly at the substrate site (possibly with stabilization in a dianion--monoanion environment) and then progressively and irreversibly inhibiting the enzyme activity. These observations suggest dissociation of chloride as the first and rate-limiting step both in the hydrolysis and by analogy in phosphorylation of BChE by bound at the active site. PMID- 11886790 TI - A convenient synthesis and hepatoprotective activity of imidazo[1,2 c]pyrimido[5,4-e]pyrimidine, tetraazaacenaphthene and tetraazaphenalene from cyclic ketene aminals through tandem addition-cyclization reactions. AB - A novel one-pot synthesis of imidazo[1,2-c]pyrimido[5,4-e]pyrimidinones (2), tetraazaacenaphthene-3,6-diones (4), tetarazaphenalene-1,7-dione (4d) is delineated from the reaction of cyclic ketene aminal (1) and alkyl or aryl isothiocyanate through tandem addition-cyclization reactions. However, reaction of ketene aminal (1a) with alkyl isothiocyanate only yielded angularly cyclized product 5 which did not react further to yield 6. The structure of 2c and 4d was ascertained by single crystal X-ray diffraction analysis which demonstrated a network of various inter- and intramolecular interactions, responsible for the stability and packing of the molecules in the crystalline state. Some of the compounds (2a--h) were screened for hepatoprotective activity but only 2a was found most effective. PMID- 11886792 TI - Structure--activity relationships among novel phenoxybenzamine-related beta chloroethylamines. AB - A series of beta-chloroethylamines 5--18, structurally related to the irreversible alpha(1)-adrenoceptor antagonist phenoxybenzamine [PB, N-benzyl-N-(2 chloroethyl)-N-(1-methyl-2-phenoxyethyl)amine hydrochloride, 1] and the competitive antagonist WB4101 [N-(2,3-dihydro-1,4-benzodioxin-2-ylmethyl)-N-[2 (2,6-dimethoxyphenoxy)ethyl]amine hydrochloride, 2], were synthesized and evaluated for their activity at alpha-adrenoceptors of the epididymal and the prostatic portion of young CD rat vas deferens. All compounds displayed irreversible antagonist activity. Most of them showed similar antagonism at both alpha(1)- and alpha(2)-adrenoceptors, whereas compounds 13 and 18, lacking substituents on both the phenoxy group and the oxyamino carbon chain, displayed a moderate alpha(1)-adrenoceptor selectivity (10--35 times), which was comparable to that of PB. Compounds 14 and 15, belonging to the benzyl series and bearing, respectively, a 2-ethoxyphenoxy and a 2-i-propoxyphenoxy moiety, were the most potent alpha(1)-adrenoceptor antagonists with an affinity value similar to that of PB (pIC(50) values of 7.17 and 7.06 versus 7.27). Interestingly, several compounds were able to distinguish two alpha(1)-adrenoceptor subtypes in the epididymal tissue, as revealed by the discontinuity of their inhibition curves. A mean ratio of 24:76 for these alpha(1)-adrenoceptors was determined from compounds 8--10, 12, and 15--17. Furthermore, compounds 9, 10, 12, 16a, and 16b showed higher affinity towards the minor population of receptors, whereas compounds 8, 15, and 17 preferentially inhibited the major population of alpha(1) adrenoceptors. In addition, selected pharmacological experiments demonstrated the complementary antagonism of the two series of compounds and their different, preferential affinity for one of the two alpha(1)-adrenoceptor subtypes. In conclusion, we found beta-chloroethylamines that demonstrate a multiplicity of alpha(1)-adrenoceptors in the epididymal portion of young CD rat vas deferens and, as a consequence, they are possible useful tools for alpha(1)-adrenoceptor characterization. PMID- 11886793 TI - Fatty acid esters of juvenoid alcohols as insect hormonogen agents (juvenogens). AB - A series of 8 new juvenogens (3--10) was prepared starting from a pair of isomeric insect juvenile hormone bioanalogues ( and ). The biological activity of the juvenogens -- was tested for their effect on reproduction of the blowfly Neobellieria (Sarcophaga) bullata and for the juvenilizing activity on the termite Prorhinotermes simplex. Results of biological screening are important in structure--activity studies and promising for potential practical application of some of the juvenogens studied, especially against termites. PMID- 11886794 TI - The 1.76 A resolution crystal structure of glycogen phosphorylase B complexed with glucose, and CP320626, a potential antidiabetic drug. AB - CP320626, a potential antidiabetic drug, inhibits glycogen phosphorylase in synergism with glucose. To elucidate the structural basis of synergistic inhibition, we determined the structure of muscle glycogen phosphorylase b (MGPb) complexed with both glucose and CP320626 at 1.76 A resolution, and refined to a crystallographic R value of 0.211 (R(free)=0.235). CP320626 binds at a novel allosteric site, which is some 33 A from the catalytic site, where glucose binds. The high resolution structure allows unambiguous definition of the conformation of the 1-acetyl-4-hydroxy-piperidine ring supported by theoretical energy calculations. Both CP320626 and glucose promote the less active T-state, thereby explaining their synergistic inhibition. Structural comparison of MGPb--glucose- CP320626 complex with liver glycogen phosphorylase a (LGPa) complexed with a related compound (CP403700) show that the ligand binding site is conserved in LGPa. PMID- 11886795 TI - A novel approach towards studying non-genotoxic enediynes as potential anticancer therapeutics. AB - A novel uracil-containing enediyne was synthesized by the fusion at N(1) and N(3) of uracil with an 11-membered cyclic enediyne. Compound was found to be stable against cycloaromatization at 80 degreesC. Thus, it did not cause DNA-damage. Unlike other alkylated uracil derivatives 2--6, highly strained uracil-containing enediyne was reacted with methyl thioglycolate at 25 degreesC to produce uracil () and linear enediyne. This reactivity toward a sulfhydryl group may play a significant role in the mechanism by which compound directed its cytotoxicity toward tumor cell lines. Tumor cells were found to be more susceptible to enediyne than normal human embryonic lung cells. A combination of with adriamycin or 1-(beta-D-arabinofuranosyl)cytosine resulted in synergistic anticancer activity against murine L1210 and P388 leukemias, Sarcoma 180, and human CCRF- CEM lymphoblastic leukemia. After treatment of Molt-4 cells with uracil containing enediyne, light microscope examination demonstrated the presence of cell shrinkage and nuclear segmentation. Treatment of cultured Molt-4 human leukemia cells with enediyne resulted in a time-dependent depletion of glutathione (GSH) whereas the exposure of the cells to the GSH precursor N acetylcysteine (NAC) resulted in a substantial suppression of this effect. As such, involvement of GSH depletion in the process of apoptosis may explain the mechanism of action of non-genotoxic enediyne against malignant tumor cell lines. PMID- 11886796 TI - Recognition of bulged DNA by a neocarzinostatin product via an induced fit mechanism. AB - The binding of the wedge-shaped isostructural analogue of the biradical species of the chromphore of antitumor antibiotic neocarzinostatin to sequence-specific bulged DNAs results in alterations in ellipticity of the DNAs. Circular dichroism (CD) spectroscopic results suggest that the drug specifically recognizes bulges of DNA via a combination of conformational selection and induced fit, not by binding to a preorganized site. Analysis of circular dichroism spectra indicates that the degree of induced fit observed is primarily a consequence of optimising van der Waals contacts with the walls of the bulge cavity. The effective recognition of the bulge site on duplex DNA appears to depend to a significant extent on the bent groove space being flexible enough to be able to adopt the geometrically optimal conformation compatible with the wedge-shaped drug molecule, rather than involving 'lock and key' recognition. The spectroscopic results indicate a change of DNA conformation, consistent with an allosteric binding model. Spectroscopic studies with various bulged DNAs also reveal that the binding strength directly correlates with the stability of the bulge structures. PMID- 11886797 TI - Substituted indoloquinolines as new antifungal agents. AB - Cryptolepine (2) possesses desirable properties to serve as a lead in developing new antifungal agents. Using SAR techniques, several analogues of cryptolepine were designed to increase potency and to broaden the antifungal spectrum over several opportunistic microorganisms. A number of 2-substituted indoloquinolines have been synthesized and evaluated in antifungal screens and several have been shown to increase potency and expand the antifungal spectrum of cryptolepine. Comparison of MICs of a number of these analogues with standard antifungal agents, shows them to be comparable to Amphotericin B and Ketoconazole. PMID- 11886798 TI - Discovery of diaminobutane derivatives as Ca(2+)-permeable AMPA receptor antagonists. AB - We designed and synthesized a series of the polyamine derivatives as potent Ca(2+)-permeable AMPA receptor antagonists. In the course of this study, we found that the polyamine derivatives exhibited strong hypotensive activity which was undesirable activity for neuroprotective agents. Therefore, we tried to find non hypotensive antagonists by structural modification of such compounds. Through this derivatization, we obtained the diamine compounds having desired profiles. Especially, compound 8f, which was non-hypotensive and potent Ca(2+)-permeable AMPA receptor antagonist, showed neuroprotective effects in transient global ischemia models in gerbils. PMID- 11886799 TI - QSAR studies on antimalarial substituted phenyl analogues and their N(omega) oxides. AB - A quantitative structure--activity relationship (QSAR) study on a series of substituted phenyl analogues 5-[(7-chloro-4-quinolinyl)amino]-3 [(alkylamino)methyl](1,1'-biphenyl)-2-ols and their N(omega)-oxides was made using various combinations of electronic and topological parameters. Several statistically significant regression expressions were obtained using multiple regression analyses. These regressions may be considered as mathematical models for investigating antimalarial activities of the compounds under present study. The antimalarial activity mechanism was investigated using combinations of E(L) and E(H), independently with other molecular descriptors. PMID- 11886800 TI - 3-D-QSAR analysis of N-(3-acyloxy-2-benzylpropyl)-N' dihydroxytetrahydrobenzazepine and tetrahydroisoquinoline and N-(3-acyloxy-2 benzylpropyl)-N'-(4-hydroxy-3-methoxybenzyl) thioureas analogues as potent vanilloid receptor ligands. AB - 3-D-Quantitative structure--activity relationships of N-(3-acyloxy-2 benzylpropyl)-N'-dihydroxytetrahydro-benzazepine and tetrahydroisoquinoline and N (3-acyloxy-2-benzylpropyl)-N'-(4-hydroxy-3-methoxybenzyl) thiourea analogues as potent vanilloid receptor ligands were investigated using the CoMFA and the COMSIA methods. The best CoMFA model obtained in this study from 29 substituted thiourea analogues is a two-component model with the following statistics. R(2)((cv))=0.407 and RMSE((cv))=0.532 for the cross-validation, and R(2)=0.705 and RMSE=0.375 for the fitted. The best COMSIA model obtained from the same 29 compounds is a two-component model with the following statistics: R(2)((cv))=0.336 and RMSE((cv))=0.563 for the cross-validation, and R(2)=0.693 and RMSE=0.382 for the fitted. PMID- 11886801 TI - Molecular orbital calculation for the model compounds of kainoid amino acids, agonists of excitatory amino acid receptors. Does the kainoid C4-substituent directly interact with the receptors? AB - Kainoid amino acids are agonists of the AMPA/kainate receptors and exhibit highly potent neuroexcitatory activity. From the results of extensive structure- activity relationship studies, we previously postulated that the C4-substituent of the kainoid amino acids interacts with an allosteric site of the glutamate receptor with electron-donating character. In order to investigate the mode of action in more detail, molecular orbital calculation for model compounds of the kainoid were performed. The results indicated that the HOMO energy level of the C4-substituent is involved in the potent neuroexcitatory activity, thus supporting our hypothesis. PMID- 11886802 TI - Aryl cyclopentadienyl tricarbonyl rhenium complexes: novel ligands for the estrogen receptor with potential use as estrogen radiopharmaceuticals. AB - The need for imaging agents for estrogen receptor positive (ER+) tumors that are both cost effective and widely available, as well as the need for novel radiotherapeutic agents for the treatment of breast cancer, has prompted us to investigate cyclopentadienyl tricarbonyl metal [CpMet(CO)(3), Met=Re, Tc-99m] complexes that bind well to the ER. Thus, we have prepared a series of p hydroxyphenyl-substituted CpRe(CO)(3) complexes and evaluated them (and, in some cases, their cyclopentadiene precursors) for binding to ER. These compounds constitute a new class of structurally integrated organometallic ligands for ER in which the CpMet(CO)(3 )organometallic unit forms the very structural core of these molecules and thus is necessarily intimately involved in their interaction with the receptor. The CpRe(CO)(3) compounds were prepared by reaction of the lithium salt of the arene-substituted cyclopentadiene with a suitable Re(CO)(3)(+) precursor, followed by deprotection of the methyl ether. The X-ray crystal structure of one of these analogues shows that it has the classical 'piano stool'-like geometry, with the alkyl groups directed upward, away from the tripodyl metal carbonyl base. The aryl-substituted CpRe(CO)(3) complexes that we have prepared all bind to the ER, some with affinity as great as 20% that of the native ligand, estradiol. In general, at least two p-hydroxyphenyl substituents and one to two alkyl groups attached to the organometallic cyclopentadienyl core are needed for high ER affinity. Where we have been able to make comparisons, the metal complexes bind to ER with an affinity greater than their cyclopentadiene precursors. The high affinity of some of these complexes indicates that the bulky Re(CO)(3) unit is able to exploit the considerable volume in the center of the ER ligand binding pocket that is not occupied by most ligands, a consideration that is supported by molecular modeling. The preparation of the best of these agents in technetium-99m labeled form is currently being investigated. PMID- 11886803 TI - Synthesis and biological activity of 1-phenylsulfonyl-4 phenylsulfonylaminopyrrolidine derivatives as thromboxane a(2) receptor antagonists. AB - The synthesis and biological activity of novel 1-phenylsulfonyl-4- phenylsulfonylaminopyrrolidine analogues are described. All compounds were produced through modification of the substituent formally corresponding to the 1,3-dioxane ring system and the omega-octenol side chain of thromboxane A(2) (TXA(2)), in reference to the structure of Daltroban. Several compounds were found to be potent TXA(2) receptor antagonists. Compound 51a was the most effective inhibitor of 9,11-epoxymethano PGH(2) (U-46619)-induced rat aortic strip contraction (IC(50)=0.48 nM). PMID- 11886804 TI - Certification of the critical importance of L-3-(2-naphthyl)alanine at position 3 of a specific CXCR4 inhibitor, T140, leads to an exploratory performance of its downsizing study. AB - We have previously found that a 14-amino acid residue-peptide, T140, inhibits infection of target cells by T cell line-tropic HIV-1 (X4-HIV-1) through its specific binding to a chemokine receptor, CXCR4. Here, the importance of an L-3 (2-naphthyl)alanine (Nal) residue at position 3 in T140 for high anti-HIV activity and inhibitory activity against Ca(2+) mobilization induced by stromal cell-derived factor (SDF)-1alpha-stimulation through CXCR4 has initially been shown by the synthesis and biological evaluation of several analogues, where Nal(3) is substituted by diverse aromatic amino acids. Next, the order of the N terminal 3 residues (Arg(1)-Arg(2)-Nal(3)) has been proved to be important from the structure--activity relationship (SAR) study shuffling these residues. Based on these results, we have found 10-residue peptides possessing modest anti-HIV activity by systematic antiviral evaluation of a series of synthetic, shortened analogues of T140. PMID- 11886805 TI - Binding of 1-benzopyran-4-one derivatives to aldose reductase: a free energy perturbation study. AB - The relative binding affinities to human aldose reductase (ALR2) of three new 7 hydroxy-2-benzyl-4H-1-benzopyran-4-one inhibitors were predicted by free energy perturbation (FEP) simulations. Molecular substitutions were specifically designed to investigate the role of hydrogen bonding at the active site of ALR2. Starting from the lead inhibitor 7-hydroxy-2-(4'-hydroxybenzyl)-4H-1-benzopyran-4 one, the 4'-hydroxyl was mutated to methyl and to trifluoromethyl, and an hydroxyl at position 8 was additionally introduced. Once synthesized and tested as inhibitors of ALR2, the compounds displayed variations of K(i) that were in qualitative to quantitative agreement with the calculated relative free energies of binding. The results, discussed in terms of balance between free energies of solvation and free energies of binding to ALR2, elucidate the importance of hydrogen bonding with Thr113 and with Trp111 and cofactor, and provide a rationale to the observed differences in binding affinities. PMID- 11886806 TI - Discovery of new inhibitors of aldose reductase from molecular docking and database screening. AB - Aldose reductase (ALR2) is a target enzyme for the treatment of diabetic complications. Owing to the limited number of currently available drugs for the treatment of diabetic complications, the discovery of new inhibitors of ALR2 that can potentially be optimized as drugs appears highly desirable. In this study, a molecular docking analysis of the structures of more than 127,000 organic compounds contained in the National Cancer Institute database was performed to find and score molecules that are complementary to ALR2. Besides retrieving several carboxylic acid derivatives, which are known to generally inhibit aldose reductase, docking proposed other families of putative inhibitors such as sulfonic acids, nitro-derivatives, sulfonamides and carbonyl derivatives. Twenty five compounds, chosen as the highest-scoring representatives of each of these families, were tested as aldose reductase inhibitors. Five of them were found to inhibit aldose reductase in the micromolar range. For these active compounds, selectivity with respect to the closely-related aldehyde reductase was determined by measuring the corresponding inhibitory activities. The structures of the complexes between the new lead inhibitors and aldose reductase, here refined with molecular mechanics and molecular dynamics calculations, suggest that new pharmacophoric groups can bind aldose reductase very efficiently. In the case of the family of the nitro-derivative inhibitors, a class of particularly interesting compounds, a round of optimizations was performed with the synthesis and biological evaluation of a series of derivatives aimed at testing the proposed binding mode and at improving interaction with active site residues. Starting from a hit compound having an IC(50) of 42 microM, the most potent compound synthesized showed a 10-fold increase in inhibitory activity and 10-fold selectivity with respect to ALR1, and structure--activity relationships of the designed compounds were in agreement with the proposed mode of binding at the active site. PMID- 11886807 TI - Synthesis and 31P NMR characterization of new low toxic highly sensitive pH probes designed for in vivo acidic pH studies. AB - With the aim to provide sensitive 31P NMR probes of intra- and extracellular pH gradients that may reach cellular acidic compartments in biological systems, new alpha-aminophosphonates were designed to meet basic requirements such as a low pK(a)s and a great chemical difference (Deltadelta(ab)) between the limiting 31P NMR chemical shifts in acidic (delta(a)) and basic (delta(b)) media. A series of six phosphorylated pyrrolidines and linear aminophosphonates were synthesized using aminophosphorylation reactions and were screened for cytotoxicity on cultured Muller cells. Among the compounds not being toxic under these conditions, three molecules were selected since they displayed the best in vitro (in several phosphate buffers and in a cytosol-like solution) properties as 31P NMR acidic pH markers, that is 3, 5 and 9, having the pK(a) values of 3.63, 5.89 and 5.66, respectively. The Deltadelta(ab) values of these pH markers were at least 3 times larger than that of standard 31P NMR probes, with a low sensitivity to ionic strength changes. From these data, it was proposed that 3, 5 and 9 could be used as reporting probes of subtle proton movements in acidic compartments, an area that still remains poorly investigated using non invasive 31P NMR methods. PMID- 11886808 TI - Novel anthracycline oligosaccharides: influence of chemical modifications of the carbohydrate moiety on biological activity. AB - Several observations highlight the importance of the carbohydrate moiety for the biological activity of antitumoural anthracyclines. Here is reported the synthesis, cytotoxicity and topoisomerase II-mediated DNA cleavage intensity of the new oligosaccharide anthracyclines 1--4 modified in the sugar residue. Evaluation of cytotoxic potency on different cell lines, resulted in quite similar values among the different analogues. On the other hand, topoisomerase II mediated DNA breaks level was different for the various compounds, and was not related to cytotoxicity, thus supporting previous observations reported for some monosaccharide anthracyclines modified in the carbohydrate portion. PMID- 11886809 TI - Resolution of (RS)-proglumide using lipase from Candida cylindraceae. AB - Proglumide is used in the treatment of neuropathic pain. It acts by inhibiting peptide cholecystokinin (CCK). Neural injury produces an elevation in plasma CCK. Proglumide has been also shown to augment the analgesic effect of sustained release morphine in neuropathic pain. Currently proglumide is administered as a racemic mixture. In the present study, an attempt is made to separate the racemic mixture of the drug using lipase obtained from Candida cylindracea by stereoselective esterification. Enzymatic stereoselective esterification was carried out in organic solvents. The resolution was studied using a chromatographic column with a chiral support and mass spectrometry. The reaction conditions for stereoselective esterification including amount of substrate, amount of enzyme, alcohol, solvent and temperature were optimised during the present investigation. Butanol and hexanol were found to be suitable for formation of S and R esters, respectively. Hexane was the best solvent for esterification and the optimum temperature was found to be 30 degreesC. PMID- 11886810 TI - Experimental and calculated shift in pK(a) upon binding of phosphotyrosine peptide to the SH2 domain of p56(lck). AB - The pH dependence of the affinity of a 11-mer phosphotyrosine (pY) peptide (EPQpYEEIPIYL-NH2) for the SH2 domain of the tyrosine kinase p56(lck) was investigated with surface plasmon resonance (SPR). From SPR competition experiments the affinity in solution was obtained. The pH dependence of the affinity in solution can be well described by a proton linkage model with a single pK(a) shift upon binding, from 6.1 to 4.7. This shift is ascribed to the transition from the -2 to the -1 ionisation state of the tyrosine phosphate group. Based on the X-ray structure for the complex with Lck SH2, a pK(a) value of 5.3 for the bound pY peptide was computed, modelling the solvated protein as a system of point charges in a continuum. With the phosphate in the -2 state the binding energy is 1.8 kcal/mol more favourable than for the -1 state, corresponding to a 20-fold higher affinity. A proper charge is relevant in the design of potential therapeutic Lck SH2 ligands with mimics for the metabolically unstable tyrosine phosphate group. PMID- 11886811 TI - QSAR of HIV-1 integrase inhibitors by genetic function approximation method. AB - Quantitative structure--activity relationship (QSAR) paradigm, using genetic function approximation (GFA) technique was used to examine the correlations between the calculated physicochemical descriptors and the in vitro activities (3'-processing and 3'-strand transfer inhibition) of a series of human immunodeficiency virus type 1 (HIV-1) integrase inhibitors. Depending on the chemical structure, all molecules were divided into two classes---catechols and noncatechols. Eighty-one molecules were used in the present study and they were divided into training set and test set. The training set in each class consisted of 35 molecules and QSAR models were generated separately for both catechols and noncatechols. Equations were evaluated using internal as well as external test set predictions. Models generated for catechols show that electronic, shape related, and thermodynamic parameters are important whereas for noncatechols, spatial, structural, and thermodynamic properties play an important role for the activity. PMID- 11886812 TI - Synthesis and biological evaluation of novel thioapio dideoxynucleosides. AB - On the basis of the bioisosteric rationale to apio dideoxynucleosides, novel thioapio dideoxynucleosides have been synthesized, starting from 1,3 dihydroxyacetone via thioapio sugar acetate as a key intermediate. The intermediate was condensed with silylated pyrimidine bases such as N(4) benzoylcytosine, uracil or thymine in the presence of TMSOTf to give the beta anomers and alpha-anomers, respectively. The intermediate was also condensed with silylated 6-chloropurine to give the 6-chloropurine derivatives and which were converted to adenine derivatives and, N(6)-methyladenine derivatives and, and hypoxanthine derivatives and, respectively. The guanine analogues and were also synthesized from the condensation of sugar acetate with 2-acetamido-6 chloropurine. All synthesized final compounds were tested against HIV-1. Most of the synthesized compounds exhibited toxicity-dependent anti-HIV-1 activity, among which 6-chloropurine derivative was found to be the most cytotoxic and showed good cytotoxicity against colon cancer cell lines. Although we could not find good anti-HIV agents in this study, findings of some anticancer activity in this series will allow this class of nucleosides to be the new template for the development of new anticancer agents (Fig. 1). PMID- 11886813 TI - The discovery of YM-60828: a potent, selective and orally-bioavailable factor Xa inhibitor. AB - Since Factor Xa (FXa) is well known to play a central role in thrombosis and hemostasis, inhibition of FXa is an attractive target for antithrombotic strategies. As a part of our investigation of a non-peptide, orally available FXa inhibitor, we found that a series of N-[(7-amidino-2-naphthyl)methyl]aniline derivatives possessed potent and selective inhibitory activities. Structure- activity relationship (SAR) of the substituent (R(1)) on the central aniline moiety suggested that increasing lipophilicity caused a detrimental effect on anticoagulant activity (prothrombin time assay) in plasma. Several compounds bearing a hydrophilic substituent in R(1) showed not only potent FXa inhibitory activities but also high anticoagulant activities. The best compound in this series was sulfamoylacetic acid derivative (YM-60828) which was a potent, selective and orally bioavailable FXa inhibitor and was chosen for clinical development. PMID- 11886814 TI - I(2)-imidazoline binding site affinity of a structurally different type of ligands. AB - Two families of compounds with affinity towards the I(2) imidazoline binding sites are reported. The first is a family of compounds structurally related to agmatine with two guanidine or 2-aminoimidazoline groups at each end of an aliphatic chain of six, eight, nine or 12 methylene groups. Second, and following the model of clonidine, we propose another family of compounds also with two guanidine or 2-aminoimidazoline groups at each end of a chain consisting of two phenyl rings connected by groups such as CH(2), CO, NH and SO(2). The affinity of the compounds towards the I(2) imidazoline binding sites was then evaluated in human brain tissues. In order to determine their pharmacological selectivity versus alpha(2)-adrenoceptors, the affinity for these receptors was also evaluated for the compounds with the highest affinities at I(2) imidazoline binding sites. The results obtained show that many of the compounds exhibit a considerable affinity towards the I(2) imidazoline binding sites. The aliphatic derivatives, in particular, present a very interesting selectivity for the I(2) imidazoline binding sites versus the alpha(2) adrenoceptors. To better understand these findings, mono-guanidinium analogues of the aliphatic derivatives were synthesised and tested showing poor affinity for I(2) imidazoline binding sites. The importance of these results lies in the novelty of the chemical structures studied (dicationic aliphatic compounds particularly) because they are significantly different to those of the I(2) imidazoline binding site ligands reported to date. PMID- 11886815 TI - Orally active cephalosporins. Part 4: synthesis, structure--activity relationships and oral absorption of novel 3-(4 pyrazolylmethylthio)cephalosporins with various C-7 side chains. AB - A series of 3-(4-pyrazolylmethylthio)cephalosporins with various C-7 side chains was designed, synthesized and evaluated for antibacterial activity and oral absorption in rats. Antibacterial activity against Haemophilus influenzae was markedly increased by the C-7 oxime moiety. Deamination at the 2 position of, or introduction of a substituent such as halogen or methyl to, the 5 position of the (Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino) moiety improved oral absorption. Among these compounds, FR192752 having a (Z)-2-(2-amino-5-chlorothiazol-4-yl)-2 hydroxyiminoacetamido moiety, showed potent antibacterial activity against both Gram-positive and Gram-negative bacteria including H.influenzae and penicillin G resistant Streptococcus pneumoniae (PRSP). Further, it showed higher oral absorption than CFDN and FK041. PMID- 11886816 TI - Absolute stereostructure of potent alpha-glucosidase inhibitor, Salacinol, with unique thiosugar sulfonium sulfate inner salt structure from Salacia reticulata. AB - A most potent alpha-glucosidase inhibitor named salacinol has been isolated from an antidiabetic Ayurvedic traditional medicine, Salacia reticulata WIGHT, through bioassay-guided separation. The absolute stereostructure of salacinol was determined on the basis of chemical and physicochemical evidence, which included the alkaline degradation of salacinol to 1-deoxy-4-thio-D-arabinofuranose and the X-ray crystallographic analysis, to be the unique spiro-like configuration of the inner salt comprised of 1-deoxy-4-thio-D-arabinofuranosyl sulfonium cation and 1' deoxy-D-erythrosyl-3'-sulfate anion. Salacinol showed potent inhibitory activities on several alpha-glucosidases, such as maltase, sucrase, and isomaltase, and the inhibitory effects on serum glucose levels in maltose- and sucrose-loaded rats (in vivo) were found to be more potent than that of acarbose, a commercial alpha-glucosidase inhibitor. PMID- 11886817 TI - Novel non-steroidal/non-anilide type androgen antagonists with an isoxazolone moiety. AB - 3-Substituted (Z)-4-(4-N,N-dialkylaminophenylmethylene)-5(4H)-isoxazolones and related compounds were designed and prepared as candidates for structurally novel androgen antagonists. Several compounds showed potent anti-androgenic activity as assessed by nuclear androgen receptor binding assay and growth inhibition assay using androgen-dependent Shionogi carcinoma cells SC-3. They were approximately 10--220 times more potent than flutamide in these assay systems. They also showed anti-androgenic activity toward prostate tumor cell line LNCaP, which has an aberrant nuclear androgen receptor. PMID- 11886818 TI - Synthesis of potential thrombin inhibitors. Incorporation of tartaric acid templates as P2 proline mimetics. AB - With the objective to prepare novel non-peptidic thrombin inhibitors, bioisosteres of the inhibitory tripeptide D-Phe-Pro-Arg chain have been examined. Thus, the P1 Arg was replaced with p-amidinobenzylamine, an elongated homologue of the same and with 2,5-dichloro benzylamine. The P2-P3, D-Phe-Pro, was replaced with a novel tartaric acid template coupled to a series of readily available, mainly lipophilic, amines. Some of these compounds exhibit promising thrombin inhibition activity in vitro, IC(50 ) approximately 5.9 microM. PMID- 11886819 TI - Antioxidant activity of synthetic cytokinin analogues: 6-alkynyl- and 6 alkenylpurines as novel 15-Lipoxygenase inhibitors. AB - Synthetic cytokinin analogues as well as the well known CKs 6-benzylaminopurine (BAP), kinetin and trans-zeatin were examined for antioxidant activity. The compounds were tested as potential diphenylpicrylhydrazyl (DPPH) scavengers and as inhibitors of 15-lipoxygenase (15-LO). The natural plant hormones were essentially inactive in both assays, but several synthetic analogues have a profound inhibiting effect on 15-lipoxygenase from soybeans. The same compounds were only weak DPPH scavengers and they may therefore be regarded as so-called non antioxidant inhibitors of 15-LO. PMID- 11886820 TI - Mapping of possible binding sequences of two beta-sheet breaker peptides on beta amyloid peptide of Alzheimer's disease. AB - Aggregation of amyloid peptide (Abeta) has been identified as a major feature of the pathogenesis of Alzheimer's disease. Increased risk for disease is associated with increased formation of polymerized Abeta. Inhibition of formation of toxic (aggregated) form of Abeta is one of the therapeutic possibilities. Beta sheet breaker peptides (BSBs) fulfill the requirements of an effective inhibitor. After having attached to the Abeta molecules, BSBs can prevent aggregation of Abeta to polymeric forms (aggregates). In the present study, we performed molecular modelling of complex formation between Abeta and two BSB peptides. Our aim was to find proper binding sequences for the BSB peptides on Abeta and characterize them. A dimeric model of Abeta was also used to study the interaction of BSBs with the aggregated forms of Abeta and find the sequences responsible for the polymerization process. A fast and efficient computational method: molecular docking was used for the afore-mentioned purposes. PMID- 11886821 TI - Structure--activity relationships of 1beta-methyl-2-(5-phenylpyrrolidin-3 ylthio)carbapenems. AB - Structure--activity relationship studies of 1beta-methyl-2-[(3S,5R)-5-(4 aminomethylphenyl)pyrrolidin-3-ylthio]carbapenems, especially those pertaining to the relationship between antibacterial activity and side-chain structure were conducted. These studies suggested that the trans-(3S,5R)-5-phenylpyrrolidin-3 ylthio side-chain and the aminomethyl group at the 4-position of the phenyl ring play a key role in enhancing the antibacterial activity against the MRSA and Pseudomonas aeruginosa strains. In particular, the basicity of a substituent at the 4-position of the phenyl ring were shown to greatly contribute to the antibacterial activity against MRSA and methicillin-resistant Staphyloccocus epidermidis strains. In contrast, the amidine group was shown to lead to potent antibacterial activity against P. aeruginosa strains comparable to that of imipenem, however, a good correlation between the basicity of the 4-substituent and antipseudomonal activity was not observed. In conclusion, the 4-aminomethyl or methylaminomethyl group on the phenyl ring was the best substituent for antipseudomonal activity. PMID- 11886822 TI - Benzoyl and cinnamoyl nitrogen mustard derivatives of benzoheterocyclic analogues of the tallimustine: synthesis and antitumour activity. AB - A series of benzoyl and cinnamoyl nitrogen mustards tethered to different benzoheterocycles and to oligopyrroles structurally related to netropsin consisting of two pyrrole-amide units and terminating with an amidine moiety have been synthesised and a structure--activity relationship determined. Derivatives 3 -10 have been evaluated for their sequence selective alkylating properties and cytotoxicity against human K562 leukaemia cells. They are 2- to 50-fold less cytotoxic than tallimustine, with compound 8 being the most potent member of this series. Among tallimustine isosters, the compounds with an indole 3 or benzothiophene 6 are 4-fold less cytotoxic than tallimustine, while the compounds with an N-methyl indole or benzofuran showed a 7- and 14-fold reduced cytotoxic potency, respectively. Our preliminary results indicate that these derivatives preferentially bind to AT-rich sequence with a sequence selectivity similar to tallimustine. PMID- 11886823 TI - Powerful antioxidative agents based on garcinoic acid from Garcinia kola. AB - Investigation on the structure--antioxidative activity relationships of derivatives based on garcinoic acid from Garcinia kola (Guttiferae) led to discovery of a powerful antioxidative agent. PMID- 11886824 TI - 9-Carboxymethyl-5H,10H-imidazo[1,2-a]indeno[1,2-e]pyrazin-4-one-2-carbocylic acid (RPR117824): selective anticonvulsive and neuroprotective AMPA antagonist. AB - Excessive release of glutamate, a potent excitatory neurotransmitter, is thought to play an important role in a variety of acute and chronic neurological disorders, suggesting that excitatory amino acid antagonists may have broad therapeutic potential in neurology. Here, we describe the synthesis, pharmacological properties and neuroprotective activity of 9-carboxymethyl imidazo-[1-2a]indeno[1-2e]pyrazin-4-one-2-carboxylic acid (RPR117824), an original selective AMPA antagonist. RPR117824 can be obtained through a six-step synthesis starting from (1-oxo-indan-4-yl) acetic acid, which has been validated on a gram-scale with an overall yield of 25%. Monosodium or disodium salts of the compound exhibit excellent solubility in saline (> or = 10 g/L), enabling intravenous administration. RPR117824 displays nanomolar affinity (IC(50)=18 nM) for AMPA receptors and competitive inhibition of electrophysiological responses mediated by AMPA receptors heterologously expressed in Xenopus oocytes (K(B)=5 nM) and native receptors in rat brain slices (IC(50)=0.36 microM). In in vivo testing, RPR117824 behaves as a powerful blocker of convulsions induced in mice or rats by supramaximal electroshock or chemoconvulsive agents such as pentylenetetrazole, bicuculline, isoniazide, strychnine, 4-aminopyridine and harmaline with half maximal effective doses ranging from 1.5 to 10 mg/kg following subcutaneous or intraperitoneal administration. In disease models in rats and gerbils, RPR117824 possesses significant neuroprotective activity in global and focal cerebral ischemia, and brain and spinal cord trauma. PMID- 11886827 TI - Inadequate placement of osteochondral plugs may induce abnormal stress-strain distributions in articular cartilage --finite element simulations. AB - The transplantation of osteochondral (cartilage-bone) plugs is an alternative approach to treat local, full thickness cartilage defects in young patients. It is technically difficult to control the amount of the press fit tolerance and the position of the osteochondral (OC) plug in the recipient hole. Inadequate placement of the OC plugs may produce abnormal stress and strain distributions within the cartilage, and thus influence the regeneration of the injured cartilage site and the maintenance of opposing, healthy cartilage surfaces. In the present study, the influence of press fit tolerance and the placement of the OC plug on the joint contact mechanics was simulated using finite element methods. The joint was assumed to be axi-symmetric with a spherical femur and tibia and a cylindrical OC plug. Our simulations showed that small misplacements of the OC plug induced abnormal tension in the articular cartilage of the opposing, healthy cartilage surface. Such tension might induce unpredictable adaptations, or possibly degenerations, in the opposing cartilage layer. The contact stress profiles in the joint were predicted to change discontinuously across the plug/recipient interface, even when the plug was perfectly placed in the recipient hole, i.e., the plug's surface was aligned with the recipient surface. For a fixed coefficient of friction and a fixed fit tolerance, the maximal sliding force was predicted to vary with the size of the plug and reached a maximum at a specific plug diameter. The present simulations should be helpful for the design of instruments for osteochondral transplantation and placement of OC plugs, for understanding articular cartilage adaptation following osteochondral repair, and for providing insight into the mechanics at the transplant/recipient interface where proper integration of the plug into the joint is most problematic. PMID- 11886825 TI - Novel Tn antigen-containing neoglycopeptides: synthesis and evaluation as anti tumor vaccines. AB - The fully unprotected alpha-C-glycosyl analogue of N-acetylgalactosamine 9 was conjugated by a non-natural oxime bond to the segment peptides (328--340)OVA and (327--339)OVA, affording neoglycopeptides 1--2 and 3, having one or two sugar units, respectively. The three neoglycopeptides were tested in vitro in an antigen presentation assay as antitumor vaccines. Neoglycopeptides 1--3 could be presented to and recognized by the T cell receptor; neoglycopeptide 3, bearing two B-epitopes, was presented to the TCR with higher efficiency, compared to neoglycopeptide 2, having only one B-epitope. PMID- 11886828 TI - Importance of the superficial tissue layer for the indentation stiffness of articular cartilage. AB - Indentation testing is a widely used technique for nondestructive mechanical analysis of articular cartilage. Although cartilage shows an inhomogeneous, layered structure with anisotropic mechanical properties, most theoretical indentation models assume material homogeneity and isotropy. In the present study, quantitative polarized light microscopy (PLM) measurements from canine cartilage were utilized to characterize thickness and structure of the superficial, collageneous tissue layer as well as to reveal its relation to experimental indentation measurements. In addition to experimental analyses, a layered, transversely isotropic finite element (FE) model was developed and the effect of superficial (tangential) tissue layer with high elastic modulus in the direction parallel to articular surface on the indentation response was studied. The experimental indentation stiffness was positively correlated with the relative thickness of the superficial cartilage layer. Also the optical retardation, which reflects the degree of parallel organization of collagen fibrils as well as collagen content, was related to indentation stiffness. FE results indicated effective stiffening of articular cartilage under indentation due to high transverse modulus of the superficial layer. The present results suggest that indentation testing is an efficient technique for the characterization of the superficial degeneration of articular cartilage. PMID- 11886829 TI - The influence of design parameters on cortical strain distribution of a cementless titanium femoral stem. AB - The strain distribution imposed on a femur following a total joint replacement is an important factor, in proximal bone loss due to stress shielding, and long term clinical success. This study investigated how five different design parameters of a cementless titanium femoral prosthesis influenced cortical strains. Test loads were applied and strains were measured with and without an abductor force simulation, using six human cadaveric femora. The cementless design used demonstrated significant calcar loading proximally and a similar strain distribution to the intact femur distally. Implant gross geometry was the major factor in determining the cortical strain distributions under abductor simulation in both axial and torsional loading. PMID- 11886830 TI - Motion quality evaluation of upper limb target-reaching movements. AB - Fitts' Law was extended in the polar coordinate system, and a set of indices for human motion evaluation is proposed. In this paper, the index of difficulty and the index of performance are introduced as the general indices for the quality measure of plane target-to-target movement. As an example, the target-reaching movement of the upper limb, which is a basic functional action of upper limbs in the activities of daily living, was experimentally investigated. Five healthy subjects were asked to perform six target-reaching tasks with different indices of difficulty. All movements were recorded using a Vicon motion analysis system. The movement quality was measured using these evaluation indices. PMID- 11886831 TI - A dynamic model for simulating a trip and fall during gait. AB - The purpose of this study was to develop an analytical model to simulate a trip and fall during gait. The human body was modeled as a 12 degree-of-freedom linkage system. The kinematics of the lower extremity for one cycle of gait were obtained for a healthy subject using an optoelectronic three-dimensional data acquisition system. Inverse dynamics was used to compute the moments about the hip, knee and ankle joints of the lower extremity. These moments were then used as input actuators to the joints in to a forward dynamics model to simulate the swing phase of gait from toe-off to heel-strike. An optimization procedure to minimize errors associated with the computed experimental torque was applied to correct for mathematical instability. An experiment was performed to measure the three-dimensional foot--obstacle contact force for a healthy subject tripping on an obstacle during gait. The contact force was applied to the swing limb of the forward dynamics model for 0.09 s beginning at 0.04 s after toe-off. Tripping on an obstacle followed by a muscle-relaxed fall was simulated. The simulation results were visualized with animation software. PMID- 11886832 TI - Lacunarity analysis of spatial pattern in CT images of vertebral trabecular bone for assessing osteoporosis. AB - The structural integrity of vertebral trabecular bone is determined by the continuity of its trabecular network and the size of the holes comprising its marrow space, both of which determine the apparent size of the marrow spaces in a transaxial CT image. A model-independent assessment of the trabeculation pattern was determined from the lacunarity of thresholded CT images. Using test images of lumbar vertebrae from human cadavers, acquired at different slice thicknesses, we determined that both median thresholding and local adaptive thresholding (using a 7 x 7 window) successfully segmented the grey-scale images. Lacunarity analysis indicated a multifractal nature to the images, and a range of marrow space sizes with significant structure around 14-18 mm(2). Preliminary studies of in vivo images from a clinical CT scanner indicate that lacunarity analysis can follow the pattern of bone loss in osteoporosis by monitoring the homogeneity of the marrow spaces, which is related to the connectivity of the trabecular bone network and the marrow space sizes. Although the patient sample was small, derived parameters such as the maximum deviation of the lacunarity from a neutral (fractal) model, and the maximum derivative of this deviation, seem to be sufficiently sensitive to distinguish a range of bone conditions. Our results suggest that these parameters, used with bone mineral density values, may have diagnostic value in characterizing osteoporosis and predicting fracture risk. PMID- 11886833 TI - Assessment of the ultrasonic dental scaler insert. AB - The ultrasonic dental scaler is an invaluable tool in the fight against dental calculus and periodontal disease. The purpose of this study was to assess the natural frequency of the straight internal ultrasonic scaler insert 30K FSI-SLI 10S (Dentsply International, York, USA) and to measure the displacement observed at the tip when operated under different generator settings. Two approaches were used: finite element analysis and scanning laser vibrometry. The Finite Element Analysis method was employed to calculate the expected modes of vibration of the insert. Scanning laser vibrometry was used to measure displacements of the tip during operation. It was found the insert had two modes of vibration around the operating frequency of 30 kHz; these modes were first order bending in two orthogonal planes. PMID- 11886834 TI - Assessment of a three-dimensional robotic model for biomechanical-data acquisition of human movement. AB - The use of a kinematic robotic model has not been implemented in the biomechanical-data acquisition protocol, as it has in workplace analysis, ergonomics and design. The purpose of this paper was to assess the use of a kinematic model to retrieve frames of human movements from data obtained at a low sampling frequency. From experimental trials with an original sampling frequency of 60 Hz, the data were sampled again at two lower frequencies, 5 Hz and 10 Hz. The model was then used to reconstitute the data to its original frequency (60 Hz). The results demonstrated that it was possible to retrieve a full 3-D human movement from a sampling rate lower than normal without sacrificing accuracy. It was observed from both reduced sampling frequencies that the error level was comparable to the usual accuracy of a DLT 3-D reconstruction technique. It was therefore concluded that the data retrieved from these two frequencies were very similar to the original data sampled at 60 Hz. PMID- 11886835 TI - A numerical method for estimating blood flow by dynamic functional imaging. AB - We present a numerical deconvolution scheme for estimating regional blood flow and tissue retention functions by dynamic functional imaging. The present approach implements the Tikhonov-Miller regularization in general form, which allows for prior knowledge or assumptions to be incorporated during the deconvolution process, so as to stabilize the solution against variations due to noise. Appropriate approximations and simplifications in the context of functional imaging, were also introduced to ease numerical computations. Monte Carlo simulation experiments were carried out to study the applicability of the present approach and to compare with other deconvolution techniques previously studied. PMID- 11886836 TI - An investigation of the repeatability and reproducibility of ISO 11948-1 (the Rothwell method) for measuring the absorption capacity of incontinence pads. AB - The repeatability and reproducibility (precision within and between laboratories, respectively) of an international standard method (ISO 11948-1, the Rothwell method) for measuring the absorption capacity of incontinence pads was investigated. The 74 shaped disposable bodyworn insert pads for heavy incontinence on the UK market in spring 1997 were tested in three laboratories experienced in using the method, one in each of England, Spain and Sweden. Coefficients of variation (standard deviation as a proportion of the mean) for five repeats rarely exceeded 5% within any laboratory. However, there were systematic differences between laboratories: results from the Swedish and Spanish laboratories typically exceeded those from the English laboratory by 13% and 8%, respectively. The good repeatability suggests that the method is capable of adequate precision but the poor reproducibility implies that the instructions in the standard for building and/or using the test apparatus are inadequate, leaving too much room for interpretation. Having studied the data presented here and viewed videos of the apparatus in use in five laboratories (including the three contributing to this note) the ISO working group which wrote the original standard has identified several likely sources of imprecision and is now working to revise the standard to improve its reproducibility. PMID- 11886837 TI - Complex glycosylation of Skp1 in Dictyostelium: implications for the modification of other eukaryotic cytoplasmic and nuclear proteins. AB - Recently, complex O-glycosylation of the cytoplasmic/nuclear protein Skp1 has been characterized in the eukaryotic microorganism Dictyostelium. Skp1's glycosylation is mediated by the sequential action of a prolyl hydroxylase and five conventional sugar nucleotide-dependent glycosyltransferase activities that reside in the cytoplasm rather than the secretory compartment. The Skp1-HyPro GlcNAcTransferase, which adds the first sugar, appears to be related to a lineage of enzymes that originated in the prokaryotic cytoplasm and initiates mucin-type O-linked glycosylation in the lumen of the eukaryotic Golgi apparatus. GlcNAc is extended by a bifunctional glycosyltransferase that mediates the ordered addition of beta1,3-linked Gal and alpha1,2-linked Fuc. The architecture of this enzyme resembles that of certain two-domain prokaryotic glycosyltransferases. The catalytic domains are related to those of a large family of prokaryotic and eukaryotic, cytoplasmic, membrane-bound, inverting glycosyltransferases that modify glycolipids and polysaccharides prior to their translocation across membranes toward the secretory pathway or the cell exterior. The existence of these enzymes in the eukaryotic cytoplasm away from membranes and their ability to modify protein acceptors expose a new set of cytoplasmic and nuclear proteins to potential prolyl hydroxylation and complex O-linked glycosylation. PMID- 11886838 TI - Ectopic localizations of Golgi glycosyltransferases. AB - Glycosyltransferases involved in N- and O-glycan chain elongation and termination are localized in the Golgi apparatus. Early evidence in support of this rule was based on fractionation techniques and was corroborated by numerous immunocytochemical studies. Usually these studies were confined to cultured cell lines exhibiting little differentiation features, such as HeLa cells. However, localization studies conducted in primary cell cultures (e.g., human umbilical vein endothelial cells), cells obtained ex vivo (e.g., sperm cells), and tissue sections (e.g., intestinal, renal, or hepatic tissue) often reveal ectopic localizations of glycosyltransferases usually at post-Golgi sites, including the plasma membrane. Hence, extracellular cues resulting from specific adhesion sites may influence post-Golgi trafficking routes, which may be reflected by ectopic localization of Golgi enzymes. PMID- 11886839 TI - Purification and characterization of N-acetylneuraminic acid-9-phosphate synthase from rat liver. AB - Sialic acids are a group of carboxylated amino sugars important for a variety of cellular functions. N-Acetylneuraminic acid (Neu5Ac) is the predominant sialic acid in nature. Neu5Ac-9-phosphate synthase catalyzes the formation of Neu5Ac-9 phosphate from N-acetylmannosamine-6-phosphate and phosphoenolpyruvate. Neu5Ac-9 phosphate synthase was purified 11,700-fold from rat liver cytosol to apparent homogeneity by ammonium sulfate precipitation, chromatography on hydroxylapatite, phenyl-Sepharose, MonoQ, and finally gel filtration. SDS-PAGE and gel filtration chromatography indicated that the enzyme is a dimer composed of 37-kDa subunits. Analysis of trypic peptides by MALDI-TOF MS verified a high sequence similarity to the corresponding murine enzyme. The K(m) values of Neu5Ac-9-phosphate synthase were 35 microM for N-acetylmannosamine-6-phosphate and 100 microM for phosphoenolpyruvate. The enzyme displayed an absolute requirement for divalent cations, Mn(2+), Fe(2+), and Mg(2+) being the most effective. In contrast to human Neu5Ac-9-phosphate synthase, the rat enzyme did not utilize mannose-6 phosphate in the synthesis of 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid 9 phosphate. Neu5Ac-9-phosphate synthase was inactivated by the sulfhydryl modifying reagents, 5,5'-dithio-bis (2-nitrobenzoic acid) and N-ethylmaleimide, and protected from inactivation by the presence of the substrate phosphoenolpyruvate, but not by the presence of N-acetylmannosamine-6-phosphate, showing that at least one cysteine residue is located in the active site of the enzyme. PMID- 11886840 TI - Expression of a functional Drosophila melanogaster N-acetylneuraminic acid (Neu5Ac) phosphate synthase gene: evidence for endogenous sialic acid biosynthetic ability in insects. AB - In this study, we report the first cloning and characterization of a N acetylneuraminic acid phosphate synthase gene from Drosophila melanogaster, an insect in the protostome lineage. The gene is ubiquitously expressed at all stages of Drosophila development and in Schneider cells. Similar to the human homologue, the gene encodes an enzyme with dual substrate specificity that can use either N-acetylmannosamine 6-phosphate or mannose 6-phosphate to generate phosphorylated forms of both the sialic acids, N-acetylneuraminic acid and 2-keto 3-deoxy-D-glycero-D-galacto-nononic acid, respectively, when expressed in either bacterial or baculoviral expression systems. The identification of a functional sialic acid synthase in Drosophila indicates that insects have the biosynthetic capability to produce sialic acids endogenously. Although sialylation is widely distributed in organisms of the deuterstome lineage, genetic evidence concerning the presence or absence of sialic acid metabolism in organisms of the protostome lineage has been lacking. Homology searches of the Drosophila genome identified putative orthologues of other genes required for sialylation of glycoconjugates. PMID- 11886841 TI - Purification, characterization, and cDNA cloning of alpha-N-acetylgalactosamine specific lectin from starfish, Asterina pectinifera. AB - We report here the purification, characterization, and cDNA cloning of a novel N acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. The purified lectin showed 19-kDa, 41-kDa, and 60-kDa protein bands on SDS-PAGE, possibly corresponding to a monomer, homodimer, and homotrimer. Interestingly, on 4-20% native PAGE the lectin showed at least nine protein bands, among which oligomers containing six to nine subunits had potent hemagglutination activity for sheep erythrocytes. The hemagglutination activity of the lectin was specifically inhibited by N-acetylgalactosamine, Tn antigen, and blood group A trisaccharide, but not by N-acetylglucosamine, galactose, galactosamine, or blood group B trisaccharide. The specificity of the lectin was further examined using various glycosphingolipids and biotin-labeled lectin. The lectin was found to bind to Gb5Cer, but not Gb4Cer, Gb3Cer, GM1a, GM2, or asialo-GM2, indicating that the lectin specifically binds to the terminal alpha-GalNAc at the nonreducing end. The hemagglutination activity of the lectin was completely abolished by chelation with EDTA or EGTA and completely restored by the addition of CaCl(2). cDNA cloning of the lectin showed that the protein is composed of 168 amino acids, including a signal sequence of 18 residues, and possesses the typical C type lectin motif. These findings indicate that the protein is a C-type lectin. The recombinant lectin, produced in a soluble form by Escherichia coli, showed binding activity for asialomucin in the presence of Ca(2+) but no hemagglutination. PMID- 11886842 TI - Glycosylation of the hepatitis C virus envelope protein E1 occurs posttranslationally in a mannosylphosphoryldolichol-deficient CHO mutant cell line. AB - The addition of N-linked glycans to a protein is catalyzed by oligosaccharyltransferase, an enzyme closely associated with the translocon. N glycans are believed to be transferred as the protein is being synthesized and cotranslationally translocated in the lumen of the endoplasmic reticulum. We used a mannosylphosphoryldolichol-deficient Chinese hamster ovary mutant cell line (B3F7 cells) to study the temporal regulation of N-linked core glycosylation of hepatitis C virus envelope protein E1. In this cell line, truncated Glc(3)Man(5)GlcNAc(2) oligosaccharides are transferred onto nascent proteins. Pulse-chase analyses of E1 expressed in B3F7 cells show that the N-glycosylation sites of E1 are slowly occupied until up to 1 h after protein translation is completed. This posttranslational glycosylation of E1 indicates that the oligosaccharyltransferase has access to this protein in the lumen of the endoplasmic reticulum for at least 1 h after translation is completed. Comparisons with the N-glycosylation of other proteins expressed in B3F7 cells indicate that the posttranslational glycosylation of E1 is likely due to specific folding features of this acceptor protein. PMID- 11886843 TI - Specific effects of fructo- and gluco-oligosaccharides in the preservation of liposomes during drying. AB - The fructan family of oligo- and polysaccharides is a group of molecules that have long been implicated as protective agents in the drought and freezing tolerance of many plant species. However, it has been unclear whether fructans have properties that make them better protectants for cellular structures than other sugars. We compared the effects of fructans and glucans on membrane stability during air-drying. Although glucans of increasing chain length were progressively less able to stabilize liposomes against leakage of aqueous content after rehydration, fructans showed increased protection. On the other hand, glucans became more effective in protecting liposomes against membrane fusion with increasing chain length, whereas fructans became less effective. Fourier transform infrared spectroscopy showed a reduction of the gel to liquid crystalline phase transition temperature (T(m)) of air-dried liposomes by approximately 25 degrees C in the presence of sucrose and maltose. For the respective pentasaccharides, the reduction of T(m) of the lipids was 9 degrees C lower for samples containing fructan than for those containing glucan, indicating increased sugar--membrane interactions for the fructan compared to the glucan. A reduced interaction of the longer-chain glucans and an increased interaction of the respective fructans with the phospholipid head groups in the dry state was also indicated by dramatic differences in the phosphate asymmetric stretch region of the infrared spectrum. Collectively, our data indicate that the fructo oligosaccharides accumulated in many plant species under stress conditions could indeed play an important role in cellular dehydration tolerance. PMID- 11886844 TI - Regulation of galectin-9 expression and release in Jurkat T cell line cells. AB - Ecalectin/galectin-9 was recently described as a novel eosinophil chemoattractant highly expressed in immune tissues. We investigated the regulation of galectin-9 expression and release in Jurkat (a T cell line) cells. We demonstrated that medium and long-sized galectin-9 isoforms were constitutively expressed, and phorbol 12-myriastate 13-acetate (PMA) upregulated the level of galectin-9 mRNA in Jurkat cells. Western blotting and flow cytometry analyses revealed that PMA stimulation resulted in the upregulation of both intracellular and surface galectin-9 protein. The stimulated Jurkat cells simultaneously released evident eosinophil chemoattractant activity (ECA). Main ECA was adsorbed by both lactose and anti-galectin-9 antibody affinity column, suggesting that the ECA was ascribed to galectin-9. When Jurkat cells were stimulated with PMA in the presence of a BB94, a matrix metalloproteinase (MMP) inhibitor, but not tissue inhibitor of metalloproteinase-1 (TIMP-1), the release of galectin-9 was suppressed in a dose-dependent manner. We further found that calphostin c, a protein kinase c (PKC) inhibitor, weakly but significantly suppressed the release of galectin-9. The present data suggested that galectin-9 production in Jurkat cells is provoked by the stimulation with PMA and that some MMP and PKC is, at least, partly involved in the release of galectin-9 from Jurkat cells. PMID- 11886845 TI - UDP-GlcNAc concentration is an important factor in the biosynthesis of beta1,6 branched oligosaccharides: regulation based on the kinetic properties of N acetylglucosaminyltransferase V. AB - Human beta1,6-N-acetylglucosaminyltransferase V (GnT-V) was expressed by baculovirus-insect cell system, and the purified recombinant enzyme was kinetically characterized. The data obtained were used to establish the kinetic basis of the substrate specificity toward donor nucleotide sugars, and also revealed that K(m) values for the donors are much higher compared to those of other GlcNAc transferases, the kinetic properties of which have been reported. Because this exceptionally higher K(m) suggests that GnT-V is physiologically present at far from saturated conditions, it would appear that the production of beta1,6-branched oligosaccharide, which is formed by GnT-V, could be regulated in vivo by the concentration of the donor, UDP-GlcNAc, as well as the expression levels of the enzyme. When B16 melanoma cells, which express high levels of GnT V, were incubated with GlcNAc, the beta1,6-branched oligosaccharide levels were increased, as judged by a lectin blot analysis, in conjunction with an increase in intracellular UDP-GlcNAc. These findings suggest that the level of UDP-GlcNAc can be a critical factor in the production of beta1,6-branched oligosaccharides, for example, by tumor cells, which have been thought to be closely associated with tumor progression and metastasis. PMID- 11886846 TI - Galectin-3 is strongly up-regulated in nonapoptosing mammary epithelial cells during rat mammary gland involution. AB - Galectin-3 is an endogenous mammalian lectin that binds to ABH carbohydrate antigens. Here we show that galectin-3 is strongly up-regulated during mammary gland involution and that it is expressed virtually exclusively on nonapoptotic cells. We demonstrate that dexamethasone, an inhibitor of the second phase of mammary gland involution, potently suppresses up-regulation of galectin-3 as judged immunohistochemically and on western blots, suggesting that systemic hormone levels regulate galectin-3 expression during involution. However, at the RNA level galectin-3 expression is rapidly up-regulated on the onset of involution but remains consistantly high during the first and second phase of involution regardless of dexamethasone treatment. These data suggest that the up regulation of galectin-3 in the involuting mammary gland is not only controlled transcriptionally but also regulated posttranscriptionally under the control of systemic glucocorticoid hormones involved in coordinating the involution process. PMID- 11886847 TI - Expression of alpha-gal epitopes on HeLa cells transduced with adenovirus containing alpha1,3galactosyltransferase cDNA. AB - Alpha1,3galactosyltransferase (alpha1,3GT) synthesizes alpha-gal epitopes (Gal(alpha)1-3Galbeta1-4GlcNAc-R) on glycoconjugates in nonprimate mammals but not in humans. Transduction of alpha1,3GT gene into human HeLa cells by an adenovirus vector allowed for accurate kinetics studies on the appearance of alpha1,3GT and of its product, the alpha-gal epitope, in the transduced cells. Mouse alpha1,3GT cDNA was inserted into a replication-defective adenovirus vector. This viral vector, designated Ad(alpha)GT, could be propagated in human 293 cells that have the viral E1 complementing gene. Transduction of HeLa cells resulted in immediate penetration of approximately 20 Ad(alpha)GT copies into each cell and the appearance of alpha1,3GT mRNA after 4h. Catalytic activity of alpha1,3GT was first detected in the cells after 6 h. The initial appearance of alpha-gal epitopes (approximately 6 x 10(4)/cell) on cell surface glycoconjugates was detected 10 h posttransduction, whereas 24 h posttransduction each cell expressed 2 x 10(6) epitopes. The activity of alpha1,3GT in cells transduced with approximately two copies of Ad(alpha)GT was eightfold lower than that in cells transduced with approximately 20 Ad(alpha)GT copies; however, the number of alpha gal epitopes/cell remained closely similar. This implies that increased alpha1,3GT activity above a certain saturation level does not result in a corresponding increase in the carbohydrate product, possibly because of competing glycosyltransferases. PMID- 11886848 TI - CD40 induces interleukin-6 gene transcription in dendritic cells: regulation by TRAF2, AP-1, NF-kappa B, AND CBF1. AB - CD40-induced activation of cytokine gene expression in dendritic cells (DC) is an important process in the initiation of primary immune responses. We have determined the intracellular signaling events that lead to CD40 ligation-induced activation of interleukin-6 (IL-6) gene transcription in a murine DC line, FSDC, that is phenotypically representative of bone marrow-derived DC. IL-6 reverse transcriptase-PCR and promoter assays established the responsiveness of FSDC to anti-CD40 ligation. Further promoter assays showed that the transcription factors NF-kappaB and AP-1 are downstream transcriptional mediators of CD40-induced IL-6 gene expression. Anti-CD40 treatment of FSDC stimulated increased expression of specific NF-kappaB (p50:p65) and AP-1 (c-Jun, JunB, JunD, and c-Fos) DNA-protein complexes. Overexpression of an IkappaB-alpha super-repressor or a dominant negative JunD resulted in a strong inhibition of CD40-inducible IL-6 promoter activity supporting a role for both transcription factors. Upstream signal transduction events were studied by transfection of wild type and mutant human CD40 expression constructs into FSDC followed by stimulation with an anti-human CD40 antibody. These experiments revealed that anti-CD40 stimulation of NF-kappaB and IL-6 gene transcription requires specific amino acid residues in the cytoplasmic region of CD40 involved in the recruitment of TRAF2. Induction of IL 6 mRNA by anti-CD40 treatment was found to be a transient event (24 h) and was followed by a diminution of IL-6 transcript to levels below those found in unstimulated cells. This loss of IL-6 expression was associated with reduced p50:p65 NF-kappaB DNA binding and elevated binding of CBF1 to a site overlapping the NF-kappaB site. Overexpression of CBF1 resulted in a profound inhibition of basal and anti-CD40-induced IL-6 promoter activities indicating that prolonged induction of CBF1 may contribute to the transient nature of the IL-6 response. The physiological relevance of these molecular events to DC function is discussed. PMID- 11886849 TI - New alternatively spliced form of galectin-3, a member of the beta-galactoside binding animal lectin family, contains a predicted transmembrane-spanning domain and a leucine zipper motif. AB - Osteoclasts or their precursors interact with the glycoprotein-enriched matrix of bone during extravasation from the vasculature, and upon attachment prior to resorption. Reverse transcriptase-PCR studies showed that two new alternatively spliced forms of chicken galectin-3, termed Gal-3TM1 and Gal-3TR1, were enriched and preferentially expressed in highly purified chicken osteoclast-like cells. Gal-3TM1 and Gal-3TR1 mRNA were also detected in chicken intestinal tissue, but not in kidney, liver, or lung. Gal-3TM1 and Gal-3TR1 messages both contain an open reading frame encoding a predicted 70-amino acid TM1 sequence inserted between the N-terminal Gly/Pro repeat domain and the carbohydrate recognition domain (exons 3 and 4). Gal-3TR1 mRNA contains an additional 241-bp sequence, which encodes a truncated open reading frame between the 4th and 5th exons, and, whose translation is expected to terminate within the carbohydrate recognition domain encompassing exons 4, 5, and 6. Immunoblotting and affinity chromatography showed that purified osteoclast preparations and intestinal homogenates contained a 36-kDa lactose-binding galectin. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analyses on chymotryptic peptides from the 36 kDa lectin confirmed its identity as Gal-3TM1. The TM1 insert contains a single transmembrane-spanning region and a leucine zipper-like stalk domain that is predicted to position the intact carbohydrate recognition domain of Gal-3TM1 on the exterior surface of the plasma membrane. Immunofluorescent staining of chicken osteoclasts confirmed the expression of Gal-3TM1 at the plasma membrane. Gal-3TM1 is the first example of a galectin superfamily member capable of being expressed as a soluble protein and as a transmembrane protein. PMID- 11886850 TI - Cytoplasmic localization of tristetraprolin involves 14-3-3-dependent and independent mechanisms. AB - The immediate early gene tristetraprolin (TTP) is induced transiently in many cell types by numerous extracellular stimuli. TTP encodes a zinc finger protein that can bind and destabilize mRNAs that encode tumor necrosis factor-alpha (TNFalpha) and other cytokines. We hypothesize that TTP also has a broader role in growth factor-responsive pathways. In support of this model, we have previously determined that TTP induces apoptosis through the mitochondrial pathway, analogously to certain oncogenes and other immediate-early genes, and that TTP sensitizes cells to the pro-apoptotic signals of TNFalpha. In this study, we show that TTP and the related proteins TIS11b and TIS11d bind specifically to 14-3-3 proteins and that individual 14-3-3 isoforms preferentially bind to different phosphorylated TTP species. 14-3-3 binding does not appear to inhibit or promote induction of apoptosis by TTP but is one of multiple mechanisms that localize TTP to the cytoplasm. Our results provide the first example of 14-3-3 interacting functionally with an RNA binding protein and binding in vivo to a Type II 14-3-3 binding site. They also suggest that 14-3-3 binding is part of a complex network of stimuli and interactions that regulate TTP function. PMID- 11886852 TI - Regulation of phospholipase C-gamma activity by glycosphingolipids. AB - Glycosphingolipid-enriched domains are hot spots for cell signaling within plasma membranes and are characterized by the enrichment of glycosphingolipids. A role for glucosylceramide-based glycosphingolipids in phospholipase C-mediated inositol 1,4,5-trisphosphate formation has been previously documented. These earlier studies utilized a first generation glucosylceramide synthase inhibitor to deplete cells of their glycosphingolipids. Recently, more active and specific glucosylceramide synthase inhibitors, including d-threo-ethylendioxyphenyl-2 palmitoylamino-3-pyrrolidinopropanol (d-t-EtDO-P4), have been designed. d-t-EtDO P4 has the advantage of blocking glucosylceramide synthase at low nanomolar concentrations but does not cause secondary elevations in cell ceramide levels. In the present study, d-t-EtDO-P4 depleted cellular glucosylceramide and lactosylceramide in cultured ECV304 cells at nanomolar concentrations without obvious cellular toxicity. The expression of several signaling proteins was evaluated in glycosphingolipid-depleted ECV304 cells to study the role of glycosphingolipids in phospholipase C-mediated signaling. No difference was observed in the cellular expression of phospholipase C-gamma between controls and glycolipid-depleted cells. Western blot analysis, however, revealed that depletion of endogenous glycosphingolipids in cultured ECV304 cells with d-t-EtDO P4 induced tyrosine phosphorylation of phospholipase C-gamma in a concentration dependent manner with maximum induction at 100 nm. The phosphorylation of phospholipase C-gamma induced by d-t-EtDO-P4 was abolished by exogenously added glucosylceramide, consistent with a specific glycosphingolipid-phospholipase C gamma interaction. The phospholipase C-gamma phosphorylation was maximally enhanced by bradykinin when cells were exposed to 100 nm d-t-EtDO-P4. The measurement of cellular activity of phospholipase C-gamma, by myo-inositol 1,4,5 trisphosphate radioreceptor assay, demonstrated that depletion of glucosylceramide-based glycosphingolipids in cultured ECV304 cells with d-t-EtDO P4 resulted in significantly increased formation of inositol 1,4,5-trisphosphate above base line, and an increased sensitivity of phospholipase C-gamma to bradykinin stimulation. Thus, the activation of phospholipase C-gamma is negatively regulated by membrane glycosphingolipids in ECV304 cells. PMID- 11886851 TI - Interaction of elongation factor-1alpha and pleckstrin homology domain of phospholipase C-gamma 1 with activating its activity. AB - The pleckstrin homology (PH) domain is a small motif for membrane targeting in the signaling molecules. Phospholipase C (PLC)-gamma1 has two putative PH domains, an NH(2)-terminal and a split PH domain. Here we report studies on the interaction of the PH domain of PLC-gamma1 with translational elongation factor (EF)-1alpha, which has been shown to be a phosphatidylinositol 4-kinase activator. By pull-down of cell extract with the glutathione S-transferase (GST) fusion proteins with various domains of PLC-gamma1 followed by peptide sequence analysis, we identified EF-1alpha as a binding partner of a split PH domain of PLC-gamma1. Analysis by site-directed mutagenesis of the PH domain revealed that the beta2-sheet of a split PH domain is critical for the interaction with EF 1alpha. Moreover, Dot-blot assay shows that a split PH domain specifically binds to phosphoinositides including phosphatidylinositol 4-phosphate and phosphatidylinositol 4, 5-bisphosphate (PIP(2)). So the PH domain of PLC-gamma1 binds to both EF-1alpha and PIP(2). The binding affinity of EF-1alpha to the GST.PH domain fusion protein increased in the presence of PIP(2), although PIP(2) does not bind to EF-1alpha directly. This suggests that EF-1alpha may control the binding affinity between the PH domain and PIP(2). PLC-gamma1 is substantially activated in the presence of EF-1alpha with a bell-shaped curve in relation to the molar ratio between them, whereas a double point mutant PLC-gamma1 (Y509A/F510A) that lost its binding affinity to EF-1alpha shows basal level activity. Taken together, our data show that EF-1alpha plays a direct role in phosphoinositide metabolism of cellular signaling by regulating PLC-gamma1 activity via a split PH domain. PMID- 11886853 TI - Protein kinase A deficiency causes axially localized neural tube defects in mice. AB - We have studied the function of protein kinase A (PKA) during embryonic development using a PKA-deficient mouse that retains only one functional catalytic subunit allele, either Calpha or Cbeta, of PKA. The reduced PKA activity results in neural tube defects that are specifically localized posterior to the forelimb buds and lead to spina bifida. The affected neural tube has closed appropriately but exhibits an enlarged lumen and abnormal neuroepithelium. Decreased PKA activity causes dorsal expansion of Sonic hedgehog signal response in the thoracic to sacral regions correlating with the regions of morphological abnormalities. Other regions of the neural tube appear normal. The regional sensitivity to changes in PKA activity indicates that downstream signaling pathways differ along the anterior-posterior axis and suggests a functional role for PKA activation in neural tube development. PMID- 11886854 TI - Tryptophan 1093 is largely responsible for the slow off rate of calmodulin from plasma membrane Ca2+ pump 4b. AB - Tryptophan 1093 resides in the 28-residue calmodulin-binding/autoinhibitory domain of the plasma membrane Ca(2+) pump (PMCA). Previous studies with the isolated calmodulin-binding/autoinhibitory peptide from PMCA have shown that mutations of the tryptophan residue decrease the affinity of the peptide for calmodulin and its affinity as an inhibitor of proteolytically activated pump. In this study, the PMCA mutation in which tryptophan 1093 is converted to alanine (W1093A) was constructed in the full-length PMCA isoform 4b. The mutant pump was expressed in COS cells, and its steady state and pre-steady state kinetic properties were examined. The W1093A pump exhibited an increased basal activity in the absence of calmodulin, so the activation was approximately 2-fold (it is 10-fold in the wild type). The W1093A mutation also lowered the steady state affinity for calmodulin from K(0.5) of 9 nm for wild type to 144 nm (assayed at 700 nm free Ca(2+)). Pre-steady state measurements of the rate of activation by Ca(2+)-calmodulin revealed that the W1093A mutant responded 2.5-fold faster to calmodulin. In contrast to these relatively modest effects, the half-time of inactivation of the mutant was reduced by more than 2 orders of magnitude from 41 min to 7 s. We conclude that tryptophan 1093 does not play a substantial role in Ca(2+)-calmodulin recognition; rather it functions primarily to slow the inactivation of the calmodulin-activated pump. PMID- 11886855 TI - Computer modeling of three-dimensional structure of DNA-packaging RNA (pRNA) monomer, dimer, and hexamer of Phi29 DNA packaging motor. AB - A striking common feature in the maturation of all linear double-stranded DNA viruses is that their lengthy genome is translocated with remarkable velocity into the limited space within a preformed protein shell and packaged into near crystalline density. A DNA-translocating motor, powered by ATP hydrolysis, accomplishes this task, which would otherwise be energetically unfavorable. DNA packaging RNA, pRNA, forms a hexameric complex to serve as a vital component of the DNA translocating motor of bacterial virus Phi29. The sequential action of six pRNA ensures continual function in the DNA translocation process. The Phi29 motor has been assembled with purified components synthesized by chemical or biotechnological approaches and is able to pump the viral DNA into the protein shell in vitro. pRNA dimers are the building blocks of the hexamer. The computer models of the three-dimensional structure of the motor was constructed based on experimental data derived from photoaffinity cross-linking by psoralen, phenphi (cis-Rh(1,10-phenanthroline)(9,10-phenan-threnequinone diimine)Cl(2)(+)), and azidophenacyl; chemical modification and chemical modification interference with dimethyl sulfate, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p-toluene sulfonate, and kethoxal; complementary modification; and nuclease probing by single- and double-stranded specific RNases. The shapes of these computer models are very similar to the published pRNA images of cryo-atomic force microscopy. pRNA hexamer docking with the connector crystal structure reveals a very impressive match with the available biochemical, genetic, and physical data. PMID- 11886856 TI - cAMP-dependent protein kinase types I and II differentially regulate cAMP response element-mediated gene expression: implications for neuronal responses to ethanol. AB - We have shown that ethanol induces translocation of cAMP-dependent protein kinase (PKA) to the nucleus, cAMP response element-binding protein (CREB) phosphorylation, and cAMP response element-mediated gene transcription in NG108 15 cells. However, little is known about which PKA types regulate this process. We show here that under basal conditions NG108-15 cells contain type I PKA (CbetaRIbeta) primarily in cytosol and type II PKA (CalphaRIIbeta) in the particulate and nuclear fractions. Antagonists of both type I and type II PKA inhibit forskolin- and ethanol-induced cAMP response element-mediated gene transcription. However, only the type II PKA antagonist inhibits forskolin induced Calpha and ethanol-induced Calpha and RIIbeta translocation to the nucleus and CREB phosphorylation; the type I antagonist is without effect. Our data suggest that forskolin- and ethanol-induced CREB phosphorylation and gene activation are differentially mediated by the two types of PKA. We propose that type II PKA is translocated and activated in the nucleus and induces CREB phosphorylation that is necessary but not sufficient for gene transcription. By contrast, type I PKA is activated in the cytoplasm, turning on a downstream pathway that activates other transcription cofactors that interact with phosphorylated CREB to induce gene transcription. PMID- 11886857 TI - Heterogeneous nuclear ribonucleoprotein A3, a novel RNA trafficking response element-binding protein. AB - The cis-acting response element, A2RE, which is sufficient for cytoplasmic mRNA trafficking in oligodendrocytes, binds a small group of rat brain proteins. Predominant among these is heterogeneous nuclear ribonucleoprotein (hnRNP) A2, a trans-acting factor for cytoplasmic trafficking of RNAs bearing A2RE-like sequences. We have now identified the other A2RE-binding proteins as hnRNP A1/A1(B), hnRNP B1, and four isoforms of hnRNP A3. The rat and human hnRNP A3 cDNAs have been sequenced, revealing the existence of alternatively spliced mRNAs. In Western blotting, 38-, 39-, 41-, and 41.5-kDa components were all recognized by antibodies against a peptide in the glycine-rich region of hnRNP A3, but only the 41- and 41.5-kDa bands bound antibodies to a 15-residue N terminal peptide encoded by an alternatively spliced part of exon 1. The identities of these four proteins were verified by Edman sequencing and mass spectral analysis of tryptic fragments generated from electrophoretically separated bands. Sequence-specific binding of bacterially expressed hnRNP A3 to A2RE has been demonstrated by biosensor and UV cross-linking electrophoretic mobility shift assays. Mutational analysis and confocal microscopy data support the hypothesis that the hnRNP A3 isoforms have a role in cytoplasmic trafficking of RNA. PMID- 11886858 TI - The serine/threonine kinase Cmk2 is required for oxidative stress response in fission yeast. AB - Cmk2, a fission yeast Ser/Thr protein kinase homologous to mammalian calmodulin kinases, is essential for oxidative stress response. Cells lacking cmk2 gene were specifically sensitive to oxidative stress conditions. Upon stress, Cmk2 was phosphorylated in vivo, and this phosphorylation was dependent on the stress activated MAPK Sty1/Spc1. Co-precipitation assays demonstrated that Cmk2 binds Sty1. Furthermore, in vivo or in vitro activated Sty1 was able to phosphorylate Cmk2, and the phosphorylation occurred at the C-terminal regulatory domain at Thr 411. Cell lethality caused by overexpression of Wis1 MAPK kinase was abolished by deletion of cmk2 or by mutation of Thr-411 of Cmk2. Taken together, our data suggest that Cmk2 acts downstream of Sty1 and is an essential kinase for oxidative stress responses. PMID- 11886859 TI - Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis. AB - The chondrogenesis process requires the ordered proliferation and differentiation of chondrocytes. Insulin-like growth factor-binding protein (IGFBP)-3, well characterized as the carrier of insulin-like growth factor (IGF), has been reported to have intrinsic bioactivity that is independent of IGF binding. The mechanisms involved in this IGF-independent action are still unclear. Using the RCJ3.1C5.18 chondrogenic cells, which in culture progresses from undifferentiated to terminally differentiated chondrocytes, we have shown previously that IGFBP-3 has an IGF-independent, antiproliferative effect in undifferentiated and early differentiated but not in terminally differentiated chondrocytes. In the present study, cDNA microarray analysis was used to screen for genes: 1) that were regulated by IGFBP-3 in early but not in terminally differentiated chondrocytes; 2) that were regulated specifically by IGFBP-3, but not by IGF-I; and 3) whose regulation was abolished by coincubation of IGFBP-3 with IGF-I. Signal transducer and activator of transcription (STAT)-1 was the gene that, fulfilling the screening criteria, exhibited the greatest up-regulation by IGFBP-3 (>40-fold). STAT-1 gene up-regulation was confirmed by Northern analysis of cells treated with IGFBP-3 or transfected with an IGFBP-3 expression vector. Remarkably, similar results were obtained when cells were transfected with an IGFBP-3 mutant unable to bind IGFs, definitively demonstrating the IGF-independent action of IGFBP-3. Consistent with the up-regulation of STAT-1 mRNA, IGFBP-3 also increased STAT-1 protein expression. Furthermore, both IGFBP-3 and the IGFBP-3 mutant induced STAT-1 phosphorylation and its nuclear localization. An antisense STAT-1 oligonucleotide abolished the IGF-independent cell apoptosis induced by IGFBP-3. We have demonstrated that STAT-1 is a major intracellular signaling and transcriptional target of the IGF-independent apoptotic effect of IGFBP-3 in chondrogenesis. PMID- 11886860 TI - DNA polymerase lambda from calf thymus preferentially replicates damaged DNA. AB - A new gene (POLL), has been identified encoding the novel DNA polymerase lambda and mapped to mouse chromosome 19 and at human chromosome 10. DNA polymerase lambda contains all the critical residues involved in DNA binding, nucleotide binding, nucleotide selection, and catalysis of DNA polymerization and has been assigned to family X based on sequence homology with polymerase beta, lambda, mu, and terminal deoxynucleotidyltransferase. Here we describe a purification of DNA polymerase lambda from calf thymus that preferentially can replicate damaged DNA. By testing polymerase activity on non-damaged and damaged DNA, DNA polymerase lambda was purified trough five chromatographic steps to near homogeneity and identified as a 67-kDa polypeptide that cross-reacted with monoclonal antibodies against DNA polymerase beta and polyclonal antibodies against DNA polymerase lambda. DNA polymerase lambda had no detectable nuclease activities and, in contrast to DNA polymerase beta, was aphidicolin-sensitive. DNA polymerase lambda was a 6-fold more accurate enzyme in an M13mp2 forward mutation assay and 5-fold more accurate in an M13mp2T90 reversion system than human recombinant DNA polymerase beta. The biochemical properties of the calf thymus DNA polymerase lambda, described here for the first time, are discussed in relationship to the proposed role for this DNA polymerase in vivo. PMID- 11886861 TI - Modulation of TASK-1 (Kcnk3) and TASK-3 (Kcnk9) potassium channels: volatile anesthetics and neurotransmitters share a molecular site of action. AB - TASK-1 and TASK-3, members of the two-pore-domain channel family, are widely expressed leak potassium channels responsible for maintenance of cell membrane potential and input resistance. They are sites of action for a variety of modulatory agents, including volatile anesthetics and neurotransmitters/hormones, the latter acting via mechanisms that have remained elusive. To clarify these mechanisms, we generated mutant channels and found that alterations disrupting anesthetic (halothane) activation of these channels also disrupted transmitter (thyrotropin-releasing hormone, TRH) inhibition and did so to a similar degree. For both TASK-1 and TASK-3, mutations (substitutions with corresponding residues from TREK-1) in a six-residue sequence at the beginning of the cytoplasmic C terminus virtually abolished both anesthetic activation and transmitter inhibition. The only sequence motif identified with a classical signaling mechanism in this region is a potential phosphorylation site; however, mutation of this site failed to disrupt modulation. TASK-1 and TASK-3 differed insofar as a large portion of the C terminus was necessary for the full effects of halothane and TRH on TASK-3 but not on TASK-1. Finally, tandem-linked TASK-1/TASK-3 heterodimeric channels were fully modulated by anesthetic and transmitter, and introduction of the identified mutations either into the TASK-1 or the TASK-3 portion of the channel was sufficient to disrupt both effects. Thus, both anesthetic activation and transmitter inhibition of these channels require a region at the interface between the final transmembrane domain and the cytoplasmic C terminus that has not been associated previously with receptor signal transduction. Our results also indicate a close molecular relationship between these two forms of modulation, one endogenous and the other clinically applied. PMID- 11886862 TI - Novel mechanism of regulation of Rac activity and lamellipodia formation by RET tyrosine kinase. AB - Rac activation in neuronal cells plays an important role in lamellipodia formation that is a critical event for neuritogenesis. It is well known that the Rac activity is regulated via activation of phosphatidylinositol 3-kinase (PI3K) by a variety of receptor tyrosine kinases. Here we show that increased serine phosphorylation on RET receptor tyrosine kinase following cAMP elevation promotes lamellipodia formation of neuronal cells induced by glial cell line-derived neurotrophic factor (GDNF). We identified serine 696 in RET as a putative phosphorylation site by protein kinase A and found that mutation of this serine almost completely inhibited lamellipodia formation by GDNF without affecting activation of the PI3K/AKT signaling pathway. Mutation of tyrosine 1062 in RET, whose phosphorylation is crucial for activation of PI3K, also inhibited lamellipodia formation by GDNF. Inhibition of lamellipodia formation by mutation of either serine 696 or tyrosine 1062 was associated with decrease of the Rac1 guanine nucleotide exchange factor (GEF) activity, suggesting that this activity is regulated by two different signaling pathways via serine 696 and tyrosine 1062 in RET. Moreover, in the presence of serine 696 mutation, lamellipodia formation was rescued by replacing tyrosine 687 with phenylalanine. These findings propose a novel mechanism that receptor tyrosine kinase modulates actin dynamics in neuronal cells via its cAMP-dependent phosphorylation. PMID- 11886863 TI - Nuclear injection of anti-pigpen antibodies inhibits endothelial cell division. AB - Endothelial cell proliferation is required for angiogenesis in both embryonic and adult tissues. In rat brain tumors, it has recently been shown that the nuclear protein pigpen is expressed selectively in endothelial cells of developing microvasculature but not in the established peritumoral vessels (Blank, M., Weinschenk, T., Priemer, M., and Schluesener, H. (2001) J. Biol. Chem. 276, 16464 16468). This finding suggests that pigpen may be important for promoting the undifferentiated, or "angiogenic" endothelial cell phenotype. Our studies show that pigpen protein and mRNA are expressed in actively dividing endothelial cells and down-regulated as they become confluent. Protein distribution is regulated in a cell cycle-dependent manner. We conclude that this expression pattern is important for and not simply ancillary to proliferation because nuclear microinjection of anti-pigpen Fab fragments inhibited endothelial cell division. Moreover, expression of the proliferating cell marker Ki67 was inhibited in antibody-injected cells. The absence of Ki67 suggests exit from rather than arrest within (for example, at the G(1)/S interface) the cell cycle. Together with earlier observations on the structure and expression of this molecule, our data support the hypothesis that pigpen helps regulate endothelial cell differentiation state. PMID- 11886864 TI - Prp43 is an essential RNA-dependent ATPase required for release of lariat-intron from the spliceosome. AB - The essential Saccharomyces cerevisiae PRP43 gene encodes a 767-amino acid protein of the DEXH-box family. Prp43 has been implicated in spliceosome disassembly (Arenas, J. E., and Abelson, J. N. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 11798-11802). Here we show that purified recombinant Prp43 is an RNA dependent ATPase. Alanine mutations at conserved residues within motifs I ((119)GSGKT(123)), II ((215)DEAH(218)) and VI ((423)QRAGRAGR(430)) that diminished ATPase activity in vitro were lethal in vivo, indicating that ATP hydrolysis is necessary for the biological function of Prp43. Overexpression of lethal, ATPase-defective mutants in a wild-type strain resulted in dominant negative growth inhibition. The ATPase-defective mutant T123A interfered in trans with the in vitro splicing function of wild-type Prp43. T123A did not affect the chemical steps of splicing or the release of mature mRNA from the spliceosome, but it blocked the release of the excised lariat-intron from the spliceosome. We show that the lariat-intron is not accessible to debranching by purified Dbr1 when it is held in the T123A-arrested splicing complex. Our results define a new ATP-dependent step of splicing that is catalyzed by Prp43. PMID- 11886865 TI - Exon skipping in cardiac troponin T of turkeys with inherited dilated cardiomyopathy. AB - Troponin T is a central component of the thin filament-associated troponin tropomyosin system and plays an essential role in the Ca(2+) regulation of striated muscle contraction. The importance of the structure and function of troponin T is evident in the regulated isoform expression during development and the point mutations resulting in familial hypertrophic and dilated cardiomyopathies. We report here that turkeys with inherited dilated cardiomyopathy and heart failure express an unusual low molecular weight cardiac troponin T missing 11 amino acids due to the splice out of the normally conserved exon 8-encoded segment. The deletion of a 9-bp segment from intron 7 of the turkey cardiac troponin T gene may be responsible for the weakened splicing of the downstream exon 8 during mRNA processing. The exclusion of the exon 8-encoded segment results in conformational changes in cardiac troponin T, an altered binding affinity for troponin I and tropomyosin, and an increased calcium sensitivity of the actomyosin ATPase. Expression of the exon 8-deleted cardiac troponin T prior to the development of cardiomyopathy in turkeys indicates a novel RNA splicing disease and provides evidence for the role of troponin T structure-function variation in myocardial pathogenesis and heart failure. PMID- 11886866 TI - 31P NMR detection of subcellular creatine kinase fluxes in the perfused rat heart: contractility modifies energy transfer pathways. AB - The subcellular fluxes of exchange of ATP and phosphocreatine (PCr) between mitochondria, cytosol, and ATPases were assessed by (31)P NMR spectroscopy to investigate the pathways of energy transfer in a steady state beating heart. Using a combined analysis of four protocols of inversion magnetization transfer associated with biochemical data, three different creatine kinase (CK) activities were resolved in the rat heart perfused in isovolumic control conditions: (i) a cytosolic CK functioning at equilibrium (forward, F(f) = PCr --> ATP, and reverse flux, F(r) = ATP --> PCr = 3.3 mm.s(-1)), (ii) a CK localized in the vicinity of ATPases (MM-CK bound isoform) favoring ATP synthesis (F(f) = 1.7 x F(r)), and (iii) a mitochondrial CK displaced toward PCr synthesis (F(f) = 0.3 and F(r) = 2.6 mm.s(-1)). This study thus provides the first experimental evidence that the energy is carried from mitochondria to ATPases by PCr (i.e. CK shuttle) in the whole heart. In contrast, a single CK functioning at equilibrium was sufficient to describe the data when ATP synthesis was partly inhibited by cyanide (0.15 mm). In this case, ATP was directly transferred from mitochondria to cytosol suggesting that cardiac activity modified energy transfer pathways. Bioenergetic implications of the localization and activity of enzymes within myocardial cells are discussed. PMID- 11886867 TI - Cytochrome c sorption-desorption effects on the external NADH oxidation by mitochondria: experimental and computational study. AB - The rupture of the outer mitochondrial membrane is known to be critical for cell death, but the mechanism, specifically its redox-signaling aspects, still needs to be studied in more detail. In this work, the external NADH oxidation by rat liver mitochondria was studied under the outer membrane rupture induced by the mitochondria hypotonic treatment or the inner membrane permeability transition. The saturation of the oxidation rate was observed as a function of mitochondrial protein concentration. This effect was shown to result from cytochrome c binding to the mitochondrial membranes. At a relatively high concentration of mitochondria, the oxidation rate was strongly activated by 4 mm Mg(2+) due to cytochrome c desorption from the membranes. A minimal kinetic model was developed to explain the main phenomena of the external NADH oxidation modulated by cytochrome c and Mg(2+) in mitochondria with the ruptured outer membrane. The computational behavior of the model closely agreed with the experimental data. We suggest that the redox state of the released cytochrome c, considered by other authors to be important for apoptosis, may strongly depend on its oxidation by the fraction of mitochondria with the ruptured outer membrane and on the cytoplasmic cytochrome c reductase activity. PMID- 11886868 TI - Proteomic analysis of protein phosphorylations in heat shock response and thermotolerance. AB - Heat shock (HS) induces a wide variety of biological processes, including inhibition of protein synthesis, elevated expression of heat shock proteins, induction of thermotolerance, and apoptotic cell death in a dose-dependent manner. We compared phosphorylated proteins in heat-shocked and thermotolerant cells using proteome analysis. After HS treatment of control RIF-1 and their thermotolerant derivatives, TR-RIF-1 cells, cellular proteins were separated by two-dimensional gel electrophoresis and the phosphorylated proteins were detected with the anti-phosphotyrosine antibodies. We found that 93 proteins showed significant changes in phosphorylation between control and thermotolerant cells as a function of recovery time after HS; we identified 81 of these proteins with peptide mass fingerprinting using MALDI-TOF MS after in-gel trypsin digestion. These phosphorylated proteins exhibit various cellular functions, including chaperones, ion channels, signaling molecules, in transcription and translation processes, in amino acid biosynthesis, oxidoreduction, energy metabolism, and cell motility or structure, suggesting that HS turns on the various signaling pathways by activating protein-tyrosine kinases (PTKs). Of these, 20 proteins were previously identified phosphorylated proteins and 64 were newly identified. These proteins can be grouped into three families: 1) proteins highly phosphorylated in TR-RIF-1 cells at basal level and phosphorylated more significantly by HS in RIF-1 than TR-RIF-1; 2) proteins highly phosphorylated in control RIF-1 cells at basal level and phosphorylated more easily by HS in TR-RIF 1 than in RIF-1 cells; and 3) proteins with a similar basal phosphorylation level in both RIF-1 and TR-RIF-1 cells and responding to HS similarly in both cells. Most of the phosphorylated proteins are presumably involved in HS signaling in different ways, with the first and second families of proteins influencing thermotolerance. The possible tyrosine phosphorylation sites, the possible PTKs phosphorylating these proteins, and the proteins binding to these phosphorylated sites were predicted by the Netphos, ScanProsite, and Scansite programs. These results suggest that HS can activate various PTKs and HS responses can be regulated by phosphorylations of proteins having various functions. PMID- 11886870 TI - Sequestration of epidermal growth factor receptors in non-caveolar lipid rafts inhibits ligand binding. AB - Cholesterol depletion has been shown to increase mitogen-activated protein kinase activation in response to stimulation with epidermal growth factor (EGF) (Furuchi, T., and Anderson, R. G. W. (1998) J. Biol. Chem. 273, 21099-21104). However, the underlying mechanisms are unknown. We show that cholesterol depletion increases EGF binding, whereas cholesterol loading lowers EGF binding. Based on binding analyses, we demonstrate that the observed changes in EGF binding are caused by alterations in the number of EGF receptors available for ligand binding, whereas the affinity of the receptor for EGF remains unaltered. We also show by immunofluorescence that in unstimulated cells the EGF receptor is localized in non-caveolar lipid rafts containing the ganglioside GM1 and that patching of these rafts by cholera toxin B-chain causes co-patching of EGF receptors. Experiments with solubilization in different detergents at 4 degrees C show that the association of the EGF receptor with these rafts is sensitive to Triton X-100 extraction but insensitive to extraction with another non-ionic detergent, Brij 58. Furthermore, experiments with cholesterol-depleted cells show that the association is cholesterol-dependent. We propose that non-caveolar lipid rafts function as negative regulators of EGF receptor signaling by sequestering a fraction of the EGF receptors in a state inaccessible for ligand binding. PMID- 11886869 TI - A transporter in the endoplasmic reticulum of Schizosaccharomyces pombe cells mediates zinc storage and differentially affects transition metal tolerance. AB - The cation diffusion facilitator (CDF) family represents a class of ubiquitous metal transporters. Inactivation of a CDF in Schizosaccharomyces pombe, Zhf, causes drastically different effects on the tolerance toward various metals. A deletion mutant is Zn(2+)/Co(2+)-hypersensitive yet displays significantly enhanced Cd(2+) and Ni(2+) tolerance. Accumulation of zinc, cobalt, and cadmium is reduced in mutant cells. Non-vacuolar zinc content, as measured by analytical electron microscopy, is lower in zhf(-) cells compared with wild-type cells in the presence of elevated Zn(2+) concentrations. The protective effect against cadmium toxicity is independent of the phytochelatin detoxification pathway. Phytochelatin synthase-deficient cells show extremely enhanced (about 200-fold) cadmium tolerance when zhf is disrupted. Immunogold labeling indicates endoplasmic reticulum (ER) localization of Zhf. Electron spectroscopic imaging shows that accumulation of zinc coincides with Zhf localization, demonstrating a major role of the ER for metal storage and the involvement of Zhf in cellular zinc homeostasis. Also, these observations indicate that Cd(2+) ions exert their toxic effects on cellular metabolism in the ER rather than in the cytosol. PMID- 11886871 TI - Reaction mechanism of alanine racemase from Bacillus stearothermophilus: x-ray crystallographic studies of the enzyme bound with N-(5'-phosphopyridoxyl)alanine. AB - The crystal structures of alanine racemase bound with reaction intermediate analogs, N-(5'-phosphopyridoxyl)-L-alanine (PLP-L-Ala) and N-(5' phosphopyridoxyl)-D-alanine (PLP-D-Ala), were determined at 2.0-A resolution with the crystallographic R factor of 17.2 for PLP-L-Ala and 16.9 for PLP-D-Ala complexes. They were quite similar not only to each other but also to the structure of the native pyridoxal 5'-phosphate (PLP)-form enzyme; root mean square deviations at Calpha among the three structures were less than 0.28 A. The side chains of the amino acid residues around the PLP-L-Ala and PLP-D-Ala were virtually superimposable on each other as well as on those around PLP of the native holoenzyme. The alpha-hydrogen of the alanine moiety of PLP-L-Ala was located near the OH of Tyr(265)', whereas that of PLP-D-Ala was near the NZ of Lys(39). These support the previous findings that Tyr(265)' and Lys(39) are the catalytic bases removing alpha-hydrogen from L- and D-alanine, respectively. The prerequisite for this two-base mechanism is that the alpha-proton abstracted from the substrate is transferred (directly or indirectly) between the NZ of Lys(39) and the OH of Tyr(265'); otherwise the enzyme reaction stops after a single turnover. Only the carboxylate oxygen atom of either PLP-Ala enantiomer occurred at a reasonable position that can mediate the proton transfer; neither the amino acid side chains nor the water molecules were located in the vicinity. Therefore, we propose a mechanism of alanine racemase reaction in which the substrate carboxyl group directly participates in the catalysis by mediating the proton transfer between the two catalytic bases, Lys(39) and Tyr(265)'. The results of molecular orbital calculation also support this mechanism. PMID- 11886872 TI - Unique molecular architecture of silk fibroin in the waxmoth, Galleria mellonella. AB - Proteins of silk fibers are characterized by reiterations of amino acid repeats. Physical properties of the fiber are determined by the amino acid composition, the complexity of repetitive units, and arrangement of these units into higher order arrays. Except for very short motifs of 6-10 residues, the length of repetitive units and the number of these units concatenated in higher order assemblies vary in all spider and lepidopteran silks analyzed so far. This paper describes an exceptional silk protein represented by the 500-kDa heavy chain fibroin (H-fibroin) of the waxmoth, Galleria mellonella. Its non-repetitive N terminal (175 residues) and C-terminal (60 residues) parts, the overall gene organization, and the nucleotide sequence around the TATA box show that it is homologous to the H-fibroins of other Lepidoptera. However, over 95% of the protein consists of highly ordered repetitive structures that are unmatched in other species. The repetitive region includes 11 assemblies AB(1)AB(1)AB(1)AB(2)(AB(2))AB(2) of remarkably conserved polypeptide repeats A (63 amino acid residues), B(1) (43 residues), and B(2) (18 residues). The repeats contain a high proportion of Gly (31.6%), Ala (23.8%), Ser (18.1%), and of residues with long hydrophobic side chains (16% for Leu, Ile, and Val combined). The presence of the GLGGLG and SSAASAA(AA) motifs suggests formation of pleated beta-sheets and their stacking into crystallites. Conspicuous conservation of the apolar sequence VIVI followed by DD or ED is interpreted as indicating the importance of hydrophobicity and electrostatic charge in H-fibroin cross-linking. The environment of G. mellonella larvae within bee cultures requires continuous production of silk that must be both strong and elastic. The spectacular arrangement of the repetitive H-fibroin region apparently evolved to meet these requirements. PMID- 11886873 TI - Characterization of neuropilin-1 structural features that confer binding to semaphorin 3A and vascular endothelial growth factor 165. AB - Neuropilin-1 (Npn-1) is a receptor for both semaphorin 3A (Sema3A) and vascular endothelial growth factor 165 (VEGF(165)). To understand the role Npn-1 plays as a receptor for these structurally and functionally unrelated ligands, we set out to identify structural features of Npn-1 that confer binding to Sema3A or VEGF(165). We constructed Npn-1 variants containing deletions within the "a" and "b" domains of Npn-1. More than 16 variants were expressed in COS-1 cells and tested for alkaline phosphatase-Sema3A as well as alkaline phosphatase-VEGF(165) binding. Our results indicate that each of the two Npn-1 CUB domains and the amino-terminal coagulation factor V/VIII domain (CF V/VIII) are essential for Sema3A binding, but only the amino-terminal Npn-1 CF V/VIII domain is required for binding to VEGF(165). Guided by the structure of the bovine spermadhesin CUB domain, point mutants targeting defined surfaces of the Npn-1 a1 CUB domain were generated and tested for Sema3A and VEGF(165) binding. One Npn-1 variant, Npn 1(2ABC), exhibits complete loss of Sema3A binding while retaining normal VEGF(165) binding. Moreover, co-immunoprecipitation experiments show that Npn 1(2ABC) can form a signaling complex with the VEGF(165) signaling receptor KDR/VEGFR-2. These results establish the identity of contact sites between Npn-1 and its semaphorin ligands, and they provide a foundation for understanding how Npn-1 functions as a receptor for distinct classes of ligands in vivo. PMID- 11886874 TI - Interaction of the cation-dependent mannose 6-phosphate receptor with GGA proteins. AB - The GGAs (Golgi-localizing, gamma-adaptin ear homology domain, ARF-binding) are a multidomain family of proteins implicated in protein trafficking between the Golgi and endosomes. Recent evidence has established that the cation-independent (CI) and cation-dependent (CD) mannose 6-phosphate receptors (MPRs) bind specifically to the VHS domains of the GGAs through acidic cluster-dileucine motifs at the carboxyl ends of their cytoplasmic tails. However, the CD-MPR binds the VHS domains more weakly than the CI-MPR. Alignment of the C-terminal residues of the two receptors revealed a number of non-conservative differences in the acidic cluster-dileucine motifs and the flanking residues. Mutation of these residues in the CD-MPR cytoplasmic tail to the corresponding residues in the CI MPR conferred either full binding (H63D mutant), intermediate binding (R60S), or unchanged binding (E56F/S57H) to the GGAs as determined by in vitro glutathione S transferase pull-down assays. Furthermore, the C-terminal methionine of the CD MPR, but not the C-terminal valine of the CI-MPR, inhibited GGA binding. Addition of four alanines to the C-terminal valine of the CI-MPR also severely reduced GGA binding, demonstrating the importance of the spacing of the acidic cluster dileucine motif relative to the C terminus for optimal GGA interaction. Mouse L cells stably expressing CD-MPRs with mutations that enhance GGA binding sorted cathepsin D more efficiently than wild-type CD-MPR. These studies provide an explanation for the observed differences in the relative affinities of the two MPRs for the GGA proteins. Furthermore, they indicate that the GGAs participate in lysosomal enzyme sorting mediated by the CD-MPR. PMID- 11886875 TI - Contributions of the LG modules and furin processing to laminin-2 functions. AB - The alpha2-laminin subunit contributes to basement membrane functions in muscle, nerve, and other tissues, and mutations in its gene are causes of congenital muscular dystrophy. The alpha2 G-domain modules, mutated in several of these disorders, are thought to mediate different cellular interactions. To analyze these contributions, we expressed recombinant laminin-2 (alpha(2)beta(1)gamma(1)) with LG4-5, LG1-3, and LG1-5 modular deletions. Wild-type and LG4-5 deleted laminins were isolated from medium intact and cleaved within LG3 by a furin-like convertase. Myoblasts adhered predominantly through LG1-3 while alpha dystroglycan bound to both LG1-3 and LG4-5. Recombinant laminin stimulated acetylcholine receptor (AChR) clustering; however, clustering was induced only by the proteolytic processed form, even in the absence of LG4-5. Furthermore, clustering required alpha(6)beta(1) integrin and alpha-dystroglycan binding activities available on LG1-3, acting in concert with laminin polymerization. The ability of the modified laminins to mediate basement membrane assembly was also evaluated in embryoid bodies where it was found that both LG1-3 and LG4-5, but not processing, were required. In conclusion, there is a division of labor among LG-modules in which (i) LG4-5 is required for basement membrane assembly but not for AChR clustering, and (ii) laminin-induced AChR clustering requires furin cleavage of LG3 as well as alpha-dystroglycan and alpha(6)beta(1) integrin binding. PMID- 11886876 TI - Identification and molecular characterization of two closely related G protein coupled receptors activated by prokineticins/endocrine gland vascular endothelial growth factor. AB - We previously described two mammalian secreted proteins, prokineticin 1 and prokineticin 2, that potently contract gastrointestinal smooth muscle. Prokineticin 1 has also been shown to promote angiogenesis by stimulating proliferation, migration, and fenestration of endocrine organ-derived endothelial cells. Here we report the cloning and characterization of two closely related G protein-coupled receptors as receptors for prokineticins. Expression of prokineticin receptors in heterologous systems shows that these receptors bind to and are activated by nanomolar concentrations of recombinant prokineticins. Activation of prokineticin receptors leads to mobilization of calcium, stimulation of phosphoinositide turnover, and activation of p44/p42 MAPK signaling pathways that are consistent with the effects of prokineticins on smooth muscle contraction and angiogenesis. mRNA expression analysis reveals that prokineticin receptors are expressed in gastrointestinal organs, endocrine glands, and other tissues. PMID- 11886877 TI - Crassulacean acid metabolism: plastic, fantastic. AB - The occurrence, activity and plasticity of the CAM pathway is described from an introductory viewpoint, framed by the use of the four "Phases" of CAM as comparative indicators of the interplay between environmental constraints and internal molecular and biochemical regulation. Having described a number of "rules" which seem to govern the CAM cycle and apply uniformly to most species, a number of key regulatory points can then be identified. These include temporal separation of carboxylases, based on the circadian expression of key genes and their control by metabolites. The role of a circadian oscillator and interplay between tonoplast and nuclear control are central to maintaining the CAM cycle. Control of reserve carbohydrates is often neglected, but the importance of daily partitioning (for growth and the subsequent night-time CAM activity) and use at night is shown to drive the CAM cycle. Finally, it is shown that the genotypic and phenotypic plasticity in patterns of CAM expression is mediated partly by environmental conditions and molecular signalling, but also by diffusive constraints in succulent tissues. A transformation system is now required to allow these key areas of control to be elucidated. PMID- 11886878 TI - C(4) photosynthesis: principles of CO(2) concentration and prospects for its introduction into C(3) plants. AB - C(4) photosynthesis has a number of distinct properties that enable the capture of CO(2) and its concentration in the vicinity of Rubisco, so as to reduce the oxygenase activity of Rubisco, and hence the rate of photorespiration. The aim of this review is to discuss the properties of this CO(2)-concentrating mechanism, and thus to indicate the minimum requirements of any genetically-engineered system. In particular, the Kranz leaf anatomy of C(4) photosynthesis and the division of the C(4)-cycle between two cell types involves intercellular co operation that requires modifications in regulation and transport to make C(4) photosynthesis work. Some examples of these modifications are discussed. Comparisons are made with the C(4)-type photosynthesis found in single-celled C(4)-type CO(2)-concentrating mechanisms, such as that found in the aquatic plant, Hydrilla verticillata and the single-celled C(4) system found in the terrestrial chenopod Borszczowia aralocaspica. The outcome of recent attempts to engineer C(4) enzymes into C(3) plants is discussed. PMID- 11886879 TI - Overexpression of C(4)-cycle enzymes in transgenic C(3) plants: a biotechnological approach to improve C(3)-photosynthesis. AB - The process of photorespiration diminishes the efficiency of CO(2) assimilation and yield of C(3)-crops such as wheat, rice, soybean or potato, which are important for feeding the growing world population. Photorespiration starts with the competitive inhibition of CO(2) fixation by O(2) at the active site of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and can result in a loss of up to 50% of the CO(2) fixed in ambient air. By contrast, C(4) plants, such as maize, sugar cane and Sorghum, possess a CO(2) concentrating mechanism, by which atmospheric CO(2) is bound to C(4)-carbon compounds and shuttled from the mesophyll cells where the prefixation of bicarbonate occurs via phosphoenolpyruvate carboxylase (PEPC) into the gas-tight bundle-sheath cells, where the bound carbon is released again as CO(2) and enters the Calvin cycle. However, the anatomical division into mesophyll and bundle-sheaths cells ("Kranz" anatomy) appears not to be a prerequisite for the operation of a CO(2) concentrating mechanism. Submerged aquatic macrophytes, for instance, can induce a C(4)-like CO(2) concentrating mechanism in only one cell type when CO(2) becomes limiting. A single cell C(4)-mechanism has also been reported recently for a terrestrial chenopod. For over 10 years researchers in laboratories around the world have attempted to improve photosynthesis and crop yield by introducing a single cell C(4)-cycle in C(3) plants by a transgenic approach. In the meantime, there has been substantial progress in overexpressing the key enzymes of the C(4) cycle in rice, potato, and tobacco. In this review there will be a focus on biochemical and physiological consequences of the overexpression of C(4) cycle genes in C(3) plants. Bearing in mind that C(4)-cycle enzymes are also present in C(3) plants, the pitfalls encountered when C(3) metabolism is perturbed by the overexpression of individual C(4) genes will also be discussed. PMID- 11886880 TI - Variation in the k(cat) of Rubisco in C(3) and C(4) plants and some implications for photosynthetic performance at high and low temperature. AB - The capacity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to consume RuBP is a major limitation on the rate of net CO(2) assimilation (A) in C(3) and C(4) plants. The pattern of Rubisco limitation differs between the two photosynthetic types, as shown by comparisons of temperature and CO(2) responses of A and Rubisco activity from C(3) and C(4) species. In C(3) species, Rubisco capacity is the primary limitation on A at light saturation and CO(2) concentrations below the current atmospheric value of 37 Pa, particularly near the temperature optimum. Below 20 degrees C, C(3) photosynthesis at 37 and 68 Pa is often limited by the capacity to regenerate phosphate for photophosphorylation. In C(4) plants, the Rubisco capacity is equivalent to A below 18 degrees C, but exceeds the photosynthetic capacity above 25 degrees C, indicating that Rubisco is an important limitation at cool but not warm temperatures. A comparison of the catalytic efficiency of Rubisco (k(cat) in mol CO(2) mol(-1) Rubisco active sites s(-1)) from 17 C(3) and C(4) plants showed that Rubisco from C(4) species, and C(3) species originating in cool environments, had higher k(cat) than Rubisco from C(3) species originating in warm environments. This indicates that Rubisco evolved to improve performance in the environment that plants normally experience. In C(4) plants, and C(3) species from cool environments, Rubisco often operates near CO(2) saturation, so that increases in k(cat) would enhance A. In warm-habitat C(4) species, Rubisco often operates at CO(2) concentrations below the K(m) for CO(2). Because k(cat) and K(m) vary proportionally, the low k(cat) indicates that Rubisco has been modified in a manner that reduces K(m) and thus increases the affinity for CO(2) in C(3) species from warm climates. PMID- 11886881 TI - Local expression of the ipt gene in transgenic tobacco (Nicotiana tabacum L. cv. SR1) axillary buds establishes a role for cytokinins in tuberization and sink formation. AB - The developmental characteristics of a transgenic tobacco line (BIK62) expressing the ipt cytokinin-biosynthetic gene under the control of a tagged promoter were analysed. In situ hybridization and cytokinin immunocytochemistry revealed that the ipt gene was mainly expressed in the axillary buds after the floral transition. The ipt-expressing axillary buds presented morphological alterations such as short and narrow scale-leaflets, and swollen internodes filled with starch grains, giving rise to short and tuberized lateral branches. In addition, the modification of the endogenous cytokinin balance in the axillary meristems resulted in a fast rate of leaf initiation and cytokinins accumulated mostly in the lateral zones of the reactivated axillary meristems, suggesting a role in leaf organogenesis. Cell cycle analysis revealed that the reactivated axillary meristems were characterized by predominant S+G2 nuclei. Terminal internodes displayed low levels of hexose and sucrose concomitant with starch accumulation. Extracellular invertases (EC 3.1.26) were also present in higher amounts in the tuberizing internodes compared to the axillary buds of wild-type tobacco. These results underline the role of cytokinins in cell cycle regulation and in the creation of a sink--source effect. They also provide new information about cytokinin involvement in the process of tuberization and their overproduction in axillary buds giving rise to tuberized lateral branches in a naturally non tuberizing species. PMID- 11886883 TI - High expression of putative aquaporin genes in cells with transporting and nutritive functions during seed development in Norway spruce (Picea abies). AB - Aquaporins mediate the bidirectional passage of water over membranes and are present in tonoplasts (TIPs) and in plasma membranes (PIPs) of plant cells. Knowing their expression in different tissues is valuable when assessing their contribution to plant water relations. A TIP-gene has been cloned from developing female gametophytes of Picea abies, a conifer displaying an embryology different from the angiosperms. Probes were made from conserved regions of the TIP gene and used for in situ hybridization to examine the gene expression pattern in developing female reproductive structures. Early during development high transcript expression was found in the spongy tissue encasing the developing female gametophyte, in cells of the future seed coat of young ovules and in vascular tissue of the ovuliferous scale. At later stages a strong signal was seen in archegonia jacket cells surrounding egg cells and, still later, at the time of storage protein accumulation, in storage parenchyma cells of the gametophyte as well. These aquaporin-homologues probably participate in regulating water balance in the cells although they could also be permeable to other molecules than water. PMID- 11886882 TI - Use of aphid stylectomy and RT-PCR for the detection of transporter mRNAs in sieve elements. AB - Unmodified samples of barley (Hordeum vulgare) sieve tube sap have been obtained by severing the stylets (stylectomy) of feeding aphids and collecting the exuding liquid. Primers were designed to direct the amplification of a series of specific cDNAs encoding barley proteins selected because of their significance in sieve tube function. mRNA encoding the H(+)/sucrose co-transporter SUT1, a putative aquaporin and the H(+)/ATPase PPA1 were detected in sieve tube sap. These mRNA species appear to be present at very low concentrations. mRNA encoding the potassium transporter HAK1 could not be detected. The results strongly suggest that some mRNA species are imported into sieve elements, which are enucleate, from neighbouring companion cells. PMID- 11886884 TI - Identification of programmed cell death in situ in individual plant cells in vivo using a chromosome preparation technique. AB - A simple procedure, which combines a chromosome preparation technique with an in situ labelling technique modified from fluorescence in situ hybridization (FISH), has been developed for in situ detection of plant programmed cell death (PCD) at the single-cell level. After exposure of chromosomes and nuclei on slides by enzymolysis, Klenow or TdT was used to incorporate Bio-dUTP or fluorescein-dUTP at sites of DNA breaks. After Klenow-mediated labelling, the signals were amplified by a cascade of antigen-antibody reaction according to the detection system of FISH. This method enables in situ detection of plant PCD in vivo morphologically and biochemically at the chromosome, nuclear and DNA levels without cell culture and histological sectioning. This technique permits labelling of DNA breaks with high sensitivity due to increased chromosome and nucleus exposure to the labelling solutions, as well as due to the immunological amplification of the signals. Moreover, the changes in the cells were easier to be observed because the spatial obstacle of the cell wall and its autofluorescence were eliminated. It is potentially useful for in situ detection of PCD in plant root meristematic cells triggered by various environmental abiotic factors. It is proposed that the root tip is a versatile in vivo system for studying PCD induced by environmental abiotic factors. PMID- 11886885 TI - Plant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules. AB - Mitochondrion movement and positioning was studied in elongating cultured cells of tobacco (Nicotiana tabacum L.), containing mitochondria-localized green fluorescent protein. In these cells mitochondria are either actively moving in strands of cytoplasm transversing or bordering the vacuole, or immobile positioned in the cortical layer of cytoplasm. Depletion of the cell's ATP stock with the uncoupling agent DNP shows that the movement is much more energy demanding than the positioning. The active movement is F-actin based. It is inhibited by the actin filament disrupting drug latrunculin B, the myosin ATPase inhibitor 2,3-butanedione 2-monoxime and the sulphydryl-modifying agent N ethylmaleimide. The microtubule disrupting drug oryzalin did not affect the movement of mitochondria itself, but it slightly stimulated the recruitment of cytoplasmic strands, along which mitochondria travel. The immobile mitochondria are often positioned along parallel lines, transverse or oblique to the cell axis, in the cortical cytoplasm of elongated cells. This positioning is mainly microtubule based. After complete disruption of the F-actin, the mitochondria parked themselves into conspicuous parallel arrays transverse or oblique to the cell axis or clustered around chloroplasts and around patches and strands of endoplasmic reticulum. Oryzalin inhibited all positioning of the mitochondria in parallel arrays. PMID- 11886886 TI - Endoreduplication is not inhibited but induced by aphidicolin in cultured cells of tobacco. AB - Endoreduplication is a common process in plants that allows cells to increase their DNA content. In the tobacco cell cultures studied in this work it can be induced by simple hormone deprivation. Mesophyll protoplast-derived cells cultured in the presence of NAA (auxin) and BAP (cytokinin) keep on dividing, while elongation and concomitant DNA endoreduplication are induced and maintained in a medium containing only NAA. If aphidicolin is given to the two types of culture, no effect is observed on elongating, endoreduplicating cells. However, the cells programmed for division switch to elongation and DNA endoreduplication. Thus aphidicolin, an inhibitor of the replicative DNA polymerases, alpha and delta, does not inhibit endoreduplication, and furthermore actually induces it when the mitotic cell cycle is blocked. DNA duplication and cell growth can only be completely blocked if ddTTP, an inhibitor of DNA polymerase-beta, is given together with aphidicolin. This result implies that an aphidicolin-resistant DNA polymerase, such as the repair-associated DNA polymerase-beta, can mediate DNA synthesis during endoreduplication and can substitute for polymerases-alpha and delta when the latter are inhibited. Similar results are obtained in cultures of the BY-2 cell line by withdrawing auxins from the culture medium. In this cell line endoreduplication is induced only in a small proportion of the cells. A greater proportion of the cells are blocked in the G(2) phase of the cell cycle. PMID- 11886887 TI - Diamine oxidase is involved in H(2)O(2) production in the chalazal cells during barley grain filling. AB - The localization and activities of diamine oxidase (DAO, EC 1.4.3.6) and polyamine oxidase (PAO, EC 1.4.3.4) together with polyamine levels have been investigated in developing grains of barley (Hordeum vulgare L.). DAO (pH 7.5) is present mainly in vascular tissue and its neighbouring cells, namely chalazal cells and nucellar projection, while PAO (pH 6.0) is mainly localized in the chlorenchymatous cells of the crease and at the base of the vascular tissue. Activities of both these enzymes appear to be independently-regulated, as DAO activity increased steadily throughout grain development while PAO activity was higher during the early stages of grain filling, declined thereafter and again increased towards maturity. The maximum activities of DAO coincided with the maximum content of putrescine while the levels of PAO did not seem to be directly correlated with spermidine or spermine contents. Isoelectric focusing (IEF) of DAO and PAO activities revealed the presence of bands at 30 and 45 DPA. The possible involvement of DAO and PAO in the supply of H(2)O(2) to peroxidase catalysed reactions in the chalazal cells during grain filling is discussed. PMID- 11886888 TI - Positional effect of cell inactivation on root gravitropism using heavy-ion microbeams. AB - When primary root apical tissues of Arabidopsis thaliana were irradiated by heavy ion microbeams with 120 microm diameter, strong inhibition of root elongation and curvature were observed at the root tip. Irradiation of the cells that become the lower part of the root cap after gravistimulation showed strong inhibition of root curvature, whereas irradiation of the cells that become the upper part of the root cap after gravistimulation did not show severe damage in either root curvature or root growth. Further analysis using smaller area microbeams with 40 microm diameter indicated that the greatest inhibition of curvature occurred at the root tip and the next greatest inhibition occurred in the cells in the lower part of the root cap. These results indicate not only that the root tip and columella cells are the most sensitive sites for root gravity, but also that signalling of root gravity would go through the lower part of the cap cells after perception. PMID- 11886889 TI - Spatio-temporal dynamics of expansion growth in roots: automatic quantification of diurnal course and temperature response by digital image sequence processing. AB - A newly developed technique based on image sequence analysis allows automatic and precise quantification of the dynamics of the growth velocity of the root tip, the distribution of expansion growth rates along the entire growth zone and the oscillation frequencies of the root tip during growth without the need of artificial landmarks. These three major parameters characterizing expansion growth of primary roots can be analysed over several days with high spatial (20 microm) and temporal resolution (several minutes) as the camera follows the growing root by an image-controlled root tracking device. In combination with a rhizotron set up for hydroponic plant cultivation the impact of rapid changes of environmental factors can be assessed. First applications of this new system proved the absence of diurnal variation of root growth in Zea mays under constant temperature conditions. The distribution profile of relative elemental growth rate (REGR) showed two maxima under constant and varying growth conditions. Lateral oscillatory movements of growing root tips were present even under constant environmental conditions. Dynamic changes in velocity- and REGR distribution within 1 h could be quantified after a step change in temperature from 21 degrees C to 26 degrees C. Most prominent growth responses were found in the zone of maximal root elongation. PMID- 11886890 TI - Engineering for drought avoidance: expression of maize NADP-malic enzyme in tobacco results in altered stomatal function. AB - Water is a principal limitation to agricultural production during drought and in arid regions of the world. Mechanisms that plants use to cope with drought can be grouped into two different strategies: drought tolerance and drought avoidance. Previous efforts toward engineering plants for improved performance during drought have focused on drought tolerance, the ability to adjust to dry conditions. This report addresses the engineering of a drought-avoidance phenotype, which allows for the conservation of water during plant growth. The majority of water lost from plants occurs through stomata. When stomata are open, potassium, chloride and/or malate are present at high concentrations in guard cells. The accumulation of large numbers of ions during stomatal opening increases the turgor pressure of the guard cells, which results in increased pore size. Expression of a single gene from maize, NADP-malic enzyme (ME), which converts malate and NADP to pyruvate, NADPH, and CO(2), resulted in altered stomatal behaviour and water relations in tobacco. The ME-transformed plants had decreased stomatal conductance and gained more fresh mass per unit water consumed than did the wild type, but they were similar to the wild type in their growth and rate of development. Providing chloride via the transpiration stream partially reversed the effects of ME expression on stomatal aperture size, which is consistent with the interpretation that expression of ME altered malate metabolism in guard cells. These results suggest a role for malic enzyme in the mechanism of stomatal closure, as well as a potential mechanism for genetically altering plant water use. PMID- 11886891 TI - Regulation of pectic polysaccharide domains in relation to cell development and cell properties in the pea testa. AB - The occurrence of pectic polysaccharide epitopes in cells and tissues of the pea testa during late stages of seed development have been examined in relation to anatomy and cell properties. Homogalacturonan, in a highly methyl-esterified form, was present throughout late development in all pea testa cell walls, including the thickened cell walls of the outer macrosclereid layer. Two epitopes, characteristic of the side-chains of the rhamnogalacturonan-I domain of pectic polysaccharides, occurred in restricted and separate cell layers of the pea testa. A (1-->4)-beta-D-galactan epitope was restricted to regions of the outer cell wall of the testa and to inner regions of the macrosclereid layer at 20 DAA and was absent from the osteosclereid and parenchyma cell walls. By 25 DAA the (1-->4)-beta-D-galactan epitope occurred only in the outer epidermal cell walls. A (1-->5)-alpha-L-arabinan epitope was also dependent on the developmental stage of the seed and was found with greatest abundance in the walls of the inner parenchyma cells. Cell separation studies indicated that, although calcium cross links were involved in the maintenance of the link between the macrosclereid layer and proximal cell layers, most cell-to-cell adhesion in the testa was not due to calcium- or ester-based bonds. PMID- 11886892 TI - A model co-ordinating the elongation of all leaves of a sorghum cultivar was applied to both Mediterranean and Sahelian conditions. AB - Sorghum leaf development was analysed at plant level by analysing the time-course of elongation and identifying the beginning and end of the elongation phases of each leaf blade. This was done with destructive and non-destructive measurements in 14 experiments carried out during several growing periods in Southern France and Sahelian Africa. Elongation of each blade was characterized by the succession of a nearly exponential phase and a linear phase. For a given blade and provided that time was expressed in thermal units, initiation, beginning and end of the linear phase, and time-courses of elongation rate were strikingly similar in all experiments, except in environments with a maximum air temperature close to 40 degrees C and a maximum vapour pressure deficit close to 6 kPa. The relative elongation rate during the exponential phase declined with leaf number from 0.08 to 0.02 degrees Cd(-1), while the duration of this phase increased from 140 to 320 degrees Cd. By contrast, the absolute elongation rate during the linear phase was nearly constant from leaf 8 onwards. This phase was shorter than the exponential phase regardless of leaf position, but accounted for the largest part of blade length. A strict pattern of leaf development was observed at the whole plant level, whereby dates of elongation events and leaf and ligule appearance, represented on a thermal time scale, were linearly related to phytomer number. This pattern exhibited a simultaneous elongation cessation of the last-formed leaves and a mismatch between real and apparent (from leaf to ligule appearance) elongation duration. PMID- 11886894 TI - Impact of elevated CO(2) and O(3) on gas exchange parameters and epidermal characteristics in potato (Solanum tuberosum L.). AB - Potato plants (Solanum tuberosum L. cv. Bintje) were grown in open-top chambers (OTCs) under three CO(2) levels (ambient and 24 h d(-1) seasonal mean concentrations of 550 and 680 micromol mol(-1)) and two O(3) levels (ambient and a seasonal mean 8 h d(-1) concentration of 50 nmol mol(-1)). The objectives were to determine the effects of season-long exposure to these key climate change gases on gas exchange, leaf thickness and epidermal characteristics. The experimental design also provided an ideal opportunity to examine within-leaf variation in epidermal characteristics at the whole-leaf level. Stomatal and epidermal cell density and stomatal index were measured at specific locations on the youngest fully expanded leaf (centre of lamina, mid-way between tip and base) and representative whole leaves from each treatment. Effects on leaf conductance, assimilation rate and instantaneous transpiration efficiency were determined by infrared gas analysis, while anatomical characteristics were examined using a combination of leaf impressions and thin sections. Exposure to elevated CO(2) or O(3) generally increased leaf thickness, leaf area, stomatal density, and assimilation rate, but reduced leaf conductance. The irregular stomatal distribution within leaves resulted from a combination of uneven differentiation and expansion of the epidermal cells. The results are discussed with reference to sampling protocols and the need to account for within-leaf variation when examining the impact of climate change or other environmental factors on epidermal characteristics. PMID- 11886893 TI - IDI7, a new iron-regulated ABC transporter from barley roots, localizes to the tonoplast. AB - A new Fe-deficiency-induced cDNA, IDI7, was isolated from the roots of Fe deficient barley (Hordeum vulgare L. cv. Ehimehadaka no. 1). The transcript levels of IDI7 in roots strongly correlated with iron nutritional status, and induction by Fe-deficiency was restricted to roots. Excess treatment with heavy metal ions, such as copper, manganese, and zinc, did not cause obvious IDI7 induction in either leaves or roots. IDI7 encodes a 644 amino acid protein, and has features typical of ATP-binding cassette (ABC) transporters. Phylogenetic analysis revealed that IDI7 is closely related to the half-type ABC protein subfamily, which includes mammalian transporters associated with antigen processing (TAPs). A transiently expressed fusion protein of IDI7 to green fluorescent protein (GFP) was localized to tonoplasts in suspension-cultured tobacco (Nicotiana tabacum L.) cells. IDI7 and its orthologues are thought to comprise a new class of ABC transporters, located in the tonoplasts of higher plants. A possible Fe-deficiency adaptation role for IDI7 in barley root cells, involving transport across the tonoplast, is proposed. PMID- 11886895 TI - Methyl jasmonate alters polyamine metabolism and induces systemic protection against powdery mildew infection in barley seedlings. AB - Treatment of the first leaves of barley (Hordeum vulgare L. cv. Golden Promise) seedlings with methyl jasmonate (MJ) led to small, but significant increases in levels of free putrescine and spermine 1 d later and to significant increases in levels of free putrescine, spermidine and spermine by 4 d following treatment. MJ treated first leaves also exhibited significant increases in the amounts of soluble conjugates of putrescine and spermidine 1, 2 and 4 d after treatment. In second leaves of plants where the first leaves had been treated with MJ, no significant changes in levels of free polyamines were observed, but significant increases in levels of soluble conjugates of putrescine and spermidine were detected. These changes were accompanied by increased activities of soluble ornithine decarboxylase (ODC), soluble and particulate arginine decarboxylase (ADC), and S-adenosylmethionine decarboxylase (AdoMetDC), in first and second leaves following treatment of the first leaves with MJ. Activities of soluble and particulate diamine oxidase (DAO) were also higher in first and second leaves following treatment of the first leaves with MJ. Treatment of the first leaves with MJ led to a significant reduction in powdery mildew (Blumeria graminis f. sp. hordei) infection on the second leaves and also resulted in significant increases in activities of the plant defence-related enzymes, phenylalanine ammonia lyase (PAL) and peroxidase. PMID- 11886896 TI - Carbon isotope composition of sugars in grapevine, an integrated indicator of vineyard water status. AB - Photosynthetic carbon isotope composition (delta(13)C) was measured on sugars in mature fruits from field-grown grapevines. Sugar delta(13)C and summer predawn leaf water potential were significantly correlated. The survey of different vineyards during four growing seasons showed that sugar delta(13)C in must at harvest varied from -20 per thousand to -26 per thousand when conditions during berry maturation varied from dry to wet. This range allows a very sensitive detection of grapevine water status under natural conditions. However, local differences due to soil capacity to supply water to grapevines are maintained, whatever the annual water balance. Leaf nitrogen content variations of field grown grapevines did not change delta(13)C values. Genetic variability of delta(13)C between 31 grapevine varieties for delta(13)C was observed. Must sugar delta(13)C can be used to characterize vineyards for their soil structural capacity to provide water to grapevines. It was concluded that isotope carbon composition in grapevine measured on sugars at harvest can be applied to compare the capacities of vineyard soils and canopy management to induce mild water stress in order to produce premium wines. PMID- 11886897 TI - Isolation of a cDNA for a nucleoside diphosphate kinase capable of phosphorylating the kinase domain of the self-incompatibility factor SRK of Brassica campestris. AB - SRK is a plant receptor kinase involved in the self-incompatibility system of Brassica species. During a cDNA screening for the phosphoproteins from a stigma expression library, a clone encoding the nucleoside diphosphate kinase III (Bc NDPK III) was obtained. After in vitro phosphorylation assays with recombinant proteins, Bc-NDPK III contained mostly phosphoserine. By contrast, the kinase domain of SRK contained phosphoserine and phosphothreonine, both of which were significantly increased by the addition of Bc-NDPK III in the presence of an SRK inhibitor KN-62. The result suggested the possible involvement of Bc-NDPK III in the signal transduction pathway through SRK. PMID- 11886898 TI - Hamy3, a novel type 100 kDa myosin from sunflower. AB - Hamy3, a novel type myosin heavy chain from sunflower is the smallest myosin described so far, with only 900 amino acid residues. One interesting finding in Hamy3 is the glycine to glutamine alteration at residue 741, which corresponds to chicken skeletal muscle myosin glycine 699 (G699). G699 is found in 125 out of 129 myosin sequences and is interpreted in terms of its role as a pivot point for motion in the myosin "lever arm hypothesis". Changes in this crucial part of myosin might indicate a role that is different from the generation of intracellular motility. PMID- 11886900 TI - Evaluation of low-energy extracorporeal shock-wave application for treatment of chronic plantar fasciitis. AB - BACKGROUND: Although the application of low-energy extracorporeal shock waves to treat musculoskeletal disorders is controversial, there has been some limited, short-term evidence of its effectiveness for the treatment of chronic plantar fasciitis. METHODS: From 1993 to 1995, a prospective, two-tailed, randomized, controlled, observer-blinded pilot trial was performed to assess whether three applications of 1000 impulses of low-energy shock waves (Group I) led to a superior clinical outcome when compared with three applications of ten impulses of low-energy shock waves (Group II) in patients with intractable plantar heel pain. The sample size was 112. The main outcome measure was patient satisfaction according to a four-step score (excellent, good, acceptable, and poor) at six months. Secondary outcome measures were patient satisfaction according to the four-step score at five years and the severity of pain on manual pressure, at night, and at rest as well as the ability to walk without pain at six months and five years. RESULTS: At six months, the rate of good and excellent outcomes according to the four-step score was significantly (47%) better (p < 0.0001) in Group I than in Group II. As assessed on a visual analog scale, the score for pain caused by manual pressure at six months had decreased to 19 points, from 77 points before treatment, in Group I, whereas in Group II the ratings before treatment and at six months were 79 and 77 points (p < 0.0001 for the difference between groups). In Group I, twenty-five of forty-nine patients were able to walk completely without pain at six months compared with zero of forty-eight patients in Group II (p < 0.0001). By five years, the difference in the rates of good and excellent outcomes according to the four-step score was only 11% in favor of Group I (p = 0.071) because of a high rate of good and excellent results from subsequent surgery in Group II; the score for pain caused by manual pressure had decreased to 9 points in Group I and to 29 points in Group II (p = 0.0006 for the difference between groups). At five years, five (13%) of thirty-eight patients in Group I had undergone an operation of the heel compared with twenty-three (58%) of forty patients in Group II (p < 0.0001). CONCLUSIONS: Three treatments with 1000 impulses of low-energy shock waves appear to be an effective therapy for plantar fasciitis and may help the patient to avoid surgery for recalcitrant heel pain. In contrast, three applications of ten impulses did not improve symptoms substantially. PMID- 11886899 TI - Mutant Cu, Zn superoxide dismutase that causes motoneuron degeneration is present in mitochondria in the CNS. AB - Mutations in Cu, Zn superoxide dismutase (SOD1) cause a fraction of amyotrophic lateral sclerosis (ALS), which involves motoneuron degeneration, paralysis, and death. An acquired activity by mutant SOD1 is responsible for the cellular toxicity, but how mutant SOD1 kills motoneurons is unclear. In transgenic mouse models of ALS, mitochondrial degeneration occurs early, before disease onset, raising the question of how mutant SOD1 damages mitochondria. Here we investigate the intracellular localization of SOD1 in the CNS to determine whether SOD1 is present in mitochondria, where it could directly damage this organelle. We show that endogenous mouse SOD1, wild-type human, and mutant human SOD1 (G93A), when expressed as transgenes, are colocalized with mitochondria in spinal cord by immunofluorescence confocal microscopy. By immunoelectron microscopy, we show that SOD1 is present within mitochondria at similar concentrations as in the cytoplasm. Thus SOD1, in addition to being a cytosolic enzyme, is present inside mitochondria in the CNS. PMID- 11886901 TI - Arthroscopic synovectomy of the elbow in rheumatoid arthritis. AB - BACKGROUND: The purpose of this study was to investigate the results of arthroscopic synovectomy for the treatment of elbows affected by rheumatoid arthritis. METHODS: Arthroscopic synovectomy was performed on twenty-nine elbows (twenty-seven patients) between 1984 and 1996. Twenty-one elbows (twenty patients), followed for a minimum of forty-two months, were evaluated clinically with use of the Mayo elbow performance score and radiographic findings. The mean duration of follow-up was ninety-seven months. With use of the system of Larsen et al., we classified all elbows into three groups--Grades 1 and 2, Grade 3, and Grade 4--according to the preoperative radiographic findings. These groups were then compared. RESULTS: The mean Mayo elbow performance score improved from 48.3 points preoperatively to 77.5 points (an excellent result in two elbows, a good result in thirteen, a fair result in six, and a poor result in none) at two years after the operation and 69.8 points (an excellent result in two elbows, a good result in seven, a fair result in seven, and a poor result in five) at the final follow-up evaluation. The mean score for pain improved from 9.3 points preoperatively to 31.4 points at two years after the operation and 27.9 points at the final follow-up evaluation. Clinically apparent synovitis recurred in five of the twenty-one elbows, and two of the five required total elbow arthroplasty. Among the three groups, only elbows with Larsen Grade-1 or 2 arthritis had a favorable long-term result with regard to total function. The postoperative results were unsatisfactory for Larsen Grade-4 elbows. CONCLUSIONS: Arthroscopic synovectomy in an elbow affected by rheumatoid arthritis is a reliable procedure that can alleviate pain. Our results suggest that one of the most favorable indications for arthroscopic synovectomy is a preoperative radiographic rating of Grade 1 or 2. PMID- 11886902 TI - Impact of cost reduction programs on short-term patient outcome and hospital cost of total knee arthroplasty. AB - BACKGROUND: During the 1990s, cost reduction programs were developed to decrease the hospital cost of total knee arthroplasty. The purpose of this study was to evaluate the impact of hospital cost reduction programs for total knee arthroplasty on patient outcome at our hospital. METHODS: We evaluated 159 patients who had undergone unilateral primary total knee arthroplasty for the treatment of osteoarthritis at the Lahey Clinic. The results of fifty-six knee replacements performed in 1992 without a clinical pathway or a knee-implant standardization program (the control group) were compared with the results of 103 knee replacements performed in 1995 with a clinical pathway and a knee-implant standardization program (the study group). Before the operation, the two patient populations were similar in terms of age, pain score on a visual analog scale, and clinical knee scores; the groups were also similar with regard to the surgical approach and the time in the operating room. The minimum duration of follow-up was eight years for the control group and five years for the study group. RESULTS: All patients in both groups had excellent relief of pain and improvement in function. There were no differences in clinical outcome between the patient groups. The rate of patient satisfaction was 98% in the control group and 99% in the study group. Implementation of the clinical pathway was associated with a reduction in the average length of the stay in the hospital from 6.79 days in 1992 to 4.16 days in 1995. Implementation of the knee-implant standardization program was associated with increased use of all-polyethylene tibial components in 1995. Hospital cost adjusted for medical inflation was reduced 19% with the implementation of the clinical pathway and the knee-implant standardization program. CONCLUSIONS: The clinical pathway and the knee-implant standardization program reduced resource utilization and hospital cost for total knee arthroplasty without affecting short-term patient outcome in our hospital. Orthopaedic surgeons should carefully evaluate cost reduction programs, which may affect their patients, in order to maintain high-quality orthopaedic care and consistently successful patient outcomes. PMID- 11886903 TI - Closed reduction of colles fractures: comparison of manual manipulation and finger-trap traction: a prospective, randomized study. AB - BACKGROUND: An optimal outcome of closed treatment of a Colles fracture may depend on accurate reduction and adequate immobilization. It has been suggested that the use of finger-trap traction results in a better reduction and a lower rate of redisplacement than manual manipulation does, but to our knowledge these concepts have never been evaluated scientifically. We compared these two methods in a prospective, randomized controlled trial. METHODS: Two hundred and twenty three patients with 225 displaced Colles-type fractures were randomized to treatment with closed reduction with either finger-trap traction (112 patients) or manual manipulation (111 patients). The fractures were assessed radiographically by measurement of the radial angle, dorsal tilt, and radial shortening before reduction, immediately after reduction, and at one and five weeks after reduction. RESULTS: The groups were comparable with regard to age, sex, side of injury, fracture grade, and amount of displacement at presentation. No significant differences were found between the alignment of the fractures in the two treatment groups at any time. With dorsal tilt of <10 degrees and radial shortening of <5 mm considered acceptable, the two techniques both produced an 87% rate of satisfactory reductions. However, the percentages of fractures in an acceptable alignment were only 57% and 50% at one week after finger-trap traction and manual manipulation, respectively, and only 27% and 32% at five weeks. The failure rates did not differ significantly between the two groups. CONCLUSIONS: The two methods of fracture reduction did not differ with regard to the eventual position of the fracture or the rate of failure. Although closed reduction was successful for the majority of fractures, most redisplaced substantially during the period of cast immobilization. PMID- 11886904 TI - Analysis of vertebral morphology in idiopathic scoliosis with use of magnetic resonance imaging and multiplanar reconstruction. AB - BACKGROUND: Several studies have provided data on the vertebral morphology of normal spines, but there is a paucity of data on the vertebral morphology in patients with idiopathic scoliosis. METHODS: The morphology of the pedicles and bodies of 307 vertebrae as well as the distance between the pedicles and the dural sac (the epidural space) in twenty-six patients with right-sided thoracic idiopathic scoliosis were analyzed with use of magnetic resonance imaging and multiplanar reconstruction. RESULTS: A distinct vertebral asymmetry was found at the apical region of the thoracic curves, with significantly thinner pedicles on the concave side than on the convex side (p < 0.05). The degree of intravertebral deformity diminished farther away from the apex, with vertebral symmetry restored at the neutral level. In the thoracic spine, the transverse endosteal width of the apical pedicles measured between 2.3 mm and 3.2 mm on the concave side and between 3.9 mm and 4.4 mm on the convex side (p < 0.05). In the lumbar spine, the pedicle width measured between 4.6 mm at the cephalad part of the curve and 7.9 mm at the caudad part of the curve. The chord length and the pedicle length gradually increased from 34 mm and 18 mm, respectively, at the fourth thoracic vertebra to 51 mm and 25 mm, respectively, at the third lumbar vertebra. The transverse pedicle angle measured 15 in the cephalad aspect of the thoracic spine, decreased to 7 at the twelfth thoracic vertebra, and increased again to 16 at the fourth lumbar vertebra. The width of the epidural space was <1 mm at the thoracic apical vertebral levels and averaged 1 mm at the lumbar apical vertebral levels on the concave side, whereas it was between 3 mm and 5 mm on the convex side (p < 0.05). CONCLUSION: Idiopathic scoliosis is associated with distinctive intravertebral deformity, with smaller pedicles on the concave side and a shift of the dural sac toward the concavity. PMID- 11886905 TI - Analysis of polyethylene thickness of tibial components in total knee replacement. AB - BACKGROUND: Excessive wear of the polyethylene bearing surfaces of tibial components has become an important factor in early failure of total knee arthroplasty. Inadequate thickness of the polyethylene insert is one cause of excessive wear, and various minimum thicknesses have been recommended in order to reduce contact stresses within the polyethylene. However, the true thicknesses of modular polyethylene tibial inserts typically are not stated accurately by the manufacturers in their product information. The purpose of this study was to determine whether the information about the thickness of tibial inserts supplied by the manufacturers is adequate. METHODS: Five of the thinnest available polyethylene tibial inserts from five different manufacturers were selected. The minimum thickness of each was measured with use of a Sigma electronic micrometer comparator to an accuracy of +/-0.005 mm. RESULTS: The stated thicknesses of the polyethylene tibial inserts were 8, 9, and 10 mm, values that differed markedly from the actual minimum thicknesses, which ranged from 5.5722 to 6.2048 mm (mean values). CONCLUSION: The thickness of polyethylene tibial inserts has been implicated as a potential cause of excessive wear and early failure of total knee replacements. This paper highlights the fact that the information supplied by the manufacturers is inaccurate and potentially misleading; in one case, the true thickness was much less than the recommended minimum thickness. We recommend that the minimum thickness of the tibial components as well as the combined thickness of the polyethylene insert and the metal tibial tray be specified in the product information and on the packaged insert. PMID- 11886906 TI - Treatment of reverse oblique and transverse intertrochanteric fractures with use of an intramedullary nail or a 95 degrees screw-plate: a prospective, randomized study. AB - BACKGROUND: Intertrochanteric fractures are composed of different anatomic patterns that vary in their degree of stability following open reduction and internal fixation. A particularly unstable group is classified as AO/OTA 31-A3, with the fracture pattern described as reverse oblique or transverse. The purpose of this study was to compare the results of intramedullary fixation with those of plate fixation for these intertrochanteric fractures in elderly patients. METHODS: Thirty-nine elderly patients with AO/OTA 31-A3 intertrochanteric fractures of the femur were randomized into two treatment groups and were followed for a minimum of one year. The nineteen patients in Group I were treated with a 95 fixed-angle screw-plate (Dynamic Condylar Screw), and the twenty patients in Group II were treated with an intramedullary nail (Proximal Femoral Nail). The treatment groups were comparable with regard to all demographic and injury variables. RESULTS: Patients treated with an intramedullary nail had shorter operative times, fewer blood transfusions, and shorter hospital stays compared with those treated with a 95 screw-plate. Implant failure and/or nonunion was noted in seven of the nineteen patients who had been treated with the 95 screw-plate. Only one of the twenty fractures that had been treated with an intramedullary nail did not heal. CONCLUSION: The results of our study support the use of an intramedullary nail rather than a 95 screw-plate for the fixation of reverse oblique and transverse intertrochanteric fractures in elderly patients. PMID- 11886907 TI - Operative treatment of elbow contracture in patients twenty-one years of age or younger. AB - BACKGROUND: Elbow contracture is a recognized sequela of traumatic and developmental elbow disorders, but little information is available regarding the surgical treatment of elbow stiffness in the pediatric population. METHODS: Thirty-seven patients who had had open surgical release of an elbow contracture at a mean age of sixteen years (range, ten to twenty years) were retrospectively studied after a mean duration of follow-up of fifteen months (range, six to forty four months). The elbow contracture was posttraumatic in twenty-eight patients. The operation consisted of a capsular release with removal of osseous impediments to motion as necessary. No patient had muscle or tendon-lengthening. RESULTS: The total arc of motion improved from a mean of 66 preoperatively to a mean of 94 postoperatively; however, only twenty-eight patients (76%) had an improvement of 10 and only seventeen (46%) achieved a functional arc of motion of 100 (from 30 to 130 ). Two patients lost motion after surgery. These results are less favorable than the results of previous studies of both pediatric and adult patients. Patients in whom the contracture had been caused by a simple dislocation of the elbow or an extra-articular fracture tended to have better results than those in whom the contracture was due to other causes. CONCLUSIONS: The results of surgical treatment of elbow stiffness in pediatric patients are less favorable and less predictable than those in adult patients. PMID- 11886908 TI - The quality of reporting of randomized trials in the Journal of Bone and Joint Surgery from 1988 through 2000. AB - BACKGROUND: The purpose of this study was threefold: (1) to determine the scientific quality of published randomized trials in the American Volume of The Journal of Bone and Joint Surgery from 1988 through 2000, (2) to identify predictors of study quality, and (3) to evaluate inter-rater agreement in the scoring of study quality with use of a simple scale. METHODS: Hand searches of The Journal of Bone and Joint Surgery were conducted in duplicate to identify randomized clinical trials. Of 2468 studies identified, seventy-two (2.9%) met all eligibility criteria. Two investigators each assessed the quality of the study under blinded conditions and abstracted relevant data. RESULTS: The mean score (and standard error) for the quality of the seventy-two randomized trials was 68.1% plus minus 1.6%; 60% (forty-three) scored <75%. Drug trials had a significantly higher mean quality score than did surgical trials (72.8% compared with 63.9%, p < 0.05). Regression analysis revealed that cited affiliation with an epidemiology department and cited funding were associated with higher quality scores. Failure to conceal randomization, to blind outcome assessors, and to describe why patients were excluded resulted in significantly lower quality scores (p < 0.05), more than the 5% decrease expected by removal of each item. A priori calculations of sample size were rarely performed in the reviewed studies, and only 2% of the studies with negative results included a post hoc power analysis. The Detsky quality scale met accepted standards of interobserver reliability (kappa, 0.87; 95% confidence interval, 0.70 to 0.95). CONCLUSIONS: Few studies published in The Journal of Bone and Joint Surgery were randomized trials. More than half of the trials were limited by a lack of concealed randomization, lack of blinding of outcome assessors, or failure to report reasons for excluding patients. Application of standardized guidelines for the reporting of clinical trials in orthopaedics should improve quality. PMID- 11886909 TI - Evaluation of the forearm in untreated adult subjects with multiple hereditary osteochondromatosis. AB - BACKGROUND: Limb-length discrepancy or angular deformities as a result of altered bone growth may lead to a decreased range of motion and impaired function as well as premature osteoarthritis in patients with multiple hereditary osteochondromatosis. The purpose of this study was to describe the function of the forearm in untreated patients in order to facilitate comparison with studies of the results of treatment of this condition. METHODS: The medical records of fifty-one pediatric patients were identified and served as the basis for identifying a cohort of adult relatives with the disorder. Participants were asked about pain and limitations in vocational or recreational activities and about concerns with regard to cosmetic appearance. Radiographs of the forearm and wrist were made to quantify the deformity. Functional outcome was assessed on the basis of a comparison with the norms for grip and pinch strength and for scores on the hand function test of Jebsen et al. Limitations in the range of motion of the upper extremities were converted to standard impairment ratings. RESULTS: Participants included twenty-two men and seventeen women with an average age of forty-two years (range, twenty to eighty years). Most of the patients were employed in careers of their choice, with only five (13%) indicating that they were limited in any way in the performance of their jobs. Twenty-six subjects (67%) participated in recreational activities, and sixty-eight arms (88%) were reported to be free of pain. Objective measurement of function demonstrated greater disability than that found from subjective reporting. Fourteen arms had an impairment rating of >10%, while twenty had decreased pinch strength and sixteen had decreased grip strength. Ten arms (13%) had decreased hand function according to the hand test of Jebsen et al. Radiographic evaluation demonstrated osteoarthritic changes in three limbs. CONCLUSIONS: Affected individuals had definite decreases in hand and wrist function, yet these did not result in major increases in pain or in limitations in daily work and recreation. This cohort provides a basis for comparison with the results of operative treatment in affected individuals with multiple hereditary osteochondromatosis. PMID- 11886910 TI - Eccentric rotational acetabular osteotomy for acetabular dysplasia: follow-up of one hundred and thirty-two hips for five to ten years. AB - BACKGROUND: Eccentric rotational acetabular osteotomy for the operative treatment of acetabular dysplasia consists of a spherical but eccentric osteotomy and rotation of the acetabulum that moves the center of rotation of the head of the femur medially and distally. No bone graft is needed. The reorientation of the acetabular fragment not only improves acetabular coverage but also restores the center of rotation of the subluxated hip. The purpose of this paper was to describe eccentric rotational acetabular osteotomy for the treatment of acetabular dysplasia and to evaluate its clinical and radiographic outcomes. METHODS: We performed this procedure consecutively in 132 hips in 126 patients with dysplasia of the hip. Eighteen hips had no osteoarthritis, fifty-three had early osteoarthritis, and sixty-one had advanced osteoarthritis. Seven patients were male, and 119 were female. The average age was 36.5 years at the time of the index operation, and the average duration of follow-up was 7.5 years. Twenty three hips in twenty-two patients were also treated with intertrochanteric valgus osteotomy to further improve joint congruency at the time of the acetabular osteotomy. RESULTS: The average preoperative Harris hip score of 71 points improved to an average score of 89 points at the time of the latest follow-up. The average center-edge angle improved from 0 to 36 . An apparent change in the stage of the arthritis was observed in seven hips (5%), one of which had had early-stage disease and six of which had had advanced disease preoperatively. CONCLUSIONS: Eccentric rotational acetabular osteotomy appears to be a good treatment option for young patients with either early or advanced hip osteoarthritis secondary to dysplasia. PMID- 11886912 TI - Surgical correction of the snapping iliopsoas tendon in adolescents. AB - BACKGROUND: There have been very few reports regarding symptomatic snapping of the iliopsoas tendon, and none of those reports have dealt exclusively with an adolescent population. We report our experience with the surgical treatment of this entity in a group of patients who had an average age of fifteen years. METHODS: Nine adolescent patients (eleven hips) underwent fractional lengthening of the iliopsoas tendon at the musculotendinous junction because of persistent painful snapping of the hip. A modified iliofemoral approach to the iliopsoas tendon was used. The diagnosis in all cases was made on the basis of the history and a physical examination. Plain radiographs were made for all patients to rule out an osseous intra-articular loose body. Follow-up consisted of personal interviews and physical examinations performed at least two years postoperatively. RESULTS: Preoperatively, all patients had audible snapping with pain localized to the anterior part of the groin. The average duration of symptoms was 2.3 years. Prior to the onset of symptoms, all but one of the patients had been involved in competitive athletic activities. Postoperatively, all patients were able to return to the preoperative level of activity without subjective weakness. The average duration of postoperative follow-up was four years. Hip-flexion strength was noted to be nearly equal to that on the contralateral side. All patients reported that they would have the operation again under similar circumstances. One patient had recurrent snapping but stated that it was less frequent and less painful than the preoperative snapping. Two patients had transient sensory loss in the anterolateral aspect of the thigh. CONCLUSIONS: We conclude that fractional lengthening of the iliopsoas tendon at the musculotendinous junction is an effective and safe approach for adolescent patients with persistent symptomatic snapping of the iliopsoas tendon that is unresponsive to conservative measures. PMID- 11886911 TI - Nitric oxide and prostaglandin E2 production in response to ultra-high molecular weight polyethylene particles depends on osteoblast maturation state. AB - BACKGROUND: Recent studies have shown that osteoblast-like cells respond directly to ultra-high molecular weight polyethylene particles in culture, suggesting that they may be involved in aseptic loosening of endoprostheses. We tested the hypothesis that the state of cell maturation plays a role in the response of osteogenic cells to ultra-high molecular weight polyethylene particles. METHODS: MG63 cells (immature osteoblast-like cells), OCT-1 cells (mature secretory osteoblast-like cells), and MLO-Y4 cells (osteocyte-like cells) were treated for twenty-four hours with commercial ultra-high molecular weight polyethylene particles with an average diameter of 1 mm. The effect of particle treatment on cell proliferation was assessed by measuring the number of cells, whereas the effects on differentiation and local factor production were assessed by measuring the production of osteocalcin, prostaglandin E2, and nitric oxide. The effect of particles on apoptosis was also evaluated. RESULTS: The addition of ultra-high molecular weight polyethylene particles increased the number of MG63 cells, did not affect the number of OCT-1 cells, and led to a decrease in the number of MLO Y4 cells. The observed changes in cell number were not due to programmed cell death, as no more than 3% of the cells in cultures treated with the highest concentration of particles were undergoing apoptosis. Osteocalcin production was not affected by the addition of particles. Prostaglandin E2 production was increased in all three types of cultures, but the effect was greatest in OCT-1 cell cultures, as was the absolute amount of prostaglandin E2 produced. Nitric oxide production was unaffected in MG63 cell cultures, but it was stimulated in OCT-1 and MLO-Y4 cell cultures. CONCLUSIONS: The results of the present study support the hypothesis that osteoblast cell maturation state plays an important role in the response to ultra-high molecular weight polyethylene particles and that the terminally differentiated osteocyte may be involved in the bone response to wear debris in vivo. PMID- 11886913 TI - Cementless total hip arthroplasty with a tapered, rectangular titanium stem and a threaded cup: a minimum ten-year follow-up. AB - BACKGROUND: We report the results of cementless total hip arthroplasty with a tapered, rectangular titanium stem that was introduced in 1979 and continues to be used today with only minor changes. The aim of the design is to achieve primary stability to resist rotational and axial forces through precision rasping and press-fit implantation of a tapered, rectangular femoral component. METHODS: Between October 1986 and November 1987, 208 total hip arthroplasties with insertion of a tapered, rectangular titanium stem and a threaded cup without cement were performed in 200 consecutive patients (average age, sixty-one years; range, twenty-two to eighty-four years). RESULTS: At the time of the latest follow-up, fifty-one patients (fifty-two hips) had died and sixteen patients had been lost to follow-up, leaving 133 patients. Twelve hips had been revised, two in patients who subsequently died, leaving 123 living patients without revision. The median follow-up time was 120.7 months. Five cups needed revision surgery because of aseptic loosening; two, because of massive polyethylene wear; one, because of posttraumatic migration; and one, because of breakage. Three femoral stems were revised: one because of malpositioning (the reoperation was done five days after implantation); one, because of infection; and the third, after multiple failed acetabular revisions. The mean Harris hip score for the patients who did not have revision was 85.4 points (range, 46 to 100 points) at the time of the latest follow-up. Four patients (3%) complained of thigh pain that was not associated with another disorder. According to the criteria of Engh et al., all femoral implants were graded as stable bone-ingrown. The probability of survival of both the femoral and the acetabular component at ten years, with any revision as the end point, was 0.92 (95% confidence interval, 0.88 to 0.97). The probability of survival of the cup was 0.93 (95% confidence interval, 0.89 to 0.97), and that of the stem was 0.99 (95% confidence interval, 0.97 to 1.00). CONCLUSIONS: The results of arthroplasty with a tapered, rectangular titanium stem combined with a conical threaded cup inserted without cement were excellent at a minimum of ten years. Our data suggest that femoral stem fixation continues to be secure, while the threaded cup is prone to aseptic loosening. PMID- 11886914 TI - Fibrolipomatous hamartoma in the foot: magnetic resonance imaging and surgical treatment: a report of two cases. PMID- 11886915 TI - Thoracic outlet syndrome treated by double osteotomy of a clavicular malunion: a case report. PMID- 11886916 TI - Pseudomonas aeruginosa osteomyelitis of both ischia: a case report. PMID- 11886917 TI - Ewing sarcoma in an octogenarian: a case report. PMID- 11886918 TI - Fungal infection following replacement of the anterior cruciate ligament: a case report. PMID- 11886919 TI - Bone-grafting and bone-graft substitutes. PMID- 11886921 TI - Alfred R. Shands, Jr., lecture: our humanitarian orthopaedic opportunity. PMID- 11886922 TI - Application of the Consolidated Standards of Reporting Trials (CONSORT) in the Fracture Care Literature. PMID- 11886923 TI - Two-stage reimplantation for the salvage of infected total knee arthroplasty. 1983. PMID- 11886924 TI - Diagnosing atlantoaxial impaction and basilar invagination. PMID- 11886925 TI - Differences in orthopaedic training in Canada and the United kingdom. PMID- 11886926 TI - Orthopaedic training and practice in the Canadian Health-Care System. PMID- 11886927 TI - Fixation of reverse obliquity fractures of the subtrochanteric and intertrochanteric regions of the femur. PMID- 11886928 TI - The biomechanics of glenohumeral stability. PMID- 11886929 TI - Magnetic resonance imaging or ultrasound in assessment of the interosseous membrane of the forearm. PMID- 11886930 TI - Complications of total knee arthroplasty after open reduction and internal fixation of fractures of the tibial plateau. PMID- 11886931 TI - Trauma and orthopaedic training in the United Kingdom. PMID- 11886932 TI - What's new in foot and ankle surgery. PMID- 11886934 TI - Hepatic insulin resistance in obese non-diabetic subjects and in type 2 diabetic patients. AB - OBJECTIVE: Obese non-diabetic patients are characterized by an extra-hepatic insulin resistance. Whether obese patients also have decreased hepatic insulin sensitivity remains controversial. RESEARCH METHODS AND PROCEDURES: To estimate their hepatic insulin sensitivity, we measured the rate of exogenous insulin infusion required to maintain mildly elevated glycemia in obese patients with type 2 diabetes, obese non-diabetic patients, and lean control subjects during constant infusions of somatostatin and physiological low-glucagon replacement infusions. To account for differences in insulin concentrations among the three groups of subjects, an additional protocol was also performed in healthy lean subjects with higher insulin infusion rates and exogenous dextrose infusion. RESULTS: The insulin infusion rate required to maintain glycemia at 8.5 mM was increased 4-fold in obese patients with type 2 diabetes and 1.5-fold in obese non diabetic patients. The net endogenous glucose production (measured with 6,6 (2)H(2)-glucose) and total glucose output (measured with 2-(2)H(1)-glucose) were approximately 30% lower in the patients than in the lean subjects. Net endogenous glucose production and total glucose output were both markedly increased in both groups of obese patients compared with lean control subjects during hyperinsulinemia. DISCUSSION: Our data indicate that both obese non-diabetic and obese type 2 diabetic patients have a blunted suppressive action of insulin on glucose production, indicating hepatic and renal insulin resistance. PMID- 11886935 TI - Probability of adult overweight and risk change during the BMI rebound period. AB - OBJECTIVE: To develop a probability chart of adult overweight based on childhood body mass index (BMI) values and to evaluate the BMI change during the BMI rebound period during childhood, in different populations, with the use of risk function curves. RESEARCH METHODS AND PROCEDURES: A longitudinal growth study of 3650 full-term healthy Swedish children followed from birth to 18 years of age. Weight and height values of our subjects were obtained. RESULTS: A probability chart for reaching a BMI > 23 kg/m(2) at 18 years of age was constructed for boys and girls. For example, a BMI of 18 kg/m(2) at 4 years of age is associated with 0.70 probability of attaining a BMI > 23 kg/m(2) at 18 years of age in boys; a BMI of 16 kg/m(2) at 4 years of age leads to 0.40 probability of having a BMI > 23 kg/m(2) at 18 years of age in girls. Children with an obvious BMI rebound before 8 years of age have a high risk of being overweight at 18 years of age. There is a clear trend of BMI increase from the 1970s to the 1990s in U.S. children from a parallel dataset, and Hispanic children are at the highest risk of adult overweight. DISCUSSION: The probability chart for adult overweight developed here provides a functional method of defining childhood obesity that is based on the risk of long-term ill health rather than on a certain statistical cut-off point. It will help pediatricians or healthcare workers identify those children who are at a high risk of becoming overweight in adulthood, which will allow clinical intervention at younger ages. PMID- 11886936 TI - Ethnic differences in physical activity and inactivity patterns and overweight status. AB - OBJECTIVE: To examine the relationship between physical activity and inactivity patterns and overweight in U.S. adolescents using baseline and 1-year change in activity and inactivity data. RESEARCH METHODS AND PROCEDURES: Nationally representative data from 12,759 participants (6997 non-Hispanic whites, 2676 non Hispanic blacks, 2185 Hispanics, and 901 Asians) in the National Longitudinal Study of Adolescent Health (1995 and 1996). Data on moderate to vigorous and low intensity physical activity, TV/video viewing, and video game/computer use were obtained from questionnaires. Multivariate models assessed the association of overweight (body mass index > or = 95th percentile Centers for Disease Control and Prevention/National Center for Health Statistics 2000 curves) with initial (and 1-year change) activity and inactivity levels, controlling for age, ethnicity, socioeconomic status, urban residence, cigarette smoking, and region of residence. RESULTS: Overweight prevalence was positively associated with high level TV/video viewing among white boys (odds ratio [OR] = 1.52; 95% confidence interval [1.08 to 2.14]) and girls (OR = 2.45 [1.51 to 3.97]). The odds of overweight decreased with high levels of moderate to vigorous physical activity among white boys (OR = 0.81 [0.76 to 0.87]), non-Hispanic black boys (OR = 0.86 [0.76 to 0.98]) and girls (OR = 0.88 [0.78 to 0.99]), and Hispanic boys (OR = 0.90 [0.83 to 0.97]) and girls (OR = 0.91 [0.84 to 0.99]). DISCUSSION: Predicted probabilities generated from the logistic regression models, which examined the experimental effects of altering hours of TV/video viewing and bouts of moderate to vigorous physical activity, show lower overweight among adolescents who watched less TV per week combined with frequent moderate to vigorous physical activity than those who watched more TV per week combined with fewer bouts of weekly moderate to vigorous physical activity. Predicted probabilities suggest important sex and ethnic differences in these associations. PMID- 11886937 TI - Validation and calibration of physical activity monitors in children. AB - OBJECTIVE: This study was designed to validate accelerometer-based activity monitors against energy expenditure (EE) in children; to compare monitor placement sites; to field-test the monitors; and to establish sedentary, light, moderate, and vigorous threshold counts. RESEARCH METHODS AND PROCEDURES: Computer Science and Applications Actigraph (CSA) and Mini-Mitter Actiwatch (MM) monitors, on the hip or lower leg, were validated and calibrated against 6-hour EE measurements by room respiration calorimetry, activity by microwave detector, and heart rate by telemetry in 26 children, 6 to 16 years old. During the 6 hours, the children performed structured activities, including resting metabolic rate (RMR), Nintendo, arts and crafts, aerobic warm-up, Tae Bo, treadmill walking and running, and games. Activity energy expenditure (AEE) computed as EE - RMR was regressed against counts to derive threshold counts. RESULTS: The mean correlations between EE or AEE and counts were slightly higher for MM-hip (r = 0.78 +/- 0.06) and MM-leg (r = 0.80 +/- 0.05) than CSA-hip (r = 0.66 +/- 0.08) and CSA-leg (r = 0.73 +/- 0.07). CSA and MM performed similarly on the hip (inter instrument r = 0.88) and on the lower leg (inter-instrument r = 0.89). Threshold counts for the CSA-hip were <800, <3200, <8200, and > or = 8200 for sedentary, light, moderate, and vigorous categories, respectively. For the MM-hip, the threshold counts were <100, <900, <2200, and > or = 2200, respectively. DISCUSSION: The validation of the CSA and MM monitors against AEE and their calibration for sedentary, light, moderate, and vigorous thresholds certify these monitors as valid, useful devices for the assessment of physical activity in children. PMID- 11886938 TI - Does ethnicity influence body-size preference? A comparison of body image and body size. AB - OBJECTIVE: Obesity is most common in the United States among women of ethnic minority groups (black and Hispanic). Researchers have hypothesized that these subcultures are more accepting of overweight figures. The purpose of this study was to examine body image and body size assessments in a large community sample of men and women. RESEARCH METHODS AND PROCEDURES: Participants were 801 women and 428 men: 23% Asian, 45% Hispanic, 17% black, and 15% white. The figure rating scale was used to rate: body dissatisfaction, attractive male and female shapes, acceptable female size, and perceptions of underweight to obese female figures. RESULTS: Controlling for age, education, and body weight, no ethnic differences were found for men. Asian women reported less body dissatisfaction than the other groups. Women were more dissatisfied with their size than men and chose thinner female figures as attractive and acceptable. DISCUSSION: Ethnicity, independent of age, education, and body weight, does not influence preference for female and male shapes or tolerance for obesity. PMID- 11886940 TI - Regulation of body weight and carcass composition by sibutramine in rats. AB - OBJECTIVE: We examined the effectiveness of sibutramine to modulate food intake and body composition in rats with two levels of adiposity imposed by the duration of their maintenance on a moderate-fat diet. RESEARCH METHODS AND PROCEDURES: Male Sprague--Dawley rats were fed a 32% fat diet from weaning until 2 or 4 months of age, at which point, body fat was either 15% or 25%, respectively, as measured by DXA. Sibutramine (0.6 or 2 mg/kg, orally) was then given daily for 2 weeks. RESULTS: Food intake and body weight decreased acutely in a dose-related manner in both groups with sibutramine treatment. In all rats, food intake suppression was attenuated after multiple days of sibutramine. Both 15%- and 25% fat rats had a persistent decrease in weight gain over the 2-week period in response to sibutramine. The older, 25%-fat rats were more sensitive to sibutramine than the younger, 15%-fat rats with regard to the magnitude of overall food intake inhibition, decrease in body weight gain, and caloric efficiency. Despite these differences, sibutramine produced the same relative reductions in fat mass and had no effect on lean mass in the two groups. DISCUSSION: Thus, sibutramine produced equivalent efficacy on carcass fat loss in both groups, despite less inhibition of feeding and body weight gain in leaner rats. Whether these changes are a result of the leaner rats being younger and on a steeper growth curve compared with older, fatter rats or whether this is a direct function of their level of adiposity remains to be determined. PMID- 11886939 TI - Estradiol may limit lipid oxidation via Cpt 1 expression and hormonal mechanisms. AB - OBJECTIVE: Evidence indicates that estrogen depresses hepatic lipid oxidation. We tested the hypothesis that estradiol (E(2)) treatment depresses transcription of carnitine palmitoyltransferase-1 (Cpt 1) mRNA and increases adiposity. RESEARCH METHODS AND PROCEDURES: Six ovariectomized female rats were given a subcutaneous pellet of E(2) (5 mg/d), and six were given placebo. Rats were pair-fed by group for 18 days. Body composition was assessed chemically: mRNA for liver Cpt 1, adipose tissue uncoupling protein-2 (Ucp 2), and quadriceps Ucp 3 by Northern analysis; serum glucose, triglycerides (TGs), and free fatty acids by standard techniques; and serum insulin and glucagon by radioimmunoassay. RESULTS: E(2) treated rats lost more weight than placebo-treated rats (37.3 +/- 6.0 vs. 16.2 +/ 2.6 g, p < 0.01), but did not differ in final carcass composition (adjusted for eviscerated body mass). E(2)-treated rats had lower liver Cpt 1 (p < 0.001) and skeletal muscle Ucp 3 (p < 0.05) mRNA and lower concentrations of glucose, glucagon, and free fatty acids (p < 0.05). E(2)-treated rats tended to have higher insulin (p = -0.067) and TG (p = 0.097). TG tended to be correlated with Cpt 1 mRNA (r = -0.56 and p = 0.07). DISCUSSION: These results suggest that, although E(2) is likely to suppress lipid oxidation and promote TG synthesis, these effects are not manifested in a relative increase in carcass adiposity after 18 days of treatment, at least under conditions of negative energy balance. The possible role of E(2)-mediated changes in insulin and glucagon secretion on hepatic substrate metabolism warrants further study. PMID- 11886941 TI - Chronic application of MTII in a rat model of obesity results in sustained weight loss. AB - OBJECTIVE: To examine the effects of a cafeteria diet and a chronic treatment with melanocortin agonist (MTII) on mature weight-stable female rats. RESEARCH METHODS AND PROCEDURES: Ex-breeder Chbb:Thom rats (350 to 400 g) were divided into two groups: highly palatable food (HPF) and normal rat chow (RC). Both groups had ab libitum access to rat chow. The HPF group had access to chocolate bars, cookies, cheese, and nuts (approximately 20 g/d). After 21 days, the rats in each group were then divided into control and treated groups. Mini-pumps delivering saline or MTII (1 mg/kg per day) for minimally 28 days were implanted. Oxygen consumption was measured for 17 days in a second group of rats implanted with mini-pumps containing MTII (1 mg/kg per day) or saline. RESULTS: HPF rats ate less (<50%) rat chow than RC rats. After 20 days, the HPF group had reached a plateau and weighed significantly more (p < 0.005) than the RC group (411.7 +/- 9.3 g; n = 17 vs. 365.1 +/- 9.4 g; n = 16). HPF rats and RC rats receiving MTII reduced their pellet intake and body weight in the initial 2 weeks of treatment (day 14, RC-saline: -1.6 +/- 1.8 g; RC-MTII, -22.5 +/- 3.7 g; HPF-saline, -7.1 +/ 1.7 g; HPF-MTII, -30.7 +/- 4.8 g). Subsequently, pellet intake returned to pre implantation values, although body weights remained reduced in both HPF and RC groups. Oxygen consumption was increased in rats treated with MTII. DISCUSSION: This suggests that MTII initially reduced body weight by limiting food intake; however, maintenance of weight is most likely due to increased energy expenditure under conditions of normal and highly palatable diets in mature animals. PMID- 11886943 TI - The human obesity gene map: the 2001 update. AB - This report constitutes the eighth update of the human obesity gene map, incorporating published results up to the end of October 2001. Evidence from the rodent and human obesity cases caused by single-gene mutations, Mendelian disorders exhibiting obesity as a clinical feature, quantitative trait loci (QTLs) uncovered in human genome-wide scans and in crossbreeding experiments in various animal models, association and linkage studies with candidate genes and other markers is reviewed. The human cases of obesity related in some way to single-gene mutations in six different genes are incorporated. Twenty-five Mendelian disorders exhibiting obesity as one of their clinical manifestations have now been mapped. The number of different QTLs reported from animal models currently reaches 165. Attempts to relate DNA sequence variation in specific genes to obesity phenotypes continue to grow, with 174 studies reporting positive associations with 58 candidate genes. Finally, 59 loci have been linked to obesity indicators in genomic scans and other linkage study designs. The obesity gene map depicted in Figure 1 reveals that putative loci affecting obesity related phenotypes can be found on all chromosomes except chromosome Y. A total of 54 new loci have been added to the map in the past 12 months, and the number of genes, markers, and chromosomal regions that have been associated or linked with human obesity phenotypes is now above 250. Likewise, the number of negative studies, which are only partially reviewed here, is also on the rise. PMID- 11886942 TI - Metabolic characteristics and body composition in a model of anti-obese rats (Lou/C). AB - OBJECTIVE: The aims of this study were to investigate some features of the metabolic profile and the body composition of male Lou/C rats and to examine whether these characteristics are strictly related to the food-intake reduction. RESEARCH METHODS AND PROCEDURES: Fourteen-week-old male Lou/C rats were compared with age-matched male Wistar rats fed ad libitum (WAL) and another group of male Wistar rats whose food was chronically restricted (WFR) to the same amount as the Lou/C rats from weeks 3 to 14. RESULTS: Food intake and body weight were significantly (p < 0.01) reduced in Lou/C compared with WAL rats, whereas these reductions were perfectly reproduced in WFR rats. Lou/C rats demonstrated lower relative weights of retroperitoneal (0.97 +/- 0.07 vs. 1.67 +/- 0.16 and 1.88 +/- 0.15 g/100 g body) and epididymal (1.01 +/- 0.02 vs. 1.62 +/- 0.12 and 1.80 +/- 0.11 g/100g body) fat depots than did the two other groups and no decrease in the percentage of carcass proteins, which was observed in the WFR rats. In addition, compared with the WFR group, the Lou/C rats showed lower plasma glucose levels (3.65 +/- 0.14 vs. 4.72 +/- 0.15 and 4.7 +/- 0.19 mM); a tendency (p < 0.1) for lower liver glycogen concentrations; and similar levels of glycerol, free fatty acids, and beta-hydroxybutyrate concentrations. Epinephrine and the relative weight of the adrenal glands were significantly (p < 0.01) lower in the Lou/C rats than in the WAL rats and the two other groups, respectively. DISCUSSION: The ability of the Lou/C rats to accumulate less body fat than their equally food restricted Wistar counterparts (WFR) suggests a difference in basal metabolism in this strain of rats that resembles obesity-resistant rats. PMID- 11886944 TI - Mind and matter: OEM and the World Trade Center. PMID- 11886945 TI - Chronic effects of air pollution. PMID- 11886947 TI - Temporal and spatial relations between age specific mortality and ambient air quality in the United States: regression results for counties, 1960-97. AB - OBJECTIVE: To investigate longitudinal and spatial relations between air pollution and age specific mortality for United States counties (except Alaska) from 1960 to the end of 1997. METHODS: Cross sectional regressions for five specific periods using published data on mortality, air quality, demography, climate, socioeconomic status, lifestyle, and diet. Outcome measures are statistical relations between air quality and county mortalities by age group for all causes of death, other than AIDS and trauma. RESULTS: A specific regression model was developed for each period and age group, using variables that were significant (p<0.05), not substantially collinear (variance inflation factor <2), and had the expected algebraic sign. Models were initially developed without the air pollution variables, which varied in spatial coverage. Residuals were then regressed in turn against current and previous air quality, and dose-response plots were constructed. The validity of this two stage procedure was shown by comparing a subset of results with those obtained with single stage models that included air quality (correlation=0.88). On the basis of attributable risks computed for overall mean concentrations, the strongest associations were found in the earlier periods, with attributable risks usually less than 5%. Stronger relations were found when mortality and air quality were measured in the same period and when the locations considered were limited to those of previous cohort studies (for PM(2.5) and SO(4)(2-)). Thresholds were suggested at 100-130 microg/m(3) for mean total suspended particulate (TSP), 7-10 microg/m(3) for mean sulfate, 10-15 ppm for peak (95th percentile) CO, 20-40 ppb for mean SO(2.) Contrary to expectations, associations were often stronger for the younger age groups (<65 y). Responses to PM, CO, and SO(2) declined over time; responses in elderly people to peak O(3) increased over time as did responses to NO(2) for the younger age groups. These results generally agreed with previous prospective cohort and ecological studies for comparable periods, age groups, and pollutants, but they also suggest that the results of those previous studies may no longer be applicable. CONCLUSIONS: Spatially derived relations between air quality and mortality vary significantly by age group and period and may be sensitive to the locations included in the analysis. PMID- 11886946 TI - Review of recent epidemiological studies on paternal occupations and birth defects. AB - The main findings reported by recent epidemiological studies on paternal occupations and birth defects are reviewed, and the main limitations associated with these studies discussed. Epidemiological studies on paternal occupations and birth defects were reviewed for the period 1989 to 1999 inclusive. Systematic searches were made with search engines with related keywords. There were several common paternal occupations that were repeatedly reported to be associated with birth defects. These paternal occupations were janitors, painters, printers, and occupations exposed to solvents; fire fighters or firemen; and occupations related to agriculture. The common weaknesses in most of these studies include inaccurate assessment of exposures, different classification systems, different inclusion criteria of birth defects, and low statistical power. It is concluded that epidemiological studies, reported in the past decade, suggest that several common paternal occupations are associated with birth defects. Future studies could be focused on these specific, rather than general, occupational groups so that causative agents may be confirmed and thus enable appropriate preventive measures to be taken. PMID- 11886948 TI - Mental health insurance claims among spouses of frequent business travellers. AB - OBJECTIVES: Following up on two earlier publications showing increased psychological stress and psychosocial effects of travel on the business travellers this study investigated the health of spouses of business travellers. METHODS: Medical claims of spouses of Washington DC World Bank staff participating in the medical insurance programme in 1997-8 were reviewed. Only the first of each diagnosis with the ninth revision of the international classification of diseases (ICD-9) recorded for each person was included in this analysis. The claims were grouped into 28 diagnostic categories and subcategories. RESULTS: There were almost twice as many women as men among the 4630 identified spouses. Overall, male and female spouses of travellers filed claims for medical treatment at about a 16% higher rate than spouses of non travellers. As hypothesised, a higher rate for psychological treatment was found in the spouses of international business travellers compared with non-travellers (men standardised rate ratios (RR)=1.55; women RR=1.37). For stress related psychological disorders the rates tripled for both female and male spouses of frequent travellers (>or= four missions/year) compared with those of non travelling employees. An increased rate of claims among spouses of travellers versus non-travellers was also found for treatment for certain other diagnostic groups. Of these, diseases of the skin (men RR=2.93; women RR=1.41) and intestinal diseases (men RR=1.31; women RR=1.47) may have some association with the spouses' travel, whereas others, such as malignant neoplasms (men RR=1.97; women RR=0.79) are less likely to have such a relation. CONCLUSION: The previously identified pattern of increased psychological disorders among business travellers is mirrored among their spouses. This finding underscores the permeable boundary between family relations and working life which earlier studies suggested, and it emphasises the need for concern within institutions and strategies for prevention. PMID- 11886949 TI - Work environment and neck and shoulder pain: the influence of exposure time. Results from a population based case-control study. AB - OBJECTIVES: To study associations between long term and short term exposure to different work environmental conditions and the incidence of neck or shoulder pain. The results were obtained as part of the MUSIC-Norrtalje study, which is a population based case-control study conducted in Sweden in 1993-7. METHODS: The cases were people from the study base who sought medical care or treatment for neck or shoulder pain. Information on physical and psychosocial conditions in the work environment, currently and 5 years ago, and lifestyle factors, was obtained by self administered questionnaires from 310 cases and 1277 randomly selected referents. RESULTS: Associations between both physical and psychosocial exposures in the work environment and seeking care for neck or shoulder pain were found. The risk patterns differed for the sexes, and risk ratios exceeding 1.5 were more often found among women than among men. Generally, subjects who had experienced a recent increase of exposure were more likely (relative risk (RR) 2.1-3.7) to seek care than those who had been exposed long term (RR 1.5-1.8). Among women, an increased amount of visual display terminal (VDT) work, work above shoulder level, and reduced opportunities to acquire new knowledge, and among men, an increased amount of seated work were associated with neck or shoulder pain. This might indicate short induction periods for neck or shoulder pain for these exposures. However, for repetitive work with the hands and hindrance at work among women, and possibly also local vibrations among men, the induction periods seem to be longer. Interactive effects between factors, both at work and in the family, were found, but only among women. CONCLUSIONS: Associations between some exposures in the work environment and seeking care for neck or shoulder pain were found. The high RRs for short term exposure might indicate that for many factors the induction period for neck or shoulder pain is short. PMID- 11886950 TI - Detection of workers sensitised to high molecular weight allergens: a diagnostic study in laboratory animal workers. AB - OBJECTIVES: To determine whether results from questionnaires, exposure measurements, and laboratory tests, commonly used in occupational health practice, can predict the presence or absence of sensitisation in workers exposed to high molecular weight (HMW) allergens. The study aims to develop and validate a diagnostic rule to predict sensitisation in laboratory animal workers. The main reason for such research is efficiency. METHODS: Baseline data from 551 laboratory animal workers participating in an ongoing cohort study, bridging a period of 3 years, were used for diagnostic research. Data from 472 workers participating in the study during the first period were used to develop a prediction rule; these workers represented the derivation set. Data from 79 workers, participating during the second period, were used to evaluate the rule's performance--the validation set. Serum samples were analysed for specific immunoglobulin E (IgE) antibodies against common and laboratory animal allergens. Questionnaire items, exposure determinants, IgE serology, skin prick tests (SPTs), and lung function tests were analysed, corresponding to diagnostic investigation, in a multiple logistic regression model. The accuracy of the model was evaluated by receiver operating characteristic (ROC) curve analysis, and by comparison of the predicted and observed prevalences. RESULTS: Asthmatic symptoms, (work related) allergic symptoms, sex, occupational exposure to rats, and a positive SPT to common allergens, showed the best performance in discriminating workers at high or at low risk of being sensitised. CONCLUSION: High and low risk categories for work related sensitisation can be distinguished from simple questionnaire data and SPT results. The method can easily be applied in occupational medical practice and may markedly increase the efficiency of occupational health surveillance in laboratory animal workers as well as other workers exposed to HMW allergens. PMID- 11886951 TI - Reported chemical sensitivities in a health survey of United Kingdom military personnel. AB - OBJECTIVE: To report the prevalence of self reported chemical sensitivities in three cohorts of United Kingdom service personnel. METHOD: Cross sectional postal survey of three cohorts of United Kingdom military personnel comprising Gulf veterans (n=3531), those who had served in Bosnia (n=2050), and those serving during the Gulf war but not deployed there (Era cohort, n=2614). RESULTS: Sensitivity to at least one everyday chemical was reported by a considerable proportion of all three cohorts, and particularly by veterans of the Gulf war (Era: 14%; Bosnia: 13%; Gulf: 28%). CONCLUSION: Reported chemical sensitivities were common in all three military cohorts. Our understanding of chemical sensitivities remains limited and objective evidence for a causal link between low level exposures to chemicals and reported symptoms is lacking. Given their frequency in the population, further work in this area is necessary. PMID- 11886952 TI - Suicide mortality among electricians in the Swedish construction industry. AB - OBJECTIVES: To investigate the risk of suicide in Swedish electricians employed in the construction industry. A few studies have indicated an increased risk of suicide for electricians in the construction industry and electricians exposed to electromagnetic fields. METHODS: This is a cohort study. Electricians were identified through a computerised register of construction workers who had participated in health examinations in 1971-92. In this register, 33,719 male electricians were identified together with a reference group consisting of 72,653 male glass or woodworkers. Through a linkage with the Swedish Death Register, the cause of death was identified to the end of 1997. Mortality as a result of suicide was also compared with the general population with adjustments for sex, age, and period. RESULTS: The risk of mortality from suicide was decreased for electricians (standardised mortality ratio (SMR) 0.58, 95% confidence interval (95% CI) 0.47 to 0.71) and for the reference group of construction workers (SMR 0.81, 95% CI 0.72 to 0.91) compared with the general population. CONCLUSION: Contrary to some other studies, risk of suicide was not increased among electricians in the construction industry. PMID- 11886953 TI - Sickness absence in doctors. PMID- 11886954 TI - Offspring sex ratios of people exposed to dioxin and dioxin-like compounds. PMID- 11886956 TI - Epidemiology of disability in rheumatoid arthritis. PMID- 11886958 TI - Alcohol intake in rheumatic disease: good or bad? PMID- 11886955 TI - Translating evidence about occupational conditions into strategies for prevention. PMID- 11886959 TI - Silicone breast implants: correlation between implant ruptures, magnetic resonance spectroscopically estimated silicone presence in the liver, antibody status and clinical symptoms. AB - OBJECTIVE: To determine the impact of implant integrity on clinical symptoms and antibody status in women with silicone breast implants (SBIs). METHODS: Ninety consecutive women were examined by means of magnetic resonance imaging (MRI) to assess the integrity of their silicone breast implants. The presence of silicone in the liver was estimated by (1)H localized stimulated echo acquisition mode (STEAM) magnetic resonance spectroscopy (MRS). Results were correlated with patients' complaints, as evaluated by a standardized questionnaire, physical examination by a rheumatologist and antibody screening. RESULTS: Breast MRI revealed defects in 24 patients (26.6%); in 13 (54.2%) of these women, silicone was detected in the liver by MRS. Of the 66 patients with MRI-estimated intact implants, 15 (22.7%) had apparent silicone in their liver, arguing for gel bleeding. Clinically, two patients had had rheumatoid arthritis before SBIs, whereas the other patients revealed no typical symptoms of arthritis or connective tissue disease (CTD). The patients with MRS evidence of silicone in the liver had no statistically significant differences in their complaints with the exception of the most frequent symptom, tingling/numbness of the fingers (82.1 vs 51.6%, P=0.006). A positive pattern of antinuclear antibodies (ANA) was obtained in 13 of the 28 MRS-positive patients (46.4%) and in 15 of the 62 MRS negative patients (24.2%, P=0.033). However, in only one of these 28 ANA-positive patients was a specific weak antibody titre against SS-A detected by ELISA. CONCLUSION: Implant integrity has no major impact on rheumatic symptoms of women with SBIs. This finding supports the standpoint that silicone does not cause either a specific CTD or any other distinct disease entity. However, the moderately increased incidences of ANA-positivity and neuropathy-associated symptoms require explanation. PMID- 11886960 TI - Plasminogen activator inhibitor type-1 deficiency attenuates murine antigen induced arthritis. AB - OBJECTIVE: To examine the role of plasminogen activator inhibitor type-1 (PAI-1), the major fibrinolytic inhibitor, in vivo during murine antigen-induced arthritis (AIA). METHODS: AIA was induced in PAI-1-deficient mice and control wild-type mice. Arthritis severity was evaluated by technetium 99m (99mTc) uptake in the knee joints and by histological scoring. Intra-articular fibrin deposition was examined by immunohistochemistry and synovial fibrinolysis quantitated by tissue D-dimer measurements and zymograms. RESULTS: Joint inflammation, quantitated by 99mTc uptake, was significantly reduced in PAI-1(-/-) mice on day 7 after arthritis onset (P<0.01). Likewise, synovial inflammation, evaluated by histological scoring, was significantly decreased in PAI-1-deficient mice on day 10 after arthritis onset (P<0.001). Articular cartilage damage was significantly decreased in PAI-1(-/-) mice, as shown by histological grading of safranin-O staining on day 10 after arthritis onset (P<0.005). Significantly decreased synovial accumulation of fibrin was observed by day 10 in arthritic joints of PAI 1(-/-) mice (P<0.005). Accordingly, the synovial tissue content of D-dimers, the specific fibrin degradation products generated by plasmin, were increased in PAI 1(-/-) mice (P<0.02). Finally, as expected, PA activity was increased in synovial tissues from PAI-1(-/-) mice, as shown by zymographic analysis. CONCLUSIONS: These results indicate that deficiency of PAI-1 results in increased synovial fibrinolysis, leading to reduced fibrin accumulation in arthritic joints and reduced severity of AIA. PMID- 11886961 TI - Relevant change in radiological progression in patients with hip osteoarthritis. I. Determination using predictive validity for total hip arthroplasty. AB - OBJECTIVE: To determine a cut-off point above which a change in joint space width (JSW) could be considered as relevant in patients with hip osteoarthritis (OA) on the basis of predicted need for subsequent total hip arthroplasty (THA). METHODS: A multicentre, prospective, longitudinal, 5-yr follow-up study was performed. A pelvic radiograph was obtained at entry and after 1 and 2 yr. For each film, the narrowest JSW was measured using a 0.1 mm graduated magnifying glass. The absolute and relative differences between baseline and 1 and 2 yr of follow-up were calculated. We determined the cut-off points above which an absolute or relative decrease in JSW between baseline and 1 and 2 yr of follow-up could be considered relevant on the basis of the predicted need for THA during the remaining years of the study. The need for THA was categorized as 'yes' or 'no'. Thereafter, for each observed change in JSW (0.1 per 0.1 mm or 1% per 1%), the sensitivity and specificity for subsequent THA were calculated. The choice of cut off was based on maximal sensitivity and specificity, using the graphic representation of correct classification probabilities. In this way it was possible to obtain the best measured JSW threshold with maximal true positive and minimal false positive results. RESULTS: A total of 423 and 385 patients met the criteria for analysis using the decrease in JSW between baseline and 1 and 2 yr respectively. The best cut-off points were absolute decreases in JSW of 0.2 and 0.4 mm and relative decreases in JSW of 15 and 20% after 1 and 2 yr respectively, with corresponding ranges of sensitivity and specificity of 68-75 and 67-78%. CONCLUSION: This work determined the cut-off above which a change in JSW could be considered clinically relevant in patients with hip OA, on the basis of predicted subsequent need for THA. For validation, similar studies should be conducted in other countries with different health-care systems. PMID- 11886962 TI - Relevant change in radiological progression in patients with hip osteoarthritis. II. Determination using an expert opinion approach. AB - AIM: To determine the minimum clinically important difference (MCID) in joint space width (JSW) progression in patients with hip osteoarthritis (OA), based upon evaluation by a panel of clinical experts as a gold standard. METHODS: A sample of 298 patients with hip OA was selected from a multicentre, prospective, longitudinal, 3-yr follow-up study. A pelvic radiograph was obtained at entry and after 3 yr. For each film, the narrowest JSW was measured using a 0.1-mm graduated magnifying glass. The difference between baseline and 3-yr follow-up JSW was calculated. Two senior rheumatologists, who were experts in osteoarthritis, evaluated each pair of films and noted whether a clinically relevant deterioration in osteoarthritis stage occurred at 3 yr compared with baseline. Interobserver reliabilities were evaluated using the kappa coefficient and proportions of agreements. Then, for each measured difference in JSW (0.1 mm per 0.1 mm), the sensitivity and specificity for MCID, defined as the assessment of expert 1, expert 2 or a combination of both, were calculated. This allowed us to obtain, from graphic representations of the correct classification probabilities, the best measured JSW threshold, with the maximal true positive and the minimal false positive results. RESULTS: The mean measured change in JSW was -0.63 +/- 0.74 mm. Experts 1 and 2 considered the decrease in JSW to be clinically relevant in 122 (40.9%) and 100 pairs (33.6%) respectively. The proportion of agreements between the experts was 79.9%, with a kappa coefficient of 0.572. The best measured JSW threshold was -0.4 mm for expert 1, expert 2 and the combination of both; sensitivity and specificity were 0.75 and 0.8, 0.71 and 0.72, and 0.75 and 0.7 respectively. CONCLUSION: This study suggests that a change of at least 0.4 mm in the radiological JSW could be considered clinically relevant. Other studies using other sets of patients and other methods are needed for validation. PMID- 11886963 TI - The presence of the HLA-DRB1 shared epitope correlates with erosive disease in Chilean patients with rheumatoid arthritis. AB - OBJECTIVE: To assess the contribution of the HLA-DRB1 shared epitope (SE) to the radiological outcome of rheumatoid arthritis (RA) after 6 yr of follow-up in a reported series of 129 Chilean patients with established disease. METHODS: A prospective study was conducted between 1992 and 1998 using hand radiographs to assess disease outcome in a published series of patients in whom two doses of the SE were present in 20%, one dose was present in 34% and the SE was absent in 46%. At study entry, 29 of the 92 patients with hand radiographs were at Steinbrocker stages I or II (non-erosive), with a median disease duration of 2.8 yr (0.4-17). RESULTS: In 1998, 113 (87%) of the patients were alive. One hundred and eight patients underwent complete clinical evaluation. Their median age was 57 yr (range 30-81) and the median disease duration was 15 yr (6-50). We were able to study 25 of the 29 patients who had non-erosive disease at study entry in 1992. We found that 10 of 11 patients having one or two doses of the SE developed erosive disease compared with three of 14 without the SE (Yates' corrected P=0.0023, relative risk 4.24, 95% confidence interval 1.53-11.77). CONCLUSIONS: These observations support and extend the notion that the presence of the SE in one or two doses can predict the development of erosions even in RA populations in whom the SE is not as prevalent as in Caucasians. PMID- 11886964 TI - Exercise can reverse quadriceps sensorimotor dysfunction that is associated with rheumatoid arthritis without exacerbating disease activity. AB - OBJECTIVES: To compare quadriceps sensorimotor function, lower limb functional performance and disability in patients with rheumatoid arthritis (RA) and healthy subjects, and to investigate the efficacy and safety of a brief rehabilitation regime. METHODS: Quadriceps strength, voluntary activation, proprioceptive acuity and the aggregate time [aggregate functional performance time (AFPT)] taken to perform four common activities [aggregate functional performance time (AFPT)] were compared between 103 RA patients who had lower limb involvement and 25 healthy subjects. In addition, disability (Health Assessment Questionnaire), clinical disease activity and the plasma concentration of proinflammatory cytokines were measured in the RA patients. In a follow-on randomized controlled trial of rehabilitation, these variables were used as baseline data for 93 of the RA patients, who were randomized to a rehabilitation or a control group. Changes in the variables were analysed within and between groups. RESULTS: Compared with healthy subjects, RA patients had weaker quadriceps [mean difference 157 N; 95% confidence interval (CI) 125-189], poorer activation (8%, 95% CI 4.5-15) and proprioceptive acuity (0.8 degrees, 95% CI 0.4-1.3) and took longer to perform the AFPT (34 s, CI 23.5-44.8). Rehabilitation increased quadriceps strength (mean increase 61 N, 95% CI 28-95) and voluntary activation (8%, 95% CI 3-12.4) and decreased the AFPT (12.3 s, 95% CI -2 to 27.7) and subjective disability (0.21 HAQ points, 95% CI 0-0.35) without exacerbating disease activity. All the improvements were maintained at the 6-month follow-up. There was no change during the control period. CONCLUSIONS: Patients with RA that affected their lower limb had quadriceps sensorimotor deficits that were associated with lower limb disability. A clinically applicable rehabilitation regime increased quadriceps sensorimotor function and decreased lower limb disability without exacerbating pain or disease activity. For patients with well-controlled RA that causes lower limb involvement, the regime is effective and safe. PMID- 11886965 TI - Health-care use by rheumatoid arthritis patients compared with non-arthritic subjects. AB - OBJECTIVE: To compare the use of health-care by rheumatoid arthritis (RA) patients and non-arthritic subjects (NA) and to look for factors determining their patterns of health-care use. METHODS: A multicentre cohort of 223 RA and 446 NA subjects matched for age, gender, period of data collection and residence were questioned about their use of health-care services. Patterns of health-care use were identified by principal components analysis. Factors determining the use of health-care services were assessed by multiple linear and logistic regression analysis. RESULTS: The proportions of RA subjects who declared having had at least one contact with the health-care system in the previous 12 months and in the previous 4 weeks were higher than those for NA subjects for all health and social professionals except dentists and homeopaths. Types of health-care use explored were hospital, prescribed, general ambulatory and specialized ambulatory care. Factors determining health-care use were disease status, administrative area, employment status and age. CONCLUSIONS: RA subjects use health-care services more widely than NA subjects. Variation in recourse behaviour is related to differences within administrative areas. PMID- 11886966 TI - HFE mutations in an inflammatory arthritis population. AB - OBJECTIVES: To determine the value of screening patients with inflammatory arthritis for haemochromatosis-associated mutations in the HFE gene. METHODS: We screened 1000 patients with inflammatory arthritis and 1000 controls for the HFE gene mutations that are associated with haemochromatosis. The arthritis patients were diagnosed between 1989 and 1995 and their blood DNA was archived as part of the Norfolk Arthritis Register project. RESULTS: Five out of 1000 (0.005) patients in the arthritis group were homozygous for the HFE C282Y mutation. This frequency is the same as the frequency of 5/1000 (0.005) for C282Y homozygosity observed in the normal population. It is slightly above the predicted frequency of homozygosity of 0.0044 derived from the gene frequency in the normal population. CONCLUSIONS: These data suggest that most of the C282Y homozygotes occurred in this arthritis group by chance and that their arthritis was incidental to their HFE genotype. This implies that screening for HFE mutations among patients with inflammatory arthritis would infrequently identify patients whose arthritis might benefit from additional treatment. PMID- 11886968 TI - Psychological distress and changes in the activity of systemic lupus erythematosus. AB - OBJECTIVE: To determine if changes in depressive symptoms or anxiety lead to changes in the activity of systemic lupus erythematosus (SLE). METHODS: Twenty three patients with SLE were examined prospectively every 2 weeks for up to 40 weeks. At each assessment, patients completed the Centers for Epidemiologic Studies--Depression scale (CES-D), the State subscale of the State-Trait Anxiety Inventory and a global assessment of the activity of their SLE by visual analogue scale. SLE activity was also assessed at each visit by physician global assessment, the Systemic Lupus Activity Measure (SLAM), the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) and the European Consensus Lupus Activity Measure (ECLAM). RESULTS: Changes in depression and anxiety were positively correlated with simultaneous changes in the patient global assessment of SLE activity and in the SLAM, but not with changes in the physician global assessment, SLEDAI or ECLAM. Depression and anxiety scores were also correlated with patient global assessments and SLAM scores 2 weeks later, but lagged scores were not significantly associated with the patient global assessment or SLAM after controlling for current depression and anxiety scores. The associations between depression and anxiety scores and the SLAM were not present when SLAM scores were modified to exclude ratings of depression and fatigue. No measure of SLE activity increased in the 2 weeks immediately after a large increase in CES-D or State Anxiety scores. CONCLUSIONS: Depression and anxiety scores parallel changes in patients' assessments of the activity of their SLE. We found no evidence to support the hypothesis that psychological distress causes increased SLE activity. PMID- 11886967 TI - An association between the CTLA4 exon 1 polymorphism and early rheumatoid arthritis with autoimmune endocrinopathies. AB - OBJECTIVE: To examine the allelic association of the single nucleotide polymorphism (CTLA4A/G) in exon 1 of the cytotoxic T lymphocyte antigen-4 (CTLA4) gene with early rheumatoid arthritis (RA). METHODS: One hundred and twenty-three unrelated white probands with early RA from the north-east of England and 349 local ethnically matched controls were studied. The CTLA4A/G polymorphism was genotyped with a polymerase chain reaction (PCR) method and digestion with the restriction enzyme Bst71I. Probands were also screened by allele-specific PCR for alleles HLA DRB1*01 and DRB1*04. RESULTS: The frequency of the G allele at CTLA4A/G was significantly increased in probands with early RA compared with controls [43 vs 36%; P=0.028, odds ratio (OR) 1.35, 95% confidence interval (CI) 1.01-1.82]. Most of this increased frequency was attributable to RA individuals with coexisting autoimmune thyroid disease or type 1 diabetes (58 vs 36% in controls; P=0.005, OR 2.50, CI 1.29-4.84). The frequency of the G allele in RA patients without autoimmune endocrinopathy was 40%, which was not significantly different from that in controls (P=0.140). CONCLUSION: The association between the CTLA4 G allele and early RA is largely explained by individuals with RA who have coexisting autoimmune endocrinopathies. PMID- 11886969 TI - The mechanical joint score: a new clinical index of joint damage in rheumatoid arthritis. AB - OBJECTIVES: To evaluate the mechanical joint score (MJS) in terms of its reliability between observers and over time, its ease of use and its relationship with conventional measures of rheumatoid arthritis (RA) disease activity, severity and functional outcome. METHODS: The MJS was evaluated in 103 patients with reference to the following joints: total proximal interphalangeal (PIP) joints, total metacarpophalangeal (MCP) joints, wrists, elbows, shoulders, hips, knees, ankles and total metatarsophalangeal (MTP) joints. The score was based on the appearance of the joints on a scale of 0-3, 0 representing no abnormality and 3 severe abnormality or previous surgery. The MJS was evaluated in terms of its intra- and inter-observer variability and its content, construct and criterion validities. A subset of 29 patients were re-evaluated after 5 yr to examine change in MJS over time. RESULTS: The MJS performed well in terms of inter observer and intra-observer reliability. The MJS showed strong correlation with the Larsen X-ray score of hands and feet (Spearman correlation coefficient 0.74) and with the modified Health Assessment Questionnaire (Spearman correlation coefficient 0.56) and only weak correlation with indices of disease activity, such as the Ritchie index and erythrocyte sedimentation rate. The MJS showed highly significant positive change over time. CONCLUSION: The MJS is a reliable clinical index of joint damage and may be a useful new outcome measure in RA. PMID- 11886971 TI - Acupuncture in chronic epicondylitis: a randomized controlled trial. AB - OBJECTIVE: To evaluate the clinical efficacy of acupuncture in the treatment of chronic lateral epicondylitis. METHODS: In a randomized, investigator- and patient-blinded, controlled clinical study, 23 patients were treated with real acupuncture and 22 patients received sham acupuncture. Patients each received 10 treatments, with two treatments per week. The primary outcome variables were maximal strength, pain intensity (verbal rating scale) and disability scale (Disabilities of the Arm, Shoulder and Hand questionnaire). Patients were examined at baseline (1 week before the start of treatment) and at follow-up 2 weeks and 2 months after the end of treatment. RESULTS: There was no significant difference between the groups at baseline for any outcome parameter. Two weeks and 2 months after the end of treatment, there were significant reductions in pain intensity and improvements in the function of the arm and in maximal strength in both treatment groups. At the 2-week follow-up these differences were significantly greater for all outcome parameters in the group treated with real acupuncture. At 2 months the function of the arm was still better in this group than in the sham acupuncture group; however, the differences in pain intensity and maximal strength between the groups were no longer significant. CONCLUSION: In the treatment of chronic epicondylopathia lateralis humeri, acupuncture in which real acupuncture points were selected and stimulated was superior to non specific acupuncture with respect to reduction in pain and improvement in the functioning of the arm. These changes are particularly marked at early follow-up. PMID- 11886970 TI - Radiographic outcome after three years of patients with early erosive rheumatoid arthritis treated with intramuscular methotrexate or parenteral gold. Extension of a one-year double-blind study in 174 patients. AB - OBJECTIVE: To compare the radiographic outcomes after 36 months in patients with early erosive rheumatoid arthritis (RA) who were treated with methotrexate (MTX) or gold sodium thiomalate (GSTM). METHODS: In a randomized, double-blind fashion, 174 patients from two centres were assigned to receive weekly intramuscular injections of either 15 mg MTX or 50 mg GSTM. After 12 months, the study was continued as an open prospective study for an additional 2 yr, administering the same amount of MTX and half of the GSTM dose. Radiographic outcomes were assessed by standardized methods in all patients at baseline and after 6, 12, 24 and 36 months. RESULTS: Intention-to-treat analysis showed that patients treated with MTX had higher radiographic scores and more erosive joints at all follow-up points. However, there was no statistically significant difference between the two treatment groups. The progression rate was significantly slower during the second and third years of follow-up in both groups. Baseline and time-integrated (area under the curve over 6 months) disease activity parameters were good predictors of radiographic outcome after 3 yr. Seropositivity was not an independent predictor of progression. However, patients who were positive for rheumatoid factor had higher time-integrated disease activity (with less response to treatment) and thus their disease was significantly more progressive. CONCLUSION: Both of the disease-modifying compounds used in this study, MTX and GSTM, were able to reduce the slope of radiographic progression during 3 yr of follow-up. There was some advantage for parenteral gold but no significant intergroup difference. PMID- 11886972 TI - Survey of arthroscopy performed by rheumatologists. AB - OBJECTIVE: To determine the international distribution and practice of arthroscopy performed by rheumatologists and to evaluate proposed guidelines on minimum standards for training in arthroscopy in the context of current clinical practice. METHODS: A questionnaire was sent to all rheumatology centres identified as practising arthroscopy, by (i) searching Medline from 1966 to 1999, (ii) searching the abstract books of the annual general meetings of ACR, BSR and EULAR from 1980 to 1999, and (iii) correspondence with all the centres identified. RESULTS: Thirty-six rheumatology centres were confirmed as performing arthroscopy (24 in Europe, 10 in USA and two in Australia) and 33 (92%) centres completed the questionnaire. Twenty-five (76%) of the 33 centres performing arthroscopy had started to perform it since 1990 and 72 rheumatologists are now trained in arthroscopy. A total of 16532 arthroscopies had been performed (median=220 arthroscopies/centre, range 20-5000); 50.5% of the arthroscopies had a primary clinical indication and 49.5% had a primary research indication. Most centres fulfilled the minimum standards for arthroscopic facilities and the proposed minimum standards in training were acceptable to 76% of respondents. Complication rates were calculated for 15682 arthroscopies where routine follow up data were available [joint infection, 16 (0.1%); wound infection, 17 (0.1%); haemarthrosis, 141 (0.9%); deep venous thrombosis, 31 (0.2%); neurological damage, 3 (0.02%), thrombophlebitis, 12 (0.08%), other, 8 (0.06%)]. Irrigation volume correlated with wound infection rate (r=0.41, P=0.03) and centres performing cartilage biopsy had a higher rate of haemarthrosis (P=0.007). CONCLUSION: The last decade has seen rapid growth in arthroscopy performed by rheumatologists in an out-patient setting under local and regional anaesthesia. Proposed minimum standards for training in rheumatological arthroscopy reflect current practice accurately and are acceptable to the majority of arthroscopists. Complication rates of rheumatological arthroscopy are similar to those reported in the orthopaedic literature. PMID- 11886973 TI - Miller Fisher syndrome in adult onset Still's disease: case report and review of the literature of other neurological manifestations. AB - Adult-onset Still's disease (AOSD) is a multi-system inflammatory disorder characterized by high spiking fevers, evanescent salmon-coloured rash, arthralgias or arthritis, hepatosplenomegaly, lymphadenopathy and sore throat. There is no specific test or combination of tests that can establish the diagnosis of AOSD and patients may present with other systemic involvement including neurological manifestations in 7-12% of cases. We present a complex case of a patient with AOSD who developed the Miller-Fisher variant of Guillain Barre syndrome. This immunological disorder of the nervous system has not been described in association with AOSD before. We also review the literature on other neurological manifestations in AOSD. AOSD mimics different disease processes and its multi-system manifestations may complicate the picture further. PMID- 11886974 TI - Tumour necrosis factor alpha G-->A -238 and G-->A -308 polymorphisms in juvenile idiopathic arthritis. AB - OBJECTIVES: To study G-->A -238 and G-->A -308 polymorphisms in the promoter region of the tumour necrosis factor (TNF) alpha gene in patients with juvenile idiopathic arthritis (JIA). We analysed whether there were any associations between these polymorphisms and the type of JIA and/or the clinical course of the disease in two populations. METHODS: The first group consisted of 51 Turkish JIA patients and the second consisted of 159 JIA patients from the Czech Republic. Healthy individuals (93 and 100) from each country served as controls. Subgroups of JIA were defined according to the Durban criteria. The course of the disease was defined on the basis of the physician's global evaluation of disease activity, the swollen and tender joint count and the erythrocyte sedimentation rate. RESULTS: In both JIA cohorts, the distribution of genotypes was not significantly different among the types of JIA. The G-->A -238 polymorphism did not have an effect on the patients' outcome in either group. The G-->A -308 polymorphism was significantly associated with a poor outcome in the Turkish group (P=0.005) but there was no association in the Czech patients. Some features of JIA in Turkish patients differed from those in Czech patients. CONCLUSIONS: Genetic differences may accompany the phenotypic differences found in the Turkish group. Although larger numbers of patients are clearly needed to verify this, we suggest that the G-->A -308 polymorphism may be operative in defining disease outcome in selected groups. PMID- 11886975 TI - Etanercept in four children with therapy-resistant systemic juvenile idiopathic arthritis. PMID- 11886976 TI - Foreign body synovitis induced by a crown-of-thorns starfish. PMID- 11886977 TI - Pamidronate: a novel treatment for the SAPHO syndrome? PMID- 11886978 TI - Miliary tuberculosis after biological therapy for rheumatoid arthritis. PMID- 11886979 TI - Methotrexate-induced nephrogenic diabetes insipidus: first case report. PMID- 11886980 TI - Successful treatment of refractory mononeuritis multiplex secondary to rheumatoid arthritis with the anti-tumour necrosis factor alpha monoclonal antibody infliximab. PMID- 11886981 TI - The London Rheumatology Club 1977-2000. PMID- 11886982 TI - Pyoderma gangrenosum associated with Wegener's granulomatosis: partial response to mycophenolate mofetil. PMID- 11886983 TI - Robin Goodfellow. PMID- 11886985 TI - The craniofacial surgeon as amateur geneticist. AB - Craniofacial surgeons, by nature and training, focus on how to correct anomalies rather than on why they occur. Surgeons often leave diagnosis and etiopathogenic speculation to geneticists. Craniofacial surgeons should cross over the specialty line and learn to think like geneticists. This article reviews definitions of basic words in the genetic language and emphasizes the three diagnostic levels, phenotypic, pathogenic, and genetic, for the principal categories of craniofacial anomalies. Whenever possible, examples are given to illustrate how genetic knowledge can influence surgical strategy. As a member of the perinatal team, the craniofacial surgeon must be "cyber-savvy" to counsel parents and communicate with geneticists. PMID- 11886986 TI - International Task Force on Volunteer Cleft Missions. AB - The International Task Force on Volunteer Cleft Missions was set up to provide a report to be presented at the Eighth International Congress of Cleft Palate and Associated Craniofacial Anomalies on September 12, 1997, in Singapore. The aim of the report was to provide data from a wide range of different international teams performing volunteer cleft missions and, thereafter, based on the collected data, to identify common goals and aims of such missions. Thirteen different groups actively participating in volunteer cleft missions worldwide were selected from the International Confederation of Plastic and Reconstructive Surgery's list of teams actively participating in volunteer cleft missions. Because of the time frame within which the committee had to work, three groups that did not respond by the stipulated deadline were omitted from the committee. The represented members and their respective institutions have undertaken more than 50 volunteer cleft missions to underdeveloped nations worldwide within the last 3 years. They have visited over 20 different countries, treating more than 3,500 patients worldwide. Based on the data collected and by consensus, the committee outlined recommendations for future volunteer cleft missions based on 1) mission objectives, 2) organization, 3) personal health and liability, 4) funding, 5) trainees in volunteer cleft missions, and 6) public relations. The task force believed that all volunteer cleft missions should have well-defined objectives, preferably with long-term plans. The task force also decided that it was impossible to achieve a successful mission without good organization and close coordination. All efforts should be made, and care taken, to ensure that there is minimal morbidity and no mortality. Finally, as ambassadors of goodwill and humanitarian aid, the participants must make every effort to understand and respect local customs and protocol. The main aims are to provide top-quality surgical service, train local doctors and staff, develop and nurture fledgling cleft programs, and, finally, make new friends. PMID- 11886987 TI - Clinicopathological evaluation of parotid gland tumors: a retrospective study. AB - Two hundred and thirty sequential parotid tumors seen from March 1985 to 1995 were reviewed for their clinical presentation, diagnostic evaluation, pathological diagnosis, treatment modalities, and age and sex distribution. An asymptomatic mass was the most common clinical presentation. All of the operations were performed by the same surgical team. Total and superficial parotidectomy was used for the treatment of the lesions and none of the patients underwent limited excision. Retrograde approach in 79 (34.4%) patients and anterograde approach in 151 (65.6%) was used. Eighteen patients with malignant tumors were followed up in cooperation with the radiation oncology clinic. Tumors were classified according to their histopathologic diagnosis. Among 192 (83%) benign and 38 (17%) malignant tumors, the most common benign tumor of parotid gland was pleomorphic adenoma (79.1%) while the most common malignant lesion was adenocystic carcinoma (44.7%). Incidences of pleomorphic adenoma, adenocystic and epidermoid carcinoma were greater in male patients. Complication rates in benign and malignant tumors were presented and statistically significant difference could not be found between anterograde and retrograde approach in terms of facial nerve injury (P > 0.05). PMID- 11886988 TI - A case of a Japanese patient with cleidocranial dysplasia possessing a mutation of CBFA1 gene. AB - Cleidocranial dysplasia (CCD) is an autosomal dominant human bone disease characterized by hypoplastic or aplastic clavicles, wide cranial sutures, supernumerary teeth, short stature, and other skeletal disorders. Recently, various mutations of the core binding factor (CBFA1) gene have been detected in CCD patients. The CBFA1 gene is a member of the runt family of transcription factors. We experienced one Japanese case of CCD with open sutures, hypoplasia of clavicles and brachydactyly, combined with atlant-axis dislocation. We performed the sequence analysis of the CBFA1 gene and detected a missense mutation of R225W in exon 3. PMID- 11886989 TI - Craniofacial impalement injury: a rake in the face. AB - Impalement injuries describe unusual objects and circumstances in which a body part is either partially embedded (one end sticking out) or transected (through and-through) by a foreign material. In either case, the object remains as part of the wound and is highly conspicuous. These injuries are much more common on the trunk and extremities because of their larger surface areas and the relative ease in which the object may penetrate them without lethal consequences. In the face, however, such injuries are more rare because of protective reflexes that either move the face away from the coming object or permit it only to be deflected away without being embedded. In addition, the face presents a much smaller target than the rest of the body and, therefore, is more infrequently impaled. This paper presents a case that illustrates many of the classic features of the craniofacial impalement-type injury. PMID- 11886990 TI - Intraosseous hemangioma of the mandible: a case report and review of the literature. AB - Intraosseous hemangiomas are benign lesions, occurring often in vertebra and skull. These lesions can present in the head and neck region being the next most common site with a predominance of mandible. There are approximately seventy intraosseous hemangiomas of mandible in literature reported to date. We present an intraosseous hemangioma of the symphysis mandible. We performed a block resection of the mass preserving the mandibular integrity with no complications. There were no recurrences in the follow-up period of 24 months. PMID- 11886992 TI - Minimally invasive Le Fort III distraction. AB - Recent applications of distraction osteogenesis to the Le Fort III osteotomy in patients with craniofacial dysostosis have proven promising (1-3). Distraction has allowed the midfacial segment to be brought further forward and maintained in position with greater stability when compared with the standard technique of intraoperative advancement. Because no bone grafts or plates must be placed, access incisions are necessary only for performance of the osteotomy. In an effort to minimize the morbidity of the procedure, we have begun performing the Le Fort III osteotomy without the coronal incision. Instead, the nasofrontal junction is approached using the medial aspect of an upper blepharoplasty incision. A lower eyelid and gingivobuccal sulcus incision are also used to complete the osteotomy. This technique has resulted in a shorter operative time and decreased blood loss when compared with the Le Fort III distraction procedure using the standard coronal incision. PMID- 11886993 TI - Pleomorphic adenoma of the lachrymal gland: presentation of a clinical case of relapse. AB - Pleomorphic adenoma affecting the lachrymal gland requires a well-grounded clinical and therapeutic protocol to avoid the risk of malignant transformation or disease recurrence, which is quite dangerous at this site. The authors present a clinical case of pleomorphic adenoma recurrence affecting the right lachrymal gland in a patient who first underwent a biopsy, with subsequent resection of the neoplasm. The authors also review the clinical and radiological features enabling a differential diagnosis and describe the radical "exenteratio orbitae" surgery for pleomorphic adenoma recurrence 7 years after the first operation, which was performed because of total derangement of the orbital cavity. The patient remains disease-free 5 years after this surgical treatment. Therefore, it turns out that an incisional biopsy carries a higher biological cost for the patient. PMID- 11886994 TI - Morphological variability of inferior alveolar nerve in low-grade craniofacial microsomia. AB - Craniofacial microsomia (CFM) involves asymmetric hypoplasia and dysmorphogenesis of the facial skeleton. Certain aspects of CFM may be treated by surgical osteotomy and distraction osteogenesis (DO). Mandibular osteotomy places the inferior alveolar nerve at risk. The aim of this study was to investigate radiological landmark relationships to the anatomy of the inferior alveolar nerve in CFM. Application of this understanding will aid intraoperative protection of the inferior alveolar nerve. Six subjects with similar presentations of hemifacial microsomia were selected. Three-dimensional reconstruction CT images was used to locate bony structures that held important relationships with the inferior alveolar nerve. Measurements (of the normal and microsomic sides) were made between fixed landmarks: mandibular notch, mandibular foramen, condyle, back of second and third molar tooth, and mental foramen. The unaffected sides acted as controls. The distance between the normal and the microsomic sides from condyle to mandibular foramen was significantly different. The sizes of condyles differed significantly between the normal and microsomic sides. Most of the remainder of the vertical distance of the ramus and most of the horizontal distance of the body were similar. In conclusion, the inferior alveolar nerve should be found in very similar locations on both normal and microsomic sides in low-grade hemifacial microsomic patients. PMID- 11886996 TI - An anthropometric analysis of indices of severity in the unilateral cleft lip. AB - The aim of this study was to analyze the relationships among three key anthropometric parameters in the unilateral cleft lip to determine the correlations, if any, among these indices of severity. Using a standardized anthropometric documentation protocol, preoperative measurements of 125 unilateral cleft lips (103 complete and 22 incomplete) were performed under general anesthesia by a single surgeon at the time of primary lip repair at the age of 3 months. The following key measurements were analyzed statistically: (1) the philtral height difference (PHD) between the cleft and noncleft sides, (2) the nasal floor width difference (NFWD) between the cleft and noncleft sides, and (3) the cleft width (CW). The mean values of all three indices were greater in the complete group versus the incomplete group. These differences were statistically significant. Linear relationships were obtained between NFWD and GAP, between PHD and GAP, and between PHD and NFWD in the complete group. In contrast, the relationships between PHD and GAP, and between PHD and NFWD were nonlinear in the incomplete group. These findings suggest that there was a strong correlation between the transverse and vertical tissue deficiencies in the complete cleft lip. In incomplete clefts, however, this correlation did not exist. In other words, the incomplete cleft lip can be associated with a severely short philtrum even in the presence of a relatively mild transverse tissue deficit. Therefore, it is not necessarily easier to repair an incomplete cleft lip in terms of the correction of the vertical tissue deficiency. PMID- 11886995 TI - Surgical treatment of TMJ ankylosis: our experience (60 cases). AB - Limitation of mouth opening can be caused by bony or fibrous ankylosis of the temporomandibular joint as sequela to trauma, infection, autoimmune disease, or failed surgery. Various procedures have been reported for treatment of temporomandibular joint (TMJ) ankylosis; this article aims to describe the diagnostic protocol and the surgical procedures adopted at the department of Maxillo-Facial Surgery of Rome University "La Sapienza". Between 1980 and 2000, 123 patients affected by TMJ ankylosis came under our observation; 60 of them (25 females and 35 males of 30 years average age) underwent surgery; bilateral TMJ ankylosis was observed in 21 cases, right-sided in 20 cases, left-sided in 19 cases. In 12 cases coronoid processes were involved. Etiopathogenesis was traumatic in 48 cases, septic in 5 cases, auto-immune (RA and seronegative spondyloarthropathies) in 5 cases; after block removing, arthroplasty was performed with pedunculated flap of temporal muscle (10 cases), Silastic material (11 cases), or lyophilized dura mater (2 cases). Simple condylar shaving was used in the remaining 36 cases. All patients under treatment showed a distinctive improvement both in articular functionality and symptoms; secondary surgery was necessary in seven cases due to the onset of articular complications from previous surgery. Silastic removal was necessary in five cases due to its inducement of foreign body granuloma. Follow-up was performed at 12, 24, and 48 months and 5 years postoperatively. In our opinion the gold standard surgery of TMJ ankylosis today is represented by shaving of articular surfaces and subsequent arthroplasty with or without temporal muscle myofascial flap interposition, whereas the use of Silastic as alloplastic material could be associated to an increased persistence of the local symptoms and a higher risk of foreign body granuloma and it may favor ankylosis relapse and hinder rehabilitation. PMID- 11886997 TI - Color Doppler ultrasound for monitoring free flaps in the head and neck region. AB - The use of microvascular free flaps has become established as a very reliable reconstructive technique following tumor surgery or trauma in the head and neck region. Occasionally, flap compromise may occur which will require immediate re exploration. Early diagnosis of vascular insufficiency is essential. Clinical signs are not always reliable. Numerous systems have been described for monitoring the viability of microsurgical free flaps. The authors consider that color Doppler ultrasound is one of the most useful diagnostic tools. Three cases are reviewed in which this technique aided the decision whether re-exploration was necessary. PMID- 11886999 TI - Distraction osteogenesis in Pierre Robin sequence and related respiratory problems in children. AB - INTRODUCTION: Sleep apnea is one of the most frequent manifestations of respiratory obstruction. Historically this clinical entity has stimulated the production of numerous valuable contributions with one purpose in mind: the improvement of airway permeability. A multi-disciplinary approach is required to define the problem in anatomic and functional terms to avoid a tracheostomy and prevent long-term sequels. We decided on an approach that focuses on improving the projection of the tongue in the posterior pharynx; by lengthening the mandible and bringing the muscular insertions of the floor of the mouth forward, the antero-posterior dimensions of the airway are increased. OBJECTIVE: To evaluate mandibular distraction osteogenesis as a simple mandibular lengthening procedure useful as a definite treatment in patients with obstructive sleep apnea. MATERIAL AND METHODS: The series consisted of 15 consecutive patients, divided in four groups. Patients with acute upper airway obstruction who required endotracheal intubation, patients with no acute upper airway obstruction but with severe respiratory distress, patients with milder degrees of airway obstruction, and patients with long-term tracheostomies. Therapeutic interventions were performed according to the findings of each group. RESULTS: The patients were evaluated according to cephalometric analysis, polysomnograms, nasopharyngoscopy, and clinical data. Significant changes were seen in the cephalometric studies. There were no postoperative episodes of apnea; clinical improvement occurred in all patients and decannulation was possible in all patients. CONCLUSION: Mandibular distraction is a safe and reliable procedure for treating patients with obstructive sleep apnea. PMID- 11887001 TI - CD44 as prognostic factor in oral and oropharyngeal squamous cell carcinoma. AB - This retrospective case control study was conducted to assess the prognostic value of some patient-, tumor-, treatment-related variables, and to correlate markers of primary tumor with survival and cervical metastases. Twenty-five patients with histologically proven squamous cell carcinoma of the oral cavity and oropharynx were analyzed. Patients were never treated before and had a minimum follow-up review of 45 months. Results show that T-stage is the most important clinical prognostic parameter. Regarding immunohistochemical markers (Ki67 and CD44), only CD44 seems to be significantly correlated with prognosis but this value showed a multicollinear effect with N upon survival. Decreased expression of CD44 correlates with a decreased survival, although increased CD44 expression was consistent with a longer survival. Therefore, it was assessed that a loss of cell adhesion, related to decreased expression of CD44, may be determinant of survival in these patients. PMID- 11887002 TI - Fully endoscopic vascular decompression of the glossopharyngeal nerve. AB - Microvascular decompression of the glossopharyngeal nerve is an effective treatment of patients with glossopharyngeal neuralgia in whom compression of the nerve by a blood vessel is implicated in the pathogenesis of the disease. The standard surgical technique uses a binocular operating microscope for intra operative visualization. Growing experience with posterior fossa endoscopy, however, has suggested that endoscopes may provide more comprehensive anatomical views of cerebellopontine angle. This report describes the case of a patient suffering from glossopharyngeal neuralgia who underwent fully endoscopic vascular decompression of the glossopharyngeal nerve. During this procedure the endoscope was used to survey the posterior fossa, guide the placement of insulating sponges, and conduct a final assessment of the intervention. We found the endoscope ideally suited to the constricted operating space of the posterior fossa, allowing for accurate localization and careful separation of the pathological vascular conflict with minimal brain retraction and no damage to surrounding structures. The versatility of endoscopy allows for superior visual appreciation of neurovascular conflicts in the posterior fossa. To date, endoscopy has primarily been used to supplement microscopy in cranial nerve decompression surgery. This report demonstrates how the endoscope can be used as the sole imaging modality in glossopharyngeal nerve decompression, with excellent results. PMID- 11887003 TI - Mandibula distraction osteogenesis for lengthening the mandibula to correct a malocclusion: a more than 70-year-old German concept in craniofacial surgery. AB - Bone distraction is still on the rise again since McCarthy et al. presented in their clinical investigation new osseous formation in the elongated area while performing the distraction of the mandible in 1992. But at the level of craniofacial skeleton, the initial description to the technique of distraction osteogenesis should be credited to the German craniofacial surgeons Rosenthal for the bone lengthening of the mandible in a microgen patient in 1927. The procedure is described and the original schedules and case are presented. PMID- 11887004 TI - Long-term neuropsychological effects of sagittal craniosynostosis on child development. AB - The link between cranial deformity and "functional" disability is not obvious in single-suture sagittal craniosynostosis. Physicians have anecdotally reported that children with simple craniosynostosis often seem to have a higher proportion of learning disabilities and cognitive problems than their nonafflicted peers. These problems have not been systematically studied, however. This study examined the long-term neuropsychological effects of single-suture sagittal craniosynostosis on selected aspects of neurological development. It did so by going beyond global measures of mental function (intelligence quotient) in an attempt to assess the incidence of subtle neuropsychological sequelae. Retrospective inspection of the Yale Department of Neurosurgery records between 1980 and 1990 was used to identify study subjects born with nonsyndromic sagittal suture craniosynostosis who were between 6 and 16 years of age at the time of the study. Of the 16 study subjects born with sagittal synostosis, which is thought to be among the most benign of the single-suture craniosynostoses, this study found that 50% had a reading and/or spelling learning disability. Although children with single-suture sagittal craniosynostosis fall within the normal range for intelligence, there is a significantly higher incidence of learning disabilities in this group than in the general population. PMID- 11887005 TI - Differences between direct (anthropometric) and indirect (cephalometric) measurements of the skull. AB - This study sought to determine the relative reliability of indirect cephalometric measurements and direct anthropometric ones taken in norma frontalis of 25 dry adult human skulls. Six of the 11 linear projective measurements were singular and located in the orbital, middle, and lower parts of the face, with two from each part. Two of the five paired measurements were taken in the orbital region on both sides, and the other three were taken in the middle to lower face between the midpoint of the facial axis and the landmarks lateral to it. Both singular and paired cephalometric distances were significantly shorter than the anthropometric distances. Mean numerical differences were much greater in paired measurements than in singular ones. The differences between these two sets of findings are a result of the uneven position of the landmarks used for measurement, located, as they are, on different planes of the face. These differences are undetectable by two-dimensional cephalograms. PMID- 11887009 TI - The IXth International Congress, International Society of Craniofacial Surgery, Visby, Gotland, Sweden, June 2001. PMID- 11887007 TI - Closure of critical sized defects with allogenic and alloplastic bone substitutes. AB - PURPOSE: This study evaluates bone regeneration of critical sized cranial vault defects in New Zealand white rabbits using four commercially available bone substitutes: OsteoSet (calcium sulphate pellets), DynaGraft Putty (demineralized bone matrix delivered in a poloxmer excipient), Norian CRS, and Bone Source (two commercially available calcium phosphate cements). MATERIALS AND METHODS: Critical sized defects 15 mm in diameter were created bilaterally in the parietal bones of 30 adult male New Zealand White rabbits. They were divided into three groups with ten animals in each. Bone healing was assessed clinically, radiographically, and histomorphometrically. Group 1 had calcium sulfate bioimplant on one side of the calvarium and an unfilled defect on the contralateral side. Group 2 had DBM putty on one side and Poloxamer gel on the contralateral side. Group 3, the Calcium phosphate cements (CPC), had Norian CRS on one side and Bone Source on the contralateral side. Five animals in each group were killed at 6 weeks and 12 weeks post operatively. RESULTS: All unfilled defects healed with fibrous scar, as did the Plaster of Paris and the poloxamer gel defects. Defects reconstructed with the demineralized bone matrix putty healed with bone throughout the entire defect. This was obvious clinically and radiographically where the defects appeared completely filled with a dense radiopaque tissue. The six-week group displayed new bone formation (87.1%) surrounding the remaining allogeneic particles. Resorption was evidenced by the presence of osteoclastic activity and by the significant decrease in the size of the demineralized bone particles. By 12 weeks, the demineralized bone putty bioimplant was almost completely replaced by new bone (95.5%). Both calcium phosphate cement groups (Norian CRS and Bone Source) had identical patterns of healing. They clinically were visible and firm and uniformly radiopaque with little evidence of new bone formation. Histologically the cement remained unresorbed with little new bone with in the defect at 12 weeks. CONCLUSIONS: The utilization of a demineralized bone matrix putty appeared to allow for complete closure of critical sized calvarial defects in New Zealand white rabbits with viable new bone at 12 weeks. PMID- 11887012 TI - Development of the facial midline. AB - "Intellectual excellence lies in having faith in the observation of apparently nontranscendental and unimportant facts. To observe an anatomic element calmly, with an open, analytical spirit, and with spiritual freedom, can lead to an explosive vortex of new knowledge."-Miguel Orticochea, M.D.(1) Traditional descriptive embryology based upon the interaction of frontonasal, lateral nasal, and medial nasal prominences is incapable of explaining the three-dimensional development of the facial midline. The internal structure of the nose and that of the oronasal midline can best be explained by the presence of paired A fields originating from the prechordal mesendoderm, associated with the nasal and optic placodes, supplied by the internal carotid artery, and sharing a common genetic coding with the prosomeres of the forebrain. Mesial drift of these fields leads to fusion of their medial walls; this in turn provides bilateral functional matrics within which form the orbits ethmoids, lacrimals, turbinates, premaxillae, vomerine bones, and the cartilages of the nose. This two-part paper reports six lines of evidence supporting the field theory model of facial development: (1) An apparent watershed exists in the midline of the base between the territories of the internal and external carotid systems. Isolation of the ICA in injected fetal specimens confirmed that the demarcation was distinct and restricted to the embryonic nasal capsule. (2) Field theory explains the developmental anatomy of the contents of the nasal capsule. (3) The neuromeric model of CNS development provides a genetic basis for the anatomy and behavior of fields. (4) Mutants for the Dlx5 gene demonstrate A field deletion patterns. These experiments relate the nasal placode to the structures of the A fields. (5) Separate regions of the original nasal placodes give rise to neurons, which are dedicated to separate sensory and endocrine systems. The A fields constitute the pathways by which these neurons reach the brain. (6) Non-cleft lip-related cleft palate, holoprosencephaly, and the Kallmann syndrome are clinical models that demonstrate the effects of anatomic disturbances within the A fields. PMID- 11887014 TI - HPV in "carcinoma in situ" associated with HIV/AIDS infection: a case report. AB - Annual incidence of condyloma acuminata has been reported to be high as 106.5 per 100.000, or about 0,1% of the entire population. In HIV+ individuals, HPV infection is even more prevalent 20% in HIV+ homosexual men and 27% in AIDS homosexual men). HPVDNA 16 and 18 are the most commonly found in anogenital condyloma and in 1% of pre-malignant lesions and oral cancer. OBJECTIVE: The aim of this publication is to report a clinic case of an AIDS patient with spreading condylomas in oral mucosa, one of these diagnosed as "carcinoma in situ". CASE REPORT: A 46 year-old male diagnosed with AIDS due to a history of intestinal Mycobacterium avium, esophageal candidiasis and Citomegalovirus (CMV) retinitis. He came to our consultation as an out-patient with a healthy appearance under highly effective antiretroviral therapy (HAART). On the exploration we found spreading condylomas in all of the oral mucosa. Biopsies were performed and histological and immunohistochemical studies revealed the characteristic features of HPV infection. PCRRLFP assay was added in order to obtain a sensible typification of HPVDNA. One localization of condyloma, in the upper lip mucosa was diagnosed as "carcinoma in situ". PVDNA types 18 and 51 were confirmed. Therapy was establish with Laser carbon-dioxide application in three sessions, followed by surgical excision with electro scalpel. Carcinoma in situ required excision surgery. A local therapy was performed and follow up. CONCLUSION: We have not find any report about HPVDNA 51 in oral "carcinoma in situ". Early detection of HPV lesions in oral mucosa is important in improving prevention of oral cancer. PMID- 11887015 TI - Odontogenic cysts. Analysis of 856 cases. AB - Odontogenic cysts (OC) are one of the main causes of jaw destruction. Information about these lesions in the Mexican population is scant. And for this reason the purpose of this work is to describe the frequency of the different varieties of OC recorded in two oral pathology services in Mexico City. As well as to compare the findings with those previously reported in other studies and to analyze the association of these lesions with the gender of the affected patients and the type of oral pathology service. There were a total of 856 OC; of these, 449 (52.5%) occurred in men, 403 in women (47%), and in 4 cases (0.5%) gender was not stated. There were 8 out of the 10 different types of OC recognized by the WHO. The most frequently diagnosed OC were radicular cyst (342 cases), dentigerous cyst (283 cases) and odontogenic keratocyst (184 cases). Together, these three entities represented 94.5% of all OC. Both the gender and the type of oral pathology service showed a significant association with radicular and dentigerous cysts (p<0.01). The knowledge of the origin, clinico-pathological features and the biological behavior of these lesions are basic aspects to achieve an early diagnosis and a proper treatment. PMID- 11887016 TI - Clinical evaluation of hard tissue proliferations in the mouth. AB - OBJECTIVES: This work shows the results of a study of the proliferation of bone tissue in the mouth. DESIGN OF THE STUDY: It was carried out on a sample of 530 persons who were natives of Central American countries, 179 males and 154 females, evaluating the rate of appearance of these formations depending on age, sex, racial group, and their location within the jaw. RESULTS: It was observed that 38% of the total sample showed hard tissue proliferation in the mouth. Of these cases, 158 (80%) were located in the superior maxilla and 48 (20%) of the cases in the jaw. Of which, 52 were men (25%) and 154 women (75%). This proliferation has greater incidence in the black race, in university students and liberal professions. The most frequent are torus palatinus and bilateral torus mandibularis. CONCLUSIONS: The Central American population shows a high incidence of hard tissue proliferations in the mouth; a radiological study is sufficient for their detection. PMID- 11887017 TI - Lesions of the oral mucosa in cocaine users who apply the drug topically. AB - The use and abuse of cocaine is increasingly frequent in many countries, and the associated problems are increasingly evident. The effects of cocaine in the oral cavity vary depending on the form used and the route of self-administration. In the present study we describe the lesions observed in four patients with a history of topical self-application of cocaine to the oral and/or nasal mucosa, with the aim of relieving pain produced by cocaine-induced cluster headache. In three of the four patients this practice has led to erythematous lesions, while the remaining patient showed gingival recession and bone sequestration. These lesions can probably be attributed to the vasoconstrictor activity of cocaine, and to its caustic effects on the mucosa. PMID- 11887018 TI - Clinical applications of the diagnosis of p53 alterations in squamous cell carcinoma of the head and neck. AB - Alteration of the p53 tumor suppressor gene implies an extremely high risk of developing malignancy, and mutation of the gene is one of the most frequent genetic changes found in human cancer. Squamous cell carcinoma of the head and neck (SCCHN) shows a high incidence of p53 tumor suppressor gene alterations; the latter therefore appears to play an important role in the pathogenesis and progression of such neoplasms. The loss of p53 protein activity may be due to many p53 gene mutations or to the action of certain viruses that infect the oral cavity. Local recurrence is the most common cause of mortality after SCCHN surgery; in this sense, p53 gene mutations have been observed in tissue adjacent to the tumor, and constitute a good prognostic marker of tumor recurrence. The analysis of p53 tumor suppressor gene alterations in SCCHN affords important information on the diagnosis, prognosis and treatment of affected patients - such alterations representing an indicator in high risk patients of the convenience of applying more aggressive adjuvant therapies. PMID- 11887019 TI - Prevalence of diabetes mellitus amongst oral lichen planus patients. Clinical and pathological characteristics. AB - OBJECTIVES: The aim of this study has been to define the prevalence of DM (considered according to the ADA-97 criteria) in OLP patients and also to investigate the existence of clinical and pathological differences between OLP patients with or without DM. STUDY DESIGN: Sixty-two patients suffering from OLP according to clinical and pathological criteria were selected to enter the study and classified after the DM diagnosis guidelines suggested by The American Diabetes Association (ADA-1997). The variables considered for each patient were: age, sex, clinical presentation, extension of the lesions, location of the lesions, number of locations, Candida albicans colonization, and density of subepithelial inflammatory infiltrate. RESULTS: Up to 27.4% of OLP cases were associated to type 2 DM (DM2) and 17.7% were related to an impaired fasting glucose (IFG). The mean age of the DM2-associated OLP group was significantly higher than the non-diabetic group's. No significant differences could be observed in terms of clinical and pathological features between diabetic and non diabetic OLP patients. CONCLUSION: The high prevalence of hydrocarbonate metabolism disorders observed in OLP patients justify the use of FPG for screening of DM2 in these patients. PMID- 11887020 TI - A comparative study between INR and the determination of prothrombin time with the Coaguchek(r) portable coagulometer in the dental treatment of anticoagulated patients. AB - OBJECTIVE: To assess the usefulness of the Coaguchek(r) portable coagulometer for determining the International Normalized Ratio (INR) in dental practice. STUDY DESIGN: A total of 139 INR determinations were made in 88 patients anticoagulated with acenocoumarol (Sintrom(r)) for thrombotic pathology, based on the habitual laboratory procedure (Sample 1). Posteriorly and prior to dental treatment, INR was again determined using the Coaguchek(r) portable device (Sample 2). Both determinations were subsequently compared to evaluate possible significant differences between them, applying the Student t-test for paired data and regression measures. RESULTS: The mean INR in Sample 1 was not significantly different to that recorded with the Coaguchek(r) portable device (Sample 2) (2,31 0,81 versus 2,28 0,82, respectively, t= 0,98; p= 0,32). A statistically significant relation was observed between the two samples (R= 0,92; p< 0,01). CONCLUSIONS: The Coaguchek(r) portable coagulometer is a valid instrument for determining INR in anticoagulated individuals, and constitutes an effective method in application to the outpatient dental treatment of such patients. PMID- 11887022 TI - A retrospective study (1994-1999) of 441 ITI(r) implants in 114 patients followed up during an average of 2.3 years. AB - OBJECTIVE: Implantation is a common practice in the oral rehabilitation of edentulous patients, affording good results in extensive patient series. The aim was to determine the average success rate and analyze the causes of implant failure. MATERIAL AND METHODS: In the present retrospective study (1994-1999) a total of 441 ITI(r) implants were performed in 114 patients (58 females (51%) and 56 males (49%) aged 44.5 years on average; range 12-82 years), with a mean follow up of 2.3 1.3 years. Data recorded included patient age and sex, smoking habit, oral hygiene, the number, length, diameter and type of implants, their location and the prostheses fitted. RESULTS: After 2.3 1.3 years of follow-up, the implant success rate was 96.2%. In 7 patients a total of 15 implants failed during the osseointegration period, while in two patients a total of two implants failed following dentures placement. The failures were mainly attributed to over drilling during the surgical procedure, or to an overloading upon tightening in the osseointegration period. No significant relation was observed between implant loss and the rest of the clinical parameters studied, with the sole exception of implant length (failure increasing with shorter lengths)(chi-square; p = 0.001). PMID- 11887021 TI - Efficacy of two antiplaque and antigingivitis treatments in a group of young mentally retarded patients. AB - INTRODUCTION: Chemical management of dental plaque for controlling oral hygiene becomes necessary in high-risk patients such as the mentally retarded. MATERIAL AND METHODS: Thirty-seven mentally handicapped patients aged 10-19 years and with severe plaque and gingivitis were divided into two treatment groups: Group I (daily mouthrinse with triclosan-zinc for 8 weeks) and Group II (0.2% chlorhexidine spray for 2 weeks). Both groups were evaluated at the start of the study and after 2 and 8 weeks. RESULTS: Significant reductions in plaque were observed in Group I after two weeks, with very significant improvements in both plaque and gingivitis after 8 weeks. In Group II, highly significant reductions in both indices were recorded after two weeks of treatment - significance persisting after 8 weeks. CONCLUSIONS: Triclosan-zinc mouthrinse and chlorhexidine spray can be effective adjuncts to tooth brushing for controlling dental plaque and gingivitis in mentally retarded patients. PMID- 11887023 TI - Characterization of Clostridium perfringens strains isolated from Polish patients with suspected antibiotic-associated diarrhea. AB - BACKGROUND: The aim of our research was to investigate the role of enterotoxin- producing anaerobic bacteria other than Clostridium difficile in the etiology of antibiotic-associated diarrhea. This article presents data related to C. perfringens. MATERIAL/METHODS: Stool samples taken from 158 patients with suspected antibiotic-associated diarrhea were specifically cultured for Clostridium difficile, Bacteroides fragilis and Clostridium perfringens. In order to associate the presence of virulence factors in the bacterial isolates thus collected with disease features, all strains were genetically and phenotypically analyzed for toxin production. All isolated C. perfringens strains were cultured in Ellner sporulation-promoting medium. RESULTS: In 21 of the 158 patients (13%) C. perfringens could be cultivated from the fecal specimen. None of the strains produced enterotoxin, and consequently the cpe gene was not detected by PCR in any of these strains. C. perfringens and C. difficile were cultivated from the same stool samples in 4 cases. Interestingly, in one case toxin A-negative/toxin B positive C. difficile and non-enterotoxigenic C. perfringens were co-cultured. After application of a heat shock (100 degrees C at 30 min.) only two C. perfringens strains producing thermoresistant spores were detected. Pulsed field gel electrophoresis (PFGE) demonstrated genetic heterogenicity among the C. perfringens strains, suggesting that these bacteria were already presented upon hospital admission. CONCLUSIONS: It seems unlikely that nosocomial transfer has taken place. The relatively low incidence suggests that C. perfringens is not a major primary cause of antibiotic-associated diarrhea. PMID- 11887024 TI - Effect of oral supplementation of free amino acids in type 2 diabetic patients-- a pilot clinical trial. AB - BACKGROUND: Oral amino acid intake reduces plasma glucose in Streptozotocin induced diabetic rats. This study examined the effect of oral amino acid supplementation in patients with type 2 diabetes mellitus (DM). MATERIAL/METHODS: A double blind pilot clinical trial was conducted for a period of 2 months on 77 subjects with type 2 DM. Subjects of both sexes, ages 30-60, were included in the trial. All were receiving oral antidiabetic tablets. They were divided into groups on the basis of oral supplementation: (A) lysine, (B) essential amino acids, (C) amino acids and vitamins (fat and water-soluble), and (D) calcium phosphate (control). The subjects were periodically examined for fasting and post prandial plasma glucose, fasting and post-prandial immunoreactive insulin, plasma amino acids, glycosylated haemoglobin (HbA1c), proteins and albumin in serum, urea and creatinine in plasma and sugar, and proteins and ketones in urine. RESULTS: The results revealed a significant decrease in PP plasma glucose (P<0.05) in group B when compared to groups C and D after 45 days. Plasma Arginine was increased in group C from 3.84 to 9.24 mg/dl. There were no statistically significant changes seen in other parameters between groups and visits. CONCLUSIONS: Oral supplementation with amino acids for patients with type 2 DM appears to decrease PP plasma glucose without any change in plasma insulin levels, perhaps due to improved insulin sensitivity. However, the long term effects of amino acids need further study. PMID- 11887025 TI - Clinical implications of acute myocardial infarction complicated by high grade atrioventricular block. AB - BACKGROUND: The purpose of this study was to assess the incidence, clinical course, prognosis and mean length of stay in acute myocardial infarction (AMI) complicated by high-grade atrioventricular block (HAVB). MATERIAL/METHODS: A retrospective cohort study including all AMI patients listed from January 1995 to September 2000 in the ARIAM multi-center register. Univariate analysis was carried out to study the factors associated with the development of HAVB, the mortality rate, and the mean length of stay, and multivariate logistic regression analysis to study whether HAVB is an independent predictive variable for mortality or prolongation of stay. RESULTS: Of the 14,181 AMI patients included in the register, 837 (5.9%) presented with HAVB, which was associated with age, female sex, increased severity, diabetes, inferior and Q-wave AMIs, and a higher peak creatine phosphokinase (CPK) level. The HAVB patients developed more complications, required more diagnostic-therapeutic resources, and showed significantly higher mortality (p<0.0001) and increased mean length of stay (p<0.0001). The independent risk factors for HAVB were age, maximum peak CPK, inferior or combined localization of the AMI, Q-wave AMI, diabetes, a Killip and Kimball score > 1, and thrombolysis. HAVB was found to be an independent predictive variable for mortality and increased mean length of stay. CONCLUSIONS: AMI patients with HAVB, despite thrombolytic treatment, are at risk for complications, mortality and longer mean admissions. Further study is needed on the outcome of a more active reperfusion policy, such as direct, rescue angioplasty etc. PMID- 11887026 TI - Circulating oxidized LDL is associated with increased levels of cell-adhesion molecules in clinically healthy 58-year old men (AIR study). AB - BACKGROUND: Inflammation has been postulated to play an important role in atherosclerosis development. Recently, research in this field has been focused on the role of modified lipoproteins, primarily oxidized low density lipoprotein. Adhesion and transendothelial migration of circulating leukocytes, by the action of cell-adhesion molecules, seem to be critical steps in early atherogenesis. Previous in vitro studies on the relationship between ox-LDL and the expression of cell-adhesion molecules on endothelial cells have shown diverging results. Furthermore, there are as yet no reports on the relationship between circulating ox-LDL and cell-adhesion molecules in vivo. MATERIAL/METHODS: The relationship between circulating ox-LDL and adhesion molecules was measured by ELISA in clinically healthy, 58-year-old men recruited from the general population (n=391). RESULTS: The results of the present study showed that circulating ox-LDL was associated with sICAM-1 and sE-selectin, but not with sVCAM-1. After adjustment for other risk factors, i.e. systolic blood pressure, LDL cholesterol, apoB, apoA1 and smoking, the observed relationships between ox-LDL and sICAM-1 and sE-selectin, respectively, remained statistically significant. CONCLUSIONS: The observed results fit into the concept that oxidatively modified LDL may regulate the increased expression of cell-adhesion molecules on endothelial cells in vivo. These observations were made in a population-based sample of clinically healthy, middle-aged Caucasian men. Since this was a cross-sectional study no conclusions can be drawn on causality. PMID- 11887027 TI - Long-term assessment of pulmonary function tests in pediatric survivors of acute respiratory distress syndrome. AB - BACKGROUND: The aim of the study was to evaluate long-term pulmonary function tests in pediatric survivors of acute respiratory distress syndrome (ARDS). MATERIAL/METHODS: Observational study based on a telephone poll of retrospectively identified post ARDS children who were hospitalized in a pediatric intensive care unit (PICU) in a general 1200-bed teaching, tertiary, regional referral center for children. RESULTS: Follow-up pulmonary function tests were achieved in only 7 children, with a mean age of 7.3+/-4.3 years (range 3-12) and following 5.6+/-4.3 years after PICU discharge. The etiology for ARDS included: lymphoma (n=2), pneumonia (n=2), aspiration (n=1), petrol ingestion (n=1) and snake envenomation (n=1). The children had been ventilated for 9.4+/ 7.3 days and their worst PaO2/FiO2 ratio was 65.1+/-17.0 mm Hg. The follow-up pulmonary functions in all the children was within normal limits except for one child who had mildly reduced DLCO and one who had mild exercise-induced hypoxemia (oxyhemoglobin saturation of 94%). Neither of the two nor the others showed subjective symptoms or clinical physical limitations. CONCLUSIONS: Children who survive ARDS apparently enjoy long-term normal pulmonary function. Some, however, may present subclinical dysfunction that persists for many years after the acute episode and evoked only by sophisticated lung tests. PMID- 11887028 TI - Inflammatory bowel disease in Greece-- a hospital-based clinical study of 172 consecutive patients. AB - BACKGROUND: It has been suggested that inflammatory bowel disease (IBD) has a milder course in Greece than in other Western countries. The purpose of our study was to assess the clinical picture of IBD in a hospital-based sample of IBD patients, followed from January 1986 to May 2001. MATERIAL/METHODS: Retrospective study of 172 patients: 130 with UC followed up for 1-360 months (mean 66+/-57 months) and 42 with CD for 1-240 months (mean: 51.85+/-63.68 months). RESULTS: 63 UC patients (48%) required hospitalization, 30 (23%) on the first attack; 72 (55%) had left sided colitis and 23 (18%) pancolitis; in 6 cases, (5%), surgical intervention was indicated at onset, 4 (3%) were operated. During FU, 15 patients (11.5%) were operated, 4 (3%) as medical emergencies, 3 (2%) due to colon cancer, the others for intractable colitis or long-standing disease; 5 patients (4%) developed toxic dilation of the colon, 3 (2%) developed colon cancer and 4 (4%) extraintestinal malignancies. 7 patients (5%) died; 2 (1.5%) due to the disease and the remainder for other causes. 19 CD patients (45%) had ileocolitis and 14 (33%) colitis; 22 (52%) needed hospitalization on the first attack. In 12 patients (29%) surgical intervention was required on initial onset or during FU. One patient developed colon cancer (2%). Two men died (5%) from myocardial infarction. CONCLUSIONS: IBD does not have a milder course in Greece than in other European populations. PMID- 11887029 TI - When is total aortic arch replacement indicated in patients with acute aortic dissection? AB - BACKGROUND: The purpose of this preliminary study was to evaluate the effectiveness of our surgical strategy for acute aortic dissection with special emphasis on curative resection of the dissected segments of the aorta. MATERIAL/METHODS: Between January 1995 and April 1999, 29 patients underwent surgery for acute aortic dissection. In 16 patients (Group 1) the dissection was limited to the ascending aorta (8 patients - Group 1a) or involved the ascending aorta and the entire aortic arch (8 patients - Group 1b). Complete resection of all dissected aortic segments (ascending aorta or ascending aorta with complete aortic arch) was performed in these cases, extending to the healthy tissue border. 13 patients (Group 2) presented with dissection of the entire aorta). These patients underwent replacement of the proximal part only of the dissected aorta. RESULTS: Early mortality (within 30 days) and the incidence of perioperative cerebrovascular events was 3.4% and 10.3% respectively. These events all occurred in Group 2. During the follow-up period of up to six years, there were no significant differences between the surviving patients in regards to long-term mortality and morbidity, although a persisting patent false lumen was observed in seven patients from Group 2. CONCLUSIONS: Extension of ascending aorta replacement to include the complete aortic arch can be accomplished in patients amenable to complete resection of the dissected aorta without increasing operative risk and with good mid-term results. We believe that total aortic arch replacement is indicated in these cases. PMID- 11887030 TI - Assessment of the value of pHmetry results in diagnostics of gastroesphageal reflux as a cause of obstructive bronchitis in children. AB - BACKGROUND: The study assessed the incidence of pathologic gastroesophageal reflux in children with obstructive bronchitis and analyzed the correlation of pHmetric parameters with the incidence of obstructive bronchitis according to age groups. MATERIAL/METHODS: 109 children aged from 4 months to 17 years with the history of at least two documented incidents of obstructive bronchitis during the period of one year were examined. Each patient underwent 24h esophageal pHmetry. The analyzed patients were divided into 3 age groups: I - 4 months-2 years, II - 3-9 years, III - 10-17 years; each of the obtained groups was divided into 2 subgroups according to the frequency of obstructive bronchitis (a - 2-4 times a year and b - over 4 times a year). RESULTS: Gastroesophageal reflux was diagnosed on the basis of pHmetry in 62.4% of patients. It was demonstrated in own material that gastroesophageal reflux is more frequent in boys (64.7%) than in girls (35.3%). Comparison of pHmetric parameters in children suffering from obstructive bronchitis more and less frequently did not demonstrate higher pHmetric parameters in children with more frequent occurrence of obstructive bronchitis in any of the age groups. Although gastroesophageal reflux occurred frequently in children with obstructive bronchitis, no proportional correlation was found between the pHmetric parameters and frequency of obstructive bronchitis CONCLUSIONS: Esophageal pHmetry should be performed in children with obstructive bronchitis in order to confirm or exclude gastroesophageal reflux as a cause of recurrent obstructive bronchitis. PMID- 11887031 TI - Role of fasting serum C-peptide as a predictor of cardiovascular risk associated with the metabolic X-syndrome. AB - BACKGROUND: Insulin resistance with increased insulin and C-peptide levels is the basis of the metabolic X-syndrome, so it is reasonable to expect them to be a good predictor of associated cardiovascular risk factors. MATERIAL/METHODS: A total of 29 patients (21 postmenopausal women and 8 men) with type 2 diabetes mellitus (mean duration 14.6 years, 95% CI 11.9 to 17.3 years), all older than 50, were studied for possible links between fasting serum C-peptide levels and other vascular risk factors. The mean value of the C-peptide in the group was 0.627 nmol/l (95% CI: 0.464 to 0.789 nmol/l). RESULTS: We found statistically significant correlations between C-peptide and triacylglycerols (TG; r=0.474; p=0.009), HDL-cholesterol (inverse; r = -0.567; p = 0.001) and various lipoprotein ratios: atherogenic index (= total/HDL cholesterol: r = 0.599; p = 0.0006) or TG/HDL (r = 0.587; p = 0.0008). C-peptide also correlated with the body mass index (BMI: r = 0.519; p= 0.004) and leptin (r = 0.492; p = 0.007). After the coefficient CpG (C-peptide x fasting glycemia) was introduced, the correlations with lipoproteins became even stronger. CONCLUSIONS: We suggest that elevated (fasting) serum C-peptide levels constitute a clinically important marker of the cardiovascular risks associated with the metabolic X-syndrome. It can be used as an effective tool for the early detection of diabetic patients at particular risk for atherosclerotic cardiovascular diseases and needing early preventive measures or aggressive treatment. PMID- 11887032 TI - Further studies of genetic susceptibility to Graves' disease in a Russian population. AB - BACKGROUND: Graves' disease (GD) is a polygenic autoimmune thyroid syndrome. Some of the genes implicated in its pathogenesis may encode thyroid-stimulating hormone receptor (TSHR) and estrogen receptors 1 (ESR1) and 2 (ESR2). We examined dinucleotide repeat polymorphisms in the ESR1 and ESR2 genes and D727E amino acid substitution in the TSHR gene for possible association with GD in a Russian population. MATERIAL/METHODS: The polymorphic regions of the target genes were amplified by polymerase chain reaction (PCR) on the basis of genomic DNA isolated from blood of 78 unrelated Russian patients with GD and 93 control subjects. To detect the D727E TSHR polymorphism, the PCR product was additionally digested with Eco72I restriction endonuclease. The genotype and allele frequencies in the groups studied were compared by c2 test. The odds ratios and 95% confidence intervals (CI) were calculated to assess the strength of the relationship between the polymorphisms tested and GD. RESULTS: For polymorphic dinucleotide microsatellites at ESR1 and ESR2, no significant difference was observed in allele frequencies between affected and nonaffected patients. For the D727E TSHR polymorphism, the E allele and the DE genotype were significantly more frequent (p<0.0001) in patients with GD than in control subjects. CONCLUSIONS: The D727E variant of the TSHR gene is associated with Graves' disease in a Russian population. The E727 allele and the heterozygous D727E genotype are related to higher risk of the disease. No association with GD was found for polymorphic microsatellites of the ESR1 and ESR2 gene. PMID- 11887033 TI - Assessment of correlation between the presence of antiendomysial antibodies and small intestine mucosal villous atrophy in the diagnostics of celiac disease. AB - BACKGROUND: The aim of the study was to analyze the degree of correlation between the presence of serum antiendomysial antibodies and villous atrophy in small intestine mucosa in patients with celiac disease. MATERIAL/METHODS: Retrospective analysis carried out in a group of 88 children and young adults (28 males, 60 females aged from 2 to 23) hospitalized in the Clinic from 1990 to the present with positive results of serum antiendomysial antibody test performed because of suspected celiac disease. The IgA - EmA test was performed after determination of whole IgA level in the serum using indirect immunofluorescence technique with monkey esophagus as an antigen. All patients underwent endoscopic small intestine biopsy with histopathological assessment of mucosal bioptates using standard hematoxylin and eosin staining. RESULTS: The serum levels of IgA in all investigated patients were normal. The levels of antiendomysial antibodies expressed in IF units ranged from 10 to 40960. Histopathological assessment of small intestine mucosal bioptates revealed partial (in 13.6% of patients) or total (in 86.4% of patients) villous atrophy. CONCLUSIONS: The study demonstrated complete correlation between the presence of serum antiendomysial antibodies and villous atrophy in the small intestine mucosa. PMID- 11887034 TI - Selected parameters of fibrinolysis system in patients with dermatitis herpetiformis. AB - BACKGROUND: Dermatitis herpetiformis (DH) is a very rare bullous dermatitis of autoimmune origin. It is a syndrome involving dermal and intestinal pathology, in which vesicopapular skin lesions are accompanied by gluten-dependent enteropathy. The diagnosis of DH is based on immunopathological investigation of unaffected skin bioptate (presence of granular IgA deposits in the upper portions of dermal papillae). MATERIAL/METHODS: The study was carried out in a group of 33 patients, including 23 males and 10 females aged 22-78 (mean age 44.7 years). The following fibrinolysis system parameters were determined in the sera of all patients: tissue plasminogen activator concentration (t-PA: Ag), urokinase plasminogen activator concentration (u-PA: Ag), plasminogen activator inhibitor concentration (PAI-Ag), plasminogen level, alpha-2 antiplasmin activity (alpha-2-AP), plasmin-- alpha-2 antiplasmin complexes concentration (PAP). RESULTS: Mean t-PA concentration in DH patients was 5.52 ng/ml, vs. 4.8 ng/ml in controls. The respective u-PA concentrations amounted to 0.33 ng/ml, and 0.39 ng/ml. PAI-1 concentration was markedly higher in DH patients (36.2 ng/ml) than in controls (22.40 ng/ml), whereas plasminogen level was significantly lower (86.0% vs. 115.9%). In patients, alpha-2-AP activity was 92% and was lower than in controls 103.4%. DH patients demonstrated also higher concentrations of PAP complexes (327.45 ng/ml) than the control group (203.03 ng/ml). CONCLUSIONS: Patients with DH were found to have lower plasminogen and alpha-2 antiplasmin levels and increased concentrations of PAP complexes, which indicates increased plasminogenesis in vivo. Increased type 1 plasminogen activator inhibitor level (PAI-1) may reflect a chronic inflammatory condition present in DH patients. PMID- 11887035 TI - Credibility of problem-solving therapy and medication for the treatment of depression among primary care patients. AB - BACKGROUND: Patient beliefs in the effectiveness of treatment may have an important influence on treatment outcome. MATERIAL/METHODS: Associations between patient beliefs in the credibility of treatment and outcome were explored in a randomised controlled trial of major depression in primary care (n=155). The four treatments were antidepressant medication given by research general practitioner, problem solving treatment given by research general practitioner or research practice nurse over 12 weeks or a combination of problem solving treatment and antidepressant medication. Patients' belief in the credibility of treatment was assessed using a brief Credibility Scale, that was completed following randomisation and after treatment. Depression outcome was measured at 6, 12 and 52 weeks using the Hamilton Rating Scale for depression, and the Beck depression inventory. RESULTS: Pre-treatment, medication treatment was associated with a higher certainty of recovery than was problem-solving treatment from the nurse (p=0.018). Post-treatment, medication and combination treatment were seen as more logical than problem-solving treatment from the nurse (p<0.03). Post-treatment medication had higher certainty of recovery and was more highly recommended to a friend. Linear regression demonstrated that the depression outcome measures were not associated with either pre- or post-treatment credibility. CONCLUSIONS: Patients found all four treatments highly credible following their initial explanation. There was a significant difference both pre- and post-treatment in favour of patients finding treatment involving medication more credible than problem-solving from a nurse. Pre- and post-treatment scores of credibility were not associated with outcome. PMID- 11887036 TI - Comparison of three instruments for assessing ongoing intimate partner violence. AB - BACKGROUND: Active injury surveillance programs need to address 'ongoing' intimate partner violence (IPV). While the Abuse Assessment Screen (AAS) has been validated for 'present'(within a year) IPV it is not clear that it is valid for 'present' (ongoing) IPV. We have created an OAS (Ongoing Abuse Screen, OAS) by changing the AAS to specifically request information related to 'ongoing' IPV. The hypothesis of this study was that the OAS represents a construct that is different from either the original AAS or a single question asking about ongoing IPV. MATERIAL/METHODS: All patients presenting to the ED during a convenience sampling of shifts completed the survey. The survey included the OAS, the AAS, and the question 'Are you presently a victim of IPV?' Comparisons were made between these 3 using the kappa statistic for agreement. RESULTS: A total of 488 surveys were completed. The AAS was positive in 288/488 (59%, 95%CI= 55-63%), the OAS was positive in 78/488 (16%, 95%CI=13-19%), and the single question for DV was positive in 14/488 (3%, 95%CI=2-5%). Kappa was 0.28 for the AAS and the OAS. When compared to the single question about present DV, kappa was 0.05 for the AAS and 0.27 for the OAS. CONCLUSIONS: The OAS may be a useful tool for evaluating ongoing IPV. The OAS resulted in rates different from that of the AAS and may be more specific to ongoing IPV than the AAS and more sensitive than a single question about DV. PMID- 11887037 TI - Effect of erythropoietin therapy and selenium supplementation on selected antioxidant parameters in blood of uremic patients on long-term hemodialysis. AB - BACKGROUND: The kidney accumulates the highest level of selenium (Se) in the organism and is the major source of plasma glutathione peroxidase (GSH-Px). Se, as an integral part of the active site of GSH-Px, plays an important role in protecting cell membranes from oxidative damage. Decreased blood Se levels and GSH-Px activity are common in chronic renal failure (CRF) patients. Our study was an effort to evaluate the effect of erythropoietin (EPO) therapy and Se supplementation for CRF patients undergoing regular hemodialysis (HD) on blood Se, red cell glutathione (GSH), and blood lipid peroxidation product levels, and on blood activity levels of GSH-Px and blood superoxide dismutase (SOD). MATERIAL/METHODS: Our subjects were divided into three groups: I - CRF patients on regular HD and EPO, II - HD patients receiving EPO and Se, and III - healthy controls. Se levels, SOD and GSH-Px activities were measured spectrofluorometrically, the GSH level by Beutler's colorimetric method, and lipid peroxidation products using TBARS. RESULTS: EPO therapy with Se supplementation significantly increased whole blood and plasma Se in HD patients, and raised red cell GSH-Px activity, but plasma GSH-Px activity, plasma superoxide dismutase, and plasma and red cell TBARS did not respond to Se supplementation. EPO alone showed no effect on these parameters. CONCLUSIONS: Treatment with EPO and supplementation with Se significantly increased the element concentration in whole blood and plasma, and GSH-Px activity in red cells. Plasma GSH-Px activity did not respond to Se. PMID- 11887038 TI - Radioguided parathyroidectomy for reexploration of primary hyperparathyroidism-- a case report. AB - BACKGROUND: We report a case of radioguided parathyroidectomy using a hand-held gamma probe for the reexploration of primary hyperparathyroidism. CASE REPORT: The patient was a 66-year-old Japanese woman. She had previously undergone surgical exploration for primary hyperparathyroidism due to a left inferior parathyroid tumor detected by 99mTc-methoxyisobutylisonitrile (MIBI) scintigraphy. However, the pathological diagnosis of the resected tumor was adenomatous goiter. 99mTc-MIBI scintigraphy was performed again and revealed an abnormal uptake close to the right lower lobe of the thyroid. However, venous sampling for PTH measurements did not support this finding. Sestamibi was injected and the radioactivity was measured pre- and intraoperatively with a hand held gamma probe. With the patient under general anesthesia, the tumor, which was adjacent to the right recurrent laryngeal nerve, was resected, but it contained only a low level of radioactivity ex vivo, indicating that it was not a parathyroid tumor. A hand-held gamma probe accurately located the radioactive parathyroid tumor in the right lower neck. The resected tumor measured 15 x 6 mm and weighed 331 mg. The pathological diagnosis was parathyroid adenoma. CONCLUSIONS: Radioguided parathyroidectomy is useful to localize parathyroid tumors not only in primary hyperparathyroidism at the initial neck exploration but also for reexploration. PMID- 11887039 TI - Cat-scratch disease in an immunocompromised host. AB - BACKGROUND: The main causative agents of cat-scratch disease are Bartonella henselae, tiny, gram-negative bacilli. The disease usually has a benign course with the development of a papule at the inoculation site, followed by regional lymphadenopathy. In most cases, complete resolution occurs, but in immunocompromised hosts, the course of the disease can be aggravated. CASE REPORT: A patient received methotrexate and corticosteroids for 3 months due to rheumatoid arthritis. He developed fever, exanthema and leukopenia under methotrexate therapy. Dark red indurations with central ulcerations at his right thigh revealed a further problem apart from the methotrexate-induced leucopenia and immunosuppression. The ulcerations were the remainders of recurrent scratches from the patient's cat. The patient's antibody titers against Bartonella henselae remained low and inguinal lymph node swelling was only for a short time to be observed, this reaction obviously weakened as a result of the immunosuppression. However, the typical course, the exclusion of other reasons for the exanthema and the rapid improvement of the patient's condition after antibiotic treatment ascertained the diagnosis. CONCLUSIONS: In immunocompromised hosts, diseases with a typically benign course can become severe and life-threatening illnesses. Ownership of pets should be taken into consideration before onset of an immunosuppressive therapy. PMID- 11887040 TI - Designing the optimal Total Cavopulmonary Connection: pulsatile versus steady flow experiments. AB - BACKGROUND: The Total Cavopulmonary Connection (TCPC), used for repair of patients with single ventricle physiology, creates a passive system of blood flow into the pulmonary circulation where enhanced energy efficiency may lead to improved long term patient outcomes. Previous numerical and in vitro studies using steady flow have shown that incorporation of SVC (superior vena cava) and IVC (inferior vena cava) offsets lead to decreased energy losses. We hypothesize that the optimal TCPC offset design found in these previous steady flow experiments may not be the optimal design in pulsatile flow situations. MATERIAL/METHODS: 3-D finite volume numerical models were used to simulate flow through the total cavopulmonary connection. We ran steady and pulsatile flow experiments through 4 TCPC designs each with different SVC to IVC offsets (0.1/4, 1/2, 1 diameter offsets). The total energy (power) loss for each TCPC model was calculated. RESULTS: In steady flow experiments, % difference in energy loss between the most optimal and least optimal design was 26%. In contrast, in pulsatile flow experiments the % difference was only 8%. CONCLUSIONS: Our results demonstrate the improvements in energy loss seen using SVC-IVC offsets in steady flow experiments do not necessarily translate to pulsatile flow situations. Overall there was lower differences in efficiency between all TCPC designs in the pulsatile flow experiments. These results emphasize the need for further studies to fully define the relationship between energy losses and TCPC vessel architecture in non-steady flow physiologic situations. PMID- 11887041 TI - Fractal dimensions of human brain cortex vessels during the fetal period. AB - BACKGROUND: Fractal dimensions always involve some averaging with respect to the given geometrical object. The classical geometrical description includes overly detailed geometrical information, rendering the precise formulation of equations ineffective. The problems associated with the functionality and growth of biological systems according to their observed geometrical shape require a specific approach. MATERIAL/METHODS: The material consists of 50 brains, 4th to 8th month of fetal life, length C.-R. 83-220 mm. The material was examined using the Pickworth method, computer image analysis (Imtronic, Scion for Windows 1998, and Elf v. 4.2), and fractal analysis. Various transformations were applied: numeric filters, statistical filters, binary geometrical transformations, arithmetical operations. Fractal analysis employs the 'box-counting' fractal dimension. RESULTS: The cortex vein net maintains the fetal character, with a large number of anastomoses, also present in adults. The size of cortex branches amounts to 100-200 microm. The reduction of some of the anastomoses during the early stages of evolution comes from an increase in the vessels' caliber. During the 4th month, the fractal dimension is 1.26, increasing to 1.53 at month 5. Rapid growth of the fractal dimension has been observed until the 6th and 7th months. This means an increased complexity level and feed volume in the brain. CONCLUSIONS: Fractals constitute a promising and effective tool to estimate the structures of brain vessels. The fractal dimension varies during the fetal period. This shows the close correspondence between the structure and functionality. PMID- 11887042 TI - Liver regeneration: the emergence of new pathways. AB - The present overview summarizes important knowledge having accumulated during the last years. The liver maintains a steady mass which is basically controlled by a delicate balance between cell gain and cell loss. However, reconstitution of the organ after tissue loss does not only involve replacement of target cells, but also complex remodeling processes resulting in the reconstruction of the typical tissue architecture. Most information in liver regeneration refers to hepatocytes. It is important to note that hepatocytes are not terminally differentiated cells, but cells situated in the G0 phase that can undergo proliferation upon appropriate stimulation. In most situations, hepatic stem cells are not significantly involved in this response. Hepatocyte regeneration is accomplished by a sequence of distinct phases: an initiation phase, rendering cells in a state of replicative competence; a proliferation phase, where expansion of the cell population occurs; and a termination phase, where cell growth is suppressed to terminate regeneration at a set point. These three phases are regulated by a whole group of factors, mainly cytokines, the significance of which has in part been defined by use of animal models with target gene deletions (gene knockouts). It seems that several mechanisms are capable to sense the critical cell mass which has to be achieved. Hepatocyte regeneration is accompanied by a complex remodeling of hepatic tissue, with a transient breakdown of the lobular architecture. In contrast to hepatocytes, less is known for the the regenerative replacement of bile ducts, blood vessels and hepatic stellate cells. PMID- 11887043 TI - Asthma and gastroesophageal reflux in children. AB - Gastroesophageal reflux (GER) is a factor often neglected in the etiopathogenesis of asthma. The estimated incidence of GER in asthmatic children reaches 50-60% and is higher than in the general population. GER may accompany typical symptoms: hoarseness, sore throat, thoracic pain, cough or wheezing. GER may not only aggravate the course of bronchial obstruction, but may also cause it, or trigger obstruction due to other factors. Asthma and GER coincidence has been acknowledged for many years. The paper presents a current review of studies concerning the relations between asthma and GER and attempts to establish, which is the cause and which is the result. The hypotheses how GER can lead to bronchial obstruction, and how obstruction can aggravate GER, are also presented. GER is believed to be a factor causing obstruction by: 1. an indirect mechanism - reflex theory, 2. a direct mechanism - reflux theory, and 3. a neuropeptide mediated mechanism. The paper also presents diagnostic methods allowing to detect GER in asthmatics. A review of recent studies concerning the treatment of GER in asthmatics, both with pharmacological and surgical methods, is also included. Beneficial effect of antireflux therapy on the course of asthma has been emphasized. Therefore, antireflux therapy is recommended in all patients with concurrent asthma and GER, irrespective of severity of clinical GER symptoms, even in those with silent GER. The essential drugs used in the treatment of GER are proton pump inhibitors. Appropriately high dose level and appropriately long duration of the therapy should be taken into consideration. PMID- 11887044 TI - The artificial heart-- past, present, and future. AB - Patients with advanced congestive heart failure have a very high 5-year mortality despite medical treatment. In such patients, heart transplantation is the treatment of choice. The number of patients awaiting transplantation is several fold higher than the number of procedures performed. Heart transplantation therapy has numerous limitations and is associated with serious complications. The left ventricular assist device is a step towards this goal. It can be attached to the weakened left ventricle to temporarily increase blood flow to the body. The use of left ventricular assist devices helps the failing heart to recover and extends the duration over which a patient's heart can wait for a replacement donor heart. This therapy is in use for only up to a few months. The total artificial heart, Jarvik-7, first implanted in 1982, did not succeed due to a poor quality of a patient's life and numerous complications leading to death. Recently, a successful implantation of the AbioCor (Abiomed), the first fully implantable replacement heart, was accomplished. The AbioCor's internal battery system eliminates the need for the patient to be permanently immobilized through tubes or wires connected to an external power source. Innovative transcutaneous energy transmission permits the recharging of internal batteries. The total artificial heart will require adapting it to different human body sizes as well as further improving its technical features. The total artificial heart is a remedy of the future coming to fruition right now, giving a chance to numerous heart failure patients by extending and improving their lives. PMID- 11887045 TI - Use of MIDCAB procedure for redo coronary artery bypass. AB - BACKGROUND: Reoperative coronary artery bypass grafting (CABG) procedures are growing in importance due to the increasing number of patients requiring another bypass operation. Conventional redo-procedures are associated with an increased mortality and morbidity. To reduce risk, minimally invasive direct coronary artery bypass (MIDCAB) using the left internal mammary artery (LIMA) to the left anterior descending branch (LAD) may be preferable, when indicated, in selected patients. We report a series of patients who underwent this procedure for redo CABG in our center. METHODS: Since April 1997, 20 male patients who had undergone prior CABG using conventional procedure, were reoperated using the LIMA to LAD through a lateral minithoracotomy on the beating heart. Nineteen patients presented for a redo-CABG; one patient required a second-time redo-CABG. Two patients required concomitant PTCA of a second vessel as hybrid procedure. We reviewed these redo cases and studied their surgical results for mortality, morbidity, operation time, and hospital stay. RESULTS: Mean operation time was 139 min (90-180). Four patients were extubated directly postoperatively; the others had a short period of ventilatory support. There was no myocardial infarction, no deaths or need of inotropic support postoperatively. No patient required re-exploration for bleeding. All patients could be mobilized and discharged early. At present, all patients are living and classified as CCS class I or II. CONCLUSIONS: Our results indicate that MIDCAB using IMA grafts for reoperation is a safe procedure with low risk for morbidity and mortality. This surgical technique is a useful alternative to conventional redo CABG in selected patients when complete revascularisation is not indicated. PMID- 11887046 TI - Mitral valve replacement in ischemic mitral regurgitation. Preservation of both anterior and posterior mitral leaflets. AB - BACKGROUND: The surgical risks associated with ischemic mitral regurgitation are thought to be greater than those for other forms of mitral regurgitation. We have performed mitral valve replacement using the St. Jude Medical bileaflet prostheses with preservation of both leaflets, along with all of the chordae tendineae and papillary muscles. The aim of this study was to retrospectively evaluate mitral valve replacement with preservation of both mitral valves with respect to long-term clinical results and left ventricular performance. METHODS: Between January 1, 1988 and February 29, 2000, 15 patients were operated on for ischemic mitral regurgitation. There were 7 males and 8 females, and the mean age was 69.7+/-8.1 years. The preoperative variables showed clinical deterioration of the state, such as emergency operation in 40% of the patients, more than NYHA functional III class in 93% of patients, cardiogenic shock in 47% of the patients, a mean left ventricular ejection fraction of 36.8%, and a mean left ventricular end-systolic volume index of 116.7 ml/m2. RESULTS: There were 5 (33.3%) hospital deaths during the follow-up period including 1 early death and 1 (10%) late death during the follow-up period. Thus, the actuarial survival rate after 5 years for the whole was 60%. However, the left ventricular dimensions and left ventricular fractional shortening, even if in patients with profound depressed left ventricular function preoperatively, showed maintenance of the cardiac function. CONCLUSIONS: These results suggested that mitral valve replacement using the St. Jude Medical prostheses with preservation of both leaflets and all chordae tendineae and papillary muscles might be a procedure of choice for ischemic mitral regurgitation. PMID- 11887047 TI - Troponin I release after CABG surgery using two different strategies of myocardial protection and systemic perfusion. AB - BACKGROUND: Controversies still exist over the optimal temperature for blood cardioplegia and systemic perfusion. This study investigates the effect of temperature of blood cardioplegia and systemic perfusion on the release of troponin I and other biochemical markers. METHODS: One hundred and fifty-four consecutive patients were randomly assigned to one of two cardioplegic and systemic perfusion strategies of cold blood cardioplegia with moderate systemic hypothermia (27 degrees C) or tepid blood cardioplegia with mild systemic hypothermia (33 degrees C). Cardiac troponin I and other biochemical markers were measured at baseline, at the end of surgery, at 12 hours and daily thereafter. A two-way ANCOVA for repeated measure was performed to test the effect of cardioplegia on enzyme release independently of variables that were different between the two groups. RESULTS: The time course of dismission of troponin I, creatine kinase MB, and lactate dehydrogenase were significantly lower with tepid blood cardioplegia and mild systemic perfusion independently of the number of distal anastomoses, CPB time, cross clamp time or total volume of cardioplegia. There were no differences between the two groups in the release of total creatine kinase, aspartate transaminase and alanine transferase. CONCLUSIONS: Both strategies of myocardial protection and systemic perfusion guarantee subclinical minor myocardial damage. The strategy of tepid whole blood cardioplegia and mild systemic hypothermia seems to preserve myocardium better than whole blood cold cardioplegia. PMID- 11887048 TI - Concentrated platelets harvesting before cardiopulmonary bypass improved cardiac and pulmonary function. AB - BACKGROUND: Sequestration of concentrated platelets (P-con) during cardiopulmonary bypass (CPB) has been performed to preserve platelet function after cardiac surgery. Since P-con also harvests leukocytes simultaneously, there might be a possibility that the inflammatory effects or ischemia-reperfusion injuries associated with CPB, such as a cardiac or pulmonary dysfunction after cardiac surgery, are reduced with its use. METHODS: We retrospectively evaluated 53 patients who underwent cardiac surgery after the introduction of the P-con technique at our institute. There were 20 patients in the P-con group and 33 patients in the control group in whom concentrated platelet were not harvested. RESULTS: The patients characteristics and preoperative cardiac and pulmonary function did not differ between the two groups. The percentages of platelets and leukocytes sequestrated were 20.2+/-5.4% and 8.5+/-3.9% of the total estimated circulating cell counts, respectively. There were no significant differences in the postoperative dose of dopamine used, cardiac index, pulmonary capillary wedge pressure or intubation period between the two groups. However, the stroke volume index (p=0.005), left ventricular stroke work index (p=0.002), and ratio of the arterial oxygen tension to the inspired fraction of oxygen on extubation (p=0.02) were significantly greater in the P-con group as compared with those in the control. CONCLUSIONS: P-con improved cardiac and pulmonary function after CPB. Simultaneous sequestration of platelets and leukocytes by P-con during CPB may contribute to the improvement of cardiac and pulmonary function after cardiac surgery. PMID- 11887049 TI - Correlation between age and vital organ function following deep hypothermic circulatory arrest. AB - BACKGROUND: We investigated the correlation bet-ween perioperative variables such as patients' age and vital organ function after operation on thoracic aorta using deep hypothermic circulatory arrest (DHCA). METHODS: Ninety-five patients who underwent replacement of thoracic aorta under DHCA for acute or chronic aortic dissection and aortic aneurysm, and survived more than 10 days were divided into group I (age less than 60, n=17), group II (between 60 and 69, n=39), and group III (over 70, n=39). Concomitant procedures such as aortic root replacement and coronary artery bypass grafting were performed in 9, 4, and 1 patients in group I, II, and III, respectively. Postoperative pulmonary, renal, and hepatic function within 10 days were compared. Correlation between other perioperative variables and organ function was also investigated among all 95 patients. RESULTS: Postoperative pulmonary, renal, and hepatic function in group III was not significantly inferior to those in groups I and II. The operation time, and amount of red blood cells and fresh frozen plasma used during operation, were significantly greater in group I, which showed an intimate correlation to significant elevation of hepatic enzymes. CONCLUSIONS: Although lower functional reserve of vital organs in the elderly patients was predicted, they showed an acceptable functional recovery after operation with DHCA. Other perioperative variables such as operation time and blood transfusion showed a negative impact on postoperative hepatic function. PMID- 11887051 TI - Coronary artery bypass grafts in situs inversus. AB - Coronary artery bypass grafting in dextrocardia is rare. A case is described for the first time where both arterial and venous conduits were used. The subtle changes required in the conduct of the operation are highlighted. PMID- 11887050 TI - Intravenous allicin improves pulmonary blood flow after ischemia-reperfusion injury in rats. AB - BACKGROUND: Allicin is a sulfur-containing compound extracted from garlic, with antiaggregatory, anti- migratory, anti-oxidant and pulmonary vasodilator actions. We hypothesized that allicin might be beneficial in lung ischemia-reperfusion. METHODS: A non-nothermic rat lung ischemia-reperfusion model was established by clamping left pulmonary artery (PA) for 1 hr, followed by reperfusion for 2 hrs by clamping right PA to reflect solely the function of left lung. Groups were control (n=7), allicin 0.1 mg (n=8) and allicin 0.01 mg (n=4). In the beginning of reperfusion allicin/saline were injected. Pulmonary artery pressures (PAP), pulmonary artery flow (PAF), left atrial pressure (LAP) were monitored. At the end of reperfusion period arterial blood gas (ABG) analysis was done. RESULTS: Six of 7 control and 3 of 8 group 2 animals died before completing the experiment. In group 1 all animals completed the experiment (p=0.015 vs control). PAF was significantly increased after 30, 60 and 120 min of reperfusion in group 1 (p=0.0028, 0.0009, 0.0003 respectively vs control) and after 60 and 120 minutes in group 2 (p=0.0453, 0.018 respectively vs control). Pulmonary vascular resistance was lower at 30 min in allicin 0.01 mg group (p=0.0017 vs control). PAP was increased after 60 and 120 min of reperfusion in group 1 (p=0.016, 0.0029 respectively vs control) and after 120 min in group 2 (p=0.0104 vs control). CONCLUSIONS: This study shows that allicin improves postischemic PAF in this model. Allicin needs further investigation of potential utility and mechanism(s) of action. PMID- 11887052 TI - Washing of the residual solution of cardiopulmonary bypass circuit after coronary artery bypass grafting in idiopathic thrombocytopenic purpura. AB - A 76-year-old female with chronic idiopathic thrombocytopenic purpura required coronary bypass grafting. Preoperative treatment with high-dose intravenous immunoglobulin at a dose of 0.4 g/kg/day raised the platelet count from 57,000 to 110,000/microL. After termination of cardiopulmonary bypass (CPB) the residual blood in the CPB circuit was washed to reduce total immunoglobulin G (IgG) level, including platelet-associated immunoglobulin G (PA-IgG), and returned to the patient. Intraoperative platelet transfusion was used due to a drop in platelet count to the pretreatment level. The postoperative level of IgG and PA-IgG remained significantly lower than preoperatively. The postoperative course was uneventful and without bleeding complications. Perioperative management of a patient with idiopathic thrombocytopenic purpura undergoing open-heart surgery is discussed. PMID- 11887053 TI - Emergency management of spontaneous coronary artery dissection. AB - Six cases of spontaneous coronary arteries dissection are reported. In one patient, triple vessel spontaneous coronary artery dissection was identified. Another patient presented spontaneous left main coronary artery dissection. In one case we found the spontaneous dissection of the left anterior descending artery associated with distal aortic arch dissection. These conditions are very rare and may present a surgical dilemma. Causative factors and underlying pathology are clarified. Prompt diagnosis and surgical intervention is safe and effective. Early recognition of left main coronary artery dissection or three vessel dissection is essential because urgent coronary artery bypass grafting may be life saving. PMID- 11887054 TI - Metastatic renal cell carcinoma to the right ventricle complicating thrombocytopenia. AB - Solitary right ventricular metastasis of renal cell carcinoma is extremely rare. Herein, we report a surgical case of metastatic renal cell carcinoma to the right ventricle, associated with an unusual presentation of "a syndrome resembling idiopathic thrombocytopenic purpura". PMID- 11887055 TI - Supramitral ring: an unusual cause of congenital mitral stenosis. Case series and review. AB - Supramitral ring, also known as membranous supravalvular mitral stenosis is a rare cause of congenital mitral stenosis, with less than 100 cases appearing in the literature since its first description in 1902. We present a small series encountered at the university medical center during the last five years. The natural history of the condition is reviewed along with diagnostic tools, aspects of surgical repair, and anesthetic technique that facilitates early extubation and ICU discharge. PMID- 11887056 TI - Left ventricular pseudoaneurysm after acute influenza A myocardiopericarditis. AB - We describe the successful management of a rare case regarding an adult Caucasian who developed a left inferior ventricular pseudoaneurysm as a consequence of an influenza A virus infection of the upper respiratory tract followed by acute myocardiopericarditis. The cardiovascular features of this case illustrate the importance of raising a sufficient clinical index of suspicion for this common, but potentially lethal, entity. PMID- 11887057 TI - Abdominal and thoracic aortic aneurysms. How should we treat them? PMID- 11887058 TI - Management of aortic graft infection. AB - BACKGROUND: The optimal method of operative treatment of prosthetic aortic graft infection (PAGI) has been the subject of debate; incidence rates of PAGI are low. Diagnosis of PAGI can be difficult. The aim of this retrospective study is to evaluate our results in treating PAGI in order to try and optimize the treatment of this grave problem. METHODS: Thirty-eight patients (median age 68.5 years) were treated for PAGI between 1991 and 2000. Management of PAGI was performed with total graft excision and simultaneous extra-anatomic bypass (n=18), total graft excision and in situ repair with a Rifampicin-soaked gelatin-impregnated prosthetic aortic graft (n=8), or a partial excision with in situ repair (n=11). In 1 patient, only local irrigation was performed. The median follow-up was 45 months. RESULTS: Clinical presentation of PAGI (median interval 3 years) was: discomfort/pain (n=14), gastro-intestinal bleeding (n=11), persisting fever (n=8), or a non-healing wound (n=5). The primary patency rate in patients with extra-anatomic bypass was 67% at 6 months follow-up. In patients with other surgical reconstructions no graft occlusion was encountered. Overall amputation rate was 5%. Recurrent infection of the graft was 15%. The overall early mortality rate in this study was 21%. CONCLUSIONS: The diagnosis of PAGI is difficult and should be based on a combination of clinical symptoms, laboratory findings and imaging techniques. There are several treatment options that should be tailored to the extent of infection and the patients' physical condition. In a selected group of patients partial excision of the infected graft only can be justified. PMID- 11887059 TI - Lymphoceles complicating arterial reconstructions of the lower limbs: outpatient conservative management. AB - BACKGROUND: Treatment of lymphoceles complicating vascular procedures is controversial. The purpose of this study was to evaluate the practicability, effectiveness and safety of conservative management on an outpatient basis. METHODS: Medical records were reviewed for 23 postoperative lymphoceles (in 18 patients) seen at our institution between 1986 and 1999. Diagnosis was made by physical examination and needle aspiration of fluid collection; bacterial cultures were obtained in all. Ultrasonography was performed in all patients, lymphoscintigraphy (99mTc HSA) in 11, angio-TC in 2 cases, MRI in 3 large lymphoceles. RESULTS: Twenty-one lymphoceles developed in the groin, 2 in the thigh and were mostly (72.2%) diagnosed after hospital discharge. Imaging techniques detected subcutaneous wound collection; in addition, lymphoscintigraphy showed lymphatic interruption and collateral pathways in patients with limb swelling. Outpatient management consisted of limited ambulation, limb elevation and pressure dressings; no serial aspirations were made. Resolution was obtained in all patients over a mean period of 21 days (range, 12 to 35). No patient required re-hospitalization or developed wound and/or graft infection. No recurrence was noted after a follow-up of all patients for 1 year. CONCLUSIONS: Outpatient treatment of lymphoceles following arterial reconstructive procedures can be performed safely. Significant advantages of this pathway include no re-hospitalization and cost reduction. PMID- 11887060 TI - Spectral and profile analysis of Doppler recording following below-knee arterial distal bypasses. AB - BACKGROUND: Arterial below knee distal bypasses are associated with a high risk of thrombosis as compared to proximal bypasses. We assumed that before the bypass occludes, in the early postsurgical period, measurable velocity changes, and/or the presence of high intensity transient signals (HITS) would occur. METHODS: SETTINGS: institutional reference center, hospitalized patients. SUBJECTS: satisfactory Doppler recording was obtained in 51 among 61 consecutive patients (32 males, 19 females, height: 165+/-7 cm, weight: 68+/-12 kg) suffering lower extremity arterial disease, that underwent saphenous (n=33), prosthetic (n=4) or sequential (n=14) below knee bypasses. We performed a spectral and profile analysis of a single postsurgical 2 hour Doppler recording at the ankle level and analyzed Doppler derived indices and clinical risk factors in the evaluation of the risk of bypass occlusion within 7 days following surgery. RESULTS: Primary patency at day 7 was observed in 41 of the 51 operated patients. The presence of HITS was found in approximately 30% of the patients and provided no information on the risk of thrombosis. No clinical variable was significantly associated with an increased risk of thrombosis. Whatever the duration of recording, the presence of a diastolic forward flow and wide systolic velocity changes were poor indicators of bypass thrombosis risk. On 512 beat recordings, a mean systolic velocity below 1630 Hz and a standard deviation of the resistance index >0.095 were associated with a 6.74 [1.6-28.4] (p<0.01) and 14.5 [3.6-58.9] (p<0.001) times increases in the risk of bypass occlusion respectively, compared with subjects that do not fulfill each criteria. CONCLUSIONS: Periods of transient asymptomatic no-flow-reflow events may be observed before the bypass irreversibly occludes. Prolonged Doppler recording should be preferred to short term analyses, to allow for the detection of these transient events and may provide potential indices for future research. PMID- 11887061 TI - Popliteal venous aneurysms: involvement of tibial nerve in aneurysmal wall. AB - Primary popliteal venous aneurysms are very rare vascular abnormalities, with 50 cases reported in the English literature. Thromboembolic complications are common in popliteal venous aneurysms. Since medical treatment has been proved inadequate, surgical repair is recommended with a minimum time delay. However, it involves some risk because of the complex anatomy of the popliteal space. We report on a case of popliteal venous aneurysm, in which the tibial nerve was involved in the aneurysmal wall and was injured by surgical repair. PMID- 11887062 TI - Successful endovascular stent-grafting for thoracic aortic aneurysms in systemic lupus erithematosus. Report of 2 cases and review of the literature. AB - Systemic lupus erythematosus (SLE) is frequently associated with cardiovascular manifestations but rarely complicated with aortic disease, and surgical treatment is often complicated with later anastomotic dehiscence. We report successful endovascular stent-graft placement (EVSGP) as an alternative to conventional treatment of the aortic aneurysm in patients with SLE and review of the literature. Clinical cases included a 58-year-old woman with a saccular aneurysm of the distal aortic arch and a 52-year-old woman who had the aortic dissection in the whole descending thoracic aorta. Both patients underwent placement of the stent-graft in the diseased aorta through the iliac artery and received steroid perioperatively. Mortality was compared between surgical and medical treatment in the reported 39 cases of SLE associated with aortic aneurysm. Both patients were successfully treated by EVSGP and no inflammatory signs were seen after the procedure. There were no recurrence in the short follow-up period (up to 23 and 15 months after the procedure). In review of the literature, operative mortality (13.6%) was superior to that in patients receiving medical treatment only (53.3%), but two of 19 operative survivors died of rupture afterward. EVSGP can be a useful alternative to conventional treatment of the aortic aneurysm in patients with SLE, although it lacks the support of long-term follow-up data currently. PMID- 11887063 TI - The right retroperitoneal approach on abdominal aortic aneurysm with an isolated left-sided inferior vena cava. Report of a case. AB - We report herein the case of a 78-year-old man found to have abdominal aortic aneurysm (AAA) with an isolated left-sided inferior vena cava (IVC). The patient was admitted to our hospital to undergo surgery for the AAA. The computed tomography revealed the sacular aneurysm of the infrarenal abdominal aorta (60 x 40 mm) and right common iliac aneurysm (30 x 30 mm). At the same time the left sided IVC was found by the CT. This IVC (13 mm wide) ascended 76 mm, dorsally to the ureter, the left side of the AAA from the right common iliac artery to the left renal artery. We performed aneurysmectomy and 20 mm knitted Dacron bifurcating graft replacement by the right retroperitoneal approach without manipulating the left-sided IVC. The procedure was completed without incident and the patient has continued to do well. PMID- 11887065 TI - Detection of postoperative pseudoaneurysms following abdominal aortic aneurysm repair in Behcet's disease by MRA. AB - We report the development of two anastomotic pseudoaneurysms in a patient with Behcet's disease eighteen months after abdominal aortic aneurysm repair. Major asymptomatic vascular complications should be suspected in patients with Behcet's disease with a history of vascular surgery and treated expediently due to the risk of rupture. Magnetic resonance angiography, contrast-enhanced computed tomography or ultrasound scanning should be performed at least every 6 months after vascular surgery. PMID- 11887064 TI - Successful treatment of threatening limb loss ischemia of the upper limb caused by ergotamine. A case report and review of the literature. AB - We report a case of ischemia limited to the upper limb caused by chronic use of ergotamine. The effect of this drug was not related to hypersensitivity or to the potentiating effects of other medications. The patient was successfully treated discontinuing ergotamine and administering an alpha-blocking agent for three months. A review of the literature of the vascular complications of ergotamine is presented. PMID- 11887066 TI - Temporary spinal cord stimulation for peripheral cholesterol embolism. AB - Cholesterol embolism is often an unrecognized complication of some cardiac and vascular procedures (i.e. coronarography, angioplasty, aortocoronary bypass, abdominal aortic aneurysmectomy) and of therapies affecting coagulation (thrombolysis, anticoagulation). The degree of pain associated with ischaemic and necrotic lesions secondary to cholesterol embolism involving the lower limbs is disproportionate to the extension of tissue involvement. Spinal cord stimulation (SCS) has been recognized as effective in relief of pain of ischaemic and neuropathic nature, although its mechanism of action is still not completely clear. The authors are unaware of previous reports of peripheral cholesterol embolism treated by SCS. Two case reports of inferior limb ischaemia secondary to cholesterol embolism in patients who had undergone cardiac invasive procedures. Temporary surgical implantation of SCS devices, which were removed after 4 to 6 weeks. Pain relief was achieved within 1 to 4 hours of surgical procedure. Any analgesic medications could be immediately discontinued. Pain control was effective and normal daily activities were rapidly regained. Ischaemic lesions healed within 4 to 6 weeks of SCS. Pain control is the most critical aspect of the management of peripheral cholesterol embolism without visceral organ involvement. SCS provided effective pain relief in the reported cases and its established ability to improve peripheral microcirculation allowed rapid resolution of necrotic lesions. Temporary SCS should be considered in the management of painful necrotic skin lesions secondary to iatrogenic cholesterol embolism. PMID- 11887067 TI - Stapler blebectomy and pleural abrasion by video-assisted thoracoscopy for spontaneous pneumothorax. AB - BACKGROUND: This study aims to evaluate the efficacy of video-assisted thoracoscopic surgery (VATS) pleurodesis in the treatment of spontaneous pneumothorax with particular reference to the postoperative period and the rate of recurrence after pleural abrasion. METHODS: One hundred and thirty-three patients who underwent VATS management of primary spontaneous pneumothorax were retrospectively reviewed. They were 113 males and 20 females with median age of 26 (range 12-37). Among these patients, 114 underwent VATS for recurrent pneumothorax and 19 for persistent air-leakage at the first episode. During surgical procedure, in 78% of cases, parenchymal blebs were identified and resected by stapler resection. All patients were submitted to pleural abrasion. RESULTS: No intra- or postoperative deaths occurred. Postoperative complications were persistent air-leak for more than 7 days in 6 patients (4.3%) bleeding in 3 patients (2.2%). The median chest-tube duration and hospital stay were 2 (range 2 11) and 3 (range 3-12) days, respectively. Median follow-up period of 53 (range 6 96) months was complete for all patients. Five episodes of recurrent pneumothorax were encountered and 4 of them, because of major entity, required re-do VATS with stapler resection and pleural abrasion: their postoperative period and residual follow-up was uneventful. CONCLUSIONS: The goal in the surgical management of spontaneous pneumothorax, which often affects "apparently healthy" young patients, is to secure the less recurrence rate with no mortality and quite null morbidity and functional impairment. VATS stapler resection and pleural abrasion is a safe procedure allowing a good management of the disease with low complication rate, short chest-drain duration, hospital stay and recurrence rate quite similar to those referred for other procedures such as pleural poudrage or limited pleurectomy. PMID- 11887068 TI - Bronchopleural fistula after lung cancer surgery. Multivariate analysis of risk factors. AB - BACKGROUND: To elucidate retrospectively the risk factors for bronchopleural fistulae after lung cancer surgery. METHODS: The subjects were 1,177 patients with lung cancer who underwent surgery between 1983 and 1997. Twenty-two clinical factors were examined by logistic analysis. RESULTS: Bronchopleural fistulae were observed in 35 patients (32 males, 3 females) with a mean age of 64 years. Eighteen (51%) of 35 patients died of BPF-related complications. The significant risk factors obtained by univariate analysis were male gender, heavy smoking, current smoking, low level of %FVC, metastases to lymph nodes, squamous cell carcinoma, increased WBC, decreased albumin, advanced postsurgical stage, sleeve lobectomy, and resection of the right lower lobe or middle and lower lobe. The significant risk factors noted by multivariate analysis were heavy smoking (30 or more pack/years), current rather than past smoking, metastases to lymph nodes, decreased albumin (3.5 mg/dl or less), and resection of the right lower lobe or middle and lower lobe. CONCLUSIONS: The above risk factors must be taken into account before surgical techniques followed by adequate perioperative management are selected. PMID- 11887069 TI - Clinical and pathologic predictors of survival in patients with thymic tumors. AB - BACKGROUND: The aim of this study is to evaluate the impact of thymectomy in patients with thymic neoplasms and to identify clinical and histopathological factors associated with improved long-term outcome of surgery. METHODS: We treated 74 patients between February 1987 and July 1993. There were 29 total and 36 simple thymectomies. These last cases, all non-myasthenic, had benign thymomas (n=30) but 6 had thymic carcinomas. Nine tumors were no-resected (5 thymomas and 4 thymic carcinomas). Minimum follow-up by Department of Thoracic Surgery Istituto Nazionale Tumori was 60 months after thymectomy. We divided the specimens according to Marino and Muller-Hermelink's classification: 54 thymomas, 18 thymic carcinomas and 2 no-diagnosis specify thymomas. There were 53 stage I, 1 stage II, 13 stage III, 5 stage IVa and 2 stage IVb according to Masaoka. RESULTS: Forty-six patients with treated thymoma were alive without disease at the end of follow-up, the remaining 8 died from recurrence in 6, a new tumor in 1 and a heart attack in the last. Of 18 thymic carcinomas 9 were alive at the end of follow-up (1 with recurrence), only 4 dead from recurrence. The actuarial survival of patients with thymomas was 88.5% at 5 years, (73.6% in cortical type, 85.7% in medullary type, 93.9% in mixed type, 100% in predominantly cortical type). Myasthenia gravis didn't influence the survival: 87.3 (no MG) vs 90%. Advanced stage thymomas significantly increased the risk of death from early stage I: 32.4 vs 100% at 5 years. In thymic carcinoma patients with well differentiated thymic carcinoma (WDTC) died less than others: the actuarial probability of survival at 5 years was 90 vs 68%. CONCLUSIONS: Thymectomy was the best treatment to long term outcome. In our experience, survival was related to histotype and to local extension of tumor. PMID- 11887070 TI - Tissue engineered tracheal prosthesis with acceleratedly cultured homologous chondrocytes as an alternative of tracheal reconstruction. AB - BACKGROUND: Autologous tissue is an ideal substitue for an extensive tracheal reconstruction, but it is rarely feasible in clinical situations. Many tracheal prosthesis had been used for such an instances, but unfortunately it is still problematic. Dislocation, local infection, hemorrage, and luminal stenosis can cause prosthetic failure. To achieve clinically available autologous tracheal prosthesis, it is necessary that we have to get phenotypically functioning chondrocytes, rapid differentiation of harvested autologous chondrocytes, and the survival of free grafted cultured chondrocytes. METHODS: In this study, we investigated isolation and culture method of the chondrocytes using the rabbit costal cartilage, and the cells were characterized microscopically and biochemically first. Then we have used cultured rabbit chondrocytes to investigate the role of growth factors upon the proliferation and regulation of the cultured chondrocytes. We have examined the effect of peptide growth factors on DNA and proteoglycan synthesis to the rabbit chondrocyte. The effects of IGF-I and basic FGF were investigated individually. Secondly, acceleratedly cultured chondrocytes were embeded to polymer (PLGA) scaffold in bioreactor, and implanted to defected rabbit trachea. Six weeks later, the rabbits were sacrificed and examined their histologic characteristics. RESULTS: The harvested chondrocytes from costal arch grew well and were amplified successfully maitaining their own phenotypes. Its embedding to PLGA scaffold was accomplished successfully. The implanted tracheal prosthesis maintains its physical integrity well, but the histologic examination revealed non-viable chondrocytes. The epithelial linings were good. CONCLUSIONS: The tissue engineered tracheal prosthesis can be a promising alternative of good functional air way tube in short term experiment, but biologically not vital yet. Further investigations are necessary to see the survival of free grafted chondrocytes and the long term results. PMID- 11887071 TI - Prosthetic reconstruction of the trachea in rabbit. AB - BACKGROUND: Various tracheal reconstruction techniques have been developed for stenosis. Different types of grafts, flaps and synthetic materials have been used for reconstruction of the defect when primary anastomosis was unsuccessful or not indicated. The mentioned reconstruction methods have limited success. Polytetrafluoro-ethylene (PTFE) prosthesis is a microporous polymer and has been applied for implantation on a wide range. It is also appropriate for tracheal reconstruction. METHODS: In the present study, segmental defects were created in 12 New Zealand rabbits. The rabbits were divided into 2 subgroups; the first group was applied an end-to-end anastomosis whilst the second a PTFE prosthesis. After 2 months, these applications were compared clinically, endoscopically and histopathologically to each other. RESULTS: Necrosis and extrusion were not observed in the rabbits with PTFE applications. After 1 month, the tracheal stenosis was found on endoscopic examination in 5 animals in the first group and 2 animals in the second group. While in longer defects, end-to-end anastomosis causes tracheal tension, PTFE applications have been well tolerated. CONCLUSIONS: It is concluded that PTFE prosthesis is a suitable alternative method in reconstruction of circumferential tracheal defects. PMID- 11887072 TI - Severe obstruction of the superior vena cava caused by tumor invasion. Recanalization using a PTFE-covered Z stent. AB - Bare stents are commonly used for the treatment of malignant vena cava stenoses. However, the therapeutic effect of treatment using bare stents for cases with intraluminal tumor invasion is not satisfactory. We report a case with severe obstruction of the superior vena cava caused by tumor invasion of mediastinal lymph node metastases from colon cancer, which was successfully treated by the recanalization of superior vena cava using a polytetrafluoroethylene-covered Z stent. The covered stent could not be fully expanded at first, and re-obstruction developed at the stented site due to thrombus formation soon after stenting. So, the additional balloon dilatation made the stent expend fully on another day of stenting. After the balloon dilatation blood flow improved immediately and the clinical symptoms associated with the superior vena cava obstruction resolved. Thereafter no symptomatic recurrence has been observed in 12 months of follow-up period. PMID- 11887073 TI - Invasive pulmonary aspergillosis with intraspinal extension in a non immunocompromised host. A case report. AB - A case of invasive pulmonary aspergillosis in a 33-year-old, non immunocompromised male presenting with lower limb paraplegia is reported. The clinical manifestations, diagnosis, and combined medical and surgical management of this rare condition are described. PMID- 11887075 TI - Prognostic trend in advanced implant surgery. AB - BACKGROUND: Implant surgery in association with bone grafting is generally considered less predictive than primary implant surgery. Many reports have been published about implant rehabilitation with bone grafts in atrophic patients. Most of these papers showed a lower implant success rate than primary implantology. The aim of this study is to verify if it's possible to warrant similar results between the two types of implantology, if such procedures are performed following effective protocols and criteria. METHODS: From 1995 to 1999, 43 severely atrophic edentulous patients were treated in our Clinic with 63 autologous bone grafts and delayed implantology; 45 patients were treated with traditional implantology. 284 fixtures were positioned. The success rate in grafted implantology versus traditional implantology was compared for every maxillary and mandibular region. Furthermore, success rate in implantology of the anterior maxilla versus the poster maxilla in grafted patients was compared. The statistical considerations were performed with c2 test (p<0.05). RESULTS: The statistical analysis evidenced not significative difference in the implant success rate between grafted and not grafted patients in the anterior (p=0.23) and in the posterior maxilla (p=0.35). There was not significative difference in the implant success rate between grafted and not grafted patients in the anterior mandible (p=0.54) and in the posterior mandible (p=0.54). There was not significative difference in the implant success rate between the anterior and posterior grafted maxilla (p=0.21). CONCLUSIONS: The results obtained show that if close surgical protocol is performed it is possible to obtain no prognostic difference between the two METHODS. PMID- 11887074 TI - Pericardial flap aortopexy: an easy and safe technique in the treatment of tracheomalacia. AB - A 5-month-old boy who had been operated for esophageal atresia and tracheoesophageal fistula was presented with recurrent life-threatening apneic spells, expiratory stridor and difficulty in feeding. Diagnosis of tracheomalacia was confirmed by bronchoscopy and pericardial flap aortopexy was performed. Pericardial flap aortopexy is a relatively simple procedure with minimal risk to the aorta. Minimal dissection is required and there are no sutures placed in the aortic wall, thus avoiding the risk of tears. PMID- 11887076 TI - [Statistical survey of afferent pathologies during a 5-year study in the oral pathology Department at the Second University of Naples]. AB - BACKGROUND: Oral pathologies represent an increasingly frequent clinical entity and are often the local manifestations of systemic pathologies. The purpose of this study was to carry out a statistical and epidemiological survey of the afferent pathologies in the department of oral pathology at the Second University of Naples from December 1995 to December 2000. METHODS: A total of 2112 patients were studied, divided by pathology and on the basis of valid epidemiological criteria. A statistical survey was carried out using different tests based on the nature of data available. RESULTS: This survey showed a marked predominance of lichen cases (347), but it is important not to underestimate the significant incidence of BMS (burning mouth syndrome) (284) which exceeded the frequency of aphthous lesions (258) and leukoplakial lesions (218). CONCLUSIONS: Mouth diseases account for a significant part of the disorders found in clinical dentistry. This study focused on those pathologies with the highest frequency, basing this analysis on etiological and clinical criteria and stimulating the interest of the dentist in individual forms by highlighting their incidence during normal professional activities. PMID- 11887077 TI - [Periodontal disease and occlusal trauma: a still debated controversy? A review of the literature]. AB - In the "Glossary of Periodontics Terms" written by the American Academy of Periodontology, the occlusal trauma is defined as "an injury to the attachment apparatus as a result of excessive occlusal forces". Nowadays, the effects of occlusal trauma on tooth support tissues, the onset and the progression of periodontal disease are still debated: many commonplaces have been disproved, but some doubts and not yet clear points remain, even owing to the difficult diagnosis of the presence and the real clinical impact of a traumatic occlusion. Then, ethical reasons prevent researchers from prospective clinical trials. At the beginning of the last century occlusal trauma has been supposed to be an etiologic factor of "alveolar pyorrhea", but several studies attending more strict scientific criteria failed to prove such correlation. On the basis of the bacterial genesis of periodontal disease, researchers started evaluating the possible effects of occlusal discrepancies on incidence, progression and treatment outcomes of periodontitis, but all the results underlined the more relevant role played by micro-organisms. The present review of the literature runs through this controversy again, analysing the most significant studies published. PMID- 11887078 TI - [The use of osteodistraction in treatment of ankylosis of the temporomandibular joint. A case report]. AB - The authors evaluate the possible use of osteodistraction techniques to restore the vertical dimension of the mandibular ramus after reduction during surgical treatment of TMJ ankylosis. They report the case of a fully grown female patient with monolateral right TMJ ankylosis following previous fracture injury who was treated at the Maxillofacial Surgery Unit of George Eastman Hospital in Rome. The patient underwent removal of the ankylotic block and the concomitant restoration of the vertical dimension of the mandibular ramus achieved using subperiosteal osteodistraction. Bichat's bubble flap was used as the interarticular interposition material. The distractor was activated one week after surgery with daily increments of 1 mm for a total of 7 days. Clinical and instrumental tests performed after the removal of the distractor and after a cycle of physiotherapy confirmed the good functional recovery of the joint and the restored vertical dimension of the mandibular ramus. The authors affirm that the method used represents a valid alternative for correcting loss of height after extensive resection of ankylotic joint processes in fully-grown patients. PMID- 11887079 TI - [Neuromuscular occlusion in the orthodontic treatment of craniomandibular disorders. A clinical case]. AB - In the last few years, it has been observed a considerable increase of cranio mandibular disorders which presently represent one of the most common commitments for dentists and, in particular, for orthodontists. The aim of this paper is to evaluate the efficacy of Jankelson's therapeutic protocol, followed by an orthodontic treatment, in a patient with cranio-mandibular disorders associated with occlusal pathologies. The cranio-mandibular electromyography and kinesiography promoted by Jankelson provide objective diagnostic measurements, while tens, relaxing masticatory muscles, allows a correct recording of myocentric occlusion. In this case, the authors utilise these devices, according to a well established diagnostic and therapeutic protocol. The first step is purely gnatologic, and consists of the application of an orthotic to temporarily treat pain and dysfunction. The following phase is a simple orthodontic treatment representing the final therapy. Orthodontic and/or prosthodontic rehabilitation of dentition, in fact, is the ultimate step of the therapeutic scheme which allows long-lasting RESULTS. PMID- 11887080 TI - [Nodular fasciitis in the zygomatic area. Case report and review of the literature]. AB - The authors report a clinical case of nodular fasciitis in the zygomatic area. Although this tumor-like proliferation occurs very rarely in the maxillofacial region, careful diagnosis based on histologic examination must distinguish it from neoplastic lesions such as fibrosarcoma with which the disorder shares certain histologic features. The case is compared with others described in the literature. In June 1996 a 12-year-old girl presented at our unit with swelling of the zygomatic region and positive history for trauma to the area. Palpation of the soft tissues detected a nodular formation about 1.5 cm in diameter. Echography and magnetic resonance imaging confirmed the presence of a nodular proliferation above the bone surface. The nodule was surgically removed by skin excision, and histologic examination of the specimen confirmed the diagnosis of nodular fascitis. The etiology of the lesion was thought to be a reaction to trauma, in agreement with data reported in the literature. Nodular fasciitis is a benign, non-recurrent lesion; however, its rapid growth and the histologic features it shares with more aggressive tumors such as fibrosarcoma call for careful differential diagnosis. As in our case, a history of previous trauma to the involved area can aid in establishing a correct diagnosis. PMID- 11887081 TI - [Facial nerve paralysis. A case report]. AB - The correction of oral and maxillofacial malformations need invasive and traumatic surgery techniques that may cause damages to nerves. Some lesions related symptoms have been described especially with regard to sensory nerves and, more rarely, also to motor nerves. A case personally observed is described. PMID- 11887083 TI - Scintigraphy with 99mTc-MIBI and echography in the study of primitive hyperparathyroidism. AB - BACKGROUND: The authors analyse the value of the exams preoperative for the identification of the pathological parathyroid glands. METHODS: The authors examined 58 patients affected by primitive hyperparathyroidism (HPTp) who had undergone surgical treatment for primary hyperparathyroidism at the Third Surgical Department of University "La Sapienza" of Rome, in 175 patients affected by primitive hyperparathyroidism observed between January 1970 and June 2000; all patients had undergone echotomography of the neck and 99mTc-MIBI scintigraphy in the diagnostic phase. The histological valuation confirmed the diagnosis by I.P. RESULTS: 99mTc-MIBI scintigraphy demonstrated the pathological glands in 56 cases (96.6%), the echography in 57 cases (98.2%). The comparison of the two exams demonstrated the pathological glands in 100% of the cases. In 45 cases the scintigraphy localized the side (77.6%), and the echography in 41 cases (70.7%). In one case demonstrated I.P. persistent (1.7%) and never one case demonstrated relapsed. CONCLUSIONS: The authors think that the scintigraphy together with the echography show high sensibility to identification the pathological glands. Neck surgical exploration can still give the side good when the diameter of the parathyroids is inferior to 5 mm. PMID- 11887084 TI - Social adjustment in children with Down mental retardation (MRD) and Fragile-X mental retardation (MRX). AB - BACKGROUND: Since adjustment abilities became important in mental retardation (MR) diagnosis, it seemed interesting to study social adjustment in persons with MR Down (RMD) and MR Fragile-X (RMX). These two syndromes are the most common causes of MR of chromosomal origin. To evaluate the influence of temperament insofar as behavior and temperament are concerned in social adjustment, we studied temperamental dimensions (emotionality, activity, sociability and shyness) and social functioning (attention problems and withdrawal). METHODS: Our study group was composed of 35 children with MR; 23 with RMD (F=14) age range 4 to 21, and 12 (F=1) with RMX age ranged from 5 to 19. #Social adjustment was evaluated by two scales: EAS and CBCL. RESULTS: The six evaluated dimensions of adjustment functioning (emotionality, activity, sociability, shyness, attention problems and withdrawal) differ in the two MRD and MRX groups. MRX scores are all higher except for sociability; shyness, attention problems and emotionality show a significant difference. CONCLUSIONS: The RMX group is that one may have more difficulty in social adjustment. This is because they are characterized by hyperactivity, withdrawal, low attention, low social function and high emotionality that are all negative symptoms for a social adjustment. In our study group MRD have higher values in the sociability area and they don't show relevant behavioral disorders and they have got more adaptive abilities. We may hypothesize that this attitude is a part of their genetic structure, and also that the best social adjustment of Down persons may be linked to a better interaction with the environment. PMID- 11887085 TI - Investigation of psychopathological state of patients depending on specific clinical characteristics of physical trauma. AB - BACKGROUND: This article is the retrospective-comparative research of psychopathological reaction of somatically traumatized people. The influence of the outside circumstances of injury (war, job, traffic accident, leisure), injured body part (torso, leg, arm) and different clinical characteristics of injury (surgery, mobile of patient, amputation, acuteness of injury, etc.) are analyzed as potentially factors of the type of the psychical reactions on physical trauma. METHODS: The sample of 70 Serb patients hospitalized in the University Orthopedic Clinic (26 fighters from Bosnia-Herzego-vina and 44 Belgrade residents injured in peace time), was investigated as an experimental group. The control group contained 105 subjects from Belgrade area, 45 of whom had mental trauma experience and 60 had no trauma experience whatsoever. The subjects were tested with the following instruments: Event Effect Scale, PTSS-10 Scale, FHI, Brief Eysenck's Personality Inventory, LEAIQ, GHQ-60, General Questionnaire for Accidents. In the statistical data processing, c2 test, discrimination analysis, and calculation of linear correlation were used. RESULTS: It was determined that the outside circumstances of injury, injured part of the body, amputation, medication and acuteness of the injury did not differentiate any of the psychopathological types of responses analyzed in the research. CONCLUSIONS: Difficult surgery and the immobility of the patient correlated significantly with the high values on the PTSD Scale i.e. the general neuroticism. The seriousness of injury correlated negatively with the intensity of family cohesiveness. PMID- 11887086 TI - Helicobacter pylori influence on gastric acid secretion in duodenal ulcer patients diagnosed for the first time. AB - BACKGROUND: Many experiences have hypothesised that Helicobacter pylori induced hypergastrinemia could lead to an increase of the parietal cell mass and, consequently, of acid secretion. METHODS: The parietal cell mass and maximal acid output have been studied in patients with duodenal ulcer diagnosed for the first time, not due to drugs assumption. In particular, it has been evaluated the parietal cell mass and the acid secretion subdividing duodenal ulcer patients in relation to gastrinemia values (hypergastrinemia and normogastrinemia). RESULTS: The parietal cell mass and the maximal acid output remain high independently of Helicobacter pylori presence. About 60% of the subjects in the Helicobacter pylori positive group show gastrinemia values higher than the average: neither did the study reveal in this group any variations in the parietal cell mass and acid secretion. CONCLUSIONS: It emerges from the results that the mild chronic hypergastrinemia in Helicobacter pylori positive duodenal ulcer is not important enough to induce an increase in parietal cell mass and acid secretion. Therefore, Helicobacter pylori eradication is important in relapse prevention of duodenal ulcer, but not for its repercussions on the gastric secretion. PMID- 11887087 TI - Treatment of heart failure and ascites with ultrafiltration in patients with intractable alcoholic cardiomyopathy. AB - BACKGROUND: In Western countries the excess of alcohol intake causes, secondary, non ischaemic cardiomyopathy and cirrhosis. Frequently, therapy is not effective so ultrafiltration was tried on patients affected, with positive effects on life quality. We tried to verify utility and tolerance to peritoneal ultrafiltration in a group of subjects affected by heart failure secondary to alcoholic cardiomyopathy, refractory to conventional therapy. METHODS: Sixteen patients (14 males, 2 females) with heart failure and ascites affected by alcoholic cardiomyopathy were studied. All subjects were in IV class NYHA (New York Heart Association); ejection fraction (EF) was evaluated by echocardiogram and ascites by abdominal ultrasound. Patients were submitted to clinical exam, body weight, abdominal circumference, diuresis and routine biohumoral exams, electrocardiogram and chest X-ray. Subsequently they underwent intermittent nocturnal peritoneal dialysis with a changing cycle of 6-12 hours per session. After 5 days, subjects were checked through echocardiogram and abdominal ultrasound. RESULTS: The patients mean age was 56.7 +/- 3.2 years. After ultrafiltration, all subjects showed decreased body weight, abdominal circumference and urea; there was an increase of diuresis and Natriuria. Fifteen subjects entered III NYHA class without variation of EF; all of them showed clinical and echographic reduction of ascites. Mean ultrafiltration quantity was 6.084 ml with mean dialysis hours 20; 7.36% of patients had fever that disappeared within 24 hours with antibiotic therapy. All subjects referred to feel well and the mean hospitalization period was of 7 day in spite of the usual 22 days. PMID- 11887088 TI - Food and headache attacks. A comparison of patients with migraine and tension type headache. AB - BACKGROUND: The role of foods as headache precipitants is still matter of debate. Several studies reported that dietary constituents may precipitate migraine attacks. Some authors reported that also tension type headache attacks may be provoked by foods. The purpose of this study was to evaluate the role of foods as headache triggers in both groups of patients. METHODS: We compared the role of foods as headache trigger in patients with migraine and tension type headache. Three hundred and nine patients were involved in the study and divided into six groups: 1) migraine without aura, 2) migraine with aura, 3) episodic tension type headache, 4) chronic tension type headache, 5) migraine without aura associated with episodic tension type headache, 6) migraine without aura associated with chronic tension type headache. RESULTS: Approximately one third of the patients reported susceptibility to certain foods. The percentage of food sensitivity was not significantly different between patients with migraine or tension type headache. The foods more commonly reported as headache triggers were alcoholic drinks, chocolate and cheese. No difference in specific food sensitivity between groups was found. The comparison of food-sensitive with food non-sensitive patients showed no significant difference in the clinical features. CONCLUSIONS: Our study suggests that foods may trigger not only migraine but also tension type headache attacks. PMID- 11887089 TI - Evaluation of antibodies to thymine ROS-modified DNA poly(dT), in patients with immunologic disorders: relationships to anti n-DNA and anti ENA autoantibodies. AB - BACKGROUND: Hydroxyl radical, one of the most potent of all reactive oxygen species, is known to modify adenine and thymine in cellular DNA, producing some modified DNA fragments (ROS-DNA) with different antigenic properties. Despite several in vitro studies about ROS-DNA, data regarding their clinical significance are scanty. The aim of our study was to seek out the presence and clinical significance of the anti poly(dT) auto-antibodies in a group of patients suspected of autoimmune disease. METHODS: We initially evaluated more than 900 consecutive sera of hospitalized patients (range age from 6 to 70 yrs) referred to our laboratory during 18 months. Anti n-DNA, anti-ENA and poly(dT) autoantibodies were performed on 158 ANA positive sera and 28 ANA negative sera from patients strongly suspected of rheumatic diseases or affected by HCV infection. RESULTS: Anti poly (dT) were found in 22 out of 186 sera. As regards the clinical evaluation, 8 patients were affected by SLE, 5 by Scleroderma, 3 by HCV-related chronic hepatitis, 4 by recurrent abortions (without presence of the anti-cardiolipin antibodies and other clinical symptoms). In two patients the ACR criteria and the clinical aspects did not allow a definite diagnosis. Anti Histones were detected in 18 out of 22 poly (dT) positive patients. CONCLUSIONS: Our data suggest that anti poly(dT) autoantibodies are sensitive markers of various autoimmune diseases, but with a minor specificity as compared to anti n DNA for the diagnosis of SLE. PMID- 11887090 TI - Performance and clinical application of a new, fast method for the detection of hydroperoxides in serum. AB - BACKGROUND: Evidence has increased suggesting that a condition of oxidative stress, due to imbalance of oxidative/antioxidant systems, can produce alterations in important biological functions as a consequence of free radicals reactivity towards proteins, lipids, and DNA. Hence the possibility of accurately measuring the oxidative state in a biological system in a simple manner could be of fundamental importance for clinical diagnostics. METHODS: A new, spectrophotometric assay ("d-ROMs test", Diacron s.r.l., Grosseto, Italy), which allows the measurement of reactive oxygen metabolites derivatives, such as hydroperoxides, has been evaluated. RESULTS: The "d-ROMs test" showed good precision, accuracy and linearity. Mean serum d-ROMs above the normal range (344.5 +/- 68.3 U.CARR., mean +/- SD) were detected in a group of heavy drinkers, and well correlated with both g-GT (r=0.44, p<0.04) and MCV (r=0.73, p<0.000), usually considered as biochemical markers of alcohol abuse. CONCLUSIONS: The new test for the measurement of endogenous hydroperoxides proved to be simple, reliable, and cheap. Furthermore, it can be easily applied on automated analyzers. Then it could be used as an early index of oxidative damage, which precedes and partly contributes to degenerative process that affects cell membranes and other lipid containing structures. PMID- 11887091 TI - The advancement of cancer chemoprevention by revision of clinical trial strategies. AB - After a quarter of a century of rapid advances in cancer research, the focus of oncological drug development has shifted from cytotoxic chemotherapy to rationally designed agents that target specific molecules associated with malignant cells or their environment. As a consequence of greater public awareness of health related issues and population screening, the detection of early cancer or premalignant lesions has demonstrated the potential that exists for targeting the same molecules during carcinogenesis. One method of such intervention is chemoprevention, the inhibition, retardation or reversal of carcinogenic processes by chemical means. Tamoxifen and celecoxib have been licensed in the USA for the chemoprevention of breast and colorectal cancers respectively in certain well-defined "at risk" individuals. Unfortunately, many large-scale cancer chemoprevention trials have yielded negative results, and a few may even have caused harm to healthy individuals considered at risk of developing cancer. In this article, the importance of not putting the cart before the horse is argued. In particular, two factors in chemopreventive agent development will be emphasised: phase I/II studies in patients with cancer or patients at risk of second primary tumors and the validation and incorporation of biomarkers of carcinogenesis from the earliest phases of clinical development. This structured approach currently emerging is providing pharmacological and mechanistic data on novel agents which will prove essential in the planning of larger-scale commitments. PMID- 11887092 TI - New concept regarding chest pain due to hypoxia of the internal mammary arteries in more than 1,600 operated patients with cerebral thoracic neurovascular syndrome (CTNVS). AB - In this article we describe the role of compression of the vertebral, subclavian, internal mammary, internal carotid arteries, brachial plexus and coiling and kinking of the vertebral and basilar arteries, the faulty irrigation of blood supply and oxygen of the cerebellum and basal ganglia and other areas of the brain followed by metabolic processes. Among the effects are: a decrease in the secretion of dopamine at the level of the putamen, which produces the symptoms of symptomatic Parkinson's disease, chorea due to chronic transitory faulty blood supply and oxygen to the caudate nucleus, ballism by hypoxia at the level of sub thalamic and thalamus nuclei and athetosis in the lenticular nucleus. This compression is caused by hypertrophy of the anterior scalenus muscles and the cervical ribs at the level of the vertebrae C6-C7; by the sternocleidomastoid at the level of the cervical atlas, by the pectoralis minor muscles and coiling and kinking of the vertebral, basilar and the internal carotid arteries. The decreased blood supply to the cerebellum and basal ganglia is the cause of the cerebral thoracic neuro vascular syndrome (CTNVS) and its neurological complications, among which are ipsilateral paralysis, symptomatic Parkinson's disease, functional Alzheimer's disease multiple sclerosis and others. We are presently engaged in genetic studies to widen our understanding of these illness. PMID- 11887093 TI - The esophageal chest pain. An update for clinicians. AB - Chest pain is a common symptom and even when there is evidence of only minor obstructive coronary artery disease on angiography, it leads to disability and imposes a substantial economic burden on health care system. Gastroesophageal reflux disease (GERD) and esophageal dysmotility disorders are thought to play an important role in the genesis of non-cardiac chest pain. Its pathogenetic mechanism remains unclear. The esophageal origin of the symptom may be identified by an aggressive trial of high-dose antisecretory drugs or an abnormal prolonged ambulatory pH monitoring study. Endoscopy is often normal and less useful in this population than in those with heartburn as presenting symptom. The use of manometry, with provocative testing to evaluate for esophageal motility abnormalities or esophageal sensitivity, allows optimal evaluation of those who do not have GERD. Patients with non-cardiac chest pain of unknown origin should be carefully screened for the occurrence of esophageal disorders but further research is needed to clarify the role of the latter on the pathogenesis of this symptom. PMID- 11887094 TI - Successful treatment of recurrent celiac axis compression syndrome. A case report. AB - The celiac axis compression syndrome is characterized by the clinical triad, epigastric pain, weight loss and postprandial emesis. The aetiology is attributed to intermittent ischaemia of the foregut. The results of three different modalities of treatment, transluminal dilatation, surgical division of the median arcuate ligament, and bypass surgery in a patient with recurrent celiac artery compression syndrome are reviewed. A 62-year-old woman with a previous partial gastrectomy presented with postprandial abdominal pain and marked weight loss. Investigations for gastrointestinal tract pathology were all negative. Angiography revealed compression of the celiac axis and an angioplasty was unsuccessful. The patient underwent surgical division of the median arcuate ligament with complete relief of symptoms, which recurred four months later. Angiography demonstrated a restenosis of the celiac axis. An aorto-celiac artery bypass was done with complete and persistent relief of symptoms at four years follow-up. This is a rare syndrome, which requires predisposing anatomic factors. In this patient, a previous partial gastrectomy with gastrojejunostomy roux-en-y caused disconnection of the pancreatico-duodenal network. Scarring renders ineffective less invasive interventions. Bypass surgery is the treatment of choice and offers durable results. PMID- 11887095 TI - Delayed ANCA positivity in pulmonary-renal syndrome. AB - A 57-year-old man was admitted due to rapidly progressive renal failure and pulmonary edema. Chest X-ray showed a bilateral lung infiltrate, while a normal myocardial contractility was reported by echocardiography. Though initially ANCA were absent, a necrotizing vasculitis with polymorphonuclear leukocyte infiltrate was observed in a kidney biopsy specimen. Renal histology was compatible with microscopic polyangiitis because of necrotizing lesions located at small vessels and at glomeruli that were not crescentic. Corticosteroids and immunosuppressive treatment was played with significant clinical improvement. Six months later, the patient died of a gastrointestinal hemorrhage due to bowel perforation by vasculitic lesion. This time high p-ANCA positivity was detected and at renal histology crescentic glomeruli were observed. PMID- 11887096 TI - Thimerosal: contact (non)allergen of the year. PMID- 11887097 TI - Allergic contact dermatitis from mercury antiseptics and derivatives: study protocol of tolerance to intramuscular injections of thimerosal. AB - BACKGROUND: Mercury derivatives are frequent contact allergens and their cross reactivity is not constant. Thimerosal is an organic mercurial used as an antiseptic and as a preservative in most vaccines. OBJECTIVE: To evaluate cross reactivity, exposure factors, and tolerance to vaccines containing thimerosal in patients sensitized to mercury derivatives. METHODS: DESIGN: Observational study (cross-sectional); PATIENTS: 125 patients were recruited for the study, 72 women and girls and 53 boys and men, average age 18.7 years old, range 3 to 65, with positive patch tests to mercury derivatives and/or thimerosal; INTERVENTIONS: All patients were studied by means of enquiry, patch tests, intradermal tests, and intramuscular challenge with thimerosal. RESULTS: A sensitization to thimerosal was observed in 57 patients. Twenty-four of these 125 patients presented a positive intradermal reaction. Ammoniated mercury seems to be a good marker of mercury sensitization eliciting positive reaction in 78% of all patients and merbromin in 66%. In most cases, (100/125) cross-reactivity was found among mercury derivatives. The intramuscular injection of thimerosal induced a mild local reaction in only 5 patients (4% of the total, 9% of thimerosal positive reactions). Childhood vaccinations, merbromin used as an antiseptic, broken thermometers, and the use of drops were the main sources of exposure. CONCLUSIONS: The majority of the patients showed positive tests to both organic and inorganic mercury derivatives. Vaccination with thimerosal is relatively safe, even for individuals with delayed type hypersensitivity to this chemical, since more than 90% of allergic patients tolerated intramuscular challenge tests with thimerosal. A simplified protocol of patch tests to study mercury derivatives is proposed. It would be advisable to restrict the use of mercurial antiseptics and mercury thermometers. PMID- 11887099 TI - Identification and quantification of para-phenylenediamine in a temporary black henna tattoo. AB - BACKGROUND: Temporary black henna tattoos are very popular as body adornment. Although contact allergy to natural henna is unusual, the inclusion of hair dye, p-phenylenediamine (PPD), increases the risk of contact sensitization. OBJECTIVE: This study was performed to identify the presence and concentration of PPD in a black henna tattoo mixture to which our patient developed contact allergy. METHODS: The presence of PPD in a black henna tattoo mixture, various samples of commercially available henna powders, and several hair dye products was qualitatively and quantitatively detected using high performance liquid chromatography (HPLC). RESULTS: This study demonstrated that PPD was present in the black henna tattoo mixture at a concentration of 15.7%, which is significantly higher than commercial hair dye preparations. CONCLUSION: The presence of PPD in black henna tattoo mixtures in high concentration poses a health hazard and a risk of allergic contact sensitization with potential long term consequences. PMID- 11887098 TI - Bullous phytophotodermatitis associated with high natural concentrations of furanocoumarins in limes. AB - BACKGROUND: Phytophotodermatitis is a phototoxic reaction, occurring in skin exposed to sunlight after contact with plants containing furanocoumarins. Typical reactions are mild, showing erythema with post-inflammatory hyperpigmentation. A 6-year-old boy presented with marked, symmetric, painful erythema and edema of both hands that rapidly developed into dramatic bullae covering the entire dorsum of the hands. The history revealed that the hands had been bathed in lime juice for a prolonged period in the preparation of limeade. OBJECTIVE: This report documents an unusual bullous presentation of phytophotodermatitis resulting from contact with furanocoumarins in local limes. This study was conducted to identify and measure the inciting substances from the rind and pulp of the limes. METHODS: Psoralen, xanthotoxin, bergapten, and isopimpinellin content were measured by gas chromatography and high-pressure liquid chromatography RESULTS: The rind contained 6- to 182-fold concentrations of all furanocoumarins measured when compared with pulp. Bergapten was the most abundant substance in the rind. CONCLUSION: Hydration of the skin during the preparation of limeade combined with increased levels of bergapten in local limes to produce a dramatic bullous reaction. We encourage clinicians to consider the possibility of phytophotodermatitis in severe bullous skin reactions. PMID- 11887100 TI - Patch testing over tattoos. AB - BACKGROUND: There is little, if any, literature regarding placing and reading of patch-tests on tattooed skin. METHOD: A patient whose entire back was covered with multiple tattoos was patch-tested. RESULTS: Multiple positive patch-tests were seen with no apparent reduction in intensity related to tattooed skin. CONCLUSION: If necessary, patch-tests can be placed and read on tattooed skin. PMID- 11887101 TI - A strategy for skin irritation testing. AB - Skin irritation safety testing and risk assessment for new products, and the ingredients they contain, is a critical requirement before market introduction. In the past, much of this skin testing required the use of experimental animals. However, new current best approaches for skin corrosion and skin irritation testing and risk assessment are being defined, obviating the need for animal test methods. Several in vitro skin corrosion test methods have been endorsed after successful validation and are gaining acceptance by regulatory authorities. In vitro test methods for acute, cumulative (repeat exposure), and chronic (prolonged exposure) skin irritation are under development. Though not yet validated, many are being used successfully for testing and risk assessment purposes as documented through an expanding literature. Likewise, a novel acute irritation patch test in human subjects is providing a valid and ethical alternative to animal testing for prediction of chemical skin irritation potential. An array of other human test methods also have been developed and used for the prediction of cumulative/chronic skin irritation and the general skin compatibility of finished products. The development of instrumental methods (e.g., transepidermal water loss, capacitance, and so on) has provided the means for analyzing various biophysical properties of human skin and changes in these properties caused by exposure to irritants. However, these methods do not directly measure skin inflammation. A recently introduced skin surface tape sampling procedure has been shown to detect changes in skin surface cytokine recovery that correlate with inflammatory skin changes associated with chemical irritant exposures or existing dermatitis. It holds promise for more objective quantification of skin irritation events, including subclinical (sensory) irritation, in the future. PMID- 11887102 TI - Occupational contact dermatitis I: incidence and return to work pressures. AB - Since the passage of the United States Occupational Safety and Health Act (OSHA) in 1970, there have been extensive changes in United States workplaces that should have served to enhance the prevention of occupational skin disease (OSD). Analysis of skin diseases reported to OSHA (OSHA recordables) shows that the number of OSDs declined steadily from 1974 to 1983 to about half the previous annual incidence. After 1984, there was a modest resurgence peaking in 1994, with a subsequent decline. A similar but somewhat greater decline in the late 1990s has been observed for occupational respiratory diseases, diseases caused by toxic agents and for poisonings. Likely explanations for the trends in OSD are discussed; the initial decline probably reflected an improvement in workplace conditions, the later resurgence and decline may have been attributable to changes in recording behaviors and in worker's compensation. The decline in recorded OSD since 1996 has been fairly uniform in most major industrial sectors but has been less marked in agriculture, forestry, and fishing so that this sector has replaced manufacturing in recording the highest incidence rate. In 1999, the incidence rate of recorded OSD was 0.49 per 1,000 workers, which appears to grossly under report the true incidence. OSD now constitutes about 10% of all occupational disease cases. Currently, there is increasing emphasis in corporate and occupational medicine on reducing costs and maintaining productivity as well as in preventing occupational injuries and diseases. This is shown by the trend for a greater proportion of workers with occupational conditions to return to modified duty positions rather than to be completely off work. Implications of this phenomena for management of OSD are discussed. PMID- 11887103 TI - Persistent hand eczema in a child. PMID- 11887104 TI - Allergic contact dermatitis from phenylephrine in eyedrops. AB - Phenylephrine is widely used as an ophthalmic drug. However, there are only very few reports on allergic contact dermatitis induced by phenylephrine. In addition, little is known on cross reactivity patterns between the sympathomimetics phenylephrine, epinephrine and ephedrine which share a similar chemical structure. We report on a man who developed allergic contact dermatitis to Neosynerphin POS eyedrops containing phenylephrine hydrochloride. Cross reactivity between phenylephrine, epinephrine and ephedrine was studied by patch testing. Patch tests were performed with the European standard, an ophthalmics and preservatives series, Neosynerphin POS eyedrops, phenylephrine hydrochloride 10% aq., epinephrine and ephedrine (both 1.0 % aq.). Test sites were read after 48, 72 and 168 hours according to the recommendations of the ICDRG. Neosynerphin POS and phenylephrine hydrochloride 10 % aq. gave positive reactions, whereas epinephrine and ephedrine tested negative. Although phenylephrine is an epinephrine analog delayed type hypersensitivity to phenylephrine did not result in cross reactivity with chemically related epinephrine and ephedrine. PMID- 11887105 TI - Allergic contact dermatitis to latanoprost. AB - An 85-year-old male with glaucoma presented with a 1-1/2 year history of tearing; red eyes; and pruritic, edematous, eczematous eyelids. Treatment for presumed ocular rosacea and seborrhea was unhelpful. Patch testing to our standard 64 antigens yielded a positive reaction to Balsam of Peru. Notably, benzalkonium chloride, thimerosal, and other preservatives elicited negative reactions. Repeat open application testing elicited positive results to Xalatan (latanoprost) 0.005% ophthalmic solution (Pharmacia & Upjohn, Kalamazoo, MI). A second session of patch tests to 10 personal products, in addition to Xalatan 0.005% solution and the Xalatan vehicle (provided by the manufacturer), elicited a strong positive reaction only to the full preparation of Xalatan 0.005% solution. This report describes the first known case of ACD to latanoprost, a new prostaglandin analog that is widely prescribed for treatment of glaucoma. PMID- 11887106 TI - Contact dermatitis in a woodworker. AB - Woods are capable of causing allergic or irritant contact dermatitis which typically occurs on the exposed areas of the arms, face, and neck. The allergens found in woods include quinones, stilbenes, phenols, and terpenes. We report an 84-year-old woodworker who developed allergic contact dermatitis from Bolivian rosewood and Cocobolo wood. The patient was patch-tested using the North American Contact Dermatitis Group Standard Tray; 2,6 dimethoxyl 1,4 benzoquinone; and wood that he had been exposed to on a regular basis. Positive patch test reactions occurred to methyldibromo glutaronitrile phenoxyethanol, sodium gold thiosulfate, Bolivian rosewood, and Cocobolo wood. Allergens found in Bolivian rosewood and Cocobolo wood caused this patient's chronic dermatitis, which cleared when he avoided these woods. PMID- 11887107 TI - Piriformis syndrome: diagnosis, treatment, and outcome--a 10-year study. AB - OBJECTIVES: To validate an operational definition of piriformis syndrome based on prolongation of the H-reflex with hip flexion, adduction, and internal rotation (FAIR) and to assess efficacy of conservative therapy and surgery to relieve symptoms and reduce disability. DESIGN: Before-after trial of cohorts identified by operational definition. SETTING: Outpatient departments of 2 hospitals and 4 physicians' offices. Surgery performed at 3 hospitals. PATIENTS: Consecutive sample of 918 patients (1014 legs) with follow-up on 733. INTERVENTION: Patients with significant (3 standard deviations [SDs]) FAIR tests received injection, physical therapy, and serially reported pain and disability assessments. Forty three patients (6.47%) had surgery. MAIN OUTCOME MEASURES: Likert pain scale. Subjective estimates of disablement in activities of daily living and instrumental activities of daily living. RESULTS: At 3 SDs, the FAIR test had sensitivity and specificity of.881 and.832, respectively. Seventy-nine percent (514/655) of FAIR test positive (FTP) patients improved 50% or more from injection and physical therapy at a mean follow-up of 10.2 months. Average improvement was 71.1%. Of 385 FTP patients with disability data, mean disability fell from 35.37% prestudy (SD =.2275) to 12.96% poststudy (SD =.1752), a 62.8% improvement. Twenty-eight surgical FTP patients (68.8%) showed 50% or greater improvement; mean improvement was 68% at a mean follow-up of 16 months. Surgery reduced the mean FAIR test to 1.35 +/- 2.17 months postoperatively. FTP patients generally improved 10% to 15% more than others after conservative treatment. CONCLUSIONS: The FAIR test correlates well with a working definition of piriformis syndrome and is a better predictor of successful physical therapy and surgery than the working definition. The FAIR test, coupled with injection and physical therapy and/or surgery, appears to be effective means to diagnose and treat piriformis syndrome. PMID- 11887108 TI - Mixed median nerve forearm conduction velocity in the presence of focal compression neuropathy at the wrist versus peripheral neuropathy. AB - OBJECTIVES: To establish normal values for the conduction velocity of the mixed median nerve in the forearm, and to determine the use of mixed median nerve conduction velocity (NCV) studies across the forearm in the differential diagnosis of carpal tunnel syndrome (CTS), peripheral neuropathy, and CTS with peripheral neuropathy. DESIGN: Criterion standard. Mixed median NCV studies across the forearm were added to routine CTS investigational protocols. SETTING: University outpatient setting, rural referral base. PARTICIPANTS: Thirty healthy volunteers and 60 patients referred with symptoms and signs suggestive of CTS and/or peripheral neuropathy. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Motor sensory and mixed median nerve conduction velocity across the forearm. RESULTS: The mean mixed median NCV across the forearm was 64.5 m/s in healthy subjects and subjects with CTS. The mean mixed NCV in subjects with peripheral neuropathy was 54 m/s, even in the presence of CTS. CONCLUSION: Mixed median NCV in the forearm added to the standard protocols is helpful in differentiating the diagnosis of CTS from peripheral neuropathy and CTS with peripheral neuropathy. PMID- 11887110 TI - Chronic medial and lateral epicondylitis: a comparison of pain, disability, and function. AB - OBJECTIVES: To investigate pain, disability, and muscle function of the arm in medial epicondylitis and to compare the results with those in chronic lateral epicondylitis. DESIGN: Cross-sectional, case-control study. SETTING: University hospital clinic admitting chronic hand patients. PARTICIPANTS: Twenty-five patients with chronic unilateral medial epicondylitis and 25 age- and gender matched patients with chronic unilateral lateral epicondylitis. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Pain drawing (PD) and a pain questionnaire with 7 items of pain and disability on visual analog scale (VAS). Dolorimeter measurements of pressure pain thresholds (PPT) on 3 defined cubital points. The isometric grip strength and isokinetic performance of wrist and forearm at a radial velocity of 90 degrees/s. RESULTS: Patients with medial epicondylitis had significantly less pain under strain (6.7 vs 7.9cm on VAS, P =.03) and a smaller PD (1.9 vs 2.5, P =.02) than patients with lateral epicondylitis. The PPTs of medial epicondyles were 54% (P =.0000) lower in medial epicondylitis. In lateral epicondylitis, all 3 cubital points showed significant decreases in PPTs. The isometric grip strength (mean and maximal) decreased by 6.2% and 3.6%, compared with the patients' healthy arms (P =.03,.16) and by 11.4% and 8.9% (P =.008,.02), respectively, compared with the expected value; in lateral epicondylitis, the grip strength decreased by 11.8% and 10.6% (P =.005,.01) and by 15% and 14% (P =.003,.007), respectively, when compared with the expected grip strength. Peak torque and produced work in wrist flexion were significantly reduced by 13% and 17% (P =.005,.0001), respectively, in both diseases. In lateral epicondylitis, supination and pronation were also reduced by 10% and 15% (P =.03). CONCLUSIONS: In chronic medial epicondylitis, muscle function and pain measures showed a lesser impaired function of the arm than in chronic lateral epicondylitis. The results may be useful in rehabilitation and treatment of epicondylitis. PMID- 11887109 TI - Concentric versus combined concentric-eccentric isokinetic training: effects on functional capacity and symptoms in patients with osteoarthrosis of the knee. AB - OBJECTIVE: To compare the effects of concentric and coupled concentric-eccentric isokinetic resistance training on functional capacity and symptoms of patients with osteoarthrosis (OA) of both knees. DESIGN: Repeated measures. SETTING: A university exercise physiology laboratory. PARTICIPANTS: Twenty-three volunteers, ages 41 to 75 years. Patients were randomly assigned to 3 groups: concentric (CON, n = 9), concentric-eccentric (CON-ECC, n = 8), and nontreatment (NONTX, n = 6). INTERVENTIONS: The CON group performed 12 concentric contractions of each knee extensor and flexor muscles; the CON-ECC group performed 6 concentric and 6 eccentric contractions of each knee muscle group by using a spectrum of angular velocities ranging from 30 degrees/s to 180 degrees/s with 30 degrees/s intervals, for both sides, 3 days a week for 8 weeks. MAIN OUTCOME MEASURES: Functional capacity (rising from a chair, walking, stair climbing and descending) and pain during rest and activities, peak torque, and cross-sectional area (CSA) of knee muscle groups of subjects were tested before and after training. RESULTS: Both training groups showed marked decreases (P <.001) in pain scores and increases (P <.001) in functional capacity together with increases (P <.05--.01) in peak torque and CSA of knee muscles. However, the NONTX group did not display these marked changes after the 8-week period. The results also indicated that concentric-eccentric training has a greater influence on functional capacity, especially stair climbing and descending, than that of concentric training when compared with NONTX group; however, the improvements in pain measurements were better in the CON group compared with the CON-ECC group after the training. CONCLUSIONS: The results suggest that with the isokinetic resistance training used in this study, it is possible to improve functional capacity and decrease pain in patients with OA of the knee. The results also showed that extensive training involving a high number of repetitions and eccentric contractions was safe, effective, and well tolerated for the patients with knee OA. PMID- 11887112 TI - The incidence and consequences of falls in stroke patients during inpatient rehabilitation: factors associated with high risk. AB - OBJECTIVES: To determine the incidence of falls on a stroke rehabilitation unit; to assess the frequency and nature of injuries; and to identify risk factors predictive of falls, functional outcomes, and impairments. DESIGN: Retrospective cohort study. SETTING: An inpatient stroke rehabilitation unit. PARTICIPANTS: Two hundred thirty-eight consecutive stroke patient admissions. INTERVENTIONS: Incident reports completed on patients who experienced a fall while on the unit were reviewed and resultant injuries categorized (abrasions, lacerations, fractures). MAIN OUTCOME MEASURES: Stroke impairments and admission functional assessments, FIM instrument, Berg Balance Scale (BBS), and Chedoke-McMaster (CM) Stroke Impairment Inventory of fallers were compared with nonfallers. RESULTS: Of the 238 patients, 88 (37%) experienced at least 1 fall, and almost half of these (45 patients [19%]) experienced at least 2 falls. A total of 180 falls were reported over the 5-year period. Of the 180 reported falls, 33% occurred when patients were using their wheelchairs. Injuries occurred in 22% of the reported falls. These consisted of contusions (49%) and abrasions (41%), primarily of the upper (30.8%) and lower (25.6%) extremities. Only 1 fracture was reported. Fallers tended to have lower admission BBS scores (50% of patients with a score <30 fell vs 18% with a score >30, P <.01) and a lower score on the admission arm, leg, and foot components of the CM (P <.05). Patients who fell were also more likely to be apraxic (P <.014) and suffer from cognitive deficits (P <.01). Repeat fallers had lower admission FIM scores (P <.01) when compared with nonfallers. CONCLUSION: Although patients undergoing stroke rehabilitation experienced a significant number of falls, the incidence of serious injury was small. Patients who experienced at least 1 fall had significantly lower BBS, FIM, and CM arm, leg, and foot scores compared with nonfallers. These data suggest that groups of stroke patients who are at risk for falls within the rehabilitation setting can be identified by using a variety of impairment and functional assessments. This information may be potentially useful for designing interventions directed at reducing fall frequency among stroke survivors. PMID- 11887111 TI - Reliability of lower extremity strength measures in persons with chronic stroke. AB - OBJECTIVE: To determine the reliability of isokinetic concentric strength measures of both the hemiparetic and noninvolved limbs for flexion and extension motions of the hip, knee, and ankle joints in individuals who have had a stroke. DESIGN: Test-retest, repeated-measures intraobserver reliability design. SETTING: Tertiary rehabilitation center. PARTICIPANTS: Twenty community-dwelling individuals who have had a stroke, with motor deficits ranging from 3 to 6 on the Chedoke-McMaster Stroke Assessment; volunteer sample. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Peak torque and average torque (ie, mean over the range of motion tested) from an ensemble-averaged (3 trials) torque-angle curve during isokinetic concentric extension and flexion movements of the ankle, knee, and hip. RESULTS: Although peak and average torque were significantly less for the hemiparetic limb compared with the noninvolved limb, the intraclass correlation coefficients (ICCs) between the 2 test sessions were high (.95-.99 for peak torque,.88-.98 for average torque) for both limbs for all 3 joints. However, there was a learning effect, as observed by the slightly greater values attained from the second test session. CONCLUSIONS: Peak and average isokinetic torque can be used to assess reliably lower extremity strength in persons with chronic stroke. Practice sessions may be required before the actual test to reduce the effect of learning. PMID- 11887113 TI - Determinants of driving after stroke. AB - OBJECTIVE: To identify variables that best predict a team's decision of driving ability in stroke patients from a predriving assessment. DESIGN: Retrospective study of a 2-year predriving evaluation. SETTING: Belgian Institute for Road Safety. PARTICIPANTS: One hundred four patients with sequelae of first stroke. INTERVENTIONS: Predriving assessments and road test. MAIN OUTCOME MEASURES: The suitability to resume driving based on a team decision and performance in the road test. RESULTS: Forty-one patients (39.4%) were judged suitable, 45 (43.3%) not immediately suitable, and 18 (17.3%) not suitable to drive. Correlation coefficients and comparisons between groups revealed that most variables had significant individual relationships with the team decision and performance on the road test. After logistic regression analysis, side of lesion, kinetic vision, visual scanning, and a road test led to the best model in predicting the team decision (R(2) =.53). The road test was the most important determinant (R(2) =.42). Multiple regression analysis showed that the combination of acuity of left and right eyes and the figure of Rey was the best subset to predict the road test (R(2) =.28). CONCLUSION: The predictive accuracy of the team's decision is limited, and the road test is even lower. Inclusion of more real-road-related tests in the predriving assessment is necessary. PMID- 11887114 TI - Mini-Mental State Examination, cognitive FIM instrument, and the Loewenstein Occupational Therapy Cognitive Assessment: relation to functional outcome of stroke patients. AB - OBJECTIVES: To compare 3 cognitive tests, used on admission, for predicting discharge functional outcome and to assess the efficacy of these tests in predicting functional outcome at discharge in stroke patients undergoing rehabilitation. DESIGN: Cohort study. SETTING: Geriatric rehabilitation department of a tertiary care hospital in Israel. PATIENTS: Sixty-six patients undergoing acute inpatient comprehensive rehabilitation after first clinical stroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASUREMENTS: Cognitive status was assessed with the Loewenstein Occupational Therapy Cognitive Assessment (LOTCA), the Mini-Mental State Examination (MMSE), and the cognitive subscale of the FIM instrument. The FIM motor subscale was used to assess functional outcome status. Functional gain was determined by the motor FIM gain (efficacy), and the relative (to potential) functional gain was determined by the Montebello Rehabilitation Factor Score. Efficiency was calculated by efficacy divided by the length of hospital stay. RESULTS: A significant increase in total FIM scores (34.8 points) occurred during rehabilitation mainly because of improvement in motor functioning (31.5 points). Significant improvement in global cognitive status was documented by all 3 tests. Intertest correlation coefficients ranged between.47 and.67. The LOTCA showed somewhat higher correlation coefficients with most of the parameters of functional motor outcomes. Correlation between the MMSE and FIM cognitive subscale and these outcome parameters were nearly identical. CONCLUSION: The LOTCA is slightly better than the MMSE and the FIM cognitive subscale in predicting functional status change after stroke rehabilitation but it is a time-consuming and exhausting instrument to use. The FIM cognitive subscale requires a better overall understanding of the patient's situation at time of administration and therefore is less convenient for the initial assessment. The similar correlation of all 3 tests with functional outcomes and the simplicity of administration of the MMSE suggests its use in the initial assessment of stroke patients. PMID- 11887115 TI - Excess risk of bladder cancer in spinal cord injury: evidence for an association between indwelling catheter use and bladder cancer. AB - OBJECTIVES: To evaluate whether the risk of bladder cancer is greater in individuals with spinal cord injury (SCI) than in the general population and whether indwelling catheter (IDC) use is a significant independent risk factor for bladder cancer. DESIGN: Historical cohort study in which subjects with SCI were stratified according to bladder management method and followed for the development of bladder cancer. SETTING: A large rehabilitation hospital in the Spinal Cord Injury Model Systems. PARTICIPANTS: A total of 3670 patients with SCI who were evaluated for bladder cancer on at least 1 occasion by cystoscopy over a period of 1 to 47 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Bladder cancer occurring after SCI determined by diagnosis at our facility, by subject report, or by report of next of kin. RESULTS: Twenty-one cases of bladder cancer were found in the 3670 study participants. The risk of bladder cancer for subjects with SCI using IDC is 77 per 100,000 person-years, corresponding to an age- and gender-adjusted standardized morbidity ratio (SMR) of 25.4 (95% confidence interval [CI], 14.0--41.9) when compared with the general population. After controlling for age at injury, gender, level and completeness of SCI, history of bladder calculi, and smoking, those using solely IDC had a significantly greater risk of bladder cancer (relative risk [RR] = 4.9; 95% CI, 1.3--13.8) than those using nonindwelling methods. Mortality caused by bladder cancer in individuals with SCI was significantly greater than that of the US population (SMR = 70.6; 95% CI, 36.9--123.3). CONCLUSIONS: Bladder cancer risk and mortality are heightened in SCI compared with the general population. IDC is a significant independent risk factor for the increased risk of and mortality caused by bladder cancer in the SCI population. PMID- 11887116 TI - Long-term mental health of men after a first acute myocardial infarction. AB - OBJECTIVE: To evaluate the differential and independent impact of sociodemographic, medical, and psychologic variables assessed at hospital discharge on patients' short- and long-term mental health. DESIGN: Longitudinal study. SETTING: Eight medical centers in central Israel. PARTICIPANTS: Male Israeli patients (N = 209; age range: 30-65 y) with documented first acute myocardial infarction (AMI). INTERVENTION: Subjects were interviewed 3 times, once (T1) before hospital discharge, a second time (T2) at 3 to 6 months after discharge, and a third time (T3) at 5 years post-AMI. Sociodemographic, medical, and psychologic data were elicited at the first interview and completed with medical information in the files. Psychologic well-being and psychologic distress were evaluated by the Mental Health Inventory at the second and third interviews. These 2 outcome variables were compared with normative community data on these aspects of mental health. MAIN OUTCOME MEASURES: Hierarchical regression was used to examine the relation between the independent variables, sociodemographic, medical, and psychologic variables, and the dependent variables, psychologic well being and psychologic distress, at T2 and T3. RESULTS: Depression, perceived health, sense of coherence, social support, and educational level at discharge predicted aspects of mental health 3 to 6 months and 5 years post-AMI. However, only psychologic distress differentiated between the research participants and the normative community sample of men. CONCLUSIONS: A first episode of AMI appears to increase psychologic distress more than it decreases psychologic well being both 3 to 6 months and 5 years post-AMI. Educational level and sense of coherence may serve as protective factors, whereas depression may foster vulnerability to distress and impaired psychologic well-being. PMID- 11887117 TI - Characterization of tissue resistance during a dorsally directed translational mobilization of the glenohumeral joint. AB - OBJECTIVES: To quantify forces applied by therapists during dorsal glide translational mobilization of the glenohumeral joint, to determine the relationship of tissue resistance to the load-displacement relation of the glenohumeral joint, and to determine the safety of the forces applied by the therapists during dorsal glide translational mobilization. DESIGN: A fresh cadaver shoulder specimen mounted on a 6-axis load cell was used to register forces applied by therapists during dorsal glide translational mobilization of the glenohumeral joint in a test-retest pattern. SETTING: Biomechanics laboratory. PARTICIPANTS: Twelve experienced orthopedic physical therapists. INTERVENTION: Not applicable. MAIN OUTCOME MEASURES: Forces exerted by therapists during passive dorsal glide translational mobilization in the loose-packed position and in the end range of abduction, with different grades of movements. The movements did not include any manipulation or thrust-type procedures. Simulated dorsal glide procedures were performed by the material testing system to construct the load-displacement curve of the glenohumeral specimen. The corresponding locations of the forces applied by therapists were interpolated and plotted on the load-displacement curve. RESULTS: The peak force values measured during mobilization were characterized by large intertherapist variability: coefficients of variation ranged from 40.97% to 77.49%. Test-retest reliability for intrasession measures was high (ICC(2,1) range,.90-.94); intersession reliability was poor (ICC(2,1) range,.01-.54). The mean forces ranged from 18.36 to 38.76N. When interpolated to the load-displacement curve, the mean peak forces obtained fell mostly in the toe and the linear elastic regions of the load displacement curve. CONCLUSION: Force parameters measured during dorsal glide mobilization were characterized by large intertherapist variability with high intrasession and poor intersession test-retest reliability. The mobilization forces applied by experienced orthopedic physical therapists fall safely in the toe and the linear elastic regions of the load-displacement curve. PMID- 11887118 TI - Effect of scapular protraction and retraction on isometric shoulder elevation strength. AB - OBJECTIVES: To examine the effect of scapular protraction (SP) and scapular retraction (SR) on isometric shoulder elevation strength measured in the sagittal plane and to test the hypothesis that strength would be significantly reduced when tested in the SP position relative to the neutral resting scapular position (SN). DESIGN: Prospective before-after trial. SETTING: Multidisciplinary sports medicine center. PARTICIPANTS: Ten healthy volunteers (5 men, 5 women) ages 26 to 43 years recruited from the Mayo Clinic, Rochester, MN. INTERVENTIONS: Subjects completed 3 maximal isometric shoulder elevation contractions at 90 degrees of sagittal plane elevation in the SN, SP, and SR positions. The order of scapular positions was varied to minimize fatigue effects. Mean isometric strength values were compared by using Student t tests. MAIN OUTCOME MEASURES: Isometric shoulder elevation strength for the 3 scapular positions. RESULTS: Isometric strength was significantly lower for the SP position compared with the SN position (8.5 +/- 3.4 kg vs 11.1 +/- 4.0 kg, P <.0005) and for the SR position relative to the SN position (7.8 +/- 3.3 kg vs 11.1 +/- 4.0 kg, P <.00003). Strength values did not differ between the SP and SR positions (P =.38). CONCLUSIONS: Movement of the scapula into a protracted or retracted position results in a statistically significant reduction in isometric shoulder elevation strength as measured in this study. Further research is warranted to examine the relationship between scapular position and shoulder muscle function. PMID- 11887119 TI - Impact of physician reminders on the use of influenza vaccinations: a randomized trial. AB - OBJECTIVE: To analyze the impact of mailed physician reminders to immunize their patients. DESIGN: Randomized trial involving Washington State physiatrists participating in the Medicare program. In 1997, all physiatrists in the state were separated into solo or group practice. Solo physicians and group practices were then separately randomized to receive 4 monthly reminders to have their patients immunized. In 1998, the intervention and control groups were switched. SETTING: The state of Washington. PATIENTS: A total of 4300 Medicare outpatients seen in Washington State in 1997 and 4025 patients in 1998. INTERVENTION: Repeated mailer. MAIN OUTCOME MEASURE: By using multivariate analysis, Medicare billing data was analyzed to determine the impact of the physician reminders on influenza vaccination rates. RESULTS: Among solo practitioners, patients whose physiatrist received the reminder letters in 1998 were 34% more likely (adjusted relative risk [RR] = 1.34; 95% confidence interval [CI],.96-1.88) to have a vaccination billing. Among group practitioners, those patients whose physiatrist received the reminder letters in 1997 were 26% more likely (RR = 1.26; 95% CI,.98 1.60) to have a vaccination billing. These differences, however, were not statistically significant. The adjusted RRs for the remaining intervention groups, solo practitioners in 1997 (RR =.89; 95% CI,.63-1.26), and group practitioners in 1998 (RR = 1.00; 95% CI,.73-1.36), revealed no increase in vaccination billings for patients whose physiatrist received the intervention. CONCLUSIONS: Repeated physician reminders did not increase the vaccination rate of Washington State Medicare patients who were seen by physiatrists in 1997 and 1998. These results were consistent whether the physiatrists were in solo or group practice. Other methods should be considered to improve the primary care delivered to this Medicare population. PMID- 11887120 TI - French translation and validation of 3 functional disability scales for neck pain. AB - OBJECTIVE: To translate and assess the reliability and the construct validity of 3 functional disability scales for neck pain. DESIGN: Reliability and validity study. SETTING: Tertiary care teaching hospital and outpatient clinic. PARTICIPANTS: One hundred one patients (mean age, 49 y). INTERVENTION: French translations were obtained by using the "translation-backward translation" method. Adaptations were made after a pilot study. MAIN OUTCOME MEASURES: Impairment outcome measures (visual analog scale [VAS] pain, neck range of motion, morning stiffness, score of neck sensitivity, radiologic score of Kellgren) and patients' perceived handicap (VAS) were recorded at the baseline visit. Three functional disability scales (Neck Disability Index [NDI], Neck Pain and Disability Scale [NPDS], Northwick Park Neck Pain Questionnaire [NPQ]) were recorded twice, at baseline visit and 24 hours later. Reliability was assessed by using the intraclass correlation coefficient (ICC) and the Bland and Altman method. Construct (convergent and divergent) validity was investigated by using the Spearman rank correlation coefficient and a factor analysis was performed. RESULTS: Test-retest was excellent for the NPDS and NDI (ICC =.91,.93, respectively) and good for the NPQ (ICC =.84). The Bland and Altman method showed no systematic trend. Expected convergent and divergent validity were observed only for the NPDS; 3 main factors were extracted by factor analysis and explained 78% of the cumulative variance. CONCLUSION: The 3 translated scales are valid, but the NPDS seems to have the best construct validity. PMID- 11887121 TI - Use of item response theory to link 3 modules of functional status items from the Asset and Health Dynamics Among the Oldest Old study. AB - OBJECTIVE: To link, by using item response theory (IRT), 3 modules of functional status items in the Asset and Health Dynamics Among the Oldest Old (AHEAD) study. DESIGN: Secondary data analysis of the functional status items in the AHEAD study. In that study, participants completed a common set of functional status items and were randomly assigned to complete 1 of 2 modules containing different functional status items. SETTING: A nationally representative panel study of elderly. PARTICIPANTS: US baseline data from 4655 respondents. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Nineteen common items and 2 sets of supplemental items that measure functional status. RESULTS: A 2-parameter model for dichotomous items was used for linking the 3 modules of items. By using this model, both sets of supplemental items were successfully linked to the common items, allowing the placement of all items on the same underlying measure of ability. The small-muscle instrumental activities of daily living were the easiest of all the items for respondents to perform. The item on walking from the Longitudinal Study of Aging was the most difficult for respondents to perform. CONCLUSIONS: IRT-based linking methods were a useful way to overcome test dependency and to place items on a common metric even if different respondents answer different sets of items. Numerous important design features can degrade linking results and should be attended to in future linking studies. PMID- 11887122 TI - Assessment of the impact of pain and impairments associated with spinal cord injuries. AB - OBJECTIVES: To determine the adequacy of the Multidimensional Pain Inventory (MPI) for assessing pain impact after spinal cord injury (SCI) and to determine whether the impact of pain can be separated from other consequences of SCI. DESIGN: Postal survey. SETTING: General community. PARTICIPANTS: Of the 159 subjects contacted who experienced chronic pain, 120 (75.5%) participated. INTERVENTIONS: Subjects were mailed the original MPI and a set of additional items specific to SCI. MAIN OUTCOME MEASURE: The MPI. RESULTS: Confirmatory (CFA) and exploratory factor analyses were performed for each section of the MPI. Elimination of several items, including those related to work in section 1 (pain impact), improved the goodness-of-fit index (GFI). A CFA for section 2 (response of significant other) resulted in acceptable GFI after 2 items were deleted. Decrease in activity levels (section 3) because of other consequences of injury was significantly greater after tetraplegia than after paraplegia. In contrast, pain-related reduction in activities was not associated with injury level. Although other consequences of SCI may have greater impact on activities than pain, severe pain is likely to affect activity levels significantly. CONCLUSION: The MPI appears to be appropriate for use in a SCI population when modified to eliminate questions related to work and to supplement the activity scale with items addressing decreased activity levels due to pain. PMID- 11887123 TI - More than meets the eye: how examiner training affects the reliability of the MacNeill-Lichtenberg decision tree in geriatric rehabilitation patients. AB - OBJECTIVE: To examine the effect of examiner training on the test-retest reliability of the MacNeill-Lichtenberg Decision Tree (MLDT), a tool for guiding clinicians' decision making for referrals for mental health problems. DESIGN: Correlational analyses and chi-squares were used to examine the influence of demographic variables on MLDT performance and the test-retest reliability of its cognitive and affective components. SETTING: Rehabilitation unit of a large, freestanding, urban hospital. PARTICIPANTS: In study 1, 39 older, medical rehabilitation patients consecutively referred to the neuropsychology service. In study 2, 57 older, consecutively admitted medical rehabilitation patients. INTERVENTIONS: In study 1, patients underwent testing with the MLDT by a novice examiner. In study 2, patients were tested by trained examiners. Both sets of results were compared with those obtained by experienced examiners. MAIN OUTCOME MEASURES: The temporal stability of participants' performance on the cognitive and affective components of the MLDT was compared between the 2 studies. RESULTS: Training was associated with high test-retest reliability on both the cognitive and affective components. A lack of training was associated with reduced reliability in depression screening. CONCLUSION: These findings support the use of the MLDT as a mental health triage tool for older adults in inpatient medical settings. Its use is dependent on training and accurate administration. PMID- 11887124 TI - Effect of neck pain on verticality perception: a cohort study. AB - OBJECTIVE: To use the Rod-and-Frame Test (RFT) as a quantification of the perception of verticality in subjects with and without neck pain. DESIGN: Cohort study comparing perception of verticality in symptomatic subjects with neck pain versus a control group. SETTING: Both groups were selected from 2 urban chiropractic offices treating typical neuromusculoskeletal conditions from the general community in Canada. PATIENTS: Nineteen subjects (11 women, 8 men) with uncomplicated mechanical neck pain and 17 (7 women, 10 men) asymptomatic subjects. INTERVENTION: The RFT offers a noninvasive method of measuring spatial orientation or the perception of verticality. Studies of the RFT indicate that performance is reliable. The RFT requires subjects to set a luminescent rod to the true vertical in the presence and absence of a luminescent background frame. MAIN OUTCOME MEASURE: The amount of rotation was measured and recorded by a dial on the back of the device. RESULTS: Two-way analysis of variance showed statistically significant differences in judging vertical between symptomatic and asymptomatic subjects. Unpaired t tests for each test situation and the Tukey post hoc test showed statistical differences for both groups. CONCLUSION: There may be a direct connection between the structures that provide internal cues for the body's ability to sense verticality and nociceptive influences affecting the afference of these structures. The overshoot of the symptomatic group could indicate the search for additional proprioceptive information. PMID- 11887125 TI - Pediatric physiatry in 2000: a survey of practitioners and training programs. AB - OBJECTIVES: To assess the current status and future prospects of the field of pediatric rehabilitation medicine (PRM) physiatry by detailing the demographics, training, research interests, and other characteristics of physicians currently practicing in that field and to determine the availability of training programs in PRM. DESIGN: A printed survey of members of the American Academy of Physical Medicine and Rehabilitation Pediatric Rehabilitation Special Interest Group (PRSIG) and a separate questionnaire directed to departments of physical medicine and rehabilitation (PM&R) concerning their PRM training programs. SETTING: Not applicable. PARTICIPANTS: PRSIG members and PM&R training programs listed by the American Board of Physical Medicine and Rehabilitation. INTERVENTION: Between July 1998 and April 2000, a survey was sent to PRSIG members, with follow-up mailings to nonresponding members. Between April and July 2000, a survey on PRM training practices was sent to 82 PM&R departments with fax and telephone follow up to nonresponders and to those departments that had discontinued their training program. MAIN OUTCOME MEASURES: Instrument measures of physicians practicing PRM, including demographics, geographic distribution, practice sites, training, academic participation, special interests, and research interests along with willingness to participate in collaborative research, association in other organizations, and communication preferences. Data on training requirements as well as availability of PRM fellowships and combined pediatric and PM&R residencies indicate that the number and scope of these training opportunities are declining. RESULTS: There is little uniformity in the amount of exposure to pediatric rehabilitation required by PM&R residency training programs. There are discrepancies in the reported numbers of PRM fellowships and/or combined pediatric and PRM training programs. Projecting the number of physicians who will be practicing in this subspecialty in the future is difficult because there are no reliable data about the number of graduates or trainees in the field. CONCLUSIONS: The rehabilitation needs of children are met by physiatrists with specialized pediatric training. Our survey provided a demographic overview of the PRSIG membership. Given the decline in PRM training programs, it is imperative that the remaining programs be strengthened through communication and organization among physiatrists who practice PRM. We believe that a national database and an interactive Web site are feasible means with which to facilitate this goal. PMID- 11887126 TI - Frontal impairment and confabulation after herpes simplex encephalitis: A case report. AB - We describe the rehabilitation training of a 53-year-old woman with severe confabulatory and dysexecutive syndrome, as well as memory impairment, after herpes simplex encephalitis (HSE). Secondary narcolepsy was also present. Neuropsychologic deficits were detailed through an extensive examination, and specific techniques were used to improve performances in each defective cognitive domain. Improvement of vigilance and attention was reached through appropriate and timed periods of rest, along with attentional tasks of growing difficulty. Different external aids were used to reduce temporal disorientation, to contrast confabulation and inertia, and to overcome memory deficits in everyday life. Their independent use by the patient was implemented through cues that were progressively reduced. Treatment also focused on planning, categorization, and topographic orientation. The patient's family gave constant support during rehabilitation and provided informal training after discharge. The patient was able to regain independence in everyday life at home. PMID- 11887127 TI - Sudden onset of cervical spondylotic myelopathy during sleep: a case report. AB - Cervical spondylotic myelopathy is a common cause of compressive spinal cord dysfunction. The typical course involves either a gradual or an episodic increase in symptoms and neurologic deficits, with impairment evolving over a period of months to years. Acute neurologic deterioration in conjunction with cervical spondylosis has been described almost exclusively in traumatic situations such as disk herniation. We report a case of an acute, nontraumatic onset of tetraplegia in association with cervical spondylosis. A 56-year-old man developed tetraplegia during a 1-hour nap, with loss of volitional control of his extremities, impaired sensation below the C3 dermatome, and increased muscle tone. Magnetic resonance imaging of the cervical spine revealed canal stenosis and increased T2 signal within the cord. This case report describes the rehabilitation course for this patient and reviews the clinical spectrum of onset and progression of cervical spondylotic myelopathy. PMID- 11887128 TI - Occult maxillary sinusitis as a cause of fever in tetraplegia: 2 case reports. AB - Common causes of fever in tetraplegia include urinary tract infection, respiratory complications, bacteremia, impaired autoregulation, deep vein thrombosis, osteomyelitis, drug fever, and intra-abdominal abscess. We report 2 acute tetraplegic patients who presented with fever of unknown origin. After extensive work-up, they were diagnosed with occult maxillary sinusitis. A search of current literature revealed no reports of sinusitis as a potential source of fever in recently spinal cord--injured patients. Patients with tetraplegia, especially in the acute phase of spinal cord injury, often undergo nasotracheal intubation or nasogastric tube placement, which may result in mucosal irritation and nasal congestion. All of the previously mentioned factors, in combination with poor sinus drainage related to supine position, predispose them to developing maxillary sinusitis. The 2 consecutive cases show the importance of occult sinusitis in the differential diagnosis of fever in patients with tetraplegia. PMID- 11887129 TI - Form may be as important as function in orthotic acceptance: a case report. AB - Orthotic prescription offers interesting insights into patient compliance with what a physician or therapist recommends. For example, when a deficit is severe, an orthotic device or gait aid as large and intrusive as a cane or ankle-foot orthosis is usually well accepted. However, an orthosis, even if beneficial, may be discarded if it is not essential to performing daily activities, is uncomfortable, or if the patient feels it highlights his/her disability. Nevertheless, it is sometimes possible to alter the appearance or form of an appliance from an orthosis that is appropriate but resisted by a patient to an orthosis that maintains its function but is embraced. This report presents the case of a patient who required a thumb orthosis to maintain her thumb interphalangeal joint in extension after a stroke but found that it interfered with her self-image and her ability to perform her job. PMID- 11887130 TI - Dealing in uncertainty. PMID- 11887131 TI - Remifentanil for use during conscious sedation in outpatient oral surgery. AB - PURPOSE: Remifentanil is a new, short-acting opioid that is similar pharmacodynamically to currently available opioids but differs in its pharmacokinetics. In the present study, we compared the use of remifentanil with the use of meperidine during intravenous conscious sedation for third molar surgery. PATIENTS AND METHODS: Forty patients who were scheduled for the removal of impacted third molars were randomly assigned to undergo 1 of 2 intravenous conscious sedation techniques. For both groups, 50:50 nitrous oxide oxygen were administered via nasal hood, and midazolam was titrated to Verril's sign. Twenty patients each then received either remifentanil 0.05 microgram/kg/min or meperidine 50 mg. Both patients and surgeons were blinded to the narcotic that was used. Blood pressure, heart rate, and oxygen saturation were determined before sedation and every 5 minutes during surgery. Recovery was measured using serial Trieger tests every 5 minutes after surgery. Patient and surgeon satisfaction of the quality of sedation was measured with a visual analog scale. RESULTS: Peak heart rate (91 beats/min for remifentanil vs 107 beats/min for meperidine, P <.01) and peak systolic blood pressure (131 mm Hg for remifentanil vs. 142 mm Hg for meperidine, P <.05) were significantly lower for the remifentanil group. Although there was a trend toward increased surgeon satisfaction with remifentanil (86 of 100 with remifentanil vs. 73 of 100 with meperidine), it was not found to be statistically significant. Likewise, other physiologic parameters were not found to be statistically significant. CONCLUSIONS: The lower peak heart rate and systolic blood pressure levels indicate that remifentanil may allow for less fluctuation in cardiovascular parameters. This could prove beneficial in patients with cardiovascular compromise. PMID- 11887133 TI - A practical method for describing patterns of tongue-base narrowing (modification of Fujita) in awake adult patients with obstructive sleep apnea. AB - PURPOSE: The purpose of the present study was to evaluate a clinically practical classification system for tongue-base narrowing and to assess intrarater and interrater reliability of the proposed system. PATIENTS AND METHODS: A population of 248 consecutive patients with polysomnographically proven upper airway obstructive pathology were evaluated (using the hypotonic method) for evidence of upper airway narrowing with the commonly available techniques of fiberoptic nasopharyngoscopy, clinical examination and lateral cephalometric analysis. RESULTS: Four basic patterns of tongue-base narrowing have been discerned to occur in awake adult patients with diagnosed obstructive upper airway pathology. These include type A (high tongue base), type B1 (high tongue base with retroepiglottic narrowing), type B2 (diffuse tongue-base narrowing), and type 3 (isolated retrogepiglottic narrowing). These recognized patterns have been found to have high intraexaminer and interexaminer reliability. No statistical correlation was found between tongue-base pattern and severity of obstructive sleep apnea, age, or facial skeletal pattern. Males had a higher percentage of type C pattern, and the type A pattern patients tended to have a higher body mass index. CONCLUSIONS: Potential future uses of this classification system include the ability to more practically assess and describe anatomic locations of tongue base narrowing and to permit an improved means of comparing the results of various surgical and nonsurgical therapies. PMID- 11887135 TI - Use of conservative condylectomy for treatment of osteochondroma of the mandibular condyle. AB - PURPOSE: The aim of the present study was to present a conservative condylectomy technique (condylectomy performed below the condylar head but high in the condylar neck) and articular disc repositioning as the surgical treatment approach for management of osteochondroma of the head of the mandibular condyle. PATIENTS AND METHODS: Six patients (4 females and 2 males) with an average age of 22.3 years (range, 13 to 32 years) and with an osteochondroma of the mandibular condyle were treated with conservative condylectomy. The remaining condylar neck stump was recontoured, and the articular disc was repositioned and stabilized over the "new" condyle. Any indicated orthognathic surgical procedures were then performed to optimize occlusion, function, and aesthetics. Clinical and radiographic evaluation was performed before surgery (T1), immediately after surgery (T2), and at the longest follow-up (T3). RESULTS: Average follow-up for the patients was 51 months (range, 22 to 108 months). No recurrence of the tumor was encountered in any of the cases. Subjective and objective evaluations of postsurgical temporomandibular joint function and range of mandibular motion were normal. Associated maxillary and/or mandibular orthognathic procedures were found to be stable in the long term. CONCLUSION: Conservative condylectomy with recontouring of the residual condylar neck to function as a condyle and repositioning of the articular disc is a viable option for treatment of osteochondromas of the mandibular condyle. The use of this method of treatment permits effective removal of the tumor and eliminates the need for autogenous grafts or total joint prostheses for temporomandibular joint reconstruction. PMID- 11887137 TI - Radiotherapy for metastases to the mandible in children. AB - PURPOSE: We present a retrospective review of all children treated since 1979 at our institution with radiotherapy for symptomatic metastases that involve the mandible. PATIENTS AND METHODS: Nine children were treated with 1 or more courses of radiotherapy for symptomatic metastases that involve the mandible. Six children had a neuroblastoma, 1 had angiosarcoma of the liver, 1 had adenocarcinoma of the rectum, and 1 had peripheral primitive neuroectodermal tumor (Ewing's sarcoma) of the spine. In 3 children, the mandible was the first bone involved by metastases. Seven children were treated with short intensive courses of radiotherapy consisting of 1 to 3 fractions to a total dose of 400 to 1,200 cGy. One child received 2,400 cGy in 6 fractions, and another child received 3,000 cGy in 10 fractions. Three children were treated with second courses of radiotherapy at 1, 2, and 5 months, respectively, from the initial course of radiotherapy. All children had received chemotherapy. RESULTS: All children died of disseminated disease at 5 to 59 months from their initial diagnosis, 5 to 29 months from the detection of metastases to bone, and only 6 days to 17 months (median, 2 months) from the first treatment of metastases to the mandible. CONCLUSIONS: The outlook for children with metastases that involve the mandible is very poor, and we recommend short intensive courses of radiotherapy consisting of 1 to 3 treatments to total doses of 400 to 1,200 cGy for palliation of pain. PMID- 11887138 TI - Titanium mesh and hydroxyapatite cement cranioplasty: a report of 20 cases. AB - PURPOSE: This article describes the use of titanium mesh and hydroxyapatite cement constructs for the treatment of large through-and-through calvarial defects. PATIENTS AND METHODS: Twenty consecutive calvarial defects (10 to 156 cm(2)) that resulted from surgical removal of neoplasms or were secondary to trauma were reviewed retrospectively after reconstruction with titanium mesh and hydroxyapatite cement. All patients were followed up by clinical examination and periodic radiographic studies for a minimum of 6 months (range, 6 months to 3 years). Three patients underwent biopsy of the construct at various points during their follow-up. RESULTS: There was no evidence of adverse healing, wound infection, or implant exposure or extrusion in any of the patients reviewed. Adequate 3-dimensional aesthetic restoration of calvarial contour was noted in each case. There was evidence of osseous ingrowth into the titanium mesh and hydroxyapatite cement construct in all 3 patients who underwent biopsy. CONCLUSION: Titanium mesh and hydroxyapatite cement cranioplasty appears to be a reasonable method for the reconstruction of significant calvarial defects. PMID- 11887139 TI - A prospective 1-year clinical and radiographic study of implants placed after maxillary sinus floor augmentation with bovine hydroxyapatite and autogenous bone. AB - PURPOSE: The purposes of this study were 1) to evaluate the survival rate of implants placed in maxillary sinuses augmented with bovine hydroxyapatite and autogenous bone 6 months before implant surgery and 2) to estimate dimensional changes of the bone graft with time using a new radiographic method. PATIENTS AND METHODS: Thirty maxillary sinuses in 20 consecutive patients with severe resorption (mean, 3.8 mm of remaining alveolar bone) were augmented with a mixture of 80% bovine hydroxyapatite and 20% autogenous bone mixed with fibrin glue to enable the placement of screw-shaped dental implants. After 6 months of primary healing, 108 implants were placed and followed with clinical and radiographic examinations during the first year of loading. Measurements of changes in height, width, and length of the grafted material were made on tomographic Scanora (Soredex Orion Corporation Ltd, Helsinki, Finland) and panoramic radiographs taken 3 and 12 months after grafting and after 1 year of bridge loading. RESULTS: Ten implants in 6 patients were lost during the study (9 before loading and 1 after 1 year of functional loading), for a survival rate of 90.7%. All patients received fixed restorations, and the bridge survival rate was 100% after 1 year of loading. Small (<10%) but statistically significant dimensional changes in the grafted material were seen during the study period. CONCLUSIONS: Acceptable short-term results can be obtained with implants placed after the use of bovine hydroxyapatite and autogenous bone for maxillary sinus floor augmentation. These grafts show good resistance to resorption. PMID- 11887141 TI - Reproducibility of maxillary positioning in Le Fort I osteotomy: a 3-dimensional evaluation. AB - PURPOSE: In the present study, we evaluated the difference between the model surgery movement and the actual surgical movement in the horizontal (X), vertical (Y), and transverse (Z) directions using the same reference coordinates. PATIENTS AND METHODS: Twelve patients (6 male and 6 female, mean age of 24.3 years) who underwent Le Fort I osteotomy and sagittal split ramus osteotomy were included in the study. The maxillary position was controlled by an intermediate splint and face-bow/bite-fork combination system. A coordinate transformation system with transition matrices was developed, which enabled objective comparison between the planned surgical change of the maxilla on the articulator and the actual surgical change assessed by the 3-dimensional cephalogram. RESULTS: The absolute mean difference was 2.2 mm. The difference between the model surgery and the surgical result ranged from -7.7 mm to 6.6 mm. The surgical result differed from the planned surgical movement by more than 2 mm in more than 45% of the measured coordinate values. CONCLUSION: Although all of the patients were satisfied with their postsurgical appearance and occlusion, the result shows that further development is required in the maxillary positioning system. PMID- 11887142 TI - AO self-drilling and self-tapping screws in rat calvarial bone: an ultrastructural study of the implant interface. AB - PURPOSE: The purpose of the present study was to compare the bone-to-screw interface of both AO self-drilling screws (SDSs) and self-tapping screws (STSs) using scanning electron microscopy. MATERIALS AND METHODS: The calvaria from Sprague-Dawley rats were harvested, and the periosteum was removed. The samples were stored in saline until use. AO STSs and SDSs were then inserted into the outer surface of the calvaria without irrigation. All screws were 4 mm in length and were inserted until their tips protruded to the endosteal side. A total of 6 screws, 3 of each type, were placed. The dimensions of the screws were 1.3, 1.5, and 2.0 mm. All STSs were placed with an appropriately sized predrilled hole. Samples were then examined and digitally photographed using scanning electron microscopy. RESULTS: No damage was noted to any of the implants (STSs or SDSs). Excellent adaptation of the STSs to host bone was observed for all specimens. On the other hand, the endosteal surface of the SDSs demonstrated large voids adjacent to the screw threads at the interface. These appeared to represent microfractures of the bone at the bone-implant interface. CONCLUSIONS: These data suggest a greater amount of bone damage during placement of the SDSs compared with the STSs. These results point to the need for further investigation into the use of SDSs in clinical practice. PMID- 11887145 TI - Distraction osteogenesis of the mandible in the previously irradiated patient. PMID- 11887144 TI - A firm, bluish mass of the cheek in a 17-month-old child. PMID- 11887146 TI - Malignant hemangiopericytoma of the infratemporal fossa: report of a case. PMID- 11887147 TI - Unusual congenital midline cervical anomaly. PMID- 11887148 TI - Bilateral discal tumoral calcinosis of the temporomandibular joint. PMID- 11887149 TI - Adenocarcinoma of the lacrimal gland: report of a case. PMID- 11887150 TI - Bilateral necrosis of the tongue consecutive to cardiac arrest. PMID- 11887151 TI - Surgical (implantation) cyst of the mandible with ciliated respiratory epithelial lining: a case report. PMID- 11887152 TI - Hyperactivity-induced suprahyoid muscular hypertrophy secondary to excessive video game play: a case report. PMID- 11887153 TI - Rhinocerebral mucormycosis in a patient with latent diabetes mellitus: a case report. PMID- 11887154 TI - Skeletal muscle autograft for repair of the human inferior alveolar nerve: a case report. PMID- 11887155 TI - Sewing needle in the maxillary antrum. PMID- 11887156 TI - The use of a polyurethane skull replica as a template for contouring titanium mesh. PMID- 11887157 TI - Third molars and eating disorders. PMID- 11887158 TI - Another view on the anticoagulated patient. PMID- 11887159 TI - The effects of phospholipase C inhibition on insulin-stimulated glucose transport in skeletal muscle. AB - Previous research has demonstrated that phospholipase C (PLC) is involved in insulin-stimulated glucose transport in 3T3-L1 adipocytes. The purpose of the current investigation was to determine if PLC is also involved in insulin stimulated glucose uptake in rat skeletal muscle. To that end, we used an in vitro muscle preparation of the rat soleus muscle to test the effects of the PLC inhibitor, U73122, on glucose transport. The PLC inhibitor, U73122, led to a concentration-dependent inhibition of insulin (0.6 nmol/L)-stimulated glucose transport, whereas it had no effect on basal glucose transport. Specifically 10, 20, 50, and 150 micromol/L U73122 inhibited insulin (0.6 nmol/L)-stimulated glucose transport approximately 17%, 20%, 26%, and 38%, respectively, while an equal molar concentration of U73343 (inactive form of U73122) and/or carrier media (dimethyl sulfoxide [DMSO]) did not influence glucose uptake. A secondary aim of this investigation was to determine if increasing the concentration of insulin from a physiologic concentration (0.6 nmol/L) to a supraphysiologic concentration (6.0 nmol/L) could ameliorate the inhibitory effects of U73122. A 10-fold increase in insulin eliminated the inhibitory effects of U73122 on insulin-stimulated glucose uptake in soleus muscle. In summary, this preliminary report provides evidence to suggest that a PLC signaling mechanism modifies insulin-stimulated glucose uptake in skeletal muscle via its influence on insulin sensitivity. PMID- 11887160 TI - Impact of intraportal N(omega)-nitro-L-arginine infusion on hepatic glucose metabolism in total parenteral nutrition-adapted dogs: interaction with infection. AB - During chronic total parenteral nutrition (TPN), liver glucose uptake and lactate release are markedly elevated. However, in the presence of an infection, hepatic glucose uptake and lactate release are reduced. Glucose delivery (the product of liver blood flow and inflowing glucose concentration) is a major determinant of liver glucose uptake. Hepatic blood flow is increased during infection, and increased nitric oxide (NO) biosynthesis is thought to contribute to the increase. Our aim was to determine if the increase in liver blood flow served to limit the infection-induced decrease in hepatic glucose uptake and metabolism. Chronically catheterized conscious dogs received TPN for 5 days at a rate designed to match daily basal energy requirements. On the third day of TPN administration, a sterile (SHAM) or Escherichia coli (E. coli)-containing (INF) fibrin clot was implanted in the peritoneal cavity. Forty-two hours later, somatostatin was infused with intraportal replacement of insulin (10 +/- 2 v 23 +/- 2 microU/mL, SHAM v INF, respectively) and glucagon (22 +/- 4 v 90 +/- 8 pg/mL) to match concentrations observed in sham and infected animals. Tracer and arteriovenous difference techniques were used to assess hepatic glucose metabolism. Following a 120-minute basal sampling period, sham and infected animals received either intraportal saline or N(omega)-nitro-L-arginine (L-NNA; 37 microg x kg(-1) x min(-1)) infusion for 180 minutes. Isoglycemia (120 mg/dL) was maintained with a variable glucose infusion. In the infected group L-NNA infusion decreased hepatic arterial blood flow (23.3 +/- 0.7 to 8.6 +/- 0.5 mL x kg(-1) x min(-1)), but not portal vein blood flow. Neither portal vein nor hepatic artery blood flow were altered by L-NNA infusion in the sham group. Hepatic glucose uptake and lactate metabolism were not altered by L-NNA infusion in either group. In summary, during infection, an increase in NO biosynthesis contributes to the increase in hepatic arterial blood flow, while it exerts no effect on hepatic glucose metabolism. PMID- 11887161 TI - Genetic and environmental regulation of Na/K adenosine triphosphatase activity in diabetic patients. AB - Even if the pathogenesis of diabetic neuropathy is incompletely understood, an impaired Na/K adenosine triphosphatase (ATPase) activity has been involved in this pathogenesis. We previously showed that a restriction fragment length polymorphism (RFLP) of the ATP1-A1 gene encoding for the Na/K ATPase's alpha 1 isoform is associated with a low Na/K ATPase activity in the red blood cells (RBCs) of type 1 diabetic patients. We thus suggested that the presence of the variant of the ATP1A1 gene is a predisposing factor for diabetic neuropathy, with a 6.5% relative risk. Furthermore, there is experimental evidence showing that lack of C-peptide impairs Na/K ATPase activity, and that this activity is positively correlated with C-peptide level. The aim of this study was to evaluate the respective influence of genetic (ATP1-A1 polymorphism) and environmental (lack of C-peptide) factors on RBC's Na/K ATPase activity. Healthy and diabetic European and North African subjects were studied. North Africans were studied because there is a high prevalence and severity of neuropathy in this diabetic population, and ethnic differences in RBC's Na/K ATPase activity are described. In Europeans, Na/K ATPase activity was significantly lower in type 1 (285 +/- 8 nmol Pi/mg protein/h) than in type 2 diabetic patients (335 +/- 13 nmol Pi/mg protein/h) or healthy subjects (395 +/- 9 nmol Pi/mg protein/h). Among type 2 diabetic patients, there was a significant correlation between RBC's Na/K ATPase activity and fasting plasma C-peptide level (r = 0.32, P <.05). In North Africans, we confirm the ethnic RBC's Na/K ATPase activity decrease in healthy subjects (296 +/- 26 v 395 +/- 9 nmol Pi/mg protein/h, r < 0.05), as well as in type 1 diabetic patients (246 +/- 20 v 285 +/- 8 nmol Pi/mg protein/h; P <.05). However, there is no relationship between the ATP1A1 gene polymorphism and Na/K ATPase activity. ATP1A1 gene polymorphism could not explain the ethnic difference. We previously showed that Na/K ATPase activity is higher in type 1 diabetic patients without the restriction site on ATP1A1 than in those heterozygous for the restriction site. This fact was not observed in healthy subjects. In type 2 diabetic patients, association between ATP1A1 gene polymorphism and decreased enzyme activity was found only in patients with a low C-peptide level. Therefore, the ATP1-A1 gene polymorphism influences Na/K ATPase activity only in case of complete or partial C-peptide deficiency, as observed in type 1 and some type 2 diabetic patients, without any correlation with hemoglobin A1c (HbA1c). Correlation observed between C-peptide levels and RBC's Na/K ATPase suggests that the deleterious effect of C peptide deficiency on Na/K ATPase activity is worse in the presence of the restriction site. This may explain the high relative risk of developing the neuropathy observed in type 1 diabetic patients bearing the variant allele. PMID- 11887162 TI - Serum leptin concentrations in children with type 1 diabetes mellitus: relationship to body mass index, insulin dose, and glycemic control. AB - Although obesity is a frequent feature of type 2 diabetes mellitus (DM), many patients with type 1 DM are prone to high body mass index (BMI). We measured serum leptin concentrations in a cohort of children (n = 55) with type 1 diabetes mellitus (DM), as well as their anthropometric parameters including BMI, skin fold thickness at multiple sites, and midarm circumference. Glycemic control was assessed by blood glucose (BG) monitoring before meals, and measurement of glycated hemoglobin (HbA1c) and insulin dose/kg/d was recorded. Dietary evaluation and assessment of caloric intake (kg/d) was performed by an expert dietitian. In the newly diagnosed children (n = 10) before initiation of insulin therapy, circulating leptin concentration was significantly lower (1.1 +/- 0.8 ng/dL) versus 5 days after insulin therapy (1.45 +/- 0.7 ng/dL). The decreased leptin level appears to be related to insulinopenia in these patients. In 45 children with type 1 DM on conventional therapy (2 doses of insulin mixture (NPH and regular) subcutaneous (SC) before breakfast and dinner for more than 2 years), serum leptin concentration was significantly higher (2.15 +/- 1 ng/dL) compared with age-matched normal children (1.3 +/- 1 ng/dL). Diabetic children were further divided into 2 groups according to their HbA1c level: group 1 with HbA1C less than 7.5% (less than 2 SD above the mean for normal population) (n = 29) and group 2 with HbA1c greater than 7.5%. (greater than 2 SD above the mean for normal population) (n = 16). Patients with a higher HbA1c level (group 2) had a higher leptin concentration (2.3 +/- 0.8 ng/dL), higher BMI (17.8 +/- 1.7), and were receiving higher insulin dose/kg (0.92 +/- 0.2 U/kg/d) compared with group 1 (lower HbA1c) (1.78 +/- 0.8 ng/dL, 16.7 +/- 1.5, and 0.59 +/- 0.2 U/kg/d, respectively). Group 2 patients had a higher incidence of late morning hypoglycemia (9/29) versus group 1 patients (2/16). Analysis of dietary intake showed that patients with a higher HbA1c (group 2) consumed more calories (73.5 +/- 10.5 kcal/kg/d) versus patients with lower HbA1c (64.2 +/- 8.7 kcal/kg/d). These findings pointed to the unphysiologic nature of injecting a mixture of insulin twice daily. To cover the relatively big lunch meal (40% to 50% of the total caloric intake in the Arab countries) and prevent afternoon hyperglycemia, there is a great tendency to increase NPH dose before breakfast. This, in turn, induces late-morning hypoglycemia and increases appetite and food intake at that time. Multiple regression analysis showed that circulating leptin concentrations (the dependent variable) were best correlated with the mean skinfold thickness (SFT), BMI, and caloric intake/kg/d (together they explained 65% of the variability in leptin concentrations). It appears that oversubstitution by insulin and increased food intake stimulate fat synthesis and subsequently BMI. Increased appetite and BMI contribute to increased leptin secretion and explains the higher leptin levels in undercontrolled diabetic children (higher circulating HbA1c concentrations) who were oversubstituted by insulin. PMID- 11887163 TI - Interaction between glucose metabolism and endogenous insulin release in hypertension. AB - The minimal model approach was applied to examine the dynamic interaction between glucose metabolism and endogenous insulin release during an intravenous glucose tolerance test (IVGTT) in a group of hypertensive patients (H group) compared with a group of normotensive subjects (N group). A modified version of the classical minimal model of C-peptide kinetics and secretion was used to evaluate the total amount of insulin secretion per unit of distribution volume (TIS) together with 3 indexes of beta-cell function (the basal, Phi(b), first, Phi1, and second phase, Phi2, beta-cell sensitivity to glucose). These indexes were associated with estimates of glucose effectiveness (S(G)) and insulin sensitivity (S(I)) provided by the classical minimal model of glucose kinetics. No significant differences were found in Phi(b), Phi1, and Phi2 estimates between the H group and the N group. In the H group, the average TIS was 54% higher (P <.05) than in the N group, while S(G) and S(I) estimates showed a 44% decrease (P <.05) and a 51% decrease (P <.05), respectively. These results suggest that hyperglycemia observed in our H group during IVGTT is a compensatory response to insulin resistance (low S(I)) and to the reduced ability of glucose to promote its own metabolism (low S(G)). This hyperglycemic state causes a larger than normal stimulation of beta cell, which explains insulin hypersecretion (higher TIS) even in the presence of normal beta-cell sensitivity values of Phi(b), Phi1, and Phi2. PMID- 11887164 TI - Reduced leptin concentrations in subjects with type 2 diabetes mellitus in Sudan. AB - Differences have been observed in the relationship between leptin and metabolic perturbations in glucose homeostasis. Because no information is available from indigenous African populations with diabetes, the purpose of this study was to investigate the possible associations between leptin and different clinical and biochemical characteristics of a large group of subjects with type 2 diabetes mellitus in Sudan. A total of 104 (45 men and 59 women) consecutive type 2 diabetes patients and 75 control subjects (34 men and 41 women) were studied. The body mass index (BMI), blood glucose, serum insulin, and proinsulin were measured and related to serum leptin concentrations. Leptin was higher in females than in males and correlated significantly to BMI. The main novel finding was that serum leptin was significantly lower in diabetic subjects compared with controls in both females (P =.0001) and males (P =.019), although BMI did not differ between diabetic and nondiabetic subjects. Diabetic subjects treated with sulphonylurea (n = 81) had lower BMI than those treated with diet alone or other hypoglycemic drugs (n = 23) (P =.0017), but there was no difference in leptin levels between the 2 groups after adjustment for BMI (P =.87). In diabetic subjects, serum leptin correlated positively with the homeostatic assessment (HOMA) of both beta cell function (P =.018) and insulin resistance (P =.038), whereas in control subjects, leptin correlated with insulin resistance (P =.0016), but not with beta cell function. Diabetic subjects had higher proinsulin levels (P =.0031) and higher proinsulin to insulin ratio (P =.0003) than nondiabetic subjects. In univariate analysis, proinsulin showed a weak correlation to leptin (P =.049). In conclusion, we show in a large cohort of Sudanese subjects with type 2 diabetes that circulating leptin levels are lower in diabetic subjects than in controls of similar age and BMI. The lower serum leptin in diabetic subjects may be a consequence of differences in fat distribution. PMID- 11887165 TI - Metabolic response to a C-glucose load in human immunodeficiency virus patients before and after antiprotease therapy. AB - Changes in glucose and fat metabolism associated with human immunodeficiency virus (HIV) infection have received attention because of the development of glucose intolerance, dyslipidemia, and lipodystrophy associated with protease inhibitor (PI) therapy. The response to ingested [13C]glucose (1.4 g/kg) was determined in 9 asymptomatic male HIV patients before and after 4.8 months of PI therapy (nelfinavir, 2,250 mg/d) compared with 9 matched seronegative HIV controls. No significant difference was observed for basal plasma glucose, insulin, and C-peptide concentrations between controls and patients before PI therapy. After 4.8 months of PI therapy, basal plasma glucose concentration was slightly, but significantly, increased (approximately 15%) compared with controls or HIV patients prior to receiving PI therapy. Over the first hour following ingestion of the glucose load, plasma glucose and insulin concentrations were higher in HIV patients than in controls, both before (approximately 15% and approximately 29%, respectively) and after (approximately 32% and approximately 43%, respectively) PI therapy. In addition, plasma C-peptide concentration was approximately 61% higher after PI therapy. The oxidation rate of fat, endogenous, and exogenous glucose was computed from the VO2 and respiratory exchange ratio corrected for protein oxidation and from 13C/12C in expired CO2. The only difference between controls and patients both before and after PI therapy was observed over the first 120 minutes following ingestion of the glucose load, when HIV patients oxidized approximately 18% more glucose and approximately 19% less fat than controls. This was not due to a larger oxidation rate of exogenous glucose, but to a larger oxidation rate of endogenous glucose (approximately 50%) in patients compared with controls. These data indicate that HIV infection is associated with minor changes in glucose metabolism, and that PI therapy with nelfinavir for 4.8 months only slightly further impairs glucose metabolism as assessed in response to a large oral glucose load. However, the larger stimulation of total and endogenous glucose oxidation and the larger reduction in fat oxidation, observed in the metabolic response to the glucose load in HIV patients, over time, could result in the accumulation of body fat and could contribute to lipodystrophy. PMID- 11887167 TI - Histochemical evidence for pathways insulin cells use to oxidize glycolysis derived NADH. AB - The activity of lactate dehydrogenase is known to be low in the pancreatic beta cell, and the activity of the mitochondrial glycerol phosphate dehydrogenase (mGPD), the key enzyme of the glycerol phosphate shuttle, is known to be high in this cell. Lactate dehydrogenase was demonstrated histochemically in insulin positive cells of the rat pancreas, and its activity was semiquantified densitometrically; activity in these cells was estimated to be about 8% of that in the surrounding acinar tissue. mGPD histochemical activity was extremely high in cells exhibiting insulin immunofluorescence, while activity in surrounding pancreas tissue was negligible. When the activity was measured in situ at a physiologic concentration of substrate, this enzyme was inactive in the absence of free calcium. These results are consistent with the idea that glucose, the most potent physiologic insulin secretagogue, stimulates insulin secretion via aerobic glycolysis. If glycolysis-derived NADH, instead of being reoxidized in a mitochondrial hydrogen shuttle, is reoxidized to NAD by pyruvate via the reaction catalyzed by lactate dehydrogenase with the resulting formation of lactate, there would be little or no pyruvate available for mitochondrial metabolism. Consequently adenosine triphosphate formation would be about 5% to 7% of that formed by the complete combustion of glucose to carbon dioxide via mitochondrial metabolism. The low lactate dehydrogenase and high mGPD emphasize the importance of mitochondrial hydrogen shuttles for reoxidation of glycolysis-derived NADH in insulin secretion. PMID- 11887166 TI - Effects of pioglitazone on metabolic parameters, body fat distribution, and serum adiponectin levels in Japanese male patients with type 2 diabetes. AB - The aim of this study was to evaluate the effects of pioglitazone on clinical and metabolic parameters, body fat distribution, and serum adiponectin, a recently discovered antiatherosclerotic hormone, in Japanese patients with type 2 diabetes. Ten male patients aged 40 to 66 (57.7 +/- 7.4) years, who were being treated with dietary therapy alone (n = 7) or with a stable dose of sulfonylurea (n = 3), were studied at baseline and after 3 months of pioglitazone treatment (30 mg/d). Body mass index (BMI), blood pressure, fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), serum insulin, adiponectin, and lipid profile were measured. Also, visceral adipose tissue area (VAT) and subcutaneous adipose tissue area (SAT) at the umbilical level were determined by computed tomographic (CT) scanning. Mean blood pressure (109 +/- 14 to 101 +/- 10 mm Hg), FPG (8.6 +/- 1.4 to 7.0 +/- 1.0 mmol/L), serum insulin (54 +/- 11 to 30 +/- 8 pmol/L, P <.01 for all), and HbA1c (6.7 +/- 0.8 to 6.1% +/- 0.6%, P =.013) decreased significantly during 3 months of pioglitazone treatment. BMI (26.4 +/- 3.2 to 27.0 +/- 3.5 kg/m2), low-density lipoprotein (LDL) cholesterol (124 +/- 24 to 138 +/- 24 mg/dL) and SAT (155 +/- 69 to 179 +/- 81cm2, P <.05 for all) increased, while triglycerides and high-density lipoprotein (HDL) cholesterol did not change significantly after 3 months of pioglitazone treatment. Serum adiponectin level increased in all patients (4.8 +/- 1.7 to 14.4 +/- 2.1 microg/mL, P =.005). VAT tended to increase (165 +/- 38 to 180 +/- 46 cm2) and VAT/SAT ratio tended to decrease (1.2 +/- 0.3 to 1.1 +/- 0.3), but these differences did not reach statistical significance. These results suggest that pioglitazone exerts good glycemic and blood pressure control despite increased BMI and SAT in Japanese male patients with type 2 diabetes. It is also suggested that pioglitazone may have an antiatherosclerotic effect by increasing serum adiponectin level. PMID- 11887168 TI - Effect of menopausal status on lipolysis: comparison of plasma glycerol levels in middle-aged, premenopausal and early, postmenopausal women. AB - To determine whether menopausal status affects systemic lipolysis, we measured plasma glycerol concentrations following an overnight fast and during euglycemic hyperinsulinemic conditions (40 mU x m(-2) x min(-1)) in 43 middle-aged, premenopausal women (mean +/- SE; 47 +/- 0.4 years) and 26 early, postmenopausal (51 +/- 0.8 years) women. In addition, body composition was measured by dual energy x-ray absorptiometry and abdominal fat distribution by computed tomography (CT). Postmenopausal women had greater amounts of whole body (fat mass, 22.8 +/- 1.4 v 17.4 +/- 1.2 kg; percent fat, 34.7 +/- 1.2 v 29.1 +/- 1.4; both P <.01) and intra-abdominal fat (89.0 +/- 6.5 v 55.9 +/- 4.4 cm2; P <.01) compared with premenopausal women. Despite greater adiposity, plasma glycerol concentrations were similar between pre- and postmenopausal women following an overnight fast (142.7 +/- 9.7 v 136.1 +/- 6.4 micromol/L) and at 30 minutes (112.7 +/- 5.5 v 108.4 +/- 4.5 micromol/L ) and 120 minutes (92.7 +/- 4.5 v 97.5 +/- 5.9 micromol/L ) into the euglycemic hyperinsulinemic clamp. Plasma glycerol levels remained similar after statistical adjustment for fat mass, percent fat, and intra-abdominal fat. Moreover, no differences in plasma glycerol were observed in pre- and postmenopausal women matched (+/- 5%) for fat mass (n = 22/group) or intra-abdominal fat (n = 15/group). In premenopausal women, plasma glycerol levels at 30 and 120 minutes of hyperinsulinemia were positively related to adiposity measures (range, r =.314 to r =.493; P <.05 to P <.01), although no relationships were found in postmenopausal women. Our results suggest no effect of menopausal status on plasma glycerol levels under postabsorptive or hyperinsulinemic conditions. PMID- 11887169 TI - Linoleic acid-stimulated vascular adhesion molecule-1 expression in endothelial cells depends on nuclear factor-kappaB activation. AB - Endothelial activation is an important step in atherogenesis. In addition to established cardiovascular risk factors, such as hypercholesterolemia, hypertension, diabetes mellitus, and homocysteinemia, high plasma levels of triglyceride-rich lipoproteins may be an important cause of endothelial activation as well. Free fatty acids hydrolyzed from core triglycerides of these particles can exert both pro- and anti-inflammatory effects on the vascular wall. omega-3 fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), have been shown to inhibit cytokine-induced endothelial activation. In contrast, we and others have previously shown that the omega-6 fatty acid linoleate activates transcription factor nuclear factor-kappaB (NF-kappaB) in endothelial cells. In this study, we show that linoleic acid stimulates vascular adhesion molecule-1 (VCAM-1) protein and mRNA expression in cultured human endothelial cells, as assessed by immunofluorescence and Northern blotting. Release of shedded soluble VCAM-1 from cultured human endothelial cells was also increased by the addition of linoleic acid, as determined by enzyme-linked immunosorbent assay (ELISA). By use of cultured rat aortic endothelial cells transfected with an IkappaB super-repressor (DeltaN2 cells), we provide evidence that NF-kappaB signalling is required in the linoleic acid-induced VCAM-1 expression in endothelial cells, whereas other transcription factors appear to be involved in the increased endothelial plasminogen activator inhibitor-1 (PAI-1) production in response to linoleic acid. These findings suggest that diets rich in linoleic acid may be proinflammatory and thus atherogenic by activating vascular endothelial cells. PMID- 11887170 TI - Atorvastatin treatment beneficially alters the lipoprotein profile and increases low-density lipoprotein particle diameter in patients with combined dyslipidemia and impaired fasting glucose/type 2 diabetes. AB - Diabetic dyslipidemia is featured by hypertriglyceridemia, low high-density lipoprotein (HDL) cholesterol levels, and elevated low-density lipoprotein (LDL) cholesterol commonly in the form of small, dense LDL particles. First-line treatment, fibrates versus statins or both, of dyslipidemia in diabetic patients has been the focus of debate. We investigated the potential hypolipidemic effects of atorvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor with good triglyceride lowering properties, in patients with combined dyslipidemia and evidence of impaired fasting glucose or type 2 diabetes. Twenty patients were recruited for the study, and after a 60-day wash out period, baseline measurements of lipoprotein parameters, LDL particle diameter, and apolipoprotein B (apoB) degradation fragments were obtained. The group was then randomized, in a double-blinded manner, into 2 subgroups. Group A received atorvastatin (80 mg) and group B received placebo daily for 60 days. After the first treatment period, all patients were reanalyzed for the above parameters. The treatment regime then crossed over for the second treatment period in which group A received placebo and group B received atorvastatin (80 mg) daily for 60 days. All parameters were remeasured at the end of the study. Treatment with atorvastatin resulted in a statistically significant reduction in total cholesterol (41%), LDL cholesterol (55%), triglycerides (TG) (32%), and apoB (40%). Mean LDL particle diameter significantly increased from 25.29 +/- 0.24 nm (small, dense LDL subclass) to 26.51 < 0.18 nm (intermediate LDL subclass) after treatment with atorvastatin (n = 20, P <.005). At baseline, LDL particles were predominantly found in the small, dense subclass; atorvastatin treatment resulted in a shift in the profile to the larger and more buoyant LDL subclass. Atorvastatin treatment did not produce consistent changes in the appearance of apoB degradation fragments in plasma. Our results suggest that atorvastatin beneficially alters the atherogenic lipid profile in these patients and significantly decreases the density of LDL particles produced resulting in a shift from small, dense LDL to more buoyant and less atherogenic particles. PMID- 11887171 TI - Relative importance of sympathetic outflow and insulin in the reactivation of brown adipose tissue lipogenesis in rats adapted to a high-protein diet. AB - The effect of denervation or acute insulin deficiency on brown adipose tissue lipogenesis was investigated in rats adapted to a high-protein diet before and after diet reversion to a balanced diet. Denervation of rats fed the balanced diet induced a 50% reduction in in vivo rates of brown adipose tissue fatty acid synthesis, with decreased activities of acetyl-CoA carboxylase and adenosine triphosphate (ATP)-citrate lyase. The markedly (80%) reduced fatty acid synthesis and enzyme activities in brown adipose tissue from rats adapted to the high protein diet were not affected by denervation. Replacement of the high-protein diet by the balanced diet for 24 hours restored fatty acid synthesis to normal levels, but recovery of enzyme activities was only partial. Lipogenesis restoration and partial recovery of enzyme activities were impaired in denervated tissue from high-protein diet-fed rats. In all experimental conditions, the activities of acetyl-CoA carboxylase and ATP-citrate lyase showed a better correlation with brown adipose tissue lipogenesis than the generators of H(+), glucose-6-P dehydrogenase, and malic enzyme. Anti-insulin serum administration during the 12- to 24-hour period after diet reversion completely blocked lipogenesis recovery in innervated and denervated tissues and drastically reduced brown adipose tissue lipogenesis of concomitantly injected rats fed the balanced diet. The data suggest that efficient and rapid adjustments of brown adipose tissue lipogenesis require sympathetic activation, and that this tissue can maintain significant, albeit reduced, rates of lipogenesis in the absence of sympathetic activation, but not in the absence of insulin. PMID- 11887172 TI - Unique cases of unilateral hyperaldosteronemia due to multiple adrenocortical micronodules, which can only be detected by selective adrenal venous sampling. AB - Primary aldosteronism is classified as aldosterone-producing adenoma (APA), idiopathic hyperaldosteronism (IHA), unilateral adrenal hyperplasia (UAH), primary adrenal hyperplasia (PAH), adrenal cancer, and glucocorticoid-remediable aldosteronism. We describe here 4 cases of primary aldosteronism due to unilateral hyperaldosteronemia, demonstrating unique histopathologic findings, such as unilateral multiple adrenocortical micronodules in the affected adrenals. Thirty-three patients with primary aldosteronism were consecutively admitted; 27 of them were treated by unilateral adrenalectomy. Four of them also had unilateral adrenal hypersecretion of aldosterone by selective adrenal venous sampling and adrenocortical multiple micronodules without an adenoma. These patients had hyporeninemic hyperaldosteronism with normokalemic hypertension. In these patients, furosemide plus upright test failed to increase plasma renin activity (PRA); the ratio of plasma aldosterone concentration (PAC) to PRA at 90 minutes after captopril administration was similar to that in patients with IHA and APA. Aldosterone concentrations were increased in each unilateral adrenal vein, and poorly encapsulated multiple adrenocortical micronodules from 2 to 3 mm in diameter were microscopically detected in the resected adrenal glands. Immunohistochemical analysis of steroidogenic enzymes, including cholesterol side chain cleavage, 3beta-hydroxysteroid dehydrogenase, 21-hydroxylase, 17alpha hydroxylase, and 11beta-hydroxylase, indicated that the cortical cells within these micronodules were active in aldosterone production, while the non-nodular zona glomerulosa cells were inactive. We conclude that the clinical and pathologic characteristics of our 4 cases with unilateral multiple adrenocortical micronodules (UMN) are distinct from those of APA, IHA, UAH, and PAH. Furthermore, unilateral hyperaldosteronemia induced by UMN may be frequently misdiagnosed, because standard imaging tests, which cannot always detect tiny abnormalities of adrenals, showed "normal adrenal glands" in these patients. Thus, primary aldosteronism due to UMN should be carefully examined for differential diagnosis of each form of hyperaldosteronemia. PMID- 11887173 TI - Association between vitamin D receptor polymorphism and type 2 diabetes or metabolic syndrome in community-dwelling older adults: the Rancho Bernardo Study. AB - Vitamin D receptor (VDR) polymorphism influences susceptibility to type 1 diabetes, but the association with type 2 diabetes is not clear. We investigated the association between VDR polymorphism and type 2 diabetes and metabolic syndrome in a community-based study of unrelated older adults without known diabetes. Oral glucose tolerance test (75 g), plasma glucose and insulin measurement, homeostasis model assessment (HOMA), and VDR genotyping were performed. The distributions of genotype frequencies of ApaI, BsmI, and TaqI polymorphism did not differ between persons with and without diabetes, but the frequency of aa genotype of ApaI polymorphism was marginally higher in persons with type 2 diabetes (P =.058). Fasting plasma glucose (P <.05) and prevalence of glucose intolerance (P <.05) were significantly higher in nondiabetic persons with aa genotype compared with those with AA genotype. The bb genotype of BsmI polymorphism was associated with insulin resistance as assessed by HOMA after adjustment for age, sex, body mass index (BMI), and calcium and vitamin D use in persons without diabetes (P <.05). Our research suggests that ApaI polymorphism may be associated with glucose intolerance independent of defective insulin secretion and BsmI polymorphism with insulin resistance in a nondiabetic Caucasian population. PMID- 11887174 TI - A genetic study of cortisol measured before and after endurance training: the HERITAGE Family Study. AB - The aim of this study was to investigate whether there are familial influences on cortisol levels at baseline and in response to endurance exercise training and, if so, whether there is evidence for a major gene effect. There were 476 white individuals in 99 nuclear families and 247 black individuals in 105 families with valid cortisol data in the HERITAGE Family Study. Data adjustments were carried out separately in each of 8 sex by generation by race groups, using stepwise multiple regression procedures. The familial factors underling the variability in baseline cortisol (log-transformed and adjusted for age and baseline body mass index [BMI]) and its training response (post-training minus baseline, adjusted for age, baseline BMI, and the baseline cortisol value) were assessed by estimating familial correlations and carrying out segregation analysis. In the white sample, significant familial resemblance was detected for both baseline cortisol and the training response, with maximal heritabilities of 38% and 32%, respectively. However, significant familial correlations were not detected for either cortisol phenotype in the black sample, perhaps owing, in part, to the much smaller family sizes. Results of segregation analysis of the white sample provided evidence for Mendelian additive genes influencing baseline cortisol and its training response. The major genes accounted for 33% and 31% of the variance for baseline cortisol and the training response with 48% and 5% of the sample homozygous for the genotype leading to high values, respectively. In conclusion, we found significant familial effects influencing levels of baseline cortisol and its training response in the white sample. The putative major gene effects appear to explain most of the observed familial resemblance, this will motivate further linkage and association studies. PMID- 11887175 TI - Acylcarnitine profiles in fibroblasts from patients with respiratory chain defects can resemble those from patients with mitochondrial fatty acid beta oxidation disorders. AB - Mitochondrial fatty acid beta-oxidation (FAO) is coupled to the respiratory chain (RC). Functional defects of one pathway may lead to secondary alteration in flux through the other. We investigated the acylcarnitine profiles in cultured fibroblasts obtained from 14 healthy subjects, 31 patients with 8 different primary enzyme deficiencies of FAO, and 16 patients with primary RC defects including both isolated and multiple enzyme complex defects. Intact cells were incubated in media containing deuterium-labeled hexadecanoic acid and L carnitine, and the acylcarnitines analysed using an electrospray tandem mass spectrometer. All FAO-deficient cell lines revealed disease-specific acylcarnitine profiles related to the sites of defects. Some cell lines from patients with RC defects showed profiles similar to those of controls, whereas others had abnormal profiles mimicking those found in FAO disorders. The acylcarnitine profiles of patients with RC enzyme defects were not predictable, and in some patients defects caused by mutations in either nuclear-encoded or mitochondrial DNA were associated with acylcarnitine abnormalities. While in vitro acylcarnitine profiling is useful for the diagnosis of FAO deficiencies, abnormal profiles do not exclusively indicate these disorders, and primary defects of the RC remain a possibility. Awareness of this diagnostic pitfall will aid in the selection of subsequent confirmatory tests and therapeutic options. PMID- 11887176 TI - Uric acid concentration in subjects at risk of type 2 diabetes mellitus: relationship to components of the metabolic syndrome. AB - High uric acid concentration is a common finding in subjects with risk factors for cardiovascular disease (CVD), including some characteristics of the metabolic syndrome. However, its exact role in this setting and in the progression to type 2 diabetes mellitus (DM) is not well understood and could be affected by confounding factors such as hypertriglyceridemia. Our study aimed to establish the relationship between uric acid (avoiding the interference of high triglyceride levels), insulin sensitivity, and components of the metabolic syndrome in a group of subjects at high risk of developing DM. Among 201 subjects included in the study, 111 (55.2%) showed an abnormal oral glucose tolerance and uric acid levels higher than those measured in subjects with normal glucose tolerance. Body mass index (BMI), triglycerides, diastolic blood pressure (DBP), and 2-hour glycemia in the oral glucose tolerance test (OGTT) contributed independently to uric acid concentration (R2 =.59). However, uric acid did not affect either insulin sensitivity or glucose tolerance. The recovery tests revealed that a triglyceride concentration > or = 3 mmol/L interfered with the measurement of uric acid level when a colorimetric method was used, but not when a dry-chemistry method was used. In conclusion, uric acid concentration is higher in subjects at high risk of DM with abnormal glucose tolerance and is independently determined by various components of the metabolic syndrome. PMID- 11887177 TI - Effects of dehydroepiandrosterone on rat apolipoprotein AI gene expression in the human hepatoma cell line, HepG2. AB - Serum apolipoprotein AI (apoAI) levels correlate with the risk of developing atherosclerosis. Previous studies have suggested that dehydroepiandrosterone (DHEA) lowers high-density lipoprotein (HDL)-cholesterol levels. We investigated whether or not DHEA may lower HDL-cholesterol levels by suppressing apoAI gene transcription in hepatocytes. ApoAI mRNA levels, assessed by Northern blotting, were suppressed in HepG2 cells treated with DHEA (34%) (10 microg/mL) or testosterone (36%) (T, 1 microg/mL). Estradiol alone (E2, 1 microg/mL) had relatively little effect on apoAI mRNA levels, while E2 in combination with DHEA prevented a decrease in apoAI mRNA levels compared to DHEA alone. To determine whether these effects were due to changes in apoAI gene transcription, HepG2 cells were transfected with a plasmid carrying the full-length promoter of the rat apoAI gene ligated into a chloramphenicol acetyltransferase (CAT) reporter construct. The plasmid pCMV.SPORT-beta-gal was included in each transfection to normalize the data to transfection efficiency. Cells were then cultured in the presence or absence of DHEA (10 microg/mL), T (1 microg/mL), 17alpha methyltestosterone (MTT, 1 microg/mL), 5alpha-dihydrotestosterone (DHT, 1 microg/mL), E2 (1 microg/mL), or a combination of DHEA plus E2, T plus E2, MTT plus E2, and DHT plus E2, for 24 hours. CAT activity, relative to beta galactosidase activity, was reduced by 19.6%, 57.6%, 38.6%, and 54.6% with DHEA, T, DHT, and MTT addition, respectively. E2 increased CAT activity by 43.8%. When the androgens (ie, DHEA, T, DHT, or MTT) were combined with E2, apoAI promoter activity was suppressed. We conclude, therefore, that androgens downregulate apoAI promoter activity in the presence or absence of E2. However, the changes in mRNA levels do not always reflect changes in promoter activity, suggesting that these steroids may have additional post-transcriptional effects on steady-state apoAI mRNA levels. It remains to be established if the transcriptional effects we observed are mediated through an androgen response element. PMID- 11887178 TI - Phosphatidylinositol 3-kinase may mediate isoproterenol-induced vascular relaxation in part through nitric oxide production. AB - Phosphatidylinositol 3-kinase (PI3-K) has been shown to mediate insulin and insulin-like growth factor-1 (IGF-1)-induced nitric oxide (NO) generation and, thus, vascular tone. A role for PI3-K in G-protein-coupled receptor signal transduction has also been reported. As beta2 -adrenergic vascular actions are partly dependent on NO, this study the role of PI3-K on in vitro isoproterenol (Iso)-induced endothelial cell (EC) nitric oxide synthase (NOS) activation and rat aortic vascular relaxation. Cell lysates of rat aortic EC (RAEC), exposed to Iso (10 micromol/L) for 5 minutes, were immunoprecipitated with an antiphosphotyrosine antibody prior to assay for Western blot for the p85-kd regulatory subunit of PI3-K. Endothelial NOS activity was determined by measuring nitrite production. Endothelium-intact aortic rings from male Wistar rats were preincubated with the PI3-K inhibitors, wortmannin (WT), or LY294002 (LY), precontracted with phenylepinephrine (PE), and relaxation to graded doses of Iso was measured. NO contribution to vascular relaxation was assessed by L-N(G) nitroarginine methyl ester (L-NAME), a NOS inhibitor. Both Iso and IGF-1 induced an increase in p85 subunit phosphorylation as demonstrated by Western analysis, effects inhibited by preincubation with WT. Iso also enhanced association of p85 with the Triton X-100-insoluble fraction of RAEC, reflecting translocation of this enzyme to a cytoskeletal fraction. In addition, Iso as well as IGF-1 significantly increased eNOS activity measured by nitrite production. Both WT and LY markedly inhibited relaxation to Iso, while L-NAME nearly abolished this beta adrenergic-mediated vasorelaxation. These data indicate that both Iso and IGF-1 activate the EC PI3-K pathway which mediates, in part, the release of NO and subsequent vasorelaxation in response to this beta-agonist Iso as well as to IGF 1. PMID- 11887179 TI - Efficacy and safety of the new 60-mg formulation of the long-acting somatostatin analog lanreotide in the treatment of acromegaly. AB - Recently, a new slow-release (SR) formulation of lanreotide (LAN) comprising 60 mg of the drug incorporated in microspheres of biodegradable polymers (SR-LAN 60) has become available. The aim of our study was to assess the effectiveness of SR LAN 60, administered every 21 to 28 days, as well as its tolerability in the long term treatment of acromegalic patients treated with SR-LAN 30. Twenty patients with acromegaly (10 males and 10 females) were enrolled in this open study. Thirteen patients had undergone surgery, but with incomplete resection of the pituitary tumor. All patients, treated with intramuscular (IM) SR-LAN 30 injections every 10 days for 12 to 24 months, started SR-LAN 60 (Ipsen-Beaufour, Milan, Italy) administration 10 days after the last injection of SR-LAN 30. Growth hormone (GH) levels were determined on the day of the first injection of SR-LAN 60, and 10, 20, and 30 days after. According to the GH levels reached on day 30, patients received SR-LAN 60 every 28 days if GH levels were below 2.5 microg/L (group A) and every 21 days if GH levels were above 2.5 microg/L (group B). In group A, after the 8th month, SR-LAN 60 treatment resulted in well controlled GH levels in 9 of 10 patients in comparison to SR-LAN 30 treatment every 10 days (6 of 10 patients). Normal age-adjusted insulin-like growth factor I (IGF-I) levels were achieved in 4 of 10 patients, as in treatment with SR-LAN 30. In group B, SR-LAN 60 treatment resulted in well-controlled GH levels in 4 of 10 patients, as in treatment with SR-LAN 30 every 10 days. Normal age-adjusted IGF-I levels were achieved in 3 of 10 patients after SR-LAN 60 in comparison to SR-LAN 30 treatment every 10 days (1 of 10 patients). During SR-LAN 60 therapy, an improvement was also observed in signs and symptoms of active acromegaly and no relevant side effects were detected. In conclusion, this study shows that SR LAN 60 treatment is able to induce a good control of circulating GH and IGF-I levels in most acromegalic patients. The first injections of SR-LAN 60 are very helpful in predicting the optimal long-term injection frequency. Patients on SR LAN 30 can be safely and effectively shifted to SR-LAN 60. PMID- 11887180 TI - A novel mutation in the intron 1 splice donor site of the cholesterol ester transfer protein (CETP) gene as a cause of hyperalphalipoproteinemia. AB - The exchange of cholesterol ester (CE) between lipoproteins occurs through the action of cholesterol ester transfer protein (CETP). The human CETP gene is composed of 16 exons encompassing 25 kbp on chromosome 16q13. The objective of this study was to determine whether a mutation in the CETP gene accounted for severe hyperalphalipoproteinemia in an 80-year-old subject. As a secondary objective, we also investigated the allelic frequency of D442G and Int14A mutation in 224 random Han Chinese subjects. DNA sequence analysis of the CETP gene in the patient revealed a peculiar nucleotide pattern in intron 1. To determine whether this peculiarity results in abnormally spliced mRNA, we used reverse-transcriptase polymerase chain reaction (RT-PCR) to amplify and sequence the patient's cDNA using CETP-specific primers that spanned this splice junction. Both the wild-type and mutant cDNA were detected, and the mutant cDNA showed that its 5'-splice site shifted 4 nucleotides upstream. This change results in a frame shift and premature termination at amino acid residue 22, and thus predicts a markedly truncated protein product. Although this patient did not have either the D442G or Int14A allele, we found that the allelic frequency of D442G in 224 subjects was 4.46%. No subjects had the Int14A allele. In conclusion, a novel intron 1 splice site mutation in the CETP gene in 1 patient with hyperalphalipoproteinemia and D442G allelic frequency of 4.46% was found among a normal population in Taiwan. PMID- 11887182 TI - Local cooperativity in the unfolding of an amyloidogenic variant of human lysozyme. AB - Hydrogen exchange experiments monitored by NMR and mass spectrometry reveal that the amyloidogenic D67H mutation in human lysozyme significantly reduces the stability of the beta-domain and the adjacent C-helix in the native structure. In addition, mass spectrometric data reveal that transient unfolding of these regions occurs with a high degree of cooperativity. This behavior results in the occasional population of a partially structured intermediate in which the three alpha-helices that form the core of the alpha-domain still have native-like structure, whereas the beta-domain and C-helix are simultaneously substantially unfolded. This finding suggests that the extensive intermolecular interactions that will be possible in such a species are likely to initiate the aggregation events that ultimately lead to the formation of the well-defined fibrillar structures observed in the tissues of patients carrying this mutation in the lysozyme gene. PMID- 11887181 TI - Decreased allergic lung inflammatory cell egression and increased susceptibility to asphyxiation in MMP2-deficiency. AB - Clearance of recruited immune cells is necessary to resolve inflammatory reactions. We show here that matrix metalloproteinase 2 (MMP2), as part of an interleukin 13 (IL-13)-dependent regulatory loop, dampens inflammation by promoting the egress of inflammatory cells into the airway lumen. MMP2-/- mice showed a robust asthma phenotype and increased susceptibility to asphyxiation induced by allergens. However, whereas the lack of MMP2 reduced the influx of cells into bronchoalveolar lavage (BAL), numerous inflammatory cells accumulated in the lung parenchyma. BAL of MMP2-/- mice lacked normal chemotactic activity, whereas lung inflammatory cells from the same mice showed appropriate chemotactic responses. Thus, MMP2 establishes the chemotactic gradient required for egression of lung inflammatory cells and prevention of lethal asphyxiation. PMID- 11887183 TI - Protein unfolding by the mitochondrial membrane potential. AB - Mitochondria can unfold importing precursor proteins by unraveling them from their N-termini. However, how this unraveling is induced is not known. Two candidates for the unfolding activity are the electrical potential across the inner mitochondrial membrane and mitochondrial Hsp70 in the matrix. Here, we propose that many precursors are unfolded by the electrical potential acting directly on positively charged amino acid side chains in the targeting sequences. Only precursor proteins with targeting sequences that are long enough to reach the matrix at the initial interaction with the import machinery are unfolded by mitochondrial Hsp70, and this unfolding occurs even in the absence of a membrane potential. PMID- 11887184 TI - Phosphorylation of linker histones regulates ATP-dependent chromatin remodeling enzymes. AB - Members of the ATP-dependent family of chromatin remodeling enzymes play key roles in the regulation of transcription, development, DNA repair and cell cycle control. We find that the remodeling activities of the ySWI/SNF, hSWI/SNF, xMi-2 and xACF complexes are nearly abolished by incorporation of linker histones into nucleosomal array substrates. Much of this inhibition is independent of linker histone-induced folding of the arrays. We also find that phosphorylation of the linker histone by Cdc2/Cyclin B kinase can rescue remodeling by ySWI/SNF. These results suggest that linker histones exert a global, genome-wide control over remodeling activities, implicating a new, obligatory coupling between linker histone kinases and ATP-dependent remodeling enzymes. PMID- 11887185 TI - Functional convergence of two lysyl-tRNA synthetases with unrelated topologies. AB - Lysyl-tRNA can be synthesized by both a class I (LysRS-I) and a class II (LysRS II) lysyl-tRNA synthetase. The crystal structure of LysRS-I from Pyrococcus horikoshii at 2.6 A resolution reveals extensive similarity with glutamyl-tRNA synthetase (GluRS). A comparison of the structures of LysRS-I and LysRS-II in complex with lysine shows that both enzymes use similar strategies for substrate recognition within unrelated active site topologies. A docking model based upon the GluRS-tRNA complex suggests how LysRS-I and LysRS-II can recognize the same molecular determinants in tRNALys, as shown by biochemical results, while approaching the acceptor helix of the tRNA from opposite sides. PMID- 11887186 TI - Identification of an organelle receptor for myosin-Va. AB - Little is known about how molecular motors bind to their vesicular cargo. Here we show that myosin-Va, an actin-based vesicle motor, binds to one of its cargoes, the melanosome, by interacting with a receptor-protein complex containing Rab27a and melanophilin, a postulated Rab27a effector. Rab27a binds to the melanosome first and then recruits melanophilin, which in turn recruits myosin-Va. Melanophilin creates this link by binding to Rab27a in a GTP-dependent fashion through its amino terminus, and to myosin-Va through its carboxy terminus. Moreover, this latter interaction, similar to the ability of myosin-Va to colocalize with melanosomes and influence their distribution in vivo, is absolutely dependent on the presence of exon-F, an alternatively spliced exon in the myosin-Va tail. These results provide the first molecular description of an organelle receptor for an actin-based motor, illustrate how alternate exon usage can be used to specify cargo, and further expand the functional repertoire of Rab GTPases and their effectors. PMID- 11887187 TI - Calcium-dependent membrane association sensitizes soluble guanylyl cyclase to nitric oxide. AB - Nitric oxide (NO) is a ubiquitous, cell-permeable intercellular messenger. The current concept assumes that NO diffuses freely through the plasma membrane into the cytoplasm of a target cell, where it activates its cytosolic receptor enzyme, soluble guanylyl cyclase (sGC). Recent evidence, however, suggests that cellular membranes are not only the predominant site of calcium-dependent NO synthesis, but also the site of its distribution and binding. Here we extend this concept to NO signalling to show that active sGC is partially associated with the plasma membrane in a state of enhanced NO sensitivity. After cellular activation, sGC further translocates to the membrane fraction in human platelets and associates with the NO-synthase-containing caveolar fraction in rat lung endothelial cells, in a manner that is dependent on the concentration of intracellular calcium. Our data suggest that the entire NO signalling pathway is more spatially confined than previously assumed and that sGC dynamically translocates to the plasma membrane, where it is sensitized to NO. PMID- 11887188 TI - Therapeutic angiogenesis: the present and the future. PMID- 11887189 TI - Acute effects of maternal smoking on fetal-placental-maternal system hemodynamics. AB - OBJECTIVE: To study acute hemodynamic alterations in the fetal-placental maternal system immediately after maternal exposure to nicotine. METHODS: This is a noncontrolled experimental study involving 21 pregnant smoking women, randomly selected, with uncomplicated pregnancies and without risk factors for fetal heart disease. Patients underwent ultrasound and fetal echocardiography before and after smoking a cigarette. They were asked to abstain from smoking for 12 hours before the study. The mean nicotine content of the cigarettes used in the study was 0.5mg of nicotine and 6mg of carbon monoxide. RESULTS: The average number of cigarettes smoked per a day prior to the study was 9.67. Gestational age ranged between 18 and 36 weeks. The mean maternal heart rate was elevated (P<0.001) as was the mean fetal heart rate (P=0.044). Maternal systolic blood pressure (P=0.004) and diastolic blood pressure (P=0.033) were also elevated after smoking. A decrease occurred in the systolic/diastolic ratio in the right uterine artery (P=0.014) and in the left uterine artery (P=0.039). The other hemodynamic variables remained unchanged. CONCLUSION: Cigarette smoking can cause changes in physiologic variables of fetal-placental circulation, but it does not change fetal cardiac function, in the dose of nicotine and its components used in this study. The decrease in systolic/diastolic ratio in the uterine arteries is probably related to a dose-dependent nicotine pattern. PMID- 11887190 TI - Evaluation of the medical care of patients with hypertension in an emergency department and in ambulatory hypertension unit. AB - OBJECTIVE: To evaluate the characteristics of the patients receiving medical care in the Ambulatory of Hypertension of the Emergency Department, Division of Cardiology, and in the Emergency Unit of the Clinical Hospital of the Ribeirao Preto Medical School. METHODS: Using a protocol, we compared the care of the same hypertensive patients in on different occasions in the 2 different places. The characteristics of 62 patients, 29 men with a mean age of 57 years, were analyzed between January 1996 and December 1997. RESULTS: The care of these patients resulted in different medical treatment regardless of their clinical features and blood pressure levels. Thus, in the Emergency Unit, 97% presented with symptoms, and 64.5% received medication to rapidly reduce blood pressure. In 50% of the cases, nifedipine SL was the elected medication. Patients who applied to the Ambulatory of Hypertension presenting with similar features, or, in some cases, presenting with similar clinically higher levels of blood pressure, were not prescribed medication for a rapid reduction of blood pressure at any of the appointments. CONCLUSION: The therapeutic approach to patients with high blood pressure levels, symptomatic or asymptomatic, was dependent on the place of treatment. In the Emergency Unit, the conduct was, in the majority of cases, to decrease blood pressure immediately, whereas in the Ambulatory of Hypertension, the same levels of blood pressure, in the same individuals, resulted in therapeutic adjustment with nonpharmacological management. These results show the need to reconsider the concept of hypertensive crises and their therapeutical implications. PMID- 11887191 TI - Fontan operation and the cavopulmonary technique: immediate and late results according to the presence of atrial fenestration. AB - OBJECTIVE: To compare immediate and late results in patients with or without fenestration who underwent cavopulmonary anastomosis so that we could assess the efficiency of the technique. METHODS: Sixty-two patients underwent surgery between 1988 and 1999, 41 with fenestration (group I -G I) and 21 without fenestration (group II -G II). Tricuspid atresia was prevalent in group I (23 56%) and single ventricle was prevalent in group II (14-66%). Mean ages at the time of operation were 7.3 years in group I and 7.6 in group II. At late follow up, mean ages were 10.6 years in group I and 12.8 years in group II. RESULTS: Immediate and late mortality were 7.3% in G-I and 4.7% in G-II. Significant pleural effusion occurred in 41.4% of G-I patients and in 23.8% of G-II patients. Significant pericardial effusion occurred in 29.2% and 14.2%, respectively, in groups I and II. Central venous pressure was greater in G-II, 17.7 cm in H2O, as opposed to 15 cm in G-I. Hospital stay was similar between the groups, 26.3 and 21.8 days, respectively. Cyanosis and arterial insaturation occurred in 5 patients, and 4 patients were in functional class II, all from G-I. At late follow-up, 58 (93.5%) were in functional class I. Sinus rhythm was present in 94%, and pulmonary perfusion was similar in both groups. Eleven patients who underwent spirometry had good tolerance to physical effort. CONCLUSION: Atrial fenestration did not improve the immediate or late follow-up of patients who underwent cavopulmonary anastomosis, and is, therefore, dispensable. PMID- 11887192 TI - Does a role exist for tilting-guided therapy in the management of neurocardiogenic syncope? AB - PURPOSE: Upright tilt-table testing (UTT) is an useful method for identifying patients with neurocardiogenic syncope, but its role in the evaluation of therapeutic efficacy is controversial. The aim of this study was to determine the correlation between negative UTT after therapy introduction (acute efficacy) and symptom recurrence during follow-up (chronic efficacy). METHODS: We studied 56 severely symptomatic patients (age 27 +/- 19 years) with recurrent (7 +/-12 episodes) neurocardiogenic syncope (positive UTT). Once empirical pharmacological therapy was initiated, all patients underwent another UTT (therapeutic evaluation test - TET). Therapy was not modified after TET results. The probability of symptom recurrence was analyzed with the Kaplan-Meier method and compared by log rank test in patients with negative and positive TET. RESULTS: Negative UTT after therapy was related to a significantly lower probability of recurrence during follow-up (4.9 versus 52.4% in 12 months, P<0.0001). CONCLUSION: A good correlation exists between acute and long-term efficacy of pharmacological therapy for neurocardiogenic syncope, so that serial UTT may be considered a good method for identifying an effective therapeutic strategy. PMID- 11887194 TI - Autonomic modulation of the heart in systemic arterial hypertension. AB - OBJECTIVE: To analyze the heart rate variability in patients with mild to moderate systemic arterial hypertension. METHODS: Thirty-two healthy (group I) and 70 systemic arterial hypertensive (group II) individuals, divided according to age (40 to 59 and 60 to 80 years old, respectively) and with a similar distribution by sex were studied. Thirty-one had left ventricular hypertrophy (LVH), 22 were overweight, and 16 had Type II diabetes mellitus. Smoking, alcohol ingestion, and sedentary habits were the same between groups. Variability in heart rate was analyzed in the time domain, using standard deviations of normal RR intervals (SDNN) and the differences between maximal brady- and tachycardia (D BTmax) during sustained inspiration. Analysis of the frequency band of the power spectrum between 0.05 and 0.40 Hz at rest and during controlled respiration was chosen for analysis of the frequency domain. RESULTS: In both time and frequency domains, variables were lower in group II than in group I. Within groups, statistically significant variables were only found for individuals in the 40 to 59 year old group. The presence of LVH, overweight, or diabetes mellitus did not influence the variability in heart rate to a significant extent. CONCLUSION: Variability in heart rate was a valuable instrument for analyzing autonomic modulation of the heart in arterial hypertension. The autonomic system undergoes significant losses in cardio-modulatory capacity, more evident in subjects between 40 and 59 years old. In those over 60 years old, reduced variability in heart rate imposed by aging was not significantly influenced by the presence of systemic arterial hypertension. PMID- 11887193 TI - Effects of sibutramine on the treatment of obesity in patients with arterial hypertension. AB - OBJECTIVE: To assess the effects of weight reduction with 10mg of sibutramine or placebo on blood pressure during 24 hours (ambulatory blood pressure monitoring), on left ventricular mass, and on antihypertensive therapy in 86 obese and hypertensive patients for 6 months. METHODS: The patients underwent echocardiography, ambulatory blood pressure monitoring, and measurement of the levels of hepatic enzymes prior to and after treatment with sibutramine or placebo. RESULTS: The group using sibutramine had a greater weight loss than that using placebo (6.7% versus 2.5%; p<0.001), an increase in heart rate (78.3 +/- 7.3 to 82 +/- 7.9 bpm; p=0.02), and a reduction in the left ventricular mass/height index (105 +/- 29.3 versus 96.6 +/- 28.58 g/m; p=0.002). Both groups showed similar increases in the levels of alkaline phosphatase and comparable adjustments in antihypertensive therapy; blood pressure, however, did not change. CONCLUSION: The use of sibutramine caused weight loss and a reduction in left ventricular mass in obese and hypertensive patients with no interference with blood pressure or with antihypertensive therapy. PMID- 11887195 TI - Clinical meaning of ascites in patients with endomyocardial fibrosis. AB - OBJECTIVE: To evaluate the clinical meaning of ascites and the main features of patients with ascites and endomyocardial fibrosis. METHODS: We studied 166 patients with endomyocardial fibrosis (mean age 37 years, 114 women) treated over the last 20 years. Ventriculography findings, surgery or necropsy confirmed the diagnosis in all patients. Most patients belonged to New York Heart Association Functional Class III/IV (134, 83.7%). Eighty-one (50.6%) had biventricular, 28 (17.5%) had right ventricular, and 51 (31.8%) had left ventricular involvement. During follow-up, 56 patients died. RESULTS: Ascites was present in 67 (41.8%) patients, and right ventricular involvement was present in 59 (88%). In the comparison between patients with or without ascites, those with ascites had higher mortality (49.2% and 24.7%, respectively). Patients with ascites had a higher incidence of edema (95% vs. 43%), hepatomegaly (5.8cm vs. 4.1cm), mean right atrium pressure (19.3 vs. 12mmHg), and final right ventricle diastolic pressure (18.7 vs. 12.9mmHg). Also, patients with ascites had a longer history of illness (5.1 and 3.9 years, respectively) and had atrial fibrillation more frequently (44.7% vs. 30.1%). CONCLUSION: Ascites was observed in less than 50% of cases of endomyocardial fibrosis and was associated with greater involvement of the right ventricle and with a longer duration of the disease, thus being a characteristic of a worse prognosis. PMID- 11887196 TI - Study of the myocardial contraction and relaxation velocities through Doppler tissue imaging echocardiography: A new alternative in the assessment of the segmental ventricular function. AB - OBJECTIVE: Doppler tissue imaging (DTI) enables the study of the velocity of contraction and relaxation of myocardial segments. We established standards for the peak velocity of the different myocardial segments of the left ventricle in systole and diastole, and correlated them with the electrocardiogram. METHODS: We studied 35 healthy individuals (27 were male) with ages ranging from 12 to 59 years (32.9 plus minus 10.6). Systolic and diastolic peak velocities were assessed by Doppler tissue imaging in 12 segments of the left ventricle, establishing their mean values and the temporal correlation with the cardiac cycle. RESULTS: The means (and standard deviation) of the peak velocities in the basal, medial, and apical regions (of the septal, anterior, lateral, and posterior left ventricle walls) were respectively, in cm/s, 7.35(1.64), 5.26(1.88), and 3.33(1.58) in systole and 10.56(2.34), 7.92(2.37), and 3.98(1.64) in diastole. The mean time in which systolic peak velocity was recorded was 131.59ms (+/- 19.12ms), and diastolic was 459.18ms (+/- 18.13ms) based on the peak of the R wave of the electrocardiogram. CONCLUSION: In healthy individuals, maximum left ventricle segment velocities decreased from the bases to the ventricular apex, with certain proportionality between contraction and relaxation (P<0.05). The use of Doppler tissue imaging may be very helpful in detecting early alterations in ventricular contraction and relaxation. PMID- 11887197 TI - Influence of quality of sleep on the nocturnal decline in blood pressure during ambulatory blood pressure monitoring. AB - OBJECTIVE: To assess the influence of the quality of sleep on the nocturnal physiological drop in blood pressure during ambulatory blood pressure monitoring. METHODS: We consecutively assessed ambulatory blood pressure monitoring, the degree of tolerance for the examination, and the quality of sleep in 168 patients with hypertension or with the suspected "white-coat" effect. Blood pressure fall during sleep associated with a specific questionnaire and an analogical visual scale of tolerance for ambulatory blood pressure monitoring were used to assess usual sleep and sleep on the day of examination. Two specialists in sleep disturbances classified the patients into 2 groups: those with normal sleep and those with abnormal sleep. RESULTS: Fifty-nine (35 %) patients comprised the abnormal sleep group. Findings regarding the quality of sleep on the day of ambulatory blood pressure monitoring as compared with those regarding the quality of sleep on a usual day were different and were as follows, respectively: total duration of sleep (-12.4 +/- 4.7 versus -42.2 +/- 14.9 minutes, P=0.02), latency of sleep (0.4 +/- 2.7 versus 17 +/- 5.1 minutes, P<0.001), number of awakenings (0.1 +/- 0.1 versus 1.35 +/- 0.3 times, P<0.001), and tolerance for ambulatory blood pressure monitoring (8 +/- 0.2 versus 6.7 +/- 0.35, P=0.035). An abnormal drop in blood pressure during sleep occurred in 20 (18%) patients in the normal sleep group and in 14 (24%) patients in the abnormal sleep group, P=0.53. CONCLUSION: Ambulatory blood pressure monitoring causes sleep disturbances in some patients, and a positive association between quality of sleep and tolerance for the examination was observed. PMID- 11887198 TI - Mitral valve replacement and remodeling of the left ventricle in dilated cardiomyopathy with mitral regurgitation: initial results. AB - OBJECTIVE: This study evaluated the effects of a new method of mitral valve replacement on left ventricular (LV) remodeling and heart failure functional class. METHODS: Eight patients (6 men) with severe mitral regurgitation from end stage dilated cardiomyopathy underwent surgery. Five patients were in functional class (FC) IV, 2 were in FC III and 1 was in FC III/IV. Age ranged from 33 to 63 years. Both the anterior and posterior leaflets of the mitral valve were divided into hemileaflets. The resultant 4 pedicles were displaced under traction toward the left atrium and anchored between the mitral annulus and an implanted valvular prosthesis. The beating heart facilitated ideal chordae tendineae positioning. RESULTS: All patients survived and were discharged from the hospital. After a mean follow-up period of 6.5 months (1-12 m), 5 patients were in FC I; 2 in FC I/II; and 1 in FC II. The preoperative ejection fraction ranged from 19% to 30% (mean: 25.7 +/- 3.4 %), and the postoperative ejection fraction ranged from 21% to 40% (mean: 31.1 +/- 5.8%). Doppler echocardiography showed evidence of LV remodeling in 4 patients, including lateral wall changes and a tendency of the LV cavity to return to its elliptical shape. CONCLUSION: This technique of mitral valve replacement, involving new positioning of the chordae tendineae, allowed LV remodeling and improvement in FC during this brief follow-up period. PMID- 11887199 TI - Low back pain during streptokinase infusion. AB - We report the case of a 57-year-old male patient with severe low back pain during streptokinase infusion administered to treat typical chest pain and elevation of the ST segment in the inferior wall. We reviewed the literature, emphasizing the differential diagnosis, the pathophysiology, and management of the event. PMID- 11887200 TI - Aneurysm of the right atrial appendage. AB - Atrial aneurysms involving the free wall or atrial appendage are rare entities in cardiology practice and may be associated with atrial arrhythmias or embolic phenomena. We review the literature and report a case of aneurysm of the right atrial appendage in a young adult, whose diagnosis was established with echocardiography after an episode of paroxysmal atrial flutter. PMID- 11887201 TI - [Anatomoclinical correlation. Case 1/2002 - Uncompensated heart failure and ascites in a 51-year-old female with a biological mitral prosthesis]. PMID- 11887202 TI - [Clinic-radiographic correlation. Case 2/2002 - Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 11887203 TI - [Evolution of the percutaneous intervention for the treatment of multiarterial coronary disease]. PMID- 11887204 TI - Estrogen replacement therapy and cardioprotection: mechanisms and controversies. AB - Epidemiological and case-controlled studies suggest that estrogen replacement therapy might be beneficial in terms of primary prevention of coronary heart disease (CHD). This beneficial effect of estrogens was initially considered to be due to the reduction of low density lipoproteins (LDL) and to increases in high density lipoproteins (HDL). Recent studies have shown that estrogens protect against oxidative stress and decrease LDL oxidation. Estrogens have direct effects on the arterial tissue and modulate vascular reactivity through nitric oxide and prostaglandin synthesis. While many of the effects of estrogen on vascular tissue are believed to be mediated by estrogen receptors alpha and beta, there is evidence for 'immediate non-genomic' effects. The role of HDL in interacting with 17beta-estradiol including its esterification and transfer of esterified estrogens to LDL is beginning to be elucidated. Despite the suggested positive effects of estrogens, two recent placebo-controlled clinical trials in women with CHD did not detect any beneficial effects on overall coronary events with estrogen therapy. In fact, there was an increase in CHD events in some women. Mutations in thrombogenic genes (factor V Leiden, prothrombin mutation, etc.) in a subset of women may play a role in this unexpected finding. Thus, the cardioprotective effect of estrogens appears to be more complicated than originally thought and requires more research. PMID- 11887205 TI - Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8. AB - SDS, C12E8, CHAPS or CHAPSO or a combination of two of these detergents is generally used for the solubilization of Na,K-ATPase and other ATPases. Our method using only C12E8 has the advantage of considerable reduction of the time for enzyme purification, with rapid solubilization and purification in a single chromatographic step. Na,K-ATPase-rich membrane fragments of rabbit kidney outer medulla were obtained without adding SDS. Optimum conditions for solubilization were obtained at 4 degrees C after rapid mixing of 1 mg of membrane Na,K-ATPase with 1 mg of C12E8/ml, yielding 98% recovery of the activity. The solubilized enzyme was purified by gel filtration on a Sepharose 6B column at 4 degrees C. Non-denaturing PAGE revealed a single protein band with phosphomonohydrolase activity. The molecular mass of the purified enzyme estimated by gel filtration chromatography was 320 kDa. The optimum apparent pH obtained for the purified enzyme was 7.5 for both PNPP and ATP. The dependence of ATPase activity on ATP concentration showed high (K0.5 = 4.0 microM) and low (K0.5 = 1.4 mM) affinity sites for ATP, with negative cooperativity. Ouabain (5 mM), oligomycin (1 microg/ml) and sodium vanadate (3 microM) inhibited the ATPase activity of C12E8 solubilized and purified Na,K-ATPase by 99, 81 and 98.5%, respectively. We have shown that Na,K-ATPase solubilized only with C12E8 can be purified and retains its activity. The activity is consistent with the form of (alphabeta)2 association. PMID- 11887206 TI - Effect of iodide on Fas, Fas-ligand and Bcl-w mRNA expression in thyroid of NOD mice pretreated with methimazole. AB - Nonobese diabetic (NOD) mice and a derived strain, NOD.H.2h4, have been used as a model for experimental spontaneous thyroiditis and thyroiditis induced by iodide excess after a goiter-inducing period. Some authors have proposed that iodide, given after methimazole or propylthiouracil, is capable of inducing apoptosis in thyroid cells and that anti-thyroid drugs can modulate the expression of apoptosis components such as Fas and its ligand (Fas-L). Here we evaluated the effect of potassium iodide (20 microg/animal for 4 days, i.p.) given to NOD mice at the 10th week of life after exposure to methimazole (1 mg/ml) in drinking water from the 4th to the 10th week of life. Fas, Fas-L and Bcl-w expression were analyzed semiquantitatively by RT-PCR immediately after potassium iodide administration (group MI44D) or at week 32 (MI32S). Control groups were added at 10 (C10) and 32 weeks (C32), as well as a group that received only methimazole (CM10). An increase in the expression of Fas-L and Bcl-w (P<0.01, ANOVA) was observed in animals of group MI44D, while Fas was expressed at higher levels (P = 0.02) in group C32 (72.89 +/- 47.09 arbitrary units) when compared to group C10 (10.8 +/- 8.55 arbitrary units). Thus, the analysis of Fas-L and Bcl-w expression in the MI44D group and Fas in group C32 allowed us to detect two different patterns of expression of these apoptosis components in thyroid tissue of NOD mice. PMID- 11887208 TI - Performance of a Brazilian population sample in the Boston Diagnostic Aphasia Examination: a pilot study. AB - Brazilian researchers and health professionals often face the challenge of having to use tests developed in foreign languages and standardized for populations of other countries, especially in the fields of Neuropsychology and Neurolinguistics. This fact promotes a feeling that some scoring systems may be inadequate for our sociocultural reality. In the present study, we describe the performance of a Brazilian population sample submitted to a translated and adapted version of the Boston Diagnostic Aphasia Examination (BDAE). Sixty normal volunteers (21 men and 39 women), all Portuguese native speakers, ranging in age from 15 to 78 years (average 43.7) and with an educational level of 2 to 16 years (average 9.9), were tested using a translated and adapted Portuguese version of the BDAE. Cut-off scores are suggested for our population and the performance of the Brazilian sample is compared to that of American and Colombian samples, with the results being closely similar in all tasks. We also performed a correlation analysis between age, gender and educational level and the influence of these variables on the performance of the subjects. We found no statistically significant differences between genders. Educational level correlated positively with performance, especially in the subtests involving reading and writing. There was a negative correlation between age and performance in two subtests (Visual Confrontation Naming and Sentences to Dictation), but a coexisting effect of educational level could not be ruled out. PMID- 11887207 TI - The leaves of green plants as well as a cyanobacterium, a red alga, and fungi contain insulin-like antigens. AB - We report the detection of insulin-like antigens in a large range of species utilizing a modified ELISA plate assay and Western blotting. We tested the leaves or aerial parts of species of Rhodophyta (red alga), Bryophyta (mosses), Psilophyta (whisk ferns), Lycopodophyta (club mosses), Sphenopsida (horsetails), gymnosperms, and angiosperms, including monocots and dicots. We also studied species of fungi and a cyanobacterium, Spirulina maxima. The wide distribution of insulin-like antigens, which in some cases present the same electrophoretic mobility as bovine insulin, together with results recently published by us on the amino acid sequence of an insulin isolated from the seed coat of jack bean (Canavalia ensiformis) and from the developing fruits of cowpea (Vigna unguiculata), suggests that pathways depending on this hormone have been conserved through evolution. PMID- 11887209 TI - Effect of asthma severity on symptom perception in childhood asthma. AB - Individual ability to perceive airway obstruction varies substantially. The factors influencing the perception of asthma are probably numerous and not well established in children. The present study was designed to examine the influence of asthma severity, use of preventive medication, age and gender on the association between respiratory symptoms (RS) and peak expiratory flow (PEF) rates in asthmatic children. We followed 92 asthmatic children, aged 6 to 16 years, for five months. Symptom scores were recorded daily and PEF was measured twice a day. The correlations among variables at the within-person level over time were analyzed for each child and for the pooled data by multivariate analysis. After pooling the data, there was a significant (P<0.05) correlation between each symptom and PEF; 60% of the children were accurate perceivers (defined by a statistically significant correlation between symptoms and PEF across time) for diurnal symptoms and 37% for nocturnal symptoms. The accuracy of perception was independent of asthma severity, age, gender or the use of preventive medication. Symptom perception is inaccurate in a substantial number of asthmatic children, independently of clinical severity, age, gender or use of preventive medication. It is not clear why some asthmatic patients are capable of accurately perceiving the severity of airway obstruction while others are not. PMID- 11887210 TI - Analysis of HLA-A antigens and C282Y and H63D mutations of the HFE gene in Brazilian patients with hemochromatosis. AB - The hemochromatosis gene, HFE, is located on chromosome 6 in close proximity to the HLA-A locus. Most Caucasian patients with hereditary hemochromatosis (HH) are homozygous for HLA-A3 and for the C282Y mutation of the HFE gene, while a minority are compound heterozygotes for C282Y and H63D. The prevalence of these mutations in non-Caucasian patients with HH is lower than expected. The objective of the present study was to evaluate the frequencies of HLA-A antigens and the C282Y and H63D mutations of the HFE gene in Brazilian patients with HH and to compare clinical and laboratory profiles of C282Y-positive and -negative patients with HH. The frequencies of HLA-A and C282Y and H63D mutations were determined by PCR-based methods in 15 male patients (median age 44 (20-72) years) with HH. Eight patients (53%) were homozygous and one (7%) was heterozygous for the C282Y mutation. None had compound heterozygosity for C282Y and H63D mutations. All but three C282Y homozygotes were positive for HLA-A3 and three other patients without C282Y were shown to be either heterozygous (N = 2) or homozygous (N = 1) for HLA A3. Patients homozygous for the C282Y mutation had higher ferritin levels and lower age at onset, but the difference was not significant. The presence of C282Y homozygosity in roughly half of the Brazilian patients with HH, together with the findings of HLA-A homozygosity in C282Y-negative subjects, suggest that other mutations in the HFE gene or in other genes involved in iron homeostasis might also be linked to HH in Brazil. PMID- 11887211 TI - Comparison of methods for urinary albumin determination in patients with type 1 diabetes. AB - We tested the correlation of the albumin-to-creatinine ratio (A/C) in an early morning urine sample, measured with a commercial kit (DCA 2000), with the conventional immunoturbidimetric determination in the laboratory and with overnight albumin excretion rate (reference method). Fifty-five type 1 diabetic adolescents had their first-morning urine collected on the 1st and 8th day of the period. Urinary albumin and creatinine were determined immediately using the DCA 2000 kit. Samples were also stored for laboratory analysis. To evaluate the correlation between early-morning urinary A/C ratio and overnight albumin excretion rate, 16 subjects had a timed overnight urine collection. A/C ratios determined with the DCA 2000 kit and by the laboratory method were 13.1 +/- 20.5 and 20.4 +/- 46.3 mg/g, respectively. A/C results by both methods proved to be strongly correlated (r = 0.98, P<0.001). DCA 2000-determined A/C showed 50% sensitivity and 100% specificity when compared to the reference method. Spot urinary A/C of the subset of 16 subjects significantly correlated with their overnight albumin excretion rate (r = 0.98, P<0.001). Intraindividual variation ranged from 17 to 32% and from 9 to 63% for A/C and overnight albumin excretion rate, respectively. In conclusion, an early-morning specimen should be used instead of timed overnight urine and the A/C ratio is an accurate, reliable and easily determined parameter for the screening of diabetic nephropathy. Immediate measurement of the A/C ratio is feasible using the DCA 2000 kit. Intraindividual variability indicates the need for repeated determinations to confirm microalbuminuria and the diagnosis of incipient diabetic nephropathy. PMID- 11887212 TI - Determination of serum aluminum, platelet aggregation and lipid peroxidation in hemodialyzed patients. AB - Aluminum (Al3+) overload is frequently associated with lipid peroxidation and neurological disorders. Aluminum accumulation is also reported to be related to renal impairment, anemia and other clinical complications in hemodialysis patients. The aim of the present study was to determine the degree of lipid peroxidation, platelet aggregation and serum aluminum in patients receiving regular hemodialytic treatment. The level of plasma lipid peroxidation was evaluated on the basis of thiobarbituric acid reactive substances (TBARS). Mean platelet peroxidation in patients undergoing hemodialysis was significantly higher than in normal controls (2.7 +/- 0.03 vs. 1.8 +/- 0.06 nmol/l, P<0.05). Platelet aggregation and serum aluminum levels were determined by a turbidimetric method and atomic absorption spectrophotometry, respectively. Serum aluminum was significantly higher in patients than in normal controls (44.5 +/- 29 vs. 10.8 +/ 2.5 microg/l, P<0.05). Human blood platelets were stimulated with collagen (2.2 microg/ml), adenosine diphosphate (6 microM) and epinephrine (6 microM) and showed reduced function with the three agonists utilized. No correlation between aluminum levels and platelet aggregation or between aluminum and peroxidation was observed in hemodialyzed patients. PMID- 11887213 TI - Aberrant crypt foci and colon cancer: comparison between a short- and medium-term bioassay for colon carcinogenesis using dimethylhydrazine in Wistar rats. AB - Aberrant crypt foci (ACF) in the colon of carcinogen-treated rodents are considered to be the earliest hallmark of colon carcinogenesis. In the present study the relationship between a short-term (4 weeks) and medium-term (30 weeks) assay was assessed in a model of colon carcinogenesis induced by dimethylhydrazine (DMH) in the rat. Six-week-old male Wistar rats were given subcutaneous injections of DMH (40 mg/kg) twice a week for 2 weeks and killed at the end of the 4th or 30th week. ACF were scored for number, distribution pattern along the colon and crypt multiplicity in 0.1% methylene-blue whole-mount preparations. ACF were distinguished from normal crypts by their larger size and elliptical shape. The incidence, distribution and morphology of colon tumors were recorded. The majority of ACF were present in the middle and distal colon of DMH treated rats and their number increased with time. By the 4th week, 91.5% ACF were composed of one or two crypts and 8.5% had three or more crypts, while by the 30th week 46.9% ACF had three or more crypts. Thus, a progression of ACF consisting of multiple crypts was observed from the 4th to the 30th week. Nine well-differentiated adenocarcinomas were found in 10 rats by the 30th week. Seven tumors were located in the distal colon and two in the middle colon. No tumor was found in the proximal colon. The present data indicate that induction of ACF by DMH in the short-term (4 weeks) assay was correlated with development of well differentiated adenocarcinomas in the medium-term (30 weeks) assay. PMID- 11887214 TI - Nephrotoxicity of Bence-Jones proteins: interference in renal epithelial cell acidification. AB - The aim of the present study was to evaluate the acidification of the endosome lysosome system of renal epithelial cells after endocytosis of two human immunoglobulin lambda light chains (Bence-Jones proteins, BJP) obtained from patients with multiple myeloma. Renal epithelial cell handling of two BJP (neutral and acidic BJP) was evaluated by rhodamine fluorescence. Renal cells (MDCK) were maintained in culture and, when confluent, were incubated with rhodamine-labeled BJP for different periods of time. Photos were obtained with a fluorescence microscope (Axiolab-Zeiss). Labeling density was determined on slides with a densitometer (Shimadzu Dual-Wavelength Flying-Spot Scanner CS9000). Endocytosis of neutral and acidic BJP was correlated with acidic intracellular compartment distribution using acridine orange labeling. We compared the pattern of distribution after incubation of native neutral and acidic BJP and after complete deglycosylation of BJP by periodate oxidation. The subsequent alteration of pI converted neutral BJP to acidic BJP. There was a significant accumulation of neutral BJP in endocytic structures, reduced lysosomal acidification, and a diffuse pattern of acidification. This pattern was reversed after total deglycosylation and subsequent alteration of the pI to an acidic BJP. We conclude that the physicochemical characteristics of BJP interfere with intracellular acidification, possibly explaining the strong nephrotoxicity of neutral BJP. Lysosomal acidification is fundamental for adequate protein processing and catabolism. PMID- 11887215 TI - Low pH and calcium effects on net Na+ and K+ fluxes in two catfish species from the Amazon River (Corydoras: Callichthyidae). AB - The present study analyzes Na+ and K+ disturbances caused by low pH in two catfish species from the Amazon River. Corydoras adolfoi inhabits ion-poor, black stained, low pH (3.5-4.0) waters, while C. schwartzi is native to ion-rich waters at circumneutral pH. Fish were exposed to pH 3.5 Ca2+-free, and Ca2+-enriched (approximately 500 micromol/l) water to determine the protective effects of calcium. Net Na+ and K+ fluxes were measured in the water collected from the fish experimental chambers. C. adolfoi was unable to control the Na+ efflux at low pH, exhibiting Na+ loss up to -594 +/- 84 nmol g(-1) h(-1) during the first hour. After 3 and 6 h, net Na+ flux increased by 7- and 23-fold, respectively. In C. schwartzi, at pH 3.5, the initial high Na+ loss (-1,063 +/- 73 nmol g(-1) h(-1)) was gradually attenuated. A K+ loss occurred in both species, but remained relatively constant throughout exposure. High [Ca2+] affected ion losses in both species. C. adolfoi had 70% loss attenuation, indicating incapacity to control Na+ efflux. In C. schwartzi, elevated [Ca2+] completely prevented the Na+ losses caused by exposure to low pH. Rather different patterns were seen for K+ fluxes, with C. adolfoi showing no K+ disruption when exposed to low pH/high [Ca2+]. Thus, C. adolfoi loses Na+ during acid exposure, but has the ability to control K+ loss, while C. schwartzi controls diffusive Na+ loss but exhibits a slightly higher K+ loss. Ion balance was influenced by [Ca2+] at low pH in C. schwartzi but not in C. adolfoi. PMID- 11887216 TI - Serotyping HIV-1 with V3 peptides: detection of high avidity antibodies presenting clade-specific reactivity. AB - The main objective of the present study was to assess the specificity and sensitivity of a modified assay using short synthetic peptides of the V3 region of HIV-1 gp120, which is the main target for neutralizing antibodies. Results from an enzyme immunoassay (EIA) employing a panel of synthetic peptides of HIV-1 subtypes and using urea washes to detect high avidity antibodies (AAV3) were compared with those obtained by the heteroduplex mobility assay and DNA sequencing. The EIA correctly typed 100% of subtype B (sensitivity = 1.0; specificity = 0.95), 100% of HIV-1 E samples (sensitivity = 1.0; specificity = 1.0), and 95% of subtype C specimens (sensitivity = 0.95; specificity = 0.94). In contrast, only 50% of subtype A (sensitivity = 0.5; specificity = 0.95), 60% of subtype D (sensitivity = 0.6; specificity = 1.0), and 28% of subtype F samples (sensitivity = 0.28; specificity = 0.95) were correctly identified. This approach was also able to discriminate in a few samples antibodies from patients infected with B variants circulating in Brazil and Thailand that reacted specifically. The assays described in this study are relatively rapid and simple to perform compared to molecular approaches and can be used to screen large numbers of serum or plasma samples. Moreover, the classification in subtypes (genotypes) may overestimate HIV-1 diversity and a classification into serotypes, based on antigenic V3 diversity or another principal neutralization domain, may be more helpful for vaccine development and identification of variants. PMID- 11887217 TI - Sm14 gene expression in different stages of the Schistosoma mansoni life cycle and immunolocalization of the Sm14 protein within the adult worm. AB - Sm14 is a 14-kDa vaccine candidate antigen from Schistosoma mansoni that seems to be involved in cytoplasmic trafficking of fatty acids. Although schistosomes have a high requirement for lipids, they are not able to synthesize fatty acids and sterols de novo. Thus, they must acquire host lipids. In order to determine whether Sm14 is present in different stages of the life cycle of the parasite, we performed RT-PCR. Sm14 mRNA was identified in all stages of the life cycle studied, mainly schistosomulum, adult worm and egg. Additionally, we used a rabbit anti-Sm14 polyclonal antibody in an indirect immunofluorescence assay to localize Sm14 in adult worm sections. The basal lamella of the tegument and the gut epithelium were strongly labeled. These tissues have a high flow of and demand for lipids, a finding that supports the putative role of Sm14 as an intracellular transporter of fatty acids from host cells. PMID- 11887218 TI - Phagocytosis by macrophages mediated by receptors for denatured proteins - dependence on tyrosine protein kinases. AB - Previous studies have demonstrated that some components of the leukocyte cell membrane, CR3 (Mac-1, CD11b/CD18) and p150/95, are able to bind to denatured proteins. Thus, it is of interest to know which effector functions of these cells can be triggered by these receptors when they interact with particles or surfaces covered with denatured proteins. In the present study we analyzed their possible role as mediators of phagocytosis of red cells covered with denatured bovine serum albumin (BSA) by mouse peritoneal macrophages. We observed that a) macrophages are able to recognize (bind to) these red cells, b) this interaction can be inhibited by denatured BSA in the fluid phase, c) there is no phagocytosis of these particles by normal macrophages, d) phagocytosis mediated by denatured BSA can be, however, effectively triggered in inflammatory macrophages induced by glycogen or in macrophages activated in vivo with LPS, and e) this phagocytic capacity is strongly dependent on the activity of tyrosine protein kinases in its signal transduction pathway, as demonstrated by using three kinds of enzyme inhibitors (genistein, quercetin and herbimycin A). PMID- 11887219 TI - Reproductive experience influences grooming behavior during pregnancy in rats. AB - The pregnancy-induced increase in self-licking observed in rats is important for mammary gland development and lactation. Reproductive experience has epidemiologial implications such as a decrease in the incidence of mammary gland cancer in women and it also influences various behavioral, neurochemical and endocrine parameters. The aim of the present study was to investigate the influence of reproductive experience on grooming behavior patterns during pregnancy in rats. Self-grooming behavior was measured in age-matched virgin, primi- and multigravid (days 7, 8, 9, 19, and 20 of pregnancy) rats. General grooming (head, forelimbs and shoulders) was not significantly different among virgin, primi- and multigravid rats during pregnancy. Confirming previous work, pregnant rats spent significantly more time in specific grooming (mammary glands, nipple lines, genital and pelvic regions) than did virgin animals. In addition, self- licking of mammary glands was significantly increased in multi- as compared to primigravid rats on days 8, 9, 19 and 20 of pregnancy. The increase in mammary gland grooming observed in multigravid rats appears to be a consequence of previous reproductive experience. These data show that reproductive experience modulates mammary gland grooming during pregnancy, possibly contributing to successful reproduction. PMID- 11887220 TI - Antinociceptive potency of aminoglycoside antibiotics and magnesium chloride: a comparative study on models of phasic and incisional pain in rats. AB - A close relationship exists between calcium concentration in the central nervous system and nociceptive processing. Aminoglycoside antibiotics and magnesium interact with N- and P/Q-type voltage-operated calcium channels. In the present study we compare the antinociceptive potency of intrathecal administration of aminoglycoside antibiotics and magnesium chloride in the tail-flick test and on incisional pain in rats, taken as models of phasic and persistent post-surgical pain, respectively. The order of potency in the tail-flick test was gentamicin (ED50 = 3.34 microg; confidence limits 2.65 and 4.2) > streptomycin (5.68 microg; 3.76 and 8.57) = neomycin (9.22 microg; 6.98 and 12.17) > magnesium (19.49 microg; 11.46 and 33.13). The order of potency to reduce incisional pain was gentamicin (ED50 = 2.06 microg; confidence limits 1.46 and 2.9) > streptomycin (47.86 microg; 26.3 and 87.1) = neomycin (83.17 microg; 51.6 and 133.9). The dose response curves for each test did not deviate significantly from parallelism. We conclude that neomycin and streptomycin are more potent against phasic pain than against persistent pain, whereas gentamicin is equipotent against both types of pain. Magnesium was less potent than the antibiotics and effective in the tail flick test only. PMID- 11887221 TI - Variations in gastric compliance induced by acute blood volume changes in anesthetized rats. AB - The impact of acute volume imbalances on gastric volume (GV) was studied in anesthetized rats (250-300 g). After cervical and femoral vessel cannulation, a balloon catheter was positioned in the proximal stomach. The opposite end of the catheter was connected to a barostat with an electronic sensor coupled to a plethysmometer. A standard ionic solution was used to fill the balloon (about 3.0 ml) and the communicating vessel system, and to raise the reservoir liquid level 4 cm above the animals' xiphoid appendix. Due to constant barostat pressure, GV values were considered to represent the gastric compliance index. All animals were monitored for 90 min. After a basal interval, they were randomly assigned to normovolemic, hypervolemic, hypovolemic or restored protocols. Data were compared by ANOVA followed by Bonferroni's test. Mean arterial pressure (MAP), central venous pressure (CVP) and GV values did not change in normovolemic animals (N = 5). Hypervolemic animals (N = 12) were transfused at 0.5 ml/min with a suspension of red blood cells in Ringer-lactate solution with albumin (12.5 ml/kg), which reduced GV values by 11.3% (P<0.05). Hypovolemic rats (N = 12) were bled up to 10 ml/kg, a procedure that increased GV values by 15.8% (P<0.05). In the restored group (N = 12), shed blood replacement brought GV values back to basal levels in bled animals (P>0.05). MAP and CVP values increased (P<0.05) after hypervolemia but decreased (P<0.05) with hypovolemia. In conclusion, blood volume level modulates gastric compliance, turning the stomach into an adjustable reservoir, which could be part of the homeostatic process to balance blood volume. PMID- 11887222 TI - [Non-transmissible diseases and infections]. PMID- 11887223 TI - [Weight excess and abdominal fat in the metabolic syndrome among Japanese Brazilians]. AB - OBJECTIVE: Obesity, especially abdominal, has been associated with cardiovascular risk factors such as dyslipidemia, hypertension and diabetes mellitus (DM). The importance of these risk factors among Japanese-Brazilians was previously shown, although obesity is not a typical characteristic of Japanese migrants. In this study the prevalence of weight excess and central adiposity (CA) among Japanese Brazilians and their association with metabolic disorders was evaluated. METHODS: A sample of 530 1st and 2nd generation Japanese-Brazilians (aged 40 - 79 years) went through anthropometric and blood pressure measurements, lipid profile and oral glucose tolerance tests. The prevalence rate (point and confidence interval) of overweight was calculated using a cut-off value of >26.4 kg/m2. CA diagnosis was based on waist-to-hip circumference ratio (WHR): greater-than-or-equal 0.85 and 0.95 in women and men, respectively. RESULTS: The prevalence of weight excess was 22.4% (CI 95% 20.6 - 28.1), and CA was 67.0% (95% CI 63.1 - 70.9). In addition to higher prevalence of DM, hypertension and dyslipidemia, stratifying by BMI and WHR, people with weight excess and CA revealed a poorer metabolic profile: blood pressure levels were significantly higher among those with weight excess with or without CA; CA individuals had higher glucose, triglycerides, total and LDL cholesterol, and lower HDL than those without weight excess or CA; fasting insulinemia was significantly higher among subjects with weight excess (with or without CA) than among those without weight excess or CA. CONCLUSION: Comparing subgroups with and without CA supports the hypothesis that abdominal fat accumulation represents a risk factor for insulin resistance-related diseases, even among Japanese descendants. The increased prevalence of metabolic syndrome among Japanese migrants could be attributed to visceral fat deposition, which has been implicated in the genesis of insulin resistance. PMID- 11887224 TI - [Food consumption scores and serum lipids levels in the population of Sao Paulo, Brazil]. AB - OBJECTIVE: To study food patterns assessed using scores of consumption and their relationship with serum total cholesterol (TOTAL-C), low density lipoprotein cholesterol (LDL-C) and high density lipoproteins (HDL-C) concentration in the population of the metropolitan area of Sao Paulo, Brazil. METHODS: Data on food frequency consumption (FFC), serum lipids, and other covariates were available for a representative sample of 1,045 adults. A 12-month retrospective food frequency questionnaire was used. FFC was assessed using scores of consumption, which was obtained by grouping food according to their composition into two large groups: score I (known as risk food for cardiovascular diseases) and score II (known as healthy food). The association between the scores and serum blood lipoprotein levels among the study population was analyzed through multiple linear regression analyses. Modeling step-wise techniques were used to enter the covariates into the linear models. RESULTS: Increasing mean levels of TOTAL-C and LDL-C were seen from the lowest through the highest quintile of score I when compared to the score II, where decreasing mean levels of TOTAL-C and LDL-C were observed from the lowest to the highest quintile. The results of the linear regression analyses between serum TOTAL-C and LDL-C levels for both FFC score I and score II, after multivariate adjusting, showed a significant positive relationship with score I and a significant and inverse relationship with score II. CONCLUSIONS: In population studies, FFC analyses through scores can be the choice method to evaluate the quality of diet and its potential effect on serum levels of TOTAL-C and LDL-C. PMID- 11887225 TI - [Growth and work among elementary and high school students in Sao Paulo, Brazil]. AB - OBJECTIVE: To assess risk factors for low height and students and working adolescents in cities of State of Sao Paulo, Brazil. METHODS: A stratified sample, consisting of 50.0% of students from 5th grade to last year of high school, of State of Sao Paulo, Brazil, in 1999, was drawn from two largest schools of two different cities (urban and rural). A total of 756 individuals were studied. The height/age indicator, according to the 1977-NCHS (National Center for Health Statistics) standards, defined malnutrition. Height/age distribution and multivariate analyses were carried out using the stepwise method and low-height as the dependent variable. RESULTS: Of the adolescents, 12.7% (96) fell below percentile 5; 24.4% (184) between percentiles 5 - 15; and 47.1% (356) between percentiles 15 - 50. Low height was associated with age: taking age-group 10-13 as reference, low-height was twice as likely in students aged 14-17 years (OR adj.=2.49). For those aged 17-19 years, low height was three times as likely (OR adj.=3.37). Being unemployed increases the risk for low-height (OR adj.=2.86) when compared to working adolescents. Also, low height is higher (OR adj.=1.81) among part-time workers. CONCLUSION: Economical determinants contribute to the risks for chronic malnutrition among students, since these adolescents rely on work to live on. It is worth emphasizing that underage labor legislation should be enforced in conjunction with compensation public programs. PMID- 11887226 TI - [Magnitude, geographic distribution and trends of anemia in preschoolers, Brazil]. AB - OBJECTIVE: To assess the magnitude, geographical distribution and trends of the prevalence of nutritional anemia among preschoolers and to identify risk groups in the state of Paraiba, Brazil. METHODS: A cross-sectional survey with multistage random sampling procedure was conducted in 8 urban municipalities of three mid-regions (Zona da Mata, Agreste and Sertao) in the state of Paraiba in 1992. A total of 1,287 preschoolers of both sexes were selected. Hemoglobin was determined by the cyanometahemoglobin method in venous blood, employing <11 g/dl as the cut-off for anemia. Statistical analysis of proportions employed the chi Square test, whereas for means Mann-Whitney and Kruskal-Wallis were the choice, all with confidence interval of 95%. RESULTS: The prevalence of anemia was 36.4% (CI 33.7 - 39.1) in the state of Paraiba, greater (p=0.00) than that observed (19.3%, CI 17.3 - 21.5) in 1982. Only 1.0% (CI 0.61 - 1.8) and 6.8% (CI 5.5 - 8.3) of anemia cases were classified as severe and moderate, respectively. Boys presented lower mean hemoglobin concentration (p=0.00), and children under age 3 comprised the biological group of highest susceptibility for deficiency status (p= 0.00). It was found that the second year of life is the most critical for developing nutritional deficiency (p= 0.00). The Agreste mid-region revealed to be the geographical area of highest risk (p= 0.00), outlining a new epidemiological dynamics when compared to the year 1982, when the drought-ridden Sertao region were the geographical area at greatest risk of deficiency. CONCLUSIONS: According to international epidemiological criteria, anemia in the studied regions represents a public health problem of moderate degree Assuming the analytical comparability of the 1982 and 1992 cross-sectional surveys, it suggests an increasingly prevalence of nutritional anemia (+88.5%) in all 3 mid regions in a 10-year-period (1982-92). PMID- 11887227 TI - [Medication directions as a source of technical and scientific information]. AB - OBJECTIVE: To assess the adequacy and quality of information available in package directions of basic medicines commercialized in Brazil, according to both the specialized technical and scientific literature and the current regulations on the subject. METHODS: One hundred and sixty-eight medication directions of medicines containing forty-one specific drugs selected from the Brazilian National Inventory of Essential Medication. The analysis was carried out using as reference the Brazilian Regulation (Portaria SVS 110/97) and USP-DI Technical and Scientific Reference Literature-1999. Data was collected using an evaluation form, elaborated based on the guide for medicine directions, and each section of the directions was then analyzed according to the level of satisfaction previously established in the literature. RESULTS: The results show that 91.4% of medication directions addressed to the patient were unsatisfactory, whereas 97.0% were inappropriate regarding their technical content. Information was either incomplete or incorrect. CONCLUSION: These circumstances are very likely a result of lack of appropriate official regulations and surveillance by government agencies associated with low level of social monitoring on the consumers' side. PMID- 11887228 TI - [Prevalence and risk factors associated with drug use among school students, Brazil]. AB - OBJECTIVE: To assess prevalence and risk factors associated with drug abuse among public elementary and high school students in the southern city of Florianopolis, Brazil. METHODS: A descriptive cross-sectional study was carried out using a standardized questionnaire created during the 4th National Survey on Drug Abuse. Four hundred and seventy-eight students were interviewed by trained college students. Of the interviewees, 43% aged 13 - 15 years and 32% aged 16 - 18 years and they had a higher socioeconomic status than the national average. RESULTS: Ever use prevalence for alcohol, marijuana, solvent drugs and amphetamines was 86.8%, 19.9%, 18.2% and 8.4%, respectively. Regular use (6 or more times per month) of alcohol, marijuana, solvent drugs and amphetamines was found in 24.2%, 4.9%, 2.5% and 2.3% of students, respectively, a higher percentage when compared to other southern states' capitals and the national average. Age, sex, social status and living with both parents were significantly associated with drug abuse. Girls were twice as likely to consume weight loss drugs and stimulants, and almost three times more likely to use tranquilizers without medical prescription. Boys were almost twice as likely to use solvent drugs. Higher social students were twice as likely to consume alcohol than those of lower social status. Cigarette and marijuana smoking, respectively, were 84% and 67% more likely among students whose parents were separated. CONCLUSIONS: There is a high prevalence of drug use among elementary and high school students in Florianopolis. PMID- 11887229 TI - [Blood alcohol content prevalence among trauma patients seen at a level 1 trauma center]. AB - OBJECTIVE: To investigate the prevalence of blood alcohol content (BAC) among patients seen at a level 1 trauma center. METHODS: A cross-sectional study was carried out and patients were randomly selected at the emergency room of a level I trauma center in the city of Sao Paulo, Brazil, throughout a year (August 1998 to August 1999). Blood samples were drawn and data was collected using an adapted version of a questionnaire developed by the Medical Research Institute of San Francisco - Alcohol Research Group. RESULTS: A population sample of 464 patients was analyzed. Most of them were males (73.7%) and the median age was 29 years old. Positive BAC was found in 28.9% of the cases (CI95% 24.8 - 33.2) and in 84.3% BAC was =0.10%. Type of injury, gender, age group, marital status and outcome showed statistically significant associations with BAC with the highest BAC prevalence observed among assault victims (46.2%), males (33.9%), 25 to 44 years old (37.6%), singles (33.0%), and patients admitted in the hospital (41.4%). CONCLUSIONS: The results reinforce the relationship of alcohol and trauma. Preventive actions at different levels focusing on higher risk groups for alcohol-related injuries should be considered as part of prevention programs to both reduce injuries and curb recurrent events. PMID- 11887230 TI - [Prevalence and factors associated with self-medication: the Bambui health survey]. AB - OBJECTIVE: A population-based study was carried out in the municipality of Bambui, Brazil (population: approx. 15,000 inhabitants), to determine the prevalence of self-medication and its associated factors. METHODS: A random sample of 1,221 residents aged >18 years was selected. Of these, 796 reported use of medications in the last 90 days and were selected for this study (775 participated). Data was collected through home interviews. Study variables were divided in 3 groups: social and economic, health status and health service use indicators. Statistical analysis was performed using Pearson's Qui-square test, and odds ratios adjusted by multinomial logistic regression. RESULTS: Of the total, 419 (54.0%) reported use of only prescribed medications, 133 (17.2%) took prescribed and over-the-counter medications, and 223 (28.8%) took only over-the counter medications in the last 90 days. After adjusting for confounders, the following variables presented significant associations with exclusive use of self medication: female sex (OR=0.6; IC95%=0.4 - 0.9); age (OR=0.4; IC95%=0.3 - 0.6 for 40-59 years old and OR=0.2; IC95%=0.1 - 0.5 for >60 years); >5 residents in the household (OR=2.1; 1.1 - 4.0); number of visits to a doctor in the previous 12 months (OR=0.2; IC95%=0.1 - 0.4 and OR=0.1; IC95%=0.0-0.1 for 1 visit and >2 visits, respectively); report of consulting a pharmacist in the previous 12 months (OR=1.9; IC95%=1.1 - 3.3); and reports of financial expenses with medications during this period (OR=0.5; IC95%=0.3 - 0.8). CONCLUSIONS: The study results show that the prevalence of self-medication in the studied community was similar to that observed in developed countries. These results also suggest that self-medication works in place of the formal health attention in this community. PMID- 11887231 TI - Detection of anti-Giardia lamblia serum antibody among children of day care centers. AB - OBJECTIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA) for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3%) showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3%) and 100 (68%) samples, respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39%) than IIF (70%). IIF also showed a higher concordance with microscopic examination than ELISA. PMID- 11887232 TI - [Spatial distribution of Ascaris lumbricoides infection]. AB - OBJECTIVE: To estimate risk areas for Ascaris lumbricoides parasitic overload, using geoprocessing and geostatistic methods of analysis. METHODS: A coproparasitologic and domiciliary survey was conducted in 19 selected census districts of the state of Rio de Janeiro, Brazil. A sample of 1,664 children aged between 1 - 9 years was selected and plotted in their own home' centroid. Geostatistics techniques allowed spatial exploratory analysis, variographic study, and ordinary kriging. Student t-test, odds ratio and confidence intervals were used in the statistical analysis. RESULTS: A prevalence of 27.5% was found for A. lumbricoides. Household income, housewife's education level and peridomiciliary conditions were identified as significantly associated factors to the occurrence of ascariasis. An isotropic spherical semivariogram model with 150 m reach, contribution of 0.45 and nugget effect of 0.55 was employed in ordinary kriging. CONCLUSIONS: Peridomiciliary impact on ascariasis is confirmed by a spatial continuity of 150 m. Disease occurrence could be estimated in the study area and a risk map elaborated using ordinary kriging. PMID- 11887233 TI - Intra-population plasticity of Anopheles darlingi's (Diptera, Culicidae) biting activity patterns in the state of Amapa, Brazil. AB - OBJECTIVE: To assess the variation in Anopheles darlingi's biting activity compared to An. marajoara in the same locality and to biting activity data from other regions. METHODS: Using human bait, eight observations of the biting activity of An. darlingi and An. marajoara were carried out during 1999 and 2000 in the municipality of Sao Raimundo do Pirativa, state of Amapa, Brazil. Each observation consisted of three consecutive 13-hour collections, close to full moon. There were shifts of collectors in the observation points and nocturnal periods. RESULTS: An. darlingi revealed considerable plasticity of biting activity in contrast to An. marajoara, which showed well-defined crepuscular biting peaks. No significant correlation between density and biting activity was found, but a significant correlation existed between time and proportional crepuscular activity, indicating underlying ecological processes not yet understood. Two of the four available data sets having multiple observations at one locality showed considerable plasticity of this species' biting patterns as well. CONCLUSION: Intra-population variation of biting activity can be as significant as inter-population variation. Some implications in malaria vector control and specific studies are also discussed. PMID- 11887234 TI - Reproductive outcomes in an area adjacent to a petrochemical plant in southern Brazil. AB - OBJECTIVE: To evaluate possible adverse reproductive outcomes in an area adjacent to a petrochemical plant in southern Brazil. METHODS: A review of 17,113 birth records of the main hospital of the municipality of Montenegro, southern Brazil, from 1983 to 1998 was carried out. Three groups of cases were selected: (1) newborns with major congenital malformations; (2) newborns with low birth weight (<2,500 g); and (3) stillborns (>500 g). A control was assigned to each case. Controls were the first newborns weighing 3). Although the level of risk was similar between the two bereaved groups, it was significantly different from those in coupled relationships, who had the lowest risk (1.43). Based on the interviews, the dietary issues included: 1) food acquisition, preparation and consumption; 2) difficult meals place and time; 3) influence of social network/spouse; and 4) food and nutrition information. The food-related issues faced by bereaved individuals were similar, but substantially different from those in coupled relationships. CONCLUSIONS: The results show that bereavement counseling does not serve as a gateway to reduced nutritional risk and highlights the need to address food issues in grief resolution interventions. PMID- 11887240 TI - Longitudinal study of effects of alcohol use and/or personal exercise on high density lipoprotein cholesterol in aging humans. AB - Self-controlled alcohol use and/or personal physical exercise is generally believed to be advantageous in extending good health in aging humans. We investigated whether data collected over 31 years in the same subjects (369 men, 75 women) in our Longitudinal Aging Study of humans (1969-2000) showed strong relationship between alcohol and/or physical exercise with High Density Lipoprotein Cholesterol (HDLC). Women s HDLC interactions among age, diet, alcohol use and/or exercise were not found to be statistically significant in our study. Men, however, showed a strong positive statistical significance (p80 yr). Energy requirements should be estimated from measurements of energy expenditure. DESIGN AND METHODS: 21 free living individuals (8 males, 13 females), 91-96 years of age, living in Goteborg, Sweden were studied by the doubly labelled water method (DLW) for measuring total energy and by a ventilated hood system for Resting metabolic rate (RMR). RESULTS: RMR averaged 5.36 (SD 0.71) MJ/d in females (n=12) and 6.09 (SD 0.91) MJ/d in males (n=8). Difference between measured RMR and predicted basal metabolic rate (BMR) (n=20) was 0.015 (SD 0.86) MJ/d (NS). Total energy expenditure (TEE) measured by DLW averaged 6.3 (SD 0.81) MJ/d in females and 8.1 (SD 0.73) MJ/d in males. Activity energy expenditure (AEE=TEE-RMR), thus including diet induced thermogenesis, DIT) averaged 0.95 (SD 0.95) MJ/d in females (n=12) and 2.02 (1.13) MJ/d in males. Physical activity level (PAL=TEE/BMR) averaged 1.19 (SD 0.19) in females and 1.36 (SD 0.21)(p=0.08) in males. DISCUSSION: If DIT is assumed to be 10 per cent of TEE, energy spent on physical activity will be very low in this very old population. PMID- 11887243 TI - Cumulative effects of cardiovascular disease risk factors on quality of life. AB - OBJECTIVES: The Medical Outcomes Study Short-Form Health Survey (SF-36) was used to assess the quality of life for adults who differed in level of risk for cardiovascular disease. METHODS: Subjects were 51 men and 80 women from southwestern Ohio between the ages of 20 and 86 years. Individuals level of risk was based on the culmination of four cardiovascular disease risk factors: hypertension (i.e., systolic BP>or=140 mmHg or diastolic BP>or= 90 mmHg), obesity (i.e., BMI>or=30), high cholesterol (i.e., total cholesterol>or=240 mg/dL), and presence/absence of smoking. RESULTS: Each risk factor was analyzed independently and cumulatively for effects on the SF-36 dimensions (i.e., Physical Functioning, Role-Physical, Bodily Pain, General Health, Vitality, Social Functioning, Role Emotional, Mental Health). The data suggested that quality of life impairment (indicated by lower scores on the SF-36 dimensions) increased as the number of cardiovascular disease risk factors an individual had increased. CONCLUSIONS: Cardiovascular disease risk factors unknown to the participants had differential effects on the SF-36 dimensions, and quality of life decreased as the number of risk factors individuals had increased. PMID- 11887244 TI - Evaluation of nutritional status and its relationship with functional status in older citizens with diabetes mellitus using the mini nutritional assessment (MNA) tool--a preliminary investigation. AB - BACKGROUND: Despite all that is known about diabetes mellitus, little is known about the nutritional status of older adults with this condition. OBJECTIVE: To determine whether older people with diabetes mellitus are malnourished when compared with non-diabetic subjects, to evaluate the MNA in this group, and to assess the possible relationship between nutritional impairment and function. DESIGN: A case controlled study. Community-dwelling volunteers were selected randomly from 2 general practice registers. 35 people over the age of 65, with diabetes mellitus, were age and sex matched with 35 control subjects without diabetes. The major outcome measures were: the MNA questionnaire, anthropometric measurements, serum albumin, transferrin, Barthel Index, Nottingham Extended ADL score and handgrip. RESULTS: The diabetic group scored significantly lower on the MNA than the control group (p< 0.01), but this was mainly indicative of many of the diabetic subjects scoring within the at risk category of the tool. Those in the diabetic group also had significantly lower albumin scores (P<0.05) when compared with the control group. Within the diabetic group, and in the study group as a whole, the MNA scores were significantly correlated with Barthel Index (p<0.01), Nottingham Extended ADL score (p<0.01) and handgrip (p<0.01). CONCLUSION: Community dwelling elderly subjects with diabetes may be at risk of malnutrition when compared with non-diabetic citizens. There is probably a causal relationship between malnutrition and functional decline in this group. Further research is needed, where the prevalence of malnutrition is higher, to fully evaluate the MNA in people with diabetes. PMID- 11887245 TI - Weight loss and metabolic changes in dementia. AB - Weight loss is common in elderly people with dementia, particularly those with Alzheimer s disease, and feeding difficulties are major issues in their care in the later stages of their disease. This review summarizes data from cross sectional and longitudinal studies of weight changes with dementia, physiologic aspects of the metabolic and nutritional changes in dementia, and treatment strategies to minimize weight loss. PMID- 11887246 TI - A review of the state of research into the role of iron in stroke. AB - Iron is a double edged sword for living systems, as it is essential for a wide range of metabolic processes while it also has potential deletereous effects. Oxidative metabolism during ischaemic stroke together with high iron content in the brain synergise to increase the oxidative damage. High plasma ferritin, as a measurement of iron stores, and high cerebrospinal fluid ferritin have been related to poor outcome in stroke patients. Iron has been found in pooled gruels of atheromatous lesions and has been related to other diseases. Further epidemiological studies are required to determine the effect of iron on the development of cardiovascular diseases. Until the precise effect of iron overloading is established it is recommended that iron supplements should only be prescribed when there is a clear deficiency. PMID- 11887247 TI - Dietary antioxidants, peroxidation and cardiovascular risks. AB - Most of the many epidemiological studies in the field strongly suggest that an equilibrated diet such as the so-called "mediterranean diet", is associated with protective effects against major diseases, and particularly, against cardiovascular risks. Since many reports also consider reactive oxygen species or free radical oxidations to be responsible for the accompanying disorders of most pathologies as well as for ageing, it is conceivable that natural plant metabolites such as polyphenols, are likely to play an important role in insuring this protection. Indeed, not only their presence, in particularly high amounts and varieties in foods of such a diet, but also, inter alia, their very potent antioxidant or radical scavenging properties, make polyphenols best accounting for the parodoxical part of the french paradox . Therefore, many efforts have been made to assess the mechanisms for such a cardiovascular disease protection. Whatever convincing were the polyphenols properties demonstrated by many in vitro experiments to support those theories, quite a great number of the results appeared somewhat contradictory when transposed to humans, in the in vivo situation. Some people totally refute this explanation, thinking that health benefits, as far as alcoholic beverages are concerned, originate from ethanol but also, with no doubt, some polyphenols even revealing to be pro-oxidants . PMID- 11887248 TI - [Dopplersonography of the ductus venosus: assessment, evaluation and actual clinical importance]. AB - The ductus venosus (DV) connects the intra-abdominal umbilical vein with the infundibulum of the IVC and develops during pregnancy to a trumpet-shaped structure with a narrow isthmus that accelerates the blood jet crossing the IVC directly to the left atrium via the foramen ovale avoiding mixing with deoxygenated blood from the right chamber. In animal studies, blood flow and doppler sonographically analyzed blood flow velocity waveforms mainly is controlled by heart rate and central venous pressure. The velocity waveform of the DV contains two peak components: the first indicates systolic velocity of the ventricle, the second peak diastolic velocity. A nadir is seen during atrial contraction. In animal studies, DV blood velocity in hypoxemia is influenced by central venous pressure and heart rate. The determination of the DV/UV ratio reflects the redistribution of blood flow, increases in hypoxemia and is therefore more reliable than blood velocity measurement for the detection and evaluation of fetal distress. In cases with severely growth-restricted fetuses, recipient twin in TTTS (twin-to-twin transfusion syndrome), tachyarrhythmia induced cardiomyopathia and congenital heart disease, the measurement and interpretation of DV Doppler waveform pulsatility seems to be a useful tool that provides important information on the fetal condition and outcome. In cases of zero or reverse flow during atrial contraction in most cases the delivery of the fetus is indicated. An improvement of morbidity and mortality using Doppler sonography of the DV has not yet been proven. In cases of fetuses with or without chromosomal aberrations with major defects of the heart it can be used in addition to the standard screening methods of the first trimester of pregnancy for detection of heart failure. PMID- 11887249 TI - [Results of a National Survey in Germany on incidence and therapy of the nonoliguric hyperkalemia of the premature infant]. AB - BACKGROUND: Although nonoliguric hyperkalemia of the premature infant is a common problem, in many hospitals it is not observed. It is characterized by an excessive increase in serum potassium concentration (serum-[K (+)]) at 24 hours after birth. Recently, salbutamol has been recommended to treat hyperkalemia. There are no controlled trials in premature infants. During the first minutes of salbutamol infusions paradoxical increases in serum-[K (+)] may occur. We asked for possible reasons for the variable incidence of the disorder. We wanted to know if salbutamol is currently applied and if initial increases in serum-[K (+)] during salbutamol infusions have been observed. MATERIALS AND METHODS: National survey in Germany. RESULTS: Questionnaires of 132 hospitals caring for premature infants < 30 gestational weeks. The incidence of hyperkalemia was 0 - 40 %. At least 25 % of all hospitals measured serum-[K (+)] not before the second day after birth. 52 hospitals had applied salbutamol. Nine hospitals reported increases in serum-[K (+)] associated with salbutamol therapy. However, it was common practice to measure serum-[K (+)] 1 hour (median, range 0.25 - 12 hours) after the start of the infusion. Two cases of cardiac arrhythmias during salbutamol infusion were reported. DISCUSSION AND CONCLUSIONS: The variable incidence of hyperkalemia may result from the fact that many hospitals do not measure serum-[K (+)] on the first day after birth. Common practice made it potentially impossible to determine the frequency of initial increases in serum [K (+)] during salbutamol infusion. However, these data should be available, before controlled trials on salbutamol treatment in premature infants can be initiated. PMID- 11887250 TI - [Lavage with exogenous surfactant in neonatal meconium aspiration syndrome]. AB - BACKGROUND: Meconium aspiration syndrome is a disease in near term infants which requires invasive techniques to decrease mortality. Lavage of the tracheobronchial tree with exogenous surfactant has been shown to be effective in animal studies. We studied the effectiveness of this technique in newborn infants and compared them with a historical control group. PATIENTS AND METHODS: From 1987 to 1998, we treated 18 neonates with meconium aspiration syndrome. In 11 babies a lavage with bovine derived diluted surfactant was carried out immediately after admission. In 7 infants of the control group only the meconium was suctioned or a lavage with saline performed. There were no differences between both groups concerning other therapeutic interventions like high frequency ventilation or inhalative NO. The control group contained significantly more outborn patients compared with the lavage group. Criteria of effectiveness were change in oxygenation index (OI), duration of mechanical ventilation and oxygen supplementation greater than 30 %. In addition, the two groups were compared for incidence of death, need for ECMO, and air leaks. RESULTS: One infant of the lavage group died, one had to be transferred to ECMO (no significant difference). There was a significant decrease in OI after surfactant lavage from 22 at admission to 5.1 one day later (p=0.007), whereas in the control group, OI did not change significantly over time (15.8 to 11.4). There were no differences in the duration of mechanical ventilation or oxygen supplementation. CONCLUSIONS: In our study, lavage with exogenous surfactant had only a short-term effect in decreasing OI in neonates with meconium aspiration. Larger numbers of patients need to be investigated to demonstrate an improvement in long-term outcome. PMID- 11887251 TI - [Prenatal manifestation of a congenital glioblastoma - Case report]. AB - In prenatal ultrasound screening, internal hydrocephalus and intracranial bleeding of the fetus are considered as primary diagnostic signs for a congenital brain tumor. We report the prenatal sonographic diagnosis of a congenital glioblastoma due to acute fetal internal hydrocephalus in the 37th week of gestation. After birth, the tumor's hyperechoic appearance on ultrasound was indistinguishable from intracranial bleeding. Diagnosis of a congenital glioblastoma (WHO stage IV) was confirmed by subtotal tumorectomy in the 9th week of life. In the international literature, only 6 cases of prenatally diagnosed glioblastomas have so far been reported, all of which associated with sonographically diagnosed fetal hydrocephalus. Further sonographic signs for a brain tumor are the tumor mass itself, a polyhydramnion, enlarged biparietal diameters and head circumferences, as well as suspected intracranial bleeding. PMID- 11887253 TI - [Neonatal alloimmune thrombocytopenia with hemorrhagic infarction of the right hemisphere]. AB - Alloimmune thrombocytopenia is the most common cause of severe thrombocytopenia (< 20 000/microliter) in otherwise healthy neonates. The diagnosis is usually made in the diagnostic workup of neonatal hemorrhage. If diagnosis and therapy are delayed the consequences for the affected child can be deleterious. Case report of a full-term neonate with the clinical symptoms of severe hemorrhage and hemorrhagic infarction of the right hemisphere due to neonatal alloimmune thrombocytopenia. While the plasmatic coagulation parameters were unaffected, we detected severe neonatal thrombocytopenia and anemia and identified alloimmune antibodies against the child's thrombocytes in the mother's serum. The antibodies were specific against the platelet antigen HPA-1a. Transfusion of HPA-1a-negative thrombocytes stabilized the platelet count. Anemia was treated by erythrocyte transfusion. Unfortunately, the patient exhibited, most likely intrauterine, intracranial hemorrhage and infarction of the right hemisphere, the most dreaded complication of neonatal alloimmune thrombocytopenia. The identification of severe thrombocytopenia causing the hemorrhage allowed us to start substituting thrombocytes without any delay. The previous diagnosis of alloimmune thrombocytopenia should lead to monitoring a subsequent pregnancy in a specialized unit allowing fetal blood sampling and intrauterine thrombocyte substitution if necessary.This case report exemplifies the symptoms and treatment of neonatal alloimmune thrombocytopenia. The differential diagnosis for a neonate showing clinical signs of hemorrhage should include alloimmune thrombocytopenia which can then be treated adequately. The diagnosis should also lead to careful monitoring of subsequent pregnancies. PMID- 11887252 TI - [Intrauterine therapy and outcome in four pregnancies of one mother with anti ro autoantibody positive Sjoegren's syndrome]. AB - Autoantibodies against 52/60kD-Ro proteins, frequently present in patients with Sjoegren's Syndrome or systemic lupus erythematosus, are transmitted to the fetus during pregnancy. These autoantibodies can damage the cardiac conductive system of the fetus and cause a complete atrioventricular block, with a mortality of 30 %. We report the intrauterine therapy during four pregnancies of the same mother with high 52/60kD-Ro autoantibodies and the outcome of her infants. Our patient with primary Sjoegren's Syndrome suffered an early miscarriage during her first pregnancy. During the second pregnancy, a fetal atrioventricular block was observed at 23 weeks of gestation. Although subsequently dexamethasone therapy and daily plasmaphereses were started, a cesarean section was necessary at 26 weeks due to hydrops fetalis. The girl died from the atrioventricular block after two days. During the third and fourth pregnancies, dexamethasone therapy was begun already at 7 weeks, and regular plasmaphereses at 15 weeks. The children were delivered by cesarean section at 32 and 36 weeks because of growth retardation. Both had normal electrocardiograms after birth and after 2 and 4 years. In pregnant women with connective tissue diseases, monitoring of anti Ro autoantibodies and fetal heart function is important. Intrauterine therapeutic options are dexamethasone therapy to suppress maternal and fetal inflammatory reactions and repeated plasmaphereses to reduce autoantibody levels. Postnatal follow up of the infants for atrioventricular block and rheumatic manifestations is necessary. PMID- 11887254 TI - Scoping practice issues in the Australian mental health nursing workforce. AB - This is the third of four articles on the scoping study of the Australian mental health nursing workforce conducted on behalf of the Australian and New Zealand College of Mental Health Nurses (ANZCMHN) for the Australian Health Ministers Advisory Council (AHMAC) National Working Group on Mental Health (NWGMH). Its purpose is to focus on factors that significantly affect mental health nursing practice. The issues of advanced practice, regulation of nursing, accreditation, credentialing and demarcation with other disciplines are addressed. PMID- 11887255 TI - The meaning and significance of clinical group supervision and supervised individually planned nursing care as narrated by nurses on a general team psychiatric ward. AB - By interviewing 22 psychiatric nurses, the present study aimed to reveal the meaning and significance of systematic clinical group supervision and supervised individually planned nursing care, using latent content analysis. The interpreted meaning was 'confronting the complexity of ongoing life in daily nursing care' and the interpreted significance was 'strengthening the foundation for nursing care'. Reflection on action and confirmation seemed to be core components in the process of clinical supervision. Focusing on the relational and task aspects in nursing care within a group approach may have contributed to the positive experiences of development that occurred. PMID- 11887256 TI - 'Psychiatric nursing was great, but I want to be a "real" nurse': is psychiatric nursing a realistic choice for nursing students? AB - The present paper reports the results of a quasi-experimental study that examined the relationship between exposure to the theory and practice of psychiatric nursing and the desirability of psychiatric nursing as a future career choice. A time series design was utilized to enable a comparison between pre- and post-test scores. A significant increase in the popularity of psychiatric nursing was evident in the experimental group in the post-test phase, while no significant change was detected in the control group. Despite this increase, a large number of students from the experimental group indicated their reluctance to undertake a career in psychiatric nursing without first consolidating their skills in the medical-surgical area. This situation reflects an image of nursing that is contrary to the caring philosophy it purports to embrace, but which appears to favour certain areas of nursing practice at the expense of others. PMID- 11887257 TI - Are universities preparing nurses to meet the challenges posed by the Australian mental health care system? AB - The preparedness of comprehensive nurses to work with the mentally ill is of concern to many mental health professionals. Discussion as to whether current undergraduate nursing programs in Australia prepare a graduate to work as a beginning practitioner in the mental health area has been the centre of debate for most of the 1990s. This, along with the apparent lack of interest and motivation of these nurses to work in the mental health area following graduation, remains a major problem for mental health care providers. With one in five Australians now experiencing the burden of a major mental illness, the preparation of a nurse who is competent to work with the mentally ill would appear to be a priority. The purpose of the present study was to determine third year undergraduate nursing students' perceived level of preparedness to work with mentally ill clients. The results suggested significant differences in students' perceived level of confidence, knowledge and skills prior to and following theoretical and clinical exposure to the mental health area. Pre-testing of students before entering their third year indicated that the philosophy of comprehensive nursing: integration, although aspired to in principle, does not appear to occur in reality. PMID- 11887258 TI - Does drawing up technique influence patients' perception of pain at the injection site? AB - Traditionally nurses have used one of two drawing up techniques for the administration of intramuscular (i.m.) injections. In the first, the injectable is drawn up using one needle, which is then discarded before administration using a new needle. Alternatively, the injectable is drawn up and administered without changing the needle. Advocates of the two-needle technique suggest that this method reduces pain at the injection site. In the present study, 70 subjects completed an independently validated pain scale following administration of an i.m. depot neuroleptic. The conclusion, that there is a significant reduction in injection site pain, using the two-needle technique, is not supported by the data obtained in this study. PMID- 11887259 TI - Refocus mental health nursing. PMID- 11887260 TI - Demystifying healthcare corporate compliance programs. PMID- 11887261 TI - California law regarding living wills. PMID- 11887262 TI - Our emergency department is structured without "walls" and we have curtains that separate many of the patient care areas. PMID- 11887263 TI - A guide to conversion foundations transition. PMID- 11887264 TI - Domestic violence. AB - Domestic violence, a serious health problem, affects millions of people every year. Healthcare providers are in a unique position to identify and provide intervention in such cases. Unfortunately, providers under diagnose victims of domestic violence. Thus, some states have mandated reporting of domestic violence. This article presents the advantages and disadvantages of mandatory reporting and suggests alternatives that improve the response to domestic violence in the healthcare setting. PMID- 11887265 TI - United Kingdom (UK) post-liberal laws/policies and the mentally disordered. PMID- 11887266 TI - Scoping the prospects of Australian mental health nursing. AB - In March 2000 the Australian & New Zealand College of Mental Health Nurses submitted the final report on the National Scoping Study of Mental Health Nursing in Australia to the Mental Health Branch of the Department of Health and Aged Care. In this final article, in a series of four, the authors present an overview of the future prospects of mental health nursing in Australia. PMID- 11887267 TI - The use of the Liverpool University Neuroleptic Side-Effect Rating Scale (LUNSERS) in clinical practice. AB - Forty-four mental health clients completed the Liverpool University Neuroleptic Side-Effect Rating Scale (LUNSERS)--a self-rating scale to assess the prevalence and intensity of neuroleptic side-effects. In the month prior to the study, 50% of the clients surveyed had experienced more than half of the side-effects outlined on the 41-item scale. A prevalence profile allowed us to rank the frequency of individual side-effects across the sample. Some side-effects such as 'difficulty concentrating', 'difficulty remembering', 'tiredness' and 'restlessness' were experienced by most of the clients in the study while 'unusual skin marks', 'difficulty passing water', 'rashes' were experienced by a few. A prevalence profile may be a useful guide in developing strategies for managing side-effects more effectively in small groups of clients. In addition, the use of the LUNSERS in clinical practice would enable case managers to establish baseline measures for individual clients and evaluate changes in medication and other non-medical strategies for reducing unwanted side-effects. The identification and assessment of antipsychotic side-effects is an important area for client and professional carer education. PMID- 11887268 TI - Identification and monitoring of disordered water balance: bioelectrical impedance analysis as an alternative to the target weight procedure. AB - Many psychiatric patients present with disorders of water balance manifested by polydipsia or polyuria. Multiple frequency bioelectrical impedance analysis (MFBIA) was investigated as a suitable method for the assessment of body water in such patients. Whole body bioelectrical impedance, total body water and extracellular water and serum sodium were measured in 10 chronic psychiatric inpatients with polydipsia. Fluid volumes predicted by MFBIA correlated significantly (P < 0.002) with those obtained by reference methods but the relatively high limits of agreement (2.2 L) suggest that the method is not yet sufficiently accurate for random screening purposes. However, owing to the simplicity and non-invasive nature of the technique, it warrants further investigation and development, particularly for routine patient monitoring. PMID- 11887269 TI - Nurse--client relationships: the experience of community psychiatric nurses. AB - The aim of the present study was to construct an interpretation of the experience of the nurse--client relationship in the context of community psychiatric nursing. Hermeneutic phenomenology formed the framework of the study. Shared conversations were conducted with five experienced community psychiatric nurses and five clients. Themes of 'Being there', 'Being concerned', 'Establishing trust' and 'Facilitating transition' were identified from the nurses' conversations. This thematic structure was used to illuminate the centrality of the nurse--client relationship, and to articulate the skills that are involved in establishing and maintaining the relationship with clients with mental illness. PMID- 11887270 TI - An exploratory investigation into the housing preferences of consumers of mental health services. AB - In the present qualitative study, 10 mental health consumers living in the community were interviewed in relation to their housing situations. Three themes emerged from the interview data: 'The place', 'Other consumers' and 'A normal life'. Consumers rejected congregated housing because it enables the larger community to more easily identify them as former psychiatric patients. The views of participants consistently indicate a desire to live a normal life. Results of this study highlight the need for greater consumer participation with mental health nurses and policy makers, in relation to the development of housing services. PMID- 11887271 TI - $75 million of stuff. PMID- 11887272 TI - Giving lessons in love. PMID- 11887273 TI - Identifying inappropriate sinus tachycardia. AB - This rare arrhythmia is often misdiagnosed. Learn to recognize it and give patients the treatment they need. PMID- 11887275 TI - End points of resuscitation: choosing the right parameters to monitor. AB - Determining when resuscitation is complete can be challenging, as tissue hypoperfusion can persist despite normal vital signs. This article discusses the limitations of traditional parameters used as resuscitation guidelines and describes new technologies that aid in assessing resuscitation efforts, including advances in hemodynamic monitoring and methods for obtaining global and organ specific indexes. PMID- 11887274 TI - How to recognize and treat propellant inhalation. AB - Cheap and abundant, inhalants are an easy way for young people to get high. These substances are physically and psychologically addicting, and can cause death. Find out what nurses--and parents--need to know. PMID- 11887276 TI - Post myocardial infarction treatment in the older adult. AB - Treatments for acute myocardial infarctions (AMIs) have advanced over the past few decades. Although AMIs are considered medical emergencies, continuing research has provided protocols and guidelines that significantly decrease mortality and reinfarction rates. Beta-blockers and aspirin are considered standard treatment for post-AMI patients; however, studies involving the elderly reveal that this population is less likely to receive beta-blocker and aspirin therapy. This article discusses current recommendations and treatments for post AMI elderly patients. PMID- 11887278 TI - Making outstandingly good presentations. AB - Try these tips for sharing information clearly while creating a positive image for yourself and for nursing. PMID- 11887277 TI - Recognizing sport diving injuries. AB - Even if scuba diving is not a local enthusiasm, someone with life-threatening dive-related problems could turn up in the emergency department at any time. This article describes how to respond. PMID- 11887279 TI - Nature's nurse: promoting sleep in the ICU. AB - This article describes nursing interventions to help patients get the rest they need to heal. PMID- 11887280 TI - Discovering meaning and purpose during recovery from an acute myocardial infarction. AB - Spirituality can play a powerful role in recovery. This article explores spirituality in patients recovering from acute myocardial infarction (AMI). By understanding the influence of spirituality on recovery, nurses can help patients discover meaning and purpose in life after AMI and be a positive influence on patient recovery. PMID- 11887281 TI - A symphony of frogs at night. PMID- 11887282 TI - The "thief of life". PMID- 11887283 TI - Organ donation. Current trends in liver transplantation. AB - Liver transplant recipients already number in the thousands in Canada. As Hepatitis C reaches epidemic proportions in North America, that number is going to increase exponentially. If liver transplantation is not affecting your practice now, it soon will be. PMID- 11887284 TI - Reality check. Do conceptual models of nursing work today? PMID- 11887285 TI - Pharmacodynamics. Quarterly medication reviews in long-term care. PMID- 11887286 TI - Client restraints more than a safety issue. PMID- 11887287 TI - Lead exposure in children. PMID- 11887288 TI - Does an employer have a right to mandate human immunodeficiency virus (HIV) testing of an employee when there is reason to believe the employee may be infected? PMID- 11887289 TI - Healthcare on-line. PMID- 11887290 TI - We'll take it! Should we? PMID- 11887291 TI - Hospital conversion foundations. Issues in creation, operation, and evaluation. AB - A major healthcare transformation in the United States is the conversion of nonprofit hospitals to for-profit entities, and the creation of hospital conversion foundations for the nonprofit charitable assets, which now exceeds $9 billion. Because less than 21% of the 525 hospitals converting from nonprofit to for-profit ownership have established a hospital conversion foundation, the public's monetary losses are considerable. This article examines some of the key legal and organizational issues related to hospital conversion foundations including factors related to establishing fair value of the converting hospital, foundation mission, use of conversion revenue, governance, and evaluation. PMID- 11887292 TI - The American Board of Nursing Specialties. Nursing's gold standard. PMID- 11887293 TI - Can a physician legally refuse to continue to provide care for a patient when the patient has been noncompliant to treatment? PMID- 11887294 TI - In cases of malpractice, are nurse practitioners held to the medical standard of care when prescribing medications for clients, or would they be held to a standard of care applicable to advanced practice nurses? PMID- 11887295 TI - Is it permissible for an organization to allow employees of a payer access to patient's medical records? PMID- 11887296 TI - Cirrhosis of liver with ascites treatment based on principles of ayurved. AB - A case of ascites with cirrhosis of liver due to chronic malaria and nutritional deficiency was treated with 20 ml of imferron with improvement and is alive for a period of six years after first treatment with iron. PMID- 11887297 TI - Antipsychotic induced movement disorders. PMID- 11887298 TI - Posterior bridging of the atlas vertebra in south Indians. AB - The vertebral artery is vulnerable to compression in its course between foramen transversarium and the foramen magnum during extreme rotation of the head and neck. This situation may be aggravated by the presence of posterior or lateral bridge of the atlas and result in compromised blood flow. The incidence of the bony ring formed by posterior bridging has been demonstrated in atlases of various races across the world: it varies between 1.875% to 29.2%. In an examination of sixty south Indian atlases it was found in 11.7% of the cases. The presence of this bony bridging should be taken in to account during a surgical manipulation of the cervical spine. PMID- 11887300 TI - Early diagnosis of neonatal sepsis using a hematological scoring system. AB - OBJECTIVE: To assess the utility of the hematologic scoring system (HSS) of Rodwell et al for the early detection of neonatal sepsis. DESIGN: Analysis of the peripheral smear findings according to the HSS by a pathologist blinded to the infection status of the neonate. SUBJECTS: One hundred and three high risk neonates having predisposing perinatal factors or clinical suspicion of sepsis. RESULTS: Analysis of the hematologic profiles in the light of the HSS found that an abnormal immature to total neutrophil (1:T) ratio followed by an abnormal immature to mature neutrophil (1:M) ratio were the most sensitive indicators in identifying infants with sepsis. These two criteria along with thrombocytopenia (< 1,50,000/cm3) had a high negative predictive value over 94%. The study also found that the higher the score the greater the certainty of sepsis being present. CONCLUSION: The HSS is simple, quick, cost effective and readily available tool in the early-diagnosis of neonatal sepsis and could provide a guideline to decisions regarding antibiotic therapy. PMID- 11887299 TI - Effect of various antihypertensive drugs on plasma fibrinogen levels in patients with essential hypertension. AB - In recent years, substantial evidence has accumulated to unambiguously implicate high plasma fibrinogen levels as a major cardiovascular risk factor. An open prospective and randomised pilot study was therefore undertaken in mild to moderate hypertensives to evaluate the effect of various antihypertensive drugs viz enalapril, felodipine and prazosin on the blood pressure and plasma fibrinogen levels. The systolic and diastolic blood pressures were determined at 0, 4 and 8 weeks whereas plasma fibrinogen assays were done at baseline and at the end of the 8th week of treatment in all the drug-treated groups. It was observed that although all the three drugs effectively controlled blood pressure, only enalapril significantly reduced plasma fibrinogen levels. Due to this additional effect, enalapril has potential to control two major cardiovascular risk factors--hypertension and high plasma fibrinogen levels--simultaneously. PMID- 11887301 TI - Thyroid hormones in treatment of mood disorders. AB - Although lithium carbonate is still the best validated and most preferred drug used for augmenting treatment of depression, T3 is perhaps the next best agent in this regard. Equivocal evidence, clinical skepticism and lack of data with SSRIs have prevented its widespread use. On the basis of whatever evidence is available it appears to be safe and effective. T4 is useful for replacement purpose and in high doses may have a role in augmenting mood stabilizers. Evidence for other therapeutic effects of thyroid hormones seems to be gathering, but they cannot be recommended yet in other situations as a part of routine clinical practice. Further research about the various therapeutic aspects of thyroid hormones is still required, since this is an area that seems to hold much promise. PMID- 11887302 TI - WHO calls on private sector to provide affordable hearing aids in developing world. PMID- 11887303 TI - Removal of organic contaminants from water or wastewater with liquefied gases. AB - This study utilized liquefied gases (LG) as extractant to remove various organic contaminants including halogenated hydrocarbons and phenols as well as aromatic compounds from aqueous matrices. Orthogonal experiments were performed to optimize the operating conditions such as temperature, co-solvents and so on. Under favorable conditions, high removal efficiencies can be readily achieved for a great number of representative model organic contaminants, the removal efficiencies for most of the hydrophobic contaminants were greater than 90% in a single extraction stage. Tentative effort was also done for the removal of extracted contaminants from recycled liquefied gases. PMID- 11887304 TI - Marine oil spill contingency planning. AB - According to the practice researching and formulating "The Oil Spill Contingency Plan of South Chinese Sea", this paper analyses and discusses the structure, functions and main contents of marine oil spill contingency planning, programs the organizing and commanding system and emergency response system, and advances the planning and researching method to coordinate comprehensively and to design practically the detailed emergency response steps until to formulate the ease operating programs for the plan implementation(PPI) and the PPI to apply high techniques supporting emergency administrations and response. PMID- 11887305 TI - Biosorption of lead by Phanerochaete chrysosporium in the form of pellets. AB - The growth of Phanerochaete chrysosporium(ATCC 24725) in pellets was influenced by culture time, medium pH, C/N, surfactant concentration, spore number in inoculum, and shaking rate. The removal of Pb2+ from aqueous solution by this kind of mycelial pellets was studied. The results indicated that many factors affected biosorption. These factors included pH, Pb2+ concentration, co-ion, adsorption time, and chemical pretreatments of biomass. Under optimum biosorption conditions(pH 4.5, 27 degrees C, 16 h), the highest lead uptake of 108 mg/g, was observed with mycelial pellets of 1.5-1.7 mm in diameter which were treated with 0.1 mol/L NaOH solution before adsorption. Pretreatment of biomass with NaOH further increased its biosorption capacity. PMID- 11887306 TI - Effects of red-yellow soil acidification on seed germination of Chinese pine. AB - Acid treatments significantly change the physical and chemical properties of red yellow soil by lowering its pH value and leaching out aluminum(Al) ions that are harmful to the growth of plants. The structure of soil will be damaged, resulting in higher viscosity, higher water retention rate and lower air permeability of the soil. The germination rate of Chinese pine(Pinus tobulacformic Carr.) seeds sowed in soil treated with sulphuric acid(H2SO4) decreased compared to that for untreated soil. The direct cause was the large amount of Al ions leached out because of low pH values(> or = 3.5). The added acid decreased the soil aggregation and increased the number of micro-aggregates(under 250 microns in diameter). Such changes increased the soil's viscosity, which tied the pine needles to the soil after the seeds had germinated and prevented the seedlings from fully developing. PMID- 11887307 TI - Chemical behavior of organic compounds in the interface of water/dual-cation organobentonite. AB - The sorption behavior of polar or ionizable organic compounds, such as p nitrophenol, phenol and aniline, in the water/organobentonite systems is investigated. Both adsorption and partition occur to the sorption of organic compounds to dual-cation organobentonites. The separate contributions of adsorption and partition to the total sorption of organic compounds to dual cation organobentonites are analyzed mathematically in the first time. The factors to the contributions are also discussed. The results indicated that the contribution of adsorption and partition is related to the composition and ratio of dual-cation surfactants exchanging onto the bentonite. The sorption of organic compounds to dual-cation organobentonite is dominated by adsorption at low concentrations and by partition at high concentrations, making the organobentonites powerful sorbents for organic contaminants over wide range of concentrations. PMID- 11887308 TI - Analysis on the environmental problems accompanying the high-speed urbanization of small towns in China. AB - The environmental problems accompanying the rapid urbanization of many small towns like Keqiao, Shaoxing County of China and their countermeasures are presented. To settle these problems rationally as well as effectively will not only conduce to boost the environmental quality and life quality of cities, but also help to promote the development of regional economy and the realization of sustainable development. PMID- 11887309 TI - An approach to the fluctuation mechanism of ecotone. AB - In this paper, the relationship among land productivity, population pressure and the fluctuation mechanism of ecotone is analyzed, taking counties and banners of southeast Inner Mongolia plateau as an example, which is the most fragile part in the farming and husbandry interleaving belt of north China due to its severe desertification degree and low level of economic development. The Cv for the total output value of agriculture decreases from southeast to northwest, with the same rule as which the overloading population cumulated distributes, and both indicators have close relation with the high linear coefficient of 0.83. These reveal the fluctuation mechanism for ecotone: fluctuation of the level of economic development is a scientific and practical measure both to weakness degree and to instability of ecotone, because it is a synthesized response to the variation of climate as well as irrational land uses, which reinforce and magnify the fluctuation. In detail, the heavier the population overloading, the severer the grassland reclamation, the stronger the dependence of regional productivity on rainfall, the lower the level of economic development, the rougher the fluctuation of ecotone, but the weaker the PRED system. PMID- 11887310 TI - Mechanisms of granular activated carbon anaerobic fluidized-bed process for treating phenols wastewater. AB - Granular activated carbon (GAC) anaerobic fluidized-bed reactor was applied to treating phenols wastewater. When influent phenol concentration was 1000 mg/L, volume loadings of phenol and CODCr were 0.39 kg/(m3.d) and 0.98 kg/(m3.d), their removal rates were 99.9% and 96.4% respectively. From analyzing above results, the main mechanisms of the process are that through fluidizing GAC, its adsorption is combined with biodegradation, both activities are brought into full play, and phenol in wastewater is effectively decomposed. Meanwhile problems concerning gas-liquid separation and medium plugging are well solved. PMID- 11887311 TI - Ecological toxicity of reactive X-3B red dye and cadmium acting on wheat (Triticum aestivum). AB - Ecological toxicity of reactive X-3B red dye and cadmium in both their single form and their combined form on wheat was studied using the experimental method of seed and root exposure. The single-factor exposure indicated that the inhibitory rate of wheat root elongation was significantly increased with the increase in the concentration of the dye in the cultural solution, although seed germination of wheat was not sensitive to the dye. The toxicity of cadmium was greatly higher than that of the dye, but low concentration cadmium (< 40 mg/L) could promote the germination of wheat seed. Interactive effects of the dye and cadmium on wheat were complicated. There was no significant correlation between the inhibitory rate of seed germination and the concentrations of the dye and cadmium. Low concentration cadmium could strengthen the toxicity of the dye acting on root elongation. On the contrary, high concentration cadmium could weaken the toxicity of the dye acting on root elongation. PMID- 11887312 TI - Optimization of C/N ratio preparation of protein-rich and multi-enzymes feed thallus through synergic fermentation of mixed distillers' grains. AB - A new procedure of determining optimal C/N (the rate of carbon source to nitrogen source) of mixed distillers' grains for combined bacteria synergic fermentation is established. At the same time an improved method evaluating bacteria growth, called method of dry cell weighing by filtering is developed. For each combination of C and N, their initial and residual contents before and after fermentation respectively are determined. Then followed the calculation of utilization of C and N sources by the compound bacteria. The optimal C/N is finally located from among the utilization of C and N of several combinations and the weight of produced mass of oven-dried thallus. The conditions of fermentation are: inoculum size 10%, temperature 30.0 degrees C, rotational speed 170 r/min, shake culture time 48 h. The best results obtained from orthogonal experiments are: maximum mass of oven dried thallus is 14.693 g in a liter liquid medium, maximum utilization rate of carbon source is 98.13% and maximum utilization rate of nitrogen is 78.14%. Optimal C/N is 5.1. PMID- 11887313 TI - Intra-specific variations of two Leymus chinensis divergence populations in Songnen Plain, northeast China. AB - Population demography, seed production, biomass allocation, net photosynthesis and transpiration of two Leymus chinensis divergent populations and between two years in Songnen plain, northeast China were compared. Strong differences between the dry 1997 and moist 1998 occurred in vegetative shoot and sexual shoot densities, sexual differentiation and tiller densities, as well as in the lengths of inflorescence, seed numbers per inflorescence, seed weights and biomass allocation in each population respectively (P < 0.01). While strong differences between the two populations occurred in vegetative shoot densities, sexual shoot densities, sexual differentiation and seed weights in each year (P < 0.01). The differences between the two populations in tiller densities and in biomass allocation to sexual shoots were significant (P < 0.05). But there were no significant differences between the two populations in the lengths of inflorescence, seed numbers per inflorescence and biomass allocation to rhizomes and vegetative shoots (P > 0.05). Excepting the transpiration rate in the early June, the differences between the two populations in net photosynthesis and transpiration rate of vegetative shoots and sexual shoots were strongly significant in the early June and July respectively (P < 0.01). Relative stable variations in population demography and physiological traits between the two populations indicated that they are divergently in the Songnen Plain. PMID- 11887314 TI - Greenhouse gas emissions from a constructed wetland for municipal sewage treatment. AB - The fluxes of greenhouse gases (methane and nitrous oxide) emission from a constructed wetland in the Eastern China as municipal sewage treatment were measured from June 1999 to August 2000 by the closed chamber method. The constructed wetland for municipal sewage treatment is a significant source of methane, up to 976.6 x 10(6) g CH4/a, which was emitted from the constructed wetland with the area of 495,000 m2 and wastewater loading rate of 12,000 m3/d. Its daily mean methane flux reached 5.22 g CH4/(m2.d), 250 times as much as that in natural wetland in the same latitude region. 227.8 mg CH4 was produced from the treatment of 1 liter wastewater, up to 700-1000 times as much as that in the secondary treatment. The emission of nitrous oxide from the constructed wetland is not higher than that from secondary treatment of wastewater, only 0.07 mg N2O/L. PMID- 11887315 TI - Cobalt in alluvial Egyptian soils as affected by industrial activities. AB - Twenty-five surface (0-20 cm) soil samples were collected from different locations in Egypt representing non-polluted, moderately and highly polluted soils. The aim of this study was to evaluate total Co content in alluvial soils of Delta in Egypt using the delayed neturen activation analysis technique (DNAA). The two prominent gamma ray lines at 1173.2 and 1332.5 keV was efficiently used for 60Co determination. Co content in non-polluted soil samples ranged between 13.12 to 23.20 ppm Co with an average of 18.16 +/- 4.38 ppm. Cobalt content in moderately polluted soils ranged between 26.5 to 30.00 ppm with an average of 28.3 +/- 1.3 ppm. The highest Co levels (ranged from 36 to 64.69 ppm with an average of 51.9 +/- 9.5); were observed in soil samples collected from, either highly polluted agricultural soils due to prolonged irrigation with industrial wastewater or surface soil samples from industrial sites. PMID- 11887316 TI - Seed yeast cultivation for salad oil manufacturing wastewater treatment. AB - The mixture of five yeast strains obtained from soil could remove about 85% TOC of oil-rich wastewater in batch test. While the highest MLSS was obtained at an N:C of 1:5, the oil removal decreased with the increase of N:C during yeast sludge cultivation. Ammonium chloride was the best nitrogen source for yeast cultivation from the viewpoint of yeast growth and oil utilization. An ammonia concentration of over 1300 mg/L led to mass death of yeast at a pH of 5. The ammonia concentration should be controlled at a level of 1000 mg/L or lower. PMID- 11887317 TI - Heavy metals in water bodies purified by suspended substrate of rivers. AB - The equations, which are used to describe the relationships of adsorption quantity (s), adsorption percent (Pa), aqueous equilibrium concentration(c) of heavy metal on river suspended substrates and the ratio of adsorbent to water(j), are developed when heavy metal adsorption on river suspended substrate satisfies with linear adsorption equation. The results, according to the simulation from heavy metal adsorption on suspended substrates of several main Chinese rivers from a previous research report, indicated that these developed equations could describe the linear adsorption processes in practice very well, meanwhile, the adsorption equilibrium constant of adsorbent for heavy metal was an intensity factor regardless of ratio of suspended substrates to water but strongly depended on media's pH. Furthermore, the suspended substrates of Yellow River gave stronger purification ability for Pb than for Cd and Cu. When Cd was purified by different river suspended substrates, it exhibited that the order of their purification ability for Cd was that of Songhuajiang River > Zhujiang Pearl River > Yellow River, which was consistent with their contents of cation exchange capacity(CEC). In addition, we estimated and compared the purification ability of river suspended substrates for cadmium, and the resulting purification percent was 37.64%, 64.58% and 50.98% for Songhuajiang River, Yangtze River and Zhujiang River, respectively. PMID- 11887318 TI - Double catholyte electrochemical approach for preparing ferrate-aluminum: a compound oxidant-coagulant for water purification. AB - Ferrate is an excellent water treatment agent for its multi-functions in oxidation, disinfection, coagulation and adsorption, but its coagulation ability depends on its dosage and is after its oxidation. This paper focuses on preparing a new kind of ferrate combined with alum to enhance its coagulation function for water purification. An effective electrolysis reactor was designed and employed in the test. Some key parameters in the process of electrolysis concerning the preparation efficiency, such as the current density, temperature and alkalinity were also investigated. The proper conditions for ferrate-alum preparation were determined. Under the condition of 5 V given voltage, 6 h electrolyzing interval, below 2% alum concentration (in weight), a combined liquid ferrate-alum products was successfully prepared, which contained 0.0294 mol/L FeO4(2-) and 0.0302 mol/L total soluble ferron with 2% Al2O3. There was no insoluble ferron produced by controlling an optimum electrochemical condition. PMID- 11887319 TI - Cyanobacterial flora and the physico-chemical environment of six tropical fresh water lakes of Udaipur, India. AB - The cyanobacteria and physico-chemical environments of six tropical fresh water lakes of Udaipur, India were investigated. These lakes receive varying nutrient inputs from different sources. Altogether 51 species of cyanobacteria were recorded. Species composition varied between lakes and between seasons. Lake VI (Baghdara), which receives nutrients from natural sources only, differed considerably from the others in water chemistry and composition of dominant species. Lake II (Swaroop Sagar), eutrophied due to sewage inputs, was species poor. Non-diazotrophs, represented by 27 species, dominated during summer. With few exceptions, N2-fixing species, both heterocystous and unicellular diazotrophs (represented by 24 species), were dominant during winter. Microcystis aeruginosa, Phormidium sp. and Anabaena flos-aque were the dominant taxa of lakes characterized by sewage eutrophication. The study shows that both species diversity and community composition were affected by water chemistry. PMID- 11887320 TI - Concurrency: a system design approach to environmental management and sustainability. AB - The lessons of history indicate that mismanagement of natural resources and the environment often leads to potentially adverse consequences. The increasing interest in economic development, particularly in the developing countries of the world coupled with increasing population pressures and the globalization of economic activity is placing noticeable stresses on the ultimate sustainability of both human and environmental systems. Sustainable development is not a new concept. It has been an area of concern for different elements of society for some time. Yet efforts to understand the implications of sustainable development have not, until recently, been formalized. We have focused singularly on economic development and environmental quality as if they were mutually exclusive. This paper focuses on the concept of concurrency as both a conceptual framework and practicable method of understanding and implementing the ecology and economy of sustainability. PMID- 11887321 TI - Chemical composition of aerosols in winter/spring in Beijing. AB - In 1999 aerosol samples were collected by cascade at Meteorological Tower in Beijing. The 12 group aerosol samples obtained were analyzed using PIXE method, which resulted in 20 elemental concentrations and size distribution of elemental concentrations. From the observation, the elemental concentrations, size distribution of elemental concentrations and their variations are analyzed. It shows that concentrations of the most elements in aerosols increase greatly compared with those in the past except that the concentrations of V, K, Sr, and the source of aerosols has changed greatly in the past decade. Fine mode aerosols increase more rapidly in the past decade, which may be due to the contribution of coal combustion and automobile exhaust. Pb content in aerosol is much higher than that at the beginning of 1980s, and has a decreasing trend in recent years because of using non-leaded gasoline. PMID- 11887322 TI - Calculation method of quantum efficiency to TiO2 nanocrystal photocatalysis reaction. AB - The quantum yield is an important factor to evaluate the efficiency of photoreactor. This article gives an overall calculation method of the quantum efficiency(phi) and the apparent quantum efficiency(phi a) to the TiO2/UV photocatalysis system. Furthermore, for the immobility system (IS), the formulation of the faction of light absorbed by the TiO2 thin film is proposed so as to calculate the quantum efficiency by using the measured value and theoretic calculated value of transmissivity (T). For the suspension system(SS), due to the difficulty to obtain the absorption coefficient (alpha) of TiO2 particulates, the quantum efficiency is calculated by means of the relative photonic efficiency (zeta r) and the standard quantum yield (phi standard). PMID- 11887323 TI - Assessment of water pollution control strategies: a case study for the Dianchi Lake. AB - Lake eutrophication has increasingly become a major environmental issue in China. Although significant efforts have been made towards its resolution in the last decade, most of the implemented control strategies are fragmented, and the formation of policy lacks of sound scientific basis and long-term objectives. Taking the well-known Dianchi Lake as a case study, this paper presented a comprehensive assessment for the effectiveness of various eutrophication control strategies. It is expected that the concluding lessons would have a major implication to future eutrophication control. PMID- 11887324 TI - Trace metals distribution in Synodontis membranaceus, sediments, Asystasia gangetica and Platostoma africanium from Ofuafor River around Delta Glass Factory in Ughelli North Local Government Area, Delta State Nigeria. AB - The trace metals analysis in synodontis membranaceus (head and tail), bottom sediments, Asystasia Gangetica and Platostoma Africanium were carried out using atomic absorption spectrometer of model Perkin Elmer 3110. Metals analysed were copper, nickel, mangenese, chromium, iron lead and cobalt. These metals were detected in the above samples. Vegetation samples concentration in copper, manganese, chromium, iron and cobalt were higher than those obtained in bottom sediments. The tail part of the fish contents of trace metals were also higher than those of the head. The results obtained in this work exceeded the results of water analysis carried out by Omoregha on the same river. Metals such as copper, nickel, manganese and chromium were below detection limit in the water from the same river. The bioaccumulation of these trace metals in these samples were traced to activities of Delta Glass Factory. PMID- 11887325 TI - Sustainable development pattern and the strategy in the three Gorges Reservoir Areas. AB - The development pattern, development situation, and existing problems of land exploitation in Zigui County, Three Gorges Reservoir Areas of China were presented. The sustainable development mode and its strategy in the Three Gorges Reservoir Areas was also discussed. A sustainable development framework for low mountain regions, middle mountain regions and high mountain regions was developed, and management countermeasures for structural optimization of complex ecosystems were advanced. PMID- 11887326 TI - Dynamic analysis and assessment for sustainable development. AB - The assessment of sustainable development is crucial for constituting sustainable development strategies. Assessment methods that exist so far usually only use an indicator system for making sustainable judgement. These indicators rarely reflect dynamic characteristics. However, sustainable development is influenced by changes in the social-economic system and in the eco-environmental system at different times. Besides the spatial character, sustainable development has a temporal character that can not be neglected; therefore the research system should also be dynamic. This paper focuses on this dynamic trait, so that the assessment results obtained provide more information for judgements in decision making processes. Firstly the dynamic characteristics of sustainable development are analyzed, which point to a track of sustainable development that is an upward undulating curve. According to the dynamic character and the development rules of a social, economic and ecological system, a flexible assessment approach that is based on tendency analysis, restrictive conditions and a feedback system is then proposed for sustainable development. PMID- 11887327 TI - Intersexual plasticity in aspects of the biology of the mudskipper Periophthalmus barbarus (Gobhdae) in the mangrove swamps of IMO Estuary, Nigeria. AB - Between April 1992 and March 1993, intersexual plasticity in aspects of the biology of the mudskipper, Periophthalmus barbarus was studied. Sex was differentiated on the basis of the genital papillae, it was broader in females than males. Sexually active females were slightly heavier than similarly-sized males. Sex ratio was strongly female-biased. Length-weight relationship of the sexes was isometric. There was no significant difference in the feeding intensity of both sexes; uniformity was also apparent in diet breadth. There was no named sex-based difference in hepatosometic index but condition index was higher in males than females. The uniformity in intersexual plasticity of P. barbarus is consequent upon homogeneity of the mangrove ecosystem, which can only be altered anthropogenically. PMID- 11887328 TI - Changes in immunological characteristics of white blood cells after administration of standardized mistletoe extract. AB - After administering standardized mistletoe extract, Viscum album L, (Iscador injections of 0.1 mg twice and 1.0 mg in defined intervals) the functional characteristics of microcirculation and immunological behavior of the white blood cells in different target tissues (derma, intestine) were investigated in healthy volunteers by vital microscopic investigation over 13 days of observation. The investigations showed a temporarily improved function of the microcirculation and an increased adhesion and transmigration of white blood cells in the target tissue areas. This observation was evaluated as a biologically relevant immunomodulation. Further investigations under pathophysiological conditions with regard to complementary administration of the test substance (e.g. to cancer patients) appear promising. PMID- 11887329 TI - Raloxifene: another selective estrogen modulator. AB - The role of estrogens as one of the prime stimulators of tumor cell proliferation is well recognized; efforts to interfere with the initiation and promotion of breast and other cancers by endocrine manipulation have a long and successful past. The benzothiophene derivate Raloxifene is a relatively recent newcomer into a heterogeneous family of compounds loosely called antiestrogens, which implies their ability to act as antagonists to estrogen effects via competitive binding to various steroid receptors. This is a reductionist explanation, since their action is colorful and varied; they interact with lipid transduction cascades, covalently bind to DNA and to different proteins and regulate growth factors and the expression of various genes, such as erB2, mdr1 and p53; they complex with E cadherin/catenin and are thus able to induce apoptosis, actively or indirectly. Raloxifene not only modulates estrogen effects, but has been found to reduce bone demineralization and atherogenesis, without carcinogenic stimulation of the endometrium. PMID- 11887330 TI - Dynamics of chronic active herpesvirus-6 infection in patients with chronic fatigue syndrome: data acquisition for computer modeling. AB - Ten adult patients with persistent active HHV-6 variant A infection and clinical chronic fatigue syndrome (CFS) were studied over a period of 24 months after initial clinical diagnosis. CFS was diagnosed according to IIIP-revised CDC criteria as defined by the CFS Expert Advisory Group to the German Federal Ministry of Health in 1994. Changes in HHV-6 antibody titer, viral DNA load, peripheral blood T lymphocytes and subpopulations, as well as CD4/CD8 cell ratio and cell death (apoptosis) were monitored. Data were collected for comparison with respective changes in acute HHV-6 infection and as a basis for future computer simulation studies. The results showed variable but slightly elevated numbers of HHV-6 DNA copies in the blood of patients with CFS, while PBL (peripheral blood lymphocyte) apoptosis rates were clearly increased. CD4/CD8 cell ratios varied from below 1 up to values as seen in autoimmune disorders. Contrary to acute HHV-6 infection, T lymphocytes do not exhibit the usual response to HHV-6, that is elevation of mature and immature populations suggesting a certain degree of unresponsiveness. The data suggest that persistent low-dose stimulation by HHV-6 may favor imbalanced immune response rather than overt immune deficiency. This hypothesis requires confirmation through additional functional studies. PMID- 11887331 TI - Species susceptibilities to chemical carcinogenes: a critical appraisal of the roles of genetic and viral agents. AB - The carcinogenesis literature concerning the susceptibilities of the various animal species and humans to chemical carcinogens is reviewed. The fundamental issue is whether the different laboratory animal species used in the past and at the present are the appropriate systems to detect the carcinogenic properties of chemicals. It was concluded that the main problem is the presence of high to moderate incidences of spontaneously-appearing cancers in laboratory animals. These tumor incidences are, to a significant extent, either caused by genetic factors and/or viruses which render the systems excessively-susceptible to the carcinogenic action of a large proportion of chemicals. The same types of genetic factors and/or viruses do not exist in the human species. Consequently, the data obtained in the various animal species are exaggerated to an unrealistic proportion due to the inherent flawed designs, therefore, it has little bearing in the human species. It is further concluded that a new animal breeding program should be initiated with the specific aim of developing several tumor-free or near tumor-free species and strains of laboratory animals. Until then, a healthy state of mind is required to separate the reality from the fiction. PMID- 11887332 TI - Comparative dermal pharmacology and toxicology of 4,4'-dihydroxybenzophenone-2,4 dinitrophenylhydrazone (A-007) in rodents and primates. AB - 4,4'-Dihydroxybenzophenone-2,4-ditrophenylhydrazone (A-007) has demonstrated anticancer activities, when administered topically to patients with metastatic cancer to the skin. Acute, subacute and subchronic dermal studies with A-007 in adult rabbits, rats, guinea pigs and monkeys failed to demonstrate local or systemic toxicity when applied topically as a 0.25% gel. A-007 did not penetrate the dermal lymphatics and did not produce detectable levels of A-007 in the plasma when applied as a 0.25% gel topically to skin. In the above studies, topically administered A-007 stimulated local sub-epithelial and dermal lymphocyte modulation, with increased CD8+ cytotoxic lymphocytes (CTL) noted, in guinea pig skin. Generally topical A-007 is well tolerated and may have useful immune modulation properties. PMID- 11887334 TI - In vivo modulation of ETS genes induced by electromagnetic fields. AB - We have previously shown that electromagnetic field (EMF) exposure induces ETS1 oncogene overexpression in different cell lines. In order to investigate in vivo EMF effects, BALB/c mice were exposed at different times to 50 MHz radiation, modulated (80%) at 16 Hz. The exposed and control animals were sacrificed and the spleen excised for rt-pcr and western blot analysis. We observed an increase in ETS1 mRNA and protein expression, but a decrease in ETS2 protein levels. Preliminary results from this experimental model show in vivo evidence of the effect of EMF on ETS oncogene expression. PMID- 11887333 TI - Suitable indices for evaluating the intensity of tumor metastasis in a mouse experimental metastatic model. AB - We investigated the time course of developmental changes of tumor metastasis in mice intravenously inoculated with B16-F10 melanoma cells. The mice showed a normal blood profile at 7 and 14 days after the injection, while at 21 days they developed severe anemia and thrombocytopenia. The number of metastatic lung nodules in the mice at 7 and 14 days after the cell injection was 68 +/- 12 and 326 +/- 39 (mean +/- S. E. of 5 mice), respectively, while it could not be accurately counted at 21 days owing to a fusion of the nodules. The wet weight and the melanin content of the lung markedly increased at 21 days. These results suggest that an abnormal blood profile, lung weight and the melanin content of the lung are suitable indices for the late stage of the experimental metastatic model, while the number of lung nodules is suitable for the early stage. PMID- 11887335 TI - Effects of chronic treatment with caffeine on behaviour and related parameters in male and female mice. AB - The effects of chronic free access to caffeine (0.01% or 0.05%) in drinking water and subsequent withdrawal on spontaneous motor activity for 24 hours and some related parameters were examined in 8-week-old male and female ICR mice. In the males, the 0.01% group showed little response, but in the 0.05% group the activities in both light- and dark-phases and, consequently, in total increased and peaked on day 5 of treatment. The response gradually decreased on days 15 and 30 and reached the control level after 30 days of caffeine withdrawal. Meanwhile, in the females, the activity was stimulated by both 0.01% and 0.05% of caffeine, at the dark- and light-phases in the former and latter, respectively. The response peaked at 30 days and decreased near to the control level thereafter in both groups. Caffeine affected little the food intake; however, water intakes were higher and lower than the control in the 0.05% and 0.01% male groups, respectively, but the opposite was true in the females. Plasma component levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), cholesterol and glucose were higher than the control in the males and females treated with 0.05% of caffeine. The caffeine had little effect on the body weight change, organ weights and external appearance throughout the experiment. Thus, the sex- and dose-related differences in the responses to caffeine of spontaneous motor activity and related parameters were proved under physiological conditions. PMID- 11887336 TI - Effects of gonadectomy at different ages and Sasa Health, bamboo grass leaf extract, on spontaneous motor activity in female and male mice. AB - To investigate the role of gonadal hormones in spontaneous motor activity and the contribution of Sasa Health, bamboo grass leaf extract, to this process, female and male ICR mice were gonadectomized at 2 (Experiment I) or 9-12 months of age (Experiment II). The spontaneous motor activity was measured 1 week and 1 month after the operation, then after a subsequent 1 month of treatment with Sasa Health administered in the drinking water. Ovariectomy caused a significant decrease in activity even after 1 week in Experiment I, but had little effect in Experiment II. In the males, a significant decrease was found only in the dark phase 1 month after castration in Experiment I, while little change in activity was induced by castration in Experiment II. Administration of Sasa Health caused an increase and decrease in the activity of the females and males, respectively, of both the control and the gonadectomized groups in Experiment I. Sasa Health had little effect on the motor activity in Experiment II. The agent acted on the plasma levels of several components modulated by gonadectomy, mostly restoring them to the control values. These findings indicate that gonadal hormones and Sasa Health influence spontaneous motor activity and plasma component levels in both sexes and have apparent sex- and age-related effects. PMID- 11887337 TI - Dynamics of active progressive infection with HIV1: data acquisition for computer modeling. AB - Nineteen adult patients with progressive HIV1 infection, which progressed within 5 years from acute HIV syndrome to final AIDS were studied. Changes in HIV antibody titer, viral RNA load, peripheral T lymphocytes and subpopulations as well as CD4/CD8 cell ratio and cell death (apoptosis) were monitored. The data were collected for comparison with HHV-6 infection, which involves the same cell populations yet patients usually recover, and to serve as a further basis for future computer simulation studies. The results showed progressive increases of viral RNA copies in the patients' plasma even during clinical latency, which correlates with lymphocyte apoptosis and CD4 cell loss. Besides apparent direct CD4 cell destruction, there was indication of a disturbed intrathymic T cell differentiation. Pathologic cell changes in HIV infection continue until final death of the patient and do not return to normal after variable times as in HHV-6 infection. While HHV-6 infection can serve as models for immunostimulation, with or without immune dysregulation in computer simulation studies, HIV infection is a model for immunostimulation with final immune deficiency and cellular aplasia. PMID- 11887339 TI - Cervical intraepithelial dysplasia (CIN 3) and history of herpes genitalis (HSV2) in women living in Attica, Greece. AB - This article presents a study on women living in Attica, Greece (an area of 4.5 million inhabitants) who visited the outpatient clinics of two specialized hospitals for a follow-up for cervical intraepithelial neoplasia (CIN 3). A random sample of 170 women with CIN 3 and a control group of 195 women coming from the general population were interviewed. A semi-structured designed questionnaire was used as an instrument for data collection. The aim of the study was to search for a possible correlation between CIN 3 and a past history of herpes genitalis (HSV2) of the women and/or their partners. According to the results, CIN 3 is more frequent among women between 26 and 40 years of age. The majority of the patients (70.3%) reported a past history of HSV2, but only 10.9% of the subjects in the control group had the same medical history (p = 0.000). Sixteen percent of the patients and only 5.3% of the subjects in the control group reported infection of their sexual partners with HSV2. 75 patients (44.6%) stated that they did not know if their sexual partners had a past history of HSV2, while only 27 women (13.3%) in the control group reported the same (p = 0.000). Patients reported more sexual partners than women in the control group. PMID- 11887338 TI - Antimicrobial activity of prenylflavanones. AB - Among two flavanones [YS01, YS02] and eight prenylflavanones [YS03-YS10], preliminary screening with fifteen test bacterial strains showed that YS06 was the most active agent. YS06 exhibited highly significant antimicrobial action when tested against 228 bacterial strains comprising two Gram-positive and six Gram-negative genera. The in vitro susceptibility test was carried out by determining the minimum inhibitory concentration (MIC) of YS06 by agar dilution technique. Twenty-two out of fifty strains of Staphylococcus aureus were inhibited at 25 to 50 micrograms/mL of the agent. YS06 also inhibited strains of Salmonella, Shigella and a few strains of Escherichia coli strains were also highly sensitive to YS06, while Kleibsiella spp. and Pseudomonas aeruginosa were much less sensitive. In in vivo study, YS06 offered significant protection (p < 0.001 according to chi-square test) to Swiss albino mice (challenged with 50 minimum lethal dose (MLD, virulent bacterium) at concentrations of 160 and 80 micrograms/mouse. PMID- 11887340 TI - Soluble low-molecular-mass heat shock proteins and tumor-associated antigens in prevention and therapy of chemically-induced cancers. AB - There is increasing evidence that tumors express putative target molecules in a therapeutic immune reaction. Identification of immunogenic tumor-associated antigens (TAA) may enable the development of new modes of vaccination with the addition of immunotherapy as a potentially powerful weapon in the fight against cancer. In the present review, the authors' observations on the role of the soluble low-molecular-mass heat shock proteins and tumor-associated antigens, named as a complex of STAA, in the prevention and therapy of chemically-induced tumorigenesis are analyzed and compared with data from the literature. It has been shown that STAA have both tumor-preventive and tumor-suppressive effects on chemically-induced cancers of the colon, skin and mammary glands in rats and mice. These effects were shown to be connected with activation of the host's immune system, especially that which is responsible for the activity of T and B lymphocytes. These findings have led to a wave of new trials involving cancer immunotherapy. Understanding the mechanisms of antitumor immunity and identifying relevant tumor-specific antigens is expected to improve vaccine strategies and provide for a successful cancer therapy in the future. PMID- 11887341 TI - Paternal smoking during pregnancy and the risk of childhood brain tumors: results of a meta-analysis. AB - OBJECTIVE: Prior epidemiological studies suggest a possible association between paternal smoking during pregnancy and risk of childhood brain tumors (CBT). A meta-analysis was performed statistically pooling all available observational studies on this topic in order to evaluate this suspected association. METHODS: Using previously described methods, a protocol was developed for a meta-analysis examining the association between paternal smoking during pregnancy and subsequent development of primary brain tumors in their offspring. Literature search techniques study inclusion criteria and statistical procedures were prospectively defined. Data from epidemiological studies were pooled using a general variance based meta-analytic method employing confidence intervals previously described by Greenland. The outcome of interest was a summary relative risk (RRs) reflecting the risk of childhood brain tumor development associated with father's smoking during the index pregnancy. Sensitivity analyses were performed when necessary to explain any observed statistical heterogeneity and/or to evaluate the impact of demographic or study characteristics on the summary estimate of effect. RESULTS: Seven observational studies meeting protocol specified inclusion criteria were obtained via a comprehensive literature search. These studies enrolled a total of 3,600 patients. Analysis for homogeneity demonstrated that the data were homogeneous (P = 0.52) and could be statistically combined. Pooling all seven reports yielded a RRs of 1.29 (1.07-1.53), a statistically significant result suggesting a 29% increased risk of brain tumor development associated with paternal smoking during pregnancy. An analysis of father's smoking impact on CBT risk based on "ever" versus "never" smoking history gave a RRs of 1.14 (0.98-1.34), a marginally non-statistically significant result. CONCLUSION: The available epidemiological data suggest an association between paternal smoking during pregnancy and pediatric brain tumor development. Although this association is biologically plausible, limitations in study designs limit definitive conclusions based on available data. PMID- 11887342 TI - [Lithium thyrotoxicosis. Report of a case and review of the literature]. AB - It is known that lithium therapy causes goiter and hypothyroidism in about 8% of cases, particularly in patients with circulating antithyroid antibodies. Rarely, lithium is associated with hyperthyroidism: the relationship seems not to be casual, since hyperthyroidism in these patients is three times that of the normal population. These subjects are affected by a diffuse toxic goiter, with or without ofthalmopathy (the majority), or by multinodular toxic goiter or by a painless thyroiditis. In the first case, lithium acts as an immunostimulating factor, even if the real prevalence of antithyroid stimulating antibodies is not well known in these patients; in the second case, an "escape" mechanism following the hormonal release inhibition, favoured by lithium, can explain hyperthyroidism. In the third case, as in this here described, in which no thyroid map is seen at the scintiscan, a local, inflammatory mechanism is involved, in a similar way of some amiodarone induced thyrotoxicosis. PMID- 11887343 TI - [External otitis]. AB - Otitis externa is one of the most common diseases in ORL practice, during summer; the treatment of otitis externa may be simple and easy or protracted and frustrating, also with fatal outcome. Many local factors may interfere with the normal defences against infections in the external auditory canal. Removing or dissolving the cerumen by water or other instruments eliminates an important barrier to infections: its acids inhibit the growth of bacteria (Staphylococcus aureus and Pseudomonas aeruginosa) and fungi (Aspergillus). Also skin abrasions or irritation, allergic diseases and many systemic condition like anaemia, vitamin deficiency, endocrine disorders (diabetes) and various forms of dermatitis cause a lower resistance to infections in external auditory canal. Even if the prognosis remains benign in the majority of cases, important complications could appear like: malignant otitis externa, facial nerve paralysis, tympanic bone osteomyelitis, pericondrytis. Successful treatment depends on a proper diagnosis and therapy: the most important factor in the treatment is repeated debridement of the external auditory canal by the physician. The use of Castellani' Tintura rubra, hydroalcoholic solution of phenic fuchsin, can be very effective for bacteria and mycotic eradication. Culturing of ear canal infection could be performed on the second or third visit if the otitis externa is not responding to therapy. Complication are not frequent, but malignant otitis externa can be mortal. Dermatological consultation is often necessary for correct diagnosis. PMID- 11887344 TI - [Dynamics of communication between physician and the oncologic patient]. AB - The informed consent of patients is ethically and legally required for all medical practice. The disclosure of information to cancer patients is one of the most important issues mostly in oncological practice. Because most patients now want to know the truth about their diagnosis and prognosis, the ability to discuss the cancer diagnosis, disease recurrence, or treatment failure, and to solicit patients' views about resuscitation or hospice care, are important verbal skills for oncologists and other oncology care providers. If clinicians involved in cancer care are able to well communicative with patients, these ones and their families can achieve a good level of quality of life and psychological adjustment. Moreover, the ability to clearly articulate a treatment plan or elicit patient preferences for treatment are prerequisite to informed consent. Despite these imperatives, clinicians do not routinely receive training in key communication skills that could enable them to accomplish these tasks. Disclosure of information needs to be individualized, and social and cultural background has to be taken into consideration. This review considers deficiencies in the conduct of key communication tasks and their consequences. The reasons for these deficiencies are explored and guidelines to resolve these issues and to further new alternative methods are proposed. PMID- 11887345 TI - [New and old beta-blockers in the treatment of heart failure]. AB - Controlled clinical trials, performed in more than 16,000 patients to date, have consistently shown the beneficial effects of long-term beta-blocker therapy in patients with chronic heart failure. However, it is not clear whether this represents a class effect or it is specific only to some agents. Beneficial effects on the prognosis of the patients with mild to moderate heart failure have been shown with metoprolol, bisoprolol, and carvedilol. However, these beta blockers differ in their pharmacological characteristics. Metoprolol and bisoprolol are selective for beta 1-adrenergic receptors and are devoid of ancillary properties. Carvediol, at doses of 50 mg daily, blocks all beta 1-, beta 2-, and alpha 1- adrenergic receptors, and has associated antiproliferative and antioxidant activities. These differences cause a different acute hemodynamic response with a reduction in cardiac output and a tendency to a rise in pulmonary wedge pressure with selective agents and no change in cardiac output and a slight decrease in pulmonary pressures with carvedilol. Accordingly, when the therapy is started, the most frequent side effects are worsening heart failure with metoprolol and bisoprolol and hypotension and dizziness with carvedilol. It is still controversial whether these differences may also influence the long-term effects of therapy. Differently from selective beta-blockers, carvedilol does not upregulate beta 1-receptors, blocks all adrenergic receptors, decreases cardiac norepinephrine release, thus providing a more comprehensive blockade of the cardiac adrenergic drive. These properties have caused a larger increase in LV function and a lack of improvement in maximal exercise capacity with carvedilol, compared to selective beta-blockers. It is however, unclear whether these differences may also influence the patients' outcome. PMID- 11887346 TI - [Glutathione and N-acetylcysteine in the prevention of free-radical damage in the initial phase of septic shock]. AB - The hyperproduction of oxygen free radicals (OFR) and the weakening of natural scavenging mechanisms are implicated in endothelial damage and in multiple organ failure during septic shock. Many authors have proposed the use of antioxidants to decrease OFR damage. Glutathione (GSH) is one of the most important endogenous antioxidants. It plays the role of a sulphydryl group provider for scavenging reactions. N-Acetylcysteine (NAC) is an artificial precursor of GSH which both increases GSH levels and acts as OFR scavenger. The authors carried out a clinical trial to confirm the capacity of high doses of GSH and NAC to cooperate in reducing lipoperoxidative damage in patients with early septic shock. Patients were divided into three groups who received shock therapy only or shock therapy plus GSH or shock therapy plus GSH plus NAC. OFR damage was evaluated by measuring expired ethane, plasma malondialdehyde, complement activation and clinical scores. The study demonstrated that the group who received GSH and NAC showed a significant decrease in peroxidative indexes and an improvement of the clinical scores if compared with the other two groups. In conclusion the administration of high doses of NAC added to GSH significantly decreases the peroxidative stress of patients with early septic shock. PMID- 11887347 TI - [Bronchial endoscopy in respiratory disease]. AB - Less than 50 years after the birth of the fiberoptic bronchoscope, bronchial endoscopy had had a considerable progress and has found a wide diffusion of its applications to many respiratory diseases. The author briefly explains the most important and used applications, wishing this diagnostic therapeutical method will soon become an independent discipline. PMID- 11887349 TI - [Prospects of molecular therapy in breast carcinoma]. AB - Recent studies on genetics and molecular biology have allowed the discovery of new therapies for breast cancer. Some possibilities of therapy of this tumor emerged from these discoveries are briefly discussed. PMID- 11887348 TI - [Role of hemostatic markers in the pathogenesis of coronary diseases]. AB - The underlying processes that lead to atheroma formation and coronary thrombosis are complex and still incompletely understood. There is a growing interest in defining a possible role of haemostatic markers for atherothrombotic risk. There is a large body of literature concerned with common genetic variations within genes of haemostatic factors, their plasma levels, and how they might influence the development of arterial thrombotic diseases. The aim of this review is to assess our current state of knowledge on this topic. PMID- 11887350 TI - Cost-effectiveness of enoxaparin as thromboprophylaxis in acutelly ill medical patients from the Italian NHS perspective. AB - OBJECTIVE: To determine the cost-effectiveness of thromboprophylaxis with enoxaparin versus no thromboprophlaxis in patients with acute medical illness in Italy from the NHS perspective. METHODS: Markov process analysis techniques were used to model the health economic outcomes. Clinical data were derived mainly from the MEDENOX trial, while health care utilization was derived from Delphi panels. RESULTS: An analysis over the MEDENOX trial period shows that the cost per event avoided is [symbol: see text] 2450.99, while the cost per life saved is [symbol: see text] 8395.89. The lifetime model, which assumes no higher risk for recurrence of VTE and mortality in asymptomatic patients, shows that the use of enoxaparin leads a cost per event avoided of [symbol: see text] 2243.20, and cost per life year gained of [symbol: see text] 605.38. The lifetime model, which assumes a higher risk for recurrence of VTE in asymptomatic patients, shows that enoxaparin is dominant over no thromboprophylaxis. CONCLUSION: The results showed that the favourable clinical benefit of enoxaparin as thromboprophylaxis in patients with acute medical illness, which was observed in the MEDENOX trial, results in a positive health economic benefit on the short-term and long-term in the health care setting of Italy. PMID- 11887351 TI - [Polymicrobial infective endocarditis in Italy]. AB - In our series, including 1053 cases of infective endocarditis (IE) collected from 86 Infectious Diseases Centres in Italy between 1984 and 1999, we identified 34 cases (3.2%) with a polymicrobial etiology. Intravenous drug abuse was the most important risk factor for the development of polymicrobial IE. Twenty three patients had a left sided-IE and 6 patients had a right-sided IE. The most commonly encountered microorganisms were Staphylococci and Streptococci and the most frequently observed associations of microorganisms were those between Staphylococci and Gram-negative bacteria and between Staphylococci and fungi. Twelve patients (35.3%) underwent surgery, and 5 patients (16.7%) died. Polymicrobial endocarditis did not differ clinically from IE caused by a single microorganism, and the prognosis seems to be related to the site of infection and to some specific pathogens. PMID- 11887352 TI - [Expression of the costimulatory CD28 antigen on peripheral blood CD8+ T lymphocytes in patients with chronic hepatitis C]. AB - Virus-specific activation of the CD8+ cytotoxic T lymphocytes, which may affect the extension of histopathological damage in chronic hepatitis C, is closely related to the role of the so called "costimulatory antigens", particularly CD28, allocated on the surface of the T lymphocytes. This study evaluated the display of CD28 antigen on circulating T lymphocytes CD8+ in patients with chronic hepatitis C. The eventual correlation between CD28 expression, viral load, aminotransferase levels and degree of histological damage was also studied. According to our data, the prevalence of circulating TCD8+ CD28+ cells didn't differ among anti-HCV positive, HBV positive subjects and healthy controls. In HCV-positive subjects, prevalence of TCD8+ CD28+ cells was higher in patients with chronic persistent hepatitis than in chronic active cases with a significant negative correlation with the Knodell histological "score". Further studies are required to ascertain the role of CD28+ expression in those subjects with severe hepatic histological damage due to chronic hepatitis C. PMID- 11887354 TI - Advocating for the oral health of children. PMID- 11887353 TI - Anaphylaxis update. PMID- 11887355 TI - Helping those who hurt themselves. PMID- 11887356 TI - The school nurse perspective on self-injury. PMID- 11887358 TI - The obesity epidemic--what are we to do? PMID- 11887357 TI - Character building. PMID- 11887360 TI - Invisible pervasive handicap in the classroom. PMID- 11887359 TI - Helping children and teens cope with traumatic events and death: the role of school health professionals. PMID- 11887361 TI - Mental health approaches to violence prevention in schools. PMID- 11887362 TI - Peer review. PMID- 11887363 TI - A-dec as a partner to dentists. AB - A-dec has become the largest manufacturer of dental equipment in America by focusing on the customer, the employee, and the product rather than the bottom line. The driving principles of the company are (a) quality comes first, (b) customers are the focus of everything, (c) continuous improvement, (d) employee involvement is a way of life, (e) dealers and suppliers are our partners, and (f) integrity is never compromised. PMID- 11887364 TI - Oral care clinical trials at Hill Top Research. AB - Oral health care products generally require laboratory or clinical testing prior to being introduced to the market. Companies that develop such products have three options for such testing--their own facilities, dental schools, or clinical research organizations (CROs). Laboratory or clinical trials involving subjects can be conducted to test claims of safety and efficacy as well as cosmetic and therapeutic claims. CROs, which conduct such research on a fee-for-service basis in an independent environment, are an attractive alternative in many cases. PMID- 11887365 TI - Locum tenens--a concept it is time for dentistry to embrace. AB - As shown by success in other health professions, dentists can effectively stand in for other dentists who need to be absent from their offices for reasons of health, family matters, education, and many similar reasons. Locum Tenens agencies can facilitate such temporary substitutions and are especially valuable in maintaining productive use of fixed overhead. Examples of situations where a Locum Tenens arrangement may be beneficial include practice transitions necessitated by health problems, educational opportunities, temporary leaves due to pregnancy or other family matters, wellness interventions, and management of dentist shortage areas. PMID- 11887366 TI - Helping dentists manage accounts receivable. AB - First Pacific Corporation (FPC) has worked with dental practices since 1961, providing personal services that optimize practice performance. In addition to being the premier service provider for administrative tasks in dental offices, they supply state-of-the-art hardware and accounts receivable management software. FPC designs and teaches practice development strategies, deliver on site training, and much more. FPC is dedicated to the long-term professional success of dental clients, their staff, and their practices through a unique, integrated package of services. As a family-owned business, with headquarters in Salem, Oregon, FPC employs approximately two hundred staff who serve practices in twenty-two states. PMID- 11887367 TI - Making panographic equipment available in the office. AB - Panoramic Corporation began as a business leasing panographic radiographic equipment to dentists in the Fort Wayne, Indiana, area and has become the largest manufacturer of such equipment in the world. While ensuring a quality product, Panoramic has built its success on making this high-end technology available to dentists with minimal risk and a high level of maintenance. Direct marketing and lease programs have been keys to making panographic capabilities available to many dentists. PMID- 11887368 TI - Les Schwab tire centers leads the way in dental insurance. AB - A family-owned retail tire company has a successful history with direct reimbursement for dental and other health care benefits. Although there is a co pay and some elective services are not covered, employees are not asked to fund insurance premiums and freedom of choice with regard to provider is ensured. This approach is grounded in the family orientation of the company. PMID- 11887369 TI - Universal patient acceptance: ethics pipe dream or key to improved access in dentistry? AB - The authors discuss ways that dentistry engages silently and sometimes unknowingly in practice patterns that adversely affect public access to dental care. The concept of acceptance is explained and contrasted with treatment and with access to care. The concept of Universal Patient Acceptance (UPA) is introduced, with a focus on how it underlies and precedes access, creating a pathway so that truer universal access to dental care can be realized. The authors argue that a commitment to Universal Patient Acceptance shows promise as an important starting point in the dental profession's concern to address society's unmet oral health needs. PMID- 11887370 TI - Red beads and funnels. AB - Dentists, like most managers, believe that unanticipated team results are evidence of poor performance on the part of employees. While this can be the case, it is much more likely that most variation is inherent in the system and is probably not under the control of staff. The dentist, as the manager, has full control and full responsibility for guaranteeing that the office runs effectively and for improving its operation. PMID- 11887371 TI - The American College of Dentists: a vision in action. PMID- 11887372 TI - Oral health and social justice: leadership opportunities for dentistry. PMID- 11887373 TI - The influence of different facial components on facial aesthetics. AB - Facial aesthetics have an important influence on social behaviour and perception in our society. The purpose of the present study was to evaluate the effect of facial symmetry and inter-ocular distance on the assessment of facial aesthetics, factors that are often suggested as major contributors to facial aesthetics. The material consisted of 36 standardized facial photographs of patients (10-17 years of age) attending the orthodontic department. All except one were Caucasian. Eleven of these photographs (all Caucasian) were either left in their original form or modified by: (1) enlargement of the inter-ocular distance by 20 per cent; or (2) mirroring the right part of the face over the left part to obtain symmetry. Three series of 36 slides were composed, each including 11 modified slides or their unmodified counterparts. These were evenly distributed over the three series using a Latin square design. The photographs were assessed for their facial aesthetics using a visual ratio scale by a panel of 50 undergraduate dental and law students. The 11 modified/unmodified photographs enabled calculation of the effect of the modifications undertaken on the assessment of facial aesthetics. The scores of the 25 remaining photographs in each series were used as baseline data in order to estimate the inter- and intra-observer reproducibility. MANOVA and post-hoc t-test revealed significant differences between the unmodified and modified photographs (P < 0.001). The results show that symmetry and inter-ocular distance enlargement have a negative effect on facial aesthetics. PMID- 11887374 TI - Emotional stress and brux-like activity of the masseter muscle in rats. AB - The aim of this study was to further clarify the relationship between emotional stress and bruxism. In experiment 1, 60 male 9-week-old Wistar rats were divided into four groups: the emotionally stressed (ES), the emotionally non-stressed (NS), the electrically foot-shocked (FSd), and the non-foot-shocked (NSd). ES rats were confined in a communication box for one hour a day to observe the emotional responses of neighbouring FSd rats. On days 0, 1, 4, 8, and 12, the electromyographic activity of the ES and NS rats' left masseter muscles was recorded for one hour, three hours after confinement in the communication box. Brux-like activity appeared in the masseter muscle of the ES group on days 1, 4, 8, and 12, but not in the NS group. In experiment 2, 36 male Wistar rats, 9 weeks old, were divided into three groups: emotionally stressed rats treated with an anti-anxiety drug (DES), emotionally stressed rats treated with saline as a vehicle (VES), and 24 FSd rats. Stress and EMG procedures were the same as those in experiment 1. Brux-like episodes decreased in DES rats from day 1 and significant differences were found on days 4 (P < 0.01), 8 (P < 0.05), and 12 (P < 0.05), when compared with the VES group. These findings suggest that emotional stress induces brux-like activity in the masseter muscle of rats, which was reduced with anti-anxiety drugs. PMID- 11887375 TI - An objective evaluation of the pitchfork analysis (PFA). PMID- 11887376 TI - Initial stress produced in the periodontal membrane by orthodontic loads in the presence of varying loss of alveolar bone: a three-dimensional finite element analysis. AB - The aim of this study was to investigate the stress components (S1 and S3) that appear in the periodontal membrane (PDM), when subjected to transverse and vertical loads equal to 1 N. A further aim was to quantify the alteration in stress that occurs as alveolar bone is reduced in height by 1, 2.5, 5, 6.5, and 8 mm, respectively. Six three-dimensional (3D) finite element models (FEM) of a human maxillary central incisor were designed. The models were of the same configuration except for the alveolar bone height. Special attention was paid to changes of the stress components produced at the cervical, apical, and sub-apical levels. In the absence of alveolar bone loss, a tipping force of 1 N produced stresses, which reached 0.072 N/mm2 at the cervical margin, up to 0.0395 N/mm2 at the apex and up to 0.026 N/mm2 sub-apically. In the presence of 8 mm of alveolar bone loss, the findings were -0.288, 0.472, and 0.722 N/mm2, respectively. Without bone loss, an intruding force of the same magnitude produced stresses of 0.0043, -0.0263, and 0.115 N/mm2, respectively, for the same areas and sampling points. In the presence of 8 mm of alveolar bone loss the findings were -0.019, 0.043, and 0.185 N/mm2 for intrusive movement. The results showed that alveolar bone loss caused increased stress production under the same load compared with healthy bone support (without alveolar bone resorption). Tipping movements resulted in an increased level of stress at the cervical margin of the PDM in all sampling points and at all stages of alveolar bone loss. These increased stress components were found to be at the sub-apical and apical levels for intrusive movement. PMID- 11887377 TI - Moisture-insensitive adhesives: reactivity with water and bond strength to wet and saliva-contaminated enamel. AB - The purpose of this study was to investigate the reactivity with water of a moisture-insensitive orthodontic primer (Transbond MIP), in conjunction with a no mix orthodontic adhesive (Unite), and a moisture-insensitive adhesive (Smartbond), and to assess their bond strength to wet and saliva-contaminated enamel relative to the conventional application of the no-mix adhesive. The reactivity of the moisture-insensitive products with water was investigated by micro-multiple internal reflectance Fourier transform infrared spectroscopy (micro-MIR FTIR). Sixty premolars were divided into two groups of 30 teeth each and, on the buccal enamel surfaces, a standardized volume (0.1 ml) of water or fresh whole saliva was applied. Standard edgewise stainless steel brackets were then bonded to enamel surfaces as follows: (a) Unite, (b) Unite with the Transbond MIP, and (c) Smartbond. The brackets were debonded under shear force at a speed of 2 mm/min and the debonded enamel surfaces were subjected to fractographic analysis. The statistical analysis of the bond strength values was performed by two-way ANOVA with condition (water, saliva) and adhesive type serving as discriminating variables (n = 10, alpha = 0.05). The results of the fractographic analysis were evaluated by chi 2 test (alpha = 0.05). FTIR analysis showed that only Smartbond set in the presence of water. Application of water in Transbond MIP increased the extent of carboxyl ionization without inducing any setting reaction. Transbond MIP did not improve bond strength values when combined with the no-mix adhesive. Most adhesive-enamel condition combinations showed a trend to present lower bond strength in the presence of saliva; however, this was not confirmed statistically. Fractography of enamel and bracket base surfaces showed that Unite + Transbond MIP resulted in the most adhesive fractures (leaving no resin on enamel surface), whereas Smartbond presented the highest frequency of cohesive fractures (adhesive left on bracket and enamel surfaces). PMID- 11887378 TI - Accuracy of linear and angular measurements on panoramic radiographs taken at various positions in vitro. AB - The accuracy of measurement of tooth length and angulation on dental panoramic tomograms (DPTs) is thought to be highly dependent on head positioning technique. A model representing the dentition and the functional occlusal plane was designed using an acrylic framework and stainless steel wires. The aim was to investigate whether varying the position of the model affects the linear and angular measurements on DPTs. Four different positions were investigated: initial position representing natural head posture (NHP) (T1); lateral right cant of the occlusal plane (T2); lateral left cant of the occlusal plane (T3); and tilting the occlusal plane up anteriorly (T4). On each DPT, four sets of measurements were recorded: (1) Vertical linear measurements of the stainless steel pins and ratio calculations of the 'crown' and 'root' segments (represented by the wire above and below the occlusal plane, respectively); (2) angular measurements of the pins relative to the occlusal plane; (3) angular measurements of the pins relative to a constructed reference line; and (4) angular measurements of pins relative to each other in the same segment. The results showed a significant error (P < 0.05) in all measurements when the occlusal plane was tilted up anteriorly by 8 degrees. A lateral cant of the occlusal plane by less than 10 degrees without an upward anterior rotation showed no significant effect on the measurements. This would suggest that there is some tolerance of variation in head position. PMID- 11887379 TI - The Aesthetic Component of the Index of Orthodontic Treatment Need validated against lay opinion. AB - This study aimed to determine the threshold of aesthetic impairment where orthodontic treatment would be sought by a sample of lay people. Using the 10 grade Aesthetic Component (AC) scale of the Index of Orthodontic Treatment Need (IOTN), 215 university students selected the level of aesthetic impairment that represented the point at which they would seek orthodontic treatment. Only nine (4.3 per cent) of the respondents recorded a threshold grade beyond grade 5 in the AC. The AC photograph grade 4 was found to be the most commonly selected threshold photograph. Subjects who visited the dentist every 6 months were more likely to choose a threshold photograph closer to the attractive end of the scale than those who visited their dentist less frequently (P < 0.01). This study, using lay people rather than dental health professionals, suggests that as currently used the AC does not reflect society's aesthetic expectations. The results indicate that when using the AC of the IOTN the 'no need for treatment' category should be grades 1-3 of the AC, rather than grades 1-4. PMID- 11887380 TI - Morphological differences in the craniofacial structure between Japanese and Caucasian girls with Class II Division 1 malocclusions. AB - The craniofacial features of 49 Japanese and 75 British Caucasian girls with Class II division 1 malocclusions were evaluated from lateral cephalometric radiographs, and the morphological differences between both races were examined. The subjects' ages ranged from 11 years 1 month to 12 years 11 months. The mean values of 13 linear and 13 angular cephalometric parameters were compared. The Japanese Class II division 1 sample had a significantly shorter anterior cranial base length (S-N; P < 0.001) and a more obtuse articular angle (S-Ar-Go; P < 0.001). Analysis of the dentoalveolar components in Japanese subjects showed more proclined lower incisors (L1/Go-Me; P < 0.05) and a steeper occlusal plane (Occ.P/S-N; P < 0.01) relative to those of Caucasians. The short anterior cranial base length and excessive vertical development in the Japanese population might be common racial morphological features, but the main reason for the Class II division 1 skeletal disharmony in both races was different; it was caused by the anteriorly positioned maxilla in Caucasians and the backward rotated mandible in the Japanese. PMID- 11887381 TI - Mandibular skeletal growth and modelling between 10 and 15 years of age. AB - This study pertains to a random sample of untreated French-Canadian adolescents (79 females and 107 males) evaluated at 10 and again at 15 years of age. Superimpositions on natural reference structures were performed to describe condylar growth and modelling of 11 mandibular landmarks. Superimpositions on natural cranial/cranial base reference structures were performed to describe mandibular displacement and true rotation. The results showed significant superior and posterior growth/modelling of the condyle and ramus. Males underwent significantly (P < 0.01) greater condylar growth and ramus modelling than females. With the exception of point B, which showed significant superior drift, modelling changes for the corpus landmarks were small and variable. The mandible rotated forward 2-3.3 degrees and was displaced 9.6-12.7 mm inferiorly and 1.9 2.7 mm anteriorly. Individual differences in ramus growth and modelling, both amount and direction, can be explained by mandibular rotation and displacements. Multivariate assessments revealed that superior condylar growth and ramus modelling were most closely associated with forward rotation and inferior mandibular displacement. Posterior growth and modelling were most closely correlated with anterior mandibular displacement and forward rotation. Modelling of the lower anterior border was independent of rotation and displacement. PMID- 11887382 TI - Intravenous sedation: an adjunct to enable orthodontic treatment for children with disabilities. AB - Intravenous (i.v.) sedation has become established as an important and preferred alternative to general anaesthesia (GA), in order to overcome difficulties in patient management encountered in the delivery of routine dental treatment of the disabled. However, its potential for the delivery of orthodontics for this group has never been exploited. The present pilot study describes the use of i.v. sedation with propofol to enable the performance of certain complex orthodontic and surgical procedures, which require strict control of the oral environment for prolonged periods, in a group of 10 of the most-difficult-to-manage disabled patients. The use of i.v. sedation provided a satisfactory management modality in these patients, who had previously been referred for GA. The parents reported complete satisfaction and general agreement that the same modality would again be welcomed. I.v. sedation significantly reduces the use of GA and makes treatment more readily available to a larger number of disabled patients. PMID- 11887383 TI - Cephalometric measurements and facial deformities in subjects with beta thalassaemia major. AB - This study was performed to identify cephalometric and facial features of patients with beta-thalassaemia major. A total of 54 thalassaemic subjects were examined for craniofacial deformities, including 37 patients (24 males and 13 females, aged 5-16 years) who had lateral cephalometric radiographs. The thalassaemic groups were compared with a normal control group matched for sex and dental age, using a t-test. All thalassaemic patients had a Class II skeletal base relationship. The average ANB angle was significantly larger than the controls in dental stages 2 and 3 (P < 0.05). Mandibular base length (Ar-Gn) was significantly less in thalassaemic patients than in controls, with the greatest differences (P < 0.001) found in the younger age group. The maxilla was of normal length (PNS-ANS, Ptm'-ANS') and appeared prominent (3.3 mm in males and 5.1 mm in females) due to a reduced cranial base length (Ar'-S') and a short mandible (Ar' P'). Vertically, thalassaemic patients showed a significantly increased maxillary/mandibular planes angle in all groups, with differences ranging between 6.19 and 12.55 degrees (P < 0.001). Thalassaemic patients also showed a reduced posterior facial height (S-Go, Ar-Go) and increased anterior facial proportions. Of the 54 thalassaemic patients examined, 17 per cent had severe facial disfigurements (grade 3). PMID- 11887384 TI - Changes in root length during orthodontic treatment: advantages for immature teeth. AB - The purpose of the study was to investigate root lengthening during orthodontic treatment in relation to the age of the patient, the developmental stage of the root, and the anticipated growth. Specifically, the potential benefit of treating young teeth was addressed. The sample consisted of 80 patients with Angle Class II division 1 malocclusions, treated with extraction of at least two maxillary first premolars, and edgewise technique with 0.018-inch slot brackets. Additionally, a cross-sectional control group of 66 untreated individuals matched to gender, and pre- and post-treatment age of the experimental group was included. Crown and root lengths of the maxillary incisors were measured on peri apical radiographs before and after treatment, and corrected for image distortion. The stage of root development before treatment was recorded. Root elongation during treatment was found for 50 out of the 280 examined teeth. Age at treatment start was significantly higher among the patients showing root shortening of the lateral incisors during treatment than among those showing root elongation (P < 0.05). The stage of root development was significantly related to the direction of root length change, i.e. shortening or elongation. Roots elongated during treatment did not differ in length from untreated teeth of similarly aged individuals. There was no significant difference in the extent of root lengthening between the roots elongated during treatment and the normal root lengthening in age-matched untreated individuals. Post-treatment root length was significantly related to pre-treatment age. Roots that were incompletely developed before treatment reached a significantly greater length than those that were fully developed at the start of treatment. The results of this study show a definite advantage for younger teeth with regard to post-treatment root length. This finding may influence treatment planning strategy. PMID- 11887385 TI - Fuzzy modelling for selecting headgear types. AB - The purpose of this study was to develop a computer-assisted inference model for selecting appropriate types of headgear appliance for orthodontic patients and to investigate its clinical versatility as a decision-making aid for inexperienced clinicians. Fuzzy rule bases were created for degrees of overjet, overbite, and mandibular plane angle variables, respectively, according to subjective criteria based on the clinical experience and knowledge of the authors. The rules were then transformed into membership functions and the geometric mean aggregation was performed to develop the inference model. The resultant fuzzy logic was then tested on 85 cases in which the patients had been diagnosed as requiring headgear appliances. Eight experienced orthodontists judged each of the cases, and decided if they 'agreed', 'accepted', or 'disagreed' with the recommendations of the computer system. Intra-examiner agreements were investigated using repeated judgements of a set of 30 orthodontic cases and the kappa statistic. All of the examiners exceeded a kappa score of 0.7, allowing them to participate in the test run of the validity of the proposed inference model. The examiners' agreement with the system's recommendations was evaluated statistically. The average satisfaction rate of the examiners was 95.6 per cent and, for 83 out of the 85 cases, 97.6 per cent. The majority of the examiners (i.e. six or more out of the eight) were satisfied with the recommendations of the system. Thus, the usefulness of the proposed inference logic was confirmed. PMID- 11887386 TI - The use of pressure garments on hypertrophic scars. AB - Hypertrophic scars are unsightly and can cause much distress to the sufferer. Pressure garments have been the preferred conservative management option for these scars for the last three decades. This paper reviews the history, underpinning theory and use of compression therapy in the management of hypertrophic scars particularly following burn injury. It is particularly aimed at clinicians who do not come into contact with this type of scarring frequently and are not familiar with its treatment. Other methods for managing hypertrophic scarring are also described briefly. Overall, there is a fair body of evidence to support the use of pressure garments, but it is not definitive research and further work is needed. However, the selection of any treatment must follow negotiation and agreement with the patient who will be required to continue treating their scars at home. PMID- 11887387 TI - A research review to identify the factors contributing to the development of pressure ulcers in paediatric patients. AB - The aim of this research review was to answer the clinical question 'what factors contribute to the development of pressure ulcers in the paediatric population?' A sensitive, systematic search was undertaken to identify all empirical studies that could answer this question. The question was broken down using facet analysis to identify the key terms that were used in the search strategy. All relevant databases were searched and a hand search of key journals was also undertaken. The review found very little empirical data that could answer the question. The majority of studies focused on the paediatric intensive care (PIC) population and had small sample sizes. Some factors were identified that contributed to the development of pressure ulcers in the PIC population. All of these factors except one were related to the acuity of the child's illness. The small sample sizes and limited populations mean that these studies cannot be generalised to the whole paediatric population and there is a need for more primary research to answer the question. PMID- 11887388 TI - Recording wound care effectiveness. AB - As more and more wound treatment regimes and wound care products become available, clinicians nowadays face the difficult task of choosing the right approach for individual patients amongst an ever expanding array of options. They find increasingly that a conventional subjective description of wound status and management approach in the patient's notes can no longer answer question such as how effective a treatment regime is or what the efficacy a selected wound care product may be at a given stage of the healing process. The only way to answer these questions and, ultimately, to improve patient care is the objective recording and documentation of the status of the wound and treatment related parameters. This paper outlines progress in a variety of technologies that are now beginning to become available in clinical practice, enabling practitioners to produce immediate, objective and quantifiable portraits of the healing process. PMID- 11887389 TI - Is there too much tissue viability? PMID- 11887390 TI - The experience and opinions of teachers of radiography students regarding pressure ulcer prevention and management in x-ray departments. AB - As part of a survey of pre-registration radiography course providers about pressure ulcer education, we asked respondents to comment on their personal experience of pressure ulcer prevention and management in x-ray environments, and also to identify what they consider to be important issues in this area. Responses were received from 14 out of the 24 providers of radiography education in the UK. The qualitative data was content analysed. The data suggests that radiography teachers have varied experience and a range of opinion about pressure ulcer prevention and management. The evidence supports partially anecdotal comments made to one of the authors (DJ) that radiographers cannot be expected to have knowledge of pressure ulcer prevention and management. Clinical staff with experience of pressure ulcer prevention and management should be aware of the potential risks in x-ray departments and provide support and education to radiography colleagues. PMID- 11887391 TI - Experimental copper deficiency, chromium deficiency and additional molybdenum supplementation in goats--pathological findings. AB - Secondary copper (Cu) deficiency, chromium (Cr) deficiency and molybdenosis (Mo) has been suggested to cause the "mysterious" moose disease in the southwest of Sweden. The present experiment was performed on goats to investigate the clinical, chemical, and pathological alterations after 20 months feeding of a semi-synthetic diet deficient in Cu and Cr. Four groups were included in the study: control group (n = 4), Cu-deficient group (group 1, n = 4), Cr-deficient group (group 2, n = 2) and Cu + Cr-deficient group (group 3, n = 3). Group 3 was additionally supplemented with tetrathiomolybdate during the last 2 months of the experiment. Main histopathological findings in groups 1 and 3 were the lesions in the liver, characterised by a severe active fibrosis, bile duct proliferation, haemosiderosis and mild necroses. Additionally, degenerative alterations of the exocrine pancreas were prominent in groups 1 and 3. Lesions in group 3 were more pronounced than in group 1. In group 3, the skin showed an atrophic dermatosis, while in group 2 a crusty dermatitis caused by Candida spp. was observed. This study shows that liver, pancreas and skin are mainly affected by a long term deficiency of copper and the findings are complicated by molybdenum application while chromium deficiency produced no histomorphological effects in our study. PMID- 11887392 TI - Antinuclear antibodies (ANA) in Gordon setters with symmetrical lupoid onychodystrophy and black hair follicular dysplasia. AB - Antinuclear antibodies (ANA) were demonstrated in 3 out of 10 Gordon setters with symmetrical lupoid onychodystrophy and in 5 out of 13 Gordon setters with black hair follicular dysplasia. Two dogs showed both symmetrical lupoid onychodystrophy and black hair follicular dysplasia, and one of these was ANA positive. The results suggest that symmetrical lupoid onychodystrophy and black hair follicular dysplasia in the Gordon setter might be autoimmune diseases that are pathogenetically related, which might indicate a common genetic predisposition. PMID- 11887393 TI - Serological investigation of granulocytic Ehrlichia infection in sheep in Norway. AB - Serum samples of 749 sheep from 75 sheep flocks in Norway, i.e. 361 lambs (6 to 7 months old) and 388 adults (> 1.5 year), were analysed for antibodies to Ehrlichia equi. Ten animals from each flock were examined. Seropositive animals were found along the coast of southern Norway from Vestfold to Sor-Trondelag (as far north as 63 degrees 38'N). Seropositive sheep were not found in southeast, east or northern Norway. Thirty-two flocks were seropositive, although tick-borne fever had only been diagnosed earlier in half of these. In 78% of the seropositive flocks, more than 80% of the sheep were seropositive. A total of 35.7% and 36.3% of lambs and adults were found seropositive, respectively. However, the overall seroprevalence among animals that had been grazing on Ixodes pastures were 0.80 for the lambs and 0.84 for the adults. Mean antibody titres (+/- SD) (log10) in seropositive lambs and adults were 2.59 (+/- 0.449) and 2.70 (+/- 0.481), respectively. No significant differences in either seroprevalence or mean antibody titre between sheep of different ages were obtained in this study. based on antibodies 94% of sheep flocks on Ixodes pastures were infected with a granulocytic Ehrlichia infection. The association between seropositive flocks and Ixodes infested pasture shows a very high degree of agreement (p < 0.00001). The present study indicates that granulocytic Ehrlichia infection in sheep is underdiagnosed in Norway. PMID- 11887395 TI - Persistence of granulocytic Ehrlichia infection during wintertime in two sheep flocks in Norway. AB - Granulocytic Ehrlichia infection in sheep is common in Norway in areas with Ixodes ricinus. In this study, 2 sheep flocks that had been grazing on I. ricinus infested pastures the previous season, were blood sampled after being housed indoors for nearly 6 months during wintertime. Thirty animals from each flock were examined for granulocytic Ehrlichia infection in the peripheral blood by blood inoculation studies, stained blood smear evaluation, polymerase chain reaction (PCR) analysis and serology (IFA-antibodies). The animals were sampled twice within a three-week period, the first time before and the second time after lambing. Two sheep in one flock were found Ehrlichia positive by both blood smear evaluation and PCR before lambing, and 3 sheep were found positive after lambing; 2 by blood smear examination and 3 by PCR. In the other flock, no sheep was found infected before lambing, but 2 ewes were found positive after lambing by both blood smear evaluation and PCR. In the first flock, 87% of the animals were found seropositive before lambing, and the mean antibody titre (log10 +/- SD) to E. equi was 2.45 +/- 0.401. In the second flock, 40% were found seropositive before lambing, and the mean antibody titre was 1.93 +/- 0.260. Seroprevalence and mean antibody titre in these 2 flocks were significantly different (p < 0.001). The present study indicates that sheep may be a reservoir host for granulocytic Ehrlichia infection from one grazing season to the next under natural conditions in Norway. PMID- 11887394 TI - The effect of two different oxytetracycline treatments in experimental Ehrlichia phagocytophila infected lambs. AB - The effect of 2 different oxytetracycline treatments in acute E. phagocytophila infected lambs was investigated. Twenty 5-month-old lambs of the Dala and Rygja breeds were used. Ten lambs were inoculated intravenously with a stabilate of an ovine E. phagocytophila strain. On the third day of fever, 4 lambs were given long-acting oxytetracycline (Terramycin prolongatum vet, Pfizer) (20 mg/kg) intramuscularly and another 4 lambs were given short-acting oxytetracycline (Terramycin vet, Pfizer) (10 mg/kg) intravenously for 5 consecutive days. The lambs were examined for the presence of Ehrlichia infection by blood smear evaluation, polymerase chain reaction (PCR) and antibody titre against E. equi. One month after the last antibiotic treatment, 250 ml citrate blood from each of these lambs were inoculated into each of 10 susceptible lambs, which were observed during the following 6 weeks. The results indicate that oxytetracycline given in the acute stage of the infection may effectively terminate the development of fever, rickettsemia and weight reduction in E. phagocytophila infected lambs. No difference was observed between the 2 treatment groups. However, at least 3 of 8 antibiotic treated lambs (37.5%) were still infected with granulocytic Ehrlichia 3 months after treatment. PMID- 11887396 TI - Regional eradication of Mycoplasma hyopneumoniae from pig herds and documentation of freedom of the disease. AB - The objectives of this study were to 1) screen all sow herds in a region for M. hyopneumoniae, 2) to effectuate an eradication programme in all those herds which were shown to be infected with M. hyopneumoniae, and 3) to follow the success of the screening and the eradication programmes. The ultimate goal was to eradicate M. hyopneumoniae from all member herds of a cooperative slaughterhouse (153 farrowing herds + 85 farrowing-to-finishing herds + 150 specialised finishing herds) before year 2000. During 1998 and 1999, a total of 5067 colostral whey and 755 serum samples (mean, 25 samples/herd) were collected from sow herds and analysed for antibodies to M. hyopneumoniae by ELISA. Antibodies were detected in 208 (3.6%) samples. Two farrowing herds (1.3%) and 20 farrowing-to-finishing herds (23.5%) were shown to be infected with M. hyopneumoniae. A programme to eradicate the infection from these herds was undertaken. During March 2000, a survey was made to prove the success of the screening and the eradication programmes. In total, 509 serum samples were collected randomly from slaughtered finishing pigs. Antibodies to M. hyopneumoniae were not detected in 506 of the samples, whereas 3 samples were considered suspicious or positive. Accordingly, 3 herds were shown to be infected. One of the herds was previously falsely classified as non-infected. Two of the herds were finishing herds practising continuous flow system (CF). Unlike finishing herds which practice all-in/all-out management routines on herd level, CF herds do not get rid of transmissible diseases spontaneously between batches, for which reason a screening was made in the rest of the CF herds (total n = 7). Consequently, 2 more infected herds were detected. In addition to the results of the survey, a decreasing prevalence of lung lesions at slaughter (from 5.2% to 0.1%) and lack of clinical breakdowns indicated that all member herds were finally free from M. hyopneumoniae in the end of year 2000. PMID- 11887397 TI - Diagnoses and treatments in health-classified fattening herds rearing pigs all in all out. AB - This study describes diseases encountered, medications used and veterinary involvement in all in--all out finishing herds belonging to one pork production system. The finishing herds had a particular management and housing regime. The pigs originated from health classified farrowing units. Information on 207,442 pigs was collected from 595 log books. Altogether 91% of the pigs received no treatments. Four percent of the batches of pigs were given antimicrobial mass medications. The local veterinarian visited the herds on average 2.6 times during the finishing period and made the diagnoses in more than half of the cases. At least one pig was affected with arthritis or tail biting in more than half of the batches, whereas locomotory diseases were recorded in one third of the batches. All other diagnoses were encountered in 1%-13% of the batches. Only a few pigs were treated individually in the affected groups. Antimicrobial drugs were given to 8% and other medicines to 0.7% of the pigs. The diagnosis was missing at least for one pig in 29% of the batches and the information about the medicine use in 8% of the treatments was missing. The study shows that it is possible to rear finishing pigs with only a small proportion of the animals needing treatments. The need of mass medications was low, because infectious diseases affecting the whole herd were uncommon. The recommendations for antimicrobial use given by the authorities had been followed quite well. The farmers and the veterinarians should be educated in order to realise the importance of proper record keeping. PMID- 11887398 TI - The dynamics, prevalence and impact of nematode infections in organically raised sheep in Sweden. AB - A three-year survey (1997-99) was carried out on organically reared sheep flocks throughout Sweden. The aim was to determine the prevalence and intensity of nematode infections and to establish relationships between sheep management practices and parasite infections. Faecal samples from ewes and lambs were collected from 152 organic flocks around lambing-time and during the grazing period for analysis. Results were compared with the different management practices that farmers use to prevent parasitism in their flocks. A high proportion of the flocks was infected with nematodes. The most prevalent species were Haemonchus contortus, Teladorsagia circumeincta, Trichostrongylus axei. T. colubriformis and Chabertia ovina and infections progressively increased during summer in lambs grazing on permanent pastures. Severity of parasitic infection in lambs was highly dependent on egg output from the ewes. H. contortus was found in 37% of the flocks, even at latitudes approximating the Polar Circle. Nematodirus battus was recorded for the first time in Sweden during the course of this study. Lambs turned out onto permanent pasture showed higher nematode faecal egg counts (epg) than lambs that had grazed on pastures, which had not carried sheep the previous year. This beneficial effect of lambs grazing non-infected pastures persisted if the ewes were treated with an anthelmintic before turn-out and if the lambs were kept on pastures of low infectivity after weaning. In lambs, the prevalence and the magnitude of their egg counts were higher during autumn in flocks where lambs were slaughtered after 8 months of age, compared with flocks where all lambs were slaughtered before this age. These results will be used in providing advice to farmers of ways to modify their flock management in order to minimise the use of anthelmintics, but at the same time efficiently produce prime lambs. PMID- 11887399 TI - Generation of EST and cDNA microarray resources for the study of bovine immunobiology. AB - Recent developments in expressed sequence tag (EST) and cDNA microarray technology have had a dramatic impact on the ability of scientists to study the responses of thousands of genes to external stimuli, such as infection, nutrient flux, and stress. To date however, these studies have largely been limited to human and rodent systems. Despite the tremendous potential benefit of EST and cDNA microarray technology to studies of complex problems in domestic animal species, a lack of integrated resources has precluded application of these technologies to domestic species. To address this problem, the Center for Animal Functional Genomics (CAFG) at Michigan State University has developed a normalized bovine total leukocyte (BOTL) cDNA library, generated EST clones from this library, and printed cDNA microarrays suitable for studying bovine immunobiology. Our data revealed that the normalization procedure successfully reduced highly abundant cDNA species while enhancing the relative percentage of clones representing rare transcripts. To date, a total of 932 EST sequences have been generated from this library (BOTL) and the sequence information plus BLAST results made available through a web-accessible database (http://gowhite.ans.msu.edu). Cluster analysis of the data indicates that a total of 842 unique cDNAs are present in this collection, reflecting a low redundancy rate of 9.7%. For creation of first generation cDNA microarrays, inserts from 720 unique clones in this library were amplified and microarrays were produced by spotting each insert or amplicon 3 times on glass slides in a 48-patch arrangement with 64 total spots (including blanks and positive controls) per patch. To test our BOTL microarray, we compared gene expression patterns of concanavalin A stimulated and unstimulated peripheral blood mononuclear cells (PBMCs). In total, hybridization signals on over 90 amplicons showed upregulation (> 3x) in response to Con A stimulation, relative to unstimulated cells. A second experiment with PBMCs from a different group of animals was performed to test reproducibility of microarray results. There was a high correlation between the 2 experiments (r = 0.72, P < 0.001). Resources described in this publication offer a highly efficient and integrated system to study gene expression changes in bovine leukocytes. PMID- 11887400 TI - An immunogenomics approach to understanding periparturient immunosuppression and mastitis susceptibility in dairy cows. AB - Studies comparing in vivo and in vitro functional capacities of leukocytes from non-parturient and periparturient dairy cows have provided substantial evidence that systemic and local mammary immune defenses are deficient around parturition. This evidence has lead to the reasonable hypothesis that immune deficiency underlies the heightened mastitis susceptibility of periparturient cows. Nutrition and vaccine studies substantiate this hypothesis, showing that dietary antioxidant supplementation and rigorous immunization regimes can bolster innate and humoral immunity to the point that mastitis severity and time for return to normal milk production are reduced. However, completely effective resolution of this significant production disease has not been achieved because so little is understood about its complex etiology. In particular, we possess almost no knowledge of how or why immune cells responding to parturient physiology end up with deficient functional capacities. Fluctuations in reproductive steroid hormones and chronic shifts in neuroendocrine hormones with roles in nutrient partitioning and appetite control may affect the expression of critical leukocyte genes in periparturient dairy cows. A thorough understanding of leukocyte biology during periparturition would seem a critical goal for future development of effective mastitis prevention strategies. Recently, our group has begun to use cDNA microarray technology to explore bovine leukocyte RNA for global gene expression changes occurring around parturition. We are working within the context of a hypothesis that the physiology of parturition negatively affects expression of critical genes in blood leukocytes. In the current study we initiated hypothesis testing using leukocyte RNA from a high producing Holstein cow collected at 14 days prepartum and 6 hours postpartum to interrogate a cDNA microarray spotted with > 700 cDNAs representing unique bovine leukocyte genes. This analysis revealed 18 genes with > or = 1.2-fold higher expression 14 days prepartum than 6 hours postpartum. BLASTN analysis of these genes revealed only one that can be considered a classical immune response gene. All other repressed genes were either unknown or putatively identified as encoding key proteins involved in normal growth and metabolism of cells. Given the critical roles of these repressed genes in normal cell functions, we may have identified good candidates to pursue with respect to periparturient immunosuppression and mastitis susceptibility. PMID- 11887401 TI - An unusual congenital malformation in a calf with serological evidence of foetal bovine viral diarrhoea virus infection. PMID- 11887402 TI - HIPAA compliance date for electronic transactions extended. PMID- 11887403 TI - Drug products not to be exposed to postal irradiation. PMID- 11887404 TI - California pharmacists to offer emergency contraceptive services. PMID- 11887405 TI - Drug industry sues states over prescription programs for low-income residents. PMID- 11887406 TI - Cefditoren pivoxil. PMID- 11887407 TI - Hawthorn: pharmacology and therapeutic uses. AB - The uses, pharmacology, clinical efficacy, dosage and administration, adverse effects, and drug interactions of hawthorn are discussed. Hawthorn (Crataegus oxyacantha) is a fruit-bearing shrub with a long history as a medicinal substance. Uses have included the treatment of digestive ailments, dyspnea, kidney stones, and cardiovascular disorders. Today, hawthorn is used primarily for various cardiovascular conditions. The cardiovascular effects are believed to be the result of positive inotropic activity, ability to increase the integrity of the blood vessel wall and improve coronary blood flow, and positive effects on oxygen utilization. Flavonoids are postulated to account for these effects. Hawthorn has shown promise in the treatment of New York Heart Association (NYHA) functional class II congestive heart failure (CHF) in both uncontrolled and controlled clinical trials. There are also suggestions of a beneficial effect on blood lipids. Trials to establish an antiarrhythmic effect in humans have not been conducted. The recommended daily dose of hawthorn is 160-900 mg of a native water-ethanol extract of the leaves or flowers (equivalent to 30-169 mg of epicatechin or 3.5-19.8 mg of flavonoids) administered in two or three doses. At therapeutic dosages, hawthorn may cause a mild rash, headache, sweating, dizziness, palpitations, sleepiness, agitation, and gastrointestinal symptoms. Hawthorn may interact with vasodilating medications and may potentiate or inhibit the actions of drugs used for heart failure, hypertension, angina, and arrhythmias. The limited data about hawthorn suggest that it may be useful in the treatment of NYHA functional class II CHF. PMID- 11887408 TI - Interdisciplinary medication education in a church environment. AB - A medication education program for ambulatory care patients implemented in a church setting was studied. The program at each of 20 churches in Ohio consisted of a one-hour orientation for pharmacists/interns, a 20-minute presentation on medications and health, a question-and-answer session led by a pharmacist and a nurse, a one-on-one session with a pharmacist, and an exit interview with a nurse. Before the program, patients completed a form to assess their current experiences with medications and their interactions with health care professionals in the preceding six months. During an exit interview at the end of the program, patients were asked whether the program has been understandable and beneficial and whether taking medications affected their lifestyle. A follow-up interview was conducted six months later. A total of 187 patients completed both the exit and follow-up interviews. Almost all reported that the church setting was a good place for the program and that the program was beneficial. During the six months after the program, the patients took significantly fewer drugs each day than during the six months before the program and had fewer drug-related problems. Significantly more patients sought drug information after completing the program than before it. High rates of medication misuse were identified, leading to 359 pharmacist recommendations. An interdisciplinary program in a church setting successfully provided medication education. PMID- 11887409 TI - Current management of anemia in adult hemodialysis patients with end-stage renal disease. AB - The management of anemia in adult end-stage renal disease (ESRD) patients receiving hemodialysis in dialysis facilities is examined. Clinical information was collected for a random sample of adult (age > or = 18 years) patients who received hemodialysis for ESRD between October and December 1999 and included hemoglobin concentrations, epoetin alfa doses and routes of administration, iron prescribing patterns, transferrin saturation levels, and serum ferritin concentrations. Patients whose data did not include hemoglobin concentrations with the weekly epoetin dose were excluded from the analysis. Associations by patient characteristics and geographic region were examined for clinical intermediate outcomes and epoetin alfa and iron prescribing practice patterns. Data were submitted for 8154 patients, and hemoglobin values linked to weekly epoetin alfa doses were available for 7573 of those patients. The mean hemoglobin concentration for patients in the sample was 11.4 +/- 1.3 g/dL. Sixty-seven percent of patients had mean hemoglobin values > or = 11 g/dL. Females, blacks, patients 18-44 years old, and patients receiving hemodialysis for less than six months exhibited significantly lower mean hemoglobin values despite being prescribed, on average, significantly higher epoetin alfa doses than males, whites, older patients, and patients receiving hemodialysis for six months or more (p < 0.001). There was significant regional variation in the prescribing patterns for s.c. epoetin alfa and i.v. iron (p < 0.001). Multivariable logistic regression analysis found significant associations between mean hemoglobin values > 11 g/dL and certain patient characteristics, including white race, hemodialysis for six months or longer, lower prescribed weekly epoetin alfa doses, prescription of i.v. iron, mean transferrin saturation levels > or = 20%, mean Kt/V > or = 1.2, and higher mean serum albumin values. Prescribing patterns for i.v. iron did not vary by the status of patients' iron stores. Regional and patient-specific variations in parameters of anemia management provide pharmacists with the opportunity to contribute to a multidisciplinary team approach to improve the care of hemodialysis patients. PMID- 11887411 TI - Role of clinical pharmacists in outpatient HIV clinics. PMID- 11887410 TI - Comparison of methods for detecting medication errors in 36 hospitals and skilled nursing facilities. AB - The validity and cost-effectiveness of three methods for detecting medication errors were examined. A stratified random sample of 36 hospitals and skilled nursing facilities in Colorado and Georgia was selected. Medication administration errors were detected by registered nurses (R.N.s), licensed practical nurses (L.P.N.s), and pharmacy technicians from these facilities using three methods: incident report review, chart review, and direct observation. Each dose evaluated was compared with the prescriber's order. Deviations were considered errors. Efficiency was measured by the time spent evaluating each dose. A pharmacist performed an independent determination of errors to assess the accuracy of each data collector. Clinical significance was judged by a panel of physicians. Observers detected 300 of 457 pharmacist-confirmed errors made on 2556 doses (11.7% error rate) compared with 17 errors detected by chart reviewers (0.7% error rate), and 1 error detected by incident report review (0.04% error rate). All errors detected involved the same 2556 doses. All chart reviewers and 7 of 10 observers achieved at least good comparability with the pharmacist's results. The mean cost of error detection per dose was $4.82 for direct observation and $0.63 for chart review. The technician was the least expensive observer at $2.87 per dose evaluated. R.N.s were the least expensive chart reviewers at $0.50 per dose. Of 457 errors, 35 (8%) were deemed potentially clinically significant; 71% of these were detected by direct observation. Direct observation was more efficient and accurate than reviewing charts and incident reports in detecting medication errors. Pharmacy technicians were more efficient and accurate than R.N.s and L.P.N.s in collecting data about medication errors. PMID- 11887412 TI - Physical compatibility of milrinone lactate injection with intravenous drugs commonly used in the pediatric intensive care unit. PMID- 11887413 TI - Judicial policy and expanded duties for pharmacists. PMID- 11887414 TI - Visual compatibility of abciximab with selected drugs. PMID- 11887415 TI - Oxcarbazepine and hyponatremia. PMID- 11887416 TI - Beta-adrenergic blockers in chronic congestive heart failure. PMID- 11887417 TI - Detection of human rotavirus in faeces from diarrhoeic calves in north-east Nigeria. AB - Information on the epidemiology of rotavirus in any particular area is necessary for vaccine development against the disease caused by the virus. This study presents preliminary information on the prevalence of human rotavirus in diarrhoeic calves in North-east Nigeria. Faecal samples from 188 diarrhoeic calves in various farms in North-east Nigeria, obtained between November 1998 and February 1999, were analysed by ELISA for the presence of rotaviruses. A prevalence rate of 3.2% was recorded, with the virus being prevalent among calves aged 29-56 days (p < 0.05). The implications of these findings are discussed with respect to the close association between the herdsmen and their animals and the sharing of a common source of drinking water in the predominantly livestock producing communities of North-east Nigeria. PMID- 11887418 TI - Prevalence and aetiology of mastitis in cows from two major Ethiopian dairies. AB - The study was undertaken to determine the aetiology and prevalence of mastitis in hand-milked cows (n = 186) in two major Ethiopian dairies. The California Mastitis Test and culturing for bacteria revealed that 21.5% of the cows were clinically infected and 38.2% had subclinical mastitis. Most mastitis pathogens isolated from milk samples testing positive by the California Mastitis Test were Gram-positive cocci. Staphylococci constituted 57% of the isolates, of which the predominant cause of bovine mastitis was Staphylococcus aureus (40.5%). Other mastitis pathogens isolated include streptococci (16.5%), coliforms (9%) and corynebacteria (5%). Retrospective analysis of farm records indicated that mastitis was the second most important cause of culling and accounted for 27% of the cows removed from these two dairies. PMID- 11887419 TI - Efficacy of sex determination in the greater cane rat, Thryonomys swinderianus, Temminck. AB - The efficacy of the two most common techniques used for determining the sex of the greater cane rat, Thryonomys swinderianus, Temminck, was tested using 10 young and 8 adult animals with two technicians at the Animal Research Institute's Grasscutter Domestication Centre, Pokoase, Ghana. The techniques compared were the use of the head shape and/or head size and the use of the ano-genital distance. The use of the ano-genital distance for sex determination was validated in a colony of greater cane rats at various stages of development, i.e. from the day of birth to three or more years of age. The ano-genital distance was then used as the standard against which the use of the head shape and/or head size technique was tested. The results indicated that the use of the ano-genital distance for sex determination in the greater cane rat was error-free, even with little experience on the part of the practitioner. The ano-genital distance was more than twice as long in the males than in the females at all ages (p < 0.001). However, the use of the head shape and/or head size for sex determination was found to be associated with some degree of error in both sexes. The use of the ano-genital distance can therefore be recommended as an efficacious technique for sex determination in the greater cane rat. PMID- 11887420 TI - Detection of oestrus in Bunaji cows under field conditions. AB - The objective of this study was to determine the efficacy of some aids for detecting oestrus in 72 Bunaji cows synchronized using two injections of prostaglandin F2 alpha (PGF2 alpha) 13 days apart. Blood samples to determine the accuracy of the aids and ovarian activity by radioimmunoassay technique were collected daily from the day of the second PGF2 alpha injection until oestrus or for 168 hours for 'non-responders'. The aids for detecting oestrus, tail painting, KaMar detectors and a chin-ball mating device (CMD) were applied at the time of the second injection. The oestrus response rate was 73.6% and 61.1%, from the serum progesterone concentration and visual observation, respectively. Based on the total number of oestrus events observed, unaided visual observation, tail painting, KaMar and CMD detected 52.2%, 82.6%, 82.6% and 76.8%, respectively. Visual observation of standing oestrus alone failed to detect 47.8% of the occurrence of oestrus as shown by the concentration of progesterone in the serum and 30.4%, 30.4% and 24.6%, respectively, of the oestrus periods recorded by tail painting, KaMar and CMD. Twenty-eight cows showed abnormalities in progesterone concentration. These investigations showed that oestrus may be detected by using the aids in cows that have been regarded as 'anoestrous' by visual observation of standing heat. PMID- 11887421 TI - Effect of cowpea hay, groundnut hay, cotton seed meal and maize meal supplementation to maize stover on intake, digestibility, microbial protein supply and acetate kinetics in weaner lambs. AB - Ten weaner lambs were used in a double 5 x 5 Latin square design to evaluate the effect of supplementing maize stover (MS) with cowpea hay (CW), groundnut hay (GN), cotton seed meal (CSM) or maize meal (MM) on the intake, digestion kinetics and acetate clearance rate. CW and GN were offered at 30% w/w to MS, while CSM and MM were given at 15 g/kg0.75 per day. Supplementation reduced (p < 0.01) MS intake but enhanced (p < 0.01) total dry matter intake. There were no significant differences (p > 0.05) in digestibility. However, the estimated ME intake was significantly (p < 0.05) improved by supplementation. The estimated microbial protein supply was almost significantly (p < 0.06) improved by 22.68%, 5.35%, 17.58% and 47.90% on the CW-, GN-, CSM- and MM-supplemented diets, compared to the control (7.85 g/day). Microbial protein synthesis efficiency was not significantly affected (p > 0.05) by diet, and nor were the acetate clearance rates (p > 0.05), which averaged 0.0475 +/- 0.0078/min. The improvement in ME intake may have been due to a faster flow rate of digesta and a better balance of nutrients in the end-products of digestion. These results demonstrate that small amounts of forage supplements can improve nutrient intake when animals consume low-quality forages and provide a basis for comparing such supplements with bought-in protein and energy supplements. PMID- 11887422 TI - Effect of supplementation with leguminous crop residues or concentrates on the voluntary intake and performance of Kirdi sheep. AB - Twenty young Kirdi (West African Dwarf) rams, averaging 22 kg live weight and 15 months of age, were randomly assigned to four feeding groups of 5 animals each. The groups were subjected to an 8-week feeding trial to determine the effect of different protein supplements on their voluntary intake of rice straw and on their performance. The animals in one of the groups were maintained on a daily basal diet of rice straw given ad libitum and 250 g of rice bran per animal (control). Other groups were fed the basal diet, supplemented with 300 g of groundnut haulms (GH diet), 45 g of cotton seed cake (CSC diet) or 210 g of chopped cowpea vines (CPV diet) per animal per day. The average daily weight gains of the animals feeding on the control, GH, CSC and CPV diets were 20.00, 48.93, 52.14 and 49.29 g, respectively. The gains in live weight of the supplemented groups did not differ significantly (p > 0.05), but there was a difference (p < 0.05) in live weight gain between the supplemented groups and the control. The differences in intake of rice straw among the groups were highly significant (p < 0.01). Cotton seed cake increased (p < 0.01) the intake, while feeding crop residues tended to depress the intake of rice straw. Variations in the height at withers, heart girth and scapulo-ischial length did not differ significantly (p > 0.05) among the groups. In terms of live weight gain and intake of rice straw, cotton seed cake was the most effective supplement, but, since the crop residues are more readily available to farmers in this area of the country, they merit more attention. PMID- 11887423 TI - Purification and characterization of the aetiological agent of hydropericardium hepatitis syndrome from infected liver tissues of broiler chickens. AB - Hydropericardium hepatitis syndrome in broiler chickens is an acute, infectious disease characterized by high mortality, excess pericardial fluid and multifocal hepatic necrosis. The aetiological agent was purified to homogeneity from infected liver tissues from field outbreaks. Electron-microscopic and serological confirmation of the virus were undertaken and the disease was reproduced experimentally in broiler chicks. The results indicated that an adenovirus, fowl adenovirus serotype 4, was alone responsible for the disease in the materials studied. PMID- 11887424 TI - A possible association between dietary intake of copper, zinc and phosphate and delayed puberty in heifers in Sudan. AB - Zinc and copper deficiencies have been reported in heifers of various breeds at four different locations in Sudan. These were Kuku (5 km north of Khartoum), Seleit (20 km northwest of Khartoum), Medani (180 km south of Khartoum) and El Obeid (600 km west of Khartoum). Phosphorus deficiency was only observed in the serum of heifers at El Obeid. The heifers at all locations showed delayed puberty, stunted growth and infertility. The heifers of the local breeds at El Obeid only attained puberty by 1530 days of age compared with 840 days for the pure Friesian heifers at Seleit. The crossbred animals at Kuku and Medani attained puberty at 1440 and 1020 days of age, respectively. The marginal or low zinc and copper contents in pasture, soil or animal feed may have been predisposing factors for the observed deficiencies and might have been responsible for the delayed age of puberty. PMID- 11887425 TI - Changes in blood glucose, plasma non-esterified fatty acids and insulin in pregnant and non-pregnant goats. AB - The blood glucose and the plasma non-esterified fatty acids (NEFA) and insulin concentrations were estimated in jugular blood samples from 18 Alpine x Beetal and Sannen x Beetal goats during pregnancy and compared with samples from non pregnant goats and from goats during the periparturient period. The blood glucose levels in the pregnant goats rose to a peak of about 60 +/- 1.36 mg/ml at 42-56 days and then declined to about 46 +/- 2.37 mg/ml at 112-126 days. In non pregnant goats, the blood glucose levels were significantly (p < 0.01) higher than in pregnant goats, except between days 42 and 70 (59 +/- 1.36 mg/ml). On the day of kidding, the levels declined significantly (p < 0.01), increasing again thereafter. The plasma NEFA concentrations were significantly higher in pregnant than in non-pregnant goats from days 56 to 126. The NEFA concentration increased on the day of kidding, followed by a transient fall by day 3. The plasma insulin concentration was usually higher in pregnant than in non-pregnant goats, except between days 56 and 70 and from day 126 onwards. The insulin concentration fell late in pregnancy, but there was a transient increase 2 days after parturition. The blood glucose and plasma NEFA concentrations can be used as indices of nutritional status during pregnancy in goats. PMID- 11887426 TI - Factors associated with changes in serum total cholesterol levels over 7 years in middle-aged New Zealand men and women: a prospective study. AB - BACKGROUND AND AIM: The aim of this study was to determine the factors associated with changes in serum total cholesterol levels over a period of seven years. METHODS AND RESULTS: The baseline Workforce Diabetes Survey was carried out between 1988 and 1990 and involved workers predominantly aged > or = 40 years; a follow-up survey of 4,053 participants was carried out between 1995 and 1997. Both surveys measured serum lipid levels by means of enzymatic methods. The overall age- and gender-adjusted mean serum cholesterol levels decreased by 4.6% between the two surveys. The two-thirds of participants who experienced a decrease in total serum cholesterol formed a higher risk group at baseline insofar as they were older, more inactive and more likely to be male, and had higher blood pressure (BP), higher serum cholesterol and triglyceride levels, and a higher body mass index (BMI) and waist/hip ratio than the one-third whose serum cholesterol levels increased (all p < 0.05). The decrease in serum cholesterol was associated with improvements or less deterioration in risk factors (fasting glucose, BP, BMI and the waist/hip ratio, the low-density/high-density lipoprotein (LDL/HDL) ratio, triglyceride concentrations and level of physical activity) and an increase in the use of lipid lowering drugs. CONCLUSION: Serum cholesterol levels decreased over the seven years between the surveys, principally among the individuals at highest risk. The use of lipid lowering drugs contributed to this decline but lifestyle factors, such as increased exercise levels, may also have played a role because other risk factors also improved. PMID- 11887427 TI - Insulin resistance, ventricular mass and function in normoglycaemic hypertensives. AB - BACKGROUND AND AIM: To investigate the possible relationship between insulin sensitivity and the main non-invasively determined parameters of left ventricular structure and function in an early phase of hypertensive disease. METHODS AND RESULTS: We studied a group of 60 patients with previously untreated primary arterial hypertension of recent onset. Insulin sensitivity was evaluated by means of homeostasis model assessment (HOMA), and cardiac structure and function by means of ultrasonography. The patients were subsequently divided on the basis of HOMA tertiles and the between-group differences were statistically evaluated using one-way analysis of variance. No difference was found in the main demographic parameters, but high-density lipoprotein (HDL)-cholesterol was significantly higher (1.43 +/- 0.4, 1.16 +/- 0.2, 1.16 +/- 0.2 mmol/L; p < 0.05) and total triglycerides significantly lower (0.94 +/- 0.3, 1.41 +/- 0.8, 1.54 +/- 0.7 mmol/L; p < 0.05) in the first tertile. In relation to the non-invasive parameters of diastolic function, the E wave was significantly reduced in the third tertile (0.64 +/- 0.1, 0.63 +/- 0.1, 0.52 +/- 0.1 m/s; p < 0.05), with a concomitant significant increase in AFF (0.51 +/- 0.03, 0.51 +/- 0.06, 0.56 +/- 0.03; p < 0.05). The E wave (r = 0.326, p < 0.05), E/A wave ratio (r = -0.277, p < 0.05) and AFF (r = 0.432, p < 0.001) were significantly related to HOMA, regardless of age, body mass index, heart rate, left ventricular mass and blood pressure. CONCLUSIONS: Our results indicate that hypertensive patients in the third HOMA tertile (i.e. those with reduced insulin sensitivity) have early diastolic left ventricular abnormalities. PMID- 11887428 TI - Left ventricular geometry and arterial function in hypercholesterolemia. AB - AIMS: To investigate the effect of hypercholesterolemia on total arterial compliance and left ventricular (LV) geometry in the absence of arterial hypertension and diabetes. METHODS: One hundred and fifty-two normotensive, non diabetic patients (109 men) aged 52 +/- 10 years with plasma cholesterol > 240 mg/100 mL, and 282 normotensive controls (154 men) aged 42 +/- 10 years (p < 0.0001) with plasma cholesterol < 200 mg/100 mL were studied by means of echocardiography. The stroke volume/pulse pressure ratio as a percentage of the value predicted by individual age, body weight and heart rate was used as a prognostically-validated index of total arterial compliance. Central pulse pressure (PP) was estimated using a regression equation obtained in a non overlapping population. RESULTS: Although within the "normal" range, systolic pressure, PP and estimated central PP were higher in the hypercholesterolemic patients even after controlling for differences in age, body mass index (BMI) and race (all p < 0.0001). After controlling for differences in systolic pressure, age, BMI and race, LV mass and the prevalence of hypertrophy were comparable between the two groups, whereas relative diastolic wall thickness was greater (0.36 + 0.06 vs 0.33 + 0.05) and percent SV/PP (stroke volume/PP) lower in the hypercholesterolemic patients (96 +/- 19% vs 102 +/- 18%; both p < 0.005). After considering the covariates, there was still an independent negative correlation between relative wall thickness and percent SV/PP (r = -0.37, p < 0.0001). CONCLUSIONS: Hypercholesterolemia in normotensive non-diabetic adults is independently associated with a mildly concentric LV geometry and a reduced index of total arterial compliance. PMID- 11887429 TI - Serum cholesterol response to replacing butter with a new trans-free margarine in hypercholesterolemic subjects. AB - Margarine leads to lower total and LDL cholesterol (LDL-C) levels than butter but may contain trans fatty acids that increase atherogenic lipids. A food company has used data concerning the cholesterolemic effects of individual fatty acids, including trans fatty acids, to develop a commercially available and virtually trans-free margarine. OBJECTIVE: The effect of this novel margarine on serum lipids and lipoproteins was compared with that of butter in free-living, hypercholesterolemic subjects. DESIGN AND SETTING: A two-period, outpatient cross over trial at a university hospital lipid clinic. SUBJECTS: The study involved 77 subjects, and was completed by 53 men and 19 women aged 35-65 years with total serum cholesterol levels of between 6.0 and 7.9 mmol/L. INTERVENTION: Two 23-day regimens, separated by a 4-week washout period, included individualised dietary prescriptions supplemented with butter or margarine designed to provide 15% of total dietary energy. RESULTS: In comparison with butter, margarine intake lowered total and LDL-C levels by respectively 11.1% (99% CI: 8.1-14.1) and 11.3% (99% CI: 7.6-15.1). The reduction in LDL-C was < 3% in nearly one-fifth of the subjects despite appropriate changes in serum triglyceride fatty acids. Of the tested clinical and demographic variables, only the percentage of energy obtained from saturated fat during the margarine intake period was associated with dietary responsiveness (explaining 12% of the variation; p < 0.01). CONCLUSION: Our results suggest that a margarine designed to meet nutritional recommendations for hypercholesterolemia is more efficacious than butter in reducing atherogenic lipid levels in hypercholesterolemic subjects. PMID- 11887430 TI - Clinically relevant pleiotropic effects of statins: drug properties or effects of profound cholesterol reduction? AB - Clinical trials have firmly established that 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) can induce the regression of vascular atherosclerosis and reduce cardiovascular-related morbidity and death in patients with and without coronary artery disease. It is usually assumed that these beneficial effects are due to the ability of statins to reduce cholesterol synthesis. However, because mevalonic acid is not only the precursor of cholesterol but also of many non-steroidal isoprenoid compounds, the inhibition of HMG-CoA reductase may lead to pleiotropic effects. As shown by the data reported in this review, some statins can interfere with major events involved in the formation of atherosclerotic lesions, regardless of their hypolipidemic properties. The relevance of these effects in humans remains to be established (particularly in view of the high statin doses required to produce a direct vascular action), thus their contribution to the reduction in cardiovascular events observed in clinical trials has become one of the major challenges for future studies aimed at clarifying the anti-atherosclerotic benefits of statins. PMID- 11887431 TI - Hypertension in obesity. AB - AIM: To review various topics regarding the relationship between obesity and hypertension. DATA SUMMARY: Obesity is a widespread and increasingly prevalent condition associated with a large number of comorbid diseases, one of the most important of which is obesity-induced hypertension (HTN). The association between obesity and HTN has been well documented in most racial, ethnic and socio economic groups, although the relationship between body mass index (BMI) and blood pressure values depends on age, gender, type of obesity and race differences. Obesity-induced HTN has some unique characteristics that differ from those observed in lean hypertensive patients. The hemodynamic profile of obese subjects is characterised by high cardiac output, high plasma and total blood volume, and inappropriately normal to total peripheral resistance. Clinically, hypertensive obese subjects are more likely to develop left ventricular hypertrophy and kidney damage than their lean counterparts. Various common factors are involved in establishing sodium retention and vascular resistance and may be critically influenced by the neurobiological/genetic mechanisms leading to obesity, in which insulin, leptin and the adrenergic system play major roles. Obesity is one of the main causes of therapeutic failure, and a number of studies have demonstrated that obese subjects need more antihypertensive drugs than sex and age-matched lean hypertensives. Long-term dietary treatment, consisting of a moderate restriction of energy and salt intake, is the most effective and safe treatment for obesity-associated HTN. The use of treatments other than calorie restriction should be considered with caution. Drugs that increase energy expenditure or reduce appetite may variably increase blood pressure (BP) and are unsuitable for hypertensive subjects. There do not seem to be any clear differences in the efficacy of the various antihypertensive drug classes. The clustering of cardiovascular risk factors other than HTN needs to be taken into account when choosing antihypertensive treatment for obese subjects. CONCLUSIONS: Obesity is a highly prevalent condition that causes or exacerbates many health problems including HTN. Combined interventions at different levels can help in losing weight and therefore reduce the cardiovascular risk, morbidity and mortality associated with obesity. PMID- 11887432 TI - Impact of obesity in primary hyperlipidemias. AB - Obesity is frequently associated with high plasma triglyceride and reduced plasma high-density lipoprotein (HDL)-cholesterol (HDL-C) levels, and an increased concentration of apoB-carrying lipoproteins. The effects of obesity on lipid metabolism are mainly mediated by insulin resistance and, as central (visceral) obesity significantly increases insulin resistance, it aggravates these lipid changes. We have reviewed the impact of obesity on lipid metabolism in different types of primary hyperlipidemias. Obesity is not common in primary (familial and polygenic) hypercholesterolemias, and insulin resistance is infrequent; various investigators have found no or only a weak association between plasma cholesterol concentrations and insulin levels. On the other hand, in familial hypertriglyceridemia (type IV) and familial combined hyperlipidemia (FCH), obesity and insulin resistance are common and, when present, contribute to a further deterioration in the lipid profile. Weight loss in most of these patients is accompanied by a significant decrease in plasma triglyceride levels and an increase in HDL-C. Reviewing the data published by our group, we show that insulin resistance is an important component of the metabolic derangement in FCH subjects; high fasting plasma free fatty acids and triglycerides levels correlate to insulin resistance, thus linking this abnormality to lipid metabolism. A high waist/hip ratio (indicating visceral fat deposits) exacerbates insulin resistance, but this is also present in lean FCH subjects. Furthermore, insulin resistance is associated with a higher prevalence of coronary heart disease in this group of subjects. PMID- 11887433 TI - [Pathogenesis of the progression of bronchial lesions in bronchiectasis]. PMID- 11887434 TI - [Diagnosis, etiology, course of bronchiectasis in adults: apropos of a retrospective study conducted at the CHU of Rennes concerning 127 patients hospitalized between 1992 and 1994]. PMID- 11887435 TI - [Antibiotic strategy in bronchiectasis in children (mucoviscidosis excepted)]. PMID- 11887436 TI - [Antibiotic strategies in bronchiectasis of adults]. PMID- 11887437 TI - [Management of bronchiectasis antibiotic therapy excepted. Non-antibiotic therapy of children and adults]. PMID- 11887438 TI - [Diagnosis, etiology, course of bronchiectasis in children]. PMID- 11887439 TI - Piercing and tooth jewelry--an ethical dilemma. PMID- 11887440 TI - Refusal for general anaesthetic. PMID- 11887441 TI - Infection control among dentists in private practice in Durban. AB - The general fear, superstition and alarm surrounding HIV/AIDS warrant that the highest standards of care be available to our patients. A survey on infection control was undertaken in Durban to assess the current state of infection control procedures among dentists in private practice. A self-administered 44-item questionnaire was hand-delivered to a random sample of 75 dentists (31.3%)--see comments in Methods--in private practice. The response rate was 90.7% (68 dentists). The routine use of gloves, masks, and protective eyewear was reported by 97.1%, 82.4% and 52.9% of dentists respectively. Although 89.7% of dentists had autoclaves in their practices, only 45.2% autoclaved their high speed handpieces and 39.7% their slow handpieces. Almost 60% of dentists did not use rubber dam at all whilst 46.3% did not disinfect impressions before sending them to the laboratory. Approximately 6% of respondents reported re-using local anaesthetic cartridges and 1.5% re-used needles. Needlestick injuries in the previous six months were reported by 13.8% of dentists but two thirds of them did not follow any specific protocol after injury. Almost 90 per cent of dentists were immunised against Hepatitis B but more than 60% of their staff were not. The results of the study showed that adherence to universally accepted guidelines for infection control remain low amid a climate of an ever-increasing HIV pandemic. PMID- 11887442 TI - Infection control in South African oral hygiene practice. AB - OBJECTIVE: The aim was to gain information about the practice of infection control in oral hygiene practice. METHODS: A questionnaire based on OSHA and CDC guidelines was submitted to oral hygienists who attended a refresher course followed by a second posted questionnaire seeking information about hand hygiene practice. The questions were directed to observance of personal protection by oral hygienists and the application of procedures required for infection control in the surgery. RESULTS: 87.5% wore face masks but the proper use of facial protection, overcoats and disposable overcoats was reported by fewer than 50%. A total of 97% wore gloves and 88% changed gloves between patients. Skin reactions to gloves were reported by 26%. Only 7% had not been vaccinated against hepatitis B. The correct procedures for sterilising equipment and instruments were carried out by fewer than 50%, and 57% of participants wanted an improvement of infection control arrangements in their workplace. CONCLUSION: It is concluded that a need exists for proper practice of infection control by oral hygienists which can be remedied by interceptive and preventive education and peer pressure. PMID- 11887443 TI - HIV/AIDS: the knowledge, attitudes and behaviour of dentists in Nairobi, Kenya. AB - The purpose of this study was to assess dentists' knowledge of HIV/AIDS, as it affects them in their workplace, attitudes pertaining to universal precautions and treatment of patients with HIV/AIDS and their behaviour toward their patients. A descriptive cross-sectional survey was carried out. A 34-item questionnaire was delivered to a random sample of 145 dentists based in Nairobi, Kenya. The response rate was 72% (N = 105). Just over half the sample (53%) knew that the first AIDS patient in Kenya was reported in 1984. Ninety eight per cent knew that the main mode of transmission of HIV/AIDS in Kenya is heterosexual contact. All the respondents reported the use of gloves during clinical procedures and use of an autoclave for sterilisation of instruments was reported by more than 85%. Most dentists indicated a willingness to treat HIV/AIDS patients while those with dissenting views preferred that they be treated in dedicated clinics or academic teaching hospitals. Nearly half felt that the risk of HIV transmission in the clinic is high. The incongruity between perceived knowledge, reported practise and attitudes suggests that there is a need for continuing education courses to enable dentists to practice their profession with due care as regards patients with HIV/AIDS. In addition, courses on working with patients with HIV/AIDS should be offered so as to remove ignorance and fear. Results from this survey show that there is a fair level of knowledge as far as HIV/AIDS is concerned. The results also indicate that a greater compliance with universally accepted guidelines for infection control is needed, as it remains low for dentists working in the capital city of a country that records a rising number of new HIV/AIDS cases every day. PMID- 11887444 TI - Predictors and prognostic value of oral hairy leukoplakia and oral candidiasis in South African HIV-infected patients. AB - BACKGROUND: Oral hairy leukoplakia and oral candidiasis diseases (OHL/OC) are common clinical manifestations of HIV/AIDS. Sparse literature exists from resource-limited countries on their incidence and impact on HIV-infected patients. OBJECTIVE: To determine the predictors and prognosis of OHL/OC in HIV infected patients. METHODS: Patients were drawn from a cohort established in 1992 and prospectively followed until 1997 in the adult HIV clinics, University of Cape Town. Cox hazards regression models were fitted to determine the predictors of OHL/OC, and the association between OHL/OC and progression to AIDS and death. RESULTS: 218 patients presenting with OHL/OC at their initial clinic visit were excluded. 205/772 patients developed OHL/OC (27.8 cases/100 years). White ethnicity (hazard ratio [HR] = 1.73, 95% CI 1.23-2.33), CD4+ count < 200 cells/(L (HR = 2.55, 95% CI 1.89-3.45), total lymphocyte count < 1250 cells/(L (HR = 1.72, 95% CI 1.28-2.31) and WHO stage 3 or 4 (HR = 2.61, 95% CI = 1.93-3.53) where variables predictive of increased hazard to developing OHL/OC. OHL/OC were independently associated with hazard of AIDS (HR = 3.65, 95% CI 1.89-6.69) and death (HR = 2.12, 95% CI 1.47-4.34). CONCLUSIONS: The presence of OHL/OC in HIV infected patients provides important prognostic information, and can be used as a cost-effective tool for screening patients in therapeutic interventions in resource-limited settings. PMID- 11887445 TI - The role of primary school teachers in HIV prevention in South Africa. AB - South Africa has the fastest growing HIV epidemic in the world. The need for an intensive campaign against its spread cannot be overemphasised. Such efforts may be particularly effective if introduced prior to the onset of risk behaviour. The purpose of this study was to investigate the knowledge of grade 3 and 4 schoolteachers on HIV/AIDS and their opinion on educating their pupils about HIV prevention. A self-administered questionnaire with knowledge, perception and sociodemographic variables was sent to all 120 grade 3 and 4 teachers in the Southern Bushveld district of Northern Province. Descriptive statistics and multiple regression analysis were used to analyse the data. The response rate was 67% (N = 81) and 87.7% were females. The mean age of the respondents was 37.7 (+/ 8.7 SD) years, 55% had a 3-year teacher's training qualification and 27% had a 4 year training qualification. The average teaching experience was 12 years. Most respondents (93.8%) had knowledge of what HIV/AIDS is, but only 85.2% indicated it could be prevented. 14.8% either did not know HIV/AIDS could be prevented or were not sure. Some teachers had an incomplete understanding of the transmission and prevention of HIV/AIDS. Furthermore only 9% mentioned education as a way to prevent HIV/AIDS and 16% abstinence. Of the respondents, 58% indicated teaching HIV prevention to their pupils. Of those who do not teach HIV prevention, 41.2% believed that the pupils were too young, and 20.6% claimed non-availability of guidelines and resources as reasons for not teaching. A significant negative correlation was found between level of qualification and teaching of HIV to pupils (p < 0.05). In conclusion, many primary school teachers were found to be wanting in their HIV/AIDS knowledge. This suggests that the schoolteachers would need to be adequately trained prior to their involvement in HIV/AIDS education to pupils. Grade 3 and 4 teachers may be considered suitable to provide HIV education to their pupils, but there is a need for structured educational programmes they can follow. This study also suggests that teachers were not aware of the Department of Education's HIV policy of providing age-appropriate education to all pupils. PMID- 11887447 TI - CD4 cell monitoring for HIV/AIDS: old options; new insights. AB - Cost effective solutions are needed for laboratory monitoring that do not compromise on quality but address costs. Current recommended methods of CD4+ T cell enumeration are complex and costly. Monitoring typically utilises viral load assessment (PCR based) and CD4+ T cell counting to assess disease progression and response to therapy in HIV/AIDS. This paper reviews CD4 testing with the focus on different methods of CD4+ T cell enumeration including state-of-the-art flow cytometric testing, the advantages and disadvantages of these systems and quality control. Lastly, recent new work undertaken at the University of the Witwatersrand is discussed that addresses the problems of cost and precision and accuracy of CD4 T cell testing. PMID- 11887446 TI - Identification of Candida dubliniensis in a HIV-positive South African population. AB - Candida dubliniensis was identified as a distinctly separate species of the genus Candida in 1995. Since then the yeast has attracted considerable interest due to its prevalence in HIV/AIDS patients and its ability to develop fluconazole resistance in HIV-seropositive individuals. Although C. dubliniensis has been identified in many centres around the world it has not yet been isolated in Africa. The purpose of this study was to identify C. dubliniensis in an HIV positive population in the Western Cape, South Africa. A cohort of 50 tuberculosis patients co-infected with HIV was selected on admission to the Brooklyn Chest Hospital, Western Cape. The inclusion criteria for patients accepted for the study were: confirmed HIV seroconversion with a diagnosis of tuberculosis obtained from chest X-rays and sputum microscopy. C. dubliniensis was identified in 6 of the 50 patients accepted onto the study. The prevalence of C. dubliniensis in our study population was lower than that reported in similar North American and European studies. These results confirm the presence of C. dubliniensis in the South African HIV/AIDS population and indicate the urgent need for further investigations into the prevalence and pathogenesis of this clinically important species in both adult and paediatric HIV-positive patients. PMID- 11887448 TI - Hepatitis C infection: an overview and implications for the oral health worker. PMID- 11887449 TI - Vaccination of the HIV-1 infected child. PMID- 11887450 TI - HIV-1 drug resistance and mother-to-child transmission. AB - The use of antiretroviral drug therapies in HIV-1 infected pregnant women and their infants has resulted in significant reductions in the rates of mother-to child transmission of HIV-1. A number of drugs that target the reverse transcriptase enzyme have been tested either alone or in combination in short course regimens tailored for use in developing countries. The drug of choice is nevirapine, which is cheap, easy to administer and highly effective even following a single dose to mother and child. However, this regimen is associated with the selection of mutations associated with drug resistance. While these mutations do not compromise the ability of nevirapine to prevent mother-to-child transmission there is some concern that they may compromise future treatment options. Here we review the current data on HIV-1 drug resistance mutations and what they might mean in terms of efficacy of antiretroviral therapies to prevent mother-to-child transmission. PMID- 11887451 TI - Pneumocystis carinii pneumonia (PCP) in HIV-infected African children. PMID- 11887452 TI - The relationship between infection, inflammation, and cardiovascular disease: an overview. AB - Atherosclerotic plaques were likened histologically to healing inflammatory lesions by Russell Ross, who proposed a "response to injury" hypothesis for their formation. More recently, intraplaque inflammation has been postulated to play a role in thinning of the fibrous cap, plaque rupture, and superadded thrombosis. Potential causes for vascular injury include mechanical stress, smoke exposure, hypercholesterolemia, hyperhomocysteinemia, and chronic infection (direct, or indirect). Blood levels of inflammatory markers (e.g., C-reactive protein [CRP]; serum amyloid A [SAA]; fibrinogen; plasma viscosity; erythrocyte sedimentation rate [ESR]; leukocyte count, low serum albumin) have been associated with vascular risk factors and with prevalent and incident atherothrombotic cardiovascular disease (CVD) (coronary heart disease, [CHD]; stroke; and peripheral arterial disease). More recently, cytokines (e.g., interleukin-6 [IL 6]) and soluble adhesion molecules (e.g., intercellular adhesion molecule-1, vascular cell adhesion molecule-1) have been associated with both risk factors and disease; and offer potential therapeutic targets for nonspecific "anti inflammatory" treatment of arterial disease. Infections associated with arterial disease include specific infections (Chlamydia pneumoniae, Helicobacter pylori) and nonspecific infections (periodontal infections, respiratory tract infections). Recent meta-analyses have shown that associations of serum markers of C. pneumoniae and H. pylori with arterial disease, risk factors, or potential intermediary mechanisms for disease are weaker than was first suggested by early reports. Likewise, further studies and meta-analyses are required to evaluate the epidemiologic relationships of CVD to periodontal infection and disease and to chronic pulmonary infections and disease. The weaker the associations between chronic infections and CVD, the larger is the size of randomized controlled trials required to establish (or exclude) a preventive effect of infection treatment. While control of chronic infection in the mouth, stomach or lungs is appropriate for its local effects, proving its efficacy in prevention of CVD presents a continuing challenge to medical science. PMID- 11887453 TI - Receptor for advanced glycation end products, inflammation, and accelerated periodontal disease in diabetes: mechanisms and insights into therapeutic modalities. AB - In hyperglycemic states found in diabetics, a nonenzymatic glycation and oxidation of proteins and lipids occurs. As a result, advanced glycation end products (AGEs), particularly N epsilon-(carboxymethyl)lysine, accumulate in the plasma and tissues of diabetic subjects. This accumulation has been linked to the development of pathogenic complications of diabetes. Many of the effects of AGEs are receptor-dependent and involve a multi-ligand member of the immunoglobulin superfamily of cell surface molecules. The best characterized of these is the receptor for advanced glycation end products (RAGE), which is expressed by multiple cell types including endothelium and mononuclear phagocytes. Based on data from a variety of sources, including studies of RAGE-deficient mice, it appears that RAGE plays a central role in oral infection, exaggerated inflammatory host responses, and destruction of alveolar bone in diabetes. It is possible that antagonists of RAGE might have a valuable adjunctive therapeutic role for the management of periodontal disease found in diabetics. PMID- 11887454 TI - Insulin resistance and periodontal disease: an epidemiologic overview of research needs and future directions. AB - Poor periodontal health is known to be associated with Type 2 diabetes mellitus (DM). This relationship and underlying mechanisms are discussed elsewhere in this issue. Less is known concerning the link between the metabolic precursors to DM, including insulin resistance (IR), and its possible association with periodontitis. Indeed, there has been relatively little research to date in human populations concerning periodontal disease, IR, and the subsequent risk of chronic diseases, including DM. This paper will present an epidemiologist's view of how IR may link periodontal disease with DM and suggest several avenues of investigation to help clarify some of the outstanding issues. PMID- 11887455 TI - Periodontitis and diabetes interrelationships: role of inflammation. AB - Diabetes mellitus is a systemic disease with several major complications affecting both the quality and length of life. One of these complications is periodontal disease (periodontitis). Periodontitis is much more than a localized oral infection. Recent data indicate that periodontitis may cause changes in systemic physiology. The interrelationships between periodontitis and diabetes provide an example of systemic disease predisposing to oral infection, and once that infection is established, the oral infection exacerbates systemic disease. In this case, it may also be possible for the oral infection to predispose to systemic disease. In order to understand the cellular/molecular mechanisms responsible for such a cyclical association, one must identify common physiological changes associated with diabetes and periodontitis that produce a synergy when the conditions coexist. A potential mechanistic link involves the broad axis of inflammation, specifically immune cell phenotype, serum lipid levels, and tissue homeostasis. Diabetes-induced changes in immune cell function produce an inflammatory immune cell phenotype (upregulation of proinflammatory cytokines from monocytes/polymorphonuclear leukocytes and downregulation of growth factors from macrophages). This predisposes to chronic inflammation, progressive tissue breakdown, and diminished tissue repair capacity. Periodontal tissues frequently manifest these changes because they are constantly wounded by substances emanating from bacterial biofilms. Diabetic patients are prone to elevated low density lipoprotein cholesterol and triglycerides (LDL/TRG) even when blood glucose levels are well controlled. This is significant, as recent studies demonstrate that hyperlipidemia may be one of the factors associated with diabetes-induced immune cell alterations. Recent human studies have established a relationship between high serum lipid levels and periodontitis. Some evidence now suggests that periodontitis itself may lead to elevated LDL/TRG. Periodontitis induced bacteremia/endotoxemia has been shown to cause elevations of serum proinflammatory cytokines such as interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha), which have been demonstrated to produce alterations in lipid metabolism leading to hyperlipidemia. Within this context, periodontitis may contribute to elevated proinflammatory cytokines/serum lipids and potentially to systemic disease arising from chronic hyperlipidemia and/or increased inflammatory mediators. These cytokines can produce an insulin resistance syndrome similar to that observed in diabetes and initiate destruction of pancreatic beta cells leading to development of diabetes. Thus, there is potential for periodontitis to exacerbate diabetes-induced hyperlipidemia, immune cell alterations, and diminished tissue repair capacity. It may also be possible for chronic periodontitis to induce diabetes. PMID- 11887457 TI - Periodontal disease and diabetes mellitus: discussion, conclusions, and recommendations. PMID- 11887456 TI - Treatment of periodontal disease and control of diabetes: an assessment of the evidence and need for future research. AB - Evidence points to an increased cytokine response in type 2 diabetes, especially the proinflammatory cytokines interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha. Genetics, age, and, nutrition are important signals for this increased response and as reported more recently, infections and inflammation. Persistent elevation of IL-1 beta, IL-6, and TNF-alpha in the diabetic state have an effect on the liver, stimulate the release of acute-phase proteins, produce the characteristic dysregulation of lipid metabolism associated with type 2 diabetes, and have effects on pancreatic beta cells as well. In addition, TNF alpha, a potent inhibitor of the tyrosine kinase activity of the insulin receptor, has been implicated as an etiologic factor for insulin resistance. Collectively, the evidence supports a role for cytokine elevation in the pathophysiology and metabolic abnormalities associated with diabetes. Periodontitis is an infection that is twice as prevalent in diabetic individuals compared to non-diabetics. Porphyromonas gingivalis, one of the microorganisms responsible for this infection, is able to invade endothelial cells and is a potent signal for monocyte and macrophage activation. Thus, once established in the diabetic host, this chronic infection complicates diabetes control and increases the occurrence and severity of microvascular and macrovascular complications. Unlike treatment of acute infections, modalities of treatment for chronic infections are a matter of debate. Evidence indicates that mechanical removal of subgingival infection does not result in complete elimination of periodontal infection and consequently there is no effect on diabetes control measured as reduction in glycated hemoglobin. On the other hand, studies incorporating systemic antibiotics as adjuncts to mechanical debridement result in a reduction of P. gingivalis to nondetectable levels and a concomitant reduction in glycated hemoglobin, independent of the hypoglycemic effects of diabetes drugs or insulin. The evidence supports the notion that treatment of chronic periodontal infection is essential in the diabetic patient. Assessment of infection status in diabetic patients is fundamental for appropriate treatment decisions. PMID- 11887458 TI - The relationship between infections and adverse pregnancy outcomes: an overview. AB - Preterm birth with its subsequent morbidity and mortality is the leading perinatal problem in the United States. Infants born before the thirty-seventh week of gestation account for approximately 6% to 9% of all births, but 70% of all perinatal deaths and half of all long-term neurologic morbidity. Current approaches focus on symptomatic treatment. Despite widespread use of drugs to arrest preterm labor (tocolytics), there has been no decrease in low birth weight or preterm infants in the last 20 years. It is likely that therapy directed at preventing or treating underlying causes would be more successful. Evidence from many sources links preterm birth to symptomatic infections, for example, of the urinary or respiratory tracts. In the last decade, great interest has been generated to support the hypothesis that subclinical infection is an important cause of preterm labor. Evidence to support this may be categorized as follows: histological chorioamnionitis is increased in preterm births; clinical infection is increased after preterm birth; there is significant association of some lower genital tract organisms and infections with preterm birth or preterm premature rupture of the membranes; there are positive cultures of amniotic fluid or membranes from some patients with preterm labor and preterm birth; there are markers of infections in preterm birth; bacteria or their products induce preterm birth in animal models; and some antibiotic trials have shown a lower rate of preterm birth or have deferred preterm birth. In the last 5 years, additional exciting information has suggested that not only is subclinical infection responsible for preterm birth but also many serious neonatal sequelae including periventricular leukomalacia, cerebral palsy, respiratory distress, and even bronchopulmonary dysplasia and necrotizing enterocolitis. In sum, a large body of clinical and laboratory information suggests that subclinical infection is a major cause of preterm birth, especially those occurring before 30 weeks. This concept holds promise that new approaches can be developed to prevent prematurity. PMID- 11887459 TI - Coagulation and thrombosis in cardiovascular disease: plausible contributions of infectious agents. AB - An occlusive thrombus in the coronary arteries is the critical pathological event that immediately precedes most cases of myocardial infarction. Often the thrombus originates with a bleed from a fissured atheroma. Atheroma formation, therefore, creates risk of thrombosis; asymptomatic episodes of thrombosis and healing contribute to the pathogenesis of atherosclerosis and the development of atherosclerotic plaques. Based largely on in vitro and animal model evidence, infectious agents and their products can activate the coagulation cascade enzymatically or by up-regulating tissue factor. By initiating a procoagulant response, infectious agents can indirectly trigger a prothrombotic response. Alternatively, some microbes can directly trigger platelet aggregation in vitro and in animal models, suggesting direct prothrombotic potential in human cardiovascular disease. Activation of coagulation and thrombosis characterizes the pathological response to infectious agents in human disseminated intravascular coagulation and infective endocarditis. Given the underlying biological plausibility, the cumulative lifetime burden of chronic pathogens may be expected to create risk of atherosclerosis and thrombosis, and, indirectly, signs of cardiovascular disease. PMID- 11887460 TI - Maternal periodontitis and prematurity. Part I: Obstetric outcome of prematurity and growth restriction. AB - Oral Conditions and Pregnancy (OCAP) is a 5-year prospective study of pregnant women designed to determine whether maternal periodontal disease contributes to the risk for prematurity and growth restriction in the presence of traditional obstetric risk factors. Full-mouth periodontal examinations were conducted at enrollment (prior to 26 weeks gestational age) and again within 48 hours postpartum to assess changes in periodontal status during pregnancy. Maternal periodontal disease status at antepartum, using a 3-level disease classification (health, mild, moderate-severe) as well as incident periodontal disease progression during pregnancy were used as measures of exposures for examining associations with the pregnancy outcomes of preterm birth by gestational age (GA) and birth weight (BW) adjusting for race, age, food stamp eligibility, marital status, previous preterm births, first birth, chorioamnionitis, bacterial vaginosis, and smoking. Interim data from the first 814 deliveries demonstrate that maternal periodontal disease at antepartum and incidence/progression of periodontal disease are significantly associated with a higher prevalence rate of preterm births, BW < 2,500 g, and smaller birth weight for gestational age. For example, among periodontally healthy mothers the unadjusted prevalence of births of GA < 28 weeks was 1.1%. This was higher among mothers with mild periodontal disease (3.5%) and highest among mothers with moderate-severe periodontal disease (11.1%). The adjusted prevalence rates among GA outcomes were significantly different for mothers with mild periodontal disease (n = 566) and moderate-severe disease (n = 45) by pair-wise comparisons to the periodontally healthy reference group (n = 201) at P = 0.017 and P < 0.0001, respectively. A similar pattern was seen for increased prevalence of low birth weight deliveries among mothers with antepartum periodontal disease. For example, there were no births of BW < 1000 g among periodontally healthy mothers, but the adjusted rate was 6.1% and 11.4% for mild and moderate-severe periodontal disease (P = 0.0006 and P < 0.0001), respectively. Periodontal disease incidence/progression during pregnancy was associated with significantly smaller births for gestational age adjusting for race, parity, and baby gender. In summary, the present study, although preliminary in nature, provides evidence that maternal periodontal disease and incident progression are significant contributors to obstetric risk for preterm delivery, low birth weight and low weight for gestational age. These studies underscore the need for further consideration of periodontal disease as a potentially new and modifiable risk for preterm birth and growth restriction. PMID- 11887461 TI - Maternal periodontitis and prematurity. Part II: Maternal infection and fetal exposure. AB - Clinical data from the first 812 deliveries from a cohort study of pregnant mothers entitled Oral Conditions and Pregnancy (OCAP) demonstrate that both antepartum maternal periodontal disease and incidence/progression of periodontal disease are associated with preterm birth and growth restriction after adjusting for traditional obstetric risk factors. In the current study we present measures of maternal periodontal infection using whole chromosomal DNA probes to identify 15 periodontal organisms within maternal periodontal plaque sampled at delivery. In addition, maternal postpartum IgG antibody and fetal exposure, as indexed by fetal cord blood IgM level to these 15 maternal oral pathogens, was measured by whole bacterial immunoblots. The potential role of maternal infection with specific organisms within 2 bacterial complexes most often associated with periodontitis, conventionally termed "Orange" (Campylobacter rectus, Fusobacterium nucleatum, Peptostreptococcus micros, Prevotella nigrescens, and Prevotella intermedia) and "Red" (Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola) complexes, respectively, to prematurity was investigated by relating the presence of oral infection, maternal IgG, and fetal cord IgM, comparing full-term to preterm (gestational age < 37 weeks). The prevalence of 8 periodontal pathogens was similar among term and preterm mothers at postpartum. There was a 2.9-fold higher prevalence of IgM seropositivity for one or more organisms of the Orange or Red complex among preterm babies, as compared to term babies (19.9% versus 6.9%, respectively, P = 0.0015, chi square). Specifically, the prevalence of positive fetal IgM to C. rectus was significantly higher for preterm as compared to full-term neonates (20.0% versus 6.3%, P = 0.0002, as well as P. intermedia (8.8% versus 1.1%, P = 0.0003). A lack of maternal IgG antibody to organisms of the Red complex was associated with an increased rate of prematurity with an odds ratio (OR) = 2.2; confidence interval (CI) 1.48 to 3.79), consistent with the concept that maternal antibody protects the fetus from exposure and resultant prematurity. The highest rate of prematurity (66.7%) was observed among those mothers without a protective Red complex IgG response coupled with a fetal IgM response to Orange complex microbes (combined OR 10.3; P < 0.0001). These data support the concept that maternal periodontal infection in the absence of a protective maternal antibody response is associated with systemic dissemination of oral organisms that translocate to the fetus resulting in prematurity. The high prevalence of elevated fetal IgM to C. rectus among premature infants raises the possibility that this specific maternal oral pathogen may serve as a primary fetal infectious agent eliciting prematurity. PMID- 11887463 TI - Periodontal disease and pregnancy: discussion, conclusions, and recommendations. PMID- 11887462 TI - Current evidence regarding periodontal disease as a risk factor in preterm birth. AB - Preterm birth, resulting in babies born too little and too soon, is a major cause of morbidity. Evidence indicates that infections can be major risk factors in preterm birth. Case-control studies point to an association between periodontal infection and increased rates of preterm birth. This paper summarizes evidence to date and the strategies that ongoing intervention studies are using to answer the fundamental clinical question: can periodontal therapy reduce the risk of preterm birth? PMID- 11887464 TI - The relationship between skeletal and oral bone mineral density: an overview. AB - Is oral osteopenia (bone loss of the jaws) a component of systemic osteopenia/osteoporosis (systemic bone loss, with or without fracture) or only an accompanying manifestation of periodontal disease? Put other ways: 1) is systemic osteopenia a risk factor for periodontitis; 2) is systemic osteopenia a risk factor for oral osteopenia independent of periodontal disease; or 3) is periodontal disease the primary (exclusive) risk factor for oral osteopenia? Despite 2 decades of scientific inquiry into these questions, the answers remain elusive. PMID- 11887465 TI - Periodontal diseases and osteoporosis: association and mechanisms. AB - There is increasing evidence that osteoporosis, and the underlying loss of bone mass characteristic of this disease, is associated with periodontal disease and tooth loss. Periodontitis has long been defined as an infection-mediated destruction of the alveolar bone and soft tissue attachment to the tooth, responsible for most tooth loss in adult populations. Current evidence including several prospective studies supports an association of osteoporosis with the onset and progression of periodontal disease in humans. The majority of studies have shown low bone mass to be independently associated with loss of alveolar crestal height and tooth loss. However studies that focus on the relation of clinical attachment loss and osteoporosis are less consistent. To date, the majority of studies on the relationship between periodontal disease and osteoporosis have been hindered by small sample sizes, limited control of other potential confounding factors, varying definitions of both periodontal disease and osteoporosis, and few prospective studies where the temporality of the association can be established. Potential mechanisms by which host factors may influence onset and progression of periodontal disease directly or indirectly include underlying low bone density in the oral cavity, bone loss as an inflammatory response to infection, genetic susceptibility, and shared exposure to risk factors. Systemic loss of bone density in osteoporosis, including that of the oral cavity, may provide a host system that is increasingly susceptible to infectious destruction of periodontal tissue. Studies have provided evidence that hormones, heredity, and other host factors influence periodontal disease incidence and severity. Both periodontal disease and osteoporosis are serious public-health concerns in the United States. Prevalence of both osteoporosis and tooth loss increase with advancing age in both women and men. Understanding the association between these common diseases and the mechanisms underlying those associations will aid health professionals to provide improved means to prevent, diagnose, and treat these very common diseases. This paper reviews the current evidence on the association between periodontal disease and osteoporosis. PMID- 11887466 TI - Role of nuclear factor-kappa B (NF-kappa B) in inflammation, periodontitis, and atherogenesis. AB - Atherosclerosis, the major cause of death and disability in the United States, is a chronic disease with inflammatory components. The first objective of this review is to explain how activation of NF-kappa B contributes to atherosclerosis. The second objective is to describe a potential link between inflammation, activation of NF-kappa B, and periodontitis. The nuclear transcription factor NF kappa B controls the expression of many genes linked to atherogenesis including those involved with inflammation. We hypothesize that one unifying mechanism in this complex disease is the activation of NF-kappa B. The mechanism(s) that activates NF-kappa B in atherogenesis is unknown and the effect of inhibiting NF kappa B activation on atherogenesis is untested. Periodontal disease has now been established as a risk factor for atherosclerosis and its thrombotic complications. It is unknown if periodontal disease contributes to the initiation or progression of atherosclerosis. We hypothesize that the chronic and intense inflammatory response accompanying periodontal disease produces an excess burden of circulating mediators of inflammation that initiate or exacerbate the inflammatory components of atherogenesis. Further understanding of the mechanisms involved in the activation of NF-kappa B in atherosclerosis could lead to important therapeutic applications especially as it relates to the impact of periodontitis. PMID- 11887467 TI - The periodontal-systemic connection: implications for treatment of patients with osteoporosis and periodontal disease. AB - Osteoporosis and osteopenia may influence periodontal disease and tooth loss. Medications such as hormone replacement therapy and nutritional supplements that are used to prevent or treat osteoporosis have been evaluated for beneficial effects on oral health in a small number of human studies. Hormone replacement therapy (HRT), which slows the rate of bone loss at skeletal sites such as the hip and spine, also appears to reduce the rate of alveolar bone loss in postmenopausal women. HRT use is consistently associated with greater tooth retention and a reduced likelihood of edentulism in studies of elderly women. The number of studies on the effects of calcium or vitamin D intake on oral outcomes is limited, but suggest that higher intake levels are associated with reduced prevalence of clinical attachment loss and lower risk of tooth loss. Data from a prospective study of oral health in men show a similar association between higher calcium intake and reduced alveolar bone loss. The number of teeth with progression of alveolar bone loss over a 7-year period was significantly lower among men whose calcium intake was at least 1,000 mg per day, compared to men with a calcium intake below this level. Future studies should confirm these findings and evaluate the oral effects of new medications for osteoporosis. If confirmed, the implications for dental professionals may include an expanded array of medications for the treatment of periodontal disease and a greater emphasis on nutrition education for patients. PMID- 11887468 TI - Osteoporosis and periodontitis: discussion, conclusions, and recommendations. PMID- 11887469 TI - The role of inflammatory and immunological mediators in periodontitis and cardiovascular disease. AB - Epidemiological studies have implicated periodontitis (PD) as a risk factor for development of cardiovascular disease (CVD). Persistent infections such as periodontitis induce inflammatory and immune responses which may contribute to coronary atherogenesis, and, in conjunction with other risk factors, may lead to coronary heart disease (CHD). In this review, mechanisms are described that may help explain the association between periodontal infections and CHD. Periodontal diseases are bacterial infections associated with bacteremia, inflammation, and a strong immune response, all of which may represent significant risk factors for the development of atherogenesis, CHD, and myocardial infarction (MI). Several mechanisms may participate in this association, including those induced by oral organisms, and those associated with host response factors. This review will focus on host factors. Oral pathogens and inflammatory mediators (such as interleukin [IL]-1 and tumor necrosis factor [TNF]-alpha) from periodontal lesions intermittently reach the bloodstream inducing systemic inflammatory reactants such as acute-phase proteins, and immune effectors including systemic antibodies to periodontal bacteria. This review will describe the potential role of various inflammatory as well as immunologic factors that may play a role in periodontitis as a possible risk factor for CHD. PMID- 11887470 TI - Role for periodontal bacteria in cardiovascular diseases. AB - BACKGROUND: Several epidemiological studies as well as a recent animal model approach have suggested a role for periodontal diseases in the development of cardiovascular disease (CVD). This relationship could be mediated by inflammatory responses induced by periodontal pathogens as well as direct interaction of these organisms with cardiac tissue. METHODS: In order to explore these possibilities, the effects of the periodontal pathogen Porphyromonas gingivalis on cellular events proposed to play a role in CVD were investigated. RESULTS: P. gingivalis, as well as its outer membrane vesicles (OMV), was able to induce foam cell formation (an important characteristic of CVD) in the murine macrophage cell line J774 A.1. This property appears to be mediated by the lipopolysaccharide (LPS) fraction of the cells. Several other oral bacteria were also able to induce foam cell formation. Furthermore, since the rupture of the fibrous cap of plaque appears to be an important factor in acute coronary syndrome, it was demonstrated that P. gingivalis 381 degraded fibrous caps isolated from autopsy samples. In addition, it was observed that strain 381 strongly induced matrix metalloproteinase (MMP)-9 protease activity, implicated in plaque rupture, from the J774 A.1 macrophages. Finally, strain 381 was able to enhance monocyte chemoattractant protein-1 (MCP-1) and NADH oxidase expression from endothelial cells. CONCLUSIONS: Therefore, P. gingivalis exhibits several properties which could play a role in CVD as mediators of LDL oxidation, foam cell formation, and rupture of atherosclerotic plaque. PMID- 11887471 TI - Candidate genes as potential links between periodontal and cardiovascular diseases. AB - Recent epidemiological associations between periodontal disease and cardiovascular disease have led to a search for biological mechanisms that explain the associations. Genetic factors that influence biological processes involved in both diseases represent one of the potential mechanisms that may link periodontitis and cardiovascular disease. At present, several candidate genes have been investigated in one of the diseases but not the other. Although there are limited data to support a specific candidate gene as the explanation for observed associations between the 2 diseases, a few candidates look promising. One candidate that influences inflammation, interleukin-1 gene polymorphisms, has been associated with periodontal disease and cardiovascular disease. This review will consider biological mechanisms and genes that may be reasonable candidates for an etiological mechanism that influences the clinical characteristics of both periodontal disease and cardiovascular disease. PMID- 11887472 TI - The relationship between infections and chronic respiratory diseases: an overview. AB - This paper's goal is to review the relationship between infections and chronic respiratory disease, with particular reference to periodontal disease. The link between oral diseases in general, periodontal disease, and respiratory disease remains somewhat controversial. However, with cooperation between dentistry and medicine, the nature of the connection between dental and medical pathology can be better defined. An overview of respiratory disease and some of the factors that can contribute to respiratory infection is presented below, with special reference to infections related to aspiration. PMID- 11887473 TI - Epidemiologic associations between periodontal disease and chronic obstructive pulmonary disease. AB - The nature of the relationship of periodontal disease to a number of systemic health outcomes, including chronic obstructive pulmonary disease (COPD), remains unclear. Various causal mechanisms have been proposed to explain the observed epidemiologic associations between periodontal diseases and respiratory diseases. We have reviewed the epidemiologic and clinical evidence for this association. The methodologic approach we have taken is based on a structured systematic review of the indexed biomedical literature on these subjects. The primary focus of this review was on the analysis of periodontal health status measures and their association with COPD, which includes chronic bronchitis and emphysema. We found that a paucity of published results exist on this specific relationship and those which do exist typically represent secondary analyses of existing data sets. Nevertheless, the epidemiologic evidence identified in this systematic review indicates that worse periodontal health status is associated with an increased risk of COPD, with odds ratios ranging from 1.45 to 4.50 (significant at the 95% confidence interval). However, it is possible that residual confounding by tobacco smoking may account in part for the observations. A causal association between periodontal health status and risk of COPD, although biologically plausible, remains speculative. Randomized controlled trials will be required in order to address the question of causality and to better understand the biological basis of these epidemiologic associations. PMID- 11887475 TI - Periodontitis and respiratory diseases: discussion, conclusions, and recommendations. PMID- 11887474 TI - Oral bacteria and respiratory infection: effects on respiratory pathogen adhesion and epithelial cell proinflammatory cytokine production. AB - Several microbiologic and epidemiologic studies have suggested an association between dental plaque, poor oral health, and respiratory diseases such as nosocomial pneumonia and chronic obstructive pulmonary disease (COPD). A number of hypotheses are suggested to help explain how oral bacteria may participate in the pathogenesis of respiratory infection. Resident bacteria in oral secretions are likely aspirated along with respiratory pathogens and may affect the adhesion of the later organisms to the respiratory epithelium. Preliminary studies performed in our laboratory suggest that oral bacteria may modulate the adhesion of respiratory pathogens to epithelial cell lines. In addition, oral bacterial products or cytokines in oral/pharyngeal aspirates may stimulate cytokine production from respiratory epithelial cells, resulting in recruitment of inflammatory cells. The resulting inflamed epithelium may be more susceptible to respiratory infection. Further preliminary data are presented that some species of oral bacteria may induce the release of proinflammatory cytokines from epithelial cell lines to an extent similar to that seen for respiratory pathogens. PMID- 11887476 TI - The association between periodontal diseases and cardiovascular diseases: a state of-the-science review. AB - Early case-control and cross-sectional studies demonstrating associations between chronic periodontitis and cardiovascular disease (CVD) were quickly followed by secondary analyses of data available from existing longitudinal studies, which indicated that individuals with periodontitis, as determined by clinical measures, were at greater risk for CVD events. Many of these studies contained large numbers of subjects and were adjusted for traditional risk factors. Within the last 18 months, one case-control study and one longitudinal study have reported finding positive associations that were not statistically significant. The earlier studies stimulated a number of studies focused on identifying potential biological mechanisms that might underlie this association. While still early in that process, such studies have implicated a systemic role for oral microorganisms and for the quality and quantity of the host inflammatory response as key biologic processes that may underlie the association of CVD with the clinical manifestation of periodontitis. It is a positive development when changes in our knowledge regarding biologic mechanisms result in reevaluation of past studies, and this reevaluation leads to new studies that incorporate the design elements demanded by this new knowledge. In that spirit, we conclude that all longitudinal studies reported to date can be characterized as follows: none were initially designed to actually test the association of interest; almost all were restricted to clinical measures of periodontitis to index the exposure and lacked measures of infectious burden and host response; and they used a variety of cardiovascular clinical events to index the outcome and did not include subclinical measures of atherosclerosis. In addition, the longitudinal studies that failed to show a significant association between periodontitis and CVD used the least sensitive and crudest clinical measures of periodontal disease. Based upon the current state-of-the-science, all previous studies should be viewed as lacking sufficiently sensitive and comprehensive measures of periodontal disease as a systemic exposure. Since the potential health care impact of this relationship might be extensive, it is time to enter the next phase of research by conducting molecular epidemiology studies that are appropriately designed to test our current understanding of the molecular and cellular mechanisms involved. PMID- 11887477 TI - The relationship between periodontal diseases and diabetes: an overview. AB - Diabetes mellitus, caused by the malfunction of insulin-dependent glucose and lipid metabolism, presents with the classical triad of symptoms: polydypsia, polyuria, and polyphagia which are often accompanied by chronic fatigue and loss of weight. Complications of diabetes mellitus include retinopathy, nephropathy, neuropathy, and cardiovascular disease. Periodontal diseases are infections affecting the periodontium and resulting in the loss of tooth support. The association between diabetes mellitus and periodontitis has long been discussed with conflicting conclusions. Both of these diseases have a relatively high incidence in the general population (diabetes 1% to 6% and periodontitis 14%) as well as a number of common pathways in their pathogenesis (both diseases are polygenic disorders with some degree of immunoregulatory dysfunction). On the one hand, numerous reports indicate a higher incidence of periodontitis in diabetics compared to healthy controls, while other reports fail to show such a relationship. Clarification of this dilemma is occurring as the diagnostic criteria for periodontitis and diabetes mellitus improve, controlled studies with increased sample sizes are carried out, and the studies take into account major confounding variables that impact on the pathogenesis of both diseases. Current studies tend to support a higher incidence and severity of periodontitis in patients with diabetes mellitus. The overview looks at the bidirectional relationship between periodontitis and diabetes. An analysis of the National Health and Nutrition Examination Survey (NHANES) III data set confirms the previously reported significantly higher prevalence of periodontitis in diabetics than in non-diabetics (17.3% versus 9%). The analysis of the data also shows that the prevalence of diabetes in patients with periodontitis is double that seen in the non-periodontitis patients (12.5% versus 6.3%) and that this difference is also statistically significant. The pathogenesis of the 2 diseases is reviewed with an emphasis on common genetic and immune mechanisms. On the basis of the overview, 2 hypotheses for testing the relationship between periodontitis and diabetes are discussed. The first proposes a direct causal or modifying relationship in which the hyperglycemia and hyperlipidemia of diabetes result in metabolic alterations that may then exacerbate bacteria-induced inflammatory periodontitis. The second hypothesis proposes that a fortuitous combination of genes (gene sets) could result in a host who, under the influence of a variety of environmental stressors, could develop either periodontitis or diabetes or both. PMID- 11887479 TI - Iatrogenic illness: a primer for nurses. AB - Iatrogenic illness is a term that is used frequently but not clearly understood. This overview of diseases incurred as a consequence of medical treatment explores evolving definitions and the epidemiology of these problems. In addition, a number of strategies to reduce the incidence of these illnesses and the implications of this problem for nurses are presented. PMID- 11887478 TI - Bidirectional interrelationships between diabetes and periodontal diseases: an epidemiologic perspective. AB - This review evaluates evidence for a bidirectional relationship between diabetes and periodontal diseases. A comprehensive Medline search of the post-1960 English language literature was employed to identify primary research reports of relationships between diabetes and periodontal diseases. Reports included in the review on the adverse effects of diabetes on periodontal health (DM-->PD) were restricted to those comparing periodontal health in subjects with and without diabetes. Review of adverse affects of periodontal infection on glycemic control included reports of periodontal treatment studies and follow-up observational studies in which changes in glycemic control could be assessed. Observational studies reporting DM-->PD provided consistent evidence of greater prevalence, severity, extent, or progression of at least one manifestation of periodontal diseases in the large majority of reports (supportive evidence in 44/48 total reviewed; 37/41 cross-sectional and 7/7 cohort). Additionally, there were no studies reviewed with superior design features to refute this association. Treatment studies provided direct evidence to support periodontal infection having an adverse, yet modifiable, effect on glycemic control. However, not all investigations reported an improvement in glycemic control after periodontal treatment. Additional evidence to support the effect of severe periodontitis on increased risk for poorer glycemic control comes from 2 follow-up observational studies. The evidence reviewed supports viewing the relationship between diabetes and periodontal diseases as bidirectional. Further rigorous, systematic study is warranted to establish that treating periodontal infections can be influential in contributing to glycemic control management and possibly to the reduction of the burden of complications of diabetes mellitus. PMID- 11887481 TI - Patient education handout. Key elements of photosensitity. PMID- 11887480 TI - Evaluation and management of the patient with photosensitivity. AB - Evaluation of patients with photosensitivity includes a detailed history, physical examination, phototests, photopatch tests, and other laboratory tests as appropriate. The epidemiology, clinical features, diagnosis, and management of the more common idiopathic photodermatoses, namely, polymorphous light eruption, chronic actinic dermatitis, and solar urticaria will be reviewed. A brief overview of phototoxicity, photoallergy, and photoprotection is discussed with further elaboration upon the principles of phototherapy and its utility in treating idiopathic photodermatoses. PMID- 11887482 TI - What's your assessment? Granuloma fissuratum. PMID- 11887483 TI - Incidence of CTCL in Vietnam veterans. AB - The causative factors of cutaneous T-cell lymphoma (CTCL) are unclear. Exposure to herbicides has been linked to the development of other lymphomas. Three Vietnam Veterans with CTCL treated at a photopheresis unit in New Jersey report positive histories of exposure to Agent Orange, a herbicide used during the war. PMID- 11887484 TI - Caring for patients with hearing impairments. AB - Ensuring that handicapped persons are not denied goods and services, including health care, because of their disability is the essence of The Americans With Disabilities Act. Nurses can help assure that patients with hearing impairments receive proper, quality care in their dermatology practices. PMID- 11887485 TI - Cutaneous T-cell lymphoma. AB - The "Clinical Snapshot" series provides a concise examination of a clinical presentation including history, treatment, patient education, and nursing measures. Using the format here, you are invited to submit your "Clinical Snapshot" to Dermatology Nursing. PMID- 11887486 TI - Obstructive nephropathy: pathophysiology, diagnosis, and collaborative management. PMID- 11887487 TI - The LifeSite hemodialysis access system: implications for the nephrology nurse. AB - A new subcutaneous device--the LifeSite Hemodialysis Access System (Vasca, Inc., Tewksbury, MA)--was designed to overcome limitations of transcutaneous dialysis catheters and is now available for use in the United States. A fully implantable device, the LifeSite System provides immediate, reliable, high-flow vascular access. The durable stainless steel and titanium LifeSite valve is implanted in a subcutaneous tissue pocket, typically below the clavicle. It is connected to a biocompatible silicone cannula that is tunneled to a central vein. The device is cannulated using a virtually pain-free, buttonhole technique. The valve is designed to allow cleansing of the valve, valve pocket, and buttonhole site with 70% isopropyl alcohol before and after each use to minimize the risk of infection. One hundred fifty-day results from an ongoing multicenter trial designed to compare the LifeSite System to a Tesio Cath dialysis catheter are reviewed. These results demonstrate that the LifeSite System is associated with statistically significant higher blood flow rates (p < 0.001) and lower rates of adverse events (p < 0.0004), infection (p < 0.032), and thrombolytic infusions (p < 0.044) than a standard dialysis catheter. The positive clinical experience with the LifeSite System carries significant implications for the dialysis team, indicating that this subcutaneous, pain-free route to vascular access may offer a safer, more effective bridge to a permanent arteriovenous (AV) access than a tunneled dialysis catheter. Potential implications of these benefits include improved outcomes, greater convenience for patients, improved efficiency and time management for the nursing staff, along with reduced direct and indirect costs related to vascular access management. PMID- 11887488 TI - Health promoting behaviors, quality of life, and hospital resource utilization of patients receiving kidney transplants. AB - Despite the value of kidney transplantation for patients with renal failure, transplantation is sometimes accompanied by untoward consequences and considerable resource utilization. The identification of modifiable factors contributing to resource utilization is, therefore, important. This exploratory study of kidney transplant patients had two purposes: (a) to describe health promoting behaviors and health-related quality of life (HRQOL) pretransplant and hospital resource utilization posttransplant and (b) to investigate relationships between these and other variables. Subjects were 44 patients who (before transplantation) completed the Health-Promoting Lifestyle Profile (HPLP) to quantify health-promoting behavior and the Medical Outcome Study Short Form (SF) 36 to measure HRQOL. Hospital resource utilization was characterized using transplant length of stay and charges and posttransplant emergency department visits, readmissions, and charges. The HPLP scores of the subjects were similar to those reported for middle-aged adults and patients with other diseases. The SF 36 subscale and summary scores were lower than those reported for the general population. Hospital resource utilization varied widely. Significant correlations were found between 52.9% of the HPLP and SF-36 scores. However, no HPLP score correlated significantly with any utilization measure. Of the SF-36 measures, only vitality correlated significantly with most utilization measures (greater vitality was associated with less posttransplant utilization). While many aspects of health-promoting behavior and HRQOL are related before transplant, neither HPLP, SF-36, nor any other measured variable strongly and consistently predicted hospital resource utilization. PMID- 11887489 TI - Hyporesponse to Epoetin alfa: patients at risk. Case study of the anemic patient. AB - A wide spectrum of clinical data has consistently demonstrated improvements in survival, hospitalization, and other factors affecting quality of life in dialysis patients who maintain hemoglobin levels between 11 and 12 g/dL, and most U.S. dialysis facilities generally target this range for their patients. Recent data indicate that several factors, including time on dialysis, Epoetin alfa dosing practices, peritoneal dialysis modality, younger age, African American race, and some comorbidities, may increase the risk for lower hemoglobin levels. Although additional clinical studies are required to fully define the factors that may contribute to lower hemoglobin levels in these patients, nephrology nurses should be aware of the potential need to modify therapeutic approaches in these subpopulations to ensure that all patients have the opportunity to attain and maintain target hemoglobin levels. PMID- 11887490 TI - Aranesp (darbepoetin alfa): a new erythropoiesis-stimulating protein. PMID- 11887491 TI - Hypertension in the hemodialysis patient: nursing considerations. PMID- 11887492 TI - Positive changes on the horizon for Nephrology Nursing Journal. PMID- 11887493 TI - Arterial endothelin-1 in interstitial lung disease patients with pulmonary hypertension. AB - The aim of this study was to evaluate the plasma arterial levels of Endothelin-1 (ET-1) and their relationship with hypoxia and pulmonary hypertension (PH) in patients with interstitial lung disease (ILD). Incremental cycle ergometry was performed in all patients up to maximal capacity. ET-1 levels during exercise (2.2 +/- 0.36 pgr/ml) were significantly higher than at rest (1.73 +/- 0.37 pgr/ml) (p < 0.001). ET-1 levels were also significantly correlated with arterial oxygen (PaO2) (r = -0.935, p < 0.001) and mean pulmonary arterial pressure (Ppa) (r = 0.657, p < 0.001). Increased pulmonary and peripheral blood levels of ET-1 have been described in, and postulated to contribute to, the pathophysiology of several lung diseases. In agreement with this, in the current study, the plasma arterial levels of ET-1 were also found to be significantly elevated in patients with various interstitial lung disorders during exercise, especially in those with severe hypoxia and pulmonary hypertension. PMID- 11887494 TI - How can we best read exhaled nitric oxide flow curves in asthmatic children? AB - Orally exhaled nitric oxide (NO) levels are increased in children with asthma and thought to reflect the local inflammatory events in the airways. NO production in the lower respiratory airway is reflected in the plateau values of the NO curve, recorded while the patient is performing a slow vital capacity manoeuvre. In young patients, however, plateau values may be difficult to obtain, because the slow vital capacity manoeuvre is often terminated prematurely. In the present study, 60 steroid-naive atopic asthmatic children and 17 normal age-matched controls were asked to perform a slow vital capacity manoeuvre, during which fractional exhaled NO (FEno) levels were measured and evaluated as: a) FEno plateau levels of last part of exhalation (NO plateau); b) FEno peak values, c) area under the FEno curve (AUC). Thirteen out of the 60 steroidnaive patients were reevaluated after a short course of inhaled corticosteroid treatment. Independently of the type of data analysis, FEno values of asthmatics were significantly higher than those observed in normal controls (P < 0.001, each comparison). In addition, possibly because of upper airway NO contamination, FEno peak values were significantly higher than FEno plateau levels in asthmatic patients and in control subjects (P < 0.001, each comparison). Both in asthmatics and controls, highly positive correlations were observed between: a) FEno plateau and peak values (r > 0.7, P < 0.01, each correlation), b) FEno plateau and AUC values (r > 0.7, P < 0.01, each correlation) and c) FEno peak and AUC values (r > 0.9, P < 0.001, each correlation). In asthmatic patients, the three types of data analysis were equally sensitive in detecting the decrease in FEno levels induced by inhaled steroid therapy (P < 0.05, each comparison), with a good correlation between the three data analyses (r > 0.5, P < 0.05, each correlation). Thus, although quantitatively different, comparable data reflecting airway inflammation can be obtained evaluating FEno plateau, FEno peak, and area under the curve, on account of possible upper airway contamination in FEno peak, FEno plateau should be preferred to measure lower airway NO production. PMID- 11887495 TI - The causes of massive hemoptysis. AB - In a group of 656 patients examined for hemoptysis of unknown etiology the average amount of expectorated blood in individual diagnostic groups was evaluated and the patients with massive hemoptysis (> 100 ml/24 hrs) were selected. The hemoptysis was evaluated as massive in 53 patients (8%), being most frequent in patients with chronic anatomical changes of lung parenchyma (22/64) and active tuberculosis (4/26). The fact that massive hemoptysis endangers above all patients with bronchiectasis and other anatomical disorders of the lung parenchyma was evident through the comparison of the average maximal amount of expectorated blood, which was significantly higher in this group (78 ml/24 hrs) than in the majority of other diagnoses (total average 22 ml/24 hrs). In conclusion, the risk of sudden massive hemoptysis should be kept in mind in these patients even if only minimal bleeding occurs and energetic cautionary measures should be implemented right from the onset of bloody expectoration. PMID- 11887496 TI - Management of malignant pleural effusions. AB - Lung and breast cancer are responsible for the majority of malignant pleural effusions. The diagnosis of a malignant pleural effusion signifies a limited survival for most patients. During their final months, dyspnea is the most common symptom and requires palliation. A decision relating to palliation and the modality of therapy should be based on total assessment of the patient and not a single variable. Local treatment remains the most common and effective palliation. Assessing the response to therapeutic thoracentesis determines the degree of relief of dyspnea and the time-course of recurrence. Lack of a beneficial effect suggests the patient may have a trapped lung, atelectasis, lymphangitic carcinomatosis, or tumor embolism. Short-term chest tube drainage has variable results and is not recommended. Chemical pleurodesis through a standard chest tube or small-bore catheter is a commonly used and effective treatment. Talc slurry consistently produces the highest success rates, followed by the tetracyclines and bleomycin. Although acute respiratory failure has been reported following talc pleurodesis, these episodes represent a very small percentage of the total reported cases of talc poudrage and slurry pleurodesis. Whether acute respiratory failure is directly related to talc in the absence of other risk factors remains unclear. Other possible causes for acute respiratory failure following pleurodesis include re-expansion pulmonary edema, excessive premedication, severe comorbid disease, and sepsis from unsterile talc or poor chest tube technique. Factors that need to be considered before recommending chemical pleurodesis include response to therapeutic thoracentesis, general health of the patient, performance status, pleural space elastance, the primary malignancy, and pleural fluid pH. Chronic indwelling catheters have been shown to be effective alternatives to chemical pleurodesis. Pleuroperitoneal shunting can provide palliation to patients with a trapped lung, a malignant chylothorax, or others who have failed pleurodesis. Parietal pleurectomy should be reserved only for patients who have failed chemical pleurodesis or have a trapped lung with an expected survival > 6 months. To provide the highest quality of life for patients with malignant pleural effusions, the least invasive, morbid and costly therapy should be used. Success of the initial procedure is important, as repeat procedures are associated with additional hospitalization, patient discomfort, and increased expense; therefore, the selection of patients for palliation and the modality utilized is critical to avoiding further hardship to the patient. PMID- 11887497 TI - Daytime somnolence. Basic concepts, assessment tools and clinical applications. AB - Excessive somnolence is a common symptom, with a prevalence of 10 to 20% in a general population. However, physicians seldom ask their patients about sleep complaints. The internal biological clock drives the balance between sleepiness and alertness, generating circadian rhythms, with "physiological" increases of somnolence, especially at mid-day and before the habitual bed time. Excessive somnolence is a subjective feeling of an imperious need of sleep in unusual time and environmental conditions. Sleep deprivation, sleep fragmentation and to a lesser degree hypoxia are believed to be the main mechanisms leading to excessive somnolence. Excessive somnolence increases the risk of car accidents, deteriorates health status and quality of life and might increase mortality. Excessive somnolence is associated with many diseases such as obstructive sleep apnoea syndrome. Excessive sleepiness can be assessed by visual scales or questionnaires, the best known being the Epworth sleepiness scale. Objective tests in somnolent patients assess the sleep-wake balance disturbances. The most widely used tests are the multiple sleep latency test (MSLT), the maintenance of wakefulness test (MWT) and the Oxford sleep resistance (OSLER) test. These tests measure the time to sleep emergence in different conditions. PMID- 11887498 TI - Prophylaxis of chronic lung disease after premature birth. AB - Chronic lung disease (CLD) following premature birth is a common adverse outcome of neonatal intensive care. It particularly occurs in infants born at very early gestations who are exposed to the dual insults of volutrauma and oxygen toxicity. Many possible prophylactic strategies have been investigated in randomized trials, but none have been demonstrated to be both effective and safe. High frequency oscillation, if a high volume strategy is pursued, reduces the incidence of CLD, but it remains unclear whether this is at the expense of an increase in intracranial pathology particularly in very immature infants. Corticosteroids administered systemically in the first two weeks after birth reduce CLD and may favourably impact on mortality; however, the long term consequences on pulmonary function and neurodevelopmental outcome of this therapy are of concern. PMID- 11887499 TI - Skeletal muscle dysfunction in chronic obstructive pulmonary disease. AB - During the last decade evidence has been accumulated on the role of skeletal muscle dysfunction in reducing exercise capacity and affecting the quality of life of patients with chronic obstructive pulmonary disease (COPD). An appreciable body of research has helped to identify morphological and biochemical alterations, physiological consequences, and possible therapeutic interventions. There are, however, still many areas of uncertainty. For example it is not clear how much of the alterations are within the muscle itself or the consequence of the altered environment in which the muscle works. Similarly it is not clear how much of the impairment is simply due to aging and chronic inactivity. Another key issue is the possible additive effect of drugs often used in COPD patients, such as steroids, beta 2-agonist and cyclosporin. A specific additional layer of complexity comes from nutritional considerations and in particular loss of muscle mass which not infrequently accompanies severe disease and even greater exercise intolerance. Studies on the effects of training or other therapeutic interventions have shown that muscle dysfunction is partially reversible. There is, however, a clear need for studies based on cellular and molecular methods aimed to clarify the role of factors such as oxidative stress, inflammation and nutritional deficiencies on skeletal muscle structure and function. The focus of this review is to highlight the current knowledge on skeletal muscle dysfunction in COPD and briefly summarize the possible therapeutic implications. PMID- 11887500 TI - Severe and near-fatal asthma in children and adolescents. AB - Near-fatal asthma attacks occur rather frequently in asthmatic patients during childhood and adolescence. Major risk factors appear to be the severity and/or treatment difficulties, which may be related to the characteristics of the disease but also to mistaken or delayed diagnosis, poor compliance to treatment, and psychosocial problems. Indeed, difficult-to-control asthma symptoms can be caused by other disorders mimicking asthma, by unidentified exacerbating factors, by low compliance with the prescribed therapy, by inability to use the devices correctly (metered-dose inhalers, dry powder inhalers, nebulizers) and by low perceptiveness of airway obstruction. Children and adolescents with severe, difficult-to-treat asthma should receive specialist assessment and follow-up. Pharmacologic intervention should be combined with a detailed education and action plan, designed to get the disease under control but also to meet patient' and parents desires. PMID- 11887501 TI - Exacerbations of asthma: addressing the triggers and treatments. AB - Airway inflammation is known to play a crucial role in chronic asthma and has led to the effective use of anti-inflammatory treatments to control chronic asthma. Despite these improvements exacerbations of asthma continue to occur in subjects with stable disease compliant with treatment. Viral respiratory tract infections have been found to be associated with the majority of asthma exacerbations in both children and adults. Evidence is emerging that viral infections may alter the inflammatory infiltrate present in chronic asthma with a more heterogenous neutrophil/eosinophil infiltrate. The implications are that the pathogenesis of acute asthma may differ from that present in chronic asthma in important ways that may influence the effectiveness of treatment. Inhaled corticosteroids are effective in controlling chronic airway inflammation in asthma but appear not to be as effective in acute asthma. This may mean that new directions will be needed to target specifically the airway inflammation in virus-induced acute asthma. PMID- 11887502 TI - Transbronchial needle aspiration of central and peripheral nodules. AB - Wang and Terry first described the technique of sampling of mediastinal lymph nodes using a flexible bronchoscope (FB) in 1983. Since then, the scope of transbronchial needle aspiration (TBNA) has increased enormously, and its value for diagnosis of central and peripheral lung lesions, even in the absence of endobronchial disease, is now recognized. Improvements in the diagnostic yield of TBNA aspirates, and increasing knowledge of predictors of a positive aspirate, has reduced the need for mediastinoscopy, and occasionally thoracotomy, with benefits in terms of reduced healthcare costs and improved patient welfare. Despite the fact that TBNA is a minimally invasive procedure, also for the staging of lung cancer, it unfortunately remains underutilized. This review aims to give the reader an overview of the indications, outcome data, technical considerations, and advantages of routinely performing TBNA. The technical suggestions to improve the yield from TBNA will hopefully provide greater understanding of this modality and help the pulmonologist to incorporate it in daily practice with more confidence. PMID- 11887503 TI - Which biopsies in diffuse infiltrative lung diseases and when are these necessary? AB - Different biopsy techniques are used to obtain appropriate tissue for diagnostic pulmonary pathology. Traditionally open lung biopsies were more often used in the USA, whereas transbronchial biopsies prevailed in many European countries. However, the type of biopsy especially in interstitial lung diseases (ILD) should be selected not on traditional grounds, but on a scientific rationale. In progressive ILD where the pathologic diagnosis is based on the recognition of different patterns at different process stages, an open or video-assisted thoracoscopic lung biopsy is preferred, whereas in most granulomatous pneumonias transbronchial or even bronchial biopsies may suffice. For many other ILDs no recommendations exist, but will be tentatively given. In addition comprehensive clinical information should be provided for the pathologist, because this is the prerequisite for a qualitative diagnosis, which means not only a descriptive answer, but also an etiologically based one. PMID- 11887504 TI - The role of information, education and treatment in tobacco control. AB - Because tobacco use is both an individual and a social behaviour, the efficacy of information, education and treatment addressed to smokers is greatly influenced by the inhibiting or facilitating effects of the environment and the social values given to smoking and to not smoking. The environment is a function of the evolution of tobacco use and the extent of tobacco control efforts. Information is the fuel for legislative and regulatory measures. These and public health education campaigns set the stage for building motivation and confidence to stop smoking and reinforce the social value of non-smoking. As individuals desire, and become prepared for, change, treatments help them address their needs. The role of information, education and treatment are interconnected. Each plays a different and necessary part in building and reinforcing social change and individual behaviour change. They are all needed to move populations and the individuals in those populations as rapidly as possible away from smoking. PMID- 11887505 TI - The economic and social costs of lung cancer and the economics of smoking prevention. AB - Lung cancer is a major cause of the premature loss of life and the costs to society of lung cancer are significant. Since smoking is the major cause of lung cancer, the main prevention strategy is to reduce smoking. Interventions to reduce smoking have been shown to be cost effective, from population measures such as tax and advertising restrictions through to individual or group treatments. Different groups of the population may well benefit from different interventions and there could be synergistic effects from a comprehensive programme. Treatment for smokers is an important part of any programme and especially important to prevent longer term smokers contracting lung cancer. However, most health care systems have not been successful in ensuring that smoking cessation interventions have a high priority. It should be an urgent task to ensure that the small additional resources for this work are fully reimbursed and incentives introduced to reap both major health gains and future reductions in health costs. PMID- 11887506 TI - WHO evidence based recommendations on the treatment of tobacco dependence. PMID- 11887507 TI - Smoking cessation: the role of bupropion among new pharmacologic agents. AB - Cigarette smoking is the leading cause of preventable death and the major cause of respiratory diseases. According to the World Health Organization, it is also a disease in itself, which can be treated with a behavioural approach. Nicotine replacement therapy (NRT) has been established during the last decades as a safe and effective pharmacologic treatment for this disease. Now, new pharmacologic treatments are being tested. This paper reviews the most promising ones among these new drugs: nicotine agonists and antagonists, anxiolitics, antidepressants. Among them, bupropion seems the most promising and is analysed on the basis of the existing scientific literature. According to the literature, bupropion is an effective and safe way to obtain smoking cessation, independently of its antidepressant action. However, it can have a few important adverse effects and has some contraindications. Caution should therefore be used in prescribing it to some types of patient, but the drug is active even at half the recommended dosing. In the future other drugs, active on the central nervous system, will be studied for smoking cessation, and the option of using bupropion and NRT together should be further explored. PMID- 11887508 TI - A maxillary fibro-osseous lesion: differential diagnosis and case report. PMID- 11887509 TI - Ethical dilemma. PMID- 11887510 TI - Oral and maxillofacial pathology case of the month. Lipoma. PMID- 11887511 TI - Development of a quantitative method to monitor the effect of a tooth whitening agent. AB - This study demonstrated a quantitative method for assessing the effect of a tooth whitening agent. Forty human teeth were stained with a tea solution, and randomly assigned to two groups (A, B) of twenty teeth. The teeth were subsequently treated with either sodium hypochlorite (NaOCL) or deionized distilled water (DDW) by intermittent immersion (60 seconds on each occasion) in a 1:10 dilution of NaOCL (group A) or DDW (group B). Prior to whitening and following each immersion, the color of the teeth at the stained spot was measured using ShadeEye Ex Dental Chroma Meter and quantitative light-induced fluorescence (QLF). ShadeEye-Ex instantly gave a numerical value for the stain intensity, chroma (C), which is the average of three measurements taken automatically by the machine. QLF gave a quantitative value for the stain, delta Q (% mm2), following analysis of the fluorescence image of the tooth. Immersion was stopped after four readings when one specimen, in group A, was observed to have regained its natural color. There was a good correlation between C and delta Q with either NaOCL (Pearson correlation coefficient (r) = 0.974; p < 0.05) or DDW (r = 0.978; p < 0.05). With NaOCL, an inverse relationship observed between stain measurements, C (Linear fit correlation (R) = -0.982; p < 0.05) or delta Q (R = -0.988; p < 0.05) and exposure time correlated to a linear fit, but not with DDW. ANOVA showed a significant difference between the means (n = 20) of the reading at the measurement intervals (0, 60, 120 and 180 seconds) for both C (p < 0.001) and delta Q (p < 0.001) with NaOCL but not with DDW. In conclusion, the study highlighted the potential of ShadeEye-Ex Dental Chroma Meter as a tool for the quantitative assessment of the gradual change in shade of discolored teeth by tooth whitening products. PMID- 11887512 TI - Effect of demineralization and remineralization on microhardness of irradiated dentin. AB - The aim of this in vitro study was to evaluate the effects of irradiation and fluoridation on the demineralization and remineralization patterns of root dentin. From the cervical regions of 84 bovine incisors, each of four dentinal blocks were prepared and randomly assigned to four groups: 1) no irradiation; 2) irradiation of specimens up to 60 Gy (2 Gy/d, 5 d/w); 3) no irradiation, but fluoridation of specimens for 5 min/d with Elmex Gelee; and 4) irradiation and daily fluoridation of specimens for 5 min/d. Subsequently, the specimens were demineralized for 2, 4, 6 or 8 days with acidulated hydroxyethylcellulose (n = 21), and Knoop hardness numbers (KHN) were determined before, as well as after the demineralization period. Then seven specimens from each group were fluoridated with one of three fluoride gels (Elmex Gelee [1.25%], STOP [0.4%], Fluoridgel [1.25%]; 2 x 15 min/d, 10 d), and immersed in synthetic saliva at a temperature of 37 degrees C. Finally, KHN for all specimens were determined. Irradiation resulted in a significant decrease in microhardness. There was a reduction in microhardness with increasing demineralization time in all groups. The highest percentage decrease in microhardness could be observed with group 1. Due to the preceding decrease of KHN in the irradiated specimens (group 2), these samples showed less percentage reduction in microhardness during demineralization when compared to group 1 (ANOVA; p < 0.001). The decrease in microhardness was significantly hampered by fluoridation in the non-irradiated, as well as in the irradiated samples (groups 3 and 4; p < 0.001). The remineralization with Fluoridgel resulted in the greatest increase in microhardness. It is concluded that demineralization can be hampered by regular fluoride application in irradiated dentin. However, due to the considerable irradiation effect, this benefit might be negligible. PMID- 11887514 TI - Dental considerations in sucralose use. AB - Sucralose is a new type of non-caloric, high-intensity sweetener recently approved for use by the U.S. FDA. Its availability may expand the number of palatable, low-sugar foods and beverages currently on the market. A series of studies has been conducted to assess whether sucralose has cariogenic potential. These include an examination of oral bacterial metabolism, experimental caries in animal models, and the effect of sucralose-containing solutions on human plaque pH in situ. The sum of these studies demonstrates that sucralose is non cariogenic. Sucralose-based sweeteners that contain bulking ingredients, which allow them to pour and measure more like sugar, do have cariogenic potential due to the presence of added fermentable carbohydrate; however, the data suggest that both the currently marketed sucralose granular and packet products are less cariogenic than sugar. Thus, when used to replace sugar, both sucralose and the tested sucralose-based sweeteners may be useful in the dietary management of caries. PMID- 11887513 TI - Microleakage of a packable composite associated with different materials. AB - The aim of this in vitro study was to evaluate the extent of marginal leakage with class II composite resin restorations associated with different materials. Twenty extracted sound molars were disinfected and stored in 0.9% saline solution. Four standardized class II cavities were prepared in each tooth, with the gingival margin 1 mm below the cementum-enamel junction. The teeth were then randomly divided into five groups of 16 restorations each: 1) Prime & Bond NT + Surefil; 2) Prime & Bond NT + Dyract Flow + Surefil; 3) Prime & Bond NT + Flow-it + Surefil; 4) Prime & Bond NT + Dyract AP + Surefil; and 5) Fuji II LC + Prime & Bond NT + Surefil. After a storage time of seven days, the restorations were finished and polished. Then specimens were submitted to thermocycling (500 cycles, 5 degrees-55 degrees C, 15 s-dwell time) and immersed in 0.5% methylene blue solution for 24 hours. They were washed and vertically sliced through the middle of the restoration. Both surfaces were evaluated in the gingival margin and one score (0-3) was assigned under a stereomicroscope (40x magnification) by two examiners. The frequency of the scores was evaluated by Kruskal-Wallis and Mann-Whitney tests for pairwise comparisons. None of the associations prevented dye penetration. The statistical analysis showed similar dye penetration among groups 1, 2, 3 and 4 (p > 0.05). The results of penetration in group 5 were significantly lower compared to the other groups (p < 0.05). It was concluded that the use of Fuji II LC as a base material led to the best sealing of the gingival margins based upon the lowest degree of microleakage observed. PMID- 11887515 TI - A clinical investigation into the effect of toothbrush wear on efficacy. AB - It is generally recommended that toothbrushes should be replaced after three months' use in order to maintain efficacy. This clinical investigation employed a single-use, cross-over study and a three-month parallel-group study to investigate the effect of toothbrush wear on plaque and gingival health. Toothbrushes were artificially worn using a laboratory wear machine to simulate three months of clinical toothbrush use. Results from the single-use study showed that both the new and the worn toothbrushes significantly reduced whole mouth, marginal and approximal plaque scores from pre- to post-brushing (p < 0.0001). The new brush was, however, significantly more effective than the worn brush, demonstrating 13.4%, 11.0%, and 17.0% greater plaque reduction for whole mouth, marginal and approximal sites, respectively (p < 0.0001). Results from the three month study confirmed this finding, with significant differences in plaque reduction (p < 0.0001) between the new and worn toothbrushes at 6 and 12 weeks for all sites. A significant difference (p < 0.0001) between the groups was also found for mean whole mouth gingivitis scores; this difference favoring the new brush at both 6 and 12 weeks. Examination of hard and soft oral tissues revealed no significant difference between the new and the worn brushes with respect to safety. It is concluded that a worn toothbrush is less effective than a new toothbrush for plaque removal and control of gingivitis. PMID- 11887517 TI - Comparative efficacy of two battery-powered toothbrushes on dental plaque removal. AB - A number of clinical studies have consistently demonstrated that power toothbrushes deliver superior plaque removal compared to manual toothbrushes. Recently, a new power toothbrush (Crest SpinBrush) has been marketed with a design that fundamentally differs from other marketed power toothbrushes. Other power toothbrushes feature a small, round head designed to oscillate for enhanced cleaning between the teeth and below the gumline. The new power toothbrush incorporates a similar round oscillating head in conjunction with fixed bristles, which allows the user to brush with optimal manual brushing technique. The objective of this randomized, examiner-blind, parallel design study was to compare the plaque removal efficacy of a positive control power toothbrush (Colgate Actibrush) to an experimental toothbrush (Crest SpinBrush) following a single use among 59 subjects. Baseline plaque scores were 1.64 and 1.40 for the experimental toothbrush and control toothbrush treatment groups, respectively. With regard to all surfaces examined, the experimental toothbrush delivered an adjusted (via analysis of covariance) mean difference between baseline and post brushing plaque scores of 0.47, while the control toothbrush delivered an adjusted mean difference of 0.33. On average, the difference between toothbrushes was statistically significant (p = 0.013). Because the covariate slope for the experimental group was statistically significantly greater (p = 0.001) than the slope for the control group, a separate slope model was used. Further analysis demonstrated that the experimental group had statistically significantly greater plaque removal than the control group for baseline plaque scores above 1.43. With respect to buccal surfaces, using a separate slope analysis of covariance, the experimental toothbrush delivered an adjusted mean difference between baseline and post-brushing plaque scores of 0.61, while the control toothbrush delivered an adjusted mean difference of 0.39. This difference between toothbrushes was also statistically significant (p = 0.002). On average, the results on lingual surfaces demonstrated similar directional scores favoring the experimental toothbrush; however these results did not achieve statistical significance. In conclusion, the experimental Crest SpinBrush, with its novel fixed and oscillating bristle design, was found to be more effective than the positive control Colgate Actibrush, which is designed with a small round oscillating cluster of bristles. PMID- 11887516 TI - Efficacy and safety of BrushPicks, a new cleaning aid, compared to the use of Glide floss. AB - The objective of this double-blind, four-week clinical study was to evaluate the efficacy of BrushPicks, a new cleaning aid, and Glide floss on the reduction of plaque area, gingivitis and bleeding on probing, and to monitor safety when these products were used in addition to toothbrushing with an ADA-Accepted toothbrush (Oral-B P35) and an ADA-Accepted fluoride-containing dentifrice (Crest Regular). No special instructions on or supervision of product use was conducted, other than requesting twice-a-day (morning and evening) use of the assigned products. Following a baseline examination, 63 qualifying adult male and female subjects from the Philadelphia, Pennsylvania area were randomized into two groups. Subjects were also told to use their assigned dental aid after each toothbrushing. Examinations for efficacy and safety were repeated after two and four weeks' use of the products. Sixty-two subjects completed all aspects of the study. There were no untoward side effects attributed to product use, reported or observed, at the two- or four-week examination times. At baseline, there were no significant differences in plaque, gingivitis or bleeding on probing mean scores between the BrushPicks and Glide floss groups. At the two- and four-week evaluation times, both the BrushPicks and Glide floss had numerically lower plaque scores compared to baseline levels. The only statistically significant reduction (p < 0.01) was in the BrushPicks group, comparing the week two mean with the baseline value. Gingivitis (GI) at four weeks was statistically (p < 0.05) lower in the BrushPicks group as compared to the Glide floss mean value. When the changes in scores from baseline to two weeks and to four weeks were assessed, the mean GI score for the Glide floss group was significantly lower at two weeks (p < 0.01) compared to baseline, and also from two weeks to four weeks (p < 0.001). The change in mean GI score for the Glide floss group from baseline to four weeks was also significant statistically (p < 0.001). When the changes in mean GI scores for the BrushPicks group were assessed, there was a significant decrease from baseline to two weeks (p < 0.001), from two weeks to four weeks (p < 0.001), and from baseline to four weeks (p < 0.001). For bleeding on probing, when the baseline to two- and four-week mean values were compared, only the BrushPicks produced significant (p < 0.001) decreases. At two and four weeks, the BrushPicks group mean bleeding on probing scores were significantly (p < 0.05 0.01) lower than the Glide floss group scores. At the end of this four-week study, the BrushPicks product was significantly more effective than Glide floss in the reduction of gingivitis and bleeding on probing, important attributes of soft-tissue health. PMID- 11887518 TI - Busting UCR. PMID- 11887519 TI - Coding for routine periodontal treatment. PMID- 11887520 TI - Avoiding Medicaid fraud. PMID- 11887521 TI - Non-pharmacologic behavior management techniques used with pediatric dental patients. PMID- 11887522 TI - Considerations for monitoring pediatric sedation. PMID- 11887523 TI - Doctor or Mister: who's or whose calling? PMID- 11887524 TI - Taking advantage of the tax relief act: estate planning. PMID- 11887525 TI - It's National Children's Dental Health Month--make a difference in the life of a child. PMID- 11887526 TI - Introducing Dr. Paul Landman, CDS president for 2002. PMID- 11887527 TI - No missing pieces: midwinter meeting put together for you. PMID- 11887528 TI - Associateships: a practice arrangement that can incorporate dollars and sense! PMID- 11887529 TI - AIDS turns 21. PMID- 11887530 TI - The use of magnification in a preventive approach to caries detection. AB - OBJECTIVE: The aim of this study was to demonstrate the effect of low-powered magnification on the accuracy of caries detection and to compare it to the accuracy of unaided vision. METHOD AND MATERIALS: Five dental models were prepared with extracted, unrestored, human permanent premolars, molars, and canines. Dental examinations were undertaken in simulated clinical conditions by seven dentists using both unaided and magnified vision. A true diagnosis was obtained by histologic sectioning, thereby allowing diagnostic accuracy to be calculated. RESULTS: The sensitivity of diagnosis, representing the percentage of diseased sites found correctly, was significantly greater when magnification was used. There was no statistically significant difference in the specificities, or percentages of correctly identified healthy sites, between magnification and unaided vision. CONCLUSION: Magnification, although not perfect, improved significantly on the accuracy of diagnosis and can therefore be recommended for caries detection. PMID- 11887531 TI - Comparison of atraumatic restorative treatment and conventional cavity preparations for glass-ionomer restorations in primary molars: one-year results. AB - OBJECTIVE: The purpose of this study was to compare the success rates of glass ionomer cement restorations placed with the atraumatic restorative treatment approach and conventional cavity preparation methods. METHOD AND MATERIALS: Two encapsulated, high-strength, esthetic conventional glass-ionomer cements were placed in 82 Class I and 53 Class II atraumatic restorative treatment and conventional cavity preparations, and an encapsulated amalgam alloy was placed in 32 Class I conventional preparations, in vital primary molars of 60 Chinese children aged 7 to 9 years. RESULTS: The atraumatic restorative treatment preparations, made with hand instruments only, took approximately 50% longer to complete than did the preparations completed with conventional rotary instrumentation. After 1 year, there were no amalgam failures. For the glass ionomer cement restorations, when the atraumatic restorative treatment method was used, significantly better survival rates were found for Class I (92.9%) than for Class II (64.7%) cavity preparations. There was also a strong trend for relatively better survival rates for the conventional cavity preparation method (86.7%) than for the atraumatic restorative treatment (64.7%) method for Class II cavity preparations. However, both the atraumatic restorative treatment and conventional methods appeared equally effective for Class I preparations. CONCLUSION: In a clinic setting, the use of atraumatic restorative treatment hand instruments for cavity preparation is more time consuming, and the method may also provide less mechanical retention and/or bulk of glass-ionomer cement for some Class II preparations in primary molars than does the use of conventional rotary instruments. PMID- 11887532 TI - An in vitro study of coronal microleakage around bonded amalgam coronal-radicular cores in endodontically treated molar teeth. AB - OBJECTIVE: The aim of this study was to compare the coronal microleakage of conventional and bonded amalgam coronal-radicular (Nayyar) restorations on endodontically treated molar teeth, because coronal seal is a major factor in the long-term success of endodontic treatment. METHOD AND MATERIALS: Forty extracted human molar teeth were root-filled and prepared for coronal-radicular amalgam restorations. Four groups of 10 teeth were restored with Tytin amalgam and Vitrebond, Superbond D Liner II, Panavia 21, or no adhesive agent. The teeth were placed in India ink for 1 week, and then demineralized and rendered transparent. The ink penetration was assessed with a coded scoring system. RESULTS: The bonded amalgam groups produced significantly less leakage than did the nonbonded group. No statistically significant differences in leakage were detected among the bonded amalgam groups. CONCLUSION: To prevent the reinfection of the endodontically treated molar, it may be preferable to restore the tooth immediately after obturation by employing a bonded amalgam coronal-radicular technique. PMID- 11887534 TI - Pulp-dentin biology in restorative dentistry. Part 6: Reactions to restorative materials, tooth-restoration interfaces, and adhesive techniques. AB - A number of defense mechanisms may be activated in conjunction with restorative dental treatment. Tissue changes that lead to reduced dentinal permeability are important to minimize pulpal reactions. The healing potentials of the pulp-dentin complex are significant, and restorative dentistry is dependent on the available defense reactions. Clinicians should intentionally include considerations of these reactions in their treatment planning and during restorative treatment. PMID- 11887533 TI - Electrical and dye leakage comparison of three root-end restorative materials. AB - OBJECTIVE: The purpose of this study was to assess three modern root-end restorative materials with electrical and dye leakage tests. METHOD AND MATERIALS: Thirty-three human canine teeth were prepared and filled with gutta percha and sealer. Stainless steel rods were inserted into the root canals as anodes, and the teeth were varnished. The apical 3 mm of each tooth was resected, and 3-mm root-end preparations were made ultrasonically. Mineral trioxide aggregate, Super EBA, and IRM were used to restore 10 teeth each, and three teeth were varnished as controls. Following 24 hours' setting in blood, the specimens were placed in 1% potassium chloride electrolyte, and leakage was recorded electrically for 70 days. The teeth were then submerged in methylene blue dye for 72 hours, sectioned longitudinally, and scored for leakage by six examiners. RESULTS: In both tests, the mineral trioxide aggregate restorations leaked significantly less than the IRM and Super EBA restorations. Super EBA showed significantly less leakage than did IRM restorations in the electrochemical test but not in the dye leakage experiment. The teeth sealed with mineral trioxide aggregate performed similarly to the varnished negative control teeth. CONCLUSION: The results of this study indicated that mineral trioxide aggregate provides a superior seal in root-end-restorations. PMID- 11887535 TI - Pathologic migration--spontaneous correction following periodontal therapy: a case report. AB - Periodontal disease is often associated with pathologic migration, which becomes an esthetic concern. A 17-year-old girl developed increasing gaps among her maxillary incisors. She had gingival enlargement in the palatal maxillary anterior region. The central incisors had pathologically migrated, resulting in a 2-mm diastema. Periodontal treatment was planned and completed. Following periodontal treatment, there was "spontaneous" repositioning of the central incisors. The 6-month follow-up revealed no change or deterioration of the periodontal condition. The patient was referred for orthodontic closure of the remaining diastema between the central and lateral incisors. PMID- 11887536 TI - Dental concepts in the Unified Medical Language System. AB - OBJECTIVE: Currently, no comprehensive, controlled vocabulary for dentistry is available. The objective of this study was to determine how well the Unified Medical Language System, the largest repository of concepts and terms in biomedicine, represents dental concepts. METHOD AND MATERIALS: The dental subset of concepts was extracted from Unified Medical Language System using the software program APEX (APplication for the EXtraction of domain-specific concepts). The relationships contained in the Unified Medical Language System Metathesaurus were used to locate the concepts related to 12 seed terms. The Encyclopedie Medico Chirurgicale (513 dental terms) and the Diagnostic Codes developed by Leake et al (124 terms) were compared to the dental subset. Terms were classified as exact match, related term, or no match. The resultant matching characteristics were compared to those determined by the National Library of Medicine/Agency for Health Care Policy and Research Large Scale Vocabulary Test. RESULTS: The dental subset of Unified Medical Language System contained 948 concepts. The Encyclopedie Medico-Chirurgicale and the Diagnostic Codes exhibited similar matching characteristics for exact match (61.6% and 58.9%, respectively) and related term (38.0% and 32.2%, respectively). For no match, the matching frequencies were significantly different (P < .001). CONCLUSION: The Unified Medical Language System may be a comprehensive source of terms suitable for various representation requirements in dentistry. PMID- 11887537 TI - Cowden's syndrome: a case report. AB - Cowden's syndrome, a rare genodermatosis of autosomal-dominant inheritance with variable expressivity, is characterized by a combination of ectodermal, mesodermal, and endodermal hamartomas that may involve the skin, mucous membranes, breasts, gastrointestinal tract, and thyroid. A 26-year-old woman who presented for replacement of her teeth, all of which had been extracted because of rapidly progressive periodontitis. She was diagnosed with Cowden's syndrome based on mucocutaneous abnormalities, thyroid involvement, and abnormalities of the skeletal and genitourinary systems. The clinical significance and differential diagnosis of this disease are highlighted. PMID- 11887538 TI - Paget's disease (osteitis deformans). PMID- 11887539 TI - School-related expenses, living expenses, and income sources for graduate students in nurse anesthesia programs. AB - Nurse anesthesia programs (NAPs) are the highest priced programs for graduate students compared with 7 other nursing master's degree programs. Not only are nurse anesthesia programs expensive, but also most students are encouraged by the policies within their individual programs to terminate full-time employment before matriculation. The purpose of this study was to determine school-related and living expenses, as well as the income and sources of income for graduate students in the second year of their NAP. To obtain the information, a student cost survey was designed and administered to participants attending NAPs across the United States during the 2001 school year. In addition, total degree costs were analyzed using a cost model assessing 4 components: educational costs, living expenses, net income foregone, and loan costs. The results showed that total degree costs incurred by graduate students in NAPs to complete their nurse anesthesia education totals $173,007. The analysis of the sources of income showed the following sources were used by respondents: guaranteed student loans; a spouse's income; agreements with future employers; stipends from universities, hospitals, and/or the military; grants; family support; and self-income. Completing a nurse anesthesia education program is expensive, although the expected return on the investment is high. Nevertheless, the expense may keep qualified graduate students from entering NAPs. PMID- 11887540 TI - Diagnosis and treatment of tension pneumothorax under anesthesia: a case report. AB - An 80-year old woman with a history of tracheal stenosis, tracheostomy, and 3 months of increasing respiratory distress underwent tracheal dilatation under general anesthesia with jet ventilation. Tracheal dilatation was successfully performed via suspension laryngoscopy and jet ventilation. During emergence the patient developed decreased oxygen saturation, hypotension, and respiratory distress, requiring intubation and ventilatory support. During tracheostomy, anterior chest subcutaneous emphysema led to a diagnosis of tension pneumothorax. Chest tube placement facilitated tracheostomy and improved ventilatory and circulatory parameters. This article discusses the diagnosis and treatment of a tension pneumothorax under general anesthesia. Jet ventilation, spontaneous rupture of blebs or bullae, surgical trauma, or barotrauma are the 4 most likely explanations for a tension pneumothorax in this patient. Jet ventilation can cause pneumothorax, pneumomediastinum, or subcutaneous emphysema. Chronic obstructive pulmonary disease may cause blebs or bullae, which might rupture when exposed to positive pressure ventilation. Tissue trauma during dilatation or tracheostomy may cause a pneumothorax when positive pressure ventilation is employed. Barotrauma from high peak inspiratory pressure, rigid bronchoscopy, dilatation procedure, or jet ventilation may cause a pneumothorax. Prompt diagnosis and treatment will markedly decrease associated morbidity and mortality. PMID- 11887541 TI - The morphine-sparing effect of metoclopramide on postoperative laparoscopic tubal ligation patients. AB - Metoclopramide traditionally has been used as a prokinetic and antiemetic, but recently it also has been investigated as an agent to enhance analgesic efficacy. No definitive studies have been undertaken to determine whether metoclopramide can decrease postoperative analgesic requirements. The present study examined the effects of the administration of metoclopramide on the postoperative opioid analgesic requirements and pain intensity scores of patients following laparoscopic bilateral tubal ligation under general anesthesia. Fifty-six subjects were enrolled in this double-blind, placebo-controlled, prospective study. Subjects were randomized into 2 groups, a control placebo group (n = 30) and an experimental metoclopramide group (n = 26), and were assessed for pain intensity using a numeric rating scale. Assessments were recorded at 3 time intervals. Total postoperative opioid requirements also were recorded. Numeric rating scale pain scores were noted to be similar in both groups at all 3 time intervals examined. However, total postanesthesia care unit (PACU) morphine requirements were significantly higher in the placebo group than the metoclopramide group (P = .031). This study demonstrated that metoclopramide administered preoperatively can significantly decrease morphine requirements in the PACU but had no impact on pain intensity as rated by numeric rating scale pain scores. PMID- 11887542 TI - Prevalence of and reasons for preoperative tobacco use. AB - Smoking cigarettes has an impact on all aspects of the perioperative anesthetic. It is not known whether patients are typically educated regarding these effects. Eighty-one patients completed a questionnaire concerning smoking behavior in the 24 hours before surgery. Variables measured were smoking history, tobacco addiction, and preoperative education. Chi-square analysis was used. Of 81 participants, 66 (81%) smoked tobacco within 24 hours of surgery. Thirty-seven patients received no instructions to stop smoking, and only 2 patients abstained on their own. Of the 44 patients counseled not to smoke, 12 abstained from tobacco before operation. Thus, with counseling, the cessation rate was approximately 5 times greater (chi 2 = 7.0, P = .008). A second correlation was seen when the patients were informed about tobacco's risks related to anesthesia. The smoking rate decreased from 15% to 4%, a 4-fold decrease (chi 2 = 15.3, P = .0001). The results indicate patients who smoke are not routinely informed of the risks of tobacco use or the benefits of abstinence before surgery. Counseling has a positive impact on the patient's smoking behavior in the 24 hours preceding surgery. Anesthesia providers and surgeons have a renewed obligation to instruct patients not to smoke before surgery. PMID- 11887543 TI - Safety in the use of compressed air versus oxygen for the ophthalmic patient. AB - Oxygen, routinely administered during surgery to avoid hypoxia, poses risks including increased likelihood of surgical room fires and predisposition to retinal phototoxicity in patients. Compressed air to supplement ventilation may be safer than oxygen. The purpose of this study was to determine whether hypoxia occurs more frequently when compressed air replaces supplemental oxygen during ophthalmic surgery. A convenience sample of 111 patients was randomly assigned to receive supplemental oxygen (group 1) or compressed air (group 2). Patients with serious cardiac or pulmonary disease were excluded. Blood oxygen levels were monitored during surgery by pulse oximetry. Oxygen was administered to all group 2 patients whose oxygen saturation fell to less than 90% or by more than 5% below baseline. No differences were observed between groups in age, ASA classification, type of surgery, or anesthetic drugs or doses. Minor, but statistically higher oxygen values were observed in group 1. The frequency with which oxygen saturation decreased below 90% or below 5% of baseline was similar in both groups. Supplemental oxygen is not required routinely in selected patients undergoing ophthalmic surgery. By using compressed air, the risk of operating room fires and retinal phototoxicity may be reduced. PMID- 11887544 TI - Herbal medicines and possible anesthesia interactions. AB - Herbal medicines are biochemically active compounds that have the potential to interact with drugs used in anesthesia. Lack of herbal standardization makes definitive diagnosis of herb-anesthesia interaction difficult. However, identified herbal use and an understanding of possible interactions can alert the anesthesia provider and raise suspicion of possible herb-related complications. In a recent survey, 22% of patients undergoing surgery reported herbal medicine use. Most patients using herbal remedies fail to report this use to their healthcare providers. Herbal medicine use is not routinely addressed during the preoperative interview. The interviewer should include open-ended questions such as: "What herbal or vitamin supplements do you currently take?" The purpose of this review is to give an industry overview and look at commonly used herbal medicines, focusing on those with the greatest potential for anesthetic and operative complications. PMID- 11887545 TI - Microbial growth on the anesthesia machine. AB - The purpose of this study was to determine the amount of microbial growth that develops on the anesthesia machine after a full day's use in the operating room. This descriptive bacteriology study is relevant to anesthesia practice because of the proximity of the oropharynx and multiple body fluids to anesthesia equipment and the potential for cross-contamination to patients and staff. The Wilcoxon signed rank test was used to evaluate the change in colony-forming units (CFUs) before and after use of equipment. The resulting P value of 0.12 indicated that the observed CFU increase was not statistically significant at the .05 level. The study identified many organisms that survive on the anesthesia machine tabletop, namely, coagulase-negative Staphylococcus, Bacillus, alpha Streptococcus, Acinetobacter, Staphylococcus aureus, and gram-negative rods. Several were expected to be found; however, alpha Streptococcus, Acinetobacter, S aureus, and gram-negative rods are pathogenic organisms causing respiratory infections and bacteremia, especially in patients with compromised conditions. Terminal cleaning methods may have changed during the course of the study, thereby contributing to the volume of microbes present before use and distorting the change in the number of CFUs before and after use. PMID- 11887546 TI - The use of nalmefene for intrathecal opioid-associated nausea in postpartum patients. AB - The aim of this study was to compare the severity of nausea and incidence of emesis in laboring parturients who received intravenous nalmefene or placebo following an intrathecal opioid (ITO). We randomly assigned 60 ASA class I or II multiparous women to receive nalmefene or placebo. Subjects received fentanyl, 25 micrograms, and morphine, 250 micrograms, intrathecally on request for analgesia. Thirty minutes after vaginal delivery, the experimental group received nalmefene, 20 micrograms, and the placebo group received an equivalent volume of normal saline intravenously. Visual analog scale nausea scores and data about episodes of emesis were obtained during labor and during the first 24 hours postpartum. There were no significant differences in age, weight, duration of labor, volume of intravenous fluids infused, time from last meal to delivery, or time from administration of the ITO to injection of the study drug. There were no significant differences in mean visual analog scale nausea scores or frequency of emesis for any time interval. Nalmefene, 20 micrograms, given intravenously within 30 minutes of vaginal delivery does not significantly reduce the nausea and vomiting associated with the use of ITOs for labor analgesia. PMID- 11887547 TI - A pathway toward safer anesthesia: stereochemical advances. PMID- 11887548 TI - CRNAs as independent contractors. PMID- 11887549 TI - [Acute coronary syndrome--state of the art 2002]. PMID- 11887550 TI - [Etiology of acute coronary syndrome--unstable plaque]. AB - Our perception of the mechanisms underlying the acute complications of atherosclerosis has significantly changed in the last decade. Most coronary thromboses result from a rupture or a fissure in the protective fibrous cap of atherosclerotic plaque, but less common from a superficial erosion. The extent of thrombosis is a determinant of the clinical picture of acute coronary syndromes. Until now, we held a high grade stenosis responsible for the vast majority of acute coronary syndromes. However, current findings establish the relevance of qualitative aspects of plaques as important determinants of the vulnerability of plaques, i.e. the risk to cause acute complications. Among morphologic and functional features of plaques, inflammation has emerged as a leading pathophysiologic mechanism. In addition to local effect of inflammation at the level of the unstable plaque itself, systemic factors of the inflammatory response may alter the thrombogenicity of a vulnerable plaque. Knowledge of the role of inflammation helps us to understand the mechanisms by which therapeutic efforts can reduce clinical events. The clinical benefits of dietary modifications, pharmacotherapy with statins, and ACE inhibitors may be due in part to an anti-inflammatory action. PMID- 11887551 TI - [Acute coronary syndrome and myocardial infarct--new definitions]. AB - Until recently a general consensus existed for the clinical entity diagnosed as myocardial infarction using the world health organisation (WHO) definition. According to the WHO definition myocardial infarction was defined by a combination of two of three typical characteristics: typical symptoms, rise of cardiac enzymes (CK, CK-MB), and a typical ECG pattern involving the development of Q waves. New insights into the development of acute myocardial infarction, the superiority of the biochemical characteristics of cardiac troponin assays over CK and CK-MB measurements in blood, and new therapeutic concepts made a new definition of myocardial infarction, e.g. of the acute myocardial infarction, necessary. Timing of the diagnosis of myocardial necrosis is of outmost importance relative to the time of observation (acute, evolving, healing, healed MI), as is the classification of the extent of myocardial damage (microscopic, small, medium or large). The term "acute coronary syndrome" (ACS) has been established as a working diagnosis for choosing the appropriate therapeutic strategy. In patients with ACS and ST elevation ischemia (STEMI ACS, true posterior ischemia inclusive) as well as in patients with presumably new LBBB, immediate reperfusion therapy should be performed (primary PTCA or thrombolytic therapy), whereas in patients with ECG changes other than ST elevation or new LBBB (NSTEMIACS) additional antiplatlet therapy on top of aspirin and heparin is indicated. In contrast to the acute phase of infarction when troponin in blood often is not detectable yet, the diagnosis of definitive myocardial infarction is based primarily on troponin elevation. Hard criteria for established infarction are the development of pathologic Q waves or healing or healed myocardial necrosis in pathology; troponin may be normal then, depending of time relapsed. PMID- 11887552 TI - [Diagnosis and risk stratification of acute coronary syndrome in general practice and in the hospital]. AB - Acute coronary syndromes usually present as acute chest pain but a manifestation with atypical symptoms or entirely without symptoms makes rapid diagnosis of this potentially lethal syndrome difficult and may lead to delay of the appropriate therapy. The role of the practitioner in this situation is complex since early exclusion of an acute coronary syndrome may be very difficult with the means available in a private practice; on the other hand the early hours which are critical for early therapy may be lost in a private practice. In the early phase of an acute coronary syndrome, diagnoses and risk stratification are based primarily on history, clinical presentation, ECG and biological markers. In the hospital, the time course of these parameters, functional tests for diagnosis and risk stratification as well as determination of left ventricular function and coronary angiography have additional relevance. Recent developments are the availability of highly sensitive and specific biological markers (Troponins), the new classification in acute coronary syndrome with and without ST-elevation and the identification of inflammatory processes in the coronary artery, which have added importantly to our understanding of the acute coronary syndrome. The availability of very early therapy and the possibilities of telemedicine have the potential to influence the pre-hospital diagnostic strategy and management of acute coronary syndromes. PMID- 11887553 TI - [Management of acute myocardial infarct with ST segment elevation]. AB - In the management of acute myocardial infarction with ST segment elevation, the primary goal is rapid reperfusion of the initially occluded infarct-related coronary artery. This may be achieved either by catheter using direct coronary angioplasty or by medical therapy in form of thrombolysis. These two methods, their concomitant treatment strategies as well as most recent results of combination therapy are presented and discussed. This results in recommendations for the management of these patients which have to take into account logistic possibilities and minimize time delays. PMID- 11887554 TI - [Medicamentous management of acute coronary syndrome without ST segment elevation]. AB - Recent advances in the recognition and the treatment of acute coronary syndromes (ACS) have lead to an improvement in patient survival and definition of newer guidelines. Current strategies for the treatment of patients with non-ST elevation ACS include anti-ischemic and antiplatelet medications. While aspirin, beta-blockers, heparin and nitrates are still common practice, the advent of newer anticoagulants (low molecular weight heparins) and antiplatelet agents (glycoprotein llb/IIIa inhibitors and thienopyridines like ticlopidin and clopidogrel) and, possibly, aggressive lipid lowering with statins have added significant benefits to the treatment options with a better prognosis for these patients. Moreover, aggressive medical strategies seem to be justified not only in high-risk patients but also in those that undergo an early invasive approach. PMID- 11887555 TI - [Which patients with acute coronary syndrome without ST segment elevation should be handled invasively/interventionally when?]. AB - Until recently the therapeutic approach in patients with acute coronary syndrome without ST-segment elevation focused on medical stabilization. Usually cardiac catheterization and revascularisation were performed later only if the stabilized patient had provoked ischemia. Since angioplasty became safer with the introduction of coronary stents, and since new potent antithrombotic agents (i.e. tienopyridines or glycoprotein-receptorIIb/IIIa-antagonists) have been developed, early angioplasty has become much more effective. Moreover major progress has been made in risk-stratification, which allow an individual therapeutic strategy for each patient according to his risk-status. We discuss the most important randomised clinical trials comparing a conservative versus an invasive strategy and introduce a new algorithm for risk-stratification und therapy in acute coronary syndromes without ST-elevation. PMID- 11887556 TI - [Secondary prevention after acute coronary syndrome--role of modern drug therapy]. AB - Treatment of acute coronary syndrome is under rapid progress. Nevertheless, the early complication rate remains high. Standard antithrombotic treatment is Aspirin 100 mg/d. Patients with elevated risk should be treated with Aspirin and Clopidogrel if primary invasive strategy ist not intended. Independent of the cholesterol level, statins should be given in the early phase of acute coronary syndrome. Dose adaptation is recommended after three months corresponding to the national guidelines. Mainly in diabetes mellitus, additional treatment with an ACE inhibitor lowers the overall cardiovascular risk also in patients without arterial hypertension or congestive heart failure. Six months after the acute event, risk stratification should be adapted. PMID- 11887557 TI - [Case of the month. Problems in diagnosis of acute coronary syndrome]. AB - Three cases of patients with suspected acute coronary syndrome are presented. The importance of the clinical evaluation is stressed in relation to ECG changes and determinations of new necrosis markers. PMID- 11887558 TI - [What is the role of sunscreens in the prevention of melanoma?]. PMID- 11887559 TI - [Summary of the Skin Cancer Prevention Campaign 2000 of the Suisse league against cancer "Protect me! Your skin"]. PMID- 11887560 TI - [Treatment of angioma in children at the University Hospital Center of Vaud (CHUV), Lausanne]. PMID- 11887561 TI - [Occurrence of Trichophyton tonsurans tinea in Lausanne]. PMID- 11887562 TI - [Two new very promising antithrombotic agents: pentasaccharide and ximelagatran]. AB - The main anticoagulants presently used have several drawbacks. Two new compounds (pentasaccharide and ximelagatran) have shown in recent studies several interesting properties and results. Both are of non animal origin, do not induce thrombocytopenia and do not require laboratory controls. Pentasaccharide has a better efficacy than low molecular weight heparin in major orthopaedic interventions; it will be registered soon in several countries. Ximelagatran, which can be given orally and is in phase III investigation, could replace vitamin K antagonist in the future. PMID- 11887563 TI - [Dermatology, melanoma]. PMID- 11887564 TI - [Research note: "On the progression of the incidence of hip fractures in the elderly"]. AB - We studied 39,896 hip fractures that occurred in Quebec during 10 years. We found an increase in the incidence of hip fracture during this period. Demographical or anthropologic factors (such as smoking, alcohol consumption, residential placement), seasonal variations or socioeconomic factors alone could not give a satisfactory explanation for such an increase of this incidence. The pandemic disease theory might on the other hand give some explanation as to this above findings in hip fractures, considering that the average level of health of the elderly in 1991 is different from that in 1981. We suggest a hip fracture watch to allow for a follow-up on the general health situation of the elderly. PMID- 11887565 TI - [Abdominal pregnancy: study of a series of 11 cases]. AB - OBJECTIVE: To report our experiment concerning abdominal pregnancy. MATERIAL AND METHODS: Retrospective study of 11 cases of abdominal pregnancies operated by laparotomy between August 1994 and July 1999. RESULTS: 36.3% of cases corresponded to a first pregnancy. Four pregnancies had attended beyond the term of viability. Two children were born alive. The ablation of the placenta was generally complete (10/11 cases) without major bleeding. CONCLUSION: Rare affection of difficult clinical diagnosis, the abdominal pregnancy presents a significant perinatal mortality. PMID- 11887566 TI - [Allergic seasonal rhinoconjunctivitis without indigenous pollen sensitization: the example of the Arizona cypress]. AB - A 37-year old man from Lithuania presented with the typical symptoms of allergic rhinoconjunctivitis that appeared in spring this and the year preceding presentation. Skin prick tests for aeroallergens indigenous in Switzerland were all negative. A detailed history revealed that the patient was working in a building of a United Nation's agency surrounded by a park with numerous cypresses of Arizona trees. A skin prick test for pollens from Cupressus arizona was strongly positive, supporting the diagnosis of allergic rhinoconjunctivitis to pollens from this tree. Allergy to pollens from C. arizona is a widely prevalent as winter pollinosis in subtropical and Mediterranean areas but is exceptional in the continental and alpine climates of Switzerland. Five cases of C. arizona pollinosis have been diagnosed in the last 3 years in Geneva. It is notoriously difficult to diagnose cypressus pollen allergy, mainly because of the poor quality of in vivo and in vitro available tests. Horticulture with non-indigenous plants is responsible for pollinosis from C. arizona in Switzerland. The specialists in countries in which pollinosis from C. arizona is not endemic, need to be aware of arboriculture with non-indigenous plants, as well as the possibility of allergy to C. arizona. PMID- 11887567 TI - [Degeneration of eugenics? On the involuntary sterilization in Romand Switzerland in the 20th century]. PMID- 11887569 TI - [What is the future of medical records? Thoughts about nominal data]. PMID- 11887570 TI - [Recommendations for a policy on how to save medical records at the university psychiatric department for adults in the canton of Vaud]. PMID- 11887568 TI - [Recommendations for the management of melanoma]. PMID- 11887571 TI - [Detection of the sentinel ganglion in melanoma]. PMID- 11887572 TI - [Update on hepatorenal syndrome]. AB - The hepatorenal syndrome is a form of renal failure occurring in patients with advanced liver disease. The diagnosis is based both on the demonstration of low GFR and exclusion of other common causes of renal failure that may occur in patients with cirrhosis. Orthotopic liver transplantation remains the only curative treatment for this poor outcome disease; other modalities such as vasopressin analogues, transjugular intrahepatic portosystemic shunt or renal replacement therapies may serve as a bridge to transplantation. This article reviews the pathophysiology, diagnosis and current treatment of hepatorenal syndrome. PMID- 11887573 TI - [Antibodies against human recombinant erythropoietin: an unusual cause of erythropoietin resistance]. AB - In a 70 year old man with primary glomerulonephritis, severe anemia occurred after 4 years on hemodialysis and rHu-EPO. The usual mechanisms of EPO-resistance were excluded. A bone marrow sample showed red all aplasia. No circulating EPO could be detected; the serum inhibited the growth of erythroid precursors in bone marrow cultures. Immunoprecipitation identified an IgG anti-EPO, still active against deglycosylated EPO, i.e. directed against the peptidic matrix. Its high neutralising capacity and the absence of any immune abnormality rule out an auto antibody. Anti-rHu EPO immunisation is a very rare occurrence, made severe by transfusion-dependence and the risk of hemosiderosis. An immuno-modulating treatment can therefore be justified. PMID- 11887574 TI - [Quality management in the hospital environment: ISO 9002 certification: our experience]. AB - The nephrology-dialysis department of the Havre's hospital has launched a project of certification ISO 9002 in 1996, based on reflections from the centers of dialysis Upper Normandy. The problems encountered were mainly the lack of ways, a bad documentary structure, false ideas on the quality and a bad perception from the client. With the help of a quality manager, il has enable the project to advance and finalized, obtaining the certification of activity. In june 2000 "Procedure of taking in charge all the patient's medical and para-medical cost in the center of the Nephrology-Dialysis service" of the GHH (Hospital Group of the Havre) has brought not only the certificate but notable improvements on the level of documentary management, the errors, relationship clients-suppliers and projects of collaboration with the other services of nephrology dialysis of France. PMID- 11887575 TI - [Evaluation of the management of new kidney failure patients in Reunion (Do what I say, but don't do what I do!)]. AB - The aim of this study was to describe the new patients with chronic renal failure (CRF), from a nephrology unit, during a 1-year period. This analysis of recruitment allowed us to describe the "preventive treatments" initiate before and after the first contact with a nephrology unit. POPULATION AND METHODS: The medical records from patients coming for the first time for consultation or hospitalisation from 1.1.98 to 31.12.98 and with CRF (creatinine for men > 120 mumol/l, creatinine for women > 95 mumol/l) were retrospectively analysed. RESULTS: 98 patients (54 women and 44 men), among 248 new records (among a total of 2724 consultations and 948 hospitalizations) had a CRF. 76 patients (78%) had already a estimated clearance < 30 ml/min. 50% of the patients were sent from a general practitioner; 50% were diabetics; 30% were sent for a CRF discovered in a systematic evaluation. 26 patients arrived with at least one "preventive treatment". After their visit in nephrology, 48 more patients benefited from at least one "preventive treatment". 16 patients had no recommendations. 13 patients were hospitalised after the first consultation. 21 patients were put on dialysis immediately, 7 more in the first year, 7 died during the first hospitalisation. CONCLUSION: Patients with CRF are coming too late in nephrology (median creatinine 300 mumol/l), with a poor prognosis (7% death and 29% dialysis in 1 year) with no "preventive treatment" except symptomatic (hypertension). Furthermore, a big effort has to be done by the nephrologists to initiate the "preventive treatment" from the very first contact with patients with CRF. PMID- 11887576 TI - [Erythropoietin resistance]. PMID- 11887577 TI - [Quality management in medicine]. PMID- 11887578 TI - Endoscopic endonasal transsphenoidal surgery. PMID- 11887579 TI - Extracranial vertebral artery anatomy and surgery. PMID- 11887580 TI - Neurosurgical management of pineal tumours. PMID- 11887581 TI - Diffuse axonal injury after head trauma. A review. PMID- 11887582 TI - Neurobiology of epileptogenesis in the temporal lobe. PMID- 11887583 TI - Multi-modal monitoring of acute brain injury. AB - OBJECTIVE: To review the scientific basis for and utility of the traditional cerebral monitors used currently in neurointensive care, together with research techniques that are soon likely to become used in managing severe head injury and subarachnoid haemorrhage. DESIGN AND CONTENT: Firstly, the pathophysiology of acute brain injury including cerebral haemodynamics, oxygen and metabolism and the role of secondary insults are discussed. Secondly, the importance of assessment of cerebrovascular autoregulation and reactivity is reviewed together with methods for its continuous non-invasive measurement using transcranial Doppler and intracranial pressure/arterial pressure recordings. Thirdly, the respective roles of jugular venous oxygen and brain tissue oxygen monitoring are analysed. Fourthly, the use of cerebral microdialysis is described, together with an overview of its utility. CONCLUSION: Cerebral multimodal monitoring can be helpful for the optimal management of acute brain injury and essential for future exploratory trials of neuroprotective drugs. PMID- 11887584 TI - [Facial type, dental form and cosmetics]. AB - Is there a disgrace where there is no matching of the facial shape with the shape of the teeth? How can the dentist and the beautician solve it? This study proposes to: define the different facial shapes in Africans, specify the different forms encountered, propose a make-up according to these shapes. PMID- 11887585 TI - The antimicrobial effects of seven different types of Asian chewing sticks. AB - There are various plants, which are used as chewing sticks in different parts of the world. Several studies have been reported on the antimicrobial effects of chewing sticks on oral bacteria. The aim of this study was to compare the antimicrobial effect of aqueous extract of seven different types of chewing sticks found in Pakistan and other Asian countries. The ditch plate method was used to test the antimicrobial activity of seven Asian chewing sticks. It was found that at there was antimicrobial effect on Streptococcus fecalis at 50% concentration of Kikar (Acacia arabica) from Pakistan and Arak (Salvadora persica) from Saudi Arabia. The inhibition zones up to 2 mm were found in those two chewing stick extracts. It is recommended that the chewing sticks will be a great help in developing countries with financial constraints and limited oral health care facilities for their populations. PMID- 11887587 TI - [Noma and HIV infection: apropos of a case at the National Hospital Center in Bobo-Dioulasso (Burkina Faso)]. AB - Noma (Cancrum oris) is a gangrenous stomatitis arising from a periodontal infection and leading to severe soft tissue and bone destruction. The pathology involves numerous factors including local thrombosis, vascularitis, necrotizing gingivitis, immunodeficiency, gram negative and anaerobic infection. It is usually a disease of infants and malnourished children in tropical areas often occurring after a debilitating disease like measles. Recently, cases have been reported in adults especially elderly patients or during immunodeficiency states. Reconstructive surgery is often necessary to deal with destruction and sequel but is rarely accessible in developing countries. We report one case of noma (cancrum oris) in an HIV seropositive patient at the National Hospital in Bobo-Dioulasso. The noma was inaugural of AIDS in a 40 years old labourer coming back from Ivory Coast and no major opportunistic infection was associated. The course was fulminant leading to extensive facial gangrene with recurrent bacterial infections. The disease was fatal in this depressive, malnourished and diarrhoeic patient despite local surgical treatment, prolonged antibiotherapy and supportive care. Pathogenic mechanisms, management and preventive issues are discussed. PMID- 11887588 TI - [Necrotizing osteitis of the mandibular angle with facial asymmetry due to tooth extraction--apropos of a case]. AB - Five years after a tooth extraction, a bone necrosis of the mandibular angle happened, without consideration to the antibiotics. Such complication is rare. Living in poor socio-cultural and economic conditions, the patient tried to find help near tradipractionners, prayers groups, without any result. The treatment consisted to an extra oral incision, helping to eliminate bone sequestra, and the liquid around after a dynamic bimaxillary linkage by resin links was settled. About aetiology, only the questioning allowed to say that extraction was traumatic, putting in front of the surgical responsibility and post surgical survey. PMID- 11887586 TI - [Noma in children in a hospital environment in Bobo-Dioulasso: epidemiologic, clinical and management aspects]. AB - A retrospective study covering ten years (1987-1996) was conducted to assess the epidemiology, clinical features and management of cancrum oris (noma) in children from Burkina Faso. Fifty nine (59) children were admitted with cancrum oris at the paediatrics and maxillo-facial surgery units of Bobo-Dioulasso, the second town of Burkina Faso. The hospital prevalence of noma is 1.5/1000. 81% of the cases were in the 1 to 5 years age group and 58% were females. Predisposing factors include poverty, lack of immunization, malnutrition, bad oral hygiene, measles and parasitic diseases. The cheek was involved in 31% of the cases. Cure was obtained in 80% of patients after medical and surgical treatment. However, many sequels were observed. Post operative outcome is complicated by the children's growth and often results in retractions, recurrence of ulcers or constriction. Psychological and social problems are associated. Management is difficult in our setting because of the lack of information, cost of the treatment and the absence of well-equipped plastic surgery units. PMID- 11887589 TI - Massive osteolysis of the maxillo-facial bones: case report and review of the literature. AB - An unusual case of massive osteolysis destroying the left side of the mandible, the maxilla, the orbit, the cranium bones and the upper cervical spine is reported. The evolution of this disease was observed over a period of 4 years. The literature was reviewed, only 39 cases have been found involving the maxillo facial bones. In our case, attempts at surgery was without success. However, further radiotherapy with 35 Gy controlled the progression of this osteolysis. PMID- 11887590 TI - Setting up of an Odonto-Stomatology Department in the Faculty of Medicine in Cameroon. PMID- 11887591 TI - [Comparison of the clinical detection and the biological detection of dental caries]. AB - Different methods can be used to detect dental caries. The comparison of the clinical detection by the use of explorer and mirror to the biological detection by the measurement of alkaline phosphatase activity (ALP) in 107 men of 19 to 25 years old was performed. Biological detection reveals more active caries(APL activity > 116 Ul/L) than the clinical one. Absence of caries activity is expressed by an ALP activity between 86 and 116 Ul/L. The Chi 2 test shows that alkaline phosphatase is a statistically significant marker of dental caries and more it reveals its carious activity. PMID- 11887592 TI - [Dental caries in the public primary schools dependent on the Nabil Choucair Health Center in Dakar, Senegal]. AB - It is a holy study of observation and prevalence concerning a population of 268 pupils in first degree of the primary schools depending on the health center Nabil Choucair. 82% of these pupils have dental decay with DMFT egals 3.94. Our results suggest needs of preventive intervention on the intention to reach the objectives health for all by 2000 relatively to dental health in Senegal. PMID- 11887593 TI - Gums made simple. PMID- 11887594 TI - Gingival inflammatory response induced by chemical retraction agents in beagle dogs. AB - PURPOSE: The aim of this in vivo study on dogs was to investigate and compare the inflammatory potential of four different retraction agents on the gingival connective tissue. MATERIALS AND METHODS: All procedures on eight beagle dogs were performed under general anesthesia: taking oral hygiene measures, placing retraction cords medicated with four chemical agents into the gingival sulci, and taking tissue biopsies. The specimens were evaluated after a 10-minute exposure to chemical agents. The inflammatory response of the connective tissue underlying the sulcular and junctional epithelium triggered by retraction agents was assessed quantitatively. Microscopic images of tissue specimens were morphometrically analyzed using a computer-assisted morphometric method. RESULTS: The most intense inflammatory response in the connective tissue underlying the sulcular epithelium was triggered by astringent retraction agents--Racestyptine in specimens taken after 1 day and 1 week and Rastringent after 1 day (P < .05). Tetrahydrozoline-sympathomimetic vasoconstrictor (Visine) was found to have the lowest inflammatory potential. Retraction chemicals produced no significant effects on the connective tissue subjacent to the junctional epithelium. The ratio of the connective tissue area to that of the inflammatory infiltrate showed that 25% aluminum chloride (Racestyptine) was the most aggressive and tetrahydrozoline the least aggressive retraction agent used. CONCLUSION: All the retraction chemicals tested increased the infiltration with inflammatory cells in gingival connective tissue. PMID- 11887595 TI - Measurement of masticatory forces and implant loads: a methodologic clinical study. AB - PURPOSE: The aim of this study was to measure vertical masticatory forces in vivo using a method that should be insensitive to the location of bite force impact. MATERIALS AND METHODS: Two exchangeable implant abutments were equipped with strain gauges. In nine patients, the abutments were attached to implants supporting three-unit fixed partial dentures (FPD) in one mandibular chewing center. The signals of the two abutments were summed to give a force reading that was independent of the location of force impact along the FPD. In two subjects, an additional strain gauge was fixed under the pontic. With both setups, masticatory forces were measured in chewing of winegum. RESULTS: Total masticatory force displayed by the sum signal proved to be independent of the site of force application. Pontic strain gauges indicated only 42% or 84% of the force measured simultaneously by the corresponding sum signal of the abutments. In all nine patients, a mean total masticatory force of 220 N, with a maximum of 450 N, was found. The single abutments experienced mean loads of 91 N (anterior) and 129 N (posterior), with a maximum of 314 N. CONCLUSION: Measuring chewing force via bending of a pontic involves the risk of underestimation. Masticatory forces obtained with a method that was insensitive to the site of force application were higher than forces found with some other setups. PMID- 11887596 TI - Dentist-patient communication and patient satisfaction in prosthetic dentistry. AB - PURPOSE: Dentist-patient verbal communication dimensions on patient satisfaction were investigated in a prosthodontic context, controlling for the age and gender of patients and dentists and the amount of delivered prosthodontic treatment. Two concepts of satisfaction were defined, one for the single visit (satisfaction with care), and one for the overall result (satisfaction with treatment outcome). MATERIALS AND METHODS: Audio recordings of 61 patients meeting 15 dentists were made in three specialist clinics of prosthetic dentistry. The prosthodontic treatment periods with fixed tooth- or implant-supported prostheses, on average 20 months, were monitored by questionnaires. One visit near the end of each treatment period was audio recorded. The recorded verbal communication was analyzed with the Roter Interaction Analysis System-Dental. RESULTS: Bivariate analysis showed that patients of female dentists were more satisfied in the long term perspective than patients of male dentists. In logistic multivariate regression models, the verbal communication dimensions "information-dentist horizon" and "information-patient horizon," together with the mouth involvement of the prosthodontics, influenced patient satisfaction with treatment outcome. CONCLUSION: Patients undergoing extensive prosthodontic rehabilitation should be given the opportunity to ask and talk about their dental health, and dentists should minimize their question-asking and orientating behavior during the encounters to help improve patient satisfaction with treatment outcome. PMID- 11887597 TI - Changes in prosthetic screw stability because of misfit of implant-supported prostheses. AB - PURPOSE: The effect of two levels of prosthesis misfit on prosthetic screw stability was evaluated. MATERIALS AND METHODS: Two levels of vertical discrepancies--100 and 175 microns--were introduced between an implant-supported complete denture and the terminal abutment. An implant-supported complete denture without vertical discrepancy served as a control. Cyclic load was delivered vertically on the cantilever portion of the prosthesis next to the terminal abutment for 48 hours for each trial. A total of seven sets of new screws were tested for each level of fit. RESULTS: The results revealed significant prosthetic screw instability at both the 100- and 175-micron levels of discrepancy. CONCLUSION: Vertical discrepancies of 100 and 175 microns introduced between an implant-supported fixed complete denture and the terminal abutment resulted in significant prosthetic screw instability. PMID- 11887598 TI - A study of the influence of occlusal factors and parafunctional habits on the prevalence of signs and symptoms of TMD. AB - PURPOSE: The purpose of this study was to investigate the prevalence of the clinical signs and symptoms of temporomandibular disorders (TMD) and the relationship between occlusal factors, parafunctional habits, and TMD in a young adult nonpatient population. MATERIALS AND METHODS: A questionnaire including data from a history and clinical functional examination was used in the study. All 230 subjects were male recruits, from 19 to 28 years of age (mean 21.3 years). RESULTS: Thirty-eight percent of the subjects reported at least one symptom, while in 45% of the subjects at least one sign of TMD was recorded. Temporomandibular joint clicking (40%) and pain on palpation (34%) were the most commonly recorded signs. Multivariate logistic regression analysis showed several weak but statistically significant correlations between the occlusal factors, parafunctional habits, and TMD in this nonpatient population. TMD signs were thus weakly correlated with malocclusion traits (angle Classes II/1, II/2, III, and cross bite), interferences in retruded contact position, midline discrepancy > or = 2 mm, < or = 10 contacts during maximal biting pressure, nonworking-side interferences, horizontal overlap > or = 5 mm, and parafunctional habits (teeth clenching and teeth grinding). CONCLUSION: Some association between occlusal factors and TMD signs was found. However, this association cannot be considered unique or dominant in defining subjects with TMD in the population. PMID- 11887599 TI - Factors influencing the clinical composite assessment of denture-supporting tissues. AB - PURPOSE: This study attempted to (1) clarify the factors influencing the clinical composite assessment of denture-supporting tissues in edentulous patients by experienced prosthodontists and (2) develop a method for quantitative assessment of supporting tissue. MATERIALS AND METHODS: Seven three- or four-grade scales for clinical factors were used to evaluate supporting tissue in 317 complete denture patients. The general assessment of supporting tissue was conducted with a 10-point scale. The contribution of each grade of the seven factors to the general assessment level was determined by a multiple regression analysis. RESULTS: Based on the contributions of the seven factors, a method for quantitative assessment of supporting tissue was established, category scores for these factors were calculated, and the sums of the category scores were converted to an integer between 0 and 100. The resulting quantification score was closely correlated with the general assessment of the supporting tissue by experienced prosthodontists. CONCLUSION: The factors influencing the clinical composite assessment of denture-supporting tissues in edentulous patients by experienced prosthodontists were clarified. The quantitative assessment method developed in this study is simple and might be used for preoperative diagnosis and treatment planning. PMID- 11887600 TI - Some variables influencing the bond strength between PMMA and a silicone denture lining material. AB - PURPOSE: The purpose of the present study was to investigate the effect of roughening the denture base surface on the tensile and shear bond strengths of a poly(dimethylsiloxane) resilient lining material (Molloplast-B) bonded to a heat cured acrylic resin denture base material. These measured bond strengths were compared to those obtained by packing the soft lining material against poly(methyl methacrylate) (PMMA) denture base acrylic resin dough. MATERIALS AND METHODS: Three groups of 10 specimens each were constructed for both tensile and shear tests. In the first group, Molloplast-B was packed against cured PMMA denture base surface. In the second group, Molloplast-B was packed against cured PMMA denture base whose surface had been roughened with an acrylic bur. In a third group, Molloplast-B was packed against PMMA denture base acrylic resin dough. RESULTS: Molloplast-B exhibited significantly higher tensile and shear bond strengths when packed against acrylic resin dough. Roughening the denture base surface prior to the application of Molloplast-B had a statistically significant weakening effect on tensile bond strength compared with the smooth surface and the acrylic resin dough. For the shear bond strength, roughening the surface produced a nonsignificant increase compared with the smooth surface, but the bond was weaker than when packed against acrylic resin dough. CONCLUSION: Significant differences in tensile and shear bond strength were recorded between the three methods used to bond Molloplast-B to denture base material. PMID- 11887602 TI - A review of clinical and technical considerations for fixed and removable implant prostheses in the edentulous mandible. AB - PURPOSE: The aim of the present article is to review some of the technical treatment options for implant prostheses restoring the edentulous mandible, mainly based on the Branemark system. MATERIALS AND METHODS: Clinical and technical aspects are discussed for the three established concepts: (1) implant supported fixed prosthesis, (2) removable implant-supported overdenture, and (3) combined implant-retained and soft tissue-supported overdenture prosthesis. RESULTS: The framework of an implant-supported fixed screw-retained prosthesis can be processed in gold, Co-Cr alloy, or titanium with casting, laser-welding, or milling techniques. To improve the stability and retention of a conventional complete denture, one to four implants are indicated, and unsplinted (single attachments) or splinted designs (bar systems) can be applied. The design of the overdenture prosthesis must be carefully planned according to the requirements to ensure adequate stability and optimal form, contour, and esthetics, and the patient's best comfort. CONCLUSION: A large variety of different treatment modalities exist for both the fixed and removable mandibular implant prosthesis. Clinical and technical aspects should be considered at the beginning of the treatment to: (1) select the optimal implant position, (2) establish an adequate number of functional units, (3) select the appropriate retainers, and (4) apply the best technique for framework processing and veneering. PMID- 11887601 TI - Comparison of marginal integrity of ceramic and composite veneer restorations luted with two different resin agents: an in vitro study. AB - PURPOSE: The aim of the present study was to examine the marginal integrity of a composite veneer, Artglass, in comparison to a ceramic veneer, IPS Empress, by using two different luting agents. The study also aimed to evaluate the effect of grit blasting on marginal integrity of IPS Empress ceramic veneers. MATERIALS AND METHODS: A total of 42 human maxillary incisors were used to construct 14 veneers for each experimental group: (1) Artglass veneers, (2) IPS Empress veneers with grit blasting, and (3) IPS Empress veneers without grit blasting. Veneers were luted with Variolink II High Viscosity and Variolink Ultra. The restored teeth were sectioned buccolingually and mesiodistally, and marginal gap width measurements were made at 200x magnification. The data were statistically analyzed with analysis of variance. RESULTS: The mean marginal gap width varied from 105 to 182 microns. No significant differences were recorded between the marginal gap widths in relation to different types of veneers or luting agents. However, significantly higher marginal discrepancies were observed for the incisal margins of IPS Empress veneers without grit blasting. CONCLUSION: In view of the higher marginal discrepancy of IPS Empress veneers divested without grit blasting, the use of this alternative technique was not beneficial. The use of a highly filled resin luting agent (Variolink Ultra) did not cause an increase in marginal gap widths of the veneers. PMID- 11887603 TI - Color comparison of two shade guides. AB - PURPOSE: The aim of this study was to analyze color parameters and color compatibility of two randomly chosen Vita shade guides, as well as to propose possible clinical guidelines. MATERIALS AND METHODS: Data were recorded using a colorimeter set to standard illuminant source C and the CIE L*a*b* system. A custom adapter system, which allowed a measuring area at the middle third of the tabs, was produced. Each of 42 tabs was recorded one time each on three different days. Color distribution was examined in diagrams whose coordinates were L*a*b* and L*C*H degree color coordinate pairs. Color coordinate ranges and coverage error were examined using the corresponding equations and statistical methods. RESULTS: The method repeatability was approximately delta E* = 0.1. Color difference ranges of Vitapan Classical and Vitapan 3D Master were 14.3 and 19.2, respectively. Color coordinate ranges of Vitapan Classical were as follows: delta L* = 12.8; delta a* = 1.7; delta b* = 9.0; delta C* = 9.0; and delta H degree = 7.4. Corresponding values for Vitapan 3D Master were delta L* = 15.3; delta a* = 3.4; delta b* = 16.3; delta C* = 16.6; and delta H degree = 10.5. Coverage error of Vitapan 3D Master to Vitapan Classical was 1.4 +/- 0.6, while vice versa it was 2.0 +/- 1.5. CONCLUSION: Compared to Vitapan Classical, chromaticity ranges of Vitapan 3D Master were extended in the desired directions: hue was extended toward yellow-red, and saturation was extended toward more saturated tabs. Compared to Vitapan Classical, Vitapan 3D Master tabs were more uniformly spaced. The examined shade guides were found to be color compatible. PMID- 11887604 TI - A 3-year clinical evaluation of Cerana prefabricated ceramic inlays. AB - PURPOSE: The present study was carried out to evaluate Cerana Class I and II tunnel preparations in combination with Cerana inlays. MATERIALS AND METHODS: Sixty-eight patients were treated with 89 Cerana restorations, 53 in the maxilla and 36 in the mandible. Of these, 47 were Class I and 42 were Class II. The restorations were examined after 1, 2, and 3 years by the author together with the treating clinician. for the evaluation, the California Dental Association quality evaluation system was used. RESULTS: At the 3-year recall, 59 (66%) restorations were examined. One restoration had been exchanged after 1 week, and 10 (11%) restorations had fractured before the 3-year follow-up. A total of 19% of the marginal ridges had fractured over the 3 years. These fractures were in connection with Class II tunnel preparations, except for one Class I filling. All of the Cerana inlays had acceptable marginal adaptation. Marginal discoloration was seen around 8% of the restorations. The color match was excellent, and the majority of the restorations had a smooth surface. Caries was seen in five teeth, three in connection with fractured marginal ridges. CONCLUSION: The results indicate that Cerana is an alternative to composite resin restorations in Class I situations, but should be avoided in connection with Class II tunnel preparations. PMID- 11887605 TI - A study on the fracture strength of implant-supported restorations using milled ceramic abutments and all-ceramic crowns. AB - PURPOSE: The purpose of this study was to compare five different abutment-crown combinations for single implant-supported restorations regarding their capabilities to withstand loads. MATERIALS AND METHODS: Fifty implants were placed into resin blocks, and the restorations were connected to the implants. The five tested restorations were: (1) metal-ceramic crowns cemented to titanium abutments, (2) In-Ceram crowns cemented to titanium abutments, (3) Celay feldspathic crowns cemented to titanium abutments, (4) In-Ceram crowns cemented to milled ceramic abutments, and (5) Celay feldspathic crowns cemented to milled ceramic abutments. The specimens were loaded at 0- and 45-degree angles to the long axis, and the load values at the moment of failure were recorded using a universal testing machine. RESULTS: The fracture strengths under vertical loading were greater than those under oblique loading. The fracture strengths of metal ceramic crowns cemented to the titanium abutments were higher than those of all ceramic crowns cemented on the milled ceramic abutments, regardless of loading direction. There were no differences in the fracture strengths of the ceramic crowns between the two different abutment types under oblique loading. CONCLUSION: All-ceramic crowns on the milled ceramic abutments were weaker than the metal-ceramic crowns on the titanium abutments under oblique loading. PMID- 11887606 TI - Combatting the assault on childhood mental disorders. PMID- 11887607 TI - An ethical dilemma: clinical psychologists prescribing psychotropic medications. AB - The use of psychotropic medication to treat psychiatric disorders has surged in recent years, and while commonly prescribed, the question of who should be allowed to prescribe such medication has become an increasingly important issue to nurses. Psychologists have historically functioned in roles such as psychotherapy and psychological testing, but as standards of care for psychiatric disorders incorporate medication, reimbursement for psychotherapy is declining. Medication prescription and management have not been traditionally seen as the role of the psychologist, however, many clinical psychologists have begun to advocate for prescription authority as a legally sanctioned role for their profession. This article addresses the issues of clinical psychologists seeking prescriptive privilege. It will be argued that the current paradigm of psychology rejects the neurobiological basis of mental illness and that psychologists prescribing medication presents an ethical dilemma for nurses. It is the contention of the author that nurses have an ethical responsibility to advocate against the extension of the psychologist's role into the prescription of medications. PMID- 11887608 TI - Evaluation of training designed to prevent and manage patient violence. AB - This study was designed to determine whether the test responses of mental health care workers (n = 118) showed significant improvement after attending a training session about managing violence. Four variables (knowledge, attitude, self efficacy, and behavioral intention) were measured before and after staff attended a training program that consisted of two commercial programs: the Nonviolent Crisis Intervention (CPI) and Handle with Care. The Nonviolent Crisis Intervention is designed to teach staff how to prevent and control disruptive behavior of clients. "Handle with Care" is a combination of lecture and demonstration of self-defense skills and restraining methods for staff who work with potentially assaultive patients. The research team used a one group, pretest/posttest study design for the evaluation. The study location was an acute care psychiatric hospital located in the southwestern United States. Hospital staff completed a pretest, participated in a 12-hour intervention, and completed a posttest immediately after the intervention. The evaluation of staff responses demonstrated improvements in posttest scores that were significant for knowledge, attitude, self-efficacy, and behavioral intention to use the training techniques. More research is needed regarding evaluation of programs that train mental health care workers to prevent and manage patient violence. Such research can help us develop more effective programs. PMID- 11887609 TI - Infertility and the role of psychotherapy. AB - Infertility affects 6 million American women and their partners, about 10% of the reproductive age population. Recent improvements in medication, microsurgery, and assisted reproductive technology (ART) make pregnancy possible for more than half the couples pursuing treatment, yet infertility is more than a medical condition. Infertility touches all aspects of a person's life. It affects how individuals feel about themselves, their relationships, and their life perspective. Stress is only one of a myriad of emotional realities that couples facing infertility deal with, often for extended periods of time. In addition to ongoing stress, infertility creates issues of guilt, anxiety, tension within the relationship, and feelings of depression and isolation. Treating couples and individuals who are involved in the journey of dealing with infertility is an opportunity for clinical specialists in psychiatric mental health nursing. The expertise of clinical specialists both in providing psychotherapy services as well as providing a bridge of understanding of sophisticated medical and surgical procedures places them in a unique position among the various disciplines offering mental health services. This paper provides an overview of the psychological issues that play a role in psychotherapy with this population and suggests specific clinical interventions and potential role expansion for psychiatric clinical specialists. PMID- 11887610 TI - Evaluating the role of a Psychiatric Consultation-Liaison Nurse in the Australian General Hospital. AB - The importance of Psychiatric Consultation-Liaison Nursing (PCLN) in improving health outcomes for patients experiencing mental health problems has received some attention in the nursing literature. However, little effort has been made to evaluate the impact of this role. The study presented in this paper was conducted in order to assist in redressing this paucity. Focus groups were conducted with nurses (n = 17) employed at a large general hospital in Melbourne, Australia. The nurse participants were asked to discuss their views about utilizing the services of the specialist PCLN employed in this hospital. The responses of participants were overwhelmingly positive. The four main themes to emerge from the study were: helping staff, making contact, implementing strategies, and utilizing attributes. These findings are presented and discussed as a contribution to and extension of the existing literature. PMID- 11887611 TI - Nurses' views on situations related to privacy in providing home care for persons with long-term mental illness: an exploratory study. AB - The main aim of this exploratory study was to explore the views of nurses and mental health care workers on situations related to patient privacy in the home care of persons with long-term mental illness in Sweden. A specifically designed questionnaire was developed from theoretical constructs obtained in a qualitative study and distributed to 1,139 respondents. Data from 660 district nurses, psychiatric nurses, and mental health care workers revealed significant differences in perceptions by age, gender, and professional groups. For example, psychiatric nurses and mental health care workers indicated to a greater extent than district nurses feelings that relate to intrusion on patient privacy. Respondents who were 41 years or younger also indicated to a greater extent than those who were older that they felt home care was an intrusion on patient privacy. Moreover, men indicated to a greater extent than women feelings of insecurity in their professional role. Further investigation is needed, especially into how nurses deal with situations that intrude on patient privacy and how nursing intervention impacts on the patients' own sense of privacy. PMID- 11887612 TI - The practice of honor crimes: a glimpse of domestic violence in the Arab world. AB - Domestic violence, especially violence against women, is a serious health problem in the United States and in many countries of the world. However, information on violence against women in the Arab culture is scarce. The purpose of this descriptive study is to investigate the incidence of violence against women in one Middle Eastern country. The focus of the research is to determine the cultural context in which violent crimes against women are committed and the social and legal implications of such crimes. The research method included: (1) a review of all court files of women murdered during 1995 in the country of Jordan and, (2) the social norms and sanctions against persons who commit crimes against women. Of 89 homicide cases reviewed, 38 involved female victims. Analysis of the court files of the 38 murdered women indicated that a male relative of the female victim, primarily the brother, committed the majority of the murders. The most common cause for the murders provided in the files was "honor crime." Honor crime was defined as crime committed against women by their male family members because the women had violated the honor of their family. Cultural norms and practices including the legal practices related to honor crimes support the practice of killing women for sexual misconduct and excuse perpetrators of the crimes from punishment. PMID- 11887613 TI - Effect of colostrum intake on diarrhoea incidence in new-born calves. AB - In a survey which lasted one year and included data of 73 dairy cows with their calves, colostrum immunoglobulin G (IgG) of 22 primiparous cows and serum IgG of their calves were lower than the corresponding IgG levels of 51 multiparous cows and their calves. Serum IgG concentration was not correlated with diarrhoea incidence. Although there were no seasonal differences in the IgG concentration of colostrum and calf serum, neonatal diarrhoea incidence was higher in calves which were born in winter than in calves which were born in summer (P < 0.01). Thus the high diarrhoea incidence in winter was not a consequence of an insufficient IgG transfer to the calves. The 60 calves of the second study were fed colostrum on the first day of life. From the second to the tenth day 28 experimental calves received milk and 0.5 l of surplus colostrum of the first and second milking twice a day, whereas 32 control calves received milk only twice a day. Two of the 28 experimental and 11 of the 32 control calves suffered from diarrhoea during the first ten days of life (P < 0.05). These results show that the ingestion of surplus colostrum in addition to milk after the first day of life protects the new-born calf against infectious diarrhoea. PMID- 11887614 TI - [Intracranial astrocytomas in eight cats: clinical and pathological findings]. AB - Intracranial astrocytomas are rarely diagnosed in cats. Clinical and pathological aspects of these tumors are more often described in humans and dogs. The classification scheme used in human medicine is of important prognostic value. We have analyzed clinical neurological and pathological findings from 8 cats with intracranial astrocytomas. The animals were 10.1 years old in average and presented with a history of tetraparesis (n = 3), epilepsy (n = 2), loss of balance (n = 3) and dyspnoe (n = 1). The latter cat died immediately after the first presentation while the other animals were euthanized because of a progressive course of the symptoms despite therapy. Even though feline astrocytomas, that we could classify into 4 different types in this study, are clinically and pathologically well correlated with those of other species, a prognostically useful classification has never been established before. PMID- 11887615 TI - [Pseudohypoadrenocorticism in two Siberian huskies with gastrointestinal parasitoses]. AB - Two Siberian Huskies were presented because of anorexia, chronic diarrhoea, polydipsia and polyuria. The most significant clinical finding was a severe dehydration. Laboratory results showed severe electrolyte disturbances with sodium: potassium ratios of 13.2 and 15.4, respectively. Hypoadrenocorticism could be excluded in both dogs with an ACTH stimulation test. The plasma aldosterone concentrations before and after stimulation were very high, which might be due to a stimulation by the electrolyte disturbances. The final diagnosis was an infection with the whipworm Trichuris vulpis and secondary pseudohypoadrenocorticism. A life-threatening dehydration syndrome with hyponatremia, hyperkalemia and metabolic acidosis due to intestinal parasitism has been reported in earlier studies. Up till now sodium: potassium ratios less than 14, however, have only been seen in animals with Morbus Addison. The severe hyperkalemia in these dogs might be due to a particular sensitivity in this breed. PMID- 11887616 TI - [Schistosoma reflexum in a female bovine fetus with synaptonemal complex abnormalities]. AB - In this study we present mitotic- and meiotic investigations in an anatomical abnormal bovine fetus. The abnormality could be classified as "schistosoma reflexum", which was never described in fetuses in the literature. In the mitotical chromosome preparations from fibroblast cultures the examined fetus showed no chromosomal difference in comparison to the standard synaptonemal complexes (SC) which were prepared from the fetus at the age of 92 days post coitum. In the SCs from the abnormal fetus 43.75% of the investigated cells showed abnormalities such as "loop," "nonhomologue pairing" and "multivalency" in the pairing behavior of the chromosomes. In comparison, less than 5% of the cells in normal embryos showed such abnormalities. PMID- 11887617 TI - [Changing perception of hereditary eye diseases]. AB - The authors present the cases of two parents with Usher syndrome type I who appeared to have normal offspring, and two families, one with autosomal dominant retinoblastoma and a RB1-gene mutation and one with primary open angle glaucoma and a myocilin gene mutation, in whom DNA-analysis was used to see whether check ups were needed. The field of ophthalmogenetics comprises many disorders, both congenital and those with a later onset. Mendelian, mitochondrial, as well as multifactorial heredity is seen. Recent progress in this field, especially in molecular genetics, has created new possibilities, but some situations appear to be more complex than previously assumed. Particularly if there is genetic heterogeneity or multifactorial inheritance, possibilities for counselling and DNA analysis remain limited. PMID- 11887618 TI - [The electric heart assist device for patients on the waiting list for a heart transplant: a bridge too far]. AB - Left-ventricular assist devices are being increasingly used as a bridge to heart transplantation in patients with end-stage heart failure. However, current evidence does not support their widespread application for this indication. Other indications such as cardiogenic shock in acute myocardial infarction, or severe myocarditis may possibly be more fruitful. The use of these devices in the chronic support of patients with end-stage heart failure has recently been evaluated in patients excluded for heart transplantation. Patients treated with an assist device had a better quality of life than patients on drug treatment, and a greater number of them were still alive after two years (23% versus 8%). In the future, novel options such as skeletal muscle cell transplantation, may offer a feasible alternative to heart transplantation. PMID- 11887619 TI - [The revised Dutch College of General Practitioners' standard on COPD and the first international WHO standard: differences and similarities]. AB - Two standards on COPD have recently been published: the revised national standard from the Dutch College of General Practitioners and the first international standard published by the World Health Organization and the US National Heart, Lung and Blood Institute. The reduced emphasis on the role of spirometry in the monitoring and evaluation of treatment is an important change in these new standards compared to previous ones. Cessation of smoking is considered to be central to the prevention and treatment of COPD. Doctors should strongly support this approach and, more than before, are urged to view COPD as a disease caused by addiction. Bronchodilators are the cornerstone of symptomatic treatment of COPD, particularly the long-acting ones due to their ease of administration and effective treatment of morning dyspnoea. Inhalation corticosteroids should only be administered as a trial treatment and only under certain conditions. Continuation of treatment with these agents is only justified if there is a demonstrated improvement in lung function, exacerbations or symptoms, although the precise area of indication is not yet clear. PMID- 11887620 TI - [Negative effects of passive smoking on the (unborn) child]. AB - The negative effects of passive smoking on the health of the foetus or child continue to receive little attention, despite the large volume of research in this area. Passive smoking during pregnancy is associated with low birth weight, a reduction in head circumference at birth, and a far higher incidence of sudden infant death syndrome. Exposure to cigarette smoke also leads to a decreased lung function, an increased risk of severe infections, including respiratory syncytial virus bronchiolitis, meningococcal disease and middle ear infections. There is no association between passive smoking and the development of allergic asthma, but passive smoking does cause an increase in the prevalence of respiratory symptoms in children with or without asthma. Finally, there is a relation between passive smoking and behavioural disorders including attention deficit/hyperactivity disorder (ADHD). Passive smoking before birth seems even more harmful than after birth. A causal relationship is suggested in most studies, or has been proven by animal experiments. A decreased birth weight in general increases the risk of developing chronic diseases as an adult, such as hypertension, cardiovascular disease and type 2 diabetes mellitus. This extensive knowledge about the adverse health effects of smoke exposure in (unborn) children deserves greater attention in the counselling of pregnant women, and in anti-smoking campaigns. PMID- 11887621 TI - [Guidelines for follow-up of patients with prostate carcinoma]. AB - The follow-up investigation of patients with prostate cancer is mainly conducted on the basis of complaints, serum prostate-specific antigen and digital rectal examination. Other investigations, such as bone scintigraphy, are only performed on indication. The frequency and duration of the follow-up investigation, as well as the consequences of the findings during this, depend on the stage of the disease, the patient's wishes and the possibilities of additional treatment. PMID- 11887622 TI - [Diagnostic image (77). A women with retinal hemorrhage]. AB - In a 32-year-old pregnant woman with visus loss due to a macular bleeding ultrasonography and retinoscopy revealed an osteoma. PMID- 11887623 TI - [From gene to disease; the dystrophin gene involved in Duchenne and Becker muscular dystrophy]. AB - Duchenne and Becker muscular dystrophy (DMD and BMD) are progressive disorders, which almost exclusively affect males. DMD is the more severe type with an onset at 2-3 years of age. Patients become wheelchair-bound before the age of 13 and often die due to cardiac arrest or respiratory insufficiency. BMD, a more varying phenotype which may overlap with limb girdle muscular dystrophy (LGMD), has a less severe muscle weakness which starts later than in DMD patients. DMD carriers may show some muscle weakness. The dystrophin gene (2.4 Mb), known to be involved in DMD/BMD, codes for a 427 kilodalton muscle-specific protein named dystrophin as well as several tissue-specific isoforms. Dystrophin, as part of a membrane bound complex of proteins, connects the cytoskeleton of the muscle cell to the extracellular matrix. Since 1985, when highly reliable carrier detection and prenatal diagnosis at the DNA level became possible, over 250 prenatal tests have been performed. Molecular genetic analysis, highlighted a phenomenon called germinal mosaicism, which explains the recurrence of de novo mutations and led to the discovery of the so-called reading-frame rule, which helps to discriminate between DMD and BMD. Fifteen years after the discovery of the dystrophin gene, mutations can be detected in 95% of the patients, while the remaining 5% are still hiding within this very large gene. PMID- 11887624 TI - [Good results after closure of a arterial septal defect during heart catheterisation instead of surgery]. AB - OBJECTIVE: Evaluation of the first results in the Netherlands of percutaneous and transvenous closure of an ASD II in children with an Amplatzer Septal Occluder (ASO). DESIGN: Prospective. METHOD: Data were collected from children with an ASD II prior to, during and up to 24 months after the insertion of an ASO during heart catheterisation in Leiden University Hospital, the Netherlands. RESULTS: Between 1 January 1998 and 29 February 2000, 28 patients (12 girls, 16 boys; mean age: 74 months (range: 15-198 months)) underwent heart catheterisation to close an ASD II with an ASO. In 26 patients an ASO could be placed without significant complications. The size of the device varied from 9-34 mm (median 16 mm). In one patient ASD closure was not attempted because of multiple ASDs. In another patient the procedure was stopped after air embolism into the coronary arteries had occurred during preparation of ASO implantation. In 23/26 patients with an implanted ASO, no residual shunt was present after 24 hours. One child, in whom the defect was found to be closed after 24 hours and after three weeks, returned abroad and was lost to follow-up. After one year all defects (n = 22) were completely closed. CONCLUSION: Percutaneous transvenous closure of an ASD II with an ASO was possible, was not associated with any significant complications and had a high success rate, even in relatively young children with large defects. PMID- 11887625 TI - [The application of electric heart assist devices in 3 patients with end-stage heart failure as a bridge to transplantation]. AB - Left-ventricular assist devices have already gained an international place in the treatment of end-stage heart failure. It is expected that in future they will be increasingly used as a temporary bridging following the recovery from heart failure and to a lesser extent as a bridge to heart transplantation. Three patients with end-stage heart failure, men aged 68, 57 and 49 years, received a left-ventricular assist device (LVAD) as a bridge to transplantation. The device chosen was a Heartmate Vented Electric System (ThermoCardiosystems; Woburn, Massachusetts, US). In this system a pump is implanted under the diaphragm and connected to the apex of the left ventricle and the pars ascendens aortae. The first two patients reached the time of transplantation and used the LVAD for 367 and 416 days respectively. The third patient died after the pump had been implanted, due to progressive right-ventricle failure. The first patient died shortly after the heart transplantation. PMID- 11887626 TI - [Necrotising pancreatitis as an unexpected complication in seriously ill patients]. AB - Three patients, two men aged 70 and 73 years, respectively, who underwent surgery due to an abdominal aortic aneurysm, and a woman aged 75 years, who was operated on due to acute arterial embolic occlusion of both legs, developed abdominal complaints post-operatively. These were found to be caused by necrotising pancreatitis. The accompanying fluid accumulation was drained percutaneously. Two patients recovered; the 73-year-old man died suddenly, possibly as a result of burst aortic sutures. In patients with a serious condition, necrotising pancreatitis should be considered in the case of a generalised inflammatory reaction and abdominal symptoms. Percutaneous drainage of infected necrotic tissue can sometimes improve the patient's condition, making surgery possible at a later stage. PMID- 11887627 TI - [Perinatal pertussis: from mother to child]. PMID- 11887628 TI - [Perinatal pertussis: from mother to child]. PMID- 11887630 TI - [Total mesorectal excision and previous radiotherapy in patients with rectal carcinoma: good initial treatment outcome]. PMID- 11887631 TI - [High vacuum extraction: justifiably obsolete taking into account the risks for the child]. PMID- 11887633 TI - Treatment of malignant biliary stenosis: which stent to use? AB - The insertion of a biliary endoprosthesis has become standard therapy in the palliative treatment of a malignant biliary stenosis. For plastic stents, stent occlusion results from clogging caused by the adherence of proteins, bacteria, and sludge to the inner stent wall, resulting in a median stent patency of about 4 to 5 months. No major gain in stent patency can be obtained by the omission of side holes, nor by changes in stent material. Putting the stent inside the bile duct, in a suprapapillary position, does not lead to a longer stent patency. The prophylactic administration of antibiotic agents such as ciprofloxacin or norfloxacin, that are active against the gram-negative enterobacteriaceae leading to stent clogging, could have potential advantages but still needs further study. The insertion of a straight 10 French gauge polyethylene Amsterdam-type of prosthesis in a normal transpapillary position, and without the administration of any prophylactic antibiotic treatment, can still be regarded as state-of-the-art therapy with a plastic stent. This mainly holds true for those patients with a low life expectancy of only a few months, such as it is often the case in patients in a poor clinical condition, with liver metastasis, or with a large primary tumor. Patients with a longer life expectancy can be treated with a self expandable metallic stent. At present, there is no major indication that coated metallic stents will perform any better than the uncoated ones. PMID- 11887632 TI - Compassionate use of Gemzar in advanced pancreatic cancer: a Belgian experience. AB - Gemzar is a nucleoside analog that has been shown to be superior to 5 fluorouracil for the treatment of advanced pancreatic cancer in terms of both clinical benefit and survival. This open label program enrolled 214 patients. Patients eligible for this program had advanced or metastatic pancreatic cancer, received up to one previous chemotherapy, a baseline Karnofsky performance status (KPS) of at least 50, measurable or evaluable disease, adequate organ function defined as: absolute leucocyte count > 3 x 10(9)/L, platelet count > 100 x 10(9)/L, hemoglobin > 9 gr/dL, total bilirubin < 2 x upper limit of normal (ULN), creatinine < 2 x ULN, ALT and AST levels < 5 x ULN and were at least 18 years. A 1000 mg/m2 of Gemzar was administered weekly up to 7 weeks followed by a week of rest, then once weekly for 3 weeks out of every 4 weeks. The median age at inclusion was 64 years, 52% of the patients were male, 27% were 70 year or older, 66% had stage IV disease, 66% had a KPS of 80 or higher and 34% had received no prior chemotherapy. The overall response rate is 7%. A time-to-first-serious event analysis was performed since only a limited number of dates of death were available. The first serious event (FSE) was considered as the earliest of the following: increase by at least 2 of the pain score, deterioration of KPS of at least 20, documentation of progressive disease or death. The median time to FSE was 4 months, the free FSE rate at 1 year was 14%. We conclude that the results observed in this program confirm the established efficacy of Gemzar in pancreatic cancer. PMID- 11887634 TI - Recent insights into pathophysiology of sepsis-associated liver dysfunction. PMID- 11887635 TI - Ileo-caecal actinomycosis: report of a case simulating complicated inflammatory bowel disease. AB - Abdominal actinomycosis is a rare infectious disease caused by Actinomyces israelii, a gram-positive anaerobic saprophyte germ that is a normal inhabitant of the upper intestinal tract in humans. Actinomyces israelii rarely cause abdominal infections or actinomycosis. Abdominal actinomysosis is characterised by fistulae and abscesses and may mimic cancer or inflammatory bowel disease. Abdominal actinomycosis is difficult to diagnose preoperatively, and often require surgical removal of the diseased tissue, allowing pathologists for giving the definitive diagnosis, revealed by characteristic "sulfur granules". The authors report herein the case of a 47-year-old man who presented with diarrhoea and abdominal pain. Abdominal computed tomography evoked complicated inflammatory bowel disease and surgical procedure was decided. Laparoscopic exploration did not provide further significant information, and laparotomy with diseased bowel resection was performed. Pathology demonstrated "sulfur granules" and allowed the diagnosis of abdominal actinomycosis. This case demonstrated that abdominal actinomycosis should be included in the differential diagnosis when computed tomography shows an infiltrative and inflammatory mass. PMID- 11887629 TI - [Increased incidence in gonorrhea and Chlamydia trachomatis infection in a general practice in Amsterdam-Zuidoost, 1996-2000]. PMID- 11887636 TI - Multicentric tumor of the esophagus and its clinical significance; case report and review of the literature. AB - Simultaneous occurrence of the esophageal tumor at multiple sites in a single patient is unusual. Here in, we described a patient had three separate tumoral nodules with ulceration of the esophagus. Thorax CT scan, X-ray and endoscopy revealed the only two tumoral lesion at the lower esophagus. But, the other tumoral lesion at the upper part of the esophagus was detected at operation and histopathologic examination. The aspect of treatment was changed according to this new condition during the operation. We discussed the multicentric tumoral lesions of the esophagus in view of the literature. PMID- 11887637 TI - Molar control. Part 1. PMID- 11887638 TI - The A line. PMID- 11887639 TI - Do you attempt adult maxillary expansion? PMID- 11887640 TI - Maxillary canine transposition. PMID- 11887641 TI - Time-saving closing loops for anterior retraction. PMID- 11887642 TI - Philippe self-ligating lingual brackets. PMID- 11887643 TI - Elastic traction with Essix-based anchorage. PMID- 11887644 TI - Surebonding indirect lingual retainers. PMID- 11887645 TI - Stretching the limits of delegation. PMID- 11887646 TI - The midwife's role: challenges and changes in the post-medical movement towards woman-centred midwifery care. AB - The current movement towards woman-centred midwifery care is creating many changes for practicing midwives. In order to address these changes, midwives, particularly those working in the hospital system, need to be clear about their role in the care of childbearing women, and how they can be part of the solution in changing the fractured maternity care currently operating under the medical system. This article examines some of the issues arising for midwives, such as autonomous practice, use of obstetric language, and education for the new generation of midwives. PMID- 11887647 TI - Antenatal education classes in Victoria: what the women said. PMID- 11887648 TI - What mothers want: a postnatal survey. AB - Mothers in the public health care system undergo mandatory early discharge after childbirth. The challenges associated with the decreasing length of hospital stay have rarely been investigated from a service consumer perspective. The aim of this study was to identify mothers' needs in the immediate postpartum period. An inpatient survey of 500 postnatal mothers undertaking the Early Discharge Program. Survey questions aimed to elicit the needs of mothers in the immediate postnatal period; perceived barriers to optimal care; and suggestions as to how these barriers could be addressed. Forms were distributed to all women on admission to a postnatal ward over a two month period and completed prior to discharge. Of 500 eligible mothers, 151 (30.2%) responded to the anonymous open ended survey. A thematic analysis of comments revealed that women wanted specific information about mothering, the creation of a restful environment, adequate pain relief, practical assistance, education, and set visiting times. For new mothers, early discharge made the need for rest and information a high priority. Constraints within the public health care system and midwifery practice need to be examined to better serve mothers' needs. Midwifery practice within the context of early postpartum discharge should seek to better serve new mothers by giving high priority for rest and information requirements. PMID- 11887649 TI - Dealing with diversity: incorporating cultural sensitivity into professional midwifery practice. AB - In the Australian College of Midwives, Code of Ethics, Section 11. Practice of Midwifery, the following is stated "A. Midwives provide care for women and childbearing families with respect for cultural diversity while also working to eliminate harmful practices within those same cultures." However, it is difficult to know what is meant by "respect for cultural diversity". This paper presents the results of a critical review of the health literature. There is surprisingly little consensus about the meaning of terms such as cultural sensitivity and cultural appropriate care. Nor are there reflections on incorporating these concepts into practice. It could be argued that until there is greater clarity about these concepts and more discussion of how they may be used in practice, midwives would have to continue to rely on their individual knowledge and experience. PMID- 11887650 TI - The professionalisation of midwifery through education or politics? AB - The midwifery profession is redefining itself, with a national initiative to separate from nursing using several strategies at political, legal, educational and professional levels. These include lobbying for a Midwives' Act, a national approach to co-ordinate the education of student midwives, the introduction by the ACMI of competency based practice, the initiation of various models of practice and a three-year Bachelor of Midwifery. This paper argues that the educational strategy employed by midwifery is similar to that used by nursing. This strategy was overtaken by political and economic reforms within the health care sector. We argue that achieving professional dominance is not achieved simply through education but is fundamentally a political process. PMID- 11887651 TI - AMAP--the end of a beginning. PMID- 11887653 TI - Is surgical skill more important for clinical success than changes in implant hardware? PMID- 11887654 TI - Fixed mandibular restorations on three early-loaded regular platform Branemark implants. AB - BACKGROUND: Originally, the Branemark System was used as a two-stage surgical procedure. Comparable clinical results have made one-stage and early-loading concepts possible alternatives in the edentulous mandible. From the patient's point of view, the financial aspect of implant treatment is important. In an attempt to decrease financial burden, the reduction of surgical interventions and reduction of the number of implants could be considered. PURPOSE: This prospective multicenter study evaluated (1) the 1- and 3-year success rates of implants loaded within 1 month after one-stage surgery with a fixed 10- to 12 unit bridge on three regular platform Branemark System implants in the mandible, (2) the outcome of the prosthetic treatment, and (3) the opinion of patients regarding the treatment procedure. MATERIALS AND METHODS: Of 20 patients, 19 received five implants in the mandible, of which three were functionally loaded with the one-stage technique (group 1). The loaded implants were inserted in a tripodal position, one implant in the symphysis and two located anterior of the mental foramen in the bicuspid area. Two additional implants were inserted for safety reasons but were not intended to be included in the restoration. These two additional implants served as either an unloaded one-stage control implant (group 2) or an unloaded control implant installed with the submerged technique (group 3). Immediately after surgery, the implants were loaded with a relined denture. The patients received a 10- to 12-unit prosthetic reconstruction an average 31 days (range, 4-53 d) after surgery. Implant stability was clinically checked at 3, 12, and 36 months. Radiographs were taken at corresponding follow-up visits to calculate bone-to-implant level and marginal bone resorption. RESULTS: Six of 60 functionally loaded implants (10%) and 3 of 20 prostheses (15%) failed within the first year. The cumulative implant failure rate in group 1, both after 1 and after 3 years, was 9.5%. No implant failure occurred in the control groups 2 and 3. The average marginal bone level measured at 1 and 3 years was 1.6 mm (SD = 0.8 mm) and 2.1 mm (SD = 0.2 mm), respectively, for group 1; 1.5 mm (SD = 1.3 mm) and 2.4 mm (SD = 0.6 mm), respectively, for group 2; 0.8 mm (SD = 1.4 mm) and 0.7 mm (SD = 0.9 mm), respectively, for group 3. CONCLUSIONS: The results of treatment using three regular platform Branemark System implants supporting a fixed mandibular arch reconstruction were less favorable than the outcome that can be expected with a standard four- to six-implant with one-stage surgery. PMID- 11887655 TI - One-stage operative procedure using two different implant systems: a prospective study on implant overdentures in the edentulous mandible. AB - BACKGROUND: Evidence-based reports are needed to support the application of a one stage surgical protocol for unsplinted implants supporting mandibular overdentures. PURPOSE: To examine the feasibility and success of using two different dental implant systems (originally designed for two-stage operative technique) using a one-stage operative procedure in patients being rehabilitated with implant mandibular overdentures. MATERIALS AND METHODS: The study sample involved 24 edentulous subjects (aged 55-80 yr) randomly allocated to two different implant systems, one with a machined titanium implant surface (Steri Oss, Nobel Biocare, Goteborg, Sweden) and the other with a roughened titanium surface (Southern Implants, Ltd., Irene, South Africa). Two unsplinted implants to support implant overdentures were placed in the anterior mandible of all patients, using a standardized one-stage surgical and prosthodontic procedure. Primary stability and bicortical anchorage of the implants was mandatory before healing abutments were connected at the time of implant placement. Implant overdentures and their respective matrices were inserted following a standard 12 week healing period. Data relating to mobility tests, radiographs, and peri implant parameters were documented at 12, 16, and 52 weeks after surgery. RESULTS: A success rate of 95.8% for the Steri-Oss and 100% for the Southern Implants was found, without any statistically significant differences in the marginal bone loss. Significant changes in Periotest values were observed for both types between 12 and 52 weeks (p < .001). Minor changes were observed in the peri-implant parameters evaluated. CONCLUSIONS: These preliminary findings show a successful application of this one-stage approach for unsplinted implants supporting mandibular overdentures with Steri-Oss and Southern Implant Systems. PMID- 11887652 TI - What's new in defining hypertension and classifying hypertensive disorders in pregnancy. AB - Hypertensive disorders are the most common medical problems encountered by women during pregnancy and remain a major cause of maternal and fetal mortality and morbidity throughout the world. This article provides an update on the definition and classifications of hypertension in pregnancy and discusses the recent consensus statement from the Australasian Society into the Study of Hypertension in Pregnancy (ASSHP). A review of the relevance of the traditional triad of symptoms in the assessment of women for hypertensive disorders in pregnancy is also presented. An understanding of these issues will help midwives accurately assess women and ensure that appropriate management of pregnancies complicated by hypertensive disorders occurs. PMID- 11887656 TI - Histomorphometric and mechanical evaluation of the bone-tissue response to implants prepared with different orientation of surface topography. AB - BACKGROUND: Several studies have shown that in soft tissue, the orientation of surface topography affects cell response. However, the response of hard tissue to the orientation is not fully understood. PURPOSE: This study was undertaken to determine how orientation of the microstructure of implant surfaces influences bone healing. MATERIALS AND METHODS: Forty cylindrical implants were prepared with a dominant orientation of the microstructure (20 with a horizontal orientation and 20 with a vertical orientation) and investigated in vivo. Three methods for surface topographic characterization were used to investigate the topography in different resolution levels. RESULTS: Topographic analysis showed that a clear orientation was achieved at the implant surface, and a rougher surface structure was found on the vertically oriented grooves than on the horizontal ones. The implants were inserted in the tibia of 10 New Zealand White rabbits. Pull-out tests and histomorphometric analyses of ground-sections were made after 12 weeks of healing. The pull-out test showed a higher mean value for the vertically grooved implants, but the difference was not significant. Histomorphometric analyses showed no statistically significant differences between the horizontally and the vertically grooved implants when measuring bone to-implant contact or amount of bone area around the implant. CONCLUSIONS: No advantages could be found for either the horizontal or vertical orientation compared with each other, but further studies are needed in which implant roughness should be similar between the different topographies. PMID- 11887658 TI - Biologic and mechanical stability of single-tooth implants: 4- to 7-year follow up. AB - BACKGROUND: Single-tooth implants have been reported to achieve a high level of surgical and prosthetic success. However, close inspection of the literature reveals a paucity of data on the follow-up of single-tooth implants in function for 5 years or more. Since unsplinted implants may be considered to be subject to greater functional stresses, there is a need to report on the long-term biologic and mechanical integrity of such implant-supported restorations. PURPOSE: To report on the long-term follow-up of single-tooth implants, restored and in function for 4 to 7 years. MATERIALS AND METHODS: Twenty-seven Astra Tech single tooth implants were subject to a clinical audit to evaluate implant and prosthetic success as well as response of both hard and soft tissues over a 1- to 4-year follow-up. This current report presents data on the biologic and mechanical integrity of 23 of these single-tooth implants, which have been in function for up to 7 years (mean, 63 mo). Data are provided with respect to implant survival, maintenance of marginal bone levels, soft-tissue health, and the recording of any adverse events, including prosthetic complications. RESULTS: Only 14 implants in 13 patients were available for review, with no failures for this group of implants. One patient from the original group, who was lost to follow-up, was known to have suffered an implant failure. Furthermore, in accordance with established criteria, the remaining 13 implants that are not included in this report must at this time also be considered as potential failures. As such, the best-case scenario would be a 95.6% success rate for the 23 implants included in this review and the worst-case scenario would be a 60.8% success rate. Mean marginal bone loss measured 0.49 mm mesially and 0.76 mm distally, with a frequency of bone loss of 50%. Soft tissues were clinically healthy. There were few adverse events, with only one case of abutment screw loosening, detected at the 6-year review. In addition, crown decementation was recorded three times in two patients. CONCLUSIONS: It can be concluded that the Astra Tech single-tooth implant can achieve long-term biologic and mechanical stability when used to restore single missing teeth, over the long-term. PMID- 11887657 TI - Immediate occlusal loading of Branemark implants applied in various jawbone regions: a prospective, 1-year clinical study. AB - BACKGROUND: The original protocol for dental implant treatment ad modum Branemark was based on submerged healing prior to loading. For patients, immediate implant function could reduce cost and increase attractiveness of implant treatment. PURPOSE: The goal of this study was to evaluate the short-term success rate of immediately loaded implants placed in various regions of the jaws. MATERIALS AND METHODS: Forty-one patients received a total of 127 immediately loaded implants (76 maxillary and 51 mandibular). Seventy-one percent of the patients received their prosthetic restoration the same day and the others within 11 days. All prosthetic constructions were in full contact in centric occlusion. Clinical follow-up examinations were performed at 1 week, 2 weeks, and at 1, 2, 3, 6, and 12 months after implant loading. The study was completed 1 year after loading. RESULTS: Twenty-two implants were lost in 13 patients (including 7 maxillary implants lost in 1 patient). The cumulative success rate of the implants was 82.7% after 1 year of prosthetic loading. All sites with implant losses were re implanted, using a two-stage technique, with no further complications reported. Ninety-one percent of implants placed in regions other than the posterior maxilla were successful compared with 66% of implants placed in the posterior maxilla. Implants in patients with a parafunctional habit (bruxers) were lost more frequently than those placed in patients with no parafunction (41% vs. 12%). Implants subjected to guided bone regeneration were more successful compared with those not subjected to regeneration procedures (90% vs. 67%). CONCLUSIONS: The immediate loading concept is a realistic treatment alternative in various jawbone regions except for the posterior part of the maxilla. High occlusal loads should be considered a risk factor. On the other hand, implants in combination with bone defects frequently are penetrating cortical layers to a higher extent, thereby contributing to implant stability during the healing phase and consequently do not inevitably jeopardize the treatment result. However, further controlled clinical studies with larger sample sizes need to be performed to evaluate the influence of different parameters on treatment outcome. PMID- 11887659 TI - Application of oxygen ion implantation to titanium surfaces: effects on surface characteristics, corrosion resistance, and bone response. AB - BACKGROUND: The surface oxide layer of titanium plays a decisive role in determining biocompatibility. However, there are some reports demonstrating that the natural oxide film may not be sufficiently protective in the aggressive biologic environment. PURPOSE: The goal of this study was to examine the effectiveness of a thick oxide layer on corrosion resistance in vitro and the bone formation around titanium implants in vivo. MATERIALS AND METHODS: A plasma source ion implantation (PSII) method was used to increase the thickness of the surface oxide layer. Several instruments were employed to confirm the surface properties before and after the surface modification. Potentiodynamic polarization measurements in a phosphate-buffered saline (PBS) solution were carried out to investigate corrosion resistance in vitro. Bone formation around this surface-modified specimen was examined in a rabbit model and assessed in histomorphometry. RESULTS: Improved corrosion resistance was demonstrated by the potentiodynamic polarization measurements. Light microscopic histomorphometry showed that all implants were in contact with bone and had some proportion of bone within the threads at 4 weeks; however, there were no significant differences compared with as-machined controls. CONCLUSIONS: The results indicate that in spite of improved corrosion resistance in vitro, a thick oxide layer fabricated with the PSII method does not influence early bone formation around titanium implants in vivo. PMID- 11887660 TI - Who is treating them now? PMID- 11887661 TI - The delivery of orthodontic care in New Zealand. Part II: Analysis of a census of dentists. AB - Part I of this study reported the level and distribution of the supply of specialist orthodontic services in New Zealand. This paper focuses on the amount and variety of orthodontic services supplied by dentists. A questionnaire sent to all dentists in New Zealand sought information on the amount and type of orthodontic treatment carried out between 1 July 1998 and 30 June 1999. The reply rate was 80.9 percent. The majority of dentists carried out some form of orthodontic treatment, predominantly of a minor nature. A small number provided significant amounts of treatment, both simple and complex. The majority of orthodontic treatment and the majority of comprehensive fixed-appliance treatment were undertaken by orthodontists. One-quarter of all orthodontic patients in New Zealand were treated by dentists, irrespective of the complexity of treatment. Nearly a fifth of all full fixed upper and lower appliances, and nearly a third of all single-arch fixed appliances were placed by dentists during the study period. In general, male dentists, dentists over the age of 40, those who had attended an orthodontic continuing education course in the previous 5 years, and those who referred fewer patients to an orthodontist carried out more procedures, including those of a complex nature; they also had a higher average active orthodontic patient load. Wanting to be more or less busy had little influence on the amount or complexity of treatment performed. Dentists in regions with a low supply of specialist orthodontic services provided more comprehensive fixed appliance treatment and had a higher orthodontic patient load. However, the presence or absence of an orthodontist in an urban area seemed to have little impact on the complexity of treatment or the orthodontic patient load of dentists. Despite fewer orthodontists in secondary and minor urban areas, dentists in these areas did not have a higher orthodontic patient load, but carried out a wider range of procedures and more complex procedures than those in main urban areas. PMID- 11887662 TI - Quality of life and nutritional studies in Sjogren's syndrome patients with xerostomia. AB - People with xerostomia can experience significant difficulties eating some foods which, before the onset of the dry mouth, would have been easily consumed. The few studies in the literature indicate that such people become deficient in a variety of nutrients. In this study, supporting evidence was sought to confirm whether a New Zealand population of people with Sjogren's Syndrome and xerostomia was malnourished. Quality of life issues were measured in the same patients. There was no evidence of nutritional deficiency in the study group, nor did xerostomia seem to be important as a determinant of psychological distress or overall quality of life. As measured by the GHQ-12 score, xerostomic people without their natural teeth were, however, more psychologically distressed than those with a natural dentition. The importance of maintaining the natural dentition in xerostomia is emphasised by this latter result. PMID- 11887663 TI - Changed oral conditions, between 1963 and 1999, in the population of the Tokelau atolls of the South Pacific. AB - In 1999, an oral health survey was included in an assessment of the community oral health programme of the Tokelau Islands population. This provided a comparison with a similar survey in 1963. In a convenience sample of 386 children and adults, approximately 30 percent of the total population, the deciduous (number of df teeth) and permanent (number of DMF teeth) tooth scores across all age groupings were higher in 1999 compared with 1963. For 15- to 19-year-olds, the mean DMF scores were 8 and 1; and for 35- to 44-year-olds, the scores were 18 and 4 in 1999 and 1963 respectively. The prominent feature of the DMF scores for those over age 25 years was the numbers of missing (M) teeth. The mean number of M teeth at 20-24 years was 5 and 0, and at 35-44 years, 13 and 2 respectively in 1999 and 1963. Periodontal disease was endemic in adults in both surveys. A serious decline in oral health has occurred over the past 35 years. PMID- 11887664 TI - Informed consent for people with diminished capacity to consent. AB - This paper discusses the rights and responsibilities of patients and providers in New Zealand when a patient has diminished capacity to understand and provide consent for dental care. The Code of Health and Disability Services Consumers' Rights 1996, the Protection of Personal and Property Rights Act 1988, and the New Zealand Bill of Rights Act 1990 all affect decisions that relate to the provision of dental care for patients with diminished capacity to consent. A person's capacity to understand medical or dental procedures will vary with differing procedures and may vary at different points in time. Capacity to consent must be assessed by the dentist responsible for the proposed dental care in the first instance. Welfare guardians and power of attorney may exist for some patients with diminished capacity to consent, but the documentation of appointment to those roles should be read by the dentist to verify the extent of the appointment. When a patient is not competent to make an informed choice and give informed consent, treatment providers may provide care under Right 7.4 of the Code of Health and Disability Services Consumers' Rights 1996 if the appropriate provisions within the Code are satisfied. PMID- 11887665 TI - That little nudge. PMID- 11887666 TI - AIDS in New Zealand. PMID- 11887667 TI - The truth about dental insurance. PMID- 11887668 TI - The deal on digital: the status of radiographic imaging. AB - In the very near future, dentists will perform image acquisition, whether clinical or radiographic, using solid-state detectors or photostimulable phosphor plates. In addition, clinical still camera images, video camera images, and x-ray images will all be digital. At this time, medical imaging is almost completely digital and consumers are actively seeking digital products. Because information in a digital format can be stored, processed, and transmitted easily and efficiently by computers, traditional imaging receptors, such as conventional radiographic film, is slowly being replaced. This article will outline the various digital receptors and systems available for x-ray imaging, and discuss the impact of this paradigm shift on the dental professional. PMID- 11887669 TI - Recurring anterior facial swelling. AB - The nasolabial cyst was first reported in 1882. The most common clinical finding of the nasolabial cyst is that of an asymptomatic soft tissue swelling involving the canine fossa/nasal alar base region. The pathogenesis is controversial, but this cystic lesion most likely develops from the nasolacrimal duct. Microscopically, this cystic structure is composed of a fibrous capsule with an unremarkable layer of pseudostratified columnar epithelium. The treatment of choice is surgical excision, after which recurrence is rare. PMID- 11887670 TI - Nociceptive trigeminal inhibition--tension suppression system: a method of preventing migraine and tension headaches. AB - Dentists and physicians see countless patients suffering from various types of headaches. Various modes of therapy are used in an attempt to treat these patients. As a result of this study, it appears that a common factor to migraine and tension-type headaches may be chronic clenching. If this is the case, then dentists may treat headache patients more effectively than previously suspected through the use of a dental appliance. PMID- 11887671 TI - Surface disinfectants: read the labels. AB - There are a few basic long-standing guidelines, which can assist in the selection and safe use of chemical disinfectants (Table 2). Even though these were developed before the current availability of many of the premixed antimicrobial sprays and wipes, they remain applicable and most important, appropriate. PMID- 11887673 TI - Team esthetics: combining hard and soft tissue parameters to maximize esthetic results. PMID- 11887672 TI - Comparison of the effectiveness of several denture sanitizing systems: a clinical study. AB - The purpose of this clinical study was to test the effectiveness of three methods of decontamination on complete dentures. Dentures worn by patients for varying lengths of time were handled aseptically and treated with three different treatment modalities. The dentures were touched and sectioned and then retouched to a variety of microbiological media. The quantity of microbial growth was recorded and predominating microorganisms were identified using standard microbiological techniques. System A was found to consistently decontaminate and sanitize dentures worn by patients. System B and System C showed variable reduction of microorganisms. An unexpected spectrum of both pathogenic and opportunistic microorganisms was found in the dentures examined, including a wide range of gram-negative bacteria, gram-positive bacteria, and yeasts. A wide range of microorganisms must be considered when treating either oral or systemic diseases in denture wearers. Denture hygiene and decontamination are critical to the prevention of oral and systemic disease transmission. The dentures of ill patients must be considered as possible sources of pathogenic microorganisms. PMID- 11887674 TI - The MAS (mandibular advancement series) difference. AB - So now it may be seen what is meant by the term "The MAS Difference". The American orthodontic specialty has for the most part been slow, or even in isolated pockets, willfully resistant to expansion of orthodontic technique up out of the "Procrustean bed" of fixed appliance limitations to the panorama of attacking the teeth-bone-muscle triangle of malocclusion with separate techniques and appliance systems designed specifically for each. What makes this anathema to those of a broader view is that it is only with those expanded, combined fixed AND functional techniques, that one has any chance at all of rendering anything close to significantly successful treatment of major TMJ-pain-headache dysfunction chronic pain problems. And that is serious stuff. So, if you want to do "ortho" you had better know "TMJ". And, if you want to do "TMJ", at least TMJ to any meaningful degree, you had better know "ortho", and that means the discipline of functional orthodontics (or "maxillofacial orthopedics" if you will). The "why" of it is easily understandable once one truly understands the orthopedic (condylar displacement), myofunctional (Class II neuromuscular sling), and neurological (chronic repetitive compression nerve damage) aspects of the common functionally induced TMJ problem. The "how" of it all is another matter. That is why knowledge of a broad variety of various specific orthodontic techniques is required for the clinician because there are a vast variety of malocclusive situations with their own unique demands. But in the broadest sense, since a major portion of the orthodontic patient population suffers from somewhat of a skeletal Class II relationship, or "Class II effect" with respect to the joint, somewhere in the proposed treatment plan for these patients the clinician will have to consider some form of mandibular advancement series, whatever that series may be. But it is that MAS difference that sets that clinician and his or her specific treatment plan apart from the older, more restricted ways. It is a difference we must pay attention to, for Nature surely will. PMID- 11887675 TI - Refractive patients: TMJ splint therapy combined with open joint surgery: 60 patients; 117 joints. PMID- 11887676 TI - The conversion of vertical growth to horizontal. A prospective pilot study of twelve consecutive patients treated by two different methods. AB - Vertical growth may be a disadvantage for both facial appearance and long term dental stability. This paper reports a pilot study comparing the direction of facial growth following treatment by either fixed retractive appliances or by removable 'Orthotropic' appliances designed to reduce vertical growth. To meet ethical objections two different operators each treated a consecutive series of six patients with overjets exceeding ten millimeters. The resultant mean growth direction at Gnathion for the fixed retraction group was 67 degrees while in the Biobloc group it was 46 degrees. PMID- 11887677 TI - The Garcia Distraction Appliance: treatment of the TMD patient with an anterior open bite. AB - A temporomandibular joint case with an anterior open bite and a high mandibular plane angle was treated by erupting the posterior teeth after creating a posterior open bite using a Garcia Distraction Appliance and a Garcia Pivot Splint. PMID- 11887678 TI - Life change events, hope, and self-care agency in inner-city adolescents. AB - PROBLEM: Little is known about the relationships among life change events, hope, and self-care agency in inner-city adolescents METHODS: Adolescents (N = 202) attending two inner-city high schools in Miami (M = 15.9 years; 85% African Americans; 55% females). Measures used included the Adolescent Life Change Events Questionnaire, Miller Hope Scale, and Denyes Self-Care Agency Instrument. FINDINGS: A significant positive correlation was found between hope and self-care agency. No correlations were found between life change events and hope or life change events and self-care agency. Hope accounted for 20.8% of the variability in self-care agency. CONCLUSIONS: Hope is a significant factor in self-care agency and may be a coping strategy for inner-city adolescents who experience multiple life change events. PMID- 11887679 TI - The importance of fulfilling unmet needs of rural and urban adolescents with substance abuse. AB - PROBLEM: Because of the negative impact on health of drug and alcohol use, this study examined adolescent needs and substance use. METHODS: Subjects (N = 191) were adolescents aged 14 to 19 in a rural and an urban high school. A modified version of the Need Subscale from the Addiction Research Center Maturation Scale measured a feeling of satisfaction related to meeting basic needs, and an investigator's prepared questionnaire elicited current use of alcohol, nicotine, and marijuana. FINDINGS: Individuals with feelings of unmet needs were more likely to be current drinkers. Rural/urban residence was not a significant predictor in a multivariate analysis, but religiosity was. CONCLUSIONS: A feeling of unmet needs seems to be an important factor in adolescent substance use. PMID- 11887680 TI - Attention deficit hyperactivity disorder: national resources. PMID- 11887681 TI - Helping children understand death. PMID- 11887682 TI - "Growing up" children: current child-rearing practices among immigrant Jamaican families. AB - PURPOSE: To explore and describe current child-rearing practices within immigrant Jamaican families living in the United States. METHODS: This ethnographic study (N = 16, ages 9-60+) sought to gain an emic perspective of participants' experience with the phenomenon. Interpretive interactionism as a method looked at the process of child rearing in order to uncover meaning of significant events related to this practice. FINDINGS: Historical factors affecting the parents' own growing up experience and their experience with and perception of dominant culture values guided their child-rearing practices. CONCLUSIONS: The goal of child rearing for this immigrant group centered on raising a spiritual, respectful, knowledgeable, and competent child who would be equipped with necessary skills to survive in this society. PMID- 11887683 TI - [Affirming our knowledge: the stakes and the conditions of success]. PMID- 11887684 TI - [Hospitality as a philosophy of caring]. PMID- 11887685 TI - [Research: privileged voices in the development of nursing knowledge]. PMID- 11887686 TI - [Society and science: choices and stakes]. PMID- 11887689 TI - A memorable convention, and a general meeting facing the future with solidarity. PMID- 11887688 TI - [Supervision of undergraduates at the formative stage or of nursing students at orientation]. PMID- 11887690 TI - Reaffirming a primary commitment to nonmaleficence. PMID- 11887692 TI - How I survived outplacement and lived to tell about it. Outplacement survival tips. PMID- 11887691 TI - The legal forum. PMID- 11887694 TI - When surrogate decision-making is not straightforward. Guidelines for nurse administrators. AB - When ethical or pragmatic questions arise concerning surrogate decision-making, nurse administrators often are consulted, so they must be knowledgeable about current legislation as well as the surrogate decision-maker's role and ethical obligations. Case exemplars are presented in this article to facilitate the reader's awareness on how value conflicts and communication failures can complicate surrogate decision-making. Recommendations for clinical practice and research are presented. PMID- 11887696 TI - Space analysis in fixed prosthodontics--normal parameters. AB - Partial edentulism is a common clinical situation encountered by dentists in their daily practice. If left untreated, the early loss of molars or any other tooth often leads to migration of the neighboring teeth, leaving abnormal edentulous spaces to be restored. In this article, guidelines are described for analyzing simple and complex edentulous spaces as well as how to determine the size of pontics to be used through a simple sequence of steps. Within the context of this article, clinical examples are shown that illustrate the simple, normal parameters of this technique, which is also applicable to more complex restorative situations. PMID- 11887687 TI - [The challenges of practical nursing in the 21st century]. PMID- 11887695 TI - Managed care and the evolution of patient rights. AB - Managed care has grown into the driving force for coordinating and managing healthcare delivery and its financing. At the same time, major initiatives have gained momentum in societal and legislative efforts regarding patient rights. This article provides a historical, legal, and philosophical review of the major events that have shaped the evolution of healthcare services and development of the patient rights issue. As the healthcare industry finds itself in a state of needed reform, the future evolution of managed care in light of the patient rights issue will have important implications for legislators, clinicians, and healthcare organizations as they struggle to provide quality care and improve cost controls. PMID- 11887697 TI - Maxillary canine impactions: orthodontic and surgical management. AB - With the exception of the maxillary and mandibular third molars, the maxillary canine is the most commonly encountered impaction. In North America, palatally located impactions appear to predominate over labially impacted canines and occur more frequently in women. Because of the common nature of this phenomenon, the general dentist should know how to properly diagnose and manage potential disturbances in the eruption of the maxillary canine. If diagnosed early, and when clinically and radiographically indicated, extraction of the deciduous canine may help correct the eruption path of the permanent tooth and prevent impaction. This article reviews the etiology, prevention, and surgical and orthodontic management of impacted cuspids. Because the general dentist is often the first dental care provider to come in contact with patients with impacted canines, knowledge about this common dental anomaly is essential to provide proper comprehensive therapy. PMID- 11887698 TI - Clinical advantages of tapered root form dental implants. PMID- 11887699 TI - Advances in abrasive technology--prophylaxis pastes. AB - A newly developed prophylaxis paste that contains perlite as an abrasive medium was compared to conventional prophylaxis pastes with regard to relative dentin and enamel abrasion, cleaning ability, and polishing power. Rubber cups and nylon brushes were used as paste carriers. Water, flour of pumice, and a dentin polishing paste served as controls. The Prophylaxis Paste Index was created to assess the clinical potential of the various prophylaxis pastes more accurately. The index was computed for dentin as cleaning ability divided by relative dentin abrasion x surface roughness (in Ra) x 10, and for enamel as cleaning ability divided by relative enamel abrasion x surface abrasion (in Ra). On dentin and enamel, the perlite-based prophylaxis paste consistently yielded low relative dentin and enamel abrasion values, a good cleaning ability, and low surface roughness scores with both rubber cups and nylon brushes. Of all tested prophylaxis pastes, the perlite-based prophylaxis paste excelled in the Prophylaxis Paste Index on dentin and enamel and with both rubber cup and nylon brush applications. Therefore, a perlite-containing prophylaxis paste may have broad professional therapeutic indications and may be used universally as a single paste to clean and polish both dentin and enamel. PMID- 11887701 TI - Breast feeding practices among female health functionaries. AB - Fifty-nine married Female Health Functionaries, which included forty-eight ANMs and eleven LHVs working in District Kangra of H. P. have been interviewed through self-administered questionnaire during August 1999. Breast Feeding (BF) practice is universal. Average duration of initiation BF after delivery is six hours. It is continued for 23 months on an average. Practice of prelacteal feed and bottle feeding is prevalent. In 17% of respondents difficulty has been experienced in initiating breast-feeding. PMID- 11887700 TI - Occupational health nurse: an overview. PMID- 11887702 TI - Nursing the elderly--do the needs of the elderly differ? PMID- 11887704 TI - [The first examination at the obstetric department--the most important infant physician contact? A chance of early detection of curable disorders]. PMID- 11887705 TI - [The HSAN's judgments are mostly fair and judicious. But human errors do occur]. PMID- 11887706 TI - [Screening for congenital cataract is best done at the obstetrical department. Complementary examination at the child health center is recommended]. AB - 72 children born between January 1992 and December 1998, diagnosed with congenital cataract (CC) and operated on before the age of one, were included in a retrospective study. Three regions with different screening strategies were compared: one region with screening at the maternity wards, one at well baby centres and one without a screening programme. Eye screening at the maternity wards resulted in a significantly lower mean age at detection and operation compared to the other two screening strategies. Eye examinations should also be made standard procedure at well baby centres, in order to minimize the number of undetected cataracts. PMID- 11887707 TI - [Crisis intervention after emergency Cesarean section can result in a more positive experience of the labor]. PMID- 11887708 TI - [Swedish study of home accidents: children with young mothers and children in families on welfare are at greater risk]. AB - In this study we used data from Swedish national registers to describe socio demographic patterns of hospital admissions after injuries in the home (poisonings, falls, scald injuries and ingestion/intrusion of foreign objects) in children 0-2 years of age. Infants and toddlers in families with young mothers and in families on social welfare were at particular risk, while children with foreign-born parents and children in single parent households were at greater risk for scald injuries only. The relative risk for poisonings, scald injuries and ingestion of foreign objects respectively was found to be correlated to quite specific ages, a finding which should be used in timing of parent counseling. PMID- 11887709 TI - [How competently are HSAN errands proceeded?]. PMID- 11887703 TI - AIDS and reality. PMID- 11887710 TI - [Incidence of cardiovascular diseases can be influenced by factors experienced during early life stages]. PMID- 11887711 TI - [Cost-effectiveness of bisoprolol in chronic heart failure]. AB - The cost-effectiveness of adding the beta blocker bisoprolol to standard treatment in patients with congestive heart failure was investigated, based on data from the Cardiac Insufficiency Bisoprolol Study II (CIBIS II). The medical resource consumption from CIBIS II was combined with Swedish cost data for medication and hospitalisations. Costs of added years of life, i.e. consumption net of production, were also included in the analysis. The health effects were measured in terms of gained years of life. The results of the analysis show that the cost-effectiveness of bisoprolol compares favourably with that of other cardiovascular treatments. Without the inclusion of costs of added years of life, the cost-effectiveness was in the range of SEK 3,351-13,096 per gained year of life, and with the costs of added years of life included, the cost-effectiveness was in the range of SEK 137,533-147,278 per gained year of life. PMID- 11887712 TI - [Good opportunities to prevent contact eczema through legislation]. AB - Allergic contact dermatitis is often caused by allergenic substances in products. Legislation can be a useful tool in the prevention of contact dermatitis, by limiting skin exposure to contact allergens in chemical products, cosmetics and hygiene products and nickel-containing objects in contact with the skin. A brief review is given of the most important EU Directives, some national regulations and standardisation projects aiming at the prevention of contact dermatitis. It is essential that medical scientific experts support, with their expert knowledge, national and European authorities. PMID- 11887713 TI - [Survey of personal injuries caused by dogs and cats in Umea. Walking the dog was the activity most often related to injury]. AB - Injuries due to interaction with dogs and cats are not uncommon, and constituted 1.4% of all injuries in an unselected policlinical and inpatient material. This material was collected from a population of 135,000 people in Northern Sweden, and during 2 years 280 patients with injuries related to these domestic animals were found (Ndogs = 212, Ncats = 68). The injuries were in most cases caused by bites and by falls. The former were more common among men, the latter more common among women and related to fractures and dislocations. In contrast to many previous investigations, bite injuries were more common among adults than among children, possibly related to national differences in the keeping of dogs. Most dog related injuries were caused by the owner's dog. Walking the dog was the activity most often related to personal injury. The cost for inpatient cases (treated at the University Hospital, Umea) caused by dogs and cats is of the same magnitude as the cost due to motorcycling in traffic, but less than the cost for injuries sustained during horseback riding and other horse related activities, for soccer, snowmobiling and for assaults. PMID- 11887715 TI - [When politicians don't dare or don't want--we must for the patients' best]. PMID- 11887718 TI - [No one should suffer because of "cellular changes"]. PMID- 11887716 TI - [Taking care of the budget or human beings?]. PMID- 11887717 TI - [Central stimulants in the treatment of children with ADHD: unknown long-term effect and uncertain pharmacological documentation]. PMID- 11887719 TI - [Occupational environment and risk assessment for women employed in shipping industry]. PMID- 11887714 TI - [A Sida-supported project for improved reproductive health in southern Africa. The Samousa-project honours the principle of mutuality]. PMID- 11887721 TI - [A professional group helps district practitioners to cope with emotional problems]. PMID- 11887720 TI - [Computers and amalgam are the most common causes of hypersensitivity to electricity according to the sufferers' reports]. PMID- 11887722 TI - [Don't let psychiatry to be caught in its own trap]. PMID- 11887723 TI - [Sports physicians fight against sudden cardiac death of athletes. To drop dead is not allowed]. PMID- 11887724 TI - [What is better: virtual or endoscopic coloscopy? The colonoscope has not become superannuated so fast!]. PMID- 11887726 TI - [Fatty liver and raised gamma-GT: it can happen to abstainers, too]. PMID- 11887725 TI - [Noninvasive look into the heart. For us it is a fulfilled dream]. PMID- 11887727 TI - [Grave deficiencies in the diagnosis and therapy of osteoporosis. Expensive fracture landing]. PMID- 11887728 TI - [Disputed new thinking in osteoporosis. Bone corrosion by sausage and cheese?]. PMID- 11887730 TI - [Borderline disorders in childhood and adolescence]. AB - Borderline disorders are situated in the area between neurosis and (schizophrenic) psychosis. In psycho-analytical terms, a "borderline personality organization" is characterized by the following criteria: weak sense of identity, poor reality control, permeable ego boundaries, multifarious defense reactions to impulsivity, disturbed object-relatedness with inappropriate relationship to the environment, and the assumption of a disorder/trauma during early childhood. Some 90% of the children have a history of traumatization in the past, e.g. a severe illness or sexual abuse. Various models are used to explain the multifarious disorders. What can be compensated in childhood via so-called "emergency maturation" and adaptation to the prevailing situation, is far more difficult from early adolescence onward. Then, external regulators, such as alcohol or drugs play a greater role in the re-establishment of inner equilibrium. PMID- 11887729 TI - [Disturbed early bonding and its consequences]. AB - During the course of its first year of life, the infant develops a strong attachment to the person with whom it has most contact (usually the mother), to whom it turns for consolation when in pain, or protection when threatened. If the infant's bonding needs are not adequately met, rage, disappointment, or ambivalent feelings towards that person may result. The trauma of separation or loss, pronounced emotional deprivation, and abusive experiences are a major reason for disordered bonding. Early diagnosis, psychotherapeutic treatment of children with attachment problems and parent counseling or treatment are essential to prevent the problem from becoming chronic and to promote the development of adaptive bonding patterns. PMID- 11887731 TI - [Substance addiction in children and adolescents]. AB - Youngsters in Germany are becoming addicted at an ever younger age--not only to alcohol and tobacco, but also to illegal drugs. The underlying causes are manifold: in the first place peer duress, curiosity and a search for solace render the child susceptible to the promise of the "panacea". It is of importance to distinguish between "dependency syndrome" and "harmful use". The diagnosis of substance dependence is based on typical clinical features and on detection of the substance in the blood or urine. If dependence is present, the first step is usually detoxification, with efforts undertaken to initiate a competent pediatric/adolescent psychiatric diagnostic work-up and treatment as early as possible. The leading preventive measure is to provide relevant instruction on the dangers of the legal drugs alcohol and tobacco. PMID- 11887732 TI - [Emergency blood cough. To stop the bleeding: to detect the cause and localization]. PMID- 11887733 TI - [When blood pressure does not lower at night. Stroke risk is greatly elevated]. PMID- 11887734 TI - [What, how much and for whom? Movement by prescription]. PMID- 11887736 TI - [Therapy of erectile dysfunction. New, highly selective PDE-5 inhibitor]. PMID- 11887735 TI - [Erectile dysfunction. New therapy options ante portas]. PMID- 11887737 TI - [Insurance is pressured now with disease management. KBV warns: false start breaks the solidarity community]. PMID- 11887738 TI - [Disease management program. Nobody must join but nobody can ignore it]. PMID- 11887739 TI - [Chronic bronchitis. What antibiotic in acute exacerbations. Good and fast effectiveness is needed]. PMID- 11887741 TI - [Regardless of therapy only every fifth person has normal blood pressure values. Morning hours are deadly]. PMID- 11887740 TI - [Effective not only in rheumatism and arthrosis. Rofecoxib fights also acute pain]. PMID- 11887743 TI - [Blood pressure self-determination. How can false values be prevented?]. PMID- 11887746 TI - [Self care for pregnant women. pH-determination prevents premature labor]. PMID- 11887744 TI - [Migraine has to be treated. Otherwise a stroke threatens]. PMID- 11887745 TI - [Smoking, drinking and mental laziness. It hastens cognitive cut-down]. PMID- 11887751 TI - Factors influencing the conversion of carotenoids to retinol: bioavailability to bioconversion to bioefficacy. AB - Vitamin A (retinol) is a necessary nutrient for vision, reproduction, growth, and immune function. Pro-vitamin A carotenoids are an important source, especially in developing countries. While preformed vitamin A is readily available from foods, carotenoids are much more difficult to assimilate. A number of factors have been identified that either enhance or hinder the bioavailability of carotenoids. These have been presented in the literature and given the mnemonic SLAMENGHI by some researchers. The following factors are summarized: Species of carotenoid, molecular Linkage, Amount in the meal, Matrix Properties, Effectors, Nutrient status, Genetics, Host specificity, and Interactions between factors. Identifying which of these are key issues for the general public, and promoting the increased consumption of fruits and vegetables with moderate and high levels of pro-vitamin A carotenoids, are important to vitamin A status and overall good health. PMID- 11887747 TI - Calcium bioavailability in relation to bone health. AB - A well established stable isotope technique exists for measuring calcium absorption from single foods and meals, but the long term effects of calcium on bone health cannot be assessed from acute bioavailability studies. Bone health depends primarily on the degree of mineralization, measured as bone mineral density (BMD), and phenotypic variations depend on genetic and environmental factors including calcium supply. Since almost all retained calcium is used for bone mineralization and remodeling, BMD can be used as a long-term (> six months) marker of dietary calcium bioavailability. However, BMD is a very insensitive marker of calcium bioavailability, so its use in dietary intervention studies is restricted to periods of significant bone growth or loss. Biochemical markers of bone metabolism may be used to predict the overall bioavailability of dietary calcium over a shorter time period (> four weeks), but they have a high coefficient of variation, so may not be appropriate for some dietary intervention studies. A group of European laboratories is currently developing an alternative approach using a long-lived radioisotope (41Ca) to label bone calcium and to directly measure the rate of calcium loss from urinary excretion data. The efficiency of calcium absorption is inversely related to intake; whole body balance of the mineral is dependent on rates of absorption and excretion and limited by calcium-binding substances in the gut. Dietary data and indirect measures of bone health indicate that bioavailability is important when habitual intakes are low, especially during periods of bone growth or loss. Further research is required to quantify the effects of major dietary modulators of calcium balance on bone health and to understand their relationship with genetic and physiological variables. PMID- 11887742 TI - [The third-generation antihistaminics. Can they prevent the tier change?]. PMID- 11887753 TI - Bioavailability--a time for reflection. PMID- 11887748 TI - Iron status influences the efficacy of iodine prophylaxis in goitrous children in Cote d'Ivoire. AB - In the developing countries of Africa, many children are at high risk for both goiter and iron-deficiency anemia (IDA). Because iron (Fe) deficiency can have adverse effects on thyroid metabolism, Fe deficiency may influence response to supplemental iodine in areas of endemic goiter. Therefore, our aims were to determine: 1) if goitrous children also suffering from IDA could respond to oral iodine supplementation; and 2) if Fe supplementation in goitrous children with IDA would improve their response to oral iodized oil and iodized salt. First, we compared the efficacy of oral iodized oil in two groups of goitrous children: a nonanaemic group vs. an IDA group. The therapeutic response to iodized oil was impaired in the goitrous children with IDA. Second, an open trial of Fe treatment in goitrous children with IDA improved their response to oral iodized oil. Finally, in a randomized double-blind trial, goitrous, Fe-deficient children consuming iodized salt were given Fe supplementation or placebo. Fe supplementation improved the efficacy of the iodized salt. In these studies, both anatomic (thyroid size) and biochemical (TSH, T4) measures indicated that iodine significantly improved thyroid function in the nonanaemic children compared to the Fe deficient children. Iodine was less efficacious in children with lower Hb at baseline and in those with a poorer response to Fe. The data suggest that a high prevalence of IDA among children in areas of endemic goiter may reduce the effectiveness of iodine prophylaxis. PMID- 11887749 TI - Glucosinolates: bioavailability and importance to health. AB - Epidemiological studies suggest that brassica vegetables are protective against cancers of the lungs and alimentary tract. Cruciferous vegetables are the dietary source of glucosinolates, a large group of sulfur-containing glucosides. These compounds remain intact unless brought into contact with the enzyme myrosinase by pests, food processing, or chewing. Myrosinase releases glucose and breakdown products, including isothiocyanates. These highly reactive compounds are potent inducers of Phase II enzymes in vitro. Isothiocyanates also inhibit mitosis and stimulate apoptosis in human tumor cells, in vitro and in vivo. To understand and exploit such effects it is important to determine the routes of absorption of glucosinolate breakdown products, their metabolism, and delivery to systemic tissues. Glucosinolates can be gained or lost by vegetables during storage. They may be degraded or leached during processing, or preserved by thermal inactivation of myrosinase. Glucosinolates are broken down by plant myrosinase in the small intestine or by bacterial myrosinase in the colon. Isothiocyanates are absorbed from the small bowel and colon, and metabolites are detectable in human urine two to three hours after consumption of brassica vegetables. Interpretation of epidemiological data and exploitation of brassica vegetables for human health requires an understanding of glucosinolate chemistry and metabolism, across the whole food chain, from production and processing to the consumer. PMID- 11887756 TI - Strange days indeed. PMID- 11887755 TI - Biotechnology: a solution for improving nutrient bioavailability. AB - Biotechnology strategies are now available to improve the amount and availability of nutrients in plant crops. Those strategies include simple plant selection for varieties with high nutrient density in the seeds, cross-breeding for incorporating a desired trait within a plant, and genetic engineering to manipulate the nutrient content of the plant. In plant cross-breeding, all genes of the parent plants are combined and the progeny have both desirable and undesirable traits. To eliminate undesirable traits, plant breeders "back-cross" the new plant varieties with other plants over several generations. This technique, called hybridization, has been used to create varieties of low-phytate corn, barley, and rice. Using the techniques of genetic engineering, the gene(s) encoding for a desired trait(s) in a plant are introduced in a precise and controlled manner within a relatively short period of time. Golden rice, containing carotenoids, and rice with higher amounts of iron, are two examples of genetically engineered plants for improved nutrition. Genetic engineering has tremendous potential for revolutionizing nutrition. However, public concerns regarding safety, appearance, and ethics must be overcome before these products can be effectively introduced into the food supply. PMID- 11887754 TI - Influence of lifestyle on vitamin bioavailability. AB - In this review the effects of lifestyle factors, especially alcohol consumption, on vitamin bioavailability are summarized and discussed. Alcohol effects are clearly dose-dependent. Excessive chronic alcohol intake is generally associated with vitamin deficiency (especially folate, thiamine, and vitamin B6) due to malnutrition, malabsorption, and ethanol toxicity. Effects of moderate alcohol use are mainly explained by a lower vitamin intake. In the case of vitamin A and beta-carotene, effects on post-absorptive (lipoprotein) metabolism have been demonstrated. In one diet-controlled crossover study, alcohol consumption resulted in an increase in the plasma vitamin B6 (PLP) content, especially after beer consumption (containing vitamin B6), but also after wine and spirit consumption (not containing vitamin B6). Smoking is also associated with a lower dietary vitamin intake. In the case of vitamin C, B12, folate, and beta-carotene, evidence has been presented for effects on postabsorptive metabolism, due to smoke-induced oxidative stress and/or vitamin inactivation. For vitamin E a direct effect of smoking on absorption has been demonstrated. There is no convincing evidence that low-fat diets negatively affect fat-soluble vitamin absorption, but cholesterol-lowering compounds (diets), or unabsorbable fat substitutes, may do so. Vitamin bioavailability may be compromised from certain vegetables (particularly raw), and/or from high-fiber foods, because of limited digestion and inefficient release of vitamins from the food matrix. PMID- 11887759 TI - The Healthwatch Award speech: what patients are for. PMID- 11887760 TI - The plain lawyer's guide to bioterrorism. PMID- 11887758 TI - Animals and the law. PMID- 11887750 TI - A healthy diet rich in carotenoids is effective in maintaining normal blood carotenoid levels during the daily use of plant sterol-enriched spreads. AB - Blood cholesterol levels are affected by diet and in particular by the type and amount of fat intake. In recent years, vegetable oil spreads containing plant sterols/stanols (as their fatty acid esters) have been developed. Numerous clinical trials on spreads with added plant sterols/stanols have shown that they have much greater cholesterol-lowering properties than conventional vegetable oil spreads. Plant sterols decrease both dietary and biliary cholesterol absorption in the small intestine, with a consequential increase in excretion of cholesterol. It is also recognized that plant sterol/stanol-enriched, cholesterol lowering spreads, if consumed regularly, may induce a 10-20% decrease in plasma carotenoids, adjusted for changes in plasma lipids. A 10-20% decrease in plasma carotenoids falls well within the seasonal variation observed in individuals. Our current understanding of the physiological functions of carotenoids does not indicate any health risk associated with the slight decrease in their blood levels due to the intake of plant sterol/stanol. The questions that have been raised, though, are how plant sterols/stanols affect plasma carotenoid levels, and in addition, what quantity of fruits and vegetables (the richest dietary sources of carotenoids) would have to be consumed to improve plasma carotenoid levels? The current mini-review covers the cholesterol-lowering effect of plant sterols, their mechanisms of action and effect on blood carotenoids, and concludes with the potential heath benefits of daily intake of plant sterol enriched spreads. PMID- 11887762 TI - [Asthma in Africa: guidelines must be applied]. PMID- 11887764 TI - [Tuberculosis in the world: status and perspective in the year 2001]. PMID- 11887761 TI - [The pollen we inhale...]. PMID- 11887763 TI - [A gene for primary pulmonary artery hypertension]. PMID- 11887765 TI - [Flu vaccine and anti-neuraminidases. Synergy or concurrence?]. PMID- 11887766 TI - [Indications and application of exercise tests in children]. AB - Exercise tests are routinely used in children to assess cardio-respiratory and muscular adaptations to exercise. However these tests are of relatively recent use, and there is a lack of standardization and of relevant data in large groups in this population. The aim of this paper was to specify the common medical indications of exercise tests in children, to propose standardized protocols of these tests in some of the most common pathological situations as: exercise induced asthma, chronic respiratory diseases (bronchopulmonary dysplasia, cystic fibrosis), muscular diseases. These tests can provide clinically relevant parameters only when they are used in strict conditions of standardization. PMID- 11887767 TI - [Maximal oxygen uptake in healthy children: factors of variation and available standards]. AB - Aerobic physical fitness, in children, is assessed by measurement of the maximal oxygen consumption during exercise testing. Representative norms of the studied population are required for interpretation. The aim of this article is to specify and review the available VO2max norms and factors of variation, including: sex, anthropometric characteristics (height, lean body mass and weight) and physical activity level. Ideally, VO2max norms should include lean body mass and physical activity with an allometric equation. Since such norms do not exist today, interpretation remains difficult. In France, the must satisfactory norms for non trained children include body mass without an allometric equation (boys: 47 +/- 2 ml.mn.-1 kg-1, girls: 40 +/- 3 ml.mn.-1 kg-1 with a post puberty decrease). Further studies on VO2max norms that include lean body mass and a physical activity questionnaire are required to improve exercise test interpretation in children. PMID- 11887757 TI - The clinical disputes forum. PMID- 11887752 TI - Folate bioavailability and health. AB - The vitamin folate has been largely responsible for a fundamental shift in our perception of the role of vitamins in maintaining health. Ten years ago, two independent clinical trials showed that supplementing a woman's diet with folic acid before and during early pregnancy reduced the prevalence of neural tube defects (NTDs) by more than 70%. A remarkable aspect was that folic acid supplementation was not correcting a clinical deficiency in most of these women. It was later shown that the risk of having an NTD-affected birth was negatively associated with maternal red cell folate status, and the level of risk varied throughout the normal range, suggesting an interaction of genetic factors with folate nutritional status. It now appears that an insufficient folate status might contribute to risk of developing a variety of medical conditions throughout an individual's lifetime, from certain congenital malformations and poor pregnancy outcomes to cardiovascular disease, some malignancies, and neurological dysfunction of the elderly. Thus, an alternative view of folate nutrition has emerged. This view goes beyond the idea of a dietary requirement to prevent signs and symptoms of clinical deficiency towards one of achieving an optimal status to reduce risk of certain chronic diseases, and includes the concept that an individual's genetic make-up may substantially affect the vitamin status that they can achieve without supplements or fortification. PMID- 11887768 TI - [Evaluation of respiratory intensive care units at pneumology services]. AB - Audits should be conducted in respiratory intensive care units (ICU) as in all other ICU using patient-specific indicators to assess medical activity and quality of care. However, other criteria, such as those developed by the SRLF ("Societe de Reanimation de Langue Francaise"), should also be used. These criteria include the description of the patients previous health status, prognosis of underlying diseases, the SAPS I or SAPS II severity score at admission, the omega or TISS therapeutic scores, and the PRN index of health care burden. Medial and administrative audits are conducted using diagnosis-related groups (DRG) and case mix classification. The DRGs are used to establish an aggregate index of activity (ISA points) which contribute to the complex mechanism of budget allowance. Unfortunately, the French DRG case mix system does not provide an appropriate description of the costs of ICU stays. One of the special features of respiratory ICUs is related to patient flow. Patients are referred from a respiratory unit and discharged to a respiratory unit or a respiratory rehabilitation center. Readmissions are frequent. Many patients are also admitted only for diagnosis or a respiratory procedure requiring close surveillance. The SRLF criteria, which take into consideration all of these features, should always be used for the evaluation of quality of care. The French Society of Lung Disease (SPLF) has proposed specific standards of quality for respiratory ICU. We present here examples issuing for the ICU of the Hotel-Dieu Hospital in Paris. The results show that non-specific national indicators, in combination with other indicators specific for respiratory ICU, provide appropriate audit data. PMID- 11887769 TI - [Pollinosis and use of anti-allergic drugs: ragweed in the Rhone-Alpine region]. AB - Ragweed pollinosis was first described in the middle Rhone valley in the early sixties. Prevalence has increased steadily since that time. The aim of the present study was to determine whether this phenomenon is perceptible in terms of anti-allergic drug sales (eye drops, nasal spray, oral antihistamines) and if so, to compare sales with those related to hay fever. For this purpose, monthly sales in July, August and September were compared between an area of high infestation, a control area and national data for France. The comparisons showed a steep rise in sales for all kinds of anti-allergic drugs related to ragweed pollens, the level being comparable with that observed for gaminaceae. Surprisingly we were unable to find this type of data in the literature despite the evident usefulness of an epidemiological and economical analysis of the progression of pollen allergy. PMID- 11887771 TI - [Evaluation of management of asthma African adults. National survey among general physicians from Ivory Coast]. AB - A cross sectional study of asthma management of adults in Africa based on an anonymous questionnaire was conducted among 149 general practitioners working in the principal 18 cities in the Ivory Coast located in 6 geographical regions drawn for the purpose of this study. Data collected included: complementary tests prescribed at first diagnosis, typical prescriptions for acute asthma and for long-term control, knowledge of metered-dose inhalation techniques, regular measurement of FEV1 and prescription of sports activities. Forty-one percent of the physicians were unaware of the usefulness of expiration volume measurements and FEV1 measurements were prescribed by only 8%. The most widely prescribed tests were chest x-ray, blood cell counts, and ENT consultation. Lung function tests were requested by 12.75% of the physicians and skin allergy tests by 10.74%. Beta 2 agonists were prescribed for acute asthma, basically by inhalation or subcutaneous administration and were generally combined with corticosteroid therapy. Nebulization was rarely prescribed. Aberrant prescriptions (delayed release tablets, corticosteroid sprays, delayed release corticoidsteriods) were also given. For long term management, delay release beta 2-agonists were most widely used. Delayed release corticosteroid injections were prescribed by 18.5% of the physicians and metered dosers by 14%. Dry powders were rarely used. These findings emphasize the poor management of asthma in Africa. Directives taking into account the social and economic setting in the Ivory Coast should be initiated. PMID- 11887772 TI - [Spontaneous rupture of the esophagus or Boerhaave syndrome. Report of 3 cases and review of the literature]. AB - A triad of signs constitutes Boerhaave syndrome: forceful vomiting, chest pain and subcutaneous emphysema. The syndrome results from spontaneous rupture of the oesophageal wall leading to an oeso-pleural or oeso-mediastinal fistula. Positive diagnosis is established with a water-soluble swallow, sometimes coupled with computed tomography of the thorax. Boerhaave syndrome is a surgical emergency. We report three cases of spontaneous rupture of the oesophagus and analyze the importance of emergency surgery as well as emergency treatment of the sepsis, an important prognosis factor. PMID- 11887770 TI - [Skin sensitization to pollens in Morocco. Multicenter study]. AB - The objective of this study was to determine the prevalence of skin sensitization to certain pollens in some regions of Morocco. This multicentric study involving 10 centres included a consecutive series of 640 patients consulting for the first time between february and april 1998 for asthma and/or rhinitis and/or conjunctivitis. Skin prick tests (SPT) were performed with stallerpointes (Stallergenes) and interpreted according to standard procedures. Pollinosis was ascertained when SPT was positive to one or more pollens. Epi-Info was used to analyze the data. SPT were positive in 75.9% and prevalence of sensitization to pollens was equal to 28% (179 cases). It ranked second after house dust mites (63%), olive, 5 grass, cupressus, parietaria and corylus sensitization were encountered in 19.8%, 10.9%, 3.8%, 1.7% and 1.3% respectively. Prevalence of sensitization to pollens didn't vary significatively according to age: 25.4% in females versus 31.1% in males (p = 0.13). Prevalence of pollen sensitization was 26.6% in case of asthma versus 30.9% in non asthmatics patients (p = 0.30), 30.1% in case of rhinitis versus 16.9% in non rhinitics (p = 0.009), 36.4% in case of conjunctivitis versus 22.7% in case without conjunctivitis (p < 0.0002). According to the regions, prevalence was significatively high in areas like Meknes (56%), Beni-Mellal (48.1%), Marrakech (43.5%) where olive is abundant in comparison with Oujda (30.8%), Agadir (30.8%), Tanger (27.3%), Rabat (21.8%), Safi (21.3%), Casablanca (11.6%) and El Jadida (10%). The prevalence was up to 84.3% when symptoms were predominant in spring versus 16.8% in other seasons (p < 0.0001). Thus, although SPT did not include numerous other pollens, olive pollinosis seems to be prevalent in Morocco, followed by allergy to grass pollen. Such studies need to be carried out in line with pollen count. PMID- 11887773 TI - [Inflammatory pseudotumors of the lung with severe course]. AB - Pulmonary inflammatory pseudotumors are usually unique lesions of unknown etiology with good prognosis. We report two severe cases with mediastinal invasion, local recurrence, extrathoracic locations, one of them with a fatal evolution. Certain microscopic features, which were present in our cases (increased cellularity, nuclear pleomorphism, mitotic activity, focal necrosis, bizarre giant cells, vascular invasion), may have prognostic relevance in determining an aggressive behavior of these tumors. Surgical resection is the recommended treatment, and incomplete resection, as in our cases, seems to be a risk factor for developing recurrent inflammatory pseudotumor. Immunosuppressive therapy was ineffective as well as radiotherapy in one of our cases. Only high doses of corticosteroids seemed to slow the evolution of the disease. PMID- 11887774 TI - [Endocarditis, meningitis, pneumopathy and pneumococcal cerebral abscess in an alcoholic smoker]. AB - We report a case of mitral endocarditis caused by Streptococcus pneumoniae in a 43 year old man with history of alcohol abuse and cigarette smoking. The pneumococcal endocarditis was associated with pneumonia, meningitis and brain abscess. Only transesophageal echocardiography could confirm the presence of vegetation. The patient was treated medically with good results. PMID- 11887778 TI - Healthcare online.... Legal research. PMID- 11887776 TI - [Recurrent edema of the face and neck in a chronically hemodialyzed patient]. AB - We report the case of a patient undergoing long-term hemodialysis admitted to hospital for diagnosis of recurrent face and neck edema influenced by dialysis sessions with paroxysmal dyspnea. We considered the possible role of allergy to ethylene oxide and to formaldehyde without diagnostic confirmation. Dialyzer complement activation was suspected but changing the dialyzer did not improve the symptoms. Anti-histaminic and corticosteroid therapy did not modify symptoms. A mild hemithoracic collateral circulation occurred and led to the discovery of a superior vena cava syndrome. Computed tomography and bilateral upper limb contrast venography visualized a thrombus in the superior vena cava extending into the right venous brachiocephalic arm from the central vein catheter. A stent was inserted into the superior vena cava which, together with anticoagulant therapy, led to rapid resolution of the symptoms. Superior vena cava syndrome related to a central catheter and hypersensitivity reactions should always be considered as possible causes of recurrent face and neck edema in patients on long-term hemodialysis. PMID- 11887775 TI - [Allergic bronchopulmonary aspergillosis disclosing mucoviscidosis]. AB - In predisposed patients, allergic bronchopulmonary aspergillosis (ABPA) can arise from aspergillus bronchial colonization. We report the case of a young woman who presented with a right basal pneumonia, ground glass opacities and mediastinal adenopathies on CT scan. Biological, radiological and clinical criteria, as well as an history of childhood asthma, allowed the initial diagnosis of ABPA. However, the unusual coexistence of an additional infection with Pseudomonas Aeruginosa evoked the diagnosis of cystic fibrosis, confirmed by a sweat test and genetic analysis. Under corticosteroid and antifungal therapy and antibiotics, the clinical and radiological evolution was favourable but immuno-allergic sensitisation persisted. The ABPA-cystic fibrosis association is not rare with an estimated prevalence of 2% to 11% according to previous studies. This variability is partly explained by the difficulty of the diagnosis due to confounding clinical, radiological, and biological signs between ABPA and cystic fibrosis. Many predictive development factors of ABPA in the context of cystic fibrosis have been reported, including respiratory function, personal or familial atopy, colonization with Pseudomonas Aeruginosa and age. As in non cystic fibrosis patients, the treatment requires systemic corticotherapy and itraconazole. ABPA is still often under diagnosed and should be evoked in the context of cystic fibrosis. PMID- 11887779 TI - Man's best friend receives man's best healthcare. PMID- 11887782 TI - The nursing shortage: a window of opportunity. PMID- 11887780 TI - Nursing workforce: a perspective for now and the future. AB - The vital links between nursing workforce research and policy are clear. Documented findings on current and projected continued shortages of nurses have lead to new legislation and policies targeted to increase the numbers of qualified nurses and to improve the image of nurses. These initiatives will be effective only if there is concurrent leadership in the practice arena. Research and policy must be linked to practice through initiatives to strengthen leadership, develop new practice models that improve utilization of nurses and patient satisfaction, and strengthen nursing's participation in decision making in healthcare delivery. PMID- 11887781 TI - The nursing shortage--it's back! AB - Nursing shortages have been cyclic events during the 1970s, 1980s, and early 1990s. Recent downsizings and reengineerings have witnessed professional nurses in reduction of workforce actions. Acute-care settings, with goals of increased productivity and cost reduction, have decreased ratios of professional to nonlicensed staff frequently forcing nurses to practice with fewer resources and in diverse healthcare settings. As inpatient settings find a higher acuity level of patients receiving care from a "leaner" high-percentage agency workforce, many experts believe we are beginning to experience a new wave of shortages that will be more critical than those of the past. Hospitals and other healthcare practice sites will experience a shortage of professionals necessary to provide care to patients and will be competing for those scarce resources. This article presents a discussion of the important factors of supply and demand related to the shortage and possible strategies for hospitals to use when approaching the large and complex challenge. PMID- 11887777 TI - [Thoracic pain and balloon image in a young woman]. PMID- 11887783 TI - Mandatory overtime: professional duty, harms, and justice. PMID- 11887784 TI - Display of complete life cycle of human papillomavirus type 16 in cultured placental trophoblasts. AB - Human papillomavirus (HPV) infection is threefold more prevalent in spontaneous abortion specimens compared to elective abortions preferentially targeting the placental trophoblasts in these specimens. Here by using infectious ceplar and Southern blot analysis, we demonstrate that the transfected HPV-16 genome de novo replicates in 3A trophoblasts in culture. Peak DNA replication occurred 9-24 days posttransfection, showing classic DNA forms I, II, and III and an 8-kb monomer band upon DpnI/BamHI digestion. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of mRNA expression revealed that E6 and E2 were significantly expressed by day 9, coinciding with HPV-16 DNA replication. However, significant L1 expression was delayed until day 18. L1 protein expression on day 18, but not day 9, was also confirmed by Western blot analysis. The production of HPV-16 virions was demonstrated by three techniques: the appearance of HPV-16 infectious units coinciding with L1 expression, the neutralization of these infectious units with known neutralizing anti-HPV-16 antibodies, and the appearance of spliced E1 E4 and E6-E7 transcripts (RT-PCR) in normal keratinocyte rafts infected with these trophoblast-produced HPV-16 infectious units. These data suggest that HPV 16 is carrying out its complete life cycle in trophoblasts. Previously, HPVs were known to productively replicate only in differentiating keratinocytes of skin. These findings expand HPV biology, support the hypothesis of a possible link between HPV and some spontaneous abortions, and present a new technology for studying HPV. PMID- 11887785 TI - Proliferation of cerebral cortical cells from the hydrocephalic HTx rat: an in vitro study. PMID- 11887786 TI - The clinic visit does not end when the patient walks out the door. PMID- 11887787 TI - A revitalization and new initiatives at the University of Texas Houston Dental Branch. PMID- 11887788 TI - Tell me exactly when you're going to die...and I'll tell you what kind of life insurance to buy. PMID- 11887789 TI - Retraction. PMID- 11887792 TI - Introduction to CAM. PMID- 11887793 TI - O-glycosylation of the mouse hepatitis coronavirus membrane protein. PMID- 11887790 TI - The minimally conscious state: definition and diagnostic criteria. PMID- 11887794 TI - Transtympanic endoscopy for drug delivery to the inner ear using a new microendoscope. AB - Anatomic variations of the round window (RW) niche found in approximately 33% of human temporal bones may account for some ofthe problems associated with local drug delivery to the inner car. A microendoscope with a total outer diameter of 1.2 mm was developed in particular for easy visualization and of drug delivery to the RW niche. It incorporated a thin fiber optic, a working/laser channel (0.3 mm) and an irrigation/suction channel (0.27 mm). When compared to a common 30 degree lens optic, with the microendoscope a greater area of the round window niche could be overseen. In addition, the endoscope could be advanced directly upon the surface of the RW membrane (RWM). The microendoscope may be used for evaluation of the anatomy of the RW niche prior to the placement of local drug delivery systems, for application of drugs directly onto the surface of the RWM or to verify the correct placement of inner ear drug delivery systems. PMID- 11887797 TI - [Cracow medical chamber in the special collection of the main medical library]. AB - The article on Cracow Medical Chamber in the Special Collection of the Main Medical Library concerns the history of establishing medical chambers and covers the period from 1893 to 1950. It presents in detail the territorial extent of Cracow Medical Chamber, its membership and presidents' functions as well as the composition of its various boards and councils. It also talks about the legal acts regulating the functioning of medical chambers and their sphere of activity. The collection of archived records kept in the Special Collection of Main Medical Library constitutes a small percentage of the preserved records; these being mainly personal files of doctors comprising personal questionnaires, registration cards and photographs. PMID- 11887796 TI - [Convent nursing in Polish hospitals from 1939]. AB - The author, Sister of Charity and Mother Superior of the Children's Home in Tarnow, describes the attitude of the authorities of the Polish People's Republic towards convent nursing in hospitals. The problem is exemplified by the situation of the Cracow Province of the Order of the Sisters of Charity. Apart from this main theme, Sister Miroslawa Halat says a little about the establishment of the Order and the role it played in Polish hospital care. Before presenting the fundamental problems related to the topic of the article, the author presents numerical data showing the Sisters of Charity's involvement in hospital care between World War I and World War II. This facilitates a comparison of the possibilities of providing hospital care by the Sisters of Charity at a later time. The author also describes the persecutions of the Sister of Charity by the German occupiers. Her main reflections are divided into several parts. The period from 1945 to 1949 covers the time when the Sisters of Charity tried to engage actively in the reconstruction of the ruined Polish hospital system. This involved leaving the former Polish eastern borderland and taking up duties in the so-called Regained Territories. The period from 1950 to 1989 describes the conscious, planned and ideologically directed policies of the authorities of the Polish People's Republic to nullify or at least diminish the role of the Sisters of Charity in Polish hospitals. Quoting numerous accounts and documents, the author tries to recreate the atmosphere of those times. She also writes about the Sisters of Charity's preparation for work in hospitals as well as the possibilities of their education after the war. The article presents the Sisters of Charity's activity under very difficult political conditions and is complemented by a great number of figures, graphs and source documents. PMID- 11887795 TI - Design, fabrication, and evaluation of high frequency, single-element transducers incorporating different materials. AB - The performance of high frequency, single-element transducers depends greatly on the mechanical and electrical properties of the piezoelectric materials used. This study compares the design and performance of transducers incorporating different materials. The materials investigated include 1-3 lead zirconate titanate (PZT) fiber composite, lead titanate (PbTiO3) ceramic, poly(vinylidene fluoride) (PVDF) film, and lithium niobate (LiNbO3) single crystal. All transducers were constructed with a 3-mm aperture size and an f-number between 2 and 3. Backing and matching materials were selected based on design goals and fabrication limitations. A simplified coaxial cable tuning method was employed to match the transducer impedance to 50 ohms for the PZT fiber composite and PbTiO3 ceramic transducers. Transducers were tested for two-way loss and -6 dB bandwidth using the pulse/echo response from a flat quartz target. Two-way loss varied from 21 to 46 dB, and bandwidths measured were in the range from 47 to 118%. In vitro ultrasonic backscatter microscope (UBM) images of an excised human eye were obtained for each device and used to compare imaging performance. Both press focusing and application of a lens proved to be useful beam focusing methods for high frequency. Under equal gain schemes, the LiNbO3 and PbTiO3 transducers provided better image contrast than the other materials. PMID- 11887800 TI - Prosthetic conformers: a step towards improved rehabilitation of enucleated children. AB - Enucleation in children is distressing for families, particularly because of concerns of cosmesis. In the last 2 years the authors have used painted conformers instead of clear conformers to make the postoperative healing and rehabilitation period easier on the families. A set of six prosthetic conformers (small, medium and large; blue and brown) was available in the operating room. An appropriately sized and colour-matched conformer was placed in the socket at the end of surgery and kept for an average of 4-6 weeks. This decreased the psychological impact of enucleation, yet achieved the goals of an ideal conformer allowing optimal wound healing without pressure to fit a permanent individualized prosthesis earlier than 6 weeks after surgery. The acceptance of families to prosthetic conformers in this paediatric population has been very positive, improving rehabilitation of the family and the enucleated child. PMID- 11887798 TI - [The Saska Library in Dresden as a research centre for history of medicine]. AB - The article serves a guide to the collection in the Saska Library, related to medicine. PMID- 11887799 TI - [Historical outline of the Medical Department Museum of the Jagiellonian University]. AB - The article presents the history of the Museum of History of the Medical Department of the Jagiellonian University in Cracow, which was founded in 1900. Initiated and created by Walery Jaworski, professor of Internal Diseases, the museum has gathered a rich and varied collection of artifacts. These comprise such exhibits as manuscripts, old texts, medical diplomas as well as photographs of doctors, medical and laboratory equipment and a collection of medals and mementoes of professors from Cracow University. PMID- 11887803 TI - Influence of neutral-pH dialysis solutions on the peritoneal membrane: a long term investigation in rats. AB - OBJECTIVE: Glucose degradation products (GDPs) and low pH are potential causes of bioincompatibility of peritoneal dialysis fluids (PDFs). The aim of the present study was to compare the effect of 6 weeks' exposure of the peritoneum in rats to two different PDFs: a standard PDF with a low pH and high level of GDPs (CAPD 3: Fresenius Medical Care, Bad Homburg, Germany), and a modified PDF with a low level of GDPs and a physiologic pH (CAPD 3 Balance: Fresenius Medical Care). METHODS: After catheter implantation, rats were exposed twice daily for 6 weeks to CAPD 3 fluid or to CAPD 3 Balance. At the beginning and at the end of the study, a 4-hour dwell was performed in every rat to evaluate intraperitoneal inflammation and its effect on total collagen synthesis in the in vitro cultured rat mesothelial cells (ex vivo study). Additionally, after 6 weeks' exposure, the peritoneal cavity was opened, and macroscopic changes were evaluated according to a semiquantitative scale. Peritoneal samples were also taken for morphology study. RESULTS: In rats treated with CAPD 3 fluid, intraperitoneal inflammation was comparable at the beginning and at the end of the experiment. In animals exposed to CAPD 3 Balance, the intensity of the intraperitoneal inflammation decreased during the study (cell count, p = 0.0781; neutrophil:macrophage ratio, p < 0.01; nitrite concentration, p < 0.05; hyaluronan level, p < 0.05). The capacity of effluent dialysate from CAPD 3 rats to activate collagen synthesis in in vitro-cultured mesothelial cells was the same at the beginning and at the end of the study. In the CAPD 3 Balance group, this capacity was statistically significantly lower at the end of the study than at the beginning (p < 0.05). The mean thickness of the visceral peritoneum was comparable in both groups of animals, but, macroscopically, more severe fibrosis was found in the peritoneum of rats exposed to CAPD 3 as compared with animals treated with CAPD 3 Balance (p < 0.05). CONCLUSION: We showed that, in the rat model of peritoneal dialysis, chronic exposure of the peritoneum to PDFs with low GDPs and a physiologic pH diminished the intraperitoneal inflammatory reaction induced by dialysis, and reduced peritoneal fibrosis. PMID- 11887801 TI - A proposed 3D structure for crotamine based on homology building, molecular simulations and circular dichroism. AB - Crotamine, isolated from the venom of the South American rattlesnake Crotalus durissus terrificus is a strongly basic 42-amino acid polypeptide belonging to the small basic myotoxin family. As no tridimensional structure is available for this myotoxin subfamily, despite its important pharmacological interest, we propose in this paper a theoretical 3D model for crotamine. Starting from a homology modelling procedure, followed by intensive molecular dynamics (MD) simulations in water and complementary CD experiments, the designed 3D model is the first example of a tridimensional structure in this family of small basic myotoxins. Crotamine, therefore, belongs to a newly identified structural family presenting a common fold also found in beta-defensin and antopleurine-B. The proposed 3D model will be used for future calculations about crotamine aggregation and interaction with membranes. PMID- 11887804 TI - Biologic significance of reduced levels of glucose degradation products. PMID- 11887806 TI - A food chemist's view of advanced glycation end-products. PMID- 11887805 TI - Glucose degradation products in peritoneal dialysis fluids: how they can be avoided. AB - OBJECTIVES: A patient on peritoneal dialysis (PD) uses 3-7 tons of PD fluid every year. The result is considerable stress on the peritoneal tissue. Aspects of PD fluids that have been considered responsible for bioincompatibility are low pH, high osmolality, high glucose and lactate concentrations, and the presence of glucose degradation products (GDPs). However, the relative importance of each factor in PD fluid has so far not been investigated. Discovering their relative importance was the aim of the present study. METHODS: Two main methods for investigating biocompatibility were used in this study: cytotoxicity measured as in vitro inhibition of cell growth, and in vitro AGE formation measured as albumin-linked fluorescence. RESULTS: The two most important factors for determining in vitro bioincompatibility of PD fluids were the presence of GDPs, which caused both severe cytotoxicity and strong AGE promotion, and low pH, which induced severe cytotoxicity. CONCLUSIONS: The biocompatibility of PD fluids can be monitored through fairly simple in vitro methods such as cell proliferation and AGE formation. Bioincompatibility of PD fluids is caused mainly by the presence of GDPs and low pH. These findings correlate well with known clinical bioincompatibility. PMID- 11887808 TI - Continuous flow peritoneal dialysis. PMID- 11887802 TI - Interleukin-6 levels decrease in effluent from patients dialyzed with bicarbonate/lactate-based peritoneal dialysis solutions. AB - OBJECTIVE: Conventional lactate-buffered peritoneal dialysis (PD) solutions have several bioincompatible characteristics, including acidic pH, lactate buffer, and the presence of glucose degradation products (GDPs). These characteristics, along with inflammation, are believed to contribute to membrane dysfunction in peritoneal dialysis patients. A new PD solution containing a bicarbonate/lactate buffer system with physiologic pH and low GDPs has shown improved biocompatibility in both in vitro and ex vivo studies. In the present study, the concentrations of cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), and vascular endothelial growth factor (VEGF), were measured in timed overnight effluents from PD patients continuously dialyzed with either lactate based control solution (C) or bicarbonate/lactate-based solution (B/L) for 6 months. METHODS: Effluents from 92 continuous ambulatory peritoneal dialysis (CAPD) patients were collected when the patients were entered into the study (baseline, all patients on C for more than 3 months), and at 3 and 6 months following randomization to C (n = 31) or to B/L (n = 61). Effluent samples were filtered, stored frozen, and then assayed for IL-6, TNFalpha, and VEGF by ELISA. RESULTS: A significant decrease in effluent IL-6 was seen at 3 months and at 6 months in the B/L-treated patients. Levels of VEGF were significantly reduced at 3 months. No changes in the levels of IL-6 or VEGF were seen in the C-treated patients, and no changes in TNFalpha were seen in either group over time. CONCLUSIONS: Treatment with B/L is associated with decreased IL-6 synthesis and decreased VEGF secretion. The data suggest that the use of B/L solution is associated with reduced intraperitoneal inflammation and potential for angiogenesis. The use of B/L solution may, over time, help to restore peritoneal homeostasis and therefore preserve the function of the membrane in peritoneal dialysis. PMID- 11887809 TI - Peritoneal dialysis fill volume: can the patient tell the difference? AB - OBJECTIVE: Patients on continuous ambulatory peritoneal dialysis (CAPD) must receive an increased dialysis dose as they lose residual renal function so that total clearances are optimized. The dialysis dose may be increased by increasing the exchange volume. Patients on CAPD are often reluctant to use a greater exchange volume, fearing increased pain and discomfort and an altered body image. To assess patient perception of various fill volumes, we studied 12 stable patients currently treated with 2-L exchanges who had no surgical contraindication to larger fill volumes. METHOD: After an overnight dwell, patients received a 2-L, 2.5-L, or 3-L exchange of Baxter PD4 (Baxter Healthcare SA, Castlebar, Ireland) for 3 hours in a randomized crossover design. Patients and staff were both blinded to the fill volume. At the beginning and end of the exchange, intraperitoneal hydrostatic pressure (IPP) in the supine position was measured, and the patient's perception of the exchange was evaluated using the validated McGill Pain Questionnaire (MPG). RESULTS: Initial IPP increased with increasing fill volume (12.5 +/- 3.7 cmH2O vs 16.1 +/- 4.2 cmH2O vs 18.7 +/- 3.6 cmH2O for 2, 2.5 L, and 3L, respectively). For all fill volumes, IPP had fallen by the end of the 3-hour dwell, at which time it was similar to that after an overnight 2-L exchange. The pain rating index by was generally low for all exchange volumes and did not correlate with IPP. Minor degrees of discomfort were reported by 4, 2, and 1 patients with 3-L, 2.5-L, and 2-L exchanges respectively. CONCLUSION: Our study suggests that, despite an increased IPP, larger exchange volumes are generally well tolerated by patients, with only a minority of patients feeling mild discomfort. PMID- 11887810 TI - Optimization of peritoneal dialysis prescription using computer models of peritoneal transport. AB - Computer models are valuable clinical tools in the effort to improve quality of life for dialysis patients. At present, two software programs have been validated clinically in adult and pediatric populations. They are the Personal Dialysis Capacity (PDC: Gambro Lundia AB, Lund, Sweden) and PD Adequest (Baxter Healthcare Corporation, Deerfield, IL, U.S.A.). Both programs seem to give accurate predictions of small-solute clearance, but the PDC seems to be superior in predicting ultrafiltration volumes. Indeed, the software programs have several important differences that affect their accuracy and, hence, their clinical value. The PDC software introduces the concepts of capillary physiology to the field of peritoneal dialysis. It gives a functional description of the peritoneal membrane of the individual patient. Recently, its "new" area parameter (A0/delta x) was shown to be superior to the peritoneal equilibration test (PET) in predicting transperitoneal exchange. PMID- 11887811 TI - Helicobacter pylori infection: a new cause of anorexia in peritoneal dialysis patients. AB - OBJECTIVE: Helicobacter pylori (HP) infection has frequently been found in dialysis patients. Chronic infections induce overproduction of pro-inflammatory substances. Inflammation has been associated with cachexia and anorexia. We explored the relationship between HP infection, anorexia, and malnutrition in peritoneal dialysis (PD) patients. PATIENTS AND METHODS: The study included 48 clinically stable PD patients divided into four groups: HP+ with anorexia (group I, n = 12); HP+ without anorexia (group II, n = 4); HP- with anorexia (group III, n = 5); and HP- without anorexia (group IV, n = 27). Infection with HP was diagnosed by breath test. Anorexia was evaluated using a personal interview and an eating motivation scale (VAS). The VAS included five questions that are answered before and after eating. The questions concern desire to eat, hunger, feeling of fullness, prospective consumption, and palatability. Biochemical markers of nutrition and inflammation were also determined. RESULTS: At baseline, group I showed lower scores for desire to eat, hunger sensation, prospective consumption, and palatability. They also showed lower lymphocyte counts, prealbumin, transferrin, serum albumin, normalized equivalent of protein-nitrogen appearance (nPNA), and residual renal function (RRF). In addition, the same group showed higher levels of C-reactive protein (CRP) and more sensation of fullness than the remaining groups. In the entire series, we found significant linear correlations between the following markers of nutrition and certain questions on the VAS: albumin with before-lunch desire to eat (r = 0.38, p < 0.05), and prealbumin with before-lunch hunger (r = 0.41, p < 0.05) and after-lunch hunger (r = -0.35, p < 0.05). Negative linear correlations were found between albumin and fullness before lunch (r = -0.45, p < 0.01), and between prealbumin and before-lunch desire to eat (r = -0.39, p < 0.05). Negative linear correlations were also seen between CRP and albumin (r = -0.35, p < 0.05) and between CRP and prealbumin (r = -0.36, p < 0.05). Similarly, CRP showed a negative correlation with before-lunch desire to eat (r = -0.38, p < 0.05) and afterlunch desire to eat (r = -0.45, p < 0.01). After HP eradication, group I showed a significant increase in markers of nutrition and in VAS scores for almost all questions. Simultaneously, they showed a decrease in CRP level. Significant differences were also found in lymphocyte count (1105 +/- 259.4 cells/mm3 vs 1330.8 +/- 316 cells/mm3, p < 0.05), nPNA (0.9 +/- 0.16 g/kg/day vs 1.07 +/- 0.3 g/kg/day, p < 0.05), prealbumin (26.7 +/- 6.5 mg/dL vs 33.9 +/- 56.6 mg/dL, p < 0.01), albumin (3.48 +/- 0.3 g/dL vs 3.67 +/- 0.35 g/dL, p < 0.05), CRP (1.16 +/- 1.14 mg/dL vs 0.88 +/- 1.2 mg/dL, p < 0.054), before-lunch desire to eat (56.6 +/- 6.8 vs 72.2 +/- 4, p < 0.001), after-lunch desire to eat (5.4 +/- 2.6 vs 12.3 +/- 2, p < 0.01), hunger before lunch (55.4 +/- 5.4 vs 73.1 +/- 4.6, p < 0.001), hunger after lunch (5.8 +/- 2.9 vs 11 +/- 4, p < 0.01), fullness before lunch (36.6 +/- 10.3 vs 18.7 +/- 8.8, p < 0.001), consumption after lunch (5 +/- 4.7 vs 17.5 +/- 18, p < 0.05), and palatability (61 +/- 5.3 vs 74.1 +/- 4.1, p < 0.001). CONCLUSION: Infection with HP is associated with anorexia, inflammation, and malnutrition in PD patients. Eradication of HP significantly improves this syndrome. Residual renal function seem to have a protective effect on appetite preservation. The present study supports the hypothesis of the involvement of inflammation in the pathogenesis of malnutrition in PD patients. PMID- 11887807 TI - Adequacy targets can be met in anuric patients by automated peritoneal dialysis: baseline data from EAPOS. AB - OBJECTIVE: Conventional continuous ambulatory peritoneal dialysis (CAPD) in patients without residual renal function and with high solute transport is associated with worse clinical outcomes. Automated peritoneal dialysis (APD) has the potential to improve both solute clearance and ultrafiltration in these circumstances, but its efficacy as a treatment modality is unknown. The European Automated Peritoneal Dialysis Outcomes Study (EAPOS) is a 2-year, prospective, European multicenter study designed to determine APD feasibility and clinical outcomes in anuric patients. The present article describes the baseline data for patients recruited into the study. DESIGN: All PD patients treated in the participating centers were screened for inclusion criteria [urinary output < 100 mL/24 h, or residual renal function (RRF) < 1 mL/min, or both]. After enrollment, changes were made to the dialysis prescription to achieve a weekly creatinine clearance above 60 L per 1.73 m2 and an ultrafiltration rate above 750 mL in 24 hours. SETTING: The study is being conducted in 26 dialysis centers in 13 European countries. BASELINE DATA COLLECTION: The information collected includes patient demographics, dialysis prescription, achieved weekly creatinine clearance, and 24-hour ultrafiltration (UF). RESULTS: The study enrolled 177 anuric patients. Median dialysis duration before enrollment was 22.5 months (range: 0-285 months). Mean solute transport measured as the dialysate-to-plasma ratio of creatinine (D/P(Cr)) was 0.74 +/- 0.12. Patients received APD for a median of 9.0 hours overnight (range: 7-12 hours) using a median of 11.0 L of fluid (range: 6-28.75 L). Median daytime volume was 4.0 L (range: 0.0-9.0 L). Tidal dialysis was used in 26 patients, and icodextrin in 86 patients. At baseline, before treatment optimization, the weekly mean total creatinine clearance was 65.2 +/- 14.4 L/1.73 m2, with 105 patients (60%) achieving the target of more than 60 L/1.73 m2. At baseline, 81% of patients with high transport, 69% with high-average transport, and 40% with low-average transport met the target. At baseline, 70% of patients with a body surface area (BSA) below 1.7 m2, 60% with a BSA of 1.7-2.0 m2, and 56% with a BSA above 2.0 m2 achieved 60 L/1.73 m2 weekly. Median UF was 1090 mL/24 h, and 75% of patients achieved the UF target of more than 750 mL/24 h. CONCLUSION: This baseline analysis of anuric patients recruited into the EAPOS study demonstrates that a high proportion of anuric patients on APD can achieve dialysis and ultrafiltration targets using a variety of regimes. This 2-year follow-up study aims to optimize APD prescription to reach predefined clearance and ultrafiltration targets, and to observe the resulting clinical outcomes. PMID- 11887813 TI - The measurement of total body potassium in patients on peritoneal dialysis. AB - OBJECTIVES: To assess the validity of measuring total body potassium (TBK) to estimate fat-free mass (FFM) and body cell mass (BCM) in patients on peritoneal dialysis (PD). METHODS: We studied 29 patients on PD (14 men, 15 women) and 30 controls (15 men, 15 women). We calculated TBK by using a whole-body counter to measure 1.46 MeV gamma-ray emissions from naturally occurring 40K. We measured total body water (TBW) by deuterium oxide dilution, and extracellular water (ECW) from bromide dilution. These measurements allowed us to estimate intracellular water (ICW), fat-free mass dilution (FFM(Dilution)), and body cell mass dilution (BCM(Dilution)). RESULTS: The FFM(TBK) in male PD patients (55.7 +/- 7.0 kg) did not differ from that in male controls (57.0 +/- 10.9 kg). The FFM(TBK) in female PD patients (38.4 +/- 6.8 kg) was less than that in female controls (44.7 +/- 4.5, p < 0.01). The FFM(Dilution) did not differ from the FFM(TBK). Correlation of FFM(TBK) and FFM(Dilution) was r = 0.90, p < 0.0001 for all subjects; r = 0.90, p < 0.0001 for PD patients; and r = 0.90, p < 0.0001 for controls. Bland Altman comparison of FFM(Dilution) with FFM(TBK) in individuals showed bias 0.6 kg, range -8.5 kg to 9.7 kg for the whole group; bias 1.4 kg, range -7.9 kg to 10.7 kg for PD patients; and bias -0.2 kg, range -9.0 kg to 8.6 kg for controls. The BCM(TBK) in male PD patients (30.1 +/- 4.5 kg) did not differ from that in male controls (31.9 +/- 6.2 kg). The BCM(TBK) in female PD patients (19.0 +/- 4.4 kg) was less than that in female controls (23.1 +/- 2.9 kg, p < 0.01). The BCM(Dilution) results did not differ from those for the BCM(TBK). Correlation of BCM(TBK) and BCM(Dilution) was r = 0.90, p < 0.0001 for all subjects; r = 0.87, p < 0.0001 for PD patients; and r = 0.93, p < 0.0001 for controls. Bland-Altman comparison of BCM(Dilution) with BCM(TBK) in individuals showed bias 0.1 kg, range -5.9 kg to 6.1 kg for the whole group; bias 0.0 kg, range -6.9 kg to 6.9 kg for PD patients; and bias 0.1 kg, range -5.0 kg to 5.2 kg for controls. The [K+]ICW did not differ between PD patients and controls (148.0 +/- 25.1 mmol/L vs 148.1 +/- 14.3 mmol/L, p = nonsignificant). CONCLUSIONS: Total body potassium is a valid, noninvasive technique for measuring FFM and BCM in PD patients. In our PD patient group, depletion of FFM and BCM as compared with controls was identified in the women but not in the men. PMID- 11887812 TI - Malnutrition, inflammation, and atherosclerosis in peritoneal dialysis patients. PMID- 11887814 TI - Is there a relationship between diet and nutrition status in continuous ambulatory peritoneal dialysis patients? PMID- 11887815 TI - Should the DOQI adequacy guidelines be used to standardize peritoneal dialysis in children? AB - The complete set of PD adequacy guidelines generated by NKF-DOQI is a valuable asset to the care of pediatric and adult patients receiving PD. Use of the recommendations generated by an evidence-based review of the literature (in addition to the expert opinion of the workgroup members who served as authors) has undoubtedly resulted in greater attention to detail in patient care and a more uniform approach to therapy. Whether the target clearances advocated for adults should serve as the standard for children is an issue where evidence is scant. Pertinent data needs to be generated by the pediatric nephrology community. Large pediatric study groups such as the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS), the Pediatric Peritoneal Dialysis Study Consortium (PPDSC), and the Mid-European Pediatric Peritoneal Dialysis Study Group (MEPPS) need to collect information on dialysis clearance and a host of outcome variables such as hospitalization rate, technique survival, nutrition status, statural growth, neurodevelopment, quality of life, and patient mortality. The same groups should also encourage the development of prospective studies designed to further improve the dialysis care of children. Finally, it should be evident to all clinicians and emphasized in the DOQI guidelines that PD adequacy in children is defined by more than just solute and fluid control. Only when anemia, nutrition, infection, growth, and quality of life are optimally managed in combination with effective dialysis can a clinician feel confident that the child on PD has been provided with needed and deserved care. PMID- 11887816 TI - Is growth a valid outcome measure of dialysis clearance in children undergoing peritoneal dialysis? AB - OBJECTIVE: Our study evaluated growth as a clinical outcome measure of peritoneal dialysis (PD) adequacy in children with end-stage renal disease (ESRD). DESIGN: This retrospective single-center study was carried out in our tertiary-care medical center. PATIENTS: The study enrolled 24 patients who initiated dialysis after January 1, 1995, and who had been on dialysis for a minimum of 1 year. RESULTS: The weekly mean total [PD + residual renal function (RRF)] creatinine clearance (C(Cr)) and Kt/V(urea) were 70.3 +/- 18 L per 1.73 m2 and 3.45 +/- 0.73, respectively. Of the 24 patients, 12 (50%) were anuric. The mean height standard deviation score (SDS) changed to -1.78 at the end of 1 year from -1.58 at baseline. Catch-up growth (positive delta height SDS) was observed in 9 patients (37%), 7 of whom (78%) had residual renal function (RRF). In contrast, only 5 of 15 patients (33%) with a negative deltaSDS for height had RRF (p < 0.025). The mean height SDS in patients with RRF improved to -1.64 from -1.78; in patients without RRF, it worsened to -1.90 from -1.37 (p = 0.01). While the weekly total Kt/V(urea) in patients with RRF (3.53) was similar to that in patients without RRF (3.37, p = 0.6), only the native Kt/V(urea) had a significant (but weak) positive correlation with delta height SDS (r2 = 0.17, p = 0.04). In contrast, the total weekly C(Cr) was significantly higher (p = 0.001) in patients with RRF (81.1 L/1.73 m2) as compared with those without RRF (59.5 L/1.73 m2). However, only the native C(Cr)--and not the dialysis C(Cr)--had a significant (but weak) positive correlation with delta height SDS (r2 = 0.17, p = 0.04). CONCLUSIONS: These preliminary data provide evidence for a correlation between solute clearance and growth, with RRF exerting a significant influence on that outcome. The Kt/V(urea) data also appear to contradict the presumed equivalence of PD and native clearance in children with ESRD. PMID- 11887817 TI - Should the DOQI adequacy guidelines be used to standardize the peritoneal dialysis dose in children? PMID- 11887818 TI - Molecular mechanisms of peritoneal permeability--research in growth factors. PMID- 11887819 TI - The DOQI pediatric nutritional guidelines--critical remarks. PMID- 11887820 TI - Peritonitis in automated peritoneal dialysis: antibiotic therapy and pharmacokinetics. PMID- 11887822 TI - Implantation of presternal catheter using Moncrief technique: aiming for fewer catheter-related complications. AB - OBJECTIVES: To reduce catheter-related complications, we developed a new technique of catheter implantation, combining a presternal catheter with the Moncrief technique. METHODS: The presternal catheter, consisting of 2 catheters joined by a titanium extender, was surgically implanted. Its end was left embedded in the presternal wall. A few weeks after implantation, the embedded subcutaneous catheter was exteriorized, exiting in the 4th intercostal space, and peritoneal dialysis (PD) was commenced. RESULTS: Using the new technique, 9 catheters were implanted (3 in women and 6 in men). Exteriorization was performed 30.6 +/- 14.3 days after implantation of the catheter. Total observation period was 70 patient-months. Average hospitalization was 4.4 +/- 1.3 days for catheter implantation, and 2.6 +/- 2.6 days for exteriorization. Peritoneal dialysis commenced on the day of exteriorization with an exchange volume of 1.8 +/- 0.3 L, using 4 exchanges daily. During the observation period, none of the patients experienced a catheter infection or dialysate leak. One non infectious complication was observed (a catheter wrapped in omentum). CONCLUSIONS: Our approach of combining a presternal catheter and the Moncrief technique had some advantages not only in regard to catheter infection and dialysate leakage, but also in regard to quality of life and hospitalization for the patient. PMID- 11887821 TI - Tuberculous peritonitis in a cohort of continuous ambulatory peritoneal dialysis patients. AB - Among 155 patients who were initiated on continuous ambulatory peritoneal dialysis (CAPD), 4 patients (2 men, 2 women) developed tuberculous peritonitis. They had been on PD for between 2 months and 84 months when they developed the peritonitis. The Mantoux test was negative in all of them. The diagnosis was made by a variety of means in the various cases: demonstration of Mycobacterium tuberculosis in the peritoneal cavity; presence of caseating granuloma in a peritoneal biopsy; Mycobacterium tuberculosis in a cold abscess adjacent to the peritoneal cavity; and demonstration of IS6110 and MPB64 genes of Mycobacterium tuberculosis by polymerase chain reaction (PCR) technique. Two of the patients developed ultrafiltration failure. Among 3 patients who were switched to hemodialysis, 2 died and 1 continues on maintenance dialysis. The last patient, whose catheter was removed, was reimplanted with a new catheter and continues on PD without ultrafiltration failure. Any patient with peritonitis unresponsive to conventional therapy should be investigated for tuberculous peritonitis. Institution of chemotherapy without delay will preserve peritoneal membrane integrity. PMID- 11887824 TI - Biofilms in peritoneal dialysis. AB - Microbial adhesions and biofilm bacterial growth have been implicated in serious infections associated with the use of bioprosthetic medical devices and indwelling catheters in humans. Biofilm bacterial growth also commonly occurs on peritoneal dialysis (PD) catheters from skin bacteria. Mature biofilms develop high antibiotic resistance and cause recurrent peritonitis and catheter loss in a subgroup of PD patients. That subgroup of patients can be identified by comparing the antibiotic sensitivities [minimum inhibitory concentrations (MICs)] of a biofilm culture and a routine microbiologic (planktonic) culture of the same PD effluent. PMID- 11887823 TI - Lower malfunction rate with self-locating catheters. AB - OBJECTIVE: We analyzed malfunction rates (obstruction, omental wrapping, displacement) and catheter survival for self-locating catheters as compared with other Tenckhoff catheter designs. PATIENTS AND METHODS: We conducted our survey at two centers, prospectively studying all self-locating catheters implanted from May 1997 to October 2000 and used for peritoneal dialysis (PD). Tenckhoff catheters of other designs used previously in our units were used as the control group. We analyzed removal causes and catheter survival. RESULTS: We studied 173 catheters (105 self-locating catheters, 53 straight catheters, and 15 coiled catheters) implanted in 139 patients (43% of them women) with a mean age of 53 +/ 14 years. The analysis of catheter removal showed that 3 of 105 self-locating catheters, 3 of 15 coiled catheters, and 17 of 53 straight catheters were removed owing to malfunction (chi2: p = 0.0000). Kaplan-Meier curves showed that the bulk of removals for malfunction occurred within the first 3 months after PD start. The group of self-locating catheters showed better survival (log-rank: p = 0.0009). Other causes for catheter removal included peritonitis (n = 22), exit site infection alone (n = 4), and end of PD treatment (n = 66). No significant differences were seen in the annual peritonitis rate (straight-tip: 0.955 +/- 2.315 episodes annually; coiled-tip: 0.651 +/- 0.864 episodes annually; self locating: 0.720 +/- 1.417 episodes annually; t-test: p > 0.400). No gut or bladder perforations were observed. CONCLUSION: In our survey, self-locating catheters were associated with better survival and fewer removals for malfunction than were Tenckhoff catheters of other designs. PMID- 11887833 TI - Building the evidence in peritoneal dialysis: use of randomized controlled trials, and observational and registry data. AB - Renal replacement therapy (RRT) has achieved widespread acceptance without being subjected to the rigors of randomized controlled clinical trials (RCCTs). The RCCT remains the "gold standard" of evidenced-based medicine, but ethical, logistic, and financial limitations mean that not all questions are amenable to a RCCT. Renal registries collect, aggregate, analyze, and interpret data on the occurrence and outcome of renal failure in a defined population. Observational data can be used only to show associations, not causality. Nevertheless, most clinical practice guidelines in nephrology are derived from observational data. The nephrology community needs to join forces to decide the questions that deserve the time, energy, and resources of an RCCT. Prospective observational data can be enhanced by collaboration, standardized definitions, development of a risk-adjustment tool, and consensus among the key players, including professional associations, government, industry, and hospitals. The challenge is to provide evidence-based practice guidelines for the delivery of care to the end-stage renal patient. PMID- 11887828 TI - Aggressive therapy of congestive heart failure and associated chronic renal failure with medications and correction of anemia stops or slows the progression of both diseases. AB - The prevalence of congestive heart failure (CHF) is increasing rapidly in the community. We and others have shown that the prevalence and severity of both anemia and chronic renal failure (CRF) increase steadily with increasing severity of CHF. We have also shown that CHF patients may be resistant to standard drug therapy for CHF as long as the associated anemia is not corrected, and that correction of the anemia with subcutaneous erythropoietin and intravenous iron sucrose (Venofer: Vifor International, St. Gallen, Switzerland) may improve both the CHF and CRF and markedly reduce hospitalizations without causing side effects. We report here our experience with correcting anemia in this manner in 126 cases of anemic-resistant CHF patients. As in our previous studies, correction of the anemia improved both CHF and CRF, and reduced hospitalizations. Our studies suggest that correction of even mild anemia in CHF may be an important addition to the treatment of patients with the combination of CHF and CRF. PMID- 11887834 TI - Integrated care. AB - The integrated care model for delivering RRT offers apparent advantages in patient choice, optimal survival, and economical use of resources. Dialysis and transplantation have always been used that way; but, by integrating all three treatments--HD, PD, and transplantation--the options for the multi-professional team and the patients are increased. The practical implications of the approach include the need for all treatments to be available, for patients and professionals to be educated in the approach, and for a change in therapy to be anticipated and accepted, not seen as a mark of treatment failure. Prospective studies are needed to quantify the benefits in a more systematic way. PMID- 11887830 TI - Current issues in the management of secondary hyperparathyroidism and bone disease. AB - Recent research has demonstrated active bone-like remodelling of vascular tissue in dialysis and non dialysis patients. Cross-sectional studies indicate that the presence of vascular calcification is inversely related to bone mass. Theoretically, the relationship implies that maintaining normal bone turnover and mass may help to decrease vascular calcification. In addition, it is now apparent that phosphorus and the Ca x P product need to be kept as close to normal as possible. Thus, the present goal should be a serum phosphorus of 3.5-5.5 mg/dL, a Ca x P product of <55 mg/dL, and a PTH of around 150-200 pg/mL with the intact assay. Ten years ago, those goals would have been impossible. However, new pharmacologic agents will make their achievement much more realistic. PMID- 11887827 TI - Oral versus intravenous iron supplementation in peritoneal dialysis patients. AB - The vast majority of erythropoietin (EPO)-treated peritoneal dialysis (PD) patients require iron supplementation. Most authors and clinical practice guidelines recommend primary oral iron supplementation in PD patients because it is more practical and less expensive. However, numerous studies have clearly demonstrated that oral iron therapy is unable to maintain EPO-treated PD patients in positive iron balance. Once patients become iron-deficient, intravenous iron administration has been found to more effectively augment iron stores and hematologic response than does oral therapy. We recently performed a prospective, cross-over trial in 28 iron-replete PD patients and showed that twice-monthly outpatient iron polymaltose infusions (200 mg) were a practical and safe alternative to oral iron. That treatment produced significant increases in hemoglobin concentration and body iron stores. The additional expense of intravenous iron therapy was completely offset by reductions in EPO dosage. Careful monitoring of iron stores is important in patients receiving intravenous iron supplementation in view of epidemiologic links with infection and cardiovascular disease. Nevertheless, a growing body of evidence suggests that, as has been found for hemodialysis patients, intravenous iron therapy is superior to oral iron supplementation in EPO-treated PD patients. PMID- 11887826 TI - Hyporesponsiveness to anemia therapy--what are we doing wrong? AB - Most patients receiving anemia therapy respond well, with a significant rise in hemoglobin concentration. However, approximately 5%-10% of patients fail to show a satisfactory response despite high doses of erythropoietin. The definition of hyporesponsiveness to anemia therapy is somewhat arbitrary, but it is generally regarded as a failure to achieve a hemoglobin concentration of 10-11 g/dL despite a dose of erythropoietin in excess of 200 U/kg weekly. The condition has many causes, the most important ones being iron deficiency, infection or inflammation, and underdialysis. Investigating a patient's poor response to erythropoietin should begin with a check for compliance, followed by screening for iron deficiency. If doubt exists about the presence of iron deficiency, then a trial of intravenous iron may be given. A reticulocyte count may be helpful. A significantly elevated count suggests the presence of blood loss or hemolysis. The level of C-reactive protein (CRP) may be useful as an indicator of underlying inflammation, and underdialysis may be corrected by increasing the dialysis prescription. If other, minor causes of hyporesponsiveness to erythropoietin have been excluded, then a bone marrow biopsy should be performed. Some patients may require higher doses of erythropoietin, and it is not unreasonable to increase the dose to 10,000 U thrice weekly. Some causes of hyporesponsiveness to erythropoietin, such as iron deficiency and underdialysis, are easily corrected; but others, such as primary bone marrow disorders and hemoglobinopathies, are not possible to overcome. PMID- 11887825 TI - Peritoneal access in children. AB - Catheter-related infections continue to be the most common complication of CPD, and the most frequent cause of catheter removal. Available evidence supports the superiority of double-cuff catheters and a downward-facing tunnel for preventing peritonitis in children. The Swan-neck double-cuff catheter seems best suited to achieving those objectives, while still reducing the problems of external cuff extrusion and catheter migration. Clearly further pediatric experience with that catheter is desirable. Analysis of the literature confirms that excellent catheter survival and a reduced rate of infectious complications can be achieved with a variety of catheter designs and implantation techniques. The most crucial aspect of catheter success and survival appears to be the commitment and expertise of the team involved in catheter insertion and postoperative catheter management. PMID- 11887835 TI - Factors governing cardiovascular risk in the patient with a failing renal transplant. AB - Cardiomyopathy and IHD are important morbid complications among renal transplant recipients. Age, diabetes, and sex remain important markers of risk. Smoking, hyperlipidemia, and hypertension appear to be the major reversible risk factors for IHD. Anemia and hypertension predict CHF. Definitive evidence on optimal intervention is lacking. Similarities in the renal transplant recipients to CRI patients with respect to cardiomyopathy and to the general population with respect to IHD suggest that extrapolation from those groups is reasonable in the interim. PMID- 11887831 TI - Fragility fractures in dialysis and transplant patients. Is it osteoporosis, and how should it be treated? AB - The term "osteoporosis" must be applied with caution to the uremic population, which has a complex range of metabolic bone disease. Trials of therapeutic interventions to prevent fractures in non uremic populations with osteoporosis cannot be generalized to uremic patients. It is unclear what, if any, role systematic bone densitometry measurement can play in the management of uremic patients who suffer "fragility" fractures--either for diagnostic purposes or to determine the effectiveness of therapy. Estrogen therapy--and perhaps SERMs (raloxifene)--appear to be a reasonable addition to conventional management of secondary HPT with calcium salts and vitamin D analogs. Using bisphosphonates to manage patients who have pre-existing fractures should be considered experimental at best. In certain circumstances, such treatment may be harmful. While the evidence is better that early therapy with intravenous pamidronate in the peri transplant interval may mitigate the steroid-induced bone loss seen in those patients during the first 12 postoperative months, even that indication needs to be subjected to systematic clinical studies to develop appropriate clinical practice guidelines. PMID- 11887832 TI - Bisphosphonates in dialysis and transplantation patients: efficacy and safety issues. AB - Bisphosphonates are an old class of compounds. They were used in the 1930s as antiscaling and anticorrosion agents in washing powders and water to prevent the deposition of calcium crystals. Those basic functions were later utilized in an attempt to prevent ectopic calcifications in humans. The early studies demonstrated that bisphosphonates had a strong affinity for bone. That property was first exploited when the compounds were used for "bone scans." Currently, the drugs are used for treatment of hypercalcemic conditions, abnormal bone remodelling, Paget disease, malignancy, and osteoporosis. Bisphosphonates have several important toxicities: acute renal failure, worsening renal function, reduced bone mineralization, and osteomalacia. For those reasons and others, this class of drugs has not yet been approved for use in children or in patients with severe renal insufficiency. The present review covers several aspects of bisphosphonates: molecular structure, routes of administration, pharmacology, mechanisms of action, toxicities, and exceptional uses in children with renal disease. PMID- 11887829 TI - The role of extracellular matrix in transperitoneal transport of water and solutes. AB - OBJECTIVE: To define the extracellular matrix (ECM), to discuss the physical properties of its components and their impact on transport, and to review data in humans and in animals on the importance of hyaluronan to peritoneal dialysis. METHODS: Literature survey. RESULTS: The ECM fills the interstitium between parenchymal cells and blood vessels in the subperitoneal interstitium. It is responsible for the interstitial resistance to solute and water transfer through the peritoneal barrier. Major components are collagen and hyaluronan, which are synthesized locally in the peritoneal tissue. Synthesis and deposition of these components increase with inflammation, and concentrations of the components influence the mechanical properties of the tissue and the interstitial Starling forces as well as transport. Removal of hyaluronan appears to increase the rates of water and large-solute transport. Addition of hyaluronan to dialysate appears to enhance fluid recovery and to reduce protein loss. CONCLUSION: Many of the physicochemical properties of ECM components are well described, but a large knowledge gap remains concerning the in vivo consequences of specific alterations in the interstitial components. More research is needed. PMID- 11887838 TI - Conception and pregnancy in peritoneal dialysis patients. PMID- 11887837 TI - Undertreatment of cardiac risk factors in adolescents with renal failure. AB - Cardiovascular disease (CVD) is the most common cause of death in adults with end stage renal disease and after renal transplantation, and the relative excess of mortality is greatest in the young. The most likely explanation is the dramatic accumulation of both classical and uremic risk factors leading to atherosclerosis, uremic vasculopathy, and uremic cardiomyopathy. Prospective studies have established the significance of classical and uremic risk factors for the occurrence of CVD in the normal population and in the population with chronic renal disease alike. However, whether and to what degree modification of risk factors by therapeutic intervention can lower morbidity and mortality rates is as yet unknown. PMID- 11887836 TI - Peritoneal dialysis in the patient with a failing renal allograft. AB - BACKGROUND: Patients returning to dialysis treatment after a period with a functioning allograft represent a special case in the integrated care model of renal replacement therapy. They are known to nephrologists--and thus are ideal candidates for a timely commencement of dialysis--but they have had time to accrue additional cardiovascular risk. Furthermore, they have been exposed to prolonged immunosuppression. PURPOSE: The present study aimed to establish the clinical outcomes of patients returning to peritoneal dialysis (PD) with failing allografts [survival, technique survival, longitudinal residual renal function (Kt/V(R)), peritoneal membrane function (solute transport), and plasma albumin] and to compare those outcomes with outcomes in new, contemporary patients. SETTING: The study was conducted in a single center where prospective collection of data now known to be important in determining outcome on peritoneal dialysis (age, comorbidity, albumin, Kt/V(R), and solute transport) has been performed since 1989. METHODS: All patients commencing PD between 1989 and 2001 after failure of an allograft that had functioned for more than 6 months were identified. Outcomes in that group were compared to outcomes in all new PD patients and in all dialysis patients with failed grafts who were returning to hemodialysis (HD) in the same period. RESULTS: The study identified 45 patients with failed allografts: 28 were commencing PD treatment, and 17 were commencing HD treatment.Those patients were significantly younger than the 469 new patients commencing PD [FailedTx-->PD 41.2 years, FailedTx-->HD 38.9 years, NewPD 54.7 years; analysis of variance (ANOVA): p < 0.001]. We saw no significant difference in the survival of failed transplant patients commencing PD as compared with those commencing HD (log rank: p = 0.11). Kaplan-Meier plots of patient survival were better for failed transplant patients as compared with all new PD patients. When corrected using Cox regression, the survival advantage was seen to be due to age and comorbidity at start of PD. Pure technique failure (excluding death) was not different between the groups. Compared with all new PD patients, patients with failed allografts had similar longitudinal plasma albumin and a tendency toward an earlier increase in solute transport, but a more rapid loss of Kt/V(R), (p < 0.05 at 6-48 months). CONCLUSIONS: Peritoneal dialysis would appear to be a good option for patients with failing allografts. Comorbidity is the predominant determinant of survival. That finding underlines the need for attention to factors that might prevent accrual of cardiovascular risk during the post transplantation period. The earlier loss of Kt/V(R) in those patients might be prevented by developing strategies of continued immunosuppression after commencement of dialysis, although infection risk is an important issue. PMID- 11887841 TI - Quality of life of elderly patients undergoing continuous ambulatory peritoneal dialysis. AB - OBJECTIVE: The aim of the present study was to evaluate the impact of continuous ambulatory peritoneal dialysis (CAPD) on the lifestyle of elderly patients. PATIENTS AND METHODS: Aspects of health-related quality of life (OL) were studied in 48 patients (16 men, 32 women) in end-stage chronic renal failure (ESRF) undergoing CAPD at the Clinic of Nephrology, Clinical Centre of Serbia. The first group comprised 20 adult patients (8 men, 12 women; age range: 35-59 years). The second group consisted of 28 older adult patients (8 men, 20 women; age range: 65 75 years). Mean length of CAPD treatment was 5.2 years in the first group and 3.67 years in the second group. Fifteen QL variables were investigated: marital status, family relationships, working ability, sleep, tiredness, appetite, wound healing, hobby, sports, friendships, sexual activity, mood, travel, self management, and happiness. RESULTS: The results showed that, in the examined groups, marital status and relationships with family members weren't influenced at all by dialysis. In both groups, CAPD had a negative influence on ability to bear cold and to travel, but other life functions were not significantly affected. Elderly patients had a significantly worse appetite (p = 0.03, Fisher test) and mood (p = 0.045, Fisher test) than did younger adults. In other examined variables, no statistically significant differences were found between the groups. CONCLUSIONS: Lack of large, statistically significant differences between the groups suggests that CAPD has an equal influence on quality of life in younger and older adult patients. PMID- 11887843 TI - The cost barrier to renal replacement therapy and peritoneal dialysis in the developing world. AB - The socioeconomic statuses of developing nations are diverse. Government policies for reimbursement or coverage of treatment for end-stage renal failure vary greatly from one country to another. Clearly, treatment rates correlate with the gross domestic product and the decision-making process for choosing a specific renal replacement modality is highly influenced by non medical factors. Particularly, those non medical factors include cost issues and the availability of medical and technical resources. Developing nations are experiencing an "epidemiologic transition" as birth rates drop and their populations age. That change in population demographics will undoubtedly increase even further the number of patients at risk of developing end-stage renal disease. Those nations will need to summon the commitment to devote a higher percentage of their resources to treating end-stage renal disease. Finally, when a modality is clearly better for a particular patient, medical indications should play a larger role in the final decision. That approach may prove to be highly useful not only in terms of cost effectiveness and long-term results for the health system, but also in terms of a better quality of life for the patient. PMID- 11887844 TI - A retrospective survey of attitudes toward acceptance of peritoneal dialysis in Chinese end-stage renal failure patients in Hong Kong--from a cultural point of view. AB - OBJECTIVE: We undertook to study the attitudes toward dialysis of patients approaching end-stage renal failure and to analyze those attitudes from a cultural perspective. SETTING: The study was performed in the pre-dialysis clinic of a tertiary referral renal center. PATIENTS: All patients of Chinese ethnic origin seen in the pre-dialysis clinic from 1995 to 2000 for assessment of dialysis therapy were included. METHOD: We performed a retrospective analysis of patient records with regard to attitudes of the patients toward dialysis, reasons for those attitudes, and factors that could lead to a subsequent change in attitude. RESULTS: We assessed 462 patients over the 6-year period. Their mean age was 65.5 +/- 13.3 years, and 43.9% of the patients had diabetes. Peritoneal dialysis (PD) was offered to 74% of the patients, and hemodialysis (HD) to 3.9%. Among the patients offered PD, only 44% accepted dialysis. After counselling, 54% of the patients who originally declined PD ultimately accepted it. The major reasons for refusing PD were the ideas of "having lived long enough" and "lack of family support." Most other reasons could be overcome by counselling. Only a minority of patients demanded hemodialysis. CONCLUSIONS: Declining an offer of dialysis was common. Counselling helped patients to accept PD. Certain cultural elements that hindered acceptance of dialysis were involved in the ideas of "having lived long enough" and "lack of family support." Those cultural elements should be tackled more specifically during counselling. PMID- 11887839 TI - Gastroesophageal reflux and hyperacidity in chronic renal failure. AB - The prevalence of H. pylori infection and PUD seem not to be different in CRF patients as compared with the general population. However, PUD in CRF patients seems to have some unique features-namely, lack of pain and higher associations with bleeding, with post-bulbar location, and with multiple ulcers. No increase in GERD has been proven in adults, but several studies demonstrate increased GERD in pediatric CRF patients. The causes of the increase in GERD may include delayed gastric emptying owing to altered myoelectric activity, or perhaps to an increased production of gastric acid, but evidence for the latter is small. Importantly, treating the problem may lead to better nutrition and higher albumin levels, thus improving patient prognosis. PMID- 11887840 TI - Aquaporins are unlikely to be affected in marked ultrafiltration failure: results from a computer simulation. PMID- 11887845 TI - Vegetarianism in India. PMID- 11887846 TI - The challenges of treating children with renal failure in a developing country. PMID- 11887848 TI - How can we standardize peritoneal thickness measurements in experimental studies in rats? AB - OBJECTIVE: The various methods of measuring peritoneal thickness in experimental studies in rats have yielded conflicting results. Also, no standard method exists to assess histologic findings in peritoneal morphology. We therefore undertook the present study to create a reproducible and standard method for assessing rat peritoneal histology in experimental studies. METHODS: Parietal peritoneal samples from 61 Wistar albino rats were used in the study. Excepting the skin, the whole abdominal wall from each rat was cut two-dimensionally (longitudinally and horizontally), fixed in formalin, and processed routinely for light microscopy. Slides were divided into two groups according to the direction of the inner abdominal muscle fibers in the sections. Longitudinal and horizontal sections of abdominal muscle were evaluated. For every section, one histopathology image was captured from a light microscope to an IBM-compatible computer. Peritoneal thickness (mean of the maximum and the minimum) and submesothelial area (SMA) were drawn on the image. A computer program then automatically performed measurements. Two different measurement methods were compared, based on the same sections. RESULTS: The mean peritoneal thickness was 91 +/- 8 microm in the longitudinal sections and 75 +/- 7 microm in the horizontal sections (p < 0.05). Measurements of the SMA were found to be 47,762 +/- 4,374 microm2 for the longitudinal sections and 40,389 +/- 3,631 microm2 for the horizontal sections (p < 0.05). In both types of sections, a positive correlation (96% for longitudinal and 90% for horizontal) was found between the SMA and the peritoneal thickness (p < 0.01). The SMA measurements correlated significantly with functional properties [ratio of the dialysate concentration of glucose initially and after a 1-hour dwell (D1/D0 glucose), ultrafiltration, and protein loss; p < 0.01]. CONCLUSION: Peritoneal thickness can be measured as a mean of the minimum and maximum values. That measurement strongly correlates with submesothelial area. Both types of sections can be used, but the horizontal and longitudinal sections show systematic differences. All samples in a study should be taken using the same section pattern, either longitudinal or horizontal. PMID- 11887849 TI - Morphologic aspects of chronic peritoneal dialysis in a rat model. AB - OBJECTIVE: We previously put forward an experimental model for evaluating peritoneal function in chronically dialyzed rats. In the present paper, we show the morphologic alterations detected by electron microscopy in the peritonea of chronically dialyzed rats. MATERIALS AND METHODS: The study was done in male Wistar rats. Animals were divided into two groups: control (non dialyzed) rats and dialyzed rats [intraperitoneally exposed to Dianeal 3.86% (Baxter Healthcare Corporation, Deerfield, IL, U.S.A.) for 1 month]. At the end of the study, the rats were humanely killed by bleeding, and samples of the visceral peritoneum covering the liver were taken from 3 rats in each group. The samples were examined by transmission electron microscopy (TEM). RESULTS: In control rats, the peritoneum was composed of flattened mesothelial cells covering the thin layer of the submesothelial connective tissue (ST), containing few fibroblasts. In dialyzed animals, a dramatic expansion of the ST was found. We saw compartmentalization of the ST, with changed morphology of the fibroblasts, altered organization of collagen fibers, and changes in the cells infiltrating the ST. Apart from the fibroblasts, mast cells were relatively numerous. CONCLUSIONS: Our work underlines the capabilities of morphology studies in an animal model for assessing peritoneal dialysis fluid biocompatibility. PMID- 11887847 TI - In vivo peritoneal chamber: linking structure and transport function. AB - OBJECTIVE: The present study aimed to develop an animal model of chronic peritoneal exposure that directly links transport with the tissue involved. METHODS: Daily, rats were intraperitoneally infused through subcutaneous ports with 20 mL of these solutions: isotonic Krebs (K), K + 2.5% mannitol (M), K + 2.5% N-acetylglucosamine (NAG). Controls included catheter-only (CC) and age control rats (AC). After 2 months, each rat was anesthetized and a plastic chamber was affixed to the abdominal wall serosa to isolate a portion of the peritoneum for transport studies. In the first 90 minutes, a hypertonic solution (approximately 500 mosm/kg) containing 14C-mannitol was placed in chamber. The volume and 14C concentration were measured to determine the rate of osmotic flux (flow/Area(chamber)) into the chamber and the flux of mannitol from the chamber to the tissue. At 90 minutes, fluorescein isothiocyanate conjugate (FITC)-albumin was given intravenously. The rate of appearance of that substance in the chamber was measured over a period of 180 minutes and divided by Area(chamber) to determine the average flux. After the rat was humanely killed, the tissue under the chamber was collected for analysis of its hyaluronan concentration ([HA]). RESULTS: All data are given as mean +/- standard error: [table: see text]. CONCLUSIONS: In the present pilot study, no significant correlations were observed, but the number of animals in each group was small (n = 3-4). Nevertheless, the results demonstrate the ability of the chamber technique to determine transperitoneal transport of water, small solutes, and protein, and to link those values directly to the structure of the tissue lying below the chamber. Thus, chronic treatment can be directly correlated with peritoneal structure and transport function. PMID- 11887842 TI - The cost barrier to peritoneal dialysis in the developing world--an Asian perspective. AB - Countries in Asia vary significantly in culture and socioeconomic status. Dialysis costs and reimbursement structures are significant factors in decisions about the rates and modalities of renal replacement therapy. From our survey of Asian nephrologists conducted in 2001, a number of observations can be made. In many developing countries, the annual cost of continuous ambulatory peritoneal dialysis (CAPD) is greater than the per-capita gross national income (GNI). The median cost of a 2-L bag of peritoneal dialysis (PD) fluid is around US$5. The absolute cost of PD fluid among countries with significant differences in per capita GNI actually varies very little. Thus, most renal failure patients can be expected to have problems accessing PD therapy in developing countries in Asia. In countries with unequal reimbursement policies for PD versus hemodialysis, a lack of incentive to prescribe PD also exists. Automated PD is nearly non existent in many developing countries in Asia. Some possible ways to reduce the cost barriers to PD in those countries include individual governments providing more public funding for treating dialysis patients; dialysate-producing companies reducing the cost of their products; physicians using appropriately smaller exchange volumes (3 x 2 L) in some Asian patients with smaller body sizes and with residual renal function; and reducing the complication rate for PD (for example, peritonitis) thereby reducing the costs required for treatment and hospitalization. PMID- 11887850 TI - Increased peritoneal membrane permeability is associated with abnormal peritoneal surface layer. AB - OBJECTIVES: We recently showed that the peritoneal surface layer may be an important barrier in modulating peritoneal membrane permeability. In the present study, we investigated the relationship between an increased peritoneal transport rate and the peritoneal surface layer. METHODS: Male Sprague-Dawley rats (n = 8) received intraperitoneal injections of 4.25% glucose dialysate daily for 1 week. Forty-eight hours after the last injection, a 4-hour dwell study using 25 mL 4.25% glucose dialysate was performed in each rat. The results were compared with those from control rats that received no intraperitoneal injections (n = 8). The peritoneal fluid and small-solute transport characteristics were evaluated. The peritoneal surface layer was studied using an electron microscope. The phospholipids content of the dialysate was also evaluated. RESULTS: Peritoneal fluid removal was significantly reduced in the daily injection group (30.6 +/- 1.3 mL) as compared with the control group (38.2 +/- 0.6 mL). The peritoneal fluid absorption rate and small-solute transport rate were also significantly higher in the daily injection group as compared with the control group. The amounts of phospholipids in the dialysate were significantly lower in the daily injection group--especially the quantity of phosphatidylcholine. However, lysophosphatidylcholine increased significantly in the daily injection group. Electron microscopy showed that the peritoneal surface layer was almost completely gone in the daily injection group, but that a dense and thick (average 4 microm) peritoneal surface layer was present on the top of the mesothelial cells in the control group. CONCLUSIONS: Our results suggest that daily injection of hypertonic glucose dialysate significantly increased the peritoneal transport rate. The increased peritoneal transport rate was associated with a significant reduction in the peritoneal surface layer and the phospholipids content of the dialysis effluent. PMID- 11887851 TI - New animal models for encapsulating peritoneal sclerosis--role of acidic solution. AB - OBJECTIVE: Encapsulating peritoneal sclerosis (EPS), in which all or part of the intestine is enveloped in a fibrous ball resembling a cocoon, is a serious complication of peritoneal dialysis (PD). The aim of the present study was to investigate whether pH-neutral or acidic dialysis solutions induce peritoneal fibrosis. DESIGN: We divided 18 male Wistar-Kyoto (WKY) rats into three groups and dialyzed them with various solutions as follows: group I, 10 mL acidic dialysis solution (pH 3.8, containing 1.35% glucose), n = 6; group II, 10 mL pH 5.0 dialysis solution, n = 6; and group III, 10 mL neutral dialysis solution (pH 7.0), n = 6. Peritoneal catheters were inserted, and dialysis solution was injected every day for 40 days. At the end of the experiment, a peritoneal equilibration test (PET) was performed. Expression of mRNA of aquaporins 1 and 4 (AQP-1 and AQP-4) in the peritoneum were studied by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: In rats treated with pH 3.8 dialysis solution, necropsy findings revealed features identical to those of EPS. The typical appearance was of granulation tissue or fibrotic tissue (or both) covering multiple surfaces. Multiple adhesions were present. In microscopic examinations, peritoneal fibrosis and loss of mesothelium were found. In rats treated with pH 7.0 dialysis solution, no signs of EPS were seen. In rats treated with pH 5.0 dialysis solution, milder changes (subserosal thickening and partial adhesion of the peritonea) were observed. The mRNA of AQP-1 and AQP-4 were expressed in the peritonea of the rats. The expression of the AQPs was significantly suppressed in rats treated with pH 3.8 dialysis solution. CONCLUSIONS: In rats, long-term intraperitoneal injection of acidic dialysis solution produced features typical of EPS in humans. Newly developed neutral dialysis solutions protected the against the development of EPS during peritoneal dialysis in rats. PMID- 11887852 TI - High glucose dialysis solutions increase synthesis of vascular endothelial growth factors by peritoneal vascular endothelial cells. AB - OBJECTIVE: Increased peritoneal vasculature has been reported in long-term peritoneal dialysis (PD), and vascular endothelial growth factors (VEGFs) have been found in dialysate. High concentrations of glucose or lactate, glucose degradation products (GDPs), and low pH of dialysis solutions are all possible factors in increased peritoneal VEGF synthesis. In this study, we investigated the effects of high glucose dialysis solutions on VEGF synthesis by peritoneal vascular endothelial cells (PVECs). METHODS: The PVECs were isolated from rat omentum and were incubated for 4 hours in three different culture media [M199 media (control), conventional dialysis solutions containing 4.25% glucose diluted with an equal volume of M199 media (HGD), and M199 media containing 118 mmol/L mannitol as an osmolar control (mannitol)]. Levels of VEGF protein in the culture supernatant were measured by ELISA, and mRNA expression was determined by Northern blot analysis. Data are presented as percent of control. RESULTS: After incubation for 4 hours, the number of cells did not differ between the 3 groups. Levels of VEGF in culture supernatant were significantly higher in the HGD group (124% +/- 19%, p = 0.006) as compared with the control and mannitol (85% +/- 10%) groups. The mRNA expression of VEGF appeared to be higher in the HGD group (128% +/- 49%) than in the control and mannitol (94% +/- 18%) groups. CONCLUSION: High glucose dialysis solutions increased VEGF synthesis by PVECs. The relationship between VEGF synthesis by PVECs and neovascularization of the peritoneum observed in long-term peritoneal dialysis patients has to be studied further. PMID- 11887853 TI - Non anticoagulant effects of heparin: implications for animal models of peritoneal dialysis. AB - Heparin is a glycosaminoglycan with well-known anticoagulant activity. That property is used in animal models of peritoneal dialysis to maintain catheter patency and to prevent the development of peritoneal adhesions. However, heparin has a host of biologic actions beyond its role as an anticoagulant. Heparin modulates the activity of various inflammatory cells, affects the synthesis of extracellular matrix, has antiproliferative effects on several cell types, and influences neoangiogenesis. By virtue of those actions, intraperitoneally administered heparin may interfere with peritoneal membrane homeostasis. The potential side effects of heparin use in animal models of peritoneal dialysis should be recognized to permit correct interpretation of experimental studies conducted in those models. PMID- 11887854 TI - Effect in a rat model of heparinized peritoneal dialysis catheters on bacterial colonization and the healing of the exit site. AB - We performed a prospective, double-blind, randomized study to evaluate whether stable surface heparinization of silicone peritoneal dialysis (PD) catheters prevents bacterial colonization or biofilm formation and improves healing of the exit site. Heparinized catheters were implanted in 20 Sprague-Dawley rats (group H) and non heparinized catheters in another 20 (group C). The PD catheters, constructed of silicon tubing with two polyester cuffs, were patterned after the standard Tenckhoff catheter. A covalent multipoint method of attachment onto polymeric surfaces was used for stable, permanent chemical immobilization of heparin on the PD catheter. Dialysis exchanges (25-mL instillation volume) were performed twice daily for 4 weeks through the permanent catheter. Prophylactic antibiotics were not used. The exit sites were evaluated at 2-week intervals. The extent of biofilm coverage on the intraperitoneal portion of the catheter (obtained at the end of the experiment) was assessed, and sonicated fluid from the catheter tip was cultured for evaluating bacterial colonization of the catheter. Exit-site scores in group H were lower than in group C (p = 0.052) at the end of week 4. Bacterial colonization tended to be less common in group H [2 of 12 catheters (17%)] than in group C [8 of 15 catheters (53%); p = 0.058], but the extent of biofilm, the peritonitis rate, and the inflammation score of tissue adjacent to the cuff were not different between the groups. Those data suggest that heparinized PD catheters can be a practical approach to the prevention of bacterial colonization and can improve healing of the exit site. PMID- 11887855 TI - The effect of icodextrin-based solutions on peritoneal transport in rats undergoing chronic peritoneal dialysis. AB - BACKGROUND: We evaluated the effect of icodextrin on peritoneal permeability and inflammation in an experimental chronic peritoneal dialysis (PD) model with repeated dwell studies (DSs) in non uremic rats. METHODS: Male Wistar rats with implanted peritoneal catheters were infused twice daily for 3 weeks with 20 mL Dianeal 3.86% (Baxter Healthcare Corporation, Deerfield, IL, U.S.A.) (n = 11) or icodextrin 7.5% (n = 12). After 10 days (DS1) and 21 days (DS2), a 4-hour DS using 30 mL icodextrin solution was performed in conscious animals. Radioiodinated serum albumin (RISA) was used as a macromolecular volume marker. Blood samples were drawn before the start of the dwell and at its end. RESULTS: We observed a steady increase in intraperitoneal volume (IPV) versus dwell time (0-240 minutes) during DS1 and DS2 in both groups. No significant differences in peritoneal permeability to solutes were observed between the groups. However, in both groups, IPV volume was significantly higher during DS2 after the 4-hour dwell time [IPV icodextrin: 34.4 +/- 1.4 mL (DS1), 35.4 +/- 1.1 mL (DS2), p < 0.002; IPV Dianeal: 34.2 +/- 0.9 mL (DS1), 35.2 +/- 0.7 mL (DS2), p < 0.01]. CONCLUSION: Changes of peritoneal permeability seen during in vivo experimental models of chronic peritoneal dialysis in rats with repeated dwell studies are comparable to results obtained in humans on continuous ambulatory peritoneal dialysis (CAPD). PMID- 11887860 TI - 4th International Meeting on Animal Models in Peritoneal Dialysis--discussion forum. PMID- 11887856 TI - Comparison of the biocompatibility of phosphate-buffered saline alone, phosphate buffered saline supplemented with glucose, and dianeal 3.86%. AB - OBJECTIVE: We compared the effects of intraperitoneal infusion of phosphate buffered saline (PBS, pH 7.4), of PBS supplemented with 3.86% glucose (G), and of standard dialysis solution [Dianeal 3.86%: Baxter Healthcare Corporation, Deerfield, IL, U.S.A. (D)] on intraperitoneal inflammation in dialyzed rats. METHODS: After catheter implantation, rats were infused on day 1 with PBS, on day 3 with PBS+G, on day 5 with D, and on day 7 again with PBS (PBS-2). After a 4 hour dwell, dialysate samples were collected and analyzed. RESULTS: All dialysate parameters studied [dialysate cell count, neutrophil:macrophage ratio (Ne:Ma), and total protein], except tumor necrosis factor alpha (TNFalpha), were comparable during both PBS infusions. During dialysis with PBS+G, the inflammatory response was suppressed as compared with the first dialysis with PBS (cell count, p < 0.001; Ne:Ma, p < 0.05; TNFalpha, p < 0.001; total protein, p < 0.001). During dialysis with D, peritoneal inflammatory parameters were further suppressed (cell count, p < 0.001 vs PBS and p < 0.01 vs PBS+G; Ne:Ma, p < 0.001 vs PBS and p < 0.05 vs PBS+G; TNFalpha, p < 0.001 vs PBS and p < 0.001 vs PBS+G; total protein, p < 0.001 vs PBS and p < 0.01 vs PBS+G). CONCLUSIONS: Hypertonicity of the dialysis fluid suppresses intraperitoneal inflammatory parameters in rats. The suppression was even more severe when Dianeal 3.86% was used. That finding could be due to the low pH and presence of GDPs in the fluid. PMID- 11887858 TI - A chronic inflammatory infusion model of peritoneal dialysis in rats. AB - OBJECTIVES: Peritoneal membrane changes are related to daily exposure to non physiologic dialysate and recurrent acute inflammation. We modified a daily infusion and inflammation model and evaluated it for fibrotic and angiogenic features. The feasibility of adenovirus-mediated gene transfer in the model was also assessed. METHODS: Peritoneal catheters were implanted in rats. Over a period of 4 weeks, the animals received a daily infusion of Dianeal 4.25% (Baxter Healthcare Corporation, Deerfield, IL, U.S.A.) with an initial three doses of lipopolysaccharide (LPS) or physiologic saline. Peritoneal fluid was assayed for transforming growth factor beta (TGFbeta) and vascular endothelial growth factor (VEGF). Animals were humanely killed at week 5. Net ultrafiltration was then measured, and tissue samples were immunostained for factor VIII. Mesenteric tissue was assayed for hydroxyproline content. Adenovirus-mediated gene transfer of beta-galactosidase was assayed by intraperitoneal administration of the virus, 4 days before the end of the experiment. RESULTS: Animals treated with either Dianeal or physiologic saline showed peritoneal membrane thickening and increased vascularity. Fibrosis was demonstrated by increased hydroxyproline concentration. Ultrafiltration was impaired. We found increased concentrations of VEGF and TGFbeta in the peritoneal fluid of animals treated with LPS and daily infusion. Adenovirus-mediated gene transfer to the peritoneal membrane was demonstrated in the model. CONCLUSIONS: Exposure to LPS and daily Dianeal or physiologic saline leads to peritoneal fibrosis and neoangiogenesis. Vascularization and glucose transport correlate with ultrafiltration failure. The present animal model mimics changes seen in humans on peritoneal dialysis and may be valuable for evaluating short-term interventions to prevent membrane damage. PMID- 11887857 TI - Replacement of glucose with N-acetylglucosamine in peritoneal dialysis fluid experimental study in rats. AB - BACKGROUND: Glucose is still used as an osmotic solute in peritoneal dialysis fluids, despite evidence of its local (peritoneal) and systemic toxicities. However a constant search is underway for a new, more biocompatible osmotic solute for peritoneal dialysis fluids. OBJECTIVE: The present study evaluated N acetylglucosamine (NAG) in a concentration of 220 mmol/L as an alternative to glucose for the osmotic solute in peritoneal dialysis fluid, during chronic peritoneal dialysis in rats. METHODS: For 8 weeks, male Wistar rats were infused with glucose-based or NAG-based dialysis fluid. Intraperitoneal inflammation and peritoneal permeability and morphology were evaluated in all rats during the study. RESULTS: Repeated intraperitoneal infusion of the NAG-based dialysis fluid resulted in a weaker intra-abdominal inflammatory reaction as compared with the reaction in rats infused with glucose-based dialysis solution. At the end of the study, the concentration of hyaluronan in the peritoneal interstitium obtained from NAG-treated rats was higher than that found in the interstitium taken from animals exposed to dialysis fluid containing glucose. Also, peritoneal permeability to total protein was lower in NAG-treated rats. CONCLUSION: As an alternative to glucose, NAG used for the osmotic solute in peritoneal dialysis solution decreases the intraperitoneal inflammatory reaction induced by the process of peritoneal dialysis and, indirectly (owing to the increased hyaluronan content in the peritoneal interstitium), diminishes peritoneal permeability to protein. PMID- 11887859 TI - Accumulation of omental mast cells during peritoneal dialysis. AB - OBJECTIVE: New vessel formation has been reported in various tissues during peritoneal dialysis (PD). In that line, mast cells can produce factors such as tryptase, chymase, or basic fibroblast growth factor that might contribute to the formation of new vessels. In the present study, the association of mast cells with neovascularization during PD was investigated. METHODS: Rats received daily 10 mL infusions of conventional 3.86% glucose-containing PD fluid over a 10-week period. The infusions were delivered through a subcutaneously implanted mini access port that was connected by catheter to the peritoneal cavity. Untreated rats served as a control group. The number of blood vessels and of mast cells in the omentum were counted. Also, the number of peritoneal mast cells was determined. RESULTS: Chronic exposure to PD fluid resulted in an increased number of mast cells in the omentum. However, no clear correlation was found between the elevated number of omental blood vessels and the number of mast cells in the omentum or in the peritoneal cavity. CONCLUSIONS: Omental mast cells accumulated dramatically upon exposure to PD fluid. The actual role of accumulated omental mast cells in the induction of angiogenesis during PD should, however, be further investigated. PMID- 11887861 TI - Adequacy targets of peritoneal dialysis in the Asian population. AB - End-stage renal disease affects a large number of patients in Asia. The percentage of patients utilizing PD varies significantly in Asian countries. Continuous ambulatory peritoneal dialysis (CAPD) accounts for about 80% of the dialysis population in Hong Kong. In this review, we address several questions related to adequacy targets in Asians: Are Asians different? Is dialysis adequacy important for Asians? What is the magnitude of the benefit and the optimal dose of dialysis prescription? Is the adequacy target realistic? The current international recommendations, including the Dialysis Outcomes Quality Initiative guidelines, are compared with some of our own data for Asian patients. Our published data on dialysis adequacy, nutrition, residual renal function, and peritoneal membrane transport showed that those factors have a significant impact on the morbidity and mortality of PD patients in Hong Kong. Our results show that solute clearance as measured by Kt/V has a significant impact on the outcome of Asian CAPD patients. Although Chinese PD patients have excellent medium-term patient and technique survival despite an apparently lower Kt/V as compared with the CANUSA standard, that favorable outcome should not prevent nephrologists from providing adequate dialysis to Asian patients. From our own data and analysis, we propose a target Kt/V of 1.9 in Asian CAPD populations. Small-volume dialysis (6 L daily) may be an acceptable compromise in some Asian populations with a smaller body size, especially with residual renal function, given the financial constraints in some developing countries. Dialysis adequacy means more than a Kt/V value; other clinical parameters are equally if not more important. Thus we should also aim at achieving adequate fluid removal and volume homeostasis, blood pressure control, good nutrition, normal acid-base balance, normal mineral metabolism, minimal anemia, and normal lipid metabolism. PMID- 11887864 TI - Mesothelial paracrystalline inclusions in continuous ambulatory peritoneal dialysis patients. AB - OBJECTIVE: Uremia is known to be followed by changes in the serous membranes of pleura, pericardium, and peritoneum. During continuous ambulatory peritoneal dialysis (CAPD), the peritoneum is exposed to altered body conditions as well as to the influence of dialysate. The aim of the present study was to examine the ultrastructure of the mesothelial cells in CAPD patients, and to compare the findings with those from studies of the peritoneum in uremic controls. Paracrystalline intracytoplasmic inclusions in mesothelial cells were objects of special interest. METHODS: Biopsies of human parietal peritoneum were studied. These were taken from 12 uremic patients during catheter implantation before the start of CAPD, and from 7 CAPD patients during catheter removal for infection or malfunction. The samples were prepared in the standard way to be studied by transmission electron microscopy (TEM). RESULTS: Paracrystalline intracytoplasmic inclusions were seen in mesothelial cells only by TEM. They appear as filamentous structures at the outer part of the inclusions, and as pearl-like structures at the core of the inclusions. Sacculate dilatations of rough endoplasmic reticulum cisternae with partly destroyed membranes and only few ribosomes were also seen, with and without densely osmiophilic filaments within the cisternae. We have found paracrystalline intracytoplasmic inclusions in mesothelial cells from uremic and CAPD patients both. According to the literature, these changes are present in one third of biopsies from uremic patients. Until now, however, they have not been mentioned in CAPD patients. PMID- 11887862 TI - Emodin ameliorates glucose-induced morphologic abnormalities and synthesis of transforming growth factor beta1 and fibronectin by human peritoneal mesothelial cells. AB - OBJECTIVE: Excessive synthesis and deposition of matrix proteins by peritoneal mesothelial cells can lead to structural and functional changes in the peritoneal membrane, jeopardizing the long-term efficacy of peritoneal dialysis (PD). Prolonged exposure to high glucose concentrations in PD fluid has been implicated as a major stimulus to matrix accumulation, through the induction of transforming growth factor beta1 (TGFbeta1). This study investigated the effect of emodin (3 methyl-1,6,8-trihydroxyanthraquinone) on TGFbeta1 and fibronectin (FN) synthesis in human peritoneal mesothelial cells (HPMCs) under high glucose concentration. DESIGN: The HPMCs were preconditioned in either 5 mmol/L or 30 mmol/L D-glucose for 2 weeks prior to the addition of emodin. Cell viability was assessed by MTT assay and lactate dehydrogenase (LDH) release. Morphology of HPMCs was studied by phase-contrast microscopy. Modulation of TGFbeta1 and FN synthesis at transcription and translation were investigated by reverse transcriptase polymerase chain reaction (RT-PCR), ELISA, and Western blot analysis. RESULTS: When cultured under 30 mmol/L D-glucose, HPMCs demonstrated increased cell volume, multinucleation, and denudation of the monolayer, as compared with cells cultured under a physiologic (5 mmol/L) glucose concentration. High glucose concentration induced TGFbeta1 synthesis by HPMCs (217.17 +/- 14.88 pg/mL at 5 mmol/L D-glucose vs 370.33 +/- 20.67 pg/mL at 30 mmol/L D-glucose, p < 0.0001), and FN synthesis was induced at transcription and translation. Mannitol at 30 mmol/L did not affect HPMC morphology; matrix synthesis was also unaltered. Administration of emodin together with 30 mmol/L D-glucose resulted in amelioration of cell enlargement and exfoliation, and abrogation of TGFbeta1 induction (370.33 +/- 20.67 pg/mL for 30 mmol/L D-glucose alone vs 260.50 +/- 17.89 pg/mL for 30 mmol/L D-glucose + emodin, p < 0.0001). Synthesis of FN induced by high glucose was also reduced by 40% in the presence of emodin. CONCLUSIONS: These findings provide the first evidence that emodin can ameliorate high glucose-induced matrix synthesis in HPMCs by suppression of TGFbeta1. Emodin may thus be useful in preserving peritoneal integrity in PD. PMID- 11887865 TI - Diagnosis and management of encapsulating peritoneal sclerosis. AB - Encapsulating peritoneal sclerosis is a rare but clinically important complication of peritoneal dialysis. In patients on long-term peritoneal dialysis, the high prevalence of the complication warrants a high index of suspicion and a low threshold for investigation--particularly in patients with ultrafiltration loss. Laparoscopic inspection of the peritoneum and peritoneal biopsy are required for definitive diagnosis. However, initial radiologic investigations may be useful. Although treatment options are based on a small number of anecdotal reports, once the diagnosis is confirmed and infection is excluded, it is reasonable for the peritoneal catheter to be removed and immunosuppressive therapy (with or without colchicine) to be commenced. Depending on the degree of bowel dysfunction at diagnosis, total parenteral nutrition may be commenced. In the absence of improvement, surgical intervention with total intestinal enterolysis may be attempted, recognizing the high morbidity and mortality associated with operative procedures in these patients. The ultimate aim is to prevent the development of encapsulating peritoneal sclerosis. Although a link is not proven, the development of more biocompatible dialysis fluids with less potential to result in glycosylation of the peritoneal membrane proteins will, it is hoped, reduce the occurrence of this devastating complication of peritoneal dialysis. PMID- 11887867 TI - The greater incidence of encapsulating peritoneal sclerosis is not the result of overdiagnosis. PMID- 11887863 TI - Aspirin protected the nitric oxide/cyclic GMP generating system in human peritoneum. AB - OBJECTIVE: Changes in the expression of endothelial nitric oxide synthase (eNOS) in the peritoneum could be involved in the peritoneal dysfunction associated with peritoneal inflammation. The aim of the present study was to analyze the effect of Escherichia coli lipopolysaccharide (LPS) on eNOS expression in samples of human peritoneum. The effect of aspirin, a drug with anti-inflammatory properties, was also determined. RESULTS: The eNOS protein expressed in human peritoneal tissue was reduced by LPS (10 microg/mL) in a time-dependent manner. The eNOS was expressed mainly in capillary endothelial cells and mesothelial cells. Anti-inflammatory doses of aspirin (1-10 mmol/L) restored eNOS expression in LPS-stimulated human peritoneal tissue samples. The main intracellular receptor of NO, soluble guanylate cyclase (sGC), was also downregulated by LPS. This effect was prevented by aspirin (5 mmol/L). CONCLUSION: Protein expression of the eNOS-sGC system in the peritoneal tissue was downregulated by LPS. High doses of aspirin protected both eNOS protein expression and sGC in human peritoneum. These findings suggest a new mechanism of action of aspirin that could be involved in the prevention of peritoneal dysfunction during inflammation. PMID- 11887866 TI - Encapsulating peritoneal sclerosis in Japan: prospective multicenter controlled study. AB - OBJECTIVE: Encapsulating peritoneal sclerosis (EPS) is recognized as a serious complication of peritoneal dialysis. The aim of this study was to determine the incidence, clinical features, and variation in mortality rates for EPS. DESIGN: A prospective multicenter design was used, in which peritoneal dialysis patients were pre-registered by facilities across Japan and the incidence of EPS was observed in the registrants. The registrants were followed for a total of 4 years to accurately observe the onset of EPS. RESULTS: As of April 1999, 2216 peritoneal dialysis patients from 64 facilities were registered. By the end of March 2001, 332 patients had dropped out, and 17 of the dropouts had developed SEP. The incidence was 0.77%. After excluding 110 patients who died, the incidence in 2106 patients was 0.81%. The incidence of EPS increased with the duration of peritoneal dialysis. Of the 17 patients with EPS, 12 developed the condition after discontinuing peritoneal dialysis and changing to hemodialysis. During the 2-year survey period, 6 of the 17 EPS patients died. The interval from onset to death was 10.8 +/- 5.8 months (range: 3-19.5 months). CONCLUSIONS: From this prospective multicenter study, the current incidence of EPS is 0.77% (0.81% when dropout owing to death is censored). After a follow-up of 2 years, we conjecture that the incidence of EPS will increase. The incidence, etiology, and prognosis of EPS will be further clarified by periodic observation of dropouts until the end of March 2003. PMID- 11887869 TI - Clinical outcome in continuous ambulatory peritoneal dialysis patients is not influenced by high peritoneal transport status. AB - OBJECTIVES: We undertook this study to examine the influence of demographics, peritoneal membrane transport characteristics, nutrition indices, dialysis adequacy, and comorbid diseases on patient survival on continuous ambulatory peritoneal dialysis (CAPD), and to identify whether high peritoneal membrane transport is an independent risk factor for mortality on CAPD. DESIGN: Our retrospective study was carried out in CAPD patients in a large tertiary care teaching hospital. METHODS: Until December 2000, we followed 212 patients who started CAPD between 1994 and 1997 and who underwent a peritoneal equilibration test (PET) within 3 months of CAPD initiation. RESULTS: By univariate analysis, comorbid diseases, old age, high peritoneal transport, and serum albumin predicted patient mortality. Independent predictors of mortality as determined by the Cox proportional hazard model included diabetes mellitus, cardiovascular disease, serum albumin, and old age. High peritoneal transport failed to independently influence mortality in our CAPD patients. PMID- 11887870 TI - Peritoneal dialysis solutions. AB - After several decades of experience of peritoneal dialysis therapy, we now understand the peritoneal membrane and the causes of its changes during long-term dialysis much better. Several new, more biocompatible solutions are available in clinic today, and the outcome of peritoneal dialysis therapy is expected to be further improved. However, limitations with the currently available peritoneal dialysis solutions still exist, and continual efforts are needed to develop solutions that are more efficient and more membrane-friendly. With the better understanding of the role of fluid balance in peritoneal dialysis, we believe that development of peritoneal dialysis fluid that protects the peritoneal surface layer (and thus the integrity of the peritoneum, thereby improving peritoneal fluid removal) may be an area of research in the near future. PMID- 11887868 TI - The problem of the high transporter: is survival decreased? AB - The mechanisms responsible for the problem status of high transporters are diverse. High transporters have increased protein losses that may play a role in the presence of hypoalbuminemia and malnutrition. On the other hand, high transport induces increased glucose absorption, which may in turn be responsible for anorexia and increased atherogenesis--issues not discussed here, but clearly of major importance. And finally, the impaired ultrafiltration present in the high transporter leads to fluid overload, which is one of the driving forces for ventricular hypertrophy, hypertension, and increased cardiovascular risk- cardiovascular events being most prevalent cause of death in dialysis patients. All of the factors previously discussed--and others--may preclude some high transporters from being good candidates for peritoneal dialysis. Yet many others may still do well if the prescription is individually tailored to the particular patient. PMID- 11887871 TI - What happens to the peritoneal membrane in long-term peritoneal dialysis? PMID- 11887874 TI - Partitioning of marine antifoulants in the marine environment. AB - The partitioning behaviour of the organic biocides, Irgarol 1051 and diuron and two inorganic biocides (copper and zinc) was investigated using six sediments of differing physico-chemical properties collected from unimpacted sites along the south coast of England. The kinetics of sorption and equilibrium partitioning between the sediments and seawater were investigated over a period of 20 days. Resulting organic carbon/water partition coefficients (log Koc) were related to suspended sediment concentration and ranged from 2.28 to 5.20 for diuron; and from 2.41 to 4.89 for Irgarol 1051. Sediment/water partition coefficients (log Kp) for copper and zinc varied from 2.46 to 5.08 l/kg and from 2.49 to 4.97 l/kg, respectively. Kinetic data indicated that there were significant interactions between the dissolved and particulate phases at the start of the experiments, just after mixing. This is thought to be a result of redistribution of organic carbon between the two phases. PMID- 11887872 TI - In vitro superiority of dual-chambered peritoneal dialysis solution with possible clinical benefits. PMID- 11887873 TI - Lanthanide concentrations in freshwater plants and molluscs, related to those in surface water, pore water and sediment. A case study in The Netherlands. AB - Industrial emissions of lanthanides to aquatic ecosystems increase, but knowledge of the environmental fate of these metals is limited. Here we focus attention upon the distribution of lanthanides in freshwater ecosystems, describing lanthanide partitioning between sediment, water and biota. Since lanthanides are often used as oxidation-state analogues for actinides, their distribution can reflect long-term behaviour of the radioactive transuranics. Concentrations of all 14 naturally occurring lanthanides were measured by ICP-MS in Sago pondweed (Potamogeton pectinatus), common duckweed (Lemna minor), seven different mollusc species (tissue and shell), two sediment fractions (< 2 mm and < 63 microm), surface water and sediment pore water from five locations in The Netherlands. In all samples, the typical 'saw-tooth' lanthanide pattern was observed, which implies that lanthanides are transported as a coherent group through aquatic ecosystems. Typical deviations from this pattern were found for Ce and Eu and could be explained by their redox chemistry. The variation in concentrations in abiotic fractions was limited, i.e. within one order of magnitude. However, variations of up to three orders of magnitude were observed in biotic samples, suggesting different affinities among organisms for lanthanides as a group, with significant differences only among molluscs and pondweed samples in relation to sampling location. For P. pectinatus it was shown that pore water was the most important lanthanide source, and for snails, food (plants) seems to be the dominant lanthanide source. Lanthanides were not equally distributed between mollusc shell and tissue and the ratio of lanthanide concentrations in shell and tissue were dependent on the sampling location. Shells contained much lower concentrations and were relatively enriched in Eu, and to a lesser extent in Ce. Bioconcentration factors for lanthanides in plants and snails relative to surface water were typically between 10000 and 100000 l x kg(-1) dry matter, while sediment-water partition coefficients were between 100000 and 3000000 l x kg(-1) dry matter. There was a low extent of biomagnification in the plant-to-snail system, with a maximum biomagnification factor of 5.5. Many distribution coefficients displayed a slight decrease with atomic number. This can be attributed to the general increase in ligand stability constants with atomic number, keeping the heavier lanthanides preferentially in solution. PMID- 11887875 TI - Ion chromatographic separation and determination of phosphate and arsenate in water and hair. AB - A simple and sensitive method for the sequential determination of phosphate and arsenate was developed based on initial ion chromatographic separation followed by detection as the ion-association complex formed by heteropolymolybdophosphate and arsenate with bismuth. With 200 microl sample injection and separation on a AS4A-SC column using an eluent of 3.5 mM sodium hydrogen carbonate-10.0 mM sodium hydroxide, the detection limits which are calculated as the concentration equivalent to twice the baseline noise, were found to be 0.8 microg/l and 4.2 microg/l for P and As, respectively. Spiked samples were analyzed and recoveries were found to be satisfactory in the range of 95-105% for phosphate and 90-105% for arsenate. Samples of water and hair were analyzed by the proposed method. PMID- 11887877 TI - Predicting the outcome of sciatica at short-term follow-up. AB - BACKGROUND: The prognostic value of the clinical findings elicited in the patient presenting with sciatica is unknown. AIM: To investigate whether history and physical examination findings can predict outcome. DESIGN OF STUDY: Prospective study of prognostic factors. SETTING: A sample of primary care patients with sciatica. METHOD: Short-term favourable outcome was registered as improvement perceived by the patient after two weeks. Long-term failure was defined as eventual surgery or lack of improvement after three months. RESULTS: The signs and symptoms that most consistently predicted an unfavourable outcome were: a disease duration of more than 30 days; increased pain on sitting; and more pain on coughing, sneezing or straining. The straight leg raising test and, to a lesser degree the reversed straight leg raising test, were the most consistent examination findings associated with poor outcome. Chances of short-term improvement were also related to the body weight relative to the length. CONCLUSION: The predictors in this study can indicate the prognosis of patients with sciatica at an early stage. Knowledge of these prognostic factors may help to fine tune treatment decisions and improve patient selection in trials of conservative therapy strategies. PMID- 11887876 TI - A concordance-based study of metaphoric expressions used by general practitioners and patients in consultation. AB - BACKGROUND: All languages use metaphoric expressions; some deliberately chosen, some (for example, 'digesting information') not usually perceived as metaphoric. Increasingly, it is suggested metaphoric expressions constrain the way we conceptualise the world, as well as being a means of achieving stylistic effect. AIM: To study metaphoric expressions used by doctors and patients in general practice. DESIGN OF STUDY: Concordance-based language analysis of spoken data. METHOD: A database containing transcriptions of 373 consultations with 40 doctors in a UK general practice setting was scrutinised for metaphoric expressions, using 'concordancing' software. Concordancing enables identification of strings of text with similar lexical properties. Comparators (for example, 'like'), selected verb-types (for example, of feeling), and the verb 'to be' were used as starting points for systematically exploring the data. Quantitative and qualitative thematic methods were used in analysis. RESULTS: Doctors and patients use different metaphors. Doctors use mechanical metaphors to explain disease and speak of themselves as problem-solvers' and 'controllers of disease'. Patients employ a range of vivid metaphors, but fewer metaphors of machines and problem/solution. Patients use metaphors to describe symptoms and are more likely to use metaphoric language at the interface of physical and psychological symptoms ('tension, 'stress'). CONCLUSION: The different patterns of metaphoric expression suggest that doctors make limited attempts to enter the patients' conceptual world. This may not be a bad thing. One function of the consultation may be to reinterpret vivid and unique descriptions as accounts of the familiar and systemically comprehensible. Doctors may use different conceptual metaphors as a reassuring signal of expertise. PMID- 11887878 TI - Cardiovascular risk factors and disease in general practice: results of the Nijmegen Cohort Study. AB - The relationship between risk factors and cardiovascular disease (CVD) was determined using data from the Nijmegen Cohort study, an 18-year prospective study. In 1977, cardiovascular risk factors were measured in 7092 Caucasian males and females from six general practices: age, smoking, serum cholesterol, blood pressure, body mass index and a family history of CVD were related to subsequent CVD mortality and morbidity. Most patients had more than one risk factor in particular among men. A significant relationship between risk factors and CVD was demonstrated. In men, a relative risk (RR) of 1.8 was found for both high blood pressure and smokers. A positive family history yielded a RR of 1.8. The risk increased gradually with the number of risk factors; 38% of the group with all risk factors suffered a CVD within 18 years. In women, comparable RRs were found but the low absolute risk of CVD should indicate against the treatment of cardiovascular risk factors. PMID- 11887880 TI - The crystal structure of KR-21042, an analgesic capsaicinoid. AB - The crystal structure of KR-21042, N-(3-Phenylpropyl)-4-hydroxy-3 methoxyphenylacetamide, was determined by single crystal X-ray diffraction analysis. The compound was recrystallized from a mixture of ethylacetate and n hexane in monoclinic, space group P21/c, with a = 16.622(1), b = 6.215(1), c = 15.802(1) A, P= 104.97(1), and Z = 4. The calculated density is 1.261 g/cm3. The structure was solved by the direct method and refined by full matrix least squares procedure to the final R value of 0.068 for 2332 observed reflections. PMID- 11887879 TI - Drop the anchor, not the ANCA. PMID- 11887881 TI - Deaths from chickenpox. Epidemiology of chickenpox in United Kingdom needs further investigation. PMID- 11887882 TI - Deaths from chickenpox. Healthcare workers should not be forgotten. PMID- 11887883 TI - Deaths from chickenpox. Chickenpox associated morbidity may be long term. PMID- 11887884 TI - Deaths from Chickenpox. Extracorporeal membrane oxygenation has important role. PMID- 11887885 TI - Secondary prevention of coronary heart disease. Secondary prevention programmes may reduce overall mortality in high risk patients. PMID- 11887886 TI - Secondary prevention of coronary heart disease. Improved outcomes need to be defined. PMID- 11887887 TI - Dysphagia. With endoscopic pouch stapling, pouch excision is no longer necessary. PMID- 11887890 TI - Tracking reliability for space cabin-borne equipment in development by Crow model. AB - Objective. To study and track the reliability growth of manned spaceflight cabin borne equipment in the course of its development. Method. A new technique of reliability growth estimation and prediction, which is composed of the Crow model and test data conversion (TDC) method was used. Result. The estimation and prediction value of the reliability growth conformed to its expectations. Conclusion. The method could dynamically estimate and predict the reliability of the equipment by making full use of various test information in the course of its development. It offered not only a possibility of tracking the equipment reliability growth, but also the reference for quality control in manned spaceflight cabin-borne equipment design and development process. PMID- 11887888 TI - Management of Helicobacter pylori infection. Eradication treatment can be tailored in patients undergoing endoscopy. PMID- 11887889 TI - Management of Helicobacter pylori infection. Dental plaque is a potential reservoir of Helicobacter pylori. PMID- 11887891 TI - [Changes of early components of auditory ERPs during HDT-simulated weightlessness]. AB - Objective. To study changes of early components of the auditory ERPs during simulated weightlessness. Method. The event-related potentials (ERPs) during an auditory location discrimination were compared between head down tilt (HDT) and head up tilt (HUT) conditions in 14 normal subjects. Result. The early components of T-ERPs and NT-ERPs decreased significantly during HDT as compared with that during HUT. The reduction of mean potential amplitude during HDT was more marked at left-forehead area (F5). Conclusion. These data provided further evidence showing that the early activity process of brain was also affected by simulated weightlessness. PMID- 11887892 TI - [Effect of lower body negative pressure and rotating-table simulated push-pull effect in flight on cardiovascular function]. AB - Objective. To explore the effect of push-pull effect (PPE) simulated by lower body negative pressure (LBNP) rotating-table, and observe the physiological responses to push-pull maneuver. Method. A special LBNP rotating-table was used to simulate the push-pull maneuver. 8 healthy adults participated randomly in two experiments. One was simulated PPE test, which include a series of head-up stand (HUT, +1 Gz) for 1 min, then in head-down stand (HDT, -1 Gz) for 30 s and again in HUT combining LBNP (-50 mmHg) for 10 min. The other one was control test, which only consists of HUT combining LBNP (-50 mmHg) for 10 min. Changes of heart rate (HR), blood pressure (BP), basic impedance (Z0) , stroke output (SO) , cardiac output (CO) and total peripheral resistance (TPR) were monitored by electrical impedance instrument during the experiment. Result. During simulated PPE experiment, 3 subjects presented presyncopal symptoms, with average standing time of 8.99 +/- 1.47 min, while during control test, all the subjects completed HUT combining LBNP for 10 min. In simulated PPE experiment, as compared with HUT (control), HR, Z0 during HDT were significantly lowered, while SV and CO were increased significantly. During HUT + LBNP, HR, Z0 and TPR were significantly higher, while SV and CO were significantly lower than that of control and HDT. SBP was increased significantly than control value when "HUT + LBNP" started, but during the whole process of "HUT + LBNP ", it became significantly lowered. In control experiment, the above mentioned indexes showed the same trend of as change compared with the control, however, the percentage of the change was lower than simulated PPE test, the change percentage in HR was not including. Conclusion. After headstand, head-up stand combining LBNP caused cardiovascular function descends, the degree was larger than simple head-up stand combining LBNP. LBNP rotating-table can be used to simulated push-pull effect. PMID- 11887893 TI - [Daily 1 h standing can prevent depression of myocardial contractility in simulated weightless rats]. AB - Objective. To observe and compare the effect of daily standing of rats for different durations in alleviating the depression of myocardial contractility induced by simulated weightlessness. Method. Fifty male Sprague-Dawley rats were randomly assigned to five groups: simultaneous control (CON), four-week tail suspension (SUS), SUS plus daily 1 h standing (STD1), SUS plus daily 2 h standing (STD2) and SUS plus daily 4 h standing (STD4). Four weeks later, wet weights of the left soleus, testis and adrenal gland were measured, and the isometric contractile tension and its time parameters of isolated perfused papillary muscles from rats of various groups were examined. Result. Compared with CON, the wet weights of testis in different treatment groups decreased significantly (P<0.05). The wet weight of soleus in SUS decreased by 58.9% (P<0.01); but in STD1, STD2 and STD4, decreased by 38.5%, 24.0% and 11.0% (P<0.01, or P<0.05) respectively. The relative protections were 34.7%, 59.2% and 81.4% (P<0.01) respectively. In SUS group, developed tension (DT), peak rate of tension rise (+dT/dtmax) and peak rate of tension fall (-dT/dtmax) decreased by 32.2%, 29.2% and 30.7% (P<0.05), and time to peak rate of tension rise (TPP) and time to peak tension (TPT) prolonged by 21.2% and 11.0% (P<0.05), respectively. Whereas in STD1, STD2 and STD4, all the parameters reflecting myocardial contractility (like DT, + dT/dtmax, -dT/dtmax and TPP), did not show any significant change as compared with those of CON. However, being an exception, TPT still showed obvious prolongation (P<0.05) in STD1. Conclusion. Daily 1 h standing could prevent depression of myocardial contractility in rat induced by medium-term simulated weightlessness. However, with respect to prevention of atrophic changes in soleus muscle and testis, daily standing for 1 h, 2 h, or 4 h was only partially effective and even totally devoid of any effect, respectively. PMID- 11887894 TI - [Effects of repeated high +Gz exposure on several enzyme activities in cardiomyocytes in rats and some protective measures]. AB - Objective. To determine the changes of several myocardial enzymes in rats after repeated high +Gz exposure and the protective effects of preconditioning of low-G exposure and tea polyphenols (TP). Method. Thirty-two male Wistar rats were randomly divided into 4 groups (n = 8 each): control group (group A), +10 Gz group (group B), low-G preconditioning group (group C) and TP protection group (group D). Group B, C and D were exposed to repeated +10 Gz stress (each for 30s, 5 times/d with +1 Gz 1 min intervals, 3 d/wk, 3 weeks in total), but group A was only submitted to +1 Gz for 5 min. Group C was exposed to +2 Gz for 5 min about 1 h prior to +10 Gz stress. Additionally, TP (200 mg/kg) was given orally to group D about 1 h prior to the +Gz experiment, while distilled water was given to groups A, B and C instead. On the next day after the last centrifuge run, the hearts were taken out immediately for making frozen tissue sections. Enzyme histochemical staining and image analysis were carried out for acid phosphatase (ACP), succinate dehydrogenase (SDH), cytochromeoxidase (Cyt aa3) and adenosine triphosphatase (ATPase). Result. As compared with the control, the activities of ACP and SDH in +10 Gz stressed rats decreased significantly (P<0.05), and there was a declining trend for Cyt aa3. But, low-G preconditioning and TP had protective effects on +10 Gz stress-induced reduction of these enzymatic activities (P<0.05). Conclusion. The results showed that repeated high +Gz exposure could bring about decreases of activities in ACP (the marker enzyme of lysosome) and SDH (the marker enzyme of mitochondrial endomembrane), which indicated a reduction of oxidative metabolism in myocardial tissue; but preconditioning with low-G and natural antioxidant TP had protective effects. PMID- 11887895 TI - [Experimental study on ergonomical color matching design of virtual crew cabin layout in manned spacecraft]. AB - Objective. To approach general principles of color matching for crew module layout and to provide its ergonomical evaluation with basic data. Method. First, according to some ergonomic rules a virtual reality experimental system was set up, then 64 subjects of different ages and with some background of spaceflight were offered a color matching example according to their own choice in advance. Finally, all the hues, saturations, and lightnesses of the selected colors and their total number were statistically analyzed by SPSS 8.0 software. Result. After choosing the colors for items (standard cabinets, floor, handrails, supports and etc.) in the crew cabin, the mean kinds of color hue matching in the cockpit was 5. In addition, above half of subjects endorsed the example colors but its saturation and lightness were a little higher than those of the example every time. Although its distribution was discrete, there still was a common agreement on color matching (about 50%). Conclusion. When the color matching of crew module in long time flight was ergonomically designed, generally, cool and warm hues should be taken into consideration, and their total number need be controlled to be under 5 so as to satisfy human psychological characters. PMID- 11887896 TI - [Geometrical dummy used for ergonomic evaluation on early stage cockpit conceptual design]. AB - Objective. Basing on the need of cockpit ergonomic design, to set up a geometrical dummy. Method. Surface partition of the dummy, source of the curved face data, express of the curved face, degree of freedom of the human joint, coordinate system of the parts, position of the human segment centroid and weight of the human segment were cleared up first, then geometrical dummy was displayed with three-dimensional software. Result. The geometrical dummy could simulate some basic body movements and postures for ergonomic analysis. Conclusion. The geometrical dummy was suitable for present study and engineering design. PMID- 11887898 TI - [Effects of Rholida on the free radical metabolism and serum creatine kinase after exercise at plateau]. AB - Objective. To study the effect of Rholida on free radical metabolism and serum creatine kinase CK) after exercise at plateau. Method. After staying at high altitude (4100 m) for 20 d, 40 healthy young men were divided into 4 groups randomly (Rholida, Acetazolamide, Xi' s capsule and control, 10 men each group). And their SOD, MDA, GSH-Px CK, and CK-MB were determined respectively. Before, after taking drugs and after finishing the 5 min-stair-exercise. Result. Before taking drugs and after exercise, MAD GSH-Px, CK, CK-MB, increased as compared with quiet state (P<0.05, P<0.01), but SOD showed no significant chang (P>0.05). After taking drugs for 6 d, those who took Rholida, Acetazolamide and Xi's capsule, their MAD, GSH-Px CK, CK-MB increased after exercise as compared with quiet state (P<0.05). In Rholida group SOD increased and had significant change (P<0.05); but there was no significant change in Acetazolamide, Xi' s capsule group, SOD increased, MDA decreased (P<0.05), CK, CK-MB had no significant change (P>0.05), GSH-Px increased in Xi's group (P<0.05), but not in Acetazolamide group (P>0.05). SOD, GSH-Px increased, MDA, CK-MB decreased in Rholida group after taking drugs and the changes were significant (P<0.01). In Acetazolamide and Xi's capsule group, GSH-Px increased significantly, MDA, CK, CK-MB decreased significantly (P<0.05), but SOD didn't (P>0.05). Conclusion. Rholida, Acetazolamide, Xi's capsule could regulate the disorder of free radical metabolism at plateau and Rholida had advantage over the others. PMID- 11887897 TI - [A control study of personality characteristics and variation of plasma peptide in pilots with neurosis]. AB - Objective. To study the relationship between personality type and variation of plasma peptides in pilots with neurosis. Method. A case-control study was used. 124 male pilots were evaluated with Eysenck's personality checklist, and then level of certain plasma peptides, such as vasoactive intestinal peptide (VIP), beta-endorphin (beta-EP) and angiotensin-II (A-II) were determined. Result. There were significant difference in personality characteristics and personality types between pilots with neuroses and the control. The contents of VIP and beta-EP in plasma showed visible difference between disease group and control. Content of beta-EP in those with inner-unstable type personality was lowest among all the various types. Conclusion. Personality characteristics were different between pilots with neurosis and controls. Levels of VIP and beta-EP in disease group were lower than those in the control. Different personality types had different levels of beta-EP in pilots with neurosis. PMID- 11887899 TI - [Analysis of the changes of temporomandibular joint under repeated +Gz stress]. AB - Changes of temporomandibular joint (TMJ) under repeated +Gz stress were discussed. From the etiological point of view in TMJ, many papers in the fields of aviation medicine, microcirculation, maxillofacial surgery and bone surgery were reviewed. +Gz forces can cause inadequacy of blood of oxygen supply to TMJ area. This situation can be worsened by release of free radical agent and cellular factors, ischemia/reperfusion injury, and/or hemorrheologic changes. Furthermore, G-induced injury of cervical muscles and spine may break the maxillofacial muscle chain balance. In addition to the above factors, mental stress may do harm to TMJ. This paper introduced the researches on this area in an attempt to enlighten the concern about TMJ responses to increased +Gz acceleration forces. PMID- 11887900 TI - [Heat shock protein 70 and stimulation induced by +Gz]. AB - High sustained +Gz acceleration produced by modern high performance aircrafts can induce brain damage because of ischemia and hypoxia. Heat shock protein 70 (HSP70) is one of the proteins broadly exist in organisms. The expression of HSP70 is an important reaction in the brain, and it has close relationship with brain ischemic injury. Now, the common view thinks that the expression of HSP70 during stimulation can protect nerve cells. This paper reviewed the biologic characteristics of HSP70 and the protective effect of HSP70 on nerve cells after brain ischemia. Also, the relationship of HSP70 expression and exposure value of +Gz were discussed. PMID- 11887901 TI - [Mechanotransduction in bone]. AB - This article reviewed the progresses in the research of mechanisms of bone adaptation. The process of mechanotransduction can be divided into four distinct steps: mechanocoupling; biochemical coupling; transmission of signal; effector cell response. Through these four steps, the loads acting on the bones are transduced into biochemical signals, and then change the function of bone cells, finally induce the changes of bone structures to adapt the mechanical environment. PMID- 11887902 TI - Ethics and aged-care managers. PMID- 11887904 TI - Hope and responsibility in clinical settings: two reflections on Jewish life and death. Are scientific truths the only truths? PMID- 11887905 TI - Hope and responsibility in clinical settings: two reflections on Jewish life and death. Looking to "the world to come" and physician-assisted suicide. PMID- 11887903 TI - Designer babies. PMID- 11887907 TI - Expanding coverage to parents through Medicaid Section 1931. PMID- 11887908 TI - State approaches to expanding family coverage. PMID- 11887906 TI - The economic impacts of the tobacco settlement. AB - Recent litigation against the major tobacco companies culminated in a master settlement agreement (MSA) under which the participating companies agreed to compensate most states for Medicaid expenses. Here the terms of the settlement are outlined and its economic implications analyzed using data from Massachusetts. The financial compensation to Massachusetts (and other states) under the MSA is substantial. However, this compensation is dwarfed by the value of the health impacts induced by the settlement. Specifically, Medicaid spending will fall, but only by a modest amount. More importantly, the value of health benefits ($65 billion through 2025 in 1999 dollars) from increased longevity is an order of magnitude greater than any other impacts or payments. The net efficiency implications of the settlement turn mainly on a comparison of the value of these health benefits relative to a valuation of the foregone pleasure of smoking. To the extent that the value of the health benefits is not offset by the value of the pleasure foregone, the economic impacts of the MSA will include a share of these health benefits. PMID- 11887909 TI - ERISA complicates state efforts to improve access to individual insurance for the medically high risk. PMID- 11887910 TI - Continuation of papers presented at the Fifth International Symposium on Basic Environmental Problems of Man in Space, Washington, D.C., 27-30 November 1973. PMID- 11887911 TI - Study of water-salt metabolism and renal function in cosmonauts. AB - In manned space flights the renal function and water-salt metabolism undergo substantial changes. With the reserve capabilities of kidneys in mind, their function and regulation of the water-salt balance were investigated in cosmonauts postflight and in Earth-bound simulation experiments with the aid of water loading, hormonal injections (pituitrin, engiotensin, DOCA, ACTH); water- and ion release were also studied during LBNP and physical exercises. The cosmonauts who performed space flights of 2 to 5 days showed water retention and increased urine excretion of salts during the first postflight days in response to a water load. After the 18-day flight water excretion remained unchanged whereas salt excretion increased. The capacity for osmotic concentration and urine dilution did not alter. The study of the hormonal effect in simulation experiments of different duration demonstrated a normal renal response to the hormonal excretion. After the LBNP tests and physical exercises the water- and salt-excretion declined; a correlation between the level of water- and salt-excretion and the level of these loads was established. The data on the blood- and urine-ionic composition, excretion of nitrogen metabolites, and hormones postflight as well as the results of load and functional tests suggest that changes in the renal function of cosmonauts in weightlessness are associated with regulatory effects on the kidney rather than disturbances in the function of nephron cells. PMID- 11887912 TI - Characteristics of metabolism during prolonged water immersion. AB - The effect on the organism of a 12-day stay in a water immersion medium (head on the water surface) was studied on 10 subjects. The condition of the metabolic processes in the subjects was judged from the investigation of the parameters of protein and carbohydrate metabolism, acid-base equilibrium, the activity of a number of enzymes and steroid hormones. The venous blood and diurnal urine served as the material for conducting the corresponding biochemical determinations. The stay in an immersion medium is accompanied by a significant increase in the content of residual nitrogen in the blood, by a reduction of the creatinine content in the blood and by an increase of the creatine content of the blood with the simultaneous increase of the excretion of creatinine and creatine with the urine, by a significant increase of the glucose and lactate content in the blood, by the development of acidosis of a respiratory and metabolic character, as well as by a significant increase of the activity of creatine phosphokinase and the isoenzyme fraction of lactate dehydrogenase3. In all the subjects, an expressed intensification of the glucocorticoid and androgenic functions of the adrenal cortex is noted during the experimental period. PMID- 11887913 TI - Changes in the vestibular function during space flight. AB - An analysis of observations and investigations carried out in space flight has shown that some cosmonauts and astronauts have experienced vestibular disorders during the transition to weightlessness. Vestibular-sensory disorders include: Spatial illusions (the feelings of falling down, being in an upside-down position, the sensations of rotation of the craft or the body) and vertigo occurring during the onset of the orbital flight and head movements; Feelings, similar to those experienced in response to Coriolis accelerations on the Earth, which occasionally develop in weightlessness during the spacecraft rotation upon abrupt head and body movements and restrained feet; Feelings "of the load on the vestibular analyser which is unlike any Earth-bound effects" upon abrupt head movements during the first hours of an orbital flight and "a prolonged movement" during the switch-off of thrusters in weightlessness. Vestibular-vegetative disorders comprise a complex of symptoms similar to those of motion sickness: loss of appetite, stomach awareness (12%), hypersalination, nausea (9.6%) and vomiting (4.8%). Soviet studies suggest that the vestibular tolerance to the flight effects depends on the natural stability and training to the cumulative effect of adequate vestibular stimuli. This has been used in the development of the system of vestibular selection. Changes in the vestibular function seem to play the major role in the development of motion sickness in weightlessness, extra-labyrinthine factors being contributory. The current hypotheses have not yet been adequately confirmed in experiments. A detailed physiological analysis allows the conclusion that the decisive factor in the development of motion sickness may be the disturbance of the function of analysers responsible for spatial orientation which take the form of sensory conflicts as well as an altered reactivity of the organism due to the hemodynamic rearrangement. PMID- 11887914 TI - The control of posture and movements during REM sleep: neurophysiological and neurochemical mechanisms. AB - Mammalian sleep is characterized by synchronized sleep, in which high-amplitude, low-frequency waves appear in the electroencephalogram, and desynchronized sleep, characterized by small-amplitude, high-frequency waves, absence of tonic muscle activity, and rapid eye movements (REM), which are associated in humans with dreams. The postural atonia typical of desynchronized sleep is due to postsynaptic inhibition of spinal motoneurons resulting from tonic activation of a bulbospinal inhibitory system. Superimposed on this background of postural atonia, a motor pattern appears, characterized by rapid contractions of the limb musculature synchronous with the REM bursts. Simultaneously one observes phasic inhibition of transmission of somatic afferent volleys to motoneurons and ascending spinal pathways. The bursts of REM depend upon rhythmic discharges of vestibulo-oculomotor neurons, due to extralabyrinthine volleys originating from the brainstem. Ascending and descending vestibular volleys are also able to excite corticospinal and other supraspinal descending neurons responsible for the motor events synchronous with the bursts of REM. Activation of cholinergic neurons located in the brainstem reticular formation reproduces the postural atonia typical of desynchronized sleep, as well as the phasic events characterized by the REM bursts and the related changes in spinal cord activities. Even in this instance the bursts of REM and the related spinal effects depend upon rhythmic changes in the discharge of vestibular nuclear neurons. Experimental evidence indicates that the cholinergic reticular neurons fire asynchronously, thus being able to trigger the bulbospinal inhibitory system responsible for postural atonia. Even the vestibulo-oculomotor neurons are activated by these cholinergic reticular neurons; however, the continuous stream of these extralabyrinthine impulses is transformed into rhythmic changes of discharge of the vestibular nuclear neurons due to the presence of inhibitory neurons interposed with the vestibulo-oculomotor system. Waxing and waning in the activity of these cholinergic reticular neurons accounts for the regular occurrence of the cholinergically induced bursts of REM. PMID- 11887915 TI - Effects of muscle electrostimulation during simulated weightlessness. AB - In a 45-day experiment test subjects were exposed to bed rest with their heads down at -4 degrees C. Twice a day their muscles of the stomach, back, femur, and shin were stimulated with electric current for 25-30 min. The value of muscle tension was close to their maximum voluntary contraction. The main objective was to prevent muscle atrophy and to maintain their trophic and functional state. Physiological measurements were carried out together with morphological, cytochemical, and biometric evaluations. The tissue removed during biopsy from M. soleus 7 days before the test and on the 30th hypokinetic day was used as substrate. Electrostimulation affected favourably the tone and strength of muscles as well as their static and dynamic endurance. Morphological studies showed a positive effect of electrostimulation on the muscle tissue, preventing the development of atrophic processes. During the first post-hypokinetic day orthostatic tolerance increased. PMID- 11887916 TI - The biological effectiveness of HZE-particles of cosmic radiation studied in the Apollo 16 and 17 Biostack experiments. AB - The Biostack experiments I and II were flown on board the Apollo 16 and 17 command modules in order to obtain information on the biological damage produced by the bombardment of heavy high-energy (HZE) particles of cosmic radiation during spaceflight. Such data are required for estimating radiation hazards in manned spaceflight. Seven biological systems in resting state (Bacillus subtilis spores, Colpoda cucullus cysts, Arabidopsis thaliana seeds, and eggs of Artemia salina, Tribolium castaneum and of Carausius morosus) were accommodated in the two Biostacks. By using a special sandwich construction of visual track detectors and layers of biological objects, identification of each hit biological object was achieved and the possible biological damage correlated with the physical features of the responsible HZE-particle. In the different systems the degree of damage depended on whether the hit cell was replaceable or not. A high sensitivity to HZE-particle bombardment was observed on Artemia salina eggs; 90% of the embryos, which were induced to develop from hit eggs, died at different developmental stages. Malformations of the abdomen or the extremities of the nauplius were frequently induced. In contrast, the growth of hit Vicia faba radiculae and the germination of hit Arabidopsis thaliana seeds and hit Bacillus subtilis spores were not influenced remarkably. But there was an increase in multicaulous plants and a reduction in the outgrowth of the bacterial spores. In addition, information was obtained on the fluence of the HZE-particles, on their spectrum of charge and energy loss, and on the absorption by the Apollo spacecraft and the Biostack material itself. This will help to improve knowledge concerning radiation conditions inside of spacecrafts, necessary to secure a maximum possible protection to the astronauts. PMID- 11887917 TI - Skylab experiment M-092: results of the first manned mission. AB - Blood pressure at 30-sec intervals, heart rate, and percentage increase in leg volume continuously were recorded during a 25-min protocol in the M092 Inflight Lower Body Negative Pressure (LBNP) experiment carried out in the first manned Skylab mission. These data were collected during six tests on each crewman over a 5-month preflight period. The protocol consisted of a 5-min resting control period, 1 min at -8, 1 min at -16, 3 min at -30, 5 min at -40, and 5 min at -50 mm Hg LBNP. A 5-min recovery period followed. Inflight tests were performed at approximately 3-day intervals through the 28-day mission. Individual variations in cardiovascular responses to LBNP during the preflight period continued to be demonstrated in the inflight tests. Measurements of the calf indicated that a large volume of fluid was shifted out of the legs early in the flight and that a slower decrease in leg volume, presumably due to loss of muscle tissue, continued throughout the flight. Resting heart rates tended to be low early in the flight and to increase slightly as the flight progressed. Resting blood pressure varied but usually was characterized by slightly elevated systolic blood pressure, lower diastolic pressure, and higher pulse pressures than during preflight examinations. During LBNP inflight a much greater increase in leg volume occurred than in preflight tests. Large increases occurred even at the smallest levels of negative pressure, suggesting that the veins of the legs were relatively empty at the beginning of the LBNP. The greater volume of blood pooled in the legs was associated with greater increases of heart rate and diastolic pressure and larger falls of systolic and pulse pressure than seen in preflight tests. The LBNP protocol represented a greater stress inflight, and on three occasions it was necessary to stop the test early because of impending syncopal reactions. LBNP responses inflight appeared to predict the degree of postflight orthostatic intolerance. Postflight responses to LBNP during the first 48 hours were characterized by marked elevations of heart rate and instability of blood pressure. In addition, systolic and diastolic pressures were typically elevated considerably both at rest and also during stress. The time required for cardiovascular responses to return to preflight levels was much slower than in the case of Apollo crewmen. PMID- 11887918 TI - Mineral and nitrogen balance study: results of metabolic observations on Skylab II 28-day orbital mission. AB - Prediction that the various stresses of flight, particularly weightlessness, would bring about significant derangements in the metabolism of the musculoskeletal system has been based on various observations of long-term immobilized or inactive bed rest. The only attempt at controlled measurement of metabolic changes in space prior to Skylab, a study during the 14-day Gemini VII flight, revealed rather modest losses of important elements. The three astronauts of Skylab II consumed a planned day-by-day, quite constant, dietary intake of major metabolic elements in mixed foods and beverages and provided virtually complete collections of excreta for 31 days preflight, during the 28 days inflight, and for 17 days postflight. Analyses showed that, in varying degree among the crewmen, urinary calcium increased gradually during flight in a pattern similar to that observed in bed-rest studies: the mean plateau peak of urinary calcium excretion in the latter part of flight was double preflight levels. Fecal calcium excretion did not change significantly, but calcium balance, owing to the urinary calcium rise, became either negative or less positive than in preflight measurement. Increased excretion and negative balance of nitrogen and phosphorus indicated appreciable loss of muscle tissue in all three crewmen. Significant losses also occurred inflight in potassium, sodium, and magnesium. Based on the similarity in pattern and degree between these observations and those in bed rest of the losses in calcium, phosphorus, and nitrogen, musculoskeletal integrity would not be threatened in space flights of up to at least 3 months. However, if similar changes occur, indicative of continuing losses of these elements, in the planned Skylab flights for considerably more than 28 days, concern for capable musculoskeletal function should be serious for flights of very many months' duration, and greater research attention will need to be given to development of protective counter-measures. PMID- 11887919 TI - Postmission plasma volume and red-cell mass changes in the crews of the first two Skylab missions. AB - Red-cell mass determinations were performed before and after the first two Skylab missions. The data showed a 14% mean decrease in red-cell mass after the 28-day mission and a 12% mean decrease after the 59-day mission. The red-cell mass returned to premission levels more slowly after the shorter (28-day) than after the longer mission. Plasma volume decreases were found after each mission. with the crew from the longer mission showing the greater change (13% vs. 8.4%). Postmission decreases in red-cell mass and plasma volume have been a general finding in crewmen who return from short or long spaceflight. PMID- 11887920 TI - The Skylab sleep monitoring experiment: methodology and initial results. AB - The sleep monitoring experiment permitted an objective evaluation of sleep characteristics during the first two manned Skylab flights. Hardware located onboard the spacecraft accomplished data acquisition, analysis, and preservation, thereby permitting near-real-time evaluation of sleep during the flights and more detailed postmission analysis. The crewman studied during the 28-Day Mission showed some decrease in total sleep time an increase in the percentage of Stage 4 sleep, while the subject in the 59-Day Mission exhibited little change in total sleep time and a small decrease in Stage 4 and REM sleep. Some disruption of sleep characteristics was seen in the final days of both missions, and both subjects exhibited decreases in REM-onset latency in the immediate postflight period. The relatively minor changes seen were not of the type nor magnitude which might be expected to be associated with significant degradation of performance capability. PMID- 11887921 TI - Skylab task and work performance (experiment M-151--time and motion study). AB - The primary objective of Experiment M151 was to study by means of time and motion analytic techniques the inflight adaptation of Skylab crewmen to a variety of task situations involving different types of activity. A parallel objective was to examine astronaut inflight performance for any behavioral stress effects associated with the working and living conditions of the Skylab environment. Training data provided the basis for comparison of preflight and inflight performance. Efficiency was evaluated through the adaptation function, namely, the relation of performance time over task trials. The results indicate that the initial changeover from preflight to inflight (or, from 1-G to zero-G) was accompanied by a substantial increase in performance time for most work and task activities. Equally important was the finding that crewmen adjusted rapidly to the weightless environment and became proficient in developing techniques with which to optimize task performance. By the end of the second inflight trial, most of the activities were performed almost as efficiently as on the last preflight trial. In addition, the analysis demonstrated the sensitivity of the adaptation function to differences in task and hardware configuration. The function was found to be more regular and less variable inflight than preflight. Translation and control of masses (large or small) were accomplished easily and efficiently through the rapid development of the arms and legs (and the entire body) as subtle guidance and restraint systems. Finally, the adaptation function provided no evidence of behavioral stress effects attributable to the Skylab environment. PMID- 11887922 TI - Skylab experiment M-171 "Metabolic Activity"--results of the first manned mission. AB - The experiment was performed to ascertain whether man's ability to perform mechanical work would be altered as a result of exposure to the weightless environment. Skylab II crewmen were exercised on a bicycle ergometer at loads approximating 25%, 50%, and 75% of their maximum oxygen uptake while their physiological responses were monitored. The results of these tests indicate that the crewmen had no significant decrement in their response to exercise during their exposure to zero gravity. Immediately postflight, however, all crewmen demonstrated an inability to perform the programmed exercise with the same metabolic effectiveness as they did both preflight and inflight. The most significant changes were elevated heart rates for the same work load and oxygen consumption (decreased oxygen pulse), decreased stroke volume, and decreased cardiac output at the same oxygen consumption level. It is apparent that the changes occurred inflight, but did not manifest themselves until the crewmen attempted to readapt to the 1-G environment. PMID- 11887923 TI - Wisconsin's BadgerCare program offers innovative approach for family coverage. AB - Wisconsin's BadgerCare program is viewed by many as a model for how other states could pursue comprehensive health insurance coverage for lower income families. The program provides health insurance to working families - both children and their parents - and seeks to eliminate barriers to successful employment by providing a transition for families from welfare to private insurance. BadgerCare's success is founded on its family coverage approach, its single point of entry and administrative seamlessness, and the political commitment to the program from Governor Tommy G. Thompson. In 1999 and 2000, New York, New Jersey, and the Clinton administration recognized the importance of a family-based approach to children's coverage by proposing, and in the states' case, implementing some variation of the Wisconsin model. Other states have indicated interest in covering families and look to the flexibility of the Health Care Financing Administration's (HCFA) recent ruling on 1115 demonstration projects to cover parents using the enhanced State Children's Health Insurance Program (SCHIP) match. This case study details the BadgerCare program and its impact on the uninsured in Wisconsin, including how the program approaches enrolling families, how family coverage is financed, how the program partners with private insurance, and what cost-sharing obligations exist. PMID- 11887924 TI - Section 1115 waivers and budget neutrality: using Medicaid funds to expand coverage. PMID- 11887925 TI - Full-cost buy-ins: an overview of state experience. PMID- 11887926 TI - Medicaid disease management: seeking to reduce spending by promoting health. PMID- 11887927 TI - Welfare of the individual and the group: Malthus and externalities. PMID- 11887928 TI - Darwin fallen among political economists. PMID- 11887929 TI - World food trends: a neo-Malthusian prospect? PMID- 11887931 TI - Potentiation and prolongation of the insulinotropic action of glucagon-like peptide 1 by methyl pyruvate or dimethyl ester of L-glutamic acid in a type 2 diabetes animal model. AB - Methyl pyruvate and the dimethyl ester of L-glutamic acid were administered intravenously, as a primed constant infusion (1.0-2.0 micromol followed by 0.5 1.0 micromol/min, both expressed per gram of body wt), in adult rats that had been injected with streptozotocin during the neonatal period. Each ester augmented plasma insulin concentration and potentiated and/or prolonged the insulinotropic action of glucagon-like peptide 1 (GLP-1) injected intravenously (5 pmol/g of body wt) at min 5 of the test. It is proposed, therefore, that suitable nonglucidic nutrients, susceptible to bypassing the site-specific defects of D-glucose transport and metabolism responsible for the preferential impairment of the B-cell secretory response to D-glucose in non-insulin-dependent diabetes, could be used to optimize the insulinotropic action of GLP-1. PMID- 11887930 TI - Calmodulin and a cyclic nucleotide-dependent protein kinase facilitate the prolactin-induced increase in tyrosine hydroxylase activity in tuberoinfundibular dopaminergic neurons. AB - Many aspects of tuberoinfundibular dopaminergic neuronal function are increased by elevated prolactin (PRL) levels, including the activity of tyrosine hydroxylase, the rate-limiting enzyme in the biosynthesis of dopamine. This study evaluated the roles of calmodulin, cyclic nucleotide-dependent protein kinase, and calcium/calmodulin-dependent protein kinase II in the PRL-induced increase in tyrosine hydroxylase activity. Ovariectomized rats were treated with haloperidol or ovine PRL (oPRL) for 20-30 h before the experiment, respectively. Treatment with haloperidol increased circulating PRL levels 8-fold and tyrosine hydroxylase activity in the stalk-median eminence 1.8-fold. Treatment with oPRL increased tyrosine hydroxylase activity 1.9-fold. W-7, a calmodulin antagonist, reversed both the haloperidol- and oPRL-induced increase in tyrosine hydroxylase activity to control levels. H-8, a cyclic nucleotide-dependent protein kinase inhibitor, also reversed the haloperidol induced increase in tyrosine hydroxylase activity. KN62, a selective calcium/calmodulin-dependent protein kinase II inhibitor, attenuated the haloperidol-induced increase in tyrosine hydroxylase activity, but KNO4, a structurally related control compound, had no effect. By contrast, the oPRL- and haloperidol-induced increases in tyrosine hydroxylase activity were not altered by KN93, a selective calcium/calmodulin-dependent protein kinase II inhibitor. These data indicate that calmodulin and a cyclic nucleotide-dependent protein kinase contribute to the PRL-induced increase in tyrosine hydroxylase activity, but the role of calcium/calmodulin-dependent protein kinase II is still unclear. PMID- 11887932 TI - Urinary vasodilator and vasoconstrictor angiotensins during menstrual cycle, pregnancy, and lactation. AB - Since normal human pregnancy is characterized by normotension in the face of an increased renin-angiotensin-aldosterone system (RAAS), we evaluated the temporal pattern of urinary excretion of a novel vasodilator within this system, angiotensin-(1-7) (Ang-[1-7]), during the menstrual cycle, pregnancy, and lactation. The urinary profiles of Ang I, Ang II, human chorionic gonadotropin, 17beta-estradiol, and progesterone were also determined. During the menstrual cycle, urinary Ang-(1-7) and Ang II remained stable (mean cycle value: 94.6 +/- 11.3 and 11.4 +/- 1.1 pmol/g of creatinine, respectively) in nine females. In 10 normal pregnant women, urinary Ang-(1-7) and Ang II increased throughout gestation, averaging 1499.8 +/- 310 and 224.4 +/- 58 pmol/g of creatinine, respectively (p < 0.05) at wk 35 and falling during lactation to 394.0 +/- 95 and 65.7 +/- 20 pmol/ g of creatinine (p < 0.05), respectively. The Ang-(1-7)/Ang II ratio was unchanged in the different reproductive periods. During the menstrual cycle, Ang II and Ang-(1-7) correlated with 17beta-estradiol and progesterone using multivariate analysis (r = 0.31, p < 0.001) and r = 0.28, p < 0.02, respectively). During gestation, 17beta-estradiol and progesterone correlated with urinary Ang-(1-7) (r = 0.48, p < 0.001 and r = 0.47, p < 0.001, respectively) and Ang II (r = 0.24, p < 0.03 and r = 0.25, p < 0.03, respectively); by multiple regression, only Ang-(1-7) correlated with both steroids (r = 0.49,p < 0.001). The progressive rise of Ang-(1-7) throughout gestation, probably modulated by estrogen and progesterone, suggests a physiologic counterregulation within the RAAS. PMID- 11887934 TI - Autocrine regulation of prolactin secretion by endothelins: a permissive role for estradiol. AB - We have previously found that lactotrophs express and secrete endothelin-like peptides that influence prolactin (PRL) secretion in an autocrine fashion. We have also observed that the incidence of endothelin-immunoreactive lactotrophs is markedly affected by ovarian steroids. In this study, we examined how the ovarian steroid background determines the efficiency of the endothelin-mediated autocrine feedback regulation of PRL secretion. Ovariectomized adult female rats were used throughout these studies. Steroid replacements were made by sc implantation of Silastic capsules immediately following ovariectomy. Eight to 10 wk later, three animals from each treatment group (no steroid control, estradiol, progesterone, estradiol plus progesterone) were sacrificed by decapitation, and the anterior pituitary cells were enzymatically dispersed using collagenase and hyaluronidase. A PRL-specific reverse hemolytic plaque assay was used to measure PRL secretion at the single-cell level. BQ123, a synthetic cyclic pentapeptide with distinctive endothelin-A receptor antagonist quality, caused only a modest elevation of PRL secretion in the control group. Endothelin antagonism did not affect PRL secretion in cells obtained from progesterone-implanted animals. Endothelin antagonism did, however, increase overall PRL secretion in the estradiol and estradiol plus progesterone groups by five- and threefold, respectively. Frequency distribution of PRL plaques in these same two BQ123-treated groups revealed two subpopulations, indicating that lactotrophs differ in their response to endogenous endothelin feedback and that this difference is steroid dependent. These observations clearly suggest that the ovarian steroid milieu (estrogens in particular) can have a profound influence on the self-regulatory mechanisms of lactotrophs. Our results also emphasize that endogenous endothelins may play an important role in the negative feedback regulation of PRL secretion in female rats. PMID- 11887935 TI - Growth hormone therapy during neonatal hypoxia in rats: body composition, bone mineral density, and insulin-like growth factor-1 expression. AB - Hypoxia from birth results in a decrease in body weight gain, body size, and bone mineral density (BMD). The purpose of the present study was to determine whether short-term administration of growth hormone (GH) (rat GH; 100 microg/d) could attenuate some of these effects of neonatal hypoxia. Rat pups (with their lactating dams) were exposed to hypoxia (vs normoxic control) from birth. Hypoxia was continued until 14 d of age, with rat GH (vs vehicle control) administered daily. Hypoxia significantly inhibited body weight gain; GH therapy did not reverse this effect. GH therapy did reverse the inhibitory effect of hypoxia on tail length but not on body length. Hypoxia decreased BMD analyzed by dual X-ray absorptiometry (DXA); this effect was not reversed by GH therapy. Both GH therapy and hypoxia decreased the percentage of body fat analyzed by DXA, the effects of which were additive when combined. There were minimal effects of hypoxia and GH therapy on plasma insulin-like growth factor-1 (IGF-1), IGF-binding protein-3, and hepatic IGF-1 mRNA expression. We conclude that some of the effects of hypoxia on body habitus are reversed by GH therapy, but that short-term GH therapy did not prevent a loss of BMD. GH therapy for more than 14 days may be necessary to appreciate fully its potential in the treatment of the sequelae of neonatal hypoxia. PMID- 11887933 TI - Expression of cyclooxygenase enzymes in rat hypothalamo-pituitary-adrenal axis: effects of endotoxin and glucocorticoids. AB - Prostaglandins play a key role in mediating the hypothalamo-pituitary adrenocortical (HPA) responses to immune insults. This study aimed to provide some insight into the relative contributions of the constitutive and inducible forms of cyclooxygenase (COX-1 and COX-2) to the generation of these prostanoids by examining the effects of (1) endotoxin treatment on the expression of COX-1 and COX-2 mRNAs in the various components of the HPA axis in control and glucocorticoid pretreated rats, and (2) selective inhibition of COX-2 on the production of corticosterone by adrenal tissue in vitro. Endotoxin caused a marked rise in COX-2 mRNA in the adrenal gland that was evident 3 and 6 h after the injection and was prevented by pretreatment with dexamethasone. It also induced a modest increase in COX-2 mRNA in the hypothalamus but not in the hippocampus or anterior pituitary gland. By contrast, COX-1 mRNA was largely unaffected by the drug treatments in all tissues studied. In vitro the selective COX-2 inhibitor SC-236 caused a marked reduction in adrenocorticotropic hormone driven corticosterone release, as did the nonselective COX inhibitor, indomethacin. These results support a role of COX-2 in the manifestation of the HPA responses to endotoxin, particularly within the adrenal gland. PMID- 11887936 TI - Glycogen storage disease lb and Crohn colitis in a young woman. AB - The occurrence of inflammatory bowel disease in patients with glycogen storage disease lb is rare (GSD-lb). We present the case of a young woman with the diagnosis of GSD-lb Crohn-like colitis developed at age 22. Clinical evaluation revealed severe malnutrition, secondary amenorrhea, leukopenia, neutropenia, dysfunctions of phagocytosis, and subtotal stenosis of the ascending colon. Right hemicolectomy was performed and pathohistologic analysis of the resected bowel showed chronic bowel inflammation consistent with Crohn disease. Clinical status of the patient markedly improved after surgery. PMID- 11887937 TI - Epidermal growth factor receptor and tyrosine phosphorylation of estrogen receptor. AB - Activation of estrogen receptor-alpha (ERalpha) by growth factors in the absence of estrogen is a well-documented phenomenon. To study further this process of ligand-independent receptor activation, COS-7 cells without ER were transfected with both ER and epidermal growth factor receptor (EGFR). In the absence of estrogen, epidermal growth factor (EGF) stimulated rapid tyrosine phosphorylation of ER in transfected COS-7 cells. Similarly, in MCF-7 breast cancer cells that have natural expression of ER and EGFR, EGF promoted acute phosphorylation of serine and tyrosine residues in ER, and a direct interaction between ER and EGFR after treatment with EGF was found. In confirmation of a direct interaction between ER and EGFR, activation of affinity-purified EGFR tyrosine kinase in vitro stimulated the phosphorylation of recombinant ER. The cross-communication between EGFR and ER appears to promote significant stimulation of cell proliferation and a reduction in the apoptotic loss of those cells that express both receptor signaling pathways. However, COS-7 cells transfected with both ER and EGFR show minimal stimulation of classical estrogen response element (ERE) dependent transcriptional activity after stimulation by EGF ligand. This suggests that the proliferative and antiapoptotic activity of EGF-induced ER activation may be dissociated from ERE-dependent transcriptional activity of the ER. PMID- 11887938 TI - Prostaglandin F2alpha-activated protein kinase Calpha phosphorylates myristoylated alanine-rich C kinase substrate protein in bovine luteal cells. AB - Prostaglandin F2alpha (PGF2alpha)-induced secretion of oxytocin by the bovine corpus luteum involves the phosphorylation of a unique protein kinase C (PKC) substrate, myristoylated alanine-rich C kinase substrate (MARCKS) protein. This study was conducted to determine the specific PKC isoform engaged in phosphorylation of MARCKS protein in bovine luteal cells. In experiment 1, dispersed luteal cells recovered from the corpus luteum on d 8 of the estrous cycle were preincubated with [32P] orthophosphate and then exposed to PGF2alpha alone or in combination with PKC inhibitors. Autoradiography and densitometry of Western blots revealed that MARCKS protein was phosphorylated by a conventional PKC (cPKC) isoform. Experiment 2 was conducted to identify the specific cPKC isoform that phosphorylates MARCKS protein in luteal cells. Corpora lutea were removed from control and PGF2alpha-treated heifers on d 8 of the cycle, and PKC isoforms associated with membrane and cytosolic fractions were determined. Treatment with PGF2alpha increased membrane concentrations of PKCalpha within 5 min after treatment (p < 0.005). Collectively, these data suggest that phosphorylation of MARCKS protein coinciding with oxytocin secretion is mediated by PKCalpha. PMID- 11887939 TI - Expression of Hsp70-2 in unilateral cryptorchid testis of rhesus monkey during germ cell apoptosis. AB - We investigated the possible role of Hsp70-2 in germ cell apoptosis induced by heat stress in monkey unilateral cryptorchid testis. The study focused on in situ analysis of the testicular cell DNA fragmentation and on the possible relationship between Hsp70-2 expression and germ cell apoptosis. The TUNEL result showed that most of the germ cells were labeled in the cryptorchid testis on d 5 after induction of cryptorchidism; that with most of the apoptotic germ cells depleted, only a few germ cells were labeled on d 10; and that almost no apoptotic signal was observed in the cryptorchid testis on d 15 and thereafter. This indicates that the increasing germ cell degeneration in cryptorchid testis may take the form of apoptosis. Using in situ hybridization, immunohistochemistry, and Northern blot, we examined the changes of Hsp70-2 expression in the monkey cryptorchid testis. The level of Hsp70-2 mRNA decreased slightly, while the expression of HSP70-2 protein was almost unchanged at the early stage of germ cell apoptosis in the cryptorchid testis on d 5 and dropped dramatically along with the loss of apoptotic germ cells in the cryptorchid testis on d 10 after operation. It is therefore suggested that Hsp70-2 might not take part in inhibiting the apoptosis of germ cells at the early stage during operation-induced cryptorchid testis, and that Hsp70-2 gene does not belong to the immediate early related gene responsible for germ cell apoptosis induced by heat stress. PMID- 11887941 TI - Mammographic density: a breast cancer risk factor or diagnostic indicator? PMID- 11887940 TI - Total parenteral nutrition modulates hormone release by stimulating expression and activity of inducible nitric oxide synthase in rat pancreatic islets. AB - The expression and activities of constitutive nitric oxide synthase (cNOS) and inducible nitric oxide synthase (iNOS) in relation to insulin and glucagon secretory mechanisms were investigated in islets isolated from rats subjected to total parenteral nutrition (TPN) for 10 d. TPN is known to result in significantly increased levels of plasma lipids during the infusion time. In comparison with islets from freely fed control rats, islets taken from TPN rats at d 10 displayed a marked decrease in glucose-stimulated insulin release (4.65 +/- 0.45 ng/[islet x h] vs 10.25 +/- 0.65 for controls) (p < 0.001) accompanied by a strong iNOS activity (18.3 +/- 1.1 pmol of NO/[min x mg of protein]) and a modestly reduced cNOS activity (11.3 +/- 3.2 pmol of NO/[min x mg of protein] vs 17.7 +/- 1.7 for controls) (p < 0.01). Similarly, Western blots showed the expression of iNOS protein as well as a significant reduction in cNOS protein in islets from TPN-treated rats. The enhanced NO production, which is known to inhibit glucose-stimulated insulin release, was manifested as a strong increase in the cyclic guanosine 5'-monophosphate content in the islets of TPN-treated rats (1586 +/- 40 amol/islet vs 695 +/- 64 [p < 0.001] for controls). Moreover, the content of cyclic adenosine monophosphate (cAMP) was greatly increased in the TPN islets (80.4 +/- 2.1 fmol/islet vs 42.6 +/- 2.6 [p < 0.001] for controls). The decrease in glucose-stimulated insulin release was associated with an increase in the activity of the secretory pathway regulated by the cAMP system in the islets of TPN-treated rats, since the release of insulin stimulated by the phosphodiesterase inhibitor isobutylmethylxanthine was greatly increased both in vivo after iv injection and after in vitro incubation of isolated islets. By contrast, the release of glucagon was clearly reduced in islets taken from TPN treated rats (33.5 +/- 1.5 pg/[islet x h] vs 45.5 +/- 2.2 for controls) (p < 0.01) when islets were incubated at low glucose (1.0 mmol/L). The data show that long-term TPN treatment in rats brings about impairment of glucose-stimulated insulin release, that might be explained by iNOS expression and a marked iNOS derived NO production in the beta-cells. The release of glucagon, on the other hand, is probably decreased by a direct "nutrient effect" of the enhanced plasma lipids. The results also suggest that the islets of TPN-treated rats have developed compensatory insulin secretory mechanisms by increasing the activity of their beta-cell cAMP system. PMID- 11887943 TI - Vasospasm model of a rabbit common carotid artery for endovascular research. AB - RATIONALE AND OBJECTIVES: Endovascular treatments such as balloon angioplasty and injection of vasodilators play an important role in cerebral vasospasm refractory to first-line therapies. Experimental studies with intracranial arteries of animals have some limitations, however, because in most cases endovascular devices are too large and inflexible to catheterize these arteries. The purpose of this study was to document the natural course of a vasospasm model using a rabbit common carotid artery. MATERIALS AND METHODS: The common carotid arteries of 24 New Zealand white rabbits were used. Autologous blood was placed around the isolated artery and kept in place with a silicone cuff. The time course of vasoconstriction after blood clot placement was examined with angiography and with histopathologic evaluation. RESULTS: Angiographic vasoconstriction was prominent on day 2, less obvious on day 9, and absent on day 30. At histopathologic evaluation, folding of the endothelial surface, corrugation of the internal elastic lamina, and thickening of the arterial wall were observed in the group with blood clot placements around the artery on day 2 and were nearly absent on day 30. CONCLUSION: This model may be used for the study of endovascular treatments of vasospasm. PMID- 11887942 TI - Multimodality diagnosis of liver tumors: feature analysis with CT, liver-specific and contrast-enhanced MR, and a computer model. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to measure and to clarify the diagnostic contributions of image-based features in differentiating benign from malignant and hepatocyte-containing from non-hepatocyte-containing liver lesions. MATERIALS AND METHODS: Six experienced abdominal radiologists each read images from 146 cases (including a contrast material-enhanced computed tomographic [CT] scan and contrast-enhanced and unenhanced magnetic resonance [MR] images) following a checklist-questionnaire requiring them to rate quantitatively each of as many as 131 image features and then reported on each of the two differentiations. The diagnostic value of each feature was assessed, and linear discriminant analysis was used to develop statistical prediction rules (SPRs) for merging feature data into computerized "second opinions." For the two differentiations, accuracy (area under the receiver operating characteristic curve [Az]) was then determined for the radiologists' readings by themselves and for each of three SPRs. RESULTS: Thirty-seven candidate features had diagnostic value for each of the two differentiations (a slightly different feature set for each). Radiologists' performance at both differentiations was excellent (Az = 0.929 [benign vs malignant] and 0.926 [hepatocyte-containing vs non-hepatocyte containing]). Performance of the SPR that operated on the features from all modalities together was better than that of radiologists (Az = 0.936 [benign vs malignant] and 0.951 [hepatocyte-containing vs non-hepatocyte-containing]), but this difference was of marginal statistical significance (P = .11). Contrast enhanced MR imaging and contrast-enhanced CT each made significant adjunctive contributions to accuracy compared with unenhanced MR imaging alone. CONCLUSION: Many CT- and MR imaging-based features have diagnostic value in differentiating benign from malignant and hepatocyte-containing from non-hepatocyte-containing liver lesions. Radiologists could also benefit from the fully informed SPR's "second opinions." PMID- 11887944 TI - Changes in the fibrinolytic system during angiography with ionic and with nonionic contrast media. AB - RATIONALE AND OBJECTIVES: The purpose of the study was to evaluate and compare changes in some parameters of the fibrinolytic system caused by the use of ionic and nonionic contrast media during angiography in certain groups of patients. MATERIALS AND METHODS: Angiographic diagnostic procedures were performed in 126 patients (male and female) clinically suspected of having kidney cancer (38 patients), arteriosclerotic occlusive disease of lower extremities (44 patients), or dissection of cerebral artery (44 patients). The control group included 12 patients with clinical symptoms of the disease in whom angiographic examination excluded the presence of cerebral artery dissection or kidney cancer. Patients were randomly assigned to receive either an ionic (diatrizoate sodium) or a nonionic (iopromide) contrast medium. Immediately before and 30 minutes after administration, venous blood samples were obtained to determine select parameters of the hemostatic system. RESULTS: There were no significant differences in the fibrinolytic parameters within the control group after contrast medium administration. The nonionic contrast medium (iopromide) caused a decrease in fibrinolytic activity in the patients, unlike the controls, which was particularly pronounced among the patients undergoing renal angiography. CONCLUSION: The use of contrast media in some groups of patients led to transient changes in the fibrinolytic system. These results indicate that ionic contrast media should be used during angiographic procedures in patients at increased risk for thrombotic complications. PMID- 11887945 TI - Estimation in medical imaging without a gold standard. AB - RATIONALE AND OBJECTIVES: In medical imaging, physicians often estimate a parameter of interest (eg, cardiac ejection fraction) for a patient to assist in establishing a diagnosis. Many different estimation methods may exist, but rarely can one be considered a gold standard. Therefore, evaluation and comparison of different estimation methods are difficult. The purpose of this study was to examine a method of evaluating different estimation methods without use of a gold standard. MATERIALS AND METHODS: This method is equivalent to fitting regression lines without the x axis. To use this method, multiple estimates of the clinical parameter of interest for each patient of a given population were needed. The authors assumed the statistical distribution for the true values of the clinical parameter of interest was a member of a given family of parameterized distributions. Furthermore, they assumed a statistical model relating the clinical parameter to the estimates of its value. Using these assumptions and observed data, they estimated the model parameters and the parameters characterizing the distribution of the clinical parameter. RESULTS: The authors applied the method to simulated cardiac ejection fraction data with varying numbers of patients, numbers of modalities, and levels of noise. They also tested the method on both linear and nonlinear models and characterized the performance of this method compared to that of conventional regression analysis by using x axis information. Results indicate that the method follows trends similar to that of conventional regression analysis as patients and noise vary, although conventional regression analysis outperforms the method presented because it uses the gold standard which the authors assume is unavailable. CONCLUSION: The method accurately estimates model parameters. These estimates can be used to rank the systems for a given estimation task. PMID- 11887946 TI - Mammographic tissue, breast cancer risk, serial image analysis, and digital mammography. Part 1. Tissue and related risk factors. AB - This work is presented as a sequence of two parts. In this leading section, a review of the breast tissue-risk research is provided. Although controversy remains, there is substantial evidence indicating that dense mammographic tissue (a) is a breast cancer risk factor that is at least similar, if not greater, in magnitude with the other known breast cancer risk factors and (b) may be a partial biomarker for some of the other risk factors. Understanding these influences may provide a mechanism for measuring the dynamics of breast cancer risk. The totality of this work is to provide support for an automated serial mammography study under way at the authors' institution, where digital mammographic images are acquired with a full-field digital mammography system. This is a filmless imaging system, where the image is acquired in digital format. This electronic imaging acquisition system provides a prime opportunity to easily couple and manipulate the image data with patient information such as risk probability analysis or other pertinent personal history data for improved automated decision making. In this leading section, the main focus is on understanding elements that will assist in fusing risk probability analysis with automated computer-aided diagnosis. The evidence indicates that there are many factors that influence breast tissue at any given time and thus have the ability to alter the associated radiographic image appearance over time. At the initiation of the serial study it was clear that the authors did not fully understand the nature of the problem: automatically comparing similar mammographic scenes acquired at different times. In the second part of this sequence, the more time-related tissue influences are reviewed. PMID- 11887947 TI - Mammographic tissue, breast cancer risk, serial image analysis, and digital mammography. Part 2. Serial breast tissue change and related temporal influences. AB - The work presented herein is the second part of a review of breast tissue-related cancer-risk research. Briefly, in part 1, the tissue-risk research is discussed. In this part, factors that influence temporal breast tissue change are reviewed. Since breast composition is correlated with some of the known risk factors, understanding these influences may provide a mechanism for measuring the dynamics of breast cancer risk. The purpose of this work is to provide support for an automated serial mammography study under way at the authors' institution, where the digital mammographic images are acquired with a full-field digital mammography imaging system. At the initiation of the serial study, it was clear that the authors did not fully understand the nature of the problem: automatically comparing similar mammographic scenes acquired at different times. The evidence indicates that there are many factors that influence breast tissue at any given time and thus have the ability to alter the associated radiographic appearance over time. In general, the topics considered herein include aging; involution; breast development; exogenous and endogenous hormonal interactions such as hormone replacement therapy, oral contraceptive use, and menstrual timing; screening sensitivity issues and interval cancers; tumor growth rates; sojourn times; and lifestyle factors such as diet and exercise. Throughout this work, commentaries and suggestive strategies for automated serial image analysis are provided. PMID- 11887948 TI - Low-grade siderotic dysplastic nodules: determination of premalignant lesions on the basis of vasculature phenotype. AB - RATIONALE AND OBJECTIVES: The authors performed this study to determine whether, on the basis of the vascular profile, low-grade siderotic dysplastic nodules are premalignant lesions. MATERIALS AND METHODS: The authors used a monoclonal antibody specific for smooth muscle actin to stain 18 siderotic low-grade dysplastic nodules (mean size, 0.7 cm) from nine patients. Two pathologists counted the number of unpaired arteries per high-power field in siderotic dysplastic nodules and background siderotic regenerative nodules by using two techniques (conventional and hot spot). RESULTS: The number of unpaired arteries seen with the conventional counting technique in low-grade siderotic dysplastic nodules (range, 1-14; mean, 3.8) was significantly greater (P = .004) than that seen in background siderotic regenerative nodules (range, 0-3; mean, 1.2). Similarly, the number of unpaired arteries seen with the hot spot technique in low-grade siderotic dysplastic nodules (range, 0-14; mean, 5.2) was significantly greater (P = .005) than that seen in background siderotic regenerative nodules (range, 0-6; mean, 1.9). CONCLUSION: On the basis of the vascular profile, low grade siderotic dysplastic nodules should be considered premalignant lesions. Further research is needed to help differentiate these lesions from siderotic regenerative nodules with magnetic resonance imaging. PMID- 11887949 TI - The integrated clinical year: a unique approach to training radiology residents at the University of New Mexico. PMID- 11887950 TI - Software-annotated, digitally photographed, and printed MR images: suitability for publication. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate whether digitally photographed, computer-annotated MR images produced by clinical radiologists and printed with an inexpensive photo printer are suitable for publication. MATERIALS AND METHODS: Laser prints of 20 magnetic resonance images of the brain were photographed with a 3-megapixel digital camera and annotated with arrows, arrowheads, and asterisks by using graphics software that incorporates vector support. Then, 5 x 7-inch glossy prints with white borders were made by using an inexpensive photo printer. These prints were compared with those produced of the same 20 images by members of the medical center's graphics department with professional scanning and printing equipment and annotated with conventional rub-on symbols. Eight radiologists evaluated image and annotation quality and overall suitability for publication. RESULTS: In all three categories, the images produced by radiologists outscored those produced by the graphics department. CONCLUSION: Digitally photographed, software-annotated MR images printed with an inexpensive photo printer are suitable for publication. PMID- 11887951 TI - Congressional update: Report from the Biomedical Imaging Program of the National Cancer Institute. American College of Radiology Imaging Network: The digital mammographic imaging screening trial--an update. PMID- 11887952 TI - "Enron-ing" health insurance? PMID- 11887953 TI - Health policy in the land of Oz. AB - Since the mid-1960s, the health sector has depended on the political process to shape the organization, delivery, and financing of medical care. When the health sector was in its infancy, using public policy to give form and substance to America's medical care was appropriate. Now, after nearly 35 years of legislative activity at the federal and state levels, American health care has become expensive, fragmented and complex-and the political process gridlocked. Can the necessary health reform efforts be stripped from an ineffective, politicized public policy process, and be accomplished from the bottom up by disenfranchised purchasers and providers of health care and their communities acting on their own initiative? PMID- 11887954 TI - Predicting response to regulatory change in the small group health insurance market: the case of association health plans and HealthMarts. AB - Lack of health insurance continues to be a concern for many people, even among those who are employed, and employees of small firms are much less likely to be insured than employees of larger firms. For several years, the U.S. Congress has considered legislation that would establish two new vehicles for offering health insurance coverage to small employers: association health plans (AHPs) and HealthMarts. In this paper, we present a model for estimating the impact the new entities would have on coverage and premiums in the small group health insurance market. The model produces a range of estimates based on assumptions, among others, about demand for insurance among small firms and their willingness to switch to less expensive, less generous benefit plans. We estimate that approximately 4.6 million people would obtain coverage through AHPs and HealthMarts, but fewer than half a million of them would be newly insured (based on 1999 population figures). Premiums would increase slightly for firms that continued to purchase coverage in the traditional market. PMID- 11887955 TI - Effects of "second generation" small group health insurance market reforms, 1993 to 1997. AB - In the mid-1990s, several state legislatures enacted a "second generation" of small group health insurance reforms that required guaranteed issue of all products and prohibited the use of health as a rating factor. We use data from two large employer surveys to compare the behavior of small business in nine states that adopted these reforms between 1993 and 1997 to the behavior of small business in 11 states and the District of Columbia, where neither of these small group health insurance market reforms existed prior to 1997 (N = 8,465 in 1993; N = 12,219 in 1997). Our analyses focus on several outcomes: health insurance offer and enrollment rates in any employer plan, and in an HMO plan; turnover in offer decisions; and premiums, variability in premiums, and the rate of change in premiums. Overall, we find no effect of small group reform on any of the outcomes; the sign of the effect is not consistent across reform states, the estimates rarely attain statistical significance, and they show no consistent pattern across the outcomes within each state. Therefore, predictions of the harm these regulations might cause to the market have not come to pass. On the other hand, proponents' hopes for a solution to low coverage rates among small businesses have not materialized either. PMID- 11887957 TI - Uncovering the missing Medicaid cases and assessing their bias for estimates of the uninsured. AB - General population surveys of health insurance coverage are thought to undercount Medicaid enrollment, which may bias estimates of the uninsured. This article describes the results of an experiment undertaken in conjunction with a general population survey in Minnesota. Responses to health insurance questions by a known sample of public program enrollees are analyzed to determine possible reasons for the undercount and the amount of bias introduced in estimates of uninsured people. While public program enrollees often misreport the type of coverage they have, the impact on estimates of those without insurance is negligible. Restrictions to generalizing the finding beyond this study are discussed. PMID- 11887956 TI - The impact of managed care on the use of outpatient mental health and substance abuse services in Puerto Rico. AB - This paper estimates the impact of managed care on use of mental health services by residents of low-income areas in Puerto Rico. A quasi-experimental design evaluates the impact of a low capitation rate on a minority population using three waves of data from a random community sample. Results indicate that two years after introducing managed care, privatization of mental health services had minimal impact on use. Advocates had hoped health care reform would increase access in comparison to access seen within the public system, while opponents feared profit motives would lead to decreased access. Neither forecast turned out to be correct. The question remains as to how to improve access for the poor with low capitation rates. PMID- 11887958 TI - Access and use by children on Medicaid: does state matter? AB - Although Medicaid is a central component of health care for children, the program is not uniform across the states. Using state and nationally representative data from the 1997 National Survey of America's Families (NSAF), a survey of the economic, health and social characteristics of children, nonaged adults and their families, we examine differences in access and use by children on Medicaid across 13 states, and compare those differences to national estimates. We find significant differences in access and use across the states for children on Medicaid. The characteristics of the children and their local health care environment explain some, but not all, of the state differences in access and use. PMID- 11887959 TI - Health-based risk adjustment: is inpatient and outpatient diagnostic information sufficient? AB - Adequate risk adjustment is critical to the success of market-oriented health care reforms in many countries. Currently used risk adjusters based on demographic and diagnostic cost groups (DCGs) do not reflect expected costs accurately. This study examines the simultaneous predictive accuracy of inpatient and outpatient morbidity measures and prior costs. DCGs, pharmacy cost groups (PCGs), and prior year's costs improve the predictive accuracy of the demographic model substantially. DCGs and PCGs seem complementary in their ability to predict future costs. However, this study shows that the combination of DCGs and PCGs still leaves room for cream skimming. PMID- 11887960 TI - Errors in data on hospital ownership. AB - In recent years, there has been increased public concern about changes in hospital ownership and consolidation in the hospital market. Numerous studies have examined the relationship between hospital ownership and prices, costs, organization, service mix, access to care and quality of care. However, the data on which these studies are based are often incomplete and inaccurate. In this paper, we examine several sources of data employed by researchers examining hospital ownership and discuss the degree of inaccuracy we found in each source. Substantial primary data collection is necessary to accurately account for changes in ownership and system affiliation. PMID- 11887961 TI - Modeling human epilepsies in mice. AB - Two categories of mouse models of human epilepsy are now contributing to the experimental analysis of inherited seizure disorders. The first type includes homologous genetic models arrived at in the classic way; the genes from human inherited epilepsy syndromes are cloned, and mice are recreated with functionally identical mutations. The second category involves the reverse strategy: mutating single genes in mice and determining whether the newly created nervous system develops epilepsy. These "gene-forward" models define specific candidate genes that can then be tested for possible involvement in human epilepsies. Spontaneous mutation of genes in mice and other species is also a source for candidate genes. As each of these genes and their physiologic functions is defined, the focus can shift to (a) fully characterizing the clinical epilepsy phenotype, (b) tracing the steps in the molecular pathogenesis of the disorder, and (c) pinpointing molecular targets for early intervention. Along with providing a unique opportunity to understand the mechanisms of inherited epileptogenesis, the mouse models serve as ideal biological test systems to search for novel therapeutic strategies. PMID- 11887962 TI - Genetics of the epilepsies. AB - Recent molecular insights into the human idiopathic epilepsies have suggested the central role of ligand-gated and voltage-gated ion channels in their etiology. So far, genes coding for sodium and potassium channel subunits as well as a nicotinic cholinergic receptor subunit have been identified for mendelian idiopathic epilepsies. In vitro and in vivo studies of mutations demonstrate functional changes, allowing new insights into mechanisms underlying hyperexcitability. Similarly, spontaneous murine epilepsy models have been associated with calcium channel molecular defects. The major challenge before us in understanding the genetics of the epilepsies is to identify genes for common forms of epilepsy following complex inheritance. Once such genes are discovered, the gene-gene-environmental interactions producing specific epilepsy syndromes can be explored. PMID- 11887963 TI - Overview of genetics for the clinician. PMID- 11887965 TI - Pharmacogenetics and antiepileptic drugs. AB - Responses among patients to antiepileptic drugs (AEDs) may be highly variable, with respect to both drug efficacy and safety. Pharmacogenetics addresses the genetic component of such patient variability. Differential response to phenytoin, for example, is related to interindividual genetic differences in the metabolic enzyme CYP2C9, and to a lesser extent, CYP2C19. However, no other AED responses have yet been linked conclusively to specific genes. Further understanding of the role of genes in AED response will depend on clinical investigations coupled with new information and technologies derived from the Human Genome Project. Once the DNA sequences involved in specific AED responses are understood, they can be used as the basis of clinical assays to predict the most likely response in each individual patient. The combination of clinical investigations, genomics, and emerging testing methodologies should lead to new tools for the effective management of epilepsy. PMID- 11887964 TI - Progress in the genetics of the partial epilepsies. AB - The importance of genetic contributions to the partial epilepsies is now well established. Evidence for this genetic contribution has come from familial aggregation studies, twin studies, positional cloning of specific genes that raise risk, and clinical descriptions of families. Familial aggregation studies are consistent in showing an increased risk of epilepsy in the relatives of patients with partial epilepsies that occur in the absence of environmental insults to the central nervous system. Susceptibility genes have been localized in five syndromes: autosomal dominant nocturnal frontal lobe epilepsy (20q, 1q, and 15q), autosomal dominant partial epilepsy with auditory features (10q), familial partial epilepsy with variable foci (22q), benign epilepsy of childhood with centrotemporal spikes (15q), and benign familial infantile convulsions (19q). In nocturnal frontal lobe epilepsy, the genes on chromosome 20q and 1q have been identified as subunits of the neuronal nicotinic acetylcholine receptor. PMID- 11887966 TI - Channelopathies: episodic disorders of the nervous system. PMID- 11887967 TI - Principles in pharmacogenetics. PMID- 11887968 TI - Potassium channels: how genetic studies of epileptic syndromes open paths to new therapeutic targets and drugs. AB - How can epilepsy gene hunting lead to better care for patients with epilepsy? Lessons may be learned from the progress made by identifying the mutated genes that cause Benign Familial Neonatal Convulsions (BFNC). In 1998, a decade of clinical and laboratory-based genetics work resulted in the cloning of the KCNQ2 potassium channel gene at the BFNC locus on chromosome 20. Subsequently, computer "mining" of public DNA databases allowed the rapid identification of three more brain KCNQ genes. Mutations in each of these additional genes were implicated as causes of human hereditary diseases: epilepsy (KCNQ3), deafness (KCNQ4), and, possibly, retinal degeneration (KCNQ5). Physiologists discovered that the KCNQ genes encoded subunits of the "M-channel," a type of potassium channel known to control repetitive neuronal discharges. Finally, pharmacologists discovered that retigabine, a novel anticonvulsant with a broad but distinctive efficacy profile in animal studies, was a potent KCNQ channel opener. These studies suggest that KCNQ channels may be an important new class of targets for anticonvulsant therapies. The efficacy of retigabine is currently being tested in multicenter clinical trials; identification of its molecular targets will allow it to be more efficiently exploited as a "lead compound." Cloned human KCNQ channels can now be expressed in cultured cells for "high-throughput" screening of drug candidates. Ongoing studies of the KCNQ channels in humans and animal models will refine our understanding of how M-channels control excitability at the cellular, network, and behavioral levels, and may reveal additional targets for therapeutic manipulation. PMID- 11887969 TI - Gene therapy for coronary heart disease. PMID- 11887970 TI - Clinical trends in atrial fibrillation at the turn of the millenium. PMID- 11887971 TI - Effects of intramyocardial injection of phVEGF-A165 as sole therapy in patients with refractory coronary artery disease--12-month follow-up: angiogenic gene therapy. AB - OBJECTIVE: To test the safety and bioactivity of phVEGF-A165 after intramyocardial injection during 12-month follow-up. DESIGN: Open-labelled study. SUBJECTS: Inclusion criteria were angina pectoris, Canadian Cardiovascular Society (CCS) class III-IV, unamenable to further revascularization, ejection fraction (EF) >30%, perfusion defects extending over >10% of the anterolateral left ventricle wall detectable with adenosine single photon emission computerized tomography (SPECT) and at least one patent vessel visible by coronary angiography. Seven of 39 patients referred for gene therapy were included. INTERVENTION: Via a mini-thoracotomy under general anaesthesia. phVEGF-A165 was injected directly into the myocardium at four sites in the anterolateral region of the left ventricle. RESULTS: Operative procedures were uneventful. Perioperative release of myocardial markers and electrocardiogram (ECG) changes were detected in two patients. There were no perioperative deaths but one patient died 7 months postoperatively because of myocardial infarction. Plasma vascular endothelial growth factor (VEGF)-A levels increased two to threefold peaking 6 days postoperatively (P < 0.004) and returning to baseline by day 30. A significant reduction in angina pectoris was reported. The CCS class improved from 3.3+/-0.2 to 1.9+/-0.3 (P < 0.01) and nitroglycerine intake decreased from 39+/-15 to 12+/-5 tablets week(-1) (P < 0.001) 2 months after gene transfer. Improvements remained after 12 months when nitroglycerine consumption approached zero. Improved myocardial function in the phVEGF-A165 injection region was documented in all patients (P < 0.016) by tissue velocity imaging (TVI). Reduced reversible ischaemia was detected by adenosine SPECT in four patients. Improved collateralization was detected in four patients with coronary angiography. CONCLUSION: Intramyocardial injection of phVEGF-A165 is safe and may lead to improved myocardial perfusion and function with longstanding symptomatic relief in end-stage angina pectoris. Based on these results this therapeutic potential is being tested in a double-blind placebo controlled multicentre trial. PMID- 11887972 TI - Hospitalizations for atrial fibrillation in the general male population: morbidity and risk factors. AB - OBJECTIVES: To analyse the incidence, prevalence, aetiology, risk factors and prognosis of hospitalizations for atrial fibrillation. SUBJECTS: A random population sample of 7495 men aged 47-55 years was first examined in 1970-73. During follow-up until 1996 (mean 25.2 years) 754 men were hospitalized with a diagnosis of atrial fibrillation. RESULTS: In the age groups of 55-64, 65-74 and 75-79 years, the incidence rate was 2.0, 5.8 and 17.3 per 1000 person years, and the prevalence 1.2, 4.2 and 8.0%, respectively. Definite or possible coronary heart disease was diagnosed in 46.0%, heart failure in further 20.2% and valvular heart disease or cardiomyopathy in 4.5%. In bivariate analysis adjusted for age, the following factors were significantly associated with future hospitalization for fibrillation: a family history of myocardial infarction, stroke in mother, dyspnoea at entry, alcohol abuse, high body stature and body weight, high blood pressure but not diabetes, high serum cholesterol, high heart rate, smoking, coffee consumption or psychological stress. Significant risk factors in multivariate analysis were age, odds ratio (OR) [95% confidence interval (CI)] 1.11 (1.07, 1.16) per year, hospitalization for coronary heart disease or heart failure -6.77 (5.17, 8.87), stroke in mother - 1.49 (1.15, 1.93), high body stature -1.04 (1.03, 1.06) per cm, high body mass index (BMI) -1.07 (1.04, 1.10) per kg m(-2), as well as hypertension -1.33 (1.07, 1.65). After a diagnosis of atrial fibrillation, mortality was increased by 3.3 times. CONCLUSION: In spite of a clinical association with coronary heart disease, risk factors for atrial fibrillation were only partly the same. Prevention includes avoidance of weight gain and control of blood pressure as well as prevention of myocardial infarction and heart failure. PMID- 11887973 TI - Asymptomatic versus symptomatic persistent atrial fibrillation: clinical and noninvasive characteristics. AB - OBJECTIVE: This prospective study was designed to investigate the differences between asymptomatic versus symptomatic arrhythmia as well as left ventricular dysfunction in a consecutive population of patients with persistent atrial fibrillation. DESIGN: A total of 282 consecutive outpatients referred with persistent atrial fibrillation formed the study population. A structured medical history was obtained. A two-dimensional transthoracic echocardiography to assess the left ventricular function and a 24-h electrocardiogram (ECG) recording were performed. Irregularity of the heart rhythm was analysed with heart rate variability (HRV) in the time domain as well as maximum and minimum heart rate and the longest pause. SETTING: Three university hospitals. RESULTS: The mean age of the patients was 69 years and the mean duration of atrial fibrillation was 7 months. The prevalence of symptomatic patients was 68%, while 32% had no symptoms from atrial fibrillation, left ventricular dysfunction was observed in 20%. Asymptomatic subjects had more often lone atrial fibrillation than those with symptoms. Valvular heart disease was an independent predictor of symptoms while male gender, ischaemic heart disease and a high heart rate were independent predictors of impaired left ventricular function. CONCLUSION: Valvular heart disease is related to symptoms in persistent atrial fibrillation. Ischaemic heart disease, male gender and a high heart rate are more common in patients with impaired left ventricular function. Compromised left ventricular function does, occur also in asymptomatic subjects underlining the importance of a careful investigation including echocardiography in all subjects with persistent atrial fibrillation. PMID- 11887974 TI - Episodes of ST-segment depression is related to changes in ambulatory blood pressure and heart rate in intermittent claudication. AB - OBJECTIVE: To study the prevalence and circadian distribution of ischaemic ST segment depression detected with ambulatory electrocardiographic monitoring (AECG) in patients with intermittent claudication (IC) as well as to study ambulatory blood pressure (ABP) and the relation of ischaemic episodes to variations in ABP and heart rate. DESIGN: A total of 40 patients with a history of IC and an ankle/brachial-index (ABI) <0.9 performed: (i) 24-h AECG recordings, (ii) simultaneous 24 h recordings of ABP every 15 min (Spacelabs 90207), (iii) an exercise treadmill test (ETT). An ischaemic episode was defined as a transient ischaemic ST-segment deviation > or =1 mm lasting >1 min. Eleven patients were excluded from ECG analysis because of uninterpretable ECG caused by treatment with digoxin or technical problems. RESULTS: Out of 29 patients, eight experienced a total of 15 episodes of ST-depression on AECG. The mean duration was 21+/-31 min. The majority of episodes (11 of 15) occurred between 6 and 12 a.m. In eight patients with ST-segment depression three had a history of ischaemic heart disease (IHD), four were hypertensives and four had signs of myocardial ischaemia on ETT. There were no significant differences between patients with and without ST-segment depression in ABP, walking performance or ABI. During ST-depression episodes systolic and diastolic blood pressure and heart rate were higher than day mean values; 178+/-41 vs. 166+/-30 mmHg (P= 0.09); 96+/-9 vs. 90+/-4 mmHg (P = 0.01) and 103+/-9 vs. 87+/-5 beats min(-1) (P < 0.01). CONCLUSION: Silent myocardial ischemia occurred in about a third of patients with IC. Episodes of ischaemia were associated with an increased ABP and heart rate. Whether treatment of high blood pressure may reduce silent ischaemia and if this favourably influences outcome is a matter of further research. PMID- 11887975 TI - Rosiglitazone treatment of patients with extreme insulin resistance and diabetes mellitus due to insulin receptor mutations has no effects on glucose and lipid metabolism. AB - BACKGROUND: Rosiglitazone, a thiazolidinedione (TZD), increases insulin sensitivity by reducing levels of plasma NEFA, triglycerides (TG), glucose and serum insulin. Rosiglitazone treatment decreases insulin resistance in type 2 diabetic patients, but no data exist concerning rosiglitazone treatment of patients with syndromes of extreme insulin resistance. OBJECTIVES: To evaluate whether hyperglycaemia in two lean patients with primary severe insulin resistance due to insulin receptor (IR) mutations and diabetes mellitus could be reduced by supplement of rosiglitazone for 180 days and secondary, to evaluate the effects on plasma NEFA, TG, Apo B, PAI-1 and serum insulin. SUBJECTS: Both patients (brothers) have known mutations in the IR gene localized to the tyrosine kinase domain and a deletion of exon 17 in part of their IR mRNA. Prior to the study the HbA1c values were higher than 10% in both patients for more than 12 months during treatment with insulin and metformin. RESULTS: After 180 days of rosiglitazone supplement (8 mg day(-1)), no changes were observed in fasting plasma glucose and HbA1c. Incremental plasma glucose areas under the curves during a 75-g oral glucose tolerance test (OGTT) were unchanged. Likewise, no improvements were seen in either first or second phase insulin secretion during a 0.3 g kg(-1) intravenous glucose tolerance test (IVGTT). Fasting plasma VLDL and HDL cholesterol, TG and Apo B levels were unchanged, whereas a small increase was seen in total and LDL cholesterol levels. Fasting plasma NEFA increased by 51% in KC after 90 days of treatment, and after 180 days plasma NEFA was still 26% higher, when compared with pretreatment levels. In BC an initial 16% decrease was seen in plasma NEFA after 90 days of treatment. Plasma NEFA was increased 14% after 180 days of treatment, when compared with pretreatment levels, but 35% when compared with day 90. Plasma PAI-1 decreased in both patients after 45 and 90 days of treatment but the decrease was only maintained in KC (47%). CONCLUSIONS: Rosiglitazone treatment, in combination with insulin and metformin, of patients with severe primary insulin resistance due to IR mutations and diabetes mellitus, had no impact on the measured estimates of glucose and lipid metabolism. These findings may suggest that the effect of rosiglitazone on glucose and lipid metabolism are dependent on the presence of intact IR protein. PMID- 11887976 TI - Evidence for the regulation of levels of plasma adhesion molecules by proinflammatory cytokines and their soluble receptors in type 1 diabetes. AB - OBJECTIVES: To investigate the regulation of soluble adhesion molecules by tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and relationships with circulating cytokine receptors, in vivo, in type 1 diabetes. DESIGN: Cross sectional study. SETTING: University hospital diabetes clinic. SUBJECTS: A total of 47 non-nephropathic, Caucasian type 1 diabetics and 39 nondiabetic controls. OUTCOME MEASURES: Plasma levels of TNF-alpha, IL-6, their soluble receptors and adhesion molecules intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1), sE-selectin and von Willebrand factor (vWF), and risk factors for cardiovascular disease. RESULTS: Plasma concentrations of IL-6 were elevated in diabetic patients compared with controls [median (interquartiles) 1.28 (0.89-2.65) vs. 0.66 (0.45-1.73) pg mL(-1): P=0.016], and in these patients IL-6 and soluble IL-6 receptor (sIL-6R) levels correlated with concentrations of sICAM-1 (r = 0.41, P = 0.012 and r = 0.31, P = 0.04, respectively). Tumour necrosis factor-alpha soluble receptor-2 (sTN-FRII), but not TNF-alpha or tumour necrosis factor soluble receptor-1 (sTNFRI), was elevated in diabetic subjects (P = 0.027). Plasma TNF-alpha levels correlated with sVCAM-1 (r = 0.39, P = 0.008), triglycerides (r = 0.36, P = 0.02 1) and diastolic blood pressure (r = 0.35; P = 0.024). Both sTNFRI and sTNFRII correlated with blood pressure (r = 0.46, P = 0.002; r = 0.32, P = 0.034) and triglycerides (r = 0.33, P = 0.033; r = 0.29, P = 0.05). In contrast, HDL-cholesterol and triglyceride were related to sE-selectin (r = -0.45 and +0.45; both P < 0.001). Neither sE selectin nor vWF were related to cytokine concentrations. Finally, both TNF-alpha and sIL-6R correlated sTNFRI and RII (r = 0.44-0.49, P < 0.001). None of these interactions were apparent in control subjects. CONCLUSIONS: (i) IL-6, through effects on sICAM-1, and TNF-alpha via effects on sVCAM-1, may promote vascular adhesion; (ii) plasma levels of TNF-alpha are associated with dyslipidaemia and increased blood pressure, adding to vascular disease risk; (iii) the actions of both cytokines are probably modified by altered production of soluble receptors in diabetic subjects. PMID- 11887977 TI - The yield of a diagnostic hospital dyspnoea clinic for the primary health care section. AB - OBJECTIVE: To investigate the impact of a combined examination programme with treatment advice on patients from general practice with dyspnoea. DESIGN: Prospective study with 6 months followup. SETTING: Regional hospital offering care for patients from 74 general practitioners. SUBJECTS: A total of 284 consecutive patients referred from general practice with dyspnoea. INTERVENTIONS: Patients were subjected to a combined examination programme including physical examination, ECG, chest X-ray, lung spirometry, echocardiography and routine laboratory tests. MAIN OUTCOME MEASURES: (i) Relationship between a diagnosis made by the referring general practitioner and the diagnosis based on the combined examination programme. (ii) The impact of the investigation programme and resulting therapeutic advice on dyspnoea after 6 months. RESULTS: Only in 39% of the patients there was concordance of the diagnoses on referral and the diagnosis based on the examination programme. Heart failure and lung disease was suspected in 126 and 79 patients, respectively, but these diagnoses were confirmed in only one-third to half of the patients. Conversely heart failure was revealed in 13 of 107 patients not suspected of heart failure (12%) and lung disease in 45 of 154 patients not suspected of pulmonary disease (29%). A change of treatment was suggested in 64% of all patients. After 6 months, improvement of dyspnoea was seen in more than half of the patients. In patients in whom the changes of medical treatment were completed, 61% expressed improvement in dyspnoea, whereas improvement of dyspnoea was recorded in only 34% of patients in whom the recommended treatment advice was not taken (P < 0.01). CONCLUSION: (i) In most patients it seems to be too difficult to establish the background of dyspnoea in general practice. (ii) There appears to be a substantial chance of improvement in patients with dyspnoea, in particular for patients who act on treatment advice based on an integrated examination programme; the chance of improvement is almost twice as good as in patients who are not capable to do so. PMID- 11887978 TI - Low risk of contrast-medium-induced nephropathy with modern angiographic technique. AB - OBJECTIVE AND DESIGN: A retrospective study of the incidence of contrast-medium induced nephropathy (CMN) in patients with renal insufficiency. SUBJECTS: All angiographies with and without endovascular therapeutic procedures (n = 2400) performed at the hospital during 1 year were evaluated. A total of 139 patients were found to have a preangiographic serum-creatinine (s-Cr) of 150 micromol L( 1) or above. Postprocedural serial s-Cr values were present in 118 patients and these were included in the study. RESULTS: Amongst patients receiving only iodinated contrast media (CM) 8% demonstrated a 25% rise in s-Cr. The corresponding figure was 11 and 12.5% amongst patients who were given either iodinated CM together with carbon dioxide (CO2) or CO2 as sole contrast medium. After exclusion of other explanations of impaired renal function all together only seven of 114 patients (6%) were considered to have developed CMN. Four of the seven patients restituted renal function completely, whilst it remained decreased in three. No patient required dialysis. The percentage of diabetic patients were not found to be different in patients with and without signs of CMN. CONCLUSIONS: The present retrospective study indicate that the risk of CMN in connection with angiography is low when modern low-osmolality CM and contrast saving angiographic technique including CO2 is used combined with proper hydration. Patients with diabetes mellitus were not found more frequently in the groups with CMN. PMID- 11887979 TI - Prediction of five-year survival for patients admitted to a department of internal medicine. AB - OBJECTIVE: The effect of many common forms of therapy, as medication for mild hypertension or hypercholesterolaemia, only reaches clinical significance after years of treatment. The meaningful application of such therapy presupposes that physicians can, at least to some extent, predict the remaining lifetime of patients. We investigated whether clinicians from different disciplines were able to predict the 5-year survival of patients admitted to a department of internal medicine. DESIGN: The members of two groups, each consisting of an internist, a surgeon and a general practitioner, made individual predictions of the expected remaining lifetime of discharged patients from written summaries of clinical information. Each patient was randomized to be assessed by the members of either of the two groups. The predictions were compared with actual 5-year survival. SETTING: Department of internal medicine at a university hospital. SUBJECTS: Patients admitted consecutively during a 6-week period. MAIN OUTCOME MEASURES: Sensitivity, specificity, positive and negative predictive values and areas under the receiver operating characteristic (ROC) curves for predictions of 5-year survival for each of the six experts. RESULTS: A total of 402 patients were included. Five-year survival was 0.63. The sensitivity of the predictions ranged from 0.81 to 0.95, the specificity from 0.61 to 0.77, the positive predictive value from 0.78 to 0.87 and the negative predictive value from 0.68 to 0.87. The areas under the ROC curves ranged from 0.84 to 0.91. CONCLUSION: The quality of predictions of 5-year survival made by experienced clinicians should permit the rational use of treatments with long-term effects. PMID- 11887980 TI - Effect of second and third generation oral contraceptives on lipid metabolism in the absence or presence of the factor V Leiden mutation. AB - OBJECTIVES: The effect of a second and third generation oral contraceptive and of the progestagens used in these pills on lipid metabolism was studied in the absence or presence of the factor V Leiden mutation. DESIGN: A single centre, double blind randomized trial. SETTING: University Medical Centre. SUBJECTS: A total of 51 women without and 35 women with the factor V Leiden mutation. INTERVENTIONS: A second generation (30 microg ethinylestradiol/150 microg levonorgestrel) or a third generation (30 microg ethinylestradiol/l 50 microg desogestrel) oral contraceptive. After two cycles of use and a wash-out period of two cycles, the participants received the corresponding progestagen-only preparation containing 150 microg levonorgestrel or 150 microg desogestrel. MAIN OUTCOME MEASURES: Mean difference in changes between the treatment groups on total cholesterol, HDL, LDL, triglycerides and total/HDL cholesterol ratio. RESULTS: Compared with levonorgestrel, desogestrel-containing oral contraceptives caused in women without the factor V Leiden mutation significant changes in HDL (0.43; 95% confidence interval [CI] 0.25-0.61), LDL (-0.55; 95% CI -0.90 to 0.20), triglycerides (0.19; 95% CI 0.06-0.32) and total/ HDL cholesterol ratio ( 0.87; 95% CI -1.21 to -0.53). When the progestagen-only preparations were used, differential changes were found for HDL (0.16; 95% CI 0.03-0.29), LDL (-0.31; 95% CI - 0.56 to -0.05) and total/HDL cholesterol ratio (-0.55; 95% CI -0.84 to 0.26). Desogestrel-only caused changes opposite to those of desogestrel containing oral contraceptives. For cholesterol and triglycerides, this effect was also found for levonorgestrel-only in comparison with levonorgestrel-combined oral contraceptives. Levonorgestrel appeared to induce the effect on HDL. Almost all results were similar for women with the factor V Leiden mutation. CONCLUSION: It appears that desogestrel counteracts the effects of oestrogens to a lesser extent than levonorgestrel. Desogestrel-containing oral contraceptives have therefore a more favourable influence on cholesterol metabolism in comparison with levonorgestrel-containing oral contraceptives. PMID- 11887981 TI - Two years of penicillin prophylaxis is sufficient to prevent clinically evident carditis in poststreptococcal reactive arthritis. PMID- 11887982 TI - Homocystinuria due to cystathionine beta-synthase deficiency associated with megaloblastic anaemia. PMID- 11887983 TI - Molecular mechanisms of prostate cancer. PMID- 11887985 TI - Radiotherapy and combined modality approaches in localised prostate cancer. PMID- 11887984 TI - Prostate cancer: natural history and surgical treatment of localised disease. AB - In summary, there is increasing and convincing evidence that radical prostatectomy is effective in locally confined, poorly differentiated prostate cancer. Diagnostic efforts, therefore, should be targeted toward this disease and probably also, based on the natural history evidence, toward moderately differentiated disease, mainly Gleason score 7. It is unclear, at present, how this can be achieved. Further improvement of our diagnostic capabilities is urgently needed. Hopefully ongoing randomised studies comparing radical prostatectomy to surveillance and studies comparing radical prostatectomy to radiotherapy are urgently desired. The randomised screening studies, which are ongoing, will provide important information with respect to the effect of treatment. If prostate cancer mortality in those men who are randomised to screening turns out to be better than in those randomised to control, this will also be an indication of the effectiveness of treatment. Also, the screening studies and associated natural history studies based on serum repositories and follow-up in non-screened patients will provide important information with respect to the natural history of prostate cancer in relation to PSA and changes of PSA over time. Finally, quality of life with and without treatment will have to be evaluated, in a prospective manner, in multicentre settings according to validated criteria such as those presented by Litwin. The outcomes of such studies will have to be added as utilities to data relating to traditional endpoints such as cancer-specific and overall survival. In the meantime, clinical practice will be determined by the fact that the only way to cure prostate cancer is early diagnosis and aggressive management. Encouragement comes from the increasing volume of evidence showing that poorly differentiated disease can be eradicated as long as it is locally confined. PMID- 11887986 TI - Systemic treatment and new developments in advanced prostate cancer. PMID- 11887987 TI - Biology and genetics of colorectal cancer. PMID- 11887989 TI - Palliative and adjuvant chemotherapy in colorectal cancer. PMID- 11887988 TI - Colorectal cancer--what is standard surgery? PMID- 11887990 TI - The response of the surgeon to changing patterns in breast cancer diagnosis. PMID- 11887993 TI - Epidemiology in Europe. PMID- 11887991 TI - The role of radiotherapy in rectal cancer. PMID- 11887994 TI - Importance of a comprehensive geriatric assessment in older cancer patients. PMID- 11887995 TI - Cancer surgery in the elderly. AB - Surprisingly, cancer in the elderly is frequently treated in a poor manner, and the behaviour of the disease in elderly patients is often poorly understood. Cancer treatment varies significantly with age, the percentage of patients receiving definitive treatment declines with increasing age, and there is a decline in survival of cancer with age. One of the contributing factors may be that physicians are less likely to recommend specialist consultation for elderly patients. One has to keep in mind that cancer surgery in the elderly, without any co-morbidity, is safe, and that, nowadays, the morbidity and mortality increases minimally with the age of the patient. In contrast, the morbidity and mortality is 2-3 fold as high in the elderly cancer patients with co-morbidity compared with younger cancer patients. A better public education of the elderly may increase cancer awareness, and therefore decrease the risk of developing symptoms that require emergency surgery, with a subsequent three times increase in the mortality rate. Surgeons treating elderly cancer patients should realise that performance status is more important than age, and should always keep in mind the three major questions as recently formulated by Balducci; (1) is the patient going to die of cancer or with cancer, (2) is the patient able to tolerate the surgery and possible surgical-related complications, and (3) is the patient likely to suffer the complications of cancer during her/his life? The increased number of surgical treatment options in elderly cancer patients will lead to an increase in overall cancer survival in elderly cancer patients, and contribute to an improvement in their quality of life. Surgical oncologists should focus on how to manage the most common cancers in elderly people such as breast, colorectal, lung and prostate, as well as take an active part in palliative treatments. PMID- 11887996 TI - Radiotherapy. PMID- 11887998 TI - Medical oncology. PMID- 11887997 TI - Pharmacology. PMID- 11887999 TI - Optimal comprehensive symptom control: an overall strategy. PMID- 11888001 TI - Present status of palliative radiotherapy. PMID- 11888002 TI - Nursing management in palliative care. PMID- 11888000 TI - The pharmacological management of cancer pain. PMID- 11888003 TI - Radiotherapy as part of a multidisciplinary treatment strategy in early breast cancer. PMID- 11888004 TI - Standard medical treatment for early breast cancer. PMID- 11888005 TI - What the clinician needs from the pathologist: evidence-based reporting in breast cancer. AB - Histopathology has a vital role in determining breast cancer management and pathologists must be part of the clinical team. Carcinoma size, grade, and especially lymph node status remain the best available prognostic factors. Metastatic carcinoma in axillary nodes is more important than any other prognostic factor presently available. ER status is an important predictor of response to endocrine manipulation, but its independent prognostic significance, and that of micrometastatic disease, circulating carcinoma cells and other molecular factors, even well-studied ones such as HER2 status, are less clear. Pathology is the first clinical speciality to subject its practice to rigorous scientific analysis, and it has stood up well. However, workers without appropriate experience in Pathology or scientific design have created difficulties by undertaking poorly planned studies with ill-defined end-points, lacking appropriate quality control. New analytical techniques and therapeutic targets make it essential that we learn from past mistakes and integrate pathologists into the research teams pursing clinical trials and the assessment of new bio-markers. Without this, input resource will be wasted on false leads that could have been curtailed. Morphology alone will not be enough to select patients likely to benefit in trials of new therapies, but selection 'tests' must be appropriate. The confusion of tests for selection of patients to receive Herceptin shows what happens when this process fails. Much of the microarray data being put into data-bases has no quality control, and meta-analysis of this data will produce even more conflict than the clinical trials. This can be avoided, as the ability to standardise is available. PMID- 11888006 TI - Chairman's introduction. Lung cancer. PMID- 11888007 TI - Molecular biology of lung cancer. PMID- 11888008 TI - Lung cancer: diagnosis and surgery. PMID- 11888009 TI - The role of radiotherapy and the value of combined treatment in lung cancer. PMID- 11888010 TI - State of the art in systemic treatment of lung cancer. PMID- 11888011 TI - Preliminary immunochemical test for the determination of ethyl glucuronide in serum and urine: comparison of screening method results with gas chromatography mass spectrometry. AB - Ethyl glucuronide is a highly specific metabolite of ethanol that is formed by enzymatic conjugation of ethanol with glucuronic acid. Because of its suitability as a marker of excessive alcohol consumption in serum with low blood-alcohol concentration and as a consumption marker in serum and urine, especially after the breakdown of ethanol, demand exists for a simple and fast analytical procedure, which is rarely possible using mass spectrometric determination methods. For this reason, we developed an immunochemical screening procedure (ELISA) in which polyclonal antibodies are bound to the walls of microtiter plates. To test suitability, 335 authentic serum and 186 urine samples were examined using immunochemistry and gas chromatography-mass spectrometry (GC-MS). The serum (urine) samples with cutoff values of 0.31 mg/L (1.33 mg/L) yielded false-negative results in 9.5% (24.3%) and false positives in 8.4% (23.2%) of cases. Specificity was calculated at 91.6% (76.8%) and sensitivity at 90.5% (75.7%). Test efficiency was 90.8% (76.3%). The study shows that ethyl glucuronide and therefore alcohol consumption can be detected in immunochemical screening of serum in a similar manner as current drugs, but the method is of limited value for urine. A GC-MS confirmation continues to remain a necessity. PMID- 11888012 TI - Dansyl chloride derivatization of methamphetamine: a method with advantages for screening and analysis of methamphetamine in urine. AB - The screening and quantitation of methamphetamine (MP) in urine using dansyl chloride (DNC) as the derivatization reagent were studied. Urinary MP derivatized with DNC could be detected by visual observation of the fluorescence in a solid phase extraction column such as a Sep-Pak C18 cartridge to which the whole reaction solution was applied. The DNC-derivatized MP was eluted from the cartridge and then identified and quantitated by gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). In the GC MS analysis with the MS detector in the electron-impact mode, DNC-derivatized MP and amphetamine (AP), exhibited diagnostic molecular ion peaks. The intensities of the molecular ions were 15% (DNC-MP) and 35% (DNC-AP) of the base peak (a fragment ion because of the loss of dimethylnaphthalene from M+), demonstrating that this method of derivatization has a major advantage for confirming APs by GC MS. MP derivatized with DNC could be determined by HPLC with ultraviolet detection. Because a good correlation (r = 0.95) between the GC-MS and HPLC method for urinary MP was confirmed, both HPLC and GC-MS appear to be useful tools for determining urinary MP. The intensity of the cartridge fluorescence due to DNC-derivatized MP was approximately related to the urinary content of MP determined by HPLC or GC-MS, although a false positive in the visual fluorescence was observed in some urinary specimens from healthy volunteers. From these results, screening and confirmation/determination following DNC derivatization is proposed as a suitable method for the analysis of MP. PMID- 11888013 TI - JAT's impact factor--room for improvement? Journal of Analytical Toxicology. AB - The impact factor of a scientific journal is simply the ratio of the number of citations to the number of citable items (articles and reviews) over a given time period, usually two years after the year of publication. Trends in the impact factor of Journal of Analytical Toxicology (JAT) are reviewed and compared with other leading journals in the forensic sciences and toxicology. In particular, the journals that frequently cite JAT articles (citing journals) and the journals cited in articles published in JAT (cited journals) are compared and contrasted. The reasons for citing a particular article are considered, and some suggestions are made for improving the impact factor of JAT, if this is deemed necessary. This could be achieved in a number of ways, such as speeding the editorial handling and peer-review processes, by including one or more invited review articles in each issue of the journal, or by increasing the number of references cited so the references/article ratio increases. Regardless of journal impact factor, an article should be judged by its usefulness to the field and not the prestige of the journal where it is published. PMID- 11888014 TI - Analysis of styrene and its metabolites in blood and urine of workers exposed to both styrene and acetone. AB - A purge-and-trap gas chromatographic (PT-GC) method for determining styrene concentrations in urine and blood samples has been used in the biological monitoring of workers exposed to styrene and acetone. Blood and urine samples were collected from 34 individuals exposed to both solvents at the end of a 4-h shift and measured for styrene in urine (Su), blood (Sb), and the two major urinary metabolites, mandelic acid (MA) and phenylglyoxylic acid (PGA). A second urine sample was taken at the beginning of the next shift. Environmental exposure was measured using passive personal monitoring and GC. Urinary excretion of MA and PGA was measured by high-performance liquid chromatography. The average exposures to styrene and acetone were 70.5 mg/m3 and 370.5 mg/m3, respectively. In end-of-shift samples there was a significant correlation between concentrations of Su and Sb and the metabolites PGA, MA (r = 0.714 and 0.788, p < 0.001 for Su and r = 0.644 and 0.566, p < 0.005 for Sb). A high correlation between Sb and Su (r = 0.732, p < 0.001) also existed. Poor correlations were found between Su and metabolites in samples collected at the beginning of the next shift (r = 0.491 and 0.474 for PGA and MA, respectively, p < 0.05). There was a better correlation between the biological parameters at the end of the shift and the environmental styrene (r = 0.841 for PGA, r = 0.834 for MA, r = 0.788 for Su, and r = 0.698 for Sb; p < 0.001) compared with those at the start of the shift (r = 0.81 for PGA, 0.675 for MA, and 0.650 for Su; p < 0.001). We found that the concentration of excreted metabolites decreased significantly when environmental concentrations of acetone increased (p < 0.05), particularly at the end of the shift. Although the best correlation with environmental styrene was obtained with the sum of PGA and MA at the end of the shift (r = 0.862, p < 0.001), urine and blood styrene were shown to be more useful biological monitoring indicators because their concentrations were not affected by acetone co-exposure. PMID- 11888015 TI - Endogenous nandrolone metabolites in human urine. Two-year monitoring of male professional soccer players. AB - 19-Norandrosterone (19-NA) and 19-noretiocholanolone (19-NE) are the two main indicators used to prove the illegal use of nandrolone by humans. Recent studies showed that 19-NA and 19-NE can be endogenously produced in some individuals. The mediated cases observed over the last three years generated some questions about the appropriateness of the official International Olympic Committee cutoff level, which is 2 ng/mL of 19-NA in male urine samples. In the present study, professional soccer players belonging to the French First League were studied over a period of 19 months. In total, 385 urine samples were taken immediately before and after soccer competitions and were coupled with 200 blood samples for testosterone and LH determination. Results of the study showed that the mean values for 19-NA and 19-NE were 0.097 ng/mL and 0.033 ng/mL, respectively. For 19 NA, 70% of the samples proved to be below 0.1 ng/mL, whereas less than 20% were found to be between 0.1 and 0.2 ng/mL, and 7% were between 0.2 and 0.3 ng/mL. Only four urine samples were above 1.0 ng/mL; the maximal value was 1.79 ng/mL. For 19-NE, only one sample was above 1.0 ng/mL; the value was 1.42 ng/mL. Concentrations of these compounds after games were generally significantly higher than those before games. PMID- 11888016 TI - Salting out improves solid-phase extraction recoveries on Abselut-Tox columns for broad spectrum drug screening. PMID- 11888017 TI - Distribution of triazolam and alpha-hydroxytriazolam in a fatal intoxication case. AB - The case of a 77-year-old woman who was found dead in her bathtub with her head clearly above the water line is presented. The decedent had a medical history of depression, liver disease, spinal stenosis, and diabetes mellitus. An empty medication bottle of triazolam was found in the trashcan. At autopsy, no injury or evidence of drowning was found. Toxicological analysis identified triazolam at a concentration of 0.12 mg/L in the heart blood. Triazolam and alpha hydroxytriazolam were quantitated in the specimens received. The medical examiner ruled that the cause of death was triazolam intoxication and the manner of death was suicide. PMID- 11888018 TI - A fatal human intoxication with the herbicide allyl alcohol (2-propen-1-ol). AB - Oral ingestion of allyl alcohol by a 55-year-old man resulted in death within 100 min. At autopsy, bloody, reddish fluid was found in mouth, larynx, esophagus, and trachea. The mucous membranes of the trachea, stomach, and duodenum were congested and inflamed. The stomach contained a pungent green-black fluid, and all internal organs exhibited a strong pungent odor. Toxicological analysis of blood identified allyl alcohol using solid-phase microextraction and gas chromatography-mass spectrometry. Quantitative determination of allyl alcohol and its toxic metabolite, acrolein, was performed using headspace gas chromatography with flame-ionization detection. Total amounts of allyl alcohol in gastric content, bile, and urine were 3.6 g, 15 mg, and 0.5 mg, respectively. The concentration in blood was 309 mg/L. Acrolein was not detected in gastric contents and only in small amounts in bile and urine. The concentration of acrolein in blood was 7.2 mg/L. Death was attributed to acrolein-induced acute cardiotoxicity, similar to that previously documented in animal experiments. PMID- 11888019 TI - High concentration of chloroquine in urine gives positive result with Amphetamine CEDIA reagent. PMID- 11888020 TI - Quantitation of organophosphorus nerve agent metabolites in human urine using isotope dilution gas chromatography-tandem mass spectrometry. AB - An isotope dilution gas chromatography-tandem mass spectrometric (GC-MS-MS) method was developed for quantitating the urinary metabolites of the organophosphorus nerve agents sarin, soman, tabun (GA), VX, and GF. Urine samples were concentrated by codistillation with acetonitrile, derivatized by methylation with diazomethane, and analyzed by GC-MS-MS. The limits of detection were less than 4 microg/L for all the analytes except for the GA metabolite, which had a limit of detection of less than 20 microg/L. PMID- 11888021 TI - Frederick Sanger--winner of 2 Nobel prizes. PMID- 11888022 TI - 113 letters from the Mayo Clinic: a pattern of medical referrals in the early 20th century. PMID- 11888023 TI - NSAIDs and a lower risk of prostate cancer: causation or confounding? PMID- 11888024 TI - A population-based study of daily nonsteroidal anti-inflammatory drug use and prostate cancer. AB - OBJECTIVE: To investigate the association between daily use of nonsteroidal anti inflammatory drugs (NSAIDs) and prostate cancer. PATIENTS AND METHODS: Subjects were 50- to 79-year-old white men randomly selected in January 1990 from the Olmsted County, Minnesota, community (n=1362) from among participants in a longitudinal study of lower urinary tract symptoms. At the beginning of the study, all medications that were used daily, including prescription and nonprescription NSAIDs, were ascertained by trained interviewers. Men who developed a histologically proved diagnosis of prostate cancer during a median of 66 months (maximum, 6 years) of follow-up were identified from a complete review of the community medical record. RESULTS: Twenty-three (4%) of 569 NSAID users and 68 (9%) of 793 nonusers developed prostate cancer during follow-up (P=.001). The relative odds of prostate cancer were 0.45 (95% confidence interval [CI], 0.28-0.73) in NSAID users compared with nonusers. This inverse association with NSAID use increased with increasing age at study onset. Thus, the relative odds of prostate cancer among NSAID users were 0.9 (95% CI, 0.4-2.2) in men aged 50 to 59 years, 0.4 (95% CI, 0.2-0.8) in men aged 60 to 69 years, and 0.2 (95% CI, 0.1 0.5) in men aged 70 to 79 years compared with similarly aged men who did not use NSAIDs. CONCLUSION: The study findings suggest that daily use of NSAIDs may be associated with a lower incidence of prostate cancer in men aged 60 years or older. The stronger effect among older men suggests that NSAIDs may prevent the progression of prostate cancer from latent to clinical disease. PMID- 11888025 TI - Role of transvenous implantable cardioverter-defibrillators in preventing sudden cardiac death in children, adolescents, and young adults. AB - OBJECTIVE: To evaluate the indications, underlying cardiac disorders, efficacy, and complications involved with transvenous implantable cardioverter defibrillators (ICDs) in pediatric patients at the Mayo Clinic. PATIENTS AND METHODS: The records of all patients aged 21 years or younger who underwent transvenous ICD placement at the Mayo Clinic, Rochester, Minn, were reviewed retrospectively. RESULTS: Between March 1992 and September 2000, 16 patients (7 females; mean age, 15.4 years; range, 10-21 years) underwent transvenous ICD placement. The ICD was implanted for primary prevention of sudden cardiac death in 7 and for secondary prevention in 9. The underlying cardiac disorders included hypertrophic cardiomyopathy in 6 patients and congenital long QT syndrome in 6 patients. The mean +/- SD follow-up was 36+/-29 months (range, 5-108 months). There was no mortality. Seven patients (44%) received appropriate ICD therapy, including 6 of 9 who had ICDs placed for secondary prevention. Median time free from appropriate ICD discharge was 3 years (range, 0.2-9 years). Three patients (19%) experienced inappropriate ICD discharge. Two patients needed device replacement because of technical problems (lead fracture and device malfunction). Two patients developed pocket infection that required removal and reimplantation of the ICD. CONCLUSION: In adolescents and young adults, transvenous ICDs may prevent sudden death but are not free of complications. Forty-four percent of this cohort received potentially life-saving ICD therapy, including two thirds who received an ICD for secondary prevention. PMID- 11888026 TI - Force assessment in periodic paralysis after electrical muscle stimulation. AB - OBJECTIVE: To obtain an objective measure of muscle force in periodic paralysis, we studied ankle dorsiflexion torque during induced paralytic attacks in hyperkalemic and hypokalemic patients. SUBJECTS, PATIENTS, AND METHODS: Dorsiflexor torque after peroneal nerve stimulation was recorded during provocative tests on 5 patients with hypokalemic or hyperkalemic disorders and on 2 control subjects (1995-2001). Manual strength assessment was simultaneously performed in a blinded fashion. Standardized provocation procedures were used. RESULTS: The loss of torque in hyperkalemic patients roughly paralleled the loss of clinically detectable strength, whereas in the hypokalemic patients, pronounced torque loss occurred well before observed clinical effects. No dramatic changes occurred in the control subjects. Torque amplitude decreased more than 70% in all patients during the provocation tests; such decreases were associated with alterations induced in serum potassium concentrations. CONCLUSIONS: Stimulated torque measurement offers several advantages in characterizing muscle dysfunction in periodic paralysis: (1) it is independent of patient effort; (2) it can show a definitely abnormal response early during provocative maneuvers; and (3) characteristics of muscle contraction can be measured that are unobservable during voluntary contraction. Stimulated torque measurements can characterize phenotypic muscle function in neuromuscular diseases. PMID- 11888027 TI - Association of parental vaccination reports with measles, mumps, and rubella protective antibody levels: comparison of Somali immigrant, Hispanic migrant, and US children in Rochester, Minn. AB - OBJECTIVES: To compare measles, mumps, and rubella antibody levels in Somali immigrant, Hispanic migrant, and US children in Rochester, Minn, and to determine whether parental vaccination reports predict seropositivity. SUBJECTS AND METHODS: From 1995 to 1997, we conducted a cross-sectional analysis using measles, mumps, and rubella antibody levels obtained from a sample of Somali, Hispanic, and Rochester children. Volunteers provided blood samples, vaccination histories, and demographic information. We assessed differences in measles, mumps, and rubella antibody levels among the 3 groups of children and calculated positive and negative predictive values to determine whether parental report of vaccination predicted seropositivity. RESULTS: Study participants included 79 Hispanic migrant, 69 Somali immigrant, and 730 Rochester children. Somali children reported vaccination at significantly older ages compared with Hispanic or Rochester children (P<.001). Most children were seropositive for all 3 antibodies. Parental reports of vaccination had high positive predictive values (71%-100%) but low negative predictive values (0%-50%). CONCLUSION: Somali and Hispanic children were as likely as Rochester children to be seropositive for measles, mumps, and rubella antibodies despite poor documentation of vaccination. Somali children, however, tended to receive vaccinations at significantly older ages than Hispanic and Rochester children. PMID- 11888028 TI - Evidence of an intrinsic sinus node abnormality in patients with postural tachycardia syndrome. AB - OBJECTIVE: To determine whether an intrinsic sinus node abnormality is involved in the pathophysiology of the postural tachycardia syndrome (POTS). SUBJECTS, PATIENTS, AND METHODS: In this prospective study, we compared the relationship between P-wave axis (PWA) and heart rate (HR) in 11 healthy controls and 14 patients with POTS by obtaining 12-lead electrocardiographic recordings during supine rest and during gradual head-up tilt. The HR of controls was titrated with isoproterenol infusion to match the HR of patients. The PWA was compared at different HR levels, and the relationship between HR and PWA was assessed for patients and controls. Primary end points were the PWA-HR relationship in healthy controls, comparison of these data with data from patients with POTS as a group, and identification of a possible subgroup of patients with POTS with irregular PWA-HR relationship. RESULTS: The PWA increased with increasing HR following a similar logarithmic trendline in both groups. The PWA of patients was significantly lower at the lowest comparable HR level but not different at faster HR levels. Three patients (21%) had a clearly abnormal HR-PWA relationship with substantial shift toward lower PWA. CONCLUSIONS: Our data support the hypothesis of a primary sinus node abnormality in a subset of patients with POTS. The ability to identify patients with primary sinus node abnormality may have important therapeutic implications. PMID- 11888029 TI - Diagnosis and treatment of migraine. AB - Despite recent advances in understanding the pathophysiology and treatment of migraine, considerable uncertainty remains surrounding the diagnosis and treatment of this disorder. This uncertainty is reflected in studies that show both underdiagnosis and undertreatment of migraine. While the diagnosis can be assisted by criteria from the International Headache Society, other approaches may be useful in clinical practice. Treatment of migraine must be based on an individualized patient strategy that integrates education, patient participation, and effective use of pharmacological interventions. Many patients, despite self treatment with simple analgesics, continue to suffer considerable disability associated with their migraines. Triptans, which are more effective at relieving migraine symptoms and maintaining patient function than are nonspecific therapies, are used in only a minority of patients with migraine. Treatment goals of rapid, complete relief with no recurrence and minimal adverse effects can be achieved when effective therapy is matched to individual patient goals. For prophylaxis, anticonvulsant drugs emerging as effective options are being added to the armamentarium with traditional compounds such as tricyclic antidepressants and beta-blockers. PMID- 11888030 TI - Upper gastrointestinal tract safety of risedronate: a pooled analysis of 9 clinical trials. AB - Risedronate sodium is a pyridinyl bisphosphonate effective for treatment and prevention of postmenopausal and glucocorticoid-induced osteoporosis. Some bisphosphonates have been associated with upper gastrointestinal (GI) tract adverse effects. The objective of this study was to determine the frequency of upper GI tract adverse events associated with risedronate, especially among high risk patients. The GI tract adverse events reported during 9 multicenter, randomized, double-blind, placebo-controlled studies of risedronate conducted from November 1993 to April 1998 were pooled and evaluated. The evaluation included 10,068 men and women who received placebo (n=5048) or 5 mg of risedronate sodium (n=5020) for up to 3 years (intent-to-treat population). Studies incorporated a comprehensive, prospective evaluation of GI tract adverse events. Adverse event information was collected every 3 months. The treatment groups were similar with respect to baseline GI tract disease and use of concomitant treatments during the studies. At study entry, 61.0% of patients had a history of GI tract disease and 38.7% had active GI tract disease; 20.5% used antisecretory drugs during the studies. Sixty-three percent used aspirin and/or nonsteroidal anti-inflammatory drugs (NSAIDs) during the studies. Upper GI tract adverse events were reported by 29.6% of patients in the placebo group compared with 29.8% in the risedronate group. The risk of experiencing such an event in the risedronate group was 1.01 (95% confidence interval, 0.94-1.09) relative to the placebo group (P=.77). The rate of upper GI tract adverse events per 100 patient-years was 19.2 in the placebo group compared with 20.0 in the risedronate group (P=.30). Risedronate-treated patients with active heartburn, esophagitis, other esophageal disorders, or peptic ulcer disease at study entry did not experience worsening of their underlying conditions or an increased frequency of upper GI tract adverse events overall. Concomitant use of NSAIDs, requirement for gastric antisecretory drugs, or the presence of active GI tract disease did not result in a higher frequency of upper GI tract adverse events in the risedronate treated patients compared with controls. Endoscopy, performed in 349 patients, demonstrated no statistically significant differences across treatment groups. The results of this extensive evaluation indicate that daily treatment with 5 mg of risedronate sodium is not associated with an increased frequency of adverse GI tract effects, even among patients at high risk for these events. PMID- 11888031 TI - A practical guide to the diagnosis and management of fecal incontinence. AB - Many physicians are unaware that fecal incontinence is often correctable. With appropriate and relevant diagnostic tests, medical treatment and/or surgical correction often leads to good functional results. General guidelines for diagnosis, evaluation, and management of fecal incontinence are provided. There are many causes of fecal incontinence, with obstetrical trauma being one of the most frequent. A detailed history, documentation of sphincter injury, and thorough physical examination will identify the cause of the problem in most patients. Management involves the use of antidiarrheal medication and fiber products, biofeedback, or enemas. A qualified surgeon should be consulted during the course of the patient's evaluation, particularly when medical therapy is unsuccessful. Knowledge of the appropriate diagnosis, evaluation, and management of fecal incontinence may result in more patients seeking medical attention and thus improving their quality of life. PMID- 11888032 TI - Evaluation and treatment of erectile dysfunction in men with diabetes mellitus. AB - Diabetic men have a more than 3-fold increased prevalence of erectile dysfunction (ED) compared with nondiabetic men. Erectile function is primarily a vascular phenomenon, triggered by neurologic controls and facilitated by appropriate hormonal and psychological components. Recent advances in the understanding of the physiology of penile vasculature and its role in male sexual performance have influenced the clinical approach to ED. The pathophysiological alterations leading to impotence in diabetic men include vasculogenic, neurogenic, and hormonal etiologies. A clinical work-up, including a thorough history and physical examination, is an important aspect of ED management. Biochemical evaluations to rule out secondary causes like hypogonadism and thyroid abnormalities are suggested. Oral medications acting through phosphodiesterase inhibition in penile vasculature have revolutionized treatment of impotence in diabetic men. Because of a high success rate in treating ED of various etiologies, these agents are the treatment of choice for most patients. Safety and efficacy of vacuum-constriction devices, intraurethral suppositories, intracavernosal injections, and other therapies are discussed. A clinical algorithm for the evaluation and management of ED is provided for use in the primary care setting. PMID- 11888033 TI - 58-year-old woman with abdominal pain. PMID- 11888034 TI - Hemophagocytosis in a patient with chronic lymphocytic leukemia and histoplasmosis. AB - We present a case of hemophagocytosis in the setting of a disseminated Histoplasma infection in a patient with B-cell chronic lymphocytic leukemia (CLL). A 68-year-old man with CLL presented with progressive pancytopenia and fevers after therapy with cyclophosphamide and fludarabine phosphate. Extensive evaluation for a source of infection revealed a pulmonary nodule. A biopsy specimen taken from the nodule showed granulomas containing Histoplasma organisms. A bone marrow biopsy specimen demonstrated disseminated histoplasmosis and intense hemophagocytosis. Antifungal therapy with amphotericin B was initiated, and the fevers and cytopenias resolved. Hemophagocytic syndrome is an uncommon condition with many origins. It is characterized by a proliferation of histiocytes with phagocytosis of formed elements of blood. Clinical manifestations include signs and symptoms of immune activation and decreased peripheral blood cell counts. This condition is often underdiagnosed because clinicians are unfamiliar with it. PMID- 11888035 TI - Reversible myelopathy in a 34-year-old man with vitamin B12 deficiency. AB - Vitamin B12 deficiency is common, with most patients lacking classic features of advanced severe deficiency. Early diagnosis and treatment prevent severe anemia and irreversible damage to the nervous system. We describe a 34-year-old man with pernicious anemia who presented with clinical and radiologic features of early myelopathy and borderline low serum levels of vitamin B12. Prompt diagnosis based on the measurement of serum methylmalonic acid and treatment with cyanocobalamin injections led to rapid resolution of clinical manifestations and magnetic resonance imaging abnormalities. We review the literature of magnetic resonance imaging in vitamin B12 deficiency myelopathy and discuss the issues relating to diagnosis and early treatment of this potentially reversible condition. PMID- 11888036 TI - Vitamin E use in preventing coronary heart disease in patients undergoing dialysis. PMID- 11888037 TI - Treatment of diastolic heart failure. PMID- 11888038 TI - Chest pain in patients with normal findings on angiography. PMID- 11888039 TI - Hidden danger of generic medications. PMID- 11888041 TI - Reveille for Radicals! The paramount purpose of health education? PMID- 11888040 TI - Octreotide treatment of acromegaly during pregnancy. PMID- 11888042 TI - Project Northland: long-term outcomes of community action to reduce adolescent alcohol use. AB - Project Northland was a randomized trial to reduce alcohol use among adolescents in 24 school districts in northeastern Minnesota. Phase 1 (1991-1994), when the targeted cohort was in grades 6-8, included school curricula, parent involvement, peer leadership and community task forces. The Interim Phase (1994-1996) involved minimal intervention. Phase 2 (1996-1998), when the cohort was in grades 11 and 12, included a classroom curriculum, parent education, print media, youth development and community organizing. Outcomes of these interventions were assessed by annual student surveys from 1991 to 1998, alcohol purchase attempts by young-looking buyers in 1991, 1994 and 1998, and parent telephone surveys in 1996 and 1998. Growth curve analysis was used to examine the student survey data over time. Project Northland was most successful when the students were young adolescents. The lack of intervention in the Interim Phase when the students were in grades 9 and 10 had a significant and negative impact on alcohol use. The intervention used with the high school students as those in grades 11 and 12 made a positive impact on their tendency to use alcohol use, binge drinking and ability to obtain alcohol. There was no impact in Phase 2 on other student-level behavioral and psychosocial factors. Developmentally appropriate, multi component, community-wide programs throughout adolescence appear to be needed to reduce alcohol use. PMID- 11888043 TI - Factors that enable and inhibit transition from a weight management program: a qualitative study. AB - Failure of clients to initiate closure and move out of weight management programs after it is considered they should have reached the stage of maintenance in their health behavior change has implications for clients' self-management, provision of health promotion programs and their cost-effectiveness. This study aims to identify factors that enable and inhibit class attendees' transition from a weight management program. Six short-term attendees who had left the program after two terms and six long-term attendees who had attended four or more terms of the program volunteered to participate in in-depth interviews. Enabling factors were identified to be program knowledge and attainment of set goal weight, and inhibiting factors were the perceived need to come to classes, concern about keeping in control, recognition of the potential to lapse and being involved with a group. Recommendations are made for the program to include a component addressing relapse prevention training and to trial some form of follow up support strategy. Additionally, further research is needed into transition from weight management programs. PMID- 11888045 TI - Ethnic differences in social correlates of diet. AB - Little is known about whether culture influences social correlates of dietary behaviors. Questionnaires on parent- and child-reported family and peer influences on children's fruit, juice and vegetable consumption were analyzed for ethnic group differences in responses. Grade 4-6 students completed the questionnaires in the classroom and their parents completed telephone or in-home interviews. Analyses of variance across ethnic categories and chi2 analysis of differences in ethnic group composition between clusters of scales were conducted. Few ethnic group differences were detected, suggesting substantial commonality among respondents. Ethnic differences might be accommodated by interventions tailored to particular behaviors among ethnic groups. PMID- 11888044 TI - Implementation of a teacher-delivered sex education programme: obstacles and facilitating factors. AB - Interventions are unlikely to achieve their desired aims unless they are implemented as intended. This paper focuses on factors that impeded or facilitated the implementation of a specially designed sex education programme, SHARE, which 13 Scottish schools were allocated to deliver in a randomized trial. Drawing on qualitative and quantitative data provided by teachers, we describe how this intervention was not fully implemented by all teachers or in all schools. Fidelity to the programme was aided by intensive teacher training, compatibility with existing Personal and Social Education (PSE) provision, and senior management support. It was hindered by competition for curriculum time, brevity of lessons, low priority accorded to PSE by senior management, particularly in relation to timetabling, and teachers' limited experience and ability in use of role-play. The nature of the adoption process, staff absence and turnover, theoretical understanding of the package, and commitment to the research were also factors influencing the extent of implementation across and within schools. The lessons learned may be useful for those involved in designing and/or implementing other teacher-delivered school-based health promotion initiatives. PMID- 11888046 TI - Testicular self-examination (TSE) among Dutch young men aged 15-19: determinants of the intention to practice TSE. AB - The present study analyzed what determinants are important to describe and explain the intention of testicular self-examination (TSE) for young men aged 15 19 attending senior high school (response rate 80%, n = 274). The questionnaire assessed determinants, including knowledge, attitude (positive and negative consequences, anticipated regret, and moral obligation), social influence (social norm, social support and modeling) and self-efficacy. Knowledge of testicular cancer and TSE was very low. Only 2% of the subjects reported regularly performing TSE. After hearing of TSE (through the questionnaire), 41% of all young men had a positive intention to start performing TSE regularly. The various intention groups (positive, neutral and negative) differed significantly on almost all of the determinants. Multiple regression analysis showed that young men who where anxious about TSE and those who were not anxious had different determinants explaining the variance in the intention to perform TSE regularly (R2 = 41-57%). Differences in determinants of intention between young men who are anxious about TSE and young men who are not can be used to design health education interventions that may therefore be more effective for these different subgroups. PMID- 11888047 TI - Reductions in smoking prevalence and cigarette consumption associated with mass media campaigns. AB - This paper examines reductions in smoking prevalence and cigarette consumption associated with state and local mass-media campaigns. We review the findings of the empirical literature on campaigns targeted at the general population. We then discuss the findings on state- and community-level youth-oriented campaigns. The results suggest that well-funded and implemented mass-media campaigns targeted at the general population and implemented at the state level, in conjunction with a comprehensive tobacco control program, are associated with reduced smoking rates among both adults and youth. Studies of youth-oriented interventions specifically have shown more mixed results, particularly for smaller, community-level media programs, but they indicate strong potential to influence underage smoking rates. We conclude by examining issues that warrant additional research. The scale and duration of expenditures, the content of ad messages, and other tobacco control polices are aspects of media programs that may help explain differences among study results. In particular, tobacco control polices that are implemented during the campaign often make it difficult to identify the specific influence of media campaigns alone. PMID- 11888048 TI - Evaluation of a nurse-managed minimal-contact smoking cessation intervention for cardiac inpatients. AB - This study examined the effectiveness of a nurse-managed minimal-contact smoking cessation intervention for patients hospitalized for cardiac disease. A pre-test post-test quasi-experimental design was used. Patients who smoked prior to admission to cardiac wards of five hospitals (n = 388) received the intervention, whereas smoking patients in six other hospitals were given usual care (n = 401). The intervention was initiated at the hospital and continued after discharge. The core elements were stop-smoking advice from the cardiologist, a short bedside consultation with a nurse, administration of self-help materials and aftercare by the cardiologist. Smoking cessation was assessed after 3 months by self-report. Logistic regression analysis excluding dropouts, controlling for covariates including baseline differences showed significant intervention effects (one tailed significance test) on point prevalence abstinence (OR = 2.11) and continuous abstinence (OR = 1.41). Intention-to-treat analysis including dropouts as smokers showed a significant effect on point prevalence abstinence (OR 1.35). We conclude that, compared to usual care, the low-intensity smoking cessation intervention for cardiac inpatients was more effective in achieving smoking cessation. However, the small effects and the process evaluation suggest that improvements are needed. PMID- 11888049 TI - Quantification of ephedrines in urine by column-switching high-performance liquid chromatography. AB - A method for the quantification of five congener ephedrines in urine samples without sample preparation was developed. The analytes were trapped on a C18 precolumn and separated on a C18 BDS analytical column. Baseline separation was achieved for all analytes. The method meets the requirements of the International Olympic Committee (IOC) medical commission regarding cut-off limits for positive doping cases with ephedrines. PMID- 11888050 TI - Determination of the anticancer drug KW-2170, a pyrazoloacridone derivative, and its metabolites in human and dog plasma by high-performance liquid chromatography using an electrochemical detector. AB - KW-2170, 5-(3-aminopropyl) amino-7,10-dihydroxy-2-(2-hydroxethyl)-aminoethyl-6H pyrazolo [4,5,1-de] acridin-6-one dihydroxychloride, is a novel anticancer agent under clinical development. We have established a highly sensitive method which can simultaneously quantitate KW-2170 and its two metabolites, a carboxylic (M1) and hydroxylated (M2) derivative involving the 5-position, in human and dog plasma. KW-2170 and its metabolites were extracted from plasma using a weak cation-exchange cartridge and then determined by HPLC using an electrochemical detector (ED). Over the concentration range 0.1-50 ng/ml, precision and accuracy of intra- and inter-day assay were within 11% in human plasma. In dog plasma, they were within 17% at the lower quantitation limit and within 11% at other concentrations. These three compounds were stable during the assay procedure, freeze-thawing cycles and during long-term storage. Using this methodology, the pharmacokinetics of KW-2170 in a dog could be monitored over 24 h. This method is suitable for evaluation of the detailed pharmacokinetics of KW-2170 and its metabolites in humans and dogs. PMID- 11888051 TI - Normal-phase and stability-indicating reversed-phase high-performance liquid chromatographic methods for the determination of the novel antitumor agent: 1 methylpropyl-2-imidazolyldisulfide. AB - 1-Methylpropyl-2-imidazolyl disulfide (MID) is a novel antitumor agent currently in Phase I clinical trials. The chromatographic behavior of MID and its potential impurity, degradation product, and metabolite 2-mercaptoimidazole (2MI) was studied under reversed-phase (RP) and normal-phase (NP) conditions. Both RP- and NP-HPLC separation methods were developed. RP-HPLC was validated as a stability indicating assay for MID. NP-HPLC retained both MID and 2MI and pending further validation, could prove useful in the study of MID pharmacokinetics. PMID- 11888052 TI - Method for integrated analysis of polycyclic aromatic hydrocarbons and organochlorine compounds in fish liver. AB - An analytical method for integrated analysis of organochlorine compounds and polycyclic aromatic hydrocarbons (PAH) in large numbers of fish liver samples has been developed using one single clean-up step. Tissues are homogenized with anhydrous sodium sulphate and Soxhlet extracted with n-hexane-dichloromethane (4:1, v/v) for 24 h. The extracts are cleaned-up and fractionated with an alumina chromatographic column allowing the separation of the extracts in two fractions. One containing most organochlorine compounds, including hexachlorobenzene, DDTs and polychlorobiphenyls, and the other the hexachlorocyclohexane isomers and PAH. These two fractions are subsequently analysed by GC-MS. Tests of repeatability result in relative standard deviations mainly under 20%. Evaluation by the standard addition method shows good linearities and recoveries. PMID- 11888053 TI - Simple and sensitive method for the determination of celecoxib in human serum by high-performance liquid chromatography with fluorescence detection. AB - A simple method is described for the determination of the cyclooxygenase-2 specific inhibitor celecoxib in human serum by HPLC using the demethylated analogue as internal standard. After protein precipitation with acetonitrile, samples were extracted with chloroform. Separation was achieved on a Prontosil C18 AQ column (150x3 mm I.D., 3-microm particle size) at a flow-rate of 0.35 ml/min using water-acetonitrile (40:60, v/v) as the mobile phase. Using fluorescence detection with excitation at 240 nm and emission at 380 nm, the limit of quantification was 12.5 ng/ml for a sample size of 0.5 ml of serum. The assay was linear in the concentration range of 12.5-1500 ng/ml and showed good accuracy and reproducibility. At all concentrations intra- and inter-assay variabilities were below 11% with less than 9% error. The method was applied to the determination of celecoxib for pharmacokinetic studies in man. PMID- 11888054 TI - Determination of donepezil, an acetylcholinesterase inhibitor, in human plasma by high-performance liquid chromatography with ultraviolet absorbance detection. AB - A simple and sensitive high-performance liquid chromatographic (HPLC) method with UV absorbance detection is described for the quantification of donepezil, a centrally and selectively acting acetyleholinesterase inhibitor, in human plasma. After sample alkalinization with 0.5 ml of NaOH (0.1 M), the test compound was extracted from I ml of plasma using isopropanol-hexane (3:97, v/v). The organic phase was back-extracted with 75 microl of HCl (0.1 M) and 50 microl of the acid solution was injected into a C18 STR ODS-II analytical column (5 microm, 150x4.6 mm I.D.). The mobile phase consisted of phosphate buffer (0.02 M, pH 4.6), perchloric acid (6 M) and acetonitrile (59.5:0.5:40, v/v) and was delivered at a flow-rate of 1.0 ml/min at 40 degrees C. The peak was detected using a UV detector set at 315 nm, and the total time for a chromatographic separation was approximately 8 min. The method was validated for the concentration range 3-90 ng/ml. Mean recoveries were 89-98%. Intra- and inter-day relative standard deviations were less than 7.3 and 7.6%, respectively, at the concentrations ranging from 3 to 90 ng/ml. The method shows good specificity with respect to commonly prescribed psychotropic drugs, and it could be successfully applied for pharmacokinetic studies and therapeutic drug monitoring. PMID- 11888055 TI - Sensitive and specific liquid chromatographic-tandem mass spectrometric assay for dihydroergotamine and its major metabolite in human plasma. AB - A sensitive and specific procedure for the simultaneous determination of dihydroergotamine (DHE) and its 8'-hydroxylated metabolite (8'-OH-DHE) in human plasma was developed and validated. The analytes were extracted from plasma samples by liquid-liquid extraction, separated through a Zorbax C18 column (50x2.1 mm I.D.) and detected by tandem mass spectrometry with an electrospray ionization interface. Caroverine was used as the internal standard. The method has a lower limit of quantitation (LOQ) of 10.0 and 11.0 pg/ml for DHE and 8'-OH DHE, respectively. The intra- and inter-run precision was measured to be below 9.1% for both DHE and 8'-OH-DHE. The inter-run accuracy was within 4% for the analytes. The overall extraction recoveries of DHE and 8'-OH-DHE were determined to be about 58 and 52% on average, respectively. The chromatographic run time was approximately 2.5 min. More than 120 samples could be assayed daily with this method, including sample preparation, data acquisition and processing. The method developed was successfully used to investigate plasma concentrations of DHE and 8'-OH-DHE in a pharmacokinetic study of volunteers who received DHE orally. PMID- 11888056 TI - Simultaneous assay of sildenafil and desmethylsildenafil in human plasma using liquid chromatography-tandem mass spectrometry on silica column with aqueous organic mobile phase. AB - A liquid chromatography-tandem mass spectrometry method was developed for the analysis of sildenafil (SIL) and its metabolite desmethylsildenafil (DMS) in human plasma. Samples were accurately transferred to 96-well plates using a liquid handler (Multiprobe II). Solid-phase extraction was carried out on a 96 channel programmable liquid handling workstation (Quadra 96) using a C8 and cation-exchange mixed-mode sorbent. The extract was injected onto a silica column with an aqueous-organic mobile phase, a combination that was novel for improving the method sensitivity. The low limit of quantitation was 1.0 ng/ml for both SIL and DMS. The method was validated to meet the criteria of current industrial guidance for quantitative bioanalytical methods. PMID- 11888057 TI - Hyphenation of sedimentation field flow fractionation with flow cytometry. AB - Interest in the development of field flow fractionation (FFF) systems for cell sorting recently increased with the possibility of collecting and characterizing viable cellular materials. There are various tools for the analysis of cell characteristics, but the reference is small- and large-angle light scattering often coupled with fluorimetric measurements. The well-known flow cytometry (FC) cell analysis techniques can be associated with FFF leading to the possibility of collecting information provided by a remarkable separation technique for micron sized particles (cells) operating in the steric-hyperlayer elution mode with multiparametric detection provided by flow cytometry. Moreover FFF derived cell characteristics can be correlated with FC characteristics to describe in a unique way the nature of the eluted materials. Experimental demonstrations are described herein using nucleated cells (HL-60 cell lineage) and human red blood cells (HRBC). PMID- 11888058 TI - Determination of urinary and salivary cotinine using gas and liquid chromatography and enzyme-linked immunosorbent assay. AB - The objective of this study was to compare cotinine concentrations in urine and saliva using gas chromatography (GC), high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA). Ninety-four subjects were selected (27 smokers and 67 non-smokers) and interviewed using questionnaire. Of the non-smokers, 39 had been exposed to environmental tobacco smoke (ETS) and 28 had not been exposed to ETS. Cotinine levels among smokers were highest using all three measurements, followed by ETS exposed subjects and non-smokers. Cotinine levels in urine, using HPLC, correlated significantly with levels measured using ELISA (r=0.92) and GC-nitrogen-phosphorus detection (NPD) (r=0.92). Salivary cotinine levels measured using ELISA did not correlate significantly with either HPLC (r=0.37) or GC-NPD (r=0.33) measurements. Multiple regression models were used to adjust for age, gender, drug use and health status, and it was found that cotinine levels in urine and saliva were significantly correlated with smoking pack-year. The authors conclude that urinary cotinine concentration is a more accurate biomarker for ETS than salivary cotinine concentration. PMID- 11888059 TI - Determination of the benztropine analog AHN-1055, a dopamine uptake inhibitor, in rat plasma and brain by high-performance liquid chromatography with ultraviolet absorbance detection. AB - N-Substituted 3alpha-[bis(4'-fluorophenyl)methoxy] tropanes represent a series of novel potential cocaine abuse therapeutics. AHN-1055, a member of this series, has been assessed to be the most suitable analog for pharmacokinetic studies. A sensitive and specific high-performance liquid chromatography method was developed to quantitate AHN-1055 in rat plasma and brain tissue. Reversed-phase chromatography with ultraviolet detection (lambda=220 nm) was utilized to quantitate the eluate. Plasma or brain tissue samples were prepared by liquid liquid extraction using hexane, followed by evaporation, reconstitution in mobile phase, and injection onto an ABZ+plus column. AHN-1055 and oxprenolol (internal standard) eluted at approximately 9.9 and 5.01 min, respectively, without any interfering peaks. The calibration curves were found to be linear in the range of 25-10000 ng/ml for plasma and 50-5000 ng/g for brain (r2> or =0.999). The intra- and inter-day variabilities were < or =10% whereas the intra- and inter-day errors were < or =8.5%. Plasma and brain recoveries of AHN-1055 were 95 and 79%, respectively. Stability studies showed plasma quality control samples to be stable through at least three freeze-thaw cycles (error<3.5%), for at least 24 h when subjected to room temperature (error<3%) and for at least 30 h after loading the processed samples onto the autosampler (error<3%). AHN-1055 stock solution was found to be stable for at least 4 months when stored at 4 degrees C (error<6%). The validated method accurately quantified AHN-1055 in plasma and brain samples collected from a pharmacokinetic study consisting of an intravenous bolus in the tail vein of adult male Sprague-Dawley rats. PMID- 11888060 TI - Quantification of terbutaline in urine by enzyme-linked immunosorbent assay and capillary electrophoresis after oral and inhaled administrations. AB - The International Olympic Committee and World AntiDoping Agency restricts the use of beta2-agonists and only the inhaled administration of terbutaline, salbutamol, formoterol and salmeterol is permitted for therapeutic reasons. The aim of this study was to develop a test for the quantitation of terbutaline in urine and evaluate different parameters to distinguish between oral and inhaled administration of the drug. Urine samples were collected from asthmatic and non asthmatic recreational swimmers who had received repeated doses of oral (3x2.5 mg plus 1x5 mg during 24 h) and inhaled (12x0.5 mg in 24 h with half of it being in the last 4 h) racemic terbutaline, and single oral (5 mg) or single inhaled doses (1 mg). Total terbutaline concentrations (free+conjugated) were determined by enzyme-linked immunosorbent assay. Results showed that after oral administrations urinary terbutaline concentrations were higher than those detected after inhalation. For confirmation purposes, a chiral capillary electrophoretic procedure was established and validated. A solid-phase extraction with Bond-Elut Certify cartridges was undertaken, separation performed using a 50 mM phosphate buffer (pH 2.5) containing 10 mM of (2-hydroxypropyl)-beta-cyclodextrin as running buffer and diode-array UV detection set at 204 nm. The proposed procedure is rapid, selective and sensitive allowing quantitation of free terbutaline enantiomers in urine. No statistical differences were found between total free terbutaline concentrations [S-(+)+R-(-)] in urine collected after oral and inhaled administrations of the drug. After oral doses enantiomeric [S-(+)]/[R-( )] ratios lower than those obtained after inhalation were observed probably due to an enantioselective metabolism that take place in the intestine, but differences between both routes of administration were not statistically significant. Although different trends were observed after oral and inhaled doses in total terbutaline, total free terbutaline concentrations and in ratios between its enantiomers, differences observed were not sufficiently significant to establish cut-off values to clearly distinguish between both routes of administration. PMID- 11888061 TI - High-throughput quantification of the anti-leukemia drug STI571 (Gleevec) and its main metabolite (CGP 74588) in human plasma using liquid chromatography-tandem mass spectrometry. AB - The signal transduction inhibitor STI571 (formerly known as CGP 57148B) or Gleevec received fast track approval by the US Food and Drug Administration (FDA) for treatment of chronic myeloid leukemia (CML). STI571 is a revolutionary and promising new oral therapy for CML, which functions at the molecular level with high specificity. The dramatic improvement in efficacy compared to existing treatments prompted an equally profound increase in the pace of development of Gleevec. The duration from first dose in man to completion of the New Drug Application (NDA) filing was approximately 2.6 years. In order to support all pharmacokinetics studies with sufficient speed to meet various target dates, a semi-automated procedure using protein precipitation was developed and validated. A Tomtec Quadra 96 (Model 320) and a protein precipitation step in a 96-well plate format were utilized. A Sciex API 3000 triple quadrupole mass spectrometer with an atmospheric pressure chemical ionization interface operated in positive ion mode was used for detection. The method proved to be rugged and allowed the simultaneous quantification of STI571 and its main metabolite (CGP 74588) in human plasma. Herein, assay development, validation, and representative concentration-time profiles obtained from clinical studies are presented. PMID- 11888062 TI - Quantification of methyldopa in human plasma by high-performance liquid chromatography-electrospray tandem mass spectrometry application to a bioequivalence study. AB - A method based on LC-MS-MS is described for the determination of methyldopa in human plasma using dopa-phenyl-D3 as the internal standard. The method has a chromatographic run time of 5.5 min and was linear in the range of 20-5000 ng/ml. The limit of quantitation was 20 ng/ml, the intra-day precisions were 7.3, 5.4 and 4.3% and the intra-day accuracies were -8.0, -1.3 and -2.0% for 30, 600 and 3000 ng/ml, respectively. The inter-day precisions were 7.7, 0.5 and 0.7% and the inter-day accuracies were 0.2, -1.1 and -2.3%, respectively, for the above concentrations. This method was employed in a bioequivalence study of two tablet formulations of methyldopa. PMID- 11888063 TI - Analytical method for monitoring concentrations of cyclosporin and lovastatin in vitro in an everted rat intestinal sac absorption model. AB - Cyclosporin A (CSA) and lovastatin (LV) are lipophilic drugs, which show poor and erratic absorption when administered perorally. The permeability of these compounds can be increased transiently by altering the membrane characteristics of the absorptive epithelium by the use of sorption promoters (SPs). In the present work a simple validated HPLC method utilizing an isocratic mobile phase with short retention times for CSA and LV was developed in order to monitor their concentrations in Kreb's Ringer bicarbonate (KRB) solution in vitro in intestinal sac absorption model. The same method was utilized to determine the apparent permeability coefficients and absorption profiles of CSA and LV by a modified Wilson-Wiseman method. Drugs were analysed by a reversed-phase HPLC method using a Shim-pack C18 column. An isocratic mobile phase containing acetonitrile and water in the proportions 70:30 and 80:20 was used for the HPLC analysis of CSA and LV, respectively. The flow-rate was 2 ml/min and quantitative determinations were carried out at 215 nm at 70 degrees C for CSA. In the case of LV the flow rate was 1 ml/min and detection was done at 238 nm at 25 degrees C. The method was found to be specific as none of the proposed SPs, components of KRB or intestinal sac artefacts interfered with the drug peaks. Recovery studies and intra- and inter-day variations were within statistical limits. The limits of detection were 250 and 10 ng/ml and the limits of quantitation were 400 and 30 ng/ml for CSA and LV, respectively. The calibration curve was found to be linear in concentration range of 0.5-6 microg/ml for CSA and 0.05-0.4 microg/ml for LV. The proposed method was found to be rapid and selective and hence can be applied for continuous monitoring of CSA and LV in vitro in intestinal sac absorption studies. PMID- 11888064 TI - Relationship between lipophilicity and antitumor activity of molecule library of Mannich ketones determined by high-performance liquid chromatography, clogP calculation and cytotoxicity test. AB - A series of Mannich ketones were synthesized in order to study the relative importance of structure and specific substitutions in relation to their lipophilicity and antitumor activity. Substitutions were carried out with morpholinyl, pirrolidinyl, piperidyl and tetrahydro-isoquinolyl groups in various positions on three different skeletons. Lipophilicity of Mannich ketones was characterised by chromatography data (log k') and by software calculated parameters (clogP). Compounds were tested on their ability to inhibit the proliferation of the A431 human adenocarcinoma cell line evaluated by MTT and apoptosis assays. The results suggest that the higher the lipophilicity values (log k' and clogP), the higher the antitumor and apoptotic activity of Mannich ketones. Determination of lipophilicity by measuring the log k' or by calculating the clogP values of the compounds may help to predict their biological activities. PMID- 11888065 TI - Quantification of cholesterol in foods using non-aqueous capillary electrophoresis. AB - A simple method for the rapid quantification of cholesterol in egg yolk and milk by non-aqueous capillary electrophoresis (NACE) is described in this paper. The samples were treated with saponification and then quantified by NACE, in which 100 mM sodium acetate-acetic acid in methanol was employed as the running buffer. The correlation coefficient between the cholesterol concentration and the corresponding peak area was 0.999. The detection limit of cholesterol was 5 microg/ml (twice the signal-to-noise ratio). This method can be used as a routine method for the rapid and sensitive determination of cholesterol in foods. PMID- 11888066 TI - Pneumocystography in nonpalpable breast cysts: effect on remission rate. AB - The purpose of the study was to evaluate the effect on remission rate after pneumocystography among nonpalpable cysts. A series of 206 nonpalpable cysts aspirated using the perforated compression plate technique was reviewed. The effect on remission was evaluated on mammograms obtained 1-3 years after the cyst aspiration. Logistic regression was used to compare the effect between those examined with pneumocystography (n=62) and those aspirated alone (n=144). The ratio of complete remission was 52% (32/62) with pneumocystography compared to 53% (76/144) without. In univariate analysis there was no association between pneumocystography and complete remission. However, complete emptying of the cyst was significantly associated with complete remission (OR = 1.85, 95%CI = 1.05 3.25). In a multivariate model, complete emptying without pneumocystography was significantly associated with complete remission (OR = 2.40, 95%CI = 1.14-5.02) but not complete emptying in combination with pneumocystography (OR = 0.84, 95%CI = 0.24-2.89). Pneumocystography showed a close to two-fold association with complete remission. However, this association was not statistically significant (OR = 1.92, 95%CI = 0.52-7.05). In conclusion, complete emptying of a nonpalpable cyst significantly increased the chance of complete remission when pneumocystography was not performed. Pneumocystography showed no significant effect on remission rate. PMID- 11888067 TI - Inhibition of carbonic anhydrase in the lens does not induce myopia in cynomolgus monkeys. AB - Refraction was measured in eyes of cynomolgus (Macaca irus) monkeys, before and during continuous intravenous infusion of large doses of the carbonic anhydrase (CA) inhibitors acetazolamide and ethoxzolamide. No changes of refraction were seen. Therefore, inhibition of CA in lens, cornea and retina does not appear to be the cause of the transient myopia and associated symptoms, occasionally observed in patients treated with CA inhibitors. PMID- 11888068 TI - Microsurgical decompression without laminectomy in lumbar spinal stenosis. AB - Our objectives were to study a/ the clinical results of microsurgical decompression without laminectomy compared to those reported from standard decompression laminectomy in patients with central lumbar spinal stenosis, and b/ if the microsurgical technique could prevent post-operative instability and concomitant symptoms. Twenty-one patients were treated, 11 men and 10 women, aged 47-81 years. Fourteen patients had "pure" stenosis whereas 7 had additional diseases that compounded the symptoms of stenosis. Independent examiners saw the patients pre-operatively and a mean of 27 months post-operatively. Plain X-ray films were taken of 14 patients a mean of 5 years post-operatively to study possible slippage. Among the 14 patients with "pure" stenosis the results were excellent in 13 and fair in 1. Among the 7 with additional diseases the outcome was excellent in 1, fair in 1, unchanged in 3 and worse in 2. The technique did not prevent post-operative slippage, which occurred in 3 of 14 patients. However, the clinical outcome was not related to slippage. We found the microsurgical technique safe and gentle with excellent possibilities for decompression of the complete spinal canal without laminectomy. The results following this procedure were well comparable to or even better than those reported following standard decompression laminectomy. PMID- 11888069 TI - Informal parental traffic education and children's bicycling behaviour. AB - The aim of this study was to analyze the relation between traffic intensity and traffic hazards in the local traffic environment, the parents' view of their child's traffic situation and the actions taken by the parents to cope with these hazards. 58 parents were interviewed. The traffic intensity in the vicinity of the home was estimated. 19% of the parents lived in inner city areas, 62% in suburbs and 19% in the countryside. Suburban children had a safe traffic environment. Inner city- and countryside children predominately lived in high intensity traffic environment. Inner city- and suburb children frequently used the bike, in inner city as a tool for play and in the suburbs mainly as a means of transport. Countryside children seldom used their bicycle. In inner city areas carefulness in traffic and in the other two independence was emphasized. Only 16% reported cooperation between home and school on traffic matters. Traffic accidents were concentrated to children living in suburbs (p<0.01). 81% of accidents were reported by parents having independence as a goal for traffic training (p<0.01). The results underline that traffic accident risk is strongly contingent on the local traffic environment and informal parenteral education in traffic safety. PMID- 11888070 TI - Nerve cells in transplanted pancreatic islets: no effects of cyclosporin or tacrolimus on immediate neuronal survival. AB - Previous experiments have demonstrated that neuronal cells within pancreatic islets survive the isolation procedure and constitute an integral part of transplanted pancreatic islets. The aim of the present study was to investigate to what extent immunosuppressive drugs affects the acute survival of intra-islet neurons after pancreatic islet transplantation. For this purpose, C57BL/6 mice were syngeneically transplanted with 250 islets under the renal capsule. The animals were treated for 7 consecutive days with subcutaneous injections of cyclosporin, tacrolimus or vehicle. After this, the animals were killed and the grafts were removed, fixed and stained for the presence of the neuron-specific protein PGP 9.5. The number of nerves were then morphologically quantitated. No differences between the experimental groups were seen, and the number of nervous elements were approximately 5 per mm2 in all animals. It is concluded that immunosuppressive treatment does not affect the acute survival of graft neurons after experimental islet transplantation. PMID- 11888071 TI - Failure of loop diuretics to improve the long term outcome of ischaemic damage in rat kidneys. AB - The effects of furosemide administered at the onset of postischaemic renal failure were investigated in Sprague-Dawley rats one month after exposing the left kidney to 45 min of renal ischemia. In the experimental group, 13 mg furosemide was given intravenously both before and a few minutes after induction of the ischaemia and then, by an osmotic pump, in a daily dose of 2-3 mg for the following 7 days. The animals of the control group were treated similarly but with saline alone. After one month, the glomerular filtration rate (GFR) in the damaged left kidneys of the furosemide-treated rats was 0.5+/-0.08 ml/min, which was not significantly different from that in the untreated control rats, of 0.8+/ 0.14 ml/min. As expected, the right intact kidneys responded with an increase in GFR to about 2 ml/min. Further effects that were similar in the damaged kidneys of the furosemide-treated and untreated animals were a decrease in potassium secretion and in the urine concentration ability; the urine osmolality in the diseased left kidneys was thus 1000-1500 mOsm/kg, as against over 2000 mOsm/kg in the right, intact kidneys. The function of the individual nephrons in terms of such variables as single nephron filtration rate, fractional fluid reabsorption and tubular and vascular hydrostatic pressures remained unaltered, however. Hence, the severe reduction in whole kidney GFR appeared to be due to a loss of nephrons rather than to an equal decrease in each individual nephron. It is also clear that furosemide did not improve the long-term outcome of acute postischaemic renal failure. PMID- 11888072 TI - Age-related changes in neuroendocrine system of the gut. A possible role in the pathogenesis of gastrointestinal disorders in the elderly. Minireview based on a doctoral thesis. PMID- 11888073 TI - Extraction of IgY from egg yolk using a novel aqueous two-phase system and comparison with other extraction methods. AB - Egg yolk is an important source of antibodies. The biggest obstacle for isolation of chicken antibodies (IgY) is the removal of lipids, which are present in abundance in egg yolk. We have used a two-phase system to separate egg yolk. The use of an aqueous two-phase system with phosphate and Triton X-100 made separation of lipids and water-soluble proteins possible. Lipids are extracted into the detergent-enriched top-phase, whereas IgY is isolated in the phosphate enriched bottom-phase. The phosphate:triton system was characterised and optimised using various experimental designs. For the optimised model, the yield of IgY was kept above 97% (11.1-14.9 mg IgY/g egg yolk recovered). The amount of lipids in the bottom-phase was kept below 25% of the total content in the egg yolk added. Hence, the model described provides a method for extracting the IgY fraction with a high yield and relatively low lipid content. PMID- 11888074 TI - The advent of targeted therapeutics and implications for pathologists. PMID- 11888075 TI - ST1571 (imatinib mesylate) reduces bone marrow cellularity and normalizes morphologic features irrespective of cytogenetic response. AB - The tyrosine kinase inhibitor STI571 (imatinib mesylate, Gleevec) is an effective treatment for chronic myeloid leukemia (CML). We examined bone marrow samples from 53 patients with CML who were receiving STI571 in 3 multicenter phase 2 trials to assess morphologic changes and cytogenetic response to this drug. In most patients with initially increased blasts, the bone marrow blast count rapidly decreased during STI571 therapy. Reductions in cellularity, the myeloid/erythroid ratio (commonly with relative erythroid hyperplasia), and reticulin fibrosis (if present pretreatment) also were seen in most patients, resulting in an appearance resembling normal marrow in many cases. Eighteen patients (34%) had some degree of cytogenetic response. Surprisingly, these striking morphologic changes occurred irrespective of any cytogenetic response to STI571. Thus, STI571 seems to affect the differentiation of CML cells in vivo, causing even extensively Philadelphia chromosome-positive hematopoiesis to exhibitfeatures resembling normal hematopoiesis. PMID- 11888076 TI - Epstein-Barr virus-associated B-cell lymphoproliferative disorders in angloimmunoblastic T-cell lymphoma and peripheral T-cell lymphoma, unspecified. AB - Various patterns of Epstein-Barr virus (EBV)-associated B-cell lymphoproliferation occur in patients with immunodeficiency. We studied 17 cases of T-cell lymphoma displaying extensive EBV-driven B-cell lymphoproliferation or simultaneous/subsequent EBV-associated B-cell lymphoma. In 10 cases of angioimmunoblastic T-cell lymphoma, an uncommonly prominent population of EBV+ atypical, activated, focally confluent large transformed B cells was found in the background of T-cell lymphoma. In 4 cases, an EBV-associated B-cell neoplasm (3 diffuse large B-cell lymphomas, 1 plasmacytoma) occurred in patients with T-cell lymphoma. Three cases were composite lymphomas of a peripheral T-cell lymphoma, unspecified, combined with EBV-associated diffuse large B-cell lymphoma. The transformed B-cell population displayed EBV latency types 2 and 3. Monoclonal and oligoclonal B-cell populations were detected in 5 and 6 cases, respectively. Similar to other states of immunodeficiency, disease-related and therapy-induced immunosuppression in T-cell lymphoma may lead to a prominent EBV-associated B cell lymphoproliferation and to EBV+ B-cell neoplasms. PMID- 11888078 TI - Carisoprodol: an unrecognized drug of abuse. AB - During a 6-month monitoring period, carisoprodol was detected in the urine specimens of 19 patients for whom drug screening had been ordered for purposes of patient care. The clinical history suggested that in 7 cases the drug was abused or implicated in a suicide attempt or gesture. In another 7 cases, the drug was used primarily for medical purposes, and in 5 cases the reason for use could not be determined. One patient ingested homemade tablets that were found to contain carisoprodol. In an additional case, the drug was detected in breast milk. Physical findings, clinical history, and treatment are described, and the profile of a typical carisoprodol user is discussed. It seems that carisoprodol has become an unrecognized drug of abuse, at least in our community. This drug and its metabolite, meprobamate, should be included in comprehensive drug screening. PMID- 11888077 TI - The immunophenotype of 325 adult acute leukemias: relationship to morphologic and molecular classification and proposal for a minimal screening program highly predictive for lineage discrimination. AB - Bone marrow cells of 325 adults with acute leukemia were immunophenotyped using a panel of monoclonal antibodies proposed by the European Group for the Immunological Characterization of Leukemias (EGIL). Of these, 97.2% could be assigned clearly to myeloid or lymphoid lineage (254 acute myeloid leukemias [AMLs], 48 B-cell lineage acute lymphoblastic leukemias [ALLs], 14 T-cell lineage ALLs), 1.8% as biphenotypic, and less than 1% as undifferentiated. Immunologic subtyping of ALLs revealed an association between early precursor phenotypes and coexpression of myeloid antigens, particularly CD15/CD65s coexpression and pre pre-B cell-specific phenotypes and genotypes. The common ALL phenotype was associated with BCR-ABL translocation. Among AMLs, CD2 coexpression was almost exclusively restricted to French-American-British subtypes M3 variant and M4Eo and related molecular aberrations. The most valuable markers to differentiate between myeloperoxidase-negative AML subtypes M0 and ALLs were CD13, CD33, and CD117, typical of M0, and intracytoplasmic CD79a, intracytoplasmic CD3, CD10, and CD2, typical of B cell- or T cell-lineage ALL. Our results confirm excellent practicability of the EGIL proposalfor immunologic classification of acute leukemias. For myeloperoxidase-negative AMLs, we suggest a scoring system based on markers most valuable to distinguish between AML-M0 and ALLs. PMID- 11888079 TI - Automated sperm concentration analysis with a new flow cytometry-based device, S FCM. AB - The S-FCM uses flow cytometry technology to measure sperm concentrations. Semen samples from 104 men attending a male infertility clinic were used to evaluate the reproducibility of results and the carryover rate with the S-FCM by performing between- and within-run imprecision analyses. In addition, sperm concentrations measured with the S-FCM were compared with those obtained by manual analyses with the Makler chamber and the improved Neubauer hemacytometer. The results showed that automated analyses with the S-FCM were highly reproducible and the carryover rate was 0.17% or less. In within-run imprecision assays, the coefficients of variation for the S-FCM were less than 5% at all sperm concentrations, while those for the Makler chamber were between 17.7% and 28.7% at lower sperm concentrations. The overall correlation between values measured with the S-FCM and those measured with the Makler chamber and improved Neubauer hemacytometer was excellent, but at lower sperm concentrations the correlation was lower. The S-FCM performed sperm concentration analyses in 110 seconds compared with 5 minutes for the Makler chamber and 10 minutes for the improved Neubauer hemacytometer. The S-FCM is suitable for quantitative measurement of lower sperm concentrations. PMID- 11888080 TI - To step or not to step: an approach to clinically diagnosed polyps with no initial pathologic finding. AB - We determined whether there were additional diagnostic findings in additional level sections performed on polyps with no pathologic diagnosis (NPD) or those in which only lymphoid aggregates (LAs) were seen initially and determined the level at which findings were identified. All colorectal biopsy specimens submitted with a clinical diagnosis of polyp during a 6-month period were included (N = 733). Initially, 3 level sections were cut for each polyp, and if a cause for the polyp was found, no additional levels were evaluated. If LAs or no cause for the polyp was found, 5 additional levels through each block were examined. Any diagnostic findings and the level at which they were identified were recorded. A discrete cause for the polyp was identified in routine levels in 574 cases (78.3%). Deeper levels were performed in 159: 23 for clarification of a suspected diagnosis, 38 for LAs, and 98 for NPD. Findings were identified in 31 (22.8%) of 136 stepped for LA or NPD with neoplastic findings in 13 (9.6%). Most diagnoses were identified in levels 4 or 5, but tubular adenomas were found in levels 7 and 8. These results support level sectioning specimens submitted as polyps with NPD or LAs on initial sections. PMID- 11888081 TI - Deeper examination of negative colorectal biopsies. AB - Initial histologic sections of specimens from colorectal biopsies of putative lesions may lack polyps. These sections may contain lymphoid aggregates that seemingly correlate with endoscopic findings; however; additional sections might contain polyps. We reviewed 83 specimens from colorectal biopsies of putative lesions for which initial sections lacked polyps. Our objectives were to determine the incidence of polyps within additional sections and to determine whether the presence of lymphoid aggregates within initial sections excludes the presence of polyps within additional sections. Eight specimens (10%) contained polyps (5 adenomatous, 3 hyperplastic), which remained histologically occult until examination to depths of approximately 120 to 380 microm. Five polyps (62%) were associated with lymphoid aggregates that were present within initial sections. We conclude that additional sections may contain surprisingly large numbers of polyps and that lymphoid aggregates present within initial sections fail to exclude the presence of polyps within additional sections. PMID- 11888083 TI - Can technology expedite the cervical cancer screening process? A Hong Kong experience using the AutoPap primary screening system with location-guided screening capability. AB - We studied the usefulness of an automated screening instrument for processing Papanicolaou (Pap) smears to determine whether it could speed human examination by recording the time to screen 1,007 cervical Pap smears with an AutoPap primary screening instrument with location-guided screening (LGS) software and by conventional microscopic examination. We also assessed the accuracy of the methods to determine preparation adequacy, and we compared the diagnosis by each method. The AutoPap with LGS satisfactorily determined the adequacy of Pap smears and identified the marked abnormal cells for human examination. An accurate diagnosis was possible when only the marked cells were examined, and this method reduced the screening time to less than half that required for conventional screening. With low-grade squamous intraepithelial and more severe lesions as a threshold, there were 37 cases in the conventional group and 29 cases in the LGS group. With atypical squamous cells of undetermined significance as the threshold, there were 111 cases in the LGS group and 93 cases in the conventional group. The AutoPap with LGS can significantly speed the examination of Pap smears without lowering the detection rate of clinically important lesions, thus helping alleviate the cytotechnologist shortage. PMID- 11888082 TI - Failure to detect human papillomavirus DNA in malignant epithelial neoplasms of conjunctiva by polymerase chain reaction. AB - To elucidate the putative role of human papillomavirus (HPV) infection in the etiology of conjunctival tumors, 44 formalin-fixed, paraffin-embedded specimens of conjunctival tumors (24 patients with papillomas and 20 patients with dysplastic and/or malignant tumors) were screened for HPV infection using 4 different polymerase chain reactions (PCRs). Of the 24 samples of papilloma, 14 (58%) displayed positive results by applying nested PCR using primer sets of HPV consensus L1 region. HPV type 6 or 11 was detected in 9 cases of papilloma by type-specific primer sets, but none of them were positive for HPV type 16 or 18. However, by using the highly sensitive PCR technique, we failed to demonstrate the HPV DNA of HPV types 6, 11, 16, and 18 in any of the 20 malignant epithelial tumors of conjunctiva. We conclude that HPV-6 or HPV-11 is present in a substantial percentage of conjunctival papillomas, which is in accordance with findings of previously reported studies. In contrast, malignant conjunctival carcinomas are not associated with HPV infection; other pathogenic mechanisms, such as UV light, probably are more important in the cause of these malignant lesions. PMID- 11888084 TI - Transitional cell carcinoma of the renal pelvis the diagnostic role of pelvic washings. AB - One hundred renal pelvic washings were reviewed blindly for 12 cytologic features. Of 52 washings with tissue confirmation, the cytologic diagnosis suggestive of or positive for transitional cell carcinoma (TCC) was made in 36 cases; 11 were negative, and 5 were unsatisfactory. Of 36 positive washings, histology confirmed the TCC diagnosis in 35 but revealed only reactive changes in 1. Of 11 negative washings, 9 were histologically negative for TCC, and 2 were positive for high-grade TCC. Among 48 washings without tissue confirmation, 33 were negative for TCC or showed reactive changes, 12 were negative for high-grade dysplasia or malignancy, but low-grade TCC could not be ruled out, 1 was suggestive of malignancy, and 2 were unsatisfactory. Clinical follow-up revealed no TCC. Predictive cytologic features of high-grade TCC were high nuclear/cytoplasmic ratio, isolated cells, anisonucleosis, nuclear hyperchromasia, and coarse chromatin; for low-grade they were presence of more than 5 papillary groups, cellular overlapping, anisonucleosis, and hyperchromasia. The sensitivity and specificity for the cytologic diagnosis were 89% and 97% for high-grade TCC and 100% and 78% for low-grade TCC, respectively. Renal pelvic washings can be used to accurately diagnose TCC of the renal pelvis. The positive predictive value for high-grade TCC is 93%, but for low-grade tumors it is 43%. PMID- 11888085 TI - Beta-catenin expression and its association with prognostic factors in adenocarcinoma developed in Barrett esophagus. AB - The majority of the adenocarcinomas arising in Barrett esophagus manifest clinically at an advanced stage and have a poor prognosis. As a result of this poor prognosis, much attention has been directed toward the exploration of markers for neoplastic progression in Barrett esophagus. The objective of the present study was to determine the expression of beta-catenin by immunohistochemical analysis in 70 adenocarcinomas developed in Barrett esophagus and to examine its relationship to various prognostic factors currently in use. Abnormal beta-catenin expression, consisting of the loss of membranous staining and the appearance of the nuclear staining, was found in 43 cases (61%). Of patients with the 43 tumors showing abnormal beta-catenin expression, 25 (58%) survived more than 1 year. In contrast, only 7 (26%) of 27 patients with tumors showing normal beta-catenin expression survived longer than 1 year. Most of the superficial (Tis-T1) tumors (83% [10/12]) exhibited abnormal beta-catenin expression compared with only 53% (31/58) in the T2-T3 group. These results suggest a possible correlation among beta-catenin expression, tumor stage, and length of survival as prognostic factors in patients with adenocarcinoma in Barrett esophagus. PMID- 11888086 TI - The unsatisfactory ThinPrep Pap Test: missed opportunity for disease detection? AB - Cervical cytology specimens classified as "unsatisfactory for interpretation" represent a potential source of undetected disease. This prospective study analyzed the potential benefits of a laboratory procedure to reprocess unsatisfactory ThinPrep Pap Tests (Cytyc, Boxborough, MA). All unsatisfactory ThinPrep samples were reprocessed using a glacial acetic acid wash. The study period unsatisfactory rate was compared with that for the previous 12 months. The initial unsatisfactory rate was 1.3% (197/15,154). Of the unsatisfactory ThinPrep samples, 55.8% (110/197) had residual material for reprocessing. After reprocessing, 67.3% (74/110) were reclassified as "satisfactory" or "satisfactory but limited by," and the final unsatisfactory rate was 0.8% (123/15,154), a 62% decrease. Compared with the previous 12-month rate of 0.9% (209/23,730), this was a 12% reduction. Seven (6.4%) of 110 initially classified as unsatisfactory contained an epithelial abnormality (atypical squamous cells of undetermined significance, 3; atypical glandular cells of undetermined significance, 2; low grade squamous intraepithelial lesion, 1; squamous cell carcinoma, 1) on the reprocessed slide. Reprocessing of unsatisfactory ThinPrep slides yielded additional cellular abnormalities that otherwise would have been undetected. The present study confirms that reprocessing of unsatisfactory ThinPrep slides is a beneficial laboratory procedure. PMID- 11888087 TI - Lung adenocarcinoma with bronchioloalveolar carcinoma component is frequently associated with foci of high-grade atypical adenomatous hyperplasia. AB - We assessed the occurrence of atypical adenomatous hyperplasia (AAH) in whole lung lobes with primary cancer lesions. Following surgical resection, tissue specimens were sliced to a thickness of 4 mm (3,641 specimens from 61 cases; mean = 59.7 specimens per case). A total of 119 AAH foci were found and an association was evident in 25 (57%) of 44 adenocarcinomas, 3 (30%) of 10 squamous cell carcinomas, and 2 (29%) of 7 other lung cancers. Histologic evaluation showed that 108 AAH foci were categorized as low-grade and the other 11 as high-grade AAH. These 11 foci of high-grade AAH were present in 7 patients with adenocarcinoma, and in 1 patient there was a synchronous double primary lung adenocarcinoma. High-grade AAH was closely associated with bronchioloalveolar carcinoma (BAC) type adenocarcinoma, and low-grade AAH with non-BAC adenocarcinoma. The mean +/- SD Ki-67 labeling index in high-grade AAH (3.5%+/ 2.9%) was significantly higher than for the low-grade index (1.4%+/-1.6%). We propose that foci of high- but not low-grade AAH may be potential precursor lesions of lung adenocarcinoma, especially with the BAC component. PMID- 11888088 TI - Immunophenotypic characterization of 225 prostate adenocarcinomas with intermediate or high Gleason scores. AB - This study provides detailed staining results for 225 prostate adenocarcinomas, including 150 Gleason score 8, 9, and 10 adenocarcinomas with cytokeratins (CKs) 7, 20, 5/6, and 17, prostate-specific antigen (PSA), prostatic acid phosphatase (PAP), carcinoembryonic antigen (CEA), WT1, thyroid transcription factor-1 (TTF 1), and villin. CK7 was reactive in 112 adenocarcinomas (49.8%). The percentage of CK7-reactive adenocarcinomas and the percentage of CK7-stained cells increased in higher Gleason score adenocarcinomas; most reactive neoplasms had CK7 staining of fewer than 25% of cells. CK20 had similar results. The percentage of PSA- and PAP-reactive adenocarcinomas and the percentage of stained cells in reactive neoplasms decreased in higher Gleason score adenocarcinomas. CK5/6 and CK17, WT1, CA-125, TTF-1, and villin were nonreactive. The prostate can be the primary site of metastatic adenocarcinoma that is nonreactive for PAP and PSA and has CK7 or CK20 reactivity in fewer than 50% of the cells. The likelihood that a metastatic adenocarcinoma is from the prostate is low if reactivity with any of the cytokeratin antibodies, CEA, TTF-1, CA-125, WT1, or villin is extensive. PMID- 11888089 TI - Immunocytochemical localization of mast cells in lymphocytic hypophysitis. AB - We studied 15 transsphenoidally resected pituitary tissues diagnosed by histologic examination as chronic lymphocytic hypophysitis. Six autopsy-obtained pituitaries of patients who died of nonendocrine diseases also were studied. Tryptase immunohistochemical analysis, which specifically identifies mast cells, demonstrated numerous, randomly distributed multifunctional cells throughout the inflammatory reaction. Several mast cells were located in the vicinity of capillaries; several others were distributed far from the blood vessels. Occasional mast cells also were noted in the nonpathologic anterior and posterior pituitary lobes. Morphometric analysis confirmed that in lymphocytic hypophysitis, the number of mast cells per volume of tissue was significantly increased compared with that of nonpathologic anterior and posterior pituitary lobes. To elucidate the possible role of mast cells in chronic lymphocytic hypophysitis, microvessel densities were assessed quantitatively using immunohistochemical analysis for CD34, a sensitive marker of endothelial cells. The strong positive correlation between numeric density of mast cells and microvessel density per volume of pituitary tissue suggests that mast cell derived products may influence capillary permeability and angiogenesis, thereby facilitating the access of inflammatory cells to adenohypophysial cells. PMID- 11888090 TI - Nephroblastoma: multidrug-resistance P-glycoprotein expression in tumor cells and intratumoral capillary endothelial cells. AB - The development of chemoresistance in a variety of cancers seems related to overexpression of the P-glycoprotein (P-gp) drug pump. Nephroblastoma, the most common malignant renal tumor of childhood, usually is responsive to treatment, and prognosis is favorable in most cases. However, the disease in a subset of patients is refractory to treatment, and the disease follows an aggressive course. To study P-gp expression in this tumor and its correlation with outcome, tumor samples from 93 patients were examined by immunohistochemical analysis. P gp expression was determined separately in both tumor cells and intratumoral capillary endothelium. The likelihood ratio test, the Kaplan-Meier method, and the log-rank test were used to evaluate its association with clinical course, grade, stage, and administration of preoperative chemotherapy. The results for the majority of nephroblastomas were variably positive; in 43 (46%) of them, newly formed capillary endothelial cells also stained positive. While no association of P-gp expression in tumor cells with clinical course, stage, and grade could be demonstrated, positivity in endothelial cells correlated significantly with unfavorable outcome, suggesting that chemoresistance depended on an active blood-tumor barrier. Previous chemotherapy induced P-gp overexpression in tumor cells. PMID- 11888091 TI - ROC curves for thyroid FNA and HER-2/neu in breast cancer. PMID- 11888092 TI - Differentiation of bacterial from viral infections by a simple single test slide: the leukergy test. PMID- 11888093 TI - Waldenstrom macroglobulinemia caused by extranodal marginal zone B-cell lymphoma. PMID- 11888094 TI - Evaluation of serum creatine kinase in ectopic pregnancy with reference to tubal status and histopathology. AB - OBJECTIVE: To study the value of creatine kinase in ectopic pregnancy with reference to tubal histopathology. DESIGN: Prospective controlled study. SETTING: Academic tertiary-care institution. POPULATION: Thirty-two women with ectopic pregnancy and 20 controls with intrauterine pregnancies. METHODS: Creatine kinase and beta-hCG levels were measured on admission. Ectopic pregnancies were removed at surgery and examined histologically. MAIN OUTCOME MEASURES: Tubal localisation and integrity of ectopic pregnancies as judged at surgery and later histologically, and placental growth patterns in unruptured ectopic pregnancies classified as intraluminal, extraluminal or mixed as determined histologically. RESULTS: Creatine kinase levels were higher in isthmic than ampullary ectopic pregnancies (P = 0.011), and higher in ruptured than in unruptured cases (P = 0.003) and normal pregnancies (P < 0.0001). A creatine kinase value >120iu/L was 65% sensitive and 87% specific in discriminating ruptured from unruptured ectopic pregnancies. Creatine kinase levels were above this cutoff in two of five unruptured ampullary ectopic pregnancies with invasive trophoblastic growth, yet in none of nine cases with intraluminally confined placentation (P = 0.04). Creatine kinase was positively correlated with gestational age in ruptured (P = 0.007), but not in unruptured ectopic pregnancies or normal pregnancies. CONCLUSIONS: Serum creatine kinase may help in discriminating ruptured from unruptured ectopic pregnancies, while it is not useful for the primary diagnosis of ectopic pregnancy. An increase in creatine kinase levels accompanying muscular damage in ectopic pregnancy probably antedates tubal rupture, and may be related to trophoblastic growth patterns. PMID- 11888095 TI - Intrauterine 10 microg and 20 microg levonorgestrel systems in postmenopausal women receiving oral oestrogen replacement therapy: clinical, endometrial and metabolic response. AB - OBJECTIVE: The clinical and endometrial efficacy and lipid response of two different doses of intrauterine levonorgestrel were assessed in comparison with sequential oral medroxyprogesterone acetate in postmenopausal women receiving continuous oral E2-valerate. DESIGN: One-year prospective multicentre randomised control trial. SETTING: Four outpatient clinics in Oulu, Helsinki and Jyvaskyla, Finland. POPULATION: A total of 163 healthy volunteer postmenopausal women with climacteric complaints or already using hormone replacement therapy (HRT). INTERVENTIONS: Subjects were randomly allocated to receive a new intrauterine system releasing 10 microg of levonorgestrel daily or an established intrauterine system (Mirena) releasing 20 microg of levonorgestrel daily or sequential oral medroxyprogesterone acetate (5mg/day, 14/30 days). All three regimens were combined with an oral daily dose of 2mg of E2-valerate. MAIN OUTCOME MEASURES: Bleeding patterns were assessed by diaries kept by the subjects. Endometrial effects were evaluated by histologic biopsies taken at the baseline and after six and 12 months of therapy. Serum concentrations of total, HDL and LDL cholesterol, triglycerides and lipoprotein(a) were determined at the baseline and after six and 12 months of therapy. RESULTS: Insertion of the smaller 10 microg levonorgestrel system was easy in 70% and difficult in 4% and that of Mirena was easy in 46% and difficult in 21% of the subjects. After six months of therapy, 43 (95.6%) of the 47 subjects receiving 10 microg levonorgestrel and 54 (98.2%) of the 55 subjects receiving 20 microg levonorgestrel had no bleeding, while the sequential medroxyprogesterone acetate regimen produced typical cyclic withdrawal bleedings. Endometrial hyperplasia was not observed in any of the treatment groups during the 12-month study. After 12 months of therapy, strong endometrial suppression was found in 46/47 and 55/55 of the subjects receiving 10 microg and 20 microg of levonorgestrel, respectively, while the endometrium was proliferative in 18/47 of the subjects in the medroxyprogesterone acetate group. Serum total cholesterol decreased in all treatment groups. HDL cholesterol increased in women receiving medroxyprogesterone acetate or the smaller intrauterine dose of levonorgestrel. CONCLUSIONS: Both intrauterine doses of levonorgestrel provided good endometrial protection in postmenopausal women on oestrogen replacement therapy. The advantage of the 10 microg system with a smaller size is the easier insertion of the system and a minimal attenuation of the favourable effects of oral oestrogen on the serum lipid profile. PMID- 11888096 TI - Can ultrasound replace ambulatory urodynamics when investigating women with irritative urinary symptoms? AB - OBJECTIVE: To determine whether transvaginal ultrasound measurement of bladder wall thickness could replace ambulatory urodynamics when investigating women with lower urinary tract dysfunction not explained by conventional laboratory urodynamic studies. DESIGN: A blinded prospective study. SETTING: Tertiary referral unit in a London teaching hospital. POPULATION: One hundred and twenty eight women referred for ambulatory urodynamics with equivocal laboratory urodynamic findings or whose symptoms were not explained by the laboratory urodynamic findings. METHODS: Transvaginal ultrasound assessment of bladder wall thickness was performed in three planes with an empty bladder prior to ambulatory urodynamics. Mean bladder wall thickness was calculated and the results analysed with respect to the ambulatory urodynamic diagnosis. MAIN OUTCOME METHODS: Mean bladder wall thickness in women with a normal ambulatory study or a diagnosis of detrusor instability, genuine stress incontinence (GSI) or mixed incontinence. RESULTS: Using a one way analysis of variance (ANOVA) bladder wall thickness was found to be significantly different in all diagnostic groups and this reached significance (P = 0.0001). There was no overlap in the 95% confidence intervals representing a diagnosis of detrusor instability or genuine stress incontinence. CONCLUSIONS: Transvaginal ultrasound assessment of mean bladder wall thickness is a sensitive screening tool, which can detect detrusor instability in those women with equivocal laboratory urodynamics. In women who have no evidence of GSI on laboratory studies, a cutoff of 6.0mm is highly suggestive of detrusor instability. However, in those women with GSI then ambulatory studies probably remain the investigation of choice. PMID- 11888097 TI - Knowledge, use and attitudes towards emergency contraceptive pills among Swedish women presenting for induced abortion. AB - OBJECTIVE: To investigate the knowledge, experiences and attitudes towards emergency contraceptive pills (ECP) among women presenting for induced abortion. DESIGN: Survey by self-administered waiting room questionnaires. SETTING: Three large hospitals in the cities of Uppsala, Vasteras and Orebro in Sweden. POPULATION: 591 Swedish-speaking women consecutively attending the clinics for an induced abortion during a four-month period in 2000. RESULTS: The response rate was 88% (n = 518). As many as 43% had a history of one or more previous abortions and 43% were daily smokers. Four out of five women, 83%, were aware of ECP, but only 15 women used it to prevent this pregnancy. Fewer, 38%, knew the recommended timeframes for use and 54% had knowledge of the mode of action. The two most common sources of information about ECP were media and friends. One out of five, 22%, had previously used the method, and at the time of conception, 55% would have taken ECP if it had been available at home, and 52% were positive to having ECP available over the counter. CONCLUSIONS: Emergency contraception is well known but is still underused. Lack of awareness of pregnancy risk may be one limiting factor for its use. Making ECP available over the counter may be an important measure towards better availability. Information strategies to the public are needed before ECP will be a widely used back-up method. PMID- 11888098 TI - Low dose acetylsalicylic acid in prevention of pregnancy-induced hypertension and intrauterine growth retardation in women with bilateral uterine artery notches. AB - OBJECTIVE: To evaluate the efficacy of low-dose acetylsalicylic acid in the prevention of pregnancy-induced hypertension and intrauterine growth retardation in high-risk pregnancies as determined by transvaginal Doppler ultrasound study of the uterine arteries at 12 to 14 weeks of gestation. DESIGN: Randomised, double blind and placebo-controlled trial. SETTING: The Department of Obstetrics and Gynaecology, Tampere University Hospital, Finland. POPULATION: One hundred and twenty pregnant women considered to be at high risk of pre-eclampsia or intrauterine growth retardation were screened by transvaginal Doppler ultrasound at 12 to 14 weeks of gestation. METHODS: Ninety pregnant women with bilateral notches in the uterine arteries were randomised to receive acetylsalicyclic acid 0.5mg/kg/day (n = 45) or placebo (n = 45) from 12 to 14 weeks of gestation. MAIN OUTCOME MEASURES: Hypertensive disorders of pregnancy and intrauterine growth retardation. RESULTS: Forty-three women on acetylsalicyclic acid and 43 on placebo were successfully followed up. The use of acetylsalicyclic acid was associated with a statistically significant reduction in the incidence of pregnancy-induced hypertension (11.6% vs 37.2%, RR = 0.31, 95% CI 0.13-0.78) and pre-eclampsia (4.7% vs 23.3%, RR = 0.2, 95% Cl 0.05-0.86). The incidence of hypertension before 37 weeks of pregnancy was also significantly reduced (2.3% vs 20.9%, RR = 0.22, 95% CI 0.05-0.97). The reduction in the incidence of intrauterine growth retardation (2.3% vs 7%) was not statistically significant. Acetylsalicyclic acid was not associated with excess risk of maternal or fetal bleeding. CONCLUSION: In women rated in Doppler velocimetry waveform analysis to be at high risk of pre-eclampsia, low-dose acetylsalicyclic acid reduces the incidence of pregnancy-induced hypertension and especially proteinuric pre eclampsia. PMID- 11888099 TI - A randomised comparison of early versus late amniotomy following cervical ripening with a Foley catheter. AB - OBJECTIVES: Ripening of the cervix with a Foley catheter commonly results in cervical dilatation without contractions. We examined the outcome of labour in women who underwent induction of labour using a Foley catheter, followed by either 1. early amniotomy, or 2. augmentation of labour by oxytocin and late amniotomy. DESIGN: Prospective randomised clinical trial. SETTING: Labour and delivery ward of a university teaching hospital. PARTICIPANTS: Pregnant women > or =38 weeks of a singleton gestation, who had had no prior caesarean section. METHODS: All women underwent cervical ripening using a Foley catheter. Following removal of the catheter, women were randomly assigned to either early (n = 80) or late amniotomy (n = 88). MAIN OUTCOME MEASURES: Comparison of mode of delivery and duration of labour between the two groups. RESULTS: The rate of caesarean section was significantly higher in the early amniotomy group compared with the late amniotomy group (25% vs 7.9%; relative risk 1.74; 95% CI 1.3 - 2.34). The increase in caesarean section rate was due primarily to dystocia (15% vs 3.3%; relative risk 1.8; 95% CI 1.32 - 2.45). When excluding caesarean deliveries, no significant difference was found in duration of labour between the groups (8.3 hours (3.8) vs 7.7 hours (2.9)). CONCLUSIONS: In women who undergo cervical ripening with a Foley catheter, augmentation of labour by oxytocin followed by amniotomy during active labour results in a lower rate of caesarean delivery for dystocia. PMID- 11888100 TI - Warfarin dosage in postpartum women: a case-control study. AB - OBJECTIVE: To investigate the clinical suspicion that postpartum women are more difficult to anticoagulate with warfarin than non-pregnant women due to the physiological changes in coagulation proteins that persist into the postpartum period. DESIGN: A retrospective case-control study. SETTING: University Hospital, Nottingham, UK. SAMPLE: Twenty-three postpartum women discharged from the obstetric wards on warfarin and 23 age-matched control women discharged from the medical wards on warfarin were identified using hospital databases. METHODS: Warfarin doses and international normalised ratio values were recorded from day one to 35. The number of days and total warfarin dose to achieve therapeutic international normalised ratio were recorded. Doses were compared with those recommended by a dosing nomogram. RESULTS: The postpartum group took significantly longer and significantly larger doses of warfarin to reach therapeutic international normalised ratio (P < 0.05). The postpartum group required a persistently higher maintenance dose of warfarin. Comparing the warfarin dose given on day three with a standardised nomogram, 79% of women in the postpartum group compared with 57% in the control group were under-dosed. CONCLUSION: Postpartum women require larger doses of warfarin to reach therapeutic international normalised ratio than non-pregnant women. We would recommend the use of a dosing nomogram. PMID- 11888101 TI - Is there an increased maternal-infant prevalence of Factor V Leiden in association with severe pre-eclampsia? AB - OBJECTIVE: To compare the prevalence of the Factor V Leiden mutation in children and maternal-infant pairs in pregnancies affected by severe pre-eclampsia with unmatched normal controls. DESIGN: Prospective cohort study. SETTING: Department of Women's and Children's Health, The Canberra Hospital, Garran, ACT, Australia. SAMPLE: Forty-eight maternal-infant pairs where the index pregnancy was affected by severe pre-eclampsia; 46 unmatched maternal-infant pairs where the index pregnancy was defined as normal. METHODS: DNA analysis of cheek swab samples obtained from maternal-infant pairs for the Factor V Leiden mutation. MAIN OUTCOME MEASURE: The prevalence of the Factor V Leiden mutation in mothers, infants and maternal-infant pairs in association with severe pre-eclampsia compared with unmatched controls. RESULTS: No difference was detected in the prevalence of Factor V Leiden mutation between the women and children of both groups, nor the maternal-infant pairs from each group. CONCLUSIONS: No evidence was found of an increased prevalence of the Factor V Leiden mutation in either the mothers or children in association with severe pre-eclampsia. This result argues against a Factor V Leiden fetal or maternal contribution to the development of severe pre-eclampsia. PMID- 11888102 TI - Adhesion molecules expression in the placental bed of pregnancies with pre eclampsia. AB - OBJECTIVE: To compare the expression of intercellular adhesion molecule-1 (ICAM 1), vascular cell adhesion molecule-1 (VCAM-1), platelet endothelial cell adhesion molecule-1 (PECAM-1) and E-selectin in placental bed biopsies (endothelium of spiral arteries as well as trophoblastic cells) from normal and pre-eclamptic pregnancies. DESIGN: A prospective study. SETTING: 1. First Department of Obstetrics and Gynaecology, Alexandra Hospital, University of Athens, Greece. 2. Laboratory of Pathophysiology, Laikon Hospital, University of Athens. POPULATION: Sixteen placental bed biopsies from women with pre-eclampsia were compared with 20 placental bed biopsies from uncomplicated normotensive women. MAIN OUTCOME MEASURES: Immunocytochemical staining of the placental bed biopsies for ICAM-1, VCAM-1, PECAM-1 and E-selectin. RESULTS: No statistically significant differences were found in the expression of the adhesion molecules ICAM-1, VCAM-1, PECAM-1 and E-selectin in the endothelium of the spiral arteries and the trophoblastic cells of the placental bed of the two studied groups. CONCLUSIONS: Adhesion molecules ICAM-1, VCAM-1, PECAM-1, but not E-selectin, are expressed in the placental bed of normal and pre-eclamptic pregnancies, but there are no differences between the two groups of women. It seems that the above molecules are not likely to be implicated in the aetiology of pre-eclampsia. PMID- 11888103 TI - Haemodynamic changes in the brain after vaginal delivery and caesarean section in healthy term infants. AB - OBJECTIVE: To investigate whether the mode of delivery may affect neonatal cerebral haemodynamics during the first hour of life. DESIGN: Prospective study. SAMPLE: Healthy infants with gestational age > or =37 weeks and birthweight appropriate for gestational age, born after uncomplicated pregnancy by vaginal delivery or elective caesarean section, two to five hours after the delivery. METHODS: Near infra-red spectroscopy was used to measure changes of oxygenated haemoglobin, deoxygenated haemoglobin, oxidized-reduced cytochrome aa3, and mean cerebral oxygen saturation (mixed cerebral oxygen saturation = oxygenated haemoglobin/total haemoglobin). Changes in cerebral blood volume were calculated. RESULTS: Near infra-red spectroscopy data did not show significant differences between infants born by vaginal delivery or by caesarean section. There was a significant decrease of oxygenated haemoglobin and change of cerebral blood volume values at 120 and 180 minutes in both the groups, while deoxygenated haemoglobin and oxidized-reduced cytochrome aa3 were unchanged. CONCLUSIONS: A decrease of cerebral blood volume occurs after birth and this occurs both in infants born by vaginal delivery and by caesarean section. PMID- 11888104 TI - The impact of an assisted conception unit on the workload of a general gynaecology unit. AB - The burden placed on a hospital by the presence of an assisted conception unit has been emphasised only in terms of its impact on neonatal services. This paper examines the previously neglected subject of the gynaecological workload generated by a tertiary fertility centre that provides treatments by assisted conception. As many IVF units operate independently this additional workload may not be appreciated. It has, however, significant practical and financial implications for neighbouring hospitals and trusts. This is of particular relevance in view of the move towards more uniform health service funding of assisted conception throughout the United Kingdom. PMID- 11888105 TI - The economic costs of alternative modes of delivery during the first two months postpartum: results from a Scottish observational study. AB - A study was conducted to estimate the economic costs of alternative modes of delivery during the first two months postpartum. Hospital and community health service utilisation data for 1242 women were extracted from self-completed questionnaires, medical case notes and computerised hospital discharge records. Unit costs (1999-2000 prices) were collected for each item of resource use and combined with resource volumes to obtain a net cost per woman. There were significant differences in initial hospitalisation costs between the three mode of delivery groups (spontaneous vaginal delivery pounds sterling 1431, instrumental vaginal delivery pounds sterling 1970, caesarean section pounds sterling 2924, P < 0.001). There were also significant differences in the cost of hospital readmissions, community midwifery care and general practitioner care between the three mode of delivery groups. However, total post-discharge health care costs did not vary significantly by mode of delivery. Total health care costs were estimated at pounds sterling 1698 for a spontaneous vaginal delivery, pounds sterling 2262 for an instrumental vaginal delivery and pounds sterling 3200 for a caesarean section (P < 0.001). It is imperative that hospital and community health service providers recognise the economic impact of alternative modes of delivery in their service planning. PMID- 11888106 TI - Rescue of trophoblast apoptosis by parathyroid hormone-related protein. AB - The effects of the 1-34 N-terminal parathyroid hormone-related protein fragment (0-10 microM) and the 67-86 parathyroid hormone-related protein fragment (0-10 microM) on trophoblast apoptosis, induced by TNFalpha and IFN-gamma or by the protein kinase inhibitor staurosporine, were investigated. TNFalpha/IFN-gamma and staurosporine significantly increased the rate of apoptosis by fivefold and by eightfold, respectively. Parathyroid hormone-related protein (1-34) evoked a dose dependent rescue of both TNFalpha/IFNgamma-induced and staurosporine-induced apoptosis, whereas parathyroid hormone-related protein (67-86) had no significant effect on staurosporine-induced apoptosis, and only significantly diminished TNFalpha/IFNgamma-induced apoptosis at 10 microM. Parathyroid hormone-related protein was thus found to be a cytotrophoblast apoptosis survival factor. PMID- 11888107 TI - May-hegglin anomaly: the role of aspirin in the treatment of this rare platelet disorder in pregnancy. PMID- 11888108 TI - Convictions and alleged misconduct lead to disciplinary inquiry. PMID- 11888109 TI - Support needs of veterinary surgeons during the first few years of practice: perceptions of recent graduates and senior partners. AB - Postal surveys or personal interviews of 76 recent veterinary graduates and their 49 employers were undertaken to establish their perceptions of good practice when integrating a new graduate into a business and their preferred methods of assessment and development. Practice type and location were the main influences on graduates looking for their first job. Interviews were mostly informal. Employers expected basic veterinary competence and candidates expected good quality support. Most graduates (93 per cent) had their own consultations on the first day. During early consultations 2 per cent of senior vets accompanied the new graduate, 95 per cent of practices provided senior back-up either in person or by telephone but in 3 per cent no back-up was available. Most new graduates (90 per cent) were satisfied with their workload. Three-fifths were on-call within the first week, and 95 per cent within a month. Graduates received calls directly in 45 per cent of practices, in 9 per cent seniors screened the calls, and the remainder used a third party. Assistance from experienced lay staff varied greatly. Discussion of problems was mainly informal. There was little spontaneous feedback and problems resulted from inadequate communication. One in three new graduates left their first job within two years, and one in six identified lack of support, heavy workload, stress or clashes with staff as a primary reason. This high turnover was a problem for employers. From the new graduates' perspectives, initial problems included: being on call (59 per cent), financial aspects (47 per cent) and surgery (43 per cent). Communicating with clients and learning to prioritise jobs were also difficult. New graduates took longer over procedures (79 per cent of employers commented) and required extra back-up (91 per cent) both of which reduced income (59 per cent). Nearly all the seniors felt that their current new graduates had coped 'quite well', although it was claimed that new graduates lacked the ability to talk to clients at the appropriate level, wanted to bring all their scientific knowledge to bear on every case, and often failed to consider the obvious or to appreciate clients' needs. Only 18 per cent of practices had formal and regular review procedures but all monitored the response of clients and watched the new graduate perform. Feedback to their new colleague was considered 'adequate' by 85 per cent of seniors, although 45 per cent of graduates felt they had not received enough. Eighty-three per cent of new graduates felt 'moderately prepared' by their undergraduate course, and 76 per cent of senior vets were 'generally satisfied' Both wanted improvements in extramural studies and increased exposure to routine cases. Senior partners sought greater commitment in the undergraduate curriculum to financial/legal issues and communication skills. Over a third of employers (38 per cent) had a 'great influence' on the choice of continuing professional development courses for their recent graduates. New graduates chose courses to deal with a perceived weakness, or to specialise, and welcomed opportunities to meet other new graduates and share early experiences. It was concluded that turnover and staff problems would be reduced if practices became more effective in coping with new arrivals, especially by supporting their development. PMID- 11888110 TI - Effect of electrocardiographic filters on the R-amplitude of canine electrocardiograms. AB - Lead II electrocardiograms (ECGs) were recorded in 88 dogs ranging in weight from 3 to 50 kg. A commercial direct-writing electrocardiograph was used to record the ECGs, first with manual HUM filter (notch filter at 50/60 Hz) and EMG filter (cut off frequency of -3 dB at 35 Hz) on, immediately followed by a recording with both filters off. The mean (sd) reduction in R-amplitude with the filters on was 53(18) per cent with a range from 22 to 100 per cent and a median value of 51 per cent. The R-amplitude with the filters off was related to the R-amplitude with the filters on, making it possible to correct for the effect of the filters. The reduction in R-amplitude was inversely correlated with the weight of the dog and with the duration of the QRS-complex. Other known changes induced by filters, such as the elimination of notches and the slurring of the junction between the QRS-complex and the ST-interval were also observed. The inverse relationship between the weight of the dog and the magnitude of the reduction in R-amplitude by the filters indicates that ECG machines with different characteristics should ideally be used to record ECGs in dogs of different sizes. PMID- 11888111 TI - Detecting antibodies to infectious bovine rhinotracheitis and BVD virus infections using milk samples from individual cows. PMID- 11888112 TI - Efficacy of pyrantel embonate, febantel and praziquantel against Giardia species in naturally infected adult dogs. PMID- 11888113 TI - Isolation of Brucella melitensis from an arabian oryx (Oryx leucoryx). PMID- 11888114 TI - Duty of care regarding night visits. PMID- 11888115 TI - Duty of care regarding night visits. PMID- 11888116 TI - Veterinary nurse training. PMID- 11888117 TI - Veterinary nurse training. PMID- 11888118 TI - Parvovirus in pigs with multifocal interstitial nephritis. PMID- 11888119 TI - Gasterophilus in dogs. PMID- 11888120 TI - Access to the guide to professional conduct. PMID- 11888121 TI - Nursing, our public deaths, and the tobacco industry. PMID- 11888122 TI - "Nurse, I only had a couple of beers": validity of self-reported drinking before serious vehicular injury. AB - BACKGROUND: Of the more than 40000 Americans killed each year in vehicular crashes, 40% are involved in alcohol-related collisions. Although self-reported alcohol use has become an anchor for alcohol intervention after traffic crashes, clinicians are often skeptical about the truthfulness of self-reporting. OBJECTIVE: To determine the validity of self-reported alcohol consumption of vehicular occupants hospitalized for a serious, alcohol-related injury. METHODS: Non-alcohol-dependent subjects 18 years and older who were injured in motor vehicle crashes were interviewed. The self-reported number of standard drinks, time that drinking commenced, sex, and weight were used to calculate estimated blood alcohol concentration. This value was compared with the blood alcohol concentration measured at admission. RESULTS: Of the 181 subjects, 60% provided sufficient data to calculate the estimated concentration. Seven men with admission concentrations of 10 mg/dL or more denied drinking. Among the 113 subjects with estimated concentrations who acknowledged drinking (excluding the 7 who denied drinking), the mean concentration at admission was 158.67 mg/dL, and mean estimated concentration was 83.81 mg/dL. According to multiple regression analyses, weight and number of drinks accounted for 3% of the variance in alcohol concentration at admission for women (R=0.174, F2,40=0.623, P=.54) and for 29% of the variance in men (R=0.543, F2,128=26.71, P< .001). CONCLUSIONS: Most persons who drink before vehicular injury acknowledge drinking. Self-reported data from men generally reflect the overall trend of alcohol consumption but with systematic underreporting. Reports from women are less predictable. PMID- 11888123 TI - Hospital experiences of young adults with congenital heart disease: divergence in expectations and dissonance in care. AB - BACKGROUND: Despite increasing survival for adults with congenital heart disease, little is known about hospitalization for young adult patients with this disease and for their families. Because of the complexity of the disease and its management during the life span, young adults are often hospitalized on both pediatric and adult units during a stay in the hospital. OBJECTIVES: To explore the experience of hospitalization of young adults with congenital heart disease, the experience of their families, and the views of the nurses who cared for these patients and to generate substantive theory on interactions between patients, patients' families, and nurses. METHODS: Semistructured interviews and naturalistic observations were conducted with young adults with congenital heart disease (mean age, 28.6 years), their family members, and nurses who cared for the patients during hospitalization (N=34). Dimensional analysis was used to analyze interviews and field notes from observations. RESULTS: A grounded theory was derived, explaining how the hospital context and relationships between patients, patients' families, and nurses affect patients' hospital experiences. Expectations differed among the groups, leading to dissonance in care, as exemplified by role confusion and power struggles over control of care. This dissonance resulted in interpersonal conflict, distrust, anxiety, and dissatisfaction with the care and caring experiences. CONCLUSIONS: Changes in hospital units, a better understanding of the healthcare needs of young adults with congenital heart disease, and acknowledgment of the expertise of patients and patients' families are needed to improve nursing care for these patients and their families. PMID- 11888124 TI - Accidental carbon monoxide poisoning with severe cardiorespiratory compromise in 2 children. PMID- 11888125 TI - Using a collaborative weaning plan to decrease duration of mechanical ventilation and length of stay in the intensive care unit for patients receiving long-term ventilation. AB - BACKGROUND: Patients requiring mechanical ventilation for prolonged periods typically are sicker and have more comorbid illnesses than do patients who can be weaned more rapidly. As a result, the weaning process is often complex, requiring shared decision making by a skilled, multidisciplinary team. Unfortunately, many of the structures used in critical care units to plan and evaluate care do not lend themselves to collaborative management of patients. OBJECTIVE: To evaluate the effect of a collaborative weaning plan on outcomes, including duration of mechanical ventilation, for patients treated with mechanical ventilation for 7 days or more. METHODS: A collaborative weaning plan (weaning board and flow sheet) was introduced into the medical intensive care unit at the University of California Los Angeles, Medical Center. A historical design was used to compare outcomes before and after the plan was used. The primary outcome variable was duration of mechanical ventilation. Other outcomes studied included length of stay in the unit, cost, prevalence of complications (ie, reventilation, readmission to the intensive care unit), and mortality rate. RESULTS: The collaborative weaning plan decreased duration of ventilation by 4.9 days (P=.02) and decreased median length of stay in the unit by 4.5 days (P=.004). The median cost per stay in the unit decreased from $50462 to $37330 (P=.004). The prevalence of complications did not differ significantly between groups. CONCLUSIONS: Collaborative structures (eg, weaning boards, flow sheets) are useful in decreasing duration of mechanical ventilation for patients receiving long-term ventilation. PMID- 11888126 TI - Bacterial growth in secretions and on suctioning equipment of orally intubated patients: a pilot study. AB - BACKGROUND: Contamination of equipment, colonization of the oropharynx, and microaspiration of secretions are causative factors for ventilator-associated pneumonia. Suctioning and airway management practices may influence the development of ventilator-associated pneumonia. OBJECTIVES: To identify pathogens associated with ventilator-associated pneumonia in oral and endotracheal aspirates and to evaluate bacterial growth on oral and endotracheal suctioning equipment. METHODS: Specimens were collected from 20 subjects who were orally intubated for at least 24 hours and required mechanical ventilation. At baseline, oral and sputum specimens were obtained for culturing, and suctioning equipment was changed. Specimens from the mouth, sputum, and equipment for culturing were obtained at 24 hours (n=18) and 48 hours (n=10). RESULTS: After 24 hours, all subjects had potential pathogens in the mouth, and 67% had sputum cultures positive for pathogens. Suctioning devices were colonized with many of the same pathogens that were present in the mouth. Nearly all (94%) of tonsil suction devices were colonized within 24 hours. Most potential pathogens were gram positive bacteria. Gram-negative bacteria and antibiotic-resistant organisms were also present in several samples. CANCLUSIONS: The presence of pathogens in oral and sputum specimens in most patients supports the notion that microaspiration of secretions occurs. Colonization is a risk factor for ventilator-associated pneumonia. The equipment used for oral and endotracheal suctioning becomes colonized with potential pathogens within 24 hours. It is not known if reusable oral suction equipment contributes to colonization; however, because many bacteria are exogenous to patients' normal flora, equipment may be a source of cross-contamination. PMID- 11888127 TI - Pepsin as a marker for pulmonary aspiration. AB - BACKGROUND: Although assessment for aspiration of small volumes of gastric contents in tube-fed patients receiving mechanical ventilation is important, available methods for this purpose are not wholly satisfactory. A potential method is immunoassay of tracheal secretions for the gastric enzyme pepsin. OBJECTIVES: To determine the frequency with which pepsin in suctioned tracheal secretions from acutely ill, tube-fed patients receiving mechanical ventilation could be detected via an immunoassay. METHODS: A convenience sample of 136 specimens of suctioned tracheal secretions was collected from 30 acutely ill, tube-fed adults receiving mechanical ventilation. Multiple samples were obtained from 26 of the 30 patients (range, 2-11 per subject). An immunoassay with rooster polyclonal antibodies to purified human pepsin was used to detect pepsin in the secretions. RESULTS: Fourteen specimens tested positive for pepsin. Secretions from 5 patients accounted for the 14 pepsin-positive results. A significant relationship was found between the position of the head of the bed and the presence of pepsin in tracheal secretions (P<.001). Of the 14 pepsin-positive specimens, 13 (92.9%) were obtained from subjects in a flat position. CONCLUSIONS: A pepsin immunoassay can be used to detect pepsin in human tracheal secretions. If pepsin in tracheal secretions is considered an indicator of aspiration of gastric contents, aspiration occurred in 5 of the 30 subjects. A flat position is strongly associated with the presence of pepsin in tracheal secretions. PMID- 11888129 TI - Creating compassionate institutions that foster agency and respect. PMID- 11888128 TI - Effectiveness of mechanical compression devices in attaining hemostasis after femoral sheath removal. AB - BACKGROUND: Cardiac interventions are widely accepted as a practical treatment option for coronary artery disease. However, few changes have occurred in the techniques used for percutaneous arterial cannulation and for attaining hemostasis after cardiac interventions. To date, researchers have focused on techniques to achieve optimal hemostasis at the time of removal of the arterial catheter and to minimize the impact and complications of arterial puncture. OBJECTIVE: To summarize the best available evidence on the effectiveness of mechanical compression devices used to obtain hemostasis following femoral sheath removal after cardiac interventional procedures. METHOD: An attempt was made to detect both published and unpublished reports of research evaluations of mechanical compression techniques used to attain hemostasis after femoral sheath removal. Methodological quality was assessed by using predesigned criteria. Data were extracted from information on randomized controlled trials and were statistically combined in meta-analysis where possible. Evidence was also synthesized by using narrative summaries. RESULTS: Twelve studies met the inclusion criteria; however, only 3 were included in the meta-analysis. The results of meta-analysis indicated that the mechanical compression technique was the most effective for preventing formation of hematomas. The prevalence of bleeding did not differ significantly for different methods of compression. CONCLUSION: A gap exists in the literature on quality randomized controlled trials of various devices used to attain hemostasis after femoral sheath removal. PMID- 11888130 TI - Novel markers in the acute coronary syndrome: BNP, IL-6, PAPP-A. AB - Most patients (about 85%) seen in the ED to rule out an acute coronary event do not have acute coronary disease. In addition, the presenting ECG findings have been nondiagnostic in 50% of patients with acute MI. Our current knowledge of atherosclerosis as being a chronic low-grade inflammatory process triggered the search for reliable serum markers that have improved the diagnostic accuracy management and prognosis of this prevalent disease. Newer and potential inflammatory markers currently under investigation deserve watching in future reports. These among others include those markers produced by the arterial wall itself, that is, cell adhesion molecules (CAM), inter-cellular adhesion molecules (ICAM), and vascular adhesion molecules (VCAM). The expression of CAM is a marker of dysfunctional endothelial cells. It is likely that more cardiac markers will be reported in the future. Time will tell. PMID- 11888131 TI - Inverted P waves. PMID- 11888132 TI - On notebooks and trust. PMID- 11888133 TI - Anatomy, pathophysiology, and biomechanics of shoulder instability. AB - Instability in the athlete presents a unique challenge to the orthopaedic surgeon. A spectrum of both static and dynamic pathophysiology, as well as gross and microscopic histopathology, contribute to this complex clinical continuum. Biomechanical studies of the shoulder and ligament cutting studies in recent years have generated a more precise understanding of the individual contributions of the various ligaments and capsular regions to shoulder instability. An understanding of the underlying pathology and accurate assessment of degree and direction of the instability by clinical examination and history are essential to developing appropriate treatment algorithms. PMID- 11888134 TI - Differential diagnosis of shoulder injuries in sports. AB - Shoulder injuries are common problems in all types of sports. There can be a good deal of variability in how symptoms are expressed from individual to individual, and part of the challenge in understanding the injury is being able to interpret the athlete's complaints as they relate to examination findings and the conditions that produce the symptoms. The history is particularly important because the sensitivity and specificity of many aspects of the examination are variable. A relaxed patient and a systematic evaluation is the best way to be thorough. Radiographs and even MRIs are frequently negative in soft tissue problems in the young athlete. This underscores the importance of a good examination. There may be some utility in using contrast-enhanced MRIs to look at labral pathologic conditions, cysts, or ganglions, but this information is certainly less useful than the patient's description of symptoms and the examiner's findings. With this awareness, we can manage these injuries with more confidence. PMID- 11888135 TI - Traumatic anterior shoulder instability. PMID- 11888136 TI - Arthroscopic Bankart repair with the Suretac device for traumatic anterior shoulder instability in athletes. AB - Arthroscopic treatment of anterior shoulder instability in the athlete has evolved tremendously over the past decade. Currently, most techniques include the use of suture and suture anchors. However, the variety of arthroscopic instruments and techniques that are available shows the complexity of intra articular tissue fixation, which includes anchor placement, suture passing, and knot tying. Stabilization using the Suretac device (Acufex Microsurgical, Mansfield, MA) simplifies tissue fixation by eliminating the need for arthroscopic suture passing and intra-articular knot tying. However, a successful outcome is highly dependent on accurate patient selection. Preoperative evaluation, examination under anesthesia, and the pathoanatomy defined by a thorough arthroscopic examination suggest the most effective treatment strategy. The ideal candidate for shoulder stabilization using the Suretac device is an athlete with a relatively pure traumatic anterior instability pattern with detachment pathology (e.g., Bankart lesion) and minimal capsular deformation. PMID- 11888137 TI - Evaluation and management of shoulder instability in the elite overhead thrower. AB - The elite throwing athlete places significant forces on the soft-tissue stabilizers of the shoulder with every pitch. Anterior translation forces can be as high as 40% of body weight and distraction forces as high as 80% body weight during the act of throwing. Injury to the static and dynamic stabilizers can lead to significant pain and loss of function in these athletes. To successfully treat the injured thrower, it is important to accurately diagnose the pathologic process. This article reviews the biomechanics of throwing and pathologic processes seen in the elite thrower. We cover the essentials of the history and physical in this population and conclude with a discussion of the various treatment regimens. PMID- 11888138 TI - SLAP lesions in the overhead athlete. AB - The authors report an 87% rate of return to preinjury levels of throwing in 54 baseball players and an 84% rate of return to preinjury performance levels in pitches after repair of type II SLAP lesions. The etiology, biomechanics, surgical repair, and rehabilitation are discussed in detail. PMID- 11888139 TI - Arthroscopic repair of partial-thickness rotator cuff tears and SLAP lesions in professional baseball players. AB - Internal impingement is a primary cause of shoulder pain in throwers; however, instability, internal rotation deficit, scapula muscle dysfunction, and core muscle dysfunction are also important elements of the internal impingement process. Articular surface rotator cuff tears, posterior superior labrum tears, SLAP lesions, anterior capsular ligament attenuation, and posterior capsular ligament contracture are commonly seen in throwers. Each of these conditions must be recognized and appropriately treated to ensure the best possible outcome. There is little potential for spontaneous healing of rotator cuff tears and SLAP lesions after debridement. PMID- 11888140 TI - Anterior superior instability with rotator cuff tearing: SLAC lesion. AB - Anterosuperior instability of the shoulder may occur from a variety of pathologic lesions. We describe a specific entity, the SLAC (superior labrum, anterior cuff) lesion that involves an association of anterior-superior labral tear with a partial supraspinatus tear. We retrospectively isolated a group of 40 patients with this lesion. The presenting complaints, physical examination findings, surgical findings, and results were isolated. Overhead activities were the most common etiology; load and shift instability testing and whipple rotator cuff testing were the most common physical examination findings. Surgical repair was successful in 37 of the 40 patients. The SLAC lesion is a definable clinical entity with predictable history, examination, surgical pathology, and satisfactory results from surgery. PMID- 11888141 TI - Posterior instability. AB - Posterior shoulder instability is a pathology that is increasingly seen in athletes. Excessive capsular laxity was originally proposed as the key component. Recent cadaveric and arthroscopic work has identified the importance of glenolabral integrity and glenoid depth in maintaining glenohumeral stability. Arthroscopic techniques to treat posterior instability are emerging. Until recently, reports of arthroscopic reconstruction focused entirely on capsular glenohumeral stability by altering two separate mechanisms: deepening of the glenoid concavity and reducing the capsular joint volume. This is accomplished by shifting the capsule to buttress the glenoid labrum. Thus increasing capsular tension increases the resultant compressive force vector into a deepened glenolabral concavity that, when combined together, enhances glenohumeral stability. In clinical and laboratory settings, we have shown that posteroinferior shoulder instability is associated with both capsular laxity and well-defined pathological lesions of the glenolabral concavity. Our results indicate that arthroscopic posterior capsulolabral repair and augmentation is a useful tool to restore the depth of the glenolabral concavity and to reduce the redundant posteroinferior capsule. This technique is effective in treating posteroinferior instability. PMID- 11888143 TI - Coracoid impingement syndrome, rotator interval reconstruction, and biceps tenodesis in the overhead athlete. AB - Anterior shoulder problems are extremely common in throwing athletes. Coracoid impingement syndrome, lesions of the long head of the biceps tendon, and rotator interval lesions are included in the extensive differential diagnosis which exists for anterior shoulder pain. In this article, we focus on the anatomy, pathophysiology, clinical presentation, diagnosis, and surgical treatment of these conditions. PMID- 11888142 TI - Treatment of the athlete with multidirectional shoulder instability. AB - Treatment of multidirectional instability in the athlete continues to be a challenging problem. Nonoperative treatment remains the initial treatment of choice in most athletes. However, some patients will fail nonoperative treatment and might require operative intervention. An open capsular shift procedure is indicated in nonoverhead athletes, most contact athletes and in most revision situations. However, the advances in thermal treatment of the capsule have made arthroscopic electrothermal capsulorrhaphy increasingly attractive as an alternative to the open approach for the primary treatment of multidirectional instability. Longer follow-ups will be necessary before definitive statements can be made regarding the arthroscopic techniques. PMID- 11888144 TI - Technique for repair of the subscapularis tendon. AB - Patients with sudden loss of active motion after an external rotation or hyperextension injury should be viewed with a high index of suspicion for a subscapularis tear. Exaggerated external rotation and the presence of a positive lift off or belly press test on physical examination combined with appropriate imaging studies will lead to an early diagnosis. Careful surgical repair combined with a thoughtful rehabilitation program will lessen both the length and degree of disability from this clinical entity. PMID- 11888145 TI - All arthroscopic rotator cuff repairs. AB - The arthroscopic operation for repair of full-thickness rotator cuff tears is successful and has the advantages of glenohumeral joint inspection, treatment of intra-articular lesions, smaller incisions, no deltoid detachment, less soft tissue dissection, less pain, and more rapid rehabilitation. However, these advantages must be balanced against the technical difficulty of this method, which limits its application to surgeons skilled in both open and arthroscopic shoulder operations. This article contains many technical pearls to, as much as possible, simplify and improve all arthroscopic cuff repair. PMID- 11888146 TI - Monopolar radiofrequency energy for arthroscopic treatment of shoulder instability in the athlete. AB - Monopolar radiofrequency energy is increasingly being used in the treatment of shoulder instability. Basic science studies and early clinical results have shown that application of thermal energy can result in successful shrinkage of the shoulder capsule. This procedure is useful in treating certain traumatic and recurrent instability conditions of the shoulder especially in the athlete where range of motion is preserved, recovery is faster than with open procedures, and there, is little disruption or alteration of inherent anatomy. The procedure is technically easy to perform, and the complication rate is low. Success, however, depends on proper patient selection, attention to the rehabilitation program, and patient compliance. Long-term follow-up will be necessary to determine if results for this procedure will deteriorate over time, especially in patients with multidirectional instability. PMID- 11888147 TI - Technical note: a "new" arthroscopic sliding knot. AB - Sliding knots are an essential element of arthroscopic shoulder surgery. The authors have been using a previously undescribed arthroscopic sliding knot with good clinical success. This knot has been used in arthroscopic rotator cuff repair, arthroscopic shoulder stabilization, and arthroscopic SLAP repair. The technique of this knot is illustrated. PMID- 11888148 TI - Shoulder rehabilitation strategies, guidelines, and practice. AB - This framework for rehabilitation is consistent with the proximal-to-distal kinetic chain biomechanical model and applies current concepts of motor control and closed chain exercises. This framework approaches the final goal- glenohumeral motion and function-through facilitation by scapular control, and scapular control through facilitation by hip and trunk activation. This article supplies guidelines for rehabilitation and practices to implement the guidelines that have proved effective in our hands. Other protocols may be effective, as long as they adhere to several basic concepts of kinetic chain-based shoulder rehabilitation: 1. Functional shoulder rehabilitation requires that the muscle activations and joint motions follow a proximal-to-distal pathway along the appropriate kinetic chain. 2. Muscles around the shoulder function in an integrated fashion and should be rehabilitated in integrated patterns. Specific muscles may need isolated activation, but this activation should be facilitated by placing the proximal segments in a facilitating function. 3. Scapular control and coupled rotator cuff activation is vital to normal shoulder function. 4. Closed chain axial loading exercises are the primary means of early shoulder rehabilitation and are the mainstays of functional rehabilitation protocols. PMID- 11888149 TI - Tailoring tolerance to prevent chronic rejection. PMID- 11888150 TI - Monitoring the patient off immunosuppression. Conceptual framework for a proposed tolerance assay study in liver transplant recipients. AB - The mission of the recently established Immune Tolerance Network includes the development of protocols for the induction of transplant tolerance in organ allograft recipients and the development of assays that correlate with and may be predictive of the tolerant state. The state of clinical organ transplant tolerance seems to already exist in a small minority of conventionally immunosuppressed liver and, more rarely, kidney transplant patients. Immunosuppressive drug therapy has been withdrawn from these patients for a variety of reasons, including protocolized weaning for a uniquely large group of liver patients at the University of Pittsburgh. In this study, we propose to evaluate the validity of a variety of in vitro immunologic and molecular biologic tests that may correlate with, and be predictive of, the state of organ transplant tolerance in stable liver patients off immunosuppression. Only peripheral blood will be available for the execution of these tests. Both adult and pediatric liver graft recipients will be studied, in comparison to appropriate controls. We shall examine circulating dendritic cell (DC) subsets [precursor (p) DC1 and p DC2] including cells of donor origin, and assess both the frequency and function of donor-reactive T cells by ELISPOT and by trans-vivo delayed-type hypersensitivity analysis in a surrogate murine model. Cytokine gene polymorphism and alloantibody titers will also be investigated. It is anticipated that the results obtained may provide physicians with a tolerance assay "profile" that may determine those patients from whom immunosuppressive therapy may be safely withdrawn. PMID- 11888151 TI - Tolerance and the microenvironment. PMID- 11888152 TI - The thymus and central tolerance. PMID- 11888153 TI - Infectious tolerance? Mechanisms and implications. PMID- 11888154 TI - Minimization of immunosuppression in kidney transplantation. The need for immune monitoring. PMID- 11888155 TI - Mixed allogeneic chimerism. Past, present, and prospects for the future. PMID- 11888156 TI - Islet cell transplantation tolerance. AB - Curative islet transplantation for type 1 diabetes currently requires lifelong systemic immunosuppression. Induction of islet transplantation tolerance would be far preferable. We have previously demonstrated that blockade of costimulation by the administration of a donor-specific transfusion in combination with anti-CD154 monoclonal antibody leads to permanent islet and prolonged skin allograft survival in mice. The protocol requires the presence of CD4+ T cells, interferon gamma, and CTLA4, and involves the deletion of CD8+ alloreactive T cells. Translation of this strategy into clinical practice will, however, require attention to at least two issues. First, we have observed that the presence of viral infection during tolerance interferes with tolerance induction. Second, we have observed that our tolerance induction protocol is ineffective in autoimmune nonobese diabetic mice. We hypothesize that resistance to tolerance induction in nonobese diabetic mice is due to the presence of memory autoreactive cells. To overcome the deleterious effects of viral infection and of primed memory responses, it may be necessary to modify current tolerance induction strategies based on costimulatory blockade. These modifications may require patient isolation, the generation of hematopoietic chimerism, or treatments that target the specific T-cell populations, cytokines, and/or costimulatory factors responsible for resistance. Such modifications may make it possible to extend tolerance induction to the "real world" situation of individuals with type 1 diabetes who are likely to harbor both memory allo-and autoreactive immune cells. PMID- 11888157 TI - Immune self-tolerance mechanisms. AB - Tolerance of both T and B cells to self-antigens can be achieved through a great variety of different routes, at the level of the primary lymphoid organs (thymus and bone marrow) and throughout the secondary lymphoid tissues. Whether self reactive lymphocytes ignore their target autoantigen, or are tolerized by the various mechanisms discussed, depends on the circumstances. PMID- 11888158 TI - Simplified media for spiroplasmas associated with tabanid flies. AB - Traditionally, isolation, maintenance, and testing of Spiroplasma species (Mollicutes: Entomoplasmatales) from horse flies (Tabanus spp.) and deer flies (Chrysops spp.) (Diptera: Tabanidae) have been accomplished in the complex M1D medium. A relatively inexpensive, simplified medium for tabanid spiroplasmas could expedite procedures that require large quantities of growth medium. Nine strains of spiroplasmas, eight from tabanids and one from mosquitoes, were cultured in three simplified broth media, R2, R8-1, and C-3G, and in M1D. There was no significant difference in the rate of spiroplasma growth in M1D and the three simplified media. R2 medium supported the growth of tabanid spiroplasmas more consistently and with better morphology through 10 subcultures than did the other simplified media. Primary isolations were made in R2 medium from tabanids collected (i) in Georgia, U.S.A., with 10 isolations from 10 flies and (ii) in coastal Costa Rica, with isolation rates of 70% (28/40) and 73% (27/37), respectively, for R2 and M1D. Of the seven group VIII field isolates from Costa Rica, four were capable of sustained growth in R2, and three were triply cloned in this simplified medium. These results suggest that the simplified medium R2 is suitable for many procedures with tabanid spiroplasmas. PMID- 11888159 TI - Effect of a fibrolytic enzyme preparation from Trichoderma longibrachiatum on the rumen microbial population of dairy cows. AB - The effects of supplementing a dairy cow diet with incremental levels of a fibrolytic enzyme preparation (preparation B) from Trichoderma longibrachiatum on the rumen microbial population were investigated. Two cows fitted with rumen cannulae were each fed a diet containing barley-based concentrate (52%), maize silage (29%), and chopped alfalfa hay (19%), supplemented with 0, 1, 2, 5, or 10 L of preparation B per tonne of dry matter (DM). Preparation B stimulated numbers of total viable bacteria in a quadratic manner (P < 0.05), to approximately 230, 330, 390, and 250% at 1, 2, 5, and 10 L x t(-1) DM, respectively. Preparation B increased the numbers of cellobiose-utilizing (P < 0.01), xylanolytic (P < 0.05), and amylolytic bacteria (P < 0.05), but had no effect (P > 0.05) on numbers of cellulolytic bacteria. However, when bacterial numbers enumerated on each substrate were expressed as a proportion of total viable bacterial numbers, only cellobiose utilizers were stimulated, and this stimulation was limited to the 1 L x t(-1) DM level of preparation B (P < 0.05). The results of this study demonstrate that the inclusion of an exogenous fibrolytic enzyme preparation in dairy cow diets increased the numbers of rumen bacteria that utilize hemicelluloses and secondary products of cellulose digestion. PMID- 11888160 TI - Methanogenic potential of tailings samples from oil sands extraction plants. AB - Approximately 20% of Canada's oil supply now comes from the extraction of bitumen from the oil sands deposits in northeastern Alberta. The oil sands are strip mined, and the bitumen is typically separated from sand and clays by an alkaline hot water extraction process. The rapidly expanding oil sands industry has millions of cubic metres of tailings for disposal and large areas of land to reclaim. There are estimates that the consolidation of the mature fine tails (MFT) in the settling ponds will take about 150 years. Some of the settling ponds are now evolving microbially produced methane, a greenhouse gas. To hasten consolidation, gypsum (CaSO4 x 2H2O) is added to MFT, yielding materials called consolidated or composite tailings (CT). Sulfate from the gypsum has the potential to stimulate sulfate-reducing bacteria (SRB) to out-compete methanogens, thereby stopping methanogenesis. This investigation examined three MFT and four CT samples from three oil sands extractions companies. Each was found to contain methanogens and SRB. Serum bottle microcosm studies showed sulfate in the CT samples stopped methane production. However, if the microcosms were amended with readily utilizable electron donors, the sulfate was consumed, and when it reached approximately 20 mg/L, methane production began. Some unamended microcosms were incubated for 372 days, with no methane production detected. This work showed that each MFT and CT sample has the potential to become methanogenic, but in the absence of exogenous electron donors, the added sulfate can inhibit methanogenesis for a long time. PMID- 11888161 TI - Examination of adhesive determinants in three species of Lactobacillus isolated from chicken. AB - The microbial adhesion process includes passive forces; electrostatic interactions; hydrophobic, steric forces; lipoteichoic acids; and specific structures, such as external appendages (lectins) and (or) extracellular polymers. In a previous work, we showed that Lactobacillus animalis, L. fermentum, and L. fermentum ssp. cellobiosus had lectinlike proteic structures on their surfaces and high hydrophobicity values on the cell surface of L. fermentum ssp. cellobiosus. Here, we examined the presence of the bacterial forces or structures that could be involved in the interaction between bacteria and epithelial cells. Lactobacillus animalis and L. fermentum possessed a net negative surface charge, whereas L. fermentum ssp. cellobiosus showed similar affinity to both cationic and anionic exchange resins, aggregated in the presence of ammonium sulfate, and had high affinity (75.4%) to a hydrophobic matrix. Only L. animalis was shown to have ribitol teichoic acids in the cell wall. The amount of polysaccharides from cell walls varied between different strains, with L. fermentum ssp. cellobiosus having the highest concentration. Lectin extracts obtained from lactobacilli did not possess sugar residues, thereby demonstrating the proteic nature of the superficial surface structures of three strains. The lactic acid bacteria studied here showed different surface determinants, which could be involved in the interactions between these lactobacilli and intestinal epithelial cells. PMID- 11888162 TI - High-level heterologous expression and secretion in Streptomyces lividans of two major antigenic proteins from Mycobacterium tuberculosis. AB - Two major antigens from Mycobacterium tuberculosis were produced by Streptomyces lividans as secreted extracellular proteins. An expression-secretion vector had been constructed that contained the promoter of xylanase A and the signal sequence of cellulase A. The latter contained two initiation codons preceded by a Shine-Dalgarno sequence plus eight nucleotides complementary to the 16S rRNA. The genes encoding the 38-kDa (Rv0934) and 19-kDa (Rv3763) proteins, respectively, were amplified by polymerase chain reaction and cloned into that vector. The recombinant proteins were then purified from the culture supernatants of the clones. The yields after purification were 80 mg/L for the 38-kDa protein and 200 mg/L for the 19-kDa protein. Sequence analysis of the N-terminal sequences showed a deletion of seven or eight amino acids for the 38-kDa protein, while in the 19 kDa protein 22 or 23 amino acids were lost, as compared with the respective wild type proteins. However, the 19 kDa recombinant protein had the same N-terminal sequence as the one recovered from the M. tuberculosis culture supernatant. The high yields obtained for these two proteins demonstrated the potential of S. lividans as an alternative host for the production of recombinant proteins from M. tuberculosis. The culture conditions have yet to be worked out to minimize proteolytic degradation and to recover intact products. PMID- 11888163 TI - Characterization of tdt genes for the degradation of tricyclic diterpenes by Pseudomonas diterpeniphila A19-6a. AB - Resin acids are tricyclic diterpenes that are toxic to aquatic life when released in high concentrations in pulp mill effluents. These naturally formed organic acids are readily degraded by bacteria and fungi; nevertheless, many of the mechanisms involved are still unknown. We report the localization, cloning, and sequencing of genes for abietane degradation (9.18 kb; designated tdt (tricyclic diterpene) LRSABCD) from the gamma-Proteobacterium Pseudomonas diterpeniphila A19 6a. Using gene knockout mutants, we demonstrate that tdtL, encoding a putative CoA ligase, is required for growth on abietic and dehydroabietic acids. A second gene knockout in tdtD, encoding a putative cytochrome P450 monooxygenase, reduced the growth of strain A19-6a on abietic and dehydroabietic acids as sole sources of carbon and energy, but did not eliminate growth. The degree of homology between P450TdtD and P450TerpC, the closest known P450 homologue to TdtD, identifies TdtD as a new member of the P450 superfamily. Hybridization of six of the tdt genes to genomic DNA of a related resin acid degrading bacterium Pseudomonas abietaniphila BKME-9 identified tdt homologues in this strain that utilizes aromatic ring dioxygenase genes (dit) to open the ring structure of abietic and dehydroabietic acids. These results suggest the tdt and dit genes may function in concert to allow these Pseudomonas strains to degrade resin acids. Homologues of several of the tdt genes were detected in resin acid degrading Ralstonia and Comamonas species within the beta- and gamma-Proteobacteria. PMID- 11888164 TI - Comparative degradation of oomycete, ascomycete, and basidiomycete cell walls by mycoparasitic and biocontrol fungi. AB - Fourteen fungi (primarily representing mycoparasitic and biocontrol fungi) were tested for their ability to grow on and degrade cell walls (CWs) of an oomycete (Pythium ultimum), ascomycete (Fusarium equisetii), and basidiomycete (Rhizoctonia solani), and their hydrolytic enzymes were characterized. Protein was detected in the cultural medium of eleven of the test isolates, and these fungi significantly degraded CWs over the 14-day duration of the experiment. In general, a greater level of CW degradation occurred for F. equisetii and P. ultimum than for R. solani. Fungi that degraded F. equisetii CWs were Coniothyrium minitans, Gliocladium roseum, Myrothecium verrucaria, Talaromyces flavus, and Trichoderma harzianum. Taxa degrading P ultimum CWs included Chaetomium globosum, Coniothyrium minitans, M. verrucaria, Seimatosporium sp., Talaromyces flavus, Trichoderma hamatum, Trichoderma harzianum, and Trichoderma viride. Production of extracellular protein was highly correlated with CW degradation. Considerable variation in the molecular weights of CW-degrading enzymes were detected among the test fungi and the CW substrates in zymogram electrophoresis. Multivariate analysis between CW degradation and hydrolysis of barley beta-glucan (beta1,3- and beta1,4-glucanases), laminarin (beta1,3- and beta1,6-glucanases), carboxymethyl cellulose (endo-beta1,4-glucanases), colloidal chitin (chitinases), and chitosan (chitosanases) was conducted. For F. equisetii CWs, the regression model accounted for 80% of the variability, and carboxymethyl cellulases acting together with beta-glucanases contributed an R2 of 0.52, whereas chitinases and beta-glucanases alone contributed an R2 of 0.11 and 0.12, respectively. Only 61% of the variability observed in the degradation of P. ultimum CWs was explained by the enzyme classes tested, and primarily beta glucanases (R2 of 0.53) and carboxymethyl cellulases (R2 of 0.08) alone contributed to CW break down. Too few of the test fungi degraded R. solani CWs to perform multivariate analysis effectively. This study identified several fungi that degraded ascomyceteous and oomyceteous, and to a lesser extent, basidiomycetous CWs. An array of enzymes were implicated in CW degradation. PMID- 11888165 TI - Glycine and alanine dehydrogenase activities are catalyzed by the same protein in Mycobacterium smegmatis: upregulation of both activities under microaerophilic adaptation. AB - Microaerophilic adaptation has been described as one of the in vitro dormancy models for tuberculosis. Studies on Mycobacterium tuberculosis adapted to low oxygen levels showed an enhancement of glycine dehydrogenase (deaminating) activity. We studied the physiology of the fast-growing, nonpathogenic strain of Mycobacterium smegmatis ATCC 607 under low oxygen by shifting the actively growing M. smegmatis cells to static microaerophilic growth conditions. This shifting of M. smegmatis culture resulted in a similar phenomenon as seen with M. tuberculosis, i.e., elevated glycine dehydrogenase activity. Further purification of glycine dehydrogenase from M. smegmatis demonstrated glyoxylate amination, but failed to demonstrate glycine deamination, even in the purified fraction. Moreover, the purified protein showed pyruvate amination as well as L-alanine deamination activities. By activity staining, the protein band positive for glyoxylate amination demonstrated only pyruvate amination in the presence of NAD. Absence of glycine deamination activity strongly suggested that alanine dehydrogenase of M. smegmatis was responsible for glyoxylate amination in the cell lysate. This was further confirmed by demonstrating the similar level of upregulation of both glyoxylate and pyruvate amination activities in the cell lysate of the adapted culture. PMID- 11888166 TI - Regulation of Raoultella terrigena comb.nov. phytase expression. AB - Phytases catalyze the release of phosphate from phytate (myo-inositol hexakisphosphate) to inositol polyphosphates. Raoultella terrigena comb.nov. phytase activity is known to increase markedly after cells reach the stationary phase. In this study, phytase activity measurements made on single batch cultures indicated that specific enzyme activity was subject to catabolite repression. Cyclic AMP (cAMP) showed a positive effect in expression during exponential growth and a negative effect during stationary phase. RpoS exhibited the opposite effect during both growth phases; the induction to stationary phase decreased twofold in the rpoS::Tn10 mutant, but the effect of RpoS was not clearly determined. Two phy::MudI1734 mutants, MW49 and MW52, were isolated. These formed small colonies in comparison with the MW25 parent strain when plated on Luria Bertani (LB) or LB supplemented with glucose. They did not grow in minimal media or under anaerobiosis, but did grow aerobically on LB and LB glucose at a lower rate than did MW25. The beta-galactosidase activity level in these mutants increased three to four fold during stationary growth in LB glucose and during anaerobiosis. Addition of cAMP during the exponential growth of MW52 on LB glucose provoked a decrease in beta-galactosidase activity during the stationary phase, confirming its negative effect on phytase expression during stationary growth. PMID- 11888167 TI - Estimation of vaginal probiotic lactobacilli growth parameters with the application of the Gompertz model. AB - Lactobacilli are widely described as probiotic microorganisms used to restore the ecological balance of different animal or human tracts. For their use as probiotics, bacteria must show certain characteristics or properties related to the ability of adherence to mucosae or epithelia or show inhibition against pathogenic microorganisms. It is of primary interest to obtain the highest biomass and viability of the selected microorganisms. In this report, the growth of seven vaginal lactobacilli strains in four different growth media and at several inoculum percentages was compared, and the values of growth parameters (lag phase time, maximum growth rate, maximum optical density) were obtained by applying the Gompertz model to the experimental data. The application and estimation of this model is discussed, and the evaluation of the growth parameters is analyzed to compare the growth conditions of lactobacilli. Thus, these results in lab experiments provide a basis for testing different culture conditions to determine the best conditions in which to grow the probiotic lactobacilli for technological applications. PMID- 11888168 TI - Proteolytic release of membrane-bound endo-(1,4)-beta-glucanase activity associated with cell wall softening in Achlya ambisexualis. AB - Branching and other cell wall softening events in fungi and oomycetes are thought to involve the activity of secreted enzymes, which are packaged in membrane vesicles and delivered to sites of cell expansion, there to work in a carefully regulated manner upon the structure of the wall. Here we demonstrate a latent endo-(1,4)-beta-glucanase activity in a mixed membrane fraction of the oomycete Achlya ambisexualis, which can be released by cysteine proteases with an increase of apparent activity. In addition, a similar endogenous process is strongly inhibited by the cysteine protease inhibitor iodoacetamide, while inhibitors of other types of proteases have a much smaller effect. Detergent treatment of membranes releases two glucanases detectable by electrophoretic activity staining, with apparent molecular masses of about 164 and 35 kDa. Proteolysis produces several activity bands, with major species having apparent molecular masses of about 149, 133, 48, 35, and 25 kDa. The ca. 35- and 25-kDa bands migrate in parallel with glucanases secreted during wall softening in vivo. We propose that the initiation of wall softening in Achlya involves the proteolytic processing and solubilization of at least some secreted endoglucanases. We also propose that the solubilization component of this process functions not just to provide the enzymes with access to wall matrix substrates but also may provide a mechanism for the eventual termination of their biological function. PMID- 11888169 TI - Cross-protection against challenge by intravenous Escherichia coli verocytotoxin 1 (VT1) in rabbits immunized with VT2 toxoid. AB - Rabbits challenged intravenously with Escherichia coli verocytotoxin (VT1, Shiga toxin 1, Stx1) die after developing diarrhea and paralysis, and this outcome can be prevented by pre-immunization with VT1 toxoid. In nonimmune rabbits, intravenously administered 125I-VT1 binds to the central nervous system and gastrointestinal tract, whereas in immunized animals, these organs are spared and the toxin localizes in the liver and spleen. In rabbits immunized with either VT1 or VT2 toxoids, both the homologous or heterologous toxins are prevented from binding to target organs. This has lead to the advancement of a hypothesis that cross-protection in vivo can be induced to both toxins by immunization with a toxoid even though these toxins do not exhibit cross-neutralization in vitro. It was shown that rabbits immunized with VT2 were fully protected from the intravenous administration of 10 LD50 and 50 LD50 of VT1, and this correlated directly with the protection from binding of this toxin to target organs. These findings have important implications on the design of the vaccination strategies to prevent human VT-mediated diseases and also validate the concept of testing for immunity to VT by monitoring the inhibition of binding of the 125I-VT to target organs in preference to performing LD50 assays. PMID- 11888170 TI - Tipping bucket mechanical errors and their influence on rainfall statistics and extremes. AB - Based on the error figures obtained after laboratory tests over a wide set of operational rain gauges from the network of the Liguria region, the bias introduced by systematic mechanical errors of tipping bucket rain gauges in the estimation of return periods and other statistics of rainfall extremes is quantified. An equivalent sample size is defined as a simple index that can be easily employed by practitioner engineers to measure the influence of systematic mechanical errors on common hydrological practice and the derived hydraulic engineering design. A few consequences of the presented results are discussed, with reference to data set reconstruction issues and the risk of introducing artificial climate trends in the observed rain records. PMID- 11888171 TI - Accounting for the spatial rainfall variability in urban modelling applications. AB - In hydrological and hydrodynamic modelling of urban catchments, the spatial variability of rainfall is often neglected. This spatial variability encloses two aspects: (1) the spatial variability of the statistical properties of rainfall, and (2) the non-uniform spatial distribution of rainfall over the modelled catchments. In an ongoing research project for the Ministry of the Flemish Community (Belgium), the influence of this spatial rainfall variability on the results of modelling applications is studied. At the same time, most efficient methods to reduce this influence are determined. The results of the research can be applied directly in Flanders. They consist of a combination of unified IDF relationships, spatial correction factors (generally applicable formulas), a stochastic simulation model for spatial rainfall (software) and a methodology for improving the spatial correction factors in a case-specific way by performing simulations with the model. PMID- 11888172 TI - Development of an internet-based rainfall atlas for Alabama. AB - In the United States, rainfall information needed for intensity-duration frequency (IDF) curves or design storm hyetographs can be found in TP 40, HYDRO 35, and the NOAA Atlas 2. Additional rainfall data collected since the dates of those publications, and improved methods for statistical treatment of data, have motivated update studies in several regions of the United States. One of the new studies has been performed for the State of Alabama. Results of the Alabama study are embodied in an internet-based graphical user interface, which permits users to interactively point and click on a geographical location of interest, and have IDF curves and/or storm hyetographs returned on demand. Reactions to the internet based rainfall atlas have been promising, and have led to additional work for the National Weather Service, Office of Hydrologic Development. PMID- 11888173 TI - Stochastic generation and disaggregation of hourly rainfall series for continuous hydrological modelling and flood control reservoir design. AB - In the urban environment, stormwater detention basins are a powerful means to limit the frequency of sewer system failures and consecutive urban flooding. To design such waterworks or to check their efficiency, it is possible to carry out continuous rainfall-runoff modelling. A long-term discharge series obtained from a long-term rainfall series is used as input for a storage model describing the detention basin behaviour: the basin behaviour may be consequently studied over a long period. The provided statistical information on the working state frequency, failure frequency, ... of the detention basin is of high interest for the basin diagnostic or for its design. This paper presents the whole methodology which leads to production of such statistical information and especially: the models used to generate long term rainfall series with a short time step, the rainfall runoff model used to transform the later series into a long term discharge series, and the model used to describe the behaviour of the detention basin. This methodology was applied to evaluate the efficiency of 4 detention basins built for stormwater control and flood mitigation. They are situated on a Swiss urban catchment (Chamberonne catchment--40 km2) collecting water from the Mebre and Sorge rivers. PMID- 11888174 TI - Short-term risk forecasts of heavy rainfall. AB - Methodologies for risk forecasts of severe weather hardly exist on the scale of nowcasting (0-3 hours). Here we discuss short-term risk forecasts of heavy precipitation associated with local thunderstorms. We use COTREC/RainCast: a procedure to extrapolate radar images into the near future. An error density function is defined using the estimated error of location of the extrapolated radar patterns. The radar forecast is folded ("smeared") with the density function, leading to a probability distribution of radar intensities. An algorithm to convert the radar intensities into values of precipitation intensity provides the desired probability (or risk) of heavy rainfall at any position within the considered window in space and time. We discuss, as an example, a flood event from summer 2000. PMID- 11888176 TI - X-Band local area weather radar--preliminary calibration results. AB - DHI has developed a cost-effective X-Band Local Area Weather Radar (LAWR) with a typical range (radius) of 60 km, 500 x 500 m areal resolution and 253 reflection levels. The development is performed in a co-operation with a number of European partners, including Danish Meteorological Institute. The specifications of the weather radar and preliminary results from the calibration are presented. Good calibration results have been obtained using high-resolution rain gauges. PMID- 11888175 TI - Measurement, calibration of rainfall-runoff models and assessment of the return period of flooding events at urban catchment Kumodraz in Belgrade. AB - For the purpose of re-design and improvement of the combined sewer system at the Kumodraz catchment in Belgrade, measurements of rainfall and runoff at the catchment were established in late 1997. Observed data are used for calibration and validation of two rainfall-runoff models: the detailed model BEMUS (Belgrade Model) and a conceptual hydrologic model Visual OTTHYMO. The major facilities of the recommended solution for re-design of the existing system are three retention ponds and outlet into a trunk. The paper briefly presents assessment of design flows for these four locations of the catchment based on results from the calibrated models. Data on intense storms that caused severe flooding within the catchment (and also all over Belgrade and other parts of Serbia) in June and July 1999 are used for comparison of design flows with a real event. During these storms the flow gauging equipment at the catchment was destroyed and only the rainfall data was available. In order to analyze the system under extreme conditions it is therefore necessary to perform hydrograph simulations. The basic idea was to check return periods of rainfall and runoff for these storms. PMID- 11888177 TI - WMO questionnaire on recording precipitation gauges: state-of-the-art. World Meteorological Organization. AB - To receive information on recording precipitation gauges used operationally by national meteorological services the World Meteorological Organization started a worldwide inquiry. This included the type of recording precipitation gauge (RPG), their manufacturer, measuring system applied, number of RPGs used, methods of recording and data transmission, orifice area size and installation heights, heating and windshield application as well as the question on the need to organising global intercomparisons of RPG measurements. The results of the 118 responses received (out of 180 countries) are presented and discussed. They show that there is a great variety in instruments and methods of observation of precipitation intensity used not only worldwide but also in the same country. This variety exceeds by far the variety in manual standard precipitation gauges. PMID- 11888178 TI - An enquiry into rainfall data measurement and processing for model use in urban hydrology. AB - Rain data are collected all over the world because water is of paramount importance to all human life. WMO has provided standards for collection and standardized data processing of daily rainfall measurements. Currently no such standards are available for gauges with a resolution suitable for urban hydrology, where the resolution in time must not exceed a few minutes. The Group on Urban Rainfall under the International Water Association has made a comparison of national standards by means of a survey of 77 questions sent to 44 countries. The paper discusses the first results of the answers of the survey. Currently tipping bucket gauges are the dominating method of obtaining high resolution rain data, but the numbers of weighing gauges and radar measurements are rapidly growing. It is necessary to try to increase the awareness of documentation of current standards and to agree on standards for measurements and data processing on an international level in the future. PMID- 11888179 TI - Management of a detention-settling basin using radar data and risk notion. AB - This paper presents a practical application of sewage network management strategy using radar data to control a detention-settling basin. This practical application, used in operational mode since January 2000, is based on three main notions. Firstly, the use of radar data without rainfall estimation. Secondly, the definition of some gradual risk levels for the sewage network by a detailed modelling of the sewage system functioning. Thirdly the definition of relations between risk levels for the sewage network and types of rain events defined from radar data. The operational application produces gradual alarms for decision making assistance: no risk, potential risk and confirmed risk. PMID- 11888180 TI - Comparison of rain gauge and radar data as input to an urban rainfall-runoff model. AB - This paper presents an application of radar data (DX-product of the German Weather Service) with a high resolution in space (1 degree x 1 km) and time (delta t = 5 minutes) in urban hydrology. The radar data and data of rain gauges with different locations in the test catchment were compared concerning their suitability as input into an urban rainfall-runoff model. In order to evaluate the accuracy of model simulation results, five evaluation criteria have been specified which are relevant for an efficient management of sewer systems and wastewater treatment plants. The results demonstrate that radar data should be used in urban hydrology if distances > 4 km between rain gauge and catchment exist and for catchments with a density of rain gauges smaller than 1 rain gauge per 16 km2. PMID- 11888181 TI - On the properties of stochastic intermittency in rainfall processes. AB - In this work we propose a mixed approach to deal with the modelling of rainfall events, based on the analysis of geometrical and statistical properties of rain intermittency in time, combined with the predictability power derived from the analysis of no-rain periods distribution and from the binary decomposition of the rain signal. Some recent hypotheses on the nature of rain intermittency are reviewed too. In particular, the internal intermittent structure of a high resolution pluviometric time series covering one decade and recorded at the tipping bucket station of the University of Genova is analysed, by separating the internal intermittency of rainfall events from the inter-arrival process through a simple geometrical filtering procedure. In this way it is possible to associate no-rain intervals with a probability distribution both in virtue of their position within the event and their percentage. From this analysis, an invariant probability distribution for the no-rain periods within the events is obtained at different aggregation levels and its satisfactory agreement with a typical extreme value distribution is shown. PMID- 11888182 TI - The effect of topography, season and weather situation on daily precipitation gradients in 60 Swiss valleys. AB - Daily precipitation in the Swiss valleys decreases frequently with altitude, particularly in southern Switzerland. Only one-third of precipitation days show positive precipitation gradients with a fairly strong correlation of precipitation with altitude (R2 > or = 0.5). Generally, precipitation gradients are larger in the north than in the south but there are large differences among valleys situated near to each other. The effects of precipitation amount, weather situation, region and season, wind speed and topography were not very significant at all. It seems that the positioning of the upper and lower precipitation gauges under specific topographic and climate conditions can affect the precipitation gradients in a particular valley. PMID- 11888183 TI - Investigating the changes in extreme rainfall series recorded in an urbanised area. AB - The aim of this study is to investigate and quantify changes in the rainfall regime of the metropolitan area of Palermo characterised by increasingly strong urbanisation. The rainfall data, considered in this study, were collected on a yearly basis from eight rain gauges within and outside the metropolitan area of Palermo, Sicily, Italy. A preliminary analysis made on the annual total rainfall depths showed a global reduction of total annual rainfall, with two different trends: more regular for the series observed in the rain gauges within the urbanised area and more variable for the series observed in the rain gauges outside the area. A further analysis has been performed using the series of maximum intensity for fixed duration (1, 3, 6, 12, 24 hrs) and annual daily maxima. The analysis of the trend in the extreme rainfall series has been performed by estimating the maximum rainfall depth corresponding to a fixed return period using the EV1 distribution with parameters estimated using L moments. The analysis of all series indicates a global reduction of rainfall intensities, both for internal and external series, in disagreement with the results obtained by other authors. PMID- 11888185 TI - Dissemination of regional rainfall analysis in design and analysis of urban drainage at un-gauged locations. AB - A research program in Denmark on statistical modelling of rainfall has resulted in a model for regional distribution of rainfall extremes. The results show that extreme rainfalls critical to the hydraulic function of urban drainage systems and the pollution discharge are subject to a significant regional variation of extreme rainfalls throughout the country. This has implications for design and analysis of all practical problems related to urban drainage, since the rainfall data so far recommended as input to engineering analyses underestimates the problems. Consequently, the Danish Water Pollution Control Committee has issued a statement recommending a new engineering practice. The dissemination of the research results proved to be difficult due to lack of understanding of the concepts of the new paradigm by practitioners. The traditional means of communication was supplemented by user-friendly spreadsheets and easy access to rainfall data as well as giving courses on the new paradigm. This has eased the implementation of the new concepts greatly. PMID- 11888184 TI - 100 years of Belgian rainfall: are there trends? AB - In 1999 the digitisation of old rainfall records of measurements at Uccle (Belgium) was completed, which resulted in a unique rainfall series of 100 years (period 1898-1997). This is an ideal opportunity to search for trends in the rainfall over the last century. Large variations in rainfall probability over the century have been observed. For small aggregation levels there is a small decrease in extreme rainfall events over the century. For large aggregation levels there is a more explicit increase in extreme rainfall. Because the rainfall on seasonal aggregation level is only slightly increased, the increase in extreme rainfall events for aggregation levels between a few days and a few months can only occur due to larger clustering. However, the final conclusion is that no significant trend can be observed. A pure random variation of the rainfall can cause equally large variations. This does not exclude a possible trend in flooding frequency, due to the strong increase in urbanisation over the last century. PMID- 11888186 TI - IDAF (intensity-duration-area frequency) curves of extreme storm rainfall: a scaling approach. AB - Intensity-duration-area frequency curves, IDAF, are determined for the evaluation of design storms using a scaling approach. The variability of maximum annual rainfall intensity in area and duration is represented through the scaling properties in time and space. Thus the scaling relationships of mean rainfall intensity with area and duration are derived using the concepts of dynamic scaling and statistical self-affinity. For a lognormal distribution of rainfall intensity a multiscaling lognormal model is obtained. This gives the IDAFcurves of extreme storm rainfall. An application is made to the metropolitan area of Milano. PMID- 11888187 TI - A Markov modulated Poisson process model for rainfall increments. AB - The problems encountered when using traditional rectangular pulse hierarchical point process models for fine temporal resolution and the growing number of available tip-time records suggest that rainfall increments from tipping-bucket gauges be modelled directly. Poisson processes are used with an arrival rate modulated by a Markov chain in continuous time. The paper shows how, by using two or three states for this chain, much of the structure of the rainfall intensity distribution and the wet/dry sequences can be represented for time-scales as small as 5 minutes. PMID- 11888188 TI - Evaluation of an artificial neural network rainfall disaggregation model. AB - Previous research produced an artificial neural network (ANN) temporal rainfall disaggregation model. After proper training the model can disaggregate hourly rainfall records into sub-hourly time increments. In this paper we present results from continued evaluations of the performance of the ANN model specifically examining how the errors in the disaggregated rainfall hyetograph translate to errors in the prediction of the runoff hydrograph. Using a rainfall runoff model of a hypothetical watershed we compare the runoff hydrographs produced by the ANN-predicted 15-minute increment rainfall pattern to runoff hydrographs produced by (1) the observed 15-minute increment rainfall pattern, (2) the observed hourly-increment rainfall pattern, and (3) the 15-minute increment rainfall pattern produced by a disaggregation model based on geometric similarity. For 98 test storms the peak discharges produced by the ANN model rainfall pattern had a median under-prediction of 16.6%. This relative error was less than the median under-prediction in peak discharge when using the observed 15-minute rainfall patterns aggregated to hourly increments (40.8%), and when using rainfall patterns produced by the geometric similarity rainfall disaggregation model (21.9%). PMID- 11888189 TI - Evolution of fracture care of distal femoral fractures. PMID- 11888190 TI - Evolution of minimally invasive plate osteosynthesis (MIPO) in the femur. AB - Problems with conventional open reduction and internal plate fixation of distal femoral fractures are well established. These problems have been associated with extensile exposures of the fracture site. "Biological plating", like intramedullary nail fixation, of distal femoral fractures preserves the soft tissues about the fracture, and is associated with early fracture consolidation and low rates of infection. Anatomical restoration of the articular surface continues to be the main goal in the treatment of these fractures regardless of the stabilization technique. Submuscular plating techniques, which provide for closed reduction of the diaphyseal/metaphyseal component of the fracture, have improved significantly. PMID- 11888191 TI - The development of the distal femur Less Invasive Stabilization System (LISS). AB - Fractures around the knee typically require operative fixation to achieve an acceptable, functional outcome. The idea behind the Less Invasive Stabilization System (LISS) was to combine the advantages of both interlocked intramedullary nailing techniques and the early advances of the so-called biological plating technique into one system. This paper introduces the mechanical concept of a locked internal fixator and details some important aspects of the anatomical and biomechanical development of the LISS. PMID- 11888192 TI - Distal femoral fractures: current treatment, results and problems. AB - The evolution of treatment for supracondylar femoral fractures has sequentially addressed the difficulties of alignment, articular reduction, stabilization and fracture union. Adequate surgical stabilization and early motion diminished stiffness, while newer indirect techniques in handling periarticular tissues have greatly improved union rates. Indirect methods of reduction require an understanding of anatomy and deformity to avoid malalignment. The problems we currently face are fixation in osteoporotic bone or small distal articular segments. PMID- 11888193 TI - Distal femoral fracture fixation utilizing the Less Invasive Stabilization System (L.I.S.S.): the technique and early results. AB - The treatment of supracondylar femoral fractures in the past three decades has evolved from non-operative to operative treatment. While operative fixation utilizing either plate fixation or rigid intramedullary nail fixation has improved patient outcomes, the problems of malunion, nonunion, need for bone grafting, joint stiffness, and infection persist. An emphasis on maintenance of the soft tissue envelope around fractures has improved efficacy in increasing osseous healing and decreasing infection. Out of this movement grew the concept of submuscular plating for distal femoral fractures, and subsequently L.I.S.S. fixation (Less Invasive Stabilization System) for distal femoral fractures. The technique and early results utilizing the L.I.S.S. for distal femoral fractures is described. The technique of L.I.S.S. fixation first begins with traditional direct visualization and internal fixation of the articular surface. Closed reduction is then performed on the metaphyseal / diaphyseal component of the fracture, followed by submuscular fixation utilizing the L.I.S.S. fixation. The L.I.S.S. can best be thought as an "internal" external fixator. PMID- 11888194 TI - Minimally invasive fracture stabilization of distal femoral fractures with the LISS: a prospective multicenter study. Results of a clinical study with special emphasis on difficult cases. AB - The LISS-DF (Less invasive stabilization system-distal femur) is a new type of implant system for the treatment of distal femoral fractures according to the principles of "Minimally Invasive Surgery". A plate, pre-contoured to the anatomy, is inserted through a minimally invasive incision into the epiperiosteal space by means of an aiming device after indirect, closed fracture reduction. The implant is stabilized by insertion of screws which lock into the plate holes and prevent tilting. This is performed with the aid of an aiming device and through stab incisions. It is not necessary for a large area to be exposed at the fracture site. As part of an AO prospective multicenter study, the new system was applied to 112 patients with 116 fractures. The time to follow-up was on average 13.7 months (minimum 7 months, maximum 33 months). Fractures treated were distal femoral shaft and supracondylar femoral fractures. Eight patients died during the study of causes unrelated to the implant. Of the remaining 104 patients with 107 fractures, 96 patients with 99 fractures were available for complete follow-up (93% follow-up rate). In 90% of all cases treated and followed up, the fracture had consolidated during the period of observation. Twenty-three revision operations were necessary in 21 patients. In two cases, implant failure occurred as the result of a pseudarthrosis. The complications can be attributed in nearly all cases to the severity of the trauma and/or a lack of experience when applying the new style implant to a wider range of indications. The results of the study show that with a sound knowledge of the operative technique and careful preoperative planning this system represents an excellent, safe procedure for the treatment of almost all distal femoral fracture types including periprosthetic fractures of the distal femur. There is generally no need for primary cancellous bone grafting. PMID- 11888195 TI - Distal femoral fractures and LISS stabilization. AB - In recent years, the technique of surgical stabilization in the distal femur has changed. This change decreased the number of non unions and the need for bone grafting. Minimally invasive surgical techniques with a submuscular plate placement have replaced the emphasis on anatomical reduction in the shaft area. Reconstruction of complex articular injuries has been simplified by more direct visualization of the articular surface with the lateral peripatellar approach. Problems remaining are surgical technique and implant considerations. The Less Invasive Stabilization System (LISS) simplifies the surgical technique for percutaneous plate osteosynthesis. An insertion guide is used to insert monocortical, self-tapping screws through a stab incision. A thread in the plate provides the angular stability for the anchoring of these screws. In extra articular fractures and simple intra-articular fractures, the distal femoral nail permits intramedullary stabilization. A spiral blade improves fixation of the distal femoral condylar block. Despite the enhanced surgical technique and implant possibilities, a great number of patients show a functional deficiency. These are particularly patients with complex intra-articular fractures. The 'fatigue failure' of the osteoporotic implant-bone construct is a problem in elderly patients. The LISS represents a good option to avoid the addition of bone cement to an osteosynthesis. PMID- 11888196 TI - Fixation of distal femoral fractures above total knee arthroplasty utilizing the Less Invasive Stabilization System (L.I.S.S.). AB - Supracondylar femoral fractures above total knee arthroplasty remain a treatment challenge. Complication rates as high as 30% are associated with both nonoperative and operative treatment. Conventional plate fixation and rigid intramedullary nail fixation has improved the treatment of these fractures. However, problems still exist in the setting of a short distal femoral block and/or significant osteoporosis. Less Invasive Stabilization System (L.I.S.S.) fixation has been utilized for the treatment of supracondylar femoral fractures above total knee arthroplasty. Multiple fixed angle screws give optimal fixation around the femoral component. Advantages appear to include maintenance of distal femoral fixation, low infection, and low need for bone grafting. PMID- 11888197 TI - Unfolding of triosephosphate isomerase from Trypanosoma brucei: identification of intermediates and insight into the denaturation pathway using tryptophan mutants. AB - The unfolding of triosephosphate isomerase (TIM) from Trypanosoma brucei (TbTIM) induced by guanidine hydrochloride (GdnHCl) was characterized. In contrast to other TIMs, where unfolding is a two or three state process, TbTIM showed two intermediates. The solvent exposure of different regions of the protein in the unfolding process was characterized spectroscopically with mutant proteins in which tryptophans (W) were changed to phenlylalanines (F). The midpoints of the transitions measured by circular dichroism, intrinsic fluorescence, and catalytic activity, as well as the increase in 1-aniline 8-naphthalene sulfonate fluorescence, show that the native state was destabilized in the W12F and W12F/W193F mutants, relative to the wild-type enzyme. Using the hydrodynamic profile for the unfolding of a monomeric TbTIM mutant (RMM0-1TIM) measured by size-exclusion chromatography as a standard, we determined the association state of these intermediates: D*, a partially expanded dimer, and M*, a partially expanded monomeric intermediate. High-molecular-weight aggregates were also detected. At concentrations over 2.0 M GdnHCl, the hydrodynamic properties of TbTIM and RMM0-1TIM are the same, suggesting that the dimeric intermediate dissociates and the unfolding proceeds through the denaturation of an expanded monomeric intermediate. The analysis of the denaturation process of the TbTIM mutants suggests a sequence for the gradual exposure of W residues: initially the expansion of the native dimer to form D* affects the environments of W12 and W159. The dissociation of D* to M* and further unfolding of M* to U induces the exposure of W170. The role of protein concentration in the formation of intermediates and aggregates is discussed considering the irreversibility of this unfolding process. PMID- 11888198 TI - Trisamine C(60)-fullerene adducts inhibit neuronal nitric oxide synthase by acting as highly potent calmodulin antagonists. AB - C(60)-Fullerene trisamine adducts inhibit neuronal nitric oxide synthase and calcineurin phosphatase activities in a manner completely reversible by calmodulin. As measured by difference spectroscopy, D(3)-trisamine and C(3) semiamine fullerene adducts displace trifluoperazine bound to calmodulin coincident with their binding. These binding events are complete at a molar ratio of 4 mol added fullerene per mole calmodulin. Trisamine fullerene adducts alter the native electrophoretic mobility of calmodulin, producing a heterogeneity of bands with associated fullerene. D(3)- and C(3)-trisamine fullerene adducts interact with dansylated calmodulin, producing a 50% loss of maximal fluorescence at concentrations of 30 nM. At higher concentrations than those required to inhibit neuronal nitric oxide synthase, trisamine fullerene adducts inhibit nitric oxide formation by the cytokine-inducible nitric oxide synthase isoform. These inhibitions are fully reversible by calmodulin and skeletal muscle troponin C but not by skeletal muscle parvalbumin. Of the trisamine fullerene adducts tested only the C(3)- and D(3)-semiamine adducts inhibit Ca(2+)-dependent nitric oxide production in GH(3) pituitary cells. These observations support the proposal that trisamine C(60)-fullerene adducts are potent calmodulin antagonists, some of which display activity in intact cellular systems. PMID- 11888199 TI - Refined structure and metal binding site of the kalata B1 peptide. AB - The cyclic polypeptide kalata B1 from the African plant Oldenlandia affinis DC consists of 29 amino acid residues with three disulfide linkages. In this study we used two-dimensional NMR spectroscopy to investigate the three-dimensional structure of the peptide and to determine the disulfide connectivities. Nuclear Overhauser effects (NOEs) between neighboring beta-protons of the cysteines detected at 750 MHz provided evidence for the disulfide connectivity pattern 5 13, 17-29, and 22-27. These disulfide linkages were confirmed by three dimensional structures calculated from input constraints derived solely from NOEs without explicit disulfide connectivities. Kalata B1 is insoluble in aqueous solution above pH 3.5, but in a 50-50 water-methanol mixture, it was possible to use natural abundance two-dimensional (15)N-(1)H heteronuclear single quantum coherence spectroscopy to study the hydrophobic peptide from pH 2 to 10. The addition of methanol resulted in no significant structural changes. Although the peptide contains three prolyl residues, no evidence of multiple conformers was detected at any pH. The addition of Mn(2+) to kalata B1 resulted in selective broadening of resonances from Asn 23, Thr 24, and Glu 15; these results suggest that these three residues are involved in a specific metal binding site. PMID- 11888200 TI - Amino acid sequence and glycosylation of functional unit RtH2-e from Rapana thomasiana (gastropod) hemocyanin. AB - The complete amino acid sequence of Rapana thomasiana hemocyanin functional unit RtH2-e was determined by direct sequencing and matrix-assisted laser desorption ionization mass spectrometry of peptides obtained by cleavage with EndoLysC proteinase, chymotrypsin, and trypsin. The single-polypeptide chain of RtH2-e consists of 413 amino acid residues and contains two consensus sequences NXS/T (positions 11-19 and 127-129), potential sites for N-glycosylation. Monosaccharide analysis of RtH2-e revealed a carbohydrate content of about 1.1% and the presence of xylose, fucose, mannose, and N-acetylglucosamine, demonstrating that only N-linked carbohydrate chains of high-mannose type seem to be present. On basis of the monosaccharide composition and MALDI-MS analysis of native and PNGase-F-treated chymotryptic glycopeptide fragment of RtH2-e the oligosaccharide Man(5)GlcNAc(2), attached to Asn(127), is suggested. Multiple sequence alignments with other molluscan hemocyanin e functional units revealed an identity of 63% to the cephalopod Octopus dofleini and of 69% to the gastropod Haliotis tuberculata. The present results are discussed in view of the recently determined X-ray structure of the functional unit g of the O. dofleini hemocyanin. PMID- 11888201 TI - Mapping of the chick heme oxygenase-1 proximal promoter for responsiveness to metalloporphyrins. AB - Heme oxygenase (HO) catalyzes the rate-controlling step of physiologic heme catabolism, namely, the oxidation of the alpha-methene bridge of the macrocycle with formation of CO, Fe, and biliverdin. HO-1, the first isoform of HO to be identified, is highly inducible by a large number of physical and chemical factors. Many of these factors cause oxidative or other stresses to cells. In this work, we have studied the regulation of the chick HO-1 gene, using selected promoter--reporter constructs of the gene transiently or stably transfected into primary cultures of chick embryo liver cells or into the LMH line of chicken hepatoma cells. By use of deletional and mutational analyses, DNase protection, and electromobility shift DNA-binding assays, we identified a heretofore undefined regulatory region in the 5'-UTR of the chick HO-1 gene which confers up regulation of reporter gene (luciferase) expression in the presence of heme and other selected metalloporphyrins. This new metalloporphyrin-responsive element (MPRE) was localized to a 200-bp region 3.8 to 3.6 kb upstream of the transcription starting point of the chick HO-1 gene. It responded particularly to heme and cobalt protoporphyrin with maximal inductions at 10-15 microM concentrations and 15-18 h of exposure. In contrast, sodium arsenite, a prototypical stress-type inducer of HO-1, led to down-regulation of the reporter gene down stream of MPRE. DNase analysis identified an 18-mer oligonucleotide that was required for the metalloporphyrin response (5'-(-3711)TATTGCAGCTGTGTGGGG 3'). Mutations at any of four sites within this oligonucleotide abrogated the metalloporphyrin-dependent up-regulation of reporter gene expression. Nuclear protein extracts of cells treated with heme or cobalt protoporphyrin showed specific enhanced binding to this 18-mer. We conclude that the chick HO-1 promoter region contains a unique sequence that subserves up-regulation of the gene by metalloporphyrins and propose the name "metalloporphyrin-responsive element" for this sequence. PMID- 11888202 TI - Aluminum affects membrane physical properties in human neuroblastoma (IMR-32) cells both before and after differentiation. AB - The capacity of Al(3+) to induce changes in the physical properties of plasma membrane from human neuroblastoma cells (IMR-32) was investigated, and the magnitude of the changes was compared with that obtained after cell differentiation to a neuronal phenotype. Similarly to our previous results in liposomes, Al(3+) (10 to 100 microM) caused a significant loss of membrane fluidity, being the differentiated cells more affected than the nondifferentiated cells. Al(3+) also increased the relative content of lipids in gel phase and promoted lipid rearrangement through lateral phase separation, with the magnitude of this effect being similar in nondifferentiated and differentiated cells. Since membrane physical properties depend on bilayer composition, we characterized the content of proteins, phospholipids, cholesterol, and fatty acids in the IMR-32 cells before and after differentiation. Differentiated cells had a significantly higher content of unsaturated fatty acids, creating an environment that favors Al(3+)-mediated effects on the bilayer fluidity. The neurotoxic effects of Al(3+) may be, at least in part, due to alterations of neuronal membrane physical properties, with potential consequences on the normal functioning of membrane related cellular processes. PMID- 11888203 TI - Glycation-induced matrix stability in the rabbit achilles tendon. AB - Connective tissue susceptibility to nonenzymatic glycation was examined following 0, 2, 4, 6, 8, and 10 weeks of incubating the rabbit Achilles tendon in phosphate buffered saline containing ribose (glycated). The biomechanical integrity of the glycated tendons was then compared to control tendons incubated in phosphate buffered saline (non-glycated) at each time interval, while the biochemical stability of both groups of tendons was determined by examining collagen extractability and the formation of pentosidine at 8 weeks. Whereas there were no significant biomechanical differences between control and glycated tendons at 0- and 2-week intervals (P > 0.05), moderately significant increases in maximum load, energy to yield, and toughness of glycated tendons were observed at 4 weeks. Beyond 4 weeks of incubation, the differences between glycated and non glycated tendons became highly significant, as glycated tendons withstood more load and tensile stress (P < 0.01 for each variable), attained significantly higher modulus of elasticity (P < 0.01), absorbed more energy (P < 0.01), and became tougher (P < 0.01) than controls. These differences in the biomechanical indices of the effects of glycation were stable between the 6th and 10th week of glycation. The maximum increases in the biomechanical measurements as a result of glycation were 29% for maximum load, 125% for stress, 19% for strain, 106% for Young's modulus of elasticity, 14% for energy to yield, and 57% for toughness. Biochemical analysis showed a 61% reduction in the extractability of neutral salt soluble collagen, a 48% decrease in acid-soluble collagen, and a 29% decline in pepsin-soluble collagen in glycated tendons (P < 0.01). In contrast, there was a 28% increase in the amount of insoluble collagen and significantly higher amounts of pentosidine (P < 0.01) in glycated tendons. Collectively, these biomechanical and biochemical results suggest that nonenzymatic glycation may explain the altered stability of connective tissue matrix induced by the processes of diabetes and aging. PMID- 11888204 TI - Nonspecific deadenylation on sarcin/ricin domain RNA catalyzed by gelonin under acidic conditions. AB - Gelonin is a single-chain ribosome-inactivating protein that can hydrolyze the glycosidic bond of a highly conserved adenosine residue in the sarcin/ricin domain (SRD) of the largest RNA in ribosome and thus irreversibly inhibit protein synthesis. Recently, the specificity in substrate recognition was challenged by the fact that gelonin could remove adenines from some other oligoribonucleotide substrates. However, the site specificity of gelonin to deadenylate various substrates were unknown. Hereby, the effect of pH values upon site specificity of the deadenylation activity of gelonin was studied using the synthetic oligoribonucleotide (named SRD RNA) that mimicked the ribosomal SRD. Interestingly, gelonin gradually acquired the ability to nonspecifically remove adenines from SRD RNA when pH values changed from neutral to acidic conditions. Another two SRD RNA mutants, either with the conserved adenosine deleted or with the tetraloop converted, showed very similar cleavage style to wild-type SRD RNA, underscoring the important role of pH value in site specificity of recognition by gelonin. Furthermore, the RNA N-glycosidase activity of gelonin was also enhanced with the decreasing of pH values. In addition, no obvious change was observed in the molecular conformation of gelonin at various pH values. Taken together, our data implied that the protonation of adenosines in SRD RNA was potentially an important factor for the nonspecific deadenlyation by gelonin. PMID- 11888205 TI - Investigation of the active site of the extracellular beta-D-xylosidase from Aspergillus carbonarius. AB - The catalytic amino acid residues of the extracellular beta-D-xylosidase (beta-D xyloside xylohydrolase, EC 3.2.1.37) from Aspergillus carbonarius was investigated by the pH dependence of reaction kinetic parameters and chemical modifications of the enzyme. The pH dependence curves gave apparent pK values of 2.7 and 6.4 for the free enzyme, while pK value of 4.0 was obtained for the enzyme-substrate complex using p-nitrophenyl beta-D-xyloside as a substrate. These results suggested that a carboxylate group and a protonated group- presumably a histidine residue--took part in the binding of the substrate but only a carboxylate group was essential in the substrate cleavage. Carbodiimide- and Woodward's reagent K-mediated chemical modifications of the enzyme also supported that a carboxylate residue, located in the active center, was fundamental in the catalysis. The pH dependence of inactivation revealed the involvement of a group with pK value of 4.4, proving that a carboxylate residue relevant for hydrolysis was modified. During modification V(max) decreased to 10% of that of the unmodified enzyme and K(m) remained unchanged, supporting that the modified carboxylate group participated in the cleavage and not in the binding of the substrate. We synthesized and tested a new, potential affinity label, N bromoacetyl-beta-d-xylopyranosylamine for beta-D-xylosidase. The A. carbonarius beta-D-xylosidase was irreversible inactivated by N-bromoacetyl-beta-D xylopyranosylamine. The competitive inhibitor beta-D-xylopyranosyl azide protected the enzyme from inactivation proving that the inactivation took place in the active center. Kinetic analysis indicated that one molecule of reagent was necessary for inactivation of one molecule of the enzyme. PMID- 11888206 TI - Expression, preparation, and high-throughput screening of caspase-8: discovery of redox-based and steroid diacid inhibition. AB - Because of the intimate role of caspase-8 in apoptosis signaling pathways from FAS, TNFR1, and other death receptors, the enzyme is a potentially important therapeutic target. We have generated an Escherichia coli expression construct for caspase-8 in which a His-tag sequence is inserted ahead of codon 217 of caspase-8. The strain produced a significant amount of soluble His-tagged 31-kDa inactive single-chain enzyme precursor. This 31-kDa protein could be purified to 98% purity. Hydroxyapatite resolved the enzyme into two species, one with the appropriate 31,090 relative mass and the other with 178 units additional mass. The latter proved to result from E. coli-based modification of the His-tag with one equivalent of glucono-1,5-lactone. The purified proteins could be activated by autoproteolysis to the appropriate 19- plus 11-kDa enzyme by the addition of dithiothreitol in appropriate buffer conditions. This yielded an enzyme with specific activity of 4-5 units/mg against 200 microM Ac-IETD-pNA at 25 degrees C. The fully active protein was used in a high-throughput screen for inhibitors of caspase-8. A preliminary robustness screen demonstrated that caspase-8 is susceptible to reactive oxygen-based inactivation in the presence of dithiothreitol (DTT) but not in the presence of cysteine. Investigation into the mechanism of this inactivation showed that quinone-like compounds were reduced by DTT establishing a reactive oxygen generating redox cycle the products of which (likely H(2)O(2)) inactivated the enzyme. A new class of caspase-8 inhibitors, steroid-derived diacids, with affinity in the low micromolar range were discovered in the refined screen. Structure--activity investigation of the inhibitors showed that both the steroid template and the acid moieties were required for activity. PMID- 11888207 TI - Glucose uptake and lactate production in cells exposed to CoCl(2) and in cells overexpressing the Glut-1 glucose transporter. AB - Glut-1-mediated glucose transport is augmented in response to a variety of conditions and stimuli. In this study we examined the metabolic fate of glucose in cells in which glucose transport is stimulated by exposure to CoCl(2), an agent that stimulates the expression of a set of hypoxia-responsive genes including several glycolytic enzymes and the Glut-1 glucose transporter. Similarly, we determined the metabolic fate of glucose in stably transfected cells overexpressing Glut-1. Exposure of Clone 9 liver cell line, 3T3-L1 fibroblasts, and C(2)C(12) myoblasts to CoCl(2) resulted in an increase glucose uptake and in the activity of glucose phosphorylation ("hexokinase") and lactate dehydrogenase. In cells treated with CoCl(2), the net increase in glucose taken up was accounted for by its near-complete conversion to lactate. Cells stably transfected to overexpress Glut-1 also exhibited enhanced net uptake of glucose with the near-complete conversion of the increased glucose taken up to lactate; however, the effect in these cells was observed in the absence of any change in the activity of two glycolytic enzymes examined. These findings suggest that in cells in which glucose transport is rate-limiting for glucose metabolism, enhancement of the glucose entry step per se results in a near-complete conversion of the extra glucose to lactate. PMID- 11888208 TI - Identification of novel E2F1-regulated genes by microarray. AB - The E2F pathway has been proposed to regulate genes involved in the transition from quiescence into DNA synthesis. However, this hypothesis has not been rigorously tested on a genomic scale. Toward this end, we have infected quiescent mouse fibroblasts, which do not express E2F1, with an E2F1-expressing adenovirus and examined the expression of more than 6000 genes using high-density microarrays. Microarray results clearly support the current paradigm; however, they suggest that E2F1 may also regulate unanticipated cellular functions including pathways involved in apoptosis, signal transduction, transcriptional control, and membrane biology. Most surprisingly, we identified a number of genes that are repressed by E2F1 expression, suggesting that E2F1 may have the potential to repress transcription of numerous genes through an unknown mechanism. PMID- 11888209 TI - Purification and characterization of isocitrate lyase from the wood-destroying basidiomycete Fomitopsis palustris grown on glucose. AB - Isocitrate lyase (EC 4.1.3.1), a key enzyme in the glyoxylate cycle, was purified 76-fold with 23% yield as an electrophoretically homogeneous protein from the wood-destroying basidiomycete Fomitopsis palustris grown on glucose. The native enzyme has a molecular mass of 186 kDa, consisting of three identical subunits of 60 kDa. The K(m) for DL-isocitrate was found to be 1.6 mM at the optimum pH (7.0). The enzyme required Mg(2+) (K(m) 92 microM) and sulfhydryl compounds for optimal activity. The enzyme activity was strongly inhibited by oxalate and itaconate with a K(i) of 37 and 68 microM, respectively. The inhibition by the glycolysis and tricarboxylic acid cycle intermediates and related compounds suggested that the isocitrate lyase was a regulatory enzyme playing a crucial role in the fungal growth. PMID- 11888210 TI - Cloning and characterization of an S-formylglutathione hydrolase from Arabidopsis thaliana. AB - A cDNA from Arabidopsis thaliana resembling S-formylglutathione hydrolase (SFGH), an enzyme with putative roles in formaldehyde detoxification in animals and microorganisms, has been cloned and expressed in Escherichia coli. The purified recombinant Arabidopsis enzyme (AtSFGH) was a dimer composed of 31-kDa subunits. Like SFGHs from other sources, AtSFGH had thioesterase activity toward S formylglutathione and carboxyesterase activity toward 4-methylumbelliferyl acetate. Unlike other SFGHs, the enzyme from Arabidopsis actively hydrolyzed S acetylglutathione. AtSFGH activity was inhibited by heavy metals and sulfhydryl alkylating agents, but was insensitive to serine hydrolase inhibitors, suggesting that the enzyme was a cysteine-dependent hydrolase. Although Atsfgh transcripts were determined in plants and cultures of Arabidopsis, the respective enzyme could not be detected in planta after the esterase activities present were resolved using isoelectric focusing. Instead, Arabidopsis contained several carboxyesterases active toward alpha-naphthyl acetate, which were all sensitive to inhibition by the serine hydrolase inhibitor paraoxon. PMID- 11888211 TI - Calcium-mediated telomerase activity in ovarian epithelial cells. AB - Though the potential of telomerase as an anti-cancer target is evident, information about regulation of telomerase remains fragmentary. In the present study, we examined the role of calcium, an essential cellular signaling molecule, in the regulation of telomerase. We found that calcium induced de novo telomerase activity in telomerase-negative ovarian surface epithelial (OSE) cell lines but not in primary cultures of OSE. In addition, we showed that calcium elevated endogenous telomerase levels in a telomerase-positive ovarian cancer cell line. The use of calcium channel blockers or calcium chelators inhibited this calcium mediated induction of telomerase activity. Furthermore, cadmium and chromium appeared to cause a moderate induction of telomerase activity while several other metal salts did not. Our data provide the first example of calcium-induced telomerase activity in human cell lines, provide a novel avenue for possible intervention of telomerase, and permit development of therapeutic agents for adjunctive chemotherapy. PMID- 11888212 TI - Cellular adhesion of primary Sertoli cells affects responsiveness of the extracellular signal-regulated kinases 1 and 2 to follicle-stimulating hormone but not to epidermal growth factor. AB - The cellular adhesion status and the exposure to soluble growth factors both contribute to mitogen-activated protein (MAP) kinase activation. To date, however, whether mitogens acting through G-protein-coupled receptors (GPCRs) need cell adhesion to activate MAP kinases/extracellular signal-regulated kinases (ERK) 1, 2 has been poorly investigated. We addressed this point in primary cultures of Sertoli cells experimentally maintained in suspension, considering that follicle-stimulating hormone (FSH) activates ERK1, 2 in attached Sertoli cells by acting through a GPCR. We found that FSH actively repressed ERK1, 2, in a cAMP-dependent but cAMP-dependent protein kinase (PKA)-independent manner, and this inhibition required the activity of a tyrosine phosphatase. In comparison, in the absence of anchorage, ERK1, 2 were still activated by epidermal growth factor, in a PKA-dependent manner. Altogether, these data suggest that sensitivity of the MAP kinase response toward cell adhesion may depend, at least in part, on the class of receptor, GPCR or receptor with tyrosine kinase activity, by which it is triggered. PMID- 11888213 TI - Operating conditions that affect the resistance of lactic acid bacteria to freezing and frozen storage. AB - Thermophilic lactic acid bacteria exhibit different survival rates during freezing and frozen storage, depending on the processing conditions. We used a Plackett and Burman experimental design to study the effects of 13 experimental factors, at two levels, on the resistance of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to freezing and frozen storage. The resistance was evaluated by quantifying the decrease of acidification activity during freezing and throughout 8 weeks of storage. Acidification activity after freezing and frozen storage was affected by 12 experimental factors. Only the thawing temperature did not show any significant effect. S. thermophilus was more resistant than L. bulgaricus and the cryoprotective effect of glycerol during freezing and storage was confirmed. The temperature and duration of the cryoprotection step influenced acidification activity following the freezing step: the lower the temperature and the shorter the duration, the higher the activity. Acidification activity after storage was affected by several experimental factors involved in the fermentation stage: use of NaOH instead of NH4OH for pH control, addition of Tween 80 in the culture medium, and faster cooling led to better cryotolerance. Resistance to freezing and frozen storage was improved by using a high freezing rate and a low storage temperature. Finally, this study revealed that the conditions under which lactic acid bacteria are prepared should be well controlled to improve their preservation and to limit the variability between batches and between species. PMID- 11888214 TI - Effect of salts on the properties of aqueous sugar systems, in relation to biomaterial stabilization. 1. Water sorption behavior and ice crystallization/melting. AB - Trehalose and sucrose, two sugars that are involved in the protection of living organisms under extreme conditions, and their mixtures with salts were employed to prepare supercooled or freeze-dried glassy systems. The objective of the present work was to explore the effects of different salts on water sorption, glass transition temperature (T(g)), and formation and melting of ice in aqueous sugar systems. In the sugar-salt mixtures, water adsorption was higher than expected on the basis of the water uptake by each pure component. In systems with a reduced mass fraction of water (w less-than-or-equal 0.4), salts delayed water crystallization, probably due to ion-water interactions. In systems where > 0.6, water crystallization could be explained by the known colligative properties of the solutes. The glass transition temperature of the maximally concentrated matrix (T(g)') was decreased by the presence of salts. However, the actual T(g) values of the systems were not modified. Thus, the effect of salts on sorption behavior and formation of ice may reflect dynamic water-salt-sugar interactions which take place at a molecular level and are related to the charge/mass ratio of the cation present without affecting supramolecular or macroscopic properties. PMID- 11888215 TI - Laboratory studies of cryopreservation of sperm and trochophore larvae of the eastern oyster. AB - The eastern oyster, Crassostrea virginica, is the most important cultured oyster species of the Atlantic and Gulf coasts of the United States. Cryopreservation of gametes and larvae of aquatic organisms has increased in importance in recent years. However, studies on the cryopreservation of sperm and larvae of mollusks have focused on the Pacific oyster, Crassostrea gigas. The present study was conducted to improve cryopreservation of sperm and trochophore larvae and to assess fertilizing ability and male-to-male variation of thawed sperm of the eastern oyster. Sperm were diluted in 12 cryoprotectant solutions composed of Hanks' balanced salt solution without calcium and 0, 5, 10, 15, 20, and 25% (v/v) propylene glycol with or without 0.25 M sucrose. Trochophore larvae were suspended in artificial seawater and 10 or 15% propylene glycol (v/v). Sperm or trochophore larvae were placed in 5-mL macrotubes and allowed to equilibrate for 15 min. The macrotubes were cooled in a controlled-rate freezer at a rate of 2.5 degrees C per min until reaching a final temperature of -30 degrees C and were plunged into liquid nitrogen. After storage for 2 weeks, the samples were thawed in a water bath at 70 degrees C for 15 s. Overall, for cryopreservation of sperm and larvae, best results were obtained using 10 or 15% propylene glycol. Thawed sperm presented significant male-to-male variation in fertilizing ability. Survival of thawed larvae decreased as the concentration of larvae per macrotube increased. The procedures developed in this study for sperm and larvae are suitable for production of seedstock in commercial oyster hatcheries. PMID- 11888216 TI - Water crystallization within rat precision-cut liver slices in relation to their viability. AB - This study examined whether tissue vitrification, promoted by partitioning within the tissue, could be the mechanism explaining the high viability of rat liver slices, rapidly frozen after preincubation with 18% Me2SO or VS4 (a 7.5 M mixture of Me2SO, 1,2-propanediol, and formamide with weight ratio 21.5:15:2.4). To achieve this, we first determined the extent to which crystallization or vitrification occurred in cryoprotectant solutions (Me2SO and VS4) and within liver slices impregnated with these solutions. Second, we determined how these events were related to survival of slices after thawing. Water crystallization was evaluated by differential scanning calorimetry and viability was determined by histomorphological examination of the slices after culturing at 37 degrees C for 4 h. VS4-preincubated liver slices indeed behaved differently from bulk VS4 solution, because, when vitrified, they had a lower tendency to devitrify. Vitrified VS4-preincubated slices that were warmed sufficiently rapid to prevent devitrification had a high viability. When VS4 was diluted (to 75%) or if warming was not fast enough to prevent ice formation, slices had a low viability. With 45% Me2SO, low viability of cryopreserved slices was caused by cryoprotectant toxicity. Surprisingly, liver slices preincubated with 18% Me2SO or 50% VS4 had a high viability despite the formation of ice within the slice. In conclusion, tissue vitrification provides a mechanism that explains the high viability of VS4 preincubated slices after ultrarapid freezing and thawing (>800 degrees C/min). Slices that are preincubated with moderately concentrated cryoprotectant solutions (18% Me2SO, 50% VS4) and cooled rapidly (100 degrees C/min) survive cryopreservation despite the formation of ice crystals within the slice. PMID- 11888217 TI - A comparison of the metabolic effects of continuous hypothermic perfusion or oxygenated persufflation during hypothermic storage of rat liver. AB - The metabolic consequences of supplying oxygen by two different modes were investigated. The effects of hypothermic liver preservation after cold hypoxic flush (Group I), oxygenated vascular persufflation (Group II), and continuous oxygenated perfusion (Group III) were compared. Adenine nucleotides were measured to assess energetics, and 1H nuclear magnetic resonance spectroscopy was employed to investigate other metabolic pathways. Energetics were maintained by both modes of oxygenation at 24 h. The mitochondrial redox state is indicated by the ratio of acetoacetate (Ace) and beta-hydroxybutyrate (betaHb). The detection of only betaHb or Ace in the hypoxic flush and perfused livers, respectively, suggested that the mitochondria of these livers were hyperreduced and hyperoxidized, respectively. In contrast, both components of the redox couple were detected in the persufflated livers, suggesting that persufflation may be a simple and effective method of maintaining hepatic energetics long-term while maintaining a more normal mitochondrial redox state. PMID- 11888218 TI - Starfish oocytes form intracellular ice at unusually high temperatures. AB - Starfish oocytes, eggs, and embryos are popular models for studying meiotic maturation, fertilization, and embryonic development. Their large (170- to 200 microm) oocytes are obtainable in copious amounts and are amenable to manipulations that mammalian oocytes are not. The most formidable obstacle to working with marine oocytes is their seasonal availability, yet a successful means of preserving them for use during the nonreproductive season has not been reported. The aim of this study was to investigate the response of starfish oocytes to freezing with rapid and slow cooling rates under a variety of conditions to develop a cryopreservation protocol for these cells. Cryomicroscopic observation revealed that starfish oocytes in isotonic medium undergo intracellular ice formation (IIF) at very high subzero temperatures, such that the mean difference between the temperature of extracellular ice formation (T(EIF)) and IIF (TI(IF)) was less than 3 degrees C and the average T(IIF) was approximately between -4 and -6 degrees C. Neither partial cellular dehydration nor addition of the cryopreservative dimethyl sulfoxide significantly depressed the T(IIF). Under some conditions, we observed ice nucleation at multiple locations within the cytoplasm, suggesting that several factors contribute to the unusually high T(IIF) during controlled-rate freezing and thus vitrification may be a more suitable method for cryopreserving these cells. PMID- 11888219 TI - Transplantation of articular cartilage following a step-cooling cryopreservation protocol. AB - Using a step-cooling cryopreservation protocol that held the tissue 60 min at -4 degrees C, 30 min at -8 degrees C, and 10 min at -40 degrees C before plunging into liquid nitrogen, we were able to get a substantial improvement in the magnitude and pattern of chondrocyte recovery following cryopreservation, achieving postthaw recoveries of 62 +/- 13%. These results are consistent with the hypothesis that ice growth within articular cartilage is planar, but they provide no direct support for that hypothesis. Transplanting (step-cooled) cryopreserved osteochondral allografts into adult Suffolk/Romanoff crossbred sheep for periods of 3 months and 1 year further tested the efficacy of the cryopreservation protocol. Unfortunately, the cryoinjury sustained by the chondrocytes during cryopreservation, although apparently nonlethal immediately after thawing in many cases, was not innocuous in the long term. The presence of large clusters of chondrocytes at 1 year after transplantation illustrates that cryoinjury not detectable with a membrane integrity assay can still have far reaching effects on transplanted tissue. PMID- 11888220 TI - Magnetic resonance imaging-estimated three-dimensional temperature distribution in liver cryolesions: a study of cryolesion characteristics assumed necessary for tumor ablation. AB - The goal of this study was to estimate the three-dimensional (3D) temperature distribution in liver cryolesions and assess the margin of the transition zone between the tumoricidal core of the lesion and the surrounding unfrozen tissue, using criteria proposed in the literature. Local recurrences after liver tumor cryoablation are frequent. Temperatures below -40 degrees C and a 1-cm zone of normal tissue included in the cryolesion are considered necessary for adequate ablation. The 3D temperature distribution in 10 pig cryolesions was estimated by numerical solution of a simplified bioheat equation using magnetic resonance imaging data to establish cryolesion border conditions. Volumes encompassed by the -20, -40, and -60 degrees C isotherms were estimated. The shortest distance from every voxel on the -40 degrees C isotherm to the cryolesion edge was calculated and the mean and the maximal of these distances were defined for each cryolesion. Median cryolesion volumes with temperatures of -20, -40, and -60 degrees C or colder were 53, 26, and 14% of the total cryolesion volume, respectively. The median cryolesion volume was 12.3 cm(3). The median of the mean distances calculated between the -40 degrees C isotherm and the cryolesion edge was 4.1 mm and increased with increasing cryolesion volume. The median of the largest of these distances calculated for each cryolesion was 8.1 mm. Temperatures claimed to be adequate for tumor destruction were obtained only in parts of the cryolesion. The adequacy of a 1-cm zone of normal liver tissue included in the cryolesion to ensure tumor ablation is questioned. PMID- 11888221 TI - Influence of cooling rates and plunging temperatures in an interrupted slow freezing procedure for semen of the African catfish, Clarias gariepinus. AB - The objective of this study was to optimize interrupted slow-freezing protocols for African catfish semen. Semen diluted with methanol and extender was frozen in 1-ml vials in a programmable freezer. The temperatures of the freezer (T(chamber)) and of the semen (T(semen)) were measured simultaneously. We first tested two-step freezing protocols with different cooling rates (-2, -5, and -10 degrees C/min) and different temperatures at plunging into liquid N2. The difference between T(semen) and T(chamber) increased with faster cooling rates. In all programs, survival of spermatozoa, expressed as hatching rates, increased from near zero when T(semen) at plunging was higher than -30 degrees C to values equal to those of control when T(semen) at plunging was equal to or lower than 38 degrees C. The inclusion of an isothermal holding period before plunging into liquid N2 (three-step freezing protocols) resulted in an equilibration between T(semen) and T(chamber) and improved semen survival. Semen could be plunged at temperatures as high as -36 degrees C when cooled at -5 or -10 degrees C/min, without compromising postthaw semen survival. Cooling at -2 degrees C/min in combination with a 5-min holding period reduced postthaw survival. We conclude that with slow cooling rates of -2 to -5 degrees C/min, hatching rates can be maximized by plunging as soon as T(semen) reaches -38 degrees C. The isothermal holding period is beneficial when faster rates are used. A simple and efficient protocol for freezing African catfish semen can be obtained by cooling at a rate of -5 to -10 degrees C/min combined with a 5-min holding period in the freezer, at -40 degrees C. PMID- 11888222 TI - Exposure to dimethyl sulfoxide at 37 degrees C prior to freezing significantly improves the recovery of cryopreserved hybridoma cells. AB - Standard cryopreservation protocols recommend the use of dimethyl sulfoxide (Me2SO) at moderate temperatures only (room temperature, 4 degrees C) due to its toxicity which appears to be potentiated by warm temperatures. In the present study, we asked whether a transient increase in temperature during membrane sealing of cryovials affects the cell viability. We show here that the cell viability of hybridoma cells and Schwann cells was not reduced following membrane sealing of cryotubes. On the contrary, incubation of cells at 37 degrees C in Me2SO-containing medium prior to freezing significantly stimulated the viability of cryopreserved hybridoma cells, whereas the viability of Schwann cells remained unaltered. We conclude that the exposure of cells to Me2SO at elevated temperatures does not necessarily reduce cell viability and that contrary to this, cell type-specific, beneficial effects of Me2SO could be observed. PMID- 11888223 TI - Differential effectiveness of psychological interventions for reducing osteoarthritis pain: a comparison of Erikson [correction of Erickson] hypnosis and Jacobson relaxation. AB - The present study investigates the effectiveness of Erikson hypnosis and Jacobson relaxation for the reduction of osteoarthritis pain. Participants reporting pain from hip or knee osteoarthritis were randomly assigned to one of the following conditions: (a) hypnosis (i.e. standardized eight-session hypnosis treatment); (b) relaxation (i.e. standardized eight sessions of Jacobson's relaxation treatment); (c) control (i.e. waiting list). Overall, results show that the two experimental groups had a lower level of subjective pain than the control group and that the level of subjective pain decreased with time. An interaction effect between group treatment and time measurement was also observed in which beneficial effects of treatment appeared more rapidly for the hypnosis group. Results also show that hypnosis and relaxation are effective in reducing the amount of analgesic medication taken by participants. Finally, the present results suggest that individual differences in imagery moderate the effect of the psychological treatment at the 6 month follow-up but not at previous times of measurement (i.e. after 4 weeks of treatment, after 8 weeks of treatment and at the 3 month follow-up). The results are interpreted in terms of psychological processes underlying hypnosis, and their implications for the psychological treatment of pain are discussed. PMID- 11888224 TI - Venlafaxine in neuropathic pain following treatment of breast cancer. AB - Amitriptyline effectively relieves neuropathic pain following treatment of breast cancer. However, adverse effects are a major problem. Venlafaxine has no anticholinergic effects and could have a better compliance. The aim of the study was to evaluate the effectiveness of venlafaxine in neuropathic pain. The study was a randomized, double-blind, crossover comparison of venlafaxine and inactive placebo. The study lasted 10 weeks. The number of tablets (18.75 mg) taken daily was increased by one at a 1 week interval. Pain intensity and pain relief were registered daily by a diary and by a questionnaire and a computer program (Painscreen) on each visit. Adverse effects were evaluated with the diaries and a 10-item list on each visit. Also, anxiety and depression were measured on each visit. Venous blood samples were collected before the treatment and at 4 weeks for the determination of the serum levels of venlafaxine and its three metabolites. Thirteen patients were analysed. The average daily pain intensity as reported in the diary (primary outcome) was not significantly reduced by venlafaxine compared with placebo. However, the average pain relief (diary) and the maximum pain intensity (retrospective assessment by the computer program) were significantly lower with venlafaxine compared with placebo. Anxiety and depression were not affected. Adverse effects did not show significant differences between treatments. The two poor responders had low venlafaxine concentrations whereas the two slow hydroxylizers had high venlafaxine concentrations and excellent pain relief. Thus, higher doses could be used in order to improve pain relief. PMID- 11888226 TI - Clonidine for treatment of postoperative pain: a dose-finding study. AB - The aim of this double-blind randomized study was to evaluate the optimal intravenous dose of clonidine administrated during the peri-operative period, after lumbar hemilaminectomy for herniated disk repair. The "optimal intravenous dose" was defined as that providing minimal analgesic request, stable haemodynamic profile and a minimal sedation score during 12h after extubation. Eighty adult patients, ASA physical status I-II, undergoing lumbar hemilaminectomy for herniated disk (L(4)-L(5), L(5)-S(1)) were included in the study. All the patients were randomly assigned to one of four study groups (A, B, C, D), 20 patients each. The same standardized general anaesthesia was performed for each group. Thirty minutes before the end of surgery, group A, B and C patients received three different loading doses of intravenous clonidine (5 microg/kg, 3 microg/kg, 2 microg/kg respectively), followed by the same infusion of intravenous clonidine (0.3 microg/kg per hour). Group D patients received a bolus dose and a continuous infusion of NaCl 0.9%. In the recovery unit, postoperative pain was treated by a patient-controlled analgesia device, containing morphine. Pain relief was evaluated by the total morphine requirement during the postoperative period. Systolic blood pressure (SBP), heart rate and sedation were also noted during the first 12h postoperatively. Intravenous clonidine decreased morphine requirements in a dose-dependent manner. Group A, B, C and D patients requested 5 +/- 2, 11 +/- 3, 19 +/- 4 and 29 +/- 8 doses of morphine respectively. Clonidine also affected SBP in a dose-related manner. Group A, B and C patients had an SBP decrease respectively of 26 +/- 3%, 7 +/- 4% and 2 +/- 2% compared with basic values while, at the same time, in group D patients no SBP variation was registered. In conclusion, this study demonstrates that, when sedation and analgesic effect of clonidine is required, 3 microg/kg bolus dose followed by a continuous infusion of 0.3 microg/kg per hour has to be considered the optimal intravenous dose. The higher dose of intravenous clonidine (5 microg/kg) produced better analgesia but the degree of hypotension and sedation was more severe and longer lasting; it required ephedrine administration and careful monitoring of the patient. On the other hand, the bolus of intravenous clonidine 2 microg/kg (group C) was less effective in terms of pain relief but with similar side-effects to the 3 microg/kg dosage (group B). PMID- 11888225 TI - Subcutaneous formalin and intraplantar carrageenan increase nitric oxide release as measured by in vivo voltammetry in the spinal cord. AB - The paper describes in vivo voltammetric detection of nitric oxide with carbon fibre microelectrodes at the lumbar spinal dorsal horn level of decerebrated spinalized rats during peripheral noxious inflammatory processes. At the lumbar (L3-L4) dorsal horn level, a nitric oxide dependent peak of oxidation current (650 mV), remaining stable for up to 4h ((92 +/- 5)% of control) could be detected indicating that significant amounts of nitric oxide are produced continuously. Following subcutaneous injection in the hindpaw of 50 microl of 0.5% formalin the oxidation current rapidly increased ((115 +/- 5)% of control at 25 min) and reached (120 +/- 6)% of control 1h later. Subsequently the voltammograms stabilized for up to 90 min and decreased ((107 +/- 4)% at 124 min). After an injection in the hindpaw of 150 microl of 4% carrageenan, the voltammograms remained at control level for 1h and then the oxidation current increased continuously for up to 4h ((145 +/- 16)% of control at 240 min); such an increase was reversed by ketamine. In these two models of inflammation, the delay in onset and the duration of the increases in NO release within the dorsal horn relate, to some extent, to the time course of the peripheral inflammatory processes, since they are shorter after formalin than after carrageenan. The results provide a direct in vivo demonstration that the intercellular messenger nitric oxide participates in the transmission of noxious afferent messages within the dorsal horn of the spinal cord following peripheral inflammation. PMID- 11888227 TI - Preliminary validation of a German version of pain stages of change questionnaire. AB - This study describes the development of the German questionnaire FF-STABS (Freiburg Questionnaire--Stages of Chronic Pain Management), which documents the willingness of chronic pain patients to use cognitive-behavioural methods for pain management independently. The newly constructed assessment instrument, modified from a similar instrument developed by Kerns and his colleagues, was administered to a heterogeneous sample of 118 chronic pain patients. Item and factor analysis support the identification of four reliable scales: precontemplation, preparation, action and maintenance. All scales evidenced sufficient indices of reliability and discriminant validity. Deviations from the original American version and suggestions for future refinements in the new measure are discussed. PMID- 11888228 TI - Calculation methods for the continuous intrathecal infusion of a solution containing two or three drugs. AB - The continuous intrathecal infusion of drugs with implantable and programmable pumps is commonly used for the treatment of otherwise intractable pain and spasticity. There is now a consensus that it is possible to use mixtures of two or even three drugs in selected cases. The calculation of the parameters of the pump is easy for a single drug, but it becomes more complicated for a combination of two or three drugs. This paper offers some algebraic formulae that make easier and faster the calculation of the following values: the flow of the pump, the volume of each drug solution to be added to the final mixture, the concentration of each drug in the mixture and the duration of the infusion. These values are determined from the desired quantity of each drug to be infused in 24h, the concentration of each drug solution and the volume of the reservoir. It is essential that the drugs do not react with one another, that they will remain stable in the reservoir at body temperature and that they are safe for a long term infusion. PMID- 11888229 TI - Nociceptive behaviour induced by dental application of irritants to rat incisors: a new model for tooth inflammatory pain. AB - Animal models simulating acute human pulpitis are still lacking. The rat incisors present a particular situation where most of their innervation is considered to be unmyelinated and concentrated mainly in the tooth pulp. This study reports on a new model for dental pain induced by inflammatory agents applied to the tooth pulps of incisors. In different groups of rats, artificial crowns were fixed on the lower incisors, after cutting 1-2mm of their distal extremities. A volume of 7-10 microl of solutions of saline, capsaicin (1-10mg/ml) or formalin (2.5% or 5%) was injected in the crown cavity, and the nociceptive behaviour was quantitated following a devised scoring method of four scales. Intradental application of capsaicin produced nociceptive scores in the form of one plateau for 1-2h depending on the concentration used. Similar results were obtained with intradental application of formalin 2.5%. The one plateau of nociceptive scores obtained with formalin contrasts with the biphasic aspect of nociceptive behaviour described with the intradermal formalin test. This discrepancy could be attributed to a difference in the types of afferent fibres involved in each situation. Pretreatment with morphine (2 mg/kg) attenuated, in a naloxone reversible manner, the nociceptive behaviour observed following intradental application of capsaicin. Pretreatment with meloxicam (a cyclo-oxygenase-2 inhibitor) exerted a less pronounced attenuation of the nociceptive scores when compared with morphine. These results provide evidence for the validity of the described model for the simulation of tooth pulp inflammatory pain in awake animals. PMID- 11888230 TI - Morphine responsiveness in a group of well-defined multiple sclerosis patients: a study with i.v. morphine. AB - Pain in multiple sclerosis (MS) is more common than has previously been believed. About 28% of all MS patients suffer from central pain (CP), a pain that is difficult to treat. In the present study we have investigated the responsiveness of this pain to morphine. Fourteen opioid-free patients (eight woman and six men) with constant, non-fluctuating, long-lasting CP caused by MS were investigated. Placebo (normal saline), morphine and naloxone were given intravenously in a standardized manner. The study design was non-randomized, single blind and placebo controlled. Ten patients experienced less than 50% pain reduction by placebo and less than 50% pain reduction by morphine. Four patients were opioid responders, i.e. had minimal or no effect on pain by placebo, >50% pain reduction after morphine and >25% pain increase after naloxone, given intravenously following morphine. However, this response was obtained after high doses of morphine (43 mg, 47 mg, 50 mg and 25 mg; mean 41 mg). Thus, compared with nociceptive pain, only a minority of the patients with CP due to MS responded to morphine and only at high doses. The present results are in accord with experimental studies indicating that neuropathic pain is poorly responsive but not totally unresponsive to opioids. The results do not support the routine use of strong opioids in MS patients with CP. PMID- 11888232 TI - Five easy pieces on evidence-based medicine (4). PMID- 11888231 TI - Systemic morphine selectively depresses a thalamic link of widespread nociceptive inputs in the rat. AB - The lateral part of the ventromedial thalamus (VM l) relays nociceptive inputs from the whole body surface to the dorsolateral frontal cortex. The aim of the present study was to investigate the effects of systemic morphine on nociceptive activity evoked in VM l neurones either by thermal (48 degrees C) or by supramaximal percutaneous electrical stimuli. The noxious thermal evoked responses were depressed by 10.8 +/- 10.1%, 48.3 +/- 23.0% and 67.3 +/- 10.1%, 5 min after i.v. injections of 1.0, 1.73 and 3.0 mg/kg of morphine, respectively. Moreover, strong depressive effects on the Adelta- and C-fibre responses were already present 5 min after the injection. The responses were significantly reduced by 7.2 +/- 5.9%, 32.5 +/ 11.1% and 37.2 +/- 11.8% for Adelta fibres after i.v. injections of 1.0, 1.73 and 3.0 mg/kg of morphine, respectively. The corresponding values for C-fibre evoked responses were 16.3 +/- 16.2%, 57.0 +/- 12.0% and 69.0 +/- 8.2%. The dose of morphine that reduced VM l neuronal nociceptive responses by 50% (1.73 mg/kg) was around 3.5 times lower than that necessary to inhibit the responses of its spinal or medullary relays under similar experimental conditions. These results, added to the data of the literature, suggest that supraspinal effects of morphine are primarily mediated at the thalamic level. It is tempting to speculate that morphine-induced reductions of attentional or psychomotor responses related to pain may be mediated by its action on VM l. PMID- 11888234 TI - Anticonvulsants: aspects of their mechanisms of action. AB - An ideal anticonvulsant drug would prevent or inhibit excessive pathological neuronal discharge without interfering with physiological neuronal activity and without producing untoward effects. Such an ideal compound is not yet available. However, during the last few years several new anticonvulsants have appeared (e.g. vigabatrine, gabapentin, topiramate, lamotrigine, tiagabine, felbamate and oxcarbazepine) which may challenge the older, more established substances (i.e. phenytoin, benzodiazepines, phenobarbital, valproate, carbamazepine and ethosuximide). Interestingly, several of the old and new anticonvulsants are beneficial in the treatment of various psychiatric conditions (most notably mood disorders) as well as neuropathic pain. The reason these various drugs are effective in the treatment of such disparate clinical conditions is unknown. The answer may be that the neuronal dysfunctions underlying these conditions are similar in a mechanistic sense, but are manifested in different neurons/locations of the nervous system, or that the drugs possess several mechanisms of action that contribute in different ways to the favourable effect depending on the condition studied. Even though all these drugs inhibit excessive neuronal activity, this acute effect appears to be produced by several mechanisms, which fall into three major categories: (1) blockade of voltage-gated sodium channels; (2) indirect or direct enhancement of inhibitory gamma-aminobutyric acid [GABAergic] neurotransmission; or (3) inhibition of excitatory glutamatergic neurotransmission. Moreover, several of these drugs fall into more than one category, and it is often unclear which category is responsible for a given effect of a drug. It is plausible that some of the beneficial effects observed in the clinic can be explained by the secondary neural depressant mechanisms of action of these substances, whereas other benefits may be due to long-term neuroplastic effects, which may either be common or different across the various conditions treated. PMID- 11888235 TI - Neuropathic pain: evidence matters. AB - "He slept less and less; they gave him opium and began to inject morphine. But this did not relieve him. The dull pain he experienced in the half asleep condition at first only relieved him as a change, but then it became as bad, or even more agonizing, than the open pain."--Tolstoy, The Death of Ivan Ilyitch. It comes as a surprise to many pharmacologists that not all pains are relieved by opioids. Those who work in chronic pain are unfortunately only too aware of the problems that such pains can cause. One of the hallmarks of neuropathic pain is poor or incomplete relief with opioids. As with so many things in medicine, there is nothing novel in this realization, as the Tolstoy quotation shows. PMID- 11888237 TI - Epilepsy: a clinical diagnostic overview. PMID- 11888238 TI - Channelopathies can cause epilepsy in man. AB - Idiopathic epilepsies, which account for up to 40% of all epilepsies, are mainly caused by genetic factors. Most idiopathic epilepsies are due to oligogenic or multifactorial rather than monogenetic inheritance. Nevertheless, most of what is known today about the molecular genetics of idiopathic epilepsies has been found by analysing large families with rare monogenetic forms of the disease. For the first time, gene defects can be linked to certain epilepsies. Mutations in the CHRNA4 or CHRNB subunits of the neuronal nicotinic acetylcholine receptor lead to familial nocturnal frontal lobe epilepsy, while defects in the voltage-gated potassium channels KCNQ2 and KCNQ3 have recently been found to cause benign familial neonatal convulsions. The voltage-gated sodium channel subunits SCN1B, SCN1A and SCN2A as well as the GABRG2 subunit of the GABA(A) receptor are involved in the pathology of the newly described syndrome generalized epilepsy with febrile seizures plus. These rare monogenetic epilepsies can serve as models for further genetic analysis of the common forms of idiopathic epilepsies. PMID- 11888239 TI - What can we learn from clinical trials of anticonvulsant drugs in epilepsy? AB - Any physician who intends to utilize the available antiepileptic drugs (AEDs) judiciously, cannot do so without being well versed on their pharmacological properties and the large body of evidence that is continuously accumulating on their relative efficacy and tolerability in different types of epilepsy. While informal observations such as retrospective surveys and case reports can be useful under special circumstances, prospective randomized clinical studies represent by far the most important tool by which objective information can be obtained about the clinical value of existing drugs. Even randomized trials, however, can produce misleading conclusions because of inherent weaknesses or bias in study design, analysis, and interpretation. Common deficiencies identified in some of the most recent drug trials in epilepsy include 1) inclusion of inappropriately heterogeneous patient groups (for example, patients with partial and primarily generalized seizures); 2) low statistical power due to insufficient sample size (for trials designed to show therapeutic equivalence); 3) inappropriate titration rates or suboptimal dosages or dosing schedules (often favouring the sponsor's product over the comparator); 4) insufficient duration of treatment; and 5) utilization of endpoints of questionable clinical significance. In part, some of the above shortcomings can be ascribed to the fact that most clinical drug trials are designed to address regulatory needs rather than to provide the type of information required for rational prescribing. Physicians need to be alerted about the importance of these issues, and they should make every possible effort to interpret critically the medical literature on which they rely to guide and support their therapeutic decisions. PMID- 11888241 TI - Neuropathic pain: clinical characteristics and diagnostic workup. AB - Neuropathic pain is part of the neurological disease spectrum and may be an expression of severe medical pathology. Painful neuropathies have multiple disguises and may to a certain extent be mimicked by non-neurological pain conditions. Painful neuropathic conditions express themselves with spontaneous and/or abnormal stimulus-evoked pain. The diagnosis of peripheral or central neuropathic pain should be made only when the history and signs are indicative of neuropathy in conjunction with neuroanatomically correlated pain distribution and sensory abnormalities within the area of pain. A future mechanism-based classification of pain has recently been suggested to facilitate the development of mechanism-tailored treatment strategies. This is a sound approach and should be pursued. It is mandatory, however, to retain the traditional organ-based diagnostic workup, which should precede further in-depth characterization of specific pain mechanisms. Extensive preparatory work is needed on how to link certain symptoms and signs to specific mechanisms, as elucidated from animal studies, before we can introduce mechanism-coupled treatment strategies. PMID- 11888242 TI - Neurobiology of neuropathic pain: mode of action of anticonvulsants. AB - Anticonvulsants are widely used for the treatment of neuropathic pain. Here we review the evidence for a number of peripheral and central changes after nerve injury that may provide a basis for the mechanisms of action of anticonvulsant therapies. The roles of sodium channels, calcium channels, and central glutamate mechanisms are emphasized as the main targets for anticonvulsant drugs in neuropathic pain states. The focus of this article is on anticonvulsants; however, opioids and antidepressants can also be effective in increasing inhibitions to control of pain in a manner similar to that of the enhancement of gamma-aminobutyric acid (GABA) function by antiepileptic drugs. A brief account of these approaches to neuropathic pain is also given. PMID- 11888243 TI - Anticonvulsants in neuropathic pain: rationale and clinical evidence. AB - Neuropathic pain, whether of peripheral or central origin, is characterized by a neuronal hyperexcitability in damaged areas of the nervous system. In peripheral neuropathic pain, damaged nerve endings exhibit abnormal spontaneous and increased evoked activity, partly due to an increased and novel expression of sodium channels. In central pain, although not explored in detail, the spontaneous pain and evoked allodynia are also best explained by a neuronal hyperexcitability. The peripheral hyperexcitability is due to a series of molecular changes at the level of the peripheral nociceptor, in dorsal root ganglia, in the dorsal horn of the spinal cord, and in the brain. These changes include abnormal expression of sodium channels, increased activity at glutamate receptor sites, changes in gamma-aminobutyric acid (GABA-ergic) inhibition, and an alteration of calcium influx into cells. The neuronal hyperexcitability and corresponding molecular changes in neuropathic pain have many features in common with the cellular changes in certain forms of epilepsy. This has led to the use of anticonvulsant drugs for the treatment of neuropathic pain. Carbamazepine and phenytoin were the first anticonvulsants to be used in controlled clinical trials. Studies have shown these agents to relieve painful diabetic neuropathy and paroxysmal attacks in trigeminal neuralgia. Subsequent studies have shown the anticonvulsant gabapentin to be effective in painful diabetic neuropathy, mixed neuropathies, and postherpetic neuralgia. Lamotrigine, a new anticonvulsant, is effective in trigeminal neuralgia, painful peripheral neuropathy, and post-stroke pain. Other anticonvulsants, both new and old, are currently undergoing controlled clinical testing. The most common adverse effects of anticonvulsants are sedation and cerebellar symptoms (nystagmus, tremor and incoordination). Less common side-effects include haematological changes and cardiac arrhythmia with phenytoin and carbamazepine. The introduction of a mechanism-based classification of neuropathic pain, together with new anticonvulsants with a more specific pharmacological action, may lead to more rational treatment for the individual patient with neuropathic pain. PMID- 11888244 TI - Trypanosoma evansi: cloning and expression in Spodoptera frugiperda [correction of fugiperda] insect cells of the diagnostic antigen RoTat1.2. AB - A complementary DNA encoding the variant surface glycoprotein (VSG) of Trypanosoma evansi Rode Trypanozoon antigenic type (RoTat)1.2, currently used for experimental serological diagnosis of T. evansi infection in livestock, was cloned as a recombinant plasmid and sequenced. A recombinant baculovirus containing the coding region of RoTat1.2 VSG was constructed to express the protein in Spodoptera frugiperda [corrected] insect cells. From this, sufficient quantities of the recombinant protein are being produced for empirical and wide scale objective assessment of the diagnostic potential of this antigen. The gene encoding the RoTat1.2 VSG was shown by PCR to be present in the genomes of many different cloned isolates of T. evansi, but not T. brucei, from geographically separate regions of Africa, Asia, and South America. With the recombinant RoTat1.2 at hand, it is now possible to investigate the extent to which epitopes on this VSG are conserved among different T. evansi isolates. PMID- 11888245 TI - Characterization of the receptor for insulin-like growth factor on Leishmania promastigotes. AB - Insulin-like growth factor (IGF)-I constitutively present in the skin is one of the first growth factors that Leishmania parasites encounter after transmission to the vertebrate host. We have previously shown that IGF-I is a potent growth promoting factor for Leishmania parasites. IGF-I binds specifically to a single site putative receptor at the parasite membrane, triggering a cascade of phosphorylation reactions. In the present article we characterize the receptor for IGF-I on Leishmania (Leishmania) mexicana promastigotes. The receptor is a monomeric glycoprotein with a molecular mass of 65 kDa and is antigenically related to the alpha chain of human type 1 IGF-I receptor. Upon IGF-I stimulation the receptor undergoes autophosphorylation on tyrosine residues with activation of its signaling pathway. Activation of the IGF-I receptor also leads to phosphorylation of an 185-kDa molecule that is homologous to the substrate of the insulin receptor present in human cells, the insulin receptor substrate 1 (IRS 1). PMID- 11888246 TI - Echinococcus granulosus: regulation of leukocyte growth by living protoscoleces from horses, sheep, and cattle. AB - To determine whether living hydatid tissue can, like hydatid fluid, regulate leukocyte growth, T-cell, B-cell, and macrophage lines were cocultured with protoscoleces of Echinococcus granulosus and their growth was compared with that of control cultures by thymidine uptake estimates and chemiluminescent assays of cell number. Protoscoleces supported mitosis of IL-1-deprived D10 T cells, but did not increase D10 count. The action of protoscoleces was affected by the species and organ of their origin and the length of time in culture. Unusually marked mitotic reaction, unaffected by parasite age and origin, was recorded in the B-cell line, BSM, also without commensurate count increase, indicating that induced mitosis resulted in cell loss. It is concluded that protoscoleces can induce mitosis in B and T cells of particular lineages and that this is a potential means of producing the pathological proliferation and depletion of B- and T-cell areas which characterize local reaction to hydatids. PMID- 11888247 TI - Occurrence of a diploid type and a new first intermediate host of a human lung fluke, Paragonimus westermani, in Korea. AB - The biology, chromosome number, and karyotype of a lung fluke, Paragonimus westermani (Kerbert, 1878) collected in Haenam, Haenam-gun Chollanam-do, Korea were analyzed. We compared the size of metacercariae from Haenam with those taken from a crayfish collected at Youngam, Youngam-gun, Chollanam-do, Korea. The mean length of P. westermani metacercariae from Haenam was 300.3 microm and that from Youngam was 362.0 microm. Adult worms were recovered from the lungs of experimentally infected dogs. The mean egg sizes obtained from adult flukes were 72.1 x 46.8 microm from Haenam and 93.5 x 54.2 microm from Youngam. Semisulcospira tegulata collected in the Youngam area were found to be infected with cercariae of P. westermani, one of the snail-borne human lung fluke trematodes in Korea. Of 4218 snails studied, 5 (0.12%) harbored P. westrermani larvae. This is the first report of S. tegulata serving as the initial intermediate host of P. westermani. The chromosome numbers of P. westermani from Haenam and Youngam were 2n = 22 and 3n = 33. The diploid type of P. westermani has not been previously reported in Korea. PMID- 11888248 TI - Trypanosoma evansi: a convenient model for studying intracellular Ca(2+) homeostasis using fluorometric ratio imaging from single parasites. AB - The aim of this work was to measure, for the first time, the basal cytosolic Ca(2+) levels of Trypanosoma evansi and to explore the possibility of observing changes in the intracellular Ca(2+) concentration ([Ca(2+)](i)) using fluorescence ratio imaging techniques in single isolated parasites of this species. Under appropriate loading conditions, the high intracellular levels of the Ca(2+) fluorescence probe Fura-2 permits resolution, in real time, of single parasite [Ca(2+)](i) signals. Measurements of the basal [Ca(2+)](i) indicate that homeostatic mechanisms maintain [Ca(2+)](i) at 106 +/- 38 (n = 32) nM in the presence of 2 mM extracellular calcium. The resting [Ca(2+)](i) was unaffected by changes in extracellular Ca(2+) in the range from 0 to 10 mM. The Ca(2+) ionophore A23187 induced a large increase in [Ca(2+)](i) which (i) reached a steady state value even in the simultaneous presence of both external calcium and ionophore and (ii) returned to base line upon removal of extracellular Ca(2+). A dose-response curve of the protonophore nigericin shows that T. evansi contains an important pH-sensitive intracellular pool which may be released by this drug with a K(1/2) of 8 microM. These data demonstrate that this parasite contains highly efficient systems to control [Ca(2+)](i). Finally, our results, with the use of sera as source of an antibody-complement to induce Ca(2+) entry, demonstrate that it is possible to resolve fast [Ca(2+)](i) signals in single parasites from T. evansi. PMID- 11888249 TI - Leishmania donovani: expression and characterization of Escherichia coli expressed recombinant chitinase LdCHT1. AB - Leishmania parasites produce chitinase activity (EC. 3.2.1.14) thought to be important in parasite-sandfly interactions and transmission of the parasite to the vertebrate host. Previous observations have suggested that parasite chitinases are involved in degradation of the sandfly peritrophic matrix and the chitinous layer of the cardiac valve cuticle. This chitinase activity is thought to produce an incompetent pharyngeal valve sphincter and a route of egress that allow Leishmania promastigotes to be regurgitated into the site of blood feeding. In the studies reported here, enzymatically active L. donovani chitinase LdCHT1 was expressed as a thioredoxin fusion protein in Escherichia coli strain AD494 (DE3). Recombinant LdCHT1 had a predominantly endochitinase activity, in contrast to previous reports of both exo- and endochitinase activities in axenic culture supernatants of diverse Leishmania spp. promastigotes. The predominant endochitinase activity of recombinant LdCHT1 is consistent with the presumed function of the enzyme in disrupting chitinous structures in the sandfly digestive system to allow transmission. PMID- 11888250 TI - Entamoeba dispar: molecular characterization of the galactose/N-acetyl-d galactosamine lectin. AB - Amebiasis contributes to approximately 50 million cases of life-threatening dysentery worldwide. Comparison of the lectins from Entamoeba histolytica (pathogenic) and Entamoeba dispar (nonpathogenic) was undertaken to elucidate the differential roles of this molecule in invasion versus colonization. Surface lectin was less abundant on axenic E. dispar than on axenic E. histolytica, commensurate with differences in lectin (heavy and light subunits) RNA when assessed by semiquantitative RT-PCR. The 1G7 epitope, which falls within the immunodominant and immunoprotective cysteine-rich region (480-900), was absent on axenic E. dispar. Indirect immunofluorescence, transient transection of COS7, and immunoprecipitation demonstrated that the 1G7 epitope was conserved in the nonpathogenic lectin homologue but not exposed on live E. dispar trophozoites. Hgl2 (E. histolytica) and Dhgl2 (E. dispar) lectin homologues demonstrated comparable high-affinity binding to multivalent GalNAc(19) BSA. These data provide evidence for relative gene and conformational regulation of the E.dispar lectin. PMID- 11888251 TI - Toxoplasma gondii: mechanism of the parasitostatic action of 6-thioxanthine. AB - In contrast to the cytocidal effect of 6-thiopurines on mammalian cells, the action of 6-thioxanthine on Toxoplasma gondii was only parasitostatic. 6 Thioxanthine was a substrate of the parasite's hypoxanthine-guanine phosphoribosyltransferase. That enzyme converted 6-thioxanthine to 6 thioxanthosine 5'-phosphate which accumulated to near millimolar concentrations within parasites incubated intracellularly in medium containing the drug. 6 Thioxanthosine 5'-phosphate was the only detectable metabolite of 6-thioxanthine. The absence of 6-thioguanine nucleotides explains the lack of a parasitocidal effect because the incorporation of 6-thiodeoxyguanosine triphosphate into DNA is the mechanism of the lethal effect of 6-thiopurines on mammalian cells. Extracellular parasites that had accumulated a high concentration of 6 thioxanthosine 5'-phosphate incorporated more labeled hypoxanthine or xanthine into their nucleotide pools than did control parasites. The basis for this increased nucleobase salvage remains unexplained. It was not due to up-regulation of hypoxanthine-guanine phosphoribosyltransferase and could not be explained by reduced use of labeled nucleotides for nucleic acid synthesis. Extracellular parasites that had accumulated a high concentration of 6-thioxanthosine 5' phosphate used labeled hypoxanthine almost entirely to make adenine nucleotides while control parasites made both adenine and guanine nucleotides. Both extracellular parasites that had accumulated a high concentration of 6 thioxanthosine 5'-phosphate and control parasites efficiently used labeled xanthine to make guanine nucleotides. These observations suggested that inosine 5'-phosphate-dehydrogenase was inhibited while guanosine 5'-phosphate synthase was not. Assay of inosine 5'-phosphate dehydrogenase in soluble extracts of T. gondii confirmed that 6-thioxanthosine 5'-phosphate was an inhibitor. We conclude that 6-thioxanthine blocks the growth of T. gondii by a depletion a guanine nucleotides. PMID- 11888253 TI - How can a nurse intervention help people with newly diagnosed epilepsy? A qualitative study (of patients' views). AB - The aim was to describe the patients' views of the challenges posed by a new diagnosis of epilepsy and their assessment of a nurse intervention. Neurologists in South-East England referred patients into the study. Following a trial of a nurse intervention a subgroup of patients were purposefully identified for in depth interviews. Transcriptions of tape-recorded interviews were analysed using qualitative methodology. We found that younger people with epilepsy seemed to experience more trouble with driving, jobs and managing their lives in the context of new epilepsy, while older people saw epilepsy as just another illness to cope with. Patients reported difficulty in remembering what their doctors told them which they attributed partly to lack of time available in the consultation. They valued the time, and the technique of probing with explanations used by the nurse. The nurse intervention was seen as useful in making sense of symptoms, tests, risk management, and driving regulations and in helping manage their medicine taking. We conclude that people with newly diagnosed epilepsy face different challenges, some of which are related to their age at diagnosis. Patients reported help from the nurse with understanding the diagnosis, tests, risk management and taking their medication. Follow-up is necessary to measure behavioural effects on self-management in the long run. PMID- 11888252 TI - Plasmodium falciparum: differential display analysis of gene expression during gametocytogenesis. AB - With the Plasmodium falciparum genome sequencing near completion, functional analysis of individual parasite genes has become the major task of the postgenomic era. Understanding the expression patterns of individual genes is the initial step toward this goal. In this report, we have examined gene expression during gametocytogenesis of the malaria parasite, P. falciparum, using a modified differential display (DD) method. The modifications of this method include adjusting the dNTP mix, using upstream primers with higher AT contents, and reducing the extension temperature of the polymerase chain reaction (PCR). With a combination of 16 arbitrary upstream primers and 3 one-base-anchored oligo(dT) primers, we have successfully cloned 80 unique cDNA tags from stage IV-V gametocytes. Further analysis by dot blots and semiquantitative reverse transcriptase-PCR showed that at least 49 cDNAs had induced or elevated levels of expression in gametocytes. These results indicate that this modified DD procedure is suitable for large-scale identification of developmentally regulated genes in the AT-rich Plasmodium genome. PMID- 11888254 TI - The effects of adjunctive topiramate therapy on seizure severity and health related quality of life in patients with refractory epilepsy---a Canadian study. AB - Although reduction in seizure frequency is the most common endpoint used to assess the antiepileptic efficacy, seizure frequency alone does not provide a complete picture of effectiveness, particularly in patients with refractory epilepsy. The aim of our study was to assess the effects of topiramate on seizure severity and health-related quality of life (HRQL), in addition to standard efficacy measures, in an open, multicentre, 6-month trial of patients with epilepsy uncontrolled on antiepileptic drugs other than topiramate. Two hundred and nine patients were enrolled and received topiramate for up to 6 months (initiated at 50 mg/day and titrated to a recommended dose of 200-400 mg/day) in addition to existing medication. The median reduction in seizure frequency from baseline to the post-titration period was 40.9% ( P< 0.0001). Patients also demonstrated a mean reduction in the Liverpool Seizure Severity Scale (LSSS) of 5.3 ( P< 0.0001), which was considered clinically significant. Statistically significant changes in HRQL were not observed with the SF-36, a generic measure. Tolerability of antiepileptic medication was good, with a low incidence of cognitive adverse events. The results indicate that topiramate significantly reduces seizure severity---an important aspect of HRQL---when administered as adjunctive therapy to anticonvulsant therapy. PMID- 11888255 TI - Idiopathic familial temporal lobe epilepsy with febrile convulsions. AB - Efforts to duplicate the genetic and molecular breakthroughs of autosomal dominant frontal lobe epilepsy have lead to increased interest in familial temporal lobe epilepsy. In this report we describe three kindreds. The epilepsy syndrome described manifests after the teenage years and was generally mild and treatment responsive. The predominant seizure types were simple and complex partial seizures, typical of mesial temporal onset. Some family members had febrile convulsions only and others had epilepsy without preceding febrile convulsions. Three patients had both febrile convulsions and temporal lobe epilepsy. High-resolution quantitative and qualitative MRI was normal. The syndrome in these three kindreds is distinct from temporal lobe epilepsy due to mesial temporal sclerosis and febrile convulsions and is probably a form of idiopathic localization related epilepsy. Its relation to other familial temporal lobe epilepsy phenotypes is discussed. PMID- 11888257 TI - Coping with the challenge of transition in older adolescents with epilepsy. AB - Chronic illnesses, such as epilepsy, have been shown to have detrimental effects on both psychological adjustment and coping behaviour. Using the process model of coping, these effects were investigated in a patient group of 36, 16-21 year olds with epilepsy and a control group of 31 of their peers. Participants completed a postal questionnaire containing measures of psychological adjustment (self esteem, affect, self-efficacy) and an adolescent coping questionnaire. Comparison of the two groups showed that the patient group exhibited significantly more non productive coping than the control group. The control group exhibited significantly more problem solving coping and displayed significantly more problem solving bias than the patient group. No significant differences were found between the patient and control group on measures of psychological adjustment. However, psychological adjustment was found to be associated with coping response in the patient but not the control group. PMID- 11888256 TI - Neuropsychological performance in frontal lobe epilepsy. AB - The search for a special neuropsychological profile of frontal lobe epilepsy subjects (FLE) has so far led to inconclusive results. In this paper we compared the preoperative neuropsychological performance of FLE and temporal lobe epilepsy (TLE) subjects. We further investigated whether frontal lobe lesions of epileptogenic cause produce the same type of cognitive dysfunction as do tumours of the frontal lobe. Sixteen FLE subjects were compared to 16 TLE subjects as well as to a group of 10 subjects after the removal of frontal lobe tumors (TUM) and a healthy control group. A set of neuropsychological test measures routinely used for presurgical evaluation, an emotional conceptualization task and two associative learning tasks were administered. We found that subjects with frontal lobe damage were significantly impaired relative to controls on a wide range of cognitive functions independent of neurological cause. FLE subjects could hardly be discriminated from TLE subjects as both groups showed a similarly reduced level of neuropsychological performance. Our results demonstrate the devastating effect that frontal lobe epilepsy can have on cognitive functioning. Routinely used neuropsychological test measures lack the specificity to distinguish between frontal and temporal lobe epilepsy. Highly specialized measures are necessary to reveal differences. PMID- 11888258 TI - Knowledge, attitude, and practice of epilepsy in rural Sri Lanka. AB - Knowledge, attitude, and practice in relation to epilepsy in developing countries appears to be different from that in developed countries. This study was conducted to evaluate knowledge, attitudes, expectations, sociocultural aspects, patient characteristics, disease characteristics, pattern of drug therapy, and outcome of patients with epilepsy in rural Sri Lanka. Data were collected from 207 patients attending an epilepsy clinic. In general the study shows a positive trend in knowledge, expectations and attitude toward epilepsy. Social morbidity is reported from 53.6% indicating that public attitude towards epilepsy needs to be changed. Alternative modes of treatment have been tried by 45.9%, reflecting the cultural beliefs in the society. 75% are on monotherapy and carbamazepine is the most commonly used drug. Seizure control is excellent (no seizures during the preceding 6 months) in 33.8%. Side effects of antiepileptic drugs are reported by 76.3%. Various kinds of medical morbidity have been experienced by 32.9% of patients. PMID- 11888259 TI - Indicators of inflammation after recent tonic-clonic epileptic seizures correlate with plasma interleukin-6 levels. AB - Cerebrospinal fluid (CSF) and peripheral blood pleocytosis have been observed after epileptic seizures without any evidence of infections, but no systematic studies on the acute phase reaction in such patients have been performed. We have previously reported increased levels of interleukin-6 (IL-6) in patients with recent tonic--clonic seizures. Because IL-6 is a major inducer of the systemic acute phase reaction, we decided to study various indicators of inflammation in the blood as well as their correlation with plasma and CSF IL-6 levels. CSF and blood samples were studied from 37 patients with previously undiagnosed and untreated tonic-clonic seizures without any clinical evidence of systemic or central nervous system infections as well as from 40 controls. The mean peripheral blood and CSF-leukocyte counts were significantly higher in patients compared with controls ( 7.9 x 10(9)vs. 6.1 x 10(9), P= 0.002 and 1.9 x 10(6)vs. 1.1 x 10(6), P= 0.032, respectively). There was some indication of increased concentration of C-reactive protein (CRP) and no difference in haptoglobin levels. There was a significant correlation between plasma but not CSF IL-6 concentration and those of both B-leukocyte count ( r= 0.051, P= 0.009) and CRP ( r= 0.42, P= 0.009). Epileptic seizures provoke a production of cytokines such as IL-6 that may in turn cause an activation of the acute phase reaction. Thus, CSF pleocytosis and increase in some indicators of inflammation should not automatically be attributed to systemic or CNS infections in patients with acute seizures. PMID- 11888260 TI - Role of topiramate in adults with intractable epilepsy, mental retardation, and developmental disabilities. AB - The efficacy and safety of topiramate in patients with intractable mixed seizures, mental retardation (MR), and developmental disabilities (DD) were investigated. Twenty patients (eight females and 12 males) aged 21-57 years old with intractable epilepsy with mixed seizures, MR [profound (five), severe (three), moderate (two), mild (eight) and borderline (two)], and DD were treated with adjunctive topiramate 25 mg per day for 1 week followed by titration to clinical response (range 50-350 mg per day). Other antiepileptic drugs (AEDs) were decreased simultaneously. Topiramate therapy was discontinued in four patients for adverse events consisting of disorientation, unsteadiness, and pneumonia (one patient); anaphylactic shock from a tuna fish allergy (one); patient choice (one); and loss to follow-up (one). Seizures improved by gt-or equal, slanted 50% in 11 of 16 patients (69%). Two patients (13%) were seizure free, including one patient who prior to topiramate therapy was seizure free but experiencing an intolerable adverse effect during therapy with another AED. Seizure duration and/or severity decreased in seven patients (44%). An increase in alertness was observed in 11 patients (59%). Topiramate was associated with improvement in seizure severity and alertness in this series and may be useful as adjunctive therapy in patients with mixed seizures, MR, and DD. PMID- 11888261 TI - Comparative bioethics in bipolar and epilepsy research. AB - RATIONALE: AEDs are increasingly evaluated for efficacy in bipolar disorders utilizing double-blind, placebo-controlled, randomized clinical trials (RCTs) as required by the FDA. However, the risk to patients is under-estimated in trial design. Bipolar depression has a significant risk for suicide; bipolar episodes can lead to kindling with increased long-term morbidity; rapid regression may occur during the placebo phase or during dose ranging trials with resultant active suicide status. The associated risks mandate that the ethics of FDA required protocols are addressed. METHOD: Comparative analysis and literature review of bipolar and epilepsy research designs. RESULTS: In psychiatry, all INDs require RCTs for approval. In epilepsy, AEDs are initially approved as add-on agents only. Once AEDs have demonstrated add-on efficacy, cross-over studies comparing active AEDs, sub-optimal dosing paradigms, new-onset, and pre-surgical inpatient placebo trials are utilized to prove efficacy of the new AED in monotherapy. Ethical considerations to avoid seizures and to minimize risks to subjects have led to newer clinical trial designs. CONCLUSIONS: The FDA initially requires add-on studies with new AEDs due to the risk of seizures during the placebo phase. The author argues that bipolar research warrants similar add-on studies to prove efficacy because the risk of suicide and increased long-term morbidity in the bipolar population is as significant as the risk of seizures in the epilepsy population. Although the number of patients needed to prove statistical efficacy would increase, the safety of such research would also markedly increase. The author further concludes that with the risk of suicide during bipolar research, ethical considerations require increased frequency of patient contact with a significant other co-signing the informed consent for research and serving as a contact for the coordinator. PMID- 11888262 TI - Non-convulsive status epilepticus induced by tiagabine in a patient with pseudoseizure. AB - Tiagabine, a novel GABA reuptake inhibitor, has been reported to induce non convulsive status epilepticus (NCSE) in patients with epilepsy. We report a 27 year old female with history of pseudoseizure documented by video-EEG monitoring who presented confusion while on 56 mg per day of tiagabine. Electroencephalography showed generalized sharp and slow wave discharges, consistent with NCSE. The NCSE was terminated by lorazepam and did not recur after tiagabine was discontinued. This case report suggests that tiagabine may induce NCSE in patients without epilepsy. PMID- 11888263 TI - Temporal lobe epilepsy associated with hippocampal sclerosis and a contralateral middle fossa arachnoid cyst. AB - We report on a 13-year-old boy with temporal lobe epilepsy associated with left hippocampal sclerosis and a contralateral arachnoid cyst in the middle cranial fossa (ACMCF). Chronic intracranial recording from subdural grid electrodes showed the left medial temporal lobe to be the ictal onset zone. After left anterior temporal lobectomy with hippocampectomy, seizure control was improved. ACMCF was not considered the direct cause of epilepsy; instead the seizures were attributed to hippocampal sclerosis. PMID- 11888264 TI - Reflex seizures and non-ketotic hyperglycemia: an unresolved issue. AB - Reflex seizures are a rare form of epilepsy, the pathogenesis of which is unclear. They have been reported in the setting of non-ketotic hyperglycemia (NKH) and are considered to be neuroendocrine in origin. We report a diabetic patient with movement-induced seizures whose presentation suggests that brain ischemia may be the precipitating event in focal seizures seen in the setting of NKH. We recommend that in such instances a focal lesion such as stroke should be ruled out. PMID- 11888265 TI - Seizure-related headache in patients with epilepsy. AB - We investigated the type and frequency of interictal primary headache and peri ictal headache in 109 patients with partial-onset and 26 patients with generalized onset seizures in this study. Interictal headaches were present in 50 (40.7%) of 135 patients. Comparing the interictal headache on the basis of seizure type, we couldn't find any significant difference between the seizure groups. Seventy-nine (58.51%) patients had peri-ictal headache. Eleven of these patients had pre-ictal headache (PriH), three of all had ictal headache and, 56 of these had post-ictal headache (PoiH). PriH and PoiH were more frequently encountered before and after secondary generalized tonic-clonic seizures (GTCS) compared to other seizure groups. The type of pain in PoiH was 'throbbing' in complex partial seizures and 'steady' in GTCS. PMID- 11888266 TI - Oxcarbazepine. PMID- 11888267 TI - Epilepsy neurosurgery: a patient's perspective. AB - Contemplation of epilepsy neurosurgery for a patient with lifetime epilepsy poses difficult decisions and can be quite fearful. As epileptologists, we often do not appreciate the degree of patients' concerns, especially whether quality of life could be worsened by surgical complications. In this poem, a patient lyrically describes this dilemma. PMID- 11888268 TI - Conformation and stability of alpha-helical membrane proteins. 1. Influence of salts on conformational equilibria between active and Inactive states of rhodopsin. AB - We studied the influence of salts on the pH-dependent conformational equilibria between the active and the inactive photoproduct states of rhodopsin, Meta II and Meta I, respectively, and between the active and inactive conformations of the apoprotein opsin. In both equilibria, the active species is favored in the presence of medium to high concentration of salt. The ion selectivity for the Meta I/Meta II equilibrium is particularly pronounced for the anions and follows the series trichloroacetate > thiocyanate > iodide > bromide > sulfate > chloride > acetate. The Hill coefficient of this salt-induced transition is close to 2.0. Both ion selectivity and Hill coefficient suggest that the transition is mainly regulated by ion binding to two specific charged binding sites in the protein with smaller contributions being due to the Hofmeister effect. We propose that these putative ion binding sites are identical to those sites that are titrated in the corresponding pH-dependent conformational transition. They presumably function as ionic locks, which keep the receptor in an inactive conformation, and which may be disrupted either by pH-dependent protonation or by salt-dependent ion binding. PMID- 11888269 TI - Conformation and stability of alpha-helical membrane proteins. 2. Influence of pH and salts on stability and unfolding of rhodopsin. AB - We studied the stability and pH-induced denaturation of rhodopsin and its photoproducts as a model for alpha-helical membrane proteins. The increased stability of the dark state of rhodopsin as compared to its photoproduct states allows the initiation of unfolding of the protein by light-dependent isomerization of the chromophore. We could therefore characterize the transition from the native to either acid or alkaline denatured states by light-induced Fourier transform infrared difference spectroscopy, UV-visible spectroscopy, and intrinsic tryptophan fluorescence spectroscopy. The results indicate a loss of important tertiary interactions within the protein and between the protein and the retinal chromophore in the denatured state, despite that the secondary structure of the protein is almost fully retained during the transition. We therefore propose that in this denatured state the protein adopts the conformation of a loose bundle of preserved, but only weakly interacting, transmembrane helices with a largely des-oriented and partly solvent-exposed chromophore. We further characterized the influence of salts on the stability of the rhodopsin helix bundle, which was found to follow the Hofmeister series. We found that the effect of sodium chloride may be stabilizing or destabilizing, depending on the intrinsic stability of the examined protein conformation and on salt concentration. In particular, sodium chloride is shown to counteract the formation of the denatured loose bundle state presumably by increasing the lateral pressure on the helix bundle, thereby stabilizing native-like tertiary contacts within the protein. PMID- 11888270 TI - Metal ion-induced stabilization and refolding of anticoagulation factor II from the venom of Agkistrodon acutus. AB - Anticoagulation factor II (ACF II) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X-binding protein in a Ca(2+)-dependent fashion with marked anticoagulant activity. The equilibrium unfolding/refolding of apo-ACF II, holo-ACF II, and Tb(3+)-reconstituted ACF II in guanidine hydrochloride (GdnHCl) solutions was studied by following the fluorescence and circular dichroism (CD). Metal ions were found to increase the structural stability of ACF II against GdnHCl and irreversible thermal denaturation and, furthermore, influence its unfolding/refolding behavior. The GdnHCl-induced unfolding/refolding of both apo-ACF II and Tb(3+)-ACF II is a two-state process with no detectable intermediate state, while the GdnHCl-induced unfolding/refolding of holo-ACF II in the presence of 1 mM Ca(2+) follows a three state transition with an intermediate state. Ca(2+) ions play an important role in the stabilization of both native and I states of holo-ACF II. The decalcification of holo-ACF II shifts the ending zone of unfolding/refolding curve toward lower GdnHCl concentration, while the reconstitution of apo-ACF II with Tb(3+) ions shifts the initial zone of the denaturation curve toward higher GdnHCl concentration. Therefore, it is possible to find a denaturant concentration (2.1 M GdnHCl) at which refolding from the fully denatured state of apo-ACF II to the I state of holo-ACF II or to the native state of Tb(3+)-ACF II can be initiated merely by adding the 1 mM Ca(2+) ions or 10 microM Tb(3+) ions to the unfolded state of apo-ACF II, respectively, without changing the concentration of the denaturant. Using Tb(3+) as a fluorescence probe of Ca(2+), the kinetic results of metal ion-induced refolding provide evidence for the fact that the first phase of Tb(3+)-induced refolding should involve the formation of the compact metal-binding site regions, and subsequently, the protein undergoes further conformational rearrangements to form the native structure. PMID- 11888272 TI - Mapping nucleosome locations on the 208-12 by AFM provides clear evidence for cooperativity in array occupation. AB - Concatameric sea urchin 5S rDNA templates reconstituted with histones provide very popular chromatin models for many kinds of in vitro studies. We have used AFM to characterize the locational aspects of nucleosome occupation on one such array, the 208-12, by determining the internucleosomal- and end-distance distributions for arrays reconstituted to various subsaturating levels with nonacetylated or hyperacetylated HeLa histones. A simulation analysis of the experimental distributions confirms the qualitative conclusions and provides quantitative parameter values for the identified features. For nonacetylated arrays, the end-distance data demonstrate the nucleosome positioning ability of the 5S sequence and detect an enhanced preference for nucleosomes to bind at DNA termini. The internucleosomal-distance data provide clear evidence for cooperativity in nucleosome location on these templates, detectable even at subsaturated loading levels. Hyperacetylated arrays show no change in the preference of nucleosomes to bind at termini and a slight change in nucleosome positioning behavior but, most strikingly, little or no evidence for cooperativity in nucleosome location. Thus, acetylation of the N-terminal histone tails abolishes the cooperativity. PMID- 11888271 TI - Kinetic and structural studies on the interaction of cholinesterases with the anti-Alzheimer drug rivastigmine. AB - Rivastigmine, a carbamate inhibitor of acetylcholinesterase, is already in use for treatment of Alzheimer's disease under the trade name of Exelon. Rivastigmine carbamylates Torpedo californica acetylcholinesterase very slowly (k(i) = 2.0 M( 1) min(-1)), whereas the bimolecular rate constant for inhibition of human acetylcholinesterase is >1600-fold higher (k(i) = 3300 M(-1) min(-1)). For human butyrylcholinesterase and for Drosophila melanogaster acetylcholinesterase, carbamylation is even more rapid (k(i) = 9 x 10(4) and 5 x 10(5) M(-1) min(-1), respectively). Spontaneous reactivation of all four conjugates is very slow, with <10% reactivation being observed for the Torpedo enzyme after 48 h. The crystal structure of the conjugate of rivastigmine with Torpedo acetylcholinesterase was determined to 2.2 A resolution. It revealed that the carbamyl moiety is covalently linked to the active-site serine, with the leaving group, (-)-S-3-[1 (dimethylamino)ethyl]phenol, being retained in the "anionic" site. A significant movement of the active-site histidine (H440) away from its normal hydrogen-bonded partner, E327, was observed, resulting in disruption of the catalytic triad. This movement may provide an explanation for the unusually slow kinetics of reactivation. PMID- 11888274 TI - Kinetic and thermodynamic basis of promoter strength: multiple steps of transcription initiation by T7 RNA polymerase are modulated by the promoter sequence. AB - Transcription initiation by T7 RNA polymerase (T7 RNAP) is regulated by the specific promoter DNA sequence that is classically divided into two major domains, the binding domain (-17 to -5) and the initiation domain (-4 to +6). The occurrence of non-consensus bases within these domains is responsible for the diversity of promoter strength, the basis of which was investigated by studying T7 promoters with changes in the promoter specificity region (-13 to -6) of the binding domain and/or the melting region (-4 to -1) of the initiation domain. The transient state kinetics and thermodynamic studies revealed that multiple steps in the pathway of transcription initiation are modulated by the promoter DNA sequence. Three base changes in the promoter specificity region at -11, -12, and 13, found in the natural phi 3.8 promoter, reduced the overall affinity of the T7 RNAP for the promoter DNA by 2-3-fold and decreased the rate of pppGpG synthesis, the first RNA product. Promoter opening is thermodynamically driven in T7 RNAP, and a single base change in the melting region (TATA to TAAA) decreased the extent of open complex generated at equilibrium. This base change in the melting region also increased the K(d) of (+1) GTP and the dissociation rate of pppGpG. Thus, transcription initiation at various T7 promoters is differentially regulated by initiating GTP concentration. The specificity and melting regions of T7 promoter DNA act both independently and synergistically to affect distinct steps of transcription initiation. Although each step in the initiation pathway is affected to a small degree by promoter sequence variations, the cumulative effect dictates the overall promoter strength. PMID- 11888273 TI - The human interferon receptor: NMR-based modeling, mapping of the IFN-alpha 2 binding site, and observed ligand-induced tightening. AB - The human interferon receptor (IFNAR) mediates the antiviral and antiproliferative activities of type I interferons (IFNs). This receptor is comprised of subunits IFNAR1 and IFNAR2, the latter exhibiting nanomolar affinity for IFNs. Here the extracellular domain of IFNAR2 (IFNAR2-EC), a soluble 25 kDa IFN-binding polypeptide, and its complex with IFN-alpha 2 were studied using multidimensional NMR. IFNAR2-EC is comprised of two fibronectin-III (FN-III) domains connected by a helical hinge region. The deduced global fold was utilized to improve the alignment of IFNAR2-EC against structurally related receptors and to model its structure. A striking feature of IFNAR2-EC is the limited and localized deviations in chemical shifts exhibited upon ligand binding, observed for only 15% of its backbone (1)H and (15)N nuclei. Analysis of these deviations maps the IFN-alpha 2 binding site upon IFNAR2-EC to a contiguous surface on the N terminal domain, including the S3-S4 loop (residues 44-53), the S5-S6 loop and S6 beta-strand (residues 74-82), and the S7 beta-strand and the hinge region (residues 95-105). The C-terminal domain contributes only marginally to ligand binding, and no change in the hypothesized interdomain interface is observed. The proposed binding domain encompasses all residues implicated by mutagenesis studies in IFN binding, and suggests adjacent residues cooperate in forming the binding surface. D(2)O-exchange experiments indicate that binding of IFN-alpha2 induces tightening of the N-terminal domain of IFNAR2-EC. This increase in receptor rigidity may play an important role in initiating the intracellular stage of the IFN signaling cascade. PMID- 11888275 TI - NMR structure of the second intracellular loop of the alpha 2A adrenergic receptor: evidence for a novel cytoplasmic helix. AB - A major, unresolved question in signal transduction by G protein coupled receptors (GPCRs) is to understand how, at atomic resolution, a GPCR activates a G protein. A step toward answering this question was made with the determination of the high-resolution structure of rhodopsin; we now know the intramolecular interactions that characterize the resting conformation of a GPCR. To what degree does this structure represent a structural paradigm for other GPCRs, especially at the cytoplasmic surface where GPCR-G protein interaction occurs and where the sequence homology is low among GPCRs? To address this question, we performed NMR studies on approximately 35-residue-long peptides including the critical second intracellular loop (i2) of the alpha 2A adrenergic receptor (AR) and of rhodopsin. To stabilize the secondary structure of the peptide termini, 4-12 residues from the adjacent transmembrane helices were included and structures determined in dodecylphosphocholine micelles. We also characterized the effects on an alpha 2A AR peptide of a D130I mutation in the conserved DRY motif. Our results show that in contrast to the L-shaped loop in the i2 of rhodopsin, the i2 of the alpha 2A AR is predominantly helical, supporting the hypothesis that there is structural diversity within GPCR intracellular loops. The D130I mutation subtly modulates the helical structure. The spacing of nonpolar residues in i2 with helical periodicity is a predictor of helical versus loop structure. These data should lead to more accurate models of the intracellular surface of GPCRs and of receptor-mediated G protein activation. PMID- 11888276 TI - Mechanism of domain closure of Sec7 domains and role in BFA sensitivity. AB - Activation of small G proteins of the Arf family is initiated by guanine nucleotide exchange factors whose catalytic Sec7 domain stimulates the dissociation of the tightly bound GDP nucleotide. The exchange reaction involves distinct sequential steps that can be trapped by the noncompetitive inhibitor brefeldin A, by mutation of an invariant catalytic glutamate, or by removal of guanine nucleotides. Arf-GDP retains most characteristics of its GDP-bound form at the initial low-affinity Arf-GDP-Sec7 step. It then undergoes large conformational changes toward its GTP-bound form at the next step, and eventually dissociates GDP to form a nucleotide-free high-affinity Arf-Sec7 complex at the last step. Thus, Arf proteins evolve through different conformations that must be accommodated by Sec7 domains in the course of the reaction. Here the contribution of the flexibility of Sec7 domains to the exchange reaction was investigated with the crystal structure of the unbound Sec7 domain of yeast Gea2. Comparison with Gea2 in complex with nucleotide-free Arf1 Delta 17 [Goldberg, J. (1998) Cell 95, 237-248] reveals that Arf induces closure of the two subdomains that form the sides of its active site. Several residues that determine sensitivity to brefeldin A are involved in interdomain and local movements, pointing to the importance of the flexibility of Sec7 domains for the inhibition mechanism. Altogether, this suggests a model for the initial steps of the exchange reaction where Arf docks onto the C-terminal domain of the Sec7 domain before closure of the N-terminal domain positions the catalytic glutamate to complete the reaction. PMID- 11888277 TI - The N-terminal extension of rusticyanin is not responsible for its acid stability. AB - The N-terminal extension of rusticyanin is a unique structural feature of this protein in the cupredoxin family and has been speculated to be responsible for the extreme acid stability of the protein. We have removed the 35 residues from the N-terminus and show that the resulting -35 mutant is insoluble in aqueous media above pH 5.0 and exists primarily in a hexameric form at lower pHs. Synchrotron radiation circular dichroism (SRCD) and solution X-ray scattering data indicate that much of the beta-sheet structure is retained in acidic solution and indeed there is a small but significant increase in the beta-sheet contribution. We suggest this to be a result of beta-sheet formation between the monomer interfaces. The mutant does not bind copper. These results provide evidence that the unique N-terminus of rusticyanin is not responsible for the acid stability of the hydrophobic beta-barrel core of the protein. PMID- 11888279 TI - Biophysical properties of camelid V(HH) domains compared to those of human V(H)3 domains. AB - Camelidae possess an unusual form of antibodies lacking the light chains. The variable domain of these heavy chain antibodies (V(HH)) is not paired, while the V(H) domain of all other antibodies forms a heterodimer with the variable domain of the light chain (V(L)), held together by a hydrophobic interface. Here, we analyzed the biophysical properties of four camelid V(HH) fragments (H14, AMD9, RN05, and CA05) and two human consensus V(H)3 domains with different CDR3 loops to gain insight into factors determining stability and aggregation of immunoglobulin domains. We show by denaturant-induced unfolding equilibria that the free energies of unfolding of V(HH) fragments are characterized by Delta G(N U) values between 21.1 and 35.0 kJ/mol and thus lie in the upper range of values for V(H) fragments from murine and human antibodies. Nevertheless, the V(HH) fragments studied here did not reach the high values between 39.7 and 52.7 kJ/mol of the human consensus V(H)3 domains with which they share the highest degree of sequence similarity. Temperature-induced unfolding of the V(HH) fragments that were studied proved to be reversible, and the binding affinity after cooling was fully retained. The melting temperatures were determined to be between 60.1 and 66.7 degrees C. In contrast, the studied V(H)3 domains aggregated during temperature-induced denaturation at 63-65 degrees C. In summary, the camelid V(HH) fragments are characterized by a favorable but not unusually high stability. Their hallmark is the ability to reversibly melt without aggregation, probably mediated by the surface mutations characterizing the V(HH) domains, which allow them to regain binding activity after heat renaturation. PMID- 11888278 TI - Function of extracellular loop 2 in rhodopsin: glutamic acid 181 modulates stability and absorption wavelength of metarhodopsin II. AB - The second extracellular loop of rhodopsin folds back into the membrane-embedded domain of the receptor to form part of the binding pocket for the 11-cis retinylidene chromophore. A carboxylic acid side chain from this loop, Glu181, points toward the center of the retinal polyene chain. We studied the role of Glu181 in bovine rhodopsin by characterizing a set of site-directed mutants. Sixteen of the 19 single-site mutants expressed and bound 11-cis-retinal to form pigments. The lambda(max) value of mutant pigment E181Q showed a significant spectral red shift to 508 nm only in the absence of NaCl. Other substitutions did not significantly affect the spectral features of the mutant pigments in the dark. Thus, Glu181 does not contribute significantly to spectral tuning of the ground state of rhodopsin. The most likely interpretation of these data is that Glu181 is protonated and uncharged in the dark state of rhodopsin. The Glu181 mutants displayed significantly increased reactivity toward hydroxylamine in the dark. The mutants formed metarhodopsin II-like photoproducts upon illumination but many of the photoproducts displayed shifted lambda(max) values. In addition, the metarhodopsin II-like photoproducts of the mutant pigments had significant alterations in their decay rates. The increased reactivity of the mutants to hydroxylamine supports the notion that the second extracellular loop prevents solvent access to the chromophore-binding pocket. In addition, Glu181 strongly affects the environment of the retinylidene Schiff base in the active metarhodopsin II photoproduct. PMID- 11888280 TI - Role of the N-terminal helix I for dimerization and stability of the calcium binding protein S100B. AB - Human S100B(beta beta) is a small intracellular EF-hand calcium-binding protein that consists of two noncovalently associated 91-residue beta monomers. The three dimensional structures of S100B reveal the dimer interface consists of four alpha helices (I, I' and IV, IV') packed in an X-type bundle. In this study, guanidine hydrochloride denaturation and dynamic light scattering were used to assess the impact of single (L3A, L3S, M7A, I11A, F14A) and double (L3A/I11A and L3A/F14A) substitution mutations in helix I on the stability and dimerization propensity of S100B. The free energy of unfolding (Delta G(u)) of wild-type apo-S100B was determined to be 72.4 +/- 4.0 kJ mol(-1), consistent with it being the most stable calcium-binding protein to date. The order of stability of the mutants in their apo form is S100B > L3A > L3S > I11A > M7A approximately L3A/I11A > F14A > L3A/F14A. Further, there is a strong correlation between the stability and the cooperativity of unfolding. Each mutation proved to be more stable in its calcium form compared to its apo form. The calcium-bound L3S substitution proved to be significantly more stable than calcium-saturated S100B, whereas the L3A, I11A, and L3A/I11A mutants are only slightly more stable than the wild-type protein. The F14A and L3A/F14A mutants are significantly reduced in stability, even in the presence of calcium. PMID- 11888281 TI - Polar residues in membrane domains of proteins: molecular basis for helix-helix association in a mutant CFTR transmembrane segment. AB - Polar side chains constitute over 20% of residues in the transmembrane (TM) helices of membrane proteins, where they may serve as hydrogen bond interaction sites for phenotypic polar mutations that arise in membrane protein-related diseases. To systematically explore the structural consequences of H-bonds between TM helices, we focused on TM4 of the cystic fibrosis conductance regulator (CFTR) and its cystic fibrosis- (CF-) phenotypic mutation, V232D, as a model system. Synthetic peptides corresponding to wild-type (TM4-wt) (residues 219-242: LQASAFCGLGFLIVLALFQAGLGR) and mutant (TM4-V232D) sequences both adopt helical structures in SDS micelles and display dimer bands on SDS-PAGE arising from disulfide bond formation via wild-type residue Cys-225. However, the TM4 V232D peptide additionally forms a ladder of noncovalent oligomers, including tetramers, hexamers, and octamers, mediated by a hydrogen bond network involving Asp-Gln side chain-side chain interactions. Ala-scanning mutagenesis of the TM4 sequence indicated that ladder formation minimally required the simultaneous presence of the Cys-225, Asp-232, and Gln-237 residues. As random hydrophobic sequences containing these three residues at TM4 equivalent positions did not oligomerize, specific van der Waals packing interactions between helix side chains were also shown to play a crucial role. Overall, the results suggest that polar mutations in membrane domains, in conjunction with critically positioned polar partner residues, potentially constitute a source of aberrant helix interactions that could contribute to loss of function when they arise in protein transmembrane domains. PMID- 11888282 TI - ATP hydrolysis induces expansion of MutS contacts on heteroduplex: a case for MutS treadmilling? AB - An unsolved problem in E. coli mismatch repair is how the MutS-MutL complex communicates positional information of a mismatch to MutH. MutS is bound to a mismatch in the absence of ATP, exhibiting a short DNase I footprint that is dramatically expanded in ATP hydrolysis. The same is corroborated by restriction enzyme site protection far away from the mismatch. High-resolution gel-shift analyses revealed that super-shifted specific complexes, presumably containing multiple MutS homodimers on the same heteroduplex, were generated during ATP hydrolysis. Such complexes are largely nonspecific in "minus ATP" or in ATP gamma S conditions. Specific ternary complexes of MutS-MutL-heteroduplexes were formed only during ATP hydrolysis. These results suggest that MutS loading onto a mismatch induces the formation of a higher-order complex containing multiple MutS homodimers, presumably through a putative "treadmilling action" that is ATP hydrolysis dependent. Such a higher-order MutS complex productively interacts with MutL in ATP-hydrolyzing conditions and generates a specific ternary complex, which might communicate with MutH. This model should neither depend on nor give rise to the spooling of DNA. This was corroborated when we observed footprint extension in ATP-hydrolyzing conditions, despite the heteroduplex ends being tethered to agarose beads that block helical rotations. PMID- 11888283 TI - pK(a) perturbation in genomic Hepatitis Delta Virus ribozyme catalysis evidenced by nucleotide analogue interference mapping. AB - The Hepatitis Delta Virus (HDV) ribozyme was the first RNA enzyme proposed to use a proton-transfer mechanism for catalysis. Previous biochemical evidence suggested that the genomic HDV ribozyme promotes cis-cleavage using cytosine 75 whose pK(a) is perturbed within the active site. Here we present further biochemical evidence for the involvement of C75 in proton transfer, as well as evidence to support a plausible mechanism for C75 pK(a) perturbation. Nucleotide analogue interference mapping (NAIM) experiments with C analogues having altered N3 pK(a)s demonstrate the importance of C75 ionization in the HDV cis-cleavage reaction. pH-dependent interference rescue with C analogues having enhanced N3 acidity indicates that C75 is the only cytidine residue that must be protonated for ribozyme activity. Furthermore, interference analysis with pseudoisocytidine, a charge-neutral mimic of a C with a protonated N3, shows a pattern consistent with proton transfer, possibly from the C75 N3 to the 5'-oxyanion leaving group during the cis-cleavage reaction. Strong pH-independent inhibition of ribozyme function also occurs at C75 with a C analogue that lacks the N4 amino group, implicating the exocyclic amine in critical interactions in the active site. Interactions with the amino group may play an important role in perturbing the C75 N3 pK(a). Protonation of C41 has been proposed to be important for ribozyme activity; however, no interference at C41 was observed in this analogue series, which argues against a functional role for C41 protonation. These data support a model wherein C75 of the genomic HDV ribozyme acts as a general acid during its cis-cleavage reaction, and provide a glimpse into how RNAs, in a manner similar to protein enzymes, might employ local environmental electronic modulation to catalyze reactions. PMID- 11888284 TI - Antiproliferative activity of G-quartet-forming oligonucleotides with backbone and sugar modifications. AB - Oligonucleotide-based therapies have considerable potential in cancer, viral, and cardiovascular disease therapies. However, it is becoming clear that the biological effects of oligonucleotides are not solely due to the intended sequence-specific interactions with nucleic acids. Oligonucleotides are also capable of interacting with numerous cellular proteins owing to their polyanionic character or specific secondary structure. We have examined the antiproliferative activity, protein binding, and G-quartet formation of a series of guanosine-rich oligonucleotides, which are analogues of GRO29A, a G-quartet forming, growth inhibitory oligonucleotide, whose effects we have previously described [Bates P. J., Kahlon, J. B., Thomas, S. D., Trent, J. O., and Miller, D. M. (1999) J. Biol. Chem. 274, 26369-26377]. The GRO29A analogues include phosphorothioate (PS29A), 2'-O-methyl RNA (MR29A), and mixed DNA/2'-O-methyl RNA (MRdG29A) oligonucleotides. We demonstrate by UV spectroscopy that all of the modified analogues form stable structures, which are consistent with G-quartet formation. We find that the phosphorothioate and mixed DNA/2'-O-methyl analogues are able to significantly inhibit proliferation in a number of tumor cell lines, while the 2' O-methyl RNA has no significant effects. Similar to the original oligonucleotide, GRO29A, the growth inhibitory oligonucleotides were able to compete with the human telomere sequence oligonucleotide for binding to a specific cellular protein. The less active MR29A does not compete significantly for this protein. On the basis of molecular modeling of the oligonucleotide structures, it is likely that the inactivity of MR29A is due to the differences in the groove structure of the quadruplex formed by this oligonucleotide. Interestingly, all GRO29A analogues, including an unmodified DNA phosphodiester oligonucleotide, are remarkably resistant to nuclease degradation in the presence of serum-containing medium, indicating that secondary structure plays an important role in biological stability. The remarkable stability and strong antiproliferative activity of these oligonucleotides confirm their potential as therapeutic agents. PMID- 11888285 TI - Peptide nucleic acid (PNA) facilitates multistranded hybrid formation between linear double-stranded DNA targets and RecA protein-coated complementary single stranded DNA probes. AB - RecA protein-coated single-stranded DNA probes, known as RecA nucleoprotein filaments, bind specifically to homologous DNA sequences within double-stranded DNA targets, forming multistranded probe-target DNA hybrids. This DNA hybridization reaction can be used for sequence-specific gene capture, gene modification, and gene regulation. Thus, factors that enhance the efficiency of the hybridization reaction are of significant practical importance. We show here that the hybridization of a peptide nucleic acid (PNA) within or adjacent to the probe-target homology region significantly enhances the yield of hybrid DNA formed in the reaction between linear double-stranded DNA targets and RecA protein-coated complementary single-stranded (css)DNA probes. The possible mechanisms and the advantages of using RecA nucleoprotein filaments in combination with PNA for genomic DNA cloning and mutagenesis are presented. PMID- 11888286 TI - Cooperative binding of ATP and RNA substrates to the DEAD/H protein DbpA. AB - Unlike most DEAD/H proteins, the purified Escherichia coli protein DbpA demonstrates high specificity for its 23S rRNA substrate in vitro. Here we describe several assays designed to characterize the interaction of DbpA with its RNA and ATP substrates. Electrophoretic mobility shift assays reveal a sub nanomolar binding affinity for a 153 nucleotide RNA substrate (R153) derived from the 23S rRNA. High affinity RNA binding requires both hairpin 92 and helix 90, as substrates lacking these structures bind DbpA with lower affinity. AMPPNP inhibition assays and ATP/ADP binding assays provide binding constants for ATP and ADP to DbpA with and without RNA substrates. These data have been used to describe a minimal thermodynamic scheme for the binding of the RNA and ATP substrates to DbpA, which reveals cooperative binding between larger RNAs and ATP with cooperative energies of approximately 1.3 kcal mol(-1). This cooperativity is lost upon removal of helix 89 from R153, suggesting this helix is either the preferred target for DbpA's helicase activity or is a necessary structural element for organization of the target site within R153. PMID- 11888287 TI - Mass spectrometric assays for the tandem lesion 8,5'-cyclo-2'-deoxyguanosine in mammalian DNA. AB - 8,5'-Cyclopurine 2'-deoxynucleosides are among the major lesions in DNA that are formed by attack of hydroxyl radical. These compounds represent a concomitant damage to both sugar and base moieties of the same nucleoside and thus can be considered tandem lesions. Because of the presence of a covalent bond between the sugar and purine moieties, these tandem lesions are not repaired by base excision repair but by nucleotide excision repair. Thus, they may play a role in diseases with defective nucleotide excision repair. We recently reported the identification and quantification of 8,5'-cyclo-2'-deoxyadenosine (8,5'-cdAdo) in DNA by liquid chromatography/mass spectrometry with the isotope dilution technique (LC/IDMS) [Dizdaroglu, M., Jaruga, P., and Rodriguez, H. (2001) Free Radical Biol. Med. 30, 774-784]. In the present work, we investigated the measurement of 8,5'-cyclo-2'-deoxyguanosine (8,5'-cdGuo) in DNA by LC/IDMS. A methodology was developed for the separation of both (5'R)- and (5'S) diastereomers of this compound in enzymic hydrolysates of DNA. The mass spectra were recorded using an atmospheric pressure ionization-electrospray process in the positive ionization mode. For quantification, stable isotope-labeled analogues of (5'R)-8,5'-cdGuo and (5'S)-8,5'-cdGuo were prepared and isolated by semipreparative LC to be used as internal standards. The sensitivity level of LC/MS in the selected ion monitoring mode (LC/MS-SIM) was determined to be approximately 15 fmol of these compounds on the LC column. The yield of 8,5' cdGuo was measured in DNA exposed in aqueous solution to ionizing radiation at doses from 2.5 to 40 Gy. For comparison, gas chromatography/mass spectrometry with the isotope dilution technique (GC/IDMS) was also employed to measure both (5'R)-8,5'-cdGuo and (5'S)-8,5'-cdGuo in DNA. Both techniques yielded nearly identical results. The radiation chemical yield of 8,5'-cdGuo was similar to those of other major purine-derived lesions in DNA. The sensitivity level of GC/MS-SIM was determined to be significantly greater than that of LC/MS-SIM (1 vs 15 fmol). The background levels of (5'R)-8,5'-cdGuo and (5'S)-8,5'-cdGuo were measured in calf thymus DNA and in DNA samples isolated from three different types of cultured human cells. The levels of (5'R)-8,5'-cdGuo and (5'S)-8,5' cdGuo were approximately 2 lesions/10(6) DNA nucleosides and 10 lesions/10(6) DNA nucleosides, respectively. No significant differences between tissues were observed in terms of these background levels. The results showed that both LC/IDMS and GC/IDMS are well suited for the sensitive detection and precise quantification of both (5'R)-8,5'-cdGuo and (5'S)-8,5'-cdGuo in DNA. PMID- 11888288 TI - Hydrolysis of thionopeptides by the aminopeptidase from Aeromonas proteolytica: insight into substrate binding. AB - A series of L-leucine aniline analogues were synthesized that contained either a carbonyl or thiocarbonyl as a part of the amide bond. Additionally, the para position on the phenyl ring of several substrates was altered with various electron-withdrawing or donating groups. The kinetic constants K(m) and k(cat) were determined for the hydrolysis of each of these compounds in the presence of the aminopeptidase from Aeromonas proteolytica (AAP) containing either Zn(II) or Cd(II). The dizinc(II) form of AAP ([ZnZn(AAP)]) was able to cleave both carbonyl and thiocarbonyl containing peptide substrates with similar efficiency. However, the dicadmium(II) form of AAP ([CdCd(AAP)]) was unable to cleave any of the carbonyl-containing compounds tested but was able to cleave the thionopeptide substrates. This is consistent with the borderline hard/soft nature of Zn(II) vs Cd(II). The trends observed in the K(m) values suggest that the oxygen atom of the amide bond directly interacts with the dinuclear active site of AAP. Heterodimetallic forms of AAP that contained one atom of Zn(II) and one of Cd(II) (i.e., [CdZn(AAP)] and [ZnCd(AAP)]) were also prepared. The K(m) values for the thionopeptides substrates are the smallest when Cd(II) is in the first metal binding site, suggesting that substrate binds to the first metal binding site. 1 Phenyl-2-thiourea (PTU) and urea (PU) were also examined to determine the differences between thionopeptide and peptide binding to AAP. PTU and PU were found to be competitive inhibitors of AAP with inhibition constants of 0.24 and 4.6 mM, respectively. The electronic absorption and EPR spectra of [CoCo(AAP)], [CoZn(AAP)], and [ZnCo(AAP)] were recorded in the absence and presence of both PU and PTU. Spectral changes were observed for PTU binding to [CoCo(AAP)] and [CoZn(AAP)] but not for [ZnCo(AAP)], while no spectral changes were observed for any of the Co(II)-substituted forms of AAP upon the addition of PU. These data indicate that carbonyl binding occurs only at the first metal binding site. In light of the data presented herein, the substrate binding step in the proposed mechanism of AAP catalyzed peptide hydrolysis can be further refined. PMID- 11888289 TI - Production and characterization of bifunctional enzymes. Domain swapping to produce new bifunctional enzymes in the aspartate pathway. AB - The bifunctional enzyme aspartokinase-homoserine dehydrogenase I from Escherichia coli catalyzes non-consecutive reactions in the aspartate pathway of amino acid biosynthesis. Both catalytic activities are subject to allosteric regulation by the end product amino acid L-threonine. To examine the kinetics and regulation of the enzymes in this pathway, each of these catalytic domains were separately expressed and purified. The separated catalytic domains remain active, with each of their catalytic activities enhanced in comparison to the native enzyme. The allosteric regulation of the kinase activity is lost, and regulation of the dehydrogenase activity is dramatically decreased in these separate domains. To create a new bifunctional enzyme that can catalyze consecutive metabolic reactions, the aspartokinase I domain was fused to the enzyme that catalyzes the intervening reaction in the pathway, aspartate semialdehyde dehydrogenase. A hybrid bifunctional enzyme was also created between the native monofunctional aspartokinase III, an allosteric enzyme regulated by lysine, and the catalytic domain of homoserine dehydrogenase I with its regulatory interface domain still attached. In this hybrid the kinase activity remains sensitive to lysine, while the dehydrogenase activity is now regulated by both threonine and lysine. The dehydrogenase domain is less thermally stable than the kinase domain and becomes further destabilized upon removal of the regulatory domain. The more stable aspartokinase III is further stabilized against thermal denaturation in the hybrid bifunctional enzyme and was found to retain some catalytic activity even at temperatures approaching 100 degrees C. PMID- 11888290 TI - Production and characterization of bifunctional enzymes. Substrate channeling in the aspartate pathway. AB - The direct channeling of an intermediate between enzymes that catalyze consecutive reactions in a pathway offers the possibility of an efficient, exclusive, and protected means of metabolite delivery. Aspartokinase-homoserine dehydrogenase I (AK-HDH I) from Escherichia coli is an unusual bifunctional enzyme in that it does not catalyze consecutive reactions. The potential channeling of the intermediate beta-aspartyl phosphate between the aspartokinase of this bifunctional enzyme and aspartate semialdehyde dehydrogenase (ASADH), the enzyme that catalyzes the intervening reaction, has been examined. The introduction of increasing levels of inactivated ASADH has been shown to compete against enzyme-enzyme interactions and direct intermediate channeling, leading to a decrease in the overall reaction flux through these consecutive enzymes. These same results are obtained whether these experiments are conducted with aspartokinase III, a naturally occurring monofunctional isozyme, with an artificially produced monofunctional aspartokinase I, or with a fusion construct of AK I-ASADH. These results provide definitive evidence for the channeling of beta-aspartyl phosphate between aspartokinase and aspartate semialdehyde dehydrogenase in E. coli and suggest that ASADH may provide a bridge to channel the intermediates between the non-consecutive reactions of AK-HDH I. PMID- 11888291 TI - Molecular cloning and characterization of a tumor-associated, growth-related, and time-keeping hydroquinone (NADH) oxidase (tNOX) of the HeLa cell surface. AB - NOX proteins are growth-related cell surface proteins that catalyze both hydroquinone or NADH oxidation and protein disulfide interchange and exhibit prion-like properties. The two enzymatic activities alternate to generate a regular period length of about 24 min. Here we report the expression, cloning, and characterization of a tumor-associated NADH oxidase (tNOX). The cDNA sequence of 1830 bp is located on gene Xq25-26 with an open reading frame encoding 610 amino acids. The activities of the bacterially expressed tNOX oscillate with a period length of 22 min as is characteristic of tNOX activities in situ. The activities are inhibited completely by capsaicin, which represents a defining characteristic of tNOX activity. Functional motifs identified by site-directed mutagenesis within the C-terminal portion of the tNOX protein corresponding to the processed plasma membrane-associated form include quinone (capsaicin), copper and adenine nucleotide binding domains, and two cysteines essential for catalytic activity. Four of the six cysteine to alanine replacements retained enzymatic activity, but the period lengths of the oscillations were increased. A single protein with two alternating enzymatic activities indicative of a time-keeping function is unprecedented in the biochemical literature. PMID- 11888292 TI - Differential inhibition of Hsc70 activities by two Hsc70-binding peptides. AB - The ability of two high-affinity Hsc70-binding peptides [FYQLALT (peptide-Phi) and NIVRKKK (peptide-K)] to differentially inhibit Hsc70-dependent processes in rabbit reticulocyte lysate (RRL) was examined. Both peptide-Phi and peptide-K inhibited chaperone-dependent renaturation of luciferase in RRL. Peptide-Phi, but not peptide-K, blocked Hsp90/Hsc70-dependent transformation of the heme-regulated eIF2 alpha kinase (HRI) into an active, heme-regulatable kinase. In contrast, peptide-K, but not peptide-Phi, inhibited Hsc70-mediated suppression of the activation of mature-transformed HRI. Furthermore, HDJ2 (Human DnaJ homologue 2), but not HDJ1, potentiated the ability of Hsc70 to suppress the activation of HRI in RRL. Mechanistically, peptide-K inhibited, while peptide-Phi enhanced, HDJ2 induced stimulation of Hsc70 ATPase activity in vitro. The data presented support the hypotheses that peptide-Phi acts to inhibit Hsc70 function by binding to the hydrophobic peptide-binding cleft of Hsc70, while peptide-K acts through binding to a site that modulates the interaction of Hsc70 with DnaJ homologues. Overall, the data indicate that peptide-Phi and peptide-K have differential effects on Hsc70 functions under quasi-physiological conditions in RRL, and suggest that therapeutically valuable peptide mimetics can be designed to inhibit specific functions of Hsc70. PMID- 11888293 TI - Characterization of the 2-phospho-L-lactate transferase enzyme involved in coenzyme F(420) biosynthesis in Methanococcus jannaschii. AB - The protein product of the Methanococcus jannaschii MJ1256 gene has been expressed in Escherichia coli, purified to homogeneity, and shown to be involved in coenzyme F(420) biosynthesis. The protein catalyzes the transfer of the 2 phospholactate moiety from lactyl (2) diphospho-(5')guanosine (LPPG) to 7,8 didemethyl-8-hydroxy-5-deazariboflavin (Fo) with the formation of the L-lactyl phosphodiester of 7,8-didemethyl-8-hydroxy-5-deazariboflavin (F(420)-0) and GMP. On the basis of the reaction catalyzed, the enzyme is named LPPG:Fo 2-phospho-L lactate transferase. Since the reaction is the fourth step in the biosynthesis of coenzyme F(420), the enzyme has been designated as CofD, the product of the cofD gene. The transferase requires Mg(2+) for activity, and the catalysis does not appear to proceed via a covalent intermediate. To a lesser extent CofD also catalyzes a number of additional reactions that include the formation of Fo-P, when the enzyme is incubated with Fo and GDP, GTP, pyrophosphate, or tripolyphosphate, and the hydrolysis of F(420)-0 to Fo. All of these side reactions can be rationalized as occurring by a common mechanism. CofD has no recognized sequence similarity to any previously characterized enzyme. PMID- 11888294 TI - Role of the N-terminal hydrophilic domain of acyl-coenzyme A:cholesterol acyltransferase 1 on the enzyme's quaternary structure and catalytic efficiency. AB - Acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an enzyme involved in cellular cholesterol homeostasis and atherosclerosis. ACAT1 is an allosteric enzyme responding to its substrate cholesterol in a sigmoidal manner. It is a homotetrameric protein that spans the membrane multiple times, with its N terminal 131 hydrophilic amino acids residing at the cytoplasmic side of the endoplasmic reticulum. This region contains two closely linked putative alpha helices. Our current studies show that this region contains a dimer-forming motif. Adding this motif to the bacterial glutathione S-transferase (GST) converted the homodimeric GST to a tetrameric fusion protein. Conversely, deleting this motif from the full-length ACAT1 converted the enzyme from a homotetramer to a homodimer. The dimeric ACAT1 remains enzymatically active. Its biochemical characteristics, including the sigmoidal response to cholesterol, the IC(50) value toward a specific ACAT inhibitor, and sensitivity toward heat inactivation, are essentially unaltered. On the other hand, the dimeric ACAT1 exhibits a 5-10-fold increase in the V(max) of the overall reaction and a 2.2 fold increase in the K(m) for oleoyl-coenzyme. Thus, deleting the dimer-forming motif near the N-terminus changes ACAT1 from its tetrameric form to a dimeric form and increases its catalytic efficiency. PMID- 11888295 TI - Reactivity, secondary structure, and molecular topology of the Escherichia coli sulfite reductase flavodoxin-like domain. AB - The flavodoxin-like domain, missing in the three-dimensional structure of the monomeric, simplified model of the Escherichia coli sulfite reductase flavoprotein component (SiR-FP), has now been expressed independently. This 168 amino acid protein was named SiR-FP18 with respect to its native molecular weight and represents the FMN-binding domain of SiR-FP. This simplified biological object has kept the main characteristics of its counterpart in the native protein. It could incorporate FMN exclusively and stabilize a neutral air-stable semiquinone radical. Both the radical and the fully reduced forms of SiR-FP18 were able to transfer their electrons to DCPIP or cytochrome c quantitatively. SiR-FP18 was able to form a highly stable complex with SiR-HP, the hemoprotein component of the sulfite reductase containing an iron-sulfur cluster coupled to a siroheme. In agreement with the postulated catalytic cycle of SiR-FP, only the fully reduced form of SiR-FP18 could transfer one electron to SiR-HP, the transferred electron being localized exclusively on the heme. As isolated SiR FP18 has kept the main characteristics of the FMN-binding domain of the native protein, a structural analysis by NMR was performed in order to complete the partial structure obtained previously. Structural modeling was performed using sequence homologues, cytochrome P450 reductase (CPR; 29% identity) and bacterial cytochrome P450 (P450-BM3; 26% identity), as conformational templates. These sequences were anchored using common secondary structural elements identified from heteronuclear NMR data measured on the protein backbone. The resulting structural model was validated, and subsequently refined using residual (C(alpha) C', N-H(N), and C'-H(N)) dipolar couplings measured in an anisotropic medium. The overall fold of SiR-FP18 is very similar to that of bacterial flavodoxins and of the flavodoxin-like domain in CPR or P450-BM3. PMID- 11888296 TI - Purification and characterization of the recombinant Na(+)-translocating NADH:quinone oxidoreductase from Vibrio cholerae. AB - The nqr operon from Vibrio cholerae, encoding the entire six-subunit, membrane associated, Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR), was cloned under the regulation of the P(BAD) promoter. The enzyme was successfully expressed in V. cholerae. To facilitate molecular genetics studies of this sodium pumping enzyme, a host strain of V. cholerae was constructed in which the genomic copy of the nqr operon was deleted. By using a vector containing a six-histidine tag on the carboxy terminus of the NqrF subunit, the last subunit in the operon, the recombinant enzyme was readily purified by affinity chromatography in a highly active form from detergent-solubilized membranes of V. cholerae. The recombinant enzyme has a high specific activity in the presence of sodium. NADH consumption was assessed at a turnover number of 720 electrons per second. When purified using dodecyl maltoside (DM), the isolated enzyme contains approximately one bound ubiquinone, whereas if the detergent LDAO is used instead, the quinone content of the isolated enzyme is negligible. Furthermore, the recombinant enzyme, purified with DM, has a relatively low rate of reaction with O(2) (10-20 s(-1)). In steady state turnover, the isolated, recombinant enzyme exhibits up to 5-fold stimulation by sodium and functions as a primary sodium pump, as reported previously for Na(+)()-NQR from other bacterial sources. When reconstituted into liposomes, the recombinant Na(+)-NQR generates a sodium gradient and a Delta Psi across the membrane. SDS-PAGE resolves all six subunits, two of which, NqrB and NqrC, contain covalently bound flavin. A redox titration of the enzyme, monitored by UV-visible spectroscopy, reveals three n = 2 redox centers and one n = 1 redox center, for which the presence of three flavins and a 2Fe-2S center can account. The V. cholerae Na(+)-NQR is well-suited for structural studies and for the use of molecular genetics techniques in addressing the mechanism by which NADH oxidation is coupled to the pumping of Na(+) across the membrane. PMID- 11888297 TI - Mutational analysis identifies a short atypical membrane attachment sequence (KYWFYR) within caveolin-1. AB - Caveolae are vesicular invaginations of the plasma membrane. Their formation is strictly dependent on the expression of the caveolin coat proteins. During transit to the plasma membrane, approximately 15 monomers of caveolin-1 assemble into a multivalent homo-oligomer. Caveolae are most likely generated through the subsequent interaction of these caveolin homo-oligomers with one another, with sphingolipids, and with cholesterol. Membrane association of caveolin-1 is critical to this process and is facilitated by an atypical N-terminal membrane attachment domain (residues 82-101), termed N-MAD. To better understand the membrane attachment function of N-MAD, we performed a detailed mutational analysis of the 20 amino acid N-MAD peptide sequence fused to the C-terminus of the soluble reporter green fluorescent protein (GFP). Removal of the distal six residues (KYWFYR) within N-MAD prevents membrane attachment in cells as assessed by hypotonic lysis, detergent solubility, carbonate extraction, and fluorescence microscopy. These six residues (KYWFYR) are sufficient to confer membrane attachment to GFP, an otherwise soluble protein. Both the central aromatic and flanking basic residues in this sequence are required for membrane attachment, as the sequence YWFY does not confer membrane affinity to GFP. Although the KYWFYR sequence within N-MAD facilitates membrane association, we show that the entire N MAD sequence is required for targeting to lipid rafts/caveolae. PMID- 11888298 TI - Phosphatidylglycerol requirement for the function of electron acceptor plastoquinone Q(B) in the photosystem II reaction center. AB - Phosphatidylglycerol (PG), a ubiquitous constituent of thylakoid membranes of chloroplasts and cyanobacteria, is demonstrated to be essential for the functionality of plastoquinone electron acceptor Q(B) in the photosystem II reaction center of oxygenic photosynthesis. Growth of the pgsA mutant cells of Synechocystis sp. PCC6803 that are defective in phosphatidylglycerolphosphate synthase and are incapable of synthesizing PG, in a medium without PG, resulted in a 90% decrease in PG content and a 50% loss of photosynthetic oxygen-evolving activity as reported [Hagio, M., Gombos, Z., Varkonyi, Z., Masamoto, K., Sato, N., Tsuzuki, M., and Wada, H. (2000) Plant Physiol. 124, 795-804]. We have studied each step of the electron transport in photosystem II of the pgsA mutant to clarify the functional site of PG. Accumulation of Q(A)(-) was indicated by the fast rise of chlorophyll fluorescence yield under continuous and flash illumination. Oxidation of Q(A)(-) by Q(B) plastoquinone was shown to become slow, and Q(A)(-) reoxidation required a few seconds when measured by double flash fluorescence measurements. Thermoluminescence measurements further indicated the accumulation of the S(2)Q(A)(-) state but not of the S(2)Q(B)(-) state following the PG deprivation. These results suggest that the function of Q(B) plastoquinone was inactivated by the PG deprivation. We assume that PG is an indispensable component of the photosystem II reaction center complex to maintain the structural integrity of the Q(B)-binding site. These findings provide the first clear identification of a specific functional site of PG in the photosynthetic reaction center. PMID- 11888299 TI - Interaction of internal water molecules with the schiff base in the L intermediate of the bacteriorhodopsin photocycle. AB - In the photocycle of bacteriorhodopsin (BR), the first proton movement, from the Schiff base to Asp85, occurs after the formation of the L intermediate. In L, the C [double bond] N bond of the Schiff base is strained, and the nitrogen interacts strongly with its counterion. The present study seeks to detect the interaction of internal water molecules with the Schiff base in L using difference FTIR spectroscopy at 170 K. The coupled modes of the hydrogen-out-of plane bending vibrations (HOOPs) of the N-H and C(15)-H of the protonated Schiff base are detected as a broad band centered at 911 cm(-1) for BR. A set of bands at 1073, 1064, and 1056 cm(-1) for L is shown to arise from the coupling of the HOOP with the overtones of interacting water O-H vibrations. Interaction with water was shown by the decreased intensity of the HOOPs of L in H(2)(18)O and by the influence of mutants that have been shown to perturb specific internal water molecules in BR. In contrast, the HOOP band of initial BR was not affected by these mutations. In D85N, the coupled HOOP of BR is depleted, while the coupled HOOPs of L are shifted. The results indicate that the Schiff base interacts with water in the L state but in a different manner than in the BR state. Moreover, the effects of mutations suggest that cytoplasmic water close to Thr46 (Wat46) either interacts stronger with the Schiff base in L or that it is important in stabilizing another water that does. PMID- 11888300 TI - Biochemical and functional characterization of recombinant Rhodnius prolixus platelet aggregation inhibitor 1 as a novel lipocalin with high affinity for adenosine diphosphate and other adenine nucleotides. AB - The Rhodnius prolixus aggregation inhibitor 1 (RPAI-1) is a novel blood-sucking salivary molecule that binds to ADP and attenuates platelet aggregation. In this report, we determine the binding constants and specificity of RPAI-1 for adenine nucleotides and its functional significance. By the Hummel-Dreyer method of equilibrium gel filtration, we show that RPAI-1 binds ADP with a K(0.5) of 48.6 plus minus 12.2 nM. RPAI-1 also displays high-affinity binding to ATP, AMP, Ado, AP4A, and alpha,beta Met ADP; however, RPAI-1 does not bind to inosine, guanosine, uridine, or cytidine. Binding is not modified by EDTA, indicating that Ado structure but not phosphate groups or Ca(2+) is necessary for binding. By computer simulation, we show that RPAI-1 is more effective in scavenging low but not high concentrations of ADP, in contrast to R. prolixus apyrase. RPAI-1 inhibits in vitro the ADP-dependent platelet-rich plasma aggregation by collagen (COLL), TRAP, PAF, and A23187 but did not block platelet aggregation by ristocetin or phorbol myristate acetate (PMA) and only slightly attenuated that by convulxin. RPAI-1 prolongs the closure time as assessed with PFA-100, when COLL-Epi but not COLL-ADP cartridges are employed. RPAI-1 also affects platelet mediated hemostasis time and COLL-induced thrombus formation at high shear as assessed with the Clot Signature Analyzer. We conclude that RPAI-1 exerts an antiplatelet effect due to scavenging of low concentrations of ADP in vitro and in vivo. RPAI-1 is the first lipocalin described so far with unique specificity for adenine nucleotides. PMID- 11888301 TI - Guanidinium chloride- and urea-induced unfolding of the dimeric enzyme glucose oxidase. AB - We have carried out a systematic study on the guanidinium chloride- and urea induced unfolding of glucose oxidase from Aspergillus niger, an acidic dimeric enzyme, using various optical spectroscopic techniques, enzymatic activity measurements, glutaraldehyde cross-linking, and differential scanning calorimetry. The urea-induced unfolding of GOD was a two-state process with dissociation and unfolding of the native dimeric enzyme molecule occurring in a single step. On the contrary, the GdmCl-induced unfolding of GOD was a multiphasic process with stabilization of a conformation more compact than the native enzyme at low GdmCl concentrations and dissociation along with unfolding of enzyme at higher concentrations of GdmCl. The GdmCl-stabilized compact dimeric intermediate of GOD showed an enhanced stability against thermal and urea denaturation as compared to the native GOD dimer. Comparative studies on GOD using GdmCl and NaCl demonstrated that binding of the Gdm(+) cation to the enzyme results in stabilization of the compact dimeric intermediate of the enzyme at low GdmCl concentrations. An interesting observation was that a slight difference in the concentration of urea and GdmCl associated with the unfolding of GOD was observed, which is in violation of the 2-fold rule for urea and GdmCl denaturation of proteins. This is the first report where violation of the 2-fold rule has been observed for a multimeric protein. PMID- 11888302 TI - Role of magnesium in nucleotide exchange on the small G protein rac investigated using novel fluorescent Guanine nucleotide analogues. AB - Novel guanine nucleotide analogues have been used to investigate the role of Mg(2+) in nucleotide release and binding with the small G protein rac. The fluorescent analogues have 7-(ethylamino)-8-bromocoumarin-3-carboxylic acid attached to the 3'-position of the ribose via an ethylenediamine linker. This modification has only small effects on the interaction with rac. There are large fluorescence changes on binding of the triphosphate to rac, on hydrolysis, and then on release of the diphosphate. Furthermore, the fluorescence is sensitive to the presence of Mg(2+) in the active site. Using this signal, it was shown that, for a variety of conditions, the nucleotides dissociate by a two-step mechanism. Mg(2+) is released first followed by the nucleotide. With the diphosphate, Mg(2+) is fast and nucleotide release slow. For the fluorescent GMPPNP analogue, the rate of dissociation is limited by Mg(2+) release. In the latter case, Mg(2+) binds tightly with a K(d) of 61 nM, whereas for the diphosphate the K(d) is 11 microM (30 degrees C, pH 7.6). PMID- 11888303 TI - Modulation of the internal aldimine pK(a)'s of 1-aminocyclopropane-1-carboxylate synthase and aspartate aminotransferase by specific active site residues. AB - The active sites of the homologous pyridoxal phosphate- (PLP-) dependent enzymes 1-aminocyclopropane-1-carboxylate (ACC) synthase and aspartate aminotransferase (AATase) are almost entirely conserved, yet the pK(a)'s of the two internal aldimines are 9.3 and 7.0, respectively, to complement the substrate pK(a)'s (S adenosylmethionine pK(a) = 7.8 and aspartate pK(a) = 9.9). This complementation is required for maximum enzymatic activity in the physiological pH range. The most prominent structural difference in the active site is that Ile232 of ACC synthase is replaced by alanine in AATase. The I232A mutation was introduced into ACC synthase with a resulting 1.1 unit decrease (from 9.3 to 8.2) in the aldimine pK(a), thus identifying Ile232 as a major determinant of the high pK(a) of ACC synthase. The mutation also resulted in reduced k(cat) (0.5 vs 11 s(-1)) and k(cat)/K(m) values (5.0 x 10(4) vs 1.2 x 10(6) M(-1) s(-1)). The effect of the mutation is interpreted as the result of shortening of the Tyr233-PLP hydrogen bond. Addition of the Y233F mutation to the I232A ACC synthase to generate the double mutant I232A/Y233F raised the pK(a) from 8.2 to 8.8, because the Y233F mutation eliminates the hydrogen bond between that residue and PLP. The introduction of the retro mutation A224I into AATase raised the aldimine pK(a) of that enzyme from 6.96 to 7.16 and resulted in a decrease in single-turnover k(max) (108 vs 900 s(-1) for aspartate) and k(max)/K(m)(app) (7.5 x 10(4) vs 3.8 x 10(5) M(-1) s(-1)) values. The distance from the pyridine nitrogen of the cofactor to a conserved aspartate residue is 2.6 A in AATase and 3.8 A in ACC synthase. The D230E mutation introduced into ACC synthase to close this distance increases the aldimine pK(a) from 9.3 to 10.0, as would be predicted from a shortened hydrogen bond. PMID- 11888304 TI - Lipase-catalyzed synthesis of a pure macrocyclic polyester from dimethyl terephthalate and diethylene glycol. AB - The polymerization of dimethyl terephthalate and diethylene glycol by enzymic catalysis in toluene is described. The potential pi-pi interactions of the aromatic rings together with the relative flexibility of the diol segment have been regarded as synergy factors responsible for the formation of a pure cyclic compound. The so-called C2 macrocycle was characterized by (1)H and (13)C NMR, size-exclusion chromatography, differential scanning calorimetry, mass spectrometry, and X-ray diffraction on powder. Structurally speaking, it is made of two repeating units. The substitution of the central oxygen atom of the diol by -CH(2)- or -S- as well as the presence of ortho-substituents (-NH(2) or NO(2)) on the phthalate were expected to disrupt the stacking of the phenyl groups more or less. PMID- 11888305 TI - Quadrupole time-of-flight mass spectrometry of the native hemocyanin of the deep sea crab Bythograea thermydron. AB - Since electrospray ionization mass spectrometry (ESI-MS) has demonstrated capabilities for observing intact, weak interactions, there has been increasing interest in studying by this method noncovalently bound complexes. In this communication, we report for the first time the structure obtained by a commercial ESI quadrupole time-of-flight spectrometer on a native hemocyanin of deep-sea crab Bythograea thermydron with a molecular mass of 1.3 MDa. ESI-MS analysis of the native hemocyanin revealed the formation of a 18-mer noncovalent assembly with a measured molecular mass of 1354940 +/- 480 Da. ESI-MS data also revealed that this huge structure is an equilibrium with several assemblages, dodecamer (measured molecular weight = 902570 +/- 110 Da), hexamer (measured molecular weight = 450310 +/- 260 Da), and monomeric structures (measured molecular weight = 74999 +/- 85 Da). PMID- 11888306 TI - Electrospinning of collagen nanofibers. AB - Electrospinning is a fabrication process that uses an electric field to control the deposition of polymer fibers onto a target substrate. This electrostatic processing strategy can be used to fabricate fibrous polymer mats composed of fiber diameters ranging from several microns down to 100 nm or less. In this study, we describe how electrospinning can be adapted to produce tissue engineering scaffolds composed of collagen nanofibers. Optimizing conditions for calfskin type I collagen produced a matrix composed of 100 nm fibers that exhibited the 67 nm banding pattern that is characteristic of native collagen. The structural properties of electrospun collagen varied with the tissue of origin (type I from skin vs type I from placenta), the isotype (type I vs type III), and the concentration of the collagen solution used to spin the fibers. Electrospinning is a rapid and efficient process that can be used to selectively deposit polymers in a random fashion or along a predetermined and defined axis. Toward that end, our experiments demonstrate that it is possible to tailor subtle mechanical properties into a matrix by controlling fiber orientation. The inherent properties of the electrospinning process make it possible to fabricate complex, and seamless, three-dimensional shapes. Electrospun collagen promotes cell growth and the penetration of cells into the engineered matrix. The structural, material, and biological properties of electrospun collagen suggest that this material may represent a nearly ideal tissue engineering scaffold. PMID- 11888307 TI - Elaboration and characterization of whey protein beads by an emulsification/cold gelation process: application for the protection of retinol. AB - Whey protein beads were successfully produced using a new emulsification/cold gelation method. The principle of this method is based on an emulsifying step followed by a Ca(2+)-induced gelation of pre-denatured (80 degreesC/30 min) whey protein. Beads are formed by the dropwise addition of the suspension into a calcium chloride (CaCl(2)) solution. IR results show that bead formation has a pronounced effect on the secondary structure of whey protein, which leads to the formation of intermolecular hydrogen-bonded beta-sheet structures. Their preparation conditions (CaCl(2) concentrations of 10, 15, and 20% (w/w)) influence their sphericity and homogeneity: an increase in CaCl(2) favors regular shaped beads. The physicochemical and mechanical characterizations of beads were also carried out. Their properties, such as swelling, elasticity, deformability, and resistance at fracture, change according to pH levels (1.9, 4.5, and 7.5) and preparation conditions. Indeed, protein chain networks exhibit different behavior patterns with respect to their charge. Finally, bead degradation by enzymatic hydrolysis reveals that beads are gastroresistant and form good matrixes to protect fat-soluble bioactive molecules such as retinol, that have in vivo intestinal absorption sites. The experiment demonstrated the potential of whey protein beads to protect molecules sensitive (i.e., vitamins) to oxidation. PMID- 11888308 TI - Photocurable liquid biodegradable copolymers: in vitro hydrolytic degradation behaviors of photocured films of coumarin-endcapped poly(epsilon-caprolactone-co trimethylene carbonate). AB - Coumarin-endcapped tetrabranched liquid copolymers composed of epsilon caprolactone and trimethylene carbonate (TMC), prepared using pentaerythritol or four-branched poly(ethylene glycol) (PEG) as an initiator, were ultraviolet irradiated to produce photocured solid biodegradable copolymers. The hydrolytic degradation behaviors of photocured films were determined from the weight loss of the films. The initial hydrolysis rate (determined for up to 24 h using a quartz crystal microbalance) was enhanced with aqueous solutions of higher pH. The hydrolysis rate in the early period of immersion was increased with an increase in TMC content, whereas that in the later period (week order) decreased with a increase in TMC content. This inverse relation of composition dependence on the hydrolysis rate between the early and late periods was discussed. Topological measurements using scanning electron microscopy and atomic force microscopy as well as depth profiles of the fluorescein-stained hydrolyzed layer showed that for the pentaerythritol-initiated copolymer, irrespective of copolymer composition, hydrolysis occurred at surface regions and surface erosion proceeded with immersion time. For PEG-based copolymers, both surface erosion and bulk degradation occurred simultaneously. The hydrolyzed surfaces became highly wettable with water and exhibited noncell adhesivity. PMID- 11888309 TI - The role of phasins in the morphogenesis of poly(3-hydroxybutyrate) granules. AB - Recent developments in the understanding of the structure of polyhydroxyalkanoate, PHA, granules in bacteria are documented in the literature and point to the role of structural proteins, phasins, in granule formation and stabilization. We have previously conceived a computer program which successfully simulates granule formation in vitro, in the absence of phasins. Now we are extending the computer model to a more complex system, including phasins, to quantify their anticipated effect on the granule properties. The simulation enabled us to propose real experiments to test the validity of the model and provide a framework for a better understanding of PHA granule formation in vivo. PMID- 11888311 TI - A new strategy for the preparation of supramolecular neutral hydrogels. AB - This paper demonstrates that miscible blends from water-insoluble polymers, such as poly(2,4,4-trimethylhexamethylene terephthalamide) (1), methylamine imidized poly(methyl methacrylate) (2), and aromatic poly(ether sulfone) (3) and water soluble polymers, such as poly(2-ethyl-2-oxazoline) (4) and poly(N-vinyl pyrrolidone) (5), respectively, represent a new class of supramolecular hydrogels. When the degree of polymerization (DP) of the water-soluble polymer is larger than that of water-insoluble polymer, the resulting hydrogels adsorb extremely high amounts of water (i.e., 229 wt % in the case of the hydrogel 1/4) and remain mechanically tough. The high water uptake capability of these blends is explained by a supramolecular network structure generated by H-bonding and/or other noncovalent interactions between the water-insoluble hydrophobic polymer and water-soluble hydrophilic segments as reversible cross-linking points interconnected by hydrophilic water soluble segments. The glass transition temperatures of these hydrogels are tailored via the ratio between the weight percent of the two polymers and by the glass transition temperature of the parent polymers. These supramolecular hydrogels can be processed from melt or solution and maintain excellent mechanical properties both in dry and in the water swollen state. This class of hydrogels is of interest for areas such as membranes, contact lenses, tissue engineering, and other biomedical applications. PMID- 11888310 TI - A tunable hydrogel for encapsulation and controlled release of bioactive proteins. AB - A N,N-dimethylacrylamide-based hydrogel (2) with the new cross-linker (ethylenedioxy) bis[2,2'-(N-acryloylamino)ethane] (1) has been prepared, and its physicochemical properties in aqueous solution were studied. Three different native proteins (lysozyme, bovine serum albumin, and rabbit IgG) were encapsulated within the polymeric matrix 2, and the kinetics of their release from the swollen hydrogel were determined. The rate of protein release exhibits a clear dependence on both the molecular weight of the protein and the amount of cross-linker utilized to prepare the hydrogel. This is reflected by the fact that the low molecular weight proteins are released at an increased rate versus higher molecular weight proteins. In addition a greater amount of protein is released from the hydrogels with a lower percentage of cross-linker. The polymerization procedure used in this study is sufficiently mild to safeguard the functional integrity of attendant biomolecules as determined by the retention of catalytic activity of encapsulated alpha-chymotrypsin and aldolase catalytic antibody 38C2. The potential utility of these hydrogels for the controlled release of bioactive agents in vivo is strengthened by both their lack of toxicity against human dermal fibroblasts and their lack of immunogenicity in mice. PMID- 11888312 TI - Amphiphilic block copolymer with a molecular recognition site: induction of a novel binding characteristic of amylose by self-assembly of poly(ethylene oxide) block-amylose in chloroform. AB - Solution properties of amphiphilic methoxy poly(ethylene oxide)-block-amyloses (MPEO-amyloses) in chloroform were investigated by SLS and DLS. The results indicated that MPEO-amyloses dissolved in chloroform containing 2 wt % DMSO by their self-associations. The complexation of MPEO-amylose with methyl orange (MO) was significantly enhanced in the amylose domain of the associate in chloroform. The blue shift of the maximum absorption and strong induced circular dichroism with exciton coupling were observed in the MPEO-amylose MO complex in chloroform. The self-assembly of MPEO-amylose in chloroform shows a unique feature for binding with MO. MPEO-block-amylose is a novel amphiphilic polymer with amylose as a molecular recognition site. PMID- 11888313 TI - Enzymatic degradation of and bovine serum albumin release from starch-acetate films. AB - The effect of acetylation of potato starch on swelling, enzymatic degradation, and bovine serum albumin (BSA, molecular mass 68 kDa) release rate from polymer films was studied. Potato starch and potato starch acetates (SA), having a degree of substitution of 1.9 or 2.6, were investigated. Polymer films were incubated in phosphate buffer solution pH 7.4 in the absence and presence of enzymes (alpha amylase, amyloglucosidase, esterase) or in human serum. The acetylation of potato starch decreased its swelling considerably. Increased acetylation of starch also considerably retarded its enzymatic degradation. Due to the decreased swelling and degradation of SA films, BSA was released much slower from SA films than from potato starch films, both in the presence and absence of enzymes. PMID- 11888314 TI - Characterization of an extracellular medium-chain-length poly(3-hydroxyalkanoate) depolymerase from Pseudomonas alcaligenes LB19. AB - An extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase from an isolate, Pseudomonas alcaligenes LB19, was purified to electrophoretic homogeneity by hydrophobic interaction chromatography using Octyl Sepharose CL-4B and gel permeation chromatography using Sephadex G-150. The molecular mass of the enzyme, which consisted of a single polypeptide chain, was approximately 27.6 kDa. The pI value of the enzyme was estimated to be 5.7, and its maximum activity was observed at pH 9.0 and 45 degreesC. The enzyme was significantly inactivated by EDTA and phenylmethylsulfonyl fluoride (PMSF) but insensitive to dithiothreitol. It was also markedly inhibited by 0.1% Tween 80 and 0.05% Triton X-100. The purified enzyme could hydrolyze various types of bacterial aliphatic and aromatic MCL-PHAs but not poly(3-hydroxybutyrate), polycaprolactone, and poly(L-lactide). Biodegradation rates of the aromatic MCL PHAs were significantly lower than those of the aliphatic MCL-PHAs, regardless of the compositions and types of aromatic substituents. It was able to hydrolyze medium-chain-length p-nitrophenylalkanoates more efficiently than the shorter chain forms. The main hydrolysis products of poly(3-hydroxynonanoate) were identified as monomer units. The results demonstrated in this study suggest that the MCL-PHA depolymerase from P. alcaligenes LB19 is a distinct enzyme, which are different from those of other MCL-PHA degrading bacteria in its quaternary structure, pI value, sensitivity to EDTA and PMSF, and hydrolysis products of MCL PHA. PMID- 11888316 TI - Modulated release of a volatile compound from starch matrixes via enzymatically controlled degradation. AB - The release of a model volatile (diacetyl) from a system based on a starch matrix, in which the volatile is dispersed, was studied. Kneading was used to obtain a homogeneous mixture (melt) composed of starch, glycerol alpha-amylase, and diacetyl. Samples were then ground to powders. When the starch powders were exposed to 30% relative humidity (RH) at 20 degreesC, no degradation of the starch matrix occurred. The samples only showed an initial burst release of diacetyl (around 10% of the loaded dose), whereas the remaining amount of diacetyl was not released, most likely due to the glassy character of the matrix and the low solubility of diacetyl in the matrix. However, when the samples were incubated at 90% RH, due to the uptake of moisture by the particles full release of the entrapped volatile occurred. The release of diacetyl from the matrix without enzyme followed first-order kinetics and, as expected, the release rate increased with decreasing particle size. Due to absorption of water, the enzyme became active and starch degradation occurred. The initial release of diacetyl from amylase-containing matrixes followed first-order kinetics as well. However, once the matrix was degraded to a certain extent, the particles collapsed, which was associated with concomitant rapid increase in release. The time at which the particle collapse occurred decreased with increasing enzyme concentration in the matrix. In conclusion, it is demonstrated that the release of a volatile from starch matrixes can be modulated both by the amount of coencapsulated matrix degrading enzyme and by the humidity of the environment. PMID- 11888315 TI - Elastin-based biopolymers: chemical synthesis and structural characterization of linear and cross-linked poly(OrnGlyGlyOrnGly). AB - Poly(OrnGlyGlyOrnGly) was synthesized by classical procedures in solution. The monomeric sequence -OrnGlyGlyOrnGly- was chosen as a modification of ValGlyGlyValGly-, typical of elastin, to impart primary amine functionality, susceptible to cross-linking with appropriate bifunctional reagents. Herein we focus on the cross-linking of poly(OrnGlyGlyOrnGly) with glutaraldehyde. The polymers, both linear and cross-linked, were characterized and investigated for their molecular and supramolecular properties. Circular dichroism studies performed on linear poly(OrnGlyGlyOrnGly) revealed a variety of conformations similar to elastin. At a supramolecular level, different kinds of aggregates were found such as the elastin-like twisted-rope pattern of filaments and fibrils, together with other specific morphologies, similar to those recently identified in some elastin-mimetic polypeptides. PMID- 11888317 TI - Nonhydrolytic fragmentation of a poly[(R)-3-hydroxybutyrate] single crystal revealed by use of a mutant of polyhydroxybutyrate depolymerase. AB - This paper reports the initial process of the enzymatic degradation of solution grown lamellar single crystals of bacterial poly[(R)-3-hydroxybutyrate] (P(3HB)) with an extracellular polyhydroxybutyrate (PHB) depolymerase purified from Alcaligenes faecalis T1. We used a hydrolytic-activity-disrupted mutant of the PHB depolymerase in order to avoid the influence of hydrolytic reaction in the system. The effect of addition of the mutant enzyme upon the P(3HB) single crystals was investigated by turbidimetric assay, high-performance liquid chromatography (HPLC), and atomic force microscopy (AFM). Suspension turbidity of the P(3HB) single crystals increased after addition of the mutant enzyme having no hydrolytic activity. No soluble product from the P(3HB) single crystals with the mutant enzyme was detected by HPLC. AFM observation of the P(3HB) single crystals adsorbed on highly ordered pyrolytic graphite revealed that the mutant enzyme yielded a lot of lengthwise crystal fragments from the P(3HB) single crystals. On the basis of these results, we concluded that the mutant enzyme disturbs the molecular packing of the P(3HB) polymer chain around the loose chain packing region in the single crystal, resulting in the fragmentation. Therefore, it is suggested that the enzymatic degradation of P(3HB) single crystals with a wild-type PHB depolymerase progresses via three steps: (1) adsorption of the enzyme onto the surface of the single crystal; (2) disturbance of the molecular packing of P(3HB) polymer chain in the single crystal by the adsorbed enzyme; and (3) hydrolysis of the disturbed polymer chain by the adsorbed enzyme. PMID- 11888318 TI - Protease-catalyzed regioselective polymerization and copolymerization of glutamic acid diethyl ester. AB - Protease-catalyzed polymerization and copolymerization of L-glutamic acid diethyl ester hydrochloride (1) have been performed in a buffer of high concentration. Papain and bromelain showed high catalytic activity toward the polymerization. H H COSY NMR analysis of the product showed the exclusive formation of poly(alpha peptide), which was further confirmed by comparison with NMR spectra of poly(alpha-methyl gamma-L-glutamate). The papain-catalyzed polymerization of gamma-methyl L-glutamate did not occur under the similar reaction conditions, supporting the regioselective production of the polymer having an alpha-peptide linkage from 1. The effects of the reaction parameters have been systematically investigated. The copolymerization of 1 with various amino acid esters took place by the papain catalyst to give peptide copolymers. PMID- 11888319 TI - Mimicking a cytoskeleton by coupling poly(N-isopropylacrylamide) to the inner leaflet of liposomal membranes: effects of photopolymerization on vesicle shape and polymer architecture. AB - Networks of N-isopropylacrylamide (NIPAM) copolymers, coupled to spherical phospholipid bilayers, are suitable as a model for the study of the interaction between the cytoskeleton and cellular membranes, as well as for promising new drug delivery systems with triggerable drug release properties and improved stability. In this article, we describe a simple preparation technique for liposomes from egg phosphatidyl choline (EPC) encapsulating a cross-linked NIPAMminus signTEGDM copolymer skeleton (tetraethylene glycol dimethacrylate, TEGDM) which is coupled only to the inner monolayer by a novel membrane anchor monomer. Polymerization in the lipid vesicles was initiated at the inner membrane surface by the radical initiator 2,2-diethoxy-acetophenone (DEAP) permeating through the membrane from the outside. The effects of photopolymerization and polymer formation on vesicle shape and membrane integrity were studied by transmission electron microscopy (TEM), cryo-TEM, and atomic force microscopy (AFM). Upon UV irradiation, approximately 100% of the vesicles contained a polymer gel and only occasional changes in the spherical shape of the liposomes were observed. The architecture of the polymer network inside the liposomal compartment was determined by the conditions of the photopolymerization. Composite structures of polymer hollow spheres or solid spheres, respectively, tethered to spherical membrane vesicles were produced. The increased stability of the polymer-tethered lipid bilayers against solubilization by sodium cholate, compared to pure EPC vesicles, was determined by radiolabeling the lipid membrane. PMID- 11888320 TI - Enzymatic synthesis of inulin-containing hydrogels. AB - The Bacillus subtilis protease Proleather FG-F catalyzed the transesterification of inulin with vinyl acrylate (VA) in dimethylformamide (DMF). The reaction conversion for different VA concentrations was greater than 57% after 96 h at 50 degrees C. The degree of substitution (DS, defined as the amount of acrylate groups per 100 inulin fructofuranoside residues) with acrylate moieties can be controlled by varying the molar ratio of VA to inulin. Reasonable yields were obtained (44-51%, 2 days) using a two-step purification methodology. Inulin derivatized with VA (Inul-VA) was characterized by gel permeation chromatography, and its structure was established by (1)H, (13)C, and (1)H-(1)H correlation spectroscopy and (1)H-(13)C heteronuclear multiple quantum coherence NMR. The main positional isomer was at the 6 position of the fructofuranoside residue and two other minor isomers were observed at the 3 and 4 positions. Thus, the enzymatic reaction was largely regioselective. Furthermore, the inulin fructose residues were monosubstituted. Gels with swelling ratios at equilibrium of up to ca. 20 were prepared by free radical polymerization of aqueous solutions of Inul VA with different DS and monomer concentrations. Gel pore sizes were calculated from swelling experiments and range from 19 to 57 A. To our knowledge, this work reports the first successful enzymatic modification of a polysaccharide solubilized in 100% DMF solution. PMID- 11888321 TI - Viscosity of semiflexible chitosan solutions: influence of concentration, temperature, and role of intermolecular interactions. AB - The influence of polymer concentration and temperature on the rheological behavior of chitosan solution was studied. The threshold concentrations for the different viscometric regimes were determined and the different power laws exponents were calculated and compared with those predicted from models. Different observations and the high values of these exponents within the high concentration region lead to consideration of the presence of intermolecular interactions as soon as the polymer concentration is larger than the overlap concentration. The activation energy was determined as a function of the polymer concentration, and its evolution was compared with theoretical predictions. A gel sol transition was demonstrated at high concentrations. PMID- 11888322 TI - Crystallization, stability, and enzymatic degradation of poly(L-lactide) thin film. AB - Poly(L-lactide) (PLLA) thin film with 100 nm thickness was crystallized at 160 degreesC for 20 min from the melt obtained at 220 degreesC. Hexagonal crystals with three types of growth (derivative growth lamellae, overgrowth multistacked lamellae, and undergrowth multistacked lamellae) were simultaneously observed by atomic force microscopy (AFM). These phenomena are due to the differences of the formative points of secondary crystal nuclei against the basal lamella. Enzymatic degradation of PLLA thin film revealed two types of amorphous regions. These regions were identified as the free amorphous region around the crystals and the restricted amorphous region between the crystal and glass substrate. In situ observation of thermal behavior of lamellar crystals was performed to understand the correlation between the chain folding and stability of the crystal by using temperature-controlled AFM. The morphology of the sectors with [100] growth plane had changed to a comblike morphology despite the fact that the [110] growth plane remained unchanged, suggesting that the stability of the chain folding and the chain-packing state affected the thermal behavior. PMID- 11888323 TI - Genetically encoded synthesis of protein-based polymers with precisely specified molecular weight and sequence by recursive directional ligation: examples from the elastin-like polypeptide system. AB - We report a new strategy for the synthesis of genes encoding repetitive, protein based polymers of specified sequence, chain length, and architecture. In this stepwise approach, which we term "recursive directional ligation" (RDL), short gene segments are seamlessly combined in tandem using recombinant DNA techniques. The resulting larger genes can then be recursively combined until a gene of a desired length is obtained. This approach is modular and can be used to combine genes encoding different polypeptide sequences. We used this method to synthesize three different libraries of elastin-like polypeptides (ELPs); each library encodes a unique ELP sequence with systematically varied molecular weights. We also combined two of these sequences to produce a block copolymer. Because the thermal properties of ELPs depend on their sequence and chain length, the synthesis of these polypeptides provides an example of the importance of precise control over these parameters that is afforded by RDL. PMID- 11888324 TI - New routes to the synthesis of amylose-block-polystyrene rod-coil block copolymers. AB - Hybrid block copolymers amylose-block-polystyrene were synthesized by covalent attachment of maltoheptaose derivatives to end-functionalized polystyrene and subsequent enzymatic grafting from polymerization. The maltoheptaose derivatives were attached by reductive amination or hydrosilation to amino- or SiH-terminated polystyrene (synthesized by anionic polymerization), respectively. The enzymatic polymerization could be started even though the primer-modified polystyrenes were insoluble in the medium of polymerization, i.e., citrate buffer. The polymerization kinetics show an interesting dependence on the molecular weight of polystyrene due to the micellar structure of the primer in water. PMID- 11888325 TI - Synthesis of in situ cross-linkable macroporous biodegradable poly(propylene fumarate-co-ethylene glycol) hydrogels. AB - This study describes a synthesis method of biodegradable macroporous hydrogels suitable as in situ cross-linkable biomaterials. Macroporous hydrogels were based on poly(propylene fumarate-co-ethylene glycol) and prepared via coupled free radical and pore formation reactions. Cross-linking was initiated by a pair of redox initiators, ammonium persulfate and L-ascorbic acid. Pores were formed by the reaction between L-ascorbic acid and sodium bicarbonate, a basic component, which evolved carbon dioxide. Sol fraction of the hydrogels was varied from 0.06 +/- 0.01 to 0.64 +/- 0.01. A stereological approach was used to analyze the morphological properties of the macroporous hydrogels by relating the morphological properties of thin sections to the original three-dimensional macroporous hydrogel. Prepared macroporous hydrogels had porosities between 0.43 +/- 0.08 and 0.84 +/- 0.02 and surface area densities between 55 +/- 3 and 108 +/ 7 cm(-1). Sodium bicarbonate concentration had the greatest effect on both the porosity and surface area density. The effect of copolymer formulation on the porosity and surface area density was insignificant. From thin sections of the macroporous hydrogels, the profile size distributions were determined as an estimate of the pore size distribution. Two formulations synthesized with varying L-ascorbic acid concentration of 0.05 and 0.1 M had median profile sizes of 50 100 and 150-200 microm, respectively. This novel synthesis method allows for the in situ cross-linking of biodegradable macroporous hydrogels with morpholological properties suitable for consideration as an injectable tissue engineering scaffold. PMID- 11888326 TI - Fine-stranded and particulate aggregates of heat-denatured whey proteins visualized by atomic force microscopy. AB - beta-Lactoglobulin and whey protein isolate (WPI) were heated in aqueous solutions at pH 2 and 7 at 80 degrees C, spread onto freshly cleaved mica surfaces, and visualized under butanol using atomic force microscopy. Fine stranded aggregates were formed at pH 2, the diameter of strands being ca. 4 nm for beta-lactoglobulin and 10 nm for WPI. At pH 7, aggregates were composed of ellipsoidal particles, regardless of the concentration of added NaCl. This observation supports the previously proposed two-step aggregation model at neutral pH (Aymard, P.; Gimel, J. C.; Nicolai, T.; Durand, D. J. Chim. Phys. 1996, 93, 987-997), consisting of the formation of primary globular particles and the subsequent aggregation of those primary particles. The AFM provides the first direct evidence for the anisotropic shape of these primary particles. The heights of primary particles increased from ca. 11 to 27 nm with increasing concentrations of added NaCl from 0 to 0.3 M in the case of WPI. The rate of aggregation was also accelerated with increasing NaCl concentrations, which appeared to induce transitions in gel networks from fine-stranded toward particulate networks. The present study provides structural information essential for understanding the diverse physical properties of heat-induced whey protein gels. PMID- 11888327 TI - Crystalline/amorphous phase structure and molecular mobility of biodegradable poly(butylene adipate-co-butylene terephthalate) and related polyesters. AB - Differential scanning calorimetry (DSC), atomic force microscopy (AFM), wide angle X-ray scattering (WAXD), and solid-state (13)C NMR have been used to investigate the crystalline/amorphous structure and molecular mobility of biodegradable poly(butylene adipate-co-44 mol % butylene terephthalate) [P(BA-co 44 mol % BT)] copolyester sample crystallized from the melt. The DSC endothermic peak, which is ascribed to the melting of the crystalline region, was broad relative to those reported for conventional partially crystalline polyesters. In AFM observation, spherulitic morphology was not observed while small particles with a size of about 100 nm were detected. The WAXD pattern of the sample was very broad. These results have indicated that a melt-crystallized P(BA-co-44 mol % BT) sample contains small crystals with a wide distribution in size. A solid state (13)C NMR technique was also used to perform molecular-level and selective analyses for both butylene terephthalate and butylene adipate units. For the butylene terephthalate units, the existence of two components with different microstructure and molecular mobility was detected: one component was assigned to the alpha-form crystal of poly(butylene terephthalate) homopolymer (PBT) and the other was in amorphous regions. In contrast, all of butylene adipate units were located in amorphous regions. Solid-state NMR data have suggested that sizes of crystalline regions are less than 3 nm. PMID- 11888328 TI - Synthesis and characterization of self-assembling block copolymers containing bioadhesive end groups. AB - 3,4-Dihydroxyphenyl-L-alanine (DOPA) is an unusual amino acid found in mussel adhesive proteins (MAPs) that is believed to lend adhesive characteristics to these proteins. In this paper, we describe a route for the conjugation of DOPA moieties to poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO PPO-PEO) block copolymers. Hydroxyl end groups of PEO-PPO-PEO block copolymers were activated by N,N'-disuccinimidyl carbonate and then reacted with DOPA or its methyl ester with high coupling efficiencies from both aqueous and organic solvents. DOPA-modified PEO-PPO-PEO block copolymers were freely soluble in cold water, and dye partitioning and differential scanning calorimetry analysis of these solutions revealed that the copolymers aggregated into micelles at a characteristic temperature that was dependent on block copolymer composition and concentration in solution. Oscillatory rheometry demonstrated that above a block copolymer concentration of approximately 20 wt %, solutions of DOPA-modified PEO PPO-PEO block copolymers exhibited sol-gel transitions upon heating. The gelation temperature could be tailored between approximately 23 and 46 degrees C by changing the composition, concentration, and molecular weight of the block copolymer. Rheological measurement of the bioadhesive interaction between DOPA modified Pluronic and bovine submaxillary mucin indicated that DOPA-modified Pluronic was significantly more bioadhesive than unmodified Pluronic. PMID- 11888330 TI - Cellular and clinical pharmacology of fludarabine. AB - In the past decade, fludarabine has had a major impact in increasing the effectiveness of treatment of patients with indolent B-cell malignancies. This has come about in a variety of clinical circumstances, including use of fludarabine alone as well as in combinations with DNA-damaging agents or membrane targeted antibodies. Other strategies have used fludarabine to reduce immunological function, thus facilitating non-myeloablative stem cell transplants. Fludarabine is a prodrug that is converted to the free nucleoside 9 beta-D-arabinosyl-2-fluoroadenine (F-ara-A) which enters cells and accumulates mainly as the 5'-triphosphate, F-ara-ATP. The rate-limiting step in the formation of triphosphate is conversion of F-ara-A to its monophosphate, which is catalyzed by deoxycytidine kinase. Although F-ara-A is not a good substrate for this enzyme, the high specific activity of this protein results in efficient phosphorylation of F-ara-A in certain tissues. F-ara-ATP has multiple mechanisms of action, which are mostly directed toward DNA. These include inhibition of ribonucleotide reductase, incorporation into DNA resulting in repression of further DNA polymerisation, and inhibition of DNA ligase and DNA primase. Collectively these actions affect DNA synthesis, which is the major mechanism of F-ara-A-induced cytotoxicity. Secondarily, incorporation into RNA and inhibition of transcription has been shown in cell lines. With the standard dose of fludarabine (25 to 30 mg/m(2)/day given over 30 minutes for 5 days), plasma concentrations of about 3 micromol/L F-ara-A are achieved at the end of each infusion. Serial sampling of leukaemia cells from patients receiving these standard doses of fludarabine has demonstrated that the peak concentrations of F ara-ATP are achieved 4 hours after start of fludarabine infusion. Although there is heterogeneity among individuals with respect to rate of F-ara-ATP accumulation, the peak concentrations are generally proportional to the dose of the drug. Knowledge of the plasma pharmacokinetics of its principal nucleoside metabolite F-ara-A, and the cellular pharmacology of the proximal active metabolite, F-ara-ATP, has provided some understanding of the activity of fludarabine when used as a single agent. Preclinical studies directed toward learning the mechanisms of action of this agent have formed the basis for several mechanism-based strategies for its combination and scheduling with other agents. As a single agent fludarabine has been effective for the indolent leukaemias. Biochemical modulation strategies resulted in enhanced accumulation of cytarabine triphosphate and led to the use of fludarabine for the treatment of acute leukaemias. Combination of fludarabine with DNA damaging agents to inhibit DNA repair processes has been highly effective for indolent leukaemias and lymphomas. The current review brings together knowledge of the mechanisms of fludarabine, the state of understanding of the plasma pharmacokinetics, and cellular pharmacodynamics of fludarabine nucleotides. This may be useful in the design of future therapeutic approaches. PMID- 11888331 TI - Pharmacokinetic profile of zafirlukast. AB - Zafirlukast is a cysteinyl leukotriene type 1 receptor antagonist that causes bronchodilation and has anti-inflammatory properties. Clinical efficacy has been demonstrated when using oral doses of 20 to 40 mg twice daily. The pharmacokinetics of zafirlukast are best described by a two-compartment model. Maximum plasma concentrations (Cmax) were achieved 3 hours after a single oral dose of 20 or 40 mg to healthy volunteers. The absolute bioavailability of zafirlukast is unknown. However, coadministration of zafirlukast with food reduces bioavailability by approximately 40%. The drug binds to plasma proteins (>99%), predominantly to albumin, and has a mean terminal elimination half-life of approximately 10 hours in both healthy volunteers and patients with asthma. Zafirlukast undergoes extensive hepatic metabolism. Hydroxylation by cytochrome P450 (CYP) 2C9 is the major biotransformation pathway. The metabolites of zafirlukast contribute little to its overall activity. Zafirlukast is mainly eliminated in the faeces, while urinary excretion accounts for <10% of an orally administered dose. Because of its primarily hepatic metabolism, the clearance of zafirlukast is reduced in patients with hepatic impairment. In patients with stable alcoholic cirrhosis, Cmax and area under the plasma concentration-time curve for zafirlukast were increased by 50 to 60% compared with healthy volunteers. Asymptomatic elevations of serum liver enzymes have been reported with high dosages of zafirlukast (80 mg twice daily), returning to normal after cessation of the drug. Inhibition of the CYP2C9 and CYP3A isoenzymes by zafirlukast has been reported in vitro. Zafirlukast interacts with warfarin and produces a clinically significant increase in the prothrombin time, but it does not alter the pharmacokinetics of terfenadine carboxylate, the active metabolite of terfenadine. Plasma concentrations of zafirlukast decreased when the drug was administered concomitantly with erythromycin, terfenadine or theophylline, and increased when it was coadministered with aspirin (acetylsalicylic acid). Theophylline metabolism is unchanged in most cases by zafirlukast, but there is a report of one patient with increased theophylline plasma concentrations when zafirlukast was coadministered. Recently, cases of Churg-Strauss syndrome have been described in patients with asthma receiving zafirlukast treatment. This occurrence in patients being withdrawn from corticosteroid therapy while receiving zafirlukast has been attributed to a previously undiagnosed presence of this syndrome in these patients. PMID- 11888329 TI - The impact of efflux transporters in the brain on the development of drugs for CNS disorders. AB - The development of drugs to treat disorders of the CNS requires consideration of achievable brain concentrations. Factors that influence the brain concentrations of drugs include the rate of transport into the brain across the blood-brain barrier (BBB), metabolic stability of the drug, and active transport out of the brain by efflux mechanisms. To date, three classes of transporter have been implicated in the efflux of drugs from the brain: multidrug resistance transporters, monocarboxylic acid transporters, and organic ion transporters. Each of the three classes comprises multiple transporters, each of which has multiple substrates, and the combined substrate profile of these transporters includes a large number of commonly used drugs. This system of transporters may therefore provide a mechanism through which the penetration of CNS-targeted drugs into the brain is effectively minimised. The action of these efflux transporters at the BBB may be reflected in the clinic as the minimal effectiveness of drugs targeted at CNS disorders, including HIV dementia, epilepsy, CNS-based pain, meningitis and brain cancers. Therefore, modulation of these efflux transporters by design of inhibitors and/or design of compounds that have minimal affinity for these transporters may well enhance the treatment of intractable CNS disorders. PMID- 11888334 TI - Population pharmacokinetics of haloperidol using routine clinical pharmacokinetic data in Japanese patients. AB - OBJECTIVE: To clarify the observed variability of haloperidol disposition in patients with psychiatric disorders. DESIGN: Retrospective population pharmacokinetic study. PARTICIPANTS: 218 Japanese patients aged 16 to 82 years who provided 391 serum haloperidol concentrations. METHODS: Routine clinical pharmacokinetic data gathered from patients receiving haloperidol were analysed to estimate population pharmacokinetic parameters with the nonlinear mixed effects model (NONMEM) computer program. RESULTS: The final pharmacokinetic model was CL = 42.4 * (TBW/60)(0.655) * 0.814(AGE> or = 55) * (DOSE/200)(0.236) * 1.32(ANTIEP) and Vd = 34.4 * TBW * 0.336( AGE> or = 65), where CL is total body clearance (L/h), Vd is apparent volume of distribution (L), TBW is total bodyweight (kg), DOSE is daily dosage (microg/kg/day), ANTIEP = 1 for concomitant administration of antiepileptic drugs (phenobarbital, phenytoin or carbamazepine) and 0 otherwise, AGE > or = 55 = 1 for patient aged 55 years or over and 0 otherwise, and AGE > or = 65 = 1 for patient aged 65 years or over and 0 otherwise. Concomitant administration of haloperidol and antiepileptic drugs resulted in a 32% increase in haloperidol clearance. Patients aged 55 years or over showed an 18.6% reduction in clearance, and elderly patients aged 65 years or over showed a 66.4% reduction in apparent volume of distribution. Inclusion of terms for the concomitant administration of haloperidol and antiparkinsonian drugs (amantadine, bromocriptine, biperiden, trihexyphenidyl or mazaticol) or cytochrome P450 (CYP) 2D6 substrates (levomepromazine, perphenazine, thioridazine, amitriptyline or clomipramine) did not significantly improve the estimate of haloperidol clearance. CONCLUSION: Application of the findings in this study to patient care may permit selection of an appropriate initial maintenance dosage to achieve target haloperidol serum concentrations, thus enabling the clinician to achieve the desired therapeutic effect. PMID- 11888333 TI - Population pharmacokinetic-pharmacodynamic modelling of angiotensin receptor blockade in healthy volunteers. AB - OBJECTIVE: To compare the pharmacokinetic and pharmacodynamic characteristics of angiotensin II receptor antagonists as a therapeutic class. DESIGN: Population pharmacokinetic-pharmacodynamic modelling study. METHODS: The data of 14 phase I studies with 10 different drugs were analysed. A common population pharmacokinetic model (two compartments, mixed zero- and first-order absorption, two metabolite compartments) was applied to the 2685 drug and 900 metabolite concentration measurements. A standard nonlinear mixed effect modelling approach was used to estimate the drug-specific parameters and their variabilities. Similarly, a pharmacodynamic model was applied to the 7360 effect measurements, i.e. the decrease of peak blood pressure response to intravenous angiotensin challenge recorded by finger photoplethysmography. The concentration of drug and metabolite in an effect compartment was assumed to translate into receptor blockade [maximum effect (Emax) model with first-order link]. RESULTS: A general pharmacokinetic-pharmacodynamic (PK-PD) model for angiotensin antagonism in healthy individuals was successfully built up for the 10 drugs studied. Representatives of this class share different pharmacokinetic and pharmacodynamic profiles. Their effects on blood pressure are dose-dependent, but the time course of the effect varies between the drugs. CONCLUSIONS: The characterisation of PK PD relationships for these drugs gives the opportunity to optimise therapeutic regimens and to suggest dosage adjustments in specific conditions. Such a model can be used to further refine the use of this class of drugs. PMID- 11888332 TI - Immunosuppressive therapy for paediatric transplant patients: pharmacokinetic considerations. AB - Immunosuppressive therapy in paediatric transplant recipients is changing as a consequence of the increasing number of available immunosuppressive agents. Generic and other new formulations are now emerging onto the market, clinical experience is growing, and it is expected that clinicians should tailor immunosuppressive protocols to individual patients by optimising dosages and drugs according to the maturation and clinical status of the child. Most information about the clinical pharmacokinetics of immunosuppressive drugs in paediatrics is centred on cyclosporin, tacrolimus and mycophenolate mofetil in renal and liver transplant recipients; data regarding other immunosuppressants and transplant types are limited. Although the clinical pharmacokinetics of these drugs in paediatric transplant recipients are still under investigation, it is evident that the pharmacokinetic parameters observed in adults may not be applicable to children, especially in younger age groups. In general, patients younger than 5 years old show higher clearance rates irrespective of the organ transplanted or drug used. Another important factor that frequently affects clearance in this patient population is the post-transplant time. In accordance with these findings, and in contrast with the usual under-dosage in children, the need for higher dosages in younger recipients and during the early post transplant period seems evident. To achieve the best compromise between prevention of rejection and toxicity, dosage individualisation is required and this can be achieved through therapeutic drug monitoring (TDM). This approach is particularly useful to ensure the cost-effective management of paediatric transplant recipients in whom the pharmacokinetic behaviour, target concentrations for clinical use and optimal dosage strategies of a particular drug may not yet be well defined. Although TDM may be a tool for improving immunosuppressive therapy, there is little information concerning its positive contribution to clinical events, including outcomes, for paediatric patients. Substantial information to support the use of TDM exists for cyclosporin and, to a lesser extent, for tacrolimus, but a diversity of options affects their implementation in the clinical setting. The role of TDM in therapy with mycophenolate mofetil and sirolimus has yet to be defined regarding both methods and clinical indications. Pharmacodynamic monitoring appears more suited to other immunosuppressants such as azathioprine, corticosteroids and monoclonal or polyclonal antibodies. If coupled with pharmacokinetic measurements, such monitoring would allow earlier and more precise optimisation of therapy. Very few population pharmacokinetic studies have been carried out in paediatric transplant patients. This type of study is needed so that techniques such as Bayesian forecasting can be applied to optimise immunosuppressive therapy in paediatric transplant patients. PMID- 11888335 TI - Angiotensin AT2 receptor ligands: do they have potential as future treatments for neurological disease? AB - In addition to the systemic renin-angiotensin system (RAS), a local RAS has been identified. Recent research has focused on this latter system and has investigated the effects of locally generated angiotensin II, especially in the kidney, heart and CNS. In the mammalian brain, all components of the RAS are present including angiotensin AT(1) and AT(2) receptor subtypes. While the AT(1) receptor is responsible for the classical effects of angiotensin II, it has been found that the AT(2) receptor displays totally different signalling mechanisms and this has revealed hitherto unknown functions of angiotensin II. AT(2) receptors are expressed at low density in many healthy adult tissues, but are up regulated in pathological circumstances, e.g. stroke or nerve lesion. Evidence has now emerged that the actions of angiotensin II that are exerted via the AT(2) receptor are directly opposed to those mediated by the AT(1 )receptor. For example, the AT(2) receptor has antiproliferative properties and therefore opposes the growth-promoting effect linked to AT(1) receptor stimulation. It has been reported that the AT(2) receptor regulates several functions of nerve cells, e.g. ionic fluxes, cell differentiation and axonal regeneration, but also modulates programmed cell death. It is possible that a more extensive knowledge of the AT(2) receptor could contribute to the understanding of the clinically beneficial effects of AT(1) receptor antagonists, as this treatment may unmask AT(2) receptor activity. This review presents selected aspects of advances in AT(2) receptor pharmacology, molecular biology and signal transduction with particular reference to possible novel therapeutic options for CNS diseases. PMID- 11888336 TI - Place of polytherapy in the early treatment of epilepsy. AB - Polytherapy with antiepileptic drugs is not popular mainly because it is thought to be associated with more adverse effects and to contribute relatively little in terms of efficacy compared with monotherapy. However, there are two reasons to question this assumption: certain combinations are more effective than others and, therefore, generalisations about the poor effectiveness of polytherapy cannot be made; and the total drug load, i.e. the total amount of drug exposure for a certain indication, is usually higher in polytherapy, which may explain the higher toxicity seen during such treatment. In this article, the available literature on the effectiveness of first-line monotherapy, alternative monotherapy and second-line polytherapy is reviewed. There is no conclusive evidence in favour for choosing either alternative monotherapy or polytherapy when first-line monotherapy fails. Therefore, a pragmatic approach is recommended until an evidence-based choice can be made. PMID- 11888337 TI - Drug-induced Pisa syndrome (pleurothotonus): epidemiology and management. AB - Long-term administration of antipsychotics occasionally produces persistent dystonia of the trunk, a disorder known as Pisa syndrome (or pleurothotonus). The development of Pisa syndrome is most commonly associated with prolonged treatment with antipsychotics; however, it has also been reported, although less frequently, in patients who are receiving other medications (such as cholinesterase inhibitors and antiemetics), in those not receiving medication (idiopathic Pisa syndrome) and in those with neurodegenerative disorders. Drug induced Pisa syndrome predominantly develops in females and in older patients with organic brain changes. It sometimes occurs after the addition of another antipsychotic to an established regimen of antipsychotics or insidiously arises in antipsychotic-treated patients for no apparent reason. The condition generally disappears after antipsychotic drugs are discontinued. Although a pharmacological therapy for drug-induced Pisa syndrome has not been established, we have reported that anticholinergic drugs are effective in about 40% of patients who have episodes of Pisa syndrome with the remaining patients responding to the withdrawal or reduction of daily doses of antipsychotic drugs. The characteristics of its development and prognosis indicate that drug-induced Pisa syndrome consists of two types of dystonia. Some patients develop clinical features of acute dystonia, whereas others develop symptoms similar to tardive dystonia. Like that of tardive dystonia, Pisa syndrome responds better than tardive dyskinesia to a relatively high daily dose of an anticholinergic. However, the significant improvement caused by the withdrawal of antipsychotic drugs in Pisa syndrome differentiates it from tardive dystonia. Thus, Pisa syndrome including these features is considered to be an atypical type of tardive dystonia. These clinical characteristics suggest that the underlying pathophysiology of drug-induced Pisa syndrome is complex. A dopaminergic cholinergic imbalance, or serotonergic or noradrenergic dysfunction, may be implicated. Asymmetric brain functions or neural transmission may also be considered as underlying mechanisms of the development of Pisa syndrome that is resistant to anticholinergic drugs. Idiopathic Pisa syndrome is characterised by an adult-onset, segmental truncal dystonia in patients with no previous exposure to antipsychotics. It occurs rarely but shows a complete resolution with high doses of anticholinergic drugs. PMID- 11888338 TI - Phenomenology and treatment of selective mutism. AB - Selective mutism is a multidimensional childhood disorder in which, according to the most recent studies, biologically mediated temperament and anxiety components seem to play a major role. Several psychotherapy methods have been reported in case studies to be useful, but the disorder is commonly seen to be resistant to change, particularly in cases of long duration. Currently, behaviour modification and other cognitive methods, together with cooperation with the family and the school personnel, are recommended in the treatment of selective mutism. Selective serotonin reuptake inhibitors and selective monoamine oxidase inhibitors have also been reported to be helpful when treating children with selective mutism. At the moment, pharmacotherapy cannot be recommended as the treatment of first choice but if other methods of treatment are not helpful, medication can be included in the treatment scheme. Comprehensive evaluation and treatment of possible primary and comorbid problems that require treatment are also essential. PMID- 11888339 TI - Assessing the efficacy of drugs for the acute treatment of migraine: issues in clinical trial design. AB - Clinical trials of therapies for acute migraine attacks have evolved over the years from open-label, small observational studies to highly structured randomised, controlled trials. The International Headache Society Committee on Clinical Trials in Migraine developed a tool to guide in designing scientifically sound trials. The proof of effect is best achieved in a clinical trial with: clearly defined objectives;a well-characterised study population, identified using well-validated diagnostic tools;proper randomisation and blinding;inclusion of a placebo arm, with proper balancing of patients receiving placebo and those receiving active drug;adequate study power; and appropriate statistical methods. Both parallel and crossover studies may be suitable in clinical trials of antimigraine agents, although the latter are a better choice in patient preference and bioequivalence studies. Although various efficacy measures are used to assess treatment effect, the 2-hour pain free rate (total resolution of pain within 2 hours after an initial moderate to severe headache) is preferred because it is clinically relevant and is relatively 'placebo-insensitive'. Various migraine surveys have indicated that a rapid onset of therapeutic effect is a highly desirable attribute of an antimigraine drug. Therefore, accurate measurements of treatment effect before 2 hours are becoming increasingly emphasised. Consistency of effect across multiple attacks adds to the understanding of the therapeutic efficacy of a test drug. Finally, preference and satisfaction studies allow us to assess patients' global impression of a particular treatment, weighing the positive effects on pain and associated symptoms of migraine against potential adverse effects. PMID- 11888340 TI - Anticonvulsant hypersensitivity syndrome in children: incidence, prevention and management. AB - Anticonvulsant hypersensitivity syndrome (AHS) is a rare, but potentially fatal, adverse reaction that occurs in patients, including children, who are treated with anticonvulsants. During metabolism of the anticonvulsant, toxic arene-oxide compounds are produced. AHS is associated with both cutaneous and systemic symptoms and is associated with multiorgan involvement. Liver damage, in particular, seems to be associated with fatal outcomes. The pathophysiology of AHS is still uncertain but it may be linked to a genetically determined inability to detoxify reactive drug metabolites. The prompt recognition of the first clinical signs of AHS, and the rapid withdrawal of the anticonvulsant, often avoids the progression of symptoms. Pharmacological treatment is essentially based on systemic corticosteroids in association with enteral nutrition, intravenous fluid augmentation, pain relief and ocular care. Intravenous immunoglobulins may also have a possible therapeutic role in some cases. Diagnostic tests, such as patch tests or in vitro assays, for AHS could help to identify patients at risk of developing the syndrome and could represent a first step of primary prevention when applied to relatives of patients. PMID- 11888341 TI - Spotlight on amisulpride in schizophrenia. AB - Amisulpride, a substituted benzamide derivative, is a second-generation (atypical) antipsychotic. At low doses, it enhances dopaminergic neurotransmission by preferentially blocking presynaptic dopamine D(2)/D(3) autoreceptors. At higher doses, amisulpride antagonises postsynaptic D(2) and D(3) receptors, preferentially in the limbic system rather than the striatum, thereby reducing dopaminergic transmission. In patients with acute exacerbations of schizophrenia, the recommended dosage of amisulpride is 400 to 800 mg/day, although dosages < or = 1200 mg/day may be administered. In comparative trials, amisulpride administered within this range (400 to 1200 mg/day) was as effective as haloperidol 5 to 40 mg/day, flupenthixol 25 mg/day and risperidone 8 mg/day in patients with acute exacerbations of schizophrenia with predominantly positive symptoms. Amisulpride was more effective than haloperidol but equally effective as risperidone in controlling negative symptoms. Amisulpride 400 to 800 mg/day was more effective than haloperidol, risperidone and flupenthixol in controlling affective symptoms in these patients. In randomised, double-blind trials involving patients with predominantly negative symptoms of schizophrenia, amisulpride 50 to 300 mg/day was more effective than placebo. Amisulpride is effective as maintenance therapy in patients with chronic schizophrenia. Long term treatment with amisulpride was associated with improvements in quality of life and social functioning. Amisulpride is generally well tolerated. In well controlled trials, the neurological tolerability profile (including ratings on extrapyramidal symptom scales) of amisulpride 400 to 1200 mg/day was superior to that of the conventional antipsychotics (haloperidol or flupenthixol), but was similar to that of the atypical antipsychotic risperidone. At low dosages of amisulpride (< or = 300 mg/day), the incidence of adverse events (including extrapyramidal symptoms) reported with amisulpride was similar to that with placebo. CONCLUSION: In comparative trials, amisulpride 400 to 1200 mg/day showed efficacy in reducing overall symptomatology and positive symptoms similar to that of conventional antipsychotics and newer atypical antipsychotics in patients with acute exacerbations of schizophrenia. Moreover, its effective alleviation of negative and affective symptoms, its lower association with extrapyramidal symptoms and loss of cognitive function than conventional antipsychotics and its long-term efficacy justifies consideration of the use of higher dosages of amisulpride in this group of patients. Consequently, the dosage of amisulpride that is recommended in patients with acute exacerbations of schizophrenia is 400 to 800 mg/day, although dosages < or = 1200 mg/day may be administered. Lower dosages of amisulpride (50 to 300 mg/day) should be considered for the management of patients with negative symptoms of schizophrenia. Amisulpride is a first-line treatment option in the management of schizophrenia in the acute phase and for the maintenance of treatment response. PMID- 11888344 TI - Selective inhibitors of butyrylcholinesterase: a valid alternative for therapy of Alzheimer's disease? AB - The brain of mammals contains two major forms of cholinesterases (ChEs): acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The two forms differ genetically, structurally and in their kinetics. Butyrylcholine is not a physiological substrate in mammalian brains which makes the function of BuChE difficult to interpret. In human brains, BuChE is found in neurons and glial cells as well as in neuritic plaques and tangles in patients with Alzheimer's disease (AD). While AChE activity decreases progressively in the brain of patients with AD, BuChE activity shows some increase. In order to study the function of BuChE, we perfused intracortically the rat brain with a selective BuChE inhibitor. We found that extracellular acetylcholine levels increased 15 fold from 5 nmol/L to 75 nmol/L concentrations, with little cholinergic adverse effect in the animal. Based on these data, we postulated that two pools of ChEs may be present in the brain: one mainly neuronal and AChE dependent; and one mainly glial and BuChE dependent. The two pools show different kinetic properties with regard to regulation of acetylcholine concentration in the brain and can be separated with selective inhibitors. The recent development of highly selective BuChE inhibitors will allow us to test these new agents in patients with AD in order to find out whether or not they represent an advantage for the treatment of patients with AD as compared with selective (donepezil) or relatively non selective (rivastigmine, galantamine) ChE inhibitors presently in use. The association between a BuChE-K variant and AD has not been confirmed in several studies. In conclusion, additional experimental and clinical work is necessary in order to elucidate the role of BuChE in normal brain function and in the brains of patients with AD. In the future, it may be possible that selective BuChE inhibitors will have a role in treatment of patients with advanced AD. PMID- 11888347 TI - Cytomegalovirus and the aging population. AB - Human cytomegalovirus (HCMV) has the largest genome of any virus known to infect man. The virus has evolved many strategies to manipulate the host immune systems and so can remain latent and evade important immune responses. The human host mounts a substantial immune response against the virus, with up to 1% of the virus-specific CD8+ T cells being directed against specific epitopes. Acquisition of HCMV occurs progressively from an early age, and in developed countries the overall seroprevalence is approximately 60%. In contrast, specific communities such as gay men, lower socioeconomic groups and people residing in developing countries have seroprevalence rates that can exceed 90%. It is a widely held belief that successful control of viral infections decreases with increasing age because of a reduction in the capacity of the immune system. Studies in aging populations have shown a specific expansion of the CD8+, CD28- and CD57+ subset of cells in patients who are HCMV-seropositive. Prior infection with HCMV has also been associated with a significantly increased number of CD4+ and CD8+ lymphocytes, as well as cells expressing CD56 and HLA-DR. Thus, HCMV infection can cause substantial perturbations in T cell subsets and these effects persist in the aging population. In the context of solid organ transplantation, older age of both recipients and donors may serve to increase the frequency of donor positive recipient-positive (D+R+) transplants, which have only a moderate risk of HCMV disease. In the context of HIV infection, age has been a dominant risk factor for progression to AIDS and death. At present, it does not appear that this can be explained by lack of immune control of HCMV in the aging population, although studies have identified prior HCMV infection as a risk factor for AIDS and death independent of age. We await further investigations to determine whether the immune control of HCMV in the elderly patient is as effective as in the younger adult, and whether this is linked to pathological consequences. PMID- 11888346 TI - Progressive supranuclear palsy: clinical features, pathophysiology and management. AB - Progressive supranuclear palsy (PSP) is a degenerative condition of unknown aetiology that produces an akinetic-rigid form of parkinsonism characterised by early falls and abnormalities of extraocular movements. Mean age of onset is approximately 63 years, and mean survival from symptom onset is 9 years. Men are much more frequently affected than women. The classic clinical finding is supranuclear ophthalmoplegia, which may not present until late in the illness, if at all. The clinical diagnosis of PSP can be difficult to make, as the sites of pathology are heterogeneous. Structural and functional neuroimaging studies, although not specific for PSP, may be of some assistance in making the diagnosis. The definitive diagnosis of PSP requires the presence of both clinical and neuropathological evidence. Multiple anatomical sites are affected in PSP. The most consistently involved are the subthalamic nucleus, globus pallidus interna and externa, pontine nuclei, periaqueductal grey matter and the substantia nigra. The location of the pathology accounts for the clinical features. The histological hallmark of PSP is the presence of globose neurofibrillary tangles in the affected subcortical nuclei. Neurofibrillary tangles are composed of abnormally phosphorylated tau, a microtubule-associated protein that is involved in maintenance of the cytoskeleton. Abnormalities near or in the gene coding for tau are implicated in the pathogenesis of PSP. The multiple neurotransmitter abnormalities, including those affecting dopamine, acetylcholine, gamma aminobutyric acid and norepinephrine (noradrenaline) systems and pathways, as well as both pre- and post-synaptic pathology, make pharmacological therapy of PSP a challenge. Although an individual patient may respond to a drug, in general patients with PSP have a minimal response and a short duration of sustained benefit. PMID- 11888348 TI - Oral tegafur/uracil. AB - Tegafur is a prodrug of the antineoplastic agent fluorouracil, and is administered in a 1:4 molar ratio with the fluorouracil modulator uracil. Oral tegafur/uracil 300 mg/m(2)/day plus calcium folinate 75 or 90 mg/day for 28 days every 35 days was as effective as intravenous (IV) fluorouracil 425 mg/m(2)/day plus folinic acid 20 mg/m(2)/day for 5 days every 28 or 35 days in the treatment of patients with metastatic colorectal cancer in two large, randomised, nonblind, multicentre trials (n = 816 and 380). Median survival time among patients treated with tegafur/ uracil or fluorouracil was approximately 12 months in both trials. Results from both trials also demonstrated no significant between-group differences in overall response rates among patients treated with oral tegafur/uracil (12 and 11%) or IV fluorouracil (15 and 9%). In elderly patients (aged > or = 70 years) with metastatic colorectal cancer, results from small noncomparative studies showed that treatment with oral tegafur/uracil afforded overall response rates of 12.5 to 29% and was well tolerated. During preoperative treatment with oral tegafur/uracil plus calcium folinate as an adjunct to radiotherapy in patients with stage II or III rectal cancer, the maximum tolerated dosage of tegafur/uracil was 350 mg/m(2)/day (administered 5 days per week for 5 weeks). Among the 15 patients who were followed for 5 to 8 months, three had a complete response to treatment. Treatment with tegafur/uracil was also given postoperatively. The most common adverse events associated with oral tegafur/uracil were anaemia, nausea/vomiting, diarrhoea, thrombocytopenia, mucositis, neutropenia, asthenia, anorexia and abdominal pain. Oral tegafur/uracil was associated with a significantly more favourable tolerability profile than IV fluorouracil in the two large randomised trials. In particular, stomatitis and most adverse haematological events were less frequent. PMID- 11888349 TI - Can angiotensin receptor antagonists be used safely in patients with previous ACE inhibitor-induced angioedema? AB - Angioedema is an uncommon but potentially life-threatening adverse event associated with ACE inhibitor therapy which is believed to be due to potentiation of the vascular effects of bradykinin. Angiotensin receptor antagonists were not expected to produce angioedema, as they do not inhibit the catabolism of bradykinin. However, it is now apparent that angioedema is occasionally associated with angiotensin receptor antagonist therapy and may be more likely to occur in patients who have previously experienced angioedema while receiving ACE inhibitors. Angiotensin receptor antagonists cannot be considered to be a safe alternative therapy in patients who have previously experienced ACE inhibitor associated angioedema. PMID- 11888345 TI - Strategies for management of prostate cancer-related bone pain. AB - Prostate cancer is one of the most common malignancies and a leading cause of cancer-related death in men worldwide. In the majority of cases, prostate cancer metastases to the skeleton, in which case cancer-related bone pain becomes a major cause of morbidity. Androgen ablation is the treatment of choice for securing regression of skeletal metastases in the majority of cases. Intermittent androgen ablation is an attractive alternative, aimed at minimising adverse effects of hormone deprivation but also potentially delaying hormone refractoriness. The development of hormone-refractoriness is heralded by a significant increase in morbidity largely because of escalating bone pain caused by the progression of the metastatic process. Skillful use of analgesics is initially successful but eventually fails to control symptoms. Localised metastases are best treated with local radiotherapy that is rapidly effective. Over the last few years, it has become clear that therapeutic modalities using bone-seeking radionuclides or bisphosphonates have been effective in the palliation of prostate cancer-related bone pain, although not affecting survival. The main limiting factor with the use of radionuclides is bone marrow suppression, also a feature of the very late stages of prostate cancer. Bisphosphonates do not carry this disadvantage. Results of large double-blind, placebo-controlled studies should be awaited, however, before advocating the widespread use of these agents in the management of patients with prostate cancer and skeletal metastases. PMID- 11888350 TI - Improving safety reporting from randomised trials. AB - Randomised clinical trials offer a unique opportunity for capturing safety information under a controlled setting that minimises biases in the comparison of different therapeutic options. Nevertheless, empirical evidence across diverse medical fields suggests that the reporting of safety information in clinical trials is largely neglected and receives less attention compared with efficacy outcomes. An analysis of 192 randomised trials has shown that reasons for withdrawals due to toxicity were specified per study arm in only 46% of the trial reports. Adequate reporting of clinical adverse effects and laboratory-determined toxicity occurred in only 39 and 29% of the trials, respectively, even with lenient definitions of what constitutes adequate reporting. The use of standardised scales for adverse effects is a prerequisite for improved reporting on safety in randomised trials. Safety data need to be collected and analysed in a systematic fashion and active surveillance for toxicity during the conduct of a randomised trial is preferable to passive surveillance. Standardised reporting of safety data does not necessarily require extensive space to accomplish. It is essential to provide numerical data per study arm on each type of adverse effect along with a categorisation of the severity of the adverse effects with an emphasis on severe and life-threatening reactions. The severity grading must be referred to well-known standardised scales and new scales need to be carefully defined. Information on withdrawals due to toxicity is also important to report, along with the specific reasons leading to discontinuation. Tabulation of information may be helpful and rare or not previously reported adverse effects should be described in detail. The availability of newer options such as electronic publication, publication of raw databases, large database research, meta-analytic approaches, and prospective registration of clinical trials and of their databases may further improve the safety insights we can gain from randomised clinical trials. PMID- 11888351 TI - Hypotonic-hyporesponsive episodes following pertussis vaccination: a cause for concern? AB - Vaccine safety has become a major community concern and of particular importance for parents, vaccine recipients and vaccine providers. A hypotonic-hyporesponsive episode (HHE) is a sudden and unexpected episode of loss of tone, unresponsiveness and colour change which uncommonly affects infants and children after vaccination. Although any vaccine may be associated with this adverse event, HHE usually follows administration of a pertussis containing vaccine. There has been renewed interest in this adverse event in the light of community concerns regarding vaccine safety. The focus of this interest has been to formulate an acceptable case definition, to document possible risk factors and to better define the outcome of HHE. In addition, studies have documented the outcome of revaccination of children who have had an HHE. Although much remains to be learnt about HHE it would appear that there are no long-term sequelae and that children who have had an HHE can be revaccinated. Parents should be provided with the available information such that they can make an assessment of the risks and benefits of pertussis vaccination. The benefits of pertussis vaccination still outweigh the risk and universal childhood pertussis vaccination should continue to be advocated. PMID- 11888354 TI - Cost-benefit analysis of the detection of prescribing errors by hospital pharmacy staff. AB - OBJECTIVE: Prescribing errors are a major cause of iatrogenic patient morbidity and therefore interventions aimed at preventing the adverse outcomes of these errors are likely to result in cost reduction. However, it is unclear whether the costs associated with these preventive measures are outweighed by the cost reductions (benefits). Therefore, a study was set up to analyse costs and benefits of detecting prescribing errors by hospital pharmacy staff. DESIGN: During 5 consecutive days in two Dutch hospitals in February 2000 all medication orders, in which prescribing errors were detected, were analysed. A cost-benefit analysis was performed, based on direct medical costs only. The benefit-to-cost ratio was calculated by taking into account the net time hospital pharmacy staff needed for the prevention of the error (this included potential time saving for nursing staff, when an error was prevented by hospital pharmacy staff instead of nursing staff), as well as taking into account the possible consequences of the prescribing error (were the error not prevented). RESULTS: A total of 3540 orders, of which 351 contained prescribing errors (9.9%), were analysed. During the 1-week period investigated, time-investment of the pharmacy staff had net costs of EUR285 (2000 values). During the same period estimated benefits related to this investment were EUR9867. The finding of higher benefits than costs was robust in sensitivity analysis. CONCLUSIONS: From this study it can be concluded that prevention of prescribing errors by hospital pharmacy staff results in higher benefits than the costs related to the net time investment. PMID- 11888352 TI - Effects of psychotropic drugs on seizure threshold. AB - Psychotropic drugs, especially antidepressants and antipsychotics, may give rise to some concern in clinical practice because of their known ability to reduce seizure threshold and to provoke epileptic seizures. Although the phenomenon has been described with almost all the available compounds, neither its real magnitude nor the seizurogenic potential of individual drugs have been clearly established so far. In large investigations, seizure incidence rates have been reported to range from approximately 0.1 to approximately 1.5% in patients treated with therapeutic doses of most commonly used antidepressants and antipsychotics (incidence of the first unprovoked seizure in the general population is 0.07 to 0.09%). In patients who have taken an overdose, the seizure risk rises markedly, achieving values of approximately 4 to approximately 30%. This large variability, probably due to methodological differences among studies, makes data confusing and difficult to interpret. Agreement, however, converges on the following: seizures triggered by psychotropic drugs are a dose-dependent adverse effect; maprotiline and clomipramine among antidepressants and chlorpromazine and clozapine among antipsychotics that have a relatively high seizurogenic potential; phenelzine, tranylcypromine, fluoxetine, paroxetine, sertraline, venlafaxine and trazodone among antidepressants and fluphenazine, haloperidol, pimozide and risperidone among antipsychotics that exhibit a relatively low risk. Apart from drug-related factors, seizure precipitation during psychotropic drug medication is greatly influenced by the individual's inherited seizure threshold and, particularly, by the presence of seizurogenic conditions (such as history of epilepsy, brain damage, etc.). Pending identification of compounds with less or no effect on seizure threshold and formulation of definite therapeutic guidelines especially for patients at risk for seizures, the problem may be minimised through careful evaluation of the possible presence of seizurogenic conditions and simplification of the therapeutic scheme (low starting doses/slow dose escalation, maintenance of the minimal effective dose, avoidance of complex drug combinations, etc.). Although there is sufficient evidence that psychotropic drugs may lower seizure threshold, published literature data have also suggested that an appropriate psychotropic therapy may not only improve the mental state in patients with epilepsy, but also exert antiepileptic effects through a specific action. Further scientific research is warranted to clarify all aspects characterising the complex link between seizure threshold and psychotropic drugs. PMID- 11888356 TI - Antiasthmatic drug delivery in children. AB - Asthma therapy can be administered to children via a number of routes, including oral, inhaled (via a multiplicity of devices), rectal, intravenous, subcutaneous, and intramuscular. The inhaled route is used most often. This can reduce, but never eliminate, systemic absorption. Swallowed aerosolized medication is subject to hepatic first-pass metabolism, but this metabolic route is bypassed by the drug impacting on the airway, including the pharynx. Although there are a large number of studies from a laboratory setting about drug deposition characteristics, there is very little evidence from community-based studies about what families think actually works well in the everyday treatment of the child. However, it is clear that altering the inhaler device can result in marked changes in the dose administered, and any such change should be part of a review of the dose of prescribed medication. Nebulizers are being used much less frequently, and in particular, all but the most severe exacerbations can be treated at least as effectively with equivalent dosages of beta(2)-adrenoceptor agonists from a large volume spacer. PMID- 11888355 TI - Nosocomial pneumonia in pediatric patients: practical problems and rational solutions. AB - Nosocomial pneumonia is a common hospital-acquired infection in children, and is often fatal. Risk factors for nosocomial pneumonia include admission to an intensive care unit, intubation, burns, surgery, and underlying chronic illness. Viruses, predominantly respiratory syncytial virus (RSV), are the most common cause of pediatric nosocomial respiratory tract infections. Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa) are the predominant bacterial pathogens, and are associated with a high mortality rate. Staphylococcus aureus and Staphylococcus epidermidis are the most common Gram-positive bacteria causing nosocomial pneumonia; infections with these organisms have a better outcome than those with Gram-negative organisms. An increasing problem is the emergence of multiresistant Gram-positive and Gram negative nosocomial pathogens. Distinguishing nosocomial pneumonia from other pulmonary processes may be difficult; diagnosis is based on clinical signs, radiological findings, and microbiological results. Recommended empiric therapy should consider factors such as the time of onset of illness, severity of disease, and specific risk factors for nosocomial pneumonia, including use of mechanical ventilation, underlying disease, or recent use of antibacterials. The resident local hospital flora should be considered when selecting therapy for nosocomial pneumonia. Early initiation of appropriate empiric therapy reduces morbidity and mortality. For empiric treatment of bacterial nosocomial pneumonia, an intravenous antibacterial regimen that includes coverage of Gram-negative bacilli and Gram-positive organisms should be used. A carbapenem or ureidopenicillin derivative (piperacillin) plus a beta-lactamase inhibitor should be used where extended spectrum beta-lactamase-producing Enterobacteriaceae are endemic. Therapy should be modified when a specific pathogen and its antimicrobial susceptibility are identified. Effective prevention of nosocomial pneumonia requires infection control measures that affect the environment, personnel, and patients. Of these, hand hygiene, appropriate infection control policies, and judicious use of antibacterials are essential. PMID- 11888353 TI - Clinically significant interactions with drugs used in the treatment of tuberculosis. AB - Clinically significant interactions occurring during antituberculous chemotherapy principally involve rifampicin (rifampin), isoniazid and the fluoroquinolones. Such interactions between the antituberculous drugs and coadministered agents are definitely much more important than among antituberculous drugs themselves. These can be associated with consequences even amounting to therapeutic failure or toxicity. Most of the interactions are pharmacokinetic rather than pharmacodynamic in nature. The cytochrome P450 isoform enzymes are responsible for many interactions (especially those involving rifampicin and isoniazid) during drug biotransformation (metabolism) in the liver and/or intestine. Generally, rifampicin is an enzyme inducer and isoniazid acts as an inhibitor. The agents interacting significantly with rifampicin include anticoagulants, anticonvulsants, anti-infectives, cardiovascular therapeutics, contraceptives, glucocorticoids, immunosuppressants, psychotropics, sulphonylureas and theophyllines. Isoniazid interacts principally with anticonvulsants, theophylline, benzodiapines, paracetamol (acetaminophen) and some food. Fluoroquinolones can have absorption disturbance due to a variety of agents, especially the metal cations. Other important interactions of fluoroquinolones result from their enzyme inhibiting potential or pharmacodynamic mechanisms. Geriatric and immunocompromised patients are particularly at risk of drug interactions during treatment of their tuberculosis. Among the latter, patients who are HIV infected constitute the most important group. This is largely because of the advent of new antiretroviral agents such as the HIV protease inhibitors and the non-nucleoside reverse transcriptase inhibitors in the armamenterium of therapy. Compounding the complexity of drug interactions, underlying medical diseases per se may also contribute to or aggravate the scenario. It is imperative for clinicians to be on the alert when treating tuberculosis in patients with difficult co-morbidity requiring polypharmacy. With advancement of knowledge and expertise, it is hoped that therapeutic drug monitoring as a new paradigm of care can enable better management of these drug interactions. PMID- 11888358 TI - Human recombinant erythropoietin in the prevention and treatment of anemia of prematurity. AB - Human recombinant erythropoietin has been studied extensively as treatment for a variety of anemias. Since in vitro studies showed the primary etiology of the anemia of prematurity to be insufficient serum erythropoietin concentrations, clinical trials have evaluated the administration of human recombinant erythropoietin to preterm infants to treat this indication. These studies were followed by pharmacokinetic determinations in animal models and preterm infants, which revealed that preterm infants required greater doses of human recombinant erythropoietin because of a more rapid clearance and greater volume of distribution. Recent studies have focused on the administration of human recombinant erythropoietin in the first weeks of life to alleviate the anemia caused by excessive phlebotomy losses, and to prevent the anemia of prematurity. In addition, human recombinant erythropoietin has been tried clinically in a variety of neonatal populations in an attempt to decrease or eliminate transfusions. Although much information has been accumulated about the clinical use of human recombinant erythropoietin in preterm infants over the last 15 years, many questions remain unanswered. The evolution of clinical practice in the care of extremely low birthweight infants continues to affect the number of transfusions. It is likely that human recombinant erythropoietin administration in combination with instituting rigorous transfusion guidelines and decreasing phlebotomy losses will have the greatest impact in decreasing transfusion requirements in all preterm and term neonates, regardless of the etiology of their anemia. PMID- 11888357 TI - Corneal ulceration in pediatric patients: a brief overview of progress in topical treatment. AB - Pediatric microbial keratitis is a rare but potentially devastating condition. The condition is similar to adult microbial keratitis, but is often characterized by a more severe inflammatory response. The micro-organisms that cause microbial keratitis in children are similar to the causative agents in adults, with herpes simplex and bacteria being the predominant causative agents, and fungi being less frequent. Of the bacterial pathogens, Pseudomonas aeruginosa, Staphylococcus aureus and alpha-hemolytic streptococci are common. The risk factors for pediatric keratitis include colonization of the eyes during birth and trauma to the cornea. Certain microbial factors involved in microbial keratitis are common to all micro-organisms, including adhesion to the cornea, penetration into the cornea, destruction of the corneal stroma (usually by microbial and/or host proteases), and recruitment of white blood cells to help defend the eye. Specific inflammatory responses that occur during pediatric microbial keratitis are not known in detail, but it is likely that cytokines and polymorphonuclear leucocytes are major factors, as they are in adult microbial keratitis. Treatment for pediatric microbial keratitis is usually the same as treatment for adult microbial keratitis; topical application of antimicrobial agents initially, followed by application of anti-inflammatory agents. With pediatric microbial keratitis, extra care must be taken to ensure nontoxicity due to blood adsorption. New microbial keratitis treatments are being developed and these mainly focus on new antimicrobials, antivirulence agents (such as vaccination against microbial toxins) or specific anti-inflammatory agents. There remains a clear need for increased research into the specific responses during microbial keratitis in children which will help progress new therapies as well as the development of new antimicrobials, especially new antifungal therapies. PMID- 11888360 TI - Design, analysis and presentation of multinational economic studies: the need for guidance. AB - Over the last decade, there has been a proliferation in the number of economic evaluations of pharmaceuticals to meet the growing demand for information about the economic benefits of healthcare technologies. The majority of these studies have been commissioned by pharmaceutical companies for the purposes of drawing attention to the resource and quality-of-life aspects of new or existing products. Such information has become important in overcoming a new obstacle, namely the demonstration of cost effectiveness (the so-called 'fourth hurdle'), in addition to the three well-established criteria of quality, tolerability and efficacy. To ensure the maintenance of standards, guidance for economic evaluations has emerged lately in the form of guidelines, regulations, principles, policies and positions. Drummond outlined three purposes of these guidelines, as follows: as a requirement prior to reimbursement, as statements of methodological standards, and as a statement of ethical standards. Such guidelines are designed to assist both the economic analyst and the decision maker. In laying out the state of the art regarding the methodology of economic evaluation, guidelines assist the analyst in performing high-quality, scientifically valid studies, and assist the decision-maker in properly interpreting and assessing their quality. In response to these growing requirements for cost-effectiveness data globally, it has become increasingly common for economic evaluations to be conducted on an international scale. However, the recommendations in pharmacoeconomics guidelines regarding the manner in which these multinational economic evaluations should be designed, analysed and presented are too limited to be of any real value. This article examines the various issues that must be taken into consideration when conducting multinational studies, and provides a review of the techniques and approaches that have been suggested to date. It concludes with recommendations for potential inclusion in future sets of pharmacoeconomic guidelines. PMID- 11888361 TI - Cost-effectiveness analysis of stratified versus stepped care strategies for acute treatment of migraine: The Disability in Strategies for Care (DISC) Study. AB - BACKGROUND: The Disability in Strategies for Care (DISC) study was the first large randomised controlled trial to compare alternative treatment strategies in the acute treatment of migraine. With 835 patients in its intention-to-treat efficacy analysis, DISC compared a stratified care strategy, where initial therapy was based on clinical need as determined by the Migraine Disability Assessment Scale (MIDAS) and two stepped care strategies (across attacks and within attacks), where first-line therapy with a simple combination analgesic was escalated, if response had been inadequate, to zolmitriptan, a migraine-specific therapy. OBJECTIVE: To report on the cost effectiveness of these three strategies from a societal perspective. STUDY DESIGN AND METHODS: A cost-effectiveness analysis was undertaken using data from the DISC study, and including both health service and productivity costs. Data were collected prospectively on drug usage (main therapy and rescue medication); resource use associated with adverse events was estimated by a clinician blinded to treatment strategy. Health service resource use was costed using UK unit costs (1999 to 2000 values). Data were collected using diary cards on the amount of time patients lost from work, and on reduced effectiveness at work, due to a migraine attack. This facilitated an estimate of the productivity costs associated with the treatment strategies. To assess cost effectiveness, the differences in costs between the strategies were related to the two primary outcome measures in the trial: headache response 2 hours after initial therapy and disability-adjusted time during the first 4 hours after initial therapy. RESULTS: Although the mean health service cost was higher in the stratified care group (mean over 6 attacks of pound 28.25 versus pound 11.74 and pound 23.15 in the stepped care across attacks group and within attacks group, respectively), mean productivity costs over 6 attacks were lower in the stratified group (pound 112.22 versus pound 144.70 and pound 127.53). The total mean cost over six attacks was, therefore, lowest in the stratified care group (pound 138.95 compared with pound 157.19 in the stepped care across attacks group and pound 148.53 in the stepped care within attacks group), although these differences did not reach statistical significance. In terms of headache response, stratified care was statistically significantly more effective than both forms of stepped care. Using disability-adjusted time, stratified care was statistically significantly more effective than stepped care across attacks, but not against stepped care within attacks. CONCLUSION: Given its lower mean costs and higher mean effectiveness, a stratified care strategy, which included zolmitriptan, was the dominant strategy and was unequivocally more cost effective from a societal perspective than either stepped care strategy. When the uncertainty around these means was considered, stratified care had the highest probability of being cost effective. PMID- 11888359 TI - Montelukast: a review of its therapeutic potential in asthma in children 2 to 14 years of age. AB - Montelukast is a cysteinyl leukotriene receptor antagonist which is used as a preventive treatment for persistent asthma in patients > or =2 years of age. In children aged 6 to 14 years montelukast (5 mg/day) treatment resulted in a significant increase in FEV(1) (forced expiratory volume in 1 second, primary clinical outcome) during an 8-week randomized, double-blind trial. Moreover, significant improvements were observed for a range of secondary endpoints assessing symptoms, exacerbation rates, beta-agonist usage and quality of life. Concomitant administration of montelukast (5 mg/day) and inhaled budesonide (200 microg twice daily) resulted in a trend towards an increase in FEV(1) (p = 0.06, primary endpoint) and a statistically significant reduction in both as-needed beta(2)-agonist usage and the percentage of days with asthma exacerbations compared with budesonide plus placebo. No significant differences were observed in asthma-related quality of life between the two groups. During clinical trials both improvements in lung function and reductions in as-needed beta(2)-agonist usage were generally observed within 1 day after initiation of therapy in children 2 to 14 years of age with persistent asthma. Data from a randomized, nonblind trial in 6- to 11-year-old children and a 6-month extension to this trial suggest that both compliance to therapy and patient satisfaction are greater for montelukast than for either inhaled sodium cromoglycate or inhaled beclomethasone. In addition, patients and parents preferred oral montelukast over sodium cromoglycate. In 2- to 5-year-old children with persistent asthma, montelukast (4 mg/day) treatment resulted in significant improvements in a range of outcomes, such as as-needed beta(2)-agonist usage, symptom scores and percentage of days with asthma symptoms, as assessed during a randomized, double blind trial primarily designed to assess tolerability. Data from small randomized, double-blind trials suggest that montelukast reduces exercise-induced bronchoconstriction in 6- to 14-year-old children. Montelukast is generally well tolerated. The frequency of adverse events in montelukast-treated children of all ages was comparable to that in patients receiving placebo. CONCLUSION: Oral montelukast has shown efficacy as a preventive treatment for asthma during clinical trials in children aged 2 to 14 years. The drug offers benefits over more standard therapies such as inhaled sodium cromoglycate and nedocromil in terms of compliance and convenience. In addition, the drug offers significant benefits when added to inhaled corticosteroids (according to secondary endpoints). Montelukast offers an effective, well tolerated and convenient treatment option for children with asthma. PMID- 11888362 TI - Cost-effectiveness analysis of exemestane compared with megestrol in advanced breast cancer: a model for Europe and Australia. AB - OBJECTIVE: To investigate the cost effectiveness of exemestane compared to megestrol in post-menopausal women after tamoxifen failure. DESIGN AND SETTING: A modelling study from the third-party payer perspective in Australia, Belgium, France, Germany, Italy, The Netherlands, Spain and the United Kingdom. METHODS: A model was constructed, based on and driven by data on survival from a clinical study of these agents, including costs for exemestane and megestrol and costs for other treatments. Data from an observational study were used to calculate a country-specific daily cost for the other treatments. Life-years gained was used as the measure of effectiveness. Simulations were performed for 1080 days ('within trial analysis') and for a life-time perspective, in which survival after the end of the trial was assumed to be the same as the trend during follow up. Costs were presented in 1999 values. MAIN OUTCOME MEASURES AND RESULTS: When running the model for 1080 days, the cost effectiveness of exemestane compared to megestrol varied between about EUR5000 and EUR13000 per life-year gained. In Germany it was much lower (EUR1353) due to a higher cost of megestrol. The total expected cost effectiveness (model running until no survivors left) ranged from EUR3700 (Germany) to EUR9100 (The Netherlands). The estimated cost per life-year gained is well within limits generally considered cost effective. CONCLUSIONS: Exemestane is cost effective compared with megestrol for postmenopausal women with progressive advanced breast cancer after therapy with tamoxifen. PMID- 11888363 TI - Health-related quality of life in a UK-based population of men with erectile dysfunction. AB - OBJECTIVE: To measure health-related quality of life (HR-QOL) among a sample of men with erectile dysfunction (ED) in the UK. STUDY POPULATION AND METHODS: A structured questionnaire was mailed to a sample of 5000 men in the UK with ED. The questionnaire included the International Index of Erectile Function (IIEF-5) scale to determine ED severity and the EuroQoL (EQ-5D) questionnaire, a generic HR-QOL instrument. Descriptive information relating to personal relationships and sociodemographics as well as details of comorbidities were also requested. RESULTS: 23% of the sample (n = 1141) returned a completed questionnaire. Of the respondents, 82.2% (n = 939) met the criteria for ED based on the IIEF-5 scale. The mean age of the respondents was 60.4 +/- 24.9 years. There was a gradual convergence of respondents' HR-QOL scores to that of the normal male population as their age increased. The HR-QOL of respondents was significantly poorer than that of the normal population for those under 65 years of age, whereas it was significantly better for those between the ages of 65 and 74 years. Comorbid illness had a significant impact on the HR-QOL of respondents over 44 years of age. Furthermore, the HR-QOL of respondents with multiple risk factors for ED was significantly lower than that of respondents without any risk factors (p < 0.001). The respondents' HR-QOL was significantly poorer compared with the normal male population when stratified by marital status. It was also significantly poorer when stratified by whether the respondents were manual or non-manual workers. CONCLUSION: HR-QOL among men with ED is poorer in those with comorbid illnesses and improves with age. PMID- 11888366 TI - Annual report on access to and utilization of health care for children and youth in the United States--2000. AB - OBJECTIVE: This report provides an update on insurance coverage, use of health care services, and health expenditures for children and youth in the United States. In addition, the report provides information on variation in hospitalizations for children from a new 22-state hospital discharge data source. METHODS: The data on insurance coverage, utilization, and expenditures come from the Medical Expenditure Panel Survey. The data on hospitalizations come from the Database for Pediatric Studies, which is part of the Healthcare Cost and Utilization Project. Both data sets have been prepared by the Agency for Healthcare Research and Quality. RESULTS: Few changes in insurance coverage occurred between 1996 and 1998. About two thirds of American children are covered by private insurance and 19% by public sources; the remaining 15% are uninsured. Of the 71.5% of children who have at least 1 doctor's office visit, the average number of visits was 3.9, but this ranged from 2.7 among the uninsured to 4.2 for those with private insurance. Slightly more than half of children had a prescription, and these averaged 5.4 prescriptions. The majority of children (85%) incur medical expenditures, averaging $1019 for children with any expenditure. Private health insurance was by far the largest payer of medical care expenses for children, even more so than among the general population. However, nearly 21% of expenditures for children's health care were paid out of pocket by children's families. The data also show substantial differences in average length of hospitalization across states, ranging from 2.7 to 4.0 days, and rates of hospital admission through the emergency department, which vary across states from 9% to 23%. Injuries are a major reason for hospitalization, accounting for 1 in 6 hospital stays among 10- to 14-year-olds. In the 10-17 age group, 1 in 7 hospital stays are due to mental disorders. Among 15- to 17-year olds, more than one third of all hospital stays are related to childbirth and pregnancy. CONCLUSION: Children's use of health care services varies considerably by what type of health insurance coverage they have. Expenditures for children entail a substantial out-of-pocket component, which may be quite large for children with major health problems and which may represent a significant burden on lower-income families. Substantial variation in hospitalization exits across states. PMID- 11888367 TI - A national study of commercial health insurance and medicaid definitions of medical necessity: what do they mean for children? AB - OBJECTIVE: To analyze medical necessity standards used by state Medicaid agencies and the largest commercial insurers in the United States on the basis of criteria related to scope of health problems covered and requirements for effectiveness and cost. METHODS: Information was obtained from managed care contract documents used by the 45 state Medicaid agencies enrolling children into managed care organizations and from certificates of coverage used by the largest health maintenance and preferred provider organization insurers in each state. RESULTS: Commercial insurers are more likely than Medicaid agencies to articulate medical necessity standards that limit coverage to treatment for illnesses and injuries and to include stringent requirements for cost and evidence of effectiveness. CONCLUSION: To reduce the discretion retained by insurers in determining medical necessity, particularly around the scope of health problems covered, much greater clarity and uniformity in medical necessity language will be required in the future. PMID- 11888364 TI - Tamoxifen plus chemotherapy versus tamoxifen alone as adjuvant therapies for node positive postmenopausal women with early breast cancer: a stochastic economic evaluation. AB - BACKGROUND: There remains uncertainty around the appropriate choice of adjuvant therapies to offer postmenopausal women with node-positive early breast cancer. OBJECTIVE AND STUDY DESIGN: To present the results derived from a discrete event simulation (DES) model that compared tamoxifen plus chemotherapy versus tamoxifen alone in node-positive postmenopausal women diagnosed with early breast cancer. METHODS: The data populating the model were mainly derived from the existing literature, which was analysed to specify probability distributions describing the uncertainty around the true value of each input parameter. The specified probability distributions facilitated the stochastic analysis of the decision model, whereby distributions of the model's outputs [aggregate costs and quality adjusted life years (QALYs)] were estimated. RESULTS: The baseline results show that the addition of chemotherapy to tamoxifen in this patient group is relatively cost effective (under pound 4000 per additional QALY), but the distribution of the incremental cost-effectiveness ratio shows a wide range, including 10% of observations in which tamoxifen dominates tamoxifen plus chemotherapy. CONCLUSIONS: The results demonstrate the intuitive nature of stochastic evaluations of healthcare technologies, which may ease decision makers' interpretation of cost-effectiveness results. PMID- 11888368 TI - Evidence-based health care coverage for children: proceed with caution. AB - Making health care coverage depend on the existence of valid, applicable research data and positive cost-effectiveness analyses, as managed care contracts are beginning to do, is particularly problematic for children. Because of research challenges specific to children, there are relatively few pediatric data and analyses required under such evidence-based coverage standards. It is too soon to expect major increases from federal efforts to stimulate pediatric health care research. But absence of requisite evidence would entitle a managed care organization or other decision maker to deny coverage on the basis of unproven, negative assumptions about an intervention. In general, population-based evidence is an incomplete basis for decisions on coverage for individual patients. Cost effectiveness analyses are not standardized and may be biased. Purchasers of managed care and policy makers should understand the limits of evidence-based coverage standards. Other uses of evidence may contribute more to systemic improvements of health care. PMID- 11888369 TI - A comparison of health care experiences for medicaid and commercially enrolled children in a large, nonprofit health maintenance organization. AB - BACKGROUND: Proponents of Medicaid managed care have argued that this type of care offers the potential to provide mainstream health care for poor children and the elimination of the 2-tier system of care that has long existed for poor and nonpoor children. However, few studies have attempted to assess whether differences in access, utilization, and satisfaction exist between Medicaid and commercially sponsored children who are enrolled in the same managed care plan. OBJECTIVE: To systematically answer the following research question: Within the same large, nonprofit, group-model health maintenance organization (HMO), how do children enrolled in Medicaid compare with children enrolled commercially across the domains of access, utilization, and satisfaction with care? METHODS: We compared access, satisfaction, and utilization of services between Medicaid and commercially sponsored children enrolled in Kaiser Permanente of Northern California during 1998 through use of a telephone survey and administrative data. Kaiser Permanente is a nonprofit, integrated, group HMO that serves 2.8 million members in more than 15 counties in northern California. The sample for this survey included 510 Medicaid-enrolled children and 512 commercially enrolled children. An overall response rate of 82% was achieved. Bivariate and multivariate analyses were used to compare Medicaid and commercially enrolled children. RESULTS: We found few differences between commercial and Medicaid enrollees across the domains of access, utilization, and satisfaction. Where access differences were present (problems in finding a personal care provider, problems getting care overall, and experiencing 1 or more barriers to care), the differences favored Medicaid-enrolled children. That is, Medicaid enrollees were reported to experience significantly fewer access problems and barriers than commercial enrollees, even after adjustment for confounding factors. Only one difference was found between Medicaid and commercial enrollees across the 6 utilization variables examined (volume of emergency department visits), and no differences were found among the 4 satisfaction and 2 global assessments of care received. Taken together, our results suggest that Medicaid-enrolled children experience as good as or better care than their commercially enrolled counterparts. However, there are other possible explanations for our findings. It may be that families of Medicaid-enrolled children hold their care providers to a lower standard than families of commercially enrolled children, given historic inequities in care between poor and nonpoor families. In addition, some degree of selection bias may be present in our sample, although that is true for both the Medicaid and commercial populations. CONCLUSIONS: Our findings suggest that large commercial HMOs are capable of eliminating the access barriers and stigma traditionally associated with the Medicaid program. However, this conclusion must be tempered with the knowledge that other explanations for our findings may also be at play. PMID- 11888371 TI - Methodologic challenges in developing and implementing measures of quality for child health care. AB - OBJECTIVE: To review the major building blocks in measurement of quality for child health care, with recommendations for future research. METHODS: We describe a framework of building blocks for quality measurement and discuss how an investigator's choices for each component are constrained by the special features of child health care. RESULTS: Methodologic challenges for children's health care include developmental change and dependency on others, fragmentary care and inadequate health care data, unusual care settings, potential for long-term consequences, proxy reporting of outcomes and patient experience, small sample sizes, and lack of evidence that links processes and outcomes of care and of methods for risk adjustment. We cite examples of child-specific measures of quality that illustrate solutions to these challenges. CONCLUSIONS: Children are different from adults, and measures of health care quality for children must differ from those for adults. We suggest future research on measures of quality directed toward overcoming the methodologic problems specific to child health care. PMID- 11888372 TI - Evidence syntheses in child health. AB - Evidence syntheses, also known as systematic reviews, differ from traditional reviews in that they are scientific evaluations of existing studies. Systematic reviews have explicit and reproducible methods and, as with any other scientific endeavor, the result of an evidence synthesis or systematic review can be critically appraised. Many sources for high-quality evidence syntheses now exist, with considerable support from government agencies to develop both the methods and the products of such reviews. Evidence syntheses can increase the efficiency and effectiveness of medical practice but face many hurdles, particularly in child health. These center around 4 areas: lack of high-quality primary studies, the difficulty of finding studies that do exist, the variability and usefulness of the outcome measures in child health, and problems with production and dissemination. Increasing attention to the need for high-quality child health research will help to ameliorate some of these issues, whereas solutions to others are under development or remain elusive. PMID- 11888370 TI - Methodologic challenges in health services research in the pediatric population. PMID- 11888373 TI - Methodologic issues in pediatric outcomes research. AB - Clinicians, health services researchers, and third-party payers, among others, are justifiably interested in the outcomes of pediatric medical care and are, therefore, supportive of research in this area. Pediatric populations pose some unique methodologic challenges for health services researchers. To date, however, many of the approaches, models, and techniques used in pediatric outcomes research have been imported uncritically from experience with adult populations. As a result, some of the most interesting and salient aspects of pediatric outcomes research have yet to be fully developed. These include the following: 1) the problems posed by the dynamics of childhood development, 2) an emphasis on health supervision, 3) the need to see children within the context of a family system and to appreciate the interrelatedness of child health domains, 4) the measurement of the effects of interventions that span sectors, and 5) the paucity of available data sources. This article reviews these problematic areas and argues for a broad conceptual definition of pediatric health, a systems approach to assessing outcomes, and increased interdisciplinary collaboration. PMID- 11888374 TI - Methodologic issues in the conduct and interpretation of pediatric effectiveness research. AB - Effectiveness research represents a number of methodologic challenges not shared with randomized, controlled clinical trials. This practice-based research attempts to translate clinical practices to a wide variety of different practice settings and situations and to diverse patient subgroups. However, because study designs most often used in the conduct of effectiveness research limit the ability to establish firm causal links between medical care and outcomes, it is important to address key methodologic features to generate sound, useable findings. Such features include selection of appropriate outcome measures (with a priori hypotheses linking care to the outcomes chosen), specification of appropriate primary sampling unit, specification of unit of analysis, establishment of appropriate comparison groups, and case-mix adjustment. Conduct of this type of research in pediatrics presents a number of unique methodologic concerns that either do not apply in adult medicine or are particularly acute in pediatrics. To alert policy makers and funders to the unique aspects of pediatric health services research and to provide guidance for the conduct and interpretation of pediatric effectiveness studies, we have organized and described the methodologic issues associated with the specific type of pediatric care under study (eg, specific disease-prevention, "bundled" care for chronic disease, care for problems with social etiologies, etc). We conclude with a summary of the methodologic steps that are critical to the conduct of sound effectiveness research in pediatrics. PMID- 11888377 TI - Factors associated with toilet training in the 1990s. AB - CONTEXT: Few studies have systematically evaluated the factors influencing toilet training in children with normal development. OBJECTIVES: To determine those child, parent, and environmental factors associated with toilet training completion, focusing on the influence of the child's temperament and development. DESIGN AND SETTING: Cross-sectional descriptive study of normal children, ages 15 42 months, attending 1 of 4 pediatric clinics in Milwaukee in 1995 and 1996. METHODS: Demographics for child, parents, and household were surveyed. Temperament was assessed using the Toddler Temperament Scale and the Behavioral Style Questionnaire. Child development was measured using the Bayley Scales of Infant Development II. MAIN OUTCOME MEASURE: Toilet training status was by parental report and was categorized as not trained, not currently training, in training, or training complete. RESULTS: The study population included 496 children, comprising 219 that had not started training, 70 that were not currently training, 148 that were in training, and 59 that were completely trained. The ages at which 50% of the children were predicted to be toilet trained were 35 and 39 months for girls and boys, respectively. In the multivariate regression model, statistically significant factors best predicting toilet training completion were older age, non-Caucasian race, female gender, and single parenthood. Temperament, development, maternal employment, or use of day care were not statistically significant factors. CONCLUSION: Innate factors such as older age, non-Caucasian race, and female gender are the best predictors of completing toilet training (rather than a child's temperament and developmental stage). Day care and maternal employment appear to be unimportant variables. Parents should not be discouraged, because children are completing toilet training at older ages. Research is needed to discover why single parents are more successful at toilet training. PMID- 11888376 TI - Use of a statewide system to improve health and safety in child care facilities. AB - BACKGROUND: The Early Childhood Education Linkage System (ECELS) in Pennsylvania (PA) models ideals of the national Healthy Child Care America (HCCA) Campaign. Little is known about how child care providers use these newly developed statewide systems and about how users compare with nonusers of such a system. OBJECTIVES: Our objectives were 1) to assess knowledge and use of ECELS among child care providers in PA, 2) to compare users and nonusers of ECELS with regard to health concerns, advice-seeking preferences, and infant sleep positioning, and 3) to assess satisfaction among users of ECELS. METHODS: Cross-sectional telephone survey of directors of 400 licensed child care centers (CCCs) and providers of 400 registered family child care homes (FCCHs) in PA. RESULTS: The proportion of children with certain special health care needs mirrored the prevalence in the national child population. Of the facilities surveyed, 88% of CCCs and 71% of FCCHs had heard of ECELS. Among these, 85% had used ECELS's services in the previous 12 months. Significantly more nonusers than users consulted doctors, whereas more users consulted health professionals from government agencies and used printed materials. Of those who enrolled infants, 46% of users and 41% of nonusers reported placing infants on their backs only to sleep. Users who placed infants on their backs were more likely than nonusers to have a written policy about the correct practice (55% and 26%, respectively; P =.02). Overall, 46% of users and 28% of nonusers reported having a sleep position policy (P =.02). Users were at least 95% satisfied with ECELS's services. CONCLUSION: This statewide system reached most child care providers surveyed: more outreach is needed to providers in FCCHs. The health concerns, safety practices, and advice-seeking preferences of child care providers described in this article can inform others who are developing similar collaborative services in each state. Further research on the impact of HCCA programs on health and safety practices (such as correct infant sleep positioning) is warranted. PMID- 11888378 TI - Association of early bilateral middle ear effusion with language at age 5 years. AB - OBJECTIVE: This study tested the hypothesis that children with early persistent middle ear effusion (MEE) are at risk for later language deficit. METHOD: We recruited 698 newborns and monitored them for MEE every 2 to 4 weeks at home until age 3 years. Language skills were assessed on 294 subjects at age 5, while controlling for 8 demographic and environmental factors. Language outcomes at age 5 years were studied as a function of duration of bilateral MEE from birth to age 3 years. RESULTS: A significant relation was found between duration of bilateral MEE and speech sound sensitivity (Carrow Elicited Language Inventory) and articulation (Goldman-Fristoe Articulation). Children's ability to discriminate speech sounds in a quiet environment (Carrow Auditory Visual Abilities Test) was less affected by early prolonged MEE in homes that provided more cognitive stimulation. CONCLUSIONS: These exploratory results indicate that prolonged early MEE may predispose children to language deficits at age 5 years. The language deficits are of small magnitude and may or may not be clinically significant. Language stimulation at home may protect against some of the effects of prolonged MEE. PMID- 11888379 TI - Primary care pediatricians' roles and perceived responsibilities in the identification and management of depression in children and adolescents. AB - OBJECTIVE: To describe primary care pediatricians' 1) approach to the identification and management of childhood and adolescent depression and 2) perception of their skills, responsibilities, and barriers in recognizing and managing depression in children and adolescents. DESIGN AND METHODS: National cross-sectional survey of randomly selected primary care pediatricians that assessed the management of recalled last case of child or adolescent depression, attitudes, limitations to care from barriers and skills, and willingness to implement new educational or intervention strategies to improve care. RESULTS: There were 280 completed surveys about child and adolescent depression (63% response rate). Pediatricians overwhelmingly reported it was their responsibility to recognize depression in both children and adolescents (90%) but were unlikely to feel responsible for treating children or adolescents (26%-27%). Those with most of their practice in capitated managed care were less likely to feel responsible for recognizing depression in either children or adolescents. Forty six percent of pediatricians lacked confidence in their skills to recognize depression in children, and few of them (10%-14%) had confidence in their skills in different aspects of treatment with children or adolescents. Diagnostic, assessment, and management details for their last recalled case of depression in a child or adolescent were provided by 248 of these pediatricians. In addition to referring 78%-79% of the cases to mental health care professionals, 77% of pediatricians provided a wide range of brief interventions. Only 19%-20% prescribed medication. Major factors cited that limited their diagnosis or management were time (56%-68%) and training or knowledge of issues (38%-56%). Fewer pediatricians noted limitations due to insurer or financial issues (8%-39%) or patient issues (19%-31%). The 35% of pediatricians who were motivated to change their recognition and management of suspected depression were significantly more interested in implementing in the future a variety of new strategies to improve care. CONCLUSION: Primary care pediatricians felt responsible for recognizing but not for treating child and adolescent depression. Although the lack of confidence and lack of knowledge and/or skills and time issues are major barriers that limit pediatricians in their treatment of childhood and adolescent depression, pediatricians varied in their readiness to change, with some being more willing to implement new strategies to care for depression. Educational and practice interventions need to focus on how to assist all pediatricians in diagnosis and to prepare these motivated pediatricians to manage depression in primary care settings. PMID- 11888380 TI - Continuity and quality of care for children with diabetes who are covered by medicaid. AB - BACKGROUND: Poor and minority children with Type 1 diabetes mellitus are at increased risk of severe adverse outcomes as a result of their disease. However, little is known about the quality of care that these children receive and which factors are associated with better quality of care. OBJECTIVES: Our objectives were as follows: 1) to describe the utilization of services associated with quality of care for children with Type 1 diabetes mellitus who are covered by Medicaid and 2) to test the hypothesis that increased continuity of primary care is associated with better care for these children. DESIGN: Retrospective cohort study. METHODS: Washington State Medicaid claims data for 1997 were used to determine what proportion of children with diabetes had 1) an inpatient or outpatient diagnosis of diabetic ketoacidosis (DKA), 2) a glycosylated hemoglobin (HgA1c) level that had been checked, 3) a retinal examination, and 4) thyroid function studies. Continuity of care was quantified using a pre-established index. RESULTS: Two hundred fifty-two eligible patients were identified. During the observation year, 20% had an outpatient diagnosis of DKA, 6% were admitted with DKA, 43% visited an ophthalmologist, 54% had their HgA1c checked, and 21% had their thyroid function assessed. Children with high continuity of care were less likely to have DKA as an outpatient (0.30 [0.13-0.71]). Children with medium continuity of care and high continuity of care were less likely to be hospitalized for DKA (0.22 [0.05-0.87] and 0.14 [0.03-0.67], respectively). For preventive services utilization, high continuity of care was associated only with an increased likelihood of visiting an ophthalmologist (2.80 [1.08-3.88]). CONCLUSIONS: The quality of care for Medicaid children with diabetes can be substantially improved. Low continuity of primary care is an identifiable risk factor for DKA. PMID- 11888381 TI - Maternal reports of raising children with chronic illnesses:the prevalence of positive thinking. AB - OBJECTIVE: To examine maternal reports of the positive impact and potential benefits of a child's chronic health condition. DESIGN AND METHODS: A total of 190 mothers and their children with chronic illnesses, including sickle cell disease, cystic fibrosis, diabetes mellitus, and asthma, were recruited from 12 specialty or general pediatric clinics in Baltimore, Md. Standardized interviews were conducted with the mothers and included demographics, condition-related variables, and positive aspects of raising a child with a chronic condition. RESULTS: Eighty-eight percent of the mothers felt better about themselves by learning to manage their child's chronic condition; 70% felt that their families were stronger because of their child's condition; and 80% felt that their family had benefited in some way from having a child with a chronic illness. Ninety eight percent of the mothers endorsed at least 1 positive item; 58% endorsed all 3. CONCLUSIONS: Asking mothers about the positive impact on a family of a child's chronic illness captures an important part of the experience of caregiving. Physicians' recognition and encouragement of this positive outlook may help families continue to face the challenges of raising a child with a chronic illness. PMID- 11888382 TI - Environmental health education in the medical school curriculum. AB - OBJECTIVE: To collect baseline data of environmental history-taking skills and clinical toxicology knowledge and examine the effects of a lecture on environment on students' history-taking skills. METHODS: An anonymous survey was distributed to third-year medical students prior to an asthma lecture that strongly emphasized environmental triggers. Fourteen questions assessed students' practices and attitudes toward environmental history taking. Six multiple-choice questions assessed clinical toxicology knowledge. Histories written by students were reviewed to determine the group's actual performance before and after a lecture on environmental health. RESULTS: Although students reported that an environmental history was important, few asked about environmental history topics other than smoking and pets. Occupational histories were included for adult patients, but few students asked about parental occupations for pediatric patients. Students recognized the correct antidotal therapy for iron and acetaminophen toxicity but were less proficient at identifying clinical features of lead and organophosphate poisoning. Student history performance, when students were considered as a group, was similar to reported performance, with the presence of pets being the only significant postlecture change in history-taking behavior (P =.01). CONCLUSIONS: Students have a positive attitude toward the need for an environmental history, but in self-reported practice and in actual practice, they explore few major environmental history issues. Data were insufficient to prove that one lecture changed history-taking practices. PMID- 11888383 TI - Evaluating medicaid managed care programs for children. AB - Nearly every state has implemented some form of managed care for Medicaid recipients, partly in response to rapid increases in Medicaid expenditures. The unique features of children's health and the differences among states in the implementation and scope of their Medicaid managed care programs provide child health services researchers many opportunities to identify program features that result in favorable health outcomes and those that are less successful. Key stakeholders with interest in this information include state governments and managed care organizations charged with developing and implementing efficient delivery systems, as well as providers interested in understanding the best mechanisms for delivering care to children. This paper outlines potential approaches to evaluating Medicaid managed care programs for children, focusing on identification of appropriate data sources and selection of quality measures encompassing the structure, processes, and outcomes of health care. PMID- 11888384 TI - Market forces and organizational evolution at freestanding children's hospitals in the United States. AB - OBJECTIVES: To describe market forces that affect freestanding children's hospitals, to describe the development of formal business relationships among these hospitals and pediatricians and other health care delivery organizations, and to explore the impact of such changes on the roles and missions of these hospitals. METHODS: All freestanding children's hospitals in the United States in 1991 were identified (n = 44). A survey was mailed to the chief executive officer of each hospital. Data were collected for the period of 1991 through 1996. Twenty nine of the 44 hospitals surveyed responded. RESULTS: Twenty-seven (93.1%) of the 29 hospitals reported an increase in competition and a more advanced stage of market evolution. Twenty-five hospitals (86.2%) developed at least one type of business relationship with pediatricians or another health care organization. Twenty-one (72.4%) developed a network of pediatricians. Seventeen (58.6%) developed a relationship with an adult-focused health care organization. There were no significant differences in teaching, research, or charity care activities between those respondents that developed a pediatric network and those that did not or between those respondents that integrated with adult-focused health care organization and those that did not. CONCLUSIONS: Nearly all freestanding children's hospitals developed new business relationships with physicians and other health care organizations. These new relationships were not associated with any significant changes in teaching, research, or charity care. PMID- 11888386 TI - Frequency of neonatal bilirubin testing and hyperbilirubinemia in a large health maintenance organization. PMID- 11888385 TI - Complementary and alternative medicine use among children in the Washington, DC area. AB - OBJECTIVE: To determine the prevalence and reasons for complementary and alternative medicine (CAM) use among children in primary pediatric care practice in the Washington, DC area. DESIGN: Cross-sectional survey of parents at 4 Children's National Medical Center Pediatric Research Network (PRN) practices from July 1998 through November 1998. Survey information included demographics, child health problems, satisfaction with health care, and CAM use over the past year. RESULTS: Parents completed 348 surveys. Forty percent (138) of parents were CAM users themselves, whereas 21% (72) had treated their child with CAM over the past year. Factors positively associated with child CAM use included parents' use of CAM (P <.0001); greater parent age (P =.0005); greater child age (P =.001); and complaints of frequent respiratory illnesses, asthma, headaches, and nosebleeds. Ethnicity and parental education were not associated with child CAM use. Over 50% of pediatric CAM users reported specific vitamin supplementation, whereas 25% used other nutritional supplements or elimination diets, and over 40% used herbal therapies. Thirty-two percent of CAM users had visited a CAM practitioner; 81% of pediatric CAM users would have liked to discuss it with their pediatrician, but only 36% did so. CONCLUSION: Treatment of children with CAM is common and is frequently undertaken by parents without the knowledge or advice of their pediatrician. PMID- 11888388 TI - A pleasure to work with--an analysis of written comments on student evaluations. AB - OBJECTIVE: Studies assessing rating scales on student evaluations are available. However, there are no data related to the written comments on these evaluations. This study was designed to evaluate these comments. METHODS: A content analysis was performed on the narrative section of pediatric clerks' evaluations. Final evaluations were obtained from 10 outpatient clinical sites staffed by full-time faculty over 14 months. A coding dictionary containing 12 categories (7 linked to clinical skills) was used. RESULTS: One thousand seventeen comments on 227 evaluations were coded. The mean number of comments per evaluation was 4. Learner and personal characteristics were the largest categories. Normative comments, such as "good physical exam," as opposed to more specific comments, such as "complete presentation," predominated in all categories. CONCLUSIONS: Evaluation comments were infrequently related to basic clinical skills and were not often specific enough to lead to effective change in a student's performance. Faculty development is needed to make final evaluation comments more useful for students. PMID- 11888389 TI - Successful use of problem-based learning in a third-year pediatric clerkship. AB - OBJECTIVE: To determine the effect of changing from lectures to a problem-based learning (PBL) curriculum on student knowledge accrual and satisfaction with the didactic portion of a clerkship. DESIGN: Study of cohorts before and after PBL introduction and in comparison with unchanged medicine clerkship. SETTING: Third year pediatric clerkship at 5 clinical sites. PARTICIPANTS: The PBL curriculum was introduced in the 1996 academic year. The 2 classes before the intervention served as historic controls (n = 319), whereas the 2 classes after PBL served as the intervention group (n = 320). INTERVENTION: Small groups of students worked through 6 PBL cases representing common pediatric illnesses. MAIN OUTCOME MEASURES: Knowledge was assessed with the pediatric and internal medicine subject examination of the National Board of Medical Examiners (NBME); student satisfaction was assessed by an anonymous end-of-rotation questionnaire. RESULTS: Scores on the pediatric subject examination improved significantly from means of 69.2 and 70.0 in the historic control group to 73.6 and 74.2 in the PBL cohort (P <.01). Scores on the internal medicine subject exam during the same time periods remained unchanged until the second year after the change (69.1 and 70.2, respectively, before the change; 70.1 and 72.4, respectively, after the change). Analysis of variance results indicated a significant increase in pediatric subject examination scores associated with the introduction of PBL (P <.01). Overall student satisfaction with the pre-PBL lecture series was 3.3, whereas PBL sessions received a score of 4.3. CONCLUSION: Use of PBL in a clinical clerkship was associated with higher scores on the NBME subject examination and increased student satisfaction. These results should encourage the use of PBL during the clinical years. PMID- 11888391 TI - Work-family issues and perceptions of stress among pediatric faculty and house staff. AB - OBJECTIVES: To examine work-family balance issues and predictors of stress related to work-family balance among pediatric house staff and faculty. METHODS: Data were obtained through an anonymous mail survey. Univariate analyses assessed associations between work-family issues (work-related factors that affect work family balance, perceived support, work-family--related stress, and proposed solutions) and the following variables: gender, parental status, working status of spouse, and academic rank. Multiple linear regression examined independent predictors of perceived stress. RESULTS: Fifty percent of the 327 respondents cared for dependent children, and 20% expected to care for an elderly person in the next 5 years. Only 5% strongly agreed that their division or department was concerned about supporting members' work-family balance, and 4% strongly agreed that existing programs supported their needs. Eighty-three percent reported feeling stressed as a result of efforts to balance work and family. Independent predictors of stress included perceived need to choose between career and family, increasing age, dependent children, less support from colleagues and supervisors, and female gender. CONCLUSIONS: Work-family balance issues are responsible for substantial perceived stress. Academic departments should consider a commitment to supporting faculty who are struggling with these issues, including creation of work-family policies and programs, development of mentoring systems, and reexamination of existing expectations for work practices. PMID- 11888390 TI - Development of a Bright Futures curriculum for pediatric residents. AB - OBJECTIVE: To develop a standardized case-based curriculum for pediatric residents on child growth, development, behavior, and adolescent medicine that incorporates the Bright Futures health supervision guidelines. DESIGN: This project included a needs assessment, development of a list of important topics, writing and revising of standardized cases, formative evaluation of cases, and efficacy pilot testing of 2 cases. SETTING: A large pediatric teaching hospital continuity clinic. PARTICIPANTS: Pediatric residents, fellows, and faculty. INTERVENTIONS: Preparation of standardized cases, facilitator training, and resident-led teaching conferences. OUTCOME MEASURES: Learner and facilitator evaluation forms and two 10-item diagnostic skills assessments. RESULTS: During the project, faculty-fellow teams wrote 29 case-teaching modules. All participants gave high ratings to cases, and resident facilitators reported increased comfort with the case discussion method. Resident learners' ability to accurately interpret developmental screening tests and growth charts improved following sessions on those topics. CONCLUSIONS: Further evaluation is required, but these standardized cases appear promising for teaching pediatric residents. The curriculum is now freely available to faculty nationwide. PMID- 11888392 TI - The role of the primary care provider in preventing and treating alcohol problems in adolescents. AB - Adolescents use alcohol more frequently and heavily than all other illicit drugs combined. Given the myriad health, developmental, and social problems associated with alcohol use, it is not surprising that the American Medical Association's Guidelines for Adolescent Preventive Services recommends that adolescents be asked annually about their use of alcohol, and those who report any use during the past year should be assessed further. However, routine alcohol screening of adolescents in primary care and emergency medical settings is not universally applied. In March 2000, the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health and the Agency for Health care Research and Quality jointly sponsored a meeting entitled The Expanding Role of Primary Care in the Prevention and Treatment of Alcohol-Use Disorders. The purpose of the meeting was to bring together researchers, policymakers, clinicians, insurance providers, and medical education specialists to determine the best approaches to increase the involvement of primary care physicians and other health care professionals in screening and intervening for alcohol problems in their patients. The National Institute on Alcohol Abuse and Alcoholism and Agency for Health care Research and Quality believe that the evidence of efficacy for primary care involvement is compelling and are working together to promote the translation of these findings into clinical practice. The following article summarizes what is currently known about adolescent alcohol use and how it can be addressed in primary care settings. It provided the background for the meeting's focus on adolescent issues. PMID- 11888393 TI - About children's oral health needs. AB - Disparities in children's oral health are an important and solvable health problem in the United States today. Many parts of the dental and public health communities are actively engaged in efforts aimed at addressing these disparities. Much progress has been made in explicating the issues, developing the scientific and clinical knowledge base needed for primary prevention and treatment, and beginning the development of new training and community-based approaches. But much more is still needed, and there is a great deal that the pediatric community can do to help. PMID- 11888394 TI - Moving the agenda on pediatric oral health. PMID- 11888395 TI - Barriers to enrollment in a state child health insurance program. AB - OBJECTIVE: To identify barriers to enrollment into Colorado's Child Health Insurance Plan (CHP+) for non-Hispanic (NH), Hispanic (H), and uninsured families. DESIGN: Telephone survey of 1) random samples of families who requested an application but did not complete it (N = 273 NH, N = 159 H) and 2) families with uninsured children identified by random-digit-dial statewide surveys (N = 165). RESULTS: Major reasons for not enrolling included 1) got other insurance (NH 16.5%; H 27.2% P <.01), 2) thought household income was too high to qualify (NH 21.0%; H 11.9% P =.01), and 3) paperwork (NH 13.4%; H 14.7%, P = NS). Of those who thought their income was too high (N = 76, 17.6%), 58.5% appeared eligible based on reported income. Of uninsured families, only 41.7% had heard of CHP+. Of those who had never applied, major remediable reasons included not knowing enough about the program (20.9%) and thinking household income was too high (9.3%). CONCLUSIONS: Effective marketing and education to increase awareness of CHP+ and ensure understanding of eligibility are critical to the success of the program. PMID- 11888396 TI - Gender differences in rates of unintentional head injury in the first 3 months of life. AB - OBJECTIVES: To assess for gender differences in rates of unintentional head injury in infants less than 3 months of age, to assess the circumstances of injury in these patients, and to look for gender-related differences in these circumstances. METHODS: Two separate databases were analyzed. 1) The National Pediatric Trauma Registry (NPTR) was queried for all patients < or = 90 days of age who had been diagnosed with unintentional head trauma between 1990 and 1999. The proportion of males was compared to the expected proportion of 51%, derived from US census data. 2) A prospective cohort of 88 infants < or = 90 days of age who had been treated for unintentional head trauma in an urban pediatric emergency department (ED) was studied. Circumstances of injury and gender-related differences in these circumstances were assessed. RESULTS: In the NPTR database, 600 of 1072 (56%) (95% confidence interval [CI] 0.53, 0.59) infants < or = 90 days of age were boys (P =.001). In the ED cohort, 54 of 88 (62%) (95% CI 0.50, 0.72) subjects were boys (P =.06). In virtually all of the cases described, subjects appeared to be passive participants in the injury. The most commonly reported circumstances of injury were the following: "child left alone on furniture and fell" (n = 39) or "parent dropped child" (n = 27). Boys accounted for 20 (74%) of the subjects in the "parent dropped child" group (P =.04). CONCLUSIONS: Boys outnumber girls among infants less than 3 months of age with unintentional head trauma. These young infants appear to be passive participants in their injuries, which indicates that differences in parenting practices may account for the observed gender differences. PMID- 11888398 TI - Medical education scholarship and ambulatory pediatrics: a review and reflection. PMID- 11888399 TI - The relationship of life stressors and maternal depression to pediatric asthma morbidity in a subspecialty practice. AB - OBJECTIVE: To examine the relationships among demographic characteristics, caregiver life stressors, and depressive symptoms of mothers and their children's asthma morbidity. SETTING: Three pediatric asthma subspecialty programs, 2 in the inner city and 1 in the suburbs. DESIGN: Cross-sectional census sample of caregivers of children with asthma: interviews mostly with mothers (N = 123) regarding their children's asthma symptoms and health care utilization. Information collected on demographics and caregivers' own recent life stressors and depressive symptoms. SUBJECTS: Caregivers of children ages 18 months to 12 years with asthma at their subspecialty visit. MEASURES: Structured interviews: a survey instrument prepared for this study and standardized instruments for depression (Center for Epidemiologic Studies--Depression) and life stressors (Crisis in Family Systems). RESULTS: A total of 32% of respondents' children had high asthma morbidity, 28% intermediate, and 40% low. Caregiver life stressors and depression and the children's sex showed the strongest relationships to asthma morbidity in a model that also included race, residence, and Medicaid status. Children were more likely to have high morbidity if they had caregivers with more depressive symptoms and negative life stressors and if they were female. CONCLUSIONS: Respondents experienced many life stressors and symptoms of depression while managing their children's illness. Caregivers' lives may affect their children's asthma morbidity, offering empirical evidence for the potential value of targeted case management for children in subspecialty care. PMID- 11888400 TI - The use of parent report to assess the quality of care in primary care visits among children with asthma. AB - OBJECTIVE: To determine the accuracy of parent report and the accuracy of the medical record in documenting physician performance of elements of pediatric asthma care in the primary care setting. METHODS: A convenience sample of 79 English-speaking parents of 4--12-year old children with asthma presenting to medical center--affiliated inner-city primary care pediatric clinics in the Bronx, Dallas, and Chicago was enrolled, and the office visit was audiotaped. Parents were interviewed 1--16 days after the visit by telephone. OUTCOME MEASURES: Accuracy of parent report was the primary outcome. The "reference standard" was an independent evaluation of the audiotaped record of the primary care visit. The National Asthma Education and Prevention Program was used as a guide to select data elements to assess quality of pediatric asthma care during primary care visits. RESULTS: Sufficient documentation was significantly (P <.001) less likely to be present in the medical record than in the follow-up interview for each element of care. When these elements were combined into a cumulative score, 71% of parent interviews but only 37% of medical records scored > or = 5 (out of a possible 6), with 29% of medical records scoring < 3. Parents were able to accurately report (concordance of parent data with audiotape reference standard) whether or not the visit had included performance of 5 of the 6 elements of care. CONCLUSIONS: Our study suggests that parent telephone interview within 2 weeks after the visit is more accurate than the medical record for documentation of the quality of asthma care in pediatric primary care visits. The medical record was not sufficient to assess the quality of primary care related to asthma, primarily because of missing data. Therefore, our data suggest that assessing quality of care using the medical record will not only bias the findings in the direction of more deficient care but will also make improvement in care more difficult. Further validation of our strategy for using parent report to assess the quality of care in primary care visits will require its application in a variety of other primary care settings. PMID- 11888401 TI - School readiness among urban children with asthma. AB - BACKGROUND: Children with chronic illnesses, including asthma, are at risk for school problems. Developmental problems, however, may begin before school entry, and the developmental status of preschool children with asthma has not been evaluated. OBJECTIVE: To test the hypothesis that urban preschool children with asthma have lower parent-reported developmental scores compared with children without asthma. METHODS: A comprehensive survey of children beginning kindergarten in 1998 in the urban school system in Rochester, NY, collected parent reports of demographic, medical, and developmental data. We compared children with asthma with and without limitation of activity to children without asthma for motor, language, socioemotional, and school readiness skills and the need for extra help with learning. Linear and logistic regression were used to determine associations between asthma and developmental outcomes. RESULTS: Among the 1058 children in this sample, 9% had asthma, including 5% with asthma with limitation of activity. After adjustment for multiple potential confounding variables, the children with asthma with limitation had lower scores on school readiness skills compared with children without asthma (2.0 vs 2.5, P <.001). Further, the parents of children with asthma with limitation were substantially more likely (P <.05) to describe them as needing extra help with learning (74% vs 56%; odds ratio, 3.2; 95% confidence interval, 1.5--7.8). CONCLUSIONS: Urban preschool children with significant asthma had poorer parent-reported school readiness skills and a greater need for extra help with learning compared with children without asthma. This finding suggests that developmental problems for children with asthma may begin before school entry. PMID- 11888402 TI - Impact of financial incentives on documented immunization rates in the inner city: results of a randomized controlled trial. AB - OBJECTIVE: This study determined the effect of 2 financial incentives---bonus and enhanced fee-for-service---on documented immunization rates during a second period of observation. METHODS: Incentives were given to 57 randomly selected inner-city physicians 4 times at 4-month intervals based on the performance of 50 randomly selected children. Coverage from linked records from all sources was determined for a subsample of children within physician offices. RESULTS: Up-to date coverage rates documented in the charts increased significantly for children in the bonus group (49.7% to 55.6%; P <.05) and the enhanced fee-for-service group (50.8% to 58.2%; P <.01) compared with the control group. The number of immunizations given by these physicians did not change significantly, although the number of immunizations given by others and documented by physicians in the bonus group did increase (P <.05). Up-to-date coverage for all groups increased from 20 to 40 percentage points when immunizations from physician charts were combined with other sources. CONCLUSIONS: Both financial incentives produced a significant increase in coverage levels. Increases were primarily due to better documentation not to better immunizing practices. The financial incentives appeared to provide motivation to physicians but were not sufficient to overcome entrenched behavior patterns. However, true immunization coverage was substantially higher than that documented in the charts. PMID- 11888403 TI - How much does a regional immunization registry increase documented immunization rates at primary care sites in rural colorado? AB - OBJECTIVE: To determine increases in immunization up-to-date (UTD) rates at a rural pediatric practice with the sequential addition of records from other sites in a 2-county region. DESIGN/METHODS: UTD rates for children aged 3 months to 35 months (n = 876) were determined for the index practice and then recalculated after sequential addition of records from 1) the other private practice in the region, 2) 7 public primary care sites, and 3) 2 public health clinics in the region. RESULTS: Adding records from all sites increased documented UTD rates in the index practice from 49% to 64% at 3 months (N = 33, P = 0.025), 50% to 68% at 5 months (N = 38, P = 0.008), 28% to 45% at 7 months (N = 113, P <.01), 29% to 54% at 12 months (N = 200, P <.001), 11% to 35% at 19 months (N = 124, P <.001), and 10% to 33% at 24 months (N = 368, P <.001). CONCLUSIONS: Regional registries will be valuable tools for immunization delivery if there is an ongoing commitment to effective collection of current and historical immunization data. PMID- 11888404 TI - Comparison of risk adjusters for medicaid-enrolled children with and without chronic health conditions. AB - OBJECTIVE: Several capitation payment systems have been developed and implemented recently by public and private insurers as well as by individual managed care organizations. Many pediatricians have expressed concern that methods for establishing capitation rates may not adequately account for the higher expected expenditures for children with chronic health conditions. In this study, we evaluate a demographic- and 4 diagnosis-based models, paying particular attention to their performance for children with chronic health conditions. METHODS: We selected children 18 years of age and under who were enrolled in the Maryland Medicaid Program in 1995 and 1996. We defined the population of children with chronic health conditions using ICD-9 codes. Individual and group-level analyses were utilized to measure the ability of the different risk adjustment models to predict expenditures in 1996 based upon information available in 1995. RESULTS: All 4 diagnosis-based models significantly outperformed the demographic model for children overall and for children with chronic health conditions. Differences between diagnosis-based models were small, especially as the size of test populations increased. CONCLUSIONS: Risk adjustment methods that account directly for health status promise to reduce incentives to exclude children with chronic illnesses from managed care plans and to provide a foundation for more appropriate payments to pediatricians who care for these children. PMID- 11888405 TI - Why should pediatricians be concerned about risk adjustment? PMID- 11888406 TI - Trends and patterns of playground injuries in United States children and adolescents. AB - OBJECTIVE: To determine the prevalence, trends, and severity of injuries attributable to playground falls relative to other common unintentional mechanisms that resulted in an emergency department (ED) visit in the United States. DESIGN AND SETTING: Data from the emergency subset of the National Hospital Ambulatory Medical Care Survey collected from 1992 to 1997 for children <20 years. METHODS: Injury rates and 95% confidence intervals (CIs) were estimated and injury severity scores were computed. RESULTS: There were 920551 (95% CI: 540803 to 1300299) ED visits over the 6-year study period by children and adolescents that were attributable to falls from playground equipment. The annual incidence of visits for playground injuries did not significantly decrease over the course of the study (187000 to 98000, P =.053). Injury visits for playground falls were twice as prevalent as pedestrian mechanisms, but they were less prevalent than visits for motor vehicle-- and bicycle-related injuries. A larger proportion of playground falls resulted in "moderate-to-severe" injury than did bicycle or motor vehicle injuries. Children aged 5 to 9 years had the highest number of playground falls (P =.0014). Playground falls were most likely to occur at school compared to home, public, and other locations (P =.0016). CONCLUSIONS: Playground injury emergency visits have not significantly declined and remain a common unintentional mechanism of injury. Injury visits for playground falls were proportionally more severe than injury visits attributable to other common unintentional mechanisms. Interventions targeting schools and 5- to 9-year-old children may have the greatest impact in reducing emergency visits for playground injuries. PMID- 11888408 TI - Six challenges for academic generalists in pediatric education. PMID- 11888409 TI - Household television access: associations with screen time, reading, and homework among youth. AB - OBJECTIVE: We examined how household factors that mediate television access are associated with screen time (television, videos, movies, and computer and video games), reading, and homework. METHODS: We conducted a self-report survey among 1197 sixth and seventh graders in 10 middle schools in 4 Boston-area communities in 1995. To assess independent associations, SUDAAN linear regressions were calculated to control for respondent characteristics and household access and to account for clustered sampling in the school-based design. RESULTS: Total viewing (television, videos, movies, and computer and video games) averaged 3.35 plus minus 2.2 hours per day. In multivariate regressions, independent direct associations with total viewing were observed for the following categories: youth has a television in the bedroom: 0.64 hours per day (P <.001), never/seldom has family dinners: 0.55 hours (P <.01); no parental limits on television time: 0.48 hours (P <.01); and each additional television outside the youth's bedroom, 0.12 hours (P <.05). Similar results held when television/video/movie use was examined separately from computer/video game use. Youth reported an average of 1.6 plus minus 1.1 hours of reading and homework per day. Parental limits on television time were associated with 0.21 hours more reading per day (P <.01), whereas a television in the bedroom was associated with 0.18 hours less (P <.01). CONCLUSIONS: Reducing intrahousehold television access may enhance clinical, school, and community strategies to reduce youth television viewing and other screen time. PMID- 11888410 TI - Potential health risks of recreational fishing in new york city. AB - BACKGROUND: Fish in the rivers around New York City are contaminated with polychlorinated biphenyls (PCBs) that have been increasingly associated with neurodevelopmental abnormalities. The New York State Department of Health has recommended that the consumption of fish from the rivers around New York City not exceed one meal per month and that no pregnant women or children less than 15 years of age eat any fish from these waters. DESIGN/METHODS: We systematically surveyed anglers at Manhattan fishing sites over a 3-month period to estimate the exposure of anglers, children, and women of childbearing age to PCB-laden fish. RESULTS: One hundred sixty anglers completed the interview. Eighty percent of these anglers were unaware of any advisory to limit fish consumption in order to limit PCB exposure. Most anglers ate the fish they caught. Of the anglers who said that they took the fish home (72.5% of the total sample), 17.7% reported that children regularly eat the fish, and 15.4% reported that an individual had been pregnant while regularly eating the fish. CONCLUSION: Despite state advisories, New York City anglers report high rates of fish consumption by themselves, pregnant women, and children. Although determining the consequences of such consumption will require further study, this represents a worrisome environmental exposure. PMID- 11888411 TI - Should we screen for lead poisoning after 36 months of age? Experience in the inner city. AB - INTRODUCTION: Current lead screening guidelines recommend monitoring lead levels in children under 3 years of age. There are, however, a number of children between the ages of 3 and 6 years who have elevated blood lead levels. Whether these lead elevations are new or chronic has not been examined. OBJECTIVE: To determine the proportion of children with lead levels greater than or equal to 10 microg/dL after their third birthday when all prior testing had been normal. METHODS: Retrospective study based on 39000 venous lead tests obtained between 1993 and 1998. From this group, 2046 children were located who had blood lead levels of less than 10 microg/dL before 36 months and who had a follow-up lead level after 36 months. All lead assays were done by the City of New York laboratories, which had an intrasample variability of 13%. RESULTS: Sixty-six (3.2%) of the 2046 children showed an elevation in blood lead for the first time after their third birthday. The abnormal values ranged from 10 to 25 microg/dL. The majority (72%) of the screen-positive children, however, had lead levels of 10 to 12 microg/dL, and 63.3% of screen-positive children with repeat tests had lead levels that reverted to below 10 microg/dL. CONCLUSIONS: The data indicate that some new cases of lead level elevations did occur after 3 years of age in this 'high-risk' community; however, the current study provides evidence that universal screening for lead poisoning beyond 3 years of age is not warranted in this community as it is not likely to pick up clinically important exposure. PMID- 11888412 TI - A pilot survey of social work services in pediatric primary care programs in urban teaching hospitals. AB - OBJECTIVE: To determine the current status of social work presence in pediatric primary care clinics in urban teaching hospitals. DESIGN: Survey instrument mailed to the medical directors of outpatient pediatrics in the major pediatric teaching hospital of approved residency programs in the 100 largest metropolitan areas in the United States. RESULTS: Sixty responses (60%) were received. Eighty percent of practices reported having on-site social work services, with a median of 14,805 annual clinic visits per social work full-time equivalent. Ninety-five percent of respondents considered on-site social work services in pediatric primary care to be important, whereas half of respondents considered social work services "less than adequate" at their site, and most of these felt this inadequacy had led to additional hospital visits or other adverse outcomes. There were no significant associations of reported adequacy of social work services with any characteristics of hospital, practice, or population. CONCLUSION: Pediatric primary care clinicians at teaching hospitals consider on-site social work services to be important, but most report these services are less than adequate in their practices, and for many, adequacy has declined. PMID- 11888413 TI - Suicide risk following child sexual abuse. AB - OBJECTIVE: To determine the suicide rate and prevalence of suicide attempts and suicidal ideation in 183 young people who had experienced child sexual abuse and to examine variables related to the abuse, which may correlate with suicide attempts or suicidal ideation. METHODS: Adolescents and young adults who had experienced child sexual abuse and individuals from a nonabused comparison group were asked about suicide attempts and suicidal ideation 5 and 9 years after intake to the study. Nine years after the abuse, a national death search was carried out to ascertain the number and causes of death in the 2 groups. Logistic regression was used to assess information on demographic and family functioning variables, the sexual abuse, notifications for other child abuse, criminal convictions, and out-of-home placements that were related to the outcome variables. RESULTS: Young people who had experienced child sexual abuse had a suicide rate that was 10.7 to 13.0 times the national Australian rates. There were no suicides in the control group. Thirty-two percent of the abused children had attempted suicide, and 43% had thought about suicide since they were sexually abused. CONCLUSIONS: Little information seems to be available to clinicians at the time of investigations for child sexual abuse in children that may identify those who are at increased risk of suicide. Abuse by an acquaintance, parental denial, or being angry with the child and not the abuser may predispose to suicide attempts but not necessarily to a completed suicide. PMID- 11888414 TI - Insurance and quality of care for children with acute asthma. AB - BACKGROUND: Increasing attention has been paid to the role of insurance in determining quality and outcomes of care. Pressures to reduce health costs and to improve quality have prompted attempts by managed care organizations to decrease the use of the emergency department (ED) for acute asthma, but performance comparisons between insurance types remain rare. METHODS: We used prospective data from the Multicenter Airway Research Collaboration on 965 children with acute asthma presenting to 36 EDs. We compared measures of quality of pre-ED care, acute severity, and short-term outcomes (length of stay, percent relapse, and percent with ongoing symptoms) across 4 different insurance categories: managed care, indemnity, Medicaid, and uninsured. We used multivariate regression to control for differences in education, estimated income, race/ethnicity, and chronic asthma severity and acute asthma characteristics. RESULTS: Children with managed care and indemnity had similar demographic and asthma characteristics, but these children differed significantly from Medicaid and uninsured patients. Managed care and indemnity insured children had similar ratings on all 7 quality measures, with Medicaid and uninsured children ranking significantly lower on most measures, including (1) percent with primary care provider (PCP) (P <.001), (2) percent using ED as usual site of asthma care (P <.001), (3) percent using ED for prescriptions (P <.001), (4) percent with a ratio of >1 of ED visits to acute office visits within the past year (P =.003), and (5) percent visiting their PCP within the week prior to ED visit (P <.001). Children with managed care were more acutely ill than were indemnity, Medicaid, or uninsured children on presentation to the ED (pulmonary index of 4.6, 4.0, 4.2, and 3.9, respectively, P =.007). There were no significant differences in length of hospital stay, relapse, and ongoing exacerbation. CONCLUSIONS: Our results indicate similar quality of care, greater severity of acute asthma, and no worse outcomes for children with managed care compared to children with indemnity insurance. We found uninsured children to have consistently poorer quality of care than insured patients. PMID- 11888415 TI - Medication use and health care contacts among symptomatic children with asthma. AB - OBJECTIVE: Asthma morbidity and mortality continue to increase despite the availability of improved therapies. Little is known about the degree to which children with asthma use medications and health care services during symptomatic periods. This study documents prospectively the use of medications and health care contacts among children with active asthma symptoms. METHODS: Children age 6 -19 years from 11 primary care settings in upstate New York were eligible for this study if they had 3 or more asthma-related medical visits during the prior year. We collected extensive information on asthma symptoms, medication use, and contacts with health care providers from biweekly phone interviews and daily diaries during a 3-month period. Symptoms were evaluated as the average number of symptomatic days per week. We tabulated the proportion of children using anti inflammatory medications and having health care contacts according to the frequency of their symptoms during this 3-month period. Chi-square and regression analyses were used. RESULTS: One hundred sixty-five children participated (67% White, 24% Black, 9% Other). Sixty-five percent of the children in this sample had an average of more than 2 symptomatic days per week or more than 2 symptomatic nights per month during the 3-month study period and thus had mild persistent to severe asthma. Among these children, 25% received prednisone, and 46% reported the use of an inhaled maintenance medication during the monitoring period. Ten percent of children in this sample experienced an average of 6 or more symptomatic days per week during the study period. Among these highly symptomatic children, only 19% received prednisone, and 56% used a maintenance medication. Further, the proportion of children having contact with a health care provider during this 3-month period was 50% or less, even among the children experiencing the most frequent asthma symptoms. There were no differences in the proportion of children with health care contacts, prednisone use, or maintenance anti-inflammatory use among different gender or race categories or with different insurance types or places of residence. CONCLUSIONS: Even among children experiencing almost daily asthma symptoms, inadequate anti-inflammatory therapy is common, and few contacts with health care providers occur. These children are silently suffering at home and likely are experiencing preventable morbidity. PMID- 11888416 TI - Challenges in long-term health care for children. AB - More children with chronic conditions are surviving than in previous times, and many have serious and significant ongoing health care needs. This paper reviews 1) the population characteristics of children with chronic health conditions in terms of the epidemiology and their sociodemographic profiles; 2) the implications of children's development on their needs, on caretaker roles and responsibilities, on the concept of medical necessity, and on service systems; 3) financing issues and service options for long-term care; and 4) strengths and limitations of existing mechanisms for monitoring the quality of services provided. The following discussion highlights the need for 1) improved data on the numbers of children who need and receive different types of long-term care; 2) better coordination of services and creation of a workable system; 3) a child specific standard of medical necessity in defining service eligibility; 4) better support systems for families caring for children with long-term needs in non institutional settings; and 5) improved consistency in the mechanisms for financing of care for these children. More attention also should be focused on developing ways of monitoring the quality of long-term care provided. Addressing these needs would go a long way toward improving the quality of long-term care for infants, children, and adolescents. PMID- 11888418 TI - Comparison of preventive care in Medicaid managed care and Medicaid fee for service in institutions and private practices. AB - OBJECTIVE: To compare preventive screening for children in Medicaid managed care (MMC) with children in Medicaid fee for service (M-FFS) in private and institutional settings. METHODS: The sample included randomly selected institutions and private practice physicians in New York City. Within setting, children in MMC and M-FFS were sampled randomly and charts reviewed for immunizations and lead and anemia screening. RESULTS: In both institutions and private practices, children enrolled in MMC appeared more likely to be up-to-date than their M-FFS counterparts for immunizations (institution, P <.01; private practice, P <.05), lead screening (institution, P <.01; private practice, P <.01), and anemia screening (institution, P <.01; private practice, P <.01). However, children in MMC had more visits (P <.01) and were followed up for a longer time (P <.01). After controlling for these variables, effects of MMC diminished and only remained significant for screening among private physicians. When considering 10 different attributes of managed care plans, no clear pattern of association with better preventive care services was observed. CONCLUSION: The positive effect of managed care on preventive care services was largely explained by more visits and longer follow-up time; however, there were differences between institutions and private practices, with enrollment in MMC associated with some positive effect on screenings in private practices. PMID- 11888419 TI - Physicians' early challenges related to the pneumococcal conjugate vaccine. AB - OBJECTIVE: To characterize the obstacles faced by physicians regarding administration of a 7-valent pneumococcal conjugate vaccine (Prevnar) to all children younger than 2 years and to high-risk children from 2--5 years of age during the months immediately following national recommendations. DESIGN: Semistructured telephone interviews. PARTICIPANTS: Convenience sample (n = 24) of pediatricians and family physicians. RESULTS: Eighteen physicians were recommending pneumococcal vaccine and 6 were not. Those who were recommending pneumococcal vaccine had encountered resistance from parents and variations in private and public insurance coverage. Physicians who were not recommending pneumococcal vaccine expressed concern about the cost of the vaccine and general caution in adopting new vaccine recommendations. Respondents offered several suggestions for improving the vaccine recommendation process. CONCLUSIONS: This analysis of physicians' early decision making regarding pneumococcal vaccine reflects obstacles to vaccine implementation that may arise with the introduction of other childhood vaccines. PMID- 11888420 TI - Parental employment, family structure, and child's health insurance. AB - OBJECTIVE: To examine the impact of family structure on the relationship between parental employment characteristics and employer-sponsored health insurance coverage among children with employed parents in the United States. METHODS: National Health Interview Survey data for 1993-1995 was used to estimate proportions of children without employer-sponsored health insurance, by family structure, separately according to maternal and paternal employment characteristics. In addition, relative odds of being without employer-sponsored insurance were estimated, controlling for family structure and child's age, race, and poverty status. RESULTS: Children with 2 employed parents were more likely to have employer-sponsored health insurance coverage than children with 1 employed parent, even among children in 2-parent families. However, among children with employed parents, the percentage with employer-sponsored health insurance coverage varied widely, depending on the hours worked, employment sector, occupation, industry, and firm size. CONCLUSIONS: Employer-sponsored health insurance coverage for children is extremely variable, depending on employment characteristics and marital status of the parents. PMID- 11888421 TI - Access to care, unmet health needs, and poverty status among children with and without chronic conditions. AB - OBJECTIVES: To compare insurance coverage, access to care, and unmet health needs of children with and without chronic conditions in a national probability sample of the US population and to examine the role of poverty status in any demonstrated differences between the 2 groups. DESIGN: We analyzed parent-report data on children 0-17 years old from the 1994 National Health Interview Survey Disability Supplement (NHIS-D) and from the health insurance and access to care files of the 1994 Family Resources Supplement to the NHIS. In the NHIS-D, 4452 (14.8%) of the 30032 children were identified as having a chronic condition by a noncategorical method. We compared insurance coverage, access to care, and unmet needs of children with and without conditions overall and also compared them within 3 different income levels relative to the national poverty index: 1) below, 2) within 100%-200%, and 3) >200% above poverty level. RESULTS: In bivariate analyses, children with chronic conditions were more likely to be covered by some type of health insurance (odds ratio [OR], 1.3) and to have a usual provider both for medical ("sick") care (OR, 1.4) and for routine or preventive care (OR, 1.4). They also were more likely to have the same provider for medical care and routine or preventive care (OR, 1.2) and to have seen their health care provider in the last year (OR, 1.8) than children without chronic conditions (all P <.0001). Nonetheless, children with chronic conditions were twice as likely to have had at least 1 unmet need from a list of 4 services that included dental care, prescription medications, eyeglasses, and mental health services (OR, 2.0). They also were more likely to have more than 1 unmet need from the list (OR, 3.1), to have been unable to get needed medical care (OR, 3.1), and to have delayed obtaining medical care because of worry about its cost (OR, 1.8). Children with chronic conditions were at greater risk for unmet needs than were children without conditions across all income levels. The magnitude of the disparity between the groups increased with family income level. Differences persisted even after controlling for sociodemographic variables and insurance status. CONCLUSION: Despite higher levels of insurance coverage and greater access to regular providers of medical and routine care compared with healthy peers, children with chronic conditions are reported by their parents to be less likely than other children to receive the full range of needed health services. The magnitudes of the differences are small, yet the pattern of disadvantage in meeting health care needs among children with conditions compared with healthy peers is consistent across many different variables and it exists across income levels. PMID- 11888422 TI - Recent evidence on the effectiveness of prenatal care. AB - OBJECTIVE: To assess the evidence of effectiveness of prenatal care. DESIGN: A 2 day conference with commissioned papers with summary at the Agency for Healthcare Research and Quality. OUTCOME MEASURES: Effectiveness of prenatal care in preventing prematurity, intrauterine growth restriction, and birth defects and in improving women's health. CONCLUSIONS: Current technology provides limited ability to prevent adverse fetal outcomes but may do much to ameliorate morbidity. However, the latter is not well assessed in data systems. In contrast, the opportunity to improve women's health appears much greater, but doing so is hampered by a number of structural and attitudinal barriers. PMID- 11888423 TI - How much explanation is enough? A study of parent requests for information and physician responses. AB - OBJECTIVE: To examine the relationship between doctor-parent communication patterns and parents' perceptions that they were listened to by the doctor. DESIGN: Cross-sectional, clinic-based survey. Before the visit, parents were asked about the strength of their desires/preferences for being listened to; after the visit, they were asked if their desires were fulfilled and to rate their satisfaction with care. Encounters were audiotaped, transcribed, and coded for parent requests for information and action and physician responses to those requests. Coding was performed using an adapted version of the Taxonomy of Requests by Patients (TORP). Physician responses to parental requests for information were coded as brief, moderate, or prolonged fulfillment or as partially fulfilled, ignored, or denied. SETTING: Two private pediatric practices, 1 community based and 1 university based. PARTICIPANTS: Ten of 13 eligible physicians (participation rate, 77%) and 306 of 356 eligible parents (participation rate, 86%) who sought care for their children's respiratory illnesses. Parents were invited to participate if they spoke and read English and if their child was 2-10 years old, had a chief complaint of cold symptoms, and was seeing one of the participating physicians. Complete data were obtained for 287 doctor-parent encounters (94%). RESULTS: Before the visit, 74% of parents reported that they considered it necessary for the physician to listen to their ideas about their child's illness. Among these parents, 62% (n = 130) reported after the visit that the physician had listened to their ideas. As the proportion of moderate-length responses to parent requests for information increased, parents were significantly more likely to report being listened to (P <.05). Multivariate results indicated a 59% probability of parents reporting that they were listened to when given moderate-length responses, 45% when given brief responses, 39% when given prolonged responses, and 12% when requests for information were only partially fulfilled, ignored, or denied. The length of response to parent requests for information was not related to overall parent satisfaction. CONCLUSIONS: Parents who received moderate-length answers to their questions were most likely to report that they were listened to. Although it is assumed that lengthier, in-depth explanations result in higher satisfaction, this study suggests that more doctor talk does not necessarily constitute better communication. PMID- 11888424 TI - Pediatric residents buckle up: a child safety seat training program for pediatric residents. AB - OBJECTIVES: To assess the effectiveness of supervised installation of child safety seats (CSSs) as a teaching tool for pediatric residents and to evaluate acceptance of this hands-on learning experience. METHODS: Pediatric residents were divided into an intervention group and a control group. All residents completed an initial questionnaire regarding knowledge about CSS use. The intervention group listened to a CSS lecture, viewed a video, and installed CSSs under the supervision of certified CSS technicians. The control group received no intervention. A second questionnaire was administered to all residents. We compared the knowledge gained since the initial questionnaire. The intervention group answered questions regarding their acceptance of this learning experience. RESULTS: Sixty-one residents participated in the study. Most residents had never installed a CSS and felt uncomfortable with their CSS knowledge. The percentage of the intervention group that received a passing score for knowledge increased from 3% initially to 97% on the posttest (P <.001). There was no change in the passing rate of the control group. The intervention group rated the CSS installation session as extremely helpful. CONCLUSION: A hands-on educational program can be an effective, well-accepted method for increasing pediatric residents' knowledge about CSS use. PMID- 11888425 TI - Pediatric hospitalists in Canada and the United States: a survey of pediatric academic department chairs. AB - OBJECTIVES: To document the prevalence and practice patterns of pediatric hospitalists in academic centers in Canada and the United States; to characterize academic pediatric department chairs' definition of the term hospitalist; and to characterize pediatric department chairs' views of the training requirements for pediatric hospitalists. METHODS: A 14-item questionnaire was sent to all 145 pediatric department chairs from Canada and the United States during the fall of 1998. We defined hospitalists as physicians spending at least 25% of their time in inpatient care. RESULTS: Of the 145 eligible pediatric chairs, 128 (89%) responded (United States, 111/126; Canada, 14/16; Puerto Rico, 3/3). Ninety-nine (77%) of 128 pediatric chairs either have (64/128) or are planning to have (35/128) hospitalists in their institutions. Within academic programs with hospitalists, 82% of hospitalists currently work on general pediatric wards. Two thirds of hospitalists teach, 50% provide outpatient care, 50% have administrative duties, and 44% conduct research. One hundred eight (84%) of 128 believe that hospitalists should spend at least 50% of their time in inpatient care. Less than one third (30%) of pediatric chairs believe that hospitalists require training not currently provided in residency. CONCLUSIONS: A large proportion of academic pediatric centers either employed or planned to employ hospitalists in 1998. Pediatric academic department chairs do not see a need for training beyond residency for hospitalists. Further studies should address how pediatric hospitalists affect quality of care, cost, and patient satisfaction. PMID- 11888426 TI - Pediatric hospitalists: what do we know, and where do we go from here? PMID- 11888428 TI - Pediatric jeopardy may increase residents' medical reading. AB - OBJECTIVE: To describe the implementation of a monthly pediatric jeopardy educational intervention (pediatric jeopardy) designed to increase resident reading. METHODS: Pediatric jeopardy, based on the game show JEOPARDY!, was implemented in a pediatric residency training program in September 1997. The questions were derived from the current issues of Pediatrics in Review, Pediatrics, and Pediatrics Review and Education Program. Three residents from each training level competed in teams. Residents' reading habits were briefly surveyed in May 1998 using a 23-item questionnaire. RESULTS: Pediatric jeopardy was implemented and has been continued because both residents and faculty members believed it is a valuable part of the overall residency training program. Some format changes have occurred since the initial implementation of the program. Residents are seen with journals throughout the month, and they speak positively about this conference. The questionnaire response rate was 89.2%. Residents self reported reading an average of 350 minutes per month (5.8 hours) in May 1998. Residents reported that they felt that this program increased their knowledge, motivated them to read, and should be used in other residency training programs. CONCLUSION: Implementation of pediatric jeopardy may increase the amount of overall medical reading reported by the pediatric residents. Pediatric residents reported reading an average of 350 minutes per month. Residents felt this educational intervention was of significant educational value to them personally and should be used in other residency training programs. PMID- 11888429 TI - Commentary on the brief report "Pediatric Jeopardy May Increase Residents' Medical Reading". PMID- 11888430 TI - Faculty and resident attitudes about spirituality and religion in the provision of pediatric health care. AB - OBJECTIVE: To characterize pediatricians' attitudes toward spirituality/religion (S/R) in relationship to the practice of pediatrics. METHODS: Pediatric faculty (n = 100) and residents (n = 65) in an urban academic medical center completed a questionnaire about their attitudes toward and clinical practices related to S/R. Study variables included the strength of personal S/R orientation, attitudes toward S/R, clinicians' discussion of S/R with patients and families, self reported S/R behaviors, the medical conditions that warrant discussion of S/R, and attitudes toward praying with patients if asked to do so. RESULTS: Sixty-five percent of pediatricians felt that faith plays a role in healing, and 76% reported feeling comfortable praying with a patient if asked to do so. Ninety three percent would ask about S/R when discussing a life-threatening illness, and 96% when discussing death and dying. A strong personal S/R orientation was associated with beliefs that the pediatrician should discuss S/R with the patient (P <.01); beliefs that faith plays a role in healing (P <.01); and feelings that patients would like to discuss S/R with their pediatrician (P <.01), that the doctor-patient relationship would be strengthened by discussion of S/R (P <.01), and that physicians should call on an S/R leader for an illness or death (P <.01). Personal S/R orientation was not related to whether physicians reported that they discuss S/R issues with their patients (P =.08). Residents were more likely than faculty to state that it is appropriate to pray with patients if asked to do so (P <.05), and compared with pediatricians who were science majors in college, pediatricians who were nonscience majors in college felt more comfortable praying with patients if asked to do so (P <.01). CONCLUSIONS: In an urban, inner-city, academic medical center, pediatric residents and faculty have an overall positive attitude toward the integration of S/R into the practice of pediatrics. PMID- 11888431 TI - Hospital assessments of children with learning problems: perspectives from special education administrators and hospital evaluators. AB - BACKGROUND: The schooling of 1 of every 8 children is influenced by the Individuals with Disabilities Education Act (IDEA). The IDEA also governs, to a degree, the relationship between interdisciplinary hospital clinics that evaluate many of these children and the schools that provide educational services. Little is known about how this relationship functions or how educational personnel perceive reports from hospital settings. OBJECTIVES: To describe special education administrator and hospital evaluator perspectives on hospital assessments of children with learning problems. METHODS: A survey of attendees at a 1998 meeting of the Massachusetts Association of Special Education Administrators was conducted by use of a questionnaire using closed and open ended questions. Subsequently, focus groups were held with hospital evaluators. RESULTS: Special education administrators describe a high level of frustration with hospital-school relations. They state that hospital reports do not meet the needs of children in the educational setting. They seek closer collaboration with medical evaluators. Evaluators working in hospital settings acknowledge frustration perceived by school personnel. They state that communication with educational personnel improves the quality of their reports and outcomes for the child. CONCLUSIONS: Hospital assessments of children with learning problems are associated with high levels of frustration among special education administrators. These assessments might be made more useful if careful attention were paid to the needs of educators. Greater communication between hospital evaluators and school personnel may increase the usefulness of hospital clinic assessments of children with learning problems. PMID- 11888432 TI - Welfare to work: the effect of a health-care program in child-care centers. AB - OBJECTIVE: Welfare reform has increased pressure on welfare recipients to enter the labor force. When they become employed, former recipients often do not have paid leave that can be used to care for their young children when they are sick. We wished to determine whether an on-site health-care program in child-care centers serving low-income families affected the amount of time parents took off of work to care for mildly ill children. METHODS: We surveyed parents in 6 child care centers with an on-site health-care program and in 2 comparison centers without such a program. To analyze survey results, a regression model including demographic and other variables was used to determine which, if any, variables were associated with time taken by parents from work to care for sick children. RESULTS: Analyzing the variables of employer leave policy, poverty level, age of child, and enrollment in the health-care program, only the variable of health care program enrollment was associated with taking less time from work to care for sick children. CONCLUSION: Health-care programs in child-care settings can help parents meet the health needs of their children while reducing absenteeism from work, thereby contributing to job stability. PMID- 11888433 TI - Identification of children with special health care needs: a cornerstone to achieving healthy people 2010. PMID- 11888434 TI - Framing and using measures of chronic conditions in childhood: a public health perspective. PMID- 11888435 TI - Applying the lessons learned in identifying children with special health care needs: next steps to assure quality care. PMID- 11888436 TI - The national survey of children with special health care needs. AB - CONTEXT: The federal and state-level Children with Special Health Care Needs (CSHCN) programs are vested with the responsibility for planning and developing systems of care for children with special health care needs. To support achievement of this goal, the federal Maternal and Child Health Bureau (MCHB), in partnership with the National Center for Health Statistics (NCHS), has developed a new survey that will provide uniform national and state data on the prevalence and impact of special health care needs among children. PURPOSE: The National Survey of CSHCN is designed to produce reliable state- and national-level estimates of the prevalence of special health care needs using MCHB's definition of CSHCN. It will also provide baseline estimates for federal and state Title V Maternal and Child Health performance measures, for Healthy People 2010 national prevention objectives, and for each state's Title V needs assessment. In addition, it will provide a resource for researchers, advocacy groups, and other interested parties. It is anticipated that this survey will be repeated periodically, thereby making trend analysis possible. METHODS: This survey is being conducted using the State and Local Area Integrated Telephone Survey mechanism, which shares the random-digit-dial sampling frame of the National Immunization Survey (sponsored by the National Immunization Program and NCHS). Using the CSHCN Screener, developed under the auspices of the Foundation for Accountability, 750 children with special health care needs will be identified and selected from each state and from the District of Columbia. Parents or guardians of these children then complete a comprehensive battery of questions on demographics, health and functional status, health insurance coverage, adequacy of health insurance coverage, public program participation, access to care, utilization of health care services, care coordination, satisfaction with care, and the impact of the special need on the family. Data collection began in October 2000 and will continue through March 2002. Summary reports and electronic data files will be available to the public within 6 to 12 months following completion of data collection. CONCLUSIONS: The National Survey of CSHCN will offer a unique data source for individuals and organizations interested in understanding and improving service delivery for CSHCN. It is an accomplishment that reflects the contributions of state and federal Title V programs, family organizations, provider organizations, health services researchers, and the broader maternal and child health community. PMID- 11888437 TI - Identifying children with special health care needs: development and evaluation of a short screening instrument. AB - BACKGROUND: Public agencies, health care plans, providers, and consumer organizations share the need to monitor the health care needs and quality of care for children with special health care needs (CSHCN). Doing so requires a definition of CSHCN and a precise methodology for operationalizing that definition. RESEARCH OBJECTIVES: The purpose of this study was to develop an efficient and flexible consequence-based screening instrument that identifies CSHCN across populations with rates commensurate with other studies of CSHCN. METHODS: The CSHCN Screener was developed using the federal Maternal and Child Health Bureau (MCHB) definition of CSHCN and building on the conceptual and empirical properties of the Questionnaire for Identifying Children with Chronic Conditions (QuICCC) and other consequence-based models for identifying CSHCN. The CSHCN Screener was administered to 3 samples: a national sample of households with children (n = 17985), children enrolled in Medicaid managed care health plans (n = 3894), and children receiving Supplemental Security Income (SSI) benefits in Washington State (n = 1550). The efficiency, impact of further item reduction, and flexibility of administration mode were evaluated. Rates and expected variation in rates across demographic groups of children positively identified by one or more of the 5 CSHCN Screener item sequences in each sample were examined and multinomial logistic regression analysis were conducted to evaluate the effect of child characteristics in predicting positive identification. RESULTS: The CSHCN Screener took approximately 1 minute per child to administer by telephone and 2.1 minutes per household. During self administration, over 98% of respondents completed each of the 5 CSHCN Screener item sequences, and respondents accurately followed each of the item skip patterns 94% of the time. Mailed surveys and telephone-administered surveys led to similar rates of positive identification in the same sample. Two Screener items would have identified 80%-90% of children positively identified as CSHCN across the study samples, although using only 2 items eliminates some children with more complex health needs. Rates of children identified by the CSHCN Screener varied according to age, sex, race/ethnicity, health status, and utilization of health services. CONCLUSIONS: Results of this study indicate that the CSHCN Screener requires minimal time to administer, is acceptable for use as both an interview-based and self-administered survey, and that rates of children positively identified by the CSHCN Screener vary according to child demographic, health, and health care-need characteristics. The CSHCN Screener provides a comprehensive yet parsimonious and flexible method for identifying CSHCN, making it more feasible than existing measures for standardized use across public agencies, health care plans, and other users. PMID- 11888438 TI - Comparison of the children with special health care needs screener to the questionnaire for identifying children with chronic conditions--revised. AB - BACKGROUND: The Children with Special Health Care Needs (CSHCN) Screener is an instrument to identify CSHCN, one that is based on parent-reported consequences experienced by children with ongoing health conditions. Information about how this instrument compares to other methods for identifying CSHCN is important for current and future uses of the CSHCN Screener. RESEARCH OBJECTIVES: The goal of this study was to assess the level of agreement between the CSHCN Screener and the Questionnaire for Identifying Children With Chronic Conditions--Revised (QuICCC-R) and to describe the characteristics of children in whom these methods do not agree. METHODS: The CSHCN Screener and the QuICCC-R were administered to 2 samples: a random sample of parents of children under age 18 years through the first pretest of the National CSHCN Survey (n = 2420) and a random sample of children under age 14 years enrolled in a managed care health plan (n = 497). Information on specific conditions and needs for health services were collected for children identified by one or both instruments in the national sample. Data from the administrative data-based Clinical Risk Groups (CRGs) were collected for all children in the health plan sample. The proportions of children identified with the CSHCN Screener and the QuICCC-R were compared, the level of agreement between these 2 methods was assessed, and the health service needs of children identified by the QuICCC-R but not the CSHCN Screener were evaluated. RESULTS: In both study samples, the CSHCN Screener agreed with the QuICCC-R approximately 9 out of 10 times on whether or not a child was identified as having a special health care need. Compared to the CSHCN Screener, the QuICCC-R identified an additional 7.6% and 8.5% of children as having special health care needs in the national and health plan samples, respectively. Compared to children identified by the QuICCC-R only, the odds were 12 times greater that children identified by both the CSHCN Screener and the QuICCC-R needed health care services, 6 times greater that parents named a specific chronic health condition, and 9 times greater that children were identified with a chronic condition using the CRG algorithm. Study design and purposeful differences in question design or content account for most cases in which children are not identified by the CSHCN Screener but are identified using the QuICCC-R. CONCLUSIONS: The brief CSHCN Screener exhibits a high level of agreement with the longer QuICCC-R instrument. Whereas nearly all children identified by the CSHCN Screener are also identified by the QuICCC-R, the QuICCC-R classifies a higher proportion of children as having special health care needs. PMID- 11888440 TI - Comparing different definitions of chronic conditions in a national data set. AB - OBJECTIVES: To compare the application of 4 different definitions of chronic conditions and disabilities in children using a large national database. METHODS: The study used data on children 0-17 years of age from the 1994 and 1995 National Health Interview Survey (NHIS) Core Interview and Disability Supplement. We selected items to create algorithms that operationalized 4 extant definitions and compared the proportions of children identified by each. For each definition we conducted bivariate and logistic regression analyses of age and sociodemographic and health-related characteristics potentially related to presence of disability. We then compared the results across definitions to determine if they were equally likely to identify children across a full range of sociodemographic variables, ages, and health-related characteristics. We also examined the level of agreement among definitions and determined the degree to which they identified the same or different children. RESULTS: Using a single data set, the 4 operational definitions identified somewhat different proportions of children as having chronic conditions or disability overall (the range was 13.7%-17%). However, the ways in which the definitions identified children along a spectrum of variables did not differ substantially across the definitions. The percent agreement in identification between any two individual definitions was > or = 95%. In general, a higher proportion of children were identified among males, whites, those in older age groups, and among those whose parents had lower levels of education and those who were living below the poverty level. Although the same set of variables identified children with conditions who fit all 4 definitions compared with children who met the criteria for 1 to 3 definitions, higher rather than lower parent education was associated with agreement across all definitions. CONCLUSIONS: Little has been known about the comparability of different definitions for identifying children with chronic conditions, special health care needs, and disability. Our findings suggest substantial overlap in the numbers and characteristics of the children that several different conceptual definitions identify. It remains unclear whether subgroups with higher or lower proportions of children with chronic disorders represent a substantive finding or whether they result from methodological problems shared by the ways in which the definitions are operationalized. PMID- 11888439 TI - Comparing a diagnosis list with a survey method to identify children with chronic conditions in an urban health center. AB - OBJECTIVE: To compare a diagnosis list to the Questionnaire for Identifying Children with Chronic Conditions (QuICCC) to assess their relative usefulness as measures for identifying children with chronic conditions. METHODS: Comparison of health encounter data and survey data for a cohort of 304 children aged 0-18 years at an urban health center affiliated with a teaching hospital. We used 2 strategies to identify children with a chronic condition: 1) identification by the existence of an encounter with an International Classification of Diseases, Ninth Revision code indicating a chronic condition and 2) identification by the QuICCC. We compared the characteristics of children identified by the diagnosis list with those of children identified by the QuICCC. RESULTS: This population had high rates of chronic conditions, with 44% identified by the diagnosis list and 36% identified by the QuICCC. These 2 methods jointly identified 66% of children, yet only half (53%) of the children who had a diagnosis of a chronic condition in the encounter data were identified by the QuICCC. Asthma, anorexia, developmental delay, and adjustment reaction were among the common chronic conditions for children identified by the diagnosis list approach only. CONCLUSIONS: We found only moderate concordance among the children identified as having chronic conditions by a diagnosis list and by the QuICCC in this high-risk urban population. These different results indicate that encounter data and survey approaches do not serve as simple substitutes for identifying children with chronic conditions for clinical or monitoring purposes. PMID- 11888441 TI - Identifying and classifying children with chronic conditions using administrative data with the clinical risk group classification system. AB - OBJECTIVE: To identify and categorize children with chronic health conditions using administrative data. METHODS: The Clinical Risk Groups (CRGs) system is used to classify children, aged 0-18 years, in a mid-sized health plan into mutually exclusive categories and severity groups. Enrollees are categorized into 9 health status groups--healthy, significant acute, and 7 chronic conditions--and are then stratified by severity. Utilization is examined by category and severity level based on eligibility and claims files for calendar year 1999. Only children enrolled for at least 6 months (newborns at least 3 months) are included. RESULTS: This analysis of 34544 children classifies 85.2% as healthy, including 19.6% with no claims; 5.2% with a significant acute illness; 4.6% with a minor chronic condition; and 4.9% with a moderate to catastrophic chronic condition. The average number of unique medical care encounters per child increases by chronic condition category and by severity level. Compared to national prevalence norms for selected conditions, CRGs do well in identifying patients who have conditions that require interaction with the health care system. CONCLUSIONS: CRGs are a useful tool for identifying, classifying, and stratifying children with chronic health conditions. Enrollees can be grouped into categories for patient tracking, case management, and utilization. PMID- 11888442 TI - The generation gap in modern surgery. PMID- 11888443 TI - Why the numbers are dropping in general surgery: the answer no one wants to hear- lifestyle! PMID- 11888444 TI - The generation gap in modern surgery: a new era in general surgery. AB - General surgery remains one of the most respected residencies available to medical students today. At the same time, the number of medical school graduates applying for general surgery residencies continues to decline. Despite this decline, we still search for those individuals exhibiting qualities shared by general surgeons who excel. Although the field of general surgery is constantly evolving, these qualities remain the same. While intellect and good technical skills are essential, they alone do not ensure success as a surgery resident. Confidence, stamina, tenacity, and patience are imperative. Good leadership, motivational, and decision-making skills are also vital characteristics. Finally, the importance of dedication to patient care cannot be overstated. Because disease, injury, and pain are not scheduled events, general surgery residents regularly spend long hours, day and night, caring for and worrying about their patients. To ensure that we continue to add a sufficient number of general surgery residents each year without lowering our standards, those of us in general surgery must attempt to determine why fewer medical school graduates are applying for general surgery residencies. Then we must find ways in which we can improve the residency programs and, probably more important, emphasize to students the things about this field that led us to devote our lives to its practice. PMID- 11888445 TI - Contemporary trends in student selection of medical specialties: the potential impact on general surgery. AB - HYPOTHESIS: Lifestyle is a priority among senior medical students when selecting a career specialty. The trend toward controllable lifestyle vs noncontrollable lifestyle specialties is affecting the number of students desiring a career in general surgery. DESIGN: The Medical Student Graduation Questionnaire is published and distributed by the Association of American Medical Colleges to all US medical schools for senior medical students to complete before graduation. The results from the survey are published each year in the All Schools Report. We evaluated these reports to track the percentage of students pursuing a career in general surgery during the past decade. The National Resident Matching Program also publishes a report each year outlining the match results. We reviewed these results from 1978 through 2001 and used them to determine the percentage of students choosing to enter general surgery. MAIN OUTCOME MEASURES: First choice of specialty among graduating senior students from US medical schools; positions matched by US and foreign medical students and students from osteopathic medical schools; factors that influenced the decision-making process in choice of specialty; and factors that influenced students to change their mind from one career to another. RESULTS: An established trend of decreasing interest in general surgery exists and has the potential to affect the number of positions that are filled each year in the match. Linear projections confirm that, should the current trend continue (negative slope; P =.01), by 2005 only 4.8% of US graduating senior medical students will be interested in general surgery. This established trend of decreasing interest in general surgery, which began in the early 1980s, did not affect the match until 2001, when the number of positions offered exceeded the number of students interested in general surgery. At present, the specialty of general surgery is at risk for significant numbers of positions remaining unfilled. Our match projections estimate that for 2005, only 76.6% of positions will be filled by US senior students (negative slope; P =.001). CONCLUSIONS: If the trend continues, the students matching in general surgery will not be as competitive as in years past, and there will be a potential shortage of these specialists in the United States. PMID- 11888446 TI - The generation gap: perspectives of a program director. PMID- 11888447 TI - "Halstedian 2" residency training: bridging the generation gap. PMID- 11888448 TI - The modern medical school graduate and general surgical training: are they compatible? PMID- 11888449 TI - Effective prevention of adhesions with hyaluronate. AB - HYPOTHESIS: Hyaluronate sodium in the form of a bioresorbant membrane reduces the development of intra-abdominal adhesions frequently found after implantation of synthetic mesh in the context of surgical hernia repair. DESIGN: The effect of hyaluronate on the formation of adhesions was evaluated when applied laparoscopically as a bioresorbant membrane to protect the peritoneal surface of a synthetic mesh. SETTING: Experimental animal model. INTERVENTIONS: A peritoneal defect 5 cm in diameter was bilaterally created in the abdominal wall of each of 9 pigs by laparoscopy. A polypropylene mesh was fixed with clips onto these defects on both sides. In each of the animals, only on one side, the synthetic mesh was also covered by a hyaluronate membrane. MAIN OUTCOME MEASURES: The incidence and severity of adhesions (grade 0-4, where 0 indicates no adhesion; 1, filmy avascular adhesions; 2, vascular adhesions; 3, cordlike fibrous adhesions; and 4, plain fibrous adhesions) were determined after 45 days, comparing treated and untreated sides by autopsy results and histological features. RESULTS: Adhesions, mainly grades 3 and 4, occurred in 7 of the 9 animals in those meshes not covered by hyaluronate; 2 untreated animals did not develop adhesions. On the other hand, only 1 of the 9 animals developed adhesions (grade 2) at the mesh concealed by the hyaluronate membrane. CONCLUSIONS: The bioresorbant hyaluronate membrane significantly reduced the formation of peritoneal adhesions (1-sided sign test, P<.05) induced by the insertion of a polypropylene mesh, when compared with the contralateral implants not protected by hyaluronate. Thus, hyaluronate membranes are efficient for reducing the incidence of peritoneal adhesions. PMID- 11888450 TI - Determinants of mortality in patients with severe blunt head injury. AB - CONTEXT: Head injury is the leading cause of traumatic death in the United States. HYPOTHESIS: A set of clinical parameters available soon after injury can be used to accurately predict outcome in patients with severe blunt head trauma. DESIGN: Validation cohort study. SETTING: Urban level I trauma center. PATIENTS AND METHODS: Data from patients with severe blunt head injury, as defined by inability to follow commands, were prospectively entered into a neurosurgical database and analyzed. The impact on survival of 23 potentially predictive parameters was studied using univariate analysis. Logistic regression models were used to control for confounding factors and to assess interactions between variables, whose significance was determined by univariate analysis. Goodness of fit was calculated with the Hosmer-Lemeshow c statistic. The predictability of the logistic model was evaluated by measuring the area under the receiver operating characteristic curve (AUC). RESULTS: Logistic regression analysis revealed that 5 risk factors were independently associated with death. These variables included systemic hypotension in the emergency department, midline shift on computed tomographic scan, intracranial hypertension, and absence of pupillary light reflex. A low Glasgow Coma Scale score and advanced age were found to be highly correlated risk factors that, when combined, were independently associated with mortality. The model showed acceptable goodness of fit, and the AUC was 80.5%. CONCLUSIONS: Systemic hypotension and intracranial hypertension are the only independent risk factors for mortality that can be readily treated during the initial management of patients with severe head injuries. When used together, Glasgow Coma Scale score and age are significant predictors of mortality. PMID- 11888451 TI - Clinical node-negative thick melanoma. AB - BACKGROUND: Patients with T4 N0 M0 melanoma are considered at high risk for having occult metastases, and adjuvant therapy is usually recommended. HYPOTHESIS: Long-term survival in patients with thick melanoma is not universally poor. DESIGN: A retrospective study. SETTING: University teaching hospital. PATIENTS: We evaluated clinical node-negative thick (> or = l4.0 mm) melanoma in 151 patients who received their primary definitive surgical treatment in our department. None of these patients received any adjuvant therapy. RESULTS: Median follow-up was 44 months; median thickness, 5.5 mm. Median overall (OS) and disease-free survivals (DFS) were 70 (5-year survival, 52%) and 51 months (5-year survival, 47%), respectively. Patients with node-positive disease faired significantly worse than did those with node-negative disease. Median OS and DFS for patients with node-positive disease were 49 and 32 months (5-year survival, 35%), respectively, compared with 209 (5-year survival, 61%) and 165 months (5 year survival, 56%), respectively, for patients with node-negative disease. Similarly, OS and DFS were significantly lower when the primary tumor had at least 5 mitoses/mm(2) or was located in the head and neck region. After multivariate analysis, status of the lymph nodes was the most predictive variable for OS and DFS. CONCLUSIONS: The thickness of melanoma, by itself, should not be used as a criterion for adjuvant therapy. Other prognostic factors should be considered. PMID- 11888452 TI - Treatment of postoperative peritonitis of small-bowel origin with continuous enteral nutrition and succus entericus reinfusion. AB - HYPOTHESIS: Proximal intestinal stomas established by the exteriorization of leaking anastomosis in the presence of peritonitis can be used to reinfuse succus entericus and provide adequate enteral nutrition. DESIGN: Retrospective analysis of prospectively gathered data from a cohort of consecutive patients admitted between January 1993 and December 1999 for postoperative peritonitis requiring laparotomy and the construction of one or more small-bowel stomas. SETTING: Tertiary referral center with a surgical intensive care unit experienced in the treatment of intra-abdominal sepsis and succus entericus reinfusion. PATIENTS: Twenty-one consecutive patients with postoperative peritonitis originating from a jejunal or ileal leak. We excluded patients with established enterocutaneous fistulae, abscesses amenable to percutaneous drainage or other conservative treatments, and postoperative peritonitis caused by ileocolic or ileorectal anastomosis. INTERVENTIONS: Early laparotomy with exteriorization of small-bowel leak(s), and continuous enteral nutrition (CEN) and succus entericus reinfusion (SER) via the distal portion of the stoma until gastrointestinal continuity was restored. MAIN OUTCOME MEASURES: Feasibility of CEN and SER with temporary, diverting small-bowel stomas and their associated postoperative morbidity and mortality rates. RESULTS: One patient died, and 14 experienced complications. For technical reasons, CEN and SER were discontinued early on in 7 patients. The mean duration of CEN and SER was 58 days and 61 days, respectively. Enteral feedings allowed the suppression of central venous access after a median of 28 days, with 82 days as a median time to restoration of intestinal continuity. CONCLUSIONS: Although the exteriorization of small-bowel leaks with CEN and SER is generally feasible and effective in the treatment of critically ill patients with peritonitis secondary to small-bowel leaks, it is associated with significant morbidity and mortality, in part relating to patients' underlying diseases. PMID- 11888453 TI - Video-assisted vs conventional thyroid lobectomy: a randomized trial. AB - HYPOTHESIS: Video-assisted thyroid lobectomy improves the cosmetic outcome of the cutaneous scar and reduces postoperative pain. PARTICIPANTS: Patients admitted to the Division of Endocrine Surgery of the Universita Cattolica del Sacro Cuore, Rome, Italy, between March 1999 and December 2000 who were candidates for thyroid lobectomy because of a single, small (< or = 3 cm) thyroid nodule were considered eligible. Of the 62 patients who were randomized, 31 underwent conventional thyroid lobectomy (COS group), and 31 underwent video-assisted surgery without carbon dioxide neck insufflation (VAS group), a new technique created by the authors. RESULTS: The cosmetic outcome was evaluated by scoring patients' satisfaction with their scars. Satisfaction was higher in the VAS group (mean +/- SD, 9.2 +/- 0.5) than the COS group (mean +/- SD, 5.8 +/- 0.7) (P<.001). Postoperative pain in the first and second days after surgery was lower in the VAS group (mean +/- SD, 1.8 +/- 0.2 and 1.2 +/- 0.1, respectively) than in the COS group (mean +/- SD, 6.2 +/- 0.2 and 5.8 +/- 0.2, respectively) (P<.001). There were no significant differences in complications (eg, bleeding, wound infection, permanent recurrent nerve palsy). Postoperative hospital stay was lower in the VAS group (mean +/- SD, 1.1 +/- 0.1 days) than in the COS group (mean +/- SD, 2.2 +/- 0.2 days) (P<.05). CONCLUSION: Video-assisted thyroid lobectomy is a valid alternative to conventional surgery in patients with single, small nodular thyroid lesions. PMID- 11888455 TI - Spontaneous intramural small-bowel hematoma: clinical presentation and long-term outcome. AB - HYPOTHESIS: To review our experience with the treatment of patients with nontraumatic spontaneous intramural small-bowel hematoma. Our hypothesis was that this condition resolves spontaneously in most patients. DESIGN: A retrospective review of the records of 13 patients with nontraumatic spontaneous intramural small-bowel hematoma who presented to Mayo Clinic (Rochester, Minn; Scottsdale, Ariz; and Jacksonville, Fla) between January 1, 1983, and December 31, 2000. SETTING: A tertiary care medical institution. PATIENTS: Mean age at presentation was 64 years (8 men, 5 women). Patients presented with abdominal pain (13 patients), intestinal obstruction (11 patients), and biliary obstruction (1 patient). Mean duration of symptoms was 4 days. Eight patients were receiving anticoagulant therapy (mean international normalized ratio, 11.6). Only 1 patient was anemic at presentation, but 11 patients became anemic during hospitalization. Computed tomography established the diagnosis in all patients. MAIN OUTCOME MEASURES: Short- and long-term outcomes obtained from clinical records and telephone interviews. RESULTS: Single and multiple hematomas were present in 11 patients and 2 patients, respectively. Two patients had an exploratory operation, but no bowel resection was performed. The other 11 patients were managed with bowel rest. Two patients died of sepsis related to their coexisting medical conditions, and 11 patients left the hospital without short-term complications. At follow-up (mean, 35 months), 4 patients had died of unrelated causes, and 7 were alive; none had recurrence of bowel hematoma or intestinal obstruction. CONCLUSION: Nonoperative treatment of spontaneous small-bowel hematoma has a good outcome in most patients. PMID- 11888456 TI - Effect of hypoventilation on bleeding during hepatic resection: a randomized controlled trial. AB - HYPOTHESIS: Blood loss in hepatic resection is an important determinant of operative outcome. OBJECTIVE: To clarify whether reducing the tidal volume would be effective in decreasing blood loss during liver transection. DESIGN: Randomized controlled trial. SETTING: University hospital. PATIENTS: Eighty patients scheduled to undergo hepatic resection were randomly assigned to receive liver transection under normoventilation (n = 40) or hypoventilation (n = 40). INTERVENTIONS: During liver transection, in the normoventilation group, the tidal volume was 10 mL/kg and the respiratory rate was 10/min; in the hypoventilation group, the tidal volume was reduced to 4 mL/kg and respiratory rate was increased to 15/min. Liver transection was performed under total or selective inflow occlusion. MAIN OUTCOME MEASURE: Blood loss. RESULTS: Between the normoventilation and hypoventilation groups, no significant difference was found in total blood loss (median [range]: 630 mL [72-3600 mL] vs 630 mL [120-3520 mL]; P =.44) or blood loss per transection area (median [range]: 7.3 mL/cm(2) [1.2 55.4 mL/cm(2)] vs 9.8 mL/cm(2) [0.9-79.9 mL/cm(2)]; P =.55). During liver transection, the central venous pressure was significantly reduced in the hypoventilation group than in the normoventilation group (median [range]: -0.7 cm H(2)O [-3.0 to 1.8 cm H(2)O] vs -0.2 cm H(2)O [-4.0 to 2.0 cm H(2)O]; P =.007). The maximum end-tidal carbon dioxide level in the hypoventilation group was significantly higher than that in the normoventilation group (maximum [range]: 50 mm Hg [28-66 mm Hg] vs 37 mm Hg [27-60 mm Hg]; P<.001). Transection time, postoperative liver function, hospitalization length, morbidity, and mortality were similar in the 2 groups. CONCLUSION: This randomized trial suggested no beneficial effect of reduction of tidal volume on bleeding during hepatic resection. PMID- 11888457 TI - St John's Wort supplements endanger the success of organ transplantation. AB - HYPOTHESIS: St John's wort is one of the most popular herbal medicines, and health care professionals often are unaware that their patients take such supplements. St John's wort causes a decrease in cyclosporine levels, thus endangering the success of organ transplantations. DESIGN: Systematic review. METHODS: Five independent computerized literature searches were conducted to identify all reports of such interactions. Data were extracted and are summarized in narrative form. RESULTS: Eleven case reports and 2 case series were located. In most instances, causality between St John's wort and the clinical or biochemical result is well established. The mechanism of interaction between St John's wort and cyclosporine has been recently elucidated and involves both P glycoprotein and cytochrome P 450 3A4 expression. Collectively these data leave little doubt that St John's wort interacts with cyclosporine, causing a decrease of cyclosporine blood levels and leading in several cases to transplant rejection. CONCLUSIONS: St John's wort can endanger the success of organ transplantations. Adequate information may be the best way to avoid future incidences. PMID- 11888459 TI - Partial matrix excision or segmental phenolization for ingrowing toenails. AB - OBJECTIVE: To decide whether partial nail extraction with phenolisation or with partial excision of the matrix should be the standard treatment in patients with ingrowing toenails of the hallux. DESIGN: Randomized clinical trial with 12-month follow-up evaluations performed by observers who did not know which procedure was applied. SETTING: Outpatient department of a surgical teaching hospital. PATIENTS: Fifty-eight consecutive patients with a total of 63 ingrowing toenails were randomized. INTERVENTION: Thirty-four partial matrix excisions ("matrix" group) and 29 phenolizations ("phenol" group) were performed. MAIN OUTCOME MEASURES: Recurrence rate, postoperative morbidity (pain, wound exudates, and scar discomfort), and time to complete recovery (wearing shoes, performing normal activities/work). RESULTS: Recurrences were seen after 7 procedures in the matrix group and also after 7 procedures in the phenol group, of which patients were symptomatic and required a second operation in 4 and 3 instances, respectively. None of the observed differences in wound healing, postoperative pain, and recovery were statistically significant. CONCLUSIONS: Partial matrix excision and phenolization are equally effective in treating ingrowing toenails. Because the use of the toxic agent phenol should be avoided, partial matrix excision is the preferable procedure. But in view of the high recurrence rate, there is a need for further improvement of the treatment of ingrowing toenails. PMID- 11888460 TI - A polymeric sealant inhibits anastomotic suture hole bleeding more rapidly than gelfoam/thrombin: results of a randomized controlled trial. AB - HYPOTHESIS: An experimental polymeric sealant (CoSeal [Cohesion Technologies, Palo Alto, Calif]) provides equivalent anastomotic sealing to Gelfoam (Upjohn, Kalamazoo, Mich)/thrombin during surgical placement of prosthetic vascular grafts. DESIGN: Randomized controlled trial. SETTING: Nine university-affiliated medical centers. PATIENTS: One hundred forty-eight patients scheduled for implantation of polytetrafluoroethylene grafts, mainly for infrainguinal revascularization procedures or the creation of dialysis access shunts, who were treated randomly with either an experimental intervention (n = 74) or control (n = 74). INTERVENTION: Following polytetrafluoroethylene graft placement, anastomotic suture hole bleeding was treated intraoperatively in all control subjects with Gelfoam/thrombin. Subjects in the experimental group had the polymeric sealant applied directly to the suture lines without concomitant manual compression. MAIN OUTCOME MEASURES: Primary treatment success was defined as the proportion of subjects in each group that achieved complete anastomotic sealing within 10 minutes. The proportion of subjects that achieved immediate sealing and the time required to fully inhibit suture hole bleeding also were compared between treatment groups. RESULTS: Overall 10-minute sealing success was equivalent (86% vs 80%; P =.29) between experimental and control subjects, respectively. However, subjects treated with CoSeal achieved immediate anastomotic sealing at more than twice the rate of subjects treated with Gelfoam/thrombin (47% vs 20%; P<.001). Consequently, the median time needed to inhibit bleeding in control subjects was more than 10 times longer than for experimental subjects (16.5 seconds vs 189.0 seconds; P =.01). Strikingly similar findings for all comparisons were observed separately for subgroups of subjects having infrainguinal bypass grafting and for those undergoing placement of dialysis access shunts. CONCLUSIONS: The experimental sealant offers equivalent anastomotic sealing performance compared with Gelfoam/thrombin, but it provides this desired effect in a significantly more rapid time frame. PMID- 11888462 TI - Inhibition of intimal hyperplasia using the selective estrogen receptor modulator raloxifene. AB - HYPOTHESIS: The selective estrogen receptor modulator raloxifene hydrochloride inhibits intimal hyperplasia. DESIGN: Controlled laboratory experiment. SETTING: Experimental animal model. INTERVENTION AND MAIN OUTCOME MEASURES: Forty-three senile female sheep were randomized to sham operation, ovariectomy, or ovariectomy followed by treatment with 17beta-estradiol or raloxifene. Six months after initial operation, we performed necropsy with histological assessment of the aortic bifurcation and bone mineral densitometry and assayed serum lipids. RESULTS: After 6 months, serum triglyceride and total and high-density lipoprotein cholesterol levels were similar among groups and within the reference range. Ovarian ablation alone resulted in intimal hyperplasia, which was attenuated with estradiol and raloxifene therapy. Furthermore, estradiol and raloxifene therapy reversed ovariectomy-induced decreases in bone mineral density measured using spine morphometry. CONCLUSIONS: Raloxifene attenuates ovariectomy induced aortic intimal hyperplasia independent of serum lipid levels. These data suggest that raloxifene, in addition to its beneficial influence on bone density, has direct, beneficial cardiovascular effects. PMID- 11888463 TI - Stapled vs excision hemorrhoidectomy: long-term results of a prospective randomized trial. AB - HYPOTHESIS: Stapled hemorrhoidectomy offers several advantages over excision hemorrhoidectomy, including reduced postoperative pain, a reduced hospital stay, and an earlier recovery time. Furthermore, stapled hemorrhoidectomy is associated with lower hemorrhoidal recurrence on long-term follow-up. DESIGN: A randomized prospective trial. Patients were blinded to the operation technique used. Follow up occurred at 1 and 3 weeks and 12 months postoperatively. SETTING: A university hospital providing primary, secondary, and tertiary care. PATIENTS: Forty patients with second- and third-degree hemorrhoid disease were randomized to undergo either stapled or excision hemorrhoidectomy. Two patients were excluded. All patients were subject to a follow-up examination. INTERVENTIONS: Stapled hemorrhoidectomy (Longo technique) vs excision hemorrhoidectomy (Ferguson technique). MAIN OUTCOME MEASURES: Operating time, postoperative pain (measured by the visual analog scale), hospital stay, histologic features, morbidity, defecation habit, continence, recovery time (return to work), and hemorrhoid recurrence at 1 year. RESULTS: Stapled vs excision hemorrhoidectomy was associated with a significantly reduced operating time (30 vs 43.25 minutes; P<.001), reduced postoperative pain scores (visual analog score) on the first 4 postoperative days (day 1: 2.7 vs 6.3; day 2: 1.7 vs 6.3; day 3: 0.8 vs 5.4; and day 4: 0.5 vs 4.8, where 0 indicates no pain, and 10, maximum pain; P < or = .001), and an earlier return to work (6.7 vs 20.7 days;P =.001). There were no differences for stapled vs excision hemorrhoidectomy in length of hospital stay (2.4 vs 2.1 days), complications (3 [15%] of 20 patients vs 5 [25%] of 20 patients), and recurrence rate (1 [5%] of 20 patients vs 1 [5%] of 20 patients). CONCLUSIONS: Stapled hemorrhoidectomy is associated with reduced postoperative pain, earlier recovery time and return to work, and a similar recurrence rate compared with the excision technique. Provided further clinical trials confirm these findings, stapled hemorrhoidectomy may become a future gold standard. PMID- 11888464 TI - Intraoperative parathyroid hormone measurement in patients with secondary hyperparathyroidism. AB - HYPOTHESIS: Secondary hyperparathyroidism decreases renal clearance of parathyroid hormone (PTH). OBJECTIVE: To determine whether rapid PTH assays can be used to predict the success of a total parathyroidectomy to treat symptomatic secondary hyperparathyroidism. DESIGN: Case series from August 1 to December 31, 2000. SETTING: Tertiary referral center. PARTICIPANTS: Patients with symptomatic secondary hyperparathyroidism (n = 24) who underwent total parathyroidectomy and autotransplantation were included in the study. INTERVENTIONS: Blood samples for rapid PTH analyses were drawn from an indwelling catheter at the induction of anesthesia (baseline) and before (0 minutes), 10 minutes, and 30 minutes after the removal of the last parathyroid gland. Regular intact PTH (iPTH) assays were conducted later. MAIN OUTCOME MEASURE: If a patient's regular iPTH levels were below 65 pg/mL at 1 week or 3 months postoperatively, the operation was considered successful. RESULTS: All 24 patients had successful operations. Rapid PTH and regular iPTH correlated significantly at 0, 10, and 30 minutes. Rapid PTH levels decreased significantly at each time period and were 176 +/- 40.9 pg/mL (mean +/- SE) at 10 minutes. The percentage decrease in rapid PTH levels was 39.5% +/- 12.7% at 0 minutes, 75.1% +/- 6.2% at 10 minutes, and 91.0% +/- 0.1% at 30 minutes (mean +/- SE). A decrease of 60% or more from baseline PTH levels at 10 minutes and/or a decrease of 85% or more at 30 minutes predicted the successful removal of all parathyroid glands. CONCLUSIONS: A drop in PTH levels is delayed until 30 minutes after total parathyroidectomy; however, a rapid PTH assay 10 minutes after the removal of the last parathyroid gland is as accurate as an assay performed at 30 minutes postoperatively. Intraoperative PTH monitoring demonstrates relevant decreases in rapid PTH levels after parathyroidectomy that are similar to those previously documented in patients with primary hyperparathyroidism. PMID- 11888465 TI - A logical and stepwise operative approach to radical neck dissection. AB - A thorough understanding of the anatomy of the neck is essential to avoid injury to vital structures when performing radical neck dissection. The complicated anatomical relations of the various nerves, vessels, and muscles within the confined area of the neck can often be daunting. We outline strategic anatomical landmarks and their relationships to important nerves, arteries, veins, and lymphatics to simplify the complicated and formidable anatomy of the neck. We also delineate key maneuvers that, when combined with the anatomical landmarks, offer a stepwise and logical approach to performing radical neck dissection that confers improved safety. PMID- 11888466 TI - Surgery in Brazil. PMID- 11888467 TI - Image of the month. Pectus excavatum. PMID- 11888468 TI - Surgical reminiscence: on call. PMID- 11888469 TI - Moments in surgical history: Arthur Emanuel Hertzler (1870-1946). PMID- 11888470 TI - Outcomes of extended lymph node dissection for squamous cell carcinoma of the thoracic esophagus. AB - Patients with thoracic esophageal carcinoma who underwent extended lymph node (LN) dissection were studied to assess the state of LN metastasis and evaluate its outcome in terms of a prognostic benefit. Pertaining to LN metastasis, it was found that depending on the location of a primary tumor, the area of choice, in which metastasis tends to develop predominantly, showed some variation. However, irrespective of the location of the tumor, the predominant growth of positive nodes was found to locate among three fields, namely the neck, mediastinum and abdomen even in patients with a single metastatic node. This suggests that extended LN dissection including the neck, mediastinum and abdomen should be considered mandatory, if a complete removal of the tumors for carcinoma of the thoracic esophagus is to be desired. Multivariate analysis revealed importance of LN dissection as a prognostic factor. A cumulative survival rate in the patients with lymphadenectomy through right thoracotomy was statistically better than that in the patients who underwent blunt extraction of the esophagus without lymphadenectomy. Furthermore, extensiveness of LN dissection could effectively serve as a prognostic factor. Consequently, three-field LN dissection yields a prognostic benefit to improve a long term survival in patients with carcinoma of the thoracic esophagus. PMID- 11888472 TI - Growth of the pulmonary arteries after systemic-pulmonary shunt. AB - Pulmonary artery growth after a systemic-pulmonary shunt was angiographically evaluated in 19 out of 35 patients. The mean age of the subjects at the time of the initial operation was 18+/-18 months including 12 patients under a year old. The preoperative diagnosis was tetralogy of Fallot (TOF) in 10 patients, TOF plus pulmonary atresia in five and transposition of great arteries in four. A Blalock Taussig shunt (BTS) operation was performed in 16 patients (15 classical and 1 modified) and a central shunt was performed in three patients as an initial operation. The preoperative pulmonary artery index (PAI) was 129+/-42 in all patients and there were no significant differences between patients under or over a year old (139+/-42 vs. 115+/-49). Postoperative angiography was performed 32+/ 13 months after the surgery. Room air arterial O2 pressure increased significantly from 29+/-5 mmHg to 42+/-5 mmHg just after an initial palliative shunt operation. PAI change in patients under a year old was 214+/-73%, which was higher than 145+/-27% in patients over a year old after a palliative shunt operation. On the ipsilateral side, PAI change was almost the same between patients under and over a year old. On the contralateral side, PAI change in patients under a year old was 216+/-68%, which was significantly higher than the 116+/-21% in patients over one year old. There was a significant negative correlation (r=-0.65, p<0.05) between PAI change and arterial O2 pressure as measured just after a palliative shunt operation. In conclusion, a palliative shunt operation prior to a year old is desirable in order to produce sufficient and bilateral pulmonary artery growth. PMID- 11888471 TI - Bronchopleural fistula in the surgery of non-small cell lung cancer: incidence, risk factors, and management. AB - BACKGROUND: The incidence of a bronchopleural fistula (BPF) as a major complication after non-small cell lung carcinoma (NSCLC) surgery has decreased in recent years, due to new surgical refinements and a better understanding of the bronchial healing process. We reviewed our most recent experience with BPFs and tried to determine methods which may effectively reduce its occurrence. METHODS: Data on 490 patients with lung resections for NSCLC over a period from 1990 to 1999 were retrospectively reviewed. Details regarding surgery and the subsequent treatment were carefully reviewed. Particular attention was paid to factors possibly affecting the occurrence of BPFs: the technique of the initial bronchial closure, previous radiation and/or chemotherapy, need for postoperative ventilation and presence of residual carcinomatous tissue at the bronchial suture line. Information about age, sex, clinical diagnosis, associated conditions, TNM stage, period between primary operation and rethoracotomy and postoperative outcome was also recorded. RESULTS: The overall BPF incidence was 4.4% (22/490). There were 21 (95.5%) males and 1 (4.5%) female, mean age was 57.8 years. BPFs occurred after pneumonectomy in 12 (54.6%), after lobectomy in 9 (40.9%) patients and after sleeve resections in 1 (4.5%) patient. Mortality rate was 27.2% (6/22). Right-sided pneumonectomy and postoperative mechanical ventilation were identified as risk factors for BPFs (p<0.05). Initial chest re-exploration was performed in 20 (90.9%) patients. After debridement, the bronchial stump was reclosed by hand suture in 10 (45.4%) patients. All 10 (45.4%) patients with a post-lobectomy- and sleeve resection BPF necessitated completion surgery. The BPF was additionally covered with a vascularized flap in 20 (90.9%) patients. In 2 (9%) patients with small BPFs and poor overall condition the initial treatment was endoscopic. In both the fistula persisted and the stump had to be surgically resutured. CONCLUSIONS: A BPF remains a major complication in the surgery of NSCLC because of its high mortality and morbidity rate. A BPF is more common after right-sided pneumonectomy and is frequently associated with postoperative mechanical ventilation. The management varies according to the initial type of surgery, the size of the BPF, the overall patient condition and that of the remaining lung. Endoscopic treatment is reserved only for small fistulas associated with poor general condition. PMID- 11888473 TI - Resuscitation and evaluation of non-beating hearts obtained from asphyxiated dogs via autoperfusing heart-lung circuit. AB - BACKGROUND: The shortage of donor hearts has made use of non-beating hearts as cardiac grafts an attractive possibility for heart transplant candidates. The purpose of this study was to evaluate the utility of leukocyte-depleted hot shot cardioplegia for resuscitation of non-beating hearts obtained from asphyxiated dogs via an autoperfusing heart-lung circuit. METHODS: Mongrel dogs were divided into 3 groups according to the warm ischemia time and the method of reperfusion before starting the autoperfusing heart-lung circuit. Group A (n=4) had 60 minutes of warm ischemia and reperfusion without leukocyte-depleted hot shot, Group B (n=5) had 30 minutes of warm ischemia and reperfusion with leukocyte depleted hot shot, and Group C (n=7) had 60 minutes of warm ischemia and reperfusion with leukocyte-depleted hot shot. We calculated stroke work via the heart-lung circuit to evaluate cardiac function of the resuscitated hearts. The criteria for "recovery" has been reported elsewhere. Myocardial water content of the resuscitated hearts was also measured and analyzed. No inotropic agents were used. RESULTS: The recovery rates in groups A, B and C were 0%, 80% and 57%, respectively, and the group B rate was significantly higher than the group A rate (p=0.04). Although myocardial water content did not differ between groups B and C, it was significantly lower in recovered hearts than in non-recovered hearts (p=0.04). Significant negative correlation was observed between the maximum stroke work value and myocardial water content in the resuscitated hearts (r=0.668, p=0.03). CONCLUSIONS: The autoperfusing heart-lung circuit is useful for evaluation and maintenance of cardiac function. Our experimental data shows that leukocyte-depleted hot shot plays a great role for resuscitation and recovery of non-beating hearts. PMID- 11888474 TI - Angiographic assessment of surgical reconstructions for left ventricular asynergy with indices of abnormally contracting segments. AB - BACKGROUND: In surgical reconstruction for left ventricular asynergy after myocardial infarction, the conventional linear closure technique second to simple resection and endoventricular circular patch plasty, which is the so-called Dor technique, are commonly utilized. We assessed these techniques using an abnormally contracting segment (ACS) in the left ventriculogram. METHODS: We reviewed 10 and 15 patients who underwent the linear technique (group L) and the Dor technique (group D), respectively. %ACS was determined as the percent ratio of both akinetic and dyskinetic chords among the total chords in the centerline method of regional wall motion analysis. A difference between preoperative and postoperative ejection fraction (EF) was generated by preoperative EF and this percentage ratio was determined as %EF. RESULTS: Postoperative EF improved from 31% to 41% in group L and from 33% to 49% in group D (p<0.05). Postoperative EF in group D was higher than in group L (p<0.05). %ACS decreased from 41% to 34% in group L and from 41% to 19% in group D (p<0.05). Postoperative %ACS was lower in group D than in group L (p<0.05). The significant correlation between preoperative %ACS and %EF was negative in group L and positive in group D (p<0.05). CONCLUSIONS: The Dor technique is more effective for the postoperative systolic function than the linear technique because more extensive reduction in %ACS is possible with the Dor technique than with the linear technique. Dor technique becomes more crucial to the postoperative systolic function as the preoperative %ACS becomes larger. PMID- 11888475 TI - Re-exploration for hemorrhage following open heart surgery differentiation on the causes of bleeding and the impact on patient outcomes. AB - OBJECTIVES: To differentiate surgical bleeding requiring re-exploration from postoperative coagulopathy and determine the differences in patient outcomes. METHODS: This was a retrospective chart review of 2,263 adult patients undergoing elective and emergency open heart procedures encompassing coronary artery bypass, valvular, and a combined procedure to determine the impact of source of bleeding leading to re-exploration. RESULTS: Eighty-two patients (3.6%) required re exploration. Sixty-six percent had surgical bleeding; the remaining 34% were coagulopathic. Postoperative coagulopathy was associated with preoperative heparin use (37% vs. 19.9% for controls p<0.05). Re-operative procedures combined bypass/ valve (p<0.001) and prolonged cardiopulmonary bypass and aortic cross clamp times (p<0.05) were more prevalent in the coagulopathy group. Postoperative inotrope use was increased in patients who were re-explored (p<0.001), as were cardiac, pulmonary, renal and abdominal complications (p<0.001), and in all cases those patients with medically related bleeding had worse acute outcomes than the group with surgical causes for re-exploration. The hospital stay was prolonged for both patients with surgical bleeding (23.5 days) and patients with coagulopathy (27.1 days) compared to patients not undergoing re-exploration for bleeding (12.0 days, p<0.001). Survival was 91.3% for patients with surgical bleeding, 87.5% for patients with coagulopathy, and 98.0% for all others (p<0.01). CONCLUSIONS: Severe postoperative hemorrhage is associated with significant morbidity and increased mortality. Postoperative hospital stay, morbidity, and mortality were significantly worse in patients suffering from coagulopathy when compared to those patients with hemorrhage from surgical causes. PMID- 11888476 TI - Management of the heart rate during coronary artery bypass grafting on the beating heart: newly devised methods of decreasing heart rate--a preliminary report-. AB - BACKGROUND: To develop new methods for achieving bradycardia, we studied the feasibility of producing transient, reversible bradycardia with atrial stimulation and cooling of the sinoatrial node. METHODS: In an animal study, the atrium was stimulated electrically during the refractory period of the atrioventricular node. Alternatively, an area of the sinoatrial node was cooled regionally. The two methods were also performed in combination. In a clinical study, atrial stimulation was applied in seven consecutive patients who underwent coronary artery bypass grafting (CABG). RESULTS: In the animal study, atrial stimulation was effective only when 2 mg/kg of diltiazem was administered. Such atrial stimulation decreased heart rate (beats/minute) from 95.8+/-16.9 to 64.2+/ 20.0 (the average reduction from the control value 66.1+/-10.3%). Cooling the sinoatrial node decreased heart rate, and was effective with or without administration of diltiazem. Heart rate was decreased from 156.6 31.7 to 110.7+/ 21.7 (average reduction from control value 71.3+/-9.2%) before using diltiazem and from 102.0+/-11.9 to 63.5+/-13.9 (average reduction from control value 62.0+/ 10.4%) after administration of diltiazem. By combining the two methods, heart rate was decreased from 102.0+/-12.3 to 44.6+/-9.1 (average reduction from control value 43.5+/-6.3%). In our clinical study, the atrial stimulation method was effective. CONCLUSION: Atrial stimulation or regional cooling of the sinoatrial node slowed the heart rate. By combining the two methods, the heart rate was slowed to 40. Clinically, atrial stimulation was effective in CABG patients. PMID- 11888477 TI - A case of primary leiomyosarcoma of the chest wall successfully resected under the video-assisted thoracoscopic approach. AB - We report a case of a 62-year-old woman with primary leiomyosarcoma of the chest wall which was successfully resected under the video-assisted thoracoscopic approach. The disease was found during the treatment for a malignant melanoma of the left heel. On the preoperative CT images, the lesion was suspected to be a metastasis of the malignant melanoma. The thoracoscopic surgery revealed that the tumor originated from the parietal pleura, and it was resected with a 5-mm margin of normal pleura. Histopathologically, the tumor was diagnosed as low-grade leiomyosarcoma. Since no residual tumor cells were proven in the resected margins histologically, further resection was not performed. At present, she is alive and well with no sign of recurrence of leiomyosarcoma two years and one month after operation. Thoracoscopic surgery is worth trying for accurate diagnosis of and effective treatment for a chest lesion without apparent invasion of the chest wall on the preoperative CT images. PMID- 11888478 TI - Post radiation inflammatory malignant fibrous histiocytoma arising from the chest wall. AB - A 59-year-old man who underwent radiation therapy (41 Gy) to the mediastinum through the anterior chest for Hodgkin's disease presented with a painful anterior chest wall tumor 18 years later. The tumor originated from the left parasternal region and was excised with the sternum. Chest wall reconstruction was performed. The tumor measured 45 x 45 mm and invaded the sternum. The pathologic diagnosis was malignant fibrous histiocytoma. Early and complete excision of the tumor is indicated. PMID- 11888479 TI - Open heart operation in a patient with hereditary spherocytosis: a case report. AB - A 9-year-old girl who had ostium secundum atrial septal defect (ASD) and hereditary spherocytosis (HS) is described. The patient had a history of splenectomy for HS and underwent repair of the ASD under cardiopulmonary bypass (CPB), however, no significant or persistent hemolysis was observed during and after CPB. Only 10 patients with HS who underwent cardiac operations using CPB have been reported. The case is presented due to its rarity. PMID- 11888480 TI - Aortic valve replacement for aortic stenosis with a small aortic annulus in a patient having Werner's syndrome and liver cirrhosis. AB - Werner's syndrome is a rare genetic disease characterized by premature aging and scleroderma-like involvement of the skin. We report a case of aortic valve replacement for severely calcified aortic valve stenosis with a small annulus in a patient suffering from Werner's syndrome and liver cirrhosis PMID- 11888481 TI - Simultaneous mitral valve replacement and bypass grafting for mycotic aneurysm of the femoral artery during the active phase of infective endocarditis: a case report. AB - A 52-year-old woman with a 3-week history of fever and cough was diagnosed as having bacterial endocarditis with vegetation and severe mitral valve insufficiency by echocardiography. Blood culture revealed Streptococcus mitis. After antibiotic treatment for 3 weeks, the patient noticed swelling with pain in her left groin. Computed tomography revealed an occluded aneurysm in the left common femoral artery. Simultaneous surgical treatments of mitral valve replacement and bypass grafting using a saphenous vein following resection of the mycotic femoral arterial aneurysm were performed. Pathohistological examination of surgical specimens revealed acute inflammatory findings, but no microorganisms were found, probably because of the preoperative antibiotic therapy. Her postoperative course was uneventful, and there was no recurrence of mycotic aneurysms in a period of 10 months after the operation. Prompt recognition and urgent simultaneous surgical treatments for mycotic aneurysms complicated with infective endocarditis were effective. PMID- 11888482 TI - Extended aortic grafting for acute ascending dissection after type B dissection. AB - A 31-year-old man was diagnosed with acute ascending aortic dissection and massive aortic regurgitation following acute type B dissection during drug treatment. Although the aortic arch was not dissected, we performed aortic replacement from the aortic root to the proximal portion of the descending aorta. The aim of the operation was the prevention of aortic arch dissection, and closure of initial entry of type B dissection. PMID- 11888483 TI - Arterial wall properties and Womersley flow in Fabry disease. AB - BACKGROUND: Fabry disease is an X-linked recessive lysosomal storage disease resulting in the cellular accumulation of globotriaosylceramide particularly globotriaosylceramide. The disease is characterized by a dilated vasculopathy with arterial ectasia in muscular arteries and arterioles. Previous venous plethysomographic studies suggest enhanced endothelium-dependent vasodilation in Fabry disease indicating a functional abnormality of resistance vessels. METHODS: We examined the mechanical properties of the radial artery in Fabry disease, a typical fibro-muscular artery. Eight control subjects and seven patients with Fabry disease had a right brachial arterial line placed allowing real time recording of intra-arterial blood pressure. Real time B-mode ultrasound recordings of the right radial artery were obtained simultaneously allowing calculation of the vessel wall internal and external diameter, the incremental Young's modulus and arterial wall thickness. By simultaneously measurement of the distal index finger-pulse oximetry the pulse wave speed was calculated. From the wave speed and the internal radial artery diameter the volume flow was calculated by Womersley analysis following truncation of the late diastolic phase. RESULTS: No significant difference was found between Fabry patients and controls for internal or external arterial diameters, the incremental Young's modulus, the arterial wall thickness, the pulse wave speed and the basal radial artery blood flow. Further, no significant difference was found for the radial artery blood flow in response to intra-arterial acetylcholine or sodium nitroprusside. Both drugs however, elevated the mean arterial flow. CONCLUSIONS: The current study suggests that no structural or mechanical abnormality exists in the vessel wall of fibro-muscular arteries in Fabry disease. This may indicate that a functional abnormality downstream to the conductance vessels is the dominant feature in development Fabry vasculopathy. PMID- 11888485 TI - Reactions to local anesthetics. PMID- 11888484 TI - How well do second-year students learn physical diagnosis? Observational study of an Objective Structured Clinical Examination (OSCE). AB - BACKGROUND: Little is known about using the Objective Structured Clinical Examination (OSCE) in physical diagnosis courses. The purpose of this study was to describe student performance on an OSCE in a physical diagnosis course. METHODS: Cross-sectional study at Harvard Medical School, 1997-1999, for 489 second-year students. RESULTS: Average total OSCE score was 57% (range 39-75%). Among clinical skills, students scored highest on patient interaction (72%), followed by examination technique (65%), abnormality identification (62%), history-taking (60%), patient presentation (60%), physical examination knowledge (47%), and differential diagnosis (40%) (p <.0001). Among 16 OSCE stations, scores ranged from 70% for arthritis to 29% for calf pain (p <.0001). Teaching sites accounted for larger adjusted differences in station scores, up to 28%, than in skill scores (9%) (p <.0001). CONCLUSIONS: Students scored higher on interpersonal and technical skills than on interpretive or integrative skills. Station scores identified specific content that needs improved teaching. PMID- 11888486 TI - Risk factors for latex sensitization in children with spina bifida. AB - BACKGROUND: Children with spina bifida represent the major risk group for latex sensitization. PURPOSE: To determine the prevalence of latex sensitization in these children and to identify risk factors. MATERIAL AND METHODS: We studied 57 patients with spina bifida. The mean age was 5.6 years and the male/female ratio was 0.8/1. In all patients a questionnaire, skin prick test (SPT) with latex (UCB Stallergenes, Lofarma and ALK-Abello), common aeroallergens and fruits (UCB Stallergenes) and serum determination of total IgE (AlaSTAT) were performed. RESULTS: The prevalence of latex sensitization was 30 %; only two sensitized children (12 %) had symptoms after exposure. Risk factors for latex sensitization were age >/= 5 years (p = 0.008; OR = 6.0; 95 % CI = 1.7-22.1), having at least four previous surgical interventions (p < 0.0001; OR = 18.5; 95 % CI = 3.6-94.8), having undergone surgery in the first 3 months of life (p = 0.008; OR = 5.4; 95 % CI = 0.7-29.2) and total serum IgE >/= 44 IU/ml (p = 0.03; OR = 3.8; 95 %CI = 1.1 13.1). Multiple logistic regression analysis showed that only a history of four or more surgical interventions (p < 0.0001; OR = 26.3; 95 %CI = 2.9-234.2) and total serum IgE >/= 44 IU/ml (p = 0.02; OR = 8.6; 95 % CI = 1.4-53.4) were independently associated with latex sensitization. Sex, family and personal allergic history, hydrocephalus with ventriculoperitoneal shunt, cystourethrograms, intermittent bladder catheterization and atopy were not related to latex sensitization. CONCLUSIONS: In children with spina bifida, significant and independent risk factors identified for latex sensitization were multiple interventions and higher levels of total serum IgE. A prospective study will clarify the clinical evolution of assymptomatic children sensitized to latex. PMID- 11888487 TI - Allergy to local anaesthetics in dentistry. Myth or reality? AB - BACKGROUND: Local anesthetics (LA) are frequently used in dentistry. Although these drugs are usually well-tolerated, they can sometimes provoke adverse reactions of various types and severity. The true incidence of LA allergic reactions is unknown. The objectives of this study were (i) to evaluate the incidence of immediate adverse events in subjects requiring local anesthetic injection in order to receive dental treatment; (ii) to assess the incidence of anaphylactic allergic reactions among those recorded as adverse events and (iii) to analyze the relationship between the atopic antecedents of these patients and documented allergic reactions. MATERIALS AND METHODS: A prospective, open-label, non-comparative study including 5,018 subjects who received LA during dental treatment, despite their age, was carried out in 7 private or public odontological centers. All the possible reactions that could appear during the first hour of anesthetic administration were assessed. RESULTS: Twenty-five adverse reactions were diagnosed, representing 0.5 % of the study population. None of these reactions was due to an allergic cause. Most (22/25) were mild, quickly reversible psychogenic or vasovagal reactions. One case was related to defects in the anesthetic technique. In two further cases, allergic etiology was ruled out after skin and dose provocative challenge tests with the anesthetic. In conclusion, allergic reactions to LA are very rare. Most adverse reactions are psychogenic or vasovagal. Physicians and dentists should be aware of these facts in order to minimize the frequent fears and myths concerning the use of LA in the dentist's office. PMID- 11888488 TI - [Immunotherapy with acarus extract in children under the age of 5 years]. AB - BACKGROUND: Specific immunotherapy with allergens is the only treatment capacite of modifying the natural course of allergic diseases. Although its effectiveness is higher when started in early stages, WHO guidelines still consider age under 5 years as relative contraindication.A review of a group of 22 children starting immunotherapy before the age of 5 years in a pediatric allergy unit was attempted to assess the effectiveness and safety of this treatment. METHODOLOGY AND RESULTS: Clinical charts of 22 children with house dust mite asthma starting conventional immunotherapy before the age of 5 years were reviewed. In all cases, biologically standardized extracts and conventional subcutaneous schedules have been used and administration was performed in a hospital setting. Effectiveness and safety were evaluated by clinical scores. The average treatment period was 16.95 ( 10.12) months. Systemic reactions (moderate) were observed in 7 children and local reactions in 5. Children treated for more than 1 year (15 patients) showed a decrease in the number of yearly acute exacerbations (from 5.13 to 2.57) and in hospital admissions (from 5 to 2 children). In 10 patients, drug requirements for bronchial asthma were reduced. CONCLUSION: The clinical data if this study suggest that house dust mite specific immunotherapy under hospital control can be begun in young children with good tolerance and clinical improvement. PMID- 11888489 TI - Asthmatic children and risk factors at a province in the southeast of Turkey. AB - BACKGROUND: The prevalence of asthma appears to be on the increase and the risk factors are not well established. Environmental and demographic characteristics of asthmatic children were investigated to identify the risk factors accompanying asthma. METHODS: 140 asthmatic children aged at 3-15 years were compared with 96 age-matched control subjects admitted to the paediatric outpatient clinics of Dicle University Hospital. Information about the children were obtained from parents and patients'charts. RESULTS: Patients with asthma were most frequently admitted in May-June and November-January months. Association of the disease with allergic rhinitis was found in 84 children (60 %), allergic conjunctivitis in 63 (45 %), atopic dermatitis in 29 (21 %) and gastrointestinal symptoms in 18 (13 %). Mean age of the cases was 6.8 3.3 and 6.9 3.2 years (p > 0.05); male to female ratios were 91/49 and 43/53 (p = 0.002) in the study and control groups, respectively. There were significant differences at number of cases stated to have any symptoms induced by air pollution (25.6 %, 3.1 %, p < 0,001), exercise (47.5 %, 4.2 %, p < 0.001) and cold exposure (33 %, 15.6 %, p = 0.03), but not by damp, dust, indoor smoking, foods, drugs, and animal contact (p > 0.05) between the study and the control groups, respectively. Family history of atopy was 66 % and 8.4 %, (p < 0.001) in the study and control groups, respectively. Family crowding index, duration of breast feeding, parental education and number of consanguineous married parents were not different between both groups. History of upper respiratory tract infections were more frequent in asthmatic children than controls. Children with an earlier age of onset ( 5 years. CONCLUSIONS: Many risk factors, both individual and environmental are associated with asthmatic children in Diyarbakr. Among many risk factors that aggrevating asthma in children in Diyarbakr, air pollution, cold exposure and upper respiratory infections are preventable. PMID- 11888490 TI - Asthma and respiratory disease mortality rates in the state of Sao Paulo, Brazil: 1970-1996. AB - BACKGROUND: Asthma morbidity and mortality have been increasing. Data of asthma and respiratory mortality rates in Brazil are scarse. METHODS: We studied asthma and respiratory disease mortality rates in the state of So Paulo (capital and country side) from 1970 to 1996, as its relation with sales of drugs usually used in asthma treatment. RESULTS: Asthma mortality in the 5-34-year age group has doubled in the state of So Paulo (0.2 deaths/100,000 inhabitants in 1970 to 0.4 in 1996), mainly by its increase in the capital. The greatest increase was observed in the population of up to 15 years of age. The sales of inhaled anti inflammatory drugs are proportionally very low and reflect a greater concern about the treatment of acute exacerbations. CONCLUSION: We believe that the institution of a public health supply to the whole population could provide better conditions for the control of those indexes. PMID- 11888492 TI - [Common variable immunodeficiency in children]. AB - Common variable immunodeficiency (CVID) is one of the more frequent primary immunodeficiencies (PID), after IgA deficiency, and affects a heterogeneous group of patients of various ages and with autosomal recessive inheritance. Our objective is to present the group of children diagnosed with CVID treated in our Hospital Infantil Vall d'Hebron and comment on the diagnostic problems that can arise. Sixteen boys and girls were diagnosed between the ages of 7 months and 15 years. The diagnosis is based on low immunoglobulins and a clinical picture of infection. Differential diagnosis in the paediatric age must consider mainly other PIDs: transient hypogammaglobulinaemia of infancy, X chromosome-linked agammaglobulinaemia (XLA), X chromosome-linked hyper IgM syndrome (X-HIM), IgG subclass deficiency and IgA deficiency (IgAD). Other processes that evolve with recurrent respiratory infections, such as cystic fibrosis, must also be discarded. CONCLUSIONS: These patients present a high incidence of respiratory infections and bronchiectasias. We also observe associated allergic and autoimmune processes. Early diagnosis is indispensable to initiate suitable treatment and avoid the consequences of both respiratory and digestive infections. PMID- 11888491 TI - An extensively hydrolysed cow's milk formula improves clinical symptoms of gastroesophageal reflux and reduces the gastric emptying time in infants. AB - BACKGROUND: About 20 % of infants fed with breast-milk substitutes suffer from Gastro Esophageal Reflux (GER) and 1/3 of them also show Cow's Milk Allergy (CMA) symptoms. METHODS: We planned this study to assess by dynamic echography the usefulness of an Extensively Hydrolysed Cow's Milk Formula (eHF) in infants suffering from GER. Ten infants showing GER symptoms and 10 normal babies, all fed with breast-milk substitutes, were enrolled. Clinical symptom scores related to GER were assessed for one week. The Gastric Emptying Time (GET) was determined by means of dynamic echography after feeding with cow's milk-derived formulae and again after a week feeding with eHF in subjects previously showing GER symptoms. RESULTS: All infants with a clinical diagnosis for GER showed an abnormally high average GET in comparison to normal subjects (205 vs 124 min, p = 0.000). Switching to the eHF led to a significant clinical improvement (p = 0.0039) especially in babies skin-test and RAST positive to cow's milk, and to a significant decrease toward the normal value of the GET (167 min, p < 0.001). CONCLUSIONS: The eHF tested improves GER symptoms in infants suffering from this disease. Our experience confirms and supports the use of dynamic echography as a reliable, simple, and non-invasive diagnostic method for infants with an increased GET associated with clinical symptoms of GER. PMID- 11888493 TI - Facial edema associated with thyroid autoimmunity. AB - BACKGROUND: There is growing evidence that some cases of chronic urticaria and angioedema are associated with thyroid autoimmunity. We present the case of a 42 year old woman with a two month long evolution of edema in the right hemiface associated with thyroid antibodies. METHODS: Skin tests were performed with a standard series of aeroallergens and foods and were negative. Epicutaneous tests were performed with a series of contact allergens with a negative result. The chest X-ray was normal. A complete blood count was performed with ESR, biochemistry study, proteinogram, coagulation, complement, rheumatoid factor, anti-nuclear antibodies, hydatidosis and hepatitis serologies, urine sediment and stool parasitology, which were normal. The TSH was normal and, among the thyroid antibodies, the thyroglobulin ones (492 IU/mL) were positive and the microsomal ones were negative. RESULTS: The edema remitted with the 2-month long thyroxin treatment. In spite of the clinical response, and although the thyroglobulin antibodies initially suffered a small decrease, they increased to values which greatly surpassed the initial ones, coinciding with a relapse of the facial edema, so that treatment was re-initiated with thyroxin, and the picture subsided again. CONCLUSIONS: Some cases of localized edema can be associated with thyroid antibodies and respond to treatment with thyroxin. PMID- 11888494 TI - [Comparative study of the type of obesity in pre- and postmenopausal women: Relationship with fat cell data, fatty acid composition and endocrine, metabolic, nutritional and psychological variables]. AB - BACKGROUND: The weight increase that many women experience during menopause may be the result of ageing. However, the precise factors inducing obesity during this period remain to be identified. The object of this study was to determine the type of obesity in a group of women along with its distinctive features, if any, as a function of the menopause stage. PATIENTS AND METHOD: The sample consisted of 55 women (22 premenopausal and 33 postmenopausal) with grade I and II obesity. Distribution of body fat, composition of the adipose tissue, size and number of adipocytes, lipidic and hormonal profile as well as nutritional and psychological aspects were all taken into account. RESULTS: Postmenopausal women had an android distribution of fat, whereas it was gynoid in premenopausal women. The adipose tissue showed different cell characteristics, the number of fat cells and content of saturated fatty acids (myristic and palmitic) being significantly lower in the postmenopausal group. Menopause was associated with an increase in plasmatic lipids and a decrease in the levels of certain hormones (dehydroepiandrosterone-sulphate and insulin). Postmenopausal women tended to have healthier eating habits than premenopausal women, with a significantly lower fat intake but higher carbohydrate and fibre intakes. However, the degree of physical activity was lower than in premenopausal women. CONCLUSIONS: The type of obesity differs as a function of the menopausal status, a finding that should be taken into account when establishing a dietetic treatment. PMID- 11888496 TI - [Detection by nested-PCR of Toxoplasma gondii in patients infected with human immunodeficiency virus]. AB - BACKGROUND: Our purpose was to evaluate the effectiveness of the nested polymerase chain reaction (nested-PCR) for the laboratory diagnosis of active toxoplasmosis in patients infected with human immunodeficiency virus (HIV). PATIENTS AND METHOD: The study was carried out in 157 HIV-infected patients. We determined IgG, IgM and IgA antibodies against Toxoplasma gondii as well as parasite DNA by nested-PCR in blood samples. RESULTS: IgG antibodies were detected in 56 patients, IgM in 1 and IgA in 5 patients. Eleven patients were found to have DNA of the parasite. CONCLUSIONS: The polymerase chain reaction is a rapid, sensitive and effective technique in the early diagnosis of toxoplasmosis in HIV-infected patients. Its positivity may also point to the beginning of treatment in asymptomatic individuals. PMID- 11888497 TI - [Renal function protection in type 2 diabetic patients]. PMID- 11888495 TI - [Significant incidence of type 2 diabetes on high-risk Spanish population. The IGT Study (2)]. AB - BACKGROUND: Our purpose was to estimate the incidence of type 2 diabetes among a high risk population with or without impaired glucose tolerance (IGT), analysing the progression to diabetes. PATIENTS AND METHOD: Multicenter cohort study on high risk individuals without diabetes (WHO-85 criteria) in primary health care. Subjects underwent an oral glucose tolerance test measuring fasting plasma glucose (FPG) and plasma glucose at 2 hours (2hPG). Mean follow-up was 37.2 months (4.3-69.7). Phenotypic features, diagnostic variation, diabetes incidence and predictive factors (multivariate analysis and Cox proportional hazards model) were assessed. RESULTS: We included 243 individuals (148 females), aged 59.7 (10) years, with FPG < 7.8 mM and 2hPG < 11.1 mM. 137 IGT subjects (56.4%) and 106 (43.6%) normal glucose tolerance (NGT) subjects with a similar risk factor impact were evaluated. After the study was closed, 63 (25.9%) subjects developed diabetes: 43 (31.4%) with baseline IGT and 20 (18.9%) with NGT. Overall diabetes incidence increased over time but not proportionally. Mean annual incidence was 9.2% and it dropped to 4.6% when FPG was used as the unique diagnosis criterion (ADA-97). Male under 65 years with both overweight and IGT reported the highest incidence. HbA1c, FPG and 2hPG were independent predictors. Increased HDL cholesterol showed a protective effect on diabetes incidence. The IGT diagnosis interval was a much better predictor [OR = 2.06 (1.56-2.72)] of diabetes development than the impaired FPG diagnosis interval [OR = 1.37 (0.93-2.04)]. CONCLUSIONS: FPG predicted but undervalued diabetes incidence in high risk population. The IGT (2hPG) diagnosis interval predicted diabetes development better than the impaired fasting plasma glucose diagnosis interval. Increased diabetes incidence in high risk Spanish population, particularly with regard to IGT, means that primary preventive resources should be increased. PMID- 11888498 TI - [Life-threatening hemoptysis in cystic fibrosis: clinical characteristics and management in 36 episodes]. AB - BACKGROUND: Our goal was to evaluate the clinical characteristics and management of life-threatening hemoptysis in patients with cystic fibrosis (CF). PATIENTS AND METHOD: We included adult CF patients followed up at the Cystic Fibrosis Units of the Autonomous Community of Madrid who had life-threatening hemoptysis from June 1990 to December 1999. RESULTS: Twelve CF patients (4 females) developed 36 episodes of life-threatening hemoptysis (30 massive and 6 recurrent). Lung disease was moderate to severe. Sputum cultures revealed Pseudomonas aeruginosa in 10 patients. Thirteen episodes (36%) resolved upon antibiotic treatment and 3 (8%) after antibiotic therapy and bronchoscopy. Bronchial artery embolization (BAE) was performed in 20 of 36 events. Immediate technique success was achieved in 80% episodes (16 of 20) after one session, 85% (17/20) after two sessions, and 95% (19/20) after three sessions. No major complications associated with the procedure were seen. The overall recurrence rate per episode was 69% (24 of 35 episodes in 6 patients) with a mean time of recurrence of 13 months. There were no massive hemoptysis-associated deaths during the follow-up. CONCLUSIONS: Life-threatening hemoptysis is a frequent complication in CF patients who have moderate or severe lung disease. When conservative therapeutic measures (including antibiotics) fail to control it, BAE should be performed. When performed by expert professionals, BAE is effective and safe to immediate control of life-threatening hemoptysis in patients with CF. PMID- 11888499 TI - [Unique or high resolution consultation as an efficiency alternative to conventional hospital outpatients consultations]. PMID- 11888500 TI - [Cardiovascular disease in diabetes mellitus]. PMID- 11888501 TI - [Diabetic nephropathy]. PMID- 11888502 TI - [Fatal cytomegalovirus infection in a patient after splenectomy and multiple transfusions following trauma]. PMID- 11888504 TI - [Osteoporosis: risk factors and bone densitometry]. PMID- 11888506 TI - Dynamics of reverse cholesterol transport: protection against atherosclerosis. AB - This review considers the antiatherogenic function of high density lipoprotein (HDL) from the point of view of its dynamics within the sequential steps of reverse cholesterol transport (RCT). It is postulated that the efficiency of cholesterol flux through the RCT pathways is clinically more relevant than the HDL cholesterol concentration. The particular role of pre-beta(1)-HDL is reviewed drawing attention to the relationship between its concentration and the flux of cholesterol through the RCT system. PMID- 11888507 TI - A novel chemokine, Leukotactin-1, induces chemotaxis, pro-atherogenic cytokines, and tissue factor expression in atherosclerosis. AB - Chemokines such as monocyte chemoattractant protein (MCP) -1 and interleukin (IL) 8 are known to be involved in various processes in atherosclerosis such as plaque formation, plaque rupture, and thrombus formation. We investigated whether a new chemokine, Leukotactin (LKN)-1, is involved in atherosclerosis. We tested the expression of LKN-1 by immunohistochemical methods in carotid atherosclerotic plaque specimen. Induction of pro-inflammatory cytokines, transmigration, and tissue factor (TF) expression were tested in THP-1 cells and human peripheral blood monocytes treated with recombinant human LKN-1. Immunohistochemical analyses revealed that expression of LKN-1 occurs in regions of plaques rich in foam cells. In a Boyden chamber assay, THP-1 cells treated with 0.01--10 nM of LKN-1 transmigrated through gelatin coated filters in a dose dependent manner. LKN-1 also induced the transient expression of TNF-alpha, IL-8, and MCP-1 within 15 min of the treatment of the THP-1 cells. When peripheral blood monocytes were treated with LKN-1, expression levels of TF and TF-mediated procoagulating activity were induced in a time- and dose-dependent manner. These results raise the possibility that LKN-1 is another chemokine that is involved in the atherogenesis. LKN-1 may chemoattract immune cells into the plaque, induce pro inflammatory cytokines, and produce thrombi by inducing TF expression. PMID- 11888508 TI - Elevated levels of plasma prekallikrein, high molecular weight kininogen and factor XI in coronary heart disease. AB - Increased levels of hemostatic factors may play a role in the pathogenesis of myocardial infarction by triggering thrombin formation. We measured factor XII (FXII), factor XI (FXI), plasma prekallikrein (PK) and high-molecular-weight kininogen (HK) in 200 patients having survived myocardial infarction for at least 2 months, and in 100 healthy controls. We found significantly elevated levels of FXI clotting activity (FXI:C), HK:C and of the amidolytic activity of PK (PK:Am) among the patients as compared to the controls. Plasma levels of FXI:C, HK:C and PK:Am in the highest quartile were associated with an odds ratio of 1.9 (95% CI: 1.0-3.8), 2.0 (95% CI: 1.0-4.0) and 5.4 (95% CI: 2.6-11.2), respectively, compared to the respective plasma levels in the lowest quartile. After correction for established clinical and laboratory risk factors, the association between PK:Am plasma levels and myocardial infarction remained significant (P=0.0007). Combination of high PK:Am plasma levels and smoking or arterial hypertension, respectively, resulted in a more than additive relative risk for myocardial infarction. PMID- 11888509 TI - A novel cholesteryl ester transfer protein promoter polymorphism (-971G/A) associated with plasma high-density lipoprotein cholesterol levels. Interaction with the TaqIB and -629C/A polymorphisms. AB - The plasma cholesteryl ester transfer protein (CETP) plays a key role in reverse cholesterol transport (RCT) by mediating the transfer of cholesteryl ester (CE) from high-density lipoprotein (HDL) to atherogenic ApoB-containing lipoproteins, including VLDL, IDL and LDL. We describe a new polymorphism located at position 971 in the human CETP gene promoter, which corresponds to a G/A substitution at a potential AvaI restriction site. The relationship between the -971G/A polymorphism, plasma lipid parameters and plasma CETP concentration was evaluated in the Etude Cas-Temoins de l'Infarctus du Myocarde (control-myocardial infarction cases) cohort, and revealed that the -971G/A polymorphism (A allele frequency: 0.491) was significantly associated with both plasma high-density lipoprotein cholesterol (HDL-C) levels and CETP concentration (P=0.006 and 0.009, respectively). Subjects with genotype -971GG displayed both low HDL-C levels and high plasma CETP concentration, while genotype -971AA subjects displayed the inverse relationship. Evaluation of potential interactions between the -971G/A and the -629C/A or TaqIB polymorphisms demonstrated that the -971G/A polymorphism interacts significantly with the functional -629C/A site and the TaqIB polymorphism with respect to plasma HDL-C levels (P=0.0014 and 0.012, respectively), but does not affect plasma CETP concentration. These results clearly suggest that the interaction between the 971G/A polymorphism and either the -629C/A or the TaqIB polymorphism on plasma CETP concentration is different than that implicated in HDL-C levels. Transient transfection of HepG2 cells revealed that the -971G/A polymorphism did not modulate transcriptional activity of the human CETP gene promoter. The -971G/A promoter polymorphism therefore constitutes a non-functional marker. Furthermore, the observed effects of the 971G/A polymorphism on both plasma CETP concentration and HDL-C levels are due to functional variants in linkage disequilibrium with it. Our findings strongly suggest the existence of as yet unidentified functional polymorphisms in the CETP gene promoter that could explain the association between specific polymorphisms of the CETP gene and both plasma HDL-C and CETP concentrations. PMID- 11888511 TI - Statins inhibit A beta-neurotoxicity in vitro and A beta-induced vasoconstriction and inflammation in rat aortae. AB - Freshly solubilized A beta peptides synergistically increase the magnitude of the constriction induced by endothelin-1 (ET-1), via the activation of a pro inflammatory pathway. We report that mevinolin and mevastatin, two inhibitors of the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase are able to completely abolish the vasoactive properties of A beta in rat aortae. Mevinolin also appears to oppose the increased vascular reactivity to ET-1 induced by interleukin 1-beta and phospholipase A(2) suggesting that statins display some anti-inflammatory properties. We show that freshly solubilized A beta stimulates prostaglandin E(2) and F(2 alpha) production (by 6 and 3.6 times, respectively) in isolated rat aortae and that mevinolin completely antagonizes this effect confirming the anti-inflammatory action of mevinolin ex vivo in rat aortae. In addition, we observed that A beta vasoactivity is not mediated nor modulated by mevalonic acid suggesting that the anti-inflammatory action of the statins are not related to an inhibition of HMG-CoA reductase activity. Differentiated human neuroblastoma cells (IMR32) were used to assess the neurotoxic effect of pre aggregated A beta by quantifying the release of lactate dehydrogenase (LDH) in the cell culture medium. A beta appears to enhance LDH release by 30% in IMR32 cells, an effect that can be completely opposed by mevastatin. Taken together these data show that statins can antagonize the effect of A beta in different assays and provide new clues to understand the prophylactic action of the statins against Alzheimer's disease. PMID- 11888510 TI - Expression of transcriptional repressor ATF3/LRF1 in human atherosclerosis: colocalization and possible involvement in cell death of vascular endothelial cells. AB - Vascular endothelial cell death contributes to the progression of atherosclerotic lesion, and several transcriptional regulators are involved in the process. Activating transcription factor 3/liver regenerating factor-1 (ATF3/LRF-1), a stress-inducible transcriptional repressor, was shown to be highly expressed in vascular endothelial cells and macrophages of human atherosclerotic lesions by immunohistological assay. The expression was colocalized in these cells which were positive for TdT-mediated dUTP nick-end labeling (TUNEL) and annexin V. Treatment of human umbilical vein endothelial cells (HUVECs) by tumor necrosis factor (TNF)-alpha, oxidized low density lipoprotein (oxLDL), and lysophosphatidylcholine (LPC) rapidly induced ATF3/LRF-1, which showed an increased DNA binding to the consensus ATF/CRE sequence by supershift of gel shift assay. Flow cytometry analysis and immunostaining analysis with TUNEL assay showed that ATF3/LRF-1 was highly expressed in cell death induced by these agents. Moreover, antisense ATF3/LRF-1 cDNA partly suppressed the cell death induced by TNF-alpha, oxLDL, and LPC. From these results, it is indicated that ATF3/LRF-1 is one of the immediate early response genes in vascular endothelial cells in response to atherogenic stimuli, and may play a role in the endothelial cell death associated with atherogenesis. PMID- 11888512 TI - The effects of hydroxy-methyl-glutaryl co-enzyme A reductase inhibitors on platelet thrombus formation. AB - BACKGROUND: Hydroxy-methyl-glutaryl co-enzyme A reductase inhibitors (HMG CoA RIs) markedly improve the lipid profile of patients with hypercholesterolemia, but the magnitude and time course of the effect of these drugs on other risk factors for atherosclerosis are not well defined. METHODS: We employed a random assignment, double-blind design to compare the effect of 8 weeks of HMG CoA RI therapy with either pravastatin (40 mg QD; n=12) or simvastatin (20 mg QD; n=12) with placebo (n=13) on serum lipids, platelet thrombus formation (PTF), and markers of inflammation and thrombosis in patients with coronary artery disease. PTF was measured using a validated ex vivo perfusion chamber system. RESULTS: Total and LDL cholesterol decreased 20.3 +/- 12.7% and 31.4 +/- 16.5% in the HMG CoA RI group and were unchanged with placebo (P<0.01). Triglycerides also decreased 15.3 +/- 22.5% with HMG CoA RI therapy, but increased 8.4 +/- 30.0% with placebo (P=0.01). PTF increased 54.1 +/- 89.0% with placebo and decreased 8.0 +/- 46.82% with HMG CoA RI treatment (P<0.01). CONCLUSIONS: HMG CoA RI therapy with pravastatin or simvastatin reduces PTF after only 8 weeks of therapy. Such lipid effects may contribute to the prompt reduction in cardiovascular events noted in some clinical trials. PMID- 11888513 TI - Oxidative stress increases the expression of the CD36 scavenger receptor and the cellular uptake of oxidized low-density lipoprotein in macrophages from atherosclerotic mice: protective role of antioxidants and of paraoxonase. AB - Little is known about the effects of oxidative stress on macrophage lipid peroxidation and on their atherogenic consequences. Therefore, we questioned the causal relationship between cellular lipid peroxides content and macrophage uptake of oxidized low-density lipoprotein (Ox-LDL). Lipid peroxide content in mouse peritoneal macrophages (MPMs) from E-deficient (E(0)) mice increased progressively by up to 4.6 fold during mice aging, and this was accompanied by an age-dependent increase in the cellular uptake of Ox-LDL (90%), and in the expression of the scavenger receptor CD36 mRNA (41%). Inhibition or stimulation of cellular oxidative stress by administration of dietary potent antioxidants (vitamin E or glabridin) or by inducing cellular glutathione depletion (by using buthionine sulfoximine), respectively, resulted in a significant increment or inhibition of macrophage uptake of Ox-LDL and in cellular CD36 mRNA expression, respectively. Intraperitoneal injection of human serum paraoxonase (PON1) into E(0) mice, resulted in a 40-65% decrement in the lipid peroxide content in MPM harvested from E(0) mice at 2-5 months of age, which subsequently resulted in a similar reduced uptake of Ox-LDL and expression of CD36 mRNA (by 30-40%). In conclusion, our results are the first to demonstrate that macrophage lipid peroxidation stimulates CD36 mRNA expression and enhances the cellular uptake of Ox-LDL. PMID- 11888514 TI - The TNF alpha/G-308A polymorphism influences insulin sensitivity in offspring of patients with coronary heart disease: the European Atherosclerosis Research Study II. AB - There is accumulating evidence for a role of tumor necrosis factor-alpha (TNF alpha) in insulin resistance induced by obesity. The purpose of this study was to investigate whether the TNF alpha/G-308A polymorphism was associated with responses to oral glucose and fat tolerance tests in a case--control study comparing male offspring with a paternal history of premature myocardial infarction (cases, n=335) to age-matched controls (n=340) recruited from 14 European university populations. Genotype frequencies did not significantly differ between cases and controls. Among cases, those carrying the A allele exhibited a higher area under the curve for insulin (64.5 vs 55.9 mU h/l, P=0.009), a higher increment between baseline concentration and peak of insulin (63.1 vs 52.8 mU/l, P=0.005) and a greater decrease between peak and insulin at 120 min (49.1 vs 36.8 mU/l, P=0.003) than those with the GG genotype. No such effect was observed in control subjects. No association was observed with response to a fat tolerance test either in cases or in controls. The present results suggest that the TNF alpha/G-308A polymorphism might interact with other susceptibility factors to coronary heart disease to predispose to insulin resistance, and that the ability of TNF-alpha to induce insulin resistance may extend beyond obesity. PMID- 11888516 TI - Genetic determination of HDL variation and response to diet in baboons. AB - We fed 634 baboons three diets to assess the separate effects of increasing dietary fat and cholesterol intakes on three independent measures of HDL phenotype: concentrations of HDL cholesterol and apoAI, and size distributions of HDL cholesterol. Increasing dietary fat significantly increased concentrations of HDL cholesterol and apoAI (both, P<0.0001), but did not affect HDL particle sizes, whereas increasing dietary cholesterol increased HDL cholesterol (P<0.0001) concentrations and HDL particle sizes (P=0.08), but did not affect apoAI concentrations. A substantial proportion of variation in each of the HDL traits was influenced by genes (heritabilities ranged from 25 to 61%) and a common set of genes influenced HDL variation on each of the diets (genetic correlations ranged from 0.64 to 1.0). However, genes exerted a smaller effect on HDL response to changes of dietary fat and of dietary cholesterol. Therefore, dietary fat and cholesterol alter HDL levels and characteristics, but the dietary responses are not strongly mediated by additive genetic effects. PMID- 11888515 TI - Dietary antioxidants preserve endothelium dependent vasorelaxation in overfed rats. AB - OBJECTIVE: A high fat diet contributes to obesity and acutely impairs endothelium dependent vasorelaxation. While a high cholesterol diet chronically impairs endothelium dependent vasorelaxation in rabbits, this model is associated with severe hypercholesterolemia. The effect of chronic high fat feeding on endothelial function in the setting of more normal lipid levels has not been studied. Our aim was to study vascular function in rats overfed for 6 months and to determine the role of oxidative stress in the alteration of vascular function associated with this diet. METHODS: Forty-five male Sprague-Dawley rats were placed on the following diets for 6 months, control diet, high fat diet or high fat diet supplemented with vitamins A, E and selenium. Six months later blood samples were collected and vascular function was assessed in the aorta. RESULTS: The rats fed a high fat diet were heavier than controls (608.4 +/- 41.8 vs. 700.3 +/- 50.1 vs. 699.5 +/- 52.6 g, P<0.05 for control vs. high fat and high fat plus antioxidant groups) but lipid levels were similar in each group (cholesterol, 145.9 +/- 53.4 vs. 140.5 +/- 44.0 vs. 152.7 +/- 36.1 mg/dl and triglycerides, 173.2 +/- 106.7 vs. 197.4 +/- 131.3 vs. 166.1 +/- 65.3 mg/dl, P, NS). Relaxations to acetylcholine and calcium were significantly impaired in the high fat diet group compared with controls (EC(50), 6.90 +/- 0.22, 7.12 +/- 0.32; AUC, 96.9 +/- 51.6, 155.5 +/ - 73.7) but were not different between the antioxidant supplemented group and controls (EC(50), 7.06 +/- 0.37; AUC, 151.9 +/- 67.4). Relaxations to DEA NONOate were similar in each group. CONCLUSIONS: Dietary antioxidants preserved endothelium dependent vasorelaxation in rats fed a high fat diet for 6 months. PMID- 11888517 TI - The effect of chronic insulin delivery via the intraperitoneal versus the subcutaneous route on hepatic triglyceride secretion rate in streptozotocin diabetic rats. AB - Chronic intraperitoneal or subcutaneous insulin administration increases triglyceride secretion rate (TGSR) in normal rats. We wished to determine the effect of this treatment on TGSR and the hepatic lipogenic enzymes acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS) in diabetic rats. Streptozotocin diabetic rats, untreated (D), diabetic rats treated with insulin (3 U/day for 21 days) intraperitoneally (IP) or subcutaneously (SC) and non-diabetic rats (N) were studied. TGSR was determined using Triton WR-1339. Fasting glucose and triglyceride levels, high in D, were normalized by insulin treatment regardless of route. Peripheral insulin levels were lowest in D and highest in SC, portal insulin levels were lowest in D and highest in IP. Non-esterified fatty acid levels were not elevated in D, presumably due to adipose tissue depletion. TGSR was reduced in D (P<0.05) and was normalized following insulin administration, regardless of route. ACC activity was normal, but FAS was decreased in D (P<0.05). ACC and FAS were normal in both IP and SC. Thus, in streptozotocin diabetic rats, chronic intraperitoneal or subcutaneous insulin treatment increases TGSR and FAS activity from their low levels in insulin-deficient rats to levels equal to but not higher than those in normal rats. PMID- 11888518 TI - Effect of BO-653 and probucol on c-MYC and PDGF-A messenger RNA of the iliac artery after balloon denudation in cholesterol-fed rabbits. AB - Antioxidants have been proposed as a promising treatment for restenosis after percutaneous transluminal coronary angioplasty (PTCA), but their mechanism of action remains unclear. Here, we investigated the effect of antioxidants on gene expression in the artery after balloon denudation. We developed a sensitive ribonuclease (RNase) protection assay for the messenger RNA (mRNA) levels of immediate early (IE) genes (c-jun, c-fos and c-myc), as well as platelet-derived growth factor-A (PDGF-A), platelet-derived growth factor-beta receptor, transforming growth factor-beta 1, and vascular endothelial growth factor. New Zealand White rabbits were fed a 0.17% cholesterol diet containing vehicle, BO 653 or probucol, and balloon denudation for iliac arteries was performed. The iliac arteries were then removed at 4 h after the denudation, for IE genes, and 10 days after for growth factors and receptors. Both BO-653 and probucol significantly reduced neointimal thickening, compared with the control. In terms of gene expression, BO-653, but not probucol, significantly inhibited c-myc induction. On the other hand, probucol, but not BO-653, significantly inhibited PDGF-A expression. Neither treatment had any effect on the expression of other genes. These results suggest that antioxidants affect the gene expression of the neointimal response and that both BO-653 and probucol inhibit gene expression in specific manners. PMID- 11888520 TI - Endothelial dysfunction in hypercholesterolemia is improved by L-arginine administration: possible role of oxidative stress. AB - Hypercholesterolemia impairs endothelial function. However, the production/release of nitric oxide from the hypercholesterolemic aorta is reported to be enhanced rather than impaired in animal studies. L-arginine improves endothelial function in hypercholesterolemic subjects. The goal of the present study was to investigate the effects of L-arginine on endothelial function and oxidative stress in hypercholesterolemic subjects. In 17 hypercholesterolemic male subjects (mean age 41.7 years, mean total cholesterol 264.3 +/- 5.9 mg/dl) and 17 age-matched healthy men as controls (mean total cholesterol 187.1 +/- 6.8 mg/dl), we measured flow-mediated endothelium-dependent vasodilation of the brachial artery during saline infusion and after saline plus L-arginine infusion (30 g for 1 h) with ultrasound technique. In addition, we measured the levels of thiobarbituric acid reactive substances (TBARS), as a marker of lipid peroxide. The flow-mediated vasodilation was lower and the TBARS concentration was higher in the hypercholesterolemic group than in the control group during the saline infusion. The addition of L-arginine increased flow mediated vasodilation and decreased TBARS concentration in the hypercholesterolemic group (from 3.92 +/- 0.58 to 7.27 +/- 0.53% [P<0.01 by analysis of variance (ANOVA)], from 7.74 +/- 0.46 to 5.71 +/- 0.35 nmol/ml [P<0.01 by ANOVA], respectively), but not in the control group (from 7.74 +/- 0.40 to 8.21 +/- 0.47%, from 5.45 +/- 0.43 to 4.83 +/- 0.35 nmol/ml, respectively). The endothelial function is blunted, and the oxidative stress is increased in hypercholesterolemic subjects. L-arginine improves endothelial function with decreasing oxidative stress. The augmentation of nitric oxide production/release induced by L-arginine may act as an antioxidant, and contributes to the improvement of endothelial function in hypercholesterolemic subjects. PMID- 11888519 TI - Homocysteine increases monocyte and T-cell adhesion to human aortic endothelial cells. AB - Although hyperhomocysteinemia has been recognized as an independent risk factor for atherosclerosis, its mechanism(s) are not well understood. Because chemotaxis and accumulation of leukocytes such as monocytes and T cells have been demonstrated to be critical events in the initiation and development of atherosclerosis, we investigated the effect of homocysteine (HCY) on U937 monocytic cells- and Jurkat T-cell-human aortic endothelial cell (HAEC) interactions under inflammatory cytokine-stimulated conditions. When HAEC were pretreated with HCY followed by stimulation with IL-1 beta, U937 and Jurkat T cell adhesion to HAEC increased in a dose-dependent manner. The significant increase in U937 cell adhesion to HAEC was also observed when U937 cells were treated with HCY or when both cell types were treated with HCY. We also demonstrated that HCY increases endothelial surface expression and mRNA level of adhesion molecules, VCAM-1 and E-selectin. Attenuation of Jurkat T-cell and U937 cell adhesion to HAEC by monoclonal antibodies directed to specific adhesion molecules demonstrated that both VCAM-1 and E-selectin are involved in Jurkat T cell adhesion, and VCAM-1 in U937 cell adhesion. Supplementation of HAEC with vitamin E was effective in preventing HCY-stimulated Jurkat T-cell adhesion and VCAM-1 and E-selectin expression in HAEC. These results indicate that HCY mediated leukocyte-endothelial cell interaction is one potential mechanism by which homocysteinemia may lead to the development of atherosclerosis under inflammatory conditions. Dietary antioxidants such as vitamin E may attenuate HCY stimulated activation of the endothelium and may help reduce the risk of vascular disease associated with hyperhomocysteinemia. PMID- 11888521 TI - Inhibition of TNF-alpha induced ICAM-1, VCAM-1 and E-selectin expression by selenium. AB - The initiation of an atherosclerotic lesion involves an endothelial cell pro inflammatory state that recruits leukocytes and promotes their movement across the endothelium. These processes require endothelial expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial-leukocyte adhesion molecule-1 (E-selectin). Tumor necrosis factor alpha (TNF-alpha) is a powerful inducer of these adhesion molecules. Selenium status is known to affect the rate of atherosclerosis. These experiments tested whether selenium alters cytokine-induced expression of these adhesion molecules. Human umbilical vein endothelial cells (HUVECs) were pretreated for 24 h with sodium selenite (0-2 microM) and then treated with 0 or 50 U/ml TNF-alpha in the presence of 0-2 microM selenite. ICAM-1, VCAM-1 and E-selectin were detected by ELISA and their mRNAs were evaluated by Northern blots. Selenite significantly inhibited TNF-alpha-induced expression of each adhesion molecule in a dose dependent manner and reduced the level of the respective mRNAs. Nuclear factor kappa B (NF-kappa B) is required for transcription of these adhesion molecule genes. Western blot analysis revealed that selenite did not inhibit the translocation of the p65 subunit of NF-kappa B to the nucleus. In conclusion, these data indicate selenium can modulate cytokine-induced expression of ICAM-1, VCAM-1 and E-selectin in HUVECs without interfering with translocation of NF kappa B. PMID- 11888522 TI - Lysophosphatidylcholine induces apoptosis in human endothelial cells through a p38-mitogen-activated protein kinase-dependent mechanism. AB - Lysophosphatidylcholine (lysoPC) is a component of oxidized low density lipoprotein (LDL) and is involved in the pathogenesis of atherosclerosis and inflammation. Previous studies demonstrated that lysoPC can induce various protein kinases including tyrosine kinases, protein kinase C (PKC), and mitogen activated protein kinases (MAPK) in vascular endothelial cells. However, the role of lysoPC-activated kinases remains undefined. In this study, we examined the effect of lysoPC on apoptosis and investigated the role of lysoPC-activated protein kinases in human umbilical vein endothelial cells (HUVEC). The presence of apoptosis was evaluated by morphological criteria, MTT assay, and electrophoresis of DNA fragments showing the characteristic apoptotic ladder, TUNEL analysis, and quantified as the proportion of hypodiploid cells by flow cytometry. The lysoPC induced apoptosis in a time- and dose-dependent manner. It stimulated the phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2) and p38-MAPK in HUVEC. The use of specific pharmacologic inhibitors indicated that the p38-MAPK-signaling pathway (SB203580) is required for lysoPC induced apoptotic signals. Furthermore, lysoPC-induced apoptosis was inhibited by DEVD-FMK (a caspas-3/CPP32 inhibitor), suggesting involvement of an important segment in the apoptosis. These results demonstrate that lysoPC induces apoptosis in human endothelial cells through a p38-MAPK-dependent pathway. PMID- 11888523 TI - Beneficial effects of blood donation on high density lipoprotein concentration and the oxidative potential of low density lipoprotein. AB - According to some authors blood donors have a lower risk of cardiovascular incidents. This may be associated with the risk of cardiovascular disease reported by some authors, as well as with the oxidative changes caused by iron. The aim of this study was to determine, what happens to some of the factors contributing to atherosclerosis after the lowering of body iron. Blood was drawn from 23 healthy males after overnight fasting and the parameters described below determined. These persons donated blood (500 ml) on three occasions with 6 weeks intervals. Six to eight weeks after the third and final donation, blood was again drawn after overnight fasting and the following parameters measured for the second time: various parameters of body iron; lipid profile; anti-oxidants; and oxidative parameters of low density lipoprotein (LDL). Blood donation has various beneficial effects, such as increasing high density lipoprotein (HDL) and apoA; a higher oxidative potential of LDL; a lower level of LDL peroxidation resulting in a LDL particle with a higher oxidative potential, and a higher NO(3) concentration. We conclude that blood donation, and thereby a lowered body iron concentration, is an effective way to increase the oxidative potential of LDL, as well as the HDL and apoA concentrations. PMID- 11888524 TI - Increased levels of monocyte-related cytokines in patients with unstable angina. AB - Inflammatory cytokines play important roles in coronary artery disease. We investigated the clinical significance of monocyte-related cytokine expression in patients with angina pectoris. We studied 26 patients with stable effort angina and 20 patients with unstable angina in whom stenotic lesions of the coronary arteries were confirmed by selective coronary angiography. Plasma levels of interleukin-6 (IL-6), macrophage colony stimulating factor (MCSF), and monocyte chemoattractant protein-1 (MCP-1) were measured. Plasma levels of IL-6, MCSF, and MCP-1 in patients with unstable angina were significantly higher than those in patients with stable angina or control subjects. Patients with unstable angina were further divided into sub-groups according to their clinical classification; Levels of IL-6, MCSF, and MCP-1 in patients, who had anginal attacks at rest within the 48 h prior to admission (Braunwald class IIIB) were significantly higher than those in patients, who did not have attacks at rest (class IB). Five unstable patients, who were refractory to medical therapy and were referred for emergency coronary revascularization showed marked elevation of plasma MCSF and MCP-1 levels. In conclusion, plasma levels of monocyte-related cytokines were elevated in unstable angina. These increases were marked in patients with unstable angina with recent ischemic attack at rest, suggesting that activation of monocytes is involved in vulnerability of underlying atheromatous plaque. PMID- 11888525 TI - Effects of menopause on intraindividual changes in serum lipids, blood pressure, and body weight--the Chin-Shan Community Cardiovascular Cohort study. AB - In Taiwan, the Chin-Shan Community Cardiovascular Cohort (CCCC) was assessed prospectively to determine whether the changes in cardiovascular risk factors for women age 45--54 years are due to menopause. The average paired percentage changes that occurred between baseline (1990-1991) and follow-up (4 years later) in fasting serum lipids were compared in three groups of women including groups of 59 and 224 who were pre- and postmenopausal, respectively, and a group of 118 who had spontaneously stopped menstruating. Postmenopausal women had the least gain in body mass index (BMI), whereas, mainly premenopausal women had increased systolic blood pressure (P<0.05). All women had elevated total cholesterol (TC) levels, with the greatest elevation in women transitioning into menopause (P<0.001). Low-density lipoprotein cholesterol (LDL-C) levels increased before and during the transition to menopause, but decreased after menopause (P<0.01). Age had significant association with changes in TC, triglyceride (TG) and LDL-C levels, whereas BMI had significant association with changes in TG, LDL-C, and high-density lipoprotein cholesterol (HDL-C) levels (P<0.05). After controlling for age and BMI, only differences in TC remained significant, with the greatest gain in women who stopped menstruating (12.9%) followed by pre- (6.5%) and postmenopausal women (4.8%). Changes in both systolic and diastolic blood pressures, and TG and HDL-C levels were not significantly different, but HDL-C levels declined between 11.5 and 14.7% in all groups. This study suggests an unfavorable effect of menopause on lipid metabolism, especially on the TC level, which was predominantly elevated during the transition to menopause. The decline of HDL-C is of concern. PMID- 11888526 TI - Polymorphism of HL +1075C, but not -480T, is associated with plasma high density lipoprotein cholesterol and apolipoprotein AI in men of a Chinese population. AB - Eight hundred and twenty-three Chinese Han adults aged over 40, including 466 men and 357 women were enrolled in the study to examine the association of +1075C and -480T polymorphisms in hepatic lipase gene with plasma lipoprotein and apolipoprotein levels. In this population the allele frequencies for HL +1075C and minus -480T were 0.053 and 0.362, respectively and the prevalence of HL +1075C/C and -480T/T were 0.006 and 0.132, respectively. Overall, the normal HDL C (> or = 35 mg/dl) subjects had a higher carrier frequency of HL +1075C than the low HDL-C (<35 mg/dl) subjects (0.108 vs 0.029, P=0.039). However, when tested separately, the carrier frequency of HL +1075C was not significantly different between normal and low HDL-C females (0.101 vs 0.083, P=0.843). In males, the normal HDL-C subjects had a higher carrier frequency of HL +1075C than the low HDL-C subjects (0.113 vs 0.018, P=0.026). No significant difference of frequencies of HL -480T genotypes -480T/T,-480C/T and -480C/C was found between normal and low HDL-C subjects. Among plasma TG, TC, HDL-C, apo AI, apo AII, apo B100, apo CII, Apo CIII and apoE, only HDL-C and apo AI were significantly different among the three genotypes +1075A/A,+1075A/C and +1075C/C in men (P=0.029 and 0.032). No association was found in women. Male subjects with CC had a higher HDL-C than those with AC (P=0.020) and AA (P=0.013), AC higher than AA (P=0.017). Male subjects with CC had a higher apo AI than AC (P=0.013) and AA (P=0.019), AC higher than AA (P=0.021). Although not so significant (P=0.053) as HDL-C and apo AI, male subjects with CC had a higher apo AII than those with AC and AA. No significant difference of lipoprotein and apolipoprotein traits was found among the three -480T genotypes -480C/C,-480C/T and -480T/T, in the sample overall and in men and women separately. These results indicate that HL +1075C, not -480T polymorphism is associated with plasma high density lipoprotein cholesterol and apolipoprotein AI in men in this Chinese population. PMID- 11888527 TI - Differential effect of two common polymorphisms in the cholesteryl ester transfer protein gene on low-density lipoprotein particle size. AB - This study was performed to determine the relationship between the two common polymorphisms of cholesteryl ester transfer protein (CETP) gene and LDL size in Japanese individuals. The LDL size was determined by gradient gel electrophoresis in 136 patients undergoing routine check-ups. The presence of two polymorphisms (I405V and Taq1B) was determined using PCR-based methods. The VV genotype for the I405V polymorphism was associated with both a lower plasma CETP concentration and a higher plasma HDL-C concentration. Further, the LDL size in patients with the VV genotype was significantly smaller than that in patients with the II+IV genotype (26.0 +/- 0.8 vs. 26.3 +/- 0.7 nm, P<0.05). Although the B2B2 genotype for the Taq1B polymorphism was also associated with both a lower plasma CETP concentration and a higher plasma HDL-C concentration, it had no effect on the LDL size (26.2 +/- 0.7 vs. 26.3 +/- 0.8 nm, P=0.73). Stepwise multiple regression analysis revealed that the VV genotype, as well as plasma TG concentration, age, HbA1c concentration, and BMI, were determinants of LDL size, while no significant relationships were seen between any of the Taq1B polymorphisms and LDL size. These data suggest that the I405V polymorphism but not the Taq1B polymorphism may be responsible for the distribution of LDL size. This may explain the differential effects of these two polymorphisms on the risk of CHD. PMID- 11888528 TI - Hypertriglyceridemia is associated with an elevated blood viscosity Rosenson: triglycerides and blood viscosity. AB - Elevated blood viscosity is a predictor of cardiovascular disease. The major determinants of blood viscosity are hematocrit and plasma viscosity. Plasma triglycerides elevate plasma viscosity; however, the contribution of plasma triglycerides to blood viscosity after adjustment for other major covariates has not been reported. This cross-sectional study of 257 adult subjects evaluated the associations between fasting plasma lipids, fibrinogen, total serum protein, hematocrit and blood viscosity. Blood viscosity was measured at 37 degrees C with a coaxial cylinder microviscometer at shear rates of 100 and 1 s(-1). Blood viscosity values are reported both as uncorrected measurements and measurements corrected to a hematocrit of 45% by a regression equation. Uncorrected blood viscosity at a shear rate of 100 s(-1) was significantly associated with triglycerides, fibrinogen, high density lipoprotein (HDL) cholesterol, total serum protein, and hematocrit using stepwise multivariate regression analysis. When corrected blood viscosity at 100 s(-1) was the dependent variable, there were statistically significant associations with triglycerides, HDL cholesterol, and total serum protein. Corrected blood viscosity at 1 s(-1) was significantly associated with triglycerides, fibrinogen, total serum protein, and an indicator variable for diabetes mellitus. This study supports an additional mechanism whereby triglycerides may contribute to cardiovascular risk. PMID- 11888529 TI - Haptoglobin phenotype and coronary artery collaterals in diabetic patients. AB - Cross-cultural epidemiological studies of incident cardiovascular disease in the diabetic patient have demonstrated marked differences in susceptibility that may be due to a genetic factor. The coronary artery collateral circulation is the chief determinant of the size of a myocardial infarction and is highly variable between patients. We recently demonstrated that a functional allelic polymorphism in the haptoglobin gene is correlated with a number of diabetic vascular complications. We thus set out to test the hypothesis that haptoglobin phenotype is associated with collateral formation in the setting of diabetes. We correlated the Hp phenotype (1-1, 2-1 or 2-2) as determined by polyacrylamide electrophoresis with the presence or absence of coronary collaterals by angiography in 82 consecutive diabetic patients and 138 consecutive non-diabetic patients undergoing catheterization. We found that diabetic patients with the Hp phenotype 2-1 were more likely to have collaterals than diabetic patients with the Hp phenotype 2-2 (P=0.007). There was no correlation between Hp phenotypes and the presence of collaterals in non-diabetic patients. Hp phenotype thus appears to be associated with the development of the coronary collateral circulation in diabetic patients with coronary artery disease. Haptoglobin 2-2 may predispose to less compensation for coronary artery stenosis in diabetic patients, and thereby portend a worse prognosis. PMID- 11888530 TI - LDL concentration is correlated with the removal from the plasma of a chylomicron like emulsion in subjects with coronary artery disease. AB - In animal model studies, the uptake of chylomicron remnants after entering in the space of Disse occurs mainly by low-density lipoprotein (LDL) receptor and LDL receptor-related protein (LRP). In subjects, the relative importance of each one of these receptors for the clearance of chylomicron remnants is not fully understood. In our study, LDL cholesterol and apolipoprotein (apo) B were correlated to the plasma kinetics of a chylomicron-like emulsion in 77 subjects (11 women, mean age 58 +/- 12 years) with coronary artery disease (CAD). Their total cholesterol was 227 +/- 25 mg/dl, triglyceride 159 +/- 25 mg/dl, LDL cholesterol 148 +/- 27 mg/dl, HDL cholesterol 40 +/- 9 mg/dl, apo A1 1.80 +/- 0.53 g/l and apo B 1.65 +/- 0.48 g/l. The emulsion was double-labeled with 3H triolein and 14C-cholesteryl oleate and injected intravenously after 12-h fasting. The decay curves of the radioisotopes were determined from blood samples collected at predetermined intervals during 60 min. A negative correlation between FCR of the emulsion cholesterol esters and LDL cholesterol and apo B plasma concentrations was found (r=-0.4, P=0.005 and r=-0.3, P=0.01, respectively) whereas FCR of the emulsion triglycerides did not correlate with any of the plasma lipids or apolipoprotein parameters. Concluding, in patients with CAD, LDL catabolic pathway significantly influences the removal from plasma of chylomicron remnants. PMID- 11888531 TI - Sex hormone-binding globulin levels and cardiovascular risk factors in morbidly obese subjects before and after weight reduction induced by diet or malabsorptive surgery. AB - One of the main goals of weight reduction in morbidly obese subjects is its benefit on coronary heart disease (CHD) risk. A cross-sectional study was designed to randomly assign 79 morbidly obese subjects (27 men and 52 women; age: 30-45 years) either to a diet protocol (20 kcal per kg fat-free mass (FFM); 55% carbohydrates, 30% fat, and 15% proteins) or to malabsorptive surgery (biliopancreatic diversion). Fatness parameters, measured by dual-energy X-ray absorptiometry, lipid profile, insulin, leptin, sex steroid hormones and sex hormone-binding globulin (SHBG) levels were compared at baseline and 1 year after the beginning of the study. The data showed that plasma SHBG levels, but not testosterone levels, correlated negatively to fasting insulin levels and positively to HDL-cholesterol in both men and women. Total leptin levels were significantly lower (P<0.0001) in post-BPD subjects of both sexes compared to dietary treated obese subjects. The logarithm of plasma leptin correlated significantly and positively with insulin but negatively with SHBG.A step-down regression analysis showed that FFM and SHBG, but not insulin levels, were the most powerful independent variables for predicting HDL-cholesterol levels in morbidly obese patients. The negative relationship between SHBG levels and CHD risk appears to be mediated by a concomitant variation in body fatness. Finally, in obese patients, SHBG levels seem to be an indicator of total adiposity rather than an index of an altered insulin/glucose homeostasis. PMID- 11888532 TI - High expressor paraoxonase PON1 gene promoter polymorphisms are associated with reduced risk of vascular disease in younger coronary patients. AB - Human paraoxonase-1 is hypothesised to protect serum lipoproteins from oxidative stress. Decreased serum activity of paraoxonase-1 in animal models is associated with an increased risk of vascular disease and has been linked to the anti oxidant capacity of the enzyme. Promoter polymorphisms of the human paraoxonase-1 gene strongly influence serum concentrations of the enzyme. The present study examined the hypothesis that promoter polymorphisms may be genetic risk factors for vascular disease in man. Genotypes arising from the promoter C(-907)G polymorphism were analysed in the ECTIM2 population. The global odds ratio for myocardial infarction, comparing the high expressor GG genotype to other genotypes, was 0.77 (0.61-0.97) (P=0.024). The association with the promoter genotype was more pronounced in the youngest age group (odds ratio 0.52 (0.31 0.87), P=0.012) and was progressively lost with age (respectively 50 years to <60 years, P=0.26; >60 years, P=0.45). There was no association between the promoter genotypes and serum lipids. The data are consistent with the high expressor promoter genotype being linked to reduced risk of myocardial infarction. The influence of the genotype may be compromised in older patients. PMID- 11888533 TI - A prospective study of TaqIB polymorphism in the gene coding for cholesteryl ester transfer protein and risk of myocardial infarction in middle-aged men. AB - BACKGROUND: Molecular variations in the gene coding for the cholesteryl ester transfer protein (CETP) such as the TaqIB polymorphism are associated with higher plasma high-density lipoprotein (HDL) concentration. However, whether this polymorphism is associated with risk of myocardial infarction (MI) is uncertain. METHODS AND RESULTS: In a prospective cohort of 14916 apparently healthy men enrolled in the Physicians' Health Study, allelic status for the TaqIB polymorphism in the CETP gene was determined among 384 participants who subsequently developed a first MI (cases) and among an equal number of age and smoking-matched participants who remained free of cardiovascular disease during follow-up (controls). Overall, the B2B2 genotype was present in 17% of the study participants and was associated with higher HDL cholesterol levels (mean mg/dl [+/- S.D.], 45 +/- 11 for the B1B1 genotype, 48 +/- 13 for the B1B2 genotype and 50 +/- 12 for the B2B2 genotype; P=0.01). However, the risk of developing MI did not differ significantly across these three genotypes. After adjustment for coronary risk factors (but not HDL), the relative risks for future MI were 1.12(95% CI 0.74-1.70) for the B1B2 genotype and 0.95(95% CI 0.54-1.66) for the B2B2 genotype, compared with the B1B1 genotype. In subgroup analysis of individuals with low HDL levels, B2B2 genotype appeared to have a lower risk of MI compared with the B1B1 genotype. However, participants with high HDL were at lower risk of developing MI regardless of their CETP genotype. CONCLUSIONS: In this prospective study of apparently healthy middle-aged US men, carriers of the B2 allele of the TaqIB in the CETP gene had higher HDL concentrations, but did not have lower risk of MI. CONDENSED ABSTRACT: In a cohort of apparently healthy middle-aged US men, the relation between CETP genotype and MI risk was prospectively examined in a nested case-control study. After adjusting for coronary risk factors (but not HDL), the 9-year risk of developing MI did not differ significantly by genotype. Comparing to the B1B1 genotype, the relative risks for future MI were 1.12 (95% CI 0.74-1.70) for the B1B2 genotype and 0.95 (95% CI 0.54-1.66) for the B2B2 genotype. PMID- 11888534 TI - Orientation sensitivity of ganglion cells in primate retina. AB - The two-dimensional shape of the receptive field center of macaque retinal ganglion cells was determined by measuring responses to drifting sinusoidal gratings of different spatial frequency and orientation. The responses of most cells to high spatial frequencies depended on grating orientation, indicating that their centers were not circularly symmetric. In general, center shape was well described by an ellipse. The major axis of the ellipse tended to point towards the fovea or perpendicular to this. Parvocellular pathway cells had greater center ellipticity than magnocellular pathway cells; the median ratio of the major-to-minor axis was 1.72 and 1.38, respectively. Parvocellular pathway cells also had centers that were often bimodal in shape, suggesting that they received patchy cone/bipolar cell input. We conclude that most ganglion cells in primate retina have elongated receptive field centers and thus show orientation sensitivity. PMID- 11888535 TI - Age-related changes in the visual cortex. AB - The ability to accurately perceive the speed of moving objects is one of many visual functions that decline with age. One factor that may contribute to this is a deterioration in temporal processing speed. At present, there is a dearth of information concerning how this may occur in the central nervous system, particularly in the visual cortex. Thus, in the present study, we investigated the neural basis of speed and temporal processing in areas 17 and 18 of visual cortex in young and aged rats using either a moving bar of light or a series of flashing lights. Our results showed that the mean preferred speed of a moving bar of light was significantly reduced in aged as compared to young animals. We also found that cells recorded from young animals were able to entrain to a higher frequency of flashing light stimuli than those recorded from aged animals. In addition, we found no age-related differences between cortical fields. These results suggest an age-related difference in temporal processing speed at the level of visual cortex. PMID- 11888536 TI - Orientation- and frequency-modulated textures at low depths of modulation are processed by off-orientation and off-frequency texture mechanisms. AB - Intuitively it may seem likely that orientation-modulated (OM) and frequency modulated (FM) textures are processed utilizing the first-order channels that are most responsive to the first-order (luminance) information contained in the textures. This assumption would imply that the detection or segmentation of OM or FM textures is accomplished by second-order mechanisms that receive their first order input from neurons tuned to either the center, or to the peaks in the orientation and spatial-frequency distribution of the texture. Here we show that at low depths of modulation this is not the case. Using an adaptation paradigm, we show that the first-order filters involved in the perception of OM and FM textures are those which maximize the differential response between the different texture regions. Our explanation of this result is similar to that made by Regan and Beverley [J. Opt. Soc. Am. 73 (1983) 1684; J. Opt. Soc. Am. A 2 (1985) 147] for simple grating stimuli. However, we show that whereas Regan and Beverley's results could be accounted for on the basis of the tuning functions of the putative mechanisms involved, our results can be explained in terms of the characteristics of the textures themselves. Some implications of our finding are discussed. PMID- 11888537 TI - Minimum displacement thresholds for binocular three-dimensional motion. AB - Thresholds for the detection of motion in depth in the median plane of the head are substantially poorer than those for motion in the frontoparallel plane. This suggests the existence of two independent mechanisms for their detection. Any three-dimensional (3-D) motion can be decomposed into components of motion in the frontoparallel plane and in the median plane of the head. Can human performance for the detection and discrimination of other 3-D motions be predicted by a combination of responses from the two independent mechanisms? Minimum displacement thresholds (d(min)) for the detection of 3-D motion and the direction discrimination of 3-D motion were measured for a wide range of 3-D directions. d(min) data were modelled in terms of the probability summation of a pair of independent motion mechanisms, one responding to motion in the median plane of the head, the second to motion in the frontoparallel plane. Detection of 3-D motion was well predicted by probability summation across a range of 3-D directions. Direction discrimination of 3-D motion was similarly well fit by the probability summation model for multiframe motion displays for some observers. However for two-frame motion displays, direction discrimination for 3-D motion was best fit by a model using only a motion mechanism in the frontoparallel plane. Detection and direction discrimination thresholds for 3-D motion can therefore be explained on the basis of one or two mechanisms, sensitive to motion in the frontoparallel plane and in the median plane of the head. PMID- 11888538 TI - Effect of spatial waveform on apparent spatial frequency. AB - We examined the effect of spatial waveform on the perceived spatial frequency of a grating target. The luminance profile of 0.5 c/ degrees sinusoidal gratings was modified by either compressive or expansive power functions, and was presented alternately with a true sinusoidal grating. Subjects matched the apparent spatial frequency of the two gratings using a method of adjustment. Both compressive and expansive power functions lowered the perceived spatial frequency of the grating, irrespective of the stimulus contrast. Rectified sine wave gratings were also found to reduce apparent spatial frequency. The magnitude of the spatial frequency shifts with spatial waveform diminished with successive matches, which may represent a change in matching strategy employed by observers. Calculations and a further experiment suggest that judgements of spatial frequency may in part be determined by the separation between edges in a grating. PMID- 11888539 TI - Lateral interactions: size does matter. AB - Usually a high-contrast, co-local mask increases contrast threshold (inhibition). Interestingly, a laterally displaced mask (flanker) can facilitate contrast detection (Vision Research 33 (1993) 993; 34 (1994) 73). When spatial scaling of these flanker effects was implied, stimulus bandwidth was confounded with spatial frequency (lambda(-1)). Under conditions where at lower spatial frequencies, the size (standard deviation, sigma) of the Gabor patch was smaller (sigma0.94 at all eccentricities. However, the slope of the function flattened with increasing eccentricity. Using conventional automated perimetry stimuli in perimetrically experienced young subjects, suprathreshold RT increases but threshold RT prolongation decreases with increasing visual field eccentricity. RT fits a power function with decreasing stimulus attenuation but the slope flattens with eccentricity. This relationship found along the nasal horizontal meridian may allow use of RT to cross-check threshold results or to define response windows for reliability indices of conventional automated perimetry. PMID- 11888544 TI - Pictogram naming in dyslexic and normal children assessed by SLO. AB - We measured pictogram naming (PN) and text reading in dyslexic and normally reading young teenagers. Eye movements were monitored by scanning laser ophthalmoscope, revealing positions of fovea, stimuli on the retina, and speech simultaneously. While text reading speed showed the expected difference between groups, PN speeds overlapped widely. PN was mainly controlled by retrieval time in both groups and correlated with age in dyslexics. During PN, only backward saccades occurred more frequently in dyslexics. We conclude that PN activates visual/eidetic mechanisms that are distinct from the phonemic/analytic pathway necessary for reading. This dual organization leads to a wide range of combinations of performances in PN and text reading. PMID- 11888545 TI - Serotonin-based therapeutics. PMID- 11888546 TI - Molecular, pharmacological and functional diversity of 5-HT receptors. AB - Serotonin (5-hydroxytryptamine, 5-HT) is probably unique among the monoamines in that its effects are subserved by as many as 13 distinct heptahelical, G-protein coupled receptors (GPCRs) and one (presumably a family of) ligand-gated ion channel(s). These receptors are divided into seven distinct classes (5-HT(1) to 5 HT(7)) largely on the basis of their structural and operational characteristics. Whilst this degree of physical diversity clearly underscores the physiological importance of serotonin, evidence for an even greater degree of operational diversity continues to emerge. The challenge for modern 5-HT research has therefore been to define more precisely the properties of the systems that make this incredible diversity possible. Much progress in this regard has been made during the last decade with the realisation that serotonin is possibly the least conservative monoamine transmitter and the cloning of its many receptors. Coupled with the actions of an extremely avid and efficient reuptake system, this array of receptor subtypes provides almost limitless signalling capabilities to the extent that one might even question the need for other transmitter systems. However, the complexity of the system appears endless, since posttranslational modifications, such as alternate splicing and RNA editing, increase the number of proteins, oligomerisation and heteromerisation increase the number of complexes, and multiple G-protein suggest receptor trafficking, allowing phenotypic switching and crosstalk within and possibly between receptor families. Whether all these possibilities are used in vivo under physiological or pathological conditions remains to be firmly established, but in essence, such variety will keep the 5-HT community busy for quite some time. Those who may have predicted that molecular biology would largely simplify the life of pharmacologists have missed the point for 5-HT research in particular and, most probably, for many other transmitters. This chapter is an attempt to summarise very briefly 5-HT receptor diversity. The reward for unravelling this complex array of serotonin receptor--effector systems may be substantial, the ultimate prize being the development of important new drugs in a range of disease areas. PMID- 11888547 TI - The medical benefit of 5-HT research. AB - 5-HT research is now more than 50 years old, and it has generated a wealth of therapeutic agents, some of which have had a major impact on disease management. The 5-HT reuptake inhibitors (SSRIs) are among the most widely prescribed drugs for treating depression and a variety of other disorders including anxiety, social phobia and premenstrual dysphoria (PMD). The other major success stories of 5-HT research are the discovery of 5-HT1B/D receptor agonists for treating migraine and 5-HT3 receptor antagonists for chemotherapy and radiation-induced emesis. The role of 5-HT in the mechanism of action of antipsychotic agents remains a topic of intense research, which promises better treatments for schizophrenia in the future. Compounds interacting with 5-HT1F, 5-HT2C, 5-HT6 and 5-HT7 receptors are currently under investigation and may prove to have important therapeutic applications in the future. PMID- 11888548 TI - Subthalamic 5-HT(1A) and 5-HT(1B) receptor modulation of RU 24969-induced behavioral profile in rats. AB - The effects of systemic administration of the serotonin (5-HT)(1A/1B) agonist 5 methoxy-3-(1,2,3,6-tetrahydropyridin-4-yl)1H-indole (RU 24969) on locomotor and investigatory behavior in rats have been well characterized using the behavioral pattern monitor (BPM). To elucidate the neural circuitry underlying this behavioral profile, intracerebral dose--response studies were conducted at two sites with high densities of 5-HT(1B) receptors, the subthalamic nucleus (STN) and substantia nigra. Infusion of RU 24969 into the STN produced systemic RU 24969-like changes in locomotor activity and patterns but an uncharacteristic increase in investigatory holepokes. Intra-STN administration of the selective 5 HT(1A) receptor agonist 8-hydroxy-2-(di-N-propylamino)tetralin (8-OH-DPAT) produced RU 24969-like changes in locomotor patterns only, while the 5-HT(1B) receptor agonist 3(1,2,5,6-tetrahydropyrid-4-yl)pyrrolo[3,2-b]pyrid-5-one dihydrochloride (CP-93,129) increased locomotor activity, produced no change in locomotor patterns and nonsignificantly increased holepokes. Intranigral infusion of RU 24969 produced systemic and intra-STN RU 24969-like increases in locomotor activity. Intranigral RU 24969, however, failed to produce any changes in locomotor patterns or investigatory holepokes. Intranigral infusions of CP-93,129 or 8-OH-DPAT had no effects on locomotor activity, locomotor patterns or investigatory holepokes. These results provide evidence for multiple-site mediation of the locomotor-activating effects of RU 24969 and for a dissociation of the neural substrates underlying locomotor and investigatory components of the RU 24969-induced behavioral profile. PMID- 11888549 TI - Anxiogenic-like effect of serotonin(1B) receptor stimulation in the rat elevated plus-maze. AB - Perturbations in serotonin [5-hydroxytryptamine (5-HT)] neurotransmission have been implicated in several psychiatric illnesses including depression and anxiety disorders. It is not yet clear, however, which of the 14 currently identified 5 HT receptor subtypes in the brain participate in the regulation of emotional states. This study investigates a role for the 5-HT(1B) receptor subtype in anxiety-related behaviors using the elevated plus-maze paradigm in rats. The selective 5-HT(1B) receptor agonist 3-(1,2,5,6-tetrahydro-4-pyridyl)-5 propoxypyrrolo[3,2-b]pyridine (CP 94,253; 1--5.6 mg/kg) dose-dependently decreased the amount of exploration on the open arms of the plus-maze without altering overall locomotor activity. This 5-HT(1B) agonist-induced increase in anxiety-like behavior was dose-dependently reversed by coadministration of the selective 5-HT(1B/1D) receptor antagonist 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol 3-yl)-biphenyl]-amide (GR 127,935). There was no significant effect of GR 127,935 administration alone on plus-maze behavior. These results indicate that 5-HT(1B) receptor activation increases anxiety-like behavioral responses as measured by the elevated plus-maze. Since 5-HT(1B) receptors modulate the activity of multiple neurotransmitter systems that have been implicated in anxiety disorders, these findings suggest that this receptor subtype may represent an important therapeutic target for the treatment of anxiety. PMID- 11888550 TI - Specific labelling of serotonin 5-HT(1B) receptors in rat frontal cortex with the novel, phenylpiperazine derivative, [3H]GR125,743. A pharmacological characterization. AB - Although several tritiated agonists have been used for radiolabelling serotonin (5-hydroxytryptamine, 5-HT)(1B) receptors in rats, data with a selective, radiolabelled antagonist have not been presented. Inasmuch as [3H]GR125,743 specifically labels cloned, human and native guinea pig 5-HT(1B) receptors and has been employed for characterization of cerebral 5-HT(1B) receptor in the latter species [Eur. J. Pharmacol. 327 (1997) 247.], the present study evaluated its utility for characterization of native, cerebral 5-HT(1B) sites in the rat. In homogenates of frontal cortex, [3H]GR125,743 (0.8 nM) showed rapid association (t(1/2)=3.4 min), >90% specific binding and high affinity (K(d)=0.6 nM) for a homogeneous population of receptors with a density (B(max)) of 160 fmol/mg protein. In competition binding studies, affinities were determined for 15 chemically diverse 5-HT(1B) agonists, including 2-[5-[3-(4 methylsulphonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indole-3-yl]ethylamine (L694,247; pK(i), 10.4), 5-carboxamidotryptamine (5-CT; 9.7), 3-[3-(2 dimethylamino-ethyl)-1H-indol-6-yl]-N-(4-methoxybenzyl)acrylamide (GR46,611; 9.6), 5-methoxy-3-(1,2,5,6-tetrahydro-4-pyridinyl)-1H-indole (RU24,969; 9.5), dihydroergotamine (DHE; 8.6), 5-H-pyrrolo[3,2-b]pyridin-5-one,1,4-dihydro-3 (1,2,3,6-tetrahydro-4-pyridinyl (CP93,129; 8.4), anpirtoline (7.9), sumatriptan (7.4), 1-[2-(3-fluorophenyl)ethyl]-4-[3-[5-(1,2,4-triazol-4-yl)-1H-indol-3 yl]propyl]piperazine (L775,606; 6.4) and (minus sign)-1(S)-[2-[4-(4 methoxyphenyl)piperazin-1-yl]ethyl]-N-methyl-3,4-dihydro-1H-2-benzopyran-6 carboxamide (PNU109,291; <5.0). Similarly, affinities were established for 13 chemically diverse antagonists, including N-[4-methoxy-3-(4-methylpiperazin-1 yl)phenyl]-3-methyl-4-(4-pyridyl)benzamide (GR125,743; pK(i), 9.1), ( )cyanopindolol (9.0), (-)-tertatolol (8.2), N-(4-methoxy-3-(4-methylpiperazin-1 yl)phenyl]-2'-methyl-4'-(5-methyl-1,2,4-oxadiozol-3-yl)biphenyl-4-carboxamide (GR127,935; 8.2), N-[3-(1,4-benzodioxan-5-yl)piperidin-4-yl]N-(indan-2yl)amine (S18127; 7.9), metergoline (7.8), (-)-pindolol (7.6), 1'-methyl-5-[2'-methyl-4' (5-methyl-1,2,4-oxadiazol-3-yl)-biphenyl-4-ylcarbonyl]-2,3,6,7-tetrahydro-5H spiro[furo[2,3-f]indole-3,4'-piperidine] (SB224,289; 7.5) and ketanserin (<5.0). These rank orders of affinity correspond to the binding profile of 5-HT(1B) rather than 5-HT(1D) receptors. The low affinities of L775,066 and PNU109,291 versus L694,247 should be noted, as well as the low affinity of ketanserin as compared to SB224,289. Finally, in line with species differences, the affinities of several ligands including CP93,129, RU24,969, (-)-pindolol and (-)-propanolol in rat 5-HT(1B) sites were markedly different to guinea pig 5-HT(1B) sites labelled with [3H]GR125,743. In conclusion, [3H]GR125,743 is an appropriate tool for the radiolabelling of native, rat 5-HT(1B) receptors and permitted determination of the affinities of an extensive series of ligands at these sites. PMID- 11888551 TI - Effect of SB-243213, a selective 5-HT(2C) receptor antagonist, on the rat sleep profile: a comparison to paroxetine. AB - 5-HT(2) receptor antagonists promote slow wave sleep (SWS) in humans and rats, conversely 5-HT(2) agonists inhibit SWS in rats. These alterations are thought to be predominantly mediated via the 5-HT(2C) receptor subtype. It is evident that 5 HT(2) receptor function also plays an important role in depression. Here, we examine the acute effect of the selective 5-HT(2C) receptor antagonist 5-methyl-1 [[-2-[(2-methyl-3-pyridyl)oxy]-5-pyridyl]carbamoyl]-6-triflouromethylindoline hydrochloride (SB-243213-A) on rat sleep in comparison to the selective serotonin reuptake inhibitor (SSRI) paroxetine. Both SB-243213-A (10 mg/kg po) and paroxetine (3 mg/kg po) significantly increased deep SWS (SWS2) quantity (27% and 24%, respectively) and reduced paradoxical sleep (PS) quantity (35%) during the sleep period. Following SB-243213-A, SWS2 occurrence frequency was reduced (24.1%); however, elevated quantity of SWS2 can be attributed to an increase in occurrence duration (81%). Reduced PS quantity results from a decrease in occurrence frequency (46%). In comparison, paroxetine increased SWS2 occurrence frequency (50%), with decreased frequency (27%) and duration (21%) of PS. The data for SB-243213-A in the present study is consistent with that following ritanserin supporting 5-HT(2C) receptor subtype mediation of this response. The similar effect of SB-243213-A to paroxetine with regard to PS quantity provides further evidence that 5-HT(2C) receptor antagonists maybe beneficial in the treatment of depression/anxiety. PMID- 11888552 TI - Biochemical evidence that the atypical antipsychotic drugs clozapine and risperidone block 5-HT(2C) receptors in vivo. AB - Clozapine and risperidone are two atypical antipsychotic drugs which bind, among other receptors, to 5-HT(2C) receptor subtypes. They inhibit the basal inositol phosphate production in mammalian cells expressing rat or human 5-HT(2C) receptors. This biochemical effect is indicative of inverse agonist activity at these receptors. There is evidence that 5-HT(2C) receptors are involved in the control of the activity of central dopaminergic system. Therefore, the effects of clozapine (5 mg/kg ip), risperidone (0.08 mg/kg ip) and of the typical antipsychotic haloperidol (0.1 mg/kg ip) were studied on the extracellular concentration of dopamine (DA) in the nucleus accumbens of chloral hydrate anesthetized rats, using intracerebral microdialysis. When injected alone, clozapine, risperidone and haloperidol caused only small variations in DA efflux. However, clozapine and risperidone completely prevented the inhibitory action of RO 60-0175 (1 mg/kg ip), a 5-HT(2C) receptor agonist, on DA release. On the other hand, haloperidol did not affect RO 60-0175-induced decrease in DA release. Taken together, these data indicate that clozapine and risperidone, unlike haloperidol, are capable of blocking 5-HT(2C) receptors in the nucleus accumbens. It is concluded that the experimental model presented in this study might represent a simple and useful in vivo biochemical method to test the effect of putative atypical antipsychotic drugs on 5-HT(2C) receptors. PMID- 11888553 TI - Influence of the 5-HT2C receptor antagonist, SB-242084, in tests of anxiety. AB - The 5-HT2C antagonist SB-242084 was examined in various anxiety tests at doses based on reversal of mCPP-induced hypoactivity (0.1--3 mg/kg ip). In the elevated plus-maze task, SB-242084 exhibited signs of anxiolysis (time spent, distance travelled, and entries into open arms), but this was potentially confounded by its general increase of locomotion; alprazolam selectively affected open-arm parameters. In a Geller--Seifter conflict test, SB-242084 produced a modest, nonsignificant increase in punished responding compared to the significant effect produced by diazepam. None of the treatments significantly affected unpunished responding. In the conditioned emotional response (CER) test, SB-242084 produced an increase in the suppression ratio (SR, smaller than diazepam). Since this 5 HT2C antagonist also increased lever pressing, an additional test was conducted with amphetamine that stimulated lever pressing but, nonetheless, failed to produce any change in SR. In the fear-potentiated startle task, SB-242084 was inactive in comparison to a significant effect of diazepam. The previously described reduction of schedule-induced polydipsia by fluoxetine and 5-HT2C receptor agonist Ro60-0175 was attenuated by SB-242084 pretreatment, however, the latter compound exhibited a potent increase in polydipsia when given alone. The present results demonstrate an anxiolytic potential of SB-242084, as well as an intrinsic response-enhancing property, however, both of these effects are task dependent. PMID- 11888554 TI - Central 5-HT(4) receptors and dopamine-dependent motor behaviors: searching for a functional role. AB - In this study, we evaluated the role of central 5-HT(4) receptors in the control of motor behaviors related to change of nigrostriatal dopamine (DA) transmission, namely, stereotyped behavior and catalepsy in rats. Indeed, given that 5-HT(4) receptors indirectly modulate nigrostriatal DA neuron activity, we hypothesized that these receptors would regulate nigrostriatal DA transmission in the basal ganglia, and consequently, associated motor responses. Stereotypy was induced either by an acute administration of apomorphine (0.3 and 1.5 mg/kg sc), or by a single morphine administration (15 mg/kg sc) in chronically morphine-treated (15 mg/kg sc, twice daily for 10 days) rats. Catalepsy was induced by the typical neuroleptic haloperidol (HAL; 1 mg/kg sc). The selective 5-HT(4) antagonist, GR 125487 (1 mg/kg ip), modified neither apomorphine- nor morphine-induced stereotypy. HAL-induced catalepsy, while reduced by the systemic administration of the 5-HT(1A) agonist 8-OH-DPAT (0.1 mg/kg sc), was insensitive to GR 125487, systemically (1, 3, 10 mg/kg ip) or locally (20 and 40 nmol/20 microl) administered into the third ventricle. Also, HAL-induced catalepsy was not affected by the selective 5-HT(4) antagonist GR 113808 (3 mg/kg ip). The obtained results indicate that 5-HT(4) receptor antagonism does not modulate motor behaviors related to change of striatal DA transmission. PMID- 11888555 TI - Effects of the 5-HT(6) receptor antagonist, SB-271046, in animal models for schizophrenia. AB - The 5-HT(6) receptor is targeted by several new antipsychotics such as clozapine, olanzapine, and sertindole. We studied the effect of SB-271046 [5-chloro-N-(4 methoxy-3-piperazin-1-yl-phenyl)-3-methyl-2-benzothiophenesulfonamide], a specific 5-HT(6) receptor antagonist, in three models for the positive symptoms of schizophrenia---D-amphetamine-induced hyperactivity, and D-amphetamine- or phencyclidine (PCP)-disrupted prepulse inhibition (PPI). We also tested this compound in a model for the negative symptoms of schizophrenia, PCP-disrupted social interaction (SIT) in rats. Induction of side effects by this compound was evaluated by testing its potency to reduce spontaneous motility, and to induce catalepsy in rats. The effect of SB-271046 was compared to clozapine in all models tested. This study showed that SB-271046 had no beneficial effect in PCP disrupted SIT. However, SB-271046 dose-dependently normalised D-amphetamine disrupted PPI, but did not reverse PCP-disrupted PPI. In addition, SB-271046 did not antagonise D-amphetamine-induced hyperactivity. Thus, this specific 5-HT(6) receptor antagonist was associated with a clear positive outcome in only one model for the positive symptoms of schizophrenia, and had no beneficial effect in the model for negative symptoms. Consequently, it is clear that SB-271046 is not expected to have an antipsychotic efficacy, at least when given as monotherapy. PMID- 11888556 TI - Pharmacological profile of SB-357134: a potent, selective, brain penetrant, and orally active 5-HT(6) receptor antagonist. AB - N-(2,5-Dibromo-3-fluorophenyl)-4-methoxy-3-piperazin-1-ylbenzenesulfonamide (SB 357134) potently inhibited [125I]SB-258585 and [3H]LSD binding in a HeLa cell line expressing human 5-HT(6) receptors (pK(i)=8.6 and 8.54, respectively). Furthermore, SB-357134 inhibited [125I]SB-258585 binding in human caudate- putamen and in rat and pig striatum membranes (pK(i)=8.82, 8.44, and 8.61, respectively). SB-357134 displayed over 200-fold selectivity for the 5-HT(6) receptor versus 72 other receptors and enzymes. 5-HT-stimulated cyclic AMP (cAMP) accumulation in human 5-HT(6) receptors was competitively antagonised by SB 357134 (pA(2)=7.63). SB-357134 inhibited ex vivo [125I]SB-258585 binding in the rat with an ED(50) of 4.9 +/- 1.3 mg/kg po, 4 h postdose. In the rat maximal electroshock seizure threshold (MEST) test, SB-357134 produced a potent and dose dependent increase in seizure threshold, with a minimum effective dose of 0.1 mg/kg po. At 10 mg/kg po, maximum activity occurred between 4 and 6 h postdose. Good exposure was observed with SB-357134 at 10 mg/kg po, reaching maximal blood and brain concentrations of 4.3 +/- 0.2 and 1.3 +/- 0.06 microM, respectively, 1 h postdose. In addition, SB-357134 (10 mg/kg po) enhanced memory and learning following chronic administration (twice a day for 7 days) in the rat water maze. Overall, these studies demonstrate that SB-357134 is a potent, selective, brain penetrant, and orally active 5-HT(6) receptor antagonist. PMID- 11888557 TI - Effects of the 5-HT(7) receptor antagonist SB-258741 in animal models for schizophrenia. AB - The 5-HT(7) receptor is targeted by several new antipsychotics such as clozapine and risperidone. We studied the effect of R-(+)-1-(toluene-3-sulfonyl)-2-[2-(4 methylpiperidin-1-yl)ethyl]-pyrrolidine (SB-258741), a specific 5-HT(7) receptor antagonist, in three models for positive symptoms, D-amphetamine-induced hyperactivity and D-amphetamine- and phencyclidine (PCP)-disrupted prepulse inhibition (PPI) in rats, with the aim of investigating the role of this receptor in the clinical effect of antipsychotics. We also tested this compound in a model for negative symptoms, PCP-disrupted social interaction (SIT) in rats. Induction of side effects by this compound was evaluated by testing its potency to reduce spontaneous motility and to induce catalepsy in rats. The effect of SB-258741 was compared to risperidone in all models. This study showed that SB-258741 had no beneficial effect on PCP-disrupted SIT. SB-258741 did not reverse D-amphetamine disrupted PPI; however, it dose-dependently normalised PCP-disrupted PPI. SB 258741 antagonised D-amphetamine-induced hyperactivity but reduced motility of rats at similar doses. Thus, this specific 5-HT(7) receptor antagonist brought a clear positive outcome in only one model for positive symptoms of schizophrenia and had no beneficial effect in the model for negative symptoms. Consequently, it is clear that SB-258741 affects the PPI phenomenon but is not expected to have an antipsychotic effect on its own in clinic. PMID- 11888558 TI - Induction of hyperlocomotion in mice exposed to a novel environment by inhibition of serotonin reuptake. A pharmacological characterization of diverse classes of antidepressant agents. AB - This study characterized the influence of acute administration of diverse classes of antidepressant agent upon the spontaneous locomotor activity (LA) of mice in a novel, open-field environment. The selective serotonin (5-HT) reuptake inhibitors (SSRIs), citalopram, fluoxetine, paroxetine, fluvoxamine, litoxetine and zimelidine, dose-dependently enhanced LA. Their actions were mimicked by the mixed 5-HT/noradrenaline (NA) reuptake inhibitors (SNRIs), venlafaxine, duloxetine and S33005. In contrast, clomipramine only slightly elevated LA and two further tricyclics, imipramine and amitriptyline, were inactive. Further, the selective NA vs. 5-HT reuptake inhibitors (NARIs), reboxetine, desipramine, maprotiline, nisoxetine and nortriptyline all failed to increase LA. The "atypical antidepressants," mianserin and mirtazapine, neither of which modify 5 HT reuptake, as well as the mixed SSRI/5-HT(2) antagonists, nefazodone and trazodone, also failed to increase LA. Doses of SSRI and SNRI which increased LA did not modify motor performance in the rotarod test. Further, they did not enhance LA in rats, suggesting that this response is characteristic of mice. Finally, upon prehabituation of mice to the activity chamber, the SSRI, citalopram, and the SNRI, venlafaxine, failed to increase LA. In conclusion, in mice exposed to a novel environment, inhibition of 5-HT reuptake by SSRIs and SNRIs enhances spontaneous LA in the absence of a generalized influence upon motor function. This response provides a simple parameter for characterization of SSRIs and SNRIs, and differentiates them from other classes of antidepressant agent. Although an influence upon arousal and/or anxiety is likely related to the increase in LA, the functional significance of this response requires additional elucidation. PMID- 11888559 TI - Parabrachial infusion of D-fenfluramine reduces food intake. Blockade by the 5 HT(1B) antagonist SB-216641. AB - Systemic administration of the serotonin (5-HT) releaser/reuptake inhibitor, D fenfluramine decreases consumption of food in mammals. This hypophagic action involves loci at several levels of the neuraxis. Indirect evidence implicates the parabrachial nucleus (PBN) of the pons as one of these regions. Consistent with this hypothesis, unilateral infusion of D-fenfluramine (200, 280, and 400 nmol/0.5 microl) directly into the lateral PBN (LPBN) of male rats reduced food intake by 33%, 56%, and 66% from baseline (7.3 +/- 0.7 g) during a 30-min test with chow. Infusions lateral, medial, and dorsal to the PBN were ineffective. Stimulating 5-HT(1B) receptors in the PBN also reduces feeding. Administration of the selective 5-HT(1B) agonist CP-93,129 (3-(1,2,5,6-tetrahydropyrid-4 yl)pyrrolo[3,2-b]pyrid-5-one) (0, 0.625, 2.5, and 10 nmol/0.5 microl) into the PBN reduced food intake by 25--79%. The selective 5-HT(1B) antagonist SB-216641 (N-[3-[3-(dimethylamino(ethoxy]-4-methoxyphenyl]-2'-methyl-4'-(5-methyl-1,2,4 oxadiazol-3-yl)-[1,1'-biphenyl]-4-carboxamide) (2.5 nmol) completely blocked the hypophagic action of the approximate ED(50) doses of CP-93,129 (2.5 nmol) and D fenfluramine (280 nmol). These data strongly suggest that directly or indirectly activating 5-HT(1B) receptors in the LPBN inhibits feeding and implicates this pontine region in the serotonergic regulation of eating and satiation. PMID- 11888560 TI - Serotonin 2C receptor agonists and the behavioural satiety sequence in mice. AB - The studies reported here examined the role of the 5-hydroxytryptamine (5-HT)(2C) receptor subtype in the control of ingestive behaviour in mice. Behavioural satiety sequence (BSS) and food intake measurements were taken, comparing the selective 5-HT(2C) receptor agonist (S)-2-(6-chloro-5-fluoro-indol-l-yl)-l methylethylamine hydrochloride (Ro 60-0175; 1.0, 3.0 and 10.0 mg/kg) and D fenfluramine (3.0 mg/kg). Ro 60-0175 produced a dose-dependent decrease in food intake. The effects of Ro 60-0175 (3.0 mg/kg) on the BSS were similar to the hypophagic effects of D-fenfluramine (3.0 mg/kg). In a second experiment, the specific effects on feeding produced by Ro 60-0175 (5.6 mg/kg) were attenuated by pretreatment with the selective 5-HT(2C) receptor antagonist 6-chloro-5-methyl-1 [2(2-methylpyridyl-3-oxy)-pyrid-5-yl carbamoyl] indoline (SB 242084; 0.5 mg/kg). The 5-HT(1B/2C) receptor agonist 1-(m-chlorophenyl)piperazine (mCPP; 3 mg/kg) also produced a substantial decrease in food intake, which was attenuated by SB 242084 (0.5 mg/kg). A dose of the selective 5-HT(1B/1D) antagonist 2'-methyl-4'(5 methyl-[1,2,4]oxadiazol-3-yl)-biphenyl-4-carboxylic acid [4-(5-methoxy-3-(4 methyl-piperazin-1-yl)-phenyl]amide (GR 127935; 3.0 mg/kg) that successfully attenuated the action of the 5-HT(1B) agonist 5-methoxy-3(1,2,3,6 tetrahydropyridin-4-yl)-1H-indole (RU 24969; 5.0 mg/kg) failed to attenuate mCPP induced hypophagia. These data suggest that Ro 60-0175- and mCPP-induced hypophagia in mice are mediated via activation of 5-HT(2C) receptors and that stimulation of 5-HT(1B) receptors plays only a minor role in mCPP-induced hypophagia. PMID- 11888561 TI - Estrous cycle and food availability affect feeding induced by amygdala 5-HT receptor blockade. AB - We have recently reported that bilateral infusions of the 5-HT receptor antagonist metergoline (MET) into the posterior basolateral amygdala (pBLA) elicit feeding in female rats tested at mid-light cycle. The present study was performed to determine whether (1) testing at two different phases of the estrous cycle, and/or (2) the palatability of the food might modify this effect. Subjects were 18 adult females with bilateral pBLA cannulae. Following familiarization with Froot Loops cereal, a within-subjects design tested all animals for 1- and 2 h food intake under 2 Drug (0.3 nmol MET vs. Vehicle), 2 Estrous Cycle (diestrus vs. estrus) and 2 Food (lab chow vs. Froot Loops) conditions. Rats weighed more at diestrus than at proestrus (P<.05) or estrus (P<.005). Multivariate analyses of variance (MANOVAs) revealed a preference for Froot Loops over lab chow (P<.0001). MET increased feeding regardless of food type (P<.0001). Rats ate more Froot Loops (P<.01), but not lab chow, at diestrus vs. estrus. A three-way interaction (P<.05) showed rats ate more during the first hour in estrus than in diestrus to lab chow but not Froot Loops. These data suggest pBLA MET differentially affects feeding over the estrous cycle depending on the palatability of food available. PMID- 11888562 TI - Hypothalamic paraventricular 5-hydroxytryptamine: receptor-specific inhibition of NPY-stimulated eating and energy metabolism. AB - The feeding effects of 5-hydroxytryptamine (5-HT)(1) and 5-HT(2) receptor agonists injected into the hypothalamic paraventricular nucleus (PVN) immediately prior to PVN administration of neuropeptide Y (NPY) were examined. The impact of these same compounds on NPY-induced alterations in energy metabolism was also assessed in an attempt to characterize further the potential interactive relationship of PVN NPY and 5-HT on feeding and whole body calorimetry. Specifically, several experiments examined the effect of various 5-HT receptor agonists on NPY-stimulated eating and alterations in energy substrate utilization [respiratory quotient (RQ)]. This included the 5-HT(1A) receptor agonist 8-OH DPAT, the 5-HT(1B/1A) agonist RU 24969, the 5-HT(1D) agonist L-694,247, the 5 HT(2A/2C) agonist DOI, the 5-HT(2B) agonist BW 723C86 and the 5-HT(2C) agonist mCPP. In feeding tests conducted at the onset of the dark cycle, drugs were administered 5 min prior to PVN injection of NPY and food intake was measured 2 h postinjection. The metabolic effects of NPY following a similar pretreatment were monitored using an open-circuit calorimeter measuring the volume of oxygen consumed (VO(2)), carbon dioxide produced (VCO(2)) and RQ (VCO(2)/VO(2)). PVN injection of NPY (100 pmol) potentiated feeding and evoked reliable increases in RQ. Only DOI (2.5--5 nmol) pretreatment antagonized NPY-induced eating and blocked the peptide's effect on energy substrate utilization. Direct PVN pretreatment with spiperone (SPRN), a 5-HT(2A) receptor antagonist, and ketanserin (KTSN), a 5-HT(2A/2C) antagonist, but not SDZ SER 082, a 5-HT(2B/2C) antagonist, or the 5-HT(2C) antagonist RS 102221, blocked the effect of DOI in both feeding and metabolic tests providing additional evidence that activation of PVN 5-HT(2A) receptors inhibits NPY's action on feeding and substrate utilization. PMID- 11888564 TI - Role of serotonin(2C) receptors in the control of brain dopaminergic function. AB - There is substantial evidence that the functional status of mesocorticolimbic dopaminergic (DA) system originating in the ventral tegmental area (VTA) is under a phasic and tonic inhibitory control by the serotonergic system, which acts by stimulating serotonin(2C) (5-HT(2C)) receptor subtypes. This assertion is based upon a number of electrophysiological and biochemical data showing that 5-HT(2C) receptor agonists decrease, while 5-HT(2C) receptor antagonists enhance mesocorticolimbic DA function. On the other hand, it does not seem that 5-HT(2C) receptors play a relevant role in the control of nigrostriatal DA system originating in the substantia nigra pars compacta (SNc). The authors of this article review the most relevant data regarding the role of 5-HT(2C) receptors in the control of brain DA function and underline the importance of this subject in the search of new therapies for neuropsychiatric disorders such as depression, schizophrenia, drug addiction, and Parkinson's disease. PMID- 11888563 TI - Activation of 5-HT(1B) receptors in the nucleus accumbens reduces self administration of amphetamine on a progressive ratio schedule. AB - Brain serotonin interacts with dopamine function in a complex fashion. Previous work from our laboratory showed that activation of 5-HT(1B) receptors within the nucleus accumbens attenuates the ability of amphetamine to increase responding for conditioned reinforcement. The primary purpose of these experiments was to determine the impact of 5-HT receptor stimulation, with particular focus on 5 HT(1B) receptors in the nucleus accumbens on the reinforcing effect of amphetamine. To this end several experiments determined the effects of injecting 5-HT, and various 5-HT agonists, into the nucleus accumbens on responding for intravenous infusions of amphetamine (60 microg/kg) delivered according to a progressive ratio schedule of reinforcement. Both 5-HT (2.5, 5 and 10 microg) and the selective 5-HT(1B) receptor agonist CP93,129 (0.625, 1.25 and 2.5 microg) dose-dependently reduced responding for amphetamine. Injections of 5-HT but not CP93,129 also reduced responding for food under a similar PR schedule. The 5 HT(1A) agonist 8-OH-DPAT (5 microg) and the nonselective 5-HT(2) agonist DOI (10 microg) failed to alter amphetamine self-administration. Pretreatment with the selective 5-HT(1B/1D) receptor antagonist GR127935 (3 mg/kg) attenuated the ability of 5-HT and CP93,129 to reduce amphetamine self-administration following their injection into the nucleus accumbens. These results extend our previous findings that increasing 5-HT activity in the nucleus accumbens inhibits dopamine dependent behaviour, and further indicate that activation of 5-HT(1B) receptors is particularly important in this regard. PMID- 11888565 TI - An investigation of the role of 5-HT(2C) receptors in modifying ethanol self administration behaviour. AB - We have previously reported that the 5-HT uptake blocker and releaser, dexfenfluramine, attenuates ethanol intake, and that this may be mediated via a 5 HT(2C) receptor mechanism. Our goals were to further determine the contribution made by this receptor subtype in mediating the reduction in ethanol self administration induced by dexfenfluramine using the selective 5-HT(2C) antagonist, SB242,084. Additionally, we wanted to compare dexfenfluramine's effects on ethanol motivated responding with those elicited by the 5-HT(2C) receptor agonist Ro60-0175. In male Wistar rats trained to self-administer a 12% w/v ethanol solution on an FR-4 schedule, both dexfenfluramine (0.05--2.5 mg/kg ip) and Ro60-0175 (0.1--1 mg/kg sc) produced a significant dose-dependent reduction in ethanol self-administration, which was reversed by SB242,084 (0.5 mg/kg ip). Interestingly, SB242,084 alone (0.1--1 mg/kg ip) significantly increased ethanol motivated responding in both high and low ethanol drinking animals. While dexfenfluramine had no effect on ethanol's kinetic profile, the selective 5-HT(2C) agents used had opposing effects, with the agonist Ro60-0175 decreasing and the antagonist SB242,084 increasing blood ethanol levels. Since there were incongruent drug effects on ethanol self-administration and blood ethanol levels, these data support a role for 5-HT(2C) receptors in modifying ethanol intake independent of their effects on blood ethanol kinetics. Furthermore, 5-HT(2C) receptors may exert a tonic control over ethanol self administration behaviour, since agonist and antagonist administration had opposing effects on this behaviour. PMID- 11888566 TI - Serotonin 5-HT(2C) receptors in nucleus accumbens regulate expression of the hyperlocomotive and discriminative stimulus effects of cocaine. AB - The serotonin 5-HT(2C) receptor (5-HT(2C)R) is abundant in the nucleus accumbens (NAc) shell and is considered an important target for 5-HT to modulate the dopamine (DA) mesoaccumbens circuit, which plays a prominent role in the behavioral effects of cocaine. The present study analyzed the ability of intra NAc shell infusions of the 5-HT(2C)R agonists, MK 212 and RO 60-0175, or the 5 HT(2C)R antagonist, RS 102221, to alter either spontaneous or cocaine-evoked activity as well as the discriminative stimulus properties of cocaine. In male Sprague--Dawley rats implanted with bilateral cannulae aimed at the NAc shell, locally injected MK 212 (0.05--0.5 microg/side) or RO 60-0175 (0.5--5 microg/side) did not alter spontaneous activity, but dose-dependently enhanced hyperactivity evoked by cocaine (10 mg/kg ip). In rats trained to discriminate cocaine (10 mg/kg ip) from saline (ip) in a two-lever, water-reinforced FR 20 task, intra-NAc microinfusion of MK 212 (0.05 microg/side) or RO 60-0175 (0.5 microg/side) evoked 37% or 48% cocaine lever responding, respectively. Both MK 212 (0.05 microg/side) and RO 60-0175 (0.5 microg/side) enhanced the discriminability of submaximal doses of cocaine (0.625--2.5 mg/kg). Moreover, intra-NAc infusion of RS 102221 (0.05--1.5 microg/side) dose-dependently attenuated the stimulus effects of cocaine. These data reinforce the hypothesis that 5-HT(2C)R plays a role in the regulatory neurochemistry of the NAc shell that is important to the full expression of the behaviors evoked by cocaine. PMID- 11888567 TI - Involvement of serotonin in nicotine dependence: processes relevant to positive and negative regulation of drug intake. AB - The neurobiological substrate of nicotine dependence has been the subject of extensive preclinical and clinical research. Many experimental reports have implicated the brain serotonin (5-HT) systems in processes relevant to nicotine dependence, but the specific role of this neurotransmitter system largely remains to be elucidated. This review will focus on the role of 5-HT in the acute and chronic effects of nicotine. In particular, the evidence for a role of 5-HT neurotransmission in brain processes thought to be involved in positive and negative control of nicotine use will be examined, and potential clinical implications discussed. PMID- 11888568 TI - 5-Hydroxytryptamine and interval timing behaviour. AB - Interval timing behaviour is revealed by prospective, immediate and retrospective timing schedules. Prospective timing tasks are used to study intertemporal choice (choice between outcomes occurring after different delays), immediate timing tasks to study temporal differentiation (temporal regulation of the animal's behaviour) and retrospective timing tasks to study temporal discrimination (discrimination between the durations of external events). Central 5 hydroxytryptamine (5-HT) depletion promotes preference for small early reinforcers over large delayed reinforcers, possibly by facilitating the time dependent degradation of reinforcer value. Central 5-HT depletion retards the learning of temporal differentiation, and increases the variability of timing in some immediate timing tasks; however, it does not impede (in some cases it facilitates) the acquisition of temporal discrimination. Attempts to ascribe all the effects of 5-HT depletion on timing to a single behavioural process have been unsuccessful, although disinhibition of switching between operant responses may account for some of the findings. Acute treatment with drugs affecting 5-HTergic mechanisms alters timing behaviour in qualitatively different ways in different timing schedules, casting doubt on the idea that the effects of these drugs are mediated by interaction with a unitary timing process. The receptors that mediate 5-HT's putative involvement in interval timing behaviour remain to be identified. PMID- 11888569 TI - Effect of 8-OH-DPAT on temporal discrimination following central 5 hydroxytryptamine depletion. AB - The 5-hydroxytryptamine (5-HT)(1A) receptor agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) alters performance in discrete-trials timing schedules. 5-HT(1A) receptors occur both presynaptically and postsynaptically, but it is not known which receptor population mediates the effects of 8-OH-DPAT on timing. Rats received intra-raphe injections of 5,7-dihydroxytryptamine (n=16) or sham lesions (n=14). They were trained in a discrete-trials psychophysical procedure in which levers were presented at a predetermined time after the onset of each trial (2.5, 7.5,., 47.5 s). A response on lever A was reinforced if lever presentation occurred < 25 s after trial onset; a response on lever B was reinforced if lever presentation occurred >25 s after trial onset. After 70 preliminary sessions, the rats received 8-OH-DPAT (25, 50, 100, 200 microg kg(-1) sc) and saline vehicle. The percentage of responses on lever B (%B) increased as a function of time from trial onset. Under the baseline (vehicle-treatment) condition, performance did not differ between the two groups. 8-OH-DPAT did not alter the indifference point (time corresponding to %B=50%), but dose-dependently increased the Weber fraction in both groups. Forebrain concentrations of 5-HT and 5-HIAA in the lesioned group were approximately 10% of control levels. The results suggest that the effect of 8-OH-DPAT on performance on discrete-trials timing schedules is mediated by postsynaptic 5-HT(1A) receptors. PMID- 11888570 TI - Nicotinic--serotonergic interactions in brain and behaviour. AB - This review focuses on nicotinic--serotonergic interactions in the central nervous system (CNS). Nicotine increases 5-hydroxytryptamine (5-HT) release in the cortex, striatum, hippocampus, dorsal raphe nucleus (DRN), hypothalamus, and spinal cord. As yet, there is little firm evidence for nicotinic receptors on serotonergic terminals and thus nicotine's effects on 5-HT may not necessarily be directly mediated, but there is strong evidence that the 5-HT tone plays a permissive role in nicotine's effects. The effects in the cortex, hippocampus, and DRN involve stimulation of 5-HT(1A) receptors, and in the striatum, 5-HT(3) receptors. The 5-HT(1A) receptors in the DRN play a role in mediating the anxiolytic effects of nicotine and the 5-HT(1A) receptors in the dorsal hippocampus and lateral septum mediate its anxiogenic effects. The increased startle and anxiety during nicotine withdrawal is mediated by 5-HT(1A) and 5 HT(3) receptors. The locomotor stimulant effect of acute nicotine is mediated by 5-HT(1A) receptors and 5-HT(2) receptors may play a role in the expression of a sensitised response after chronic nicotine treatment. Unfortunately, the role of 5-HT(1A) receptors in mediating nicotine seeking has not yet been investigated and would seem an important area for future research. There is also evidence for nicotinic--serotonergic interactions in the acquisition of the water maze, passive avoidance, and impulsivity in the five-choice serial reaction task. PMID- 11888571 TI - Electrical stimulation of the dorsal and median raphe nuclei increases extracellular noradrenaline in rat hippocampus: Evidence for a 5-HT-independent mechanism. AB - Recent studies have used raphe stimulation combined with in vivo measurements of extracellular dopamine to investigate interactions between the 5 hydroxytryptamine (5-HT) and dopamine systems. Here we have tested whether the same approach can be used to investigate interactions between the 5-HT and noradrenaline systems. Electrical stimulation of the dorsal raphe nucleus (DRN) or median raphe nucleus (MRN) was performed in anaesthetised rats implanted with microdialysis probes in the hippocampus and locus coeruleus (LC). DRN stimulation (3, 5 and 10 Hz) evoked a frequency-dependent increase in extracellular noradrenaline in the hippocampus. MRN stimulation had a similar effect. Both DRN and MRN stimulations enhanced extracellular 5-HT levels in the LC and previous studies have demonstrated that extracellular 5-HT also increases in the hippocampus. However, the increase in hippocampal noradrenaline evoked by DRN stimulation was not altered by 5-HT neuronal lesions, which reduced 5-HT metabolite levels by 90%. In conclusion, electrical stimulation of the midbrain raphe increases extracellular noradrenaline in the hippocampus, however, experiments in 5-HT-lesioned animals suggest that this response is not mediated by 5-HT. Although raphe stimulation may be useful to investigate interactions between 5-HT and dopamine, our data indicate that the same approach may not be feasible for 5-HT and noradrenaline. PMID- 11888572 TI - Alteration in 5-hydroxytryptamine agonist-induced behaviour following a corticosterone implant in adult rats. AB - Hypercortisolism and altered serotonergic function may account for the pathological symptoms seen in depression. This study examines the impact of 4 days continuous corticosterone treatment on 5-HT agonist-induced behaviour to delineate changes in 5-HT receptor function in the adult rat. The flat body posture, reciprocal forepaw treading, elevated corticosterone, hyperglycaemia, hypothermia and reduced hippocampal 5-HT induced by the 5-HT(1A) agonist 8-OHDPAT (0.3 mg/kg ip) were all significantly attenuated by the corticosterone implant. The elevation in plasma corticosterone and back muscle contractions evoked by the 5-HT(2A) agonist DOI (1 mg/kg ip) were attenuated, whilst wet-dog shakes were enhanced by corticosterone treatment. 5-HT(2B) agonist-induced behaviour and the hypolocomotion and hypophagia induced by the 5-HT(2C) agonist m-CPP (2.5 mg/kg ip) were unaltered but the mCPP-induced elevation in corticosterone was abolished by corticosterone treatment. Hypothalamic 5-HT receptors mediating corticosterone and 5-HT(1A) receptors, whether on serotonergic nerve terminals or postsynaptic neurones, were downregulated by corticosterone treatment. In contrast, 5-HT(2A) receptors may be up- or downregulated dependent on whether they are on supraspinal or spinal neurones, respectively. A comparison of the brain region dependent alteration in serotonergic function produced by hypercorticosterone in the rat with that seen in depression is discussed. PMID- 11888573 TI - Serotonin releasing agents. Neurochemical, therapeutic and adverse effects. AB - This review summarizes the neurochemical, therapeutic and adverse effects of serotonin (5-HT) releasing agents. The 5-HT releaser (plus minus)-fenfluramine is composed of two stereoisomers, (+)-fenfluramine and (minus sign)-fenfluramine, which are N-de-ethylated to yield the metabolites, (+)-norfenfluramine and (minus sign)-norfenfluramine. Fenfluramines and norfenfluramines are 5-HT transporter substrates and potent 5-HT releasers. Other 5-HT releasing agents include m chlorophenylpiperazine (mCPP), a major metabolite of the antidepressant drug trazodone. Findings from in vitro and in vivo studies support the hypothesis that fenfluramines and mCPP release neuronal 5-HT via a non-exocytotic carrier mediated exchange mechanism involving 5-HT transporters. (+)-Norfenfluramine is a potent 5-HT(2B) and 5-HT(2C) receptor agonist. The former activity may increase the risk of developing valvular heart disease (VHD), whereas the latter activity is implicated in the anorectic effect of systemic fenfluramine. Anorectic agents that increase the risk of developing primary pulmonary hypertension (PPH) share the common property of being 5-HT transporter substrates. However, these drugs vary considerably in their propensity to increase the risk of PPH. In this regard, neither trazodone nor mCPP is associated with PPH. Similarly, although some 5-HT substrates can deplete brain 5-HT (fenfluramine), others do not (mCPP). In addition to the established indication of obesity, 5-HT releasers may be helpful in treating psychiatric problems such as drug and alcohol dependence, depression and premenstrual syndrome. Viewed collectively, it seems possible to develop new medications that selectively release 5-HT without the adverse effects of PPH, VHD or neurotoxicity. Such agents may have utility in treating a variety of psychiatric disorders. PMID- 11888574 TI - Recreational Ecstasy/MDMA, the serotonin syndrome, and serotonergic neurotoxicity. AB - The ring-substituted amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA) or "Ecstasy" is widely used a recreational drug. It stimulates the release and inhibits the reuptake of serotonin (5-HT) and other neurotransmitters such as dopamine to a lesser extent. The acute boost in monoamine activity can generate feelings of elation, emotional closeness, and sensory pleasure. In the hot and crowded conditions of raves/dances, mild versions of the serotonin syndrome often develop, when hyperthermia, mental confusion, and hyperkinesia predominate. Rest in a cooler environment generally reverses these problems, although they can develop into medical emergencies, which occasionally prove fatal. This acute serotonergic overactivity is exacerbated by the high ambient temperatures, overcrowding (aggregate toxicity), and use of other stimulant drugs. The on-drug experience is generally followed by negative moods, with 80--90% of weekend Ecstasy users reporting 'midweek blues', due probably to monoaminergic depletion. Single doses of MDMA can cause serotonergic nerve damage in laboratory animals, with repeated doses causing extensive loss of distal axon terminals. Huether's explanatory model for this 5-HT neurotoxicity will be briefly described. There is an increasing body of evidence for equivalent neuropsychobiological damage in humans. Abstinent regular Ecstasy users often show: reduced cerebrospinal 5-HIAA, reduced density of 5-HT transporters, blunted response to a fenfluramine challenge, memory problems, higher cognitive deficits, various psychiatric disorders, altered appetite, and loss of sexual interest. Functional deficits may remain long after drug use has ceased and are consistent with serotonergic axonal loss in higher brain regions. PMID- 11888575 TI - How strong is the evidence that brain serotonin neurons are damaged in human users of ecstasy? AB - Understanding the diverse functions of serotonin in the human brain can be obtained through examination of subjects having a lower than normal number of brain serotonin neurons. Behavioral abnormalities consistent with brain serotonergic damage have been reported in some polydrug users who also use the neurotoxin ecstasy (methylenedioxymethamphetamine, MDMA). This review evaluates the evidence from neuroimaging studies that brain serotonergic damage is a feature of human users of ecstasy. To date, neuroimaging studies designed to establish whether levels of brain serotonin neurons are lower than normal in ecstasy users have employed radioligands that bind to one component of the serotonin neuron, the serotonin transporter (SERT). Because these studies are methodologically flawed in terms of reliability or validity of the SERT measurement and appear to have employed polydrug users, no definitive information is yet available on the question of ecstasy toxicity to human brain serotonin neurons. Until these issues are resolved, it cannot be assumed that ecstasy exposure represents a chronic serotonin deficiency condition. PMID- 11888576 TI - The role of serotonin in human mood and social interaction. Insight from altered tryptophan levels. AB - Alterations in brain tryptophan levels cause changes in brain serotonin synthesis, and this has been used to study the implication of altered serotonin levels in humans. In the acute tryptophan depletion (ATD) technique, subjects ingest a mixture of amino acids devoid of tryptophan. This results in a transient decline in tissue tryptophan and in brain serotonin. ATD can result in lower mood and increase in irritability or aggressive responding. The magnitude of the effect varies greatly depending on the susceptibility of the subject to lowered mood or aggressivity. Unlike ATD, tryptophan can be given chronically. Tryptophan is an antidepressant in mild to moderate depression and a small body of data suggests that it can also decrease aggression. Preliminary data indicate that tryptophan also increases dominant behavior during social interactions. Overall, studies manipulating tryptophan levels support the idea that low serotonin can predispose subjects to mood and impulse control disorders. Higher levels of serotonin may help to promote more constructive social interactions by decreasing aggression and increasing dominance. PMID- 11888577 TI - A backhanded assault on academic freedom. PMID- 11888578 TI - Individualising HIV treatment--pharmacogenetics and immunogenetics. PMID- 11888579 TI - Infections in neutropenic cancer patients. PMID- 11888580 TI - Graduated licensing programmes and young-driver crashes. PMID- 11888581 TI - An unhappy coincidence between multiple sclerosis and trauma? PMID- 11888582 TI - Association between presence of HLA-B*5701, HLA-DR7, and HLA-DQ3 and hypersensitivity to HIV-1 reverse-transcriptase inhibitor abacavir. AB - BACKGROUND: The use of abacavir--a potent HIV-1 nucleoside-analogue reverse transcriptase inhibitor--is complicated by a potentially life-threatening hypersensitivity syndrome in about 5% of cases. Genetic factors influencing the immune response to abacavir might confer susceptibility. We aimed to find associations between MHC alleles and abacavir hypersensitivity in HIV-1-positive individuals treated with abacavir. METHODS: MHC region typing was done in the first 200 Western Australian HIV Cohort Study participants exposed to abacavir. Definite abacavir hypersensitivity was identified in 18 cases, and was excluded in 167 individuals with more than 6 weeks' exposure to the drug (abacavir tolerant). 15 individuals experienced some symptoms but did not meet criteria for abacavir hypersensitivity. p values were corrected for comparisons of multiple HLA alleles (p(c)) by multiplication of the raw p value by the estimated number of HLA alleles present within the loci examined. FINDINGS: HLA-B*5701 was present in 14 (78%) of the 18 patients with abacavir hypersensitivity, and in four (2%) of the 167 abacavir tolerant patients (odds ratio 117 [95% CI 29-481], p(c)<0.0001), and the HLA-DR7 and HLA-DQ3 combination was found in 13 (72%) of hypersensitive and five (3%) of tolerant patients (73 [20-268], p(c)<0.0001 ). HLA-B*5701, HLA-DR7, and HLA-DQ3 were present in combination in 13 (72%) hypersensitive patients and none of the tolerant patients (822 [43-15 675], p(c)<0.0001). Other MHC markers also present on the 57.1 ancestral haplotype to which the three markers above belong confirmed the presence of haplotype-specific linkage disequilibrium, and mapped potential susceptibility loci to a region bounded by C4A6 and HLA-C. Within the entire abacavir-exposed cohort (n=200), presence of HLA-B*5701, HLA-DR7, and HLA-DQ3 had a positive predictive value for hypersensitivity of 100%, and a negative predictive value of 97%. INTERPRETATION: Genetic susceptibility to abacavir hypersensitivity is carried on the 57.1 ancestral haplotype. In our population, withholding abacavir in those with HLA B*5701, HLA-DR7, and HLA-DQ3 should reduce the prevalence of hypersensitivity from 9% to 2.5% without inappropriately denying abacavir to any patient. PMID- 11888583 TI - Comparison of dual nucleoside-analogue reverse-transcriptase inhibitor regimens with and without nelfinavir in children with HIV-1 who have not previously been treated: the PENTA 5 randomised trial. AB - INTRODUCTION: Treatment options for children with HIV-1 are limited. We aimed to compare activity and safety of three dual-nucleoside analogue reverse transcriptase inhibitor (NRTI) regimens with or without a protease inhibitor in previously untreated children with HIV-1. METHODS: In our multicentre trial, we randomly assigned 36 children to zidovudine and lamivudine, 45 to zidovudine and abacavir, and 47 to lamivudine and abacavir. Children who were symptom-free (n=55) were also randomly assigned to receive nelfinavir or placebo. Children with more advanced disease received open-label nelfinavir (73). Primary endpoints were change in plasma HIV-1 RNA at 24 and 48 weeks for the NRTI comparison and occurrence of serious adverse events for both randomised comparisons. Analyses were by intention to treat. FINDINGS: Children had a median CD4 percentage of 22% (IQR 15-29) and a mean HIV-1 RNA concentration of 5.0 log copies/mL (SD 0.8). One child was lost to follow-up and one died of sepsis. At 48 weeks, in the zidovudine/lamivudine, zidovudine/abacavir, and lamivudine/abacavir groups, mean HIV-1 RNA had decreased by 1.71, 2.19, and 2.63 log copies/mL, respectively (estimated in absence of nelfinavir) (p=0.02 after adjustment for baseline factors). One child had a hypersensitivity reaction to abacavir; and three with possible reactions stopped abacavir. There were 24 serious adverse events--six in the symptom-free children (all on nelfinavir), but none were attributed to nelfinavir. INTERPRETATION: Regimens containing abacavir were more effective than zidovudine/lamivudine. Such regimens could be combined with protease inhibitors and non-nucleoside reverse transcriptase inhibitors for safe and effective treatment of previously untreated children with HIV-1. PMID- 11888584 TI - Efficacy and safety of tacrolimus compared with ciclosporin microemulsion in renal transplantation: a randomised multicentre study. AB - BACKGROUND: In previous comparative studies tacrolimus was superior to the standard formulation of ciclosporin in preventing acute rejection after renal transplantation. We have compared the microemulsion formulation of ciclosporin with tacrolimus in a multicentre randomised trial. METHODS: The 6-month open study involved 560 patients in 50 European centres. 287 patients were randomly assigned tacrolimus and 273 ciclosporin microemulsion plus azathioprine and corticosteroids. The initial oral daily doses were 0.30 mg/kg for tacrolimus and 8-10 mg/kg for ciclosporin. The primary endpoint was the proportion of patients with biopsy-proven acute rejection and the time to this event. FINDINGS: The two study groups were similar in terms of baseline characteristics. Three patients did not receive study treatment or did not undergo transplantation (one tacrolimus, two ciclosporin). The rate of biopsy-confirmed acute rejection was significantly lower with tacrolimus than with ciclosporin microemulsion (56 patients [19.6%] vs 101 [37.3%]; 17.7% difference [95% CI 10.3-25.1]; p<0.0001). Biopsy-confirmed corticosteroid-resistant rejection was also significantly lower with tacrolimus (27 [9.4%] vs 57 [21.0%]; 11.6% difference [5.7-17.5]; p<0.0001). Cross-over between therapies because of biopsy-proven rejection was judged necessary in one of 286 (0.3%) tacrolimus-group patients and 27 of 271 (10.0%) ciclosporin-group patients (p<0.0001). There were no significant differences in survival of patients or grafts or in renal function. The overall frequency of adverse events was similar in the two groups, though hypertension and hypercholesterolaemia were more common in the ciclosporin group and tremor and hypomagnesaemia were more frequent in the tacrolimus group. INTERPRETATION: Tacrolimus was significantly more effective than ciclosporin microemulsion in preventing acute rejection after renal transplantation and had a superior cardiovascular-risk profile. PMID- 11888585 TI - Red blood cell methylfolate and plasma homocysteine as risk factors for venous thromboembolism: a matched case-control study. AB - BACKGROUND: Moderate hyperhomocysteinaemia is a risk factor for venous thromboembolism. We do not know whether this risk depends on homocysteine itself or on components of the homocysteine remethylation pathway, such as methylfolate. We did a case-control study to analyse the relation between the major components of the homocysteine remethylation pathway and risk of venous thromboembolism. METHODS: We measured concentrations of homocysteine, methionine, and folate in plasma, total folate and methylfolate in red-blood cells, and 5,10 methylenetetrahydrofolate reductase (MTHFR) C677T genotype and other known risk factors for venous thromboembolic disease in 243 patients with deep vein thrombosis or pulmonary embolism and controls matched for sex and age. FINDINGS: Concentrations in plasma of homocysteine differed significantly between cases and controls. We noted a strong concentration-dependent association between concentrations of methylfolate in red-blood cells and risk of venous thromboembolism. The adjusted conditional odds ratio ranged from 1.0 for methylfolate 249 microg/L or greater to 7.1 (3.2-15.8) for methylfolate 141 microg/L or less. Methionine concentrations below the median were also independently associated with raised risk of venous thromboembolic disease, as were established risk factors such as high body-mass index, history of cancer, family history of thromboembolism, oral contraceptive use, and factor V Leiden mutation. Furthermore, the association between concentrations of methylfolate in red-blood cells and risk of thromboembolism varied according to MTHFR C677T genotype. INTERPRETATION: Measurement of methylfolate concentrations in red-blood cells might help to identify people at risk of venous thromboembolism. PMID- 11888586 TI - Association between Staphylococcus aureus strains carrying gene for Panton Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients. AB - BACKGROUND: Between 1986 and 1998, eight cases of community-acquired pneumonia due to Staphylococcus aureus strains carrying the gene for the Panton-Valentine leukocidin (PVL) were recorded in France, six of which were fatal. We aimed to assess the clinical features of these eight cases, and those of other cases identified prospectively, and to compare them with the characteristics of patients with pneumonia caused by PVL-negative strains. METHODS: We compared eight retrospective and eight prospective cases of PVL-positive S aureus pneumonia with 36 cases of PVL-negative S aureus pneumonia. For all patients, we recorded age, length of hospital stay, risk factors for infection, signs and symptoms, laboratory findings, antibiotic treatment, and serial radiological findings. FINDINGS: Median age was 14.8 years (IQR 5.4-24.0) for the PVL-positive patients and 70.1 years (59.2-81.4) for the others (p=0.001). Influenza-like illness had occurred during the 2 days before admission in 12 of the 16 PVL positive patients, but in only three of 33 PVL-negative patients (p<0.001). PVL positive infections were more often marked by: temperature greater than 39 degrees C (p=0.01), heart rate above 140 beats per min (p=0.02), haemoptysis (p=0.005), onset of pleural effusion during hospital stay (p=0.004), and leucopenia (p=0.001). The survival rate 48 h after admission was 63% for the PVL positive patients and 94% for PVL-negative individuals (p=0.007). Histopathological examination of lungs at necropsy from three cases of necrotising pneumonia associated with PVL-positive S aureus showed extensive necrotic ulcerations of the tracheal and bronchial mucosa and massive haemorrhagic necrosis of interalveolar septa. INTERPRETATION: PVL-producing S aureus strains cause rapidly progressive, haemorrhagic, necrotising pneumonia, mainly in otherwise healthy children and young adults. The pneumonia is often preceded by influenza-like symptoms and has a high lethality rate. PMID- 11888587 TI - Fahr's disease. PMID- 11888588 TI - Warts from minor trauma. PMID- 11888589 TI - Localisation of mesenchymal tumours by somatostatin receptor imaging. AB - Oncogenic osteomalacia, an acquired hypophosphataemic syndrome associated with mesenchymal tumours, is characterised by hypophosphataemia secondary to inappropriate phosphaturia, reduced concentrations of serum calcitriol, and defective bone mineralisation. Removal of these tumours results in complete reversal of these biochemical defects. However, because these tumours are small, slow-growing, and frequently situated in unusual anatomical sites, conventional imaging techniques often fail to detect them. Since mesenchymal tumours express somatostatin receptors, we postulated that somatostatin analogues would be able to detect these tumours. We did Indium-111 labeled pentetreotide imaging in seven patients with oncogenic osteomalacia. In five patients, we identified a mesenchymal tumour, and clinical improvement occurred after tumour resection. Our findings suggest that 111In-pentetreotide imaging effectively detects occult mesenchymal tumours and facilitates surgical treatment of oncogenic osteomalacia. PMID- 11888590 TI - Paraoxonase (PON1) polymorphisms in farmers attributing ill health to sheep dip. AB - Human serum paraoxonase (PON1) hydrolyses diazinonoxon, the active metabolite of diazinon, which is an organophosphate used in sheep dip. In a case-referent study, 175 farmers with ill health that they attributed to sheep dip nominated 234 referent farmers who also dipped sheep and whom they believed to be in good health. We calculated odds ratios for polymorphisms in PON1 at positions 192 and 55, and for PON1 activity with diazinonoxon as substrate. Cases were more likely than referents to have at least one R allele at position 192 (glutamine to arginine aminoacid substitution; odds ratio 1.93, 95% CI 1.24-3.01), both alleles of type LL (1.70, 1.07-2.68) at position 55, and to have diazoxonase activity below normal median (1.77, 1.18-2.67). Our results support the hypothesis that organophosphates contribute to the reported ill health of people who dip sheep. PMID- 11888591 TI - Association between Mycoplasma genitalium and acute endometritis. AB - Up to 70% of cases of pelvic inflammatory disease do not have a known cause. We recruited 115 women who had presented to a clinic for sexually transmitted diseases in Nairobi, Kenya with pelvic pain that had persisted for 14 days or less, to look for an association between Mycoplasma genitalium and endometritis. With PCR, we detected M genitalium in the cervix, endometrium, or both in nine (16%) of 58 women with histologically confirmed endometritis and in one (2%) of 57 women without endometritis (p=0.02). Our results suggest that infection with M genitalium is strongly associated with acute endometritis in this population. PMID- 11888592 TI - New HIV drugs show promise in early studies. PMID- 11888596 TI - Microbleeds may predict cerebral bleeding after stroke. PMID- 11888598 TI - African scientists discuss drug-resistant malaria. PMID- 11888599 TI - US health bodies reap funds for bioterrorism. PMID- 11888600 TI - Tobacco company asks Kenyan government to reduce cigarette tax. PMID- 11888601 TI - Spain's regional government becomes first in Europe to sue tobacco industry. PMID- 11888602 TI - CDC chief Koplan quits "the best job in public health". PMID- 11888604 TI - How to limit caesareans on demand--too NICE to push? PMID- 11888605 TI - Evolution of WHO policies for tuberculosis control, 1948-2001. AB - We examine the evolution of WHO managerial policies for tuberculosis control during 1948-2001 to provide a new framework that will accelerate control expansion in the near future. In the first period (1948-63), a vertical approach to tuberculosis control was the policy adopted by WHO and the international community. However, although this approach was successful in more-developed countries, it largely failed in resource-poor settings. As a result, involvement of general health services was soon deemed essential. During 1989-98, a new framework for effective tuberculosis control was created and a new five-element strategy was branded with the name of DOTS. This period was characterised by the recognition of tuberculosis control as a public-health priority, the intensification of tuberculosis control efforts worldwide, and the return of tuberculosis to the political agenda of governments. However, although nominal adoption of DOTS increased rapidly due to massive promotion by WHO and partners, expansion to provide full access was too slow and only 23% of all infectious cases in 1999 were managed under DOTS. A truly multisectoral approach based on advocacy and social mobilisation, community involvement, and engagement of private-for-profit practitioners is becoming the way forward for tuberculosis control. HIV-associated tuberculosis and multidrug-resistant tuberculosis must be tackled as priority issues. We conclude that, based on the lessons of the past, the future of tuberculosis control should be focused on a pragmatic approach combining a specialised, well-defined management system with a fully integrated service delivery. A multisectoral approach that builds on global and national partnerships is the key to future tuberculosis control. PMID- 11888606 TI - Sample size slippages in randomised trials: exclusions and the lost and wayward. AB - Proper randomisation means little if investigators cannot include all randomised participants in the primary analysis. Participants might ignore follow-up, leave town, or take aspartame when instructed to take aspirin. Exclusions before randomisation do not bias the treatment comparison, but they can hurt generalisability. Eligibility criteria for a trial should be clear, specific, and applied before randomisation. Readers should assess whether any of the criteria make the trial sample atypical or unrepresentative of the people in which they are interested. In principle, assessment of exclusions after randomisation is simple: none are allowed. For the primary analysis, all participants enrolled should be included and analysed as part of the original group assigned (an intent to-treat analysis). In reality, however, losses frequently occur. Investigators should, therefore, commit adequate resources to develop and implement procedures to maximise retention of participants. Moreover, researchers should provide clear, explicit information on the progress of all randomised participants through the trial by use of, for instance, a trial profile. Investigators can also do secondary analyses on, for instance, per-protocol or as-treated participants. Such analyses should be described as secondary and non-randomised comparisons. Mishandling of exclusions causes serious methodological difficulties. Unfortunately, some explanations for mishandling exclusions intuitively appeal to readers, disguising the seriousness of the issues. Creative mismanagement of exclusions can undermine trial validity. PMID- 11888607 TI - No problem. PMID- 11888608 TI - Role of HFE in iron metabolism, hereditary haemochromatosis, anaemia of chronic disease, and secondary iron overload. AB - Hereditary haemochromatosis is an iron overloading disorder caused by common mutations in the HFE gene. However, information with respect to the function of HFE protein does not explain how mutations in HFE lead to hereditary haemochromatosis. We propose a molecular model in which HFE has two mutually exclusive activities in cells: inhibition of uptake or inhibition of release of iron. The balance between serum transferrin saturation and serum transferrin receptor concentrations determines which of these functions predominates. With this input, HFE enables the intestinal crypt cells and reticuloendothelial system to interpret the body's iron requirements and regulate iron absorption and distribution. In our model, mutations in HFE result in over absorption of dietary iron, and patterns of tissue iron deposition in agreement with clinical observations of hereditary haemochromatosis. PMID- 11888609 TI - Health-care ethics in Zimbabwe. PMID- 11888610 TI - Detention of asylum seekers in Australia. PMID- 11888611 TI - A body of evidence: torture among asylum seekers to the West. PMID- 11888612 TI - DNA and immigration: the ethical ramifications. PMID- 11888613 TI - Hormone therapy and heart disease. PMID- 11888616 TI - Chinese herbal nephropathy. PMID- 11888617 TI - Embryonic stem cell research in Spain. PMID- 11888618 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888619 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888620 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888621 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888622 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888624 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888625 TI - Mortality in elderly patients with subclinical hyperthyroidism. PMID- 11888626 TI - Target treatment of female urinary dysfunction. PMID- 11888628 TI - Distribution of infectivity in variant Creutzfeldt-Jakob disease. PMID- 11888629 TI - Target treatment of female urinary dysfunction. PMID- 11888630 TI - Complement factor H and haemolytic uraemic syndrome. PMID- 11888631 TI - Treatment of acute migraine. PMID- 11888632 TI - Split testicular function testing in testicular reserve assessment. PMID- 11888633 TI - Fusidic acid resistance in Staphylococcus aureus isolates. PMID- 11888634 TI - Is there a crisis in cardiac transplantation? PMID- 11888635 TI - Hypertension in Japanese old-old. PMID- 11888636 TI - Coxsackie B virus and myocardial infarction. PMID- 11888645 TI - A complement component C3-like protein from the tunicate, Styela plicata. AB - This study identifies a complement component C3-like protein in the solitary tunicate, Styela plicata. Three different polyclonal antibodies raised against C3 molecules from two species (humans and the tunicate, Halocynthia roretzi) were used to identify the C3-like protein in S. plicata hemolymph. The C3 cross reactive protein is a 170kDa heterodimer composed of polypeptides (116 and 80kDa) that have molecular weights comparable to those of C3alpha and C3beta from other species. Amino acid sequencing and amino acid composition analysis confirmed that the C3-like protein from S. plicata is closely related to C3 from H. roretzi. PMID- 11888646 TI - Cell adhesion-related proteins as specific markers of sponge cell types involved in allogeneic recognition. AB - Sponge immunocyte identification is of interest to comparative immunologists since characterizing these cells will allow investigations into the mechanisms of non-self recognition in the oldest animal phylum. Here, we report that polyclonal antibodies raised against the core protein of a proteoglycan involved in cell adhesion in the marine sponge Microciona prolifera are specific markers for archaeocytes, the totipotent sponge cells. Archaeocytes are mobilized upon allogeneic contact and they accumulate in the contact zone. A second type of cell, the gray cells, are specifically recognized by monoclonal antibodies raised against CD44, a hyaluronan receptor. Gray cells do also accumulate in the contact area. Specific staining of a third sponge cell type, the rhabdiferous cells, shows that these do not accumulate upon allografting. These specific cell markers allow tracking of archaeocytes and gray cells, and show that they play an active role in sponge allogeneic reactions. PMID- 11888647 TI - Signaling pathways involved in the physiological response of mussel hemocytes to bacterial challenge: the role of stress-activated p38 MAP kinases. AB - In this work the mechanisms of transduction triggered in Mytilus galloprovincialis hemocytes by bacterial challenge were investigated in an in vitro model of infection of hemocyte monolayers with Escherichia coli. Western blot analyses of hemocyte extracts with phospho-specific anti-MAPK (Mitogen Activated Protein Kinase) antibodies indicate that E. coli induced a time dependent activation of different classes of MAPKs, mainly of the stress activated p38 MAPK. P38 activation was confirmed by the use of the selective kinase inhibitor SB203580. Moreover, hemocyte pretreatment with SB203580 significantly reduced bacterial killing, whereas PD98059, an inhibitor of extracellularly regulated kinase (ERK) activation, was ineffective. Interestingly, the PI3-kinase (phosphatidylinositol-3-OH-kinase) inhibitor, Wortmannin, reduced both p38 activation and bacterial killing, indicating a critical role also for this lipid kinase in the hemocyte immune response. PMID- 11888648 TI - Calreticulin enriched as an early-stage encapsulation protein in wax moth Galleria mellonella larvae. AB - To investigate the molecular mechanism of the early-stage encapsulation reaction in insects, we purified a 47kDa protein from injected beads into Galleria mellonella larvae. When a cDNA clone was isolated, the 47kDa protein showed high homology with Drosophila and human calreticulin. Western blotting analysis showed that the 47kDa protein was present in the hemocytes, but not in the plasma. When the early-stage encapsulated beads were coated with 47kDa protein antibody and reinjected into G. mellonella larvae, any further encapsulation reaction was inhibited. These results suggest that calreticulin is involved in non-self recognition in invertebrate cellular defense reactions. PMID- 11888650 TI - Dynamics of immune cell infiltration in the caecal lamina propria of chickens after neonatal infection with a Salmonella enteritidis strain. AB - Dynamics of leucocyte infiltration and bacterial invasion in the caecal wall were studied after oral infection of 2-day-old chicks with Salmonella enterica ser. Enteritidis. Bacteria invaded the lamina propria of the caecal wall from 12h post challenge onwards. Bacteriological examination of internal organs (liver, spleen) showed a peak in Salmonella bacteria at 3days post-infection, after which the number of bacteria decreased. Immunohistochemistry revealed macrophages and T lymphocytes invading the caecal propria mucosae from 24h after challenge onwards, while B-lymphocytes came somewhat later, subsequently organising into follicular aggregates. An early increase in granulocytes was partly masked by the response to natural flora. While the B-lymphocyte and granulocyte populations were maintained, T-lymphocyte and macrophage populations were already reducing by 10days post-challenge. The infiltration of macrophages and T-lymphocytes in the caecal wall, followed by B-lymphocytes, is the result of an inflammatory response, caused by invading bacteria at this site. The structural maturation of the mucosa-associated lymphoid tissues is antigen driven, since B-cells organise in a follicular pattern. PMID- 11888649 TI - Phagocyte spreading and phagocytosis in the compound ascidian Botryllus schlosseri: evidence for an integrin-like, RGD-dependent recognition mechanism. AB - The involvement of integrins in phagocyte spreading and phagocytosis was investigated in the compound ascidian Botryllus schlosseri. The number of spreading cells was significantly reduced when adhesion occurred in the presence of the tetrapeptide Arg--Gly--Asp--Ser (RGDS), but not of Arg--Gly--Glu--Ser (RGES) indicating the involvement of RGD-mediated adhesion mechanisms in phagocyte spreading. The significant decrease of the fraction of spreading cells in the presence of Botryllus blood plasma suggests the presence of RGD-containing molecules in the blood of our species. The increase in the same index when blood plasma-coated slides as well as fibrinogen- and fibronectin-coated coverslips were used, fits with the above hypothesis. Adhesion in the presence of RGDS leads to a consistent alteration of the actin cytoskeleton, in agreement with the known role of integrin adhesion in microfilament organization. Phagocytosis was greatly reduced by RGDS in the incubation medium, but not by RGES, and was significantly increased by coating yeast cells with fibronectin or blood plasma. Both spreading and phagocytic capability were severely inhibited by wortmannin, suggesting the importance of phosphatidylinositol-3-kinase in integrin-mediated signal transduction in ascidians. PMID- 11888651 TI - Interleukin-7 negatively regulates the development of mature T cells in fetal thymus organ cultures. AB - We added antibody specific for interleukin-7 (IL-7) to chimeric fetal thymus organ cultures (FTOC) to investigate the involvement of this cytokine at distinct stages of T cell development. We report that the neutralization of IL-7 early in fetal T cell development results in a decrease in the production of mature CD4 or CD8 ('single positive', SP) or CD4/8 negative ('double negative', DN) T cell phenotypes, as defined by their expression of CD3. This loss of T cell development was not complete, but it did include the development of gammadelta T cells. However, if IL-7 was neutralized at later stages of FTOC, the production of CD4/8 positive ('double positive', DP) T cells was increased, and if the addition of the antibody was delayed further, the production of mature SP T cells was increased. This last result could be extended to both alphabeta and gammadelta T cells. These data suggested that IL-7 played a negative regulatory role in the development of progressively mature T cells. Tissue sections of FTOC showed that IL-7 was expressed in the subcapsular region of the tissue where immature T cells reside. However, IL-7 was not detected in the medullary region where mature T cells are located. These data suggest that IL-7 not only supports the development of immature fetal T cells, but it may inhibit the development of mature T cells. The production of mature fetal T cells may, therefore, be delayed until their precursors enter the medullary microenvironment, where IL-7 production is low. In this way, T cells may be prevented from maturing until negative selection or anergy events eliminate or inactivate autoreactive clones. PMID- 11888652 TI - Deficiency of antibody responses to T-independent antigens in gerbils---Meriones unguiculatus. AB - Meriones unguiculatus commonly known as gerbils are widely used as animal models for a variety of parasitic infections such as Brugia malayi, Entamoeba histolytica, Giardia duodenalis, Toxoplasma gondi, Helicobacter pylori, Strongyloides stercoralis and Echinococcus multilocularis. Groups of BALB/c mice, gerbils and XID mice were studied for antibody responses to T-independent antigens. Gerbils were found to be significantly deficient in eliciting antibodies to both dextran and phosphorylcholine (PC) in comparison to BALB/c mice. The antibody response of gerbils to T-independent antigens was found to be similar to the response observed in Bruton's tyrosine kinase (Btk) deficient XID mice, which are known to be poor responders to T-independent antigens. Similar to XID mice, normal gerbil sera were found to be deficient in naturally occurring antibodies to single stranded DNA (SS-DNA), lipopolysaccharide (LPS) and phospholipids. This raises the possibility of a deficiency of CD5+ B-lymphocytes (also known as B-1 cells) in gerbils, since deficiency of this sub-population of B-lymphocytes has been attributed to the absence of such naturally occurring antibodies in XID mice. These results indicate the need to study immunogenicity of parasite T-independent antigens and their relationship to protective immunity in parasitic infections in gerbils. PMID- 11888653 TI - Experimental therapy of filovirus infections. PMID- 11888654 TI - Broad-spectrum antiviral activity of PNU-183792, a 4-oxo-dihydroquinoline, against human and animal herpesviruses. AB - We identified a novel class of 4-oxo-dihydroquinolines represented by PNU-183792 which specifically inhibit herpesvirus polymerases. PNU-183792 was highly active against human cytomegalovirus (HCMV, IC(50) value 0.69 microM), varicella zoster virus (VZV, IC(50) value 0.37 microM) and herpes simplex virus (HSV, IC(50) value 0.58 microM) polymerases but was inactive (IC(50) value >40 microM) against human alpha (alpha), gamma (gamma), or delta (delta) polymerases. In vitro antiviral activity against HCMV was determined using cytopathic effect, plaque reduction and virus yield reduction assays (IC(50) ranging from 0.3 to 2.4 microM). PNU 183792 antiviral activity against both VZV (IC(50) value 0.1 microM) and HSV (IC(50) ranging from 3 to 5 microM) was analyzed using plaque reduction assays. PNU-183792 was also active (IC(50) ranging 0.1-0.7 microM) in cell culture assays against simian varicella virus (SVV), murine cytomegalovirus (MCMV) and rat cytomegalovirus (RCMV). Cell culture activity was compared with the appropriate licensed drugs ganciclovir (GCV), cidofovir (CDV) and acyclovir (ACV). PNU-183792 was also active against both GCV-resistant and CDV-resistant HCMV and against ACV resistant HSV. Toxicity assays using four different species of proliferating mammalian cells indicated PNU-183792 was not cytotoxic at relevant drug concentrations (CC(50) value >100 microM). PNU-183792 was inactive against unrelated DNA and RNA viruses indicating specificity for herpesviruses. In animals, PNU-183792 was orally bioavailable and was efficacious in a model of lethal MCMV infection. PMID- 11888655 TI - Antiviral activity of UIC-PI, a novel inhibitor of the human immunodeficiency virus type 1 protease. AB - The human immunodeficiency virus type 1 (HIV-1) protease inhibitor UIC-PI (1) was developed via structure-based design and incorporated a novel bis-tetrahydrofuran (bis-THF) ligand in the (R)-(hydroxyethyl)sulfonamide based isostere. The EC(50) and EC(90) of the compound in acutely-infected H9 cells were <1 and approximately 1 nM, respectively. In chronically infected H9/HIV-1(IIIB) cells, the EC(50) and EC(90) were 20 and 50 nM, respectively. In parallel studies comparing UIC-PI and saquinavir in H9/HIV-1(IIIB) cells, viral p24 levels in culture supernatants were an order of magnitude lower with UIC-PI than with saquinavir. PMID- 11888657 TI - Anti-HIV activity of a glycoprotein from first trimester placental tissue. AB - Human Immunodeficiency Virus-1 (HIV-1) transmission from a mother to the offspring during pregnancy is highly complex and a variable process in part due to the presence of several anti-HIV factors. In this study, we report an early pregnancy-associated protein (early pregnancy associated protein-1, Epap-1) with an apparent molecular weight of 90 kDa in placental tissue of pregnant women during the first trimester. Epap-1 was isolated and purified using Sambucus nigra agarose lectin affinity chromatography. Epap-1 strongly inhibits HIV-1(MN), HIV 1(91US056), HIV-1(VB-7) replication in vitro while it exhibits low activity with HIV-1(VB-66) strain. The molecular analysis of action of Epap-1 shows that it affects HIV-CD4 binding through inhibition of CD4-gp120 interaction. PMID- 11888656 TI - Tenofovir exhibits low cytotoxicity in various human cell types: comparison with other nucleoside reverse transcriptase inhibitors. AB - Clinical studies with tenofovir disoproxil fumarate, an oral prodrug of the nucleotide analog tenofovir, recently approved for the treatment of HIV, have demonstrated antiviral activity and good tolerability in HIV-infected patients. In order to better understand the cytotoxicity profile of tenofovir relative to the other nucleoside reverse transcriptase inhibitors (NRTIs), the in vitro effects of these agents were evaluated in various human cell types. Tenofovir inhibited the proliferation of liver-derived HepG2 cells and normal skeletal muscle cells with CC(50) values of 398 and 870 microM, respectively. In comparison, ZDV, ddC, ddI, d4T, and abacavir all showed lower CC(50) values in these two cell types. Evaluation of hematopoietic toxicity revealed that tenofovir was less cytotoxic towards erythroid progenitor cells (CC(50)>200 microM) than ZDV, d4T, and ddC (CC(50)=0.06-5 microM). Despite some degree of donor-to-donor variability, the inhibitory activity of the tested NRTIs against myeloid cell lineage, in the order of decreasing severity, was consistently ddC>ZDV>d4T>tenofovir>3TC. Finally, tenofovir showed substantially weaker effects on proliferation and viability of renal proximal tubule epithelial cells than cidofovir, a related nucleotide analog with the potential to induce renal tubular dysfunction. In conclusion, tenofovir exhibited weak cytotoxic effects in all cell types tested with less in vitro cytotoxicity than the majority of NRTIs currently used for the treatment of HIV disease. PMID- 11888659 TI - Mathematical modelling in the post-genome era: understanding genome expression and regulation--a system theoretic approach. AB - This paper introduces a mathematical framework for modelling genome expression and regulation. Starting with a philosophical foundation, causation is identified as the principle of explanation of change in the realm of matter. Causation is, therefore, a relationship, not between components, but between changes of states of a system. We subsequently view genome expression (formerly known as 'gene expression') as a dynamic process and model aspects of it as dynamic systems using methodologies developed within the areas of systems and control theory. We begin with the possibly most abstract but general formulation in the setting of category theory. The class of models realised are state-space models, input- output models, autoregressive models or automata. We find that a number of proposed 'gene network' models are, therefore, included in the framework presented here. The conceptual framework that integrates all of these models defines a dynamic system as a family of expression profiles. It becomes apparent that the concept of a 'gene' is less appropriate when considering mathematical models of genome expression and regulation. The main claim of this paper is that we should treat (model) the organisation and regulation of genetic pathways as what they are: dynamic systems. Microarray technology allows us to generate large sets of time series data and is, therefore, discussed with regard to its use in mathematical modelling of gene expression and regulation. PMID- 11888658 TI - Mutations in the UL97 ORF of ganciclovir-resistant clinical cytomegalovirus isolates differentially affect GCV phosphorylation as determined in a recombinant vaccinia virus system. AB - Mutations in the human cytomegalovirus (HCMV) UL97 phosphotransferase have been associated with ganciclovir (GCV) resistance due to an impairment of GCV monophosphorylation. Vaccinia virus recombinants (rVV) were generated that encoded different HCMV UL97 proteins (pUL97) with mutations previously detected in resistant HCMV clinical isolates at codons 460, 520, 592, 594, 595, 598 and 607. These rVVs allowed quantification of GCV phosphorylation catalyzed by the different mutated pUL97s. When compared to rVV-UL97 wild type, mean levels of residual intracellular GCV phosphorylation differed by a factor of 10 for the mutated UL97 proteins ranging from 5.2 to 51.8%. Mutations M460V (located in a UL97 region homologous to domain VIb of protein kinases) and H520Q (located in a cytomegalovirus-specific, functionally critical domain) were responsible for the lowest levels of residual GCV phosphorylation (9.3 and 5.2%). Mutations in a region homologous to the domain IX had a lower impact on GCV phosphorylation (15.8-51.8%). The relevance of pUL97 mutation G598S in inducing GCV resistance was demonstrated for the first time. PMID- 11888660 TI - Many species partial adaptive dynamics. AB - A system of n asexual populations is considered in which both intra- and interspecific frequency-dependent matrix game conflicts with a lack of information are involved and the weak perturbation condition is satisfied (mutation is a very rare event and selection is quick). A new partial adaptive game dynamics is proposed that takes into account that the mutants interact not only with the residents but among themselves as well. It is also shown that if the Nash equilibrium (NE) is totally mixed and the interaction matrices are Replicator-Lyapunov stable (RL-stable), then the NE is an asymptotically stable point of these new dynamics. PMID- 11888661 TI - A predator's selection of an individual prey from a group. AB - This paper examines the mechanisms by which a predator selects an individual target from a group of prey. In a predatory situation, both the group of prey and the predator move around in a two-dimensional space. The predator has to select one individual among these prey to catch. For the mathematical treatment of such a selection, this paper introduces priority functions. Three different priority functions, labeled Strategies N, P and S, are then defined to indicate selection of the Nearest victim, the Peripheral victim or the Split victim (an individual separated from the group), respectively. It is found that, from the predator's standpoint, Strategy P is the best of the three regardless of how the prey group moves in reaction to an attack. Such reaction motions are classified into three types: ordered, partially disordered and fully disordered motion. Of the three types of motion, partially disordered motion is the most difficult type, as it confuses the predator in selecting and tracking a target individual. PMID- 11888662 TI - Quantitative structure-activity relationships by evolved neural networks for the inhibition of dihydrofolate reductase by pyrimidines. AB - Evolutionary computation provides a useful method for training neural networks in the face of multiple local optima. This paper begins with a description of methods for quantitative structure activity relationships (QSAR). An overview of artificial neural networks for pattern recognition problems such as QSAR is presented and extended with the description of how evolutionary computation can be used to evolve neural networks. Experiments are conducted to examine QSAR for the inhibition of dihydrofolate reductase by pyrimidines using evolved neural networks. Results indicate the utility of evolutionary algorithms and neural networks for the predictive task at hand. Furthermore, results that are comparable or perhaps better than those published previously were obtained using only a small fraction of the previously required degrees of freedom. PMID- 11888663 TI - State selection in coupled identical biochemical systems with coexisting stable states. AB - This work examines state selection for coupled biochemical systems with coexisting stable states. For biochemically identical biochemical systems, different coupled systems are examined for the coexistence of (a) one steady state and one oscillatory state or (b) two oscillatory states. For case (a), it is revealed that state selection is always governed by two key factors: the values of kinetic parameters and the coupling strength. When the coupling strength is small, the coupled systems remain in the basin of attraction of their original states. When it is sufficiently large, all coupled systems are always entrained, independently of their original states. Furthermore, for the entrainment, which of the two coexisting states is selected depends sensitively on the activity of recycling enzyme (one of kinetic parameters). It is shown that this is because changing the activity of recycling enzyme alters the size of basin of attraction of each state. When both systems in the same oscillatory state are coupled, an additional factor, namely phase shift between two oscillations, may also affect state selection, and coupling may cause the systems to select either the original oscillatory state or the coexisting steady state. In addition to the features of case (a), case (b) also supports quasiperiodic oscillations and synchronisation of two periodic oscillations. Implications of the results for understanding state selection during the evolution of coupled biochemical systems with coexisting stable states are discussed. PMID- 11888664 TI - Robustness of MetaNet graph models: predicting control of urea production in humans. AB - Urea production in human liver was described by a MetaNet graph, a flowchart-like representation of metabolic pathways that includes parameters for the kinetic constants of the constituent enzymes. Formal operations on the graph facilitate the identification of ligand-binding equilibria that participate in feedback regulation in the network of biochemical reactions. The state of the biochemical network is specified by the concentrations of the intermediates. At any particular time, the influence of an identified locus of regulation is proportional to the respective fractional saturation of the corresponding binding site. Enzymes that make or consume the feedback chemicals share in the control of the strength of the feedback signal in proportion to their fractional saturation. This model predicts control of urea production by the processes that deliver amino groups to the urea cycle enzymes more than by the cycle enzymes themselves. Mitochondrial membrane transport processes are important for transmission of information through the network, but irreversible enzymes and processes far from equilibrium control the strength of the feedback signal. Systematic variation of the parameter values by amounts comparable to the expected variability of their measured values indicated a high probability of invariance in the identities of the predicted control points. The properties of the model are consistent with those of error-tolerant scale-free networks. These results demonstrate the robustness of a MetaNet model's predictions with respect to uncertainties in the values of its parameters. PMID- 11888665 TI - Synergistic tumor promoter effects of estrone and progesterone in methylnitrosourea-induced rat mammary cancer. AB - Tumor promoter effects of steroid hormones were tested in ovariectomized rats treated with the tumor initiator, methylnitrosourea. Continuous treatment with low doses of estrone induced tumor in 55% of experimental animals while intact control animals had a 96-100% incidence. Although progesterone alone produced no tumors in ovariectomized, methylnitrosourea-treated animals, estrone and progesterone acted synergistically, inducing tumorigenesis in 88% of treated rats. These results point to an important interaction between estrogens and progestins in promoting mammary tumorigenesis. PMID- 11888666 TI - Independent variation in susceptibilities of six different mouse strains to induction of pepsinogen-altered pyloric glands and gastric tumor intestinalization by N-methyl-N-nitrosourea. AB - Strain differences in susceptibility regarding stomach carcinogenesis due to N methyl-N-nitrosourea were examined in males of six strains of mice: BALB/cA (BALB), C57BL/6N (C57BL6), CBA/JN (CBA), C3H/HeN (C3H), DBA/2N (DBA/2), and CD-1 (ICR). The frequency of pepsinogen-altered pyloric glands (PAPGs), putative precancerous lesions, was highest (19.6+/-9.9%) in the BALB and lowest in the ICR (12.3+/-5.7%) mice (P<0.05). Incidences of adenocarcinomas at week 52 were 59.3% (16 of 27) and 18.5% (5 of 27), respectively (P<0.005). Invasion also tended to be deepest in BALB compared with the other strains. Intestinal alkaline phosphatase-positive intestinal type cells were observed heterogeneously in some hyperplasias, adenomas and adenocarcinomas consisting of gastric type cells. Thus, intestinalization appeared to occur at random in both non-neoplastic and monoclonal neoplastic lesions, making it unlikely that IAP-positive cells could be precursors of gastric tumors. In contrast, the data suggest a direct histogenetic role for the PAPG, a useful preneoplastic marker lesion in mouse strains. PMID- 11888667 TI - U0126, a mitogen-activated protein kinase kinase inhibitor, inhibits the invasion of human A375 melanoma cells. AB - The anti-invasive ability of the mitogen-activated protein kinase (MAPK) kinase inhibitor, U0126, was examined in human A375 melanoma cells in vitro. The effect was compared to that of PD98059, another commonly used MEK (MAPK kinase) inhibitor. U0126 or PD98059 showed a dose-dependent inhibition of A375 cell invasion through growth factor-reduced Matrigel. U0126 was more potent than PD98059 in suppressing tumor cell invasion. Both compounds significantly decreased urokinase plasminogen activator (uPA) and matrix metalloproteinases-9 (MMP-9) concentrations in conditioned media. At 5 microM, U0126 inhibited phosphorylation of the MEK 1/2 to a non-detectable level within 24 h. The phosphorylation of extracellular signal-related kinase 1/2 was also dramatically suppressed by the treatment with 10 microM U0126 or 40 microM PD98059. Both compounds suppressed the protein expression of c-Jun, but not c-Fos. The expression of uPA and MMP-9 was also inhibited. Our data suggest that U0126 is an effective agent in inhibiting human A375 melanoma cell invasion and that the effect is partially due to the decreased production of uPA and MMP-9. PMID- 11888668 TI - Involvement of the mt1 melatonin receptor in human breast cancer. AB - Two putative melatonin receptors have been described including the cell surface G protein-linked receptors, mt1 and MT2, and the nuclear retinoic orphan receptor alpha (RORalpha). The mt1 receptor, but not the MT2 receptor, is expressed in human breast tumor cell lines, and melatonin-induced growth suppression can be mimicked by the mt1 and MT2 agonist, AMMTC, and blocked by the antagonist, CBPT. RORalpha receptors are also expressed in MCF-7 breast cancer cells and the putative RORalpha agonist CPG-52608 inhibits MCF-7 cell growth but with a very different dose-response than melatonin. Finally, melatonin and AMMTC, but not CPG 52608, can repress RORalpha transcriptional activity in MCF-7 cells. PMID- 11888669 TI - Transferrin overcomes drug resistance to artemisinin in human small-cell lung carcinoma cells. AB - Multiple drug resistance is a significant problem in small-cell lung cancer (SCLC). Artemisinin (ART) is a natural product used to treat drug-resistant malaria. The drug is effective because the Fe2+ present in infected erythrocytes acts non-enzymatically to convert ART to toxic products. We tested the effects of ART on drug-sensitive (H69) and multi-drug-resistant (H69VP) SCLC cells, pretreated with transferrin (TF) to increase the intracellular Fe2+ level. Antibody staining followed by flow cytometry analysis showed twice the level of TF receptors on the H69VP as compared to the H69 cells. Low doses of ART were cytotoxic to SCLC cells. The cytotoxicity of ART for H69VP cells (IC50=24 nM) was ten-fold lower than for H69 cells (IC50=2.3 nM), indicating that ART is part of the drug resistance phenotype. Pretreatment of H69 cells with 220-880 nM TF did not alter the IC50 for ART. However, in the ART-resistant H69VP cells, pretreatment with TF lowered the ART IC50 to near drug-sensitive levels (IC50=5.4 nM after 4 h pretreatment with 880 nM TF). Desferrioxamine (5 microM) inhibited the effect of TF on the IC50 for ART in drug-resistant cells but did not have an effect on ART cytotoxicity in drug-sensitive cells. DNA fragmentation as measured by ELISA occurred within ART-treated cells, with kinetics indicating apoptosis rather than necrosis. This was confirmed by TUNEL staining. These data indicate the potential use of ART and TF in drug-resistant SCLC. PMID- 11888670 TI - Effect of dihydrokainate on the antitumor activity of doxorubicin. AB - For biochemical modulation, components of green tea have been shown to be useful modulators in combination with doxorubicin (DOX). We have confirmed that theanine enhances the antitumor activity of DOX due to inhibition of DOX efflux from tumor cells. Because theanine is a glutamate analogue, we found that it is associated with a change in the drug transport system on the tumor cell membrane, in particular glutamate transporters. We examined the effect of dihydrokainate (DHK), one of the useful glutamate transporter inhibitors. DHK also inhibits DOX efflux significantly and reduces the glutamate uptake by Ehrlich ascites carcinoma cells. The potential contribution of glutamate transporters not only to glutamate uptake but also to cell membrane export of DOX has been shown. In addition, the combination of DHK with DOX significantly enhances the antitumor activity of DOX, by 1.8-fold (P<0.001). The DOX concentration in tumors significantly increases on combination with DHK and is correlated with the reduced tumor weight. On the other hand, DHK tends to reduce the DOX concentration in normal tissues. We expect that DHK has different actions in tumor and normal tissues because different isoforms of glutamate transporters are expressed in the two tissues. Thus, the results suggest that DHK is a novel and useful modulator for inducing enhancement of antitumor activity. PMID- 11888671 TI - Loss of one allele of the p53 gene in the lens epithelial tumor in transgenic mice suppresses apoptosis induced by a topoisomerase I inhibitor (CPT-11). AB - To examine whether CPT-11 can induce apoptosis in the mouse lens tumor, it was administered to pregnant alphaT3 mice, which developed epithelial tumors in the lens during the perinatal stage. Three different p53 genotypes were generated to analyze the influence of p53 status on tumor cells under chemotherapy. On day 16- 17 of gestation, alphaT3 mice received an i.p. injection of CPT-11, and fetal lens tumors were examined 2 days later. Apoptosis in the tumors was observed in both a CPT-11 dose- and p53 gene copy-dependent manner. In addition, it was found that CPT-11 could also induce apoptosis via a p53-independent pathway. PMID- 11888672 TI - p53 codon 72 polymorphism and its association with bladder cancer. AB - p53 codon 72 Arg homozygosity has been associated with increased risk of developing cervical cancer. This association has been tested in various human cancers with controversial results. In the present study we investigated the impact of this polymorphism in a population-based case-control study of bladder cancer. Using allele-specific polymerase chain reaction to detect the p53 codon 72 polymorphism, we tested peripheral blood samples from 50 patients with bladder cancer and 99 healthy individuals of similar age and from the same geographical region. Tumor specimens from all bladder cancer patients were examined for the presence of human papilloma virus (HPV). The distribution of p53 alleles in bladder cancer patients and in controls was statistically significant (P<0.002; odds ratio, 2.67; 95% confidence interval, 1.38-5.20), and homozygosity for arginine at residue 72 was associated with an increased risk for bladder cancer (P<0.00002; odds ratio, 4.69; 95% confidence interval, 2.13-10.41). The presence of HPV was found in six of the 50 patients (12%). This is the first study correlating p53 codon 72 polymorphism with bladder cancer. Our results provide evidence that this p53 polymorphism is implicated in bladder carcinogenesis and that individuals harboring the Arg/Arg genotype have an increased risk of developing bladder cancer. PMID- 11888673 TI - Troglitazone activates p21Cip/WAF1 through the ERK pathway in HCT15 human colorectal cancer cells. AB - In this study, we identified a new mechanism for the anti-proliferation of HCT15 colorectal cancer cells by troglitazone (TRO). Treating HCT15 cells with 20 microM of TRO transiently increased extracellular signal regulated kinase (ERK) activity within 15 min, and this subsequently induced p21Cip/WAF1 cell cycle regulator and localized in the nucleus. Raf-1 modification and MEK activation also occurred after TRO treatment, and Elk-1-dependent trans-reporter gene expression was concomitantly induced. The induction and nuclear localization of p21Cip/WAF1 by TRO were blocked by PD98059 pre-treatment, which suggested a role for the ERK pathway in p21Cip/WAF1 activation. TRO inhibited BrdU incorporation and no BrdU incorporation was observed in most p21Cip/WAF1-activated cells. Therefore, TRO regulates the proliferation of HCT15 cells at least partly by a mechanism involving the activation of p21Cip/WAF1. PMID- 11888674 TI - Serum VEGF levels in patients undergoing primary radiotherapy for cervical cancer: impact on progression-free survival. AB - Vascular endothelial growth factor (VEGF) plays an important role in the regulation of tumour growth and metastasis. It was the aim of this study to examine the impact of serum VEGF levels on the likelihood of response to radiotherapy and on the disease-free survival in patients with cervical cancer. Blood was taken before commencing treatment and serum VEGF was assessed by quantitative ELISA in 23 patients with cervical cancer stage IB-IVA undergoing primary radiotherapy. Serum VEGF levels were correlated with clinical and histopathologic factors as well as with response to radiotherapy and time to progression. Nineteen of the 23 patients had a complete response and four patients had persistent disease at 3 months. The median follow-up was 25 months (95% confidence interval: 23.5-26.5 months). At the time of analysis, eight patients were tumour-free and 15 patients had tumour progression; 12 of these 15 patients died of disease. Overall, the median serum VEGF level was 244 pg/ml (range 31.9-817.6 pg/ml). All four patients with local failure had VEGF levels >244 pg/ml, whereas 11 of the 19 patients with complete response had serum VEGF of < or =244 pg/ml (P=0.035). The median time to progression was 5 months in patients with VEGF of >244 pg/ml compared to 19 months in patients with VEGF of < or = 244 pg/ml (log rank, P=0.003). In multivariate analysis, serum VEGF, tumour size and histological type, but not the patient's age, stage and grade of histological differentiation influenced the progression-free survival. Elevated pre-therapeutic serum VEGF levels are associated with poor response and a shorter time to progression in patients with cervical cancer undergoing primary radiotherapy. PMID- 11888675 TI - Abrogation of IRF-1 response by high-risk HPV E7 protein in vivo. AB - We have previously reported that human papillomavirus (HPV) E7 interacts with IRF 1, a key regulator of cellular immune response, and abrogates its transactivation function at the molecular level in vitro. To confirm our previous data, we investigated in vivo the E7-mediated down-regulation of IRF-1 using HPV E7 inducible cells and transgenic mice expressing HPV-18 E6/E7. When E7 was induced in the absence of tetracycline, the expression of target genes of IRF-1 (TAP-1, IFN-beta, MCP-1 that are important for cellular immunity) was clearly reduced as determined by RT-PCR. In addition, the IRF-1 activity was down-regulated in E7 expressing cells into which IFN-beta-CAT reporter plasmid was transfected. To further investigate the E7-mediated immune regulation in vivo, we constructed transgenic mice expressing E6 and E7 genes of HPV-18 under the control of HPV-18 promoter (URR). From several rounds of breeding, we obtained from a transgenic line that developed a cervical dysplasia and expressed E6/E7 as determined by histological examination and RT-PCR, respectively. Subsequent RT-PCR analysis indicated that TAP-1, IFN-beta, and MCP-1 genes were less expressed in a cervical tissue derived from transgenic mouse, when compared with a cervix derived from normal mouse. From these results, we conclude that the E7 transgene expression inactivates the transactivation function of IRF-1 in vivo, which might be important for the elucidation of the E7-mediated immune evading mechanism that is frequently found in cervical cancer. PMID- 11888676 TI - Angiogenesis in health and disease. PMID- 11888677 TI - The discovery of angiogenic factors: a historical review. AB - Angiogenesis is a biological process by which new capillaries are formed and it occurs in many physiological and pathological conditions. It is controlled by the net balance between molecules that have positive and negative regulatory activity and this concept had led to the notion of the "angiogenic switch," depending on an increased production of one or more of the positive regulators of angiogenesis. Numerous inducers of angiogenesis have been identified and this review offers a historical account of the relevant literature concerning the discovery of the best-characterized angiogenic factors. PMID- 11888678 TI - Experimental models of growth factor-mediated angiogenesis and blood-retinal barrier breakdown. AB - Following chronic ischemia, vascular endothelial growth factor (VEGF) is induced primarily in the ganglion cell layer of the retina. This often results in neovascularization (NV) that originates from the vascular bed closest to the ganglion cell layer. To study the effects of VEGF, independent lines of transgenic mice that express VEGF in the lens and in the retina have been generated. Expression in the lens results in excessive proliferation and accumulation of angioblasts and endothelial cells in proximity to the lens. However, VEGF expression is not sufficient to direct blood vessel organization or maturation in the prenatal mouse. Abnormal vessels do form on the retinal surface, but not until the second postnatal week. In transgenic mice expressing VEGF in the photoreceptors, NV originates from the deep capillary bed--the vascular bed closest to the photoreceptors. NV is accompanied by localized blood retinal barrier breakdown. NV is also induced in PDGF-B transgenic mice. PDGF-B expression in the lens occurs prenatally and, during this time, mainly affects the perilenticular vessels. Postnatally, transgenic mice expressing PDGF-B in the lens or photoreceptors show a similar phenotype. In both models, a highly vascularized cell mass containing endothelial cells, pericytes, and glia forms in the superficial retina, and the formation of the deep capillary bed is inhibited. The phenotype suggests that an additional factor is necessary for the maturation and penetration of vascular endothelial cells into the retina to form the deep capillary bed. PMID- 11888679 TI - Effects of thrombin and of the phospholipase C inhibitor, D609, on the vascularity of the chick chorioallantoic membrane. AB - Microvascular corrosion casting was used to assess the effects of thrombin and D609, a phospholipase C inhibitor, on the vascularity of the chick embryo chorioallantoic membrane (CAM). Discs containing vehicle, thrombin or D609 were placed on the CAM of fertilized white Leghorn eggs on Day 9 of gestation and vascularity was assessed on Day 11. Thrombin caused significant increases in the numbers (43%), diameters (5%) and lengths (17%), of both pre- and postcapillaries (first-order vessels by centripetal ordering). Conversely, D609 caused a decrease in the numbers (27%), lengths (12%) and diameters (8%) of first-order vessels. D609 decreased the total vascular volume of first- to third-order vessels by 32%, whereas thrombin increased vascular volume by 27%. Additionally, thrombin increased capillary plexus density by 6%, whereas D609 decreased capillary plexus density by 3%. These findings provide a quantitative assessment of changing vascularity in the chick CAM--a model assay system in the development of pro- and antiangiogenic agents. PMID- 11888680 TI - Thrombin peptide, TP508, stimulates angiogenic responses in animal models of dermal wound healing, in chick chorioallantoic membranes, and in cultured human aortic and microvascular endothelial cells. AB - The alpha-thrombin peptide, TP508, accelerates the healing of full-thickness wounds in both normal and ischemic skin. In wounds treated with TP508, a pattern of increased vascularization is consistently observed both grossly and microscopically when compared to wounds treated with saline. One possible mechanism by which the peptide accelerates wound healing is by promoting revascularization of granulation tissue at the injured site. To evaluate the angiogenic potential of TP508, the peptide was tested in the chick embryo chorioallantoic membrane (CAM), where it increased the density and size of CAM blood vessels relative to controls. Additionally, TP508 stimulated chemokinesis and chemotaxis in a dose-dependent fashion in cultured human aortic and human microvascular endothelial cells. Taken together, these in vivo and in vitro data support an angiogenic role for TP508 in wound healing. A working model is presented to explain how this 23-amino-acid peptide, which lacks proteolytic activity, is generated during wound healing and contributes to the nonproteolytic functions associated with alpha-thrombin during tissue repair. PMID- 11888681 TI - Thrombin regulates the expression of proangiogenic cytokines via proteolytic activation of protease-activated receptor-1. AB - In addition to its central role in blood coagulation and hemostasis, human alpha thrombin is a growth factor for a variety of cell types, including monocytes and endothelial cells, involved in the control of angiogenesis. Different cytokines produced by mononuclear cells have been implicated in angiogenic processes associated with tissue repair and certain human malignancies. We have previously shown that thrombin enhances proliferative responses in T lymphocytes. More recently, we reported that interferon-gamma-differentiated monocytes have increased expression of protease-activated receptor-1 (PAR-1) and increased thrombin binding. Since cytokines may be involved directly and indirectly in angiogenesis, we initiated studies to determine thrombin effects on the induction of cytokines, such as interleukin (IL)-1 and IL-6, in human mononuclear cells. IL 1 and IL-6 protein expression was significantly enhanced by thrombin (P<.05), as determined by enzyme-linked immunosorbent assay (ELISA). Treating mononuclear cells with the PAR-1 peptide, SFLLRN, has effects similar to those of thrombin. Thus, it appears that these thrombin effects are mediated through activation of PAR-1. These results confirm that thrombin is a strong activator of monocytes and could be involved in angiogenesis by inducing cytokines that could enhance the angiogenic process in tissue repair. PMID- 11888682 TI - Locating the active site for angiogenesis and cell proliferation due to fibrin fragment E with a phage epitope display library. AB - The plasmin-mediated lysis of fibrin present in a wound, or in chronic inflammatory disease, results in the release of fibrin degradation products. One of the two major products is fibrin fragment E, which has been shown to stimulate cell proliferation in many cell types including endothelium, fibroblasts, and smooth muscle cells, and to be angiogenic in the chick chorioallantoic membrane (CAM) system. The activity of fibrin fragment E is dependent on N-terminus thrombin action. Antibodies against fibrin E, which block the cell proliferative activity, were used to locate the active site. Phage epitope display libraries were used to identify the sequence of a peptide, which resembles a region of the N terminus structure. The equivalent synthetic peptide (WTM110) has optimal stimulatory properties at equimolar concentrations to the parent molecule. Such peptide information has therapeutic potential for both stimulating and suppressing angiogenesis and cell proliferation. PMID- 11888683 TI - The role of peroxisome proliferator-activated receptor gamma in bladder cancer in relation to angiogenesis and progression. AB - Peroxisome proliferator-activated receptor gamma (PPAR gamma) immunohistochemical expression was analyzed in 75 human bladder tumor specimens, where the expression of some angiogenic factors, such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived endothelial cell growth factor (PDECGF), and tumor progression markers, such as epidermal growth factor receptor (EGFr), p16, mutated p53, and normal pRB, were also analyzed. The results were then compared to the clinical and pathological characteristics of the disease. PPAR gamma was expressed more significantly in papillary tumors than in solid cancers, and its presence was associated with statistical significance to low incidence of tumor recurrence or progression. This significant association was observed also when PPAR gamma was expressed in the presence of PDECGF, which resulted, when considered alone, to an angiogenic factor typical of solid cancers and appeared related to poor prognosis. In the presence of bFGF, on the contrary, PPAR gamma expression no longer resulted to a significant association with low incidence of tumor recurrence or progression, suggesting a possible worsening role of this angiogenic factor, typical of papillary cancers, in its interaction with PPAR gamma. PMID- 11888684 TI - Effects of microenvironmental extracellular pH and extracellular matrix proteins on angiostatin's activity and on intracellular pH. AB - Antiangiogenic agents target migratory and proliferative endothelial cells (EC) in the process of forming new vessels, resulting in growth inhibition or cell death. Here we have shown that the antiangiogenic activity of angiostatin on EC is enhanced in culture when the microenvironmental extracellular pH (pH(e)) is reduced to levels similar to that of many tumors. In a migration/scratch assay and during tube formation, angiostatin in combination with reduced pH(e) synergistically resulted in an increased EC death--an effect not seen with either stimulus individually. Lowering of pH(e) decreased intracellular pH (pH(i)), and a further lowering of pH(i) occurred when low pH(e) was combined with angiostatin. These data suggest that low pH(e) plays a role in the relative specificity and efficacy of angiostatin for tumor neovasculature and indicate roles for both pH(e) and pH(i) in the mechanism of angiostatin action. A receptor for angiostatin, the alpha-subunit of ATP synthase, was found on the surface of EC. We show that cell surface receptor distribution is increased on Matrigel, a basement-like matrix, as opposed to fibronectin or RGD peptide substrates, and redistributed to a more punctuate appearance at low pH(e). Furthermore, positive cell surface histochemical staining for alpha-ATP synthase was blocked by preincubation with angiostatin. These data indicate that substrate and pH(e) are critical parameters in the evaluation of this antiangiogenic substance, and probably for others as well. PMID- 11888685 TI - Directional spread of an alpha-herpesvirus in the nervous system. AB - Pseudorabies virus (PRV), an alpha-herpesvirus, is capable of spreading between synaptically connected neurons in diverse hosts. In this report, two lines of experimentation are summarized that provide insight into the mechanism of virus spread in neurons. First, techniques were developed to measure the transport dynamics of capsids in infected neurons. Individual viral capsids labeled with green fluorescent protein (GFP) were visualized and tracked as they moved in axons away from infected neuronal cell bodies in culture during egress. Second, the effects of three viral membrane proteins (gE, gI and Us9) on the localization of envelope, tegument, and capsid proteins in infected, cultured sympathetic neurons were determined. These three proteins are necessary for spread of infection from pre-synaptic neurons to post-synaptic neurons in vivo (anterograde spread). Us9 mutants apparently are defective in anterograde spread in neural circuits because essential viral membrane proteins such as gB are not transported to axon terminals to facilitate spread to the connected neuron. By contrast, gE and gI mutants manifest their phenotype because these proteins most likely function at the axon terminal of the infected neuron to promote spread. These two sets of experiments are consistent with a model for herpesvirus spread in neurons first suggested by Cunningham and colleagues where capsids and envelope proteins, but not whole virions, are transported separately into the axon. PMID- 11888686 TI - Mechanisms of pathogenesis in herpetic immunoinflammatory ocular lesions. AB - This article reviews possible mechanisms by which ocular infections with herpes simplex virus result in a blinding immunoinflammatory lesion in the cornea. We conclude that this immunoinflammatory response involves multiple immune mechanisms including autoimmunity. PMID- 11888687 TI - Towards non-invasive imaging of HSV-1 vector-mediated gene expression by positron emission tomography. AB - The overall goals of the broad and growing field of molecular medicine is to identify fundamental errors of disease and to develop corrections of them on the molecular level. At the same time, real-time imaging of gene expression in vivo aims towards a detailed analysis of both endogenous and exogenous gene expression in animal models of disease and in the clinical setting. Non-invasive imaging of endogenous gene expression may reveal insight into the molecular basis of disease pathogenesis and the extent of treatment response. When exogenous genes are introduced, e.g. by herpes simplex virus type 1 (HSV-1)-based vectors, to ameliorate a genetic defect or to add an additional gene function to cells, imaging techniques may reveal the assessment of the location, magnitude and duration of therapeutic gene expression and its correlation to the therapeutic effect. Here, we review the main approaches of non-invasive imaging techniques of gene expression in vivo with special reference to HSV-1 vector-mediated gene expression. PMID- 11888688 TI - Glycoprotein M of bovine herpesvirus 1 (BHV-1) is nonessential for replication in cell culture and is involved in inhibition of bovine respiratory syncytial virus F protein induced syncytium formation in recombinant BHV-1 infected cells. AB - Cell cultures infected with BHV-1/F(syn), a recombinant bovine herpesvirus 1 (BHV 1) which expresses a synthetic open reading frame encoding the fusion (F) protein of the bovine respiratory syncytial virus (BRSV), showed a cytopathic effect (CPE) indistinguishable from that induced by wildtype BHV-1 although transient transfection experiments demonstrated that expression of the F protein leads to formation of large syncytia. Since it has been shown that glycoprotein M (gM) of pseudorabies virus inhibits BRSV F-induced syncytium formation in transient plasmid transfection experiments [Pseudorbies virus glycoprotein M inhibits membrane fusion. J. Virol. 74 (2000) 6760], the gM ORF of wtBHV-1 and BHV 1/F(syn) was interrupted. Infection of cell cultures with the resulting gM(-) mutant of BHV-1/F(syn) led to formation of syncytia, whereas the CPE in gM(-)BHV 1 infected cells was comparable to the CPE in wtBHV-1 infected cultures. Our results demonstrate that gM is not essential for BHV-1 replication in cell culture and that gM is involved in inhibition of the cell fusion activity of the BHV-1 expressed BRSV F protein. PMID- 11888689 TI - Antibody-induced internalization of viral glycoproteins in pseudorabies virus infected monocytes and role of the cytoskeleton: a confocal study. AB - Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins. PMID- 11888691 TI - Evolution of the herpesviruses. AB - The complete genome sequences of 26 herpesvirus species, some represented by more than one strain, are currently deposited in the public databases. Their combined length totals over four million base pairs. Several additional genomes are in the pipeline, and a host of partial sequence information is also available. Consequently, researchers have a very detailed picture of the genetic content of herpesviruses and their relationships to each other. This review of the herpesvirus phylogenetic tree explains what is currently known about the evolution of this important virus family by proceeding from the twigs, along the branches, boughs and trunk and towards the root. The account focuses naturally on places where the grasp is secure, but also ventures where the bark is slippery and looks out on regions where footholds have not yet been established. PMID- 11888690 TI - Improvement of serological discrimination between herpesvirus-infected animals and animals vaccinated with marker vaccines. AB - Control/eradication plans of bovine herpesvirus 1 (BHV1) and suid herpesvirus 1 (SHV1) infections involve vaccination with inactivated or attenuated gE-deleted marker vaccines and associated companion serological tests to discriminate naturally infected from vaccinated animals. Blocking or competitive enzyme-linked immunosorbent assays (ELISAs) have been designed for the detection of specific antibodies against BHV1 or SHV1 gE glycoprotein. The antigen source usually consists of a crude viral preparation in which gE is associated with other envelope glycoproteins. Such assays suffer from a lack of specificity which is not due to serological cross-reactions with other pathogens. Interestingly, false positive results occur with sera collected from multivaccinated cattle or pigs. After multivaccination with a marker vaccine, the binding of the conjugated monoclonal antibody used as a tracer, could be hampered by antibodies directed against the other viral glycoproteins. In order to validate the steric hindrance hypothesis, a simple preadsorption of such samples was carried out with a preparation of antigen devoid of gE, prior to the blocking ELISA itself. The decrease in antibody concentrations against the major glycoproteins, clearly leads to a better discrimination between positive and negative samples; that is between infected and multivaccinated animals, without significant loss of sensitivity. This experiment confirms the steric hindrance hypothesis, therefore serum preadsorption could be an easy way to improve the specificity of currently available diagnostic tests. PMID- 11888692 TI - Temporary disturbance of actin stress fibers in swine kidney cells during pseudorabies virus infection. AB - Rounding and loosening of cells is a consequence of infection with pseudorabies virus (PrV), both in vitro and in vivo. These changes in the normal structure of the cell may be the result of cytoskeletal changes. Immunofluorescence staining of actin filaments and microtubule bundles was performed to examine whether PrV induces a reorganization of these cytoskeletal components in infected swine kidney (SK) cells. Every 2h until 12h post-inoculation (p.i.), cells were washed in cytoskeleton stabilizing buffer (CSB), fixed with paraformaldehyde and washed again with CSB. Cells were permeabilized with a 1/1000 dilution of Triton X-100 and actin filaments were stained by incubating cells with phalloidin-Texas Red. Staining of microtubules was done by incubating the cells subsequently with mouse monoclonal anti-alpha-tubulin and goat anti-mouse IgG-FITC. During the course of infection, actin fibers of SK cells were rearranged in the following sequence: (1) disappearance of thick actin stress fibers between 4 and 6h p.i., (2) complete loss of stress fibers between 6 and 8h p.i., and (3) reappearance of thin stress fibers starting from 10h p.i. In contrast to herpes simplex virus 1 (HSV1) or equine herpesvirus 1 (EHV1), PrV infection did not induce changes in the cellular microtubule network. PrV infection induces a temporary disassembly of actin stress fibers. PMID- 11888693 TI - Search for physical interaction between BICP0 of bovine herpesvirus-1 and p53 tumor suppressor protein. AB - The immediate-early (IE) protein BICP0 of bovine herpesvirus-1 (BHV-1) may have other functions besides transactivation of viral promoters. Recently, we observed that BICP0, delivered to cultured cells by a helpervirus-free amplicon system, forms spherical or doughnut-like structures in which the tumor suppressor protein p53 is sequestered. The objective was to determine whether BICP0 and p53 interact physically, we used both yeast and mammalian two-hybrid systems. As a bait plasmid, pVA3 which encodes a hybrid protein consisting of the Gal4 DNA binding domain fused to murine p53 was used. The BICP0 gene or its truncated versions were inserted into the prey plasmid pGAD424. Bait and prey plasmids were cotransformed into yeast strain Y153, which has LacZ and HIS3 reporter genes under the control of Gal4 upstream activating sequence. After 4-6 days, colonies were stained for beta-galactosidase activity. In the mammalian two-hybrid system, pM-53 was used as a bait where truncated p53 fused to Gal4 DNA binding domain is expressed. The BICP0 gene was cloned into prey plasmid pVP16. The interaction between p53 and SV40 T-antigen was evaluated as a positive control in both systems. Neither full-length BICP0 nor its truncated derivatives induced beta galactosidase activity in yeast whereas the positive control turned blue under the same conditions. The mammalian two-hybrid system, in which chloramphenicol acetyltransferase (CAT) activity was used as a reporter, also failed to show an interaction between these two proteins. Co-localization of p53 with BICP0 in spherical structures is unlikely to result from a direct physical interaction between these two proteins. Mediation by additional cellular proteins may be required. PMID- 11888694 TI - HSV-1-based amplicon particles are able to transduce cells of feline origin with genes encoding biologically functional feline IL-10 or IL-6. AB - The most common viral disease of cats worldwide is the infection with feline herpesvirus 1 (FeHV-1). This infection may be followed by Herpetic stromal keratitis (HSK), which is supposed to have an immunopathological basis. Experiments using herpes simplex viruses (HSV) in mouse models indicated that HSK may be treated by topical application of the interleukin 10 (IL-10) gene. The objective of this study was the construction of human herpes simplex virus type 1 (HSV-1)-based amplicon vectors expressing feline interleukin genes and delivery of these genes into cells of feline origin. HSV-1-based amplicon vectors encoding either the enhanced green fluorescent protein, the feline IL-6 or the feline IL 10 under control of the HSV-1 immediate-early 4/5 promotor were constructed, packaged into amplicon particles, transduced into feline cells, and tested for RNA synthesis and biological activity. Feline cells were successfully transduced by HSV-1-based amplicon particles and RNA specific for the transgene was detected already at 2h post transduction, with a maximum at 24h. The recombinant feline IL 10 was functionally active as demonstrated by the reduction of both IL-12 p40 and interferon-gamma-mRNA production in Pansorbin stimulated feline peripheral mononuclear cells. Similarly, the recombinant feline IL-6, which was secreted into the supernatant of transduced cells, was able to support the growth of the IL-6-dependent murine B cell hybridoma 7TD1. HSV-1-based amplicon particles are able to transduce cells of feline origin with genes encoding biologically functional feline IL-10 or IL-6. It will be of high interest to study the effects of these tools in vivo. PMID- 11888695 TI - Simultaneous intramammary and intranasal inoculation of lactating cows with bovine herpesvirus 4 induce subclinical mastitis. AB - In this study, we examined whether an experimental bovine herpesvirus 4 (BHV4) infection can induce bovine mastitis, or can enhance bovine mastitis induced by Streptococcus uberis (S. uberis). Four lactating cows were inoculated intramammarily and intranasally with BHV4, and four lactating control cows were mock-inoculated. After 14 days, two of four cows from each group were inoculated intramammarily with S. uberis. No clinical signs were recorded in cows inoculated only with BHV4, and their milk samples showed no abnormal morphology, despite the fact that BHV4 replicated in inoculated quarters. Somatic cell count increased significantly in milk from three of six BHV4-inoculated quarters, compared to the non-inoculated quarters of the same cows (within-cow) and the quarters of mock inoculated cows (control group) on days 8, 9 and 11 post-inoculation (pi). BHV4 was isolated from nasal swabs between days 2 and 9 pi. Clinical mastitis was observed in all four cows intramammarily inoculated with S. uberis. A preceding BHV4 infection did not exacerbate the clinical mastitis induced by S. uberis. S. uberis infections appeared to trigger BHV4 replication. From one quarter of each of two cows inoculated with BHV4 and S. uberis, BHV4 was isolated, and not from quarters inoculated with BHV4 only. In conclusion, BHV4 did not induce bovine clinical mastitis after simultaneous intranasal and intramammary inoculation. However, the BHV4 infection did induce subclinical mastitis in 50% of the cows and the quarters. PMID- 11888696 TI - Otarine herpesvirus-1: a novel gammaherpesvirus associated with urogenital carcinoma in California sea lions (Zalophus californianus). AB - The incidence of neoplasia in California sea lions (CSLs) is considered to be unusually high. Electron microscopic examination of some of these urogenital tumours revealed the presence of virions with typical herpes-like structure. While current attempts to cultivate this virus have not been successful, molecular studies employing DNA extracted from tumour tissues allowed both the classification of the agent and its identification in tumours and archived tissue samples. Two genome fragments generated using degenerate primers in PCR demonstrated highest identities with other mammalian gammaherpesviruses. Phylogenetic analysis showed that this novel virus, tentatively designated Otarine herpesvirus-1 (OtHV-1), grouped with members of the gammaherpesvirus subfamily and was distinct from PHV-2, a previously described pinniped gammaherpesvirus. An OtHV-1 specific PCR was established and used to investigate the presence of this virus in CSL tissues. PCR of DNA isolated from animals with these tumours, demonstrated that this virus was present in 100% (16/16) of tumours. Furthermore, DNA extracted from archived brain and muscle tissues was also positive in 29% (4/14) and 50% (7/14) of cases examined. This preliminary study provides evidence to support the hypothesis that the presence of this novel gammaherpesvirus is a factor in the development of urogenital carcinoma in CSLs. PMID- 11888697 TI - Analysis of bovine trigeminal ganglia following infection with bovine herpesvirus 1. AB - Following primary infection of the eye, oral cavity, and/or nasal cavity, bovine herpesvirus 1 (BHV-1) establishes latency in trigeminal ganglionic (TG) neurons. Virus reactivation and spread to other susceptible animals occur after natural or corticosteroid-induced stress. Infection of calves with BHV-1 leads to infiltration of lymphocytes in TG and expression of IFN-gamma (interferon-gamma), even in latently infected calves. During latency, virus antigen and nucleic acid positive non-neural cells were occasionally detected in TG suggesting there is a low level of spontaneous reactivation. Since we could not detect virus in ocular or nasal swabs, these rare cells do not support high levels of productive infection and virus release or they do not support virus production at all. Dexamethasone (DEX) was used to initiate reactivation in latently infected calves. Foci of mononuclear or satellite cells undergoing apoptosis were detected 6h after DEX treatment, as judged by the appearance of TUNEL+ cells (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling). BHV-1 antigen expression was initially detected in lymphocytes and other non-neural cells in latently infected calves following DEX treatment. At 24h after DEX treatment, viral antigen expression and nucleic acid were readily detected in neurons. Our data suggest that persistent lymphocyte infiltration and cytokine expression occur during latency because a low number of cells in TG express BHV-1 proteins. Induction of apoptosis and changes in cytokine expression following DEX treatment correlates with reactivation from latency. We hypothesize that inflammatory infiltration of lymphoid cells in TG plays a role in regulating latency. PMID- 11888698 TI - Increased susceptibility of peripheral blood mononuclear cells to equine herpes virus type 1 infection upon mitogen stimulation: a role of the cell cycle and of cell-to-cell transmission of the virus. AB - Equine herpesvirus-1 (EHV-1) is an important pathogen of horses, causing abortion and nervous system disorders, even in vaccinated animals. During the cell associated viremia, EHV-1 is carried by peripheral blood mononuclear cells (PBMC), mainly lymphocytes. In vitro, monocytes are the most important fraction of PBMC in which EHV-1 replicates, however, mitogen stimulation prior to EHV-1 infection increases the percentage of infected lymphocytes. The role of the cell cycle in viral replication and the role of cluster formation in cell-to-cell transmission of the virus were examined in mitogen-stimulated PBMC. Involvement of the cell cycle was examined by stimulating PBMC with ionomycin/phorbol dibutyrate (IONO/PDB) during 0, 12, 24 and 36 h prior to inoculation. Cell cycle distribution at the moment of inoculation and the percentage of EHV-1 antigen positive PBMC at 0, 12 and 24 hours post inoculation (hpi) were determined by flow cytometry and immunofluorescence microscopy, respectively. The role of clusters was examined by immunofluorescence staining within clusters of stimulated PBMC using antibodies against EHV-1. Significant correlations were found between the increase of cells in the S- or G2/M-phase after a certain time interval of prestimulation and the increase of EHV-1 antigen-positive cells. The percentage of clusters with adjacent infected cells significantly increased from 3.3% at 8 hpi to 23.7% at 24 hpi and the maximal number of adjacent infected cells increased from 2 to 7. Addition of anti-EHV-1 hyperimmune serum did not significantly alter these percentages. Mitogen stimulation favours EHV-1 infection in PBMC by: (i) initiating cell proliferation and (ii) inducing formation of clusters, thereby facilitating direct cell-associated transmission of virus. PMID- 11888699 TI - Transduction of Vero cells and bovine monocytes with a herpes simplex virus-1 based amplicon carrying the gene for the bovine herpesvirus-1 Circ protein. AB - Herpes simplex virus-1 (HSV-1) based amplicon vectors are promising gene delivery vehicles because they have a large transgene capacity and can efficiently transduce many different cell types, including non-dividing cells, of various animal species. The Circ protein of bovine herpesvirus-1 (BHV-1) is a myristylated virion component of unknown function. Preliminary experiments with a circ gene deletion mutant indicated that Circ may influence the host's immune response by downregulating MHC-II expression in bovine monocytes. To get more insight into the function of Circ, amplicon vectors were constructed with various open reading frames (ORFs) under the control of the HSV-1 IE4/5 promoter: (i) the Circ ORF alone, (ii) a fusion ORF encoding an N-terminal Circ fused to the enhanced green fluorescent protein (eGFP), (iii) the eGFP ORF alone, and (iv) the Circ ORF in the inverted orientation. Upon helpervirus-free packaging into HSV-1 amplicon particles and transduction of Vero cells, both Circ alone and the Circ eGFP fusion protein produced a punctate pattern within the cytoplasm, suggesting membrane association of the myristylated protein. In contrast, eGFP alone was evenly distributed over the cytoplasm of transduced cells. Upon infection of bovine buffy-coat cells, it was observed that cells of the monocyte lineage but not lymphocytes were transduced. Transgene expression reached a peak around 20h after transduction and lasted for at least 90h. Transduced monocytes underwent specific morphological changes, which may be attributed to Circ synthesis. PMID- 11888700 TI - Herpesviruses: balance in power and powers imbalanced. AB - The first veterinary herpesvirus symposium, organized under the patronage of the European Society for Veterinary Virology (ESVV) and the Swiss Societies for Microbiology (SGM-SSM), was held at the University of Zurich, Switzerland, on 22nd and 23rd of March 2001. The congress was divided into six sessions. The first session was dedicated to introductory lectures towards the main topics of the symposium, namely pathogenesis, immune response, and gene therapy. Session 2 was committed to new insights into herpesvirus-related gene therapy and vaccination. Specific and general aspects of the immune response against herpesviruses were presented in session 3, while session 4 was dedicated to virus replication. Session 5 was dedicated to a variety of poster presentations. Finally, session 6 revealed new insight into the pathogenesis of different herpesviruses. The present article summarizes the contributions and draws a new view of the herpesviruses. The herpesviruses have apparently found a multi dimensionally balanced position between the powers of "cytopathogenicity" and "tumorigenicity" on one hand and "immunogenicity" and "tolerogenicity" on the other hand. As long as the different powers stay in balance, no or little clinical disease may be expected in association with herpesvirus infections. However, unbalanced actions of those powers may lead to disease. PMID- 11888701 TI - Effect of fumonisin B1 on structure and function of macrophage plasma membrane. AB - Fumonisin B1 (FB1), a mycotoxin produced by Fusarium moniliforme and related fungi, is nephrotoxic, neurotoxic, hepatotoxic, carcinogenic and immunosuppressive in animals and man. In this study we evaluate the modifications of fluidity, endocytosis and peroxidative damage of plasma membrane induced by FB1 in macrophage cell line J774A.1. In these immune cells FB1 (1-10 microM) enhances membrane fluidity and increases, time-dependently, the horseradish peroxidase (HRP) endocytosis. This effect is concentration-dependent, significant at 10 microM, and reverted by IFN-gamma (100 U/ml). Moreover, FB1 (1-10 microM) induces a membrane peroxidative damage as evident by the increase of malondialdehyde (MDA) production. All these mycotoxin effects provide additional insight into potential mechanism by which FB1, in macrophages, might enhance membrane damage and oxidative stress contributing to the pathogenesis of mycotoxin induced diseases. PMID- 11888702 TI - Maternal-fetal distribution of cadmium in the guinea pig following a low dose inhalation exposure. AB - Pregnant guinea pigs in their last trimester of gestation were exposed by inhalation to cadmium (Cd) chloride level (50 microg/m3 Cd) for 1 and 5 days. Cd content was evaluated by atomic absorption spectrophotometry. Maternal blood Cd concentration increased by 127 and 223% of control for 1 and 5 days of exposure. Maternal lung Cd concentration increased significantly by 11.66- and by 48.24 fold after 1 and 5 days of treatment, while maternal liver showed an increase of 188 and 227% for 1 and 5 days of exposure. Also, fetal Cd concentration significantly increased in brain (156 and 192%), liver (159 and 174%) and heart (201 and 157%) after 1 and 5 days of exposure, compared to unexposed females. Placental calcium content decreased significantly by 16% of control after 5 days of exposure. These results suggest that low-level inhalation of Cd may pass through the guinea pig placenta and accumulate in fetal brain, liver and heart. PMID- 11888703 TI - Detection of phytoestrogens in samples of second trimester human amniotic fluid. AB - There is widespread concern that fetal exposure to hormonally active chemicals may adversely affect development of the reproductive tract. Therefore, the present study was performed to develop the necessary analytical methods and test the hypothesis that dietary phytoestrogens can be quantified in second trimester human amniotic fluid. Amniotic fluid samples (n=59) from women (n=53) undergoing routine amniocentesis between 15 and 23 weeks of gestation were analyzed by gas chromatography/mass spectrometric (GC/MS). Analytes included the phytoestrogens daidzein, genistein, formononetin, biochanin A, and coumestrol. Dietary phytoestrogens were quantified in 96.2% of second trimester amniotic fluid samples tested. The mean (+/- standard deviation (S.D.)) concentration of daidzein and genistein in amniotic fluid was 1.44 +/- 1.34 and 1.69 +/- 1.48 ng/ml with maximum levels of 5.52 and 6.54 ng/ml, respectively. Second trimester amniotic fluid contains quantifiable levels of dietary phytoestrogens and thus is a marker of mid pregnancy fetal exposure. PMID- 11888704 TI - Production of gamma-hydroxybutyric acid in postmortem liver increases with time after death. AB - Gamma-hydroxybutyric (GHB) acid, which is becoming popular as a drug of abuse, was shown by means of gas chromatography-mass spectrometry (GC-MS) to increase in mouse liver with time after death. The amount detected was 0.8 +/- 1.0 microg/g at 3 h after death, 4.7 +/- 1.5 microg/g at 24 h, and 8.8 +/- 0.8 microg/g at 72 h. Furthermore, GHB was detected in samples from deceased persons, at concentrations of 2.6-12.0 microg/g in liver, 0.4-7.3 microg/ml in blood, and 0 2.6 microg/ml in urine, but was not detected in the blood and urine of living persons. Although 1,4-butanediol has been suggested to be a precursor of GHB produced after death, 1,4-butanediol was not detected in any of our samples. Additionally, succinate semialdehyde arising from gamma-aminobutyric acid (GABA) transamination to GHB was also barely detectable in any of our samples. This study supports previous reports that GHB is a product of postmortem decomposition. Production of GHB increases with time after death in postmortem liver; since we were unable to identify endogenous 1,4-butanediol and succinate semialdehyde in our samples, the pathway of GHB production after death remains unclear. PMID- 11888705 TI - Influence of oxygenated fuel additives and their metabolites on the binding of a convulsant ligand of the gamma-aminobutyric acid(A) (GABA(A)) receptor in rat brain membrane preparations. AB - As a foundation for evaluating potential mechanisms of the neurological effects (e.g. headache, nausea, dizziness) of some octane boosters, we studied the gamma aminobutyric acid(A) (GABA(A)) receptor in a series of binding assays in membranes from rat brain. The GABA(A) receptor was probed using the radioligand [3H]t-butylbicycloorthobenzoate ([3H]TBOB) which binds to the convulsant recognition site of the receptor. The results demonstrated that the short-chain t ethers and their t-alcohol metabolites inhibit binding at the convulsant site of the GABA(A) receptor. The potency of the inhibition tended to correlate with carbon chain length. For agents having an equal number of carbon atoms, potency of inhibition of [3H]TBOB binding was greater in magnitude for the alcohols than for the ethers. The descending rank order of potency for the ethers and alcohols were as follows, t-amyl alcohol (TAA); t-amyl-methyl ether (TAME); ethyl-t-butyl ether (ETBE)>t-butyl alcohol (TBA)>methyl-t-butyl ether (MTBE)>ethanol. In additional saturation binding assays, MTBE reduced apparent density of convulsant binding (B(max)). PMID- 11888706 TI - Rat liver microsomal and nuclear activation of methanol to hydroxymethyl free radicals. AB - Recent studies from other laboratories reported that during methanol intoxication lipid peroxidation and protein oxidation in liver occurred. Further, they detected free radicals-PBN adducts in bile and urine of methanol poisoned rats. In this work, we report the presence in liver microsomes and nuclei of NADPH dependent processes of hydroxymethyl (HMet) radical formation. The detection of HMet radicals was performed by GC/MS of the trimethylsilyl derivatives of the PBN (N-tert-butyl-a-phenylnitrone)-radical adducts. The formation of HMet radicals was observed only under nitrogen, in these in vitro conditions. Formation of formaldehyde from methanol was observed in aerobic incubation mixtures containing either microsomes or nuclei but also under nitrogen using microsomes. The latter process was not inhibited by diphenyleneiodonium while the anaerobic microsomal one producing HMet was strongly inhibited by it. This shows that they are independent processes. Results suggest that both, liver nuclei and microsomes are able to generate free radicals during NADPH-mediated methanol biotransformation. PMID- 11888707 TI - The association between frequencies of mitomycin C-induced sister chromatid exchange and cancer risk in arseniasis. AB - In order to examine whether biomarkers of cytogenetic damage and susceptibility, such as spontaneous and mitomycin C-induced sister chromatid exchange (SCE) can predict cancer development, a nested case-control study was performed in a blackfoot endemic area with known high cancer risk. A cohort of 686 residents was recruited from three villages in the arseniasis area. Personal characteristics were collected and venous blood was drawn for lymphocyte culture and stored in a refrigerator. The vital status and cancer development was followed using the National Death Registry, Cancer Registry, and Blackfoot Disease Registry. The follow up period was from August 1991 to July 1997. During this 6-year-period, 55 residents developed various types of cancer. Blood culture samples from 23 of these subjects were unsuitable for spontaneous SCE experiments and 45 of these subjects were unsuitable for mitomycin C-induced SCE experiments due to improper storage. Finally, a total of 32 cancer cases had cytogenetic samples that could be analyzed. About 32 control subjects were selected from those who did not develop cancer in the study period and these subjects were matched to cases by sex, age, smoking habits, and residential area. The results showed that there was no significant difference in the frequencies of spontaneous and mitomycin C induced SCE between the case and control groups. There was also no significant difference in the net difference of spontaneous and mitomycin C-induced SCE between the case and control groups. These results suggest that SCEs, either spontaneous or mitomycin C-induced, might not be good markers to predict cancer risk. PMID- 11888708 TI - Acute cytotoxicity of the chemical carcinogen 2-acetylaminofluorene in cultured rat liver epithelial cells. AB - Acute cytotoxic effects of 2-acetylaminofluorene (AAF), a powerful chemical carcinogen, were studied in cultured rat liver epithelial F258 cells. Acute treatment of these cells by AAF resulted in inhibition of cell proliferation, through an arrest in G0/G1 phase of the cell cycle, and a loss of cell viability. By contrast, AAF failed to trigger apoptosis as demonstrated by flow cytometric analysis of hypoploid sub-G1 cells. Cytochrome P4501A1 (CYP1A1), a drug metabolizing enzyme thought to play a major role in biotransformation of AAF, was induced in AAF-treated F258 cells as assessed by Northern blotting. AAF cytotoxic effects were however not blocked by the CYP1A1 inhibitor alpha-naphtoflavone, thus suggesting that they did not require CYP1A1 activity. They were also not prevented by the antioxidant N-acetylcysteine, making unlikely a major contribution of AAF-related reactive oxygen species. These data therefore indicate that AAF can exert acute cellular toxicity, including inhibition of cell growth and cell death, in rat liver epithelial cells without triggering an apoptotic process. Such an acute toxicity may contribute to the well-known promoting effects of AAF. PMID- 11888709 TI - Genetic susceptibility of term pregnant women to oxidative damage. AB - Genetic polymorphisms involved in the activation and detoxification of exogenous chemicals and in the production and scavenging of reactive oxygen species may modulate the levels of oxidative injury biomarker. We investigated 81 pregnant women in Inchon, Korea. In addition to a questionnaire survey, urinary concentrations of 8-hydroxydeoxyguanosine (8-OH-dG) and malondialdehyde (MDA) were measured as oxidative injury biomarkers. Cytochrome P-450(CYP)1A1, CYP2E1, glutathione S-transferase (GST)M1 and GSTT1 polymorphisms and myeloperoxidase (MPO) and manganese superoxide dismutase (MnSOD) polymorphisms were evaluated to determine the effect of genetic modification on urinary 8-OH-dG and MDA. The concentrations of urinary 8-OH-dG were significantly elevated in the presence of the MnSOD variant genotype (P=0.04) and in the case of GSTM1 null status (P=0.02) by multivariate regression. The concentrations of urinary MDA were not affected significantly by the genetic polymorphisms. This result shows that oxidative stress injury is modified by some heritable polymorphisms, including GSTM1 and MnSOD. PMID- 11888710 TI - Accumulation of methylsulfonylmethane in the human brain: identification by multinuclear magnetic resonance spectroscopy. PMID- 11888712 TI - Re: video-assisted thoracoscopic sympathectomy-splanchnicectomy. PMID- 11888714 TI - Methadone for refractory cancer pain. PMID- 11888715 TI - Cryoneuroablation for pain in a 12-year-old girl. PMID- 11888716 TI - Descriptors of breathlessness in patients with cancer and other cardiorespiratory diseases. AB - The objective of this study was to examine the relationship between descriptors of breathlessness and its underlying cause in patients with lung cancer and cardiopulmonary diseases to see whether descriptors might be used to help determine the cause of breathlessness, particularly in patients with lung cancer. We studied 131 patients with primary or secondary lung cancer, whose breathlessness was attributed to tumor mass, pleural effusion, lung collapse, metastases, pleural thickening or lymphangitis carcinomatosis, and 130 patients with breathlessness attributed to asthma, chronic obstructive pulmonary disease (COPD), interstitial lung disease or cardiac failure. Patients selected statements (descriptors) that described the quality of their breathlessness from a 15-item questionnaire and the relationship between the descriptors and the attributed cause of breathlessness was evaluated by cluster analysis. All patient groups were characterized by more than one cluster and several clusters were shared between groups. Specific sets of clusters were associated with breathlessness due to asthma, COPD and cardiac failure, and to cancer causing collapse, metastases or pleural thickening. The association of different sets of clusters with the different diagnostic groups suggests that patients are describing qualitatively different experiences of breathlessness, but the relationship does not appear to be sufficiently robust for the questionnaire to aid differential diagnosis. PMID- 11888717 TI - Dynamic cancer pain management outcomes: the relationship between pain severity, pain relief, functional interference, satisfaction and global quality of life over time. AB - To examine the relationship between different cancer pain management outcomes over time, 74 patients with the worst cancer related pain rated as four or greater on an 11-point numeric scale were followed weekly with the Brief Pain Inventory (BPI), and the satisfaction questionnaire and global visual analogue scale quality of life (VASQOL) for 3 weeks. Univariate and multivariate regression analyses were performed at weekly time points. The analyses indicated that pain outcomes can be categorized into separate QOL and satisfaction paths linked by the worst pain severity. In the QOL path, the worst pain severity predicted a pain interference score, which consistently predicted VASQOL. For the satisfaction path, independent predictors were pain relief at Week 1, and worst pain severity and changes in worst pain severity at Week 2. No variables predicted satisfaction at Week 3. The data suggest that satisfaction and quality of life may be independent outcomes of pain management. The timing of assessment may itself be important. PMID- 11888718 TI - Measuring the quality of children's postoperative pain management: initial validation of the child/parent Total Quality Pain Management (TQPM) instruments. AB - Quality improvement measurement instruments for pediatric postoperative pain management are virtually nonexistent. Without standardized instruments to measure pediatric pain management outcomes, practitioners are hampered in their efforts to improve the quality of pain management for children. In this study, instruments for children (8--12 years) and parents were developed and tested to measure the quality of children's postoperative pain management. The child (Child TQPM) and parent (Parent TQPM) Total Quality Pain Management instruments were tested with 50 parent/child dyads across two large treatment centers. The pain rating scale modified for these instruments demonstrated good criterion validity with the well established Varni/Thompson Pediatric Pain Questionnaire Visual Analogue Scale. Parent--child agreement was described for responses across instruments. Construct validity was examined through selected inter-item relationships. Psychometric analyses support the initial measurement properties of the pediatric TQPM instruments. PMID- 11888719 TI - Probing the paradox of patients' satisfaction with inadequate pain management. AB - The paradox of patients who are in pain, yet satisfied with their pain management, has been previously reported. To probe this paradox, we used cross sectional data collected in the primary care setting on cancer patients' patterns of pain and pain treatment, beliefs and expectations about pain and pain relief, willingness to report pain and take pain medication, care from the provider, and satisfaction with their pain management (n = 316). Descriptive findings were similar to other studies: more than 75% of patients were satisfied or very satisfied with their overall pain management, despite almost half of all patients reporting recent moderate to severe pain. Univariate and bivariate analyses were consistent with the hypothesis that patients may expect and are therefore satisfied with the "peak and trough" pattern of pain severity that occurs with "as-needed" administration of analgesics. However, multivariate analyses failed to directly support this hypothesis. Instead, regression analyses identified factors related to characteristics of patients' pain experiences, patients' beliefs about pain and its inevitability, the frequency that patients reported their pain, and aspects of the patient--provider relationship. Predictors of patients' satisfaction with how their primary care doctor managed their pain included: whether or not the patient was told that treating pain was an important goal, whether or not the patient reported sustained long-term pain relief, and the degree to which the patient was willing to take opioids if prescribed by the doctor or nurse (adjusted R(2) = 0.22). Qualitative data collected from patients who were in severe pain during the past three days but satisfied with their pain management (n = 88) further suggest the importance of the patient--provider relationship in shaping patient expectations. Based on these findings, we recommend that future research on outcomes in pain management place greater emphasis on the potential impact of the patient-provider relationship. PMID- 11888720 TI - Prevalence of pain in Italian hospitals: results of a regional cross-sectional survey. AB - The aims of this study were to quantify the prevalence of pain among hospitalized Italian patients and to describe the potential determinants of pain in this population. All patients older than 18 years and hospitalized for at least 24 hours in one of the 30 public hospitals of the Liguria region (n = 4709) were eligible for pain assessment. Using the Brief Pain Inventory, patients with pain during the last 24 hours were asked to score the intensity of pain at the time of the interview, and the worst pain and average pain during the previous 24 hours on 0-10 rating scales. Overall, 87% (4121 / 4709) of inpatients were interviewed, and 56.6% suffered pain during the last 24 hours. Among patients with pain, the median (interquartile range) score of the worst and of the average pain during the last 24 hours was 7 (5-9) and 5 (3-6), respectively. At the time of interview, 43.1% evaluated patients suffered pain, with a median (interquartile range) of 5 (3-7). Although significant heterogeneity in the distribution of pain was observed among the hospitals, pain prevalence was unacceptably high in most cases. Age, sex, education, diagnosis, and days from surgery were significantly related to pain prevalence in univariate analyses. In a multivariable ordinal regression logistic analysis, only sex, diagnosis, days from surgery, and hospitals remain significantly associated with increased pain prevalence. PMID- 11888721 TI - Pain management and prescription monitoring. AB - Preventing diversion and abuse of prescription controlled substances while ensuring their availability for legitimate medical use is an important public health goal in the United States. In one approach to preventing and identifying drug diversion, 17 states have implemented prescription monitoring programs (PMPs) to monitor the prescribing of certain controlled substances. While PMPs are not intended to interfere with legitimate prescribing, some in the pain management community feel that they negatively affect prescribing for pain management. This article describes a collaborative project initiated by the Pain & Policy Studies Group that brought together regulatory and pain management representatives twice in 1998 to share perspectives and reconcile differing views on the effects of PMPs. The ultimate goals of this project are to provide accurate information to healthcare clinicians about PMPs, better define the balance between preventing drug diversion and providing pain management, and promote continued dialog and cooperation among the groups. PMID- 11888722 TI - Pain measurement tools and methods in clinical research in palliative care: recommendations of an Expert Working Group of the European Association of Palliative Care. AB - An Expert Working Group was convened under the auspices of the Steering Committee of the Research Network of the European Association of Palliative Care to review the status of the use of pain measurement tools (PMTs) in palliative care research conducted in a multilingual-multicenter setting. Based on a literature review and on the experts' opinion, the present work recommends that standardized methods should be applied for the use of PMTs in research in palliative care. Visual analogue scales, numerical rating scales, and verbal rating scales are considered valid to assess pain intensity in clinical trials and in other types of studies. Among the multidimensional questionnaires designed to assess pain, the McGill Pain Questionnaire and Brief Pain Inventory are valid in many multilingual versions. Specific recommendations for PMT use and administration, depending on the study type and aim, are reviewed. Special population requirements specific of clinical situations encountered in palliative care (elderly, terminal, cognitively impaired patients, pediatric patients) are also considered. PMID- 11888724 TI - Application of conducting polymers to biosensors. AB - Recently, conducting polymers have attracted much interest in the development of biosensors. The electrically conducting polymers are known to possess numerous features, which allow them to act as excellent materials for immobilization of biomolecules and rapid electron transfer for the fabrication of efficient biosensors. In the present review an attempt has been made to describe the salient features of conducting polymers and their wide applications in health care, food industries, environmental monitoring etc. PMID- 11888723 TI - When midazolam fails. AB - Significant distress is experienced by patients, families, and caregivers when a symptom or disorder, such as an agitated delirium, becomes an intractable, or a catastrophic event, such as irreversible stridor. When palliative sedation is indicated for these patients, midazolam is usually the preferred drug. In some cases, however, midazolam fails to provide adequate sedation. Two cases are presented to illustrate this phenomenon and explore the possible mechanisms underlying this lack of response. These mechanisms appear to be multifaceted. The heterogeneity of the GABA(A) receptor complex and the alterations that this complex can undergo functionally can explain, to some degree, the diversity of the physiological and pharmacological outcomes. Other factors responsible for the diversity in response may include concomitant medications, age, concurrent disease, overall health status, alcohol use, liver disease, renal disease, smoking and hormonal status. Evidence-based guidelines on alternative treatment options should midazolam fail are required. In the interim, a lower threshold for adding an alternative drug, such as phenobarbital, or substituting midazolam with another drug, such as propofol, should be considered in these circumstances. PMID- 11888725 TI - Reagent-less detection of a competitive inhibitor of immobilized acetylcholinesterase. AB - The interaction of monosulfonate tetraphenyl porphine (TPPS(1)) with immobilized acetylcholinesterase (AChE) yields a characteristic absorbance peak at 446 nm. Addition of acetylcholine iodide or the competitive inhibitor tetracaine to the immobilized TPPS(1)-AChE complex results in a decrease in absorbance intensity at 446 nm due to displacement of the porphyrin from the active site. The loss in intensity at 446 nm is linearly dependent on tetracaine concentration at levels below 100 ppb. Tetracaine concentrations as low as 300 ppt have been detected. PMID- 11888726 TI - Flow system for fish freshness determination based on double multi-enzyme reactor electrodes. AB - A double reactor system for the determination of fish and shellfish freshness using the freshness indicator, K-value (K=[(HxR+Hx)/(ATP+ADP+AMP+IMP+HxR+Hx)]x100), was developed, where ATP, ADP, AMP, IMP, HxR and Hx are adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine monophosphate, inosine and hypoxanthine, respectively. The system consisted of a pair of enzyme reactors with an oxygen electrode positioned close to the respective reactor. The enzyme reactor (I) was packed with nucleoside phosphorylase and xanthine oxidase immobilized simultaneously on chitosan beads (immobilized enzyme A). Similarly, the enzyme reactor (II) was packed with immobilized enzyme A and immobilized enzyme B (co-immobilized alkaline phosphatase and adenosine deaminase). Moreover, this reactor consisted of two layers, the enzyme A and enzyme B (1:1). A good correlation was obtained between K values, which were determination by the proposed system and by the HPLC method. One assay could be completed within 5 min. The signal for the determination of K value of fish and shellfish was reproducible within 2.3%. The long-term stability of the enzyme reactors was evaluated at 30 degrees C for 28 days. PMID- 11888728 TI - Measurement of blood viscosity using mass-detecting sensor. AB - A newly designed mass-detecting capillary viscometer is extended to measure the viscosity of whole blood over a range of shear rates without the use of anticoagulants in a clinical setting. In the present study as proof of principle, a single measurement of liquid-mass variation with time replaces the flow rate and pressure drop measurements that are usually required for the operation of a capillary tube viscometer. Using a load cell and capillary, we measured the change of mass flowing through capillary tube with respect to the time, m(t), from which viscosity and shear rate were mathematically calculated. For water and adulterated bloods, excellent agreement was found between the results from the mass-detecting capillary viscometer and those from a commercially available rotating viscometer. Also, the mass-detecting capillary viscometer measured the viscosity of unadulterated whole blood without heparin or EDTA. This new method overcomes the drawbacks of conventional viscometers in the measurement of the whole blood viscosity. First, the mass-detecting capillary viscometer can accurately and consistently measure the unadulterated blood viscosity over a range of shear rates in less than 2 min without any anticoagulants. Second, this design provides simplicity (i.e. ease of operation, no moving parts, and disposable) and low cost. PMID- 11888727 TI - Sterilization of enzyme glucose sensors: problems and concepts. AB - A useful method of enzyme glucose sensor sterilization has not only to ensure the needs of sterility assurance but has also to guarantee the functional stability of the sensors. The action of 2 or 3% alkalinized glutaraldehyde solution, as well as gamma irradiation with a dose of 25 kGy caused changes of the in vitro functionality and polymer material irritations, respectively. After a combined treatment by 0.6% hydrogen peroxide solution acting over 4 days with 7 kGy gamma irradiation only a slight loss of sensitivity must be registered. The combination of a specially designed universal homogeneous ultraviolet irradiation over 300 s with a 3 days lasting treatment by an inclusion compound of hydrogen peroxide with tensides in urea (0.15% effective hydrogen peroxide concentration) did not cause any influence on the glucose sensor function in vitro. With all methods tested here, a Bacillus subtilis spore reduction over 8 log(10) cycles from 10(6) to 10(-2) spores per test object on an average could be proved experimentally. In general, if non-thermal methods must be used it seems to be impossible to guarantee a sterility assurance level of 10(-6) as it is demanded by the pharmacopoeias. Consequently, effective concepts to produce sterile glucose biosensors for medical and biological applications should be based not only on final product treatments but should include germ reducing measures in every manufacturing step. PMID- 11888729 TI - Development of disposable amperometric sulfur dioxide biosensors based on screen printed electrodes. AB - The possibility of developing amperometric biosensors for the measurement of SO(2) in flowing gas streams has been examined. Screen-printed carbon electrodes (SPCEs) were tailored with the enzyme sulfite oxidase and cytochrome c and the response is generated through the resulting enzymatic and electrocatalytic reactions involving SO(3)(2-), formed when SO(2) gas is dissolved in the supporting electrolyte. Two methods of integrating the enzyme and cytochrome c with the SPCE were investigated. In one design (b-type biosensor), the components were mixed thoroughly with the same ink used to produce the SPCEs, then the modified ink was spread over the working electrode. In the second approach the bio-components were dissolved in the supporting electrolyte and simply deposited on top of the transducer (s-type biosensor). Both devices gave linear responses over the range 4--50 ppm but the sensitivity of the s-type was approximately twice that of the b-type biosensor. In addition, the time taken to reach 90% of the maximum response (t(90%)) was 110 s for the s-type biosensor compared with 200 s for the b-type biosensor. These studies illustrate the successful use of biosensors for the detection of sulfur dioxide at the relatively low potential of +0.3 V versus Ag.AgCl and should provide useful alternatives for decentralised environmental studies. PMID- 11888730 TI - Gas sensor arrays for early detection of infection in mammalian cell culture. AB - The detection of bacterial infections in a mammalian cell culture process is realised using a gas sensor array. In production-scale and laboratory-scale cultivations of a perfused recombinant CHO-cell culture producing human blood coagulation Factor VIII, we show that the gas sensor array identifies bacterial contamination earlier than conventional methods. The sensitivity of the instrument is verified by inoculation of a blank cell culture medium with defined bacterial cell counts. PMID- 11888731 TI - Immobilization of homooligonucleotide probe layers onto Si/SiO(2) substrates: characterization by electrochemical impedance measurements and radiolabelling. AB - Radiolabelling and electrochemical impedance measurements were used to characterize the immobilization of single stranded homooligonucleotides onto silica surfaces and their subsequent hybridization with complementary strands. The immobilization procedure consists of grafting an epoxysilane onto microelectronic grade Si/SiO(2) substrates, and coupling oligonucleotides bearing a hexylamine linker onto the epoxy moiety. Radiolabelling was used as a reference method to quantify the amount of immobilized and hybridized oligonucleotides. These results show that the Si/SiO(2) substrates modified with an epoxysilane yield a surface concentration of approximately 10(11) strands/cm(2) for the immobilized oligonucleotides, after vigorous washings, and that approximately 36% of these undergo hybridization with complementary strands. The impedance measurements, which provide a direct means of detecting variations in electrical charge accumulation across the semiconductor/oxide/electrolyte structure when the oxide surface is chemically modified, show that the semiconductor's flat band potential undergoes reproducible shifts of -150 and -100 mV following the immobilization and the hybridization step, respectively. These results demonstrate that electrochemical impedance measurements using chemically modified semiconductor/oxide/electrolyte structures of this type offer a viable alternative for the direct detection of complementary DNA strands upon hybridization. PMID- 11888732 TI - Laser microfabricated model surfaces for controlled cell growth. AB - The relatively recent applications of microelectronics technology into the biological sciences arena has drastically revolutionized the field. New foreseeable applications include miniaturized, multiparametric biosensors for high performance multianalyte assays or DNA sequencing, biocomputers, and substrates for controlled cell growth (i.e. tissue engineering). The objectives of this work were to investigate a new method combining microphotolithographical techniques with laser excimer beam technology to create surfaces with well defined 3-D microdomains in order to delineate critical microscopic surface features governing material-cell interaction. Another obvious application of this study pertains to the fabrication of cell-based biosensors. Microfabricated surfaces were obtained with micron resolution, by "microsculpturing" polymer model surfaces using a laser excimer KrF beam coupled with a microlithographic projection technique. The laser beam after exiting a mask was focused onto the polymer target surface via an optical setup allowing for a 10-fold reduction of the mask pattern. Various 3-D micropatterned features were obtained at the micron level. Reproducible submicron features could also be obtained using this method. Subsequently, model osteoblast-like cells were plated onto the laser microfabricated surfaces in order to study the effects of particular surface microtopography on preferential cell deposition and orientation. Preferential cell deposition was observed on surfaces presenting "smooth" microtopographical transitions. This system may provide an interesting model for further insights into correlations between 3-D surface microtopography and cell response with new applications in the field biosensor, biomaterial and pharmaceutical engineering sciences (e.g. new cell based biosensors, controlled synthesis of immobilized cell derived active ingredients). PMID- 11888733 TI - Enhancement in the sensitivity of a gas biosensor by using an advanced immobilization of a recombinant bioluminescent bacterium. AB - A genetically engineered bioluminescent bacterium (lac::luxCDABE) was immobilized to develop a whole cell biosensor for the detection of toxic gaseous chemicals. The toxicity of chemicals can be evaluated through the bioluminescent reaction as it reduces in intensity when the cells experience toxic or lethal conditions. This whole cell biosensor was fabricated, using an immobilization technique utilizing solid agar medium, for the measurement of toxicity through direct contact of the cells with the gas. To enhance the sensitivity of the biosenor, glass beads were used and the thickness of the agar layer was reduced. The bioluminescent response was measured using a fiber optic probe connected between the biosensor kit and a luminometer. As sample gaseous toxic chemicals, BTEX (Benzene, Toluene, Ethylbenzene, and Xylene) gases were selected and their vapors were produced by a gas generation system. The concentrations of the gaseous chemicals injected into the chamber were controlled by the time of exposure and were measured using a portable gas chromatograph (Allstech., USA). Additions of glass beads facilitated gas diffusion through the solid medium, making the biosensor more sensitive. In addition, a thinner matrix layer was more advantageous for the detection of gas toxicity. PMID- 11888734 TI - A portable toxicity biosensor using freeze-dried recombinant bioluminescent bacteria. AB - A portable biosensor has been developed to meet the demands of field toxicity analysis. This biosensor consists of three parts, a freeze-dried biosensing strain within a vial, a small light-proof test chamber, and an optic-fiber connected between the sample chamber and a luminometer. Various genetically engineered bioluminescent bacteria were freeze-dried to measure different types of toxicity based upon their modes of action. GC2 (lac::luxCDABE), a constitutively bioluminescent strain, was used to monitor the general toxicity of samples through a decrease in its bioluminescence, while specific toxicity was detected through the use of strains such as DPD2540 (fabA::luxCDABE), TV1061 (grpE::luxCDABE), DPD2794 (recA::luxCDABE), and DPD2511 (katG::luxCDABE). These inducible strains show an increase in bioluminescence under specific stressful conditions, i.e. membrane-, protein-, DNA-, and oxidative-stress, respectively. The toxicity of a sample could be detected by measuring the bioluminescence 30 min after addition to the freeze-dried strains. In an attempt to enhance the sensitivity of the freeze-dried cells, glucose and Tween 80 were tested as additives. It was found that the addition of glucose had a negative effect on the viability of the freeze-dried cells, while samples having Tween 80 showed an increase in their viability. On the other hand, the addition of either Tween 80 or glucose decreased the final bioluminescent response of DPD2540 in response to 4-chlorophenol. Using these strains, many different chemicals were tested and characterized. This portable biosensor, with a very simple protocol, can be used for field sample analysis and the monitoring of various water systems on-site. PMID- 11888735 TI - Air travel related venous thromboembolism--an existing problem that can be prevented? PMID- 11888736 TI - The economic evaluation of arterial intervention. PMID- 11888737 TI - Primary closure of median sternotomy: techniques and principles. AB - Understanding the biomechanics of midline sternotomy repair is important to successful surgical outcome. High-risk patients, particularly those with immunosuppression, diabetes, and osteoporosis, should be identified. Details of technique should be monitored, and closure should incorporate lateral support of the sternum in patients at risk. Sternal dehiscence can occur under physiologic loads. Closure techniques and materials should insure stable repair, with avoidance of material migration through the bone. PMID- 11888738 TI - Combined coronary artery bypass and carotid endarterectomy: long-term results. AB - PURPOSE: We determined late survival, freedom from late stroke, and freedom from late cardiac events in patients treated by combined coronary artery bypass and carotid endarterectomy (CAB/CEA). METHODS: All patients who underwent CAB/CEA in our institution between January 1994 and December 1999 were identified. Follow-up data were obtained from office records and telephone interviews. Endpoints included death from any cause, stroke, and non-fatal cardiac events (MI, CHF, percutaneous transluminal angioplasty with stenting, redo CAB). Data were expressed in life table format. RESULTS: Over a 6-yr period 154 patients had combined CAB/CEA with a 3.9% postoperative stroke rate. Six patients (3.9%) died, leaving 148 patients for follow-up. Average follow-up was 38 +/- 23 months (range: 1-82 months). During the follow-up period two patients (1.4%) had late strokes and 17 patients (11%) had late non-fatal cardiac events. The late mortality rate was 13% (19 patients). Of the late mortalities, four were related to cardiac disease and one to stroke. Using Kaplan-Meier analysis, the 5-yr survival probability was 80 +/- 4.3%. The freedom from late ipsilateral neurologic events was 98 +/- 1.3% at 5 yr. The freedom from late cardiac events was 82 +/- 4.6% at 5 yr. CONCLUSIONS: The large majority of patients with combined coronary and carotid artery disease can be expected to live for greater than 5 yr. Therefore, these patients should be considered candidates for prophylactic CEA for stroke prevention, even when their carotid lesions are asymptomatic. Successful CAB/CEA provides good long-term survival and freedom from late cardiac events, as well as excellent freedom from late stroke. Further reduction in perioperative events will make this operative approach even more attractive in patients with combined disease. PMID- 11888739 TI - The role of intraoperative digital subtraction angiography for quality control of standard carotid endarterectomy using patch angioplasty. AB - The absence of technical defects is considered to be of great importance during carotid endarterectomy (CEA). In this context, both safe surgical technique and intraoperative quality control may be a fundamental part of the operative procedure. We have therefore undertaken a prospective study to evaluate the possible benefits of completion angiography in standard CEA using patch angioplasty. The objectives were three-fold: (1) to identify the incidence of defects requiring prompt revision; (2) to assess the perioperative stroke rate as well as the number of residual stenosis after 6 weeks in angiographically controlled patients and (3) to compare these results with a control group. From 1 January to 30 September 1999 111 patients with 115 consecutive CEAs which had completion angiography (Group A) were prospectively entered into this study. The results in group A were compared with a series of again 111 patients (Group B) which had 116 CEAs without intraoperative quality control between January and September in the year before. Surgical technique was identical in both groups. In general, risk factors were distributed evenly among both group with the exception that in group A were significantly more high-grade ipsilateral ICA stenoses while group B had more patients with diabetes and ipsilateral CT-defects. In group A, angiographic irregularities prompted us to immediate re-exploration in five patients (dilatation of severe ICA spasm 1; re-exploration of distal ICA occlusion 1; reopening of occluded ECA 3). With a 30 day mortality of 0% each perioperative stroke rate was comparable with 3/115 in group A and 3/116 in group B (P=1.0). 2/3 patients with neurological deficits in group A had early postoperative carotid thrombosis--in spite of a normal completion study. Duplex examination after 6 weeks revealed one asymptomatic ICA occlusion in each group. The incidence of residual stenosis (> or =50%) was not significantly different being 3.7% in group A and 3.2% in group B (P=0.85). When applying a safe and simple operative technique for CEA, the incidence of abnormalities warranting immediate correction appears to be a rare event and, therefore, the necessity for obligatory quality control may be questionable. On the other hand, completion DSA allows a simple documentation of the adequacy of the surgical procedure. PMID- 11888740 TI - Can cerebral vasoreactivity predict cerebral tolerance to carotid clamping during carotid endarterectomy? AB - Cerebral vasoreactivity (CVR) was evaluated as a preoperative test in predicting cerebral tolerance to carotid clamping.A consecutive series of 115 carotid endarterectomy (CEA) cases were studied. Before surgical operation CVR was evaluated, by measuring the mean velocity of the middle cerebral artery (mv-MCA) using transcranial Doppler (TCD) at the basal condition and 20 min after intravenous administration of acetazolamide (1 g). CEA was performed under general anesthesia. TCD was used during surgery to evaluate mv-MCA and to calculate mv-MCA clamping/mv-MCA pre-clamping x100 ratio (mv-MCA%), which was used as the parameter to validate CVR.CVR did not correlate with mv-MCA% (r=0.22). There was no significant difference (P=0.09) between mean values of the non shunted subgroup and the shunted one.CVR does not seem to be suitable for evaluating cerebral tolerance to carotid clamping. PMID- 11888741 TI - Splanchnic oxygenation in patients undergoing abdominal aortic aneurysm repair and volume expansion with eloHAES. AB - BACKGROUND: Gastric intramucosal pH (pHi), a surrogate marker of splanchnic oxygenation, falls following abdominal aortic aneurysm surgery. AIM: To investigate the effects of volume expansion with hydroxyethyl starch (eloHAES) on splanchnic perfusion compared to another colloid such as gelofusine. PATIENTS AND METHODS: Twenty-two consecutive patients undergoing AAA repair were randomised to receive either eloHAES or gelofusine as plasma expanders. Tissue oxygenation was monitored (10 gelofusine and 12 eloHAES) indirectly by measuring pHi using a nasogastric tonometer. RESULTS: Compared to the eloHAES group, the fall in pHi was significantly greater in the gelofusine group at clamp release (7.29 vs 7.33, P=0.003) and at 4 h following clamp release (7.29 vs 7.33, P=0.03). There was a good inverse correlation between the lowest pHi and the peak serum interleukin-6 (r(s)= -0.47, P=0.03). By multivariate analysis, the only factor that influenced the pHi was the type of colloid used (F=5.54, P=0.005). The eloHAES treated patients required significantly less colloid on the first postoperative day (3175 +/- 175 vs 4065 +/- 269 ml, P=0.01). CONCLUSION: In patients undergoing abdominal aortic aneurysm repair, plasma expansion with eloHAES improves microvascular perfusion and splanchnic oxygenation. PMID- 11888742 TI - A rabbit model of highkinetic one-sided pulmonary hypertension induced by Pott's operation. AB - This article presents a new rabbit model of pulmonary hypertension using rabbits. The left pulmonary artery of the rabbits was anastomosed on the descending aorta side-to-side and the proximal left pulmonary was banded. A portion of the arteries in the left upper lobe was ligated, and the majority of shunted blood was driven to the left middle and lower lobe. The left pulmonary hypertension was established in 86.7% of the rabbits within 1-3 months post operation and the left pulmonary hypertension was formed in all rabbits after 3 months post operation with pathological changes of pulmonary arteriole. The left pulmonary arterial pressure was higher than right pulmonary arterial pressure and the pathological changes were less evident in the right pulmonary arteriole. PMID- 11888744 TI - Hemorrhage related reexploration following open heart surgery: the impact of pre operative and post-operative coagulation testing. AB - PURPOSE: Our objective was to determine the association of pre-operative and post operative coagulation testing abnormalities with the cause of post-operative bleeding requiring re-exploration following cardiac surgery. METHODS: Retrospective chart review of post-operative bleeding and the incidence of re exploration for hemorrhage in 2263 adult patients undergoing elective and emergency open heart surgery which included coronary artery bypass, valvular, and combined valve coronary procedures. RESULTS: Eighty-two patients (3.6%) required re-exploration. Sixty-six percent had surgical bleeding; the remaining 34% were coagulopathic (no surgical site found). The pre-operative PT and ACT were significantly elevated in coagulopathic patients (P<0.005). Post-operative ACT, PT, and APTT were increased and fibrinogen levels were decreased in coagulopathic patients (P<0.05). CONCLUSIONS: Pre-operative testing (ACT, PT) weakly correlated with post-operative coagulopathy. Post-operative coagulation abnormalities were identified with high risk ratios and good diagnostic accuracy when using testing cut-off values to assist in surgical decision making. PMID- 11888743 TI - The effects of vasoactive intestinal peptide on monocrotaline induced pulmonary hypertensive rabbits following cardiopulmonary bypass: a comparative study with isoproteronol and nitroglycerine. AB - BACKGROUND: Vasoactive intestinal peptide (VIP) has regulatory effects on myocardial and vasomotor functions usually demonstrated by in vitro or isolated heart studies. We studied in vivo effects in monocrotaline induced pulmonary hypertensive rabbits immediately after cardiopulmonary bypass (CPB) and tested them versus calcium channel and beta-blockers. METHODS: The study consisted of six groups (N=30; five rabbits/group): (1) Control with no pretreatment, monocrotaline injected groups: (solutions were perfused following termination of CPB for 60 min); (2) Control for pulmonary hypertension (PHT); (3) isoproteronol; (4) VIP 10(-6) M; (5) VIP 10(-5) M; (6) nitroglycerine. Normothermic CPB was instituted in thirty rabbits at a flowrate of 100 ml/kg/min for 120 min. Heart rate, mean arterial pressure (MAP), central venous, left atrium (LAP), pulmonary artery (PAP) pressures, pulmonary resistance (Rp), blood gases and ions were measured before and 15, 30, 45 and 60 min after CPB. The VIP 10(-5) M group was subjected to an additional 1.7 x 10(-6) M propranol and 2 mM verapamil infusions for a further 15 min. RESULTS: LAP, PAP, Qp, and Rp were significantly higher in the PHT control group (P<0.001). VIP 10(-5) M increased MAP and decreased PAP significantly with respect to isoproteronol and VIP 10(-6) M (P<0.05). VIP 10(-5) M also decreased Rp significantly in the early post CPB 15th minute (P<0.05), but did not show any superiority to other agents in the following minutes. Verapamil inhibited VIP 10(-5) M effects but propranol did not. CONCLUSION: VIP has dose responsive, positive inotropic and pulmonary vasodilatory effects in whole body CPB model acting via calcium channels. PMID- 11888745 TI - Acute abdominal aortic thrombosis following the Heimlich maneuver. AB - Complications from the Heimlich maneuver are relatively infrequent. Two fatal cases of abdominal aortic thrombosis have been reported following this technique. We report on the first patient that suffered an acute thrombosis of the abdominal aorta and survived. Prompt recognition of this complication provides the only hope of survival from this rare and catastrophic complication. PMID- 11888746 TI - Successful surgical treatment of a patient with multiple visceral artery aneurysms due to fibromuscular dysplasia. AB - Multiple visceral artery aneurysms due to fibromuscular dysplasia are rare. A 43 yr-old man with a pulsatile abdominal mass detected by ultrasonography had multiple visceral artery aneurysms diagnosed by angiography. This included a huge superior mesenteric artery aneurysm. Aneurysm resection and arterial reconstruction was performed successfully. Pathologic examination revealed fibromuscular dysplasia of the medial fibroplasia type. PMID- 11888747 TI - Management of infected femoral closure devices. AB - An increase in infectious complications has been noted with the introduction of percutaneous femoral artery closure devices. We report five cases of infected groins and/or femoral arteries following angiographic procedures that were completed using the Perclose Suture Mediated Closure Device (Perclose). Each patient required drainage of the abscess and removal of the Perclose suture. Most patients required more extensive vascular reconstructive procedures. When these complications arise, we recommend expeditious drainage of the abscess, removal of the suture, and adequate exposure of the femoral artery to facilitate repair of the vessel. PMID- 11888748 TI - Abstracts of the Society for Vascular Surgery 55th Annual Meeting. PMID- 11888749 TI - Retroperitoneal approach for aortic surgery: is it worth it? PMID- 11888750 TI - Day 0 intensive care unit discharge - risk or benefit for the patient who undergoes myocardial revascularization? AB - OBJECTIVE: Day 0 intensive care unit (ICU) discharge allows to use one ICU bed for two patients. Results of this policy were analysed. METHODS: From January 1998 to June 2001, 1194 patients who had myocardial revascularization in the morning were discharged on the same day (Group 0, n=647), or one (Group 1, n=521) or many days (Group 2, n=26) after surgery. Criteria for day 0 discharge were: early extubation with at least 2h of observation, stable hemodynamic status, no significant bleeding, no arrhythmias, normal EKG and normal neurological evolution. RESULTS: Mean ICU stay was 4.0+/-1.2h in Group 0, 17.5+/-4.0 h in Group 1 and 65.8+/-46.6h in Group 2 (P(1), among Groups, <0.001; P(2), between Groups 0 and 1, <0.001). In 613 cases (94.7% of patients in Group 0) the same ICU bed was used for another patient. Postoperative in-hospital stay was 4.1+/-2.3 d in Group 0, 4.9+/-3.1 d in Group 1 and 7.4+/-6.8 in Group 2 (P(1)<0.001; P(2)<0.001). Fifteen patients (1.2%) were readmitted to the ICU, seven in Group 0 (1.1%), five in Group 1 (1.0%) and three (11.5%) in Group 2 (P(1)<0.001, P(2) n.s.), because of bleeding (five cases in Group 0, two in Group 1, none in Group 2; P(1)<0.001, P(2)), cerebrovascular accident (two cases in Group 0, none in Group 1, three in Group 2; P(1)<0.001, P(2) n.s.), acute myocardial infarction (no case in Groups 0 and 2, two in Group 1; P(1) n.s., P(2) n.s.) and acute renal failure (no case in Group 0 and 2, one case in Group 1; P(1) n.s., P(2) n.s.). Nine patients (0.8%) died (three, 0.5%, in Group 0, three, 0.6%, in Group 1 and three, 11.5%, in Group 2; P(1)<0.001, P(2) n.s.), four (one in Group 0, two in Group 1 and one in Group 2, P(1) 0.006, P(2) n.s.) in the hospital (two from cardiac and two from non-cardiac causes) and five (two in Group 0, one in Group 1 and two in Group 2, P(1)<0.001, P(2) n.s.) outside the hospital within the 30th day of surgery (one from cardiac and four from non-cardiac causes). No patient in Group 0 died from cardiac causes. CONCLUSIONS: Day 0 ICU discharge can be obtained in selected patients without an increased risk of death or of ICU readmission. The impact in terms of resource saving is striking. PMID- 11888751 TI - Hemodynamic changes during off-pump CABG surgery. AB - OBJECTIVES: The purpose of this study was to assess the patients' hemodynamics during off-pump coronary artery bypass graft (OPCABG) surgery. METHODS: Continuous monitoring of the mean systemic arterial pressure (SAP), mean pulmonary arterial pressure (PAP), mixed venous oxygen saturation (SvO(2)) and cardiac output index (COI) was done on 55 patients undergoing complete OPCAB revascularization. Hemodynamic changes were recorded at the completion of the anastomosis before releasing coronary snaring and stabilization and compared to baseline. RESULTS: The mean age of the patients was 66.4+/-9.2 years, and on average 3.3+/-0.8 grafts per patient were performed. The average SAP drop after manipulations was -8.3+/-16.9 mmHg for the left anterior descending artery (LAD), -13.5+/-19.6 mmHg for the diagonal artery (DG), -14.6+/-13 mmHg for the optuse marginal artery (OM), and -14.2+/-13.5 mmHg for the right coronary artery. This was significant for all territories (P<0.01). The PAP significantly increased in all territories except OM (LAD: 3.7+/-6.3 mmHg, DG: 4.3+/-8.6 mmHg, OM: 1.1+/-7.2 mmHg, posterior descending artery: 2.7+/-5.6 mmHg; P<0.05). Variations in COI were significant in all territories (P<0.01) but more significantly in LAD and DG territories (-15+/-3% and -13+/-9%, respectively). The SvO(2) variations were <10% for all territories and reached only borderline significance (P=0.05) in all territories except OM. All these hemodynamic changes were well tolerated by all patients. CONCLUSIONS: Manipulation of the beating heart during OPCABG surgery brings significant fluctuations in the patients' hemodynamics. Mean PAP increase and COI drop were more significant during manipulation of the anterior territories suggesting a more severe diastolic restrictive disease during anterior wall manipulation. PMID- 11888752 TI - Complete arterial coronary revascularisation using radial artery conduit for double thoracic artery inlet flow: arterial sling operation. AB - BACKGROUND: Coronary artery bypass graft surgery with arterial revascularisation of all diseased coronary vessels is considered highly efficient because arterial grafts have an excellent long-term patency compared with venous grafts. However, problems to reach the infero-lateral wall with the in situ internal thoracic arteries usually require alternative techniques. We present the first results of a new surgical principle using a free radial artery segment to complete the arterial coronary revascularisation and concomitantly connect the internal thoracic arteries. METHODS: In patients referred for coronary bypass surgery and three-vessel disease an end-to-end anastomosis of the right internal thoracic artery and the radial artery segment preceded cardiopulmonary bypass, during which side-to-side anastomoses of the radial artery segment were used to revascularise stenotic branches of the right coronary and circumflex arteries. The left internal thoracic artery was used for revascularisation of stenotic branches of the left anterior descending artery, and finally an end-to-side anastomosis of the radial artery segment to the left internal thoracic artery was performed. Coronary artery blood flow was measured in 41 patients with Doppler flow probe. RESULTS: One hundred and ninety-two coronary anastomoses (an average of 4.2 per patient) were performed in 46 patients. We measured a mean total blood flow in the arterial sling graft of 104ml/min (range 35-221ml/min), compared with 69 and 68ml/min of the single inlet right and left internal thoracic arteries, respectively (P<0.01). Flow capacities of 104 and 120ml/min of the right and left internal thoracic arteries were measured during clamp of both the aorta and the contralateral internal thoracic artery. The mean crossclamp duration was 77min (range 51-113min). Postoperative angiography demonstrated patent graft anastomoses to all coronary arteries. There were no perioperative deaths or myocardial infarctions. One patient had a minor postoperative stroke. DISCUSSION: Complete arterial revascularisation can be achieved by the arterial sling operation with an acceptable crossclamp time and a high early rate of graft patency. The double arterial inlet provides a 50% higher blood flow to the beating heart and two-fold increase in the flow reserve compared with a single inlet. Although further research including long-term follow-up of this new principle is required, the present findings seem promising and suggest that the arterial sling operation has a potential role for complete arterial coronary revascularisation. PMID- 11888753 TI - Does microalbuminuria in diabetic patients affect the postoperative course after coronary artery bypass surgery? AB - OBJECTIVES: Microalbuminuria is a predictor of microvascular disease and a marker for multiorgan damage in diabetic patients. It has been proposed that in diabetic patients who would undergo coronary artery bypass surgery (CABG), microalbuminuria is associated with poor postoperative outcome, higher incidence of early and late morbidity and mortality. METHODS: Microalbuminuria was prospectively studied preoperatively in 24-h urinary collections for 257 consecutive diabetic patients in a 2-year period. One hundred and sixty-eight patients (65.4%) were defined as microalbuminuria negative (Group A), and 89 (34.6%) were microalbuminuria positive (Group B) with respect to the cut-off point 30 mg/24 h. RESULTS: The two groups did not differ with respect to preoperative and operative data, except that preoperative blood glucose levels (P=0.046), blood urea nitrogen (P=0.001), and creatinine (P=0.001) were higher and creatinine clearance was lower (P=0.025) in Group B. Postoperative serum creatinine levels on different days were higher in microalbuminuria positive patients (P=0.04). Also, positive inotropic agent usages at the time of leaving the operating room (21.3 vs. 10.1%; P=0.013) and on the 1st day in the intensive care unit (ICU; 29.2 vs. 14.9%; P=0.014), ICU stay day (2.3+/-2 vs. 2.4+/-1.6; P=0.02) and also atrial fibrillation rate (30.3 vs. 17.9%) were higher in Group B (P=0.019). Total hospital stay (7.5+/-2.9 vs. 7.2+/-1.3) was similar. The 30-day mortality was 5.6 times higher (3.4 vs. 0.6%) but statistically not significant (P=0.088) in Group B. The mean follow-up was 30.6+/-16. 2 months in total (30.9+/ 16.2 in Group A and 30.1+/-16.5 in Group B). There were 12 late deaths, nine were cardiac, and no differences were detected between groups. CONCLUSIONS: Our findings suggest that postoperative period may be more problematic in diabetic patients with microalbuminuria, but microalbuminuria does not seem to have a major effect on the postoperative course in patients undergoing CABG. PMID- 11888754 TI - Amiodarone versus digoxin and metoprolol combination for the prevention of postcoronary bypass atrial fibrillation. AB - OBJECTIVE: This prospective randomized study aims at evaluation and comparison of the prophylactic effects of amiodarone versus digoxin and metoprolol combination in postcoronary bypass atrial fibrillation. METHODS: A total of 241 consecutive patients undergoing elective coronary artery bypass grafting were randomly allocated into three groups. Patients in Group1 (n=77) received metoprolol 100 mg/24 h per oral (P.O.), preoperatively, 2x0.5 mg digoxin intravenously on the operating day and digoxin 0.25 mg P.O.+metoprolol 100 mg P.O. on the first postoperative day until discharge. Patients in Group 2 (n=72) received totally 1200 mg intravenous/24 h amiodarone which the 300 mg - bolus dose/1 h was given as soon as the operation had been finished. On the next day patients were administered 450 mg/24 h amiodarone i.v. and 600 mg/day in three doses P.O. were given until discharge. Group 3 (n=92) was the control group with no antiarrhythmic prophylaxis. RESULTS: Preoperative patient characteristics and operative parameters were similar in three groups. Atrial fibrillation occurred in 13 patients (16.8%) in Group 1, six patients (8.3%) in Group 2 and 31 patients (33.6%) in Group 3. CONCLUSION: Both study groups were effective in the prevention of postcoronary bypass atrial fibrillation with respect to control (P<0.01 in Group 1 and P<0.001 in Group 2). PMID- 11888755 TI - Performance of three preoperative risk indices; CABDEAL, EuroSCORE and Cleveland models in a prospective coronary bypass database. AB - OBJECTIVES: The aim of the present study was to evaluate the performance of three different preoperative risk models in the prediction of postoperative morbidity and mortality in coronary artery bypass (CAB) surgery. METHODS: Data on 1132 consecutive CAB patients were prospectively collected, including preoperative risk factors and postoperative morbidity and in-hospital mortality. The preoperative risk models CABDEAL, EuroSCORE and Cleveland model were used to predict morbidity and mortality. A C statistic (receiver operating characteristic (ROC) curve) was used to test the discrimination of these models. RESULTS: The area under the ROC curve for morbidity was 0.772 for the CABDEAL, 0.694 for the EuroSCORE and 0.686 for the Cleveland model. Major morbidity due to postoperative complications occurred in 268 patients (23.6%). The mortality rate was 3.4% (n=38 patients). The ROC curve areas for prediction of mortality were 0.711 for the CABDEAL, 0.826 for the EuroSCORE and 0.858 for the Cleveland model. CONCLUSIONS: The CABDEAL model was initially developed for the prediction of major morbidity. Thus, it is not surprising that this model evinced the highest predictive value for increased morbidity in this database. Both the Cleveland and the EuroSCORE models were better predictive of mortality. These results have implications for the selection of risk indices for different purposes. The simple additive CABDEAL model can be used as a hand-held model for preoperative estimation of patients' risk of postoperative morbidity, while the EuroSCORE and Cleveland models are to be preferred for the prediction of mortality in a large patient sample. PMID- 11888756 TI - Risk factors for intestinal ischaemia in cardiac surgical patients. AB - OBJECTIVE: Mesenteric ischaemia is an uncommon (<1%) but serious complication of cardiac surgery associated with a mortality >50%. Predictors of this complication are not well defined, and diagnosis can be difficult and prompt surgical intervention can be lifesaving. METHODS AND RESULTS: In a retrospective case-note analysis from May 1994 through to May 2000, we identified mesenteric ischaemia in 39 of 5349 consecutive patients (0.07%) undergoing cardiac surgery with cardiopulmonary bypass. By logistic multivariate analysis, we have identified six possible predictors of intestinal ischaemia: duration of cross-clamp, use of significant inotropic support, intra-aortic balloon counterpulsation for low cardiac output, need for blood transfusions, triple vessel disease and peripheral vascular disease. In all patients a combination of four predictors were present. Patients who survived this complication had surgical intervention earlier (6.4+/ 3.8 h) than those who did not (16.9+/-10 h). CONCLUSIONS: The diagnosis and prompt treatment of mesenteric ischaemia post cardiac surgery requires a high degree of awareness. These predictors may be useful in alerting medical staff to the possibility of gastro-intestinal ischaemic complications after cardiac surgery particularly that early surgical intervention reduces mortality. PMID- 11888757 TI - Prolonged ischemic heart disease and coronary artery bypass - relation to contractile reserve. AB - OBJECTIVE: A major effect of coronary artery bypass grafting (CABG) in patients with ischemic heart disease and impaired left ventricular (LV) contractile function is believed to be an improvement in LV function due to recovery of dysfunctional, but viable myocardium. However, recent studies have indicated a time limit for such a recovery. We therefore investigated the extent of viable myocardium in patients with impaired LV function due to ischemic heart disease after a prolonged strategy of medical treatment and its relation to changes in clinical variables after CABG. METHODS: Forty-five consecutive patients with a mean duration of ischemic heart symptoms of 9 years and LV ejection fraction (EF) <45% referred for CABG were included and LV extent of viable myocardium was measured preoperatively by glucose metabolism--blood flow positron emission tomography imaging and dobutamine stress echocardiography. Symptoms, exercise capacity and LV function were evaluated before and 7 months after surgery in event-free survivors. RESULTS: LV extent of myocardial viability was <30% in most patients. In event-free survivors, LVEF decreased from 31+/-7 to 26+/-8% 7 months after CABG. The decrease in LVEF was correlated to the LV extent of myocardial metabolism--blood flow reverse mismatch. Most of the patients experienced an improvement in their angina pectoris, heart failure symptoms and exercise capacity after CABG; the overall 3-year survival was 77%. CONCLUSIONS: Patients with chronic ischemic heart disease and impairment of LV function, in whom an initial long-standing conservative treatment has been practiced, benefit from CABG, despite a lack of LV functional reserve. PMID- 11888759 TI - Redox behavior of cytochrome oxidase and neurological prognosis in 66 patients who underwent thoracic aortic surgery. AB - OBJECTIVE: Using near-infrared spectroscopy (NIRS), we have developed a new approach to the measurement of the redox state of cytochrome oxidase (cyt. ox.) in the brain. Our previous animal study showed that oxygen-dependent redox changes in cyt. ox. occur only when oxygen delivery is badly impaired. Therefore, in this study, we retrospectively examined the relationship between the redox behavior of cyt. ox. (measured by NIRS) during an operation and the neurological outcome in patients. METHODS: We studied 66 patients undergoing thoracic aortic surgery with cardiopulmonary bypass. Cerebral oxygenation was monitored by NIRS, and relative values for the concentrations of oxy-Hb, deoxy-Hb, and the redox state of cyt. ox. in the brain were calculated using our developed algorithm. RESULTS: Retrospective assessment revealed three different types of cyt. ox. behavior: (1) no change (type-A) in 34 cases (51.5%), (2) a temporary reduction, with a subsequent return to the pre-surgery baseline level (type-B) in 29 cases (43.9%), or (3) a marked and prolonged reduction (type-C) in only three cases (4.5%). Nine of the 66 patients (13.6%; one type-A, five type-B, and all three type-C patients) showed evidence of postoperative brain injury (in the type-A patient, the injury proved to be localized and far from the monitoring site). The relationship between the occurrence of such an injury and the type of cyt. ox. behavior seen during the operation was highly significant (P<0.0001; chi-square test for independence). CONCLUSIONS: Our data suggest that the redox behavior of cyt. ox. during an operation is a good (though not perfect) predictor of postoperative cerebral outcome, and that overall tissue oxygen sufficiency can be confirmed by near-infrared measurement of cyt. ox. PMID- 11888758 TI - Selective opening of mitochondrial ATP-sensitive potassium channels during surgically induced myocardial ischemia decreases necrosis and apoptosis. AB - OBJECTIVE: Mitochondrial ATP-sensitive potassium channels have been proposed to be myoprotective. The relevance and specificity of this mechanism in cardiac surgery was unknown. The purpose of this study was to examine the effects of the mitochondrial potassium ATP-sensitive channel opener diazoxide on regional and global myocardial protection using a model of acute myocardial infarction. METHODS: Pigs (n=19) were placed on total cardiopulmonary bypass and then subjected to 30 min normothermic regional ischemia by snaring the left anterior descending coronary artery (LAD). The aorta was then crossclamped and cold blood Deaconess Surgical Associates cardioplegia (DSA; n=6) or DSA containing 50 microM diazoxide (DZX; n=6) was delivered via the aortic root and the hearts subjected to 30 min hypothermic global ischemia. The crossclamp and snare were removed and the hearts reperfused for 120 min. RESULTS: No significant differences in preload recruitable stroke work relationship, Tau, proximal, distal or proximal/distal coronary flow, regional or global segmental shortening, systolic bulging or post systolic shortening were observed within or between DSA and DZX hearts during reperfusion. Infarct was present only in the region of LAD occlusion in both DSA and DZX hearts. Infarct size (% of area at risk) was 33.6+/-2.9% in DSA and was 16.8+/-2.4% in DZX hearts (P<0.01 versus DSA). Apoptosis as estimated by TUNEL positive nuclei was 120.3+/-48.8 in DSA and was significantly decreased to 21.4+/ 5.3 in DZX hearts. Myocardial infarct was located centrally within the area at risk in both DSA and DZX hearts but was significantly increased at borderline zones within the area at risk in DSA hearts. CONCLUSIONS: The addition of diazoxide to cardioplegia significantly decreases regional myocardial cell necrosis and apoptosis in a model of acute myocardial infarction and represents an additional modality for achieving myocardial protection. PMID- 11888760 TI - Myocardial injury in hypertrophic hearts of patients undergoing aortic valve surgery using cold or warm blood cardioplegia. AB - OBJECTIVES: Myocardial protection techniques during cardiac surgery have been largely investigated in the clinical setting of coronary revascularisation. Few studies have been carried out on patients with left ventricular hypertrophy where the choice of delivery, and temperature of cardioplegia remain controversial. This study investigates metabolic changes and myocardial injury in hypertrophic hearts of patients undergoing aortic valve surgery using antegrade cold or warm blood cardioplegia. METHODS: Thirty-five patients were prospectively randomised to intermittent antegrade cold or warm blood cardioplegia. Left ventricular biopsies were collected at 5min following institution of cardiopulmonary bypass, 30min after cross-clamping the aorta and 20min after cross-clamp removal, and used to determine metabolic changes during surgery. Metabolites (adenine nucleotides, amino acids and lactate) were measured using high pressure liquid chromatography and enzymatic techniques. Postoperative myocardial troponin I release was used as a marker of myocardial injury. RESULTS: Ischaemic arrest was associated with significant increase in lactate and alanine/glutamate ratio only in the warm blood group. During reperfusion, alanine/glutamate ratio was higher than preischaemic levels in both groups, but the extent of the increase was considerably greater in the warm blood group. Troponin I release was markedly (P<0.05, Mean+/-SD) lower at 1, 24 and 48h postoperatively in the cold compared to the warm blood group (0.51+/-0.37, 0.37+/-0.22 and 0.27+/-0.19 vs. 0.75+/ 0.42, 0.73+/-0.51 and 0.54+/-0.38ng/ml for cold vs. warm group, respectively). CONCLUSIONS: Cold blood cardioplegia is associated with less ischaemic stress and myocardial injury compared to warm blood cardioplegia in patients with aortic stenosis undergoing valve replacement surgery. Both cardioplegic techniques, however, confer sub-optimal myocardial protection. PMID- 11888761 TI - Triple valve repair for young rheumatic patients. AB - BACKGROUND: Facing young foreign polyvalvular rheumatic patients, for which long term anticoagulation is not available, we have chosen to attempt triple valve repair procedures in order to avoid prosthetic implantation in this particular population suffering from triple valve disease. METHODS: Twenty-one young rheumatic patients (mean age:11+/-4 years) underwent triple valve repair procedures including cusp extension on the aortic valve aortic between September, 1992 and December, 2000. Valvular pathology characteristics according to Carpentier's classification included mitral insufficiency type III post+II ant (n=10), type III post (n=4), type II ant (n=2), mitral stenosis (n=5), type III aortic insufficiency (n=21), type I (n=13) and type III (n=8) tricuspid insufficiency. RESULTS: Firstly, the mitral valve disease were corrected using Carpentier's techniques of repair: prosthetic ring annuloplasty (n=16), commissurotomy (n=12), chord transposition (n=11) or shortening (n=4), papillary muscle sliding plasty (n=4) and pericardial patch leaflet enlargement (n=6). Secondly, aortic lesions were corrected using glutaraldehyde stabilized autologous pericardium triple cusps extension technique (n=21). Lastly, tricuspid repairs were always performed on beating hearts using commissurotomy (n=8), prosthetic ring (n=12) or other techniques (n=9) of annuloplasty. The operative mortality was 4.7% (one patient died). Echocardiograms before discharge showed grade I mitral insufficiency in seven patients and grade I aortic insufficiency in five patients. There was no late death during a mean follow-up of 51+/-31 months. Two patients underwent valvular redo surgery because of aortic and mitral plasty deterioration due to rheumatic disease progress. After 5 years, 90% of the patients were free from redo valvular surgery. CONCLUSIONS: In rheumatic patients, autologous pericardial patch extension of the aortic valve permitted widespread use of reconstructive surgery even in patients suffering from triple valve disease. Triple valve repair, in this particular challenging setting of patients, has provided satisfactory initial and mid-term results and could be considered as an interesting palliative surgical approach. PMID- 11888762 TI - Late dissection of the ascending aorta after previous cardiac surgery: risk, presentation and outcome. AB - OBJECTIVE: Aortic dissection is a potentially life-threatening condition and may follow surgical interventions as a complication with distinct presentation and high mortality. Information on the incidence and etiology of aortic dissections following cardiac surgery is sparse and inconsistent. The true incidence of this entity may so far have been underestimated. METHODS: Data of 223 operations on the thoracic aorta performed exclusively at our institution between January 1990 and May 2001 were analysed for clinical and prognostic features. Patients with Marfan syndrome and traumatic cases were not included. Cases of type A aortic dissection following cardiac surgery were investigated further. RESULTS: Dissection of the ascending aorta occurred in 83 patients, of whom 11 (13.2%, six acute and five chronic) had undergone previous cardiac surgery (four aortic valve replacements (AVR), two double valve replacements (DVR), two AVR+coronary artery bypass grafts (CABG), three CABGs). The time interval between first operation and dissection was 0.2-17 years (median 3.3 years). Eight (72%) patients had arterial hypertension. The aortic diameter was >or=50mm in all 11 cases upon presentation. Dissections were treated with Bentall procedures (3), Cabrol procedure (1), supracoronary tube graft (6) including concomitant CABG (3) and AVR with local repair (1). Total in-hospital mortality was 54% (6/11), and 66% (4/6) in cases with acute dissection due to low cardiac output (3) and myocardial infarction (3). CONCLUSIONS: Type-A aortic dissection can follow cardiac operations at any time with no typical interval or associated histology and with high overall hospital mortality. Male patients with arterial hypertension are at increased risk. Clinical presentation may differ from primary dissection with implications for management and risk estimation. PMID- 11888763 TI - Aortic root remodeling in atheromatous aneurysms: the role of selected sinus repair. AB - OBJECTIVE: Atheromatous ascending aortic aneurysms (AAA) frequently present with aortic regurgitation (AR) from dilatation of the sino-tubular junction (STJ) and extension of the pathological process into the root. Experience suggests that root dilatation begins in the non-coronary, then right coronary sinus. Rather than employ aortic root replacement or the David procedure, we have elected to replace the ascending aorta and remodel the STJ and involved sinuses. We studied the outcome after selective sinus replacement in 29 consecutive AAA patients between 1995 and 2001. METHODS: There were nine male and 20 females. Age ranged from 47 to 79 years (mean 67.5). Seven had arch aneurysms and four coronary artery disease. Nineteen were NYHA III or IV. Grade of AR was IV in 20, III in five and II in four. The STJ was dilated >50% of annulus diameter in each case (5.3-10.0 cm, mean 6.4 cm). All valves had three cusps. All patients underwent ascending aortic replacement. Seven had arch replacement and four coronary artery bypass. Seven had replacement of both right and non-coronary sinuses with re implantation of the right coronary ostium. Twelve had replacement of the non coronary sinus alone whilst nine had right coronary sinus replacement. One with dextrocardia had left coronary sinus replacement with ostial re-implantation. The graft size was within 2 mm of annulus size except for two patients (24 mm 12, 26 mm 11, and 28 mm six). Post operative echocardiographic studies were performed. None of the patients received anticoagulation. RESULTS: There were no hospital or late deaths and no thromboembolic or infective complications. Two patients had mild to moderate aortic regurgitation. These had a size 28 graft, which in retrospect was too large. Others had no significant regurgitation. CONCLUSIONS: The native aortic valve can be preserved in the majority of patients with AAA. Remodelling of the STJ and selective sinus replacement restores valve competence. Anticoagulation and prosthesis related complications are thereby avoided. PMID- 11888764 TI - Midterm result of leaflet extension technique in aortic regurgitation. AB - OBJECTIVES: Aortic valve leaflet extension using glutaraldehyde-fixed autopericardium in aortic regurgitation (AR) is technically demanding, and it is not a popular procedure among surgeons. This study evaluates the effectiveness and clinical feasibility of the leaflet extension technique as a treatment modality for AR. MATERIALS AND METHODS: From March 1995 to March 2001, 34 consecutive patients underwent the leaflet extension technique. The mean age of the 26 men and eight women was 31.0+/-14.3 years (range 16-68). They were all diagnosed with pure aortic valve regurgitation, and nine (27.3%) had associated mitral valvular heart diseases. Accurate measurement of the leaflet free margins was performed with a pair of compasses before leaflet extension. Glutaraldehyde fixed autopericardium was designed according to the free margin lengths. Leaflet extension was performed in three aortic leaflets for 29 patients but only one leaflet was extended in the remaining four. The nine patients with associated mitral valvular heart disease also underwent mitral valvuloplasty. Mean cardiopulmonary bypass time and ischemic time in 25 isolated AR group were 128.7+/-26.5min (range 70--180) and 101.5+/-25.5min (range 41-150), respectively. RESULTS: There was no incidence of in-hospital mortality. Immediate postoperative echocardiography revealed grade II AR in one, grade I AR in ten and no AR in the remaining 23 patients. Mean follow-up duration was 49.6+/-18.8 months (range 4.1 77.1). Echocardiographic AR grades during follow-up were grade II in 13, grade I in 11 and no AR in eight. The remaining two patients underwent reoperation, one aortic valve replacement because of subacute bacterial endocarditis that occurred 7 months after leaflet extension, and the other Ross operation because of the dehiscence of the valvuloplasty suture site that developed 4 months later (AR associated with Behcet's disease). There was one case of mortality due to malignant mesothelioma 4 years after aortic valvuloplasty. The cumulative survival rate was 94.1% at 5 years. Freedom from reoperation was 93.8% at 1 year and after. CONCLUSIONS: The leaflet extension technique is an acceptable surgical treatment modality for AR and its clinical results were confirmed in this study to be very good. A careful long-term follow-up study will be necessary to evaluate the long-term durability of the glutaraldehyde-preserved autologous pericardium as a leaflet tissue. PMID- 11888765 TI - Dilatation of the autograft root after the Ross operation. AB - OBJECTIVE: Structural differences of the pulmonary root may predispose it to progressive dilatation in the systemic circulation after the Ross operation. We identified the incidence and risk factors of pulmonary autograft root dilatation. METHODS: One hundred and seven adult patients (mean age of 36+/-11 years) were followed after the Ross operation since 1991 including an echocardiogram within 3 months of surgery and yearly clinical assessment and echocardiography. The autograft was measured at the maximum diameter of the sinus (SV) and aortic insufficiency (AI) assessed. A SV of >37 mm was considered as root dilatation and the incidence over time was calculated using the Kaplan-Meier method. Clinically relevant dilatation was defined as a root diameter of >42 mm. In addition, we determined the percentage change of the sinus diameter between the early and latest echocardiogram. Furthermore we tested the influence of patient variables and risk factors on dilatation. RESULTS: By 1 year, dilatation was found in 21 patients (20%). The SV was >42 mm in eight patients (7%). By 7 years, only 45% of patients were free of dilatation. Eleven patients (10%) had a SV of >42 mm. Increase in SV was time related and linear. However, 90% of patients showed <25% dilatation during follow-up. Time from operation, early SV diameter, male gender and surgical technique were identified as significant risk factors of dilatation. However, dilatation has not lead to reoperation due to aneurysm formation or development of significant AI. CONCLUSIONS: We conclude that time dependent autograft root dilation occurs but does not cause an increase in AI and need for reoperation up to 7 years. These findings warrant the pursuit of the concept of the Ross operation in young patients who regain excellent functional status and life style without anticoagulation. PMID- 11888766 TI - Totally anomalous pulmonary venous connection directly to the superior caval vein. AB - OBJECTIVE: Totally anomalous pulmonary venous connection directly to the superior caval vein is unusual. It is frequently associated with major congenital heart defects such as the syndrome of right isomerism. While improved results have been reported recently for the isolated form, complex cases are still associated with a higher mortality. METHODS: We undertook surgical correction in nine patients with direct pulmonary venous connection to the superior caval vein investigated in our institution from 1991 to 1999. In four of these patients, the venous anomaly was an isolated finding, while five patients with isomeric right atrial appendages had associated cardiac malformations rendering them unsuitable for biventricular repair. In one patient with an isolated form, the venous drainage was obstructed. Two patients with isomerism had previously undergone construction of an aortopulmonary shunt. Other associated surgical procedures in the patients with right isomerism were establishment of bidirectional cavo-pulmonary anastomoses in four cases and banding of the pulmonary trunk in one. RESULTS: There were neither early nor late deaths. Reoperation was needed in one patient because of pulmonary venous stenosis. In the five patients with right isomerism, two later underwent successful creation of the Fontan circulation. CONCLUSION: It is unusual to find direct drainage of all the pulmonary veins to the superior caval vein. When seen, the venous pathway is only rarely obstructed. For this reason, when associated with right isomerism, an aortopulmonary shunt should be constructed as initial palliation, with later repair of the anomalous pulmonary venous drainage at the time of construction of a bidirectional cavo-pulmonary anastomosis. When using this policy, the surgical results can be as good for the complicated variant as for the isolated form. PMID- 11888767 TI - Activity-rest stimulation protocol improves cardiac assistance in dynamic cardiomyoplasty. AB - OBJECTIVE: No data have ever been published regarding cardiac assistance in demand dynamic cardiomyoplasty (DDCMP). We tested the efficacy of the Doppler flow wire in measuring beat-to-beat aortic flow velocity and evaluating cardiac assistance in demand cardiomyoplasty patients. METHODS: The technique was tested in seven patients (M/F=6/1; age=57.1+/-6.2 years; atrial fibrillation/sinus rhythm=1/6; NYHA=1.4+/-0.5). Measurements were done using a 0.018inch peripheral Doppler flow wire advanced through a 5F arterial femoral sheath. Three 1-min periods with the stimulator off and three 1-min periods with clinical stimulation were recorded. We measured peak aortic flow velocity in all beats. Latissimus dorsi (LD) mechanogram was simultaneously recorded. RESULTS: Comparison between pre-operative and follow-up data showed significantly higher values of tetanic fusion frequency (TFF) and ejection fraction at follow-up, whereas mean NYHA class was significantly lower. Statistical analysis showed an increase in aortic flow velocity not only in assisted versus rest period, but also in assisted versus unassisted beats (8.42+/-6.98% and 7.55+/-3.07%). A linear correlation was found between the increase in flow velocity and LD wrap TFF (r(2)=0.53). CONCLUSIONS: In DDCMP, systolic assistance is significant and correlated to LD speed of contraction; demand stimulation protocol maintains muscle properties and increases muscle performance. PMID- 11888768 TI - Long-term results of lung volume reduction surgery. AB - OBJECTIVE: Lung volume reduction surgery (LVRS) is effective in the short and intermediate term for the improvement of pulmonary function and subjective symptoms in selected patients with advanced emphysema. The purpose of this study was to examine the long-term functional results of LVRS and to investigate which subgroups would benefit in terms of long-term survival. METHODS: All records of the patients who underwent LVRS between 1994 and, 1998 at our hospital were reviewed. RESULTS: Eighty-eight consecutive patients underwent LVRS during the period. There were 62 men and 26 women with an average age of 56.1 years (range 34-72 years). Eleven patients with alpha1-antitrypsin deficiency were included. The perioperative mortality rate (<90 days) was 2.3% (n=2). Total lung capacity (7.5+/-0.3 l) and residual volume (4.8+/-0.3 l) at 3 years remained lower than baseline (9.2+/-0.2 l, 6.5+/-0.2 l, each) (P<0.001). The mean forced expiratory volume in 1 s (FEV(1)) at 3 years (0.86+/-0.08 l) was higher than baseline (0.78+/-0.02 l), but the difference did not reach statistical significance. The FEV(1) of the patients with alpha1-antitrypsin deficiency and of those with respiratory bronchiolitis returned to baseline at one year after LVRS and showed further deterioration. Overall survival rate at 5 years was 71.0% with the mean length of follow-up of 54.2 months. The survival difference was statistically significant between patients with preoperative FEV(1) >or=28.5% and those with FEV(1)<28.5% (P=0.0152). CONCLUSIONS: The improvement of total lung capacity and residual volume persisted long after the operation. Patients with alpha1 antitrypsin deficiency and those with bronchiolitis showed early deterioration of the lung function. Patients with higher preoperative FEV(1) had a survival benefit. The favorable long-term survival might justify LVRS for the treatment of selected patients with severe emphysema. PMID- 11888769 TI - Comparison between pulmonary and hepatopulmonary hydatidosis. AB - OBJECTIVES: To compare the clinical features and the surgical approaches between single pulmonary (SPH) and hepatopulmonary hydatidosis (HPH). METHODS: The hospital and follow-up records of 141 patients who had undergone surgery for pulmonary hydatidosis in our clinic between January 1991 and January 2001 were reviewed. Forty-nine patients (34.8%) had concomitant liver cysts in addition to the pulmonary cysts and they were regarded as HPH (Group I). The remaining 92 (65.2%) patients had SPH (Group II). Both groups were compared according to their clinical, radiological and surgical features. RESULTS: Seventeen (34.7%) male and 32 (65.3%) female patients had HPH. The mean age of the patients with HPH was significantly higher than the age of those with SPH (P<0.05) and the frequency of hepatopulmonary localization, which is contrary to single pulmonary cyst, was significantly higher in females (P<0.05). The majority (67.3%) of the cysts located in the liver were solitary. Multiple pulmonary cysts were in higher ratio in Group II, compared to Group I (45.7 vs. 22.8%) and bilateral pulmonary cyst ratio was higher in Group II, as well (26.5 vs. 13%) (P<0.05). In 14 patients (28.6%), the concomitant cysts localized in the dome of liver were extirpated via right thoracophrenotomy, and in one of them sternophrenotomy was performed. There was no statistically significant difference associated with the postoperative complications and hospital stay between groups. No recurrence and mortality were recorded in Group II. CONCLUSIONS: Multi-organ localizations (especially liver) should be examined in all patients with pulmonary hydatid cysts. HPH is more frequent in female patients over 40 years of age. The pulmonary cysts in HPH show a tendency to be bilateral and multiple. HPH should be regarded as a different entity since it can cause either economic or labour loss due to the multi operations and prolonged postoperative care. The operative strategy and approach should be different in hepatopulmonary cysts especially if they locate in the right or bilateral lung. One-session operation with the improvements of its techniques and methods should be considered in selected cases. PMID- 11888770 TI - Detection of thoracotomy-induced alterations in cell- and humoral-mediated immune response. AB - OBJECTIVE: It is well known that thoracotomy leads to several complications. In this study, effects of thoracotomy on cellular and humoral immunities have been investigated. Leukocyte counts and lymphocyte counts of 100 patients operated by thoracotomy have been determined preoperatively and on the postoperative 3rd hour, and 1st, 2nd, 3rd, 5th days. Also lymphocyte surface markers (CD3, CD4, CD8, CD4/CD8, CD19, CD16/56) and immunoglobulin levels (IgG, IgA, IgM, IgE) in 40 out of 100 patients in the preoperative period and postoperatively twice on 7th day and then in the 3rd week have been detected. MATERIALS AND METHODS: For the methodology hemocounter, flow cytometer, immunoprecipitation, and enzyme-linked immunosorbent assay were used. RESULTS: A marked increase in leukocyte count while a marked decrease in lymphocyte count has been observed after thoracotomy (P<0.001). There was not any significant alteration in levels of lymphocyte surface markers and immunoglobulins in the postoperative period (P>0.2). CONCLUSION: According to these results, leucocytosis occurred but lymphocyte count decreased in the early postoperative period. Immunoglobulin levels and subpopulation of lymphocytes were not affected from the operative stress. PMID- 11888771 TI - Decortication in chronic pleural empyema - effect on lung function. AB - OBJECTIVE: The aim of the present study is to evaluate the lung function before and after the lung decortication in patients with chronic pleural empyema (CPE). METHODS: Twenty-six patients with diagnosis of CPE were evaluated in a prospective manner by lung perfusion scintigraphy, blood gas analysis and spirometry before and 35 weeks (+/-17) after the lung decortication. RESULTS: Preoperative scintigraphy showed reduction of lung perfusion on the affected side to 24.5% (+/-12.6%) in 11 right side empyemas (predicted value 55%) and to 18% (+/-8%) in 15 left side empyemas (predicted value 45%). The postoperative measurements showed improvement in perfusion to 45.2% (+/-7.7%) in patients with right side empyema and 34.1% (+/-8.5%) with the left side affection. The preoperative vital capacity (VC) was reduced to 62.3% (+/-13.8%) of the predicted value and forced expiratory volume in 1s (FEV1) to 50% (+/-15.5%) of the predicted value. Postoperatively, slight improvement was achieved to 79.8 % (+/ 12.9%) for VC and 69.2% (+/-12.7%) for FEV1. Blood gas analysis showed decreased values in majority of the patients before operation and significant improvement in postoperative evaluation. CONCLUSIONS: Perfusion and spirometry improves significantly in patients with CPE after the lung decortication but function of the affected lung remains impaired. There was no influence of the age, gender, side of the disease, bacteriology or duration of the empyema before operation on lung function. PMID- 11888772 TI - The significance of intraoperative pleural effusion during surgery for bronchogenic carcinoma. AB - OBJECTIVES: To analyze patients submitted to thoracotomy for lung carcinoma presenting with an intraoperative pleural effusion (PE). METHODS: From 1993 to 1999, 1279 patients received thoracotomy with curative intent for primary lung carcinoma. Intraoperatively, 52 patients (4%) presented a PE >100ml which was not diagnosed preoperatively. Of these, seven patients had received preoperative transthoracic fine-needle biopsy FNB and were excluded from the analysis. In the remaining 45 patients pleural fluid cytology was undertaken. In patients with cytology-negative PE, clinico-pathologic characteristics including intratumoral vascular invasion, intratumoral perineural invasion, peritumoral lymphocytic infiltrate, visceral, parietal and mediastinal pleural involvement, pTNM and survival were analyzed and compared with our total population of lung cancer patients operated on during the same period. RESULTS: The mean amount of collected fluid was 210ml (100-450ml). Of the 45 patients with intraoperative PE, 16 (35%) received exploratory thoracotomy because of pleural carcinosis or major involvement of mediastinal structures; eight (18%) received resection of the tumor, although the cytologic examination of the pleural fluid eventually resulted positive for neoplastic cells. Median survival for the two groups was 6 and 9 months, respectively. Twenty-one patients (47%) received resection of the tumor with a cytology-negative pleural fluid. In this group, analysis of clinico pathologic characteristics revealed that squamous cell type and mediastinal pleural involvement were significantly associated with the presence of intraoperative PE (P=0.01 and P=0.05, respectively); 3- and 5-year survivals of this group were similar to those observed in our total population of resected lung cancer patients (68 and 56% vs. 54 and 42%, P=0.27). CONCLUSIONS: The presence of a PE at thoracotomy during surgery for lung carcinoma is an infrequent occurrence. In more than 50% of the cases cytology is positive and prognosis is poor. In the remaining cases, however, cytology is negative and the PE should be considered as reactive; in these patients a curative resection can be accomplished with an anticipated chance of long-term survival. PMID- 11888773 TI - Stage I non-small cell lung carcinoma: really an early stage? AB - OBJECTIVE: We review our results on surgical treatment of patients with stage I non-small cell lung carcinoma and we attempted to clarify the prognostic significance of some surgical--pathologic variables. METHODS: From 1993 to 1999, 667 patients received curative lung resection and complete hilar and mediastinal lymphadenectomy for non-small cell lung cancer. Of these, there were 436 Stage I disease (65%), of whom 144 T1N0 and 292 T2N0. No patients had pre- or postoperative radio- or chemotherapy. Prognostic significance of the following independent variables was tested using univariate (log-rank) and multivariate (Cox proportional-hazards) analysis: type of resection (sublobar vs lobectomy vs pneumonectomy), histology (squamous cell vs adenocarcinoma), tumour size (3cm), histologic vascular invasion, visceral pleura involvement, positive bronchial resection margin, general T status. RESULTS: Overall 5-year survival was 63%. In both univariate and multivariate survival analysis, significant prognostic factors were histology (adenocarcinoma 65% vs squamous cell carcinoma 51%), tumour size (3cm 46%), and the presence of negative resection margin. Five-year survival by general T status was 66% in T1N0 vs 55% in T2N0 disease (P=0.19). CONCLUSIONS: Despite advances in early diagnosis and surgical technique, 5-year survival of stage I non-small cell lung carcinoma remains low as compared to survival of other solid organ neoplasm. Tumour size 4 mm, 30.6%; erythema, 48.9%; change in vascularity, 37.4%; labial adhesions, 15.6%; posterior hymenal notch/cleft (partial), 18.3%; posterior notch/cleft (complete), 0%; posterior hymenal concavity or angularity, 29.5%. In addition, each case was assessed for the presence of a thickened (45.5%) or irregular (51.7%) and narrowed (22.4%) hymenal edge. Each case was also reviewed for exposed intravaginal anatomy (93%). CONCLUSIONS: The authors concluded that improved techniques and photo documentation have provided examiners with a better understanding of hymenal morphology and that nonspecific genital findings are commonly found in a population of girls selected for nonabuse. A thorough understanding of normal studies and a consistent application of established terminology can prevent the misinterpretation of nonspecific or congenital findings as posttraumatic changes. PMID- 11888809 TI - Low parental monitoring predicts subsequent pregnancy among African-American adolescent females. AB - STUDY OBJECTIVE: Accumulating evidence suggests that parental monitoring is associated with adolescents' sexual risk behaviors. However, evidence associating low parental monitoring with greater odds of becoming pregnant has not been reported. The objective of this study was to prospectively assess the relationship of low perceived parental monitoring with incidence of biologically confirmed pregnancy among a sample of low-income African-American adolescent females. DESIGN: A prospective study. SETTING AND PARTICIPANTS: African-American females 14-18 years of age were recruited from schools and health clinics in low income neighborhoods. Adolescents completed an in-depth survey and interview at baseline and 6 months later. The study achieved an 85.7% baseline participation rate (n = 522) and 92% (n = 482) returned at follow-up. Only adolescents who initially tested negative for pregnancy were included in the analysis (n = 410). MAIN OUTCOME MEASURES Incidence of biologically assessed pregnancy. RESULTS: In controlled analyses, among adolescents testing negative for pregnancy at baseline, those who reported less parental monitoring were 2.5 times more likely to become pregnant in the 6-month follow-up period (AOR = 2.50, 95% CI = 1.1-5.9, P <.04). CONCLUSION: Low parental monitoring was prospectively associated with incidence of biologically confirmed pregnancy among minority adolescent females. This finding adds to a growing body of empirical literature that supports the value of parental monitoring as a protective factor in adolescents' lives. Interventions designed to increase parental monitoring or adolescent females' perceptions of their parents' monitoring may be effective components of pregnancy prevention programs designed for minority youth. PMID- 11888808 TI - Use of ligase chain reaction for laboratory identification of Chlamydia trachomatis and Neisseria gonorrhoeae in adolescent women. AB - STUDY OBJECTIVE: To compare the ligase chain reaction (LCR) with culture for the detection of Neisseria gonorrhoeae (GC) and with culture and direct fluorescent antibody (DFA) for identification of Chlamydia trachomatis (CT) in cervical specimens from adolescent women. DESIGN: A prospective study of test performance. SETTING: Two urban, hospital-based adolescent clinics. PARTICIPANTS: Adolescent women aged 12-22 yr undergoing pelvic examination for routine sexually transmitted disease (STD) screening or symptoms suggestive of an STD. MAIN OUTCOME MEASURES: LCR results were considered to be true positives if confirmed by culture and/or DFA (CT only). Discrepant LCR results were confirmed by testing an alternative locus. RESULTS: With 538 subjects, LCR for CT had a sensitivity of 98.4% (61/62) and specificity of 96.4% (459/476) prior to resolution and a sensitivity of 98.6% (70/71) and specificity of 99.6% (459/461) after resolution. With 1225 subjects, LCR for GC had a sensitivity of 90.0% (54/60) and specificity of 99.4% (1158/1165) prior to resolution and a sensitivity of 90.6% (58/64) and specificity of 100% (1158/1158) after resolution. CT culture alone identified 80% of the true positives and DFA alone only identified 72%. GC culture alone identified 94% of the true positives. CONCLUSIONS: LCR is an extremely sensitive and specific rapid test, utilizing a single swab and convenient room-temperature storage and transport of specimens. LCR testing of cervical specimens for CT in adolescent women is a better test for detecting CT infection than culture or DFA. LCR testing for cervical GC infection may provide an advantage over culture in circumstances in which optimal transport conditions and viability of the organism cannot be assured. PMID- 11888810 TI - Ovarian neoplasias in children. PMID- 11888811 TI - Management quandry. Vaginal bleeding and abdominal pain. PMID- 11888812 TI - Perspectives on pediatric and adolescent gynecology from the allied health professional. PMID- 11888814 TI - Echocardiographic assessment of ventricular wall motion in patients with poor acoustic windows: the uneasy transition from art to science. PMID- 11888815 TI - Point-of-care echocardiography: asking the right questions. PMID- 11888816 TI - Diastolic filling and pressure imaging: taking advantage of the information in a colour M-mode Doppler image. AB - The ability to derive non-invasively information on left ventricular diastolic function on one hand and pressure gradients on the other hand, makes Doppler ultrasound a very attractive tool in clinical practice. However, the limitations of the standard Doppler approaches in differentiating between normal and pseudonormal filling patterns, together with the limitations of the simplified Bernoulli equation for assessing pressure gradients, are well described. In this manuscript the role of colour M-mode Doppler echocardiography as a tool that can overcome these limitations is discussed. Relevant key concepts of the haemodynamics of left ventricular filling and its relationship with colour M-mode Doppler echocardiography are introduced. PMID- 11888817 TI - Intracardiac echocardiography. AB - This article describes currently available intracardiac ultrasound (ICE) technology contrasting it with intravascular ultrasound (IVUS), highlighting their differences. General and specific clinical applications, limitations and future developments of ICE are addressed. ICE is possible because lower frequency transducers (in contrast to higher frequency IVUS devices) have been miniaturized and mounted onto catheters capable of percutaneous insertion into the heart. Since the recent availability of a steerable, 5.5--10MHz phased-array catheter with full Doppler capability, these lower frequency transducers are not only capable of enhanced penetration, permitting high-resolution two-dimensional (2D) imaging but can also provide haemodynamic data. ICE facilitates electrophysiologic procedures by guiding trans-septal catheterization, enabling endocardial anatomy visualization, ensuring ablation electrode/tissue contact and promptly diagnosing procedural complications. Promising non-electrophysiologic applications include guidance of percutaneous closure of septal defects, percutaneous mitral balloon valvuloplasty and complex cardiac biopsy. Current limitations include monoplanar imaging and narrow field of view. Expanded diagnostic techniques such as tissue Doppler, multiplane, three dimensional (3D) and multimodality imaging represent future refinements. ICE is now a clinical tool. With the introduction of the newest phased-array transducer, with full Doppler capability, ICE has the potential to play an important role in diagnostic and therapeutic interventional procedures. Further refinement and miniaturization hold the key to primary operator controlled, integrated ultrasound-guided interventional devices. PMID- 11888818 TI - Impact of harmonic imaging and transducer frequency on 'ventricular volume' measurements using real-time three-dimensional echocardiography: studies in an in vitro model. AB - AIMS: Harmonic imaging has increased the yield of quantifiable scans in two dimensional echocardiography. Although real-time three-dimensional echocardiography avoids geometric assumptions in volume analysis, accurate measurement can be limited by image quality. This study compared volumes from a balloon model mimicking the left ventricle, scanned with and without harmonic imaging, using real time three-dimensional echocardiography. METHODS: Two balloons separated by ultrasound gel were suspended in a water bath. To mimic different chamber volumes, 12 volumes of water within the inner balloon (40 180ml) were scanned using a 3.5MHz probe at fundamental frequency and using a 2.5MHz probe with and without harmonic imaging. RESULTS: Scanning at 3.5MHz, the long axis (B) scans did not significantly underestimate the balloon volume but the 'short axis' (C) scans did (mean difference from actual volumes -0.7-1.4ml, P=0.14 - 3.9 +/- 1.2 ml,P < 0.0001 for B and C scans, respectively). Scanning at 2.5MHz both B and C scans significantly underestimated even more the true volume, C scans to a greater extent (mean difference -6.9 +/- 2.4ml and -11.2 +/- 4.0ml for B and C scans respectively,P < 0.0001 in both cases). However with harmonic imaging, transmitting at 1.7MHz and receiving at 2-4MHz, there was no significant difference of either B or C scans from the reference values (mean difference of B scans -1.2 +/- 1.9ml, P=0.06 and C scans -0.6 +/- 2.2ml, P=0.4). CONCLUSION: The enhanced resolution provided by harmonic imaging improves accuracy of volume analysis by real-time three-dimensional echocardiography. PMID- 11888819 TI - Quantification of left ventricular function with contrast-enhanced harmonic colour Doppler and a semiautomated boundary detection algorithm in technically difficult patients: feasibility, accuracy, and inter-observer variability. AB - AIMS: Patients with poor quality echocardiograms impede the application of available automatic boundary detection technologies. Tissue harmonic imaging and contrast media can allow optimal differentiation of left ventricular blood pool and tissue, making it possible to utilize automatic boundary detection software for automatic non-operator-dependent computation of left ventricular volumes. We integrated contrast-enhanced harmonic colour Doppler with a semiautomated boundary detection algorithm to explore the feasibility, accuracy and inter observer variability for left ventricular volume assessment in technically difficult patients. METHODS AND RESULTS: Twenty-six patients with more than two segments not clearly visualized in tissue harmonic imaging were studied with contrast-enhanced harmonic colour Doppler using Levovist. Twenty patients (77%) achieved full left ventricular contrast filling without apparent blooming artefacts. Contrast-enhanced harmonic colour Doppler-automatic boundary detection was successfully implemented in these 20 patients, despite three (15%) in which it was not possible to acquire more than three cardiac cycles' values. Contrast enhanced harmonic colour Doppler-automatic boundary detection measurements agreed closely with the manually drawn data. Among the three independent readers in the three techniques, the best correlation, lowest SEE, smallest limits of agreement and inter-observer variability were obtained in contrast-enhanced harmonic colour Doppler-automatic boundary detection. CONCLUSION: Contrast-enhanced harmonic colour Doppler-automatic boundary detection was feasible and accurate in estimation of left ventricular volume and function in patients with poor acoustic windows. This technique significantly reduced inter-observer variability, thus improving reliability and confidence of investigators in left ventricular function assessment. Contrast-enhanced harmonic colour Doppler-automatic boundary detection may have great potential in clinical evaluation of left ventricular volume and function, especially when on-line software is available. PMID- 11888820 TI - Rapid assessment of cardiac anatomy and function with a new hand-carried ultrasound device (OptiGo): a comparison with standard echocardiography. AB - AIMS: The aim of this study was to evaluate image quality and accuracy of a new hand-carried ultrasound device, OptiGo (Agilent Technologies) when compared to standard echocardiography in the setting of a focused examination in the assessment of cardiac anatomy and function. METHODS AND RESULTS: One-hundred and twenty-one patients were prospectively enrolled. Image quality and accuracy in assessment of chamber sizes, left ventricular (LV) wall thickness and contractility, right ventricular (RV) function, mitral and aortic leaflet thickening, mitral annular calcification, pericardial effusion and valvular regurgitation were assessed. Two-dimensional (2D) findings were graded on a four point scale, except for LV function (six-point) and valvular leaflet opening (two point). Colour Doppler assessment of valvular regurgitation was graded on a seven point scale. A one-point difference was considered minor; a two or more point difference was considered major. There was no statistically significant difference in image quality between the two devices. For 2D data, the number of total (minor and major) differences between the hand-carried and standard echocardiograph examinations was significantly greater than the inter-observer variability (14.3% vs 10.7%, P< 0.05), however, major differences alone were not statistically different. For the colour Doppler assessment of regurgitation there was a significant difference between the devices for total (minor and major) differences, (40.0% vs 31.8%,P < 0.007) however, the number of major differences is explained by inter-observer variability. CONCLUSIONS: Image quality and diagnostic accuracy of the hand-carried device, OptiGo, was adequate for the purpose of performing a focused assessment of a limited number of 2D and Doppler parameters for the evaluation of cardiac anatomy and function. PMID- 11888821 TI - Location, size and morphological characteristics of left atrial thrombi as assessed by echocardiography in patients with rheumatic mitral valve disease. AB - AIMS: This study aimed to assess the use of transthoracic and transoesophageal echocardiography in diagnosing the thrombi located in the left atrium and/or left atrial appendage in patients with rheumatic mitral valve disease, and to investigate the characteristics of thrombi in comparison to intraoperative findings. METHODS AND RESULTS: The study group was comprised of 474 patients who underwent transthoracic and transoesophageal echocardiography prior to mitral valve surgery. Location, thickness and morphological characteristics of thrombi were determined by transoesophageal echocardiography. Intraoperative assessment disclosed left atrial thrombi in 105 patients. Thickness of thrombi < or = 1cm, and thrombi confined to left atrial appendage were associated with false-negative results by transthoracic echocardiography. However, diameter and morphological characteristics of thrombi, left atrial and left atrial appendage size, and the presence of the spontaneous echo contrast were not associated with the diagnosis of thrombi by transthoracic echocardiography. For overall left atrial thrombi, sensitivity and specificity of transthoracic echocardiography were 32%, and 94%, respectively. Sensitivity and specificity of transoesophageal echocardiography for thrombi in the left atrial appendage were 98%, and 98%, for thrombi in the main left atrial cavity were 81%, and 99%, and for thrombi located in both left atrium and appendage cavities were 100%, and 100%, respectively. CONCLUSION: In patients with rheumatic mitral valve disease, detection of left atrial thrombi by transthoracic echocardiography seems to be determined by thickness and location of thrombi. The multilobed structure of the left atrial appendage and artifacts over posterior wall of the left atrium may still prevent precise diagnosis even with transoesophageal echocardiography. PMID- 11888822 TI - Accurate determination of mitral regurgitation by assessing its influence on the combined diastolic mitral and pulmonary venous flow: just 'looking twice'. AB - AIMS: To assess the diagnostic accuracy of combined transmitral E wave velocity and reversed systolic pulmonary venous flow for the quantification of mitral regurgitation severity. METHODS AND RESULTS: Measuring forward and total left ventricular stroke volume, mitral regurgitation severity was assessed quantitatively by calculating the regurgitant fraction in 106 consecutive patients with pure mitral regurgitation. According to the regurgitant fraction, the optimal E wave velocity for accurate distinction of mild to moderate and more than moderate mitral regurgitation was chosen by calculating the receiver operating characteristic plot. Severe mitral regurgitation was defined by reversed systolic pulmonary venous flow. Combining transmitral E wave velocity and reversed systolic pulmonary venous flow had an overall accuracy of 78% (95% CI 70--86%) for classification of mitral regurgitation severity. E wave velocity >1.0ms(-1) predicted more than moderate mitral regurgitation with 78% sensitivity (95% CI 69-86%) and 90% specificity (95% CI 82--95%), resulting in a positive likelihood ratio of 8.1 (95% CI 5--15) and negative likelihood ratio of 0.25 (95% CI 0.18--0.35). For reversed systolic pulmonary venous flow in the presence of increased E wave velocity, the sensitivity and specificity to detect severe mitral regurgitation was 78% (95% CI 69--86%) and 97% (95% CI 92--99%) with the corresponding positive and negative likelihood ratio of 29 (95% CI 11--96) and 0.22 (95% CI 0.14--0.31). Test accuracy was independent of systolic function in a multivariate regression analysis. CONCLUSIONS: 'Looking twice', once at the transmitral E wave velocity and once at pulmonary venous flow in patients with mitral regurgitation, allows accurate determination of moderately severe and severe mitral regurgitation. PMID- 11888824 TI - Echocardiographic detection of coronary artery fistula into the pulmonary artery. AB - A rare case of left coronary artery--pulmonary artery fistula is reported. Transoesophageal echocardiography was capable of precisely demonstrating the origin, the course and the drainage site of the fistula. In contrast, transthoracic echocardiography could visualise a drainage flow in the pulmonary artery only. In conclusion, transoesophageal echocardiography may be helpful in the diagnosis or exclusion of the form of coronary artery fistula. PMID- 11888823 TI - Is contrast-related vasodilatation after intra-coronary iodixanol and iopromide in vivo endothelium-dependent? AB - AIMS: Goals of the study were the assessment of the correlation between flow dependent and contrast-related vasodilatation, comparison of iodixanol to iopromide and evaluation of the impact of plaque on vasodilatation in coronary arteries. METHODS AND RESULTS: A controlled randomized paired cross-over comparison between iodixanol (320mgI.ml(-1)) and iopromide (300mgI.ml(-1)) was performed in 10 consecutive patients. Vessel area (Visions Five-64 F/X intra vascular ultrasound-catheter, Endosonics and blood flow velocity measurements (0.014inches Doppler guide wire, Cardiometrics were recorded simultaneously at an identical position, at baseline, after i.c. bolus injection of 10ml physiologic saline (flow-dependent vasodilatation), and after application of contrast agent 1 and contrast agent 2 as randomized. The action of iodixanol and iopromide on the vascular wall did not differ and was equal to local flow-dependent vasodilatation induced by a saline bolus (correlation 0.95-- 0.98). The increase in local luminal area after injection of saline, iodixanol and iopromide in morphologically normal vessels (approximately 2.5mm(2)) was absent in atherosclerotic segments. Both contrast agents and saline demonstrated a nearly identical flow increase. CONCLUSION: If iodixanol or iopromide are used as contrast agents, contrast-related vessel area increase in vivo seems to be endothelium-dependent. PMID- 11888825 TI - Transoesophageal echocardiographic visualization of renal cyst mimicking aortic aneurysm. AB - We report a tricky case of a 63-year-old man with previous coronary artery bypass graft surgery in whom transoesophageal echocardiography revealed a voluminous echolucent cavity simulating aortic ectasia but that proved to be of nephrogenic origin. PMID- 11888826 TI - Abstracts from the workshops: Frontiers of Skeletal Biology, 9th Workshop on Cell Biology of Bone and Cartilage in Health and Disease, Davos, Switzerland, March 16 19, 2002 and What is New in Bisphosphonates? 6th Workshop on Bisphosphonates from the Laboratory to the Patient, Davos, Switzerland, March 20-22, 2002. PMID- 11888831 TI - Post-partum thyroiditis--a clinical update. AB - Since the late 1960s, many studies have focused on post-partum thyroiditis (PPT) and 295 papers whose titles mention PPT were recorded on MEDLINE as of August 2001. We refer briefly to some excellent reviews and some original articles in order to update our knowledge on PPT. PMID- 11888832 TI - A molecular switch for parathyroid cell differentiation. PMID- 11888833 TI - Managing toxic thyroid adenoma: a cost-effectiveness analysis. AB - OBJECTIVE: To examine the cost-effectiveness of therapeutic strategies for patients with toxic thyroid adenoma. DESIGN: A decision analytic model was used to examine strategies, including thyroid lobectomy after a 3-month course of antithyroid drugs (ATDs), radioactive iodine (RAI), and lifelong ATDs followed by either RAI (ATD-RAI) or surgery (ATD-surgery) in patients suffering severe drug reactions. METHODS: Outcomes were measured in quality-adjusted life years. Data on the prevalence of co-incident thyroid cancer, complications and treatment efficacies were derived from a systematic review of the literature (1966-2000). Costs were examined from the health care system perspective. Costs and effectiveness were examined at their present values. Discounting (3% per year), variations of major cost components, and every variable for which disagreements exist among studies or expert opinion were examined by sensitivity analyses. RESULTS: For a 40-year-old woman, surgery was both the most effective and the least costly strategy (Euro 1391),while ATD-RAI cost the most (Euro 5760). RAI was more effective than surgery if surgical mortality exceeded 0.6% (base-case 0.001%). RAI become less costly for women of more than 72 years (more than 66 in discounted analyses). For women of 85, ATD-RAI may be more effective than RAI and have an inexpensive marginal cost-effectiveness ratio (Euro 4975) if lifelong follow-up results in no decrement in quality of life. CONCLUSIONS: Age, surgical mortality, therapeutic costs and patient preference must all be considered in choosing an appropriate therapy. PMID- 11888834 TI - Partial tachyphylaxis to somatostatin (SST) analogues in a patient with acromegaly: the role of SST receptor desensitisation and circulating antibodies to SST analogues. AB - OBJECTIVE: Somatostatin (SST) analogues are a key option in the management of a variety of conditions, including acromegaly. Tachyphylaxis to SST analogues is not documented in acromegaly. We describe such a phenomenon. DESIGN AND METHODS: A 74-year-old female with acromegaly previously treated with (90)Y implant, external radiotherapy and thrice daily s.c. octreotide had stable GH levels of 19 mU/l. GH progressively rose following switches to lanreotide and depot octreotide as Sandostatin LAR: from 29 to 126 mU/l. Magnetic resonance imaging and (111)In pentetreotide scanning revealed no tumour growth or alteration in SST receptor (SSTR) status. Tachyphylaxis to SST analogues was considered. Therapy was discontinued and re-introduced in daily 200 microg/24 h increments by continuous s.c. infusion, to a maximum of 1000 microg/24 h, and maintained over 3 weeks with daily, followed by weekly, GH profiles. Competitive (125)I-octreotide radioligand binding assays measured in vitro bio-activity of anti-SST analogue antibodies. In vitro SSTR binding studies utilised SSTR-expressing rat cortex membrane. RESULTS: Median GH fell by 93% from 504 to 39.5 mU/l and rose reproducibly on continued infusion to 120 mU/l. Octreotide withdrawal for 16 h produced a 64% increase in sensitivity. High-affinity IgG anti-lanreotide (IC(50)=187 pmol/l) and anti octreotide (IC(50)=82 nmol/l) antibody, with no crossreactivity with natural SST, was demonstrated. In vitro inhibition of (125)I-octreotide SSTR binding by anti SST analogue crossreacting antibody was observed at 1:1 serum dilution. CONCLUSIONS: This is the first report of tachyphylaxis to SST analogues in acromegaly. We believe that the short time course of resensitisation following acute octreotide withdrawal is suggestive of an effect(s) on receptor function or on the receptor signal transduction cascade at sites further downstream, rather than an immune-mediated phenomenon. PMID- 11888835 TI - Effect of a six-month treatment with lanreotide on cardiovascular risk factors and arterial intima-media thickness in patients with acromegaly. AB - OBJECTIVE: To evaluate the effect of a 6-month treatment with slow-release lanreotide (LAN) on cardiovascular risk and atherosclerosis in 24 normotensive patients with active acromegaly (GH=67.4 +/- 12.6 mU/l, IGF--I=866.0 +/- 55.8 microg/l) and 24 healthy subjects sex-, age- and body mass index-matched with the patients (as controls). DESIGN: Open, prospective, multicenter. METHODS: The following were measured before and after 6 months of LAN treatment (dose 60-90 mg/month): fasting GH, IGF-I, LDL, HDL and total cholesterol, triglyceride, glucose, glycosylated hemoglobin, insulin and fibrinogen levels, intima-media thickness (IMT) and blood systolic and diastolic peak velocity (SPV and DPV respectively) in both common carotids. RESULTS: At study entry, insulin, total and LDL cholesterol, triglyceride and fibrinogen levels were higher while HDL cholesterol levels were lower in patients than in controls. At the right (0.88 +/ 0.04 vs 0.77 +/- 0.03 mm, P=0.05) and left (0.93 +/- 0.03 vs 0.78 +/- 0.02 mm, P=0.01) common carotid IMT was significantly higher in patients than in controls; 12 patients and two controls showed an IMT of > or = 1 mm (chi(2)=8.2, P=0.004). After 6 months of LAN treatment, disease control was achieved in 15 patients (62.5%). Insulin, triglyceride and fibrinogen levels were significantly decreased, and a trend toward a decrease of IMT in the right (from 0.90 +/- 0.05 to 0.78 +/- 0.04 mm, P=0.06) and left (from 0.95 +/- 0.04 to 0.84 +/- 0.04 mm, P=0.06) common carotid arteries was observed only in patients with disease control, while SPV and DPV did not change. CONCLUSIONS: LAN treatment for 6 months significantly lowered GH, IGF-I, insulin and fibrinogen levels and reduced IMT of both common carotid arteries in normotensive patients with acromegaly. PMID- 11888836 TI - Impact of two or three daily subcutaneous injections of hexarelin, a synthetic growth hormone (GH) secretagogue, on 24-h GH, prolactin, adrenocorticotropin and cortisol secretion in humans. AB - OBJECTIVE: To extend the insights on the action of GH secretagogues (GHS) on pituitary function, we studied the impact of intermittent daily s.c. administration of a peptidyl GHS, hexarelin (HEX), on 24-h GH, PRL, ACTH and cortisol release in healthy volunteers. DESIGN: We investigated the impact of two or three times daily s.c. administration of a short-acting peptidyl GHS, the hexapeptide HEX (1.5 microg/kg) on 24-h GH, PRL, ACTH and cortisol secretion (sampling every 20 min) in six normal young men. To monitor possible down regulation, the effect of 1 microg/kg i.v. HEX at the end of each 24-h sampling period was studied. METHODS: Multi-parameter deconvolution analysis was used to quantitate pulsatile GH, PRL, ACTH and cortisol secretion and estimate the corresponding hormone half-lives. Complementary to deconvolution analysis, approximate entropy was used as a scale- and model-independent statistic to quantify the serial orderliness or pattern regularity of hormone measurements. RESULTS: Mean and integrated (24-h) serum GH concentrations were increased from baseline values to the same extent by two and three HEX injections. Both HEX schedules equally increased GH secretory burst mass (but not burst frequency), mean daily GH production rate, GH half-life and irregularity of GH release patterns. No change occurred in the secretion of IGF-I, PRL, ACTH and cortisol. Intravenous HEX at the end of each spontaneous 24-h profile induced a significant rise in GH, PRL, ACTH and cortisol. Prior HEX administration blunted the GH response, abolished that of ACTH and cortisol and did not modify the PRL increase. CONCLUSIONS: The study showed that two or three daily s.c. injections of HEX augmented 24-h GH secretion equally, amplifying selectively GH secretory pulse mass without altering lactotroph and corticotroph secretion. IGF-I levels were not modified by these 1-day HEX treatment schedules. PMID- 11888837 TI - Can exaggerated response to a GH provocative test identify patients with partial GH insensitivity syndrome? AB - OBJECTIVES: In the majority of children with short stature, the etiology is unknown. Mutations of the GH receptor (GHR) have been reported in a few children with apparent idiopathic short stature (ISS). These patients had low IGF-I, IGF binding protein-3 (IGFBP-3) and GH-binding protein (GHBP), but a normal or exaggerated GH response to provocative stimuli, suggestive of partial GH insensitivity (GHI). We attempted to identify children with partial GHI syndrome, based on their response to GH provocative stimuli and other parameters of the GH IGF-I axis. SUBJECTS AND METHODS: One hundred and sixty-four pre-pubertal children (97 boys, 67 girls) aged 7.2 (0.5-16.75) years were studied. All had short stature with height <3rd centile. The weight, bone age (BA) and body mass index (BMI) of the subjects, as well as the parents' heights and mid parental height (MPH) were assessed. Basal blood samples were taken for IGF-I, IGFBP-3 and GHBP. All subjects underwent a GH provocative test with either clonidine, arginine or insulin. The subjects were divided into three groups: (A) patients with peak GH concentration <18 mIU/l in two different provocative tests (GH deficiency - GHD, n=33); (B) patients with peak GH between 18.2 and 39.8 mIU/l (normal response, n=78); (C) patients with peak GH >40 mIU/l (exaggerated GH response, n=53). RESULTS: No significant differences were found in age, height (standard deviation score (SDS)), parental height (SDS) and the difference between chronological age and bone age (DeltaBA) between the groups. Patients with GHD were heavier (P=0.039) and had significantly higher BMI (SDS) (P=0.001) than the other groups. MPH (SDS) was lower in the group of exaggerated responders (P=0.04) compared with the other groups. No significant differences were found between the groups for the biochemical parameters when expressed nominally or in SDS, except for IGFBP-3 (SDS), which was lower in the GHD group (P=0.005). The GHBP levels were not lower in the group of exaggerated GH response to provocative stimuli. Height (SDS) correlated negatively with basal GH values in pooled data of all the subjects (r=-0.358, P<0.0001), in normal responders (r=-0.45, P<0.0001) and in the exaggerated responders (r=-0.341, P<0.0001), but not in the GHD group. CONCLUSION: Exaggerated GH response to provocative tests alone does not appear to be useful in identifying children with GHI. PMID- 11888838 TI - Plasma soluble tumor necrosis factor alpha receptors and leptin levels in normal weight and obese women: effect of adiposity and diabetes. AB - OBJECTIVE: To explore the determinants of the tumor necrosis factor alpha (TNFalpha) system and their relationship with plasma leptin levels. METHODS: We studied a cohort of 157 diabetic and non-diabetic females with a wide range of adiposity distributed into five groups: control--body mass index (BMI) between 19 and 27 kg/m(2) (n=24); obese--BMI between 27 and 40 kg/m(2) (n=63); obese type 2 diabetes mellitus--BMI between 27 and 40 kg/m(2) with diabetes mellitus (n=19); morbid obese--BMI >40 kg/m(2) (n=29); and morbid obese type 2 diabetes mellitus- BMI >40 kg/m(2) with diabetes (n=22). Fasting glucose levels, plasma total triglycerides and cholesterol, high-, low- and very low-density lipoprotein cholesterol were assayed by enzymatic and colorimetric methods. Plasma TNFalpha levels were measured by ELISA assay and insulin and leptin levels by radioimmunoenzymatic assays. Both soluble TNFalpha (sTNFalpha) receptors were measured by immunoenzymometric assays. RESULTS: All groups of patients showed significant increases in both sTNFalpha receptors relative to control. sTNFalpha receptor 1 (sTNFR1) was higher in morbid obese diabetic individuals compared with their non-diabetic counterparts (P=0.003), while sTNFR2 was significantly different between obese and morbid obese subjects (P=0.036). Bivariate correlation analysis showed a significant relationship between both plasma sTNFalpha receptors and BMI, percentage of body fat, fasting glucose, insulin and leptin. In multivariate analysis, both sTNF receptor plasma levels were predicted by percentage of body fat and the presence of diabetes (R(2)=0.20 for sTNFR1 and sTNFR2). When plasma leptin levels were added into the model, this protein and the presence of diabetes explained 27% of the variance of the plasma sTNFR1 levels. CONCLUSION: The presence of diabetes, adiposity or leptin levels are independent determinants of both sTNFalpha receptors. The independent association between plasma TNFalpha receptors and leptin levels in obese patients is consistent with the hypothesis that these proteins could be involved in the same pathway that regulates body adiposity. PMID- 11888839 TI - Self-adjusted postmenopausal hormone replacement therapy: effects on the biological and immunological profile of FSH and correlation to climacteric symptoms. AB - OBJECTIVE: The purpose of the present study was to evaluate the hormonal profile of patients of postmenopausal age during estrogen replacement therapy (ERT) with special reference to the serum levels of biologically active FSH (B-FSH) in a self-adjusted ERT model. DESIGN: The hormonal values found have been correlated to climateric symptoms reported by the patients (scored by the Kupperman menopausal index (KI)). METHODS: B-FSH was measured using an assay based on a cell system transfected permanently with FSH receptor cDNA. All women (n=32) applied estradiol percutaneously using 1 mg estradiol-17beta (E(2)) as an initial dose and were encouraged to increase the daily dose until they felt comfortable according to a specific scheme. Twelve of the 32 women were hysterectomized and treated, accordingly, with ERT only; 20 women received megestrol acetate monthly for 10 days. RESULTS: The initial average KI was 30 (range 10-54). A high degree of correlation (r=0.83; P<0.001) was observed between B-FSH and immunologically active FSH (I-FSH). Serum I-FSH and E(2) correlated negatively (r=-0.21; P<0.001); similarly, a negative correlation (r=-0.15; P<0.01) was observed between serum B-FSH and E(2) levels. Serum I-FSH and KI showed modest but significant positive correlation (r=0.13; P<0.01); a somewhat higher degree of correlation (r=0.19; P<0.005) was observed when B-FSH and KI were compared. E(2) showed positive correlation to serum sex-hormone binding globulin levels (r=0.22; P<0.001). CONCLUSIONS: This study shows that the transdermal self-adjusted hormone replacement therapy (HRT) model introduced is suitable for studies on endocrine changes during postmenopausal ERT. The finding of poor correlation between serum E(2) levels and KI emphasizes the importance of hormonal measurements during postmenopausal HRT. PMID- 11888840 TI - The role of sex steroids in the regulation of insulin sensitivity and serum lipid concentrations during male puberty: a prospective study with a P450-aromatase inhibitor. AB - OBJECTIVE: Our purpose was to study the sex steroid-mediated changes in serum insulin and lipid concentrations in boys during puberty. DESIGN AND METHODS: We treated boys with constitutional delay of puberty either with testosterone plus placebo or with testosterone plus an aromatase inhibitor, letrozole, which inhibits the conversion of androgens to oestrogens. We demonstrated previously that during treatment with testosterone plus letrozole the increase in testosterone concentration was more than 5-fold higher than during treatment with testosterone plus placebo. The concentrations of 17beta-oestradiol, IGF-I and IGF binding protein-3 increased during testosterone-plus-placebo treatment, but during testosterone-plus-letrozole treatment the concentrations remained unchanged. These divergent changes in the two groups enabled us to study the effects of sex steroids and GH on insulin sensitivity and lipid concentrations. RESULTS: The insulin concentration in the testosterone-plus-placebo-treated group did not change. In contrast, in the testosterone-plus-letrozole-treated group, the concentration decreased during letrozole treatment, indicating improved insulin sensitivity. Changes in insulin and IGF-I concentrations within 12 and 18 months were correlated. In the testosterone-plus-placebo-treated group, the high density lipoprotein cholesterol concentration did not change but in the testosterone-plus-letrozole-treated group the concentration decreased. The concentrations of low-density lipoprotein cholesterol (LDL-cholesterol) and triglycerides did not change in either of the groups. CONCLUSIONS: The findings indicate that androgens do not directly alter insulin sensitivity in boys during puberty. In contrast, the observations suggest tight regulation of glucose- insulin homeostasis by GH in boys at this stage. Furthermore, our findings indicate that sex steroids do not significantly participate in the regulation of serum concentrations of LDL-cholesterol or triglycerides in boys during early and mid-puberty. PMID- 11888841 TI - Pulsatile secretory characteristics of allopregnanolone, a neuroactive steroid, during the menstrual cycle and in amenorrheic subjects. AB - OBJECTIVE: To investigate whether allopregnanolone, a neuroactive steroid involved in modulating behavioural and neuroendocrine functions, shows episodic secretion in eumenorrheic women, during the follicular and luteal phases of the menstrual cycle, and in women with stress-induced amenorrhea. PATIENTS: Six eumenorrheic women and 14 women with hypothalamic amenorrhea were enrolled for the present study. METHODS: All subjects underwent hormonal evaluation in baseline conditions and a pulsatility study to determine LH, cortisol and allopregnanolone episodic release. Eumenorrheic subjects were investigated twice, in the follicular phase (days 3-7) and in the luteal phase (days 18-22) of the menstrual cycle. LH, FSH, prolactin, estradiol, phosphate, DHEA, allopregnanolone and cortisol levels were evaluated in each case. RESULTS: In healthy women, serum gonadotropin and gonadal steroid levels were significantly lower (P<0.01 and P<0.05 respectively) than those in amenorrheic subjects. Allopregnanolone was higher in amenorrheic subjects and during the luteal phase, compared with the follicular phase, of eumenorrheic subjects (P<0.01). Pulse analysis revealed a significant episodic discharge of allopregnanolone in all subjects (follicular phase 6.5+/-0.3 peaks/6 h and luteal phase 5.5+/-0.4 peaks/6 h, hypothalamic amenorrhea 7.0+/-0.7 peaks/6 h) with higher pulse amplitude in amenorrheic subjects and during the luteal phase compared with the follicular phase of the eumenorrheic subjects (P<0.05). Moreover, the specific concordance index demonstrated that allopregnanolone is coupled with LH only during the luteal phase of the cycle and with cortisol during both phases. Allopregnanolone cortisol coupling was also observed in amenorrheic subjects. CONCLUSIONS: Allopregnanolone is secreted episodically. Both the ovary and adrenal glands release this steroid hormone and it shows temporal coupling with LH only during the luteal phase, with cortisol during both the studied phases of the menstrual cycle in eumenorrheic women and again with cortisol in hypothalamic amenorrheic patients. PMID- 11888842 TI - Spermaturia and serum hormone concentrations at the age of puberty in boys prenatally exposed to polychlorinated biphenyls. AB - OBJECTIVE: To determine whether prenatal exposure to polychlorinated biphenyls (PCBs) with possible hormone-disrupting effects is capable of affecting sexual differentiation in boys at the age of puberty. DESIGN: Following analysis for PCB in their umbilical cords, 196 boys from a Faroese birth cohort were examined for the development of puberty at 14 years of age. METHODS: Physical examination included determination of Tanner stages and testicular size. A morning urine sample was centrifuged and examined for the presence of sperm. Serum was analyzed for sex hormones. RESULTS: twenty boys (10.2%) had abnormalities in testicular development, mainly cryptorchidism. only three of them had a positive spermaturia test, but the level of exposure to pcbs in this group had not been increased. occurrence of spermaturia in 58 of the remaining 176 boys was also not associated with pcb exposure but showed highly significant associations with tanner stages and testicular size. serum concentrations of testosterone, fsh and lh were higher in boys with spermaturia, while sex hormone-binding globulin was lower and no difference occurred in inhibin b. serum hormone parameters showed only weak associations with the level of prenatal pcb exposure. CONCLUSIONS: These findings support the validity of spermaturia as a useful indicator of puberty, although a substantial rate of false negatives must be taken into account. Despite the wide range of exposure to PCB, the findings did not reveal any definite associations with the development of puberty. However, because of the limited size of the cohort, small effects cannot be excluded. PMID- 11888843 TI - Dehydroepiandrosterone in relation to other adrenal hormones during an acute inflammatory stressful disease state compared with chronic inflammatory disease: role of interleukin-6 and tumour necrosis factor. AB - OBJECTIVE: Serum levels of dehydroepiandrosterone (DHEA) and DHEA sulphate (DHEAS) are low in chronic inflammatory diseases, although the reasons are unexplained. Furthermore, the behaviour of serum levels of these hormones during an acute inflammatory stressful disease state is not well known. SUBJECTS AND METHODS: In this study in patients with an acute inflammatory stressful disease state (13 patients undergoing cardiothoracic surgery) and patients with chronic inflammation (61 patients with inflammatory bowel diseases (IBD)) vs. 120 controls, we aimed to investigate adrenal hormone shifts looking at serum levels of DHEA in relation to other adrenal hormones. Furthermore, we tested the predictive role of serum tumour necrosis factor (TNF) and interleukin-6 (IL-6) for a change of serum levels of DHEA in relation to other adrenal hormones. RESULTS: The molar ratio of serum levels of DHEA/androstenedione (ASD) was increased in patients with an acute inflammatory stressful disease state and was decreased in patients with chronic inflammation. The molar ratio of serum levels of DHEAS/DHEA was reduced during an acute inflammatory stressful disease state and was increased in patients with chronic inflammation. A multiple linear regression analysis revealed that elevated serum levels of TNF were associated with a high ratio of serum levels of DHEA/ASD in all groups (for IL-6 in patients with an acute inflammatory stressful disease state only), and, similarly, elevated serum levels of TNF were associated with a high ratio of serum levels of DHEAS/DHEA only in IBD (for IL-6 only in healthy subjects). CONCLUSIONS: This study indicates that changes of serum levels of DHEA in relation to serum levels of other adrenal hormones are completely different in patients with an acute inflammatory stressful disease state compared with patients with chronic inflammation. The decrease of serum levels of DHEAS and DHEA is typical for chronic inflammation and TNF and IL-6 play a predictive role for these changes. PMID- 11888844 TI - Decreased steroidogenic enzyme 17,20-lyase and increased 17-hydroxylase activities in type 2 diabetes mellitus. AB - OBJECTIVE: To analyze activities of adrenal steroidogenic enzymes in type 2 diabetes mellitus, serum levels of 11 steroid hormones were measured simultaneously. SUBJECTS: We studied 130 patients with type 2 diabetes mellitus (74 men and 56 women between the ages of 40 and 69 years), whose blood glucose control had been poor (more than 10% in HbA(1c)). Age-matched normal subjects served as the control group. METHODS: Serum levels of steroid hormones (pregnenolone (Preg), progesterone (Prog), deoxycorticosterone (DOC), corticosterone (B), 17-hydroxypregnenolone (17-OH-Preg), 17-hydroxyprogesterone (17-OHP), 11-deoxycortisol (S), cortisol (F), dehydroepiandrosterone (DHEA) and Delta4-androstenedione (Delta4A)) were measured by HPLC/RIA methods. Fasting plasma glucose (FPG), HbA(1c), ACTH, serum immunoreactive insulin (IRI) and DHEA sulfate (DHEA-S) were also measured. We analyzed product/precursor ratios to assess relative activities of adrenal steroidogenic enzymes. RESULTS: Serum levels of ACTH and F were high and DHEA and DHEA-S were low in both male and female patients under poor blood glucose control. Following 6-months treatment with diet only or with sulfonylurea, FPG and HbA(1c) improved, and blood concentrations of ACTH and F decreased while DHEA and DHEA-S levels increased to within the normal range. DHEA/17-OH-Preg and Delta4A/17-OHP ratios, reflecting 17,20-lyase activity, were low before treatment and recovered to the normal range after treatment, and 17-OH-Preg/Preg and 17-OHP/Prog ratios, reflecting 17 hydroxylase activity, were high before treatment, and fell within the normal range after treatment. 3beta-Hydroxysteroid dehydrogenase, 21-hydroxylase and 11beta-hydroxylase activities remained within the normal range both before and after treatment. CONCLUSIONS: These data suggest that the decrease in DHEA and DHEA-S concentrations together with the high F levels that occur in patients with type 2 diabetes mellitus is associated with low 17,20-lyase and high 17 hydroxylase activity in the adrenal steroidogenic enzymes. High insulin concentrations may further lower DHEA and DHEA-S levels. PMID- 11888845 TI - Survivin: a novel neuroendocrine marker for pheochromocytoma. AB - OBJECTIVE: To study survivin expression in human adrenal medulla and in benign and malignant pheochromocytoma tissue as a tool to predict tumor metastatic potential and prognosis. DESIGN: Blinded study to assess the role of the anti survivin antibody in chromaffin cells. METHODS: We performed immunohistochemistry with a purified rabbit-polyclonal anti-survivin antibody on 39 formalin-fixed and paraffin-embedded pheochromocytoma/paraganglioma specimens, and on 10 normal adrenal medulla samples from patients unaffected by a chromaffin cell tumor. Fourteen samples were from 14 patients with benign pheochromocytoma (<8 year follow-up, mean 5.2 years), 18 specimens were from 12 patients with malignant pheochromocytoma (<13 year follow-up, mean 6.3 years), 5 samples were from 2 patients with malignant paraganglioma (<6 year follow-up, mean 4 years), and 2 specimens from 2 patients with benign paraganglioma (<7 year follow-up, mean 5.5 years). Malignancy was defined by metastases in non-chromaffin tissues. Staining intensity with the anti-survivin antibody was scored from 0 (none) to 3+ (heavy). Tissues from human kidney, breast, and melanoma served as controls. RESULTS: All pheochromocytoma/paraganglioma specimens stained either 2+ or 3+. By analysis of variance (ANOVA), there was no statistically significant difference between the staining intensity of benign and malignant samples. All normal adrenal medulla specimens stained positively with anti-survivin but to a lesser degree than the chromaffin cell tumors (P<0.01). CONCLUSIONS: Based on these findings, we conclude that (i) survivin may represent a novel neuroendocrine marker for chromaffin cell tumors, and (ii) survivin does not appear to reliably distinguish benign from malignant pheochromocytomas/paragangliomas and thus does not identify patients at risk of recurrent disease. PMID- 11888846 TI - Raised serum levels of interleukin-8 and interleukin-18 in relation to bone metabolism in endogenous Cushing's syndrome. AB - OBJECTIVE: It is well known that patients with endogenous Cushing's syndrome (CS) have decreased bone mass and enhanced risk for osteoporotic fractures, secondary to decreased bone formation and increased bone resorption. Immunological mediators, such as cytokines, have recently been shown to influence bone metabolism, and in the present study we examined serum levels of several cytokines, with known or potential effects on bone homeostasis, in 33 consecutive recruited untreated CS patients and 33 age-, sex- and body mass index-matched healthy controls. METHODS: Cytokine levels were measured by enzyme immunoassay and bone mass by dual-energy X-ray absorptiometry. RESULTS: Our main findings were (i) interleukin (IL)-8 and IL-18 levels were significantly increased in CS patients compared with controls. (ii) Levels of both IL-8 and IL-18 were positively correlated to serum cortisol. (iii) For serum levels of the 'classical' resorptive cytokines, i.e. IL-6 and tumor necrosis factor alpha, no significant differences were found between CS patients and controls. (iv) Raised IL-18 levels were correlated with decreased osteocalcin levels in CS patients. CONCLUSIONS: Our results demonstrated that CS patients have markedly elevated levels of the proinflammatory cytokines IL-8 and IL-18 in spite of high levels of the immunosuppressive hormone cortisol. These cytokines may be involved in the pathogenesis of disturbed bone homeostasis in CS. PMID- 11888847 TI - Abnormal release of incretins and cortisol after oral glucose in subjects with insulin-resistant myotonic dystrophy. AB - OBJECTIVE: Although the incretins, gastric inhibitory polypeptide (GIP) and glucagon-like peptide-1 (GLP-1), as well as glucagon and cortisol, are known to influence islet function, the role of these hormones in conditions of insulin resistance and development of type 2 diabetes is unknown. An interesting model for the study of hormonal perturbations accompanying marked insulin resistance without concomitant diabetes is myotonic dystrophy (DM1). DESIGN: The work was carried out in an out-patient setting. METHODS: An oral glucose tolerance test was performed in 18 males with DM1 and 18 controls to examine the release of incretins and counter-regulatory hormones. Genetic analyses were also performed in patients. RESULTS: We found that the increment in GLP-1 after oral glucose was significantly greater in patients, while there was no significant difference in GIP or glucagon responses between patients and controls, although long CTG repeat expansions were associated with a more pronounced GIP response. Interestingly, the GLP-1 response to oral glucose correlated with the insulin response in patients but not in controls whereas, in controls, the insulin response closely correlated with the GIP response. Furthermore, cortisol and ACTH levels increased paradoxically in patients after glucose; this was more pronounced in patients with long CTG repeat expansions. CONCLUSIONS: This study showed that the GLP-1 and ACTH/cortisol responses to oral glucose are abnormal in insulin-resistant DM1 patients and that CTG triplet repeats are linked to GIP release. These abnormalities may contribute both to the severe insulin resistance and hyperinsulinemia in DM1 and to the preservation of adequate islet function, enabling glucose tolerance to be normal in spite of this marked insulin resistance in DM1. PMID- 11888848 TI - Effects of acute and chronic hypercalcemia on parathyroid function and circulating parathyroid hormone molecular forms. PMID- 11888849 TI - The influence of acute and chronic hypercalcemia on the parathyroid hormone response to hypocalcemia in rabbits. AB - OBJECTIVE: To investigate the influence of acute and chronic hypercalcemia on the parathyroid hormone (PTH) response to hypocalcemia. DESIGN: The PTH response to hypocalcemia has been evaluated in three groups of rabbits: Group I, normal rabbits, Group II, normal rabbits subjected to an acute hypercalcemic clamp (induced by CaCl(2) infusion) and Group III, rabbits with chronic hypercalcemia (due to surgical reduction of renal mass). RESULTS: In Group I (baseline Ca(2+)=1.69+/-0.02 mM), hypocalcemia resulted in stimulation of PTH secretion which reached a maximum (PTHmax) of 91.7+/-6.4 pg/ml. In rabbits from Group II, which also had normal baseline Ca(2+) (1.70+/-0.02 mM), plasma Ca(2+) was maintained at an elevated level for 2 h, at around 2.05 mM. The PTH response to hypocalcemia in Group II was attenuated and the PTHmax in these rabbits was 45.6+/-7.4 pg/ml. In rabbits from Group III, baseline Ca(2+) was elevated (2.06+/ 0.06 mM) for 1 month. The PTH response to hypocalcemia in Group III was esentially the same as in Group I and PTHmax reached levels of 94.8+/-9.9 pg/ml. CONCLUSIONS: A difference in PTH response to hypocalcemia has been found in rabbits after exposure to either acute or chronic hypercalcemia. After acute hypercalcemia, an attenuated PTH response to hypocalcemia has been identified. Chronic hypercalcemia, however, did not influence the PTH response to hypocalcemia. PMID- 11888850 TI - Dexamethasone impairs insulin signalling and glucose transport by depletion of insulin receptor substrate-1, phosphatidylinositol 3-kinase and protein kinase B in primary cultured rat adipocytes. AB - OBJECTIVE: Glucocorticoid excess leads to insulin resistance. This study explores the effects of glucocorticoids on the glucose transport system and insulin signalling in rat adipocytes. The interaction between glucocorticoids and high levels of insulin and glucose is also addressed. DESIGN AND METHODS: Isolated rat adipocytes were cultured for 24 h at different glucose concentrations (5 and 15 mmol/l) with or without the glucocorticoid analogue dexamethasone (0.3 micromol/l) and insulin (10(4) microU/ml). After the culture period, the cells were washed and then basal and insulin-stimulated glucose uptake, insulin binding and lipolysis as well as cellular content of insulin signalling proteins (insulin receptor substrate-1 (IRS-1), IRS-2, phosphatidylinositol 3-kinase (PI3-K) and protein kinase B (PKB)) and glucose transporter isoform GLUT4 were measured. RESULTS: Dexamethasone in the medium markedly decreased both basal and insulin stimulated glucose uptake at both 5 and 15 mmol/l glucose (by approximately 40 50%, P<0.001 and P<0.05 respectively). Combined long-term treatment with insulin and dexamethasone exerted additive effects in decreasing basal, and to a lesser extent insulin-stimulated, glucose uptake capacity (P<0.05) compared with dexamethasone alone, but this was seen only at high glucose (15 mmol/l). Insulin binding was decreased (by approximately 40%, P<0.05) in dexamethasone-treated cells independently of surrounding glucose concentration. Following dexamethasone treatment a approximately 75% decrease (P<0.001) in IRS-1 expression and an increase in IRS-2 (by approximately 150%, P<0.001) was shown. Dexamethasone also induced a subtle decrease in PI3-K (by approximately 20%, P<0.01) and a substantial decrease in PKB content (by approximately 45%, P<0.001). Insulin stimulated PKB phosphorylation was decreased (by approximately 40%, P<0.01) in dexamethasone-treated cells. Dexamethasone did not alter the amount of total cellular membrane-associated GLUT4 protein. The effects of dexamethasone per se on glucose transport and insulin signalling proteins were mainly unaffected by the surrounding glucose and insulin levels. Dexamethasone increased the basal lipolytic rate (approximately 4-fold, P<0.05), but did not alter the antilipolytic effect of insulin. CONCLUSIONS: These results suggest that glucocorticoids, independently of the surrounding glucose and insulin concentration, impair glucose transport capacity in fat cells. This is not due to alterations in GLUT4 abundance. Instead dexamethasone-induced insulin resistance may be mediated via reduced cellular content of IRS-1 and PKB accompanied by a parallel reduction in insulin-stimulated activation of PKB. PMID- 11888851 TI - Parathyroid hormone induces formation of new cancellous bone with substantial mechanical strength at a site where it had disappeared in old rats. AB - OBJECTIVE: The present study addresses the question--can PTH induce formation of trabeculae in areas where cancellous bone has disappeared? Two-year-old male rats were chosen, because in this aged animal model the distal femurs have almost no cancellous bone, and the marrow cavity has reached a substantial dimension. DESIGN: The rats were injected for 56 days with either PTH(1-34), 15 nmol/kg/day (62.5 microg/kg/day), or vehicle. METHODS: Transverse specimens, 2-mm high, were cut from the distal femoral metaphysis. Marrow cavity diameters and cancellous bone trabeculae were analysed by a micro-computerized tomography scanner. The cancellous bone within the cortical and endocortical rim of each specimen was submitted to a biomechanical compression test. Furthermore, the cancellous bone was studied by dynamic tetracycline labelling and histomorphometry. RESULTS: In the vehicle-injected group the trabecular bone volume was 0% (0-1.4), median (range). All PTH-injected rats had trabeculae in the distal metaphysis and the trabecular bone volume (6.7% (2.3-12.0)) was markedly increased (P<0.003). The median trabecular thickness was increased (P<0.003) in the PTH-injected rats (118 microm (104-125)) compared with the vehicle group (0 microm (0-71)). The compressive stress was increased (P<0.003) in the PTH-injected group (0.7 MPa (0.1-2.1)) compared with the vehicle-injected group (0 MPa (0-0.4)). The histomorphometry revealed that only 3 animals of the 10 in the vehicle-injected group had trabeculae in the distal femoral metaphysis. All PTH-injected animals (12 of 12) had continuous trabecular bone network in the marrow cavity. CONCLUSION: Intermittent PTH treatment induced marked formation of new cancellous bone trabeculae with substantial mechanical strength, at a site where it had disappeared in old rats. PMID- 11888852 TI - Evidence of angiotensin II involvement in prolactin secretion in response to hemorrhage in adrenodemedullated and guanethidine-treated rats. AB - OBJECTIVE: The present experiments were designed to investigate the influence of the renin--angiotensin system (RAS) on prolactin secretion in response to hemorrhage (1.2 ml/100 g body weight (bw)/2 min). METHODS AND RESULTS: Male Wistar rats (250-300 g) were divided into the following experimental groups. (i) Sham-operated animals submitted to intravenous administration of [Sar(1),Thr(8)] angiotensin II (sarthran), an angiotensin II antagonist (750 ng/100 g bw as a bolus plus an infusion of 25 ng/100 g bw/min over 30 min), which did not alter the prolactin secretion in response to hemorrhage. (ii) Animals submitted to adrenodemedullation which by itself increased the prolactin secretion in response to hemorrhage by 274% (P<0.01). However, sarthran infusion into adrenodemedullated rats completely blocked this increased prolactin secretion in response to hemorrhage (P<0.01). (iii) Intact animals submitted to blockade of sympathetic noradrenergic pathways by pretreatment with guanethidine (10 mg/100 g bw), which also increased the prolactin secretion in response to hemorrhage by 55% (P<0.01). This increased prolactin secretion in response to hemorrhage observed in guanethidine-treated rats was completely blocked by sarthran preinfusion (P<0.01). (iv) Adrenodemedullated animals pretreated with guanethidine, which abolished the prolactin secretion induced by hemorrhage. CONCLUSIONS: Our data suggest a role for circulating catecholamines in the prolactin secretion response to stress. In addition, the experiments reported here demonstrate that RAS has a stimulatory effect on prolactin secretion in circumstances in which sympathetic activity or adrenomedullary secretion is suppressed. These are the first data demonstrating that a physiological prolactin secretion response to stress depends on the RAS. PMID- 11888853 TI - Down-regulation of proliferation and up-regulation of apoptosis by gonadotropin releasing hormone agonist in cultured uterine leiomyoma cells. AB - OBJECTIVE: To elucidate the direct effects of gonadotropin-releasing hormone agonist (GnRHa) on the growth of human uterine leiomyoma cells, cell proliferation and apoptosis in cultured leiomyoma cells treated with GnRHa were investigated. METHODS: Isolated leiomyoma cells were subcultured in DMEM supplemented with 10% FBS for 5 days and stepped down to serum-free conditions for an additional 6 days in the presence or absence of graded concentrations of GnRHa (10(-9) mol/l to 10(-12) mol/l). The effects of GnRHa on the number of viable cells, expression of proliferating cell nuclear antigen (PCNA), Fas and Fas ligand, and apoptosis in cultured leiomyoma cells were examined by MTT (3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide) assay, immunocytochemical analysis, Western blot analysis and TUNEL assay respectively. RT-PCR was performed to detect the expression of GnRH receptor mRNA in cultured leiomyoma cells. RESULTS: Treatment with GnRHa resulted in a decrease in the number of cultured viable leiomyoma cells assessed by MTT assay in a dose dependent manner compared with that in control cultures (P<0.01). The growth inhibition of cultured leiomyoma cells treated with GnRHa in concentrations higher than 10(-10) mol/l was associated with the suppression of the proliferative potential characterized by a decrease in PCNA-positive rate of the cultured cells (P<0.01) and an increase in the apoptosis-positive rate assessed by TUNEL assay (P<0.05 and P<0.01). GnRHa markedly increased the expression of Fas and induced the expression of Fas ligand in the cultured leiomyoma cells on the basis of Western blot analysis. These direct effects of GnRHa on the number of viable cultured leiomyoma cells, PCNA-positive rate, apoptosis-positive rate and Fas/Fas ligand expression in the cultured leiomyoma cells were only attained after the 4-day treatment. RT-PCR analysis revealed that GnRH receptor mRNA was expressed in cultured leiomyoma cells. CONCLUSIONS: The present results demonstrate that GnRHa directly inhibits the growth of human uterine leiomyoma cells by suppressing cell proliferation and inducing apoptosis, which might be associated with the increase in Fas expression and the induction of Fas ligand expression in the cells. PMID- 11888855 TI - [Cytogenetics, cytogenomics and cancer]. AB - Chromosomal study in malignancy has demonstrated the pivotal role of somatic chromosomal rearrangements in oncogenesis and tumoral progression. Structural or quantitative these abnormalities can now be studied in great details with the various Fish techniques, including CGH on chromosomes or in a near future on micro arrays. The multistep pattern of most solid tumors is characterized and their genomic abnormalities more and more used for the diagnosis and the prognosis. PMID- 11888854 TI - [What hysterectomy procedure should be carried out for cancer of the endometrium in stage I-II]. PMID- 11888856 TI - [From cytogenetics to cytogenomics of bladder cancers]. AB - Bladder cancers are classified as: transitional cell carcinoma (TCC), the most frequent in Europe/USA, squamous cell carcinoma (SCC), more frequent in the Middle East and in Africa, adenocarcinoma and small cell carcinoma, rare. TCC exhibit pseudo diploid karyotypes with only a few anomalies in early stages, evolving towards pseudo-tetraploides complexes karyotypes. Partial or complete monosomy 9 (-9) is an early event, found in half cases. Deletion (11p) or -11 is found in 20-50% of cases, more often in high grade and invasive tumours. Del(13q) is found in 25% of cases and correlated with high grade/stage; tumours with Rb alterations are invasives. Del(17p) is a late event, found in 40% of cases; P53 alterations are correlated with grade and stage, tumour progression, and a worse prognosis. Del(1p), i(5q), +7, and many other rearrangements - more often deletions than duplications - are frequently found. These losses of heterozygocity point to a multistep complex process involving tumor suppressor genes. In SCC, monosomy 9 is also an early event, even more frequent than in TCC; homozygous deletion of P16 is frequent. Trisomy 7 seems to be more frequent than in TCC. Chromosome 17 is often implicated, especially in high grades/stages; the profile of mutations of P53 is different from what is found in TCC. Allelic losses of 3p, 8p, 9p, 9q, 17p are frequent. The karyotype is more complex in advanced grades/stages, as in TCC. PMID- 11888857 TI - [From "monocolor" karyotype to "multicolor" karyotype: applications of M-Fish in hematology and oncology]. AB - Since the establishment of human karyotype in 1956, human cytogenetic has quickly progressed. The description of the Philadelphia chromosome in 1960 led up to new applications of cytogenetic in the fields of hematology and oncology. The initial techniques allowed only uniform staining of chromosomes, limiting the detection of most structural rearrangements. Many approaches aimed to gain a better knowledge of chromosomal structure, a better understanding of rearrangements, and a better identification of the chromosomes were developed: autoradiography, banding techniques, electronic microscopy. Since 1980, new developments in clinical cytogenetic and molecular biology have occurred. In situ labeling using non-radioactive probes onto chromosomes and nuclei was developed: fluorescence in situ hybridization (Fish) was born. Fish allows detecting many chromosomal abnormalities of number and/or structure. The major limitation of this technique is that its use should be based on known indications for the choice of the probe. Multicolor karyotype (M-Fish or Sky), the most recent development of Fish on metaphase spreads, allows to overcome this limit. As shown here in three examples, M-Fish allows to describe precisely complex rearrangements in hematological malignancies and solid tumors. Finally, if no metaphase is available, comparative genomic hybridization (CGH) can be performed to detect and simultaneously localize on chromosomes gains or losses in genomic DNA. PMID- 11888858 TI - [Update in radiation-induced neoplasms: genetic studies]. AB - Radiation induced tumors are a possible (very) late complications of radiotherapy. The evaluation of the risks of radiation-induced tumors has been presented in different epidemiological studies, with the evaluation of the relative risk for different tissues. But, the genetic studies are rare, and no global theory exists. Two cytogenetic profiles are described, one with translocations and one with genetic material losses, evoking two different genetic evolutions. Two questions are stated. What are the radiation-induced genetic mechanisms? Is it possible to differentiate the radiation-induced and spontaneous tumors with genetic approaches? With 37 cytogenetic cases, 12 analyzed in our laboratory, the radiation-induced tumors were characterized by genetic material losses. An anti-oncogenic evolution is probable. A new molecularly study confirm these results. Only thyroid tumors do not have this evolution. For tumors with simple karyotype, like meningioma, radiation-induced tumors seem to be more complex than spontaneous tumors. But for the others, the differentiation is impossible to be done with cytogenetic. The mechanism of the chromosomic material losses in unknown, but some hypothesis are discussed. PMID- 11888859 TI - [Cancer networks]. AB - The complexity of legislation and the obstacle race run by all promoters in search of approval and funding for their projects could have greatly hindered the development of healthcare networks in France, or at least discouraged many promoter. Yet social and health professionals, along with certain healthcare service staff, considered these networks highly useful for decompartmentalising the French healthcare system. They would favour patient-oriented projects. It is not a surprise that the number of networks in the field of oncology is expanding rapidly, with more than 60 projects registered by public authorities by the end of 1999. Cancers are chronic diseases which often bring into play the patient's vital prognosis, as well as psychological and social consequences - sometimes dreadful - for patients and their relatives. Cancer patient management calls for a multidisciplinary approach in elaborating the therapeutic project and providing psychological support. Continuity and consistency of care require a partnership between different care providers from different specialities, with different medical practice and environments - hospital or home care, public or private practice. Though so much information has become available in the medical literature, quality of care demands that each healthcare professional keeps fully aware of the latest diagnostic or therapeutic findings and techniques in his/her field. Certain cancer networks have become a place where care providers and establishments can share methods and medical practice. Network promoters and members have thus been able to regain control over the functioning and the future of their profession. Patients have also obtained a place in decisions concerning their own health. Setting up healthcare networks will certainly shake up the tranquil system which the WHO has reported as the world's best health care system at the dawn of this century. However, this will also probably contribute to maintaining this excellence. PMID- 11888860 TI - [Standards, Options and Recommendations for non metastatic breast cancer patients]. AB - CONTEXT: The "Standards, Options and Recommendations" (SOR) project, started in 1993, is a collaboration between the Federation of French Cancer Centers (FNCLCC), the 20 French Cancer Centers, and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and the outcome of cancer patients. The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts, with feedback from specialists in cancer care delivery. OBJECTIVES: To develop clinical practice guidelines for non metastatic breast cancer patients according to the definitions of the Standards, Options and Recommendations project. METHODS: Data were identified by searching Medline , web sites, and using the personal reference lists of members of the expert groups. Once the guidelines were defined, the document was submitted for review to 148 independent reviewers. RESULTS: This article is an update of the version published in 1996. The modified 2001 version of the standards, options and recommendations takes into account new information published. Important changes have been made in terms of clinical practice. They concern loco-regional and general therapy. Regarding loco-regional treatment, a increased dose to the tumor bed should be needs to be systematically delivered to the tumor bed for women under 50 years. The analysis of margin involvement is essential and constitutes an important factor for therapeutic decision. Regarding general therapy, hormone therapy with tamoxifen appeared, in 2001, as one of the standards in non-menopausal and oestrogen receptor-positive patients. Concerning patients with N- tumors, the standards and therapeutic options were refined with regard to the notion of metastatic relapse risk. Chemotherapy constitutes one of the standard treatments for non-menopausal women with one or more factor(s) predictive of metastatic relapse. PMID- 11888861 TI - [Involvement of FAK, PI3-K and PKC in cell adhesion induced by microtubule disruption]. AB - We have previously shown that microtubule disruption results in an increase in cell adhesion to ECM proteins. In this work we show that this enhanced cell attachment was completely abolished by specific inhibitors of tyrosine-kinases, PI3-K and PKCs. Microtubule depolymerisation was associated with an important increased in tyrosine phosphorylation of FAK and paxilline, as well as with subcellular localisation of PKCgamma, delta and epsilon. We also observed significant alterations in actin cytoskeleton leading to reduced cell spreading. Thus, microtubule depolymerisation appears to activate various intracellular kinases that lead to actin cytoskeletal changes and to an increase of integrin dependent adhesion. Whether this enhanced attachment is due to intracellular events resulting in changes in integrin affinity or avidity remains to be determined. PMID- 11888862 TI - [Antecedents of cytological screening among patients treated for invasive cervical neoplasm]. AB - OBJECTIVE: The aim of this study is to evaluate if the seriousness of invasive neoplasms of the uterin cervix actually observed is related to the apparition of rapid onset cases. POPULATION: 219 invasive cancers of the cervix treated from 1988 to 1999 in a gynecological oncologic department. METHODS: prognostic factors of cancers have been studied and compared to those of cases treated between 1975 and 1980 in the same department. The existence of cytological screenings has been searched and results have been analysed. RESULTS: Cervical cancers treated during the last 12 years have more serious prognostic factors than those treated during the former period. This evolution is shown by a progression of advanced stages of + 10.2%, an increase of lymph node invasion in proximal stages of + 13,4% and the doubling of number of adenocarcinomas. Though no change of the natural history of cancers has been proved. Only 42% of patients had profited of a cervical screening and cancers diagnosed at "stage I" are statistically more numerous in this group. Improvement should be brought up in the quality of samplings and of the care of abnormal results of cytological screening. PMID- 11888863 TI - [Public health: a true concern in the French political scene]. AB - A recent report of the Senate revealed the difficulties of elaborating an effective cancer policy in France. Senators particularly pointed to the necessity of establishing cancer care priorities and to the tremendous need for medical coordination. This paper will develop the annual state debate on social security funding. The authors will show that health policy is entirely dependent on health economics. We believe that health priorities and health outcomes should be defined before any financial decision is made, this preliminary debate being an essential contribution to evidence-based policy-making in public health. PMID- 11888864 TI - Predicting survival in melanoma. PMID- 11888865 TI - Fighting wars, winning battles. PMID- 11888866 TI - Can specific axillary radiotherapy be omitted in undissected, clinically node negative patients who undergo breast-conserving therapy? PMID- 11888867 TI - Serum TA90 immune complex assay can predict outcome after resection of thick (> or =4 mm) primary melanoma and sentinel lymphadenectomy. AB - BACKGROUND: We hypothesized that the postoperative serum level of TA90-IC, an immune complex of a 90-kDa tumor-associated antigen and its antibody, might have a significant correlation with recurrence and survival in patients with thick primary melanomas. METHODS: We used our prospective melanoma database to identify all patients who underwent wide local excision and sentinel lymphadenectomy for primary melanomas > or =4 mm and from whom sera had been collected and cryopreserved within 6 months after surgery. These sera were analyzed in a blinded fashion for TA90-IC status by using our double-determinant enzyme-linked immunosorbent assay. Results were correlated with disease-free survival (DFS) and overall survival (OS). Standard prognostic factors for melanoma were then compared with TA90-IC status for the prediction of DFS and OS. RESULTS: The sensitivity and specificity of the TA90-IC assay for predicting recurrence were 70% and 85%, respectively. Five-year DFS and OS rates were higher for the TA90-IC negative group than the positive group. The differences in DFS and OS between the TA90-IC-negative and -positive groups were significant. At a median follow-up of 25 months, multivariate analysis identified postoperative TA90-IC status and sex as significant predictors of DFS. TA90-IC status was the only independent prognostic factor with multivariate analysis. CONCLUSIONS: TA90-IC status after resection of thick primary melanoma accurately predicts outcome. A positive postoperative TA90-IC level might affect a decision regarding adjuvant therapy, regardless of regional nodal status. PMID- 11888868 TI - Prognostic factors after isolated limb infusion with cytotoxic agents for melanoma. AB - BACKGROUND: Isolated limb perfusion (ILP) with cytotoxic agents is a remarkably effective but complex technique used to treat locally recurrent and metastatic melanoma confined to a limb. Isolated limb infusion (ILI), essentially a low-flow ILP performed without oxygenation via percutaneous catheters, has been developed as a simpler alternative. METHODS: The outcome in 135 patients treated by ILI was reviewed. RESULTS: The overall response rate in the treated limb was 85% (complete response [CR] rate 41%, partial response rate 44%). Median response duration response was 16 months (24 months for patients with CR). Median patient survival was 34 months. In those with a CR, the median survival was 42 months. CR rate and survival time decreased with increasing disease stage. Patients aged >70 years had a better overall response than younger patients. On multivariate analysis, factors associated with an improved outcome were a lower stage of disease, a final limb temperature >37.8 degrees C, and a tourniquet time >40 minutes. CONCLUSIONS: The frequency and duration of responses after ILI were comparable to those achieved by conventional ILP. The ILI technique is particularly useful for older patients who might not be considered suitable for conventional ILP. PMID- 11888869 TI - Frequency of nonsentinel lymph node metastasis in melanoma. AB - BACKGROUND: Completion lymph node dissection (CLND) may not be necessary for some patients because nodal metastasis is rarely detected beyond the sentinel lymph nodes (SLNs). This analysis was performed to determine, among patients with positive SLNs, the rate of nodal metastasis found in nonsentinel nodes (NSNs). METHODS: This analysis includes patients with positive sentinel nodes, detected by hematoxylin and eosin (H&E) staining or immunohistochemistry (IHC), who then underwent CLND. RESULTS: This analysis included 274 patients with at least one positive SLN who underwent CLND of 282 involved regional nodal basins. Of the 282 SLN-positive nodal basins, 45 (16%) were found to have positive NSNs in the CLND specimen. Breslow thickness, Clark level, presence of ulceration, histological subtype, presence of vertical growth phase, evidence of regression, presence of lymphovascular invasion, number of positive SLNs, age, sex, and presence of multiple draining nodal basins were not predictive of positive nodes in the CLND specimen. Patients with SLN metastases detected only by IHC had an equal likelihood of having positive NSNs as those patients with positive SLNs on H&E examination. CONCLUSIONS: No patient population could be identified with minimal risk of non-SLN metastasis. When a positive SLN is identified on either H&E staining or IHC, CLND should be performed routinely. PMID- 11888870 TI - The illusion of the learning phase for lymphatic mapping. AB - BACKGROUND: We provide a statistical analysis of the learning phase for sentinel node biopsy. METHODS: Four learning phases were analyzed: 25, 50, 75, and 150 procedures with a corresponding number of 10, 20, 30, and 60 tumor-positive cases. Critical values of nonidentification rate and false-negative rate were defined. The binomial distribution was used to calculate the probabilities of correctly or incorrectly accepting the quality of the performance, given a certain long-term nonidentification or false-negative rate. RESULTS: The chance of incorrectly reaching a favorable false-negative rate of <10% (critical value) in 20 metastasized patients was 18% for a surgeon with a long-term probability of false-negative procedures of 15%. This chance was reduced to 10% with a learning phase of 60 tumor-positive cases. When this chance has to be further reduced to 5%, the critical value has to be lower in smaller groups of patients: 5% false negative rate in 20 tumor-positive procedures. CONCLUSIONS: A learning phase of at least 150 procedures with 60 tumor-positive cases is needed to draw any reliable conclusion about the quality of sentinel node biopsy. In general, a compromise has to be made between the reliability of the results and the practically achievable number of procedures. PMID- 11888872 TI - Axillary radiotherapy instead of axillary dissection: a randomized trial. Italian Oncological Senology Group. AB - BACKGROUND: Surgical dissection of the axilla is a standard part of the treatment of breast cancer but, by itself, does not improve prognosis; furthermore, most patients with small-sized breast cancer and a clinically uninvolved axilla never develop axillary metastases. We evaluated disease-free and overall survival in patients with early breast cancer treated by breast-conservation surgery without dissection of axillary lymph nodes, receiving or not receiving axillary radiotherapy (RT). METHODS: From 1995 to 1998, 435 patients older than 45 years with breast cancer up to 1.2 cm were randomized, 214 to breast conservation without axillary treatment and 221 to breast conservation plus axillary RT. RESULTS: After a follow-up of 28 to 68 months (median, 42 months), two women (1%) in the no axillary treatment group and one (.5%) in the axillary RT group developed axillary metastases. Rates of distant metastases and local treatment failure were also very low, and 5-year overall survival was 99%. CONCLUSIONS: After a mean of 46 months of follow-up, our results indicate that axillary dissection can be safely avoided in patients with very small invasive carcinomas and a clinically negative axilla. Whether axillary RT should be added can be assessed only by longer follow-up. PMID- 11888871 TI - Systemic irinotecan and regional floxuridine after hepatic cytoreduction in 185 patients with unresectable colorectal cancer metastases. AB - BACKGROUND: This study evaluated our 7-year experience treating unresectable colorectal cancer (CRC) hepatic metastases refractory to systemic 5-fluorouracil. METHODS: A total of 185 patients with unresectable 5-fluorouracil-resistant CRC hepatic metastases underwent surgical cytoreduction. Postoperatively patients received either hepatic arterial floxuridine (FUDR) and systemic irinotecan as part of a phase II trial or no further treatment. RESULTS: Of the 185 patients undergoing surgical cytoreduction, 71 patients received adjuvant irinotecan/FUDR. There were no appreciable differences in synchronous or metachronous lesions or the median number or size of lesions between treatment groups. At a median follow up of 20 months, there were fewer recurrences in patients treated with postoperative irinotecan/FUDR compared with untreated patients for both hepatic and extrahepatic recurrences. Progression-free and overall survival were longer for patients who received irinotecan/FUDR compared with patients who did not receive adjuvant therapy. The 2-year survival rate was significantly better for patients receiving adjuvant therapy compared with patients receiving no additional treatment. Predictors of improved survival included a preoperative carcinoembryonic antigen level <100 ng/dl, >30% postoperative reduction in carcinoembryonic antigen level, and adjuvant therapy. CONCLUSIONS: Combined therapy with irinotecan/FUDR may improve the results of surgical cytoreduction for unresectable CRC hepatic metastases. PMID- 11888873 TI - Pure mucinous carcinoma of the breast: is axillary staging necessary? AB - BACKGROUND: Mucinous carcinoma of the breast (MCB) may be associated with a low risk of axillary metastases. METHODS: To evaluate the incidence of axillary nodal metastasis in MCB, a review of all cases from January 1990 to July 2000 was performed. Pure MCB was defined as all tumor cells being completely surrounded by mucin. Patient demographics, tumor size, estrogen receptor status, total number of dissected lymph nodes, and incidence of nodal metastasis were studied. Deeper sections on the lymph nodes from the pure tumors were performed and stained with low-molecular cytokeratin. RESULTS: Nineteen cases of pure MCB and 41 cases of mixed MCB were identified. Patients with pure MCB were older than those with mixed MCB. Tumor size and estrogen receptor status showed no statistically significant differences between the two groups. None of the patients with pure MCB demonstrated lymph node metastases, whereas 12 of 41 cases with mixed MCB demonstrated metastatic lymph node involvement. CONCLUSIONS: Because pure MCB seems unlikely to metastasize, axillary lymph node staging in these patients may not be necessary. The presence of lymph node metastases strongly indicates the presence of a mixed MCB. PMID- 11888874 TI - Analysis of nipple/areolar involvement with mastectomy: can the areola be preserved? AB - BACKGROUND: Skin-sparing mastectomy (SSM), which involves the resection of the nipple/areolar complex with the breast parenchyma, improves the aesthetic outcome for breast cancer patients. Most patients undergoing SSM desire reconstruction of the nipple/areolar complex for symmetry. These data explore the possibility of preserving the areola in selected mastectomy patients. METHODS: A retrospective analysis of 217 mastectomy patients was conducted to determine the frequency of malignant nipple and/or areola involvement. The association between nipple and/or areola involvement and prognostic factors, including tumor size, stage, nuclear grade, axillary nodal status, and tumor location, was evaluated. RESULTS: The overall frequency of malignant nipple involvement was 23 of 217 (10.6%). In a subgroup of patients with tumors <2 cm, peripheral tumors, and with two positive nodes or less, the incidence of nipple involvement was 6.7%. When the nipple and areolar involvement were analyzed separately, only 2 of 217 patients had involvement of the areola (0.9%). All patients with areolar involvement had stage 3 breast cancer and were located centrally in the breast. CONCLUSIONS: We conclude from these data that nipple preservation is not a reasonable option for mastectomy patients. However, preservation of the areola with mastectomy in selected patients warrants further study. PMID- 11888875 TI - Subareolar and peritumoral injection identify similar sentinel nodes for breast cancer. AB - BACKGROUND: Sentinel lymph node (SLN) mapping with radioisotope and blue dye is rapidly becoming the standard of care for breast cancer. The optimal location for injection of radioisotope and blue dye is still being investigated. The goal of this study was to determine whether blue dye injection into the subareolar (SA) location localized the same sentinel nodes as the peritumoral (PT) location for patients with breast cancer. METHODS: Three hundred thirty-two patients with biopsy-proven operable breast cancer or ductal carcinoma in situ at two institutions underwent SLN mapping. Eighty-three patients had PT injection of blue dye (group 1), and 249 patients had SA injection of blue dye (group 2). All patients underwent PT injection of (99m)Tc-labeled sulfur colloid. RESULTS: The two groups were similar in age, previous biopsy type, and tumor size, location, and histology. The mean number of SLNs identified was 2.4 (range, 0-9) in group 1 and 2.5 (range, 0-11) in group 2. The SLN identification rate was 95% for group 1 and 97% for group 2. The isotope success rate was 94% for both groups. The blue dye success rate was 84% for group 1 and 90% for group 2. The isotope/blue dye concordance rate was 87% for group 1 and 90% for group 2. At a median follow-up of 28 months (range, 14 to 40), there were no axillary recurrences in any of the 332 patients. CONCLUSIONS: These data suggest that delivery of mapping reagents in the SA and PT locations identifies similar lymph nodes. Because of simplicity and the similarity in node identification between SA and PT injection, further investigation of the SA site for delivery of SLN mapping reagents for breast cancer is warranted. PMID- 11888876 TI - The Mayo Clinic experience with multimodality treatment of locally advanced or recurrent colon cancer. AB - BACKGROUND: Patients with incompletely resected locally advanced and recurrent colon cancers have a dismal prognosis. Since 1981, 100 colon cancer patients have been treated with combination therapy including surgical resection, chemotherapy, and external plus intraoperative radiotherapy. METHODS: A prospective computerized intraoperative radiation database identified patients for this retrospective review. Data collection included patient demographics, tumor and treatment variables, and morbidity, recurrence, and survival statistics. RESULTS: The mean age was 55.2 years. Follow-up was available for all patients. Fifty-nine patients have died. Median follow-up of survivors was 70.5 months. Twenty-five patients with locally advanced colon cancer had a median survival of 38.2 months and a 5-year survival of 49%. Eleven of these patients are still free of disease. Seventy-three patients treated for recurrent colon carcinoma had a median survival of 33.3 months from the time of recurrence, with a 5-year survival of 24.7%. Twenty-one are alive without evidence of recurrence. The 38 patients with recurrent disease whose disease was completely resected had a 37.4% 5-year survival. CONCLUSIONS: A multimodality approach using en-bloc surgical resection with radiotherapy and chemotherapy affords some patients with locally advanced and recurrent colon cancer a chance for long-term survival. PMID- 11888877 TI - Safety monitoring of the coliseum technique for heated intraoperative intraperitoneal chemotherapy with mitomycin C. AB - BACKGROUND: Treatment of carcinomatosis may involve the use of heated intraperitoneal chemotherapy; the cytotoxic solution is administered in the operating room with the abdomen open so that manual distribution results in uniform treatment. The potential risk of this procedure to the operating room personnel has not been previously investigated. METHODS: Mitomycin C was perfused through the peritoneal cavity, which was partially covered by a plastic sheet. Large volumes of air were suctioned from 5 and 35 cm above the abdominal skin edge. Urine from the surgeon and from the perfusionist were assayed. Sterile gloves worn in the operating room for manipulating the viscera during treatment were assayed for their permeability to mitomycin C. All samples were analyzed by high-performance liquid chromatography. RESULTS: Analysis of samples of operating room air and urine from 10 procedures showed no detectable levels of mitomycin C. Six tests of three different types of gloves showed a 10-fold range of mitomycin C penetration. The least permeable gloves leaked a mean of 3.8 parts per million over 90 minutes. CONCLUSIONS: No detectable safety hazard to the surgeon or other operating room personnel was demonstrated. PMID- 11888878 TI - Superficially spreading cancer of the stomach. AB - BACKGROUND: Superficially spreading cancer (SSC) of the stomach is rare and extends widely along the mucosa or submucosa of the stomach. This study was conducted to clarify the clinicopathologic characteristics and prognosis of patients with SSC. METHODS: SSC was defined as a tumor invading the mucosa or submucosa and measuring > or =5 cm in size. The clinicopathologic findings and outcomes of 36 patients with SSC were compared with those of 300 patients with early gastric cancer (EGC) measuring < or =5 cm and 271 with advanced gastric cancer measuring > or =5 cm. RESULTS: SSC was significantly different from ordinary EGC in tumor size, frequency of lymph node metastasis, lymphatic invasion, venous invasion, and stage II, III, and IV disease. The frequency of serosal invasion, lymph node metastasis, and lymphatic and venous invasions in cases of SSC was significantly lower than with advanced gastric cancer. Although tumor size of SSC evaluated before operation was smaller than that on the resected specimen, the 10-year survival rate was not different between SSC and ordinary EGC. CONCLUSIONS: SSC was characterized by high frequency of lymph node metastasis and preoperative underestimation of tumor size. SSC should be treated by a gastrectomy and lymphadenectomy with sufficient resection margin. PMID- 11888879 TI - Hurthle cell carcinoma: a 60-year experience. AB - BACKGROUND: The aim of this study was to define the clinical behavior and prognostic indicators of outcome in Hurthle cell cancer (HCC). METHODS: Diagnosis was confirmed for 56 patients with HCC treated between 1940 and 2000, who form the basis of this study. Primary end points were relapse-free survival (RFS) and disease-specific survival (DSS). Data were analyzed with the Kaplan-Meier method and by log-rank test. RESULTS: The extent of thyroid resection did not predict outcome. Recurrence was a significant predictor of tumor-related mortality. Significant adverse predictors of RFS and DSS were degree of invasion, size >4 cm, extrathyroidal extension, and initial nodal or distant metastases. The most significant predictor of outcome was extent of invasion. Eight-year RFS values for low- and high-risk groups were 100% and 24%. Corresponding rates of 8-year DSS were 100% and 58%. CONCLUSIONS: Widely invasive HCC is an aggressive malignancy that identifies patients who are at high risk for recurrence and tumor related death. Patients with HCC have a prognosis that is reliably predicted by degree of invasion, tumor size, extrathyroidal disease extension, and initial nodal or distant metastasis. Recurrence portends a poor outcome. High-risk patients and those with recurrence should be considered for adjuvant therapy. PMID- 11888880 TI - Staging laparoscopy for potentially resectable noncolorectal, nonneuroendocrine liver metastases. AB - BACKGROUND: Carefully selected patients with noncolorectal, nonneuroendocrine (NCNN) liver metastases may benefit from hepatic resection. The incidence of occult unresectable disease and the possible benefits of staging laparoscopy in these patients are not known. METHODS: From December 1997 to July 2000, staging laparoscopy was performed in 30 consecutive patients with NCNN metastases before planned open exploration and resection. Demographics, extent of preoperative imaging, operative and postoperative findings, and factors associated with laparoscopic identification of unresectable disease were analyzed. RESULTS: Twenty-four patients (80%) had a complete laparoscopic examination, and 23 had laparoscopic ultrasonography. All patients underwent preoperative computed tomography or magnetic resonance imaging, and 21 (70%) patients had 2 or more preoperative radiological studies. Overall, nine patients had unresectable disease, six of whom were identified by laparoscopy. Of the remaining 24 patients believed to have resectable disease at laparoscopy, 21 went on to a potentially curative procedure. Laparoscopy did not identify irresectability because of vascular involvement in three patients. Laparoscopy added a median of 30 minutes of operative time to those patients going on to laparotomy. CONCLUSIONS: Laparoscopy identified the majority of patients with occult unresectable disease, improved resectability, and should be routine in patients being considered for potentially curative hepatic resection. PMID- 11888881 TI - Esophageal resection for carcinoma in patients older than 70 years. AB - BACKGROUND: A larger number of older patients are presenting as candidates for esophageal resection. An aggressive surgical approach in this population is controversial. METHODS: Four hundred thirteen patients with esophageal cancer who presented to Roswell Park Cancer Institute from 1991 to 1998 were retrospectively reviewed. Clinical data, perioperative details, and postoperative courses were compared for patients older and younger than 70 years. RESULTS: One hundred forty seven patients (36%) were older than 70 years. Risk factors, clinical symptoms, histology, and stage at presentation were equivalent for both age groups. A higher percentage of patients <70 years were candidates for curative resection. There were no significant differences between groups for estimated blood loss, intraoperative transfusions, length of stay, overall morbidity, or mortality. Only postoperative myocardial infarction and atrial fibrillation were increased in the older group. Excluding stage IV disease, there was a significant and similar improvement in median survival after resection for patients both <70 years and >70 years. CONCLUSIONS: In conclusion, esophageal cancer in older patients warrants surgical resection because the benefit to the patient is the same as it is for younger patients, without a significant increase in operative morbidity or mortality. PMID- 11888882 TI - The Mouse Tumor Biology Database: a public resource for cancer genetics and pathology of the mouse. AB - Developing genetic mouse models for cancer research has been recognized as an "exceptional opportunity" by the National Cancer Institute. The establishment of bioinformatics resources to facilitate access to published and unpublished data on the genetics and pathology of cancer in different strains of the laboratory mouse is critical to developing and using mouse models of human disease. In this article, we review the Mouse Tumor Biology Database (MTB), a public resource for information on cancer genetics, epidemiology, and pathology in genetically defined mice. We outline current content, data acquisition strategies, and query mechanisms for MTB. MTB is accessible on-line at http://tumor.informatics.jax.org. PMID- 11888883 TI - Regions of H- and K-ras that provide organ specificity/potency in mammary cancer induction. AB - Organ-specific cancers with activated ras oncogenes most often are associated exclusively with only one ras isoform. For example, only H-ras activation is associated with rat mammary cancers. The mechanism underlying this specificity is mostly unknown. We have shown previously that this tissue specificity of Ras isoforms is defined by the Ras protein itself and not by differential gene expression among Ras family members. Here we show that elements in the known domains in the hypervariable region of Ras (amino acids 170-189) interact in part to control this mammary/H-Ras specificity. In addition, these in vivo mammary studies for the first time identify domains in the mostly homologous region of Ras (amino acids 1-169) that strongly influence the oncogenic potency/specificity of H-Ras. PMID- 11888884 TI - Isolation of a novel gene, CABC1, encoding a mitochondrial protein that is highly homologous to yeast activity of bc1 complex. AB - To search for p53 target genes throughout the human genome, we applied a cDNA microarray system using adenovirus-mediated transfer of p53 into p53-deficient U373MG (glioblastoma) cells. In this manner, we detected dozens of genes that appeared to be regulated by wild-type p53. We describe here characterization of one such gene, termed CABC1 [chaperone-activity of bc1 complex in Schizosaccharomyces pombe (ABC1)-like], which encodes a 647-amino acid peptide with significant sequence similarity to activity of bc1 complex (ABC1) in Arabidopsis thaliana and S. pombe. The CABC1 product was located in mitochondria, and colony-formation assays with cancer cell lines indicated its ability to suppress cell growth. Inhibition of CABC1 expression by transfection with antisense oligonucleotide significantly reduced the apoptotic response induced by wild-type p53. These results suggest that CABC1 may play an important role in mediating p53-inducible apoptosis through the mitochondrial pathway. PMID- 11888885 TI - Heregulin induces expression, ATPase activity, and nuclear localization of G3BP, a Ras signaling component, in human breast tumors. AB - We have found using differential display of mRNA that the growth factor heregulin beta 1 (HRG), a combinatorial ligand for human epidermal growth factor receptors (HERs), induced expression of G3BP, the Ras GTPase-activating protein SH3 domain binding protein, in breast cancer cells. G3BP is a downstream effector protein of Ras signaling with ATP-dependent RNase and helicase activities, which may link Ras signaling with RNA turnover and cell cycle progression. In human breast cancer cells, HRG induced G3BP mRNA and protein expression. Up-regulation of G3BP was found in MCF7 breast cancer cells overexpressing HER2. G3BP was also overexpressed in human breast tumors in parallel with HER2 overexpression and in an estrogen-independent manner, suggesting a role for G3BP in cancer progression. In addition, HRG stimulation of breast cancer cells promoted phosphorylation of G3BP and increased the association of G3BP with GTPase-activating protein, both of which are essential for G3BP activity. G3BP ATPase activity was also significantly increased by HRG treatment. Furthermore, HRG treatment resulted in G3BP translocation to the nucleus and colocalization with acetylated histone H3, a hallmark of active transcription sites. G3BP induction, phosphorylation, ATPase activity, and relocalization after HRG treatment could all be blocked by pretreatment with the anti-receptor HER2 monoclonal antibody Herceptin (trastuzumab), which may suggest additional applications for this therapeutic antibody. These findings demonstrate for the first time the receptor-dependent regulation of G3BP, a downstream effector of Ras signaling, by HRG, a growth factor with diverse functions in breast cancer cells. PMID- 11888886 TI - Clinical validation of candidate genes associated with prostate cancer progression in the CWR22 model system using tissue microarrays. AB - To explore molecular mechanisms of prostate cancer progression, we applied tissue microarrays (TMAs) to analyze expression of candidate gene targets discovered by cDNA microarray analysis of the CWR22 xenograft model system. A TMA with 544 clinical specimens from different stages of disease progression was probed by mRNA in situ hybridization and protein immunohistochemistry. There was an excellent correlation (r = 0.96; n = 16) between the expression levels of the genes in the xenografts by cDNA microarray and mRNA in situ hybridization on a TMA. One of the most highly overexpressed genes in hormone-refractory CWR22R xenografts was the S100P gene. This gene, coding for a calcium signaling molecule implicated in the loss of senescence, was also significantly associated with progression in clinical tumors by TMA analysis (P < 0.001), suggesting dysregulation of this pathway in hormone-refractory and metastatic prostate cancers. Conversely, two genes that were down-regulated during tumor progression in the CWR22 model system were validated in vivo: crystallin mu (CRYM) and a LIM domain protein LMO4 both showed significantly lower mRNA levels in hormone refractory tumors as compared with primary tumors (P < 0.001). These results illustrate a strategy for rapid clinical validation at the mRNA and protein level of gene targets found to be differentially expressed in cDNA microarray experiments of model systems of cancer. PMID- 11888887 TI - Cellular localization and function of Fas ligand (CD95L) in tumors. AB - The localization of CD95L in different cell types within tumors has not been well defined, and its role in tumor growth is uncertain. In this study, CD95L expression and its contribution to tumor growth are evaluated using genetic polymorphisms and adoptive transfer in genetically deficient mice. CD95L was detected in tumors in vivo at levels up to 10(4)-fold higher than those in cell culture and predominantly in host tumor-infiltrating macrophages, but not in tumor cells. Adoptive transfer into genetically deficient mice revealed that host CD95-CD95L function did not alter tumor growth, demonstrating that CD95L alone does not affect tumor growth. PMID- 11888888 TI - Treatment of ovarian cancer with a tropism modified oncolytic adenovirus. AB - Ad5-Delta 24RGD is an adenovirus that is selectively replication competent in cells defective in the Rb/p16 pathway, such as ovarian cancer cells. The fiber of Ad5-Delta 24RGD contains an integrin binding RGD-4C motif, allowing Coxsackie adenovirus receptor-independent infection of cancer cells. Oncolysis of cell lines was similar to that of a wild-type control, and replication in primary tumor material was shown using a novel three-dimensional spheroid model. Finally, an orthotopic murine model of peritoneally disseminated ovarian cancer was used to test i.p. administration to tumor-bearing animals. Injection of the agent resulted in eradication of i.p. disease, whereas control animals expired (P < 0.0001). These results suggest that Ad5-Delta 24RGD could be useful for treatment of ovarian cancer in humans. PMID- 11888889 TI - The flt-1 promoter for transcriptional targeting of teratocarcinoma. AB - Flt-1, a receptor for vascular endothelial growth factor, is known to display dysregulated expression in both tumor vasculature and tumor cells per se, suggesting that the flt-1 promoter might be a useful candidate to achieve tumor specific transgene expression. In addition, adenoviral vectors containing transgenes under the control of the flt-1 promoter achieve very low levels of expression in the normal liver, the major organ responsible for blood clearance of adenoviruses and inadvertent transgene-related toxicity. Thus, we assessed the ability of adenoviral vectors containing the flt-1 promoter to achieve transgene expression in a range of gynecological and breast tumor lines. High transgene expression levels were detected in teratocarcinoma lines, correlating with levels of flt-1 mRNA. These results suggest that the flt-1 promoter could be useful for transcriptionally targeted gene expression to teratocarcinoma, and that evaluation in other flt-1-positive tumors is warranted. PMID- 11888890 TI - Mouse ING1 homologue, a protein interacting with A1, enhances cell death and is inhibited by A1 in mammary epithelial cells. AB - We cloned mouse ING1 homologue (mINGh), an A1/Bfl-1-interacting protein, from mouse mammary glands using a yeast two-hybrid assay and unexpectedly found four splicing variants of mINGh by reverse transcription-PCR assay and sequence analysis. The alternative splicing variants were mINGh-S, mINGh-M, mINGh-L, and mINGh-L2 encoding 171, 248, 166, and 227 amino acids, respectively. Cell death of HC11 cells, induced by serum starvation, was enhanced by mINGhs, and the action of mINGhs was inhibited by A1 protein. These results indicate that A1 can inhibit cell death not only via the well known pathway related to the Bcl-2 family but also through direct interaction with mINGh in mammary epithelial cells. PMID- 11888891 TI - Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-x(L). AB - Homology-directed repair (HDR) of DNA double-strand breaks (DSBs) contributes to the maintenance of genomic stability in rodent cells, and it has been assumed that HDR is of similar importance in DSB repair in human cells. However, some outcomes of homologous recombination can be deleterious, suggesting that factors exist to regulate HDR. We demonstrated previously that overexpression of BCL-2 or BCL-x(L) enhanced the frequency of X-ray-induced TK1 mutations, including loss of heterozygosity events presumed to arise by mitotic recombination. The present study was designed to test whether HDR is a prominent DSB repair pathway in human cells and to determine whether ectopic expression of BCL-x(L) affects HDR. Using TK6-neo cells, we find that a single DSB in an integrated HDR reporter stimulates gene conversion 40-50-fold, demonstrating efficient DSB repair by gene conversion in human cells. Significantly, DSB-induced gene conversion events are 3-4-fold more frequent in TK6 cells that stably overexpress the antiapoptotic protein BCL X(L). Thus, HDR plays an important role in maintaining genomic integrity in human cells, and ectopic expression of BCL-x(L) enhances HDR of DSBs. This is the first study to highlight a function for BCL-x(L) in modulating DSB repair in human cells. PMID- 11888892 TI - Mutation profiling of mismatch repair-deficient colorectal cncers using an in silico genome scan to identify coding microsatellites. AB - Human colorectal, endometrial, and gastric cancers with defective DNA mismatch repair (MMR) have microsatellite instability, a unique molecular alteration characterized by widespread frameshift mutations of repetitive DNA sequences. We developed "Kangaroo," a bioinformatics program for searches in nucleotide and protein sequence databases, and performed an in silico genome scan for DNA coding microsatellites that may have novel mutations in MMR-deficient cancers. Examination of 29 previously untested coding polyadenines revealed widespread mutations in MMR-deficient colorectal cancers, with the highest frequencies in ERCC5, CASP8AP2, p72, RAD50, CDC25, RECQL1, CBF2, RACK7, GRK4, and DNAPK (range, 10-33%). This algorithm allows comprehensive mutation profiling of MMR-deficient cancers, an important step in understanding the pathogenesis of these neoplasms. PMID- 11888893 TI - Stanniocalcin 2 is an estrogen-responsive gene coexpressed with the estrogen receptor in human breast cancer. AB - Differences in gene expression are likely to explain the phenotypic variation between hormone-responsive and hormone-unresponsive breast cancers. In this study, DNA microarray analysis of approximately 10,000 known genes and 25,000 expressed sequence tag clusters was performed to identify genes induced by estrogen and repressed by the pure antiestrogen ICI 182 780 in vitro that correlated with estrogen receptor (ER) expression in primary breast carcinomas in vivo. Stanniocalcin (STC) 2 was identified as one of the genes that fulfilled these criteria. DNA microarray hybridization showed a 3-fold induction of STC2 mRNA expression in MCF-7 cells in < or = 3 h of estrogen exposure and a 3-fold repression in the presence of antiestrogen (one-way ANOVA, P < 0.0005). In 13 ER positive and 12 ER-negative breast carcinomas, the microarray-derived mRNA levels observed for STC2 correlated with tumor ER mRNA (Pearson's correlation, r = 0.85; P < 0.0001) and ER protein status (Spearman's rank correlation, r = 0.73; P < 0.0001). The expression profile of STC2 was further confirmed by in situ hybridization and immunohistochemistry on a larger cohort of 236 unselected breast carcinomas using tissue microarrays. STC2 mRNA and protein expression were found to be associated with tumor ER status (Fisher's exact test, P < 0.005). The related gene, STC1, was also examined and shown to be associated with ER status in breast carcinomas (Fisher's exact test, P < 0.05). This study demonstrates the feasibility of using global gene expression data derived from an in vitro model to pinpoint novel estrogen-responsive genes of potential clinical relevance. PMID- 11888894 TI - Complete genetic suppression of polyp formation and reduction of CpG-island hypermethylation in Apc(Min/+) Dnmt1-hypomorphic Mice. AB - Promoter CpG island hypermethylation of critical genes is thought to play an important role in human colorectal tumorigenesis. In this study, we show that low levels of CpG island methylation occur in the normal intestinal mucosa of Apc(Min/+) mice and are increased in Multiple Intestinal Metaplasia (Min) polyps. We examined the interaction between CpG island hypermethylation and tumorigenesis by genetically modulating expression levels of the predominant DNA methyltransferase, Dnmt1, in Apc(Min/+) mice. We show that a combination of Dnmt1 hypomorphic alleles results in the complete suppression of polyp formation and an accompanying reduction in the frequency of CpG island methylation in both the normal intestinal mucosa and intestinal adenomas. These results suggest that sufficient DNA methyltransferase expression is a prerequisite for polyp formation and that hypomorphic alleles of Dnmt1 are not merely genetic modifiers but the first identified true genetic suppressors of the Min phenotype. PMID- 11888895 TI - Differential gene expression profiles of Jnk1- and Jnk2-deficient murine fibroblast cells. AB - c-Jun NH(2)-terminal kinase (JNK) 1 and JNK2 have been assumed to complement each other and mediate the same or similar biological functions. However, our recent reports indicated that 7,12-dimethylbenz(a)anthracene/12-O-tetradecanoylphorbol 13-acetate-induced tumor development is suppressed in Jnk2 knockout mice but enhanced in Jnk1 knockout mice. In the present work, primary embryo cells were isolated from wild-type, Jnk1(-/-) and Jnk2(-/-) mice and used for cDNA microarray analysis. The patterns of gene expression in Jnk1(-/-), Jnk2(-/-), and wild-type cells are different. After 12-O-tetradecanoylphorbol-13-acetate treatment, the changes in the gene expression profiles in three different kinds of cells appear to agree with the differences in susceptibility to tumorigenesis of each respective animal model. These results suggest that JNK1 and JNK2 proteins have different roles in modulating cell function. PMID- 11888896 TI - Expression of p14ARF overcomes tumor resistance to p53. AB - Tumors without p53 mutation are often resistant to p53 gene therapy. We examined the mechanism using p53-resistant A549 cells and p53-sensitive H1299 cells. We found that p53 delivered by adenovirus is poorly expressed in A549 (ARF-null) cells but efficiently expressed in H1299 cells (ARF-positive). Strong p53 expression and apoptosis can be achieved in A549 cells using a p53 mutant resistant to degradation by MDM2 or by coexpression of ARF. The results suggest that enhanced MDM2 activity attributable to loss of ARF contributes to p53 resistance. Surprisingly, tumor cell lines with MDM2 gene amplification are still deficient for ARF expression, suggesting that MDM2 amplification does not substitute for ARF inactivation during tumor development. PMID- 11888897 TI - Inhibition of cancer cell growth by BRCA2. AB - The breast cancer susceptibility gene BRCA2 has been suggested to function as a "caretaker" of the genome. Cells without wild-type BRCA2 are deficient in repairing DNA damage. However, whether BRCA2 can also suppress oncogenesis by regulating cell proliferation remains to be determined. To address this question, the expression of wild-type BRCA2 protein was reconstituted, in an either constitutive or regulated manner, in the pancreatic cancer cell line Capan-1, which expresses only a mutant BRCA2. Expression of wild-type BRCA2 inhibited cell proliferation in culture and suppressed tumor growth in animals. Our results showed that, in addition to the DNA repair function, BRCA2 also suppresses tumor development by inhibiting cancer cell growth. PMID- 11888898 TI - Intratumoral lymphangiogenesis and lymph node metastasis in head and neck cancer. AB - How tumors access and spread via the lymphatics is not understood. Although it is clear that dissemination via the blood system involves hemangiogenesis, it is uncertain whether tumors also induce lymphangiogenesis or simply invade existing peritumoral vessels. To address the issue we quantitated tumor lymph vessels in archival specimens of head and neck cancer by immunostaining for the recently described lymphatic endothelial marker LYVE-1, the vascular endothelial marker CD34, and the pKi67 proliferation marker, correlating lymph vessel density and proliferation index with clinical and pathological variables. Discrete "hotspots" of intratumoral small proliferating lymphatics were observed in all carcinomas, and a high intratumoral lymph vessel density was associated with neck node metastases (n = 23; P = 0.027) and an infiltrating margin of tumor invasion (P = 0.046) in the oropharyngeal subgroup. Quantitation of the lymphangiogenic growth factor vascular endothelial growth factor C by real-time PCR and immunohistochemistry revealed higher levels of mRNA in tumor tissue than in normal samples (n = 8; P = 0.017), but no obvious correlation with intratumoral lymphatics. Our results provide new evidence that proliferating lymphatics can occur in human cancers and may in some cases contribute to lymph node metastasis. PMID- 11888899 TI - Identification of a novel function for 67-kDa laminin receptor: increase in laminin degradation rate and release of motility fragments. AB - The 67-kDa laminin receptor (67LR) is a high-affinity laminin-binding protein that is overexpressed on the tumor cell surface in a variety of cancers. We report here that the 67LR molecule also functions in the proteolytic cleavage of laminin-1, a relevant event in basement membrane degradation and tumor dissemination. In the presence of a synthetic peptide (peptide G) corresponding to the 67LR laminin binding site, the rate of laminin-1 degradation by the cysteine proteinase cathepsin B was significantly increased, and a new proteolytic fragment particularly active in in vitro cell migration assays was generated. The YIGSR peptide, corresponding to the 67LR binding site on laminin 1, blocked the peptide G-dependent proteolytic degradation. Our results shed light on the mechanism by which an adhesion receptor such as the 67LR plays a major role in tumor aggressiveness and metastasis. PMID- 11888900 TI - Relation of hypoxia-inducible factor-2 alpha (HIF-2 alpha) expression in tumor infiltrative macrophages to tumor angiogenesis and the oxidative thymidine phosphorylase pathway in Human breast cancer. AB - Tumor-associated macrophages (TAMs) produce angiogenic factors and in breast cancer are associated with high vascular grade and poor survival. TAMs preferentially migrate to hypoxic areas within tumors and strongly express hypoxia-inducible factor (HIF)-2 alpha. This study examined whether HIF-2 alpha was involved in TAM angiogenic activation by correlating its expression with tumor microvessel density as a marker of angiogenesis, and other tumor variables, in a series of human primary invasive breast carcinomas. A correlation was found between high TAM HIF-2 alpha and high tumor vascularity (P < 0.0001), as well as high tumor grade (P = 0.007). The relation of HIF-2 alpha expression to a recently described oxygen-dependent pathway of angiogenesis was also studied, and an inverse relationship was found between TAM HIF-2 alpha and tumor thymidine phosphorylase expression (P = 0.02). These results suggest that TAM HIF-2 signaling may be a useful target for future antiangiogenic strategies but show that tumors use both oxygen-dependent and oxygen deficiency-regulated pathways for angiogenesis. Thus, combined blockade of pathways and careful assessment of these pathways in trials are necessary. PMID- 11888901 TI - Albumin adducts of benzene oxide and 1,4-benzoquinone as measures of human benzene metabolism. AB - Albumin adducts of benzene oxide (BO-Alb) and 1,4-benzoquinone (1,4-BQ-Alb) were investigated among 134 workers exposed to benzene and 51 unexposed controls in Tianjin, China. Concentrations of both adducts increased with benzene exposure [range = 0.07-46.6 parts/million (ppm); median = 3.55 ppm] and with urinary cotinine. Adduct levels were less than proportional to benzene exposure, suggesting saturable CYP 2E1 metabolism of benzene. Because the transition from linear to saturable metabolism began at approximately 1 ppm, the common assumption of linear kinetics at much higher benzene exposures could lead to substantial underestimation of leukemia risks. Adduct levels were generally lower in older workers, indicating that CYP 2E1 metabolism diminished with age, at approximately 2%/year of life. The ratio of 1,4-BQ-Alb:BO-Alb decreased with age and coexposure to toluene, and increased with alcohol consumption. This indicates that factors affecting CYP 2E1 metabolism exerted a greater role on production of 1,4-BQ than BO, presumably because of the second oxidation step from phenol to hydroquinone. The adduct ratio was also positively associated with urinary cotinine, suggesting that both benzene and hydroquinone from cigarette smoke affected adduct levels. Results of a limited time course study of 11 subjects indicated moderate chemical instability of 1,4-BQ-Alb (half life = 13.5 days compared with 21 days for normal Alb turnover), whereas no evidence of instability of BO-Alb was observed. This study illustrates that Alb adducts can be used to investigate the dispositions of reactive metabolites of procarcinogens in humans, provided that exposures are adequately characterized in the month preceding blood collection. PMID- 11888902 TI - Suppression of UV carcinogenesis by difluoromethylornithine in nucleotide excision repair-deficient Xpa knockout mice. AB - Xeroderma pigmentosum (XP) patients are deficient in nucleotide excision repair (NER) because of mutations in one of the genes coding for NER enzymes. This results predominantly in high frequency of UV-induced skin tumors at an early age; the most severe phenotype is found in patients of complementation group A (XPA). However, in a subset of these XPA patients no skin tumors appear, even at advanced age. Fibroblasts of this subset of patients are not capable of raising UV-induced enhanced reactivation (ER) of viruses and up-regulation of ornithine decarboxylase (ODC). We hypothesized that prevention of ODC induction would protect NER-deficient patients from cancer. To simulate the situation in XPA patients, we used a hairless Xpa knockout mouse model and down-regulated the ODC activity by difluoromethylornithine (DFMO) administered in the drinking water. The DFMO treatment significantly suppressed UV-induced carcinogenesis. In a crossover study, we additionally found that discontinuation of the DFMO treatment resulted in a rapid appearance of skin tumors, up to levels found in mice not treated with DFMO. Late-stage DFMO treatment significantly reduced the number of carcinomas by a factor of 2-3, and it appeared to select for carcinomas with high ODC activity. These results indicate that DFMO suppresses the outgrowth but not the initiation of UV-induced tumors. The DFMO treatment reduced the tumor load but did not offer the Xpa knockout mice full protection against UV carcinogenesis. PMID- 11888903 TI - Deficiency of c-Jun-NH(2)-terminal kinase-1 in mice enhances skin tumor development by 12-O-tetradecanoylphorbol-13-acetate. AB - The c-Jun NH(2)-terminal kinase (JNK) has been implicated in regulating cell survival, apoptosis, and transformation. However, the distinct role of JNK isoforms in regulating tumor development is not yet clear. We have found previously that skin tumor formation induced by the tumor promoter, 12-O tetradecanoylphorbol-13-acetate (TPA), is suppressed in JNK2-deficient (Jnk2(-/ )) mice. Here, we show that JNK1-deficient (Jnk1(-/-)) mice are more susceptible to TPA-induced skin tumor development than wild-type mice. The rate of tumor development in Jnk1(-/-) mice was significantly more rapid than that observed in wild-type mice (P < 0.0001). At the end of 33 weeks of TPA promotion, the number of skin tumors and tumors >1.5 mm in diameter per mouse in Jnk1(-/-) mice was significantly increased by 71% (P < 0.03) and 82% (P < 0.03), respectively, relative to the wild-type mice. Furthermore, the carcinoma incidence and the number of carcinomas per mouse were also higher in Jnk1(-/-) mice. Strikingly, Jnk1(-/-) mouse skin was more sensitive to TPA-induced AP-1 DNA binding activity and phosphorylation of extracellular signal-regulated kinases and Akt, which are two important survival signaling components. These results suggest that JNK1 is a crucial suppressor of skin tumor development. PMID- 11888904 TI - Defective repair of 8-hydroxyguanine in mitochondria of MCF-7 and MDA-MB-468 human breast cancer cell lines. AB - Breast cancer is one of the major causes of mortality among women in the United States. Although the causes of breast cancer remain unclear, it has been speculated that DNA base damage may lead to mutations that subsequently can be carcinogenic. Recently, defective oxidative DNA damage repair has been implicated in breast tumorigenesis. The major oxidative DNA lesion, 8-hydroxyguanine (8 oxoG), is increased in breast cancer, suggesting that this lesion may play a crucial role in the etiology of breast cancer. However, it is not known whether the repair of 8-oxoG or other oxidative base lesions is altered during breast carcinogenesis. We examined the ability of nuclear and mitochondrial extracts of two human breast cancer cell lines, MCF-7 and MDA-MB-468, to repair 8-oxoG lesion. We report that mitochondrial extracts from the two breast cancer cell lines are defective in the base excision repair of 8-oxoG relative to two noncancer cell lines. We also show that the incision activity of 8-oxoG was significantly lower in mitochondrial than in nuclear extracts in the breast cancer cell lines. The defective mitochondrial repair activity was not attributable to lower levels of human 8-hydroxyguanine DNA glycosylase, the base excision repair enzyme known to incise 8-oxoG in DNA. The repair of thymine glycol, another major oxidative DNA base lesion that blocks transcription and causes cell death, was similar in cancer and noncancer cells. Furthermore, nuclear extracts incised thymine glycol with a much higher efficiency than 8 oxoG. These data provide evidence for defective repair of 8-oxoG in mitochondria of MCF-7 and MDA-MB-468 breast cancer cell lines. These results may implicate 8 oxoG repair mechanisms in mitochondria of certain breast cancers. PMID- 11888905 TI - Magnetic field exposure increases cell proliferation but does not affect melatonin levels in the mammary gland of female Sprague Dawley rats. AB - In line with the possible relationship between electric power and breast cancer risk as well as the underlying "melatonin hypothesis," we have shown previously (Thun-Battersby et al., Cancer Res., 59: 3627-3633, 1999) that 50-Hz magnetic fields (MFs) of low (100 microTesla) flux density enhance mammary gland tumor development and growth in the 7,12-dimethylbenz(a)anthracene model of breast cancer in female Sprague Dawley rats. On the basis of the melatonin hypothesis and previous observations of induction of ornithine decarboxylase in response to MF, we proposed that the effect of MF exposure on mammary carcinogenesis is related to enhanced proliferation of the mammary epithelium. The objective of the present study was to directly assess this proposal by the use of proliferation markers. Female Sprague Dawley rats were MF or sham exposed for 2 weeks at a flux density of 100 microTesla. Proliferation of epithelial cells in the mammary tissue and adjacent skin was examined by in vivo labeling of proliferating cells with bromodeoxyuridine (BrdUrd) and in situ labeling of the nuclear proliferation associated Ki-67 protein by the antibody MIB-5. Furthermore, melatonin levels were determined after MF or sham exposure in the pineal gland and directly in the mammary tissue. In additional experiments, the tumor promoter 12-O tetradecanoylphorbol-13-acetate was used for comparison with the effects of MF exposure. MF exposure significantly enhanced BrdUrd and Ki-67 labeling in the mammary epithelium, indicating a marked increase in cell proliferation. The most pronounced effect on proliferation was seen in the cranial thoracic (or cervical) mammary complexes, in which we previously had seen the most marked effects of MF exposure on mammary carcinogenesis. In contrast to the melatonin hypothesis, melatonin levels in pineal or mammary glands were not affected by MF exposure. Topical application of 12-O-tetradecanoylphorbol-13-acetate increased BrdUrd and Ki-67 labeling in epithelial cells of the skin, particularly in hair follicles, but not in the mammary tissue. The data demonstrate that MF exposure results in an increased proliferative activity of the mammary epithelium, which is a likely explanation for the cocarcinogenic or tumor promoting effects of MF exposure observed previously by us in the 7,12-dimethylbenz(a)anthracene model of breast cancer. PMID- 11888906 TI - Induction of IgG subclass responses in colorectal carcinoma patients vaccinated with recombinant carcinoembryonic antigen. AB - There is scanty information on the IgG subclass response after vaccination against cancer antigens. The induction and development of the IgG subclass responses in 18 colorectal carcinoma patients vaccinated s.c. seven times with recombinant human carcinoembryonic antigen (rhCEA) over a 12-month period were analyzed by ELISA. The patients were followed for 3 years. Four rhCEA doses were used, and half of the patients also received granulocyte macrophage-colony stimulating factor (GM-CSF) as an adjuvant. Anti-rhCEA-specific IgG1 and IgG4 responses and, to a lesser degree, IgG2 responses were markedly enhanced by concomitant GM-CSF administration, whereas the antigen dose was of minor importance. Almost no IgG3 response was observed. A significant antibody response was noted within the first weeks for IgG1 and IgG2 but noted several months later for IgG4. The responses gradually increased by repeated immunizations and peaked around 12 months for IgG1 and a few months later for IgG2 and IgG4. A sustained but reduced response was noted for these three subclasses at 24 and 36 months. Interestingly, there was a gradual shift from a predominant IgG1 response at 6 months to an IgG4 response at 15 months. No significant change in total concentrations of the four IgG subclasses was observed comparing prevaccination concentrations with concentrations at 12 months, indicating an antigen-specific effect of GM-CSF administration on the anti-rhCEA response. The clinical significance of the individual IgG subclass antibodies for tumor response is not clear and requires additional studies. PMID- 11888907 TI - Toremifene prevents prostate cancer in the transgenic adenocarcinoma of mouse prostate model. AB - The chemopreventive efficacy of toremifene, an antiestrogen, was evaluated in the transgenic adenocarcinoma of mouse prostate (TRAMP) model. TRAMP mice were segregated into three groups: (a) the low-dose toremifene group (6.6 mg/kg/day); (b) the high-dose toremifene group (33 mg/kg/day); and (c) the control placebo group. Efficacy of treatment was measured by the absence of palpable tumor. To extend these studies using more sensitive techniques, TRAMP mice were then treated with placebo, flutamide (an antiandrogen; 33 mg/kg/day), or toremifene (10 mg/kg/day). Animals from each treatment group were sacrificed at 7, 10, 15, 20, 25, and 30 weeks of age, and prostate tissues and seminal vesicles were harvested. Tissues from animals (n = 5) in each group were evaluated by wholemount dissections of genitourinary tracts, histology, immunohistochemistry, and Western blot analyses. Blood was pooled per group to measure estradiol and testosterone hormonal levels. Tumors formed at week 17 in the placebo group (n = 10), at week 21 in the high-dose toremifene group (n = 12), and at week 29 in the low-dose toremifene group (n = 12). This represents an increased tumor latency of up to 12 weeks. By 33 weeks, all animals in the placebo group had tumors compared with only 35% of the animals treated with toremifene. Although both flutamide and toremifene decreased tumor incidence compared with the placebo, toremifene was more effective than flutamide. High-grade prostatic intraepithelial neoplasia was observed in animals in the placebo group, but not in animals treated with toremifene. Moreover, toremifene-treated animals had prolonged survival compared with placebo-treated animals. By 33 weeks of age, 100% of the placebo-treated animals had developed palpable tumors and died, whereas 60% of the toremifene treated animals were tumor free. T antigen levels in the prostate of toremifene treated animals were similar to those of placebo-treated, age-matched animals. Whereas serum estradiol levels remained unchanged, the total and free testosterone levels were elevated in the toremifene-treated group. Toremifene treatment did not affect androgen receptor levels. Because toremifene prevented prostate cancer in a milieu of elevated blood free testosterone levels with no change in prostate androgen receptor expression, the mechanism of toremifene's chemopreventive activity may be through nonandrogenic pathways, such as estrogen receptor signaling. PMID- 11888908 TI - Gene-environment interaction for the ERCC2 polymorphisms and cumulative cigarette smoking exposure in lung cancer. AB - Excision repair cross-complementing group 2 (ERCC2), a major DNA repair protein, is involved in nucleotide excision repair and basal transcription. The ERCC2 polymorphisms have been associated with altered DNA repair capacity. We investigated two ERCC2 polymorphisms, Asp312Asn and Lys751Gln, in 1092 Caucasian lung cancer patients and 1240 spouse and friend controls. The results were analyzed using generalized additive models and logistic regression, adjusting for relevant covariates. The overall adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were 1.47 (95% CI, 1.1-2.0) for the Asp312Asn polymorphism (Asn/Asn versus Asp/Asp) and 1.06 (95% CI, 0.8-1.4) for the Lys751Gln polymorphism (Gln/Gln versus Lys/Lys). Gene-smoking interaction analyses revealed that the adjusted ORs for each of the two polymorphisms decreased significantly as pack-years increased. When comparing individuals with Asn/Asn + Gln/Gln versus individuals with Asp/Asp + Lys/Lys, the fitted ORs (95% CIs) were 2.56 (95% CI, 1.3-5.0) in nonsmokers and 0.69 (95% CI, 0.4-1.2) in heavy smokers (80 pack years; P < 0.01 for the interaction term). Consistent and robust results were found when models incorporated different definitions of cumulative cigarette smoking. A stronger gene-smoking interaction was observed for the Asp312Asn polymorphism than for the Lys751Gln polymorphism. In conclusion, cumulative cigarette smoking modifies the associations between ERCC2 polymorphisms and lung cancer risk. PMID- 11888909 TI - Cells designed to deliver anticancer drugs by apoptosis. AB - We describe a new drug delivery strategy that uses genetically engineered endothelial cells (ECs) to deliver drugs to tumor cells by apoptosis. Immortalized ECs were genetically engineered to express a flk-1:fas fusion protein. When exposed to the flk-1 ligand, vascular endothelial growth factor (VEGF), which is overexpressed by many tumors, these cells underwent extensive apoptosis. These apoptotic ECs, when loaded with drug, promote drug delivery by phagocytosis of drug-loaded apoptotic bodies by the tumor cells and by increased drug transport through the more permeable apoptotic membrane. In the current study, severe combined immunodeficient mice bearing s.c. tumors that expressed high levels of VEGF were treated either intratumorally or i.v. every 4 days for a total of five doses with saline control, free Taxol, and immortalized ECs (imECs) expressing the flk-1:fas fusion protein (imEC/HFF) loaded with Taxol (imEC/HFF T). Intratumoral treatments also included imEC/HFF and imECs loaded with Taxol (imEC-T). Tumor size was monitored for a minimum of 44 days. Whether administered intratumorally or i.v., imEC/HFF-T cells produced greater inhibition of tumor growth than all other treatments, including Taxol. It was noteworthy that 5 of 16 of the imEC/HFF-T-treated animals were tumor free at the termination of the studies, compared with 2 of 16 animals treated with Taxol. A cell distribution experiment showed that flk-1:fas fusion protein expression ECs as well as parental ECs accumulated in tumor and spleen with the highest level, followed by liver, lung, kidney, and brain. Significant apoptosis of flk-1:fas expression cells was observed in tumor, apparently driven by VEGF secreted from tumor cells. Apoptosis-induced drug delivery offers a new avenue for targeted drug delivery research that uses biological control mechanisms. PMID- 11888910 TI - Peripheral benzodiazepine receptor ligands reverse apoptosis resistance of cancer cells in vitro and in vivo. AB - The mitochondrial peripheral benzodiazepine receptor (mPBR) is involved in a functional structure designated as the permeability transition pore, which controls apoptosis. Binding of Fas/APO-1/CD95 triggers a prototypic apoptosis inducing pathway. Using four different human tumor cell lines (T-cell Jurkat, neuroblastoma SHEP, osteosarcoma 143N2, and glioblastoma SNB79 cell lines), all of which express CD95 and mPBR, we investigated the potential role of mPBR ligands in CD95-induced apoptosis. We show that, in vitro, the three mPBR ligands tested (RO5-4864, PK11195, and diazepam) enhanced apoptosis induced by anti-CD95 antibody in Jurkat cells, as demonstrated by mitochondrial transmembrane potential drop and DNA fragmentation. In contrast, RO5-4864, but not PK11195 or diazepam, enhanced anti-CD95 apoptosis in all other cell lines. These effects were obtained in Bcl-2-overexpressing SHEP cell lines, but not in Bcl-X(L) SHEP cell lines. Enhancement of anti-CD95 antibody-induced apoptosis by RO5-4864 was characterized by an increased mitochondrial release of cytochrome c and Smac/DIABLO proteins and an enhanced activation of caspases 9 and 3, suggesting a mitochondrion-dependent mechanism. Preincubation of cells with the different mPBR ligands or anti-CD95 did not affect the levels of expression of either mPBR or CD95. In vivo, we found that the RO5-4864 mPBR ligand significantly increased the growth inhibition induced by two chemotherapeutic agents, etoposide and ifosfamide, using two human small cell lung cancers xenografted into nude mice. Peripheral benzodiazepine receptor ligands may therefore act as chemosensitizing agents for the treatment of human neoplasms. PMID- 11888911 TI - Nuclear magnetic resonance-visible lipids induced by cationic lipophilic chemotherapeutic agents are accompanied by increased lipid droplet formation and damaged mitochondria. AB - Proton nuclear magnetic resonance (NMR) spectroscopy, histological lipid staining, and electron microscopy were used to assess the biochemical and structural changes induced by treating the cultured human breast cell line HBL 100 with the cationic lipophilic phosphonium salts p-(triphenylphosphoniummethyl) benzaldehyde chloride (drug A) and [4-(hydrazinocarboxy)-1-butyl] tris-(4 dimethylaminophenyl) phosphonium chloride (drug B). The major biochemical change detected by (1)H NMR in drug-treated cells was a significant time- and concentration-dependent increase in lipid acyl chain resonances arising from mobile lipids. The amount of NMR-visible lipid strongly correlated with morphometric measurements of oil red O-staining lipid detected in the cytoplasm by light microscopy. Ultrastructural investigations revealed substantial damage to mitochondria and the progressive development of lipid droplets accompanied by end-stage autophagic vacuoles, in the form of densely staining myelinoid bodies, after treatment of HBL-100 cells with drug B at the IC(50). No apparent increase in acid phosphatase activity was observed using electron microscopy, indicating that the accumulation of phospholipids in myelinoid bodies may result from substrate inundation of the lysosome, rather than increased lysosomal activity. These results indicate a potential role for lysosomal lipid catabolism in the formation of NMR-visible lipids in models of cytotoxic insult. PMID- 11888912 TI - Perifosine, a novel alkylphospholipid, induces p21(WAF1) expression in squamous carcinoma cells through a p53-independent pathway, leading to loss in cyclin dependent kinase activity and cell cycle arrest. AB - Alkylphospholipids (ALKs) are a novel class of antineoplastic compounds that display potent antiproliferative activity against several in vitro and in vivo human tumor models. However, the mechanism by which these agents exert this desired effect is still unclear. In this study, we investigated the effect of perifosine, a p.o.-bioavailable ALK, on the cell cycle kinetics of immortalized keratinocytes (HaCaT) as well as head and neck squamous carcinoma cells. All cells were sensitive to the antiproliferative properties of perifosine with an IC(50) of similar0.6-8.9 microM. Cell cycle arrest at the G(1)-S and G(2)-M boundaries was observed in HN12, HN30, and HaCaT cells independent of p53 function, and this effect was preceded by loss in cdc2 and cyclin-dependent kinase (cdk) 2 activity. Analysis of cdk complexes in vitro demonstrated that perifosine, up to 20 microM, did not directly interfere with these enzymes. However, aphidicolin-synchronized HN12 cells released in the presence of perifosine (10 microM) demonstrated increased expression of total p21(WAF1) and increased association of p21(WAF1) with cyclin-cdk complexes resulting in reduced cdc2 activity. HCT116 isogenic cell lines were used to assess the role of p21(WAF1) induction by perifosine. This compound (20 microM) induced both G(1)-S and G(2)-M cell cycle arrest, together with p21(WAF1) expression in both p53 wild type and p53(-/-) clones. By contrast, p21(-/-) variants demonstrated no p21(WAF1) induction or cell cycle arrest. Similar results were obtained with other ALK congeners (miltefosine and edelfosine). These data, therefore, indicate that perifosine blocks cell cycle progression of head and neck squamous carcinoma cells at G(1)-S and G(2)-M by inducing p21(WAF1), irrespective of p53 function, and may be exploited clinically because the majority of human malignancies harbor p53 mutations. PMID- 11888913 TI - Marked prevention of tumor growth and metastasis by a novel immunosuppressive agent, FTY720, in mouse breast cancer models. AB - FTY720 is a unique immunosuppressive agent that exerts its activity by inducing apoptosis in lymphocytes. We conducted the present study to investigate the effects of FTY720 on cancer growth and metastasis, as well as its mechanism of action. In vitro treatment with FTY720 induced dramatic cancer cell apoptosis in a mouse breast cancer cell line, JygMC(A). Electron microscopy revealed distinct changes on the cell surface with decreased filopodias and microvilli in cancer cells treated with FTY720 at 2 microM and clear evidence of apoptosis at 10 microM. Interestingly, the effect of FTY720 was significantly less in the normal fibroblasts than in the cancer cells, indicating greater susceptibility of cancer cells to the agent. We then tested the in vivo effect of FTY720 in a mouse breast cancer model created by inoculating JygMC(A) cells (s.c.) in the flank region of BALB/c-nu/nu mice at three different dosages (2, 5, and 10 mg/kg/day; n = 30/group). Tumor growth was markedly suppressed at a dosage of 5 mg/kg or more without notable side effects. In addition, tumor metastasis, which was dramatically evident in control mice, was significantly prevented even at a low dose (2 mg/kg/day), resulting in a significant prolongation of animal survival. These data led us to additionally investigate the mechanism of action, especially the prevention of metastasis at a low dose. FTY720 treatment at 2 microM caused a remarkable cytoskeletal change with deformed and decreased filopodias in cancer cells. In addition, it significantly decreased the ability of cancer cells to adhere and migrate to extracellular matrix components, and markedly reduced the expression of integrins on the cancer cell surface. These results indicate that FTY720 is a potent anticancer agent that induces cancer cell apoptosis and is markedly effective for prevention of metastasis. The changes of cellular structure with reduction of integrin expression may be one of its underlying mechanisms of action. PMID- 11888914 TI - Subcellular localization of NAD(P)H:quinone oxidoreductase 1 in human cancer cells. AB - NAD(P)H:quinone oxidoreductase 1 (NQO1) is implicated in both chemoprevention and bioactivation of DNA-damaging antitumor agents. NQO1 is mainly cytosolic, but distribution in other cellular compartments, particularly in tumor cells, is poorly defined. Nuclear NQO1 in HT29 human colon carcinoma and H661 human non small cell lung cancer cells was observed using both confocal microscopy and immunoelectron microscopy. NQO1 was not detected in mitochondria, golgi, or endoplasmic reticulum. In addition, purified intact nuclei from HT29 cells contained immunoreactive NQO1, which was catalytically active as determined by conventional activity assay. In summary, we have confirmed the presence of nuclear NQO1, which has implications for chemoprotection and bioactivation of DNA damaging antitumor agents. PMID- 11888915 TI - Quantitation of bystander effects in nitroreductase suicide gene therapy using three-dimensional cell cultures. AB - The efficacy of cancer gene therapy depends critically on "bystander effects" by which genetic modification of tumor cells results in killing of unmodified cells in the local microenvironment. In gene-dependent enzyme-prodrug therapy, expression of a prodrug-activating suicide gene is used to generate a cytotoxic metabolite that diffuses to nontransduced cells. The objective of this study was to develop a physiologically relevant tissue culture model for quantifying bystander effects and to validate the model using as an example the activation of dinitrobenzamide prodrugs (e.g., CB 1954) by Escherichia coli aerobic nitroreductase (NTR). Bystander effects were measured in three-dimensional multilayer cocultures of NTR+ and NTR- cells by determining clonogenic survival curves for both cell types using V79, Skov3, or WiDr as parental cell lines. Bystander killing by CB 1954 was much more efficient in multilayers than monolayers at equivalent cell:medium ratios, whereas the chloromustard analogue of CB 1954 showed even greater efficiency. For a series of dinitrobenzamides, bystander killing in multilayers showed a positive correlation with prodrug lipophilicity and also correlated with the bystander effect in mixed tumor xenografts grown from the same NTR+ and NTR- WiDr cell lines (r(2) = 0.84; P < 0.001). The multilayer model identified a bromomustard prodrug (SN 24927) with superior therapeutic activity to CB 1954 that provided curative activity against WiDr tumors comprising 1:1 mixtures of NTR+ and NTR- cells. This study demonstrates the utility of the multilayer tissue culture model for quantifying and optimizing bystander effects in tumors and identifies a new lead prodrug for NTR gene-dependent enzyme-prodrug therapy. PMID- 11888916 TI - Extracellular Bad fused to toxin transport domains induces apoptosis. AB - Bad, a proapoptotic member of the Bcl-2 family, is inactivated by phosphorylation, and this loss of activity may contribute to the malignancy of certain types of tumors such as glioblastoma and prostate cancer. To determine whether extracellular Bad can be delivered into cells via cell surface receptor binding and induce apoptosis, we genetically fused the mouse Bad gene to the gene for the translocation and receptor-binding domains of diphtheria toxin (DTTR). The purified Bad (wild-type)-DTTR protein showed cytotoxicity to human glioma cells in a dose-dependent manner. Bad phosphorylation sites at codons 112 and 136 were mutated from serine to alanine to prevent Bad inactivation by kinases and to increase the toxicity of Bad. The Bad (S112A S136A)-DTTR protein was at least 5 times more toxic than Bad (wild-type)-DTTR with an IC(50) of 5 x 10(-8) M. The Bad (S112A S136A)-DTTR protein altered the subcellular distribution of Bcl-X(L), indicating that it enters the cell cytoplasm and binds Bcl-X(L). Bad (S112D S136A)-DTTR, mutated to mimic phosphorylation of Bad, showed lower toxicity than either Bad (wild-type)-DTTR or Bad (S112A S136A)-DTTR, additionally indicating that Bad-DTTR must bind Bcl-X(L) to stimulate apoptosis. We conclude that extracellular Bad can be delivered into cells via the transport domain of a bacterial toxin and may be used to induce apoptosis. PMID- 11888917 TI - Effects of SP500263, a novel, potent antiestrogen, on breast cancer cells and in xenograft models. AB - We have compared the antitumor activities of SP500263, a novel next-generation selective estrogen receptor modulator (SERM), tamoxifen, and raloxifene side-by side in in vitro and in vivo MCF-7 breast cancer models. In vitro, SP500263 acted as an antiestrogen and potently inhibited estrogen-dependent MCF-7 proliferation with IC(50) values in the nanomolar range. SP500263 also strongly inhibited MCF-7 proliferation in the absence of estrogen at all of the concentrations tested. To investigate the antitumor activity of SP500263 in animals, athymic nude mice were implanted with MCF-7 tumor in the presence of a tumor growth-supporting sustained release estrogen pellet. Treatment was initiated after tumors were established. SP500263, administered for 28 days through daily i.p. dosing, effectively reduced estrogen-stimulated tumor growth at 3 and 30 mg/kg. SP500263 was as efficacious as tamoxifen and superior to raloxifene at the corresponding doses. Maximum efficacy was reached with the 30 mg/kg dose. The observed effects were highly significant. SP500263 represents a member of a novel series of SERMs that is structurally unrelated to SERMs currently on the market or in clinical development. The experiments described herein demonstrate that SP500263 is efficacious in the MCF-7 proliferation assay and in a murine model of breast cancer. PMID- 11888918 TI - Transformed and tumor-derived human cells exhibit preferential sensitivity to the thiol antioxidants, N-acetyl cysteine and penicillamine. AB - Thiol antioxidants, typified by N-acetyl cysteine, are known to induce p53 dependent apoptosis in transformed mouse embryo fibroblasts but not in normal mouse embryo fibroblasts. We now report that this is also the case for human cells. First, we used an isogenic fibroblast cell lineage exhibiting progressive stages of transformation, from primary derived cells to v-MYC immortalized to tumorigenic. At the immortalization stage, cells became 12- and 480-fold more sensitive to the thiol antioxidants N-acetyl cysteine (NAC) and penicillamine (PEN), respectively. Although immortalization of these cells was associated with v-MYC expression, overexpression of MYC was not sufficient for sensitizing these cells to antioxidants. To test whether sensitivity to antioxidants is a general property of immortalized human cells, including fully transformed cells, 12 tumor derived cell lines were treated with PEN, the more potent of the two antioxidants. Ten of 11 caspase-proficient tumor cell lines underwent apoptosis after treatment, whereas primary fibroblasts and keratinocytes were resistant. The difference between normal and transformed cells was apparent whether the assay used measured caspase 3 activation, Annexin V binding, or cell viability. Tumor cell lines containing wild-type p53 were more sensitive than p53-null cell lines. The requirement for p53 was tested using the p53 inhibitor, pifithrin alpha, or using stable transfectants of a v-MYC-immortalized, telomerase-positive cell line that expresses HPV16 E6 to bind and degrade p53. In the latter case, > or = 80% of the PEN-induced apoptosis was dependent on the presence of wild-type p53. These studies suggest that treatment with thiol-containing antioxidants, such as PEN, may offer a useful approach for preferential induction of apoptosis in preneoplastic and neoplastic cells. PMID- 11888919 TI - Tumor cell apoptosis by irradiation-induced nitric oxide production in vascular endothelium. AB - We examined the effects of X-ray irradiation on endothelial nitric oxide (NO) production in bovine aortic endothelial cells (BAECs). Single irradiation of up to 60 Gy did not affect the expression of endothelial NO synthase (eNOS) mRNA, as assessed by reverse transcription-PCR, in BAECs. The basal level of intracellular Ca(2+) concentration and Ca(2+) mobilization induced by thapsigargin and ATP were also not affected by single irradiation. However, eNOS-mediated, Ca(2+)-dependent constitutional NO production could not be examined directly because irradiated BAECs showed continuous NO production, as measured by diaminofluorescein-2 fluorescence, without agonist stimulation. Expression of inducible NO synthase (iNOS) mRNA and protein was markedly increased by 2 Gy of irradiation, thereby indicating that the basal and continuous NO production in irradiated BAECs was due to the expression of iNOS. Hepatoma HepG2 cells cocultured with irradiated BAECs showed apoptosis in the presence of L-arginine, and the apoptosis was prevented by L-NAME (N(omega)-nitro-L-arginine methyl ester). These results indicate that single irradiation does not affect Ca(2+) mobilization and eNOS expression but induces the expression of iNOS in BAECs, and the latter property would be beneficial to induce apoptosis in the adjacent tumor cells. PMID- 11888920 TI - Enhanced photodynamic therapy efficacy with inducible suicide gene therapy controlled by the grp promoter. AB - Photodynamic therapy (PDT) is a promising cancer treatment involving the administration of a tumor-localizing photosensitizer followed by the photochemical generation of cytotoxic singlet oxygen. PDT elicits strong transcriptional activation of a variety of genes including stress response genes belonging to the glucose-regulated protein (grp) family. Oxidative stress and hypoxia can activate GRP-78, and both of these physiological insults occur in treated tissue during and/or after PDT. In the current study, we evaluated the grp promoter as a PDT-inducible molecular switch for controlled expression of the herpes simplex virus-thymidine kinase (HSV-tk) suicide gene in mouse mammary adenocarcinoma (TSA) cells and tumors stably transduced with the G1NaGrpTk retroviral expression vector. We also examined whether PDT-inducible expression of HSV-tk, together with systemic administration of ganciclovir, could enhance the tumoricidal responsiveness of PDT. Inducible expression of HSV-tk was observed after PDT in stably transduced TSA cells grown in culture and in TSA tumors growing in BALB/c mice. We also observed enhanced tumoricidal activity in mice with TSA tumors containing the G1NaGrpTk expression vector treated with PDT plus ganciclovir when compared with either treatment alone. Our results confirm that the grp promoter was able to effectively function as a molecular switch for the inducible expression of the HSV-tk gene after exposure to PDT. PMID- 11888921 TI - Combination of T-cell therapy and trigger of inflammation induces remodeling of the vasculature and tumor eradication. AB - In a transgenic mouse model of multistep carcinogenesis, highly angiogenic insulinomas contain an irregular vascular network and develop an intrinsic resistance to leukocyte infiltration and effector function. Even persistently high levels of activated tumor-specific T lymphocytes fail to eradicate the tumors. In contrast, we show that irradiation before adoptive transfer results in complete macroscopic tumor regression. Thus, effective tumor therapy requires a proinflammatory microenvironment that permits T cells to extravasate and to destroy the tumor. Early after initiation of the irradiation/adoptive transfer therapy, the capillary network reacquires an almost normal appearance, a likely consequence of strong induction of the chemokines monokine induced by IFN-gamma (Mig) and IFN-inducible protein 10 (IP10). This remodeling of the vasculature in a proinflammatory environment may directly affect lymphocyte extravasation and effector function. Therefore, irradiation/adoptive transfer therapy combines antigen-driven tumor cell eradication with anti-angiogenic effects on tumor endothelium, a powerful synergy that has not been previously appreciated. PMID- 11888922 TI - Cellular processing of a multibranched lysine core with tumor antigen peptides and presentation of peptide epitopes recognized by cytotoxic T lymphocytes on antigen-presenting cells. AB - We showed that pRL1a multiple antigen peptide (MAP)-sensitized dendritic cell (DC) and P815 cell lysis by pRL1a-specific B-24 CTL was blocked by incubating target cells at 4 degrees C during sensitization. The finding suggested that pRL1a MAP was mostly internalized in DC and P815 cells and produced pRL1a peptide epitopes for presentation with H-2L(d). Furthermore, we showed that sensitization with pRL1a MAP was inhibited by the addition of chloroquine, cycloheximide, and brefeldin A to the culture, but not by the addition of inhibitors for lysosomal proteases or proteasome. Inhibition of sensitization by the addition of chloroquine to the culture suggested the requirement of acidification of the endosomal compartment for pRL1a MAP processing. Inhibition of sensitization by the addition of cycloheximide and brefeldin A to the culture indicated the requirement of newly generated MHC class I antigen molecules and the involvement of transport of the peptide MHC class I complex from the endoplasmic reticulum to the Golgi. The findings suggested that pRL1a MAP in the endosomal compartment leaked to the cytosol, and degraded, and the pRL1a peptide produced was presented by the MHC class I pathway. PMID- 11888923 TI - Vaccination with poly-L-arginine as immunostimulant for peptide vaccines: induction of potent and long-lasting T-cell responses against cancer antigens. AB - Vaccines that induce high numbers of sustained T cell responses are urgently needed for the treatment of numerous diseases including cancer. Antigen presenting cells (APCs), the most important of which are dendritic cells, orchestrate antigen-dependent T cell responses in that they present antigens to T cells in an appropriate environment. Here we present evidence that after vaccination with a simple mixture of the cationic poly-amino acid poly-L-arginine and tumor antigen-derived peptide antigens, large numbers of antigen-specific T cells are induced and APCs mediate the generation of T lymphocytes. We observe that after s.c. injection, MHC class II(+) cells infiltrate injection sites and are loaded with large amounts of antigen in vivo under the influence of poly-L arginine. Consequently, numerous antigen-charged APCs can be detected in draining lymph nodes of vaccinated animals. Antigen-specific T cell responses induced are systemic and were readily detected more than 4 months after the last vaccination, the latest time point we measured. By contrast, even after repeat injections, we were consistently unable to detect antibody responses against poly-L-arginine, allowing this compound to be used for numerous booster injections. Clinical trials in cancer patients using poly-L-arginine as immunostimulant will be carried out in the near future. PMID- 11888924 TI - Molecular-cytogenetic analysis of HER-2/neu gene in BRCA1-associated breast cancers. AB - The BRCA1 tumor suppressor gene and the HER-2/neu oncogene are located in close proximity on the long arm of chromosome 17 (17q11-21). Absence of BRCA1 or functional overexpression of the HER-2/neu gene presumably contributes to the somatic phenotype of breast cancer in premenopausal women, characterized by unfavorable prognostic features such as high tumor grade, hormone receptor negativity, and high proliferation rate. To examine whether amplification of HER 2/neu contributes to the aggressive biology of BRCA1-associated tumors, we have performed fluorescence in situ hybridization on formalin-fixed paraffin-embedded breast tumor tissue sections from 53 BRCA1 mutation carriers and 41 randomly selected, age-matched sporadic breast cancer cases. Although BRCA1-associated and sporadic tumors were equally likely (19% versus 22%) to exhibit HER-2/neu amplification [defined as a ratio of HER-2/neu copies to chromosome 17 centromere (CEP17) copies > or = 2], 6 (15%) of the sporadic tumors were highly amplified (defined as a ratio greater-than-or-equal 5) versus none of the BRCA1-associated tumors (P = 0.048). HER-2 protein overexpression as measured by immunohistochemical analysis was not observed among the BRCA1-associated cases (P = 0.042). Four of 21 (19%) sporadic tumors exhibited strong membranous staining of HER-2 (intensity level of 3+) as compared with 0 of 39 BRCA1-associated tumors. Our data suggest that a germ-line mutation in the BRCA1 tumor suppressor gene is associated with a significantly lower level of HER-2/neu amplification. Thus, it is possible that BRCA1-associated and HER-2/neu-highly amplified tumors progress through distinct molecular pathways, and the aggressive pathological features of BRCA1-associated tumors appear unrelated to amplification of the adjacent HER-2/neu oncogene. PMID- 11888925 TI - A new player in oncogenesis: AUF1/hnRNPD overexpression leads to tumorigenesis in transgenic mice. AB - AUF1/heterogeneous nuclear ribonucleoprotein D (hnRNPD) binds to adenylate uridylate-rich elements contained in the 3' untranslated region of many short lived mRNAs. This binding has been shown in vitro to control the stability of adenylate uridylate-rich element-containing mRNAs, including mRNAs encoding proto oncogenes, cytokines, or other signaling molecules. However, no studies have yet been undertaken to identify the mRNAs subject to AUF1-mediated regulation in vivo. The purpose of our study was to investigate the biological functions of AUF1. Thus, we derived transgenic (Tg) mice, which overexpress one isoform of AUF1, the p37(AUF1). Mice of the three Tg lines analyzed exhibit altered levels of expression of several target mRNAs, such as c-myc, c-jun, c-fos, granulocyte macrophage colony-stimulating factor, and tumor necrosis factor alpha. The Tg line with the highest amount of Tg p37(AUF1) protein developed sarcomas. The tumors strongly expressed AUF1 Tg protein and Cyclin D1. Taken together, our data show that: (a) AUF1 is a key regulatory factor of gene expression in vivo; and (b) the deregulation of this heterogeneous nuclear ribonucleoprotein leads to tumorigenesis. PMID- 11888926 TI - Functional analysis of 44 mutant androgen receptors from human prostate cancer. AB - Mutations of the androgen receptor gene are believed to contribute to the androgen-independent growth of prostate cancer cells. To date, 56 missense mutations of the androgen receptor have been identified in human prostate cancer. The functional status of most of these mutants has not yet been investigated. To address their functional properties, we generated 44 androgen receptor mutants that have been identified in human prostate cancer and used a colorimetric yeast reporter assay to analyze their transactivational activities in response to seven different ligands. We found that these mutant androgen receptors exhibited diverse transactivational activity: seven (16%) showed loss of function, three (7%) had wild-type function, 14 (32%) had partial function, and 20 (45%) had gains of function. Five of 20 gain-of-function mutants had promiscuous activity, being transactivated by non-androgens. We also found that the combination of estradiol and progesterone at physiological concentrations weakly or moderately activated an additional seven mutant androgen receptors. Our findings provide essential information for understanding the role of mutant androgen receptors in prostate cancer. PMID- 11888927 TI - Aberrant splicing in several human tumors in the tumor suppressor genes neurofibromatosis type 1, neurofibromatosis type 2, and tuberous sclerosis 2. AB - Mutations at splice sites or surrounding sequences have been reported to cause aberrant splicing. However, splicing errors can also occur without sequence alterations. We investigated three tumor suppressor genes for aberrant splicing in tumors. At a low frequency per exon it was found in five of seven of the investigated in-frame exons of the neurofibromatosis type 1 (NF1) gene, in two of three exons of the neurofibromatosis type 2 (NF2) gene, and in one of three exons of the tuberous sclerosis 2 gene. It was detectable in all of the human tumor tissues tested (NF1 neurofibroma, sporadic intramedullar neurinoma, sporadic meningiomas, NF2 schwannoma, NF2 meningioma, basalioma, and naevus) as well as in cultured tumor cell lines and cultured primary cells. Hence, our data show that aberrant splicing is a very common process. According to simulations of the secondary structures of the pre-mRNA, we suggest that aberrant splicing is attributable to the rare occurrence of alternative structures at the splice donor site, which are not recognized by the splice machinery. In HeLa cells, aberrant splicing is found to be increased at elevated temperatures and low pH in vitro, conditions often found in tumor tissues. In three tumor tissues tested for one NF1 exon, we found approximately twice the amount of aberrant transcript as in normal tissues. Therefore, we suggest that the increase in aberrant splicing caused by environmental factors represents an additional mechanism for the reduction of the amount of tumor suppressor mRNA in the absence of relevant mutations in the tumor. PMID- 11888928 TI - The tumorigenic diversity of the three PLAG family members is associated with different DNA binding capacities. AB - Pleomorphic adenoma gene (PLAG) 1, the main translocation target in pleomorphic adenomas of the salivary glands, is a member of a new subfamily of zinc finger proteins comprising the tumor suppressor candidate PLAG-like1 (also called ZAC1 or lost on transformation 1) and PLAGL2. In this report, we show that NIH3T3 cells overexpressing PLAG1 or PLAGL2 display the typical markers of neoplastic transformation: (a) the cells lose cell-cell contact inhibition; (b) show anchorage-independent growth; and (c) are able to induce tumors in nude mice. In contrast, PLAGL1 has been shown to prevent the proliferation of tumor cells by inducing cell cycle arrest and apoptosis. This difference in function is also reflected in their DNA binding, as we show here that the three PLAG proteins, although highly homologous in their DNA-binding domain, bind different DNA sequences in a distinct fashion. Interestingly, the PLAG1- and PLAGL2-induced transformation is accompanied by a drastic up-regulation of insulin-like growth factor-II, which we prove is a target of PLAG1 and PLAGL2. This strongly suggests that the oncogenic capacity of PLAG1 and PLAGL2 is mediated at least partly by activating the insulin-like growth factor-II mitogenic pathway. PMID- 11888929 TI - Tissue-specific deletion and discontinuous loss of heterozygosity are signatures for the mutagenic effects of ionizing radiation in solid tissues. AB - The mouse Aprt locus on chromosome 8 was used as the selectable target for the study of spontaneous and ionizing radiation-induced mutations in kidney epithelia and ear fibroblasts. Fifty-two Aprt heterozygous mice were exposed to 7.5 Gy of (137)Cs-gamma radiation on their right sides, and Aprt-deficient clones were isolated from enzymatically digested tissues at times ranging from 1 day to 14 months after irradiation. A statistically significant increase in the mutant frequencies for the irradiated tissues was observed when compared with the spontaneous mutant frequencies for the nonirradiated tissues. A molecular analysis of spontaneous mutations observed for the nonirradiated tissues revealed tissue-specific differences; apparent chromosome loss was common in kidney mutants but infrequent in the ear mutants, whereas apparent deletions were common in the ear mutants but not detected in the kidney mutants. For the irradiated kidneys, apparent deletions were observed commonly demonstrating that these events are markers for ionizing radiation mutagenesis in this tissue. All of the loss of heterozygosity (LOH) tracts observed in the spontaneous mutants were continuous, but discontinuous LOH patterns were observed in 6--8% of ionizing radiation-induced ear and kidney cell mutants. Work with kidney-derived cell lines showed that discontinuous LOH is a novel signature for delayed ionizing radiation mutagenesis. Considered together, these results suggest that ionizing radiation-induced mutations in vivo can result from both direct and delayed mutagenic effects. PMID- 11888930 TI - Deficiency in DNA polymerase beta provokes replication-dependent apoptosis via DNA breakage, Bcl-2 decline and caspase-3/9 activation. AB - Cells deficient in DNA polymerase beta (beta-pol) are impaired in base excision repair (BER) and hypersensitive to various DNA damaging agents, including methylating mutagens. Hypersensitivity of beta-pol-deficient cells to methylating agents is because of induction of apoptosis (Ochs et al., Cancer Res., 59: 1544 1551, 1999), indicating incompletely repaired DNA damage to trigger the response. Here we show that defective BER in beta-pol-null cells results in an early and transient increase in the frequency of DNA single-strand breaks on treatment with methyl methanesulfonate. These breaks arising as repair intermediates are not likely to trigger apoptosis directly because they were repaired efficiently and generated both in resting and proliferating cells, whereas only proliferating cells underwent with high frequency apoptosis after methylation. Therefore, we propose that single-strand breaks are converted into another kind of critical apoptosis-triggering lesion during replication. These critical secondary DNA lesions are likely to be non-repaired DNA double-strand breaks (DSBs), which are formed at higher frequency in beta-pol-null than in wild-type cells. Apoptosis was a late response not detectable before 24 h after methylation and was preceded by DSBs formation, extensive chromosomal breakage, and decline in Bcl-2 level and caspase-9 and caspase-3 activation. Caspase-8 was not significantly activated. Transfection of beta-pol-null cells with bcl-2 protected against methylation induced apoptosis, indicating Bcl-2 to be causally involved. Overall, the data demonstrate that in cells lacking beta-pol, defective BER results in incompletely repaired DNA damage, which triggers apoptosis in a replication-dependent way by activating the mitochondrial death pathway. It is suggested that DSBs act as a critical ultimate apoptosis-inducing lesion. PMID- 11888931 TI - Allelic imbalance of 7q32.3-q36.1 during tumorigenesis in Barrett's esophagus. AB - Malignant transformation of Barrett's esophagus is characterized by three distinct premalignant stages: intestinal metaplasia (MET), low- (LGD), and high grade dysplasia (HGD). We reported recently an increase in the frequency of loss of 7q33-q35 between LGD and HGD as determined by comparative genomic hybridization (P. H. J. Riegman et al., Cancer Res., 61: 3164-3170, 2001). Now the 7q32.3-q36.1 region was additionally characterized by allelotype analysis with 11 polymorphic markers in 15 METs, 20 LGDs, 20 HGDs, and 20 Barrett's adenocarcinomas from different patients. Low percentages of imbalance were determined in METs and LGDs, 7% and 10%, respectively, whereas HGDs and Barrett's adenocarcinomas revealed high percentages of loss, 75% and 65%, respectively. This difference in frequency between LGDs and HGDs appeared highly significant: P = 0.00007. The majority of imbalances were found at D7S2439 and D7S483, located on 7q36.1. These data suggest that markers from this area can be used as a diagnostic tool in Barrett's esophagus, i.e., to distinguish between watchful waiting and active treatment. PMID- 11888932 TI - Real-time optical imaging of primary tumor growth and multiple metastatic events in a pancreatic cancer orthotopic model. AB - We report here whole-body optical imaging, in real time, of genetically fluorescent pancreatic tumors growing and metastasizing to multiple sites in live mice. The whole-body optical imaging system is external and noninvasive. Human pancreatic tumor cell lines, BxPC-3 and MiaPaCa-2, were engineered to stably express high-levels of the Aequorea victoria green fluorescent protein (GFP). The GFP-expressing pancreatic tumor cell lines were surgically orthotopically implanted as tissue fragments in the body of the pancreas of nude mice. Whole body optical images visualized real-time primary tumor growth and formation of metastatic lesions that developed in the spleen, bowel, portal lymph nodes, omentum, and liver. Intravital images in the opened animal confirmed the identity of whole-body images. The whole-body images were used for real-time, quantitative measurement of tumor growth in each of these organs. Intravital imaging was used for quantification of growth of micrometastasis on the liver and stomach. Whole body imaging was carried out with either a trans-illuminated epi-fluorescence microscope or a fluorescence light box, both with a thermoelectrically cooled color CCD camera. The simple, noninvasive, and highly selective imaging made possible by the strong GFP fluorescence allowed detailed simultaneous quantitative imaging of tumor growth and multiple metastasis formation of pancreatic cancer. The GFP imaging affords unprecedented continuous visual monitoring of malignant growth and spread within intact animals without the need for anesthesia, substrate injection, contrast agents, or restraint of animals required by other imaging methods. The GFP imaging technology presented in this report will facilitate studies of modulators of pancreatic cancer growth, including inhibition by potential chemotherapeutic agents. PMID- 11888933 TI - Identification of heat shock protein 60 as a molecular mediator of alpha 3 beta 1 integrin activation. AB - The alpha 3 beta 1 integrin is involved in the adhesion of metastatic breast cancer cells to the lymph nodes and to osteoblasts in the bone. Regulation of the affinity or avidity of integrins for their ligands may result from conformational changes induced by changes in the microenvironment of the integrin. Two surface proteins, 55 and 32 kDa, coimmunoprecipitated with the alpha 3 beta 1 integrin from breast carcinoma cells. The 55-kDa protein preferentially associated with the active form of the alpha 3 beta 1 integrin. The protein was identified as HSP60 using two-dimensional electrophoresis and mass spectrometry and confirmed by reimmunoprecipitation of the integrin immune complex with an anti-HSP60 antibody. In cell spreading assays on a thrombospondin-1 substrate, addition of exogenous-recombinant HSP60 was sufficient to specifically activate alpha 3 beta 1 integrin but not to activate function of alpha 2 beta 1, alpha v beta 3, alpha 4 beta 1, or alpha 5 beta 1 integrins. Furthermore, mizoribine, an HSP60-binding drug, blocked activation of the alpha 3 beta 1 integrin induced by insulin-like growth factor 1 (IGF1) or exogenous recombinant HSP60 and inhibited the association of HSP60 with the integrin. Additionally, inhibiting the surface expression of endogenous HSP60 by nonactin inhibited activation of the alpha 3 beta 1 integrin by IGF1. These data demonstrate that HSP60 binding is sufficient to activate alpha 3 beta 1 integrin function and suggest that association of endogenous HSP60 with alpha 3 beta 1 integrin is necessary for IGF1-induced activation. PMID- 11888934 TI - Activation of the Erk mitogen-activated protein kinase pathway stimulates neuroendocrine differentiation in LNCaP cells independently of cell cycle withdrawal and STAT3 phosphorylation. AB - Neuroendocrine (NE) differentiation in prostate cancer (PCa) has been found in some studies to correlate with unfavorable clinical outcome. The mechanisms by which PCa acquires NE properties are poorly understood. In this study, we demonstrate that heparin-binding epidermal growth factor-like growth factor (HB EGF), a prostate smooth muscle-derived mitogen and survival factor, can evoke NE differentiation in LNCaP human PCa cells. HB-EGF induction of NE differentiation was mediated by a mitogen-activated protein kinase (MAPK) kinase-dependent mechanism, and this process was blocked by p38 MAPK signaling. NE differentiation induced by HB-EGF occurred independently of STAT3 phosphorylation and coincided with continued cell cycle transit. These findings suggest that endogenous stroma derived factors, acting through MAPK signaling pathways, may play a significant role in the acquisition of NE properties by PCa cells. They also demonstrate that withdrawal from the cell cycle is not a prerequisite for expression of NE characteristics by PCa. PMID- 11888935 TI - Improved magnetic resonance imaging detection of prostate cancer in a transgenic mouse model. AB - Transgenic mouse models of prostate cancer provide an opportunity to conduct genetic tests of the molecular mechanisms underlying initiation and progression of tumorigenesis. They also allow assessment of the effects of various pharmacological interventions. However, one limitation that has impeded full exploitation of these models is the lack of in vivo imaging procedures of sufficient sensitivity and resolution to detect and follow tumors at early stages of growth. We have addressed this problem through the use of diffusion-weighted magnetic resonance imaging (DWI). A transgenic mouse model (CR2-TAg) of prostate cancer was used to show that DWI can detect tumors <1 mm in diameter. Markedly enhanced DWI contrast results from a 2-fold difference in apparent diffusion coefficient between benign and malignant prostatic tissue (P < 0.00001). Clinical application of DWI may offer advantages over current T2-weighted magnetic resonance imaging methods. PMID- 11888936 TI - Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), a novel Hsp90-client tyrosine kinase: down-regulation of NPM-ALK expression and tyrosine phosphorylation in ALK(+) CD30(+) lymphoma cells by the Hsp90 antagonist 17-allylamino,17 demethoxygeldanamycin. AB - Anaplastic large cell lymphomas (ALCL) are characterized by the expression of a chimeric protein, NPM-ALK, which originates from fusion of the nucleophosmin (NPM) and the membrane receptor anaplastic lymphoma kinase (ALK) genes. The NPM ALK kinase, on dimerization, shows phosphotransferase activity and, through its interaction with various ALK-adapter proteins, induces cell transformation and increases cell proliferation in vitro. The chaperones heat shock proteins 90 (Hsp90) and 70 (Hsp70) play a critical role in the folding and maturation of several oncogenic protein kinases, and perturbation of Hsp90 structure affects the stability and degradation of Hsp90- and Hsp70-bound substrates. This process is triggered by benzoquinone ansamycin antibiotics, Hsp90-binding small molecules. We have studied the effect of 17-allylamino,17-demethoxygeldanamycin (17-AAG), a benzoquinone ansamycin, on NPM-ALK steady-state level in ALCL cells. Treatment with 17-AAG decreased NPM-ALK expression and phosphorylation, thus impairing its association with phospholipase C-gamma, Src homology 2 domain containing protein (Shc), growth factor receptor-bound protein 2 (Grb2), and insulin receptor substrate-1 (IRS-1). We also observed that NPM-ALK associates with Hsp90, and incubation with 17-AAG disrupts this complex without affecting Hsp90 expression. As shown previously for other Hsp90 client proteins, destabilization of the Hsp90/NPM-ALK complex induced by 17-AAG resulted in increased binding of the chimeric protein to Hsp70, which is known to affect protein degradation. Hsp/NPM-ALK complex formation appears to be independent of NPM sequences, because we were unable to coimmunoprecipitate NPM with either Hsp90 or Hsp70. Similar to NPM-ALK, the exogenously expressed variant fusion protein TPR-ALK showed decreased expression and phosphorylation after 17-AAG treatment, suggesting that the effect of 17-AAG on ALK chimeric proteins depends on the ALK portion and not on the partner protein moiety. Our data demonstrate that NPM-ALK cell content is determined by its interaction with Hsp90 and Hsp70, and suggest that the alteration of such associations can interfere with NPM-ALK function in ALCL cells. PMID- 11888937 TI - Cyclooxygenase-2 activity altered the cell-surface carbohydrate antigens on colon cancer cells and enhanced liver metastasis. AB - Cyclooxygenase-2 (COX-2) was recently reported (M. Tsujii and R. N. DuBois, Cell, 83: 493-501, 1995) to affect the metastatic potential of cells. Previous studies (M. Fukuda, Cancer Res., 56: 2237-2244, 1996) indicated that sialyl Lewis antigen expression is correlated with hematogenous metastasis of colon cancer. In the present study, we investigated the interaction between COX-2 activity, expression of sialyl Lewis antigens, in vitro cancer cell adhesion to endothelial cells, and in vivo metastatic potential. Effects of COX-2 activity and prostaglandin E(2) on cell adhesion, expression of sialyl Lewis antigens, and glycosyltransferase genes were determined in Caco-2-m (COX-2 low level), Caco-2-COX-2 (programmed to overexpress COX-2), and HT-29 (COX-2 high level) cells. Metastatic spread of these cells to the liver was also investigated. Caco-2-COX-2 cells had increased SPan-1 levels and increased adherence to endothelial cells via SPan-1 compared with Caco-2-m cells. HT-29 cells expressed sialyl Lewis a and adhered to endothelial cells via sialyl Lewis a. Treatment with a COX-2 inhibitor, celecoxib, decreased SPan-1 and sialyl Lewis a expression and adherence to endothelial cells. beta 3Gal-T5 and ST3Gal III and IV expression was inhibited by celecoxib and was enhanced by prostaglandin E(2) treatment. Caco-2-COX-2 and HT 29 cells metastasized to the liver, whereas Caco-2-m cells did not. Pretreatment with celecoxib reduced the metastatic potential as well as anti-sialyl Lewis antibodies. Our results indicate a direct link between COX-2 and enhanced adhesion of carcinoma cells to endothelial cells, and enhanced liver metastatic potential via accelerated production of sialyl Lewis antigens. COX-2 inhibitors may suppress metastasis. PMID- 11888939 TI - Is asthma another interstitial lung disease? PMID- 11888940 TI - Treatment of obstructive sleep apnea: no longer just a lot of hot air. PMID- 11888941 TI - Pulmonary blastomycosis: a great masquerader. PMID- 11888942 TI - Prevention of ventilator-associated pneumonia: selecting interventions that make a difference. PMID- 11888943 TI - Is a silver coating a silver lining? PMID- 11888944 TI - End-of-life issues and the do-not-resuscitate order: who gives the order and what influences the decision? PMID- 11888945 TI - Tobacco dependence: a chronic disease. PMID- 11888946 TI - Measurement of short-term changes in dyspnea and disease-specific quality of life following an acute COPD exacerbation. AB - STUDY OBJECTIVE: To determine whether currently available measurement tools can be used to obtain valid measurements of short-term changes in dyspnea and disease specific quality of life (QOL) in outpatients with an acute COPD exacerbation. DESIGN: Prospective cohort study. METHODS: Sixty-six patients with an acute COPD exacerbation who presented to the emergency department completed the chronic respiratory disease index questionnaire (CRQ) and the baseline dyspnea index (BDI) and were discharged home receiving 10 days of medical therapy. Reassessment with the CRQ and the transitional dyspnea index (TDI) occurred within 48 h of relapse (defined as an urgent hospital revisit within 10 days because of worsening respiratory symptoms), or 10 days later if relapse did not occur. RESULTS: Patients who did not relapse (n = 49) showed moderate-to-large improvements in disease-specific QOL across all four CRQ domains (improvements in each domain of 1.4 to 1.9 U; p < 0.001 for all domains) and large positive changes in the TDI (total TDI score, + 5.02 plus minus 0.55 U; p = 0.0001). In contrast, patients who had a relapse (n = 17) did not have improved CRQ or TDI scores (mean negative change in three of four CRQ domains, total TDI score - 3.06 plus minus 1.14 U; p = 0.02). Changes in the CRQ dyspnea score and TDI correlated with each other (r = 0.78; p = 0.0001) and with changes in FEV(1) (CRQ, r = 0.48 and p = 0.0001; TDI, r = 0.46 and p = 0.0002). Ten control patients with stable COPD showed no changes in the CRQ or TDI over 10 days. CONCLUSION: The CRQ and BDI/TDI can be used to obtain valid, responsive measures of acute changes in QOL and dyspnea associated with a COPD exacerbation. The direction and magnitude of change in these scores was highly correlated with clinical outcome and with other health measures. Most outpatients treated for a COPD exacerbation experience significant short-term improvements in QOL and dyspnea, with the exception of patients who have a clinical relapse of symptoms. PMID- 11888948 TI - Association between the angiotensin-converting enzyme gene polymorphisms and tissue oxygenation during exercise in patients with COPD. AB - STUDY OBJECTIVES: We have recently determined that the angiotensin-converting enzyme (ACE) DD genotype might be associated with pulmonary hypertension during exercise in patients with COPD. Therefore, this study was designed to determine whether ACE gene polymorphisms adversely affect tissue oxygenation during exercise in patients with COPD. DESIGN: Cross-sectional analysis. SETTING: University hospital. PATIENTS: Thirty-nine patients (14 patients with II genotype, 12 patients with ID genotype, and 13 patients with DD genotype). INTERVENTIONS: All patients underwent right-heart catheterization and constant load exercise testing for 5 min on an ergometer. MEASUREMENTS AND RESULTS: The ratio of the change in oxygen delivery (DO(2)) to the increase in oxygen consumption (O(2)) during exercise (DeltaDO(2)/DeltaO(2)) was significantly lower in patients with the DD genotype (1.5 +/- 0.2) than in those with the II genotype (1.9 +/- 0.3, p = 0.0006) and the ID genotype (1.7 +/- 0.2, p = 0.037). Mixed venous oxygen tension (PO(2)) after exercise in patients with the DD genotype (23.5 +/- 1.5 mm Hg) was also significantly lower than in patients with the II genotype (26.7 +/- 1.6 mm Hg, p = 0.0002) and the ID genotype (25.0 plus minus 2.0 mm Hg, p = 0.045). In addition, the change in plasma concentration of lactate during exercise (DeltaLactate) was significantly higher in patients with DD genotype (33.3 +/- 4.3 mmol/L) than in those with the II genotype (25.5 +/- 3.6 mmol/L, p = 0.0002) and the ID genotype (28.8 +/- 4.0 mmol/L, p = 0.029). The mean pulmonary arterial pressure after exercise was significantly correlated with DeltaDO(2)/DeltaO(2) (r = - 0.423, p = 0.0076) but not with PvO(2) after exercise and with DeltaLactate. CONCLUSIONS: The ACE DD genotype may be associated with an impairment in peripheral tissue oxygenation during exercise in patients with COPD. PMID- 11888949 TI - Use of a mucus clearance device enhances the bronchodilator response in patients with stable COPD. AB - STUDY OBJECTIVE: To determine whether the use of a mucus clearance device (MCD) [Flutter; Axcan Scandipharm; Birmingham, AL] could improve the bronchodilator response to inhaled ipratropium and salbutamol delivered by a metered-dose inhaler in patients with stable, severe COPD. PATIENTS: Twenty-three patients with severe COPD were studied. Mean +/- SD age was 71.7 +/- 6.3 years. Mean FEV(1) was 0.74 +/- 0.28 L or 34.5 +/- 12.7% predicted. METHODS: Patients were tested in random order on 2 subsequent days after using an MCD or a sham MCD. A bronchodilator (four puffs; each puff delivering 20 microg of ipratropium bromide and 120 microg of salbutamol sulfate) was administered by metered-dose inhaler with a holding chamber after use of the MCD or sham MCD. Spirometry was performed before and after use of the MCD or sham MCD, and at 30 min, 60 min, and 120 min after the bronchodilator. Six-minute walk distance was tested between 30 min and 60 min; oxygen saturation, pulse, and a dyspnea score were recorded before and after walking. RESULTS: Immediately after use of the MCD, but not the sham MCD, there was a statistically significant (p < 0.05) improvement in FEV(1) and FVC (11 +/- 24% vs 1 +/- 7% and 18 +/- 33% vs 6 +/- 18%, respectively). Whether patients were pretreated with the MCD or sham MCD, there was a significant improvement in FEV(1) and FVC compared to baseline with combined bronchodilator therapy. At 120 min, the change in FEV(1) after treatment with the MCD was greater than with the sham MCD (186 +/- 110 mL vs 130 +/- 120 mL; p < 0.05). When comparing the MCD to the sham MCD, 6-min walk distance was greater (174 +/- 92 m vs 162 +/- 86 m; p < 0.05), with less dyspnea before and at the end of walking. CONCLUSION: Patients with severe COPD may demonstrate a significant bronchodilator response to combined ipratropium and salbutamol delivered by metered-dose inhaler. This response may be enhanced and additional functional improvement obtained with the prior use of a bronchial MCD. PMID- 11888950 TI - Repeatability of inspiratory capacity during incremental exercise in patients with severe COPD. AB - STUDY OBJECTIVE: Estimating lung volume using inspiratory capacity (IC) maneuvers is a useful way of tracking dynamic hyperinflation. An understanding of the repeatability of the IC in a clinical setting is important when evaluating an individual's response to a therapeutic intervention that might influence lung volume. This is the first study to determine the repeatability of serial IC measurements of patients with severe COPD undergoing incremental exercise testing in a clinical setting. SUBJECTS AND METHODS: Ten patients with severe COPD, inexperienced in exercise testing, cycled with power increased until they reached symptom limitation. Flow was measured at the mouth using a pneumotachograph. IC maneuvers were performed at 1-min to 3-min intervals. Subjects repeated the exercise test 2 days later. Three methods of calculating IC from flow have been described previously. To determine which method provided the best repeatability, we calculated the following: (1) IC calculated by the integration of inspired flow from the start to the end of the IC maneuver (ICINSP); (2) IC calculated from the difference between the drift-corrected peak inspiratory volume (total lung capacity [TLC]) and the drift-corrected end-expiratory lung volume (EELV) of the six breaths that preceded the IC prompt (ICREG); and (3) IC calculated, after correction of the expiratory part of the signal, as the difference between the mean EELV of the six breaths that preceded the IC prompt and the peak inspiratory volume (ICRATIO). Each individual's IC response was expressed as a function of exercise time and of ventilation. RESULTS: There was a significant (p < 0.05) decrease in the expired volume of the breath before the IC maneuver (0.11 +/- 0.26 L) [mean +/- SD]. ICINSP (1.78 +/- 0.88 L) was significantly less than the IC calculated using the other two methods (ICREG, 1.88 +/- 0.89 L; ICRATIO, 1.86 +/- 0.87 L). ICRATIO improved the repeatability of the serial IC measures by as much as 60% over ICINSP and ICREG. CONCLUSION: Calculating IC as the difference between EELV and TLC was unaffected by unsatisfactory technique, such as a change in breathing pattern immediately before the maneuver. Adjusting expiratory flow based on premaneuver inspiratory to expiratory volume ratio before estimating EELV improved the repeatability coefficient of the IC. PMID- 11888952 TI - Asthma, allergy, and airway hyperresponsiveness are not linked to the beta(2) adrenoceptor gene. AB - STUDY OBJECTIVES: To exclude genetic linkage between the beta(2)-adrenoceptor gene and asthma, allergy, and methacholine airway hyperresponsiveness. DESIGN: The current study used six distinct intragene markers within the beta(2) adrenoceptor gene, and evaluated genetic linkage between the beta(2)-adrenoceptor and asthma, allergy, or methacholine airway hyperresponsiveness in eight multiplex families. PATIENTS: Forty-nine members of eight multiplex families with a high incidence of asthma. INTERVENTIONS: Phenotypes were characterized by history, physical examination, skin testing, pulmonary function tests, and methacholine inhalational challenge. Genetic loci were identified using restriction fragment length polymorphisms, denaturing gradient gel electrophoresis, and restriction enzyme digest of polymerase chain reaction amplified fragments of the beta(2)-adrenoceptor gene. MEASUREMENTS AND RESULTS: Nonparametric analysis using computer analysis software found no evidence for linkage between these markers within the beta(2)-adrenoceptor gene and asthma. Parametric exclusion analysis using a dominant inheritance model resulted in large negative lod scores (- 6.74, - 19.44, and - 49.9, respectively) for tight linkage between asthma, allergy, or methacholine airway hyperresponsiveness and these polymorphic markers. CONCLUSIONS: These results indicate that asthma, allergy, and methacholine airway hyperresponsiveness are not linked to a dominant beta(2)-adrenoceptor gene with strong effect in these eight families with an inherited pattern of asthma. PMID- 11888951 TI - Unsuspected loss of lung elastic recoil in chronic persistent asthma. AB - STUDY OBJECTIVES: To investigate the progression and mechanism(s) for fixed maximum expiratory airflow (max) limitation in patients with chronic persistent asthma. METHODS: When optimally treated and in clinically stable condition, we studied 21 asthmatic patients and classified them into three groups based on the severity of expiratory airflow limitation: (1) group A included 5 asthmatic patients (four women; mean +/- SD age, 51 +/- 17 years) with mild persistent asthma (FEV(1) > 80% predicted) with serial FEV(1) measurements obtained prior to the present study for 16 +/- 4 years; (2) group B included 11 asthmatic patients (three women; mean age, 64 +/- 11 years) with moderate persistent asthma (FEV(1) of 60 to 80% predicted) with serial FEV(1) measurements for 12 +/- 4 years; and (3) group C included 5 asthmatic patients (three women; mean age, 55 +/- 16 years) with severe persistent asthma (FEV(1) < 60% predicted) with serial FEV(1) measurements for 11 plus minus 5 years. RESULTS: Lung CT indicated no or trivial emphysema, and diffusion was normal in all asthmatics. There was a marked loss of lung elastic recoil at total lung capacity (TLC) in all asthmatic patients in group B (16 +/- 4 cm H(2)O) and group C (15 +/- 5 cm H(2)O), but none or minimal in group A (22 +/- 1 cm H(2)O) [p < 0.01], and loss of elastic recoil accounted for 34% and 50% of decreased maximal expiratory airflow (max) at 80% and 70% TLC, respectively. Comparison with previous longitudinal data indicated individual asthmatics when in clinically stable condition remained predominantly in the same FEV(1) percent predicted classification group as in the current study. CONCLUSION: Patients with chronic moderate and severe persistent asthma, despite optimal therapy, have reduced max for many years in part due to (early?) loss of lung elastic recoil from unknown mechanism(s). This challenges current concept of airway remodeling. PMID- 11888953 TI - Reduction of eosinophilic inflammation in the airways of patients with asthma using montelukast. AB - OBJECTIVE: Leukotrienes (LTs) are involved in airway eosinophilic inflammation in patients with asthma. We examined the effects of a cysteinyl LT 1-receptor antagonist, montelukast, on sputum eosinophil levels, and the correlation between sputum eosinophils and bronchodilatation in patients with asthma. DESIGN: Double blind, randomized, crossover study. SETTING: University hospital and private hospital. PATIENTS: Twenty-nine patients with mild-to-moderate asthma. INTERVENTIONS: Montelukast, 10 mg, and placebo tablet, once daily, each for 4 weeks. MEASUREMENTS: Sputum eosinophils analyzed using hypertonic saline solution induced sputum and airway hyperresponsiveness to histamine were evaluated before and after treatment. In addition, morning and evening peak expiratory flow (PEF), asthma symptoms, and peripheral blood eosinophil levels were assessed. RESULTS: The percentage of eosinophils in sputum decreased from 24.6 +/- 12.3% at baseline to 15.1 +/- 11.8% after montelukast treatment, for a change of - 9.5 +/- 12.7% (n = 20). During placebo administration, the percentage of eosinophils fell from 21.3 +/- 12.1% to 21.0 +/- 11.5%, resulting in a decrease of - 0.3 +/- 10.8% (n = 20). There was a statistically significant difference in the change in sputum eosinophil levels between these two periods (p < 0.005). The number of peripheral blood eosinophils also significantly decreased after montelukast treatment (314.1 +/- 237.6/mL) compared with placebo (413.1 +/- 232.1/mL; p < 0.005, n = 21). Although morning and evening PEF values were significantly improved from baseline after montelukast treatment (p < 0.01, n = 20), asthma symptoms and airway responsiveness to histamine were not significantly altered. Furthermore, there was no significant correlation between the decrease in sputum eosinophils and the increase in PEF. CONCLUSION: These results suggest that montelukast has anti inflammatory effects on the airway in patients with asthma, and that its bronchodilatory effect is not solely dependent on a decrease in airway eosinophilia. PMID- 11888954 TI - 4-year follow-up of treatment with dental appliance or uvulopalatopharyngoplasty in patients with obstructive sleep apnea: a randomized study. AB - STUDY OBJECTIVES: To evaluate the effects of treatment with a dental appliance or uvulopalatopharyngoplasty (UPPP) on somnographic variables in patients with mild to-moderate obstructive sleep apnea (OSA) followed up for 4 years, and compliance and complementary treatment. DESIGN: Randomized study. SETTING: Central Hospital, Vasteras, Uppsala University, Sweden. PATIENTS: Ninety-five male patients with confirmed mild-to-moderate OSA (apnea index [AI] > 5 and < 25) were randomized to treatment with a dental appliance or UPPP. Sleep studies were performed before and 1 year and 4 years after intervention. Thirty-two patients in the dental appliance group and 40 patients in the UPPP group completed the 4-year follow-up. RESULTS: The success rate (percentage of patients with at least 50% reduction in AI) in the dental-appliance group was 81%, which was significantly higher than in the UPPP group, 53% (p < 0.05). Normalization (AI < 5 or apnea/hypopnea index < 10) was observed in 63% of the dental-appliance group and 33% of the UPPP group after 4 years. The difference between the groups was significant (p < 0.05). The compliance to use of the dental appliance was 62% at the 4-year follow-up. Thirty patients (75%) in the UPPP group continued without complementary treatment. The dental appliances had few adverse effects on the stomatognathic system, and the number of adjustments and repairs of the appliances over time was moderate. Pronounced complaints of nasopharyngeal regurgitation of fluid and difficulty with swallowing after UPPP were reported by 8% and 10%, respectively. CONCLUSIONS: The dental-appliance group showed significantly higher success and normalization rates regarding the somnographic variables compared to the UPPP group, but the effectiveness of the dental appliance was partly invalidated by the compliance of 62% at the 4-year follow-up. However, the appliances had few adverse effects on the stomatognathic system and required only moderate adjustments. Use of a dental appliance with regular follow-up can be recommended for long-term treatment of OSA. PMID- 11888955 TI - Prospective testing of two models based on clinical and oximetric variables for prediction of obstructive sleep apnea. AB - STUDY OBJECTIVE: To test the validity of two models for prediction of obstructive sleep apnea syndrome (OSAS) before polysomnography. DESIGN: Prospective study. SETTING: Sleep laboratory in an obesity clinic. PATIENTS: Data from two populations were analyzed: the first (group 1) included 102 consecutive overweight patients referred to our laboratory by an obesity clinic between May 1992 and November 1994, and was used to develop the prediction models. The second (group 2) included 108 consecutive new patients referred to our laboratory by the same obesity clinic between February 1997 and September 1998, and was used to test the prediction models. MEASUREMENTS AND RESULTS: Models were developed using a clinical score, pulmonary function tests, arterial blood gas tensions, and nocturnal pulse oximetry. OSAS was defined by an apnea-hypopnea index (AHI) > 15 events per hour, as measured by full-night polysomnography. Step-by-step multiple linear regression analysis (MLR) was used to provide an equation for calculation of predicted AHI, while logistic regression analysis (LR) provided an equation for calculation of the probability (P') of having OSAS. Characteristics of groups 1 and 2 were similar except for the prevalence of OSAS, which was higher in group 2 (74% vs 39% in group 1). The negative predictive value (NPV) of the MLR model dropped from 82.9% in group 1 to 36.7% in group 2. In parallel, the NPV of a P' < 0.25 according to LR decreased from 78.6% in group 1 to 23.5% in group 2. CONCLUSION: Our results emphasize the need for systematic prospective testing of mathematical predictive models in OSAS, since their diagnostic characteristics may differ markedly between populations, even when the setting and mode of recruitment remain unchanged. PMID- 11888956 TI - Home unattended vs hospital telemonitored polysomnography in suspected obstructive sleep apnea syndrome: a randomized crossover trial. AB - STUDY OBJECTIVE: To compare home unattended polysomnography (H-PSG) with polysomnography performed in a local hospital and telemonitored by a sleep laboratory (T-PSG) in the diagnosis of obstructive sleep apnea syndrome (OSAS). DESIGN: Randomized crossover trial. PATIENTS: Ninety-nine patients with suspected OSAS who underwent H-PSG and T-PSG on 2 consecutive nights, according to a randomized order. MEASUREMENTS: H-PSG and T-PSG were compared in terms of (1) effectiveness, only recordings providing interpretable signals from at least one EEG, the electro-oculograph, the electromyograph, air flow, thoracic or abdominal movements, and arterial oxygen saturation for 180 min of sleep were considered to be effective; (2) patient preference assessed by a questionnaire; and (3) polysomnographic indexes and final interpretative results in patients for whom both recordings were legible. RESULTS: Recordings were considered to be ineffective in 11.2% of T-PSG (95% confidence interval [CI], 4.9 to 17.4%) and in 23.4% of H-PSG (95% CI, 19.12 to 27.68%). Thermistor problems were the main cause of failure of H-PSG. Forty-one percent of patients preferred H-PSG, and 55% preferred T-PSG. H-PSG and T-PSG did not differ in terms of sleep and respiratory indexes in the 65 patients in whom both recordings were legible. H-PSG and T-PSG were concordant in 58 of 65 patients using a 10-event-per-hour apnea-hypopnea index cutoff value for the diagnosis of OSAS. CONCLUSIONS: T-PSG is clearly superior to H-PSG from a technical point of view and tends to be preferred by patients. The site of recording (home vs hospital) has no influence on polysomnographic indexes. PMID- 11888957 TI - Sleep structure correlates of continuous positive airway pressure variations during application of an autotitrating continuous positive airway pressure machine in patients with obstructive sleep apnea syndrome. AB - STUDY OBJECTIVES: To evaluate the relationship between sleep structure and continuous positive airway pressure (CPAP) delivered by an automatic CPAP (auto CPAP) machine in patients with obstructive sleep apnea syndrome (OSAS). DESIGN: Nocturnal polysomnography was performed during CPAP administration by an auto CPAP machine (Autoset Clinical 1; ResMed; Sydney, Australia). SETTING: Sleep disorders center in a research institute. PATIENTS: Fifteen subjects with newly diagnosed OSAS deserving home CPAP treatment. MEASUREMENTS AND RESULTS: During the night, in most cases, the lowest CPAP level was recorded during a prolonged nonrapid eye movement (NREM) sleep period uninterrupted by arousals, whereas the highest level during wake-sleep transitions or NREM sleep fragmented by arousals. In four subjects, rapid eye movement sleep was always associated with increasing CPAP. Sleep efficiency was negatively correlated with CPAP variability, evaluated as the SD of the mean nocturnal CPAP level averaged epoch by epoch (r = 0.63, p < 0.02). Eighty-eight percent of rapid CPAP augmentations (increases by at least 2 cm H(2)O in less-than-or-equal 2 min) were observed during sleep-wake transitions or after arousals/awakenings (Ar/Aw); 63% of such Ar/Aw were not preceded by any detectable respiratory abnormality. CONCLUSIONS: CPAP levels and variations during auto-CPAP application may be mainly related to sleep continuity and efficiency. The recording of a highly variable pressure during auto-CPAP administration in an unattended environment must raise the question whether the patient's sleep quality was acceptable. A poor sleep quality during an autotitration night could lead to an undesirable overestimation of the CPAP level needed for use with fixed-level CPAP machines. PMID- 11888958 TI - Pulmonary blastomycosis: an appraisal of diagnostic techniques. AB - OBJECTIVES: Pulmonary blastomycosis often mimics bacterial pneumonia or bronchogenic carcinoma, which may result in delayed therapy or the performance of unnecessary diagnostic procedures. We have reviewed the utilization of diagnostic techniques in the workup of patients with pulmonary blastomycosis, defined their diagnostic yields, and proposed an optimal diagnostic approach for the patient in whom pulmonary blastomycosis is considered. DESIGN: Retrospective chart review of all patients with the diagnosis of blastomycosis at a major academic medical center. RESULTS: Of the 119 patients with blastomycosis, 56 (47%) had pulmonary involvement. A total of 92 specimens were obtained by noninvasive means (sputa, 72 specimens; tracheal secretions, 5 specimens; and gastric washings, 15 specimens) in 35 patients. KOH smears were prepared from 22 of those specimens (24%). The diagnostic yield from these culture specimens obtained by noninvasive means was 86% per patient, and 75% per single sample. The diagnostic yields from KOH smears were 46% and 36%, respectively. Flexible bronchoscopy was performed in 24 patients and yielded a diagnosis in 22 (92%). Cultures of bronchial secretions (19 patients) and BAL fluid (6 patients) were positive in 100% and 67% of patients, respectively. The corresponding yields of KOH preparations were 17% (1 of 6 preparations) and 50% (3 of 6 preparations), respectively. Pathology specimens including those from bronchoscopic lung biopsies (nine patients), bronchial brushings (two patients), and bronchoscopic needle aspiration (one patient) were positive in 22%, 50%, and 0% of cases, respectively. Cytology was usually performed to exclude malignancy and was positive for Blastomyces dermatitidis in five patients (sputum, three patients; bronchial washings, two patients). Thoracotomy was performed in 11 cases, and in all patients the procedure yielded a diagnosis. Serology results were available in 25 patients. Immunodiffusion was positive in 10 patients (40%), and complement fixation in 4 patients (16%). CONCLUSIONS: In patients with pulmonary blastomycosis, the positive yield from respiratory specimen cultures is high, but the confirmation of a diagnosis may take up to 5 weeks. Wet smears and cytology examinations of respiratory specimens provide quicker diagnoses but are underutilized. Their routine use is recommended in endemic areas. Commonly used serologic assays are insensitive and are not useful for diagnostic screening. PMID- 11888959 TI - Adequately washed bronchoscope does not induce false-positive amplification tests on bronchial aspirates in the diagnosis of pulmonary tuberculosis. AB - STUDY OBJECTIVES: To investigate the clinical usefulness of amplification (COBAS AMPLICOR; Roche Diagnostics Systems; Branchburg, NJ) on bronchoscopic aspirate specimens in the diagnosis of pulmonary tuberculosis, with particular regard to the possibility of false-positive results in subsequent specimens due to residual Mycobacterium tuberculosis DNA. DESIGN AND SETTING: A prospective clinical study at a tertiary referral medical center. PARTICIPANTS AND METHODS: Four hundred fiberoptic bronchoscopic procedures were performed, using seven bronchoscopes on 335 consecutive patients, for therapeutic or diagnostic purposes. Serial bronchial aspirates were collected and tested for M tuberculosis, using COBAS AMPLICOR (CA). Bronchoscopes were cleaned and disinfected automatically, between patient use, by the same endoscope washer. The name of each bronchoscope and the sequence of its use were recorded, together with the sequence of washing. The CA results were compared with the bacteriologic and histologic results for M tuberculosis infection. When there was a suspicion of contamination, outward polymerase chain reaction analysis was performed. RESULTS: Of 392 specimens (332 subjects), excluding the 8 specimens (4 subjects) in which bacteriologic and histologic analyses were omitted, a smear-positive result for acid-fast bacilli (AFB), culture-positive or biopsy-positive results, and CA-positive results were obtained in 16, 49, and 32 specimens, respectively. In AFB smear-positive subjects, the sensitivity, specificity, positive predictive values (PPVs), and negative predictive values (NPVs) were 92%, 67%, 92%, and 67%, respectively. In AFB smear-negative subjects, the sensitivity, specificity, PPV, and NPV values were 38%, 99%, 74%, and 94%, respectively. The CA test was more sensitive than the AFB smears for the diagnosis of pulmonary tuberculosis (53% vs 27%, respectively; p < 0.05). False-positive CA results were seen in only six specimens. Three of these six subjects received a diagnosis of pulmonary tuberculosis on clinical and radiologic grounds, and none of the six results seemed to be associated with bronchoscopic cross-contamination. CONCLUSIONS: Adequately cleaned and disinfected bronchoscopes did not cause false-positive amplification test results for M tuberculosis on bronchial aspirates by cross contamination. Furthermore, sensitivity was greater with the CA tests. Therefore, CA tests on bronchial aspirates seem to be useful in the diagnosis of pulmonary tuberculosis. PMID- 11888960 TI - Serologic IgE immune responses against Aspergillus fumigatus and Candida albicans in patients with cystic fibrosis. AB - OBJECTIVES: The objectives of this study were to determine the prevalence of Aspergillus fumigatus and Candida albicans in the sputa of patients with cystic fibrosis (CF), to assess serologic IgE responses of these patients to the presence of fungi in the sputum, to evaluate what effect this may have on clinical status, and to determine how the above-mentioned factors relate to allergic bronchopulmonary aspergillosis (ABPA). PATIENTS: Seventy-six CF patients (40 male and 36 female patients; age, 15.3 plus minus 8.7 years [mean plus minus SD]) were studied. MEASUREMENTS AND RESULTS: A total of 1,239 sputum samples from 66 patients were cultured for fungi. A fumigatus was grown in 256 sputum specimens (20.7%), and C albicans was grown in 588 sputum samples (47.5%). Forty patients (60.6%) had at least one positive culture finding for A fumigatus, and 58 patients (87.9%) had at least one positive culture finding for C albicans. Forty-nine patients (64.5%) were sensitized to A fumigatus, and 20 patients (26.7%) were sensitized to C albicans. No correlation was found between the finding of A fumigatus in sputum and IgE to A fumigatus. Only patients who had at least one positive culture finding for C albicans had IgE to C albicans develop. Lung function values and chest radiograph scores were not significantly lower in patients sensitized to either A fumigatus or C albicans as compared to nonsensitized patients. Of the 20 patients sensitized to C albicans, 10 patients had confirmed ABPA and 10 patients had some immunologic characteristics of ABPA. CONCLUSIONS: A high prevalence of colonization and sensitization to A fumigatus and C albicans in CF patients was observed. The sensitization to these fungi was not related to the clinical severity. IgE to C albicans may be an immunologic marker related to the development of ABPA in patients with CF. PMID- 11888961 TI - Bronchial artery embolization : experience with 54 patients. AB - OBJECTIVES: To report our experience with bronchial arteriography and bronchial artery embolization (BAE). MATERIALS AND METHODS: A review of clinical experience to evaluate the demographics, clinical presentation, radiographic studies, bronchoscopy, and complications of bronchial arteriography and BAE at Mayo Medical Center, Rochester, MN, from 1981 to 2000. RESULTS: Fifty-four patients underwent bronchial arteriography. There were 34 men and 20 women with a mean age of 53 years. Hemoptysis was the most common indication in 53 patients (98%). Hemoptysis was caused by bronchiectasis (9 patients), pulmonary hypertension (9 patients), malignancy (7 patients), mycetoma (7 patients), and other identified causes (14 patients). The cause could not be identified in eight patients. Bronchoscopy was performed in 49 patients (92%), and the results identified the bleeding lobe in 32 patients, lateralized the side of the bleeding in 5 patients, and were not helpful in 12 patients. Bronchial arteriography revealed hypervascularity (45 patients), bronchial artery hypertrophy (17 patients), hypervascularity with shunting (15 patients), dense soft tissue staining (8 patients), vascular abnormalities (7 patients), and extravasation of contrast (1 patient). BAE was attempted in 54 patients, completed in 51 patients, and was unsuccessful in 3 patients. Overall, 72 embolization sessions were performed with a total of 131 arteries embolized, and the average number of arteries embolized per patient was 2.5. Control of hemoptysis was observed in 46 patients (85%) at 1 month. Rebleeding occurred within 30 days in five patients. Eight patients had recurrent hemoptysis that occurred 30 days after the procedure. The complications of embolization included subintimal dissection of a bronchial artery (two patients), bronchial arterial perforation by a guidewire (one patient), and the reflux of embolic material into the aorta without adverse sequelae (one patient). CONCLUSIONS: BAE is a useful therapy to control both acute and chronic hemoptysis. BAE may help to avoid surgery in patients who are not good surgical candidates. Should hemoptysis recur in these patients, repeat embolization can be performed safely. PMID- 11888962 TI - Bronchial artery embolization for the management of nonmassive hemoptysis in cystic fibrosis. AB - STUDY OBJECTIVES: Hemoptysis is a common complication in patients with cystic fibrosis (CF). Current approaches to patients with hemoptysis include conservative medical therapy, bronchial artery embolization (BAE), and surgery. We investigated the effectiveness of early BAE on the outcome in patients with minor bleeding. DESIGN, PATIENTS, AND INTERVENTIONS: We reviewed the clinical records from the Cystic Fibrosis Service for eight consecutive patients treated with medical therapy who had undergone early BAE and eight matched patients treated with conservative medical therapy alone. MEASUREMENTS: We assessed the mean number of bleeding episodes, pulmonary exacerbations, lung function (FEV(1)), Shwachman score, and Nottingham Health Profile (NHP) scores, the year before BAE and for the 3 ensuing years. RESULTS: During a 3-year follow-up, patients who underwent embolization had significantly fewer bleeding episodes (p < 0.001) and pulmonary exacerbations (p < 0.006). Lung function declined significantly in both groups (p < 0.001). The modified Shwachman score declined significantly during the follow-up only in patients who did not undergo embolization (p < 0.001). Patients treated by early embolization had significantly better NHP scores, indicating a better quality of life (p < 0.05). None of the patients who underwent BAE had adverse reactions. CONCLUSIONS: Early BAE in patients with CF who have nonmassive hemoptysis is an effective, safe therapeutic approach offering better long-term control of recurrent bleeding and quality of life than medical therapy alone. PMID- 11888963 TI - Estimated incidence of acute pulmonary embolism in a community/teaching general hospital. AB - PURPOSE: This study attempts to determine the incidence of established acute pulmonary embolism (PE) in a community/teaching general hospital. BACKGROUND: The reported incidence of objectively diagnosed acute PE among hospitalized adults in a large urban hospital or major university hospital ranges from 0.27 to 0.40%. Whether the incidence of PE in other categories of hospitals fits within this narrow range is unknown. METHODS: Patients with acute PE diagnosed by ventilation/perfusion lung scan, pulmonary angiography, compression ultrasound in a patient with suspected PE, autopsy, or (by coincidence) lung biopsy were identified among patients hospitalized during a 2-year period from 1998 to 2000. The incidence of PE was also determined according to age, sex, and race. RESULTS: Among adult patients (> or = 20 years old), the incidence of established acute PE was 95 of 34,567 patients (0.27%; 95% confidence interval [CI], 0.22 to 0.34%). No PE was diagnosed in patients < 20 years old. The incidence of PE in men was 36 of 13,722 patients (0.26%; 95% CI, 0.18 to 0.36%); in women, it was 59 of 20,845 patients (0.2%; 95% CI, 0.22 to 0.36%; not significant [NS]). The incidence in African-Americans adults was 10 of 4,344 patients (0.23%; 95% CI, 0.11 to 0.42%); in white adults, it was 84 of 28,615 patients (0.29%; 95% CI, 0.23 to 0.36%; NS). CONCLUSION: The incidence of PE in a community/teaching general hospital was comparable to the incidence in a large urban-care center and in a major university hospital. PMID- 11888964 TI - Treatment of right heart thromboemboli. AB - BACKGROUND: The presence of right heart thromboemboli complicating pulmonary thromboemboli carries with it an increased mortality rate compared to pulmonary thromboemboli alone, but little is known about the optimal management of this difficult clinical situation. This fact is highlighted in the case study of a patient with a 19-cm right atrial thrombus complicating bilateral pulmonary thromboemboli. STUDY OBJECTIVES: We sought to determine the effects of anticoagulation therapy, thrombolysis, and surgical embolectomy on mortality rate in patients with right heart thromboemboli. DESIGN: Retrospective analysis of all reported cases in the English language literature (1966 to 2000) of right heart thromboembolism in which age, sex, therapy, and outcome were reported. MEASUREMENTS AND RESULTS: We analyzed 177 cases of right heart thromboembolism. Pulmonary thromboembolism was present in 98% of the cases. The patients were evenly divided by gender with an average age of 59.8 years (SD, 16.6 years) years. Dyspnea (54.2%), chest pain (22.6%), and syncope (17.5%) were the most common presenting symptoms. The treatments administered were none (9%), anticoagulation therapy (35.0%), surgical procedure (35.6%), or thrombolytic therapy (19.8%). The overall mortality rate was 27.1%. The mortality rate associated with no therapy, anticoagulation therapy, surgical embolectomy, and thrombolysis was 100.0%, 28.6%, 23.8%, and 11.3%, respectively. Using multivariate modeling with survival as the primary outcome, age and gender were not associated with mortality rate, but thrombolytic therapy was associated with an improved survival rate (p < 0.05) when compared either to anticoagulation therapy or surgery. CONCLUSION: The presence of right heart thromboemboli may have diagnostic and therapeutic implications in pulmonary thromboembolism patients. A well-designed prospective, randomized trial is needed to determine the optimal treatment of right heart thromboemboli. PMID- 11888965 TI - Diagnostic value of interleukin-1alpha, interleukin-6, and tumor necrosis factor in pleural effusions. AB - STUDY OBJECTIVES: Interleukin (IL)-1alpha, IL-6, and tumor necrosis factor (TNF) alpha were measured in pleural fluid from 57 patients with pleural effusion in order to evaluate the diagnostic utility of these cytokines. We studied 20 patients with malignant pleural effusion, 11 patients with parapneumonic pleural effusion, 9 patients with tuberculous pleural effusion, and 17 patients with transudative pleural effusion. Cytokines were measured by radioimmunoassay. SETTING: University tertiary hospital. RESULTS: The mean values of the three cytokines measured in pleural fluid or in serum were significantly higher in patients with exudates than with transudates (p < 0.05). The ratio of IL-6 in pleural fluid to serum was significantly higher in exudates than in transudates (p < 0.05). The level of IL-6 in pleural fluid was significantly higher in tuberculous than malignant (p < 0.007) or parapneumonic pleural effusions (p < 0.04). No significant difference between the three types of exudates was found in pleural fluid levels of IL-1alpha or TNF-alpha. CONCLUSIONS: Serum levels of IL 1alpha, TNF-alpha, and in particular IL-6 can distinguish exudates from transudates, while pleural fluid IL-6 levels could be useful as an additional marker in the differential diagnosis of tuberculous, malignant, and parapneumonic exudates. Finally, our results suggest that there is local cytokine production in exudative pleural effusions. PMID- 11888966 TI - Phase II study of repeated intrapleural chemotherapy using implantable access system for management of malignant pleural effusion. AB - STUDY OBJECTIVE: To examine the effectiveness of repeated intrapleural chemotherapy using an implantable access system (INFUSE-A-PORT; Horizon Medical Products; Manchester, GA) for the management of a malignant pleural effusion (MPE). METHODS: Twenty-two patients with histologically proven MPEs (11 men and 11 women; mean age, 63.8 years; age range, 51 to 81 years; performance status, less-than-or-equal 3) were enrolled in this study. There were 17 patients with MPEs resulting from primary lung cancer; of whom 7 had metastatic disease (stage IV), 3 had stage IIIB disease, and 7 had postoperative recurrences. Three patients had MPEs that were caused by mesothelioma, and two had MPEs caused by breast cancer. Intrapleural catheters were placed by a video-assisted thoracoscopic procedure. The intrapleural administration of 5-fluorouracil (5-FU; 250 mg per body) and cisplatin (10 mg per body) using the implantable access system was performed biweekly at the outpatient clinic. Drainage of the MPEs through the port was performed only when the patients had some manifestations that occurred due to increasing effusion. RESULTS: The mean total administered doses of 5-FU and cisplatin were 3,290 and 124 mg, respectively. The mean follow up period was 431 days (range, 209 to 792 days). The median survival period was 403 days, and the longest survival period was 792 days. No treatment-related mortality, renal dysfunction, bone marrow suppression, or infection occurred. One patient experienced a hemothorax after eight instances of intrapleural administration. The port and the catheter were involved with the tumor in one patient. CONCLUSIONS: Repeated intrapleural chemotherapy using the implantable access system is useful and safe for patients with MPEs. In the future, prospective randomized studies will be necessary to compare the efficacy of therapy for MPE using the implantable access system with that of pleurodesis. PMID- 11888967 TI - Clinical significance of pleural effusion in acute aortic dissection. AB - STUDY OBJECTIVE: To clarify the clinical significance of pleural effusion in the clinical course of acute aortic dissection (AAD). DESIGN: Retrospective clinical series. SETTING: A university hospital. PATIENTS: Fifty-five patients strongly suspected of having AAD because of severe chest or back pain. Patients with obvious ischemic heart disease, lung disease, and pleural disease were excluded. INTERVENTIONS: An additional diagnosis of pleural effusion was made when evident by CT. MEASUREMENTS AND RESULTS: Pleural effusion was detected in 42 of 48 patients (88%) with AAD (mean plus minus SD age, 65 plus minus 12 years; 35 men and 13 women), but in only 1 of 7 patients (14%) who proved not to have AAD (mean age, 74 +/- 10 years; 6 men and 1 woman). Effusion appeared at a mean of 4.5 days after onset of dissection. Thoracentesis performed in six patients showed a bloody effusion in three patients and an exudative effusion in three patients. In the six AAD patients without pleural effusion, four patients underwent surgery within 3 days after onset of dissection. In patients with AAD, effusion was demonstrated on the first CT after hospital admission in 18 patients, and was bilateral in 32 patients. WBC count in blood, serum C-reactive protein concentration, and body temperature were higher in patients with effusion (13,400 +/- 3,600/microL, 18.4 +/- 11.5 mg/dL, and 38.2 +/- 0.7 degrees C) than in those without effusion (10,300 +/- 2,900/microL, 4.5 +/- 4.2 mg/dL, and 37.0 +/- 1.0 degrees C, respectively). CONCLUSIONS: Pleural effusion occurs frequently in patients with AAD, often in association with inflammatory reactions. PMID- 11888968 TI - Suction vs water seal after pulmonary resection: a randomized prospective study. AB - STUDY OBJECTIVE: To evaluate whether suction or water seal is superior in the management of chest tubes after pulmonary resection. DESIGN: A prospective, randomized, controlled trial. After an initial, brief period of suction, patients were randomized to water seal or - 20 cm H(2)O suction. SETTING: University hospital. PATIENTS: Sixty-eight patients who underwent wedge resection, segmentectomy, or lobectomy were included in the study. Those patients who underwent reoperative surgery or lung volume reduction surgery were excluded. RESULTS: There were 34 patients in each group. The two groups were evenly matched for age, sex, operation performed, severity of lung disease, and nutritional status. Fifteen patients in each group (44%) had an air leak at the completion of surgery. The duration of the air leak was shorter in the water seal group than in the suction group (mean +/- SEM, 1.50 +/- 0.32 days vs 3.27 +/- 0.80 days, respectively; p = 0.05). The mean times to removal of chest tubes were 3.33 +/- 0.35 days in the water seal group and 5.47 +/- 0.98 days in the suction group (p = 0.06). The length of stapled parenchyma was measured for each patient and averaged 24.9 cm for the water seal group and 18.5 cm for the suction group (p = 0.18). When corrected for the length of staple lines, the duration of air leaks and days with chest tube were dramatically lower in the water seal group (p = 0.02 and p = 0.02, respectively). CONCLUSION: Placing chest tubes on water seal after a brief period of suction after pulmonary resection shortens the duration of the air leak and likely decreases the time that the chest tubes remain in place. Adoption of this practice may result in lower morbidity and lower hospital costs. PMID- 11888969 TI - Short-term course and outcome of treatments of pleural empyema in pediatric patients: repeated ultrasound-guided needle thoracocentesis vs chest tube drainage. AB - BACKGROUND: Several reports have suggested that early chest tube drainage (CTD) may not be necessary in the treatment of severe pleural empyema (PE) in pediatric patients if appropriate antibiotic therapy and supportive care are provided. OBJECTIVES: A prospective open study to compare the short-term course of two treatment protocols of severe PE in pediatric patients. STUDY DESIGN: One group of 32 patients was treated with early insertion of a chest tube for CTD, and a second group of 35 patients was treated by a repeated ultrasound-guided needle thoracocentesis (RUSGT). The severity of the empyema was assessed by chest radiograph, the amount of fluid drained, the number of days the patient had experienced a fever, and the duration of antibiotic treatment. RESULTS: No significant differences were found between the two groups (RUSGT vs CTD) in all of the following measurements: mean (plus minus SD) duration of a temperature > or = 39 degreesC, 6.2 +/- 2.4 vs 6.5 +/- 1.8 days, respectively; mean duration of a temperature > or = 38 degreesC, 9 +/- 3.9 vs 8.2 +/- 4.5 days, respectively; fluid drained, 35.1 + 23.8 vs 30 +/- 28.2 mL/kg, respectively; duration of antibiotic treatment, 30 +/- 13.2 vs 30.2 +/- 7.3 days, respectively; and length of hospitalization and home IV treatment, 22 +/- 7.6 vs 24.2 +/- 7.5 days, respectively. A failure to respond to treatment occurred in three patients in the RUSGT-treated group and in five patients in the CTD-treated group. The failure to respond occurred in the RUSGT-treated group only in those patients with very large empyemas that caused mediastinal deviation. CONCLUSION: The treatment of PE by RUSGT is as efficacious as CTD, unless PE causes mediastinal deviation. PMID- 11888970 TI - The relationship between left ventricular function assessed by multigated radionuclide test and cardiopulmonary exercise test in patients with ischemic heart disease. AB - STUDY OBJECTIVES: To compare the oxygen pulse curve (O(2)P-C) as measured during cardiopulmonary exercise testing (CPET) with left ventricular (LV) ejection fraction (LVEF) rest-exercise response as measured by multigated equilibrium (99m)Tc radionuclide cineangiography (MUGA) in patients with different degrees of ischemic heart disease (IHD). PATIENTS: Forty-six patients (39 men and 7 women; mean plus minus 1 SD age, 59.2 plus minus 11 years) with IHD, with no hypertrophic, valvular, or pericardial disease. METHODS: A supine bicycle ergometer with increments of 25 W every 2 min was used for MUGA, and an electronically braked cycle ergometer was used for upright symptoms-limited CPET. Exercise was increased by 10 to 20 W/min until the target heart rate (HR) was reached (similar peak HR for both studies). MEASUREMENTS AND RESULTS: The O(2)P-C was scored on a 10-point scale as follows: type A, normal curve (10 points); type B, normal-shaped curve with low values (8 points); type C, low and flat curve (5 points); type D, descending curve (3 points). Findings for the MUGA study were classified into four groups by the degree of ischemic response: group 1 (control), normal diastolic function (n = 10), LVEF > 55%, LVEF during exercise minus LVEF at rest [DeltaLVEF] greater-than-or-equal 5%; group 2, mild ischemia (n = 10), LVEF > 55%, < 0 DeltaLVEF < 5%, diastolic dysfunction at exercise (prominent "A" waves); group 3, LV dysfunction (n = 9), LVEF < or = 35% at rest; and group 4, significant ischemia (n = 17), LVEF > 55%, DeltaLVEF < 0, diastolic dysfunction. A highly significant relationship between the O(2)P-C score and the MUGA grouping was observed by Fisher's Exact Test and Pearson's linear regression line (p < 0.001; R = - 0.89). CONCLUSIONS: Exercise-responded O(2)P-C might serve as a good noninvasive, physiologically based, parameter to distinguish between IHD patients with normal and impaired LV function. PMID- 11888971 TI - Association between inhaled beta-agonists and the risk of unstable angina and myocardial infarction. AB - BACKGROUND: beta-Adrenoceptor agonists (beta-agonists) are commonly used to treat obstructive lung diseases, and preliminary studies have suggested they are associated with an increased risk of adverse cardiovascular outcomes. We further examined the association between acute coronary syndromes and inhaled beta agonist therapy. METHODS: We performed a nested, case-control study using data that were collected as part of a larger, ongoing, prospective study of quality improvement in the primary care clinics of seven Veterans Administration Medical Centers. We identified 630 patients with unstable angina or acute myocardial infarction hospitalized between 1996 and 1999. We frequency matched these case patients to 10,486 control subjects according to clinic location, and randomly assigned each an "index date." The computerized pharmacy database at each center was used to ascertain beta-agonist use. Cardiovascular risk factors were assessed from mailed questionnaires and electronic medical records, which included inpatient and outpatient diagnoses, medications, and laboratory results. RESULTS: In comparison with patients who had not filled a beta-agonist prescription during the 90 days prior to their index date, patients who had filled a beta-agonist prescription had an increased risk of experiencing an acute coronary syndrome. The increased risk of an acute coronary syndrome persisted after adjusting for age and cardiovascular risk factors, including hypertension, diabetes, and smoking history. Moreover, there was a dose-response relationship with an adjusted odds ratio (OR) of 1.38 for one to two metered-dose inhaler (MDI) canisters (95% confidence interval [CI], 0.86 to 2.23), an OR of 1.57 for three to five MDI canisters (95% CI, 1.01 to 2.46), and an OR of 1.93 for six or more MDI canisters (95% CI, 1.23 to 3.03). After stratifying for receipt of a beta blocker prescription, the adjusted OR in subjects who did not receive a beta blocker was 1.55 for one to two MDI canisters (95% CI, 0.60 to 3.99), an OR of 4.07 for three to five canisters (95% CI, 2.17 to 7.64), and an OR of 3.83 for six or more canisters (95% CI, 2.02 to 7.29). Subjects who had received both beta blockers and beta-agonists had no increase in risk in acute coronary syndromes unless they had filled six or more beta-agonist MDI canisters. CONCLUSIONS: A prescription for inhaled beta-agonists may increase the risk of myocardial infarction and unstable angina in patients with COPD. PMID- 11888972 TI - Parathyroid hormone-related protein in epithelial lining fluid in humans negatively correlates with the severity of lung injury. AB - OBJECTIVE: To determine the concentration of parathyroid hormone-related protein (PTHrP; an autocrine/paracrine regulator of type-2 alveolar epithelial cells proliferation and apoptosis) in the epithelial lining fluid (ELF) from patients without pulmonary disease and from patients with acute lung injury (ALI), and to evaluate whether PTHrP concentrations correlated with the intensity of lung injury. DESIGN: Prospective study. SETTING: An adult trauma/surgical ICU in an urban teaching hospital. PATIENTS: A total of 20 patients with ALI receiving mechanical ventilation (patients), and 10 patients without pulmonary disease not receiving mechanical ventilation (control subjects). INTERVENTIONS: None. MEASUREMENTS AND RESULTS: PTHrP was detected in all BAL fluids, and ELF PTHrP concentrations (median; 25% to 75% percentiles) tended to be higher in patients (52.2 nmol/mL; 20.8 to 65.6 nmol/mL) than in control subjects (25.4 nmol/mL; 20.5 to 35.4 nmol/mL; p = 0.18). In patients, ELF PTHrP concentration correlated positively with the PaO(2)/fraction of inspired oxygen ratio (r = 0.53; p = 0.005), and negatively with lung injury score (r = - 0.44; p = 0.02), radiologic score (r = - 0.40; p = 0.04), and BAL albumin concentration (r = - 0.42; p = 0.02). CONCLUSION: PTHrP is present in biologically significant concentrations in the alveolar milieu in humans. In patients with ALI, the PTHrP concentration correlates negatively with the degree of lung injury. PMID- 11888973 TI - A randomized clinical trial of intermittent subglottic secretion drainage in patients receiving mechanical ventilation. AB - OBJECTIVE: To study the effect of subglottic secretions drainage on the incidence of ventilator-associated pneumonia (VAP) in patients receiving mechanical ventilation. DESIGN: A randomized clinical trial. SETTING: A 12-bed general ICU. PATIENTS: One hundred fifty patients with an expected duration of mechanical ventilation > 72 h were enrolled in the study. INTERVENTION: Patients were randomly assigned to receive either an endotracheal tube for intermittent subglottic secretions drainage or a standard endotracheal tube. OUTCOME MEASUREMENTS: Incidence of VAP, duration of mechanical ventilation, length of ICU stay, length of hospital stay, and mortality. RESULTS: Seventy-five patients were randomized to subglottic secretion drainage, and 75 patients were randomized to the control group. The two groups were similar at the time of randomization with respect to demographic characteristics and severity of illness. VAP was seen in 3 patients (4%) receiving suction secretion drainage and in 12 patients (16%) in the control group (relative risk, 0.22; 95% confidence interval, 0.06 to 0.81; p = 0.014). The other outcome measures were not significantly different between the two groups. CONCLUSION: Intermittent subglottic secretion drainage reduces the incidence of VAP in patients receiving mechanical ventilation. PMID- 11888975 TI - Accuracy of three electronic monitors for metered-dose inhalers. AB - BACKGROUND: Prior studies indicate that some devices used to monitor metered-dose inhaler (MDI) use are not accurate. OBJECTIVE: To assess the accuracy of the Doser CT (NEW-MED Corporation; Waltham, MA), the MDILog (Medtrac Technologies; Lakewood, CO), and the SmartMist (Aradigm Corporation; Hayward, CA) in a bench top study. DESIGN: One, two, and four puffs of fluticasone propionate MDI (Flovent; GlaxoWellcome; Research Triangle Park, NC) were actuated twice daily for 30 days with two units of each device. The date and time of each actuation were recorded in a log and then compared with the output of the device. The percentage of doses recorded accurately was compared by analysis of variance. MEASUREMENTS AND RESULTS: The SmartMist was 100% accurate, while both the Doser CT and MDILog devices occasionally recorded spurious actuations. The Doser CT also had missed actuations after the counter had prematurely reached zero secondary to the spurious recordings. The accuracy (mean plus minus SD) was 94.3 plus minus 2.9% for the Doser CT and 90.1 plus minus 6.9% for the MDILog (p = 0.21). The dose regimen actuated and duration of use did not significantly affect accuracy. CONCLUSION: All three devices are sufficiently accurate to monitor adherence in most clinical settings. PMID- 11888974 TI - Silver-coated endotracheal tubes associated with reduced bacterial burden in the lungs of mechanically ventilated dogs. AB - STUDY OBJECTIVES: To evaluate the influence of silver-coated endotracheal tubes on the lung bacterial burden of mechanically ventilated dogs. DESIGN: Randomized, double-blinded, controlled experiment. SETTING: Animal research facility of a regional medical university. PATIENTS OR PARTICIPANTS: Eleven healthy adult dogs. INTERVENTIONS: The dogs were intubated either with cuffed, noncoated endotracheal tubes or with endotracheal tubes having a novel antimicrobial silver hydrogel coating and were challenged with buccal administration of Pseudomonas aeruginosa. MEASUREMENTS AND RESULTS: The silver coating delayed the appearance of bacteria on the inner surface of the endotracheal tubes ([mean +/- SD] duration of mechanical ventilation before appearance of bacteria, 3.2 +/- 0.8 days; mean duration of mechanical ventilation, 1.8 +/- 0.4 days; p = 0.016). The mean total aerobic bacterial burden in the lung parenchyma was statistically lower among the dogs receiving the silver-coated endotracheal tubes compared to those not receiving them (4.8 +/- 0.8 vs 5.4 +/- 9 log cfu/g lung tissue, respectively; p = 0.010). Pronounced differences were seen in the gross and histologic assessments of inflammation in the lung. Using an increasing severity scale of 0 to 12 to assess four components of histology (ie, hyperemia, edema, cellular infiltration, and bacterial presence), dogs receiving noncoated endotracheal tubes had statistically greater histology scores compared to dogs receiving silver-coated endotracheal tubes (7.1 plus minus 1.6 vs 2.8 plus minus 1.2, respectively; p < 0.001). CONCLUSION: These results suggest that the silver coating of endotracheal tubes may delay the onset of and decrease the severity of lung colonization by aerobic bacteria. Based on these results, clinical studies are planned to determine the safety and clinical efficacy of silver-coated endotracheal tubes in patients requiring mechanical ventilation in the ICU setting. PMID- 11888976 TI - Major pulmonary embolism: review of a pathophysiologic approach to the golden hour of hemodynamically significant pulmonary embolism. AB - Major pulmonary embolism (PE) results whenever the combination of embolism size and underlying cardiopulmonary status interact to produce hemodynamic instability. Physical findings and standard data crudely estimate the severity of the embolic event in patients without prior cardiopulmonary disease (CPD) but are unreliable indicators in patients with prior CPD. In either case, the presence of shock defines a threefold to sevenfold increase in mortality, with a majority of deaths occurring within 1 h of presentation. A rapid integration of historical information and physical findings with readily available laboratory data and a structured physiologic approach to diagnosis and resuscitation are necessary for optimal therapeutics in this "golden hour." Echocardiography is ideal because it is transportable, and is capable of differentiating shock states and recognizing the characteristic features of PE. Spiral CT scanning is evolving to replace angiography as a confirmatory study in this population. Thrombolytic therapy is acknowledged as the treatment of choice, with embolectomy reserved for those in whom thrombolysis is contraindicated. PMID- 11888977 TI - Interaction of hemostatic genetics with hormone therapy: new insights to explain arterial thrombosis in postmenopausal women. AB - Genetic variants of key hemostatic mediators increasingly have been proposed as risk factors for atherothrombosis. The Hormone and Estrogen/Progestin Replacement Study group recently reported that the initiation of estrogen replacement in postmenopausal women with known coronary heart disease is associated with an early increase in cardiovascular events. A putative genetic susceptibility factor has been proposed a potential mediator of this increased event risk. This review outlines the recent literature to support the premise for this important proposal. Genetic profiling has great potential to improve the safety and efficacy of individualized pharmacotherapy in postmenopausal women and other at risk populations for the prevention of cardiovascular disease. PMID- 11888978 TI - Corticosteroids and cardiopulmonary bypass : a review of clinical investigations. AB - Traditionally, corticosteroids have been administered to patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) to ward off detrimental physiologic alterations associated with activation of the systemic inflammatory response, yet few well-controlled investigations exist, and use of these drugs in this setting remains controversial. This review article critically examines the results of clinical investigations in this area, and certain conclusions are suggested. The constellation of findings indicate that corticosteroids offer no clinical benefits to patients undergoing cardiac surgery with CPB and in fact may be detrimental. Further directions for clinical research in this area are also suggested. PMID- 11888979 TI - Treating tobacco use and dependence: an evidence-based clinical practice guideline for tobacco cessation. AB - The prevention of tobacco-related morbidity and mortality through smoking cessation intervention is among the most vital missions of the chest clinician. This article summarizes the major findings and clinical recommendations of the US Department of Health and Human Services/Public Health Service Guideline, Treating Tobacco Use and Dependence, which is a comprehensive, evidence-based blueprint for smoking cessation. By becoming fluent in the clinical interventions and by implementing the simple institutional changes described in this article and in the guideline, chest clinicians can more effectively intervene with their patients who smoke. PMID- 11888980 TI - Using the chest radiograph to determine intravascular volume status: the role of vascular pedicle width. AB - Due to concerns about the efficacy and safety of using pulmonary artery catheterization to evaluate hemodynamic status, noninvasive diagnostic testing has gained increased importance. This article focuses on both the supportive evidence and the limitations of applying the vascular pedicle width (VPW), which is the mediastinal silhouette of the great vessels, as an aid in the assessment of patients' intravascular volume status. The objective measurement of the VPW obtained from either upright or supine chest radiographs (CXRs which are often already available though not fully utilized) can increase the accuracy of the clinical and radiographic assessment of intravascular volume status by 15 to 30%, and this value may be even higher when VPW is used serially within the same patient. Regardless of the presence or absence of pulmonary edema, the best VPW cutoff for differentiating a high vs normal to low intravascular volume status is 70 mm. Patients with a VPW of > 70 mm coupled with a cardiothoracic ratio of > 0.55 are more than three times more likely to have a pulmonary artery occlusion pressure > 18 mm Hg than are patients without these radiographic findings. We suggest a management algorithm for utilizing the VPW, and whether or not such an approach will offer superior patient outcomes requires prospective investigation. Reappraisal of the VPW and other roentgenographic signs should be incorporated into newly implemented studies of the Swan-Ganz catheter, ICU echocardiography, portable CT scans, and other costlier technologies. While such investigations may refine the optimum application of the portable CXR, conventional and digital supine radiographs should retain an important role in the diagnosis and management of critically ill patients. Lastly, the measurement of the VPW should be incorporated into the training of chest clinicians and radiologists. PMID- 11888981 TI - Comparison of the postoperative blood flow waveforms of the bypassing grafts in patients following minimally invasive direct coronary artery bypass. AB - PURPOSE: To use Doppler ultrasound velocimetry to detect and compare the postoperative flow characteristics of the bypassing grafts in patients following minimally invasive direct coronary artery bypass surgery (MIDCAB). MATERIALS AND METHODS: From January 1997 to June 1999, 34 patients underwent MIDCAB with the left internal thoracic artery (LITA) to the left anterior descending coronary artery (LAD) [n = 23], with the right gastroepiploic artery (RGEA) to the right posterior descending artery (RPD) [n = 3], or with the LITA with a saphenous vein graft extension to the LAD (n = 6), the diagonal coronary artery (n = 1), or the right acute coronary artery (n = 1). There were two patients with LITA to the LAD and RGEA to the RPD. Patients underwent MIDCAB due to coronary artery stenosis (100% occlusion, n = 10; 90 to 99% stenosis, n = 18; < 90% stenosis, n = 5) or unsuccessful percutaneous transcoronary angioplasty with dissection (n = 1). All patients underwent flow velocity measurement by Doppler ultrasound velocimetry in the immediate postoperative period, and at 6-month and 12-month intervals; graft flows were quantified based on Doppler velocimetric data. RESULTS: The results showed that in a patient with a totally occluded LAD or RPD, typical biphasic velocity waveforms were consistently observed. However, a delayed diastolic wave was noted in RGEA grafts. In patients with less-occluded stenotic lesions or with strong back flows, the flow velocity patterns showed biphasic waveforms but systolic reversal was observed in the area closest to the anastomotic site. CONCLUSION: The presence of an LAD or RPD stenosis proximal to the anastomotic site significantly affects the LITA or RGEA graft flow volume. The biphasic flow pattern proves that an LITA or RGEA graft transports the blood primarily to coronary arteries during the diastolic phase. PMID- 11888982 TI - Do specialists differ on do-not-resuscitate decisions? AB - STUDY OBJECTIVE: Opinions regarding do-not-resuscitate (DNR) decisions differ between individual physicians. We attempted to determine whether the strength of DNR recommendations varies with medical specialty and experience. DESIGN: Written survey. PARTICIPANTS: Physicians from the pulmonary/critical-care medicine (PCCM), cardiology, internal medicine, gastroenterology, hematology/oncology, and infectious disease services as well as the Department of Medicine house staff at our tertiary-care referral center participated in the study. INTERVENTIONS: Physicians were asked confidentially to quantify the strength of their opinions on discussing and recommending DNR orders for each of 20 vignettes made from the summaries of actual cases. Reasons for their opinions and demographic data also were recorded. MEASUREMENTS AND RESULTS: One hundred fifteen of 155 physicians (74%) responded. PCCM physicians (mean [+/- SD] DNR score, 157 +/- 22) more strongly recommended DNR orders than cardiologists (mean DNR score, 122 +/- 32; p = 0.006), house staff (mean DNR score, 132 +/- 24; p = 0.014), and general internists (mean DNR score, 129 +/- 30; p = 0.043). PCCM physicians also trended toward recommending DNR orders for more of the 20 patients described in the vignettes compared to cardiologists (mean DNR number, 16.5 +/- 3.0 vs 11.9 +/- 5.8, respectively; p = 0.066). There were no differences between PCCM physicians and hematology/oncology, infectious disease, and gastroenterology specialists. Among the house staff, the likelihood of recommending a DNR order correlated significantly with increasing years of experience (r = 0.45; p = 0.002). The opposite trend was present in the specialty staff groups. No significant differences in opinion by gender, religion, or personal experiences were found. CONCLUSIONS: The strength of DNR order recommendations varies with medicine specialty and years of training and experience. An awareness of these differences and the determination of the reasons behind them may help to target educational interventions and to ensure effective collaboration with colleagues and communication with patients. PMID- 11888983 TI - Renovascular hypertension: problems in evaluation and management. PMID- 11888984 TI - Fatal work-related inhalation of harmful substances in the United States. AB - STUDY OBJECTIVES: Inhalation of harmful substances is common in the workplace. The purpose of this study was to describe the epidemiology of fatal occupational inhalations in the United States. DESIGN: Data from the Census of Fatal Occupational Injuries from 1992 to 1998 were analyzed. Information on demographic characteristics, occupation, and industry was used to calculate specific mortality rates, and the inhaled substances were identified. RESULTS: Nationwide, there were 523 cases of fatal occupational inhalation, with a mortality rate of 0.56 deaths per 1,000,000 worker-years. The rate of death was greater for men (1.01/1,000,000) than for women (0.03/1,000,000), and workers > or = 65 years of age had the highest mortality. Mining was the industry with the highest mortality rate (6.64/1,000,000). The occupations with the highest rate were firefighters (3.54/1,000,000) and farming, forestry, and fishing occupations (2.84/1,000,000). Nearly half of the inhalation victims were constructing, repairing, cleaning, inspecting, or painting when the injury occurred. Overall, carbon monoxide was the most frequently inhaled substance (33.5%). The incidence of fatal carbon monoxide inhalations was twice as high in the winter as in the summer. The proportion of workers killed by carbon monoxide poisoning increased with increasing age. CONCLUSIONS: Work-related inhalations cause more deaths than any other mode of exposure to harmful substances. Recognizing those circumstances that pose a higher risk for maintenance and repair workers, as well as upgrading carbon monoxide poisoning prevention programs, could have a major impact in reducing fatal work-related inhalation injuries. PMID- 11888985 TI - An outbreak of bronchoscopy-related Mycobacterium tuberculosis infections due to lack of bronchoscope leak testing. AB - BACKGROUND: Bronchoscopy-related transmission of Mycobacterium tuberculosis is rarely reported. In August 1999, five M tuberculosis-positive bronchial washing culture findings were noted in patients who underwent bronchoscopy in July in a hospital that reported only eight M tuberculosis-positive culture findings from 1995 to 1998, prompting further investigation. METHODS: A case was defined as a M tuberculosis-positive culture finding from specimens obtained from patients who underwent bronchoscopy during January to August of 1999. Bronchoscopy and laboratory records, procedures, and practices were reviewed. M tuberculosis isolates were compared using restriction fragment length polymorphism (RFLP) analysis. RESULTS: During July 1999, 19 bronchoscopic procedures were performed in 19 patients. Bronchial washing specimens for mycobacterial culture were obtained from 18 patients. Ten cases were identified. Two case patients, including the index patient, had signs and symptoms of active tuberculosis prior to bronchoscopy. M tuberculosis infections developed in two more case patients despite starting a standard four-drug antituberculous regimen within 3 weeks after bronchoscopy. Six case patients had positive culture findings but no evidence of infection. All M tuberculosis isolates were antituberculosis-drug susceptible, and all but one were indistinguishable by RFLP analysis. Three bronchoscopes were used during the outbreak period; one bronchoscope was used in 9 of the 10 case patients (relative risk, 8.1; 95% confidence interval, 1.3 to 52). A hole was discovered in the sheath of this bronchoscope. Leak testing, a critical step in bronchoscope reprocessing, was not routinely performed at this institution. CONCLUSIONS: M tuberculosis contamination of the bronchoscope occurred during the index patient's procedure. The hole in the sheath provided access to a space that was difficult to mechanically clean and chemically disinfect. The reprocessing recommendations of bronchoscope manufacturers, including leak testing after each use, should be closely followed. PMID- 11888986 TI - A 62-year-old man with multiple pulmonary nodular opacities. PMID- 11888987 TI - Intraoperative detection of a bronchial carcinoid with a radiolabeled somatostatin analog. AB - Carcinoid tumors of the lung are rare neuroendocrine tumors that make up approximately 1 to 2% of all lung neoplasms. These tumors overexpress somatostatin receptors, and somatostatin analog therapy has become standard in the treatment of carcinoid tumors. In addition, radiolabeled somatostatin analogs have been used to diagnose and treat these lesions. We describe the case of a patient with a right lung mass diagnosed as a carcinoid tumor. The patient underwent complete resection of this tumor with the assistance of intraoperative detection with a handheld gamma probe after the administration of the radiolabeled somatostatin analog (111)In-pentetreotide. This approach allowed us not only to detect the tumor easily, but to scan the bed of the tumor after resection and to re-excise an area of increased radioisotope uptake that corresponded to the presence of residual tumor. We believe this to be the first reported case of bronchial carcinoid resected with the assistance of intraoperative gamma detection after the administration of a radiolabeled somatostatin analog. This technology allowed us to achieve a complete surgical resection with no residual tumor detected either pathologically or by somatostatin scanning. PMID- 11888988 TI - Management of an intrabronchial rupture of right main pulmonary artery: a case report. AB - We report a case of massive endobronchial hemorrhage following a fistula formation of the right pulmonary artery to the right mainstem bronchus in a 15 year-old girl. The fistula had occurred 39 days after the patient had undergone bilateral lung transplantation for cystic fibrosis. The post-transplantation course was remarkable for bronchial colonization by Aspergillus at the site of right bronchial anastomosis and an episode of spontaneous, self-limited hemoptysis on postoperative day 17. A massive endobronchial hemorrhage during surveillance bronchoscopy occurred 39 days after transplantation. Immediate intervention, including rigid bronchoscopy followed by surgery, was effective in saving the patient. The pathophysiologic hypothesis to explain the fistula of the right pulmonary artery to the right mainstem bronchus probably involves ischemia of the anastomosis with necrosis of the suture zone complicated by endobronchial infection with Aspergillus. Rigid bronchoscopic intervention associated with an excellent medical surgical collaboration was pivotal in successfully rescuing the patient. PMID- 11888989 TI - Severe upper airway obstruction from cricoarytenoiditis as the sole presenting manifestation of a systemic lupus erythematosus flare. AB - Upper airway obstruction due to laryngeal involvement is a known complication of systemic lupus erythematosus (SLE). Laryngeal involvement typically accompanies inflammatory activity involving other sites and varies from mild mucosal inflammation to bilateral vocal cord immobility. Cricoarytenoid arthropathy is a rare cause of severe airway obstruction in patients with SLE and almost always occurs in the presence of other associated symptoms. Furthermore, in contrast to patients with rheumatoid arthritis, in whom chronic involvement of cricoarytenoid joints occurs more commonly and often requires surgical intervention, patients with SLE typically present with acute arthritis of cricoarytenoid joints and respond to corticosteroid therapy alone. We describe a patient with known SLE who presented with severe acute upper airway obstruction as the sole manifestation of active SLE after several years of quiescence. The laryngeal involvement progressed from mucosal inflammation to acute cricoarytenoiditis, despite the administration of high-dose corticosteroid therapy, necessitating emergent intubation and tracheostomy. This case illustrates the importance of considering SLE in the differential diagnosis of patients presenting with acute upper airway obstruction. PMID- 11888990 TI - Coronary artery air embolism complicating a CT-guided transthoracic needle biopsy of the lung. AB - Systemic air embolism is a rare but potentially fatal complication of percutaneous transthoracic needle biopsy of the lung. Coronary air embolism can result in myocardial infarction, cardiac arrest, or dysrhythmias. We present the first case of cardiac arrest and myocardial infarction confirmed by ECG and cardiac enzymes in the presence of air in the left coronary artery documented by CT scan in a 77-year-old man after CT-guided transthoracic needle biopsy of the lung. PMID- 11888991 TI - Use of urine pregnancy test for rapid diagnosis of primary pulmonary choriocarcinoma in a man. AB - Primary pulmonary choriocarcinoma is an extremely rare tumor in men, with 13 cases reported in the literature. Due to its rarity, primary choriocarcinoma of the lung in men is often incorrectly diagnosed as more common diseases, such as primary or metastatic lung cancer, and therefore potentially curative chemotherapy or surgery may be withheld from the patient. In this report, we present the case of a 23-year-old man with hemoptysis and progressive dyspnea. Airspace consolidation with multiple nodules of varying sizes was found on a chest radiograph. The results of a urine pregnancy test were positive, and the beta-human chorionic gonadotropin level was markedly elevated both in the serum and the urine. Subsequently, testing of a bronchoscopic biopsy specimen proved these tumors to be choriocarcinoma. We conclude that the urine pregnancy test, a simple and convenient method, would be very useful in the rapid diagnosis of primary pulmonary choriocarcinoma in men. PMID- 11888992 TI - Tracheal compression by the stomach following gastric pull-up: diagnosis with CT and treatment with expandable metallic stent placement. AB - Surgical treatment of recurrent achalasia includes esophagectomy with gastric pull-up. A MEDLINE search yielded no articles describing an adverse effect of this surgery on pulmonary function. We report the first case of acute ventilatory failure caused by gastric pull-up. An evaluation by flexible bronchoscopy, spirometry with flow-volume loops, and dynamic CT scanning revealed extrinsic compression of the trachea by the stomach causing obstruction. Endotracheal placement of a self- expanding stent resulted in the rapid extubation of the patient with normalization of the flow-volume loop and dramatic improvement in the FVC, FEV(1), and peak expiratory flow. PMID- 11888994 TI - MIASMA: asthma exacerbation reduction with salmeterol. PMID- 11888993 TI - What is a spontaneous pneumothorax? PMID- 11888995 TI - Sarcoidosis and gas exchange measures. PMID- 11888996 TI - Diagnosis of pleural tuberculosis. PMID- 11888997 TI - Latent traumatic diaphragmatic hernia: a surgical challenge. PMID- 11888998 TI - Interpretation of home oximetry tracings. PMID- 11888999 TI - Changes in oxygen saturation in patients undergoing fiberoptic bronchoscopy. PMID- 11889000 TI - Revisiting thoracentesis procedures. PMID- 11889005 TI - Modulating cardiac hypertrophy by manipulating myocardial lipid metabolism? PMID- 11889006 TI - Smoking a single cigarette rapidly reduces combined concentrations of nitrate and nitrite and concentrations of antioxidants in plasma. AB - BACKGROUND: Cigarette smoking is a well-known risk factor for the development of cardiovascular disease, yet the mechanism of action involved is not completely understood. Because cigarette smoke contains superoxide and other reactive oxygen species, it has been hypothesized that some of the adverse effects of smoking may result from oxidative damage to endothelial cells, which results in nitric oxide (NO) shortage. However, little information is available regarding the acute effects of smoking on plasma concentrations of NO and antioxidants. We measured the changes in the combined plasma concentrations of nitrate and nitrite as an index of NO concentration, as well as changes in concentrations of major serum antioxidants (ascorbic acid, cysteine, methionine, and uric acid) in smokers after smoking a single cigarette. METHODS AND RESULTS: A randomized crossover study of the effects of smoking a single cigarette was performed in 20 smokers. Smoking a sham cigarette induced no significant changes in all assayed parameters. However, smoking a single cigarette significantly decreased nitrate and nitrite plasma concentrations by 3.5 +/-1.2 and 3.4 +/- 1.1 micromol/L, compared with plasma concentrations at presmoking and sham smoking, respectively. The concentrations of ascorbic acid and other antioxidants were also significantly lower after smoking a single cigarette. These parameters returned to preexperimental levels 60 minutes after smoking cessation. CONCLUSION: The present findings indicate that smoking a single cigarette temporarily decreases nitrate, nitrite, and serum antioxidant concentrations in the plasma. These transient changes may partially contribute to coronary vasoconstriction, which is routinely observed after smoking. PMID- 11889007 TI - Expression of toll-like receptors in human atherosclerotic lesions: a possible pathway for plaque activation. AB - BACKGROUND: Innate immune reactions against bacteria and viruses have been implicated in the pathogenesis of atherosclerosis. To explore the molecular mechanism by which microbe recognition occurs in the artery wall, we characterized the expression of toll-like receptors (TLRs), a family of pathogen pattern recognition receptors, in atherosclerotic lesions. METHODS AND RESULTS: Semiquantitative polymerase chain reaction and immunohistochemical analysis demonstrated that of 9 TLRs, the expression of TLR1, TLR2, and TLR4 was markedly enhanced in human atherosclerotic plaques. A considerable proportion of TLR expressing cells were also activated, as shown by the nuclear translocation of nuclear factor-kappaB. CONCLUSION: Our findings illustrate a repertoire of TLRs associated with inflammatory activation in human atherosclerotic lesions, and they encourage further exploration of innate immunity in the pathogenesis of atherosclerosis. PMID- 11889008 TI - Relationship between lipid levels and clinical outcomes in the Long-term Intervention with Pravastatin in Ischemic Disease (LIPID) Trial: to what extent is the reduction in coronary events with pravastatin explained by on-study lipid levels? AB - BACKGROUND: The Long-Term Intervention with Pravastatin in Ischemic Disease (LIPID) trial showed that pravastatin significantly reduced mortality and coronary heart disease (CHD) events in 9014 patients with known CHD and total cholesterol 4.0 to 7.0 mmol/L at baseline. Secondary objectives included assessment of CHD event reduction according to lipid levels. METHODS AND RESULTS: We investigated the relationships of baseline and on-study lipids with subsequent CHD events in separate Cox models. Treatment effect on CHD event reduction was examined by baseline lipids and after adjustment for on-study lipid levels. Baseline lipids were significant predictors of CHD events. The adjusted relative risk per mmol/L (on placebo) was 1.24 (P=0.004) for total cholesterol, 1.28 (P=0.002) for low-density lipoprotein cholesterol, and 0.52 (P=0.004) for high density lipoprotein cholesterol. Apolipoproteins A1 and B were strong predictors (each P=0.001). Pravastatin reduced the risk of the composite outcome of fatal CHD or nonfatal myocardial infarction by 24% (95% confidence interval [CI], 15% to 32%) and the expanded end point of fatal CHD, nonfatal myocardial infarction, unstable angina, or coronary revascularization by 17% (95% CI, 10% to 24%). Similar relative effects were observed for different categories of baseline lipids. The proportion of treatment effect explained by on-study lipid levels was 67% (95% CI, 27% to 106%) for the composite and 97% (95% CI, 49% to 145%) for the expanded end point. The most important lipids associated with event reduction were apolipoprotein B, low-density lipoprotein cholesterol, and the combination of total and high-density lipoprotein cholesterol. CONCLUSIONS: Changes in lipid levels can explain all or most of the observed benefit of pravastatin. Some treatment effect may also be mediated through nonlipid changes. PMID- 11889010 TI - Sex differences in hospital mortality after coronary artery bypass surgery: evidence for a higher mortality in younger women. AB - BACKGROUND: Data are conflicting over whether women have higher mortality than men after coronary artery bypass graft (CABG) surgery. Younger but not older women hospitalized for acute myocardial infarction have higher in-hospital mortality rates than men. We hypothesized that younger women also have higher in hospital mortality rates after CABG. METHODS AND RESULTS: We studied 51 187 patients (30% women) included in the National Cardiovascular Network database who received CABG at 23 clinical centers between October 1993 and December 1999. Compared with men, fewer women were white and more women had risk factors and comorbidities. These differences were more apparent in younger patients. In all age groups, however, women had higher left ventricular ejection fraction and fewer diseased vessels. Women had higher in-hospital mortality rates than men, but sex differences in mortality were more marked among younger patients. Women <50 years of age were 3 times more likely to die than men (3.4% versus 1.1%), and women 50 to 59 years of age were 2.4 times more likely to die than men (2.6% versus 1.1%). In the older age categories, the sex difference in in-hospital mortality was less marked (P<0.001 for the interaction between sex and age). Adjustment for preoperative risk factors only slightly decreased the strength of this interaction. CONCLUSIONS: Younger women undergoing CABG surgery are at a higher risk of in-hospital death than men, but this difference in risk decreases with advancing age. Additional investigation is needed to determine why in hospital mortality is higher in women after CABG, with particular focus on younger women. PMID- 11889009 TI - Functional and biochemical analysis of endothelial (dys)function and NO/cGMP signaling in human blood vessels with and without nitroglycerin pretreatment. AB - BACKGROUND: In experimental animal models, long-term in vivo treatment with nitroglycerin (NTG) induces both endothelial dysfunction and tolerance to nitrates. However, it is still controversial whether nitrate tolerance in humans is associated with both endothelial dysfunction and impaired vascular response to nitrovasodilator-derived NO. METHODS AND RESULTS: Patients undergoing elective bypass surgery were randomized to receive 48 hours of continuous NTG infusion (NTG group) or no nitrate therapy (control group). Segments of surgically removed arteria mammaria, vena saphena, and arteria radialis not required for the bypass procedure were used to examine (1) the vascular responsiveness to NTG and the endothelium-dependent vasodilator acetylcholine; (2) the expression of the NO target, the soluble guanylyl cyclase; (3) the expression of the soluble guanylyl cyclase/cGMP effector target, the cGMP-dependent protein kinase (cGK); and (4) the cGK activity as assessed by the phosphorylation state of its vascular substrate, the vasodilator-stimulated phosphoprotein at serine(239) (P-VASP). NTG treatment caused a marked degree of nitrate tolerance in all 3 vessel types studied and a significant cross-tolerance to the endothelium-dependent vasodilator acetylcholine in A. mammaria and A. radialis. Although soluble guanylyl cyclase, cGK-I, and VASP expression levels were not modified by NTG treatment, a marked decrease of P-VASP, a surrogate parameter for in-vivo cGK-I activity, was observed. CONCLUSIONS: We conclude that long-term NTG treatment induces endothelial dysfunction and impaired vascular NO/cGMP signaling in humans, which can be monitored by measuring P-VASP levels. PMID- 11889011 TI - Tolerability of beta-blocker initiation and titration in the Metoprolol CR/XL Randomized Intervention Trial in Congestive Heart Failure (MERIT-HF). AB - BACKGROUND: beta-Blockade improves survival when administered over a long period of time to patients with heart failure. However, the time course of any possible deterioration during the titration phase has not been reported. METHODS AND RESULTS: We looked at evidence of clinical deterioration in the Metoprolol CR/XL Randomized Intervention Trial in Congestive Heart Failure (MERIT-HF) by analyzing events and symptoms during the first 90 days. During titration, the Kaplan-Meier curves for the combined end point of all-cause mortality/all-cause hospitalization were similar in all patients randomized, with no significant difference in favor of placebo at any visit or in any of the analyzed subgroups (New York Heart Association class II, III/IV, or III/IV with ejection fraction <0.25, heart rate less-than-or-equal 76 bpm, and systolic blood pressure less than-or-equal 120 mm Hg). The curves started to diverge in favor of beta-blockade after 60 days. Low heart rate was the main factor that limited titration. In New York Heart Association class III/IV, 5.9% of the patients receiving placebo discontinued study medicine during the first 90 days compared with 8.1% of those receiving metoprolol CR/XL (P=0.037 unadjusted, P=NS adjusted); corresponding figures in those with New York Heart Association class III/IV and ejection fraction <0.25 were 7.1% and 8.0% (P=NS). From day 90 until the end of the study, more patients in the placebo group discontinued study medicine in all subgroups. There was no change in diuretic or ACE inhibitor dosing with beta-blocker titration. Most patients reported no change in symptoms of breathlessness or fatigue during the titration phase. CONCLUSIONS: When carefully titrated, metoprolol CR/XL can be given safely to the overwhelming majority of patients with stable mild to moderate heart failure, with minimal side effects or deterioration. PMID- 11889012 TI - Heart failure and ventricular dysfunction in patients with single or systemic right ventricles. AB - BACKGROUND: Previous studies suggested a high incidence of congestive heart failure in patients with single and/or systemic right ventricles. The corresponding risk in an adult population is unknown. METHODS AND RESULTS: A cohort of 188 consecutive adult patients with single or systemic right ventricles was prospectively assessed with gated radionuclide angiography (n=135) or 2D echocardiography (n=188) and followed up clinically. Clinical assessment showed 82.4% of the patients were in New York Heart Association class I or II, 13.3% were in class III, and 4.3% were in class IV. Heart failure occurred in 22.2% of patients with transposition of the great arteries and a Mustard procedure, 32.3% of patients with congenitally corrected transposition of the great arteries, and 40% of Fontan-palliated patients. Symptomatic patients had significantly lower anaerobic thresholds (10.3 +/- 2.8 versus 13.2 +/- 4.8 mL center dot kg(-1) center dot min(-1), P=0.006) and peak (center dot)VO(2) (15.2 +/- 4.8 versus 20.3 +/- 6.8 mL center dot kg(-1) center dot min(-1), P<0.00029). Systemic ventricular ejection fraction in symptomatic versus asymptomatic patients at rest was 34.8 +/ 15.7% versus 46.7 +/- 13.4% (P=0.00001). Mortality was 47.1% among symptomatic patients and 5% among asymptomatic patients at 15.7 years of postoperative follow up. Seven of 12 patients with potentially correctable surgical lesions died or persisted in heart failure despite surgery. Best predictors for mortality were New York Heart Association class, systemic ejection fraction, and age at operation. CONCLUSIONS: Patients with single or systemic right ventricles have significant risk for heart failure accompanied by high mortality. This study suggests the importance of identifying this group of patients who are at risk for heart failure and considering strategies to preserve ventricular function. PMID- 11889013 TI - Progression of systolic abnormalities in patients with "isolated" diastolic heart failure and diastolic dysfunction. AB - BACKGROUND: The definition of diastolic heart failure (DHF) relies on the use of sensitive tools to exclude the presence of systolic dysfunction. The use of ejection fraction (EF) of 50% as the cutoff point may not be adequate to address such a task. We believe that systolic dysfunction is common in DHF. METHODS AND RESULTS: Echocardiography with tissue Doppler imaging was performed in 339 subjects, of whom 92 had systolic heart failure (SHF) (EF<50%), 73 had DHF (EF > or = 50% with diastolic abnormalities on Doppler echocardiography), and 68 had isolated diastolic dysfunction (DD); 106 were normal control subjects. Regional myocardial velocity curves were constructed off-line with the use of a 6-basal, 6 midsegmental model. The peak regional myocardial sustained systolic (S(M)) and early diastolic (E(M)) velocities were significantly lower in patients with SHF, DHF, and DD than in control subjects in almost all the myocardial segments. Likewise, the mean S(M) (SHFor=5 mm) underwent forceps biopsy and complete polypectomy. Patients with fundic gland polyps and polyposis syndrome were not included. Specimens were evaluated by primary and reference pathologists, and the complication rate of gastric polypectomy was also determined. RESULTS: Of the 222 polyps, histological examination of the polypectomy specimens revealed tumour like lesions in 77% (10% focal foveolar hyperplasia, 59% hyperplastic polyps, 4% inflammatory fibroid polyps, 4% other polyps) and neoplasia in 19% (10% tubular adenoma, 2% tubulovillous adenoma, 1% high grade intraepithelial neoplasia, 6% adenocarcinoma). When biopsy results were compared, complete agreement was found in 124 cases (55.8%) and, in an additional 77 cases (34.7%), the clinically important differentiation between tumour-like lesions and neoplasia was possible. However, relevant differences were found by the reference pathologist in six cases (2.7%), the most common reason being failure of biopsy to reveal foci of carcinoma in hyperplastic polyps. Bleeding was observed after polypectomy in 16 patients (7.2%), in 15 of whom it was managed conservatively. CONCLUSIONS: We recommend complete removal by an experienced endoscopist of all epithelial gastric polyps larger than 5 mm after thorough individualised risk-benefit analysis. PMID- 11889064 TI - The menstrual cycle affects rectal sensitivity in patients with irritable bowel syndrome but not healthy volunteers. AB - BACKGROUND: We have previously shown that the menstrual cycle has no effect on rectal sensitivity of normal healthy women, despite them having looser stools at the time of menses. Patients with irritable bowel syndrome (IBS) often report significant exacerbation of their IBS symptoms with menses, raising the possibility that IBS patients may respond differently to the menstrual cycle. AIM AND METHODS: Rectal responses to balloon distension during days 1-4 (menses), 8 10 (follicular phase), 18-20 (luteal phase), and 24-28 (premenstrual phase) of the menstrual cycle were assessed in 29 female IBS patients (aged 21-44 years), diagnosed by the Rome I criteria. During the course of the study patients completed symptom diaries to assess abdominal pain and bloating (visual analogue scale), and frequency and consistency of bowel habits. In addition, levels of anxiety and depression were assessed using the hospital anxiety and depression questionnaire. RESULTS: Menses was associated with a worsening of abdominal pain and bloating compared with most other phases of the menstrual cycle (p<0.05). Bowel habits also became more frequent (p<0.05) and patients tended to have a lower general well being. Rectal sensitivity increased at menses compared with all other phases of the cycle (p<0.05). There was no associated change in rectal compliance, wall tension, or motility index. Neither was there any difference in resting anal pressure or the distension volumes required to relax the internal anal sphincter during the menstrual cycle. CONCLUSION: These data (1) confirm that IBS symptomatology is exacerbated at menses and (2) show for the first time that in contrast with healthy women, rectal sensitivity changes with the menstrual cycle. These cyclical changes in sensitivity suggest that women with IBS respond differently to fluctuations in their sex hormonal environment or its consequences compared with healthy females. PMID- 11889065 TI - Microprocessor controlled movement of solid colonic content using sequential neural electrical stimulation. AB - BACKGROUND AND AIMS: Invoked peristaltic contractions and movement of solid content have not been attempted in normal canine colon. The purpose of this study was to determine if movement of solid content through the colon could be produced by microprocessor controlled sequential stimulation. METHODS: The study was performed on six anaesthetised dogs. At laparotomy, a 15 cm segment of descending colon was selected, the proximal end closed with a purse string suture, and the distal end opened into a collecting container. Four sets of subserosal stimulating electrodes were implanted at 3 cm intervals. The segment of bowel was filled with a mixture of dog food and 50 plastic pellets before each of 2-5 random sessions of non-stimulated or stimulated emptying. Propagated contractions were generated using microprocessor controlled bipolar trains of 50 Hz rectangular voltage having 20 V (peak to peak) amplitude, 18 second stimulus duration, and a nine second phase lag between stimulation trains in sequential electrode sets. RESULTS: Electrical stimulation using the above mentioned parameters resulted in powerful phasic contractions that closed the lumen. By phase locking the stimulation voltage between adjacent sets of electrodes, propagated contractions could be produced in an aboral or orad direction. The number of evacuated pellets during the stimulation sessions was significantly higher than during the non-stimulated sessions (p<0.01). CONCLUSIONS: Microprocessor controlled electrical stimulation accelerated movement of colonic content suggesting the possibility of future implantable colonic stimulators. PMID- 11889066 TI - Prevalence of faecal incontinence in adults aged 40 years or more living in the community. AB - BACKGROUND: Prevalence studies of faecal incontinence in the general population are rare and the impact of faecal incontinence on quality of life has not been previously addressed. AIMS: To establish the prevalence of faecal incontinence in adults in terms of frequency of leakage, degree of soiling, and level of impact on quality of life. METHODS: In a cross sectional postal survey, 15 904 adults aged 40 years or more (excluding residents of nursing and residential homes) were selected randomly by household from the Leicestershire Health Authority patient register. Participants were asked to complete a confidential health questionnaire. Major faecal incontinence was defined as soiling of underwear or worse with a frequency of several times a month or more. Respondents were also asked if bowel symptoms had an impact on their quality of life. RESULTS: From a total sample of 10 116 respondents, 1.4% reported major faecal incontinence and 0.7% major faecal incontinence with bowel symptoms that had an impact on quality of life. Major faecal incontinence was significantly associated with a lot of impact on quality of life (odds ratio 12.4, 95% confidence interval 7.5-20.6). Incontinence was more prevalent and more severe in older people but there was no significant difference between men and women. CONCLUSIONS: This study has confirmed that faecal incontinence is a fairly common symptom, particularly in older people. Faecal incontinence in men has received little attention in the past and the results from this study indicate that it is as much of a problem in men as it is in women while the level of unmet need in this group is high. Estimates of need for health care for this symptom should be multidimensional and assess both the severity of symptoms and the impact it has on quality of life. PMID- 11889067 TI - The efficacy of azathioprine for the treatment of inflammatory bowel disease: a 30 year review. AB - BACKGROUND: There are limited data on factors predicting response to azathioprine and uncertainty regarding the optimal duration of treatment. PATIENTS AND METHODS: The notes of patients attending the Oxford IBD clinic from 1968 to 1999 were reviewed. Remission was defined as no need for oral steroids for at least three months and relapse was defined as active disease requiring steroids. RESULTS: A total of 622 of 2205 patients were treated with azathioprine (272 Crohn's disease, 346 ulcerative colitis, and four indeterminate colitis). Mean duration of the initial course of treatment was 634 days. The overall remission rates were 45% for Crohn's disease and 58% for ulcerative colitis. For the 424 patients who received more than six months of treatment, remission rates were 64% and 87%, respectively. Factors favouring remission were ulcerative colitis (p=0.0001), lower white blood cell (WBC) or neutrophil count (p=0.0001), higher mean cell volume (p=0.0001), and older age (p=0.05). For Crohn's disease, colonic disease favoured remission (p=0.03). Factors that were not significant were age, sex, lymphocyte count, and dose (mg/kg). The proportion of patients remaining in remission at one, three, and five years was 0.95, 0.69, and 0.55, respectively. The chance of remaining in remission was higher if WBC was less than 5 x 10(9) (p=0.03) and in male patients (p=0.01; Crohn's disease only). There was no difference in relapse rates between Crohn's disease and ulcerative colitis. After stopping azathioprine, the proportion of patients remaining in remission at one, three, and five years was 0.63, 0.44, and 0.35 (222 patients). Duration of azathioprine treatment did not affect the relapse rate after stopping treatment (p=0.68). CONCLUSIONS: Azathioprine is effective treatment for ulcerative colitis and Crohn's disease. The efficacy of azathioprine is reasonably well sustained over five years. PMID- 11889068 TI - Bowel ultrasound in assessment of Crohn's disease and detection of related small bowel strictures: a prospective comparative study versus x ray and intraoperative findings. AB - BACKGROUND AND AIM: Despite the fact that bowel ultrasound (US) has recently been proved to be useful in the assessment of bowel diseases, uncertainty persists as to its diagnostic role in patients with complicated Crohn's disease (CD). Therefore, we have prospectively investigated the accuracy of US compared with x ray procedures and intraoperative findings in detecting small bowel strictures complicating CD as well as its reliability in assessing disease extent and location within the bowel. METHODS: A series of 296 consecutive patients with proven CD admitted to L Sacco University Hospital between 1997 and 1999, having undergone complete radiographic evaluation (including small bowel x ray, colonoscopy, or double contrast barium enema), were enrolled in the study. Bowel US was performed in each patient by two experienced operators unaware of the results of other diagnostic procedures. The accuracy of US in detecting strictures compared with x ray studies was determined separately in two different groups of patients: 211 patients treated conservatively (non-operative CD) and 85 patients who were candidates for surgery for CD complications or unresponsiveness to medical therapy (operative CD). RESULTS: Overall sensitivity and specificity of US in assessing the anatomical distribution of CD were 93% and 97%, respectively. The extent of ileal disease measured at US correlated well with that determined by x ray (r=0.52, p<0.001) in medically treated patients as well as with that measured intraoperatively in surgical patients (r=0.64, p<0.001). One or more stenoses were detected in 75 patients (35.5%) at small bowel enteroclysis in the non-operative CD group compared with 70 (82%) in the operative CD series. Sensitivity, specificity, and positive predictive values of bowel US in the detection of strictures were 79%, 98%, and 95% in non-operative CD patients and 90%, 100%, and 100% in operative CD cases, respectively. CONCLUSIONS: In experienced hands, bowel US is an accurate technique for assessing CD extent and location and is very helpful in detecting small bowel strictures, especially in very severe cases that are candidates for surgery. The use of bowel US is therefore justified as a primary investigation in CD patients in whom complications are suspected. PMID- 11889069 TI - The vascularity of internal fistulae in Crohn's disease: an in vivo power Doppler ultrasonography assessment. AB - AIM: To evaluate the efficacy of power Doppler sonography (US) in depicting internal fistulae and their vascularity, and to study the characteristics of blood flow within the fistula wall. PATIENTS AND METHODS: The study involved 45 consecutive patients with Crohn's disease and suspected internal fistulae detected by grey scale US. The fistulae were subsequently evaluated using power Doppler US to reveal any areas of increased vascularity, and the results were compared with radiographic, endoscopic, or intraoperative findings. Whenever feasible, we also performed spectral analysis of blood flow revealed by power Doppler US, calculated its resistance index (RI), and analysed its characteristics, reproducibility, and relationship with biochemical and clinical variables (Crohn's disease activity index, disease duration, location, and abdominal complications). RESULTS: Power Doppler US revealed vascularity in all of the internal fistulae that where subsequently confirmed by diagnostic procedures. In the case of intra-abdominal abscesses in the vicinity of the fistula, vascular signals were detected mostly around and not within the lesions. The intensity and distribution of the signals differed within the fistulae tracks and had only slight day to day reproducibility; furthermore, there was no significant correlation with clinical or biochemical variables. Spectral analyses of blood flow within the fistulae revealed arterial flow in 96.7% of patients (median RI 0.715). RI was a more reproducible parameter and significantly correlated with clinical (r= 0.54) and biochemical activity (r= 0.56) of CD. It was also higher in fistulae complicated by abscesses. CONCLUSION: Power Doppler US can reveal the presence of vasculature within the wall of internal fistulae and therefore enhance grey scale US performance. The RI characteristics of blood flow within the fistulae are reproducible and correlate with biochemical and clinical disease activity. PMID- 11889070 TI - Human neutrophil lipocalin is a unique marker of neutrophil inflammation in ulcerative colitis and proctitis. AB - BACKGROUND AND AIM: Accumulation and infiltration by neutrophil granulocytes is a prominent feature in the local inflammatory process in ulcerative colitis (UC). The present study was performed to evaluate human neutrophil lipocalin (HNL) as a specific neutrophil marker in the inflamed lesions of the colon and rectum in patients with colitis and proctitis. METHODS: The activity of intestinal neutrophils with respect to release of granule proteins was studied in 18 patients with UC (10 with colitis and eight with isolated proctitis) and in 18 healthy controls using perfusion fluid and biopsies from the sigmoid colon and rectum. The released amounts of the neutrophil granule proteins HNL and myeloperoxidase (MPO) were determined by radioimmunoassays, and the location of HNL and MPO in biopsies from colonic mucosa was examined by immunohistochemistry. RESULTS: Mucosal release of HNL and MPO was increased 10-55-fold in patients with colitis and proctitis compared with controls. Their bowel biopsies demonstrated that only neutrophils were stained with anti-HNL. We also found correlations between HNL and levels of granulocyte/macrophage-colony stimulating factor (GM CSF) and interleukin 8 (IL-8) in perfusion fluids from the sigmoidal segments of patients with proctitis, between HNL and GM-CSF in rectal segments in patients with proctitis, and in sigmoidal segments in patients with colitis. CONCLUSION: We conclude that the increased release of HNL and MPO in colorectal perfusion fluids indicates neutrophil involvement in the local inflammatory process, and suggest that HNL may serve as a specific marker of intestinal neutrophil activation in UC. GM-CSF, and to some extent IL-8, may play a role in neutrophil accumulation and priming in this disease. PMID- 11889072 TI - Aberrant P-cadherin expression is an early event in hyperplastic and dysplastic transformation in the colon. AB - BACKGROUND: Colorectal adenomatous and, probably, hyperplastic polyp development requires epithelial remodelling and stratification, with loss of E-cadherin expression implicated in adenoma formation. We have shown that P-cadherin, normally expressed in stratified epithelia and placenta, is aberrantly expressed in disturbed epithelial architecture associated with colitis. AIMS: (i) To investigate the role of P-cadherin in colonic polyp formation. (ii) To ascertain whether expression of P-cadherin is independent of or correlated with expression of its associated proteins--E-cadherin, beta-catenin, and gamma-catenin. (iii) To determine if P-cadherin is functional regarding catenin binding in polyps. METHODS: Expression and localisation of cadherins (E- and P-) and their associated catenins (beta- and gamma-) were determined in aberrant crypt foci (ACF), in polyps with hyperplastic morphology (hyperplastic polyps and serrated adenomas), and in adenomatous polyps by immunohistochemistry, western blotting, and mRNA in situ hybridisation. Assessment of cadherin-catenin binding was evaluated by co-immunoprecipitation. Adenomatous polyposis coli (APC) mutation was assessed in adenomatous polyps. RESULTS: P-cadherin was expressed from ACF through to hyperplastic and adenomatous polyps. Alterations in E-cadherin and catenin expression occurred later, with variant patterns in (i) ACF, (ii) hyperplastic polyps and serrated adenomas, and (iii) adenomatous polyps. P cadherin present in adenomas was functional with regard to catenin binding, and its expression was independent of APC mutational status. CONCLUSIONS: P-cadherin is aberrantly expressed from the earliest morphologically identifiable stage of colonocyte transformation, prior to changes in E-cadherin, catenin, and APC expression/mutation. P-cadherin expression alone does not predict tissue morphology, and such expression is independent of that of associated cadherins and catenins. PMID- 11889073 TI - A polymorphism in the methylenetetrahydrofolate reductase gene predisposes to colorectal cancers with microsatellite instability. AB - BACKGROUND: The enzyme methylenetetrahydrofolate reductase (MTHFR) catalyses the formation of folate intermediates that are vital to methylation reactions. A polymorphic variant (TT) has been linked to reduced levels of plasma folate, aberrant DNA methylation in leucocytes, and increased risk of colorectal cancer (CRC) under conditions of low folate intake. The cystathionine beta-synthase (CBS) enzyme reduces homocysteine levels and thus may protect against CRC. The CBS gene has a variant, 844ins68, that has been linked with increased activity. These variants may be involved in the development of the subgroup of CRC displaying aberrant DNA methylation and frequently associated with microsatellite instability (MSI). AIM: To investigate the frequencies of the TT and 844ins68 genotypes in CRC patients with MSI+ tumours compared with those with MSI- tumours and a control population. SUBJECTS: Patients with CRC (n=501) and healthy control subjects (n=1207) were studied. CRC cases were classified as MSI+ (n=75) or MSI- (n=426) based on deletions within the BAT-26 mononucleotide repeat. METHODS: Subjects were genotyped for MTHFR using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and PCR-restriction fragment length polymorphism (PCR-RFLP) techniques, and for CBS using PCR. RESULTS: The MTHFR TT genotype was more frequent in older CRC patients (>or=70 y) compared with equivalent aged controls (p=0.03), was associated with a significantly later age of diagnosis in patients with proximal colon tumours (p=0.02), and was almost twice as frequent in MSI+ than in MSI- tumours (p=0.05). Compared with normal controls, the 844ins68 variant of CBS was less frequent in patients with proximal tumours (p=0.02). CONCLUSIONS: The TT genotype of MTHFR is associated with an increased risk of CRC in older populations, possibly due to age related disturbances in folate metabolism. The TT genotype appears to predispose to CRC that is MSI+. This may reflect the involvement of aberrant DNA methylation frequently associated with MSI+. The 844ins68 CBS polymorphism may protect against proximal tumours. PMID- 11889071 TI - Dichotomal role of inhibition of p38 MAPK with SB 203580 in experimental colitis. AB - BACKGROUND: Crohn's disease is characterised by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important perpetuating role. Mitogen activated protein kinase p38 (p38 MAPK) has been established as a major regulator of the inflammatory response, especially with regard to production of proinflammatory cytokines, but its role in inflammatory bowel disease is unexplored. In this paper we describe the effects of a specific p38 MAPK inhibitor, SB 203580, in trinitrobenzene sulphonic acid (TNBS) induced colitis in mice. RESULTS: SB 203580 had a dichotomal effect in TNBS mice. Weight loss of TNBS mice treated with SB 203580 was significantly worse and colon weight on sacrifice was significantly increased in MAPK inhibitor treated TNBS mice (229.2 mg and 289.1 mg, respectively). However, the total number of cells in the caudal lymph node decreased to 188.8 x 10(4) cells in SB 203580 treated TNBS mice compared with 334 x 10(4) cells in vehicle treated mice. CD3/CD28 double stimulated caudal lymph node cells of SB 203580 treated mice showed decreased interferon gamma production but increased tumour necrosis factor alpha production. The concentration of interleukin 12p70 in colon homogenates was significantly decreased in SB 203580 treated mice whereas concentrations of interleukin 12p40, tumour necrosis factor alpha, and interleukin 10 were similar in vehicle and SB 203580 treated TNBS mice. CONCLUSION: Our results reveal a dichotomy in p38 MAPK action during experimental colitis. PMID- 11889075 TI - Detection of epithelial tumour RNA in the plasma of colon cancer patients is associated with advanced stages and circulating tumour cells. AB - BACKGROUND: Although circulating tumour DNA has been detected in patients with different types of cancer, little is known of free RNA in cancer patients. AIMS: We investigated the presence of RNA from epithelial tumours in plasma from patients with colorectal carcinomas, and its correlation with tumour characteristics and circulating tumour cells. METHODS: beta-actin mRNA was analysed to assess the viability of plasma RNA in samples from 53 patients with colonic cancer and 25 controls. Subsequently, nested primers were used to detect the presence of cytokeratin 19 (CK19) and carcinoembryonic antigen (CEA) RNA in the same samples. Nine clinicopathological parameters were studied to correlate the molecular and clinical parameters. Additionally, we investigated for micrometastases in blood in 18 of these patients and in 10 of the controls samples. RESULTS: All samples had detectable quantities of beta-actin RNA. In the controls, one case (4%) was positive for CEA and five (20%) for CK19 RNA; of the 53 patients, 17 cases (32%) were positive for CEA and 39 (73.6%) for CK19 RNA. This was statistically significant (p=0.000001). Advanced stages (p=0.03) and soluble CEA status (p=0.03) were associated with the presence of CEA, CK19, or both RNAs in plasma. Lymph node metastases (p=0.06) and vascular invasion (p=0.07) were almost significant. On the basis of these results, we examined the possible presence of micrometastases in blood in several of these patients. The presence of plasma tumour RNA was found to be associated with circulating tumour cells in blood (p=0.04). CONCLUSIONS: Epithelial tumour RNA is detectable in plasma from colon cancer patients. This molecular event is associated with advanced stages and circulating tumour cells. Our results could offer new approaches in the diagnosis and monitoring of colon cancer. PMID- 11889074 TI - Stimulation of the intestinal Cdx2 homeobox gene by butyrate in colon cancer cells. AB - BACKGROUND: The transcription factor encoded by the intestinal Cdx2 homeobox gene and treatment with sodium butyrate (NaB), a byproduct of fibre fermentation by colonic bacteria, exert similar effects on colon cancer cell lines as they both inhibit cell growth and stimulate cell differentiation and apoptosis. AIM: To investigate whether NaB regulates expression of the Cdx2 gene in colon cancer cell lines. METHODS: Human adenocarcinoma cell lines Caco2 and HT29 were grown in the presence or absence of NaB. Cells were analysed for Cdx2 mRNA expression by reverse transcription-polymerase chain reaction, for protein expression by western blotting and electromobility shift assays, and for transcriptional activity of the Cdx2 promoter by transfection with luciferase reporter plasmids. RESULTS: In HT29 and Caco2 cells, NaB stimulated Cdx2 mRNA and protein expression as well as transcriptional activity of the Cdx2 promoter. Stimulation of the activity of the Cdx2 promoter by NaB was dose and time dependent. The Cdx2 promoter contains discrete regions that participate in or inversely that blunt the stimulatory effect exerted by NaB. In addition, NaB stimulated the transcriptional activity of the Cdx2 promoter downregulated by oncogenic ras. CONCLUSION: This study is the first report of an intestine specific transcription factor, Cdx2, stimulated by butyrate. Thus it provides a new mechanism whereby butyrate controls proliferation and differentiation of colon cancer cells. PMID- 11889076 TI - Pancreatic stellate cells respond to inflammatory cytokines: potential role in chronic pancreatitis. AB - BACKGROUND: It is now generally accepted that chronic pancreatic injury and fibrosis may result from repeated episodes of acute pancreatic necroinflammation (the necrosis-fibrosis sequence). Recent studies suggest that pancreatic stellate cells (PSCs), when activated, may play an important role in the development of pancreatic fibrosis. Factors that may influence PSC activation during pancreatic necroinflammation include cytokines known to be important in the pathogenesis of acute pancreatitis, such as tumour necrosis factor alpha (TNF-alpha), and the interleukins 1, 6, and 10 (IL-1, IL-6, and IL-10). AIM: To determine the effects of these cytokines on PSC activation, as assessed by cell proliferation, alpha smooth muscle actin (alpha-SMA) expression, and collagen synthesis. METHODS: Cultured rat PSCs were incubated with cytokines for 24 hours. Cell proliferation was assessed by measuring (3)H thymidine incorporation into cellular DNA, alpha SMA expression by western blotting, and collagen synthesis by incorporation of (14)C proline into collagenase sensitive protein. mRNA levels for procollagen alpha(1)(1) in PSCs were determined by northern and dot blotting methods. RESULTS: Expression of alpha-SMA by PSCs was increased on exposure to each of the cytokines used in the study. Stellate cell proliferation was stimulated by TNF alpha but inhibited by IL-6, while IL-1 and IL-10 had no effect on PSC proliferation. Collagen synthesis by PSCs was stimulated by TNF-alpha and IL-10, inhibited in response to IL-6, and unaltered by IL-1. Changes in collagen protein synthesis in response to TNF-alpha, IL-10, and IL-6 were not regulated at the mRNA level in the cells. CONCLUSION: This study has demonstrated that PSCs have the capacity to respond to cytokines known to be upregulated during acute pancreatitis. Persistent activation of PSCs by cytokines during acute pancreatitis may be a factor involved in the progression from acute pancreatitis to chronic pancreatic injury and fibrosis. PMID- 11889077 TI - Apoptosis and proliferation of acinar and islet cells in chronic pancreatitis: evidence for differential cell loss mediating preservation of islet function. AB - BACKGROUND: Chronic pancreatitis is characterised clinically by early exocrine insufficiency, with diabetes mellitus occurring as a late phenomenon. This is mirrored pathologically by extensive acinar cell destruction and islet preservation. The mechanisms underlying this differential rate of cellular destruction are unknown. AIMS: To test the hypothesis that acinar loss and islet preservation in chronic pancreatitis occurs due to differential epithelial kinetics and investigate the role of inflammatory cells and cell cycle associated molecules. METHODS: Archival tissue from six chronic pancreatitis cases was compared with six normal controls using TUNEL and immunohistochemistry for CD3, CD20, CD68, MIB-1, Bcl-2, Bax, Fas, Fas ligand, retinoblastoma protein (Rb), and tissue inhibitor of metalloproteinases 1 (TIMP-1) and 2 (TIMP-2). RESULTS: The acinar cell apoptotic index (AI) and proliferation index were higher in chronic pancreatitis than controls. T lymphocytes diffusely infiltrated fibrous bands and acini but rarely islets. Acinar Bcl-2 expression exceeded islet expression in chronic pancreatitis and controls while Bax was strongly expressed by a subset of islet cells and weakly by centroacinar cells. Islet Fas and Fas ligand expression exceeded acinar expression in chronic pancreatitis and controls. Acinar Rb expression was higher in chronic pancreatitis than in controls. Islets in chronic pancreatitis and controls showed intense TIMP-1 and TIMP-2 expression. CONCLUSION: Apoptosis plays a significant role in acinar loss in chronic pancreatitis. Acinar Bcl-2 and islet Bax expression indicates complex AI control. Increased acinar Rb expression in chronic pancreatitis may differentially promote acinar loss. Fas ligand expression may be restricted to islet cell membranes through TIMP-1 expression and inhibit islet damage by promoting apoptosis of cytotoxic T lymphocytes. PMID- 11889078 TI - Photodynamic therapy for cancer of the pancreas. AB - BACKGROUND: Few pancreatic cancers are suitable for surgery and few respond to chemoradiation. Photodynamic therapy produces local necrosis of tissue with light after prior administration of a photosensitising agent, and in experimental studies can be tolerated by the pancreas and surrounding normal tissue. AIMS: To undertake a phase I study of photodynamic therapy for cancer of the pancreas. PATIENTS: Sixteen patients with inoperable adenocarcinomas (2.5-6 cm in diameter) localised to the region of the head of the pancreas were studied. All presented with obstructive jaundice which was relieved by biliary stenting prior to further treatment. METHODS: Patients were photosensitised with 0.15 mg/kg meso tetrahydroxyphenyl chlorin intravenously. Three days later, light was delivered to the cancer percutaneously using fibres positioned under computerised tomographic guidance. Three had subsequent chemotherapy. RESULTS: All patients had substantial tumour necrosis on scans after treatment. Fourteen of 16 left hospital within 10 days. Eleven had a Karnofsky performance status of 100 prior to treatment. In 10 it returned to 100 at one month. Two patients with tumour involving the gastroduodenal artery had significant gastrointestinal bleeds (controlled without surgery). Three patients developed duodenal obstruction during follow up that may have been related to treatment. There was no treatment related mortality. The median survival time after photodynamic therapy was 9.5 months (range 4-30). Seven of 16 patients (44%) were alive one year after photodynamic therapy. CONCLUSIONS: Photodynamic therapy can produce necrosis in pancreatic cancers with an acceptable morbidity although care is required for tumours invading the duodenal wall or involving the gastroduodenal artery. Further studies are indicated to assess its influence on the course of the disease, alone or in combination with chemoradiation. PMID- 11889079 TI - Attenuated acute liver injury in mice by naked hepatocyte growth factor gene transfer into skeletal muscle with electroporation. AB - BACKGROUND: Hepatocyte growth factor (HGF) plays an essential role in hepatic development and regeneration, and shows proliferative and antiapoptotic activity in hepatocytes. AIMS: To establish an effective new method for HGF gene transfer in vivo and to investigate its effects in acute experimental liver injury. ANIMALS: Eight week old female mice were used. METHODS: Rat HGF gene in a modified pKSCX plasmid was transferred to the tibialis anterior muscle by electroporation using a pulse generator. Four days later, plasma HGF concentrations were determined by enzyme linked immunosorbent assay every two days for three weeks. To confirm the efficacy of electroporation, a plasmid bearing green fluorescence protein (GFP) was transferred similarly. Four days after electroporation, carbon tetrachloride (CCl(4)) was administered to mice to induce acute liver injury. Plasma alanine aminotransferase (ALT) activity was measured. Hepatic apoptosis was assessed by Hoechst 33258 staining and the TUNEL method. RESULTS: Fluorescence microscopy showed strong green fluorescence where the GFP gene had been transferred into muscle. In mice given the HGF gene, HGF in plasma was increased up to fourfold from pretreatment amounts, peaking 6-9 days after electroporation and quickly decreasing within three weeks. Compared with the group without HGF transfer, the percentage of apoptotic hepatocytes after CCl(4) intoxication was significantly lower, as was ALT activity. In addition, ALT activity normalised more rapidly in the HGF gene transfer group. CONCLUSIONS: Naked DNA injection and transfer by electroporation efficiently brings about HGF expression in vivo, which can attenuate acute liver injury. PMID- 11889080 TI - A ligand for peroxisome proliferator activated receptor gamma inhibits cell growth and induces apoptosis in human liver cancer cells. AB - BACKGROUND AND AIMS: Induction of apoptosis of cancer cells through ligands of nuclear hormone receptors (NHRs) is a new approach in cancer therapy. Recently, one of the NHRs, peroxisome proliferator activated receptor gamma (PPARgamma), has been shown to influence cell growth in certain cancer cells although its effect on hepatocellular carcinoma (HCC) has not been analysed. METHODS: Experiments were conducted using three human liver cancer cell lines, PLC/PRF/5, Hep G2 and HuH-7, in vitro. These cells were exposed to troglitazone, a synthetic ligand for PPARgamma, and the effects on cell growth were analysed. RESULTS: Expression of PPARgamma mRNA was detected in all three liver cancer cell lines. Activation of PPARgamma by troglitazone caused a marked growth inhibition in a dose dependent manner in three hepatoma cell lines. The DNA fragmentation ELISA assay and Hoechst 33258 staining revealed that the growth inhibitory effect by adding troglitazone was due to apoptosis of PLC/PRF/5, which strongly expressed PPARgamma. Troglitazone also induced activation of the cell death protease, caspase 3, but not caspase 8, in PLC/PRF/5 cells. However, expression levels of antiapoptotic factor bcl-2 and apoptosis inducing factor bax were not affected. CONCLUSION: Our study showed that PPARgamma was expressed in human liver cancer cells and that the ligand for PPARgamma, troglitazone, inhibited the growth of these cells by inducing apoptosis through caspase 3 activation, indicating that troglitazone could be potentially useful as an apoptosis inducer for the treatment of HCC. PMID- 11889081 TI - Hospital admissions for peptic ulcer and indigestion in London and New York in the 19th and early 20th centuries. AB - The occurrence of peptic ulcer increased rapidly in all Western countries from the 19th to the 20th century, attributed to a possible epidemic of Helicobacter pylori, a new pathogenic strain, or a change in host susceptibility. The early trends in hospital admissions for peptic ulcer and dyspepsia in London and New York during the 19th century are reviewed to test these hypotheses. PMID- 11889083 TI - Recurrent diarrhoea and weight loss associated with cessation of smoking in undiagnosed coeliac disease. PMID- 11889084 TI - Forcing mycoplasma mobile into line. PMID- 11889085 TI - Plants in the pink: cytokinin production by methylobacterium. PMID- 11889082 TI - Hepatic stellate cells: role in microcirculation and pathophysiology of portal hypertension. AB - Accumulating evidence suggests that stellate cells are involved in the regulation of the liver microcirculation and portal hypertension. Activated hepatic stellate cells have the necessary machinery to contract or relax in response to a number of vasoactive substances. Because stellate cells play a role in both fibrosis and portal hypertension, they are currently regarded as therapeutic targets to prevent and treat the complications of chronic liver disease. PMID- 11889086 TI - Modularity and specialization in superfamily 1 and 2 helicases. PMID- 11889087 TI - Force and velocity of mycoplasma mobile gliding. AB - The effects of temperature and force on the gliding speed of Mycoplasma mobile were examined. Gliding speed increased linearly as a function of temperature from 0.46 microm/s at 11.5 degrees C to 4.0 microm/s at 36.5 degrees C. A polystyrene bead was attached to the tail of M. mobile using a polyclonal antibody raised against whole M. mobile cells. Cells attached to beads glided at the same speed as cells without beads. When liquid flow was applied in a flow chamber, cells reoriented and moved upstream with reduced speeds. Forces generated by cells at various gliding speeds were calculated by multiplying their estimated frictional drag coefficients with their velocities relative to the liquid. The gliding speed decreased linearly with force. At zero speed, the force measurements extrapolated to 26 pN at 22.5 and 27.5 degrees C. At zero force, the speed extrapolated to 2.3 and 3.3 microm/s at 22.5 and 27.5 degrees C, respectively--the same speeds as those observed for free gliding cells. Cells attached to beads were also trapped by an optical tweezer, and the stall force was measured to be 26 to 28 pN (17.5 to 27.5 degrees C). The gliding speed depended on temperature, but the maximum force did not, suggesting that the mechanism is composed of at least two steps, one that generates force and another that allows displacement. Other implications of these results are discussed. PMID- 11889088 TI - tRNA is the source of low-level trans-zeatin production in Methylobacterium spp. AB - Pink-pigmented facultatively methylotrophic bacteria (PPFMs), classified as Methylobacterium spp., are persistent colonizers of plant leaf surfaces. Reports of PPFM-plant dialogue led us to examine cytokinin production by PPFMs. Using immunoaffinity and high-performance liquid chromatography (HPLC) purification, we obtained 22 to 111 ng of trans-zeatin per liter from culture filtrates of four PPFM leaf isolates (from Arabidopsis, barley, maize, and soybean) and of a Methylobacterium extorquens type culture originally recovered as a soil isolate. We identified the zeatin isolated as the trans isomer by HPLC and by a radioimmunoassay in which monoclonal antibodies specific for trans-hydroxylated cytokinins were used. Smaller and variable amounts of trans-zeatin riboside were also recovered. trans-Zeatin was recovered from tRNA hydrolysates in addition to the culture filtrates, suggesting that secreted trans-zeatin resulted from tRNA turnover rather than from de novo synthesis. The product of the miaA gene is responsible for isopentenylation of a specific adenine in some tRNAs. To confirm that the secreted zeatin originated from tRNA, we mutated the miaA gene of M. extorquens by single exchange of an internal miaA fragment into the chromosomal gene. Mutant exconjugants, confirmed by PCR, did not contain zeatin in their tRNAs and did not secrete zeatin into the medium, findings which are consistent with the hypothesis that all zeatin is tRNA derived rather than synthesized de novo. In germination studies performed with heat-treated soybean seeds, cytokinin null (miaA) mutants stimulated germination as well as wild-type bacteria. While cytokinin production may play a role in the plant-PPFM interaction, it is not responsible for stimulation of germination by PPFMs. PMID- 11889089 TI - Effect of different concentrations of H-NS protein on chromosome replication and the cell cycle in Escherichia coli. AB - Flow cytometric analysis showed that the hns205 and hns206 mutants, lacking the abundant nucleoid-associated protein H-NS, have decreased origin concentration, as well as a low number of origins per cell (ploidy). The most striking observation was that the low ploidy was due to a very short replication time, e.g., at 30 degrees C it was halved compared to that of the hns(+) strain. The decreased origin concentration was not caused by a decreased dnaA gene expression, and the hns206 mutant had normal DnaA protein concentrations. The replication phenotypes of the hns206 mutant were independent of RpoS. Cells overproducing H-NS from a LacI-controlled plasmid had a normal origin concentration, indicating that H-NS is not controlling initiation. A wild-type H NS concentration is, however, required to obtain a wild-type origin concentration, since cells with an intermediate H-NS concentration had an intermediate origin concentration. Two lines of evidence point to an indirect effect of H-NS on initiation. First, H-NS did not show high-affinity binding to any part of oriC, and H-NS had no effect on transcription entering oriC from the mioC promoter. Second, in a shift experiment with the hns206 mutant, when H-NS protein was induced to wild-type levels within 10 min, it took more than one generation before the origin concentration started to increase. PMID- 11889090 TI - Characterization of an ADP-ribosyltransferase toxin (AexT) from Aeromonas salmonicida subsp. salmonicida. AB - An ADP-ribosylating toxin named Aeromonas salmonicida exoenzyme T (AexT) in A. salmonicida subsp. salmonicida, the etiological agent of furunculosis in fish, was characterized. Gene aexT, encoding toxin AexT, was cloned and characterized by sequence analysis. AexT shows significant sequence similarity to the ExoS and ExoT exotoxins of Pseudomonas aeruginosa and to the YopE cytotoxin of different Yersinia species. The aexT gene was detected in all of the 12 A. salmonicida subsp. salmonicida strains tested but was absent from all other Aeromonas species. Recombinant AexT produced in Escherichia coli possesses enzymatic ADP ribosyltransferase activity. Monospecific polyclonal antibodies directed against purified recombinant AexT detected the toxin produced by A. salmonicida subsp. salmonicida and cross-reacted with ExoS and ExoT of P. aeruginosa. AexT toxin could be detected in a wild type (wt) strain of A. salmonicida subsp. salmonicida freshly isolated from a fish with furunculosis; however, its expression required contact with RTG-2 rainbow trout gonad cells. Under these conditions, the AexT protein was found to be intracellular or tightly cell associated. No AexT was found when A. salmonicida subsp. salmonicida was incubated in cell culture medium in the absence of RTG-2 cells. Upon infection with wt A. salmonicida subsp. salmonicida, the fish gonad RTG-2 cells rapidly underwent significant morphological changes. These changes were demonstrated to constitute cell rounding, which accompanied induction of production of AexT and which led to cell lysis after extended incubation. An aexT mutant which was constructed from the wt strain with an insertionally inactivated aexT gene by allelic exchange had no toxic effect on RTG-2 cells and was devoid of AexT production. Hence AexT is directly involved in the toxicity of A. salmonicida subsp. salmonicida for RTG-2 fish cells. PMID- 11889091 TI - Construction of an integration-proficient vector based on the site-specific recombination mechanism of enterococcal temperate phage phiFC1. AB - The genome of temperate phage phiFC1 integrates into the chromosome of Enterococcus faecalis KBL 703 via site-specific recombination. In this study, an integration vector containing the attP site and putative integrase gene mj1 of phage phiFC1 was constructed. A 2,744-bp fragment which included the attP site and mj1 was inserted into a pUC19 derivative containing the cat gene to construct pEMJ1-1. E. faecalis KBL 707, which does not contain the bacteriophage but which has a putative attB site within its genome, could be transformed by pEMJ1-1. Southern hybridization, PCR amplification, and DNA sequencing revealed that pEMJ1 1 was integrated specifically at the putative attB site within the E. faecalis KBL 707 chromosome. This observation suggested that the 2,744-bp fragment carrying mj1 and the attP site of phage phiFC1 was sufficient for site-specific recombination and that pEMJ1-1 could be used as a site-specific integration vector. The transformation efficiency of pEMJ1-1 was as high as 6 x 10(3) transformants/microg of DNA. In addition, a vector (pATTB1) containing the 290-bp attB region was constructed. pATTB1 was transformed into Escherichia coli containing a derivative of the pET14b vector carrying attP and mj1. This resulted in the formation of chimeric plasmids by site-specific recombination between the cloned attB and attP sequences. The results indicate that the integration vector system based on the site-specific recombination mechanism of phage phiFC1 can be used for genetic engineering in E. faecalis and in other hosts. PMID- 11889092 TI - Molecular analysis of the gene encoding a novel chitin-binding protease from Alteromonas sp. strain O-7 and its role in the chitinolytic system. AB - Alteromonas sp. strain O-7 secretes several proteins in response to chitin induction. We have found that one of these proteins, designated AprIV, is a novel chitin-binding protease involved in chitinolytic activity. The gene encoding AprIV (aprIV) was cloned in Escherichia coli. DNA sequencing analysis revealed that the open reading frame of aprIV encoded a protein of 547 amino acids with a calculated molecular mass of 57,104 Da. AprIV is a modular enzyme consisting of five domains: the signal sequence, the N-terminal proregion, the family A subtilase region, the polycystic kidney disease domain (PkdD), and the chitin binding domain type 3 (ChtBD3). Expression plasmids coding for PkdD or both PkdD and ChtBD (PkdD-ChtBD) were constructed. The PkdD-ChtBD but not PkdD exhibited strong binding to alpha-chitin and beta-chitin. Western and Northern analyses demonstrated that aprIV was induced in the presence of N-acetylglucosamine, N acetylchitobiose, or chitin. Native AprIV was purified to homogeneity from Alteromonas sp. strain O-7 and characterized. The molecular mass of mature AprIV was estimated to be 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimum pH and temperature of AprIV were pH 11.5 and 35 degrees C, respectively, and even at 10 degrees C the enzyme showed 25% of the maximum activity. Pretreatment of native chitin with AprIV significantly promoted chitinase activity. PMID- 11889093 TI - Strains of Escherichia coli O157:H7 differ primarily by insertions or deletions, not single-nucleotide polymorphisms. AB - Escherichia coli O157:H7 (O157) strains demonstrate varied pulsed-field gel electrophoresis patterns following XbaI digestion, which enable epidemiological surveillance of this important human pathogen. The genetic events underlying PFGE differences between strains, however, are not defined. We investigated the mechanisms for strain variation in O157 by recovering and examining nucleotide sequences flanking each of the XbaI restriction enzyme sites in the genome. Our analysis demonstrated that differences between O157 strains were due to discrete insertions or deletions that contained the XbaI sites polymorphic between strains rather than single-nucleotide polymorphisms in the XbaI sites themselves. These insertions and deletions were found to be uniquely localized within the regions of the genome that are specific to O157 compared to E. coli K-12 (O islands), suggesting that strain-to-strain variation occurs in these O islands. These results may be utilized to devise novel strain-typing tools for this pathogen. PMID- 11889094 TI - Identification of the cAD1 sex pheromone precursor in Enterococcus faecalis. AB - The Enterococcus faecalis virulence plasmid pAD1 encodes a mating response induced by exposure to an octapeptide sex pheromone, cAD1, secreted by plasmid free enterococci. The determinant for the pheromone in E. faecalis FA2-2, designated cad, was found to encode a 309-amino-acid lipoprotein precursor with the last 8 residues of its 22-amino acid signal sequence representing the cAD1 moiety. The lipoprotein moiety contained two 77-amino-acid repeats (70% identity) separated by 45 residues. The nonisogenic E. faecalis strain V583 determinant encodes a homologous precursor protein, but it differs at two amino acid positions, both of which are located within the pheromone peptide moiety (positions 2 and 8). Construction of a variant of strain FA2-2 containing the differences present in V583 resulted in cells that did not produce detectable cAD1. The mutant appeared normal under laboratory growth conditions, and while significantly reduced in recipient potential, when carrying pAD1 it exhibited a normal mating response. A mutant of FA2-2 with a truncated lipoprotein moiety appeared normal with respect to recipient potential and, when carrying plasmid DNA, donor potential. A gene encoding a protein designated Eep, believed to be a zinc metalloprotease, had been previously identified as required for pheromone biosynthesis and was believed to be involved in the processing of a pheromone precursor. Our new observation that the pAD1-encoded inhibitor peptide, iAD1, whose precursor is itself a signal sequence, is also dependent on Eep is consistent with the likelihood that such processing occurs at the amino terminus of the cAD1 moiety. PMID- 11889095 TI - Effects of Escherichia coli physiology on growth of phage T7 in vivo and in silico. AB - Phage development depends not only upon phage functions but also on the physiological state of the host, characterized by levels and activities of host cellular functions. We established Escherichia coli at different physiological states by continuous culture under different dilution rates and then measured its production of phage T7 during a single cycle of infection. We found that the intracellular eclipse time decreased and the rise rate increased as the growth rate of the host increased. To develop mechanistic insight, we extended a computer simulation for the growth of phage T7 to account for the physiology of its host. Literature data were used to establish mathematical correlations between host resources and the host growth rate; host resources included the amount of genomic DNA, pool sizes and elongation rates of RNA polymerases and ribosomes, pool sizes of amino acids and nucleoside triphosphates, and the cell volume. The in silico (simulated) dependence of the phage intracellular rise rate on the host growth rate gave quantitatively good agreement with our in vivo results, increasing fivefold for a 2.4-fold increase in host doublings per hour, and the simulated dependence of eclipse time on growth rate agreed qualitatively, deviating by a fixed delay. When the simulation was used to numerically uncouple host resources from the host growth rate, phage growth was found to be most sensitive to the host translation machinery, specifically, the level and elongation rate of the ribosomes. Finally, the simulation was used to follow how bottlenecks to phage growth shift in response to variations in host or phage functions. PMID- 11889096 TI - Isolation and characterization of cLV25, a Bacteroides fragilis chromosomal transfer factor resembling multiple Bacteroides sp. mobilizable transposons. AB - Horizontal DNA transfer contributes significantly to the dissemination of antibiotic resistance genes in Bacteroides fragilis. To further our understanding of DNA transfer in B. fragilis, we isolated and characterized a new transfer factor, cLV25. cLV25 was isolated from B. fragilis LV25 by its capture on the nonmobilizable Escherichia coli-Bacteroides shuttle vector pGAT400DeltaBglII. Similar to other Bacteroides sp. transfer factors, cLV25 was mobilized in E. coli by the conjugative plasmid R751. Using Tn1000 mutagenesis and deletion analysis of cLV25, two mobilization genes, bmgA and bmgB, were identified, whose predicted proteins have similarity to DNA relaxases and mobilization proteins, respectively. In particular, BmgA and BmgB were homologous to MocA and MocB, respectively, the two mobilization proteins of the B. fragilis mobilizable transposon Tn4399. A cis-acting origin of transfer (oriT) was localized to a 353 bp region that included nearly all of the intergenic region between bmgB and orf22 and overlapped with the 3' end of orf22. This oriT contained a putative nic site sequence but showed no significant similarity to the oriT regions of other transfer factors, including Tn4399. Despite the lack of sequence similarity between the oriTs of cLV25 and Tn4399, a mutation in the cLV25 putative DNA relaxase, bmgA, was partially complemented by Tn4399. In addition to the functional cross-reaction with Tn4399, a second distinguishing feature of cLV25 is that predicted proteins have similarity to proteins encoded not only by Tn4399 but by several Bacteroides sp. transfer factors, including NBU1, NBU2, CTnDOT, Tn4555, and Tn5520. PMID- 11889097 TI - Metabolic signals that lead to control of CBB gene expression in Rhodobacter capsulatus. AB - Various mutant strains were used to examine the regulation and metabolic control of the Calvin-Benson-Bassham (CBB) reductive pentose phosphate pathway in Rhodobacter capsulatus. Previously, a ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO)-deficient strain (strain SBI/II) was found to show enhanced levels of cbb(I) and cbb(II) promoter activities during photoheterotrophic growth in the presence of dimethyl sulfoxide. With this strain as the starting point, additional mutations were made in genes encoding phosphoribulokinase and transketolase and in the gene encoding the LysR-type transcriptional activator, CbbR(II). These strains revealed that a product generated by phosphoribulokinase was involved in control of CbbR-mediated cbb gene expression in SBI/II. Additionally, heterologous expression experiments indicated that Rhodobacter sphaeroides CbbR responded to the same metabolic signal in R. capsulatus SBI/II and mutant strain backgrounds. PMID- 11889099 TI - Membrane topology of the Streptococcus pneumoniae FtsW division protein. AB - The topology of FtsW from Streptococcus pneumoniae, an essential membrane protein involved in bacterial cell division, was predicted by computational methods and probed by the alkaline phosphatase fusion and cysteine accessibility techniques. Consistent results were obtained for the seven N-terminal membrane-spanning segments. However, the results from alkaline phosphatase fusions did not confirm the hydropathy analysis of the C-terminal part of FtsW, whereas the accessibility of introduced cysteine residues was in agreement with the theoretical prediction. Based on the combined results, we propose the first topological model of FtsW, featuring 10 membrane-spanning segments, a large extracytoplasmic loop, and both N and C termini located in the cytoplasm. PMID- 11889098 TI - Two distinct alcohol dehydrogenases participate in butane metabolism by Pseudomonas butanovora. AB - The involvement of two primary alcohol dehydrogenases, BDH and BOH, in butane utilization in Pseudomonas butanovora (ATCC 43655) was demonstrated. The genes coding for BOH and BDH were isolated and characterized. The deduced amino acid sequence of BOH suggests a 67-kDa alcohol dehydrogenase containing pyrroloquinoline quinone (PQQ) as cofactor and in the periplasm (29-residue leader sequence). The deduced amino acid sequence of BDH is consistent with a 70.9-kDa, soluble, periplasmic (37-residue leader sequence) alcohol dehydrogenase containing PQQ and heme c as cofactors. BOH and BDH mRNAs were induced whenever the cell's 1-butanol oxidation activity was induced. When induced with butane, the gene for BOH was expressed earlier than the gene for BDH. Insertional disruption of bdh or boh affected adversely, but did not eliminate, butane utilization by P. butanovora. The P. butanovora mutant with both genes boh and bdh inactivated was unable to grow on butane or 1-butanol. These cells, when grown in citrate and incubated in butane, developed butane oxidation capability and accumulated 1-butanol. The enzyme activity of BOH was characterized in cell extracts of the P. butanovora strain with bdh disrupted. Unlike BDH, BOH oxidized 2-butanol. The results support the involvement of two distinct NAD(+) independent, PQQ-containing alcohol dehydrogenases, BOH (a quinoprotein) and BDH (a quinohemoprotein), in the butane oxidation pathway of P. butanovora. PMID- 11889100 TI - Identification and characterization of CAMP cohemolysin as a potential virulence factor of Riemerella anatipestifer. AB - Riemerella anatipestifer is responsible for exudative septicemia in ducks. The genetic determinant of the CAMP cohemolysin, cam, from a strain of R. anatipestifer was cloned and expressed in Escherichia coli. Chromosomal DNA from serotype 19 strain 30/90 was used to construct a gene library in pBluescript II SK(-) vector in E. coli XL-1-Blue strain. The clones containing recombinant plasmids were screened for the CAMP reaction with Staphylococcus aureus. Those that showed cohemolysis were chosen for further analysis by sequencing. One of these clones, JFRA8, was subcloned to identify the smallest possible DNA fragment containing the CAMP cohemolysin determinant, which was located on a 3,566-bp BamHI-BstXI fragment which specified a 1,026-bp open reading frame. Clones containing recombinant plasmids carrying cam obtained by PCR cloning into E. coli M15 strain secreted an active CAMP cohemolysin. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analyses confirmed that the recombinant strain expressed a protein with a molecular mass of 37 kDa and that strains from serotypes 1, 2, 3, 5, 6, and 19 expressed the cohemolysin. The deduced amino acid sequence showed high homology to those of O-sialoglycoprotein endopeptidases. Hydrolysis of radioiodinated glycophorin A confirmed that Cam is a sialoglycoprotease. PMID- 11889101 TI - Selenium is mobilized in vivo from free selenocysteine and is incorporated specifically into formate dehydrogenase H and tRNA nucleosides. AB - Selenophosphate synthetase (SPS), the selD gene product from Escherichia coli, catalyzes the biosynthesis of monoselenophosphate, AMP, and orthophosphate in a 1:1:1 ratio from selenide and ATP. It was recently demonstrated that selenium delivered from selenocysteine by an E. coli NifS-like protein could replace free selenide in the in vitro SPS assay for selenophosphate formation (G. M. Lacourciere, H. Mihara, T. Kurihara, N. Esaki, and T. C. Stadtman, J. Biol. Chem. 275:23769-23773, 2000). During growth of E. coli in the presence of 0.1 microM (75)SeO(3)(2-) and increasing amounts of L-selenocysteine, a concomitant decrease in (75)Se incorporation into formate dehydrogenase H and nucleosides of bulk tRNA was observed. This is consistent with the mobilization of selenium from L selenocysteine in vivo and its use in selenophosphate formation. The ability of E. coli to utilize selenocysteine as a selenium source for selenophosphate biosynthesis in vivo supports the participation of the NifS-like proteins in selenium metabolism. PMID- 11889102 TI - Chemiosmotic energy conservation with Na(+) as the coupling ion during hydrogen dependent caffeate reduction by Acetobacterium woodii. AB - Cell suspensions of Acetobacterium woodii prepared from cultures grown on fructose plus caffeate catalyzed caffeate reduction with electrons derived from molecular hydrogen. Hydrogen-dependent caffeate reduction was strictly Na(+) dependent with a K(m) for Na(+) of 0.38 mM; Li(+) could substitute for Na(+). The sodium ionophore ETH2120, but not protonophores, stimulated hydrogen-dependent caffeate reduction by 280%, indicating that caffeate reduction is coupled to the buildup of a membrane potential generated by primary Na(+) extrusion. Caffeate reduction was coupled to the synthesis of ATP, and again, ATP synthesis coupled to hydrogen-dependent caffeate reduction was strictly Na(+) dependent and abolished by ETH2120, but not by protonophores, indicating the involvement of a transmembrane Na(+) gradient in ATP synthesis. The ATPase inhibitor N,N' dicyclohexylcarbodiimide (DCCD) abolished ATP synthesis, and at the same time, hydrogen-dependent caffeate reduction was inhibited. This inhibition could be relieved by ETH2120. These experiments are fully compatible with a chemiosmotic mechanism of ATP synthesis with Na(+) as the coupling ion during hydrogen dependent caffeate reduction by A. woodii. PMID- 11889103 TI - The pyrimidine nucleotide reductase step in riboflavin and F(420) biosynthesis in archaea proceeds by the eukaryotic route to riboflavin. AB - The Methanococcus jannaschii gene MJ0671 was cloned and overexpressed in Escherichia coli, and its gene product was tested for its ability to catalyze the pyridine nucleotide-dependent reduction of either 2,5-diamino-6-ribosylamino 4(3H)-pyrimidinone 5'-phosphate (compound 3) to 2,5-diamino-6-ribitylamino-4(3H) pyrimidinone 5'-phosphate (compound 4) or 5-amino-6-ribosylamino-2,4(1H,3H) pyrimidinedione 5'-phosphate (compound 7) to 5-amino-6-ribitylamino-2,4(1H,3H) pyrimidinedione 5'-phosphate (compound 5). Only compound 3 was found to serve as a substrate for the enzyme. NADPH and NADH functioned equally well as the reductants. This specificity for the reduction of compound 3 was also confirmed by using cell extracts of M. jannaschii and Methanosarcina thermophila. Thus, this step in riboflavin biosynthesis in these archaea is the same as that found in yeasts. The absence of the other genes in the biosynthesis of riboflavin in Archaea is discussed. PMID- 11889104 TI - Physical map of the linear chromosome of Streptomyces hygroscopicus 10-22 deduced by analysis of overlapping large chromosomal deletions. AB - The chromosomal DNA of Streptomyces hygroscopicus 10-22, a derivative of strain 5102-6, was digested with several restriction endonucleases and analyzed by pulsed-field gel electrophoresis (PFGE). Digestions with AseI gave 11 fragments with a total length of ca. 7.36 Mb. The AseI sites were mapped by analysis of overlapping chromosomal deletions in different mutants and confirmed by Southern hybridizations using partially digested genome fragments and linking cosmids as probes. PFGE analysis of DNA with and without proteinase K treatment, together with the hybridization results, suggested a linear organization with terminal proteins and large terminal inverted repeats. Some deletion mutants had circular chromosomes. PMID- 11889105 TI - Control of butanol formation in Clostridium acetobutylicum by transcriptional activation. AB - The sol operon of Clostridium acetobutylicum is the essential transcription unit for formation of the solvents butanol and acetone. The recent proposal that transcriptional regulation of this operon is controlled by the repressor Orf5/SolR (R. V. Nair, E. M. Green, D. E. Watson, G. N. Bennett, and E. T. Papoutsakis, J. Bacteriol. 181:319-330, 1999) was found to be incorrect. Instead, regulation depends on activation, most probably by the multivalent transcription factor Spo0A. The operon is transcribed from a single promoter. A second signal identified in primer extension studies results from mRNA processing and can be observed only in the natural host, not in a heterologous host. The first structural gene in the operon (adhE, encoding a bifunctional butyraldehyde/butanol dehydrogenase) is translated into two different proteins, the mature AdhE enzyme and the separate butanol dehydrogenase domain. The promoter of the sol operon is preceded by three imperfect repeats and a putative Spo0A-binding motif, which partially overlaps with repeat 3 (R3). Reporter gene analysis performed with the lacZ gene of Thermoanaerobacterium thermosulfurigenes and targeted mutations of the regulatory region revealed that the putative Spo0A binding motif, R3, and R1 are essential for control. The data obtained also indicate that an additional activator protein is involved. PMID- 11889106 TI - Complete genomic sequence of SfV, a serotype-converting temperate bacteriophage of Shigella flexneri. AB - Bacteriophage SfV is a temperate serotype-converting phage of Shigella flexneri. SfV encodes the factors involved in type V O-antigen modification, and the serotype conversion and integration-excision modules of the phage have been isolated and characterized. We now report on the complete sequence of the SfV genome (37,074 bp). A total of 53 open reading frames were predicted from the nucleotide sequence, and analysis of the corresponding proteins was used to construct a functional map. The general organization of the genes in the SfV genome is similar to that of bacteriophage lambda, and numerous features of the sequence are described. The superinfection immunity system of SfV includes a lambda-like repression system and a P4-like transcription termination mechanism. Sequence analysis also suggests that SfV encodes multiple DNA methylases, and experiments confirmed that orf-41 encodes a Dam methylase. Studies conducted to determine if the phage-encoded methylase confers host DNA methylation showed that the two S. flexneri strains analyzed encode their own Dam methylase. Restriction mapping and sequence analysis revealed that the phage genome has cos sites at the termini. The tail assembly and structural genes of SfV show homology to those of phage Mu and Mu-like prophages in the genome of Escherichia coli O157:H7 and Haemophilus influenzae. Significant homology (30% of the genome in total) between sections of the early, regulatory, and structural regions of the SfV genome and the e14 and KpLE1 prophages in the E. coli K-12 genome were noted, suggesting that these three phages have common evolutionary origins. PMID- 11889107 TI - Differential DNA binding of transcriptional regulator PcaU from Acinetobacter sp. strain ADP1. AB - Transcriptional regulator PcaU from Acinetobacter sp. strain ADP1 governs expression of genes for protocatechuate degradation (pca genes) as a repressor or an activator depending on the levels of the inducer protocatechuate and of its own gene. PcaU is a member of the IclR protein family. Here the DNA binding properties of the purified protein are described in terms of the location of the binding sites and the affinity to these sites. Native PcaU was purified after overexpression of the pcaU gene in Escherichia coli. It is a dimer in solution. The binding site in the pcaU-pcaI intergenic region is located between the two divergent promoters covering 45 bp, which includes three perfect 10-bp repetitions. A PcaU binding site downstream of pcaU is covered by PcaU across two palindromic sequence repetitions. The affinity of PcaU for the intergenic binding sites is 50-fold higher (dissociation constant [K(d)], 0.16 nM) than the affinity for the site downstream of pcaU (K(d), 8 nM). The binding of PcaU was tested after modifications of the intergenic binding site. Removal of any external sequence repetition still allowed for specific binding of PcaU, but the affinity was significantly reduced, suggesting an important role for all three sequence repetitions in gene expression. The involvement of DNA bending in the regulatory process is suggested by the observed strong intrinsic curvature displayed by the pcaU-pcaI intergenic DNA. PMID- 11889108 TI - Analysis of the Bacillus subtilis spoIIIJ gene and its Paralogue gene, yqjG. AB - The Bacillus subtilis spoIIIJ gene, which has been proven to be vegetatively expressed, has also been implicated as a sporulation gene. Recent genome sequencing information in many organisms reveals that spoIIIJ and its paralogous gene, yqjG, are conserved from prokaryotes to humans. A homologue of SpoIIIJ/YqjG, the Escherichia coli YidC is involved in the insertion of membrane proteins into the lipid bilayer. On the basis of this similarity, it was proposed that the two homologues act as translocase for the membrane proteins. We studied the requirements for spoIIIJ and yqjG during vegetative growth and sporulation. In rich media, the growth of spoIIIJ and yqjG single mutants were the same as that of the wild type, whereas spoIIIJ yqjG double inactivation was lethal, indicating that together these B. subtilis translocase homologues play an important role in maintaining the viability of the cell. This result also suggests that SpoIIIJ and YqjG probably control significantly overlapping functions during vegetative growth. spoIIIJ mutations have already been established to block sporulation at stage III. In contrast, disruption of yqjG did not interfere with sporulation. We further show that high level expression of spoIIIJ during vegetative phase is dispensable for spore formation, but the sporulation-specific expression of spoIIIJ is necessary for efficient sporulation even at the basal level. Using green fluorescent protein reporter to monitor SpoIIIJ and YqjG localization, we found that the proteins localize at the cell membrane in vegetative cells and at the polar and engulfment septa in sporulating cells. This localization of SpoIIIJ at the sporulation-specific septa may be important for the role of spoIIIJ during sporulation. PMID- 11889109 TI - Genome sequence and analysis of the oral bacterium Fusobacterium nucleatum strain ATCC 25586. AB - We present a complete DNA sequence and metabolic analysis of the dominant oral bacterium Fusobacterium nucleatum. Although not considered a major dental pathogen on its own, this anaerobe facilitates the aggregation and establishment of several other species including the dental pathogens Porphyromonas gingivalis and Bacteroides forsythus. The F. nucleatum strain ATCC 25586 genome was assembled from shotgun sequences and analyzed using the ERGO bioinformatics suite (http://www.integratedgenomics.com). The genome contains 2.17 Mb encoding 2,067 open reading frames, organized on a single circular chromosome with 27% GC content. Despite its taxonomic position among the gram-negative bacteria, several features of its core metabolism are similar to that of gram-positive Clostridium spp., Enterococcus spp., and Lactococcus spp. The genome analysis has revealed several key aspects of the pathways of organic acid, amino acid, carbohydrate, and lipid metabolism. Nine very-high-molecular-weight outer membrane proteins are predicted from the sequence, none of which has been reported in the literature. More than 137 transporters for the uptake of a variety of substrates such as peptides, sugars, metal ions, and cofactors have been identified. Biosynthetic pathways exist for only three amino acids: glutamate, aspartate, and asparagine. The remaining amino acids are imported as such or as di- or oligopeptides that are subsequently degraded in the cytoplasm. A principal source of energy appears to be the fermentation of glutamate to butyrate. Additionally, desulfuration of cysteine and methionine yields ammonia, H(2)S, methyl mercaptan, and butyrate, which are capable of arresting fibroblast growth, thus preventing wound healing and aiding penetration of the gingival epithelium. The metabolic capabilities of F. nucleatum revealed by its genome are therefore consistent with its specialized niche in the mouth. PMID- 11889111 TI - Filamentous phage active on the gram-positive bacterium Propionibacterium freudenreichii. AB - We present the first description of a single-stranded DNA filamentous phage able to replicate in a gram-positive bacterium. Phage B5 infects Propionibacterium freudenreichii and has a genome consisting of 5,806 bases coding for 10 putative open reading frames. The organization of the genome is very similar to the organization of the genomes of filamentous phages active on gram-negative bacteria. The putative coat protein exhibits homology with the coat proteins of phages PH75 and Pf3 active on Thermus thermophilus and Pseudomonas aeruginosa, respectively. B5 is, therefore, evolutionarily related to the filamentous phages active on gram-negative bacteria. PMID- 11889110 TI - Involvement of a cytochrome P450 monooxygenase in thaxtomin A biosynthesis by Streptomyces acidiscabies. AB - The biosynthesis of the thaxtomin cyclic dipeptide phytotoxins proceeds nonribosomally via the thiotemplate mechanism. Acyladenylation, thioesterification, N-methylation, and cyclization of two amino acid substrates are catalyzed by the txtAB-encoded thaxtomin synthetase. Nucleotide sequence analysis of the region 3' of txtAB in Streptomyces acidiscabies 84.104 identified an open reading frame (ORF) encoding a homolog of the P450 monooxygenase gene family. It was proposed that thaxtomin A phenylalanyl hydroxylation was catalyzed by the monooxygenase homolog. The ORF was mutated in S. acidiscabies 84.104 by using an integrative gene disruption construct, and culture filtrate extracts of the mutant were assayed for the presence of dehydroxy derivatives of thaxtomin A. Reversed-phase high-performance liquid chromatography (HPLC) and HPLC-mass spectrometry indicated that the major component in culture filtrate extracts of the mutant was less polar and smaller than thaxtomin A. Comparisons of electrospray mass spectra as well as (1)H- and (13)C-nuclear magnetic resonance spectra of the purified compound with those previously reported for thaxtomins confirmed the structure of the compound as 12,15-N-dimethylcyclo-(L-4 nitrotryptophyl-L-phenylalanyl), the didehydroxy analog of thaxtomin A. The ORF, designated txtC, was cloned and the recombinant six-His-tagged fusion protein produced in Escherichia coli and purified from cell extracts. TxtC produced in E. coli exhibited spectral properties similar to those of cytochrome P450-type hemoproteins that have undergone conversion to the catalytically inactive P420 form. Based on these properties and the high similarity of TxtC to other well characterized P450 enzymes, we conclude that txtC encodes a cytochrome P450-type monooxygenase required for postcyclization hydroxylation of the cyclic dipeptide. PMID- 11889112 TI - Perfringolysin O expression in Clostridium perfringens is independent of the upstream pfoR gene. AB - The pathogenesis of Clostridium perfringens-mediated gas gangrene or clostridial myonecrosis involves the extracellular toxins alpha-toxin and perfringolysin O. Previous studies (T. Shimizu, A. Okabe, J. Minami, and H. Hayashi, Infect. Immun. 59:137-142, 1991) carried out with Escherichia coli suggested that the perfringolysin O structural gene, pfoA, was positively regulated by the product of the upstream pfoR gene. In an attempt to confirm this hypothesis in C. perfringens, a pfoR-pfoA deletion mutant was complemented with isogenic pfoA(+) shuttle plasmids that varied only in their ability to encode an intact pfoR gene. No difference in the ability to produce perfringolysin O was observed for C. perfringens strains carrying these plasmids. In addition, chromosomal pfoR mutants were constructed by homologous recombination in C. perfringens. Again no difference in perfringolysin O activity was observed. Since it was not possible to alter perfringolysin O expression by mutation of pfoR, it was concluded that the pfoR gene product is unlikely to have a role in the regulation of pfoA expression in C. perfringens. PMID- 11889113 TI - Regulation of purine hydroxylase and xanthine dehydrogenase from Clostridium purinolyticum in response to purines, selenium, and molybdenum. AB - The discovery that two distinct enzyme catalysts, purine hydroxylase (PH) and xanthine dehydrogenase (XDH), are required for the overall conversion of hypoxanthine to uric acid by Clostridium purinolyticum was unexpected. In this reaction sequence, hypoxanthine is hydroxylated to xanthine by PH and then xanthine is hydroxylated to uric acid by XDH. PH and XDH, which contain a labile selenium cofactor in addition to a molybdenum cofactor, flavin adenine dinucleotide, and FeS centers, were purified and partially characterized as reported previously. In the present study, the activities of these two enzymes were measured in cells grown in media containing various concentrations of selenite, molybdate, and various purine substrates. The levels of PH protein in extracts were determined by immunoblot assay. The amount of PH protein, as well as the specific activities of PH and XDH, increased when either selenite or molybdate was added to the culture medium. PH levels were highest in the cells cultured in the presence of either adenine or purine. XDH activity increased dramatically in cells grown with either xanthine or uric acid. The apparent increases in protein levels and activities of PH and XDH in response to selenium, molybdenum, and purine substrates demonstrate that these enzymes are tightly regulated in response to these nutrients. PMID- 11889114 TI - Differential effects of mutations in tonB1 on intrinsic multidrug resistance and iron acquisition in Pseudomonas aeruginosa. AB - Loss of tonB1 adversely affects iron acquisition and intrinsic multidrug resistance in Pseudomonas aeruginosa. Several mutations in tonB1 compromised the protein's contribution to both processes, although TonB1 derivatives altered in residues C35, Q268, R287, Q292, R300, and R304 were compromised vis-a-vis their contribution to drug resistance only. PMID- 11889115 TI - Evidence for lateral transfer of the Suilysin gene region of Streptococcus suis. AB - Suilysin is a cholesterol-binding cytolysin encoded by sly in Streptococcus suis. DNA sequence determination of the sly locus in a strain lacking sly revealed the presence of another gene, designated orf102, in the place of sly. No transposable element or long-repeat sequence was found in the close vicinity. Except for six strains whose corresponding loci have been rearranged, all of the remaining 62 strains examined had either sly or orf102 at the same locus and their flanking regions were conserved. The genetic organizations having either sly or orf102 were found in the strains whose 16S rRNA sequences were identical. These results suggest that S. suis acquired sly or orf102 from a foreign source and that these genes subsequently spread among S. suis strains by homologous recombination. PMID- 11889117 TI - Phosphorylation of p27Kip1 on serine 10 is required for its binding to CRM1 and nuclear export. AB - Phosphorylation of the cyclin-dependent kinase inhibitor p27(Kip1) has been thought to regulate its stability. Ser(10) is the major phosphorylation site of p27(Kip1), and phosphorylation of this residue affects protein stability. Phosphorylation of p27(Kip1) on Ser(10) has now been shown to be required for the binding of CRM1, a carrier protein for nuclear export. The p27(Kip1) protein was translocated from the nucleus to the cytoplasm at the G(0)-G(1) transition of the cell cycle, and this export was inhibited by leptomycin B, a specific inhibitor of CRM1-dependent nuclear export. The nuclear export and subsequent degradation of p27(Kip1) at the G(0)-G(1) transition were observed in cells lacking Skp2, the F-box protein component of an SCF ubiquitin ligase complex, indicating that these early events are independent of Skp2-mediated proteolysis. Substitution of Ser(10) with Ala (S10A) markedly reduced the extent of p27(Kip1) export, whereas substitution of Ser(10) with Asp (S10D) or Glu (S10E) promoted export. Co immunoprecipitation analysis showed that CRM1 preferentially interacted with S10D and S10E but not with S10A, suggesting that the phosphorylation of p27(Kip1) on Ser(10) is required for its binding to CRM1 and for its subsequent nuclear export. PMID- 11889116 TI - Conserved serine/threonine kinase encoded by CBK1 regulates expression of several hypha-associated transcripts and genes encoding cell wall proteins in Candida albicans. AB - The opportunistic fungal pathogen, Candida albicans, is reported to have several potential virulence factors. A potentially significant factor is the ability to undergo morphological transition from yeast to hypha. This alteration of form is accompanied by many changes within the cell, including alterations in gene expression and cell wall composition. We have isolated a gene that encodes a highly conserved serine/threonine kinase that appears to be involved in the regulation of proteins associated with the cell wall. We have assigned the designation CBK1 (cell wall biosynthesis kinase 1) to this gene. Mutants lacking CBK1 form large aggregates of round cells under all growth conditions and lack the ability to undergo morphological differentiation. Additionally, these mutants show an altered pattern of expression of several transcripts encoding proteins associated with the cell wall. The results suggest that the kinase encoded by CBK1 plays a general role in the maintenance and alteration of the cell wall of C. albicans in all morphologies. PMID- 11889118 TI - Cooperative action of Escherichia coli ClpB protein and DnaK chaperone in the activation of a replication initiation protein. AB - The Escherichia coli molecular chaperone protein ClpB is a member of the highly conserved Hsp100/Clp protein family. Previous studies have shown that the ClpB protein is needed for bacterial thermotolerance. Purified ClpB protein has been shown to reactivate chemically and heat-denatured proteins. In this work we demonstrate that the combined action of ClpB and the DnaK, DnaJ, and GrpE chaperones leads to the activation of DNA replication of the broad-host-range plasmid RK2. In contrast, ClpB is not needed for the activation of the oriC dependent replication of E. coli. Using purified protein components we show that the ClpB/DnaK/DnaJ/GrpE synergistic action activates the plasmid RK2 replication initiation protein TrfA by converting inactive dimers to an active monomer form. In contrast, Hsp78/Ssc1/Mdj1/Mge1, the corresponding protein system from yeast mitochondria, cannot activate the TrfA replication protein. Our results demonstrate for the first time that the ClpB/DnaK/DnaJ/GrpE system is involved in protein monomerization and in the activation of a DNA replication factor. PMID- 11889120 TI - Inhibition of collagen alpha 1(I) expression by the 5' stem-loop as a molecular decoy. AB - Collagen alpha1(I) mRNA is posttranscriptionally regulated in hepatic stellate cells (HSCs). Binding of protein factors to the evolutionary conserved stem-loop in the 5'-untranslated region (5' stem-loop) is required for a high level of expression in activated HSCs. The 5' stem-loop is also found in alpha2(I) and alpha1(III) mRNAs. Titration of the 5' stem-loop binding factors by a stably expressed RNA containing the 5' stem-loop (molecular decoy) may decrease the expression of these collagen mRNAs. We designed a 108-nt RNA that is transcribed from the optimized mouse U7 small nuclear RNA gene and contains the 5' stem-loop (p74WT decoy). This decoy accumulates in the nucleus and in the cytoplasm. When expressed in NIH 3T3 fibroblasts, the p74WT decoy decreased collagen alpha1(I) mRNA level by 60% and decreased collagen type I secreted into the cellular medium by 50%. We also expressed this decoy in quiescent rat HSCs by adenoviral gene transfer. Quiescent HSCs undergo activation in culture, resulting in a 60-70-fold increase in collagen alpha1(I) mRNA. The decoy decreases collagen alpha1(I) mRNA expression by 50-60% during activation of HSCs. It also decreases collagen alpha2(I) mRNA expression and collagen alpha1(III) mRNA expression. The cellular levels of collagen alpha1(I) propeptide and of disulfide-bonded collagen type I trimer are reduced by 70%. However, the p74WT decoy did not decrease alpha smooth muscle actin protein or the mRNA levels of glyceraldehyde-3-phosphate dehydrogenase and interleukin-6. The p74WT decoy was also introduced into activated human HSCs. In these cells, the decoy decreased collagen alpha1(I) propeptide and disulfide-bonded collagen trimer by 50-60%. These results indicate that the 5' stem-loop specifically regulates fibrillar collagen synthesis and represents a novel target for antifibrotic therapy. The molecular decoys provide a generalized method of assessing the functional significance of blocking the interactions of mRNA and proteins. PMID- 11889119 TI - Csx/Nkx2-5 is required for homeostasis and survival of cardiac myocytes in the adult heart. AB - Csx/Nkx2-5, which is essential for cardiac development of the embryo, is abundantly expressed in the adult heart. We here examined the role of Csx/Nkx2-5 in the adult heart using two kinds of transgenic mice. Transgenic mice that overexpress a dominant negative mutant of Csx/Nkx2-5 (DN-TG mice) showed degeneration of cardiac myocytes and impairment of cardiac function. Doxorubicin induced more marked cardiac dysfunction in DN-TG mice and less in transgenic mice that overexpress wild type Csx/Nkx2-5 (WT-TG mice) compared with non-transgenic mice. Doxorubicin induced cardiomyocyte apoptosis, and the number of apoptotic cardiomyocytes was high in the order of DN-TG mice, non-transgenic mice, and WT TG mice. Overexpression of the dominant negative mutant of Csx/Nkx2-5 induced apoptosis in cultured cardiomyocytes, while expression of wild type Csx/Nkx2-5 protected cardiomyocytes from doxorubicin-induced apoptotic death. These results suggest that Csx/Nkx2-5 plays a critical role in maintaining highly differentiated cardiac phenotype and in protecting the heart from stresses including doxorubicin. PMID- 11889121 TI - Guanylin, uroguanylin, and heat-stable euterotoxin activate guanylate cyclase C and/or a pertussis toxin-sensitive G protein in human proximal tubule cells. AB - Membrane guanylate cyclase C (GC-C) is the receptor for guanylin, uroguanylin, and heat-stable enterotoxin (STa) in the intestine. GC-C-deficient mice show resistance to STa in intestine but saluretic and diuretic effects of uroguanylin and STa are not disturbed. Here we describe the cellular effects of these peptides using immortalized human kidney epithelial (IHKE-1) cells with properties of the proximal tubule, analyzed with the slow-whole-cell patch clamp technique. Uroguanylin (10 or 100 nm) either hyperpolarized or depolarized membrane voltages (V(m)). Guanylin and STa (both 10 or 100 nm), as well as 8-Br cGMP (100 microm), depolarized V(m). All peptide effects were absent in the presence of 1 mm Ba(2+). Uroguanylin and guanylin changed V(m) pH dependently. Pertussis toxin (1 microg/ml, 24 h) inhibited hyperpolarizations caused by uroguanylin. Depolarizations caused by guanylin and uroguanylin were blocked by the tyrosine kinase inhibitor, genistein (10 microm). All three peptides increased cellular cGMP. mRNA for GC-C was detected in IHKE-1 cells and in isolated human proximal tubules. In IHKE-1 cells GC-C was also detected by immunostaining. These findings suggest that GC-C is probably the receptor for guanylin and STa. For uroguanylin two distinct signaling pathways exist in IHKE-1 cells, one involves GC-C and cGMP as second messenger, the other is cGMP independent and connected to a pertussis toxin-sensitive G protein. PMID- 11889122 TI - Microheterogeneity controls the rate of gelation of actin filament networks. AB - Rapid sol-gel transitions of the actin cytoskeleton are required for many key cellular processes, including cell spreading and cell locomotion. Actin monomers assemble into semiflexible polymers that rapidly intertwine into a network, a process that in vitro takes approximately 1 min for an actin concentration of 1 mg/ml. The same actin filament network, however, takes approximately 1 h to exhibit a steady-state elasticity. We hypothesize that the slow gelation of F actin is due to the slow establishment of a homogeneous meshwork. Using a novel method, time-resolved multiple particle tracking, which monitors the range of thermally excited displacements of microspheres imbedded in the network, we show that the increase in elasticity in a polymerizing solution of actin parallels the progressive decline of the network microheterogeneity. The rates of gelation and network homogenization slightly decrease with actin concentration and in the presence of the F-actin cross-linking proteins alpha-actinin and fascin, whereas the rate of actin polymerization increases dramatically with actin concentration. Our measurements show that the slow spatial homogenization of the actin filament network, not actin polymerization or the formation of polymer overlaps, is the rate-limiting step in the establishment of an elastic actin network and suggest that a new activity of F-actin binding proteins may be required for the rapid formation of a homogeneous stiff gel. PMID- 11889123 TI - Werner protein is a target of DNA-dependent protein kinase in vivo and in vitro, and its catalytic activities are regulated by phosphorylation. AB - Human Werner Syndrome is characterized by early onset of aging, elevated chromosomal instability, and a high incidence of cancer. Werner protein (WRN) is a member of the recQ gene family, but unlike other members of the recQ family, it contains a unique 3'-->5' exonuclease activity. We have reported previously that human Ku heterodimer interacts physically with WRN and functionally stimulates WRN exonuclease activity. Because Ku and DNA-PKcs, the catalytic subunit of DNA dependent protein kinase (DNA-PK), form a complex at DNA ends, we have now explored the possibility of functional modulation of WRN exonuclease activity by DNA-PK. We find that although DNA-PKcs alone does not affect the WRN exonuclease activity, the additional presence of Ku mediates a marked inhibition of it. The inhibition of WRN exonuclease by DNA-PKcs requires the kinase activity of DNA PKcs. WRN is a target for DNA-PKcs phosphorylation, and this phosphorylation requires the presence of Ku. We also find that treatment of recombinant WRN with a Ser/Thr phosphatase enhances WRN exonuclease and helicase activities and that WRN catalytic activity can be inhibited by rephosphorylation of WRN with DNA-PK. Thus, the level of phosphorylation of WRN appears to regulate its catalytic activities. WRN forms a complex, both in vitro and in vivo, with DNA-PKC. WRN is phosphorylated in vivo after treatment of cells with DNA-damaging agents in a pathway that requires DNA-PKcs. Thus, WRN protein is a target for DNA-PK phosphorylation in vitro and in vivo, and this phosphorylation may be a way of regulating its different catalytic activities, possibly in the repair of DNA dsb. PMID- 11889124 TI - A quality control pathway that down-regulates aberrant T-cell receptor (TCR) transcripts by a mechanism requiring UPF2 and translation. AB - Nonsense-mediated decay (NMD) is an RNA surveillance pathway that degrades mRNAs containing premature termination codons (PTC). T-cell receptor (TCR) and immunoglobulin (Ig) transcripts, which are encoded by genes that very frequently acquire PTCs during lymphoid ontogeny, are down-regulated much more dramatically in response to PTCs than are other known transcripts. Another feature unique to TCR, Ig, and a subset of other mRNAs is that they are down-regulated in response to nonsense codons in the nuclear fraction of cells. This is paradoxical, as the only well recognized entity that recognizes nonsense codons is the cytoplasmic translation apparatus. Therefore, we investigated whether translation is responsible for this nuclear-associated mechanism. We found that the down regulation of TCR-beta transcripts in response to nonsense codons requires several features of translation, including an initiator ATG and the ability to scan. We also found that optimal down-regulation depends on a Kozak consensus sequence surrounding the initiator ATG and that it can be initiated by an internal ribosome entry site, neither of which has been demonstrated before for any other PTC-bearing mRNA. At least a portion of this down-regulatory response is mediated by the NMD pathway as antisense hUPF2 transcripts increased the levels of PTC-bearing TCR-beta transcripts in the nuclear fraction of cells. We conclude that a hUPF2-dependent RNA surveillance pathway with translation-like features operating in the nuclear fraction of cells prevents the expression of potentially deleterious truncated proteins encoded by non-productively rearranged TCR genes. PMID- 11889125 TI - Control of myeloid-specific integrin alpha Mbeta 2 (CD11b/CD18) expression by cytokines is regulated by Stat3-dependent activation of PU.1. AB - Granulocyte colony-stimulating factor (G-CSF) plays an essential role in regulating multiple aspects of hematopoiesis. To elucidate the role of G-CSF in controlling hematopoietic cell migration capabilities, we studied inducible expression of the myeloid-specific marker, integrin alpha(M)beta(2) (CD11b/CD18, Mac-1), in the myeloid cell line, 32D. We found that G-CSF stimulates the synthesis and cell surface expression of alpha(M) and beta(2) integrin subunits. Induction of both alpha(M) and beta(2) is dependent on Stat3, a major G-CSF responsive signaling protein. However, the kinetics of expression suggested the involvement of an intermediate protein regulated by Stat3. Our results demonstrate that Stat3 signaling stimulates the expression of PU.1, a critical regulator of myelopoiesis. Furthermore, we show that PU.1 is an essential intermediate for the inducible expression of alpha(M)beta(2) integrin. Thus, Stat3 promotes alpha(M)beta(2) integrin expression through its activation of PU.1. These findings indicate that G-CSF-dependent Stat3 signals stimulate the changes in cell adhesion and migration capabilities that occur during myeloid cell development. These data also demonstrate a link between Stat3 and PU.1, suggesting that Stat3 may play an instructive role in hematopoiesis. PMID- 11889126 TI - HIP1 and HIP12 display differential binding to F-actin, AP2, and clathrin. Identification of a novel interaction with clathrin light chain. AB - Huntingtin-interacting protein 1 (HIP1) and HIP12 are orthologues of Sla2p, a yeast protein with essential functions in endocytosis and regulation of the actin cytoskeleton. We now report that HIP1 and HIP12 are major components of the clathrin coat that interact but differ in their ability to bind clathrin and the clathrin adaptor AP2. HIP1 contains a clathrin-box and AP2 consensus-binding sites that display high affinity binding to the terminal domain of the clathrin heavy chain and the ear domain of the AP2 alpha subunit, respectively. These consensus sites are poorly conserved in HIP12 and correspondingly, HIP12 does not bind to AP2 nor does it demonstrate high affinity clathrin binding. Moreover, HIP12 co-sediments with F-actin in contrast to HIP1, which exhibits no interaction with actin in vitro. Despite these differences, both proteins efficiently stimulate clathrin assembly through their central helical domain. Interestingly, in both HIP1 and HIP12, this domain binds directly to the clathrin light chain. Our data suggest that HIP1 and HIP12 play related yet distinct functional roles in clathrin-mediated endocytosis. PMID- 11889127 TI - Error prone translesion synthesis past gamma-hydroxypropano deoxyguanosine, the primary acrolein-derived adduct in mammalian cells. AB - 8-Hydroxy-5,6,7,8-tetrahydropyrimido[1,2-a]purin- 10(3H)-one,3-(2'-deoxyriboside) (1,N(2)-gamma-hydroxypropano deoxyguanosine, gamma-HOPdG) is a major DNA adduct that forms as a result of exposure to acrolein, an environmental pollutant and a product of endogenous lipid peroxidation. gamma-HOPdG has been shown previously not to be a miscoding lesion when replicated in Escherichia coli. In contrast to those prokaryotic studies, in vivo replication and mutagenesis assays in COS-7 cells using single stranded DNA containing a specific gamma-HOPdG adduct, revealed that the gamma-HOPdG adduct was significantly mutagenic. Analyses revealed both transversion and transition types of mutations at an overall mutagenic frequency of 7.4 x 10(-2)/translesion synthesis. In vitro gamma-HOPdG strongly blocks DNA synthesis by two major polymerases, pol delta and pol epsilon. Replicative blockage of pol delta by gamma-HOPdG could be diminished by the addition of proliferating cell nuclear antigen, leading to highly mutagenic translesion bypass across this adduct. The differential functioning and processing capacities of the mammalian polymerases may be responsible for the higher mutation frequencies observed in this study when compared with the accurate and efficient nonmutagenic bypass observed in the bacterial system. PMID- 11889128 TI - Elastase-released epidermal growth factor recruits epidermal growth factor receptor and extracellular signal-regulated kinases to down-regulate tropoelastin mRNA in lung fibroblasts. AB - Elastase/anti-elastase imbalance is a hallmark of emphysema, a chronic obstructive pulmonary disease associated with the rupture and inefficient repair of interstitial elastin. We report that neutrophil elastase (NE) at low physiologic concentrations, ranging from 35 nm to 1 microm, invokes transient, peaking at 15 min, activation of extracellular signal-regulated kinases 1 and 2 (ERK) in elastogenic lung fibroblasts. ERK activation is preceded by the release of soluble 25-26-kDa forms of epidermal growth factor (EGF) and transactivation of EGF receptor (EGFR) in NE-exposed cells. The stimulatory effect of NE on ERK is abrogated in the presence of anti-EGF-neutralizing antibodies, EGFR tyrosine kinase inhibitor (AG1478), and ERK kinase inhibitor (PD98059), as well as abolished in both EGFR-desensitized and endocytosis-arrested fibroblasts. Nuclear accumulation of activated ERK is associated with transient, peaking at 30 min, induction of c-Fos and sustained, observed at 24-48 h, decrease of tropoelastin mRNA levels in NE-challenged cells. Pretreatment of fibroblasts with AG1478 or PD98059 abrogates the NE-initiated tropoelastin mRNA suppression. We conclude that proteolytically released EGF signals directly via EGFR and ERK to down regulate tropoelastin mRNA in NE-challenged lung fibroblasts. PMID- 11889130 TI - Manipulation of cholesterol levels in rod disk membranes by methyl-beta cyclodextrin: effects on receptor activation. AB - The effect of cholesterol on rod outer segment disk membrane structure and rhodopsin activation was investigated. Disk membranes with varying cholesterol concentrations were prepared using methyl-beta-cyclodextrin as a cholesterol donor or acceptor. Cholesterol exchange followed a simple equilibrium partitioning model with a partition coefficient of 5.2 +/- 0.8 in favor of the disk membrane. Reduced cholesterol in disk membranes resulted in a higher proportion of photolyzed rhodopsin being converted to the G protein-activating metarhodopsin II (MII) conformation, whereas enrichment of cholesterol reduced the extent of MII formation. Time-resolved fluorescence anisotropy measurements using 1,6-diphenyl-1,3,5-hexatriene showed that increasing cholesterol reduced membrane acyl chain packing free volume as characterized by the parameter f(v). The level of MII formed showed a positive linear correlation with f(v) over the range of 4 to 38 mol % cholesterol. In addition, the thermal stability of rhodopsin increased with mol % of cholesterol in disk membranes. No evidence was observed for the direct interaction of cholesterol with rhodopsin in either its agonist- or antagonist-bound form. These results indicate that cholesterol mediates the function of the G protein-coupled receptor, rhodopsin, by influencing membrane lipid properties, i.e. reducing acyl chain packing free volume, rather than interacting specifically with rhodopsin. PMID- 11889129 TI - Structural rearrangement of human lymphotactin, a C chemokine, under physiological solution conditions. AB - NMR spectra of human lymphotactin (hLtn), obtained under various solution conditions, have revealed that the protein undergoes a major conformational rearrangement dependent on temperature and salt concentration. At high salt (200 mm NaCl) and low temperature (10 degrees C), hLtn adopts a chemokine-like fold, which consists of a three-stranded antiparallel beta-sheet and a C-terminal alpha helix (Kuloglu, E. S., McCaslin, D. R., Kitabwalla, M., Pauza, C. D., Markley, J. L., and Volkman, B. F. (2001) Biochemistry 40, 12486-12496). We have used NMR spectroscopy, sedimentation equilibrium, and intrinsic fluorescence to monitor the reversible conformational change undergone by hLtn as a function of temperature and ionic strength. We have used two-, three- and four-dimensional NMR spectroscopy of isotopically enriched protein samples to determine structural properties of the conformational state stabilized at 45 degrees C and 0 mm NaCl. Patterns of NOEs and (1)H(alpha) and (13)C chemical shifts show that hLtn rearranges under these conditions to form a four-stranded, antiparallel beta sheet with a pattern of hydrogen bonding that is completely different from that of the chemokine fold stabilized at 10 degrees C and 200 mm NaCl. The C-terminal alpha-helix observed at 10 degrees C and 200 mm NaCl, which is conserved in other chemokines, is absent at 45 degrees C and no salt, and the last 38 residues of the protein are completely disordered, as indicated by heteronuclear (15)N-(1)H NOEs. Temperature dependence of the tryptophan fluorescence of hLtn in low and high salt confirmed that the chemokine conformation is stabilized by increased ionic strength. Sedimentation equilibrium analytical ultracentrifugation showed that hLtn at 40 degrees C in the presence of 100 mm NaCl exists mainly as a dimer. Under near physiological conditions of temperature, pH, and ionic strength, both the chemokine-like and non-chemokine-like conformations of hLtn are significantly populated. The functional relevance of this structural interconversion remains to be elucidated. PMID- 11889131 TI - Clustering induces redistribution of syndecan-4 core protein into raft membrane domains. AB - Syndecan-4 is a heparan sulfate-carrying core protein that has been directly implicated in fibroblast growth factor 2 (FGF2) signaling. Recent studies have suggested that many signaling proteins localize to the raft compartment of the plasma cell membrane. To establish whether syndecan-4 is present in the raft compartment, we have studied the distribution of the core protein and an Fc receptor (FcR)-syndecan-4 chimera prior to and following clustering with FGF2 or antibodies. Whereas unclustered syndecan-4 was present predominantly in the non raft membrane compartment, clustering induced extensive syndecan-4 redistribution to the rafts as demonstrated by the sucrose gradient centrifugation and life confocal microscopy. Although syndecan-4 and caveolin-1 moved in tandem, syndecan 4 was not present in caveolae, a major subset of raft compartments. We conclude that syndecan-4 clustering induces its redistribution to the non-caveolae raft compartment. This process may play an important role in syndecan-4-mediation of FGF2 signaling. PMID- 11889132 TI - Regulation of ALF gene expression in somatic and male germ line tissues involves partial and site-specific patterns of methylation. AB - ALF (TFIIAalpha/beta-like factor) is a germ cell-specific counterpart of the large (alpha/beta) subunit of general transcription factor TFIIA. Here we isolated homologous GC-rich promoters from the mouse and human ALF genes and used promoter deletion analysis to identify sequences active in COS-7 and 293 cells. Further, bisulfite sequence analysis of the mouse ALF promoter showed that all 21 CpG dinucleotides between -179 and +207 were partially methylated in five somatic tissues, brain, heart, liver, lung, and muscle, and in epididymal spermatozoa from adult mice. In contrast, DNA from prepubertal mouse testis and from purified spermatocytes were unmethylated except at C(+19)G and C(+170)G. We also found that ALF expression correlates with a strong promoter-proximal DNase I hypersensitive site present in nuclei from testis but not from liver. Finally we show that in vitro methylation of the ALF promoter inhibits activity and that 5 aza-2'-deoxycytidine treatment reactivates the endogenous ALF gene in a panel of seven different mouse and human somatic cell lines. Overall the results show that silencing in somatic cells is methylation-dependent and reversible and that a unique CpG-specific methylation pattern at the ALF promoter precedes expression in pachytene spermatocytes. This pattern is transient as remethylation of the ALF promoter in haploid germ cell DNA has occurred by the time spermatozoa are present in the epididymis. PMID- 11889133 TI - The expression of keratin k10 in the basal layer of the epidermis inhibits cell proliferation and prevents skin tumorigenesis. AB - Forced expression of K10, a keratin normally expressed in postmitotic, terminally differentiating epidermal keratinocytes, inhibits the progression of the cell cycle in cultured cells (Paramio, J. M., Casanova, M. Ll., Segrelles, C., Mittnacht, S., Lane, E. B., and Jorcano, J. L. (1999) Mol. Cell. Biol. 19, 3086 3094). This process requires a functional retinoblastoma (pRb) gene product and is mediated by K10-induced inhibition of Akt and PKCzeta, two signaling intermediates belonging to the phosphoinositide (PI) 3-kinase signal transduction pathway (Paramio, J. M., Segrelles, C., Ruiz, S., and Jorcano, J. L. (2001) Mol. Cell. Biol. 21, 7449-7459). Extending earlier in vitro studies to the in vivo situation, this work analyzes the alterations found in transgenic mice that ectopically express K10 in the proliferative basal cells of the epidermis. Increased expression of K10 led to a hypoplastic and hyperkeratotic epidermis due to a dramatic decrease in skin keratinocyte proliferation in association with the inhibition of Akt and PKCzeta activities. The inhibition of cell proliferation and Akt and PKCzeta activities was also observed although to a minor extent in low hK10-expressing mice. These animals displayed no overt epidermal phenotype nor overexpression of K10. In these non-phenotypic mice, ectopic K10 expression also resulted in decreased skin tumorigenesis. Collectively, these data demonstrate that keratin K10 in vivo functions include the control of epithelial proliferation in skin epidermis. PMID- 11889134 TI - Relaxed acyl chain specificity of Bordetella UDP-N-acetylglucosamine acyltransferases. AB - Lipid A (endotoxin) is a major structural component of Gram-negative outer membranes. It also serves as the hydrophobic anchor of lipopolysaccharide and is a potent activator of the innate immune response. Lipid A molecules from the genus Bordetella are reported to exhibit unusual structural asymmetry with respect to the acyl chains at the 3- and 3'-positions. These acyl chains are attached by UDP-N-acetylglucosamine acyltransferase (LpxA). To determine the origin of the acyl variability, the single lpxA ortholog present in each of the genomes of Bordetella bronchiseptica (lpxA(Br)), Bordetella parapertussis (lpxA(Pa)), and Bordetella pertussis (lpxA(Pe)) was cloned and expressed in Escherichia coli. In contrast to all LpxA proteins studied to date, LpxA(Br) and LpxA(Pe) display relaxed acyl chain length specificity in vitro, utilizing C(10)OH-ACP, C(12)OH-ACP, and C(14)OH-ACP at similar rates. Furthermore, hybrid lipid A molecules synthesized at 42 degrees C by an E. coli lpxA mutant complemented with lpxA(Pe) contain C(10)OH, C(12)OH, and C(14)OH at both the 3- and 3'-positions, as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In contrast, LpxA from B. parapertussis did not display relaxed specificity but was selective for C(10)OH-ACP. This study provides an enzymatic explanation for some of the unusual acyl chain variations found in Bordetella lipid A. PMID- 11889135 TI - GSTB1-1 from Proteus mirabilis: a snapshot of an enzyme in the evolutionary pathway from a redox enzyme to a conjugating enzyme. AB - The native form of the bacterial glutathione transferase B1-1 (EC ) is characterized by one glutathione (GSH) molecule covalently linked to Cys-10. This peculiar disulfide, only found in the Beta and Omega class glutathione S transferases (GSTs) but absent in all other GSTs, prompts questions about its role and how GSH can be activated and utilized in the reaction normally performed by GSTs. Stopped-flow and spectroscopic experiments suggest that, in the native enzyme (GSTB1-1ox), a second GSH molecule is present, albeit transiently, in the active site. This second GSH binds to the enzyme through a bimolecular interaction followed by a fast thiol-disulfide exchange with the covalently bound GSH. The apparent pK(a) of the non-covalently bound GSH is lowered from 9.0 to 6.4 +/- 0.2 in similar fashion to other GSTs. The reduced form of GSTB1-1 (GSTB1 1red) binds GSH 100-fold faster and also induces a more active deprotonation of the substrate with an apparent pK(a) of 5.2 +/- 0.1. Apparently, the absence of the mixed disulfide does not affect k(cat) and K(m) values in the GST conjugation activity, which is rate-limited by the chemical step both in GSTB1-1red and in GSTB1-1ox. However, GSTB1-1ox follows a steady-state random sequential mechanism whereas a rapid-equilibrium random sequential mechanism is adopted by GSTB1-1red. Remarkably, GSTB1-1ox and GSTB1-1red are equally able to catalyze a glutaredoxin like catalysis using cysteine S-sulfate and hydroxyethyl disulfide as substrates. Cys-10 is an essential residue in this redox activity, and its replacement by alanine abolishes this enzymatic activity completely. It appears that GSTB1-1 behaves like an "intermediate enzyme" between the thiol-disulfide oxidoreductase and the GST superfamilies. PMID- 11889136 TI - Unconventional activation mechanisms of MMP-26, a human matrix metalloproteinase with a unique PHCGXXD cysteine-switch motif. AB - ProMMP-26 has the unique Pro-His(81)-Cys-Gly-Xaa-Xaa-Asp cysteine-switch motif that discriminates this protease from all other matrix metalloproteinases (MMPs) known so far. The conserved, free cysteine residue of the conventional PRCXXPD sequence interacts with the zinc ion of the catalytic domain and provides the fourth coordination site for the catalytic zinc, thereby preventing latent proMMPs from becoming active. MMPs become functionally active when proteolytic cleavage releases the prodomain and the PRCXXPD sequence and exposes the zinc atom. Here, we report that the Pro-His(81)-Cys-Gly-Xaa-Xaa-Asp motif is not functional in proMMP-26 and consequently is not involved in the activation mechanisms. Organomercurial treatment failed to activate proMMP-26. The autolytic Lys-Lys-Gln(59) downward arrow Gln(60)-Phe-His cleavage upstream of the Pro His(81)-Cys-Gly-Xaa-Xaa-Asp motif induced the proteolytic activity of recombinant proMMP-26 whereas any further cleavage inactivated the enzyme. The His(81) --> Arg(81) mutation restored the conventional cysteine-switch sequence in the prodomain but failed to induce the cysteine-switch activation mechanism. These data and computer modeling studies allowed us to hypothesize that the presence of His(81) significantly modified the fold of proMMP-26, abolished the functionality of the cysteine-switch motif, and stimulated an alternative intramolecular activation pathway of the proenzyme. PMID- 11889137 TI - Cardiomyocyte-specific gene expression following recombinant adeno-associated viral vector transduction. AB - Recombinant adeno-associated viral (rAAV) vectors hold promise for delivering genes for heart diseases, but cardiac-specific expression by the use of rAAV has not been demonstrated. To achieve this goal rAAV vectors were generated expressing marker or potentially therapeutic genes under the control of the cardiac muscle-specific alpha myosin heavy chain (MHC) gene promoter. The rAAV MHC vectors expressed in primary cardiomyocytes with similar kinetics to rAAV CMV; however, expression by the rAAV-MHC vectors was restricted to cardiomyocytes. rAAV vectors have low cytotoxicity, and it is demonstrated here that rAAV fails to induce apoptosis in cardiomyocytes compared with a recombinant adenoviral vector. rAAV-MHC or rAAV-CMV vectors were administered to mice to determine the specificity of expression in vivo. The rAAV-MHC vectors expressed specifically in cardiomyocytes, whereas the control rAAV-CMV vector expressed in heart, skeletal muscle, and brain. rAAV-MHC transduction resulted in long term (16 weeks) expression of human growth hormone following intracardiac, yet not intramuscular, injection. Finally, we defined the minimal MHC enhancer/promoter sequences required for specific and robust in vivo expression in the context of a rAAV vector. For the first time we describe a panel of rAAV vectors capable of long term cardiac specific expression of intracellular and secreted proteins. PMID- 11889138 TI - Expression of Escherichia coli glutaredoxin 2 is mainly regulated by ppGpp and sigmaS. AB - Escherichia coli glutaredoxin 2 (Grx2, encoded by grxB) differs greatly from the other two glutaredoxins in structure and catalytic properties. In a wild type strain, levels of Grx2 increased 3-fold in the stationary phase (up to 8 microg/mg). Guanosine-3',5'-tetraphoshate (ppGpp) and sigma(S), which regulate the transcription of genes in the stationary phase, dramatically affected the expression of Grx2. spoTrelA null mutants, lacking ppGpp, had very low levels of Grx2, while overproduction of full-length RelA or valine-induced starvation of isoleucine, both conditions elevating ppGpp levels, resulted in elevation of Grx2. Null mutants for the sigma(S)-specific protease ClpP, which have higher levels of sigma(S), exhibited a 3-fold Grx2 increase. sigma(S) in trans also increased the levels of Grx2. Therefore the stationary phase expression of Grx2 is determined by the sigma(S)-bound form of RNA polymerase in connection with ppGpp, while basal levels should be attributed to sigma(70)-RNA polymerase holoenzyme. Osmotic pressure and cAMP also affected the expression of Grx2, presumably via sigma(S). Furthermore, Grx2 levels were elevated in an oxyR(-) strain. In accordance with the role of Grx2 as a stationary phase protein, null mutants for grxB were shown to lyse under starvation conditions and exhibited a distorted morphology. PMID- 11889139 TI - Calcineurin-GATA-6 pathway is involved in smooth muscle-specific transcription. AB - Intracellular calcium is one of the important signals that initiates the myogenic program. The calcium-activated phosphatase calcineurin is necessary for the nuclear import of the nuclear factor of activated T cell (NFAT) family members, which interact with zinc finger GATA transcription factors. Whereas GATA-6 plays a role in the maintenance of the differentiated phenotype in vascular smooth muscle cells (VSMCs), it is unknown whether the calcineurin pathway is associated with GATA-6 and plays a role in the differentiation of VSMCs. The smooth muscle myosin heavy chain (Sm-MHC) gene is a downstream target of GATA-6, and provides a highly specific marker for differentiated VSMCs. Using immunoprecipitation Western blotting, we showed that NFATc1 interacted with GATA-6. Consistent with this, NFATc1 further potentiated GATA-6-activated Sm-MHC transcription. Induction of VSMCs to the quiescent phenotype caused nuclear translocation of NFATc1. In differentiated VSMCs, blockage of calcineurin down-regulated the amount of GATA-6 DNA binding as well as the expression of Sm-MHC and its transcriptional activity. These findings demonstrate that the calcineurin pathway is associated with GATA-6 and is required for the maintenance of the differentiated phenotype in VSMCs. PMID- 11889140 TI - Role of dynein, dynactin, and CLIP-170 interactions in LIS1 kinetochore function. AB - Mutations in the human LIS1 gene cause type I lissencephaly, a severe brain developmental disease involving gross disorganization of cortical neurons. In lower eukaryotes, LIS1 participates in cytoplasmic dynein-mediated nuclear migration. We previously reported that mammalian LIS1 functions in cell division and coimmunoprecipitates with cytoplasmic dynein and dynactin. We also localized LIS1 to the cell cortex and kinetochores of mitotic cells, known sites of dynein action. We now find that the COOH-terminal WD repeat region of LIS1 is sufficient for kinetochore targeting. Overexpression of this domain or full-length LIS1 displaces CLIP-170 from this site without affecting dynein and other kinetochore markers. The NH2-terminal self-association domain of LIS1 displaces endogenous LIS1 from the kinetochore, with no effect on CLIP-170, dynein, and dynactin. Displacement of the latter proteins by dynamitin overexpression, however, removes LIS1, suggesting that LIS1 binds to the kinetochore through the motor protein complexes and may interact with them directly. We find that of 12 distinct dynein and dynactin subunits, the dynein heavy and intermediate chains, as well as dynamitin, interact with the WD repeat region of LIS1 in coexpression/coimmunoprecipitation and two-hybrid assays. Within the heavy chain, interactions are with the first AAA repeat, a site strongly implicated in motor function, and the NH2-terminal cargo-binding region. Together, our data suggest a novel role for LIS1 in mediating CLIP-170-dynein interactions and in coordinating dynein cargo-binding and motor activities. PMID- 11889141 TI - Claudin-based tight junctions are crucial for the mammalian epidermal barrier: a lesson from claudin-1-deficient mice. AB - The tight junction (TJ) and its adhesion molecules, claudins, are responsible for the barrier function of simple epithelia, but TJs have not been thought to play an important role in the barrier function of mammalian stratified epithelia, including the epidermis. Here we generated claudin-1-deficient mice and found that the animals died within 1 d of birth with wrinkled skin. Dehydration assay and transepidermal water loss measurements revealed that in these mice the epidermal barrier was severely affected, although the layered organization of keratinocytes appeared to be normal. These unexpected findings prompted us to reexamine TJs in the epidermis of wild-type mice. Close inspection by immunofluorescence microscopy with an antioccludin monoclonal antibody, a TJ specific marker, identified continuous TJs in the stratum granulosum, where claudin-1 and -4 were concentrated. The occurrence of TJs was also confirmed by ultrathin section EM. In claudin-1-deficient mice, claudin-1 appeared to have simply been removed from these TJs, leaving occludin-positive (and also claudin-4 positive) TJs. Interestingly, in the wild-type epidermis these occludin-positive TJs efficiently prevented the diffusion of subcutaneously injected tracer (approximately 600 D) toward the skin surface, whereas in the claudin-1-deficient epidermis the tracer appeared to pass through these TJs. These findings provide the first evidence that continuous claudin-based TJs occur in the epidermis and that these TJs are crucial for the barrier function of the mammalian skin. PMID- 11889143 TI - Clinical review 142: cardiac dysrhythmias and thyroid dysfunction: the hidden menace? AB - Thyrotoxicosis is often perceived as a reversible disorder without long-term consequences, perhaps because of the availability of effective treatments, but recent evidence suggests that there may, in fact, be adverse outcomes. Long-term follow-up studies have revealed increased mortality from cardiovascular and cerebrovascular disease in those with a past history of overt hyperthyroidism treated with radioiodine as well as in those with subclinical hyperthyroidism indicated by a low serum TSH concentration. Thyroid hormones exert direct effects on the myocardium as well as the systemic vasculature predisposing to dysrhythmias, especially supraventricular. Effects of thyroid hormones on the autonomic nervous system may also contribute to arrhythmogenesis. Atrial fibrillation is a recognized complication of hyperthyroidism that predisposes to embolic events. Development of atrial fibrillation, together with other supraventricular dysrhythmias (both clinically obvious and those detected only by Holter monitoring) in those with hyperthyroidism may account for increased vascular mortality. Improved detection of supraventricular dysrhythmias and therapeutic intervention (e.g. anticoagulants, antiarrhythmics) may improve the long-term vascular prognosis, but their role remains to be established in large therapeutic trials. PMID- 11889142 TI - Osmotic stress-induced increase of phosphatidylinositol 3,5-bisphosphate requires Vac14p, an activator of the lipid kinase Fab1p. AB - Phosphatidylinositol 3,5-bisphosphate (PtdIns[3,5]P(2)) was first identified as a non-abundant phospholipid whose levels increase in response to osmotic stress. In yeast, Fab1p catalyzes formation of PtdIns(3,5)P(2) via phosphorylation of PtdIns(3)P. We have identified Vac14p, a novel vacuolar protein that regulates PtdIns(3,5)P(2) synthesis by modulating Fab1p activity in both the absence and presence of osmotic stress. We find that PtdIns(3)P levels are also elevated in response to osmotic stress, yet, only the elevation of PtdIns(3,5)P(2) levels are regulated by Vac14p. Under basal conditions the levels of PtdIns(3,5)P(2) are 18 28-fold lower than the levels of PtdIns(3)P, PtdIns(4)P, and PtdIns(4,5)P(2). After a 10 min exposure to hyperosmotic stress the levels of PtdIns(3,5)P(2) rise 20-fold, bringing it to a cellular concentration that is similar to the other phosphoinositides. This suggests that PtdIns(3,5)P(2) plays a major role in osmotic stress, perhaps via regulation of vacuolar volume. In fact, during hyperosmotic stress the vacuole morphology of wild-type cells changes dramatically, to smaller, more highly fragmented vacuoles, whereas mutants unable to synthesize PtdIns(3,5)P(2) continue to maintain a single large vacuole. These findings demonstrate that Vac14p regulates the levels of PtdIns(3,5)P(2) and provide insight into why PtdIns(3,5)P(2) levels rise in response to osmotic stress. PMID- 11889145 TI - Effects of thyroid hormone on cardiac function: the relative importance of heart rate, loading conditions, and myocardial contractility in the regulation of cardiac performance in human hyperthyroidism. PMID- 11889146 TI - In-patient hyperglycemia--are we ready to treat it yet? PMID- 11889147 TI - Hyperglycemia: an independent marker of in-hospital mortality in patients with undiagnosed diabetes. AB - Admission hyperglycemia has been associated with increased hospital mortality in critically ill patients; however, it is not known whether hyperglycemia in patients admitted to general hospital wards is associated with poor outcome. The aim of this study was to determine the prevalence of in-hospital hyperglycemia and determine the survival and functional outcome of patients with hyperglycemia with and without a history of diabetes. We reviewed the medical records of 2030 consecutive adult patients admitted to Georgia Baptist Medical Center, a community teaching hospital in downtown Atlanta, GA, from July 1, 1998, to October 20, 1998. New hyperglycemia was defined as an admission or in-hospital fasting glucose level of 126 mg/dl (7 mmol/liter) or more or a random blood glucose level of 200 mg/dl (11.1 mmol/liter) or more on 2 or more determinations. Hyperglycemia was present in 38% of patients admitted to the hospital, of whom 26% had a known history of diabetes, and 12% had no history of diabetes before the admission. Newly discovered hyperglycemia was associated with higher in hospital mortality rate (16%) compared with those patients with a prior history of diabetes (3%) and subjects with normoglycemia (1.7%; both P < 0.01). In addition, new hyperglycemic patients had a longer length of hospital stay, a higher admission rate to an intensive care unit, and were less likely to be discharged to home, frequently requiring transfer to a transitional care unit or nursing home facility. Our results indicate that in-hospital hyperglycemia is a common finding and represents an important marker of poor clinical outcome and mortality in patients with and without a history of diabetes. Patients with newly diagnosed hyperglycemia had a significantly higher mortality rate and a lower functional outcome than patients with a known history of diabetes or normoglycemia. PMID- 11889148 TI - For osteoporosis, are two antiresorptive drugs better than one? PMID- 11889149 TI - Additive effects of raloxifene and alendronate on bone density and biochemical markers of bone remodeling in postmenopausal women with osteoporosis. AB - Both raloxifene (RLX) and alendronate (ALN) can treat and prevent new vertebral fractures, increase bone mineral density (BMD), and decrease biochemical markers of bone turnover in postmenopausal women with osteoporosis. This phase 3, randomized, double-blind 1-yr study assessed the effects of combined RLX and ALN in 331 postmenopausal women with osteoporosis (femoral neck BMD T-score, less than -2). Women (aged < or = 75 yr; > or = 2 yr since their last menstrual period) received placebo, RLX 60 mg/d, ALN 10 mg/d, or RLX 60 mg/d and ALN 10 mg/d combined. At baseline, 6 and 12 months, BMD was measured by dual x-ray absorptiometry. The bone turnover markers serum osteocalcin, bone-specific alkaline phosphatase, and urinary N- and C-telopeptide corrected for creatinine were measured. The effects of RLX and ALN were considered to be independent and additive if the interaction effect was not statistically significant (P > 0.10) in a two-way ANOVA model. All changes in BMD and bone markers at 12 months were different between placebo and each of the active treatment groups, and between the RLX and RLX+ALN groups (P < 0.05). On average, lumbar spine BMD increased by 2.1, 4.3, and 5.3% from baseline with RLX, ALN, and RLX+ALN, respectively. The increase in femoral neck BMD in the RLX+ALN group (3.7%) was greater than the 2.7 and 1.7% increases in the ALN (P = 0.02) and RLX (P < 0.001) groups, respectively. The changes from baseline to 12 months in bone markers ranged from 7.1 to -16.0% with placebo, -23.8 to -46.5% with RLX, -42.3 to -74.2% with ALN, and -54.1 to -81.0% in the RLX+ALN group. RLX and ALN increased lumbar spine and femoral neck BMD, and decreased osteocalcin and C-telopeptide corrected for creatinine in an additive and independent manner, because the interaction effects were not significant. Although the ALN group had changes in BMD and bone markers that were approximately twice the magnitude as in the RLX group, it is not known how well these changes correlate to the clinical outcome of fracture. RLX+ALN reduced bone turnover more than either drug alone, resulting in greater BMD increment, but whether this difference reflects better fracture risk reduction was not assessed in this study. PMID- 11889150 TI - Infertility caused by HCG autoantibody. AB - An anti-hCG autoantibody was found in a patient with a 9-yr history of secondary infertility. Although the patient had regular menstrual cycles, had conceived spontaneously, and had good hormonal and follicular responses to gonadotropic stimulation regimens during the in vitro fertilization work-up, she presented with apparent recurrent pregnancy loss associated with prolonged raised hCG levels. Initially the presence of a high mol wt hCG complex was demonstrated in the serum by gel chromatography. The binding of [(125)I]recombinant hCG to a serum sample and subsequently to the affinity-purified IgG from the same sample revealed the presence of an hCG antibody. The antiserum was shown to be specific, with a low affinity (K(a), 1.4 x 10(6) liters/mol), but a high capacity (418 nmol/liter), for hCG. Cross-reaction with recombinant human FSH, recombinant human LH, hCG alpha, and hCG beta were low (<0.019%, 0.021%, 0.039%, and 0.006%, respectively). In addition, heat-inactivated serum and the affinity-purified IgG were shown to inhibit the action of hCG in an in vitro bioassay. We suggest that the persisting titer of the antibody to be responsible for the patient's infertility. PMID- 11889151 TI - Adrenal incidentaloma: a new cause of the metabolic syndrome? AB - A number of patients with adrenal incidentaloma are exposed to a slight degree of cortisol excess resulting from functional autonomy of the adrenal mass (usually a cortical adenoma). At present, there are only scant data on the unwanted effects of this endocrine condition referred to as subclinical Cushing's syndrome. The aim of the present study was to look for some features of the metabolic syndrome in patients with incidental adrenal adenoma. Forty-one patients (9 men and 32 women) bearing adrenal incidentaloma with typical computed tomography features of cortical adenoma were studied. For both patients and controls, exclusion criteria were age equal to 70 yr or greater, previous history of fasting hyperglycemia, or impaired glucose tolerance (IGT), severe hypertension, current use of medication or concomitant relevant illnesses, and body mass index (BMI) equal to 30 kg/m(2) or greater. Forty-one patients with euthyroid multinodular goiter accurately matched for sex, age, and BMI served for a 1:1 case-control analysis. The study design included an oral glucose tolerance test (75 g) and an endocrine workup aimed at the study of the hypothalamic-pituitary-adrenal axis. Age and BMI were fully comparable between patients (54.0 +/- 10.7 yr, 23.8 +/- 2.4 kg/m(2)) and controls (52.2 +/- 11.6 yr, 23.5 +/- 2.8 kg/m(2)). Fasting glucose and fasting insulin levels were not different between the two groups (4.96 +/- 0.61 mmol/liter vs. 4.88 +/- 0.58 mmol/liter; 67 +/- 34 pmol/liter vs. 59 +/- 32 pmol/liter), but the 2-h postchallenge glucose was significantly higher in patients than in controls (7.43 +/- 2.49 mmol/liter vs. 6.10 plus minus 1.44 mmol/liter, P = 0.01). Fifteen patients (36%) reached the World Health Organization criteria for IGT and two other patients (5%) reached those for diabetes, and 14% of the controls qualified for IGT (P = 0.01). No difference in the lipid pattern was seen between the two groups, but either systolic or diastolic blood pressure were higher in patients (135.4 +/- 15.5 mm Hg vs. 125.0 +/- 15.6 mm Hg, P = 0.003; 82.9 +/- 9.1 mm Hg vs. 75.3 +/- 6.6 mm Hg, P < 0.0001). We calculated the whole-body insulin sensitivity index derived from the oral glucose tolerance test that was significantly reduced in the patients (4.3 +/- 1.7 vs. 5.7 +/- 2.5, P = 0.01). In a multiple regression analysis, 2-h glucose was associated with BMI and midnight cortisol values (r(2) = 0.36, P = 0.002). The comparison of the patients with nonfunctioning adenoma (n = 29) with those with subclinical Cushing's syndrome (n = 12) yielded significant differences as to 2-h glucose and triglyceride levels, which were significantly higher in the second group (7.02 +/- 1.76 mmol/liter vs. 8.72 +/- 3.17 mmol/liter, P = 0.03; 1.06 +/- 0.4 mmol/liter vs. 1.73 +/- 0.96 mmol/liter, P = 0.002), but the insulin sensitivity index was conversely reduced (5.2 +/- 1.4 vs. 2.9 +/- 1.2, P < 0.0001). In conclusion, many patients with incidental adrenal adenoma display altered glucose tolerance, that may be explained by reduced insulin sensitivity, and increased blood pressure levels in comparison with carefully age- and BMI-matched controls. The slight hypercortisolism observed in some such patients may significantly contribute to this state of insulin resistance. Midnight serum cortisol appears as a sensitive marker of the metabolic effects of subclinical Cushing's syndrome. PMID- 11889152 TI - Effects of endurance and resistance training on total daily energy expenditure in young women: a controlled randomized trial. AB - There exists considerable controversy regarding the impact of different modes of exercise training on total daily energy expenditure (TEE). To examine this question, young, nonobese women were randomly assigned to a supervised 6-month program of endurance training, resistance training, or control condition. TEE was measured before and 10 d after a 6-month exercise program was completed with doubly labeled water. Body composition was determined from dual energy x-ray absorptiometry, maximum aerobic capacity from a treadmill test to exhaustion, and muscular strength from one-repetition maximum tests. Results showed that body composition did not change in endurance-trained women, but maximum aerobic capacity increased by 18%. Resistance-trained women increased muscular strength and fat-free mass (1.3 kg). TEE did not significantly change when measured subsequent to the endurance or resistance training programs. Absolute resting metabolic rate increased in resistance-trained women but not when adjusted for fat-free mass. No change in physical activity energy expenditure was found in any of the groups. These results suggest that endurance and resistance training does not chronically alter TEE in free-living young women. Thus, the energy-enhancing benefits of exercise training are primarily derived from the direct energy cost of exercise and not from a chronic elevation in daily energy expenditure in young, nonobese women. PMID- 11889153 TI - The effect of pregnancy on thyroid nodule formation. AB - Epidemiology data have revealed a higher prevalence of nodular goiters in women than men in both iodine-sufficient and iodine-deficient areas. Increased prevalence of thyroid nodules has also been reported in women with higher gravidity. However, the association between pregnancy and thyroid nodule formation has never been studied. The aim of our study was to evaluate the incidence of thyroid nodules during pregnancy and determine whether pregnancy will induce thyroid nodule formation. Two hundred twenty-one healthy southern Chinese women in the first trimester of their pregnancy were studied prospectively. Thyroid ultrasonography, thyroid function tests, and urinary iodine excretion were measured at first, second, and third trimesters of pregnancy as well as 6 wk and 3 months postpartum. Thyroid nodules (>2 mm in any dimension on ultrasonography) were detected in 34 (15.3%) subjects at first trimester, with 12 (5.4%) subjects having more than one nodule. Eight subjects had clinically palpable nodules. Women with thyroid nodules were older (P < 0.01) and had higher gravidity (P < 0.02) than those women without thyroid nodules. The volume of the single/dominant nodules increased from 60 (14--344) mm(3), median (interquartile range) at first trimester to 65 (26-472) mm(3) at third trimester (P < 0.02). These nodules remained enlarged at 103 (25-461) mm(3) 6 wk postpartum (P < 0.005) and 73 (22-344) mm(3) at 3 months postpartum (P < 0.05). Patients with thyroid nodules had lower serum TSH values (P < 0.03) and higher Tg levels (P < 0.05) throughout pregnancy. Appearance of new nodules was detected in 25 (11.3%) women as pregnancy advanced so that by 3 months postpartum, the incidence of thyroid nodular disease was 24.4% (P < 0.02 vs. first trimester). Compared with those with no detectable nodules throughout pregnancy, subjects with new nodule formation had higher urinary iodine excretion from second trimester onward (P all < 0.05). However, no difference could be detected in their TSH and Tg levels throughout pregnancy. Fine-needle aspiration on nodules greater than 5 mm in any dimension after delivery (n = 21) confirmed the majority having histological features consistent with nodular hyperplasia. No thyroid malignancy was detected. In conclusion, pregnancy is associated with an increase in the size of preexisting thyroid nodules as well as new thyroid nodule formation. This may predispose to multinodular goiter in later life. PMID- 11889154 TI - Parathyroid adenoma in a subject with familial hypocalciuric hypercalcemia: coincidence or causality? AB - A middle-aged woman presented with a history of constipation, easy fatigue, depressive mood, lassitude, polydipsia, and polyuria. The patient posed a challenging diagnostic dilemma due to the presence of persistent severe hypercalcemia and relative lack of clinically manifested symptoms. Clinical, biochemical, and genetic examinations confirmed the diagnosis of familial hypocalciuric hypercalcemia as a result of C562Y calcium-sensing receptor mutation, and a coexisting parathyroid adenoma. After adenectomy, the patient's clinical situation improved markedly, and a modest equilibrium hypercalcemia persisted. This case presents an unusual combination of two relatively common endocrine disorders. PMID- 11889155 TI - Screening for abnormal glucose tolerance in adolescents with polycystic ovary syndrome. AB - Insulin resistance is common in adults with polycystic ovary syndrome (PCOS). Although recent data demonstrate that insulin resistance is present in the early stages of PCOS, the prevalence of insulin resistance in adolescents with PCOS has not been determined. Likewise, the prevalence of impaired glucose tolerance (IGT) or type 2 diabetes mellitus (DM) in adolescent cohorts has not been established. In this study we sought to obtain preliminary data regarding the prevalence of IGT and DM in adolescents with PCOS and to assess the ability of screening tests to predict these abnormalities within this population. Twenty-seven adolescents with PCOS underwent oral glucose tolerance tests. Plasma glucose and insulin levels were obtained at baseline, and glucose was measured 2 h after a 75-g glucose challenge. The 2-h plasma glucose level was used to categorize subjects as having IGT or the provisional diagnosis of DM. Eight of our 27 subjects had IGT, and 1 had previously undiagnosed DM. These abnormalities were seen among lean and obese subjects. Fasting plasma glucose levels and simple measures of insulin resistance were suboptimal predictors of IGT and DM within our cohort. As in adults, our results indicate that adolescents with PCOS are at increased risk for IGT and DM and that the 2-h plasma glucose level after an oral glucose challenge appears to be the most reliable screening test for these abnormalities. Our results need to be corroborated by future studies that determine the prevalence of abnormalities in glucose tolerance among large populations of adolescents, both with and without PCOS. However, as DM may be preventable by lifestyle modifications, we would recommend that adolescents with PCOS undergo periodic screening for abnormal glucose tolerance using 2-h postchallenge plasma glucose levels. PMID- 11889156 TI - Parathyroid surgery: separating promise from reality. AB - We set out to determine the accuracy in predicting the success of biochemical and localizing studies for use in a minimally invasive parathyroidectomy. Preoperative sestamibi scans, intraoperative gamma-probe examinations, and intraoperative PTH (IOPTH) monitoring were performed on a prospective cohort of patients. Seventy-one patients were included in the study. Of the 59 patients (83%) with primary HPT, adenoma localization by sestamibi scanning was correct in 95% with solitary adenomas, but was correct in only 25% of the 14 patients with multiple adenomas. In patients with secondary and tertiary disease, sestamibi scanning incorrectly identified a single hot spot in 64% of cases. In no case of hyperplasia was the probe useful in locating other glands after a single gland was removed. IOPTH was accurate in 78% of patients with primary disease and in only 45% of patients with nonprimary disease. A minimal approach can be considered in a select group of patients that does not have familial primary HPT, secondary or tertiary disease, coexisting thyroid pathology, or an equivocal sestamibi scan. Only patients with a positive single hot spot on sestamibi scan can be considered candidates. Using this criteria only 64% of all patients with hyperparathyroidism are candidates for a minimally invasive approach. The combination of a solitary hot spot on sestamibi scan and a fall in IOPTH allows the surgeon to make the correct decision regarding the need to convert to a bilateral approach in 93% of these selected patients. PMID- 11889157 TI - Serum leptin level is a predictor of bone mineral density in postmenopausal women. AB - To assess whether leptin is an independent predictor of bone mineral density (BMD) in postmenopausal women, we studied the relationships of BMD to serum leptin, 25(OH)D, 1,25(OH)(2)D, PTH, E2, dehydroepiandrosterone sulfate, GH, IGF I, creatinine clearance, calcium intake, fat mass, and lean mass in 107 women aged 50-90 yr. We also related serum leptin to markers of bone formation [serum bone alkaline phosphatase and osteocalcin (OC)] and resorption (urine C telopeptide of type I collagen). In stepwise multiple linear regression, lean mass explained 28.5%, age 10.3%, and leptin 7.2% of the whole body BMD variance. Age explained 21.1%, lean mass 12.8%, and leptin 3.7% of the femoral neck BMD variance. After adjustment for fat mass and creatinine clearance, correlations between leptin and bone alkaline phosphatase (positive) and OC (negative) disappeared but, remained significant with urine C-telopeptide of type I collagen (r = -0.27, P < 0.01). Markers of bone formation and resorption were strongly intercorrelated. These data demonstrate that leptin is an independent predictor of whole body and femoral neck BMD in postmenopausal women. Although the relationships between leptin and markers of bone formation appear complex, leptin may exert a protective effect on bone by limiting the excessive bone resorption coupled to bone formation associated with bone loss after menopause. PMID- 11889158 TI - Insulin resistance, intra-abdominal fat, cardiovascular risk factors, and androgens in healthy young women with type 1 diabetes mellitus. AB - The increased cardiovascular risk in type 1 diabetes may be related, at least in part, to insulin resistance. The aim of this study was to assess the relationships between insulin sensitivity, abdominal fat, androgens, lipids, and blood pressure in 10 premenopausal women with type 1 diabetes (mean +/- SD, hemoglobin A1c 8.1 +/- 1.0%) and 10 nondiabetic body mass index-matched controls. Insulin sensitivity (glucose infusion rate during euglycemic-hyperinsulinemic clamp) was significantly less in the type 1 diabetes group than in controls (49.3 +/- 14.8 vs. 73.2 +/- 21.6 micromol/min x kg fat free mass, respectively, P = 0.01). The two groups were similar with respect to lipids, androgens, energy expenditure, physical activity, blood pressure, and abdominal adiposity (intra abdominal fat by four-slice computed tomography and central abdominal fat by dual energy x-ray absorptiometry). There were no relationships between glucose infusion rate, abdominal adiposity, and androgen levels in subjects with type 1 diabetes, in contrast to controls. Our results demonstrate greater insulin resistance in a group of premenopausal women with type 1 diabetes compared with nondiabetic controls, unrelated to abdominal adiposity, lipids, or androgens. PMID- 11889159 TI - Circulating IGF-I levels in childhood are related to both current body composition and early postnatal growth rate. AB - Rapid infancy growth predicts childhood obesity and earlier rate of maturation. We examined whether early growth rates might also influence levels of hormones relating to growth and weight gain by measuring IGF-I, IGF-II, and leptin levels in 497 normal 5-yr-old children who were followed closely from birth. IGF-I levels at 5 yr were unrelated to cord blood IGF-I levels at birth (r = 0.03; P = 0.7; n = 166) but were positively related to current weight (r = 0.32; P < 0.0005) and height (r = 0.30; P < 0.0005) and inversely related to birthweight (r = -0.21; P < 0.0005). By body composition, IGF-I levels correlated more closely with fat-free mass (r = 0.22; P < 0.0005) than with fat mass (r = 0.12; P < 0.05), whereas leptin (r = 0.57; P < 0.0005) and IGF-II levels (r = 0.15; P < 0.005) correlated more closely with fat mass. Independent of current body composition, IGF-I levels at 5 yr were significantly associated with rate of weight gain between 0-2 yr (beta = 0.19; P < 0.0005), and children who showed postnatal catch-up growth (i.e. those who showed gains in weight or length between 0-2 yr by >0.67 SD score) had higher IGF-I levels than other children (P = 0.02). IGF-II levels at 5 yr were positively related to IGF-II levels at birth (r = 0.17; P = 0.03; n = 166), and leptin levels at 5 yr were mainly related to current adiposity. Circulating IGF-I levels in childhood are influenced by infancy growth rates and possibly mediate the effects of early postnatal nutrition on later rates of growth and maturation. PMID- 11889160 TI - Complete thyroxine-binding globulin (TBG) deficiency in two families without mutations in coding or promoter regions of the TBG genes: in vitro demonstration of exon skipping. AB - Inherited thyroxine-binding globulin (TBG) deficiency is caused by mutations in the TBG gene located on the X-chromosome. We now describe two families (K and H) with X-linked complete TBG deficiency without mutations in the coding or promoter regions of the TBG gene. The propositi of both families presented with euthyroid hypothyroxinemia and were found to have undetectable TBG in serum. Affected females had approximately half the normal serum TBG concentration except for one woman who also had undetectable TBG (family H). All four of her children (two boys and two girls) were affected. Affected members of family K had no mutations in any of the five exons or in the minimal promoter region of the TBG gene. However, a G to A substitution, five base pairs downstream from exon 3, was associated to the phenotype of TBG deficiency (TBG-Jackson) and was not present in 100 normal alleles. In contrast to individuals without this mutation, no TBG mRNA could be detected in fibroblasts of the propositus, expressing solely TBG Jackson. In vitro transcription of genomic DNA containing the mutant intron in an exon trapping system showed that this mutation, reducing the consensus value on the 5' donor splice site, affects the normal splicing process. The transcript of TBG-Jackson lacks exon 3 and is unstable. The deduced amino acid sequence has a frameshift and an early stop codon at position 325. Affected subject of family H had no mutations in the TBG gene including all exons, all introns, the minimal promoter, and the 3' untranslated sequence. However, an intragenic A/G polymorphism (125 bp upstream from exon 2) was identified. It allowed us to confirm a cosegregation of the phenotype to the TBG gene and to show that the single female with complete TBG deficiency was homozygous for the polymorphic TBG allele. The cause of TBG deficiency in this family remains unknown. PMID- 11889161 TI - Male LH-independent sexual precocity in a 3.5-year-old boy caused by a somatic activating mutation of the LH receptor in a Leydig cell tumor. AB - We describe the clinical features of severe sexual precocity in a 3.5-yr-old boy. Hormonal evaluation showed LH-independent T hypersecretion. Initial examination of the adrenals and testes revealed no evidence of congenital adrenal hyperplasia, hCG- or androgen-secreting tumors, or McCune-Albright syndrome. In the coding sequence of the LH receptor gene no activating mutation was found. Spironolactone (5.7 mg/kg x d) and testolactone (40 mg/kg x d) were unsuccessful in suppressing the elevated concentration of T. To further determine the origin of the elevated serum T, a selective venous sampling procedure was planned. However before the sampling procedure, high resolution ultrasound examination showed a small tumor in the left testis, which was removed. Histology proved the tumor to be a Leydig cell adenoma. Sequencing of the tumor LH receptor gene revealed a heterozygous mutation in exon 11 encoding a replacement of aspartic acid at position 578 with histidine, which has been shown to be a constitutively activating mutation. These findings indicate that in male patients with gonadotropin-independent sexual precocity, the presence of small testicular Leydig cell tumors harboring a somatic mutation of the LH receptor gene should be considered. PMID- 11889162 TI - Two de novo mutations in the AR gene cause the complete androgen insensitivity syndrome in a pair of monozygotic twins. AB - The androgen insensitivity syndrome (AIS) is the most common cause of male undermasculinization and is typically caused by mutations in the AR gene. Affected individuals may exhibit either complete external feminization (complete AIS) or a partial phenotype (partial AIS). Here we describe monozygotic twins diagnosed with complete AIS who each possess two substitutions (C-->G at position 2930 and T-->C at position 2955, both in exon 7), leading to Phe(856)Leu and Ser(865)Pro mutations, respectively. Neither parent was found to be a carrier for these mutations, indicating that the double mutation arose de novo. Both mutations were recreated by site-directed mutagenesis and compared functionally with the wild-type receptor. The Phe(856)Leu mutation did not affect androgen binding when expressed in COS-1 cells, nor did this mutation decrease androgen dependent trans-activation in transfected HeLa cells. However, the Ser(865)Pro mutation completely ablated androgen binding and trans-activation. In this study we demonstrate that the replacement of serine by proline at position 865 is sufficient in itself to cause complete AIS in these twins. Analyses of nuclear receptor structures suggest that this mutation is likely to perturb the conformation of helix 10/11, which plays a role in ligand binding, dimerization, and receptor activation. To our knowledge this is the first confirmed instance of AIS (complete or partial) due to an AR mutation occurring in twins. Furthermore, the phenotype was associated with two mutations that were both novel in nature. PMID- 11889163 TI - Estrogen status correlates with the calcium content of coronary atherosclerotic plaques in women. AB - BACKGROUND: Coronary artery calcium, a radiographic marker for atherosclerosis and a predictor of coronary heart disease (CHD), is less extensive in women than in men of the same age. The role of estrogen in the pathogenesis of coronary artery calcification is unknown. We examined the association of estrogen status with extent of calcification and atherosclerotic plaque in coronary arteries of deceased women. METHODS: Coronary arteries were obtained at autopsy from 56 white women age 18--98 yr, 46 postmenopausal and 10 premenopausal. Exclusion criteria included patients with coronary stents, coronary artery bypass surgery, and medical-legal cases. Medical records were reviewed for demographics, CHD risk factors, menstrual status, and use of estrogen replacement therapy. Contact microradiography of coronary arteries assessed true calcium content and atherosclerotic plaque area was analyzed histologically. RESULTS: The coronary arteries from estrogen-treated postmenopausal women had lower mean coronary calcium content (P = 0.002), mean plaque area (P < 0.0001), and calcium-to-plaque area ratio (P = 0.004) than those from untreated menopausal women. Estrogen status, age, diabetes, and hypertension predicted calcium and plaque area by univariate analysis. After controlling for these CHD risk factors, estrogen status remained an independent predictor of both calcium (P = 0.014) and plaque area (P = 0.001) in all women. Mean calcium area (P < 0.05) but not plaque area (P = 0.44) was significantly greater in women treated with estrogen replacement therapy than in premenopausal women. Coronary calcium (P < 0.007) and plaque area (P < 0.03) varied significantly with age in untreated postmenopausal women, but not in the estrogen-treated or premenopausal women (P = 0.33). CONCLUSIONS: Estrogen status is associated with coronary calcium and plaque area independent of age and CHD risk factors. Estrogen may modulate the calcium content of atherosclerotic plaques, as well as plaque area and may slow the progression of atherosclerosis in women. PMID- 11889165 TI - Narrow individual variations in serum T(4) and T(3) in normal subjects: a clue to the understanding of subclinical thyroid disease. AB - High individuality causes laboratory reference ranges to be insensitive to changes in test results that are significant for the individual. We undertook a longitudinal study of variation in thyroid function tests in 16 healthy men with monthly sampling for 12 months using standard procedures. We measured serum T(4), T(3), free T(4) index, and TSH. All individuals had different variations of thyroid function tests (P < 0.001 for all variables) around individual mean values (set points) (P < 0.001 for all variables). The width of the individual 95% confidence intervals were approximately half that of the group for all variables. Accordingly, the index of individuality was low: T(4) = 0.58; T(3) = 0.54; free T(4) index = 0.59; TSH = 0.49. One test result described the individual set point with a precision of +/- 25% for T(4), T(3), free T(4) index, and +/- 50% for TSH. The differences required to be 95% confident of significant changes in repeated testing were (average, range): T(4) = 28, 11-62 nmol/liter; T(3) = 0.55, 0.3--0.9 nmol/liter; free T4 index = 33, 15-61 nmol/liter; TSH = 0.75, 0.2-1.6 mU/liter. Our data indicate that each individual had a unique thyroid function. The individual reference ranges for test results were narrow, compared with group reference ranges used to develop laboratory reference ranges. Accordingly, a test result within laboratory reference limits is not necessarily normal for an individual. Because serum TSH responds with logarithmically amplified variation to minor changes in serum T(4) and T(3), abnormal serum TSH may indicate that serum T(4) and T(3) are not normal for an individual. A condition with abnormal serum TSH but with serum T(4) and T(3) within laboratory reference ranges is labeled subclinical thyroid disease. Our data indicate that the distinction between subclinical and overt thyroid disease (abnormal serum TSH and abnormal T(4) and/or T(3)) is somewhat arbitrary. For the same degree of thyroid function abnormality, the diagnosis depends to a considerable extent on the position of the patient's normal set point for T(4) and T(3) within the laboratory reference range. PMID- 11889166 TI - High dose of (131)I therapy for the treatment of hyperthyroidism caused by Graves' disease. AB - Radioactive iodine ((131)I) has become the most widely used therapy for patients with hyperthyroidism caused by Graves' disease in the United States. There remains, however, significant variability among (131)I dosing regimens, and it is clear that most patients ultimately develop hypothyroidism after therapy. To avoid persistent hyperthyroidism, we adopted a high dose (131)I therapy protocol based on measurement of 24-h thyroid (123)I uptake designed to deliver 8 mCi (296 MBq) to the thyroid gland 24 h after (131)I administration. To evaluate the efficacy of this protocol, we reviewed our clinical experience over a 7-yr period. We treated 261 patients (219 women and 42 men) with hyperthyroidism caused by Graves' disease with (131)I [mean dose, 14.6 mCi (540 MBq)] between 1993 and 1999. Before treatment, 207 (79%) had received an antithyroid drug (109 propylthiouracil and 98 methimazole). We determined their thyroid status 1 yr after treatment in relation to age, pretreatment with an antithyroid drug, pretreatment thyroid size, and dose of (131)I retained in the thyroid 24 h after treatment. Among the 261 patients, 225 (86%) were euthyroid or hypothyroid 1 yr after treatment, and 36 patients (14%) had persistent hyperthyroidism and required a second treatment. The patients who had persistent hyperthyroidism were younger (P < 0.01), had larger thyroid glands (P < 0.01), higher pretreatment thyroid (123)I uptake values (P < 0.01), and higher serum T(4) concentrations (P < 0.01) and were more likely to have taken antithyroid medication before administration of (131)I (P = 0.01). Five of these patients developed transient hypothyroidism, followed by thyrotoxicosis. There was an asymptotic, inverse relationship between the retained dose of (131)I at 24 h and persistent hyperthyroidism, revealing a 5-10% failure rate despite delivery of up to 400 microCi (14.8 MBq)/g. A dose of (131)I that results in accumulation of 8 mCi (296 MBq) in the thyroid gland 24 h after administration is an effective treatment for the majority of patients with Graves' hyperthyroidism. Young patients with larger thyroid glands, higher serum T(4) concentrations, and higher 24-h thyroid (123)I uptake values, and those pretreated with antithyroid medication for greater than 4 months are at higher risk for treatment failure. A higher dose of (131)I may be advisable in such patients. PMID- 11889168 TI - Loss of body cell mass in Cushing's syndrome: effect of treatment. AB - Cushing's syndrome (CS) is associated with low fat-free mass, but it is unclear whether hypercortisolism causes a loss of whole body protein. Body composition was studied prospectively in 15 patients with untreated CS (n = 14 pituitary adenoma; n = 1 adrenal adenoma), in 15 nonobese healthy controls, and in 15 weight-matched obese controls by 3 different methods: total body potassium counting (TBP), bioelectrical impedance analysis (BIA), and anthropometry. In 6 patients, body composition was studied before and within 6 months after pituitary surgery. In CS patients and weight-matched controls, body weight and total body fat were significantly higher than in nonobese controls. In CS patients, TBP was 18.4% lower than predicted, whereas in weight-matched controls TBP was 7.1% higher than predicted. As compared with nonobese and weight-matched controls, in CS patients TBP indicated a significant loss of body cell mass (BCM) of -20.2 and -21.1%, respectively. A significantly reduced arm muscle area of -21.3% compared with weight-matched controls also indicated a loss of whole body protein. In CS, however, BIA overestimated BCM when compared with TBP by +18% and agreement between BIA and TBP in the individual patient was poor (limits of agreement plus minus 27.6%), indicating the invalidity of standard BIA equations in this population. Measurements performed before and 6 months after successful pituitary surgery demonstrated a significant loss of body weight (-11%) and body fat ( 33%), but BCM and muscle mass remained on a constant low level. In conclusion, this study shows that, in patients with CS, a significantly reduced BCM indicates a true protein loss. The second interesting finding is that in the early recovery after successful treatment of hypercortisolism patients lose body fat without gaining BCM or muscle mass. PMID- 11889169 TI - Serum leptin concentrations in endometriosis. AB - It has been recently reported that serum concentrations of the adipocyte-derived hormone leptin are increased in patients affected by endometriosis. On the basis of these findings, the present study was undertaken to evaluate whether the protein may be used as a new serum marker of the disease. A consecutive series of 67 reproductive-age women who underwent laparoscopy for benign gynecological pathologies were enrolled prospectively for the study. Serum leptin concentrations, as evaluated by a conventional RIA kit, were related to baseline clinical characteristics and surgical and histologic diagnosis. Endometriosis was documented in 42 women (stage I-II in 19 patients and stage III-IV in 23 patients). Twenty-five women of similar age and body mass index, who had no laparoscopic evidence of the disease, served as control group. Serum levels of leptin resulted similar between women without and with endometriosis at any stage (mean +/- SEM, 12.5 +/- 9.4 ng/ml and 12.1 +/- 8.0 ng/ml, respectively). No significant association with leptin concentrations was observed in regard to stage of the disease, number of endometriotic implants, presence/absence of an endometriotic cyst or peritoneal deep endometriosis, and presence/absence of specific symptoms. Therefore, our results do not support the possibility to employ leptin measurement as a diagnostic tool for endometriosis. Further studies are needed to elucidate the relationship between leptin and endometrial system and determine the potential contribution of the molecule in implantation and early pregnancy development. PMID- 11889170 TI - The cardiovascular risk of adult GH deficiency (GHD) improved after GH replacement and worsened in untreated GHD: a 12-month prospective study. AB - Increased cardiovascular morbidity and mortality were reported in GH deficiency (GHD), and GH replacement can ameliorate cardiac abnormalities of adult GHD patients. To test the potential progression of untreated GHD on the cardiovascular risk and cardiac function, cardiovascular risk factors, cardiac size, and performance were prospectively evaluated in 15 GHD patients (age, 18-56 yr) who were treated with recombinant GH at the dose of 0.15-1.0 mg/d, 15 GHD patients (age, 18-56 yr) who refused GH replacement, and 30 healthy subjects (age, 18-53 yr). Electrocardiogram, systolic and diastolic blood pressure, and heart rate measurement, serum IGF-I, total cholesterol, low- and high-density lipoprotein (LDL, HDL) cholesterol, triglycerides, and fibrinogen level assay, echocardiography, and equilibrium radionuclide angiography were performed basally and after 12 months. At study entry, low IGF-I levels, unfavorable lipid profile, and inadequate cardiac and physical performance were found in GHD patients compared with controls. After 12 months of GH treatment, IGF-I levels normalized; HDL-cholesterol levels, left ventricular (LV) mass index (LVMi), left ventricular ejection fraction (LVEF) at peak exercise, peak filling rate, exercise duration and capacity significantly increased; total- and LDL-cholesterol levels significantly decreased. After 12 months in GH-untreated GHD patients, IGF-I levels remained stable, and HDL-cholesterol levels, LVEF both at rest and at peak exercise, and exercise capacity were further reduced; total- and LDL-cholesterol levels increased slightly. LVEF at rest and its response at peak exercise normalized in 60 and 53.3%, respectively, of GH-treated patients and in none of the GH-untreated patients. In conclusion, 12 months of GH replacement normalized IGF-I and improved lipid profile and cardiac performance in adult GHD patients. A similar period of GH deprivation induced a further impairment of lipid profile and cardiac performance. This finding strongly supports the need of GH replacement in adult GHD patients. PMID- 11889171 TI - Identification of suppressors of cytokine signaling (SOCS) proteins in human gestational tissues: differential regulation is associated with the onset of labor. AB - Inflammatory cytokines secreted by the placenta and fetal membranes are believed to play an important role in the initiation of parturition. The suppressor of cytokine signaling (SOCS) proteins regulate signal transduction by several cytokines that have been reported to affect gestational tissues. The presence, distribution and roles of SOCS proteins, however, have not been described in human gestational tissues. Using reverse transcriptase (RT)-PCR and Western blot analysis we investigated the expression of SOCS1, SOCS2, and SOCS3 mRNA and protein, respectively, by human villous placenta, amnion and choriodecidua (n = 3 4). Tissues were obtained from uncomplicated pregnancies at term after either spontaneous labor and vaginal delivery or caesarean section (before labor). Messenger RNAs for SOCS1, SOCS2, and SOCS3 were expressed in all tissue types, irrespective of labor status. SOCS proteins were, however, only detectable in villous placenta and in one case in the choriodecidua. Labor was associated with abrogated expression of SOCS1 and SOCS3 proteins in villous placenta and the choriodecidua sample. Following labor the band for SOCS2 protein increased slightly in size which may indicate post-translational modification of SOCS2. Reduced expression of SOCS proteins in gestational tissues may provide a mechanism by which inflammatory cytokines enter into a positive feedback loop of inflammatory changes leading to delivery. PMID- 11889172 TI - Stimulation of autologous antitumor T-cell responses against medullary thyroid carcinoma using tumor lysate-pulsed dendritic cells. AB - Dendritic cells (DCs) have attracted wide interest because of their unique capacity to elicit primary and secondary antitumor responses. We have generated autologous tumor lysate-pulsed DCs from three patients with medullary thyroid carcinoma (MTC) and tested them for their ability to stimulate cytotoxic T-cell responses against autologous MTC tumor cells in vitro. The aim of our investigations was to evaluate the potential efficacy of DC-based immunotherapy in patients with MTC. DCs were generated from peripheral blood monocytes using GM CSF and IL-4 (immature DCs) or GM-CSF, IL-4, and TNFalpha (mature DCs). Our results indicate that mature tumor lysate-pulsed DCs are able to elicit a human leukocyte antigen class I-restricted cytotoxic T-cell response against autologous MTC tumor cells, whereas immature tumor lysate-pulsed DCs do not stimulate significant antitumor activity. We feel that our data may be relevant for future clinical trials of active immunotherapy using tumor lysate-pulsed DCs in patients with MTC who have residual or distant disease after surgical treatment. The fact that mature DCs displayed a substantially higher capacity to stimulate autologous antitumor T-cell responses than immature DCs underlines the importance of a maturation step in immunotherapy protocols based on DCs. PMID- 11889173 TI - The activity of PPAR gamma in primary human trophoblasts is enhanced by oxidized lipids. AB - The ligand-dependent nuclear receptor PPAR gamma plays an important role in murine and human trophoblast differentiation. Oxidized lipids, which are implicated in the pathophysiology of placental dysfunction, have recently been identified as ligands for PPAR gamma. We therefore hypothesized that oxidized lipids activate PPAR gamma in human trophoblasts and influence placental function. To test our hypothesis, we examined the effect of 9S-hydroxy-10E,12Z octadecadienoic acid (9-HODE), 13S-hydroxy-9Z,11E-octadecadienoic acid (13-HODE), and 15S-hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic acid (15-HETE) on PPAR gamma activity in cultured term human trophoblasts. Our results demonstrate that these lipids stimulate PPAR gamma activity and that the AF-2 fragment, which harbors the ligand-binding domain of PPAR gamma, mediates this effect. Furthermore, we assessed the consequences of PPAR gamma activation by the oxidized lipids, and we found that these lipids stimulate human CG production, a measure of trophoblast differentiation. In contrast, the expression of syncytin, a marker for syncytium formation as well as the expression of the cell cycle modulators cyclin E and p27 are unchanged by the oxidized lipids. We concluded that 9-HODE, 13-HODE, and 15 HETE activate PPAR gamma in primary human trophoblasts. These PPAR gamma ligands may play a role in placental differentiation, yet they are unlikely to contribute to trophoblast dysfunction. PMID- 11889174 TI - Impaired developmental competence of oocytes in adult prenatally androgenized female rhesus monkeys undergoing gonadotropin stimulation for in vitro fertilization. AB - To determine whether prenatal T propionate exposure beginning gestational d 40-44 (early-treated) or 100-115 (late-treated) affects oocyte competence, five early treated and five late-treated prenatally androgenized and five normal monkeys underwent recombinant human FSH injections with oocyte-retrieval after hCG administration. Serum FSH, LH, estradiol (E(2)), progesterone (P(4)), androstenedione (A(4)), T, and dihydrotestosterone were measured basally, during gonadotropin stimulation and at oocyte-retrieval; fasting serum glucose and insulin also were determined basally and at oocyte-retrieval. Follicle fluid sex steroids were analyzed. Oocyte number, nuclear maturity, and fertilization were comparable among female groups, but the percentage of zygotes developing into blastocysts was reduced in early-treated prenatally androgenized females. The intrafollicular P(4)/E(2) ratio was significantly elevated in early-treated prenatally androgenized females, whereas intrafollicular P(4)/A(4) and T/A(4) ratios were significantly increased in all prenatally androgenized females. Early treated prenatally androgenized females demonstrated persistent LH hypersecretion. They also were unable to suppress circulating insulin levels during gonadotropin stimulation. Circulating sex steroid levels and serum P(4)/E(2), P(4)/A(4), and E(2)/androgen ratios were similar in all females. Early prenatal androgenization in monkeys receiving gonadotropins impairs oocyte developmental competence and seems to induce premature follicle differentiation in the presence of LH hypersecretion and relative insulin excess. PMID- 11889175 TI - Functionally impaired TR mutants are present in thyroid papillary cancer. AB - TRs are transcription factors that regulate cell proliferation, differentiation, and apoptosis. They are cellular homologs of the transcriptionally inactive viral oncogene v-erbA. We tested the hypothesis that the functions of TRs could be impaired in cancer tissues as a result of aberrant expression and/or somatic mutations. As a model system, we selected human thyroid papillary cancer, in which the most common abnormalities, RET/papillary thyroid cancer rearrangements (fusion of RET kinase domain to the activating domains of other genes), were found in 40--45% of cases. We found that the mean expression levels of TR beta mRNA and TR alpha mRNA were significantly lower, whereas the protein levels of TR beta 1 and TR alpha 1 were higher in cancer tissues than in healthy thyroid. Sequencing of TR beta 1 and TR alpha 1 cDNAs, cloned from 16 papillary cancers, revealed that mutations affected receptor amino acid sequences in 93.75% and 62.5% of cases, respectively. In contrast, no mutations were found in healthy thyroid controls, and only 11.11% and 22.22% of thyroid adenomas had such TR beta 1 or TR alpha 1 mutations, respectively. The majority of the mutated TRs lost their trans-activation function and exhibited dominant negative activity. These findings suggest a possible role for mutated thyroid hormone receptors in the tumorigenesis of human papillary thyroid carcinoma. PMID- 11889177 TI - The -597 G-->A and -174 G-->C polymorphisms in the promoter of the IL-6 gene are associated with hyperandrogenism. AB - To evaluate whether genetic variability at the IL-6 gene (IL-6) is associated with hyperandrogenism, we studied four common polymorphisms in the IL-6 promoter (-597G-->A, -572G--> C, -373A(n)T(n), -174G-->C) in 85 hyperandrogenic patients and 25 healthy women. We found 5 different haplotypes when considering the 3 biallelic polymorphisms at positions -597, -572, and -174 of IL-6 (relative frequencies in parentheses): GGG (0.505), AGC (0.377), GGC (0.059), GCG (0.055), and GCC (0.005). The frequencies of the GGG haplotype were 0.559 in patients and 0.320 in controls, whereas those of the AGC haplotype were 0.318 in patients and 0.580 in controls (chi(2) = 12.145; P < 0.02). The -597G-->A and -174G-->C polymorphisms were in linkage disequilibrium (chi(2) = 152.220; P < 0.00001), and were associated with patient or control status. -597G and -174G alleles were more frequent in patients in homozygosity or considering subjects homozygous and heterozygous for G alleles as a whole (P < 0.05 for all analyses). In healthy women G alleles at -597 and -174 were associated with statistically significant higher circulating levels of IL-6 and basal cortisol, 11-deoxycortisol, and 17 hydroxyprogesterone and a tendency (P < 0.10) for higher total T concentrations compared with -597A and -174C alleles. On the contrary, neither the -572G-->C nor the -373A(n)T(n) polymorphism was related to hyperandrogenism or influenced any clinical or biochemical variable. In conclusion, our present results suggest that the -597G-->A and -174G-->C polymorphisms in IL-6 are involved in the pathogenesis of hyperandrogenic disorders. PMID- 11889176 TI - Troglitazone, an insulin-sensitizing thiazolidinedione, represses combined stimulation by LH and insulin of de novo androgen biosynthesis by thecal cells in vitro. AB - Polycystic ovarian syndrome (anovulatory hyperandrogenism) is marked by adolescent onset of systemic hyperinsulinism, oligoovulation, hirsutism, excessive LH and androgen secretion, and variable reduction in fertility. Insulin and LH are believed to act in concert to promote ovarian androgen hypersecretion in this disorder. Administration of troglitazone, an insulin-sensitizing agent and putative PPAR gamma agonist, can decrease hyperinsulinism, suppress T production, and ameliorate oligoovulation in some women with this endocrinopathy. The present study tests the hypothesis that troglitazone directly inhibits de novo androgen biosynthesis stimulated jointly by LH and insulin in primary cultures of (porcine) thecal cells. We show that troglitazone dose-dependently antagonizes LH/insulin's combined stimulation of androstenedione and T production by thecal cells in vitro. Consistent steroidogenic inhibition of 80-95% was achieved at drug concentrations of 3-6.8 microM (P < 0.001). Exposure of thecal cells to the thiazolidinedione derivative also blocked bihormonally stimulated accumulation of CYP17 (cytochrome P450 17 alpha-hydroxylase/C(17-20) lyase) gene expression, as reflected by decreased accumulation of cognate heterogeneous nuclear RNA and mRNA (by 30-65%; P < 0.05). Moreover, troglitazone suppressed LH/insulin-induced phosphorylation of the 52-kDa immunoprecipitated CYP17 enzyme by 88% (P < 0.001). A putative natural agonist of PPAR gamma nuclear transcription, 15-deoxy-delta-12,14-prostaglandin J(2), also inhibited LH/insulin driven androstenedione biosynthesis and CYP17 gene expression in thecal cells. In conclusion, a synthetic thiazolidinedione (troglitazone) and a natural ligand of PPAR gamma (15-deoxy-delta-12,14-prostaglandin J(2)) effectively impede the concerted stimulation by LH and insulin of in vitro thecal cell androgen production, CYP17 gene expression, and CYP17 protein phosphorylation. This ensemble of inhibitory actions on LH/insulin-stimulated steroidogenesis offers a plausible mechanistic basis for at least part of the observed clinical efficacy of troglitazone in mitigating androgen excess in women with polycystic ovarian syndrome. PMID- 11889178 TI - Potent inhibition of estrogen sulfotransferase by hydroxylated metabolites of polyhalogenated aromatic hydrocarbons reveals alternative mechanism for estrogenic activity of endocrine disrupters. AB - Polyhalogenated aromatic hydrocarbons (PHAHs), such as polychlorinated dibenzo-p dioxins and dibenzofurans, polybrominated diphenylethers, and bisphenol A derivatives are persistent environmental pollutants, which are capable of interfering with reproductive and endocrine function in birds, fish, reptiles, and mammals. PHAHs exert estrogenic effects that may be mediated in part by their hydroxylated metabolites (PHAH-OHs), the mechanisms of which remain to be identified. PHAH-OHs show low affinity for the ER. Alternatively, they may exert their estrogenic effects by inhibiting E2 metabolism. As sulfation of E2 by estrogen sulfotransferase (SULT1E1) is an important pathway for E2 inactivation, inhibition of SULT1E1 may lead to an increased bioavailability of estrogens in tissues expressing this enzyme. Therefore, we studied the possible inhibition of human SULT1E1 by hydroxylated PHAH metabolites and the sulfation of the different compounds by SULT1E1. We found marked inhibition of SULT1E1 by various PHAH-OHs, in particular by compounds with two adjacent halogen substituents around the hydroxyl group that were effective at (sub)nanomolar concentrations. Depending on the structure, the inhibition is primarily competitive or noncompetitive. Most PHAH-OHs are also sulfated by SULT1E1. We also investigated the inhibitory effects of the various PHAH-OHs on E2 sulfation by human liver cytosol and found that the effects were strongly correlated with their inhibitions of recombinant SULT1E1 (r = 0.922). Based on these results, we hypothesize that hydroxylated PHAHs exert their estrogenic effects at least in part by inhibiting SULT1E1 catalyzed E2 sulfation. PMID- 11889179 TI - A Novel mutation in the FSH receptor inhibiting signal transduction and causing primary ovarian failure. AB - Inactivating mutations of the FSH receptor (FSHR) are known to cause ovarian failure with amenorrhea and infertility in women. The first mutation identified in the FSHR gene was a missense mutation (566C-->T, predicting Ala189Val transition) found in several Finnish patients with primary amenorrhea due to ovarian failure. Only five additional, partially or totally inactivating, mutations of the FSHR have been reported. Here, we report a novel FSHR mutation, 1255G-->A, in a Finnish female with primary amenorrhea. The patient was a compound heterozygote for two mutations in the FSHR gene: 566C-->T, the Finnish founder mutation, and 1255G-->A, a previously unidentified mutation. The new mutation is located in exon 10 in the second transmembrane stretch of the FSHR, and it predicts an Ala419Thr change in the protein structure. In functional testing, the mutation was shown to have minimal effect on ligand binding capacity and affinity, but it almost totally abolished the cAMP second messenger response. Neither of the two FSHR mutations (566C-->T or1255G-->A) was identified in 40 other Finnish patients with premature ovarian failure. Based on this and previous studies, FSHR mutations remain a rare cause of ovarian failure. PMID- 11889180 TI - Stimulation and growth of antral ovarian follicles by selective LH activity administration in women. AB - Intensive FSH stimulation is a key tool of assisted reproduction technology but can cause severe complications through the development of an excessive number of small ovarian follicles. We tested the hypothesis that, in the late stages of ovulation induction, LH activity in the form of low-dose human CG (hCG) can stimulate and selectively modulate ovarian follicle function and growth, independently of FSH administration. Four groups of GnRH agonist-suppressed normoovulatory women (10 each group) received recombinant human FSH (r-hFSH) (150 IU/d) for 7 d followed by: group A, r-hFSH 150 IU/d alone; group B, r-hFSH 50 IU/d and hCG 50 IU/d; group C, r-hFSH 25 IU/d and hCG 100 IU/d; group D, hCG 200 IU/d alone. Despite several days of lowered or absent r-hFSH administration, 70% of hCG-treated patients successfully completed treatment. In these subjects, preovulatory E2 levels and large (>14 mm diameter) ovarian follicle development were not reduced; conversely, the number of small (<10 mm diameter) ovarian follicles was significantly decreased in groups B-D vs. group A. Low-dose hCG administration did not cause follicle luteinization. We conclude that, following FSH priming, LH activity administration can: 1) stimulate folliculogenesis for several days, in spite of rapidly declining FSH levels; and 2) hasten small follicle demise. Therefore, LH activity administration could be used to design radically novel ovulation induction regimens that, by partly or completely replacing mid-/late follicular phase FSH administration, may reduce costs and improve safety of assisted reproduction technology. PMID- 11889181 TI - Relationship between the GH/IGF-I axis, insulin sensitivity, and adrenal androgens in normal prepubertal and pubertal boys. AB - In girls, but not in boys, pronounced adrenarche and precocious pubarche along with ovarian hyperandrogenism have been related to insulin resistance and reduced fetal growth. However, insulin secretion is increased during puberty in normal boys. The aim of this study was to analyze the possible implication of changes in the GH/IGF-I axis and in insulin sensitivity for the regulation of adrenal androgen secretion of normal prepubertal and adolescent boys. Fifty-six normal boys were divided into the following groups (Gr): Gr1, prepuberty (testicular volume, <4 cc; n = 33); and Gr3, puberty (testicular volume, 4-25 cc; n = 23). Gr1 was subdivided according to age into: Gr1A, early prepuberty (boys younger than 5.9 yr old; n = 16); and Gr1B, late prepuberty (prepubertal boys, 5.9 yr old or older; n = 17). Gr3 was subdivided according to testicular volume into: Gr3A, early puberty (testicular volume, 4-8 cc; n = 13); and Gr3B, late puberty (testicular volume, 10--25 cc; n = 10). To study hormonal changes during the transition between prepuberty and puberty, an additional group, Gr2 (n = 30), was defined by mixing Gr1B and Gr3A. Serum dehydroepiandrosterone sulfate (DHEAS), androstenedione (Delta(4)A), insulin, IGF-I, and glucose were determined after overnight fasting. Insulin sensitivity was estimated by the fasting glucose/insulin (G/I) ratio. There was a close correlation between fasting G/I ratio and QUICKI, a quantitative insulin sensitivity check index. Mean values for Gr1 and Gr3 as well as their subgroups were compared using t test. In Gr1, the mean fasting G/I ratio was significantly higher, and the mean serum IGF-I, serum DHEAS, and serum Delta(4)A levels were significantly lower than in Gr3 (P < 0.001). Mean fasting G/I ratios in Gr1A and Gr3A were not significantly different from those in Gr1B and Gr3B, respectively, but the fasting G/I ratio in Gr3A was significantly lower than that in Gr1B (P < 0006). Moreover, body mass index (BMI) in Gr3A was significantly higher than that in Gr1B (P < 0.01). On the other hand, mean serum IGF-I levels in Gr1A and Gr3A were significantly lower than those in Gr1B and Gr3B, respectively (P < 0.0001). The mean serum DHEAS level in Gr1A was significantly lower than that in Gr1B (P < 0.01), but no difference was found between Gr3A and Gr3B. The mean serum Delta(4)A in Gr1A was similar to that in Gr1B, but the mean serum Delta(4)A in Gr3A was significantly lower than that in Gr3B (P = 0.0001). Correlation studies within Gr1, Gr2, and Gr3 were also carried out. There was a significant positive correlation between serum DHEAS and age in Gr1 and Gr2, but not in Gr3. In Gr1, no significant correlation was found between serum DHEAS and fasting G/I ratio or between serum DHEAS and serum IGF-I, suggesting that adrenal steroidogenesis in male prepuberty is independent of insulin sensitivity or peripheral IGF-I. In Gr2, a significant negative correlation (P = 0.01) between serum DHEAS and the fasting G/I ratio was found, but not between serum DHEAS and serum IGF-I. Furthermore, a significant negative correlation between BMI and the fasting G/I ratio was also found. Therefore, changes in insulin sensitivity might be involved in adrenal androgen synthesis during the transition from prepuberty to puberty. Finally, in Gr3, DHEAS was not significantly correlated with the fasting G/I ratio or serum IGF-I. A significant negative correlation between serum Delta(4)A and the fasting G/I ratio was found in Gr2. In Gr2, but not in Gr3, there was a significant negative correlation between the fasting G/I ratio and age (P = 0.03) and between the fasting G/I ratio and serum IGF-I (P = 0.03). In conclusion, our data support the hypothesis that the GH/IGF-I axis and insulin sensitivity are not involved in the mechanism of adrenarche in boys. Insulin sensitivity and BMI, however, decrease at early puberty rather than at late puberty, and this change could be involved in modulating adrenal androgen steroidogenesis during the transition between late prepuberty and early puberty. PMID- 11889182 TI - Association of H19 promoter methylation with the expression of H19 and IGF-II genes in adrenocortical tumors. AB - Low H19 and abundant IGF-II expression may have a role in the development of adrenocortical carcinomas. In the mouse, the H19 promoter area has been found to be methylated when transcription of the H19 gene is silent and unmethylated when it is active. We used PCR-based methylation analysis and bisulfite genomic sequencing to study the cytosine methylation status of the H19 promoter region in 16 normal adrenals and 30 pathological adrenocortical samples. PCR-based analysis showed higher methylation status at three HpaII-cutting CpG sites of the H19 promoter in adrenocortical carcinomas and in a virilizing adenoma than in their adjacent normal adrenal tissues. Bisulfite genomic sequencing revealed a significantly higher mean degree of methylation at each of 12 CpG sites of the H19 promoter in adrenocortical carcinomas than in normal adrenals (P < 0.01 for all sites) or adrenocortical adenomas (P < 0.01, except P < 0.05 for site 12 and P > 0.05 for site 11). The mean methylation degree of the 12 CpG sites was significantly higher in the adrenocortical carcinomas (mean +/- SE, 76 +/- 7%) than in normal adrenals (41 +/- 2%) or adrenocortical adenomas (45 +/- 3%; both P < 0.005). RNA analysis indicated that the adrenocortical carcinomas expressed less H19 but more IGF-II RNAs than normal adrenal tissues did. The mean methylation degree of the 12 H19 promoter CpG sites correlated negatively with H19 RNA levels (r = -0.550; P < 0.01), but positively with IGF-II mRNA levels (r = 0.805; P < 0.001). In the adrenocortical carcinoma cell line NCI-H295R, abundant IGF-II, but minimal H19, RNA expression was detected by Northern blotting. Treatment with a cytosine methylation inhibitor, 5-aza-2' deoxycytidine, increased H19 RNA expression, whereas it decreased IGF-II mRNA accumulation dose- and time-dependently (both P < 0.005) and reduced cell proliferation to 10% in 7 d. Our results suggest that altered DNA methylation of the H19 promoter is involved in the abnormal expression of both H19 and IGF-II genes in human adrenocortical carcinomas. PMID- 11889183 TI - Glutamic acid decarboxylase antibody positivity is associated with an impaired insulin response to glucose and arginine in nondiabetic patients with autoimmune thyroiditis. AB - To study whether antibodies to glutamic acid decarboxylase (GADab) are associated with subclinical beta-cell damage and impaired insulin secretion, we screened 441 nondiabetic patients with autoimmune thyroiditis (AT) for GADab, and 15 (3.4%) were found positive. Antibodies to IA-2 were found in two GADab+ and one GADab- patients. We matched 11 GADab+ and 13 GADab- AT patients who were euthyroid on thyroxin supplementation, and 13 control subjects for sex, age, and body mass index and measured insulin, C-peptide, and glucagon response to glucose and arginine at three blood glucose concentrations (fasting, 14 mmol/liter, >25 mmol/liter). In the fasting state, all groups had similar blood glucose concentration and HbA1c level, but the serum insulin concentration was higher in the AT patients compared with the control subjects (P < 0.04). The acute insulin response to arginine was lower in GADab+ than in GADab- thyroiditis subjects at glucose concentration of 14 and >25 mmol/liter (AIR(14): 76.8 +/- 52.0 vs. 158.2 +/- 118.2 mU/liter, P = 0.040; AIR(>25): 84.3 +/- 64.4 vs. 167.9 +/- 101.5 mU/liter, P = 0.035). In conclusion, GADab were associated with a decreased insulin secretion capacity in nondiabetic subjects with thyroiditis, which suggests that GADab positivity could be a marker of subclinical insulitis. PMID- 11889184 TI - IL-1 receptor antagonist serum levels are increased in human obesity: a possible link to the resistance to leptin? AB - We have recently shown that human monocytic cells express functional leptin receptors and that leptin is capable of inducing the expression and secretion of the IL-1 receptor antagonist (IL-1Ra). Although IL-1Ra has anti-inflammatory and possibly anti-atherogenic properties, it has also been shown to antagonize the action of leptin at the hypothalamic level in rodents, thereby inducing leptin resistance. We have therefore examined whether IL-1Ra levels are increased in human hyperleptinemic conditions, such as obesity. To this end, we measured serum IL-1Ra levels in 20 morbidly obese nondiabetic subjects [body mass index (BMI), 45 +/- 6 kg/m(2); serum leptin, 52 +/- 20 ng/ml] as well as in 10 age- and sex matched lean controls (BMI, 22 +/- 2 kg/m(2); serum leptin, 7 +/- 4 ng/ml). Serum IL-1Ra concentrations proved to be elevated 6.5-fold in the obese subjects, and they were positively correlated in a linear manner with the leptin levels (r(2) = 0.34; P = 0.01), although lean body mass (LBM) and the insulin resistance index were even better predictors of IL-1Ra levels (r(2) = 0.45 and 0.58, respectively; P < 0.01). Six months after 15 of the 20 obese subjects had undergone bypass surgery for their morbid obesity, their mean BMI and leptin levels decreased to 33 +/- 7 kg/m(2) and 18 plus minus 12 ng/ml, respectively. This change in leptin concentrations was associated with a significant reduction in IL-1Ra levels (P < 0.02). However, there was a better correlation between the decrease in IL-1Ra level and the change in LBM than with the reduction in leptin levels, indicating that leptin is not the sole determinant of circulating IL-1Ra in obesity. In summary, we demonstrate that IL-1Ra levels are highly elevated in human obesity and that its concentrations decrease after weight loss from bypass surgery. However, LBM and insulin resistance are better predictors of serum IL-1Ra concentrations than are leptin levels, suggesting that additional metabolic factors control the secretion of this cytokine antagonist. Although the immunological consequences of this alteration remain unknown, it is tempting to speculate that the obesity-related increase in IL-1Ra might contribute to the central resistance to leptin in obese patients, similar to the inhibition of the hypothalamic signaling of leptin by IL-1Ra in rodents. PMID- 11889186 TI - Ontogeny of human fetal testicular apoptosis during first, second, and third trimesters of pregnancy. AB - During spermatogenesis in human adults, testicular germ cells proliferate, differentiate, and die by apoptosis. However, little is known about the temporal or spatial nature of this programmed cell death. Such information may be useful for understanding prenatal developmental biology as well as spermatogenesis during adulthood, particularly in the context of germ cell disorders. We undertook this study to determine 1) whether apoptosis occurred in a cell specific fashion in the germ cell population and the supporting somatic cells; and 2) whether apoptosis varied with gestational age. We examined human fetal testicular tissues obtained from 17 karyotypically and structurally normal fetuses of mothers who underwent spontaneous or induced abortions. Three gestational ages were defined as follows: group A, 12-13 wk gestation (n = 5); group B, 20-22 wk gestation (n = 7); and group C, 37-40 wk gestation (n = 5). Morphology in conjunction with in situ end labeling was used to identify and quantify apoptotic nuclei in fetal gonadal tissues. The results of this study suggest that gonadal apoptosis occurred in germ cells, Sertoli cells, and Leydig cells at all gestational ages. Apoptotic death was highest in the Leydig cells, followed by germ cells and Sertoli cells. There was a significant positive correlation between the apoptosis of germ cells and Sertoli cells (P < 0.01) and a negative correlation between healthy germ cells and Sertoli cells (P < 0.001). There was also a negative correlation between the intratubular cell number and the gestational age. Specifically, the proportion of Sertoli cells decreased with gestational age, although there was no significant change in the germ cell in relation to gestational age. No such relationship was found in the Leydig cell population, all of which reside outside the seminiferous tubules. These results are the first to suggest that fetal testicular apoptosis begins in the first trimester, occurs in the three major cell types, and continues throughout pregnancy. Our data also suggest that in the fetal gonad, germ and Sertoli cell proliferation and death may be controlled by a genetic program distinct from that of the Leydig cells. This information is relevant to the understanding of abnormal spermatogenesis associated with infertility and to germ cell tumors in adult life. PMID- 11889187 TI - Fetal growth and the function of the adrenal cortex in preterm infants. AB - Small for gestational age preterm infants have a higher risk of neonatal morbidity compared to appropriate for gestational age preterm infants. A diminished adrenal response to stress may be involved in the higher postnatal morbidity. The adrenal cortex response in relation to fetal growth was studied by ACTH stimulation tests in 43 preterm infants (born < or = 32 wk). The cortisol and 17-hydroxyprogesterone (17-OHP) responses to 1 microg/kg ACTH were analyzed in relation to birth weight SD scores (BW-SDS) corrected for gestational age, gender, and parity. BW-SDS was significantly associated with the cortisol and 17 OHP response. Infants with the lowest BW-SDS had the lowest cortisol levels after stimulation. No effect of size at birth was found on the ratio between cortisol and 17-OHP. In addition, basal cortisone levels in a single blood sample were higher in infants with the lowest BW-SDS than in infants with higher BW-SDS, but the ratio between cortisol and cortisone was comparable in the two groups. We conclude that the response of cortisol and 17-OHP to ACTH stimulation in preterm infants is related to fetal growth. The lack of influence of fetal growth on the ratio between cortisol and 17-OHP after ACTH stimulation suggests that the activities of 21- and 11 beta-hydroxylase are not affected. The lower adrenal response to stimulation may be important in neonatal morbidity and possibly the development of disease in later life in growth-restricted preterm infants. PMID- 11889188 TI - Cortisol and 17-hydroxyprogesterone kinetics in saliva after oral administration of hydrocortisone in children and young adolescents with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - We have analyzed the kinetics of salivary cortisol (F) and 17-hydoxyprogesterone (17OHP) after a single oral administration of hydrocortisone (HC; 10 mg; 0700 h) in healthy male volunteers (n = 10; 18-29 yr) and in patients with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (males, n = 7; females, n = 3; 8.5-20.4 yr). The HC doses, related to body surface area, ranged from 6.3-9.2 mg/m(2) in controls and from 4.2-10.7 mg/m(2) in CAH patients. Saliva was collected over 5 h (at intervals of 15-30 min), and the steroids were measured with adapted RIAs. In healthy controls, maximal cortisol values (250.3 +/- 35.9 nmol/liter) were reached after 30 min. Values showed a monophasic decrease. A t(1/2) of 94.5 min was calculated. The proportion of the HC dose in the total area under the curve was 71.2 +/- 3.2%. For 17OHP, a monophasic decrease was found, with a minimum level of 48 +/- 27 pmol/liter after 300 min. In CAH patients the salivary steroid profiles showed individual kinetics (maximal cortisol values ranged from 107-726 nmol/liter). Here a monophasic decrease was found with a shorter t(1/2) of 56.4 min. The HC dose proportion in the area under the curve was 88.3 +/- 6%. 17OHP showed biphasic courses with a decrease to the minimum 17OHP level after 210 min at the latest and a subsequent gradual increase. Our findings of limited normalization of the adrenal cortex by oral HC administration underlines the necessity of optimizing therapy control and indicates the usefulness of kinetic studies for the judgement of therapy in CAH patients. PMID- 11889189 TI - A switch in dehydrogenase to reductase activity of 11 beta-hydroxysteroid dehydrogenase type 1 upon differentiation of human omental adipose stromal cells. AB - As exemplified in patients with Cushing's syndrome, glucocorticoids play an important role in regulating adipose tissue distribution and function, but circulating cortisol concentrations are normal in most patients with obesity. However, human omental adipose stromal cells (ASCs) can generate glucocorticoid locally through the expression of the enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) type 1 (11 beta-HSD1), which, in intact cells, has been considered to be an oxoreductase, converting inactive cortisone (E) to cortisol (F). Locally produced F can induce ASC differentiation, but the relationship between 11 beta HSD1 expression and adipocyte differentiation is unknown. Primary cultures of paired omental (om) and sc ASC and adipocytes were prepared from 17 patients undergoing elective abdominal surgery and cultured for up to 14 d. Expression and activity of 11 beta-HSD isozymes were analyzed together with early (lipoprotein lipase) and terminal (glycerol 3 phosphate dehydrogenase) markers of adipocyte differentiation. On d 1 of culture, 11 beta-HSD1 activity in intact om ASCs exceeded oxoreductase activity in every patient (78.9 +/- 24.9 vs. 15.8 +/- 3.7 [mean +/- SE] pmol/mg per hour, P < 0.001), and in sc ASCs, relative activities were similar (40.6 +/- 12.2 vs. 36.9 +/- 8.8). Conversely, in freshly isolated om adipocytes, reductase activity exceeded dehydrogenase activity (23.6 +/- 1.5 vs. 6.2 +/- 0.8 pmol/mg per hour, P < 0.01). Following 14 d of culture in serum-free conditions with addition of 10 nM insulin (Ctr) or insulin with 100 nM F (+F), lipoprotein lipase/18S RNA levels increased in both the Ctr- and +F-treated ASCs, but glycerol 3 phosphate dehydrogenase increased only in the +F cultures. In both cases, however, 11 beta-HSD1 oxoreductase activity exceeded dehydrogenase activity (Ctr: 53.3 +/- 9.0 vs. 32.4 +/- 10.5, P < 0.05; +F: 65.6 +/- 15.6 vs. 37.1 +/- 11.5 pmol/mg per hour, P < 0.05), despite no significant changes in 11 beta-HSD1 mRNA levels. In sc ASCs, dehydrogenase activity was similar to reductase activity in both Ctr- and +F-treated cells. Type 2 11 beta-HSD expression was undetectable in each case. These data show that in intact, undifferentiated om ASCs, 11 beta-HSD1 acts primarily as a dehydrogenase, but in mature adipocytes oxoreductase activity predominates. Because glucocorticoids inhibit cell proliferation, we postulate that 11 beta-HSD1 activity in uncommitted ASCs may facilitate proliferation rather than differentiation. Once early differentiation is initiated, a "switch" to 11 beta-HSD1 oxoreductase activity generates F, thus promoting adipogenesis. Site-specific regulation of the set-point of 11 beta-HSD1 activity may be an important mechanism underpinning visceral obesity. PMID- 11889191 TI - Characterization of T(4)-binding globulin cleaved by human leukocyte elastase. AB - T(4)-binding globulin (TBG) serves to maintain an important serum pool of thyroid hormones and to prevent their excessive loss in urine. TBG has also been implicated in the tissue distribution and targeted delivery of the hormones, the mechanisms of which remain unclear. By virtue of sequence homology, TBG belongs to the serine proteinase inhibitors superfamily of proteins that are characterized by a reactive site loop serving as a recognition site for serine proteinases. However, both TBG and another serpin with hormone transport function, corticosteroid-binding globulin, are noninhibitory. Cleavage of corticosteroid-binding globulin by human leukocyte elastase results in the reduction of its hormone-binding affinity and capacity. In this communication we confirm previous observations that TBG is also cleaved by elastase and undergoes the characteristic conformational changes. In addition, contrary to a previous report, the present work demonstrates that the cleaved product has reduced T(4) binding affinity and, as expected, increased heat stability. Additional fragmentation of the molecule results in the loss of the hormone-binding site that is in agreement with a recent in vivo observation of apparent consumption at sites of inflammation. These data suggest that TBG may play a role in the targeted delivery of thyroid hormones to tissues rich in proteinases. PMID- 11889190 TI - Characterization of serotonin(4) receptors in adrenocortical aldosterone producing adenomas: in vivo and in vitro studies. AB - We have previously shown that serotonin (5-HT) stimulates aldosterone secretion from the human adrenal gland through activation of 5-HT(4) receptors. The aim of the present study was to investigate in vivo and in vitro the presence of 5-HT(4) receptors in aldosterone-producing adenomas (aldosteronomas). Eight patients with aldosteronoma received a single oral dose of placebo or cisapride (10 mg). Cisapride administration significantly increased plasma aldosterone within 120 min without any significant change in renin, cortisol, or potassium levels. In two patients, a marked decrease in the plasma aldosterone response to cisapride was observed after surgical removal of the tumor. The effects of 5-HT and selective 5-HT(4) ligands on aldosterone production from aldosteronoma tissues were studied in vitro using a perifusion system technique. 5-HT and the 5-HT(4) receptor agonist cisapride (10(-7) M, 20 min) both stimulated aldosterone secretion from aldosteronoma slices. The 5-HT- and cisapride-evoked aldosterone responses were inhibited by concomitant administration of the specific 5-HT(4) receptor antagonist GR 113808 (10(-7) M, 150 min). PCR amplification revealed the expression of 5-HT(4) receptor mRNA in 13 of 14 aldosteronomas studied. Taken together, these data show that most aldosteronomas, like normal glomerulosa cells, express a functional 5-HT(4) receptor. Our results also suggest that 5-HT, which can be locally released by intratumoral mast cells, may play a role in the pathophysiology of these tumors. PMID- 11889192 TI - Resveratrol induces apoptosis in thyroid cancer cell lines via a MAPK- and p53 dependent mechanism. AB - Two papillary thyroid carcinoma (PTC) and two follicular thyroid carcinoma (FTC) cell lines treated with resveratrol (RV), 1-10 microM, showed activation and nuclear translocation of MAPK (extracellular signal-regulated kinase 1/2). Cellular abundance of the oncogene suppressor protein p53, serine phosphorylation of p53, and abundance of c-fos, c-jun, and p21 mRNAs were also increased by RV. Inhibition of the MAPK pathway by either H-ras antisense transfection or PD 98059, an MAPK kinase inhibitor, blocked these RV-induced effects. Addition of pifithrin-alpha, a specific inhibitor of p53, or transfection of p53 antisense oligonucleotides caused decreased RV-induced p53 and p21 expression in PTC and FTC cells. Studies of nucleosome levels estimated by ELISA and of DNA fragmentation showed that RV induced apoptosis in both papillary and follicular thyroid cancer cell lines; these effects were inhibited by pifithrin-alpha and by p53 antisense oligonucleotide transfection. PD 98059 and H-ras antisense transfection also blocked induction of apoptosis by RV. Thus, RV acts via a Ras MAPK kinase-MAPK signal transduction pathway to increase p53 expression, serine phosphorylation of p53, and p53-dependent apoptosis in PTC and FTC cell lines. PMID- 11889193 TI - Glucose tolerance during moderate alcohol intake: insights on insulin action from glucose/lactate dynamics. AB - Moderate alcohol (ETOH) intake has been associated with a significant reduction in risk for infarction among general populations. In this study, we assessed the effects of low-dose ETOH (40 g over 3-h period as vodka) on the interaction between glucose (G), insulin, and lactate (L) during the insulin-modified frequently sampled i.v. glucose tolerance test (FSIGT) (0.3 U/kg body weight between 20-25 min) in eight normal volunteers. In the control (C) study, water was administered. An insulin-independent two-compartment model was used to describe G and L kinetics. Insulin sensitivity (S(I)) was significantly higher in the ETOH study than in the C study (2.49 +/- 0.52 vs. 0.92 +/- 0.20 10(-4) min( 1)microU(-1)ml, C vs. ETOH; P = 0.0391). No significant differences were observed in G effectiveness (0.029 +/- 0.004 vs. 0.033 +/- 0.004 min(-1)). Blood L levels were higher during FSIGT when ETOH was administered [area under the curve (AUC), 201 plus minus 16 vs. 123 +/- 23 mmol/liter in 240 min; P = 0.0001]. The dynamic analysis of blood L concentrations showed that ETOH also significantly decreased L clearance (0.0016 +/- 0.0011 vs. 0.0029 +/- 0.0002 min(-1); P = 0.0156), whereas no difference was observed for the fractional conversion of the rate of G disappearance to L (0.0033 +/- 0.0012 vs. 0.0031 +/- 0.0005 min(-1)). ETOH decreased baseline plasma FFA concentration; AUC of FFA was markedly reduced with ETOH (65 +/- 14 vs. 109 plus minus 17 mmol/liter in 240 min; P = 0.0063) and inversely correlated with S(I) (r = 0.693; P = 0.0029). The amount of C-peptide in 240 min as well as the amounts before and after insulin administration were not different between the two tests. We concluded that G and L kinetics derived from FSIGT shows that moderate ETOH intake: 1) improves insulin action; 2) decreases L clearance; and 3) does not affect beta cell function. Because ETOH at moderate doses has a marked antilipolytic action, it might improve insulin action by improving substrate competition. The present findings suggest that moderate alcohol consumption in the diet should not be discouraged. PMID- 11889194 TI - Effects of glucagon-like peptide 1 on counterregulatory hormone responses, cognitive functions, and insulin secretion during hyperinsulinemic, stepped hypoglycemic clamp experiments in healthy volunteers. AB - Glucagon-like peptide 1 (GLP-1) and analogues are being evaluated as a new therapeutic principle for the treatment of type 2 diabetes. GLP-1 suppresses glucagon secretion, which could lead to disturbances of hypoglycemia counterregulation. This has, however, not been tested. Nine healthy volunteers with normal oral glucose tolerance received infusions of regular insulin (1 mU x kg(-1) x min(-1)) over 360 min on two occasions in the fasting state. Capillary glucose concentrations were clamped at plateaus of 4.3, 3.7, 3.0, and 2.3 mmol/liter for 90 min each (stepwise hypoglycemic clamp); on one occasion, GLP-1 (1.2 pmol x kg(-1) x min(-1)) was administered i.v. (steady-state concentration, approximately 125 pmol/liter); on the other occasion, NaCl was administered as placebo. Glucagon, cortisol, GH (immunoassays), and catecholamines (radioenzymatic assay) were determined, autonomous and neuroglucopenic symptoms were assessed, and cognitive function was tested at each plateau. Insulin secretion rates were estimated by deconvolution (two-compartment model of C peptide kinetics). At insulin concentrations of approximately 45 mU/liter, glucose infusion rates were similar with and without GLP-1 (P = 0.26). Only during the euglycemic plateau (4.3 mmol/liter), GLP-1 suppressed glucagon concentrations (4.1 +/- 0.4 vs. 6.5 +/- 0.7 pmol/liter; P = 0.012); at all hypoglycemic plateaus, glucagon increased similarly with GLP-1 or placebo, to maximum values greater than 20 pmol/liter (P = 0.97). The other counterregulatory hormones and autonomic or neuroglucopenic symptom scores increased, and cognitive functions decreased with decreasing glucose concentrations, but there were no significant differences comparing experiments with GLP-1 or placebo, except for a significant reduction of GH responses during hypoglycemia with GLP-1 (P = 0.04). GLP-1 stimulated insulin secretion only at plasma glucose concentrations of at least 4.3 mmol/liter. In conclusion, the suppression of glucagon by GLP-1 does occur at euglycemia, but not at hypoglycemic plasma glucose concentrations (< or = 3.7 mmol/liter). GLP-1 does not impair overall hypoglycemia counterregulation except for a reduction in GH responses, which is in line with other findings demonstrating pituitary actions of GLP-1. Below plasma glucose concentrations of 4.3 mmol/liter, the insulinotropic action of GLP-1 is negligible. PMID- 11889195 TI - Iodide kinetics and experimental (131)I therapy in a xenotransplanted human sodium-iodide symporter-transfected human follicular thyroid carcinoma cell line. AB - Uptake of iodide is a prerequisite for radioiodide therapy in thyroid cancer. However, loss of iodide uptake is frequently observed in metastasized thyroid cancer, which may be explained by diminished expression of the human sodium iodide symporter (hNIS). We studied whether transfection of hNIS into the hNIS deficient follicular thyroid carcinoma cell line FTC133 restores the in vivo iodide accumulation in xenografted tumors and their susceptibility to radioiodide therapy. In addition, the effects of low-iodide diets and thyroid ablation on iodide kinetics were investigated. Tumors were established in nude mice injected with the hNIS-transfected cell line FTC133-NIS30 and the empty vector transfected cell line FTC133-V4 as a control. Tumors derived from FTC133-NIS30 in mice on a normal diet revealed a high peak iodide accumulation (17.4% of administered activity, measured with an external probe) as compared with FTC133-V4 (4.6%). Half-life in FTC133-NIS30 tumors was 3.8 h. In mice kept on a low-iodide diet, peak activity in FTC133-NIS30 tumors was diminished (8.1%), whereas thyroid iodide accumulation was increased. In thyroid-ablated mice kept on a low-iodide diet, half-life of radioiodide was increased considerably (26.3 h), leading to a much higher area under the time-radioactivity curve than in FTC133-NIS30 tumors in mice on a normal diet without thyroid ablation. Experimental radioiodide therapy with 2 mCi (74 MBq) in thyroid-ablated nude mice, kept on a low-iodide diet, postponed tumor development (4 wk after therapy, one of seven animals revealed tumor vs. five of six animals without therapy). However, 9 wk after therapy, tumors had developed in four of the seven animals. The calculated tumor dose was 32.2 Gy. We conclude that hNIS transfection into a hNIS-defective thyroid carcinoma cell line restores the in vivo iodide accumulation. The unfavorable iodide kinetic characteristics (short half-life) can be partially improved by conventional conditioning with thyroid ablation and low-iodide diet, leading to postponed tumor development after radioiodide therapy. However, to achieve sufficient radioiodide tumor doses for therapy, further strategies are necessary, aiming at the mechanisms of iodide efflux in particular. PMID- 11889196 TI - Activation of the bone morphogenetic protein signaling pathway induces inhibin beta(B)-subunit mRNA and secreted inhibin B levels in cultured human granulosa luteal cells. AB - During the human menstrual cycle the circulating levels of inhibin B, a dimer of inhibin alpha- and beta(B)-subunits, fluctuate in a fashion distinct from that of inhibin A, the alpha-beta(A)-subunit dimer. This suggests that human inhibin subunits are each regulated in a distinct manner in human ovarian granulosa cells by endocrine and local factors. We have previously shown using cultures of human granulosa-luteal (hGL) cells that gonadotropins stimulate the steady state mRNA levels of inhibin alpha- and beta(A)-subunits, but not those of the beta(B) subunit, which, on the other hand, are up-regulated by, for instance, activin and TGF beta. We recently identified the TGF beta gene family member bone morphogenetic protein-3 (BMP-3) as a granulosa cell-derived growth factor, but whether BMP-3 or other structurally related BMPs regulate human granulosa cell inhibin production is not known. We show here that hGL cells express mRNAs for distinct serine/threonine kinase receptors (BMP-RIA and BMP-RII) and Smad signaling proteins (Smad1, Smad4, and Smad5) involved in the mediation of cellular effects of BMPs. Subsequently, we determined in hGL cell cultures the effects of distinct members of the BMP family previously found to be expressed in mammalian ovaries. Recombinant BMP-2 induces potently in a time- and concentration-dependent manner the expression of the inhibin beta(B)-subunit mRNAs in hGL cells without affecting the levels of alpha- or beta(A)-subunit mRNAs. BMP-6 has a similar, but weaker, effect than BMP-2, whereas BMP-3 and its close homolog, BMP-3b (also known as growth differentiation factor-10) had no effect on inhibin subunit mRNA expression. hCG treatment of hGL cells was previously shown to abolish the stimulatory effect of activin on beta(B)-subunit mRNA levels, and here hCG is also shown to suppress the effect of BMP-2. Furthermore, BMP-2 stimulates hGL cell secreted dimeric inhibin B levels in a concentration-dependent manner. Depending on the experiment, maximal increases in inhibin B levels of 6- to 28-fold above basal levels were detected during a 72-h culture period. We conclude that activation of the BMP-signaling pathway in hGL cells stimulates inhibin beta(B)-subunit mRNA levels and leads at the protein level to a dramatic stimulation of secreted inhibin B dimers. Our results are consistent with the suggestion that in addition to the distinct activin- and TGF beta-activated signaling pathways, the BMP-activated pathway is likely to be implicated in the complex regulation of inhibins in the human ovary. PMID- 11889197 TI - Loss of expression of GADD45 gamma, a growth inhibitory gene, in human pituitary adenomas: implications for tumorigenesis. AB - The underlying molecular pathogenic mechanisms remain unknown in the majority of human pituitary tumors. GADD45 gamma is a member of a growth arrest and DNA damage-inducible gene family that functions in the negative regulation of cell growth. We have found that the mRNA expression of the GADD45 gamma gene is significantly different between normal human pituitary tissue and clinically nonfunctioning pituitary adenomas using cDNA-representational difference analysis. Although GADD45 gamma mRNA was found in normal human pituitary tissue, it was detectable in only 1 of 18 clinically nonfunctioning pituitary tumors by RT-PCR. Furthermore, this gene was not expressed in the majority of GH- or PRL secreting pituitary tumors (6 of 8 and 7 of 10, respectively). In colony formation assays, transfection of human GADD45 gamma cDNA into the human pituitary tumor-derived cell line, PDFS, results in a dramatic decrease in cell growth by 88%. GADD45 gamma also reduces colony formation in other pituitary tumor-derived cell lines, AtT20 and GH4, by approximately 60% and 50%, respectively, confirming its function in controlling cell proliferation in the pituitary. These data indicate that GADD45 gamma is a powerful growth suppressor controlling pituitary cell proliferation, and GADD45 gamma represents the first identified gene whose expression is lost in the majority of human pituitary tumors. PMID- 11889198 TI - Paraoxonase-1 L55M polymorphism is associated with an abnormal oral glucose tolerance test and differentiates high risk coronary disease families. AB - Polymorphisms of the gene for the antioxidant enzyme, paraoxonase-1 (PON1), have been identified as risk factors for coronary disease (CHD), notably in diabetic patients. The polymorphisms have also been linked with other diabetic complications. The present study analyzed glucose metabolism as a function of PON1 polymorphisms in young healthy nondiabetic men from families with premature CHD and matched controls. The L55M PON1 polymorphism was independently associated with the glucose response to an oral glucose tolerance test. LL homozygotes had significantly impaired glucose disposal (P = 0.0007) compared with (LM+MM) genotypes. It was particularly marked for subjects from high CHD risk families and differentiated them from matched controls (P = 0.049). The area under the glucose curve (P = 0.0036) and the time to peak glucose value (P = 0.026) were significantly higher in the LL carriers, whereas the insulin response was slower (P = 0.013). Insulin resistance did not differ between L55M genotypes. There was a trend for reduced pancreatic beta-cell function as measured by glucose-induced insulin secretion (LL vs. LM vs. MM, 20.26 vs. 23.74 vs. 25.60; P = 0.077). The frequency of the L55 allele decreased significantly (P = 0.028) across regions defining a north-south European axis. No significant differences for the glucose response or case-control populations were observed as a function of the PON1 Q192R polymorphism. The study demonstrates an association between PON1 gene polymorphisms and glucose metabolism. The L55M-glucose interaction differentiated offspring of high CHD risk families, suggesting that it may be of particular relevance for vascular disease and possibly other diabetic complications. PMID- 11889199 TI - Hypocaloric diet reduces exercise-induced alpha 2-adrenergic antilipolytic effect and alpha 2-adrenergic receptor mRNA levels in adipose tissue of obese women. AB - Previous investigations have shown that alpha 2-adrenoceptor (alpha 2-AR) stimulation blunts lipid mobilization during physiological activation of the sympathetic nervous system promoted by exercise in sc abdominal adipose tissue (SCAAT) in obese men. To investigate the effect of a low calorie diet (LCD) on the alpha 2-adrenergic responsiveness and on the expression of alpha 2-AR and beta 2-adrenoceptor (beta 2-AR) in SCAAT, 11 obese women (weight: 99.1 +/- 4.6 kg; body mass index: 34.3 +/- 1.1 kg/m(2)) received a 12-wk diet providing 500 kcal/d less than their usual diet. The exercise-induced alpha 2-adrenergic antilipolytic effect was investigated in SCAAT before and at the end of LCD. Changes in extracellular glycerol concentration and local blood flow were measured in SCAAT during a 45-min exercise bout (50% of heart rate reserve) using a control microdialysis probe and a probe supplemented with the alpha2-AR antagonist phentolamine. SCAAT biopsies were performed for determination of mRNA levels using RT-competitive PCR. Plasma catecholamine responses to exercise bout were not different before and at the end of LCD. Before LCD, the exercise-induced increase in extracellular glycerol concentration was potentiated by phentolamine supplementation, while this potentiating effect of the alpha-antagonist was not observed at the end of LCD. No changes were observed for beta 2-AR and hormone sensitive lipase mRNA levels, while alpha 2-AR mRNA level was significantly decreased in adipose tissue during LCD. These findings show that alpha 2-AR mediated antilipolytic action is reduced by a moderate hypocaloric diet and that down-regulation of alpha 2-AR mRNA levels may participate in the decrease of the alpha 2-adrenergic effect revealed by microdialysis. PMID- 11889200 TI - The insulinotropic effect of acute exendin-4 administered to humans: comparison of nondiabetic state to type 2 diabetes. AB - Exendin-4 is a potent and long-acting agonist of the glucagon-like peptide-1 (GLP 1) receptor. GLP-1 is an insulinotropic gut peptide and is being evaluated for the regulation of plasma glucose in type 2 diabetes. The purpose of the present study was to ascertain whether exendin-4 is insulinotropic and whether it has long-lived biological effects in nondiabetic and type 2 diabetic subjects. Because incretins are glucose dependent with respect to their insulin-releasing capacity, we used the hyperglycemic glucose clamp technique to begin to address these issues in two separate protocols. In one protocol, we infused exendin-4 (0.15 pmol x kg(-1) x min(-1)) in seven nondiabetic and seven type 2 diabetic subjects during the second hour of a 5-h hyperglycemic clamp in which fasting plasma glucose was raised by 5.4 mmol/liter. The second protocol was identical to the first except that plasma glucose was allowed to fall to the fasting levels during the fourth hour and again raised by 5.4 mmol/liter during the fifth hour in four nondiabetic and four diabetic subjects. With the initiation of exendin-4 infusion at 60 min, plasma insulin response was potentiated 4- to 5-fold in both groups. Despite termination of exendin-4 at the end of the second hour, the insulin levels remained elevated for several hours and hyperglycemia was maintained. All volunteers ate a meal 5.5 h after inducing hyperglycemia. Postprandial plasma glucose, insulin, and GLP-1 did not rise in any subject, possibly because of delayed gastric emptying by exendin-4 even though its infusion had been terminated 4 h previously. We concluded that exendin-4 is a potent and long-lasting insulinotropic agent in nondiabetic and diabetic subjects. PMID- 11889201 TI - Structural changes in the angiofollicular units between active and hypofunctioning follicles align with differences in the epithelial expression of newly discovered proteins involved in iodine transport and organification. AB - In animals, as well as in humans, the thyroid gland is made of active follicles, with cuboidal cells and hypofunctioning follicles, with flattened cells. In this study, the functional status of human follicles was dissected out, based on immunohistochemical detection of TSH receptor, Na(+)/I(-) symporter, pendrin, thyroperoxidase (TPO), thyroid oxidases (ThOXs), and T(4)-containing iodinated Tg (Tg-I). To ascertain that angiofollicular units exist in the human, we studied the microvascular bed of each follicle, in correlation with detection of vascular endothelial growth factor (VEGF), of nitric oxide synthase III, and of endothelin in normal and goitrous thyroids. In hypofunctioning follicles, pendrin, TPO, and ThOXs were not detected, and there was no Tg-I in the colloid. At the opposite, in active follicles, pendrin, TPO, and ThOXs were detected in thyrocytes, and Tg I was present in the colloid. In normal and goitrous thyroids, the capillary networks surrounding active follicles were larger than those surrounding hypofunctioning follicles. Immunoreactivity for nitric oxide synthase III and endothelin was solely detected in active follicles. Only a few follicles in normal thyroids were immunostained for VEGF, regardless of their functional status. In multinodular goiters, VEGF was detected in contact with the extracellular matrix at the basal pole of the cells. In conclusion, the present study endorses the likelihood of angiofollicular units in the human thyroids. Vascular changes are related to the functional status of thyrocytes. PMID- 11889202 TI - Expression of ghrelin and of the GH secretagogue receptor by pancreatic islet cells and related endocrine tumors. AB - Ghrelin is a novel gastrointestinal hormone produced by rat and human gastric X like neuroendocrine cells, which strongly stimulates GH secretion and influences energy balance, gastric motility, and acid secretion. Ghrelin is expressed in pituitary and gastrointestinal endocrine tumors. It binds to the GH secretagogue receptor (GHS-R), which is present in a wide variety of central and peripheral human tissues. The aim of the present study was 2-fold: 1) to determine, by immunohistochemistry and mRNA analysis, whether pancreatic islet cells produce ghrelin and express GHS-R; and 2) to investigate ghrelin and GHS-R expression in pancreatic endocrine tumors. Seven cases of nonneoplastic pancreatic tissue and 28 endocrine tumors were studied. In pancreatic islets, ghrelin immunoreactivity was present in all cases and confined to beta-cells. Eleven of the 28 (39%) endocrine tumors were immunoreactive for ghrelin. In situ hybridization and RT PCR confirmed the immunohistochemical data for both tumors and islets but also revealed ghrelin mRNA in 8 and 11 additional tumors, respectively. GHS-R 1a and 1b mRNAs were present in 7 of 28 and 14 of 28 tumors, respectively, studied by RT PCR. These findings demonstrate that ghrelin production is not restricted to the stomach but is also present in pancreatic beta-cells and endocrine tumors (regardless of the type of pancreatic hormone produced, if any). Expression of GHS-R in some of the endocrine tumors studied indicates that autocrine/paracrine circuits may be active in neoplastic conditions. PMID- 11889203 TI - Identification and functional characterization of novel calcium-sensing receptor mutations in familial hypocalciuric hypercalcemia and autosomal dominant hypocalcemia. AB - Familial hypocalciuric hypercalcemia (FHH), neonatal severe hyperparathyroidism (NSHPT), and autosomal dominant hypocalcemia (ADH), in which calcium homeostasis is disordered, are associated with mutations in the calcium-sensing receptor (CASR). Six unrelated kindreds with FHH and/or NSHPT and two unrelated kindreds with ADH were studied. Direct sequence analysis of the exons of the CASR gene identified heterozygous mutations in six of the kindreds with FHH and in one of those with ADH. We performed functional analyses on the novel missense and insertion/frameshift mutants by transiently transfecting wild-type and mutant CASRs tagged with a c-Myc epitope in human embryonic kidney (HEK293) cells. All mutant receptors were expressed at a similar level to that of the wild type; however, whereas mutants R220W and A835T (the ADH mutant) were fully glycosylated and were visualized on the cell surface, glycosylation of mutants G549R and C850 851 ins/fs was impaired, resulting in reduced cell surface staining. In fura-2 loaded HEK293 cells expressing the wild-type or mutant receptors, the inactivating R220W mutant produced a significant shift to the right relative to the wild-type CASR in the cytosolic calcium response to increasing extracellular calcium concentrations and the G549R and C850-851 ins/fs mutants were without detectable activity. The activating A835T mutation resulted in a shift to the left in the cytosolic calcium response to extracellular calcium concentrations relative to the wild type. Our studies have identified novel CASR mutations that alter the function of the CASR in several different ways. PMID- 11889204 TI - Cytochrome P450-catalyzed binding of 3-methylsulfonyl-DDE and o,p'-DDD in human adrenal zona fasciculata/reticularis. AB - 3-Methylsulfonyl-2,2'-bis(4-chlorophenyl)-1,1'-dichloroethene (MeSO(2)-DDE) is a potent, tissue-specific toxicant that induces necrosis of the adrenal zona fasciculata following a local CYP11B1-catalyzed activation to a reactive intermediate in mice. Autoradiography was used to examine CYP11B1-catalyzed binding of MeSO(2)-[(14)C]DDE and the adrenocorticolytic drug 2-(2-chlorophenyl) 2-(4-chlorophenyl)-1,1-dichlorethane; (o,p'-[(14)C]DDD, Mitotane, Lysodren) in human adrenal tissue slice culture. Both compounds gave rise to a selective binding in the one sample of normal adrenal zona fasciculata/reticularis, leaving zona glomerulosa and the adrenal medulla devoid of binding. Addition of the CYP11B1 selective inhibitor metyrapone (50 microM) reduced MeSO(2)-[(14)C]DDE binding below the detection limit, whereas o,p'-[(14)C]DDD binding was reduced only by 42%. Selective binding of MeSO(2)-[(14)C]DDE and o,p'-[(14)C]DDD was also observed in an aldosterone-producing adrenocortical carcinoma and in a nonfunctional adrenocortical hyperplasia. Exposure of slices from the normal adrenal cortex to MeSO(2)-DDE (25 microM) resulted in an increased accumulation of 11-deoxycorticosterone, 11-deoxycortisol and androstenedione in the medium, and exposure to o,p'-DDD (25 microM) did not alter the steroid secretion pattern. No histological changes were found in either MeSO(2)-DDE- or o,p'-DDD-exposed slices, compared with nonexposed slices. We suggest that MeSO(2)-DDE might act as a potent adrenocorticolytic agent in humans. Further studies are needed to establish the usefulness of MeSO(2)-DDE as a possible alternative for the treatment of adrenocortical hypersecretion and tumor growth. PMID- 11889205 TI - Glucocorticoid regulation of p450 aromatase activity in human adipose tissue: gender and site differences. AB - The distinct gender-specific patterns of fat distribution in men and women (android and gynoid) suggest a role for sex steroids. In keeping with these observations, it has been suggested that estrogens can promote preadipocyte cell proliferation and/or differentiation. The enzyme aromatase P450 is responsible for the conversion of androgen precursor steroids to estrogens and may, therefore, have a role in regulating adipose tissue mass and its distribution. We have investigated the glucocorticoid regulation of aromatase expression in human adipose tissue, specifically to define any site- and gender-specific differences. Abdominal subcutaneous (Sc) and omental (Om) adipose tissue was obtained from male and female patients undergoing elective surgery. After collagenase digestion, preadipocytes were cultured in serum-free medium, for 6-10 d, until confluent with either cortisol (10(-6) M, 10(-7) M) or insulin (500 nM) or a combination of both treatments. Adipocytes were studied in suspension cultures. Aromatase activity was assessed using tritiated [1 beta-(3)H]-androstenedione as substrate. In Sc preadipocytes, basal aromatase activity increased in females from 11.5 +/- 1.4 (mean plus minus SEM) to 28.0 +/- 1.8 pmol/mg x h (n = 17, P < 0.05) with 10(-6) M cortisol. By contrast, in males, aromatase activity was inhibited by 10(-6) M cortisol (19.4 +/- 2.4 pmol/mg x h vs. 7.5 +/- 1.3, n = 9, P < 0.01; men vs. women, P < 0.005). These data were endorsed through Western blot analysis using an in-house antihuman aromatase antibody, which recognized a specific 55-kDa species. Aromatase activity was less at Om sites in preadipocytes, increasing in females from 1.1 +/- 0.2 to 3.2 +/- 0.7 pmol/mg x h with 10(-6) M cortisol (P < 0.05) and in males from 2.6 +/- 0.1 pmol/mg x h to 7.8 +/- 0.3 pmol/mg x h after cortisol (men vs. women, P < 0.001). Cortisol induced aromatase activity in Om adipocytes from postmenopausal females was higher than that in premenopausal females (P < 0.001). Insulin had no independent effect on aromatase expression, but coincubation of preadipocytes with cortisol and insulin eliminated both gender- and site-specific differences. In conclusion, in women, but not men, cortisol increased aromatase activity at Sc sites, and this may facilitate predilection for Sc adiposity in females. The observed site-, gender-, and menopausal-specific differences in the glucocorticoid regulation of this enzyme may contribute to the gender- and menopausal-specific patterns of fat distribution. PMID- 11889206 TI - Aberrant expression of growth differentiation factor-9 in oocytes of women with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a major cause of female infertility. Despite substantial effort, the etiology and pathogenesis of PCOS and polycystic ovaries (PCO) in women remain unknown. Recent studies in laboratory animals have documented a link between dysfunction of two oocyte growth factors, growth differentiation factor-9 (GDF-9) and bone morphogenetic factor-15 (BMP-15), and aberrant folliculogenesis. Because aberrant follicle development is a hallmark of PCOS, we wondered whether the expression patterns of these growth factors might be disrupted in PCOS and PCO oocytes. To address this issue, we compared the pattern and level of expression of GDF-9 and BMP-15 mRNA in ovaries from normal cycling (n = 12), PCOS (n = 5), and PCO (n = 7) patients. In situ hybridization studies showed that the expression of GDF-9 and BMP-15 is restricted to the oocytes in all ovaries examined. Interestingly, a decreased level of GDF-9 signal was observed in developing PCOS and PCO oocytes, compared with normal. This difference was evident throughout folliculogenesis, beginning at recruitment initiation and continuing through the small Graafian follicle stage. By contrast, there were no qualitative or quantitative changes in the expression of BMP-15 mRNA in PCOS oocytes during folliculogenesis. There were also no significant differences between normal and PCOS and PCOs in the levels of the mRNA encoding the housekeeping gene, cyclophilin. Together, these results indicate that the expression of GDF-9 mRNA is delayed and reduced in PCOS and PCO oocytes during their growth and differentiation phase. Because oocyte-derived GDF-9 is crucial for normal folliculogenesis and female fertility, we suggest that a dysregulation of oocyte GDF-9 expression may contribute to aberrant folliculogenesis in PCOS and PCO women. PMID- 11889207 TI - Differential changes in 15-hydroxyprostaglandin dehydrogenase and prostaglandin H synthase (types I and II) in human pregnant myometrium. AB - Prostaglandins (PGs) play a key role in the onset of labor in many species and regulate uterine contractility and cervical dilatation. Therefore, the regulation of prostaglandin output by PG synthesizing (PGHS-I and PGHS-II) and metabolizing (PGDH) enzymes in the human myometrium may determine uterine activity patterns in human labor both at preterm and at term. We hypothesized that expression of PGHS isozymes and PGDH in myometrium from women at preterm and term labor would change to favor increased uterotonin (PG) production. Myometrial samples were obtained from the lower uterine segment during cesarean section deliveries from women presenting in preterm, no labor; preterm, labor; term, no labor; term, labor. Immunoreactive (ir-) PGHS and PGDH protein was localized using immunohistochemistry, and changes in protein levels were determined by Western blotting. Ir-PGHS-I and PGHS-II proteins were localized only to myocytes. Ir-PGDH was localized to myocytes in all samples of myometrium examined, but using dual immunofluorescence and immunohistochemistry, ir-PGDH was also detected in cells of the connective tissue. Levels of ir-PGHS-I and PGHS-II protein were not significantly different between no labor and labor tissues, either at preterm or at term. There was no significant effect of gestational age on levels of PGDH, PGHS-I, and PGHS-II protein, but there was a significant decrease in ir-PGDH protein levels in myometrium with labor both at preterm and at term. In addition, there was a decrease in PGDH activity in myometrium from women in labor, both at preterm and at term. Therefore, we conclude that PGDH, PGHS-I, and PGHS-II protein localize within the myocytes of the human pregnant myometrium. A decrease in PGDH protein and activity occurs in association with active labor and may contribute to the amount of bioactive PGs available to act within the human pregnant myometrium at that time. PMID- 11889208 TI - Expression of matrix metalloproteinase (MMP)-2 and MMP-9 in human placenta and fetal membranes in relation to preterm and term labor. AB - Extensive extracellular matrix (ECM) remodeling is found in many processes during human parturition at term and preterm. These include cervical ripening, fetal membrane rupture, and placental detachment from the maternal uterus. Matrix metalloproteinases (MMPs) are the main mediators of ECM degradation. The present study was designed to investigate the expression of MMP-2 and MMP-9 in human fetal membranes (FMs) and placental (PL) tissues with or without labor at preterm and term parturition. Both zymography and Western blot analysis showed that MMP-9 was significantly (P < 0.01) increased in preterm and term labor FM, compared with nonlabor. Term labor PL also had a much higher (P < 0.05) level of MMP-9 than that of term nonlabor. No significant difference in MMP-2 expression was found between labor and nonlabor tissues. Immunolocalization studies revealed a specific distribution pattern for MMP-2 and MMP-9. MMP-2 was localized to the amnion mesenchyme, chorion laeve trophoblast, decidua parietalis, and blood vessels in PL villi. MMP-9 was localized mainly to amnion epithelia, chorion laeve trophoblast, decidua parietalis, and PL syncytiotrophoblasts. Separate cell culture from different layers of FM and culture of purified PL trophoblast cells showed that PL syncytiotrophoblast and amnion epithelial cells exclusively produced MMP-9; chorion trophoblast cells secreted both MMP-2 and MMP-9, but amnion mesenchymal cells produced only MMP-2. We concluded that MMP-2 and MMP-9 exhibited cell-specific expression in the human PL. An increase in MMP-9 expression may contribute to degradation of the ECM in the FM and PL, thereby facilitating FM rupture and PL detachment from the maternal uterus at labor, both preterm and term. PMID- 11889209 TI - Muscarinic receptors in human luteinized granulosa cells: activation blocks gap junctions and induces the transcription factor early growth response factor-1. AB - Acetylcholine (ACh) was recently described to be produced by and act on human luteinizing granulosa cells (GCs). Cholinergic agents increase intracellular calcium levels and stimulate GC proliferation via muscarinic receptors. Based on this observation and because endocrine cells of the forming human corpus luteum (CL), which are the in vivo counterparts of GCs, also proliferate in vivo, we hypothesized that ACh may be a factor involved in the regulation of the complex cellular events occurring during ovulation and formation of the CL. We addressed this possibility by investigating ACh/muscarinic receptor-mediated events in GCs. Normally, cultured GCs and their in vivo counterparts are coupled via gap junctions (GJ) consisting of connexin 43. Treatment with carbachol impaired GJ coupling of GCs within seconds, as shown in single cell, whole cell, patch-clamp studies. The cholinergic antagonist atropine and the muscarinic receptor antagonist pirenzepine specifically blocked this effect. Disruption of GJ communication of GCs is probably due to increased phosphorylation of connexin 43 at serine residues, as shown in immunoprecipitation experiments with carbachol challenged GCs. Ovulation/formation of the CL include reprogramming of luteinizing cells, and in the rat this involves gonadotropin- induced expression of the transcription factor early growth response factor-1 (egr-1). In human GCs we found that carbachol as well as hCG can mimic this effect, as shown by cDNA arrays and semiquantitative RT-PCR. In conclusion, our results obtained in GCs suggest that endogenous, locally produced ACh may contribute to the cellular remodeling of the forming CL via muscarinic receptor/egr-1, thereby affecting proliferation, GJ communication, and regulation of gene expression in luteinizing granulosa cells. PMID- 11889210 TI - In vivo and in vitro evidence suggest that HB-EGF regulates endometrial expression of human decay-accelerating factor. AB - Human endometrium expresses the critical complement component C3 in a cyclic fashion, with the highest expression in the secretory phase. As activated complement can kill cells, self or foreign, the secretory endometrial epithelium protects itself by concomitant expression of complement-protective proteins. The objectives of our present study were to describe the spatial and temporal regulation of the complement-protective protein decay-accelerating factor (DAF) in human endometrium and to identify local regulators of its expression. To describe the cyclic regulation of DAF, immunohistochemistry was performed using the IH4 monoclonal antibody on secretory phase endometrial biopsies taken from normal fertile volunteers in LH-timed cycles (n = 114). DAF expression in human endometrium was predominantly localized to the apical membrane of glandular and luminal epithelium. DAF expression, as assessed by histological scoring analysis, was minimal in the proliferative and early secretory phases and increased markedly on approximately day LH +7 (lumen) and LH +8 (glands). Maximal expression was seen in both glands and lumen by LH +8, and this persisted into menses. Using the RL95-2 endometrial epithelial cancer cell line as a model system, we next examined the cellular regulation of DAF. Treatment with E2 and progesterone, alone or in combination, had little effect on DAF expression. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) treatment increased cell surface and total DAF protein, increasing the signal by 260% on flow cytometry and by 200% on Western blot analysis. Stimulation of DAF protein expression was dose dependent, with maximal expression seen at 1 ng/ml. The stimulatory effects of HB-EGF were also observed at the mRNA level. EGF had effects similar to those of HB-EGF on DAF mRNA and protein expression, suggesting that the HB-EGF effect was mediated at least in part by the Her1 EGF receptor subunit. These studies suggest that DAF expression in the midsecretory phase is stimulated by HB-EGF or other members of the EGF family and may function to protect the epithelial integrity of human endometrium in the face of increased complement expression. PMID- 11889211 TI - Expression and localization of the homeodomain-containing protein HEX in human thyroid tumors. AB - Homeobox genes are involved in neoplastic transformation of both epithelial and hemopoietic tissues. The divergent homeobox gene HEX is expressed in the anterior visceral endoderm during early mouse development and in some adult tissues of endodermal origin, including liver and thyroid. Whereas a role in leukemyogenesis has been proposed already, few data are available on the involvement of HEX in human epithelial tumors. Herein, we analyzed HEX expression and subcellular localization in a series of 55 human thyroid tumors and in several tumoral cell lines. HEX mRNA was detected by RT-PCR either in normal tissues or in thyroid adenomas and differentiated (papillary and follicular) carcinomas. HEX mRNA was also expressed in most undifferentiated carcinomas. Subcellular localization of HEX protein was investigated by immunohistochemistry. In normal tissues and adenomas, HEX protein was present both in nucleus and cytoplasm. In contrast, both differentiated and undifferentiated carcinomas, as well as the tumoral cell lines investigated, showed HEX protein only in the cytoplasm. These findings suggest that regulation of HEX entry in the nucleus of thyrocytes may represent a critical step during human thyroid tumorigenesis. PMID- 11889213 TI - Existence of placental leucine aminopeptidase/oxytocinase/insulin-regulated membrane aminopeptidase in human endometrial epithelial cells. AB - Human endometrial epithelial cells are known to express oxytocin receptors around the time of ovulation. Moreover, oxytocin (OT) and OT-induced prostaglandins appear to play a pivotal role in the switching of endometrial glands from the proliferative to the secretory phase. However, there have been few studies of oxytocinase (OTase), which is identical to placental leucine aminopeptidase (P LAP)/insulin-regulated membrane aminopeptidase (IRAP). We confirmed the expression of P-LAP/OTase in human endometrium and also observed the changes in the expression of P-LAP/OTase throughout the menstrual cycle. P-LAP/OTase and its mRNA were localized in endometrial epithelial cells but not in stromal cells. In the follicular phase, immunoreactive P-LAP/OTase was homogeneously distributed on the plasma membrane and in cytoplasmic granules. Immunoblot analysis demonstrated that the majority of P-LAP/OTase was produced around the time of ovulation. After ovulation, the immunostaining was restricted to the glycogen-rich subnuclear vacuoles, a glandular marker of progesterone release from the corpus luteum. Thereafter, the membrane-bound P-LAP/OTase was released by apocrine secretion during the period of blastocyst implantation and became depleted toward the time of menstruation. Further understanding of the function of P-LAP/OTase in the endometrium appears likely to yield insights into the cyclic changes during the normal menstrual cycle. PMID- 11889214 TI - SHOX nullizygosity and haploinsufficiency in a Japanese family: implication for the development of Turner skeletal features. AB - We report on clinical and molecular findings in a Japanese family consisting of a male infant with SHOX nullizygosity and his four family members with SHOX haploinsufficiency. The male infant had Langer mesomelic dysplasia, the prepubertal sister had idiopathic short stature phenotype with no discernible skeletal features, the father had mild Leri-Weill dyschondrosteosis (LWDC), and the mother and the maternal grandmother had moderate LWDC. The five subjects lacked clinically recognizable short metacarpals, cubitus valgus, high arched palate, short neck, and micrognathia, as well as recurrent otitis media and hearing loss. Fluorescence in situ hybridization and sequence analyses showed that the proband had a pseudoautosomal microdeletion involving SHOX and a C502T missense mutation in the homeobox domain at exon 4, and that the father was heterozygous for the SHOX deletion, and the sister, the mother, and the grandmother were heterozygous for the C502T mutation. The results, in conjunction with the previous findings, suggest that mesomelic skeletal features such as Langer mesomelic dysplasia and LWDC, which are absent or rare in Turner syndrome, are primarily caused by the SHOX dosage effect and the bone maturing effect of gonadal estrogens, whereas other skeletal features such as short metacarpals, cubitus valgus, and various craniofacial and cervical skeletal stigmata, which are common in Turner syndrome, are largely contributed by a compressive effect of distended lymphatics and lymphedema on the developing skeletal tissues. PMID- 11889215 TI - Inhibin-activin receptor subunit gene expression in ovarian tumors. AB - Granulosa cell tumors of the ovary (GCT) express the inhibin subunit genes and secrete dimeric inhibin. Transgenic mice null for the alpha-inhibin gene develop GCT. It has been suggested that this apparent contradiction may be reconciled if the human GCT are resistant to the tumor-suppressive effects of inhibin. Inhibin receptors have recently been characterized as consisting of either betaglycan or p120 in association with the type II or type I activin receptor subunits (ActR), respectively. To test the hypothesis that GCT may exhibit loss of inhibin receptor expression we have examined the expression of the receptor subunits in a cohort of GCT and in mucinous and serous cystadenocarcinomas and normal ovary. Expression was determined by RT-PCR using gene-specific primers and probes combined with Southern blot analysis of the PCR products. The ActRI subunits and ActRIIA exhibited widespread albeit variable expression across tissues, with the highest levels in the serous tumors. ActRIIB expression was relatively low in the mucinous tumors and high in the GCT. Betaglycan expression was abundant, widespread, and variable across all tissues; highest mean levels occurred in the GCT and normal ovary. p120 expression was low or absent in all tissues except the GCT. Within the GCT there was parallel expression of the ActR subunits, betaglycan and p120; the levels, however, varied considerably between tumors. Expression of betaglycan and p120 in most GCT argues against the hypothesis, but does not exclude the possibility that low or absent expression of p120 might be significant in a subset of these tumors. PMID- 11889216 TI - Deletions of the homeobox gene SHOX (short stature homeobox) are an important cause of growth failure in children with short stature. AB - Short stature, with an incidence of 3 in 100, is a fairly frequent disorder in children. Idiopathic short stature refers to patients who are short due to various unknown reasons. Mutations of a human homeobox gene, SHOX (short stature homeobox), have recently been shown to be associated with the short stature phenotype in patients with Turner syndrome and most patients with Leri-Weill dyschondrosteosis. This study addresses the question of the incidence and type of SHOX mutations in patients with short stature. We analyzed the SHOX gene for intragenic mutations by single strand conformation polymorphism, followed by sequencing, in 750 patients and for complete gene deletions by fluorescence in situ hybridization in 150 patients (total, 900 patients). This is the largest group of patients with short stature studied to date for SHOX mutations. All patients had a normal karyotype, and their height for chronological age were below the third percentile or minus 2 SD of national height standards. All were without obvious skeletal features reminiscent of the Leri-Weill syndrome at the time of diagnosis. Silent, missense, and nonsense mutations and a small deletion in the coding region of SHOX were identified in 9 of the 750 patients analyzed for intragenic mutations. Complete gene deletions were detected in 3 of the 150 patients studied for gene deletions. At least 3 of the 9 intragenic mutations were judged to be functional based upon the genotype- phenotype relationship for the parents and normal control individuals. We conclude that SHOX mutations have been detected in 2.4% of children with short stature. The spectrum of SHOX mutations is biased, with the vast majority leading to complete gene deletions. The prevalence of short stature due to SHOX gene mutations among children with short stature appears to be similar to that of GH deficiency or Turner syndrome. Family studies of the children with SHOX mutations often reveal older family members with same mutation who exhibit mild skeletal features reminiscent of the Turner syndrome, such as high-arched palate, short neck, abnormal auricular development, cubitus valgus, genu valgum, short fourth metacarpals, and Madelung deformity. PMID- 11889217 TI - Cell-specific viral gene therapy of a Hurthle cell tumor. AB - We evaluated the effectiveness of a replication-defective adenovirus-transducing thymidine kinase (TK) gene under the control of the rat Tg (rTg) promoter (AdrTgtk) in therapy of a human Hurthle cancer (XTC-1 cell) in vitro and in vivo. The ganciclovir (GCV) sensitivity of infected XTC-1 cells was assessed in vitro by H(3)-thymidine incorporation assay and Trypan-blue exclusion, and by an in vivo tumor development assay. Proliferation was strongly inhibited by adding GCV into the culture medium of infected cells, but not uninfected cells, proving cell infection and expression of TK in the XTC-1 cells. AdrTgtk, and also viruses that have the noncell-specific cytomegalovirus (CMV) promoter-directing expression of TK (AdCMVtk), or luciferase (AdCMVLuc), were used to transduce XTC-1 cells to evaluate killing effects. After infection with AdCMVtk or AdrTgtk, followed by GCV treatment, 70% of infected cells were killed in the presence of GCV, compared with less than 20% of cells infected by AdCMVLuc and treated with GCV. In vivo toxicity was studied in BALB/c mice. When adenovirus is given iv, liver is the major organ infected. No significant changes of the serum transaminase levels and no histological abnormalities were found in animals treated with AdrTgtk/GCV given iv, compared with control animals. High levels of serum transaminases, lymphocyte infiltration, some Kupffer's cell prominence, and extensive single cell hepatocyte death were found in AdCMVtk/GCV-treated animals, indicating severe liver damage induced, as expected, by the noncell-specific CMV promoter. XTL-1 cells (2 x 10(6)) were injected sc into BALB/c-severe combined immunodeficient mice (BALB/c-SCID), and the mice developed tumors after 3 wk. After intratumoral injection of AdrTgtk and treatment with GCV, tumors stabilized in 15 of 17 mice within 3 wk, 9 tumors remained stabilized after 5 wk of treatment, and 2 disappeared during observation. In AdCMVLuc/GCV-treated control mice, almost all tumors grew continuously. The average tumor size in AdrTgtk treated mice was significantly smaller than that of control animals after 2 wk of treatment. Our data confirm the effectiveness and specificity of an adenovirus using rTg promoter to express TK, and support its future application to thyroid cancer gene therapy in humans. PMID- 11889219 TI - Insulin inhibits the pro-inflammatory transcription factor early growth response gene-1 (Egr)-1 expression in mononuclear cells (MNC) and reduces plasma tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) concentrations. AB - We have recently demonstrated that an infusion of a low dose of insulin reduces the intranuclear NF-kappa B (a pro-inflammatory transcription factor) content in MNC while also reducing the plasma concentration of NF-kappa B dependent pro inflammatory cytokines and adhesion molecules. We have now tested the effect of insulin on the pro-inflammatory transcription factor, early growth response-1 (Egr-1) and plasma concentration of tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1), two major proteins whose expression is modulated by Egr-1. Insulin was infused at the rate of 2 IU/h in 5% dextrose (100 mL/h) and KCI (8 mmol/h) for 4 h in the fasting state in ten obese subjects. Blood samples were obtained at 0, 2, 4 and 6 h. MNC were isolated and their total homogenates and nuclear fractions were prepared and Egr-1 was measured by electrophoretic mobility shift assay (EMSA). Plasma TF and PAI-1 were assayed by ELISA. There was a significant fall in Egr-1 at 2 (66 +/- 14% of basal level) and 4 h (47 +/- 17% of the basal level; P<0.01). PAI-1 levels (basal = 100%) decreased significantly after insulin infusion at 2 h (57 +/- 6.7% of the basal level) and at 4 h (58 +/- 8.3% of the basal level; P<0.001). Plasma TF levels (basal = 100%) decreased to 76 +/- 7.7% of the basal level at 2 h and to 85 +/- 10.4% of the basal level at 4 h (P<0.05). Thus, insulin reduces intranuclear Egr-1 and the expression of TF and PAI-1. These data provide further evidence that insulin has an anti-inflammatory effect including the inhibition of TF and PAI-1 expression. These effects suggest a potential beneficial effect of insulin in thrombin formation and fibrinolysis in atherothrombosis. PMID- 11889218 TI - Presence of functional oxytocin receptors in cultured human myoblasts. AB - In the present report, we provide for the first time evidence that functional oxytocin receptors (OTRs) are present in human myoblasts obtained from clonal cultures of postnatal satellite cells. First, binding studies performed with a non selective vasopressin (AVP) and oxytocin (OT) radioligand indicated the presence of a single class of binding sites. Second, OTR mRNA was detected by RT PCR analysis whereas transcripts for AVP V(1a), V(1b) or V(2) receptors (V(1a)R, V(1b)R and V(2)R respectively) were not detected. Third, the presence of functional OTRs was evidenced by showing that agonist substances having a high affinity for the human OTR, namely OT, AVP and [Thr(4)Gly(7)]OT, increased the rate of myoblasts fusion and myotubes formation in the cultures, whereas F180, a V(1a)R selective agonist, and dDAVP, a V(2)R agonist had no significant effect on the fusion process. In addition, we show by RT-PCR and immunocytochemistry that the OT gene is expressed in cultured myoblasts. Taken together, our data suggest that OT may act as a paracrine/autocrine agent that stimulates the fusion of human myoblasts in vitro. In vivo, OT may be involved in the differentiation of human skeletal muscle during postnatal growth, and possibly its regeneration following injury. PMID- 11889220 TI - A novel melanocortin 3 receptor gene (MC3R) mutation associated with severe obesity. AB - The melanocortin 3 receptor (MC3R) plays a critical role in weight regulation as demonstrated in mouse models. We describe a novel mutation Ile183Asn (T548A) found in heterozygosity in a 13-year-old obese girl and her father. METHODS: The MC3R gene was sequenced in 41 unrelated obese children, and 121 DNA samples from non-obese individuals were analysed for this novel sequence variant by allele specific polymerase chain reaction (PCR). The genotypes of four family members of the pedigree were also analysed by allele-specific PCR. RESULTS: Ile183Asn was found in the proband and her father, though all four family members were obese. The sequence variant was not found in 121 control samples. The proband has high percentage body fat (49%), but the father's percentage body fat was only 30%. There were no distinguishing phenotypic features. Insulin sensitivity was significantly higher compared to the 40 other obese subjects without MC3R gene mutations. DISCUSSION: The difference in phenotypes between the two related heterozygotes, and the observation of obesity in other family members without the mutation suggests that obesity results from a varying combination of environmental, behavioural and multiple genetic factors (other than MC3R), even within the same family. PMID- 11889221 TI - Expression of gonadotropin-releasing hormone II (GnRH-II) receptor in human endometrial and ovarian cancer cells and effects of GnRH-II on tumor cell proliferation. AB - Recently it was shown that a second GnRH system exists in primates. This study was conducted to investigate whether or not the receptor specific for GnRH type II is expressed in human endometrial and ovarian cancer cells and whether or not GnRH-II has effects on tumor cell proliferation. Expression of GnRH-II receptor mRNA in endometrial and ovarian cancer cell lines was demonstrated using RT-PCR and Southern blot analysis. The proliferation of these cell lines was dose- and time-dependently reduced by native GnRH-II. These effects were significantly more potent than the anitproliferative effects of equimolar doses of GnRH-I agonist Triptorelin (p<0.001). In the GnRH-II receptor positive but GnRH-I receptor negative ovarian cancer cell line SK-OV-3 native GnRH-II but not GnRH-I agonist Triptorelin had antiproliferative effects. PMID- 11889222 TI - Ovarian function in an adult woman with McCune-Albright syndrome. PMID- 11889224 TI - Endocrine-related resources from the National Institutes of Health. PMID- 11889225 TI - National Hormone & Peptide Program--NIDDK: Recombinant hormones, hypothalamic peptides & other hormones, antisera, reagents, & hormone assay services available. PMID- 11889226 TI - We need something better, and we need it now: fetal striatal transplantation in Huntington's disease? PMID- 11889227 TI - B cells and epilepsy: the odd couple. PMID- 11889228 TI - Clinical trial end points: on the road to nowhere? AB - Over the past decade, there have been an increasing number of therapeutic clinical trials of PD. Despite many of these trials showing a positive treatment effect, few have resulted in clear and unambiguous recommendations regarding clinical practice, unlike trials of cerebrovascular disease. The authors hypothesize that lingering therapeutic uncertainty exists because many of the clinical trial end points have been surrogate outcome measures rather than end points with clear and convincing value to patients. The theoretical advantage of using validated surrogate outcomes in definitive trials includes smaller, faster, and less expensive studies. Consequences of using surrogate outcomes that have not been validated include ambiguous evidence and wasted resources as well as patient harm and missed opportunities. To optimize the chance that future trial results will provide clear treatment verdicts and to take full advantage of exciting developments in biomarker technology such as SPECT and PET, it is becoming progressively more urgent to understand the proper role, use, and challenges of using surrogate outcome measures in PD therapeutics. PMID- 11889229 TI - Bilateral human fetal striatal transplantation in Huntington's disease. AB - BACKGROUND: Transplanted striatal cells have been demonstrated to survive, grow, establish afferent and efferent connections, and improve behavioral signs in animal models of Huntington's disease (HD). OBJECTIVE: To evaluate feasibility and safety and to provide preliminary information regarding the efficacy of bilateral human fetal striatal transplantation in HD. METHODS: Seven symptomatic patients with genetically confirmed HD underwent bilateral stereotactic transplantation of two to eight fetal striata per side in two staged procedures. Tissue was dissected from the lateral half of the lateral ventricular eminence of donors 8 to 9 weeks postconception. Subjects received cyclosporine for 6 months. RESULTS: Three subjects developed subdural hemorrhages (SDHs) and two required surgical drainage. One subject died 18 months after surgery from probable cardiac arrhythmia secondary to severe atherosclerotic cardiac disease. Autopsy demonstrated clearly demarcated grafts of typical developing striatal morphology, with host-derived dopaminergic fibers extending into the grafts and no evidence of immune rejection. Other adverse events were generally mild and transient. Mean Unified HD Rating Scale (UHDRS) motor scores were 32.9 plus minus 6.2 at baseline and 29.7 plus minus 7.5 12 months after surgery (p = 0.24). Post-hoc analysis, excluding one subject who experienced cognitive and motor deterioration after the development of symptomatic bilateral SDHs, found that UHDRS motor scores were 33.8 plus minus 6.2 at baseline and 27.5 plus minus 5.2 at 12 months (p = 0.03). CONCLUSIONS: Transplantation of human fetal striatal cells is feasible and survival of transplanted cells was demonstrated. Patients with moderately advanced HD are at risk for SDH after transplantation surgery. PMID- 11889230 TI - Regional and progressive thinning of the cortical ribbon in Huntington's disease. AB - BACKGROUND: Huntington's disease (HD) is a fatal and progressive neurodegenerative disease that is accompanied by involuntary movements, cognitive dysfunction, and psychiatric symptoms. Although progressive striatal degeneration is known to occur, little is known about how the disease affects the cortex, including which cortical regions are affected, how degeneration proceeds, and the relationship of the cortical degeneration to clinical symptoms. The cortex has been difficult to study in neurodegenerative diseases primarily because of its complex folding patterns and regional variability; however, an understanding of how the cortex is affected by the disease may provide important new insights into it. METHODS: Novel automated surface reconstruction and high-resolution MR images of 11 patients with HD and 13 age-matched subjects were used to obtain cortical thickness measurements. The same analyses were performed on two postmortem brains to validate these methods. RESULTS: Regionally specific heterogeneous thinning of the cortical ribbon was found in subjects with HD. Thinning occurred early, differed among patients in different clinical stages of disease, and appeared to proceed from posterior to anterior cortical regions with disease progression. The sensorimotor region was statistically most affected. Measurements performed on MR images of autopsy brains analyzed similarly were within 0.25 mm of those obtained using traditional neuropathologic methods and were statistically indistinguishable. CONCLUSIONS: The authors propose that the cortex degenerates early in disease and that regionally selective cortical degeneration may explain the heterogeneity of clinical expression in HD. These measures might provide a sensitive prospective surrogate marker for clinical trials of neuroprotective medications. PMID- 11889231 TI - Spinocerebellar ataxias in the Netherlands: prevalence and age at onset variance analysis. AB - BACKGROUND: International prevalence estimates of autosomal dominant cerebellar ataxias (ADCA) vary from 0.3 to 2.0 per 100,000. The prevalence of ADCA in the Netherlands is unknown. Fifteen genetic loci have been identified (SCA-1-8, SCA 10-14, SCA-16, and SCA-17) and nine of the corresponding genes have been cloned. In SCA-1, SCA2, SCA3, SCA6, SCA7, SCA-12 and SCA-17 the mutation has been shown to be an expanded CAG repeat. Previously, the length of the CAG repeat was found to account for 50 to 80% of variance in age at onset. Because of heterogeneity in encoded proteins, different pathophysiologic mechanisms leading to neurodegeneration could be involved. The relationship between CAG repeat length and age at onset would then differ accordingly. METHOD: Based on the results of SCA mutation analysis in the three DNA diagnostic laboratories that serve the entire Dutch population, the authors surveyed the number of families and affected individuals per SCA gene, as well as individual repeat length and age at onset. Regression analysis was applied to study the relationship between CAG repeat length and age at onset per SCA gene. The slopes of the different regression curves were compared. RESULTS: On November 1, 2000, mutations were found in 145 ADCA families and 391 affected individuals were identified. The authors extrapolated a minimal prevalence of 3.0 per 100,000 (range 2.8 to 3.8/100,000). SCA3 was the most frequent mutation. CAG repeat length contributed to 52 to 76% of age at onset variance. Regression curve slopes for SCA-1, SCA2, SCA3, and SCA7 did not differ significantly. CONCLUSIONS: The estimated minimal prevalence of ADCA in the Netherlands is 3.0 per 100,000 inhabitants. Except for SCA6, the relationship between age at onset and CAG repeat expansion does not differ significantly between SCA-1, SCA2, SCA3, and SCA7 patient groups in our population, indicating that these SCA subtypes share similar mechanisms of polyglutamine-induced neurotoxicity, despite heterogeneity in gene products. PMID- 11889232 TI - Analysis of antibody gene rearrangement, usage, and specificity in chronic focal encephalitis. AB - BACKGROUND: Autoantibodies have been implicated in the development of chronic focal encephalitis (CFE) or Rasmussen's disease, a progressive and intractable form of epilepsy characterized by uncontrollable unilateral focal seizures, brain atrophy, and inflammation. OBJECTIVE: To investigate the origin and characteristics of the B cell population that trigger or sustain brain inflammation in patients with CFE. METHODS: The authors used immunoglobulin (Ig) complementary determining region 3 (CDR3)-size spectratyping and DNA sequencing to examine the rearranged IgG heavy chain (IgGH) transcript repertoire in resected brain samples from four patients with CFE. They also performed Western blotting on human and rat brain homogenates and immunostaining on a human neuronal cell line to test the reactivity of sera from patients with CFE. RESULTS: The authors observed substantial perturbations from the normal, unstimulated repertoire of immunoglobulin genes. Sequencing of randomly selected clones confirmed the restricted profile and provided evidence for somatic mutation patterns characteristic of antigen-specific stimulation. They also observed IgGVH-CDR3 sequence diversity among patients. When sera were assayed from patients with CFE for specificity against rat and human brain homogenates, heterogeneous reactivity patterns were detected among patients. Immunostaining of postmitotic human neuronal cells demonstrated reactivity of some patients' sera against neural antigens. CONCLUSIONS: These findings support an important role for clonally expanded B lymphocytes in some forms of epilepsy, but also indicate a wide spectrum of reactivity characteristic of antigenic heterogeneity. PMID- 11889233 TI - Low incidence of abnormal (18)FDG-PET in children with new-onset partial epilepsy: a prospective study. AB - OBJECTIVE: Patients with refractory partial epilepsy often exhibit regional hypometabolism. It is unknown whether the metabolic abnormalities are present at seizure onset or develop over time. METHODS: The authors studied 40 children within 1 year of their third unprovoked partial seizure with EEG, MRI, and [(18)F]-fluorodeoxyglucose ((18)FDG)-PET (mean age at seizure onset = 5.8 years, range 0.9 to 11.9 years; mean epilepsy duration = 1.1 years, range 0.3 to 2.3 years; mean number of seizures = 30, range 3 to 200). The authors excluded children with abnormal structural MRI, except four with mesial temporal sclerosis and two with subtle hippocampal dysgenesis. (18)FDG-PET was analyzed with a region of interest template. An absolute asymmetry index, [AI], greater than 0.15 was considered abnormal. RESULTS: Thirty-three children had a presumptive temporal lobe focus, five frontotemporal, and two frontal. Mean AI for all regions was not different from 10 normal young adults, even when children less likely to have a temporal focus were excluded. Eight of 40 children (20%) had focal hypometabolism, all restricted to the temporal lobe, especially inferior mesial and inferior lateral regions. Abnormalities were ipsilateral to the presumed temporal lobe ictal focus. CONCLUSIONS: Abnormalities of glucose utilization may be less common and profound in children with new-onset partial seizures than in adults with chronic partial epilepsy. Although these patients' prognosis is uncertain, resolution of epilepsy after three documented seizures is uncommon. If the subjects develop a higher incidence of hypometabolism in the future with planned follow-up studies, metabolic dysfunction may be related to persistent epilepsy rather than present at seizure onset. PMID- 11889234 TI - Clinical, EEG, and quantitative MRI differences in pediatric frontal and temporal lobe epilepsy. AB - OBJECTIVE: To examine the clinical, electrographic, and quantitative MRI differences between frontal lobe (FLE) and mesial temporal lobe epilepsy (MTLE) in children. METHODS: The population included children who underwent video-EEG monitoring between 1995 and 2000 who were classified as either FLE (n = 39) or MTLE (n = 17) according to the criteria of the International League Against Epilepsy. Clinical, EEG, and quantitative MRI data (including frontal cortical volumes) were compared between the two syndromes and a control group (n = 42). RESULTS: In FLE, seizures were significantly briefer, more frequent, and predominantly from sleep, and had differing motor characteristics. The rates of bilateral epileptiform interictal and ictal EEG abnormalities were significantly higher in FLE. A nonlesional MRI was significantly more common in FLE. Mean frontal cortical volume in FLE was significantly lower than MTLE and controls. Seizure freedom after surgery was lower in FLE. CONCLUSIONS: The clinical syndrome of FLE is clearly distinct from MTLE. The etiology of this disorder is unknown in the majority of cases despite extensive investigation. Because of a lack of a clearly defined etiology and frequent nonlateralizing EEG changes, few of these children are considered optimal surgical candidates. The demonstration of bilateral frontal cortical volume loss and bilateral EEG abnormalities suggests that FLE is a bilateral disease in a high proportion of patients. The outcome in those patients who were deemed surgical candidates was significantly worse than the MTLE cases. PMID- 11889235 TI - Abnormal cobalamin-dependent transmethylation in AIDS-associated myelopathy. AB - BACKGROUND: White matter vacuolization of the spinal cord is common in patients with AIDS and may lead to clinical manifestations of myelopathy. The pathogenesis of AIDS-associated myelopathy (AM) is unknown and may be related to metabolic abnormalities rather than to direct HIV infection. The striking pathologic similarity between AM and the vacuolar myelopathy associated with vitamin B(12) deficiency suggests that abnormal metabolism of the B(12)-dependent transmethylation pathway may be important in the pathogenesis of AM. METHODS: The authors compared S-adenosyl-methionine (SAM), methionine, homocysteine, and glutathione in serum and CSF of 15 patients with AM vs. 13 HIV-infected controls without myelopathy (HWM). They also compared the results with a non-HIV--infected reference population (NC). All patients had normal B(12), folate, and methylmalonic acid levels. RESULTS: There was a decrease in CSF SAM in the AM group compared with the HWM group (p < 0.0001) and the NC group (p < 0.0001). CSF SAM in the HWM group was also lower than that in the NC group (p = 0.015). Serum methionine was also reduced in serum of the myelopathic group compared with the NC group (p = 0.006). CONCLUSIONS: AM is associated with an abnormality of the vitamin B(12)-dependent transmethylation pathway. PMID- 11889237 TI - Does dichotic listening probe temporal lobe functions? AB - OBJECTIVE: To explore cortical hemodynamic responses using fMRI in the context of three dichotic listening tasks. BACKGROUND: Dichotic listening is a widely used behavioral technique indicating brain laterality during which subjects are presented with two different auditory signals at the same time, one arriving at each ear. fMRI offers the potential to explore the hemodynamic response during dichotic listening and to relate the behavioral indices with these cortical measures. METHOD: fMRI was performed for 10 right-handed normal subjects listening to consonant-vowel syllable pairs with the task of detecting a "target" syllable by pressing a button. The target stimulus appeared equally often in the left and right ear. The subjects were instructed to either concentrate on the stimuli presented in both ears (DIV) or only in the left ear (FL) or right ear (FR). In addition, a control condition was used during which the syllables were presented binaurally. Hemodynamic responses were measured by applying whole-head echo planar imaging techniques and statistically analyzed by using statistical parametric mapping (SPM99) software. RESULTS: During dichotic listening, there were generally extended activations in frontotemporal networks. For the DIV condition, the authors found strong bilateral activations in the inferior frontal gyrus, Broca's area, the left middle frontal gyrus, and in the left superior temporal gyrus. During the FL condition, there was an additional cluster in the right inferior frontal gyrus. For the FR condition, there were stronger activations in Broca's area and the left superior temporal gyrus. CONCLUSIONS: These findings were taken as evidence that dichotic listening is more demanding, requiring more processing capacity distributed in frontotemporal networks. The behavioral measures of dichotic listening were not simply a function of temporal lobe activation. Rather, the cortical activations support the notion that different processing strategies controlled by different neural structures are applied during dichotic listening. PMID- 11889238 TI - The effect of APOE epsilon4 allele on cerebral glucose metabolism in AD is a function of age at onset. AB - BACKGROUND: Although the APOE epsilon4 allele is a well-known risk factor for developing AD, the impact of the epsilon4 allele on clinical manifestations in patients with AD is still controversial. One possible reason for this controversy is that previous studies did not consider the effect of patient age at symptom onset. OBJECTIVE: To investigate the possible impact of patient age at onset of AD on the effect of APOE genotype on regional cerebral glucose metabolism (rCMRglc). METHODS: The authors compared rCMRglc between probable AD patients (based on criteria of the National Institute of Neurologic Disease and Stroke/AD and Related Disorders Association) with APOE epsilon4/4 and APOE epsilon3/3 alleles in early-onset (< or =65 years old) and late-onset (>65 years old) groups. In each group, the patients with APOE epsilon4/4 and APOE epsilon3/3 alleles were comparable for age at onset, age at examination, sex, disease duration, education level, and severity of dementia. RESULTS: In the early-onset group, the patients with the APOE epsilon4/4 genotype showed a significant decrease of rCMRglc in the medial temporal lobe and a significant increase of rCMRglc in the inferior parietal and posterior temporal cortices as compared with those patients with the APOE epsilon3/3 genotype. In the late-onset group, there were no significant differences in the rCMRglc pattern between the patients with APOE epsilon4/4 and APOE epsilon3/3 alleles. CONCLUSIONS: The current findings indicate that the impact of the APOE epsilon4 genotype on cerebral glucose metabolism of patients with AD may be a function of age at symptom onset. PMID- 11889239 TI - Antemortem MRI findings correlate with hippocampal neuropathology in typical aging and dementia. AB - OBJECTIVES: To assess the diagnostic specificity of MRI-defined hippocampal atrophy for AD among individuals with a variety of pathologically confirmed conditions associated with dementia as well as changes attributable to typical aging, and to measure correlations among premortem MRI measurements of hippocampal atrophy, mental status examination performance, and the pathologic stage of AD. METHODS: An unselected series of 67 individuals participating in the Mayo Alzheimer's Disease Research Center/Alzheimer's Disease Patient Registry who had undergone a standardized antemortem MRI study and also postmortem examination were identified. Hippocampal volumes were measured from antemortem MRI. Each postmortem specimen was assigned a pathologic diagnosis and in addition, the severity of AD pathology was staged using the method of Braak and Braak. RESULTS: Individuals with an isolated pathologic diagnosis of AD, hippocampal sclerosis, frontotemporal degeneration, and neurofibrillary tangle--only degeneration usually had substantial hippocampal atrophy, while those with changes of typical aging did not. Among all 67 subjects, correlations (all p < 0.001) were observed between hippocampal volume and Braak and Braak stage (r = -0.39), between hippocampal volume and Mini-Mental State Examination (MMSE) score (r = 0.60), and between MMSE score and Braak and Braak stage (r = -0.41). CONCLUSIONS: Hippocampal atrophy, while not specific for AD, was a fairly sensitive marker of the pathologic AD stage (particularly among subjects with isolated AD pathology [r = -0.63, p = 0.001]) and consequent cognitive status. PMID- 11889240 TI - Functional deficits in patients with mild cognitive impairment: prediction of AD. AB - OBJECTIVE: To evaluate the predictive utility of self-reported and informant reported functional deficits in patients with mild cognitive impairment (MCI) for the follow-up diagnosis of probable AD. METHODS: The Pfeffer Functional Activities Questionnaire (FAQ) and Lawton Instrumental Activities of Daily Living (IADL) Scale were administered at baseline. Patients were followed at 6-month intervals, and matched normal control subjects (NC) were followed annually. RESULTS: Self-reported deficits were higher for patients with MCI than for NC. At baseline, self- and informant-reported functional deficits were significantly greater for patients who converted to AD on follow-up evaluation than for patients who did not convert, even after controlling for age, education, and modified Mini-Mental State Examination scores. While converters showed significantly more informant- than self-reported deficits at baseline, nonconverters showed the reverse pattern. Survival analyses further revealed that informant-reported deficits (but not self-reported deficits) and a discrepancy score indicating greater informant- than self-reported functional deficits significantly predicted the development of AD. The discrepancy index showed high specificity and sensitivity for progression to AD within 2 years. CONCLUSIONS: These findings indicate that in patients with MCI, the patient's lack of awareness of functional deficits identified by informants strongly predicts a future diagnosis of AD. If replicated, these findings suggest that clinicians evaluating MCI patients should obtain both self-reports and informant reports of functional deficits to help in prediction of long-term outcome. PMID- 11889241 TI - ALS and PDC of Guam: forty-year follow-up. AB - BACKGROUND: It was noticed in the mid-1950s that the incidence of ALS and parkinsonism--dementia complex (PDC) were much higher on Guam than anywhere else in the world. In 1958, a registry of patients and controls was established to ascertain the familial and genetic aspects of these diseases. Patients and individually matched controls and their relatives were registered from 1958 to 1963. The registry was updated and analyzed in 1998 through 1999. OBJECTIVE: To ascertain whether first-degree relatives of patients had a higher risk for developing ALS or PDC than relatives of controls. METHODS: During the period of 1958 to 1963, 126 new patients and 126 individually matched controls and their respective first-degree relatives and spouses were evaluated neurologically and registered. Forty years later, the number of new cases among the patient and control relatives were compared to an expected number of new cases based on the age- and sex-specific incidence of ALS and PDC in the population at large. RESULTS: From 1958 to 1999, there were 102 new ALS or PDC cases among relatives of patients and 33 among relatives of controls. These values were compared with the derived expected values. There were more observed than expected new cases among patients' relatives, and less observed cases than expected among the controls' relatives. CONCLUSIONS: Relatives of patients with ALS or PDC have significantly higher risks for developing the disease than the Guamanian population, whereas relatives of controls have significantly lower risks. PMID- 11889244 TI - Effect of time since onset of risk factors on the occurrence of ischemic stroke. AB - OBJECTIVE: To determine the effect of time since onset of risk factors on the modeling of risk factors for ischemic stroke. METHODS: The resources of the Rochester Epidemiology Project allowed identification of the 1,397 incident cases of ischemic stroke and age- and sex-matched control subjects from the population for 1970 through 1989. These cases and controls permitted the development of a multiple conditional logistic regression model to estimate the odds ratios of ischemic stroke for various risk factors. The time since onset variables for each risk factor were then added to the model to determine which were significant and to assess their impact on variables in the model. RESULTS: The time since onset variables for congestive heart failure and TIA were the only variables of this type included in the resultant model. Each showed the highest risk for stroke soon after the onset of the risk factor. In addition, the influence of congestive heart failure was higher at younger ages. Hypertension (with or without left ventricular hypertrophy) increases the risk for stroke but has a diminishing influence with increasing age. In addition, persons with left ventricular hypertrophy are at a higher risk than those with hypertension alone, although this difference also decreases with age. The time since onset variables pertaining to systolic hypertension at 140 to 159 mm Hg, 160 to 179 mm Hg, and > or =180 mm Hg were not significant in any analysis. CONCLUSIONS: TIA and congestive heart failure were the only risk factors for stroke for which time since onset was significant in the model for predicting ischemic stroke. PMID- 11889242 TI - Early detection and longitudinal changes in amyotrophic lateral sclerosis by (1)H MRSI. AB - OBJECTIVE: To determine 1) the reproducibility of metabolite measurements by (1)H MRS in the motor cortex; 2) the extent to which (1)H MRS imaging (MRSI) detects abnormal concentrations of N-acetylaspartate (NAA)-, choline (Cho)-, and creatine (Cre)-containing compounds in early stages of ALS; and 3) the metabolite changes over time in ALS. METHODS: Sixteen patients with definite or probable ALS, 12 with possible or suspected ALS, and 12 healthy controls underwent structural MRI and multislice (1)H MRSI. (1)H MRSI data were coregistered with tissue-segmented MRI data to obtain concentrations of NAA, Cre, and Cho in the left and right motor cortex and in gray matter and white matter of nonmotor regions in the brain. RESULTS: The interclass correlation coefficient of NAA was 0.53 in the motor cortex tissue and 0.83 in nonmotor cortex tissue. When cross-sectional data for patients were compared with those for controls, the NAA/(Cre + Cho) ratio in the motor cortex region was significantly reduced, primarily due to increases in Cre and Cho and a decrease in NAA concentrations. A similar, although not significant, trend of increased Cho and Cre and reduced NAA levels was also observed for patients with possible or suspected ALS. Furthermore, in longitudinal studies, decreases in NAA, Cre, and Cho concentrations were detected in motor cortex but not in nonmotor regions in ALS. CONCLUSION: Metabolite changes measured by (1)H MRSI may provide a surrogate marker of ALS that can aid detection of early disease and monitor progression and treatment response. PMID- 11889243 TI - Myosin heavy chain IIa gene mutation E706K is pathogenic and its expression increases with age. AB - BACKGROUND: The authors recently described a new autosomal dominant myopathy (OMIM 605637 inclusion body myopathy 3) associated with a missense mutation in the myosin heavy chain (MyHC) IIa gene (MyHC IIa, Human Gene Map [HGM] locus MYH2). Young patients showed minor changes in their muscle biopsies, although dystrophic alterations and rimmed vacuoles with 15- to 20-nm tubulofilaments identical to those in sporadic inclusion body myositis (s-IBM) were observed in some of the adult (especially older) patients. The current study was undertaken to investigate the relation between expression of the mutant MyHC IIa and pathologic changes in muscle. METHODS: The expression of MyHC IIa in nine muscle specimens from six individuals carrying the mutation was analyzed by immunohistochemistry, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and a new reverse transcriptase--PCR method to measure the relative abundance of the various MyHC transcripts. RESULTS: Young patients with muscle weakness and minor pathologic changes in muscle expressed MyHC IIa at undetectable levels. MyHC IIa was expressed at high levels in adults with a progressive clinical course and dystrophic muscle changes. In these cases, a large number of muscle fibers were hybrids with expression of more than one MyHC isoform. Both MyHC IIa alleles were equally expressed. The relative level of MyHC IIa transcripts exceeded that of the corresponding protein, indicating an increased turnover of mutated protein. MyHC IIa expression was a consistent finding in muscle fibers with rimmed vacuoles. CONCLUSIONS: The clear correlation between pathologic changes and expression of MyHC IIa indicates that defects in MyHC may lead not only to muscle weakness but also to muscle degeneration. The consistent expression of MyHC IIa in muscle fibers with rimmed vacuoles indicates that the breakdown of sarcomeric proteins is a key element in the pathogenesis of rimmed vacuoles of s-IBM type. PMID- 11889245 TI - Adaptation in the motor cortex following cervical spinal cord injury. AB - BACKGROUND: The nature of the adaptive changes that occur in the cerebral cortex following injury to the cervical spinal cord are largely unknown. OBJECTIVE: To investigate these adaptive changes by examining the relationship between the motor cortical representation of the paretic right upper extremity compared with that of the tongue. The tongue was selected because the spinal cord injury (SCI) does not affect its movement and the cortical representation of the tongue is adjacent to that of the paretic upper extremity. METHODS: FMRI was used to map cortical representations associated with simple motor tasks of the right upper extremity and tongue in 14 control subjects and 9 patients with remote (>5.5 months) cervical SCI. RESULTS: The mean value for the site of maximum cortical activation during upper limb movement was identical between the two groups. The site of maximum left hemispheric cortical activation during tongue movement was 12.8 mm (p < 0.01) medial and superior to that of control subjects, indicating the presence of a shift in cortical activation. CONCLUSION: The findings indicate that the adult motor cortex does indeed adapt following cervical SCI. The nature of the adaptation and the underlying biological mechanisms responsible for this change require further investigation. PMID- 11889246 TI - Trochleitis and migraine headache. AB - Idiopathic trochleitis is a cause of superimposed ocular pain in patients with migraine. Trochleitis usually presents as an orbital pain without obvious ocular signs. Like greater occipital neuralgia, trochleitis may sustain or trigger the pain of chronic migraine. Diagnosis is confirmed by peritrochlear steroid injection, which produces a quick relief of periocular symptoms and may improve headache control. PMID- 11889247 TI - Accuracy of muscle localization without EMG: implications for treatment of limb dystonia. AB - Although botulinum toxin is an effective treatment for focal dystonia, the importance of electromyography (EMG) in identifying muscles and guiding injections is unclear. The authors examined the accuracy of muscle localization in 38 muscles in patients with focal hand dystonia without EMG guidance. Only 37% of needle placement attempts reached the target muscles or muscle fascicles. This study demonstrates that EMG guidance is needed for correct localization of desired muscles. PMID- 11889248 TI - Parkin mutations in a patient with hemiparkinsonism-hemiatrophy: a clinical genetic and PET study. AB - The authors describe a 37-year-old woman with early-onset hemiparkinsonism (HP) and ipsilateral body hemiatrophy (HA). Genetic analysis revealed a missense mutation (Arg275Trp) and a duplication of exon 7 of parkin. The complementary metabolic and receptor pattern of PET ligands corresponded to that typically found in idiopathic PD, although tracer binding asymmetry was lacking. Parkin mutations should be considered in HPHA, particularly when there is a younger age at onset and dystonia is an early sign. PMID- 11889250 TI - Spontaneous CSF leaks: underlying disorder of connective tissue. AB - Of 58 consecutive patients with spontaneous CSF leaks, nine exhibited features of connective tissue disorder. One had Marfan's syndrome. Five additional patients had hyperflexible joints, of whom four had arachnodactyly, four were tall and slender, two had hyperextensible skin, and one had a strong family history of abdominal aorta aneurysms. Retinal detachment at a young age was noted in two. One patient had bilateral carotid dissections. A dural weakness may predispose patients to spontaneous CSF leak. PMID- 11889249 TI - Early-onset, rapidly progressive familial tauopathy with R406W mutation. AB - An early-onset and rapidly progressive familial tauopathy with R406W mutation is described. The patient was a 47-year-old man who first presented with psychiatric symptoms followed by overt dementia at age 52 and died 1 year later. Postmortem study revealed tangle-associated neuronal degeneration, accentuated in the medial temporal lobe. R406W mutation was determined by sequence analysis and immunocytochemically with anti-mutant tau antibody. PMID- 11889251 TI - Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy. AB - The clinicopathologic features of two Japanese sisters with cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) are described. Neither patient had a history of hypertension, and both experienced cerebrovascular events before reaching their forties. Severe degenerative changes in the lumbar spine and knee joints were seen on radiographs. MRI showed extensive cerebral white matter lesions, which revealed remarkable arteriosclerotic changes on autopsy. PMID- 11889252 TI - 1H MRSI predicts surgical outcome in MRI-negative temporal lobe epilepsy. AB - 1H MRS imaging (MRSI) was performed on 15 patients with MRI-negative temporal lobe epilepsy (TLE) who underwent seizure surgery. The non-seizure-free patients (NSF) ipsilateral hippocampal N-acetylaspartate (NAA)/(Cr+Cho) z scores were lower than the contralateral scores (p = 0.04), and the NSF ipsilateral z scores were lower than the seizure-free patients' (SF) ipsilateral z scores (p = 0.0049). Similarly, NSF contralateral scores were lower than contralateral SF (p = 0.02). These findings suggest NAA predicts the surgical outcome in patients with TLE without evidence of mesial temporal sclerosis on MRI. PMID- 11889253 TI - Intrathecal IgM synthesis in neurologic diseases: relationship with disability in MS. AB - The authors studied the intrathecal IgM synthesis (ITMS) in paired sera and CSF samples from 65 patients with MS, 28 with CNS infection, 40 with other neurologic diseases and eight control subjects. ITMS was found in 30 patients with MS and in 20 with CNS infection, but not in patients with other neurologic diseases or in control subjects. In infectious samples, the ITMS is likely a primary response. In MS group, it was associated with higher Expanded Disability Status Scale index (p = 0.017). PMID- 11889254 TI - Effects of L-arginine on the acute phase of strokes in three patients with MELAS. PMID- 11889255 TI - Association of the 5-HT6 receptor gene polymorphism C267T with Parkinson's disease. PMID- 11889256 TI - Alopecia induced by dopamine agonists. PMID- 11889257 TI - Lack of effect of topiramate for central pain. PMID- 11889258 TI - Cabergoline can increase penile erections and libido. PMID- 11889260 TI - Fiction: the last referral. PMID- 11889261 TI - A 1-year, randomized, placebo-controlled study of donepezil in patients with mild to moderate AD. PMID- 11889262 TI - The parkin gene is not involved in late-onset Parkinson's disease. PMID- 11889263 TI - Prognostic value of decremental responses to repetitive nerve stimulation in ALS patients. PMID- 11889264 TI - Association study of dopamine D2, D3 receptor gene polymorphisms with motor fluctuations in PD. PMID- 11889265 TI - Language and spatial attention can lateralize to the same hemisphere in healthy humans. PMID- 11889266 TI - Frequency of bowel movements and future risk of Parkinson's disease. PMID- 11889269 TI - Introduction. PMID- 11889267 TI - Differing patterns of temporal atrophy in Alzheimer's disease and semantic dementia. PMID- 11889270 TI - The meaning and value of prevention research. PMID- 11889271 TI - Extramural prevention research at the Centers for Disease Control and Prevention. PMID- 11889272 TI - Partnerships and coalitions for community-based research. PMID- 11889273 TI - The National Birth Defects Prevention Study. AB - The National Birth Defects Prevention Study was designed to identify infants with major birth defects and evaluate genetic and environmental factors associated with the occurrence of birth defects. The ongoing case-control study covers an annual birth population of 482,000 and includes cases identified from birth defect surveillance registries in eight states. Infants used as controls are randomly selected from birth certificates or birth hospital records. Mothers of case and control infants are interviewed and parents are asked to collect buccal cells from themselves and their infants for DNA testing. Information gathered from the interviews and the DNA specimens will be used to study independent genetic and environmental factors and gene-environment interactions for a broad range of birth defects. As of December 2000, 7,470 cases and 3,821 controls had been ascertained in the eight states. Interviews had been completed with 70% of the eligible case and control mothers, buccal cell collection had begun in all of the study sites, and researchers were developing analysis plans for the compiled data. This study is the largest and broadest collaborative effort ever conducted among the nation's leading birth defect researchers. The unprecedented statistical power that will result from this study will enable scientists to study the epidemiology of some rare birth defects for the first time. The compiled interview data and banked DNA of approximately 35 categories of birth defects will facilitate future research as new hypotheses and improved technologies emerge. PMID- 11889274 TI - The Tuberculosis Trials Consortium: a model for clinical trials collaborations. PMID- 11889275 TI - The contributions of managed care plans to public health practice: evidence from the nation's largest local health departments. AB - OBJECTIVE: The authors examine the extent and nature of managed care plans participating in local public health activities. METHODS: In 1998, the authors surveyed the directors of all US local health departments serving jurisdictions of at least 100,000 residents to collect information about public health activities performed in their jurisdictions and about organizations participating in the activities. Multivariate logistic and linear regression models were used to examine organizational and market characteristics associated with managed care plan participation in public health activities. RESULTS: Managed care plans were reported to participate in public health activities in 164 (46%) of the jurisdictions surveyed, and to contribute to 13% of the public health activities performed in the average jurisdiction. Plans appeared most likely to participate in public health activities involving the delivery or management of personal health services and the exchange of health-related information. Managed care participation was more likely to occur in jurisdictions with higher HMO penetration, fewer competing plans, and larger proportions of plans enrolling Medicaid recipients. Participation was positively associated with the overall scope and perceived effectiveness of local public health activities. CONCLUSIONS: Although plans participate in a narrow range of activities, these contributions may complement the work of public health agencies. PMID- 11889276 TI - Project Joy: faith based cardiovascular health promotion for African American women. AB - OBJECTIVE: The authors tested the impact on cardiovascular risk profiles of African American women ages 40 years and older after one year of participation in one of three church-based nutrition and physical activity strategies: a standard behavioral group intervention, the standard intervention supplemented with spiritual strategies, or self-help strategies. METHODS: Women were screened at baseline and after one year of participation. The authors analyzed intention-to treat within group and between groups using a generalized estimating equations adjustment for intra-church clustering. Because spiritual strategies were added to the standard intervention by participants themselves, the results from both active groups were similar and, thus, combined for comparisons with the self-help group. RESULTS: A total of 529 women from 16 churches enrolled. Intervention participants exhibited significant improvements in body weight (-1.1 lbs), waist circumference (-0.66 inches), systolic blood pressure (-1.6 mmHg), dietary energy (-117 kcal), dietary total fat (-8 g), and sodium intake (-145 mg). The self-help group did not. In the active intervention group, women in the top decile for weight loss at one year had even larger, clinically meaningful changes in risk outcomes (-19.8 lbs). CONCLUSIONS: Intervention participants achieved clinically important improvements in cardiovascular disease risk profiles one year after program initiation, which did not occur in the self-help group. Church-based interventions can significantly benefit the cardiovascular health of African American women. PMID- 11889277 TI - Safer choices: reducing teen pregnancy, HIV, and STDs. AB - OBJECTIVES: This study evaluated the long-term effectiveness of Safer Choices, a theory-based, multi-component educational program designed to reduce sexual risk behaviors and increase protective behaviors in preventing HIV, other STDs, and pregnancy among high school students. METHODS: The study used a randomized controlled trial involving 20 high schools in California and Texas. A cohort of 3869 ninth-grade students was tracked for 31 months from fall semester 1993 (baseline) to spring semester 1996 (31-month follow-up). Data were collected using self-report surveys administered by trained data collectors. Response rate at 31-month follow-up was 79%. RESULTS: Safer Choices had its greatest effect on measures involving condom use. The program reduced the frequency of intercourse without a condom during the three months prior to the survey, reduced the number of sexual partners with whom students had intercourse without a condom, and increased use of condoms and other protection against pregnancy at last intercourse. Safer Choices also improved 7 of 13 psychosocial variables, many related to condom use, but did not have a significant effect upon rates of sexual initiation. CONCLUSIONS: The Safer Choices program was effective in reducing important risk behaviors for HIV, other STDs, and pregnancy and in enhancing most psychosocial determinants of such behavior. PMID- 11889278 TI - Telehealth: reaching out to newly injured spinal cord patients. AB - OBJECTIVES: The authors present preliminary results on health-related outcomes of a randomized trial of telehealth interventions designed to reduce the incidence of secondary conditions among people with mobility impairment resulting from spinal cord injury (SCI). METHODS: Patients with spinal cord injuries were recruited during their initial stay at a rehabilitation facility in Atlanta. They received a video-based intervention for nine weeks, a telephone-based intervention for nine weeks, or standard follow-up care. Participants are followed for at least one year, to monitor days of hospitalization, depressive symptoms, and health-related quality of life. RESULTS: Health-related quality of life was measured using the Quality of Well-Being (QWB) scale. QWB scores (n = 111) did not differ significantly between the three intervention groups at the end of the intervention period. At year one post discharge, however, scores for those completing one year of enrollment (n = 47) were significantly higher for the intervention groups compared to standard care. Mean annual hospital days were 3.00 for the video group, 5.22 for the telephone group, and 7.95 for the standard care group. CONCLUSIONS: Preliminary evidence suggests that in-home telephone or video-based interventions do improve health-related outcomes for newly injured SCI patients. Telehealth interventions may be cost-saving if program costs are more than offset by a reduction in rehospitalization costs, but differential advantages of video-based interventions versus telephone alone warrant further examination. PMID- 11889279 TI - Long-term effectiveness of a peer-based intervention to promote condom and contraceptive use among HIV-positive and at-risk women. AB - OBJECTIVE: The authors used data from a larger study to evaluate the long-term effects of a peer advocate intervention on condom and contraceptive use among HIV infected women and women at high risk for HIV infection. METHODS: HIV-infected women in one study and women at high risk for HIV infection in a second study were selected from the Women and Infants Demonstration Project and assigned to a standard or an enhanced HIV prevention treatment group. The enhanced intervention included support groups and one-on-one contacts with peer advocates tailored to clients' needs. The authors interviewed women at baseline and at 6-, 12- and 18 months, and measured changes in consistency of condom and contraceptive use and in self-efficacy and perceived advantages and disadvantages of condom and contraceptive use. RESULTS: Of HIV-infected women, the enhanced group had improved consistency in condom use, increased perceived advantages of condom use, and increased level of self-efficacy compared with the standard group. Of women at risk, the enhanced intervention group at six months maintained consistent condom use with a main partner and perceived more benefit of condom use compared with the standard group. These differences diminished at 12 months. CONCLUSIONS: The enhanced intervention was generally effective in the HIV+ study. In the at risk study, however, intervention effects were minimal and short-lived. Factors related to the theory, intervention design, and sample characteristics help explain these differences. PMID- 11889280 TI - Maintenance of a smoking cessation program in public health clinics beyond the experimental evaluation period. AB - OBJECTIVES: As phase 3 of a study to evaluate a smoking cessation program in public health practice, the authors assess the maintenance and impact of the It's Time smoking cessation program in seven public maternal and child health clinics in Chicago. METHODS: The authors interviewed 404 clinic patients in the study's baseline phase (prior to introduction of the It's Time intervention program), and 610 in the program maintenance phase (in the year after experimental evaluation had ended) to assess exposure to smoking cessation interventions offered at the clinic, and smoking cessation outcomes (quit, actions toward quitting, scores on action, motivation, readiness, and confidence scales). The authors controlled for clustering of smokers within clinics, smokers' characteristics prior to clinic visit, and type of clinic service. They compared outcomes by study group (control or intervention) to which each clinic had been assigned in the earlier experimental phase. RESULTS: Compared to baseline, smokers in the maintenance phase had greater exposure to posters, provider advice and booklet, and better outcomes on seven of eight smoking cessation measures, including quitting. These improvements were larger for clinics with prior experience implementing It's Time. CONCLUSION: Participation in the experimental evaluation of the It's Time program prepared and possibly motivated the clinics to continue the program. Continuing the program resulted in greater delivery of interventions and improved smoking cessation outcomes for smokers in the clinics. PMID- 11889281 TI - Factors associated with adolescent initiation of injection drug use. AB - OBJECTIVE: The purpose of this study was to evaluate the extent to which demographic, sexual, and non-injection drug use practices predict adolescent initiation of injection drug use. METHODS: Street recruited injection drug users 15-30 years of age in Baltimore, Maryland, who initiated injection within five years of study enrollment, completed a questionnaire that included a year-by-year history regarding the five years prior to initiation of injection. Factors associated with initiation during adolescence (< or = 21 years of age) versus young adulthood (>21 ) were determined using logistic regression. RESULTS: Of 226 participants, most were female (61%) and African American (64%). Median age of participants was 25; median age at initiation of injection was 23. Factors significantly associated with adolescent initiation in multivariate analysis included race other than African American, and practices prior to initiating injection including condom use, lack of cocaine use, exclusive crack smoking just prior to initiation, and smoking marijuana. Adolescent initiates also had shorter durations of illicit drug use prior to initiating injection. CONCLUSION: Short term non-injection drug use, particularly exclusive crack smoking, was associated with adolescent initiation of injection drug use. Early prevention efforts targeting this high-risk group of younger drug users are warranted in order to delay or prevent onset of injection drug use. PMID- 11889282 TI - Risk factors for lyme disease in Chester County, Pennsylvania. AB - OBJECTIVE: To identify factors associated with increased or decreased risk of infection for Lyme disease in Chester County, Pennyslvania. METHODS: The authors designed an unmatched case-control study involving 294 incident cases reported to the Chester County Health Department in 1998 and 449 controls selected by random digit dialing. All case and control participants were interviewed by telephone. RESULTS: Age is a risk factor for Lyme disease for groups aged 10-19 years old and 50 years or older. Sex was not a risk factor. Incidence of Lyme disease in a rural setting was three times the incidence in an urban setting. Increased risk also was associated with living in single family homes, homes with yards or attached land, woods on the land, signs of tick hosts seen on the land, and homes within 100 feet of woodland. Gardening for more than four hours per week was also a risk factor, but most other outdoor activities were not. Twice as many participants took protective measures against tick bites before outdoor employment than those who merely ventured into the yard or land associated with the home. Only checking for ticks during outdoor activity and the use of repellents prior to outdoor activities outside the yard were unequivocally associated with a reduced risk of Lyme disease. CONCLUSIONS: It is important to increase public awareness about the risk of acquiring Lyme disease from ticks in the immediate environment of the home. PMID- 11889283 TI - Profile of arthritis disability. AB - Using the 1994-95 National Health Interview Supplement Disability Supplement, the authors study levels of disabilities and accommodations among US adults with arthritis disability, compared to people with disability due to other conditions. Arthritis-disabled people are defined in two ways. One definition covers a broad range of arthritis and rheumatic conditions, and the other concentrates solely on arthritis. The authors find that arthritis-disabled people have more total disabilities than other-disabled peop e. However, their disabilities are less severe, have shorter durations, and accumulate more gradually over time. Despite more disabilities, people with arthritis disability use fewer assistive and service accommodations than other-disabled people. They do use more mobility aids. Because arthritis is the leading chronic condition for middle-aged and older adults, th s profile of extensive but mild-to-moderate disability is experienced by many millions of adults. Accommodations for arthritis may also be extensive but aimed more toward self-care than toward assistive and medical services. PMID- 11889284 TI - Pregnancy feelings among adolescents awaiting pregnancy test results. AB - OBJECTIVE: The authors surveyed adolescent girls about their feelings regarding pregnancy. METHODS: A survey was administered to 117 13- to 18-year-olds who obtained pregnancy tests at nine clinics in Minneapolis and St. Paul, Minnesota, in 1998. The survey included four measures of pregnancy feelings. The authors used bivariate and multivariate logistic regression analyses to examine the associations of these measures with engagement with school, future expectations, social and environmental characteristics, and perceived partner desire for pregnancy. RESULTS: The four measures of pregnancy feelings were highly correlated (P = 0.0001). Participants reported a range of positive, negative, and ambivalent feelings on all measures. Perceived partner desire for pregnancy, limited future expectations, and lack of school engagement were significantly associated with positive pregnancy feelings for the four measures. CONCLUSIONS: Successful adolescent pregnancy prevention interventions may include the involvement of partners and key adults as well as strategies to enhance the educational or employment aspirations of girls and adolescents. PMID- 11889285 TI - Teens' images of smoking and smokers. AB - The authors used qualitative and quantitative data to identify and interpret specific images teens have about smoking and smokers. Qualitative data were collected in 1996 from 793 teenagers participating in 125 focus groups at eight different sites across the United States. Most focus groups were homogeneous with respect to gender, ethnicity, and smoking status. Ages ranged from 12 to 18 years, and about half of the participants were female. The majority of participants (62%) were white and African American, the remainder (38%) were Hispanic, American Indian, and Asian/Pacific Islander. Groups were comprised of smoking and nonsmoking teens. Focus group activities were used to elicit image related discussions about attitudes, beliefs, and perceptions of smoking. Investigators identified seven consistent and distinctive image themes: Appearance (smoking is dirty and unattractive), Activity (nonsmokers have busy, active lives), Drugs and sex (smokers are substance abusers and are sexually active), Rebellion (smokers belong to rebellious groups), Affect (smokers are depressed, angry, and stressed-out), In control (nonsmokers have self-control and are independent), and Pride (nonsmokers are proud of themselves, their families, and their heritage). A large scale, multi-site qualitative research approach can increase understanding of teen smoking. The identification of distinctive images of smoking can help researchers develop more sophisticated models of the processes of teen smoking than currently exist. PMID- 11889286 TI - Cigarette smoking and adolescents: messages they see and hear. AB - Cigarette smoking is the primary preventable cause of mortality and morbidity in the US. But in the mid-1990s, more than one-third of US teenagers were smokers, despite their awareness of the health risks and negative consequences of tobacco use. In 1996, as part of a three-year qualitative study to explore differences in adolescent smoking by gender and ethnicity, members of the Tobacco Control Network examined messages that teens receive about cigarette smoking. Consisting of 178 focus groups with 1,175 teenagers covering all levels of smoking experience, the study included teens from five ethnic groups, stratified by gender and ethnicity, from urban and rural areas across the US. The authors reviewed the sources and content of messages that teens reported were most influential in their decisions to smoke or not smoke cigarettes. Family and peers, school, television, and movies were the primary sources for both pro- and anti-smoking messages. The authors conclude that a lack of clear, consistent antismoking messages leaves teens vulnerable to the influences of pro-smoking messages from a variety of sources. Interventions need to be culture- and gender specific. Family-based interventions appear to be needed and efficacious, but resource intensive. Building self-esteem may prove to be a promising intervention. PMID- 11889287 TI - A venue-based method for sampling hard-to-reach populations. AB - Constructing scientifically sound samples of hard-to-reach populations, also known as hidden populations, is a challenge for many research projects. Traditional sample survey methods, such as random sampling from telephone or mailing lists, can yield low numbers of eligible respondents while non probability sampling introduces unknown biases. The authors describe a venue based application of time-space sampling (TSS) that addresses the challenges of accessing hard-to-reach populations. The method entails identifying days and times when the target population gathers at specific venues, constructing a sampling frame of venue, day-time units (VDTs), randomly selecting and visiting VDTs (the primary sampling units), and systematically intercepting and collecting information from consenting members of the target population. This allows researchers to construct a sample with known properties, make statistical inference to the larger population of venue visitors, and theorize about the introduction of biases that may limit generalization of results to the target population. The authors describe their use of TSS in the ongoing Community Intervention Trial for Youth (CITY) project to generate a systematic sample of young men who have sex with men. The project is an ongoing community level HIV prevention intervention trial funded by the Centers for Disease Control and Prevention. The TSS method is reproducible and can be adapted to hard-to-reach populations in other situations, environments, and cultures. PMID- 11889288 TI - Quality of data in multiethnic health surveys. AB - OBJECTIVE: There has been insufficient research on the influence of ethno cultural and language differences in public health surveys. Using data from three independent studies, the authors examine methods to assess data quality and to identify causes of problematic survey questions. METHODS: Qualitative and quantitative methods were used in this exploratory study, including secondary analyses of data from three baseline surveys (conducted in English, Spanish, Cantonese, Mandarin, and Vietnamese). Collection of additional data included interviews with investigators and interviewers; observations of item development; focus groups; think-aloud interviews; a test-retest assessment survey; and a pilot test of alternatively worded questions. RESULTS: The authors identify underlying causes for the 12 most problematic variables in three multiethnic surveys and describe them in terms of ethnic differences in reliability, validity, and cognitive processes (interpretation, memory retrieval, judgment formation, and response editing), and differences with regard to cultural appropriateness and translation problems. CONCLUSIONS: Multiple complex elements affect measurement in a multiethnic survey, many of which are neither readily observed nor understood through standard tests of data quality. Multiethnic survey questions are best evaluated using a variety of quantitative and qualitative methods that reveal different types and causes of problems. PMID- 11889290 TI - The prevention research initiative and the peer review process at CDC. PMID- 11889289 TI - A computerized tool for evaluating the effectiveness of preventive interventions. AB - In identifying appropriate strategies for effective use of preventive services for particular settings or populations, public health practitioners employ a systematic approach to evaluating the literature. Behavioral intervention studies that focus on prevention, however, pose special challenges for these traditional methods. Tools for synthesizing evidence on preventive interventions can improve public health practice. The authors developed a literature abstraction tool and a classification for preventive interventions. They incorporated the tool into a PC based relational database and user-friendly evidence reporting system, then tested the system by reviewing behavioral interventions for hypertension management. They performed a structured literature search and reviewed 100 studies on behavioral interventions for hypertension management. They abstracted information using the abstraction tool and classified important elements of interventions for comparison across studies. The authors found that many studies in their pilot project did not report sufficient information to allow for complete evaluation, comparison across studies, or replication of the intervention. They propose that studies reporting on preventive interventions should (a) categorize interventions into discrete components; (b) report sufficient participant information; and (c) report characteristics such as intervention leaders, timing, and setting so that public health professionals can compare and select the most appropriate interventions. PMID- 11889291 TI - Predictive value of antithrombin III and serum C-reactive protein concentration in critically ill patients with suspected sepsis. AB - OBJECTIVE: To evaluate at admission the performance of serum antithrombin III, serum C-reactive protein, white blood cell and platelet counts, and thromboplastin time values in prediction of hospital mortality rates in critically ill patients with suspected sepsis. DESIGN: Prospective, cohort study. SETTING: University hospital medical-surgical intensive care unit. PATIENTS: One hundred eight consecutive critically ill patients with suspected sepsis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The outcome measure was hospital mortality rate. Hospital survivors (n = 66) and nonsurvivors (n = 42) differed statistically significantly in admission antithrombin III activity (percentage of normal): survivors' median 66% (interquartile range, 48% to 82%) vs. nonsurvivors' median 46% (37% to 65%, p =.0002 by Mann-Whitney test). Analysis revealed similarly statistically significant differences between survivors and nonsurvivors in admission platelet count, admission thromboplastin time, day 1 Logistic Organ Dysfunction score, and Acute Physiology and Chronic Health Evaluation III score, but not in serum C-reactive protein concentrations or in white blood cells. However, the areas under the receiver operating curves (AUC) showed significantly worse discriminative power for admission antithrombin III concentration (AUC, 0.71; SE, 0.05), platelet count (AUC, 0.67; SE, 0.05), thromboplastin time (AUC, 0.65; SE, 0.05), C-reactive protein concentration (AUC, 0.60; SE, 0.05), and white blood cell count (AUC, 0.53; SE, 0.06) than did the day 1 Logistic Organ Dysfunction score (AUC, 0.82; SE, 0.04) and the Acute Physiology and Chronic Health Evaluation III score (AUC, 0.84; SE, 0.04). Multivariate logistic regression analysis revealed that only the Acute Physiology and Chronic Health Evaluation III score was independently associated with hospital mortality rate. CONCLUSIONS: Admission antithrombin III concentrations, but not C-reactive protein concentrations, differ significantly between hospital survivors and nonsurvivors among critically ill patients with septic infection. However, in prediction of hospital mortality rate, the discriminative power of admission antithrombin III concentration is poor, as judged by analysis of areas under the receiver operating curves, and is not independently associated with hospital mortality rate. PMID- 11889292 TI - Endotoxin-induced mitochondrial damage correlates with impaired respiratory activity. AB - OBJECTIVE: This study was designed to determine whether mitochondrial function in a systemic organ is acutely impaired in a resuscitated model of sepsis (endotoxemia, lipopolysaccharide) and the relationship, if any, between this impairment and the extent of mitochondrial ultrastructural damage that occurs. DESIGN: Perspective, controlled laboratory investigation. SETTING: Animal laboratory in a university research institute. SUBJECTS: Adult male cats. INTERVENTIONS: A well-established feline model of acute endotoxemia was used wherein measures were taken to minimize tissue hypoxia. After lipopolysaccharide (3 mg/kg intravenously, n = 9) or isotonic saline vehicle (control, n = 5) administration, liver samples were obtained at 4 hrs posttreatment, and mitochondrial ultrastructure and respiratory function were assessed. Mitochondrial ultrastructural injury was graded on a scale of 0 (no injury) to 5 (severe injury), and mitochondrial respiration was evaluated by using standard techniques. MEASUREMENTS AND MAIN RESULTS: Significant mitochondrial injury was apparent by 4 hrs, but only in the lipopolysaccharide-treated group (2.5 +/- 0.2 vs. 1.3 +/- 0.2, p <.001) and despite maintenance of tissue oxygen availability. In addition, lipopolysaccharide treatment reduced the rate of state 3 (adenosine 5'-diphosphate-dependent) respiration, especially at complex IV (40% inhibition), and increased the rate of state 4 (adenosine 5'-diphosphate-independent) respiration, reflecting partial uncoupling of mitochondrial oxidative phosphorylation. Finally, a significant correlation was demonstrated between the severity of ultrastructural injury and the magnitude of mitochondrial respiratory dysfunction after lipopolysaccharide treatment and despite resuscitation efforts. CONCLUSION: Mitochondrial function is significantly impaired during acute sepsis, and this impairment is strongly associated with the extent of mitochondrial ultrastructural abnormalities present in the tissues. These findings in conjunction with those previously shown suggest that mitochondrial functional impairment may contribute to the pathogenesis of altered oxygen metabolism in systemic organs during sepsis. PMID- 11889294 TI - Mechanical ventilation with permissive hypercapnia increases intrapulmonary shunt in septic and nonseptic patients with acute respiratory distress syndrome. AB - OBJECTIVE: To compare the effects of conventional mechanical ventilation with low volume, pressure-limited ventilation (LVPLV) and permissive hypercapnia on ventilation-perfusion (V/Q) distributions in patients with acute respiratory distress syndrome. We hypothesized that the advantageous cardiopulmonary effects of LVPLV would be greater in patients with sepsis than in those without sepsis. PATIENTS AND INTERVENTIONS: Twenty-two patients with acute respiratory distress syndrome were studied (group 1: 12 patients with hyperdynamic sepsis; group 2: 10 nonseptic patients). Intrapulmonary shunt (Qsp/Qt) (percentage of cardiac output), perfusion of "low" V/Q areas (percentage of cardiac output), ventilation of "high" V/Q areas (percentage of total ventilation [VE]), and deadspace ventilation (percentage of VE) were calculated from the retention/excretion data of six inert gases. Data were obtained during conventional mechanical ventilation and during LVPLV. MEASUREMENTS AND MAIN RESULTS: In group 1, LVPLV increased PaCO(0)rom 38 +/- 6 torr (5.1 +/- 0.8 kPa) to 61 +/- 12 torr (8.1 +/- 1.6 kPa). Qsp/Qt increased from 28 +/- 16% to 36 +/- 17%, whereas Pao2 (84 +/- 15 torr [11.1 +/- 2.0 kPa] vs. 86 +/- 21 torr [11.5 +/- 2.8 kPa]) and Qt (10.6 +/- 2.3 vs. 11.5 +/- 2.5 L x -1) remained unchanged and PVO(2) (40 +/- 4 [5.3 +/- 0.5 kPa] vs. 49 +/- 6 torr [6.5 +/- 0.3]) increased. In group 2, LVPLV increased PaCO(2) from 38 +/- 6 torr (5.1 +/- 0.8 kPa) to 63 +/- 11 torr (8.4 +/- 1.5 kPa). For Qsp/Qt (24 +/- 9% to 34 +/- 16%), the increase was not significant, whereas Qt (7.4 +/- 1.8 vs. 10.2 +/- 2.2 L x -1), PVO(2)(38 +/- 4 torr [5.1 +/- 0.5 kPa] vs. 50 +/- 6 mm Hg [6.7 +/- 0.8 kPa]), and PaO(2) (89 +/- 16 torr [11.9 +/- 2.1 kPa] vs. 98 +/- 19 torr [13.1 +/- 2.5 kPa]) increased. In both groups, the scatter of perfusion distribution (log SDQ) was greater than expected for normal subjects but was not different between the groups or altered by the treatments. CONCLUSIONS: In patients with acute respiratory distress syndrome, LVPLV with permissive hypercapnia, tended to increase Qsp/Qt, without a concomitant decrease of PaO(2). This occurs because, although atelectasis and increased shunt result from the low ventilatory volume, the effects on PaO(2) are offset by increased PVO(2) resulting from the hypercapnic stimulation of cardiac output. This result was independent of the presence or absence of sepsis. PMID- 11889295 TI - Education, ethics, and end-of-life decisions in the intensive care unit. AB - OBJECTIVE: To examine the influence of education and clinical experience on residents' attitudes toward withdrawal of life support. DESIGN: Self-administered survey. SETTING: Four Canadian teaching hospitals. SUBJECTS: Residents rotating through four intensive care units. MEASUREMENTS AND MAIN RESULTS: The survey examined ethics education and experience regarding end-of-life care, importance of factors influencing withdrawal of life support, confidence in decisions, and recommendations for enhancing end-of-life education. The response rate was 83.9% (52 of 62). A minority of residents reported an appropriate amount of formal teaching on ethical principles (17.3%), patient-centered education (28.8%), and informal discussion (28.8%) before their intensive care unit rotation. During their rotation, most residents cared for patients in whom withdrawal of life support was considered. Although they usually attended family meetings, residents were never (34.6%) or rarely (42.3%) the primary discussant. Before the intensive care unit rotation, confidence in withdrawal decisions was related to male sex (p =.001) and previous patient-centered ethics education (p =.02). At the end of the intensive care unit rotation, only resident involvement in family meetings (p =.02) and being the primary discussant at such meetings (p =.01) were associated with confidence. After we adjusted for pre-rotation confidence in withdrawal of life support decision-making, the only predictor of post-rotation confidence was family meeting involvement (p <.001). Residents recommended more patient-centered discussion, observation of attending physicians discussing end-of-life issues, and opportunity to lead family meetings. CONCLUSIONS: Experiential, case-based, patient-centered curricula are associated with resident confidence in withdrawal of life support decisions in the intensive care unit. PMID- 11889297 TI - Head position for facilitating the superior vena caval placement of catheters during right subclavian approach in children. AB - OBJECTIVE: To evaluate the effect of head position as a method to facilitate the superior vena caval placement of catheters during right subclavian catheterization in children. DESIGN: Prospective, randomized, clinical trial. SETTING: Department of anesthesiology, university hospital. PATIENTS: One hundred sixty-eight pediatric patients, aged <8 yrs, undergoing simple cardiac surgery or pediatric general surgery. INTERVENTIONS: At operation, the patients were assigned by the stratified randomization for age to one of the four groups (n = 42 each): when the patients turned their heads away from the puncture side, this was away-turning group; when turned toward the puncture side, toward-turning group; when lateral-flexed (tilted) away from the puncture side, away-lateral flexion group; and when lateral-flexed toward the puncture side, toward-lateral flexion group. Each group was divided into two subgroups depending on the age: infant (n = 24 each) and young children (>12 months; n = 18 each). MEASUREMENTS AND MAIN RESULTS: Right infraclavicular subclavian catheterization, using the Seldinger technique, was attempted. After catheterization, a simple chest radiograph was used to identify the location of catheter tip. There was no difference in age and body weight between the groups. Only in infants was the successful placement rate of toward-lateral-flexion group (92%) higher than that of the other three groups (54% [away-lateral-flexion], 63% [away-turning], or 54% [toward-turning]), and there was no difference among the others. CONCLUSION: In infants, tilting the head toward the catheterization side can reduce the incidence of catheter malposition during the right subclavian approach. PMID- 11889299 TI - Brain death assessment using instant spectral analysis of heart rate variability. AB - OBJECTIVE: Confirmation of brain death requires an urgent diagnosis to allow rapid vital organ removal for transplantation. Evaluation of forebrain functions is commonly performed through electroencephalogram. Nevertheless, there are, for the moment, no methods that allow for an instantaneous evaluation of brainstem functions. During acute brain injury, heart rate variability is an independent neurologic prognosis indicator resulting from a close relationship between brain stem and cardiac autonomic nervous system. This study aims to evaluate a new heart rate variability spectral analysis method, on a beat-to-beat basis, continuously over the time, during brain death. DESIGN: Prospective, nonrandomized, observational study. SETTING: Intensive care unit. SUBJECTS: Ten patients (age range 25-64 yrs, mean age 41 yrs) with acute brain injury leading to brain death. INTERVENTION: No intervention beyond standard of care MEASUREMENTS AND MAIN RESULTS: Heart rate, arterial blood pressure, heart rate variability in time and frequency domains method, which included calculation of the instant center frequency of spectrum. Brain death was associated with tachycardia (R-R interval 703 +/- 69 vs. 551 +/- 34 msec, p <.05), dramatic reduction of the global spectral power (44.919 +/- 31.511 vs. 3.204 +/- 1.469 msec(2), p <.05), and an abrupt shift of instant center frequency to a higher frequency range (0.17 +/- 0.01 vs. 0.26 +/- 0.03 Hz, p <.05). CONCLUSIONS: Such a method allows an instant, noninvasive determination of brainstem death based on a time and frequency domain analysis of heart rate variability. PMID- 11889298 TI - Fluid resuscitation and hyperchloremic acidosis in experimental sepsis: improved short-term survival and acid-base balance with Hextend compared with saline. AB - OBJECTIVE: To compare resuscitation with 0.9% saline with Hextend, a synthetic colloid in a balanced electrolyte solution, in terms of acid-base status and survival time in an experimental model of septic shock in the rat. DESIGN: Randomized, open-label, controlled experiment. SETTING: University research laboratory. SUBJECTS: Sixty adult, male Sprague-Dawley rats. INTERVENTION: Animals were studied for 12 hrs after intravenous infusion of Escherichia coli endotoxin (20 mg/kg). Animals were volume resuscitated to maintain a mean arterial pressure >60 mm Hg using either 0.9% saline (n = 25), Hextend (n = 25), or lactated Ringer's (n = 10). MEASUREMENTS: Arterial blood gases and electrolytes were measured before and after resuscitation (0, 180, 360, and 540 mins after endotoxin infusion). Survival time was measured, up to 12 hrs. RESULTS: Mean survival time among animals treated with saline or Ringer's was 45% less compared with Hextend-treated animals: 391 +/- 151 mins and 362 +/- 94 mins vs. 567 +/- 140 mins, respectively, p <.0001. Overall survival (at 12 hrs) was 0% with saline or Ringer's vs. 20% with Hextend, p =.05. After resuscitation with saline, arterial standard base excess and plasma apparent strong ion difference were both significantly lower (-19.3 +/- 5.2 vs. -12.1 +/- 5.7, p <.001, and 23.0 +/- 6.2 vs. 30.3 +/- 2.9, p <.0001, respectively) and plasma Cl(-) was significantly higher (123 +/- 7 vs. 115 +/- 3 mmol/L, p <.0001) compared with Hextend. Resuscitation with Ringer's solution resulted in a standard base excess, and Cl(-) between that of saline and Hextend (-15.4 +/- 3.1, and 117 +/- 3, respectively). CONCLUSION: Compared with 0.9% saline, volume resuscitation with Hextend was associated with less metabolic acidosis and longer survival in this experimental animal model of septic shock. PMID- 11889301 TI - Impairment of polymorphonuclear neutrophil functions precedes nosocomial infections in critically ill patients. AB - OBJECTIVE: A postinjury immunodepression involving neutrophil functions has been described in critically ill patients. The aim of this prospective study was to search for a relationship between an impairment of neutrophil functions and the subsequent development of nosocomial infection. DESIGN: Twenty-one severely ill (simplified acute physiology score II >20 on admission), nonimmunosuppressed patients who were receiving no antibiotics active against methicillin-resistant Staphylococcus aureus and highly resistant Pseudomonas aeruginosa were included. Twelve healthy subjects constituted a control group. MEASUREMENTS: Neutrophil functions (phagocytosis and bactericidal activity toward S. aureus and P. aeruginosa in homologous plasma, reactive oxygen species secretion) were studied at day 4 +/- 1 after admission, and occurrence of nosocomial infection was prospectively recorded over the following 5 days. Interleukin-10 concentration was assessed by enzyme-linked immunosorbent assay. Results are expressed as median (25th-75th percentiles). MAIN RESULTS: Six out of the 21 patients acquired a nosocomial infection during the 5 days after blood sampling (infected group). Compared with the patients who did not acquire nosocomial infection (noninfected group, n = 15), the neutrophils of the infected group demonstrated a higher percentage of intracellular bacterial survival (17% [2% to 67%] vs. infected: 62% [22% to 100%], p <.05), leading to an impairment of S. aureus killing in homologous plasma (killed bacteria: 4.93 log(10) colony forming units/mL [4.24 5.29] vs. infected: 3.62 log(10) colony forming units/mL [0.00-4.58], p <.05). Interleukin-10 plasma concentration was higher in infected patients (78 pg/mL [60 83]) compared with noninfected patients (22 pg/mL [14-58], p <.05). By contrast, P. aeruginosa killing was similar in patients whether or not they acquired a nosocomial infection. CONCLUSION: A decrease in S. aureus killing capabilities of neutrophils can be evidenced within the days before occurrence of a nosocomial infection. PMID- 11889300 TI - Effect of enriched thioglycolate on direct examination of respiratory specimens and guiding initial empirical therapy in intubated patients with pneumonia: a prospective, randomized study. AB - OBJECTIVE: To evaluate the use of enriched thioglycolate as a transport medium of protected specimen brush samples, in particular its value in direct examination of respiratory specimens and in guiding initial antibiotic prescription. DESIGN: Prospective, randomized, pilot study. SETTING: Medical-surgical teaching intensive care unit. SUBJECTS: Thirty adults with suspected ventilator-associated pneumonia. INTERVENTION: Bronchoscopy was performed by using standard techniques, and two consecutive protected specimen brush samples were taken. Transport medium consisted of 1 mL of sterile saline or thioglycolate. Gram stains were performed in all samples. Randomization was used to select which transport medium was used first. Each patient served as his or her own control. The laboratory was blind to the choice. MEASUREMENTS AND MAIN RESULTS: Causative agents were cultured in 16 episodes, and no significant differences were observed when the transport media were compared. Anaerobes were identified in only one episode. Direct staining in thioglycolate samples anticipated the presumptive diagnosis (in presence of false negative cultures) in three additional patients in whom prior antibiotic therapy was prescribed; this was also the case in one patient with saline solution. The etiology was anticipated by direct Gram staining and permitted a more targeted initial empirical treatment in 75% of samples transported in thioglycolate, compared with only 37.5% of samples transported in saline solution (p <.05). CONCLUSION: When protected specimen brush samples are obtained, thioglycolate may contribute to early identification of the pathogen and may guide the initial empirical therapy. PMID- 11889302 TI - Noninvasive proportional assist ventilation compared with noninvasive pressure support ventilation in hypercapnic acute respiratory failure. AB - OBJECTIVES: To compare short-term administration of noninvasive proportional assist ventilation (NIV-PAV) and pressure support ventilation (NIV-PSV). DESIGN: Prospective, crossover, randomized study. SETTING: Medicosurgical intensive care unit in a nonteaching hospital. PATIENTS: Twelve chronic obstructive pulmonary disease patients admitted for hypercapnic acute respiratory failure. INTERVENTION: NIV-PSV and NIV-PAV given in a randomized order after baseline evaluation in continuous positive airway pressure. Using a flow-triggering ventilator, NIV-PAV was adjusted using the runaway method and compared with NIV PSV at similar peak inspiratory airway pressure. MEASUREMENTS AND MAIN RESULTS: Flow, airway pressure, and changes in esophageal pressure were measured and the tidal volume, the patient's inspiratory work of breathing, and the esophageal pressure--time product were calculated. Arterial pH and PaCO(2) were measured and breathing comfort was assessed using a visual analogic scale. Peak inspiratory airway pressure (17 +/- 3 cm H(2)O) and tidal volume were similarly increased with the two modalities with no change in respiratory rate. The change in esophageal pressure was similarly decreased (from 20 +/- 8 cm H(2)O in continuous positive airway pressure to 12 +/- 7 in NIV-PSV and 10 +/- 5 cm H(2)O in NIV-PAV) as well as inspiratory muscle effort indexes. Arterial pH and PaCO(2) were similarly improved. Breathing comfort was significantly improved in NIV-PAV (+38 +/- 38%) but not in NIV-PSV (+11 +/- 23%). The tidal volume was more variable in NIV-PAV (89 +/- 18%) than in NIV-PSV (15 +/- 8%) and changes in tidal volume variability were significantly correlated (p =.02) with changes in breathing comfort. CONCLUSIONS: In chronic obstructive pulmonary disease patients with hypercapnic acute respiratory failure, NIV-PAV was able to unload inspiratory muscles similarly to NIV-PSV but may be more comfortable than NIV-PSV. PMID- 11889305 TI - Discriminative power on mortality of a modified Sequential Organ Failure Assessment score for complete automatic computation in an operative intensive care unit. AB - OBJECTIVE: To evaluate the discriminative power on mortality of a modified Sequential Organ Failure Assessment (SOFA) score and derived measures (maximum SOFA, total maximum SOFA, and delta SOFA) for complete automatic computation in an operative intensive care unit (ICU). DESIGN: Retrospective study. SETTING: Operative ICU of the Department of Anesthesiology and Intensive Care Medicine. PATIENTS: Patients admitted to the ICU from April 1, 1999, to March 31, 2000 (n = 524). Data from patients under the age of 18 yrs and patients who stayed <24 hrs were excluded. In the case of patient readmittance, only data from the patient's last stay was included in the study. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The main outcome measure was survival status at ICU discharge. Based on Structured Query Language (SQL) scripts, a modified SOFA score for all patients who stayed in the ICU in 1 yr was calculated for each day in the ICU. Only routine data were used, which were supplied by the patient data management system. Score evaluation was modified in registering unavailable data as being not pathologic and in using a surrogate of the Glasgow Coma Scale. During the first 24 hrs, 459 survivors had an average SOFA score of 4.5 +/- 2.1, whereas the 65 deceased patients averaged 7.6 +/- 2.9 points. The area under the receiver operating characteristic (ROC) curve was 0.799 and significantly >0.5 (p <.01). A confidence interval (CI) of 95% covers the area (0.739-0.858). The maximum SOFA presented an area under the ROC of 0.922 (CI: 0.879-0.966), the total maximum SOFA of 0.921 (CI: 0.882-0.960), and the delta SOFA of 0.828 (CI: 0.763-0.893). CONCLUSION: Despite a number of differences between completely automated data sampling of SOFA score values and manual evaluation, the technique used in this study seems to be suitable for prognosis of the mortality rate during a patient's stay at an operative ICU. PMID- 11889304 TI - Predictors and impact of atrial fibrillation after isolated coronary artery bypass grafting. AB - OBJECTIVE: Although an extensive number of studies have attempted to identify predictors of new-onset atrial fibrillation (AFIB) after coronary artery bypass grafting (CABG), a strong predictive model does not exist. Prior studies have included patients recruited from multiple centers with variant AFIB prevalence rates and those who underwent CABG in combination with other surgical procedures. Also, most studies have focused on pre- and perioperative characteristics, with less attention given to the initial postoperative period. The purpose of this study was to comprehensively examine pre-, peri-, and postoperative characteristics that might predict new-onset AFIB in a large sample of patients undergoing isolated CABG in a single medical center, utilizing data readily available to clinicians in electronic data repositories. In addition, length of stay and selected postoperative complications and disposition were compared in patients with AFIB and no AFIB. DESIGN: Retrospective, comparative survey. SETTING: University-affiliated tertiary care hospital. PATIENTS: Patients with new-onset AFIB who underwent isolated standard CABG or minimally invasive direct vision coronary artery bypass were identified from an electronic clinical data repository. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The prevalence of AFIB in the total sample (n = 814) was 31.9%. Predictors of AFIB included age (p =.0004), number of vessels bypassed (p =.013), vessel location (diagonal [p <.003] or posterior descending artery [p <.001]), and net fluid balance on the operative day (p =.015). Forward stepwise regression analysis produced a model that correctly predicted AFIB in only 24% of cases, with age (14%) and body surface area (9%) providing the most prediction. The incidence of embolic stroke was higher in AFIB (n = 8) vs. no AFIB (n = 4) patients, but stroke preceded AFIB onset in seven of eight cases. Subjects with AFIB had a longer stay (p =.0004), more intensive care unit readmissions (p =.0004), and required more assistance at hospital discharge (p =.017). CONCLUSIONS: Despite attempts to examine comprehensively predictors of new-onset AFIB, we were unable to identify a robust predictive model. Our findings, in combination with prior work, imply that it may not be feasible to predict the development of new-onset AFIB after CABG using data readily available to the bedside clinician. In this sample, stroke was uncommon and, when it occurred, preceded AFIB in all but one case. As anticipated, AFIB increased length of stay, and patients with this complication required more assistance at discharge. PMID- 11889306 TI - Effect of age and birth weight on indomethacin pharmacodynamics in neonates treated for patent ductus arteriosus. AB - OBJECTIVES: To determine patent ductus arteriosus (PDA) closure rates, and indomethacin (INDO) toxicity rates in neonates dosed with INDO using an individualized pharmacokinetic/pharmacodynamic (PK/PD) dosing approach. In addition, develop PD curves evaluating dose-response and concentration-response relationships for closure and renal toxicity, especially in select subgroups historically known as "poor responders" (<1000 g and > or = 10 days postnatal age). DESIGN: Prospective, cohort study. SETTING: Level III neonatal intensive care unit. SUBJECTS: One hundred thirty-nine patients receiving 151 courses of INDO for PDA closure were evaluated. INTERVENTIONS: Patients initially received 0.25 mg/kg of INDO, followed immediately by 1 mg/kg of furosemide. INDO concentrations were obtained 2 hrs and 8 hrs after the dose and were assayed using high-performance liquid chromatography. Individualized PK parameters were calculated with subsequent INDO dosing based on the individualized PK variables to increase trough serum concentrations by 0.3-0.5 mg/L. MEASUREMENTS AND MAIN RESULTS: Ductal closure was successful in 127 patients (91%). Renal toxicity occurred in 21 (15%) patients and was temporary and reversible. No significant differences in response rates based on treatment weight or postnatal age were observed. PD curves were similar for neonates <1000 g vs. > or = 1000 g. PD curves were also similar for neonates with postnatal age <10 days vs. > or = 10 days. Statistically significant differences were noted between neonates categorized for postnatal age <10 days vs. > or = 10 days in total days of therapy (1.8 vs. 2.3 days), total number of doses required to close PDA (3.5 vs. 5.6 doses), critical INDO dose (0.9 vs. 1.4 mg/kg), critical INDO concentration (1.9 vs. 1.4 mg/L), and critical dose/critical concentration ratio (0.52 vs. 2.2). CONCLUSIONS: These findings support the hypothesis that the poor PDA closure rates with INDO for neonates >10 days postnatal age are the result of pharmacokinetic differences only and that weight does not impact response rates. Individualized pharmacokinetic/pharmacodynamic dosing of INDO continues to achieve higher closure rate than current dosing standards. Patients historically known as poor responders significantly benefit from this dosing approach. PMID- 11889308 TI - Evaluation of endotoxin release and cytokine production induced by antibiotics in patients with Gram-negative nosocomial pneumonia. AB - OBJECTIVE: To determine the plasma concentrations of lipopolysaccharide, tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6 in a homogeneous group of septic patients and to evaluate the effect of antibiotic treatment, imipenem or ceftazidime, on the release of lipopolysaccharide and cytokines. DESIGN: Prospective, randomized study. SETTING: Sixteen-bed multidisciplinary intensive care unit. PATIENTS: Twenty-four septic patients with documented Gram negative nosocomial pneumonia. Controls were 20 patients admitted without sepsis and 20 healthy volunteers. INTERVENTIONS: Septic patients were randomized between imipenem and ceftazidime. Blood samples were collected before (0 hrs) and after (4 and 12 hrs) antibiotic treatment. Concentrations of lipopolysaccharide were measured by using the limulus assay, and cytokine concentrations were measured by enzyme-linked immunosorbent assay. Statistical analyses were performed by Kruskal Wallis test, Mann-Whitney U test, and Student's t-test. MEASUREMENTS AND MAIN RESULTS: The mean age was 48.5 +/- 19.5. The mean Acute Physiology and Chronic Health Evaluation II score was 18.4 +/- 4.5. Overall mortality rate was 45.4%. All septic patients showed significant higher concentrations of lipopolysaccharide (p <.001), tumor necrosis factor-alpha (p <.04), and interleukin-6 (p <.001) than the controls, but interleukin-1 beta was never detected. We did not find statistically significant changes in lipopolysaccharide or cytokine plasma concentrations over time within any of the two arms of the study (ceftazidime vs. imipenem). There were no statistically significant differences in lipopolysaccharide and interleukin-6 plasma concentrations between the two antibiotic treatments. Although tumor necrosis factor-alpha plasma concentrations were significantly higher in the group treated with ceftazidime compared with the group treated with imipenem at the baseline and 4 hrs later, these differences were not statistically significant after 12 hrs of initiation of both treatments. CONCLUSIONS: Patients with Gram-negative nosocomial pneumonia have high plasma concentrations of lipopolysaccharide, interleukin-6, and tumor necrosis factor-alpha, but the antibiotic therapy evaluated did not significantly modify these concentrations. PMID- 11889311 TI - Evaluation of left ventricular filling pressure by transthoracic Doppler echocardiography in the intensive care unit. AB - OBJECTIVE: To determine whether Doppler transmitral and pulmonary venous flow pattern is related to left ventricular filling pressures in critically ill patients. DESIGN: Prospective clinical investigation. SETTING: Medical intensive care unit of a university hospital. PATIENTS: Fifty-four mechanically ventilated patients (age, 63 +/- 16 yrs) were investigated via transthoracic echocardiography and Doppler. Main diagnoses were pneumonia (31%), acute exacerbation of chronic obstructive pulmonary disease (24%), congestive heart failure (11%), and poisoning (11%). INTERVENTIONS: Doppler examinations were performed simultaneously with measurements of pulmonary artery occlusion pressure via a right heart catheter. MEASUREMENTS AND MAIN RESULTS: Pulmonary artery occlusion pressure correlated with transmitral peak E-wave velocity (r =.46) and E/A ratio (r =.55). Pulmonary artery occlusion pressure inversely correlated with deceleration time of the transmitral E-wave (r = -.52), pulmonary venous peak S wave velocity (r = -.37), and systolic fraction of the pulmonary forward flow (r = -.56). An E/A ratio >2 predicted a pulmonary artery occlusion pressure >18 mm Hg with a positive predictive value of 100%. A duration of pulmonary venous A wave reversal flow exceeding the duration of the transmitral A-wave forward flow predicted a pulmonary artery occlusion pressure >15 mm Hg with a positive predictive value of 83%. A systolic fraction of the pulmonary venous forward flow <0.4 predicted a pulmonary artery occlusion pressure >12 mm Hg with a positive predictive value of 100%. CONCLUSION: Transmitral and pulmonary venous flow patterns measured by transthoracic Doppler echocardiography can be used to estimate the left ventricular filling pressure in critically ill patients. PMID- 11889309 TI - Energy balance in the intubated human airway is an indicator of optimal gas conditioning. AB - OBJECTIVE: The optimal level of inspired heat and humidity for patients receiving long-term mechanical ventilation is still the subject of debate. Many laboratory studies examining surrogate markers for optimal humidity suggest that inspired gas should be at body temperature and fully saturated. The aim of this study was to determine the inspired gas condition that was thermodynamically neutral to the airway of intubated patients, and also examine the contribution of the endotracheal tube to airway heat and water balance. DESIGN: Prospective, block randomized, observational study. SETTING: General adult intensive care unit of a metropolitan teaching hospital. PATIENTS: Ten adult patients requiring intermittent positive pressure ventilation for nonpulmonary reasons. INTERVENTIONS: Each patient was given four different gas conditions--30 degrees C, 30 mg/L; 34 degrees C, 38 mg/L; 37 degrees C, 44 mg/L; and 40 degrees C, 50 mg/L--to breathe in random order. MEASUREMENTS AND MAIN RESULTS: Inspired and expired gas temperature and humidity, and the temperature gradient down the endotracheal tube, were measured and the inspired gas condition that gave thermodynamic neutrality was determined. This was found to be gas at body temperature, saturated. Airway workload and airway water loss increased linearly as the inspired gas departed from this condition, at approximately 1.4 kJ/hr/ degrees C and 0.5 mL/hr/ degrees C, respectively. The endotracheal tube contributed very little to heat and water exchange. CONCLUSIONS: Inspired gas at body temperature and saturated is thermodynamically neutral to the intubated airway, and thus may be considered the optimal condition for ventilation lasting more than a few hours. PMID- 11889312 TI - Diagnostic and therapeutic management of nosocomial pneumonia in surgical patients: results of the Eole study. AB - OBJECTIVE: To assess clinical, microbiological, and therapeutic features of nosocomial pneumonias in surgical patients. DESIGN: Prospective (October 1997 through May 1998), consecutive case series analysis of patients suspected of having pneumonia during the fortnight after a surgical procedure or trauma and receiving antibiotic therapy prescribed by the attending physician for this diagnosis. SETTING: A total of 230 study centers in teaching (n = 66) and nonteaching hospitals (n = 164) (surgical wards and intensive care units). PATIENTS: A total of 837 evaluable patients (mean age 61 +/- 18 yrs) including 629 intensive care unit patients. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: The diagnostic and therapeutic procedures followed were based on guidelines. Antibiotics and any changes of therapy and duration of treatment were decided by the attending physician. The charts were reviewed by a panel of experts that classified the cases according to clinical, radiologic, and microbiological criteria (when available). The efficacy of treatment was evaluated over a 30-day period following the index episode. The patients were classified into three groups: definite pneumonia (n = 261), possible pneumonia (n = 392), or low-probability pneumonia (n = 184). Ventilator-acquired pneumonia was reported in 303 patients. Early onset pneumonia was reported in 512 cases. Microbiological sampling was performed in 718 patients, by bronchoscopy in 367 cases, recovering 450 organisms in 328 patients, including 94 polymicrobial specimens. High proportions of Gram-negative bacteria and staphylococci were cultured, even in early onset pneumonias. Antibiotic therapy was administered for 13 +/- 4 days, using monotherapy in 254 cases. Changes in the initial antibiotic therapy (135 monotherapies) were decided in 517 patients (including clinical failure or persistent infection, n = 171; organisms resistant to initial therapy, n = 177; pulmonary superinfection, n = 68). Death occurred in 180 patients, related to pneumonia in 53 cases. CONCLUSIONS: Nosocomial pneumonias in surgical patients are characterized by high frequency of early onset pneumonia, high proportion of nosocomial organisms even in these early onset pneumonias, and moderate mortality rate. PMID- 11889314 TI - Typhoid, hepatitis E, or typhoid and hepatitis E: the cause of fulminant hepatic failure--a diagnostic dilemma. AB - OBJECTIVE: To report a case of hepatitis E-induced fulminant hepatic failure associated with typhoid fever, diagnosed with the Widal test. DESIGN: Case report. SETTING: Eight-bed medical/surgical intensive care unit of a university hospital. PATIENT: A 15-yr-old, 50-kg male with grade IV hepatic encephalopathy was admitted to the intensive care unit for ventilatory support. On admission to the intensive care unit he had had fever associated with loss of appetite and nausea for 15 days, jaundice for 4 days, and altered sensorium for 2 days. INTERVENTION: He was intubated and kept on elective ventilation. Tracheal aspirate, blood, urine, and stool were sterile. Anti-coma measures were instituted in the form of 20 degrees head elevation; mannitol, lactulose, and ampicillin through a nasogastric tube; and bowel wash. The mainstay of fluid therapy was 20% dextrose. Viral marker was positive for hepatitis E. He showed a favorable recovery but continued to have high-grade fever (39-40 degrees C). On investigation, peripheral blood smear was negative for malarial parasite, and Widal was positive. Fever responded to treatment with Ceftazidime. RESULT: The patient recovered with anti-coma and anti-typhoid therapy. CONCLUSION: In viral hepatitis, fever is usually present in the prodromal phase but subsides before appearance of the icteric phase. In endemic areas, if fever is present in the icteric phase of hepatitis, typhoid also should be considered in the differential diagnosis of fever, even in the absence of positive cultures for Salmonella typhi. The Widal test may be helpful in reaching a diagnosis. PMID- 11889315 TI - Small intestine intramucosal PCO(2) and microvascular blood flow during hypoxic and ischemic hypoxia. AB - OBJECTIVE: To determine whether small intestine intramucosal PCO(2) and mucosal blood flow changes would be different between ischemic and hypoxic hypoxia. DESIGN: Randomized animal experiment. SETTING: Research laboratory. SUBJECTS: Anesthetized, mechanically ventilated, and surgically instrumented pigs. INTERVENTIONS: Systemic oxygen delivery was lowered in a stepwise manner to decrease it beyond critical oxygen delivery by lowering either FIO(2) or blood volume. MEASUREMENTS AND MAIN RESULTS: In hypoxic hypoxia pigs (n = 6), arterial oxygen concentration and oxygen delivery decreases were achieved by progressively reducing arterial PO(2) while cardiac index remained unchanged. In ischemic hypoxia pigs (n = 5), oxygen delivery reduction was achieved by progressively reducing cardiac index while arterial PO(2) remained unchanged. In control pigs, oxygen delivery remained unchanged. The lowest oxygen delivery measured in both hypoxia and ischemia experiments was 3.60 +/- 0.26 vs. 2.93 +/- 0.77 mL x kg(-1) x min(-1), respectively (p =.23). At the lowest oxygen delivery level, differences between ischemic hypoxia and hypoxic hypoxia experiments were observed for arterial lactate concentration (468 +/- 308 vs. 1070 +/- 218 mmol/L, respectively; p =.03), mixed venous arterial PCO(2) difference (10 +/- 7 vs. 4 +/ 2 torr, respectively; p =.04), and small intestine mucosal blood flow (6.2 +/- 2.1 vs. 15.7 +/- 7.4 perfusion units, respectively; p =.02). Small intestine intramucosal-arterial difference was higher in ischemic hypoxia than in hypoxic hypoxia (52 +/- 15 vs. 31 +/- 12 torr, respectively; p =.03). CONCLUSION: Small intestine intramucosal PCO(2) increases may indicate systemic oxygen uptake supply limitation in ischemic and hypoxic hypoxia related to conditions of mucosal flow stagnation and CO(2) generation. PMID- 11889316 TI - Hemodynamic responses to fluid resuscitation after blunt trauma. AB - OBJECTIVES: To determine the acute circulatory effects of an open fracture and the circulatory effects of different fluid resuscitation strategies after such a fracture. DESIGN: Randomized controlled laboratory study. SETTING: Medical Research Council laboratory, university medical school. SUBJECTS: Four groups of ten immature female Large-White pigs (16.5-27.0 kg): those receiving limited resuscitation, moderate resuscitation, and normal resuscitation, as well as surgical controls. INTERVENTIONS: After induction and intubation with 3% halothane, anesthesia was maintained with intravenous Saffan. Cannulae were placed in the left external jugular vein and both axillary arteries. A pulmonary artery flotation catheter was introduced through the jugular cannula. The left femur was exposed for fracture with a captive bolt. Resuscitation was with 0.9% saline; the moderate resuscitation group received less than the normal resuscitation group (840 +/- 67 mL vs. 1873 +/- 96 mL), whereas the surgical control and limited resuscitation groups received the lowest volumes (466 +/- 10 mL and 452 +/- 19 mL, respectively). MEASUREMENTS AND MAIN RESULTS: Measurements/calculations of global hemodynamics (intravascular pressures, e.g., mean arterial pressure, heart rate, stroke volume index, cardiac output/index, left ventricular stroke work index, and systemic vascular resistance index), metabolism (oxygen delivery index and consumption index, oxygen extraction ratio, plasma lactate, hemoglobin), blood gases, pH, and hematocrit were made before fracture, immediately and 30 mins after fracture, and at 30-min intervals for the next 4 hrs. Hind-limb muscle water content was determined by dessication. Femur fracture led to acute reductions in cardiac output/index, stroke volume index, and oxygen delivery index and increases in systemic vascular resistance index and oxygen extraction ratio. In the absence of fluid resuscitation, these changes persisted and were accompanied by hypotension. Normal resuscitation attenuated the fracture-induced changes such that the only differences during resuscitation between limited resuscitation and normal resuscitation were a reduction in hematocrit in the latter and an increase in oxygen extraction ratio in the former. Water content increased in both injured and uninjured muscle. CONCLUSIONS: The cardiovascular and metabolic changes associated with femur fracture in the anesthetized pig can be reversed or prevented by crystalloids given in a volume equivalent to Advanced Trauma Life Support guidelines. PMID- 11889317 TI - Liquid ventilation with perflubron in the treatment of rats with pneumococcal pneumonia. AB - OBJECTIVE: To determine the efficacy of liquid ventilation using a medical-grade perfluorocarbon (perflubron) combined with parenteral or intratracheal antibiotics in a rat model of pneumonia. DESIGN: Prospective, laboratory investigation. SETTING: Experimental laboratory in a university medical center. SUBJECTS: Wistar rats (n = 112). INTERVENTIONS: One day after intratracheal inoculation with Streptococcus pneumoniae, rats received one of five experimental treatments or no treatment (control): modified liquid ventilation (MLV), intramuscular ampicillin, MLV plus intramuscular ampicillin, MLV with intratracheal ampicillin, or MLV plus ampicillin PulmoSpheres. MEASUREMENTS AND MAIN RESULTS: Animals receiving MLV plus intramuscular ampicillin, MLV with intratracheal ampicillin, or MLV plus ampicillin PulmoSpheres had significantly improved 10-day survival rates (85%, 72%, and 72%, respectively) compared with all other groups (0% to 25%). CONCLUSIONS: MLV in combination with either intramuscular, intratracheal, or PulmoSpheres ampicillin improved survival as compared with MLV alone or the same dose of antibiotics delivered intramuscularly. PMID- 11889319 TI - Elemental and intravenous total parenteral nutrition diet-induced gut barrier failure is intestinal site specific and can be prevented by feeding nonfermentable fiber. AB - OBJECTIVE: Parenteral nutrition and elemental diets both cause bacterial translocation, immune dysfunction, and increased infection in laboratory animals, whereas elemental diets, with or without fiber, ameliorate some, but not all gut barrier failure. The purpose of this study is to investigate, in an Ussing chamber system, whether elemental vs. parenteral diets induce gut barrier failure in specific anatomical sites in the intestine and whether fiber can ameliorate this phenomenon. DESIGN: Controlled study in laboratory animals. SETTING: University laboratory. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Nutritional support was provided to rats for 7 days by oral total parenteral nutrition (TPN; elemental diet) 307 kcal/kg/day, intravenous TPN (parenteral diet) 307 kcal/kg/day via jugular venous catheters, or rodent chow (controls). MEASUREMENTS AND MAIN RESULTS: Permeability to bacteria in intestinal segments of ileum, jejunum, and colon was evaluated in an Ussing chamber. Results were correlated with bacterial translocation to the mesenteric lymph nodes. Intravenous TPN caused greater bacterial translocation in all small intestinal segments and the cecum when compared with chow (p <.05). Oral TPN caused gut barrier failure only in the ileal segment, but not in the remainder of the small intestine (p <.001). Addition of cellulose provided a greater protection of the ileum to permeability than did pectin (p <.01). CONCLUSIONS: TPN causes global intestinal barrier failure, but elemental diet prevents barrier failure in parts of the small intestine other than the ileum. The addition of cellulose fiber to elemental diet can ameliorate further barrier failure in the ileum. PMID- 11889320 TI - Pulmonary capillary pressure during acute lung injury in dogs. AB - OBJECTIVES: To measure pulmonary capillary pressure and pulmonary artery occlusion pressures both during control conditions and during acute lung injury and to evaluate the effects of inotropic therapy and volume loading on these measurements after lung injury. DESIGN: Prospective, randomized, controlled laboratory trial. SETTING: University research laboratory. SUBJECTS: Eighteen heartworm-free mongrel dogs. INTERVENTIONS: Dogs were anesthetized (sodium pentobarbital, 30 mg/kg intravenously), intubated, and mechanically ventilated. A femoral artery and vein and the right external jugular vein were cannulated. After a median sternotomy, two pulmonary artery catheters were inserted via the jugular vein into the left and right lower lobar pulmonary arteries. Oleic acid (0.03 mL/kg) was administered to all dogs via the left pulmonary artery catheter, whereas the right lower lobe served as control. A baseline group of dogs received no further interventions, whereas two additional groups were given dobutamine (30 60 microg x kg(-1) x min(-1)intravenously) or saline boluses (1-2 L) before measurements were obtained after oleic acid lung injury. MEASUREMENTS AND MAIN RESULTS: Capillary pressure was estimated in both lower lung lobes by using the pulmonary artery occlusion method. Pulmonary capillary and pulmonary artery occlusion pressures were measured before and 2 hrs after oleic acid administration. Left lower lobar capillary pressure increased in all three groups, as did the difference between capillary pressure and pulmonary artery occlusion pressure. Capillary pressure in the control right lower lobe increased significantly only in the saline-loaded dogs, whereas the difference between the right-sided capillary and occlusion pressures increased only in the dogs given dobutamine. CONCLUSIONS: Oleic acid lung injury increases pulmonary capillary pressure independent of pulmonary artery occlusion pressure. The gradient between the two pressures was not significantly affected by volume loading or dobutamine infusion. PMID- 11889321 TI - Postischemic inotropic support of the dysfunctional heart. AB - OBJECTIVE: To determine relative adenine nucleotide regeneration and improvement in left ventricular (LV) function using three commonly used adrenergic agents- epinephrine, dobutamine, and phenylephrine---during reperfusion after a period of global ischemia. After initial resuscitation from cardiac arrest, adrenergic agents are frequently required to support postischemic LV dysfunction. However, the relative effectiveness and associated bioenergetic changes associated with these agents in the postischemic heart are unclear. DESIGN: Prospective, controlled laboratory study. SETTING: University research laboratory. SUBJECTS: Isolated, perfused Sprague-Dawley rat hearts. INTERVENTIONS: After 20 mins of global ischemia, isolated rat hearts were reperfused for 30 mins with Krebs Henseleit solution alone (control, n = 8), or with the addition of equipotent doses of epinephrine 1 microM (n = 8), dobutamine 0.3 microM (n = 8), or phenylephrine 50 microM (n = 8). In a second experiment, an alpha-1 antagonist, prazosin was given with phenylephrine to block the presumed alpha-1 agonist effect of phenylephrine. MEASUREMENTS AND MAIN RESULTS: A constant volume balloon was placed in the left ventricle to measure LV pressure and derived parameters of LV function. Adenine nucleotide concentrations were derived at various time points using high-performance liquid chromatography. During reperfusion, the phenylephrine group had significant improvement in LV function and cardiac efficiency in contrast to epinephrine and dobutamine. Total adenine nucleotides tended to be highest in the phenylephrine group with significant increases in adenosine diphosphate and adenosine monophosphate and no significant loss of adenosine triphosphate. The phenylephrine-induced increase in heart rate and developed pressure could be blocked with an alpha-1 antagonist, prazosin. CONCLUSIONS: In the isolated reperfused heart, phenylephrine, mediated by alpha-1 agonism, significantly improves postischemic LV dysfunction without worsening the overall myocardial metabolic state. PMID- 11889322 TI - Oxidative stress after reperfusion with primary coronary angioplasty: lack of effect of glucose-insulin-potassium infusion. AB - OBJECTIVE: To evaluate the oxidative stress status and the modification with glucose-insulin-potassium (GIK) therapy in patients with acute myocardial infarction undergoing primary percutaneous transluminal coronary angioplasty. DESIGN: Prospective, randomized, double-blinded, placebo-controlled study. SETTING: Cardiac intensive care unit at the university hospital. PATIENTS: Twenty patients were randomized to GIK solution (30% glucose in water with insulin 50 IU/L, and KCl 40 mM) vs. placebo (normal saline) at 1.5 mL/kg/hr for 24 hrs. The control group was 15 healthy volunteers with no heart disease. INTERVENTIONS: Eligible patients were randomized by a blinded pharmacist, patients with acute myocardial infarction were treated by primary percutaneous transluminal coronary angioplasty and randomized to GIK or placebo (saline solution). Primary angioplasty was successful in nine of ten patients (90%) and ten of ten patients (100%) in the GIK and placebo groups, respectively. Nine (100%) and six (60%) patients from GIK and placebo groups, respectively, underwent stent implantation. MEASUREMENTS AND MAIN RESULTS: We determined plasma levels of lipid peroxidation estimated by the malondialdehyde assay, superoxide dismutase, glutathione peroxidase, and catalase erythrocyte activities at admission and 0.5 and 24 hrs after angioplasty. Baseline determinations were compared with a control group (n = 15). Baseline clinical characteristics and time to treatment (4.5 +/- 3.5 hrs) were similar between groups. Angioplasty success rate (Thrombolysis in Myocardial Infarction [TIMI] 3 flow with residual stenosis < or = 30%) was 90% and 100% in GIK and placebo groups, respectively. Patients with acute myocardial infarction had an increase of malondialdehyde at baseline (2.9 +/- 1.7 vs. 1.1 +/- 0.3 microM, p <.01) and lower enzymatic activities of superoxide dismutase (0.5 +/- 0.5 vs. 1.3 +/- 0.4 U/mg hemoglobin, p <.01) and catalase (147 +/- 73 vs. 198 +/- 31 U/g hemoglobin, p <.01). These measurements did not change significantly after angioplasty and no differences were observed between GIK and placebo groups. CONCLUSION: Patients with acute myocardial infarction had increased levels of oxidative stress associated with a reduction in enzymatic antioxidant reserve. Administration of GIK solution did not improve these abnormalities among patients undergoing primary angioplasty. PMID- 11889324 TI - The ability of endotoxin-stimulated enterocytes to produce bactericidal factors. AB - OBJECTIVE: Bactericidal peptides, specifically defensins, are produced by polymorphonuclear cells. Intestinal epithelial cells also produce bactericidal peptides, perhaps as part of their barrier function, to the greatest load of endogenous bacteria present in the body. We sought to determine whether and under what conditions intestinal cell lines could produce bactericidal compounds. DESIGN: Laboratory investigation. SETTING: Children's burn hospital. SUBJECTS: Caco-2, IEC-6, and HT-29 cell lines. INTERVENTIONS: Three different enterocyte lines were cultured for 1 day +/- lipopolysaccharide (1 or 10 microg/mL), and their supernatants were tested for bactericidal activity. Also, reverse transcription-polymerase chain reaction of Caco-2 cells was performed to assess the expression of defensin-6 mRNA. MEASUREMENTS AND MAIN RESULTS: After culture, enterocytes all were found to release one or more soluble factors with bactericidal activity (as measured fluorometrically by using a metabolizable dye) when stimulated by lipopolysaccharide (1 microg/mL). The bactericidal activity of these culture supernatants was saturated by increased bacterial load, additive to the effects of normal human peripheral blood polymorphonuclear cells, and was reduced by serial supernatant dilution. Enterocyte stimulation with larger amounts of lipopolysaccharide (10 microg/mL) resulted in greater bactericidal activity. After supernatant fractionation based on molecular weight, the bactericidal effect was best retained in the <10-kDa fraction. In addition, the expression of mRNA for defensin-6, a bactericidal peptide produced by neutrophils, was seen in Caco-2 cells. CONCLUSION: Enterocytes are shown to produce a soluble, low molecular weight, bactericidal compound in response to endotoxin stimulation. The expression of defensin-6 mRNA in Caco-2 cells suggests that intestinal cells may release defensins as bactericidal peptides. This experimental system provides an in vitro model to study the activity and production of bactericidal factors by enterocytes. PMID- 11889323 TI - Effects of perfluorohexane vapor on relative blood flow distribution in an animal model of surfactant-depleted lung injury. AB - OBJECTIVE: To test the hypothesis that treatment with vaporized perfluorocarbon affects the relative pulmonary blood flow distribution in an animal model of surfactant-depleted acute lung injury. DESIGN: Prospective, randomized, controlled trial. SETTING: A university research laboratory. SUBJECTS: Fourteen New Zealand White rabbits (weighing 3.0-4.5 kg). INTERVENTIONS: The animals were ventilated with an FIO(2) of 1.0 before induction of acute lung injury. Acute lung injury was induced by repeated saline lung lavages. Eight rabbits were randomized to 60 mins of treatment with an inspiratory perfluorohexane vapor concentration of 0.2 in oxygen. To compensate for the reduced FIO(2) during perfluorohexane treatment, FIO(2) was reduced to 0.8 in control animals. Change in relative pulmonary blood flow distribution was assessed by using fluorescent labeled microspheres. MEASUREMENTS AND MAIN RESULTS: Microsphere data showed a redistribution of relative pulmonary blood flow attributable to depletion of surfactant. Relative pulmonary blood flow shifted from areas that were initially high-flow to areas that were initially low-flow. During the study period, relative pulmonary blood flow of high-flow areas decreased further in the control group, whereas it increased in the treatment group. This difference was statistically significant between the groups (p =.02) as well as in the treatment group compared with the initial injury (p =.03). Shunt increased in both groups over time (control group, 30% +/- 10% to 63% +/- 20%; treatment group, 37% +/- 20% to 49% +/- 23%), but the changes compared with injury were significantly less in the treatment group (p =.03). CONCLUSION: Short treatment with perfluorohexane vapor partially reversed the shift of relative pulmonary blood flow from high flow to low-flow areas attributable to surfactant depletion. PMID- 11889326 TI - Regional intraparenchymal pressure differences in experimental intracerebral hemorrhage: effect of hypertonic saline. AB - OBJECTIVE: To study regional intraparenchymal pressures within the cranial cavity during and after formation of intracerebral hemorrhage. We also assessed the effect of hypertonic saline on intraparenchymal pressure in different brain regions and on regional brain distribution of sodium within the brain. DESIGN: Prospective, controlled, laboratory trial. SETTINGS: Animal research laboratory. SUBJECTS: Eight mongrel dogs, weighing 15-25 kg. INTERVENTION: We introduced an intracerebral hematoma in eight mongrel dogs by infusing 6 mL of autologous arterial blood in the deep white matter adjacent to the basal ganglia. Sodium chloride (23.4%, 1.4 mL/kg) then was administered intravenously 6 hrs after introduction of hematoma. MEASUREMENTS AND MAIN RESULTS: Parenchymal pressure monitors were placed in the perihematoma region, both frontal lobes, and the cerebellum to record intraparenchymal pressure during and 6 hrs after intracerebral hematoma formation. Intraparenchymal pressure measurements were recorded for 3 more hours after administration of 23.4% sodium chloride. Regional cerebral perfusion pressure was calculated for each intraparenchymal pressure measurement. Regional sodium distribution was measured in extracts from brain regions by using ion selective electrode technique. A higher elevation in intraparenchymal pressure was recorded in the perihematoma region during the introduction of the hematoma compared with other compartments. After 5 mL of autologous blood was introduced, intraparenchymal pressure (mm Hg +/- SE) was significantly higher in the perihematoma region (42.1 +/- 3.5) than in the ipsilateral (30.0 +/- 4.6, p <.05) and contralateral (27.1 +/- 5.5, p <.01) frontal lobes and cerebellum (29.1 +/- 4.5, p <.05). Four hours after introduction of the hematoma, the cerebral perfusion pressure recorded in the perihematoma region (43.6 +/- 9.7) remained significantly lower than in the ipsilateral (58.6 +/- 9.3, p <.05) but not the contralateral frontal lobes (54.7 +/- 10.1) and cerebellum (51.0 +/- 11.1). Administration of 23.4% sodium chloride immediately reduced intraparenchymal pressure in each compartment. This effect was still observed at 3 hrs in each compartment. Sodium concentration was higher in the perihematoma region than in the frontal lobes, cerebellum, or brain stem. CONCLUSIONS: Prominent differences were observed in intraparenchymal pressure and cerebral perfusion pressure in the perihematoma region and frontal lobes during and after intracerebral hematoma. We speculate that the potential importance of regional intraparenchymal pressure differences in the clinical settings may be under appreciated. In this canine model of intracerebral hematoma, a single dose of hypertonic saline effectively reduces the intraparenchymal pressure in all regions of the brain. PMID- 11889327 TI - Beneficial effects of nitric oxide inhalation on pulmonary bacterial clearance. AB - OBJECTIVE: The beneficial effects of nitric oxide inhalation on oxygenation during acute respiratory distress syndrome are well described. In contrast, the effects of nitric oxide on pulmonary inflammatory response are much less known in vivo. The objectives of this study were to evaluate the effects of nitric oxide inhalation on bacterial clearance during bacterial pneumonia and on alveolar neutrophil functions. DESIGN: Controlled animal study. SETTING: Research laboratory of an academic institution. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Severe pneumonia was induced by alveolar instillation of live Pseudomonas aeruginosa (1.5 x 10(8) colony-forming units/kg) in rats. After instillation, rats were exposed to oxygen alone (FIO(2) 100%) or to oxygen (FIO(2) approximately 100%) plus nitric oxide (10 ppm) during 24 hrs. MEASUREMENTS AND MAIN RESULTS: Oxygen plus nitric oxide inhalation compared with oxygen alone increased recruitment of alveolar neutrophils (32.5 +/- 4.6 x 10(6) cells/mL vs. 23.4 +/- 1.9 x 10(6) cells/mL, p <.05) and improved bacterial clearance in the bronchoalveolar lavage fluid (8.1 +/- 4.2 x 10(2) vs. 1.6 +/- 1.0 x 10(5) colony-forming units/mL, p <.05) and in the pulmonary parenchyma (1.7 +/- 1.14 x 10(7) vs. 3.4 +/- 1.5 x 10(8) colony-forming units/mL, p <.05). However, neither protein concentration in the bronchoalveolar lavage fluid nor mortality rates were modified by nitric oxide inhalation. The ex vivo alveolar neutrophil functions were similar regardless of whether rats previously inhaled nitric oxide. In vitro experiments demonstrated that nitric oxide donor had a direct bactericidal effect against P. aeruginosa and did not modify alveolar neutrophil functions. CONCLUSIONS: These results suggest a beneficial effect of nitric oxide inhalation on bacterial clearance of P. aeruginosa attributable to both a direct bactericidal effect and an influx of alveolar neutrophils with preserved functions. PMID- 11889328 TI - Randomized, double-blind, placebo-controlled trial of intravenous salbutamol and nebulized ipratropium bromide in early management of severe acute asthma in children presenting to an emergency department. AB - BACKGROUND: In acute severe asthma, treatment must be initiated early to reverse the pathophysiology that may render airways less responsive to bronchodilation. The addition of nebulized ipratropium bromide to initial emergency department therapy improves pulmonary function, but it is unclear whether this approach results in earlier hospital discharge. The early use of bolus intravenous salbutamol has also been shown to improve outcome, including earlier discharge. We therefore assessed the relative benefits of intravenous salbutamol and nebulized ipratropium bromide in the early management of acute severe asthma in children by a double-blind, randomized, controlled trial. METHODS: This study was undertaken at a tertiary children's hospital, The Children's Hospital at Westmead, The Royal Alexandra Hospital for Children, Westmead, Sydney, Australia. Only children with severe acute asthma as determined by the National Asthma Campaign guidelines criteria and pulmonary index were included. All children received initial nebulized salbutamol therapy (2.5-5 mg salbutamol in 4 mL of normal saline depending on age) at initial emergency department presentation. If asthma remained severe 20 mins later, an intravenous cannula was inserted and intravenous methylprednisolone (1 mg/kg) was administered to all children receiving nebulized salbutamol every 20 mins. Children were then randomized to one of three groups: intravenous salbutamol (15 microg/kg as a single bolus over 10 mins), ipratropium bromide (250 microg), or intravenous salbutamol plus ipratropium bromide. All observers were blinded to treatment groups. Children were randomly assigned to receive a single-dose intravenous bolus of either saline or salbutamol and either nebulized saline or ipratropium bromide determined by a number generated randomly in the hospital pharmacy. The primary outcomes were recovery time and discharge time of each group. Respiratory and hemodynamic monitoring were continuous during the first 2 hrs of the study and then children were monitored clinically for 24 hrs. RESULTS: A total of 55 children with acute severe asthma were entered into the study over an 18-month period. The three groups were similar demographically, with a mean age of 5.9 yrs, and mean duration of attack of 19.6 hrs. No side effects or treatment intolerance were reported. Children in the groups that received intravenous salbutamol had a significant reduction in recovery time to achieving second hourly inhaled salbutamol (p =.008) compared with those administered inhaled bronchodilator alone. The addition of ipratropium bromide to intravenous salbutamol provided no significant further benefit in terms of nebulizer therapy (intravenous salbutamol compared with intravenous salbutamol plus ipratropium bromide). Children administered intravenous salbutamol ceased supplemental oxygen therapy earlier than those administered ipratropium alone at 12 hrs post randomization (p =.0003). Children administered intravenous salbutamol could be discharged from the hospital 28 hrs earlier than those administered ipratropium bromide (p =.013). CONCLUSION: Children administered intravenous salbutamol for severe acute asthma showed a more rapid recovery time, which resulted in earlier discharge from the hospital than those administered inhaled ipratropium bromide. There was no additional benefit obtained by combining ipratropium bromide and intravenous salbutamol administration. PMID- 11889329 TI - Complications of central venous catheters: internal jugular versus subclavian access--a systematic review. AB - OBJECTIVE: To test whether complications happen more often with the internal jugular or the subclavian central venous approach. DATA SOURCE: Systematic search (MEDLINE, Cochrane Library, EMBASE, bibliographies) up to June 30, 2000, with no language restriction. STUDY SELECTION: Reports on prospective comparisons of internal jugular vs. subclavian catheter insertion, with dichotomous data on complications. DATA EXTRACTION: No valid randomized trials were found. Seventeen prospective comparative trials with data on 2,085 jugular and 2,428 subclavian catheters were analyzed. Meta-analyses were performed with relative risk (RR) and 95% confidence interval (CI), using fixed and random effects models. DATA SYNTHESIS: In six trials (2,010 catheters), there were significantly more arterial punctures with jugular catheters compared with subclavian (3.0% vs. 0.5%, RR 4.70 [95% CI, 2.05-10.77]). In six trials (1,299 catheters), there were significantly less malpositions with the jugular access (5.3% vs. 9.3%, RR 0.66 [0.44-0.99]). In three trials (707 catheters), the incidence of bloodstream infection was 8.6% with the jugular access and 4.0% with the subclavian access (RR 2.24 [0.62-8.09]). In ten trials (3,420 catheters), the incidence of hemato- or pneumothorax was 1.3% vs. 1.5% (RR 0.76 [0.43--1.33]). In four trials (899), the incidence of vessel occlusion was 0% vs. 1.2% (RR 0.29 [0.07-1.33]). CONCLUSIONS: There are more arterial punctures but less catheter malpositions with the internal jugular compared with the subclavian access. There is no evidence of any difference in the incidence of hemato- or pneumothorax and vessel occlusion. Data on bloodstream infection are scarce. These data are from nonrandomized studies; selection bias cannot be ruled out. In terms of risk, the data most likely represent a best case scenario. For rational decision-making, randomized trials are needed. PMID- 11889331 TI - Randomized, controlled clinical trials in sepsis: has methodological quality improved over time? AB - OBJECTIVE: To systematically evaluate the methodological quality of randomized clinical trials and to determine whether randomized clinical trials of sepsis improved in methodological quality over time. DATA SOURCES: Computerized MEDLINE search of articles published in any language from 1966 to 1998 combined with a manual search of bibliographies of published articles and communication with known experts in the field. STUDY SELECTION: All randomized clinical trials of sepsis, severe sepsis, and septic shock performed in adults and published as full articles. DATA EXTRACTION: Abstracts of all retrieved records were reviewed and the inclusion criteria were applied. All selected articles were classified into (a) trials designed to detect differences in mortality as the primary end point, or (b) trials focusing on surrogate outcome measures (i.e., physiological or biochemical parameters). All retrieved trials were then graded for methodological quality using an objective grading scheme developed specifically for this study. The data selection and extraction process was carried out independently by two of the authors; any disagreement was resolved by discussion. DATA SYNTHESIS: Seventy four randomized clinical trials involving septic patients qualified for inclusion in this study (40 reporting mortality outcomes, 34 reporting other surrogate outcomes). Trials reporting mortality as the primary outcome had significantly higher quality scores compared with trials reporting surrogate outcome measures (29.6 +/- 1.0 vs. 24.3 +/- 0.8, p =.0006). From 1976 to 1998, trial methodology improved significantly over time (an average of 0.36 points per year, p =.021). Mortality outcome trials improved an average of 0.58 points per year (p =.0011) whereas surrogate outcome trials did not demonstrate an improvement in methodological quality over time (p =.249). CONCLUSION: The methodological limitations identified in this article can help to target further improvement in trial design to enhance the validity of findings from future randomized clinical trials of sepsis. PMID- 11889332 TI - Serotonin syndrome presenting as hypotonic coma and apnea: potentially fatal complications of selective serotonin receptor inhibitor therapy. AB - OBJECTIVE: To describe a patient who developed serotonin syndrome on four separate occasions as a result of monotherapy with two different selective serotonin receptor inhibitors (fluoxetine and cetalopram). DESIGN: Case report. SETTING: Community hospital. PATIENTS: Single patient with four episodes of serotonin syndrome. MEASUREMENTS AND MAIN RESULTS: The syndrome was characterized by coma/unresponsiveness (four episodes), dilated pupils (four episodes), salivation (two episodes), dryness of mouth (two episodes), myoclonus like activity of eyelids (four episodes), oculogyric crisis (four episodes), flaccid paralysis of all extremities (four episodes), tremors (two episodes), apnea (two episodes), restlessness (one episode). Recovery occurred within 24 hrs, although muscle pain and weakness persisted for 2 months after stopping fluoxetine. Apnea occurred in both episodes associated with fluoxetine therapy. CONCLUSION: Apnea and coma may occur in serotonin syndrome. PMID- 11889333 TI - Severe status asthmaticus: management with permissive hypercapnia and inhalation anesthesia. AB - OBJECTIVE: To describe the difficulties that can be encountered during mechanical ventilation of severe status asthmaticus and to discuss the safety of permissive hypercapnia as a ventilatory strategy and the role and limitations of inhalation anesthesia in the treatment of refractory cases. DESIGN: Case series and review of literature. SETTING: Intensive care unit of a tertiary care hospital. PATIENTS: Two patients with severe status asthmaticus. INTERVENTIONS: Administration of inhalational anesthetics. MEASUREMENTS AND MAIN RESULTS: Both patients had respiratory failure secondary to status asthmaticus requiring mechanical ventilation and permissive hypercapnia. They also received inhalational anesthetics because of refractory bronchoconstriction. Levels of PaCO(2) in each case were among the highest and most prolonged elevations (>150 mm Hg for several hours) reported to date. In one case, life-threatening difficulties with ventilation were encountered related to the use of an anesthesia ventilator. Although they had complications related to the severity of their illnesses, both were treated to recovery. CONCLUSIONS: Mechanical ventilation in severe status asthmaticus can be challenging. Permissive hypercapnia is a relatively safe strategy in the ventilatory management of asthma. High levels of hypercapnia and associated severe acidosis are well tolerated in the absence of contraindications (i.e., preexisting intracranial hypertension). Inhalation anesthesia may be useful in the treatment of refractory cases of asthma but should be used carefully because it may be hazardous owing to poor flow capabilities of most anesthesia ventilators. PMID- 11889334 TI - A predilection for posterior prediction and phenotypic precision. PMID- 11889336 TI - Gastrointestinal tissue capnometry and critical oxygen delivery: flow versus hypoxia. PMID- 11889335 TI - Tissue dysoxia in sepsis: getting to know the mitochondrion. PMID- 11889337 TI - The importance of alpha-adrenergic stimulation to the postischemic heart. PMID- 11889338 TI - Jugular versus subclavian central venous catheter insertion: search for the better approach. PMID- 11889339 TI - Assessing methodological quality of clinical trials in sepsis: searching for the right tool. PMID- 11889340 TI - Ethics consultations in the intensive care setting. PMID- 11889341 TI - Route of feeding in critically ill patients. PMID- 11889342 TI - The central nervous system hypothalamic-pituitary-adrenal axis in sepsis. PMID- 11889343 TI - Hyperprocalcitonemia in patients with perioperative myocardial infarction after cardiac surgery. PMID- 11889345 TI - Colicky infants according to maternal reports in telephone interviews and diaries: a large Scandinavian study. AB - In this population-based study the colic incidence was 9.4%, according to telephone interviews with the parents made when the infants were 5 weeks of age (n = 1628), and parental concern about infant crying was common. However, 7-day diaries of colicky and control infants (n = 116 + 119) revealed low distress amounts in colicky infants in general. In the subgroup of Wessel-colicky infants (n = 37), distress episodes were frequent and long lasting, and there was a high proportion of colicky crying versus fussing and normal crying. Even if there may be a reduction in the infantile colic incidence and support for the hypothesis that infantile colic is at least partially "in the eye of the beholder," that is, the concerned parent, a subgroup of infants may be more "genuinely colicky." Women who had stated in late-pregnancy interviews that there is a risk of spoiling an infant with too much physical contact were more likely to have infants with colic, and their infants were more distressed, even when given the same amount of physical contact. This finding warrants further elucidation. PMID- 11889346 TI - Developmental coordination disorder in extremely low birth weight children at nine years. AB - Developmental coordination disorder (DCD) is defined as an impairment in the development of motor coordination that interferes with academic achievement or activities of daily living (DSM-IV). DCD has been reported to affect 5% to 9% of children in the normal population. This study describes the prevalence of DCD in a cohort of extremely low birth weight children (ELBW, < or = l800 g) at 8.9 years of age, from which were excluded children with major impairments. Seventy three children were included in the study group, along with 18 term-born, socially matched controls. Of the 73 ELBW children, 37 (51%) were classified as having DCD. ELBW children with DCD also had significantly lower Performance IQ (PIQ) scores and were more likely (43%) to have a learning difficulty in arithmetic than ELBW children who did not have DCD. This study found that DCD is a common problem in school-aged ELBW children. PMID- 11889347 TI - Impact of low birth weight on attention-deficit hyperactivity disorder. AB - The objective of the study was to evaluate an association between low birth weight (LBW) and attention-deficit hyperactivity disorder (ADHD) attending to potential family-genetic and environmental confounders. We examined 252 ADHD cases (boys and girls) and 231 non-ADHD controls and their parents. All subjects were extensively assessed with structured diagnostic interviews, cognitive assessments, and structured interviews of prenatal, infancy, and delivery complications. ADHD cases were three times more likely to have been born LBW than were non-ADHD controls, after attending to potential confounders such as prenatal exposure to alcohol and cigarettes, parental ADHD, social class, and comorbid disruptive behavior disorders in parents and offspring. If this association was causal, 13.8% of all ADHD cases could be attributed to LBW. These results converge with prior studies documenting similar associations and indicate that LBW is an independent risk factor for ADHD. Children with LBW, however, make up a relatively small proportion of children with ADHD. PMID- 11889349 TI - Cluster analysis of maternal characteristics and perceptions of child behavior problems in a behavioral pediatrics practice. AB - Mothers bringing their children to a behavioral pediatrics clinic vary considerably in terms of concerns about their children, their own emotional status, and their sense of familial and social support. Knowledge of these factors may enhance differential diagnosis and advise treatment decisions. Mothers of 90 children ages 6-12 years completed the Child Behavior Checklist (CBCL), Mental Health Inventory (MHI), Dyadic Adjustment Scale (DAS), and Health Concerns Questionnaire before their initial appointment. Cluster analysis revealed four groups of mothers that varied in their apparent motivation for seeking assistance. These groups included advice-seeking mothers, mothers that had concerns about the medical well-being of their children, mothers that were overwhelmed by their current circumstances, and mothers whose concerns about their dyadic relationships may have been displaced onto their children. The study findings support the use of cluster analysis in clinical research. Future research could focus on the specific intervention needs of these different types of families. PMID- 11889350 TI - Fatigue, decreased interest in play, motor delay, and elevated liver function tests in a 4-year-old boy. PMID- 11889348 TI - Patterns of psychotropic medication use in very young children with attention deficit hyperactivity disorder. AB - Psychotropic medications are increasingly used for very young children. Patterns of use in a well-described group of children 3 years and younger with a diagnostic label of attention-deficit hyperactivity disorder (ADHD) reveal both reasons to use such medications and concerns about how these medications are used. Of 223 children with ADHD, more than half (n = 127) received psychotropic medications in an idiosyncratic manner, both in the specific medication and in use over time. Almost half of the children who were medicated did not have opportunities for monitoring as often as every 3 months, despite the fact that more than half received psychotropic medications for 6 months or longer. Children with comorbid mental health conditions and chronic health conditions were at greater risk for receiving psychotropic medications. These patterns of use demonstrate a compelling need for guidance in psychopharmacological treatment of very young children. PMID- 11889351 TI - Pediatric bipolar mood disorder. AB - The diagnosis of bipolar mood disorder (BP) in preadolescents (pediatric mania) has generated considerable controversy in terms of its estimated prevalence and validity as a diagnostic category. The relative paucity of systematic studies and the current diagnostic confusion related to the disorder are often attributed to the apparent discontinuities in the childhood versus adult presentation of the illness, namely, irritability as the predominant "mood" of mania and a continuous course of symptoms. The goal of this article is to review the current literature and identify sources of confusion relating to pediatric mania by considering results to date within a larger context that include findings from studies on (1) BP illness in adults, (2) mood disorders across the lifespan, (3) the role of development in symptom expression, and (4) patterns of heritability in psychiatric disorders. Whereas much remains to be investigated in the validation of the diagnosis for children, integrating results across studies may provide a framework for understanding the differences in the presentation of severe mood disorders in children and adults. PMID- 11889358 TI - The structure and invariance of a model of social functioning in schizophrenia. AB - The authors proposed and tested a model of social functioning in schizophrenia. The model consisted of five indicators: social competence, quality of relations, satisfaction, symmetry of relationships, and the number of close friends. The model combines proximal and distal, quality and quantity, and self-report and observer ratings of social functioning. It was designed to have ecological validity in that it reflects real-world social functioning, and the data are all gathered in reference to naturally occurring social contexts. Two independent community samples of individuals diagnosed with schizophrenia or schizoaffective disorder (N = 172; N = 166) were used to test the structure of the measurement model and its invariance across samples. Additional measures of social functioning, prognosis, and intrapsychic deficits were used to further assess the validity of the proposed social functioning construct. A single-factor model had acceptable fit with the data from sample 1. The single-factor model was found to be invariant across the two independent samples. Correlations with other measures in both samples provided additional evidence for the construct validity of the proposed measurement model of social functioning. Implications for research and practice are discussed. PMID- 11889359 TI - Object relations deficits in schizophrenia: a cross-cultural comparison between Brazil and the United States. AB - Object relations deficits are commonly found in schizophrenia samples from the United States, but it is unknown whether these deficits are a reliable finding in other cultures. The Bell Object Relations Inventory was translated into Brazilian Portuguese and administered to 61 stable outpatients with schizophrenia from Sao Paolo, Brazil. Their scores were compared with a Brazilian normal sample and with a matched U.S. schizophrenia sample. The Brazilian normal sample showed a pattern of scores within the normal range when compared with U.S. norms. The Brazilian schizophrenia sample had significantly greater pathology than the Brazilian normal sample on Alienation, Egocentricity, and Social Incompetence. Their mean scores on Alienation were similar to the matched U.S. schizophrenia sample, and they had significantly greater pathology on Insecure Attachment, Egocentricity, and Social Incompetence with 85.6% showing some type of object relations deficit. Findings support the cross-cultural validity of the Bell Object Relations Inventory and the ubiquity of object relations deficits in schizophrenia. The most common profiles for both schizophrenia samples were the Psychotically Egocentric and the Socially Withdrawn object relations types. PMID- 11889360 TI - Subjective experiences of Japanese inpatients with chronic schizophrenia. AB - Knowledge of the subjective experience of patients has important implications for the understanding and treatment of schizophrenia. However, previous studies examining the clinical correlates of subjective experiences have yielded inconclusive results. The present study of Japanese inpatients with chronic schizophrenia aimed to reveal the factorial structure of patients' subjective experiences and to examine the relationships between identified factors and demographic and clinical variables. A systematic assessment was carried out of objective psychopathology and subjective experiences in 129 inpatients. A factor analysis of the data of patients' subjective experiences identified five factors. Delusions/hallucinations, as clinical variables, were significantly associated with three of these factors. Negative symptoms, extrapyramidal side effects, and female gender each had significant associations with only one of the factors. The first factor, which accounted for the highest percentage, did not correlate significantly with any of the clinical or demographic variables. Subjective experiences in chronic schizophrenia appear to have a heterogeneous structure and fall into two groups, one relatively independent of objective psychopathology and the other reflecting the clinical variable delusions/hallucinations. PMID- 11889361 TI - Adult religiousness and history of childhood depression: eleven-year follow-up study. AB - This study investigates the association between childhood depression and the protective qualities of adult religiousness. Subjects were 146 (65 female and 81 male) adults with a history of childhood depression and 123 (61 female and 62 male) adults without a history of childhood depression interviewed as part of a long-term follow-up study (mean years of follow-up, 11.2; SD = 1.4). Depression in childhood and adulthood was assessed by blind and independent clinical interviews by using the Schedule for Affective Disorders for School Aged Children and the Schedule for Affective Disorders Life-time Version, respectively. Religiousness was assessed by report on the personal importance of religion, frequency of attendance of religious services, religious denomination, and child adult concordance of report. Findings showed adult personal importance of religion to be associated with a decreased risk for depression in women without a history of childhood depression but an increased risk for depression in women with a history of childhood depression. Adult Catholicism as compared with Protestantism was associated with a decreased risk for depression in male childhood depressives, but this association was not found in men without childhood depression. The findings potentially suggest a reciprocal-influence process between childhood pathology and the development of religiousness. PMID- 11889362 TI - Use of the Chinese version of the Beck Depression Inventory for screening depression in primary care. AB - Many Asian-Americans are unfamiliar with depression and its treatment. When depressed, they generally seek treatment from their primary care physicians and complain about their physical symptoms, resulting in under-recognition and under treatment of depression. This study evaluates the effectiveness of the Chinese version of the Beck Depression Inventory (CBDI) for screening depression among Chinese-Americans in primary care. A total of 503 Chinese-Americans in the primary care clinic of a community health center were administered the CBDI for depression screening. Patients who screened positive (CBDI > or = 16) were interviewed by a psychiatrist using the Structured Clinical Interview for DSM-III R, patient version (SCID-I/P) for confirmation of the diagnosis. Patients who screened negative (CBDI < 16) were randomly selected to be interviewed using the depression module of the SCID-I/P. The results of the SCID-I/P interview were used as the standard for evaluating the sensitivity and specificity of the CBDI. A total of 815 Chinese-Americans in a primary care clinic were approached, and 503 completed the CBDI. Seventy-six (15%) screened positive (CBDI > or = 16), and the prevalence of major depression was 19.6% by using extrapolated results from SCID-I/P interviews. When administered by a native-speaking research assistant, the CBDI has good sensitivity (.79), specificity (.91), positive predictive value (.79), and negative predictive value (.91). Despite the commonly believed tendency to focus on physical symptoms rather than depressed mood, Chinese Americans are able to report symptoms of depression in response to a questionnaire. The CBDI, when administered by research assistants, has good sensitivity and specificity in recognizing major depression in this population. Lack of interest among Chinese-American patients in using the CBDI as a self rating instrument has limited its use for depression screening in primary care settings. PMID- 11889363 TI - Psychosocial aspects of substance abuse by clients with severe mental illness. AB - As the literature on co-occurring substance abuse in persons with severe mental illnesses has evolved, emphasis on biologic and pharmacologic factors has diverted attention from important psychosocial issues. The authors review recent research showing that a) psychosocial risk factors may explain consistently high rates of substance abuse by these persons, b) substance abuse is for most clients a socio-environmental phenomenon embedded in interpersonal activities, and c) both natural recovery processes and effective treatments rely on developing new relationships, activities, coping strategies, and identities. Thus, psychosocial issues are critical in our attempts to understand and address substance abuse in this population. PMID- 11889364 TI - Psychiatric symptoms of opium and cocaine use. 1913. PMID- 11889365 TI - The experience of providing care to relatives with chronic mental illness. AB - Research on family caregivers of mentally ill relatives has historically focused on negative aspects of caregiving, often described as caregiver burden. The authors document caregivers' perspectives on both the negative and positive aspects of caregiving. A qualitative approach was used. Data collection involved 20 in-depth, audiotaped, semistructured interviews focusing on caregivers' positive and negative personal experiences with caregiving to a relative with mental illness. Caregivers reported common negative impacts but also beneficial effects, such as feelings of gratification, love, and pride. Main themes included stigma, systems issues, life lessons learned, and love and caring for the ill relative. This study counterbalances the predominantly negative consequences previously reported and adds to the emerging literature on positive aspects of caregiving. Mental health professionals need to help caregiving families make choices to improve their challenging situations and identify the rewards of caregiving, and to advocate for increased systemic supports to ease caregiver burden. PMID- 11889366 TI - Sociodemographic factors associated with attempted suicide in two Israeli cities between 1990 and 1998. PMID- 11889367 TI - Posttraumatic stress disorder after terrorist attacks: a review. PMID- 11889368 TI - Cultural mistrust predicts age at first hospitalization for African-American psychiatric patients. PMID- 11889370 TI - Full disclosure of financial interests in biomedical publications--a reminder. PMID- 11889371 TI - Otolaryngology retirement profile in the southeastern United States. AB - OBJECTIVE: To document retirement-related issues and trends among otolaryngologists. STUDY DESIGN: Survey of 438 retired members in the Southern geographical region of the American Laryngological, Rhinological and Otological Society, Inc. METHODS: A questionnaire was mailed to retired members, completed anonymously, and returned to the author. RESULTS: A total of 138 (31.5%) surveys were received. Respondents' average age at retirement was 63.2 years; approximately half had retired in the last 5 years. Since 1995, most had left either a group practice (45%) or a solo practice (41%). The majority of respondents (40%) retired for previously planned or personal reasons. Two thirds of respondents reported that they were more satisfied with retirement than expected. This greater satisfaction was seen in those with a higher average income after retirement. The most common advice for colleagues still practicing was to save more money and invest more money. CONCLUSIONS: Although these results are biased because of a self-selected group of respondents, they illustrate that for this group of retired otolaryngologists, although retirement is being planned for, it is not occurring earlier than in previous years. The experience of retirement was largely positive for these respondents. PMID- 11889372 TI - Telemedicine: teleproctored endoscopic sinus surgery. AB - OBJECTIVE/HYPOTHESIS: Teleproctored surgery projects a surgeon's expertise to remote locations. The objective of the present study was to evaluate the safety and feasibility of this technique as compared with the current standard of care. STUDY DESIGN: Prospective. METHODS: A study was conducted in a residency training program comparing conventionally proctored endoscopic sinus surgery cases with teleproctored cases, with the faculty surgeon supervising through audiovisual teleconferencing (VTC) in a control room 15 seconds from the operating room. RESULTS: Forty-two control patients (83 sides) and 45 teleproctored patients (83 sides) were evaluated. There were no internal differences between groups regarding extent of polypoid disease, revision status, procedures per case, degree of difficulty, general or local anesthesia, or microdebrider use. There were no cases of visual disturbance, orbital ecchymosis or hematoma, or cerebrospinal fluid leak. Orbital fat herniation and blood loss were equal between groups. Three teleproctored cases required faculty intervention: two for surgical difficulty, one for VTC problems. Teleproctored cases took 3.87 minutes longer per side (28.54 vs. 24.67 min, P <.024), a 16% increase. This was thought to be a result of nuances of VTC proctoring. Residents had a positive learning experience, with nearly full control of the operating suite combined with remote supervision through telepresence. Faculty thought such supervision was safe but had concerns regarding personal skills maintenance. CONCLUSIONS: Teleproctored endoscopic sinus surgery can be safely performed on selected cases with an acceptable increase in time. Teleproctored surgery with remote sites may continue to be safely investigated. Incorporating remote supervision through telepresence into the curriculum of surgical residency training requires further study. PMID- 11889373 TI - Risk factors and demographics in patients with spasmodic dysphonia. AB - OBJECTIVES: Spasmodic dysphonia has been characterized as a functional, psychogenic, or movement disorder with no known etiology or cure. In the present study, risk factors associated with other movement disorders were evaluated in patients with spasmodic dysphonia. STUDY DESIGN: Retrospective patient survey of 168 patients with a known diagnosis of spasmodic dysphonia who completed questionnaires at the time of interval botulinum toxin injection. METHODS: Patients completed questionnaires on demographics, education level, work history, significant life events, medical, social, and family history. The results were compared with those of first-degree relatives as a control group with similar demographics. Data were analyzed using percentages calculated on the total number of responses and distribution of frequency of each. Statistical significance was estimated on t tests of chi2 values. RESULTS: In the series of 168 patients, there was a female predominance of 79%. Age range at onset was 13 to 71 years with an average of age of 45 years. Sixty-five percent of patients had previously had the measles or mumps compared with the national average of 15% in a similar age group (P =.0001). Thirty percent of patients directly associated onset of spasmodic dysphonia symptoms to an upper respiratory tract infection, and 21% to a major life stress. There was no significant incidence of any other medical or neurological condition or symptomatology. There was no family history of spasmodic dysphonia. Twenty-six percent of patients had an essential tremor compared with 4% of first-degree relatives (P =.0001), and 11% had associated writer's cramp compared with 2% of relatives (P =.02). Less than 1% of patients described a history of toxic exposure or electrical injury. CONCLUSIONS: The majority of patients with spasmodic dysphonia are girls and women. A significantly higher incidence of childhood viral illness was found in the patients with spasmodic dysphonia. Patients with spasmodic dysphonia had a significant incidence of both essential tremor and writer's cramp but no history of major illness or other neurological disorder. There appear to be no significant environmental or hereditary patterns in the etiology of spasmodic dysphonia. Stress or viral infection may induce the onset of symptoms of spasmodic dysphonia. Many features of the disorder are common to other movement disorders, and this knowledge may direct future research efforts. PMID- 11889374 TI - The role of computed tomography and magnetic resonance imaging in patients with sinusitis with complications. AB - OBJECTIVES: This study compares the use of computed tomography (CT) with magnetic resonance imaging (MRI) and clinical assessment in the diagnosis and surgical management of patients with sinusitis with complications. The common CT findings of patients with intracranial or orbital infection of sinusitis are reviewed. STUDY DESIGN: We reviewed the charts of 82 adult and pediatric patients with the diagnosis of sinusitis with intracranial or orbital infection who were seen at the University of Mississippi Medical Center from January 1985 through December 1999. We assessed the diagnoses and outcomes to determine the most effective use of CT, with or without MRI, in the treatment of these patients. METHODS: Clinical presentations were reviewed, along with the CT and MRI findings, for patients who had sinusitis with complications. Clinical, CT, and MRI findings were compared for accuracy in diagnosing 43 patients with orbital infections and 39 patients with intracranial infections. RESULTS: For patients with orbital complications, most of whom had unilateral ethmoiditis, the diagnostic accuracy was 82% for clinical assessment compared with 91% for CT. For patients with intracranial complications, meningitis was the most common diagnosis, and MRI was more accurate (97%) in determining the diagnosis than CT (87%) or clinical findings (82%). CONCLUSION: CT remains the standard modality for diagnosing sinusitis, but MRI frequently is necessary, especially for patients with intracranial complications. Both diagnostic methods have improved the management and outcomes of patients who have sinusitis with complications. PMID- 11889375 TI - Use of AlloDerm for coverage of radial forearm free flap donor site. AB - OBJECTIVES: To evaluate and discuss the role of acellular human dermal matrix (AlloDerm, LifeCell Corp., Branchburg, NJ) graft for coverage of radial forearm free flap donor site. STUDY DESIGN: Prospective, nonrandomized study. METHODS: Fifty-two consecutive patients underwent harvest of 52 radial forearm fasciocutaneous free flaps. All 52 donor sites were covered by AlloDerm graft. Split-thickness skin graft was not used to reinforce the acellular dermal graft in the series. Clinical phases of healing, duration of healing, and donor site complications were studied. RESULTS: No donor site complications except seroma formation in five patients was noted. This was treated by conservative measures. Full range of hand motion was allowed in 3 days. Complete healing occurred within 8 to 12 weeks. Scar contracture after complete healing was minimum in all patients. Range of motion of the hand and fingers during flexion, extension, supination, and pronation was identical on the operated and nonoperated sides. CONCLUSION: AlloDerm graft is a viable alternative to split-thickness skin graft for coverage of the radial forearm free flap donor site. PMID- 11889376 TI - Recurrent attacks of facial nerve palsy as the presenting sign of leukemic relapse. AB - OBJECTIVE: To present an unusual case of recurrent facial palsy resulting from acute leukemic infiltration of the parotid gland. STUDY DESIGN: Case report. METHODS: An 11-year-old boy who had been treated for acute lymphoblastic leukemia (ALL) from 3 to 6 years of age presented with intermittent left facial nerve palsy with concurrent ipsilateral parotid fullness. The initial findings at diagnosis and workup are presented, and the disease progression and resolution with therapy are documented. RESULTS: The patient had been off therapy when this finding developed. A workup for central and viral etiologies for the facial palsy was unrevealing. Biopsy of the parotid gland demonstrated a lymphoblastic leukemic infiltrate. The patient was placed on a chemotherapy protocol for relapsed leukemia, resulting in complete resolution of the facial palsy. CONCLUSION: Isolated facial nerve dysfunction, albeit rare, has been documented as a sign of central nervous system involvement in leukemia, but until now this presentation has not been described in the setting of leukemic relapse presenting with acute infiltration of the parotid gland. PMID- 11889377 TI - Cyclooxygenase-2 expression in human thyroid carcinoma and Hashimoto's thyroiditis. AB - OBJECTIVES: Cyclooxygenases (COX) are enzymes that catalyze the conversion of arachidonic acid to prostaglandins. COX-2, unlike the constitutively expressed COX-1, is an inducible enzyme upregulated during cell proliferation and inflammation. More recently, COX-2 has been implicated in the development of numerous types of epithelial cancers. In addition, COX-2 is highly expressed in several inflammatory diseases. Because of its dual role in inflammation and cancer, we were interested in determining if COX-2 plays a role in the development of human thyroid carcinoma and Hashimoto's thyroiditis, an autoimmune condition frequently associated with thyroid malignancy. MATERIALS AND METHODS: Twenty paraffin-embedded human tissue specimens, including normal, inflammatory, and neoplastic thyroid sections, were analyzed by immunohistochemical staining for expression of human COX-2. In addition, COX-2 protein expression was verified by Western blot in two specimens. RESULTS: Immunohistochemical staining confirmed the presence of COX-2 in thyroid epithelial neoplasms, including papillary and follicular carcinomas. Moreover, COX-2 expression was observed in patients with Hashimoto's thyroiditis. COX-2 expression, however, was not observed in normal thyroid tissue, multinodular goiter, or anaplastic carcinoma. CONCLUSIONS: We have shown that cyclooxygenase-2 is expressed in thyroid carcinoma and thyroid epithelium from patients with Hashimoto's thyroiditis but not in normal thyroid. The expression of COX-2 in both of these thyroid pathologies may provide a basis for the relationship between carcinogenesis and autoimmunity. PMID- 11889378 TI - Topical mitomycin C and cephalosporin in endolymphatic sac surgery. AB - OBJECTIVE: To determine whether intraoperative topical application of the drug mitomycin C to the endolymphatic sac area in conjunction with endolymphatic sac ballooning surgery (ESBS) can safely achieve longer-lasting relief from incapacitating, medically intractable Meniere symptom-complex than ESBS alone. STUDY DESIGN/METHOD: A retrospective study is presented of 142 cases of ESBS performed by the author between May 1998 and November 1999, in which a solution of mitomycin C diluted to 1 mg/mL of physiological saline was applied to the sac area for 5 minutes followed by irrigation with a cephalosporin antibiotic solution. The results of the 103 of 142 cases diagnosed as classic Meniere's disease or endolymphatic hydrops are given special attention, comparing them with the results of an earlier-reported series of 109 cases of ESBS sharing the same diagnostic and result evaluation criteria but not using mitomycin C. RESULTS: In terms of vertigo control, the comparative short-term rates (<3 y) of complete/substantial control for the aforementioned 103-case series and earlier 109-case series are similar: respectively, 90.3% (after 1.5--3 years' follow-up) versus 91.7% (after 2.5-3 years' follow-up). In the 103-case series, to date, there has been no recurrence of incapacitating vertiginous symptoms necessitating revision surgery, whereas in the 109-case series, although there were likewise no cases necessitating revision surgery during the reported 2.5-to 3-year follow-up period, at least 8 patients in that series are known to have undergone revision surgery at later dates. Hearing results were significantly better (P =.004) in the 103-case series than the 109-case series: respectively, 30.0% versus 12.8% hearing improvement. CONCLUSIONS: Preliminarily, the superior hearing results in the present series indicate that possible adverse effects of mitomycin C on the inner ear and endolymphatic system have been averted; and no recurrence of incapacitating symptoms to date offers hope that longer-term efficacy of endolymphatic sac surgery using mitomycin C can be maintained. PMID- 11889379 TI - Tutorials in clinical research: part V: outcomes research. AB - OBJECTIVES: This is the fifth in a series of sequential "Tutorials in Clinical Research." The objective of the present report is to give the reader a broad overview of the field of outcomes research. This summary is intended to enable the reader to understand outcomes research methodology and to start the design of an outcomes research study. STUDY DESIGN: Tutorial. METHODS: The authors developed the report from available materials and refined it to be concise but complete for use by the practicing clinician. RESULTS: We describe the basic steps of record-based and patient-based outcomes research, including development of a staging system, identification of comorbid conditions, and creation or identification of an outcomes instrument. CONCLUSION: Outcomes research is a unique methodology that uses patient-based outcomes to assess the effectiveness of medical treatment. PMID- 11889380 TI - Successful cochlear implantation in prelingual profound deafness resulting from the common 233delC mutation of the GJB2 gene in the Japanese. AB - OBJECTIVES: Recently, we identified three novel mutations of the GJB2 gene in Japanese families with autosomal-recessive non-syndromic deafness.1 Seven of 11 mutated chromosomes (63.6%) contained a 233delC allele, suggesting that the 233delC mutation is the most common mutation of the GJB2 gene in the Japanese population. After it was recognized that cochlear implantation (CI) is of benefit to children with prelingual deafness, we have had a number of prelingual pediatric CI patients. Because children carrying the homozygous 233delC mutation show bilateral prelingual profound deafness, they could be enrolled in the CI program at Osaka University Graduate School of Medicine. The purposes of this study were 1) to analyze the occurrence of the GJB2 mutations in our 15 prelingual pediatric CI patients in whom the cause of non-syndromic deafness was unknown, and 2) to evaluate the auditory function and postoperative speech perception with CI of those GJB2-related deaf subjects. STUDY DESIGN: Retrospective analysis. METHODS: Mutation analysis of the GJB2 gene by direct sequencing was performed with genomic DNA from 15 children born profoundly deaf as a result of unknown causes and implanted with CI. Intraoperative electrically evoked auditory brainstem response (EABR) and intra-/postoperative EAP were measured. The speech perception was evaluated with Infants and Toddlers Meaningful Auditory Integration Scale (IT-MAIS). RESULTS AND CONCLUSIONS: We identified 4 CI patients (26.7%) out of 15 children carrying the homozygous 233delC mutation. Intra- and postoperative evaluation of the auditory system revealed almost intact cochlear and retrocochlear auditory function in these 4 patients. Postoperative auditory testing indicates that their speech perception had become significantly higher in comparison with that of other prelingual CI patients. These results suggest that prelingual deaf children carrying the homozygous 233delC mutation of the GJB2 gene can benefit from CI. PMID- 11889381 TI - The potential risk of carotid injury in cochlear implant surgery. AB - BACKGROUND: The advent of cochlear implantation has revolutionized the options afforded to the deaf population. With the increase in the prevalence of this procedure have come larger experiences in the associated technical challenges and complications. RESULTS: We present the evaluation and management of a patient with an unusual complication of improper placement of the implant electrode into the carotid canal and its management. We discuss the anatomy of the carotid artery and its proximity to the cochlea to emphasize the potential risk to this large vessel. CONCLUSIONS: Damage to the carotid canal and the carotid artery is a potential risk of cochlear implant surgery. When available, we recommend intraoperative electrical testing of the cochlear implant be performed. If there is doubt as to the placement of the electrode, a radiograph should be obtained before the patient is taken out of the operating room to avoid this complication. PMID- 11889382 TI - Preoperative versus postoperative role of vestibular-evoked myogenic potentials in cerebellopontine angle tumor. AB - OBJECTIVE/HYPOTHESIS: Vestibular-evoked myogenic potential (VEMP) examination was performed on patients with a cerebellopontine angle (CPA) tumor to evaluate its clinical role. METHODS: Patients with a CPA tumor were subjected to caloric test and VEMP examination. Follow-up study was performed 1 year after the surgery. RESULTS: Six (69%) of the 9 tumors did not exhibit either caloric response or VEMP on the lesioned side. Three patients received tumor excision and all tumors involved both the superior and inferior vestibular nerves. Two (22%) of the 9 tumors had normal caloric responses but no VEMP. One underwent surgical excision, and the tumor originated from the inferior vestibular nerve. In the follow-up study, only 1 patient with epidermoid cyst presented complete recovery of caloric response and VEMP, whereas in the other 3 patients with vestibular schwannoma, the responses were all absent persistently. CONCLUSION: Before surgery, VEMP test can be used to predict the nerve of origin and to formulate the best surgical approach. After surgery, VEMP test can be used to define the nature of the tumor (compressing or infiltrating the nerve) and disclose the residual function of the inferior vestibular nerve. PMID- 11889383 TI - Auditory manifestations of Keratitis-Ichthyosis-Deafness (KID) syndrome. AB - OBJECTIVE: Evaluation of the auditory manifestations of Keratitis-Ichthyosis Deafness (KID) syndrome, a rare genodermatosis characterized by follicular hyperkeratosis, vascularizing keratitis, and congenital hearing loss. STUDY DESIGN: Five individuals with sporadic KID syndrome were evaluated in the outpatient audiology clinic at the Warren Grant Magnuson Clinical Center of the National Institutes of Health. METHODS: Audiologic examinations included pure tone audiometry, speech audiometry, and middle ear immittance testing. Auditory brainstem responses and otoacoustic emissions were analyzed in 2 subjects. RESULTS: Four subjects had prelingual, bilateral, profound sensorineural hearing loss, whereas the fifth subject had significant residual hearing that exhibited no progression on serial audiograms. All 5 subjects had a history of non-erosive keratosis obturans and cutaneous cysts in the external ear canals that prevented continuous use of ear molds. CONCLUSIONS: The sensorineural hearing loss in KID syndrome is generally prelingual and profound. This combination of auditory and cutaneous phenotypes is similar to those previously reported for KID syndrome. KID syndrome presents a difficult challenge for communication rehabilitation because keratitis may impair the perception of sign and spoken language, and the cutaneous manifestations routinely curtail use of external amplification devices. PMID- 11889384 TI - Neonatal hearing loss in the indigent. AB - OBJECTIVE: To compare the risk factor profile for neonatal hearing loss (HL), and the follow-up rate of those identified with HL in an indigent population with those in an insured population. STUDY DESIGN: Retrospective review. METHODS: We studied 4526 neonates from the high-risk nursery or neonatal intensive care unit from two adjacent hospitals in Houston, Texas. Ben Taub General Hospital (BTGH) is a county public hospital that serves mainly the indigent. Texas Children's Hospital (TCH) is a private tertiary care center that serves patients with private insurance and Medicaid. RESULTS: Overall, 133 infants failed the screening test. Follow-up diagnostic testing identified 48 patients with definite HL. Although nearly twice as many patients at BTGH failed screening compared with TCH (88 vs. 45), four times as many patients at BTGH did not return for diagnostic testing (43 vs. 10). When a hearing aid was needed, there was a delay in getting one at BTGH (P <.05). There was a higher prevalence of dysmorphic facial features and central nervous system disease and a lower prevalence of long term ventilatory support at BTGH (P <.05). There were no differences between BTGH and TCH in the prevalence of low birth weight, neonatal asphyxia, syndromic stigmata, neonatal infection, family history of HL, or neonatal transfusion (P >.1). CONCLUSIONS: Significant differences in the risk factor profile for neonatal HL exist between the indigent and the general population. A worrisome problem exists with the timely intervention in hearing-impaired indigent neonates. PMID- 11889385 TI - Relationship between mastoid pneumatization and middle ear barotrauma in divers. AB - OBJECTIVES/HYPOTHESIS: Previous studies have shown a relationship between eustachian tube function and size of mastoid pneumatization, as well as eustachian tube function and middle ear (ME) barotrauma. The purpose of this study is to investigate a possible relationship between size of mastoid pneumatization and ME barotrauma in sports scuba (self-contained underwater breathing apparatus) divers. STUDY DESIGN: Prospective, blinded. MATERIAL AND METHODS: Twenty-four sports scuba divers (48 ears), who were fit to dive in the predive and otolaryngologic examination, were included in the study. Size of mastoid pneumatization was measured by simplified rectangular dimension method on a mastoid x-ray taken at Schuller's view. Divers were counseled to refer to the investigators if any symptoms occurred during and/or after diving. All symptomatic ears were examined within 24 hours of diving by the same investigator, who was blinded to the degree of pneumatization. RESULTS: ME barotrauma occurred in 15 ears (31%) of 11 divers (46%) at one time or another. The median degree of pneumatization in ears with barotrauma (22.9 cm2) was significantly smaller than that in unaffected ears (34.1 cm2; (P <.001). Furthermore, findings showed that with increasing degree of pneumatization, there was a decreasing risk of symptomatic barotrauma (P <.001). No barotrauma occurred in ears with a pneumatization greater than 34.7 cm2. However, barotrauma occurred in all 3 ears with a pneumatization degree smaller than 13.6 cm2. CONCLUSION: Our findings indicate an inverse relationship between size of pneumatization and risk of symptomatic ME barotrauma in sport scuba divers. PMID- 11889386 TI - Phenotype of DFNA11: a nonsyndromic hearing loss caused by a myosin VIIA mutation. AB - OBJECTIVES/HYPOTHESIS: To characterize the audiovestibular phenotype of DFNA11, an autosomal dominant nonsyndromic hearing impairment caused by a mutation in the myosin VIIA gene (MYO7A), including whether DFNA11-affected subjects have retinal degeneration as is characteristic of Usher syndrome type 1B, caused by different MYO7A mutations. STUDY DESIGN: Retrospective study of audiovestibular and ophthalmological data in a Japanese family linked to DFNA11. METHODS: Otoscopic examination and pure-tone audiometry were performed in all participants in the family. Selected subjects underwent additional examinations including speech discrimination scoring, acoustic reflex measurements, Bekesy audiometry, evoked and distortion-product otoacoustic emissions, auditory brainstem responses, and bithermal caloric testing; visual acuity, ocular tonometry, slit-lamp examination, ophthalmoscopy, and electroretinography; and computed tomography of the temporal bone. RESULTS: Most affected individuals had moderate cochlear hearing loss beginning in the second decade and progressing at all frequencies. Variable degrees of asymptomatic vestibular dysfunction were present. Computed tomography showed normal inner and middle ear structures. No evidence suggested retinitis pigmentosa. CONCLUSIONS: The phenotype of DFNA11 is postlingual, nonsyndromic sensorineural hearing loss with gradual progression. Showing moderate hearing loss with asymptomatic variable vestibular dysfunction and no retinal degeneration, the DFNA11 phenotype is mildest among phenotypes caused by MYO7A mutations. PMID- 11889387 TI - Spironolactone blocks glucocorticoid-mediated hearing preservation in autoimmune mice. AB - HYPOTHESIS: Although autoimmune sensorineural hearing loss can be effectively treated with corticosteroids, little is known about how these drugs affect cochlear function. MRL/MpJ-Faslpr autoimmune mice treated with a mineralocorticoid (aldosterone) have previously been shown to have hearing improvement equal to those treated with a glucocorticoid (prednisolone). This suggested that the restoration of hearing with steroids was the result of an effect on sodium transport rather than an antiinflammatory or immunosuppressive role. We hypothesized that corticosteroids reverse autoimmune hearing loss through the mineralocorticoid receptor and that blocking the mineralocorticoid receptor will prevent glucocorticoid effects. METHODS: Spironolactone, a mineralocorticoid receptor antagonist, was administered to MRL/MpJ-Faslpr autoimmune mice alone or in combination with corticosteroids. The four treatment groups were: spironolactone, spironolactone + aldosterone, spironolactone + prednisolone, and untreated water controls. Auditory brainstem response (ABR) thresholds were recorded before and during treatment (2, 3, and 4 mo) to measure the effect of steroids on hearing decline. RESULTS: Hearing in spironolactone and spironolactone + prednisolone mice showed progressive decline in hearing similar to water controls. The hearing was preserved in spironolactone + aldosterone mice, presumably as a result of the fact that aldosterone has a higher affinity for the mineralocorticoid receptor than spironolactone. Thus, aldosterone was able to maintain cochlear function with autoimmune disease progression, similar to previous reports of aldosterone treatment effects. CONCLUSIONS: Spironolactone effectively blocked prednisolone from improving hearing in MRL/MpJ-Faslpr autoimmune mice. This offers evidence that the inner ear mineralocorticoid receptor is the therapeutic target for corticosteroids used to treat autoimmune and sudden sensorineural hearing loss. Pharmacologic treatments that selectively target the mineralocorticoid receptor may provide greater clinical benefit with fewer systemic side effects than prednisone in patients with autoimmune sensorineural hearing loss. PMID- 11889388 TI - Validity of the Western blot immunoassay for heat shock protein-70 in associated and isolated immunorelated inner ear disease. AB - OBJECTIVE: To assess the validity of the Western blot immunoassay for heat shock protein-70 (hsp-70) for diagnosis of autoimmune inner ear disease. STUDY DESIGN: Retrospective study of 53 patients affected by sudden deafness (n = 19), idiopathic progressive sensorineural hearing loss (n = 24), and Meniere's disease (n = 10) who were treated from 1995 to 1999. The clinical course and response to corticosteroid were evaluated. METHODS: A purified hsp-70 antigen from bovine kidney cell line was used for the Western blot immunoassay. RESULTS: Only five patients (9.4%) showed anti--hsp-70 antibodies: Two presented a sudden sensorineural hearing loss (sudden deafness group), two showed an idiopathic progressive sensorineural hearing loss (idiopathic progressive sensorineural hearing loss group), and one was affected by fluctuating hearing loss (Meniere's disease group). A systemic autoimmune condition was observed in 29.1% of patients with idiopathic progressive sensorineural hearing loss. CONCLUSIONS: The low sensitivity of Western blot immunoassay for patients affected by idiopathic progressive sensorineural hearing loss and Meniere's disease may result from either the long time elapsed from the hearing loss and vertigo to the initial examination or from the increased percentage of cases of systemic autoimmune disease present in patients with idiopathic progressive sensorineural hearing loss. More studies to detect the immune-mediated inner ear disease in Western blot immunoassay-negative patients are required. PMID- 11889389 TI - Impact of functional endoscopic sinus surgery on symptoms and quality of life in chronic rhinosinusitis. AB - OBJECTIVES: Chronic rhinosinusitis restricts the quality of life of millions of involved patients. The aim of the study was to evaluate how functional endoscopic sinus surgery modifies patients symptom profiles and quality of life. STUDY DESIGN: Open prospective clinical trial. METHODS: Questionnaires were given to 279 patients included in the series, who underwent sinus surgery at the Department of Otorhinolaryngology--Head and Neck Surgery, University of Cologne (Cologne, Germany) from 1995 to 1999. Patients assessed typical chronic rhinosinusitis--associated symptoms and restricted quality of life preoperatively and postoperatively using ranking scales (scales ranging from no to intolerable complaints). Statistical analyses were performed with the Wilcoxon test and Spearman rank correlation coefficient. RESULTS: Quality of life was restricted by chronic rhinosinusitis in 94% of all patients preoperatively and ranked as severe or intolerable in 74%. Leading symptoms of chronic rhinosinusitis were nasal obstruction in 92% and postnasal drip in 87%. Furthermore, patients reported dry upper respiratory tract syndrome in 68%, hyposmia in 66%, headache in 64%, and asthmatic complaints in 34%. After a mean postoperative follow-up of 31.7 months, an amelioration of quality of life was achieved in 85%, no change in 12%, and a deterioration in 3%. The ranking of restricted quality of life improved from "severe" to "mild" (P <.01) in the mean. Mainly responsible for this improvement was the postoperative decrease of nasal obstruction (84%), headache (82%), and postnasal drip (78%) (all P <.01), which correlated significantly with nasal obstruction (r = 0.59), headache (r = 0.39), and postnasal drip (r = 0.55), respectively (all P <.01) with better quality of life. CONCLUSIONS: The leading complaints within the symptom profile of patients with chronic rhinosinusitis are airway obstruction and postnasal drip. The restriction of quality of life in patients with chronic rhinosinusitis is mainly caused by these symptoms, which can be improved in excellent fashion by functional endoscopic sinus surgery in the majority of patients, achieving better quality of life in the long term. PMID- 11889390 TI - Endoscopic repair of bilateral congenital choanal atresia. AB - OBJECTIVE: The literature about endoscopic repair of bilateral choanal atresia is scarce. The advantages and difficulties encountered with this technique are discussed. STUDY DESIGN: Prospective case series in a tertiary care center. METHODS: Nine infants with bilateral choanal atresia underwent transnasal endoscopic repair. On computed tomography scans, six had mixed atresia and three had bony atresia. Extra-long burrs, ear curettes, and dissectors all have been used with 4- and 2-mm, 0 degrees telescopes. The neochoana has been stented for 5 to 8 weeks. All cases were examined with the endoscope on removal of the stent; any granulation or polyps were removed at that time. RESULTS: Five cases remained patent after removal of stenting. Two patients required revision surgery because of repeat stenosis; one case remained patent and the other had repeat stenosis on one side. One infant died because of unrelated medical problems that occurred later. In one case the atretic tissue was thick, and the procedure was stopped because of bleeding; the infant died in the postoperative period after resuscitation. CONCLUSION: Careful review of the computed tomography scan and experience with endoscopic nasal surgery makes the transnasal endoscopic treatment a safe and effective approach for managing bilateral choanal atresia. PMID- 11889391 TI - The effects of topical agents of fluticasone propionate, oxymetazoline, and 3% and 0.9% sodium chloride solutions on mucociliary clearance in the therapy of acute bacterial rhinosinusitis in vivo. AB - OBJECTIVES: The aims of the study were to determine: 1) how mucociliary activity in acute bacterial rhinosinusitis is affected; 2) how this activity is changed by therapy; 3) the effects of topical agents on mucociliary clearance, and 4) the most appropriate topical agent(s) to be used in the therapy of sinusitis. STUDY DESIGN: Five groups of patients with acute bacterial rhinosinusitis were studied prospectively. METHODS: All patients had 500 mg oral amoxicillin and 125 mg oral clavulanic acid preparations given three times daily for 3 weeks. According to the topical agent applications, these groups included: group I (n = 12), no topical treatment was given; group II (n = 14), two puffs for each nostril once daily of 50 microg/100 mL fluticasone propionate was given; group III (n = 9), one puff for each nostril three times daily of 0.05% oxymetazoline was given; group IV (n =12), 3% sodium chloride (NaCl) (buffered to pH 6.5-7 at room temperature) was given; and group V (n =13), 10-mL solutions of 0.9% NaCl (buffered to pH 6.5--7 at room temperature) were given for nasal irrigations three times daily. All patients had medication for 3 weeks and were controlled each week. The saccharin method was used to measure nasal mucociliary clearance. To investigate the early effects of the topical agents for groups II to V, an additional test was repeated 20 minutes after the basal mucociliary clearance recordings. The test was repeated in the first, second, and third weeks of the treatment. RESULTS: The mucociliary clearance was significantly slower in the acute bacterial rhinosinusitis group than in the control group. There was no significant difference between the basal mucociliary clearance and the 20th minute mucociliary clearance of the fluticasone propionate and 0.9% NaCl solution groups. The mean values of the basal and the 20 minute's mucociliary clearance of the oxymetazoline group were 24.72 +/- 6.16 and 15.5 +/- 7.45 minutes, respectively, which were statistically significant. The mean values of the basal and the 20th minute mucociliary clearance of the 3% NaCl solution groups were 19.45 +/- 9.35 and 15.45 +/- 8.20 minutes, respectively, which were also statistically significant. In the first group (without topical treatment), the basal mucociliary clearance became significantly shorter after the second week of treatment. In the first and second weeks of the treatment of the oxymetazoline group, the mucociliary clearance did not change significantly, but after the third week the mucociliary clearance was significantly shorter. In the 3% NaCl solution group, significant improvement began from the first week and continued through the third week. Comparing the basal and the third weeks' mucociliary clearance values among the groups, the oxymetazoline and 3% NaCl solution groups revealed more significant improvement than the other groups, but this improvement was not different from the improvement of group I. There was still a statistically significant difference in the mucociliary clearance of the post treatment sinusitis groups from the control group. CONCLUSIONS: The oxymetazoline and 3% NaCl solution groups seemed to be more effective in mucociliary clearance, but there was no significant difference in improvement among the groups. The improvement of acute bacterial rhinosinusitis takes more than 3 weeks, according to the mucociliary clearance values of the groups. PMID- 11889392 TI - Mixed nasal mucus as a model for sinus mucin gene expression studies. AB - OBJECTIVE/HYPOTHESIS: It is necessary to obtain sinus mucus from the paranasal sinus cavities to study mucin gene expression occurring in the sinuses during chronic sinusitis. This requires an invasive procedure to access the sinus cavity. There are embryological as well as histological similarities between nasal and sinus epithelia; therefore, we postulated that the mucin expression in the secreted nasal and sinus mucins might be similar. Nasal mucus, which can be obtained easily, could then replace sinus mucus in these studies. STUDY DESIGN: Sinus and nasal mucus from six patients with chronic sinusitis were analyzed in this study. METHODS: High-molecular-weight glycoproteins (mucins) were isolated and purified by sequential density gradient centrifugation in caesium chloride (CsCl). Enzyme-linked immunosorbent assay was performed to identify the antigenic identity of these mucins. RESULTS: The MUC2, MUC5AC, and MUC5B mucin genes were all expressed in the nasal and sinus mucus secretions. Antigenic studies showed an inverse relationship between MUC2 and MUC5AC expression in nasal and sinus mucus secretions. The MUC5B gene was the major mucin gene expressed in sinus mucus but not in nasal mucus. Expression of MUC2 was significantly higher in sinus mucus. Expression of MUC5AC was different between nasal and sinus mucus. CONCLUSIONS: Individual mucin expression in sinus and nasal mucus was markedly different. From this preliminary study, we conclude that nasal mucus is not a suitable substitute for sinus mucus in sinus mucin gene studies and that different pathological processes are taking place in nasal and sinus tissue in chronic sinusitis. PMID- 11889393 TI - Videofluoroscopic upper esophageal sphincter function in elderly dysphagic patients. AB - OBJECTIVES/HYPOTHESIS: The intent of the study was to identify and characterize abnormalities of the timing and extent of upper esophageal sphincter (UES) opening in an elderly population complaining of dysphagia. STUDY DESIGN: A retrospective review of dynamic swallow studies performed on patients greater than 65 years of age without an obvious medical or surgical cause for their dysphagia. METHODS: Measures of UES opening timing and extent in the patient population were compared with those from 60 young, normal control subjects and 23 elderly control subjects. The relationship of UES function and other swallowing abnormalities was also evaluated. RESULTS: No decrease in the size of UES opening was identified in the patient population. The coordination of UES opening relative to the position of the bolus in the pharynx was normal. UES opening was prolonged and was correlated with poor pharyngeal clearing suggestive of weak pharyngeal constriction. CONCLUSION: No primary abnormality of UES function was identified in this elderly dysphagic patient population. PMID- 11889394 TI - Laryngeal adductor reflex and pharyngeal squeeze as predictors of laryngeal penetration and aspiration. AB - OBJECTIVES: The contribution of laryngopharyngeal (LP) sensory deficits to the outcome of swallowing and the relationship between sensory and motor deficits in the laryngopharynx is unclear. The study purpose is to determine if patients with LP sensory and motor deficits are at increased risk for laryngeal penetration and aspiration during swallowing, and to determine the relationship between pharyngeal motor weakness and LP sensory deficits. MATERIALS AND METHODS: Endoscopic evaluation of swallowing with sensory testing was performed on 122 dysphagic patients who were prospectively divided into two groups. The control group was 76 patients with normal sensitivity, determined by an intact laryngeal adductor reflex (LAR) on air pulse stimulation of the mucosa innervated by the superior laryngeal nerve. The study group was 46 patients with severe sensory deficits, determined by an absent LAR. Each group was given puree followed by thin liquid, noting presence or absence of laryngeal penetration and aspiration. Pharyngeal muscle strength was assessed by noting presence or absence of pharyngeal contraction during voluntary adduction of the vocal folds (pharyngeal squeeze). RESULTS: In control subjects, with purees, 6 of 76 (7.90%) penetrated and 3 of 76 (3.94%) aspirated; with thins, 26 of 76 (34.2%) penetrated and 13 of 76 (17.1%) aspirated. In the absent LAR group, with purees, 39 of 46 (84.8%) penetrated and 32 and 46 (69.6%) aspirated; with thins, 46 of 46 (100%) penetrated and 43 of 46 (93.5%) aspirated. For both consistencies, the differences in prevalence of penetration and aspiration between groups was significant (P <.0001, chi2). In control subjects, pharyngeal squeeze was impaired in 17 of 76 (22.4%), with penetration of puree in 6 of 17 (35.3%) and aspiration in 3 of 17 (17.6%). In the absent LAR group, squeeze was impaired in 41 of 46 (89.1%), with penetration of puree in 39 of 41 (95.1%) and aspiration in 32 of 41 (78.0%). The difference in the prevalence of pharyngeal weakness between groups was significant (P <.0001). The difference in the prevalence of penetration and aspiration was higher in the absent LAR/impaired contraction cohort than in the normal sensation/impaired contraction cohort (P <.0001). CONCLUSION: Absence of the LAR and impaired pharyngeal squeeze puts patients with dysphagia at high risk for laryngeal penetration and aspiration compared with patients with an intact LAR and intact pharyngeal squeeze. There is a strong association between motor and sensory deficits in the laryngopharynx. PMID- 11889395 TI - A new paramedian approach to arytenoid adduction and strap muscle transposition for vocal fold medialization. AB - OBJECTIVE: To develop a prosthesis-free medialization laryngoplasty for the treatment of glottal incompetence. STUDY DESIGN: Twenty-two consecutive patients with glottal incompetence underwent vocal fold medialization using a new paramedian approach to arytenoid adduction and/or strap muscle transposition. METHODS: Under local anesthesia, the thyroid lamina on the involved side was parasagittally separated 5 mm off the midline. The inner perichondrium was carefully freed from the overlying thyroid cartilage. After dividing the thyrohyoid and cricothyroid membranes, the lamina was retracted laterally, the inner perichondrium was opened, and the lateral cricoarytenoid muscle identified. Tracing the muscle fibers posterosuperiorly, the muscular process of the arytenoid was identified. A 2-0 or 3-0 Prolene suture was placed through the muscular process and tied to the cricoid cartilage at the origin of the lateral cricoarytenoid muscle. A bipedicled strap muscle flap was then transposed into the space between the lamina and the inner perichondrium and the thyroid cartilages sutured back into place. Pre- and postoperative voice evaluations measured mean fundamental frequency, jitter, shimmer, noise-to-harmonic ratio, and maximal phonation time, as well as assessments of voice quality. RESULTS: Vocal improvement was obtained in 95% (21 of 22) of patients. There was a significant improvement (P <.05) in all parameters except shimmer. No major complications were noted in any patient, except for dyspnea in one patient resulting from arytenoid overrotation. CONCLUSION: The results suggest that a paramedian approach to arytenoid adduction combined with strap muscle transposition is a safe and effective method for treating glottal incompetence, particularly in patients with unilateral paralytic dysphonia. PMID- 11889396 TI - The effect of neuromuscular stimulation of the genioglossus on the hypopharyngeal airway. AB - OBJECTIVES: To determine the effects of neuromuscular stimulation (NS) of the genioglossus muscle on hypopharyngeal airway size. STUDY DESIGN: Fourteen consecutively recruited healthy volunteers underwent percutaneous electrical NS of the genioglossus muscle. METHODS: Bipolar hooked wires were inserted percutaneously into the genioglossus muscle and used for NS. The anterior- posterior diameter of the hypopharynx was measured at the level of the superior edge of the epiglottis at baseline and during NS from recorded video endoscopic examinations. RESULTS: NS of the genioglossus muscle resulted in a significant increase in the diameter of the hypopharyngeal airway (P =.002) compared with baseline, ranging from a 33% to 284% increase in airway diameter. Three of the 14 patients demonstrated modest decreases in airway diameter, likely the result of faulty electrode placement in surrounding tongue retrusive muscles. CONCLUSIONS: NS of the genioglossus muscle was effective in increasing the hypopharyngeal airway and may provide a useful alternative to direct stimulation of the hypoglossal nerve with a nerve cuff electrode in the development of neuroprosthetic treatments for obstructive sleep apnea. PMID- 11889397 TI - Histopathology of the uvula and the soft palate in patients with mild, moderate, and severe obstructive sleep apnea. AB - OBJECTIVE: To study morphometric and qualitative histopathologic changes of the soft palate and uvula in patients with mild, moderate, and severe obstructive sleep apnea. STUDY DESIGN: A prospective, nonrandomized controlled study. METHODS: The distal soft palate and uvula were excised during uvulopalatopharyngoplasty from 34 male patients with obstructive sleep apnea. Control specimens were retrieved from 7 male cadavers with no related disorders. All specimens underwent routine processing and the mid-sagittal sections were studied. Morphometric analysis of the relative proportions of the tissue constituents was carried out. Also, a qualitative assessment was performed to detect possible pathologic changes. RESULTS: The body mass index of patients was significantly higher from that of control subjects. The area fraction occupied by the tissue constituents of the distal portion of the soft palate and uvula in patients with mild, moderate, and severe obstructive sleep apnea and in control subjects was similar, with small and insignificant differences regarding the contents of glands, muscle, fat, blood vessels, and the epithelium. Only the connective tissue was significantly greater in patients with moderate obstructive sleep apnea than in those with severe obstructive sleep apnea and control subjects. The qualitative assessment of the specimens disclosed normal tissue architecture without evidence of destruction. Vascular engorgement, fibrosis, edema, inflammatory cell infiltration, and dilated glandular ducts were observed in a portion of patients and control subjects. CONCLUSIONS: The structure of the distal soft palate and uvula of patients with obstructive sleep apnea undergoes insignificant changes and is independent of the body mass index levels, indicating that the pathologic changes are probably the sequela of airway obstruction rather than its cause. PMID- 11889398 TI - The mechanical stability under load of tracheal anastomoses after various phases in vivo. AB - OBJECTIVE: The objective of our study was to postoperatively examine the mechanical stability under load of tracheal anastomoses over different periods in time and to compare these with native tracheae. STUDY DESIGN: A randomized experimental study on animals. METHOD: We performed tracheal anastomoses on sheep with three different suturing techniques and different resection lengths (3, 6, and 9 cm): These anastomoses were subjected to a breaking test at different intervals in vivo (1, 2, 4, 8, and 24 wk) and compared with the tracheae of healthy sheep. RESULTS: After 1 week in vivo, the anastomosis itself tore off from the remaining trachea under tension, regardless of the suturing technique used and the length of resection. With all animals that survived for a longer period, the trachea broke a greater distance away from the anastomosis. The necessary breaking forces were only minimally lower than those required for breaking healthy tracheae and the difference is statistically insignificant. When all operated tracheae are combined and the forces compared with native tracheae, this reveals that the operated tracheae are significantly more stable (P =.015) and present a lower longitudinal elasticity (P =.004). CONCLUSION: During the first postoperative days, the stability under load of tracheal anastomoses is slightly lower than that of healthy trachea. This difference is, however, far from those values that can be measured intraoperatively on tracheal anastomoses. Thus, supplementary measures for the mechanical protection against suture line separation do not seem necessary. PMID- 11889399 TI - CO2 laser cordectomy for early-stage glottic carcinoma: a long-term follow-up of 156 cases. AB - OBJECTIVES: To define when laser resection of early-stage glottic carcinoma is indicated and to compare the results obtained by laser surgery with other therapeutic options. STUDY DESIGN: Retrospective study of 151 patients treated from April 1982 to June 1996 in the Department of Otorhinolaryngology at "La Sapienza" University. We provide analysis of indications, techniques, and oncologic results of this study. METHODS: Glottic tumors were treated with type III, type IV, and type Va cordectomies according to the classification of endoscopic cordectomies proposed by the European Laryngological Society in 2000. RESULTS: The results are summarized as follows: all patients with carcinoma in situ Tis are free of disease with local control rate at 3 years of 100%; 2 died of other causes without evidence of local recurrence with an overall survival rate at 3 years of 83.2%. Of the 117 patients with stage T1a cancer, 110 are free of disease at 3 years with local control rate of 94%; 4 patients died of other causes without evidence of local recurrence with an overall survival rate of 96.5%. Of the 22 patients with stage T1b cancer, 20 are free of disease at 3 years with a local control rate of 91%; 1 patient died of other causes without evidence of local recurrence with an overall survival rate at 3 years of 95.4%. CONCLUSIONS: According to our experience, we can conclude that endoscopic laser surgery is an efficacious and cost-effective treatment for early stage glottic cancer. PMID- 11889400 TI - Near-total laryngectomy in advanced laryngeal and pyriform cancers. AB - OBJECTIVE: To demonstrate the oncologic and physiological safety of near-total laryngectomy (NTL), its success in voice conservation, and its versatility for use in extensive resections that necessitate pharyngoplasty, and even in post radiation recurrences. STUDY: In this study of 137 cases of NTL for cancer of the larynx (45 cases) and pyriform (92 cases), 86.9% were stage T3/T4 and 60.6% were N+. A total of 8.8% had extended pharyngeal resections necessitating patch pharyngoplasty (ENTLP). In 10.9% cases, NTL was used as salvage of post-radiation failures. Concurrent neck dissection was performed in 99 cases. RESULTS: A total of 70.1% was alive and disease-free at the last follow-up ranging from 12 months to 104 months (median, 35 mo). A total of 7.3% had local/locoregional recurrences and 11.7% had purely regional recurrences. The local control rate for post radiation salvage with NTL was 93.3%. A total of 88.6% developed communicable speech, and the speech success rate was 100% in 12 cases of ENTLP. Complications included major wound dehiscence with total shunt breakdown in 2 cases (1.5%), pharyngeal leak requiring surgical intervention in 7 cases (3.6%), significant aspiration through the shunt necessitating completion laryngectomy in 1 case (0.7%), and complete shunt stenosis in 9 cases (6.6%). CONCLUSION: The study shows that NTL is an oncologically safe voice conservation procedure in advanced, lateralized laryngeal and pyriform cancers treated not only per primum, but also in carefully selected post-radiation failures. It has a high success rate of speech development even in those cases requiring extensive pharyngeal resections. Major complications were acceptably low. PMID- 11889401 TI - FDG PET for mucosal malignant melanoma of the head and neck. AB - OBJECTIVES/HYPOTHESIS: To evaluate imaging findings using positron emission tomography (PET) and 18F-fluorodeoxyglucose (FDG) in mucosal malignant melanoma (MMM) of the head and neck. STUDY DESIGN: Retrospective evaluation. METHODS: Eighteen PET examinations were performed for initial staging and/or follow-up in 10 patients with MMM. Medical records of 6 male and 4 female patients (age range, 43--81 y; mean, 67 y) were reviewed retrospectively with regard to the patients' history, symptoms, and clinical course. Primary melanoma elsewhere in the body was excluded at the time of diagnosis. RESULTS: All MMM were visible in staging PET examinations, but FDG uptake depended on lesion size and anatomic site. Big lesions with a nodular growth, as seen in pathologic specimens, were better visible compared with lesions with a more superficial spread within the mucous membranes. Lesions in the anterior part of the nasal cavity were more difficult to detect than those in the posterior sinonasal complex because of possible interference with nonspecific uptake in muscles of the mouth and pronounced appearance of the skin when imaging was performed using filtered back projection without attenuation correction. CONCLUSION: We found that MMM of the head and neck can be visualized using FDG PET. Furthermore, locoregional and distant metastases can be evaluated much like those of cutaneous malignant melanoma. Therefore, PET may be suitable for the staging and/or restaging of these patients. Further studies have to elucidate the potential role of FDG PET in patient management. PMID- 11889402 TI - Microsurgical anatomy of the laryngeal nerves as related to thyroid surgery. AB - OBJECTIVES: The objectives were to explore microsurgical anatomy of the superior and recurrent laryngeal nerves and their importance in thyroid surgery, and to examine areas of potential morbidity, means of identification, and arterial supply of the laryngeal nerves. STUDY DESIGN: Descriptive analysis of anatomical features. METHODS: Twenty-one adult cadavers, some perfused with colored silicon, were dissected for the study project. RESULTS: The right recurrent laryngeal nerve (RLN) branches off the vagus at the level of the subclavian artery and the left one at the level of the aorta. Both ascend parallel to the tracheoesophageal groove and innervate trachea, esophagus, and the inferior pharyngeal constrictors en route. The RLN has the highest probability to pass between the branches of the inferior thyroid artery on the right side and posterior to them on the left side. The RLN always passes posterior to the cricothyroid joint. The RLN is supplied by the branches of the inferior thyroid artery. The superior laryngeal nerve (SLN) branches into internal and external branches deep to the carotid bifurcation. The internal branch passes deep to the superior thyroid artery and descends toward thyrohyoid membrane. The external branch travels deep and parallel to the superior thyroid artery to innervate cricothyroid muscle. The internal branch is supplied by the superior laryngeal artery, and the external branch by the cricothyroid artery. CONCLUSIONS: The only consistent location of the RLN is when it passes posterior to the cricothyroid joint. Because of extreme variability of the inferior thyroid artery and the RLN, it is suggested that the artery be ligated either proximally or at its tertiary branches on thyroid capsule. The internal branch of the SLN is not potentially at risk during thyroidectomy unless the superior thyroid artery is ligated proximally. The external branch of the SLN accompanies the superior thyroid artery for most of its course and is at potential risk if the trunk of the superior thyroid artery is ligated outside the pretracheal fascia. PMID- 11889403 TI - Glut3 expression in biopsy specimens of laryngeal carcinoma is associated with poor survival. AB - OBJECTIVES/HYPOTHESIS: The aim of the study was to determine the clinical significance of the expression of Glut1 and Glut3 proteins in biopsy specimens of squamous cell carcinoma (SCC) of the larynx. STUDY DESIGN: A retrospective study. METHODS: Using immunohistochemistry, we immunostained sections of formalin-fixed, paraffin-embedded tissues from 48 biopsies of invasive SCC of the larynx for Glut1 and Glut3. The percentages of positive cells were recorded, then correlated with overall patient survival using the Kaplan-Meier method and the Breslow-Gehan Wilcoxon test for statistical significance. RESULTS: All cases were positive for Glut1, and Glut1 expression was not associated with survival difference at any cut-off value. Eighteen (38%) of the cases were Glut3-negative and 30 (62%) were Glut3-positive. Glut3-positive cases were associated with poorer survival than Glut3-negative cases (P =.0336). No significant difference was found between Glut3-negative and Glut3-positive groups in respect to sex, tumor site (glottic vs. supraglottic), nodal or distant metastasis, or treatment modality. However, there were significantly more poorly differentiated tumors in the Glut3-positive group than in the Glut3-negative group (27% vs. 0%, respectively; P =.0182, Fisher's Exact Test). After poorly differentiated tumors were excluded from the survival analysis, Glut3 immunoreactivity remained a significant marker of poor prognosis (P =.0385). CONCLUSION: Immunohistochemical detection of Glut3 in biopsy specimens of SCC of the larynx is a marker of poorer prognosis. PMID- 11889404 TI - First evidence of genetic imbalances in angiofibromas. AB - OBJECTIVE/HYPOTHESIS: Angiofibromas are clinically well characterized by their origin at the posterior lateral nasal wall close to the sphenopalatine foramen, their occurrence in male adolescent patients, and the histological findings of a benign fibrovascular neoplasm with irregular, endothelium-lined vascular spaces in a fibrous stroma. However, their etiology and genetic causes remain unknown. The present study addresses genetic imbalances in angiofibromas. STUDY DESIGN: The present pilot study compared genomic hybridization in three angiofibromas to search for chromosomal abnormalities in this rare tumor. METHODS: Fluorescence marked normal DNA and angiofibroma DNA were compared using genomic hybridization screening to detect chromosomal abnormalities. Their binding ratio to metaphase chromosomes were analyzed by special digital image analysis. RESULTS: Chromosomal gains and losses showing a high level of agreement were detected in all three angiofibromas. Specifically, DNA gains were observed on chromosomes 3q, 4q, 5q, 6q, 7q, 8q, 12p, 12q, 13q, 14q, 18q, 21q, and X, and DNA losses were screened on chromosomes 17, 19p, 22q, and Y. Finding chromosomal abnormalities at the sex chromosomes X and Y of this rare tumor is remarkable. Concurrent chromosomal gain on 8q12q22 was noted in all three tumor specimens. CONCLUSIONS: Comparative genomic hybridization is suitable for screening angiofibromas on a genetic level. The results on these screens indicate that further genetic investigations of this rare benign tumor may provide more details about the tumor's genetic abnormalities and perhaps clarify the etiology of angiofibromas. PMID- 11889408 TI - Economic analysis. PMID- 11889405 TI - Association between high initial tissue levels of cyclin d1 and recurrence of nasopharyngeal carcinoma. AB - OBJECTIVES/HYPOTHESIS: Cyclin D1 expression and the rate of apoptosis have been reported to serve as important prognostic indicators in human cancers. The purpose of the present study was to determine the prognostic significance of both initial cyclin D1 expression and the apoptotic index in nasopharyngeal carcinoma. STUDY DESIGN: Cohort study. METHODS: Cyclin D1 protein levels and apoptosis in tumors and their corresponding adjacent, histologically normal tissues were determined at the time of initial diagnosis using immunohistochemical staining, Western blot analysis, and in situ end labeling, respectively, in 64 patients with T1-T4/N0-N2, poorly differentiated squamous cell carcinoma of the nasopharynx. All cases were treated by routine radiation therapy with a total median dose of 70 Gy and followed up for 10 years. RESULTS: High levels of cyclin D1 were found in 35 of 64 tumor specimens (54.7%); no cyclin D1--positive cells were determinable in normal epithelium of the nasopharynx. Rates of early local recurrence (within 5 y) were significantly higher (P <.01) for patients with high levels of cyclin D1 before radiation therapy (24 of 35 patients [68.6%]) as compared with patients with low or no expression (3 of 29 [10.3%]). Furthermore, patients bearing high levels of cyclin D1 had a poorer prognosis concerning 10 year survival than the others (P <.001), whereas overexpression of cyclin D1 did not correlate with the initial TMN classification (P >.05). According to the rate of spontaneous apoptosis in tumors below or above the median, patients were divided into two groups. There was no statistically significant difference for the overall survival between the two groups (P >.05). CONCLUSIONS: The present study demonstrates that cyclin D1 can be used as an indicator of recurrence and subsequent prognosis in nasopharyngeal carcinoma after radiation therapy. At the same time, the apoptotic state before radiation therapy is of no value in predicting the prognosis of patients with nasopharyngeal carcinoma. PMID- 11889409 TI - An aid to accurate skin closure. PMID- 11889410 TI - Clinical course of the syndrome of autoantibodies to the insulin receptor (type B insulin resistance): a 28-year perspective. PMID- 11889411 TI - Familial expansile osteolysis (excessive RANK effect) in a 5-generation American kindred. PMID- 11889412 TI - Natural history of Fabry renal disease: influence of alpha-galactosidase A activity and genetic mutations on clinical course. PMID- 11889413 TI - Systemic sclerosis: demographic, clinical, and serologic features and survival in 1,012 Italian patients. PMID- 11889414 TI - Predicting mortality in systemic sclerosis: analysis of a cohort of 309 French Canadian patients with emphasis on features at diagnosis as predictive factors for survival. PMID- 11889415 TI - A guest editorial: where are we now in screening patients for preterm delivery? PMID- 11889416 TI - Topical anesthesia for minor gynecological procedures: a review. AB - This article reviews the published literature for topical anesthetics that have been used for pain relief during minor gynecological procedures. EMLA (an eutectic mixture of the local anesthetics, lidocaine 2.5% and prilocaine 2.5%), which is the best-studied topical anesthetic, produces effective analgesia for superficial surgical procedures after application for 5 to 10 minutes and has been extensively studied in various procedures including removal of genital warts, vulval biopsy, laser treatment of CIN lesions, and hysteroscopy. EMLA is well tolerated and provides good pain relief for procedures involving the surface tissues such as removal of genital warts and hysteroscopy. For procedures involving deeper tissues, EMLA reduces the pain of local anesthetic injection. Other topical anesthetics, such as lidocaine gel and spray, benzocaine 20% gel, mepivacaine solution, tetracaine solution, and cocaine spray, have been less extensively studied in these indications, and benefits seem to be limited. PMID- 11889417 TI - Headache during pregnancy. AB - Headache in pregnancy is a common problem but diagnosis and management can be challenging. Most headaches in pregnant women are either migraine or tension types and can be easily treated. Rarely, the headache in a pregnant patient signals a life-threatening condition. Obstetricians should be able to effectively manage the common causes of headache as well as recognize the warning signs of potentially serious conditions. Diagnosis and management should be systematic yet individualized. PMID- 11889418 TI - The role of mitochondria in ischemia/reperfusion injury. AB - In organ transplantation, ischemia/reperfusion injury is a multifactorial process that leads to organ damage and primary graft dysfunction. Injury to the organ is mediated by a complex chain of events that involves depletion of energy substrates, alteration of ionic homeostasis, production of reactive oxygen species, and cell death by apoptosis and necrosis. There is increasing evidence that mitochondria play a role in this process because of the profound changes experienced during ischemia and reperfusion. Understanding the mechanisms that lead to mitochondrial damage may be important for developing strategies aimed at improving graft outcome. In this review, we examine the role of mitochondria in ischemia/reperfusion injury and the possible mechanisms that may contribute to organ dysfunction. PMID- 11889419 TI - Prevention of bronchiolitis obliterans in rat lung allografts by type V collagen induced oral tolerance. AB - BACKGROUND: We have reported that feeding type V collagen (col(V)) to lung allograft recipients induces immune tolerance that prevents acute lung allograft rejection. Repeated acute rejection is a risk factor for or associated with chronic rejection, known as bronchiolitis obliterans (BO), the leading cause of death in lung allograft recipients. The current study examines if col(V)-induced oral tolerance prevents BO. METHODS: WKY rats (RT1l) were fed either col(V) or diluent before orthotopic transplantation of F344 (RT1lvl) lung allografts. No rats received any immunosuppression. At 10 weeks posttransplantation the time to onset of BO, delayed type hypersensitivity (DTH) responses to donor antigens, and col(V) were examined. In addition, proliferative responses of recipient T lymphocytes to donor antigens, and ability of recipient antigen presenting cells to present alloantigens in lung allografts were evaluated. RESULTS: The data show that recipient rats have sustained DTH responses to donor antigens and col(V). T lymphocytes from col(V)-fed lung allograft recipients were unable to proliferate in response to donor antigens, but feeding col(V) had no effect on the presentation of donor alloantigens by recipient antigen presenting cells. All diluent fed rats developed BO, but only mild acute rejection (grade 2) was present in all rats fed col(V). Transforming growth factor (TGF)-beta production was up-regulated systemically in col(V)-fed, but not diluent fed, lung allograft recipients, and neutralizing TGF-beta [corrected] recovered the DTH response to donor antigens in col(V)-fed rats. CONCLUSIONS: Collectively these data show that col(V)-induces oral tolerance that prevents BO, and that tolerance may be mediated by systemic production of TGF-beta [corrected]. PMID- 11889420 TI - The effect of acute rejection and cyclosporin A-treatment on induction of platelet-derived growth factor and its receptors during the development of chronic rat renal allograft rejection. AB - BACKGROUND: In the development of chronic kidney allograft rejection acute rejection (AR) is the single most important risk factor. Although Cyclosporin A (CsA) medication has decreased the incidence of AR, chronic rejection (CR) is still the major reason for late allograft loss. Platelet-derived growth factor (PDGF) is a major mitogen mediating mesenchymal cell proliferation in CR. We have investigated the impact of AR and different doses of CsA on the expression of PDGF ligands and receptors in the development of CR. METHODS: Kidney transplantations were performed from DA to WF rats and syngenic controls were done from DA to DA rats. Two groups of allografts were treated daily with CsA either at low dose (1.5 mg/kg) or high dose (5 mg/kg). Third group of allografts was treated with CsA 5 mg/kg/day for 1 week and then left untreated until the development of AR. AR episodes were treated with CsA 5 mg/kg/day. Grafts were harvested 3 months after transplantation for histology and immunohistochemistry (PDGF-AA, -BB and PDGFR-alpha, -beta). RESULTS: In syngenic grafts no histological signs of CR were seen and the expression of PDGF ligands and receptors remained almost nonexistent. AR episodes increased the chronic rejection changes. High-dose CsA-treatment ameliorated inflammation compared to low-dose CsA-treatment, although it failed to inhibit the development of chronic changes. More fibrosis was even seen in high-dose than in low-dose CsA-treated grafts. CR in each allograft group was associated with induction of all PDGF ligands and receptors (P<0.05 compared with syngenic controls) in interstitial inflammatory cells, capillary endothelium, and arterial smooth muscle cells. In the group with AR episodes the expression was further increased. CONCLUSIONS: Our results demonstrate that CsA treatment cannot inhibit the expression of PDGF ligands and receptors in the development of chronic kidney allograft rejection and that AR episodes induce even more PDGF and its receptors in the graft indicating a link between AR and subsequent development of CR. PMID- 11889421 TI - Treatment of streptozotocin-induced diabetes mellitus by transplantation of islet cells plus bone marrow cells via portal vein in rats. AB - BACKGROUND: We have established a new method for the transplantation of allogeneic pancreatic islets (PIs) using sublethal irradiation (9 Gy) plus simultaneous transplantation of PIs and bone marrow cells (BMCs) via the portal vein (PV) followed by intravenous (i.v.) injection of donor BMCs (9 Gy + PV + i.v.). METHODS: Approximately 600 PIs of Brown Norway (BN: RT1An, RT1Bn) rats were transplanted into the liver of streptozotocin-induced diabetic Fischer 344 (F344: RT1Al, RT1Bl) rats via the PV. BMCs (3x108) of BN rats were injected via the PV or i.v. into the recipients simultaneously. In some groups, additional i.v. injections of BMCs from BN rats were given 5 days after the PI transplantation. RESULTS: All the recipients (10 of 10) in the 9 Gy + PV + i.v. group showed normoglycemia for more than 1 year, whereas PIs were rejected within 30 days after transplantation in the group of 9 Gy + i.v. + i.v. CONCLUSIONS: These results suggest that simultaneous transplantation of PIs and BMCs via the PV is effective in inducing persistent tolerance. PMID- 11889422 TI - Survival of fetal skin grafts is prolonged on the human peripheral blood lymphocyte reconstituted-severe combined immunodeficient mouse/skin allograft model. AB - BACKGROUND: Fetal tissue is considered to be immune privileged and is under extensive investigation as a source of tissue for transplantation. In this paper, we analyzed the immune properties of human fetal and neonatal skin before and after transplantation to severe combined immunodeficient (SCID) mice. Using a human peripheral blood mononuclear cell reconstituted SCID (huPBMC-SCID) mouse model of allograft rejection, we compared the immune response to transplanted fetal and neonatal skin. METHODS: We analyzed human fetal (55-122 days of gestation) and neonatal skin samples by routine histology and immunohistochemistry for the expression of (MHC class I and II antigens before and after transplantation to SCID mice. After transplantation, we injected the mice with huPBMCs and analyzed the survival of neonatal and fetal skin grafts both visually and microscopically. RESULTS: We detected no class II expression in fetal skin of all gestational ages and only weak class I expression after 89 days compared with abundant class I and II expression in neonatal skin before transplantation. When transplanted to SCID mice, fetal skin grafts differentiated and expressed class I and II, but the levels were lower than neonatal grafts. In mice injected with huPBMCs, rejection of neonatal grafts started on day 5, and by day 9 all grafts were rejected. In contrast, rejection of fetal skin grafts was significantly delayed. Rejection started on day 13 and was complete by day 23 (P<0.00005). Histology sections from the rejected grafts showed marked CD3+ T cell infiltration in the human skin with a sharp demarcation between the human and mouse skin, with no T-cell infiltration in the mouse skin. CD4+ and CD8+ T cells were present in the rejected sites in similar densities. CONCLUSIONS: Our results show that fetal skin differentiates and expresses increased amounts of MHC class I and class II antigens when transplanted to SCID mice. However, these levels are much lower than the levels found in neonatal skin. We demonstrate that the survival of human fetal skin allografts is markedly prolonged compared with that of neonatal skin grafts in the huPBMC-SCID mouse model. Our results support the hypothesis that low levels of MHC antigen expression lead to a delay in the rejection of fetal skin and further demonstrate the utility of the huPBMC-SCID mouse model to investigate the molecular and cellular mechanisms of the immune response to human fetal tissues. PMID- 11889423 TI - TCV-116, an angiotensin II type 1 receptor antagonist, reduces hepatic ischemia reperfusion injury in rats. AB - BACKGROUND: In Pringle's maneuver during liver surgery and liver transplantation, ischemia-reperfusion (I/R) is an unavoidable process, and protection against hepatic I/R injury is a major unresolved problem. Therefore, various pharmacologic approaches to prevent hepatic I/R injury are currently under trial. In this study, we investigated whether TCV-116, an angiotensin II type 1 receptor antagonist, can reduce this injury. METHODS: The rats were pretreated either with TCV-116 (group 1) or with the vehicle alone (group 2). The rats in group 3 were not pretreated. Thereafter, they were subjected to partial hepatic I/R. RESULTS: After reperfusion, the mean peak plasma concentrations of aspartate aminotransferase, alanine aminotransferase, lactic dehydrogenase, and creatine kinase were lower in group 1 than in groups 2 and 3. The magnitude of hepatic injury was reduced in group 1 compared with that in groups 2 and 3. The mean peak plasma concentrations of tumor necrosis factor-alpha, cytokine-induced neutrophil chemoattractants-1, and interleukin-6 were lower in group 1 than in groups 2 and 3. The number of neutrophils infiltrating the liver was also lower in group 1 than in groups 2 and 3. The mean peak plasma concentration of hepatocyte growth factor (HGF) was higher in group 1 than in groups 2 and 3. CONCLUSIONS: TCV-116 reduced the hepatic I/R injury by inhibiting inflammatory cytokine production and by enhancing HGF production. PMID- 11889424 TI - Modulation of catecholamine responsiveness and beta-adrenergic receptor/adenylyl cyclase pathway during cardiac allograft rejection1 2. AB - BACKGROUND: This study investigated the changes of catecholamine responsiveness and beta-adrenergic receptor/adenylyl cyclase pathway during acute cardiac transplant rejection. METHODS: Isogeneic Lewis to Lewis and allogeneic Dark Agouti (DA) to Lewis rat cardiac transplants were studied 3 and 5 days after heterotopic intraabdominal transplantation (n=6/group). Myocardial blood flow (MBF), left ventricular systolic pressure (LVSP), maximum pressure development (+dP/dt), and end-diastolic pressure (LVEDP) were measured using an intraventricular balloon. Contractile response to dobutamine (5 microg/kg/min) was also assessed. In separate groups beta-adrenergic receptor density and adenylyl cyclase activity were measured in the grafts, in the recipients' native hearts and in native hearts of sham-operated controls. RESULTS: During mild to moderate rejection cardiac function indices remained unchanged, although MBF and contractile response to dobutamine decreased significantly (P<0.05) in the allogeneic group. The beta-adrenergic receptor density was significantly (P<0.05) increased in both isografts and allografts and in the native hearts of allografted recipients in comparison to native hearts of controls. Adenylyl cyclase activity showed a significant decrease (P<0.05) only in allografts. During severe rejection, LVSP and +dP/dt decreased and LVEDP increased in allografts in comparison to isografts (P<0.05). This was accompanied by a significant decrease in MBF, contractile response to dobutamine, beta-adrenergic receptor density, and adenylyl cyclase activity (P<0.05). CONCLUSIONS: Both microcirculatory disturbances and primary alteration in adenylyl cyclase activity may contribute to decreased contractile reserve in mild to moderate cardiac allograft rejection, whereas beta-adrenergic receptor density seems to be also influenced by cardiac denervation. Severe rejection leads to systolic and diastolic heart failure with complex dysregulation of the beta-adrenergic receptor/adenylyl cyclase pathway and impaired microcirculation. PMID- 11889425 TI - Epstein-Barr virus-negative precursor B cell lymphoblastic lymphoma after liver transplantation: a unique form of posttransplant lymphoproliferative disease. AB - BACKGROUND: We report a liver transplant patient with Epstein-Barr virus negative, precursor B cell lymphoblastic lymphoma diagnosed 6 months after transplantation. PATIENT: The patient was a 13-year-old boy with acute, fulminant hepatic failure of unknown etiology who underwent cadaveric liver transplantation. RESULTS: Six months after the transplant, the patient developed non-tender cervical lymphadenopathy 2 days after a reduction in prednisone dosage. The adenopathy worsened despite withdrawal of immunosuppression, and a biopsy showed precursor B cell lymphoblastic lymphoma. All investigations for Epstein-Barr virus were negative. The patient responded well to chemotherapy and is currently in complete remission 24 months after diagnosis. CONCLUSIONS: Precursor B cell lymphoblastic lymphoma has not been previously reported as a form of posttransplant lymphoproliferative disease. We discuss this unique form of posttransplant lymphoproliferative disease and briefly review the clinical and pathological spectrum of posttransplant lymphoproliferative disease. PMID- 11889426 TI - Bronchiolitis obliterans organizing pneumonia in an orthotopic liver transplant patient. AB - A 67-year-old woman was hospitalized for progressive dyspnea on exertion. She had undergone orthotopic liver transplantation (OLT) 15 months before admission. Posttransplant therapy consisted of tacrolimus, trimethoprim/sulfamethoxazole, and prednisone (the latter two were discontinued after 1 year). Physical examination revealed fine bibasilar crackles. High-resolution chest CT demonstrated bilateral, diffuse, interstitial infiltrates. Symptoms persisted on i.v. antibiotics and bronchoscopy was performed demonstrating patchy fibroplastic plugs within air spaces consistent with bronchiolitis obliterans organizing pneumonia (BOOP). Prednisone was initiated and the patient had an uneventful recovery. BOOP was initially described as an idiopathic disease process with clinical, radiographic, pathological, and prognostic features distinguishing it from bronchiolitis obliterans and idiopathic pulmonary fibrosis. BOOP has been recognized as a complication of lung and bone marrow transplantation, but the mechanism is unknown. We report a case of BOOP after OLT to highlight the risk in all transplant patients as well as the protective effect of posttransplant prednisone. PMID- 11889428 TI - Pancreas allograft biopsy: safety of percutaneous biopsy-results of a large experience. AB - BACKGROUND: Percutaneous pancreas allograft biopsy is the technique of choice for evaluation of pancreatic allograft rejection or dysfunction. The safety and success rate of the procedure with current surgical techniques is unclear. We report the complications and success rate in 426 consecutive percutaneous pancreas allograft biopsies performed at a single center. METHODS: One hundred eighty-three patients (50% simultaneous pancreas-kidney transplant, 42% pancreas after kidney transplant, 8% pancreas transplant alone) were biopsied. Biopsies were performed using an 18-gauge automated needle with ultrasound guidance. RESULTS: Thirty-seven patients had four or more biopsies (maximum, 11). Two hundred fifteen (50%) biopsies were in patients with bladder exocrine drainage, 211 (50%) in patients with enteric exocrine drainage. Of the latter, 33 (16%) were in patients with portal venous drainage. Eighty-eight percent of biopsy specimens were adequate for histologic diagnosis. There were 12 biopsy-related complications (2.8%), including eight episodes of bleeding (1.9%), five of which (1.2%) required surgical intervention and one possible extra-graft exocrine leak. Other complications included inadvertent liver (1), kidney (1), and small bowel (1) biopsy. CONCLUSIONS: Percutaneous pancreas allograft biopsy has a low rate of complication and a high success rate. PMID- 11889427 TI - The role of tacrolimus (FK506)-based immunosuppression on bone mineral density and bone turnover after cardiac transplantation: a prospective, longitudinal, randomized, double-blind trial with calcitriol. AB - BACKGROUND: Tacrolimus (FK506) is a new immunosuppressive drug in organ transplantation that has demonstrated experimentally to be more deleterious on bone mineral metabolism than cyclosporine. The purpose of this clinical study was to evaluate the effects of a tacrolimus-based immunosuppression on the skeleton and to investigate in a prospective, longitudinal, randomized, double-blind, study the effect of 0.25 microg calcitriol (1,25-dihydroxyvitamin D3) versus placebo in the prevention of bone loss and fracture rate after heart transplantion (HTx). METHODS: A total of 53 patients (5 female, 48 male, mean age: 53+/-11 years) were randomized to the study medication. Basic therapy included calcium and sex hormone replacement in hypogonadism. Bone mineral density of the lumbar spine (LS) and femoral neck (FN) were performed at baseline, after 12 and 24 months. Biochemical indexes of mineral metabolism were measured every 3 months. RESULTS: Overall bone mineral density (BMD) was significantly decreased after HTx (T-score-LS: 89+/-13%; FN: 88+/-14%). LS-BMD (% change in g/cm2) increased significantly within the study period in the calcitriol group (12 months: 7.1+/-8.1%, P<0.01; 24 months: 14.0+/-10.1%, P<0.01) and showed a positive trend in the placebo group (12 months: 4.5+/-9.3%, NS; 24 months: 6.2+/-8.0%, NS). FN-BMD in the calcitriol group was stable (12 months: 2.1+/-4.2%; NS; 24 months: -0.9+/-3.2%, NS). FN-BMD in the placebo group decreased significantly within the first 12 month follow-up period (-7.3+/-5.4; P<0.05) and stabilized within 2 years (-8.0+/-4.1%; P < 0.05). Fracture incidence was low during the study interval (first year: 5.0%, second year: 0%). Bone resorption markers decreased significantly during calcitriol therapy. CONCLUSIONS: High dose tacrolimus-based immunosuppressive regimen is associated with a rapid bone loss early after cardiac transplantation. Beyond the first 6 months after HTx, calcium, vitamin D, and hormone supplementation in hypogonadism lead sufficiently to bone mineral recovery. Besides immunosuppression, both concomitant hypogonadism and secondary hyperparathyroidism play a major role for the bone loss and should be therefore monitored and treated adequately. Low dose calcitriol should be substituted for at least 2 years as additional antiresorptive therapy. PMID- 11889429 TI - Temporal relationships between acute cellular rejection features and increased mucosal fibrosis in the early posttransplant period of human small intestinal allografts. AB - BACKGROUND: Intestinal allograft biopsies limit histopathological analysis to the superficial layers of the bowel. These biopsies allow a reasonable assessment of the histological features of acute rejection, but characteristics of chronic injury in mucosal layers remain poorly defined because of the limitations posed by endoscopic sampling. Experimental work has inferred that intestinal mucosal fibrosis may be indicative of chronic rejection; however, a temporal, graded study of mucosal fibrosis has not been performed. METHODS: A total of 79 endoscopic intestinal allograft biopsies from 12 patients obtained at 3 to 120 days posttransplantation were evaluated. Fibrosis and individual parameters of acute cellular rejection were graded according to a semiquantitative scoring system and were evaluated for potential relationships with each other. RESULTS: We found that while acute rejection tends to occur early in the posttransplant period, fibrosis of the lamina propria increases at a later time, particularly in the third and fourth month. Several individual graded parameters of acute rejection had an association with fibrosis at the same time points. CONCLUSIONS: Fibrous replacement of the lamina propria in human endoscopic allograft biopsies occurs with advancing time after transplantation. Acute rejection precedes and may have some eventual impact upon the amount of fibrosis present. A measurement of the connective tissue component of bowel transplant tissue may serve as a harbinger of long-term enteral allograft dysfunction. PMID- 11889430 TI - Zoom endoscopic evaluation of rejection in living-related small bowel transplantation. AB - BACKGROUND: This study evaluated zoom endoscopic findings according to four components in comparison with the histologic findings of acute cellular rejection (ACR) in a living-related small bowel transplantation (SBTx) patient. METHODS: A 16-year-old boy with microvillus inclusion disease underwent SBTx with a 150-cm long ileal graft from a blood-identical living-related donor. The endoscope was inserted into the distal stoma of the graft, and the mucosal architecture was observed under zoom. The observed findings were expressed by the following four components and graded as 0 to 2: H, homogeneity of mucosal change, minimal (H-0), patchy (H-1), diffuse (H-2); V, appearance of villi, stringlike (V-0), tonguelike (V-1), domelike (V-2); W, widening of crypt area, narrow (W-0), slightly widened within a width of one villus (W-1), markedly widened beyond a width of one villus (W-2); E, erythema in crypt area: no redness (E-0), sporadic erythema (E-1), diffuse erythema (E-2). RESULTS: Histologic ACR was grade 0 in 27, grade 1 in 6, grade 2 in 3, and grade 3 in 0 occasions during 11 months after SBTx. In grade 0, 1, and 2 histology, H-0/H-1/H-2 was 85.2%/14.8%/0%, 33.3%/66.7%/0%, and 0%/100%/0%, respectively, with a significant difference among the groups by grade (P<0.05). V-0/V-1/V-2 was 48.1%/51.9%/0%, 0%/100%/0%, and 0%/66.7%/33.3% (P<0.05), W-0/W-1/W-2 was 85.2%/14.8%/0%, 0%/100%/0%, and 0%/66.7%/33.3% (P<0.05), and E-0/E-1/E-2 was 100%/0%/0%, 66.7%/33.3%/0%, and 0%/100%/0% (P<0.05), respectively. CONCLUSION: Zoom endoscopic findings, graded on four components, may reflect the histologic severity of ACR and minimize the performance of biopsies in SBTx. PMID- 11889431 TI - Chronic parvovirus B19 infection in a pediatric lung transplanted patient. AB - In immunocompromised patients, clinical manifestations of human parvovirus B19 (PVB19) infection are mostly reported as acute or chronic hematological disorders. Recently, PVB19 infection has been associated with nonhematological symptoms. Four years after lung transplantation, a 9-year-old girl developed a severe anemia with reticulocytopenia requiring blood transfusion. PVB19 DNA was found by polymerase chain reaction in blood. Blood marrow aspiration revealed typical features of PVB19 infection. She was successfully treated with high dose of i.v. Ig. Then, she exhibited recurrent nonregenerative anemia requiring another course of i.v. Ig. PVB19 DNA has been persisted in blood with no specific immune response. At the same time, she suffered from several lung infection syndromes with no microorganism found except PVB19 DNA. Recurrent mild renal dysfunction was noticed with no other explanation than PVB19 infection. This report indicates that pediatric transplanted patients are at risk of chronic PVB19 infection, which can be associated with lung and/or renal disorders. PMID- 11889432 TI - Response-oriented preemptive therapy against cytomegalovirus disease with low dose ganciclovir: a prospective evaluation. AB - BACKGROUND: Preemptive therapy against cytomegalovirus (CMV) disease has succeeded in reducing the incidence of CMV disease, but the toxicity of ganciclovir remains problematic. METHODS: We prospectively evaluated the efficacy and toxicity of a preemptive protocol with ganciclovir at a reduced initial dose in 40 patients who achieved engraftment after allogeneic hematopoietic stem cell transplantation. RESULTS: Twenty-three (58%) patients had high-risk features, including transplant from an HLA-mismatched or unrelated donor, or associated acute graft-versus-host disease. CMV antigenemia assay was performed weekly, and ganciclovir was started in a risk-adapted manner, in which the initial dose of ganciclovir was fixed at 5 mg/kg/d and then adjusted based on the results of a weekly CMV antigenemia assay. In this protocol, 23 (58%) patients demonstrated positive antigenemia, and 19 (48%) received a preemptive administration of ganciclovir. Only one patient had CMV disease in the gastrointestinal system, which was successfully treated with a regular therapeutic dose of ganciclovir. Consequently, the total dose of ganciclovir was significantly less than that in a previous protocol using the conventional double dose (5 mg/kg twice daily) of ganciclovir (134 mg/kg vs. 190 mg/kg on average, P=0.046). There were no significant toxicities attributed to ganciclovir, except for neutropenia <0.5 x 109/L, which developed in three patients for 3, 4, and 14 days, respectively, with granulocyte colony-stimulating factor support. CONCLUSION: Preemptive therapy with a low initial dose of ganciclovir appeared to be effective even in high-risk patients. Further randomized controlled trial is warranted. PMID- 11889433 TI - High transforming growth factor-beta and extracellular matrix mRNA response in renal allografts during early acute rejection is associated with absence of chronic rejection. AB - BACKGROUND: A case-control study was performed to investigate whether mRNA levels of transforming growth factor-beta (TGF-beta) and various extracellular matrix molecules in renal transplant biopsy specimens, taken during acute rejection episodes within 6 months of transplantation, discriminate between patients who show deterioration of graft function and develop chronic rejection (CR+ group), and those who do not develop chronic rejection (CR- group). METHODS: Patients in both the CR+ group (n=10) and the CR- group (n=18) had at least one biopsy-proven acute rejection episode within the first 6 months after transplantation. The two groups were similar with respect to donor-, recipient-, and transplantation related clinical variables. Histologic changes (Banff classification) and the timing of the acute rejection episodes in the biopsies studied did not differ between groups. Renal cortical mRNA levels of TGF-beta1, collagen alpha1(IV), collagen alpha1(I), decorin, and the household gene glyceraldehyde-3-phosphate dehydrogenase in biopsy specimens taken during acute rejection episodes were quantified by real-time polymerase chain reaction. RESULTS: The mean TGF-beta mRNA level in the CR- group was 3.4 times higher than that in the CR+ group (P<0.04). The mean collagen IV, collagen I, and decorin mRNA levels in the CR- group were 4.2 times (P<0.05), 5.1 times (not significant), and 3.2 times (P<0.05) higher, respectively, than those in the CR+ group. The mean TGF-beta to decorin mRNA ratios between the two patient groups did not differ significantly. CONCLUSIONS: In summary, high mRNA levels for TGF-beta, collagen IV, and decorin, but not histopathologic changes, in biopsies taken during acute rejection episodes early after kidney transplantation are associated with absence of chronic rejection. We hypothesize that TGF-beta might have beneficial effects during acute rejection through its known antiinflammatory actions or as an inducer of tissue repair. PMID- 11889434 TI - Transplant tumor registry: donors with central nervous system tumors1. AB - BACKGROUND: Despite 13,000 central nervous system (CNS) tumor deaths per year in the United States, CNS tumor donors comprise only 1% of cadaveric donors recovered per year. Concern about tumor transmission may be a possible reason for this very small percentage. Both the size of the candidate waiting list and the number of deaths on the waiting list are progressively increasing because of the donor shortage. METHODS: During a 96-month period, the United Network for Organ Sharing recorded 42,340 cadaver donors of whom 397 had a past history of a CNS tumor or the cause of death listed as a CNS tumor. A total of 1,220 organs were transplanted from these 397 donors. All recipients who reported a posttransplant malignancy during a mean follow-up of 36 months were identified. RESULTS: There was no difference in patient survival of organs from CNS tumor donors when compared to donors with no CNS tumors. CNS tumor donors were not used more often for either urgent or older recipients. A total of 39 patients reported posttransplant malignancies but none of these tumors were donor-derived. There is a wide variation in the number of CNS tumor donors utilized by individual organ procurement organizations. CONCLUSIONS: The risk of tumor transmission from donors with CNS malignancies seems to be small. Certain tumors, such as glioblastoma multiforme and medulloblastoma, carry a high risk of transmission and should be avoided. The risk of tumor transmission should be weighed against the risk of the patient dying on the waiting list without a transplant. PMID- 11889435 TI - The outcome of liver grafts procured from hepatitis C-positive donors. AB - BACKGROUND: The growing prevalence of hepatitis C virus (HCV) infection in the general population has resulted in an increased frequency of potential organ donors that carry the virus. The survival of grafts from HCV+ donors has not been studied in detail. METHODS: Two study populations were examined retrospectively to assess the survival of liver grafts procured from HCV+ donors. First, we evaluated the survival of all 13 HCV+ and 103 HCV- grafts that were transplanted at our institution to HCV+ recipients from January 1, 1995 to December 31, 1999. In parallel, we analyzed a subset of the United Network for Organ Sharing (UNOS) liver transplant database from the same 5-year time period that was comprised of 14,195 adult patients for whom donor and recipient HCV serologies were known. Kaplan-Meier graft survival for both patient populations was calculated based on donor and recipient HCV serologic status. A Cox proportional hazards analysis was performed on UNOS data to identify variables independently predicting graft survival. RESULTS: For transplants performed at our institution, we found no statistically significant difference in the Kaplan-Meier graft survival of HCV+ and HCV- grafts transplanted to HCV+ recipients (P=0.68). The incidence of biopsy proven, recurrent HCV posttransplant was similar in recipients receiving either HCV+ or HCV- grafts (4/13 vs. 18/103, chi-square P=0.211). Analysis of UNOS data revealed that the survival of HCV+ grafts in HCV+ recipients was equivalent to the survival of HCV- grafts in HCV+ recipients. Unexpectedly, the survival of grafts in HCV+ recipients in general was significantly inferior to that of grafts in HCV- recipients. Multivariate analysis of all patients found recipient but not donor HCV status to be independently predictive of graft survival. CONCLUSIONS: Analysis of data from a single center and the national UNOS database suggests that transplantation of liver allografts from HCV+ donors to HCV+ recipients results in graft survival comparable to HCV- grafts transplanted to HCV+ recipients. In contrast, recipient HCV positivity is an independent predictor of graft failure compared with patients transplanted for other causes of liver disease. PMID- 11889436 TI - Diabetogenic effect of tacrolimus in South African patients undergoing kidney transplantation1. AB - BACKGROUND: Diabetes mellitus (DM) is a complication of tacrolimus therapy. This prospective study evaluated the prevalence of DM in South African black and white patients receiving tacrolimus after kidney transplantation and factors that could identify patients before transplantation who may be at risk of developing DM. METHODS: Fasting blood samples from 17 patients (11 black, 6 white) were analyzed immediately pretransplantation and at 1 and 3 months posttransplantation for glucose, HbAIC, insulin, C-peptide, free fatty acids, lipids, urea, and creatinine. Insulin resistance (IR) was calculated using the homeostasis model assessment (HOMA) and quantitative insulin sensitivity check index (QUICKI) formulas. RESULTS: Eight patients (47%) became diabetic (six black, two white), and nine patients (five black, four white) remained nondiabetic. Mean glucose concentrations in the diabetic group were significantly higher at 1 month (P=0.03) and 3 months (P=0.01). Mean insulin level was also significantly raised at 3 months (P=0.01) as was HbAIC (P=0.001). C-peptide concentrations did not change significantly in either group. Significant correlations emerged between fasting glucose concentrations at 3 months posttransplantation and initial HOMA (r=0.486; P=0.048) and initial QUICKI (r=-0.582; P=0.014). CONCLUSIONS: Occurrence of DM was high and somewhat greater in black patients. IR was the main mechanism involved, together with inadequate beta-cell compensation. IR pretransplantation predicts the subsequent development of DM. PMID- 11889438 TI - Clinical impact of human herpesvirus 6 infection after liver transplantation. AB - BACKGROUND: Reactivation of human herpesvirus 6 (HHV-6) appears to be common after transplant. Viral reactivation may result in febrile illness and may also play an immunomodulatory role that leads to indirect effects such as opportunistic infections and rejection. The objective of this study was to determine the clinical impact of HHV-6 infection after liver transplantation including both direct and indirect effects. METHODS: This was a prospective single center cohort study of 200 consecutive patients undergoing liver transplantation. Systemic serial HHV-6 viral load measurements and all clinical outcomes including development of opportunistic infections, cytomegalovirus (CMV) disease, and rejection were determined. RESULTS: HHV-6 infection (defined as viral load > or = 2 log10 copies/microg input DNA) occurred in 56/200 (28%) patients. Symptomatic disease attributable to HHV-6 alone occurred in 2/200 (1%) patients. Univariate analysis revealed HHV-6 infection was associated with the development of opportunistic infection and CMV disease. In a multivariate analysis designed to control for the level of immunosuppression, the risk of opportunistic infection increased by 3.68-fold in patients with HHV-6 infection (95% confidence interval [CI], 1.86-7.27; P=0.001). In a similar multivariate analysis, the risk of CMV disease increased by 3.59-fold in patients with HHV-6 infection (95% CI, 1.53-8.44; P=0.003). HHV-6 infection was not associated with rejection except in the subgroup of patients with rejection after 30 days posttransplant (odds ration 2.27; 95% CI, 1.09-4.77; P=0.029). CONCLUSIONS: HHV-6 reactivation after transplant is common and is associated with the development of opportunistic infections and CMV disease and possibly with a subgroup of acute rejection episodes. HHV-6 infection likely has a significant impact in transplant recipients through indirect effects of viral replication. PMID- 11889437 TI - Interleukin-1 receptor antagonist as a biomarker for bronchiolitis obliterans syndrome in lung transplant recipients. AB - BACKGROUND: The major limitation to survival after lung transplantation is bronchiolitis obliterative syndrome (BOS). BOS is a chronic inflammatory/immunologic process characterized by fibroproliferation, matrix deposition, and obliteration of the airways. The mechanism(s) that lead to fibro obliteration of allograft airways have not been fully elucidated. Interleukin-1 receptor antagonist (IL-1Ra) is a naturally occurring antagonist of the pro inflammatory cytokine IL-1 and has been associated with a number of fibroproliferative diseases. METHODS: We determined whether IL-1Ra, as compared to IL-1beta, IL-10, transforming growth factor (TGF)-beta, and tumor necrosis factor (TNF)-alpha, in the bronchoalveolar lavage fluid (BALF) from lung transplant recipients was associated with BOS. BALF was collected from three groups of patients: BOS (n=22), acute rejection (n=33), and healthy transplant recipients (n=30). RESULTS: IL-1Ra levels were significantly elevated in patients with BOS compared to healthy lung transplant recipients and patients with acute rejection (P<0.001 and P<0.05, respectively). Furthermore, when patients with BOS had their BALF analyzed from their last bronchoscopy before the development of BOS (Future BOS [FBOS] group) (n=20), their levels of IL-1Ra were also significantly elevated compared to healthy lung transplant recipients and patients with acute rejection (P<0.001 and P<0.05, respectively). Importantly, the elevated levels of IL-1Ra in the BOS and FBOS groups were not accompanied by any significant increases in IL-1beta, IL-10, TGF-beta, or TNF-alpha. CONCLUSION: These findings suggest that elevated levels of IL-1Ra may be attenuating IL-1 bioactivity during the pathogenesis of BOS and creating a local environment that favors fibroproliferation and matrix deposition. PMID- 11889439 TI - Combined liver and kidney transplantation in a patient with sickle cell disease. AB - Sickle cell intrahepatic cholestasis is a potentially fatal end-organ complication of sickle cell anemia. Renal involvement in sickle cell anemia is common, and in some cases, can present as acute renal failure. Although renal transplants have been performed in patients with sickle cell anemia since the late 1960s and a number of liver transplants have been recently performed for these complications, there has not been experience with dual organ transplantation for sickle cell anemia-related complications. We describe the case of a patient with sickle cell anemia who underwent successful combined liver and kidney transplantation after the development of acute sickle cell intrahepatic cholestasis and renal failure requiring continuous venovenous hemodialysis. The patient underwent a successful combined liver and kidney transplant with limited perioperative complications and preserved allograft function. At 22 months posttransplant, the patient expired as a result of an acute pulmonary embolus in the setting of bilateral hip fractures. Autopsy revealed no evidence of liver or kidney allograft rejection and evidence of chronic sickle cell nephropathy in the native kidney. Combined liver and kidney transplantation is a viable therapeutic option in patients with severe end-organ effects of sickle cell anemia. PMID- 11889440 TI - Disseminated varicella infection in adult renal allograft recipients: four cases and a review of the literature. AB - Disseminated varicella-zoster (VZV) infection is a rare complication after renal allotransplantation in adults. We report four patients, among them one with combined VZV and cytomegalovirus infection. The main complications were hepatitis, pneumonitis, and disseminated intravascular coagulation. A review of the literature from 1981 to 2000 revealed 34 additional cases of disseminated varicella infection in adult renal allograft recipients with an overall mortality of 34%. Among these patients 82% suffered from primary varicella, 18% had a reactivation. High-dose acyclovir therapy combined with reduction of immunosuppression lead to reduction of mortality from 53% before 1990 to 22% after 1990. No immunosuppressive drug is significantly associated with a higher risk of disseminated VZV infection. Immunization against VZV in adults is still a matter of controversy. Whereas passive immunization is performed only for prophylactic but not therapeutic purpose, active immunization is routinely performed in children and may also be recommended for adults before renal transplantation. PMID- 11889441 TI - Effect of kidney transplantation on bone mass and body composition in males. AB - BACKGROUND: Bone loss is a frequent and well-known complication in the first months after renal transplantation, but there are no data considering body composition variables (bone, fat, and lean mass) together in transplant recipients. This prospective study investigated total body bone density, fat mass, and lean mass before and 1, 2, 3, 4, and 6 months after renal transplantation in male patients who underwent hemodialysis. METHODS: Twenty consecutive renal transplant male patients aged 23-64 years (mean, 40 years; median, 41 years) received one of two immunosuppressive therapies (cyclosporine+methylprednisolone, or cyclosporine+methylprednisolone+azathioprine). The bone, fat, and lean mass of the total body and its related subregions were assessed by means of dual X-ray photon absorptiometry. Mixed factorial analysis of variance for repeated measurements was used for the statistical analysis. RESULTS: During the 6 months after transplantation, there was a reduction in trabecular bone mass in the spine, ribs, and pelvis total body subregions; the reduction was statistically significant in the last two subregions. There was no statistically significant difference in the lean mass of the total body or its subregions over time, but there was a statistically significant increase in the fat mass of the total body and all of its subregions; the increase in total and trunk fat mass seemed to be greater in the patients not receiving azathioprine. CONCLUSIONS: Up to 6 months after renal transplantation in male patients who underwent hemodialysis, there is a marked increase in fat mass, a significant loss of trabecular bone mass, and no change in cortical bone and lean mass. PMID- 11889442 TI - Cost-effectiveness of cadaveric and living-donor liver transplantation. AB - BACKGROUND: Cadaveric liver transplantation (5-year survival >80%) represents the standard of care for end-stage liver disease (ESLD). Because the demand for cadaveric organs exceeds their availability, living-donor liver transplantation has gained increasing acceptance. Our aim was to assess the marginal cost effectiveness of cadaveric and living-donor orthotopic liver transplantation (OLT) in adults with ESLD. METHODS: Using a Markov model, outcomes and costs of ESLD treated (1) conservatively, (2) with cadaveric OLT alone, and (3) with cadaveric OLT or living-donor OLT were computed. The model was validated with published data. The case-based scenario consisted of data on all 15 ESLD patients currently on our waiting list (3 women, 12 men; median age, 48 years [range, 33 59 years]) and on the outcome of all OLT performed for ESLD at our institution since 1995 (n=51; actuarial 5-year survival 93%). Living-donor OLT was allowed in 15% during the first year of listing; fulminant hepatic failure and hepatocellular carcinoma were excluded. RESULTS: Cadaveric OLT gained on average 6.2 quality-adjusted life-years (QALYs) per patient compared with conservative treatment, living-donor OLT, an additional 1.3 QALYs compared with cadaveric OLT alone. Marginal cost-effectiveness of a program with cadaveric OLT alone and a program with cadaveric and living-donor OLT combined were similar (E 22,451 and E 23,530 per QALY gained). Results were sensitive to recipient age and postoperative survival rate. CONCLUSIONS: Offering living-donor OLT in addition to cadaveric OLT improves survival at costs comparable to accepted therapies in medicine. Cadaveric OLT and living-donor OLT are cost-effective. PMID- 11889443 TI - Hypertension is an independent predictor of delayed graft function and worse renal function only in kidneys with chronic pathological lesions. AB - BACKGROUND: Delayed graft function (DGF) has been identified as one of the principal correlates of poor graft survival in cadaveric renal transplantation. However, its risk factors and clinical predictors have been poorly elucidated. METHODS: We analyzed the risk factors of DGF with a specific emphasis on the role of histological damage of donor kidney. Then, we also studied the impact of DGF, and donor factors affecting DGF, on kidney graft function over the first year after engraftment in 100 consecutive cadaveric renal transplant (Tx) recipients. RESULTS: The organs displaying DGF (n=48) had a significantly higher degree of glomerular sclerosis and tubular atrophy (P<0.01), as well as of interstitial fibrosis and vascular damage (P<0.02) in time-zero biopsies. In patients who received an "ideal" organ for Tx (total histological score < or = 4), DGF showed a strong relationship with Deltaage D-R (70% increase of risk for donors 10 years older than recipients), and with the histological score (odds ratio 1.34). In contrast, donor hypertension was the most relevant variable independently associated with DGF (odds ratio 19.4) in patients receiving a suboptimal organ (histological score >4). Moreover, DGF and donor hypertension adversely affected graft function at 1 year, but only in Tx patients with a histological score >4 in time-zero biopsy. Of note, both patients with and those without DGF showed a very low incidence of biopsy-proven acute rejection (8.5 and 6.8%, respectively) and a rather short cold ischemia time (<16 hr). CONCLUSION: Our findings suggest that the quality of the transplanted organ and the occurrence of DGF are strictly related to each other and can influence graft function through apparently nonimmune mechanisms. In addition, long-standing donor hypertension is a strong independent variable affecting both DGF and graft function of suboptimal cadaveric kidneys, at least up to 1 year. PMID- 11889444 TI - Temporary auxiliary liver transplantation from a living donor to an adult recipient with familial amyloid polyneuropathy. AB - A 33-year-old patient with familial amyloid polyneuropathy (FAP) underwent temporary auxiliary partial orthotopic liver transplantation (APOLT) from a living donor with a small-for-size graft. The auxiliary left lobar graft, which weighed only 230 g, was orthotopically transplanted after resection of the recipient's left lobe. The right portal vein was transected to induce compensatory hypertrophy of the left lobar graft. Posttransplant computed tomography showed atrophy of the native liver and hypertrophy of the graft, the volume of which had increased to 446 ml by postoperative day 41. The remnant native liver was removed 6 weeks after APOLT, and there were no signs of liver dysfunction during the postoperative course. Our experience with this case suggests that temporary APOLT is the treatment of choice, guaranteeing a sufficient margin of safety for both donor and recipient, in living donor liver transplants for FAP where the donor's left lobe is disproportionately small. PMID- 11889445 TI - Diameter of the infrarenal aorta and the iliac arteries in children: ultrasound measurements. AB - BACKGROUND: In adults, the diameters of the infrarenal abdominal aorta and the iliac vessels on ultrasound (US) are known. Similar values have not yet been reported in children. The purpose of this study was to establish a nomogram for these diameters in children. Therefore, the aim is to delineate another factor to limit intraoperative graft lost. METHODS: Studies were performed in 176 healthy children from 1 to 16 years of age. The diameters of the aorta and iliac arteries were measured by B-mode ultrasound at predefined sites. A correlation of the vessel diameter, age, gender, weight, height, body mass index, and body surface area (BSA) was performed. RESULTS: At all measured points, vessel diameters were significantly (P<0.0001) larger in boys than in girls. There was a significant (P<0.0001) increase of all the vessel diameters over age in both sexes. Vessel diameters correlate positively with age, gender, weight, height, and BSA. The highest correlation was found to be with BSA (r > or = 0.8, P<0.0001). Nomograms for each arterial diameter could be established for males and females separately. CONCLUSION: Normal US values of the diameter of the infrarenal aorta and the iliac vessels have been determined for children. The change in diameter strongly correlates with BSA. The nomograms can be of great help in the pretransplantation assessment of these vessels. PMID- 11889446 TI - Inhibition of CD40-mediated endothelial cell activation with antisense oligonucleotides. AB - BACKGROUND: CD40-CD154 interactions play a pivotal role in the amplification of immune responses and, as such, represent an attractive target for immune intervention in a number of disease indications. We have previously shown that binding of human CD154 expressed on the Jurkat D1.1 cell line to porcine CD40 on pig aortic endothelial cells (PAECs) can lead to up-regulation of vascular cell adhesion molecule (VCAM)-1 and MHC class II. This activation can be completely inhibited by the addition of a monoclonal antibody (mAb) to human CD154. In this study, we explore an alternative approach to blocking this pathway with antisense oligonucleotides (ASOs). METHODS: Ten ASOs were generated on the basis of the porcine CD40 cDNA sequence. The ASOs that were found to reduce CD40 expression on PAECs were analyzed for their ability to reduce CD40-mediated PAEC activation. RESULTS: Four ASOs were found to significantly lower surface expression of porcine CD40 on PAECs 48 hr after transfection. Eight of the ASOs were seen to lead to mRNA cleavage products by ribonuclease protection assay. Of the four ASOs tested in the PAEC activation assay, one (ASO-9) showed a dramatic inhibition of PAEC activation (IC50 approximately 1 nM) results comparable to the use of a blocking mAb. Furthermore, we compared the effect of CD40 ASO on tumor necrosis factor alpha receptor signaling, in which we observed no effect, which confirmed ASO specificity. CONCLUSIONS: These results indicate that a CD40-dependent activation pathway can be inhibited with an ASO with high potency and specificity. ASO could be an attractive alternative therapy to the use of mAbs. PMID- 11889447 TI - Is there MHC Class II restriction of the response to MHC Class I in transplant patients? AB - BACKGROUND: In this study, we evaluated distinct HLA-DRB1 alleles to determine class II restriction of the production of HLA-A2-specific antibodies in renal transplant patients. METHODS: Data from 217 renal transplant patients who received an HLA-A2-mismatched renal graft were analyzed with regard to HLA-A2 humoral responsiveness. High-resolution DNA typing of class II HLA-DR alleles was performed by polymerase chain reaction-sequence-specific primer. Patients who had one of the following eight HLA-DRB1 alleles were included in the study: -*0101, *0301, -*0401, -*0701, -*1101, -*1301, -*1401, and -*1501. Serum samples were screened posttransplantation with the standard complement-dependent cytotoxicity procedure. In addition, recombinant HLA-A2 monomers (the "MonoLISA" assay) were used as a target for the detection of HLA-A2 group-specific antibodies. The following HLA-A2 amino acid positions (termed "epitopes") that are responsible for the induction of an antibody response were defined: 74H, 65-66GK, 62G, 114H, 142-145TTKH, and 107W-127K. The definition of the "HLA-DR permittors" of anti-HLA A2 response was based on a "class II restriction table" designed for this purpose. Prediction of immunogenic and/or nonimmunogenic HLA-A2 peptides was based on an MHC database. RESULTS: The HLA-DRB1-*0101 and -*1401 alleles had a trend toward a positive correlation with the production of HLA class I-specific antibodies against the HLA-A2 shared (public) epitopes 65-66GK and -62G, respectively. Only the DRB1-*1501 allele had higher trend toward a positive correlation with the production of antibodies against the HLA-A2 private (74H) epitope. In 42 patients with the HLA-DRB1-*1501 allele, 11 (26%) patients produced HLA-specific antibodies against the HLA-A2 group of epitope(s). Moreover, in these patients, spreading of the alloreactivity against "other" HLA antigens was detected. Many of these other HLA antigens did not belong to HLA-A2 group but had newly defined shared epitopes with this group. Furthermore, the epitope prediction, based on an MHC database, revealed differences in the ligation strength (score) to the HLA allele (class I and II) for a specific HLA A2 peptide in the 42 patients (responders and nonresponders). CONCLUSIONS: The data presented in this paper suggest that the HLA class II allele and the type of the bound allopeptide may influence the humoral and cellular response. The immunogenicity of these allopeptides could be predicted with an MHC database (high-scored peptide=activating peptide and low-scored peptide=suppressor peptide). In the future, production of synthetic peptide analogues, on the basis of these predictions, could be used for induction of T-cell anergy and/or tolerance. In the short term, algorithms, on the basis of our approach, could be tested for influence on graft survival and allosensitization in current high quality data sets. PMID- 11889448 TI - Heart allograft tolerance without development of posttransplant cardiac allograft vasculopathy in chimerism-based, drug-induced tolerance. AB - BACKGROUND: Recently, we have described a drug (cyclophosphamide [CP] plus busulfan [BU])-induced skin allograft tolerance in mice that can regularly overcome fully H-2-mismatched barriers. Using this method, we have investigated whether or not this regimen can prolong the survival of heart allografts and inhibit the development of posttransplant cardiac allograft vasculopathy (CAV). METHODS: The components of the method are intravenous administration of 1 x 108 allogeneic spleen cells on day 0, intraperitoneal injection of 200 mg/kg of CP and 30 mg/kg of BU on day 2, and intravenous injection of T cell-depleted 1 x 107 allogeneic bone marrow cells from the same strain of mice on day 3. Heart grafting was performed on day 28. Chimerism in peripheral blood was followed by flow cytometric analysis, and histological analysis was performed at various times after grafting. RESULTS: In a fully major histocompatability complex (MHC) mismatched combination of B10.D2 (H-2d, IE+)-->B10 (H-2b, IE-), stable, multilineage-mixed chimerism was observed permanently. B10.D2 heart grafts were accepted permanently in a donor-specific manner, and posttransplant CAV did not develop. CONCLUSIONS: These results demonstrated that the drug-induced tolerance recently established by us can regularly induce a long-lasting heart allograft tolerance without development of CAV. PMID- 11889449 TI - Expression of growth arrest-specific gene 6 and its receptors in a rat model of chronic renal transplant rejection. AB - BACKGROUND: Growth arrest-specific gene 6 (Gas6) is involved in a number of cell functions that include proliferation of vascular smooth muscle cells and mesangial cells. The proliferation of these cells is a feature of chronic rejection (CR) after kidney transplantation. Therefore, we examined the gene expression of Gas6 and its receptors Rse, Axl, and Mer in a rat model of CR. METHODS: The rat model of CR was established in Lewis rat recipients of Fisher kidney transplants. The level of mRNA was measured by real-time quantitative reverse transcription-polymerase chain reaction. The proteins were detected by immunohistochemical staining and Western blot analysis. RESULTS: Gas6 mRNA was extensively expressed in kidney tissue of both allografts and isografts. There was significant increase in expression of Gas6 mRNA in allografts at 4 weeks posttransplantation. Immunohistochemical study showed that Gas6 and its receptor Rse proteins were highly expressed in kidney tissue. Western blot analysis has also confirmed that Gas6 and Rse proteins are expressed in kidney tissue. CONCLUSIONS: These findings suggest that Gas6 and its receptors have an as yet undefined role in kidney function and/or development and may be involved in the pathogenesis of CR. The action of Gas6 in rat kidney is mainly mediated through the Rse receptors rather than the Axl and Mer receptors. PMID- 11889450 TI - Topical use of cidofovir induced acute renal failure. AB - BACKGROUND: Cidofovir has antiviral activity against a wide spectrum of DNA viruses. Several small studies have focused on the efficacy of topical cidofovir in various viral-induced diseases. We report a systemic complication of such therapy. CASE REPORT: A bone marrow transplant recipient with chronic renal failure developed genital condylomas resistant to standard therapy. After topical cidofovir application (1% once daily for 5 days, then 4% for 12 days), the lesions improved while local erosions appeared. Acute renal failure with features of tubular acidosis occurred at day 19. Spontaneous recovery was observed after cidofovir withdrawal. CONCLUSION: We describe for the first time acute renal failure after topical cidofovir in an immunosuppressed patient with prior renal insufficiency. This method of administration should be avoided on abraded skin and should be carefully monitored. PMID- 11889451 TI - Treatment of hyperhomocysteinemia with folic acid reduces oxidative stress in renal transplant recipients. AB - BACKGROUND: We conducted a prospective, uncontrolled, open study to assess the relationship between homocysteine (tHcy) and oxidative stress in chronic, stable, renal transplant recipients (RTR). METHODS: Included in the study were 17 chronic, stable RTR. All the patients received folic acid (5 mg/day). tHcy and total antioxidant capacity (TAOC) were measured before and at the end of the study period. RESULTS: Mean tHcy concentration was 26+/-10 micromol/L. tHcy significantly decreased during the study period (26+/-10 vs. 18+/-7 micromol/L; P<0.001). There was a significant inverse relationship between TAOC and tHcy (r= 0.33; P=0.01). TAOC significantly increased during the study period (1.49+/-0.23 1.78+/-0.6; P<0.001). There was an inverse relationship between the variation in tHcy and the variation in TAOC (r= -0.44; P=0.01). CONCLUSION: Our results demonstrate that hyperhomocysteinemia contributed to increased oxidative stress in RTR. tHcy-lowering treatment with folic acid may lower oxidative stress. PMID- 11889452 TI - Recombinant human tumor necrosis factor receptor fusion protein as complementary treatment for chronic graft-versus-host disease. AB - BACKGROUND: Chronic graft-versus-host disease (cGVHD) remains one of the major late complications in allogeneic bone marrow transplantation (BMT). Prolonged immunosuppression often results in significant morbidity and mortality. Cytokine dysregulation is implicated in the pathophysiology of cGVHD, and tumor necrosis factor-alpha (TNF-alpha) plays a central role. METHODS: Recombinant soluble TNF receptor (Enbrel) was explored for the use of steroid-dependent cGVHD in 10 patients. Enbrel was given as a subcutaneous injection twice weekly for 4 weeks followed by once weekly for 4 more weeks. Progression or regression of cGVHD was monitored closely by regular clinical follow-up. RESULTS: Eight patients finished the 8-week treatment course without adverse side effect. Seven of them showed improvement (subjectively and/or objectively) in cGVHD. Steroid taper was initiated as early as 1 month. CONCLUSIONS: This preliminary encouraging result merits additional studies to optimize Enbrel as a potential complementary therapy for resolution of steroid-dependent cGVHD. PMID- 11889453 TI - Long term outcome of lung transplant is predicted by the number of HLA-DR mismatches. PMID- 11889454 TI - Intestinal lymphoma perforations as a consequence of highly effective anti-CD20 antibody therapy. PMID- 11889455 TI - Distinct regions of loss of heterozygosity on 22q in different sites of head and neck squamous cell carcinomas. AB - BACKGROUND: Frequent loss of heterozygosity (LOH) has been reported in many types of cancer, including head and neck carcinomas. Somatic deletions involving specific chromosomal regions are strongly associated with inactivation of the allele of a tumor suppressor gene located within the deleted region. In most studies concerning LOH in head and neck squamous cell carcinomas (HNSCC) the different anatomical sites are not distinguished. The behavior of tumors arising at various sites differs significantly, however, suggesting different intrinsic tumor properties. In this study we compared the LOH on 22q and its relationship to clinicopathological parameters at the three major sites of HNSCC: oral cavity, larynx and pharynx. MATERIAL/METHODS: LOH and microsatellite instability (MSI) were studied using seven polymorphic microsatellite markers mapped to the 22q11 q13.3 region in 37 oral, 32 laryngeal, and 31 pharyngeal carcinomas. RESULTS: Two separate regions of LOH were identified in the laryngeal (22q11.2-12.1) and oral cavity (22q13.1-13.31) tumors. When the different anatomical sites were compared, a statistically significant difference was found between the presence of LOH at D22S421 (p<0.001), D22S315 (p=0.014) and D22S929 (p=0.026) in the laryngeal tumors. CONCLUSIONS: These data suggest that distinct regions on 22q are involved in LOH in oral cavity and laryngeal tumorigenesis, but do not support a similar association between the development of pharyngeal tumors and genes located on 22q. These findings implicate the presence of different tumor suppressor genes mapping to distinct regions on chromosome 22q in oral and laryngeal carcinomas. PMID- 11889456 TI - Loss of heterozygosity in primary lung cancer using laser capture microdissection and WAVE DNA fragment analysis techniques. AB - BACKGROUND: A number of molecular changes observed by varied conventional methods, including loss of heterozygosity (LOH) on chromosome 3, have been associated with primary lung cancer. To further define the locus of chromosome 3p allele loss in lung cancer, we performed LOH study by using innovative laser capture microdissection and WAVE DNA Fragment Analysis. MATERIAL/METHODS: Thirty eight paired specimens from patients with adenocarcinoma of the lung were used for this study. Formalin-fixed, paraffin-embedded tissue from normal stromal cells or lymphocytes and adenocarcinoma were collected using laser capture microdissection. DNA was extracted and amplified by PCR using six polymorphic DNA markers for chromosome 3. PCR products were analyzed by both gel electrophoresis and WAVE DNA Fragment Analysis. RESULTS: LOH at 3p22-24 was found in tumor cells from twelve out of thirty-eight patients (32%) when analyzed by WAVE DNA Fragment Analysis and LOH was found in tumor cells from nine out of thirty-eight patients (23%) when analyzed by gel electrophoresis. LOH was found in normal control from one out of thirty-eight patients. CONCLUSIONS: 1. Our results suggest putative tumor suppressor gene(s) is present in a region at 3p22-24, which may play a role in carcinogenesis of lung cancer. 2. Laser capture microdissection is essential tool for defined LOH studies. 3. WAVE DNA Fragment Analysis is an accurate, sensitive and automated tool for analysis of DNA fragments. PMID- 11889457 TI - Resistance of Klebsiella pneumoniae strains producing and not producing ESBL (extended-spectrum beta-lactamase) type enzymes to selected non-beta-lactam antibiotics. AB - BACKGROUND: Bacteria of the Klebsiella genus may cause numerous infections in human, which are often treated with beta-lactam antibiotics. The fundamental mechanism of Klebsiella resistance to penicillins or cephalosporins involves the production of enzymes--extended-spectrum beta-lactamases (ESBLs). Because of resistance of many Klebsiella spp. strains to beta-lactams, alternative antibiotic therapy can make use of aminoglycosides and quinolones. MATERIAL/METHODS: The study analyzed the prevalence of ESBL -type enzymes among 256 Klebsiella pneumoniae strains isolated from various clinical materials collected from patients hospitalized between 1997 and 2000. ESBLs were detected by double-disk synergy test (DDST). The prevalence of strains resistant to selected aminoglycosides (gentamicin, amikacin, netilmicin) and quinolones (ciprofloxacin, norfloxacin, nalidixic acid) in the particular years was analyzed. Drug sensitivity was determined by disk-diffusion method according to the recommendations of the National Reference Center for Microbial Drug Sensitivity. RESULTS: During the analyzed time interval, a significant increase of the number of K. pneumoniae ESBL(+) strains was noted: in 1997 - 16.5% (14/85) and in 2000 - 40.4% (22/54) (p<0.001). Among the ESBL(+) strains, an increase of the number of strains resistant to the tested antibiotics, except for nalidixic acid, was demonstrated CONCLUSIONS: A statistically significant increase of multidrug-resistant K. pneumoniae strains, including strains producing ESBLs, was demonstrated in the analyzed material. PMID- 11889458 TI - Leptin and estradiol as related to change in pubertal status and body weight. AB - BACKGROUND: Previous studies have examined the relationship between leptin, body mass, and pubertal status. The present study directly compares the relationship of leptin and estradiol in 5 groups of girls with different combinations of pubertal status and weight. MATERIAL/METHODS: We studied girls with idiopathic precocious puberty, age-matched non-obese prepubertal girls, age-matched obese prepubertal girls, normal pubertal girls, and obese pubertal girls (n=12/group). RESULTS: Leptin levels were significantly higher in obese pubertal girls (26.6+/ 8.4 ng/mL) than in all others. Leptin levels were significantly higher in obese prepubertal girls (20.7+/-10.9 ng/mL) (mean+/-SD) compared to girls with precocious puberty (7.7+/-5.4 ng/mL, p<0.004), non-obese prepubertal girls (5.55+/-3.6 ng/mL, p<0.001), and normal pubertal girls (4.8+/-2.9 ng/mL, p<0.001). Leptin level did not correlate significantly with estradiol level. Leptin-SDS (standard deviation score), which corrects leptin level for gender, pubertal stage, and BMI, was significantly lower in the obese pubertal group than in the 3 non-obese groups. CONCLUSIONS: These findings confirm previous studies that body mass, chronological age and pubertal stage are determinants of leptin level. However, these are not the only factors determining leptin level as evidenced by persistent differences in leptin level between obese and non-obese pubertal girls even when correcting for pubertal stage and BMI. While the present study provides no new answers to the question of pubertal regulation, it provides a direct comparison of combinations of weight, pubertal stage, and leptin level, as background for future studies. PMID- 11889459 TI - Evaluation of oxidative stress indices during treatment in pregnant women with intrauterine growth retardation. AB - BACKGROUND: We investigated treatment-related changes in oxidative stress indices in intrauterine growth retardation (IUGR) during pregnancy. Four parameters of cell membrane destruction were measured: malondialdehyde, Schiff bases, lipid peroxides, and conjugated dienes. MATERIAL/METHODS: The study population included pregnant women under treatment in the Department of Obstetrics and Gynecology at the Medical University of Lodz in 1997-1999, divided into two groups: (I) - 30 healthy pregnant women, (II) - 31 pregnant women with a clinical and ultrasound diagnosis of IUGR. The course of treatment consisted of ten days of intravenous injections of cocarboxylase, Vitamin C, and solcoseryl as antioxidant factors. The parameters of oxidative stress were measured on the first day of hospitalization, after five days of treatment, and on the 10th day of treatment. RESULTS: The mean value of MDA concentration in Group I was 2.03, vs. 2.39 in Group II. The mean value of conjugated dienes was 2.40 in Group I and 2.41 in Group II. The Schiff bases concentration in Group I was 8.03, as opposed to 8.83 in Group II. The concentration of lipid peroxides was 0.113 in Group I and 0.134 in Group II. We found that all the parameters of oxidative stress were higher in IUGR than in normal pregnancy. At the end of treatment a decrease was observed in all oxidative stress parameters. CONCLUSIONS: IUGR is correlated with an increase in membrane damage parameters. Therapy for IUGR decreased the values of all indices of oxidative stress. PMID- 11889460 TI - Determination of lactic acid level in systemic liquids in children with progressive encephalopathies. AB - BACKGROUND: This article reports the results of research into the activities of lactic acid concentrations in the body fluids of children with progressive encephalopathies (PE) in comparison to patients with non-progressive encephalopathies (NPE) and those with non-progressive encephalopathies with concomitant epilepsy (NPEE). The study was designed to determine whether there is difference between the serum and CSF lactic acid concentrations in children with progressive encephalopathies (PE), static (non-progressive) encephalopathies (NPE) and non progressive encephalopathies with concomitant epilepsy (NPEE), and whether the clinical status correlates with the concentration of these biochemical markers in children with PE. MATERIAL/METHODS: The assessment involved 138 children of both sexes, whose age ranged between 8 months and 15 years, diagnosed and treated in the Neurology Department at the Pediatric Clinic of the Silesian Medical Academy in Katowice between 1995 and 1997. Lactate concentrations were determined in serum and cerebro-spinal fluid and analyzed statistically. RESULTS: The findings showed higher serum and CSF concentrations in children with PE than in patients who manifested non-progressive forms of encephalopathy. The degree of clinical symptom aggravation in PE children was likewise analyzed and compared to the values of lactate concentrations in body fluids; however, no correlation was found between these parameters. CONCLUSIONS: Children with progressive encephalopathies present higher lactate concentrations in serum and cerebrospinal fluid than patients with static (non-progressive) encephalopathy. PMID- 11889461 TI - Eradication of Helicobacter pylori in duodenal ulcer disease tetracycline & furazolidone vs. metronidazole & amoxicillin in omeprazole based triple therapy. AB - BACKGROUND: The object of the study was to study the efficacy and safety of furazolidone and tetracycline compared to metronidazole and amoxicillin in an omeprazole based triple therapy in a prospective randomized-blind-clinical trial. MATERIAL/METHODS: Patients with endoscopically verified active duodenal ulcer disease in the presence of Helicobacter pylori infection were eligible to enter the study. Endoscopy was performed a day before and 6-8 weeks after the cessation of treatment. H. pylori status was assessed by histologic examination (Giemsa stain) of biopsy specimens were taken from the antrum and corpus. H. pylori eradication was defined as absence in histology of the biopsy specimens at the second endoscopy. Ulcer healing was considered as decrease in ulcer size to less than 20% of its primary size. Patients were randomly assigned to receive omeprazole 20 mg, amoxicillin 1000 mg and metronidazole 500 mg (OAM group) or omeprazole 20 mg, tetracycline 500 mg and furazolidone 200 mg (OTF group). All medications were taken twice daily, for 2 weeks. RESULTS: Out of 111 patients enrolled in the study, 108 completed a course of treatment and underwent a follow up endoscopy, with 54 patients in each group. H. pylori eradication was achieved in 52 patients (96.3% - 95% CI: 91.27-100) in OTF group and 45 patients (83.3% - 95% CI: 73.35-93.25) in OAM group (P=0.015). Our study showed the superiority of OTF vs. OAM regimen with a 13% increment in eradication rate, with only occasional severe side effect. CONCLUSIONS: In conclusion OTF regimen is a safe, cheaper and effective alternative for OTF regimen and we recommend it to be used especially in developing countries. PMID- 11889462 TI - Cdk4 and p27Kip1 play a role in PLC-gamma1-mediated mitogenic signaling pathway of 18 kDa FGF-2 in corneal endothelial cells. AB - PURPOSE: To determine whether PLC-gamma1 enzyme activity is essential for cell proliferation in response to FGF-2 stimulation and to investigate the effect of PLC-gamma1 activation on cell division and on processes that regulate cell cycle progression. METHODS: Cell proliferation was assayed using a colorimetric method to determine the number of viable cells. Subcellular localization of proteins was determined by immunocytochemical analysis, and expression of the proteins was analyzed by immunoblotting. PLC activity was determined by measuring the total inositol phosphates. RESULTS: When CEC were treated with FGF-2, a prolonged and continuous FGF-2 exposure was required for both PLC enzyme activation and cell proliferation. However, there was a long lag period between the PLC enzyme activation and cell proliferation: the maximum enzyme activity was reached 8 h after FGF-2 stimulation, but no cell proliferation was observed in the cells exposed to FGF-2 for 8 h. Using neutralizing anti-PLC-gamma1, PLC-beta1, or PLC delta1 antibodies, we further demonstrated that PLC-gamma1 accounts for the hydrolysis of total phosphoinositides (PI) and cell proliferation mediated by FGF 2. The role of PLC-gamma1 linking to the cell cycle was then determined by the blockades of FGF-2 action on Cdk4 and p27-Kip1. Interestingly, FGF-2 both upregulates Cdk4 synthesis and facilitates the nuclear import of the molecule from the cytoplasm, whereas it facilitates the nuclear export of p27-Kip1 to the cytoplasm without affecting synthesis of the molecule. The neutralizing anti-PLC gamma1 antibody completely abolishes the FGF-2 activity on Cdk4, both at the synthetic level and at the level of translocation, and the PLC-gamma1 antibody blocks the nuclear export of p27-Kip1. CONCLUSIONS: These data indicate that PLC gamma1 ultimately leads to activation of the cell cycle machinery to induce cell proliferation mediated by FGF-2. It does so by upregulating Cdk4 expression and by facilitating the nuclear import of the molecule and nuclear export of the Cdk inhibitor (p27-Kip1) to the proteolysis site, the cytoplasm. PMID- 11889463 TI - A goniolens for clinical monitoring of the mouse iridocorneal angle and optic nerve. AB - PURPOSE: Due to the increasing importance of mouse models and mouse genetics in ophthalmic research, we have developed a goniolens for mice. METHODS: The size and basic design parameters of a human goniolens were adapted to the mouse eye. RESULTS: The goniolens is straightforward to use and allows non-invasive visualization of the structures of the anterior chamber angle, including Schlemm's canal, trabecular meshwork, iris and anterior surface of the peripheral ciliary body. Goniophotography using the mouse lens allows documentation of anterior chamber angle abnormalities, and facilitates comparisons of changes to these structures in the same eye over time. In addition, high quality magnified views of the optic nerve, retinal vessels and posterior retina are easily obtained. CONCLUSIONS: This goniolens is an important new tool for studying the genetic and clinical etiology of anterior segment dysgenesis and glaucoma in mice. PMID- 11889464 TI - Gene transfer of dominant-negative RhoA increases outflow facility in perfused human anterior segment cultures. AB - PURPOSE: To investigate the regulation of expression and the role of the RhoA gene in the human trabecular meshwork (TM). To attempt to modulate outflow facility by gene transfer of the RhoA gene's dominant-negative mutant protein. METHODS: Total RNA extracted from cultured human trabecular meshwork (HTM) cells treated with outflow facility drugs were analyzed by northern blot hybridization using an amplified human RhoA cDNA from plasmid pZip-RhoA wild type (wt) [1]. A dominant-negative form of RhoA (single amino acid substitution of Thr19 to Asn) was placed under the control of the CMV promoter and inserted into a replication deficient adenoviral vector by overlapping recombination (AdhRhoA2). AdhRhoA2 was infected into perfused anterior segment cultures from post-mortem human donors and HTM and Schlemm's canal cells in culture. Changes in outflow facility (flow/pressure) were calculated as percent changes from baseline values (C0), pooled into treated and control groups and expressed as the mean plus minus standard error. HTM and Schlemm's Canal (SC) cells were fluorescently double labeled for the RhoA protein and actin, paxillin, or ZO-1. RESULTS: Transcription of RhoA in HTM cells was not considerably affected by treatment of the cells with cytoskeletal/outflow facility drugs. At 66 h post-injection, anterior segments treated with AdhRhoA2 (n=9) exhibited an increase in outflow facility of 32.5 +/- 7.7% while that of the vehicle-injected controls (n=6) was 5.1 +/- 4.0% (p=0.02). HTM cells treated with AdhRhoA2 showed a marked change in morphology with a reduction in actin stress fibers and of the focal adhesion-containing protein, paxillin. Confluent monolayers of SC cells infected with AdhRhoA2 were devoid of peripheral ZO-1 staining indicating a loss of intercellular junctions. CONCLUSIONS: In the HTM cells, cytoskeletal/outflow facility drugs do not seem to affect the levels of RhoA mRNA, possibly suggesting the importance of mRNA availability to allow rapid turnover of its function. Gene transfer of inactive RhoA to the intact human TM results in an increase in outflow facility. This increase appears to be correlated with a loosening of the cell-substrate and cell cell attachments in the cells of the outflow pathway. Adenoviral vectors carrying the dominant negative form of RhoA could potentially be utilized as a gene therapy to modulate outflow facility. PMID- 11889465 TI - FACL4, encoding fatty acid-CoA ligase 4, is mutated in nonspecific X-linked mental retardation. AB - X-linked mental retardation (XLMR) is an inherited condition that causes failure to develop cognitive abilities, owing to mutations in a gene on the X chromosome. The latest XLMR update lists up to 136 conditions leading to 'syndromic', or 'specific', mental retardation (MRXS) and 66 entries leading to 'nonspecific' mental retardation (MRX). For 9 of the 66 MRX entries, the causative gene has been identified. Our recent discovery of the contiguous gene deletion syndrome ATS-MR (previously known as Alport syndrome, mental retardation, midface hypoplasia, elliptocytosis, OMIM #300194), characterized by Alport syndrome (ATS) and mental retardation (MR), indicated Xq22.3 as a region containing one mental retardation gene. Comparing the extent of deletion between individuals with ATS MR and individuals with ATS alone allowed us to define a critical region for mental retardation of approximately 380 kb, containing four genes. Here we report the identification of two point mutations, one missense and one splice-site change, in the gene FACL4 in two families with nonspecific mental retardation. Analysis of enzymatic activity in lymphoblastoid cell lines from affected individuals of both families revealed low levels compared with normal cells, indicating that both mutations are null mutations. All carrier females with either point mutations or genomic deletions in FACL4 showed a completely skewed X inactivation, suggesting that the gene influences survival advantage. FACL4 is the first gene shown to be involved in nonspecific mental retardation and fatty acid metabolism. PMID- 11889466 TI - A new type of mutation causes a splicing defect in ATM. AB - Disease-causing splicing mutations described in the literature primarily produce changes in splice sites and, to a lesser extent, variations in exon-regulatory sequences such as the enhancer elements. The gene ATM is mutated in individuals with ataxia-telangiectasia; we have identified the aberrant inclusion of a cryptic exon of 65 bp in one affected individual with a deletion of four nucleotides (GTAA) in intron 20. The deletion is located 12 bp downstream and 53 bp upstream from the 5' and 3' ends of the cryptic exon, respectively. Through analysis of the splicing defect using a hybrid minigene system, we identified a new intron-splicing processing element (ISPE) complementary to U1 snRNA, the RNA component of the U1 small nuclear ribonucleoprotein (snRNP). This element mediates accurate intron processing and interacts specifically with U1 snRNP particles. The 4-nt deletion completely abolished this interaction, causing activation of the cryptic exon. On the basis of this analysis, we describe a new type of U1 snRNP binding site in an intron that is essential for accurate intron removal. Deletion of this sequence is directly involved in the splicing processing defect. PMID- 11889467 TI - Mutations in the human ortholog of Aristaless cause X-linked mental retardation and epilepsy. AB - Mental retardation and epilepsy often occur together. They are both heterogeneous conditions with acquired and genetic causes. Where causes are primarily genetic, major advances have been made in unraveling their molecular basis. The human X chromosome alone is estimated to harbor more than 100 genes that, when mutated, cause mental retardation. At least eight autosomal genes involved in idiopathic epilepsy have been identified, and many more have been implicated in conditions where epilepsy is a feature. We have identified mutations in an X chromosome linked, Aristaless-related, homeobox gene (ARX), in nine families with mental retardation (syndromic and nonspecific), various forms of epilepsy, including infantile spasms and myoclonic seizures, and dystonia. Two recurrent mutations, present in seven families, result in expansion of polyalanine tracts of the ARX protein. These probably cause protein aggregation, similar to other polyalanine and polyglutamine disorders. In addition, we have identified a missense mutation within the ARX homeodomain and a truncation mutation. Thus, it would seem that mutation of ARX is a major contributor to X-linked mental retardation and epilepsy. PMID- 11889468 TI - CREB required for the stability of new and reactivated fear memories. AB - The cAMP-responsive element binding protein (CREB) family of transcription factors is thought to be critical in memory formation. To define the role of CREB in distinct memory processes, we derived transgenic mice with an inducible and reversible CREB repressor by fusing CREBS133A to a tamoxifen (TAM)-dependent mutant of an estrogen receptor ligand-binding domain (LBD). We found that CREB is crucial for the consolidation of long-term conditioned fear memories, but not for encoding, storage or retrieval of these memories. Our studies also showed that CREB is required for the stability of reactivated or retrieved conditioned fear memories. Although the transcriptional processes necessary for the stability of initial and reactivated memories differ, CREB is required for both. The findings presented here delineate the memory processes that require CREB and demonstrate the power of LBD-inducible transgenic systems in the study of complex cognitive processes. PMID- 11889469 TI - Mutant SOD1 causes motor neuron disease independent of copper chaperone-mediated copper loading. AB - Copper-mediated oxidative damage is proposed to play a critical role in the pathogenesis of Cu/Zn superoxide dismutase (SOD1)-linked familial amyotrophic lateral sclerosis (FALS). We tested this hypothesis by ablating the gene encoding the copper chaperone for SOD1 (CCS) in a series of FALS-linked SOD1 mutant mice. Metabolic 64Cu labeling in SOD1-mutant mice lacking the CCS showed that the incorporation of copper into mutant SOD1 was significantly diminished in the absence of CCS. Motor neurons in CCS-/- mice showed increased rate of death after facial nerve axotomy, a response documented for SOD1-/- mice. Thus, CCS is necessary for the efficient incorporation of copper into SOD1 in motor neurons. Although the absence of CCS led to a significant reduction in the amount of copper-loaded mutant SOD1, however, it did not modify the onset and progression of motor neuron disease in SOD1-mutant mice. Hence, CCS-dependent copper loading of mutant SOD1 plays no role in the pathogenesis of motor neuron disease in these mouse models. PMID- 11889471 TI - Neurological intensive care in India--disease spectrum and outcome. AB - In recent years neurological and neurosurgical intensive care (NNICU) has evolved into a well-recognized subspeciality world over. However it is still a novel concept in the developing world. The admission criteria are variable and flexible. The pattern of disease in the NNICU varies according to the admission policy. In the west, cerebrovascular diseases account for a significant proportion of admissions. In a few studies from the developing countries infections of central nervous system are additional causes requiring intensive care. At our center the disease admission pattern was similar to the pattern reported from the developed countries. Predictors of outcome of neurointensive care have not been systematically evaluated. Limited available data concerns patients of neurotrauma. In addition to the disease severity indices, pre existing chronic illness, adverse events during ICU stay, and the 24-hour presence of a physician also influence patient outcome in the NNICU. PMID- 11889470 TI - Leptin inhibits rat hippocampal neurons via activation of large conductance calcium-activated K+ channels. PMID- 11889472 TI - Hemodynamic monitoring in the neurological intensive care unit. AB - The neurointensivist needs to have a thorough understanding of hemodynamic issues and the interaction of the brain and the cardiovascular system. Before one decides to intervene and try to correct an apparent "abnormal hemodynamic parameter" one needs to think whether such an intervention is indeed warranted and what effect the intervention would have on the cerebral circulation. The neurointensivist thus needs to approach these issues differently from the approach an internist or general intensivist would take. PMID- 11889473 TI - Sodium disturbances frequently encountered in a neurologic intensive care unit. AB - Most sodium disturbances in patients with CNS lesions result from disturbed water regulation. Possible systemic and iatrogenic causes must be evaluated prior to treatment. Insufficient secretion of ADH leads to hypernatremia if fluid intake is inadequate and can be treated with either fluid or hormone replacement. Care must be exercised in patients with acute diabetes insipidus because of the potentially variable and transient nature of the disturbance. Hyponatremia usually results from inappropriate secretion of ADH and should be managed aggressively in symptomatic patients with loop diuretics and hypertonic saline. However, very rapid correction or overcorrection should be avoided. Patients with SAH and hyponatremia should not be fluid restricted because of the risk of exacerbating vasospasm but treated with large volumes of isotonic or mildly hypertonic saline. PMID- 11889474 TI - Neurological emergencies--diabetes management. AB - Diabetes can affect the nervous system in several ways. Of all the neurological complications of diabetes, peripheral neuropathy is by far the commonest and has been extensively studied. The involvement of central nervous system can be in several forms. The underlying damage may be due to involvement of the large and small cerebral blood vessels as also due to metabolic derangement caused by prolonged hypoglycemia, anoxia or ketoacidosis. The neurological emergencies that occur in diabetes can be: 1) atherothrombotic and lacunar strokes; 2) convulsive disorder in the setting of both hypo and hyperglycemia; 3) coma; 4) cranial neuropathies; and 5) acute proximal muscle weakness. In patients with diabetes, atherothrombotic stroke is associated with poor outcome. Hyperglycemia at the time of stroke is an important risk factor for an adverse outcome than chronic stable diabetic state. Proper management of diabetes in these acute situations is crucial for a better outcome of the underlying disease process. PMID- 11889475 TI - Diagnosis and management of increased intracranial pressure. AB - Increased intracranial pressure (ICP) is a pathological state common to a variety of neurological diseases, all of which are characterized by the addition of volume to the skull contents. Elevated ICP may lead to brain damage or death by two principle mechanisms: 1) global hypoxic-ischemic injury, as a consequence of reduced cerebral perfusion pressure (CPP) and cerebral blood flow; and 2) mechanical distortion and compression of brain tissue as a result of intracranial mass effect and ICP compartmentalization. All ICP therapies have as a goal, reduction of intracranial volume. In unmonitored patients with acute neurological deterioration, head elevation, hyperventilation, and mannitol (1g/kg) can rapidly lower ICP. Fluid-coupled ventricular catheters and fiberoptic transducers are the most accurate and reliable instruments for measuring ICP. In monitored patients, the treatment of critically raised ICP should proceed in an orderly step-wise fashion: 1) consideration of neuroimaging to exclude a new surgically operable lesion; 2) intravenous sedation to attain a quiet motionless state; 3) manipulation of blood pressure to keep CPP >70 and <120; 4) mannitol infusion; 5) moderate hyperventilation (P(CO2) 26 to 30 mmHg); and 6) high-dose pentobarbital therapy. Application of moderate hypothermia (32 to 33 degrees C) shows promise as a newer method for treating refractory ICP. Placement of an ICP monitor is the critical first step in management of ICP. Treatment is best done using a stepwise protocol, with careful attention to sedation and CPP control prior to using mannitol and hyperventilation. PMID- 11889477 TI - Airway management in critically ill neurological patients. AB - Respiratory complications play an important part in the morbidity and mortality of critically ill neurological patients. Assurance of airway patency is of primary concern in such patients. A plethora of airway maintenance techniques and devices have been recommended for securing and maintaining the airway. But, translaryngeal intubation through the oral route is the safest and most preferred technique. Proper assessment and adequate preparation of the patient before intubation helps to avert crises. In difficult intubation one may secure and maintain the airway by placing a laryngeal mask airway (LMA). The role of early tracheotomy in patients who require prolonged ventilatory support can not be overemphasized. However, the development of inert and softer endotracheal tubes with low pressure cuff has reduced the complications associated with endotracheal intubation. Finally and most importantly the best place to acquire competence in securing the airway is the operation theater not the intensive care unit. PMID- 11889476 TI - Infections in neurology and neurosurgery intensive care units. AB - Nosocomial infections are common among hospitalized patients, more so in intensive care units (ICU). They contribute significantly to morbidity, mortality and cost of care. Few studies address the issue of nosocomial infections in Neurology and neurosurgery ICUs, (NNICU) and data from other ICUs probably cannot be extrapolated to acutely ill neurologic patients. While the incidence of urinary tract infections and catheter related infections may be similar to those in other ICUs, comatose patients may be at a greater risk of nosocomial pneumonia. Certain nosocomial infections are peculiar to NNICU and appear to be associated with higher mortality and morbidity. A systematic approach to evaluation of new episodes of fever, informed use of empirical antibiotics in the context of prevailing drug sensitivities and developing a hospital infection control program are methods crucial to controlling and preventing nosocomial infections. Infections in the intensive care unit (ICU) have been under intense study over the last two decades. Nosocomial infections are common and to a large extent, preventable. However, an established infection by multidrug resistant bacteria is difficult to treat and results in a high mortality, morbidity and cost of care. This article addresses nosocomial infections in the context of the Neurology and Neurosurgery ICU (NNICU). PMID- 11889478 TI - Nutrition in neurologic and neurosurgical critical care. AB - Till recently little attention was paid to the nutritional and metabolic management of the neurologic patient. The effect of brain insult on metabolic and nutritional demands has only recently been studied extensively. Patients with head injury suffer a systemic metabolic response to stress consisting of hypermetabolism/hypercatabolism, hyperglycemia, acute phase response, and immune system alterations. The cause of these abnormalities is probably multifactorial. Resting energy expenditure (REE) increases following closed head injury. Until recently nutritional management was not considered in the medical treatment of these patients. Several studies have proved that adequate nutritional support decreases mortality and morbidity. Enteral nutrition is proved to accelerate normalization of nutritional status and to improve substance tolerance. PMID- 11889480 TI - Severity-of-illness scoring systems and models: neurological and neurosurgical intensive care units. AB - Predicting the outcome of critically ill patients admitted to intensive care units (ICU) has undergone considerable evolution over the last two decades. Various general purpose severity-of-illness scoring systems, Acute Physiology and Chronic Health Evaluation score (APACHE II, APACHE III), Mortality Predicting Model (MPM II), and Simplified Acute Physiology Score (SAPS II), have been extensively validated for large groups of critical care patients with mixed diagnoses and found to correlate well with observed outcome in general. The general hypothesis underlying the use of severity-of-illness scoring systems is that clinical variables that can be assessed on ICU admission and subsequent days of stay in the ICU predict survival and other outcomes of critically ill patients. Variables included in severity-of-illness scoring systems measure specific clinical and physiological functions that are major determinants of mortality. These scoring systems may be used alone within a single disease category or any other independently defined patient group to perform relative risk stratification. Variation in death rates by disease reflects the nature of the underlying process. There is a need for using these scoring systems in specialized ICUs like neurological and neurosurgical ICU as they allow, in addition to predicting outcome, evaluation of new therapies, monitoring of resource utilization and quality assessment of intensive care units. PMID- 11889479 TI - Transcranial Doppler ultrasonography in the neurologic intensive care unit. AB - Transcranial doppler ultrasonography (TCD) is a noninvasive monitoring tool which allows imaging of blood flow velocities in intracranial blood vessels. It is safe, portable, easy to perform and provides accurate information regarding underlying physiology which may help to guide therapy in critically ill neurologic patients. It has significantly contributed to the management of vasospasm related to subarachnoid hemorrhage in the neurologic intensive care unit. TCD is also helpful in the early diagnosis of a variety of complications that can occur in patients with head injury such as vasospasm, elevated intracranial pressure and disordered cerebral autoregulation. Careful performance of the test and experienced interpretation can identify TCD waveforms indicative of cerebral circulatory arrest, an ancillary finding used for the diagnosis of brain death. TCD is likely to play a larger role in evaluation of the patient in the future because of its safety, portability and ability to define moment-to moment changes in cerebral blood flow velocities and cerebral blood flow. PMID- 11889481 TI - The ribulose-1,5-bisphosphate carboxylase/oxygenase gene cluster of Methylococcus capsulatus (Bath). AB - The genes encoding the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Methylococcus capsulatus (Bath) were localised to an 8.3-kb EcoRI fragment of the genome. Genes encoding the large subunit ( cbbL), small subunit ( cbbS) and putative regulatory gene ( cbbQ) were shown to be located on one cluster. Surprisingly, cbbO, a second putative regulatory gene, was not located in the remaining 1.2-kb downstream (3') of cbbQ. However, probing of the M. capsulatus (Bath) genome with cbbO from Nitrosomonas europaea demonstrated that a cbbO homologue was contained within a separate 3.0-kb EcoRI fragment. Instead of a cbbR ORF being located upstream (5') of cbbL, there was a moxR-like ORF that was transcribed in the opposite direction to cbbL. There were three additional ORFs within the large 8.3-kb EcoRI fragment: a pyrE-like ORF, an rnr-like ORF and an incomplete ORF with no sequence similarity to any known protein. Phylogenetic analysis of cbbL from M. capsulatus (Bath) placed it within clade A of the green type Form 1 Rubisco. cbbL was expressed in M. capsulatus (Bath) when grown with methane as a sole carbon and energy source under both copper-replete and copper limited conditions. M. capsulatus (Bath) was capable of autotrophic growth on solid medium but not in liquid medium. Preliminarily investigations suggested that other methanotrophs may also be capable of autotrophic growth. Rubisco genes were also identified, by PCR, in Methylococcus-like strains and Methylocaldum species; however, no Rubisco genes were found in Methylomicrobium album BG8, Methylomonas methanica S1, Methylomonas rubra, Methylosinus trichosporium OB3b or Methylocystis parvus OBBP. PMID- 11889482 TI - Characterization of the urease gene cluster from Rhizobium leguminosarum bv. viciae. AB - Moderate levels of urease activity (ca. 300 mU mg(-1)) were detected in Rhizobium leguminosarum bv. viciae UPM791 vegetative cells. This activity did not require urea for induction and was partially repressed by the addition of ammonium into the medium. Lower levels of urease activity (ca. 100 mU mg(-1)) were detected also in pea bacteroids. A DNA region of ca. 9 kb containing the urease structural genes ( ureA, ureB and ureC), accessory genes ( ureD, ureE, ureF, and ureG), and five additional ORFs ( orf83, orf135, orf207, orf223, and orf287) encoding proteins of unknown function was sequenced. Three of these ORFs ( orf83, orf135 and orf207) have a homologous counterpart in a gene cluster from Sinorhizobium meliloti, reported to be involved in urease and hydrogenase activities. R. leguminosarum mutant strains carrying Tn 5 insertions within this region exhibited a urease-negative phenotype, but induced wild-type levels of hydrogenase and nitrogenase activities in bacteroids. orf287 encodes a potential transmembrane protein with a C-terminal GGDEF domain. A mutant affected in orf287 exhibited normal levels of urease activity in culture cells. Experiments aimed at cross-complementing Ni-binding proteins required for urease and hydrogenase synthesis (UreE and HypB, respectively) indicated that these two proteins are not functionally interchangeable in R. leguminosarum. PMID- 11889483 TI - Purification and characterization of the methylene tetrahydromethanopterin dehydrogenase MtdB and the methylene tetrahydrofolate dehydrogenase FolD from Hyphomicrobium zavarzinii ZV580. AB - Recently, it has been shown that heterotrophic methylotrophic Proteobacteria contain tetrahydrofolate (H(4)F)- and tetrahydromethanopterin (H(4)MPT)-dependent enzymes. Here we report on the purification of two methylene tetrahydropterin dehydrogenases from the methylotroph Hyphomicrobium zavarzinii ZV580. Both dehydrogenases are composed of one type of subunit of 31 kDa. One of the dehydrogenases is NAD(P)-dependent and specific for methylene H(4)MPT (specific activity: 680 U/mg). Its N-terminal amino acid sequence showed sequence identity to NAD(P)-dependent methylene H(4)MPT dehydrogenase MtdB from Methylobacterium extorquens AM1. The second dehydrogenase is specific for NADP and methylene H(4)F (specific activity: 180 U/mg) and also exhibits methenyl H(4)F cyclohydrolase activity. Via N-terminal amino acid sequencing this dehydrogenase was identified as belonging to the classical bifunctional methylene H(4)F dehydrogenases/cyclohydrolases (FolD) found in many bacteria and eukarya. Apparently, the occurrence of methylene tetrahydrofolate and methylene tetrahydromethanopterin dehydrogenases is not uniform among different methylotrophic alpha-Proteobacteria. For example, FolD was not found in M. extorquens AM1, and the NADP-dependent methylene H(4)MPT dehydrogenase MtdA was present in the bacterium that also shows H(4)F activity. PMID- 11889484 TI - Correlation between acetaldehyde and ethanol resistance and expression of HSP genes in yeast strains isolated during the biological aging of sherry wines. AB - In the production of sherry wines, the process of biological aging is essential for the development of their organoleptic properties. This process involves velum formation by "flor" yeasts. Several of these yeast strains have been isolated and characterized with regard to their genetic, physiological and metabolic properties. In this work, we studied their resistance to cold-, osmotic-, oxidative-, ethanol- and acetaldehyde-stress, and found, in most cases, a correlation between resistance to acetaldehyde stress and ethanol stress and isolation from "soleras." Moreover, gene expression analysis revealed induction of the heat shock protein (HSP) genes HSP12, HSP82, and especially HSP26 and HSP104, under acetaldehyde stress in most of the strains. In strain C, there was a clear correlation between resistance to ethanol and acetaldehyde, the high induction of HSP genes by these compounds and its presence as the predominant strain in most levels of several soleras. PMID- 11889485 TI - The sensor kinase CitA (DpiB) of Escherichia coli functions as a high-affinity citrate receptor. AB - For the CitA-CitB (DpiB-DpiA) two-component signal transduction system from Escherichia coli, three diverse functions have been reported: induction of the citrate fermentation genes citCDEFXGT, repression of the regulator gene appY, and destabilization of the inheritance of iteron-containing plasmids such as pSC101. This poses the question of the principal biological role of this system. Here it is shown that the periplasmic domain of the E. coli sensor kinase CitA functions as a high-affinity citrate receptor. Two CitA derivatives were purified by affinity chromatography and subjected to binding studies using isothermal titration calorimetry (ITC). One of them, termed CitA215MBP, comprised the N terminal part of CitA (amino acid residues 1-215), including the two transmembrane helices, and was fused to the amino terminus of the E. coli maltose binding protein lacking its signal peptide. The second CitA derivative, designated CitAP(Ec), encompassed only the periplasmic domain (amino acid residues 38-177). CitA215MBP bound citrate at 25 degrees C with a K(d) of 0.3 microM and a binding stoichiometry of up to 0.9 in 50 mM sodium phosphate buffer, pH 7. Binding was driven by the enthalpy change (Delta H of -95.7 kJ mol(-1)), whereas the entropy change was not favorable for binding ( T Delta S of -58.6 kJ mol(-1)). ITC experiments with CitAP(Ec) yielded similar K(d) values for citrate (0.15-1.0 microM). Besides citrate, also isocitrate ( K(d) approximately tricarballylate ( K(d) approximately t not malate were bound by CitAP(Ec). The results favor the assumption that the primary biological function of the CitA CitB system is the regulation of the citrate fermentation genes. PMID- 11889486 TI - Effect of environmental factors on the synthesis of scytonemin, a UV-screening pigment, in a cyanobacterium (Chroococcidiopsis sp.). AB - Abstract. The UV-screening pigment scytonemin is found in many species of ensheathed cyanobacteria. Past work has shown that the pigment is synthesized in response to exposure to UV-A irradiance. This study investigated the effect of other correlated stress factors including heat, osmotic and oxidative stress on the synthesis of scytonemin in a clonal cyanobacterial isolate ( Chroococcidiopsis sp.) from an epilithic desert crust. Stress experiments were carried out both in conjunction with UV-A irradiance and in isolation. Increases in both temperature and photooxidative conditions in conjunction with UV-A caused a synergistic increase in the rate of scytonemin production. In contrast, increased salt concentration under UV-A irradiance inhibited scytonemin synthesis. However, unlike the responses to temperature and oxidative stress, cells synthesized low levels of scytonemin under osmotic stress in the absence of scytonemin-inducing irradiance. These results suggest that scytonemin induction may be regulated as a part of a complex stress response pathway in which multiple environmental signals affect its synthesis. PMID- 11889487 TI - Characterization of pMa025, a plasmid from the cyanobacterium Microcystis aeruginosa UV025. AB - The characterization of pMa025, a plasmid isolated from the unicellular, toxin producing cyanobacterium Microcystis aeruginosa UV025, is described. A recombinant plasmid, pMaL [pMa025-pBluescript II SK(-)] was constructed for mapping, sequencing, and development of shuttle vectors capable of transforming both Escherichia coli and M. aeruginosa. pMa025 is 8,018 bp in length and has a G+C content of 62.3 mol%. Nineteen presumptive ORFs, ORF A - ORF S were identified using ATG or GTG as initiation codons. Fifteen different ORFs, ORF a - ORF o were identified using TGA as a degenerate codon for tryptophan. GTG was the start codon in two-thirds of the putative ORFs when TGA was the termination codon. GTG was the start codon in one-third of the putative ORFs when TGA was used as a codon for tryptophan. The deduced amino acid sequence from ORF j (3,114 bp) was significantly similar to that of a putative plasmid replication protein, RepA, from plasmid pUH24 of Synecho coccus sp. strain PCC7942. M. aeruginosa UV027 and E. coli were transformed to carbenicillin resistance with pMaL-D7, a 6.4-kb hybrid plasmid (3.46 kb pMa025, 2.95 kb pBluescript II) generated from the nested deletion strategy. pMaL-D7 will be used as a shuttle vector. PMID- 11889488 TI - Thermaerobacter nagasakiensis sp. nov., a novel aerobic and extremely thermophilic marine bacterium. AB - A novel, extremely thermophilic bacterium was isolated from a shallow marine hydrothermal vent at depth of 22 m in Tachibana Bay, Nagasaki Prefecture, Japan. Cells were gram-negative, non-spore-forming, motile rods. Growth was observed between 52 and 78 degrees C (optimum 70 degrees C), pH 5 and 8 (optimum pH 7) and 0-4.5% NaCl (optimum 1.0%). The isolate was a strictly aerobic heterotroph utilizing yeast extract and trypticase peptone. The G+C content of the genomic DNA is 69 mol%. Analysis of 16S rDNA sequences indicated that strain Ts1a is closely related to Thermaerobacter marianensis. The differences in physiology and DNA-DNA similarity between strain Ts1a and T. marianensis showed that strain Ts1a represents a new species of Thermaerobacter. The type strain of T. nagasakiensis is strain Ts1a (=JCM11223, DSM 14512). PMID- 11889490 TI - The SRD2 gene is involved in Saccharomyces cerevisiae morphogenesis. AB - Saccharomyces cerevisiaepresents two alternative vegetative forms of growth, switching between yeast forms to pseudohyphal forms depending on the specific environmental conditions. To identify genes involved in cell wall morphogenesis, a haploid S. cerevisiae monomorphic mutant, W27, which exhibits pseudohyphal growth in the absence of the normal external signals that induce the formation of filamentous forms, was characterized. S. cerevisiaeW27 did not demonstrate agar invasive growth, a characteristic of most filamentous strains. The mutant wall had no obvious alterations with respect to mannan and glucan content, but had three times more chitin than the parental strain. This produced an increase in the amount of proteins linked covalently to chitin. The same protein species, however, were released from the cell walls of the mutant and the parental strain. The W27 mutation was complemented with a genomic library and the SRD2/ECM23 gene was identified as the complementing ORF. Transformation of S. cerevisiaeW27 with the Ycplac33 vector carrying the SRD2 gene produced the original phenotype. These results suggest that the SRD2gene acts as a negative regulator of pseudohyphal growth. PMID- 11889489 TI - Ferredoxin-mediated reactivation of the chlorocatechol 2,3-dioxygenase from Pseudomonas putida GJ31. AB - In the chlorobenzene degrader Pseudomonas putida GJ31, chlorocatechol is formed as an intermediate and cleaved by a meta-cleavage extradiol chlorocatechol dioxygenase, which has previously been shown to be exceptionally resistant to inactivation by substituted catechols. The gene encoding this dioxygenase ( cbzE) is preceded by a gene ( cbzT) potentially encoding a ferredoxin, the function of which was studied. The cbzT gene product was overproduced in Escherichia coli and purified in recombinant form. Two homologous proteins, CdoT and AtdS, encoded by genes identified in strains degrading nitrobenzene and aniline, respectively, were also purified and characterized. All three proteins showed spectroscopic properties typical for [2Fe-2S] ferredoxins. The chlorocatechol dioxygenase from strain GJ31 (CbzE) was fully inactivated when 4-methylcatechol was used as substrate. Inactivated CbzE could be rapidly reactivated in vitro in the presence of purified CbzT and a source of reductant. It is inferred that the ability of strain GJ31 to metabolize both chlorobenzene and toluene might depend on the regeneration of the chlorocatechol dioxygenase activity mediated by CbzT. Three CbzT-like ferredoxins, including AtdS, were found to be competent in the reactivation of CbzE, whereas XylT, a protein known to mediate reactivation of the catechol dioxygenase from P. putida mt2 (XylE), was ineffective. Accordingly, CbzT formed a covalent complex with CbzE when cross-linked with a carbodiimide, whereas XylT did not. In the reverse situation, CbzT was found to reactivate XylE as efficiently as XylT and formed an heterologous covalent complex with this enzyme upon cross-linking. We conclude that CbzT, CdoT and AtdS are isofunctional ferredoxins that appear to be involved in the reactivation of their cognate catechol dioxygenases. Based on primary structure comparisons, residues of the ferredoxins possibly involved in the molecular interaction with catechol dioxygenases were identified and their significance is discussed. PMID- 11889492 TI - Infralimbic kappa opioid and muscarinic M1 receptor interactions in the concurrent modulation of anxiety and memory. AB - RATIONALE: Spontaneous working memory and anxiety-like behaviour can be concurrently influenced following kappa opioid or muscarinic M1 antagonist infusions in the infralimbic (IL) area of the ventromedial prefrontal cortex (vmPFC) in CD-1 mice. Further dose-response analyses of our previous norBNI and pirenzepine data revealed significant dose x drug interactions on trial-1 and -2 anxiety-related elevated plus-maze indices. These data prompted us to evaluate the effects of simultaneous IL norBNI/pirenzepine infusions on anxiety and spontaneous working memory. OBJECTIVE: The present study sought to evaluate whether (a) our previously reported anxiogenic and working memory disruptive effects of norBNI, and anxiolytic and working memory disruptive effects of pirenzepine data could be replicated using the most effective dose (10 nmol) of each drug and (b) IL infusions of mixed kappa/M1 receptor inhibitor drugs might interactively influence these cognitive, behavioural processes. METHODS: Anxiety was evaluated in the elevated plus maze, and spontaneous alternation memory was evaluated in the Y-maze following pirenzepine, norBNI or two levels of norBNI/pirenzepine drug mix infusions in the IL vmPFC. RESULTS: Pretreatment with the M1 antagonist pirenzepine was anxiolytic in trial 1 (10 nmol) and trial 2 (no injection) in the elevated plus maze 24 h later, and disrupted alternation performance and some aspects of attention in the Y-maze. Pretreatment with the kappa antagonist norBNI was anxiogenic in trial 1 (10 nmol) and trial 2 (no injection) in the elevated plus maze 24 h later, and also disrupted alternation performance and some aspects of attention in the Y-maze. The norBNI-10 nmol/pirenzepine-10 nmol mixed drug infusion was somewhat anxiogenic in trial 1, exerted no carry-over effects in trial 2 in the elevated plus maze, and disrupted alternation memory and some aspects of attention in the Y-maze. The norBNI-5 nmol/pirenzepine-10 nmol drug mix had no effect on trial-1 or -2 anxiety measures in the elevated plus maze, yet also disrupted Y-maze spontaneous memory performance. CONCLUSIONS: (1) The effects of IL infusions of norBNI or pirenzepine (10 nmol/0.5 microl) alone on anxiety-like behaviour and aversive learning and memory in the elevated plus-maze replicated previously reported data. (2) Mixed M1/kappa receptor inhibition in the IL cortex exerted counteractive effects on anxiety-like behaviour and aversive learning in the elevated plus maze. (3) Mixed M1/kappa receptor inhibition appeared to exert additive disruptive effects on alternation performance and aspects of attention related to active working memory in the Y-maze. PMID- 11889491 TI - Conditioning tastant and the acquisition of conditioned taste avoidance to drugs of abuse in DBA/2J mice. AB - RATIONALE: Conditioned taste aversion (CTA) produced by drugs of abuse such as morphine and cocaine has been interpreted as representing the rewarding actions of these drugs. Evidence for this interpretation is based, in part, on findings in rats indicating saccharin is a more effective conditioning flavor compared to salt (NaCl). However, our studies with ethanol have found salt to be a highly effective conditioning flavor in mice. OBJECTIVES: The present series of studies examined the acquisition of CTA to morphine, ethanol, lithium chloride, and cocaine. Further, saccharin and salt were utilized in each experiment in order to determine effectiveness of each flavor to serve as a conditioning stimulus. METHODS: In four separate experiments, adult male DBA/2J mice were acclimated to a 2 h/day water restriction regimen. Subsequently they received four conditioning trials consisting of 1 h access to either 0.15% w/v saccharin or 0.1 M salt followed by 0, 10 or 20 mg/kg morphine (experiment 1), 0, 2, or 4 g/kg ethanol (experiment 2), 0, 1.5 or 3.0 milliequivalents/kg lithium chloride (experiment 3) or 0, 10 or 20 mg/kg cocaine (experiment 4). A fifth flavor access period (trial 5) was not followed by drug exposure. Following trial 5, each subject received 24 h access to the conditioning flavor and water (two-bottle test 1). Control subjects (0 dose groups from each experiment) received a second two-bottle test with 24-h access to both saccharin and salt flavors. RESULTS: Reduced flavor intake and reduced flavor preference was noted in all drug-paired groups in each experiment. However, more rapid development of CTA was seen with the saccharin flavor in morphine- or cocaine-paired groups. In contrast, ethanol-induced CTA appeared more rapidly with the salt flavor. Lithium-induced CTA was modest, and emerged equally with either flavor. CONCLUSIONS: CTA induced by morphine or cocaine in mice occurs in a similar pattern to that seen in rats, and these findings agree with an interpretation based on drug reward. In contrast, ethanol induced CTA is more likely attributable to aversive effects. PMID- 11889493 TI - Provision of cues to signal a withdrawal US prevents the US pre-exposure effect in a diazepam-withdrawal conditioned taste aversion. AB - RATIONALE: Prior experience of withdrawal from chronic diazepam treatment reduces the aversiveness of withdrawal when precipitated withdrawal is made the US in a conditioned taste aversion (CTA) paradigm. Some accounts of US pre-exposure reducing its effectiveness in CTA postulate that US pre-exposure leads to the formation of associations with the environment, resulting in blocking of taste conditioning, but a test of a blocking explanation failed to provide support for such an account. OBJECTIVE: The present experiments tested alternative explanations. METHODS: Male mice (C57BLx129sv derived) made dependent upon diazepam (15 mg/kg per day, SC) were subjected to precipitated withdrawal with IP flumazenil (20 mg/kg) as the unconditioned stimulus (US) in a conditioned taste aversion (CTA) paradigm, in which sucrose was the conditioned stimulus (CS). The conditioning trial took place in either the same or different environment from that in which the mice had received pre-exposure to the withdrawal US. RESULTS: No evidence was found that a place conditioned to withdrawal was capable of supporting a second-order CTA. A second experiment showed that whether withdrawal supported a CTA depended upon whether the previous experience of withdrawal had been predicted by an environmental stimulus. CONCLUSIONS: Prior experience of an unpredictable aversive US disrupts the subsequent formation of a CTA to the same US. Since prior experience of withdrawal, if it was predictable by an environmental event, did not prevent withdrawal from being a US in a subsequent CTA experiment, withdrawal retains its aversive nature even following prior experience. An explanation in terms of the nature of US predictability following repeated withdrawal from diazepam is consistent both with the current data and our previous findings. PMID- 11889494 TI - Differential muscarinic and NMDA contributions to visuo-spatial paired-associate learning in rhesus monkeys. AB - RATIONALE: Early, accurate detection of degenerative neurological disorders such as Alzheimer's disease (AD) is essential for therapies designed to slow disease progression. Performance of a touch-screen mediated visuo-spatial paired associates learning (vsPAL) task predicts neurocognitive decline in elderly populations presenting with mild cognitive impairment and distinguishes AD patients from elderly depressed individuals. Translation of this cognitive task to a non-human model may therefore provide an improved tool for study of the etiology and treatment of dementia. OBJECTIVE: The goal of the current study was to contrast cholinergic and glutamatergic contributions to performance of this AD sensitive task by challenging rhesus monkeys performing vsPAL with muscarinic antagonist and non-competitive NMDA antagonist drugs. METHODS: Seven monkeys were trained to perform vsPAL and then serially challenged with acute doses of scopolamine (3, 10, 17 microg/kg, IM) and ketamine (0.3, 1.0, 1.78 mg/kg, IM). RESULTS: Scopolamine produced a dosexdifficulty related impairment of both recognition memory and incremental acquisition aspects of task performance. In contrast, ketamine administration resulted in a dose-dependent impairment of recognition memory but not incremental acquisition. CONCLUSIONS: Monkeys' performance of a task sensitive to AD in humans was impaired by two classic pharmacological models of cognitive impairment, therefore supporting the use of this nonhuman model to explore mechanisms of AD-associated cognitive decline. The differential pattern of impairment observed is consistent with a hypothesis that muscarinic mechanisms are required for linking external events with an existing internal representation, whereas NMDA mechanisms are required for the formation/strengthening of such an internal representation. PMID- 11889495 TI - Effects of GBR 12909, WIN 35,428 and indatraline on cocaine self-administration and cocaine seeking in rats. AB - RATIONALE: It has been proposed that drugs that decrease cocaine self administration or cocaine seeking by laboratory animals might be effective pharmacotherapies in the treatment of cocaine addiction. Previous studies have suggested that the dopamine uptake inhibitor, GBR 12909, might be such a candidate drug because it decreases cocaine self-administration. Other studies have shown that GBR 12909 elicits cocaine seeking, which might limit its utility as an anti-cocaine pharmacotherapy. OBJECTIVES: The present study sought to compare the potency of GBR 12909 in decreasing cocaine self-administration and in eliciting cocaine seeking. These findings were compared to effects produced by the cocaine analog, WIN 35,428 and the non-specific monoamine uptake inhibitor, indatraline. METHODS: A within-session protocol was used to obtain the dose effect relationship for cocaine self-administration in a single 5-6 h daily test session. Rats were pretreated with GBR 12909 (3.0-30.0 mg/kg), WIN 35,428 (0.1 1.0 mg/kg) or indatraline (0.03-1.00 mg/kg) 30 min prior to the test session. RESULTS: GBR 12909 and WIN 35,428 decreased responding maintained by intermediate and high doses of cocaine but indatraline failed to alter the cocaine dose-effect curve. Other groups of rats demonstrated that pretreatment with all three drugs reinstated extinguished cocaine-taking behavior, although indatraline was less efficacious than either GBR 12909 or WIN 35,428. Cocaine-produced drug seeking was enhanced by pretreatment with the highest doses of GBR 12909 and WIN 35,428 but was unaffected by pretreatment with indatraline. A low dose of GBR 12909 that decreased cocaine self-administration failed to produce drug seeking but equal doses of WIN 35,428 were required to decrease cocaine self-administration and to elicit drug seeking. CONCLUSIONS: These data suggest a preferred profile of effects of GBR 12909 as an anti-cocaine pharmacotherapy. It is noted, however, that this drug might be expected to potentiate the ability of cocaine to elicit cocaine seeking following abstinence. PMID- 11889496 TI - Ovarian steroid regulation of monoamine oxidase-A and -B mRNAs in the macaque dorsal raphe and hypothalamic nuclei. AB - RATIONALE: The serotonin neural system plays a pivotal role in mood, affective regulation and integrative cognition, as well as numerous autonomic functions. We have shown that ovarian steroids alter the expression of several genes in the dorsal raphe of macaques, which may increase serotonin synthesis and decrease serotonin autoinhibition. Another control point in aminergic neurotransmission involves degradation by MAO. This enzyme occurs in two isoforms, A and B, which have different substrate preferences. OBJECTIVES: We questioned the effect of ovarian steroid hormones on MAO-A and MAO-B mRNA expression in the dorsal raphe nucleus and hypothalamus using in situ hybridization in non-human primates. METHODS: Rhesus monkeys ( Macaca mulatta; n=5/group) were spayed and either placebo treated (controls), estrogen (E) treated (28 days), progesterone (P) treated (14 days placebo+14 days P), or E+P treated (14 days E+14 days E+P). Perfusion-fixed sections (25 microm) were hybridized with a 233 bp MAO-A, or a 373 bp MAO-B, radiolabeled-antisense monkey specific probes. Autoradiographic films were analyzed by densitometry, which was performed with NIH Image Software. RESULTS: MAO-A and -B mRNAs were detected in the dorsal raphe nucleus (DRN) and in the hypothalamic suprachiasmatic nucleus (SCN), preoptic area (POA), paraventricular nucleus (PVN), supraoptic nucleus (SON), lateral hypothalamus (LH) and ventromedial nucleus (VMN). MAO-A mRNA optical density was significantly decreased by E, P, and E+P in the DRN and in the hypothalamic PVN, LH and VMN. Ovarian hormones had no effect on MAO-B mRNA expression in the DRN. However, there was a significant decrease in MAO-B optical density in the hypothalamic POA, LH and VMN with E, P or E+P treatment. Pixel area generally reflected optical density. CONCLUSIONS: Ovarian steroids decreased MAO-A, but not B, in the raphe nucleus. However, both MAO-A and B were decreased in discrete hypothalamic nuclei by hormone replacement. These data suggest that the transcriptional regulation of MAO by ovarian steroids may play a role in serotonin or catecholamine neurotransmission and hence, mood, affect or cognition in humans. PMID- 11889497 TI - The selective serotonin re-uptake inhibitors fluvoxamine and paroxetine differ in sexual inhibitory effects after chronic treatment. AB - RATIONALE: The selective serotonin re-uptake inhibitors (SSRIs) delay orgasm and ejaculation in men. In men with rapid ejaculation it was shown that, of the SSRIs, paroxetine exerted the strongest delay in ejaculation and fluvoxamine the weakest. OBJECTIVES: In the present study, we compared the acute and chronic effects of fluvoxamine and paroxetine on sexual behavior in the male rat in order to compare their differential inhibitory effects on sexual behavior. METHODS: During a 4-week period, 48 male Wistar rats, selected on the basis of their sexual performance, were repeatedly tested for sexual behavior. All male rats received vehicle (saline, n=12), fluvoxamine (30 mg/kg, n=12), or paroxetine (10 mg/kg, n=12) daily for 2 weeks. Sexual behavioral tests were performed on days 1 (acute), 7, and 14. RESULTS: After acute oral administration, fluvoxamine and paroxetine did not inhibit sexual behavior. After 7 days and 14 days treatment, fluvoxamine mildly inhibited certain parameters of sexual behavior but ejaculation was never delayed. In contrast, paroxetine, after 7 days and particularly after 14 days treatment, strongly inhibited sexual behavior, including ejaculation. CONCLUSIONS: These results strongly concur with clinical data, suggesting that paroxetine, but not fluvoxamine, delays ejaculation. Because fluvoxamine does not delay ejaculation it may serve as an optimal treatment for depressive illness when sexual side effects, such as a delayed ejaculation, are undesired. The mechanisms whereby paroxetine and fluvoxamine, both being selective serotonin uptake inhibitors, differentially inhibit sexual behavior are unclear. PMID- 11889498 TI - Effects of lesions of the orbitofrontal cortex on sensitivity to delayed and probabilistic reinforcement. AB - RATIONALE: Lesions of the orbital prefrontal cortex (OPFC) can cause pathologically impulsive behaviour in humans. Inter-temporal choice behaviour (choice between reinforcers differing in size, delay and/or probability) has been proposed as a model of "impulsive choice" in animals. OBJECTIVE: The effect of lesions of the OPFC on rats' inter-temporal choice behaviour was examined in two experiments: (1) rats chose between a smaller immediate reinforcer and a larger delayed reinforcer; (2) rats chose between a smaller certain reinforcer and a larger probabilistic reinforcer. METHODS: Under halothane anaesthesia, rats received injections of the excitotoxin quinolinate into the OPFC (0.1 M, 0.5 microl, two injections in each hemisphere), or sham lesions (injections of vehicle). They were trained to press two levers (A and B) for food-pellet reinforcers in discrete-trials schedules. In free-choice trials, a press on A resulted in immediate delivery of one food pellet; a press on B resulted in delivery of two pellets, either following a delay ( d) (experiment 1), or with a probability ( p) <1 (experiment 2). The values of d and p were manipulated across phases of the experiments. The locations of the lesions were verified histologically at the end of the experiment. RESULTS: In experiment 1, both groups showed declining choice of lever B as a function of d. The lesioned rats showed significantly shorter indifference delays ( D50: the value of d corresponding to 50% choice of lever B) than the sham-lesioned rats. In experiment 2, both groups showed declining choice of lever B as a function of the odds against delivery of the two-pellet reinforcer, theta ( theta =[1/ p]-1). The lesioned rats showed lower indifference odds ( theta50: the value of theta corresponding to 50% choice of lever B) than the sham-lesioned rats. In both experiments, the lesioned rats showed extensive atrophy of the OPFC, with sparing of the dorsolateral prefrontal cortex. CONCLUSIONS: The results show that lesions of the OPFC can promote preference for the smaller and more immediate, and the smaller and more certain of two reinforcers. The results are consistent with two interpretations: the lesion may have altered (i) the rates of delay and odds discounting, and/or (ii) sensitivity to the ratio of the sizes of the two reinforcers. PMID- 11889499 TI - Implicit and explicit learning in schizophrenics treated with olanzapine and with classic neuroleptics. AB - OBJECTIVE: Novel and classic neuroleptics differ in their effects on limbic striatal/nucleus accumbens (NA) and prefrontal cortex (PFC) dopamine turnover, suggesting differential effects on implicit and explicit learning as well as on anhedonia. The present study investigates whether such differences can be demonstrated in a naturalistic sample of schizophrenic patients. METHODS: Twenty five inpatients diagnosed with DSM-IV schizophrenic psychosis and treated for at least 14 days with the novel neuroleptic olanzapine were compared with 25 schizophrenics taking classic neuroleptics and with 25 healthy controls, matched by age and education level. PFC/NA-dependent implicit learning was assessed by a serial reaction time task (SRTT) and compared with cerebellum-mediated classical eye-blink conditioning and explicit visuospatial memory. Anhedonia was measured with the Snaith-Hamilton-Pleasure Scale (SHAPS). RESULTS: Implicit (SRTT) and psychomotor speed, but not explicit (visuospatial) learning were superior in the olanzapine-treated group as compared to the patients on classic neuroleptics. Compared to healthy controls, olanzapine-treated schizophrenics showed similar implicit learning, but reduced explicit (visuospatial) memory performance. Acquisition of eyeblink conditioning was not different between the three groups. There was no difference with regard to anhedonia and SANS scores between the patients. CONCLUSION: Olanzapine seems to interfere less with unattended learning and motor speed than classical neuroleptics. In daily life, this may translate into better adaptation to a rapidly changing environment. The effects seem specific, as in explicit learning and eyeblink conditioning no difference to classic NL was found. PMID- 11889500 TI - Differential effects of the dopamine D2/D3 receptor antagonist sulpiride on self administration of morphine into the ventral tegmental area or the nucleus accumbens. AB - RATIONALE: The involvement of dopamine neurotransmission in opiate reward remains controversial. OBJECTIVE: To investigate the dopaminergic basis of opiate reward by comparing the effect of systemic injection of the D2/D3 antagonist sulpiride on morphine self-administration (ICSA) into the ventral tegmental area (VTA) or the nucleus accumbens (NAc) METHODS: BALB/c mice were unilaterally implanted with a guide cannula 1.5 mm above either the VTA or the NAc. On experimental days, a stainless-steel injection cannula was inserted via the guide cannula, and mice were trained to discriminate the arm of a Y-maze reinforced by intracranial morphine microinjections (6.5 pmol or 65 pmol/50 nl) from the neutral arm (no injection). Following acquisition of morphine ICSA, the dopamine D2/D3 receptor antagonist sulpiride (50 mg/kg, i.p.) was administered 30 min before testing. RESULTS: Sulpiride produced an extinction of intra-VTA, but not intra-NAC, morphine self-administration. Extinction in VTA subjects was followed by a re appearance of ICSA, although mice continued to receive sulpiride injections. Extinction was re-induced when the dose of sulpiride was raised to 100 mg/kg, whereas no effect of this dose was detected on intra-NAc self-administration. CONCLUSION: Maintenance of intra-VTA, but not intra-NAc, morphine self administration depends acutely on D2/D3 receptors. However, the deleterious effect of sulpiride on intra-VTA morphine self-administration is transient. Reappearance of ICSA under neuroleptic treatment in VTA subjects may be related to the sensitization effect of intra-VTA morphine infusions, combined with an upregulation of D2/D3 receptors and alterations of DA metabolism by repeated sulpiride injections. PMID- 11889501 TI - Effect of TV3326, a novel monoamine-oxidase cholinesterase inhibitor, in rat models of anxiety and depression. AB - RATIONALE: A high incidence of depression is found in subjects with Alzheimer's disease (AD), in whom many antidepressants are contraindicated because they have anticholinergic activity. We have designed a new cholinesterase inhibitor TV3326 [( N-propargyl-(3R) aminoindan-5-yl)-ethyl methyl carbamate] for the treatment of AD, which has neuroprotective activities and also blocks monoamine oxidase (MAO) A and B in the brain but not in the intestine after chronic administration. OBJECTIVES: To examine the antidepressant and anxiolytic potential of TV3326 in rats and compare them with those of its R isomer TV3279, which lacks MAO inhibitory activity, and of amitriptyline and moclobemide. METHODS: Each of the drugs was administered orally, acutely or once daily for 2 weeks, and its effect was evaluated on the behavior of rats in the forced swim test (FST) and plus maze (EPM) test. RESULTS: Immobility in the FST was reduced by 56% after acute and chronic administration of amitriptyline (10 mg/kg) and by 42% after acute administration of moclobemide (20 mg/kg) and by 63% when this drug was given chronically. TV3326 (26 mg/kg) only reduced immobility (by 44%) when given chronically and inhibited brain MAO-A and -B by more than 66%. TV3279 had no significant effect in the FST. All the drugs except TV3326 increased anxiogenic activity in rats in EPM, as indicated by a more than 50% decrease in the time in open arms after chronic administration. CONCLUSIONS: TV3326 has potential antidepressant-like activity when given in a dose regimen that causes significant inhibition of brain MAO-A and -B. Together with its neuroprotective properties, this action could make TV3326 a potentially valuable drug for the treatment of dementia in patients with depression. PMID- 11889502 TI - Pharmacological manipulation of brain galaninergic system and sexual behavior in male mice. AB - RATIONALE: Available data suggest a complex role for the brain galaninergic system in male sexual behavior; however, the results so far obtained in animals with either galanin or galanin antagonists are conflicting. OBJECTIVE: To define the better influence of galanin on male sexual behavior by studying, in mice, (i) the effect of galanin and of the chimeric galanin peptide M40 on the copulatory performance, and (ii) galanin mRNA levels in hypothalamic arcuate and dorso medial nuclei. METHODS: For the behavioral testing, only sexually sluggish male mice were used. Galanin mRNA levels were studied in both sexually potent and impotent mice by means of in situ hybridization. Standard behavioral parameters for sexual behavior were recorded or calculated. Synthetic galanin (0.05, 0.1 or 1 microg/mouse) and M40 (5 or 20 microg/mouse) were intracerebroventricularly (ICV) injected, 15 min before the copulatory test. Galanin mRNA levels were evaluated. RESULTS: In sexually sluggish male mice, both galanin (0.1 and 1 microg/mouse ICV) and M40 (20 microg/mouse ICV), significantly increased intromission frequency and ejaculation latency; M40 also improved copulatory efficacy. On the other hand, in the hypothalamic arcuate and dorso-medial nuclei, the levels of galanin mRNA were not significantly different in sexually potent and impotent male mice. CONCLUSIONS: These results show that in sexually sluggish male mice the ICV injection of either galanin or the chimeric analogue M40 greatly prolongs the duration of the copulation; without a reduction of the sexual drive or of the copulatory performance. On the other hand, the hybridization experiments seem to rule out an important physiological role of the brain galaninergic system in the regulation of male sexual behavior, at least in mice. PMID- 11889503 TI - Enhancement of the discriminative stimulus effects of phencyclidine by the tetracycline antibiotics doxycycline and minocycline in rats. AB - RATIONALE: Tetracycline antibiotics share some neuroprotective and CNS effects with N-methyl- D-aspartate (NMDA) receptor antagonists. OBJECTIVES: The acute effects of two tetracycline antibiotics were compared to those of the prototypic NMDA antagonist phencyclidine (PCP). METHODS: The effects of minocycline (10-56 mg/kg) and doxycycline (10-56 mg/kg) were evaluated in Sprague-Dawley rats trained to discriminate 2.0 mg/kg IP of PCP from saline under a fixed ratio schedule of food presentation. RESULTS: Even though minocycline and doxycycline did not substitute for PCP, pretreatment with 32 mg/kg of either drug produced leftward shifts in the PCP dose-response curve. The 32 mg/kg dose of minocycline also produced a leftward shift in the dose-response curve for dizocilpine (MK 801), another NMDA channel blocker, in the same subjects. CONCLUSIONS: Tetracycline antibiotics may interact either directly or indirectly with NMDA receptors. This suggests that they might be utilized in treatment of brain disorders in which NMDA receptor over-activation has been implicated. PMID- 11889504 TI - Phobic postural vertigo. Body sway during visually induced roll vection. AB - Patients with phobic postural vertigo (PPV) often report a particularly increased unsteadiness when looking at moving visual scenes. Therefore, the differential effects of large-field visual motion stimulation in roll plane on body sway during upright stance were analyzed in 23 patients with PPV, who had been selected for the integrity of their vestibular and balance systems, and in 17 healthy subjects. Visual motion stimulation induced a sensation of apparent body motion (roll vection) in all patients and normal subjects. Normal subjects showed an increased lateral sway path with a lateral shift of the center of pressure (COP) in stimulus direction (mean 1.67 cm, SD 1.63). The patients also exhibited an increase in sway path during visual motion stimulation; however, their body sway differed from that of normals in that there was no lateral displacement of COP (mean 0.19 cm, SD 0.73). The lateral displacement of COP and the increase in RMS of body sway during visual motion stimulation were significantly greater in normals than in the patients ( p<0.05). The patients' increased body sway without COP deviation does not imply an increased risk of falling. Two explanations are conceivable for this increased body sway without body deviation in patients with PPV: (a) the patients rely more on proprioceptive and vestibular rather than on visual cues to regulate upright stance; or (b) they depend on visual, vestibular, and proprioceptive information, but the threshold at which they initiate a compensatory body sway opposite in direction to a perceived body deviation is lower than in normal subjects. The data support the second explanation. PMID- 11889505 TI - Comparison of brain activity during different types of proprioceptive inputs: a positron emission tomography study. AB - It has been shown that the primary and secondary somatosensory cortex, as well as the supplementary motor area (SMA), are involved in central processing of proprioceptive signals during passive and active arm movements. However, it is not clear whether different cortical areas are involved in processing of different proprioceptive inputs (skin, joint, muscle receptors), what their relative contributions might be, where kinesthetic sensations are formed within the CNS, and how they interact when the full peripheral proprioceptive machinery acts. In this study we investigated the representation of the brain structures involved in the perception of passive limb movement and illusory movement generated by muscle tendon vibration. Changes in cortical activity as indicated by changes in regional cerebral blood flow (rCBF) were measured using positron emission tomography (PET). Twelve subjects were studied under four conditions: (1) passive flexion-extension movement (PM) of the left forearm; (2) induced illusions of movements (VI) similar to the real PM, induced by alternating vibration of biceps and triceps tendons (70-80 Hz) at the elbow; (3) alternating vibration of biceps and triceps tendons (with 20-50 Hz) without induced kinesthetic illusions (VN); and (4) rest condition (RE). The results show different patterns of cortex activation. In general, the activation during passive movement was higher in comparison with both kinds of vibration, and activation during vibrations with induced illusions of movement was more prominent than during vibrations without induced illusions. When the PM condition was contrasted with the other conditions we found the following areas of activation -- the primary motor (MI) and somatosensory area (SI), the SMA and the supplementary somatosensory area (SSA). In conditions where passive movements and illusory movements were contrasted with rest, some temporal areas, namely primary and associative auditory cortex, were activated, as well as secondary somatosensory cortex (SII). Our data show that different proprioceptive inputs, which induce sensation of movement, are associated with differently located activation patterns in the SI/MI and SMA areas of the cortex. In general, the comparison of activation intensities under different functional conditions indicates the involvement of SII in stimulus perception generation and of the SI/MI and SMA areas in the processing of proprioceptive input. Activation of the primary and secondary auditory cortex might reflect the interaction between somatosensory and auditory systems in movement sense generation. SSA might also be involved in movement sense generation and/or maintenance. PMID- 11889506 TI - Non-nociceptive upper limb afferents modulate masseter muscle EMG activity in man. AB - Recent electrophysiological data obtained in anaesthetized rats evidenced jaw muscle excitatory responses to the electrical stimulation of type II limb somatosensory afferents. In the present work, we describe an inhibitory reflex evoked in human masseter muscles by stimulation of non-nociceptive fibres travelling in the median and radial nerves (MED and RAD, respectively). Eighteen healthy volunteers participated in the study. Subjects were seated on a comfortable chair, with the complex head-mandible-neck-trunk and the limbs securely fixed to the chair. Attempts were made to minimize possible interferences due to the activation of afferents other than the stimulated ones. The subjects were instructed to contract masseter muscles at a submaximal level and to maintain a stable level of muscle contraction during all trials. EMG voluntary activity was recorded from both masseter muscles by means of coaxial needle electrodes before and after the electrical stimulation of MED and/or RAD at intensities below pain threshold. In all subjects, MED stimulation induced bilaterally a marked depression of masseter EMG activity, which occurred at a latency of 23.6 +/- 2.1 ms and lasted 27.8 +/- 6.6 ms. RAD stimulation also induced a marked reduction in masseter EMG activity, but this effect was clearly observed in 9 out of 18 subjects, and it showed latency (30.2 +/- 7.5 ms) and duration (44.9 +/- 5.4 ms) significantly longer in comparison with the MED induced effect. All subjects exhibited the inhibitory period in masseter EMG following the simultaneous stimulation of both nerves; this one appeared at a latency not significantly different (25.3 +/- 5.9 ms) and lasted much more (37.4 +/ - 8.2 ms) than EMG depression evoked by MED stimulation. The duration of masseter muscle inhibition, induced by MED and/or RAD stimulation, was inversely related to the level of EMG activity, while latency was not related to it. Significant increases in the inhibitory period duration were also observed by increasing stimulus intensity, within a subthreshold range for the activation of nociceptive fibres. In all cases, the inhibitory period was followed by a later excitatory rebound activity, whose latency and duration depended on the duration of the preceding EMG inhibition and on the background level of masseter activation. In conclusion, results evidenced that the activation of arm somatosensory fibres modulates masseter muscle activity in normal man. This might lead to a coordination between limb and masticatory muscle activity, which is required in several complex motor acts. PMID- 11889507 TI - GABA-inactivation attenuates colinear facilitation in cat primary visual cortex. AB - Neurons in primary visual cortex (V1) respond preferentially to stimuli of a particular orientation falling within a circumscribed region of visual space known as their receptive field (RF). However, the response to an optimally oriented stimulus presented within the RF can be enhanced by the simultaneous presentation of co-oriented, co-linearly aligned flank stimuli falling outside the RF which, when presented alone, fail to activate the cell. This type of contextual effect, termed colinear facilitation, presumably forms the physiological substrate for the integration of the line elements of a contour and the perceptual saliency of a contour in a complex environment. Here we show that colinear facilitation in single cells of cat area V1 can be substantially reduced or abolished by focal inactivation of laterally remote cells in the same area which respond strongly to the co-oriented, colinear flank stimulus inducing the facilitatory effect. The results provide evidence that horizontal intrinsic connections between cells with co-oriented and co-linearly aligned RFs make a major contribution to colinear facilitation in V1. They imply that the neuronal circuitry underlying contour integration and saliency is already present at the earliest stage of visual cortical information processing. PMID- 11889508 TI - Non-linear interaction of angular and translational vestibulo-ocular reflex during eccentric rotation in the monkey. AB - Eccentric sinusoidal rotation with the nose facing out or in leads to gain modulation of the vestibulo-ocular reflex (VOR), which is a result of an interaction between angular and translational VOR. There are conflicting reports with regard to the type of interaction. Combined angular and translational VOR during eccentric sinusoidal rotations over a wide range of target distances (12 180 cm), eccentricities (centric, 30 and 50 cm nose-out and nose-in eccentric) and frequencies (0.1-4 Hz) were studied in macaque monkeys trained to fixate earth-stationary light-emitting diode (LED) targets while binocular eye positions were measured using magnetic search coils. The monkeys were also exposed to sudden unpredictable position steps with peak accelerations of 500 degrees/s(2) using similar eccentricities and target distances. VOR gain enhancement during nose-out eccentric sinusoidal rotation was almost compensatory when the target was visible and was independent of stimulus frequency. Mean responses were still close to ideal when the target was extinguished; however, individual data showed increased variability. Sensitivities of the translational portion of the combined VOR were compensatory. These sensitivities were clearly reduced during nose-in eccentric sinusoidal rotation. Thus, especially for close targets at 4 Hz combined VOR was not compensatory, independent of target visibility. VOR elicited by sudden position steps showed a sequential response: (1) purely angular VOR (up to 40-45 ms); (2) additional translational VOR that was not modulated by target distance (45-65 ms); and (3) translational VOR weighted for target location (>65 ms). We conclude that angular and translational VOR have different latencies during transient accelerations and interact differently during agonistic (nose out) and antagonistic stimulation (nose-in). PMID- 11889509 TI - Anticipatory control of center of mass and joint stability during voluntary arm movement from a standing posture: interplay between active and passive control. AB - Anticipatory control of upright posture is the focus of this study that combines experimental and modeling work. Individuals were asked to raise or lower their arms from two initial postures such that the final posture of the arm was at 90 degrees with respect to the body. Holding different weights in the hand varied the magnitude of perturbation to postural stability generated by the arm movement. Whole body kinematics and ground reaction forces were measured. Inverse dynamic analysis was used to determine the internal joint moments at the shoulder, hip, knee and ankle, and reaction forces at the shoulder. Center of mass (COM) of the arm, posture (rest of the body without the arms) and whole body (net COM) were also determined. Changes in joint moment at the hip, knee and ankle revealed a significant effect of the direction of movement. The polarities of the joint moment response were appropriate for joint stabilization. Net COM change showed a systematic effect of the direction of movement even though the arm COM was displaced by the same amount and in the same direction for both arm raising and lowering conditions. In order to determine the effects of the passive forces and moments on the posture COM, the body was modeled as an inverted pendulum. The model was customized for each participant; the relevant model parameters were estimated from data obtained from each trial. The ankle joint stiffness and viscosity were adjusted to ensure postural equilibrium prior to arm movement. Joint reactive forces and moments generated by the arm movements were applied at the shoulder level of this inverted pendulum; these were the only inputs and no active control was included. The posture COM profile from the model simulation was calculated. Results show that simulated posture COM profile and measured posture COM profile are identical for about 200 ms following the onset of arm movement and then they deviate. Therefore, the initial control of COM is passive in nature and the observed joint moment response is for joint stabilization and not for the control of COM. PMID- 11889510 TI - Control of manipulative forces during unimanual and bimanual tasks in patients with Huntington's disease. AB - The aim of the study was to investigate grip-load force regulation in Huntington's disease (HD) patients as compared to control subjects during the performance of a manipulative task that required rhythmical unimanual or bimanual isodirectional/non-isodirectional actions in the sagittal plane. Results showed that the profile of grip-load ratio force was characterized by maxima and minima that were attained at upward and downward hand positions, respectively. Minimum force ratio was higher in patients than in controls, which points to an elevated baseline that may be related to the inherent bradykinesia observed in HD. Maximum force ratio was also increased in patients, but this effect depended on the performance condition, with largest amplifications occurring during non isodirectional movements. The latter rescaling may be associated with the complexity of the coordination mode and its asymmetrical load characteristics. In addition, the temporal delay between the grip and load force peaks was augmented in patients versus controls, indicating a disturbed coupled activation of both forces. Furthermore, the interval was largest during non-isodirectional movements followed by isodirectional and unimanual movements, which denotes that the grip load force coupling deteriorated as a function of coordinative complexity. Together, these data indicate a deficit in the grip-load force constraint due to HD and illustrate the degrading effect of striatal dysfunction on (bi)manual manipulative function. PMID- 11889511 TI - Is human imitation based on a mirror-neurone system? Some behavioural evidence. AB - Recently, a population of neurones was discovered in the monkey's ( Macaca nemestrina) ventrolateral part of the pre-motor cortex (area F5). It is specialised for recognising object-oriented actions, regardless of whether these actions are performed or observed by the monkey. The latter observation led to the term mirror-neurones, and because these cells respond to both observed and executed actions, it seems likely that neurones of that type became co-opted during hominid evolution to serve the imitative behaviours that are so prevalent in our species. There is recent physiological evidence that Broca's area, the human ( Homo sapiens) homologue of monkey's area F5, is involved in the imitation of finger movements. However, concluding that human imitation is based on a mirror-neurone system is premature, because: (1) imitation in monkeys does not reach the same level as in humans or apes and (2) monkeys' mirror-neurones are specialised for object-oriented actions. This specialisation has not yet been demonstrated in adult humans. We investigated the role of objects in human imitation behaviour in a response time experiment. Subjects had to imitate downward movements of an index finger. In one condition, the observed finger touched one of two dots either ipsi- or contralaterally. In the other condition, the very same movements had to be imitated. However, there were no dots on the table. The presence of dots had a decisive influence on error patterns and on response times, but did not influence the movement proper. Dots specifically reduced the onset latency of ipsilateral finger movements and they specifically increased the use of the wrong finger, when contralateral movements were required. In general, results showed that objects also drive human imitation behaviour. Hence, it is very likely that imitation emerged from the mirror neurone system of the common ancestor of monkeys and humans. PMID- 11889512 TI - Induction of persistent changes in the organisation of the human motor cortex. AB - Motor learning must involve changes in the organisation of the brain, and it seems axiomatic that afferent signals generated during repeated motor practice contribute to this. In this study, motor-point stimulation of the first dorsal interosseous (FDI) muscle was paired with transcranial magnetic stimulation of the human motor cortex on three successive days to determine whether repeated stimulation sessions result in enduring reorganisation of the motor cortex. This repeated "dual" stimulation induced significant changes in the excitability of the motor cortex together with expansion of the area of scalp from which these responses were elicited. The expansion in muscle representation was accompanied by large movements in the centre of gravity (CoG), suggesting a true reorganisation of the underlying cortical representational zone. The changes persisted for at least 2 days following the last stimulation session. It is concluded that repeated dual stimulation is capable of inducing long-lasting reorganisation within the motor cortex. These changes may be similar in nature to those seen in the motor cortex during motor learning. Moreover, these observations suggest that it may be possible to induce the motor cortex of patients who have suffered strokes to reorganise in a way that improves the voluntary control of the weakened muscles. PMID- 11889513 TI - The perception of body orientation after neck-proprioceptive stimulation. Effects of time and of visual cueing. AB - Different sensory systems (e.g. proprioception and vision) have a combined influence on the perception of body orientation, but the timescale over which they can be integrated remains unknown. Here we examined how visual information and neck proprioception interact in perception of the "subjective straight ahead" (SSA), as a function of time since initial stimulation. In complete darkness, healthy subjects directed a laser spot to the point felt subjectively to be exactly straight ahead of the trunk. As previously observed, left neck muscle vibration led to a disparity between subjective perception and objective position of the body midline, with SSA misplaced to the left. We found that this displacement was sustained throughout 28 min of continuous proprioceptive stimulation, provided there was no visual input. Moreover, prolonged vibration of neck muscles leads to a continuing disparity between subjective and objective body orientation even after offset of the vibration; the longer the preceding vibration, the more persistent the illusory deviation of body orientation. To examine the role of vision, one group of subjects fixated a central visual target at the start of each block of continuous neck vibration, with SSA then measured at successive intervals in darkness. The illusory deviation of SSA was eliminated whenever visual input was provided, but returned as a linear function of time when visual information was eliminated. These results reveal: the persistent effects of neck proprioception on the SSA, both during and after vibration; the influence of vision; and integration between incoming proprioceptive information and working memory traces of visual information. PMID- 11889514 TI - Visual processing in the ketamine-anesthetized monkey. Optokinetic and blood oxygenation level-dependent responses. AB - We used optokinetic responses and functional magnetic resonance imaging (fMRI) to examine visual processing in monkeys whose conscious state was modulated by low doses (1-2 mg/kg) of the dissociative anesthetic ketamine. We found that, despite the animal's dissociated state and despite specific influences of ketamine on the oculomotor system, optokinetic nystagmus (OKN) could be reliably elicited with large, moving visual patterns. Responses were horizontally bidirectional for monocular stimulation, indicating that ketamine did not eliminate cortical processing of the motion stimulus. Also, results from fMRI directly demonstrated that the cortical blood oxygenation level-dependent (BOLD) response to visual patterns was preserved at the same ketamine doses used to elicit OKN. Finally, in the ketamine-anesthetized state, perceptually bistable motion stimuli produced patterns of spontaneously alternating OKN that normally would be tightly coupled to perceptual changes. These results, taken together, demonstrate that after ketamine administration cortical circuits continue to processes visual patterns in a dose-dependent manner despite the animal's behavioral dissociation. While perceptual experience is difficult to evaluate under these conditions, oculomotor patterns revealed that the brain not only registers but also acts upon its sensory input, employing it to drive a sensorimotor loop and even responding to a sensory conflict by engaging in spontaneous perception-related state changes. The ketamine-anesthetized monkey preparation thereby offers a safe and viable paradigm for the behavioral and electrophysiological investigation of issues related to conscious perception and anesthesia, as well as neural mechanisms of basic sensory processing. PMID- 11889515 TI - A quantitative analysis of the correlations between eye movements and neural activity in the pretectum. AB - The study of the saccadic system has focused mainly on neurons active before the beginning of saccades, in order to determine their contribution in movement planning and execution. However, most oculomotor structures contain also neurons whose activity starts only after the onset of saccades, the maximum of their activity sometimes occurring near saccade end. Their characteristics are still largely unknown. We investigated pretectal neurons with saccade-related activity in the alert cat during eye movements towards a moving target. They emitted a high-frequency burst of action potentials after the onset of saccades, irrespective of their direction, and will be referred to as "pretectal saccade related neurons". The delay between saccade onset and cell activity varied from 17 to 66 ms on average. We found that burst parameters were correlated with the parameters of saccades; the peak eye velocity was correlated with the peak of the spike density function, the saccade amplitude with the number of spikes in the burst, and burst duration increased with saccade duration. The activity of six pretectal saccade-related neurons was studied during smooth pursuit at different velocities. A correlation was found between smooth pursuit velocity and mean firing rate. A minority of these neurons (2/6) were also visually responsive. Their visual activity was proportional to the difference between eye and target velocity during smooth pursuit (retinal slip). These results indicate that the activity of pretectal saccade-related neurons is correlated with the characteristics of eye movements. This finding is in agreement with the known anatomical projections from premotor regions of the saccadic system to the pretectum. PMID- 11889516 TI - Segment difficulty in two-stroke movements in patients with Parkinson's disease. AB - In the horizontal plane on a digitizer tablet, subjects made an elbow-extension, two-stroke movement away from the trunk to a first target and then on to a second target. If the two segments of the movement were executed in an integrative manner, the accuracy constraint on the first segment should have produced changes in kinematic features not only of that segment but also of the second segment. Two-stroke movements of ten Parkinson's disease (PD) patients and ten controls were studied to examine whether a high-accuracy constraint on the first segment influences the performance of the second, when the second target has either a high- or low-accuracy requirement. When the accuracy requirement of the second segment was low, both PD patients and controls showed that changing the first target size from large to small influenced the performance of not only the first segment but also the second segment. For the first segment, movement time, acceleration time, and deceleration time increased when moving to the small first target as compared to the large first target. The peak velocity and peak acceleration also decreased as the first target size decreased. For the second segment, similar patterns of kinematic changes in relation to the first segment were observed in all of these parameters. When the accuracy requirement of the second segment was high, the controls showed similar changes in the first and second segments in relation to the change of first target sizes. In contrast, the PD patients showed that the target size that defined the first movement mainly influenced the performance of that segment. Among kinematic parameters tested for the second segment, only acceleration time increased as the first target size decreased. Other parameters in general did not change, regardless of whether movement of the first segment was made to the small or large target. These results indicate that the two-stroke movements of PD patients showed little evidence that they were planned and organized in an integrative manner when there was a high-accuracy constraint imposed on the second segment. On the other hand, control subjects performed two-stroke movements in a manner that suggested the two segments were planned and organized together regardless of an accuracy constraint imposed. PMID- 11889519 TI - Future directions of congenital heart disease imaging. PMID- 11889520 TI - A simple method for evaluating abnormal lengthening of the QT interval during the face immersion test. AB - The slope of the relation between the unadjusted QT interval and heart rate during the face immersion test has been reported to be useful as an index for predicting an abnormal lengthening of the QT interval for children with nonfamilial long QT syndrome. Our goals were to determine whether we can replace the slope of the QT/heart rate relation calculated from all data with that calculated from fewer data and to determine whether we can replace the slope with the corrected QT value by heart rate (QTc value) at the minimum heart rate. We studied 19 children with a prolonged QT interval and 54 control children by using statistical analysis. The slope calculated from the selected data points (at least four) was in agreement with the slope calculated from all data, and the relationship between the slope and the QTc value at the minimum heart rate showed a high correlation. It was determined that we can replace the slope calculated from all data with that calculated from at least four data points and replace the slope with the QTc value at the minimum heart rate as an index for predicting an abnormal lengthening of the QT interval. PMID- 11889521 TI - Determination of the pressure gradient in children with coarctation of the aorta by low-field magnetic resonance imaging. AB - During the past few years magnetic resonance imaging (MRI) has gained increasing importance in the noninvasive examination of congenital heart defects. Practically all existing examinations have been carried out with a magnetic field strength exceeding 1 (T high-field MRI). Flow quantification is considered to be an advanced MRI application and, in the past, has been available for high-field systems only. Therefore until recently, functional examinations such as MRI tomographic flow quantification were reported exclusively for high-field MRI units. From December 1998 to December 1999, nine patients (five girls and four boys, mean age 130 plus minus 3.6 months, range 62-185 months) were investigated by means of MRI after a previous surgical repair or interventional balloon dilatation of a coarctation of the aorta (mean postinterventional time period 23 plus minus 0.4 months). The examination was carried out without sedation in an open low-field unit (Siemens Magnetom Open 0.2 T). Cardiac-triggered spin-echo sequences were used with a 3-mm to 7-mm slice thickness in an axial and a double oblique plane. The measurement of the immediate poststenotic flow velocity was done by flow-sensitive sequences developed for the study (phase-sensitive flow measurement sequences: TE, 6-12 msec; TR, 50 msec; flip angle, 60 degrees; Vmax, 200-1200 cm/sec; two acquisitions). All patients were examined on the same day with comparative echocardiographic procedures. In all cases, an excellent anatomical evaluation of the aortic arch was possible. The diameters of the residual stenosis were measured by MRI and correlated well r = 0.95; p ? 0.001; mean difference 0.44 +/- 2.47 mm) with echocardiographic results. No wall damage was observed in any of the cases studied. The pressure gradient of the stenosis calculated from the flow sequence was between 17 and 50 mmHg and corresponded well (r = 0.93; p = 0.001; mean difference 0.67 +/- 11 mmHg) with the results obtained from echocardiography. The study demonstrates that examination of the aortic arch is possible in a low-field MRI system, with its significant advantages (lower patient discomfort and more cost-effective examination). In addition, a quantitative flow measurement in low-field MRI was realized for the first time. Low-field MRI therefore seems to be a good, noninvasive method for examining patients with a poor echocardiographic representation of the aortic arch. PMID- 11889522 TI - Plasma levels of nitrate in congenital heart disease: comparison with healthy children. AB - Nitric oxide (NO) is an endothelium- derived relaxing factor, and plasma nitrate is the stable end product of NO production. The aim of this study was to investigate the change in levels of plasma nitrate according to age and to elucidate the effect of pulmonary hypertension (PH) associated with congenital heart disease on NO production. We measured plasma levels of nitrate in 48 healthy children aged 5 days to 12 years to establish the normal range. Forty-six preoperative patients aged 4 months to 12 years with congenital heart disease were studied by cardiac catheterization. Plasma nitrate in healthy children decreased with age, from 1 month to 1 year, and then remained almost constant until the age of 12 years. Plasma nitrate was significantly increased in 22 preoperative patients with PH (mean pulmonary arterial pressure >25 mmHg) compared with age-matched normal controls: (mean 56.9 vs 33.5 micromol/L, p<0.05) and was significantly correlated with pulmonary to systemic pressure ratio (r= 0.83, p < 0.0001). There was no significant difference between plasma nitrate levels in 24 preoperative patients without PH and those in the age-matched normal control (mean 25.6 vs 24.9 micromol/L). In 10 patients with preoperative PH who were examined before and after surgery, plasma nitrate levels remained high in the cases with residual PH but decreased to the normal range in the cases without residual PH. Plasma nitrate level is useful for evaluating PH both before and after operation in patients more than 4 months of age, and it is important to note differences in normal plasma nitrate levels according to age. PMID- 11889523 TI - Outcome of 121 patients with congenitally corrected transposition of the great arteries. AB - Congenitally corrected transposition of the great arteries (ccTGA) is a rare disorder with reduced survival that is influenced by the presence of associated anomalies, tricuspid regurgitation (TR), and right ventricular (RV) function. The double switch procedure has been proposed as an aggressive surgical approach in selected patients. We sought to review our experience with conventional repair to determine if a change in surgical strategy was warranted. Clinical records of 121 patients with ccTGA and two adequate-sized ventricles were retrospectively reviewed. Median length of follow-up was 9.3 years; 5-, 10-, and 20-year survival rates were 92%, 91%, and 75%, respectively. Surgery was performed in 86 patients, including conventional biventricular repair in 47 patients. Risk factors for mortality by univariate analysis included age at biventricular repair (p = 0.04), complete atrioventricular (AV) canal defect (p = 0.02), dextrocardia (p = 0.05), moderate or severe TR (p = 0.05), and poor RV function (p = 0.001). By multivariate analysis, complete AV canal defect (p = 0.006) and poor RV function (p = 0.002) remained significant as risk factors for mortality. Risk factors for the development of significant TR included conventional biventricular repair (p = 0.03) and complete AV block (p = 0.04). Risk factors for progressive RV dysfunction included conventional biventricular repair (p = 0.02), complete AV block (p = 0.001), and moderate or severe TR (p < 0.001). This is the largest nonselected cohort of patients with ccTGA followed at a single center. Our results confirm that significant TR and poor RV function are risk factors for poor outcome and provide convincing evidence that patients undergoing conventional biventricular repair are at higher risk for deterioration of tricuspid valve and right ventricular function compared to palliated or unoperated patients. We support a move toward an alternative surgical approach (double switch procedure) in carefully selected patients. PMID- 11889524 TI - The influence of a restrictive atrial septal defect on pulmonary vascular morphology in patients with hypoplastic left heart syndrome. AB - Hypoplastic left heart syndrome (HLHS) with a restrictive atrial septal defect (ASD) is a form of congenital heart disease with considerable morbidity and mortality. This morphologic analysis assesses the pulmonary vasculature in this patient population. Pulmonary arteries, the persistence of high-resistance fetal arterioles, pulmonary veins, and lymphatics from multiple lung sections from each of five patients with HLHS and a restrictive ASD were compared to those of five patients with HLHS and nonrestrictive ASD. Lung sections from each patient were qualitatively graded in severity of pathology from 0 to 3 for each of the structures described previously, with the pathologist blinded to the status of the ASD. Patients with a restrictive ASD exhibited more significant pulmonary venous thickening and lymphatic dilatation (p = 0.02), with a tendency toward persistence of high-resistance fetal vessels (p = 0.2), compared to patients with a nonrestrictive ASD. These findings imply that patients with HLHS and a restrictive ASD possess pulmonary vascular abnormalities that place them at higher risk for the current surgical interventions available compared to patients with a nonrestrictive ASD. PMID- 11889525 TI - Around PediHeart: plastic bronchitis. PMID- 11889526 TI - Analysis of atrioventricular plane movements by Doppler tissue imaging and M-mode in children with atrial septal defects before and after surgical and device closure. AB - Our objective was to compare the effects of surgical and device closure of atrial septal defects (ASDs) on atrioventricular plane function. In healthy individuals, both short- and long-axis motion contribute to ventricular pump function. Short axis function (i.e., the amplitude and velocity of atrioventricular plane movements) may be evaluated by M-mode and Doppler tissue imaging. The study group consisted of 19 children with ASD of the secundum type before and after surgical (n = 12) or device (n = 7) closure and 10 healthy controls. Surgical and device closures were uncomplicated and all defects were completely closed. Registrations of atrioventricular plane systolic and diastolic amplitude and velocity were made from the tricuspid and the mitral annulus and from the septum in the apical four chamber view. Comparisons were made between examinations before and after closure, between the two subgroups of patients treated by surgical and device closure, respectively, and between the patient group and the control groups. Before ASD closure, all measurements were normal or near normal. After surgery, systolic amplitudes and velocities of the tricuspid annulus and in the septum decreased significantly, whereas no changes were seen in the device group. Less marked changes were seen in diastolic measurements. However, in the surgical group significant decrease to subnormal values were found in the tricuspid annulus and in the septum, which may indicate a decreased diastolic function postoperatively. Mitral valve annulus amplitude and velocity were not affected by the treatment. Atrioventricular function is normal in children with right ventricular volume overload. The decrease to subnormal values after open-heart surgery is not seen after device closure of the ASD, indicating that surgery affects right ventricular function. PMID- 11889527 TI - Plasma levels of natriuretic peptide and echocardiographic parameters in patients with Duchenne's progressive muscular dystrophy. AB - We investigated the relationship between plasma atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) levels and systolic and diastolic cardiac function, determined by echocardiography, in 63 patients with Duchenne's progressive muscular dystrophy (DMD) (age range 8-21 years). The relationship between shortening fraction of the left ventricle and ANP and BNP levels was curvilinear rather than linear: When the shortening fraction was >15%, increases in ANP and BNP levels were minimal. However, if the shortening fraction was <15%, both natriuretic peptide levels increased dramatically. Stepwise regression analysis revealed that only the deceleration time of the early diastolic filling wave predicted plasma BNP concentration among various diastolic echocardiographic parameters determined by mitral flow. Three patients died of cardiac dysfunction during a 2-year follow-up period. These patients had a severely decreased deceleration time (<65% of normal) in association with increases in both natriuretic peptide levels. In conclusion, plasma ANP and BNP levels are not sensitive markers for the early detection of cardiac systolic dysfunction in patients with DMD. However, in patients with systolic dysfunction, an increase in the concentrations of these peptides, associated with a decrease in the deceleration time of early diastolic filling, suggests poor prognosis. PMID- 11889528 TI - Right ventricular performance in hypotensive preterm neonates treated with dopamine. AB - Systemic hypotension with left ventricular dysfunction is a common complication of neonatal respiratory distress syndrome and is often treated with inotropic agents. Although pulmonary hypertension with elevated pulmonary vascular resistance is also an important pathophysiological finding in respiratory distress syndrome, the effect of inotropes on the right ventricle has not been studied. The aim of this study was to assess changes in right ventricular dimensions and function with inotropic therapy in hypotensive preterm infants. Hypotensive neonates with respiratory distress syndrome were studied before and 1 hour after the initiation of a dopamine infusion. Right ventricular performance was assessed by two-dimensional echocardiography using the ellipsoid approximation method. Eight hypotensive neonates were recruited with a median (interquartile range) gestation of 27 weeks (26 to 27 weeks). Right ventricular end systolic volume decreased significantly from a median (interquartile range) of 1.06 ml/kg (0.81-1.50 ml/kg) to 0.73 ml/kg (0.51-0.99 ml/kg) (p < 0.01) 1 hour following dopamine therapy. Right ventricular end diastolic volume did not change significantly. Right ventricular ejection fraction increased significantly from 0.36 (0.29-0.46) to 0.51 (0.43-0.53) ( p < 0.01). There was a trend toward an increase in right ventricular output from 90 ml/kg/min (67-115 ml/kg/min) to 112 ml/kg/min-143 ml/kg/min) (p=0.07). Dopamine increases right ventricular ejection fraction through a reduction in right ventricular end systolic volume. PMID- 11889529 TI - Effect of vessel size on the flow efficiency of the total cavopulmonary connection: in vitro studies. AB - The total cavopulmonary connection (TCPC) creates a passive system of blood flow into the pulmonary circulation. We hypothesize that the efficiency differences found in models with superior vena cava-inferior vena cava (SVC-IVC) offsets is dependent on vessel size, with flow efficiency improving with larger size vessels. Two sets of in vitro TCPC models (TCPC-3 and TCPC-15) were constructed corresponding to average vessel diameters of 3- and 15-year-old patients. The model with full SVC-IVC offset was the most efficient in TCPC-3 models. There was no significant difference between geometric arrangements in TCPC-15 models; however, the average efficiencies were significantly higher. Among the models representing vessel sizes of the younger age group, the model with the full diameter SVC-IVC offset was the most efficient. The models representing vessel sizes of the older age group showed marked improvement in efficiencies throughout without obvious differences between various geometric arrangements. This suggests that geometric considerations in TCPC surgical techniques may be of lower than expected significance over the life span of a patient. More important, after balancing the effects of improved flow efficiency with larger vessels against the effects of chronic volume overload, the trend of performing the Fontan surgery on increasingly younger patients may need to be reconsidered after further studies. PMID- 11889530 TI - False aneurysm following modified Blalock-Taussig shunt. AB - A 3 1/2-year-old female child presented with massive hemoptysis 5 months after a modified Blalock-Taussig (BT) shunt for double-outlet right ventricle with pulmonary stenosis. Computerized tomographic scan and angiography showed a false aneurysm of the subclavian artery at the insertion of the shunt. Successful surgical management is discussed. PMID- 11889531 TI - Around PediHeart: trisomy 18, an ethical dilemma. PMID- 11889532 TI - Reversal of pulmonary hypertension associated with plexiform lesions in congenital heart disease: a case report. AB - We describe a 2-year-old child with severe pulmonary hypertension due to a patent ductus arteriosus (PDA) with plexiform lesions on lung biopsy. Despite high basal pulmonary vascular resistance with minimal responsiveness to inhaled nitric oxide and other vasodilators, and advanced plexogenic arteriopathy on lung biopsy, her pulmonary hypertension completely resolved after PDA ligation and during 8 years of follow-up. PMID- 11889533 TI - A calculus of unnecessary echoes: application of management principles to health care. AB - We studied the clinical utility of echocardiography in children and applied principles of business management to draw conclusions that are applicable to health care in general. A significant number (13% in this series) of expensive medical diagnostic tests could be avoided without harm to patients. Cost reduction in medicine is possible in many situations without compromising quality of care. Care pathways (i.e., practice guidelines or clinical algorithms) provide one useful modality. However, for the safety of patients, all cost reduction methods must start with practicing physicians (or involve them at conceptualization) and an escape clause must be available to the treating physician for the atypical patient. The analytic approach used--concurrent assessment of percentage cost, charge, and payor--is applicable to all components of the health care value chain. The use of "percentage of charges" as an indicator of collection effectiveness is unrealistic and should be changed to "percentage potential reimbursement" because health care is effectively a fixed reimbursement industry rather than a system subject to standard microeconomic (supply and demand) forces. The current reimbursement structure provides conflicting incentives both to health care institutions and to providers, creating an insurmountable barrier to any effective incentive system. Colloquy between practicing physicians and experts in operations management will stimulate cost reduction and can optimize the delivery of health care. PMID- 11889534 TI - Assessment of the ability of myocardial contrast echocardiography with harmonic power Doppler imaging to identify perfusion abnormalities in patients with Kawasaki disease at rest and during dipyridamole stress. AB - The aim of our study was to assess the ability of myocardial contrast echocardiography (MCE) with harmonic power Doppler imaging (HPDI) to identify perfusion abnormalities in patients with Kawasaki disease at rest and during pharmacological stress imaging with dipyridamole. Results were compared with those of 99mTc-tetrofosmin single-photon emission computed tomography (SPECT) imaging as the clinical reference standard. MCE with HPDI was performed on 20 patients with a history of Kawasaki disease. Images were obtained at baseline and during dipyridamole infusion (0.56 mg x kg(-1)) in the apical two- and four chamber views. Myocardial opacification suitable for the analysis was obtained in all patients. Nine patients with stenotic lesions had a reversible defect after dipyridamole infusion detected by both MCE with HPDI and SPECT, and 3 patients with a history of myocardial infarction had a partially or completely irreversible defect detected by both methods. Three patients with coronary aneurysm without stenotic lesion, 4 patients with regressed coronary aneurysm, and 2 patients with normal coronary artery in acute phase also had normal perfusion at rest and after pharmacological stress by both methods. A 96% concordance (kappa = 0.87) was obtained when comparing the respective segmental perfusion scores using the two methods at baseline, and an 86% concordance (kappa = 0.81) was obtained at postdipyridamole infusion. After combining baseline and postdipyridamole images, each segment was labeled as having normal perfusion, irreversible defects, or reversible defects. Using these classifications, concordance for the two methods was 92% (kappa = 0.87). MCE with HPDI is a safe and feasible method by which to detect asymptomatic ischemia due to severe stenotic lesion, and it may be an important addition to the modalities used to identify patients at risk for myocardial infarction as a complication of Kawasaki disease. PMID- 11889535 TI - Ventriculography using ECG-gated multiple diastolic injection of contrast material in pediatric angiocardiography. AB - In pediatric patients, the lower body weight limits the volume of contrast medium. Conventionally, angiocardiography is carried out with a single large bolus injection of contrast material. Angiocardiograms are used in pediatric patients with complex congenital heart diseases. In patients with complex congenital heart disease, especially with MAPCA, the volume of contrast medium used may be excessive. This would allow further injection to provide additional information. To reduce contrast medium used in the angiocardiogram in pediatric patients, we decided to use electrocardiogram (ECG)-gated multiple diastolic injection (EMDI). Three small boluses were injected during the diastolic phase of three consecutive cardiac cycles using a commercially-available power injector. Seventy-eight ventriculograms (47 left ventriculograms and 31 right ventriculograms) using EMDI were carried out on 53 patients with congenital heart disease. Total contrast medium volume with EMDI ventriculograms (mean [+/- SD] per body weight: 0.72 [+/- 0.25] ml/kg) was significantly smaller than with conventional injection (1.01 [+/- 0.36] ml/kg) (p <0.001). The grades of ventriculograms with EMDI tended to be slightly better than those with conventional injection (statistically not significant, p = 0.478). No short-run type premature ventricular contraction (PVC)s or intramural injection occurred in the ventriculograms with EMDI. PVCs tended to be less frequent in the ventriculograms with EMDI than in those with conventional injection (statistically not significant, p = 0.131). EMDI may be worthwhile in reducing ventricular ectopy when checking ventricular function by angiography, since hemodynamic conditions are less affected by small quantities of contrast medium and only during the filling phase of the ventricles. In conclusion, EMDI may be a useful method for reducing complications of ventriculography in pediatric angiocardiography. PMID- 11889536 TI - Berry syndrome with trisomy 13. AB - We report on 2-day-old neonate with trisomy 13 with coexistent distal aortopulmonary septal defect, aortic origin of the right pulmonary artery, interrupted aortic arch, intact ventricular septum, and a patent ductus arteriosus diagnosed by two-dimensional and color Doppler echocardiography. Review of the literature reveals that this patient is the 24th reported case of Berry syndrome and the first case of this unusual combination of cardiovascular defects associated with trisomy 13 syndrome. PMID- 11889537 TI - Resolution of acquired pulmonary arteriovenous malformations in a patient with total anomalous systemic venous return. AB - An 11-year-old male with total anomalous systemic venous return had surgical repair except for the hepatic venous return, which drained to the left atrium. He developed progressive cyanosis and fatigue and was diagnosed with large pulmonary arteriovenous malformations (PAVMs) during cardiac catheterization with the use of bubble contrast echocardiography. After surgical redirection of hepatic venous flow to the right heart and pulmonary arterial system, resolution of these PAVMs was demonstrated clinically and by contrast echocardiography. This unique case report demonstrates the development of PAVMs with exclusion of hepatic venous return through the pulmonary vascular bed while pulsatile pulmonary blood flow remains intact. It reinforces the likelihood of the absence of an as yet unidentified hepatic vasoactive substance as the source for development of PAVMs. PMID- 11889538 TI - Resolution of electrical storms after discontinuation of ICD therapy in a child with long QT syndrome. AB - We report a case of a child with familial long QT syndrome (Jervell Lange Nielsen) who had multiple electrical storms in the presence of b blocker and implantable cardioverter device (ICD) therapy. Discontinuation of ICD therapy and addition of oral amiodarone to b blockade therapy resulted in freedom from electrical storms. PMID- 11889539 TI - Around PediHeart: pulmonary arteriovenous malformations. PMID- 11889540 TI - Pseudohypertrophic muscular dystrophy with pulmonary valve stenosis: an unusual association. AB - A patient with pulmonary valve stenosis associated with pseudohypertrophic muscular dystrophy is reported. The anomalies were detected by two-dimensional echocardiography and elevated serum enzymes, conformed by right ventriculography and muscles biopsy. The association between these two pathologies is unknown. PMID- 11889541 TI - Congenital coronary arteriovenous fistula presenting with syncope. AB - Previous reports of syncope in patients with coronary arteriovenous fistula (CAVF) have theorized that it occurs secondary to a coronary steal phenomenon. We present a case of syncope in a young woman with a CAVF and no anatomic substrate for coronary steal. PMID- 11889542 TI - Serial stent implantation to relieve in-stent stenosis in obstructed total anomalous pulmonary venous return. AB - A nine-year-old girl with complex cyanotic heart disease associated with supracardiac total anomalous venous return and obstruction at the drainage site of the vertical vein into the left innominate vein had stent placement at the age of seven, with immediate increase of oxygen saturation and improvement of physical activity. Significant in-stent stenosis occurred that was successfully treated by concentrical placement of a second stent. This case report demonstrates the transluminal approach to be effective in the treatment of stenoses in congenital heart defects that are not eligible for corrective surgery. PMID- 11889543 TI - Exercise performance in tetralogy of Fallot: the impact of primary complete repair in infancy. AB - Primary complete repair (PCR) of tetralogy of Fallot (TOF) is now routinely performed in infancy. Although operative results are excellent, the impact on exercise performance is incompletely understood. We reviewed data of all children with TOF who underwent PCR at our institution and had subsequent maximal cycle ergometer exercise testing between January 1995 and December 2000. Of the 193 patients with TOF who underwent PCR, 57 (30%) underwent exercise testing; maximal tests were available for 50 of 57 (88%). Exercise performance of subjects who underwent PCR at <1 year of age was compared to that of those who underwent repair at >l year of age. The median age at PCR was 10.9 months; 28 subjects (56%) underwent PCR in infancy (<1 year). A transannular incision was employed in the repair in 41 subjects (82%). The mean age at exercise testing was 12.5 +/- 3.2 years. The mean maximal VO2 was 94.9 +/- 18.8% predicted and the mean maximal work rate was 98.0 +/- 20.8% predicted. In multivariate analysis PCR in infancy (age <1 year) was not associated with maximal VO2, peak work rate, peak heart rate, or arrhythmias. Only older age at testing and male gender were significantly associated with higher maximal VO2 (p = 0.005 and p = 0.002, respectively). Intermediate-term exercise performance in subjects who undergo PCR of TOF in early childhood is near normal. Performing PCR in the first year of life does not impact subsequent exercise performance. PMID- 11889544 TI - Single origin of right and left pulmonary artery branches from ascending aorta with nonbranching main pulmonary artery: relevance to a new understanding of truncus arteriosus. AB - We report the third known case of origin of the right and left pulmonary artery branches from the ascending aorta via a short common pulmonary artery. A large unbranching main pulmonary artery opened through a patent ductus arteriosus into the descending thoracic aorta. Preductal coarctation of the aorta and multiple congenital anomalies were also present. This rare cardiovascular malformation facilitates a new anatomic and developmental understanding of truncus arteriosus. PMID- 11889545 TI - Coil embolization of a neonatal pulmonary arteriovenous malformation. AB - Pulmonary arteriovenous malformation (PAVM), as a part of Osler-Weber-Rendu Syndrome, in the neonate, is a rare hereditary vascular malformation. Large intrapulmonary right-to-left shunting, causing hypoxaemia and cyanosis, can be a life-threatening condition. Repeated transcatheter coil embolization procedures proved to be a favorable strategy to improve systemic arterial oxygen saturation, with a good outcome in a newborn child. While the radiation dose was high, the use of this amount of radiation was felt to be justified and its effects considered tolerable in the treatment of this patient's serious malformation. PMID- 11889547 TI - Truncus arteriosus with a very small ventricular septal defect diagnosed by echocardiography. PMID- 11889546 TI - Idiopathic severe pulmonary hypertension in an infant with pulmonary infection. AB - We report a 5-month-old infant who showed typical echocardiographic findings of primary pulmonary hypertension without the typical histopathological findings and who recovered from severe pulmonary hypertension. Histopathological findings revealed mild thickening of small pulmonary arteries and activated macrophages in the lung. Some cases with idiopathic severe pulmonary hypertension in infants are associated with pulmonary infection. PMID- 11889548 TI - Double aortic arch with interruption proximal to the right carotid artery, bilateral patent ductus arteriosi, and complex congenital heart disease. PMID- 11889549 TI - Consideration of the respiratory phase in diagnosing cardiac enlargement in the chest X-ray of children. PMID- 11889550 TI - Protein-losing enteropathy. PMID- 11889553 TI - Molecular characterization of a mouse cDNA encoding Dicer, a ribonuclease III ortholog involved in RNA interference. AB - Members of the ribonuclease III superfamily of double-stranded(ds)-RNA-specific endoribonucleases participate in diverse cellular RNA maturation and degradation pathways. A recently identified eukaryotic RNase III family member, named "Dicer", functions in the RNA interference (RNAi) pathway by producing 21--23 bp dsRNAs which target the selective destruction of homologous RNAs. RNAi is operative in animals, plants, and fungi, where it is proposed to inhibit viral reproduction and retroposon movement, as well as to participate in developmental pathways. RNAi functions in mammalian cells, including mouse oocytes and embryos. This article reports the cDNA sequence characterization and expression analysis of the mouse Dicer ortholog. On the basis of the cDNA sequence, the Dicer polypeptide is 1906 amino acids and has a predicted molecular mass of 215 kDa. Mouse Dicer contains a DExH/DEAH helicase motif; a PAZ domain; a tandem repeat of RNase III catalytic domain sequences; and a dsRNA-binding motif. The Dicer gene maps to a single locus on the distal portion of mouse Chromosome (Chr) 12. The Dicer transcript is expressed from the embryonic through adult stages of development. The Dicer transcript is also present in a wide variety of adult mouse organs. The highly conserved set of functional domains and the occurrence of a single-copy gene strongly indicate that the encoded protein is the RNase III ortholog responsible for dsRNA processing in the RNAi pathway. PMID- 11889554 TI - Alternative non-coding splice variants of Nespas, an imprinted gene antisense to Nesp in the Gnas imprinting cluster. AB - The Gnas locus on mouse Chr 2 represents a unique cluster of overlapping imprinted genes. Three of these in the order Nesp--Gnasxl--Gnas are transcribed in the sense direction with Nesp having maternal-specific expression, Gnasxl having paternal expression, and Gnas as being biallelically expressed in most tissues. A fourth imprinted gene, Nespas, is paternally expressed, lies antisense to Nesp, and expresses an unspliced transcript. Large unspliced antisense transcripts are emerging as a feature of imprinted gene clusters, and such non coding RNAs may have a cis-regulatory function. Here we show that, in addition to an unspliced form of Nepas, we can detect five alternatively spliced forms of Nespas up to 1.4 kb in length that are non-coding. The splice variants are paternally expressed; they start approximately 2 kb upstream of Gnasxl in a region of maternal methylation and end 2.5 kb beyond the ATG of Nesp. These variants do not correspond to exons of the human antisense transcript although they start in the same region; the Nespas transcript, like its human counterpart, is spliced in various alternative patterns. The identification of a set of small spliced imprinted transcripts in the human and now in the mouse suggests that these antisense transcripts are functionally important. PMID- 11889555 TI - Assignment of 280 swine genomic inserts including 31 microsatellites from BAC clones to the swine RH map (IMpRH map). AB - A precise genetic map containing anonymous markers and genes is indispensable for the efficient selection of candidate gene(s) responsible for quantitative trait loci (QTL) traits. For this purpose, a first version of a radiation hybrid cell (RH) map has been constructed by using the INRA-University of Minnesota RH panel for 757 markers (IMpRH) (Hawken et al. 1999, Mamm. Genome 10: 824--830). In this study, 280 swine genomic fragments in BAC clones were assigned to the IMpRH map; 255 BAC clones were successfully linked to first-generation linkage groups (LOD > 4.8). The remaining 25 clones could not be mapped, because their lod-scores to the closest markers in the first generation map were less than 4.8. In addition, 16 BAC clones, mapped to swine Chromosome (Chr) 1 by IMpRH mapping, were subjected to isolation of microsatellites (MSs). Thirty-one MSs were isolated from 15 BAC clones, and 24 of 31 (77%) MSs derived from 14 clones were found to be polymorphic. We also mapped both termini of 12 BAC clones to the IMpRH map, in order to measure resolution of the IMpRH map; the resolution was found to range from 8 kb/centiRay to more than 126 kb/centiRay depending on the region. PMID- 11889556 TI - Construction of a 5000(rad) whole-genome radiation hybrid panel in the horse and generation of a comprehensive and comparative map for ECA11. AB - A 5000(rad) whole-genome radiation hybrid (RH) panel was created for the horse. The usefulness of the panel for generating physically ordered maps of individual equine chromosomes was tested by typing 24 markers on horse Chromosome 11 (ECA11). The overall retention of markers on this chromosome was 43.6%. Almost complete retention of two of the typed markers--- CA062 and AHT44---clearly indicated the location of thymidine kinase gene on the short arm of ECA11. Seven of the typed markers were FISH mapped to align the RH and cytogenetic maps. With the RH-MAPPER approach, a physically ordered map comprising four linkage groups and incorporating all the markers was obtained. The study provides the first comprehensive map for a horse chromosome that integrates all available mapping data and adds new information that spans the entire length of the equine chromosome. The map clearly underlines the resolving power and utility of the panel and emphasizes the need to have uniformly distributed cytogenetic markers for appropriate alignment of RH map with the chromosome. A comparative status of the ECA11 map in relation to the corresponding human/mouse chromosome is presented. PMID- 11889558 TI - Characterization of a novel gene adjacent to PAX6, revealing synteny conservation with functional significance. AB - The human eye anomaly aniridia is normally caused by intragenic mutations of PAX6. Several cases of aniridia are, however, associated with chromosomal rearrangements that leave the PAX6 gene intact. We have identified and characterized a novel gene, PAXNEB (C11orf19), downstream (telomeric) of PAX6. Sequence analysis, including interspecies comparisons, show this gene to consist of 10 exons, with an unusually large final intron spanning 134 kb in human and 18 kb in Fugu. This intron is disrupted by each chromosomal rearrangement. The 2-kb PAXNEB transcript, encoding a 424-amino acid protein, is expressed in all cell lines tested. The homologous mouse cDNA is broadly expressed in mouse embryos. PAXNEB is highly conserved from mammals to fish, with some regions of the protein showing conservation to invertebrates, yeast, and plants. The possible role of PAXNEB in aniridia was assessed. Using a transgenic mouse model, we show that the aniridia phenotype of the chromosomal rearrangement cases is not due to the heterozygous loss of PAXNEB function. PMID- 11889557 TI - Molecular characterization and mapping of ATOH7, a human atonal homolog with a predicted role in retinal ganglion cell development. AB - The human ATOH7 gene encodes a basic helix-loop-helix (bHLH) transcription factor that is highly similar to Drosophila Atonal within the conserved bHLH domain. The ATOH7 coding region is contained within a single exon. We mapped ATOH7 to Chromosome (Chr) 10q21.3-22.1, a region syntenic to the segment of mouse Chr 10 where Atoh7 (formerly Math5) is located. The evolutionary relationship between ATOH7 and other atonal homologs was investigated using parsimony analysis. A direct comparison of ATH5/7 and ATH1 protein subgroups to Atonal also revealed a nonrandom distribution of amino acid changes across the bHLH domain, which may be related to their separate visual and proprioceptive sensory functions. Among bHLH genes, ATOH7 is most closely related to Atoh7. This sequence conservation extends significantly beyond the coding region. We define blocks of strong homology in flanking human and mouse genomic DNA, which are likely to include cis regulatory elements. Because targeted deletion of Atoh7 causes optic nerve agenesis in mice, we propose ATOH7 as a candidate for human optic nerve aplasia and related clinical syndromes. PMID- 11889559 TI - Molecular basis of the Cd36 chromosomal deletion underlying SHR defects in insulin action and fatty acid metabolism. AB - The human insulin resistance syndromes---type 2 diabetes, obesity, combined hyperlipidemia, and essential hypertension---are genetically complex disorders whose molecular basis is largely unknown. The spontaneously hypertensive rate (SHR) is a model of these human syndromes. In the SHR/NCrlBR strain, a chromosomal deletion event that occurred at the Cd36 locus during the evolution of this SHR strain has been proposed as a cause of defective insulin action and fatty acid metabolism. In this study, three copies of the Cd36 gene, one transcribed copy and two pseudogenes, were identified in normal rat strains, but only a single gene in SHR/NCrlBR. Analysis of SHR genomic sequence localized the chromosomal deletion event between intron 4 of the normally transcribed copy of the gene and intron 4 of the second pseudogene. The deletion led to the creation of a single chimeric Cd36 gene in SHR/NCrlBR. The boundaries of the recombination/deletion junction identified within intron 4 were surrounded by long interspersed nuclear elements (LINEs) and DNA topoisomerase I recognition sequences. An 8-bp deletion at the intron 14/exon 15 boundary of the second pseudogene abolishes the putative splice acceptor site and is the cause of an aberrant 3' UTR previously observed in SHR/NCrlBR. We conclude that in SHR/NCrlBR, the complex trait of insulin resistance and defective fatty acid metabolism is caused by Cd36 deficiency, resulting from a chromosomal deletion caused by unequal recombination. This demonstrates that chromosomal deletions caused by unequal recombination can be a cause of quantitative or complex mammalian phenotypes. PMID- 11889560 TI - Fluctuation in fertility phenotypes of the heterozygous ( Om/+) mice owing to background genes. PMID- 11889561 TI - CHD5 defines a new subfamily of chromodomain-SWI2/SNF2-like helicases. PMID- 11889562 TI - Orthologs of seven genes (AKT1, CDC42BPB, DIO3, EIF5, JAG2, KLC, NDUFB1) from human chromosome 14q32 map to distal chicken chromosome 5. PMID- 11889563 TI - Nitric oxide enhances Ca(2+)-dependent K(+) channel activity in rat carotid body cells. AB - The nitric oxide (NO) donor S-nitroso-acetylpenicillamine (SNAP) enhanced Ca(2+) dependent K(+) channel activity in rat carotid body chemoreceptor cells. Ca(2+) dependent K(+) channel activity was enhanced by SNAP in 38% (whole-cell configuration) and 67% (cell-attached mode) of the cells tested and was not affected by intracellular Ca(2+) chelation with BAPTA-AM. Enhancement of Ca(2+) dependent K(+) channel activity by SNAP was blocked by the cGMP-dependent protein kinase G inhibitor 8-[(4-chlorophenyl)thio]-guanosine 3',5'-cyclic monophosphothioate Rp diastereomer (Rp-8-pCPT-cGMPS). NO thus enhances Ca(2+) dependent K(+) channel activity through cGMP-dependent protein kinase G. The NO mediated increase in Ca(2+)-dependent K(+) channel activity is likely to alter the function of carotid body chemoreceptor cells and could explain the decreased chemosensitivity of the carotid body in response to NO released from efferent nerves or vascular endothelial cells. PMID- 11889564 TI - Expression and regulation of connexins in cultured ventricular myocytes isolated from adult rat hearts. AB - Gap junctions were assayed during re-differentiation of adult rat cardiomyocytes in long-term culture to gain insight into the processes of remodeling. Double immunostaining allowed the localization of connexins Cx40, Cx43, and Cx45 between myocytes and demonstrated co-expression and co-localization in individual cells and gap junction plaques, respectively. Immunoblots showed differential time dependent changes in connexin expression and phosphorylation. The total amount of connexins and the ratio of phosphorylated/non-phosphorylated isoforms gradually increased during the re-establishment of intercellular communication. Dual voltage-clamp studies showed the involvement of several types of gap junction channels. Multichannel currents yielded diverse spectra of g(j,inst)=f( V(j)) and g(j,ss)=f( V(j)) relationships ( g(j,inst): instantaneous gap junction conductance; g(j,ss): conductance at steady state; V(j): transjunctional voltage), indicative of homotypic and heterotypic channels. Single-channel currents revealed two prominent conductances reflecting gamma(j,main) and gamma(j,residual). The histograms of gamma(j,main) showed four discrete peaks (41 44, 59-61, 70-76, and 100-107 pS) attributable to a combination of Cx45-Cx45, Cx40-Cx45 and Cx43-Cx45 channels (1st peak), Cx43-Cx43 and Cx40-Cx43 channels (2nd peak), Cx43-Cx43 channels (3rd peak) and Cx40-Cx40 and Cx40-Cx43 channels (4th peak). However, the presence of heteromeric channels cannot be excluded. The data are consistent with an up-regulation of Cx45 and Cx43 during re differentiation. PMID- 11889565 TI - Early effects of exercise training on on- and off-kinetics in 50-year-old subjects. AB - We tested the hypothesis that, in healthy middle-aged subjects ( n=11, age 51.0 +/- 3.0 years, x +/- SD), the effects of exercise training on pulmonary O(2) uptake (VO(2)) on- and off-kinetics would appear earlier than those on peak. The subjects underwent a standard training program (combined endurance and resistance training) in a health club, and were evaluated before training ("time 0", T0), and after 7 (T7), 15 (T15), 30 (T30), 60 (T60) and 90 (T90) days of training. Breath-by-breath pulmonary O(2) uptake (VO(2)), heart rate (HR), systolic (SBP) and diastolic blood pressure, and capillary blood lactate concentration ([La](b)) were determined at rest and at each workload (w during a cycle ergometer incremental exercise test. The "heart rate x blood pressure product" was calculated as (HR x SBP). The day following the incremental test, the subjects performed three repetitions of a square-wave exercise at 50% of VO(2), for the determination of pulmonary VO(2) on- and off-kinetics. VO(2) and [La](bpeak) tended to increase with training; the increases became significant at T60 or T90. HR(peak)and (HR x SBP)(peak) were unaffected by training. The time constant of the "primary" component of the VO(2) on-kinetics (tau(2)) was 46.9 +/- 17.3 s (T0), 38.1 +/- 14.2 s (T7), 34.4 +/- 12.6 s (T15), 28.8 +/- 6.8 s (T30), 30.2 +/- 8.0 s (T60), and 30.4 +/- 12.4 s (T90); a significant difference compared to T0 was observed from T15 onward. From T15 onward, tau(2) were not significantly different from values obtained (29.2 +/- 5.3 s) from a group of healthy untrained young controls ( n=7, 21.6 +/- 0.5 years). The same pattern of change as a function of training was described for the VO(2) off-kinetics. It is concluded that in 50-year-old subjects VO(2) on- and off-kinetics are more sensitive to exercise training than other physiological variables determined at peak exercise. PMID- 11889566 TI - Age-related heart rate response to exercise in heart transplant recipients. Functional significance. AB - The heart rate (HR) and O(2) uptake (VO(2)) responses to cycle ergometer exercise and the role of O(2) transport in limiting submaximal and maximal aerobic performance were assessed in 33 heart transplant recipients (HTR) [14 children (P HTR), 11 young adults (YA-HTR) and 8 middle-age adults (A-HTR)] and in 28 age matched control subjects (CTL). In 7 P-HTR ("responders") the HR response to the onset of exercise (on-response) was as fast as that of CTL, whereas in all other patients ("non-responders") the HR on-response was typical of the denervated heart. Compared with non-responder P-HTR, responder P-HTR were also characterized by a normal peak HR (177+/- 16 vs. 151+/- 25 beats/min), an equally slow time constant for the VO(2) on-response (tau: 54 +/- 11 vs. 62+/- 13 s) and a similar low (approximately 60% of that of CTL) peak VO(2) (28 +/- 7 vs. 26 +/- 10 ml/kg per min). On the other hand non-responder YA-HTR and A-HTR were characterized by a relatively low peak HR (151 +/- 21 and 144 +/- 29 beats/min, respectively), a slow tau for the on-response (63 +/- 12 and 70 +/- 11 s) and a low peak (28 +/- 7 and 19 +/- 6 ml/kg per min). In conclusion, a sizeable number of paediatric patients (responder P-HTR) may reacquire the normal HR response to exercise, both in terms of kinetics and maximal level. Despite the almost complete recovery of cardiovascular function, and, probably, oxygen delivery, both the kinetics of the VO(2) on-response and the maximal aerobic power of the responder P-HTR were similar to those of non-responder P-HTR. The latter finding is probably attributable to peripheral limitations, due to inborn and/or pharmacological muscle deterioration. PMID- 11889567 TI - Repetitive activation of postsynaptic GABA(A )receptors by rapid, focal agonist application onto intact rat striatal neurones in vitro. AB - GABA(A) receptor-mediated Cl(-) currents were evoked by rapid and short (0.75-1.5 ms), focal iontophoretic applications of GABA to proximal dendrites of cultured striatal neurones. The mean amplitude (232 +/- 21.0 pA) was in the range of large miniature inhibitory postsynaptic currents (mIPSCs), while the 10-90% rise times (3.4 +/- 0.20 ms) and the time constants of decay (60.0 +/- 8.4 ms) were three times slower. Responses to paired-pulse application of GABA at inter-pulse intervals (IPIs) of 30, 100 and 300 ms showed no depression of the second response (R2) relative to the first (R1). At short IPIs, at which R1 and R2 showed temporal summation, the net amplitude of R2 (R2(net)) was partially occluded. Lowering the iontophoretic intensity during paired-pulse application at an IPI of 30 ms reduced R1 by 69.5 +/- 3.3%, while occlusion of R2(net) decreased significantly, indicating that the receptor occupancy had been lowered. During the course of ten pulses of GABA at 10 Hz, responses declined by 8.7 +/- 3.9%, which probably reflects slow cumulative desensitization. In conclusion, rapid focal iontophoresis of exogenous GABA onto GABA(A) receptors does not result in activity-dependent depression of the postsynaptic response. This result suggests that fast desensitization is unlikely to occur during repetitive GABA(A) receptor mediated synaptic transmission in striatal neurones. PMID- 11889568 TI - Effects of temperature on adenosine A1 receptor activation in guinea pig hippocampus in vitro. AB - The effects of temperature on adenosine A1 receptor activation were studied both by electrophysiological analysis of synaptically evoked responses in CA1 neurons in guinea pig hippocampal slices, and by measuring the binding of adenosine analogues to adenosine A1 receptors in crude synaptosomes from guinea pig hippocampal neurons. Increasing the temperature of the perfusing medium from 30 degrees C to 45 degrees C attenuated the amplitude of the synaptically and the non-synaptically evoked CA1 population spikes. Bath application of 1 microM 8 cyclopentyltheophylline, an adenosine A1 receptor antagonist, did not affect non synaptically evoked CA1 population spikes, but significantly increased the amplitude of synaptically evoked population spikes in the upper range of hyperthermia (37-43 degrees C). In contrast, application of 5 microM L- N(6) phenylisopropyladenosine, an adenosine A1 receptor agonist, did not affect non synaptically evoked CA1 population spikes, but significantly decreased the amplitude of synaptically evoked population spikes in the upper range of hyperthermia. Binding assays using crude hippocampal synaptosomes showed that the affinity of adenosine A1 receptors for a radio-labeled adenosine analogue increased in response to a temperature increase. These results suggest that increased activation of adenosine A1 receptors in response to a temperature increase depresses excitatory synaptic responses in hippocampal CA1 neurons. PMID- 11889569 TI - Cumulative inactivation and the pore domain in the Kv1 channels. AB - AKv1.1a is an Aplysia Kv1 channel close to a mammalian Kv1.4. Both channels show a prominent frequency-dependent cumulative inactivation. The cumulative inactivation of AKv1.1a but not of rKv1.4 was enhanced by the patch excision. To gain structural information about the phenomenon, we examined chimeras of AKv1.1a and rKv1.4. Chimeras with the AKv1.1a pore domain displayed enhanced cumulative inactivation after patch excision. In the pore domain, eight amino acids are different between the two channels. We, therefore, constructed eight mutants of AKv1.1a (A378E, D379P, Q380T, K384Q, R406K, G409T, W411G, L414I) based on the sequence differences. All the mutants showed a similar macroscopic current decay, suggesting that N-type inactivation was not affected. The patch excision failed to enhance the cumulative inactivation in A378E, D379P, G409T, and L414I. P/C type inactivation in N-terminal deletion mutants (Delta N) became slower in A378E, D379P, G409T and L414I. Internal application of the N-terminal peptide from the ShB channel induces a frequency-dependent block of Delta N-AKv1.1a. By contrast, the block was not frequency dependent in Delta N-A378E, Delta N-D379P and Delta N-G409T. The results suggest that the positions 378, 379, 409, and perhaps 414 in the AKv1.1a backbone are involved in the stability of P/C-type inactivation and the cumulative inactivation of Kv1 channels. PMID- 11889570 TI - A purinergic signal transduction pathway in mammalian skeletal muscle cells in culture. AB - The effects of adenosine 5'-triphosphate (ATP) on human and mouse skeletal muscle fibres in primary culture were investigated. ATP-evoked changes in intracellular calcium concentration ([Ca(2+)](i)) were measured and compared with those induced by agonists of the nicotinic acetylcholine (Ach)- and P2X purinoreceptors. While ATP was effective on both myoblasts and multi-nucleated myotubes in the micromolar range, Ach failed to induce any change in [Ca(2+)](i) at early stages of development. In contrast, myofibres with peripheral nuclei showed little response to ATP but responded to Ach with a large change in [Ca(2+)](i). The responsiveness of the myotubes to Ach paralleled that to potassium. The removal of external calcium abolished the response to ATP. P2X receptor agonists mimicked the response to ATP with the order of potency being ATP>2',3'- O-(4-benzoyl) benzoyl-ATP>beta,gamma-methylene-ATP>alpha,beta-methylene-ATP. Under voltage clamp conditions ATP induced an inward current that showed little inactivation. These results are consistent with the existence of P2X receptor-mediated signal transduction pathway in cultured mammalian skeletal muscle cells. PMID- 11889571 TI - Point mutations in the pore region directly or indirectly affect glibenclamide block of the CFTR chloride channel. AB - The sulfonylurea glibenclamide is a relatively potent inhibitor of the CFTR Cl(-) channel. This inhibition is thought to be via an open channel block mechanism. However, nothing is known about the physical nature of the glibenclamide-binding site on CFTR. Here we show that mutations in the pore-forming 6th and 12th transmembrane regions of CFTR affect block by intracellular glibenclamide, confirming previous suggestions that glibenclamide enters the pore in order to block the channel. Two mutations in the 6th transmembrane region, F337A and T338A, significantly weakened glibenclamide block, consistent with a direct interaction between glibenclamide and this region of the pore. Interestingly, two mutations in the 12th transmembrane region (N1138A and T1142A) significantly strengthened block. These two mutations also abolished the dependence of block on the extracellular Cl(-) concentration, which in wild-type CFTR suggests an interaction between Cl(-) and glibenclamide within the channel pore that limits block. We suggest that mutations in the 12th transmembrane region strengthen glibenclamide block not by directly altering interactions between glibenclamide and the pore walls, but indirectly by reducing interactions between Cl(-) ions and glibenclamide within the pore. This work demonstrates that glibenclamide binds within the CFTR channel pore and begins to define its intrapore binding site. PMID- 11889572 TI - ICln channels reconstituted in heart-lipid bilayer are selective to chloride. AB - ICln is an ion channel cloned from renal epithelial cells. The reconstitution of the protein in 1,2-diphytanoyl- sn-glycero-3-phosphocholine (Diph-PC) bilayer membranes reveals potassium-selective channels, which become more chloride selective in the presence of calcium. Here we show that the ion selectivity of ICln also depends on the lipid environment in which the channels are reconstituted. Diph-PC is a synthetic lipid commonly used for reconstituting ion channels. However, since this lipid is not found in native membranes, we reconstituted the ICln ion channels in a polar heart-lipid extract. Using this lipid mixture the reconstituted ICln ion channels are chloride selective in the presence of calcium and an acidic pH. The relative ion selectivity of ICln under these conditions is similar to the cation versus anion selectivity of native ion channels activated by cell swelling. PMID- 11889573 TI - Calcium-induced calcium release participates in cell volume regulation of rabbit TALH cells. AB - Changes of cell volume and intracellular calcium concentration ([Ca(2+)](i)) in immortalized thick ascending limb of Henle's loop (TALH) cells were monitored using confocal laser scanning microscopy and fura-2 fluorescence, respectively. Reduction of the extracellular osmolarity from 600 to 300 mosmol/l induced cell swelling followed by regulatory volume decrease (RVD). Simultaneously, the [Ca(2+)](i) increased transiently. The calcium rise was not observed in calcium free solution or in the presence of nifedipine, indicating that the change was, in the first place, due to the activation of a calcium influx. Application of ATP or caffeine in isotonic solutions increased transiently the [Ca(2+)](i) which revealed the existence of stores in TALH cells sensitive to inositol-1,4,5 trisphosphate (IP(3)) and ryanodine. To examine the possibility that the calcium influx might induce calcium release, manganese quenching experiments were performed. In hypotonic calcium-free solutions, the decay of the calcium insensitive and calcium-sensitive fluorescence occurred simultaneously. In the presence of extracellular calcium however, the calcium-sensitive wavelength revealed initial calcium influx followed by a calcium release from intracellular stores. Thus, the calcium influx was a prerequisite for the calcium release. We conclude that calcium-induced calcium release participates in global calcium signalling during RVD of TALH cells. PMID- 11889574 TI - Inhibition of muscle carbonic anhydrase increases rise and relaxation times of twitches in rat skeletal muscle fibres. AB - Muscle carbonic anhydrase (CA) was inhibited in fibre bundles of the extensor digitorum longus (EDL) and soleus (SOL) muscles from rats. Isometric single twitches were recorded in the absence or presence of the CA inhibitors. The highly membrane-permeable inhibitors L-645,151, chlorzolamide (CLZ) and ethoxzolamide (ETZ) prolonged significantly the values of time-to-peak (ttp) by 5 40 ms (10-40%) in both muscles and the values of the 75% decay time (t(75%)) by 30-400 ms (13-110%) in SOL and by 9-17 ms (15-30%) in EDL and increased peak force by 20--55% in SOL and EDL. The poorly membrane-permeable inhibitors benzolamide (BZ) and acetazolamide (ACTZ) had no effects on single twitches. In CO(2)-free solution, the effects of L-645,151 on ttp, t(75%) and peak force of SOL were reduced drastically. Removal of CO(2) prolonged ttp and t(75%). In skinned fibres, ETZ and CLZ did not increase force production. Intracellular pH (pH(i)) in SOL and EDL fibres was not affected by 30-60 min exposure to CLZ, ETZ or BZ. The results of L-645,151, CLZ and ETZ on ttp, t(75%) and peak force of twitches are consistent with our hypothesis on the role of the sarcoplasmic reticulum (SR) CA. The SR-CA may mediate sufficiently fast buffering and production of H(+) in the SR that is exchanged for Ca(2+) across the SR membrane. We propose that a H(+) buffering and delivery impaired by CA inhibition slows the kinetics of Ca(2+) release and reuptake and, as a result, slows twitch ttp and t(75%). Aspects of this hypothesis await further validation. PMID- 11889575 TI - Effects of extracellular ATP on freshly isolated mouse skeletal muscle cells during pre-natal and post-natal development. AB - Extracellular adenosine 5'-triphosphate (ATP) has profound effects on membrane conductance and on the intracellular free [Ca(2+)] ([Ca(2+)](i)) in cultured skeletal muscle cells. The aim of the present study was to examine the occurrence and to characterize the properties of such responses during mammalian muscle development in vivo. The effect of ATP (0.2 mM) was tested on membrane current and [Ca(2+)](i) in freshly isolated pre- and post-natal mouse skeletal muscle cells. Pre-natal cells were from 14- to 19-day-old fetuses. In pre- and early post-natal cells, very small elevations of [Ca(2+)](i) (<50 nM) following ATP application could be detected with the fluorescent indicator fura-2. A clear subsarcolemmal rise in [Ca(2+)] was however associated to the presence of ATP, as demonstrated by increased activity of plasma membrane Ca(2+)-activated K(+) channels in cells bathed in a depolarizing, high-calcium-containing solution. In cells voltage-clamped at -80 mV in external Tyrode, ATP induced an inward current associated with an increased membrane conductance. The mean maximal amplitude of the ATP-induced current was -0.84 +/- 0.07 A/F ( n=39). The response to ATP was still present after birth, although its amplitude tended to decrease with post natal development and was completely absent in muscle cells from 3- to 6-month old mice. The ATP-induced current could be abolished reversibly by suramin. Our results suggest that, over the range of developmental stages examined, skeletal muscle cells display an ionotropic purinergic signalling pathway with functional properties qualitatively consistent with what is observed in cultured myotubes. PMID- 11889576 TI - Ionic mechanisms of regulatory volume increase (RVI) in the human hepatoma cell line HepG2. AB - We studied the effects of hypertonic stress on ion transport and cell volume regulation (regulatory volume increase; RVI) in the human tumor cell-line HepG2. Ion conductances were monitored in intracellular current-clamp measurements with rapid ion-substitutions and in whole-cell patch-clamp recordings; intracellular pH buffering capacity and activation of Na(+)/H(+) antiport were determined fluorometrically; the rates of Na(+)-K(+)-2Cl(-) symport and Na(+)/K(+)-ATPase were quantified on the basis of time-dependent and furosemide- or ouabain sensitive (86)Rb(+) uptake, respectively; changes in cell volume were recorded by means of confocal laser-scanning microscopy. It was found that hypertonic conditions led to the activation of a cation conductance that was inhibited by Gd(3+), flufenamate as well as amiloride, but not by benzamil or ethyl-isopropyl amiloride (EIPA). Most likely, this cation conductance was non-selective for Na(+) over K(+). Hypertonic stress did not change K(+) conductance, whereas possible changes in Cl(-) conductance remain ambiguous. The contribution of Na(+)/H(+)antiport to the RVI process appeared to be minor. Under hypertonic conditions an approximately 3.5-fold stimulation of Na(+)-K(+)-2Cl(-)symport was observed but this transporter did not significantly contribute to the overall RVI process. Hypertonic stress did not increase the activity of Na(+)/K(+)-ATPase, which even under isotonic conditions appeared to be working at its limit. It is concluded that the main mechanism in the RVI of HepG2 cells is the activation of a novel non-selective cation conductance. In contrast, there is little if any contribution of K(+) conductance, Na(+)/H(+) antiport, Na(+)-K(+)-2Cl(-) symport, and Na(+)/K(+)-ATPase to this process. PMID- 11889577 TI - Diet supplementation with vitamin E, vitamin C and beta-carotene cocktail enhances basal neutrophil antioxidant enzymes in athletes. AB - Exercise increases oxygen consumption and causes a disturbance of intracellular pro-oxidant-antioxidant homeostasis. Few data are available as to the cumulative effects of exercise on the antioxidant defenses of the neutrophil. We studied the effects of 90 days' supplementation with placebo or an antioxidant cocktail of vitamin E (500 mg/day) and beta-carotene (30 mg/day) and the last 15 days also with vitamin C (1 g/day) on sportsmen's basal neutrophil antioxidant defenses. We analyzed the activities of catalase, glutathione peroxidase, glutathione reductase and the activities and levels of superoxide dismutase, glutathione and glutathione disulfide in neutrophils purified from antecubital vein blood of sportsmen before and after diet supplementation. Plasma vitamin E, beta-carotene and vitamin C concentrations in the antioxidant-supplemented group were approximately 1.6, 10, and 1.2 times higher respectively than those of the placebo group. The antioxidant-supplemented group presented a significantly higher glutathione versus glutathione disulfide ratio in neutrophils (about 20%) than the placebo one. Antioxidant supplementation enhances the antioxidant enzyme activity of superoxide dismutase and catalase in neutrophils. PMID- 11889578 TI - Stimulation of TNF alpha expression by hyperosmotic stress. AB - Hyperosmotic stress is known to induce apoptotic cell death, an effect previously attributed to seemingly ligand-independent clustering of tumour necrosis factor alpha (TNF alpha) receptors. An alternative explanation for the clustering of TNF alpha receptors may be stimulation of TNF alpha production, with subsequent autocrine or paracrine stimulation of the receptors. The present study was performed to test for an effect of exposure to hyperosmotic extracellular fluid on cellular TNF alpha production. In both the macrophage cell line U937 and the B lymphocyte cell line LCL721, an increase of extracellular osmolarity to 500 mosmol/l indeed increased TNF alpha expression, an effect reversed by the p38 kinase inhibitor SB203580. In both cell types hyperosmotic stress triggered apoptosis, which in U937 cells was significantly inhibited by neutralizing antibodies against TNF alpha and by SB203580 and was similarly elicited by exogenous addition of TNF alpha. In contrast, osmotically induced apoptosis of LCL721 cells was only slightly blunted by anti-TNF alpha antibodies and rather increased by SB203580. In conclusion, through activation of p38 kinase hyperosmotic stress stimulates the expression of TNF alpha which at least in U937 macrophages may participate in the triggering of subsequent apoptotic cell death. However, the observations in LCL721 cells point to other, TNF alpha-independent, mechanisms mediating apoptotic cell death following an excessive increase of extracellular osmolarity. PMID- 11889579 TI - Potentiation of shortening and velocity of shortening during repeated isotonic tetanic contractions in mammalian skeletal muscle. AB - The purpose of this study was to investigate the enhancement of shortening and of the velocity of shortening during repeated incompletely fused isotonic tetanic contractions. The medial gastrocnemius muscle of anesthetized rats was isolated in situ and the motor nerve stimulated with supramaximal pulses, 50 micros duration, at optimal length. Estimated maximal velocity of shortening (V(max)) was 210 +/- 6 mm x s(-1) (mean +/- SEM). Repeated incompletely fused tetanic contractions (three pulses at 80 Hz) resulted in initial shortening which was 1.5 +/- 0.1 mm, and this increased to 2.7 +/- 0.2 mm after 7 s of 4 s(-1) contractions. Peak velocity of shortening for intermittent 80 Hz stimulation increased from 60.5 +/- 4 mm x s(-1) to 91.8 +/- plus minus 6 mm x s(-1). The initial velocity of shortening for 80 Hz stimulation was substantially less than the velocity of shortening observed with 200 Hz stimulation, but increased to 72 +/- 3% of the load-specific value for 200 Hz stimulation. Myosin regulatory light chain phosphorylation increased from 11.1 +/- 1.5% at rest to 32.9 +/- 5.4% after 4 s of intermittent 80 Hz stimulation and 50.4 +/- 8.8% after 7 s ( P<0.01). The ascending limb of the length-force relationship was determined with tetanic contractions, 200 Hz for 100 ms. At the length corresponding to peak shortening after 7 s of repeated 80 Hz contractions, the maximal isometric force was five times greater than the isotonic load. The rate of phosphorylation was sustained from 4 to 7 s, but the rate of increase in shortening and velocity decreased. The slower rate of change in shortening and velocity may be due to approaching maximal velocity for this short duration of contraction, but is not due to slowing of the rate of phosphorylation of the myosin regulatory light chains. PMID- 11889580 TI - Activity modes and modulation of the peptide-gated Na(+) channel of Helix neurones. AB - Unitary currents activated by steady-state applications of FMRFamide analogues have been recorded in outside-out patches from the C2 and F2 neurones of Helix aspersa. The bimodal conductance distribution ranged from 4 pS to 6 pS in patches in each neurone. Application of up to 10 microM cytochalasin B or D to 12 of the 41 inside-out patches tested stimulated activity of the larger conductance events and/or increased the amplitude of smaller conductance events, indicating a cytoskeletal influence on the channels. FMRFamide-gated currents were blocked by external Ca(2+) and Mg(2+), both with an IC(50) of about 1 mM. During continuous application of FMRFamide, clustering of events and/or the appearance of a mode of activity with a lower probability of being open (P(open)) suggested two different types of desensitization. More rarely, exceptionally high P(open) activity with low FMRFamide concentrations was seen. In some cases, there were long periods of high activity during which the channels gated normally with regular short sojourns in the fully closed state. In others, isolated long openings were seen which interrupted the normal gating pattern of the channels. Amiloride and its analogue 5-(N-ethyl- N-isopropyl) amiloride (EIPA) consistently caused channel block, but in some patches with a low background activity an increased P(open) was also observed in the presence of each drug. PMID- 11889581 TI - The role of KCNQ1/KCNE1 K(+) channels in intestine and pancreas: lessons from the KCNE1 knockout mouse. AB - KCNE1 (IsK, minK) co-assembles with KCNQ1 (KvLQT1) to form voltage-dependent K(+) channels. Both KCNQ1 and KCNE1 are expressed in epithelial cells of gut and exocrine pancreas. We examined the role of KCNQ1/KCNE1 in Cl(-) secretion in small and large intestine and exocrine pancreas using the KCNE1 knockout mouse. Immunofluorescence revealed a similar basolateral localization of KCNQ1 in jejunum and colon of KCNE1 wild-type and knockout mice. Electrogenic Cl(-) secretion in the colon was not affected by gene disruption of KCNE1; in jejunum forskolin-induced short-circuit current was some 40% smaller but without being significantly different. Inhibition of KCNQ1 channels by 293B (IC(50) 1 micromol l(-1)) and by IKS224 (IC(50) 14 nmol l(-1)) strongly diminished intestinal Cl(-) secretion. In exocrine pancreas of wild-type mice, KCNQ1 was predominantly located at the basolateral membrane. In KCNE1 knockout mice, however, the basolateral staining was less pronounced and the distribution of secretory granules was irregular. A slowly activating and 293B-sensitive K(+) current was activated via cholinergic stimulation in pancreatic acinar cells of wild-type mice. In KCNE1 knockout mice this K(+) current was strongly reduced. In conclusion intestinal Cl(-) secretion is independent from KCNE1 but requires KCNQ1. In mouse pancreatic acini KCNQ1 probably co-assembled with KCNE1 leads to a voltage-dependent K(+) current that might be of importance for electrolyte and enzyme secretion. PMID- 11889582 TI - Bilateral coordination of inspiratory neurones in the rat. AB - Inspiratory activity on the left and right sides must be coordinated to be effective. We used cross-correlation to examine the hypothesis that the coordination of left and right medullary inspiratory neurones is produced by excitation from common sources and by midline-crossing excitatory connections among these neurones. In adult rats, a total of 185 contralateral pairs of inspiratory neurones ( n=370) were recorded extracellularly, and classified, according to their firing pattern, as augmenting ( n=262), constant ( n=82) or decrementing ( n=26). Of the 262 augmenting inspiratory neurones, 98 were classified as phrenic premotor neurones by cross-correlation with phrenic nerve discharge. The 185 cross-correlograms showed little evidence of common activation, or midline-crossing excitatory connections. Of the 45 cross correlograms for pairs of augmenting neurones, only 4 (approximately equal to 9%) indicated a common activation, and only one a monosynaptic connection. Of the 45 for pairs of augmenting and phrenic premotor neurones, only 9 (20%) showed a common activation, and only 2 a monosynaptic excitatory connection. Of the 19 pairs of phrenic premotor neurones, 5 from the same rat showed high-frequency oscillations, and 1 a monosynaptic excitatory connection. Cross-correlograms for pair combinations of other types of neurones also exhibited few features. We suggest that, in the adult rat, although both common activation and excitatory cross-connections exist as a means for coordinating left and right ventral group inspiratory neurones to the same respiratory rhythm, they are insufficient to account for it. PMID- 11889583 TI - Intracellular ATP measured with luciferin/luciferase in isolated single mouse skeletal muscle fibres. AB - The firefly luciferin/luciferase reaction was utilized to monitor intracellular ATP concentration ([ATP](i)). Single fibres of mouse skeletal muscle were dissected and injected with luciferase. Luciferin was added to the perfusate and light emission from the fibres was monitored as an indication of [ATP](i). Inhibition of oxidative phosphorylation with cyanide and anaerobic glycolysis with iodoacetate caused light emission to fall to zero within 10 min and the fibres developed a rigor contraction. Inhibition of creatine kinase with 2,4 dinitro-1-fluorobenzene produced a small transient fall in light emission in association with each tetanus. Muscle fibres were fatigued by repeated tetani and 5/12 fibres showed a fall in light emission in the late phase of fatigue. If fibres were allowed to recover from fatigue in the absence of glucose and then restimulated in the absence of glucose they fatigued much more rapidly. However, such fibres showed no obvious change in light emission. We conclude that the luciferin/luciferase system can be used to monitor [ATP](i) in functioning single skeletal muscle cells. A depletion of global [ATP](i) is not observed in all fatiguing fibres and cannot be the sole cause of the final phase of fatigue. PMID- 11889584 TI - pH gating of lens fibre connexins. AB - Chemical gating of gap junction channels by intracellular pH may be an important mechanism for the physiological regulation of cell-cell coupling. In the ocular lens, pH gating of gap junction channels has been implicated as a possible cause of cataract in diabetics. To address this question further, I determined the pH dependence of the rat connexin (Cx)-46 and ovine Cx49 in transfected HeLa cells using the pH-clamp technique during dual whole-cell recording. pH gating for both connexins was fast and reversible. The apparent p K(a) (p K(a,app)) was 6.66 +/- 0.01 and the Hill coefficient ( n) 6.8 +/- 1.8 for Cx49, and for Cx46 6.8 +/- 0.01 and 2.2 +/- 0.15, respectively. C-terminal truncation of Cx46 by 163 aa did not abolish the pH sensitivity but shifted the p K(a,app) to 6.6 +/- 0.01. This finding is inconsistent with the ball-and-chain model proposed for Cx43. Voltage gating of Cx46 channels was also not altered by truncation or acidic pH, indicating that the two gating mechanisms are functionally and possibly structurally separate. The data also imply a significant role of pH gating for lens pathophysiology. For the normal pH range in the lens cortex (pH 6.8-7.2) most gap junction channels will be open. However, mild acidification will reduce gap junctional coupling significantly, especially for Cx50 channels. Localized closure of gap junction channels will disrupt lens transport and thus may contribute to the tissue damage observed in diabetic lenses. PMID- 11889585 TI - Submaximal exercise in healthy volunteers: the relationship between gastric mucosal and systemic energy status. AB - The effect of exercise on gastric mucosal energy status has not been fully elucidated. The aim of this study was to evaluate the impact of submaximal cycling on gastric mucosal energy balance and its relationship to changes in systemic energy status. Ten healthy volunteers (age 20-40 years) were investigated at rest (BL), during 30 min of submaximal exercise (E) on bicycle ergometry and during the 30 min after the completion of cycling. Gastric mucosal PCO(2) ( P(gm)CO(2)) was measured by air tonometry at 10-min intervals and the gastric mucosal-arterial PCO(2) difference ( PCO(2)gap) was calculated. Hemodynamics, arterial blood gases, lactate and pyruvate were also measured. PCO(2)gap significantly increased throughout exercise [BL: 0.2 kPa (median), -0.1 0.6 kPa (25th-75th percentiles); E(10 min): 1.0 kPa, 0.8-1.7 kPa; E(20 min): 1.35 kPa, 0.8-1.8 kPa; E(30 min): 1.5 kPa, 0.9-2.0 kPa]. The early changes in PCO(2)gap ( PCO(2)gap at E(10 min) minus PCO(2)gap at BL) correlated significantly and positively with corresponding changes in arterial lactate ( r(2)=0.58, P<0.05) and lactate-to-pyruvate ratio ( r(2)=0.72, P<0.05). On recovery, all metabolic parameters normalized within 30 min. We conclude that submaximal cycling in volunteers leads to the early derangement of gastric mucosal energy balance. The time course of PCO(2)gap parallels changes in systemic energy status. PMID- 11889586 TI - Long-lasting adverse effects of prenatal hypoxia on developing autonomic nervous system and cardiovascular parameters in rats. AB - To determine whether prenatal hypoxia increases the risk of developing cardiovascular disorders as an adult and, if so, the identity of the cell mechanisms involved in such dysfunction, we evaluated the sympathoadrenal system and central areas related to cardiovascular events during development and the cardiovascular parameters in adults. Pregnant rats were exposed to hypoxia (10% oxygen) from embryonic day (E) 5 to E20 and the offspring studied at 1, 3, 9 and 12 weeks of age for neurochemistry and at 12 weeks of age for cardiovascular analysis. In the 1-, 3- and 9-week-old offspring, the levels and utilization of catecholamines were reduced in sympathetic ganglia, in target organs, in adrenals and in the rostral part of the A2 cell group in the nucleus tractus solitarius, but were increased in the locus coeruleus. In the 12-week-old adult offspring, the lowered autonomic nervous activity was restricted to cardiac-related structures, i.e. the stellate ganglion, heart and adrenals. In adult rats, prenatal hypoxia did not affect the cardiac parameters under resting conditions but increased blood pressure and the variability of blood pressure and heart rate under stress conditions. The altered metabolic activity of the sympathoadrenal system and related central areas during development and at adulthood for most structures might be part of the potential mechanisms contributing to cardiovascular disorders in adults. PMID- 11889587 TI - Effects of transcutaneous short-term electrical stimulation on M. vastus lateralis characteristics of healthy young men. AB - Fifteen healthy, untrained male subjects (mean age +/- SD, 22 +/- 5 years) were used to examine the plasticity of myosin heavy chain phenotype, size, oxidative capacity and capillarization of skeletal muscle fibre types with short-term electrical stimulation (ES). Ten subjects were electro-stimulated on both quadriceps muscles with a frequency of 45-60 Hz, with 12 s of stimulation followed by 8 s of recovery for a total of 30 min per day, 3 days per week for 6 weeks. The remaining five subjects served as controls. Two vastus lateralis muscle biopsy samples were removed from each subject before (week 0) and after (week 6) ES training. A standardized exercise test on a cycle ergometer was performed by each subject before and after the experimental period and several indicators of whole-body aerobic capacity were estimated. The so-called electromyographic threshold was also determined during the tests. Muscle biopsy samples were analysed by electrophoresis, immunohistochemistry and quantitative histochemistry. Myosin heavy chain (MHC) composition, muscle fibre type distribution, fibre areas, oxidative capacity and capillaries of each fibre type were estimated. Muscular changes with ES revealed an increase of fibres expressing MHC-IIA, and a decrease of fibres expressing MHC-IIX and MHC-I, as well as an increase of the oxidative capacity and mean number of capillaries of fast-twitch (type II) fibres with minimal muscle fibre hypertrophy. These adaptations seem related to a bi-directional transformation from both MHC isoforms I and IIX towards the MHC-IIA isoform. The aerobic performance and electromyographic variables at the whole-body level were not altered by ES. These results indicate that the particular short-term ES training protocol tested in the present study induces significant adaptations in histochemical and metabolic machineries of human skeletal muscle. The results also offer new perspectives for realistic applications of ES in various clinical situations and sport training. PMID- 11889588 TI - The role of bicarbonate in regulatory volume decrease (RVD) in the epithelial derived human breast cancer cell line ZR-75-1. AB - This study investigates the mechanisms involved in the regulatory volume decrease (RVD) in ZR-75-1 epithelial-derived human breast cancer cells. Cell volume changes were measured during osmotic shock using video imaging. In HEPES-buffered hypotonic solutions no RVD was observed; however, RVD was observed in HCO(3)(-) buffered hypotonic solutions. Inhibition of RVD by 10 microM tamoxifen and 100 microM DIDS (inhibitors of volume-regulated anion channels; VRAC) and 2 mM TEA(+) (inhibitor of K(+) channels) indicates a role for these channels. In HCO(3)(-) buffered Cl(-)-free solutions RVD was partially abolished indicating that HCO(3)( ) efflux can support RVD but also may have another role. Further experiments investigated whether HCO(3)(-) assists in the accumulation of Cl(-) via Cl(-) HCO(3)(-) exchange. Regulatory volume increase (RVI) was also HCO(3)(-)-dependent and was inhibited by 500 microM DIDS and 10 microM 5-( N, N-dimethyl)-amiloride (DMA) indicating a role for coupled Cl(-)-HCO(3)(-) and Na(+)-H(+) exchange. Finally, in the presence of 10 microM DMA, RVD was partially inhibited providing further evidence for a role of Cl(-)-HCO(3)(-) exchange. Thus RVD in ZR-75-1 cells involves the activation of VRAC and K(+) channels. RVD is HCO(3)(-) dependent and HCO(3)(-) efflux through VRAC appears to contribute directly to RVD. HCO(3)(-), however, also has another role in facilitating Cl(-) accumulation via Cl(-)-HCO(3)(-) exchange. PMID- 11889589 TI - Influence of voltage and extracellular Na(+) on amiloride block and transport kinetics of rat epithelial Na(+) channel expressed in Xenopus oocytes. AB - We expressed the three subunits of the epithelial amiloride-sensitive Na(+) channel (ENaC) from rat distal colon heterologously in oocytes of Xenopus laevis and analysed blocker-induced fluctuations in current using conventional dual microelectrode voltage-clamp. To minimize Na(+) accumulation we performed all experiments in low-Na(+) solutions (15 mM). Noise analysis revealed that control or ENaC-injected oocytes did not exhibit spontaneous relaxation noise. However, in ENaC-expressing oocytes, amiloride induced a distinct Lorentzian component in the power density spectra. With three amiloride concentrations and a linear analysis of the respective changes in the corner frequency f(c) (2 pi f(c) plot) we determined the rate constants k(on) and k(off) for the amiloride-ENaC interaction. At a clamp potential (V(m)) of -60 mV k(on) was 80.8 +/- 5.1 microM( 1) s(-1) and k(off) 15.4 +/- 4.2 s(-1). The half-maximal blocker concentration (K(mic,ami)) was 0.19 microM (V(m)=-60 mV). While k(on) was voltage-independent in the range -50 to -100 mV, k(off) and K(mic,ami) decreased significantly with increasing membrane hyperpolarization, resulting in an increased affinity of amiloride for its binding site on ENaC. Increasing extracellular [Na(+)] ([Na(+)](o)) led to saturation of ENaC. Subsequent noise analysis revealed that single-channel current increased non-linearly with [Na(+)](o) and that saturation was not due to a reduction in the number of open channels. The apparent affinity of Na(+) for its binding site on the channel was voltage dependent and increased with hyperpolarization. Noise analysis revealed that k(on) and k(off) for amiloride decreased with increasing [Na(+)](o), while the affinity of the amiloride-binding site did not change. These findings show that the affinity of rat intestinal ENaC for amiloride is voltage dependent and is influenced non competitively by [Na(+)](o), indicating that Na(+) and amiloride do not compete for the same binding site at the channel. PMID- 11889590 TI - Monovalent cation permeability and Ca(2+) block of the store-operated Ca(2+) current I(CRAC )in rat basophilic leukemia cells. AB - Like voltage-operated Ca(2+) channels, store-operated CRAC channels become permeable to monovalent cations in the absence of external divalent cations. Using the whole-cell patch-clamp technique, we have characterized the permeation and selectivity properties of store-operated channels in the rat basophilic leukemia (RBL-1) cell line. Store depletion by dialysis with InsP(3) and 10 mM EGTA resulted in the rapid development of large inward currents in Na(+)- and Li(+)-based divalent-free solutions. Cs(+) permeated the channels poorly (P(Cs)/ P(Na)=0.01). Trimethylamine (TMA(+)), tetramethylammonium (TeMA(+)), tetraethylammonium (TEA(+)), N-methyl- D-glucamine (NMDG(+)) and TRIS(+) were not measurably permeant. NH(4)(+) was conducted well. We estimated the minimum pore diameter under divalent-free conditions to be between 0.32 nm and 0.55 nm. When cells were dialysed with buffered Ca(2+) solution and I(CRAC) activated by application of thapsigargin, P(Cs)/ P(Na) was still low (0.08). Outward currents through CRAC channels were carried by intracellular Na(+), K(+) and, to a much lesser extent, by Cs(+). Currents were unaffected by dialysis with Mg(2+)-free solution. The Na(+) current was inhibited by external Ca(2+) (half-maximal blocking concentration of 10 microM). This Ca(2+)-dependent block could be alleviated by hyperpolarization. The monovalent Na(+) current was voltage dependent, increasing as the holding potential depolarized above 0 mV. Our results suggest that CRAC channels in RBL-1 cells have a smaller pore diameter than voltage-operated Ca(2+) channels, discriminate between Group I cations, and differ markedly in their selectivity from CRAC channels reported in lymphocytes. PMID- 11889591 TI - Spontaneous nerve activity and sensitivity in catfish ampullary electroreceptor organs after tetanus toxin application. AB - The functioning of electroreceptor organs of Ictalurus sp. was investigated by inhibiting synaptic transmission by the administration of tetanus toxin in vitro. A piece of Ictalurus skin of about 20 mm diameter was mounted in an Ussing-type chamber. After establishing the normal functioning of the organ, tetanus toxin (TeTx) was applied basolaterally for 150 min in 66.7 pM and 400 pM concentrations, while the single unit nerve activity was recorded extracellularly. Spontaneous spike activity and the sensitivity of the electroreceptor organs were measured. The results show that TeTx reduces sensitivity to less then 20% of its original value, whereas the spontaneous activity is unaffected by the treatment. This indicates that the afferent nerve is capable of generating impulses independent of receptor cell neurotransmitter release. In the discussion we suggest two alternative mechanisms for the emergence of the spontaneous spike activity. PMID- 11889592 TI - Post-ischaemic changes in the response time of oxygen consumption to demand in the isolated rat heart are mediated partly by calcium and glycolysis. AB - This study examined whether different durations of ischaemia (I) and reperfusion (R) altered the kinetics of O(2) consumption-to-demand matching and the contribution of changes in calcium and metabolic pathways to possible alterations. The response time of mitochondrial O(2) consumption (t(mito)) to a step in heart rate in isolated rat hearts was used as index for the response time of O(2) consumption-to-demand matching. At baseline, t(mito) was 8.9 +/- 0.4 s for all groups. At 5 min reperfusion, after both reversible (I=5 or I=15 min) or irreversible (I=25 min) ischaemia, matching was accelerated (t(mito) relative to baseline: 53 +/- 8%, 64 +/- 8%, 51+/- 6% and 100 +/- 5% for I=5, 15, 25 min and control). At late reperfusion (>30 min), reversible ischaemia resulted in a slowing of the matching, whereas after irreversible ischaemia t(mito) recovered to control values (156 +/- 16%, 153 +/- 13%, 92 +/- 7%, 114 +/- 6%, for I=5,15, 25 min and control, respectively). High perfusate Ca(2+) mimicked (t(mito): 44 +/ 11%), whereas blocking mitochondrial Ca(2+) uptake attenuated the acceleration observed at early reperfusion (t(mito): 7 +/- 5%). Replacing glucose with substrates used downstream of glycolysis (11 mM lactate or 11 mM pyruvate) abolished the reversible ischaemia-induced slowing of the matching at late reperfusion. It is concluded that I/R-induced changes in the kinetics of O(2) consumption-to demand matching depend critically on the duration of ischaemia and reperfusion. The data indicate that I/R-induced increases in Ca(2+) may, at least partly, explain the faster kinetics at early reperfusion, whereas I/R-induced increases in glycolysis from exogenous glucose result in slower matching of O(2) consumption-to-demand at late reperfusion. PMID- 11889593 TI - Neuroendocrine carcinomas of the prostate and urinary bladder: a diagnostic and therapeutic challenge. AB - This review addresses the various morphological, immunohistochemical and cell kinetic aspects of pure and mixed neuroendocrine carcinomas of the prostate and urinary bladder and of carcinomas with focal neuroendocrine differentiation. It is important that neuroendocrine tumours of the prostate and urinary bladder be clearly distinguished from their nonneuroendocrine counterparts because of differences in treatment and prognosis. In the case of high-grade neuroendocrine carcinomas, early diagnosis and initiation of appropriate chemotherapy may increase survival and potentially induce complete remission in individual cases. PMID- 11889595 TI - Retroviral transduction of splice variant Brca1-Delta11 or mutant Brca1-W1777Stop causes mouse epithelial mammary atypical duct hyperplasia. AB - We have investigated the effects of the expression of wild-type and mutant Brca1 alleles on the murine mammary gland morphogenesis and carcinogenesis. Primary cultures of mammary cells from BALB/cByJIco mice were infected with recombinant Babe Puro retroviruses expressing lacZ, full-length Brca1, splice variant Brca1 Delta11, or mutant Brca1-W1777Stop alleles. Infected cells were reinjected into the mammary fat pad of a syngeneic virgin mouse whose endogenous epithelium had previously been removed. Four months after reinjection, nulliparous and postlactating mice were checked for the reconstitution of the mammary gland. Stable expression of beta-galactosidase was observed in the ducts formed by epithelial mammary cells infected with Babe Puro/ lacZ retrovirus. Epithelial mammary cells transduced with full-length Brca1 developed normally, whereas those transduced with Brca1-Delta11 or Brca1-W1777Stop formed atypical duct hyperplasia associated with reduced branching. These results suggest that ectopically expressed splice variant Brca1-Delta11 and mutant Brca1-W1777Stop have dominant negative effects. PMID- 11889596 TI - Microcalcifications of breast cancer and atypical cystic lobules associated with infiltration of foam cells expressing osteopontin. AB - We studied the pattern of calcification and the expression of osteopontin protein and mRNA by the histiocytes and noninvasive carcinoma cells of 20 breast cancers and the histiocytes and atypical ductal cells of sixteen cases of atypical cystic lobules. Ten breast cancers showed low-grade cribriform carcinoma in situ containing secretory material; the remaining ten cancers displayed high-grade carcinoma in situ with central necrosis characteristic of comedo type carcinoma. Hematoxylin-eosin and Kossa staining revealed calcium hydroxyapatite calcifications in 80% of cribriform type carcinomas, 50% of comedo type carcinomas, and 56% of atypical cystic lobules. Immunohistochemical staining demonstrated osteopontin protein in intraluminal secretory material, necrotic debris, or stroma, and in histiocytes in all the Kossa-positive carcinomas and atypical cystic lobules. In situ hybridization revealed osteopontin mRNA mainly in the histiocytes and especially in those near the calcifications. In two cases, rare carcinoma cells contained osteopontin protein and mRNA. The close relation between hydroxyapatite crystals and osteopontin-producing histiocytes suggests that osteopontin plays a role in the biomineralization that occurs in certain noninvasive breast cancers and atypical cystic lobules. The differences in the morphology of the calcifications and the intraductal contents suggest that the mechanism leading to the osteopontin production might vary depending on the underlying lesion. PMID- 11889594 TI - Spindle cell lipoma-like tumor, solitary fibrous tumor and myofibroblastoma of the breast: a clinico-pathological analysis of 13 cases in favor of a unifying histogenetic concept. AB - We reviewed the clinico-pathological features of a series of 13 cases of benign spindle stromal tumors (BSSTs) of the breast relating to a basic common theme consisting of a well-circumscribed proliferation of vimentin+/CD34+/BCL-2+/CD99+ spindly to oval-epithelioid cells, variably arranged in haphazard to short fascicular growth pattern, with interspersed thick or thin collagen bands. Morphological variations included atypical mono- or multi-nucleated cells in five cases and a mature lipomatous tumor component, varying from focal to prominent, in eight cases. Based on morphological and immunophenotypical features, a distinction was made between two main subtypes of these tumors--fibroblastic and myofibroblastic. The former subtype included two cases respectively represented by a typical solitary fibrous tumor (SFT) and a neoplasm labeled "spindle-cell lipoma (SCL)-like tumor", closely reminiscent of soft tissue SCL. Both tumors had cells with fibroblastic-like appearance, haphazardly arranged and immunoreactive for vimentin, CD34, BCL-2, and CD99. The latter subtype, comprised nine cases exhibiting evidence of myofibroblastic differentiation (desmin and alpha-smooth muscle actin) which were classified as myofibroblastomas (MFBs). The remaining two cases were defined as "mixed BSSTs", having typical features of diverse neoplasms, respectively represented by a case of MFB with focal SFT and pleomorphic/SCL-like areas, and SFT with focal MFB-like component. The common basic morpho-immunophenotypical features, the possibility that both fibroblastic and myofibroblastic tumors may contain an additional mature lipomatous component, and the existence of hybrid stages (mixed BSSTs) strongly support the view that such tumors belong to the same category of lesions. We postulate that the precursor of all these neoplasms is the vimentin+/CD34+ cells of the mammary stroma, the well-known inherent plasticity of which to differentiate toward several mesenchymal lines, provides the explanation for the phenotypic heterogeneity of these neoplasms. Accordingly, the encompassing term "benign spindle stromal tumors of the breast" is advocated for such tumors. PMID- 11889597 TI - Loss of nuclear BRCA1 localization in breast carcinoma is age dependent. AB - Abstract. Patients carrying a germ line mutation in the BRCA1 gene are predisposed to breast cancer. Somatic BRCA1 mutations were almost never reported in sporadic breast tumors, but several authors have described a decrease in BRCA1 mRNA and protein. In order to further investigate the possible role of BRCA1 in sporadic breast cancer, an improved immunohistochemical protocol was developed and applied on tissue sections obtained from 102 cancer patients belonging to two nonoverlapping age groups (less than 40 and more than 60 years). Based on the obtained BRCA1 specific nuclear staining we could distinguish two categories of breast cancer. The staining was present in almost all the nuclei of the normal and the cancer cells in about 50% of young (less than 40 years old) as well as older patients (more than 60 years old). Thus, BRCA1 does not seem to be involved in the genesis of these cancers. In the second category of patients, either a fraction of, or all tumor cells showed no nuclear staining. In this category, no nuclear BRCA1 staining could be observed in the tumor cells of 14 (27%) young and 3 (6%) older patients. Among six young patients bearing a breast tumor showing no BRCA1 nuclear staining at all, one was found to carry a BRCA1 germline mutation. Taken together, our results suggest that the molecular pathway in which the BRCA1 protein participates is disturbed in about 50% of sporadic breast cancer, this effect being more pronounced in tumors of young patients. PMID- 11889598 TI - Prediction of recurrence and nucleolar features in node-negative breast carcinoma, ductal type, grade II. An ultrastructural study. AB - One-third of patients with node-negative breast carcinoma develop a local recurrence that will decisively influence their outcome. We conducted this quantitative ultrastructural study to determine the value of nucleolar features in predicting recurrence of node-negative breast carcinomas. Forty-three node negative infiltrating duct carcinomas were examined by means of electron microscopy. The number of cells with three or more nucleoli (N3), percentage of nucleoli contacting the nuclear membrane (NC), patient age, tumor size, grading, treatment, follow-up time, development of recurrence, and status were recorded. There were no differences in patient age, tumor grade, or modality of surgical treatment. Significant differences were found among patients with and without recurrence for N3 ( P=0.02), NC ( P=0.01), and tumor size ( P=0.01). For N3, the positive predictive value (PPV) was 64.3, and the negative predictive value (NPV) was 72.4 (sensitivity=52.9%; specificity=80.8%; P=0.04). For NC, the PPV was 62.5, and the NPV was 74.1 (sensitivity=58.8%; specificity=76.9%; P=0.02). For tumor size, the PPV was 66.7, and the NPV was 64.9 (sensitivity=23.5%; specificity=92.3%; P not significant). Using logistic regression analysis, independent predictive value was shown for N3, NC, and tumor size. The number of nucleoli per cell and the number of cells with nucleoli in contact with the nuclear membrane have independent predictive value for the development of recurrence in lymph node-negative infiltrating duct carcinoma of the breast. Although electron microscopy is a highly accurate method, improving the precision of light microscopic techniques would allow more universal assessment of these nucleolar features. PMID- 11889599 TI - Relationship of p21 (waf1/cip1) and differentiation in chondrosarcoma cells. AB - The histological grade of chondrosarcoma correlates well with their clinical behavior and with the patient's survival duration. We have previously demonstrated that p21 was expressed in the hypertrophic chondrocytes of the growth plate. To assess the relationship of p21 (waf1/cip1) to cell differentiation in chondrosarcoma, we examined the p21 expression in 14 cases of chondrosarcoma immunohistochemically and the induction of p21 by insulin-like growth factor-I (IGF-I) during cell differentiation in SW1353 chondrosarcoma cells. p21 immunoreactivity was seen in well-differentiated chondrosarcoma cells and was mutually exclusive with MIB1 reactivity in grade-1 chondrosarcoma. In vitro, the proteoglycan synthesis of SW1353 cells was increased by IGF-I in a dose-dependent manner. However, cell proliferation was not markedly stimulated. Overexpressions of p21 mRNA and p21 protein in SW1353 cells were induced by IGF-I 100 ng/ml. Our results suggested that the p21 expression was directly related to tumor differentiation and that the p21 expression was an important mediator for IGF-I in chondrosarcoma cells. PMID- 11889600 TI - Nerve fibers in tumors of the human urinary bladder. AB - Abstract. Exophytic tumors of the urinary bladder were examined by means of transmission electron microscopy for the presence of neural tissue because, as yet, there has been hardly any discussion of a neuronal component in the biology of neoplasms. In the stroma and rarely in the epithelium of bladder tumors, fine nerve strands were found to be irregularly distributed. These strands comprised one to a maximum of five axons containing predominantly colocalized clear and dense-core vesicles. Immunohistochemistry revealed some nerve-like structures showing vasoactive intestinal neuropeptide (VIP) reactivity. This response and the combination of vesicle types indicate that parasympathetic cholinergic neurons contribute to the innervation of the tumors. Thus, a morphological basis for neuronal influence on the behavior of such tumors has been demonstrated. PMID- 11889601 TI - CD34+ fibrocytes in invasive ductal carcinoma, ductal carcinoma in situ, and benign breast lesions. AB - The present study was undertaken in order to elucidate the question of whether the distribution of stromal CD34+ fibrocytes and smooth muscle actin (SMA) reactive myofibroblasts differs between benign and malignant lesions of the breast. We investigated a total of 31 ductal carcinomas and 27 specimens with benign lesions of the breast (ductal hyperplasia, sclerosing adenosis, fibroadenoma, phyllodes tumor) and compared the distribution of CD34+ fibrocytes and SMA-reactive myofibroblasts. The stroma of normal breast tissue contained CD34+ fibrocytes, whereas SMA-reactive myofibroblasts were absent. All benign breast lesions exhibited stromal CD34+ fibrocytes and few lesions (fibroadenomas and phyllodes tumor) showed additional SMA-reactive myofibroblasts. In invasive breast cancer the stroma was devoid of CD34+ fibrocytes but a varying number of stromal SMA-reactive myofibroblasts was detectable. In the setting of the present study the loss of CD34+ fibrocytes was specific for invasive breast cancer and ductal carcinoma in situ, whereas SMA-reactive myofibroblasts were observed in different benign and malignant lesions. These findings may be helpful tools in distinguishing benign breast lesions (e.g., sclerosing adenosis) from invasive breast cancer and in characterizing stromal remodeling associated with invasive cancer. PMID- 11889602 TI - Mucins MUC1, MUC2, MUC5AC and MUC6 expression in the evaluation of differentiation and clinico-biological behaviour of gastric carcinoma. AB - Normal gastric mucosa expresses MUC1 and MUC5AC in foveolar epithelium and MUC6 in mucous neck cells of the body and deep glands of the antrum. Several studies have reported aberrant expression of an under-glycosylated form of MUC1, decreased expression of "gastric" mucins and aberrant expression of "non-gastric" mucins in gastric carcinoma. In this study, we analysed the expression profile of mucins MUC1, MUC2, MUC5AC and MUC6 in 94 gastric carcinomas using immunohistochemistry. Our results showed that: (1) mucin expression is associated with tumour type (MUC5AC with diffuse and infiltrative carcinomas and MUC2 with mucinous carcinomas) but not with the clinico-biological behaviour of the tumours; and (2) mucin expression is associated with tumour location (MUC5AC with antrum carcinomas and MUC2 with cardia carcinomas), indirectly reflecting differences in tumour differentiation according to tumour location. PMID- 11889604 TI - SS1 Helicobacter pylori disrupts the paracellular barrier of the gastric mucosa and leads to neutrophilic gastritis in mice. AB - Helicobacter pylori induces severe neutrophilic infiltration in the lamina propria of the stomach, which leads to gastritis in humans. The possible involvement of a paracellular route for bacterial nutrients and etiologic agents that may play an important role in colonization of the bacteria and cause gastritis has been suggested. To study the functions of the paracellular barrier of gastric surface epithelium, SS1, a strain of H. pylori adapted to the murine stomach, was inoculated into the stomachs of C57BL/6 mice. At 4 months after inoculation, SS1 had achieved a high level of colonization (10(6)-10(7) colony forming units/g tissue) associated with neutrophilic infiltration in the lamina propria of the junctional zone. Disruption of the paracellular barrier was observed in the SS1-infected stomachs, as revealed by the invasion of a lanthanum tracer into the paracellular space of the surface epithelium. Only 2% of junctions were permeable in control stomachs, whereas 72% of the paracellular barrier was disrupted in the SS1-infected gastric epithelia. Furthermore, distribution of tight junction-related molecules such as 7H6 antigen, occludin, and cortical actin was affected in the surface epithelium by SS1 infection. The linear expression pattern of occludin was disrupted and became irregular or punctuated. The 7H6 antigen accumulated as aggregates in the apical portion of the surface epithelium and cortical actin became irregular and punctuated. Taken together, these results indicate that infection by SS1 directly or indirectly caused an increase in paracellular permeability and altered the localization of tight junction-related molecules of the gastric surface epithelium. This observation suggests that the paracellular pathway may play a significant role in establishing H. pylori-induced gastritis in the clinical setting. PMID- 11889603 TI - Mucins as key molecules for the classification of intestinal metaplasia of the stomach. AB - Mucins and mucin-associated carbohydrates have a distinct expression pattern that can be modified under pathological conditions. Normal gastric mucosa expresses MUC1 and MUC5AC in foveolar epithelium and MUC6 in the glands. Lewis type-1 chain antigens (Le(a) and Le(b)) are expressed in foveolar epithelium, whereas Lewis type-2 chain antigens (Le(x) and Le(y)) are expressed in the glands. In this study we used monoclonal antibodies to evaluate the pattern of mucins and Lewis type-1 carbohydrates in intestinal metaplasia (IM) and compared it with IM types determined using histochemistry. In type-I or complete IM we found expression of MUC2 intestinal mucin and decreased/absent expression of MUC1, MUC5AC and MUC6. In type-II/III or incomplete IM there was co-expression of MUC2 and the mucins expressed in the stomach. No major differences were detected among the three IM types regarding expression of Lewis antigens. Furthermore we observed that sialylated compounds other than sialyl-Le(a) are responsible for histochemical detection of sialomucins and that sulpho-Le(a/c) is expressed in the presence or absence of sulphomucins detected using histochemistry. We conclude that mucin immunohistochemistry may replace classic histochemistry for the classification of IM into complete and incomplete types. The present study challenges the distinction of type-II from type-III IM since we did not observe major differences in the expression profile of mucins and Lewis type-1 carbohydrates. Finally, it seems necessary to evaluate the predictive value of IM according to the presence of specific sulphated carbohydrates (e.g. sulpho-Le(a/c)) rather than histochemically detected sulphomucins. PMID- 11889605 TI - Fast and sensitive immunodetection of carcinoma cells in sentinel nodes. AB - In a number of clinical situations, especially in the context of the recent sentinel node concept, lymph-node involvement has to be determined intraoperatively. Since serious and dependable decisions are to be made according to the result of this examination, the most reliable method for the detection of tumour cells should be applied. We and others have shown previously that routine histological examination underestimates lymph-node metastases, and that immunohistochemistry (IHC) significantly improves the accuracy of staging. However, IHC has so far been difficult to apply to the intraoperative examination of cryosections since it has required too much time. We have developed a novel modification of IHC for the rapid detection of metastases of carcinomas in cryosections from lymph nodes. It is based on a unique directly labelled cytokeratin antibody, immunofluorescence, and a specially devised staining solution. This one-step staining procedure can be performed within 10 min. At the same time, its sensitivity is very high. Single tumour cells can easily be detected, and background staining is very low. The high sensitivity could result in a markedly improved reliability of sentinel node-based decisions. PMID- 11889606 TI - Renal cell carcinoma with syncytial giant cell component. AB - We report a case of clear cell renal cell carcinoma in which a prominent multinucleated giant cell component was intermingled with clear, granular, and spindle cells. Histological, ultrastructural, cytometric, and cytogenetic features of giant cells were similar to those of mononucleated cells in the tumor, and therefore they were not from stromal or osteoclast derivation. These giant cells had homogeneous, finely granular, abundant cytoplasm, often with scalloped cell borders, and contained from 5 to more than 50 nuclei, all of them very similar in size and shape, with prominent central nucleoli. Occasionally, surrounding inflammatory cells were also engulfed in the cytoplasm. This syncytial appearance was more similar to that of some giant cell carcinomas from the lung than to the pleomorphic giant cells often encountered in high grade renal cell tumors. Although the patient is alive and free of disease 6 years after diagnosis, a longer follow-up will be required to assess the potential prognostic influence of this peculiar histological appearance. PMID- 11889612 TI - [Safety in anaesthesia--are there new concepts for patients with neuromuscular diseases?]. PMID- 11889607 TI - Primary leiomyosarcoma of the pancreas--a case report and case review. PMID- 11889608 TI - Neuromuscular and vascular hamartoma of the cecum. PMID- 11889613 TI - [Anesthesia in neuromuscular disorders. Part 2: specific disorders]. AB - The neuromuscular disorders described are divided into four groups: motoneuron diseases, peripheral neuropathies, disturbances of neuromuscular transmission and myopathies. In motoneuron diseases problems mainly result from respiratory insufficiency and the predisposition for aspiration caused by progressive muscular weakness. Depolarising muscle relaxants may elicit myotonic reaction and massive hyperkalemia. In contrast to non-depolarising muscle relaxants there may be an extreme hypersensitivity. In peripheral neuropathies the cardiac function is often limited whereby dysautonomia may enhance cardiovascular instability. The negative inotropic effect of anaesthetic agents must be observed with care and patients with higher degree of AV blocks may need a cardiac pacemaker during general anaesthesia. The Charcot-Marie-Tooth-Syndrome is characterized with a high sensitivity to thiopental. Disturbances of neuromuscular transmission frequently cause respiratory problems The fluctuating weakness of bulbar and respiratory muscles may impair swallowing and can lead to recurrent aspirations. Due to the reduced number of acetylcholine receptors the sensitivity to non depolarizing muscle relaxants is elevated and the response to succinylcholine is reduced. Drugs reducing neuromuscular transmission such as antibiotics and beta blockers may enhance these symptoms and should be avoided. In progressive muscular dystrophies the anaesthetic risk is mainly dependent on cardiac and respiratory impairment. Administration of succinylcholine leads to the risk of hyperkalmic cardiac arrest. Patients with metabolic myopathies are also at risk due to the involvement of cardiac muscle but respiratory problems are less frequent. Muscle metabolism should be supported by administration of substrates depending on the underlying disorder. In membrane disorders muscle rigidity (myotonic reactions) or weakness may lead to respiratory insufficiency. In addition to the depolarising muscle relaxants also anticholinesterase drugs, hypothermia and dyskalaemia can evoke myotonic reactions. PMID- 11889614 TI - [In-vitro-effects of cocaine in skeletal muscle specimens of patients susceptible to malignant hyperthermia]. AB - OBJECTIVE: The abuse of cocaine can cause serious medical complications like tachycardia, rhabdomyolysis, and hyperthermia. Because of the clinical similarities, it has been suggested that cocaine might be a trigger of malignant hyperthermia (MH). Therefore, aim of this study was to investigate the in-vitro effects of cocaine in skeletal muscle specimens of MH susceptible (MHS) and normal (MHN) patients. METHODS: 62 patients undergoing the in-vitro contracture test (IVCT) according to the protocol of the European MH Group (EMHG) for diagnosis of MH susceptibility were included in this study. In muscle specimens surplus to diagnostic requirements cocaine was added in order to achieve tissue bath concentrations of 0.01, 0.1 and 1.0 mM. The contracture development and twitch response have been registered. RESULTS: 21 patients were diagnosed as MHS and 36 patients as MHN. 5 patients tested as MH-equivocal (MHE) were excluded from the study. Following bolus administration of cocaine, no contracture development was observed in MHS, as well as MHN specimens. The muscle twitch decreased after cocaine administration significantly in both diagnostic groups. CONCLUSION: In contrast to the established MH trigger substances like volatile anaesthetics, cocaine produced no contracture development in MHS muscle specimens. Furthermore, cocaine produced a negative inotropic effect in all skeletal muscle preparations, which might be explained by local anaesthetic effects. Regarding these results, cocaine seems not to be a MH trigger agent. PMID- 11889615 TI - [Analgosedation, analgesia with remifentanil in incontinence surgery via tension free vaginal tape]. AB - OBJECTIVE: The tension-free vaginal tape operation (TVT) is a new surgical treatment of stress urinary incontinence in women. The tape has to be placed at the level of midurethra in the left and right paraurethral canal and has to be brought up to the abdominal wall in close contact with the back of the pubic bone with a special needle instrument. The performed anesthesia is decisive for the operative success, because a sufficient analgesia is demanded and, on the other hand, the correct placement and tension of the urethral tape has to be controlled by the patient performing a stress test by coughing and pressing. In this context the opioid remifentanil seems to be specially suitable because of its pharmacologic characteristics. METHODS: In this retrospective analysis the anesthesia related data of a total of 70 patients undergoing TVT surgery with remifentanil analgesia within one year were reviewed according to their anesthesia protocols. RESULTS: All patients underwent remifentanil analgesia in combination with local anesthesia. In no case the performed procedure had to be changed. In 7 cases a temporary decrease of the pulsoximetrically measured oxygen saturation to less-than-or-equal 92 % occurred, which could be treated by reduction of the infusion rate of remifentanil or by assisted ventilation. All patients were adequately able to perform the intraoperatively required stress test. 16 patients suffered from nausea and/or vomiting postoperatively. CONCLUSION: Continuous infusion of remifentanil is suitable for the short time profound analgesia needed for the TVT operation because of the pharmacologic characteristics of remifentanil. A antiemetic prophylaxis should be performed with this analgetic regime. PMID- 11889616 TI - [The visualisation of dura perforation and blood patches with ultrasound]. AB - The postdural puncture headache (PDPH) is a possible complication after spinal or epidural puncture. The therapeutic concept is usually organised step by step, but the epidural blood patch is the most reliable and effective therapy. In earlier studies myelographie, epidurographie and MRT were used, to visualise the localisation of the dural defect and to describe the effects of patching the epidural space. Our working group focused on the utilisation of ultrasound and we decided to use this technique for the visualisation of bloodpatches. With agreement of the local ethics committee we monitored the performance of 4 epidural bloodpatches in pregnant women, who suffered from PDPH. We used a General Electric LQ 400 ultrasonograph with a 7-MHz-probe. To ensure sterile conditions we used sterile ultrasound sleeves and sterile ultrasound gel. In 3 of 4 cases a continuity loss could be represented in the doppel layer signal of the dura. It had the size of the diameter of a Tuohy needle. In one case the defect of the dura was larger than in the preliminary investigations (2,5 - 3 mm). The ultrasonography of the epidural space was performed in the paramedian scan. All patches were placed by using the conventional loss of resistance technique with using online ultrasound support. The epidural puncture and the application of the blood patches were visualised simultaneously in all cases. With the injection of blood a brief expansion of the epidural space was seen. The patients recieved a mean injection volume of 17 ml sterile blood. 10 to 40 seconds after the injection of blood the disconituity of the dura doppel layer signal was no longer provable. Within a short time we could detect the increase of cerebrospinal fluid and the patients headache was treated sucessfully. The clinical use of this diagnostic technique can be found in the simultaneous presentation of the dura leakage and the intervention while performing an epidural bloodpatch. Since these informations are relevant for further clinical practice further investigations are warranted. PMID- 11889617 TI - [Sacroiliac joint blockage caused by bedding after spinal anaesthesia can simulate a neurological complication]. AB - Back pain after spinal anaesthesia is often attributed to the spinal anaesthesia and the accompanying puncture. We demonstrate two cases where this problem was not caused by spinal anaesthesia but was due to sacroiliac joint blockage caused by bedding. As far as we know, this complication has not been described yet. When taken into account, it can be treated easily and with success. PMID- 11889618 TI - [Decompressive craniectomy for severe intracranial hypertension due to cerebral infarction or meningoencephalitis]. AB - We describe the clinical course and outcome following decompressive craniectomy in six patients. Five patients suffered from severe intracranial hypertension due to middle cerebral artery infarction. In one patient the cause was bacterial meningoencephalitis. Acute clinical and neuroradiological signs of intracranial hypertension were seen in all cases. Following ineffective conventional brain edema therapy, decompressive craniectomy was undertaken. In five cases intracranial pressure was sufficiently lowered. One patient developed transtentorial herniation with subsequent brain death. Four patients with middle artery infarction showed moderate neurological disorders and one patient with bacterial meningoencephalitis recovered completely after treatment. Craniectomy in malignant middle artery infarction should be taken into consideration if conventional brain edema therapy does not sufficiently reduce critically raised intracranial pressure. Craniectomy provides development of brain herniation. This treatment may reduce high lethality rate and high frequency of severe neurological disorders. PMID- 11889619 TI - [Anaesthesia for patients suffering from mucoviscidosis]. PMID- 11889620 TI - [Up to now no effect of urodilatin on acute kidney failure in intensive care patients has been shown]. PMID- 11889621 TI - [Up to now no effect of urodilatin on acute kidney failure in intensive care patients has been shown. Reply]. PMID- 11889622 TI - Occurrence and therapy of space-occupying cystic lesions after brain tumor surgery. AB - OBJECTIVES: Space-occupying cystic lesions may develop in a variable time after resection of particular intracranial tumors, representing a small amount of complications of these procedures. We present our experience with the development and operative treatment of such postoperative cystic lesions in order to identify possible risk factors and to optimize the treatment. METHODS: The records and neuroradiological findings of patients, operated on either gliomas and meningeomas or craniopharyngeomas, who developed symptomatic cystic lesions in the former tumor resection area during the last ten years, were analyzed. RESULTS: 31 patients (2.5%) out of a total of 1240 corresponding tumor operations were identified. The mean age among the 20 female and 11 male patients was 47 years (12-74 years). In 17 patients (55%) the cystic lesion occured within 6 months after tumor resection (mean 5.6 weeks) and in 14 patients (45%) later than 6 months postoperatively (mean 3.6 years). 22 patients (71%) had malignant tumors and 16 patients (52%) had previous radiation therapy. 14 patients (45%) had more than one tumor resection at the same location and one patient had a postoperative meningitis as predisposing factor for the cyst-formation. All patients profited of the various definitive treatment modalities: repetitive percutaneous puncture/external drainage (5 patients), craniotomy for cyst-resection/ fenestration without (5 patients) and with Rickham-catheter implantation (10 patients), endoscopic cyst-fenestration with Rickham-catheter implantation (3 patients) and implantation of cysto-atrial or cysto-peritoneal shunts (8 patients). CONCLUSIONS: Symptomatic cystic lesions developing after brain tumor resection may occur as early - (5.6 weeks) or as late - (3.6 years) complications and though predisposing factors, like malignant primary tumor, preceding radiation therapy and multiple tumor resections can be identified, the reason for their occurrence remains unclear. A variety of effective therapy options is applicable but should consider the patients condition and prognosis. PMID- 11889623 TI - Schwannomas of the brachial plexus--diagnostic and surgical problems. AB - OBJECTIVE: This paper reports four cases of schwannomas of the brachial plexus operated upon with good outcome and discusses the CT and MRI findings and the best surgical treatment of these lesions. DESIGN: All patients had large schwannomas (more than 4 cm in diameter) presenting as painless masses in the supraclavear region, explored by CT and MRI. A homogeneous mass, hypo-isointense in T1- and hyperintense in T2-weighted images, with well-defined margins, is in favor of a schwannoma. The nerve of origin, external to the tumor mass, may be defined on MRI. INTERVENTION: All patients have been operated upon using microsurgical technique: enucleation of the tumor content, piecemal removal of the capsule, identification and preservation of the neural elements were the main goals of the operation in all cases. OUTCOME: Postoperatively, one patient experienced transient deficit of the deltoid muscle (two weeks). Actually, all four patients are symptom-free with no tumor recurrence, 6 months to 7 years after the operation. CONCLUSIONS: A correct preoperative diagnosis of schwannoma of the brachial plexus may be obtained by MRI, which shows a rather typical aspect; on the other hand, needle aspiration histology and open biopsies should be avoided. The microsurgical treatment with preservation of the neural structures, as for schwannomas of the cranial and spinal nerves, results in good outcome without recurrences. PMID- 11889625 TI - [Delayed traumatic CSF-fistulas: a retrospective analysis]. AB - Immediate posttraumatic CSF-fistulas are a well known entity after severe head injury. Delayed onset of rhinorrhea is considered to be rare. In the last 5 years 7 patients were treated in our department, who developed rhinorrhea 2-25 years after trauma. All patients went through episodes of meningitis. In 4 cases intermittent rhinorrhea was reported. In all cases a bony defect of the anterior skull base was detected by coronal bone window CT-scan. In three of them an encephalocele was revealed by MR-scanning. Treatment consisted in reconstruction of anterior skull base with a pedicled galeal-pericranial flap via a bifrontal craniotomy and went out without any complications. Delayed rhinorrhea after severe head injury is not a rare curiosity. In cases of bony defects after head injury reconstruction of anterior skull base is recommended to prevent episodes of recurrent meningitis. PMID- 11889624 TI - [Computer assisted plastic closure of calvarial defects]. AB - Large bony skull defects sometimes face problems of some pathophysiological effects, protection of the underlying brain, the impaired appearance, and from a psychological point of view. A computer-assisted method is presented, which has been successfully used in 44 patients. From 3-D-CT data a phantom was built in which Refobacin-Palacos was modeled for individual requirements. The plastic can be implanted after sterilization and leads to excellent cosmetical results. The use of a negative model reveals the possibility of an extended field of application without the necessity of highly specialized computersystems. PMID- 11889626 TI - [The posterior lumbar interbody fusion with cages (PLIF) and transpedicular stabilization]. AB - The development of intervertebral cages has significantly innovated the original technique of posterior lumbar interbody fusion (PLIF). In this study we present the results of patients treated for degenerative or postoperative segmental spinal instabilities by PLIF with cages and pedicular stabilisation (360 degrees instrumentation). Between 1992 and 1999 we implanted either CFRP-, PEEK- or Titanium-cages in 86 patients. 78 patients were adequately followed up over a period of at least 12 months (average 2,6 years). 5 patients were stabilised over 2 segments, so that ultimately 83 fused segments were evaluated.15% of all patients had an excellent, 51% a good, 28% a moderate and 5% an insufficient clinical result. Degenerative instabilities had a better outcome with 73% good or excellent clinical results, compared to postoperative instabilities (56%). Based on stringent radiographic fusion criteria we found true bony fusion in 52% of all segments after 12 months, 63% after 24 months, 72% after 36 months, and 78% after 48 months. In 21 segments cage packing was performed with autologous spongiosa, while in 62 segments a combination of cortical bone and spongiosa obtained from osseous structures at the operation-site were used as packing material. At the 24 month radiographic control we found a slightly higher fusion rate for those segments treated with autologous spongiosa obtained from the iliac crest. Neither for cages nor for pedicular screws was implant failure or material fatigue found. Serious entero-, pulmo-, cardio- or urological complications were not observed. Nonetheless the necessity for operative revision was 9%. A postoperative semiquantitative evaluation of segments neighbouring the fused vertebra revealed in 28% an increase in degenerative changes. Particularly after 360 degrees instrumentation, interpretation of the fusion-status should be based on structural and not on functional criteria. The modification of PLIF with cages compared to the use of only autologous spongiosa has the advantage of a high primary stability. Long-term studies are necessary to determine the implications of a radiographically evident uncertain fusion-status. PMID- 11889627 TI - [Patients' assessment of the importance of measures relevant for treatment in lumbar disc herniations and their satisfaction--a prospective study]. AB - In modern health system hospital is integral part of patients treatment. The hospital competes with an ubiquitous medical supply of adjacent institutions and general doctors. As a part of the medical supply chain a hospital may not exist as an "island" itself. Necessity in health politics and economics leads to image and advertisement strategies for a long term survival of hospitals. An analysis of medical output in patients treatment will build the basis for future development and strategies. By a questionnaire 170 patients suffering from a lumbar disc herniation were asked before starting medical treatment about their expectations and at the end of hospitalisation about their contentment with their stay in hospital. The very high expectations in medical treatment and nursing were fulfilled by the clinical staff. Here all patients were much more content they expected priorly. But the patients valued a distinct deficit in contentment in the co-treatment by other clinical faculties of the hospital. The reason of these results were explained by big distances inside the hospital and long time waiting between medical examinations. Our results show that intensive care by physicians and nurses increase contentment of patients. An additional improvement of patients judgement about hospital services will be possible by introduction of a routine co-treatment of other medical faculties. PMID- 11889629 TI - [Stapling of hemorrhoids - an alternative therapeutic option in the future as well?]. PMID- 11889630 TI - [Renaissance of the Franz Konig bilateral lip repair procedure - clinical experiences with an one hundred year old technique]. AB - Surgical correction of bilateral cleft lips is known to have a lot of problems. The surgical principles of treatment of bilateral cleft lips are similar to those of unilateral clefts but differ in the area of the prolabium due to specific anatomical disorders of the orbicularis oris muscle. The postoperative results of simultaneous bilateral cleft lip repair according to Konig were analysed retrospectively in 15 young children (6.1 +/- 1.1 years) paying special emphasis to the aesthetic and functional postoperative outcome of the upper lip and nose. The mean values were compared with measurements from normal infants at ages 8.3 +/- 1.8 years. Lip height and lip length were in 87 % similar to those of the age matched normal group. Only two cleft patients showed a slightly shorter lip. Distortions of the lip function were not obvious. Our data show that Konig's surgical procedure of bilateral cleft lip closure meets the requirements of modern surgical concepts of cleft lip repair and should belong to the armamentarium of modern face surgery. PMID- 11889631 TI - [Results two years after stapler hemorrhoidectomy versus Milligan-Morgan procedure]. AB - The aim of this investigation was to demonstrate possible advantages of stapler hemorrhoidectomy in comparison to the Milligan-Morgan procedure. 96 patients with an average age of 54 years were treated in a two year period (7/1998-8/2000) by stapler hemorrhoidectomy. The complication rate was 12.5 % and included one mechanical stapler defect, two cases of bleeding, five cases of urinary retention and four of perianal edema. The use of analgesics was small with 70 % requiring no medication at all. Hospitalisation post-operatively was 3.3 days with patients under 65 years old and 4.7 days in those over 65 years. These data were compared retrospectively to that of Milligan-Morgan hemorrhoidectomy (214 patients) performed between January 1990-December 1997. The stapler patients had less pain, fewer complications and shorter hospitalisation. Using a questionnaire, all stapler patients and 50 Milligan-Morgan controls were evaluated. 78 % responded at 13.8 months after stapler hemorrhoidectomy, 63 % 54.1 months after Milligan Morgan. The degree of satisfaction was high in both groups (93 vs 94 %). One patient in the Milligan-Morgan group suffered a recurrence. No further symptoms had been experienced by 57 % after stapler, 68 % after Milligan-Morgan procedure. Faecal continence represents a problem in the stapler group. Stapler hemorrhoidectomy is an effective treatment for IIIrd degree hemorrhoids. In comparison to the Milligan-Morgan procedure, it has advantages in the early post operative period. Defecation problems can occur with an unknown prognosis. Without long-term results and because of the comparatively high cost of the procedure the indication for stapler hemorrhoidectomy should be carefully made. PMID- 11889632 TI - [Stapler hemorrhoidectomy versus conventional procedures - a clinical study]. AB - Stapled Hemorrhoidectomy is when correctly indicated an easy feasible operative procedure for prolapsing internal hemorrhoids with or without a mucosal prolapse offering benefits to the patient. From July 1998 to October 2000 we treated 152 patients with a mean age of 52 (24-91) years for hemorrhoids within this study. We compared 72 patients, treated with stapled hemorrhoidectomy according to Koblandin-Longo with 80 patients who underwent a "conventional" reconstructive operation (Parks or Fansler-Arnold). All resected material was histopathologically examined. With stapler hemorrhoidectomy we found on average shorter operation times (22 vs. 53 min, p < 0.01), shorter hospitalisation (3 vs. 6.1 d, p < 0.01), significantly less postoperative pain (VAS 0-10: 1.83 vs. 3.70, p < 0.01) and fewer cumulative requests for analgesia by the patients (0.92 vs. 3.11 single doses, p < 0.01). The complication rate was 4 % in the stapler group and 11 % in the conventional group. Stapled hemorrhoidectomy was carried out only in patients with 3 degrees hemorrhoids with or without mucosal prolapse. The conventional group consisted of patients with 3 degrees prolapsing or 4 degrees fixated external hemorrhoids. Although very promising results are actually described with stapler hemorrhoidectomy, the established conventional reconstructive operations should be continued until long-term results are published. PMID- 11889633 TI - [Evaluation of Longo's technique for haemorrhoidectomy by doppler ultrasound measurement of the superior rectal artery]. AB - Stapler-haemorrhoidectomy causes theoretically a durable reposition of the prolapsed haemorrhoidal cushions and a reduction of the arterial inflow by clipping mucosa and submucosa. Until now, however, no exact data exist with respect to a potential reduction of the arterial inflow. METHODS: The question of a sufficient interruption of the end branches of the superior rectal artery should be answered with doppler ultrasound measurements before and after stapler haemorrhoidectomy. RESULTS: The measurements were performed on 45 patients before and one month after stapler-haemorrhoidectomy. Preoperatively in all patients the three main branches of the artery at three, seven and eleven o'clock could be detected by doppler ultrasound. In 67 % of the patients a fourth, in 16 % a fifth and in 13 % a sixth vessel could be located. One month postoperatively in 80 % of the patients all main branches were further seen. In 16 % of the cases two main vessels, in 4 % only one main vessel could be identified. There was no correlation between postoperative outcome and number of vessels detected postoperatively. CONCLUSION: It is concluded that the postoperative outcome after stapler-haemorrhoidectomy does not depend on the complete interruption of the arterial inflow of the haemorrhoids. The complete reposition of the haemorrhoidal prolapse and thereby the improvement of the venous reflux out of the haemorrhoidal cushions might be more important. PMID- 11889634 TI - [Anal pressures after stapler hemorrhoidectomy - a prospective analysis of 33 patients]. AB - PURPOSE: Determination of the effects of staplerhemorrhoidectomy as a new method of surgery, type of staplers and way of anal retraction on anal pressures. PATIENTS AND METHODS: In 33 patients (mean age 56 ys.) with third degree hemorrhoids who underwent staplerhemorrhoidectomy in the Marienhospital Gelsenkirchen between 1998 and 1999, anal resting and squeezing pressures were measured before and after the operation. On an average the postoperative examination was performed 47 days after operation. RESULTS: Anal resting pressures decreased significantly from 69 (23) to 58 (18) mmHg (p < 0.01) in contrast to the anal squeezing pressures (171 (60) and 170 (58) mmHg). There was a relatively greater decrease in anal resting pressure using a Parks' retractor in comparison to the use of a vaginal speculum. The decrease of resting pressure did not depend on the type of stapler used (Ethicon(c) SDH 33, n = 14 and Autosuture(c) CEEA 31, n = 19). CONCLUSION: Hemorrhoidectomy using a circular stapler leads to a significant reduction of the anal resting pressure, whereas squeezing pressures remain constant. The reduction is more pronounced if a Parks' retractor is used. PMID- 11889635 TI - [Anal sphincter-CT and dorsal sphincteropexy - a new approach in therapy of obstructive defecation disorder]. AB - The dysplasia of anal sphincter represents an obstructive defecation disorder. The disease is known as "anterior displaced anus" in pediatric surgery. An anorectal malformation with missing dorsal osseous fixation of the sphincter complex is the underlying cause. Beyond clinical symptoms and examination result (anterior displaced anus with palpable dorsal gap) the defect can be visualized by computered tomography. A surgical correction is possible by the simple intervention of dorsal sphincteropexy. In our trial with 48 patients (male n = 12, female n = 36, mean age 51 +/- 17 years, follow-up in 39 patients) a significant improvement of defecation could be achieved in 46 % of the patients. In correlation to a good clinical outcome a significant reduction in the defecation score was observed. 10 % of the patients had only small changes in symptoms. However, the proportion of dissatisfied patients was relatively high with 44 %. In this group patients with long-standing chronic constipation and laxative abusus were found more often and the rate of previous anal or abdominal surgery was quite higher. Dissatisfied patients showed a higher variation in symptoms of pelvic floor disorders (e. g. anal pain syndrome) besides the rectal evacuation disorder. In addition to the heterogenity of symptoms chronic alterations of pelvic floor structures might create worse results in patients with chronic constipation. In spite of a lot of publications dealing with the functional anatomy of the pelvic floor only a few investigations on the dorsal sphincter dysplasia in patients with rectal evacuation disorder are found in the literature. Further investigations on this disorder are necessary. PMID- 11889636 TI - [Quality of life assessment after surgery for diverticulitis - a follow-up study]. AB - BACKGROUND: Laparoscopic sigmoid resection is a well established procedure for surgical treatment of benign colorectal diseases. The aim of the present study was to assess the longterm quality of life of patients who underwent laparoscopic sigmoid resection for sigmoid diverticulitis. Differences in health related life quality to the open-conventional approach were evaluated in a matched pair analysis (age, gender, Hinchey-Stage, Type of Surgery) using a validated quality of life instrument. METHODS: A total of 45 matched pairs (laparoscopic/open) operated for diverticulitis at stage I-IIa (Hinchey classification) were included in this study. The quality of life was measured with the Short-Form-36-Health Survey (SF-36), a standardized questionnaire with 8 scales and 36 items. The follow-up period was at least 2 years (mean 62.2 months). RESULTS: Pair members (n = 45) operated via laparoscopic or open approach for Hinchey I-IIa diverticulitis were of the same sex (21 female/24 male pairs) and age at time of surgery (range: lap.: 53.5-66 years; open: 53.5-67 years). Mean follow-up periods for patients operated laparoscopically and with open procedure were 2 (range: 1 3) and 7 (range: 5-9) years, respectively. The SF-36 scale scores for both groups appeared high and only slightly below a validated norm population. This represents a high quality of life after open as well as laparoscopic surgery for sigmadiverticulitis. No significant differences were apparent between the 45 matched-pairs. Pairs 65 years old or older presented no significantly different score values compared to those younger than 65 years. CONCLUSIONS: The long-term follow-up data in this age and sex matched pair analysis showed favorable results after open as well as laparoscopic surgery for sigmadiverticulitis. No statistically significant differences were observed between the two surgical techniques. Self-reports by the patient concerning his or her health condition, recovery and quality of life following any surgical procedure are needed to assess valid outcome data of new surgical treatments including a critical evaluation of all its benefits and burdens. PMID- 11889637 TI - [Therapeutic results after papilloduodenal injuries following ERCP]. AB - INTRODUCTION: Our goal was to compare operative vs. conservative therapeutic strategies after injuries following ERCP. METHODS: Eight patients with ERCP induced injuries were surveyed retrospectively. Four of them were treated operatively, four conservatively. Criteria for an operative therapy were clinical and radiological findings and laboratory data. RESULTS: The four patients that were treated conservatively had an uncomplicated course whereas three of four patients treated operatively had long and complicated stays. In these patients the operation was performed more than 24 hours after injury. All of them showed advanced biliary peritonitis. One patient was operated on within 24 hours. He was discharged after a short stay without complications. All injuries were located in the retroperitoneum. Five patients showed anatomical abnormality of either duodenum, papilla or common bile duct. In five cases the duodenum was involved in the injury. CONCLUSIONS: The course of disease of the operated patients was longer and more complicated compared to those treated conservatively. According to our data the timing of the operation seems to be an important criterion with respect to the prognosis. Due to the small number of patients, whether conservative therapy should be preferred cannot be determined. The role of the location of injury is also not clarified. PMID- 11889638 TI - [Prospective study on patients outcome following laparoscopic vs. open cholecystectomy]. AB - AIMS OF THE STUDY: Differences in outcome between patients undergoing laparoscopic (LC) vs. open cholecystectomy (OC) should be examined under objective and subjective aspects. METHODS: We prospectively evaluated the postoperative course of 135 patients who underwent LC or OC in 1999. In the first step we examined the recover period with the help of the modified McPeek-Index. In the second phase during a 35 postoperative days-spanned analysis all patients noted there physical, emotional and social well-being in a circulating standard questionnaire, based on the modified Gastrointestinal Quality Life Index (GIQLI). RESULTS: Responses were obtained from 103 patients (76.3 %) undergoing 29 OC and 74 LC. 21.4 % of the patients aged 70 and older or had perioperative risks > II in ASA-Classification (LC 19.8 vs. OC 30.1 %). The (objective) McPeek-Outcome was similar in both groups, with no statistical advantage for LC (best score: 69 % LC vs. 62 % OC). The subjective assessment of the patients showed that patients having LC felt fully fit 10.2 days earlier than patients after OC (23.9 vs. 34.1 days). Patients in the LC-group returned to work after an average of 24.7 days, compared with 42.2 days following OC. The main finding of the postal questionnaire was a significantly earlier recruitment in physical, emotional and social status following LC in the group of aged > 70 and > ASA II-Score-patients, in contrast to control-OC-group. CONCLUSION: The study suggests an additional advantage in surgical outcome after LC, in comparison to OC. The laparoscopic approach is the preferable procedure to treat especially older and comorbide patients, when local or anesthesiological contraindications are absent. PMID- 11889639 TI - [Long-term results of benign bile duct strictures after treatment with pedicled jejunal patches]. AB - Benign strictures of the common bile duct after surgery or due to gallstones may lead to obstruction and derangement of bile drainage in the extrahepatic biliary system. Although the treatment of choice in these situations is the endoscopic dilatation, in some cases with stenosis of a long segment of the bile duct a partial replacement with a vascularised jejunal patch may be possible and useful. To our knowledge, there are no reports on long-term results of the procedure. We describe the course, the surgical technique and long-term results of four patients with a jejunal patch reconstruction of the common bile duct. Ten years after surgery there were no radiologic or laboratory signs of a restenosis of the common bile duct. PMID- 11889640 TI - [Benign peptic stricture of the middle third of the esophagus]. AB - By means of a case report classification, etiology and pathogenesis of benign esophageal strictures are discussed. In the presented case an endobrachyesophagus with peptic stricture of the middle third of the esophagus was found. The most common conservative treatment consists of esophageal bouginage combined with long term medication of proton pump inhibitors as was also done in this case. Depending on the physical status of the patient surgical treatment is given if repeated bouginage of the stricture during at least 3 months becomes necessary. Resection of the stricture or antireflux surgery in combination with bouginage of the stricture are current surgical options. PMID- 11889642 TI - [Infected urachal cyst in an adult - a case report and review of literature]. AB - Infected urachal cysts are a rare clinical manifestation in adults. We present the case of a female patient with an infected urachal cyst, discuss the embryology, clinical presentation, diagnostics and the therapeutic procedure and make a comparison with the literature. PMID- 11889643 TI - [Pierre-Marie-Bamberger syndrome - a paraneoplastic syndrome of lung cancer - a case report]. AB - The Pierre-Marie-Bamberger syndrome is a rare paraneoplastic syndrome caused by bronchial carcinoma. Typical signs are symmetric periostoses on the diaphyses of the long tubular bones, clubbed fingers and toes with eye-glass shape of the nails, neuro-vegetative disturbances and dysproteinemia. We report a 37-year-old patient with long-term nicotine abusus, who attracted attention by symptoms of a Pierre-Marie-Bamberger syndrome. Further diagnostics revealed a tumor in the apex of the left lung. After lobectomy of the upper lobe of the left lung the symptoms are completly disappeared. PMID- 11889644 TI - [On the history of surgical treatments of peritonitis]. AB - The surgical treatment of peritonitis started with the first laparotomy for an infected ovarian cyst by McDowell in the beginning of the 19(th) century. Thereafter the surgical approach developed parallel to the advances in abdominal surgery. In the last decade of the 19(th) century Mikulicz felt that laparotomy was indicated in all patients with purulent peritonitis. In the beginning of the 20(th) century Korte and Kirschner defined the principles of surgery for peritonitis that are valid up to this day: early surgical intervention, elimination of the source of infection, and peritoneal toilet. Since that time surgeons have discussed the utility of draining and irrigating the peritoneal cavity. Postoperative lavage was already advocated in the beginning of the last century, but generally regarded ineffective. Thirty years ago postoperative lavage was again strongly advocated, but evidence for its benefit is still missing. The history of the treatment of peritonitis examplifies that basic concepts which have been discovered in the past often remain valid and can serve as guidance for a long time. Such discoveries deserve recognition and should give reason for a critical appraisal of current practice. Thus, the statement of Trendelenburg made one hundred years ago remains true: "...in medicine, too, the today is based on the yesterday, and to follow a gradual development is of immense interest. In daily practice, especially in surgery and obstetrics, many things are much older than an ignorant can imagine." PMID- 11889645 TI - Factors predicting orofacial pain patient satisfaction with improvement. AB - AIMS: To determine psychosocial predictors of patients' ratings of satisfaction with improvement and subjective pain relief. This study also examined the underlying components of patient satisfaction with improvement, as assessed at follow-up. METHODS: The sample consisted of 107 chronic orofacial pain patients evaluated at a university-based orofacial pain clinic and referred for treatment with individualized treatment plans. Pain and psychosocial functioning were assessed with standard, reliable, validated self-report instruments administered at the initial evaluation. Follow-up data were collected via a telephone administered structured interview 8 months after the initial evaluation. Regression methodology was used to determine prediction models for satisfaction with improvement and subjective pain relief. Patient ratings of the quality of the caregiver communication were used as a control variable in all analyses. RESULTS: Quality of caregiver communication predicted approximately 10 to 14% of the variance in outcomes in all models. Greater initial use of cognitive coping strategies and reduced depression predicted higher ratings of satisfaction with improvement and increased pain relief. When concurrent relationships among variables at the follow-up were examined, greater subjective pain relief since the evaluation, lower current pain, and higher ratings of overall mood were significant predictors of patient satisfaction with improvement. CONCLUSION: This study is one of the first to report that the use of certain cognitive coping strategies is associated with positive outcome for patients suffering from orofacial pain. These findings underscore the importance of individual differences on behavioral and psychosocial parameters in the prediction of patients' subjective evaluation of treatment outcome. PMID- 11889646 TI - Diffusion model of pain language and quality of life in orofacial pain patients. AB - AIMS: To address the following questions: (1) Which words are preferred by different groups of orofacial pain patients to describe their pain experience? (2) Is it possible, based on such descriptions, to obtain a clinical differential diagnosis in these patients? (3) Is there any relationship between the verbal description of pain and self-rated quality of life (QOL)? (4) Can a pattern of modulation of pain language by affective variables (diffusion model) be recognized in orofacial pain patients, as it has in other chronic pain patients? and (5) If so, what might be the clinical usefulness of assessing pain language in these patients? METHODS: A total of 332 consecutive orofacial pain patients filled out an Italian Pain Questionnaire (the Italian analog of the McGill Pain Questionnaire) and were then divided into 6 diagnostic subgroups (sample 1) based on history and clinical findings. In a double-blind setting, the distribution of pain descriptors and indexes was statistically evaluated. From sample 1, a randomly selected sample of 121 patients (sample 2) also filled out a QOL categorical scale. The results of both tests in this sample were compared statistically. RESULTS: Some significant differences among diagnostic subgroups were found for choice of descriptors and for pain intensity. When a patient's pain description was compared to the corresponding self-evaluation of QOL, a self perceived worsening of QOL revealed a good correlation with an increase in the number of words chosen, pain intensity, and affective and sensory pain descriptors. A similar significant association was found between self-assessed anxiety and/or depression and the same items. CONCLUSION: Although trends in patients' choice of descriptors were evident, differential diagnosis based on only a pain questionnaire was not possible in the different groups of orofacial pain patients examined in this study. The present study suggests the presence of a phenomenon of diffusion in the language of those patients who were experiencing a worsening of their QOL as a result of pain and consequent psychologic distress. This observation can be of clinical usefulness by enhancing the sensitivity of the clinician to the suffering and affective distress experienced by the patient, and it also can be helpful in refining the therapeutic approach for each individual patient. PMID- 11889647 TI - Physical self-regulation training for the management of temporomandibular disorders. AB - AIMS: To evaluate the long-term effectiveness of a brief skills training program for the management of chronic facial muscle pain. This program of physical self regulation (PSR) involved primarily training in breathing, postural relaxation, and proprioceptive re-education. METHODS: Physical self-regulation training was presented by a dentist during two 50-minute sessions spaced at 3-week intervals and was compared to a standard dental care (SDC) program that included a flat plane intraoral appliance and self-care instructions provided by a dentist. Participants (n = 44) were initially evaluated by a dentist experienced in the diagnosis and management of orofacial pain and were determined to have myofascial pain (Type 1a and 1b diagnoses per the Research Diagnostic Criteria) prior to random assignment to either the PSR or SDC conditions. Posttreatment evaluations 6 weeks and 26 weeks after treatment had begun were conducted by a dentist who was not aware of which treatment the participants received. RESULTS: Initial results indicated that pain severity and life interference from pain were reduced in both groups (P < 0.001), while perception of control was increased (P < 0.001), as was incisal opening without pain (P < 0.05). At the 26-week follow-up, the PSR group reported less pain (P < 0.04) and greater incisal opening, both with (P < 0.04) and without (P < 0.01) pain, than the SDC group. There were also significant decreases (P < 0.05) in affective distress, somatization, obsessive compulsive symptoms, tender point sensitivity, awareness of tooth contact, and sleep dysfunction for both groups over time. CONCLUSION: The findings support the use of PSR for the short- and long-term management of muscle pain in the facial region. These results are discussed in terms of the potential mechanisms by which self-regulation treatment strategies are effective for the management of these pain disorders. PMID- 11889648 TI - New insights into peripheral chemical mediators of temporomandibular pain and inflammation. PMID- 11889649 TI - Oral motor parafunctions among heavy drug addicts and their effects on signs and symptoms of temporomandibular disorders. AB - AIMS: To investigate the prevalence of temporomandibular disorders (TMD), bruxism, and other oral habits among drug addicts compared to a normal, non addicted, matched control population, and to assess the detrimental effect of long-term drug abuse on the parameters studied. METHODS: Subjects included 55 drug-addicted patients (51 males and 4 females) randomly selected from long-term addicts using "hard" narcotics and attending a methadone maintenance center and a control group of 52 normal non-addicted individuals (48 males and 4 females) matched to the addicts for age, gender, and socioeconomic status. A clinical examination and a questionnaire were used. One examiner determined that all questions were correctly understood and answered, and a second examiner performed the clinical examinations and was unaware of the results of the questionnaire. RESULTS: The addicted group had a high prevalence of orofacial motor behavior (bruxing, clenching) as well as signs and symptoms of TMD (morning headache, joint noises, joint and masticatory muscle tenderness to palpation, and tooth wear) compared to the controls. Active (voluntary) jaw opening was significantly smaller, although within an acceptable range when compared to the controls. CONCLUSION: Long-term drug abuse detrimentally affects the stomatognathic system, as expressed in a high prevalence of oral motor behavior and signs and symptoms of TMD. PMID- 11889650 TI - Lack of associations between occlusal and cephalometric measures, side imbalance in striatal D2 receptor binding, and sleep-related oromotor activities. AB - AIMS: First, to evaluate possible orofacial morphologic differences between sleep bruxers and non-bruxers, and second, to determine possible correlations between morphologic factors and striatal D2 receptor expression in persons with sleep related oromotor activities. METHODS: Twenty subjects were included in this study; half of them had polysomnographically confirmed oromotor values above the cutoff points for sleep bruxism. For all participants, 26 standard occlusal measures were recorded clinically and from dental study casts. In addition, 25 standard angular and linear measures were taken from standardized cephalometric films, and variables were derived to evaluate dental and skeletal relationships. Fourteen of the 20 participants had also participated in a previous study that included iodine-123-iodobenzamide (I-123-IBZM) and single-photon emission computed tomography (SPECT). For them, the side-to-side difference in striatal D2 receptor binding was determined as the neurochemical outcome measure. RESULTS: Following the classical Bonferroni adjustment for multiple testing, no morphologic differences were found between the sleep bruxers and the non-bruxers. In addition, none of the morphologic variables were significantly associated with the neuroimaging data. CONCLUSION: Taking into account the low power of this retrospective, exploratory study, the results suggest that the orofacial morphology of sleep bruxers does not differ from that of non-bruxers. In addition, morphologic factors are probably not involved in the asymmetry in striatal D2 receptor distribution that was previously observed in association with sleep bruxism. PMID- 11889651 TI - Spring Pain Conference. May 7-12, 2000. Grand Cayman Island. PMID- 11889652 TI - Neuroendocrine, immune, and local responses related to temporomandibular disorders. AB - Orofacial pain frequently originates from pathologic conditions in the masticatory muscles or temporomandibular joints (TMJs). The mediators and mechanisms that monitor pain and inflammation, centrally or peripherally, are of great interest in the search for new treatment modalities. The neuropeptides substance P (SP), calcitonin gene-related peptide (CGRP), and neuropeptide Y (NPY) have all been found at high levels in the synovial fluid of arthritic TMJs in association with spontaneous pain, while serotonin (5-HT) has been found in association with hyperalgesia/allodynia of the TMJ. Interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) have been found in arthritic TMJs, but not in healthy TMJs, in association with hyperalgesia/allodynia of the TMJ as well as spontaneous pain. Anterior open bite, which may be a clinical sign of TMJ destruction, has been found in association with high levels of CGRP, NPY, and IL-1 beta in the synovial fluid of the TMJ. Interleukin-1 beta has also been related to radiographic signs of joint destruction. Prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) are both present in the arthritic TMJ, and PGE2 has been shown to be associated with hyperalgesia/allodynia of the TMJ. Very little is known about pain and inflammatory mediators in muscles. However, we know that 5 HT and PGE2 are involved in the development of pain and hyperalgesia/allodynia of the masseter muscle in patients with fibromyalgia, whereas local myalgia (myofascial pain) seems to be modulated by other, as yet unknown mediators. Interaction between the peripheral nervous system (sensory and sympathetic nerves), the immune system, and local cells is probably of great importance for the modulation of pain and inflammation in the TMJ and orofacial musculature. PMID- 11889653 TI - Osteomyelitis in the diabetic patient: diagnosis and treatment. Part 1: Overview, diagnosis, and microbiology. AB - Foot ulcerations are common among people with diabetes and often lead to mixed infections that require hospitalization and create significant challenges for clinicians. Many result in contiguous bone infections. Regimens used to treat osteomyelitis often are seen as controversial. A subtle balance between medical and surgical therapy is necessary if a potentially curative outcome is to be achieved. The following article is one of a two-part series. This, the first manuscript, discusses the diagnosis and microbiology of contiguous osteomyelitis in the diabetic foot. PMID- 11889654 TI - Quantitative swab culture versus tissue biopsy: a comparison in chronic wounds. AB - Soft tissue infection present a significant obstacle to the healing of chronic wounds. Historically, the gold standard for determining wound bacterial bioburden has been the quantitative tissue biopsy. Nevertheless, tissue biopsies are not universally used in today's healthcare setting. Likely reasons include damage to healing tissue, the lack of facilities to process tissue biopsies, significant pain in sensate soft tissue, and increased expense with this modality. More recently, quantitative tissue swab culture has been suggested as a means to determine the wound bioburden. The authors prospectively studied 38 patients with chronic wounds of various etiologies to evaluate the correlation between quantitative wound biopsy and swab culture. Of the 38 biopsies performed, 74% indicated infection. Simultaneous swab culture of these 28 biopsies indicated infection in 22 of the 28 cases for a correlation of 79%. The authors concluded that quantitative swab culture provides a valuable adjunct in the management of chronic wounds. PMID- 11889655 TI - A tool to assess clinical signs and symptoms of localized infection in chronic wounds: development and reliability. AB - This paper reports on the development and testing of a tool designed to assess chronic wounds for the clinical signs and symptoms of localized infection. Thirty one wounds were assessed by two independent nurse observers for the signs and symptoms of infection using the Clinical Signs and Symptoms Checklist. The Clinical Signs and Symptoms Checklist delineates 12 signs and symptoms of infection (i.e., pain, erythema, edema, heat, purulent exudate, serous exudate with concurrent inflammation, delayed healing, discoloration of granulation tissue, friable granulation tissue, pocketing at the base of the wound, foul odor, and wound breakdown) and their definitions. The reliability of each sign or symptom on the checklist was calculated using percent agreement and the Kappa statistic. Percent agreement ranged from 65% to 100%, and Kappa statistics ranged from 0.53 to 1.00, excluding pocketing of the wound base. The reliability estimates obtained for signs and symptoms on the Clinical Signs and Symptoms Checklist compare favorably with other data regarding interclinician agreement on wound assessment. Incorporating a structured approach to assess and monitor for wound infection, such as the Clinical Signs and Symptoms Checklist, may improve clinician skill and accuracy in identifying this condition. PMID- 11889656 TI - Experimental chewing in myofascial pain patients. AB - AIMS: To evaluate the potential capacity of a chewing exercise to differentiate chronic myofascial pain (MFP) patients from healthy controls and to test whether there are distinct pain response differences among MFP patients. METHODS: Eighty nine subjects participated in the study; 49 were diagnosed as belonging to the MFP subgroup of temporomandibular disorders (TMD) and had suffered from MFP for at least 6 months, and 40 healthy age- and gender-matched subjects comprised the control group. After completion of a clinical examination, all subjects performed a chewing exercise. Subjects chewed on half a leaf of green casting wax for 9 minutes and then held their jaw at rest for another 9 minutes. They indicated the intensity of the pain experience on a visual analog scale (VAS) every 3 minutes from the beginning (P0) to the end (P18) of the chewing exercise. Only changes in pain report of more than 5 mm on the VAS were considered. Analysis of covariance with repeated measures was used to analyze fluctuations in pain levels during the test, with the pain level at baseline (P0) as a covariant. RESULTS: Statistical analysis revealed a significant main effect for group (MFP versus control); a significant main effect for activity (chewing versus rest); an interaction between activity and time; and an interaction between activity, time, and group. The latter revealed the significant effect of the chewing activity on pain levels in both groups along the axis of time and its recovery at rest. In the MFP patients, pain had increased by 32 mm at P9 in 84% of the patients and recovered to almost the initial pain levels by P18; 6% reported a decrease in pain sensation and 10% reported no change in pain. In the controls, pain had increased 4.9 mm by P9, a value within the recording error range of the scale. CONCLUSION: (1) A strenuous chewing exercise is a potentially beneficial tool in the diagnostic process of myofascial pain patients and, if validated, could be incorporated into clinical examinations. (2) The increase in pain intensity following the chewing exercise is typical of most of the MFP group. (3) The phenomenon of pain decrease in a small percentage of MFP patients should be further investigated. PMID- 11889657 TI - The mediolateral temporomandibular joint disc position: an in vivo quantitative study. AB - AIMS: Temporomandibular joint (TMJ) discs displaced simultaneously, dorsoventrally, and mediolaterally are assumed to be rotated. However, a pilot study performed with individualized oblique-axial scans on supposedly rotated discs did not show disc rotation consistently. The aim of this study was the quantitative evaluation of disc rotation on a larger sample size, assessing the mediolateral disc geometry and position by the use of a reference system determined by the condylar long axis. METHODS: Eighty-five TMJs from 50 subjects were analyzed. One series of sagittal and 1 of 14 individualized oblique-axial magnetic resonance (MR) scans were taken for each joint. The dorsoventral disc position was diagnosed by means of the sagittal scans. The mediolateral disc width and position were then measured on every oblique-axial scan. The width and midline was computed for each disc and its deviation from the perpendicular to the condylar long axis was calculated. Finally, a statistical analysis was performed to study whether the disc width and the direction of the disc midline varied between discs normally positioned and anteriorly displaced. RESULTS: The disc width varied significantly more within the anteriorly displaced discs than within the normal ones. The midline of the anteriorly displaced discs deviated more from the perpendicular to the condylar long axis than that of normally positioned discs and was mostly in a lateral direction. The disc midline also deviated more in the ventral than in the dorsal part of the disc. CONCLUSION: Most anteriorly displaced discs were laterally displaced and showed a larger width variation than normally positioned discs. This fact seems to indicate disc deformation. PMID- 11889658 TI - Dietary fiber intake in patients with myofascial face pain. AB - AIMS: To determine the impact of myofascial face pain (MFP) on dietary intake of selected nutrients. METHODS: Sixty-one MFP women meeting the criteria for the myofascial subtype of temporomandibular disorders completed a 4-day daily food intake diary, as well as self-report of pain severity, pain interference with eating, and depressive symptomatology. Nutrient intake for the MFP women was compared with a demographically-equivalent sample of community women participating in the federally-sponsored Continuing Survey of Food Intakes by Individuals (CFSII). Within the MFP sample, multiple linear regression analysis was used to test whether dietary fiber intake reduction was most likely due to pain adaptation, or to depressive symptomatology or associated appetite reduction. RESULTS: Only the subgroup of MFP patients with above-average pain severity showed reduced dietary fiber intake compared with the community sample. MFP patients did not differ from the community sample on other nutrient intake measures (i.e., total calories, protein, fat, carbohydrates and dietary fiber, calcium, and iron). Within the MFP sample, pain severity was significantly associated with reduced dietary fiber intake. This relationship persisted, after controlling for depressive symptomatology, appetite, and total calories. CONCLUSION: Myofascial face pain patients with more severe pain intensity are likely to reduce their intake of dietary fiber. This is likely due to an effort to decrease masticatory activity to avoid exacerbating facial pain. Since low dietary fiber, especially in combination with commonly prescribed medications for MFP, increases the risk of constipation and may exacerbate comorbid medical conditions, clinicians should recommend alternative dietary fiber sources for MFP patients. PMID- 11889659 TI - A randomized clinical trial using research diagnostic criteria for temporomandibular disorders-axis II to target clinic cases for a tailored self care TMD treatment program. AB - AIMS: To carry out a randomized clinical trial (RCT) contrasting usual conservative treatment of TMD by clinical TMD specialists with a structured self care intervention, targeted to clinic cases independent of TMD physical diagnosis, who were reporting minimal levels of psychosocial dysfunction; the intervention was delivered by dental hygienists in lieu of usual treatment. METHODS: The Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) was used to target subjects who exhibited minimal TMD-related psychosocial interference. Criteria for study inclusion were: (1) self-report of facial and/or masticatory muscle pain discomfort for which usual care was prescribed by the clinic TMD specialist; (2) RDC/TMD Axis II graded scale of chronic pain (GCP) score of 0, I, or II-Low. (3) Age 18 to 70 years. RESULTS: On 1-year follow-up, while both groups showed improvement in all clinical and self report categories measured, patients in the tailored self-care treatment program compared to usual TMD treatment showed significantly; (a) decreased TMD pain, (b) decreased pain-related interference in activity; (c) reduced number of masticatory muscles painful; (d) fewer additional visits for TMD treatment. Groups were comparable with regard to measures of vertical range of motion. The self-care program was associated with consistent, but non-statistically significant, trends towards lower levels of depression and somatization. Ability to cope with TMD, knowledge concerning TMD and patient satisfaction was significantly enhanced for the self-care group. No participating patients experienced physical or personal adverse effects during the 1-year post-treatment follow-up period. CONCLUSION: Use of RDC/TMD psychosocial assessment criteria can contribute to successful clinical decision-making for the management of TMD. PMID- 11889660 TI - The evolution of a scientific journal. PMID- 11889661 TI - The effectiveness of adding pharmacologic treatment with clonazepam or cyclobenzaprine to patient education and self-care for the treatment of jaw pain upon awakening: a randomized clinical trial. AB - AIMS: To compare the relative effectiveness of a benzodiazepine (clonazepam), a muscle relaxant (cyclobenzaprine), and a placebo for the treatment of jaw pain upon awakening, when each is combined with the recommended nonpharmacological components of initial medical management. METHODS: Forty-one subjects were recruited with a diagnosis of myofascial pain based on the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). All subjects were given education about TMD and a self-care program. Subjects were randomized into 1 of 3 groups: clonazepam (0.5 mg/night), cyclobenzaprine (10 mg/night), or placebo. The primary outcome measure was the subjects' average intensity of jaw pain upon awakening over the prior week. This was recorded with a visual analog scale at pretreatment and at the completion of the 3-week trial. A secondary outcome measure was sleep quality based on the Pittsburg Sleep Quality Index. RESULTS: Within-group changes showed a statistically significant (P < .001) decrease in jaw pain upon awakening for all 3 groups. Between-group differences demonstrated a statistically significant difference (P < .016) between cyclobenzaprine and placebo, and between cyclobenzaprine and clonazepam. There was no significant effect on sleep quality in any group. CONCLUSION: This study suggests that cyclobenzaprine is statistically superior to either placebo or clonazepam when added to self-care and education for the management of jaw pain upon awakening. Based on the subjects' report of sleep quality, these medications failed to significantly improve sleep in the short term. PMID- 11889662 TI - Acupuncture and sham acupuncture reduce muscle pain in myofascial pain patients. AB - AIMS: To compare the effectiveness of dry needling in classically recognized acupuncture points ("acupuncture") with dry needling in skin areas not recognized as acupuncture points ("sham acupuncture") in reducing masseter muscle pain in a group of patients with myofascial pain of the jaw muscles. METHODS: Eighteen patients were randomly assigned to 1 of 2 experimental groups: Ten patients received acupuncture and 8 received sham acupuncture. A visual analog scale (VAS) was used to measure changes in masseter muscle pain evoked by mechanical stimulation of the masseter muscle before and after the experiment. RESULTS: Both groups showed a statistically significant reduction in VAS pain scores (P = .001). Seven out of 10 acupuncture subjects had a 10 mm or greater VAS reduction in pain, while 4 out of 8 of the sham acupuncture subjects had that great a pain reduction. There was no significant difference between the 2 groups. CONCLUSION: Both acupuncture and sham acupuncture reduced pain evoked by mechanical stimulation of the masseter muscles in myofascial pain patients. However, this reduction in pain was not dependent on whether the needling was performed in standard acupuncture points or in other areas of the skin. These results suggest that pain reduction resulting from a noxious stimulus (i.e., needling) may not be specific to the location of the stimulus as predicted by the classical acupuncture literature. PMID- 11889663 TI - Topical review: modulation of trigeminal sensory input in humans: mechanisms and clinical implications. AB - In this review, the modulatory effects of tooth and implant loading, orofacial pain, and psychological factors on somatosensory and jaw-motor function in humans are assessed. Experimental studies on the control of jaw actions have revealed that patients with prostheses supported by osseointegrated implants show an impairment of fine motor control of the mandible. One possibility is that this may be related to the loss of afferent information from periodontal ligament mechanoreceptors, which results in considerably higher and more variable forces to hold and manipulate food between the teeth. However, psychophysical investigations have shown that patients still perceive mechanical stimuli exerted on osseointegrated implants in the jawbone. The use of somatosensory evoked potentials may revealed what specific receptor groups are responsible for this so called osseoperception phenomenon. Orofacial pain is another modulator of trigeminal system functioning. Experimental jaw muscle pain has several effects on the somatosensory and motor function of the masticatory system, all of them serving to warn the individual about the ongoing damaging of tissues. Finally, the influence of mental state on the sensory and motor functions of the trigeminal system will be addressed. While some animal studies suggest that psychological stress can reduce acute pain, less speculative are the findings in human subjects that the anticipation of receiving a painful stimulus or undertaking difficult mental tasks can modulate jaw reflexes, including those evoked by mechanical stimuli applied to the teeth. Since such stimuli occur regularly during normal oral activities, the study of the resulting motor effects may yield clinically meaningful results in the context of other variables that modulate mandibular function. PMID- 11889664 TI - Dr. L.D. Pankey: the man and the legacy. PMID- 11889666 TI - Finding joy as a dental hygienist. PMID- 11889665 TI - Commitment to absolute values. PMID- 11889667 TI - A recollection and overview of the philosophy as taught by Dr. L.D. Pankey. PMID- 11889668 TI - One of Dr. Pankey's students who changed dentistry forever. PMID- 11889669 TI - Know yourself. PMID- 11889670 TI - Oral and Maxillofacial Pathology Case of the Month. Chronic hyperplastic candidiasis. PMID- 11889672 TI - Hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is a common malignancy worldwide and is a disease of multifactorial etiology. Strong correlations exist between the prevalence of the hepatitis B and C viruses and HCC incidence. HCC treatment may involve surgical resection, liver transplantation, locoregional treatments, and chemotherapy. Prevention of virus-related HCC is contingent upon control of hepatitis types B and C. Universal vaccination against hepatitis B could eliminate hepatitis B-related HCC; however, hepatitis C-related HCC still could occur because a vaccine for hepatitis C currently is not available. Individuals at risk for HCC should be screened for the disease. Early detection could result in improved prognosis and survival. PMID- 11889671 TI - A new class of antiemetic agents on the horizon. AB - This article reviews a new class of antiemetic agents, the neurokinin-1 (NK-1) receptor antagonists (RAs). Clinical trials of an NK-1 RA, aprepitant, are ongoing. PMID- 11889673 TI - OxyContin Use and Abuse. PMID- 11889674 TI - Intravenous Allopurinol. PMID- 11889675 TI - Glutamine: indicated in cancer care? AB - Glutamine is a nonessential amino acid contained in most dietary proteins and provides immune functions and fuel for the small intestine. For healthy people, dietary glutamine (from protein) usually is considered adequate. Results of research evaluating the potential benefits of glutamine during cancer therapy are encouraging but remain inconclusive. Some researchers have suggested recently that glutamine may, in fact, be a conditionally essential amino acid (Buchman, 2001). Decreases in glutamine levels after trauma or major burns, postoperatively, and in patients with diseases such as inflammatory bowel disease, AIDS, and cancer are widely recognized and acknowledged (Medina, 2001; Miller, 1999). The interpretation of data suggesting that glutamine supplementation is of benefit in almost any clinical situation is controversial. Additional research is needed to confirm the mechanism of action and efficacy of glutamine as adjuvant therapy in patients receiving cancer treatment. PMID- 11889676 TI - Large hyperpigmented perianal tumor. AB - Anorectal melanomas are rare tumors that occur more frequently with advancing age and peak in the sixth and seventh decades (Felz et al., 2001). Surgery is considered to be optimal therapy; however, chemotherapy and immunotherapy have been shown to be effective in a small number of patients. PMID- 11889677 TI - Imatinib mesylate. PMID- 11889678 TI - Families facing cancer: the forgotten priority. PMID- 11889679 TI - Hospital stay is a "teachable moment" for tobacco cessation. PMID- 11889680 TI - Taste alterations among patients with cancer. AB - Patients with cancer frequently develop taste alterations, which are manifested by food aversions and decreased caloric intake. Many etiologies are recognized, including the effect of tumors, cancer cell mitosis, vitamin deficiencies, and cytokine involvement. Preventing or improving taste alterations in patients with cancer is challenging. Clinicians play an important role in assessing, educating, and referring (when indicated) patients experiencing potential or actual taste alterations. Directions for further nursing research include the development of assessment tools and preventative strategies. PMID- 11889681 TI - Pathways for head and neck surgery: a patient-education tool. AB - Surgical treatment for head and neck cancer includes a variety of complex operative procedures. Patients scheduled for these operations often are presented with overwhelming amounts of information. Even after participating in the multidisciplinary, preoperative teaching process, they may be unable to fully comprehend the treatment plan. Pathways for head and neck surgery were designed to facilitate the patient- and family-education process and assist the patient in navigating through the surgical experience. This article discusses head and neck surgery pathway development, implementation, and outcome evaluation. PMID- 11889682 TI - Risk factors and health promotion in families of patients with breast cancer. AB - Women with a family history of breast cancer have an increased risk of developing the disease. Women identified as "high risk" for developing breast cancer have been shown to exhibit increased levels of psychological distress and anxiety related to breast cancer. Oncology nurses can address this barrier and others, such as altered risk perception and lack of physician recommendation for screening. Oncology nurses also can identify high-risk families that may be candidates for genetic testing for breast cancer susceptibility, provide comprehensive teaching about breast self-examination (BSE), and clarify misconceptions about early detection. Primary prevention measures for hereditary breast cancer include prophylactic mastectomy and oophorectomy and chemopreventative agents. Secondary prevention measures include screening and early detection with mammography, clinical breast examinations, and BSE. Nurses have a responsibility to educate families of patients with breast cancer about risk factors, primary and secondary preventive measures, genetic testing, and screening recommendations. PMID- 11889683 TI - Osteoporosis in men treated with androgen suppression therapy for prostate cancer. AB - Men with advanced or metastatic prostate cancer commonly receive long-term treatment with luteinizing hormone-releasing hormone (LHRH) agonist therapy. This prolonged treatment causes a hypogonadal state of chronic testosterone deficiency. Similar to estrogen deficiency in postmenopausal women, testosterone deficiency among these men negatively affects bone metabolism through a complex self-regulating, negative feedback system and subsequent reduction in bone formation. If left undetected or untreated, the risk for osteoporosis rises. Osteoporosis increases the likelihood of fracture, especially of the hips. Researchers are studying the effects of LHRH agonist therapy on osteoporosis and other related conditions to determine whether interventions, such as pharmacologic agents (e.g., bisphosphonates), dietary supplements (e.g., calcium, vitamin D), and exercise, can slow or prevent the process and assist healthcare providers in knowing how to counsel patients. Current recommendations are found in the literature on glucocorticoid-induced and menopausal osteoporosis. Nurses need to stay abreast of current knowledge in this area, as it is expanding rapidly. PMID- 11889685 TI - HIV/AIDS update. PMID- 11889686 TI - Primary care: promoting sexual health and HIV prevention through risk assessment and client education. AB - After 20 years, human immunodeficiency virus (HIV) is still a global health problem, mandating the need to incorporate HIV prevention into primary care. This report presents tips and examples to assist in conducting a clinical, drug use, and sexual risk assessment, using the term TOSSIS to cover HIV transmission sources. Nurse practitioners can also assist in HIV prevention by serving as patient and parent educators. PMID- 11889684 TI - Chemotherapy-induced nausea and vomiting. AB - Nausea and vomiting (N&V) is among the most distressing side effects of chemotherapy, despite the development of more efficacious antiemetic agents. As many as 60% of patients who receive cancer chemotherapy experience some degree of N&V. However, the actual incidence is difficult to determine with accuracy because of the variety of drugs, doses, and health conditions of the patients who receive cancer treatments. This article examines the state of the science related to chemotherapy-induced nausea and vomiting and reviews both pharmacologic and behavioral strategies that have demonstrated efficacy in managing these distressing symptoms. PMID- 11889687 TI - HIV postexposure prophylaxis after sexual assault. AB - Recent advances in the treatment of human immunodeficiency virus (HIV) disease have prompted health care providers to reexamine recommendations for prophylaxis of HIV infection. Parallels with occupational exposure through mucous membrane tissues spur consideration of HIV prophylaxis after sexual assault for several reasons. In both instances, exposure occurs at a single point in time and is unlikely to recur. Although the Centers for Disease Control and Prevention does not make definitive recommendations regarding postexposure prophylaxis after sexual assault, the reality is that as clinicians, we face situations in which we must consider treatment for prevention of HIV disease after sexual assault. Guidelines for treatment and how to create and implement a policy to ensure the best outcomes, and provide a high quality of patient care with the New York State guidelines as a model, are discussed. PMID- 11889688 TI - Complications of HIV: lipodystrophy, anemia, renal, cardiovascular, and bone diseases. AB - Diagnosis of a human immunodeficiency virus (HIV) infection has long been associated with a short life expectancy, with care centered on the treatment and prevention of opportunistic infections and symptom management. As progress has been made through better understanding of the virus, and improved medications and treatments, persons with HIV are living longer, more productive lives. In addition to the scientific breakthroughs in treating the HIV-infected patient, more is also being learned about the long-term effects of the treatments and the virus, such as lipodystrophy syndrome. Comorbid diseases are also becoming more common in patients with HIV, such as anemia, bone disease, renal disease, and cardiac disease, which may be the result of metabolic complications or HIV itself. There is a vast array of comorbid diseases and adverse effects of medications in persons with HIV disease; and information regarding these issues is constantly changing as research continues to show more about their pathophysiology and treatment options. The most common comorbid disorders will be explored. A detailed explanation of the lipodystrophy syndrome and its common metabolic manifestations will be presented, including current research. In addition, the etiology, clinical findings, diagnosis, and treatment of the most common comorbid diseases associated with anemia and the bone, renal, and cardiovascular systems will be presented as they relate to the HIV-infected patient. All of the descriptions are designed for the primary care provider, who may be in contact with an HIV-infected patient whose differential diagnoses consist of one or more of these disorders. PMID- 11889689 TI - Primary care of HPV management in HIV-infected women. AB - The natural history of human papillomavirus (HPV) differs in women infected with (human immunodeficiency virus) HIV when compared with the general population. This report provides insight into the complexities of treating HPV infection and the differences found in HIV-infected women. By understanding the challenges associated with this opportunistic infection in HIV-infected women, nurse practitioners will be better prepared to provide primary care to this specific population. PMID- 11889690 TI - Promoting healthy behaviors in HIV primary care. AB - Recent treatment advances have prolonged the life expectancy of persons with human immunodeficiency virus (HIV). HIV care providers must now promote healthy behaviors, such as smoking cessation, exercise, and screening for general medical problems, such as diabetes and hyperlipidemia. This report describes recently published evidence and recommendations for providing HIV primary care. PMID- 11889692 TI - [Morphology of vascularized tissue complexes after their transplantation on the vascular pedicle]. AB - Light microscopy was used to study microvascular bed and tissue leukocytes in transplanted tissue complex on the vascular pedicle. The cause of the transplant edema was disturbed lymph flow manifesting in a significant dilation of lymph vessels and interstitial spaces. Restoration of the lymph flow and hemocirculation takes place by formation of new vessels registered in all the transplant tissue 2-8 months after transplantation. Late after the transplantation the tissues of the graft underwent progressive sclerotic transformation due to denervation and loss of specific functions of the transplanted tissues. PMID- 11889691 TI - [The role of EGF-stimulated epidermis in the regulation of wound healing]. AB - Sluggish wounds are characterized by impaired proportions of proinflammatory cytokines, deficiency of fibrogenic growth factors, imbalance in the system of matrix metalloproteinases and their inhibitors this preventing reparation. The study was made of biopsies obtained from patients with sluggish wounds before the treatment, 5, 10 and 15 days after transplantation on the wound of allogenic EGF stimulated cryopreserved epidermis. The wound closure with biologically active coat was followed by the reduction of expression of proinflammatory cytokines and return to their normal correlations, higher production of fibrogenic growth factors, restoration of balance in the expression of MMP-9 and TIMP-1/TIMP-2. PMID- 11889693 TI - [Changes in the components of the extracellular matrix and its regulators in the endometrium of women with habitual abortion]. AB - Biopsies of the endometrium from 93 patients with habitual abortions (HA) were studied immunohistochemically. Chronic endometritis (CE) was diagnosed in 26 patients. Decreased tenascin expression in the endometrial stroma with merozin accumulation in the glandular basal membranes, covering epithelium and vessels were found in HA combined with CE. Abnormal distribution of collagen type I and III with predominance of type I collagen and appearance of merozin. This is indicative of alteration in the endometrium maturation which may result in inadequate ovum implantation and pregnancy interruption. Stromal alterations in HA with or without chronic endometritis are under multifactorial control of various mediators. High level of MMP-9 combined with low level of TFP-beta 1 may serve a prognostic criterion of successful completion of pregnancy regardless of CE. Low level of MMP-9 may increase the risk of pregnancy interruption. PMID- 11889694 TI - [Expression of the protein marker p16INK4a in the cervix uteri cancer]. AB - Immunohistochemical study was carried out of 18 cervical carcinomas (13 squamous and 5 adenomatous) and of 3 cases of cervical intraepithelial dysplasia. Formalin fixed paraffin-embedded tissue samples from biopsies as well as from surgical material were used. Staining was performed with monoclonal antibodies to protein p16INK4a. Cytologic smears of epithelial cervical cells from 7 healthy women were taken as a negative control. The reference group consisted of 5 cancer patients with other tumors (breast cancer, B-cell lymphoma). Overexpression of p16INK4a was registered in cervical cancer. PMID- 11889695 TI - [Morphological and immunohistochemical characteristics of primary brain lymphomas in AIDS patients]. AB - Primary lymphomas of the brain were studied in 6 AIDS patients. The lymphomas were characterized by multicentricity, extended necroses and location in large hemispheres. Two types of tumor cell growth were recorded: diffuse and perivascular with vascular wall destruction. According to the International Histologic Classification (2001), these lymphomas were classified as diffuse B cell large cell lymphomas subdivided into 3 variants: immunoblastic with plasma cell differentiation, centroblastic polymorphic and large cell anaplastic (by Kils classification criteria). Immunohistochemically (LMP-1) and by hybridization in situ (RNA EBER-1,2) Epstein-Barr virus was found in tumor cells from all the patients. In 5 of 6 cases studied expression of the antiapoptotic protein bcl-2 and in 2 cases expression of mutant p53 were revealed. PMID- 11889696 TI - [Proliferative markers of meningiomas: immunohistochemical analysis and prognostic value]. AB - Routine pathological examination cannot precisely predict the clinical course of meningiomas because even histologically benign tumors may recur after total removal. And so, numerous efforts have been made for evaluation of meningioma growth fraction and its prognostic value. In this study the prognostic significance of DNA toposiomerase II alpha (topoII) and cyclin A immunohistochemistry was examined in a series of 263 meningiomas. The topoII and cyclin A scores exhibited a close correlation with Ki-67 immunostaining. Significant differences between the indices for all the three markers were noted among the three grades of meningiomas. The scores for all the three markers were significantly different between recurrent and non-recurrent meningiomas including removed benign tumors. The most important information for an individual clinical outcome of histologically benign meningiomas should be elicited from simultaneous evaluation of all the three proliferative markers. PMID- 11889697 TI - [Morphological changes of internal organs in opium addiction]. AB - Morphological manifestations of chronic opium addiction are presented. Specific features of pathological processes in drug abusers may facilitate forensic medical and postmortem diagnosis of narcotic drug poisoning or other death causes in chronic narcotic intoxication. PMID- 11889699 TI - [Morphological diagnosis of various helminthiases during analysis of surgical specimens of appendices]. AB - Appendices with intact content after appendectomy were studied histologically. On the appendixes content sections fragments of helminths parasitizing in the colon as well as their ova were often found. Determination of helminth invasion nature raises the efficiency of the pathologist's examination. PMID- 11889698 TI - [Correlation between proliferative processes and cell death in non-small cell lung cancer with glandular differentiation at different stages of tumor progression]. AB - Surgical material (removed lungs or their parts) from 58 patients operated in 1993-1998 was investigated. Lung adenocarcinomas (LAC) are characterized by low proliferative activity of tumor cells. With a decrease of LAC differentiation, tumor cell death by terminal differentiation also diminished which was accompanied by low bcl-2 expression and enhancement of spontaneous apoptosis with active accumulation of protein products of p53 expression in tumor cells nuclei. Expression of c-myc and bax remained unchanged. On the whole, the picture reminds that in lung squamous cell carcinoma. Bronchioloalveolar carcinoma is characterized by low proliferative activity combined with higher apoptosis compared to LAC. Large cell lung carcinoma and adenomatous-squamous lung carcinoma demonstrated the highest proliferation and spontaneous apoptosis of tumor cells with accumulation in these cells of p53, bcl-2 and bax comparing to non-small cell lung carcinoma (NSLC) with adenomatous differentiation. Progression of NCLC with adenomatous differentiation largely depends not only on proliferative activity of tumor cells but on tumor cell death due to terminal differentiation, apoptosis and necrosis as well. PMID- 11889700 TI - [Malignant carcinoid of the urinary bladder]. AB - The paper reports a case of a rare tumor of diffuse endocrine system--a malignant carcinoid of the urinary bladder. PMID- 11889701 TI - [Rupture of thoracic aorta aneurysm 6 years after endoprosthetic surgery]. AB - A case of 6-year stay of a nitinol stent in the lumen of atherosclerotic aneurysm of the descending thoracic aorta is reported. Problems of the prosthesis integration into the organism, nitinol are activity and toughness as well as causes of the aneurysm rupture are discussed. Literature data on endovascular treatment of the aortic aneurysm including in the beginning of its rupture and in old persons with grave concomitant diseases are presented. PMID- 11889702 TI - [Spitz nevus (juvenile nevus) of the penile skin]. AB - Compound spindle cell and epithelioid cell nevus (Spitz nevus) at the anterior surface of the penile skin in a 31-year-old patient is described. The histological structure of the tumor is discussed in terms of differential diagnosis. PMID- 11889704 TI - [Hibernation and stunning as manifestations of ischemic dysfunction of the myocardium]. AB - Current information on hybernation and stunning which are the basis of myocardial dysfunction resulting from chronic ischemic heart disease and revascularization is provided. Myocardial stunning is an acute disturbance of a contractile function of ischemic myocardium at the moment of coronary circulation restoration done by various methods (coronary shunting, angioplasty, thrombolysis). Myocardial hybernation is a chronically developing foci of subnormal contractility in the region of stenotic artery. The difference between them is that stunning is a complex of structural and metabolic damages under the condition "ischemia-reperfusion" while hybernation is an adaptation of the myocardium to chronic ischemia by metabolism switching to anaerobic glycolysis. The study of pathophysiology and morphology of hybernating and stunned myocardium is necessary for developing methods of myocardium protection from ischemic damage. PMID- 11889705 TI - [Vascular pathology in antiphospholipid syndrome]. AB - Antiphospholipid syndrome (APLS) is characterized by the presence of antibodies against phospholipids in circulating blood, recurrent arterial and/or venous thrombosis, recurrent and spontaneous abortions and thrombocytopenia. This syndrome develops in the presence of autoimmune diseases, most frequently lupus erythematosus, neoplastic and infectious processes. Circulating antibodies damage endothelium of the vessels and result in active thrombogenesis and slow fibrinolysis. Morphologically, APLS presents with thrombosis, focal angiomatosis, hemorrhage, endothelium and vascular wall alterations in the absence of inflammatory cell reaction. These lesions may be associated with vasculitis in various organs. PMID- 11889706 TI - [Effect of opioid peptides on the repair regeneration of the bone tissue]. AB - The study is made of the effects of selective agonists of opioid receptors (DAGO- the agonist of mu-receptors, DSLET--the agonist of delta-receptors, dynorphin A,I 13,--the agonist of kappa-receptors) on reparative osteogenesis in CBA mice. The drugs were injected intraperitoneally within 7 days after bone fracture in equimolar doses: DAGO--6.3 mcg/kg, DSLET--10 mcg/kg, dynorphin A (1-13)--20.2 mcg/kg. It was established that the use of DSLET and DAGO accelerates the development of newly-synthesized spongy bone tissue in the bone regenerate. This manifested with its early appearance (4 days after the fracture) as well as higher indices of specific volumes of bone tissue in the regenerate within the experiment (7, 10, 14 days after bone fracture) in comparison with the control animal group. Dynorphin A(1-13) had no influence on regenerate formation in our experiments. PMID- 11889708 TI - Statistical contributions to molecular biology. PMID- 11889707 TI - [Morphological and immunohistochemical characteristics of repair processes in nonhealing wounds]. AB - Sluggish wounds (SW) are characterized by chronic inflammation, abnormal proportion of extracellular matrix (ECM) components, appearance of ECM components that do not occur under normal conditions, this preventing formation of basal membrane (BM) and reepithelization of the wound. 89 biopsies from 25 patients with SW were studied before treatment and in different intervals after transplantation of allogenic cryopreserved epithelium on the wound. Decreased collagen type III content and accumulation of tenastcin in combination with low number of myofibroblasts were observed in ECM before the transplantation. Normalization of quantitative and qualitative composition of ECM components, BM formation, wound reepithelization without scarring were noted after the transplantation; myofibroblast number remained very low. Wound healing without contraction and scarring is a distinctive feature of reparative process in SW when allogenic cryopreserved epidermis is used. PMID- 11889709 TI - Power and sample size considerations in molecular biology. PMID- 11889710 TI - Models for determining genetic susceptibility and predicting outcome. PMID- 11889711 TI - Multiple tests for genetic effects in association studies. PMID- 11889712 TI - Linking image quantitation and data analysis. PMID- 11889713 TI - Statistical considerations in assessing molecular markers for cancer prognosis and treatment efficacy. PMID- 11889714 TI - Power of the rank test for multi-strata case-control studies with ordinal exposure variables. PMID- 11889715 TI - Introduction to microarray experimentation and analysis. PMID- 11889716 TI - Statistical methods for proteomics. PMID- 11889717 TI - Statistical methods for assessing biomarkers. PMID- 11889718 TI - Recognizing the needs of the underserved in light of public health projects. AB - Healthy People 2000 grew out of the idea that positive changes could occur if Americans worked together toward certain goals. One of the most compelling lessons learned from the Healthy People 2000 initiative is that as a nation we can make dramatic progress in a relatively short period of time. Ten years later, Healthy People 2010 evolved from the underlying belief that the health of the individual is inseparable from the health of the community, and that the health of every community determines the overall health status of the nation. Healthy People 2010 challenges individuals, communities, and healthcare professionals to take specific steps to ensure that good health, as well as long life, is accessible to all Americans. Critics of public health projects such as Healthy People 2010 argue that these projects often fail to meet their goals because solutions are seen through the eyes of the dominant culture--that we have a tendency to disregard the lived experience of those we are trying to reach. This is the reason why a deeper cultural awareness is essential to public policy formation. The real challenge and opportunity for wound care clinicians are to recognize the thousands of alternative health and healing systems in the United States, each with a colorful and unique worldview. This article identifies public health projects that impact clinical care, defines what is meant by the term underserved, and describes the health practices of what are considered ethnically diverse groups. PMID- 11889719 TI - Heparin-induced thrombocytopenia as the cause of gluteus muscle necrosis: a case study describing the benefits of multidisciplinary physical and psychosocial interventions. AB - Heparin-induced thrombocytopenia, an increasingly recognized aspect of heparin therapy, occurs in 0.6% to 30% of patients receiving heparin. Approximately 1% of those patients develop the more severe heparin-induced thrombocytopenia II, also called white clot syndrome, in which synchronous venous and arterial thrombi impede blood flow in central vessels. Mortality (as high as 25%) and morbidity are related to the site and extent of thrombi formation. Following vascular surgery, one patient manifested an unusual consequence of heparin-induced thrombocytopenia II when thrombotic blockage of the vessels supplying the bilateral gluteus maximus and minimus muscles resulted in tissue ischemia and death. Supporting the patient through numerous complications and managing the extensive wound healing process required multidisciplinary skills and innovative technology, including use of vacuum-assisted closure therapy, platelet-derived growth factors, parenteral and enteral nutritional support, and spirit-restoring favorite foods. This article describes the patient's life-threatening and long lasting effects from an allergic reaction to heparin therapy. In addition to information about the diagnosis of heparin-induced thrombocytopenia, this article also describes management of the wound and other aspects of care and comforting that occurred over a 9-month hospitalization. PMID- 11889720 TI - The role of phytotherapeutics in wound management. AB - This paper outlines the history and development of herbalism. The fall of medicinal herbs from favor and how recently they have returned to prominence are discussed, as well as the role of phytotherapy in successfully resolving chronic disease processes and managing wounds. The role of the herbalist is explained and includes a detailed description of the techniques employed by practitioners. Seven basic herbs are profiled regarding benefits, therapeutic potential, interactions, and effects. A hydrotherapy procedure for wound management is included. PMID- 11889721 TI - Assessment of venous leg ulcers: an in-depth discussion of a literature-guided approach. AB - Venous leg ulcers represent a significant public health problem that will increase as the population ages. The elderly, the most likely to be afflicted by this condition, present the clinician with special challenges. The diagnosis of venous leg ulcers involves taking a careful history, paying attention to the existence of factors that predispose individuals to the development of chronic venous insufficiency. Clinical features of venous insufficiency are important because their recognition allows clinicians to distinguish venous from other chronic ulcers. An essential part of the assessment of all patients with chronic wounds is an evaluation of intercurrent diseases, common in the elderly, which may impact on the wound healing process. Thus, in addition to managing venous insufficiency and the wound bed, all other factors, systemic and local, that may impede healing need to be investigated and corrected if necessary. Social and psychological issues common to all chronic illnesses need to be addressed as well. This holistic approach should be standard practice and is applicable to the assessment and management of all chronic leg ulcers. This often requires coordinating a multidisciplinary team of wound healing caregivers. More work needs to be done to clarify a few issues because areas of controversy persist. Although a great deal is known about the effects of vitamin deficiency on acute wounds, less is known about chronic ulcers. Similarly, the role of vitamin supplementation in managing chronic ulcers needs further study. Guidelines are needed to determine indications for wound culturing. In addition, more study is required to establish the most effective means of obtaining quantitative cultures. However, the relationship between bacteria and chronic wound healing goes beyond simple quantitation and other factors such as bacterial virulence and host resistance. These controversial issues will be reviewed. Treatment and prevention of venous leg ulcers will be discussed in a subsequent article. PMID- 11889723 TI - Rethinking our approach to pressure ulcers. PMID- 11889722 TI - Assessing and treating pelvic organ prolapse. AB - At least half of all women who have given birth experience pelvic organ prolapse, a condition where pelvic organs protrude through the vagina. Because of the presentation of the different aspects of prolapse, treatment had become compartmentalized in line with pelvic involvement, with urologists, gynecologists, colorectal surgeons, and gastroenterologists each addressing their field of expertise. In addition, urinary or fecal incontinence, urinary retention, and urinary tract infections often are associated with pelvic organ prolapse. Both pelvic organ prolapse and incontinence have a significant impact on the quality of life. New training programs in urogynecology and reconstructive pelvic surgery are producing clinicians who are better equipped to treat pelvic organ prolapse, as well as related urinary and fecal incontinence. This article provides an overview of the various aspects of pelvic organ prolapse for all clinicians involved in assessment, treatment, and potential prevention of this condition. PMID- 11889724 TI - What's wrong with our healthcare system. PMID- 11889725 TI - Using epidemiology in patient education for post-prostatectomy urinary incontinence. AB - This article reviews and discusses the prevalence and incidence of and risk factors for urinary incontinence after prostate cancer surgery. The reported prevalence rates of urinary incontinence in men vary among studies due to different definitions and methodologies; however, it is agreed that urinary incontinence is a common condition, especially immediately after surgery. Although few risk factors have been identified, damage to the sphincter or its nerves is an accepted underlying pathology. Urinary incontinence can be devastating to men and sensitivity to emotional and educational needs is critical. PMID- 11889726 TI - Behavioral treatments for post-prostatectomy incontinence. AB - Despite improvements in surgical approaches to radical prostatectomy, many patients experience moderate to severe urinary incontinence during the first few postoperative weeks. For some patients, leakage continues for several months or years. Urinary incontinence has a significant impact on quality of life in these typically active patients. Surgical interventions are not recommended until after a 6 to 12 month trial of behavioral interventions. Behavioral interventions include supportive care, diet and medication management, pelvic floor rehabilitation, inhibition techniques, and patient education and support. This integrative review of the literature summarizes current knowledge of behavioral interventions for post-prostatectomy urinary incontinence for the primary care and home care clinician. Aspects of care specific to masculine culture are explored. Recommendations for further research include determination of optimal timing and protocols for pelvic floor rehabilitation, best practices for patient and family education, and development of a standard definition of urinary incontinence so the efficacy of treatments can be compared. PMID- 11889728 TI - Urinary health in eldercare environments: an update from the NAFC. AB - The National Association For Continence elected to cancel its 2001 conference scheduled for October 2001 in Washington, DC in light of national events. Executive Director Nancy Muller has provided Ostomy/Wound Management with a synopsis of key issues, important considerations, and the syllabus. We are grateful for the opportunity to serve as a forum for this worthy organization. PMID- 11889727 TI - Urinary incontinence knowledge and attitudes in Sao Paulo. AB - Urinary incontinence is a social and hygienic problem that affects 15% to 35% of individuals aged 60 years or older, with an estimated annual cost in the United States of $16 billion. To better understand the effects of misinformation and underestimation of the problem, a study to identify knowledge and attitudes related to urinary incontinence was conducted among 400 community-dwelling residents (55 years of age or older). The instrument covered four knowledge/attitude subject areas: treatment and effects, causes, the relationship between aging and urinary incontinence, and physician/patient discussion regarding urinary incontinence. Interviews were conducted in waiting rooms, shopping centers, subways, and hospital/clinic waiting rooms. One hundred, forty two people (43% women and 23% men) reported current or previous bladder problems. Only two of the 14 knowledge statements elicited a correct response from the majority (60%) of participants. The percentage of correct answers to the other 12 statements ranged from 23% to 43%. Most people believed that urinary incontinence is an inevitable consequence of aging, and the majority of treatment responses (58%) indicated a lack of knowledge about available treatments. Differences in the percentage of correct responses related to the treatment and causes of urinary incontinence were observed between respondents with varying educational levels (P = 0.001) and between Caucasian and African American respondents (P = 0.001). The results confirm that misinformation about urinary incontinence is common in the general population and that educational interventions are needed if the prevalence of uncontrolled urinary incontinence is to be solved. PMID- 11889729 TI - Home health PPS is here. PMID- 11889730 TI - Politics, public policy, and healthcare. AB - In many cases, justice is the ethical principle that drives decisions around politics, public policy, and healthcare. This article presents the ethical principle of justice as it relates to the equitable distribution of healthcare goods and services. The impact of politics, especially in a presidential election year, is discussed as it relates to agendas and alternatives for healthcare policy formation--specifically, as it directly and indirectly influences wound care and wound care delivery. Opportunities for wound care clinicians are presented. PMID- 11889731 TI - A comparative study of the effects of UVC irradiation on select procaryotic and eucaryotic wound pathogens. AB - Managing wounds infected with a mixture of several types of microorganisms such as bacteria (procaryotes) and fungi (eucaryotes) is a challenging clinical situation. The purpose of this study was to determine the effectiveness of ultraviolet light (UVC) in eradicating select procaryotic and eucaryotic organisms, in both pure culture and mixed cultures in vitro. Five replications of each organism or mixture of organisms (10(6) organisms/mL singly or 10(15) organisms/mL mixed culture) were plated. The cultures were treated with a UVC light 1 inch from the surface. Irradiation times were 0, 2, 3, 4, 5, 15, 30, 45, 60, 90, 120, and 180 seconds. Bacterial cultures were incubated and colony counts performed. Upon exposure to UVC, a 99.9% kill rate was obtained at 3 to 5 seconds for the procaryotic organisms (Pseudomonas aeruginosa and Mycobacterium abscessus) tested. However, 15 to 30 seconds of UVC treatment was required to obtain 99.9% kill of the eucaryotic organisms (Candida albicans, Aspergillus fumigatus) tested. This study demonstrates a decreasing sensitivity of evolutionarily more complex organisms to UVC. This study also provides further evidence that short exposure times to UVC are detrimental to procaryotic and simple unicellular eucaryotic organisms while sparing more complex multicellular organisms. PMID- 11889732 TI - Calciphylaxis and its relation to end-stage renal disease: a literature review and case presentation. AB - Calciphylaxis is an uncommon phenomenon usually observed in patients with chronic renal failure who are on dialysis and who have secondary hyperparathyroidism. Treatment is multifocal and may include wound debridement, wound excision, and various methods of achieving closure. Removal of the parathyroid glands often is advocated, but this option remains controversial. Despite treatment, there is a fatal outcome in a majority of cases due to septicemia. Prompt recognition and treatment are imperative to help prevent septicemia. A review of the literature and two case studies are presented. PMID- 11889733 TI - The impact of telemedicine on outcomes of chronic wounds in the home care setting. AB - Since 1997, more than 2,700 home care agencies have closed due to changes in reimbursement. With the implementation of a prospective payment system, there is concern--not only regarding the survival of the remaining home care agencies, but also whether adequate care can be provided to the patient. Chronic wound care is both prevalent and costly in home care. Factors contributing to cost include inconsistency of wound assessment and documentation and low usage of advanced wound products. These factors lead to lengthened healing time, more frequent visits by practitioners, and low healing rates. Involving a wound specialist can improve patient outcomes while decreasing cost. The authors examined the utilization of telemedicine in situations where wound specialists consulted with the home health nurse in the patient's home regarding care of chronic wounds. During the two-way video visit, the wound specialist assessed the patient and the wounds and made recommendations for treatment. The wound specialist also collected outcome data during the visits. This data was then compared with like data collected as a baseline prior to the telemedicine intervention. Results revealed improved healing rates, decreased healing time, decreased number of home health visits, and a decreased number of hospitalizations related to wound complications. Telemedicine was deemed a viable option for delivering quality, cost-effective care to chronic wound patients in the home care setting. PMID- 11889734 TI - To provide good care, you have to get in the game. PMID- 11889735 TI - Preparing the wound bed--debridement, bacterial balance, and moisture balance. AB - Successful diagnosis and treatment of patients with chronic wounds involve holistic care and a team approach. The integration of the work of an interdisciplinary care team that includes doctors, nurses, and allied health professionals with the patient, family, significant others, and caregivers offers an optimal formula for achieving wound resolution. Such an approach challenges practitioners and everyone participating in wound care to integrate data and information that arise from a number of sources and mitigating factors. In this article, the authors define the changing paradigm that links treatment of the cause and focuses on three components of local wound care: debridement, wound friendly moist interactive dressings, and bacterial balance. The authors demonstrate that the treatment of chronic wounds can be accomplished through a series of recommendations and rationales based on the literature and their experience. These recommendations lay the groundwork for thorough assessment and evaluation of the wound. PMID- 11889736 TI - Best practices for the prevention and treatment of pressure ulcers. AB - In this article, the Canadian Association of Wound Care puts forward 12 recommendations for best practices in the prevention and treatment of pressure ulcers that focus on an interdisciplinary patient-centered approach. These recommendations are a synthesis of the Agency for Health Care Policy and Research guidelines, European guidelines, and current literature as interpreted by the Canadian experience and achieved through a national consensus panel. The article concludes that best practice guidelines must be fluid documents that respond to new evidence and experience. PMID- 11889737 TI - Best practices for the prevention, diagnosis, and treatment of diabetic foot ulcers. AB - Although the prevention, assessment, and treatment of diabetic foot ulcers has improved in recent years, care is often fragmented and does not always meet best clinical practice. This article incorporates current best clinical practices and expert opinion with available research to arrive at 11 recommendations. These recommendations include adequate vascular supply, infection control, pressure offloading, and an optimal local wound environment. This approach is best accomplished through a multidisciplinary team and revolves around the active participation of the person with diabetes. The authors' intent is to provide a practical, easy-to-follow guide to allow healthcare professionals to establish and empower a team to provide best clinical practices. PMID- 11889738 TI - Expert opinion to link the scientific literature to patient care. PMID- 11889739 TI - The role and source of power. PMID- 11889740 TI - Artificial intelligence: the clinician of the future. AB - Human beings have long been fascinated with the idea of artificial intelligence. This fascination is fueled by popular films such as Stanley Kubrick's 2001: A Space Odyssey and Stephen Spielberg's recent film, AI. However intriguing artificial intelligence may be, Hubert and Spencer Dreyfus contend that qualities exist that are uniquely human--the qualities thought to be inaccessible to the computer "mind." Patricia Benner further investigated the qualities that guide clinicians in making decisions and assessments that are not entirely evidence based or grounded in scientific data. Perhaps it is the intuitive nature of the human being that separates us from the machine. The state of artificial intelligence is described herein, along with a discussion of computerized clinical decision-making and the role of the human being in these decisions. PMID- 11889741 TI - Nutrition and its role in wound healing. PMID- 11889742 TI - Utilizing a systems approach to implement pressure ulcer prediction and prevention. AB - Efficient, effective patient care is an objective shared by all healthcare settings and systems. It is generally accepted that using national clinical guidelines facilitates pursuit of this objective. However, implementation of a guideline, or any process improvement activity, requires a systematic, collaborative approach from which new processes can be purposefully designed. This article reviews systems theory, presents the steps for process improvement using the Plan/Do/Check/Act cycle, and references a recent statewide quality improvement study conducted by the authors in collaboration with Stratis Health, a Minnesota Medical Peer Review Organization. PMID- 11889743 TI - "Constant force technology" versus low-air-loss therapy in the treatment of pressure ulcers. AB - "Least costly but most effective" has never been more important to healthcare delivery than in the current healthcare environment of changing reimbursement systems. In wound care, the high costs associated with renting advanced support surfaces may be an area where expenses can be decreased if similar outcomes can be attained by using more affordable mattresses. This article compares the healing rates and eventual outcomes of wounds among two groups of patients randomly assigned to one of two support surfaces. A commonly used low-air-loss mattress was compared with the study mattress, an advanced, non-powered, air and foam surface. Subjects were patients admitted to either of two long-term healthcare settings for the treatment of wounds. The resulting two groups of 10 patients each were evenly matched for average age and percentage of patients who were nutritionally deficient as indicated by albumin or pre-albumin levels. The presence of gastrointestinal tubes and ventilator dependency also was recorded. Consistent wound care protocols were used on both groups, including turning schedules, nutrition, topical medication, and dressings. The study period covered a maximum of 8 weeks; interest was centered on the rate of wound healing and progress toward a goal rather than the length of time to completely close a wound. After 8 weeks, or upon discharge from the study pressure ulcers in the study mattress group closed at an average rate per week of 9.0% +/- 4.8 versus 5.0% +/- 3.7 in the low-air-loss mattress group. This study indicates that the study mattress can provide benefits to the wound healing process similar to or better than low-air-loss mattresses at a substantially reduced cost. PMID- 11889744 TI - Contact dermatitis treated with new topical products: a case study. AB - Contact dermatitis, an inflammatory response of the skin to an irritant or an allergen, can affect hospital staff. Most clinicians are routinely exposed to irritants such as latex, detergents, and chemicals. Treatment with topical corticosteroids and avoidance of suspect irritants usually resolves the dermatitis. A case study is presented of a licensed practical nurse who developed persistent contact dermatitis. The dermatitis did not resolve with 15 months of traditional treatments. Only after 3 months of treatment with two investigational topical products, which are now available to the public, was the dermatitis resolved and complete healing achieved. This case study discusses the new products and traditional treatment products used and presents results of irritant specificity testing and a series of photographs documenting resolution and healing. PMID- 11889745 TI - Healing: can we? Must we? Should we? PMID- 11889746 TI - Home health FAQs: (finding abundant quirks). PMID- 11889747 TI - Walking the HMO balance. AB - Fidelity is the ethical obligation to act in good faith to keep promises, fulfill agreements, and maintain relationships and fiduciary responsibilities. Consumers are increasingly interested in the balance between the fiscal viability of our current healthcare delivery system and the system's reason for existence--that is, to serve the health needs of clients. Escalating healthcare costs have driven many institutions and third party payors to examine service and payment practices. Some consumers and consumer rights groups contend that these evolving practices threaten the very essence of health and healthcare. The ethical obligation of fidelity, especially as it relates to the business model of healthcare, is examined. Threats to fidelity are reviewed, and the response to these threats by one consumer rights group is presented. A case study is included. PMID- 11889748 TI - Osteomyelitis in the diabetic patient: diagnosis and treatment. Part 2: Medical, surgical, and alternative treatments. AB - In the diabetic population, wound and foot infections are often mixed, containing from three to six organisms. This creates a significant problem regarding antibiotic protocols. Many of these episodes result in contiguous bone infections with subsequent erosive changes, sequestra, and involucrum. A multidisciplinary approach to treatment is often required. Studies have shown that a protocol of 6 weeks of intravenous antibiotics may be appropriate to treat osteomyelitis; however, this is controversial and often not curative. Osteomyelitis is a surgical disease; a subtle balance between medical and surgical therapy is necessary if a potentially curative outcome is to be achieved. The duration of antibiotic therapy may be shortened considerably after surgical intervention. In cases of infection mitigated by severe peripheral vascular disease, end-stage renal disease, diabetes, or other medical problems where surgery is not an option, long-term antibiotics may be used as suppressive therapy along with adjunctive local treatments. The following is the second article of a two-part series. The first paper discussed the diagnosis and microbiology of contiguous osteomyelitis in the diabetic foot. This article outlines the various medical, antibiotic, and surgical options available to the clinician. Adjunctive and alternative therapies also are discussed. PMID- 11889749 TI - Impact of practice guidelines on support surface selection, incidence of pressure ulcers, and fiscal dollars. AB - Predicated on a need to control overall hospital costs and to integrate a Level 1 trauma center (Campus A) with a family practice based tertiary care hospital system (Campus B), expenditures associated with rental support surfaces were evaluated. Consistency and appropriateness of support surface selection is necessary to promote positive clinical outcomes, patient comfort, and a healthier bottom line, despite increasing costs. Clinical practice guidelines for therapeutic support surfaces were developed to decrease support surface expenditures and maintain prevalence rates below national averages. Utilizing the Agency for Health Care Policy and Research algorithm for managing tissue loads, along with other guidelines, criteria for prevention, comfort, and treatment were developed to assist nurses and physicians in support surface selections. A prevalence study was conducted before these criteria were implemented and repeated 1 year later. Expenditures for all rental support surfaces were assessed quarterly. Campus A, with a history of higher financial expenditures, was monitored weekly to assess whether support surfaces selections met guideline recommendations. Nursing staff reviewed hospital protocol regarding guidelines before implementation, and a self-administered review test was required during the first year post-implementation. One year later, a modest decrease in annual expenditures for rental support surfaces was noted. Campus A had a decrease in nosocomial pressure ulcers, while Campus B had an increased prevalence rate. Staff selection of support surfaces, within guideline recommendations, improved to 75% on medical/surgical units, and 98.8% in ICUs on Campus A. Although implementing support surface selection guidelines did not result in a significant reduction in cost, it created a framework for monitoring future related decisions. PMID- 11889750 TI - Overcoming barriers to quality wound care: a systems perspective. AB - The societal burden of chronic wound care is guaranteed to intensify given the increasing elderly population of the United States. Currently 1.5 to 3 million patients with pressure ulcers are being treated at an estimated cost of $5 billion dollars annually. Many more millions suffer from venous, arterial, and neuropathic leg ulcers. When the huge scope of the issue is considered, the enormous challenge of providing quality wound care in the future is evident. Using general systems theory, this article analyzes contemporary salient barriers to quality chronic wound care from individual, group, and societal level systems perspectives. Factors that can help overcome these barriers are targeted, including newly emerging technological facilitators. PMID- 11889752 TI - By the numbers. PMID- 11889751 TI - The role of "wound burden" in determining the costs associated with wound care. AB - Because of the tremendous resources they require to heal, patients with severe wounds present significant challenges to our healthcare system. This study was undertaken to introduce the concept of "wound burden" and its predictive value in anticipating the costs associated with inpatient care for patients with wounds. Wound burden is a new concept that can be used to represent the severity of a patient's skin breakdown; it is defined in this study in terms of number, size, and stage. A computerized system of wound cost tracking measured the costs involved in delivering optimal wound care to 240 patients in a long-term acute care facility. Patients were stratified in a system that accounted for "wound burden" to determine the degree to which wound burden is related to costs. Costs that pertained to supplies, specialty beds, nutrition, labs, and extra personnel time required to document and care for the wounds were recorded. The concept of "wound burden" was presented and found to be very important in predicting actual costs. Patients with the highest level of wound burden were found to have significantly higher wound costs and total stay costs (P > 0.0001). As payment systems change, having data available to justify the resources necessary to allow facilities to continue to care for the most highly wound burdened patients will become increasingly important. PMID- 11889753 TI - Angiogenesis in necrotic ulcers treated with hyperbaric oxygen--comparing apples with apples. PMID- 11889754 TI - Definitions revisited. PMID- 11889755 TI - [Molecular neuropathology of JC virus]. PMID- 11889756 TI - [Recent progress in the research of multiple system atrophy with special references to alpha-synuclein and suprachiasmatic nucleus]. PMID- 11889757 TI - [Carotid ultrasonographic and brain computerized tomographic findings in patients with vascular ocular syndromes]. AB - To clarify the characteristics of cerebrovascular lesions in subtypes of vascular ocular syndrome, including amaurosis fugax(AF), retinal artery occlusion(RAO), and retinal vein occlusion(RVO), 93 patients with vascular ocular syndrome were studied by means of carotid ultrasonography(US) and brain computerized tomography(CT). The subjects comprised 21 patients with AF, 37 with RAO, and 35 with RVO who were sequentially given these diagnoses by the department of ophthalmology. On the basis of US findings, carotid lesions were defined as the presence of plaque or stenotic changes. CT findings were assessed for the presence and distribution of low-density areas(LDAs). Mean age was similar in each group, ranging from 64.5 to 67.4 years. The RAO group had high rates of men, hypertension, and smokers. US showed that the prevalence of carotid lesions ipsilateral to the affected eye was high in the RAO group and that severe stenosis and ulcerated plaque were present in 28.6% of the AF group and 45.9% of the RAO group. On CT examination, cerebral infarctions appeared as LDAs in about 10% of the patients in each group, and the incidence and distribution of LDAs were similar. Of 13 patients with cerebral infarction, only 2 were presumably due to carotid lesions; the others had a variety of causes. The discrepancy between US and CT findings was attributed to the small number of patients with cerebral infarction, since most patients had visual defects as an initial symptom. Our results suggest that extracranial carotid lesions, considered to be a major risk factor for stroke, should be carefully assessed in patients with AF or RAO to prevent further stroke. PMID- 11889758 TI - [Benefit of L-DOPA-without-DCI (decarboxylase inhibitor) therapy on wearing-off phenomenon in advanced stages of Parkinson's disease patients]. AB - Motor fluctuation is the most annoying complication experienced by patients in the advanced stages of Parkinson's disease. A Combination therapy of a dopamine receptor agonist and levodopa/DCI(DOPA-decarboxylase inhibitor) is commonly used to control the complication. Although administration of levodopa/DCI is useful in minimizing peripheral side effects of levodopa, it increases the incidence of motor complications due to the marked fluctuation of plasma levodopa level. The use of levodopa without DCI might be an option for controlling motor fluctuation, because the extent of plasma levodopa level fluctuation is smaller when levodopa is administered without DCI than with DCI. Six patients with Parkinson's disease who had troublesome motor complications under levodopa/DCI and DA agonist combination therapy were compared in terms of the extent of motor complications and their satisfaction after changing their therapy from levodopa/DCI to levodopa without DCI. The change from levodopa/DCI to levodopa(without DCI) was carried out all at once, and the levodopa/DCI to levodopa dose ratio was started at 1:4. The dose of levodopa(without DCI) was then increased gradually until motor complications improved or side effects were observed in patients. Except two patients who voluntarily quitted levodopa and restarted DOPA/DCI before the dose of levodopa fixed, all cases showed improvement of wearing-off phenomenon. No adverse event was observed. Levodopa-without-DCI-therapy was effective for controlling motor fluctuation in patients of Parkinson's disease in advanced stages. PMID- 11889759 TI - [Tandospirone citrate, a selective 5-HT1A agonist, alleviates L-DOPA-induced dyskinesia in patients with Parkinson's disease]. AB - A rapid and excessive increase in extracellular dopamine(DA) after L-DOPA administration is considered one of the major causes for L-DOPA-induced peak-dose dyskinesia. Therefore, inhibition of excessive rise in L-DOPA-derived DA is likely to be an ideal treatment for L-DOPA-induced dyskinesia. Based on our previous experimental studies that 8-OH-DPAT, a potent 5-HT1A agonist, attenuates an increase in L-DOPA-induced extracellular DA in the striatum of the rat model of Parkinson's disease, we hypothesized that L-DOPA-induced dyskinesia in patients with Parkinson's disease is alleviated by a 5-HT1A agonist. In the present study, we administered tandospirone citrate, a selective 5-HT1A agonist, to patients with Parkinson's disease suffering from L-DOPA-induced dyskinesia. Tandospirone(15-60 mg/day) was administered to 10 patients with L-DOPA-induced peak-dose dyskinesia. Twelve weeks after tandospirone treatment, duration of dyskinesia, subjective and objective severity of dyskinesia, and parkinsonian features were evaluated. Severity of dyskinesia was decreased in 5 patients; among these, 3 patients experienced slight worsening of parkinsonian features. Four patients showed no change in dyskinesia; among these, 2 patients showed worsening of parkinsonian features. One patient had slight worsening of dyskinesia without any change in parkinsonian features. The present study demonstrated that tandospirone is effective in alleviating L-DOPA-induced dyskinesia in 50% of the patients. However, at the same time 50% patients showed slight worsening of parkinsonian features. Both the anti-dyskinetic effect and the worsening of parkinsonian features are thought to be induced by tandospirone's potent 5-HT1A agonistic activity. Diverse effect of tandospirone may be caused by its partial agonist activity on 5-HT1A receptors, or may indicate that other causes for the expression of dyskinesia exist apart from excessive rise in brain DA levels. Administration of a 5-HT1A agonist is a choice for patients with dyskinesia if the care is taken so as not to induce worsening of parkinsonian features. Further studies such as double-blind trials are needed to confirm the usefulness of a 5-HT1A agonist for L-DOPA-induced dyskinesia. PMID- 11889760 TI - [Body temperature in the super-acute phase of the cerebrovascular disorders: mainly focused on the subarachnoid hemorrhage]. AB - BACKGROUNDS AND PURPOSE: Body temperature in the acute phase of cerebrovascular disorders(CVDs) may influence the outcome. However, the natural course of body temperature after CVDs has not yet been clarified. The purpose of this study was to elucidate the natural courses of body temperature after CVDs. PATIENTS AND METHODS: We retrospectively investigated 681 patients with CVDs(subarachnoid hemorrhage(SAH): 478, cerebral ischemia: 47, intracerebral hemorrhage(ICH): 156) who were admitted within 24 h after onset. The body temperature was measured with an electronic thermometer at the axilla on admission. The body temperatures of 73 patients with non-ruptured cerebral aneurysms on admission(admitted between 09:00 and 15:00) were used as normal control group. RESULTS: The body temperature in the control group was 36.49 +/- 0.45 degrees C. In comparison, the temperature in the SAH group was significantly lower(35.88 +/- 1.00 degrees C, n = 338, p < 0.001) when the patients were admitted within 4 h after onset, and significantly higher (36.80 +/- 0.85 degrees C, n = 140, p < 0.05) when they were admitted after 4 h and up to 24 h. There was a significant negative correlation between the severity of the SAH and body temperature within 4 h and a significant positive correlation beyond 4 h. Body temperature in the cerebral ishcemia group was significantly lower than in the control group(36.09 +/- 0.59 degrees C, n = 17, p < 0.05) when the patients were admitted within 2 h, but was close to that in the control group when they were admitted beyond 2 h and up to 24 h after onset (36.45 +/- 0.58 degrees C, n = 30). The falls of body temperature in the super-acute phase in the SAH and the cerebral ischemia groups were observed in patients admitted between 09:00 and 15:00. Although body temperature in the ICH group was slightly lower when the patients were admitted within 4 h and slightly higher when admitted beyond 4 h and up to 24 hours after onset, no significant differences were observed in comparison with the control group. In the super acute phase of the cerebral ischemia and the ICH, body temperature tended to be lower in the patients with worse condition. CONCLUSION: This study clearly demonstrated that body temperatures in patients with CVDs changed rapidly within 24 h after onset. Body temperature in the SAH group within 4 h and that in the cerebral ischemia group within 2 h after onset was significantly lower than in the control group. These temperature falls were not the products of circadian rhythm. The temperature in the SAH group beyond 4 h and up to 24 h after onset rose significantly. Comparison with normal controls and consideration of the circadian rhythm are important when studying changes of body temperature in patients with CVDs. PMID- 11889761 TI - [Giant fusiform aneurysm with dissection in the middle cerebral artery: a case report]. AB - A rare case of a giant fusiform aneurysm with dissection in the middle cerebral artery is reported. A 37-year-old man was referred to our hospital for severe headache and left retrobulbar pain. MRI and angiography showed a giant aneurysm in the temporal branch(M 2) of the left middle cerebral artery. We supposed that dissection had occurred from a giant fusiform aneurysm in the middle cerebral artery judging from presentation, MRI findings, angiography findings, and operative findings. Giant fusiform aneurysms in the middle cerebral artery(MCA) are uncommon cerebral aneurysms. The surgical approach or endovascular treatment to giant fusiform aneurysms in the MCA is technically difficult, so some patients are treated conservativery. However they have a poor natural history that differs from that of typical saccular aneurysms. And they have character of weakness in the internal elastic lamina, and therefore have the potential to dissect. We suggest that appropriate therapy including conservative therapy and surgical techniques is necessary for giant fusiform aneurysms even if they are found incidentally. PMID- 11889762 TI - [A case of relapsing myelitis associated with hypocomplementemia, presenting with Lhermitte sign enhanced by truncal flexion]. AB - A 36-year-old woman suffered from steroid-responsive relapsing myelitis associated with hypocomplementemia, thrombocytopenia, and anti-cardiolipin antibody. At the second attack of paraplegia, neck flexion in a supine position induced uncomfortable dysesthesia radiating into the ulnar side of the bilateral forearms. Both truncal and neck flexion resulted in painful dysesthesia down into both lower limbs also. Cervical MRI showed 2 gadolinium-enhanced dorsal-dominant lesions in the spinal cord at C 5/6 and C 6/7 disc levels. Enhancement of Lhermitte sign by truncal flexion might be useful to detect multiple lesions in the dorsal column. PMID- 11889763 TI - [A case of medial temporal lobe epilepsy associated with occult focal cortical dysplasia in the lateral temporal neocortex]. AB - A 29-year-old male with medial temporal lobe epilepsy(MTLE) was revealed to have "occult" focal cortical dysplasia(FCD) in the lateral temporal neocortex. He had no history of febrile convulsion and developed complex partial seizure at the age of 14 year, which became intractable. Although MRI failed to reveal structural abnormality in the temporal lobe, even retrospectively, the findings of non invasive preoperative examination, such as video-EEG monitoring and interictal ECD-SPECT and FDG-PET, were consistent with those of the left MTLE. Intraoperative electrocorticography(ECoG) demonstrated almost continuous paroxysmal activities on the anterior part of the inferior temporal gyrus(ITG). Anterior temporal lobectomy(ATL) with hippocampectomy was performed. Histological examination revealed FCD in the small area with 0.8 mm in diameter of the resected ITG. In the ATL without preoperative invasive examination such as chronic subdural electrode recording, intraoperative ECoG recording is mandatory. PMID- 11889764 TI - [Proton MRS in a case of intracerebellar epidermoid]. AB - We report a case of epidermoid, in which proton MR spectroscopy(MRS) provided additional information to MRI. The tumor revealed high signal intensity on T2 weighted images(WI) and low signal intensity on T1 WI. No enhancement was observed. Proton MRS was acquired with the Proton Regional Imaging of Metabolites(PRIME) method(TR/TE/measurements = 2,000 ms/136,272 ms/128 times). Proton MRS revealed lactate(Lac) peak at 1.33 ppm as negative peak at TE 136 ms and positive peak at TE 272 ms. N-acetylaspartate(NAA), creatine/phosphocreatine(Cr) and choline-containing compounds(Cho) were not visible. Lac peak without other peaks such as NAA, Cr, Cho is a unique finding of proton MRS for epidermoid and this could be useful in the differential diagnosis. PMID- 11889765 TI - [Agenesis of internal carotid artery: differences between arterial agenesis and occlusion]. PMID- 11889766 TI - [A sporadic case of dentatorubral-pallidoluysian atrophy diagnosed by MRI findings]. PMID- 11889768 TI - [Neural stem cells: their identification, isolation and potential therapeutic application]. PMID- 11889767 TI - [A 68-year-old woman with dementia and parkinsonism]. AB - We report a 68-year-old woman who developed progressive dementia and parkinsonism. She was well until 1990 when she was 58 years of age. She started to show memory loss. Four years later, she developed difficulty in dressing and behavioral problems such as eating rice with her hands, going out of her house without purposes, and difficulty in finding the rest room in her house. She was admitted to the neurology service of Hatsuishi Hospital on January 19, 1996, when she was 64 years of the age. On admission, she was alert but markedly demented. The score of Hansegawa Dementia Scale was 0/30. She was unable to make any coherent conversation. She appeared to have dressing apraxia but did not appear to have aphasia. Cranial nerves were intact. She walked in small steps with stooped posture. She did not have motor weakness but she showed plastic rigidity in all four limbs. No tremor or ataxia was noted. Deep tendon reflexes were within normal limits but the plantar response was extensor bilaterally. She continued to deteriorate after admission. In May of 1998, she started to fall. In June of 1998, she had a generalized convulsion. In January of 1999, she became unable to take foods orally and a gastrostomy was placed. She expired on May 29, 1990. She was discussed in a neurological CPC and the chief discussant arrived at the conclusion that the patient had Alzheimer's disease. The question was whether her parkinsonism was a part of her Alzheimer's disease or she had an additional disease to explain her parkinsonism. Post-mortem examination revealed moderate to marked atrophy of the frontal and the temporal lobes as well as in the limbic areas with dilatation of the lateral ventricles. Marked neuronal loss was noted in the CA 1 to the subiculum region with gliosis. Neurofibrillary tangles were seen in the remaining neurons. Neuropil threads were seen by Gallyas-Braak staining. Similar changes were seen in the parahippocampal gyrus and in the entorhinal cortex. Senile plaques were seen in the insular cortex and in other cortical areas. Cortical type Lewy bodies were seen in the cingulate cortex. The Meynert nucleus showed marked neuronal loss and gliosis. The substantia nigra and the locus coeruleus showed moderate loss of pigmented neurons. Lewy bodies were seen in these regions. The dorsal motor nucleus of the vagal nerve was retained, however, one Lewy body was observed. Pathologic diagnosis was Alzheimer's disease plus Parkinson's disease. It is an interesting question whether or not her parkinsonism was due to nigral lesion or frontal lesions. It is known that parkinsonism may complicate in advanced Alzheimer's disease not necessarily due to nigral lesion. On the other hand, in incidental Lewy body disease, the substantia nigra shows mild Parkinson's disease-like change without clinical parkinsonism. This patient appeared to have been a true complication of Alzheimer's disease and Parkinson's disease. PMID- 11889769 TI - [Diversity in molecular organization and function of plastid nucleoids--a hypothesis on the discontinuous evolution of plastid genomic machinery]. PMID- 11889770 TI - [Probing the structure and function of enzymes by using transition-state analogues]. PMID- 11889771 TI - [Structural biology of glycoproteins of immunological interest]. PMID- 11889772 TI - [CAAX peptidomimetics: their farnesyltransferase inhibition activity and antitumor effect]. PMID- 11889773 TI - [Energetics of enzymatic reactions]. PMID- 11889774 TI - [Toward observing chemical reactions of proteins with quantum chemistry]. PMID- 11889775 TI - [Effects of fentanyl on acetylcholine release from hippocampus and righting reflex in rat: an in vivo brain microdialysis study]. AB - Fentanyl, a synthetic mu-opioid agonist, is one of the most popular opioids in clinical anesthesia. It is well-known that memory is affected by opioids and opioid antagonists in animals. Cholinergic neurones releasing acetylcholine (ACh) particularly in the hippocampus have been shown to be related to memory. This study, therefore, was conducted to examine the effects of fentanyl on hippocampal ACh release in rats. The experiments were performed on male SD rats under freely moving condition by in vivo brain microdialysis technique. After taking initial three samples, test drugs were administered. The ACh release was determined by HPLC-ECD method. Effect on behavior was determined as loss of righting reflex (LRR). Fentanyl 50, 100 and 200 micrograms.kg-1 i.p. significantly decreased the ACh release from the rat hippocampus. Naloxone, a mu-antagonist, 100 micrograms.kg-1 i.p. antagonized the inhibitory effect of fentanyl 100 micrograms.kg-1 i.p. on the ACh release in the hippocampus. Only fentanyl 200 micrograms.kg-1 i.p. produced LRR. The effects of fentanyl, the dose of which is so little that it does not produce LRR, on the rat hippocampal ACh release may be controlled by the inhibitory actions of mu-opioid receptors. PMID- 11889776 TI - [Continuous epidural administration of droperidol to prevent postoperative nausea and vomiting]. AB - This randomized double-blind trial was designed to evaluate the antiemetic effect of continuous epidural analgesia with droperidol mixed with bupivacaine and buprenorphine. We studied 78 patients for abdominal gynecological surgery under general-epidural anesthesia. After recovery from anesthesia, they received epidural administration of 0.25% bupivacaine 40 ml and buprenorphine 0.4 mg with or without droperidol 2.5-5.0 mg at a rate of 2 ml.h-1 for 24 hours. The addition of droperidol 5.0 mg led to serious undesirable effects. Droperidol 2.5 mg, however, showed not only significant antiemetic effect without any adverse action, but also the reduction of rescue analgesics. We conclude that the addition of a small dose of droperidol to epidural analgesics reduces the incidence of postoperative emesis and the requirement of rescue analgesics. PMID- 11889777 TI - [The effects of prostaglandin E1 on systemic and cerebral oxygenation before and during cardiopulmonary bypass]. AB - The purpose of this study was to examine the effects of a small dose of prostaglandin E1 on systemic and cerebral oxygenation. Thirty patients for coronary artery bypass graft surgery were randomly divided into two groups: Group 1 received PGE1 25 ng.kg-1.min-1. Group 2 received PGE1 50 ng.kg-1.min-1. After measuring baseline hemodynamics and mixed (SvO2) and juglar (SjvO2) venous oxygen saturations, administration of PGE1 at a rate of 25 ng.kg-1.min-1 or 50 ng.kg 1.min-1 was started before and during CPB. In group 2, mean arterial pressure (MAP) decreased during CPB, while in group 1, MAP was unchanged during CPB. There was no change in SjvO2 both in group 1 and group 2 before and during CPB. The administration of PGE1 at a rate of 25 ng.kg-1.min-1 during CPB was suitable for the maintenance of SvO2 and SjvO2. PMID- 11889778 TI - [The effect of acupuncture or electro-acupuncture on circulatory parameters]. AB - We examined the effect of acupuncture or electro-acupuncture on circulatory parameters by use of a non-invasive procedure in 10 healthy male adult volunteers. Circulatory parameters measured were systolic blood pressure, pulse rate, total peripheral resistance and cardiac output. Acupuncture and electro acupuncture significantly increased total peripheral resistance, and electro acupuncture increased systolic blood pressure further. There was no difference in cardiac output compared with the control. The results suggest that acupuncture or electro-acupuncture has an action of alpha-stimulation but not of beta stimulation, and that electro-acupuncture has a stronger effect than acupuncture alone. PMID- 11889779 TI - [Pre-treatment with ketamine reduces incidence and severity of pain on propofol injection]. AB - The purpose of this study was to evaluate the effect of pre-treatment with ketamine on the reduction of pain during injection of propofol in adult patients. We conducted a prospective, randomized, double-blinded trial. Forty-three patients were randomly allocated to one of two groups according to the agents administered before hand; Group C, normal saline 2 ml and Group K, 1% ketamine 2 ml. The pain on injection was rated as none, mild, moderate, or severe. Sixty eight percent of patients in the C group experienced pain, while 33% of patients experienced pain in the K group. Thirty-six percent of patients in the C group complained moderate to severe pain but only 9% of patients in the K group. The mechanisms of prevention by ketamine of the pain on propofol-injection could not be clarified from our study, but it may be related to central effects of ketamine. In conclusion, ketamine pre-treatment before propofol administration significantly reduces incidence and severity of pain associated with propofol injection. PMID- 11889780 TI - [Venoarterial bypass in children with congenital heart disease--a retrospective study]. AB - To investigate prognostic factors for patients supported by venoarterial (VA) bypass, we analyzed retrospectively 31 patients with congenital heart diseases supported by VA bypass between 1997 and 2000. Median age was 3.6 months and median body weight was 4.0 kg. Causes of VA bypass were difficult weaning from cardiopulmonary bypass in 8 patients, postoperative cardiac dysfunction in 5 and cardiopulmonary resuscitation in 18. Six (19.4%) of 31 patients were weaned successfully from VA bypass, and 2 (6.5%) survived to hospital discharge. In patients who were weaned from VA bypass successfully, VA bypass was instituted more quickly (41.6 +/- 5.0 vs 62.5 +/- 35.9 min), and good tissue perfusion was established in association with lower lactate levels at 12 hr (7.0 +/- 5.9 vs. 16.4 +/- 15.4 mmol.l-1) and larger urine output during first 24 h (81 +/- 68 vs. 22 +/- 43 ml.kg-1.day-1), compared to those who failed to be weaned. Major complications were intracranial hemorrhage in the newborn (50%), and hypoxic brain damage in patients with palliative operation (35%). It is necessary to establish guidelines of entry, weaning, and withholding of this support. PMID- 11889781 TI - [Hemoglobin levels and weight gain after coronary bypass grafting by use of intraoperative hemodilution and autologous blood transfusion]. AB - We investigated the relationship between the degree of hemodilution during cardiopulmonary bypass (CPB) and weight gain after coronary bypass grafting by use of intraoperative hemodilution and autologous blood transfusion. There is no significant difference in weight gain between the higher hemoglobin group (Hb > or = 6 g.dl-1) and the lower hemoglobin group (Hb < 6 g.dl-1). Furthermore, there is no significant correlation between hemoglobin levels during CPB and weight gain after operation. We conclude that slightly excessive hemodilution than usual during CPB does not influence weight gain when cardiac and renal functions are fair. PMID- 11889782 TI - [Bilateral parotid gland swelling with hyperamylasemia following cesarean section under regional anesthesia--report of two cases]. AB - Bilateral parotid gland swelling with hyperamylasemia occurred in two patients on the first postoperative day after cesarean section under combined epidural-spinal anesthesia. Total ephedrine doses used to treat hypotension due to anesthesia were 48 and 52 mg, respectively. These symptoms and findings seemed to have been caused by beta stimulating effect of high dose ephedrine. PMID- 11889783 TI - [The changes in hemodynamics and dose requirements in total intravenous anesthesia using propofol and buprenorphine]. AB - A retrospective study was performed to evaluate the changes in hemodynamics and dose requirements in total intravenous anesthesia (TIVA) using propofol and buprenorphine without (Group S: spinal surgery (3-6 h), n = 8, 28-79 Y) or with (Group A: abdominal surgery (5-10 h), n = 15, 36-83 Y) epidural anesthesia. All patients were premedicated with midazolam i.m. (2-5 mg). Anesthesia was maintained with a single dose of buprenorphine (Group S: 1.9 +/- 0.4 micrograms.kg-1, Group A: 2.0 +/- 0.5 micrograms.kg-1), propofol infusion and vecuronium with 40% oxygen in air. Group A was supplemented with continuous epidural anesthesia using 2% mepivacaine. In Group A, mean arterial pressure (MAP) and heart rate remained stable after the start of surgery. However, in Group S, 2 hours after the start of surgery MAP increased (P < 0.05) and remained elevated (P < 0.05) at higher levels than those in Group A. The maintenance dose of propofol in Group A (4.0 +/- 1.1 mg.kg-1.h-1) was significantly smaller than in Group S (6.5 +/- 0.9 mg.kg-1.h-1). In both groups, infusion rates of propofol were unchanged from 1 hour after the start to the end of surgery. Infusion rates of mepivacaine (5.2 +/- 0.9 ml.h-1) were unchanged following the increase 2 hours after the start of surgery. Awakening times were within 25 min (Group S 11.3 +/- 7.2 min vs Group A 14.7 +/- 7.3 min). There was no awareness during anesthesia in either group. The results suggest that additional continuous epidural anesthesia in TIVA would be useful to reduce propofol dose, to stabilize hemodynamic state and to obtain rapid recovery in anesthesia of long duration. PMID- 11889784 TI - [Anesthetic management of four pediatric patients with CCAM for pulmonary lobectomy]. AB - We report perioperative management of four pediatric patients with congenital cystic adenomatoid malformation (CCAM) for pulmonary lobectomy under general anesthesia combined with thoracic epidural anesthesia. The patients were two newborn infants and two infants. A 23-day-old girl whose cyst contained air and a one-day-old girl whose cyst content was drained percutaneously before induction, showed uneventful courses during perioperative period. In a one-yr-old boy with a cyst filled with purulent material, the endotracheal tube became nearly occluded with copious purulent material during operative period, and the management of this case was extremely difficult. After this case, in the other infant case expected to have purulent cyst as the result of repetitive infection, we inserted 5 Fr Fogaty catheter into the orifice of the left inferior bronchus to protect the normal side. This case showed uneventful course during anesthesia. We conclude that anesthesiologists should choose proper airway management method depending on the nature of cystic fluid in these patients. PMID- 11889785 TI - [A case of suspected cholesterol embolism]. AB - A 53 year old male was admitted with the diagnosis of brain stem infarction. Severe stenosis of the bilateral internal carotid arteries was revealed by cerebral angiography. During the angiography, he complained of acute abdominal pain and was referred to abdominal surgical department. He was diagnosed as thrombo-embolism of the superior mesenteric artery and treated conservatively. The symptoms improved, but renal function became worse and the toe was found to be blue. Cholesterol embolism was suspected by nephrologists and thrombolytic therapy was discontinued. Steroid was administrated and LDL-apheresis was performed. The renal function was aggravating and hemodialysis was required. Emergency neurosurgical operations are sometimes performed immediately after cerebral angiography and the neuroanesthesiologists should be aware of the pathology of cholesterol embolism in perioperative management. PMID- 11889786 TI - [Anesthesia for a patient with xeroderma pigmentosum]. AB - The anesthetic management of a patient with xeroderma pigmentosum is described. A 17-year-old woman underwent tracheostomy because of progressive recurrent nerve palsy. The operation was performed uneventfully under general anesthesia using propofol and fentanyl. Xeroderma pigmentosum (XP) is an autosomal recessive disease that is characterized by hypersensitivity to sunlight with a high incidence of skin cancer and exhibits variable neurological abnormalities. Classical types of XP have a defect in nucleotide excision repair (NER). It has been reported that volatile anesthetics such as halothane deranged NER in cells obtained from an XP patient. Thus, general anesthesia using volatile agents should be avoided, if possible, because inhalation anesthetics may worsen the symptoms of XP. PMID- 11889787 TI - [Usefulness of multi-lead electrocardiogram and transesophageal echocardiography for the detection and evaluation of intraoperative coronary spasm]. AB - We describe a case of coronary spasm in a 59-year-old man undergoing an emergent abdominal aortic replacement for ruptured aortic aneurysm. The patient was brought to the operating room in a state of hypovolemic shock, and was successfully resuscitated through intensive volume expansion by rapid infusion devices. Twenty minutes after cross-clamping of the abdominal aorta, ST-segment elevation on the lead III of electrocardiogram (ECG) and dyskinesis in the inferior wall shown by transesophageal echocardiography (TEE) were noted. Coronary spasm was suspected, and isosorbide dinitrate was administered intravenously without delay, leading to prompt reversal of ischemic changes. A number of reports have suggested that care should be taken against coronary spasm in non-cardiac surgery as well as cardiac surgery, especially in patients with coronary risk factors. Monitoring by multi-lead ECG and TEE is a powerful method by which to detect and evaluate intraoperative myocardial ischemia. PMID- 11889788 TI - [Anesthetic management of MIDCAB with high dose diltiazem]. AB - Precise management of blood pressure and heart rate is required during minimally invasive direct coronary artery bypass (MIDCAB). Previously, low dose diltiazem and beta-blokers have been employed for control of circulation during this procedure, but we report 2 patients whose blood pressure and heart rate were managed during MIDCAB by high-dose diltiazem. In both patients, anesthesia was induced with propofol, vecuronium and fentanyl, and maintained by continuous infusion of propofol and inhalation of oxygen and nitrous oxide. Fentanyl, midazolam, and sevoflurane were administered occasionally. Immediately after the induction, a continuous infusion of nicorandil (2 to 4 mg.hr-1) was started and diltiazem (4 to 15 micrograms.kg-1.min-1) was administered continuously from the beginning of the surgery. Following discontinuation of diltiazem administration, blood pressure and heart rate returned to their preoperative values. These results suggest that safe anesthetic management during MIDCAB can be performed with highdose diltiazem. PMID- 11889789 TI - [Suddenly developing low output syndrome during acute thoracic aortic dissection surgery in a patient with progressive systemic sclerosis]. AB - A 70-year-old man with progressive systemic sclerosis (PSS) suddenly developed low output syndrome (LOS) after weaning from cardiopulmonary bypass during acute thoracic aortic dissection and died in an early postoperative stage because of multiple organ failure. PSS was kept in a relatively good condition before onset of his surgical disease, but his Raynaud's phenomenon was much worse and inflammation findings were very severe before his operation. We suspected that PSS might have been involved in LOS in this case. PSS has many cardiac complications, for example spasm or organic changes of coronary artery, cardiac fibrosis, arrhythmia, secondary cardiac failure of pulmonary hypertension, pericardial inflammation, and so on, but we speculate that LOS might be caused by peripheral vessel disturbance. LOS occurs easily in chronic hypovolemic states and abnormal response of peripheral vessels due to PSS. We advise more careful anesthetic and circulatory management especially in invasive operation in PSS patients. PMID- 11889790 TI - [Efficacy of propofol in controlling myoclonus during rewarming in a brain hypothermia patient]. AB - Propofol is an intravenous anaesthetic agent having anticonvulsant property. We report here a case in which propofol was effective in controlling myoclonus during rewarming in brain hypothermia patient. A 35-year-old male patient was admitted in a comatose state with right-sided hemiparesis, anisocoria and absence of bilateral light reflex. On admission, a head CT showed traumatic subarachnoid hemorrhage, left subdural hematoma, 10 mm midline shift and tentorial herniation with massive brain swelling together with extensive hypodensity in the frontal, temporal and occipital lobes bilaterally. Left decompressive hemicraniectomy, removal of hematoma and brain hypothermia therapy were started immediately. Postoperative head CT showed 15 mm midline shift. The temperature of the jugular bulb was maintained at 34 degrees C for 2 days together with sedation using midazolam under artificial ventilation. The patient was gradually rewarmed at a rate of 0.5 degree C per day from the third hospital day. Myoclonus of sudden onset developed on the patient's head and upper extremities on the third hospital day. An intravenous bolus injection of 10 mg midazolam and continued intravenous infusion of midazolam were given but they did not completely stop myoclonus. A bolus of propofol 60 mg was given intravenously and continuous intravenous infusion of propofol 2 mg.kg-1.hr-1 was started after which the progression of myoclonus disappeared. Myoclonus was kept controlled until the continuous intravenous infusion of midazolam and propofol was discontinued on the sixth hospital day, after which myoclonus occurred again after extubation on the seventh hospital day. The clinical course of this case suggests that propofol might be an alternative effective agent to suppress refractory myoclonus. PMID- 11889791 TI - [Sevoflurane can induce rhabdomyolysis in Duchenne's muscular dystrophy]. AB - Rhabdomyolysis is one of the perioperative complications in patients with Duchenne's muscular dystrophy (DMD). It has been suggested that sevoflurane can be used safely for anesthesia in patients with DMD. In this report, we describe a case with DMD who received anesthesia with sevoflurane, in which rhabdomyolysis developed postoperatively. A 6-year-old boy diagnosed as DMD was scheduled for tonsillectomy under general anesthesia. Preoperative laboratory examination revealed a high level of creatine kinase (CK) (16,000-32,000 IU.l-1). An abnormality of the dystrophin gene was detected by DNA analysis. Anesthesia was induced with sevoflurane without muscle relaxant, and maintained with sevoflurane in nitrous oxide and oxygen under controlled ventilation. The course of anesthesia was uneventful and the patient recovered smoothly. Three hours postoperatively, dark red urine with a high concentration of myoglobin (1,390,000 ng.ml-1) was recognized with a high level of CK (63,500 IU.l-1). Body temperature was 37.6 degrees C, and electrocardiogram and serum potassium were within normal ranges. After the diuresis with mannitol and furosemide, the urine became clear. On the 4th postoperative day, he was discharged without any complication. This case suggested that rhabdomyolysis can develop after sevoflurane anesthesia in patients with DMD. PMID- 11889792 TI - [A case of severe septicemia due to Aeromonas hydrophila]. AB - Septicemia by Aeromonas hydrophila (A. hydrophila) developed in a 69-year-old female with a history of uncompensated liver cirrhosis. She was admitted to our hospital, complaining of fever, vomiting, diarrhea, and pain as well as swelling of the left lower extremity. Seven hours later, skin of the left extremity developed bullae and became discolored. Although she was treated with infusion of antibiotics plus dopamine, continuous hemodiafiltration, glucose-insulin therapy and mechanical ventilation, she fell in severe septic shock, and died 23 hours after admission. A. hydrophila was isolated from both blood and bullous fluid. Mortality rate of septicemia due to A. hydrophila is reported ranging from 29% to 73%. Patients with liver disease have poor prognosis. Morbidity of this septicemia depends on pre-infection conditions, especially on liver functions. Early surgical debridement and antimicrobial therapy is recommended, but rapid worsening often results in death. We have to pay attention to infection of A. hydrophila especially in patients with liver cirrhosis. PMID- 11889793 TI - [Propofol anesthesia for a patient with progressive muscular dystrophy]. AB - We gave propofol anesthesia to a patient with limb-girdle type of progressive muscular dystrophy. A 42 year-old male was to have skin graft for third degree burn. His respiratory function test showed %VC of 73.6% and %FEV1.0 of 107.6%. Arterial blood gas data were within normal ranges. He was anesthetized with propofol, fentanyl, vecuronium and nitrous oxide. During position change, Wenckebach type of second degree AV block occurred. AV block returned to sinus rhythm easily by injection of ephedrine hydrochloride and atropine sulfate, and reduction of propofol infusion rate. There were no perioperative respiratory complications and no clinical manifestations of malignant hyperthermia. Propofol anesthesia is suitable for limb-girdle type of progressive muscular dystrophy, because of very little possibility of triggering malignant hyperthermia, rapid awaking, minimal residual effects of the respiratory system, and easiness in controlling anesthetic depth. PMID- 11889794 TI - [A case of successful thrombolysis by recombinant tissue plasminogen activator for postoperative pulmonary thromboembolism]. AB - A 52-year-old female suspected of hypercoagulability underwent modified radical hysterectomy and left oophorectomy for uterus cancer and left giant ovarian tumor under general combined with epidural anesthesia. On the day after the operation, the patient complained of dyspnea and developed tachypnea, a low Spo2, and hypotension after the intermittent external pneumatic compression of the legs. Echocardiography showed acute right cardiac failure and pulmonary angiography revealed massive pulmonary thromboembolism. The patient fell into shock with severe hypotension and unconsciousness during the catheter fragmentation and aspiration therapy for pulmonary thrombi. Bolus intravenous injection of monteplase 1.6 million units, a mutant of tissue plasminogen activator with a longer half-life, rapidly improved the shock status and stabilized the hemodynamic condition. Monteplase would be useful for life-threatening pulmonary thromboembolism although the risk of hemorrhagic complication remains. PMID- 11889795 TI - [A case of ventricular fibrillation during emergency clipping operation for cerebral aneurysm]. AB - A 59 year-old woman with subarachnoid hemorrhage underwent emergency neck clipping of cerebral aneurysm. Her preoperative examination showed atrial fibrillation, pulmonary edema and hypokalemia. Ventricular fibrillation developed immediately after clipping of the aneurysm and recurred 7 times thereafter during the surgery. Hypokalemia was corrected, and hypoxemia and other factors leading to ventricular fibrillation were excluded. RR interval was prolonged prior to ventricular fibrillation. Therefore intravenous temporary cardiac pacemaker was inserted immediately after the end of the surgery. It prevented successfully the prolongation of RR interval as well as ventricular fibrillation. The present case suggests that we should pay attention to the possibility of ventricular fibrillation during emergency radical surgery for ruptured cerebral aneurysm, and that cardiac pacemaker is useful to prevent ventricular fibrillation following prolongation of RR interval. PMID- 11889796 TI - [Anthocyanin pigments of flowers]. PMID- 11889797 TI - [Genes for flower coloration]. PMID- 11889798 TI - [Recent advances on reaction mechanism of anthocyanin biosynthesis]. PMID- 11889799 TI - [Biotechnology of flower color modification]. PMID- 11889800 TI - [Unraveling molecular mechanism of aerobic respiration in Escherichia coli]. PMID- 11889801 TI - [Molecular mechanisms of the intracellular localizations of Ca2+/calmodulin dependent protein kinase II isoforms, and their physiological functions]. PMID- 11889802 TI - [The mechanism of discontinuous replication: how it was established]. PMID- 11889803 TI - [Finding genes from genome sequences]. PMID- 11889804 TI - [Clinical study of double primary cancer involving the lung in resected cases]. AB - Among all cases of surgically resected lung cancer, there were 56 cases (16.1%) of double primary cancer. The common sites of the other primary cancer was the stomach (19 cases), followed by large intestine (9 cases), urinary bladder (7 cases) and pharinx-larynx (7 cases). One patient had triple cancers. In all cases of double primary cancer, 46 cases were metachronous, 10 of which were cases of initial lung cancer. The 5-year survival rate of double primary cancer was 39.7%. Good result was obtained in metachronous cases with initial lung cancer. Most of prognosis of double primary cancer was determined by that of lung cancer. In more than half of initial cancer, the second primary cancer was detected by symptoms. So, special attention to the possibility of double primary cancer in patients with resected lung cancer is necessary for improvement of prognosis. PMID- 11889805 TI - [Aortic arch replacement with covered stent-graft as elephant trunk for a type A acute aortic dissection]. AB - We report a case of aortic arch replacement with a covered stent-graft as an "elephant trunk". A 54-year-old woman was diagnosed with Stanford type A aortic dissection. The initial intimal tear was located in the distal aortic arch. Under deep hypothermic circulatory arrest and retrograde cerebral perfusion, the distal end of the arch graft, which was turned inside out and reinforced with a Z-stent, was inserted into the distal true lumen as an "elephant trunk". Distal anastomosis was performed between the aortic wall and the inverted external graft. Graft replacement of the aortic arch and ascending aorta was followed by proximal arch grafting. Coronary artery bypass grafting to RCA was performed concomitantly. The postoperative course was uneventful, and the distal false lumen became thrombosed. This procedure is effective for reliable distal anastomosis and prevention of blood leakage into the distal false lumen. PMID- 11889806 TI - [Bronchoscopic treatment for upper tracheal lesions with a laryngeal mask]. AB - A laryngeal mask provides maintaining airway with a larger inner diameter of the tube. A little information is available about bronchoscopic treatment for upper tracheal lesions. Three patients undergoing bronchoscopic treatment for upper tracheal lesions with a laryngeal mask were reviewed. The patients include 3 women, having 2 thyroid cancers and 1 thyroid goiter. The aims of the procedure were hemostasis and reduction of the tumor with subsequent endotracheal stenting in 2 patients, and endotracheal stenting in one patient. The treatment was performed under general anesthesia using a laryngeal mask. All cases were successfully treated without operative and postoperative complications related to the use of the laryngeal mask placement. Use of a laryngeal mask may facilitate insertion and retrieval of a flexible bronchoscope and instruments with an excellent manipulation in therapeutic bronchoscopy for subglottic and upper tracheal lesions. PMID- 11889807 TI - [Veno-venous ECMO for the respiratory failure after coronary artery bypass grafting]. AB - We encountered a 60-year-old man with severe respiratory failure after CABG. The patient received veno-venous extracorporeal membrane oxygenation (ECMO) in which blood was drained from the right atrium through a cannula and was returned via a cannula in the left femoral vein. During ECMO, the flow rate was 2.8-3.0 l/min (about 40 ml/kg/min) and the assist was needed for 5 years. Veno-venous ECMO is a simple and effective therapy for acute respiratory failure unresponsive to conventional treatment. PMID- 11889808 TI - [Is minimal skin incision and partial sternotomy approach for congenital heart defects less invasive?; evaluation of SIRS on ventricular septal defect]. AB - OBJECTIVES: Minimally invasive cardiac surgery (MICS) has been developed to offer patients the benefits of open heart operations with limited skin incision. But it is unclear whether this procedure is less invasive. We evaluate postoperative duration of systemic inflammatory response syndrome (SIRS) on ventricular septal defect (VSD). METHODS: From August 1997 to March 2000, 47 patients VSD underwent total repair by the minimal skin incision and lower partial median sternotomy (MICS group). We compared duration of SIRS between MICS and conventional method group (n = 14) and between early MICS and recent MICS group. We also evaluated the relationship between MICS and postoperative clinical course. RESULTS: Duration of SIRS of MICS group were obviously shorter than that of conventional method group (p < 0.05). That of recent MICS group is also significantly shorter than that of early MICS group (p < 0.05). Operative time, bypass time, postoperative intubation time and length of skin incision were related duration of SIRS. CONCLUSIONS: The results of this study indicate that MICS for VSD may be less invasive method. PMID- 11889809 TI - [Delayed hydrothorax induced by a pericutaneous central venous catheter; report of a case]. AB - We report herein a case of 53-year-old woman who suffered a hydrothorax induced by a central venous catheter which had been placed to facilitate total parenteral nutrition. The central venous catheter was inserted into the superior vena cava through the right subclavian vein. Chest X-ray film after insertion revealed proper position of the tip. She suddenly developed dyspnea and tachycardia due to right-sided hydrothorax 21 days after the insertion of the catheter. Chest X-ray showed massive pleural effusion in the right thorax, and the catheter tip inadvertently turned upward. The continuous mechanical force of the catheter tip against the SVC wall was considered to be the cause of this life-threatening delayed hydrothorax. PMID- 11889810 TI - [A case of emergency coronary artery bypass graft for the patient using chronic hemodialysis, where in blood volume was measurable in the perioperative period]. AB - An emergency coronary artery bypass graft (CABG) was given to a 66 year-old patient due to acute myocardial infarction (AMI). Circulating blood volume (BV) was measured to study in the perioperative period. Three coronary artery bypasses were made under cardiopulmonary bypass, being managed by ultrafiltration when the pump-oxygenator was in action and by peritoneal dialysis in the early postoperative period. Preoperative BV reduced immediately after the operation. It showed an increasing trend 4 hours after the operation, but after that BV reduced from that before the operation while water balance was kept positive. Cardiac output after the operation was higher than before. It suggested that in this patient using hemodialysis BV levels turned to be lower compared with that before the operation, as excessive water leaked out of the blood vessel, although water balance was kept positive due to improved cardiac functions after the operation. PMID- 11889811 TI - [Combined surgery for cardiovascular disease and general thoracic lesions]. AB - Surgical management of patients with concomitant critical cardiovascular disease and resectable general thoracic lesions is controversial. During a 16-year period (1985 to 2001), 15 patients underwent combined cardiovascular and general thoracic operations, of the 2,459 patients who underwent a cardiovascular operation requiring cardiopulmonary bypass at our institution. Patients had cardiovascular symptoms only and the general thoracic lesions were incidentally found by preoperative chest roentgenograms and/or computed tomography. Because of the cardiovascular disease, a pathological diagnosis was precluded before surgery. All except one descending thoracic aortic operation underwent concurrent pulmonary resection after neutralization of protamine following cardiovascular surgery requiring extracorporeal circulation. Lung pathology consisted of pulmonary bullae (n = 7), primary lung cancer (n = 4), benign lung tumor (n = 2), metastatic lung cancer (n = 1), and thymic cyst (n = 1). The pulmonary operations include bullectomy (n = 7), wedge resection (n = 6), lobectomy (n = 3), and removal of a thymic cyst (n = 1) including 2 staged procedures. The final diagnoses in 4 lung cancer cases were T1. N0M0, stage IA (n = 3) and T2N2M0, stage IIIA (n = 1). All malignancies including metastatic lung cancer, were able to be completely resected. The mean intraoperative bleeding volume for the cases was 997 +/- 221 ml, while mean duration of surgery was 382 +/- 31 minutes. Except for 2 cases required long term ventilatory support, the mean durations of tracheal intubation and ICU stay were 2.2 +/- 0.2 and 3.8 +/- 1.0 days respectively. Except for 1 surgical death, mean survival duration and 5-year survival rate were 59.7 +/- 12.5 (5-177) months and 66.3% respectively. These findings suggest that combined pulmonary resection with cardiovascular surgery is safe and offers a favorable prognosis to a selected group of patients. PMID- 11889812 TI - [The treatment for the acute occlusion of a coronary artery by PTCA failure with the acute occlusion of a femoral artery by the IABP insertion]. AB - A 68-year-old woman had the acute occlusion of a coronary artery by PTCA failure acute occlusion of a femoral artery by the IABP insertion. The patient underwent on pump beating coronary bypass and axillo-femoral bypass simultaneously. PMID- 11889813 TI - [Esophagopericardial fistula occurring at the 22nd years after operation for achalasia presenting as myopericarditis]. AB - We describe a case of pneumopericardium that resulted from esophagopericardial fistula occurring at the 22nd years after operation for achalasia in a 52-year old male. Atypical features on initial examination suggested myopericarditis or congestive heart failure. A chest roentgenogram and computed tomography revealed development of pneumopericardium. Subsequent emergent pericardiocentesis relieved cardiac tamponade and enabled us to diagnose pyopneumopericardium and esophagopericardial fistula in postoperative period. The esophagopericardial fistula was surgically closed. However, the patient died 2 weeks postoperation of hemorrhage from duodenal ulcer. Autopsy confirmed the pre-mortem diagnosis of esophagopericardial fistula without evidence of malignancy. Early diagnosis and subsequent treatment appears to be most important. This case report reinforces the difficulty of this diagnosis and perhaps the need for clinical awareness and inclusion of this entity in a differential diagnosis. PMID- 11889815 TI - [A case of mitral regurgitation with a giant thrombus thought to be caused by anti-phospholipid syndrome]. AB - A 58-year-old woman suffered from mitral valve regurgitation with a giant thrombus in right atrium was thought to be complicated by anti-phospholipid syndrome (APS). Positive anti-loops anti-coagulant and histories of cerebral infarction and miscarriage strongly indicated the diagnosis of APS. She underwent mitral valve replacement using bileaflet mechanical prosthetic value and removal of thrombus. She recovered uneventfully and strict anti-coagulation therapy has been continuing. PMID- 11889814 TI - [A case report of ross operation and ventricular septal defect closure following correction of type A interruption by modified Blalock-Park, pulmonary artery banding and patent ductus arteriosus division]. AB - A 2-year-old boy who had undergone a correction of a type A interruption using a modified Blalock-Park operation, pulmonary artery banding and the division of a patent ductus arteriosus, underwent a Ross operation and closure of ventricular septal defect (VSD). Although a pre-operative echo cardiogram revealed a bicuspid aortic valve, and a Doppler echocardiogram showed only 10 mmHg of pressure gradient across the aortic valve, Ross procedure was performed as a procedure accompanying the closure of a total conus VSD. The total conus VSD was closed with a Dacron patch using pledget mattress sutures. In addition, a running suture was applied over the denuded aortic root and the cranial margin to achieve water tight closure. An aortic root replacement procedure was our first choice for the Ross operation. After both coronary buttons were re-implanted into pulmonary sinuses, a pulmonary artery autograft was wrapped around by the remaining aortic wall for reinforcement to prevent future dilatation. The main pulmonary artery was reconstructed using a bicuspid pericardial valve conduit with a diameter of 24 mm. A post-operative echocardiogram showed no neoaortic valve regurgitation, good coaptation of tri-leaflets, mild regurgitation of pericardial valve and good cardiac performance. PMID- 11889816 TI - [MIDCAB for redo patients using the LITA-RA composite graft]. AB - Redo coronary artery bypass grafting (CABG) is associated with higher mortality, low-output syndrome, perioperative myocardial infarction than primary CABG. Minimally invasive direct coronary artery bypass grafting (MIDCAB) technique avoids the manipulation of old graft and injury of the adhesive heart in redo operation. We performed the MIDCAB procedure for 2 redo cases using the left internal thoracic artery (LITA)-radial artery (RA) composite graft. The LITA-RA composite graft was anastomosed to the left anterior descending branch (LAD) through small left anterior thoracotomy without cardiopulmonary bypass. Postoperative coronary artery graphy shows the widely patent of new graft. The MIDCAB procedure using the LITA-RA composite graft is safe and useful to regulate the bypass graft length and avoid the widely harvest of LITA in redo operation. PMID- 11889817 TI - [A modified Fontan operation in the presence of a supracardiac total anomalous pulmonary venous connection]. AB - We report a case of 2-year-old girl with asplenia syndrome who successfully underwent modified Fontan procedure and concomitant repair of supracardiac total anomalous pulmonary venous connection (TAPVC). The preoperative diagnosis included a common atrioventricular canal (type C), a double outlet right ventricle, a common atrium, common atrioventricular valve regurgitation, pulmonary stenosis, and a bilateral superior vena cava (SVC). Cardiac catheterization revealed a Qp/Qs of 1.3, mean PA pressure of 16 mmHg and an Rp of 1.3. The TAPVC drained to left SVC (LSVC) at a position proximal to the hemiazygos vein with an ostium of 5 mm in diameter. The LSVC was divided distal to its connection to the common pulmonary vein (CPV). The TAPVC ostium was cut back into the CPV, then it was anastmosed with posterior aspect of the atrial wall in an effort to provide a wide anastomosis. The postoperative course was uneventful and the patient was discharged from hospital on the 35th postoperative day. PMID- 11889818 TI - [Congenital tracheoesophageal fistula; report of an adult case]. AB - We experienced an adult case of congenital tracheoesophageal fistula which was successfully treated by surgical intervention. A 69-year-old female was admitted with complaint of coughing and fever. The patient previously had had several episodes of pneumonia. Esophagography and thoracic CT demonstrated a tracheoesophageal fistula in the upper of the thoracic esophagus. Following the resection of the fistula, the trachea was closed with one layer suture and the esophagus was closed with two layer sutures. A pedicled fifth intercostal muscle was interposed between the tracheal and esophageal suture lines. There was mild inflammatory change in surrounding tissues of the fistula. Pathological examination revealed that the fistula was covered with the squamous epithelium, and had the muscularis mucosa. Her postoperative course was uneventful. PMID- 11889819 TI - [A case of bronchioloalveolar carcinoma diagnosed by a segmental resection]. AB - A 66-year-old man who underwent an extirpation of thymoma in stage I on September 16 1997 was followed in the outpatient clinic. In October 1998, a chest CT scan revealed a 2.0 x 1.0 cm faint frosted glass like shadow in the right S9. On September 13 1999, the patient was admitted to the hospital for close examination. Two times of transbronchial lung biopsy could not offer any clear diagnosis because the lesion was present so as to encircle the central segment of the B9 gronchus. A segmental resection (S9 + 10) under thoracotomy was performed. Intraoperative frozen section diagnosis was bronchioloalveolar carcinoma (type A according to Noguchi's classification), and a resection of the remnant lower lobe and mediastinal lymph nodes dissection (ND 2 a) were added. With expected increase in frequency of detecting early pulmonary cancer's through CT, clinical cases for which we are obliged to diagnosed the disease by segmental resection may increase, if the lesion develops in the vicinity of the hilum of lung like this case. PMID- 11889820 TI - [Three cases with a mediastinal neurinoma developed in a same family]. AB - A 47-years-old woman (case 1) was admitted to our hospital because of a mediastinal mass. We performed an operation by VATS under the diagnosis of a mediastinal neurinoma. The histology of it was a neurinoma. Her father (case 2) had undergone a resection of a mediastinal neurinoma at the age of 42. Her brother (case 3) also had undergone a resection of mediastinal and intrathoracic neurinomas at the age of 37. A few years later, he underwent operations for neurinomas in limbs 2 times. We suppose patients with a mediastinal neurinoma have little complaints in many cases, so there are a number of patients who have a mediastinal neurinoma without being discovered and treated. Although the neurinoma is not considered as a hereditary disease inherently, the cases, we experienced, might have some genetic disorders. In this concern, our cases are very rare and have a great interest. PMID- 11889821 TI - [Stress management in European countries and US]. AB - In recent years, job stress and stress management have emerged as key issues in health promotion in the workplace in all of the post-industrialized countries. In the EU, the European Survey on Working Conditions has reveals that stress and musculo-skeletal disorders are the main health risks at work. In the US, NORA (national occupational research agenda) identifies 21 research priorities, in which "organization of work" is included as a job stress related factor. In this paper, trends and characteristics in occupational stress management in western countries, especially in the EU are overviewed. Presently, most stress management activities are oriented towards secondary or tertiary prevention, and are worker oriented. But in future, priority strategy for intervention should be primary prevention, and focused on the organization as the generator of risk. In the group of countries paying a lot of attention to work stress, health policies or legal framework at the national level and a variety of activities for stress prevention at the company level are well integrated. By analyzing various stress management cases or projects, key factors for a successful approach to stress prevention are extracted as follows: 1. A stepwise and systematic approach, 2. clear determination of aims, tasks, responsibilities, planning and financial means, 3. An adequate diagnosis of risk analysis, 4. A combination of work directed and worker-directed measures, 5. A participative approach, 6. Top management support. Costs-benefit assessment should be introduced to evaluate the effectiveness of stress prevention and to promote more integrated approaches in the workplace. PMID- 11889822 TI - [Modification of serum lipid concentrations in young men exposed to a hot environment]. PMID- 11889823 TI - [Changes in urea cycle enzymes in rat liver caused by inorganic mercury]. PMID- 11889824 TI - [A case study of stress management by means of a self rating stress survey and interviews with employees]. PMID- 11889825 TI - [Investigation of the cab of steam locomotive from the viewpoint of occupational health and prevention of the accident occurred in the cab--the trouble on Karikatsu tunnel]. PMID- 11889826 TI - [Relationship between cigarette smoking and oral health status]. AB - The purpose of the study was to examine the relationship between the smoking habit and oral health status, while adjusting for age and some aspects of dental health behavior. The data used were based on a cross-sectional study on dental checkup in the worksite, which included a self-reported questionnaire and oral examination by a dentist. The oral health status variables were CPITN scores, missing teeth/filled teeth/decayed teeth, and self-reported gum bleeding. In addition, the subjects reported in a questionnaire concerning their smoking habit and dental health behavior. Of a total of 7,713 of workers, 5,232 (67.8%) participated in the dental checkup of the worksite. From the population, only the data for 3,303 men were analyzed. We used multiple logistic regression to calculate the odds ratio (OR) for each oral health status according to the smoking habit. Current smokers, compared to subjects who had never smoked, had a higher risk of periodontal disease (OR = 2.3; 95% CI: 1.9-2.7), missing teeth (OR = 1.6; 95% CI: 1.3-1.9) and decayed teeth (OR = 1.5; 95% CI: 1.3-1.8), but they had a reduced risk of gum bleeding (OR = 0.7; 95% CI: 0.6-0.8). Dose response relationships between smoking and these variables were also observed. The results indicated that cigarette smoking was associated with oral health status independent of some aspects of dental health behavior. PMID- 11889827 TI - [Abnormal myocardial contractile regulation mechanism in familial hypertrophic and dilated cardiomyopathies--functional analysis of molecular mutant troponin in in-vitro]. PMID- 11889828 TI - [A study of the feasibility and efficacy of fetal surgical closure of spontaneous cleft lip in CL/fraser mice]. AB - Cleft lip with or without cleft palate is one of the most common human craniofacial congenital malformations. Failure to form a continuous tissue in the upper lip during the early embryonic period results in cleft lip. There has been an increasing interest in the possibility of in-utero correction of life threatening fetal abnormalities. In the present study, the feasibility and efficacy of surgical suturing of the fetal mouse lip was examined by using CL/Fraser mouse fetuses (spontaneous of cleft lip) at embryonic 15, 16 or 17 days. The obtained results were as follows. #1. Abortion rate of pregnant dam applied inhalation of Ethrane was less than that of intraperitoneal injection of Nembutal. #2. Successful delivery rate of fetal mouse surgically treated at the embryonic 16 day or 17 day is higher than that on the embryonic 15 day. #3. Fusion rate of mouse fetal lip surgically treated on the embryonic 16 day was higher after 60 hours in dam's uteri than that after 36 hours. These results suggest that inhalation of Ethrane is more recommendable, and that the optimal timing of fetal surgery of spontaneous cleft lip in CL/Fr. mouse fetus is an embryonic 16 day, and that the average time of 60 hours after surgical operation is enough for sutured lip to fuse in dam's uteri. PMID- 11889829 TI - [Why does smoking cause lung cancer?]. PMID- 11889830 TI - [Mechanisms involved in large subcortical infarcts]. AB - Large subcortical infarcts(maximum diameter of infarct > or = 20 mm) result from various stroke patterns, including striatocapsular infarcts (SCI), corona radiata infarcts, centrum semiovale infarcts, and internal borderzone infarcts. A systematic investigation of stroke pathogenesis involved in large subcortical infarcts, however, has not been performed. This study attempted to clarify the stroke mechanisms involved in large subcortical infarcts, by examining 50 patients with large subcortical infarcts out of 430 ischemic stroke patients consecutively registered in our department. The subjects were divided into two groups according to the vascular territories involved on the MRI: 1) the lenticulostriate arteries group for 39 patients whose infarcts were restricted to within the vicinity of the lenticulostriate arteries; 2) the internal borderzone group for 11 patients whose infarcts mainly involved the internal borderzone (the upper part of the corona radiata and the centrum semiovale) between the territories of the deep perforating branches from the basal cerebral arteries and the medullary branches from the superficial pial arteries. Stroke pathogenesis were classified into the following 6 categories: A) cardiogenic embolism, 9 patients; B) artery-to-artery embolism, 6 patients; C) cryptogenic embolism, 2 patients; D) thrombotic MCA (M1) occlusion, 9 patients; E) thrombotic ICA occlusion, 10 patients; F) undetermined cause, 14 patients. The lenticulostriate arteries group consisted of 9 patients with cardiogenic embolism, 6 with artery to-artery embolism, 2 with cryptogenic embolism, 8 with thrombotic M1 occlusion, and 14 with undetermined cause. The internal borderzone group consisted of 10 patients with thrombotic ICA occlusion and 1 patient with thrombotic M1 occlusion. The stroke pathogenesis of the undetermined cause is considered to be thrombotic occlusion at the orifice of the lateral lenticulostriate artery, a so called "branch atheromatous disease (BAD)". The patients in this group experienced a gradual onset, and did not have a cardiac source of the embolism or proximal large artery disease. Among the patients reported as having SCI, BAD may play a role in some cases, especially in those whose the cause was classified as "undetermined". In conclusion, the lenticulostriate arteries group exhibited a higher frequency of cerebral embolisms (cardiogenic embolism, artery-to-artery embolism, and cryptogenic embolism) and thrombotic M1 occlusion, whereas the internal borderzone group had a higher frequency of thrombotic ICA occlusion. PMID- 11889831 TI - [An autopsy case of dementia with motor neuron disease accompanying Alzheimer's disease lesion]. AB - We report the case of a 60-year-old man with autopsy-proven dementia with motor neuron disease (D-MND) and Alzheimer's disease lesion. The patient presented with clumsiness of his right hand at the age of 55 years old and subsequently developed dysarthria, weakness and atrophy of his upper limbs. He was unaffectionate towards his family, repeated the same phrase, and showed severe disorientation of time and place. Neurological examination on admission showed not only diffuse lower motor neuron signs, such as weakness, atrophy, fasciculation and areflexia in both upper limbs, but also dementia (HDS-R 9/30). He died of respiratory insufficiency. Neuropathological examination showed mild atrophy of the frontal and temporal lobes and anterior spinal roots. Microscopic examination of cortical sections revealed degenerative changes with simple atrophy and gliosis, and these changes were predominant in layers 1 and 2 of the frontal and temporal cortices. Using immunohistochemical staining, ubiquitin positive but tau-negative inclusions were frequently found in neurons of the hippocampal granular cell layers and temporal lobes. Many senile plaques and neurofibrillary tangles were present in all sections of the brain. Our final diagnosis was dementia with motor neuron disease accompanying Alzheimer's disease lesion, because of hypoperfusion in the parietal lobe as well as the frontal lobe demonstrated by SPECT, and the presence of many senile plaques and neurofibrillary tangles in the cerebral cortex. Overlapping of pathologically proven D-MND and Alzheimer's disease lesion is extremely rare, and this case may improve our understanding of the process of neurodegeneration. PMID- 11889832 TI - [A case of MELAS presenting complex partial status epilepticus]. AB - We reported a 37-year-old man who presented complex partial status epilepticus as the initial symptom of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). He showed fluctuating consciousness disturbance, left homonymous hemianopsia, and paroxysmal conjugated eye deviation to the left. The lactic acid level was elevated in blood and CSF, and ragged-red fibers were observed in the biopsied muscle. MRI demonstrated T2-prolonged lesions in the right occipito-parieto-temporal lobes. Since a mutation of mitochondrial DNA (A3243G) was identified, he was diagnosed as having MELAS. On an ictal record, high amplitude, rhythmic sharp waves were observed at right parieto-temporo occipital region. High amplitude slow waves were also observed on the right hemisphere, especially in the right frontal lobe. These ictal discharges gradually decreased at their amplitude and in frequency, and then ictal EEG turned to the interictal EEG. During an ictal period, conjugated eye deviation to the left side and consciousness loss were observed. These seizures were observed once every several minutes. During the interictal period, sharp waves and sharp wave complexes were observed frequently at right parietal and posterior temporal lobes. The venous injection of diazepam (10 mg) normalized EEG quickly. When consciousness loss, especially fluctuating, was observed in the patients of MELAS, complex partial status epilepticus should be considered. PMID- 11889833 TI - [A case of granulomatous angiitis of the central nervous system presented with subacute mental deterioration resembling diffuse white matter disease]. AB - We reported a 60-year-old man with granulomatous angiitis of the central nervous system (GACNS) manifesting as subacute mental deterioration. His first symptoms were nausea and vomiting which brought him to a hospital, where no abnormality was found except for gastritis. One month later, he began to feel dizziness and brain tumor was suspected by a neurosurgeon with the MRI findings such as abnormal T2 signal and swelling in his brainstem. While he was followed up, he gradually presented mental change, disorientation and dysmnesia with the abnormal T2 signal spreading over the cerebral white matter bilaterally. Corticosteroid therapy was started based on the suspicion of a lymphoproliferative disease, and his symptoms and the abnormal MRI findings improved. Then he was referred to our department for further evaluation. Because we could not find any evidence of systemic diseases and he had been almost fully recovered, we discontinued the therapy. Soon after that, his mental deterioration as well as the abnormal T2 signal lesions on MRI relapsed. By open brain biopsy, the diagnosis of GACNS was established, and steroid pulse therapy was started. His symptoms and the abnormal T2 signal lesions improved gradually and the steroid was tapered to the maintenance dose without remission. Since the laboratory and imaging findings are not specific for the diagnosis of the angiitis confined to the central nervous system, brain biopsy is recommended for these disorders. PMID- 11889834 TI - [A case of visual perseveration attack caused by transverse-sigmoid sinus dural arteriovenous fistula]. AB - A case of visual perseveration attack in the right superior quadrantic visual field caused by left transverse-sigmoid sinus dural arteriovenous fistula (AVF) was reported. A 75-year old right-handed man noticed that when he saw an object, the image sometimes persisted even after he looked away or the object was removed. The palinoptic images always appeared in the right superior quadrant visual field, consisted of a row of multiple objects all identical to the real one in shape, and were accompanied by photopsia. The palinoptic images disappeared within a few minutes. He was neurologically normal and showed no hemianopia or quadrantanopia on admission. EEG showed no epileptic discharge. CT scanning of the brain revealed a small high density area in the left occipital lobe. MRI demonstrated hyperintensity in the left inferior occipital lobe corresponding to the lower part of Brodmann's areas 18 and 19 on T2 weighted image and flow-voids caused by the dilated left occipital artery. 99mTc-ECD SPECT disclosed a decrease of regional cerebral blood flow in the left occipital lobe. Cerebral angiography revealed dural AVF fed by three branches of the left occipital artery and another branch of the left ascending pharyngeal artery with retrograde drainage into the left transverse sinus, sigmoid sinus, and dilated cortical veins. The left transverse and sigmoid sinuses were occluded. Visual perseveration disappeared following the treatment of the dural AVF by transarterial embolization. Because the lesions on MRI and SPECT improved after the treatment, these lesions were considered to represent not infarction but vasogenic edema due to venous congestion. We emphasized the role of the left inferior occipital lesion including the secondary visual cortex (Brodmann's areas 18 and 19) as the cause of visual perseveration in the right superior quadrantic visual field. PMID- 11889835 TI - [A patient of chronic graft-versus-host disease presenting simultaneously with polymyositis and myasthenia gravis]. AB - We report a patient of polymyositis and myasthenia gravis as manifestations of chronic graft-versus-hot disease (GVHD). A 48-year-old man was diagnosed as having chronic myelogenous leukemia at the age of 42 years, and had bone marrow transplantation (BMT) two years after the onset of the disease. Since he suffered from mild liver dysfunction and cutaneous involvement manifesting chronic GVHD, he was placed on prednisolone and cyclophosphamide. As his condition improved, the prednisolone was gradually tapered. Forty-one months after the BMT, the patient developed muscle pain and muscle weakness. A diagnosis of polymyositis was made from muscle biopsy and laboratory findings. An increase in the prednisolone dose was effective but a few weeks later the patient noticed ptosis and recurrence of muscle weakness. A tensilon test and anti-acetylcholine receptor antibody produced positive results, leading to a diagnosis of myasthenia gravis. Only one case of polymyositis and myasthenia gravis as manifestations of chronic GVHD has been reported, and in our patient both symptoms appeared almost at the same time. Although neuromuscular symptoms as a manifestation of chronic GVHD are rare, all patients receiving BMT should be carefully followed up neurologically to detect neuromuscular complications. PMID- 11889836 TI - [A case of cerebellar degeneration with schizophrenia-like psychosis, severe iron deficiency, hypoceruloplasminemia and abnormal electroretinography: a new syndrome?]. AB - A 33-year-old male patient began to develop schizophrenia-like symptoms and slowly progressive cerebellar ataxia. He was 170 cm tall and he had mild frontal baldness. Psychiatrically he was aconative, only willing to do nothing all day long after admission. He had neither hallucinations nor delusions, and his mental acuity was normal. Neurological examination revealed positive cerebellar signs including clumsiness in F-N-T and K-H-T and dysdiadochokinesis. He could neither stand up nor walk because of ataxia. The brain MRI showed severe cerebellar atrophy with normal basal ganglia. His EEG and the value of NCV were within normal range, whereas electroretinography showed a notable abnormality, pointing to the extremely small b-wave, resulting in a negative shape of the ERG. Although he was eating sufficiently, the level of serum iron and ferritin remained constantly low. The serum copper level was within normal range, whereas the serum ceruloplasmin level was mildly decreased. A hepatic biopsy indicated no accumulation of copper or iron. This case suggests the importance of the investigation of the serum iron and ceruloplasmin levels in patients who have cerebellar degeneration with psychosis. PMID- 11889837 TI - [A case of Machado-Joseph disease presenting pure cerebellar ataxia]. AB - We report a 61-year-old woman with Machado-Joseph disease (MJD) presenting with pure cerebellar ataxia. The patient exhibited an unsteady gait at the age of 51 years. She was admitted to our hospital at the age of 61 years. Her older brother had been diagnosed as having spinocerebellar degeneration (SCD). Our patient showed gaze-evoked nystagmus, wide-based gait, slight lack of coordination of the four extremities, mildly ataxic speech and slight decrease in the bilateral Achilles tendon reflexes. Babinski's sign was absent. Sensory impairments were not present and muscle tone and muscle strength were normal. There was no autonomic dysfunctions. MRI revealed moderate atrophy of the cerebellum and pons. We performed gene analysis of SCD using white blood cells from the patient, and the analysis showed 70 CAG repeats in the MJD1 gene, which is an abnormally high number of repeats. Compared with three reported cases of MJD presenting pure cerebellar ataxia, only our patient showed a nasal voice. The number of CAG repeats in the MJD1 gene of our patients was the most prolonged of the four cases. MJD should be considered in patients with familial SCD even if their neurological signs and symptoms outside the cerebellum are not obvious. PMID- 11889838 TI - [Quantitative evaluation of the effect of 3,4-diaminopyridine in a patient with Lambert-Eaton myasthenic syndrome using dynamic dynamometry]. AB - We reported a 72-year-old woman with Lambert-Eaton myasthenic syndrome. The chief complaint was weakness and atrophy of the thigh muscles, which prevented her from climbing stairs even with a handrail. Sensory and autonomic function was normal without amblygeustia. There was no malignancy found, and her serum anti-V/Q type voltage-gated calcium channel antibody was negative. Administration of 3,4 diaminopyridine (DAP), known to accelerate acetylcholine release, was very effective and she became able to climb stairs without a handrail. For evaluation of the therapeutic effect of DAP, the initial compound muscle action potential (ICMAP) on evoked electromyogram has been recommended because it provides highly sensitive and reproducible results. Unfortunately this method is usually applied to several particular distal muscles for technical reasons. In the present case, evaluation of the quadriceps femoris muscle was most important because it was most responsible for her disability. We attempted to measure the angular velocity and the angular acceleration on knee extension movement using dynamic dynamometry. The angular velocity improved from 124 to 162 deg/sec and the angular acceleration from 220 to 390 deg/sec2. The results were more sensitive and more relevant to her demonstrable ADL improvement than grasping power increase and ICMAP improvement recorded at the distal muscles. PMID- 11889839 TI - [Acute onset of tuberculous meningoencephalitis presenting with symmetric linear lesions in the bilateral thalamus: a case report]. AB - A 18-year-old woman was admitted to our hospital because of high fever and headache. Nuchal stiffness was present, and a CSF examination showed lymphocyte domonant pleocytosis and a decreased level of glucose. Although antibiotics, aciclovir and an antimycotic drug were administered, disturbance of consciousness, involuntary movements, and pyramidal tract signs appeared. Soon after the medications were changed to antituberculous medicines, the meningoencephalitis started to subside, and was finally cured. Judging from the clinical findings, the CSF findings, the effectiveness of antituberculous medicines, an elevated ADA level in CSF, and positive conversion in tuberculin tests, the final diagnosis was made as tuberculous meningoencephalitis. At the severest stage of the disease, a brain MRI showed symmetric, linear lesions without the effect of Gd-enhancement in the bilateral thalamus, which thereafter disappeared along with the healing of the illness. From all these things, we conclude that thalamic and other parenchymal lesions should be kept in mind in case of acute tuberculous meningoencephalitis. PMID- 11889840 TI - [Ataxic form of Guillain-Barre syndrome associated with anti-GD1b IgG antibody]. AB - A 28-year-old man was admitted after developing acute onset unstable gait following acute enteritis. Neurological examination revealed mild weakness in four limbs, areflexia and ataxia. Serum obtained from the patient during the acute stage contained a high titer of anti-GD1b IgG antibody. Because the patient showed obvious cerebellar ataxia unrelated to muscle weakness, without ophthalmoplegia or proprioceptive sensory disturbance, we concluded that he had ataxic form of Guillain-Barre syndrome (GBS) (Richter, 1962). Although ataxic GBS is not an established conception, one should pay attention to the possible existence of such a rare GBS variant. It is necessary to accumulate additional case reports to clarify the association between ataxic GBS and anti-ganglioside antibodies. PMID- 11889841 TI - [Animal-derived feedstuffs as possible vectors for bovine encephalopathy (BSE) in Germany. Part 2: Assessment of vector risk for compounded feed]. AB - Specific conditions and practices of cattle feeding in Germany have to be taken into account for assessing the risk of feed born transmission of bovine spongiform encephalopathy, especially regarding the situation before the year 2000 when specific directives were introduced for feed production. The present retrospective epidemiological study includes data on feed production and the estimated amount of animal derived feedstuffs for the production of compounded feed for cattle. Risk assessment was performed based on the 'reproduction rate' (R0), that is defined as the estimated number of infections resulting from the processing of brain and spinal cord of BSE affected cattle that is recycled to bovines via feed. Under the conditions as given in Germany until the year 2000 the reproduction rate of BSE via the inclusion of animal derived feedstuffs in compounded feed production for cattle was estimated to be 1.1. Thus, it can be expected that BSE could be reproduced in the system, but with comparatively low efficiency. The expected incidence of BSE should be considerably lower compared to the situation during the 90th in the UK, due to the markedly lower recycling rate of animal protein in cattle feeding. Animal fat could have been a significant factor for BSE transmission due to contamination by proteinaceous brain and spinal cord material during the production process. The relative significance of fat containing feedstuffs for BSE transmission could have been higher in Germany compared to the situation in the UK where meat and bone meal was produced under different conditions and frequently used in higher proportions as an ingredient for compounded feed for ruminants. PMID- 11889842 TI - [Suitability of sonographic evaluation of ovarian dynamics and uterine involution for prediction of postpartum fertility in the cow]. AB - To investigate the suitability of sonography for prediction of puerperal fertility, we used 34 cows with normal puerperium. For this purpose, the animals were examined gynecologically and by transrectal sonography at 10-day intervals between day 15 and day 45 p.p. as well as at oestrus, when they were inseminated. We judged uterine involution by diameter and sonographic structure of the uterine cross-section and ovarian activity by the presence of follicles and corpora lutea. Cows, which became pregnant after first insemination p.p., were assigned to group G1 (n = 16) and animals, which failed to conceive, to group G2 (n = 18). The cows of G1 obtained significantly better results concerning both uterine involution and ovarian activity than cows of G2. Thus, diameter of the uterine horn decreased faster and sonographic structure of the uterine cross section returned markedly earlier to the nonpregnant appearance in G1 than in G2. Group G1 also showed a significantly higher percentage of dominant follicles at all examinations. The results of the study show a distinct relationship between uterine involution and ovarian activity and puerperal fertility. But it has also to be resumed that the prediction of fertility for individuals will always be restricted by other reasons causing failure of conception. PMID- 11889843 TI - [Fetotomy in cattle with special reference to postoperative complications--an evaluation of 131 cases]. AB - In the course of this retrospective study on fetotomy of cattle 131 operations were evaluated. The case studies put the cattle 2 to 96 hours into labour (average 13 hours). The cows that exhibited post-operative complications were on average 19.2 hours into labour prior to the operation. Those which had a normal puerperium however were only 8.4 hours, on average, in labour. 79% of the animals showed depressed general health on admission, 8% of which were unable to stand. The main cause of the dystocia was the incorrect position/orientation of the dead foetus (38.9%) as well as relative or absolutely too large calves (25.2%). In 14.5% of the cases the offspring were malformed causing the dystocia. 28 of the protracted births were due to both maternal as well as foetal causes. Mortality of the cows lay by 6.9%, irregularities in the puerperal period ran at 67.2%. The age of the mother did not seem to influence the post operative events. The most common complication was retained placenta (n = 49; 37.4%) followed by lochiometra (n = 21; 16%), vaginal wounds and pelvic phlegmons (each n = 16; 12.2%) and neurotripsy (n = 6; 12.2%). Of these puerperal complications the pelvic phlegmon required the longest post-operative care (14.3 days, retention secundinarum was cured in 13 days, lochiometra in 8.8, cows without complaint were treated for 3.8 days after the operation). The most important factors for the post-operative prognosis are the time in labour and any trauma of the soft birth canal which influence the forming of pelvic phlegmons. PMID- 11889844 TI - [Successful direct transfer of a deep frozen-thawed equine embryo]. AB - Embryos were flushed on day 7 after ovulation from two mares, and frozen using a conventional slow freezing procedure in phosphate buffered (PBS) saline supplemented with 10% FCS, 1.5 mol/L ethylene glycol and 0.25 mol/L sucrose. One of the two embryos was thawed after 10 months of storage in liquid nitrogen and transferred directly (without dilution of the cryoprotectant and quality examination) to a synchronized recipient. This transfer resulted in the birth of a live female foal. To our knowledge, this is the first live foal born after direct transfer of a frozen-thawed equine embryo. PMID- 11889845 TI - [The dog welfare directive of 2 May 2001]. AB - As of 1 September, 2001 existing legislation regulating outdoor dog husbandry is replaced by the domestic dog welfare directive of 2 May, 2001. The new directive applies to details of housing and breeding of dogs kept indoors and as domestic companions. Thus, minimum requirements for housing, care and feeding now apply to the great majority of private dog owners. However these requirements do not apply to animals in transit, in individual veterinary treatment or to those used for scientific experimentation whose goals justify deviations from these standards. Additional regulations apply to dog breeding and showing. Puppies may not be separated from their mother and litter before eight weeks of age. "Prohibition of breeding for aggressiveness" cited in Paragraph 11b of the animal welfare law is defined, and includes various breeds and hybrids. The directive includes an index for the care and treatment of dogs by commercial breeders. After a transitional period no dogs which have been subjected to amputations at the expense of the animal's welfare in order to achieve specific features may be shown publicly. The author provides critical commentary on the regulations of the domestic dog welfare directive and points out areas which will continue to be of concern. PMID- 11889846 TI - [Statistical principles of "good clinical practice" in veterinary medicine--a position paper for planning, implementation and evaluation of empirical studies]. AB - The complexity of the design, conduct, analysis and evaluation of empirical studies necessitates a high degree of interdisciplinary collaboration in all areas of research. In order to make sure that no essential harmonisation is missed among the plenitude of processes, it has become common to provide direction on the essential operating procedures in so called "Good ... Practice" guidelines in recent years. In pharmaceutical research on human medicinal products guidelines on Good Clinical Practice have been an integral part of research and development in industry, academia, and the regulatory authorities for a long time. On the other hand, in the development and registration of pharmaceuticals for veterinary use such procedures are not yet established to this extent in Germany. Notwithstanding there being a lot of regulations on specialised subjects. This paper tries to summarise the current state of the discussion and to give an overview on the important points in the design, conduct, analysis and reporting of veterinary clinical studies mainly from an biometrical point of view. PMID- 11889848 TI - Activities of epidemiologists in state and territorial cancer control programs, 1999. AB - The Council of State and Territorial Epidemiologists surveyed the chronic disease program directors in 1999. Forty-eight (89%) of the health agencies responded. Nearly 90 percent reported having an epidemiologist who devoted some time to cancer control activities. More than 90 percent reported the availability and use of the mortality, incidence, and risk factor data. Almost 70 percent of the respondents produced annual cancer reports, but less than half reported publishing in a state bulletin or scientific journal. These findings suggest that despite widespread access to epidemiologic expertise and cancer-related data, considerable variability persists in the use and dissemination of crucial cancer statistics. PMID- 11889847 TI - Colorectal cancer screening among individuals with and without a family history. AB - Data from a community-based screening campaign were analyzed to assess compliance with colorectal cancer (CRC) screening guidelines among both average risk adults and those at increased risk because of having a first-degree relative (FDR) with CRC (FDR+). The prevalence of screening compliance was low in both groups. The authors also found that individuals with FDR+ returned a free fecal occult blood test at a slightly higher rate than those without FDR+s. Despite higher screening rates among individuals with FDR+s, interventions may need to target this high risk group to increase compliance with CRC screening recommendations. PMID- 11889849 TI - Use of a geographic information system to identify and characterize areas with high proportions of distant stage breast cancer. AB - A spatial scan statistic was used to search for geographic areas with significantly elevated proportions of women diagnosed with distant stage breast cancer in New Jersey in 1995-1997. The identified areas then were mapped and characterized using data from the 1990 U.S. Census and locations of mammography facilities. These areas' population characteristics included relatively high proportions of black or Hispanic women and linguistically isolated households. Targeted education and screening programs using this information may increase the diagnosis of breast cancer in the early stages, thereby reducing breast cancer mortality. PMID- 11889850 TI - Small area analysis on a large scale--the California experience in mapping teenage birth "hot spots" for resource allocation. AB - Small-area analysis has become an important tool in the effective targeting of limited public health resources. In California, new funding for teenage pregnancy prevention programs required more and better information to justify the allocation of these funds to areas with the greatest need. Consequently, these funds were allocated using maps with census tract analyses of teenage birth rates and an overlay of geographic frequencies. State and local agencies' programs have responded with positive feedback to the maps, and public health management subsequently has augmented funding for mapping equipment and training. The lessons learned and future directions are discussed. PMID- 11889851 TI - Hepatitis C prevention programs: assessment of local health department capacity. AB - Hepatitis C virus (HCV) infection is the most common bloodborne infection in the United States. To determine the capacity of local health departments to respond to concerns about HCV, local health officers were surveyed regarding HCV programs and needs. Of 612 respondents, fewer reported offering HCV services (education, counseling, testing) compared with those for HIV. Most respondents reported that technical assistance would be needed for HCV services and that such services should be integrated into existing HIV programs. Many local health departments may be unprepared for a growing need for public HCV services; integrated HCV-HIV programs should be considered. PMID- 11889852 TI - The forms that bind: multiple data forms result in internal disaggregation of immunization information. AB - To examine how forms encountered during routine clinical activities impact a provider's immunization activity, workflow analysis was performed in nine community clinics and small private practices. Data gathered included the number, source, and nature of forms. A total of 200 forms were used by the nine clinics just for children under 35 months of age. These represent a real labor cost as well as an opportunity cost. Use of a single summary sheet, yearly review of the forms, and coordination of agency documentation efforts are recommended. PMID- 11889853 TI - Linking WIC and immunization services to improve preventive health care among low income children in WIC. AB - Children participating in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) are at risk for low immunization coverage and other adverse health-related outcomes. Immunization-promoting strategies in WIC have been shown to produce dramatic improvements in immunization coverage. This evaluation of a local WIC initiative in Milwaukee is the first study to evaluate the impact of these strategies on improving the utilization of other clinical preventive services at the medical home. The use of more intensive immunization promoting strategies in WIC may improve utilization of well child care visits and receipt of other clinical preventive services in the medical home. PMID- 11889854 TI - The Management Academy for Public Health: a new paradigm for public health management development. AB - Research has established a need to develop management skills among public health professionals. The University of North Carolina-Chapel Hill created the Management Academy for Public Health as a pilot program for this specialized training need. This article describes why a management academy for public health managers was formed, its curriculum and instructional methods, and the evaluation findings from its first year. The program sponsors hope to effect individual and organization level change, eventually leading to improved community health. Results suggest that this innovative program gives public health professionals needed skills and improves their job performance. PMID- 11889855 TI - Organizational and community health changes under welfare reform. AB - This article comments on changes made by health-related organizations and in community health during the first years of New Jersey's welfare reform policy implementation in Camden, Essex, and Hudson counties (1992-1998). Throughout these years, county welfare agencies made organizational changes that shifted their focus from supporting family well-being to encouraging work. Health care provider organizations now are beginning to make organizational changes in response to the ripple effects of welfare reform policy. Specific actions to initiate and influence healthy welfare policy reformulation are presented. PMID- 11889857 TI - Overtime--mandatory or voluntary? PMID- 11889856 TI - Public health workforce development: keeping population health goals in mind. AB - In the public health field, there is an identified need to develop the public health workforce strategically to meet the needs of a changing public health landscape. The "core competencies" that support the implementation of the core functions of public health must be tied to the mission and goals of the agency or program, and examined in light of the specific population health concerns they are meant to address. CAST-5 is offered as an example of a tool fulfilling this role for state Maternal and Child Health programs. PMID- 11889858 TI - Scrub nurse practice. An ethical viewpoint. PMID- 11889859 TI - Local anaesthetic techniques in ophthalmic surgery. AB - This article, which won third place in the Alison Bell Writer's Award this year, reports the findings of a literature review that explored the use of local anaesthetic techniques used in ophthalmic surgery. The author describes the various ophthalmic procedures that can take place under local anaesthetic and asks whether there is a need for an anaesthetist to be present for such lists. Issues such as patient monitoring requirements and intravenous access are also discussed, emphasising the importance of the nurse's role in informing, supporting and comforting the patient through what can be a stressful experience. PMID- 11889860 TI - [Pressing problems of orthodontics in the journal "Stomatologiia" over 80 years]. PMID- 11889861 TI - [Topics of publications in the journal "Stomatologiia" during the recent decade]. PMID- 11889862 TI - [Morphogenesis of early stages of periodontal inflammations]. AB - Morphological signs of dissemination of active inflammatory process beyond the gingiva into the depth of the alveolar process bone tissue, paralleled by numerous destructive changes (bone resorption and lysis of collagen fibers of the periodontal ligament, plunged into the bone) in the periodontium are observed during clinically manifest chronic gingivitis. Hence, in contrast to the concept universally acknowledged in periodontology, despite the seeming clinical "heterogeneity" of gingivitis and periodontitis, these conditions should be regarded as a periodontal inflammation, because, judging by the detected morphological picture, each of the nosological entities acknowledged today (chronic gingivitis, periodontitis) represents just a successive stage of the same chronic inflammatory process differing only quantitatively but not qualitatively. PMID- 11889863 TI - [Acoustic microscopy methods in studies of the filling materials microstructure]. AB - The structure of filling materials (kemfil, polyacrylate cement, and Dentis, material produced by Stomadent Firm in Russia) at the interface with human dentin was studied in teeth subjected to retrograde filling with preliminary appendectomy. A prototype of table wide-field short pulse scanning acoustic microscope (50 mHz) (Acoustic Microscopy Center, Institute of Biochemical Physics of Russian Academy of Sciences) and Elsam acoustic microscope (200 mHz) (Leitz, Germany) were used. The results indicate that analysis of acoustic characteristics, including the data of acoustic microscopy, helps investigate the cement microstructure and evaluate the compactness and elasticity of samples; moreover, structural elements of the material, which are undetectable by other methods, are seen on acoustic images. These data can be used for evaluating the relationship between the microstructure and formation of mechanical properties, and, maybe, the patterns of cement interactions with dental tissues. PMID- 11889864 TI - [On the mechanism of tactile sensitivity of teeth]. PMID- 11889865 TI - [Composite materials of different classes in practical dental surgery]. AB - Presents the indications for the use of autohardening composite materials in dentistry and validates the need in introduction of materials of different classes along with universal hybrid composites. Sums up the results of introduction of these materials into practice and discusses the trends in the practical use of photohardened composites in Russia. PMID- 11889866 TI - [Use of probiotics Bifidumbacterin and Acilact in tablets in therapy of periodontal inflammations]. AB - The results of the use of russian probiotics Acilact and Bifidumbacterin in complex treatment of gingivitis and different degrees of parodontitis are presented. The treatment of the patients of control group was added by the italien drug Tantum Verde. The effect of Acilact and Bifidumbacterin to the normalisation of Mycroflora higher in comparison with Tantum-Verde, particulary in the cases of gingivitis and parodontitis superficialis. PMID- 11889868 TI - [Laser fluorescent diagnostic methods in purulent surgery]. PMID- 11889867 TI - [Validation of the optimal concentration of Metrogil-denta preparation in the treatment of periodontal inflammations]. AB - Fifty-two patients (22-68 years) with chronic generalized periodontitis of different severity and 223 with fulminant severe periodontitis were observed. The bacteriostatic and bactericidal effects of Metrogil-denta depended on the severity of periodontal involvement and metronidasole concentration in the drug. The maximum bacteriostatic effect was observed at 20-25% and the maximum bactericidal effect at 25% concentration of metronidasole in the preparation irrespective of periodontitis severity. PMID- 11889869 TI - [Leader of Russian research and practical dentistry ]. PMID- 11889870 TI - [Results and perspectives of development in dental implantation in Russia]. AB - The basic trends of scientific research carried out at the implantation department of CRID (Central Research Institute of Dentistry), referred to the fundamental problems in medicine and biology including a variety of not only experiments and clinical study but also a complex of technical aspects. Our long term research and valuable clinical material collected resulted in confirming the necessity and importance of complex approaches developed by the Institute in dental implantation. PMID- 11889871 TI - [Mathematical simulation of mechanical system abutment teeth--aramide thread adhesive bridge denture]. AB - A mathematical model of a new pontic design is developed. The final element analysis of the model proved the efficiency of the design. Effects on the denture length, abutment area cone angle, thread strain and height of location on abutment teeth, depending on the type and site of exercise, were studied. PMID- 11889872 TI - [Fluoride metabolism in children and caries prevention]. AB - Basal mineral elements content in superficial dental enamel layers and daily urine fluoride excretion have been investigated in children using water with various level of this trace element. The investigation was carried out before and after sodium fluoride intake in the doses 1.1-1.6 mg/day. The results give evidence that differential approach to dental caries prevention with fluoride is required. PMID- 11889873 TI - [Calcium homeostasis disorders in children with multiple caries]. AB - A saliva of the examined children of 3, 6, 12 years old with intensive caries are characterized by a changing of calcium homeostasis which is accompanied with redistribution of calcium fractions in a mixed nonstimulation saliva. In this case the concentration of Ca2+ is raisign considerably while the concentration of general calcium is raising slightly. PMID- 11889874 TI - [Theoretical problems in the journal "Stomatologiia"]. PMID- 11889875 TI - [Correction of central mechanisms of sanogenesis regulation in children with cleft palate]. AB - Theoretical prerequisites for the development of new methods for treatment of children with cleft palate by physical factor exposure are presented. Morphofunctional characteristics of the central nervous system formation during the prenatal and early postnatal periods are described. A multi-level method of magnetic laser exposure of the cerebrocortical precentral gyrus and electrostimulation by the ciliary reflex system, modifying the central regulation and adaptation mechanisms during the immediate period after uranoplasty, has been developed. PMID- 11889876 TI - [Dentistry in Russia: state of the art and prospects for development]. PMID- 11889877 TI - Capitation: the wrong direction for primary care reform. PMID- 11889878 TI - Argument for blended funding. PMID- 11889879 TI - Canadian Family Physician's new ethics guidelines. PMID- 11889880 TI - Capitation by any other name.... PMID- 11889881 TI - Academic family medicine a lot like yeast. PMID- 11889882 TI - Symbols and spellings in our December issue. PMID- 11889883 TI - "Help me breathe." Finding the right way to respond. PMID- 11889884 TI - Nausea and vomiting of pregnancy. Evidence-based treatment algorithm. AB - QUESTION: One of my patients suffers from a moderate-to-severe form of morning sickness. She responded only partially to doxylamine and pyridoxine (Dicletin), and I wish to try adding another medication. What should my priority be? ANSWER: An algorithm used by Motherisk to manage thousands of patients takes a hierarchical approach to this condition. This approach is evidence based with regard to fetal safety as well as efficacy. PMID- 11889885 TI - Dermacase. Erosio interdigitalis blastomycetica. PMID- 11889886 TI - Practice tips. Treating persistent cough. Try a nebulized mixture of lidocaine and bupivacaine. PMID- 11889887 TI - Can we prevent high-risk patients from getting type 2 diabetes? PMID- 11889888 TI - Corticosteroid injections and arthrocentesis. AB - OBJECTIVE: To review current standards of practice of arthrocentesis and corticosteroid injections in soft tissue and joints in managing common musculoskeletal conditions. To outline common indications, contraindications, and possible complications of these therapeutic modalities and to describe common techniques used in them. QUALITY OF EVIDENCE: Many of our recommendations are based on expert opinion and surveys of clinical practice by experts in the field. Where appropriate, randomized controlled trials are used to support various aspects of corticosteroid injection and joint aspiration. MAIN MESSAGE: Many complaints to primary care physicians are musculoskeletal in origin, and many are related to infectious or inflammatory conditions of joints or soft tissues. Arthrocentesis and intra-articular and soft tissue corticosteroid injection are therapeutic techniques readily available to family practitioners. There is no consensus on best practice for patient preparation, choice of corticosteroid, or specific injection technique. There are, however, accepted standards of practice and universal precautions. CONCLUSION: Family physicians have an important role in managing many common musculoskeletal conditions. PMID- 11889889 TI - Indoor air quality, fungi, and health. How do we stand? AB - OBJECTIVE: To equip medical practitioners with up-to-date scientific and medical information on the health effects of exposure to fungi in indoor air, clinical evaluation of these health problems, and possible preventive measures. QUALITY OF EVIDENCE: MEDLINE was searched from 1985 to 2000 using the MeSH words mould (mold), fungus, indoor air, and health effects. Nearly all studies were case reports, case-control studies, and cross-sectional studies. Evidence of an association between respiratory problems and the presence of fungi and dampness is strong. MAIN MESSAGE: Recent well designed studies and literature reviews indicate that exposure to dampness and fungi in indoor air brings on or exacerbates asthma and other respiratory complaints. More studies are required, however, before a definite conclusion on other potential effects of such exposure (such as systemic and long-term effects and pulmonary hemorrhage in infants) is possible. The various health problems that can result from exposure to dampness and fungi in indoor air make such exposure unacceptable from a public health perspective. Physicians are important in treating and preventing such problems; various resources are available to help them. CONCLUSION: Even though some scientific issues remain to be resolved concerning the health effects of exposure to dampness and fungi in indoor air, family physicians can identify potential problems and refer patients to appropriate resources. PMID- 11889890 TI - Primary care reform. Physicians' participation in Hamilton-Wentworth. AB - OBJECTIVE: To determine physicians' reasons for and against participating in a primary care reform (PCR) pilot project, to identify demographic and practice characteristics associated with participation, to gauge physicians' satisfaction with implementation of the project, and to seek suggestions for change. DESIGN: Cross-sectional mailed survey using a self-administered questionnaire. SETTING: Family practices in Hamilton-Wentworth, Ont. PARTICIPANTS: Eighty-two of 107 (76.6%) physicians who participated in the pilot project and 101 of 150 (67.3% of a 60% random sample of the area's remaining generalist physicians) who chose not to participate. MAIN OUTCOME MEASURES: Physicians' primary and secondary motives for participating or not; comments on the pilot project; and subjects' demographic, professional, and practice characteristics. RESULTS: Despite their experience with capitation practice, after controlling for other factors, physicians from health service organizations were no more likely than their fee for-service colleagues to join the pilot project. Those in large group practices were more likely to participate. Both participants and non-participants were concerned about disrupting call groups, burdening office staff, not having enough time, and whether the project's objectives were achievable. Other key findings were how few patients declined enrolment and how many physicians had unrealistic ideas about the demands of participation and the capabilities of currently available information technology. CONCLUSION: While many Hamilton-area physicians were eligible and willing to participate in a PCR pilot project, many were not. Our findings suggest that physicians and government should clarify their expectations for PCR and that we need to look for better ways to register patients and select information technology for PCR. PMID- 11889891 TI - Peril of the pox. Are primary care providers aware of varicella vaccination guidelines? AB - PURPOSE: To determine whether physicians providing primary care in Ontario were aware of guidelines published by Health Canada's National Advisory Committee on Immunization (NACI), their opinions about the varicella vaccine, and factors that influence use of the vaccine. METHODS: A questionnaire examining awareness, knowledge, and perceived barriers to using varicella vaccine was developed and mailed to a random sample of 500 family physicians and 400 pediatricians practising in Ontario. RESULTS: To the 900 questionnaires mailed, 284 family physicians (56.9%) and 232 pediatricians (59.2%) responded. Fifty-six percent of family physicians and 95.4% of pediatricians providing primary care were aware of the Health Canada guidelines. Physicians who were aware of the Health Canada guidelines were more knowledgeable about the vaccine and were more likely to recommend it. Both groups of physicians identified cost (not covered by the government), problems with storage and handling, and concerns about long-term immunity as barriers to use of the vaccine. CONCLUSION: While awareness of the guidelines was associated with better knowledge of and following recommendations for the vaccine, Health Canada guidelines were not widely distributed to all primary care providers. One way to encourage incorporation of varicella vaccine guidelines into practice would be to improve dissemination of the guidelines to all primary care providers. PMID- 11889892 TI - Anesthesia skills for rural family physicians. PMID- 11889894 TI - On the forum and the future. PMID- 11889893 TI - Medical websites for users of hand-held computers. PMID- 11889895 TI - Exploiting antibodies as catalysts: potential therapeutic applications. AB - In this review, we explore recent developments in the generation of catalytic antibodies and their potential in therapy. PMID- 11889896 TI - Recombinant monoclonal antibody technology. AB - With the development of murine hybridoma technology over a quarter century ago, the ability to produce large quantities of well-characterized monoclonal antibody preparations revolutionized diagnostic and therapeutic medicine. For many applications in transfusion medicine, however, the production of serological reagents in mice has certain biological limitations relating to the difficulty in obtaining murine monoclonal antibodies specific for many human blood group antigens. Furthermore, for therapeutic purposes, the efficacy of murine-derived immunoglobulin preparations is limited by the induction of anti-mouse immune responses. Technical difficulties inherent in human hybridoma formation have led to novel molecular approaches that facilitate the isolation and production of human antibodies without the need for B-cell transformation, tissue culture, or even immunized individuals. These technologies, referred to as 'repertoire cloning' or 'Fab/phage display', involve the rapid cloning of immunoglobulin gene segments to create immune libraries from which antibodies with desired specificities can be selected. The use of such recombinant methods in transfusion medicine is anticipated to play an important role in the development and production of renewable supplies of low-cost reagents for diagnostic and therapeutic applications. PMID- 11889897 TI - Section 1A: Rh serology. Coordinator's report. AB - One hundred forty-two Rh-specific monoclonal antibodies (Mabs) were evaluated by serology in 27 laboratories. Evaluators were asked to test each Mab at three dilutions in specified serological techniques against normal positive and normal negative phenotype cells, and any Rh variant cells that they had available. Raw data was submitted to the coordinator for overall analysis. Results were analysed by expressing the sum of reaction grades for each Mab with each variant cell as a percentage of the sum of reaction grades of that Mab with normal phenotype cells. Anti-D Mabs were sorted into 23 groups which had the same pattern of reactions with different partial D phenotype cells. Eighteen of these corresponded to previously defined patterns; five were new patterns. Combined with data from the previous workshop, this means that 30 different reaction patterns have been defined. A new nomenclature is introduced for numbering the epitopes. Reactions with new variants DNB, DNU and DAR indicated some further subsplits of these patterns. Reactions with Category Va cells indicated that there were five different types of Va cells that could be distinguished serologically with monoclonal antibodies. No patterns of reactivity corresponding to the epitope groups could be observed with the different types of weak D tested. Anti-E Mabs were sorted into 14 groups, and the E variant cells into seven groups. PMID- 11889898 TI - Section 1B: Rh flow cytometry. Coordinator's report. Rhesus index and antigen density: an analysis of the reproducibility of flow cytometric determination. AB - Fifty-seven IgG monoclonal anti-D antibodies were evaluated in the Rh flow cytometry section, in which 12 laboratories participated. Staining protocols and a fluorescein (FITC)-conjugated Fab fragment goat anti-human IgG (H + L) as a secondary antibody were recommended but not mandatory. A CcDEe red blood cell (RBC) sample that was shown to be homozygous for RHD by molecular methods was supplied and used as internal 'standard RBC' throughout all experiments. An RBC panel comprising two partial D and four weak D types was supplied as well. The use of standard RBC reduced the variability of the data among the laboratories and allowed the conversion of fluorescence data into epitope densities, which were compounded in an antigen density (antigen D per RBC). The highest antigen density was determined for DVI type III, followed by DVII and weak D type 3; the lowest antigen density were determined for weak D type 1 and type 2. Nine of the 12 participating laboratories discriminated three groups of aberrant RhD that had similar Rhesus indices (RI): D category VI with RI = 0; weak D type 2 and type 3 with an high RI; and D category VII and weak D type 1 with an intermediate RI. The antigen densities and the Rhesus indices obtained correlated well among the laboratories of this Workshop section despite different staining protocols, secondary antibodies and instrumentation. PMID- 11889899 TI - Section 1C: Assessment of the functional activity and IgG Fc receptor utilisation of 64 IgG Rh monoclonal antibodies. Coordinator's report. AB - Sixty-four IgG Rh monoclonal antibodies (Mabs) submitted to the Fourth International Workshop on Monoclonal Antibodies Against Human Red Blood Cells and Related Antigens were characterised and tested in quantitative functional assays at five laboratories. The biological assays measured the ability of anti-D to mediate phagocytosis or extracellular lysis of RBC by IgG Fc receptor (Fc gamma R)-bearing effector cells. Interactions of RBC pre-sensitised with anti-D (EA IgG) with monocytes in chemiluminescence (CL) assays were found proportional to the amount of IgG anti-D on the RBC. Using antibodies to inhibit Fc gamma RI, Fc gamma RII or Fc gamma RIII, the only receptor utilised in the monocyte CL and ADCC assays for interactions with EA-IgG1 was found to be Fc gamma RI. In these assays, enhanced interactions were promoted by EA-IgG3 and additional Fc gamma receptors may have contributed. IgG2 anti-D was not reactive in these assays and EA-IgG4 promoted weak reactions through Fc gamma RI. A macrophage ADCC assay showed that haemolysis of EA-IgG3 was greater than that of EA-IgG1, mediated mainly through Fc gamma RIII. In ADCC assays using lymphocytes (NK cells) as effector cells and papainised RBC target cells, only a minority of IgG1 anti-D Mabs were shown to be able to mediate haemolysis in the presence of monomeric IgG (AB serum or IVIg). These interactions were mediated solely through Fc gamma RIII. Haemolysis via Fc gamma RIII may depend on the presence of certain sugars on the oligosaccharide moiety of IgG. Most Mabs (IgG1, IgG2, IgG3 and IgG4) elicited intermediate, low or no haemolysis in these assays. Blocking studies indicated that low activity IgG1 and IgG4 anti-D utilised only Fc gamma RI. Other IgG1 and IgG3 Mabs appeared to promote haemolysis through Fc gamma RI and Fc gamma RIII while IgG2 was inhibited by Mabs to both Fc gamma RII and Fc gamma RIII, suggesting a variety of Fc gamma R are utilised for anti-D of low haemolytic activity. Excellent agreement between the results of the lymphocyte ADCC assays and antibody quantitation was observed between the participating laboratories. PMID- 11889900 TI - Section 2: Immunochemical, immunohistological and serological analysis of monoclonal antibodies with carbohydrates. Coordinator's report. PMID- 11889901 TI - Section 3: Epitope determination of monoclonal antibodies to glycophorin A and glycophorin B. Coordinator's report. Antibodies to antigens located on glycophorins and band 3. PMID- 11889902 TI - Section 4: Antibodies to other blood group antigens. Coordinator's report. PMID- 11889903 TI - Section 5: Structural/genetic analysis of mAbs to blood group antigens. Coordinator's report. AB - The heavy and light chain immunoglobulin variable region nucleotide sequences for 219 mAbs to human red blood cells were collected from workshop participants, published reports, and Genbank. Information regarding antigen specificity, species of origin, method of cloning, and other relevant serological properties was correlated with the sequence data. Immunoglobulin sequences were analyzed to determine the heavy- and light-chain immunoglobulin genes used and the overall extent of somatic mutation from germline configuration. Approximately 50% of the sequences encoded antibodies with Rh(D) specificity with the remaining sequences encoding mAbs to other Rh-related antigens, antigens of the ABO, MNS, and Kell blood group systems, and several others. Surprisingly, no sequence data were available for mAbs with specificity for a number of common Rh antigens, common Kell antigens, or antigens of the Lewis, Kidd, or Duffy blood group systems. The majority of mAbs were of human origin but included a significant number of macaque mAbs, murine mAbs, and a small number of synthetically-designed recombinant antibodies. Both cellular (EBV-transformation, cell fusion) and molecular (phage display) approaches were used for antibody cloning. Analysis of certain groups of sequences demonstrated patterns of immunoglobulin gene restriction, repertoire shift, and somatic mutation. Analysis of other mAbs demonstrated the value of antibody sequence data for the design and production of novel reagents useful in blood group serology. PMID- 11889904 TI - In vivo studies of monoclonal anti-D and the mechanism of immune suppression. AB - Administration of anti-D immunoglobulin to D- women after delivery of a D+ infant has dramatically reduced the number of immunised women and cases of haemolytic disease of the fetus and newborn. The use of monoclonal anti-D might alleviate some of the pressures on maintaining adequate supplies of plasma sourced anti-D. Two human monoclonal antibodies, BRAD-3 (IgG1) and BRAD-5 (IgG3), with proven activity in in vitro functional (immunological) assays with cells bearing IgG Fc receptors (Fc gamma R) were selected for clinical studies. They were prepared by purification of IgG secreted by culture of the Epstein-Barr virus-transformed B cell lines in hollow fibre bioreactors. The clearance of D+ red cells injected into D- subjects was accelerated by prior injection of the monoclonal antibodies, both individually and blended (3:1, BRAD-5: BRAD-3). The subjects were protected from Rh D immunisation. A large multicentre study evaluated the BRAD-3/5 blend for its ability to prevent Rh D immunisation in 95 D- subjects given 400 micrograms i.m. 24 hours after injection of 5 ml D+ red cells. Challenge injections of D+ red cells alone were given 24 and 36 weeks later, and blood samples were taken every 4 weeks from the subjects throughout the study for detection of anti-D responses. There was one definite and one possible failure of protection; in one subject the plasma anti-D level rose from week 12 onwards, and in another individual rapid seroconversion was observed at week 28. Considering the relatively large dose of red cells and the number of subjects studied, it was concluded that the failure rate was much lower than in routine Rh D prophylaxis. The responder rate was 13% by week 36 and 24% by week 48. The low percentage of responders and the modest levels of endogenous anti-D produced suggested that administration of monoclonal anti-D had induced long-term specific suppression of anti-D responses in these subjects. The most likely mechanism of action was considered to be inhibition of B cells resulting from co-crosslinking antigen receptors with inhibitory Fc gamma R when the B cells contacted red cells that had bound passive anti-D. PMID- 11889905 TI - An Advisory Committee Statement (ACS). Committee to Advise on Tropical Medicine and Travel (CATMAT). Statement on travellers and rabies vaccine. PMID- 11889906 TI - The use of the Internet to inform young international travellers of contact with a case of meningococcal meningitis. PMID- 11889907 TI - Smallpox eradication: destruction of variola virus stocks. PMID- 11889908 TI - [Specific inhibitors of cyclooxygenase-2 (COX-2): current knowledge and perspectives]. AB - The Authors summarize the current knowledge on a new class of nonsteroidal anti inflammatory drugs (NSAIDs), the coxib (celecoxib and rofecoxib), in the treatment of rheumatic diseases. Celecoxib and rofecoxib are selective cyclooxygenase-2 (COX-2) inhibitors which possess the same anti-inflammatory and analgesic activities, but a better gastric tolerability compared to the non selective COX-1 and COX-2 inhibitors. The Authors also report other possible therapeutic effects of these NSADIs as evidenced by the more recent data of the literature. Celecoxib seems to reduce the incidence of new polyps in patients with familial adenomatous polyposis. It has been suggested the use of celecoxib as a protective drug against the development of colorectal cancer. Other (neoplastic) or pre-neoplastic conditions, such as bladder dysplasia, Barret esophagus, attinic keratosis and Alzheimer's disease seem to have benefit from this class of drugs. PMID- 11889909 TI - Myoepithelioma of the palate: a case report. AB - Myoepithelioma is a rare malignant neoplasm, representing about the 1% of all the salivary gland's tumors. It mainly affects women, particularly in the 7th and 8th decade of life. A case of myoepithelioma of the palate in a 70-year-old woman is described. The tumor was completely excised two years ago. Nowadays, there are no signs of local recurrence and/or distant metastasis. The discrepancy between the histological diagnosis of low-grade malignancy and the clinical aggressive behavior of the tumor is discussed. PMID- 11889910 TI - Radial artery in re-do coronary artery bypass grafting: our experience. AB - We evaluated our experience with the use of the radial artery as a key conduit in re-do coronary artery bypass surgery to determine the safety and efficacy and to compare this procedure to re-operations performed without the radial artery. Sixty-eight patients operated on re-do revascularization were studied: mean age was 67 years; 42 patients were in CCS III (62%) and 18 in CCS IV (26%); past myocardial infarction occurred in 12 patients (18%). We performed 116 anastomoses in all 68 patients (mean no. anastomoses/patient 1.7). Perioperative mortality was 4.4%. Three patients (4.4%) showed a transient postoperative low cardiac output syndrome; four (5.8%) had a respiratory failure and an acute renal failure occurred in 2 patients (2.9%). Four patients (5.8%) required re-operation for bleeding. The comparison of the radial re-do group (27 patients) with the non radial re-do group (41 patients) showed a lower mortality and morbidity in the former, even if p value was not significant. We conclude that the use of the radial artery in re-do coronary operations is safe, effective, allowing an additional conduit choice and may avoid late vein graft failure. PMID- 11889911 TI - Pericardial drainage for pericardial tamponade: surgical management criteria. AB - Aim of this study is the review of our experience in 82 patients treated by pericardial drainage for cardiac tamponade, to assess the efficacy and safety of different techniques and the related indications. The causes of pericardial effusion were: malignancy in 8 patients (9.7%), post-cardiac surgery in 12 (14.6%), while the others patients were admitted at our Institution with no identified preoperative diagnosis. Thirty-eight patients (46%) underwent subxiphoid pericardial drainage and 44 (54%) were operated on by catheter pericardiocentesis. There were no perioperative deaths. Two patients, who initially underwent pericardiocentesis, needed urgent sternotomy: the first patient developed a severe hypotension and bradicardia related to a vagal reaction and the other one because of accidental right ventricle puncture. Our experience indicates that subxiphoid pericardiocentesis provides expeditious, effective and durable treatment, with low morbidity, in case of pericardial effusions related to all causes. We believe that echocardiography is a powerful tool in the diagnosis and management of pericardial effusion. We conclude that pericardiocentesis seems to be the procedure of choice for patients with pericardial tamponade requiring an emergency treatment. PMID- 11889912 TI - Ischaemic mitral valve regurgitation: a surgical approach. AB - The aim of this study IS to determine surgical results after surgical mitral valve repair in ischaemic mitral regurgitation. MATERIALS AND METHODS: Between January 1999 and June 2000, 64 patients (5.1% of overall patients) underwent myocardial revascularization and mitral valve surgery. A Cosgrove-Edwards mitral annuloplasty ring was used in 59 cases (92.2%). Average patient age was 64.3 +/- 12.4 years (38 males, 21 females). Average degree of mitral regurgitation was 2.8 +/- 0.6. Average NYHA class was 3.5 +/- 0.5. Average ejection fraction (EF) was 40 +/- 12.5 percent. RESULTS: Post-operative 30-day mortality was 3.4% (2 patients). The follow-up was complete for 95 percent (mean 20.4 +/- 4.8 months for patients) and data showed an improvement of NYHA class (mean value 1.8 +/- 0.2) (p = 0.01) and ejection fraction (mean value 51.7 +/- 10.2) (p = 0.05) with residual mitral regurgitation value of 0.6 +/- 0.7. CONCLUSIONS: Mitral valve repair in coronary artery disease improves left ventricular function, quality of life and survival rate with low operative risk. Perioperative transesophageal echocardiography has a central role in surgical decision making. PMID- 11889913 TI - Reaction behaviors of glycine under super- and subcritical water conditions. AB - The influence of temperature and pressure on the dimerization and decomposition of glycine under simulated hydrothermal system conditions was studied by injecting a glycine solution into water in the sub- and supercritical state. The experiments at five different temperatures of supplied water--250, 300, 350, 374, and 400 degrees C--were performed at 22.2 and 40.0 MPa. At 350 degrees C, experiments under 15.0-40.0 MPa were conducted. Diglycine, triglycine (trace), diketopiperazine, and an unidentified product with a high molecular mass (433 Da) were the main products of oligomerization. The results show that temperature and pressure influence the extent of dimerization and decomposition of glycine. The maximum of dimers formation was observed at 350 and 375 degrees C at 22.2 and 40.0 MPa, respectively, and coincided with a high rate of glycine decomposition. Glycine, alanine, aspartic acid, as well as other amino acids, were obtained by injecting a mixture of formaldehyde and ammonia. The results support the oligomerization and synthesis of amino acids in a submarine hydrothermal system. PMID- 11889914 TI - Prebiotic oligomerization on or inside lipid vesicles in hydrothermal environments. AB - Oligomerization of amino acids proceeded on or inside lipid vesicles as a model of prebiotic cells in a simulated hydrothermal environment. When the suspension of lipid vesicles taking up monomeric glycine underwent a sudden temperature drop by traversing from a hot (180 degrees C) to a cold (0 degree C) region repeatedly while circulating through a closed reaction circuit, oligopeptides up to heptaglycine were formed even in the absence of condensing agents. PMID- 11889915 TI - Coding rules for amino acids in the genetic code: the genetic code is a minimal code of mutational deterioration. AB - Coding rules for amino acids in the genetic code are discussed from the point that the genetic code is a minimal code of mutational deterioration. The global mutational deterioration (GMD) function is defined through several parameters describing single base mutations and amino acid distances. The problem of searching for the global minimum of the GMD function is discussed in some detail. From GMD minimization under initial constraints we have succeeded in deducing the standard genetic code. PMID- 11889916 TI - Primordial coding of amino acids by adsorbed purine bases. AB - Scanning tunneling microscopy and chromatography experiments exploring the potential templating properties of nucleic acid bases adsorbed to the surface of crystalline graphite, revealed that the interactions of amino acids with the bare crystal surface are significantly modulated by the prior adsorption of adenine and hypoxanthine. These bases are the coding elements of a putative purine-only genetic alphabet and the observed effects are different for each of the bases. Such mapping between bases and amino acids provides a coding mechanism. These observations demonstrate that a simple pre-RNA amino acid discrimination mechanism could have existed on the prebiotic Earth providing critical functionality for the origin of life. PMID- 11889917 TI - Europa as an abode of life. AB - Life as we know it on Earth depends on liquid water, a suite of 'biogenic' elements (most famously carbon) and a useful source of free energy. Here we review Europa's suitability for life from the perspective of these three requirements. It is likely, though not yet certain, that Europa harbors a subsurface ocean of liquid water whose volume is about twice that of Earth's oceans. Little is known about Europa's inventory of carbon, nitrogen, and other biogenic elements, but lower bounds on these can be placed by considering the role of commentary delivery over Europa's history. Sources of free energy are challenging for a world covered with an ice layer kilometers thick, but it is possible that hydrothermal activity and/or organics and oxidants provided by the action of radiation chemistry at Europa's surface and subsequent mixing into Europa's ocean could provide the electron donors and acceptors needed to power a Europan ecosystem. It is not premature to draw lessons from the search for life on Mars with the Viking spacecraft for planning exobiological missions to Europa. PMID- 11889918 TI - Synaptic long-term depression (LTD) in vivo recorded on the rat cerebellar cortex. AB - Long-Term Depression (LTD) of the parallel fiber synapses of the cerebellar cortex has been intensively studied over the last 20 years and is now considered to be a physiological mechanism underlying learning and memory of the cerebellar cortex. With microelectrode recording in vivo, the induced LTD is recorded reliably up to 2 hours. Using surface electrodes we have recorded parallel fiber responses due to the currents generated by the AMPA type receptors of the dendritic spines in the intact vermal cortex of decerebrated rats. We have found that by conjunctively stimulating the climbing and parallel fiber pathways, an LTD was induced which persisted for as long as the recording conditions permitted. The longest lasting LTD of our present results was for 5 hours. PMID- 11889919 TI - Time courses of aspartate and glutamate concentrations in the focus area during penicillin induced epileptiform activity in awake rats. AB - 1. In 16 awake rats, the time courses of cortical aspartate (ASP) and glutamate (GLU) concentrations were investigated before, during and after penicillin induced epileptiform activity (PCN-EA). The amino acids were collected by means of the microdialysis technique from a site within the motor cortex located close to the PCN focus. PCN-EA was recorded by means of 12 cortical electrodes. 2. The median time courses of ASP and GLU concentrations demonstrated no relation to the time course of PCN-EA. In contrast, the analysis of individual time courses revealed three response types for both ASP and GLU which could not be related to specific sites within the motor cortex. 3. The ASP concentration was i) not affected by epileptiform activity (8 of 16 rats), ii) changed synchronously with the PCN-EA (3 of 16), or iii) increased transiently or in a long-lasting manner during or after disappearance of PCN-EA (5 of 16). 4. The GLU concentration was i) not effected or only slightly affected by epileptiform activity (5 of 16), ii) decreased strongly during the activity (7 of 16), or iii) increased during the epileptiform activity (4 of 16). 5. If all rats were taken into account, the overall observation was absence of a uniform correlation type between the time courses of ASP and GLU concentrations; i.e. epileptiform activity affects the time courses of ASP and GLU concentrations in an independent manner. 6. These results demonstrate that consideration of median values alone can mask significant effects of epileptiform activity on the biochemistry of the brain in individual animals. The study also shows that a consideration of individual time courses of physiological and biochemical parameters is mandatory for the understanding of basic mechanisms of epilepsy. The results support the hypothesis about the activation of neuroprotective mechanisms by epileptiform activity. PMID- 11889920 TI - Comparative study of the epileptogenic effect of kainic acid injected into the cerebral cortex, hippocampus and amygdala in adult cats chronically implanted. AB - Intracerebral injection of kainic acid in cerebral cortex, hippocampus or amygdala in cats chronically implanted showed that: 1) Hippocampus and amygdala presented a greater sensitivity than the cerebral cortex, while hippocampus presented a greater sensitivity than the amygdala to the generation of an epileptic focus. 2) Comparison of latency, mean duration of afterdischarges, and the mean time period to obtain the peak intensity of the afterdischarge in the three cited structures, showed that mean latency of the first afterdischarge was significantly shorter in hippocampus and amygdala compared with the cerebral cortex. Moreover the mean time period to reach the peak intensity of the afterdischarge was again shorter in the subcortical structures. 3) The epileptic foci both in hippocampus and amygdala were blocked by CNQX and muscimol. 4) The behavioral changes depended on the intensity of the epileptic process. Tonic clonic convulsions appeared only when the motor cerebral cortex was involved. Finally, 5) kainic acid injections in hippocampus and amygdala elicited an intense neuronal destruction and gliosis of these structures. We conclude that intracerebral injection of low doses of kainic acid in cats represent a good model to study focal epileptic thresholds in the CNS. PMID- 11889921 TI - Induction and blockade of epileptic foci by intracerebral injection of glutamatergic agonists and antagonists in frerly moving cats. AB - The aim of the present work was to test in adult cats the capability of three glutamatergic agonists, NMDA, AMPA and ACDP as epileptogenic agents. Drugs were microinjected in amygdala or hippocampus, and once generated an epileptic focus three selective glutamatergic antagonists NMDA, CNQX and MCPG, were tested. Before and after injection both the EEG and the behavior were continuously monitored. The results were as follows: 1) AMPA showed a greater capability than NMDA or ACPD to generate a chronic epileptic focus; 2) AMPA elicited a greater epileptogenic effect in hippocampus than in amygdala; NMDA had similar epileptogenic effect in both cited structures, and ACPD had not effect; 3) of the three glutamatergic antagonists used to block a long lasting focus, the most effective one was CNQX, which showed a greater effect in hippocampus than in amygdala; 4) comparison between the epileptogenic effect of AMPA and kainic acid (first paper) in the same structure, showed that kainic acid is about 15 fold more epileptogenic. A discussion of the probable mechanisms of these results was undertaken. PMID- 11889922 TI - Capsaicin-sensitive muscle afferents modulate the monosynaptic reflex in response to muscle ischemia and fatigue in the rat. AB - The role of muscle ischemia and fatigue in modulating the monosynaptic reflex was investigated in decerebrate and spinalized rats. Field potentials and fast motoneuron single units in the lateral gastrocnemious (LG) motor pool were evoked by dorsal root stimulation. Muscle ischemia was induced by occluding the LG vascular supply and muscle fatigue by prolonged tetanic electrical stimulation of the LG motor nerve. Under muscle ischemia the monosynaptic reflex was facilitated since the size of the early and late waves of the field potential and the excitability of the motoneuron units increased. This effect was abolished after L3-L6 dorsal rhizotomy, but it was unaffected after L3-L6 ventral rhizotomy. By contrast, the monosynaptic reflex was inhibited by muscle fatiguing stimulation, and this effect did not fully depend on the integrity of the dorsal root. However, when ischemia was combined with repetitive tetanic muscle stimulation the inhibitory effect of fatigue was significantly enhanced. Both the ischemia and fatigue effects were abolished by capsaicin injected into the LG muscle at a dose that blocked a large number of group III and IV muscle afferents. We concluded that muscle ischemia and fatigue activate different groups of muscle afferents that are both sensitive to capsaicin, but enter the spinal cord through different roots. They are responsible for opposite effects, when given separately: facilitation during ischemia and inhibition during fatigue; however, in combination, ischemia enhances the responsiveness of the afferent fibres to fatigue. PMID- 11889923 TI - Vestibular mechanisms involved in idiopathic scoliosis. AB - Patients affected by idiopathic scoliosis (IS) show not only a spinal deformity, but also postural and oculomotor deficits suggesting that such syndrome can be related to a vestibular disfunction. It appears, however, that, in children, a slight unbalance in the activity of vestibular complex of both sides escapes the neuronal mechanisms responsible for vestibular compensation and leads to the spinal curvature which characterises IS. Such process could be reinforced by a disrupted integration of vestibular and visual signals at cortical level, leading to an altered perception of the vertical and to abnormal motor commands. In addition to the classical ascending and descending pathways arising from the vestibular nuclei, which utilize glutamate or GABA as neurotransmitters, labyrinthine afferents may also affect spinal, cerebellar and cerebrocortical structures, through the noradrenergic and serotoninergic systems, which originate from the locus coeruleus and the raphe nuclei, respectively. Due to the role of these neuromodulators in brain plasticity, a disruption in the activity of monoaminergic neurons could favour the development of postural and oculomotor deficits. An impaired release of monoamine at cerebrocortical level could also explain the cognitive deficits which may occur in IS patients. PMID- 11889924 TI - Florida is leader in self-funded dental plans. PMID- 11889925 TI - Board of Dentistry adjustments would benefit patients and dentists. PMID- 11889926 TI - Testing your diagnostic skills #57. Case No. 1. Denture sore spots. PMID- 11889927 TI - Testing your diagnostic skills #57. Case No. 2. Central giant cell granuloma. PMID- 11889928 TI - Gonna git me a laser. PMID- 11889929 TI - Handling a difficult personality. PMID- 11889931 TI - Automated external defibrillators and liability. PMID- 11889930 TI - Coding for routine periodontal treatment: Part 2. PMID- 11889932 TI - Dr. Allan Jacobs: Endodontist and Oakland County Trustee. Interview by Jeff Mertens. PMID- 11889933 TI - Oral pathology quiz #3. Cleidocranial dysplasia. PMID- 11889934 TI - [Particular profile of the zymodemes of Leishmania infantum causing visceral leishmaniasis in Tunisia]. AB - The isoenzymatic typing of 16 stocks of Leishmania, isolated from Tunisian visceral leishmaniasis (VL) cases, revealed that all strains belonged to Leishmania (L.) infantum species. Although zymodeme MON-1 was predictably the most frequent (9 cases), it came as a surprise that L. infantum MON-24 was responsible of about third of cases. This latter zymodeme, while previously reported in Tunisia, Algeria and Spain, was assumed to be a dermotropic and the few cases of VL that it caused occurred always in HIV infected patients. L. infantum MON-80, occasionally reported during both cutaneous and VL of immunocompetent infants was identified in 2 patients. This report confirms that in addition to the more common L. infantum MON-1, zymodeme MON-24 has a substantial role in generating VL in immunocompetent infants in Tunisia. PMID- 11889935 TI - [Diagnostic, therapeutic and prognostic aspects of intestinal typhoid perforations in children of Abidjan, Cote d'Ivoire]. AB - We conducted a retrospective study on 48 children (with a sex ratio of 22 males to 26 females) who had been operated over the preceding 10 years for typhoid perforation on the viscera ward within the paediatric surgery department of Yopougon teaching hospital in Abidjan, Cote d'Ivoire. The mean age was 9 years 3 months ranging from 3 to 16 years. Typhoid peritonitis was diagnosed on the basis of symptoms, intestinal injuries and only rarely by way of biological examinations. Medical treatment associated three antibiotics: aminosid, metronidazol and the third generation of cephalosporin. Hydroelectrolytical and haematological resuscitation was performed 3 to 6 hours before laparotomy. Excision-suture was made in 81% of cases. Exclusive parenteral nutrition began 48 hours after the laparotomy. Mortality occurred in 6% of patients and morbidity in 46%. Complications were parietal suppurations, digestive fistula, parietal hernia, early occlusive syndromes and necrosed cholecystisis. Rectorragy and leucocytosis were considered as pejorative signs when associated to the classic typhoid peritonitis. PMID- 11889936 TI - [Rare but dangerous foreign body of the external auditory canal: alkaline batteries]. AB - About an observation in a 3 year-old child of the alkaline drum kits in the external acoustic lacunae, the authors underline that they are rare but dangerous. The mechanisms of toxic activity of these piles are exposed, as well as the complications that they engender and the principles of their treatment. The ablation should be carried out as quickly as possible. The excision is usually possible by natural pathways. However, attention should be paid to preventing post-operational stenosis by using gauze and administering careful local treatment. PMID- 11889937 TI - [Epidemiological aspects of Burkitt's lymphoma in children of Madagascar]. AB - RATIONALE: Burkitt lymphoma (LB), a frequent, very progressive cancer with multiple factors, can be cured. However, the mortality rate remains high in Madagascar. OBJECTIVE: To analyse the epidemiological aspects of LB as well as related socio-economical issues in order to improve successful treatment of the disease. METHODS: Retrospective study of files for children aged under 15 years, hospitalised for LB in the Antananarivo oncology unit from October 1985 to June 2000. The inclusion criteria were anatomo-pathological, clinical and/or X-ray results. Studied parameters included age, sex, ethnic group, medical history, and the distance covered by the child before his/her hospitalisation. FINDINGS: The 77 cases of LB represented 16% of all children aged under 15 years seen in the hospital. The characteristics of the cases corresponded to those of African endemic LB. Most of the children with LB came from areas with endemic malaria, the Eastern and the Centre of Madagascar. All of them belonged to underprivileged families. Early medical advice was sought but distance from services delayed treatment. Various units referred the children, but especially oral surgeons (stomatologists) and ORL physicians. CONCLUSION: A strategy to ensure rapid treatment for children suffering from LB should be developed, from their region of origin up until treatment. This should involve parents as well as all members of the medical staff in charge of these children. PMID- 11889939 TI - [Leptospirosis in children of Libreville: difficult diagnosis, apropos of 1 case]. AB - Leptospirosis is a widespread zoonosis, which is diagnosed less frequently in children than might be expected from the level of exposure to hazards, especially in tropical areas. A 15 1/2-year-old Gabonese boy was admitted following five days of fever, headache, myalgia, abdominal pain, diarrhea, intestinal bleeding, jaundice and conjunctival suffusion. Laboratory data showed abnormal liver and renal function tests, and diagnosis of Plasmodium falciparum malaria was confirmed by thin blood smear. The patient did not clinically improve despite antimalarial treatment and then leptospirosis was suspected. Serologic tests were performed and leptospirosis was later confirmed. Antibiotic treatment (cefuroxim) was given. The outcome was good, liver and renal tests returned to normal in a few days. In tropical area, leptospirosis should be considered in children who are diagnosed with either an unexplained fever, a pseudo-influenza syndrome, or jaundice with hepatorenal involvement and gastrointestinal bleeding. PMID- 11889938 TI - [Burkitt's lymphoma in children of Madagascar. Anatomo-clinical forms, therapeutic and evolutive aspects]. AB - RATIONALE: Burkitt Lymphoma (LB), a very progressive malignant lymphoma, can now be cured by chemotherapy. However, protocols used currently by developed countries are costly and can cause problems of tolerance for underprivileged children. OBJECTIVE: To develop a protocol of care for Malagasy children suffering from LB. METHODS: A retrospective study of the files of children aged under 15 aged years, hospitalised for LB with anatomo-pathological evidence, in the Antananarivo Oncology Unit from October 1985 to June 2000. Clinical characteristics, (paraclinical) investigations, treatment and responses to treatment were studied. FINDINGS: 40 medical records included a LB anatomo pathological evidence. The mean age of children was 7.5 years, with a sex ratio in favour of males. All children were underweight. Maxillo-facial tumours prevailed. Other locations for tumours had also been observed, both unique or immediately multiple. Patients seem to have arrived at the hospital in the early stages of the disease, but lack of investigation probably introduced biases to the evaluation of these stages. Chemotherapy, even though incomplete due to its cost, remained the main means of treatment. Generally speaking, an immediate, favourable response was obtained, but the toxicity of chemotherapy, especially haematological toxicity, contributed to malnutrition. Overall results indicated an immediate mortality rate of 22.5%, but there was loss of follow-up for many patients. CONCLUSION: A specific protocol of LB care in Madagascar appears to be possible. Such a protocol should be based on experience with treatments, and should take into account disease characteristics, the response of Malagasy children to the treatment, as well as the country's economic state. PMID- 11889940 TI - [Evaluation of likely antibiotherapy in bacterial-like acute pneumopathies in patients hospitalized in Africa]. AB - We conducted a retrospective analysis of 100 records of adult African patients hospitalised for bacterial-like acute pneumonia. The objective of the study was to evaluate the use and efficacy of probabilist antibiotherapy. The study population was made up of 57% men and 43% women. Serious clinical symptoms were found in 31% of the patients, with serious x-ray and biological anomalies for respectively 67% and 51% cases. Secondary morbidity was associated with pneumonia in 30% cases. In the first intention, the three (3) most prescribed antibiotics are beta-lactamins (84%), fluoroquinolons (25%), and aminosids (25%). Sulfamids, macrolids and imidazols were prescribed together in 18% cases. Monotherapy was prescribed in 53% cases and concerned especially amoxicillin (39/53) and fluoroquinolons (5/53). Double therapy was used in 42% of cases and consisted of amoxicillin + aminosid (21/42) and amoxicillin + fluoroquinolon (17/42). Three antibiotics were noted for 5 cases. The intravenous administration was frequently used (68%), either alone (27%), either associated with other modes of drug administration (41%). Mean duration of antibiotherapy was 12.71 days. 73% of patients improved, 22% failed to improve and 5% died. Antibiotherapy was influenced by the seriousness biological signs and by the mode of administration of antibiotherapy in monotherapy. Deaths occurred precociously and concerned HIV positive (4/5) patients presenting at least 2 factors of co-morbidity and having received beta-lactamin in monotherapy. PMID- 11889941 TI - [Salubrity of ice sold in blocks: from production to sales]. AB - We conducted a study in order to determine the salubrity of ice produced in blocks, tracing them from 10 factories in Abidjan to 10 different market places where they are sold in town. On the basis of physico-chemical and microbiological analyses, the water was shown to be potable. However, the results of the analysed samples taken from the ice indicated a deterioration of the physico-chemical and microbiological qualities. The data revealed that consumers may be exposed to water-borne diseases such as cholera, salmonelloses, shigelloses, gastro enteritis. Ice producers and vendors must be provided with special training in sanitation so as to prevent the transmission of pathogenic bacteria in ice. PMID- 11889942 TI - [Toxoplasmosis, rubella, syphilis, hepatitis B and HIV infection in women being followed for pregnancy in a population on the east coast of New Caledonia ]. AB - In view of the sparse available data concerning the main infectious illnesses screened for during pregnancy (toxoplasmosis, rubella, syphilis, hepatitis B virus and HIV) in the Northern Province and generally throughout New Caledonia, we decided to undertake a retrospective study based on the files of pregnant women having consulted between September 1996 and September 1999 in the East Coast provincial hospital (New-Caledonia). First, we wished to estimate the prevalence of toxoplasmosis, rubella, syphilis, hepatitis B virus and HIV Second, we wished to specify the main characteristics of these pregnant women, to trace the evolution of seroprevalence over the study time period and consider the influence of various factors such as age, place of residence and professional occupation. The biological study of these pregnancies was hindered by the fact that approximately half of women consult only from the second trimester of their pregnancy. This result underlines the importance of emphasising efforts aiming to bring women to consult earlier and of improving the regularity and also the interpretation of serological tests (especially for toxoplasmosis serology). Within this population, we found the following percentages of seroprevalence: for toxoplasmosis between 83.6% and 89.6% (zone of hyperendemla), for rubella between 91.6% and 95.8%, for syphilis between 7% and 12.4%, for hepatitis B virus between 61.8% and 76% (for women immune to hepatitis B, the frequency of acquisition of immunity by viral infection was understood between 64.3% and 80.3%) and for HIV 0%. The hepatitis B surface antigen (HbsAg) carrier rate was estimated between 1.7% and 4.9%. Following and informing pregnant women of the risk of toxoplasmosis appears to be of key importance as well as screening HbsAg carriers in order to limit viral transmission to the foetus. PMID- 11889943 TI - [Neurological disorders and endemic goiter: comparative analysis of 2 provinces in Togo]. AB - Neurological diseases are one of the main causes of high mortality and morbidity in developing countries. The prevention of these diseases is possible if they are well known. The purpose of this study was to evaluate the prevalence and the distribution of the main neurological affections in two rural districts. WHO research protocol for measuring the prevalence of neurological disorders in developing countries was used by the same staff. The first survey was conducted in Kloto from July to August 1989 on 19,241 inhabitants and the second in Akebou from January to February 1995 on 4182 subjects. Akebou was known as endemic goiter district. The prevalence of epilepsy was 12.3@1000 in Kloto and 13.1@1000 in Akebou. That of stroke was 2.4@1000 in Kloto versus 1.7@1000 in Akebou. The prevalence of Parkinson's disease was 0.2@1000 in Kloto and 0.2@1000 in Akebou. That of psychomotor retardation was 3.2@1000 in Kloto versus 8.1@1000 in Akebou. The prevalence of neurological cretinism was 97@1000 while that of myxoedematous cretinism was 31@1000 in Akebou. Apart from neurological disease, the prevalence of goitre was 43% in females and 26.1% in males in Akebou. Mental retardation and cretinism were frequent in Akebou, a district endemic for goitre. Besides stroke and febrile convulsions which predominated in Kloto, the main neurological disorders were most prevalent in Akebou. Community based care for epilepsy is provided in these districts as is the prevention of iodine deficiency in the Akebou district with the intention of reducing neurological affections. PMID- 11889944 TI - [Microbiological quality of " ice, ice cream. sorbet" sold on the streets of Phnom Penh; April 1996-April 1997]. AB - A study of the microbiological quality of ice lice creams/sorbets sold on the streets of Phnom Penh city was conducted from April 1996 to April 1997. Socio demographic and environmental characteristics with two ice/ice creams samples were collected from vendors selected in the city. A total of 105 vendors and 210 ice/ice creams samples were randomly selected for the study period. Ice/ice cream vendors in the streets of Phnom Penh were adults (mean age: 28 years old) with a male predominance (86.5%). Mean educational level of vendors was 5 years with no training in mass catering. Most ice creams and sorbets (81.7%) were made using traditional methods. Microbiological analysis performed in the laboratory of Pasteur Institute of Cambodia indicated the poor bacteriological quality of the samples. The proportions of samples classified unsafe according to microbiological criteria were 83.3% for total bacterial count at 30 degrees C, 70% for total coliforms, 30% for faecal coliforms, 12.2% for Staphylococcus aureus and 1.9% for presence of Salmonella spp. These bacterial results suggest that many other food products sold in the streets may be similarly poor. Safety measures should be undertaken to avoid potential threats. Regulation of the street food sector should be part of a larger strategy for enhanced food safety and environmental quality in the city. PMID- 11889945 TI - [Incidence of HIV infection in consultants reviewed after a first negative test in an anonymous and free screening center at the Institut Pasteur of Cambodia, 1996-1999]. AB - A retrospective study was performed to determine the incidence of HIV seroconversion among repeat consultants attending the voluntary testing and counselling centre of the Institut Pasteur of Cambodia as well as factors associated with HIV seroconversion. From 1996 to 1999, 5541 repeat consultants were selected for the study. Exclusion criteria included being aged under 15 years, having initially tested positive or inconclusive and a time span of fewer than 30 days since the last test. In all, 276 persons had seroconverted to HIV, giving an incidence rate of 5.56 per 100 person-years. The seroconversion rate declined from 8.46% in 1996, to 3.06% in 1999 (chi 2 test for trend, p = 10(-5)). Among the risk factors analysed, 3 were significantly associated with lack of seroconversion: being a student (RR = 0.53, p = 0.032) or a civil servant (RR = 0.63, p = 0.012) and systematic condom use with causal partners (RR = 0.37, p = 10(-5)). The decline of HIV seroconversion among repeat consultants attending the VCT centre over the study period may reflect changes in risk behaviour and the beneficial impact of counselling. PMID- 11889946 TI - [Visceral leishmaniasis in Cameroon. Seroepidemiologic survey in the Kousseri region, north Cameroon]. AB - A sero-epidemiological survey of school children was carried out in Kousseri, a focus for visceral leishmaniasis. Sero-immunological assays for the detection of anti-Leishmania antibodies were based on the indirect immunofluorescence assay test and counter-immunoelectrophoresis. 9 out of 223 school children tested positive for visceral leishmaniasis (seroprevalence rate of 4%). These 9 cases had no history of the disease. The data obtained confirm the endemicity of visceral leishmaniasis in this focus and call for extensive studies in order to determine the prevalence of the disease in the entire population as well as the main components of the transmission cycle. PMID- 11889947 TI - Extraction of physiological and clinical information from intra-atrial electrograms during atrial fibrillation: review of methods. AB - This paper presents a review of the evolution of methods and algorithms for the analysis of intra-atrial recordings, with special emphasis to the extraction of physiological and clinical information during atrial fibrillation. The principal time-domain and frequency-domain methods of electrogram analyses are described, and their physiological interpretation and clinical applications are discussed. In addition, the recent findings from complex system theory and chaos theory approaches are highlighted. PMID- 11889948 TI - Non-linear time series analysis: methods and applications to atrial fibrillation. AB - We apply methods from non-linear statistical time series analysis to characterize electrograms of atrial fibrillation. These are based on concepts originating from the theory of non-linear dynamical systems and use the empirical reconstruction density in reconstructed phase space. Application of these methods is not restricted to deterministic chaos but is valid in a general time series context. We illustrate this by applying three recently proposed non-linear time series methods to fibrillation electrograms: 1) a test for time reversibility in atrial electrograms during paroxysmal atrial fibrillation in patients; 2) a test to detect differences in the dynamical behaviour during the pharmacological conversion of sustained atrial fibrillation in instrumented conscious goats; 3) a test for general Granger causality to identify couplings and information transport in the atria during fibrillation. We conclude that a characterization of the dynamics via the reconstruction density offers a useful framework for the non-linear analysis of electrograms of atrial fibrillation. PMID- 11889949 TI - Discrimination of atrial rhythms by linear and non-linear methods. AB - The discrimination among atrial rhythms is obtained through the analysis of linear and non-linear dynamics of atria electrograms and local atrial period (LAP) series. Linear and non-linear degree of coupling between atrial electrograms have been assessed using cross-correlation (CCI) and synchronization indexes. Dynamics of LAP series were investigated using level of predictability (LP) and regularity indexes. Atrial fibrillation (AF) episodes, classified according to Wells' criteria, were investigated. We found that metrics obtained from LAP series provide the best performance in the classification of AF rhythms. The LP index misclassified only 12 AF episodes over 87. Synchronization index was the most performing metric in the discrimination between organized and not organized fibrillation: detection of organized rhythm; sensitivity (SE) > 95%; positive predictability (+P) > 95%. LP (SE: 89%; +P: 89%) and CCI (SE: 87%; +P: 83%) provide slightly lower performances. PMID- 11889950 TI - Non-invasive assessment of atrial fibrillation (AF) cycle length in man: potential application for studying AF. AB - Non-invasive assessment of the fibrillatory frequency of atrial fibrillation (AF) can be performed by frequency domain analysis. The peak frequency in the derived spectrum can be converted to a dominant atrial cycle length (DACL). The DACL can be altered through autonomic modulation or pharmacologic manipulation, but the change in DACL is less marked in those with a short DACL value. In patients with AF, those with a short duration of the arrhythmia have longer DACL values. Finally, patients with paroxysmal AF generally exhibit longer DACL values than patients with permanent AF. Thus non-invasive assessment of the atrial fibrillatory cycle length provides a useful index of atrial refractoriness and has the potential of clinical utility in patient assessment and treatment planning. PMID- 11889951 TI - Frequency domain analysis of endocardial signals. AB - Frequency domain analysis, applied to atrial endocardial electrograms during atrial fibrillation, has been used to develop automated arrhythmia detection schemes for implantable devices and to investigate electrophysiologic mechanisms. Such analysis may be used to quantify both temporal and spatial organization during atrial fibrillation. Specifically, autopower spectra and coherence spectra reveal electrogram characteristics that are powerful in discriminating atrial fibrillation from sinus rhythm and regular atrial tachycardias. Furthermore, changes in spectral characteristics with drug administration reveal nonstationarities in spatial organization and in underlying electrophysiologic mechanism. The usefulness of frequency domain analysis in the study of atrial fibrillation is influenced by electrode configuration, the method of spectral estimation and other clinical variables. PMID- 11889952 TI - Measures of organization during atrial fibrillation. AB - Atrial fibrillation, a rhythm classically described as totally disorganized, is now recognized to have some structure in its activation. Even though that structure may be very complex, it clearly exists, and researchers continue to try to quantify that structure. One problem with this quantification is that "organization" is an ambiguous term that can have many interpretations. Rather than attempt to impose a particular definition of "organization," this review categorizes the methodologies for quantifying atrial fibrillation organization based on the number of recording channels used in the methods. This method of categorization is not only convenient, but is also descriptive of the different "philosophical" definitions of organization that various researchers have. PMID- 11889953 TI - Deterministic patterns and coupling of bipolar recordings from the right atrium. AB - The presence of organization of atrial activation processes during atrial fibrillation has been investigated to assess whether the activation sequences are wholly random or are governed by deterministic mechanisms. We performed a linear analysis based on the cross correlation function and a non linear analysis based on two algorithms. The first one detects and qualifies recurrent patterns of the coupling between atrial period time series by means of the recurrence plot quantification approach; the second one estimates the non linear coupling between activation sequences in two sites following a multivariate embedding approach. The detection of spatio-temporal recurrent patterns and of non linear coupling together with the surrogate data results indicate that during atrial fibrillation a certain degree of local organization exists, likely caused by deterministic mechanisms of activation. PMID- 11889954 TI - Effect of atrial fibrillation organization on internal defibrillation threshold. AB - Aim of the present study is to assess the effect of the organization of atrial fibrillation in the right atrium on the minimum effective cardioversion energy. Thirteen patients were studied, selected a priori for low-energy internal cardioversion (LEIC). LEIC was carried out at increasing energy levels. All patients were monitored by a multipolar basket catheter in basal condition and during cardioversion. Our results showed that the mean organization correlates with the energy of successful cardioversion, and that after each shock no significant differences (Kruskal-Wallis test) were observed in the level of organization. Similar results have been obtained for the differences of organization values, immediately before and after each shock. These results prompt for further studies aimed at measuring electrophysiological properties to predict energy thresholds. PMID- 11889955 TI - Mapping of atrial fibrillation. AB - Cardiac mapping has been defined as: "a method by which cardiac signals are recorded from multiple sites of the heart and spatially depicted as a function of time in an integrated manner". It requires determination of the local activation time at each electrode and the creation of activation maps which provide a spatial model of the activation sequence. With respect to atrial fibrillation, mapping is useful to gain insight into the underlying mechanism of atrial fibrillation. In this review, we will discuss the mapping studies of experimental and clinical atrial fibrillation. PMID- 11889956 TI - Atrial mapping by basket catheter in patients with atrial fibrillation submitted to linear ablation. AB - Recently non-pharmacological therapies for atrial fibrillation (AF) have been developed. The electrophysiological mechanisms of AF is thought to be the development of multiple reentrant wavelets circulating around anatomic barriers and variable regions of functional conduction block responsible of the perpetuation of the arrhythmia. Also the role of the triggering foci has been highlighted. To cure AF by means of non pharmacological therapy we may eliminate and/or modify the substrate. To better understand the mechanism underlying the AF and to choose the best ablation strategy is of fundamental importance to map the right and the left atrium during AF. Our experience shows that in chronic idiopathic AF disorganized atrial activity is observed at all atrial regions while in paroxysmal idiopathic AF the left septum and the right atrial posterior areas are highly disorganized while the lateral region shows more organized atrial electrical activity. Multipolar basket catheters are extremely useful in mapping right and left atrium in order to guide the best ablation strategy. PMID- 11889957 TI - Pulmonary vein isolation by circumferential radiofrequency lesions in atrial fibrillation. From substrate to clinical outcome. AB - Circumferential ablation around pulmonary vein ostia by CARTO system was performed in 98 patients with paroxysmal and 29 with permanent atrial fibrillation (AF). Preablation and postablation activation, propagation and voltage maps were obtained. A total of 135 +/- 18 radiofrequency pulses were delivered. After a follow-up of 14.7 +/- 3.3 months, 84 patients with paroxysmal and 22 with chronic AF are in sinus rhythm and 75 of them without antiarrhythmics. Only the area extent of low voltage potentials within and just around the lesions distinguished patients with and without successful ablation. Pulmonary vein isolation is an effective procedure to cure resistant AF; the extent of lesion area around pulmonary vein ostia may be crucial in predicting the outcome. PMID- 11889958 TI - Algorithmic complexity. A new approach of non-linear algorithms for the analysis of atrial signals from multipolar basket catheter. AB - Symbolic dynamics as a non linear method and computation of the normalized algorithmic complexity (C alpha) was applied to basket-catheter mapping of atrial fibrillation (AF) in the right human atrium. The resulting different degrees of organisation of AF have been compared to conventional classification of Wells. Short time temporal and spatial distribution of the C alpha during AF and effects of propafenone on this distribution have been investigated in 30 patients. C alpha was calculated for a moving window. Generated C alpha was analyzed within 10 minutes before and after administration of propafenone. The inter-regional C alpha distribution was statistically analyzed. Inter-regional C alpha differences were found in all patients (p < 0.001). The right atrium could be divided in high and low complexity areas according to individual patterns. A significant C alpha increase in cranio-caudal direction was confirmed inter-individually (p < 0.01). The administration of propafenone enlarged the areas of low complexity. PMID- 11889959 TI - Clinical evaluation of disorganization during atrial fibrillation as a guide to radiofrequency ablation. AB - Aim of this study was to investigate the spatial patterns of organization of fibrillation in the right atrium using bipolar recordings from a basket catheter. Study population consists of 17 patients with persistent atrial fibrillation (AF), selected a priori for low energy internal cardioversion. Organization was measured by the number of occurrences--i.e. the percentage number of points laying on the baseline--which has been demonstrated to match Wells' criteria for the classification of AF. Well-defined and stable patterns of organization characterize the electrical activity of the right atrium in these patients. The organization in the entire right atrium showed an individual rather than common distribution. This may have implications in the choice of regions candidate for ablation. PMID- 11889960 TI - Different atrial regional patterns of activation during atrial fibrillation: is there any relationship with the anatomy? AB - Right atrial (RA) mapping has been recently more carefully examined in patients with idiopathic atrial fibrillation (AF) in order to improve radiofrequency (RF) catheter-mediated ablation lines to control recurrences. The aim of this study was to map right atrial activation during AF to analyze relationship between anatomy and atrial activation for specific sites. Twenty-four patients with recurrent, drug-refractory, paroxysmal AF underwent an extensive mapping of the RA before attempting RF linear lesion catheter ablation. A typical pattern of atrial activation was recorded in all patients which was consistent with a more regular activity on the trabeculated right atrium (type I AF) and a more fragmented and complex activation on the posterior and the anterior septum (type II and III AF). This paper helps to understand the influence of the anatomic barriers to atrial activation during atrial fibrillation. PMID- 11889961 TI - Repolarization and refractoriness in patients with persistent atrial fibrillation. AB - The use of monophasic action potential (MAP) recordings has been of important value during atrial fibrillation to understand the possibility of local pacing capture during the arrhythmia, while MAP and refractoriness determination after sinus rhythm restoration have highlighted the issue of electrophysiological remodeling owing to rate. Moreover the contemporary recording of MAP and refractoriness at the same atrial sites permitted to better understand the behavior of the ERP/MAP ratio in these patients. Local atrial pacing capture has been demonstrated in humans with chronic atrial fibrillation and suggests the presence of re-entrant circuits with large excitable gaps. The studies about atrial remodeling have shown a shortening of atrial ERP or monophasic action potential duration after cardioversion of persistent AF, while discordant results have been observed for what it concerns refractoriness adaptation to rate. Finally, the recording of a mean ERP/MAP90 ratio < 1 at all the pacing cycle lengths, indicates that no post-repolarization refractoriness was present after cardioversion of persistent atrial fibrillation. PMID- 11889962 TI - Clinical, electrocardiographic and electrophysiological predictors of atrial fibrillation development in different cardiac substrates. AB - We performed a review of the current literature in order to evaluate clinical, electrocardiographic and electrophysiologic parameters predictive of atrial fibrillation development. Clinical parameters were obtained from two large observational studies (the Framingham heart study and the Cardiovascular health study). Different laboratoristic predictors were also reviewed: ECG-derived predictors, among which we separately evaluated those derived from the 12-lead surface ECG and those derived from the signal averaged P-wave, and other electrophysiologic predictors as atrial monophasic action potential analysis. We also evaluated the clinical value of these different parameters in atrial fibrillation in patients with no overt structural heart disease and in the most common clinical conditions known to be related to atrial fibrillation development such as hypertension, heart failure, cardiovascular surgery. PMID- 11889963 TI - Management of patients with atrial fibrillation: different therapeutic options and role of electrophysiology-guided approaches. AB - At present the approach to atrial fibrillation treatment is based on the electrophysiological patterns of atrial fibrillation (on the basis of multiple intra-atrial recordings or sophisticated new mapping techniques) only in a restricted minority of patients, those who are candidate to ablation of the substrate and/or of the triggers. Atrial fibrillation has a broad spectrum of clinical presentations and a heterogeneous electrophysiological pattern. The treatment of this arrhythmia, both with drugs and non pharmacological treatments, has been based, classically, on empirical basis and on a clinically-guided staged approach. The limitations of pharmacological treatment led in recent years to the development of a wide spectrum of non pharmacological treatments. This implies a change in the approach to atrial fibrillation and the need to identify potentially ideal candidates to complex and expensive treatments. In this view it is currently under investigation the possibility to identify potential responders to a definitive treatment or a combination of treatments (both pharmacological and non-pharmacological) on the basis of the electrophysiological pattern. PMID- 11889965 TI - Cytokine levels in patients with a very low left ventricular ejection fraction after open heart surgery. AB - INTRODUCTION: Cardiopulmonary bypass and cardiac operations are obligatorily connected with systemic inflammatory reaction. Production of proinflammatory cytokines is responsible also for negative effects on the myocardial function. OBJECTIVE OF STUDY: Follow-up of the dynamic changes of proinflammatory and antiinflammatory cytokine levels in patients with left ventricular dysfunction during the first week after cardiac surgery. PATIENTS AND METHODS: A total of nine patients with a very low left ventricular ejection fraction (22.75 +/- 0.65%) who had undergone cardiac surgery (for coronary artery bypass grafting or aortic valve reconstruction) were investigated during the first after week operation. The preoperative and postoperative plasma levels of tumor necrosis factor (TNF), interleukin-8 (IL-8) and interleukin-10 (IL-10) were estimated by means of ELISA technique. RESULTS: With respect to the preoperative levels, on the first postoperative day the levels IL-8 (from 9.36 to 16.65 pg/ml) (p < 0.05) and of IL-10 (from 6.93 to 28.09 pg/ml) (p < 0.02) significantly rose with a stepwise decrease down to the seventh day after surgery. From the third to seventh day an insignificant increase in TNF level was also noted. CONCLUSIONS: The results have shown that open heart surgery in patients with a severe left ventricular dysfunction evoked a systemic inflammatory response demonstrated by early increase in proinflammatory cytokine IL-8 and was accompanied by increased level of antiinflammatory cytokine IL-10. Despite stepwise decrease in IL-8 levels, they did not reach the preoperative levels, not even on the seventh postoperative day. (Tab. 1, Fig. 3, Ref. 21.) PMID- 11889964 TI - In vitro effect of stobadine on Fe(2+)-induced oxidative stress in rat liver mitochondria. AB - The authors have studied the susceptibility of two key protective enzymes, glutathione peroxidase (GPX) and glutathione reductase (GR) to the reactive oxygen species (ROS) known to induce oxidative damage in vitro system containing Fe2+/EDTA. The ability of ROS scavanger stobadine to prevent oxidative damage was also studied. Incubation of GPX with Fe2+/EDTA resulted in the significant decrease in its enzyme activity while under the same condition the activity of GR was not changed. The presence of stobadine was effective in protecting GPX from the loss of its activity by in vitro oxidizing agents. The monitoring of the mitochondrial outer membrane dynamics by the method of synchronous fluorescence fingerprint showed that the membrane is involved in these processes. (Fig. 3, Ref. 35.) PMID- 11889966 TI - Cell adhesion molecules in the neural development and plasticity. AB - Cell adhesion molecules (CAMs) are cell surface glycoproteins mediating cell-cell and cell-matrix interactions which play a vital role in the embryonic development of the nervous system as well as in the maintenance and nerve regeneration in adults. In the central nervous system (CNS) three main CAMs, such as neural cell adhesion molecule (NCAM), L1 and N-cadherin have been characterized. In addition to their adhesive properties, CAMs are involved in cell migration, growth of axons, nerve pathways formation and synaptogenesis. The binding of CAMs can activate transmembrane-signalling reactions and thereby contribute to the initiation of cellular response in regulation of synaptic plasticity. CAMs play an important role in learning and memory. The role of CAMs in abnormal development and malignancies provide a wide field of research for diagnostic and therapeutic applications. (Ref. 36.) PMID- 11889967 TI - Landau-Kleffner syndrome: a case of a dissociation between spoken and written language. AB - A case of a girl affected by Landau-Kleffner syndrome (acquired aphasia with convulsive disorder) is reported. The girl showed epileptic seizures and EEG abnormalities. At the age of 7 years 10 months they were followed by onset of aphasic symptoms accompanied with behavioural disturbances. By the age of 9 years she developed a severe verbal auditory agnosia and loss of spoken language expression, but was able to use reading and writing to communicate. This unusual dissociation suggests that spoken or written language can be affected selectively. The girl was followed up to the age of 14 years 6 months and her language recovered well by this age. (Fig. 2, Ref. 42.) PMID- 11889968 TI - Management of a shunt malfunction during pregnancy. AB - In some cases of shunt malfunction in pregnant women the symptoms can be so severe and occur so early that not only the mother but also the baby is threatened. In such a situation the close co-operation of a neurosurgeon and a gynaecologist is mandatory. Both conservative and surgical treatment have to be applied to overcome the time until the delivery carries an acceptable risk for the child. In such a case a pre-term delivery by Cesarean section should be recommended. (Fig. 1, Ref. 2.) PMID- 11889969 TI - Relation of left ventricular hypertrophy to cardiovascular complications in diabetic hypertensives. AB - The presence of diabetes mellitus and other risk factors of atherosclerosis, such as obesity, smoking and hyperlipidemia, in hypertensive patients makes the prognosis worse. Authors compared the clinical findings in diabetic hypertensive patients with and without left ventricular hypertrophy, the presence of which was diagnosed and defined by echocardiography. The study is based on the analysis of hospital records of 115 hypertensive patients treated at our department during the period 1998-1999. Left ventricular hypertrophy (LVH) was defined by echocardiography as left ventricular mass index > 134 g/m2 in men and > 110 g/m2 in women. Left ventricular hypertrophy was found in 79 patients (mean age 64.6 ys) but not in 36 patients (mean age 63.3 ys). Both groups were matched as to age and sex, intensity and duration of hypertension and diabetes, obesity, smoking and hyperlipidemia. In LVH-positive patients, there was a statistically significant incidence of heart failure, mitral regurgitation and renal involvement and a more non-significant incidence of left ventricular diastolic dysfunction, myocardial infarction, chronic atrial fibrillation and stroke than in LVH-negative ones. Left ventricular hypertrophy usually complicates the course of hypertension. Authors recommend to investigate the presence of left ventricular hypertrophy in hypertensives as it carries a much more complicated course of the disease. (Tab. 5, Ref. 28.) PMID- 11889970 TI - Non-pharmacological approach to smoking cessation. AB - Literary data on non-pharmacological approach in smoking cessation have been reviewed. The used methods are depending on particular target groups: patients visiting general practitioners, clients of smoking cessation clinics, pregnant women, hospitalised patients, and adolescents. Community based intervention represents a specific approach. The existing data on non-pharmacological smoking cessation show certain differences with a wide range of the reported cessation rates. Important role of primary health care providers in smoking cessation is evident, however, frequently underestimated. Specific situation in Central and Eastern Europe (CEE) requires evidence based data in this field, taking into consideration also psychological, social and cultural aspects. Only guidelines based upon authentic studies carried out in CEE can be of value for long-term tobacco control programmes. (Ref. 70.) PMID- 11889971 TI - Pharmacoeconomy of adult asthmatics. AB - Asthma bronchiale represents serious social and economic problems in all over the world. Financial expenses are elevated every year. THE AIM: Of the presented study was to analyze the current therapeutic approach in the management of bronchial asthma and to evaluate pharmacoeconomic aspects of treatment in selected outdoor asthmatic patients in region of Kosice (Eastern Slovakia). PATIENTS: The data of the total number 297 patients (183 females--61.6%, 114 males 38.4%), aged from 18 to 78 (average age 38) were analyzed. RESULTS: The total expenses per day for all asthmatics represented 19,465 Sk (65.54 Sk/day/patient). Short-acting beta 2-agonists (beta-A) (55.2%) were administered most frequently, followed by inhaled corticosteroids (CS) (43.1%), theophyllins (T) with slow released formulas (36.4%), cromoglycates (C) (33%), long-acting beta-A (19.9%), combined preparations beta-A with anticholinergics (ATCH) (Berodual, Combivent) (12.12%), depot CS (10.44%), antileucotriens (ALT) (10.10%), combined preparation beta-A with C (Ditec, 8.75%), short-acting T (6.73%), systemic CS (5.72%), ATCH (4.71%). Additive therapy represented antihistamines (AH) (69.36%), topic AH, nasal and eyes drops (38.8%), specific immunotherapy (36.03%), immunomodulatory therapy (23.23%), expectorans and antitusive drugs (15.49%), respectively. C (20.33%) represented the highest financial cost from the total financial budget followed by inhaled CS (16.34%), long-acting beta-A (13.20%) and ALT (8.9%), short-acting T (0.39%) and short acting beta-A (3.41%), respectively. In the group of additive drugs AH dominated (15.85%). CONCLUSION: The optimal selection of antiasthmatic drugs should be kept in mind by physicians. Our study shows some reserves in respect of optimal selection of antiasthmatic (generic drugs should be preferred), therapeutic efficacy and compliance of patients. (Tab. 11, Ref. 15.) PMID- 11889972 TI - [Stability of myocardial perfusion during revascularization under propofol anesthesia]. AB - During coronary artery bypass grafting surgery (CABGS) there are many factors which may disturb myocardial oxygen balance with consequent irreversible myocardial damage. Anesthesiologists are expected to use drugs with tendency to optimize hemodynamics and myocardial oxygen balance, making in that way conditions for preserving myocardial perfusion. In this study, performed on 19 patients undergoing CABGS under dominant propofol anesthesia, there were no signs of intraoperative ischemia manifested by statistically significant dislocation of ST segment in EKG leads D11, V5 and aVL. PMID- 11889973 TI - [Complications in the use of vascularized fibular grafts: classification and treatment]. AB - The complications in the reconstruction of the diaphysed defects by the use of VFG or VOSKFG are not shown in the literature coherent so it is very difficult to compare and use showed results. For the compensation of the bone defects the specific complications could be differentiated according to the influence on the primary and overall efficiency of the reconstruction. The classification of the complications in the donors region and osteogenic and vascular complications in the recipients region was suggested in the analysis which encompassed all these divisions. PMID- 11889974 TI - [Assisted circulation II--systems for intermediate and long-term mechanical circulatory support]. AB - The concept of artificial circulatory support has been established almost 200 years ago. It has only been within the last four decades that physicians and engineers have developed mechanical assist devices that can temporarily support the circulation until the native heart recovers from a reversible injury. In patients who do not regain native heart function, long-term circulatory support or permanent replace (biologically--heart transplant or permanent mechanical circulatory support) is indicated. In this paper we describe the devices (ABIOMED BSV 5000, Thoratec, HeartMate, Novacor and CardioWest TAH), that are in current clinical use for intermediate and long-term mechanical circulatory support. PMID- 11889975 TI - [Fixation of the vagina and neovagina with the sacrospinal ligament]. AB - In order to fix the prolapse of vaginal vault after vaginal or abdominal hysterectomy we use the technique of transvaginal sacrospinous colpopexy. The authors used this technique for the firs time when they have made neovagina at male transsexual. The technique is as follows: after making the tunnel between the bladder and recrum, under the control of the left hand fingers the vagina, or neovagina is fixed to sacrospinous ligament. No complications or recidive followed the operation ever. The authors point out that the skillfull surgeon should know very well the anatomy of both male or female pelvis in order to perform this operation. PMID- 11889976 TI - [Residual choledocholithiasis in elective cholecystectomy and choledocholithotomy]. AB - Cholecystectomy is one of the most frequent abdominal operation. Common bile duct stone may be found in 6-15% of elective cholecystectomics. One or more stones may be left behind within common bile duct. In a series of 106 consecutive cholecystectomics performed in the First surgical clinic in Belgrade over six months period (from 07.08.1997.-31.12.1997.) operative cholangiography was performed routinely in every single case. Unexpected common bile duct stone was found in 13 patients (12.26%). After removal of stones choledochoscopy was performed in 7 out of 13 patients with common bile duct stone (53.8%). An operative T-tube cholangiography was performed in all 13 cases before closure of the abdomen. Postoperatively, usually from 8th to 10th day a secondary T-tube cholangiography in the X-ray department was performed in all 13 cases. Retained stone was found in 2 patients, both in group of patients in whom choledochoscopy had not been carried out. The retained stone was successfully removed endoscopically in one case but the second patient had to be reoperated as endoscopy was unsuccessful. We conclude that choledochoscopy reduces the risk of retained common bile duct stone and it should be practiced whenever possible. PMID- 11889977 TI - Early angiogenic capabilities of the transposed omental flap after omentomyelopexy. AB - Abundant in blood and lymph vessels, capable to adhere to the surface of every lesion, with capillary overgrowing in 4-6 hours, omentum represents almost very suitable organ for revascularization of the ishaemic nervous tissue. Angiographic study of the omentum, especially of the surgically developed omental flap have been rarely performed in clinical practice. The aim of this study was evaluate the angiographic features of the omental flaps after omentomyelopexy. Omentomyelopexy based on the left gastroepiploic vessels was performed in 100 patients of different levels spinal cord injuries. In order to study the omental flap's vascularization, selective angiography of the splenic artery was performed in three patients at the 10th postoperative day. In one patient, angiographic finding showed the establishment of anastomosis between omental flap's arteries and vertebral and spinal artery and in that way, almost incredible angiogenic capabilities of omentum have been proved. Besides confirmation of vitality and good vascularization of the extraperitonealy transposed omental flap, an extraordinary angiogenic capabilities of the omentum have been proved as early as at the 10th postoperative day. To our best knowledge the angiographic findings of this kind have not been published so far. PMID- 11889978 TI - Leiomyosarcoma of the duodenum. A successful reoperation after recurrency. AB - Leiomyosarcomas of the duodenum are rare tumours. Approximately 160 cases have been reported so far. Pain, bleeding and duodenal obstruction causing vomiting and weight loss are the leading symptoms. The mass may be palpable. If resected, prognosis is favorable in a majority of cases, much better then in carcinomas of the duodenum. Patients have to be followed up as eventual recurrencies are frequently suitable for resection. We present a 62 years old male who was submitted to cephalic duodenopancreatectomy for duodenal leiomyosarcoma (reported earlier) who developed recurrency 6 years after original surgery and in whom a successful reoperation was performed. Two years after second surgery the patient stayed disease free. PMID- 11889979 TI - [Surgical importance of anatomic features of the pancreatic head and tail]. AB - The surgical anatomy of the left pancreatic portion includes topography of this entity in relation to the peritoneum and the adjacent organs, variations of the arterial vascularization and venous drainage, and of the ductal system. A particular emphasis was on the practical significance of the variations of the pancreatic tail, the arterial anastomoses of the corporocaudeal region, and the position and morphology of the pancreatic veins. Ending remarks include a small review on distal pancreatectomy. PMID- 11889980 TI - [Wound treatment in ancient Greece]. AB - Wounds probably represented one of the first challenges the first human beings had to face with. Unlike some primitive creatures that are capable to repair even the entire lost part of the body, a simple healing with connective tissue is all that can be expected in human. From the early days through entire history it was tried to find the way to alleviate and to speed up the wound healing and to prevent dangerous or even deadly complications that may take place. A concept set up by Old Greeks that suppuration of wounds is good and laudable maintained almost twenty three hundred years. Through history it was challenged only occasionally, but stayed firm to the end of 19. century when was proved to be wrong. PMID- 11889981 TI - [Esophagus preserving surgery--eversion and mucosal stripping]. AB - The eversional stripping and esophageal mucosal stripping methods as standard procedures in the preserving esophageal surgery are presented in this paper. These procedures have indication limit and results are excellent in selected risk dependent group of patients. This review describes technique, indications and advantages of these techniques in comparision with conventional finger esophageal dissection. PMID- 11889982 TI - [Protoscolecides in the surgical treatment of hepatic echinococcosis--myth or reality]. AB - Traditionally protoscolicidal agents have been used during conservative surgery for cystic liver hydatidosis to sterilize the cyst cavity and to avoid dissemination. Many surgeons questions the effect of the procedure due to lack of objective data and possible complications for their use. Recently introduced minimally invasive techniques--percutaneous (PAIR), endoscopic and laparoscopic- have reestablished the need for the evaluation of protoscolocidal agents. PMID- 11889983 TI - [Significance of C-reactive protein in the treatment of acute necrotizing pancreatitis]. PMID- 11889984 TI - Late histological changes in the transposed omental flap. PMID- 11889985 TI - [Protective colostomy or ileostomy?] ]. AB - A group of patients is presented, where colorectal or coloanal anastomosis after resection was performed together with concomitant covering colostomy (102) or ileostomy (25), during the period 1985-2000. Covering colostomy was most often performed upon hepatic flexure (82.1%) and upon sigmoid colon (8.9%). The incidence of complications during performing or closure of colostomy was 6.8% with one case (0.8%) of death due to unsuccessful protection of primary anastomosis. The incidence of complications in covering ileostomy was 16%, with 3 (12%) deaths. Authors are analysing the data from literature about indications and technique of covering enterostomies, comparing them with their experience and concerning the choice of appropriate protective procedure. They came to conclusion that the reason for covering procedure has been in many instances reasonable, and that in the question of protection both methods are comparable, but covering ileostomy has potentially more dangerous complications. Therefore as a method of anastomotic protection authors are generally recommending covering colostomy except in technically specific cases. PMID- 11889986 TI - [Pelvic exenteration]. AB - Pelvic exenteresis (total, anterior and posterior) is operative procedure reserved for the local advanced malignancies of the pelvis. In the period 1995 2001, we have treated 54 patients (20 male, 34 female) by this procedures. By anterior pelvic exenteresis we have treated 6 females for: Ca vesicae urinary (4 pts). Ca PVU after irradiation therapy (1 patient), Ca urethrae (1 patient). By posterior pelvic exenteresis we have treated 2 females for primary advanced Ca of the rectum. By total pelvic exenteresis we have treated 46 pts (20 male, 26 female): Ca PVU after irradiation therapy (10 females), recidivant Ca PVU (8 females), primary advanced Ca of the rectum (7 male, 1 female), recidivant Ca of the rectum (10 male, 7 female), recidivant Schwanoma (1 male), recidivant Sa stromae endometrii (1 female), recidivant Ca vesicae urinary (1 female). The median survival time of all 54 patients was 24 months. Early postoperative mortality was 18% (10 pts). Twenty patients died with median survival of 18 months (range 4-48 months). Twenty one patients are alive without evidence of disease with median follow up period of 41 months (range 6-60 months). Three patients were lost from follow up. Exenteresis pelvis is very complicate operative procedure and it should be limited to perform only in couple surgical centers. PMID- 11889987 TI - [Evaluation of enterogastric reflux in relation to functional status of the gallbladder]. AB - The aim of the study was estimation of the relation between the gallbladder (GB) motility function and the presence and quantity of enterogastric reflux (EGR). We investigated 172 patients with: physiological GB function (filling and emptying)(FGB), impaired GB function (prolonged filling and ejection fraction < 45%) and afunctional gallbladder (AGB)(without visualization). The study was performed during 90 min (1 f/min) after i.v. application of 185 MB 99mTc-dietil IDA. After 30 min. test meal was given while at the end stomach was marked. According to the parameters from time activity curves over stomach and hepatobiliary system, the index of ERG was calculated, while GB filling and ejection fraction were estimated from the GB time/activity curve. We can conclude that EGR occurs more frequently in the patients with afunctional GB in comparison to those with functional and decreased motor function. Also, EGR quantity is in correlation with the impairment of the GB function. PMID- 11889988 TI - [Mediastinitis--diagnosis and therapy]. AB - Mediastinitis is serious, life-threating infection. Desceding necrotizing mediastinitis is a form of mediastinitis with mortality rate of 40%. It develops as a complication of odontogenic and deep cervical infection. Early diagnosis and therapy are the most important issues. Computed tomographic scan is suggested if odontogenic infection is complicated with dyspnea. Mediastinitis is usually polymicrobial infection. Transcervical mediastinal drainage and intravenous antibiotic therapy are suggested. In some [figure: see text] cases aggressive mediastinal drainage by thoracotomy approach is performed. PMID- 11889989 TI - [Semi-exenteration of the orbit in therapy of malignant conjunctival tumors]. AB - Authors present their experience in treatment of the conjunctival maliganncies by semiexenteration of the orbit. They discuss the advantages and the disadvantages of this procedure. They suggest the use of semiexenteration in stead of exenteration whenever it is possible. PMID- 11889990 TI - [New surgical approaches in splenic diseases]. AB - To improve the success of postoperative results, in the surgeries of the spleen diseases new procedures had been developed from hematological pretreatment to further operative and postoperative treatment of the patient. Elective splenectomy is quite a big anesthetic challenge. In the period from 1996 to 1998 in the Institute for digestive diseases 147 splenestomies had been done as a consequence of the spleen diseases. Before the operation there is no reason for compensate the platelets if their number is equal or bigger than 50 x 10(9)/l unless the patient is bleeding. If the patient has less than 50 x 10(9)/l the operation shouldn't postponed if we have previously prepared enough doses of platelets from the separator the pool, or the ordinary platelets. Now days at our clinic we operate the patients with extremly plateleptema. The operations have been very successful. The team work of surgeons, anestehsiologists and hemathologist is the most important thing. PMID- 11889991 TI - [Acute transfusion-related lung injury]. AB - Transfusion Related Acute Lung Injury (TRALI) is the most serious, potentially lethal, transfusion reaction caused by anti leukocyte antibodies carried passively into recipient's circulation by transfused haemoproduct. It is manifested most frequently within first four hours after transfusion of allogenous haemoproduct containing anti leukocyte antibodies. It is characterized with symptoms and signs of acute respiratory distress syndrome. This rare, but severe transfusion reaction with mortality rate of around 10% is often misdiagnosed, since TRALI signs are assigned to other clinical conditions. Thus, an education for timely recognition and urgent care of TRALI should be initiated. PMID- 11889992 TI - [Ambrose Pare (1510-1590)]. PMID- 11889993 TI - [War injuries of blood vessels of the extremities]. PMID- 11889994 TI - [Resection procedures in liver tumors--intraoperative dilemmas and complications]. AB - Liver resection surgery for tumors has many dilemas and possible intraoperative complications that surgeon has to deal with during his work. Objectives of this work was to, according to our experience, and pointing to a certain anatomical and technically essential moments, makes possible easier decission making concerning the indications, with safer work. Also, we wanted to suggest how to deal with possible complications during the operations. PMID- 11889996 TI - [Laparoscopic resection of the rectum]. AB - Improving the laparoscopic equipments and the techniques have spreaded the repertoire of the laparoscopic procedures. In the beginning, it was performed the laparoscopic-assisted resection of the rectum, and later was performed the entire laparoscopic procedures on the colon. It had presented a patient, 71 years old man with diagnosis of the adenocarcinoma recti. After preoperately prepare carried out the laparoscopic procedure with T-T anastomosis by EEA stapler. We presented the first case of the laparoscopical performed resection of the rectum with desire to show possibilities and advantages of the laparoscopic surgery in the relation with conventional surgery and with hope that this techniques would be found the application in our land. PMID- 11889995 TI - [Cholecystitis associated with milk of calcium] ]. AB - Milk of calcium cholecystitis is a rare disease predominantely of adults. The term designates a pathologic accumulation of calcium carbonate in the gallbladder, much rarer in the common bile duct as well. The patient may be symptom-free. If present the symptoms may be very mild or as an acute biliary pain, transient jaundice and mild attack of pancreatitis. Calcium in the bile may be liquid or it may form a paste-like mass. Both may be seen on a plain X-ray of the abdomen. The stone or stones of calcium carbonate may be formed so that they may cause an obstruction of the gallbladder outflow tract or cystic duct or pass into the common bile duct or into the duodenum. [figure: see text] We present a 71 year old male who was admitted for inguinal hernia repair but in whom a mild hiperbilirubinaemia was found and then on ultrasonography a gallbladder stone was diagnosed. We performed a hernia repair as well as cholecystectomy and operative cholangiography which turn to be normal. Within an almost normal gallbladder a gray-whitesh paste-like mass was found composed of calcium carbonate. The postoperative recovery was uneventful and serum bilirubin become normal. Histology of gallbladder showed chronic nonspecific cholecystitis. PMID- 11889997 TI - Laparoscopic assisted radical right hemicolectomy for cure of cancer. PMID- 11889998 TI - [Topographic and structural characteristics of the minor duodenal papilla]. AB - The study of minor duodenal papilla topography and structure was carried out on 36 fresh autopsy specimens of human duodenopancreas. We performed precise measurements of its distance to the major duodenal papilla and to the superior duodenal flexure. There was no correlation between the position of the minor papilla and the incidence of duodenal ulcer disease. Microdissection and histological staining of the minor papilla did not reveal an anatomically defined sphincter around the terminal portion of the accessory pancreatic duct. All the specimens of the minor papilla contained within acini of pancreatic tissue. A terminal dilation of the accessory pancreatic duct was found in 22% of the cases. PMID- 11889999 TI - The importance of reassessment to chronic wound care. PMID- 11890000 TI - Frequency of peristomal complications. AB - Practicing wound ostomy continence (WOC) nurses see peristomal complications in their ostomy patient population. The exact frequency and cause of these complications are unknown. Research into problems arising from prolonged use of ostomy appliances is lacking. To ascertain the frequency of peristomal complications, two WOC nurses at a major medical center prospectively assessed the peristomal skin of all new ostomy patients returning for their 2-month postoperative check-up using a peristomal complications tool. The study was conducted from August 1999 to August 2000. Descriptive statistics were used to summarize the data. In the course of the study, 161 new ostomy patients were seen, 10 with peristomal complications, for a frequency of 6%. The frequency of peristomal complications was the highest in people with ileal conduits (five of 34, 15%) and ileostomies (four of 46, 9%). Only one of the 81 people with a colostomy developed peristomal complications. All 10 patients had a retracted stoma. Eight had chemical damage (six with irritant dermatitis, two with pseudoverrucous lesions) and two had Candida infections. Although the study sample size is small, the results suggest that patients with ileal conduits and patients with retracted stomas may require more frequent follow-up visits to monitor skin conditions and pouching modifications. PMID- 11890001 TI - Common bacterial skin infections. AB - Skin infections account for a significant portion of dermatologic disease, often resulting in or as a consequence of a disruption in the skin's integrity. This article covers the presentation, diagnosis, and treatment of the more common bacterial infections. The infections presented herein include impetigo, ecthyma, folliculitis, carbuncles/furuncles, cellulitis, toxic shock syndrome, and ecthyma gangrenosum. Once a diagnosis is made, treatment is based on the culture and antibiotic sensitivities of the offending organisms. PMID- 11890002 TI - Cost-effectiveness of Apligraf in the treatment of venous leg ulcers. AB - Venous ulcers are the most common chronic wounds of the lower leg. Skin substitutes recently have been introduced to stimulate nonhealing wounds. To conduct an incremental cost-effectiveness analysis, a model was developed to compare the four-layer bandage system, with and without one application of skin substitute, for the outpatient treatment of venous leg ulcers. The model estimated the costs and consequences of treatment with and without the skin substitute application. Two analytic horizons were explored: 3 months and 6 months. Determined by seven physicians, data and assumptions for the 3-month model were based on information from a clinical trial, published studies, and clinical experience. Data for the 6-month model were extrapolated from the shorter model. The model results indicate that over 3 months, the use of the skin substitute provided a benefit of 22 ulcer days averted per patient at an incremental cost of $304 (societal). The incremental cost-effectiveness ratio was $14 per ulcer day averted. Over 6 months, the incremental cost-effectiveness ratio was less than $5 per ulcer-day averted. The skin substitute plus a four layer bandage was more costly and more effective than the four-layer bandage alone. The skin substitute is increasingly cost-effective over a longer analytic horizon and in a subgroup of patients with ulcers of long duration (greater than 1-year duration at baseline). The results come from a model that is based on a series of estimates and assumptions, and accordingly, confirmation of this finding in a prospective study is encouraged. PMID- 11890003 TI - Pressure ulcers, outcomes, and the nursing shortage. AB - The incidence of pressure ulcers is dependent on a number of variables. This article argues that the current and upcoming nursing shortage will adversely influence the pressure ulcer-related goals of public policy initiatives that strive to reduce the humanistic and cost burden of certain chronic conditions. The current state of the nursing shortage is described herein, along with examples of legislation proposed to control for this crisis and a consideration of the just distribution of goods and services (e.g., pressure ulcer care). Also provided is the voice of a nurse profoundly affected by the situation--a consideration of real and perceived threats to care. PMID- 11890004 TI - No release, no refund. PMID- 11890005 TI - When the patient you're treating has been injured in an accident. PMID- 11890007 TI - It's time to add knowledge, skills and maturity to the dental education mix. PMID- 11890006 TI - Somatoform salivary complaints. Case reports. AB - Patients with salivary gland complaints are seen with a large array of signs and symptoms. Usually these patients have an underlying pathophysiological process that can account for their symptoms. However, in a significant number of patients, no known biological process can be found that would account for the patient's complaint. In such cases, somatization is a possible cause. Somatization is a frequently cited feature of patients with various forms of mental illness. In this paper, we will attempt to illustrate the classic signs of a somatoform disorder in three different patients whose diverse salivary complaints fulfill the criteria for a diagnosis of somatoform disease. PMID- 11890009 TI - NYS health department increases Medicaid dental fee schedule. PMID- 11890008 TI - Oral health status of Asian and Hispanic women. A pilot study. AB - Data on the oral health of Asian and Hispanic women are rare in the dental literature. The objective of this study was to investigate the oral health status and needs of women from these two groups. Forty newly registered women from each group were the subjects of the study. Standard forms were used to collect sociodemographic information, medical histories, oral health status, oral hygiene practices and smoking habits. The oral examination recorded the DMFT and periodontal condition. Data were analyzed using SPSS Univariate General Linear Model, odds ratios and relative risks. Chi-Square tests were within the 95% confidence interval, and with @ = .05. The results showed that there was no statistical difference in age, education, oral health care, smoking habits, annual dental visits or DMFT between the two groups. PMID- 11890010 TI - When the patient is hearing-impaired. PMID- 11890011 TI - Respectfully disagree. PMID- 11890012 TI - We're in the Army now. PMID- 11890013 TI - Oral Health America's National Oral Health Report Card. PMID- 11890014 TI - Se Habla ... La Paz affords a unique educational opportunity for dental assistants. PMID- 11890015 TI - Painful, cracked, dry skin of the hands. An occupational hazard for some, made worse in winter. PMID- 11890016 TI - Professional liability coverage provided by your ADAA membership. PMID- 11890017 TI - Materials for restoration of primary teeth: 2. Glass ionomer derivatives and compomers. AB - A wide variety of materials have been used for the restoration of primary teeth. Resin-modified glass ionomers and the more recently introduced viscous glass ionomers have been developed from conventional glass ionomer materials in an attempt to overcome the suboptimal properties of conventional glass ionomers. These materials would appear to have the necessary physical characteristics for restorations in load-bearing situations in primary teeth, as have the resin-based compomer materials, which now have well documented success rates in a number of studies. The first of these two articles described 'traditional' restorative materials, including amalgam and conventional glass ionomer, for the restoration of primary teeth. This paper describes materials derived from traditional glass ionomers in an attempt to overcome the suboptimal properties of conventional glass ionomers and resin-based materials such as compomer. PMID- 11890018 TI - Digital records in orthodontics. AB - In this electronic age there is a general move towards keeping digital records, and many trades and professions now use digital images exclusively. In this article, the advantages (and occasional disadvantages) of the use of digital photography, digital radiography and the latest development--digital study models -in orthodontics are discussed. PMID- 11890020 TI - Clinical governance: something new for general dental practice? AB - This article is intended to inform general dental practitioners about the background to clinical governance, how it came about and how the government has introduced it into the NHS. Because clinical governance is new in general dental practice, how it eventually becomes incorporated is still debatable, but this article attempts to describe ways in which practitioners can use it to their advantage. It explains how a cultural change is needed in dental practice to embrace these issues of quality and that issues of clinical governance have become a General Dental Service 'Terms of Service' requirement for all practitioners. PMID- 11890019 TI - Dental ceramics: what's new? AB - The advances in dental ceramic materials and systems continue to be related to improvements in strength, fitting accuracy and aimed towards avoidance of the use of metal substructures both in posterior and anterior teeth. Many of the changes seen within the last few years have been associated with modifications to, and improvements of, existing techniques. These are considered in this paper, and ceramic post systems are also reviewed. PMID- 11890021 TI - The management of traumatically intruded permanent incisors in children. AB - Trauma to the permanent dentition, particularly the maxillary incisors, is common. Prompt and appropriate management can significantly improve the prognosis for many of these dentoalveolar injuries. Unfortunately, much of this trauma is left untreated. This paper discusses the management of children who present with intruded permanent incisors. PMID- 11890022 TI - Proud to be a dinosaur? PMID- 11890023 TI - The new MFGDP(UK). PMID- 11890024 TI - Exarticulation and reimplantation utilizing guided tissue regeneration: a case report. AB - The extraction and reimplantation of teeth is a technique that has been practiced for many years, resulting in successful retention of teeth for up to 30 years. Although evidence confirming the advantages of this technique is limited, clinical observations and histologic examinations have revealed some important factors that are prerequisites for success: limitation of the time the tooth is exposed to the extraoral environment and preservation of vital periodontal tissue attachments on root surfaces. The latter goal can be achieved by gentle, atraumatic removal of the tooth from its socket, and hence the term exarticulation and reimplantation is more representative of the technique. In the present case report, a maxillary lateral incisor with a developmental defect was treated by exarticulation and reimplantation and application of calcium sulfate. Exarticulation and reimplantation seems to be a useful clinical procedure, but controlled studies are required to confirm its efficacy. PMID- 11890025 TI - Oral piercing and gingival recession: review of the literature and a case report. AB - A young adult developed severe gingival recessions and radiographic signs of trauma to the periodontium after wearing a tongue barbell and a lip stud. Oral body art (piercing) can be hazardous to the periodontium; nevertheless, patients inclined to such practices do not see them as health hazards and are very reluctant to remove them. PMID- 11890026 TI - Pulp-dentin biology in restorative dentistry. Part 7: The exposed pulp. AB - Exposure of the dental pulp, through a caries lesion, accidentally during routine cavity preparation, or as a result of tooth fracture, is a clinical reality that requires optimal treatment. The potential for healing by formation of a dentinal bridge is good, provided that the pulp is not inflamed. Calcium hydroxide has a long history of inducing dentinal bridge formation to promote successful healing. Resin composites are emerging as alternative materials for pulp capping, but healing is slower, and relatively little clinical experience is available for analysis. The prognosis for healing is poor after exposure of an inflamed pulp. Pulpotomy should be considered to create a wound in an uninflamed location. If an exposure is expected through a caries lesion, stepwise excavation of the carious dentin should be considered to allow healing of the dentin and pulp prior to the final excavation of the carious dentin. PMID- 11890027 TI - Microleakage of new all-in-one adhesive systems on dentinal and enamel margins. AB - OBJECTIVE: The aim of this study was to compare the microleakage of new all-in one adhesive systems on enamel and dentin margins with that of a conventional total-etch system. METHOD AND MATERIALS: Thirty buccal Class V cavities were prepared with enamel and dentin margins in 30 bovine teeth and randomly divided into three groups of 10 specimens each. Group 1 was bonded with Etch & Prime, group 2 with Prompt L-Pop, and group 3 with 35% phosphoric acid plus Prime & Bond 2.1. After being stored in an environment of 100% humidity for 24 hours, the teeth were immersed in a 50% silver nitrate solution for 24 hours and then put in a developing solution for 15 minutes. The specimens were sectioned vertically and buccolingually, and microleakage was evaluated on a scale of 0 to 3. RESULTS: For the enamel margin, statistical analysis revealed that there was no significant difference between groups 1 and 2 or groups 1 and 3 but that there was a statistically significant difference between groups 2 and 3. Statistical analysis revealed no significant difference among the groups in dentinal microleakage. CONCLUSION: Among the three adhesive systems used in this study, Prompt L-Pop provided the least microleakage in enamel; however, there was no statistically significant difference among the groups on dentin margins. PMID- 11890028 TI - A comparison of strengths of five core and post-and-core systems. AB - OBJECTIVE: In this in vitro study, the strength of several core and post-and-core systems was compared. A second aim was to compare the strength of posts and cores on root-filled teeth with that of cores on vital teeth. The failure modes of the core and post-and-core systems were also studied. METHOD AND MATERIALS: For root filled teeth, Composipost carbon fiber dowels and gold alloy posts and cores were tested. For vital teeth, glass-ionomer cement with threaded parapulpal retention pins, resin composite with threaded parapulpal retention pins, and gold alloy with parallel parapulpal pins were tested. Specimens were tested in a Zwick universal material-testing machine. RESULTS: A significant variation in strength was found among core systems of cast gold, resin composite, and glass-ionomer cement constructed on vital teeth. The strength of the systems constructed on root-filled teeth did not vary significantly. The mode of failure varied, depending on the core or post-and-core material. CONCLUSION: Composipost posts and cores and cast gold posts and cores were equivalent in strength and did not vary significantly from gold cores constructed on vital teeth. The low strength values obtained for glass-ionomer cement in combination with threaded retention pins makes this combination a poor choice for core buildup. PMID- 11890030 TI - Dental lights: where has the simplicity gone? PMID- 11890029 TI - A clinical, microbiologic, and radiographic study of deep caries lesions after incomplete caries removal. AB - OBJECTIVE: Clinical, radiographic, and microbiologic changes in deep caries lesions were assessed after incomplete carious dentin removal and tooth sealing. METHOD AND MATERIALS: Thirty-two teeth with deep caries lesions were studied. Treatment consisted of incomplete excavation of the demineralized dentin, application of calcium hydroxide, and sealing for a 6- to 7-month period. The color and consistency of the dentin were clinically assessed. Differences in radiographic density were assessed by digital image subtraction. Microbiologic samples were obtained from the demineralized dentin before the temporary sealing and after the experimental period. The samples were cultivated on blood agar under aerobic and anaerobic conditions, in Mitis Salivarius agar, and Rogosa selective Lactobacillus agar. RESULTS: Two cases were lost during the study; one presented pulpal necrosis. In the other case, there was pulpal exposure during removal of provisional sealing. In all teeth, the initial demineralized dentin was soft and wet; one lesion was yellow, 21 were light brown, and eight were dark brown. After treatment, the dentin was dry, and 80.00% of specimens were hard, 16.67% were leathery, and 3.33% remained soft. The dentin was light brown in five teeth and dark brown in 25. There was a statistically significant mean difference in radiographic density (pixel intensity), 88.77 +/- 7.02 in the control areas and 94.66 +/- 6.75 in the test areas. The counts of anaerobic and aerobic bacteria, lactobacilli, and Streptococci mutans had decreased significantly by the end of treatment. CONCLUSION: Incomplete removal of carious dentin and subsequent tooth sealing resulted in the arrest of the lesions, suggesting that complete dentinal caries lesion removal is not essential to the control of caries lesions. PMID- 11890031 TI - Oral verruca vulgaris. PMID- 11890032 TI - The functional and esthetic deficit replaced with an acrylic resin gingival veneer. AB - Periodontal attachment loss in the maxillary anterior region can often lead to esthetic and functional clinical problems including disproportional and elongated clinical crowns, visible interdental embrasures, and altered linguoalveolar labiodental consonant production. Assuming fixed prosthetic reconstructions will be chosen to treat these areas, it becomes a hygienic compromise to fill these areas in with porcelain. In the presence of these problems, an acrylic resin gingival veneer is an easily constructed, inexpensive, and practical device to optimize the esthetic and functional outcome in these special situations while permitting cleansibility of the prosthesis and supporting tissues. This article presents a step-by-step technique for the fabrication of a gingival veneer. PMID- 11890033 TI - Comparison of luting cements for minimally retentive crown preparations. AB - OBJECTIVE: The purpose of this study was to compare the retentive strengths of resin, glass-ionomer, and zinc phosphate cements under adverse conditions. METHOD AND MATERIALS: Thirty extracted teeth were mounted and prepared in their long axis. The axial wall height was 3 mm and the convergence angle was 28 degrees. These conditions increased the role of the cement and decreased the role of the preparation in providing retention of the casting. The axial surface area was determined. Copings were fabricated with a ring aligned in the long axis to facilitate removal of the crown. They were cemented with a resin cement, a glass ionomer cement, or a zinc phosphate cement. A block randomization scheme was used to assign cements so that the mean surface areas of the teeth were equivalent in all groups. The copings were loaded in tension, and the amount of force required to remove the coping was recorded. The stress required to remove the coping was calculated. RESULTS: The mean stress required to remove the copings was 9.4, 5.0, and 3.1 MPa for the resin, glass-ionomer, and zinc phosphate cements, respectively. CONCLUSION: The resin cement group was significantly stronger than both the glass-ionomer cement and the zinc phosphate cement groups. The glass ionomer cement was significantly stronger than the zinc phosphate cement. PMID- 11890034 TI - "BMT, a challenge in nursing management". PMID- 11890035 TI - [In the interests of taking one's time even when one reads a scientific article! Critical review of the literature on tooth eruption]. AB - Nowadays everybody has access to the medical literature, not only through the libraries of the universities, but even more conveniently via Medline and Pubmed. It is very tempting only to read abstracts and summaries instead of "wasting time" by reading the whole article. Reading the article itself however enables the reader to select only these articles that have a good methodological and statistical basis. In this contribution several topics will be discussed that may help the reader to read scientific articles with a critical mind: definitions, quality of the data, study concepts, composition of the sample, control group, statistical analysis, sample size and interpretation of the results. Different items are exemplified with articles that deal with the emergence of permanent teeth. PMID- 11890036 TI - [Analysis of dental caries risk factors in the primary dentition]. AB - The aim of the present study was to assess indicators shown to be associated with the prevalence of caries in the primary dentition of 7-year-old Flemish schoolchildren. Cross-sectional first year data of the Signal-Tandmobiel survey were analysed (n = 4468). Gender, age, oral hygiene habits, use of fluorides, dietary habits, geographical factors and social factors were involved in logistic regression models. There was a significant difference (P < 0.05) in caries experience determined by the geographical spread, with an explicit trend of caries declining from the east to the west. A start of brushing before the age of 2 and a brushing frequency of at least once a day need to be encouraged, while the use of sugar containing drinks and snacks between meals needs to be restricted to a maximum of 2 per day. The mean dmf-t values were the lowest for the most advantaged children and where threefold higher in the least advantaged children. There was a cumulative effect of decreasing social level and oral health habits on the caries prevalence. Social inequalities in oral health among children exist in Flanders. PMID- 11890037 TI - [Factors determining dental plaque and their relation to the assessment of caries among 7-year-old Flemish children]. AB - The present investigation aimed to assess the impact of a number of variables on the variation on the amount of dental plaque present in the mixed dentition of 7 year-old Flemish children and to investigate a potential correlation between caries experience and the amount of dental plaque. Analyses were performed with ordinal plaque index as outcome measure, considering the children's school (i.e. class) as a random effect. This random effect turned out to be of great importance when dealing with large clustered samples. From the multiple logistic regression analysis, it was concluded that the older the subjects the more dental plaque was observed (or = 1.35; CI: 1.08-1.68; P = 0.0081). Four exploratory variables, could be identified as important risk indicators for dental plaque accumulation in the mixed dentition: 'start of brushing after the age of 2 years' (or = 1.161; CI: 1.09-1.23; P < 0.0001), 'brushing frequency of < 1 x/day (or = 1.37; CI: 1.12-1.66; P = 0.0019), more than two in between meal snacks (or = 1.18; CI: 1.03-1.36; P = 0.02) and daily intake of sugar containing drinks (or = 1.24; CI: 1.08-1.42; P = 0.002). Further, being male was linked to a higher amount of plaque (or = 1.16; CI: 1.00-1.34; P = 0.0506). Finally a weak but significant correlation (spearman's rho: r = 0.23; P < 0.0001) between caries experience and the amount of cervical plaque was shown. PMID- 11890038 TI - [Oral hygiene and gingival condition among 12-year-old children in the Brussels Region]. AB - The study analyses oral hygiene and gingival status in a group of 12-yr-old children from the Region of Brussels. In 1998, a total of 496 children from eight selected schools participated in the sample. All children were interviewed about their socio-economic status and oral health care. Records of the plaque index and the gingival index were made. The mean plaque and gingivitis were 1.24 (+/- 0.03) and 1.32 (+/- 0.03), respectively. Seventy percent of the examined sites presented plaque and gingivitis. Privileged children showed lower means than non privileged counterparts. Multiple linear regression showed that dental plaque was significantly associated to age, toothbrushing and appointment in case of discomfort or pain (P = 0.02). Age, gender, type of the toothbrush and use of dental floss were associated to gingivitis (P = 0.05). Daily home-based mechanical plaque removal is critical for the maintenance of gingival health and when efficiently performed it leads to remission of gingivitis. Dentists should be encouraged to give information and training on regular plaque removal to their patients. PMID- 11890040 TI - The precision removable restoration: attachments in prosthodontics. PMID- 11890039 TI - [Fluorosis: diagnosis, risk assessment and epidemiology]. AB - Fluorosis is the most widespread side-effect of fluoride use and appears as discrete white spots on the enamel up to severe enamel dysplasia. There are different techniques for scoring fluorosis in epidemiological surveys. In the literature there is no uniform way of selection of teeth and data processing. Fluorosis risk is determined by environmental factors such as water and food fluoride content as well as individual factors such as use (or misuse) of fluoride supplements and fluoridated oral hygiene products. In a group of Flemish schoolchildren, fluorosis prevalence is low and mainly related to use of fluoride supplements and toothpaste in childhood. PMID- 11890041 TI - Dentistry and the future of professional ethics. PMID- 11890042 TI - Human genome, oral infection/systemic diseases & the future of clinical dentistry. PMID- 11890043 TI - Thoughts on setting up an ethics program. PMID- 11890044 TI - Occupational hand injuries in the dental profession. PMID- 11890045 TI - Iron content in musculus longissimus lumborum et thoracis (m.l.l.t.) of fattening pigs. AB - Pork is among the most essential dietary sources of iron in Germany, considering its beneficial absorption rate. During 1997 and 1998 meat samples from 52 pigs per year were collected at the Thuringian performance testing station in order to examine the total iron content of pork. Musculus longissismus lumborum et thoracis (13./14. thoracic vertebra) from the right side of the carcasses were prepared for analyses. The total iron content was measured by atomic emission spectroscopy with inductively coupled plasma. The iron content of the meat was 4.18 +/- 0.97 mg/kg fresh material. This result was significantly lower than corresponding data from literature. An influence of the genetic composition of the pigs examined on the total iron content could be observed. The daily gain (R = -0.26) and intramuscular fat content (R = -0.28) were inversely correlated to the iron content. However, the lean meat proportion (R = 0.26) as well as haem pigment content (R = 0.27) were directly correlated to the total iron content of the meat. PMID- 11890046 TI - Antioxidant activity of sweetgrass (Hierochloe odorata Wahlnb.) extract in lard and rapeseed oil emulsions. AB - The antioxidant activities of sweetgrass (Hierochloe odorata) and sage (Salvia officinalis) extracts were studied in emulsions of lard and rapeseed oil using soy lecithin as an emulsifier, and addition of cupric acetate as an oxidation catalyst. The antioxidant activity was about the same in the two substrates. The stability against autoxidation was substantially increased by both sweetgrass and sage extracts and their combination. The stability was particularly high, if citric acid and/or ascorbyl palmitate were added to plant extracts. PMID- 11890047 TI - Capacity of edible seaweeds to modify in vitro starch digestibility of wheat bread. AB - The capacity of two species of edible seaweeds (Wakame, Undaria pinnatifida and Chondrus, Chondrus crispus) to modify the rate of white bread starch digestibility by an in vitro digestion system, as well as glucose retardation index and apparent viscosity were studied. Both Algae showed different effect on glucose retardation index. While Wakame did not make difficult dialysis of glucose, it only retained 2.50 +/- 2.99% to respect a negative control. Chondrus impaired the diffusion of a 28.70 +/- 2.35% of glucose, which are very close to those of citrus pectin (31.50 +/- 4.12%). On the other hand, viscosity of Chondrus solution was higher than Wakame and slightly lower than citrus pectin solution. The analysis showed a very low content of total starch in Wakame (0.51%) and Chondrus (0.47%). Algae reduced the digestion of white bread starch. The profile of starch hydrolysis was characteristic for each alga. Glycaemic index estimated from the degree of starch hydrolysis within 90 min was low (79) with respect to white bread (value 100). Seaweeds showed a suitable capacity to modify in vitro starch digestibility of white bread. Chondrus produces a more pronounced response. This fact seem to be due to different composition of samples. PMID- 11890048 TI - Effect of metal ions on structure and activity of papain from Carica papaya. AB - Papain, a powerful proteolytic enzyme, is an endoprotease belonging to cysteine endopeptidase family. It is used extensively in food processing especially in tenderization of meat. In this study, we have made an attempt to show the structure activity relationship of this enzyme and the role of calcium and magnesium ions in the activity and stability of the enzyme. Results of activation and stabilization of the enzyme by these cations showed concentration dependent effect. The enzymatic activity of papain increases to a maximum of 18% and 24% in presence of calcium and magnesium ions at 1 x 10(-3) M concentration, respectively. Thermal denaturation studies showed that the binding of calcium and magnesium ions bring about change in the thermal stability of papain at various concentrations of these metal ions. Far ultraviolet circular dichroic studies showed no significant change in the alpha-helix and beta-sheet structure of the papain upon binding of these metal ions. The mechanism underlying the structure activity relationship of papain in presence of these metal ions have been discussed here with reference to the ionic radii, ligand binding preference, coordination numbers and the electrostatic forces between the protein molecule and cations present in the microenvironment of the enzyme. PMID- 11890049 TI - An application of linear regression technique for predicting bulgur yield and quality of wheat cultivars. AB - Grains of 26 Turkish wheat cultivars and advanced breeding lines were used in this study. Simple correlations between a number of quality parameters to predict bulgur yield and bulgur cooking quality were determined. Highly significant correlations between bulgur yield and each of the thousand-kernel weight and the sum of the grain over 2.8 + 2.5 mm sieves were obtained for both durum and bread wheat samples (p < 0.01). The regression equations showed that the models involving two variables (the thousand-kernel weight and the thickness of the grain for durum wheat samples; the thousand-kernel weight and the length of the grain for bread wheat samples) resulted in the highest R2 values. For an assessment of the influence of all factors on bulgur cooking properties (total organic matter: TOM and colorimetric test values), simple and multiple regression analyses were used to find equations that predict best the relationship between various quality parameters and bulgur cooking properties. The models involving two variables; the vitreousness and the dry gluten contents for the durum wheat samples and SDS sedimentation test value and wheat protein content for the bread wheat samples resulted in the highest R2 for the TOM value. PMID- 11890050 TI - Commercial milling of suni bug (Eurygaster spp.) damaged wheat. AB - In some wheat growing countries, considerable quantities of commercial wheat are rendered unusable in standard milling because of pre-harvest damage of the grain by protease-injecting insects. The possibility of mitigating the detrimental effects of bug damage by eliminating the mill streams of lower quality from straight run flour was investigated. The changes in the amount of damaged kernels by cleaning and washing prior to milling were also examined. Dry cleaning decreased the level of insect damage from 26.4% to 23.0%. A further improvement to 12.8% was obtained by removing the light-density bug-damaged kernels by washing. There were substantial differences among the mill streams of the commercial mill as evaluated by the standard and modified sedimentation tests and standard alveograph test. The best streams were those from the first, second, third and fourth reductions. PMID- 11890051 TI - Heat-resistance characteristics of ascospores of Eurotium chevalieri isolated from apricot juice. AB - A heat-resistant fungus was isolated from aseptically packaged apricot pulp. The fungus was identified as Eurotium chevalieri. Heat resistance of the fungus was studied at four different temperatures (70, 75, 80 and 83 degrees C) after activation of its ascospores for 30 min at 70 degrees C. D70, D75, D80 and D83 values of ascospores of Eurotium chevalieri were estimated by linear regression (log-survival vs. heating time) as 118.58, 34.15, 5.50 and 3.77 min, respectively. The z-value was determined in the same way (regression of log-D values vs. heating temp.) and was found as 8.23 degrees C. PMID- 11890052 TI - Effect of activated charcoal on patulin, fumaric acid and some other properties of apple juice. AB - In this study, 0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 g/l amounts of activated charcoal (AC) were added into apple juice with a patulin content of 62.3 ppb obtained from a well-established manufacturing company. Apple juice samples were then mixed for 0, 5, 10, 20, and 30 min, respectively. Considerable reduction in the patulin and HMF values was found while there is a dramatic improvement in the colour and clearness of apple juice. However, AC did not cause a significant decrease in the fumaric acid level of apple juice. The best result was obtained at 3.0 g/l AC mixed for 5 min. In addition, a negligible reduction in brix and pH values of samples was observed. PMID- 11890053 TI - Monitoring of pesticide residues on cucumber, tomatoes and strawberries in Gaza Governorates, Palestine. AB - Three techniques of gas chromatography (GC) either with flame photometric detector (FPD), electron capture detector (ECD), or with mass-spectrometry (MS) were applied for identification and quantification of pesticide residues on 45 samples of cucumber, tomatoes, and strawberries in fifteen locations in Gaza Governorates. GC-FPD analysis showed the presence of four different organo phosphorus (OP) pesticides, their levels were very low and below maximum residue limits (MRL's). GC-ECD detected ten different pesticides at levels below the MRL's. Using the GC-MS technique, alpha and beta-endosulfan, chlorpyrifos, carbofuran, chlorfluazuron, triadimenol I and II, penconazole, coptafolmetabolite, pyrimethanil and iprodione were detected and confirmed on some samples of cucumber, tomatoes and strawberries. All GC-MS pesticide residues detected on tomato were below the MRL's except chlorfluazuron while on strawberry were below the MRL's except penconazole, chlorfluazuron and pyrimethanil, but on cucumber were slightly higher than the MRL's except alpha and beta-endosulfan. Also, statistical analysis of pesticide residues in all samples showed that most of the detected residues mean were significantly lower than the MRL's (p < 0.05). Generally, tomatoes showed the least number and level of pesticide residues by all the GC-techniques. On the other hand, strawberries showed greater number and levels of pesticide residues, particularly by the GC-MS technique. These results indicate that the protective period to elapse before harvesting should be increased especially on strawberry. The results also can help in risk assessment of consumers exposure to the expected pesticide residues. PMID- 11890054 TI - Canned cod liver as a source of n-3 polyunsaturated fatty acids, with a reference to contamination. AB - Lipid composition (HPLC), fatty acid composition (GC/MS), lipid oxidation (peroxide value, anisidine value), UV-VIS and fluorescence spectra (Ex 365 nm), and susceptibility of lipids to oxidation (photooxidation test) as well as heavy metal, PCB, and DDT contents were determined in canned, raw, and thermally treated cod liver (separately in the released oil and in the solids). Canned products of three manufacturers were examined. Mean contents of n-3 polyunsaturated fatty acids (n-3 PUFAs) in the oil and solids were 31.91 +/- 1.83 and 16.59 +/- 7.48 g/100 g, respectively, the respective contents of docosahexaenoic acid (DHA) being 17.88 +/- 1.69 and 8.79 +/- 3.67 g/100 g. Lipid resistance to oxidation was found to decrease after thermal treatment of livers. However, the lipid oxidation level in canned liver stored for 3-8 months was not high and averaged, for the entire can content, 0.47 +/- 0.4 Meq O, the oil being more susceptible to oxidation that the solids. It is concluded that canned cod liver is a very good source of n-3 PUFA, particularly with respect to DHA. Heavy metal, DDT, and PCB contamination and the presence of lipid oxidation products in the canned products tested remain at a level producing no perceivable health hazard and could in no way interfere with consumption of recommended amounts of n 3 PUFAs. PMID- 11890055 TI - Determination of total cholesterol content in food by flow injection analysis with immobilized cholesterol oxidase enzyme reactor. AB - A new analytical method has been developed--including a new sample preparation procedure--for the automatic determination of total (free and bound) cholesterol in food by flow injection analysis (FIA) with immobilized (cholesterol oxidase) enzyme reactor (IMMER). A suitable sample preparation procedure has been applied to eliminate the problems derived from sensitivity of FIA equipment to common organic solvents used for dissolving of cholesterol: after direct saponification the non-saponificable fraction was dissolved in water phase detergent (sodium cholate) solution. The analytical method is based on the oxidation by cholesterol oxidase followed by the photometric determination of hydrogen peroxide (at 500 nm) using the indicator reaction with peroxidase. The new FIA method was tested for commercial food samples such as whole egg powder and dried pasta. The results were compared with data obtained by GC-determination. It was found that this new procedure is suitable for rapid automated measurement of total cholesterol content in foodstuff and consequently, the FIA technique with immobilized enzyme reactor could be an alternative to the widely used gas-chromatographic (GC) method. PMID- 11890056 TI - Bacteriological characteristics of dressed young pigeon (squabs) Columba livia domesticus. AB - A total of 50 frozen squabs carcasses were collected from different retail markets in Cairo and Giza governorates. The collected samples were examined bacteriologically. The aerobic plate count, the enterobacteriaceae and the Staphylococcus count were 6.6 x 10(5), 6.3 x 10(2) and 1.4 x 10(3) cfu./gram, respectively. The results revealed no positive samples for Salmonella Yersinia entercolitica, Listeria monocytogenes and Campylobacter jejuni. Staphylococcus aureus was isolated from 2% of samples. The sources of contamination of processed squabs carcasses and measures to minimize the bacterial load and safe guard the consumer are mentioned. PMID- 11890057 TI - Effects of season on vitamins A and E contents and colour of Turkish butter. PMID- 11890058 TI - Fe, Ca and Mg contents in selected fast food products in Poland. AB - The Fe and Mg contents in selected fast food products available in restaurants and fast food outlets in Poland were determined by AAS, and the Ca content by AES. The mean Fe contents in the studied fast food products were from 0.7 to 2.3 mg/100 g, or from 0.6 to 2.3 per single serving. The highest means for this element were found in a serving of hamburger (2.3 mg), fishburger (2.0 mg) and chicken sandwich (2.0 mg). The mean Ca contents in the studied products were from 11.6 to 192.2 mg/100 g, or 10 to 192.2 mg per serving. The highest means for this element were found in a serving of pizza (192.2 mg) and cheeseburger (134.8 mg). The mean Mg contents in the studied products were from 6.8 to 34.1 mg/100 g1 or 5.9 to 37.3 mg per serving. The highest means for this element were found in a serving of french fries (37.3 mg), chicken sandwich (34.7 mg) and fishburger (30.4 mg). Based on the Fe, Ca and Mg contents found in these products, the percentage of the Recommended Dietary Allowance (RDA) of these elements was calculated for one serving of each product. These calculations were done for various groups of people in Poland. The highest percentage of the recommended Fe intake could be covered by one serving of hamburger (15-23% RDA), fishburger (14 20% RDA), or chicken sandwich (13-20% RDA). The highest percentage of the recommended Ca intake could be covered by one serving of pizza (17-24% RDA) or cheeseburger (12-17% RDA); and for Mg one serving of french fries (11-19% RDA), chicken sandwich (10-17% RDA), or fishburger (9-15% RDA). From the conducted studies it may be concluded that some fast food products can serve as a source of Fe, Ca and Mg in the diet of people of various ages. PMID- 11890059 TI - Value of add-ons to eradicate errors adds up. PMID- 11890060 TI - Growth factors and their potential use in bone grafting procedures for dental implants. PMID- 11890061 TI - Endothelin-1 expression in the microvasculature of normal and 3-hour continuously loaded rat molar periodontal ligament. AB - Endothelin-1 (ET-1) distribution is described in the microvascular bed (MVB) of normal rat molar periodontal ligament (PDL). Five male Sprague-Dawley rats, aged 90 days, were anaesthetized and an external pressure of 100 +/- 20 g, maintained for 3 hours, was transmitted occlusally to randomly allocated left or right molars. The controls were the contralateral molars. Rats were perfused for 5 minutes with 5 per cent paraformaldehyde, and the mandibles post-fixed and stored in 30 per cent sucrose. Sagittal, undemineralized, mandibular jaw sections, approximately 150 microns thick, were immunolabelled with ET-1 and alpha-smooth muscle actin (alpha-SMA) primary antibodies, and IgG/CY5 and IgG/CY3 secondary antibodies, respectively. Serial images were captured with a Bio-Rad MRC-1000 UV confocal laser scanning microscope (CLSM). In the control PDL, the ET-1 immunolabelling occurred sporadically in all categories of PDL blood vessels. ET 1 showed a punctate distribution within endothelial cells, producing longitudinal and circumferential pan-endothelial labelling. Immunoreactivity to the ET-1 antibody occurred at or adjacent to vessel branching sites, and affected blood vessels with and lacking alpha-SMA immunolabelling. Treatment effects on ET-1 immunofluorescence were analysed for vascular endothelium, socket bone surface cells, cementum surface cells and PDL background in the cervical, inter-radicular and apical regions. Significant (P < 0.05) region by treatment interactions occurred for the endothelium and bone. Cementum showed a significant (P < 0.05) region effect and a significant (P < 0.05) treatment effect. However, the region by treatment interaction was not significant. Background immunofluorescence showed significant (P < 0.05) region by treatment effects for the endothelium and bone. ET-1 activity is the default state for normal PDL vascular endothelium. PMID- 11890062 TI - Mesio-marginal findings at tilted molars. A histological-histomorphometric study. AB - The aim of this study was to investigate the mesio-marginal findings at tilted molars (TM) by means of histological-histomorphometric evaluation. Eight lateral tooth bone segments with TM (six mandibular, two maxillary) from males aged 20-32 years were compared with those of eight samples with non-tilted molars (NTM) in males aged 18-35 years. In comparison with the NTM samples, the TM revealed a higher amount of supra- and subgingival plaque, a significantly higher total number of inflammatory cells (P < 0.05) and blood vessels (P < 0.05) in the connective tissue adjacent to the junctional epithelium, and a lower density and corono-apical width of gingival fibres. No significant differences (P > 0.05) were found between the mesio-marginal bone level of the TM (mean: 978 microns) and that of the NTM (mean: 1222 microns). In contrast, indications were found that TM may affect the disto-marginal bone level of the mesial tooth. PMID- 11890063 TI - Tissue reaction to orthodontic tooth movement--a new paradigm. AB - Direct or indirect resorption are both perceived as a reaction to an applied force. This is in contrast to orthopaedic surgeons who describe apposition as 'the reaction to loading of bone'. The article reviews the literature on intrusion of teeth with periodontal breakdown, and on the basis of clinical and experimental studies. The conclusion is reached that intrusion can lead to an improved attachment level, and that forces have to be to low and continuous. The tissue reaction to a force system generating translation of premolars and molars in the five Macaca fascicularis monkeys is described. Three force levels, 100, 200, and 300 cN were applied for a period of 11 weeks. Undecalcified serial sections were cut parallel to the occlusal plane and a grid consisting of three concentric outlines of the root intersected by six radii was placed on each section so that areas anticipated to be subject to differing stress/strain distributions were isolated. A posteriori tests were utilized in order to separate areas that differed with regard to parameters reflecting bone turnover. Based on these results and a finite element model simulating the loading, a new hypothesis regarding tissue reaction to change in the stress strain distribution generated by orthodontic forces is suggested. The direct resorption could be perceived as a result of lowering of the normal strain from the functioning periodontal ligament (PDL) and as such as a start of remodelling, in the bone biological sense of the word. Indirect remodelling could be perceived as sterile inflammation attempting to remove ischaemic bone under the hyalinized tissue. At a distance from the alveolus, dense woven bone was observed as a sign of a regional acceleratory phenomena (RAP). The results of the intrusion could, according to the new hypothesis, be perceived as bending of the alveolar wall produced by the pull from Sharpey's fibres. PMID- 11890064 TI - The temporomandibular joint and the disc-condyle relationship after functional orthopaedic treatment: a magnetic resonance imaging study. AB - Causative correction of Class II skeletal malocclusions may be achieved through bite jumping by various means. Numerous animal experiments have yielded evidence of remodelled temporomandibular structures after mandibular protrusion. However, the mode and extent of structural and/or topographic changes of the disc-condyle relationship after functional orthopaedic treatment is still unresolved. A problem exists in defining the physiological position of the condyles and disc condyle relationship, which is tentatively determined by various methods particularly in magnetic resonance tomographic studies. Despite the high resolution provided, the results have to be interpreted with caution, as osseous resorption and apposition cannot be assessed by visual evidence. This investigation examined the impact on the temporomandibular joints (TMJ), i.e. the condylar shape and position, and the disc-condyle relationship, of the bionator plus extra-oral traction in combination with vertical elastics. The underlying reactions were studied by means of magnetic resonance images (MRI) obtained from n = 15 successfully treated patients (mean age 11.6 years). PMID- 11890065 TI - Hyoid bone position after surgical mandibular advancement. AB - A cephalometric evaluation of the changes in the horizontal and vertical placement of the hyoid bone and of those in the position of the head over the cervical spine after surgical mandibular advancement was undertaken. Seven linear and one angular measurement were investigated in 60 patients, 17 males and 43 females, before and one year after surgical mandibular advancement. The hyoid bone moved forwards horizontally in 78 per cent of the subjects, backwards in 17 per cent, and in 5 per cent of patients it retained its pre-operative position. It moved vertically downwards in 32 per cent of subjects, and in 63 per cent it moved upwards and closer to the body of mandible. The amount of horizontal and vertical change of the hyoid bone was associated with the corresponding change of the mandible after surgery. The vertical change was more distinct in females compared with males. There was variation in the position of the head over the cervical spine; it showed extension in 26.7 per cent of the sample, flexion in 71.7 per cent, and remained the same in only 1.6 per cent after surgery. The results show that with surgical mandibular advancement the hyoid bone follows mainly the advancement of the mandible and moves closer to the body of the mandible. However, there are variations in the changes of hyoid bone and head position that are difficult to predict. PMID- 11890066 TI - Maxillary morphology in obstructive sleep apnoea syndrome. AB - The aim of this case-control study was to test the hypothesis that maxillary morphology differs between obstructive sleep apnoea (OSA) patients and non snoring, non-apnoeic subjects. Forty randomly selected patients [36 M, 4 F; mean age 49 +/- 2 (SEM) years] with varying degrees of OSA (mean Apnoea/Hypopnoea Index 32 +/- 4/hour) were compared with 21 non-snoring, non-apnoeic control subjects (18 M, 3 F; mean age 40 +/- 2 years). An intra-oral assessment of the occlusion was carried out, particularly for the presence or absence of posterior transverse discrepancies. Maxillary dental arch width was assessed by standardized lateral inter-tooth measurements (inter-canine, inter-premolar, and inter-molar) from dental models. Palatal height and maxillary depth were also measured. The maxillary dental arch was described by a 4th order polynomial equation. The ratios of maxillary to mandibular width (max/mand) and maxillary to facial width (max/facial) were determined from standardized postero-anterior cephalometric radiographs in a subgroup of patients (n = 29) and all controls. Twenty patients (50 per cent) had evidence of posterior transverse discrepancies compared with one control subject (5 per cent; P < 0.01). All patients had significantly reduced inter-canine, inter-premolar, and inter-molar distances (P < 0.05). The maxillary depth was also shorter (P < 0.05), but palatal height was not different. The quadratic coefficient of the polynomial equation was greater in the patients than in the controls (P < 0.05), indicative of greater arch tapering. Patients had smaller maxillary to mandibular and maxillary to facial width ratios (P < 0.01). These results suggest that OSA patients have narrower, more tapered, and shorter maxillary arches than non-snoring, non-apnoeic controls. Further work is required to determine the relevance of these findings in the pathophysiology of OSA. PMID- 11890067 TI - Orthodontic aspects of the use of oral implants in adolescents: a 10-year follow up study. AB - The aim of the present study was to evaluate the long-term effect of implants installed in different dental areas in adolescents. The sample consisted of 18 subjects with missing teeth (congenital absence or trauma). The patients were of different chronological ages (between 13 and 17 years) and of different skeletal maturation. In all subjects, the existing permanent teeth were fully erupted. In 15 patients, 29 single implants (using the Branemark technique) were installed to replace premolars, canines, and upper incisors. In three patients with extensive aplasia, 18 implants were placed in various regions. The patients were followed during a 10-year period, the first four years annually and then every second year. Photographs, study casts, peri-apical radiographs, lateral cephalograms, and body height measurements were recorded at each control. The results show that dental implants are a good treatment option for replacing missing teeth in adolescents, provided that the subject's dental and skeletal development is complete. However, different problems are related to the premolar and the incisor regions, which have to be considered in the total treatment planning. Disadvantages may be related to the upper incisor region, especially for lateral incisors, due to slight continuous eruption of adjacent teeth and craniofacial changes post-adolescence. Periodontal problems may arise, with marginal bone loss around the adjacent teeth and bone loss buccally to the implants. The shorter the distance between the implant and the adjacent teeth, the larger the reduction of marginal bone level. Before placement of the implant sufficient space must be gained in the implant area, and the adjacent teeth uprighted and paralleled, even in the apical area, using non-intrusive movements. In the premolar area, excess space is needed, not only in the mesio-distal, but above all in the bucco-lingual direction. Thus, an infraoccluded lower deciduous molar should be extracted shortly before placement of the implant to avoid reduction of the bucco-lingual bone volume. Oral rehabilitation with implant-supported prosthetic constructions seems to be a good alternative in adolescents with extensive aplasia, provided that craniofacial growth has ceased or is almost complete. PMID- 11890068 TI - Growth hormone treatment promotes guided bone regeneration in rat calvarial defects. AB - This study evaluated the biomechanical strength and bone formation in calvarial critical size bone defects covered with expanded polytetrafluoroethylene (e-PTFE) membranes in rats treated systemically with recombinant human growth hormone (rhGH). A full-thickness bone defect, 5 mm in diameter, was trephined in the central part of each parietal bone in 40 one-year-old female Wistar rats, which were randomly assigned to two groups of 20 animals each. The bone defects were covered with an exocranial and an endocranial e-PTFE membrane. From the day of operation, the rhGH-treated animals were given 2.7 mg rhGH/kg/day and the placebo injected rats were given isotonic sodium chloride. The animals were killed 28 days after operation. The biomechanical test was performed by a punch out test procedure placing a 3.5-mm diameter steel punch in the centre of the right healed defect. After mechanical testing, the newly formed tissue inside the defect was removed and the dry and ash weights were measured. The left healed defects were used for three-dimensional (3D) reconstruction by means of micro-computer tomography (micro-CT). Ultimate load, ultimate stiffness, and energy absorption at ultimate load were significantly increased in the rhGH-treated group (P < 0.003). Also, tissue dry and ash weights were significantly augmented in the rhGH treated group (P < 0.001). The 3D reconstruction of newly formed bone showed that there was almost twice as much bone volume present in the rhGH-treated defects compared with the placebo group. The experiment demonstrated that rhGH administration enhances bone deposition and mechanical strength of healing rat calvarial defects, covered with e-PTFE membranes. PMID- 11890070 TI - Are pre-treatment psychological characteristics influenced by pre-surgical orthodontics? AB - A number of investigations have looked at psychological changes occurring in association with orthognathic treatment. However, most of these studies have used a pre-surgery questionnaire as the baseline measurement. There is little data relating to the true baseline, i.e. that prior to any active treatment. Until this aspect is investigated, it is not possible to assume that pre-surgery is an acceptable baseline. This questionnaire based study aimed to assess changes in six psychological outcome measures between T1 (prior to any active treatment) and T2 (following pre-surgical orthodontics/prior to surgery). The outcome variables were: state anxiety, trait anxiety, depression, self-esteem, body image, and facial body image. Sixty-two patients (39 females and 23 males) completed both questionnaires. The results showed that intervention, in the form of orthodontic treatment, had a minimal effect on the chosen psychometric outcome variables. There was a significant reduction in satisfaction with body image amongst patients who initially reported mild to moderate dental/facial problems, whilst a moderate increase in satisfaction occurred in those patients reporting severe conditions initially. Also of note were significant increases in state anxiety amongst older patients whilst trait anxiety showed greater increases in females than males. PMID- 11890069 TI - Bite force in pre-orthodontic children with unilateral crossbite. AB - In the present study bite force was examined in pre-orthodontic children with unilateral posterior crossbite and compared with an age- and sex-matched control group. The sample comprised 52 children aged 7-13 years, 26 pre-orthodontic children with unilateral posterior crossbite (crossbite group), and 26 children with neutral occlusion (control group). Unilateral bite force was measured at the first molar by means of a pressure transducer. Furthermore, symptoms and signs of temporomandibular disorders (TMD) and number of teeth in contact in the intercuspal position (ICP) were recorded. In both groups, the maximum bite force increased significantly with age and with increasing stages of dental eruption, but the bite force in both sexes did not differ significantly. There were no significant differences in bite force between sides, but this was significantly smaller in the crossbite group than in the controls (P < 0.001). Regression analysis showed that stage of dental eruption (P < 0.001), number of teeth in occlusal contact (P < 0.01), and unilateral crossbite (P < 0.001) were the only variables significantly correlated with bite force. The number of teeth in contact was significantly smaller in the crossbite group than in the controls (P < 0.05) and the frequency of muscle tenderness was significantly higher in the crossbite group than in the controls (P < 0.05). These results suggest that differences in the muscle function associated with unilateral crossbite lead to a significantly smaller bite force in the crossbite group compared with controls and this difference did not diminish with age and development. These findings indicate that early treatment of unilateral posterior crossbite is advisable to optimize conditions for function. PMID- 11890071 TI - A resin veneer for enamel protection during orthodontic treatment. AB - The aims of this study were to test the tensile bond strength of a recently developed veneer. Sound premolar teeth (120) extracted for orthodontic purposes were divided into two experimental and two control groups. In one experimental group (V1) 4-META/MMA-TBB resin (4META) was used on the surface veneer prepared with micro particle filled resin (MFR) as an adhesive for bracketing and in the second group (V2) 4META was applied on the surface veneer with the trial resin. For the controls, in group R 4META was used on the enamel surface without veneer and in group G light-cured glass ionomer cement was applied. The 30 samples in each group were divided into three groups of 10 samples and thermal cycled (TC) at 3000, 10,000 or left uncycled. Tensile testing was carried out using an Instron machine. After tensile testing the bond failures in the experimental groups were recorded using a stereomicroscope. Statistical analysis was performed using ANOVA. In group V2 the resin veneer was able to maintain sufficient bond force to enamel during clinical use. The bond strength of group V1 was significantly higher than that of groups R (P < 0.05) and G (P < 0.01) at TC 0, but for both TC 3000 and 10,000, the bond strength of group V1 was lower than groups R and G, respectively. There were significant differences between groups V1 and R (P < 0.01) for TC 3000, and between groups V1 and R and G (P < 0.01) at TC 10,000. The bond strength of group V2 was almost equal to that of group R at TC 0. At TC 3000, group V2 showed significantly lower bond strength than group R (P < 0.05), but no significant difference was found compared with group G. At TC 10,000, there were no significant differences between groups V2, R or G. When comparing groups V1 and V2, the bond strength of group V1 was significantly higher than that of group V2 (P < 0.01) at TC 0, but the bond strength of group V1 was significantly lower than that of group V2 for both TC 3000 (P < 0.05) and TC 10,000 (P < 0.01). Comparison between groups R and G, showed that the bond strength of group R was significantly higher than that of group G for both TC 0 (P < 0.01) and TC 3000 (P < 0.01), but no significant difference was found for TC 10,000. In group V2, nine samples showed adhesive failure between the veneer surface and bracket adhesive before thermal cycling. There were significant differences between the MFR and both trial resin and glass ionomer cement (P < 0.01) when examining thermal expansion. No significant difference was found between the trial resin and glass ionomer cement. It is suggested that application of a resin veneer prior to bracket bonding is suitable for clinical application to protect the teeth and to prevent decalcification and caries. PMID- 11890072 TI - Applying split-thickness skin grafts: a step-by-step clinical guide and nursing implications. AB - Wounds in the lower extremities represent a complex medical dilemma and a significant financial burden on the healthcare system. Often, skin grafts and flaps must be incorporated into the treatment of complex defects. Wound management has developed from a multidisciplinary to a collaborative forum, blurring treatment lines among the specialties. Various clinicians and specialists are playing an increasingly larger role in the healing algorithm of wounds in the lower extremities, including the application of skin grafts. This allows the plastic surgeon to deal with the more complex free flaps and microscopic surgical procedures. A patient's status following skin grafting can be an issue in many nursing care environments. This paper provides a pictorial review of a reliable split-thickness skin graft technique that fosters imbibition and inosculation. The points relevant to nursing care include nursing implications (for the graft and donor site), complications, and what to expect in patients who have undergone the procedure. PMID- 11890073 TI - Prevalence and incidence studies of pressure ulcers in two long-term care facilities in Canada. AB - A study was initiated to determine the prevalence and incidence of pressure ulcers in two long-term care facilities in Canada, one with 95 residents and the other with 92 residents. The prevalence study was conducted at both facilities on a single day. The incidence study was completed after 41 and 42 days, respectively, at each facility. Data were collected on demographics, medical information, and possible contributing factors. Each resident was assessed for the presence of a pressure ulcer. Each ulcer was staged and anatomical location was noted. The prevalence of pressure ulcers in the two long-term care facilities was 36.8% and 53.2%, respectively. The incidence of pressure ulcers in the two long-term care facilities was 11.7% and 11.6%, respectively. In conclusion, the pressure ulcer prevalence is higher than published figures for the long-term care setting. However, a pressure ulcer incidence of less than 12% in each facility suggests an equal and acceptable level of nursing care in both facilities. The disparity of pressure ulcer prevalence between the two facilities may be explained by a difference of case mix. PMID- 11890074 TI - A historical overview of pressure ulcer literature of the past 35 years. AB - This article discusses a study of publications on pressure ulcers from 1965 to 1999 using the information available in Medline. Results show that .06% of all articles relate to pressure ulcers. Of all the articles about pressure ulcers, 49% were research articles and 51% were clinical articles. When comparing the total percentage of articles on pressure ulcers to the costs of pressure ulcers in healthcare (1.3% of the total Netherland healthcare budget), one can conclude that clinicians and scientists insufficiently appreciate pressure ulcers as a problem. The number and the proportion of pressure ulcer articles are growing, as well as initiatives to urge researchers studying the same topic to start international working groups, but more research is needed. This article offers insight into the current status of pressure ulcer literature and addresses some reasons for the lack of interest in the pressure ulcer problem. PMID- 11890075 TI - The impact of musculoskeletal changes on the dynamics of the calf muscle pump. AB - Many articles have been published on assessing and treating chronic venous insufficiency and venous leg ulcers; most recommend correcting the underlying cause. These same articles often fail to examine and address a common factor or cofactor of venous hypertension--musculoskeletal changes. Frequently, these changes accompany major injuries, neurological disease, vascular insufficiency, debilitating diseases, myositis, and bone and joint pain and can adversely affect the dynamics of the calf muscle pump. The calf muscles rapidly waste and weaken with disuse--even a change in gait related to a painful ulcer can exacerbate venous hypertension and cause calf muscle disuse atrophy. This article reviews the cause and effect of musculoskeletal changes on the hemodynamics of the calf muscle pump. Recommendations for changes in practice will be based on the identification of the underlying cause of chronic venous insufficiency related to these musculoskeletal changes. PMID- 11890076 TI - Chronic wound caring ... a long journey toward healing. AB - Healthcare professionals use words like "frustrating," "expensive," and "time consuming" to describe chronic wound care. Healing a wound that has been present for an extended period of time is difficult. Often the problem is not just the wound but also the "woundedness" of the individual with the wound. The patient's needs in chronic wound care often continue over months, years, or even a lifetime. This article addresses more than the wound--it offers healthcare professionals' accounts of patient stories and their active involvement in the long journey toward chronic wound healing. PMID- 11890077 TI - Screening evaluation of an ionized nanocrystalline silver dressing in chronic wound care. AB - The successful topical treatment of chronic wounds requires adequate debridement, bacterial balance, and moisture balance. An ionized nanocrystalline silver dressing was evaluated through an uncontrolled, prospective study of a case series of 29 patients with a variety of chronic nonhealing wounds. The four arms of the study included nine patients with foot ulcers, six patients with venous stasis ulcers, two patients with pressure ulcers, and 12 patients with miscellaneous wounds. All wounds were assessed for the usual signs of clinical infection, with most of these parameters being measured and recorded. Microbiologically, bacterial load was determined via quantitative biopsies and semi-quantitative swabs. In general, the results showed a marked clinical improvement for the majority of wounds treated with the dressing. Among improved parameters included decreased exudate and decreased purulence. The quantitative bacterial biopsies did not show any decrease in organism numbers, although the semi-quantitative swabs indicated a decrease in the wound surface bacterial loading. This was indicative of the dressing's ability to reduce surface bacteria and achieve an element of bacterial balance in the superficial dermal compartment. The proposed mechanism of action for this ionized nanocrystalline based dressing is through bacterial and moisture balance within the superficial wound space compartment. PMID- 11890078 TI - High bacterial load in asymptomatic diabetic patients with neurotrophic ulcers retards wound healing after application of Dermagraft. AB - Diabetic neuropathic foot ulcers are a major healthcare burden. These chronic wounds always have a bacterial load, and although normal flora is not harmful, increased tissue burden may impede healing before clinical signs of infection are evident. In this study, chronic noninfected diabetic neuropathic foot ulcers (those with adequate blood supply and pressure offloading) were assessed for bacterial burden immediately before the application of a skin substitute. Eight patients with diabetic neuropathic foot ulcers greater than 1 cm2 and free of necrotic tissue had 3-mm tissue biopsies taken from the ulcer base for quantitative bacteriology. Five of the eight patients (75%) had greater or equal to 10(5) colony forming units/gram organisms present despite the absence of clinical signs of infection. Wound healing rates were linked to bacterial load as determined from quantitative biopsy--no growth was associated with a wound healing rate of 0.2 cm per week, 10(5) to 10(6) colony forming units/gram was associated with a healing rate of 0.15 cm per week, and greater than 10(6) colony forming units/gram was associated with 0.05 cm/per week healing rate. High bacterial burden impeded healing both before and after the application of the skin substitute. The authors will change their clinical practice to assess all diabetic neuropathic foot ulcers using quantitative skin biopsies before applying skin substitutes. All patients will be treated with combination antibiotics and repeat biopsies obtained with decreased bacterial burden (< 10(6) colony forming units/gram) prior to using any bioengineered skin substitute or growth factor treatment. PMID- 11890079 TI - A coordinated approach to chronic wound care. PMID- 11890080 TI - Justice and the right to a decent minimum of healthcare. AB - Throughout time, caring for those who were unable to provide for themselves was based on the virtues of charity, compassion, and benevolence rather than on the ethical principle of justice. In America, and other industrialized countries, the primary barrier to healthcare is the widespread lack of adequate insurance reimbursement. This raises questions about who should receive what share of the nation's scarce healthcare resources, irrespective of insurance status. This article argues that the debate about social justice has been driven by inequalities in access to healthcare and health insurance. This debate is fueled by dramatic increases in healthcare costs. Wound care clinicians are important stakeholders in this debate as they are likely to speak for the patient who has been literally or politically silenced because of age, chronic illness, or poverty. PMID- 11890081 TI - Examining threats to skin integrity. AB - Intact skin serves a vital role in maintaining homeostasis of the body and is regarded as the body's first line of defense against invading micro-organisms. The skin's barrier function can be jeopardized or threatened by several events: aging, dryness, bathing technique, activities of daily living, and soaps. In this paper, the authors review the structures of the skin that facilitate moisture retention and examine bathing practices that threaten the integrity of the skin. Finally, wound and skin care nurses, as well as generalist nurses, are encouraged to critically review the type of soap and the techniques used for bathing. PMID- 11890083 TI - Management and prevention of venous leg ulcers: a literature-guided approach. AB - Managing venous leg ulcers involves management techniques that are indicated both in the treatment of all chronic leg ulcers and those that are specific to venous leg ulcers. The first step in managing venous leg ulcers is performing a holistic assessment of the patient. Once this is complete, any systemic or local factors that may affect wound healing should be addressed. This approach to managing the whole patient is critically important because if significant general wound healing factors are not treated, other specific attempts at healing the venous ulcer will be fruitless. This paper reviews nutritional supplementation, wound bed preparation, antimicrobial therapy, venous insufficiency, compression therapy, different bandage systems, therapeutic adjuncts to compression therapy, and recent advances in vascular surgery. Recurrence prevention also is discussed. PMID- 11890082 TI - Common viral and fungal skin infections. AB - Skin infections account for a significant portion of dermatologic diseases, often resulting in, or as a consequence of a disruption in the skin's integrity. This paper covers the presentation, diagnosis, and treatment of the more common viral and fungal skin infections. The viral infections presented in this paper include herpes simplex virus, herpes zoster, condyloma acuminata, and molluscum contagiosum. The fungal infections presented include tinea pedis, tinea cruris, tinea capitis, tinea unguium, tinea versicolor, and candidiasis. Once a diagnosis is made, treatment with appropriate antifungal, antiviral, destructive, or immune modifying therapies can be instituted. PMID- 11890085 TI - The meaning of trust in healthcare. PMID- 11890084 TI - Pressure ulcer staging. PMID- 11890086 TI - Is diabetic foot care efficacious or cost effective? AB - The incidence of diabetes-related lower extremity amputations continues to increase in the developed and developing world, costing nearly $2 billion and an estimated 2,600 patient-years of hospital stay per year in the United States alone. However, the federal government and private healthcare providers fail to place preventative intervention high on their respective healthcare policy agendas. This manuscript briefly evaluates literature that might support or refute the value of structured, regular care of the diabetic foot and wound. Although the literature is still glaringly sparse, an emerging body of research supports the fact that aggressive, proactive care may result in fewer lower extremity amputations and possibly a higher quality and quantity of life for patients with diabetes. PMID- 11890087 TI - Algorithms, critical pathways, and computer software for wound care: contemporary status and future potential. AB - The contemporary use of algorithms, critical pathways, and computer software significantly affects all aspects of healthcare treatment from primary care to acute trauma management. Guidelines and heuristic devices specific to chronic wound care have been introduced to assist with quality implementation and monitoring of outcomes in a cost-conscious American healthcare system. Some wound care algorithms and critical pathways even have been computerized. This article discusses the differences among the various forms of wound care heuristic methods, describes their advantages and disadvantages, and suggests how they can be successfully implemented in clinical practice settings. Directions for future research also are offered. PMID- 11890088 TI - Reliability of wound measuring techniques in an outpatient wound center. AB - Wound measurements determine whether treatment(s) should be continued or changed. A busy wound clinic must rely on many different personnel for wound measurements. The realization that using a variety of measurement techniques could effect medical treatment choices raised concerns. To determine the inter-rater reliability of wound measuring techniques used by clinical staff in an outpatient wound center, three approaches to wound measurement were studied with the intent to standardize clinic procedures in the authors' facilities and to use the method with greatest inter-rater reliability. An exploratory descriptive study was initiated in a busy multidisciplinary wound-healing clinic in a northeastern Ohio 500-bed teaching and community hospital. Participants included 16 wound care professionals who staff an outpatient wound center. Inter-rater reliability measures were compared to three measurement techniques. The intraclass correlation coefficient was used as the statistical measure of inter-observer agreement. After comparing measurements made by the usual method used by an individual, the clockwise method, and the perpendicular method, the perpendicular method showed clear superiority in agreement among clinicians (ICC .962; df = 11,143; P = < 0.001) as compared to the clock-wise method (ICC .682; df = 11,154; P = < 0.05). This study provided a basis for standardizing the approach to wound measurement among physicians and nursing personnel and has important implications for effective medical care, research, and healthcare cost savings. PMID- 11890089 TI - Wound healing and lymphedema: a new look at an old problem. AB - As the science of wound healing has evolved in the past two decades, so has awareness of the "hidden epidemic" of lymphedema. Substantial information has been accumulated regarding the pathophysiology and therapy of lymphedema. However, until now, the relationship between wound care and lymphedema has received little attention. This review outlines this relationship and asserts that proper wound care demands attention to the related lymphedema. The differences between edema and lymphedema, as well as current evidence-based support defining the negative effect of lymphedema on wound healing, are discussed. The principles of compression therapy in wound related lymphedema also are addressed. PMID- 11890090 TI - How to prepare the wound bed. AB - This clinically focused article addresses the nuts and bolts of wound bed preparation. Preparing the wound bed is a frequently ignored step in the process of treating chronic wounds. In these days of high-tech instrumentation use for chronic wound care (i.e., adjunctive therapies, growth factors, and skin substitutes), clinicians should not forget the basics that are essential for optimizing wound healing. This article introduces the PREPARE Model (see Figure 1) as an aid for guiding care providers in wound bed preparation. PMID- 11890091 TI - Ostomy supplies--out of balance. PMID- 11890092 TI - Cancer profile in Nepal. PMID- 11890093 TI - Existing situation & problems surrounding clinical oncology in Bangladesh. AB - Over all Government health infrastructure for health care delivery in Bangladesh is very good, though current scenario in relation to the Clinical Oncology in Bangladesh administered by a National Council for cancer control is inadequate. And the facilities for creating cancer awareness, screening, diagnosis, and treatment (i.e. multidisciplinary approach, radiation treatment facilities, availability of cancer chemotherapeutic agents, palliative care facilities are not sufficient enough to meet the need of cancer afflicted patients. Most of the patients present themselves in an very advanced stage due to illiteracy, ignorance, lack of cancer awareness, religious prejudice, cheaper and easy availability of non traditional i.e. quackery treatment, inadequate diagnostic facilities to most of the cancer centers, and low socioeconomic status. Top ranking malignancies in male are: Bronchogenic carcinoma, carcinoma of the oropharynx, esophagus, oral cavity, stomach, hypopharynx, carcinoma of unknown primary sites (CUP), lymphoma and carcinoma of the liver. Top ranking malignancies in female are: Carcinoma of the cervix, breast, esophagus, bronchus, oropharynx, ovary, larynx, stomach, oral cavity, & hypopharynx. Irrespective of sex, the top ranking malignancies are: carcinoma of the bronchus, esophagus, oropharynx, cervix, & larynx. Head neck cancers comprise about more than one third of all malignancies, which is more significant findings in our country. Early detection and prevention of cancer deserves a serious thought and urgent attention especially for developing country like ours, where cost benefit analysis is the predominant factor. PMID- 11890094 TI - Genetic polymorphism of estrogen- and carcinogen-metabolizing genes in association with breast cancer risk in Taiwanese women. PMID- 11890095 TI - An overview of novel and therapeutic approaches to hepatocellular carcinoma- Japanese experiences. PMID- 11890096 TI - Molecular lesions as targets for diagnosis and therapy of lung cancer. AB - Alteration of the p53 tumor suppressor gene is one of the most frequently seen molecular lesions in human lung cancer. It is of importance to integrate translational science into clinical practice. In our laboratory, we are in search for clinical utility of p53 gene alterations in management of patients with lung cancer. Here, we would like to discuss p53 alterations as a prognostic factor for lung cancer patients or as a predictor of chemosensitivity. PMID- 11890097 TI - The treatment of advanced non-small-cell lung cancer (NSCLC)--overseas updated and local experiences. PMID- 11890098 TI - Recent advances in the treatment of gynecologic malignancies: an overview of the GOG phase III trials. PMID- 11890099 TI - New strategies and advances in the management of cervical carcinoma. AB - BACKGROUND: Carcinoma of the cervix is still common female cancer in Taiwan. A large portion of patients die of the disease in spite of various kinds of therapies and even the combination of multi-modality treatment. METHODS: The current review includes a comprehensive review of the literature. Many new strategies and advanced in the management of cervical carcinoma have been the subject of this intense study and include assisted systems to improve sensitivity and specificity of Pap smear, human papillomavirus (HPV), and its vaccination, operation, radiation, chemotherapy, and combination of these strategies. Because of the constant failure rate, a systemic and critical review of the disease and management should be considered. RESULTS: With improvement of detection rate of pre-invasive cancer and popularity of screening protocol, the prevalence rate of cervical carcinoma decreases progressively. However, although new strategies in managing cervical carcinoma focus on combination therapy, the life quality and survival rate of these patients with cervical carcinoma only show beneficial in certain group patients. CONCLUSION: As to the complexity of both disease nature and various combinations in the diagnosis and treatment of cervical cancer, comprehensive quality control programs and large prospective multicenter trials are mandatory for establishing the "standard of care" adhered to the principles of evidence-based medicine. PMID- 11890100 TI - Suppression of Helicobacter pylori-induced gastritis by urease inhibitors in Mongolian gerbils. PMID- 11890101 TI - Intervention and follow-up on human esophageal precancerous lesions in Henan, northern China, a high-incidence area for esophageal cancer. AB - BACKGROUND: Esophageal cancer (EC) remains a leading cause of cancer-related deaths in Linzhou (formerly Linxian) and Huixian of Henan province, northern China, which has been well recognized as the highest incidence area for EC. The lack of useful chemoprevention agents and early detection methods is the key factors for stable EC incidence in these areas. Human esophageal carcinogensis has been considered as a multistep progressive process. The natural history for EC, however is not very clear. METHODS: Follow-up studies with linear repeated biopsies and histopathological examination were performed on 778 subjects from Linzhou and Huixian. Of these subjects, 578 subjects were followed for 11 years (1989-2000), 400 subjects with different severity of esophageal precancerous lesions were randomly divided into 2 groups for intervention studies with calcium and decaffeinated green tea (DGT). Each group included 200 subjects (100 subjects for treatment, and 100 subjects for placebo). In calcium group, each subject received an oral supplementation of 1,200 mg of calcium daily for 11 months. In DGT group, each subject received 5 mg of DGT daily for 12 months. In placebo group, each subject received placebo pill for 11 months (calcium group) and 12 month (DGT group). At the entry and the end of the trial, esophageal biopsy specimens were taken at the middle and the lower thirds of the esophagus and from macroscopic lesions, if only, of each subject. RESULTS: DGT trail did not show apparent difference between the treatment and placebo group in alleviating the esophageal precancerous lesions and abnormal cell proliferation. For the calcium intervention study, after 11 years' follow-up, 10 subjects had developed into cancers in the calcium group (10%, 8 EC and 2 GCA), and 8 subjects developed into EC in the placebo group (8%). All these patients were diagnosed at very early stage of cancer (symptom-free). Of the 578 subjects, 25 (18 males and 7 females) had developed into EC (n = 23, 4.3%) and gastric cardia cancer (GCA, n = 2, 0.3%), during the 11 years' follow-up. The mean time of cancer development (from entry of the follow-up study to the cancer detection) was 5.0 +/- 2.9 years (males) and 4.7 +/- 3.2 years (females). Of the 25 patients with EC and GCA, 11 were from the 387 followed subjects with "normal" histomorphology of biopsy at the entry of the follow-up study (3%, 11/387), 2 were from the subjects with basal cell hyperplasia, grade I (BCH I, 2%, 2/94), 7 from the subjects with BCH grade II (BCH II, 10%, 7/72), and 5 from BCH III and dysplasia (20%, 5/25). CONCLUSIONS: DGT trail was not shown to have beneficial effects in alleviating esophageal precancerous lesions and abnormal cell proliferation patterns. Calcium supplementation did not produce apparent long-term effects on EC. BCH II could be considered as precancerous lesions of EC. The quantitative histopathological analysis in terms of number of proliferating basal cell layers is of importance in determining the high-risk subjects for EC and evaluating the intervention results. Follow-up studies with repeated endoscopic biopsies are the powerful strategy for early detection and mortality control of EC and GAC in the high incidence area. PMID- 11890102 TI - Clinical oncology current status in Korea. PMID- 11890103 TI - A review of the natural history of cervical intraepithelial neoplasia. AB - Cervical Intraepithelial Neoplasia (CIN) is a premalignant lesion of cervical squamous cell carcinoma which over time may persist unchanged, regress to normal or a lesser grade of CIN, or progress to a higher grade of CIN or invasive carcinoma. Rates of progression correlate directly with the CIN grade. Human Papilloma Virus (HPV) detection is a significant determinant of CIN regardless of grade. Studies of risk factor profiles, cytogenetic abnormalities, cell proliferation indices, cell cycle and senescence control, oncogene and tumor suppressor gene expression, protein expression, and HPV status have been conducted to identify determinants of CIN I and CIN II/III and predictors of CIN 1 progression. Differences in these attributes suggest that CIN I is a sexually transmitted, productive HPV infection, whereas CIN II/III is a dysplastic lesion resulting from repeated exposure to a sexually transmitted HPV and possibly an additional agent. HPV16 positivity and increased viral load in some earlier studies were predictive of prevalent CIN II/III. More recent studies with more sensitive HPV assays did not corroborate these findings. The role of cigarette smoking is controversial and requires additional study. Accumulating evidence suggests that high risk HPV DNA detection and persistence are predictive of CIN I progressing to CIN II/III. Other possibilities are persistence of a high risk HPV variant, altered cell immunity, and cigarette and oral contraceptive use. Possible biomarkers include aneuploidy, aneusomy of chromosomes 1 and 3, Ras and bcl-2 oncogene over expression, and cytokeratin 13 protein under-expression. PMID- 11890104 TI - Clinical oncology current status in Singapore. PMID- 11890105 TI - Pre-clinical and early-phase clinical studies of curcumin as chemopreventive agent for endemic cancers in Taiwan. PMID- 11890106 TI - Clinical application of two new serological tests of Epstein-Barr virus in nasopharyngeal carcinoma. PMID- 11890107 TI - Liver cancer, the prevention and control in Thailand. PMID- 11890108 TI - Current development of clinical oncology in Japan. PMID- 11890109 TI - Mechanism of action of all-trans retinoic acid and arsenic trioxide in the treatment of acute promyelocytic leukemia. AB - Treatment of APL with ATRA or As2O3 alone or in combination with chemotherapy yields a complete remission as high as 85%-95%, but their mechanisms of action remain unclear. The mechanisms of action underlying ATRA treatment are (1) relocalization of the PML restoration of normal structure of nuclear bodies and degradation of PML-RAR alpha protein via caspase-mediated cleavage and proteosome dependent degradation; (2) conversion of PML-RAR alpha from a transcription repressor (CoR) to a transcription activator (CoA) under therapeutic concentration of ATRA (3) coordinated genes expression induced by ATRA resulting in an elegant and intricate cellular program for the commitment to differentiation. 169 genes were modulated to express, with 100 genes up-regulated and 69 down-regulated. As2O3 exerts its action by dual dose-dependent manner. At higher concentration (1-2 microns/l), it induces apoptosis of the leukemic cells associated with disruption of mitochondrial membrane potential, elevation of caspase-3 and other caspases activity and decline of Bcl-2 expression. At lower concentration (0.1-0.5 micron/l), it triggers differentiation with elevation of CD11b expression accompanied by morphologically partial differentiation. At both concentrations, As2O3 causes degradation of PML-RAR alpha protein implicated probably in its mechanisms of action. PMID- 11890110 TI - Colorectal carcinoma in Nepalese young adults: presentation and outcome. AB - All cases of primary colorectal carcinoma treated at the Department of Surgery, Tribhuvan University Teaching Hospital, Kathmandu during a period of 5 years were retrospectively reviewed in order to promote a greater awareness of the potential for colorectal carcinoma in young adults under 40 years of age. Of the total 91 patients, 26 (28.6%) were younger than 40 years, and this group included more female patients than the older age group. All patients were symptomatic (single or multiple symptoms) for an average period of 7.6 months (range 1 week to 2 years) before seeking medical advice. In the younger group, rectum was the most frequent site of tumors (76.9%) which was significantly higher than in older age group (36.9%). Younger patients were more likely to present with stage III or IV disease (92.3% vs 61.5%) than the older age group (p = 0.001). Moreover, the younger patients had a significantly higher incidence of poorly differentiated and mucinous carcinoma (p = 0.000). Potentially curative resection was performed in only 10 younger patients and most of them had a recurring disease at a median of 11 months. Curative colectomy was more common in > or = 40 age group (29.2% vs 15.4%). The overall 2-year survival rate was significantly lower in younger age group than in the older patients (4% vs 55%, p = 0.0003). PMID- 11890111 TI - Influence of liver dysfunction on dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells. PMID- 11890112 TI - Current state of HIV/AIDS in Taiwan and reappraisal of early diagnostic value of serial and comparative analyses of western blot patterns of HIV infection. AB - Since Taiwan is not a member of the World Health Organization (WHO), the problems of human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) in Taiwan are not well known to the world. In this report we summarize the trend and current status of HIV/AIDS in Taiwan and also analyze Western blot (WB) patterns. The application of serial and comparative WB analyses is important in establishing an early diagnosis of neonatal HIV infection. These analyses will be useful as well in evaluating the clinical status of adults infected with HIV in developing countries. When there are "indeterminate" WBs in adults with recent HIV infection, it is imperative to perform a WB weekly to attain the earliest possible diagnosis and treatment. Usually anti-gag antibodies, most frequently the anti-p24, are followed by anti-env and anti-pol antibodies. When the set point is attained, WB studies may also be repeated if the clinical status changes or if the patients are receiving highly active antiretroviral therapy (HAART). In the advanced stages of HIV infection, usually the anti-gag antibodies will fade first and then disappear, this occurs next in the anti-pol and in the anti-env antibodies. During HAART, viral replication is usually controlled and there will thus be a partial restoration of the immunological function that will induce changes in the antigen-antibody ratio and, in the end, this will result in changes in the WB patterns. In the mid 1980s all the hepatitis B immunoglobulin (HBIG) used in Taiwan was anti-HIV positive. However, follow-up ELISA and WB studies of the HBIG injected newborns proved that HBIG was anti-HIV positive but that there was no replicable HIV in HBIG. Monthly WB tests of newborns and anti HIV positive mothers were used to differentiate HIV infection from the passive placental transference by comparing their WB patterns. Comparative WB analyses was also done between blood donors and recipients, and also between husbands and wives. In this 21st century there are probably more than 36 million people infected with HIV in the world and over 95% of them are in developing countries. In these countries people are much less able to respond to HIV/AIDS crisis. Not only are they unable to obtain such expensive anti-retroviral therapies they are also unable to carry out CD4 and viral load quantitative analyses. However, we believe that each country can afford to carry out WB studies. With experience, the reaction patterns can be interpreted easily and correlated with any clinical changes (Fig. 1). PMID- 11890113 TI - Cancer treatment in Pakistan: challenges & obstacles. AB - INTRODUCTION: There is no population based tumor registry in Pakistan except for Karachi. A Department based tumor registry was established to determine the patterns of cancers in Pakistan especially Punjab. Other objectives were to determine socioeconomic status, disease stage, co-morbid conditions. We also tried to determine if patients had received optimal treatment on diagnosis. MATERIALS & METHODS: 3,274 patients presented to the Department of Oncology between 1995-2000. All patients had histopathologically confirmed diagnosis of cancer. Demographic data included age, sex, socioeconomic status, smoking, other co-morbid and occupation. RESULTS: The Oncology Department drains a catchment area of approximately 400 square kms in Punjab. Breast cancer was the most common cancer in females while leukemias and lymphomas were most common in males. Poor socio-economic status was present in 89% of cases. Illiteracy was present in 76%. Comorbid conditions like hepatitis B & C were present in 37% of patients. Advanced disease was documented in 59% patients. Optimal treatment on initial diagnosis was not provided to 45% patients. DISCUSSION: Advanced stage, poor socio-economic status, illiteracy were common. Associated co-morbid conditions were a major cause in delay in treatment. Optimal treatment was not provided to majority of patients. All these factors are contributing to poor cure rates seen in Pakistan. PMID- 11890114 TI - The Charter of Paris Against Cancer--a major act marking an historical realization and turning point. PMID- 11890116 TI - Novel anti-angiogenetic agents developing in China. PMID- 11890115 TI - Global development of angiogenesis inhibitors for cancer: principles, progress and new paradigms. PMID- 11890117 TI - Recent progress of traditional Chinese medicine and herbal medicine for the treatment and prevention of cancer. PMID- 11890119 TI - Current status of chemotherapy for gastric cancer in Japan. PMID- 11890118 TI - Recent development of anti-cancer drugs for treatment of GI malignancies in Japan. PMID- 11890120 TI - Clinical trials of new anticancer agents in recent years in China. PMID- 11890121 TI - SNU cell lines and their application for cancer research. AB - The cellular and molecular characteristics of SNU human cancer cell lines established in Korean are summarized according to the genetic and epigenetic alterations and functional analysis, etc. We have characterized and reported 89 different cell lines since 1987. These cell lines have been distributed to biomedical researchers through the KCLB (Korean Cell Lines Bank). Some cell lines have several distinguishing characteristics; i) two hepatocellular cell lines (SNU-354 and -368) expressing the MDR1 gene and hepatitis B virus, ii) Gastric carcinoma cell lines with mutations in TGF-beta type II receptor gene, iii) uterine cervical carcinoma cell line (SNU-1000) having intact episomal HPV-16, and iv) ovarian carcinoma cell line (SNU-251) have a nonsense mutation of codon 1815 in exon 23 of the BRCA1 gene. The distributed SNU cell lines have proven to be of value in various scientific research fields. PMID- 11890122 TI - Alternative diets. AB - As pet owners become more conscious of their own diets and the impact it has on their health, they naturally become more interested in what their animal companions are eating and how that might be affecting their pet's health. Many are exploring alternatives to standard commercial pet foods, and some are asking their veterinarians for advice. Small-animal nutrition is an ever-changing field. What veterinarians were taught 10 years ago may no longer be sound advice. This article explores some of the reasoning behind the development of both conventional commercial pet foods and the alternative foods and diets. It questions some of the conventional dogma as well as some of the trendy assumptions in the current marketplace. The intent is to provide the veterinarian with some balanced information on which to base nutritional advice to clients, and to begin forming new opinions or at least asking new questions. Guidelines on formulating a homemade diet for dogs are included. PMID- 11890123 TI - Complementary and alternative veterinary medicine and gastrointestinal disease. AB - Gastrointestinal, hepatic, and pancreatic diseases provide a significant challenge to the veterinary practitioner. Specific causes and effective therapies can be elusive and sometimes frustrate both the animal caretaker and the veterinarian. The therapeutic options of a conventional veterinary practice are frequently limited and may come down to a decision of which is worse: the disease or the side effects of the treatment. This article provides information for the veterinary practitioner to consider for expanding his/her options. Acupuncture, herbal remedies, and homeopathy are not newly discovered modalities. They are old practices that are getting a fresh look from Western medicine as we seek new ways to better serve our patients and clients. The goal of this article is to provide the reader with many ideas and sufficient solid information to consider the use of these options. Specific therapies are suggested for numerous gastrointestinal and liver problems. Many of these may be used in conjunction with conventional therapies to aid in the healing process. PMID- 11890124 TI - Complementary and alternative medicine for neurologic disorders. AB - The use of complementary and alternative veterinary medicine in treating neurologic disorders has increased in popularity in response to advances in human alternative and integrative therapies. Neurolocalization of lesions to the brain, spinal cord, and neuromuscular systems is discussed, as well as the diagnostics and therapeutics used to treat such disorders. Emphasis is placed on integrative and alternative treatments for such neurologic diseases as seizures, cerebrovascular accidents, canine cognitive disorder, meningitis, intervertebral disc disease, fibrocartilagenous embolism, degenerative myelopathy, and myopathies. Thorough physical and neurologic examinations, establishment of a correct diagnosis, and integrative therapeutics are aimed at improving the overall quality of life of the veterinary patient. PMID- 11890125 TI - Complementary and alternative veterinary medicine and urologic conditions. PMID- 11890126 TI - Alternative therapies for pruritic skin disorders. PMID- 11890127 TI - Alternative medicine: musculoskeletal system. PMID- 11890128 TI - Alternative medicine and behavior. PMID- 11890129 TI - Complementary and alternative veterinary medicine: the immune system. PMID- 11890130 TI - Herbal medicine: potential for intoxication and interactions with conventional drugs. AB - The use of herbal remedies for the prevention and treatment of a variety of illnesses in small animals has increased tremendously in recent years. Whereas most herbal remedies, when used as directed and under the supervision of knowledgeable individuals, are safe, the potential for adverse effects or intoxications certainly exists. Due to inherent toxicity, some herbal remedies should not be used under any circumstance. In addition, because nearly all herbal remedies contain multiple, biologically active constituents, interaction with conventional drugs is a concern. It is incumbent upon clinicians to be aware of those herbs that can cause intoxication, and to be cognizant of potential herb drug interactions. There are a number of evidence-based resources available to assist clinicians in the safe use of herbal remedies. PMID- 11890131 TI - A unique wound dressing and a case of misclassification. PMID- 11890132 TI - The high price of cutting costs. PMID- 11890133 TI - Legalism and ethics in healthcare. AB - Legalism is the translation of moral problems into legal problems--the repression of moral debate for fear it will be translated into a legal mandate. Some assert that legalism interferes with the serious debate of significant moral issues, while others believe that it elevates moral judgments to moral standards. The purpose of this article is to describe legalism and its impact on ethical decision-making, particularly in the wound care setting. PMID- 11890134 TI - Pelvic organ prolapse: a review. AB - Pelvic organ prolapse is a common medical problem in parous women. This condition usually refers to a combination of deficiencies of the pelvic organs as they relate to support mechanisms of the vaginal wall. Symptoms vary--an accurate diagnosis requires a careful and complete physical examination with attention directed toward the pelvis and perineum. Although many patients will not require surgical treatment for pelvic organ prolapse, a comprehensive approach to repair in which all of the anatomic defects affecting support are addressed is necessary for successful treatment. Patients presenting with pelvic organ prolapse often provide some of the most complex, challenging, and rewarding cases in reconstructive pelvic surgery. This article addresses the definitions and classifications, prevalence and risk factors, and anatomy and pathophysiology relevant to pelvic organ prolapse. Discussion also includes diagnosis and approaches to management (surgical and nonsurgical) of anterior vaginal wall prolapse, cystourethrocele, apical vaginal prolapse, uterine prolapse and enterocele, posterior vaginal wall prolapse, rectocele, and pelvic floor relaxation and perineal laxity, with indications for and approaches to surgery, along with possible complications. PMID- 11890135 TI - Pessary placement and management. AB - Female pelvic organ prolapse is a common and aggravating condition that few women openly discuss. Fortunately, nonsurgical and surgical treatment options exist for this condition. Nonsurgical treatments include pessaries, which are used to reduce prolapse. The type of pessary depends on the direction and extent or grade of the protruding organ. The use of vaginal pessaries for urinary stress continence is a relatively new treatment option. Proper fit that is determined by trial is essential for effectiveness. Nurses at all levels now need to be familiar with the use of the vaginal pessary for pelvic organ prolapse and urinary stress incontinence. Nurses' roles regarding pessary use need to be clearly defined. Continence nurses should be familiar with the indications for pessary use, the types available for incontinence, and patient education regarding pessary care. Home health and long-term care nurses are frequently asked to periodically remove and insert pessaries. Skills and tips for making removal and insertion as easy as possible need to be disseminated. Advance practice nurses need skills for assessment of prolapse, fitting, and current outpatient pessary care routines. Increasing nurses' understanding of and comfort with pessary use might make a significant difference in the treatment of pelvic organ prolapse and urinary stress incontinence. PMID- 11890136 TI - Pelvic disorders in women: chronic pelvic pain and vulvodynia. AB - Chronic pelvic pain and vulvodynia are frustrating pelvic disorders seen in young adult women. In the medical literature, these two conditions are linked together under the category of "chronic pelvic pain syndromes." Underlying pathophysiology is not well understood, and relatively scant research is available on successful treatment options. Patients often seek the help of specialists who provide nonsurgical treatments for incontinence and related pelvic disorders. This article provides an overview of the clinical presentation of both chronic pelvic pain and vulvodynia. Specific evaluation techniques, including abdominal, pelvic, bimanual rectal-vaginal, and neurologic examinations, are described. Several practical treatments, such as dietary interventions, vitamin supplementation, muscle relaxation training, biofeedback therapy, and electrical stimulation are discussed as options in a private practice setting. PMID- 11890137 TI - Assessing and treating patients with complex fecal incontinence. AB - Fecal incontinence is the involuntary loss of gas, liquid, and/or solid stool. It affects 2.2% of the general population. Because fecal incontinence can be socially and psychologically devastating, and is not easily discussed, this figure is probably understated. Patients presenting with fecal incontinence need to be properly assessed, including physiological testing of the pelvic floor muscles and nerves. Identifying any abnormal anatomy or physiology in the pelvic floor helps the clinician develop a care plan that best suits the patient's etiology. Knowledge of the physiology of the pelvic floor musculature and its effects on continence is improving. Treatment options also have broadened. This article describes the current techniques of assessment and treatment, including the "gold standard," and newer investigational procedures offered to patients with complex fecal incontinence. PMID- 11890138 TI - Abstracts from the 5th International Brain and Cardiac Surgery Conference at the Royal Society of Medicine. London, United Kingdom, 7-9 September 2000. PMID- 11890139 TI - Proceedings of the Titus H. J. Huisman Memorial Symposium. Augusta, Georgia, USA. June 9, 2000. PMID- 11890140 TI - Proceedings of the 20th Symposium on Capillary Electrophoresis. Hyogo, Japan. November 29-December 1, 2000. PMID- 11890141 TI - Proceedings of the Special IBMTR/ABMTR Workshop on the Role of Hematopoietic Stem Cell Transplantation for Rheumatoid Arthritis. Anaheim, California, USA. March 26, 2000. PMID- 11890144 TI - Karlin v. Foust. PMID- 11890145 TI - 2001 year in review. ACHE (American College of Healthcare Executives): our focus is on you. PMID- 11890146 TI - The role of the hospitalist. PMID- 11890147 TI - An evaluation of the minimum-jerk and minimum torque-change principles at the path, trajectory, and movement-cost levels. AB - In the present study we evaluated the minimum jerk and the minimum torque-change model at the path, trajectory, and movement-cost levels. To date, most evaluations of these models have mainly been restricted to path comparisons. Assessments of the time courses of realized jerk and torque changes are surprisingly lacking. Moreover, the extent to which the presumed optimized parameters change as a function of the duration and other temporal features of aiming movements has never been investigated, most probably because the models presuppose movement time. In order to fill this gap, we analyzed a subset of the data of an earlier experiment in which 12 participants performed leftward and rightward planar pointing movements. Hand displacements and joint excursions were recorded with a 3D motion-tracking system and subsequently evaluated by means of model-based analyses. The results show that despite a good agreement between observed paths and predicted paths, especially by the minimum torque-change model, the time courses of jerk and torque changes of observed and modeled movements differed considerably. These differences could mainly be attributed to asymmetrical properties of the time functions of slow movements. Variations of movement costs as a function of movement time and skewness of tangential velocity profiles show that, especially at high movement speed, costs increase exponentially with departures of symmetry. It is concluded that trajectory formation models have limited explanatory power in situations that require demanding information processing during the homing-in phase of goal-directed movements. However, for slow movements, deviations from the optimal timing profiles require little extra costs in terms of jerk or torque change. PMID- 11890148 TI - Separate sources of spatial information for distance and location in rapid aiming movements. AB - The purpose of this research was to examine the role of distance and location information in the production of rapid aiming movements. Participants performed an aiming task consisting of horizontal left-handed elbow flexion movements that translated to movements of a cursor on an oscilloscope screen. The location of the home position and the target on the oscilloscope screen were fixed but the initial angle of the elbow was varied randomly. Participants were informed that the required distance was always constant. Initial impulse and error correction phases were analyzed to examine whether separate spatial codes for distance and position were used in the control of these two movement phases. The results indicated that initial impulse endpoints and the final positions of the limb overshot the target from the leftmost starting positions, while they undershot the target from the rightmost starting positions. Also, varying the initial angle of the elbow had a greater influence on the final position of the limb than initial impulse endpoints. PMID- 11890149 TI - Peter L. Steponkus memorial issue. PMID- 11890150 TI - Proceedings of the British Electrophoresis Society Meeting. York, United Kingdom, 4-6 April 2001. PMID- 11890151 TI - Bibliography. Current world literature. Gynecologic oncology and pathology. PMID- 11890152 TI - [Subjective element in psychiatry]. PMID- 11890153 TI - Re: Belohlakova et al. Fenfluramine-induced pulmonary vasoconstriction: role of serotonin receptors and potassium channels. J Appl Physiol 91:755-761, 2001. PMID- 11890154 TI - C1-C2 transarticular screw fixation for atlantoaxial instability: a 6-year experience. PMID- 11890155 TI - Caspase-9 transduction overrides the resistance mechanism against p53-mediated apoptosis in U87MG glioma cells. PMID- 11890156 TI - Patent forman ovale as a possible risk factor for cryptogenic brain abscess: report of two cases. PMID- 11890157 TI - Endoscopic endonasal approaches to the cavernous sinus: surgical approaches. PMID- 11890158 TI - Bibliography. Current world literature. Lipid metabolism and therapy. PMID- 11890159 TI - Bibliography. Current world literature. Nutrition in the intensive care unit. PMID- 11890160 TI - Effectiveness of anorectal biofeedback therapy for faecal incontinence: medium term results. PMID- 11890161 TI - Manually operated. PMID- 11890162 TI - Animal models of insulin resistance and cardiovascular disease: some therapeutic approaches using JCR:LA-cp rat. PMID- 11890163 TI - Minor long-term changes in weight have beneficial effects on insulin sensitivity and beta-cell function in obese subjects. AB - AIM: To evaluate the long-term effect of changes in body composition induced by weight loss on insulin sensitivity (SI), non-insulin mediated glucose disposal, glucose effectiveness (SG)and beta-cell function. DESIGN: Glucose metabolism was evaluated before and after participation in a two-year weight loss trial of Orlistat vs. placebo, combined with an energy and fat restricted diet. SUBJECTS: Twelve obese patients (11 women, 1 man), age 45.8 +/- 10.5 years, body weight (BW) 99.7 +/- 13.3 kg, BMI 35.3 +/- 2.8 kg/m(2). MEASUREMENTS: At inclusion and 2 years later an oral glucose tolerance test (OGTT) and a frequently sampled intravenous glucose tolerance test (FSIGT) were performed. Body composition was estimated by a dual-energy X-ray absorptiometry (DXA) whole body scanning. RESULTS: The patients obtained varying changes in BW ranging from a weight loss of 17.8 kg to a weight gain of 6.0 kg. Corresponding changes in fat mass (FM) varied from a 40% reduction to a19% increase. A significant decrease in both fasting (p = 0.038) and 2 h (p = 0.047) blood glucose at OGTT was found. The improvement in insulin sensitivity (SI) estimated by means of Bergmans Minimal Model, was significantly and linearly correlated to change in total FM (r = - 0.83,p = 0.0026). A multiple regression analysis showed that changes in truncal FM was the strongest predictor of change in S(I) explaining 67% of the variation. First phase insulin response (AIRg)remained unchanged whereas insulin disposition index increased significantly (p = 0.044). At inclusion five patients had impaired glucose tolerance of which four, who lost weight, were normalized at the retest 2 years later. CONCLUSION: In obese subjects long-term minimal or moderate changes in weight were found to be linearly associated with changes in insulin sensitivity. In obese subjects with impaired glucose tolerance even a minor weight loss was able to normalize glucose tolerance. PMID- 11890164 TI - Annual survey documents continued increases in use of ecstasy among U.S. teenagers. PMID- 11890165 TI - Measuring portal venous perfusion with contrast-enhanced CT: comparison of direct and indirect methods. AB - RATIONALE AND OBJECTIVES: Two algorithms can be used to measure portal venous perfusion (PVP) with contrast material-enhanced single-level liver computed tomography. The "direct" and "indirect" algorithms use data from the portal vein and aorta, respectively. This study compared PVP values obtained with direct and with indirect algorithms in a series of patients. MATERIALS AND METHODS: Both techniques were applied in 27 patients with cirrhosis (14 men and 13 women; mean age, 56.1 years +/- 9.4) and 18 control patients (seven men and 11 women; 52.8 years +/- 12.3). A single section through the liver was scanned after intravenous injection of ioversol (40-mL bolus; 320 mg of iodine per milliliter). RESULTS: Both techniques showed reduced PVP in patients with cirrhosis (0.63 for direct and 0.17 for indirect method) compared with control patients (1.06 and 0.26, respectively), but only the direct method agreed with physiologic expectations based on animal and human studies. In separating cirrhotic and control patients, the area under the receiver operating characteristic curve was significantly greater for the direct method (0.91 vs 0.78; P = .03). CONCLUSION: Both direct and indirect methods are feasible and distinguish well between cirrhotic and control patients, but the direct method is more physiologic and is preferable if portal venous data are available. PMID- 11890166 TI - Selecting a probability distribution for extreme rainfall series in Malaysia. AB - This paper discusses the comparative assessment of eight candidate distributions in providing accurate and reliable maximum rainfall estimates for Malaysia. The models considered were the Gamma, Generalised Normal, Generalised Pareto, Generalised Extreme Value, Gumbel, Log Pearson Type III, Pearson Type III and Wakeby. Annual maximum rainfall series for one-hour resolution from a network of seventeen automatic gauging stations located throughout Peninsular Malaysia were selected for this study. The length of rainfall records varies from twenty-three to twenty-eight years. Model parameters were estimated using the L-moment method. The quantitative assessment of the descriptive ability of each model was based on the Probability Plot Correlation Coefficient test combined with root mean squared error, relative root mean squared error and maximum absolute deviation. Bootstrap resampling was employed to investigate the extrapolative ability of each distribution. On the basis of these comparisons, it can be concluded that the GEV distribution is the most appropriate distribution for describing the annual maximum rainfall series in Malaysia. PMID- 11890167 TI - Regional frequency analysis of extreme rainfalls. AB - This study proposes two alternative methods for estimating the distribution of extreme rainfalls for sites where rainfall data are available (gaged sites) and for locations without data (ungaged sites). The first method deals with the estimation of short-duration rainfall extremes from available rainfall data for longer durations using the "scale-invariance" concept to account for the relationship between statistical properties of extreme rainfall processes for different time scales. The second method is concerned with the estimation of extreme rainfalls for ungaged sites. This method relies on a new definition of homogeneous sites. Results of the numerical application using data from a network of 10 recording rain gauges in Quebec (Canada) indicate that the proposed methods are able to provide extreme rainfall estimates that are comparable with those based on observed at-site rainfall data. PMID- 11890168 TI - Sensitive liquid chromatography-tandem mass spectrometry method for the determination of clarithromycin in human plasma. AB - A sensitive method for the determination of clarithromycin in plasma is described, using high-performance liquid chromatographic separation with tandem mass spectrometric detection. Samples were prepared using liquid-liquid extraction and separated on a Supelco Discovery C18 column with a mobile phase consisting of acetonitrile, methanol and acetic acid. Detection was performed by a PE SCIEX API 2000 mass spectrometer in the multiple reaction monitoring (MRM) mode (LC-MS-MS) using TurbolonSpray ionization and monitoring the transition of the protonated molecular ion for clarithromycin at m/z 748.5 (M+1) to the predominant product ion of m/z 158.2. The mean recovery of clarithromycin was 87.3%, with a lower limit of quantification of 2.95 ng/ml when using 0.3-ml plasma. This high-throughput method was used to quantify 230 samples per day, and is sufficiently sensitive to be employed in pharmacokinetic studies. PMID- 11890169 TI - Determinants of adaptive behavior among older persons: self-efficacy, importance, and personal dispositions as directive mechanisms. AB - Successful aging calls for effective adaptation, which in turn implies flexible use of coping strategies to optimize personal functioning and well-being. The present paper studied adaptive choice behavior of older, independently living persons faced with complications in their houses. The goal was to gain insight into the concrete coping process and its outcome-in terms of the choice of assimilative vs. accommodative strategies-and in the role of three determinants on this process. The determinants were perceived self-efficacy, importance of the problem, and personal dispositions (flexibility and tenacity). A sample of 199 independently-living older persons participated in an experiment that was based on a scenario and questionnaire method, with problems stemming from the domain of independent living. Results mainly underlie the crucial role of perceived self efficacy and are discussed in view of the concept of successful aging. PMID- 11890170 TI - Predictors of grandmother participation in a multigenerational study. AB - Results of a growing body of literature regarding multigenerational relationships indicate that grandparents directly and indirectly influence their grandchildren's development. However, multiple generations have been difficult to assemble for research purposes, with refusal rates ranging from 40 percent to over 80 percent. Despite difficulties in obtaining information from more than one generation of family members, data examining potential biases in the selection of participants have rarely been reported. The present study explored predictors of grandmother participation in a multigenerational study with 84 mothers, 84 infants, and 52 maternal grandmothers. Results indicated that grandmothers were less likely to participate in the study when grandchildren experienced preterm birth and when mothers reported less family of origin support. However, grandmothers who participated in the study reported high levels of intergenerational involvement and contact with daughters and grandchildren. PMID- 11890171 TI - Looking inward: introspectiveness, physical disability, and depression across the life course. AB - This study investigates the interrelationships among age, physical disability, introspectiveness, and depression. Using data from a community sample of disabled and non-disabled adults (N = 1,567), this study tests: 1) if there are age variations in introspectiveness; 2) if age variations in introspectiveness differ by physical disability status; 3) if introspectiveness mediates the association between age and depression; 4) if introspectiveness and disability status have synergistic effects on depression; and if so, 5) if subjective health differences between disabled and nondisabled account for the joint impact of introspectiveness and disability status on depression. Results show that older people report less introspectiveness than younger people do--which explains part of the negative association between age and depression. Additionally, the negative association between age and introspectiveness is significantly stronger among nondisabled respondents. Adjustment for less introspectiveness among older adults accounts for about 24 percent of the negative association between age and depression. Disabled respondents experience a more positive relationship between introspectiveness and depression; however, disabled respondents' poorer global health explains most of that pattern. PMID- 11890172 TI - The cumulative effects of life event, personal and social resources on subjective well-being of elderly widowers. AB - A multidimensional Life Stress Model was used to test the independent contributions of background characteristics, personal resources, life event, and environmental influences on 200 widowers' levels of well-being, measured by the Affect Balance Scale. Stepwise regression analyses revealed that environmental resources were unrelated to negative affect which is influenced more by the life event and personal resource variables. The environmental resource variables, particularly interactions with friends and neighbors, mostly influenced positive affect. The explanatory model for well-being included multiple variables and explained 33 percent of the variance. Although background characteristics had the greatest impact, absence of hospitalization, higher mastery, higher self-esteem, contacts with friends, and interaction with neighbors enhanced well-being. The results support previous speculations on the importance of positive exchanges for positive affect. African-American widowers showed higher levels of well-being than Caucasian widowers did. The results advance knowledge about differences among elderly men. PMID- 11890173 TI - Social support exchange among elderly Chinese people and their family members in Hong Kong: a longitudinal study. AB - This study examines the pattern of social support exchange between the elderly and their family members, focusing on financial aid and household care (both instrumental and emotional). The second objective of this study is to identify resource capacity factors (including education, physical health, and size of social network) that generate differences in the exchange of financial aid and household care between elderly people and their family members. The respondents were 213 people who had family members living in Hong Kong aged 70-years-old or older from a longitudinal study of a representative community sample of the elderly population in Hong Kong. Using multiple regression models, we found that the elderly subjects who received more household care from their family members were likely to provide more household care to their family members three years later, even after controlling for the impact of resource capacities; and the elderly subjects who provided more household care to their family members were more likely to receive more household care from their family members three years later. In assessing the impact of the resource variables on support exchange, functional disability and the number of close relatives were significantly associated with the amount of household care the old people provided, whereas the number of close family members were significantly associated with the amount of household care the old people received. Policy implications of the findings in this study were discussed. PMID- 11890174 TI - Biochemical changes induced by hypomagnesaemia in lactating cows and ewes. AB - Severe hypomagnesaemia was induced in lactating cows and lactating sheep by feeding them magnesium-deficient diets for 17 and 14 days, respectively. Hypomagnesaemia in cows was associated with abnormally high rates of change in the numbers of leucocytes, neutrophils, monocytes and platelets. There were increases in the concentration of iron in the liver of the hypomagnesaemic ewes and in the heart of the hypomagnesaemic cows, which were not associated with a haemolytic process. The percentage of some of the peroxidisable fatty acids was lower in the heart tissue of hypomagnesaemic cows, but the reduction was not associated with significant lipid peroxidation. PMID- 11890175 TI - Supervisor tolerance-responsiveness to substance abuse and workplace prevention training: use of a cognitive mapping tool. AB - Supervisor tolerance-responsiveness, referring to the attitudes and behaviors associated with either ignoring or taking proactive steps with troubled employees, was investigated in two studies. The studies were conducted to help examine, understand and improve supervisor responsiveness to employee substance abuse. Study 1 examined supervisor response to and tolerance of coworker substance use and ways of interfacing with the Employee Assistance Program (EAP) in two workplaces (n = 244 and 107). These surveys suggested that engaging supervisors in a dialogue about tolerance might improve their willingness to use the EAP. Study 2 was a randomized control field experiment that assessed a team oriented training. This training adopted a cognitive mapping technique to help improve supervisor responsiveness. Supervisors receiving this training (n = 29) were more likely to improve on several dimensions of responsiveness (e.g. likely to contact the EAP) than were supervisors who received a more didactic, informational training (n = 23) or a no-training control group (n = 17). Trained supervisors also showed increases in their own help-seeking behavior. Procedures and maps from the mapping activity (two-stage conversational mapping) are described. Overall, results indicate that while supervisor tolerance of coworker substance use inhibits EAP utilization, it may be possible to address this tolerance using team-oriented prevention training in the work-site. PMID- 11890176 TI - The Sexual Health Model: application of a sexological approach to HIV prevention. AB - This article outlines the Sexual Health Model and its application to long-term HIV prevention through comprehensive, culturally specific, sexuality education. Derived from a sexological approach to education, the model defines 10 key components posited to be essential aspects of healthy human sexuality: talking about sex, culture and sexual identity, sexual anatomy and functioning, sexual health care and safer sex, challenges to sexual health, body image, masturbation and fantasy, positive sexuality, intimacy and relationships, and spirituality. A brief review of literature supporting a need for a more explicit focus on sexuality and relationships in HIV prevention is presented to demonstrate the relevance of the Sexual Health Model. The model in anchored in a holistic definition of sexual health. This definition is followed by a description of the Sexual Health Model's developmental origins in sexuality education, the importance of culturally relevant information, and the authors' qualitative and quantitative research. The model's 10 key components are discussed in more depth, and the theoretical and practical applications of this approach to HIV prevention are discussed. The article concludes with some cautions and suggestions for research. It is recommended that HIV prevention agencies contemplating use of the model should design their sexual health intervention to fit the unique needs of their target population. Evaluation of the effectiveness of interventions based on the model has begun, but further research is needed to confirm its viability. PMID- 11890177 TI - The detection and identification of quaternary nitrogen muscle relaxants in biological fluids and tissues by ion-trap LC-ESI-MS. AB - Quaternary nitrogen muscle relaxants pancuronium, rocuronium, vecuronium, gallamine, suxamethonium, mivacurium, and atracurium and its metabolites were extracted from whole blood and other biological fluids and tissues by using a solid-phase extraction procedure. The extracts were examined by using high performance liquid chromatography-electrospray ionization mass spectrometry (LC ESI-MS). The drugs were separated on a ODS column in a gradient of ammonium acetate buffer (pH 5.0) and acetonitrile. Full-scan mass spectra of the compounds showed molecular ions, and MS-MS spectra showed fragments typical of the particular compounds. LC-ESI-MS allowed an unequivocal differentiation of all muscle relaxants involved. The method was applied in a case of rocuronium and suxamethonium administration in a Caesarian section and in a case of intoxication by pancuronium injection. In both cases, the administered drugs could be detected and identified in the supplied samples. PMID- 11890178 TI - Quantitative determination of n-propane, iso-butane, and n-butane by headspace GC MS in intoxications by inhalation of lighter fluid. AB - This report describes a fully elaborated and validated method for quantitation of the hydrocarbons n-propane, iso-butane, and n-butane in blood samples. The newly developed analytical procedure is suitable for both emergency cases and forensic medicine investigations. Its practical applicability is illustrated with a forensic blood sample after acute inhalative intoxication with lighter fluid; case history and toxicological findings are included. Identification and quantitation of the analytes were performed using static headspace extraction combined with gas chromatography-mass spectrometry. In order to reconcile the large gas volumes injected (0.5 mL) with the narrowbore capillary column and thus achieve preconcentration, cold trapping on a Tenax sorbent followed by flash desorption was applied. Adequate retention and separation were achieved isothermally at 35 degrees C on a thick-film capillary column. Sample preparation was kept to a strict minimum and involved simply adding 2.5 microL of a liquid solution of 1,1,2-trichlorotrifluoroethane in t-butyl-methylether as an internal standard to aliquots of blood in a capped vial. Standards were created by volumetric dilution departing from a gravimetrically prepared calibration gas mixture composed of 0.3% of n-propane, 0.7% of iso-butane, and 0.8% of n-butane in nitrogen. In the forensic blood sample, the following concentrations were measured: 90.0 microg/L for n-propane, 246 microg/L for iso-butane, and 846 microg/L for n-butane. PMID- 11890179 TI - A global experiment under way. PMID- 11890180 TI - Modeling macroscopic patterns in ecology. PMID- 11890181 TI - Section 8 project-based rental assistance: the potential loss of affordable federally subsidized housing stock. PMID- 11890183 TI - Effects of age on physiological and psychological functioning among COPD patients in an exercise program. AB - Older adults with chronic illness may be at greater risk of psychological distress than healthy older adults. This study examined the effect of age on physiological and psychological functioning of exercising older adults with chronic obstructive pulmonary disease (COPD). Sixty-four older adults (mean age = 67.4 plus or minus 7.0; 35 male, 29 female) with COPD were included in the study. All subjects participated in a 30-day rehabilitation program consisting of exercise (walking, stationary bicycle, pool exercises), education, and stress management. Subjects underwent assessments of physiological functioning (e.g., bicycle ergometry,12-minute walk), cognitive status (e.g., memory, psychomotor speed, concentration), and psychological well-being (e.g., anxiety, depression, psychiatric symptoms) before and after the exercise program. Results were analyzed by repeated measures multivariate analysis of variance, with age (median split = 67.5 years) as a between-subject factor. Although the exercise program appeared to have a greater effect on physiological functioning and on concentration and short-term memory of younger-old subjects, both groups of subjects achieved gains in physiological functioning, and psychological well being. PMID- 11890182 TI - Serum antioxidant status of civil aircrew. AB - Objective. To study the serum antioxidant status in civil aircrew members who had more than 4000 h of cumulative flight hours, therefore exposed to a higher dose of cosmic radiation comparing to the dose received by ground residents. Method. Differences in the serum levels of total antioxidant capacity (TAOC), superoxide dismutase (SOD) and malondialdehyde (MDA) were investigated in 230 crew members, 37 local ground residents in Xinjiang, and 37 ground residents in Tianjin. Result. Significantly higher levels of all the three serum indexes were found in civil aircrew members. Serum levels of TAOC and SOD were higher in Xinjiang than in Tianjin ground residents, although no difference was found for MDA. Positive correlations were observed among the three serum indexes in this study. Conclusion. The elevated serum level of MDA in civil aircrew members deserves a proper attention by health care policy makers. PMID- 11890184 TI - Older Floridians' attitudes toward and use of dental care. AB - Older Floridians (mean age 78 years) were interviewed regarding their use of dental care, attitudes toward dental care, and other characteristics hypothesized as being explanatory of dental care use. Fifty-two percent of respondents reported having been to a dentist within the past year, and 31% had not been within the previous 5 years. Five constructs measured attitudes toward dental care and dental health: (a) the importance placed on regular dental care and oral hygiene, (b) the importance of avoiding tobacco to prevent oral cancer, (c) the value of dental care, (d) negative aspects of dental care, and (e) satisfaction with the last dental visit. In a multivariate model, the value of dental care and importance of regular care and oral hygiene wer significantly correlated with dental care use. These findings are consistent with the conclusion that attitudes contribute to understanding dental care use in later life, a contribution that is independent of the direct effects of socioeconomic status and dentate status. PMID- 11890185 TI - Effects of reboxetine on Hamilton Depression Rating Scale factors from randomized, placebo-controlled trials in major depression. AB - Reboxetine is the first selective norepinephrine reuptake inhibitor (NRI) approved for the treatment of major depressive disorder (MDD). Although reboxetine has demonstrated efficacy for the treatment of depression, its effects on specific depressive symptoms have not been reported. We evaluated the effects of reboxetine on four Hamilton Depression Rating Scale (HAM-D) factors: psychomotor retardation, anxiety, cognitive disturbance and insomnia. Data were obtained from four short-term (4-8-week), randomized, placebo-controlled trials of reboxetine for the treatment of MDD. For each study, mean changes in HAM-D symptom factor scores from randomization to the study endpoint were compared between reboxetine and placebo. In addition, data from all four studies were pooled to determine the proportions of patients who either improved or worsened with treatment were compared between placebo (n = 353) and reboxetine (n = 350) treatment groups. Compared to placebo, reboxetine significantly improved psychomotor retardation in all four trials. Cognitive disturbance and anxiety were improved in three of four trials, and insomnia was improved in one trial with a positive trend in the second trial. Reboxetine, a selective NRI, improves symptoms of psychomotor retardation, anxiety and cognitive disturbance during treatment of MDD. PMID- 11890186 TI - Differential effects of milnacipran and fluvoxamine, especially in patients with severe depression and agitated depression: a case-control study. AB - We attempted to compare the antidepressant efficacy of milnacipran and fluvoxamine in 202 outpatients with major depression, using the 17-item Hamilton Depression Rating Scale (HDRS). Special attention was paid to the difference of responsiveness as a function of the severity of depression and individual HDRS factors. As a result, while no significant difference between the treatment groups was found overall, a positive response (50% or more decrease in total score from the baseline) was recorded significantly more often with milnacipran than fluvoxamine recipients whose baseline HDRS total score was greater than 19 points. Furthermore, there was a significant difference of response for the 'agitation' and 'insomnia' factors in favour of milnacipran. In both treatment groups, the incidence of adverse events, characteristic of tricyclic antidepressants such as dry mouth, constipation, somnolence and postural hypotension, was low. While complaints concerning the upper intestinal tract, such as epigastric distress, were predominant in the fluvoxamine group, urological complications and palpitations were reported only in the milnacipran group. In conclusion, we suggest that milnacipran is preferred to selective serotonin reuptake inhibitors for the treatment of depressed patients with agitation as well as severely depressed patients. PMID- 11890187 TI - Mianserin or placebo as adjuncts to typical antipsychotics in resistant schizophrenia. AB - The beneficial effect of atypical antipsychotic drugs (APDs) in treatment resistant schizophrenia patients has been attributed, mostly, to their relatively high serotonergic (5-HT)2 to dopaminergic (D)2 receptor blockade ratio. We hypothesized that a combination of typical APDs (D2 antagonists) and mianserin, a potent 5-HT2 antagonist, might also exert superior efficacy in this population. Eighteen inpatients with treatment-resistant schizophrenia who had an acute psychotic exacerbation of the disorder received, in a double-blind design, 30 mg/day mianserin (n = 9) or placebo (n = 9) in conjunction with typical neuroleptics [haloperidol (n = 9) or perphenazine (n = 9)]. Clinical status was evaluated before, during, and at the end of 6 weeks of combined treatment with the Brief Psychiatric Rating Scale (BPRS), Scale for the Assessment of Positive Symptoms (SAPS), Scale for the Assessment of Negative Symptoms and Hamilton Rating Scale for Depression. The typical APD/mianserin group exhibited significantly greater improvement in total BPRS scores (17.6% versus 5.5%; P= 0.03) and a trend towards greater improvement in SAPS scores (35.3% versus 13.0%; P = 0.07). Our study indicates that patients with chronic treatment-resistant schizophrenia who have an acute psychotic exacerbation ('acute-on-chronic') may benefit from the addition of a potent 5-HT2 blocker, such as mianserin, to typical antipsychotics. Our findings may further emphasize the contribution of enhanced 5-HT2 blockade to the 'atypicality' of the atypical APDs and to their greater efficacy in alleviating symptoms of chronic treatment-resistant schizophrenia. PMID- 11890188 TI - Time to study discontinuation, relapse, and compliance with atypical or conventional antipsychotics in schizophrenia and related disorders. AB - To more clearly clarify the efficacy of the atypical antipsychotics compared to conventional antipsychotics, we add data on the outcome of patients diagnosed with schizophrenia from two large, international clinical trials comparing olanzapine with haloperidol (n = 1996) and olanzapine with risperidone (n = 339). Both studies comprised double-blinded, placebo controlled, random assignment trials. Health outcomes reported include: (i) time to discontinuation in the trial; (ii) clinical relapse; and (iii) time to drug non-compliance. When outcome was measured as time to discontinuation due to adverse events or lack of efficacy, olanzapine showed superiority over haloperidol and no difference compared to risperidone. Of those patients who had an initial response, there was no significant difference between olanzapine and haloperidol when outcome was measured using either: (i) 52-week relapse rates or (ii) time to first non compliance. Using the measures of study discontinuation, relapse and non compliance, in one trial the atypical antipsychotic olanzapine was superior to haloperidol, while in a second trial there were no differences between olanzapine and risperidone. PMID- 11890189 TI - Rivastigmine in outpatient services: experience of 114 neurologists in Austria. AB - We performed an open-label study in Alzheimer patients on the use of rivastigmine in clinical routine. We evaluated the mode of dosing, safety and response to treatment over 24 weeks in 529 Alzheimer patients recruited by 114 neurologists. At the start of the study, and after 12 and 24 weeks, the participants were tested with the Mini-Mental State Examination and the Global Deterioration Scale. Titration of rivastigmine was slower than expected: After 2 weeks, only 41.8% of subjects received an effective daily dose of at least 6 mg. A total of 33.8% of patients experienced 305 mostly transient adverse events of mild or moderate intensity. Gastrointestinal symptoms occurred in 30.4% of subjects and symptoms of the nervous system in 16.4%. Similar to controlled trials, the cognitive performance of patients improved, with more than 60% of subjects showing improvement from baseline on the Mini-Mental State Examination by week 24. More than 40% of patients showed improvement on the Global Deterioration Scale. This study suggests that, in daily clinical practice, titration of rivastigmine deviates from the prescribing information. Under routine conditions, the safety profile of the drug is favourable, as is the response to treatment. PMID- 11890190 TI - Olanzapine augmentation in the treatment of melancholia: the trajectory of improvement in rapid responders. AB - It has been suggested that the atypical antipsychotic drugs may induce a rapid improvement in individuals with melancholic depression. This might reflect a specific or non-specific effect on mood and other parameters. To examine this issue, ten consecutive patients, for whom olanzapine augmentation was judged a clinically appropriate strategy, were asked to complete daily ratings of depression severity and component features for 1 week. Of the six rapid responders, there was a 52% group improvement in the first day and 89% improvement at 1 week. Item analyses suggested the most rapid impact was evident for insomnia, compared to a slower and linear improvement in depressed mood. Such findings may assist in an understanding of the mechanisms underlying augmentation with atypical neuroleptic drugs. PMID- 11890191 TI - High-dose olanzapine in Huntington's disease. AB - The few reports available on olanzapine in Huntington's disease (HD) are insufficiently documented and/or insufficiently dosed. We describe a 30-year-old woman with genetically confirmed HD who presented with severe chorea. She was not able to eat or dress without help and did not respond to haloperidol; the motor scale of the Unified HD Rating Scale (UHDRS-I) revealed 65 of a possible 124 points. After admission, we treated the patient with a high dose of olanzapine (30 mg daily). The chorea almost ceased in the next 2 days, she was able to eat and walk without assistance (UHDRS-I of 21 points), and fine motor tasks improved, as well as gait and eye movements. This effect lasted for 5 months. We conclude that high-dose olanzapine appears to be useful in grave choreatic attacks. PMID- 11890192 TI - The pattern that connects. AB - Debate over whether nursing is an art or a science culminates in the need for integration of the two as a guide to practice. The historical development of nursing knowledge reveals a spectrum of evolution from physical care to interpersonal relationships to an integrative approach and, most recently, to a unitary perspective. The author proposes pattern as the integrating factor that eliminates the dichotomies of traditional art and science and transforms nursing knowledge to a higher dimension that includes and transcends the knowledge that has gone before. Nursing praxis is presented as integrated theory-research practice that is consistent with a unitary perspective. PMID- 11890193 TI - Coming to know ourselves as community through a nursing partnership with adolescents convicted of murder. AB - This research applies Newman's hermeneutic-dialectic method of pattern recognition to the lives of 12 adolescent males convicted of murder who were invited to be co-investigators in the process of understanding patterns of meaningful relationships and experiences in their lives. Comparison of the 12 life patterns revealed a strikingly similar experience of interactions with the community and yielded insight into community pattern. The process of pattern recognition was found to be transformative. This article proposes a unitary transformative process of community pattern recognition for nurses and communities interested in understanding complex community dynamics and engaging in healthy transformations. PMID- 11890194 TI - An integrative model for environmental health research. AB - Environmental health research must achieve an integration of understanding, reaching from physiological research on health effects of toxic agents to actions that people may take, individually and collaboratively, to reduce their risks. This article proposes an integrative model of environmental health, encompassing four broad domains and their interrelationships: physiological, vulnerability, epistemological, and health protection. If we wish to empower communities to make the tough decisions necessary to truly protect the well-being of their most vulnerable members, each domain must be attended to, and links between scientific knowledge and social processes must be understood. PMID- 11890195 TI - Visible humans, vanishing bodies, and virtual nursing: complications of life, presence, place, and identity. AB - The emergence of the posthuman body and the disappearance of the humanist body serve as background for the rediscovery of the body as resource and problem in nursing. At the precise moment when the fleshy body is deemed increasingly irrelevant and immaterial in cyberspace come divergent moves in nursing toward not only resurrecting this body, but also toward virtual environments of nursing care, where fleshy bodies never encounter each other. The posthuman conflation of bodies and information poses the greatest challenge yet to the secure place, presence, and identity of nursing in health care. PMID- 11890196 TI - Creating a context of safety and achievement at school for children who are medically fragile/technology dependent. AB - Parents, nurses, and educators collaborate closely to create school environments that are safe and productive for children who are medically fragile/technology dependent. This article reports the results of a field study conducted in schools and family homes with the significant adults who care for and facilitate school participation for children who are medically fragile/technology dependent. Key steps in this process included learning about rights and responsibilities, planning for children's individual needs, being persistent in the face of systemic barriers, and taking actions to protect both children and professionals from perceived threats. PMID- 11890197 TI - Home and border: the contradictions of community. AB - If community is the theory and locale for much of nursing in the 21st century, complex thinking about community is required. In creating unified identities to form community, differences are suppressed and border communities are built. This article examines the contradictory, complicated, and unending processes of producing home (a space of comfort and commonality) and border (a space of discomfort and difference) communities. The construction of border communities in terms of class and race/ethnicity are considered through exemplars from research on the meaning of community. A deeper understanding of community calls for us to live with uncertainty and unfamiliarity. PMID- 11890198 TI - Living in a post-September 11 world. PMID- 11890199 TI - Mitochondrial creatine kinase binding to liposomes and vesicle aggregation: effect of cleavage by proteinase K. AB - Mitochondrial creatine kinase and its proteinase K nicked-derivative interaction with liposomes induced slight secondary structure changes evidenced by infrared spectra. In nondenaturing conditions, the N-terminal (K1) and the C-terminal (K2) fragments remained associated with each other and bound to liposomes. When the two fragments were separated by denaturation, K2 was soluble, whereas most of K1 was adsorbed onto liposomes. The three-dimensional structure of uncleaved mtCK suggests that the C-terminal moiety, which contains positively charged surface residues, interacted with membranes. After denaturation and renaturation of the nicked enzyme, both peptides did not refold properly and did not reassociate with each other. The misfolded K1 fragment bound to the membrane through a stretch of positive residues, which were buried in the native enzyme. The lack of binding of the ill-folded K2 peptide could be related to the disruption of the optimal disposition of its positive charges, responsible for the correct interaction of native mtCK with membrane. PMID- 11890200 TI - Interaction of cAMP receptor protein from Escherichia coli with cAMP and DNA studied by dynamic light scattering and time-resolved fluorescence anisotropy methods. AB - Cyclic AMP receptor protein (CRP) regulates the expression of more than 100 genes in Escherichia coli when complexed with cyclic AMP. Dynamic light scattering (DLS) and fluorescence decay anisotropy measurements of CRP were performed in solution, in the absence and presence of cAMP. We have also measured the effect of DNA sequences, including lac and gal promoter sequences, on the shape of CRP DNA complexes. DLS measurements show that upon cAMP binding at low nucleotide concentration, the Stokes radius decreases from the value of 2.8 nm for apo-CRP to the value of 2.7 nm. At higher cAMP concentration, only a very small further decrease was detected. Fluorescence anisotropy decay measurements, with the use of CRP labeled at Cys-178 with 1,5-I-AENS, indicate that apo-CRP exhibits two rotational correlation times. The longer time, theta1 = 23.3 ns, corresponds to the overall motion of the protein, and the shorter time, theta2 = 1.4 ns, exhibits segmental mobility of the C-terminal domain of CRP. Binding of cAMP into CRP induced substantial increase of theta1 to the value of 30.7 ns, whereas theta2 remained unchanged. The DLS measurements indicate that the binding of CRP into a fragment of DNA possessing a sequence of lac promoter induces a larger increase in the Stokes radius of lac-CRP complex than in case of gal-CRP complex. Similarly, a higher change was detected in rotational correlation time, theta1, in the case of lac-CRP complex than in case of gal-CRP. Because the lac and gal promoters are characteristic for the two different classes of CRP-dependent promoters, one can expect that the observed differences in lac-CRP and gal-CRP complexes are important in activation of transcription in Escherichia coli. PMID- 11890201 TI - Carboxyl-terminal modification alters the subunit interactions and assembly pathways of normal and sickle hemoglobins. AB - Parallel isofocusing studies established that carboxypeptidase A removal of the His-146 (HC3) and Tyr-145 (HC2) residues of beta heme subunits affected the assembly properties of both Des beta(A) and Des beta(S) with alpha heme chains, albeit to differing degrees. Indeed, the rate of Des beta(A) oligomer dissociation (k1), as determined by visible spectroscopy, was 4.3-fold faster than that of its native beta(A) counterpart. Furthermore, Soret spectral studies have affirmed distinct rates of normal (HbA), sickle (HbS), and Des HbA hemoglobin assembly (k'2) from their alpha and beta [Des beta(A)] heme-containing monomers. Matching kinetic analysis of Des beta(A) and Des beta(S) chain assembly (with an identical a chain) revealed 4.6- and 7.8-fold faster combination rates than those seen for beta(A) and beta(S) chains, respectively. This 3-fold disparity in rates strongly supports the critical role of the beta-6 (A3) residue, and its amino-terminal region, in a chain partner recognition and subsequent human hemoglobin assembly. PMID- 11890202 TI - Characterization of active barley alpha-amylase 1 expressed and secreted by Saccharomyces cerevisiae. AB - Recombinant barley alpha-amylase 1 isozyme was constitutively secreted by Saccharomyces cerevisiae. The enzyme was purified to homogeneity by ultrafiltration and affinity chromatography. The protein had a correct N-terminal sequence of His-Gln-Val-Leu-Phe-Gln-Gly-Phe-Asn-Trp, indicating that the signal peptide was efficiently processed. The purified alpha-amylase had an enzyme activity of 1.9 mmol maltose/mg protein/min, equivalent to that observed for the native seed enzyme. The kcat/Km was 2.7 x 10(2) mM(-1) x s(-1), consistent with those of alpha-amylases from plants and other sources. PMID- 11890203 TI - Purification and characterization of a new trypsin inhibitor from Dimorphandra mollis seeds. AB - A second trypsin inhibitor (DMTI-II) was purified from the seed of Dimorphandra mollis (Leguminosae-Mimosoideae) by ammonium sulfate precipitation (30-60%), gel filtration, and ion-exchange and affinity chromatography. A molecular weight of 23 kDa was estimated by gel filtration on a Superdex 75 column SDS-PAGE under reduced conditions showed that DMTI-II consisted of a single polypeptide chain, although isoelectric focusing revealed the presence of three isoforms. The dissociation constant of 1.7 x 10(-9) M with bovine trypsin indicated a high affinity between the inhibitor and this enzyme. The inhibitory activity was stable over a wide pH range and in the presence of DTT. The N-terminal sequence of DMTI-II showed a high degree of homology with other Kunitz-type inhibitors. PMID- 11890204 TI - Interactions between esterified whey proteins (alpha-lactalbumin and beta lactoglobulin) and DNA studied by differential spectroscopy. AB - Spectroscopic study of interactions between esterified whey proteins and nucleic acids, at neutral pH, showed positive differential spectra over a range of wavelength between 210 and 340 nm. In contrast, native forms of whey proteins added to DNA did not produce any differential spectra. The positive difference in UV absorption was observed after addition of amounts of proteins as low as 138 molar ratio (MR) of protein/DNA, indicating high sensitivity of the applied method to detect interactions between basic proteins and DNA. UV-absorption differences increased with MR of added whey protein up to saturation. The saturation points were reached at relatively lower MR in the case of methylated forms of the esterified protein as compared to its ethylated form. Saturation of nucleic acid (2996 bp long) was achieved using 850 and 1100 MR of methylated beta lactoglobulin and of methylated alpha-lactalbumin, respectively. Saturation with ethylated forms of the proteins was reached at MR of 3160 and 2750. Lysozyme, a native basic protein, showed a behavior similar to what was observed in the case of methylated forms of the dairy proteins studied. However, in the case of lysozyme, saturation was achieved at relatively lower MR (700). Methylated , casein failed to give positive spectra at pH 7 in the presence of DNA. It interacted with DNA only when the pH of the medium was lowered to 6.5, below its pI. Generally, amounts of proteins needed to saturate nucleic acid were much higher than those needed to neutralize it only electrostatically, demonstrating the presence on DNA of protein-binding sites other than the negative charges on the sugar-phosphate DNA backbones. Addition of 0.1% SDS to the medium suppressed totally all spectral differences between 210-340 nm. The presence of 5 M urea in the medium reduced only the spectral differences between 210-340 nm, pointing to the role played by hydrophobic interactions. Peptic hydrolysates of esterified and native proteins or their cationic fractions (pH > 7) produced negative differential spectra when mixed with DNA. The negative differences in UV absorption spectra were the most important in the case of peptic hydrolysates of methylated derivatives of whey proteins. PMID- 11890206 TI - Secondary structure prediction and folding of globular protein: refolding of ferredoxin. AB - The physicochemical mechanism of protein folding has been elucidated by the island model, describing a growth type of folding. The folding pathway is closely related with nucleation on the polypeptide chain and thus the formation of small local structures or secondary structures at the earliest stage of folding is essential to all following steps. The island model is applicable to any protein, but a high precision of secondary structure prediction is indispensable to folding simulation. The secondary structures formed at the earliest stage of folding are supposed to be of standard form, but they are usually deformed during the folding process, especially at the last stage, although the degree of deformation is different for each protein. Ferredoxin is an example of a protein having this property. According to X-ray investigation (1FDX), ferredoxin is not supposed to have secondary structures. However, if we assumed that in ferredoxin all the residues are in a coil state, we could not attain the correct structure similar to the native one. Further, we found that some parts of the chain are not flexible, suggesting the presence of secondary structures, in agreement with the recent PDB data (1DUR). Assuming standard secondary structures (alpha-helices and beta-strands) at the nonflexible parts at the early stage of folding, and deforming these at the final stage, a structure similar to the native one was obtained. Another peculiarity of ferredoxin is the absence of disulfide bonds, in spite of its having eight cysteines. The reason cysteines do not form disulfide bonds became clear by applying the lampshade criterion, but more importantly, the two groups of cysteines are ready to make iron complexes, respectively, at a rather later stage of folding. The reason for poor prediction accuracy of secondary structure with conventional methods is discussed. PMID- 11890205 TI - Conformational study of N(epsilon)-(carboxymethyl)lysine adducts of recombinant gammaC-crystallin. AB - N(epsilon)-(carboxymethyl)lysine, an advanced glycation end product, is present in the human lens. The effects of CML formation on protein conformation and stability were studied using the recombinant gammaC-crystallin as a model. Conformational change was studied by spectroscopic measurements such as fluorescence and circular dichroism. Conformational stability was determined by unfolding with heat. The results indicated that no conformational change was observed due to CML formation, but conformational stability decreased. These observations can be explained in terms of the relatively stable structure of gamma-crystallin, especially when compared with other crystallins. The lens nucleus is rich in gamma-crystallin and its stable conformation can assist gamma crystallin sustained insults and remain soluble. PMID- 11890207 TI - Mechanisms of liver injury relevant to pediatric hepatology. AB - Hepatocyte injury and necrosis from many causes may result in pediatric liver disease. Influenced by other cell types in the liver, by its unique vascular arrangements, by lobular zonation, and by contributory effects of sepsis, reactive oxygen species and disordered hepatic architecture, the hepatocyte is prone to injury from exogenous toxins, from inborn errors of metabolism, from hepatotrophic viruses, and from immune mechanisms. Experimental studies on cultured hepatocytes or animal models must be interpreted with caution. Having discussed general concepts, this review describes immune mechanisms of liver injury, as seen in autoimmune hepatitis, hepatitis B and C infection, the anticonvulsant hypersensitivity syndrome, and autoimmune polyendocrinopathy. Of the monogenic disorders causing significant liver injury in childhood, alpha-1 antitrypsin deficiency and Niemann-Pick C disease demonstrate the effect of endoplasmic or endosomal retention of macromolecules. Tyrosinemia illustrates how understanding the biochemical defect leads to understanding cell injury, extrahepatic porphyric effects, oncogenesis, pharmacological intervention, and possible stem cell therapy. Pathogenesis of cirrhosis in galactosemia remains incompletely understood. In hereditary fructose intolerance, phosphate sequestration causes ATP depletion. Recent information about mitochondrial disease, NASH, disorders of glycosylation, Wilson's disease, and the progressive familial intrahepatic cholestases is discussed. PMID- 11890208 TI - Clinical significance of cytokine determination in synovial fluid. AB - Cytokines are a complex family of small regulatory proteins able to mediate intercellular communication and play a crucial role in immunologic and inflammatory reactions. Many reports have demonstrated that some cytokines, in particular tumor necrosis factor alpha (TNFalpha) and interleukin (IL)-1beta, IL 6, and IL-8, so-called proinflammatory, may have a major role in the pathogenesis of joint diseases. Thus, high levels of these substances have been found in inflammatory arthropathies, in particular in those characterized by a more aggressive and destructive outcome, such as rheumatoid arthritis, gout, and infectious arthritis. In keeping with their role, the determination of cytokines in synovial fluid may be proposed for clinical purposes, including diagnostic and prognostic assessments. Furthermore, as some of these cytokines may reflect disease activity, their determination may also be useful in the evaluation of therapy. PMID- 11890209 TI - How to assess and counsel the older driver. AB - Suggesting that a patient stop driving is never easy, yet taking no action may result in deadly consequences. Open communication with the patient and the family in the office setting can help physicians assess risk for a driving accident. The physician can then decide if further assessment and perhaps rehabilitation will benefit the patient. Working with the family and involving community resources to secure alternative forms of transportation may also be needed. PMID- 11890210 TI - A 52-year-old man with excessive daytime sleepiness. PMID- 11890211 TI - Oncologic emergencies for the internist. AB - Most cancer patients experience at least one emergency during the course of the disease. This paper reviews the diagnosis and treatment of tumor lysis syndrome, hypercalcemia of malignancy, superior vena cava syndrome, spinal cord compression, strokes and seizures, and treatment-related emergencies. PMID- 11890212 TI - Hereditary hemochromatosis: a common, often unrecognized, genetic disease. AB - Hereditary hemochromatosis in people of northern European descent is more common than many physicians realize. It causes excessive gastrointestinal absorption of iron, leading to potentially fatal iron deposition in multiple organs. Early diagnosis and phlebotomy to reduce iron stores can prevent complications and provide normal life expectancy. Genetic testing of relatives of patients with hemochromatosis is warranted in some circumstances. PMID- 11890213 TI - Patient information. What is hemochromatosis? PMID- 11890214 TI - Hereditary hemochromatosis: molecular genetic testing issues for the clinician. PMID- 11890215 TI - The B-type natriuretic peptide assay: a rapid test for heart failure. AB - The B-type or brain natriuretic peptide (BNP) assay, a 15-minute bedside blood test, is highly sensitive and fairly specific for diagnosing heart failure and is useful in evaluating suspected heart failure in outpatients and in emergency care. Other uses include screening for left ventricular dysfunction and predicting outcome in patients with an established diagnosis of heart failure or myocardial infarction. PMID- 11890216 TI - How to use nesiritide in treating decompensated heart failure. AB - Nesiritide (Natrecor), a synthetic formulation of B-type natriuretic peptide (BNP), is the first new parenteral agent to be approved for treating heart failure in more than a decade. In patients hospitalized with decompensated congestive heart failure, nesiritide promptly reduces pulmonary capillary wedge pressure, pulmonary arterial pressure, right atrial pressure, and systemic vascular resistance, resulting in clinical improvement. PMID- 11890218 TI - What is the best way to determine if thrombocytopenia in a patient on multiple medications is drug-induced? PMID- 11890217 TI - What is the best diagnostic approach when pheochromocytoma is suspected? PMID- 11890219 TI - A girl with congenital heart defect, deformed hands and a sacral mass. PMID- 11890220 TI - Doppler echocardiographic evaluation of pulmonary artery pressure in children with acute pneumonia. AB - In this prospective study 37 children (ranging 2 months-15 years) with acute pneumonia were evaluated by Doppler echocardiography for the presence of pulmonary hypertension (PH). The goal of this study was to determine the frequency of PH in children with acute pneumonia because the diagnosis of PH influenced the treatment of pneumonia in these patients. The patients who had more than 35 mmHg of systolic pulmonary arterial pressure were considered to have PH. In our study PH was found in 15 (40.5%) of 37 patients. We did not find any significant difference for the parameters including the age, weight, height, clinical symptoms and signs (fever, cough, dyspnea, tachycardia and tachypnea etc.), and laboratory findings such as hemoglobin, PCO2, HCO3 and PO2 between the patients with and without PH (p>0.05). However, there was a significant difference in cyanosis, cardiac failure, blood pH level and O2 saturation measured by pulse oximetry between the patients with and without PH (p<0.05). PMID- 11890221 TI - Oscillometric ambulatory blood pressure values in healthy children. AB - To assess the feasibility of ambulatory blood pressure monitoring (ABPM) and establish reference values for ABPM in normal children and adolescents, 24-hour ABPM was performed in 120 normotensive, nonobese subjects, aged from 6 to 14 years (58 males, age 10.1 +/- 2.8 years; 62 females, age 9.7 +/- 3.1 years; mean +/- SD). The subjects were classified into three age groups: group A (6-8 years, n=40, 23 males, 17 females), group B (9-11 years, n=40, 21 males, 19 females), and group C (11-14 years, n=40, 18 males, 22 females). ABPM was carried out using an oscillometric device with appropriate cuff size. The monitor was programmed to measure BP every 15 min during the day (6 a.m. to 10 p.m.) and every 30 min during the night (10 p.m. to 6 a.m.). Successful rates (recording time=24 hours and > 40 recordings) were 83%, 88%, and 93% in groups A, B, and C, respectively. The recordings of 15 subjects (7 in group A, 5 in group B, 3 in group C) were not analyzed because of incomplete readings. The 24-hour mean systolic/diastolic blood pressure (SBP/DBP) for males and females were 106 +/- 7/65 +/- 5 and 105 +/ 6/64 +/- 5, 110 +/- 6/66 +/- 5 and 109 +/- 5/65 +/- 4, and 115 +/- 7/67 +/- 6 and 113 +/- 6/66 +/- 4 mmHg in groups A, B, and C, respectively. The daytime mean SBP/DBPfor males and females were 110 +/- 7/73 +/- 4 and 109 +/- 8/72 +/- 5, 117 +/- 5/75 +/- 4 and 115 +/- 8/74 +/- 6, and 122 +/- 6/76 +/- 4 and 120 +/- 7/75 +/ 6 mmHg in groups A, B, and C, respectively. The nighttime mean SBP/DBP for the corresponding sex and age groups were 97 +/- 8/58 +/- 5 and 95 +/- 7/57 +/- 6, 101 +/- 8/59 +/- 6 and 99 +/- 7/57 +/- 5, and 105 +/- 7/58 +/- 5 and 103 +/- 6/56 +/- 4 mmHg, respectively. The first fall in BP appeared after lunch (approximately 1:00 pm) and the second fall appeared during the night hours (11:00 pm to 6:00 am). There was a significant correlation between 24-hour mean ambulatory SBP and age (r=0.48, P < 0.001). In contrast, ambulatory DBP and age were not significantly correlated. A nocturnal fall of SBP/DBP was observed in all age- and sex-subgroups. This study showed that ABPM was reproducible and accurate. The normal ambulatory blood pressure (ABP) values in the pediatric age group have not been reported in Taiwan. The distribution of ABP observed in this study could serve as a preliminary reference. A multicenter study may be done to provide normal ranges of ABP in Taiwan for clinical purposes. Further studies are necessary to elucidate the role of ABPM in children. PMID- 11890222 TI - Placental weight and birth characteristics of healthy singleton newborns. AB - To determine the relationship of placental weight and birth characteristics of healthy Chinese singleton newborns, 552 consecutive singleton near-term and uncomplicated pregnancies were enrolled in a retrospective study from October 2000 to April 2001. Maternal age, infants' gender, gestational age, placental weight (PW), birth weight (BW), birth length (BL), cord length, ponderal index, and appearance of meconium-stained amniotic fluid or a nuchal cord were recorded and analyzed. Our results show that the mean value of BW was 3235 +/- 17 g, birth length was 48.8 +/- 0.1 cm, PW was 646.2 +/- 0.3 g, BW/BL was 66.2 +/- 0.3, BW/PW was 5.1 +/- 0.1, and ponderal index was 2.8 +/- 0.0, respectively. Male neonates had significantly larger BWs (p<0.05) and BLs (p<0.01) than did female neonates. However, there was no significant difference in PW between male and female neonates. PW was positively correlated with BW (r=0.413, p<0.01), BL (r=0.305, p<0.01), BW/BL (r=0.397, p<0.01), and cord length (r=0.264, p<0.01). The PW of studied infants showed no significant difference when grouped by gestational age, maternal age, presence of meconium stained amniotic fluid, or presence of a nuchal cord. Of the studied healthy neonate, 81.9% and 69.4% had no meconium stained amniotic fluid or nuchal cord, respectively. There were no significant differences in PW or BW/PW between neonates with the presence or absence of meconium-stained amniotic fluid or a nuchal cord. These results indicate that PW has a significant role in fetal growth in terms of weight, body length, and cord length. Furthermore, it has no significant role in the presence of meconium stained fluid or a nuchal cord in healthy neonates. PMID- 11890223 TI - Clinical aspects of X-linked hypophosphatemic rickets. AB - X-link hypophosphatemia (XLH) is the most common heritable form of rickets and is usually transmitted as an X-linked dominant disorder Until now there is no report about clinical data of XLH in Taiwan. We retrospectively studied 15 patients (5 males and 10 females) of XLH in our hospital in the past 18 years to delineate the clinical aspects of this disease. A total of 7 patients had family histories consistent with XLH; 8 appeared to be sporadic cases. Fourteen of 15 cases had typical physical and radiologic findings of rickets except case 1 who only had waddling gait and sacroiliitis with the initial complaint of joint pain. All 15 cases had normal serum creatinine, normocalcemia, normal 1,25-dihydroxyvitamin D (1,25-(OH)2D), reduced serum phosphate concentration, reduced urinary calcium excretion and reduced TmP/GFR. All but one had reduced %TRP. Thirteen of 15 cases had elevated alkaline phosphatase activity; two showed hyperparathyroidism. Four cases had sufficient follow-up growth data and had an increase in height percentile and velocity after combination therapy. PMID- 11890224 TI - The clinical efficacy of in vitro allergen-specific IgE antibody test in the diagnosis of allergic children with asthma. AB - Asthma is a very common respiratory allergic disease in Taiwan. The aims of this study were to investigate the allergen-sensitized profile and its relationship with serum total IgE levels in allergic asthmatic children in Taiwan. Moreover, the number of allergens to be tested for the most efficient and effective diagnosis of allergic diseases was also examined. Total IgE and IgE specific for a panel of common aeroallergens were assayed in 200 serum samples of asthmatic children using Pharmacia CAP system (Pharmacia, Uppsala, Sweden). House dust mites Der p (Dermatophagoides pteronyssinus), Der f (Dermatophagoides farinae), and Bt (Blomia Tropicalis) had the highest sensitized rates at 82.5%, 82.0%, and 72%, respectively. Candida albican (14.0%) and Bermuda grass (8.0%) were the most common sensitized fungus and pollen in our subjects, respectively. The accumulated sensitized rate (10%) for pollens was lower than those of fungus (21.5%) and house dust allergens (84%). The average serum total IgE of the allergen-negative asthmatic children (n=30) was significantly lower than that of children with at least one allergen sensitized asthma (n=170) (377.9 +/- 123.6 vs. 1117.8 +/- 235.7 IU/ml, p<0.05). The level of total IgE was significantly correlated with the concentrations of mite-specific IgE antibodies, but not with the numbers of allergen sensitized. In addition, the detection rate was 84% when the 4 most common allergens (Der p, Der f, Dog dander, and cockroach) were tested, similar to the result after testing for all 12. PMID- 11890225 TI - Human DNA repair defects: are they predisposing to cancer? PMID- 11890226 TI - Measurement of ear length in neonates, infants and preschool children in Taiwan. AB - Ear length is important in the evaluation of congenital anomaly syndrome, such as small ear in Down syndrome. In this research, 4,416 normal Chinese children in Taiwan were enrolled into our study. The sample of 284 full term neonates, 2,742 infants and children aged from one month to three years, and 1,420 preschool children were measured for ear length. We placed a ruler along the greatest vertical axis of the right ear We calculated the mean value and standard deviation of the ear length in normal Chinese neonates, infants and preschool children in Taiwan. No significant sex differences were observed. Twenty patients with Down syndrome were also measured. Compared with the previous study, there was no significant difference between the ear length of Chinese children and that of Caucasian children. The ear length of patients with Down syndrome was indeed smaller than that of the normal children. Due to the correlation of reduced ear length and congenital anomaly, we should set up our own data about the growth of the ear length. Thus we can provide an additional clue to diagnose the congenital anomaly syndromes. PMID- 11890228 TI - Cockayne syndrome in a family. AB - Cockayne syndrome is one of the families of rare progeroid syndromes. We report on two female siblings suffering from Cockayne syndrome. At birth, they both appeared normal, although both demonstrated a low birth weight and breech presentation. The first-born child died at the age of eight months with associated contracted limbs, brain calcification, and photosensitivity. The younger sibling exhibited short stature, microcephaly, a beaked nose and malformed ears, spasticity, photosensitivity, pigmented degeneration of the retina, and psychomotor retardation at the age of six years. Intracranial calcification and the absence of a brain stem-evoked potential were also noted. Testing her skin fibroblasts, which showed a moderate UV sensitivity and a severe deficiency of transcription-coupled repair established the diagnosis of Cockayne syndrome. Genetic counseling was offered for the family. PMID- 11890227 TI - Mesenteric lymphangioma causing bowel obstruction: report of one case. AB - A 6-year-old female was sent to our ER due to nausea, vomiting and abdominal distension for 2 days. This child had a history of constipation and failed intermittent medical treatment for 2 years. Her plain abdominal X-ray showed multiple intestinal loops and under the impression of acute abdomen with mechanical intestinal obstruction, an exploratory laparotomy was performed. A huge mesenteric tumor was discovered to be the cause of the intestinal obstruction; the involved bowel and the mesenteric lymphangioma were resected and primary anastomosis was done. Mesenteric cystic lymphangioma is a rare cause of bowel obstruction; preoperative diagnosis is difficult due to silent clinical course and lack of awareness of the clinical and morphological features of this disease. The case is presented along with a review of literature with the conclusion that a high index of suspicion is recommended. An abdominal ultrasonography may be recommended to evaluate a long-term constipated child to ascertain that any cystic lesion will not be missed. PMID- 11890229 TI - Neonatal solitary liver abscess: report of one case. AB - We report a case of a solitary liver abscess in a 5-week-old female. She was full term, and there were no predisposing events or immune deficiencies. The only sign of her disease was a gradually distended abdomen. A prior episode of fever with possible occult bacteremia was implicated in the development of her abscess. The abdominal sonography and magnetic resonance image (MRI) did not provide any definite preoperative diagnostic information. Surgical resection of the abscess and a short course of antibiotic therapy cured the disease. This patient was still well following 2 years of check-ups by sonography. The possibility of a pyogenic liver abscess should be considered in the differential diagnosis of neonatal hepatic mass. That is, even if there is not a definite diagnostic focus on finding an infection. PMID- 11890230 TI - What's new in enuresis? AB - Treatment of primary nocturnal enuresis (PNE) using desmopressin (dDAVP) is based upon the hypothesis that antidiuretic hormone (arginine vasopressin (AVP) secretion is insufficient during the night. Persisting doubts about the theoretical background of this treatment gave reason to different studies: In a first study, AVP regulation in children with PNE was investigated. AVP levels after fluid restriction were compared to those of healthy controls. In order to maintain osmolality, the plasma AVP concentrations of the enuretic children rose to significant higher levels than in the controls. In a second study, the short time memory of children treated with dDAVP because of PNE was measured. Children under dDAVP showed a significantly better short- time memory. CONCLUSION: The results are consistent with the established fact that AVP secretion is a function of plasma osmolality. They contradict the hypothesis that enuretic children have a AVP deficiency which has to be supplemented. Rather, the results point to central action of dDAVP, a defect at the central AVP receptor level or the signal transduction pathway. PMID- 11890231 TI - Surgical skills assessment: an ongoing debate. PMID- 11890232 TI - Simulation in urology--a role for virtual reality? PMID- 11890233 TI - Spatial awareness in urologists: are they different? AB - OBJECTIVE: To compare innate spatial awareness skills, using the MIST-VR system (Ethicon Ltd, Edinburgh, a computer-based virtual reality system that objectively tests spatial awareness) among three groups of people (consultant urologists, urological trainees and controls who were not surgeons), because urological surgeons require spatial awareness for endoscopic and laparoscopic surgery, but trainees are selected by academic prowess rather than surgical aptitude. SUBJECTS AND METHODS: The MIST-VR system was used to test 122 volunteers in three groups, i.e. 39 consultant urologists, 46 urological trainees and 37 controls (not surgeons). The demographic data recorded for each group included age, sex, eyesight, handedness, and endoscopic and laparoscopic experience. Volunteers performed a repetitive series of three tasks using the system. Their performance was measured in terms of time, errors and economy of movement, as well as the duration and accuracy of diathermy in Task 3. RESULTS: The consultants were significantly older than the trainees and controls (both P<0.001) and had more endoscopic experience (P=0.005). In Task 1, the trainees made significantly fewer errors (P=0.045) and had a greater economy of movement (P=0.03) than the controls. In Task 2 the trainees performed the task more rapidly than the consultants (P=0.04) and controls (P=0.02). Trainees were more economical in movement than were consultants (P=0.031) and controls (P=0.046). In the more complex Task 3, trainees outperformed consultants in terms of errors (P=0.03), economy of movement (P=0.046), total diathermy time (P=0.005) and diathermy error (P=0.03). Controls performed similarly to the consultants. Although there was a trend towards better performance by trainees over controls, this was only significant for time (P=0.04) and total diathermy time (P=0.011). A few participants had results that were >2 SD above the mean and several people could not complete Task 3. CONCLUSIONS: Urologists do not differ from the general population in terms of innate spatial ability in this setting. There are several people who may have a defect in spatial awareness but the incidence was the same in each group. Urological trainees outperformed consultants in these tasks; the reasons for this are unclear. The MIST-VR system is of no help in aptitude testing for urological trainees, although it may have a role in teaching laparoscopic surgery. Testing other psychometric components may be more important for acquiring surgical skills than innate spatial-awareness skills. Further studies are required to investigate this possibility. PMID- 11890234 TI - Renal cell carcinoma: incidental detection during routine ultrasonography in men presenting with lower urinary tract symptoms. AB - OBJECTIVE: To compare renal cell carcinomas (RCCs) presenting incidentally in patients referred for lower urinary tract symptoms (LtJTS) with those presenting symptomatically, by stage, intervention and outcome. PATIENTS AND METHODS: The case notes of all male patients (100) diagnosed with RCC between 1991 and 1998 were reviewed and modes of presentation recorded. The patients were divided into two groups: those who were referred with LUTS (frequency, urgency, hesitancy, poor stream, nocturia) and in whom RCC would not have been suspected and was thus detected incidentally on routine ultrasonography; and all patients in whom carcinoma might have been suspected from their symptoms but, for the purposes of this study, also included patients in whom RCC was diagnosed during ultrasonography for unrelated intra-abdominal pathology. Details of diagnostic imaging and clinical staging were similarly recorded for both groups and where surgical intervention was undertaken, histopathological data were also noted. The clinical course and long-term outcome of incidentally detected tumours was then compared with their symptomatically presenting counterparts. RESULTS: The mean (range) follow-up for all patients was 30 (1.5-96) months; for those in the incidental group it was 31 (1-86) months and in the symptomatic patients 29 (1 96) months. Organ-confined disease was found in two-thirds of patients with incidental tumours and in 38% of those in whom the tumour may have been suspected; the difference was statistically significant (chi-squared test P<0.05). The mean (SD) size of tumours discovered incidentally and in symptomatic patients was 5.9 (1.94) cm and 9.2 (3.39) cm, respectively; this difference was also statistically significant (t-test, P<0.001). Of the 24 patients with incidentally detected tumours, 14 (58%) were alive with no recurrence, and of the 76 presenting symptomatically, 27 (35%) were alive with no recurrence at the last follow-up; disease survival curves showed a statistically better survival rate for those with organ-confined tumours. CONCLUSION: Incidentally diagnosed RCC represents a significant proportion of those who are ultimately diagnosed with the malignancy. Opportunities which arise for appropriate screening of the upper tracts during routine urological investigations (e.g. ultrasonography of the upper tracts in patients referred for LUTS) should be endorsed, contrasting with the more traditional approach, which argues that it yields no ultimate survival advantage. PMID- 11890235 TI - Steinstrasse after extracorporeal shockwave lithotripsy: aetiology, prevention and management. AB - OBJECTIVE: To determine the causes of steinstrasse, methods of prevention and treatment strategies. PATIENTS AND METHODS: Fifty-two patients with steinstrasse were identified and treated: all patients were initially treated conservatively but when there was obstruction, infection or no progression of the stone fragments, further treatment was used, ranging from repeated extracorporeal shockwave lithotripsy (ESWL), percutaneous nephrostomy (PCN), endoscopic manipulations and finally open surgery, depending on the degree of obstruction, infection, renal function and response to each kind of therapy. RESULTS: Conservative management was successful in 25 patients (48%), repeated ESWL in 12 (23%), PCN in 10 (19%), ureteroscopy in three (6%) and open surgery in two (4%). CONCLUSION: As many patients, and particularly those with larger stones, are treated by ESWL, the risk of developing steinstrasse will increase, with associated patient discomfort, infection or impaired renal function. The optimum selection of cases (aiming to pulverize the stones rather than fragment them) and accurate stone targeting are essential to minimise the development of steinstrasse. The meticulous follow-up of patients with steinstrasse should prevent any loss of renal function. When there is obstruction and/or infection or renal damage, active treatment is indicated, of which ESWL and PCN are the most effective, with ureteroscopy and open surgery reserved for difficult cases. PMID- 11890236 TI - Upper and mid-ureteric stones: a prospective unrandomized comparison of retroperitoneoscopic and open ureterolithotomy. AB - OBJECTIVES: To review our experience of retroperitoneoscopic ureterolithotomy (RPUL) and to compare the results with those from open surgery. PATIENTS AND METHODS: Between March 1994 and mid-December 2000, 55 patients with large (mean 2.1 cm) upper and mid-ureteric calculi, and with normal renal values, underwent RPUL. In 22 patients, earlier attempts with extracorporeal shock wave lithotripsy and ureteroscopy had failed. These patients were compared with 26 (mean stone size 2.4 cm) who underwent open ureterolithotomy during the same period. The two groups had similar distributions for age, sex, stone size and stone location; most stones were calcium-based. RESULTS: The mean operative duration and blood loss for RPUL and open surgery were 108.8 and 98.8 min, and 58.5 and 50.5 mL, respectively (not significant). The mean analgesic (pethidine) requirement and hospital stay for RPUL and open surgery were 41.1 and 96.9 mg, and 3.3 and 4.8 days, respectively (P<0.001). The duration of convalescence was significantly less after RPUL than open surgery (1.8 weeks vs 3.1). There were 10 conversions, which occurred early in the series, and one significant complication amongst patients who underwent RPUL. CONCLUSIONS: RPUL is comparable with open surgery for operative duration and blood loss, but the laparoscopic procedure has significant advantages over open surgery for analgesia, hospital stay, recuperation and cosmesis. RPUL is a viable alternative for large upper and mid ureteric calculi and in those patients where a previous attempt at endourological management has failed. However, the technique requires significant training and experience before good results can be obtained. PMID- 11890237 TI - Does cystoscopy correlate with the histology of recurrent papillary tumours of the bladder? AB - OBJECTIVE: To correlate the cystoscopic appearance of recurrent papillary bladder tumours with the histology after transurethral resection, and thus ascertain whether cystoscopy can reliably identify low-grade, noninvasive papillary tumours suitable for outpatient fulguration. PATIENTS AND METHODS: In all, 150 recurrent papillary tumours of the bladder identified at outpatient flexible cystoscopy were classified as either low-grade and noninvasive (TaG1), high-grade and noninvasive (TaG3), or invasive (TIG3) tumours, and correlated with urine cytology and histology of tumour stage and tumour grade after transurethral resection. RESULTS: Cystoscopy classified 84 of the 150 papillary tumours as TaG1 and 66 as either TaG3 or T1G3. Cystoscopy correctly predicted the histology of 78 of 84 (93%) TaG1 tumours, 71 of 72 (98%) TaG1 tumours associated with a negative urine cytology, and 92% of TaG3 or T1G3 tumours. CONCLUSIONS: A skilled urologist can identify noninvasive, low-grade recurrent papillary bladder tumours on follow up cystoscopy that do not require biopsy and that may be treated by outpatient fulguration alone. PMID- 11890238 TI - 99mTechnetium-C595 radioimmunoscintigraphy: a potential staging tool for bladder cancer. AB - OBJECTIVES: To assess whether immunoscintigraphy using a conjugate of the anti MUC1 monoclonal antibody C595 and 99mTc could be used to target transitional cell bladder cancer after intravenous administration to patients. PATIENTS AND METHODS: Twenty-one patients with invasive or metastatic transitional cell carcinoma were recruited. Patients received 1 mg of C595 labelled with 800 MBq 99mTc followed by imaging at 0.5, 6 and 24 h using a combination of planar and single-photon emission computed tomography. Of these patients, 14 subsequently underwent cystectomy, four underwent radiotherapy and the remaining three had histologically confirmed metastatic disease. The results of immunoscintigraphy were compared with surgical findings and conventional radiology. RESULTS: There were no adverse reactions in any patient. Of the 20 patients who were found to have tumour at the time of the study, positive localization of antibody in tumour was apparent in 16. Of the remaining four patients, false-positive localization of antibody in presumed nodal tissue was detected in two. The remaining scan results were equivocal. In three patients, histologically confirmed pelvic nodal metastases that had not been detected on preoperative computed tomography were identified. CONCLUSION: These early results show the potential of 99mTc-C595 immunoscintigraphy for staging bladder cancer. A larger study is needed to fully evaluate the technique. PMID- 11890239 TI - A comparison of urinary nuclear matrix protein-22 and bladder tumour antigen tests with voided urinary cytology in detecting and following bladder cancer: the prognostic value of false-positive results. AB - OBJECTIVES: To evaluate the diagnostic and prognostic value of the nuclear matrix protein-22 (NMP22) and bladder tumour antigen (BTAstat) tests compared with voided urinary cytology (VUC) in detecting and following bladder cancer, assessing particularly the prognostic value of false-positive test results in patients followed up for bladder cancer. PATIENTS AND METHODS: From 739 patients suspected of having bladder cancer, voided urine samples for the NMP22 and BTAstat tests, and for VUC and urine analysis, were collected before cystoscopy. All patients underwent transurethral resection of bladder lesions or mapping. and were followed for a mean (range) of 27.3 (3-65) months. RESULTS: In the 406 patients with bladder cancer, the overall sensitivity was 85% for NMP22, 70% for BTAstat and 62% for VUC. For histological grades 1-3 the sensitivity in detecting transitional cell carcinoma was 82%, 89% and 94% for NMP22, 53%, 76% and 90% for BTAstat, and 38%, 68% and 90% for VUC, respectively. Although the sensitivity in detecting invasive carcinoma was >85% for all the tests. NMP22 and BTAstat were statistically more sensitive than VUC for superficial tumours. The optimal threshold value for NMP22, calculated using the receiver operating characteristics curve was 8.25 U/mL. The specificity was 68% for NMP22, 67% for BTAstat, and 96% for VUC. The specificity of VUC remained >87% and was independent of benign histological findings. In contrast, in patients with no apparent genitourinary disease on histology, NMP22 and BTAstat had significantly higher specificity (94% and 92%, respectively: P=0.003) than in the group with chronic cystitis (52% for both tests). Forty patients having no bladder cancer at biopsy had a recurrence after a mean (range) follow-up of 7.7 (3-15) months: all had a previous history of bladder cancer. According to subsequent recurrence, the prognostic positive and negative predictive values were 18% and 91% for NMP22, 13% and 88% for BTAstat, and 79% and 91% for VUC. Both false-positive VUC and NMP22 tests predicted recurrence (log-rank test, P<0.001 and P=0.004, respectively), but the BTAstat test produced no similar correlation (P=0.778). CONCLUSION: The NMP22 and BTAstat tests are better than VUC for detecting superficial and low-grade bladder cancer but they have significantly lower specificity. After excluding diseases with the potential to interfere in these tests the overall specificity of both tests is increased considerably. False positive results from NMP22 and VUC but not from BTAstat in patients followed up for bladder cancer correlate with future recurrences. PMID- 11890240 TI - The ability of the American Joint Committee on Cancer Staging system to predict progression-free survival after radical prostatectomy. AB - OBJECTIVE: To examine, in a retrospective analysis of outcome based on prostate specific antigen (PSA) levels, whether the 1992 and revised 1997 staging criteria for prostate cancer can be used to predict progression-free survival for patients after radical prostatectomy for pT2 and pT3 prostate cancer. PATIENTS AND METHODS: In all, 291 patients with a PSA determination during a 6-month interval after radical prostatectomy were analysed (mean follow-up 5.2 years). In the absence of a uniform system of pathological staging, the histopathological stage was defined according to the 1992 and 1997 American Joint Cancer Committee/Union Internationale Contre le Cancer (AJCC/UICC) tumour-nodes-metastases (TNM) staging classification. Findings were correlated with the PSA value after surgery. The subgroups of pT2 and pT3 disease were compared for the time to PSA progression, using Kaplan-Meier data analysis and the log-rank test. RESULTS: The biochemical progression-free 5-year survival rates for stage pT2 were 83% (pT2a), 81% (pT2b) and 62% (pT2c); there were no significant differences in the pT2 subgroups. The recurrence-free rates for pT3 were 79% (pT3a), 65% (pT3b) and 50% (pT3c); the actuarial recurrence-free rate was significantly different for patients with 1997 AJCC pT3a vs pT3b disease (P=0.0132). There was no significant difference in the 1992 AJCC stages pT2a vs pT2b (P=0.1232) and the recurrence-free rate was not significantly different for patients with 1992 AJCC pT3a vs pT3b disease (P=0.9). There was a significant difference in the likelihood of a PSA relapse between patients with positive and negative surgical margins (P=0.131). CONCLUSION: These results support the current revised 1997 AJCC/UICC staging system for prostate cancer. There is an urgent need to develop a pathological equivalent to the AJCC/UIC TNM clinical staging system. Greater clinical input and evaluation from different institutions are essential to reach consensus on pathological staging categories that maximize the predictability of outcome after definitive therapy. Crucial issues are the definition and quantification of extraprostatic extension and definition of surgical margin categories. PMID- 11890241 TI - A randomized trial of the choice of treatment in prostate cancer: design and baseline characteristics. AB - OBJECTIVE: To assess the effects of different approaches to decision-making on the treatment chosen for prostate cancer and on the patients' quality of life in prostate cancer. PATIENTS AND METHODS: A multicentre randomized trial was conducted, including all histologically confirmed cases of prostate cancer diagnosed between September 1993 and November 1994 in four Finnish hospitals. In the intervention group, the role of the patient in the choice of treatment was actively emphasized. In the control group, the treatment was chosen using standardized treatment protocols. The first intermediate endpoint was the patient's participation in decision-making and the next will be the treatment chosen in the intervention and control groups. The main outcome will be the quality of life. Clinical data on prognostic factors including age, tumour grade, stage, functional status and serum prostate-specific antigen (PSA) concentration was collected for comparison between the arms, and between those enrolled or not. RESULTS: In all, 210 of 251 eligible patients were randomized into the two arms. Patients were randomized before obtaining informed consent, which led to four patients already randomized refusing to participate. The 41 patients not enrolled were of similar age and grade distribution, but more frequently had extensive disease than had those enrolled in the trial. Three patients were unable to participate because of rapid deterioration in their general condition after randomization. There were no clear differences in baseline characteristics (including age, functional status, tumour grade and stage) of the patients between the arms. The distribution of PSA level differed slightly between the arms, which may require adjustment in the analyses. Patients in the intervention arm participated in decision-making more actively than those in the control arm. CONCLUSION: Randomized studies on ethical issues such as the patient's role in choosing treatment are feasible and likely to provide important information. PMID- 11890242 TI - The value of multiple core biopsies for predicting the Gleason score of prostate cancer. AB - OBJECTIVE: To evaluate the accuracy of Gleason grading of prostate cancer in multiple core biopsies, compared with the final Gleason score of total prostatectomy specimens, and to investigate whether the prediction of the correct Gleason score is improved by increasing the number of biopsies. PATIENTS AND METHODS: Before total prostatectomy, 121 men had a mean (range) of 10.0 (8-14) transrectal ultrasonography (TRUS)-guided core biopsies taken from the apex, mid medial, mid-lateral and basal regions, from the transition zone and from lesions detected on TRUS. The biopsies and prostatectomy specimens were reviewed and the Gleason scores assessed. RESULTS: The preoperative biopsies predicted the prostatectomy Gleason score exactly in 45.5% of the patients and within one Gleason score in 93.4%. The biopsies under-graded the prostate cancer in 38.8% and overgraded it in 15.7%. The weighted kappa value for exact agreement was 0.502. If one biopsy was positive for cancer, the prostatectomy Gleason score was predicted correctly in 43.8% and within one score in 93.8%, compared with 53.8% and 92.3%, respectively, if cancer was found in at least seven biopsies. If the mid-lateral and transition zone biopsies had been excluded from the biopsy protocol, 5% of the cancers would have been undetected. Among the remaining 115 cancers, grading accuracy only improved from 43.5% to 45.2% by adding biopsies to the sextant protocol. CONCLUSION: Despite a statistically significant agreement between biopsy and prostatectomy Gleason score, under-grading remains a major problem. The prediction of the prostatectomy Gleason score is only marginally improved by increasing the number of biopsies. PMID- 11890243 TI - Prediction of tumour volume and pathological stage in radical prostatectomy specimens is not improved by taking more prostate needle-biopsy cores. AB - OBJECTIVE: To determine what, if any, additional prognostic information is available from the prostate needle biopsy by comparing the number of biopsy cores obtained with the pathology assessed from the radical retropubic prostatectomy (RRP) specimen. PATIENTS AND METHODS: The results from 135 consecutive patients who underwent RRP at a single institution were reviewed. Needle biopsy information (number of cores, percentage of positive cores, laterality of the positive cores, and Gleason sum) were compared with the pathological data of the RRP specimen, including stage, Gleason sum and tumour volume. Patients were further stratified into those with six or fewer cores (96 men) or more than six cores (39 men). Clinical data, including biopsy information and pathological findings, were compared using univariate and multivariate models. RESULTS: Overall, univariate analysis showed that the total prostate-specific antigen (PSA) level, number of positive cores, bilateral positive cores and percentage of positive cores were directly correlated with tumour volume (P=0.01). Also, PSA and percentage of positive cores were directly correlated with extracapsular extension (P=0.008 and P=0.01, respectively). In the multivariate model, the most important independent predictors of RRP tumour volume and pathological stage were the preoperative PSA level and percentage of cancer in the biopsy (P<0.01). There was no significant relationship between the number of cores obtained and the predicted pathology of the RRP specimen. There were no differences in the number of positive cores, bilateral positive cores or percentage tumour in the cores between men with more or less than six biopsies. In men with more than six core biopsies, there was no significant increase in prognostic information for tumour volume and extracapsular extension, or a correlation between the Gleason sum on biopsy and the RRP specimen. Taking more than six biopsies did not result in a significantly greater detection of potentially indolent tumours (defined as a tumour volume of <0.5 mL). CONCLUSIONS: While taking more prostate needle biopsy cores seems to improve the detection of prostate cancer, there appears to be no major improvement in prognostic information over that gained from traditional sextant biopsies. Furthermore, the results suggest that the percentage of positive cores is the best predictor of both pathological stage and tumour volume, from among the information readily available from prostate needle biopsy. Given the variability in the number of cores obtained for diagnosis in clinical practice, these results add credence to the use of the percentage of positive cores in the biopsy set, with known predictors such as PSA and Gleason score, into future models that attempt to predict tumour biology. PMID- 11890244 TI - The prevalence of Peyronie's disease: results of a large survey. AB - OBJECTIVES: To determine the prevalence of Peyronie's disease, a localized connective tissue disorder of the penile tunica albuginea, the symptoms of which include palpable plaque, painful erections and curvature of the penis, in a large sample of men in Germany. SUBJECTS AND METHODS: A standardized questionnaire was sent to 8000 male inhabitants (age range 30-80 years) of the greater Cologne area (approximately 1.5 million inhabitants). Three questions about the self-diagnosis of Peyronie's disease were previously assessed for validity on 158 healthy men and 24 patients with confirmed Peyronie's disease. To optimize the response rate, the questionnaire was mailed three times to all the men. RESULTS: The response rate after the third mailing was 55.4% (4432 men): 142 men (3.2%, mean age 57.4 years, SD 13.4) reported the new appearance of a palpable plaque which, from the previous validation, was the most sensitive question and the main symptom of the disease. In men aged 30-39 years only 1.5% reported localized penile induration, compared with 3.0% in those 40-49 and 50-59 years, 4.0% in those 60-69 years and 6.5% of those > 70 years old. Newly occurring angulation was reported by 119 of the 142 men (84%) and painful erection by 66 (46.5%). The combination of the three symptoms (plaque, deviation and painful erection) was reported by 46 of the 4432 respondents (1.04%), i.e. 32% of the 142 men with penile induration; 58 of the 142 men (41%) reported erectile dysfunction. CONCLUSIONS: This is the first large cross-sectional, community-based study to examine the prevalence of Peyronie's disease. Using previously validated questions the prevalence of Peyronie's disease in the sample was 3.2%; this is much higher than indicated in previous reports. A comparably high prevalence is reported for diabetes and urolithiasis, suggesting that this 'rare' disease is more widespread than previously thought. PMID- 11890245 TI - The penile disassembly technique in the surgical treatment of Peyronie's disease. AB - OBJECTIVE: To present an approach for treating Peyronie's disease, using the penile disassembly technique for reconstructive surgery. PATIENTS AND METHODS: From November 1996 to September 2000, 74 patients with Peyronie's disease were treated surgically. The penile disassembly technique was used in 46 of the patients (mean age 51 years, range 21-63). The indications were severe penile deviation under the glans cap, plaque in the distal third of the corpora cavernosa with the 'hour-glass' phenomenon, and more than one plaque at different sites. The corporal bodies are separated from the glans, neurovascular bundle and urethra. The technique enables the complete preservation of all structures of the neurovascular bundle, especially if it is incorporated into the plaque. The method provides an excellent approach to the repair of all deformities on the completely free corpora cavernosa and that are affected by the plaque. In the plaque region, incisional grafts are placed using full-thickness penile skin or saphenous vein. The technique also enables reduction corporoplasty, i.e. amputation of the tips of the corpora cavernosa that include plaque, in those with sufficient penile length. Penile re-assembly involves joining the glans, neurovascular bundle, urethra and repaired corpora cavernosa into their normal anatomical relationships. RESULTS: The mean (range) follow-up was 27 (6-53) months. The penis was completely straightened in 40 patients (87%) but the deviation recurred in six. In four patients the deformity was <10 degrees and in two was <20 degrees. Penile shortening occurred in 9% of the patients. There was no evidence of inflammation or infection after surgery. There were no injuries of either the neurovascular bundle or urethra. CONCLUSION: The penile disassembly technique could be a good alternative to other surgical techniques in treating selected patients with Peyronie's disease; it allows an excellent approach to penile deformities which can then be easily and safely corrected. PMID- 11890246 TI - Enterocystoplasty. AB - Despite the problems of augmentation cystoplasty, on balance it has been a much better form of management of the lower urinary tract in patients with bladder neuropathy or high pressure detrusor contractions than the alternatives of rectal diversion, indwelling catheter or external urinary diversion. The metabolic consequences do not seem to interfere with general health in the medium term. The risk of perforation appears to be present with other forms of augmentation cystoplasty or bladder replacement. However, the results are far from perfect and the ideal technique will be one that: removes the need for intraperitoneal surgery and prevents the risk of intestinal adhesions; stops the development of intestinal mucus and stone formation; prevents the metabolic complications and potential bony complications during adolescence; at the same time improves the patient's resistance to UTI; maintains the same degree of long-term, good, low pressure urine storage and the consequent improvement and stability of the upper urinary tract. PMID- 11890247 TI - Ureterocystoplasty: the latest developments. PMID- 11890248 TI - Seromuscular colocystoplasty. PMID- 11890249 TI - Autoaugmentation gastrocystoplasty. PMID- 11890250 TI - Experience with demucosalized ileum for bladder augmentation. AB - OBJECTIVE: To assess the use of demucosalized ileum for bladder augmentation, following the same principles previously used with the sigmoid colon. PATIENTS AND METHODS: Eleven patients with bladder exstrophy or a neurogenic bladder had their bladders augmented with demucosalized ileum instead of the sigmoid colon, but following the same technique. The use of a Foley catheter with an inflated balloon greatly facilitated the dissection of the mucosa from the muscle. A silicone model, inserted in the bladder, was used to avoid shrinkage of the patch. An animal model was also used for total bladder replacement following the same principle. RESULTS: The mean (range) follow-up was 15.4 (2-25) months. There was a significant increase in bladder compliance in all patients. A bladder of good shape and compliance was obtained in the animal model, with epithelial growth detected in all cases. CONCLUSIONS: Demucosalized ileum can be used safely for bladder augmentation in the same way as with the sigmoid colon. The distension of the isolated patch greatly facilitates dissection between the mucosa and muscle. The animal model supported this method of bladder replacement. PMID- 11890251 TI - Bladder regeneration by tissue engineering. PMID- 11890252 TI - The effects of a selective noradrenaline reuptake inhibitor on the urethra: an in vitro and in vivo study. AB - OBJECTIVE: To identify the effects of a selective noradrenaline reuptake inhibitor (venlafaxine) on urethral perfusion pressure (UPP) in rabbits and rats, and thus assess its therapeutic potential for treating stress urinary incontinence. MATERIALS AND METHODS: Strips of bladder and proximal urethra were prepared from female New Zealand White rabbits. Each strip was electrically stimulated and the contractile responses of controls strips compared with those after pretreatment with venlafaxine (100 micromol/L). In separate experiments using 80 adult female Sprague-Dawley rats (250-300 g), changes in intravesical pressure and UPP after the intra-arterial and intra-urethral administration of phenylephrine, phentolamine, fluoxetine and venlafaxine were monitored using double-lumen catheters. RESULTS: Pretreatment with venlafaxine significantly decreased the contraction of bladder strips (P=0.01) and significantly increased the contraction of urethral strips (P=0.008). In vivo, phenylephrine administered by both routes significantly increased UPP (P=0.02); phentolamine (arterial) significantly decreased UPP (P=0.001); fluoxetine (arterial) had no effect on UPP, and venlafaxine (both routes) significantly increased UPP (both P<0.001). The intravesical pressure was not changed significantly in any animal. CONCLUSIONS: Venlafaxine effectively increased UPP both in vitro and in vivo; these results imply that venlafaxine may be useful for treating stress urinary incontinence, by increasing the UPP. PMID- 11890253 TI - Homotypic cell-cell contacts stimulate the motile activity of rat prostate cancer cells. AB - OBJECTIVE: To characterize the effect of homotypic cell-to-cell collisions upon the motile activities of two rat prostatic cancer cell lines of markedly different metastatic potential. MATERIALS AND METHODS: The movements of strongly and weakly metastatic MAT-LyLu and AT-2 cells, respectively, were recorded under an inverted microscope at 37 degrees C. The motile activities of the cells at various cell densities were characterized quantitatively by computer-aided tracking methods and image analysis. The following variables were assessed: speed of movement, final displacement, coefficient of movement efficiency, diffusion constant and positive flow. RESULTS: MAT-LyLu and AT-2 cells showed only limited motile activity in sparse cultures where there was little contact amongst the cells. However, under these and all other subsequent conditions tested, the motile activity of the MAT-LyLu cells was higher than the AT-2 cells. As the density of the cultured cells was increased (leading to more cell-to-cell contacts) there was a significant increase in motility. This effect was more pronounced for the AT-2 than for the MAT-LyLu cells, resulting in visible acceleration of movement by direct physical contact among the colliding cells. The motile activities of the tumour cells was only slightly affected by conditioned media. CONCLUSION: Homotypic collisions between migrating prostatic cancer cells can strongly stimulate their motility. The effect of increased contact is greater on the weakly metastatic cells, such that at high cell density, the difference in the motilities of weak and strong metastatic cells is greatly reduced. PMID- 11890254 TI - Capsaicin effectively prevents apoptosis in the contralateral testis after ipsilateral testicular torsion. AB - OBJECTIVE: To evaluate the effect of afferent nerve blockage by administration of capsaicin on apoptotic changes in the contralateral testis in rats undergoing ipsilateral testicular torsion. MATERIALS AND METHODS: Twenty male albino rats were randomly divided into four groups. In groups 1 and 2, rats underwent a sham operation and testicular torsion, respectively, after the intraperitoneal administration of 0.9% NaCl. Similarly, in groups 3 and 4 the rats underwent a sham operation and testicular torsion, respectively, after an intraperitoneal capsaicin injection. The testes were untwisted 24 h later and the contralateral testes harvested. Apoptosis was assessed in paraffin-embedded sections stained for nuclear DNA fragmentation. Fifteen cells were counted in each seminiferous tubule and the apoptotic cells recorded. A score was calculated for each group and the results compared using the Kruskal-Wallis analysis of variance and Mann Whitney U-tests, with P<0.05 considered to be significant. RESULTS: The mean apoptotic score of group 2 was significantly higher than that of the other groups. There was no difference between the apoptotic scores of groups 1 and 3, 1 and 4, and 3 and 4. CONCLUSION: Capsaicin effectively prevented apoptosis in the contralateral testes of rats that had undergone testicular torsion. PMID- 11890255 TI - Serum chromogranin-A in advanced prostate cancer. AB - OBJECTIVE: To determine the value of serum chromogranin A (CgA), a marker of neuroendocrine differentiation, for monitoring prostate cancer: CgA levels were related to three other tumour markers, i.e. total prostate-specific antigen (tPSA), prostatic acid phosphatase (PAP), neurone-specific enolase (NSE), and to testosterone, to assess the importance of hormone withdrawal. PATIENTS AND METHODS: Serum samples (218) were obtained from 118 patients with prostate cancer, including 111 patients with advanced prostate cancer: 103 presented to our centre for systemic radionuclide therapy of painful skeletal metastases. CgA concentrations were measured using a new immunoradiometric assay (IRMA: Cis Bio International, Gif sur Yvette, France) and a threshold of 70 ng/mL was determined after receiver operating characteristic curve analysis. Testosterone was also measured with an IRMA assay; tPSA, PAP and NSE were assayed using the time resolved amplified cryptate emission. RESULTS: Serum marker levels were high in 64% of the patients for CgA, 24% for NSE, 89% for tPSA and 81% for PAP. Patients resistant to endocrine treatments and with elevated tPSA (i.e. hormone independent) showed increased CgA and NSE in 62% and 29%, respectively. Patients with hormone-dependent prostate cancer (i.e. with a normal tPSA level) had elevated CgA in 59% but no abnormal NSE. All patients of the latter group except one showed clinical progression of their disease. However, the mean (SD) concentrations of CgA in hormone-independent (146) or hormone-dependent (22) patients, at 185.3 (449.1) and 160.9 (293.9) ng/mL, respectively, were not statistically different (P=0.8, Mann-Whitney U-test). For 30 patients, blood samples were drawn and markers measured before and after systemic radionuclide therapy. There was a significant increase in the CgA and tPSA concentrations after treatment (P=0.0146 and 0.0025, respectively). CONCLUSIONS: In association with tPSA, serum CgA appears to be a promising marker for monitoring patients with prostate cancer. PMID- 11890256 TI - Superior transperitoneal dissection for inserting artificial sphincter bladder neck cuffs. PMID- 11890257 TI - Mitrofanoff procedure with Meckel's diverticulum. PMID- 11890258 TI - Nilutamide-induced neutropenia. PMID- 11890259 TI - Ethanol-glycine irrigating fluid for transurethral resection of the prostate in practice. PMID- 11890260 TI - Risk factors for adult renal cell carcinoma: a systematic review and implications for prevention. PMID- 11890261 TI - Urological cancers: do early detection strategies exist? PMID- 11890262 TI - Genetics of the glaucomas. PMID- 11890263 TI - The anatomy and pathophysiology of the optic nerve head in glaucoma. PMID- 11890264 TI - Models of neural injury. AB - A variety of methods can be used experimentally to model neural injury, and thus study the process that represents the major pathophysiology underlying glaucomatous optic neuropathy. These methods vary in difficulty, relevance to human glaucoma, and time course. There is no perfect model, partly because our understanding of the glaucomas is still incomplete. Nonetheless, research into the mechanisms underlying the optic neuropathy of glaucoma (as well as possible new treatments) is advancing rapidly because of the availability of a wide variety of models of neural injury. PMID- 11890265 TI - World health problem of glaucoma. PMID- 11890266 TI - Serum autoantibodies in patients with glaucoma. PMID- 11890267 TI - Screening paradigms in glaucoma. PMID- 11890268 TI - Glaucoma and the brain. PMID- 11890269 TI - Pigmentary glaucoma. PMID- 11890270 TI - Perspective on exfoliation syndrome. PMID- 11890271 TI - Axenfeld-Rieger and iridocorneal endothelial syndromes: two spectra of disease with striking similarities and differences. PMID- 11890272 TI - Diurnal measurement of intraocular pressure. PMID- 11890273 TI - Outflow of aqueous humor. PMID- 11890274 TI - Measurement of uveoscleral outflow in humans. PMID- 11890275 TI - Functional assessment of glaucoma. AB - Several studies have correlated the location of visual field defect on some of these newer tests to location of optic nerve damage with good results. In addition, there is an important and direct relationship between the psychophysical measures of visual function and location of damage. When a given individual had vision loss on more than one test, the same area of the visual field is affected. In addition, as soon as a repeatable defect is identified on perimetry, progression of the defect on later fields occurs within or adjacent to the initially identified area. These findings have significant implications for the care and follow-up of patients with glaucoma. Using these more sensitive tests has improved greatly our detection of early glaucomatous damage. Follow-up testing that concentrates the evaluation to areas already damaged should improve greatly our ability to identify true change from the significant physiologic variability present in glaucoma. PMID- 11890276 TI - Open-angle glaucoma in Japan. PMID- 11890277 TI - Ultrasound biomicroscopy of the anterior segment. PMID- 11890278 TI - Retinal nerve fiber layer evaluation in glaucoma. PMID- 11890279 TI - Assessment of the optic disc in glaucoma. PMID- 11890280 TI - Measurement of ocular blood flow. AB - Many of these techniques have been adapted to investigate hemodynamic alterations, not only in glaucoma, but in diverse disorders such as diabetic retinopathy, macular degeneration, retinal dystrophies, and nonglaucomatous optic neuropathies. However, each of these techniques has limitations and there is not a single methods that provides comprehensive measurements for all the ocular tissues. Our enhanced understanding of ocular vascular anatomy has helped to direct our hemodynamic investigations of the various vascular beds within the eye. However, many of the techniques have lacked validation (reproducibility and accuracy) prior to clinical implementation. Understanding the limitations and the theoretical assumptions of each of these techniques will allow more prudent application in the clinical arena in the future. PMID- 11890281 TI - The influence of corneal thickness on the diagnosis and management of glaucoma. PMID- 11890282 TI - Target pressure. PMID- 11890283 TI - Primary angle-closure glaucoma in Asia. PMID- 11890284 TI - Advances in glaucoma medication during the 1990s and their effects. PMID- 11890285 TI - Maximal glaucoma therapy. PMID- 11890286 TI - Lowering intraocular pressure to minimize glaucoma damage. PMID- 11890287 TI - Strategies for neuroprotection. AB - While glaucoma may be a better candidate for the implementation of neuroprotective strategies than more acute CNS diseases, the failure of clinical neuroprotective trials in stroke should prompt both clinical and basic researchers studying glaucoma to develop better methods to test these agents in animal models, as well as improve methods to quantify glaucomatous damage in clinical studies. The inadequacy of visual fields to quantify glaucomatous progression may hamper current and future clinical trials evaluating neuroprotective agents, and thus may fail to identify potentially beneficial agents and delay the implementation of these strategies. PMID- 11890288 TI - Advances in glaucoma surgery: evolution of antimetabolite adjunctive therapy. PMID- 11890289 TI - Advances with aqueous shunts--1990 to 2001. PMID- 11890290 TI - Nonpenetrating glaucoma surgery. PMID- 11890291 TI - The TIGR/MYOC gene and glaucoma: opportunities for new understandings. PMID- 11890292 TI - Perforating pilomatricoma: transepithelial elimination or not. AB - We present a 56-year-old woman with a perforating pilomatricoma in the left eyebrow region. Histologically, the tumor consisted mainly of basophilic cells and shadow cells, and the tumor components were being eliminated through an ulcer with damage to the epithelial structures. In past reports of perforating pilomatricoma, this elimination pattern has often been described as transepithelial elimination. In many patients with perforating pilomatricoma, elimination is accompanied by ulceration and epithelial damage. Mehregan recently stated that elimination accompanied by epidermal necrosis and superficial ulceration constituted one form of transepithelial elimination. Epidermal necrosis and ulceration generally constitute severe damage. However, when Mehregan first proposed the concept of transepithelial elimination, it was defined as a phenomenon with relatively little or no damage to the epithelial structures, differentiating it from other types of elimination. This original definition makes transepithelial elimination a unique and interesting phenomenon, and its most important feature is that there is relatively little or no damage to the epithelial structures. Therefore, the terms "epidermal necrosis" and "ulceration" should not be used in association with transepithelial elimination. Hence, in patients with perforating pilomatricoma, the elimination of tumor components from ulcers with damage to the epithelial structures, as seen in the present case, should not be described as transepithelial elimination. PMID- 11890293 TI - One confirmed and six suspected cases of cutaneous larva migrans caused by overseas infection with dog hookworm larvae. AB - Cutaneous larva migrans (CLM) is characterized as creeping eruption/serpiginous erythema and/or mobile erythematous induration on the skin. In Japan, Gnathostoma spp. are the most well known pathogens causing CLM, especially the creeping eruption type. Recently, Spirurina type X larvae have been added to the list of causative agents for creeping eruption in Japan. Here we report one confirmed and 6 suspected cases of creeping eruption caused by infection with dog hookworm larvae. The patients were assumed to have been infected overseas. Dog hookworms such as Ancylostoma caninum and A. brasiliense should be considered as possible causative agents for creeping eruption, especially when the patients have a history of travelling overseas. PMID- 11890294 TI - Localized linear scleroderma with cutaneous calcinosis. AB - A 38-year-old woman developed sclerotic and atrophic changes of the left femur in the winter of 1976. In 1980, she was referred to our dermatology clinic and was diagnosed with localized linear scleroderma from the results of the physical examinations and the histological findings. Although several local and systemic treatments were employed over the following 10 years, the sclerotic lesion did not show any remarkable improvement. In 1991, several hard and white papules appeared in the lesion, and a biopsy specimen of these white papules revealed calcinosis. PMID- 11890295 TI - Drug-induced erythroderma with high fever. PMID- 11890296 TI - A case of atypical melanosis of the foot. PMID- 11890297 TI - Clinicopathologic analysis of 66 cases of erythema annulare centrifugum. AB - Erythema annulare centrifugum (EAC) denotes a group of eruptions characterized by slowly migrating annular or configurate erythematous lesions. It can be classified histopathologically as the deep or superficial type. Although there are many case reports of EAC associated with various underlying conditions, no recent clinicopathologic studies exist. The purpose of this study was aimed at analyzing the clinical and histopathologic features of EAC. Sixty-six patients who have been diagnosed as EAC by clinical and histopathological examination were collected. The medical records and skin biopsy specimens of these patients were reviewed retrospectively. There were 24 male patients and 42 female ones. The mean age was 39.7 years, and the mean durtion of the disease was 2.8 years. The lower extremities, particularly the thigh, were the most frequently involved locations. The most common clinical manifestation was large (>1 cm), scaly, erythematous, indurated plaques. Forty-eight patients (72%) had combined diseases including cutaneous fungal infection (48%), such as tinea pedis, other skin diseases (18%), internal malignancies (13%), and other systemic diseases (21%). By histopathologic examination, 33 of 42 patients (78%) were identified as superficial type, and 9 patients (22%) were deep type. Therapeutic trials with systemic or topical corticosteroid and oral antihistamine did not affect the chronic and recurrent course of these patients. EAC is a chronic and recurrent disease despite treatment. EAC is thought to be highly associated with internal disease as well as with superificial fungal infection. However, it was difficult to prove a causal association. The recognition and exact diagnosis of EAC is important, because it may be a quite stressful condition and lead to unnecessary over-treatments. PMID- 11890298 TI - Decreased total numbers of peripheral blood lymphocytes with elevated percentages of CD4+CD45RO+ and CD4+CD25+ of T-helper cells in non-segmental vitiligo. AB - Vitiligo is a disorder involving progressive skin depigmentation caused by host mediated destruction of melanocytes. Its pathogenesis is known to correlate with elevated levels of activated skin-infiltrating T lymphocytes and is presumed to be autoimmune in nature. In the present study, we characterize the immunophenotype of peripheral blood T cells from vitiligo patients, with the objective of developing an investigative and diagnostic tool for the disease, using analysis of peripheral blood. Subjects for this investigation included 32 patients diagnosed with non-segmental vitiligo and 28 age-and gender-matched, normal, healthy control participants. Whole venous blood taken from each subject was analyzed using 2-color flow cytometry for immunologically-relevant lymphocyte subsets. When compared with healthy control subjects, peripheral blood from individuals with vitiligo was found to have lower total numbers of lymphocytes (p<0.039). Vitiligo patients also had elevated percentages of memory (CD4+CD45RO+) T cells; (p<0.05), but NK-T cells (CD3+CD16+CD56+) and naive T cells (CD4+CD45RA+) were present at lower total numbers and percentages than in healthy controls (p<0.01 and 0.05 respectively). Blood from severely afflicted subjects exhibited elevated CD3+HLADR+ and CD4+CD45RO+ as well as lower percentages of NK-T cells (p<0.05) when compared with mild cases. In conclusion, disease-associated, peripheral blood lymphocyte immunophenotypic profiles of vitiligo patients are consistent with the hypothesis of T cell activation as a major feature of the disorder. These include elevated memory and reduced naive T cell percentages and increased expression of the activation-associated surface antigen CD25. These changes presumably reflect increased antigen-mediated activation. Moreover, because a corollary effect is increased activation-induced cell death (AICD), lower overall lymphocyte counts observed in vitiligo-afflicted subjects is also expected. PMID- 11890299 TI - Rising incidence of genital herpes over two decades in a sexually transmitted disease clinic in north India. AB - Genital herpes, which was considered to be a minor sexually transmitted disease (STD) in the past in developing countries, is rapidly increasing; in contrast, bacterial STDs are declining. This changing trend of various STDs prompted us to analyze our data retrospectively to see whether a similar change is occurring in this part of India as well. The records of sexually transmitted diseases (STDs) clinic attendees from January 1977 to December 2000 were analyzed. The demographic profile of patients with genital herpes was also considered for analysis. STDs were diagnosed clinically and by appropriate laboratory tests wherever applicable. VDRL test was done in all patients, and HIV antibody detection was performed from 1987 onwards. The incidences of chancroid, donovanosis, and gonorrhea were 12.2%, 6.3%, and 16.9%, respectively, from January 1977 to December 1985. The figures for the same decreased to 2.5%, 0.9%, and 2.3%, respectively, from January 1993 to December 2000. The decreasing incidence of the above bacterial STDs is statistically significant (p<0.001). However, there was an approximately two-fold increase in the incidence of genital herpes in recent years (20.5%) in comparison to the figures from the late 70s (11.4%). Molluscum contagiosum also showed an upward trend (1% in 1977-85 vs. 9.8% in 1993-2000). Condylomata accuminata remained almost unchanged (21.4% in 1977-85 vs. 20% in 1993-2000). To conclude, a significant increase in the number of viral STDs and a decline in the bacterial diseases were observed in recent years in comparison to the figures from the late 70s. This may be due to awareness of HIV, success of control programs, syndromic management of STDs, and adoption of safer sexual practices, which prevent bacterial STDs more efficiently than viral ones. PMID- 11890300 TI - A case of eruptive collagenoma localized on the neck and shoulders. AB - A 78-year-old woman, who had first noticed asymptomatic eruptions on her neck and shoulders eight years earlier, presented with papules and nodules 2 to 20 mm in diameter that had a normal to white hue and were flatly elevated. These lesions were scattered and multiple, some forming confluent plaques. Histopathologically, the epidermis was slightly atrophied, and collagen fibers in the dermis were coarse and proliferated. In addition, the number of elastic fibers was slightly decreased. No complications were evident. Based on these findings, the patient was given a diagnosis of mild eruptive collagenoma, a type of connective tissue nevus according to the classification of Uitto. This case is unique in that onset was at an advanced age and that distribution was localized on the neck and shoulders. PMID- 11890301 TI - Pyoderma gangrenosum associated with paroxysmal nocturnal hemoglobulinuria and monoclonal gammopathy. AB - Pyoderma gangrenosum developed in a man with a five-year history of paroxysmal nocturnal hemoglobinuria and monoclonal gammopathy. He had multiple walnut sized ulcers on his back and extremities, plasma IgM-k type M-protein and low erythrocytic CD55 expression. This is an extremely rare association. However, clonal expansion of plasma cells and chimeric expression of hematopoietic cell glycosylphosphatidylinositol (GPI)-anchored proteins may represent somatic mutations of hematopoietic stem cells in PG as well as PNH. PNH is based on abnormalities in the GPI-anchor formation on various hematopoietic and non hematopoietic cells. Since the GPI-anchored proteins have pleiotropic functions in complement mediated cell lysis, leukocyte motility, and coagulation systems, the present case may indicate the possible involvement of a GPI-anchored protein abnormality in the pathogenesis of PG. PMID- 11890302 TI - Sweet's syndrome evolved from recurrent erythema nodosum in a patient with myelodysplastic syndrome. AB - A 63-year-old man had painful nodules on his lower legs. Microscopic examination showed septal and lobular panniculitis composed of lympho-histiocytic infiltrates. Based on the clinical and histopathological findings, the diagnosis of erythema nodosum (EN) was made. Nonsteroidal anti-inflammatory drugs were temporarily effective, but the eruptions had repeated to the present, and 16 months later, myelodysplastic syndrome (MDS) was diagnosed. Then, 6 months later, he developed a high fever and edematous fresh red-colored nodules on his neck, arm and upper trunk. Histopathologically, a diffuse, dense, dermal infiltrate of neutrophils was seen, and Sweet's syndrome (SS) was diagnosed. SS is known to develop in patients with MDS, and EN is one of the dermatoses that occur in conjunction with hematoproliferative disorders. Furthermore, SS evolving from recurrent EN and the simultaneous occurrence of SS and EN have been reported in some patients. In our case, we suggest that some mediators such as cytokines associated with MDS might have first induced EN, and then, as the MDS developed, they were replaced by others that caused SS. PMID- 11890303 TI - A case of chronic perianal pyoderma treated with the recycled skin graft method. AB - We present here a case of chronic perianal pyoderma (CPP), who was treated with a novel skin graft. The patient was a 28-year-old male who had suffered from painful abscesses and nodules on his buttocks for over 10 years. Although the abscesses and nodules were initially restricted to one buttock, they gradually spread over both buttocks. He visited our hospital in July of 2000. He was clinically diagnosed as CPP and treated with oral doses of antibiotics. When admitted to our hospital in September of 2000, an anal fistula was also found. The operations for CPP and anal fistula were simultaneously performed. After the anal fistula treatment, the lesions of CPP were totally resected. In this procedure, we removed the epidermis and upper dermis from the excised CPP lesions and grafted them on the defects of the excised lesion. There has been no recurrence of the CPP. This operation procedure, which we call the "recycled skin graft method" is less invasive, because a donor site for the skin graft is unnecessary. We considered it to be as one of the effective treatments for CPP. PMID- 11890304 TI - Model-checking techniques based on cumulative residuals. AB - Residuals have long been used for graphical and numerical examinations of the adequacy of regression models. Conventional residual analysis based on the plots of raw residuals or their smoothed curves is highly subjective, whereas most numerical goodness-of-fit tests provide little information about the nature of model misspecification. In this paper, we develop objective and informative model checking techniques by taking the cumulative sums of residuals over certain coordinates (e.g., covariates or fitted values) or by considering some related aggregates of residuals, such as moving sums and moving averages. For a variety of statistical models and data structures, including generalized linear models with independent or dependent observations, the distributions of these stochastic processes tinder the assumed model can be approximated by the distributions of certain zero-mean Gaussian processes whose realizations can be easily generated by computer simulation. Each observed process can then be compared, both graphically and numerically, with a number of realizations from the Gaussian process. Such comparisons enable one to assess objectively whether a trend seen in a residual plot reflects model misspecification or natural variation. The proposed techniques are particularly useful in checking the functional form of a covariate and the link function. Illustrations with several medical studies are provided. PMID- 11890305 TI - Combining complete multivariate outcomes with incomplete covariate information: a latent class approach. AB - This work was motivated by the need to combine outcome information from a reference population with risk factor information from a screened subpopulation in a setting where the analytic goal was to study the association between risk factors and multiple binary outcomes. To achieve such an analytic goal, this article proposes a two-stage latent class procedure that first summarizes the commonalities among outcomes using a reference population sample, then analyzes the association between outcomes and risk factors. It develops a pseudo-maximum likelihood approach to estimating model parameters. The performance of the proposed method is evaluated in a simulation study and in an illustrative analysis of data from the Women's Health and Aging Study, a recent investigation of the causes and course of disability in older women. Combining information in the proposed way is found to improve both accuracy and precision in summarizing multiple categorical outcomes, which effectively diminishes ambiguity and bias in making risk factor inferences. PMID- 11890306 TI - Functional mixed effects models. AB - In this article, a new class of functional models in which smoothing splines are used to model fixed effects as well as random effects is introduced. The linear mixed effects models are extended to nonparametric mixed effects models by introducing functional random effects, which are modeled as realizations of zero mean stochastic processes. The fixed functional effects and the random functional effects are modeled in the same functional space, which guarantee the population average and subject-specific curves have the same smoothness property. These models inherit the flexibility of the linear mixed effects models in handling complex designs and correlation structures, can include continuous covariates as well as dummy factors in both the fixed or random design matrices, and include the nested curves models as special cases. Two estimation procedures are proposed. The first estimation procedure exploits the connection between linear mixed effects models and smoothing splines and can be fitted using existing software. The second procedure is a sequential estimation procedure using Kalman filtering. This algorithm avoids inversion of large dimensional matrices and therefore can be applied to large data sets. A generalized maximum likelihood (GML) ratio test is proposed for inference and model selection. An application to comparison of cortisol profiles is used as an illustration. PMID- 11890307 TI - On estimation and prediction for spatial generalized linear mixed models. AB - We use spatial generalized linear mixed models (GLMM) to model non-Gaussian spatial variables that are observed at sampling locations in a continuous area. In many applications, prediction of random effects in a spatial GLMM is of great practical interest. We show that the minimum mean-squared error (MMSE) prediction can be done in a linear fashion in spatial GLMMs analogous to linear kriging. We develop a Monte Carlo version of the EM gradient algorithm for maximum likelihood estimation of model parameters. A by-product of this approach is that it also produces the MMSE estimates for the realized random effects at the sampled sites. This method is illustrated through a simulation study and is also applied to a real data set on plant root diseases to obtain a map of disease severity that can facilitate the practice of precision agriculture. PMID- 11890308 TI - Semiparametric regression modeling with mixtures of Berkson and classical error, with application to fallout from the Nevada test site. AB - We construct Bayesian methods for semiparametric modeling of a monotonic regression function when the predictors are measured with classical error. Berkson error, or a mixture of the two. Such methods require a distribution for the unobserved (latent) predictor, a distribution we also model semiparametrically. Such combinations of semiparametric methods for the dose response as well as the latent variable distribution have not been considered in the measurement error literature for any form of measurement error. In addition, our methods represent a new approach to those problems where the measurement error combines Berkson and classical components. While the methods are general, we develop them around a specific application, namely, the study of thyroid disease in relation to radiation fallout from the Nevada test site. We use this data to illustrate our methods, which suggest a point estimate (posterior mean) of relative risk at high doses nearly double that of previous analyses but that also suggest much greater uncertainty in the relative risk. PMID- 11890309 TI - Mixed effects logistic regression models for multiple longitudinal binary functional limitation responses with informative drop-out and confounding by baseline outcomes. AB - In the context of analyzing multiple functional limitation responses collected longitudinally from the Longitudinal Study of Aging (LSOA), we investigate the heterogeneity of these outcomes with respect to their associations with previous functional status and other risk factors in the presence of informative drop-out and confounding by baseline outcomes. We accommodate the longitudinal nature of the multiple outcomes with a unique extension of the nested random effects logistic model with an autoregressive structure to include drop-out and baseline outcome components with shared random effects. Estimation of fixed effects and variance components is by maximum likelihood with numerical integration. This shared parameter selection model assumes that drop-out is conditionally independent of the multiple functional limitation outcomes given the underlying random effect representing an individual's trajectory of functional status across time. Whereas it is not possible to fully assess the adequacy of this assumption, we assess the robustness of this approach by varying the assumptions underlying the proposed model such as the random effects structure, the drop-out component, and omission of baseline functional outcomes as dependent variables in the model. Heterogeneity among the associations between each functional limitation outcome and a set of risk factors for functional limitation, such as previous functional limitation and physical activity, exists for the LSOA data of interest. Less heterogeneity is observed among the estimates of time-level random effects variance components that are allowed to vary across functional outcomes and time. We also note that. under an autoregressive structure, bias results from omitting the baseline outcome component linked to the follow-up outcome component by subject-level random effects. PMID- 11890311 TI - Generalized character process models: estimating the genetic basis of traits that cannot be observed and that change with age or environmental conditions. AB - The genetic analysis of characters that change as a function of some independent and continuous variable has received increasing attention in the biological and statistical literature. Previous work in this area has focused on the analysis of normally distributed characters that are directly observed. We propose a framework for the development and specification of models for a quantitative genetic analysis of function-valued characters that are not directly observed, such as genetic variation in age-specific mortality rates or complex threshold characters. We employ a hybrid Markov chain Monte Carlo algorithm involving a Monte Carlo EM algorithm coupled with a Markov chain approximation to the likelihood, which is quite robust and provides accurate estimates of the parameters in our models. The methods are investigated using simulated data and are applied to a large data set measuring mortality rates in the fruit fly, Drosophila melanogaster. PMID- 11890310 TI - Characterizing the relationship between HIV-1 genotype and phenotype: prediction based classification. AB - This paper establishes a framework for understanding the complex relationships between HIV-1 genotypic markers of resistance to antiretroviral drugs and clinical measures of disease progression. A new classification scheme based on the probabilities of how new patients will respond to antiretroviral therapy given the available data is proposed as a method for distinguishing among groups of viral sequences. This approach draws from existing cluster analysis, discriminant analysis, and recursive partitioning techniques and requires a model relating genotypic characteristics to phenotypic response. A data set of 2,746 sequences and the corresponding Indinavir 50% inhibitory concentrations are described and used for illustrative purposes. PMID- 11890312 TI - Two-stage designs for gene-disease association studies. AB - The goal of this article is to describe a two-stage design that maximizes the power to detect gene-disease associations when the principal design constraint is the total cost, represented by the total number of gene evaluations rather than the total number of individuals. In the first stage, all genes of interest are evaluated on a subset of individuals. The most promising genes are then evaluated on additional subjects in the second stage. This will eliminate wastage of resources on genes unlikely to be associated with disease based on the results of the first stage. We consider the case where the genes are correlated and the case where the genes are independent. Using simulation results, it is shown that, as a general guideline when the genes are independent or when the correlation is small, utilizing 75% of the resources in stage 1 to screen all the markers and evaluating the most promising 10% of the markers with the remaining resources provides near-optimal power for a broad range of parametric configurations. This translates to screening all the markers on approximately one quarter of the required sample size in stage 1. PMID- 11890313 TI - Dose finding using the biased coin up-and-down design and isotonic regression. AB - We are interested in finding a dose that has a prespecified toxicity rate in the target population. In this article, we investigate five estimators of the target dose to be used with the up-and-down biased coin design (BCD) introduced by Durham and Flournoy (1994, Statistical Decision Theory and Related Topics). These estimators are derived using maximum likelihood, weighted least squares, sample averages, and isotonic regression. A linearly interpolated isotonic regression estimate is shown to be simple to derive and to perform as well as or better than the other target dose estimators in terms of mean square error and average number of subjects needed for convergence in most scenarios studied. PMID- 11890314 TI - Effect of list errors on the estimation of population size. AB - In many situations, it is possible to estimate the size of a closed population if some members of the population are recorded on one or more administrative lists. An important example is estimating the prevalence of a disease, where some members of the disease population may be recorded on lists such as disease registries, hospital admission records, and general practitioner records. An incomplete contingency table is formed by matching the lists and the missing cell count estimated by prediction based on a fitted model, which assumes that the matching is done without error. In practice, matching errors do occur, and in this article, we examine the effect of these errors on the estimation process and show how the standard models may be modified to allow for matching errors. PMID- 11890315 TI - Testing for conditional multiple marginal independence. AB - Survey respondents are often prompted to pick any number of responses from a set of possible responses. Categorical variables that summarize this kind of data are called pick any/c variables. Counts from surveys that contain a pick any/c variable along with a group variable (r levels) and stratification variable (q levels) can be marginally summarized into an r x c x q contingency table. A question that may naturally arise from this setup is to determine if the group and pick any/c variable are marginally independent given the stratification variable. A test for conditional multiple marginal independence (CMMI) can be used to answer this question. Since subjects may pick any number out of c possible responses, the Cochran (1954, Biometrics 10, 417-451) and Mantel and Haenszel (1959, Journal of the National Cancer Institute 22, 719-748) tests cannot be used directly because they assume that units in the contingency table are independent of each other. Therefore, new testing methods are developed. Cochran's test statistic is extended to r x 2 x q tables, and a modified version of this statistic is proposed to test CMMI. Its sampling distribution can be approximated through bootstrapping. Other CMMI testing methods discussed are bootstrap p-value combination methods and Bonferroni adjustments. Simulation findings suggest that the proposed bootstrap procedures and the Bonferroni adjustments consistently hold the correct size and provide power against various alternatives. PMID- 11890316 TI - Interval estimation for a difference between intraclass kappa statistics. AB - Model-based inference procedures for the kappa statistic have developed rapidly over the last decade. However, no method has yet been developed for constructing a confidence interval about a difference between independent kappa statistics that is valid in samples of small to moderate size. In this article, we propose and evaluate two such methods based on an idea proposed by Newcombe (1998, Statistics in Medicine, 17, 873-890) for constructing a confidence interval for a difference between independent proportions. The methods are shown to provide very satisfactory results in sample sizes as small as 25 subjects per group. Sample size requirements that achieve a prespecified expected width for a confidence interval about a difference of kappa statistic are also presented. PMID- 11890317 TI - Principal stratification in causal inference. AB - Many scientific problems require that treatment comparisons be adjusted for posttreatment variables, but the estimands underlying standard methods are not causal effects. To address this deficiency, we propose a general framework for comparing treatments adjusting for posttreatment variables that yields principal effects based on principal stratification. Principal stratification with respect to a posttreatment variable is a cross-classification of subjects defined by the joint potential values of that posttreatment variable tinder each of the treatments being compared. Principal effects are causal effects within a principal stratum. The key property of principal strata is that they are not affected by treatment assignment and therefore can be used just as any pretreatment covariate. such as age category. As a result, the central property of our principal effects is that they are always causal effects and do not suffer from the complications of standard posttreatment-adjusted estimands. We discuss briefly that such principal causal effects are the link between three recent applications with adjustment for posttreatment variables: (i) treatment noncompliance, (ii) missing outcomes (dropout) following treatment noncompliance. and (iii) censoring by death. We then attack the problem of surrogate or biomarker endpoints, where we show, using principal causal effects, that all current definitions of surrogacy, even when perfectly true, do not generally have the desired interpretation as causal effects of treatment on outcome. We go on to forrmulate estimands based on principal stratification and principal causal effects and show their superiority. PMID- 11890318 TI - Levene tests of homogeneity of variance for general block and treatment designs. AB - This article develops a weighted least squares version of Levene's test of homogeneity of variance for a general design, available both for univariate and multivariate situations. When the design is balanced, the univariate and two common multivariate test statistics turn out to be proportional to the corresponding ordinary least squares test statistics obtained from an analysis of variance of the absolute values of the standardized mean-based residuals from the original analysis of the data. The constant of proportionality is simply a design dependent multiplier (which does not necessarily tend to unity). Explicit results are presented for randomized block and Latin square designs and are illustrated for factorial treatment designs and split-plot experiments. The distribution of the univariate test statistic is close to a standard F-distribution, although it can be slightly underdispersed. For a complex design, the test assesses homogeneity of variance across blocks, treatments, or treatment factors and offers an objective interpretation of residual plots. PMID- 11890319 TI - Dynamic conditionally linear mixed models for longitudinal data. AB - We develop a new class of models, dynamic conditionally linear mixed models, for longitudinal data by decomposing the within-subject covariance matrix using a special Cholesky decomposition. Here 'dynamic' means using past responses as covariates and 'conditional linearity' means that parameters entering the model linearly may be random, but nonlinear parameters are nonrandom. This setup offers several advantages and is surprisingly similar to models obtained from the first order linearization method applied to nonlinear mixed models. First, it allows for flexible and computationally tractable models that include a wide array of covariance structures; these structures may depend on covariates and hence may differ across subjects. This class of models includes, e.g., all standard linear mixed models, antedependence models, and Vonesh-Carter models. Second, it guarantees the fitted marginal covariance matrix of the data is positive definite. We develop methods for Bayesian inference and motivate the usefulness of these models using a series of longitudinal depression studies for which the features of these new models are well suited. PMID- 11890320 TI - The use of frailty hazard models for unrecognized heterogeneity that interacts with treatment: considerations of efficiency and power. AB - Increasingly, genetic studies of tumors of the same histologic diagnosis are elucidating subtypes that are distinct with respect to clinical endpoints such as response to treatment and survival. This raises concerns about the efficiency of using the simple log-rank test for analysis of treatment effect on survival in studies of possibly heterogeneous tumors. Furthermore, such studies, designed under the assumption of homogeneity, may be severely underpowered. We derive analytic approximations for the asymptotic relative efficiency of the simple log rank test relative to the optimally weighted log-rank test and for the power of the simple log-rank test when applied to subjects with unobserved heterogeneity, as reflected in a continuous frailty, that may interact with treatment. Numerical studies demonstrate that the simple log-rank test may be quite inefficient if the frailty interacts with treatment. Further, there may be a substantial loss of power in the presence of the frailty with or without an interaction with treatment. PMID- 11890321 TI - On the use of nonparametric curves in phase I trials with low toxicity tolerance. AB - Gasparini and Eisele (2000, Biometrics 56, 609-615) propose a design for phase I clinical trials during which dose allocation is governed by a Bayesian nonparametric estimate of the dose-response curve. The authors also suggest an elicitation algorithm to establish vague priors. However, in situations where a low percentile is targeted, priors thus obtained can lead to undesirable rigidity given certain trial outcomes that can occur with a nonnegligible probability. Interestingly, improvement can be achieved by prescribing slightly more informative priors. Some guidelines for prior elicitation are established using a connection between this curve-free method and the continual reassessment method. PMID- 11890322 TI - The DNA database search controversy. AB - A recent article in Biometrics (Stockmarr, 1999, 55, 671-677) has generated correspondence (56, 1274-1277; 57, 976-980) reigniting a controversy started by a 1996 report on DNA profile evidence issued by the U.S. National Research Council (NRC). The issue concerns the evidential weight of a DNA profile match when the match results from a search through a profile database. The views of both Stockmarr and the NRC report conflict with those of many statisticians working in the area, and the differing viewpoints lead to dramatically different assessments of evidence. I outline reasons why Stockmarr and the NRC report are wrong. I also briefly discuss possible reasons why forensic applications tend to be problematic for statisticians. PMID- 11890323 TI - Curve-free and model-based continual reassessment method designs. AB - Gasparini and Eisele (2000, Biometrics 56, 609 615) present a development of the continual reassessment method of O'Quigley, Pepe, and Fisher (1990, Biometrics 46, 33-48). They call their development a curve-free method for Phase I clinical trials. However, unless we are dealing with informative prior information, then the curve-free method coincides with the usual model-based continual reassessment method. Both methods are subject to arbitrary specification parameters, and we provide some discussion on this. Whatever choices are made for one method, there exists equivalent choices for the other method, where " equivalent" means that the operating characteristics (sequential dose allocation and final recommendation) are the same. The insightful development of Gasparini and Eisele provides clarification on some of the basic ideas behind the continual reassessment method, particularly when viewed from a Bayesian perspective. But their development does not lead to a new class of designs and the comparative results in their article, indicating some preference for curve-free designs over model-based designs, are simply reflecting a more fortunate choice of arbitrary specification parameters. Other choices could equally well have inversed their conclusion. A correct conclusion should be one of operational equivalence. The story is different for the case of informative priors, a situation that is inherently much more difficult. We discuss this. We also mention the important idea of two-stage designs (Moller, 1995, Statistics in Medicine 14, 911-922; O'Quigley and Shen, 1996, Biometrics 52, 163-174), arguing, via a simple comparison with the results of Gasparini and Eisele (2000), that there is room for notable gains here. Two-stage designs also have an advantage of avoiding the issue of prior specification altogether. PMID- 11890324 TI - The Baumgartner-Weiss-Schindler test in the presence of ties. PMID- 11890325 TI - Modeling and optimization in early detection programs with a single exam. AB - The choice of timing of screening examinations is an important element in determining the efficacy of strategies for the early detection of occult disease. In this article, we describe a flexible decision-making framework for the design of early detection programs, and we investigate the choice of timing when each individual in the screening program is examined only once. We focus on the theoretical relation between the optimal examination time and the distributions of sojourn times in health-related states. Specifically, we derive closed-form solutions of the optimal age using two specifications of utility functions, discuss the effects of natural history and utility specifications on the optimal solution, and present an application to early detection of colorectal cancer by once-only sigmoidoscopy or colonoscopy. PMID- 11890326 TI - Estimation of survival distributions of treatment policies in two-stage randomization designs in clinical trials. AB - Some clinical trials follow a design where patients are randomized to a primary therapy at entry followed by another randomization to maintenance therapy contingent upon disease remission. Ideally, analysis would allow different treatment policies, i.e., combinations of primary and maintenance therapy if specified up-front, to be compared. Standard practice is to conduct separate analyses for the primary and follow-up treatments, which does not address this issue directly. We propose consistent estimators for the survival distribution and mean restricted survival time for each treatment policy in such two-stage studies and derive large-sample properties. The methods are demonstrated on a leukemia clinical trial data set and through simulation. PMID- 11890327 TI - Testing for dependence between failure time and visit compliance with interval censored data. AB - Interval-censored failure-time data arise when subjects miss prescheduled visits at which the failure is to be assessed. The resulting intervals in which the failure is known to have occurred are overlapping. Most approaches to the analysis of these data assume that the visit-compliance process is ignorable with respect to likelihood analysis of the failure-time distribution. While this assumption offers considerable simplification, it is not always plausible. Here we test for dependence between the failure- and visit-compliance processes, applicable to studies in which data collection continues after the occurrence of the failure. We do not make any of the assumptions made by previous authors about the joint distribution of the visit-compliance process, a covariate process, and the failure time. Instead, we consider conditional models of the true failure history given the current visit compliance at each visit time, allowing for correlation across visit times. Because failure status is not known at some visit times due to missed visits, only models of the observed failure history given current visit compliance are estimable. We describe how the parameters from these models can be used to test for a negative association and how bounds on unestimable parameters provided by the observed data are needed additionally to infer a positive association. We illustrate the method with data from an AIDS study and we investigate the power of the test through a simulation study. PMID- 11890328 TI - Estimation in the cox proportional hazards model with left-truncated and interval censored data. AB - We show that the nonparametric maximum likelihood estimate (NPMLE) of the regression coefficient from the joint likelihood (of the regression coefficient and the baseline survival) works well for the Cox proportional hazards model with left-truncated and interval-censored data, but the NPMLE may underestimate the baseline survival. Two alternatives are also considered: first, the marginal likelihood approach by extending Satten (1996, Biometrika 83, 355-370) to truncated data, where the baseline distribution is eliminated as a nuisance parameter; and second, the monotone maximum likelihood estimate that maximizes the joint likelihood by assuming that the baseline distribution has a nondecreasing hazard function, which was originally proposed to overcome the underestimation of the survival from the NPMLE for left-truncated data without covariates (Tsai, 1988, Biometrika 75, 319-324). The bootstrap is proposed to draw inference. Simulations were conducted to assess their performance. The methods are applied to the Massachusetts Health Care Panel Study data set to compare the probabilities of losing functional independence for male and female seniors. PMID- 11890329 TI - A proportional hazards model for incidence and induced remission of disease. AB - To assess the protective effects of a time-varying covariate, we develop a stochastic model based on tumor biology. The model assumes that individuals have a Poisson-distributed pool of initiated clones, which progress through predetectable, detectable mortal and detectable immortal stages. Time-independent covariates are incorporated through a log-linear model for the expected number of clones, resulting in a proportional hazards model for disease onset. By allowing time-dependent covariates to induce clone death, with rate dependent on a clone's state, the model is flexible enough to accommodate delayed disease onset and remission or cure of preexisting disease. Inference uses Bayesian methods via Markov chain Monte Carlo. Theoretical properties are derived, and the approach is illustrated through analysis of the effects of childbirth on uterine leiomyoma (fibroids). PMID- 11890330 TI - Bayesian models for multivariate current status data with informative censoring. AB - Multivariate current status data, consist of indicators of whether each of several events occur by the time of a single examination. Our interest focuses on inferences about the joint distribution of the event times. Conventional methods for analysis of multiple event-time data cannot be used because all of the event times are censored and censoring may be informative. Within a given subject, we account for correlated event times through a subject-specific latent variable, conditional upon which the various events are assumed to occur independently. We also assume that each event contributes independently to the hazard of censoring. Nonparametric step functions are used to characterize the baseline distributions of the different event times and of the examination times. Covariate and subject specific effects are incorporated through generalized linear models. A Markov chain Monte Carlo algorithm is described for estimation of the posterior distributions of the unknowns. The methods are illustrated through application to multiple tumor site data from an animal carcinogenicity study. PMID- 11890331 TI - Monitoring the rates of composite events with censored data in phase II clinical trials. AB - In many phase II clinical trials, interim monitoring is based on the probability of a binary event, response, defined in terms of one or more time-to-event variables within a time period of fixed length. Such outcome-adaptive methods may require repeated interim suspension of accrual in order to follow each patient for the time period required to evaluate response. This may increase trial duration, and eligible patients arriving during such delays either must wait for accrual to reopen or be treated outside the trial. Alternatively, monitoring may be done continuously by ignoring censored data each time the stopping rule is applied, which wastes information. We propose an adaptive Bayesian method that eliminates these problems. At each patient's accrual time, an approximate posterior for the response probability based on all of the event-time data is used to compute an early stopping criterion. Application to a leukemia trial with a composite event shows that the method can reduce trial duration substantially while maintaining the reliability of interim decisions. PMID- 11890332 TI - Frailty models with missing covariates. AB - We present a method for estimating the parameters in random effects models for survival data when covariates are subject to missingness. Our method is more general than the usual frailty model as it accommodates a wide range of distributions for the random effects, which are included as an offset in the linear predictor in a manner analogous to that used in generalized linear mixed models. We propose using a Monte Carlo EM algorithm along with the Gibbs sampler to obtain parameter estimates. This method is useful in reducing the bias that may be incurred using complete-case methods in this setting. The methodology is applied to data from Eastern Cooperative Oncology Group melanoma clinical trials in which observations were believed to be clustered and several tumor characteristics were not always observed. PMID- 11890333 TI - Surgical treatment of schistosomal portal hypertension. AB - Schistosomiasis mansoni is a widespread parasitic disease in the Brazilian territory that affects over 8 million individuals. Hepatosplenic schistosomiasis is a serious clinical presentation of this disease, associated with splenomegaly, liver fibrosis, and portal hypertension, and is responsible for approximately 7% of schistosomotic patients. The surgical treatment of portal hypertension in schistosomotic patients has distinct features when compared with cirrhotic patients, mostly because hepatic function is preserved in schistosomotic liver disease. Therefore, when attempting to reduce the portal pressure, the surgeon must be aware that the surgery might interfere with hepatic perfusion, and consequently with hepatic function. The aim of this study was to report the results achieved with splenectomy, division of the left gastric vein, devascularization of great gastric curvature, and postoperative endoscopic variceal sclerosis, as a surgical option to esophageal varices in hepatosplenic schistosomiasis. A total of 111 patients were studied, and the following is a list of inclusion criteria: age >16 years, history of gastrointestinal (GI) bleeding, presence of esophageal varices on preoperative endoscopy, hematocrit >22% and prothrombin enzymatic activity >50%, negative viral hepatitis on serologic tests (anti-HBV and anti-HCV), and definition, after liver biopsy, of exclusive schistosomotic liver disease. The following list includes exclusion criteria used: presence of liver disease other than schistosomotic, history of alcohol abuse, and preoperative thrombosis of the portal vein. The rebleeding rate was 14.4% during a mean 30-month follow-up period; portal vein thrombosis was 13.2%, and there was a global mortality of 5.4%. Gastric varices were present in 46.9% of the patients; for those patients, a gastrotomy and running suture of the varices achieved an eradication rate of the varices of 75.6%. The degree of periportal fibrosis was also analyzed. Periportal fibrosis staging revealed that patients with class II or III liver fibrosis had a significant increased risk of recurrent GI bleeding when compared with patients with class I liver fibrosis. Despite the elevation on alanine aminotransferase (ALT) and aspartate aminotransferase (AST), most other liver function tests showed no alteration or were corrected after surgery. We conclude that splenectomy, division of the left gastric vein, devascularization of great gastric curvature, and postoperative endoscopic variceal sclerosis showed good results globally and should be considered as therapeutic options in the treatment of hepatosplenic schistosomiasis. PMID- 11890334 TI - Lymph node metastases in gastric cancer: correlation between new and old UICC TNM classification. AB - The 5th edition of TNM classification (1997) grades lymph node involvement in gastric cancer by the number of metastatic lymph nodes. Their prognostic significance as defined by the new (5th edn., 1997) and old (4th edn., 1987) TNM classification was evaluated and survival in pN categories between both versions was compared. It was demonstrated in our analysis that comparison of old and new TNM systems is possible. Categories pN1 and pN2 contain patients selected by different criteria in both versions of TNM classification but with similar survival probabilities. Anatomic location of lymph nodes as described in the 4th edn. and number of involved nodes in the 5th edn. of TNM classification have about the same prognostic value in categories pN1 and pN2. The advantage of the 5th edn. is the identification of a group of patients (pN3, >15 involved lymph nodes) with significantly poor prognosis, which, in our series, includes 15% of R0 resected patients with lymph node metastases. PMID- 11890335 TI - Primary non-Hodgkin lymphoma of the right colon: a retrospective clinical pathological study. AB - The objective of this study was to analyze the results of surgical treatment of primary non-Hodgkin lymphomas of the right colon. Ten patients were operated on with curative intention. Dawson's criteria were used to characterize the colonic lymphoma as a primary lymphomas. In the staging of the tumor, the Ann Arbor classification for gastrointestinal lymphomas modified by Musshoff and Schmidt Vollmer was used. The histological classification was made by using the International Working Formulation Group system. All patients were submitted to radical right colectomy and 6 of them received postoperative chemotherapy. The overall average survival was 39.2 months. Four of the patients are still alive, without active disease, with an average survival of 85.2 months. Six patients died due to relapse in the abdomen, with an average survival of 8.2 months. These results suggest that it is advantageous to patient survival to have them submitted for resection of their lesions at an initial stage of the disease (IE and IIE1). Chemotherapy must be used as a complementary treatment in locally advanced lesions, in an attempt to control the residual microscopic disease. PMID- 11890336 TI - Gut-endocrinomas (carcinoids and related endocrine variants) of the breast: an analysis of 310 reported cases. AB - The purpose of this study was to analyze the present status of gut-endocrinomas (carcinoids and related endocrine variants) of the breast, an extremely rare site for primary growth of such neoplasms, and to provide precise and reliable information concerning these neoplasms on varying clinicopathological aspects for investigators engaged in relevant research fields. A total of 310 cases presented in this analysis consisted of 196 carcinoids, 102 typical and 94 atypical, and 114 related endocrine variants; in the last group, the expression of "breast carcinoma with (neuro-) endocrine differentiation" was often used without referring to the term "carcinoid." A statistical evaluation was performed on most occasions based on a comparison among three groups of typical carcinoids, atypical carcinoids and related endocrine variants, or between the former two series of carcinoids and the third series of endocrine carcinomas. Statistically significant differences between the groups of carcinoids and endocrine carcinomas were recognized in terms of average age, tumor size categories of < or = 20 mm and 21-50 mm, rates of metastases, and positive neuron-specific enolase (NSE) immunohistochemistry. Contrary to our expectations no statistically significant difference between these two groups was evident in terms of overall average tumor size, Grimelius argyrophilia for endocrine nature, or postoperative 5-year survival rates in curative resection cases. It seems important to establish more precise diagnostic criteria for "endocrine carcinomas" from the viewpoint of a certain possibility that some of these neoplasms may belong to the atypical carcinoid group. PMID- 11890337 TI - Vascular complications in the first 100 liver transplantations in Saudi Arabia. AB - Vascular complications are a significant cause of morbidity and mortality following liver transplantation. The purpose of this study is to determine the impact of vascular complications on the patient and graft survival. One hundred liver transplant procedures were performed on 92 patients between January 1994 and June 1998 at King Fahad Hospital (Riyadh, Saudi Arabia). Patients' data were analyzed retrospectively with special emphasis on vascular complications. Vascular complication occurred in 19 procedures (19%), including HAT (n = 8), IVC stenosis (n = 3), PVT (n = 2), HAS (n = 2), rupture HA mycotic aneurysm (n = 1), and celiac artery stenosis (n = 1). Combined vascular complication occurred in 2 patients (HAT and PVT, and HAS and IVC stenosis). The only significant risk factor for the development of vascular thrombosis in our study was pediatric age. The use of low-dose heparin was found to be a protective factor against HAT (P = 0.03). Complications were managed by thrombectomy (n = 2), thrombectomy followed by retransplantation (n = 3), retransplantation (n = 2), and revision (n = 2). Nonsurgical approaches included angioplasty (n = 6) and observation (n = 4). The overall survival rate was 65% (13 of 20). HAT is the most serious complication following liver transplantation; it may lead to death. Timely retransplantation may salvage some patients, and angioplasty is useful in the management of vascular stenoses. PMID- 11890338 TI - Successful partial transplant pancreatectomy with end-to-side duct-to-ureter anastomosis: salvage of a pancreatic allograft. AB - Segmental pancreatic transplantation has been abandoned because of the high incidence of technical complications. We report the first case in the literature of the salvage of a partially ischemic pancreatic allograft. The procedure consisted of resecting the head of the pancreas and draining the residual segment to the ureter with a duct-to-ureter end-to-side anastomosis. The postoperative course was uneventful, and 15 months after surgery graft function is satisfactory with a urinary amylase level of 5000 U/h. The duct-to-ureter drainage technique should be part of every transplant surgeon's repertoire, because in emergency situations like the one described, it can be used to save a pancreatic allograft. PMID- 11890340 TI - An unusual case of iatrogenic severe hypernatremia. PMID- 11890339 TI - Total laryngectomy: pre- and intrasurgical variables of infection risk. AB - Postoperative infection has influence on costs, quality of life, and outcome of the disease. It is suspected that post-total laryngectomy infections have increased in frequency and seriousness, because of the failure of the preservation protocol or the previous radiotherapy, making rescue surgery necessary. The objective of this study was to develop a predictive model of infection based on the pre- and intrasurgical variables considered risky. One hundred fifty five patients with E III-IV laryngeal cancer, with 24.8:1 male to female ratio (mean age, 58 years) who underwent total laryngectomy were evaluated for uni- and multivariate analysis of age, sex, histological grade, primary or recurrent disease, tobacco, alcohol, diabetes, tuberculosis/chronic emphysema, red and white cell counts, erythrosedimentation rate (ESR), albumin, chemotherapy, neck radiotherapy and/or previous surgery, confinement days, type and time of surgery, which were factors in the infection event. A predictive model of infection was developed and included albuminemia (<3.5 g%), >1 liter of alcohol daily, and exclusive surgery of the primary. The sensitivity was 90.5% and the specificity 68%. The variance reached 29.6%. The causes of infection were multiple, having analyzed only 30% of them. However, the resulting model was classified correctly in 83.2% of cases. A careful preoperative assessment, an adjusted planning of the surgery, an appropriate use of antibiotics, and a meticulous operative technique are needed to prevent infection. PMID- 11890341 TI - Gastrectomy with D2 lymph node dissection in gastric cancer: a retrospective study at a single institution. AB - The purpose of this study was to clarify the clinicopathological characteristics of gastric cancer with lymph node (LN) dissection and the significance of D2 dissection by investigating surgical techniques and prognosis. Three hundred ninety patients with early cancer and 310 with advanced cancer underwent gastrectomy with D1 or D2 dissection, based on the presence or absence of LN metastasis determined pre- and intraoperatively. LN metastasis occurred in 10.5% of early gastric cancer patients, and several cases of advanced cancer were found to have N2 or more advanced metastasis. The pre- and intraoperative macroscopic findings accorded with histological grade of LN metastasis in 69.5% of early cancers and in 56.5% of advanced cancer patients. The false negativity rate was 6.8% in early cancer, 19.4% in advanced cancer, and 8.4% as a whole. Death was operation-related in only two cases and the operative mortality rate was low (0.29%). The 5-year survival rates in early and advanced gastric cancer were 95.8% and 67.6% in the D1 groups, respectively, and 100% and 89.5% in the D2 groups, respectively. Survival was better in the D2 groups than in the D1 groups (P < 0.0001 for early cancer, P = 0.0279 for advanced cancer). D2 dissection should be conducted positively for patients with LN metastasis. PMID- 11890342 TI - Mesenteric actinomycosis with retroperitoneal involvement. AB - Mesenteric or retroperitoneal actinomycosis is an extremely rare disease. The international databases have revealed only 10 cases affecting the mesenterium and another 52 cases affecting the retroperitoneum. We report a 78-year-old female who was admitted with complaints of abdominal pain. Laboratory examination revealed anemia and the clinical examination revealed an irregular mass in the abdomen. Ultrasound and computed tomography (CT) scans showed a solid mass in the mesenteric-retroperitoneal region. Biopsy of the nonresectable mass revealed the presence of chronic inflammation in the mesenteric area with Actinomyces colonies. The patient was treated with oral amoxicillin, 500 mg every 6 hours for 6 months. The symptoms disappeared, but the mesenteric-retroperitoneal mass remains, but smaller in size. Based on the review of the literature and the case reported here, we conclude that mesenteric-retroperitoneal actinomycosis is difficult to diagnose by means of noninvasive techniques as it can masquerade as a malignant process. An accurate diagnosis is always obtained in a histological or microbiological examination, often requiring surgical intervention. Treatment with penicillin has proven to be effective. PMID- 11890343 TI - Descending necrotizing mediastinitis: a case report and review of the literature. AB - Descending necrotizing mediastinitis (DNM) is a highly fatal disease and as infection spreads along deep cervical planes into the mediastinum, widespread cellulitis, necrosis, abscess formation, and sepsis may occur. Early diagnosis is crucial for starting aggressive treatment without delay. Cervicothoracic computed tomography (CT) scanning may be useful for early diagnosis and preoperative evaluation of the surgical approach. Optimal treatment includes broad-spectrum antimicrobial therapy and extensive cervicomediastinal and transthoracic drainage. Clamshell incision provides an excellent exposure of both thoracic cavities and all mediastinal structures with minimal morbidity. We report here a fatal case of DNM with bilateral empyema and purulent pericarditis due to an odontogenic abscess with a brief review of the literature. PMID- 11890344 TI - Pyogenic liver abscess after hepatobiliary and pancreatic surgery. AB - From 1984 to 1998, a total of 2158 patients underwent hepatobiliary and pancreatic surgery, and 12 patients developed liver abscess after hepatobiliary and pancreatic surgery; thus, the incidence of liver abscess was 0.6%. The main reasons for liver abscess were anastomotic stricture in 5 patients, obstruction of percutaneous transhepatic biliary drainage (PTBD) tube in 3 patients, and portal vein and hepatic artery obstruction due to intraoperative radiation in 1 patient, transportal chemotherapy in 1 patient, chemo-lipiodolization in 1 patient, and unknown in 1 patient. Ten of the 12 patients initially underwent percutaneous transhepatic abscess drainage of whom 2 patients subsequently received surgical drainage. The other 2 patients were treated with antibiotics only. Eight of the 12 patients were cured, but 4 patients died. The reasons for death were sepsis in 3 patients and liver failure due to portal vein and hepatic artery obstruction in 1 patient. Our results indicate that liver abscess should be taken into consideration for patients with risk factors. PMID- 11890345 TI - Cardiac herniation mimics cardiac tamponade in blunt trauma. Must early resuscitative thoracotomy be done? AB - Blunt rupture of the pericardium is a rare injury. Strangulated cardiac hernia following blunt trauma is one cause of reversible cardiac arrest. Traumatic pericardial tears usually have delayed diagnoses and carry high mortality rates (64%). Clinical signs mimic cardiac tamponade during the primary survey. We report here two cases of blunt trauma. Both patients arrived alive in the emergency room and presented signs of cardiac tamponade caused by pericardial rupture. PMID- 11890346 TI - Treatment of patients with gastric cancer and duodenal invasion. AB - We retrospectively examined clinicopathologic features of gastric cancer with duodenal invasion to clarify the effect of surgical treatment that include pancreaticoduodenectomy (PD). Among 2504 patients with gastric cancer, 69 (2.8%) who had gastric cancer and duodenal invasion resected by surgical treatment were investigated. The mode of the duodenal invasion was grouped into three categories: mucosal type, submucosal type, and nodal type. Mucosal type is invasion of the duodenal mucosal layer, submucosal type is invasion of the submucosal layer or deeper, and nodal type is invasion from nodal metastatic lesions around the pancreatic head. The 5-year survival rates of curative PD and curative gastrectomy were 37.3% and 33.8%, respectively. Despite the incidence of adjacent tissue infiltration and significantly higher duodenal invasion average length in cases with PD than in cases with gastrectomy, there was no significant difference in the survival curves. However, the prognoses of the cases with nodal type invasion were significantly poorer, and all these patients died within 2 years, regardless of whether curative PD had been performed. Curative PD improves the prognosis of cases with long duodenal invasion or pancreas infiltration except for nodal-type duodenal invasion. PMID- 11890347 TI - Overview: antifungal combination therapy. AB - We have yet to realize the clinical benefits of combination antifungal therapy, but the literature is beginning to show the potential positive and negative aspects of this approach. Detailed laboratory-based study of each new antifungal compound will be required in order to define the interactions between the different drugs. Most important in this discussion is that a determination of the most appropriate experimental design and parameters to reflect the outcomes in the treatment of human mycoses is required. That will be the challenge for the next several years. In the meantime, combination antifungal therapy should be approached carefully and be initiated on a case-by-case basis following consideration of all the options available for the patient. PMID- 11890348 TI - Overview: non-fumigatus species of Aspergillus: perspectives on emerging pathogens in immunocompromised hosts. PMID- 11890350 TI - The treatment of pulmonary aspergilloma. AB - Aspergillomas are fungal balls within lung cavities. The natural history of patients affected is variable. Hemoptysis is a dangerous sequela. Factors associated with a poor prognosis have been defined and therapy is difficult because of the lack of a blood supply. Randomized trials are lacking. Surgical treatment is definitive but many patients are ineligible. Percutaneous therapy and bronchial artery embolization is appropriate for some patients and itraconazole has produced favorable results in several studies. PMID- 11890349 TI - Anti-toxoplasmosis drugs. AB - The drugs most commonly used for treatment of toxoplasmosis combine folate inhibitors that frequently cause undesirable side effects. There is an urgent need for new drugs or drug combinations that are able to eradicate the tissue cysts responsible for relapses, particularly for patients intolerant to folate inhibitors. Although promising for its activity against cysts, atovaquone is still limited by its variable intestinal absorption. The recent demonstration that the apicoplast of Toxoplasma gondii could be a possible unique drug target for antibiotics such as macrolides and fluoroquinolones represented an important breakthrough in this area. PMID- 11890351 TI - Overview: Nod2, cause of, or contributor to, Crohn's disease. PMID- 11890352 TI - The role of the gut lymphoid tissue in induction of oral tolerance. AB - The gut-associated lymphoid tissue (GALT) maintains a balance between immunological tolerance to dietary proteins and induction of active immune responses to pathogenic microorganisms. The oral administration of soluble protein antigens induces a state of systemic immunological unresponsiveness specific to the fed protein, termed oral tolerance. The two major mechanisms to explain oral tolerance are anergy/deletion of autoreactive lymphocytes and active suppression. This review will discuss the mechanisms of therapeutic oral tolerance in relation to events occurring at the site of antigen entry. PMID- 11890353 TI - Anti-inflammatory peptides and proteins in inflammatory bowel disease. AB - Current therapy of inflammatory bowel disease, ie, ulcerative colitis and Crohn's disease, is neither sufficient nor disease-modifying. Long-term treatment with non-specific antiinflammatory drugs aminosalicylates, corticosteroids and immunosuppressants is often accompanied with undesirable and potentially serious side effects. Novel biologically-driven therapies are targeted to specific pathophysiological processes, offering the potential for better treatment outcomes. Among other antiinflammatory peptides and proteins, monoclonal antibodies directed against TNFalpha and adhesion molecule alpha4beta7 integrin, recombinant anti-inflammatory cytokines IL-10 and IL-11, as well as colony stimulating factors and peptide growth factors, are in the most advanced stages of clinical development for IBD. PMID- 11890354 TI - Oprelvekin. Genetics Institute. AB - Genetics Institute has developed and launched oprelvekin (rhIL-11; Neumega), a recombinant form of human IL-11. In November 1997, the FDA cleared oprelvekin for the prevention of severe thrombocytopenia and the reduction of the need for platelet transfusions following myelosuppressive chemotherapy in susceptible patients with non-myeloid malignancies 12703021. The product was launched at the end of 1997 [312556]. By December 1999, phase III trials for Crohn's disease (CD) were underway [363007]. Genetics Institute had commenced a 150-patient phase II trial for mild-to-moderate CD and mucositis and the company planned to file regulatory procedures for the indication of CD in 1999 [271210]. An oral formulation for this indication has been developed. Oprelvekin is also undergoing phase I clinical trials for colitis [396157], phase II clinical trials for rheumatoid arthritis [413835] and clinical trials for psoriasis [299644]. In March 1997, Wyeth-Ayerst became the licensee for Europe, Africa, Latin America and Asia (with the exception of Japan). Genetics Institute holds marketing rights for North America [239273]. In Japan, oprelvekin is being developed by Genetics Institute and Yamanouchi; phase III trials have commenced [295049] and were ongoing in May 2001 [411763]. In April 1996, analysts at Yamaichi estimated launch in 2001 and maximum annual sales of over yen 10 billion [215896]. In January 1998, Morgan Stanley Dean Witter predicted Yamanouchi's share of sales to be yen 1 billion in 2001, rising to yen 2 billion in 2002 [315458]. Sales of oprelvekin were US $34 million for Genetics institute in fiscal 2000 while, in July 2001, Credit Suisse First Boston estimated that this figure will be US $30 million and US $34 million in 2001 and 2002, respectively [416883]. PMID- 11890355 TI - Alicaforsen. Isis Pharmaceuticals. AB - Alicaforsen (ISIS-2302) is an RNase H-dependent antisense inhibitor of the intercellular adhesion molecule ICAM-1 under development by Isis Pharmaceuticals, for the potential treatment of a variety of inflammatory disorders [175741]. As of April 1997 it was in phase III trials for Crohn's disease (CD); however, the trial failed and, in December 1999, the company suspended development for this indication [352801]. In October 2000, the company re-initiated development in CD [384820] and new phase III trials had begin by May 2001 [409704]. In August 2000, phase II studies of alicaforsen in an enema formulation for ulcerative colitis and a topical formulation for psoriasis were ongoing [378715]. Development of the compound for the potential treatment of rheumatoid arthritis (RA) was discontinued in 1999 [347579]. By the end of 1998, alicaforsen was in phase II trials for kidney transplant rejection. At this time, these trials were expected to finish in mid-1999 [343460]. However, they were ongoing in September 1999, although no further development has been reported for this indication since that time [338672]. In February 1995, Isis Pharmaceuticals and Boehringer Ingelheim (BI) signed a collaborative agreement on cell adhesion inhibitors, including alicaforsen [174111]. By early 1999, Isis and BI were to decide on the next developmental step for alicaforsen following further analyses of its performance against CD [292915], [315439]. Their joint development agreement was terminated in 1999; Isis regained rights to the product and by September 1999 was in talks to license alicaforsen to another partner for CD [338672]. In June 2000, Cytogenix entered into a sponsored research agreement with Baylor College of Medicine at the Texas Medical Center Houston for the use of its ssDNA expression system for the development of antisense strategies directed against intercellular adhesion molecules for the purpose of reducing lung inflammation and injury in disease states and conditions [369677]. US-05514788, and other patents, cover antisense cell adhesion molecule inhibitors [212289], [234792]. PMID- 11890356 TI - Pharmacological treatment of portal hypertension. AB - In liver cirrhosis, increased resistance to portal blood flow is the primary factor in the pathophysiology of portal hypertension. The recognition of a dynamic component in hepatic resistance due to the active-reversible contraction of different elements of the portohepatic bed, has led to the active development of hepatic vasodilators. On the other hand, a significant increase in portal blood flow caused by arteriolar splanchnic vasodilation and hyperkinetic circulation, aggravates portal hypertension and provides the rational for the use of splanchnic vasoconstrictors, such as beta-blockers, vasopressin derivatives and somatostatin and its analogs. This review covers current developments in the treatment of portal hypertension. PMID- 11890357 TI - Omapatrilat. Bristol-Myers Squibb. AB - Bristol-Myers Squibb (BMS) is developing the vasopeptidase inihibitor, omapatrilat, a dual inhibitor of angiotensin converting enzyme (ACE) and neutral endopeptidase (NEP), for the potential treatment of cardiovascular diseases such as hypertension and heart failure [306287]. An NDA for the use of omapatrilat in hypertension was filed with the FDA and the regulatory authorities in the EU in December 1999 [351207], [353287]. In April 2000, BMS voluntarily withdrew the NDA in response to questions raised by the FDA regarding the comparative incidence and severity of an infrequent side effect (angioedema) reported within the NDA database. Prospective controlled clinical studies in patients with hypertension and heart failure were to continue. In May 2001, BMS reported that its blinded omapatrilat hypertension study was continuing and, pending supportive results from a data analysis anticipated in late summer/early autumn 2001, the company expected to refile an NDA with the FDA [409203]. In July 2000, BMS reported that it planned to conduct a multinational, 25,000 patient study (OCTAVE - Omapatrilat Cardiovascular Treatment Assessment Versus Enalapril) to compare the efficacy and safety of omapatrilat against enalapril in the treatment of hypertension [374909]. The OCTAVE trial was expected to generate data by mid-2001, which could allow for a launch by early 2002 [380280]. Phase III trials for hypertension had commenced by January 1998 [273646]. In January 2001, Merrill Lynch expected BMS to refile its NDA with the FDA in the second half of 2001 [395423]. In February 2001, Credit Suisse First Boston made a similar prediction, adding that it believed BMS would launch the drug in late 2002 or early 2003. The analysts also predicted peak sales for the drug of $585 million in 2005 [399484]. In May 2001, Merrill Lynch estimated sales of $1.8 billion in 2005 [411811]. PMID- 11890358 TI - SR-121463. Sanofi-Synthelabo. AB - Sanofi-Synthelabo (formerly Sanofi) is developing SR-121463, a vasopressin V, receptor antagonist, as a potential treatment for cardiovascular indications such as congestive heart failure (CHF) and hypertension [330073], [341858]. By September 2001, it had entered phase IIa trials for these indications [421268]. SR-121463 was in phase I clinical trials for CHF and hypertension in June 2001 [359231], [413342]. It was also being evaluated for the potential treatment of glaucoma but its development has been discontinued for this indication [367094]. In October 1999, Lehman Brothers predicted a 5% chance of the compound reaching market, with a launch anticipated in 2004 and potential peak sales of $100 million in 2012 [346267]. PMID- 11890360 TI - Cilansetron. Solvay. AB - Cilansetron is a 5-HT3 antagonist tinder development by Solvay Pharmaceuticals as a potential treatment for irritable bowel syndrome (IBS). The compound targets non-constipated men and women with IBS. Cilansetron is being evaluated in phase III trials, but up to now, only scarce information has been made available and most publications have appeared in abstract form only. In July 2001, it was reported that regulatory submissions were expected in 2003 [416185]. Also in July 2001, after discussion with the FDA, Solvay initiated a revised phase III program in diarrhea-predominant IBS and signed a five-year 'preferred-provider' clinical services agreement with Quintiles Transnational to conduct the trial. At this time, Solvay was also seeking marketing partners for cilansetron [416185]. By October 1999, Solvay was treating cilansetron as one of its main priorities, as it represented a novel class of compound [342434]. Solvay has predicted peak sales of euro 100 m to euro 1000 m [420654]. PMID- 11890359 TI - Overview: H3 histamine receptor isoforms: new therapeutic targets in the CNS? PMID- 11890361 TI - TAK-637. Takeda. AB - Abbott and Takeda are developing TAK-637, an orally active NK1 antagonist, for the potential treatment of urinary incontinence, depression, irritable bowel syndrome and pollakiuria. By November 1999, it was in phase II trials in Europe and phase I in Japan and the US for urinary incontinence [348496], [350686]. By October 2000, phase II trials had been initiated in the US for urinary incontinence, depression and IBS [381167], [386950], [419868], and in May 2001, these were scheduled to finish in 2002 [412024]. PMID- 11890362 TI - Eziopitant. Pfizer. AB - Pfizer is developing ezlopitant, a neurokinin-1 antagonist, for the potential treatment of irritable bowel syndrome (IBS). The compound had undergone phase II trials in the US and Europe, and phase I in Japan for treatment of chemotherapy induced emesis [290988], [320737], [329187]. A phase II, double-blind, randomized study was performed to assess the safety and efficacy of ezlopitant for the control of cisplatin-induced emesis. Treatment was well tolerated [290988]. Although the compound effectively controls emesis, it is less effective in controlling nausea, and development has been discontinued for the emesis indication [347367]. Ezlopitant has undergone a pilot study in 14 IBS patients [367631]. PMID- 11890363 TI - Drug properties of second-generation antisense oligonucleotides: how do they measure up to their predecessors? AB - Antisense technology has progressed beyond the point of using only phosphorothioate oligodeoxynucleotides as therapeutic agents to looking at antisense molecules that contain additional chemical modifications as the next generation of therapeutic agents. These modifications are intended to improve the overall therapeutic properties by increasing potency, optimizing pharmacokinetic properties and improving the safety profile. This review will focus on the non clinical pharmacokinetic and safety properties of 2'-O-methoxyethyl-modified oligonucleotides. Implications on the convenience and safe use of these compounds as therapeutic agents will be discussed. PMID- 11890364 TI - Cellular uptake of antisense oligonucleotides. AB - Antisense oligonucleotides (AS ONs) selectively bind to the target mRNA and prevent its translation into the corresponding protein. Various tissue culture studies demonstrated that AS ONs enter into cells via the receptor-mediated endocytosis pathway. There are many different types of receptors, and their characteristics and expression vary with cell types. In this review, we will discuss the characteristics of the various receptors that have been isolated in vitro. We will also discuss the uptake and the bioavailability of AS ONs after being administered in vivo. PMID- 11890365 TI - ISIS-3521. Isis Pharmaceuticals. AB - ISIS-3521 is a 20-mer antisense phosphorothioate oligonucleotide PKCa expression inhibitor, under development by Isis (formerly in collaboration with Novartis) for the potential treatment of solid tumors that are refractory to, or recurrent with, standard treatment regimens [175741]. In November 1999, Novartis announced that it would end its codevelopment of ISIS-3521 [348221], [348222]. In August 2001, Eli Lilly in-licensed ISIS-3521 [420062]. In October 2000, phase III trials of ISIS-3521, in combination with carboplatin and paclitaxel, were initiated for the treatment of non-small cell lung cancer (NSCLC) [386128]. The FDA granted ISIS-3521 Fast Track review status for NSCLC in November 2000 [388930]. In April 2001, Bear Sterns & Co predicted US approval of ISIS-3521 in 2002 [411081]. In August 2001, Eli Lilly and Isis entered into a four-year strategic alliance that includes ISIS-3521. For the license of ISIS-3521, Isis will receive $25 million in upfront fees and will be reimbursed for remaining phase III development and registration costs [420062]. PMID- 11890366 TI - GTI-2040. Lorus Therapeutics. AB - Loris Therapeutics (formerly GeneSense Therapeutics) is developing the antisense oligonucleotide GTI-2040, directed against the R2 component of ribonucleotide reductase, for the potential treatment of cancer [348194]. It is in phase I/II trials [353796] and Lorus had anticipated phase II trials would be initiated in July 2001. By August 2001, GTI-2040 was undergoing a phase II trial as a monotherapy for the potential treatment of renal cell carcinoma, and was about to enter a phase II combination study for this indication with capecitabine (Hoffmann-La Roche). At this time, the company was also planning a phase II trial to study the drug's potential in the treatment of colorectal cancer [418739]. GTI 2040 has been tested in nine different tumor models, including tumors derived from colon, liver, lung, breast, kidney and ovary. Depending on the tumor model, significant inhibition of tumor growth, disease stabilization and dramatic tumor regressions was observed [347683]. Lorus filed an IND to commence phase I/II trials with GTI-2040 in the US in November 1999 [347683], and received approval for the trials in December 1999 [349623]. As of January 2000, these trials had commenced at the University of Chicago Cancer Research Center; it was reported in February 2000 that dosing to date had been well tolerated with no apparent safety concerns [357449]. Lorus has entered into a strategic supply alliance with Proligo to provide the higher volumes of drug product required for the planned multiple phase II trials [385976]. In February 1998, Genesense (now Lorus) received patent WO-09805769. Loris also received a patent (subsequently identified as WO-00047733) from the USPTO in January 2000, entitled 'Antitumor antisense sequences directed against components of ribonucleotide reductase' covering the design and use of unique antisense anticancer drugs, including GTI 2040 and GTI-2501 [353538]. PMID- 11890367 TI - GTI-2501. Lorus Therapeutics. AB - Lorus Therapeutics (formerly GeneSense Technologies) is developing GTI-2501, directed against the R1 component of ribonucleotide reductase, for the potential treatment of cancer [348928]. In November 2000, the company filed an IND with the FDA in preparation to begin a phase I trial in the US in the first quarter of 2001 [389975], [396537], [397960]. In March 2001, the FDA approved the IND application [402350], and in June 2001, a phase I, dose-escalating trial in patients with lymphomas or solid tumors which have not responded to standard therapy commenced in the US [412225]. The company expected the phase I trial of GTI-2501 to be completed within 12 to 15 months [420582]. In November 1999, the company reported that in preclinical trials, GTI-2501 was an effective anticancer agent when tested in standard mouse models bearing a variety of different human cancer lines including tumor cells derived from lung, breast, colon, kidney, ovary pancreas and skin cancers [348928]. Lorus received a patent from the US PTO in January 2000, covering the design and use of unique anitisense anticancer drugs, including GTI-2501 and GTI-2040 [353538]. In June 2000, the USPTO allowed a patent to specifically protect GTI-2501 [370810]. PMID- 11890369 TI - Plasma C-reactive protein predicts left ventricular remodeling and function after a first acute anterior wall myocardial infarction treated with coronary angioplasty: comparison with brain natriuretic peptide. AB - BACKGROUND: C-reactive protein (CRP) directly participates in the myocardial injury of acute myocardial infarction (MI). Although high plasma CRP levels in the acute phase strongly indicate a poor early clinical outcome of patients with MI, the impact of CRP levels on late left ventricular (LV) function and remodeling, which are closely associated with long-term prognosis, remains unknown. HYPOTHESIS: Acute plasma CRP levels may predict late LV function and remodeling after MI. METHODS: We prospectively studied 12 consecutive patients with a first acute anterior MI recanalized by angioplasty. We measured plasma CRP levels on Days 0, 1, 2, 3, 4, and 7, and calculated the area under the curve (AUC). We also measured plasma brain natriuretic peptide (BNP) levels on Day 3 as the referential indicator of LV dysfunction and late LV remodeling. Late LV indices were independently assessed on a left ventriculogram obtained at 5.3 months to estimate the extent of LV remodeling. RESULTS: Plasma CRP reached its peak at Day 2.8 (8.68+/-4.57 mg/dl). On linear regression analysis, the AUC of CRP (35.21+/-19.33 mg/dl x day) correlated positively with BNP (316.5+/-418.6 pg/ml) (r = 0.646, p = 0.023). The AUC of CRP, peak CRP, and BNP correlated significantly with late LV indices. Among these, the AUC of CRP showed the best correlation with end-diastolic volume index (r = 0.765, p = 0.004), end-systolic volume index (r = 0.907, p < 0.001), and ejection fraction (r = -0.862, p < 0.001). CONCLUSIONS: Patients with high plasma CRP levels may be at risk for late LV dysfunction and remodeling; theoretically, their long-term prognosis may be poor. Measuring plasma CRP levels may provide valuable information for long-term risk stratification after MI. PMID- 11890368 TI - Correlation between myocardial perfusion abnormalities detected with intermittent imaging using intravenous perfluorocarbon microbubbles and radioisotope imaging during high-dose dipyridamole stress echo. AB - BACKGROUND: The clinical accuracy of myocardial contrast echocardiography (MCE) using intermittent harmonic imaging and intravenous perfluorocarbon containing microbubbles during dipyridamole stress has not been evaluated in a multicenter setting. HYPOTHESIS: The accuracy of dipyridamole stress contrast echo in the detection of coronary artery disease (CAD) using myocardial perfusion images is high in comparison with technetium-99 (99Tc) sestamibi single-photon emission computed tomography (MIBI SPECT) and increases the accuracy of wall motion data. METHODS: In 68 consecutive nonselected patients (46 men; mean age 66 years) from three different institutions in two countries. dipyridamole stress echo and SPECT with 99mTc MIBI were compared. Continuous intravenous (IV) infusion of perfluorocarbon exposed sonicated dextrose albumin (PESDA) (2-5 cc/min) was administered for baseline myocardial perfusion using triggered harmonic end systolic frames. Real-time digitized images were used for wall motion analysis. Dipyridamole was then injected in two steps: (1) 0.56 mg/kg for 3 min, (2) 0.28 mg/kg for 1 min, if the first step was negative for an inducible wall motion abnormality. After dipyridamole injection, myocardial contrast enhancement and wall motion were analyzed again by the same methodology. RESULTS: There were 35 patients with perfusion defects by SPECT. Wall motion was abnormal in 22, while MCE was abnormal in 32. Wall motion and MCE each had one false positive. The proportion of correctly assigned patients was significantly better with MCE than with wall motion (p = 0.03; chi square test). CONCLUSIONS: Myocardial contrast echocardiography, using intermittent harmonic imaging and intravenous perfluorocarbon containing microbubbles, is a very effective method for detecting coronary artery disease during dipyridamole stress echo. PMID- 11890370 TI - A comparison of treadmill scores to diagnose coronary artery disease. AB - BACKGROUND: Recently, several treadmill scores have been proposed as means for improving the diagnostic accuracy of the exercise treadmill test (ETT). Questions remain regarding the diagnostic accuracy of treadmill scores when applied to a different patient population than that from which they were derived; furthermore, many treadmill scores have not been compared with one another in the same population. HYPOTHESIS: The diagnostic accuracy of treadmill scores may not be the same. METHODS: A retrospective analysis of data collected prospectively was performed on consecutive patients referred for evaluation of chest pain. All patients underwent a standard ETT followed by coronary angiography. Using angiographic evidence of coronary artery disease (CAD) as a reference, the area under the curve (AUC) of receiver operator characteristic (ROC) plots of the ST response alone, the Duke Treadmill Score (DTS), the Morise score, the Detrano score, the VA score, and a Consensus score consisting of the Morise, Detrano, and VA scores together were calculated and compared. The predictive accuracies of the DTS and the Consensus score to stratify patients for the likelihood of CAD were calculated and compared. RESULTS: In all, 1,282 patients without a prior myocardial infarction had an ETT and coronary angiography. The AUC (+/- standard error) was 0.67+/-0.01 for the ST response, 0.73+/-0.01 for DTS, 0.76+/-0.01 for Detrano score, 0.77+/-0.01 for Morise score, 0.78+/-0.01 for VA score, and 0.78+/ 0.01 for Consensus score. The AUC for each treadmill score was significantly higher (z-score > 1.96) than for the ST response alone. The AUC of DTS was significantly lower than all other treadmill scores (z-score > 1.96). The predictive accuracy (+/-95% confidence interval) of the DTS to risk stratify patients into high and low likelihood for CAD was 71 (65-77)%, versus 80 (74-86)% for the Consensus score (p < 0.0001). CONCLUSION: In this population, the DTS remains useful for diagnosing CAD and stratifying for the likelihood of CAD, although it is less accurate than other treadmill scores. PMID- 11890371 TI - Adaptive mechanisms of left ventricular diastolic function to the physiologic load of pregnancy. AB - BACKGROUND: Pregnancy is associated with marked alteration in cardiovascular hemodynamics. Recent reports have characterized the effects on cardiac systolic function. Little has been written on the influences of loading conditions on Doppler measures of diastolic function during pregnancy. HYPOTHESIS: Stage of pregnancy has an impact on Doppler indices of diastolic function independent of loading conditions, systolic function, and heart rate. METHOD: Thirty healthy women were prospectively evaluated by serial echocardiography and Doppler examinations at six time periods: 10-12, 18-20, 28-30, 36-38 weeks gestation, 2-4 and 12-14 weeks postpartum. The related effects on indices of diastolic function and its interaction with load, heart rate, mass, and systolic function were determined. RESULTS: Compared with the nonpregnant state, early (E) velocity increased (0.7+/-0.1-0.9+/-0.1 m/s, p = 0.0001), peaking at 18 weeks and returning to normal levels during late pregnancy. Atrial phase (A) velocity peaked at 18 weeks (0.48+/-0.12-0.60+/-0.13 m/s, p = 0.0001), remaining high throughout the rest of pregnancy. Consequently, the EWA ratio fell significantly during late pregnancy, from 1.9+/-0.4 to 1.4+/-0.3 (p = 0.02). In addition, mean acceleration was significantly increased in early pregnancy with a peak at 18 weeks (7.4+/-1.3 m/s2), returning to nonpregnant level at term (5.7+/-1.4 m/s2, p = 0.0001). Generalized estimating equation using multivariate regression analysis demonstrated that rising heart rate and stroke volume index had an independent effect on A velocity, and that contractility and preload had an independent effect on E velocity. Pregnancy itself had an independent influence on early filling, not explained by the other parameters. CONCLUSIONS: During normal pregnancy, there is a reversible shift in transmitral flow velocities from early to late filling with a decrease in acceleration, consistent with an increase in ventricular compliance. Changes in heart rate, preload, and contractility, as well as stage of pregnancy influence this alteration. PMID- 11890372 TI - Differential diagnosis of "dysphagia" in an elderly woman. PMID- 11890373 TI - Myocardial infarction following the combined recreational use of Viagra and cannabis. AB - Sildenafil citrate (Viagra, Pfizer, Inc., New York, N.Y.) is widely prescribed as a treatment for male erectile dysfunction. It is metabolized predominantly by the cytochrome P450 3A4 hepatic microsomal isoenzyme and effects can, therefore, be potentiated by such inhibitors. The vasodilatory effects of Viagra necessitate caution in its use in patients with cardiovascular disease and it is contraindicated in patients receiving nitrates. Previous literature has drawn attention to Viagra use and myocardial infarction. This paper reports the case of a young man who presented with a myocardial infarction after taking Viagra in combination with cannabis, a known inhibitor of the cytochrome P450 3A4 isoenzyme. PMID- 11890374 TI - Realdo Colombo. PMID- 11890375 TI - Evolution of NCEP guidelines: ATP1-ATPIII risk estimation for coronary heart disease in 2002. National Cholesterol Education Program. PMID- 11890376 TI - Coronary flow velocity reserve in hypertensive patients with left ventricular systolic dysfunction. AB - BACKGROUND: Hypertensive microvascular disease is speculated to be a limiting factor for the ability of left ventricular (LV) hypertrophy to maintain LV systolic function in systemic hypertension. The role of coronary reserve, which may be affected by microvascular disease, remains uncertain in the pathophysiology of hypertensive heart disease. HYPOTHESIS: A progressive impairment of coronary flow velocity reserve (CFVR) according to the presence and severity of LV systolic dysfunction is anticipated to occur in hypertension. METHODS: According to the absence or presence of LV dysfunction (LV fractional shortening - FS% < 30), two groups of hypertensive patients were investigated: HP1 (n = 9, FS% = 36+/-6) and HP2 (n = 13, FS% = 18+/-6). Eight normal subjects (NL) served as controls (LVFS% = 35+/-3). Doppler blood flow velocity was obtained from the left anterior descending coronary artery using transesophageal echocardiography before, and during 6-min continuous adenosine infusion (140 microg x kg(-1) x min(-1) intravenous). The CFVR was calculated as the ratio of maximal to baseline peak diastolic flow velocities. RESULTS: The comparison among NL, HP1, and HP2 groups showed statistically different (p < 0.05) mass index (101+/-18, 172+/-46, and 257+/-54 g x m(-2)), end-systolic wall stress (76.9+/ 14.4, 78.4+/-23.9, and 174.5+/-43.0 10(3) x dyn x cm(-2)), and CFVR (3.5+/-0.6, 3.2+/-0.4, and 2.6+/-0.8), respectively. The CFVR correlated significantly and directly with LVFS% (r = 0.40) and correlated inversely with both mass index (r = -0.54) and end-systolic stress (r = -0.40). CONCLUSIONS: These results indicate that CFVR impairment is weakly related to LV dysfunction in hypertension. PMID- 11890377 TI - 'We will look for health': the Osteopathic Center for Children. PMID- 11890378 TI - How healing happens: exploring the nonlocal gap. PMID- 11890379 TI - Oculomotor nerve palsy treated with acupuncture. PMID- 11890380 TI - Homeopathy deserves more of this journal's attention. PMID- 11890381 TI - 'State of Center,' international research top agenda of 10th NACCAM meeting. National Advisory Council for Complementary and Alternative Medicine. PMID- 11890382 TI - Directions for research in complementary medicine and bioterrorism. PMID- 11890383 TI - Etiology and treatment of uterine fibroids. PMID- 11890384 TI - Complementary medicine treatment of uterine fibroids: a pilot study. AB - CONTEXT: Women are seeking alternatives to medications and surgeryfor the treatment of uterine fibroids; therefore, it is important to know whether these therapies work and are cost-effective. OBJECTIVE: To report on the treatment of women with uterine fibroids using a suite of nonpharmacological and nonsurgical therapies and to compare this alternative treatment to a matched sample of similar women using standard medical approaches. DESIGN: Matched comparison to standard-of-care control group. SETTING: Academic family practice department and clinic, including a center for complementary and alternative medicine. PARTICIPANTS: Thirty-seven menstruating women, aged 24 to 45 years, with palpable uterine fibroids, and their matched controls, who were enrolled in conventional treatment. INTERVENTION: The treatment program consisted of weekly traditional Chinese medicine, body therapy (somatic therapy, bodywork), and guided imagery. Treatment lasted as long as 6 months. MAIN OUTCOME MEASURES: Changes in fibroid size, changes in bothersome symptoms, and patient satisfaction. RESULTS: Fibroids shrank or stopped growing in 22 patients among the treatment group and 3 among the comparison group (P <.01). Cost of care averaged $3800 per patient in the treatment group and was significantly greater than costs for the comparison group for the study period. Bothersome symptoms of fibroids responded equally well to pharmacological or nonpharmacological therapies. All measures of patient satisfaction were significantly higher among the treatment group compared to those receiving conventional care. CONCLUSIONS: As reported previously in the Chinese medical literature, alternatives exist to pharmacological and surgical methods for treating uterine fibroids, though they are not inexpensive. PMID- 11890385 TI - Experience of a Reiki session. AB - CONTEXT: Touch therapies, including Reiki, are increasingly popular complementary therapies. Previous studies of touch therapies have yielded equivocal findings. OBJECTIVE: Exploring the experiences of Reiki recipients contributes to understanding the popularity of touch therapies and possibly elucidates variables for future studies. DESIGN: Descriptive study with quantitative and qualitative data. This report focuses on qualitative interview data. Thematic analysis was used to discern patterns in the experience. SETTING: All Reiki treatments were given in a sound proof windowless room by one Reiki master. Audiotaped interviews were conducted immediately after the treatment in a quiet room adjoining the treatment room. PARTICIPANTS: Generally healthy volunteers (N=23) who were naive to Reiki. INTERVENTION: Standardized, 30-minute Reiki session. MAIN OUTCOME MEASURES: Interview data supported by quantitative data. RESULTS: Participants described a liminal state of awareness in which sensate and symbolic phenomena were experienced in a paradoxical way. Liminality was apparent in participants' orientation to time, place, environment, and self Paradox also was seen in participants' symbolic experiences of internal feelings, cognitive experience, and external experience of relationship to the Reiki master. CONCLUSIONS: Liminal states and paradoxical experiences that occur in ritual healing are related to the holistic nature and individual variation of the healing experience. These findings suggest that many linear models used in researching touch therapies are not complex enough to capture the experience of participants. PMID- 11890386 TI - Complementary and alternative medicine use among colorectal cancer patients in Alberta, Canada. AB - CONTEXT: No population-based data are available on the use of complementary and alternative medicine (CAM) specifically among colorectal cancer patients. OBJECTIVE: To examine the prevalence and determinants of CAM use among colorectal cancer patients in Alberta, Canada. DESIGN: Population-based questionnaire. SETTING AND PARTICIPANTS: Patients (871 of 1240 surveyed), or their close relatives or friends, who were diagnosed with colorectal cancer in 1993 or 1995 in Alberta, Canada. MAIN OUTCOME MEASURES: Demographics, lifestyle, health status, symptoms and coping mechanisms, and attitudes about cancer cause, conventional treatments and practitioners, and CAM and practitioners. RESULTS: Seventy percent (871) of 1240 participants completed the questionnaire, and 49% used CAM. The most frequently used CAM therapies among users were psychological and spiritual therapies (65%), vitamins and minerals (46%), and herbs (42%). Sixty-eight percent of CAM users informed their medical doctors, and 69% used CAM after conventional care. Logistic regression suggested the strongest predictors of CAM use to be vegetarian diet, aged less than 50 years, female, having therapy options other than conventional treatment recommended by conventional doctors, experiencing changes in bowel habits orfatigue before diagnosis, and recommendation of chemotherapy. Nonsurviving patients were more likely to have used CAM than were survivors. CONCLUSIONS: Cancer patients are using CAM and communicating usage to physicians. This finding suggests that physicians should be prepared to discuss CAM with patients, and evidence-based information about CAM should be sought, including where CAM may pose risks. This study serves as a baseline for studies on the efficacy and safety of CAM. PMID- 11890387 TI - An integral approach to medicine. AB - In this article we suggest that despite decades of compelling research in suchfields as behavioral medicine and mind-body medicine, a more integral, less fragmented approach is still needed. We argue that one of the obstacles to realizing a more holistic-oriented medicine (ie, biopsychosociocultural) has been the lack of a comprehensive conceptual framework. We therefore propose the application in medicine of modern-day philosopher Ken Wilber's 4-quadrant model, which interfaces the dimensions of interior and exterior with those of the individual and collective. The article suggests that Wilber's framework offers a simple yet elegant heuristic tool for conceptualizing health and illness, investigating the efficacy of different treatment modalities, exploring the multifactorial nature of disease, and informing both research methodology and medical education. We further argue that this model has relevance for both the complementary and alternative as well as conventional medical fields, offering researchers, clinicians, and educators a way to clarify and operationalize otherwise vague concepts such as "holistic" and "integrative." PMID- 11890388 TI - Roger Walsh, MD, PhD essential spirituality for healing professionals. Interview by Bonnie Horrigan. PMID- 11890389 TI - Progress notes: University of Washington School of Medicine/Bastyr University. PMID- 11890390 TI - Maca: from traditional food crop to energy and libido stimulant. PMID- 11890391 TI - Growth of Holstein calves from birth to 90 days: the influence of dietary zinc and BLAD status. AB - The main objective of this study was to describe Holstein neonatal growth and development as influenced by dietary zinc supplementation and the CD18 genotype, both of which may affect immune competence. Holstein calves (n = 421), after being fed colostrum, were brought to a calf facility, randomly assigned to one of four zinc supplementation groups (control at 40 mg Zn/kg DM or the control diet supplemented with an additional 60 mg Zn/kg DM provided as either zinc sulfate, zinc lysine, or zinc methionine), weighed, and measured for morphometric growth parameters. Measurements were repeated at 30, 60, and 90 d. Calves were also genotyped for the presence of the mutant D128G CD18 allele, which, if present in two copies, causes bovine leukocyte adhesion deficiency. Zinc supplementation above 40 mg Zn/kg DM, regardless of the chemical form, did not accelerate growth (P > 0.25). Further, overall calf growth performance was not suppressed or improved (P > 0.4) in calves heterozygous at the CD18 locus relative to calves homozygous for the normal CD18 allele, although genotype negatively affected some morphometric measurements (P < 0.05). Using these data, quadratic models of early growth were generated as a preliminary step to develop growth criteria that will allow producers, veterinarians, and animal scientists to identify poor growth performance early in neonatal life. Such criteria provide the basis for tools to improve economic performance. PMID- 11890392 TI - Impacts on conceptus survival in a commercial swine herd. AB - An estimated 30 to 40% of potential piglets are lost before farrowing in U.S. or European pig breeds. Because these studies were conducted in limited numbers of university research herds, we decided to characterize the timing, pattern, and extent of conceptus loss in a commercial swine herd in Iowa (Pig Improvement Company; Camborough Line). Sows (parities 2 to 14) were slaughtered on d 25 (n = 83), 36 (n = 78), or 44 (n = 83) of gestation. These days coincide with periods before, during, and after uterine capacity becomes limiting to conceptus survival. At slaughter, numbers of corpora lutea (CL) and uterine horn length were determined, and conceptuses were removed and evaluated. Uterine horn length and CL number did not differ among these days of gestation, averaging 217 cm and 26.6 CL, respectively. In contrast, numbers of conceptuses decreased (P < 0.05) from 15.8 on d 25 to 12.9 on d 36, then remained relatively constant through d 44 (12.1). Thus, conceptus survival averaged 60.2% on d 25, 50.1% on d 36, and 46.3% on d 44, based on numbers of CL present. There was a positive correlation (P < 0.001; r = 0.50) between numbers of viable conceptuses on d 25 and ovulation rate, but this association was completely lost by d 36 (P > 0.10) when uterine capacity becomes limiting. In agreement with this premise, uterine horn length and conceptus number were not associated on d 25 but exhibited positive correlations (P < 0.05) on d 36 (r = 0.36) and d 44 (r = 0.40). On all 3 d examined, the numbers of viable conceptuses were not associated with fetal weight but were negatively correlated (P < 0.05) with placental weight. Compared with the commonly reported values for ovulation rate and percentage conceptus loss in university research herds, values from these production animals were extremely high. Data suggest that throughout this period, larger litters were associated with conceptuses exhibiting small placentae. These data lend support to the concept that increased placental efficiency (fetal weight/placental weight) may contribute to increased litter size in the pig. PMID- 11890394 TI - Effect of group size and feeder type on growth performance and feeding patterns in finishing pigs. AB - The effects of four group sizes (2, 4, 8, and 12 pigs per pen) and two single space feeder types (conventional and electronic feed intake recording equipment [FIRE]) on feed intake, growth performance, and feeding patterns were determined in 208 crossbred finishing pigs (equal numbers of barrows and gilts) between 84.4 (SD = 0.81) to 112.8 (SD = 1.08) kg BW over a 4-wk period. Pigs were given ad libitum access to a corn-soybean meal-based diet (15.9% CP; 0.79% lysine; 3,328 kcal ME/kg). The floor space allowance was 0.9 m2/pig for all treatments. Growth rates were not different for the two feeder types; however, feed intake was lower and gain:feed ratio higher for pigs on the FIRE feeders (P < 0.01). Feed intake, growth rate, and gain:feed ratio were not different (P > 0.05) among the group sizes. Number of feeder visits per day decreased and feed intake per visit, feeder occupation time per visit, feed consumption rate, and percentage of time the feeder was occupied increased with group size (P < 0.05). Feed intake per visit had the strongest correlation with daily feed intake (r = 0.54; P < 0.01) and was negatively correlated with gain:feed ratio (r = -0.38; P < 0.01). However, the correlations between growth performance and other feeding pattern traits were relatively weak (r < or = 0.30). As group size increased, diurnal variation in number of feeder visits and feed consumed per hour decreased. There was no difference in time spent sitting and standing between the two feeder types. The proportion of time spent eating was generally lower for the larger groups on both feeders. The proportion of time spent lying was similar across group sizes for pigs on the conventional feeders but was greater for pigs in the larger groups on the FIRE feeders. This study suggests that finishing pigs can maintain feed intake and growth rate by changing feeding behavior as group size increases from 2 to 12. PMID- 11890393 TI - An evaluation of the USDA standards for feeder cattle frame size and muscle thickness. AB - This study was conducted to determine the live weights at which large-, medium-, and small-framed feeder steers and heifers attain a degree of finish associated with a carcass quality grade of low Choice and to examine the relationship of feeder cattle muscle thickness to carcass yield grade traits. Feeder steers (n = 401) and heifers (n = 463) representing three age classes (calf, yearling, long yearling) were selected randomly at a commercial feedlot to exhibit wide ranges in frame size and muscularity. Individual weights were recorded and a panel of five experienced evaluators scored each animal for frame size, muscle thickness, and flesh condition. The cattle were finished on a high-concentrate finishing diet and harvested at an estimated carcass fat thickness of 10 mm. Final weights and USDA carcass grade data were collected for all cattle. Frame size scores effectively predicted finished weight at a marbling end point of Small(00) for both heifers (r2 = 0.89, SE = 16 kg) and steers (r2 = 0.94, SE = 13 kg). For heifers, the Small/Medium and Medium/ Large frame score intersects corresponded to live weights of 460 kg and 520 kg, respectively. For steers, the Small/Medium and Medium/Large frame score lines corresponded to live weights of 504 kg and 577 kg, respectively. These weights were greater than weights specified in the 1979 USDA grade standards. Evaluations of feeder cattle muscling, based on 1979 USDA Standards, were associated (P < 0.05) with differences in longissimus muscle area but were not related (P = 0.08) to differences in numerical carcass yield grades. An alternative muscle thickness classification scheme, involving the use of four thickness classes, was effective for stratifying feeder cattle according to eventual differences (P = 0.004) in carcass yield grade. Our findings suggest that USDA feeder cattle grade standards developed in 1979 are no longer adequate for describing today's population of feeder cattle. PMID- 11890395 TI - Ontogenic development of brown adipose tissue in Angus and Brahman fetal calves. AB - Brahman calves experience greater neonatal mortality than Angus calves if cold stressed. To establish a developmental basis for this, three fetuses of each breed type were taken at 96, 48, 24, 14, and 6 d before expected parturition, and at parturition. Overall fetal BW tended (P = 0.08) to be greater for Angus than for Brahman fetuses. There was no difference between breed types in total brown adipose tissue (BAT) mass or grams of BAT/kg BW. Brown adipocyte density decreased 56%, whereas lipogenesis from acetate and glucose in vitro decreased 97% during the last 96 d of gestation in both breed types. Glycerolipid synthesis from palmitate declined by 85% during the last trimester but still contributed 98% to total lipid synthesis at birth. The fetal age x breed interaction was significant for lipogenesis from glucose (P = 0.05) and palmitate (P = 0.005); rates were higher at 96 d before birth in Brahman BAT but declined to similar rates by birth. Uncoupling protein-1 (UCP1) mRNA tripled during gestation in both breed types (P = 0.002), whereas mitochondrial cross-sectional area did not change (P = 0.14) during gestation. Neither the breed nor the age x breed effect was significant (P > or = 0.24) for UCP1 mRNA concentration or mitochondrial cross-sectional area. In both breed types, a marked decrease in BAT UCP1 mRNA between 24 and 14 d prepartum was associated with a similar reduction in lipogenesis from palmitate and a noticeable change in BAT mitochondrial morphology, as the mitochondria became more elongated and the cristae became more elaborate. Uncoupling protein-1 mRNA initially was elevated in Angus tailhead s.c. adipose tissue, but was barely detectable by birth, and tended to be greater overall (P = 0.09) in Angus than in Brahman BAT. If uncoupling protein activity in s.c. adipose tissue persists after birth, then s.c. adipose tissue may contribute more to thermogenesis in Angus newborn calves than in Brahman calves. In contrast, we did not observe differences in ontogenic development of perirenal BAT that could explain the documented differences in thermogenic capacity between Angus and Brahman newborn calves. PMID- 11890396 TI - Uncoupling proteins and energy expenditure in mice divergently selected for heat loss. AB - The objective of this study was to determine whether variation in energy expenditure created by selection on heat loss is mediated by uncoupling protein-1 (UCP1) in brown adipose tissue. Divergent selection for heat loss developed lines of mice with high (MH) and low (ML) maintenance energy expenditure. Concentration of UCP1 mRNA in brown adipose tissue (BAT) was 93% greater in ML than in MH mice (P < 0.02). Two new lines of mice, KH and KL, were bred by backcrossing a UCP1 knockout gene into the MH and ML lines, respectively; KH and KL with both knock out (-/-) and wild type (+/+) UCP1 genotypes were generated. At 13 wk of age, KH mice exhibited greater heat loss (166 kcal x kg(0.75) x d(-1)) than KL mice (126.4 kcalkg(0.75) x d(-1)) regardless of the UCP1 knockout (P < 0.0001). Concentration of UCP2 mRNA in BAT was 74% greater in UCP1 knockout mice (-/-) than in wild type (+/+; P = 0.0001). We conclude that response to selection for increased energy expenditure was not mediated by increased expression or function of UCP1. PMID- 11890397 TI - Feeding lambs high-oleate or high-linoleate safflower seeds differentially influences carcass fatty acid composition. AB - Our objective was to determine effects of dietary high-oleate (Oleate; 76% 18:1) or high-linoleate (Linoleate; 78% 18:2) safflower seeds on fatty acids in muscle and adipose tissue of feedlot lambs. White-faced ewe lambs (n = 36) were fed a beet pulp, oat hay, and soybean meal basal diet (Control), blocked by BW, and allotted randomly to dietary treatments. Cracked safflower seeds were used in isocaloric and isonitrogenous replacement of beet pulp, oat hay, and soybean meal so that Oleate and Linoleate diets contained 5.0% additional fat. Fatty acids were determined in semitendinosus, longissimus dorsi (longissimus), and adipose tissue from the tail head (tailhead adipose tissue), adjacent to the 12th rib (s.c. adipose tissue), and kidney and pelvic fat (KPH adipose tissue) depots. Fatty acid data were analyzed within muscle and adipose tissue as a split-block design. Single degree of freedom orthogonal contrasts were used to compare treatment effects. Average daily gain, feed efficiency, and carcass characteristics did not differ (P = 0.15 to 0.96) across dietary treatments. Adipose tissue saturated fatty acids were greater (P = 0.04) for Controls but were not different (P = 0.36) in muscle. Trans-vaccenic acid (18:1(trans-11)) increased (P < 0.0001) with safflower supplementation and was greater (P < 0.0001) in Linoleate than in Oleate for both tissue types. Linoleate lamb had greater (P < 0.0001) PUFA than Oleate lamb in muscle and adipose tissue. Conjugated linoleic acids (CLA; cis-9, trans-11 and trans-10, cis-12) were greater (P < 0.0001) in muscle and adipose tissue of lambs fed safflower seeds. Lambs fed Linoleate had greater (P < 0.0001) CLA in adipose tissue and muscle than lambs fed Oleate. Saturated fatty acids were greater (P < 0.0001) in s.c. adipose tissue than in tailhead adipose tissue. Mono- and polyunsaturated fatty acids were greater (P < 0.0001) in tailhead adipose tissue than in s.c. adipose tissue. Weight percentages of 18:1(trans-11) ranked tailhead adipose tissue = KPH adipose tissue > s.c. adipose tissue and semitendinosus > longissimus, whereas CLA ranked tailhead adipose tissue > s.c. adipose tissue > KPH adipose tissue and semitendinosus > longissimus. Feeding mono- and polyunsaturated fatty acids increased tissue 18:1(trans-11) and CLA, which is a favorable change in regard to current human dietary guidelines. PMID- 11890398 TI - Effects of dietary lipid type on muscle fatty acid composition, carcass leanness, and meat toughness in lambs. AB - Isonitrogenous amounts of two protein sources differing in rumen degradation rate and in lipid composition were fed to sheep with or without a rapidly fermentable cereal grain. The effects on intake, carcass leanness, and muscle fatty acid (FA) composition were examined. Thirty-eight crossbred wether lambs (9 mo, 35 to 48 kg) were allocated by stratified randomization to six treatment groups: 1) basal diet of alfalfa hay:oat hay (20:80) ad libitum = basal; 2) basal + lupin (358 g DM/d) = lupin; 3) basal + fish meal (168 g DM/d) = fish meal; 4) basal + barley (358 g DM/d) = barley; 5) basal + barley + lupin (179 + 179 g DM/d) = barley/lupin; or 6) basal + barley + fish meal (179 + 84 g DM/d) = barley/ fish meal. Lambs were fed individually. Dietary treatments were imposed for 8 wk, and the supplements were offered at 2-d intervals. Daily feed intake and weekly BW of lambs were recorded. At the end of the feeding period lambs were slaughtered after an overnight fast. Hot carcass weight (HCW) and fat depth (GR; total fat and muscle tissue depth at 12th rib, 110 mm from midline) were recorded. At 24 h postmortem samples of longissimus thoracis (LT) and longissimus lumborum (LL) muscles were taken from chilled (4 degrees C) carcasses for the assessment of FA composition and meat tenderness, respectively. Lambs fed lupin or fish meal with or without barley had heavier slaughter weights (P < 0.004) and HCW (P < 0.001) than lambs fed basal or barley when initial BW was included as a covariate. The lupin diet also resulted in heavier carcasses (P < 0.05) than the fish meal or barley/fish meal diets. With GR as an indicator, fish meal and barley/ fish meal diets produced leaner carcasses (P < 0.01) than lupin and barley/lupin lambs. Long-chain n-3 FA content [20:5n-3 (P < 0.001), 22:5n-3 (P < 0.003), and 22:6n-3 (P < 0.001)] in the LT muscle were substantially higher with the fish meal and barley/fish meal diets, whereas muscle total n-6 FA was increased (P < 0.003) by lupin and barley/lupin compared with all other diets. Thus, increased muscle long chain n-3 FA content occurred without an increase in fatness measured as GR, whereas increased muscle n-6 FA content was associated with an increase in carcass fatness. Under these circumstances, a reduction in carcass fatness had no effect on meat tenderness measured as Warner-Bratzler shear force. PMID- 11890399 TI - Duration of feeding conjugated linoleic acid influences growth performance, carcass traits, and meat quality of finishing barrows. AB - Conjugated linoleic acid (CLA) was fed to growing-finishing barrows (n = 92) at 0.75% of the diet. A commercial CLA preparation (CLA 60) containing 60% CLA isomers was included at 1.25% to provide 0.75% CLA in the diet. The inclusion of CLA in diets was initiated at various BW and fed until slaughter. Growth, carcass, meat quality, physical, chemical, and sensory data were collected and analyzed. Treatments T1, T2, T3, and T4 included the last 0, 29, 56 and 87 kg, respectively, of weight gain before slaughter. Average daily gain and feed intake were not affected (P > or = 0.06) by CLA, but gain:feed responded quadratically (P = 0.05), over the entire BW gain (28 to 115 kg) with pigs of T2 and T3 having the greatest gain:feed. Loin muscle area increased (P = 0.01) linearly with increasing weight gain while fed CLA, and 10th rib, first rib, and last rib fat depth decreased (P < or = 0.05) linearly. Subjective quality measures on loin muscles increased linearly for marbling (P < 0.05) and tended to increase for firmness (P = 0.07) with increasing weight gain while barrows were fed CLA. Objective Hunter color values for loin chops from T1 and T4 were not different for L* (P = 0.12) or a* (P = 0.08) values but were higher (P < 0.05) for b* values with CLA feeding. Lipid oxidation values of loin muscle tissue were lower (P < 0.05) for pigs fed CLA (T1 vs T4). Increasing the period of weight gain while feeding CLA linearly increased (P < 0.01) saturated fatty acids and CLA isomers in loin tissue and linearly increased (P < 0.01) saturated fatty acids and CLA isomers in subcutaneous adipose tissue. Sensory panel characteristics of loin chops were not changed (P > 0.05) by feeding CLA. Increased gain:feed, increased loin muscle area, decreased fat depth, and improvements in marbling and firmness with CLA feeding could result in improved profitability of pork production systems. PMID- 11890400 TI - Growth performance, diet apparent digestibility, and plasma metabolite concentrations of barrows fed corn-soybean meal diets or low-protein, amino acid supplemented diets at different feeding level. AB - Two experiments, each with 36 barrows with high-lean-gain potential, were conducted to evaluate apparent nutrient digestibilities and performance and plasma metabolites of pigs fed corn-soybean meal diets (CONTROL) and low-protein diets. The low-protein diets were supplemented with crystalline lysine, threonine, tryptophan, and methionine either on an ideal protein basis (IDEAL) or in a pattern similar to that of the control diet (AACON). Amino acids were added on a true ileally digestible basis. The initial and final BW were, respectively, 31.5 and 82.3 kg in Exp. 1 and 32.7 and 57.1 kg in Exp. 2. In Exp. 1, the CONTROL and IDEAL diets were offered on an ad libitum basis or by feeding 90 or 80% of ad libitum intake. Pigs were fed for 55 d. In Exp. 2, the CONTROL, IDEAL, and AACON diets were offered on an ad libitum basis or by feeding 80% of the ad libitum intake. Pigs were fed for 27 d. Pigs fed the CONTROL diet had greater (P < 0.05) ADG and feed efficiency (G/F) than pigs fed the IDEAL (Exp. 1 and 2) and AACON diets (Exp. 2). As the level of feed intake decreased, ADG decreased (P < 0.05), but G/F tended to improve (P < 0.10) for pigs fed 90% of ad libitum in Exp. 1 and for pigs fed 80% of ad libitum in Exp. 2. In Exp. 1, the apparent total tract digestibilities of DM and energy were greater (P < 0.01) for pigs fed the IDEAL diet than for pigs fed the CONTROL diet. In Exp. 2, the apparent total tract digestibility of protein was greatest in pigs fed the CONTROL diet (P < 0.05) and was greater (P < 0.05) in pigs fed the AACON diet than in pigs fed the IDEAL diet. Plasma urea concentrations were lower in pigs fed the IDEAL diet than in pigs fed the CONTROL diet, regardless of feeding level. For pigs fed the CONTROL diet, plasma urea concentrations were lower when feed intake was 80% of ad libitum (diet level, P < 0.01). In summary, pigs fed the IDEAL and the AACON diets gained less and had lower plasma urea concentrations than pigs fed the CONTROL diet. Based on these data, it seems that the growth potential of pigs fed the IDEAL and AACON diets may have been limited by a deficiency of lysine, threonine, and(or) tryptophan and that the amino acid pattern(s) used was not ideal for these pigs. PMID- 11890401 TI - Body composition and tissue accretion rates of barrows fed corn-soybean meal diets or low-protein, amino acid-supplemented diets at different feeding levels. AB - Two experiments, each with 39 high-lean-gain potential barrows, were conducted to evaluate the organ weights, body chemical composition, and tissue accretion rates of pigs fed corn-soybean meal diets (CONTROL) and low-protein diets supplemented with crystalline lysine, threonine, tryptophan, and methionine either on an ideal protein basis (IDEAL) or in a pattern similar to that of the control diet (AACON). Amino acids were added on a true ileally digestible basis. The initial and final BW were, respectively, 31.5 and 82.3 kg in Exp. 1 and 32.7 and 57.1 kg in Exp. 2, and pigs were fed for 55 and 27 d in Exp. 1 and 2, respectively. In Exp. 1, the CONTROL and IDEAL diets were offered on an ad libitum basis, or by feeding 90 or 80% of ad libitum intake. In Exp. 2, the CONTROL, IDEAL, and AACON diets were offered on an ad libitum basis, or by feeding 80% of the ad libitum intake. Three pigs were killed at the start of the experiments and three from each treatment were killed at the end of each experiment to determine body chemical composition. In both trials, the whole-body protein concentration (g/kg) and the accretion rates of protein (g/d) were greater (P < 0.05) for pigs fed the CONTROL than for pigs fed the IDEAL and AACON diets. In Exp. 1, pigs fed the CONTROL diet had a trend (P < 0.10) for greater water and lower lipid concentration and had greater (P < 0.05) water and ash accretion rates. Whole body protein concentration was greatest (P < 0.05) in pigs fed at 80% of ad libitum, but protein, water, and ash accretion rates were greatest (P < 0.05) in pigs allowed ad libitum access to feed. In summary, pigs fed the IDEAL and the AACON diets had less protein in the body and lower protein accretion rates than pigs fed the CONTROL diet. It seems that reductions in protein deposition in pigs fed the IDEAL and AACON diets may have been due to a deficiency of one or more essential amino acids or possibly to increases in the NE for metabolic processes leading to increases in adipose tissue deposition. PMID- 11890402 TI - Effect of dietary levels of vitamin E (all-rac-tocopheryl acetate) with or without added fat on weanling pig performance and tissue alpha-tocopherol concentration. AB - Two experiments involving 496 cross-bred pigs evaluated the efficacy of various dietary levels of vitamin E, with or without supplemental fat, on postweaning pig performance and weekly serum and terminal tissue alpha-tocopherol concentrations. The first trial involved 248 pigs weaned at an average of 15 d of age and 4.8 kg BW. The experiment was a randomized complete block design conducted in seven replicates. Vitamin E was added as dl-alpha-tocopheryl acetate at 0, 20, 40, 60, 80, 100, 150, or 200 IU/kg diet. Pigs were bled initially and at 7-d intervals for a 42-d period. Liver and s.c. adipose tissue samples were collected from six pigs per treatment group at 42 d. In Exp. 2, a 2 x 4 factorial arrangement of treatments in a randomized complete block design was conducted in seven replicates. The experiment used a total of 248 pigs weaned at 19 d of age and averaged 6.4 kg BW. Four vitamin E levels (0, 20, 40, and 60 IU/kg diet) and two added fat levels of 0 or 5% were fed for 35 d. Four pigs per treatment pen were bled weekly, and at 35 d a total of four pigs per treatment group were killed and liver, heart, and s.c. adipose tissues were collected and analyzed for alpha tocopherol. The basal diet in both experiments contained an average 7.9 IU for period 1, and later diets averaged 11.0 IU vitamin E/kg. In both experiments serum alpha-tocopherol concentrations declined from weaning to 7 d after weaning and continued to decline each week after weaning when the basal diets were fed. Serum alpha-tocopherol concentrations increased (P < 0.01) each week as the dietary vitamin E level increased in both experiments. In Exp. 2, when fat was added to the diet serum alpha-tocopherol concentrations were higher (P < 0.01) than in diets without added fat. Liver, heart muscle, and adipose tissue alpha tocopherol concentrations increased (P < 0.01) as vitamin E level increased, but at the higher dietary vitamin E level the liver surpassed the adipose tissue in its alpha-tocopherol concentration. Liver and adipose alpha-tocopherol concentrations were higher (P < 0.01) when fat was added to the diet. These results indicate that supplementation of 40 to 60 IU/kg diet with added fat resulted in a relatively constant balance of serum and tissue concentration of alpha-tocopherol during the nursery period, but when fat was not supplemented a dietary vitamin E level of 80 to 100 IU/kg diet may be needed. The current NRC recommendations for vitamin E for the pig from 5 to 20 kg BW may need to be reevaluated. PMID- 11890403 TI - Tributyrin and lactitol synergistically enhanced the trophic status of the intestinal mucosa and reduced histamine levels in the gut of nursery pigs. AB - This study determined whether tributyrin and lactitol could synergistically facilitate the transition from milk to solid feed in nursery pigs. At 21 d after birth, 64 piglets were moved from the piggery to a production barn and fed a medicated diet. At 28 d after birth, the piglets were weighed and allotted into four groups and fed a standard nonmedicated diet (control) or the control diet with tributyrin (butanoic acid 1,2,3-propanetriyl ester; 10 g/kg), or with lactitol (beta-D-galactopyranosyl-(1-->4)-D-sorbitol; 3 g/kg), or with tributyrin (10 g/kg) plus lactitol (3 g/kg). On d 0, 14, and 42 after being fed the control or experimental diets, the animals were weighed, and animal health, feed intake, and feed conversion ratio were determined. On d 42, four piglets from each treatment were killed to measure the empty and full weight of the gut, as well as the weights of the liver and kidneys. The jejunum and cecum were sampled to analyze the luminal concentrations of lactic acid; short-chain fatty acids; and mono-, di-, and polyamines and to assess the mucosal status. Mortality after 42 d ranged from 19% for animals fed the control diet, to 6% for animals fed the tributyrin or lactitol diets, and to 0% for animals fed the tributyrin+lactitol diet. After 14 d, the ADG was 127% greater (P < 0.05) in animals fed the tributyrin+lactitol diet than in animals fed the control or tributyrin diets. After 42 d, animals fed the tributyrin+lactitol diet were heavier (P < 0.05) than animals fed the tributyrin diet. At slaughter, no differences (P > 0.05) in organ weights were observed. With the exception of animals fed the lactitol diet, wherein cecal lactic acid levels increased threefold (P < 0.01), the luminal concentrations of lactic acid and short-chain fatty acids were not different (P > 0.05). Among the various amines analyzed, the only response (P < 0.05) was a 66 and 49% decrease in histamine levels in the jejunum and cecum, respectively, in animals fed the tributyrin+lactitol diet compared to the control diet. In the jejunum of animals fed the lactitol or tributyrin+lactitol diets, the length of the villi was increased by 12% (P < 0.05) compared to animals fed the control diet, whereas the tributyrin diet did not have any effect on the villi (P > 0.05). In the cecum, the depths of the crypts were reduced (P < 0.001) by 18% in animals fed the lactitol diet and 45% in animals fed the tributyrin or tributyrin+lactitol diets compared to animals fed the control diet. In conclusion, a diet containing tributyrin and lactitol as nutribiotics resulted in lower histamine levels in the jejunum and cecum, as well as longer jejunal villi and shallower cecal crypts. PMID- 11890404 TI - Effect of high temperature and low-protein diets on the performance of growing finishing pigs. AB - The effects of reducing CP level in combination with an increase in ambient temperature (29 vs 22 degrees C) on performance and carcass composition were studied in a factorial arrangement of treatments involving 66 Pietrain x (Landrace x Large White) barrows from 27 to 100 kg BW. Animals were fed at each temperature one of three experimental diets that provided 0.85 or 0.70 g of digestible lysine per megajoule of NE, in the growing (27 to 65 kg) and the finishing (65 to 100 kg) phases, respectively. Diet 1 was a corn, wheat, and soybean meal diet formulated without crystalline AA; CP levels were 20.3 and 17.6% for the growing and the finishing phases, respectively. In Diets 2 and 3, CP level was reduced by substituting part of the soybean meal with corn and wheat (Diet 2), or with corn, wheat, and 4% fat (Diet 3). Diets 2 and 3 were supplemented with AA and balanced according to the ideal protein concept. The CP levels of Diets 2 and 3 were, respectively, 15.8 and 16.3% in the growing phase, and 13.4 and 13.8% in the finishing phase. Pigs were housed individually and had free access to feed and water. The ADFI was measured daily, and animals were weighed weekly. Carcass composition was measured at slaughter (100 kg BW). Increasing ambient temperature from 22 to 29 degrees C resulted in a 15% reduction in ADFI and 13% lower ADG. Leaner carcasses (P < 0.01) were obtained at 29 degrees C (22.8 vs 24.8% carcass fat). At 22 degrees C, ADFI was lower (P < 0.05) for the low-CP diets, but daily NE intake, ADG, and carcass composition were not affected (P > 0.05). At 29 degrees C, ADFI was not different (P > 0.05) between diets and daily NE intake was higher (P < 0.05) with Diet 3 than with Diet 1, and the difference was more important during the finishing period than during the growing period. Using the model ADFI = a BWb, estimates of b were 0.65, 0.53, and 0.53 at 22 degrees C and 0.50, 0.44, and 0.50 at 29 degrees C, for Diets 1, 2, and 3, respectively. The higher NE intake for Diet 3 at 29 degrees C did not improve ADG (P > 0.05) but increased mainly fat deposition. These results indicate that a 4 percentage unit reduction of dietary CP level reduces N excretion (minus 37%) but does not affect growth and carcass composition as long as the ratio between essential AA and NE are kept optimal. In addition, diets with reduced CP limit the effect of high ambient temperature on ADFI. Finally, our results demonstrate the significance of using NE, rather than DE or ME, for formulating diets. PMID- 11890405 TI - Relative bioactivity of dietary RRR- and all-rac-alpha-tocopheryl acetates in swine assessed with deuterium-labeled vitamin E. AB - This study evaluated the relative bioactivities of natural and synthetic stereoisomers of alpha-tocopherol in swine. Deuterium-labeled vitamin E (150 mg each of d3-RRR- [natural] and d6-all-rac- [synthetic] alpha-tocopheryl acetates) was administered orally to adult female pigs (n = 3) with the morning feed. Blood samples were obtained at 0, 3, 6, 9, 12, 36, 48, and 72 h after the dose. The time of maximum plasma d3-alpha-tocopherol concentration (0.486 microg/mL) occurred at 12 h, and d6-alpha-tocopherol peaked earlier (at 9 h) and at a lower (P < 0.05) concentration (0.288 microg/mL). The d3-/d6-alpha-tocopherol ratio increased from 1.35 (SD = 0.73) at 3 h after dosing to 2.0 (SD = 0.14) at 72 h (P = 0.03). The plasma disappearance rates of d3- and d6-alpha-tocopherols (post maximum concentrations) were similar and were estimated to be 0.013 microg/mL per hour. In summary, swine discriminated between RRR- and all-rac-alpha-tocopherols, which resulted in an approximately twofold higher plasma alpha-tocopherol concentration arising from the RRR-form. This 2:1 ratio of RRR- to all-rac- is higher than the currently accepted USP definition of RRR-:all-rac- of 1.36:1.00. PMID- 11890407 TI - Relationship between serum insulin-like growth factor-I and genotype during the postpartum interval in beef cows. AB - The objective of this study was to determine the effect of genotype and week postpartum on serum concentrations of IGF-I, body condition score (BCS), BW, and ovarian function in beef cows. Cows from the following genotypes were utilized in two consecutive years: Angus (A x A; n = 9), Brahman (B x B; n = 10), Charolais (C x C; n = 12), Angus x Brahman (A x B; n = 22), Brahman x Charolais (B x C; n = 19) and Angus x Charolais (A x C; n = 24). Serum concentrations of IGF-I, BCS, and BW were determined between wk 2 and 9 postpartum. Rectal ultrasound was used to determine days postpartum to first medium (6 to 9 mm) and first large (> or = 10 mm) follicle. Averaged across genotype, BCS decreased (P < 0.05) from 5.0 +/- 0.1 on wk 3 to 4.8 +/- 0.1 on wk 6 postpartum, and BW decreased (P < 0.05) between wk 2 and 3 and again between wk 4 and 9 postpartum. Averaged over year and week postpartum, serum IGF-I concentrations were greatest (P < 0.05) in B x B cows (46 +/- 5 ng/mL) compared with all other genotypes; lowest in A x A (12 +/- 4 ng/mL), C x C (13 +/- 4 ng/mL), and A x C cows (18 +/- 3 ng/mL); and intermediate (P < 0.05) in A x B (28 +/- 3 ng/mL) and B x C (26 +/- 3 ng/mL) cows compared with all other genotypes. Serum IGF-I concentrations did not change (P > 0.10) with week postpartum in C x C, A x A, and A x C cows, but increased (P < 0.05) between wk 2 and 7 postpartum in B x C, A x B, and B x B cows. Average interval to first medium (16 +/- 2 d) and first large (35 +/- 2 d) follicle did not differ (P > 0.10) among genotypes. Serum IGF-I concentrations correlated with BCS (r = 0.53 to 0.72, P < 0.001) but not with days to first large follicle (r = 0.19 to -0.22, P > 0.10). Averaged across genotypes, cows that lost BCS postpartum had lower (P < 0.01) serum IGF-I concentrations. Cows that calved with adequate BCS (i.e., > or = 5) had greater (P < 0.01) serum IGF-I concentrations postpartum than cows that calved with inadequate BCS (i.e., < 5) but days to first large and medium follicle did not differ (P > 0.10). In conclusion, concentrations of IGF-I in serum differed among genotypes and were associated with BCS but not days to first large or medium follicle in postpartum beef cows. PMID- 11890406 TI - Soybean meal from roundup ready or conventional soybeans in diets for growing finishing swine. AB - Dehulled soybean meal prepared from genetically modified, herbicide (glyphosate) tolerant Roundup Ready soybeans containing the CP4 EPSPS protein and near isogenic conventional soybeans were assessed in an experiment with growing finishing pigs. The soybeans were grown in the yr 2000 under similar agronomic conditions except that the Roundup Ready soybeans were sprayed with Roundup herbicide. Both were processed at the same plant. The composition of the two types of soybeans and the processed soybean meal were similar. Corn-soybean meal diets containing conventional or Roundup Ready soybean meal and fortified with minerals and vitamins were fed to 100 cross-bred pigs from 24 to 111 kg BW. Diets contained approximately 0.95% lysine initially and were reduced to 0.80 and 0.65% lysine when pigs reached 55 and 87 kg BW, respectively. There were 10 pens (five pens of barrows and five pens of gilts) per treatment with five pigs per pen. All pigs were scanned at 107 kg mean BW and all barrows were killed at the end of the test for carcass measurements and tissue collection. Rate and efficiency of weight gain, scanned backfat and longissimus area, and calculated carcass lean percentage were not different (P > 0.05) for pigs fed diets containing conventional or Roundup Ready soybean meal. Gilts gained slower, but they were more efficient and leaner (P < 0.05) than barrows. Responses to the type of soybean meal were similar for the two sexes with no evidence of a diet x sex interaction for any of the traits. In most instances, carcass traits of barrows were similar for the two types of soybean meal. Longissimus muscle samples from barrows fed conventional soybean meal tended (P = 0.06) to have less fat than those fed Roundup Ready soybean meal, but water, protein, and ash were similar. Sensory scores of cooked longissimus muscles were not influenced (P > 0.05) by diet. The results indicate that Roundup Ready soybean meal is essentially equivalent in composition and nutritional value to conventional soybean meal for growing-finishing pigs. PMID- 11890408 TI - Leptin expression in the ovine mammary gland: putative sequential involvement of adipose, epithelial, and myoepithelial cells during pregnancy and lactation. AB - We examined the ability of the ovine mammary gland to synthesize leptin throughout pregnancy and lactation. Leptin gene expression was assayed by real time reverse transcription and polymerase chain reaction in mammary gland from ewes at 15, 80, 106, 112, or 141 d of pregnancy and at 0 (30 min after parturition), 3, 48, or 70 d of lactation. Leptin mRNA level was high at the beginning (the first 80 d) and at the end of pregnancy and was lower at mid pregnancy and throughout lactation. Furthermore, during these periods of mammary leptin expression, the location of leptin protein, as determined by immunohistochemical analysis, changed within mammary tissue. It was located in adipose cells during early stages of pregnancy, in epithelial cells after full cell differentiation just before parturition, and in myoepithelial cells after parturition. These data, compared with published data on leptin receptor gene expression, provide evidence that leptin could be produced by different cell types of the mammary gland and could act as a paracrine factor on mammary cell growth and differentiation via adipose-epithelial cells and myoepithelial epithelial cell interactions. PMID- 11890409 TI - Interrelationships among conceptus size, uterine protein secretion, fetal erythropoiesis, and uterine capacity. AB - The interrelationships among d-11 conceptus size, d-105 placental weight, placental efficiency (the ability of the placenta to support fetal growth and development), fetal erythropoiesis, and uterine capacity were examined in 1/2 Meishan, 1/2 White crossbred gilts that were unilaterally ovariohysterectomized at 90 to 100 d of age. In Exp. 1, gilts were mated after at least one normal estrous cycle and then slaughtered at 105 d of gestation and number of fetuses and CL, placental weights, fetal weights, hematocrits, fetal plasma iron, and fetal plasma folate were measured. In Exp. 2, gilts were mated and plasma progesterone was measured on d 2 and 3 of gestation. On d 11, the length of the remaining uterine horn was recorded and the uterine horn was flushed with minimal essential medium. Number of CL, conceptus number, conceptus diameters, and total uterine flush retinol-binding protein (tRBP), acid phosphatase (tAP), and folate binding protein (tFBP) were measured. Gilts were mated again and slaughtered at 105 d of pregnancy and the same traits measured in Group 1 were recorded. Plasma progesterone concentrations on d 2 and 3 were correlated with average conceptus diameter on d 11 (r = 0.60, P < 0.01, for each day). In contrast, tRBP (r = 0.49, P < 0.01), tAP (r = 0.53, P < 0.01), and tFBP (r = 0.51, P < 0.01) in uterine flushings on d 11 were only correlated with d-3 plasma progesterone concentrations. No correlations between d-11 average conceptus diameter or d-11 uterine length with d-105 uterine capacity were observed. Uterine capacity was negatively correlated with placental weight, fetal weight and fetal hematocrit (r = -0.36, P < 0.01; r = -0.44, P < 0.01; r = -0.32, P < 0.01; respectively). Hematocrits were correlated with fetal plasma iron (r = 0.50, P < 0.01) and folates (r = 0.44, P < 0.01). Hematocrit, plasma iron, and plasma folate were each correlated with residual fetal weights after adjusting for placental weight (a measure of placental efficiency), and accounted for 11% of the variation in this trait. These data suggest that conceptus diameter and uterine protein secretion on d 11 may be influenced by the onset of progesterone secretion by the CL, but do not support an influence of conceptus growth during early pregnancy on uterine capacity. These results also suggest that reducing placental and fetal weights will likely result in increased uterine capacity. PMID- 11890410 TI - Leptin concentrations in periparturient ewes and their subsequent offspring. AB - Leptin is an adipocyte-derived hormone that suppresses feed intake and increases energy expenditure. Leptin is also involved in regulating body temperature. Thus, the presence of leptin in milk, which can be absorbed through the gut of neonates immediately after birth, may aid in the survival of neonates born in cold weather. Our objectives were to determine the temporal relationship between concentrations of leptin in postpartum ewe blood serum and ewe milk serum, and to determine whether ewe blood and milk serum leptin concentrations were correlated with concentrations of leptin in lamb blood serum in their off-spring. Approximately 1 wk before the expected date of lambing, blood samples, weights, and body condition scores (BCS; 0 to 5 scale) were collected from 27 mixed-parity ewes. Following parturition, ewe blood and milk samples were collected within 2 h of parturition (d 0), 12 h (d 0.5) and 24 h (d 1) after parturition, again on d 5, and weekly thereafter until d 47. Lambs were blood-sampled and weighed within 2 h of parturition (d 0), bled daily until d 5, and bled and weighed weekly thereafter to d 47. Prior to lambing, ewe blood serum leptin was positively correlated with congruent BCS (r2 = 0, 10, P = 0.06), but not weight (P = 0.14). Following parturition, ewe blood serum leptin was positively correlated with BCS, weight, and milk serum leptin (r2 = 0.14, P < 0.0001, r2 = 0.12, P < 0.0001, and r2 = 0.028, P = 0.04). Leptin in milk serum was correlated with ewe weight (r2=0.05, P = 0.007) but not ewe BCS (P = 0.7); however, concentrations of leptin in both ewe blood and milk serum varied with day of lactation (P = 0.0001), being maximal within 24 h of parturition and declining to nadir concentrations by d 5. Leptin in lamb serum was correlated with milk serum leptin, (r2 = -0.05; P = 0.001), but not ewe blood serum leptin (P = 0.5). Concentrations of leptin in lamb serum increased from birth to d 5 and declined thereafter to nadir concentrations by d 19. Elevated concentrations of leptin in milk during the early stages of lactation may provide a mechanism for thermoregulation, satiation, and homeostatic endocrine control in the neonate. PMID- 11890411 TI - Growth hormone response to a novel growth hormone-releasing tripeptide in horses: interaction with gonadotropin-releasing hormone, thyrotropin-releasing hormone, and sulpiride. AB - A series of experiments was performed to determine the factor(s) responsible for an apparent inhibition of GH secretion in mares administered the GH secretagogue EP51389 in combination with GnRH, thyrotropin-releasing hormone (TRH), and sulpiride. Experiment 1 tested the repeatability of the original observation: 10 mares received EP51389 at 10 microg/kg BW; five received TRH (10 microg/kg BW), GnRH (1 microg/kg BW), and sulpiride (100 microg/kg BW) immediately before EP51389, and five received saline. The mixture of TRH, GnRH, and sulpiride reduced (P = 0.0034) the GH response to EP51389, confirming the inhibitory effects. Experiment 2 tested the hypothesis that sulpiride, a dopamine antagonist, was the inhibitory agent. Twelve mares received EP51389 as in Exp. 1; six received sulpiride before EP51389 and six received saline. The GH responses in the two groups were similar (P > 0.1), indicating that sulpiride was not the inhibitory factor. Experiment 3 tested the effects of TRH and(or) GnRH in a 2 x 2 factorial arrangement of treatments. Three mares each received saline, TRH, GnRH, or the combination before EP51389 injection. There was a reduction (P < 0.0001) in GH response in mares receiving TRH, whereas GnRH had no effect (P > 0.1). Given those results, Exp. 4 was conducted to confirm that TRH was inhibitory in vivo as opposed to some unknown chemical interaction of the two compounds in the injection solution. Twenty mares received TRH or saline and(or) EP51389 or saline in a 2 x 2 factorial arrangement of treatments. Injections were given separately so that the two secretagogues never came in contact before injection. Again, TRH reduced (P < 0.0001) the GH response to EP51389. In addition, TRH and EP51389 each resulted in a temporary increase in cortisol concentrations. Experiment 5 tested whether TRH would alter the GH response to GHRH itself. Twelve mares received porcine GHRH at 0.4 microg/kg BW; six received TRH prior to GHRH and six received saline. After adjustment for pretreatment differences between groups, the GHRH-induced GH response was completely inhibited (P = 0.068) by TRH. Exp. 6 was a repeat of Exp. 5, except geldings were used (five per group). Again, pretreatment with TRH inhibited (P < 0.0001) the GH response to GHRH. In conclusion, TRH inhibits the GH response not only to EP51389 but also to GHRH in horses, and in addition to its known secretagogue action on prolactin and TSH it may also stimulate ACTH at the dosage used in these experiments. PMID- 11890412 TI - Human acylation-stimulating protein and lipid biosynthesis in bovine adipose tissue explants. AB - Human acylation-stimulating protein (hASP) up-regulates triacylglycerol synthesis in human adipocytes. The objectives of this research were 1) to determine the effect of hASP on triacylglycerol synthesis in bovine adipose explants and 2) to determine whether nutritional status influences the sensitivity of adipose tissue to hASP. Fresh s.c. adipose tissue was sectioned into 20- to 30-mg explants and incubated for 1 to 6 h in M199 media containing 3% BSA and either 0.75 mM [1 14C]palmitate, 0.75 mM [9, 10-3H]oleate, or 2.5 mM [1-14C] acetate, as well as hASP and(or) insulin. The explants were extracted, and lipid fractions were separated by TLC and quantified by liquid scintillation. Acetate incorporation into lipids increased 15 to 30%, and palmitate or oleate incorporation increased 10 to 25%, when explants were exposed to hASP, although this response was not significant in every experiment. Insulin increased triacylglycerol synthesis in some experiments, but not in others. Our interpretation is that acylation stimulating protein (ASP) can mildly enhance triacylglycerol synthesis in bovine adipose tissue. To fulfill the second objective, nine 9-mo-old steers were housed individually for two periods of 3 wk each. During the first period, four of the nine steers were fed to 50% of NEm requirement and the other five consumed the same diet ad libitum. After the first period, all steers consumed feed ad libitum for 2 wk and were assigned the opposite ration for the second period. Steers gained 40.5 kg BW when allowed ad libitum access to feed but lost 30.2 kg BW when feed intake was restricted (SE = 7.84; P < 0.01). At the end of each period, s.c. adipose tissue was sectioned into explants and incubated as described above. Four explants per steer per period were used to test effects of insulin (0 and 1 nM) and hASP (0, 0.01, 0.1, and 1 microM). Insulin did not influence incorporation of acetate or oleate. Acetate incorporation (P < 0.32) was 0.99, 1.03, 1.04, and 1.10 nmol x mg(-1) h(-1) (SE = 0.13) and oleate incorporation (P < 0.01) was 0.347, 0.357, 0.353, and 0.420 nmol x mg(-1)h(-1) (SE = 0.022) for 0, 0.01, 0.1, and 1 microM hASP, respectively. Feed restriction reduced (P < 0.01) acetate and oleate incorporation by 95 and 40%, respectively. No interactions among feed intake, insulin, and hASP were detected. In conclusion, the effect of hASP on fatty acid esterification is not influenced by feed restriction. PMID- 11890414 TI - Evaluation of hay-type and grazing-tolerant alfalfa cultivars in season-long or complementary rotational stocking systems for beef cows. AB - Alfalfa (Medicago sativa L.) persistence and forage and cow-calf production were evaluated on pastures containing smooth bromegrass with or without grazing tolerant or hay-type alfalfa cultivars rotationally stocked in either a season long or complementary system. In 1997, six 2.02-ha pastures were seeded with smooth bromegrass, a mixture of a grazing-tolerant alfalfa (Amerigraze variety) and smooth brome-grass, or a mixture of a hay-type alfalfa (Affinity variety) and smooth bromegrass to be used in season-long stocking systems. Four 2.02-ha pastures were seeded with smooth bromegrass on 1.21 ha of each pasture, and mixtures of either the grazing-tolerant or hay-type alfalfa cultivars and smooth bromegrass on the 0.81 ha of each pasture to be used in complementary stocking systems. All 10 pastures were divided into 10 paddocks and rotationally strip stocked at 1.98 cow-calf units/ha with crossbred cows and calves for 120 and 141 d starting May 18, 1998 (yr 1), and May 6, 1999 (yr 2), respectively. Each year, first harvest forage was harvested as hay from 40% of all 10 pastures, this being the portions of the pasture seeded with the alfalfa-smooth brome-grass mixtures for pastures with the complementary stocking systems. In yr 1 and 2, the remaining 60% of each pasture was grazed for the first 44 and 54 d, and 100% of each pasture was grazed on d 45 to 120 and d 55 to 141, respectively. Proportions of alfalfa in the live dry matter of pastures seeded with the grazing-tolerant and hay-type alfalfa cultivars decreased by 70 and 55% in paddocks stocked season long and by 60 and 42% in paddocks used for complementary stocking (alfalfa cultivar, P < 0.05; stocking system, P < 0.05) in yr 1, but decreased by 72% across cultivars and stocking systems in yr 2. Total (P < 0.08) forage masses in September of yr 1 and in August of yr 2 were greater in pastures in which alfalfa paddocks were stocked season-long than in those with complementary alfalfa stocking. Grazing of alfalfa in grass mixtures increased calf and total cow/calf weight gains in comparison with grazing of smooth bromegrass, but alfalfa persistence, measured as a proportion of the live dry matter, was not affected by alfalfa cultivar. PMID- 11890413 TI - Relationships of metabolic hormones and serum glucose to growth and reproductive development in performance-tested Angus, Brangus, and Brahman bulls. AB - Understanding mechanisms that regulate growth and reproduction are important for improving selection strategies in cattle. In this study, Angus, Brangus, and Brahman bulls (n = 7 per breed) of similar age were selected from a group of 65 weanlings. Bulls were evaluated after weaning (i.e., approximately 6 mo of age) for 112 d for serum concentrations of metabolic hormones and glucose, growth, and reproductive traits. Performance data and blood sera were collected on d 0, 28, 56, 84, and 112. Sera were also collected in periods from d 50 to 59 (56D) and 103 to 112 (112D). Angus bulls were heavier (P < 0.05) throughout the study than Brahman bulls and were heavier than Brangus bulls on d 56, 84, and 112. Initial and final BW for Angus, Brangus, and Brahman bulls were 292.7, 260.6, and 230.4 and 468.3, 435.6, and 350.7 +/- 12 kg, respectively. Conversely, Brahman bulls had greater hip height (P < 0.05) than Brangus, and Brangus were taller (P < 0.05) than Angus. Angus bulls had the greatest (P < 0.05) scrotal circumference (SC) and Brahman bulls the least. Mean SC across days was 31.5, 29.7, and 25.0 +/ 0.6 cm for the three respective breeds. Serum testosterone was greater (P < 0.01) in Angus and Brangus bulls (10.0 and 8.9 +/- 1.4 ng/mL) than in Brahman bulls (4.0 +/- 1.4 ng/mL) throughout the study. After d 112, 100, 86, and 57% of the Angus, Brangus, and Brahman bulls passed a breeding soundness exam (P = 0.51). Serum concentrations of IGF-I and leptin were greater (P < or = 0.06) in Angus bulls on d 56, 84, and 112 than in Brangus and Brahman bulls. Serum concentrations of GH (P < 0.08) and glucose (P < 0.03) were greater in Brangus bulls than in Angus or Brahman bulls throughout the study. Prediction analyses suggested that serum concentrations of leptin could be used to predict (P < or = 0.08) BW and SC (R2 > 0.82) in the 56D and 112D periods among these breeds. Leptin was also useful in predicting (P < or = 0.09) serum concentrations of GH and testosterone in the 112D period (R2 > 0.32). Residual correlation analyses with the effect of breed removed suggested that leptin was correlated (r > or => 0.53, P < 0.05) with both SC and serum testosterone. Angus and Brahman cattle differ in phenotype, level of adiposity, and rate of sexual development. Data herein suggest that these characteristics could be due to varying mechanisms by which metabolic hormones such as leptin, GH, and(or) IGF-I are regulated. PMID- 11890415 TI - Effects of supplementation on intake, digestion, and performance of beef cattle consuming fertilized, stockpiled bermudagrass forage. AB - Experiments were conducted to determine the effects of increasing supplement protein concentration on performance and forage intake of beef cows and forage utilization of steers consuming stockpiled bermudagrass forage. Bermudagrass pastures were fertilized with 56 kg of N/ha in late August. Grazing was initiated during early November and continued through the end of January each year. Treatments for the cow performance trials were: no supplement or daily equivalents of 0.2, 0.4, and 0.6 g of supplemental protein per kilogram of BW. Supplements were formulated to be isocaloric, fed at the equivalent of 0.91 kg/d, and prorated for 4 d/wk feeding. Varying the concentration of soybean hulls and soybean meal in the supplements created incremental increases in protein. During yr 1, supplemented cows lost less weight and condition compared to unsupplemented animals (P < 0.05). During yr 2, supplemented cows gained more weight (P = 0.06) and lost less condition (P < 0.05) compared to unsupplemented cows. Increasing supplement protein concentration had no affect on cumulative cow weight change or cumulative body condition score change. Forage intake tended to increase (P = 0.13, yr 1 and P = 0.07, yr 2) in supplemented cows. Supplement protein concentration did not alter forage intake. In a digestion trial, four crossbred steers were used in a Latin square design to determine the effects of supplement protein concentration on intake and digestibility of hay harvested from stockpiled bermudagrass pasture. Treatments were no supplement; or 0.23, 0.46, and 0.69 g of supplemental protein per kilogram of BW. Forage intake increased (P < 0.05) 16% and OM intake increased (P < 0.01) 30% in supplemented compared to unsupplemented steers. Diet OM digestibility increased (P = 0.08) 14.5% and total digestible OM intake increased (P < 0.05) 49% in supplemented compared to unsupplemented steers. Supplement protein concentration did not alter forage intake, total digestible OM intake, or apparent digestibility of OM or NDF. During the initial 30 d after first killing frost, beef cows did not respond to supplementation. However, later in the winter, supplementation improved utilization of stockpiled bermudagrass forage. PMID- 11890416 TI - A comparison of enzymatic and molecular approaches to characterize the cellulolytic microbial ecosystems of the rumen and the cecum. AB - We used RNA probes and enzyme activities to compare the cellulolytic microbial ecosystems of the rumen and the cecum. Four rumen- and cecum-cannulated wethers were fed a diet of barley plus hay (60:40). Digesta samples were collected 1 h before feeding and 3, 6, and 9 h after feeding for measurements on microbial populations, and 1 h before feeding and 3 and 6 h after feeding for digestion measurements, pH, and VFA. Polysaccharidase and glycosidase specific activities of solid-adherent microorganisms were measured respectively by the amount of reducing sugars released from xylan or avicel or p-nitrophenol from the p nitrophenol derivatives of xylose and glucose. The distribution and amounts of the three main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens) were determined by dot-blot hybridization using specific 16SrRNA-targeting probes. Enzyme activities were higher in the rumen than in the cecum and before feeding than at 3 h after feeding. The sum of the three cellulolytic bacterial species represented, on average, 4.5% of the total bacterial RNA in the two compartments and did not vary with sampling time. The cellulolytic bacterial community structure was different in the two compartments, with F. succinogenes as the main species in the rumen and R. flavefaciens in the cecum. The lower cellulolytic activity in the cecum than in the rumen could not be ascribed to any difference in the structure of the cellulolytic bacterial community between these two compartments, and other hypotheses related to digestion are proposed. PMID- 11890417 TI - Effect of corn processing on starch digestion and bacterial crude protein flow in finishing cattle. AB - Six ruminally and duodenally cannulated yearling steers (523 kg) were used in a replicated 3 x 3 Latin square design experiment to study the effects of corn processing on nutrient digestion, bacterial CP production, and ruminal fermentation. Dietary treatments consisted of 90% concentrate diets that were based on dry-rolled (DRC), high-moisture (HMC), or steam-flaked (SFC) corn. Each diet contained 2.0% urea (DM basis) as the sole source of supplemental nitrogen. Each period lasted 17 d, with d 1 through 14 for diet adaptation and d 15 through 17 for fecal, duodenal, and ruminal sampling. Dry matter and OM intakes were similar for DRC and SFC but were approximately 15% higher (P < 0.05) for HMC. True ruminal OM digestibilities were 18 and 10% greater (P < 0.05) for HMC than for DRC or SFC, respectively. Ruminal starch digestibilities were similar between HMC and SFC and were approximately 19% greater (P < 0.05) than DRC. Postruminal OM digestibility was similar among treatments; however, postruminal starch digestibility was 15% greater (P < 0.05) for SFC than for DRC or HMC, which were similar. Total-tract DM and OM digestibilities were similar between HMC and SFC and were 4% greater (P < 0.05) than DRC. Likewise, total-tract starch digestibilities were similar between HMC and SFC and were 3% greater (P < 0.05) than DRC. Bacterial CP flow to the duodenum was 29% greater (P < 0.05) for HMC than for DRC or SFC, which were similar. Bacterial N efficiencies were similar among treatments. Based on bacterial CP flow from the rumen, we estimate that dietary DIP requirements are approximately 12% higher for HMC-based diets than for DRC or SFC-based diets, which were similar. PMID- 11890418 TI - Impact of EasiFlo cottonseed on feed intake, apparent digestibility, and rate of passage by goats fed a diet containing 45% hay. AB - EasiFlo cottonseed (ECS), produced by coating whole cottonseed (WCS) with cornstarch to simplify handling and mixing with other ingredients, is marketed commercially. The objective of this trial was to determine its digestibility by small ruminants. Four mature Nubian wether goats, in a 4 x 4 Latin square arrangement of treatments, were fed diets that contained about 45% bermudagrass hay (BGH) plus 0, 15.7, 32.7, or 50.3% ECS, with the ECS replacing corn and soybean meal (2:1 ratio) in the concentrate portion of the diet. Feed intakes and digestibility of components were measured, and passage rate was estimated using ytterbium-marked BGH. Dry matter intake decreased at an increasing rate (P < 0.01) as ECS or fat concentration in the diet increased. Digestibility was linearly depressed (P= 0.003) as ECS replaced corn and soybean meal in the diet, primarily due to depressed (P < 0.05) digestibility of NDF, ADF, and nonfibrous carbohydrates (NFC). In contrast, fat digestibility tended to increase (P = 0.11) linearly and N utilization was increased (P = 0.04) linearly as ECS concentration was increased. Passage kinetics were not altered. Based on regression estimates of TDN for BGH and literature values of TDN for other feed ingredients, the total digestible nutrient content of ECS for mature goats fed a 45% roughage diet was estimated to be 78.0 +/- 9.1%, a value quite similar to that (77.2%) proposed for whole cottonseed for large ruminants by NRC tables. Low digestibility of fiber (under 10% of NDF) and of nonfibrous carbohydrate limits the digestibility of DM and energy from ECS. PMID- 11890419 TI - Effects of restricted and ad libitum intake of diets containing wheat middlings on site and extent of digestion in steers. AB - A 5 x 5 Latin square design was used to determine the effects of restricted and ad libitum intake of diets containing wheat middlings on the site and extent of digestion compared to ad libitum intake of a corn-based diet and ad libitum intake of chopped alfalfa hay. Five ruminally and duodenally cannulated Angus steers (519 +/- 41.5 kg BW) were used to compare five dietary treatments. The five treatments were as follows: ad libitum access to a corn-based finishing diet (control), the control diet with 25 percentage units of the corn and soybean meal replaced with wheat middlings offered ad libitum (WM), the WM diet restricted to 75% of predicted ad libitum intake (RWM), the RWM diet with wheat middlings replaced with ammoniated wheat middlings (RNWM), and ad libitum access to a chopped alfalfa hay diet. Although RWM steers were fed to consume 75% of ad libitum intake, RWM steers consumed 15.5% less DM than WM. Steers fed ad libitum hay consumed 28.6, 31.7, and 37.2% less (P < 0.01) DM, OM, and nitrogen than RWM steers. No differences in apparent or true ruminal digestibility were observed among steers fed the control vs WM, WM vs RWM, RWM vs RNWM, or RWM vs hay diets. However, the steers fed the hay diet had 32.5, 33.4, and 36.9% lower (P < 0.01) apparent total tract digestibilities of DM, OM, and N than those fed the RWM diet. Average ruminal pH was lower (P < 0.01) for control steers than those fed the WM diet and for those fed RWM compared to the hay diet. The acetate:propionate ratio was higher for cattle fed hay vs the RWM diet. Microbial DM and OM flow to the small intestine was higher (P < 0.02) for steers fed the RWM diet than those fed the hay diet. In addition, bacterial N flow to the small intestine was higher (P < 0.01) for cattle receiving the RWM diet than the hay diet. Feeding diets containing 25 percentage units of wheat middlings at 75% ad libitum intake had no effect on ruminal digestibility. PMID- 11890420 TI - Dried poultry waste for cows grazing low-quality winter forage. AB - Two trials conducted in 1996-97 measured BW and body condition score changes of cows fed different sources of degradable intake protein, including dried poultry waste and soybean meal, while grazing low-quality winter forages. In Trial 1, 60 spring-calving cows (5 yr; 555 kg) were used in an individual supplementation trial. Cows were gathered three times a week, sorted into individual pens, and fed their respective supplement. Cows grazed dormant native Sandhills winter range (common pasture) and were assigned to one of six supplemental treatments: 1) no supplement, 2) urea, 3) 22% dried poultry waste + urea, 4) soybean meal, 5) 22% dried poultry waste + soybean meal, or 6) 44% dried poultry waste. All supplements were based on wheat middlings and soybean hulls and were formulated to contain 44% CP. Thirty-six cows were selected randomly (six per treatment) for a 5-d measurement of forage intake from December 16 through December 20, 1996. Cows receiving supplements gained more weight (P < 0.001) and maintained greater body condition (P < 0.001) than unsupplemented cows. Cows receiving urea gained less (P < 0.10) than cows receiving a source of natural protein, but body condition remained similar. No differences were found in daily forage or total organic matter intake (P > 0.10). In Trial 2, cows grazed corn residues. Forty eight spring-calving cows were group-fed supplements in one of six 4-ha paddocks. Cows received supplements containing either soybean meal or dried poultry waste that were the same as the soybean meal and 44% dried poultry waste supplements fed in Trial 1; gains were not different (P > 0.10). Under the economic conditions that existed at the time of these experiments, the supplement containing dried poultry waste resulted in a savings of $.04 per cow per day and a total savings of $3.20 per cow over an 80-d period. Feeding a supplement containing dried poultry waste resulted in performance similar to that when feeding a more conventional supplement containing soybean meal. PMID- 11890421 TI - Relationships among heat production, body weight, and age in Finnsheep and Rambouillet ewes. AB - It was the objective of this study to quantify heat production across ages of Rambouillet and Finnsheep ewes and to evaluate the previous hypothesis that breed differences can be accounted for by scaling for proportion of mature body weight. Seventy-two Finnsheep and 55 Rambouillet ewes were sampled. Heat production was estimated based on individual animal gaseous exchange, which was determined from 55 through 71 h of the feed restriction. Heat production per unit BW decreased as sheep aged, and the breed-specific functions fit the data better than the pooled functions. The rate of decrease in heat production was greater in Finnsheep ewes until 37 wk of age. The rate of growth of Rambouillet ewes was greater than that of Finnsheep ewes over the first 52 wk of age, and Rambouillet ewes reached 95% of their mature BW at an earlier age (71 wk) than did Finnsheep ewes (113 wk). At any given age, Rambouillet ewes had achieved a greater proportion of their mature BW and had a lower heat production per unit BW than Finnsheep ewes. This study demonstrated the necessity of accounting for both age and breed when estimating metabolic rate in sheep. Furthermore, this study suggested that breed and age differences in metabolic rate could be accounted for by scaling for proportion of mature BW and that daily heat production per unit BW (kcal/kg) of Finnsheep, Rambouillet, Suffolk, and Texel ewes can be described by the function /(BW, matBW) = 59.5e(-0.797(Bw/matBw)), where BW = body weight and matBW = mature body weight. PMID- 11890422 TI - Neutral detergent fiber concentration of corn silage and rumen inert bulk influences dry matter intake and ruminal digesta kinetics of growing steers. AB - Corn silage with high NDF concentration has the potential to reduce DMI because it has a greater filling effect in the rumen than low-NDF corn silage. Our objective was to determine whether ruminal fill influences DMI to the same extent with low- or high-NDF corn silage-based diets. Eight ruminally cannulated Holstein steers (198 +/- 13 kg) were randomly assigned to a 2 x 2 factorial arrangement of treatments in a replicated 4 x 4 Latin square design with 16-d periods. Treatments were diets containing corn silage from a normal hybrid (low fiber; LF) or its male-sterile isogenic counterpart (high-fiber; HF), offered for ad libitum consumption to steers with or without rumen inert bulk (RIB). The LF and HF diets contained 33.8 and 50.8% dietary NDF, respectively. Rumen inert bulk was added at 25% of pretrial ruminal volume in the form of plastic-coated tennis balls filled with sand to achieve a specific gravity of 1.1 and a total volume of 7.5 L. No fiber level x inert bulk interactions were detected for DMI or NDF intake (P > 0.10), suggesting that DMI was limited to the same extent by physical fill at both levels of dietary fiber. Addition of RIB decreased DMI by an average of 10.7%, which was 65.5 g/L of added bulk. The HF diet depressed DMI by an average of 15.5%, increased NDF intake 27.1%, and reduced ruminal NDF turnover time by 21.0% compared to the LF diet (P < 0.01), with no effect on ruminal volume or amount of NDF in the rumen (P > 0.10). Addition of RIB also reduced ruminal NDF turnover time and amount of NDF in the rumen (11.8% and 20.7%, respectively; P < 0.01), with no change in ruminal digesta volume (P > 0.10). The HF treatment decreased digestibility of DM and GE (5.5 and 5.7%, respectively; P < 0.01) but increased NDF digestibility (10.4%; P < 0.01) compared to LF. Rumen inert bulk had no effect on digestibility of DM, NDF, or GE (P > 0.10). The lack of reduction in digesta volume with addition of inert fill suggests that DMI of light-weight steers receiving corn silage-based diets within a wide range of NDF concentrations was not regulated by ruminal distension alone. PMID- 11890423 TI - Neutral detergent fiber concentration in corn silage influences dry matter intake, diet digestibility, and performance of Angus and Holstein steers. AB - Twelve Angus (237 +/- 13 kg) and twelve Holstein (235 +/- 15 kg) steers were used to determine whether corn silage-based diets with different NDF levels influence DMI to a similar extent in Angus and Holstein steers and as body weight of the steers increase. Steers were randomly assigned to individual slatted-floor pens and used in a crossover design consisting of six 14-d periods. Experimental diets contained corn silage from a normal hybrid (low-fiber; LF) and its male-sterile counterpart (high-fiber; HF) and were alternated each period. The LF and HF diets contained 33.8 and 50.8% NDF, respectively. The HF diet decreased (P < 0.01) overall steer mean DMI 14.0% relative to LF, with mean differences increasing as steers increased in BW (P < 0.01). Feeding the HF diet also reduced ADG by an average of 13.8% relative to the LF diet (P < 0.01). Holstein steers consumed 14.4% more DM and gained 14.3% faster (P < 0.01) than Angus steers. There was a fiber level x breed-type interaction (P = 0.08) for efficiency of gain. Angus steers receiving the HF diet had greater efficiency of gain than Angus steers consuming the LF diet; however, Holstein steers consuming the LF diet had greater efficiency of gain than those receiving the HF diet. The HF treatment reduced total-tract digestibility of DM and GE by 4.6 and 4.5%, respectively (P < 0.01), and decreased DE intake by 20.5% (P < 0.01) but increased apparent totaltract digestibility of NDF and ADF (9.4 and 8.4%, respectively; P < 0.01). Holstein steers had similar digestibility of DM and GE (P > 0.10) but had greater DE intake (P < 0.01) compared to Angus steers. There were fiber level x breed-type interactions for total-tract digestibility of NDF and ADF (P < 0.06). The difference in DM digestibility was negatively associated with the difference in DMI (r2 = 0.23; P < 0.01) for LF minus HF within Angus steers, but not within Holstein steers (P = 0.42). Total-tract digestibility of NDF and ADF was 4.1 and 3.4% lower for the HF diet but was only 1.1 and 0.6% lower for the LF diet when fed to Holstein compared to Angus steers. Results from this trial demonstrate that high-NDF corn silage-based diets reduced intake of both Angus and Holstein steers, and this reduction in DMI continued as steers increased in BW from 235 to 330 kg. Breed differences were also noted for digestible energy intake as influenced by fiber level. PMID- 11890424 TI - Concentrations of conjugated linoleic acid (cis-9, trans-11-octadecadienoic acid) are not increased in tissue lipids of cattle fed a high-concentrate diet supplemented with soybean oil. AB - Conjugated linoleic acid (CLA), a mixture of isomers of linoleic acid, has many beneficial effects, including decreased tumor growth in animal cancer models. The cis-9, trans-11 isomer of CLA (CLA9,11) can be formed in the rumen as an intermediate in biohydrogenation of linoleic acid. Recent data, however, indicate that tissue desaturation of trans-fatty acids is an important source of CLA9,11 in milk. Our objective was to determine whether supplementing a high-corn diet with soybean oil (SBO; a source of linoleic acid) would increase concentrations of CLA in ruminal contents and tissue lipids. Four ruminally cannulated steers were utilized in a Latin square design with 28-d periods. A control diet (80% cracked corn, 2.0% corn steep liquor, 8.0% ground corn cobs, and 10% supplement [soybean meal, ground shelled corn, minerals, and vitamins]) was supplemented with 2.5, 5.0, or 7.5% (DM basis) SBO. Supplemental SBO did not affect ruminal pH or concentrations of the major VFA. The proportion and amount (mg FA/g DM ruminal contents) of CLA9,11 were not increased by increasing dietary SBO. However, the proportion and amount of the trans-10, cis-12 CLA isomer (CLA10,12) in ruminal contents increased linearly (P < 0.006) as dietary SBO increased. Trans-18:1 isomers in ruminal contents increased linearly (P < 0.02) as dietary SBO increased. The proportion of CLA10,12 was correlated positively (P < 0.001) with proportions of trans-C 18:1 isomers in ruminal contents. Conversely, CLA9,11 was correlated negatively (P < 0.05) with the proportions of trans-18:1 in ruminal contents. The same high-corn diet, supplemented with 0 or 5% SBO, was fed to 20 Angus-Wagyu heifers for 102 d in a randomized complete block design to determine the effect of added SBO on tissue deposition of CLA. Supplemental SBO did not affect feed intake, gain:feed, or carcass quality. Tissue samples were obtained from the hindquarter, loin, forequarter, liver, large and small intestine, and subcutaneous, mesenteric, and perirenal adipose depots. The concentration of CLA9,11 was greatest in subcutaneous adipose tissue but was not affected in any tissue by SBO. Supplementing high-corn diets with SBO does not increase CLA9,11 concentrations in tissues of fattening heifers. Research is needed to identify regulatory factors for pathways of biohydrogenation that lead to increased concentrations of CLA10,12 in ruminal contents when high-oil, high-concentrate diets are fed. PMID- 11890425 TI - Use of critical interactive thinking exercises in teaching reproductive physiology to undergraduate students. AB - In higher education, increasing emphasis is being placed on the use of new technologies in the classroom. However, the emphasis needs to be placed on methods that truly enhance understanding and knowledge retention. Class discussions help students understand and retain information previously presented in lecture format. Furthermore, if students are challenged to critically evaluate, communicate, and defend their ideas, knowledge retention and understanding will increase even more. Critical interactive thinking exercises (CITE) were employed at two different universities to enhance student knowledge retention and promote the development of critical thinking. Applicability of CITE to undergraduate learning was assessed over a 3-yr period in the undergraduate reproductive physiology courses at Michigan State University and the University of Missouri-Columbia. For each exercise, students were challenged to prepare a one-page, double-spaced composition addressing an incompletely understood phenomenon or problem-solving situation related to the reproductive system. In preparing their compositions, students were encouraged to use information previously presented in lecture plus outside information to develop their ideas. Students were required to formulate and defend a hypothesis or approach to the problem presented. At the subsequent class period, students were divided into groups of three to four, in which they interactively discussed their ideas. Each group member was challenged to defend his or her hypothesis and explanation and to persuade other group members to adopt their ideas. Each group then arrived at a consensual opinion that was presented during a discussion by the entire class. The class then debated the merits of each group's hypothesis or explanation and the supporting arguments presented. At first, the students were apprehensive about the CITE, particularly about communicating and defending ideas with their classmates. However, course evaluations showed that 131 out of 137 students considered the CITE a positive experience that enhanced learning. Additionally, 131 out of 137 students reported that the CITE enhanced their critical thinking skills. We feel that the use of CITE in teaching reproductive theory to undergraduate students fosters critical thinking skills, communication skills, and knowledge retention. The general concept can be readily applied to courses in other subject areas in the animal sciences. PMID- 11890427 TI - Rapid communication: linkage mapping of a microsatellite isolated from a BAC clone containing the protein kinase C binding protein 2 on bovine chromosome 1. PMID- 11890426 TI - Rapid communication: partial nucleotide sequence of the goat stearoyl coenzyme A desaturase cDNA and gene structure. PMID- 11890428 TI - Acupuncture in the twenty-first century. PMID- 11890430 TI - Issues in planning a placebo-controlled trial of manual methods: results of a pilot study. AB - OBJECTIVE: There are fundamental differences between the administration of medications and the application of manual procedures, such as those used by chiropractors. The objective of this study was to gather preliminary information on how to address these differences in the design of a multisite, randomized placebo-controlled trial of chiropractic care for women with chronic pelvic pain (CPP). DESIGN: Pilot study for a multisite, randomized, placebo-controlled clinical trial. SETTING: Three chiropractic research clinics in the midwest United States. SUBJECTS: Thirty-nine (39) women with CPP of at least 6 months' duration, diagnosed by board-certified gynecologists. INTERVENTIONS: The active intervention consisted of the chiropractic technique, lumbar spine flexion distraction, combined with manual Trigger Point Therapy. The placebo intervention consisted of a sham chiropractic procedure performed with an instrument combined with effleurage (light massage). OUTCOME MEASURES: The primary outcome measure was the change in the Pain Disability Index (PDI) from baseline to the end of treatment (6 weeks), assessed by group and site. If the change score was in the same direction at all sites, the results were to be combined to estimate treatment effect size. RESULTS: Patient characteristics were similar to those of patients with CPP in other studies. Recruitment methods, particularly in respect to the eligibility criteria and screening protocols, would require modification in order to recruit an adequate sample for the planned randomized controlled trial. Clinicians followed standardized procedures with apparently minimal deviation, patients in both groups were satisfied with their care and blinding appeared to be successful. PDI change scores were not consistent across sites and so results were not combined and overall treatment effect sizes were not estimated. CONCLUSIONS: The technical and personnel resources required to achieve adequate standardization of procedures at multiple sites may make a placebo controlled trial unfeasible, given our current lack of knowledge about the active agent in manual chiropractic procedures. PMID- 11890429 TI - Description and validation of a noninvasive placebo acupuncture procedure. AB - OBJECTIVE: To evaluate a simulated acupuncture technique for use in randomized controlled trials assessing the efficacy of acupuncture for low-back pain. SETTING: The clinic of an accredited acupuncture college in Seattle, Washington. SUBJECTS: Acupuncture-naive enrollees of Group Health Cooperative who had visited their primary care provider with a complaint of back pain that persisted for at least 3 months. EXPERIMENTAL DESIGN: In the first experiment, subjects received six insertions of real needles and six pokes with a toothpick in a guidetube in a two-period crossover design. In the second experiment, subjects were randomly assigned to receive either a complete treatment with real acupuncture needles or a simulated treatment using a toothpick in a guidetube. OUTCOMES: In the first experiment, we compared subjects' perceptions about which implement was used for each "insertion" while in the second, we compared the perceptions (e.g., acupuncturist's warmth and caring, the reasonableness of acupuncture as a treatment) and pain relief of those who received an acupuncture treatment using needles to those receiving simulated acupuncture. RESULTS: In the first experiment, the toothpick insertions were perceived as slightly more like real needling than the real needling (mean ratings of 2.8 and 2.1, respectively; p = 0.08). In the second experiment, 52% percent of those receiving the simulated needling versus 65% of those receiving real acupuncture believed they were "definitely" or "probably" receiving real acupuncture (p = 0.33). Perceptions of acupuncture, as measured by a credibility questionnaire, were similar in the two groups. Those receiving real acupuncture were more likely to report immediate pain relief, and this was the factor most predictive of the subject's belief about which treatment they had received (p = 0.02). CONCLUSIONS: The simulated acupuncture procedure evaluated in this study represents a reasonable control treatment for acupuncture-naive individuals in randomized controlled trials assessing the efficacy of acupuncture for low-back pain. PMID- 11890431 TI - Homeopathy and infectious disease: controversies raised by the recent foot-and mouth disease and anthrax outbreaks. PMID- 11890432 TI - The clinical effectiveness of healing touch. AB - OBJECTIVES: (1) to determine the clinical effectiveness of Healing Touch (HT) on variables assumed to be related to health enhancement; (2) to determine whether practitioner training level moderates treatment effectiveness. DESIGN: Mixed method repeated measures design with quasi-experimental and naturalistic approaches, paired with nomothetic and idiographic analyses. SETTING/LOCATION: Practitioner's offices or client's home. SUBJECTS: Twenty-two (22) clients who had never experienced HT. INTERVENTIONS: Three treatment conditions: no treatment (NT), HT only (standard HT care), and HT+ (Standard HT care plus music plus guided imagery). OUTCOME MEASURES: Secretory immunoglobulin A (sIgA) concentrations in saliva, self-reports of stress levels, client perceptions of health enhancement, and qualitative questionnaires about individual effects. RESULTS: Clients of practitioners with more training experienced statistically significant positive sIgA change over the HT treatment series, while clients of practitioners with less experience did not. Clients reported a statistically significant reduction of stress level after both HT conditions. Perceived enhancement of health was reported by 13 of 22 clients (59%). Themes of relaxation, connection, and enhanced awareness were identified in the qualitative analysis of the HT experience. Pain relief was reported by 6 of 11 clients (55%) experiencing pain. CONCLUSIONS: The data support the clinical effectiveness of HT in health enhancement, specifically for raising sIgA concentrations, lowering stress perceptions and relieving pain. The evidence indicates that positive responses were not exclusively as a result of placebo, that is, client beliefs, expectations, and behaviors regarding HT. PMID- 11890433 TI - The effect of immunization with killed tumor cells, with/without feeding of Echinacea purpurea in an erythroleukemic mouse model. AB - OBJECTIVE: Tumor amelioration via vaccination/immunization is a practice for which considerable experimental and clinical support is growing. Combination therapies have proven to be more beneficial than treatment with single agents. We hypothesized that immunization of mice with killed erythroleukemia cells prior to the induction of erythroleukemia via injection of viable tumor cells, plus dietary administration of a known immuno-enhancing phytocompound, Echinacea purpurea, would be more effective than immunization alone. DESIGN: A commercially available extract of E. purpurea root, already proven as a natural killer (NK) cell stimulant, was administered via the chow, for periods of 9 days or 3 months after the onset of leukemia to mice which had been injected (immunized) 5 weeks earlier with killed leukemia cells. RESULTS: Immunized mice (+/- E. purpurea) had significantly prolonged life spans versus non-immunized mice, with an even greater proportion of hosts surviving long-term in the E. purpurea-fed group. NK cells, the mediators of nonspecific immunity and well-demonstrated mediators of tumor cytolysis, were very significantly elevated in immunized, leukemic mice receiving E. purpurea in their diet versus those receiving untreated chow. Early in tumor development (9 days), cells mediating specific immunity (T, B lymphocytes) were 10-12 times higher in absolute numbers in the spleens in all immunized, leukemic mice vs unimmunized, leukemic mice at the same stage of tumor progression. CONCLUSIONS: The results demonstrate that combination therapy, involving specific tumor cell immunization, followed by daily phytotherapy (dietary E. purpurea), sensitized the immune cells and led to life span prolongation greater than that provided by immunization alone. PMID- 11890434 TI - Methodological changes in the evaluation of complementary and alternative medicine: issues raised by Sherman et al. and Hawk et al. PMID- 11890435 TI - Opinions and practices of medical rehabilitation professionals regarding prayer and meditation. AB - OBJECTIVE: To assess the attitudes and practices of professionals in the field of physical medicine and rehabilitation (PM&R) regarding prayer and meditation. DESIGN: A national mail survey that included questions about the use of a number of complementary and alternative therapies. PARTICIPANTS: The survey was mailed to 7,479 physicians, nurses, physical therapists, and occupational therapists who specialize in PM&R, and 1221 (17%) returned completed surveys. RESULTS: Although the majority of respondents endorsed prayer as a legitimate health care practice, there was greater belief in the benefits of meditation. Older respondents were more likely to recommend meditation to their patients and more likely to meditate themselves. Gender differences that were observed in opinions and practices are better interpreted as differences in professional specialty. In general, nurses and occupational therapists responded more positively toward meditation and prayer than did physicians and physical therapists. Personal use of a technique was the strongest predictor of professional behaviors. Attitude was a stronger predictor of professional use or referral for prayer than meditation, but correlations between attitude and behavior were generally weak for both techniques. Despite their acceptance of these techniques, the vast majority of rehabilitation professionals did not refer their patients for meditation or religious consultation. CONCLUSIONS: Although there were significant relationships among beliefs, and personal and professional behaviors regarding these techniques, a large part of the variance in professional behaviors was not accounted for by age, gender, opinion, or personal behavior, indicating that other influences exert a stronger effect on professional practice decisions. PMID- 11890436 TI - Building CAM databases: the challenges ahead. PMID- 11890437 TI - Effects of a very low-fat, vegan diet in subjects with rheumatoid arthritis. AB - OBJECTIVE: To demonstrate the effects of a very low-fat, vegan diet on patients with rheumatoid arthritis (RA). DESIGN: Single-blind dietary intervention study. SUBJECTS AND STUDY INTERVENTIONS: This study evaluated the influence of a 4-week, very low-fat (approximately 10%), vegan diet on 24 free-living subjects with RA, average age, 56 +/- 11 years old. OUTCOME MEASUREMENTS: Prestudy and poststudy assessment of RA symptomatology was performed by a rheumatologist blind to the study design. Biochemical measures and 4-day diet data were also collected. Subjects met weekly for diet instruction, compliance monitoring, and progress assessments. RESULTS: There were significant (p < 0.001) decreases in fat (69%), protein (24%), and energy (22%), and a significant increase in carbohydrate (55%) intake. All measures of RA symptomatology decreased significantly (p < 0.05), except for duration of morning stiffness (p > 0.05). Weight also decreased significantly (p < 0.001). At 4 weeks, C-reactive protein decreased 16% (ns, p > 0.05), RA factor decreased 10% (ns, p > 0.05), while erythrocyte sedimentation rate was unchanged (p > 0.05). CONCLUSION: This study showed that patients with moderate-to-severe RA, who switch to a very low-fat, vegan diet can experience significant reductions in RA symptoms. PMID- 11890439 TI - Standards for Reporting Interventions in Controlled Trials of Acupuncture: the STRICTA recommendations. AB - Acupuncture treatment and control group interventions in parallel-group randomized trials of acupuncture are not always precisely reported. In an attempt to improve standards, an international group of experienced acupuncturists and researchers devised a set of recommendations, designating them STRICTA: STandards for Reporting Interventions in Controlled Trials of Acupuncture. In a further consensus-building round, the editors of several journals helped redraft the recommendations. These follow the Consolidated Standards for Reporting Trials (CONSORT) format, acting as an extension of the CONSORT guidelines for the specific requirements of acupuncture studies. Participating journals are publishing the STRICTA recommendations and requesting prospective authors to adhere to them when preparing reports for publication. Other journals are invited to adopt these recommendations. The intended outcome is that interventions in controlled trials of acupuncture will be more adequately reported, thereby facilitating an improvement in critical appraisal, analysis and replication of trials. PMID- 11890440 TI - Homeopathic physician with his vials of medications. PMID- 11890438 TI - Comparison of propolis skin cream to silver sulfadiazine: a naturopathic alternative to antibiotics in treatment of minor burns. AB - BACKGROUND: Propolis, a naturopathic substance derived from bees wax extract, has recently been praised for its antimicrobial, anti-inflammatory, and cicatrization enhancing properties. OBJECTIVE: In our study, we compare these properties in a high-grade Brazilian propolis skin cream directly with silver sulfadiazene (SSD) in the treatment of minor burns (superficial second degree) in the ambulatory care setting (less than 20% total body surface area burned). SETTINGS/LOCATION: The study was conducted at the burn clinic in Pronto Socorro para Queimaduras, Gioania, Brazil. SUBJECT: Patients were admitted to the study only if their initial presentation for burn care was within 48 hours postinjury and if bilateral wounds of similar depth and quality were present. INTERVENTIONS: Patients had propolis skin cream applied to one wound and SSD applied to the other selected wound on initial presentation and underwent debridement and dressings change the following morning. Patients subsequently returned to the clinic every 3 days to have the wounds checked and dressings changed. At these check-ups, wounds were cultured for microbial growth and photographed to document inflammation and cicatrization. Patients were instructed not to disturb their wounds or change their dressings at home, thus propolis skin cream and SSD were applied to the wounds only at the specified 3-day intervals. RESULTS: Our preliminary results do not show any significant difference in microbial colonization between wounds treated with SSD and propolis skin cream, however, wounds treated with propolis skin cream consistently showed less inflammation and more rapid cicatrization then those treated with SSD. CONCLUSION: Propolis skin cream appears to have a beneficial effects on the healing of partial thickness burn wounds. If dressings had been changed more frequent the antimicrobial and wound healing effects would have been enhanced. PMID- 11890441 TI - Report from Asia: what is new in the field of microcirculation. PMID- 11890442 TI - Surveys of complementary and alternative medicine: Part V. Use of alternative and complementary therapies for psychiatric and neurologic diseases. AB - Surveys of general complementary and alternative medicine (CAM) use have suggested an association with high levels of depression and anxiety. This raises the question of whether anxious or depressed people seek CAM, or whether there are underlying factors associated with long-term chronic illness. There is no clear indication from four surveys of psychiatric patients. These are summarized and presented. A separate table summarizes three studies of patients with neurologic diseases, two of patients with multiple sclerosis, and one of a mixed patient population. More studies are amassing in specific disease areas, although it is difficult to detect clear trends because of methodological and terminological incompatibilities. PMID- 11890443 TI - Flank position ureterorenoscopy: new positional approach to aid in retrograde caliceal stone treatment. AB - BACKGROUND AND PURPOSE: Clearance of lower pole stones and complex maneuvers such as basket repositioning remain challenging in ureteroscopic treatment of caliceal stone burdens. We describe the initial use of flank position ureteroscopy (FPU) with modifications to aid in the treatment of complex caliceal urolithiasis. We hypothesized that gravitational force acting on caliceal stones and fragments during holmium laser treatment would result in fragments being repositioned into the dependent renal pelvis, allowing enhanced treatment. PATIENTS AND METHODS: Eleven patients with complex upper tract stone disease (aggregate stone burden 1.5-5 cm) were treated using FPU. Patients were positioned with the stone containing side superior. Flexible cystoscopy followed by flexible ureterorenoscopy and intracorporeal lithotripsy were performed. We utilized several modifications of the technique, including a radiolucent table and rotational C-arm fluoroscopy unit. Once all stones and particles fell into the dependent renal pelvis, additional laser fragmentation was simple. RESULTS: We observed the expected advantage of gravitational drainage of particles and stone into the renal pelvis during procedures. In many cases, fragments passed partially down the ureter during treatment. Basket repositioning of stones was not necessary. One complication, collecting system perforation during access, was treated with stent placement and successful delayed FPU. Seven patients were stone free at follow-up. Four patients with stone burdens >3 cm had approximately 80% stone burden reduction, but residual asymptomatic lower-pole particles (2-6 mm) remained, with several clearing with a secondary procedure. CONCLUSIONS: The use of FPU for the performance of complex renal stone treatment is a useful aid. Further refinement and comparison with standard technique is ongoing. PMID- 11890444 TI - Ureteroscopy as a first-line intervention for ureteral calculi in pregnancy. AB - BACKGROUND AND PURPOSE: Renal colic in pregnancy presents a diagnostic and therapeutic challenge. When conservative therapy fails or is not indicated, temporary measures such as ureteral stenting are often chosen as a first-line intervention, postponing definitive management until after delivery. We propose that advances in endoscopic equipment and anesthesia techniques dictate a more definitive strategy. PATIENTS AND METHODS: A retrospective analysis was performed on 10 consecutive pregnant patients presenting with renal colic necessitating intervention between April 1998 and April 2000. The mean patient age was 23 (range 17-31) years. One patient presented during the first trimester, six in the second, and three in the third. Four of the patients had a history of stone disease. All patients had flank pain at presentation, six on the left side and four on the right. Hematuria, fever, and nausea were present in eight, one and two patients, respectively. RESULTS: Ultrasound scanning was performed in all patients and showed a low sensitivity (28.5%) when compared with intraoperative findings. Ureteroscopy (rigid and/or flexible) was performed as a first-line intervention in six patients, in two of whom no stone was found. Percutaneous nephrolithotomy was carried out in one patient presenting with a nephrostomy tube. Double-J stents were placed in only three patients with specific indications, namely urinary infection, late gestational phase, and difficult ureteroscopy secondary to a narrow ureter. No obstetric or urologic complications were noted. The mean size of the stones retrieved in seven patients was 7 mm. CONCLUSIONS: Ureteroscopy may be considered a safe and effective first-line definitive therapeutic option in pregnant patients requiring intervention for stone disease. PMID- 11890445 TI - Laparoscopic radical nephrectomy: prospective assessment of impact of intact versus fragmented specimen removal on postoperative quality of life. AB - PURPOSE: We prospectively compared postoperative recovery and quality of life for groups of patients undergoing laparoscopic radical nephrectomy with intact or fragmented specimen removal. PATIENTS AND METHODS: A prospective evaluation of 12 patients having a transperitoneal laparoscopic nephrectomy was completed. In each case, a radical dissection was performed regardless of the surgical indication. Fragmented specimens (N = 7) were extracted at the umbilical port, and intact specimens (N = 5) were extracted through an infraumbilical incision. Demographic and perioperative data including specimen removal incision, narcotic requirements, and recovery interval were recorded. Subjective pain and activity assessments were administered prospectively on postoperative days 1, 2, 7, and 14. RESULTS: The mean incision length for intact specimen removal was 7.6 cm and that for fragmented removal was 1.2 cm (P < 0.05). Pain and activity self assessments improved over time in each group. No significant differences in pain or activity scores were noted between treatment groups at any queried interval. Time to return of normal activity was not significantly different in the two groups. CONCLUSIONS: In this pilot study, no subjective or objective advantage was demonstrated for kidney fragmentation during laparoscopic radical nephrectomy. A larger randomized study is required to better assess any clinical advantage to specimen morcellation. PMID- 11890446 TI - Ureteral segmental replacement using multilayer porcine small-intestinal submucosa. AB - PURPOSE: To assess the outcome of segmental ureteral replacement using a new multilayer porcine small-intestinal submucosa (SIS), Surgisis ES (Cook Inc., Stouffville, ON, Canada) designed to provide enhanced strength. MATERIALS AND METHODS: The ureters of five female farm pigs were accessed through a median laparotomy incision. A segment of 2-cm midureter was resected bilaterally. The left ureteral segments were replaced by 10F tubularized SIS segments using 5-0 PDS interrupted sutures. The right ureters were primarily end-to-end anastomosed, serving as controls. Internal pigtail stents were left bilaterally for 6 weeks. One animal at 3 weeks, one animal at 6 weeks, and three animals at 12 weeks were sacrificed. The patency of the ureters was assessed by retrograde pyelography at 6 and 12 weeks, while inflammation and regeneration were assessed grossly and histologically. RESULTS: At 3 and 6 weeks, both experimental and control ureters were patent without extravasation on retrograde studies. Adhesions and signs of ureteral inflammation were found only on the SIS side. The graft was partially and completely epithelialized at 3 and 6 weeks, respectively. However, at 12 weeks, all the ureters on the experimental side were completely occluded, while on the control side, all were patent. Although histologically, urothelium and muscular cells had proliferated over the graft, they were embedded in an intense fibrotic and inflammatory process. At 12 weeks, all animals had developed hydroureteronephrosis above the grafts. CONCLUSIONS: Technically, Surgisis ES was easily modeled, providing conditions for a water-tight anastomosis. None of the animals developed urinary fistula. Regeneration of urothelium and muscle were induced and supported by the graft. However, functional replacement was not successful. A suitable material for this purpose has yet to be discovered. PMID- 11890447 TI - The Dornier Compact Delta lithotripter: the first 500 renal calculi. AB - BACKGROUND AND PURPOSE: Extracorporeal Shockwave Lithotripsy (SWL) is now the best noninvasive treatment for renal calculi, rendering many patients stone free. This prospective study was performed to evaluate the short-term results of patients undergoing SWL with the Dornier Compact Delta lithotripter for all renal calculi. PATIENTS AND METHODS: Between April 1999 and May 2000, there were 500 renal calculi treated in 166 female and 334 male patients with a mean age of 53 +/- 15 years. All patients who completed treatment were entered in the study and assessed at 1 and 3 months with a plain film of the kidneys, ureters, and bladder. Stone-free rate and final outcome have been evaluated. Final outcome is defined as stone free or residual fragments 4 mm or less. Analysis has been made according to stone size, location, number of treatments per stone, and number of shocks per stone. The analgesia requirements during each treatment and complications have also been analyzed. RESULTS: The overall stone-free rate for stones <10 mm was 62% at 1 month and 76% at 3 months. For stones 10 to 20 mm, these rates were 53% and 66%, while the rates for stones >20 mm were 41% and 47%, respectively. The final outcome for stones <10 mm was 90% at 1 month and 93% at 3 months, for stones 10 to 20 mm 73% and 84%, and for stones >20 mm 57% and 67%, respectively. The effectiveness quotient for calculi <10 mm was 60%. For calculi 10 to 20 mm, it was 51%, and for those >20 mm, it was 31%. Oral analgesia was given routinely; however, additional intravenous analgesia was necessary in 22% of treatments. No serious complications have been seen. CONCLUSIONS: These results show that with proper patient selection, good results at 1 and 3 months can be achieved with minimal anesthesia during treatment and low retreatment rates. We do not recommend SWL as primary therapy for stones >20 mm. PMID- 11890448 TI - Comparison of 150 simultaneous bilateral and 300 unilateral percutaneous nephrolithotomies. AB - PURPOSE: To compare the results, complications, efficiency, and safety of simultaneous bilateral percutaneous nephrolithotomy (SBPN) and unilateral percutaneous nephrolithotomy (PCNL). PATIENTS AND METHODS: We compared the results and complications of 150 SBPNs with those of 300 unilateral PCNLs. All the procedures were performed by one surgeon which provides relatively constant parameters. The success rates, preoperative and postoperative laboratory results, and complications were compared on the basis of stone size and the number of nephrostomy tracks. RESULTS: There were no significant differences between the results and complications of SBPN and PCNL. The SBPN itself did not cause more blood loss than unilateral PCNL. In both groups, the blood loss was in direct proportion to the size of the stones and the number of nephrostomy tracks. After SBPN, kidney function improved >20% in 12.2% of the patients and worsened for more than 3 days in only 4%. Temporary worsening of kidney function occurred in the unilateral procedure group as well (8%), mostly in cases of solitary kidneys or bilateral stones. The SBPN was not more hazardous than unilateral PCNL (complication rate 11.3% v 14.3%, respectively). In both groups, most of the complications were in proportion to the size and difficulties of the stones. CONCLUSION: Simultaneous bilateral percutaneous nephrolithotomy is a safe and advantageous procedure that is not more hazardous than the separate PCNL in cases of bilateral large stone burdens. To our knowledge, these are the largest reported series of these procedures and the only comparative analysis of SBPN and PCNL. PMID- 11890449 TI - Totally endoscopic management of upper tract transitional-cell carcinoma. AB - BACKGROUND AND PURPOSE: Nephron-sparing therapy arose spurred by efforts to delay dialysis for patients with renal insufficiency or solitary kidneys. As technology has improved, complete endoscopic ablation of tumor via the holmium and Nd:YAG lasers has proven efficacious for cancer control. We have extended ureteroscopic treatment to patients with normal contralateral kidneys given the proper indications. For required extirpative therapy in cases of uncontrolled cancer, laparoscopic nephroureterectomy is rapidly becoming popular and appears to lend the same tumor control as open surgery while significantly lessening morbidity. We reviewed our experience with endourologic treatment and propose an algorithm for the management of upper tract TCC. PATIENTS AND METHODS: Over the period from August 1998 to May 2000, 70 patients underwent ureteroscopic evaluation, treatment, or both for TCC. During the same period, 24 patients had a hand assisted laparoscopic nephroureterectomy (HALNU) performed. A thorough chart review was performed to determine pathologic data and management decision-making. RESULTS: Of the 70 patients evaluated ureteroscopically, 46 were examined for the first time, while the remaining 24 patients were already on the surveillance protocol. Of the 46 initially evaluated patients, 18 were referred for HALNU. Fifteen other patients were placed on surveillance. Of the 24 patients already on surveillance, only 1 required HALNU. The most common reasons for nephroureterectomy were bulky tumors that were ureteroscopically unresectable, high-grade disease, and patient preference. CONCLUSIONS: The combination of ureteroscopy and laparoscopy has made the management of upper tract TCC totally endoscopic, providing decreased morbidity while maintaining cancer control. PMID- 11890450 TI - Transperitoneal laparoscopic renal surgery using blunt 12-mm trocar without fascial closure. AB - BACKGROUND AND PURPOSE: Conical blunt trocar insertion may eliminate the need for fascial closure (FC) in transperitoneal laparoscopic renal surgery. This concept applies to 12-mm blunt trocar placement through muscular parts of the abdominal wall, relying on muscle splitting and eventual muscle retraction when the trocar is removed. We retrospectively assessed the safety of fascial nonclosure (FNC) after 12-mm blunt port insertion. PATIENTS AND METHODS: Ninety transperitoneal laparoscopic renal procedures were performed between August 1999 and May 2000. Four ports (two 12 mm and two 5 mm) were usually used except for 30 donor nephrectomies, where an additional 5-mm port was used. The 12-mm trocars were inserted at the lateral border of the rectus muscle 5 cm below the costal margin and in the anterior axillary line 8 cm below the costal margin. Fascial closure was performed in 62 patients and nonclosure in 28 patients. Exclusion criteria for FNC included midline location, malnutrition, renal failure, and chronic use of steroids. Postoperative outcomes were compared in 20 patients with FNC matched with 20 patients with FC. RESULTS: At an average of 4.8 months of follow-up, none of the patients developed a trocar site hernia. No significant statistical differences were observed between the groups with regard to intraoperative and postoperative data. CONCLUSIONS: These two approaches appear to be equivalent in terms of patient morbidity and postoperative hospital stay. Fascial nonclosure after transperitoneal 12-mm blunt trocar insertion, through muscular parts of the abdominal wall may be safe and efficacious and eliminates the last step in transperitoneal laparoscopic renal surgery. PMID- 11890451 TI - Ureteral ischemia model: an explanation of ureteral dysfunction after chronic obstruction. AB - BACKGROUND AND PURPOSE: Many models of smooth muscle ischemia have been developed to explain organ insufficiency or failure. Ureteral decompensation may also be described in these terms. We anticipate that ureteral ischemia will result from overdistention brought about by obstruction. A preliminary model to create an ischemic ureter is described herein. MATERIALS AND METHODS: Six white New Zealand female rabbits were used for this study. All had their left ureters surgically ligated at the level of the urinary bladder. The right ureters served as controls. In the acute-phase group, the ureters were all reexplored 2 weeks postobstruction. Exploration of the other rabbits was delayed for 3 more weeks. A laser Doppler needle (Transonics Inc.) was used to measure tissue perfusion in the rental artery, renal vein, renal parenchyma, renal pelvis, ureteropelvic junction, upper ureter, mid ureter, lower ureter, and lateral wall of the bladder. Baseline and postobstructive measurements of tissue perfusion were collected and compared. RESULTS: In both the acute and the chronic obstruction groups, there was a demonstrable drop-off in perfusion of the ureteral sidewall. A more notable loss of perfusion was seen in the distal ureter. CONCLUSION: The increased wall tension in the obstructed ureter results in a significant decrease in smooth muscle perfusion. This ischemia may result in the same functional and histologic changes that occur in other smooth muscle organs. Ultimately, the poor outcomes of some restorative/reconstructive procedures on the ureter may be explained in terms of smooth muscle ischemia. PMID- 11890452 TI - Cost effectiveness of treatment for benign prostatic hyperplasia: an economic model for comparison of medical, minimally invasive, and surgical therapy. AB - PURPOSE: To evaluate the cost effectiveness of minimally invasive therapy relative to medical (alpha-blocker) therapy and transurethral resection (TURP) for patients with moderate to severe symptoms of benign prostatic hyperplasia (BPH). METHODS: We constructed a decision-analytic Markov model for a hypothetical cohort of 65-year-old men with moderate to severe BPH symptoms. Microwave thermotherapy was selected to represent minimally invasive treatment. Cost-effectiveness analysis was performed with 25 health states using the 3 treatments, 5 short-term clinical events, and 17 possible long-term outcomes. Each health state had an associated cost and utility. Quality of life (QoL) and utility estimates were obtained by interviewing 13 men with BPH symptoms using the standard gamble reference methods. Patients were classified as risk averse (RA) or non-risk averse (NRA) on the basis of their attitudes to risk. We calculated the incremental cost effectiveness of microwave thermotherapy relative to medical therapy and TURP over 5 years after treatment initiation. Event probabilities were obtained from the literature, a consensus panel, and published randomized clinical trials. RESULTS AND CONCLUSIONS: The utility values generated were internally consistent and externally valid for a hypothetical cohort of 10,000 RA patients. Microwave thermotherapy was preferred by the NRA group, while medical therapy was preferred by the RA group. Surgery was least preferred by both groups. Microwave thermotherapy had a small incremental cost but improved QoL in comparison with medical therapy. Microwave thermotherapy had a higher utility and lower cost than TURP and thus was dominant over TURP. This analytical method can be applied to evaluate the cost effectiveness of any BPH therapy. PMID- 11890453 TI - Routine ureteral stenting is not necessary after ureteroscopy and ureteropyeloscopy: a randomized trial. AB - BACKGROUND AND PURPOSE: Retrospective studies have suggested that routine stenting can be avoided following ureteroscopy. We prospectively analyzed the need for routine ureteral stent placement in patients undergoing ureteroscopic procedures. PATIENTS AND METHODS: Fifty-five consecutive patients (60 renal units) were randomized into either a stent or a no-stent group following ureteroscopy with either a 7.5F semirigid or a 7.5F flexible ureteroscope for treatment of calculi (holmium laser or pneumatic lithotripsy) or transitional cell carcinoma (holmium laser). Intraoperative variables assessed included total stone burden, the need for ureteral dilation, and overall operative times. All patients were evaluated by questionnaire on postoperative days 0, 1, and 6 with regard to pain, frequency, urgency, dysuria, and hematuria. RESULTS: Of the 60 renal units treated, 38 received ureteral stents (mean 5.2 days), and 22 were treated without a stent. All 10 patients requiring ureteral balloon dilation had stents placed and were removed from the analysis. There was no significant difference between the groups with regard to age, sex, or stone burden. Operative time was decreased in the no-stent group (43 minutes v 55 minutes; P = 0.013). Flank discomfort was significantly less common in the no-stent group on days 0, 1, and 6 (P = 0.004, P = 0.003, P < 0.001, respectively), as was the incidence of suprapubic pain on day 6 (P = 0.002). There was no difference in urinary frequency, urgency, or dysuria between the groups on postoperative day 1, but all these symptoms were significantly reduced in the no-stent group on day 6 (P < 0.001, P < 0.001, P = 0.002, respectively). There was no significant difference in patient-reported postoperative hematuria in either group. One patient in each group developed a urinary tract infection. One patient in the no-stent group developed ureteral obstruction in the postoperative period that necessitated stenting, and one patient in the stent group experienced stent migration necessitating removal. CONCLUSIONS: Routine ureteral stenting does not appear to be warranted in those patients who do not require ureteral dilation during ureteroscopic procedures. Ureteral stent placement following ureteroscopy may be avoided, thereby reducing operative time, surgical costs, and patient morbidity. PMID- 11890454 TI - Roles of the complement system in human neurodegenerative disorders: pro inflammatory and tissue remodeling activities. AB - Complement is an important component of the innate immune response with the capacity to recognize and clear infectious challenges that invade the CNS through a damaged blood brain barrier. For instance, the membrane attack complex is involved in cytotoxic and cytolytic activities while other smaller fragments lead to cell activation (chemotaxis) and phagocytosis of the intruders. It is noteworthy that there is a growing body of evidence that uncontrolled complement biosynthesis and activation in the CNS can contribute to exacerbate the neuronal loss in several neurodegenerative disorders. We provide here an insightful review of the double-edged sword activities of the local innate complement system in the CNS and discuss further the potential therapeutic avenues of delivering complement inhibitors to control brain inflammation. PMID- 11890456 TI - Calcium channels and channelopathies of the central nervous system. AB - Several inherited human neurological disorders can be caused by mutations in genes encoding Ca2+ channel subunits. This review deals with known human and mouse calcium channelopathies of the central nervous system (CNS). The human diseases comprise: 1) a recessive retinal disorder, X-linked congenital stationary night blindness, associated with mutations in the CACNA1F gene, encoding alpha(1)1.4 subunits of L-type channels; and 2) a group of rare allelic autosomal dominant human neurological disorders including familial hemiplegic migraine, episodic ataxia type 2, and spinocerebellar ataxia type 6, all associated with mutations in the CACNA1A gene, encoding alpha(1)2.1 subunits of P/Q-type calcium channels. Mutations at the mouse orthologue of the CACNA1A gene cause a group of recessive neurological disorders, including the tottering, leaner, and rocker phenotypes with ataxia and absence epilepsy, and the rolling Nagoya phenotype with ataxia without seizures. Two other spontaneous mouse mutants with ataxia and absence epilepsy, lethargic and stargazer, have mutations in genes encoding a calcium channel auxiliary beta subunit and a putative calcium channel auxiliary gamma subunit. For each channelopathy, the review describes disease phenotype, channel genotype, and known functional consequences of the pathological mutations; in some cases, it also describes working hypothesis and/or speculations addressing the challenging question of how the alterations in channel function lead to selective cellular dysfunction and disease. PMID- 11890455 TI - Circadian clock system in the pineal gland. AB - The pineal gland is a neuroendocrine organ that functions as a central circadian oscillator in a variety of nonmammalian vertebrates. In many cases, the pineal gland retains photic input and endocrinal-output pathways both linked tightly to the oscillator. This contrasts well with the mammalian pineal gland equipped only with the output of melatonin production that is subject to neuronal regulation by central circadian oscillator located in the suprachiasmatic nucleus (SCN) of the hypothalamus. Molecular studies on animal clock genes were performed first in Drosophila and later developed in rodents. More recently, clock genes such as Per, Cry, Clock, and Bmal have been found in a variety of vertebrate clock structures including the avian pineal gland. The profiles of the temporal change of the clock gene expression in the avian pineal gland are more similar to those in the mammalian SCN rather than to those in the mammalian pineal gland. Avian pineal gland and mammalian SCN seem to share a fundamental molecular framework of the clock oscillator composed of a transcription/translation-based autoregulatory feedback loop. The circadian time-keeping mechanism also requires several post translational events, such as protein translocation and degradation processes, in which protein phosphorylation plays a very important role for the stable 24-h cycling of the oscillator and/or the photic-input pathway for entrainment of the clock. PMID- 11890457 TI - The other half of Hebb: K+ channels and the regulation of neuronal excitability in the hippocampus. AB - Historically, much attention has focused on the mechanisms of activity-dependent plasticity since the description of long-term potentiation by Bliss and Lomo in the early 1970s, while extrasynaptic changes have received much less interest. However, recent work has concentrated on the role of back-propagating action potentials in hippocampal dendrites in synaptic plasticity. In this review, we focus on the modulation of back-propagating action potentials by K+ currents in the dendrites of hippocampal cells. We described the primary K+-channel subunits and their interacting subunits that most likely contribute to these currents, and how these sites can be regulated by phosphorylation and other mechanisms. In conclusion, we provide a model for an alternative form of coincidence detection through K+ channels in the hippocampus. PMID- 11890458 TI - Neuregulin signaling via erbB receptor assemblies in the nervous system. AB - Neuregulins (NRG) play important roles in the development, maintenance, and repair of the nervous system, with influences on neuronal migration, synaptogenesis, receptor subunit composition, and the proliferation/survival of oligodendrocytes and Schwann cells. However, the precise detail of how the NRGs signal through ErbB receptors, particularly at central synapses, is incomplete. The receptor kinase domain provides sites for association with adaptor proteins. In addition, evidence from recent reports suggests that ErbB2/4 receptors, through their C-terminal amino acids, can form specific associations with scaffolding proteins. The existence of such assemblies expands the range of signaling cascades available to the NRGs. PMID- 11890460 TI - The use of quantitative histological and molecular data for risk assessment and biologically based model development. AB - In organs with diverse cell populations, it is not uncommon for one type of cell to respond while others are spared. Even in an organ with common cell types, such as hepatocytes within the liver, the population of cells may respond with different sensitivities for injury or for biochemical responses to toxicants. In the liver, many tumor promoters induce cytochrome P450 enzymes and other proteins in centrilobular cells at much lower doses than required to cause induction in periportal cells. In addition, these induction responses appear to occur at the level of individual cells--a 50% response of the liver for induction does not represent 50% induction in all cells. Instead, half of the cells are fully induced and half are unaffected. Cells "switch" from one phenotypic state to another. Over the past 10 years, several attempts have been made to model these cellular switches and to understand their relevance for hepatic tumor promotion and risk assessment. The data used for analyzing these switches include responses of the entire liver (total induction), responses of individual cells in the liver (regional induction), and cellular responses such as proliferation and apoptosis. This brief overview describes the development of biologically based, dose response (BBDR) models for protein induction and tumor promotion in liver by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with emphasis on the role of specific types of histological and molecular data in providing insights about mechanisms for cellular switches and their implications for tumor promotion. As the biological basis of these switches become unraveled and incorporated into the models, these BBDR models should eventually serve to improve risk assessments with a variety of liver tumor promoters with receptor-based modes of action. PMID- 11890461 TI - Laser technologies in toxicopathology. AB - One of the main concepts in toxicology and risk assessment is the identification of compounds with the least toxicity, gaining increased understanding of the underlying mechanisms of efficacy and toxicity so as to accelerate the early selection of compounds for development. For this purpose, "cutting-edge" technologies, such as flow cytometry (FC), laser scanning cytometry (LSC) and confocal laser scanning microscopy (CLSM), have proved to be valuable tools. FC, LSC and CLSM have been successfully applied in a wide range of areas within toxicology and research including genetics, reproduction, dermatology, pathology and target organ toxicity. The scope of this paper is to give a short overview of the usefulness of the different laser applications. Specific examples of the impact of these technologies will be presented or can be found in the references. Flow cytometry methods have been successfully applied in immunophenotyping, micronuclei scoring, polyploidy determination, apoptosis and cell cycle evaluation, cell proliferation and quantification. A three-parameter FC method for the analysis of testicular toxicity has also been established as an alternative to traditional histopathological methods. This method allows a large number of cells to be analysed in a short time and provides quantitative values to evaluate testicular damage in the rat. Laser scanning cytometry has been used in our unit for rat blood cell immunophenotyping, tumor proliferation, apoptosis and cell cycle analysis on minipig and rat skin and cardiac cells identification. The wide range of applications that can be applied with the LSC shows the enormous potential of this technology in research and development. Confocal laser scanning microscope was used in our laboratory, in collaboration with the research department, to investigate the mechanisms underlying hepatic lesions found in dogs, to detect fibrinogen influx into rat lung, to explore the mechanism of eye toxicity and to quantify dopaminergic fibers in brain sections. Integrating these technologies within discovery pathology allowed us to understand disease processes with respect to their development and subsequent consequences. It contributes to descriptive pathologic diagnostic and allows a productive interaction with research and development. These technologies offer a range of novel applications and have been shown to be useful tools in terms of specificity, sensitivity, reliability, rapidity and quantification. Expertise in cutting-edge technologies, pathology and cell and molecular biology is essential to a successful and flexible interaction across all therapeutic areas in drug discovery. PMID- 11890462 TI - Quantitation of molecular endpoints for the dose-response component of cancer risk assessment. AB - Cancer risk assessment involves the steps of hazard identification, dose-response assessment, exposure assessment, and risk characterization. The rapid advances in the use of molecular biology approaches has had an impact on all 4 components, but the greatest overall current and future impact will be on the dose-response assessment because this requires an understanding of the mechanisms of carcinogenesis, both background and induced by environmental agents. In this regard, hazard identification is a qualitative assessment and dose-response is a quantitative estimate. Thus, the latter will ultimately require a quantitative assessment of molecular endpoints that are used to describe the dose-response for cancer. It has been possible for many years to quantitate alterations at the level of the single gene. For example, analysis of mutation frequency by phenotypic selection, analysis of transcription (mRNA) by Northern blot, analysis of translation (proteins) by Western blot, and analysis of kinetics of metabolism from metabolite levels. However, it is becoming clear that it is necessary when considering risk for adverse health outcomes to develop quantitative approaches for whole cell phenotypes or organ effects. For example, cancer is a whole tissue phenotype, not a feature of single gene mutations, in spite of the multistep (multimutation) mode of formation of a tumor. Thus, there is the need to quantitate the circuitry of a cell: the metabolic/biochemical pathways, genetic regulation pathways, and signaling pathways in normal and stressed conditions. The hypothesis presented by Hanahan and Weinberg of the requirement for 6 acquired characteristics for tumor development, independent of tissue type and species or inducer, seems to provide a viable approach. This hypothesis can be addressed through whole cell molecular assessment using microarrays and quantitative PCR together with the emerging proteomic approaches. This is the world of the new computational cell biology. PMID- 11890463 TI - Current methods for assessing safety of genetically modified crops as exemplified by data on Roundup Ready soybeans. AB - Several laboratories have used recombinant DNA technology in plant breeding to improve compositional, processing, and agronomic characteristics of plants. These transformed plants have been extensively tested in field trials, have gained full regulatory approvals and are currently being marketed in a number of countries around the world. This paper briefly summarizes the approach used to assure the safety of foods and feeds derived from these genetically modified crops, as exemplified by data on Roundup Ready soybeans that has been developed by Monsanto Company using biotechnology in order to confer tolerance to glyphosate, the active ingredient in Roundup herbicide, by the production of the CP4 enolpyruvylshikimate-3-phosphate synthase protein. The results of the studies demonstrate that Roundup Ready soybeans are as safe as traditional soybeans with respect to food and feed safety. PMID- 11890464 TI - Safety assessment and public concerns for genetically modified food products: the Japanese experience. AB - The recombinant DNA (rDNA) technique is expected to bring about great progress in the improvement of breeding technology and the development of new plant varieties showing high quality and high yield, such as those with excellent pest and disease resistance, those with environmental stress tolerance, and so forth. In the United States and Canada, many genetically modified (GM) crop plants were commercialized as early as 1994. In Japan, 35 transgenic crop plants, such as herbicide tolerant soybean, cotton, and canola, and insect-resistant corn, cotton, and potatos, were authorized and considered marketable until April 2001. The general public, however, is not familiar with rDNA technology, and some people seem to feel uncomfortable with biotechnology, frequently because of the difficulty of the technology and lacking of sufficient information. New labeling systems were initiated in April 2001 in Japan to provide information regarding the use of GM crops as raw material. PMID- 11890465 TI - Safety assessments and public concern for genetically modified food products: the American view. AB - In the relatively short time since their commercial introduction in 1996, genetically modified (GM) crops have been rapidly adopted in the United States GM crops are regulated through a coordinated framework developed in 1992 and administered by three agencies-the US Department of Agriculture (USDA) that ensures the products are safe to grow, the Environmental Protection Agency (EPA) that ensures the products are safe for the environment, and the Food and Drug Administration (FDA) that ensures the products are safe to eat. Rigorous food and environmental safety assessments must be completed before GM crops can be commercialized. Fifty-one products have been reviewed by the FDA, including several varieties of corn, soybeans, canola, cotton, rice, sugar beets, potatoes, tomatoes, squash, papaya, and flax. Because FDA considers these crops "substantially equivalent" to their conventional counterparts, no special labeling is required for GM crops in the United States and they are managed as commodities with no segregation or identity preservation. GM crops have thus made their way through commodity distribution channels into thousands of ingredients used in processed foods. It has been estimated that 70% to 85% of processed foods on supermarket shelves in the United States today contain one or more ingredients potentially derived from GM crops. The food industry and retail industry have been monitoring the opinions of their consumers on the GM issue for the past several years. Numerous independent groups have also surveyed consumer concerns about GM foods. The results of these surveys are shared and discussed here. PMID- 11890466 TI - Application of genetically altered models as replacement for the lifetime mouse bioassay in pharmaceutical development. AB - The international pharmaceutical regulatory academic and industrial toxicology communities are collaborating to improve the efficiency and effectiveness of cancer hazard identification based on dramatic improvements in our understanding of the cancer process. Guidelines emanating from the International Conference on Harmonization provide for use of in vivo alternatives. Standard practices utilizing lifetime rat and mouse studies are recognized as seriously flawed with over 80% false positive rates. Furthermore, tobacco, the most important human carcinogen commercialized by industry, is negative in these traditional lifetime studies. The lifetime mouse bioassay is generally recognized in pharmaceutical development as not adding value in safety assessment. An international consortium under the aegis of ILSI has recently completed an evaluation of alternative mouse cancer models. Transgenic models are less expensive, use fewer animals and take less time than traditional lifetime bioassays. These alternative models have now been sufficiently evaluated to be considered useful in the safety assessment plan for pharmaceuticals in development. Specifically for example, the rasH2 appears useful in detecting nongenotoxic as well as genotoxic rodent tumorigens with improved concordance with human response. The p53+/- heterozygous mouse apparently identifies hormonal carcinogenic mechanisms, immunosuppressive carcinogens, and genotoxic carcinogens. The TG:AC predicts for rodent tumorigens applied topically. Recent experiences at FDA, CPMP, and MHW indicate that with good planning and agency interactions, regulatory acceptability can be anticipated. PMID- 11890467 TI - The Tg rasH2 mouse in cancer hazard identification. AB - The Tg rasH2 transgenic mouse has been developed as an altemative to the lifetime mouse bioassay to predict the carcinogenic potential of chemicals. Unlike the p53+/- mouse, the Tg rasH2 mouse is sensitive to both genotoxic and nongenotoxic carcinogens. The Tg rasH2 mouse, officially designated CB6F1-TgN (RasH2), contains multiple copies of the human c-Ha-ras oncogene and promoter within its genome. These mice develop spontaneous andchemically inducedneoplasms earlierin life and in greaternumbersthan wild-type mice, reflectingtheirenhanced sensitivity to neoplasia. The most common spontaneous neoplasms in control Tg rasH2 mice 8 to 9 months of age are lung adenomas and carcinomas (7.4% incidence), splenic hemangiomas and hemangiosarcomas (5.4%), forestomach squamous cell papillomas and carcinomas (2.4%), and skin neoplasms (1.2%). Simulations have demonstrated that 20 to 25 mice/sex/treatment group are required to provide the assay with adequate statistical power. Four of 6 known or suspected human carcinogens tested in Tg rasH2 mice were positive in this assay. For 19 nonmutagenic agents testing positive in conventional rodent bioassays, 7 chemicals were positive, 10 chemicals were negative, and 2 were equivocal. None of the 10 nonmutagenic rodent carcinogens that were negative in the Tg rasH2 mouse model are considered to be human carcinogens. All nonmutagenic chemicals that were negative in the conventional rodent bioassays were also negative in the Tg rasH2 model. Results for 15 of 18 mutagenic chemicals tested in Tg rasH2 mice agreed with the results of conventional rodent bioassays, and 3 results were equivocal. The Tg rasH2 mouse model appears to predict known or suspected human carcinogens as well as the traditional mouse bioassay, but with fewer positive results for nongenotoxic compounds that are not considered human carcinogens. The Tg rasH2 mouse model is the most thoroughly tested in vivo altemative to the lifetime mouse bioassay for nongenotoxic compounds administered by oral or parenteral routes. The U.S. FDA Carcinogenicity Assessment Committee has determined that the Tg rasH2 model has been adequately evaluated for consideration for carcinogenicity testing of pharmaceutical candidates and its use could contribute to the weight of evidence for carcinogenicity assessment. The FDA will consider proposals to replace lifetime mouse carcinogenicity studies with 6-month Tg rasH2 mouse studies to support pharmaceutical registration on a case-by-case basis. PMID- 11890468 TI - The Trp53 hemizygous mouse in pharmaceutical development: points to consider for pathologists. AB - ILSI-HESI sponsored an international consortium for the evaluation of alternative models, including the TrpS3+/- mouse. for use in short-term carcinogenicity testing of pharmaceuticals. Products of the ILSI evaluation included guidance for protocol design and assay interpretation, spontaneous tumor incidences, diagnostic criteria for common proliferative lesions, and results of assays for pharmaceutical agents that are known human and/or rodent carcinogens and non carcinogens. Based on the ILSI evaluation, recommended protocol elements for this model include: 26-week study duration, groups > or = 15/sex/dose, a positive control group (benzene or p-cresidine), a negative control group and 3 dose groups, the high dose set at MTD or MFD, routine in-life evaluations, and complete necropsies with microscopic evaluation of tissues. Favored statistical analyses are trend tests or pair-wise comparisons, with no adjustments for survival. For an assay to be valid, positive control groups must demonstrate an effect, and the MTD or MFD must be reached in both sexes. Criteria for a negative response include a valid assay, no statistical increase in common tumors, no biologically significant numerical increase in rare tumors, and no tumor incidence above that of historical controls. Positive responses can consist of statistically significant increases in the incidence of a common tumor or numerical increases in a rare tumor, which may not be statistically significant. In either case, the incidence should be clearly above historical control values. Evidence of a dose response or occurrence of hyperplasia in a tissue with a neoplastic response can support interpreting an assay as positive. The two most common spontaneous tumors (> 1 %) in Trp53+/- mice are malignant thymic lymphomas and subcutaneous sarcomas. Use of implanted electronic transponders can increase the incidence of sarcomas. Important rare spontaneous tumors (incidence < or = 1%) are osteosarcomas and pulmonary adenomas. Many other tumor types have been reported to occur sporadically in Trp53+/- mice. Diagnostic challenges for this model include differentiating lymphoma from atypical thymic hyperplasia and recognizing the variable histopathology of subcutaneous sarcomas. In reported bioassays, Trp53+/- mice responded positively to genotoxic carcinogens, negatively to non-genotoxic rodent carcinogens, and negatively to noncarcinogens, indicating that unlike the 2-year mouse assay, this short-term assay is not overly sensitive. Positive responses often elicited an increase in tumors that occur spontaneously. To successfully use this model, pathologists must understand the biology of the Trp53 tumor suppressor gene and the principles of protocol design and data interpretation for short-term bioassays. They must also know the historical response pattern of Trp53+/- mice to test agents and be able to accurately diagnose tumors in this model. Use of the Trp53+/- mouse presents the pharmaceutical industry with several challenges, one of which is managing the uncertainty created by a lack of precedents for regulatory decisions about some possible outcomes for short-term carcinogenicity assays. PMID- 11890459 TI - Clustering of nicotinic acetylcholine receptors: from the neuromuscular junction to interneuronal synapses. AB - Fast and accurate synaptic transmission requires high-density accumulation of neurotransmitter receptors in the postsynaptic membrane. During development of the neuromuscular junction, clustering of acetylcholine receptors (AChR) is one of the first signs of postsynaptic specialization and is induced by nerve released agrin. Recent studies have revealed that different mechanisms regulate assembly vs stabilization of AChR clusters and of the postsynaptic apparatus. MuSK, a receptor tyrosine kinase and component of the agrin receptor, and rapsyn, an AChR-associated anchoring protein, play crucial roles in the postsynaptic assembly. Once formed, AChR clusters and the postsynaptic membrane are stabilized by components of the dystrophin/utrophin glycoprotein complex, some of which also direct aspects of synaptic maturation such as formation of postjunctional folds. Nicotinic receptors are also expressed across the peripheral and central nervous system (PNS/CNS). These receptors are localized not only at the pre- but also at the postsynaptic sites where they carry out major synaptic transmission. In neurons, they are found as clusters at synaptic or extrasynaptic sites, suggesting that different mechanisms might underlie this specific localization of nicotinic receptors. This review summarizes the current knowledge about formation and stabilization of the postsynaptic apparatus at the neuromuscular junction and extends this to explore the synaptic structures of interneuronal cholinergic synapses. PMID- 11890469 TI - Toxicogenomics, drug discovery, and the pathologist. AB - The field of toxicogenomics, which currently focuses on the application of large scale differential gene expression (DGE) data to toxicology, is starting to influence drug discovery and development in the pharmaceutical industry. Toxicological pathologists, who play key roles in the development of therapeutic agents, have much to contribute to DGE studies, especially in the experimental design and interpretation phases. The intelligent application of DGE to drug discovery can reveal the potential for both desired (therapeutic) and undesired (toxic) responses. The pathologist's understanding of anatomic, physiologic, biochemical, immune, and other underlying factors that drive mechanisms of tissue responses to noxious agents turns a bewildering array of gene expression data into focused research programs. The latter process is critical for the successful application of DGE to toxicology. Pattern recognition is a useful first step, but mechanistically based DGE interpretation is where the long-term future of these new technologies lies. Pathologists trained to carry out such interpretations will become important members of the research teams needed to successfully apply these technologies to drug discovery and safety assessment. As a pathologist using DGE, you will need to learn to read DGE data in the same way you learned to read glass slides, patiently and with a desire to learn and, later, to teach. In return, you will gain a greater depth of understanding of cell and tissue function, both in health and disease. PMID- 11890470 TI - Alternatives models in carcinogenicity testing--a European perspective. AB - Transgenic mouse strains offer the prospect of significant benefits in the in vivo assessment of carcinogenic potential. The European Regulatory Authorities have been supportive of their inclusion as one of the second-test options in the International Conference on Harmonization of Technical Requirements for the Registration of Pharmaceuticals for Human use (ICH). However, there is a concern regarding premature systematic use of these models. At present, the information from the International Life Sciences Institute (ILSI) project suggests that the transgenic models under study are similarly sensitive to genotoxic pharmaceuticals. There are apparently some false negatives and false positives. For regulatory purposes, it is not yet possible to differentiate the models with respect to hazard identification and risk assessment. The evaluation of the models has reached an interesting but, at certain points, equivocal stage. Based on the weight of evidence gathered thus far, regulatory authorities cannot neglect the outcome of such studies but need to be cautious in their interpretation of data from such models, and the application in risk assessment procedures. PMID- 11890471 TI - Toxicologic pathology in the new millennium. PMID- 11890472 TI - Introduction and summary of the workshop "Drug Research Support: Selection and Early Development". PMID- 11890473 TI - SK&F 95654-induced acute cardiovascular toxicity in Sprague-Dawley rats- histopathologic, electron microscopic, and immunohistochemical studies. AB - The characteristics and pathogenesis of the cardiovascular toxicity induced by the type III selective phosphodiesterase inhibitor SK&F 95654 were examined in 2 studies. Sprague-Dawley rats received either a single sc injection of 50, 100, or 200 mg/kg SK&F 95654 and were euthanized at 24 hours after administration of the drug (Study 1), or were given a single subcutaneous (sc) injection of 100 mg/kg SK&F 95654 and euthanized at 1, 2, 4, 6, 8,12, 24 hours, or 2 weeks after treatment (Study 2). Control rats received either DMSO or saline. Myocardial lesions and vascular lesions of the mesentery, spleen, and pancreas were seen 24 hours after dosing with either 50,100, or 200 mg/kg SK&F 95654. The frequency and severity of these lesions (evaluated after the 100 mg/kg dose) increased with time over a period of 1 to 24 hours. By 2 weeks, the lesions subsided. Cardiac lesions consisted of myocyte necrosis with hypercontraction bands, inflammatory cell infiltration, interstitial hemorrhage, and interstitial edema. Vascular lesions of the mesentery were most prominent and consisted of vasodilatation and inflammation in the small-sized vessels, arterial medial necrosis and hemorrhage, and venous thrombosis. The vascular lesions included: leukocyte adhesion to endothelial cells, transendothelial migration of leukocytes, and inflammatory cell infiltration into vessel walls. Affected vessels included arteries, terminal arterioles, capillaries, postcapillary venules, and veins. Apoptosis of endothelial and smooth muscle cells was detected in the mesenteric vasculature by both TUNEL assay and electron microscopy. Evidence of endothelial cell activation in the mesenteric arteries and veins was also observed by electron microscopy. Immunohistochemical staining detected enhanced endothelial cell expression of intercellular adhesion molecule- 1 (ICAM- 1) and von Willebrand factor (vWF) in the mesenteric arteries and veins. Mast cells were noted to be more prevalent in affected mesenteric tissue from drug-treated animals. The present findings suggest that apoptosis of endothelial and smooth muscle cells, activation of endothelial cells, recruitment of mast cells, and increased expression of adhesion molecules are important factors to the overall pathogenesis of SK&F 95654-induced vasculitis. PMID- 11890474 TI - Strategic importance of research support through pathology. AB - The pace at which new drug candidates are being identified by Discovery Research demands that they be screened for preclinical attributes rapidly and efficiently. The early identification and elimination of compounds with toxic liabilities will produce safer drugs in a shorter time period, and with an increased rate of success. Most major pharmaceutical companies now recognize the strategic role of pathology support for research and have developed specific units to effect this outcome. The early interaction of these pathologists with drug discovery teams to identify compounds with toxic liabilities is critical. Approaches being used include high throughput in vitro screens to predict the relative toxicity of discovery compounds and to provide early indications of underlying mechanisms, target profiling to predict consequences of receptor-ligand interactions at other than-indicated target sites, and acute in vivo studies to establish tolerability limits and target organs of toxicity. These approaches include the application of contemporary tools such as genomics, proteomics, metabonomics, and genetically engineered animal models. To maximize the benefit of discovery pathology, it is critical that pharmaceutical companies also actively participate in non proprietary knowledge sharing and the education of pathologists and toxicologists to lead these efforts in the future. PMID- 11890475 TI - Alteration of liver cell function and proliferation: differentiation between adaptation and toxicity. AB - Exposure of experimental animals to biologically effective levels of chemicals, either endogenous or exogenous, the latter of either synthetic or natural origin, elicits a response(s) that reflects the diverse ways in which the various units of organization of an organism deal with chemical perturbation. For some chemicals, an initial response constitutes an adaptive effect that maintains homeostasis. Disruption of this equilibrium at any level of organization leads to an adverse effect, or toxicity. The livers of laboratory animals and humans, like other organs, undergo programmed phases of growth and development, characterized by proliferation followed by differentiation. With organ maturity, the process of differentiation leads to the commitment of differentiated cells to constitutive functions that maintain homeostasis and to specialized functions that serve organismal needs. In the mature livers of all species, proliferation of all cell types subsides to a low level, Thus, the mature liver consists of 2 types of cells: intermediate cells, the hepatocytes, which replicate infrequently, but can respond to signals for replication, and replicating cells, the stem cells, endothelial, Kupffer, and stellate cells (Ito or pericytes), bile duct epithelium, and granular lymphocytes (pit cells). Quantifiable alterations or effects at the molecular level underlie alterations at the organelle level, which in turn lead to alterations at the cellular level, which can ultimately be manifested as a change in the whole organism. Alterations can be quantal (binary), either all or none, as with cell replication, cell necrosis or apoptosis, and cell differentiation, which take place at the cellular level. They can also be graded or continuous (nonbinary), as with enzyme induction, organelle hypertrophy, and extracellular matrix elaboration, occurring either at the intra- or extra (supra) cellular level. Any quantifiable change induced in the function or structure of a cell or tissue constitutes a response or effect. Each of the several types of cell in the liver responds to a given stimulus according to its localization and function. Generally, renewing cells are more vulnerable to chemical injury than intermediate cells, which are largely quiescent. Hepatic adaptive responses usually involve actions of the chemical on cellular regulatory pathways, often receptor mediated, leading to changes in gene expression and ultimately alteration of the metabolome. The response is directed toward maintaining homeostasis through modulation of various cellular and extracellular functions. At all levels of organization, adaptive responses are beneficial in that they enhance the capacity of all units to respond to chemical induced stress, are reversible and preserve viability. Such adaptation at subtoxic exposures is also referred to as hormesis. In contrast, adverse or toxic effects in the liver often involve chemical reaction with cellular macromolecules and produce disruption of homeostasis. Such effects diminish the capacity for response, can be nonreversible at all levels of organization, and can compromise viability. An exposure that elicits an adaptive response can produce toxicity with longer or higher exposures (ie, above a threshold) and the mechanism of action changes with the effective dose. A variety of hepatic adaptive and toxic effects has been identified. Examples of adaptive effects are provided by phenobarbital and ciprofibrate, whereas p-dichlorobenzene and 2 acetylaminofluorene illustrate different toxic effects. The effects of chemicals in the liver are, in general, similar between experimental animals and humans, although exceptions exist. Thus, identification and monitoring of both types of effect are integral in the safety assessment of chemical exposures. PMID- 11890476 TI - Immune responses: adverse versus non-adverse effects. AB - The adaptive immune system in vertebrates has evolved to provide host resistance to infectious microorganisms and malignant disease. Normal immune function and the induction of specific immune responses require the orchestrated interaction between cells and molecules both within and outside the lymphoid system. Immunotoxicology can be defined as the study of adverse health effects that may result from the interaction of xenobiotics with the immune system. In general terms such effects can take one of two forms. The first of these is immunotoxicity (or immunosuppression) where there is a perturbation of, or damage to, one or more components of the immune system resulting in impaired immune function and reduced host resistance. The design and interpretation of experimental immunotoxicity studies and the investigation of clinical immunosuppression require consideration of the relationship between changes in the structure and/or function of discrete components of the immune system and holistic changes in the susceptibility to infectious and malignant disease. The other main way in which chemicals may cause adverse health effects secondary to interaction with the immune system is through stimulation of specific immune responses that result in allergic disease. Allergy to chemicals and proteins can take many forms, including allergic contact dermatitis, allergic sensitization of the respiratory tract (associated with rhinitis and/or asthma), systemic allergic reactions (associated frequently with drug treatment), and gastrointestinal disease. Here there is a need to distinguish between immunogenic responses per se and those immune responses that are of sufficient vigor and of the quality necessary to provoke allergic sensitization. The purpose of this article is to explore the extent to which distinctions can be drawn between adverse and nonadverse effects in the context of immunotoxicity and allergy. PMID- 11890477 TI - Recognition of adverse and nonadverse effects in toxicity studies. AB - One of the most important quantitative outputs from toxicity studies is identification of the highest exposure level (dose or concentration) that does not cause treatment related effects that could be considered relevant to human health risk assessment. A review of regulatory and other scientific literature and of current practices has revealed a lack of consistency in definition and application of frequently used terms such as No Observed Effect Level (NOEL), No Observed Adverse Effect Level (NOAEL), adverse effect, biologically significant effect, or toxicologically significant effect. Moreover, no coherent criteria were found that could be used to guide consistent interpretation of toxicity studies, including the recognition and differentiation between adverse and nonadverse effects. This presentation will address these issues identified first by proposing a standard set of definitions for key terms such as NOEL and NOAEL that are frequently used to describe the overall outcome of a toxicity study. Second, a coherent framework is outlined that can assist the toxicologist in arriving at consistent study interpretation. This structured process involves two main steps. In the first, the toxicologist must decide whether differences from control values are treatment related or if they are chance deviations. In the second step, only those differences judged to be effects are further evaluated in order to discriminate between those that are adverse and those that are not. For each step, criteria are described that can be used to make consistent judgments. In differentiating an effect from a chance finding, consideration is given inter alia to dose response, spurious measurements in individual parameters, the precision of the measurement under evaluation, ranges of natural variation and the overall biological plausibility of the observation. In discriminating between the adverse and the non-adverse effect consideration is given to: whether the effect is an adaptive response, whether it is transient, the magnitude of the effect, its association with effects in other related endpoints, whether it is a precursor to a more significant effect, whether it has an effect on the overall function of the organism. whether it is a specific effect on an organ or organ system or secondary to general toxicity or whether the effect is a predictable consequence of the experimental model. In interpreting complex studies it is recognised that a weight of the evidence approach, combining the criteria outlined here to reach an overall judgment, is the optimal way of applying the process. It is believed that the use of such a scheme will help to improve the consistency of study interpretation that is the foundation of hazard and risk assessment. PMID- 11890478 TI - The north american control animal database: a resource based on standardized nomenclature and diagnostic criteria. AB - Historical control data have been shown to be valuable in the interpretation and evaluation of results from rodent carcinogenicity studies. Standardization of terminology and histopathology procedures is a prerequisite for meaningful comparison of control data across studies and analysis of potential carcinogenic effects. Standardization is particularly critical for the construction of a database that includes incidence data from different studies evaluated by pathologists in different laboratories. Standardized nomenclature and diagnostic criteria have been established for neoplasms and proliferative lesions. Efforts of the National Toxicology Program, the Society of Toxicologic Pathology (STP), and the Registry of Industrial Toxicology Animal-data (RITA) have led to a harmonized pathology nomenclature for the rat and the mouse. This nomenclature with detailed descriptions of lesions is available in publications by the STP and International Agency for Research on Cancer (IARC). A listing of these terms is available on the World Wide Web. Utilizing the model established by RITA and working with the International Life Sciences Institute (ILSI), companies with laboratories in North America formed a working group in 1994 to establish and maintain a database of neoplastic and proliferative lesions from control animals in carcinogenicity studies. The rationale for development of the North American Control Animal Database (NACAD), the factors that influence tumor incidence, operation of the database, and the benefits to be realized by using a standardized approach are discussed. PMID- 11890480 TI - The value of historical control data-scientific advantages for pathologists, industry and agencies. AB - Historical control tumor data are useful in the interpretation of long-term rodent carcinogenicity bioassays, especially to assess the occurrence of rare tumors and marginally increased tumor incidences. The major prerequisites to compare historical control data with studies under evaluation are the validity and consistency of the respective databases. The RITA (Registry of Industrial Toxicology Animal-data) database for historical data of tumors and pre-neoplastic lesions collects data according to highly standardized procedures including tissue sampling and trimming, histopathology according to internationally harmonized nomenclature and diagnostic criteria, and peer review. All lesions that are entered are unanimously diagnosed according to IARC (Intermational Agency for Research on Cancer)/WHO criteria. The validity of data is additionally confirmed by a complete peer review performed by a database pathologist. Equivocal diagnoses and selected cases are additionally submitted to a panel of RITA pathologists. In the RITA database, there are currently 10,896 rats from 106 studies with more than 17,604 primary tumors and 16,551 pre-neoplastic lesions. The RITA database for historical control data for Wistar and Sprague Dawley rats as well as for different mouse strains is briefly described. Based upon RITA background data, the survival rate of Wistar rats has been consistent over a period of 10 years. The occurrence of tumor-bearing animals also shows a stable percentage over a decade. Additionally, examples of how historical control data may support carcinogenic risk assessment in cases of rare tumors or marginally increased incidences of tumors and pre-neoplastic lesions are given. PMID- 11890479 TI - Practical aspects of discovery pathology. AB - Pathologists are uniquely qualified to play a central role in driving drug discovery and development programs by: 1) establishing disease models to assess potential therapies, 2) characterizing modifications in the disease state in response to therapies, 3) characterizing toxicologic mechanisms and responses to drug candidates, and 4) facilitating multidisciplinary efforts to monitor for the clinical occurrence, progression, and reversibility of adverse events. Such nontraditional deployment of resources must, to be viable, produce benefits to the pharmaceutical industry comparable to those of more conventional activities such as delivery of data in nonclinical safety studies. Additionally, benefits must be tangible from standpoints such as time savings or improved quality of research decisions, manifesting as either program acceleration or improved candidate survival. PMID- 11890481 TI - Quality review procedures necessary for rodent pathology databases and toxicogenomic studies: the National Toxicology Program experience. AB - Accuracy of the pathology data is crucial since rodent studies often provide critical data used for setting human chemical exposure standards. Diagnoses represent a judgment on the expected biological behavior of a lesion and peer review can improve diagnostic accuracy and consistency. With the conduct of 500 2 year rodent studies, the National Toxicology Program (NTP) has refined its process for comprehensive review of the pathology data and diagnoses. We have found that careful judgment can improve and simplify the review, whereas simply applying a set review procedure may not assure study quality. The use of reviewing pathologists and pathology peer review groups is a very effective procedure to increase study quality with minimal time and cost. New genomic technology to assess differential gene expression is being used to predict morphological phenotypes such as necrosis, hyperplasia, and neoplasia. The challenge for pathologists is to provide uniform pathology phenotypes that can be correlated with the gene expression changes. The lessons learned in assuring data quality in standard rodent studies also applies to the emerging field of toxicogenomics. PMID- 11890482 TI - Qualitative and quantitative analysis of nonneoplastic lesions in toxicology studies. AB - A pathology report is written to convey information concerning the pathologic findings in a study. This type of report must be complete, accurate and communicate the relative importance of various findings in a study. The overall quality of the report is determined by three Quality Indicators: thoroughness, accuracy, and consistency. Thoroughness is the identification of every lesion present in a particular organ or tissue, including spontaneous background lesions. Experienced pathologists familiar with background lesions may disregard certain types of lesions or establish a threshold or a severity above which background lesions are diagnosed. Accuracy is the ability to make, and precisely communicate, correct diagnoses. Nomenclature of lesions is a matter of definition and experienced pathologists generally agree as to what terms are to be used. Consistency is the uniform use of a specific term to record a defined lesion and implies that the same diagnostic criteria are being followed for each type of diagnosis. The relative severity of nonneoplastic lesions can be recorded either semiquantitatively or quantitatively. Semiquantitative analysis involves the application of defined severity grades or ranges for specific lesions. Quantitative analysis (counts and measurements) can be performed manually or electronically, utilizing image analysis and stereological techniques to provide numerical values. When both qualitative and quantitative parameters are applied in preparation of a pathology report, the recorded pathology findings can be interpreted and put into perspective. The use of this approach assures a reader that the pathology report meets the highest standards. PMID- 11890483 TI - Quantitation of the cancer process in C57BL/6J, B6C3F1 and C3H/HeJ mice. AB - This study examines growth alterations in liver foci and tumor development as a basis for the different susceptibility in hepatocarcinogenesis found among different strains of mice. Male C57, B6C3F1, and C3H mice treated with a single dose (1 mg/kg) of N,N-diethylnitrosamine (DEN) at 15 days of age and followed up to 12 months displayed a strain-dependent (C3H > B6C3F1 > C57) increase in incidence, number, volume fraction, and size of foci and macroscopic lesions (masses). DEN-treated mice exhibited a time-dependent increase in foci size but not in foci number. Phenobarbital (PB) treatment (500 ppm) in the drinking water starting 2 weeks after DEN-initiation did not affect the incidence or number of masses and foci. In all 3 strains, the bromodeoxyuridine labeling index in foci correlated with foci growth, supporting the major role of cell proliferation in foci growth. Measurements of apoptosis by morphological criteria with H&E staining suggest that intrafocal apoptosis may be a late event preventing foci growth and possibly also promoting focal cell selection, whereas extrafocal apoptosis may facilitate clonal growth by removing adjacent normal cells. The onset of conversion of foci to masses also correlated with strain susceptibility to hepatocarcinogenesis. PMID- 11890484 TI - Rapid emergency department intervention for older people reduces risk of functional decline: results of a multicenter randomized trial. AB - OBJECTIVES: To determine the effectiveness of a two-stage (screening and nursing assessment) intervention for older patients in the emergency department (ED) who are at increased risk of functional decline and other adverse outcomes. DESIGN: Controlled trial, randomized by day of ED visit, with follow-up at 1 and 4 months. SETTING: Four university-affiliated hospitals in Montreal. PARTICIPANTS: Patients age 65 and older expected to be released from the ED to the community with a score of 2 or more on the Identification of Seniors At Risk (ISAR) screening tool and their primary family caregivers. One hundred seventy-eight were randomized to the intervention, 210 to usual care. INTERVENTION: The intervention consisted of disclosure of results of the ISAR screen, a brief standardized nursing assessment in the ED, notification of the primary care physician and home care providers, and other referrals as needed. The control group received usual care, without disclosure of the screening result. MEASUREMENTS: Patient outcomes assessed at 4 months after enrollment included functional decline (increased dependence on the Older American Resources and Services activities of daily living scale or death) and depressive symptoms (as assessed by the short Geriatric Depression Scale). Caregiver outcomes, also assessed at baseline and 4 months, included the physical and mental summary scales of the Medical Outcomes Study Short Form-36. Patient and caregiver satisfaction with care were assessed 1 month after enrollment. RESULTS: The intervention increased the rate of referral to the primary care physician and to home care services. The intervention was associated with a significantly reduced rate of functional decline at 4 months, in both unadjusted (odds ratio (OR) = 0.60, 95% confidence interval (CI) = 0.36-0.99) and adjusted (OR = 0.53, 95% CI = 0.31-0.91) analyses. There was no intervention effect on patient depressive symptoms, caregiver outcomes, or satisfaction with care. CONCLUSION: A two-stage ED intervention, consisting of screening with the ISAR tool followed by a brief, standardized nursing assessment and referral to primary and home care services, significantly reduced the rate of subsequent functional decline. PMID- 11890485 TI - Effects of supporting community-living demented patients and their caregivers: a randomized trial. AB - OBJECTIVES: To determine whether community care of demented patients can be prolonged by means of a 2-year support program based on nurse case management. DESIGN: Randomized controlled intervention study with 2-year follow-up. SETTING: Demented patients entitled to payments from the Social Insurance Institution for community care, in five municipalities in eastern Finland. PARTICIPANTS: One hundred demented patients, age 65 and older, living at home with the primary support of informal caregivers, allocated at random to the intervention (n = 53) or control group (n = 47). INTERVENTION: Intervention patients and their caregivers were provided with a 2-year intervention program of systematic, comprehensive support by a dementia family care coordinator. MEASUREMENTS: Time to institutionalization (period in community care) from enrollment of patients in the study to their placement in long-term institutional care. RESULTS: During the first months, the rate of institutionalization was significantly lower in the intervention group than in the control group (P = .042), but the benefit of the intervention decreased with time (P = .028). Estimated probability of staying in community care up to 6, 12, and 24 months was 0.98, 0.92, and 0.63 in the intervention group and 0.91, 0.81, and 0.68 in the control group, respectively. Results also suggest that the intervention used in the study might be especially beneficial to patients with severe dementia and those with problems threatening the continuity of community care. CONCLUSIONS: The placement of demented patients in long-term institutional care can be deferred with the support of a dementia family care coordinator. However, by the end of the 2-year intervention, the number of patients institutionalized was similar in the intervention and control group. It seems to be beneficial to direct this type of intensive support at severely demented patients and their caregivers. On the basis of our experiences, we suggest that intervention by a dementia family care coordinator should be targeted especially at patients with problems threatening the continuity of community care. PMID- 11890486 TI - Impact of a new assessment system, the MDS-HC, on function and hospitalization of homebound older people: a controlled clinical trial. AB - OBJECTIVES: To evaluate the impact of a new assessment system, the Minimum Data Set for Home Care (MDS-HC), on the functional status and hospitalization rates of frail, community-dwelling older people. DESIGN: Single-blind randomized trial with 1-year follow-up. SETTING: Bergamo, Italy. PARTICIPANTS: All 187 subjects who were eligible for home care services delivered by two Health Districts between September 1998 and April 1999. INTERVENTION: Random allocation to an intervention group undergoing MDS-HC assessment or to a control group receiving conventional geriatric assessment with Barthel, Lawton and Brody, and Mini-Mental State Examination (MMSE) scales. MEASUREMENTS: Hospitalization, health services use and costs, and variations in functional status. RESULTS: Survival analysis indicated that the intervention group was admitted to the hospital later and less often than were controls (relative risk = 0.49, 95% confidence interval = 0.56 0.97). Health services were used to the same extent, but intervention subjects used more in-home help services. Total costs for the intervention group were 21% lower than for the control group. The adjusted mean scores of the activities of daily living index (51.7+/-36.1 vs 46.3+/-33.7; P = .05) and MMSE (19.9+/-8.9 vs 19.2+/-10.7; P = .03) were significantly improved in the intervention group as compared with the control group. CONCLUSIONS: The MDS-HC assessment instrument may provide a cost-saving approach to reducing institutionalization and functional decline in older people living in the community. PMID- 11890487 TI - Does behavioral improvement with haloperidol or trazodone treatment depend on psychosis or mood symptoms in patients with dementia? AB - OBJECTIVES: Several previous studies have examined the effects of pharmacological interventions for agitated behavior in patients with dementia. However, the choice of medication in clinical practice continues to be directed largely by local pharmacotherapy culture rather than empirical treatment guidelines. We examined the relationship between behavioral improvement and co-occurring delusions and mood symptoms in patients with dementia who were treated with haloperidol, an antipsychotic medication, or trazodone, a serotonergic antidepressant. DESIGN: Randomized, double-blind, parallel-group, 9-week treatment trial. SETTING: Inpatient geropsychiatry unit. PARTICIPANTS: Twenty eight patients with dementia and agitated or aggressive behaviors. INTERVENTION: Haloperidol 1 to 5 mg/day or trazodone 50 to 250 mg/day. MEASUREMENTS: Cohen Mansfield Agitation Inventory (CMAI), Hamilton Depression Rating Scale (Ham-D), and delusional thoughts subscale and hallucinations subscale of the Behavioral Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD). RESULTS: CMAI scores improved in each treatment group over the 9 weeks of treatment (P < .001 in each group). Within the haloperidol treatment group, CMAI improvement was not associated with baseline delusional thoughts score or with change in delusional thoughts score over the course of treatment. Within the trazodone treatment group, CMAI improvement was associated with baseline score on total Ham-D (r = 0.60, P = .02), Ham-D items measuring subjective mood symptoms (r = -0.50, P = .07), and Ham-D items measuring neurovegetative signs (r = -0.49, P = .08). CMAI improvement was also associated with improvement in Ham-D total score over the course of treatment (r = 0.62, P = .02). CONCLUSIONS: Mild depressive symptoms in patients with dementia and agitated behavior are associated with greater behavioral improvement by trazodone-treated patients. In contrast, the presence of delusions in concert with behavioral disturbance does not necessarily predict greater behavioral improvement with haloperidol treatment than in subjects without signs of psychosis. PMID- 11890488 TI - Frequency of Alzheimer's disease and other dementias in a community outreach sample of Hispanics. AB - OBJECTIVES: To determine the proportion of Alzheimer's disease (AD) and other dementia types in a community sample of Hispanics. DESIGN: This is a descriptive diagnostic study of a nonrandom community outreach sample utilizing established criteria for the diagnosis of dementia type. Recruitment involved direct community outreach with diagnostic evaluations conducted at a university affiliated outpatient clinic. SETTING: Hispanic Neuropsychiatric and Memory Research Clinic at the Olive View-UCLA Medical Center in Sylmar, California. PARTICIPANTS: One hundred community-dwelling Hispanics age 55 and older without prior diagnosis or treatment of their cognitive symptoms. MEASUREMENTS: Each subject underwent a complete medical diagnostic evaluation, in Spanish, including neuropsychological tests, neurological examination, laboratory tests, and brain imaging (computed tomography or magnetic resonance imaging) to establish dementia type. Presence of dementia was established according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria. Diagnosis for probable or possible AD and vascular dementia (VascD) was established using criteria from the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association for probable AD and by research criteria from the National Institute of Neurological Disorders and Stroke and the Association Internationale pour la Recherche et l'Enseignement en Neurosciences for VascD, respectively. Frontotemporal dementia was diagnosed using recommendations set forth by the Lund and Manchester groups. RESULTS: Subjects were poor, with low acculturation levels despite long years of U.S. residence. Forty percent of subjects had had undiagnosed cognitive symptoms for 3 or more years. Of those demented, 38.5% had AD and 38.5% met criteria for VascD. The best predictors of VascD were hypertension and cerebrovascular disease, whereas apolipoprotein E4 allele best predicted AD. Other forms of dementia were also present. Twenty percent of the sample was clinically depressed but not demented. CONCLUSIONS: In comparison with data from predominantly white populations, our proportion of AD cases was lower and that of VascD cases was considerably higher than anticipated. The percentage of clinically depressed older individuals was also high. These findings could have implications for differential cultural and genetic risk factors for dementia among diverse ethnic/racial groups. Further studies are needed to obtain accurate prevalence estimates of dementing disorders among the different U.S. Hispanic populations. PMID- 11890489 TI - Weight change in old age and its association with mortality. AB - OBJECTIVES: Previous studies of weight change and mortality in older adults have relied on self-reported weight loss, have not evaluated weight gain, or have had limited information on health status. Our objective was to determine whether 5% weight gain or loss in 3 years was predictive of mortality in a large sample of older adults. DESIGN: Longitudinal observational cohort study. SETTING: Four U.S. communities. PARTICIPANTS: Four thousand seven hundred fourteen community dwelling older adults, age 65 and older. MEASUREMENTS: Weight gain or loss of 5% in a 3-year period was examined in relationship to baseline health status and interim health events. Risk for subsequent mortality was estimated in those with weight loss or weight gain compared with the group whose weight was stable. RESULTS: Weight changes occurred in 34.6% of women and 27.3% of men, with weight loss being more frequent than gain. Weight loss was associated with older age, black race, higher weight, lower waist circumference, current smoking, stroke, any hospitalization, death of a spouse, activities of daily living disability, lower grip strength, and slower gait speed. Weight loss but not weight gain of 5% or more was associated with an increased risk of mortality that persisted after multivariate adjustment (Hazard ratio (HR) = 1.67, 95% CI = 1.29-2.15) and was similar in those with no serious illness in the period of weight change. Those with weight loss and low baseline weight had the highest crude mortality rate, although the HR for weight loss was similar for all tertiles of baseline weight and for those with or without a special diet, compared with those whose weight was stable. CONCLUSIONS: This study confirms that even modest decline in body weight is an important and independent marker of risk of mortality in older adults. PMID- 11890490 TI - The effect of middle- and old-age body mass index on short-term mortality in older people. AB - OBJECTIVES: To determine the effect of body mass index (BMI) at old age and at age 50 on short-term survival among persons age 65 and older. DESIGN: Cross sectional, using the 4,791 respondents to the community interview of the 1994 National Long Term Care Survey (NLTCS). SETTING: United States of America. PARTICIPANTS: Persons age 65 and older who lived in community settings as of the 1994 NLTCS interview. MEASUREMENTS: Short-term mortality was measured from the date of the 1994 NLTCS through year-end 1995. BMI (kg/m2) (at three points: 1994 NLTCS, 1 year before, age 50) and all other variables, including three other modifiable risk factors known to be related to mortality--cigarette smoking, alcohol consumption, and exercise--were based on self-report. RESULTS: Both the unadjusted and adjusted nadirs of mortality in relation to BMI at old age were found in older persons with a BMI between 30 and 34.9; this was true for males and females in all age groups. The highest mortality rates were found for older persons with very low BMI (<18.5). In contrast, BMI at age 50 was positively related to mortality, with those in the lowest BMI category (<18.5) at age 50 having the lowest mortality. Persons who were obese at age 50 and who were no longer obese at the 1994 NLTCS had lower mortality than persons with stable weight. CONCLUSIONS: Weight reduction by middle-aged persons who are obese should be reinforced as a public health priority, because there is evidence that long term weight loss results in better short-term survival. Further study of healthy older survivors to determine why they are not harmed by heavier weight in old age may provide useful insights into successful aging. PMID- 11890491 TI - Environmental risk factors for delirium in hospitalized older people. AB - OBJECTIVES: To evaluate the relationship of environmental risk factors in hospitals to changes over time in delirium symptom severity scores. DESIGN: Observational prospective clinical study with repeated measurements, several times during the first week of hospitalization and then weekly during hospitalization. SETTING: University-affiliated general community hospital. PARTICIPANTS: Four hundred forty-four patients age 65 and older admitted to the medical wards: 326 with delirium and 118 without delirium. Patients with prior cognitive impairment were oversampled. MEASUREMENTS: The severity of delirium symptoms was measured with the Delirium Index, a scale developed and validated by our group, based on the Confusion Assessment Method. Potential environmental risk factors assessed included isolation, hospital unit, room changes, levels of sensory stimulation, aids to orientation, and presence of medical (e.g., intravenous) or physical restraints. RESULTS: Controlling for initial severity of delirium and patient characteristics, variables significantly related to an increase in delirium severity scores included hospital unit (intensive care or long-term care unit), number of room changes, absence of a clock or watch, absence of reading glasses, presence of a family member, and presence of medical or physical restraints. CONCLUSION: The associations of intensive care and medical and physical restraints with severity of delirium symptoms may be due to uncontrolled confounding by indication. However, the other factors identified suggest potentially modifiable risk factors for symptoms of delirium in hospitalized older people. PMID- 11890492 TI - Delirium before and after operation for femoral neck fracture. AB - OBJECTIVES: The aim of this study was to investigate the differences between preoperative and postoperative delirium regarding predisposing, precipitating factors and outcome in older patients admitted to hospital with femoral neck fractures. DESIGN: A prospective clinical assessment of patients treated for femoral neck fractures. SETTING: Department of orthopedic surgery at Umea University Hospital, Sweden. PARTICIPANTS: One hundred one patients, age 65 and older admitted to the hospital for treatment of femoral neck fractures. MEASUREMENTS: The Organic Brain Syndrome (OBS) Scale. RESULTS: Thirty patients (29.7%) were delirious before surgery and another 19 (18.8%) developed delirium postoperatively. Of those who were delirious preoperatively, all but one remained delirious postoperatively. The majority of those delirious before surgery were demented, had been treated with drugs with anticholinergic properties (mainly neuroleptics), had had previous episodes of delirium, and had fallen indoors. Patients who developed postoperative delirium had perioperative falls in blood pressure and had more postoperative complications such as infections. Male patients were more often delirious both preoperatively and postoperatively. Patients with preoperative delirium were more often discharged to institutional care and had poorer walking ability both on discharge and after 6 months than did patients with postoperative delirium only. CONCLUSIONS: Because preoperative and postoperative delirium are associated with different risk factors it is necessary to devise different strategies for their prevention. PMID- 11890493 TI - Progressive trends in the prevalence of benzodiazepine prescribing in older people in Ontario, Canada. AB - OBJECTIVES: The extensive use of benzodiazepines has been a concern of healthcare providers and policy makers in Canada and around the world. The purpose of this study was to examine temporal trends in benzodiazepine prescriptions dispensed in older people from 1993-1998. DESIGN: Retrospective population-based cross sectional study using administrative databases. SETTING: Ontario, Canada. PARTICIPANTS: The over 1 million residents of Ontario age 65 and older covered by the provincial universal drug benefit program. MEASUREMENTS: The main outcome measures were the prevalence, overall and with respect to age and gender, of benzodiazepine prescriptions dispensed and the ratio of the number of people to whom short- versus long-acting benzodiazepine prescriptions were dispensed in each study year. The annual rates of switching to other psychotropic agents were examined for those patients that discontinued filling benzodiazepine prescriptions. RESULTS: The annual prevalence of benzodiazepine prescriptions dispensed decreased consistently over time (25.1% in 1993 to 22.5% in 1998; P < .001). Benzodiazepine dispensing prevalence increased with increasing age (approximately 20% of those age 65 to 69 to approximately 30% of those age > or =85; P < .001) and more females than males received the medication (relative risk = 1.50, 95% confidence interval = 1.49-1.51). The ratio of short- to long-acting benzodiazepine prescriptions filled increased over time (3.6 in 1993 to 5.8 in 1998; P < .001), in line with guideline recommendations. Rates of switching to antidepressants increased over time (8.5% in 1993 to 10.2% in 1998; P < .001), whereas switching to barbiturates was consistently low (0.12%; P = .403). CONCLUSION: The prevalence of benzodiazepine therapy for older people in Ontario has steadily declined between 1993 and 1998. There is a trend of dispensing relatively more short-acting than long-acting benzodiazepines and of replacing benzodiazepines with antidepressants in older people without a remarkable increase in barbiturate consumption. These findings suggest that, without undue regulation, physicians are making progress in the prescribing of benzodiazepine therapy on the basis of current knowledge available. PMID- 11890494 TI - Racial and state differences in the designation of advance directives in nursing home residents. AB - OBJECTIVES: To examine racial and state differences in the use of advance directives and surrogate decision-making in a nursing home population. DESIGN: A retrospective cohort study. SETTING: Nursing homes in the states of California (CA), Massachusetts (MA), New York (NY), and Ohio (OH). PARTICIPANTS: Nursing home residents: 130,308 in CA, 59,691 in MA, 112,080 in NY, and 98,954 in OH. MEASUREMENTS: Minimum Data Set information concerning resident race and whether or not residents have a living will (LW), a do not resuscitate (DNR) order, or a surrogate decision-maker (SDM). RESULTS: The proportion of LWs, DNR orders, and SDMs varied significantly (P < .0001) by racial categories in each state. In general, whites were distinctly different from other racial categories. Whites were significantly more likely to have a LW (odds ratio (OR) = 1.9 (CA), OR = 2.2 (NY), OR = 4.9 (OH)), a DNR order (OR = 2.4 (CA), OR = 2.4 (MA), OR = 3.3 (NY), OR = 3.2 (OH)), and a SDM (OR = 1.1 (CA), OR = 1.2 (NY), OR = 1.6 (OH)) than were nonwhites, after adjusting for potentially confounding factors. Significant state differences (P < .0001) were observed in LWs, DNR orders, and SDMs and were most pronounced in residents of Ohio, who were significantly more likely to have a LW than were residents in other states (OR = 9.3). CONCLUSIONS: Various resident characteristics explain some of the racial differences, although whites are still more likely to have a LW, a DNR order, or an SDM independent of various resident characteristics included in the adjusted analyses. This pattern is observed in all states, although the ORs varied by state. Some of these differences may be due to distinct cultural approaches to end-of-life care and lack of knowledge and understanding of advance directives. The distinctly higher rates of LWs among all racial groups in Ohio than in other states suggest that states can potentially increase the use of advance directives through intervention. PMID- 11890495 TI - Physician-patient congruence regarding medication regimens. AB - OBJECTIVES: To characterize the degree of disparity between physicians' perceptions of older patients' medication regimen and patients' perceptions of their regimen. DESIGN: Prospective observational. Physicians and patients were blinded at index visit; after, trained medical students made home visits, collecting information about medications that was compared with physician questionnaires. SETTING: Community family medicine residency program. PARTICIPANTS: Patients age 65 and older presenting for routine visit, taking at least four prescription medications, and seen by index physician three or more times in the past year. Physicians were family medicine faculty and second- and third-year residents. MEASUREMENTS: Fifty physician-patient pairs were analyzed. Average age was 75 (standard deviation (SD) +/-5.5); patients averaged 7.0 prescription medications (range 3-17, SD +/-2.89). Three hundred seventy-five prescription medications were identified; the most commonly prescribed were antihypertensives (134/375; 36%). RESULTS: Congruence, defined as agreement between physician and patient regarding all prescription medications, dosages, and frequency, averaged 70% for faculty (range 53-89%) and 58% for residents (range 41-81%) (P = .08). Fourteen percent (7/50) demonstrated complete congruence; 74% (37/50) had at least one medication that either the physician was unaware the patient was taking or the physician thought the patient was taking but that was not part of the patient's regimen; 12% (6/50) had dose and/or frequency discrepancies. CONCLUSIONS: This study demonstrates significant disparity in a population where it is crucial for healthcare providers and patients to be in close agreement about intended medication regimens. PMID- 11890496 TI - Primary myelodysplasia: management and outcome at 3 years in 45 patients age 65 and older. AB - OBJECTIVES: To study myelodysplasia in patients age 65 and older. SETTING: A French university hospital. PARTICIPANTS: Forty-five patients age 65 and older with a diagnosis of myelodysplasia made in the hospital between January 1993 and December 1998. MEASUREMENTS: Clinical presentation, initial hematological features, type of myelodysplasia (French American British classification), treatment, and evolution at 36 months were studied. RESULTS: The mean age of the group was 78. Anemia was the initial hematological feature in 30 patients out of 45 and was symptomatic in 24 patients. Refractory anemia was diagnosed in 20 patients; 11 patients presented with refractory anemia with excess blast cells, eight with chronic myelomonocytic leukemia. Thirty-one patients received erythrocyte transfusions, no patient received chemotherapy. At 36 months, 68.8% of the patients were dead. The median survival (22 months) was lower than in other reported series even in types of myelodysplasia with a classically better prognosis. CONCLUSION: Myelodysplasia is probably underdiagnosed in older people and has a poor prognosis (median survival 22 months), and no effective treatment is available in older patients. PMID- 11890497 TI - Medical mistakes and older patients: admitting errors and improving care. AB - The topic of medical errors has received substantial professional and public attention over the past few years. However, very little of this attention has focused specifically on the implications of this problem for older patients or on the healthcare professionals and settings specifically serving them. This article examines some of the most salient of those implications, with particular emphasis on the physician's ethical duty to admit and address errors. Practical obstacles, including physicians' legal anxieties, to admitting and thereby reducing and mitigating medical errors are outlined, along with potential strategies for effectively addressing and overcoming those barriers. PMID- 11890498 TI - Health care for older persons: a country profile--Lebanon. AB - Lebanon is a small country, comparable in size to New Hampshire. It is currently estimated that 8% of the population is age 65 and older. The Lebanese population has witnessed a clear demographic transition in the past few decades. Our culture demands respect for older people and values highly the natural bonds of affection between all members of the family. The healthcare system in the country is an adaptation of the European model. Despite the large number of physicians (approximately 10,000) there is a shortage of primary care and geriatric physicians. There are 36 nursing homes in Lebanon, with a total of 6,000 beds, but most of them are understaffed, with the exception of three nursing homes that offer relatively comprehensive services including rehabilitative, preventive, and curative services. The Ain Wizen Elderly Care Centre is well recognized for the program it operates for older people, which is a good model for the region and for Lebanon in terms of services, training, and research. Demographic changes and social and economic developments in Lebanon have created new realities in the unprecedented growth of the older population. Lebanon, like other developing countries, needs to define the policies and programs that will reduce the burden of an aging population on its society and economy. There is a need to ensure the availability of health and social services for older persons and to promote older persons' continuing participation in a socially and economically productive life. PMID- 11890499 TI - Ethnicity and cognitive performance among older African Americans, Japanese Americans, and Caucasians: the role of education. AB - This cross-sectional analysis evaluated the association between ethnicity and cognitive performance and determined whether education modifies this association for nondemented older people (103 African Americans, 1,388 Japanese Americans, 2,306 Caucasians) in a study of dementia incidence. African Americans scored lower (median 89 out of 100) than Japanese Americans (93) and Caucasians (94) on the Cognitive Abilities Screening Instrument (CASI). Education affected CA PMID- 11890500 TI - Establishing a case-finding and referral system for at-risk older individuals in the emergency department setting: the SIGNET model. AB - Older emergency department (ED) patients have complex medical, social, and physical problems. We established a program at four ED sites to improve case finding of at-risk older adults and provide comprehensive assessment in the ED setting with formal linkage to community agencies. The objectives of the program are to (1) improve case finding of at-risk older ED patients, (2) improve care planning and referral for those returning home, and (3) create a coordinated network of existing medical and community services. The four sites are a 1,000 bed teaching center, a 700-bed county teaching hospital, a 400-bed community hospital, and a health maintenance organization (HMO) ED site. Ten community agencies also participated in the study: four agencies associated with the hospital/HMO sites, two nonprofit private agencies, and four public agencies. Case finding is done using a simple screening assessment completed by the primary or triage nurse. A geriatric clinical nurse specialist (GCNS) further assesses those considered at risk. Patients with unmet medical, social, or health needs are referred to their primary physicians or to outpatient geriatric evaluation and management centers and to community agencies. After 18 months, the program has been successfully implemented at all four sites. Primary nurses screened over 70% (n = 28,437) of all older ED patients, GCNSs conducted 3,757 comprehensive assessments, participating agency referrals increased sixfold, and few patients refused the GCNS assessment or subsequent referral services. Thus, case finding and community linkage programs for at-risk older adults are feasible in the ED setting. PMID- 11890501 TI - Anabolic agents to treat osteoporosis in older people: is there still place for fluoride? Fluoride for treating postmenopausal osteoporosis. PMID- 11890502 TI - Older persons in the emergency medical care system. PMID- 11890503 TI - Ethnicity and dementia. PMID- 11890504 TI - Night functional dependency index. PMID- 11890505 TI - Recombinant erythropoietin treatment of anemia in older adults. PMID- 11890506 TI - Evaluation of pain in cognitively impaired individuals. PMID- 11890507 TI - Hearing disability in older adults: patient and doctor delay in primary medical care. PMID- 11890508 TI - State practice variation in the use of tube feeding for nursing home residents with severe cognitive impairment. PMID- 11890509 TI - Hospitalization outcomes. PMID- 11890510 TI - Comparison of in-patient and out-patient penile prosthesis surgery. AB - Between December 1996 and December 1998, 79 inflatable penile implant insertions have been performed at our institution by a single surgeon. The objective of this analysis was to compare our in-patient and out-patient experience with penile prosthesis insertion with respect to ease of performance and complication profiles. Data was collected in a prospective manner for both groups (in-patient, n = 33 and out-patient, n = 46). The two groups were compared with respect to intra-operative blood loss, operative time, time lost from work, narcotic use and complication rates. Both groups of patients experienced similar operative blood loss, essentially identical operative times, time lost from work and narcotic use. Most importantly, overall complication rates were 6% for the in-patient group and 4% for the out-patient group. Inflatable penile implant surgery is feasible in an ambulatory surgical setting. There is no difference in complication rates, loss of time from work, or intra-operative and post-operative course. Furthermore, there is a significant saving at our institution by performing the procedure in an out-patient fashion. In-patient prosthetic surgery is reserved for secondary procedures following a prior implant infection or primary implants in men with significant co-morbidities that require in-patient postoperative monitoring. PMID- 11890511 TI - Erectile dysfunction occurs following substantia nigra lesions in the rat. AB - Erectile function was assessed 6 weeks following uni- and bilateral injections of 6-hydroxydopamine in the substantia nigra nucleus of the brain. Behavioral apomorphine-induced penile erections were reduced (5/8) and increased (3/8) in uni- and bilateral lesioned animals. Intracavernous pressures, following electrical stimulation of the cavernous nerve, decreased in lesioned animals. Lesions of the substantia nigra were confirmed by histology. Concentration of dopamine and its metabolites were decreased in the striatum of substantia nigra lesioned rats. Lesions of the substantia nigra are therefore associated with erectile dysfunction in rats and may serve as a model to study erectile dysfunction in Parkinson's disease. PMID- 11890512 TI - Assessment of the efficacy and safety of Viagra (sildenafil citrate) in men with erectile dysfunction during long-term treatment. AB - Long-term efficacy and safety of sildenafil was assessed in 1008 patients with erectile dysfunction (ED) enrolled in four flexible-dose (25 - 100 mg), open label, 36- or 52-week extension studies. After 36 and 52 weeks, 92% and 89% of patients felt that treatment with sildenafil had improved their erections. Responses to a Sexual Function Questionnaire indicated that 52 weeks of sildenafil treatment resulted in clinically significant improvements in the duration and firmness of erections, overall satisfaction with sex life, and the frequency of stimulated erections. Commonly reported adverse events (AEs) were headache, flushing, dyspepsia, and rhinitis, which were generally mild to moderate. Reports of abnormal vision were consistent with previous clinical trials. The occurrence of treatment-related cardiovascular AEs, such as hypertension, tachycardia, and palpitation, was <1%. Discontinuations due to treatment-related AEs were low (2%). Long-term therapy does not diminish the efficacy of sildenafil in patients with ED and remains well tolerated. PMID- 11890513 TI - Creative-dynamic image synthesis: a useful addition to the treatment options for impotence. AB - In contrast to the impressive advances made in somatic research on erectile dysfunction, psychogenic erectile dysfunction is usually treated as a monolithic block. In this study, we evaluated the erectogenic power of creative-dynamic image synthesis in men with psychogenic erectile dysfunction. Sixty-nine men with a mean age of 46 y, suffering from erectile dysfunction of no known organic cause, were entered in a placebo-controlled study in which the erectogenic power of imagination, yohimbine and a placebo were compared. There was a significant difference between the subjective results of creative-dynamic image synthesis (75% increase of potency) and those achieved through treatment with the drug yohimbine (55% increase in potency) and with a placebo (30% increase in potency). Creative-dynamic image synthesis is a potent initiator of erections in men with psychogenic erectile dysfunction, has no known side effects and is very cost effective. PMID- 11890514 TI - Erectile dysfunction is a marker for cardiovascular complications and psychological functioning in men with hypertension. AB - The aim of this study was to investigate the incidence of cardiovascular complications in hypertensive patients with erectile dysfunction (ED). An anonymous questionnaire was mailed to 467 and received from 104 hypertensive male patients. Despite the low response rate of 22%, the following interesting findings could be observed: 70.6% of the patients who responded suffered from ED. The hypertensive patients with ED had significantly higher prevalence of cardiovascular complications (P < 0.05). The correlation between depression and low quality of life as well as between ED and low sexual satisfaction was also statistically significant (P = 0.05). ED in hypertensive patients can be considered as a marker for cardiovascular complications in this patient group. PMID- 11890515 TI - The phosphodiesterase inhibitory selectivity and the in vitro and in vivo potency of the new PDE5 inhibitor vardenafil. AB - We investigated the potency and the selectivity profile of vardenafil on phosphodiesterase (PDEs) enzymes, its ability to modify cGMP metabolism and cause relaxation of penile smooth muscle and its effect on erections in vivo under conditions of exogenous nitric oxide (NO) stimulation. PDE isozymes were extracted and purified from human platelets (PDE5) or bovine sources (PDEs 1, 2, 3, 4 and 6). The inhibition of these PDEs and of human recombinant PDEs by vardenafil was determined. The ability to potentiate NO-mediated relaxation and influence cGMP levels in human corpus cavernosum strips was measured in vitro, and erection-inducing activity was demonstrated in conscious rabbits after oral administration together with intravenous doses of sodium nitroprusside (SNP). The effects of vardenafil were compared with those of the well-recognized PDE5 inhibitor, sildenafil (values for sildenafil in brackets). Vardenafil specifically inhibited the hydrolysis of cGMP by PDE5 with an IC50 of 0.7 nM (6.6 nM). In contrast, the IC50 of vardenafil for PDE1 was 180 nM; for PDE6, 11 nM; for PDE2, PDE3 and PDE4, more than 1000 nM. Relative to PDE5, the ratios of the IC50 for PDE1 were 257 (60), for PDE6 16 (7.4). Vardenafil significantly enhanced the SNP-induced relaxation of human trabecular smooth muscle at 3 nM (10 nM). Vardenafil also significantly potentiated both ACh-induced and transmural electrical stimulation-induced relaxation of trabecular smooth muscle. The minimum concentration of vardenafil that significantly potentiated SNP-induced cGMP accumulation was 3 nM (30 nM). In vivo studies in rabbits showed that orally administered vardenafil dose-dependently potentiated erectile responses to intravenously administered SNP. The minimal effective dose that significantly potentiated erection was 0.1 mg/kg (1 mg/kg). The selectivity for PDE5, the potentiation of NO-induced relaxation and cGMP accumulation in human trabecular smooth muscle and the ability to enhance NO-induced erection in vivo indicate that vardenafil has the appropriate properties to be a potential compound for the treatment of erectile dysfunction. Vardenafil was more potent and selective than sildenafil on its inhibitory activity on PDE5. PMID- 11890516 TI - Prevalence of Peyronie's disease in men over 50-y-old from Southern Brazil. AB - The pathogenesis of Peyronie's disease still remains an enigma and few epidemiological studies are available. The purpose of this study was to determine the prevalence of Peyronie's disease in males older than 50 y. From 26 to 30 July 1998, 1071 men attended the 'Prostate Cancer Awareness Week of Santa Casa Hospital, Porto Alegre, Brazil'. In the prostate exam they also consented to be screened for Peyronie's disease. They underwent the 5-item International Index of Erectile Function (IIEF-5) questionnaire for evaluation of the erectile condition. The presence of a well-defined plaque in the penis was the diagnostic criterion for Peyronie's disease. The men were examined by five senior residents, under supervision by the staff Urologist. Men younger than 50 y as well as patients under intracavernous injection therapy for erectile dysfunction were excluded from the study. Chi2 test was used for statistical analysis. Nine hundred and fifty-four (89.1%) out of the 1071 men with a mean age of 62 y (ranging from 52 to 77) were included in the study. Peyronie's disease plaques were found in 35 men (3.67%). Eight hundred and forty-five (88.6%) were Caucasians. There was no significant statistical difference regarding age (P > 0.05). The presence of erectile dysfunction in the men with Peyronie's disease and without this condition, was 68.6% and 53.5%, respectively (P > 0.05). From this data we can conclude that the prevalence of Peyronie's disease is higher than in formerly reported studies. Further observations should be carried out in different communities and in other groups of patients in order to confirm our results. PMID- 11890517 TI - Spring balance evaluation of the ischiocavernosus muscle. AB - We studied the voluntary contractile activity of the ischiocavernosus muscle (ICM) in 21 sexually potent and 97 erectile dysfunction (ED) subjects using a spring balance. A strap was placed around the coronal grove of the glans penis and tensioned with the spring balance. Subjects were asked and encouraged to contract the ICM against the spring balance. We evaluated the length of stroke, duration of contraction, and maximum contractile force. The length of stroke, duration of contraction, and maximum contractile force showed statistically significant differences between potent and ED subjects. Diagnosed psychogenic ED and arteriogenic ED showed higher contractile activity than cavernous ED and neurogenic ED. Our results corresponded to those of previous studies that have urged consideration of the role of the ICM during the process of erection in animal experiments and in human electrophysiological studies. The spring balance evaluation is a useful, inexpensive method for evaluating the ICM. PMID- 11890518 TI - Does bicycling contribute to the risk of erectile dysfunction? Results from the Massachusetts Male Aging Study (MMAS). AB - An association between bicycling and erectile dysfunction (ED) has been described previously, but there are limited data examining this association in a random population of men. Such data would incorporate bicyclists with varied types of riding and other factors. Data from the Massachusetts Male Aging Study (MMAS) were utilized to examine the association between bicycling and ED. Logistic regression was used to test for an association, controlling for age, energy expenditure, smoking, depression and chronic illness. Bicycling less than 3 h per week was not associated with ED and may be somewhat protective. Bicycling 3 h or more per week may be associated with ED. Data revealed that there may be a reduced probability of ED in those who ride less than 3 h per week and ED may be more likely in bikers who ride more than 3 h per week. More population-based research is needed to better define this relationship. PMID- 11890520 TI - Bisphenol A inhibits penile erection via alteration of histology in the rabbit. AB - Despite extensive research into the toxicity of bisphenol A (BPA), no report of its effect on erectile function exists. We performed this study to investigate the effect of BPA on erectile function. New Zealand white rabbits were treated intraperitoneally with 150 mg/kg of BPA every other day for 12 days (cumulative dose of 900 mg/kg). Four and 8 weeks after administration of BPA, the contractions and relaxation of cavernosal tissue strips were significantly suppressed in the BPA-treated animals compared to the control animals. Histologically, thickening of tunica albuginea, subtunical deposition of fat and decreased sinusoidal space with consequent increase of trabecular smooth muscle content were observed in the BPA-treated animals. These results suggest that xenoestrogen BPA may affect the erectile function through evident histological changes of the penis. PMID- 11890519 TI - Effects of oral phentolamine, taken before sleep, on nocturnal erectile activity: a double-blind, placebo-controlled, crossover study. AB - The objective of this study was to determine the effects of oral phentolamine, administered before sleep, on nocturnal penile erectile activity of men with mild to moderate erectile dysfunction (ED). We studied five patients with mild to moderate ED (mean age 34.8 +/- 8.13 and mean duration of ED 31.8 +/- 23.5 months), in a double-blind, placebo-controlled, crossover study. All patients received oral phentolamine (Vasomax) at a dose of 40 mg and placebo for three consecutive nights respectively and were submitted to nocturnal penile tumescence and rigidity monitoring (NPTR) with the Rigiscan device. NPTR parameters of the two 3-night recordings were evaluated and compared. Administration of oral phentolamine before sleep was associated with a statistically significant increase in the number of erectile events with rigidity > or = 60% lasting > or = 10 min (P = 0.02), as well as the rigidity activity units (RAU) value per hour sleep, both at the base (P = 0.023) and the tip of the penis (P = 0.019). The number of events as measured by Rigiscan software (20% change in circumference), as well as tumescence activity units (TAU)/h values did not show any statistical difference. No adverse effects were recorded. It is concluded that oral phentolamine administered before sleep enhanced NPTR parameters associated with the quality of the erectile events. Such results provide a pathway for the development of a prevention strategy for ED. Future studies will elucidate whether vasoactive agents taken on a regular basis before sleep, can prevent ED in men at risk, protecting also minimally and moderately impotent patients to become moderately and severely impotent respectively. PMID- 11890521 TI - Respiratory syncytial virus infection induces matrix metalloproteinase-9 expression in epithelial cells. AB - Increased gelatinolytic activity was observed in respiratory syncytial virus (RSV)-infected HEp-2 cells by using zymography. The anti-matrix metalloproteinase 9 (MMP-9) antibody specifically reduced the gelatinolytic activity suggesting that the increased gelatinolytic activity was due to the MMP-9. It was also supported by the results from immunofluorescent staining, treatment of MMP inhibitors, and RSV infection of the cell clones that were transfected with plasmids to express more MMP-9 and tissue type inhibitor of metalloproteinase-1 (TIMP-1). The gelatinolytic activity of extracellular MMP-9 in RSV-infected HEp-2 cells increased 1.5 +/- 0.2 fold compared with the control (p < 0.01). Cell surface MMP-9 expression was also clearly detected by immunofluorescent staining. Treatment with 1,10-phenanthroline (0.05 mM), ethylenediamine-tetraacetate (EDTA) (1.5 mM), and penta-O-galloyl-beta-D-glucose (PGG) (3.3 microM) inhibited RSV multiplication as well as syncytia formation. Furthermore, the average syncytia size increased when the cells expressing more MMP-9 were infected by RSV. In contrast, syncytia formation was inhibited in the cells manipulated to express TIMP-1. Thus, this study concludes that although RSV infection induces MMP-9, which can enhance the syncytia formation leading to RSV multiplication and spread it can be inhibited by MMP inhibitors. PMID- 11890522 TI - Molecular characterisation of two mumps virus genotypes circulating during an epidemic in Lithuania from 1998 to 2000. AB - An epidemic of mumps in Lithuania started in December 1998 and continued until May 2000. The total registered number of cases was about 11.000 of a total of 3,7 million inhabitants in Lithuania (29,7 cases/10,000). Virus-containing samples were collected from 80 patients treated at the hospital of Kaunas from October 1999 until the end of the epidemic. Out of the 80 patients with parotitis, meningitis was observed in 11 patients and orchitis in 22 of 69 male patients. Twenty-seven virus strains were genotyped by nucleotide sequencing of the small hydrophobic (SH) protein gene, and the 57 amino acid sequences of the gene were deduced. Twenty-five virus strains belonged to the C genotype and two were of the D genotype. By phylogenetic analysis the virus strains causing meningitis grouped in a separate cluster, designated C1, within the C genotype. Another group of ten of the 25 genotype C strains exhibited an amino acid triplet at amino acid positions 28 to 30 of the protein, consisting of valine, alanine and serine, instead of the previously recognised valine, valine and serine combination of genotype C. The amino acid alanine at position 29 was found in combination with the amino acid serine at position 48. This variant was designated C2 and it was associated with parotitis. The amino acid alanine at position 29 and serine in position 48 of the C2 genotype may constitute a marker of low neurovirulence compared to other genotype C strains. PMID- 11890523 TI - In vitro anti-influenza virus activity of synthetic humate analogues derived from protocatechuic acid. AB - Two humic-like substances, the oxidative polymer of protocatechuic acid (OP-PCA) and humic acid inhibit the in vitro replication of influenza virus A/WSN/33 (H1N1) in Madin-Darby canine kidney (MDCK) cells at concentrations of no cytotoxicity. The 50% inhibitory concentration (IC50) for OP-PCA was 6.59 +/- 1.02 microg/ml when the compound was added at the stage of viral adsorption. When OP-PCA was added after virus adsorption, the IC50 was 53.27 +/- 8.12 microg/ml. The IC50 for humic acid was 48.61 +/- 7.32 microg/ml and 55.27 +/- 5.46 microg/ml respectively when the compound was added at the stage of viral adsorption or post adsorption. In spite of structural resemblance of these two compounds, they exhibit different actions of anti-flu. The OP-PCA inhibits virus-induced hemagglutination and low pH-induced cell-cell fusion. Humic acid inhibits the endonuclease activity of viral RNA polymerase. The monomer of PCA shows no inhibition on influenza virus replication. PMID- 11890524 TI - Role of intrabursal T cells in infectious bursal disease virus (IBDV) infection: T cells promote viral clearance but delay follicular recovery. AB - Infectious bursal disease virus (IBDV) induces an acute, highly contagious immunosuppressive disease in young chickens. We examined the role of T cells in IBDV-induced immunopathogenesis and tissue recovery. T cell-intact chickens and birds compromised in their T cell function by a combination of surgical thymectomy and Cyclosporin A treatment (Tx-CsA) were infected with an intermediate vaccine strain of IBDV (Bursine 2, Fort Dodge). Our data revealed that functional T cells were needed to control the IBDV-antigen load in the acute phase of infection at 5 days post infection. The target organ of IBDV, the bursa of Fabricius, of Tx-CsA-birds had a significantly higher antigen load than the one of T cell-intact birds (P < 0.05). Tx-CsA-treatment abrogated the IBDV induced inflammatory response and significantly (P < 0.05) reduced the incidence of apoptotic bursa cells and the expression of cytokines such as interleukin 2 (IL-2) and interferon-gamma (IFN-gamma) in comparison to T cell-intact birds. T cell-released IL-2 and IFN-gamma may have mediated the induction of inflammation and cell death in T cell-intact birds. The IBDV-induced upregulation of tumor necrosis like-factor (TNF) expression was comparable between T cell-intact and Tx CsA-birds. Tx-CsA-birds showed a significantly faster resolution of IBDV-induced bursa lesions than T cell-intact birds (P < 0.05). This study suggests that T cells modulate IBDV pathogenesis in two ways: a) they limit viral replication in the bursa in the early phase of the disease at 5 days post infection, and b) intrabursal T cells promote bursal tissue damage and delay tissue recovery possibly through the release of cytokines and cytotoxic effects. PMID- 11890525 TI - Productive infection of a mink cell line with porcine endogenous retroviruses (PERVs) but lack of transmission to minks in vivo. AB - Porcine endogenous retroviruses (PERVs) are considered a special risk for xenotransplantation because they are an integral part of the porcine genome and are able to infect cells of numerous species including humans in vitro. Among these cells, the mink lung epithelial cell line Mv1Lu could be productively infected with PERV. Provirus integration was detected by PCR, expression of viral proteins was shown by immunostaining and reverse transcriptase was detected in cell supernatants. PERV produced from mink cells could infect both, uninfected mink Mv1Lu cells and uninfected human 293 cells, with considerably higher virus production by human cells. Typical type C retroviruses were observed in PERV infected mink cells using electron microscopy together with numerous multivesicular body (MVB)-like structures containing small virus-like particles, not present in uninfected mink cells. These MVBs could be stained with PERV specific serum. In an attempt to establish a small animal model, PERV grown on mink cells was inoculated into adult and newborn American minks. Neither antibody production against PERV nor integration of viral DNA or production of viral proteins in tissues of different organs could be detected 12 weeks post virus inoculation, indicating that PERV infection had not occurred. PMID- 11890526 TI - Generation of a life-expanded rhesus monkey fibroblast cell line for the growth of rhesus rhadinovirus (RRV). AB - RRV, the rhesus macaque equivalent to HHV-8 or kaposi's sarcoma-associated herpesvirus (KSHV) was recently isolated from a simian immunodeficiency virus (SIV) infected macaque with a lymphoproliferative disorder. The growth of RRV in tissue culture requires propagation of primary rhesus monkey fibroblasts (RFs). In an effort to extend the life of these primary cells in tissue culture, the catalytic subunit of telomerase (hTERT) was introduced into RF cells using a recombinant retrovirus. This new cell line, Telo-RFs, have currently been passed in tissue culture over 80 times compared to a maximum passage number of 38 for wild type RFs, remain fully permissive for RRV DNA replication and production of infectious virus. Viral gene expression of immediate-early and early RNA transcripts was virtually identical to that observed in wild-type (wt) RFs. In addition, transfection experiments show that telo-RFs are easily and more efficiently transfected than wtRFs. PMID- 11890527 TI - Expression of the simian varicella virus glycoprotein L and H. AB - Simian varicella is used as a model to investigate varicella-zoster virus pathogenesis and to evaluate antiviral therapies. In this study, the simian varicella virus (SVV) glycoprotein L (gL) was characterized along with its association with glycoprotein H (gH). The SVV gL gene encodes a predicted 175 amino acid polypeptide that shares 43.5% and 27.9% amino acid identity with the VZV gL and HSV-1 gL, respectively. The SVV gL polypeptide sequence lacks a consensus glycosylation site and a typical signal sequence, but does possess an endoplasmic reticulum targeting sequence found commonly in chaperone proteins. Transcriptional analysis indicated that the SVV gL and the uracil DNA glycosylase (UDG) genes share a common 5' RNA start site and are co-expressed on a 2.0 kb transcript. gL and gH expression in SVV-infected Vero cells was demonstrated by immunofluorescence and immunoprecipitation analyses using specific antisera generated against gL and gH peptides. Similar to other herpesvirus gH and gL homologs, the SVV gL and gH form a complex within infected cells. gL and gH transcripts and antigens were detected in tissues of monkeys with acute simian varicella. The simian varicella model offers an opportunity to investigate the role of the gL and gH in viral pathogenesis. PMID- 11890528 TI - Genomic variability in Potato potyvirus Y (PVY): evidence that PVY(N)W and PVY(NTN) variants are single to multiple recombinants between PVY(O) and PVY(N) isolates. AB - Fourteen Potato virus Y (PVY) isolates representative of PVY(O), PVY(N), PVY(NTN) and PVY(N)W groups were characterised at genomic level. Restriction fragment length polymorphism study (RFLP) of each gene of these isolates and sequencing of the first 2700 nucleotides of two PVY(N)W isolates were performed. A mosaic structure was revealed in PVY(N)W and PVY(NTN) genomes, which showed either PVY(O) or PVY(N)-like sequences, depending on the particular gene. Indeed, starting from the 5'-end, these isolates showed a switching, from PVY(N)- to PVY(O)-like sequence, in the HC-Pro C-terminal region. Reversion to PVY(N)-like sequence was also revealed in the NIa N-terminal area of PVY(NTN) isolates, followed by a switching back to a PVY(O)-like sequence in the CP gene. Lastly, some PVY(N)W isolates showed a switching from PVY(O)- to PVY(N)-like sequence in the P1 N-terminal part, thus separating our PVY(N)W isolates into two subgroups. All these apparent recombination events were shown by statistical analysis. Comparison of molecular traits with pathogenic properties of our isolates suggested that the HC-Pro protein is involved in induction of necrosis in tobacco leaves, and the NIa, NIb and/or CP protein in necrosis in potato tubers. Nevertheless, multiple recombination events observed in the PVY(NTN) genome may play a role in the latter phenomenon. PMID- 11890529 TI - Molecular and serological relationships of Spartina mottle virus (SpMV) strains from Spartina spec. and from Cynodon dactylon to other members of the Potyviridae. AB - Spartina mottle virus (SpMV) was first reported 1980 and classified by physical and biological properties as a tentative member of the genus Rymovirus in the Potyviridae. This genus was recently separated into two genera: Rymovirus and Tritimovirus. Now the sequence of the 3'-terminal part of the genome of SpMV was determined. Additionally a virus isolate originating from Cynodon dactylon in Italy was cloned and sequenced. This Assisi-isolate shared 87.5% amino acid sequence identity with SpMV. The high degree of identity and their close serological relationship indicate that SpMV and Assisi-isolate have to be regarded as different strains of one virus. The Assisi-isolate should be designated as SpMV-AV. Comparing the C-terminal part of the ORF of several Potyviridae the sequences of SpMV strains were more similar to those of the genera Rymovirus and Potyvirus than to the genera Tritimovirus, Macluravirus, Bymovirus and Ipomovirus. The comparisons revealed identities of less than 32% for the CP and 37% for the 3'-NIb/CP region, indicating that SpMV can not be classified to any of the established genera. The results of serological tests support a separate position of SpMV in the Potyviridae. We propose to introduce the name Sparmovirus for the new genus. PMID- 11890530 TI - Isolation and phylogeny of endogenous retrovirus HERV-F family in Old World monkeys. Brief report. AB - A new human endogenous retroviral family (HERV-F) has recently been identified from human chromosome 7q31.1-q31.3 that was identical to the XA34 cDNA clone isolated from a human glioma cDNA library with an ERV-9 env probe. We investigated pol fragments of the HERV-F family from the Old World monkeys (crab eating monkey, African green monkey, and baboon) and analyzed these with the HERV F (Hu-XA34). Fifteen pol fragments of the HERV-F family were detected from the Old World monkeys. They showed a high degree of sequence similarity (81-99%) with that of the HERV-F (Hu-XA34). Phylogenetic analysis of pol fragments with those of the human genome distinctively showed five groups, indicating that HERV-F family could be amplified at least five times after the original integration into the monkey genome or represent integration events independently during primate evolution. PMID- 11890531 TI - Serological and genetic characterization of newly isolated Peaton virus in Japan. Brief report. AB - The viruses were isolated from the blood of sentinel cattle and Culicoides biting midges in the Kyushu district, southwestern Japan, in 1999 and identified by neutralization tests as Peaton (PEA) viruses. Before this study, PEA virus had been isolated in Australia only. The nucleotide identity of the nucleocapsid (N) protein encoded by the S segment ranged from 91.1 to 91.6% between the Australian and Japanese strains. A phylogenetic analysis of the N protein sequence revealed that the PEA virus strains are closely related to Aino (AIN) virus and suggested reassortment events for PEA and AIN viruses. PMID- 11890532 TI - Molecular characterisation and evolutionary relationships of a potyvirus infecting Crotalaria in Brazil. Brief report. AB - The 3' terminal genomic region of a potyvirus causing mosaic disease in several Crotalaria species has been cloned and sequenced. Comparisons of the nucleotide and deduced amino acid (aa) sequences of the cloned cDNA with those from other potyviruses show that the Crotalaria-infecting virus (designated Crotalaria mosaic virus; CrMV) is closely related to Cowpea aphid-borne mosaic virus (CABMV). Maximum identity (95.4%) at the coat protein (CP) aa level was observed between CrMV and a Brazilian strain of CABMV. Phylogenetic analyses derived from the sequence alignments of the CP and 3' untranslated region confirmed the identification of CrMV as a strain of CABMV and the name CABMV-Cr is suggested. PMID- 11890533 TI - Characterisation of some carla- and potyviruses from bulb crops in China. Brief report. AB - Conserved carla- and potyvirus primers were used in RT-PCR to amplify virus fragments from garlic and other bulb crops in China and the fragments were subsequently sequenced and compared in phylogenetic analyses. Garlic plants from Henan, Hubei, Jiangsu, Shangdong and Yunnan provinces all contained at least one isolate each of Garlic latent virus (genus Carlavirus), Onion yellow dwarf virus (OYDV, genus Potyvirus) and Leek yellow stripe virus (LYSV, genus Potyvirus). The complete sequence of a Zhejiang isolate of LYSV was also determined, providing the first complete sequence of this virus. The genome was 10142 nucleotides long excluding the poly(A) tail and had the typical features of the genus Potyvirus, although some of the amino acids surrounding the polyprotein cleavage sites were unusual. Shallot yellow stripe virus (SYSV) was amplified from bunching onion (Allium fistulosum var. caespitosum) in Zhejiang province, providing the first record of SYSV in China. Lily mottle virus was amplified from dragon-teeth lily (Lilium brownii var. viridulum). PMID- 11890534 TI - Detection of antibodies to Borna disease virus (BDV) in Turkish horse sera using recombinant p40. Brief report. AB - The nucleoprotein of Borna disease virus (BDV-p40) was produced in a Baculovirus expression system using sf9 cells. The purity and specificity of the recombinant p40 was confirmed by SDS-PAGE and immunoblotting. The recombinant p40 was used in an ELISA to screen horse sera in Turkey. For this, 323 horses from selected cities in the Marmara region of Turkey were examined clinically and serum was collected from each. All horses were clinically healthy except for a few with wounds on the skin. Antibodies to BDV were detected in the sera of 82 (25%) of 323 horse sera. Six sera were selected that had low, medium or high OD values by ELISA and were analysed by Western blotting. All reacted specifically with p40 at a dilution of 1 in 1000. This is the first report of the detection of Borna disease in Turkey and needs further molecular biological investigations to compare the Turkish strains with those strains detected in Europe. PMID- 11890535 TI - Nucleotide sequence of a Japanese isolate of Squash mosaic virus. Brief report. AB - The complete nucleotide sequence of the RNA-1 of Squash mosaic virus (SqMV) was determined using a Japanese isolate (Y-SqMV). The sequence consisted of 5865 nucleotides excluding the poly (A) at the 3' terminus and contained a single long open reading frame with a coding capacity for a protein of Mr209971. Analysis of the deduced amino acid sequence suggested a genomic organization typical of comoviruses. The nucleotide sequence of the RNA-2 of Y-SqMV was also determined and compared with the SqMV isolates from the United States. The larger and smaller capsid protein (CP) coding region was compared to those of K-SqMV and Z SqMV, which represent two subgroups of SqMV. The larger CP gene of Y-SqMV showed 93.0% and 88.0% identities with those of K-SqMV and Z-SqMV, respectively at the nucleotide level. The smaller CP gene of Y-SqMV was 94.1% and 88.4% identical with those of K-SqMV and Z-SqMV. The results suggested that the Japanese SqMV isolate (Y-SqMV) is distinct from those in the United States, and might represent a third subgroup. PMID- 11890536 TI - Proceedings of the 10th European Workshop Conference on Bacterial Protein Toxins (ETOX). June 24-29, 2001, Bohon, Belgium. PMID- 11890537 TI - Molecular and cellular basis of the infection by Listeria monocytogenes: an overview. AB - This review rather than covering the whole field, intends to highlight the particularly interesting properties of some proteins involved in the infection by Listeria monocytogenes and the general interest in some of the approaches used to analyze the molecular and cellular basis of the infection. After an introduction to the bacterium and to the disease, a description of the infection at the cellular level will be given. The specific features of the pore-forming toxin listeriolysin O, as a protein particularly well adapted to the intracellular lifestyle of L. monocytogenes will be discussed. By describing in detail how the bacterium moves inside cells, particular attention will be given to show how addressing this issue has provided key answers and instrumental tools to cell biologists studying actin-based motility. The analysis of the entry process and in particular the studies derived from the specificity of internalin for its receptor will demonstrate how an apparently reductionist approach can help generating relevant animal models, identification of virulence factors and demonstration of their role in vivo. The virulence gene cluster and its regulation by PrfA will be presented and discussed in the framework of the recently determined genome sequence. PMID- 11890538 TI - Discovery of the anthrax toxin: the beginning of studies of virulence determinants regulated in vivo. AB - Anthrax kills many animal species. It was used to prove Koch's Postulates in 1876. Soon after that the classical bacterial toxins were produced in vitro, but up to 1950, a lethal toxin had not been demonstrated in either anthrax bacilli or filtrates from laboratory cultures. The cause of death had been an enigma for seventy years. During the 1950's, a toxin was recognized by examining bacteria and their products obtained from guinea pigs dying of anthrax. The toxin was found in their plasma and shown to contain two components. It was then produced in vitro and a third component recognized. The work reawakened interest in bacterial toxins after a period of dormancy and showed that toxins could be multicomponent. It demonstrated that previously unknown determinants of bacterial pathogenicity could be revealed by examining organisms grown in vivo. It was the beginning of such studies, which took a long time to evolve, but have now expanded greatly with the development of many new methods for examining bacterial activities in vivo. This paper is a personal account of the early work. PMID- 11890539 TI - Genomics of Neisseria meningitidis. AB - An important feature of disease caused by Neisseria meningitidis is the propensity to invade the meninges. Much progress has been made in our understanding of how this pathogen circumvents the physical properties of this cellular barrier. This review will address the new possibilities offered by the recent availability of meningococcal genome sequences. PMID- 11890540 TI - Role of the Hha/YmoA family of proteins in the thermoregulation of the expression of virulence factors. AB - Virulent bacteria are able to sense temperature changes and respond by modifying the expression of--among others--genes that code for virulence factors. The chromatin-associated protein H-NS has been shown to play a role in the thermomodulation of virulence factor expression. In addition to H-NS, proteins of the Hha/YmoA family have also been identified in different enterobacteria as participating in the thermoregulation of some virulence factors. For one of these proteins, the Hha protein, it has been shown that it interacts with H-NS, and both proteins form a nucleoid-protein complex responsible for the thermoregulation of, at least, E. coli hemolysin. The presence of genes coding for homologues of both proteins on some conjugative plasmids and their relation to thermoregulation suggests that this complex could also play a role in the regulation of plasmid transfer. PMID- 11890541 TI - Structure and function of membrane rafts. AB - Lipids do not always mix uniformly in membranes, but can cluster to form microdomains. We will consider one type of microdomain that can form in cell membranes. These are enriched in cholesterol and sphingolipids, and are referred to as rafts. Rafts probably exist in membranes in the liquid-ordered phase or a phase with similar properties. We will briefly review membrane lipid phase behavior, and the differences between liquid-crystalline, liquid-ordered, and gel phase membrane bilayer domains. We will present evidence suggesting that phospholipid-rich, liquid-crystalline phase domains and sphingolipid-rich, liquid ordered phase domains (rafts) can exist in equilibrium in biological membranes, especially the plasma membrane. Preferential partitioning of membrane proteins into rafts can affect function. Among the proteins that are targeted to rafts are those anchored in the outer leaflet of the membrane through covalent attachment to a special glycolipid, glycosyl phosphatidylinositol (GPI). Other proteins that are linked to saturated acyl chains, such as those that are directly acylated with two or more palmitate chains, or a palmitate and a myristate chain, are also targeted to rafts. Targeting of GPI-anchored proteins and other proteins to rafts plays a role in signal transduction in hematopoietic cells, and possibly also in sorting in intracellular membranes and regulation of cell-surface proteolysis in other mammalian cells. PMID- 11890542 TI - Detergent-resistant membrane microdomains and apical sorting of GPI-anchored proteins in polarized epithelial cells. AB - Detergent-insoluble microdomains or rafts play a crucial role in many cellular functions: membrane traffic, cell signalling and human diseases. In this work we investigate the role of rafts in the sorting of GPI-anchored proteins in polarized epithelial cells. In contrast to MDCK cells, the majority of endogenous GPI-anchored proteins are sorted to the basolateral surface of Fischer rat thyroid cells (Zurzolo et al., J. Cell Biol. 121, 1031-1039, 1993). We analyzed a set of transfected GPI proteins in order to understand the role of the GPI anchor and of association with rafts for apical sorting. We found that the GPI moiety is necessary but not sufficient for apical sorting of GPI proteins and that the ectodomain has a major role. We propose a new model in which the stabilization of proteins into rafts, probably mediated by interactions between protein ectodomains and a putative receptor, plays a crucial role in apical sorting. PMID- 11890543 TI - Lipid microdomains are involved in neurospecific binding and internalisation of clostridial neurotoxins. AB - The neuroparalytic syndromes of tetanus and botulism are caused by tetanus and botulinum neurotoxins, which are produced by bacteria of the genus Clostridia. These neurotoxins are structurally organised in three-domains endowed with different functions: specific interaction with the neuronal surface, membrane translocation and specific cleavage of three key components of the neurotransmitter release apparatus. Despite an identical intracellular activity, tetanus and botulinum neurotoxins are characterised by a differential intraneuronal trafficking, which is likely to be responsible for the different symptoms observed in clinical tetanus and botulism. This review aims to highlight recent discoveries on the recruitment of clostridial neurotoxins (CNTs) to the surface of neurons and neuronally-differentiated cell lines and to discuss their relevance for the internalisation and sorting of these neurotoxins. PMID- 11890544 TI - The Yersinia Ysc-Yop virulence apparatus. AB - The Yop virulon is an integrated system allowing extracellular Yersinia adhering at the surface of a target cell to inject an array of bacterial effectors into the eukaryotic cytosol. It consists of a type III secretion apparatus, called the Ysc injectisome and an array of proteins secreted by this apparatus, called Yops. The injectisome is made of about 25 Ysc proteins. The proximal part of the injectisome resembles the basal body of the flagellum while the most distal part is made of a secretin and a small needle protruding from the bacterial surface. Three of the Yops, namely YopB, YopD and LcrV, are required for the translocation of the others across the target cell membrane. They form some kind of a pore in the target cell membrane. Four Yop effectors, YopE, YopT, YpkA and YopH disturb the cytoskeleton dynamics by targeting monomeric GTPases of the Rho family. YopP downregulates the onset of the inflammatory response by blocking the NF-kappaB and MAPK pathways. Strong arguments indicate that it is a SUMO protease. Finally, YopM has been shown to travel to the nucleus of the target cell. PMID- 11890545 TI - The Dot/lcm transporter of Legionella pneumophila: a bacterial conductor of vesicle trafficking that orchestrates the establishment of a replicative organelle in eukaryotic hosts. AB - Legionella pneumophila are gram negative bacteria that parasitize professional phagocytes. When ingested by their eukaryotic hosts, these bacteria have the unique ability to alter maturation of the endocytic vacuoles in which they reside initially and create an organelle that supports replication. This review will focus on several scientific breakthroughs made recently that shed light on the mechanisms by which L. pneumophila are able to establish a niche permissive for intracellular growth. PMID- 11890546 TI - Mechanism of action of EPEC type III effector molecules. AB - Enteropathogenic E. coli (EPEC) is a prototypic member of the family of related 'attaching and effacing (A/E)' pathogens that induce diarrhoeal disease, especially to the young that can be fatal, of a wide range of mammalian species. Disease is correlated with the loss of absorptive gut epithelial microvilli and the reorganisation of host cytoskeletal proteins into pedestal-like structures beneath the adherent bacteria. These phenotypes are dependent on a pathogenicity island (LEE; Locus of Enterocyte Effacement) encoding a type III secretion system, secreted proteins, chaperone molecules, regulatory proteins and the bacterial outer membrane protein intimin. The type III secretion apparatus directs the transfer of specific proteins across the bacterial envelope, with a subset (EPEC secreted proteins - EspA, EspB and EspD) functioning to transfer effector proteins into host cells. These effector molecules subvert cellular processes that undoubtedly benefit the pathogen and contribute to disease. Three LEE-encoded EPEC effector molecules have so far been identified with one, Tir (Translocated intimin receptor), being transferred into host cells where it is modified by host kinases and becomes inserted into the plasma membrane to orchestrate cytoskeletal rearrangements linked to disease. This activity is dependent on its interaction with intimin and on tyrosine phosphorylation, with Tir-intimin interaction essential for virulence. A second effector Map, Mitochondrial-associated protein, is targeted to mitochondria where it has membrane-potential disrupting activity. The third, EspF disrupts intestinal barrier function and can induce host cell death by unknown mechanisms. Recent data relating to the mechanism by which Tir and Map function within host cells is discussed. PMID- 11890547 TI - Characterization of effector proteins translocated via the SPI1 type III secretion system of Salmonella typhimurium. AB - Salmonella spp. employ a conserved type III secretion system encoded within the pathogenicity island 1 (SPI1; centisome 63) to translocate effector proteins into the host cytosol. The translocated effector proteins trigger diverse responses including bacterial internalization. In a mutation analysis we have defined the set of effector proteins mediating tissue culture cell invasion. This set includes sopE2 (centisome 40-42), sopB (SPI5, centisome 20) and in the case of S. typhimurium SL1344 also the phage-encoded effector sopE (SopEphi, centisome 59 60). A triple mutant SL1344 derivative deficient of SopE, SopE2 and SopB was more than 100-fold attenuated in tissue culture cell invasion. Phylogenetic analyses indicate that the last common ancestor of all contemporary Salmonella lineages already harbored all genes necessary for host cell invasion, namely the SPI1 type III secretion system, sopE2 and sopB. SopE, which is 70% identical to sopE2 is only present in some Salmonella strains and emerged later well after the divergence of the contemporary Salmonella lineages. Interestingly, S. typhimurium strains that harbor sopE are associated with epidemics, arguing that sopE is one of the factors determining the "fitness" of a strain. We found that SopE can specifically activate a different set of host cellular RhoGTPases than SopE2. This allows the bacteria to fine tune host cellular responses very precisely and may offer an explanation for the improved epidemic fitness of sopE-positive S. typhimurium strains. PMID- 11890548 TI - Origin, originality, functions, subversions and molecular signalling of macropinocytosis. AB - Macropinocytosis refers to the formation of primary large endocytic vesicles of irregular size and shape, generated by actin-driven evaginations of the plasma membrane, whereby cells avidly incorporate extracellular fluid. Macropinosomes resemble "empty" phagosomes and show no difference with the "spacious phagosomes" triggered by the enteropathogenic bacteria Salmonella and Shigella. Macropinosomes may fuse with lysosomes or regurgitate their content back to the extracellular space. In multiple cell types, macropinocytosis is a transient response to growth factors. When amoebas are cultured under axenic conditions, macropinocytosis is induced so as to fulfil nutritional requirements. In immature dendritic cells, macropinocytosis allows for extensive sampling of soluble antigens; after a few days of maturation, this activity vanishes as processed peptides are being presented. Macropinosomes are also formed at the leading edge of motile leukocytes or neurons. In all these examples, macropinocytosis appears tightly regulated. Transformation of fibroblasts by Src or Ras also results in constitutive formation of macropinosomes at "ruffling" zones, that could be related to accelerated cell motility. Like phagocytosis, macropinocytosis depends on signalling to the actin cytoskeleton. We have explored this signalling in transformed cells. v-Src and K-Ras activate PI3K and PLC, as demonstrated by in situ production of the corresponding lipid products. Pharmacological inhibitors of PI3K and PLC and stable transfection leading to a dominant-negative PI3-kinase construct in transformed fibroblasts abolish macropinocytosis, demonstrating that both enzyme activities are essential. Conversely, stable transfection leading to a dominant-positive P13K in non-transformed fibroblasts is sufficient to induce macropinocytosis. Combination of experiments allows to conclude that P13K and PLC act in sequential order. In non-polarized cells expressing a thermosensitive v Src mutant, v-Src kinase activation accelerates fluid-phase endocytosis. In polarized MDCK cells, this stimulation occurs selectively at the apical domain and the response is selectively abrogated by pharmacological inhibitors of P13K and PLC. Thus, two paradigmatic oncogenes cause constitutive macropinocytosis. For v-Src, this response is polarized at the apical membrane. It is suggested that, in enterocytes that do not normally phagocytose, the P13K-PLC signalling pathway leading to selective induction of macropinocytosis at the luminal surface has been subverted by enteropathogenic bacteria to penetrate via "spacious phagosomes". PMID- 11890549 TI - Cytolethal distending toxins and activation of DNA damage-dependent checkpoint responses. AB - Cytolethal distending toxins (CDTs) are unique among bacterial protein toxins in their ability to cause DNA damage, due to their functional similarity to the mammalian deoxyribonuclease I (DNase I). The cellular response to CDT intoxication is characterised by activation of DNA damage-induced checkpoint responses, and the final outcome is cell type dependent. Cells of epithelial origin and normal keratinocytes are arrested in the G2 phase of the cell cycle, normal fibroblasts are also arrested in G1, while B cells die of apoptosis. CDTs are encoded by three linked genes (cdtA, cdtB and cdtC), and CdtB is the toxin subunit which possesses the DNase I-like activity. All the three genes have to be present in the bacterium in order to produce an active cytotoxin, however cytotoxic Haemophilus ducreyi CDT, purified from a CdtABC recombinant E. coli strain, contains the CdtB and CdtC subunits, suggesting that they constitute the holotoxin and that CdtC may be required for CdtB internalization. The role of the CdtA subunit is currently unknown, but it might modify and therefore activate CdtC. This review will focus on the cellular responses induced by CDTs in mammalian cells. PMID- 11890550 TI - Resistance to phagocytosis by Yersinia. AB - Enteropathogenic species of the genus Yersinia penetrate the intestinal epithelium and then spread to the lymphatic system, where they proliferate extracellularly. At this location, most other bacteria are effectively ingested and destroyed by the resident phagocytes. Yersinia, on the other hand binds to receptors on the external surface of phagocytes, and from this location it blocks the capacity of these cells to exert their phagocytic function via different receptors. The mechanism behind the resistance to phagocytosis involves the essential virulence factor YopH, a protein tyrosine phosphatase that is translocated into interacting target cells via a type III secretion machinery. YopH disrupts peripheral focal complexes of host cells, seen as a rounding up of infected cells. The focal complex proteins that are dephosphorylated by YopH are focal adhesion kinase and Crk-associated substrate, the latter of which is a common substrate in both professional and non-professional phagocytes. In macrophages additional substrates have been found, the Fyn-binding/SLP-76 associated protein and SKAP-HOM. Phagocytosis is a rapid process that is activated when the bacterium interacts with the phagocyte. Consequently, the effect exerted by a microbe to block this process has to be rapid and precise. This review deals with the mechanisms involved in impeding uptake as well as with the role of the YopH substrates and focal complex structures in normal cell function. PMID- 11890551 TI - Toxin-induced calcium oscillations: a novel strategy to affect gene regulation in target cells. AB - Although the mucosal linings continuously are exposed to microbes, the microbes rarely induce disease. This is because mucosal surfaces are protected by a first line of host defence termed the innate immunity system. The innate immune response is an outcome of the complex interplay between microbes and host target cells, and leads to the activation of inflammatory processes. Although inflammation is essential for clearing out infectious agents, it can also be harmful to the host and is therefore subjected to control at multiple levels. We recently discovered that alpha-haemolysin, a toxin secreted by uropathogenic E. coli induces constant, low-frequency Ca2+ oscillations in renal epithelial cells (Uhlen et al., Nature 405, 694-696 (2000)). Ca2+ oscillation at a specific periodicity of 12 min was found to affect gene expression in target epithelial cells, as the proinflammatory cytokine IL-6 and chemokine IL-8 were specifically induced by alpha-haemolysin-induced Ca2+ oscillations. This demonstrates a novel feature of bacterial toxin effects on host target cells: as inducers of second messenger responses which fine-tune gene expression in target epithelial cells into pathways leading to e. g. a pro-inflammatory response. PMID- 11890552 TI - Retrograde transport of ricin. AB - The plant toxin ricin binds to terminal galactose-containing cell-surface receptors. The toxin is endocytosed and transported to the Golgi apparatus. Recent evidence suggests that ricin binds to galactosylated calreticulin, which may carry the toxin from the Golgi apparatus to the endoplasmic reticulum (ER). From the ER, the ricin A fragment is translocated to the cytosol. Ricin is perceived to be a candidate for ER-associated degradation (ERAD) and is translocated through the Sec61p translocon to the cytosol. Part of the toxin is degraded by the proteasome, but a fraction of the ricin avoids degradation and inhibits protein synthesis by inactivating ribosomes, ultimately leading to cell death. PMID- 11890554 TI - New insights into the structure-function relationships and therapeutic applications of cholera-like enterotoxins. AB - Cholera toxin and E. coli heat-labile enterotoxin are structurally homologous proteins comprised of an enzymatically active A-subunit and five B-subunits that bind with high affinity to GM1-ganglioside receptors found on the surface of mammalian cells. The B-subunits have long been thought of simply as trafficking vehicles that trigger entry and subsequent delivery of the 'toxic' A-subunit into cells. Indeed, such is the capacity of the B-subunits to enter cells, that they have been developed as generic carriers for attachment and delivery of a variety of peptides into mammalian cells. However, the B-subunits also appear to possess discrete 'signalling functions', that induce both transcription factor and cell activation. These are thought to be directly responsible for the potent immunomodulatory properties of the B-subunits, and have resulted in their use as adjuvants and as agents to suppress inflammatory immune disorders. The relationship between the signalling properties of the B-subunits and their capacity to act as trafficking vehicles has remained unclear. In an effort to understand the structural requirements for these two functions, a set of mutant B subunits, with amino acid substitutions at position His-57, have been generated and studied. Importantly, such mutant B-subunits retain an ability to bind with high affinity to GM1 and to traffic into cells, but have entirely lost their capacity to activate immune cell populations. Thus, while binding via GM1 appears to be sufficient to trigger cellular uptake it is not sufficient to activate signal transduction. The His-57 region is therefore speculated to be actively engaged in triggering signalling events, possibly via cognate interaction with other cell surface molecules. PMID- 11890553 TI - The ARTT motif and a unified structural understanding of substrate recognition in ADP-ribosylating bacterial toxins and eukaryotic ADP-ribosyltransferases. AB - ADP-ribosylation is a widely occurring and biologically critical covalent chemical modification process in pathogenic mechanisms, intracellular signaling systems, DNA repair, and cell division. The reaction is catalyzed by ADP ribosyltransferases, which transfer the ADP-ribose moiety of NAD to a target protein with nicotinamide release. A family of bacterial toxins and eukaryotic enzymes has been termed the mono-ADP-ribosyltransferases, in distinction to the poly-ADP-ribosyltransferases, which catalyze the addition of multiple ADP-ribose groups to the carboxyl terminus of eukaryotic nucleoproteins. Despite the limited primary sequence homology among the different ADP-ribosyltransferases, a central cleft bearing the NAD-binding pocket formed by the two perpendicular beta-sheet cores has been remarkably conserved between bacterial toxins and eukaryotic mono- and poly-ADP-ribosyltransferases. The majority of bacterial toxins and eukaryotic mono-ADP-ribosyltransferases are characterized by conserved His and catalytic Glu residues. In contrast, diphtheria toxin, Pseudomonas exotoxin A, and eukaryotic poly-ADP-ribosytransferases are characterized by conserved Arg and catalytic Glu residues. Structural and mutagenic studies of the NAD-binding core of a binary toxin and a C3-like toxin identified an ARTT motif (ADP-ribosylating turn-turn motif) that is implicated in substrate specificity and recognition. Here we apply structure-based sequence alignment and comparative structural analyses of all known structures of ADP-ribosyltransfeases to suggest that this ARTT motif is functionally important in many ADP-ribosylating enzymes that bear a NAD-binding cleft as characterized by conserved Arg and catalytic Glu residues. Overall, structure-based sequence analysis reveals common core structures and conserved active sites of ADP-ribosyltransferases to support similar NAD-binding mechanisms but differing mechanisms of target protein binding via sequence variations within the ARTT motif structural framework. Thus, we propose here that the ARTT motif represents an experimentally testable general recognition motif region for many ADP-ribosyltransferases and thereby potentially provides a unified structural understanding of substrate recognition in ADP-ribosylation processes. PMID- 11890555 TI - Molecular interactions of the CcdB poison with its bacterial target, the DNA gyrase. AB - The ccd poison/antidote system of the F plasmid encodes CcdB, a toxin targeting the essential DNA gyrase of E. coli, and CcdA, the unstable antidote that interacts with CcdB to neutralise its toxicity. Gyrase belongs to the topoisomerase II class of enzymes and is a well-validated target for efficient therapeutic drugs, i. e. the quinolones. CcdB acts on gyrase in a similar way as quinolones do, both compounds induce double-strand breaks in DNA. Interestingly, the CcdB-binding domain of gyrase is different than that of quinolones. Therefore, novel classes of therapeutic drugs could be derived from the analysis of the interaction between CcdB and gyrase. PMID- 11890557 TI - Rho-activating Escherichia coli cytotoxic necrotizing factor 1: macropinocytosis of apoptotic bodies in human epithelial cells. AB - Some pathogenic Escherichia coli strains produce a protein toxin, named cytotoxic necrotizing factor 1 (CNF1), which permanently activates proteins belonging to the Rho family. In epithelial cells, the consequence of this activation is the rearrangement of the actin cytoskeleton and the promotion of an intense and generalized ruffling activity. This leads, in turn, to the induction of a phagocytic-like behavior called macropinocytosis that, in the case of CNF1, depends on the coordinate activation of Rho, Rac and Cdc42. Following internalization, the ingested material is discharged into Rab-7 and Lamp-1 positive acidic vesicles where it probably undergoes degradation. By exerting this activity, CNF1-activated epithelial cells might support the scavenging activity of macrophages during bacterial overgrowth. PMID- 11890556 TI - The neutrophil-activating protein of Helicobacter pylori. AB - Infection of the stomach mucosa by the gastric pathogen Helicobacter pylori is accompanied by a large infiltration of neutrophils and monocytes which are believed to contribute substantially to H. pylori-induced gastritis. A protein was identified (HP-NAP for neutrophil-activating protein from H. pylori) that was capable of increasing the adhesion of neutrophils to endothelial cells. We have demonstrated that HP-NAP is a dodecamer composed of identical 17-kDa subunits that induces the production of reactive oxygen radicals (ROIs) by neutrophils via a cascade of intracellular activation events. HP-NAP has also been shown to be chemotactic for neutrophils and monocytes, and a majority of H. pylori-infected patients have been found to produce antibodies specific for HP-NAP making it a strong vaccine candidate. More recently it has been shown that HP-NAP can stimulate tissue factor and plasminogen activator inhibitor-2 production by human monocytes. While structurally similar to the Escherichia coli DNA-binding protein Dps, HP-NAP has characteristics that are more similar to bacterioferritins being capable of binding up to 500 atoms of iron in vitro. Further study, however, has revealed that synthesis of HP-NAP in H. pylori is not altered by the addition or subtraction of metal ions from its growth medium suggesting that the primary role of the protein in vivo is not as a metal-binding protein. A number of other reports have proposed that HP-NAP acts as an adhesin being capable of binding several different compounds in vitro. Sequence analysis of the genomes of several other bacteria reveal that many possess Dps/HP-NAP-like proteins. The preliminary characterisation of some of these proteins will be discussed. PMID- 11890558 TI - Bacterial persistence within erythrocytes: a unique pathogenic strategy of Bartonella spp. AB - The genus Bartonella comprises human-specific and zoonotic pathogens responsible for a wide range of clinical manifestations, including Carrion's disease, trench fever, cat scratch disease, bacillary angiomatosis and peliosis, endocarditis and bacteremia. These arthropod-borne pathogens typically parasitise erythrocytes in their mammalian reservoir host(s), resulting in a long-lasting haemotropic infection. We have studied the process of Bartonella erythrocyte parasitism by tracking green fluorescent protein-expressing bacteria in the blood of experimentally infected animals. Following intravenous infection, bacteria colonise a yet enigmatic primary niche, from where they are seeded into the blood stream in regular intervals of approximately five days. Bacteria invade mature erythrocytes, replicate temporarily and persist in this unique intracellular niche for the remaining life span of the infected erythrocytes. A triggered antibody response typically results in an abrogation of bacteremia within 3 months of infection, likely by blocking new waves of bacterial invasion into erythrocytes. The recent establishment of genetic tools for Bartonella spp. permitted us to identify several putative pathogenicity determinants. Application of differential fluorescence induction technology resulted in the isolation of bacterial genes differentially expressed during infection in vitro and in vivo, including an unknown family of autotransporter proteins as well as a novel type IV secretion system homologous to the conjugation system of E. coli plasmid R388. Mutational analysis of a previously described type IV secretion system displaying homology to the virB locus of Agrobacterium tumefaciens provided the first example of an essential pathogenicity locus in Bartonella. Though required for establishing haemotropic infection, it remains to be demonstrated if this type IV secretion system is necessary for colonisation of the primary niche or for the subsequent colonisation of erythrocytes. PMID- 11890559 TI - Delivery of protein antigens and DNA by attenuated intracellular bacteria. AB - On the basis of attenuated intracellular bacteria, we have developed two delivery systems for either heterologous proteins or DNA vaccine vectors. The first system utilizes attenuated strains of Gram-negative bacteria which are engineered to secrete heterologous antigens via the alpha-hemolysin secretion system (type I) of Escherichia coli. The second system is based on attenuated suicide strains of Listeria monocytogenes, which are used for the direct delivery of eukaryotic antigen expression vectors into professional antigen-presenting cells (APC) like macrophages and dendritic cells in vitro and can be also used in animal models. PMID- 11890560 TI - Pharmacology of antidepressants: focus on nefazodone. AB - The past decade has witnessed the introduction of a diverse group of antidepressants from a variety of distinct chemical classes, each with their own specificity for neurochemical transmitters, receptors, and cytochrome P450 isozymes. This review focuses on nefazodone, a distinct antidepressant with efficacy for the treatment of depression with depression-related anxiety symptoms, an established tolerability profile, and a multimodal mechanism of action. Relevant pharmacologic and pharmacodynamic effects are summarized that support nefazodone as an attractive choice for both the short- and long-term treatment of depression. PMID- 11890561 TI - Clinical use of nefazodone in major depression: a 6-year perspective. PMID- 11890562 TI - Overview of psychiatric disorders and the role of newer antidepressants. PMID- 11890563 TI - Six-year perspectives on the safety and tolerability of nefazodone. PMID- 11890564 TI - Antidepressant dosing and switching guidelines: focus on nefazodone. AB - Dosing strategies form a fulcrum between patient-related and provider-related dimensions of treatment. The former focus on perceptions of efficacy and safety and adherence to the regimen, whereas the latter focus on symptom identification and appropriate drug selection. Appropriate dosing strategies may modulate adverse effects, allowing the patient to move more comfortably toward an efficacious response. Dosing also is an important dimension in switching between 2 antidepressants when the efficacy of the first agent is suboptimal. Nefazodone is effective for the management of acute, severe, and chronic major depression and relapse prevention. Response rates with nefazodone are comparable to those of imipramine and most SSRIs. In clinical trials, the efficacy of nefazodone was most clearly established at doses between 300 and 600 mg/day. At this dose range, discontinuations because of adverse events are low. PMID- 11890565 TI - Cost savings with nefazodone in treating depression. AB - Pharmacoeconomic analysis of antidepressant therapy is an important tool for ensuring the most cost-cognizant approach to treat a particular mental disorder. As the number of effective antidepressant compounds continues to grow, the drug selection process must consider not only the cost of the drug itself, but also costs associated with treatment failure and management of untoward and unexpected side effects. In economic studies conducted in North America and England using a decision analysis model and a direct annual cost model, nefazodone has been shown to have an impact on costs associated with depression when compared with imipramine and fluoxetine. Nefazodone also can reduce depression-related anxiety and agitation symptoms early in treatment, and, because it improves subjective and objective sleep measures, use of concomitant anxiolytics or sedative hypnotics with nefazodone has been shown to be less frequent and less costly than with selective serotonin reuptake inhibitors. PMID- 11890566 TI - Antibiotic susceptibility of blood culture isolates of Enterobacteriaceae. A Norwegian multicenter study. AB - From May to November 1997 each of six major hospitals throughout Norway collected 72 to 104 consecutive blood culture isolates of Enterobacteriaceae, altogether 563 isolates. Escherichia coli was the predominating organism (69%), followed by Klebsiella spp. (15%), Enterobacter spp. (6%), and Proteus mirabilis (4%). The susceptibility of the isolates to ampicillin, cefuroxime, ceftazidime, imipenem, tobramycin, and ciprofloxacin was determined by the E-test. 37% and 7% of the isolates were resistant to ampicillin and cefuroxime, respectively, and 1% were resistant to ceftazidime and tobramycin. Only one isolate of P. mirabilis was imipenem resistant. All isolates were susceptible to ciprofloxacin. The prevalence of ampicillin-resistant isolates at each hospital varied from 21 to 45%, and of cefuroxime-resistant isolates from 3 to 9%. The results were compared with those of a similar study performed in 1991-1992. No significant changes in the susceptibility to the various agents could be demonstrated. The high frequency of isolates resistant to ampicillin has clearly limited the usefulness of this agent in the treatment of septicemia and other serious infections caused by Enterobacteriaceae. PMID- 11890567 TI - Production and partial characterization of mouse monoclonal antibodies recognizing common cytokine receptor gamma chain (gammac) of human, mouse and primate origin. AB - Monoclonal antibodies specific for the common cytokine receptor gamma chain, gammac, were produced using traditional hybridoma technology. Fusion of P3X63 Ag8.653 myeloma cells with splenocytes from Balb/c mice immunized with Spodoptera frugiperda insect cells infected with the recombinant baculovirus VL1392-hIL 2Rgamma resulted in several hybridoma cell clones producing monoclonal gammac specific antibodies. Four of these antibody-producing clones, IIIC3, IIIE8, IG3 and IF10C5, were further characterized by immunoblotting, flow cytometry and ELISA. Data are presented demonstrating that the generated monoclonal antibodies can identify the extracellular domain of the common cytokine receptor gamma chain of human and mouse origin, and two of the antibodies recognize gammac of primate origin as well. PMID- 11890568 TI - In situ hybridisation for identification and differentiation of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae and Mycoplasma hyorhinis in formalin-fixed porcine tissue sections. AB - Oligonucleotide probes targeting 16S ribosomal RNA were designed for species specific identification of the porcine mycoplasmas Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma hyosynoviae using a fluorescent in situ hybridisation assay. The specificity of the probes was evaluated using pure cultures as well as porcine tissue sections with artificial presence of mycoplasma, and the probes were found specific for the target organisms. The assay was applied on sections of 28 tissue samples from pigs infected with one or more of the three Mycoplasma species as determined by cultivation. M. hyopneumoniae and M. hyorhinis were identified in accordance with cultivation in lung sections from nine pigs affected by catarrhal to purulent bronchopneumonia. Likewise, in eight cases of fibrinous pericarditis, M. hyopneumoniae, M. hyorhinis and M. hyosynoviae were the infectious agents according to cultivation and were correctly identified by in situ hybridisation. Out of 11 joints cultivation positive for M. hyosynoviae, the probe was only able to identify M. hyosynoviae in eight cases probably due to a low number of microorganisms in the tissue sections. The in situ hybridisation assay is well suited for use in diagnostic and experimental work as well as a tool for pathogenesis studies. PMID- 11890569 TI - An enzyme-based in situ hybridisation method for the identification of Streptococcus suis. AB - A method for enzyme-based in situ hybridisation of Streptococcus suis was developed. It enables the light microscopic localization of bacterial ribosomal RNA (rRNA) in formalin-fixed paraffin-embedded tissues. A unique sequence in the 16S rRNA of S. suis was targeted. Different pretreatment protocols were applied to facilitate probe penetration and multiple detection systems were tested. The results were compared to those obtained by immunohistochemistry. Pretreatment was necessary to obtain a signal by in situ hybridisation. The use of proteinase-K pretreatment was optimal regarding sensitivity and preservation of tissue morphology. A strong specific in situ hybridisation signal was achieved in tissue sections containing S. suis in microcolonies and the microanatomy of the surrounding tissue was easily assessed. However, the signal distribution differed from that found immunohistochemically and low-grade infection could not be detected by in situ hybridisation. These findings were interpreted as reflecting the physiological state of the bacteria. Thus, this method could prove useful in future studies of the infection pathogenesis. PMID- 11890570 TI - Killing of Gram-positive cocci by fatty acids and monoglycerides. AB - The susceptibilities of three Gram-positive cocci to medium-chain saturated and long-chain unsaturated fatty acids and their one-monoglycerides were studied. The bacteria were incubated with equal volumes of lipid solutions for 10 min. Lauric acid, palmitoleic acid and monocaprin reduced the number of CFU by 6.0 log10 or greater at 5 mM concentration for streptococci of group A (GAS) and group B (GBS). When further compared at lower concentrations and after longer incubation time monocaprin proved to be the most active. Capric acid showed the highest activity against Staphylococcus aureus at 10 mM. However, at lower concentrations monocaprin was the only lipid that showed significant activity against S. aureus. The mode of action of monocaprin against GBS was studied by a novel two-color fluorescent assay of bacterial viability and by electron microscopy. The results indicate that the bacteria are killed by disintegration of the cell membrane by the lipid, leaving the bacterial cell wall intact. The highly lethal effect of monocaprin indicates that this lipid might be useful as a microbicidal agent for prevention and treatment of infections caused by these bacteria. PMID- 11890571 TI - Apoptotic cell death induced by Acinetobacter baumannii in epithelial cells through caspase-3 activation. AB - Epithelial cell death induced by Acinetobacter baumannii infection was investigated using in vitro assays. Eight hours after live A. baumannii infection, HeLa cells exhibited detachment from the dish, rounding morphologies, high proportions of trypan blue-positive cells and extensive DNA breakdown with faint apoptotic banding, which is indicative of cells undergoing apoptosis. The enzymatic activity of caspase-3 was increased in cells as early as 2 h after infection. In addition, apoptosis of HeLa cells was induced by treatment with bacterial culture filtrates but not with formalin-killed bacteria. These results indicate that A. baumannii infection triggers apoptosis in HeLa cells through caspase-3 activation. PMID- 11890572 TI - Factors affecting the cytology outcome of Pap smears--a brief approach to internal quality control in private cytopathology laboratory practice. AB - We evaluated the effect of intrauterine device (IUD), patient age and hormone replacement therapy (HRT) on cytology outcome in Pap smears together with the important IQC procedures: 1) manual double-screening by cytotechnologists, and 2) retrospective senior pathologist review during 1996-1999. The results from primary double-screening (119 of 87,409 Pap smears) showed an excellent inter observer correlation. The estimation of hormonal effects showed higher incidence of disagreements (p=0.013) in patients <47 yr. Some individual trends were found in the assessments of both cellular atypia (p=0.012) and Papanicolaou classification (p=0.018). The IUD had no influence on the accuracy when the degree of inflammatory reaction was evaluated (p>0.050), but showed an adverse effect on the estimation of cellular atypia (p=0.001). HRT distinctly equalized the entire sample material, since fewer disagreements were found in the age groups <47 yr and >47 yr when estimating the hormonal effects (p=0.013), inflammatory reaction (p=0.044) or cellular atypia (p=0.006) compared to those without HRT. The continuous cytopathologist supervision had a positive impact on the accuracy of hormonal effect estimation during the 4 years. The senior cytopathologists' reviews (354 of 87,409 Pap smears) showed mutually good interobserver correlation, and diagnostic conclusions of the same specimens differed only slightly between the cytopathologists. We found these state-of-the art cytopathological IQC procedures to be effective and fit-for-purpose when evaluating hormonal effects, inflammatory reaction and cellular atypia. PMID- 11890573 TI - Poorly differentiated endometrial sarcoma with clear cell features. AB - A case of poorly differentiated endometrial sarcoma with clear cell features in a 46-year-old female is presented. A 9 x 8 x 6 cm, transmural, white solid tumor in the uterine fundic wall was characterized histologically by infiltrative growth and a solid arrangement of tumor cells with clear cytoplasm. The tumor showed vascular invasion, moderate to severe cellular atypia, mitotic activity of 20 per 10 high-power fields and necrosis. It was immunohistochemically positive for vimentin and negative for CAM5.2, epithelial membrane antigen, desmin, alpha smooth muscle actin, muscle actin, estrogen and progesterone receptors. Electron microscopy showed that the clear cytoplasm was attributed to the presence of numerous dilated mitochondria and lipid droplets. Flow cytometry showed a diploid phenotype. Pathologists should be aware of the existence of this type of endometrial sarcoma with clear cells and should not confuse it with other neoplasma showing clear cell features. PMID- 11890575 TI - Analysis of Borrelia burgdorferi sensu lato isolated from cerebrospinal fluid. AB - Involvement of the nervous system in Lyme borreliosis may occur with or without erythema migrans and it may present with a variety of neurological symptoms. In this study we analysed phenotypic and genotypic characteristics of 40 Borrelia strains isolated from cerebrospinal fluid (CSF) of 38 Slovenian patients with different clinical manifestations of Lyme borreliosis. In seven of the patients, Borreliae were also isolated from skin lesions. Species identification and plasmid profiles were determined by pulsed-field gel electrophoresis and protein profiles by SDS-PAGE. MluI digestion profiles of Borrelia burgdorferi sensu lato DNA showed that 25 (62.5%) isolates were B. garinii, 14 (35%) B. afzelii, and one (2.5%) B. burgdorferi sensu stricto. All strains, except one, possessed a large plasmid and a varying number of smaller plasmids. Three (7.5%) isolates exhibited an unusual plasmid profile, with a large plasmid dimer or three copies of the large plasmid. In protein analyses, all strains expressed OspA protein. OspB was present significantly more often in B. afzelii than B. garinii strains (p=0.0000), while OspC was more often present in B. garinii than B. afzelii strains (p=0.0052). In the seven patients with Borreliae isolated also from the skin, the CSF and skin isolates were identical, either B. garinii (six patients) or B. afzelii (one patient). Species and plasmid heterogeneity as well as antigen diversity could play a role in the pathogenesis of the infection. When combined with our own earlier data, the results suggest species-related organotropism. PMID- 11890574 TI - Angioimmunoblastic T-cell lymphoma with hyperplastic germinal centers: a clinicopathological and immunohistochemical study of 10 cases. AB - An absence of germinal centers is one of the histological characteristics of angioimmunoblastic T-cell lymphoma (AITL). We report here 10 unusual cases of AITL with hyperplastic germinal centers. The clinical presentation of each patient was characterized by generalized lymphadenopathy, constitutional symptoms and polyclonal hypergammaglobulinemia. The initial biopsy findings of each patient were similar and were characterized by hyperplastic germinal centers with ill-defined borders and a proliferation of high endothelial venules (HEV). In the paracortical area there was a mixed infiltrate including irregularly shaped clusters or small nests of clear cells in all cases. Moreover, the clear cells invaded the lymphoid follicles, resulting in expansion of the germinal centers, except for one case. Immunohistochemistry revealed that the tumor cells, including clear cells, were CD4-expressing T cells. Some of the atypical lymphocytes were also Bcl-6-positive. A majority of the follicular dendritic cell networks showed a normal/reactive or an expanded/disrupted pattern in all cases. Moreover, three lesions possessed a few large irregularly shaped proliferations of follicular dendritic cells around the HEV Four cases progressed to AITL within a few years. The present 10 cases probably represent an early stage of AITL preceding follicular dendritic cell hyperplasia. Detection of clear cells, Bcl-6 positive atypical T lymphocytes, and foci of irregularly shaped proliferation of follicular dendritic cells appears to be critical for early diagnosis and treatment of AITL with hyperplastic follicles. PMID- 11890576 TI - Postmitotic basal cells in squamous cell epithelia are identified with Dolichos biflorus agglutinin--functional consequences. AB - Dolichos biflorus agglutinin (DBA) is a plant lectin specifically recognizing alpha-N-acetylgalactosamine. Controversial reports regarding the binding of DBA to the epidermis have been published. Using a double labeling procedure at the single-cell level, we studied the expression of DBA-reactive binding sites in conjunction with markers of cell proliferation and differentiation in normal human epidermis, cornea, and malignant tumors as well as in cultured keratinocytes. The results characterize the cells recognized by DBA as postmitotic early differentiating cells, identifiable by their lack of expression of the proliferation marker (Ki-67). The Golgi complex of a limited number of cultured keratinocytes was recognized by DBA and some of these cells show the accumulation of beta1 integrin chain in the Golgi complex. This process seems to be important for the migration of postmitotic cells from the basal to the suprabasal layers. PMID- 11890577 TI - Moderate physical activity is associated with higher bone mineral density in postmenopausal women. AB - OBJECTIVES: To determine the associations between different levels of habitual physical activity, hormone replacement therapy (HRT), and bone mineral density (BMD) in postmenopausal women. DESIGN: Cross-sectional. SETTING: Academic medical center. PARTICIPANTS: Twenty sedentary women, 20 active nonathletic women, and 23 endurance-trained athletes, all of whom were postmenopausal, with half of each group on and half not on HRT. MEASUREMENTS: BMD and body composition determined by dual energy x-ray absorptiometry, maximal oxygen consumption (VO2max), dietary history by questionnaire, and vitamin D receptor (VDR) genotyping on deoxyribonucleic acid. RESULTS: Body weight was higher in the active nonathletic than in the sedentary and athletic women. Body fat was lower and VO2max higher in the athletic women than in the sedentary and the active nonathletic women. Physical activity level was significantly associated with BMD in three of the five measurements taken (L1-L4 lumbar spine, trochanter, total body; all P < .05). These differences were also generally significant after adjusting for body weight. The association between physical activity status and BMD at the neck of the femur and Ward's triangle bordered on significance (P = .07-.09). At most sites, the active nonathletic women had higher BMD than did the sedentary and athletic women. HRT was significantly associated only with total body BMD (P < .05). The groups were similar in terms of dietary habits (protein, calcium, sodium, phosphorus intake); VDR genotypes; and family, smoking, and nutritional histories. CONCLUSION: Given the similarity of the groups with respect to other factors that affect BMD, it appears that prolonged low-to-moderate-intensity physical activity, but not the same number of years of higher-intensity training for competitive events, was independently associated with higher BMD. PMID- 11890578 TI - Task-specific resistance training to improve the ability of activities of daily living-impaired older adults to rise from a bed and from a chair. AB - OBJECTIVES: To determine the effect of a 12-week intervention to improve the ability of disabled older adults to rise from a bed and from a chair. DESIGN: Subjects were randomly allocated to either a 12-week task-specific resistance training intervention (training in bed- and chair-rise subtasks, such as sliding forward to the edge of a chair with the addition of weights) or a control flexibility intervention. SETTING: Seven congregate housing facilities. PARTICIPANTS: Congregate housing residents age 65 and older (n = 161, mean age 82) who reported requiring assistance (such as from a person, equipment, or device) in performing at least one of the following mobility-related activities of daily living: transferring, walking, bathing, and toileting. MEASUREMENTS: At baseline, 6 weeks, and 12 weeks, subjects performed a series of bed- and chair rise tasks where the rise task demand varied according to height of the head of the bed, chair seat height, and use of hands. Outcomes were able or unable to rise and, if able, the time taken to rise. Logistic regression for repeated measures was used to test for differences between tasks in the ability to rise. Following log transformation of rise time, a linear effects model was used to compare rise time between tasks. RESULTS: Regarding the maximum total number of bed- and chair-rise tasks that could be successfully completed, a significant training effect was seen at 12 weeks (P = .03); the training effect decreased as the total number of tasks increased. No statistically significant training effects were noted for rise ability according to individual tasks. Bed- and chair rise time showed a significant training effect for each rise task, with analytic models suggesting a range of approximately 11% to 20% rise-time (up to 1.5 seconds) improvement in the training group over controls. Training effects were also noted in musculoskeletal capacities, particularly in trunk range of motion, strength, and balance. CONCLUSIONS: Task-specific resistance training increased the overall ability and decreased the rise time required to perform a series of bed- and chair-rise tasks. The actual rise-time improvement was clinically small but may be useful over the long term. Future studies might consider adapting this exercise program and the focus on trunk function to a frailer cohort, such as in rehabilitation settings. In these settings, the less challenging rise tasks (such as rising from an elevated chair) and the ability to perform intermediate tasks (such as hip bridging) may become important intermediate rehabilitation goals. PMID- 11890579 TI - Muscle size responses to strength training in young and older men and women. AB - OBJECTIVES: To examine the possible influences of age and gender on muscle volume responses to strength training (ST). DESIGN: Prospective intervention study. SETTING: University of Maryland Exercise Science and Wellness Research Laboratories. PARTICIPANTS: Eight young men (age 20-30 years), six young women (age 20-30 years), nine older men (age 65-75 years), and ten older women (age 65 75 years). INTERVENTION: A 6-month whole-body ST program that exercised all major muscle groups of the upper and lower body 3 days/week. MEASUREMENTS: Thigh and quadriceps muscle volumes and mid-thigh muscle cross-sectional area (CSA) were assessed by magnetic resonance imaging before and after the ST program. RESULTS: Thigh and quadriceps muscle volume increased significantly in all age and gender groups as a result of ST (P < .001), with no significant differences between the groups. Modest correlations were observed between both the change in quadriceps versus the change in total thigh muscle volume (r = 0.65; P < .001) and the change in thigh muscle volume versus the change in mid-thigh CSA (r = 0.76, P < .001). CONCLUSIONS: The results indicate that neither age nor gender affects muscle volume response to whole-body ST. Muscle volume, rather than muscle CSA, is recommended for studying muscle mass responses to ST. PMID- 11890580 TI - Lipoprotein metabolism in Japanese centenarians: effects of apolipoprotein E polymorphism and nutritional status. AB - OBJECTIVES: To assess the complex interaction of apolipoprotein (apo) E polymorphisms and environmental factors on lipoprotein profile in centenarians. DESIGN: Cross-sectional analysis. SETTING: Tokyo metropolitan area. PARTICIPANTS: Seventy-five centenarians and 73 healthy older volunteers (mean age 63.1 +/- 10.0) living in the Tokyo metropolitan area. MEASUREMENTS: Plasma lipids and lipoproteins, cholesteryl ester transfer protein mass, apo E phenotype, body mass index, nutritional indices (serum albumin, prealbumin, transferrin), dietary intake, inflammation markers (C-reactive protein (CRP), interleukin-6 (IL-6)), activities of daily living, and cognitive function. RESULTS: In comparison with older people, the centenarians had low concentrations of total and low-density lipoprotein cholesterol (LDL-C) and a relative predominance of high-density lipoprotein 2 cholesterol. No environmental factor, except the number of apo E epsilon2 alleles, was a significant determinant of LDL-C and apo B, suggesting that the low apo B-containing lipoprotein in centenarians may be attributable to a genetic cause. Centenarians had elevated levels of lipoprotein (a) and decreased high-density lipoprotein cholesterol (HDL-C), which seem to be an unfavorable lipoprotein profile. Lower levels of HDL-C in the centenarians were associated with decreased serum albumin, elevated CRP and IL-6 levels, and cognitive impairment, suggesting that HDL-C could be a sensitive marker for frailty and comorbidity in the oldest old. CONCLUSIONS: Low levels of apo B containing lipoproteins attributable to a genetic cause may be advantageous for longevity. Lipoprotein profiles in centenarians were consistently related to the subjects' nutritional status, inflammation markers, and apo E polymorphisms. The results provide evidence for the importance of maintaining nutritional status in the very old. PMID- 11890581 TI - Preventing falls in older people: impact of an intervention to reduce environmental hazards in the home. AB - OBJECTIVES: To evaluate the impact of an intervention to reduce fall hazards in the homes of older people. DESIGN: The intervention was administered to the 570 subjects in the experimental arm of a randomized controlled trial, with follow-up of subjects for 1 year. SETTING: Community-based seniors living in Perth, Australia. PARTICIPANTS: People age 70 and older. INTERVENTION: Registered nurses delivered the intervention. It consisted of a home hazard assessment, an educational strategy on general fall hazard reduction and ways to reduce identified home hazards, and the free installation of safety devices: grab rails, nonslip stripping on steps, and double-sided tape for floor rugs and mats. All intervention subjects received the home hazard assessment, and 96% received the educational strategy. Grab rails were installed in 77% of homes, rugs were stabilized in 8%, and nonslip step stripping was installed in 36%. MEASUREMENTS: Hazard prevalence was assessed at baseline in all homes and 11 months later in a random sample of 51 homes. Action taken in response to the intervention was assessed by a self-completed postal questionnaire completed 11 months after the intervention. RESULTS: All homes had at least one fall hazard. The most prevalent were floor rugs and mats (mean of 14 per home), stepovers (Stepovers are structural changes to the height of the floor that were designed to be stepped over rather than stepped upon, for example, the lip of a shower or a bath side.) (mean of seven per home), steps (mean of four per home), and trailing cords (mean of two per home). The intervention was associated with a small but significant reduction in four of the five most prevalent hazards. The mean number of unsafe rugs and mats was reduced by 1.57 per house (95% confidence interval (CI) = 0.91 2.24); the mean number of unsafe steps was reduced by 0.61 per house (95% CI = 0.28-0.94); the mean number of rooms with trailing cords was reduced by 0.43 per house (95% CI = 0.10-0.76); and the mean number of unsafe chairs was reduced by 0.10 per house (95% CI = 0.02-0.18). Safety devices were installed in 81.9% of homes. Advice on modifying specific hazards identified on the home hazard assessment resulted in over 50% of subjects removing hazards of floor rugs and mats, trailing cords, and obstacles. The general education message prompted less activity to reduce these hazards than did the advice on identified hazards. CONCLUSIONS: Fall hazards are ubiquitous in the homes of older people. The intervention resulted in a small reduction in the mean number of hazards per house, with many study subjects taking action but removing only a few hazards. The impact of the intervention in achieving self-reported action to reduce hazards was high. PMID- 11890582 TI - Preventing falls in older people: outcome evaluation of a randomized controlled trial. AB - OBJECTIVES: To evaluate the outcome of an intervention to reduce hazards in the home on the rate of falls in seniors. DESIGN: Randomized controlled trial, with follow-up of subjects for 1 year. SETTING: Community-based study in Perth, Western Australia. PARTICIPANTS: People age 70 and older. INTERVENTION: One thousand eight hundred seventy-nine subjects were recruited and randomly allocated by household to the intervention and control groups in the ratio 1:2. Because of early withdrawals, 1,737 subjects commenced the study. All members of both groups received a single home visit from a research nurse. Intervention subjects (n = 570) were offered a home hazard assessment, information on hazard reduction, and the installation of safety devices, whereas control subjects (n = 1,167) received no safety devices or information on home hazard reduction. MEASUREMENTS: Both groups recorded falls on a daily calendar. Reported falls were confirmed by a semistructured telephone interview and were assigned to one of three overlapping categories: all falls, falls inside the home, and falls involving environmental hazards in the home. Analysis was by multivariate modelling of rate ratios and odds ratios for falls, corrected for household clustering, using Poisson regression and logistic regression with robust variance estimation. RESULTS: Overall, 86% of study subjects completed the 1 year of follow-up. The intervention was not associated with any significant reduction in falls or fall-related injuries. There was no significant reduction in the intervention group in the incidence rate of falls involving environmental hazards inside the home (adjusted rate ratio, 1.11; 95% CI = 0.82-1.50), or the proportion of the intervention group who fell because of hazards inside the home (adjusted odds ratio, 0.97; 95% CI = 0.74-1.28). No reduction was seen in the rate of all falls (adjusted rate ratio, 1.02; 95% CI = 0.83-1.27) or the rate of falls inside the home (adjusted rate ratio, 1.17; 95% CI = 0.85-1.60). There was no significant reduction in the rate of injurious falls in intervention subjects (adjusted rate ratio, 0.92; 95% CI = 0.73-1.14). CONCLUSIONS: The intervention failed to achieve a reduction in the occurrence of falls. This was most likely because the intervention strategies had a limited effect on the number of hazards in the homes of intervention subjects. The study provides evidence that a one time intervention program of education, hazard assessment, and home modification to reduce fall hazards in the homes of healthy older people is not an effective strategy for the prevention of falls in seniors. PMID- 11890583 TI - Associations of demographic, functional, and behavioral characteristics with activity-related fear of falling among older adults transitioning to frailty. AB - OBJECTIVES: To determine, in a cohort of older individuals transitioning to frailty (defined by Speechley and Tinetti, 1991) who have previously fallen, whether there are significant associations between demographic, functional, and behavioral characteristics and activity-related fear of falling, using both the Falls Efficacy Scale (FES) and the Activities-Specific Balance Confidence Scale (ABC). DESIGN: Baseline cross-sectional analysis in a prospective cohort intervention study. SETTING: Twenty independent senior living facilities in Atlanta. PARTICIPANTS: Seventeen male and 270 female subjects (n = 287), age 70 and older (mean +/- standard deviation, 80.9 +/- 6.2), with Mini-Mental State Examination score > or = 24, transitioning to frailty, ambulatory (with or without assistive device), medically stable, and having fallen in the past year. MEASUREMENTS: Activity-related fear of falling was evaluated with the FES and ABC Scale. Because of the comparable data derived from each scale, associations with functional measures-related analyses were expressed using the latter. Depression was measured by Center for Epidemiological Studies Depression Scale. Functional measurements included timed 360 degrees turn, functional reach test, timed 10 meter walk test, single limb stands, picking up an object, and three chair stands. RESULTS: No statistically significant association was found between activity-related fear of falling and age. For the proposed activities, about half (ABC, 48.1%; FES, 50.1%) of the subjects were concerned about falling or showed lack of confidence in controlling their balance. A statistically significant inverse correlation was found between FES and ABC (r = -0.65; P < .001). African American subjects showed more activity-related fear of falling than did Caucasians (odds ratio (OR): 2.7 for ABC; 2.1 for FES). Fearful individuals were more likely to be depressed and more likely to report the use of a walking aid than were nonfearful individuals. Fear of falling was significantly correlated to all of the functional measurements (P < .05). In a multivariable logistic regression model, depression, using a walking-aid, slow gait speed, and being an African-American were directly related to being more fearful of falling. CONCLUSIONS: Activity-related fear of falling was present in almost half of this sample of older adults transitioning to frailty. The significant association of activity-related fear of falling with demographic, functional, and behavioral characteristics emphasizes the need for multidimensional intervention strategies to lessen activity-related fear of falling in this population. PMID- 11890584 TI - Progressive versus catastrophic loss of the ability to walk: implications for the prevention of mobility loss. AB - OBJECTIVES: Loss of mobility is an important functional outcome that can have devastating effects on quality of life and the ability of older persons to remain independent in the community. Although a large amount of research has been done on risk factors for disability onset, little work has focused on the pace of disability progression. This study characterizes the development of severe walking disability over time and evaluates risk factors and subsequent mortality as they relate to mobility disability with progressive or catastrophic onset. DESIGN: Population-based prospective cohort study with annual follow-up assessments for up to 7 years SETTING: Three communities of the Established Populations for Epidemiologic Studies of the Elderly. PARTICIPANTS: There were 5,355 persons not disabled at baseline and the first follow-up who had adequate data available to classify mobility disability during subsequent follow-ups. MEASUREMENTS: Severe mobility disability was defined as the need for help from a person to walk across a room or inability to walk across a room. Those developing severe mobility disability were classified as having progressive mobility disability if they had been unable to walk half a mile in either of the prior 2 years. They were classified as having catastrophic mobility disability if they reported having been able to walk half a mile in two previous annual interviews. RESULTS: The overall incidence of severe mobility disability was 11.6 cases/1,000 person years. Those age 85 and older or having three or more chronic conditions at baseline were significantly more likely to develop progressive disability than catastrophic disability. Stroke, hip fracture, and cancer occurring during follow up were associated with very high risk of severe mobility disability. For stroke and hip fracture, the risk was twice as high for catastrophic disability as for progressive disability, but this difference did not reach statistical significance. Risk for catastrophic disability from cancer was significantly greater than for progressive disability. Half of catastrophic disability subjects had stroke, hip fracture, or cancer in the year immediately preceding this disability. Incident heart attack did not predict severe mobility disability. Among those who developed severe mobility disability, type of disability did not influence subsequent survival for the first 3 years, but beyond 3 years those with catastrophic disability had a relative risk of death of 0.4 (95% confidence interval 0.2-0.9) compared with those with progressive disability. CONCLUSION: The observation that risk factors and mortality outcomes were both different for progressive and catastrophic mobility disability supports the value of ascertaining the pace of disability development as a useful characterization of disability. Further progress in developing prevention and treatment strategies may be made by taking the pace of disability development into account. PMID- 11890585 TI - Medical comorbidity and rehabilitation efficiency in geriatric inpatients. AB - OBJECTIVES: To measure and describe medical comorbidity in geriatric rehabilitation patients and investigate its relationship to rehabilitation efficiency. DESIGN: Prospective, multivariate, within-subject design. SETTING: The Geriatric Rehabilitation inpatient unit of the SCO Health Service in Ottawa, Canada. PARTICIPANTS: One hundred ten patients, with a mean age of 82 years. MEASUREMENTS: The rehabilitation efficiency ratio, based on gains in functional status achieved with rehabilitation treatment, and the length of stay were computed for all patients. Values were regressed on the scores of the Cumulative Illness Rating Scale (CIRS), the Mini-Mental State Examination, and the Geriatric Depression Scale to establish predictive power. RESULTS: The findings suggest that geriatric rehabilitation patients experience considerable medical comorbidity. Sixty percent of patients had impairments across six of the 13 dimensions of the CIRS, whereas 36% of patients had impairments across 11 of the 13 dimensions. In addition, medical comorbidity was negatively related to rehabilitation efficiency. This relationship was significant even after controlling for age, cognitive status, depressive symptoms, and functional independence status at admission. CONCLUSION: Medical comorbidity was a significant predictor of rehabilitation efficiency in geriatric patients. Comorbidity scores >5 were prognostic of poorer rehabilitation outcomes and can serve as an empirical guide in estimating a patient's suitability for rehabilitation. Medical comorbidity predicted both the overall functional change achieved with retabilitation (Functional Independence Measure gains) and the rate at with which those gains were reached (rehabilitation efficiency ratio). PMID- 11890586 TI - Effects of aging on hand function. AB - OBJECTIVES: The purpose of this study was to quantify age-induced changes in handgrip and finger-pinch strength, ability to maintain a steady submaximal finger pinch force and pinch posture, speed in relocating small objects with finger grip, and ability to discriminate two identical mechanical stimuli applied to the finger tip. DESIGN: A cross-sectional study. SETTINGS: Greater Cleveland area of Ohio. PARTICIPANTS: Healthy, independent, young (n = 27, range 20-35 years) and older (n = 28, range 65-79 years) subjects. MEASUREMENTS: Handgrip strength, maximum pinch force (MPF), ability to maintain a steady pinch force at three relative force levels (5%, 10%, and 20% MPF) and three absolute force levels (2.5 Newtons (N), 4 N, and 8 N), ability to maintain a precision pinch posture, speed in relocating pegs from a nearby location onto the pegboard, and the shortest distance for discriminating two stimuli were measured in both young and older groups. RESULTS: Compared with young subjects, the older group's handgrip force was 30% weaker (P < .001), MPF was 26% lower (P < .05), and ability to maintain steady submaximal pinch force and a precision pinch posture was significantly less (P < .05). The time taken to relocate the pegs and the distance needed to discriminate two identical stimuli increased significantly with age (P < .01). The decrease in the ability to maintain steady submaximal pinch force was more pronounced in women than men. CONCLUSION: Aging has a degenerative effect on hand function, including declines in hand and finger strength and ability to control submaximal pinch force and maintain a steady precision pinch posture, manual speed, and hand sensation. PMID- 11890587 TI - The association of menopause and physical functioning in women at midlife. AB - OBJECTIVES: To determine whether the characteristics of menstrual bleeding and the menopausal transition are associated with physical functioning in women age 40 to 55, after considering ethnicity, ability to pay for basics, body size, and age. DESIGN: Cross-sectional study. SETTING: Seven geographically dispersed community samples in the United States. PARTICIPANTS: The 14,427 respondents were Caucasians (46.9%), African Americans (28.7%), Chinese (4.0%), Japanese (5.3%), and Hispanics (12.6%) from the Study of Women's Health Across the Nation (SWAN) Cross-sectional Study, a study of the menopausal transition, including surgical menopause. MEASUREMENTS: The dependent variable was a three-category variable based on the physical functioning scale of the Medical Outcomes Study. Explanatory variables included menstrual and menopausal status. RESULTS: Eighty percent (80.8%) of women reported no limitation in physical functioning, whereas 10% of women had some limitation, and 9.2% of women indicated having substantial limitation. Women with substantial limitation in physical functioning had double the prevalence odds ratio (POR = 2.02; 95% confidence interval (CI) = 1.64-2.49) of having surgical menopause and 76% greater odds (POR = 1.76; 95% CI = 1.38 2.24)) of using hormones, compared with women with no limitation. Compared with those without limitation, women with substantial limitation in physical functioning had 56% greater odds (POR = 1.56; 95% CI = 1.23-1.97)) of being naturally postmenopausal and a 41% greater odds (POR = 1.41; 95% CI = 1.17-1.70) of being perimenopausal, relative to being premenopausal and after adjusting for other variables. CONCLUSION: Even at the relatively early age of 40 to 55, approximately 20% of women self-reported limitation in physical functioning. Surgical menopause and the use of hormones were more frequently observed in women with some and substantial physical limitation than in women with no limitation, even after adjusting for economic status, age, body mass index, and race/ethnicity. PMID- 11890588 TI - Does hospice have a role in nursing home care at the end of life? AB - OBJECTIVES: To assess the possible benefits and challenges of hospice involvement in nursing home care by comparing the survival and needs for palliative care of hospice patients in long-term care facilities with those living in the community. DESIGN: Retrospective review of computerized clinical care records. SETTING: A metropolitan nonprofit hospice. PARTICIPANTS: The records of 1,692 patients were searched, and 1,142 patients age 65 and older were identified. Of these, 167 lived in nursing homes and 975 lived in the community. MEASUREMENTS: Patient characteristics, needs for palliative care, and survival. RESULTS: At the time of enrollment, nursing home residents were more likely to have a Do Not Resuscitate order (90% vs 73%; P < .001) and a durable power of attorney for health care (22% vs 10%; P < .001) than were those living in the community. Nursing home residents also had different admitting diagnoses, most notably a lower prevalence of cancer (44% vs 74%; P < .032). Several needs for palliative care were less common among nursing home residents, including constipation (1% vs 5%; P = .02), pain (25% vs 41%; P < .001), and anticipatory grief (1% vs 9%; P < .001). Overall, nursing home residents had fewer needs for care (median 0, range 0-3 vs median 1, range 0 5; rank sum test P < .001). Nursing home residents had a significantly shorter survival (median 11 vs 19 days; log rank test of survivor functions P < .001) and were less likely to withdraw from hospice voluntarily (8% vs 14%; P = .03). However, there was no difference in the likelihood of becoming ineligible during hospice enrollment (6% for both groups). CONCLUSIONS: These results suggest that hospices identify needs for palliative care in a substantial proportion of nursing home residents who are referred to hospice, although nursing home residents may have fewer identifiable needs for care than do community-dwelling older people. However, the finding that nursing home residents' survival is shorter may be of concern to hospices that are considering partnerships with nursing homes. An increased emphasis on hospice care in nursing homes should be accompanied by targeted educational efforts to encourage early referral. PMID- 11890589 TI - Advancing age and cervical cancer screening and prognosis. AB - OBJECTIVES: To determine associations between advancing age and screening behavior and prognosis in long-term members of a prepaid health plan diagnosed with invasive cervical cancer (ICC). DESIGN: Case series. SETTING: Prepaid health plan. PARTICIPANTS: All women diagnosed with ICC at Kaiser Permanente Medical Care Program-Northern California health plan from 1988 to 1994. MEASUREMENTS: From medical records, we recorded participants' age, stage at diagnosis, tumor histology, and results of and reasons for all previous cervical smears. We limited our analysis to women who had been members of the health plan for at least 30 of the 36 months preceding diagnosis (n = 455). RESULTS: Women in older age groups were less likely than younger women to have been screened within the 3 years before diagnosis (P = .005 for trend). Nonadherence to follow-up of abnormal cervical smears was uncommon (17/455, 3.7%) and not age related (P = .932 for trend). The proportions of ICC that were interval cancers, defined as ICC diagnosed within 3 years of a negative screening smear, were highest in women under age 30 (P = .004 for trend). In multivariate analyses controlling for stage at diagnosis, women age 60 and older were not more likely to die of ICC within 3 years of diagnosis than were women younger than age 60 (odds ratio 1.30, 95% confidence interval 0.75-2.28). CONCLUSION: The disproportionate burden of cervical cancer observed in older women appears to be largely attributable to lack of screening within the 3 years before diagnosis. PMID- 11890590 TI - The effects of patient communication skills training on the discourse of older patients during a primary care interview. AB - OBJECTIVES: To test the effects of a communication skills training intervention on older patients' discourse during a primary care interview. DESIGN: A quasi experimental design involving two intervention conditions. SETTING: The Family Practice Center of a university-based clinic. PARTICIPANTS: Thirty-three patients averaging age 72 and 9 family practice physicians. INTERVENTION: A communication skills training booklet received approximately 3 days before the scheduled appointment and a 30-minute face-to-face follow-up session before seeing the physician. MEASUREMENTS: Patients' seeking, providing, and verifying of information were coded from transcripts of the 33 interviews. RESULTS: Trained patients engaged in significantly more seeking and providing of information than untrained patients. Additionally, trained patients obtained significantly more information from physicians than did untrained patients, both in terms of the number of total information units and the number of units per question asked. CONCLUSION: Patient communication skills training appears to be an effective means of enhancing patients' participation in the medical interview without increasing the overall length of the interview. PMID- 11890591 TI - Genetic and environmental influences on self-reported reduced hearing in the old and oldest old. AB - OBJECTIVES: The aim of the present twin study was to estimate the relative importance of genetic and environmental factors in variation in self-reported reduced hearing among the old and the oldest old. DESIGN: Self-reported hearing abilities of older twins assessed at intake interview in a population-based longitudinal survey. SETTING: Denmark. PARTICIPANTS: Twins age 75 and older identified in the population-based Danish Twin Registry in 1995. An interview was conducted with 77% of 3,099 individuals in the study population. In 1997 and 1999, a follow-up contact to the survivors was made and an additional 2,778 twins, age 70-76, were included in the study. MEASUREMENTS: Reduced hearing was assessed by the same question in all interview waves. Heritability (proportion of the population variance attributable to genetic variation) was estimated using structural-equation analyses. RESULTS: The prevalence of self-reported reduced hearing corresponded to previous studies and showed the expected age and sex dependence. Concordance rates, odds ratios, and correlations were consistently higher for monozygotic twin pairs than for dizygotic twin pairs in all age and sex categories, indicating heritable effects. Structural-equation analyses revealed a substantial heritability for self-reported reduced hearing of 40% (95% CI = 19-53%). The remaining variation could be attributed to individuals' nonfamilial environments. CONCLUSION: We found that genetic factors play an important role in self-reported reduced hearing in both men and women age 70 and older. Because self-reports of reduced hearing involve misclassification, this estimate of the genetic influence on hearing disabilities is probably conservative. Hence, genetic and environmental factors play a substantial role in reduced hearing among the old and oldest old. This suggests that clinical epidemiological studies of age-related hearing loss should include not only information on environmental exposures but also on family history of hearing loss and, if possible, biological samples for future studies of candidate genes for hearing loss. PMID- 11890592 TI - Effects of aging on the opioid modulation of feeding in humans. AB - OBJECTIVES: To determine whether aging is associated with a reduction in the opioid modulation of feeding, which may be important in the pathogenesis of the "anorexia of aging." DESIGN: Three studies on separate days, in randomized order and double-blind fashion. SETTING: Clinical Human Research Laboratory, Department of Medicine, RAH, Adelaide, Australia. PARTICIPANTS: Twelve older (5 male/7 female) (age 65-84) and 12 young (5 male/7 female) (age 20-26) healthy subjects. INTERVENTION: Subjects received in double-blinded random order, intravenous bolus (10 minutes) and then continuous (140 minutes) infusions of saline (control), naloxone low dose (LD) (bolus 27 microg/kg; continuous 50 microg/kg/hr), or naloxone high dose (HD) (bolus 54.5 microg/kg; continuous 100 microg/kg/hr). MEASUREMENTS: After 120 minutes, subjects were offered a buffet meal, and their energy intake was quantified. Hunger, fullness, nausea, and drowsiness were assessed using visual analogue scales. RESULTS: The naloxone LD and HD infusions had no significant effect on ratings of hunger, fullness, or nausea, but increased drowsiness (P < .01) compared with the control infusion in both age groups. Older subjects ate less (P < .001) at the buffet meal than young subjects during all three infusions. Naloxone infusions reduced energy intake compared with control (P < .001), LD by 13.2 +/- 5.0% and HD by 10.7 +/- 5.0%, with no difference between the doses (P = .71). Overall, naloxone suppressed energy intake in both young and older subjects (P < .01). This suppression was slightly, but not significantly, greater in young than in older subjects (mean of LD and HD 16.4 +/- 4.9% vs 7.5 +/- 4.9%, P = .42), because of a trend to reduced suppression in older women. CONCLUSIONS: We conclude that healthy older adults retain their sensitivity to the suppressive effects of naloxone on food intake. Possible gender differences in this sensitivity warrant further investigation. A decline in opioid activity is unlikely to contribute substantially to the physiological anorexia of aging observed in older people. PMID- 11890593 TI - Healthcare costs associated with percutaneous endoscopic gastrostomy among older adults in a defined community. AB - OBJECTIVES: The effectiveness of percutaneous endoscopic gastrostomy (PEG) in older adults remains controversial. Although prior studies have examined the safety of PEG and its impact on nutrition, there are limited data on the economic costs. The purpose of this study is to describe the healthcare costs associated with PEG tube feeding over 1 year. DESIGN: Prospective cohort study. SETTING: Small community of approximately 60,000 residents served by two hospital systems. PARTICIPANTS: One hundred five (70%) of 150 patients age 60 and older receiving PEG over a 24-month period in the targeted community who permitted access to their medical records. MEASUREMENTS: Patients were interviewed at baseline and every 2 months for 1 year to obtain information on use of enteral formula, complication rates, and health services use. We obtained inpatient charge data for all hospitalizations and PEG procedures for 1 year. RESULTS: Censoring patients at death or 1 year post-PEG, the mean number of days of PEG tube feeding was 180 (range 5-365). The average cost for PEG tube feeding for this cohort of patients was $7,488 (median $3,691) in 1997 and 1998. The average daily cost of PEG tube feeding was $87.21 (median $33.50). The estimated cost of providing 1 year of feeding via PEG is $31,832 (median $12,227). The main components of this cost include the initial PEG procedure (29.4%), enteral formula (24.9%), and hospital charges for major complications (33.4%). CONCLUSIONS: Direct charges associated with PEG tube feeding over 1 year are conservatively estimated at $31,832; there was considerable variation in charges because of the cost of rare but costly major complications. Also, feeding patients via PEG resulted in cost shifts in terms of the primary payor. The economic cost of PEG tube feeding is another consideration in decision making for long-term enteral feeding among older adults. PMID- 11890594 TI - How EverCare nurse practitioners spend their time. AB - OBJECTIVES: To describe how nurse practitioners (NPs) employed by EverCare, a Medicare HMO serving exclusively nursing home residents, spend their working days. DESIGN: A descriptive study based on structured self-reports. SETTING: Nursing homes. PARTICIPANTS: Seventeen NPs employed by EverCare in five sites. MEASUREMENTS: Self-reports of time spent over a 2-week period and specific reports of how time was spent on selected cases. RESULTS: NPs spend about 35% of their working day on direct patient care and another 26% in indirect care activities. Of the latter, 46% of the time was spent interacting with nursing home staff, 26% with family, and 15% with the physicians. The mean time spent on a given patient per day was 42 minutes (median 30); of this time 20 minutes was direct care (median 15). CONCLUSIONS: NPs' activities are varied. Much of their time is spent communicating with vital parties, an important function that supports the physicians' primary care role and should enhance families' satisfaction with care. PMID- 11890595 TI - Geriatricians health survey 2000. AB - OBJECTIVES: To characterize geriatricians' preventive health behaviors including vitamin/supplement use, exercise, smoking, alcohol use, and weight control. DESIGN: Mailed questionnaire. SETTING: United States. PARTICIPANTS: Two thousand six hundred eleven U.S. physicians certified as having added qualifications in geriatric medicine and who were members in the American Geriatrics Society; 1,524 returned completed questionnaires (58%). MEASUREMENTS: Rates of supplement use and recommendations, preventive health visits, advance directive completion, exercise, religious service attendance, smoking, alcohol use, and amount of adult weight gain. RESULTS: Most responding geriatricians took at least one vitamin supplement: 50% vitamin E, 50% a multivitamin (MVI), and 31% vitamin C. Calcium ingestion was common among women. Other supplement use was uncommon: ginkgo compounds were consumed by 47 (3%), and 77 (5%) took a variety of other nonvitamin supplements. Over 90% recommended vitamins, especially multivitamins and vitamin E, at least sometimes. Recommendations for ginkgo (38%) and St. John's wort (33%) were also common. Almost half of respondents had completed a formal advance directive. Exercise was practiced at least weekly by 88%. Cigarette smoking was rare (1%), but at least occasional alcohol use was common (85%). Most of respondents were men (74%), and 35% had completed fellowship training. CONCLUSION: Vitamin/supplement use was common among responding geriatricians but not universal. Respondents often recommended MVI, vitamin E, and vitamin C, but were less likely to consume or recommend other supplements. The most common preventive health behavior among our respondents was exercise. PMID- 11890597 TI - Measuring fitness in healthy older adults: the Health ABC Long Distance Corridor Walk. AB - OBJECTIVES: The Health ABC Long Distance Corridor Walk (LDCW) was designed to extend the testing range of self-paced walking tests of fitness for older adults by including a warm-up and timing performance over 400 meters. This study compares performance on the LDCW and 6-minute walk to determine whether the LDCW encourages greater participant effort. DESIGN: Subjects were administered the LDCW and 6-minute walk during a single visit. Test order alternated between subjects, and a 15-minute rest was given between tests. SETTING: The Baltimore Veterans Affairs Medical Center. PARTICIPANTS: Twenty volunteers age 70 to 78. MEASUREMENTS: The LDCW, consisting of a 2-minute warm-up walk followed by a 400 meter walk and a 6-minute walk test were administered using a 20-meter long course in an unobstructed hallway. Heart rate (HR) and blood pressure (BP) were recorded at rest and before and after all walks. RESULTS: All 20 subjects walked a faster pace over 400 meters than for 6 minutes, in which the mean distance covered was 402 meters. From paired t-tests, walking speed was faster (mean difference = 0.23 m/sec; P < .001), and ending HR (mean difference = 7.6 bpm; P < .001) and systolic BP (mean difference = 8.3 mmHg; P = .024) were greater for the 400-meter walk than for the 6-minute walk. Results were independent of test order and subject fitness level. CONCLUSIONS: Providing a warm-up walk and using a target distance instead of time encouraged subjects to work closer to their maximum capacity. This low-cost alternative to treadmill testing can be used in research and clinical settings to assess fitness and help identify early functional decline in older adults. PMID- 11890596 TI - Persistent mobility deficit in the absence of deficits in activities of daily living: a risk factor for mortality. AB - OBJECTIVES: To investigate the extent to which self-reported mobility deficit in the absence of impairment in activities of daily living (ADL) is associated with elevated mortality risk. DESIGN: Prospective cohort study, with annual assessments of mobility and ADL status and ongoing monitoring of vital status. SETTING: Population-based cohort drawn from Medicare enrollees in New York City. PARTICIPANTS: One thousand two hundred ninety-eight older adults reporting functional status at baseline (1992-1994) and 2 years later. MEASUREMENTS: Subjects reported mobility (e.g., walking, climbing stairs, and rising from a chair) and ADL (e.g., bathing, toilet use, dressing, grooming, and feeding) limitations. Two-year functional status trajectories were noted. We used two additional follow-up periods, at 2 and 4 years, to examine the likelihood that older people with mobility deficit may face an increased risk of death without first passing through a state of enduring ADL disability. RESULTS: At 2 years, 12.7% had incident mobility deficit without ADL disability, and 21.3% were persistently disabled in mobility without ADL disability. Relative to subjects free of disability at baseline and follow-up, risk of mortality in the incident mobility deficit group was elevated at 2 and 4 years but did not achieve statistical significance. By contrast, for subjects with persistent mobility impairment who did not report ADL impairment, the mortality risk was significantly elevated both at 2 years (relative risk (RR) = 2.5; 95% confidence interval (CI) = 1.1-5.7)) and 4 years (RR = 2.9; 95% CI = 1.7-4.9)) of follow-up. Mortality was significantly elevated in this group in analyses restricted to respondents with no or only one comorbid condition. CONCLUSION: Continuing, self reported mobility impairment in the absence of ADL deficit is a risk factor for mortality. Older people with self-reported mobility deficit face an increased risk of mortality without first passing through enduring states of ADL disability. PMID- 11890598 TI - Elective discontinuation of life-sustaining mechanical ventilation on a chronic ventilator unit. AB - Withdrawal of medical interventions has become common in the hospital for patients with terminal disease. Despite the widespread feeling that medical interventions may be futile in certain patients, many patients, families, and medical staff find withdrawal of care difficult and withdrawal of mechanical ventilation to be the most disturbing secondary to the close proximity of withdrawal and death. Presented is a 6-year retrospective review of elective withdrawal of life-sustaining mechanical ventilation on a chronic ventilator unit (CVU) in an academic nursing home. Of the 98 patients admitted to the 19-bed CVU during this period, only 13 underwent terminal weaning (TW). Statistically, these 13 patients did not differ significantly in age, gender, race, route of nutrition, decisional capacity, or length of stay on the unit compared with the 85 patients who were not terminally weaned (t-test P > .05). Stepwise logistic regression found that patients who were more alert at admission were more likely to have participated in TW (chi2 = 5.22, coefficient for alertness P < .036). The decision to terminate mechanical ventilation was made by patients in eight cases and by family in five cases. The first step in the process leading to TW was a discussion with the patient and family about plan of care, including the patient's desires for attempted resuscitation, rehospitalization, advance directives, and family contacts. Plan of care was reviewed informally in a weekly multidisciplinary round and formally, to address each patient's care plan, in a multidisciplinary family meeting on a regular basis. The second step was to address TW when brought up by the patient, family, or medical staff. A request for TW by a patient or surrogate was referred to the medical staff, who screened the patient for depression or other remediable symptoms. The third step was to refer the patient and family to another formal meeting to discuss the request for TW and, if needed, in the case of multiple family members, to allow questions to be answered and consensus to be formed. Additional meetings were scheduled as needed. The next step occurred once a consensus was reached to proceed with TW; a date and time was set to reconvene the patient, family, and anyone else who wanted to be present at the TW. The TW process began when a peripheral intravenous catheter was placed and the patient was premedicated with low doses of morphine sulfate and a benzodiazepine. After premedication, the patient was removed from the ventilator. The physician, nurse, family, and physician assistant remained at the bedside and additional morphine or benzodiazepine was given, as needed, for symptom management. Death from TW occurred in all patients, at times ranging from 2 minutes to 10.5 hours (average 6.2 hours). A mean total dose of 115 mg morphine and 14 mg diazepam was given for symptom control. There was no correlation between dose of these medications and duration of survival off the ventilator. PMID- 11890599 TI - Popular views of old age in America, 1900-1950. AB - The aging of the American population has significantly changed medical practice over the last century. As is well known, life expectancy first began to increase dramatically in the late 19th century, but at the same time that the numbers of older people have been increasing, the social and cultural meanings of growing old have also changed. It is likely that different cohorts of older people have had different experiences with old age because of the time periods they lived through. This paper offers one way to look at some of the historical changes that have affected the public and the medical profession on the subject of old age by looking at old age through American popular literature in the first half of the 20th century in three overlapping time periods. In the first three decades of the century, the concept of old age was widely defined, and older people (rather than physicians) were the principal authorities in describing the qualities of old age. In the third and fourth decades of the century, the idea of old age was starting to acquire increasing negative connotations, but chronological age itself did not signal the beginning of old age. However, by the late 1930s and 1940s, old age became widely viewed as a specific social and medical problem to be addressed by professionals, and older people had become a recognizable population, with a variety of groups organized around their care. This paper illustrates changes in American understandings of old age within and without the medical profession and suggests ways in which popular conceptions of old age might continue to shift and affect how physicians take care of their older patients in the future. PMID- 11890600 TI - Outcomes in older people undergoing operative intervention for colorectal cancer. PMID- 11890601 TI - Osteoprotective benefits of exercise: more pain, less gain? AB - Based on the results of their study, Hagberg et al. suggested that physical activity that is moderate in intensity might be most appropriate for the prevention of osteoporosis in postmenopausal women. The take-home message is that being physically active has osteoprotective benefits. However, the implication that more vigorous exercise is less beneficial must be carefully scrutinized. When compared with the physically active nonathletes, the highly trained athletes in this study tended to be more years beyond menopause, had been on HRT for a smaller proportion of the postmenopausal period, and may have had lower endogenous estrogen levels as a result of having less fat mass. The chronic exposure of the skeleton to estrogens very likely played a more prominent role in determining BMD status in the women in this study than the investigators were able to ascertain, owing largely to the cross-sectional nature of the investigation. It is safe to conclude that an exercise prescription for the prevention of osteoporosis should include weight-bearing exercise, but the optimal intensity, frequency, and duration of exercise remain to be established. The results of the study by Hagberg et al. emphasize the importance of physical activity for sustaining BMD, but should not provide a basis for discouraging women from performing vigorous exercise. PMID- 11890602 TI - What heterogeneity among centenarians can teach us about genetics, aging, and longevity. PMID- 11890604 TI - Regarding weight outcomes in antidepressant users in nursing facilities. PMID- 11890603 TI - Palliative care in long-term-care facilities--a comprehensive model. PMID- 11890605 TI - Undertreatment of cardiovascular disease in ethnically diverse older adults: who should receive an electrocardiogram? PMID- 11890606 TI - Blood pressure reduction after the first dose of captopril and perindopril. PMID- 11890607 TI - Risk factors for developing cardiac disease in late middle-aged and older men and women: a prospective study. PMID- 11890608 TI - Alternative medicine use in older Americans. PMID- 11890609 TI - Atypical presentation of silent nocturnal hypoglycemia in an older person. PMID- 11890610 TI - The effect of low-dose daily aspirin intake on survival in the Finnish centenarians cohort. PMID- 11890611 TI - Antigen-induced T cell death is regulated by CD4 expression. AB - Activation induced cell death (AICD) is a major physiologic pathway that regulates T cell homeostasis. In CD4 T cells, AICD is mediated mainly through Fas/FasL interactions. Although TCR occupancy triggers AICD, the contribution of its tightly associated CD4 coreceptor to the process that leads to AICD is not known. Here we show that CD4 molecule plays an essential regulatory role of TCR dependent AICD. Loss of CD4 rendered activated 5kc T cell hybridoma resistant to AICD. The resistance of CD4 negative 5kc T cells to AICD was due to selective inhibition of FasL expression and it could be reversed by addition of recombinant FasL. Furthermore, a direct functional link between CD4 and FasL was demonstrated by induction of FasL upon CD4 crosslinking in a TCR independent fashion. The importance of CD4 interaction with MHC/peptide complex in mediating AICD was also evident in normal T cells that could survive chronic stimulation with anti-CD3 but died after short period of proliferation after stimulation with MHC/peptide. Thus it appears that AICD is controlled by the CD4 molecule via regulation of FasL expression. These findings have important implications for our understanding of mechanisms of peripheral tolerance as well as pathogenesis of autoimmune diseases. PMID- 11890613 TI - Self peptides and the peptidic self. AB - Twenty years ago, antigenic and self peptides presented by MHC molecules were absent from the immunological scene. While foreign peptides could be assayed by immune reactions, self peptides, as elusive and invisible as they were at the time, were bound to have an immunological role. How self peptides are selected and presented by MHC molecules, and how self MHC-peptide complexes are seen or not seen by T cells raised multiple questions particularly related to MHC restriction, alloreactivity, positive and negative selection, the nature of tumor antigens and tolerance. These issues were addressed in the "peptiditic self model" (1986) and subsequent hypothesis. They are retrospectively and critically reviewed here in the context of our current understanding of these major immunological phenomena. PMID- 11890612 TI - Modulation of CD4 T cell function by soluble MHC II-peptide chimeras. AB - Peptides antigens of 8 to 24 amino acid residues in length that are derived from processing of foreign proteins by antigen presenting cells (APC), and then presented to T cells in the context of major histocompatibility complex molecules (MHC) expressed by APC, are the only physiological ligands for T cell receptor (TCR). Co-ligation of TCR and CD4 co-receptor on T cells by MHC II-peptide complexes (signal 1) leads to various T cell functions depending on the nature of TCR and CD4 co-ligation, and whether costimulatory receptors (signal 2) such as CD28, CTLA-4, CD40L are involved in this interaction. Recently, the advance of genetic engineering led to the generation of a new class of antigen-specific ligands for TCR, i.e., soluble MHC class I-, and MHC class II-peptide chimeras. In principle, these chimeric molecules consist of an antigenic peptide which is covalently linked to the amino terminus of alpha-chain in the case of MHC I, or beta-chains in the case of MHC II molecules. Conceptually, such TCR/CD4 ligands shall provide the signal 1 to T cells. Since soluble MHC-peptide chimeras showed remarkable regulatory effects on peptide-specific T cells in vitro and in vivo, they may represent a new generation of immunospecific T cell modulators with potential therapeutic applicability in autoimmune and infectious diseases. This review is focused on the immunomodulatory effects of soluble, MHC class II peptide chimeras, and discuss these effects in the context of the most accepted theories on T cell regulation. PMID- 11890615 TI - Studies on CD4 T cell immunity using somatic transgene immunization. AB - This review presents and discusses our recent data on the use of somatic transgene immunization in the induction of CD4 T cell responses in vivo. Somatic transgene immunization is a process that targets B lymphocytes resident in the spleen with DNA coding for antigenized immunoglobulin H chain genes. After transgenesis B lymphocytes function as antigen-producing and antigen-presenting cells. The studies reviewed herein describe the characteristics of the primary and memory CD4 response against a dominant Th cell determinant. In addition, they show how ad hoc modifications of the transgene result in the induction of a CD4 T cell response against Th cell determinants against which a response is normally not obtained. The new concept Th-Th cooperation is discussed. PMID- 11890614 TI - Multi-modal antigen specific therapy for autoimmunity. AB - Peripheral tolerance, represents an attractive strategy to down-regulate previously activated T cells and suppress an ongoing disease. Herein, immunoglobulins (Igs) were used to deliver self and altered self peptides for efficient peptide presentation without costimulation to test for modulation of experimental allergic encephalomyelitis (EAE). Accordingly, the encephalitogenic proteolipid protein (PLP) sequence 139-151 (referred to as PLP1) and an altered form of PLP1 known as PLP-LR were genetically expressed on Igs and the resulting Ig-PLP1 and Ig-PLP-LR were tested for efficient presentation of the peptides and for amelioration of ongoing EAE. Evidence is presented indicating that Ig-PLP1 as well as Ig-PLP-LR given in saline to mice with ongoing clinical EAE suppresses subsequent relapses. However, aggregation of both chimeras allows crosslinking of Fcgamma receptors (FcgammaRs) and induction of IL-10 production by APCs but does not promote the up-regulation of costimulatory molecules. Consequently, IL-10 displays bystander suppression and synergizes with presentation without costimulation to drive effective modulation of EAE. As Ig-PLP1 is more potent than Ig-PLP-LR in the down-regulation of T cells, we conclude that peptide affinity plays a critical role in this multi-modal approach of T cell modulation. PMID- 11890617 TI - "Troy-bodies": recombinant antibodies that target T cell epitopes to antigen presenting cells. AB - Targeting of antigens to antigen presenting cells (APC) results in enhanced antigen presentation and T cell activation. In this paper, we describe a novel targeting reagent denoted "Troy-bodies", namely recombinant antibodies with APC specific V regions and C regions with integrated T cell epitopes. We have made such antibodies with V regions specific for either IgD or MHC class II, and four different T cell epitopes have been tested. All four epitopes could be introduced into loops of C domains without disrupting Ig folding, and they could be released and presented by APC. Furthermore, whether IgD- or MHC-specific, the molecules enhanced T cell stimulation compared to non-specific control antibodies in vitro as well as in vivo. Using this technology, specific reagents can be designed that target selected antigenic peptides to an APC of choice. Troy-bodies may therefore be useful for manipulation of immune responses, and in particular for vaccination purposes. PMID- 11890616 TI - Gene therapeutic approaches to induction and maintenance of tolerance. AB - Tolerance induction would be an ideal way to treat autoimmune diseases, especially if achievable in primed individuals. Moreover, specific tolerance would preclude the need for immunosuppressive treatment with its side effects. In this review, we will revisit the historical concepts of tolerance, and introduce a novel approach to tolerance via gene therapy. Our model system is based on the tolerogenicity of IgG carriers and B-cell antigen presentation. Results with this model demonstrate that it is simple and effective even in primed recipients, and has proven efficacy in three autoimmune models. We discuss the mechanisms of tolerance with fusion IgG's and analyze how our model of gene therapy approached can be utilized to fit in the future treatment of autoimmune conditions. PMID- 11890618 TI - Changing frequency of parotid gland neoplasms--analysis of 560 tumours treated in a district general hospital. AB - An analysis of all parotidectomies performed for neoplastic lesions in the maxillofacial unit at a district general hospital during a 26-year period between 1974-1999 was undertaken. The details analysed were age, sex, histology and temporal variations in the frequency of specific tumour types during the study period. A total of 538 parotidectomies performed on 529 patients in whom 560 tumours were present, formed the basis of this study. Marked variations were present in the age and sex distribution and relative frequency of specific tumour types in this study, when compared to previous reports. There were also differences in the age and sex distribution of pleomorphic adenoma and adenolymphoma (P <0.0001) in this study. The relative frequency of benign tumours and adenolymphoma increased, whereas that of pleomorphic adenoma decreased during the study period. In addition, there was a statistically significant decrease in the relative frequency of pleomorphic adenoma (P <0.0001) and an increase in adenolymphoma (P <0.0001) when comparisons were made with previous studies. This study from a defined population may be more representative of the true proportion of specific tumours in this population. The potential implications of the results on the investigation and treatment of parotid neoplasms is highlighted. PMID- 11890619 TI - Acute cholecystitis--room for improvement? AB - AIMS: A recent survey of UK general surgeons showed that almost 90% prefer to manage patients with acute cholecystitis by initial conservative management and delayed cholecystectomy (DC). The aim of this study was to assess the effectiveness of this management policy in a large university hospital. PATIENTS AND METHODS: All patients admitted with acute cholecystitis between January 1997 and June 1999 who went on to have a cholecystectomy were identified. Patients were required to have right upper quadrant pain for > 12 h, a raised white cell count and findings consistent with acute cholecystitis on ultrasound to be included in the study. RESULTS: 109 patients were admitted with acute cholecystitis (76 female, 33 male) with a median age of 62 years (range, 22-88 years). Conservative management failed in 16 patients (14.7%) who underwent emergency cholecystectomy due to continuing symptoms (9), empyema (4) and peritonitis (3). Symptoms settled in 93 patients and delayed cholecystectomy was performed without further problems in 66 (60.6%). 27 patients were re-admitted with further symptoms before their elective surgery and, of these, 3 were admitted for a third time before surgical intervention. Ten of the 30 re admission episodes (33%) occurred within 3 weeks of discharge but 15 (56%) occurred more than 2 months after discharge. Elective surgery was undertaken at a median of 10 weeks post-discharge with 67% of operations occurring within 3 months. Mean total hospital stay (days) +/- SEM, for the three groups were: emergency surgery group, 10.21 +/- 0.85; uncomplicated DC group, 12.48 +/- 0.37; re-admitted group, 14.75 +/- 0.71. CONCLUSIONS: The policy of conservative management and DC was successful in 60.6% of cases but 14.7% of patients required emergency surgery and 24.8% were re-admitted prior to elective surgery with a resultant increase in total hospital stay. Performing elective surgery within 2 months of discharge in all cases would have reduced the re-admission rate by 56% and this along with the increased use of early cholecystectomy during the first admission are areas where the treatment of acute cholecystitis could be significantly improved. PMID- 11890620 TI - Transduodenal sphincteroplasty and transampullary septectomy for sphincter of Oddi dysfunction. AB - BACKGROUND: The diagnosis and management of sphincter of Oddi dysfunction are controversial issues. Both surgical and endoscopic series report modest success in the treatment of this condition. There is evidence from endoscopic series that the Milwaukee classification could predict the clinical outcome after sphincterotomy. We reviewed our long-term results of surgical sphincter ablation for sphincter of Oddi dysfunction, in order to correlate outcome with underlining pathology (biliary versus pancreatic) and Milwaukee biliary group classification. PATIENTS AND METHODS: During a 10 year period (1987-1996), 36 patients with either biliary (n = 26) or pancreatic (n = 10) presentation of suspected sphincter of Oddi dysfunction were selected for surgery according to a standard protocol of investigation and management. All patients were classified according to the Milwaukee classification for the biliary group or its version for the pancreatic group and had transduodenal sphincteroplasty and transampullary septectomy. RESULTS: Despite a trend towards a better outcome in the biliary group (good result 62%, moderate 23%, poor 15%) compared to the pancreatic (good result 40%, moderate 40%, poor 20%) the difference was not statistically significant (P = 0.48). Milwaukee classification for the biliary group correlated well with a favourable outcome (P < 0.05). CONCLUSIONS: The modest outcome despite careful patient selection for surgery emphasises the need for more objective diagnostic tools. Milwaukee classification appears to be of good predictive value, and a good result can be anticipated in type I or even type II patients. The trend towards a better outcome in the biliary group may reflect the weakness of a drainage procedure to treat patients with parenchymal pancreatic disease. PMID- 11890621 TI - Cyclic neutropenia and pyomyositis: a rare cause of overwhelming sepsis. AB - Primary pyomyositis is a pyogenic infection of skeletal muscle with abscess formation, which traditionally lacks an identifiable cause. We present a case of pyomyositis for which a cause was established. This was largely due to the fact that the patient was young and fit, enabling him to survive such overwhelming sepsis long enough for cycling of his neutrophil count to become apparent. Having had multiple abscesses drained, he was successfully treated with granulocyte colony stimulating factor and has remained well since. PMID- 11890622 TI - Laser depilation of the natal cleft--an aid to healing the pilonidal sinus. AB - BACKGROUND: Pilonidal disease is common. Excessive hair growth in the natal cleft is thought to be a factor in initiating these sinuses. It is chronic and intermittent in nature and treatment can be difficult. Hair removal by shaving or use of creams is often advised as a compliment to surgical treatments. However, access to the natal cleft can be difficult. Laser removal of hair in the natal cleft is considered as an aid to healing the pilonidal sinus. PATIENTS AND METHODS: Over a 5-year period, 14 patients with recurrent pilonidal disease were treated in our unit with laser depilation. They were all contacted by postal questionnaire, and those with ongoing disease were asked to return to the clinic for evaluation and possible further treatment. RESULTS: All patients returned the postal questionnaire. Of the 14 patients, 4 had on-going disease and received further depilation with the Alexandrite laser. All are now healed with no reported complications. All patients found the procedure painful and received local anaesthetic. CONCLUSIONS: Laser depilation in the natal cleft is by no means a cure for pilonidal disease. Removal of hair by this method represents an alternative and effective method of hair removal and, although long lasting, is only temporary. However, it allows the sinuses to heal rapidly. It is relatively safe, and simple to teach, with few complications. It should thus be considered as an aid to healing the problem pilonidal sinus. PMID- 11890623 TI - 'Leaving a mark'. AB - Biopsies by various means do have a risk of seeding tumour cells into the biopsy track. Even with fine needle and trocar techniques this has been proven to occur. By excising the biopsy track at the time of surgery this risk can be reduced. With needle and trocar techniques there might be little evidence of the puncture when the time of surgery takes place. We advocate using Indian ink to mark the biopsy site. PMID- 11890624 TI - The outcome of drug smuggling by 'body packers'--the British experience. AB - Body packing or internal concealment used by drug dealers to smuggle illicit substances, puts the body packer at risk of both imprisonment and death. We report our experience over a 4 year period from January 1996 to December 1999 of suspects presenting to our hospital (the largest series in Europe). A total of 572 cases were assessed radiographically and 180 were shown to be carrying foreign bodies. The commonest reasons for admission were suspected overdose or gastrointestinal obstruction. Thirty-six cases were admitted, of whom 7 required surgical intervention. No deaths occurred. Of all people detained for smuggling by internal concealment into Britain during this period, 27% were seen in our hospital. These cases may present alone or escorted by Her Majesty's Customs and Excise personnel, and one must be aware of this possibility even when situated away from a port of entry. PMID- 11890625 TI - A comparison between forefoot plaster and wooden soled shoes following Mitchell's osteotomy for hallux valgus. AB - Between 1 October 1997 and 1 November 1998, 43 patients (59 feet) were treated with a standard Mitchell's osteotomy for hallux valgus. Of these, 26 patients (36 feet) were treated postoperatively in a forefoot plaster. The other 17 patients (23 feet) were treated with a wooden soled shoe. There was no significant difference between the 2 groups for age, indication for surgery, pre-operative deformity or grade of the operating surgeon. There was no significant difference in the mean time immobilised, mean time to union or complications. The patients were interviewed by telephone after a mean follow-up of 9.4 months. There was no significant difference in results between the 2 groups. This suggests that a forefoot plaster following Mitchell's osteotomy is unnecessary. Postoperative mobilisation in a wooden soled shoe can be used as an alternative. PMID- 11890626 TI - Increased vascularisation enhances axonal regeneration within an acellular nerve conduit. AB - Despite major advances in microsurgical techniques, the functional results of periphery nerve repair remain largely unsatisfactory. Furthermore, if a defect exists such that a nerve graft is required, the results are generally worse than those following a primary repair. The autologous nerve graft, the current 'gold standard', has inherent disadvantages and there has been a long quest to find a suitable alternative. The role of vascularisation in nerve regeneration is poorly defined and the aim of this work was to define and quantify the effects of increased vascularisation on nerve regeneration. Rat sciatic nerve defects (1 cm) were bridged with a silicone chamber containing vascular endothelial growth factor (VEGF; 500 or 700 ng/ml) in a laminin-based gel (Matrigel) known to support axonal regeneration. Chambers were harvested between 5 and 180 days to follow the progression of neural and vascular elements. Following immunohistochemical staining, computerised image analysis demonstrated that VEGF significantly increased vascular, Schwann cell and axonal regeneration within the chamber up to 30 days post-insertion, and stimulated regeneration of up to 78% more myelinated axons at 180 days, compared to plain Matrigel control. Furthermore, the non-linear vascular dose-response to VEGF was clearly reflected in the Schwann cell and axonal staining intensity, supporting the highly significant relationship between vascularisation and Schwann cell staining seen within the chamber (P <0.001). Target-organ re-innervation at 180 days was similarly enhanced by VEGF in an identical dose-dependant manner. VEGF at 500 ng/ml increased recovery of gastrocnemius muscle weight by 17% and footpad innervation by 51% (P <0.05) compared to control, indicating the long-term functional benefits of VEGF. PMID- 11890627 TI - Are we really as good as we think we are? AB - Differences are examined in assessment and self-assessment scores, in oral and maxillofacial surgery trainees and MSc postgraduates, following the surgical removal of lower third molar teeth. This study found evidence of a surprising and worrying over-rating of their own surgical skills by many trainees and postgraduates. PMID- 11890629 TI - Continuous lumbar drainage. PMID- 11890628 TI - Femoral embolectomy. PMID- 11890630 TI - Reduction of a dislocated low contact stress rotating platform using a single AO screw. PMID- 11890631 TI - Predictive clinical and laboratory factors in the diagnosis of temporal arteritis. AB - Surgeons are frequently called upon to perform temporal artery biopsy in patients suspected of having temporal arteritis. In this study, we have attempted to identify clinical and laboratory features that may predict the results of temporal artery biopsy for the diagnosis of temporal arteritis. The medical records of patients undergoing temporal artery biopsy over a 10-year period in one hospital were reviewed. Details of presenting features were recorded and comparisons made between biopsy-positive and biopsy-negative patients. Of 59 patients who underwent temporal artery biopsy, the records of 51 patients were located. Of these, 17 patients had positive biopsy specimens and 33 had negative biopsies. In one patient, no temporal artery was found in the biopsy specimen. In the biopsy-positive patients, 69% had an erythrocyte sedimentation rate of greater than 50 mm/h compared to 31% of biopsy negative patients (P = 0.03). With regard to the other clinical and laboratory parameters that were evaluated, no statistically significant differences were found between biopsy-positive and biopsy-negative patients. PMID- 11890632 TI - Validation of skinfold thickness and hand-held impedance measurements for estimation of body fat percentage among Singaporean Chinese, Malay and Indian subjects. AB - Body fat percentage (BF%) was measured in 298 Singaporean Chinese, Malay and Indian men and women using a chemical four-compartment model consisting of fat, water, protein and mineral (BF%4C). In addition, weight, height, skinfold thickness and segmental impedance (from hand to hand) was measured. Body fat percentage was predicted using prediction equations from the literature (for skinfolds BF%SKFD) and using the manufacturer's software for the hand-held impedance analyser (BF%IMP). The subjects ranged in age from 18-70 years and in body mass index from 16.0 to 40.2 kg/m2. Body fat ranged from 6.5 to 53.3%. The biases for skinfold prediction (BF%4C-BF%SKFD, mean +/- SD) were -0.4+/-3.9, 2.3+/-4.1 and 3.1+/-4.2 in Chinese, Malay and Indian women, respectively, the Chinese being different from the Malays and Indians. The differences were significant from zero (P < 0.05) in the Malays and Indians. For the men, the biases were 0.5+/-3.8, 0.0+/-4.8 and 0.9+/-4.0 in Chinese, Malays and Indians, respectively. These biases were not significantly different from zero and not different among the ethnic groups. The biases for hand-held impedance BF% were 0.7+/-4.5, 1.5+/-4.4 and 0.4+/-3.8 in Chinese, Malay and Indian women. These biases were not significantly different from zero but the bias in the Chinese was significantly different from the biases in the Malays and Indians. In the Chinese, Malay and Indian men, the biases of BF%IMP were 0.7+/-4.6, 1.9+/-4.8 and 2.0+/-4.4, respectively. These biases in Malay and Indian men were significantly different from zero and significantly different from the bias in Chinese men. The biases were correlated with level of body fat and age, and also with relative arm span (arm span/height) for impedance. After correction, the differences in bias among the ethnic groups disappeared. The study shows that the biases in predicted BF% differ between ethnic groups, differences that can be explained by differences in body composition and differences in body build. This information is important and should be taken into account when comparing body composition across ethnic groups using predictive methods. PMID- 11890633 TI - Dietary patterns and nutrient intake of adult women in south-east China: a nutrition study in Zhejiang province. AB - This study documents the dietary patterns and nutrient intake of 652 adult women living in south-east China. Compared with data from previous national surveys and other nutrition studies in China, the results show different dietary patterns. The major differences include a greater consumption of vegetables, fruits and animal foods, but a lower consumption of cereal and tuber foods. The mean daily nutrient intakes of the urban women met the Chinese recommended dietary allowances. However, the situation was different in rural areas, where women had lower mean intakes of vitamins and minerals. There were also significant differences in dietary pattern and food consumption between these two groups of women. Further improvements in dietary intake for those residing in the south east rural areas of China are needed. PMID- 11890635 TI - Physicochemical properties and nutritional traits of millet-based weaning food suitable for infants of the Kumaon hills, Northern India. AB - A weaning food based on malted foxtail millet flour (30%), malted barnyard millet flour (30%), roasted soybean flour (25%) and skim milk powder (15%) was prepared. The mix contained 18.37 g protein and 398 kcal energy per 100g. The nutrient composition of this unfortified weaning (UW) mix met the Prevention of Food Adulteration (PFA) standards, except in total ash. In order to meet the minor constituent requirements, the UW mix was fortified. The fortified weaning (FW) mix met PFA standards for various nutrients. The protein efficiency ratio of the UW mix was 2.25 against a casein control, for which a value of 2.50 was recorded. The nutrient composition, viscosity and sensory quality of the UW mix was compared with the marketed weaning mix, commercial infant formula. The viscosity of UW gruel was much lower (20 centipoise (cps)) than that of marketed weaning mix (7400 cps). The high alpha-amylase activity of 661 units in the UW mix was responsible for its low viscosity. The sensory quality of UW mix and marketed weaning mix did not differ significantly (P = 0.05). Both of the gruels were liked moderately on the Hedonic Scale. The UW gruel met the acceptability criteria for weaning food. It could be stored in plastic airtight containers at room temperature for 4 months without any changes in sensory quality. PMID- 11890634 TI - Serum concentrations of micronutrient antioxidants in an adult Arab population. AB - Serum concentrations of retinol, alpha-tocopherol, beta-carotene and lycopene were measured by reversed-phase high-performance liquid chromatography (r-P HPLC) in 260 randomly selected healthy adult Kuwaitis (159 men and 101 women) aged 18 63 years (mean 33.3 years) to established reference ranges of the micronutrient antioxidants. Total cholesterol concentrations were assayed by an enzymatic method to determine alpha-tocopherol: cholesterol ratios. The mean +/- SEM (micromol/L) for retinol, alpha-tocopherol, beta-carotene and lycopene were 1.76+/-0.02, 20.0+/-0.5, 0.52+/-0.03, 0.95+/-0.05, respectively. Compared to other populations, these data showed, on the whole, ordinary concentrations of beta-carotene, comparatively low concentrations of retinol and alpha-tocopherol and high concentrations of lycopene. Retinol concentrations were similar for both sexes, whereas alpha-tocopherol concentration was significantly (P < 0.0001) lower and the carotenoid levels (beta-carotene and lycopene) significantly higher (P < 0.0001) in women. Of the micronutrient antioxidants, alpha-tocopherol was most correlated with cholesterol (r = 0.492, P < 0.0001). beta-Carotene and lycopene were highly correlated with each other (r =0.744, P< 0.0001). Age was positively associated with beta-carotene (r = 0.214, P = 0.001) and lycopene (r = 239, P< 0.0001). Our data enabled us to establish a gender non-specific reference range for retinol and gender-specific reference ranges for alpha-tocopherol, beta carotene and lycopene. PMID- 11890636 TI - Assessment of iodine deficiency in Kottayam district, Kerala State: a pilot study. AB - Iodine is one of the essential micro-elements required for normal human growth and development. Iodine Deficiency Disorders (IDD) are an important public health problem in India. There has been no data on the prevalence of IDD from the Kottayam district, India and hence, the present pilot study was conducted in the year 1999 to assess whether iodine deficiency existed in the district or not and to estimate the iodine content of salt consumed by the population. A total of 1872 children in the age group of 6-12 years were included in the study and were clinically examined. On the spot urine samples were collected from 251 children. A total of 420 salt samples were collected randomly from the families of the children. The total goitre prevalence was found to be 7.05% in the subjects studied. It was found that the percentage of children with urinary iodine excretions of < 2, 2- < 5, 5-9 and 10 microg/dL and above were 6.4%, 6.0%, 20.7%, and 66.9%, respectively. Assessment of the iodine content of salt by the iodometric titration method revealed that 60.6% of the children were consuming salt with an iodine content of 15 p.p.m. and more, which was the stipulated level of salt iodisation. The findings of the present study indicated that the population is in a transitional phase from iodine deficient, as revealed by total goitre rate, to iodine sufficient nutriture, as revealed by the median urinary iodine excretion level of 17.5 microg/dL. PMID- 11890638 TI - A prospective study of weight and height going from infancy to adolescence. AB - Weight and height from infancy to age 15 years was studied in the Geelong population (n = 1200 in infancy; n = 213 at adolescence), Victoria, Australia. Body mass index (BMI) increased from 3 months to 12 months and then decreased again until 80 months after which it increased to 20.5 kg/m2 at the age of 15 years. The extent of tracking of BMI in infants classified as overweight or underweight was similar and differed from that of subjects of normal weight. Only one in four of the infants classified as overweight or underweight in infancy were still in the same category in adolescence, compared with three in four of those classified as of normal weight. Socioeconomic status has an effect on weight and height status in adolescence but not on the tracking of BMI. The age at 6-7 years is a critical age for weight and height status in adolescence. It appears that weight and height in infancy have a significant relationship with body size in adolescence but only in boys. PMID- 11890637 TI - Will iron supplementation given during menstruation improve iron status better than weekly supplementation? AB - To investigate the efficacy of two different iron supplements administered either on a weekly basis or during menstruation, a 16-week community experimental study was carried out among postmenarcheal female adolescent students in Kupang, East Nusa Tenggara, Indonesia. Forty eight students received a placebo tablet weekly, 48 other students got an iron tablet weekly and 41 students took an iron tablet for four consecutive days during their menstruation cycle. All subjects were given deworming tablets before supplementation. Haemoglobin, serum ferritin, height, weight, mid-upper arm circumference and dietary intake were assessed before and after intervention. The supplementation contributed to a significant improvement in the iron status of the intervention groups compared to the placebo group (P < 0.05). In the menstruation group, the haemoglobin concentrations of the anaemic subjects improved significantly (P < 0.05) while for the non-anaemic subjects, serum ferritin concentrations also increased significantly (P < 0.05). In the weekly group for anaemic and nonanaemic subjects, there was a significant increase in both haemoglobin and serum ferritin concentrations (P < 0.05). This study revealed that weekly supplementation of iron tablets continued for 16 weeks contributed a higher improvement to haemoglobin concentration, compared with supplementing iron tablets for four consecutive days during menstruation for four menstrual cycles. This suggests that weekly iron supplementation is preferable. PMID- 11890639 TI - Role of plant metabolites in toxic liver injury. AB - Aphanamixis polystachya is a traditional medicinal plant of the Meliaceae family in India. A crude ethanolic extract of the leaf of this plant shows a beneficial effect on toxic liver injury. Its antihepatotoxic activity was evaluated on carbon tetrachloride (CCl4)-induced liver injury in a rat model. The assessment of hepatoprotective activity was evaluated by measuring the activities of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH), serum total bilirubin and albumin and histology of the liver. The crude leaf extract significantly inhibits the enhanced ASAT, ALAT, ALP, ACP and LDH activities released from the CCl4-intoxicated animals. It also ameliorated the depressed value of serum albumin and the enhanced value of total bilirubin in plasma caused by CCl4 intoxication. The study showed that the crude ethanolic extract from A. polystachya leaves provided protection against acute carbon tetrachloride-induced liver damage. PMID- 11890640 TI - Characteristics of soy bread users and their beliefs about soy products. AB - A two-stage random telephone/mail survey was conducted during the last quarter of 1998 among Adelaide residents to determine consumers' use of soy bread and other soy products and their health expectations of soy products. One in five (21%) of 1477 telephone subscribers usually consumed soy bread and related soy products. Comparisons of soy bread consumers and non-consumers, based on the mail survey sample, showed that more soy bread consumers used dietary supplements and ate low fat and vegetarian diets, though their experiences of ill health were similar. Soy bread consumers held stronger universalism (pro-nature) values than non consumers. They also held more positive expectations about the benefits of soy consumption, including reductions in menstrual and menopausal symptoms, increased bowel regularity and reductions in the risk of heart disease and cancer. The findings are discussed in relation to the psychology of dietary supplementation, values orientations and physiological plausibility. Further investigations are suggested. PMID- 11890641 TI - Association of overall and abdominal obesity with coronary heart disease risk factors: comparison between urban and rural Indian men. AB - The relationship of body mass index (BMI), conicity index (CI) and waist circumference to four coronary heart disease (CHD) risk factors (systolic and diastolic blood pressures, total cholesterol and high-density lipoprotein (HDL) cholesterol levels) was examined in urban (n = 110) and rural (n = 102) men aged > or = 20 years, drawn from the 'Reddy' population of Southern Andhra Pradesh, India. Using ANCOVA we found significant difference (< 0.01) for systolic blood pressure, total cholesterol and HDL cholesterol between the urban and rural samples. The Pearson's correlation coefficients suggest that BMI and waist circumference had significant relationships with most of the risk factors in both the populations. The CI did not significantly influence any of the risk factors in the urban population; however, in the rural population, CI did show a significant positive relationship with both of the blood pressures and with TC. Even after controlling for age, smoking and physical activity (partial correlations), the relations remained constant. In multiple linear regression, BMI showed significant positive association with systolic and diastolic blood pressures (<0.01) and HDL cholesterol (<0.05) in the rural population only. However, the Cl showed a significant association with HDL cholesterol, and waist circumference with total cholesterol and HDL cholesterol in the rural population. The results of the present study revealed that BMI and waist circumference had a greater influence on the CHD risk factors, and that the influence was more conspicuous in the rural sample. Comparing the association of abdominal obesity measures (CI and waist circumference) with CHD risk factors, waist circumference better correlated with most of the risk factors. Hence the present study suggests that BMI and waist circumference are better indicators of CHD risk factors. However, the importance of Cl has to be further studied in South Asian populations. PMID- 11890642 TI - Cholesterol oxides: their occurrence and methods to prevent their generation in foods. AB - Eight cholesterol oxides are commonly found in foods with high cholesterol content, such as meat, egg yolk and full fat dairy products. Factors known to increase the production of cholesterol oxides in foods are heat, light, radiation, oxygen, moisture, low pH, certain pro-oxidising agents and the storage of food at room temperature. Processes, such as pre-cooking, freeze-drying, dehydration and irradiation, have all been reported to result in increased production of cholesterol oxides in meats. As prepared consumer foods are becoming increasingly popular, the consumption of higher levels of cholesterol oxides in foods is inevitable. An understanding of the mechanisms involved in the generation of cholesterol oxides may assist in their reduction in foods and possibly reduce the impact of these compounds on human health. PMID- 11890643 TI - Anticancer and health protective properties of citrus fruit components. AB - Accumulated evidence from experimental and epidemiological studies indicates that there is a low risk of degenerative diseases, cardiovascular disease, hypertension, cataract, stroke and, in particular, cancers in people with a high intake of fruit and vegetables. This protective effect is assumed to be associated mainly with the antioxidant activities of either individual or interacting bioactive components present in the fruits and vegetables, and with other biochemical and physical characteristics of the identified and unknown bioactive components. The implicated bioactive components present in citrus fruits include vitamin C, beta-carotene, flavonoids, limonoids, folic acid, and dietary fibre. A high intake of citrus fruits may reduce the risk of degenerative diseases. PMID- 11890644 TI - Dietary patterns and risk factors for type 2 diabetes mellitus in Fijian, Japanese and Vietnamese populations. AB - Diabetes mellitus is now a serious and increasing problem in Asian countries, where dietary patterns have shifted toward Westernized foods and people are becoming more sedentary. In order to elucidate the relationship of dietary habits to the development of diabetic risk factors, the dietary patterns of 200 Fijian, 171 Japanese and 181 Vietnamese women of 30-39 years of age were investigated using 3 day-24 h recall or dietary records. Anthropometric measurements and glycosuria tests were also conducted. The dietary trends of Fijians and Japanese have changed drastically in the past 50 years, while Vietnamese have been minimally influenced by Western dietary habits. The mean 24 h dietary intake showed that Fijians had the highest energy intake. Energy intake from fat was only 13% for Vietnamese, but over 30% for Japanese and Fijians. Percentage of body fat was higher in Vietnamese than in Japanese, though there were no significant differences in body mass index (BMI). In the overweight and obese women, Vietnamese had higher abdominal obesity than Japanese. The prevalence of obesity (BMI > or = 30 kg/m2) was 63.0% for Fijians, 1.8% for Japanese and 1.1% for Vietnamese. Glycosuria testing yielded the most positive cases among Fijians. Dietary transition and dietary excess appear to be potential risk factors for diabetes in Fijian women. PMID- 11890645 TI - High efficiency gene transfer is an efficient way of defining therapeutic targets: a functional genomics approach. AB - BACKGROUND: Dendritic cells are the most potent antigen presenting cells and key to many aspects of the immune function. Studying the intracellular signalling mechanism used by dendritic cells would provide an insight into the functioning of these cells and give clues to strategies for immunomodulation. METHOD: Highly efficient adenoviral infection of dendritic cells for the delivery of transgenes was obtained. These viral vectors were used to introduce IkappaB alpha into dendritic cells for the inhibition of NF-kappaB. This was used to investigate the role of NF-kappaB in dendritic cell function. RESULTS: By blocking the NF-kappaB function a potent inhibition of the expression of costimulating molecules by dendritic cells with the concomitant loss of T cell stimulating function was demonstrated. CONCLUSION: The use of adenoviral vectors may be a useful way of studying the role of genes in dendritic cell function. PMID- 11890647 TI - Updated consensus statement on tumour necrosis factor blocking agents for the treatment of rheumatoid arthritis and other rheumatic diseases (April 2001). AB - TNF blocking agents have proved to be effective DMARDs and they have been a major advance in the treatment of RA. Their use is expanding to other rheumatic diseases. However, rare to uncommon and unexpected toxicities have been found and others may yet be found during their use. Studies in selected areas of efficacy, toxicity, and general use of TNF blocking agents are needed to help further define the most appropriate use of these agents. Use of these drugs will require doctors experienced in the diagnosis, treatment, and assessment of RA and other rheumatic diseases. These doctors will need to make long term observations of efficacy and toxicity. Further considerations which must be made when using TNF blocking agents in this disease include the cost and a recognition that data in subgroups are still being acquired. It is hoped that this statement, which is based upon the best evidence available at the time of its creation, and modified by expert opinion, will facilitate the optimal use of these agents for our patients with RA and other rheumatic diseases. PMID- 11890646 TI - Novel targets for interleukin 18 binding protein. AB - BACKGROUND: Interleukin 18 (IL18) is related to the IL1 family by structure, receptors, signalling molecules, and function. IL18 induces gene expression and synthesis of tumour necrosis factor (TNF), IL1, Fas ligand, several chemokines, and vascular adhesion molecules. Similar to IL1beta, IL18 is synthesised as a biologically inactive precursor molecule lacking a signal peptide. The IL18 precursor requires cleavage into an active, mature molecule by the intracellular cysteine protease, IL1beta converting enzyme (ICE, or caspase-1). Inhibitors of ICE activity limit the biological activity of IL18 in animals and may be useful in reducing the activity of IL18 in human disease. However, a constitutively secreted IL18 binding protein (IL18BP) exists which functions as a natural inhibitor of IL18 activity. IL18BP binds IL18 with a high affinity (Kd of 400 pM) and, at equimolar ratios, inhibits 50-70% of IL18; at twofold molar excess, IL18BP neutralises nearly all IL18 activity. METHOD: IL18 was investigated for its role in human myocardial function. An ischaemia/reperfusion (I/R) model of suprafused human atrial myocardium was used to assess myocardial contractile force. RESULTS: The addition of IL18BP to the perfusate during and after I/R resulted in improved post-I/R contractile function from 35% of control to 76% with IL18BP. Also, IL18BP treatment preserved intracellular tissue creatine kinase levels (by 420%). Because active IL18 requires cleavage of its precursor form by ICE, inhibition of ICE attenuated the depression in contractile force after I/R (from 35% of control compared with 75.8% in treated atrial muscle, p<0.01). CONCLUSION: Myocardial ischaemia is a target for IL18BP and use of IL18BP may thereby reduce ischaemia-induced myocardial dysfunction. PMID- 11890648 TI - Biology of TACE inhibition. AB - Studies conducted over the past decade have demonstrated a central role for tumour necrosis factor alpha (TNFalpha) in inflammatory diseases. As a result of this work, a number of biological agents that neutralise the activity of this cytokine have entered the clinic. The recent clinical data obtained with etanercept and infliximab highlight the relevance of this strategy. TNFalpha converting enzyme (TACE) is the metalloproteinase that processes the 26 kDa membrane bound precursor of TNFalpha (proTNFalpha) to the 17 kDa soluble component. Although a number of proteases have been shown to process proTNFalpha, none do so with the efficiency of TACE. A series of orally bioavailable, selective, and potent TACE inhibitors are currently in clinical development. These inhibitors effectively block TACE mediated processing of proTNFalpha and can reduce TNF production by lipopolysaccharide stimulated whole blood by >95%. Through a series of studies it is shown here that >80% of the unprocessed proTNFalpha is degraded intracellularly. The remainder appears to be transiently expressed on the cell surface. Although, in vitro, TACE inhibition has also been implicated in shedding of p55 and p75 surface TNFalpha receptors, the in vivo data cast doubt on the consequences of this finding. In a mouse model of collagen induced arthritis, the inhibitors are efficacious both prophylactically and therapeutically. The efficacy seen is equivalent to strategies that neutralise TNFalpha. In many studies greater efficacy is observed with the TACE inhibitors, presumably owing to greater penetration to the site of TNFalpha production. PMID- 11890649 TI - Treatment of active spondyloarthropathy with infliximab, the chimeric monoclonal antibody to tumour necrosis factor alpha. PMID- 11890651 TI - Treatment of myelodysplastic syndrome with agents interfering with inhibitory cytokines. AB - Results of these trials provide evidence for biological activity and some clinical efficacy of agents potentially blocking inhibitory cytokines in patients with MDS. However, given the limited responses, it appears that factors additional to TNFalpha inhibitory activity contribute to the development of cytopenias in these patients. Further studies are warranted using anti TNFalpha/anti-inhibitory cytokine approaches, either alone or in combination with other agents, capable of abrogating the effects of additional inhibitory mechanisms in MDS. PMID- 11890650 TI - Cytokine blockers in psoriatic arthritis. AB - The cellular events underlying the pathogenesis of psoriatic arthritis (PsA) and psoriasis have not yet been fully elucidated. Nevertheless, some clues to these conditions are beginning to emerge. In particular, a growing body of data supports the role of proinflammatory cytokines, such as tumour necrosis factor (TNF), in the pathophysiology of PsA and psoriasis. Raised levels of these cytokines are found in the joints of patients with PsA, as well as in psoriatic skin lesions. Physiotherapy, non-steroidal anti-inflammatory agents, corticosteroids, and disease modifying antirheumatic agents, such as methotrexate, are the most commonly used treatments for PsA. However, the data supporting the effectiveness of these treatments are limited, and disease resolution is usually incomplete. This study examined the effects of etanercept, a TNF inhibitor, in patients with PsA. Etanercept treatment was well tolerated and resulted in significant improvement in the signs and symptoms of PsA and in psoriatic skin lesions. Infliximab, another TNF inhibitor, has also been shown to be effective in patients with PsA. Such studies confirm the importance of proinflammatory cytokines in PsA, and hold out hope for patients who require new options for the treatment of their disease. PMID- 11890652 TI - Role of cytokines in the innate immune response to intracellular pathogens. PMID- 11890653 TI - Radiographic progression in rheumatoid arthritis: does it reflect outcome? Does it reflect treatment? AB - Although there is clear face validity that structural damage is related to outcome, it is difficult to prove this. Recently, more evidence became available that structural damage, as assessed on plain films, is indeed related to disease activity in clinical trials and therefore can be used to assess the effect of treatment. Also, a relationship between structural damage and outcome, mainly defined as physical disability, was established. Several examples of findings in recent publications are presented which lead to the following conclusions. There is a relation between the response to treatment measured as clinical disease activity and measured as radiographic progression in most clinical therapeutic trials. A strong relation between local inflammation and progression of damage in the individual joint is present. This is robust evidence for the hypothesis that inflammation leads to structural damage. There is a good relation between the damage in small and large joints as assessed on plain films. Damage measured in small joints is a good substitute for overall damage. Disease activity is always strongly correlated with functional disability throughout the disease course. There is an increasing relation between disability and structural damage with increasing disease duration. PMID- 11890654 TI - "Stepping-up" from methotrexate: a systematic review of randomised placebo controlled trials in patients with rheumatoid arthritis with an incomplete response to methotrexate. PMID- 11890655 TI - Successful treatment of a small cohort of patients with adult onset of Still's disease with infliximab: first experiences. AB - OBJECTIVE: To test the efficacy of infliximab in the treatment of patients with severe and active adult onset Still's syndrome (AOSD) despite conventional immunosuppressive therapy. PATIENTS AND METHODS: Six patients with the diagnosis of AOSD according to the Yamagushi criteria of 1992 were treated with infliximab. All patients had severe disease with high clinical and serological activity. Patients were treated initially with high dose steroids or more intensive immunosuppressive therapy. Two patients had a history of multiple disease modifying antirheumatic drug (DMARD) treatments. One patient had a history of three years of AOSD with fever, chills, pleural and pericardial effusions, and hepatosplenomegaly. Despite these treatments, he developed increasing serological signs of inflammation and arthritis of both hips and peripheral joints. Another patient had a history of five years of AOSD with oligoarthritis, myalgias, and recurrent fever despite multiple DMARD treatment, including cyclophosphamide pulse therapy. Our patients with AOSD presented with massive polyarthralgias, polyarthritis, splenomegaly or hepatomegaly, the typical rash, sore throat, weight loss, serositis, continuing fever, leucocytosis, and raised C reactive protein (CRP), erythrocyte sedimentation rate (ESR), and ferritin levels. Four patients with early onset of the disease, fulfilling the diagnostic criteria for AOSD and a clinical and serological high disease activity, were included in our pilot study without any further DMARD treatment apart from the initial steroid treatment. Reduction of established treatment, mainly with steroids, caused a relapse of the disease in all our patients. Patients then received 3-5 mg/kg infliximab on weeks 0, 2, and 6, continuing with intervals of 6-8 weeks depending on the patient's individual disease activity. RESULTS: In all patients, fever, arthralgias, myalgias, hepatosplenomegaly, and the rash resolved after the first courses of treatment with infliximab. All serological variables (CRP, ESR, hyperferritinaemia) returned to normal. After three courses of infliximab infusions, splenomegaly could not be detected in any of our patients. One caused by hip postarthritic osteoarthrosis, requiring hip replacement. After three courses of treatment with infliximab, splenomegaly could not be detected in any of our patients. Up to now, our patients have received infliximab infusion treatment for between five and 28 months. Throughout this period all patients have continued to benefit from this treatment, with improvement in their clinical symptoms, joint counts, and serological disease activity. One of our patients had a moderate infusion reaction during the second treatment. The infusion was discontinued for one hour and then was resumed with no further problems. CONCLUSION: The disease improved remarkably in all six patients with AOSD after treatment with infliximab, also in the early stage of AOSD. These preliminary data suggest the potential therapeutic benefit of anti-tumour necrosis factor alpha treatment in AOSD. PMID- 11890656 TI - New treatment options in ankylosing spondylitis: a role for anti-TNFalpha therapy. AB - Anti-TNFalpha treatment seems to be highly effective in AS. The results available indicate that this treatment is at least as effective as in RA. Furthermore, because no other treatments are available for AS--in contrast with RA or psoriatic arthritis--infliximab may even become a first line immunosuppressive treatment in patients with severe active AS. A dose of 5 mg/kg body weight seems to be required and intervals between 6 and 12 weeks seem to be necessary depending on the disease activity. It remains to be seen what the long term effects will be, whether the patients benefit from long term treatment, and whether radiological progression and ankylosis can be stopped. Allergy, lupus like diseases, and tuberculosis are rare side effects which need to be considered. At first glance, the possible benefits of anti-TNFalpha treatment seem to outweigh these shortcomings, including the high cost. PMID- 11890658 TI - Matrix metalloproteinase inhibitors in rheumatic diseases. AB - The rheumatic diseases continue to represent a significant healthcare burden in the 21st century. However, despite the best standard of care and recent therapeutic advances it is still not possible to consistently prevent the progressive joint destruction that leads to chronic disability. In rheumatoid arthritis and osteoarthritis this progressive cartilage and bone destruction is considered to be driven by an excess of the matrix metalloproteinase (MMP) enzymes. Consequently, a great number of potent small molecule MMP inhibitors have been examined. Several MMP inhibitors have entered clinical trials as a result of impressive data in animal models, although only one MMP inhibitor, Ro32 3555 (Trocade), a collagenase selective inhibitor, has been fully tested in the clinic, but it did not prevent progression of joint damage in patients with rheumatoid arthritis. The key stages and challenges associated with the development of an MMP inhibitor in the rheumatic diseases are presented below with particular reference to Trocade. It is concluded that the future success of MMP inhibitors necessitates a greater understanding of the joint destructive process and it is hoped that their development may be accompanied with clearer, more practical, outcome measures to test these drugs for, what remains, an unmet medical need. PMID- 11890657 TI - Differential roles of Toll-like receptors in the elicitation of proinflammatory responses by macrophages. AB - BACKGROUND: Mammalian Toll-like receptor (TLR) proteins are pattern recognition receptors for a diverse array of bacterial and viral products. Gram negative bacterial lipopolysaccharide (LPS) activates cells through TLR4, whereas the mycobacterial cell wall glycolipids, lipoarabinomannan (LAM) and mannosylated phosphatidylinositol (PIM), activate cells through TLR2. Furthermore, short term culture filtrates of M. tuberculosis bacilli contain a TLR2 agonist activity, termed soluble tuberculosis factor (STF), that appears to be PIM. It was recently shown that stimulation of RAW264.7 murine macrophages by LPS, LAM, STF, and PIM rapidly activated NF-kappaB, AP1, and MAP kinases. RESULTS: This study shows that signalling by TLR2 and TLR4 also activates the protein kinase Akt, a downstream target of phosphatidylinositol-3'-kinase (PI-3-K). This finding suggests that activation of PI-3-K represents an additional signalling pathway induced by engagement of TLR2 and TLR4. Subsequently, the functional responses induced by the different TLR agonists were compared. LPS, the mycobacterial glycolipids, and the OspC lipoprotein (a TLR2 agonist) all induced macrophages to secrete tumour necrosis factor alpha (TNFalpha), whereas only LPS could induce nitric oxide (NO) secretion. Human alveolar macrophages also exhibited a distinct pattern of cellular response after stimulation with TLR2 and TLR4 agonists. Specifically, LPS induced TNFalpha, MIP-1beta, and RANTES production in these cells, whereas the TLR2 agonists induced only MIP-1beta production. CONCLUSION: Together, these data show that different TLR proteins mediate the activation of distinct cellular responses, despite their shared ability to activate NF-kappaB, AP1, MAP kinases, and PI-3-K. PMID- 11890659 TI - The atrial natriuretic peptide regulates the production of inflammatory mediators in macrophages. AB - The atrial natriuretic peptide (ANP), a member of the natriuretic peptide family, is a cardiovascular hormone which possesses well defined natriuretic, diuretic, and vasodilating properties. Most of the biological effects of ANP aremediated through its guanylyl cyclase coupled A receptor. Because ANP and its receptors have been shown to be expressed and differentially regulated in the immune system, it has been suggested that ANP has an immunomodulatory potency. Much investigation of the effects of ANP on the activation of macrophages has been carried out. ANP was shown to inhibit the lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS) in macrophages in an autocrine fashion. ANP in this context was shown to reduce significantly the activation of NF-kappaB and to destabilise iNOS mRNA. ANP, furthermore, can significantly reduce the LPS-induced secretion of tumour necrosis factor alpha (TNFalpha) in macrophages. The relevance of these findings on a regulatory role for ANP on TNFalpha in humans was shown by the fact that ANP significantly reduces the release of TNFalpha in whole human blood. It was furthermore shown to attenuate the release of interleukin 1beta (IL1beta). Interestingly, ANP did not affect the secretion of the anti-inflammatory cytokines IL10 and IL1 receptor antagonist (IL1ra). In summary, ANP was shown to reduce the secretion of inflammatory mediators in macrophages. Therefore, this cardiovascular hormone may possess anti-inflammatory potential. PMID- 11890661 TI - Oncostatin M in the anti-inflammatory response. AB - Oncostatin M (OM) is a pleiotropic cytokine of the interleukin 6 family, whose in vivo properties and physiological function remain in dispute and poorly defined. These in vivo studies strongly suggest that OM is anabolic, promoting wound healing and bone formation, and anti-inflammatory. In models of inflammation OM is produced late in the cytokine response and protects from lipopolysaccharide (LPS)-induced toxicities, promoting the re-establishment of homoeostasis by cooperating with proinflammatory cytokines and acute phase molecules to alter and attenuate the inflammatory response. Administration of OM inhibited bacterial LPS induced production of tumour necrosis factor alpha and septic lethality in a dose dependent manner. Consistent with these findings, in animal models of chronic inflammatory disease OM potently suppressed inflammation and tissue destruction in murine models of rheumatoid arthritis and multiple sclerosis. T cell function and antibody production were not impaired by OM treatment. Taken together, these data indicate that the activities of this cytokine in vivo are anti-inflammatory without concordant immunosuppression. PMID- 11890660 TI - Antagonising angiogenesis in rheumatoid arthritis. PMID- 11890663 TI - Bone tissue engineering: hope vs hype. AB - The requirement for new bone to replace or restore the function of traumatised, damaged, or lost bone is a major clinical and socioeconomic need. Bone formation strategies, although attractive, have yet to yield functional and mechanically competent bone. Bone tissue engineering has been heralded as the alternative strategy to regenerate bone. In essence, the discipline aims to combine progenitor or mature cells with biocompatible materials or scaffolds, with or without appropriate growth factors, to initiate repair and regeneration. This brief review outlines the concepts, challenges, and limitations in bone tissue engineering and the potential that could improve the quality of life for many as a result of interdisciplinary collaboration. PMID- 11890662 TI - Osteoprotegerin. PMID- 11890664 TI - Nuclear translocation of SHP and visualization of interaction with HNF-4alpha in living cells. AB - Mutations in small heterodimer partner (SHP) and hepatocyte nuclear factor 4alpha (HNF4alpha) are associated with mild obesity and diabetes mellitus, respectively. Both receptors work together to determine the normal pancreatic beta-cell function. We examined their subcellular localization and interaction in living cells by tagging them with yellow and cyan variants of green fluorescent protein (GFP) variants. Expressed SHP resided only in the cytoplasm in COS-7 cells which lacks HNF4alpha, but predominantly in the nucleus in insulinoma cells (MIN6). HNF4alpha was localized exclusively in the nuclei of both cells, coexpressed with HNF4alpha in COS-7 cells, redistributed in the nucleus, depending on the amount of HNF4alpha. We found fluorescence resonance energy transfer between GFP-tagged SHP and HNF4alpha, indicating a specific close association between them in the nucleus. The results strongly suggest that SHP exists primarily in the cytoplasm and is translocated into the nucleus on interacting with its nuclear receptor partner HNF4alpha. PMID- 11890665 TI - Localization of PTP-FERM in nerve processes through its FERM domain. AB - PTP-FERM is a protein tyrosine phosphatase (PTP) of Caenorhabditis elegans containing a FERM domain and a PDZ domain. Here we report the characterization of PTP-FERM and the essential role of its FERM domain in the localization of PTP FERM in the worm. There are at least three alternatively spliced PTP-FERM isoforms, all of which contain a band 4.1/FERM domain, a PDZ domain, and a catalytic domain. PTP-FERM possessed phosphatase activity. PTP-FERM was expressed predominantly in neurons in the nerve ring and the ventral nerve cord. PTP-FERM was found in the nerve processes and to be enriched in the peri-membrane region. Studies using various deletion mutants revealed that the FERM domain was essential and sufficient for the subcellular localization. These results suggest the essential role of the FERM domain in the function of PTP-FERM in the neurons of C. elegans. PMID- 11890666 TI - Immortalized suprachiasmatic nucleus cells express components of multiple circadian regulatory pathways. AB - We undertook an extensive antigenic characterization of the SCN 2.2 cell line in order to further evaluate whether the line expresses components of circadian regulatory pathways common to the hypothalamic suprachiasmatic nucleus (SCN), the central circadian clock in mammals. We found that differentiated SCN 2.2 cultures expressed a broad range of putative clock genes, as well as components of daytime, nighttime, and crepuscular circadian regulatory pathways found within the SCN in vivo. The line also exhibits several antigens that are highly expressed in a circadian pattern and/or differentially localized in the SCN relative to other hypothalamic regions. Expression of a broad complement of circadian regulatory proteins and putative clock genes further support growing evidence in recent reports that the SCN 2.2 cell line is an appropriate model for investigating the regulation of central mammalian pacemaker. PMID- 11890667 TI - Molecular aptamer beacons for real-time protein recognition. AB - One of the most pressing problems facing those attempting to understand the regulation of gene expression and translation is the necessity to monitor protein production in a variety of metabolic states. Thus far, there is no easy solution that will either identify or quantitate proteins in real time. Here we introduce a novel protein probe, molecular aptamer beacon (MAB), for real time protein recognition and quantitative analysis. The MAB combines the signal transduction mechanism of molecular beacons and the molecular recognition specificity of aptamers. An MAB based on a thrombin-binding aptamer was prepared as a model to demonstrate the feasibility. Significant fluorescent signal change was observed when MAB was bound to thrombin, which is attributed to a significant conformational change in MAB from a loose random coil to a compact unimolecular quadruplex. The MAB recognizes its target protein with high specificity and high sensitivity (112 picomolar thrombin concentration) in homogeneous solutions. Ratiometric imaging has been conducted with MAB labeled with two fluorophores, which makes it feasible for protein quantitation in living specimen. The unique properties of the MAB will enable the development of a class of protein probes for real time protein tracing in living specimen and for efficient biomedical diagnosis in homogeneous solutions. PMID- 11890668 TI - TNF receptor 1, IL-1 receptor, and iNOS genetic knockout mice are not protected from anthrax infection. AB - Anthrax produces at least two toxins that cause an intense systemic inflammatory response, edema, shock, and eventually death. The relative contributions of various elements of the immune response to mortality and course of disease progression are poorly understood. We hypothesized that knockout mice missing components of the immune system will have an altered response to infection. Parent strain mice and knockouts were challenged with LD95 of anthrax spores (5 x 10(6)) administered subcutaneously. Our results show that all genetic knockouts succumbed to anthrax infection at the same frequency as the parent. TNF antibody delayed death but TNF receptor 1 knockout had no effect. IL-1 receptor or iNOS knockouts died sooner. Anthrax was more abundant in the injection site of TNF alpha and iNOS knockouts compared to parent suggesting that attenuated cellular response increases rate of disease progression. With the exception of edema and necrosis at the injection site pathological changes in internal organs were not observed. PMID- 11890669 TI - Thioredoxin reductase reduces lipid hydroperoxides and spares alpha-tocopherol. AB - We investigated whether and how rat liver thioredoxin reductase spares alpha tocopherol in biomembranes. Purified hydroperoxides of beta-linoleoyl-gamma palmitoylphosphatidylcholine were decreased 35% by treatment with thioredoxin reductase and 54% by thioredoxin reductase plus E. coli thioredoxin. Thioredoxin reductase also halved the amount of hydroperoxides that had been formed during photoperoxidation of liposomes composed of beta-linoleoyl-gamma palmitoylphosphatidylcholine, and of emulsions of both cholesterol and cholesteryl linolenate. In erythrocyte ghosts, thioredoxin reductase spared alpha tocopherol from oxidation by both soybean lipoxygenase and ferricyanide. Thioredoxin reductase also decreased F(2)-isoprostanes in ghosts oxidized by ferricyanide, suggesting that its ability to spare alpha-tocopherol relates to reduction of lipid hydroperoxides. PMID- 11890671 TI - A double RING-H2 domain in RNF32, a gene expressed during sperm formation. AB - The RING domain is a cysteine-rich zinc-binding motif, which is found in a wide variety of proteins, among which are several proto-oncogenes and the gene implicated in autosomal recessive juvenile parkinsonism, Parkin. The domain mediates binding to other proteins, either via their RING domains or other motifs. In several proteins, RING domains are found in combination with other cysteine-rich binding motifs and some proteins contain two RING domains. Recent evidence suggests that RING finger proteins function in the ubiquitin pathway as E3 ligases. A variant of the RING domain is the RING-H2 domain, in which one of the cysteines is replaced by a histidine. We have cloned and characterized a novel gene, RNF32, located on chromosome 7q36. RNF32 is contained in 37 kb of genomic DNA and consists of 9 constitutive and 8 alternatively spliced exons, most of which are alternative first exons. A long and a short transcript of the gene are expressed; the short transcript containing exons 1-4 only. This gene encodes two RING-H2 domains separated by an IQ domain of unknown function. This is the first reported gene with a double RING-H2 domain. In humans, RNF32 overlaps with a processed retroposon located on the opposite strand, C7orf13. RNF32 is specifically expressed in testis and ovary, whereas C7orf13 is testis specific, suggesting that its expression may be regulated by elements in the RNF32 promoter region. RNF32 is expressed during spermatogenesis, most likely in spermatocytes and/or in spermatids, suggesting a possible role in sperm formation. PMID- 11890670 TI - Chemical induction of cellular antioxidants affords marked protection against oxidative injury in vascular smooth muscle cells. AB - Extensive evidence suggests that reactive oxygen species are critically involved in the pathogenesis of cardiovascular diseases, such as atherosclerosis and myocardial ischemia-reperfusion injury. Consistent with this concept, administration of exogenous antioxidants has been shown to be protective against oxidative cardiovascular injury. However, whether induction of endogenous antioxidants by chemical inducers in vasculature also affords protection against oxidative vascular cell injury has not been extensively investigated. In this study, using rat aortic smooth muscle A10 cells as an in vitro system, we have studied the induction of cellular antioxidants by the unique chemoprotector, 3H 1,2-dithiole-3-thione [corrected] (D3T) and the protective effects of the D3T induced cellular antioxidants against oxidative cell injury. Incubation of A10 cells with micromolar concentrations of D3T for 24 h resulted in a significant induction of a battery of cellular antioxidants in a concentration-dependent manner. These included reduced glutathione (GSH), GSH peroxidase, GSSG reductase, GSH S-transferase, superoxide dismutase, and catalase. To further examine the protective effects of the induced endogenous antioxidants against oxidative cell injury, A10 cells were pretreated with D3T and then exposed to either xanthine oxidase (XO)/xanthine, 4-hydroxynonenal, or cadmium. We observed that D3T pretreatment of A10 cells led to significant protection against the cytotoxicity induced by XO/xanthine, 4-hydroxynonenal or cadmium, as determined by 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium reduction assay. Taken together, this study demonstrates for the first time that a number of endogenous antioxidants in vascular smooth muscle cells can be induced by exposure to D3T, and that this chemical induction of cellular antioxidants is accompanied by markedly increased resistance to oxidative vascular cell injury. PMID- 11890672 TI - Active site mutations of recombinant deacetoxycephalosporin C synthase. AB - Site-directed mutagenesis of active site residues of deacetoxycephalosporin C synthase active site residues was carried out to investigate their role in catalysis. The following mutations were made and their effects on the conversion of 2-oxoglutarate and the oxidation of penicillin N or G were assessed: M180F, G299N, G300N, Y302S, Y302F/G300A, Y302E, Y302H, and N304A. The Y302S, Y302E, and Y302H mutations reduced 2-oxoglutarate conversions and abolished (<2%) penicillin G oxidation. The Y302F/G300A mutation caused partial uncoupling of penicillin G oxidation from 2-oxoglutarate conversion, but did not uncouple penicillin N oxidation from 2-oxoglutarate conversion. Met-180 is involved in binding 2 oxoglutarate, and the M180F mutation caused uncoupling of 2-oxoglutarate from penicillin oxidation. The N304A mutation apparently enhanced in vitro conversion of penicillin N but had little effect on the oxidation of penicillin G, under standard assay conditions. PMID- 11890673 TI - Sp1 and Sp3 activate the rat connexin40 proximal promoter. AB - The rat gap junction protein connexin40 (rCx40) has a characteristic developmental and regional expression pattern, for which the exact regulatory mechanisms are not known. To identify the molecular factors controlling Cx40 expression, its proximal promoter was characterized. The proximal rCx40 promoter is the most conserved noncoding region within the Cx40-gene known thus far and contains five potential binding sites for Sp-family transcription factors. The binding of both Sp1 and Sp3 to each of these DNA elements was demonstrated by EMSA. Luciferase assays of the natural rCx40 proximal promoter or mutated derivatives in Cx40-expressing (NCM, primary rat neonatal cardiomyocytes and A7r5, rat smooth muscle embryonic thoracic aorta cells) and -nonexpressing cells (N2A, mouse neuroblastoma cells) revealed that all sites are contributing to basal promoter activity. Trans-activation assays in Drosophila Schneider line 2 cells demonstrated that Sp1 and Sp3 activate the rCx40 proximal promoter in a dose-dependent and additive manner. PMID- 11890674 TI - Vascular endothelial growth factor induces IP-10 chemokine expression. AB - We have previously shown that intracavernous injection of vascular endothelial growth factor (VEGF) improved the recovery of erectile function in an arteriogenic impotence rat mode. We wished to identify genes that are affected by VEGF treatment in the penis. Specifically we examined the induction of IP-10 chemokine. Male rats were subjected to pudendal arterial ligation or sham operation. They were then treated with intracavernous injection of 4 microg of VEGF in phosphate-buffered saline (PBS) or PBS alone. At 6 and 24 h after treatment, the erectile tissue was then harvested for RNA isolation. The isolated RNA was used for microarray and RT-PCR analyses. Cultured rat cavernous smooth muscle cells (CSMC) were treated with VEGF and then subjected to RT-PCR analysis. Cultured human CSMC were treated with VEGF and then subjected to ELISA analysis. Microarray analysis detected IP-10 as an abundantly induced message in 6-h VEGF treated tissues. This was further confirmed by RT-PCR analysis. Using cultured rat CSMC, induction of IP-10 mRNA was detectable in 1 and 2 h, but not 24 h, VEGF treated cells. Induction of IP-10 at the protein level was observed with cultured human CSMC. Secretion of IP-10 into culture medium peaked at 4 h after treatment of human CSMC with 10 ng/ml of VEGF. Optimal VEGF dosage for IP-10 induction was 50 ng/ml when assayed with cells that were treated for 8 h. PMID- 11890675 TI - Apolipoprotein E activates Akt pathway in neuro-2a in an isoform-specific manner. AB - Apolipoprotein E (apoE) is a ligand for members of the low density lipoprotein (LDL) receptor family, receptors highly expressed in neurons. A study of one of the mechanisms by which apoE might affect neuronal cell metabolism is reported herein. ApoE can induce Akt/protein kinase B phosphorylation in Neuro-2a via two different pathways. Both pathways are mediated by phosphatidylinositol 3-kinase and cAMP-dependent protein kinase. The first pathway is stimulated by apoE3 and E4, but not by E2, after a 1-h incubation. The process requires the binding of apoE to the heparan sulfate proteoglycan/LDL receptor-related protein complex. The second pathway is activated after a 2-h incubation of the cells, in another isoform-dependent manner (E2 = E3 dbl greater-than sign E4) and is mediated by calcium. Our results suggest that apoE might affect cell metabolism and survival in neurons in an isoform-specific manner by inducing novel signaling pathways. PMID- 11890676 TI - Human alkaline phosphatase expression and secretion into chicken eggs after in vivo gene electroporation in the oviduct of laying hens. AB - In vivo gene electroporation was used to examine whether or not a recombinant protein is synthesized in the chicken oviduct and subsequently secreted into eggs. A plasmid DNA containing a secretion form of the human alkaline phosphatase gene was injected into mucosa of the chicken magnum. Immediately, in vivo gene electroporation was conducted. The human alkaline phosphatase activity in the oviduct mucosa increased and reached its peak at 2 days posttransfection, followed by a sharp decrease to a negligible level at 4 days posttransfection. In the egg white, the alkaline phosphatase activity showed a similar change to that in the magnum mucosa except for a delay of 4 days. The present results imply that in vivo gene electroporation method in the oviduct may serve as a rapid production system of recombinant proteins into chicken eggs. PMID- 11890677 TI - Fenton reaction is primarily involved in a mechanism of (-)-epigallocatechin-3 gallate to induce osteoclastic cell death. AB - To propose candidates for the prevention or treatment of osteoporosis, we have screened compounds naturally in food for their ability to regulate the differentiation and function of osteoclasts. One of the major green tea flavonoids, (-)-epigallocatechin-3-gallate (EGCG), was found to induce apoptotic cell death of osteoclast-like multinucleated cells after 24 h treatment in a dose dependent manner (25-100 microM), whereas osteoblasts were not affected. In the present study, we report for the first time a novel cell-death-inducing mechanism triggered by EGCG. The induction of apoptosis by EGCG was suppressed by pretreatment of catalase or calcitonin. It was also suppressed by Fe(III) and Fe(II) chelators. Furthermore, EGCG promoted the reduction of Fe(III) into Fe(II), and the combination of EGCG/Fe(III)/H(2)O(2) induced single-strand DNA breakage in a cell free system. These results indicate that the Fenton reaction is primarily involved in EGCG-induced osteoclastic cell death. PMID- 11890678 TI - Analysis of epitopes on endometrial epithelium by scanning immunoelectron microscopy. AB - Scanning immunoelectron microscopy was applied to human endometrial epithelium for the first time to simultaneously determine epitope localisation and cellular architecture. The method was established using HMFG1, an antibody to a glycoform of the MUC1 mucin. This was chosen because of the potential importance of MUC1 in connection with endometrial receptivity. Biopsies of mid-secretory phase endometrium were labelled using HMFG1 and silver-enhanced, gold-conjugated secondary antibody was then visualised by back-scattered electron imaging. The method provided a highly specific localisation of the HMFG1 epitope to the ciliated and "ciliogenic" cells of the endometrial surface. In contrast, no reactivity was evident on the microvillous cells and endometrial pinopodes. The potential to integrate the study of the molecular and ultrastructural changes that occur in the endometrium by using scanning immunoelectron microscopy offers a powerful means of expanding our understanding of the adaptation of the endometrium in preparation for embryo implantation. PMID- 11890679 TI - A novel aberrant splicing mutation of the PEX16 gene in two patients with Zellweger syndrome. AB - Human Pex16p, a peroxisomal membrane protein composed of 336 amino acids, plays a central role in peroxisomal membrane biogenesis. A nonsense mutation (R176ter) in the PEX16 gene has been reported in the case of only one patient (D-01) belonging to complementation group D of the peroxisome biogenesis disorders. We have now identified two patients belonging to group D (D-02 and D-03) whose fibroblasts were found to contain no peroxisomal membrane structure ghosts. Molecular analysis of the PEX16 gene revealed aberrant cDNA species lacking 65 bp, corresponding to exon 10 skipping caused by a splice site mutation (IVS10 + 2T - >C). Both patients, although unrelated, were homozygous for this mutation. This mutation changes the amino acid sequence starting from codon 298 and introduces a termination codon at codon 336. As a consequence, the cell's ability to membrane synthesis and protein import is disrupted, which implies that the changed C terminus of the Pex16p in these patients likely affects its function. PMID- 11890680 TI - Mouse ARF-related protein 1: genomic organization and analysis of its promoter. AB - ARF-related protein 1 (ARFRP1) is a membrane-associated GTPase which inhibits the ARF/Sec7-dependent activation of phospholipase D. We have recently shown that deletion of Arfrp1 in mice results in increased apoptosis of mesodermal cells during gastrulation, leading to early embryonic lethality. Here we describe the organization of the Arfrp1 gene and of its promoter region. The Arfrp1 gene spans approximately 7 kb and contains 8 exons. The proximal 5'-flanking regions of mouse and human ARFRP1 lack a TATA box and a CAAT box, are highly GC-rich and contain potential transcription factor binding sites. Interestingly, sequence analysis of human ARFRP1 showed its 5'-flanking region contains the first exon of another gene (DJ583P15.3 in the ensembl data base; www.ensembl.org) on the opposite strand. Promoter analysis revealed that the intergenic region between both genes (54 bp) exhibits bidirectional promoter activity. However, deletion analysis demonstrated that transcription of both genes is regulated by different cis-elements. Mutational analysis and electrophoretic mobility shift assays indicated that two short cRel- and cEts1-like elements in the 5'-flanking region of Arfrp1 (-76 to -53 and -45 to -23) are critical for the regulation of Arfrp1 expression. PMID- 11890681 TI - Clast5/Stra13 is a negative regulator of B lymphocyte activation. AB - CD40 is a member of the tumor necrosis factor receptor family and mediates a variety of functions of B cells, including B cell survival, proliferation, immunoglobulin gene class switching, memory B cell formation, and regulation of Fas-mediated apoptosis. To begin to elucidate the molecular mechanism governing such diverse functions of CD40, we have isolated a gene from mouse splenic B cells, termed Clast5, whose expression is strongly repressed during B cell activation. Clast5 is identical with Stra13, a recently identified member of the basic helix-loop-helix family of transcription factors. Clast5/Stra13 is highly expressed in unstimulated, resting B cells and is rapidly downregulated by a variety of stimuli that activate B cells, including CD40 ligand, anti-IgM antibodies, lipopolysaccharides and interleukin-4. Forced expression of Clast5/Stra13 in B cells delayed the cell cycle progression into S phase and strongly suppressed Fas-mediated apoptosis. Moreover, Clast5/Stra13 inhibited the colony formation in fibroblasts. Our results suggest that Clast5/Stra13 functions as a negative regulator of B cell activation by inhibiting cell cycle progression and cell growth. PMID- 11890682 TI - Phosphatase and oxygen radical-generating activities of mammalian purple acid phosphatase are functionally independent. AB - Bone-resorbing osteoclasts and activated macrophages express large amounts of tartrate-resistant acid phosphatase (TRAP), an iron-containing enzyme with unknown biological function. We studied acid phosphatase (AcP) and reactive oxygen species (ROS)-generating activities of recombinant rat TRAP. pH optimum was 4.5 for AcP activity and 6.5 for ROS-generating activity. Replacement of His113 and His216 by site-directed mutagenesis severely inhibited AcP activity, but had no significant effects on ROS-generating activity. Substrate specificity was not affected by the mutations. These results suggest that AcP and ROS generating activities of TRAP are functionally independent. PMID- 11890683 TI - In vitro autoradiographic localization of (125)i-secretin receptor binding sites in rat brain. AB - Although the existence of the receptor for secretin in the brain was suggested, the localization of secretin receptor and the neuronal function of secretin have not been clarified yet. In the present study, the localization of secretin receptor was investigated in the rat brain by using an in vitro autoradiography technique. Frozen section autoradiography with (125)I-secretin showed intense binding in the nucleus of solitary tract, laterodorsal thalamic nucleus, and accumbens nucleus; moderate binding in the hippocampus, caudate/putamen, cerebellum, cingulate and orbital cortices. Scatchard plot analysis gave the Kd value of 125 pM with Bmax of 134 fmol/mg tissue in the hippocampus. The binding specificity was confirmed with secretin and its analogs, VIP, PACAP, and glucagon. These results indicate the secretin receptor system might have some neural functions in the brain, which could give the basis for therapeutic use of secretin in autistic children. PMID- 11890684 TI - Transgenic expression of FGF8 and FGF10 induces transdifferentiation of pancreatic islet cells into hepatocytes and exocrine cells. AB - FGF signaling is essential for normal development of pancreatic islets. To examine the effects of overexpressed FGF8 and FGF10 on pancreatic development, we generated FGF8- and FGF10-transgenic mice (Tg mice) under the control of the glucagon promoter. In FGF8-Tg mice, hepatocyte-like cells were observed in the periphery of pancreatic islets, but areas of alpha and beta cells did not decrease, whereas in FGF10-Tg mice, pancreatic ductal and acinar cells were found in islets, concomitantly with disturbed beta-cell differentiation. These results suggest that FGF8 and FGF10 play important roles in development of hepatocytes and exocrine cells, respectively, and explain the absence of FGF8 expression in normal islets and pancreatic hypoplasia in FGF10-deficient mice. PMID- 11890685 TI - Adenoviral BMP-2 gene transfer in mesenchymal stem cells: in vitro and in vivo bone formation on biodegradable polymer scaffolds. AB - The aim of this study was to determine the feasibility of adenoviral gene transfer into primary human bone marrow osteoprogenitor cells in combination with biodegradeable scaffolds to tissue-engineer bone. Osteoprogenitors were infected with AxCAOBMP-2, a vector carrying the human BMP-2 gene. Alkaline phosphatase activity was induced in C2C12 cells following culture with conditioned media from BMP-2 expressing cells, confirming successful secretion of active BMP-2. Expression of alkaline phosphatase activity, type I collagen and mineralisation confirmed bone cell differentiation and maintenance of the osteoblast phenotype in extended culture for up to 6 weeks on PLGA porous scaffolds. In vivo implantation of adenoviral osteoprogenitor constructs on PLGA biodegradeable scaffolds, using diffusion chambers, also demonstrated bone cell differentiation and production of bone tissue. The maintenance of the osteoblast phenotype in extended culture and generation of mineralised 3-D scaffolds containing such constructs indicate the potential of such bone tissue engineering approaches in bone repair. PMID- 11890686 TI - Membrane topogenesis of the three amino-terminal transmembrane segments of glucose-6-phosphatase on endoplasmic reticulum. AB - We investigated the membrane topogenesis of glucose-6-phosphatase (G6Pase), a multispanning membrane protein, on the endoplasmic reticulum. In COS-7 cells, the first transmembrane segment (TM1) with weak hydrophobicity is inserted into the membrane in the N-terminus-out/C-terminus-cytoplasm orientation. The following TM2 is inserted depending on TM3. TM3 has the same orientation as TM1. In contrast to data from living cells, the full-length molecule and N-terminal fusion constructs were not inserted into the membrane in a cell-free system. Addition of a signal recognition particle did not improve G6Pase insertion. When the 37-residue N-terminal segment was deleted, however, TM2 and TM3 were correctly inserted. We concluded that the three N-terminal TM segments are inserted into the membrane dependent on the two signal-anchor sequences of TM1 and TM3. TM1 is likely to be an unconventional signal sequence that barely functions in vitro. The 37-residue N-terminal segment inhibits the signal function of the following TM3 in cell-free systems. PMID- 11890687 TI - Gelatinase levels in male and female breast cancer. AB - Breast cancer is a common disease in females but very rare in males, in whom it shows a more metastatic behavior, and a worse prognosis. Matrix metalloprotease-2 (MMP-2) and MMP-9 are proteolytic enzymes balanced by tissue inhibitor of MMP-2 (TIMP-2), commonly involved in cancer metastasis. This is the first study on gelatinolytic activity in male breast cancer patients, compared to that in female patients. In cancer tissues, both gelatinases were more expressed than in normal samples, being and more concentrated in male than in female patients. TIMP-2 levels were slightly increased in normal compared to those in cancer tissues and more concentrated in males than in females. Immunostaining showed that in male cancer tissues MMP-2 and MMP-9 staining was more intense and diffuse than in female cancer tissues, while no differences were observed regarding TIMP-2. In conclusion, the increased expression of gelatinases in male breast cancer patients together with anatomical features might explain the high tendency toward metastasis and the worse prognosis. PMID- 11890688 TI - Extracellular pH modulates Helicobacter pylori-induced vacuolation and VacA toxin internalization in human gastric epithelial cells. AB - In this study we investigated whether an acidic extracellular pH may inhibit H. pylori-induced internalization of bacterial virulence factors by gastric epithelium, thus preventing ingestion of potentially dangerous luminal contents and resulting cellular damage. The interaction of H. pylori VacA toxin and ammonia (produced by H. pylori urease) with partly polarized gastric MKN 28 cells in culture was investigated at neutral and moderately acidic pH (6.2, compatible with cell viability) by means of neutral red dye uptake and ultrastructural immunocytochemistry. We found that acidic extracellular pH virtually abolished both VacA-dependent and ammonia-dependent cell vacuolation, as shown by the neutral red test, and caused a 50% decrease in VacA internalization into endosomal vesicles and vacuoles, as assessed by quantitation of immunogold particles. In addition, acidic pH blocked endosomal internalization of H. pylori outer membrane vesicles, a convenient indicator of endocytosis. Our data raise the possibility that suppression of gastric acid may increase H. pylori-induced gastric damage by enhancing epithelial internalization of H. pylori virulence factors through activation of endocytosis. Increased transmembrane diffusion of ammonia could also contribute to this process. PMID- 11890689 TI - Smoking has no effect on the amino acid composition of apolipoprotein B100 of LDL while directly influencing the antioxidant status. AB - Previous studies have demonstrated increased plasma levels of oxidised low density lipoprotein (oxLDL) in chronic smokers, which has been associated with the extent of endothelial dysfunction. In this study we examine the relationship between the amino acid composition of apolipoprotein B100 (apo B) of low-density lipoprotein (LDL), by reverse phase HPLC after precolumn derivatisation, between smokers (> or =40 cigarettes/day) and nonsmokers in relation to their plasma and LDL antioxidant status. While there was a significant difference in the levels of plasma vitamin C and alpha-tocopherol between female smokers and nonsmokers, as well as in the levels of LDL alpha-tocopherol, there was no significant difference in the amino acid composition of apo B between the two groups. PMID- 11890690 TI - IL-2-mediated upregulation of uPA and uPAR in natural killer cells. AB - Urokinase plasminogen activator (uPA) and its receptor uPAR play a major role in immune cell-mediated, including natural killer (NK) cell-mediated, degradation of extracellular matrices. Herein, we investigate the effects of IL-2 on NK cell uPA and uPAR. RNA and protein analyses showed upregulation of uPA and uPAR following IL-2 stimulation. Gel-shift assays and Western blots detected uPA and uPAR mRNA binding proteins (mRNABPs), previously shown to destabilize uPA and uPAR mRNA. Following IL-2 stimulation, a downregulation of uPAR mRNABP and a reciprocal induction of uPAR mRNA were noted. The increase in uPA following IL-2 stimulation appeared to be more transcriptionally regulated. These data suggest that IL-2 upregulates both uPA and uPAR in NK cells through posttranscriptional as well as transcriptional mechanisms, partially explaining increases in NK cell invasiveness following IL-2 stimulation. PMID- 11890691 TI - Digoxin up-regulates MDR1 in human colon carcinoma Caco-2 cells. AB - Because MDR1 (P-glycoprotein) plays an important role in pharmacokinetics such as absorption and excretion of xenobiotics and multidrug resistance, an understanding of the factors regulating its function and expression is important. Here, the effects of digoxin on cell sensitivity to an anticancer drug, MDR1 function, and expression were examined by assessing the growth inhibition by paclitaxel, the transport characteristics of the MDR1 substrate Rhodamine123, and the level of MDR1 mRNA, respectively, using human colon carcinoma Caco-2 cells, which are widely used as a model of intestinal epithelial cells. The sensitivity to paclitaxel, an MDR1 substrate, in Caco-2 cells pretreated with digoxin was lower than that in non-treated cells. The accumulation of Rhodamine123 was reduced by pretreatment with digoxin and its efflux was enhanced. The level of MDR1 mRNA in Caco-2 cells was increased in a digoxin concentration-dependent manner. These results taken together suggested that digoxin up-regulates MDR1 in Caco-2 cells. PMID- 11890692 TI - Subcellular localization of beta-catenin is regulated by cell density. AB - It is generally accepted that subcellular distribution of beta-catenin regulates its function. Membrane-bound beta-catenin mediates cell-cell adhesion, whereas elevation of the cytoplasmic and nuclear pool of the protein is associated with an oncogenic function. Although the role of beta-catenin in transformed cells is relatively well characterized, little is known about its importance in proliferation and cell-cycle control of nontransformed epithelial cells. Using different approaches we show that in human keratinocytes (HaCaT) beta-catenin is distributed throughout the cells in subconfluent, proliferating cultures. In contrast, beta-catenin is nearly exclusively located at the plasma membrane in confluent, contact-inhibited cells. Hence, we demonstrate for the first time that beta-catenin is translocated from the cytoplasm to the plasma membrane in response to high cell density. We conclude that beta-catenin plays an important role in proliferation and mediating contact-inhibition by changing intracellular localization. PMID- 11890694 TI - Tefluthrin modulates a novel anionic background conductance (I(AB)) in guinea-pig ventricular myocytes. AB - This report describes for the first time a novel anionic background current (I(AB)) identified in guinea-pig isolated ventricular myocytes. It also shows that I(AB) has both novel and differential pharmacology from other (cardiac) chloride currents. Using the whole-cell patch-clamp technique and external anion substitution, I(AB) was found to be outwardly rectifying and highly permeable to NO(-)(3), with a relative permeability sequence of NO(-)(3) > I(-) > Cl(-). I(AB) was not blocked by 50 microM DIDS, by hypertonic external solution, or by the nonselective protein kinase inhibitor H7-DHC. Exposure to the pyrethroid agent tefluthrin (10 microM) increased the current density of I(AB) significantly at positive voltages (P < 0.05), but had no significant effect on other cardiac chloride currents. We conclude that I(AB) possesses a distinct pharmacology and does not fall into the three major classes of cardiac chloride conductance commonly reported. PMID- 11890693 TI - Activator protein-1 complex expressed by magnetism in cultured rat hippocampal neurons. AB - Brief exposure for 15 min to static magnetic filed at 100 mT led to marked but transient potentiation of binding of a radiolabeled probe for activator protein-1 (AP1) in immature cultured rat hippocampal neurons with high expression of growth associated protein-43. Immunoblotting and supershift analyses revealed that brief exposure to static magnetic field increased AP1 DNA binding through expression of Fra-2, c-Jun, and Jun-D proteins in immature cultured hippocampal neurons. Significantly less potent increases were seen in both intracellular free Ca(2+) concentration and AP1 binding following the addition of N-methyl-d-aspartate in these immature neurons exposed to magnetism 24 h before. These results suggest that brief exposure to weak static magnetic field may lead to desensitization of NMDA receptor channels through modulation of de novo synthesis of particular inducible target proteins at the level of gene transcription by the AP1 complex expressed in the nucleus of immature cultured rat hippocampal neurons. PMID- 11890695 TI - Pulmonary inflammation monitored noninvasively by MRI in freely breathing rats. AB - A detailed analysis has been carried out of the correlation between the signals detected by MRI in the rat lung after allergen or endotoxin challenge and parameters of inflammation determined in the broncho-alveolar lavage (BAL) fluid. MRI signals after allergen correlated highly significantly with the BAL fluid eosinophil number, eosinophil peroxidase activity and protein concentration. Similar highly significant correlations were seen when the anti-inflammatory glucocorticosteroid, budesonide, manifested against allergen. In contrast, following endotoxin challenge, mucus was the sole BAL fluid parameter that correlated significantly with the long lasting signal detected by MRI. Since edema is an integral component of pulmonary inflammation, MRI provides a noninvasive means of monitoring the course of the inflammatory response and should prove invaluable in profiling anti-inflammatory drugs in vivo. Further, the prospect of noninvasively detecting a sustained mucus hypersecretory phenotype in the lung brings an important new perspective to models of chronic obstructive pulmonary diseases. PMID- 11890696 TI - Minimal heparin/heparan sulfate sequences for binding to fibroblast growth factor 1. AB - The glycosaminoglycans heparin and heparan sulfate (HS) bind to fibroblast growth factor FGF1 and promote its dimerization, a proposed prerequisite for binding to a cellular receptor and triggering mitogenic signals. The problem of minimal structural requirements for heparin/HS sequences to bind FGF1 was approached by surface plasmon resonance (SPR), NMR spectroscopy, and MALDI mass spectrometry studies using the three synthetic tetrasaccharides GlcNSO(3)6OR-IdoA2SO(3) GlcNSO(3)6OR'-IdoA2SO(3)OPr (AA, R = R' = SO(3); BA, R = H, R' = SO(3); BB, R = R' = H; Pr, propyl). AA and BA significantly interact with the protein, whereas BB is practically inactive. The NMR spectra show that, whereas the interaction of AA primarily involves the GlcNSO(3)6SO(3)IdoA2SO(3) disaccharide moiety at its nonreducing end, residues at both the nonreducing (NR) and reducing side (R) appear to be involved in the weaker complex of BA. Furthermore, MALDI experiments show that, in addition to 1:1 protein:tetrasaccharide complexes, AA and BA are able to form 2:1 complexes, indicating that heparin/HS-induced dimerization of FGF1 requires only one 6-OSO(3) group per tetrasaccharide. PMID- 11890697 TI - Novel, not adenylyl cyclase-coupled cannabinoid binding site in cerebellum of mice. AB - In this study we report data suggesting the presence of a non-CB1, non-CB2 cannabinoid site in the cerebellum of CB1-/- mice. We have carried out [(35)S]GTPgammaS binding experiments in striata, hippocampi, and cerebella of CB1 /- and CB1(+/+) mice with Delta(9)-THC, WIN55,212-2, HU-210, SR141716A, and SR144528. In CB1-/- mice Delta(9)-THC and HU-210 did not stimulate [(35)S]GTPgammaS binding. However, WIN55,212-2 was able to stimulate [(35)S]GTPgammaS binding in cerebella of CB1-/- mice. The maximal effect of this stimulation was 31% that of wild type animals. This effect was reversible neither by CB1 nor CB2 receptor antagonists. Similar results were obtained with the endogenous cannabinoid, anandamide. However, adenylyl cyclase was not inhibited by WIN55,212-2 or anandamide in the CB1(minus sign/minus sign) animals. In striata and hippocampi of CB1-/- mice no [(35)S]GTPgammaS stimulation curve could be obtained with WIN55,212. Our findings suggest that there is a non-CB1 non-CB2 receptor present in the cerebellum of CB1-/- mice. PMID- 11890698 TI - Temporal generation of multiple antifungal proteins in primed seeds. AB - A drastic increase of antifungal activity was demonstrated during plant seed germination and in seed protein extract in vitro. Multiple antifungal proteins with a wide spectrum of activity were generated and identified. Chromatographic and electrophoretic analysis demonstrated that during seed germination, more fractions with potent antifungal activity were generated, and the antifungal activity shifted from small molecules to high molecular proteins. This germination-related increase of antifungal activity were observed in all three plants tested, i.e., cheeseweed, cigar tree and wheat. This rapid increase of antifungal activity was also observed with incubation of seed proteins in vitro, suggesting that at least part of the antifungal protein generation is independent of gene expression. Seven antifungal proteins with activities against five different plant pathogens were isolated from the active fractions. However, random digestion of purified seed protein with multiple proteinases failed to generate any antifungal proteins. It is suggested that during plant seed germination, a regulated biochemical process takes place that results in the generation of multiple peptides or proteins with antifungal activities. This onset of antifungal proteins is transitional in nature, but could play an important role in the protection of plants in early stage of development when the more sophisticated defense system has yet to develop. PMID- 11890699 TI - Multiple liver-specific factors bind to a 64-bp element and activate apo(a) gene. AB - The high plasma levels of lipoprotein(a) [Lp(a)] are associated with atherosclerosis. The apo(a) gene is responsible for the variance of Lp(a) concentration and its expression is liver-specific. By 5'-deletion analysis, we, in a luciferase gene reporter assay, have identified a 64-bp AT-rich region of upstream apo(a) gene (-703 to -640) that binds to multiple liver-specific factors. The 64 bp cis-element contained three dyad symmetry elements (DSEs) that are crucial for synergistic binding to the factors. We have demonstrated that both DSE-2 and -3 together are responsible for factor binding in vitro, and for gene activation in liver cells. Further, we have purified one of the UV cross linked DNA-protein complexes to homogeneity by streptavidin magnetic bead chromatography. The identification of a further upstream negative regulatory region (-1432 to -704) led us to predict that as yet unidentified transcriptional repressor(s) might also repress apo(a) gene transcription. PMID- 11890700 TI - Localization of a negative vitamin D response sequence in the human growth hormone gene. AB - 1,25-Dihydroxyvitamin D(3) [1,25(OH)(2) D(3)] exerts its biological effects by binding to the vitamin D receptor (VDR), which binds in turn to the vitamin D response elements located in the target gene's promoter. We have previously demonstrated that VDR binds in vitro with high affinity to the 5'-flanking sequence of the human growth hormone (hGH) gene. In this study, we analyzed the response to 1,25(OH)(2) D(3) of hGH-promoter constructs introduced by transfection into the MCF-7 human adenocarcinoma cell line. We found that the transcriptional activity of some of these constructs was markedly reduced by 1,25(OH)(2) D(3). Deletion analyses revealed that a 34-bp sequence located between positions -62 and -29 upstream of the transcription start site is sufficient for this repressive response. This conclusion was also confirmed by gel mobility shift assays. Our results indicate that vitamin D inhibits hGH gene transcription, directly or by interference with other transcription factors. PMID- 11890701 TI - Murine Apg12p has a substrate preference for murine Apg7p over three Apg8p homologs. AB - Apg7p is a unique E1 enzyme which is essential for both the Apg12p- and Apg8p modification systems, and plays indispensable roles in yeast autophagy. A cDNA encoding murine Apg7p homolog (mApg7p) was isolated from a mouse brain cDNA library. The predicted amino acid sequence of the clone shows a significant homology to human Apg7p and yeast Apg7p. Murine Apg12p as well as the three mammalian Apg8p homologs co-immunoprecipitate with mApg7p. Site-directed mutagenesis revealed that an active-site cysteine within mApg7p is Cys(567), indicating that mApg7p is an authentic E1 enzyme for murine Apg12p and mammalian Apg8p homologs. The mutagenesis study also revealed that Apg12p has a substrate preference for mApg7p over the three Apg8p homologs, suggesting that the Apg12p conjugation by Apg7p occurs preferentially in mammalian cells compared with the modification of the three Apg8p homologs. We also report here on the ubiquitous expression of human APG7 mRNA in human adult and fetal tissues and of rat Apg7p in adult tissues. PMID- 11890702 TI - An experimental validation of orphan genes of Buchnera, a symbiont of aphids. AB - Although Buchnera sp. APS, an intracellular symbiont of pea aphids, is a close relative of Escherichia coli, its genome has been extensively modified because of its prolonged intracellular life. In our previous studies on the Buchnera genome, computer analysis predicted three "orphan" genes, yba2, yba3, and yba4, which are open reading frames (ORFs) with no homologs in the database. In this paper, we successfully validated all these orphan genes by RT-PCR and Northern hybridization. The present study also revealed that yba3 and yba4 formed an operon, suggesting that they function in concert. Sequences around transcriptional start sites suggests that these genes are under the control of sigma 70. In view of codon usage and AT bias observed in these genes, it is likely that Buchnera have maintained them for an evolutionarily long time. PMID- 11890703 TI - Nucleosome linker proteins HMGB1 and histone H1 differentially enhance DNA ligation reactions. AB - We previously reported that HMGB1, which originally binds to chromatin in a manner competitive with linker histone H1 to modulate chromatin structure, enhances both intra-molecular and inter-molecular ligations. In this paper, we found that histone H1 differentially enhances ligation reaction of DNA double strand breaks (DSB). Histone H1 stimulated exclusively inter-molecular ligation reaction of DSB with DNA ligase IIIbeta and IV, whereas HMGB1 enhanced mainly intra-molecular ligation reaction. Electron microscopy of direct DNA-protein interaction without chemical cross-linking visualized that HMGB1 bends and loops linear DNA to form compact DNA structure and that histone H1 is capable of assembling DNA in tandem arrangement with occasional branches. These results suggest that differences in the enhancement of DNA ligation reaction are due to those in alteration of DNA configuration induced by these two linker proteins. HMGB1 and histone H1 may function in non-homologous end-joining of DSB repair and V(D)J recombination in different manners. PMID- 11890704 TI - Retinoic acid-inducible gene-I is induced in endothelial cells by LPS and regulates expression of COX-2. AB - Bacterial lipopolysaccharides (LPS) induce expression of multiple genes in endothelial cells, which are critical cellular effectors in various pathologic syndromes. Using subtractive hybridization to identify genes that are differentially induced in human endothelial cells treated with LPS, we found that retinoic acid-inducible gene I (RIG-I) is induced in endothelial cells stimulated with LPS. RIG-I encodes a protein belonging to the DExH-box family which has diverse roles in regulation of gene expression and cellular functions. Cyclooxygenase-2 (COX-2) is also induced in endothelial cells by LPS. Overexpression of RIG-I selectively upregulated expression of COX-2 and also induced COX-2 promoter activity. RIG-I is an inducible gene in stimulated endothelial cells that may have important roles in vascular pathology by virtue of its ability to regulate expression of the COX-2 gene product. PMID- 11890705 TI - A novel five-subunit-type 2-oxoglutalate:ferredoxin oxidoreductases from Hydrogenobacter thermophilus TK-6. AB - A thermophilic, chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, fixes carbon dioxide via the reductive TCA cycle. 2-Oxoglutarate:ferredoxin oxidoreductase (OGOR) of this strain is one of the key enzymes of the pathway. OGOR of strain TK-6 has been reported to be a two subunit-type OGOR and encoded by korAB. A gene cluster, forDABGEF, encoding another OGOR was found 148 bp upstream of korAB in the opposite orientation. Five of the for genes (forDABGE) were required for the expression of the active recombinant enzyme in Escherichia coli. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed five polypeptides corresponding to the forDABGE gene products, suggesting that the enzyme had a novel five-subunit structure. The recombinant enzyme had high substrate specificity toward 2 oxoglutarate as in the case of the gene products of korAB. Primer extension analysis showed that the korA and forD genes were transcribed from one and two transcriptional initiation sites, respectively. The results also suggested that both gene clusters were expressed in the cells of strain TK-6. PMID- 11890706 TI - IL-6 induces expression of the Fli-1 proto-oncogene via STAT3. AB - Induction of gene expression by IL-6 has become an area of intense interest due to the role this cytokine plays in mediating aspects of inflammation, cellular differentiation, and proliferation. The ETS family of transcription factors represents a group of positive and negative regulators of transcription, that are differentially expressed in a cell and tissue specific manner. The ETS protein Fli-1 is known to induce differentiation in the erythroblastic leukemia cell line K562 along megakaryocytic developmental pathways. Here we show that IL-6 treatment of K562 induces the expression of Fli-1 via the STAT3 transcription factor. Upregulation of Fli-1 expression can be abrogated by the addition of AG490, a chemical inhibitor of JAK kinases, and by transfecting the cells with a dominant negative STAT3 expression construct. PMID- 11890707 TI - Animal model of sclerotic skin induced by bleomycin: a clue to the pathogenesis of and therapy for scleroderma? PMID- 11890708 TI - Millennium award recipient contribution. Identification of children with early onset and high incidence of anti-islet autoantibodies. AB - A total of 21,000 general population newborns (NECs) and 693 young siblings offspring of patients with type 1A diabetes (SOCs) were class II genotyped and 293 NECs and 72 SOCs with the high-risk genotype, DR3/4, DQB1*0302 have been prospectively evaluated. Seventeen individuals who converted to persistent autoantibody positivity and two autoantibody-negative control groups (35 SOCs and 24 NECs) were typed for HLA-A class I alleles. The A1, A2 genotype was significantly increased among the autoantibody-positive subjects (47%) compared to autoantibody-negative SOCs (14%, P = 0.01) and NECs (13%, P = 0.02). Life table analysis of DR3/4, DQB1*0302 siblings revealed a risk of 75% for development of islet autoantibodies by the age of 2 years for those with A1, A2. The HLA-A2 phenotype frequency was increased among an independent DR3/4, DQB1*0302 young diabetes cohort (64% versus 33% for autoantibody-negative NECs). These results suggest that a high incidence and early appearance of islet autoantibodies for siblings of patients with type 1A diabetes are associated with DR3/4, DQB1*0302 and potentially increased with HLA-A genotype A1, A2. PMID- 11890709 TI - Thalidomide induces granuloma differentiation in sarcoid skin lesions associated with disease improvement. AB - Sarcoidosis, a chronic granulomatous disease of unknown etiology, is treated with immune suppressive drugs such as corticosteroids. Sarcoidosis patients have been reported to benefit clinically from treatment with thalidomide. We administered thalidomide for 16 weeks to eight patients with chronic skin sarcoidosis and evaluated the drug's effects before and with treatment. After thalidomide treatment, all skin biopsies showed decreases in granuloma size and reduction in epidermal thickness. We also observed extensive T cell recruitment into the granulomas, the appearance of multinucleated giant cells, and increased numbers of dermal Langerhans cells (CD1a(+)) and mature dendritic cells (CD83(+) or DC LAMP(+)). Plasma IL-12 levels increased and remained elevated during the treatment period. We noted increased HLA-DR expression on peripheral blood lymphocytes and a corresponding drop in the naive T cell marker CD45RA. Our data suggest that thalidomide treatment of sarcoidosis results in granuloma differentiation to a Th1-type cellular immune response usually associated with protective immunity to tuberculosis and tuberculoid leprosy. PMID- 11890710 TI - Colitis-related public T cells are selected in the colonic lamina propria of IL 10-deficient mice. AB - IL-10 is an important regulatory cytokine in the mucosal immune system, as supported by the fact that mice deficient in IL-10 spontaneously develop Crohn's disease-like colitis. An aberrant, Th1-driven CD4(+) T-cell response to enteric bacteria seems to be important in the pathogenesis of this murine colitis. However, no specific bacteria or bacterial products have been identified, and whether the colitis is mediated by the activation of CD4(+) T cells that recognize specific peptide-MHC complexes is controversial. In this study, we analyzed the TCR beta chain complementarity determining region 3 length spectratype of colonic CD4(+) T cells isolated from diseased IL-10-deficient mice by using the Immunoscope technique. Screening of the diseased interleukin-10 deficient mice resulted in a restricted clonotype in TCR V beta 13 and 14 subfamilies of colonic CD4(+) T cells. In contrast, a Gaussian distribution of clonotype of individual TCR V beta subsets was observed in CD4(+) T cells from the peripheral lymphoid tissues. Although individual variability in the disease related response was also noted in other IL-10-deficient mice maintained in La Jolla and Osaka, perhaps because of different stages of the disease, genetic background, or the housing environment, colitis-related public clones seemed to be shared in all the diseased mice tested. To address whether public clones were involved, we determined the DNA sequence of the clones. Public motifs were shared in colonic CD4(+) T cells from different background interleukin-10-deficient mice with colitis. The frequently found motifs were SXDWG and SATGNYAEQ. These motifs were not seen in the peripheral lymphoid tissues of diseased mice as well as the colon of non-diseased mice. Thus, the common motif may be related to a public gut derived antigen, which could be important for the development of pathogenic CD4(+) T cells in this inflammatory bowel disease (IBD) model. The selection of V beta-J beta usage is perhaps stochastic in individual mice; however, the epigenetic generation of SXDWG motif by the recombination machinery and selection for this motif in the gut environment could be important for triggering IBD. PMID- 11890711 TI - Kinetics of B, CD4 T, and CD8 T cells infused into humans: estimates of intravascular:extravascular ratios and total body counts. AB - Little is known about the rate of T and B cell traffic from blood to extravascular compartments or about the steady-state distribution of T and B cells between intravascular and extravascular compartments in humans. We quantitated circulating T and B cell subsets before and during the first 24 h after the infusion of an allogeneic or syngeneic peripheral blood stem cell graft (containing approximately 10(10) lymphocytes) into 10 patients conditioned with chemotherapy and/or total body irradiation. For all lymphocyte subsets measured, <15% of the infused cells were present in the blood at the end of the 0.5-h infusion and <3% of the infused cells were present in the blood 1 h later. Thereafter, CD4 T cell counts plateaued at approximately 1% and CD8 T cell counts at < or = 0.4% of the infused cells, whereas B cell counts declined slowly (1.5% of the infused B cells were present in the blood at 2 h and 0.3% at 24 h postinfusion). We conclude that the rate of lymphocyte traffic from blood to extravascular spaces can be extraordinary (approximately 10(10) lymphocytes can leave blood within 0.5 h) and that at steady state the blood contains approximately 1% total body CD4 T cells, < or = 0.4% total body CD8 T cells, and approximately 1.4% total body B cells. By inference, an average-size person may carry a total of approximately 4.1 x 10(11) CD4 T cells, > or = 4.5 x 10(11) CD8 T cells, and approximately 1.0 x 10(11) B cells. PMID- 11890712 TI - Attenuated production of intracellular IL-10 and IL-12 in monocytes from patients with severe asthma. AB - Interleukin (IL)-10 and IL-12 production is decreased in peripheral blood mononuclear cells of patients with mild asthma. Using whole blood culture and flow cytometry we examined whether monocyte heterogeneity influenced IL-10 and IL 12 production in subjects with severe asthma. We demonstrated that IL-10 release in LPS-stimulated whole blood culture was decreased in patients with severe persistent asthma compared to those with mild asthma and controls (P = 0.04 and P < 0.001, respectively). In asthmatic patients, the percentage of CD14(+)CD16(+) cells was higher than that from normal subjects (P = 0.04). Severe asthmatics showed significantly less positive staining for IL-10 and IL-12 (P < 0.001 and P = 0.02, respectively) after stimulation in monocytes, compared to mild asthmatics and controls in both CD14(+)CD16(+) and CD14(+)CD16(minus sign) cells. These results suggest that IL-10 synthesis is attenuated in severe persistent asthma compared to mild asthma and that this cannot be explained by the increase in the CD14(+)CD16(+) monocytes in asthma. PMID- 11890713 TI - A non-class I MHC intestinal epithelial surface glycoprotein, gp180, binds to CD8. AB - The activation of CD8(+) T cells by normal intestinal epithelial cells in antigen specific or allogeneic mixed cell culture systems has significant implications for the modulation of mucosal immune responses due to the fact that these T cells appear to have regulatory rather than cytolytic activity. A 180-kDa glycoprotein (gp180) has been identified and shown to be important in CD8(+) T cell activation by intestinal epithelial cells. In this study, we examine, in further detail, the role that the CD8 molecule plays in this interaction. It has been previously shown that monoclonal antibodies against gp180 inhibited the activation of CD8 associated p56(lck) in T cells. Although indirectly suggested by these data, there was no evidence that the activation of this protein tyrosine kinase was a direct result of gp180 interacting with the CD8 molecule. In this study, we document that soluble gp180 is able to bind to CD8-Fc fusion proteins and is absorbed by human CD8 alpha but not CD4 transfected murine T cells and that this interaction is dependent upon carbohydrate on the gp180 molecule. Furthermore, the sites used for binding by gp180 are distinct from those used by the conventional CD8 ligand, class I MHC. Thus, gp180 appears to be a novel CD8 ligand that plays an important role in the activation of CD8-associated kinases and of CD8(+) T cells. PMID- 11890714 TI - Estrogen inhibits systemic T cell expression of TNF-alpha and recruitment of TNF alpha(+) T cells and macrophages into the CNS of mice developing experimental encephalomyelitis. AB - Estrogen treatment has been found to have suppressive activity in several models of autoimmunity. To investigate the mechanism of 17 beta-estradiol (E2) suppression of experimental autoimmune encephalomyelitis, we evaluated E2 effects on TNF-alpha expression in the central nervous system (CNS) and spleen of C57BL/6 mice immunized with MOG 35-55/CFA. Kinetic analysis demonstrated that E2 treatment drastically decreased the recruitment of total inflammatory cells as well as TNF-alpha(+) macrophages and T cells into the CNS at disease onset. In contrast, E2 had only moderate effects on the relatively high constitutive TNF alpha expression by resident CNS microglial cells. E2 treatment also had profound inhibitory effects on expression of TNF-alpha by splenic CD4(+) T cells, including those responsive to MOG 35-55 peptide. We propose that the mechanism of E2 protection may involve both systemic inhibition of TNF-alpha expression and local (CNS) recruitment of inflammatory cells, with modest effects on TNF-alpha expression by resident CNS microglial cells. PMID- 11890715 TI - Expression of cytokine- and chemokine-related genes in peripheral blood mononuclear cells from lupus patients by cDNA array. AB - Systemic lupus erythematosus (SLE) is characterized by diverse and complex immune abnormalities. In an effort to begin to characterize the full complexity of immune abnormalities, the expression pattern of 375 potentially relevant genes was analyzed using peripheral blood mononuclear cells (PBMC) from 21 SLE patients and 12 controls by cDNA arrays. When mean gene expression for patients was compared to controls, 50 genes were identified that exhibited more than 2.5-fold difference in expression level. By the Mann-Whitney U test, 20 genes were significantly different (P < 0.05) between patients and controls. Most of these genes have not been previously associated with SLE and belong to a variety of families such as TNF/death receptor, IL-1 cytokine family, and IL-8 and its receptors. Hierarchical clustering of samples and differentially expressed genes revealed that with few exceptions, patients clustered separately from controls. These results highlight the potential use of the microarray data in identifying genes associated with SLE, which could become candidate molecular markers or future therapeutic targets. PMID- 11890716 TI - Tumor-associated macrophages as a source of functional dendritic cells in ovarian cancer patients. AB - We have generated dendritic cells (DCs) from tumor-associated macrophages (TAMs) obtained from ascites fluid or tumor specimens of ovarian cancer patients, and compared them phenotypically and functionally to DCs derived from the patients' peripheral blood mononuclear cells (PBMCs). Both immature and mature DCs could be generated from TAMs. However, TAM-derived DCs underwent maturation to a lesser degree than PBMC-derived DCs, as measured by DC receptor surface expression. Nonetheless, in allogeneic mixed lymphocyte reactions, TAM-derived DCs stimulated T cell proliferation as efficiently as PBMC-derived DCs. In addition, TAM-derived DCs presenting tumor Ags were capable of stimulating IFN-gamma secretion by tumor specific T cell lines. Thus, TAMs isolated from ovarian cancer patients can be used to generate significant numbers of DCs. Therefore, TAMs have potential use for either ex vivo or in vivo DC-based immunotherapy, particularly in individuals in which more conventional sources of DCs have been depleted. PMID- 11890718 TI - Autoimmune responses in patients with linear IgA bullous dermatosis: both autoantibodies and T lymphocytes recognize the NC16A domain of the BP180 molecule. AB - Linear IgA bullous disease (LABD) is an autoimmune skin disease characterized by subepidermal blisters and IgA autoantibodies directed against the epidermal basement membrane zone (BMZ) of the skin. Various antigens have been identified as targets of IgA autoantibodies including BP180, a type II glycoprotein that spans the BMZ and lamina lucida. Previously, we have identified a subset of LABD patients whose sera contained IgA antibodies against the 16th noncollagenous (NC16A) domain of BP180. NC16A was previously shown to harbor epitopes that are recognized by both autoantibodies and T cells from patients with bullous pemphigoid and herpes gestationis and is thought to be associated with the development of these immunobullous diseases. The aim of this study was to determine whether T lymphocytes from LABD patients with anti-NC16A IgA autoantibodies respond to epitopes in the same region of the BP180 protein. Indeed, of the four LABD patients in our study, all had T cells that specifically proliferated in response to NC16A. Moreover, two subfragments of NC16A were identified as the predominant targets of LABD T cells. Further analysis of T cell lines and clones derived from these patients revealed that these cells express a CD4 memory T cell phenotype and secrete a Th1/Th2 mixed-cytokine profile, characteristics similar to those of T cells in bullous pemphigoid patients. Our data suggest that the BP180 protein, typically the NC16A region, is the common target of both cellular and humoral immune responses in some LABD patients. This information helps to further elucidate the autoimmune mechanisms in this disease. PMID- 11890717 TI - Impaired T cell function in RANTES-deficient mice. AB - The chemokine RANTES is a chemoattractant for monocytes and T cells and is postulated to participate in many aspects of the immune response. To evaluate the biological roles of RANTES in vivo, we generated RANTES-deficient (-/-) mice and characterized their T cell function. In cutaneous delayed-type hypersensitivity assays, a 50% reduction in ear and footpad swelling was seen in -/- mice compared to +/+ mice. In vitro, polyclonal and antigen-specific T cell proliferation was decreased. Quantitative analysis using the fluorescent dye carboxy-fluorescein succinimidyl ester revealed that this proliferative defect was due both to fewer antigen-reactive T cells and to a reduction in the capacity of these cells to proliferate. In addition, IFN-gamma and IL-2 production by the -/- T cells was dramatically decreased. Together, these data suggest that RANTES is required for normal T cell functions as well as for recruiting monocytes and T cells to sites of inflammation. PMID- 11890719 TI - Quantitative analysis of adhesion molecules on cellular constituents of the human uterine microenvironment under the influence of estrogen and progesterone. AB - The uterus contains all the components of a tertiary lymphoid compartment. We hypothesize that specific leukocyte recruitment to the endometrium during the secretory phase of the menstrual cycle and early pregnancy limits the type of immunocyte that gains access. The present study utilized flow cytometry to define and quantify adhesion molecules possibly used by decidual infiltrating lymphocytes (DIL) as homing receptors, uterine microvascular myometrial endothelial cells (UtMVE-Myo) as addressins, and secretory endometrial stroma cells (STO) as retainment factors. Human umbilical cord vein endothelial cells and peripheral blood lymphocytes were used as control cells for comparison studies. DIL were composed of predominantly lymphocyte function-associated antigen (LFA)-1+, intercellular adhesion molecule (ICAM)-1+, LFA-2+, LFA-3+, gp150,95+, alpha1beta1+, Hermes cell adhesion molecule (H-CAM)+, and neural cell adhesion molecule (N-CAM)+ (CD56(bright)) memory/effector natural killer cells. A significant number of UtMVEC-Myo expressed platelet endothelial cell adhesion molecule (PECAM)-1, a percentage were uniquely LFA-3+, and alpha4 integrin expression was uniquely high. An increased number of STO uniquely expressed alpha3, beta3, and LFA-3, whereas alpha2, alpha4, alphaVbeta3, and H-CAM were significantly increased. Possible unique adhesions of DIL:UtMVEC-Myo included SLe(x):PECAM, vascular cell adhesion molecule-1:alpha4, and LFA-2:LFA-3, whereas DIL:STO included LFA-2:LFA-3 and N-CAM:N-CAM. Unique molecules on DIL may also associate with extracellular matrix (ECM) or complement on UtMVEC-Myo or STO to form gp150,95:fibrinogen/iC3b/C3dg, alpha1beta1:laminin (LM)/collagen (CO), and ICAM-1:fibronectin (FN) interactions. Bridges of ECM may also form between DIL and UtMVEC-Myo adhesion molecules including ICAM-1:FN:ICAM-1 and alpha4beta1:FN:alpha4beta1. DIL:ECM:STO interactions may involve alpha2beta1:CO:alpha2beta1, alpha3beta1:LM/CO/FN:alpha3beta1, alphaVbeta3:VN:alphaVbeta3, and H-CAM:hyaluronate:H-CAM. It is likely that many adhesion molecules play a role in the recruitment and retainment of specialized lymphocytes within the uterine microenvironment. (Mackay et al., 1990). PMID- 11890720 TI - Gene expression of tropoelastin is enhanced in the aorta proximal to the coarctation in rabbits. AB - To assess elastin biosynthesis in the aortic wall in response to acute elevation of blood pressure, we studied the aortic gene expression of tropoelastin in a rabbit midthoracic aortic coarctation model. The time points of the study were 1, 3, and 7 days and 2, 4, and 8 weeks after coarctation. Additional animals were subjected to hypercholesterolemia for analysis of tropoelastin expression in the intimal lesion. mRNA for tropoelastin was quantitated by Northern blot analysis and its distribution was revealed by in situ hybridization. The 65-kDa tropoelastin was analyzed by Western blotting and immunohistochemistry. Tropoelastin mRNA proximal to the coarctation was increased at 2 weeks and returned to baseline by 8 weeks (P < 0.05 versus control). Changes in 65-kDa tropoelastin corresponded to those of mRNA. Tropoelastin gene was expressed mainly in the intima and in the outer media at the proximal region to the stenoses, which was particularly remarkable in the intimal lesion. The results indicate that tropoelastin gene expression was enhanced in the early remodeling response to elevated blood pressure. The distribution of newly synthesized tropoelastin in the outer media suggests a reenforcement role of tropoelastin, which preserves mechanical resiliency in response to changes in tensile stress. PMID- 11890722 TI - Parasitic adaptive mechanisms in infection by leishmania. AB - Leishmania are a resilient group of intracellular parasites that infect macrophages. The resultant complex of diseases, or leishmaniases, caused by the parasites affect over twelve million people worldwide. Leishmania have developed unique adaptive mechanisms to ensure their survival in the harsh environments faced throughout their life cycle. These parasites must not only contend with the hostile digestive conditions found within the sand fly vector, but they must also avoid destruction by the host immune system while in the bloodstream, before entering the macrophage. To do so, Leishmania express unique lipophosphoglycan (LPG) molecules and the metalloprotease gp63, among other proteins, on their cell surface. To enter the macrophage, Leishmania utilizes a variety of cellular receptors to mediate endocytosis. Once inside the macrophage, Leishmania is protected from phagolysosome degradation by a variety of adaptations to inhibit cellular defense mechanisms. These include the inhibition of phagosome-endosome fusion, hydrolytic enzymes, cell signaling pathways, nitric oxide production, and cytokine production. While other parasites can also infect macrophages, Leishmania is distinctive in that it not only relies on its own defenses to survive and reproduce within the macrophage phagolysosome, but Leishmania also manipulates the host immune response in order to protect itself and to gain entry into the cell. These unique adaptive mechanisms help promote Leishmania survival. PMID- 11890721 TI - Cellular and molecular alterations in spinal cord injury patients with pressure ulcers: a preliminary report. AB - The study was designed to investigate the changes, both numerically and functionally, of the molecules critical to wound healing in spinal cord injury (SCI) patients. Spinal cord injury patients who demonstrated delayed healing of their pressure ulcers were used as study subjects. Age-matched healthy individuals served as controls. Adhesion molecule expression of the peripheral blood leukocytes, including lymphocytes and granulocytes, was measured by flow cytometric analysis. Binding capacity of the lymphocytes was evaluated using human umbilical cord vein endothelial cells (HUVECs) as the binding matrix. Samples from pressure ulcers of the patients were immunostained to define fibronectin, kalinin, beta4 integrin, alpha2beta1, alpha3beta1, alpha5beta1, and CD138 expression. Compared to healthy controls, there was decreased expression of CD11a, CD11b, CD18, CD49b, CD49c, CD49d, CD54, and CD8 in patients' lymphocyte populations and CD11a, CD18, CD49c, CD49d, and CD8 in patients' granulocyte populations. The binding capacity, expressed as percentage binding of the lymphocytes to the HUVEC matrix, was greatly diminished in the patients. There was markedly diminished immunohistochemical staining of fibronectin in pressure ulcers. These findings showed that delayed healing of pressure ulcers in SCI patients can be attributed to reduced adhesion molecule expression, impaired cell cell interaction, and lack of extracellular matrix structural and functional protein. PMID- 11890723 TI - Expression of lumican in thickened intima and smooth muscle cells in human coronary atherosclerosis. AB - Lumican is a member of a small leucine-rich proteoglycan family. Members of this family play an important role in cell migration and proliferation during embryonic development, tissue repair, and tumor growth. Lumican is reported to be overexpressed during the wound-healing process in the cornea and ischemic and reperfused heart. Recently, we found that lumican mRNA and its protein are expressed in cultured vascular smooth muscle cells (VSMCs) from the rat aorta. However, the expression and role of lumican in human atherosclerotic tissues are not clearly elucidated. In the present study, we aimed to clarify whether lumican is expressed in VSMCs and its localization in human coronary atherosclerotic tissues. The lumican protein and its mRNA were expressed in a small number of VSMCs in the media of normal coronary artery, but the lumican protein was not localized in the medial stroma. In contrast, the lumican protein and its mRNA were expressed in most of VSMCs that migrated into the thickened intima, but not in infiltrating foamy macrophages. The lumican protein was prominently localized in the thickened intimal stroma. The lumican protein and its mRNA were also expressed in VSMCs in the inner layer of the media and its protein was localized in medial stromal tissues. These findings indicate that the lumican protein is mainly synthesized by intimal and medial VSMCs in coronary atherosclerosis and that lumican contributes to collagen fibrillogenesis of coronary atherosclerosis. PMID- 11890724 TI - Subnormal shear stress-induced intimal thickening requires medial smooth muscle cell proliferation and migration. AB - Arterial intimal thickening is consisted of predominately smooth muscle cells (SMC). The source of these SMCs and mechanisms response for their changes have not been well cleared. Using a model of rabbit common carotid artery (CCA) shear induced intimal thickening, we sought to identify and describe the source of SMCs in intima. The enlarged CCA 28 days after arteriovenous fistula (AVF) creation was subjected to subnormal wall shear stress (WSS) for 1, 3, and 7 days by closure of the AVF. To determine SMC proliferation, BrdU pulse labeling of SMCs was performed. BrdU-labeled SMCs were tracked over time to further confirm SMC migration. In response to subnormal WSS intimal thickening developed progressively. BrdU-labeled SMCs localized in the subendothelial area. When the BrdU-labeled medial SMCs were tracked 1 day after AVF closure, progenies of these BrdU-incorporated SMCs increased by 4.8-fold with 75% of them in the intima. They were 12-fold increased with 83% in the intima 7 days after. En face examination showed an accumulation of SMCs in internal elastic lamina gap after AVF closure, which later migrated into subendothelial area. In situ hybridization revealed increased TGF-beta1 mRNA expression in intimal SMCs. This study demonstrates that the medial SMCs are the predominant cells in subnormal WSS-induced intimal thickening. Early expression of TGF-beta1 may play an important role in the process of intimal thickening. PMID- 11890725 TI - Distribution of silicon/e in tissues of mice of different fibrinogen genotypes following intraperitoneal administration of emulsified poly(dimethylsiloxane) [correction of poly(dimethysiloxane)]. AB - Following injection into the abdominal cavity of a C57BL/6 mouse, droplets of emulsified PDMS visible by light microscopy (diameter > or = 1 microm) disseminate to multiple organs of the animal. Because fibrinogen may facilitate dissemination, we compared histologically the accumulation of PDMS droplets in lymph nodes, lungs, spleen, liver, and left kidney of Fib +/+, Fib +/-, and Fib /- mice of C57BL/6 background 35 and 75 days after intraperitoneal injection of an emulsion of the polymer. We also used ICP-AES to assess the accumulation of silicon in the lymph nodes, livers, and spleens of the animals. The emulsion droplets ranged in diameter from approximately 0.04 to approximately 80 microm. PDMS droplets visible by light microscopy were in all organs of both Fib +/+ mice and Fib +/- mice. In those animals, droplets were invariably either within or adjacent to inflammatory cells, predominantly macrophages. In contrast, PDMS droplets were visible in none of the organs of Fib -/- mice. Despite the absence of visible droplets in them, the lymph nodes, livers, and spleens of Fib -/- mice, like the corresponding organs of Fib +/+ and Fib +/- mice, contained measurable silicon after 35 and 75 days. The amount of silicon, however, was always greater in the organs of Fib +/+ and Fib +/- mice than in the organs of Fib -/- mice. We attribute the presence of silicon in organs that had no histologic evidence of droplets to diffusion of the very smallest droplets/soluble species of PDMS from the abdominal cavity. Taken together, our data and observations implicate a role for fibrinogen in the dissemination of larger PDMS droplets in vivo. We propose this role involves recognition of droplet-bound fibrinogen by macrophages and, perhaps, other inflammatory cells, and the subsequent fibrinogen-facilitated ingestion and/or extracellular movement of the droplets by those cells. PMID- 11890727 TI - On balance. PMID- 11890726 TI - Gastrointestinal stromal tumors: are they of cajal cell origin? AB - Recently some reports have suggested that gastrointestinal stromal tumors (GIST) might originate from the interstitial cells of Cajal or differentiate into them because they express c-kit and/or CD34 and indicated that the majority of previously diagnosed smooth muscle tumors (SMT) actually belong to GIST, but are not true SMT. We, therefore, detected c-kit, CD34, SMA, and S-100 in 106 Chinese cases of gastrointestinal tumors, which were histopathologically diagnosed as smooth muscle tumors originally, to demonstrate the immunophenotypes of these tumors. The results showed that 73 cases had immunoreaction with c-kit and/or CD34, of which 48 cases showed coexpression with either SMA or S-100 or with both. A correlation between the immunophenotypes and known histopathological parameters was also shown here based on follow-up data. We suggest that the concept of GIST should not be used as an umbrella to cover all gastrointestinal mesenchymal tumors, but be defined in a narrow term as differing from true smooth muscle tumors. PMID- 11890728 TI - The development of children's rule use on the balance scale task. AB - Cognitive development can be characterized by a sequence of increasingly complex rules or strategies for solving problems. Our work focuses on the development of children's proportional reasoning, assessed by the balance scale task using Siegler's (1976, 1981) rule assessment methodology. We studied whether children use rules, whether children of different ages use qualitatively different rules, and whether rules are used consistently. Nonverbal balance scale problems were administered to 805 participants between 5 and 19 years of age. Latent class analyses indicate that children use rules, that children of different ages use different rules, and that both consistent and inconsistent use of rules occurs. A model for the development of reasoning about the balance scale task is proposed. The model is a restricted form of the overlapping waves model (Siegler, 1996) and predicts both discontinuous and gradual transitions between rules. PMID- 11890729 TI - Young children's performance on the balance scale: the influence of relational complexity. AB - Three experiments investigated the effect of complexity on children's understanding of a beam balance. In nonconflict problems, weights or distances varied, while the other was held constant. In conflict items, both weight and distance varied, and items were of three kinds: weight dominant, distance dominant, or balance (in which neither was dominant). In Experiment 1, 2-year-old children succeeded on nonconflict-weight and nonconflict-distance problems. This result was replicated in Experiment 2, but performance on conflict items did not exceed chance. In Experiment 3, 3- and 4-year-olds succeeded on all except conflict balance problems, while 5- and 6-year-olds succeeded on all problem types. The results were interpreted in terms of relational complexity theory. Children aged 2 to 4 years succeeded on problems that entailed binary relations, but 5- and 6-year-olds also succeeded on problems that entailed ternary relations. Ternary relations tasks from other domains--transitivity and class inclusion--accounted for 93% of the age-related variance in balance scale scores. PMID- 11890730 TI - Development of rules and strategies: balancing the old and the new. AB - The experiments described in the lead articles replicate findings from previous studies of development of knowledge about balance scales, add several new findings, and raise four key questions: (a) How can rule use best be assessed? (b) How can we reconcile systematic use of rules with variable use of strategies? (c) When do children begin to use rules? and (d) How do children generate new rules? In this Reflection, we summarize current understanding of development of knowledge about balance scales and consider each of the four questions. PMID- 11890731 TI - The balance beam in the balance: reflections on rules, relational complexity, and developmental processes. AB - The lead articles by Jansen and van der Maas (2002, this issue) and Halford, Andrews, Dalton, Boag, and Zielinski (2002, this issue) raise numerous questions concerning the development of rule use and how best to assess it. In this Reflection, we focus on the following: (a) When can one infer the use of a rule? (b) What are the mechanisms underlying the development of rule use? and (c) What is the relation between understanding and execution? In addressing these questions, we contrast relational complexity theory with cognitive complexity and control (CCC) theory. PMID- 11890732 TI - Bridging the gap between theory and model: a reflection on the balance scale task. AB - At first glance, the two lead articles in this issue share little except the balance scale task, yet we view them as complementary rather than unrelated or contradictory. Our Reflection focuses on the individual strengths of the two lead articles and, to a greater extent, the potential power of their combined perspectives. Our general approach is to allow psychological theory to suggest a model of performance that can both evaluate specific theoretical claims and reveal important features of the data that had been previously obscured using conventional statistical analyses. Our guiding principle is that model, theory, and data all should be connected. PMID- 11890733 TI - Counting in sign language. AB - Do the visuomanual modality and the structure of the sequence of numbers in sign language have an impact on the development of counting and its use by deaf children? The sequence of number signs in Belgian French Sign Language follows a base-5 rule while the number sequence in oral French follows a base-10 rule. The accuracy and use of sequence number string were investigated in hearing children varying in age from 3 years 4 months to 5 years 8 months and in deaf children varying in age from 4 years to 6 years 2 months. Three tasks were used: abstract counting, object counting, and creation of sets of a given cardinality. Deaf children exhibited age-related lags in their knowledge of the number sequence; they made different errors from those of hearing children, reflecting the rule bound nature of sign language. Remarkably, their performance in object counting and creating sets of given cardinality was similar to that of hearing children who had a longer sequence number string, indicating a better use of counting than predicted by their knowledge of the linguistic sequence of numbers. PMID- 11890734 TI - Does the nature of the experience influence suggestibility? A study of children's event memory. AB - Two experiments examined the effects of event modality on children's memory and suggestibility. In Experiment 1, 3- and 5-year-old children directly participated in, observed, or listened to a narrative about an event. In an interview immediately after the event, free recall was followed by misleading or leading questions and, in turn, test recall questions. One week later children were reinterviewed. In Experiment 2, 4-year-old children either participated in or listened to a story about an event, either a single time or to a criterion level of learning. Misleading questions were presented either immediately or 1 week after learning, followed by test recall questions. Five-year-old children were more accurate than 3-year-olds and those participating were more accurate than those either observing or listening to a narrative. However, method of assessment, level of event learning, delay to testing, and variables relating to the misled items also influenced the magnitude of misinformation effects. PMID- 11890735 TI - Mechanisms of dysfunction of the nitric oxide pathway in vascular diseases. AB - Vascular nitric oxide (NO) is involved in many physiologic and pathophysiologic processes throughout the body. Many vascular diseases have a reduction in the activity of endothelium-derived NO as an important component involved in the initiation and/or progression of the disease. It is now known that there are multiple mechanisms for this reduction in NO activity with one or more mechanisms operating depending on the specific condition or stage of a disease. In other instances, the therapy for certain diseases is responsible for the reduction in NO activity and contributes to the acceleration of vascular disease. This review details the known mechanisms of dysfunction of the NO pathway leading to vascular diseases, which provides the rationale for why certain therapies can improve while other therapies adversely affect vascular health. PMID- 11890736 TI - 3-Morpholinosydnonimine hydrochloride induces p53-dependent apoptosis in murine primary neural cells: a critical role for p21(ras)-MAPK-p19(ARF) pathway. AB - In some neurological disorders, excessive nitric oxide (NO, nitrogen monoxide) produced by inducible and/or neuronal nitric oxide synthases (iNOS and nNOS) is able to combine with superoxide (O(minus sign)(2)) to form peroxynitrite (ONOO(minus sign)), which can then induce p53-dependent neural apoptosis. In the present study, experiments using p53 knock-out mice primary neural cells revealed that 3-morpholinosydnonimine hydrochloride (SIN-1), a peroxynitrite donor, triggered apoptosis, while p53-transcriptional activity was effectively suppressed in the absence of p53 molecules. This shows that SIN-1 was able to induce p53-dependent apoptosis in murine primary neural cells. The mechanism responsible for the SIN-1-induced accumulation of p53 molecules was then analyzed. Western blot analysis indicated that p53 accumulation caused by SIN-1 did not require p53 phosphorylation, whereas SIN-1 treatment triggered MAP kinase (MAPK) phosphorylation and pretreatment with the MAP kinase kinase (MEK) inhibitor U0126 inhibited p53 accumulation. Pretreatment of the neural cells with lovastatin, an inhibitor of p21(ras) signaling, greatly inhibited the accumulation of p53 induced by SIN-1. Northern blot and immunofluorescence analyses revealed that primary neural cells treated with SIN-1 had increased levels of p19 alternate reading frame (p19(ARF)) mRNA and protein, which is induced by MAPK and stabilizes the p53 protein. Our findings clearly show that the p21(ras)-MAPK-p19(ARF) pathway has an essential role in p53-dependent apoptosis triggered by peroxynitrite in neural cells. PMID- 11890737 TI - Release of nitric oxide from novel diazeniumdiolates monitored by laser magnetic resonance spectroscopy. AB - We describe a technique in which for the first time laser magnetic resonance spectroscopy (LMRS) is used online to monitor the release of nitric oxide from synthetic NO donors. LMRS is a spectroscopic method for selective, sensitive (to 1 ppb), and time-resolved NO gas detection in the far-infrared and midinfrared spectral regions. We used two partly novel sets of diazeniumdiolates, the first set derived from N,N-diethylamine (-->DEA-NO), piperidine ((-->PIPE-NO), 2 methylpiperidine (-->MEPIPE-NO), and 2-ethylpiperidine (-->EPIPE-NO) and the second set derived from 2-, 3-, and 4-piperidine carboxylic acids (-->PIPECO-NO, NIPECO-NO, ISONIPECO-NO). We monitored the acid-catalyzed NO liberation from these compounds as influenced by parameters such as pH, temperature, concentration, and molecular structure. PIPECO-NO turned out to be the fastest donor of the group. The 3- and 4-substituted isomer derivatives were only negligibly faster releasers than unsubstituted PIPE-NO, which on the other hand showed to be faster than the higher homologues MEPIPE-NO and EPIPE-NO. The results demonstrate that varying neighboring groups affect the functional diazeniumdiolate group differently. A vicinal carboxyl group increases and alkyl groups decrease the rate of NO release. PMID- 11890738 TI - Heparin-binding EGF-like growth factor down regulates proinflammatory cytokine induced nitric oxide and inducible nitric oxide synthase production in intestinal epithelial cells. AB - We have previously shown that heparin-binding EGF-like growth factor (HB-EGF) protects intestinal epithelial cells (IEC) from necrosis and apoptosis in vitro and from intestinal ischemia/reperfusion injury in vivo; however, the mechanisms of HB-EGF cytoprotection are unclear. Overproduction of iNOS and NO have been implicated in the pathogenesis of several forms of ischemia/reperfusion injury. We therefore studied whether HB-EGF could down-regulate proinflammatory cytokine induced iNOS and NO production in intestinal epithelial cells in vitro. DLD-1 human intestinal epithelial cells were exposed to the proinflammatory cytokines interleukin-1beta (IL-1beta) (20 ng/ml) and interferon-gamma (IFN-gamma) (10 ng/ml) to stimulate iNOS induction and NO production. Cells were treated with HB EGF (0-100 ng/ml) either before or with cytokine exposure, and cells and supernatants were harvested 24 and 48 h later. Accumulated NO was measured in supernatants by chemiluminescence. Total RNA was extracted from cell lysates for iNOS mRNA quantification using real-time reverse transcription-polymerase chain reaction (RT-PCR), and total protein was extracted from cell lysates for detection of iNOS protein. HB-EGF significantly decreased cytokine-induced NO production in a dose dependent manner, and NO reduction was associated with iNOS suppression at both the mRNA and protein levels. While cytokine exposure resulted in a significant increase in iNOS mRNA expression in these cells (109 plus minus 9 fold), HB-EGF reduced iNOS expression by 5.7-fold (P < 0.05). These results suggest that HB-EGF may exert its cytoprotective effects, in part, by down regulating iNOS and NO production, and provides further rationale for additional testing of the effects of HB-EGF in the treatment of intestinal ischemia/reperfusion injury in vivo. PMID- 11890739 TI - Nitric oxide gas decreases endothelin-1 mRNA in cultured pulmonary artery endothelial cells. AB - Inhaled nitric oxide gas (iNO) vasodilates the pulmonary circulation. The effective "dose" of iNO for chronic treatment of pulmonary hypertension is unknown. Increased abundance of pulmonary mRNA for preproendothelin-1 (ppET-1) with its associated increase in endothelin-1 (ET-1) could contribute to the development of both clinical and experimental pulmonary hypertension. The benefit of iNO therapy may be from inhibition of ET-1 production. The present study was designed to compare the effects of two therapeutic concentrations of NO gas, 10 parts per million (p.p.m.) and 100 p.p.m. on the steady-state level of mRNA for ppET-1 and nitric oxide synthase (NOS III), in cultured bovine pulmonary artery endothelial cells. Uptake of NO gas was assessed by measurement of nitrite anions in the medium. The mRNA for ppET-1 and NOS III was determined by semiquantitative reverse-transcriptase polymerase chain reaction (RT-PCR). After 4 h exposure to 100 p.p.m. NO in air, nitrite anions levels were 1.6 microM in the endothelial cell media as opposed to 0.23 microM with 10 p.p.m. NO. The levels were 0.02 microM in control cells exposed to air alone. Exposure to 100 p.p.m. NO reduced the steady state levels of mRNA for ppET-1, but not NOSIII mRNA levels. By comparison 10 p.p.m. NO did not affect levels of either mRNA. PMID- 11890740 TI - Effect of inhibitors of nitric oxide synthase on acetaminophen-induced hepatotoxicity in mice. AB - We recently reported that following a toxic dose of acetaminophen to mice, tyrosine nitration occurs in the protein of cells that become necrotic. Nitration of tyrosine is by peroxynitrite, a species formed from nitric oxide (NO) and superoxide. In this manuscript we studied the effects of the NO synthase inhibitors N-monomethyl-l-arginine (l-NMMA), N-nitro-l-arginine methyl ester (NAME), l-N-(1-iminoethyl)lysine (l-NIL), and aminoguanidine on acetaminophen hepatotoxicity. Acetaminophen (300 mg/kg) increased serum nitrate/nitrite and alanine aminotransferase (ALT) levels, indicating increased NO synthesis and liver necrosis, respectively. None of the NO synthase inhibitors reduced serum ALT levels. In fact, l-NMMA, l-NIL, and aminoguanidine significantly augmented acetaminophen hepatotoxicity at 4 h. A detailed time course indicated that aminoguanidine (15 mg/kg at 0 h and 15 mg/kg at 2 h) significantly increased serum ALT levels over that for acetaminophen alone at 2 and 4 h; however, at 6 and 8 h serum ALT levels in the two groups were identical. At 2 h following acetaminophen plus aminoguanidine NO synthesis was significantly increased; however, at 4, 6, and 8 h NO synthesis was significantly decreased. Aminoguanidine also decreased acetaminophen-induced nitration of tyrosine. Acetaminophen alone did not induce lipid peroxidation, but acetaminophen plus aminoguanidine significantly increased hepatic lipid peroxidation (malondialdehyde levels) at 2, 4, and 6 h. These data are consistent with NO having a critical role in controlling superoxide-mediated lipid peroxidation in acetaminophen hepatotoxicity. Thus, acetaminophen hepatotoxicity may be mediated by either lipid peroxidation or by peroxynitrite. PMID- 11890742 TI - Anticoagulants and other preanalytical factors interfere in plasma nitrate/nitrite quantification by the Griess method. AB - Nitric oxide (NO) is a signal molecule with functions such as neurotransmission, local vascular relaxation, and anti-inflammation in many physiological and pathological processes. Various factors regulate its intracellular lifetime. Due to its high reactivity in biological systems, it is transformed in the bloodstream into nitrates (NO(-)(3)) by oxyhemoglobin. The Griess reaction is a technically simple method (spectrophotometric, 540 nm) for the analysis of nitrites (NO(-)(2)) in aqueous solutions. We studied the interference of common anticoagulants in the quantification of nitrate and nitrite in plasma samples by the Griess method. We obtained rat plasma using heparin or sodium EDTA as anticoagulants, then added, or otherwise, known NO(-)(3) amounts in order to calculate their recovery. We also studied the effect of ultra-filtration performed before Griess reaction on plasma and aqueous solutions of various anticoagulants (heparin, EDTA, and also sodium citrate) to compare the recoveries of added NO(-)(3) or NO(-)(2). We used standards of NO(-)(3) or NO(-)(2) for quantification. We conclude that: (i) The bacterial nitrate reductase used to reduce NO(-)(3) to NO(-)(2) is unstable in certain storage conditions and interferes with different volumes of plasma used. (ii) The ultrafiltration (which is sometimes performed before the Griess reaction) of plasma obtained with EDTA or citrate is not recommended because it leads to overestimation of NO(minus sign)(3). In contrast, ultrafiltration is necessary when heparin is used. (iii) The absorbance at 540 nm attributed to plasma itself (basal value or background) interferes in final quantification, especially when ultrafiltration is not performed. For the quantification of plasma NO(-)(3) we recommend: sodium EDTA as anticoagulant, no ultrafiltration of plasma, and measurement of the absorbance background of each sample. PMID- 11890741 TI - Regulation of eNOS in normal and diabetes-impaired skin repair: implications for tissue regeneration. AB - An important role of inducible nitric oxide (NO) synthase for epithelial action during skin repair has been well established. Although a delayed healing of skin wounds has been recently described for eNOS-deficient mice, a participation of endothelial-type NO synthase (eNOS) in skin repair largely remains unclear. In this study we determined the expression pattern of eNOS during wound healing in healthy and in diabetic mice. Remarkably, normal repair in healthy animals was characterized by a moderate induction of eNOS at the mRNA and protein level, whereas diabetes-impaired healing was associated with a clearly reduced eNOS protein expression. Immunohistochemistry revealed the endothelial lining of blood vessels within the granulation tissue, and also keratinocytes of the wound margins, the developing neo-epithelium, and the hair follicles to express eNOS protein. Keratinocyte-derived expression of eNOS could be confirmed at the mRNA level in vitro for human primary keratinocytes and the keratinocyte cell line HaCaT. Furthermore, eNOS enzymatic activity most likely contributes to epithelial regeneration, as eNOS-deficient (eNOS -/-) animals exhibited reduced wound margin epithelia associated with reduced keratinocyte proliferation. PMID- 11890743 TI - Protein thiols and glutathione influence the nitric oxide-dependent regulation of the red blood cell metabolism. AB - Nitric oxide (NO) can modulate red blood cell (RBC) glycolysis by translocation of the enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPD) (EC 1.2.1.12) from the cytosolic domain of the membrane protein band 3 (cdb3) in the cytosol. In this study we have investigated which NO-reactive thiols might be influencing GAPD translocation and the specific role of glutathione. Two highly reactive Cys residues were identified by transnitrosylation with nitrosoglutathione (GSNO) of cdb3 and GAPD (K(2) = 73.7 and 101.5 M(-1) s(-1), respectively). The Cys 149 located in the catalytic site of GAPD is exclusively involved in the GSNO-induced nitrosylation. Reassociation experiments carried out at equilibrium with preparations of RBC membranes and GAPD revealed that different NO donors may form -SNO on, and decrease the affinity between, GAPD and cdb3. In intact RBC, the NO donors 3-morpholinosydnonimine (SIN-1) and peroxynitrite (ONOO(-)) significantly increased GAPD activity in the cytosol, glycolysis measured as lactate production, and energy charge levels. Our data suggest that ONOO(-) is the main NO derivative able to cross the RBC membrane, leading to GAPD translocation and SNO formation. In cell-free experiments and intact RBC, diamide (a thiol oxidant able to inhibit GAPD activity) was observed to reverse the effect of SIN-1 on GAPD translocation. The results demonstrate that cdb3 and GAPD contain reactive thiols that can be transnitrosylated mainly by means of GSNO; these can ultimately influence GAPD translocation/activity and the glycolytic flux. PMID- 11890744 TI - Role of nitric oxide in control of light adaptive cone photomechanical movements in retinas of lower vertebrates: a comparative species study. AB - The possible role of nitric oxide (NO) as a novel light adaptive neuromodulator of cone plasticity (photomechanical movements) in retinae of two contrasting species of fish (freshwater carp and marine bream) and an example of an amphibian (Xenopus laevis) was studied pharmacologically by cytomorphometric measurements. Application of a NO donor [S-nitroso-N-acetyl-d, l-penicillamine] (500-700 microM) to dark-adapted retinae induced contraction of cones with an efficiency (CE) relative to full light adaptation of around 54% in all three species. Pretreatment with a NO scavenger [2-(4-Carboxyphenyl)-4,4,5,5 tetrametylimidazoline-1-oxil-3-oxide] (30-35 microM) produced a consistent significant inhibition of the light adaptation-induced cone contraction (CE = 15 20%). These results strongly suggest the involvement of endogenous NO in the cone contractions that occur in freshwater and marine fish and amphibian retinae as a part of the light adaptation process. PMID- 11890745 TI - Hydrogen peroxide induces a rapid production of nitric oxide in mung bean (Phaseolus aureus). AB - Nitric oxide (NO) has recently been identified as an important signaling molecule in plant immune response. The present study aims to investigate the signaling pathway that leads to NO production. Using the NO specific fluorescent dye DAF 2DA, we observed rapid production of NO in mung bean leaves after the addition of 10 mM hydrogen peroxide (H(2)O(2)). NO was probably produced by a NOS-like enzyme in plants, as the NO production was inhibited by l-NAME, a NOS inhibitor. The NOS like activity in the total leaf protein preparation of mung bean (Phaseolus aureus) was elevated 8.3-fold after 10 mM H(2)O(2) treatment, as demonstrated using the chemiluminescence NOS assay. The NOS-like activity was BH(4) dependent: omitting BH(4) in the reaction mixture of NOS assay reduced the NOS activity by 76%. We also found that the H(2)O(2) induced NO production was mediated via calcium ion flux, as it was blocked in the presence of a calcium ion channel blocker, verapamil. Results from the present study identified H(2)O(2) as an upstream signal that leads to NO production in plants. H(2)O(2) and NO, besides acting as two independent signaling molecules in plant immune response, may interrelate to form an oxidative cell death (OCD) cycle. PMID- 11890746 TI - Streptozotocin-pancreatic damage in the rat: modulatory effect of 15-deoxy delta12,14-prostaglandin j(2) on nitridergic and prostanoid pathway. AB - 15-deoxy-delta (12,14)prostaglandin J(2) (15d-PGJ(2)) has been identified as a natural ligand of the PPARgamma subtype. PPAR activation in nonadipose tissues seems to inhibit iNOS and COX2 expression. Vasoactive compounds like nitric oxide and prostaglandins are increased in pancreatic tissue from streptozotocin diabetic rats. We hypothesize that 15d-PGJ(2) may regulate the production of these proinflammatory compounds that lead to beta cell destruction in the diabetic pathology. In this work we evaluated Ca(2+)-dependent (cNOS) and Ca(2+) independent (iNOS) activity, nitrate/nitrite levels, 15-dPGJ(2) and prostaglandin E(2) (PGE(2)) levels in isolated pancreatic islets, and 15d-PGJ(2) levels in plasma from control and streptozotocin-diabetic rats. Our results show that cNOS is predominant in control, while iNOS isoform is increased in the diabetic islets (P < 0.01). 15d-PGJ(2) 10(-5)M inhibits cNOS and iNOS activity both in control and diabetic islets (P < 0.05). Nitrate/nitrite and PGE(2) levels are higher in diabetic than in control islets (P < 0.05 and P < 0.01, respectively). 15d-PGJ(2) 10(-5)M decreases nitrate/nitrite and PGE(2) levels both in control and in diabetic islets. Bisphenol A diglycidyl ether (BADGE), a recently described PPARgamma antagonist, seems to act as a PPARgamma agonist, diminishing nitrate/nitrite and PGE2 levels in control and diabetic islets. 15d-PGJ(2) production is lower in islets from diabetic animals compared to control (P < 0.05). Our observations suggest that 15d-PGJ(2) is able to diminish the production of vasoactive proinflammatory agents in pancreatic islets. The diminished 15d-PGJ(2) levels in the diabetic islets are probably related to the diminished capacity to limit the inflammatory response due to experimental diabetes in the rat. PMID- 11890747 TI - The nitration product 5-nitro-gamma-tocopherol is increased in the Alzheimer brain. AB - Oxidative stress and quasi-inflammatory processes recently have been recognized as contributing factors in the pathogenesis of Alzheimer's disease (AD). Reactive nitrating species have specifically been implicated in AD based on immunochemical and instrumental detection of nitrotyrosine in AD brain protein. The significance of lipid-phase nitration has not been investigated in AD. This study documents a significant two- to threefold increase in the lipid nitration product 5-nitro gamma-tocopherol in affected regions of the AD brain as determined by high performance liquid chromatography with electrochemical detection. In a bioassay to compare the relative potency of alpha-tocopherol and gamma-tocopherol against nitrative stress, rat brain mitochondria were exposed to the peroxynitrite generating compound SIN-1. The oxidation-sensitive Kreb's cycle enzyme alpha ketoglutarate dehydrogenase was inactivated by SIN-1, in a manner that could be significantly attenuated by gamma-tocopherol (at <10 microM) but not by alpha tocopherol. These data indicate that nitric oxide-derived species are significant contributors to lipid oxidation in the AD brain. The findings are discussed in reference to the neuroinflammatory hypothesis of AD and the possible role of gamma-tocopherol as a major lipid-phase scavenger of reactive nitrogen species. PMID- 11890748 TI - Kinetics studies of the reaction of the ruthenium porphyrin Ru(OEP)(CO) with the S-nitrosothiol N-acetyl-1-amino-2-methylpropyl-2-thionitrite. AB - The reaction of the S-nitrosothiol compound N-acetyl-1-amino-2-methylpropyl-2 thionitrite (RSNO) with the model metalloporphyrin complex Ru(II)(OEP)(CO) (OEP = octaethylporphyrinato dianion) gives the addition product trans Ru(II)(OEP)(NO)(SR). Here we report the details of a stopped flow kinetics investigation which demonstrates the rapid equilibrium formation of an intermediate concluded to be S-bound RSNO complex Ru(II)(OEP)(RSNO)(CO), which undergoes a rate-limiting step, presumably S-NO bond cleavage to give a second intermediate Ru(III)(OEP)(SR)(CO) too short lived for direct observation. Notably, this is different from the nitrogen coordination pathway often proposed and represents an alternative mechanism by which S-nitrosothiols may be formed or decomposed in the presence of redox active metal centers. Also reported is a brief study of the quantitative photochemistry of RSNO, the photodecomposition of which complicates the kinetics studies by spectroscopic techniques. PMID- 11890749 TI - Nasal nitric oxide levels in cystic fibrosis patients are associated with a neuronal NO synthase (NOS1) gene polymorphism. AB - Nitric oxide (NO) plays an important role in a number of physiological processes in the airways, including host defense. Although the exact cellular and molecular source of the NO formation in airways is unknown, there is recent evidence that neuronal NO synthase (NOS1) contributes significantly to NO in the lower airways of cystic fibrosis (CF) patients. NOS1 protein has been shown to be expressed in nasal epithelium, suggesting an involvement of NOS1-derived NO in upper airway biology. We here hypothesized that nasal NO concentrations in CF patients are related to genotype variants in the NOS1 gene. Measurements of nasal NO concentration and pulmonary function were performed in 40 clinically stable CF patients. Genomic DNA from all patients was screened for an intronic AAT-repeat polymorphism in the NOS1 gene using polymerase chain reaction and simple sequence length polymorphism (SSLP) analysis. The allele size at that locus was significantly (P = 0.001) associated with upper airway NO. Mean (+/- SD) nasal NO concentrations were 40.5 +/- 5.2 ppb in CF patients (n = 12) with high repeat numbers (i.e., both alleles > or =12 repeats) and 72.6 +/- 7.4 ppb in patients (n = 28) with low repeat numbers (i.e., at least one allele <12 repeats). Furthermore, in the group of CF patients harboring NOS1 genotypes associated with low nasal NO, colonization of airways with P. aeruginosa was significantly more frequent than in patients with NOS1 genotypes associated high nasal NO concentrations (P = 0.0022). We conclude that (1) the variability in CF nasal NO levels are related to naturally occurring variants in the NOS1 gene, and (2) that nasal NOS1-derived NO affects the susceptibility of CF airways to infection with P. aeruginosa. PMID- 11890750 TI - Nitric oxide-scavenging properties of some chalcone derivatives. AB - The implication of NO in many inflammatory diseases has been well documented. We have previously reported that some chalcone derivatives can control the iNOS pathway in inflammatory processes. In the present study, we have assessed the NO scavenging capacity of three chalcone derivatives (CH8, CH11, and CH12) in a competitive assay with HbO(2), a well-known physiologically relevant NO scavenger. Our data identify these chalcones as new NO scavengers. The estimated second-order rate constants (k(s)) for the reaction of the three derivatives with NO is in the same range as the value obtained for HbO(2), with CH11 exerting the greatest effect. These results suggest an additional action of these compounds on NO regulation. PMID- 11890751 TI - Electron spin catalysis. PMID- 11890752 TI - Kinetics of cross-polarization in solid-state NMR: a guide for chemists. PMID- 11890753 TI - Reactions of alpha-diketones and omicron-quinones with phosphorus compounds. PMID- 11890754 TI - Ordered anion adlayers on metal electrode surfaces. PMID- 11890756 TI - Green fluorescent protein (GFP): applications, structure, and related photophysical behavior. PMID- 11890758 TI - The behavior of 1,n-enynes in the presence of transition metals. PMID- 11890755 TI - Supramolecular materials via polymerization of mesophases of hydrated amphiphiles. PMID- 11890757 TI - Chem-bioinformatics: comparative QSAR at the interface between chemistry and biology. PMID- 11890759 TI - Subphthalocyanines: singular nonplanar aromatic compounds-synthesis, reactivity, and physical properties. PMID- 11890761 TI - Membrane protein microarrays. AB - This paper describes the fabrication of microarrays consisting of G protein coupled receptors (GPCRs) on surfaces coated with gamma-aminopropylsilane (GAPS). Microspots of model membranes on GAPS-coated surfaces were observed to have several desired properties-high mechanical stability, long range lateral fluidity, and a thickness corresponding to a lipid bilayer in the bulk of the microspot. GPCR arrays were obtained by printing membrane preparations containing GPCRs using a quill-pin printer. To demonstrate specific binding of ligands, arrays presenting neurotensin (NTR1), adrenergic (beta1), and dopamine (D1) receptors were treated with fluorescently labeled neurotensin (BT-NT). Fluorescence images revealed binding only to microspots corresponding to the neurotensin receptor; this specificity was further demonstrated by the inhibition of binding in the presence of excess unlabeled neurotensin. The ability of GPCR arrays to enable selectivity studies between the different subtypes of a receptor was examined by printing arrays consisting of three subtypes of the adrenergic receptor: beta1, beta2, and alpha2A. When treated with fluorescently labeled CGP 12177, a cognate antagonist analogue specific to beta-adrenergic receptors, binding was only observed to microspots of the beta1 and beta2 receptors. Furthermore, binding of labeled CGP 12177 was inhibited when the arrays were incubated with solutions also containing ICI 118551, and in a manner consistent with the higher affinity of ICI 118551 for the beta2 receptor relative to that for the beta1 receptor. The ability to estimate binding affinities of compounds using GPCR arrays was examined using a competitive binding assay with BT-NT and unlabeled neurotensin on NTR1 arrays. The estimated IC(50) value (2 nM) for neurotensin is in agreement with the literature; this agreement suggests that the receptor -G protein complex is preserved in the microspot. This first ever demonstration of direct pin-printing of membrane proteins and ligand-binding assays thereof fills a significant void in protein microchip technology--the lack of practical microarray-based methods for membrane proteins. PMID- 11890762 TI - Method for detection of single-base mismatches using bimolecular beacons. AB - This paper describes a method for the detection of single-base mismatches using DNA microarrays in a format that does not require labeling of the sample ("target") DNA. The method is based on disrupting fluorescence energy transfer (FRET) between a fluorophore attached to an immobilized DNA strand ("probe") and a quencher-containing sequence that is complementary except for an artificial mismatch (e.g. 5-nitroindole, 3-nitropyrole, or abasic site) at the site of interrogation. As the displacement of the FRET acceptor and hybridization of the unlabeled probe are bimolecular, the term "bimolecular beacons" is used to describe this approach. The analysis of a mismatch was based on differences in the amount of disruption in FRET upon hybridization of perfectly matched DNA targets and those containing single-base mismatches. Using this method and an oligonucleotide model system, A/C single-base mismatches were successfully discriminated at levels greater than that observed using surface-immobilized molecular beacons. The amount of discrimination was dependent on the identity of the artificial mismatch; greater discrimination was observed with 5-nitroindole (a "universal" base) than with an abasic site. G/T mismatches, considered to be particularly difficult to detect, were also successfully discriminated when quencher sequences containing 5-nitroindole were used. PMID- 11890763 TI - Molecular "compasses" and "gyroscopes". I. Expedient synthesis and solid state dynamics of an open rotor with a bis(triarylmethyl) frame. AB - We describe our efforts toward the preparation of materials built with molecules possessing topologies analogous to those of macroscopic compasses and gyroscopes. Samples of 1,4-bis(3,3,3-triphenylpropynyl)benzene (3) were prepared by a simple two-step procedure from triphenylmethyl chloride (1) and 1,4-diiodobenzene. The structure of compound 3 is such that the central phenylene can play the role of a gyroscope wheel while the two tritylpropynyl groups can act as an axle and shielding framework. Crystals of a benzene clathrate were characterized by single crystal X-ray diffraction and variable-temperature solid state NMR while their thermal stability was determined by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). The rotational dynamics of the phenylene group in the benzene clathrate and in desolvated samples were characterized in terms of a two-fold flipping process. Solid state rotational barriers of ca. 12.8 and 14.6 kcal/mol were estimated for the benzene clathrate and desolvated samples, respectively. PMID- 11890764 TI - Further insight in the photochemistry of DNA: structure of a 2-imidazolone (5-4) pyrimidone adduct derived from the mutagenic pyrimidine (6-4) pyrimidone photolesion by UV irradiation. AB - Pyrimidine (6-4) pyrimidone photoproducts represent one of the major mutagenic and carcinogenic class of DNA damage produced by UV exposure. At present, besides their conversion to their Dewar valence isomer, (6-4) photoproducts are generally believed to be photostable, and the observed biological properties of Paterno Buchi-derived photoproducts are, thus far, exclusively attributed to these two types of compounds. Using a model system (2) relevant to DNA photochemistry, we have observed that the 5'-base moiety of the (6-4) thymine dimer 3, under far-UV radiation, is able to undergo a ring contraction leading to a 2-oxoimidazoline, 1. This unprecedented secondary photochemical reaction constitutes the first report of a major photomodification affecting (6-4) photoproducts and strongly questions the biological stability of the (6-4) adducts under UV light with 2 imidazolone (5-4) pyrimidone adducts being possibly another source of endogenous DNA damage. PMID- 11890765 TI - Organocatalytic Michael cycloisomerization of bis(enones): the intramolecular Rauhut-Currier reaction. AB - The utilization of enones as latent enolates enables regioselective enolate formation from chemically robust presursors. In this communication, we report a catalytic Michael cycloisomerization of bis(enones) under Morita-Baylis-Hillman conditions. Upon exposure to 10 mol % tributylphosphine, bis(enone) substrates afford both five- and six-membered ring products. Notably, unsymmetrical bis(enones) possessing sufficient steric or electronic bias yield single isomeric products. PMID- 11890767 TI - An experimental test of C-N bond twisting in the TICT state: syn-anti photoisomerization in 2-(N-methyl-N-isopropylamino)-5-cyanopyridine. AB - A previously untested essential consequence of the "TICT" (twisted internal charge transfer) interpretation of the nature of the "anomalous" excited state of p-dimethylaminobenzonitrile (1) and related compounds has now been verified: at 90 degreesC, the dual fluorescence of a desymmetrized analogue, 2-(N-methyl-N isopropylamino)-5-cyanopyridine (2), in methanol (MeOH) is accompanied by syn anti isomerization around the C-N bond, whereas its ordinary fluorescence in tetrahydrofuran is not. PMID- 11890766 TI - The vinylogous intramolecular Morita-Baylis-Hillman reaction: synthesis of functionalized cyclopentenes and cyclohexenes with trialkylphosphines as nucleophilic catalysts. AB - The development of the vinylogous intramolecular Morita-Baylis-Hillman reaction for the synthesis of functionalized cyclopentenes and cyclohexenes is described. The reaction involves the trialkyphosphine-catalyzed cyclization of 1,6- or 1,7 diactivated 1,5-hexadienes or 1,6-heptadienes, containing carboxyaldehyde, methyl ketone, or methoxycarbonyl as the olefin activating groups. A representative example of this reaction is the Me(3)P-catalyzed cyclization of 1a in tert-amyl alcohol, which provides the substituted cyclopentene 2a in 95% yield and with 97:3 regioselectivity. PMID- 11890768 TI - SERS detection of the vibrational Stark effect from nitrile-terminated SAMs to probe electric fields in the diffuse double-layer. AB - The vibrational Stark effect is observed in the surface-enhanced Raman scattering spectra of self-assembled monolayers functionalized with pendant nitrile groups. Stark tuning of the nitrile-stretching frequency serves as a localized probe of the electric field in the diffuse double layer of a SAM-modified electrochemical interface. Stark-tuning rates at low ionic strength correspond to reasonable values of the local electric (E) field in the double layer. The nitrile stretching frequency converges on its isotropic value at applied potentials approaching the PZC, which indicates that Stark-tuning of the frequency is a direct probe of the E field at the interface. Loss of the local electric field at high ionic strengths shows that the probe responds to changes in the Debye length of the double layer. The results demonstrate the applicability of this electric field probe for characterizing the diffuse double-layer region. PMID- 11890769 TI - CsMBi(3)Te(6) and CsM(2)Bi(3)Te(7) (M = Pb, Sn): new thermoelectric compounds with low-dimensional structures. AB - The new materials CsPbBi(3)Te(6) and CsPb(2)Bi(3)Te(7) were discovered through reactions of CsBi(4)Te(6) with PbTe, whereas the isostructural materials CsSnBi(3)Te(6) and CsSn(2)Bi(3)Te(7) were discovered through corresponding reactions with SnTe. The compounds can also be prepared from stoichiometric mixtures of Cs(2)Te, Pb (Sn), Bi, and Te. The crystal structures show a layered architecture of NaCl-type slabs alternating with layers of Cs atoms. This group of compounds offers a new quaternary system, Cs-M-Bi-Te (M = Pb and Sn), available for thermoelectric investigations, including fine-tuning of compositions and doping. PMID- 11890770 TI - Methine (CH) transfer via a chlorine atom abstraction/benzene-elimination strategy: molybdenum methylidyne synthesis and elaboration to a phosphaisocyanide complex. AB - Methine (CH) transfer to an open coordination site was achieved in one pot by titanium(III) abstraction of Cl from 7-chloronorbornadiene, radical capture by Mo, and benzene extrusion. This efficient Mo methylidyne synthesis permitted elaboration to an anionic phosphaisocyanide derivative upon deprotonation, functionalization with dichlorophenylphosphine, and ultimate reduction. PMID- 11890771 TI - Nanoscale molecular patterns fabricated by using scanning near-field optical lithography. AB - Nanometer-scale patterns have been created in self-assembled monolayers by using a scanning near-field optical microscope coupled to an ultra-violet laser emitting light at a wavelength of 244 nm. Sharp, chemically well-defined features with dimensions as small as 40 nm have been created routinely, and on occasions line widths of 25 nm (lambda/10) have been achieved. Because of the wide range of photochemical methods available for surface derivatization, this approach promises to provide a flexible and versatile route to the generation of molecular and biological nanostructures for a wide range of applications. PMID- 11890772 TI - Catalytic oxidation by a carboxylate-bridged non-heme diiron complex. AB - The synthesis of a sterically hindered di(mu-carboxylato)diiron(II) complex bearing terminal N,N',N"-trimethyl-1,4,7-triazacyclononane (Me(3)TACN) ligands and its reaction with dioxygen to afford a (mu-oxo)di(mu-carboxylato)diiron(III) complex are described. Both compounds initiate catalytic oxo transfer with O(2) as the terminal oxidant, converting phosphines to phosphine oxides, dimethyl sulfide to dimethyl sulfoxide, and dibenzylamine to benzaldehyde. Triphenylphosphine is oxidized to triphenylphosphine oxide with a turnover number of >2000 mol.P/mol.cat. PMID- 11890773 TI - Intramolecular Forster energy transfer in a dendritic system at the single molecule level. AB - The photophysics of a dendrimer containing four donor chromophores and one acceptor chromophore are studied at the single-molecule level. Upon excitation of the donors exclusive acceptor emission is observed due to efficient Forster energy transfer. For 70% of the molecules donor emission is observed after bleaching of the acceptor, leading to a reduction of the Forster energy transfer efficiency. Furthermore, we demonstrate that in this molecular system the donor chromophores do not bleach by a triplet-sensitized photooxidation. PMID- 11890774 TI - Catalytic asymmetric direct alpha-amination reactions of 2-keto esters: a simple synthetic approach to optically active syn-beta-amino-alpha-hydroxy esters. AB - The catalytic enantioselective direct alpha-amination reaction of 2-keto esters with easily available azo dicarboxylates as the nitrogen source and chiral bisoxazoline-copper(II) complexes as the catalyst is presented. The reactions proceed with excellent enantioselectivities and high yields. The scope of the reaction is demonstrated by the direct amination of a series of 2-keto esters having different substituent patterns. The reaction provides an easy catalytic route to optically active syn-beta-amino-alpha-hydroxy esters, as demonstrated for the synthesis of masked syn-beta-amino-alpha-hydroxy esters (as oxazolidinones) and N-Boc-syn-beta-amino-alpha-hydroxy esters. On the basis of the absolute configuration of the products a tentative model for the transition state is suggested. PMID- 11890776 TI - Synthesis of a novel, highly symmetric bis-oxa-bridged compound. AB - A highly oxygenated, novel pentacyclic bis-oxa-bridged compound 8 was synthesized with remarkable efficiency starting from readily available tetrachloro-5,5 dimethoxyclopentadiene and 1,4-cyclohexadiene in three steps. The ruthenium catalyzed oxidation of 2:1 bis-adduct 1 followed by a one-pot transformation of the resulting bis-alpha-diketone 3 furnished (after esterification) the title compound in an overall yield of 29.1%. The versatility of this simple method was further demonstrated with other norbornyl alpha-diketones to obtain the corresponding strained oxa-bridged derivatives. PMID- 11890775 TI - Distance-dependent activation energies for hole injection from protonated 9-amino 6-chloro-2-methoxyacridine into duplex DNA. AB - The recent investigation of the apparently anomalous attenuation factor (beta > 1.5 A(-1)) for photoinduced hole injection into DNA duplexes modified by protonated 9-amino-6-chloro-2-methoxyacridine (X+) led to the conclusion that in addition to the electronic couplings, the activation energy must also be distance dependent. In this communication we report the verification of this postulate by direct measurements of the activation energies for a series of (X+)-modified DNA duplexes which sample an appreciable range of donor-acceptor distances (approximately 4-11 A). The resulting changes in thermal activation energy can be explained within the framework of a distance-dependent reorganization energy. PMID- 11890777 TI - Tracking alignment from the moment of inertia tensor (TRAMITE) of biomolecules in neutral dilute liquid crystal solutions. AB - NMR residual dipolar couplings between couple of nuclei PQ, (1)D(PQ), measured on neutral dilute liquid crystal solutions, provide valuable long-range structural information of biomolecules. An accurate and simple method for the prediction of the alignment produced as consequence of sterical interactions between the solute and the bicelles is proposed called TRacking Alignment from Moment of Inertia TEnsor--TRAMITE. The method use the information encoded in the moment of inertia of the molecules to calculate the orientation tensor and predict the (1)D(PQ) values. Examples on proteins and oligosaccharides are presented which cover a wide range of sizes and shapes, along with a scheme for the application of the method to the analysis of flexible molecules. PMID- 11890778 TI - Transition metal-coated nanoparticle films: vibrational characterization with surface-enhanced Raman scattering. AB - The utilization of surface-attached gold nanoparticles as templates for generating Pt-group particles displaying near-optimal surface-enhanced Raman scattering (SERS) characteristics is described. Essentially epitaxial transition metal coatings down to the monolayer level can be prepared, most readily by the spontaneous replacement of an electrochemically deposited copper layer by the desired Pt-group metal. The and essentially pinhole-free nature of the coated nanoparticles is demonstrated from the form of the SER spectra for chemisorbed carbon monoxide and ethylene. The potential of the present strategy for synthesizing relatively monodispersed "core-shell" nanoparticles using a myriad of coating materials, also displaying SERS activity, is pointed out. PMID- 11890779 TI - A general stereocontrolled, convergent synthesis of oligoprenols that parallels the biosynthetic pathway. AB - A solution is reported to the classic unsolved problem of stereoselective synthesis of all-E oligoprenols, such as E-farnesylfarnesol, by a cationic coupling analogous to the biosynthetic pathway. The simplicity and efficacy of the method, which is outlined in Scheme 1, are demonstrated by the synthesis of a series of all-E oligoprenols from C(20) to C(35) in uniformly excellent overall yield. The success of the approach is due not only to the highly E stereoselective C-C coupling that forms the oligoprenyl chain but also to the development of efficient syntheses of allylic secondary silanes and E-oligoprenal acetals, and to a selective allylic demethoxylation reaction. PMID- 11890780 TI - A novel chiral sulfonium yilde: highly enantioselective synthesis of vinylcyclopropanes. AB - A newly designed chiral sulfonium allylide, generated in situ from the corresponding sulfonium salt in the presence of KOBu(t), reacted with alpha,beta unsaturated esters, ketones, amides, and nitriles to afford trans-2-silylvinyl trans-3-substituted cyclopropyl esters, ketones, amides, and nitriles with outstanding diastereoselectivity and excellent enantioselectivity in good to high yields. A mechanistic rationale is proposed. PMID- 11890781 TI - Hydrotris(pyrazolyl)borate metallacycles: conversion of a late-metal metallacyclopentene to a stable metallacyclopentadiene-alkene complex. AB - The bis(ethene) complex [(Tp)Ir(C(2)H(4))(2)] (3) undergoes reaction with dimethyl acetylenedicarboxylate (DMAD) in acetonitrile solvent at 60 degrees C to give the trispyrazolylborate metallacyclopent-2-ene complex [(Tp)Ir (CH(2)CH(2)C(CO(2)Me)=C(CO(2)Me))(NCMe)] (4). Spectroscopic analysis of a room temperature reaction between 3 and DMAD in acetonitrile-d(3) provides evidence for the formation of an eta(2)-alkene/eta(2)-alkyne intermediate on the path to 4. The reaction of 3 with DMAD in THF solvent leads to the formation of the THF ligated metallacyclopent-2-ene complex [(Tp)Ir(CH(2)CH(2)C(CO(2)Me)=C(CO(2)Me))(THF)] (5), which undergoes further reaction with DMAD at 60 degrees C in benzene to give [(Tp)Ir(C(CO(2)Me)=C(CO(2)Me)C(CO(2)Me)=C(CO(2)Me))(eta(2)-CH(2)=CH(2))] (6). Complex 4 was structurally characterized by X-ray crystallography. PMID- 11890782 TI - Corynebactin and enterobactin: related siderophores of opposite chirality. AB - Most species of bacteria employ siderophores to acquire iron. The chirality of the ferric siderophore complex plays an important role in cell recognition, uptake, and utilization. Corynebactin, isolated from Gram-positive bacteria, is structurally similar to enterobactin, a well known siderophore isolated from Gram negative bacteria, but contains L-theronine instead of L-serine in the trilactone backbone. Corynebactin also contains a glycine spacer unit in each of the chelating arms. A hybrid analogue (serine-corynebactin) has been synthesized. The chirality and relative conformational stability of the three ferric complexes of enterobactin, corynebactin, and the hybrid has been investigated. In contrast to enterobactin, corynebactin assumes a Lambda configuration. However, the ferric serine-corynebactin hybrid forms a racemic mixture, only slightly favoring the Lambda conformation. PMID- 11890783 TI - Ultrafast bidirectional dihydroazulene/vinylheptafulvene (DHA/VHF) molecular switches: photochemical ring closure of vinylheptafulvene proven by a two-pulse experiment. AB - The photochromic couple dihydroazulene/vinylheptafulvene (DHA/VHF) is an established system for molecular switching. The photoinduced ring opening of the thermally stable dihydroazulene proceeds in the picosecond time regime with subsequent rapid relaxation to the electronic ground state. So far it was not possible to reverse the reaction photochemically. It was always found that the back reaction proceeds thermally on a longer time scale. In the case of the cyanophenyl-DHA derivative utilized in the present study, this is accounted to s cis/s-trans isomerization of the primarily formed s-cis VHF. Since this particular isomerization takes much longer than nanoseconds, we now successfully applied a second visible femtosecond pulse subsequent to the initial UV pulse (25 ps delay) achieving ring closure of the primarily formed s-cis VHF. The now bidirectional photoswitching was monitored by the changes in the cw spectrum. As a result, the DHA/VHF system is found to be a multifold switchable system by itself: it is both a very fast photoreversible switch and a photochemical/thermal switch with a thermal lock mode. PMID- 11890784 TI - A new strategy for caging proteins regulated by kinases. AB - A strategy has been developed for caging proteins that are endogenously regulated by phosphorylation. A key phosphorylatable serine in cofilin, an F-actin cleaving protein, was replaced with a nonphosphorylatable cysteine. The latter conversion ensures that the protein is no longer regulated by endogenous protein kinases. The cysteine residue was subsequently covalently modified with a negatively charged caging moiety, which electrostatically mimics the natural serine phosphate present in the inactive wild-type protein. Photoremoval of the cage generates an active protein, which cannot be switched off by endogenous protein kinases. Caged cofilin, and its irradiated counterpart, display the anticipated F actin depolymerization and severing activities. PMID- 11890785 TI - [Ph(2)Bi-(Ge(9))-BiPh(2)](2-): a deltahedral Zintl ion functionalized by exo bonded ligands. AB - The title anion was synthesized by a reaction of nido-Ge(9)(4-), made from K(4)Ge(9) dissolved in ethylenediamine and 2,2,2-crypt(4,7,13,16,21,24-hexaoxa 1,10-diazabicyclo[8.8.8]hexacosane), with BiPh(3). It was structurally characterized in (K-2,2,2-crypt)(2)[Ge(9)(BiPh(2))(2)].en which was crystallized from the solution. The anion is a monocapped square antiprism of Ge(9) with two diphenylbismuth ligands exo-bonded to opposite vertexes of the open face of the cluster. This is the first example where covalently exo-bonded ligands are attached to a deltahedral cluster that can exist without them as well. PMID- 11890786 TI - Sensitive and rapid analysis of protein palmitoylation with a synthetic cell permeable mimic of SRC oncoproteins. AB - The localization of oncogenic Src and Ras proteins to cellular plasma membranes is critical for the proliferation of specific cancers. In addition to other lipid modifications, these proteins require posttranslational palmitoylation of specific cysteine residues by the enzyme palmitoyl acyltransferase (PAT) in order to be stably anchored at plasma membranes. Hence, the identification of inhibitors of protein palmitoylation has significant potential to define a new class of antitumor agents. However, studies of protein palmitoylation have been hindered by the dynamic and reversible nature of cysteine acylation and the lack of sensitive and convenient assays of PAT activity. To facilitate the rapid identification of compounds that affect protein palmitoylation, we report the solid-phase synthesis of a fluorescent cell-permeable palmitoyl acyltransferase substrate that mimics the N-terminus of Src family proteins. Metabolic radiolabeling and epifluorescence microscopy of Jurkat lymphocytes treated with this Src-mimetic lipopeptide revealed that this compound is palmitoylated intracellularly, which confers localization at cellular plasma membranes. Addition of the palmitoylation inhibitor 2-bromopalmitic acid to substrate treated cells blocked palmitoylation and diminished substrate-mediated plasma membrane fluorescence. Analysis of inhibition of palmitoylation by flow cytometry revealed that this fluorescent lipopeptide substrate represents a highly sensitive molecular probe of palmitoyl acyltransferase activity that enables unprecedented high-throughput assays of protein palmitoylation. PMID- 11890787 TI - Site-selective screening by NMR spectroscopy with labeled amino acid pairs. AB - A new method for site-selective screening by NMR is presented. The core of the new method is the dual amino acid sequence specific labeling technique. Amino acid X is labeled with (13)C and amino acid Y is labeled with (15)N. Provided only one XY pair occurs in the amino acid sequence, only one signal in the 1D carbonyl (13)C spectrum will display a splitting due to the (1)J(C'N) coupling. Using this labeling strategy it is possible to screen selectively for binding to a selected epitope without the need for sequence specific assignments. An HNCO spectrum (1D or 2D) can be used either directly as a screening experiment or indirectly to identify what signals to monitor in a 2D (1)H-(15)N correlation spectrum. Chemical shift perturbations upon addition of a potential ligand are easily detected even for large proteins due to the reduced spectral complexity resulting from the use of a selectively labeled sample. The new technique is demonstrated on the human adipocyte fatty acid binding protein FABP-4. Due to the reduced spectral complexity, the method should be applicable to larger proteins than are conventional methods. PMID- 11890788 TI - Quantum dot on a rope. AB - The conjugation of nanoparticles (NPs) typically yields supramolecular materials which are fairly rigid, and the electronic coupling between the NP and other structural units of these compounds is fixed by covalent bonds. Here, we report on a novel bichromophor system constructed from a quantum dot tethered to a semiconducting polymer, which demonstrates the possibility of the dynamic interunit coupling in the NP supramolecules. The NP bichromophoric system was made on the basis of the layer-by-layer assembled (LBL) films of an anionic polyelectrolyte with poly(p-phenylene ethynylene) backbone, aPPE, and poly(allylamine hydrochloride) PAH polycation. To conjugate CdTe NPs to the (aPPE/PAH)(m) LBL film, we took advantage of the reactive groups of NP stabilizer, that is, -COOH, and the aminogroups on PAH. Tethering of CdTe was accomplished by using poly(ethyleneglycole), PEG, chains with two reactive terminals such as t-BOC-NH-PEG-COO-NHS. The evidence for successful conjugation of NPs to the LBL films can be seen both in AFM images and in optical data. The latter also indicate that the light quanta emitted by the NPs originate from the light absorption of the polymer film, which proves the presence of the aPPE-->NP energy-transfer process. The average separation distance between the NPs tethered to the LBL films can be changed by altering the dielectric properties of the solvent affecting PEG tether coiling (water/alcohol mixture). The reduced emission intensity of aPPE was found to follow the extension of the PEG tether. The quenching of aPPE is reversible when the original composition of the solvent mixture is restored. Thus, CdTe-PEG-aPPE is an example of an organized NP system with tunable optical coupling. Variable electronic coupling offers a convenient structural platform for new nanotechnological devices for which spatial control translates into a higher level of sophistication. PEG molecules afford a wide variety of polymer chain configurations with different reactive terminals, which makes possible the preparation of diverse NP superstructures. PMID- 11890789 TI - Micelle-induced curvature in a water-insoluble HIV-1 Env peptide revealed by NMR dipolar coupling measurement in stretched polyacrylamide gel. AB - The structure of a water-insoluble fragment encompassing residues 282-304 of the HIV envelope protein gp41 is studied when solubilized by dihexanoyl phosphatidylcholine (DHPC) and by small bicelles, consisting of a 4:1 molar ratio of DHPC and dimyristoyl phosphatidylcholine (DMPC). Weak alignment with the magnetic field was accomplished in a stretched polyacrylamide gel, permitting measurement of one-bond (1)H-(15)N, (13)Ca-(13)C', and (13)C'-(15)N dipolar couplings, which formed the basis for determining the peptide structure. In both detergent systems, the peptide adopts an alpha-helical conformation from residue 4 through 18. In the presence of the DHPC micelles the helix is strongly curved towards the hydrophobic surface, whereas in the presence of bicelles a much weaker curvature in the opposite direction is observed. PMID- 11890790 TI - Molecular "wiring" enzymes in organized nanostructures. AB - We report on the "molecular wiring" efficiency of glucose oxidase in organized self-assembled nanostructures comprised of enzyme layers alternating with layers of an osmium-derivatized poly(allylamine) cationic polyelectrolyte, acting as redox relays. Varying the relative position of the active enzyme layer in nanostructures alternating active enzyme and inactive apoenzyme we have demonstrated that the specific rate of bimolecular FADH(2) oxidation ("wiring efficiency") is limited by the diffusion-like electron hopping mechanism in the multilayers. PMID- 11890791 TI - Spin surface crossing in chromium-mediated olefin epoxidation with O(2). AB - [CpCr(mu-Cl)Cl](2) reacted with dioxygen (O(2)) to produce CpCr(O)Cl(2) (1), which has been structurally characterized. Although 1 oxidized PPh(3) and 1,4 cyclohexadiene catalytically, it did not epoxidize olefins. DFT calculations have been performed on the system to characterize the potential energy surface for the epoxidation of ethylene and, in particular, the consequences of the crossing from the doublet surface of the starting materials to the quartet surface of the product (i.e. a chromium(III) epoxide adduct). These calculations suggested that "spin-blocking" was not a significant problem and that the reaction of CpCr(O)Cl(2) (3) with ethylene should have a lower activation barrier. On the basis of this computational prediction, 3 was prepared; it was found to epoxidize olefins stoichiometrically. PMID- 11890792 TI - A direct method for the conversion of terminal epoxides into gamma-butanolides. AB - A new and efficient process for the conversion of terminal epoxides to gamma butanolides is described involving Lewis acid promoted epoxide ring-opening by 1 morpholino-2-trimethylsilyl acetylene. Addition of a terminal epoxide to a solution of the ynamine and boron trifluoride diethyl etherate in dichloromethane at 0 degrees C rapidly affords a cyclic keteneaminal that can be hydrolyzed and protodesilylated under mild conditions to provide the corresponding gamma butanolide in high yield. The net transformation is equivalent to an acetate enolate opening of terminal epoxides. The formation of a cyclic keteneaminal as the direct addition product was observed by monitoring of the reaction by IR and NMR spectroscopy. Functionalized gamma-lactones were prepared by the interception of the reactive cyclic keteneaminal prior to hydrolysis. Reactions with enantiomerically enriched terminal epoxides provide the corresponding gamma butanolides without loss of optical activity. The compatibility of the present methodology with a wide range of functional groups is noteworthy. PMID- 11890793 TI - The first general enantioselective catalytic Diels-Alder reaction with simple alpha,beta-unsaturated ketones. AB - The first general approach to enantioselective catalysis of the Diels-Alder reaction with simple ketone dienophiles has been accomplished. The use of iminium catalysis has enabled enantioselective access to a fundamental Diels-Alder reaction variant that has previously been unavailable using chiral Lewis acid catalysis. A new chiral amine catalyst has been developed that allows a variety of monodentate cyclic and acyclic ketones to successfully participate in enantioselective [4 + 2] cycloadditions. A wide spectrum of cyclic and acyclic diene substrates can also be accommodated in this new organocatalytic transformation. A computational model is provided that is in accord with the sense of enantioinduction observed for all reactions conducted during the course of this study. PMID- 11890794 TI - High-resolution X-ray structure of an acyl-enzyme species for the class D OXA-10 beta-lactamase. AB - Beta-lactamases are resistance enzymes for beta-lactam antibiotics. These enzymes hydrolyze the beta-lactam moieties of these antibiotics, rendering them inactive. Of the four classes of known beta-lactamases, the enzymes of class D are the least understood. We report herein the high-resolution (1.9 A) crystal structure of the class D OXA-10 beta-lactamase inhibited by a penicillanate derivative. The structure provides evidence that the carboxylated Lys-70 (a carbamate) is intimately involved in the mechanism of the enzyme. PMID- 11890795 TI - Noncovalent domino effect on helical screw sense of chiral peptides possessing C terminal chiral residue. AB - Recently, a novel chiral intermolecular interaction was found in an N-deprotected achiral nonapeptide that undergoes the predominance of one-handed screw sense through the addition of chiral small carboxylic acid (Inai, Y.; Tagawa, K.; Takasu, A.; Hirabayashi, T.; Oshikawa, T.; Yamashita, M. J. Am. Chem. Soc. 2000, 122, 11731). We here clarify to what extent such noncovalent chiral domino effect affects the helical screw sense of an N-deprotected chiral peptide. Two chiral peptides consisting of C-terminal L-Leu (1) or L-Leu(2) (2) and the preceding achiral helical octapeptide segment were employed. NMR and IR spectroscopy, and energy calculation indicated that both peptides adopt a helical conformation in chloroform. Peptide 1 showed a small excess of a left-handed screw sense for the achiral helical octapeptide, but peptide 2 strongly preferred a right-handed screw sense. The addition of chiral Boc amino acid to a chloroform solution of peptide 1, depending on its chirality, underwent a unique helix-to-helix transition or led to remarkable stabilization of the original left-handed screw sense. Peptide 2 retained the original right-handed screw sense on addition of chiral Boc-amino acid, but its helical stability changed to some extent depending on its added chirality. Therefore, the importance of noncovalent domino effect for controlling the helical screw sense or helical stability of a chiral peptide has been demonstrated here for the first time. In addition, we here have presented a unique system that both N-terminal noncovalent and C-terminal covalent domino effects operate simultaneously on the helical screw sense of a single achiral segment and have compared both powers for inducing the screw sense bias. PMID- 11890796 TI - Effect of sequence on peptide geometry in 5-tert-butylprolyl type VI beta-turn mimics. AB - The influence of sequence on turn geometry was examined by incorporating (2S,5R) 5-tert-butylproline (5-(t)BuPro) into a series of dipeptides and tetrapeptides. (2S,5R)-5-tert-Butylproline and proline were respectively introduced at the C terminal residue of N-acetyl dipeptide N'-methylamides 1 and 2. The conformational analysis of these analogues was performed using NMR and CD spectroscopy as well as X-ray diffraction to examine the factors that control the prolyl amide (in this text, the term "prolyl amide" refers to the tertiary amide composed of the pyrrolidine nitrogen of the prolyl residue and the carbonyl of the N-terminal residue) equilibrium and stabilize type VI beta-turn conformation. The high cis-isomer population with aromatic residues N-terminal to proline was shown to result from a stacking interaction between the partial positive charged prolyl amide nitrogen and the aromatic pi-system as seen in the crystal structure of 1c. The effect of sequence on the prolyl amide equilibrium of 5-(t)BuPro tetrapeptides (Ac-Xaa-Yaa-5-(t)BuPro-Zaa-XMe, 13 and 14) was studied by varying the amino acids at the Xaa, Yaa, and Zaa positions. High (>80%) cis-isomer populations were obtained with alkyl groups at the Xaa position, an aromatic residue at the Yaa position, and either an alanine or a lysine residue at the Zaa position of the 5-(t)BuPro-tetrapeptide methyl esters in water. Tetrapeptides Ac Ala-Phe-5-(t)BuPro-Zaa-OMe (Zaa = Ala, Lys), 14d and 14f, with high cis-isomer content adopted type VIa beta-turn conformations as shown by their NMR and CD spectra. Although a pattern of amide proton temperature coefficient values indicative of a hairpin geometry was observed in peptides 14d and 14f, the value magnitudes did not indicate strong hydrogen bonding in water. PMID- 11890797 TI - Definitive evidence for monoanionic binding of 2,3-dihydroxybiphenyl to 2,3 dihydroxybiphenyl 1,2-dioxygenase from UV resonance Raman spectroscopy, UV/Vis absorption spectroscopy, and crystallography. AB - Ultraviolet resonance Raman spectroscopy (UVRRS), electronic absorption spectroscopy, and X-ray crystallography were used to probe the nature of the binding of 2,3-dihydroxybiphenyl (DHB) to the extradiol ring-cleavage enzyme, 2,3 dihydroxybiphenyl 1,2-dioxygenase (DHBD; EC 1.13.11.39). The lowest lying transitions in the electronic absorption spectrum of DHBD-bound DHB occurred at 299 nm, compared to 305 nm for the monoanionic DHB species in buffer. In contrast, the corresponding transitions in neutral and dianionic DHB occurred at 283 and 348 nm, respectively, indicating that DHBD-bound DHB is monoanionic. These binding-induced spectral changes, and the use of custom-designed optical fiber probes, facilitated UVRR experiments. The strongest feature of the UVRR spectrum of DHB was a Y8a-like mode around 1600 cm(-1), whose position depended strongly on the protonation state of the DHB. In the spectrum of the DHBD-bound species, this feature occurred at 1603 cm(-1), as observed in the spectrum of monoanionic DHB. Raman band shifts were observed in deuterated solvent, ruling out dianionic binding of the substrate. Thus, the electronic absorption and UVRRS data demonstrate that DHBD binds its catecholic substrate as a monoanion, definitively establishing this feature of the proposed mechanism of extradiol dioxygenases. This conclusion is supported by a crystal structure of the DHBD:DHB complex at 2.0 A resolution, which suggests that the substrate's 2-hydroxyl substituent, and not the 3-hydroxyl group, deprotonates upon binding. The structural data also show that the aromatic rings of the enzyme-bound DHB are essentially orthogonal to each other. Thus, the 6 nm blue shift of the transition for bound DHB relative to the monoanion in solution could indicate a conformational change upon binding. Catalytic roles of active site residues are proposed based on the structural data and previously proposed mechanistic schemes. PMID- 11890798 TI - Collagen stability: insights from NMR spectroscopic and hybrid density functional computational investigations of the effect of electronegative substituents on prolyl ring conformations. AB - Collagen-like peptides of the type (Pro-Pro-Gly)(10) fold into stable triple helices. An electron-withdrawing substituent at the H(gamma)(3) ring position of the second proline residue stabilizes these triple helices. The aim of this study was to reveal the structural and energetic origins of this effect. The approach was to obtain experimental NMR data on model systems and to use these results to validate computational chemical analyses of these systems. The most striking effects of an electron-withdrawing substituent are on the ring pucker of the substituted proline (Pro(i)) and on the trans/cis ratio of the Xaa(i-1)-Pro(i) peptide bond. NMR experiments demonstrated that N-acetylproline methyl ester (AcProOMe) exists in both the C(gamma)-endo and C(gamma)-exo conformations (with the endo conformation slightly preferred), N-acetyl-4(R)-fluoroproline methyl ester (Ac-4R-FlpOMe) exists almost exclusively in the C(gamma)-exo conformation, and N-acetyl-4(S)-fluoroproline methyl ester (Ac-4S-FlpOMe) exists almost exclusively in the C(gamma)-endo conformation. In dioxane, the K(trans/cis) values for AcProOMe, Ac-4R-FlpOMe, and Ac-4S-FlpOMe are 3.0, 4.0, and 1.2, respectively. Density functional theory (DFT) calculations with the (hybrid) B3LYP method were in good agreement with the experimental data. Computational analysis with the natural bond orbital (NBO) paradigm shows that the pucker preference of the substituted prolyl ring is due to the gauche effect. The backbone torsional angles, phi and psi, were shown to correlate with ring pucker, which in turn correlates with the known phi and psi angles in collagen-like peptides. The difference in K(trans/cis) between AcProOMe and Ac-4R-FlpOMe is due to an n-->pi interaction associated with the Burg-Dunitz trajectory. The decrease in K(trans/cis) for Ac-4S-FlpOMe can be explained by destabilization of the trans isomer because of unfavorable electronic and steric interactions. Analysis of the results herein along with the structures of collagen-like peptides has led to a theory that links collagen stability to the interplay between the pyrrolidine ring pucker, phi and psi torsional angles, and peptide bond trans/cis ratio of substituted proline residues. PMID- 11890799 TI - Synthesis and rearrangement of diphosphorus analogues of amidinium salts. AB - Transient diphosphinocarbocations IIP are generated either by addition of phosphenium salts to the stable [bis(diisopropylamino)phosphino](silyl)carbene or by chloride abstraction from C-phosphino-P-chloro phosphorus ylides. In contrast to their nitrogen anlogues (amidinium salts) IIN, which feature a planar 3-center 4p-electron system, calculations show that IIP should exist as IIPb, in which one phosphorus is planar, while the other remains pyramidal. With small substituents at phosphorus, derivatives of type IIP rearrange by a 1,3-shift of a phosphorus substituent to the other phosphorus center to give C-phosphoniophosphaalkenes. When bulky substituents are present at phosphorus, derivatives IIP undergo ring closure, giving rise to the corresponding cyclic valence isomers IIIP, in which the carbon atom bears a negative charge. Diphosphinocarbocations IIP can be trapped by acetonitrile giving regioselectively the corresponding [2+3] cycloadduct. PMID- 11890800 TI - CO(2)-expanded solvents: unique and versatile media for performing homogeneous catalytic oxidations. AB - The work summarized here demonstrates a new concept for exploiting dense phase CO(2), media considered to be "green" solvents, for homogeneous catalytic oxidation reactions. According to this concept, the conventional organic solvent medium used in catalytic chemical reactions is replaced substantially (up to 80 vol %) by CO(2), at moderate pressures (tens of bars), to create a continuum of CO(2)-expanded solvent media. A particular benefit is found for oxidation catalysis; the presence of CO(2) in the mixed medium increases the O(2) solubility by ca. 100 times compared to that in the neat organic solvent while the retained organic solvent serves an essential role by solubilizing the transition metal catalyst. We show that CO(2)-expanded solvents provide optimal properties for maximizing oxidation rates that are typically 1-2 orders of magnitude greater than those obtained with either the neat organic solvent or supercritical CO(2) as the reaction medium. These advantages are demonstrated with examples of homogeneous oxidations of a substituted phenol and of cyclohexene by molecular O(2) using transition metal catalysts, cobalt Schiff base and iron porphyrin complexes, respectively, in CO(2)-expanded CH(3)CN. PMID- 11890801 TI - Fluorescence enhancement of trans-4-aminostilbene by N-phenyl substitutions: the "amino conjugation effect". AB - The synthesis, structure, and photochemical behavior of the trans isomers of 4-(N phenylamino)stilbene (1c), 4-(N-methyl-N-phenylamino)stilbene (1d), 4-(N,N diphenylamino)stilbene (1e), and 4-(N-(2,6-dimethylphenyl)amino)stilbene (1f) are reported and compared to that of 4-aminostilbene (1a) and 4-N,N dimethylaminostilbene (1b). Results for the corresponding 3-styrylpyridine (2) and 2-styrylnaphthalene analogues (3) are also included. The introduction of N phenyl substituents to 4-aminostilbenes leads to a more planar ground-state geometry about the nitrogen atom, a red shift of the absorption and fluorescence spectra, and a less distorted structure with a larger charge-transfer character for the fluorescent excited state. Consequently, the N-phenyl derivatives 1c-e have low photoisomerization quantum yields and high fluorescence quantum yields at room temperature, in contrast to the behavior of 1a, 1b, and most unconstrained monosubstituted trans-stilbenes. The isomerization of 1c and 1d is a singlet-state process, whereas it is a triplet-state process for 1e, presumably due to a relatively higher singlet-state torsional barrier. The excited-state behavior of 1f resembles 1a and 1b instead of 1c-e as a consequence of the less planar amine geometry and weaker orbital interactions between the N-phenyl and the aminostilbene groups. Such an N-phenyl substituent effect is also found for 2 and 3 and thus appears to be general for stilbenoid systems. The nature of this effect can be described as an "amino conjugation effect". PMID- 11890802 TI - A Ru catalyzed divergence: oxidative cyclization vs cycloisomerization of bis homopropargylic alcohols. AB - During the course of investigating the development of catalytic reactions involving ruthenium vinylidene intermediates, a novel divergence of reactivity was discovered. The oxidative cyclization of bis-homopropargylic alcohols with Ru(+2) complexes as catalysts and N-hydroxysuccinimide as oxidant, which requires formation of a ruthenium vinylidene intermediate, is complicated by the simple electrophilically initiated direct attack of the hydroxyl group on a pi-complex of the alkyne and ruthenium. A catalytic system composed of CpRu[(p CH(3)O(6)H(4))(3)P](2)Cl and excess (p-CH(3)O-C(6)H(4))(3)P directs the reaction toward the oxidative cyclization to form delta-lactones in good yields. Significantly, a simple switch of catalyst to CpRu[(p-FC(6)H(4))(3)P](2)Cl redirects the reaction to a cycloisomerization to form dihydropyrans in good yields. The synthetic utility of the oxidative cyclization is illustrated by the synthesis of oviposition attractant pheromone of the mosquito Culex pipens. The utility of the cycloisomerization to dihydropyrans is demonstrated by an iterative process leading to the antiviral agent narbosine B. A rationale for this dramatic switch by simple ligand modification is proposed. PMID- 11890803 TI - Application of serine- and threonine-derived cyclic sulfamidates for the preparation of S-linked glycosyl amino acids in solution- and solid-phase peptide synthesis. AB - Cyclic sulfamidates were synthesized in 60% yield from L-serine and allo-L threonine, respectively. These sulfamidates reacted with a variety of unprotected 1-thio sugars in aqueous bicarbonate buffer (pH 8) to afford the corresponding S linked serine- and threonine-glycosyl amino acids with good diastereoselectivity (> or =97%) after hydrolysis of the N-sulfates. The serine-derived sulfamidate was incorporated into a simple dipeptide to generate a reactive dipeptide substrate that underwent chemoselective ligation with a 1-thio sugar to afford an S-linked glycopeptide. This sulfamidate was also incorporated into a peptide on a solid support in conjunction with solid-phase peptide synthesis. Chemoselective ligation of a 1-thio sugar with the cyclic sulfamidate was achieved on the solid support, followed by removal of the N-sulfate. Finally, the peptide chain of the resulting support-bound S-linked glycopeptide was extended using standard peptide synthesis procedures. PMID- 11890804 TI - Evidence for a nonradical pathway in the photoracemization of aryl sulfoxides. AB - Photolysis of (R(S),S(C))-1-deuterio-2,2-dimethylpropyl p-tolyl sulfoxide provides mainly (S(S),S(C))-1-deuterio-2,2-dimethylpropyl p-tolyl sulfoxide at low conversion, though the other two stereoisomers are formed to smaller extents. Thus, the predominant process leading to sulfur inversion yields only sulfur inversion, without inversion of the adjacent CHD stereogenic center. This is taken as evidence for a mechanism for photochemical epimerization of sulfoxides that does not involve homolytic alpha-cleavage chemistry. PMID- 11890805 TI - Chain epimerization during propylene polymerization with metallocene catalysts: mechanistic studies using a doubly labeled propylene. AB - The mechanisms of chain epimerization during propylene polymerization with methylaluminoxane-activated rac-(EBTHI)ZrCl(2) and rac-(EBI)ZrCl(2) catalysts (EBTHI = ethylenebis(eta(5)-tetrahydroindenyl); EBI = ethylenebis(eta(5) indenyl)) have been studied using specifically isotopically labeled propylene: CH(2)=CD(13)CH(3). These isospecific catalysts provide predominantly the expected [mmmm] pentads with [minus signCH(2)CD(13)CH(3)(-)] repeating units ((13)C NMR). Under relatively low propylene concentrations at 50 and 75 degreesC, where stereoerrors attributable to chain epimerization are prevalent, (13)C NMR spectra reveal (13)C-labeled methylene groups along the polymer main chain, together with [CD(13)CH(3)] units in [mmmr], [mmrr], and [mrrm] pentads and [CH(13)CH(3)] units in [mmmmmm] and [mmmmmr] heptads, as well as [mrrm] pentads. The isotopomeric regiomisplacements and stereoerrors are consistent with a mechanism involving beta-D elimination, olefin rotation and enantiofacial interconversions, and insertion to a tertiary alkyl intermediate [Zr-C(CH(2)D)((13)CH(3))P] (P = polymer chain), followed by the reverse steps to yield two stereoisomers of [Zr CHDCH((13)CH(3))P] and [Zr-(13)CH(2)CH(CH(2)D)P], as well as unrearranged [Zr CH(2)CD((13)CH(3))P]. The absence of observable [-CH(2)CH(13)CH(2)D-] in the [mrrm] pentad region of the (13)C NMR spectra provides evidence that an allyl/dihydrogen complex does not mediate chain epimerization. PMID- 11890806 TI - Asymmetric total synthesis of (-)-VM55599: establishment of the absolute stereochemistry and biogenetic implications. AB - The first asymmetric biomimetic total synthesis of VM55599 (13) has been achieved utilizing an intramolecular Diels-Alder cycloaddition as a key step. The synthetic material was utilized to elucidate the absolute stereochemistry of the natural product. The results are discussed in terms of a unified biogenesis of the paraherquamides and VM55599. PMID- 11890807 TI - Ammonium chloride-promoted four-component synthesis of pyrrolo[3,4-b]pyridin-5 one. AB - A novel multicomponent synthesis of 5-aminooxazole starting from simple and readily available inputs is described. Thus, simply heating a methanol solution of an aldehyde 3, an amine 4, and an alpha-isocyanoacetamide 5 provided the 5 aminooxazole (1) in good to excellent yield. The reaction of 1 with alpha,beta unsaturated acyl chloride 13 lead to the formation of pyrrolo[3,4-b]pyridin-5-one (2) in a single operation. A triple domino sequence, acylation/IMDA/retro-Michael cycloreversion, is involved in this new scaffold-generating reaction. After the observation that ammonium chloride can significantly accelerate the oxazole formation in toluene, a one-pot four-component synthesis of 2 is developed. PMID- 11890808 TI - Framework engineering by anions and porous functionalities of Cu(II)/4,4'-bpy coordination polymers. AB - A combination of framework-builder (Cu(II) ion and 4,4'-bipyridine (4,4'-bpy) ligand) and framework-regulator (AF(6) type anions; A = Si, Ge, and P) provides a series of novel porous coordination polymers. The highly porous coordination polymers ([Cu(AF(6))(4,4'-bpy)(2)].8H(2)O)(n)(A = Si (1a.8H(2)O), Ge (2a.8H(2)O)) afford robust 3-dimensional (3-D), microporous networks (3-D Regular Grid) by using AF(6)(2-) anions. The channel size of these complexes is ca. 8 x 8 A(2) along the c-axis and 6 x 2 A(2) along the a- or b-axes. When compounds 1a.8H(2)O or 2a.8H(2)O were immersed in water, a conversion of 3-D networks (1a.8H(2)O or 2a.8H(2)O) to interpenetrated networks ([Cu(4,4'-bpy)(2)(H(2)O)(2)].AF(6))(n)(A = Si (1b) and Ge (2b)) (2-D Interpenetration) took place. This 2-D interpenetrated network 1b shows unique dynamic anion-exchange properties, which accompany drastic structural conversions. When a PF(6)(-) monoanion instead of AF(6)(2)(-) dianions was used as the framework-regulator with another co-counteranion (coexistent anions), porous coordination polymers with various types of frameworks, ([Cu(2)(4,4'-bpy)(5)(H(2)O)(4)].anions.2H(2)O.4EtOH)(n)(anions = 4PF(6)(-) (3.2H(2)O.4EtOH), 2PF(6)(-) + 2ClO(4)(-) (4.2H(2)O.4EtOH)) (2-D Double Layer), ([Cu(2)(PF(6))(NO(3))(4,4'-bpy)(4)].2PF(6).2H(2)O)(n)(5.2PF(6).2H(2)O) (3 D Undulated Grid), ([Cu(PF(6))(4,4'-bpy)(2)(MeCN)].PF(6).2MeCN)(n)(6.2MeCN) (2-D Grid), and ([Cu(4,4'-bpy)(2)(H(2)O)(2)].PF(6).BF(4))(n) (7) (2-D Grid), were obtained, where the three modes of PF(6)(-) anions are observed. 5.2PF(6).2H(2)O has rare PF(6)(-) bridges. The PF(6)(-) and NO(3)(-) monoanions alternately link to the Cu(II) centers in the undulated 2-D sheets of [Cu(4,4'-bpy)(2)](n)() to form a 3-D porous network. The free PF(6)(-) anions are included in the channels. 6.2MeCN affords both free and terminal-bridged PF(6)(-) anions. 3.2H(2)O.4EtOH, 4.2H(2)O.4EtOH, and 7 bear free PF(6)(-) anions. All of the anions in 3.2H(2)O.4EtOH and 4.2H(2)O.4EtOH are freely located in the channels constructed from a host network. Interestingly, these Cu(II) frameworks are rationally controlled by counteranions and selectively converted to other frameworks. PMID- 11890809 TI - Formation of cis-enediyne complexes from rhenium alkynylcarbene complexes. AB - Dimerization of the alkynylcarbene complex Cp(CO)(2)Re=C(Tol)C(triple bond)CCH(3) (8) occurs at 100 degrees C to give a 1.2:1 mixture of enediyne complexes [Cp(CO)(2)Re](2)[eta(2),eta(2)-TolC(triple bond)CC(CH(3))=C(CH(3))C(triple bond)CTol] (10-Eand 10-Z), showing no intrinsic bias toward trans-enediyne complexes. The cyclopropyl-substituted alkynylcarbene complex Cp(CO)(2)Re=C(Tol)C(triple bond)CC(3)H(5) (11) dimerizes at 120 degrees C to give a 5:1 ratio of enediyne complexes [Cp(CO)(2)Re](2)[eta(2),eta(2)-TolC(triple bond)C(C(3)H(5))C=C(C(3)H(5))C(triple bond)CTol] (12-E and 12-Z); no ring expansion product was observed. This suggests that if intermediate A formed by a [1,1.5] Re shift and having carbene character at the remote alkynyl carbon is involved, then interaction of the neighboring Re with the carbene center greatly diminishes the carbene character as compared with that of free cyclopropyl carbenes. The tethered bis-(alkynylcarbene) complex Cp(CO)(2)Re=C(Tol)C(triple bond)CCH(2)CH(2)CH(2)C(triple bond)CC(Tol)= Re(CO)(2)Cp (13) dimerizes rapidly at 12 degrees C to give the cyclic cis-enediyne complex [Cp(CO)(2)Re](2)[eta(2),eta(2)-TolC(triple bond)CC(CH(2)CH(2)CH(2))=CC(triple bond)CTol] (15). Attempted synthesis of the 1,8-disubstituted naphthalene derivative 1,8-[Cp(CO)(2)Re=C(Tol)C(triple bond)C](2)C(10)H(6) (16), in which the alkynylcarbene units are constrained to a parallel geometry, leads to dimerization to [Cp(CO)(2)Re](2)(eta(2),eta(2)-1,2-(tolylethynyl)acenaphthylene] (17). The very rapid dimerizations of both 13 and 16 provide compelling evidence against mechanisms involving cyclopropene intermediates. A mechanism is proposed which involves rate-determining addition of the carbene center of A to the remote alkynyl carbon of a second alkynylcarbene complex to generate vinyl carbene intermediate C, and rearrangement of C to the enediyne complex by a [1,1.5] Re shift. PMID- 11890811 TI - Platinum chalcogenido MCM-41 analogues. High hexagonal order in mesostructured semiconductors based on Pt(2+) and [Ge(4)Q(10)](4-) (Q = S, Se) and [Sn(4)Se(10)](4-) adamantane clusters. AB - Highly periodic hexagonal honeycombs of platinum-germanium chalcogenide and platinum-tin selenide frameworks were prepared by linking corresponding [Ge(4)Q(10)](4)(-) (Q = S, Se) and [Sn(4)Se(10)](4)(-) clusters with Pt(2+) ions. The non-oxidic honeycombs designated as C(n)PyPtGeQ and C(n)PyPtSnSe were templated by the lyotropic liquid-crystalline phase of alkylpyridinium surfactant [C(n)H(2)(n)(+1)NC(5)H(5)]Br (C(n)PyBr) with n= 12, 14, 16, 18, 20, and 22. Although the materials are amorphous at the microscale, they have crystalline mesoporosity with well-ordered and aligned surfactant-filled cylindrical pores. In addition to high mesoscopic order, the pore-pore separation is adjustable with the surfactant chain length (i.e., value of n). The quality of these materials, as judged by the degree of hexagonal order, rivals or exceeds that reported for the highest quality MCM-41 silicates. The materials have the lowest band gap reported so far for mesostructured chalcogenides solids, in the range 1.5 < E(g)< 2.3 eV. The C(n)PyPtGeS analogues show intense photoluminescence at 77 K when excited with light above the band gap. PMID- 11890810 TI - Electro-nuclear double resonance spectroscopic evidence for a hydroxo-bridge nucleophile involved in catalysis by a dinuclear hydrolase. AB - Despite the current availability of several crystal structures of purple acid phosphatases, to date there is no direct evidence for solvent-derived ligands occupying terminal positions in the active enzyme. This is of central importance, because catalysis has been shown to proceed through the direct attack on a metal bound phosphate ester by a metal-activated solvent-derived moiety, which has been proposed to be either (i) a hydroxide ligand terminally bound to the ferric center or (ii) a bridging hydroxide. In this work we use (2)H Q-band (35 GHz) pulsed electron-nuclear double resonance (ENDOR) spectroscopy to identify solvent molecules coordinated to the active mixed-valence (Fe(3+)Fe(2+)) form of the dimetal center of uteroferrin (Uf), as well as to its complexes with the anions MoO(4), AsO(4), and PO(4). The solvent-derived coordination of the dinuclear center of Uf as deduced from ENDOR data includes a bridging hydroxide and a terminal water/hydroxide bound to Fe(2+) but no terminal water/hydroxide bound to Fe(3+). The terminal water is lost upon anion binding while the hydroxyl bridge remains. These results are not compatible with a hydrolysis mechanism involving a terminal Fe(3+)-bound nucleophile, but they are consistent with a mechanism that relies on the bridging hydroxide as the nucleophile. PMID- 11890812 TI - S-alkylation and S-amination of methyl thioethers--derivatives of closo [B(12)H(12)](2-). synthesis of a boronated phosphonate, gem-bisphosphonates, and dodecaborane-ortho-carborane oligomers. AB - A variety of S-alkylated products was prepared by alkylation of methyl thioethers [MeSB(12)H(11)](2-) (5), [1-(MeS)-2(7,12)-(Me(2)S)B(12)H(10)](-) (6-8), and [1,2(7,12)-(MeS)(2)B(12)H(10)](2-) (9-11) with alkyl halides and tosylates in acetonitrile. Since these methyl thioethers can be prepared easily in B-10 enriched form on a large scale and due to their chemical versatility, they are potentially very attractive boron entities for the design and synthesis of therapeutics for boron neutron capture therapy of cancer. It was found that alkylation of 6-8 can be complicated by an equilibrium which establishes between, on the one hand, one of the former species and, on the other hand, 1,2(7,12) (Me(2)S)(2)B(12)H(10) (2-4) and [1,2(7,12)-(MeS)(2)B(12)H(10)](2-) (9-11). A boronated phosphonate 1-(MeS(CH(2))(4)P(O)(OEt)(2))-7-(Me(2)S)B(12)H(10) (14g) and a gem-bisphosphonate 1-(MeS(CH(2))(3)CH[P(O)(OEt)(2)](2))-7 (Me(2)S)B(12)H(10) (14h) were prepared from thioether 7 and the corresponding iodide and tosylate, respectively, and subsequently converted to their sodium salts. The propargyl sulfonium salts obtained by alkylation of thioethers 7, 8, 10, and 11 with propargyl bromide have been further converted to two- and three cage oligomers containing both ortho-carborane and dodecaborane moieties. Methyl thioethers derived from closo-[B(12)H(12)](2-) are excellent participants in Michael addition reactions in the presence of a strong acid. The sulfonium salts with tertiary alkyl and vinyl substituents have been prepared by this method. Methyl thioethers 5-11 react with hydroxylamine-O-sulfonate yielding the corresponding aminosulfonium salts, albeit in lower yields as compared to those in the alkylation reactions. Several derivatives of methyl thioethers 5-11 have been characterized by single-crystal X-ray diffraction. PMID- 11890814 TI - Hydrothermal oxidation-reduction methods for the preparation of pure and single crystalline alunites: synthesis and characterization of a new series of vanadium jarosites. AB - Three new redox-based, hydrothermal, synthetic methods have been developed for the preparation of a new series of jarosites, AV(3)(OH)(6)(SO(4))(2) (A = Na(+), K(+), Rb(+), Tl(+), and NH(4)(+)), in high purity and in single crystalline form. The V(3+) jarosites have been characterized by single-crystal X-ray and elemental analysis, and by infrared and electronic absorption spectroscopy. The synthetic methods employed here represent a new approach for the preparation of the jarosite class of compounds, which for the past several decades, have been notoriously difficult to prepare in pure form. To demonstrate the impact of our new synthetic techniques on the magnetic properties of jarosites, the V(3+) jarosites were also prepared according to the nonredox techniques used over the past 30 years. A comparative study of these samples and those prepared by our new synthetic methods reveals widely divergent magnetic properties, thus pointing to the importance of the new redox synthetic methods to future magnetism studies of jarosite compounds. PMID- 11890813 TI - Genesis of nanostructured, magnetically tunable ceramics from the pyrolysis of cross-linked polyferrocenylsilane networks and formation of shaped macroscopic objects and micron scale patterns by micromolding inside silicon wafers. AB - The ability to form molded or patterned metal-containing ceramics with tunable properties is desirable for many applications. In this paper we describe the evolution of a ceramic from a metal-containing polymer in which the variation of pyrolysis conditions facilitates control of ceramic structure and composition, influencing magnetic and mechanical properties. We have found that pyrolysis under nitrogen of a well-characterized cross-linked polyferrocenylsilane network derived from the ring-opening polymerization (ROP) of a spirocyclic [1]ferrocenophane precursor gives shaped macroscopic magnetic ceramics consisting of alpha-Fe nanoparticles embedded in a SiC/C/Si(3)N(4) matrix in greater than 90% yield up to 1000 degrees C. Variation of the pyrolysis temperature and time permitted control over the nucleation and growth of alpha-Fe particles, which ranged in size from around 15 to 700 A, and the crystallization of the surrounding matrix. The ceramics contained smaller alpha-Fe particles when prepared at temperatures lower than 900 degrees C and displayed superparamagnetic behavior, whereas the materials prepared at 1000 degrees C contained larger alpha Fe particles and were ferromagnetic. This flexibility may be useful for particular materials applications. In addition, the composition of the ceramic was altered by changing the pyrolysis atmosphere to argon, which yielded ceramics that contain Fe(3)Si(5). The ceramics have been characterized by a combination of physical techniques, including powder X-ray diffraction, TEM, reflectance UV vis/near-IR spectroscopy, elemental analysis, XPS, SQUID magnetometry, Mossbauer spectroscopy, nanoindentation, and SEM. Micromolding of the spirocyclic [1]ferrocenophane precursor within soft lithographically patterned channels housed inside silicon wafers followed by thermal ROP and pyrolysis enabled the formation of predetermined micron scale designs of the magnetic ceramic. PMID- 11890815 TI - Magnetic properties of a homologous series of vanadium jarosite compounds. AB - Redox-based, hydrothermal synthetic methodologies have enabled the preparation of a new series of stoichiometrically pure jarosites of the formula, AV(3)(OH)(6)(SO(4))(2) with A = Na(+), K(+), Rb(+), Tl(+), and NH(4)(+). These jarosites represent the first instance of strong ferromagnetism within a Kagome layered framework. The exchange interaction, which is invariant to the nature of the A(+) ion (theta(CW) approximately equal to +53(1) K), propagates along the d(2) magnetic sites of the triangular Kagome lattice through bridging hydroxyl groups. An analysis of the frontier orbitals suggests this superexchange pathway to possess significant pi-orbital character. Measurements on a diamagnetic host jarosite doped with magnetically dilute spin carriers, KGa(2.96)V(0.04)(OH)(6)(SO(4))(2), reveal significant single-ion anisotropy for V(3+) ion residing in the tetragonal crystal field. This anisotropy confines the exchange-coupled moments to lie within the Kagome layer. Coupling strengths are sufficiently strong to prevent saturation of the magnetization when an external field is applied orthogonal to the Kagome layer. Antiferromagnetic ordering of neighboring ferromagnetic Kagome layers becomes dominant at low temperatures, characteristic of metamagnetic behavior for the AV(3)(OH)(6)(SO(4))(2) jarosites. This interlayer exchange coupling decreases monotonically with increasing layer spacing along the series, A = Na(+), K(+), Rb(+), NH(4)(+), and Tl(+), and it may be overcome by the application of external field strengths in excess of approximately 6 kOe. PMID- 11890816 TI - Product binding to the diiron(III) and mixed-valence diiron centers of methane monooxygenase hydroxylase studied by (1,2)H and (19)F ENDOR spectroscopy. AB - The binding of ethanol and 1,1,1-trifluoroethanol (TFE) to both the H(mv) and H(ox) forms of soluble methane monooxygenase (sMMO) in solution has been studied by Q-band (35 GHz) CW and pulsed ENDOR spectroscopy of (1)H, (2)H and (19)F nuclei of exogenous ligands. As part of this investigation we introduce (19)F, in this case from bound TFE, as a new probe for the binding of small molecules to a metalloenzyme active site. The H(mv) form was prepared in solution by chemical reduction of H(ox). For study of H(ox) itself, frozen solutions were subjected to gamma-irradiation in the frozen solution state at 77 K, which affords an EPR visible mixed-valent diiron center, denoted (H(ox))(mv), held in the geometry of the diiron(III) state. The (19)F and (2)H ENDOR spectra of bound TFE together with (1,2)H ENDOR spectra of bound ethanol indicate that the alcohols bind close to the Fe(II) ion of the mixed-valence cluster in H(mv) and in a bridging or semi bridging fashion to H(ox). DMSO does not affect the binding of either of the ethanols or of methanol to H(ox), nor of ethanol or methanol to H(mv). It does, however, displace TFE from the diiron site in H(mv). These results provide the first evidence that crystal structures of sMMO hydroxylase into which product alcohols were introduced by diffusion represent the structures in solution. PMID- 11890817 TI - Molecular structures and excited states of CpM(CO)(2) (Cp = eta(5)-C(5)H(5); M = Rh, Ir) and [Cl(2)Rh(CO)(2)](-). Theoretical evidence for a competitive charge transfer mechanism. AB - Molecular structures and excited states of CpM(CO)(2) (Cp = eta(5)-C(5)H(5); M = Rh, Ir) and [Cl(2)Rh(CO)(2)](-) complexes have been investigated using the B3LYP and the symmetry-adapted cluster (SAC)/SAC-configuration interaction (SAC-CI) theoretical methods. All the dicarbonyl complexes have singlet ground electronic states with large singlet-triplet separations. Thermal dissociations of CO from the parent dicarbonyls are energetically unfavorable. CO thermal dissociation is an activation process for [Cl(2)Rh(CO)(2)](-) while it is a repulsive potential for CpM(CO)(2). The natures of the main excited states of CpM(CO)(2) and [Cl(2)Rh(CO)(2)](-) are found to be quite different. For [Cl(2)Rh(CO)(2)](-), all the strong transitions are identified to be metal to ligand CO charge transfer (MLCT) excitations. A significant feature of the excited states of CpM(CO)(2) is that both MLCT excitation and a ligand Cp to metal and CO charge transfer excitation are strongly mixed in the higher energy states with the latter having the largest oscillator strength. A competitive charge transfer excited state has therefore been identified theoretically for CpRh(CO)(2) and CpIr(CO)(2). The wavelength dependence of the quantum efficiencies for the photoreactions of CpM(CO)(2) reported by Lees et al. can be explained by the existence of two different types of excited states. The origin of the low quantum efficiencies for the C-H/S-H bond activations of CpM(CO)(2) can be attributed to the smaller proportion of the MLCT excitation in the higher energy states. PMID- 11890818 TI - The nature of superacid electrophilic species in HF/SbF(5): a density functional theory study. AB - A density functional theory study at the B3LYP/6-31++G** + RECP(Sb) level of the HF/SbF(5) superacid system was carried out. The geometries of possible electrophilic species, such as H(2)F(+).Sb(2)F(11)(-) and H(3)F(2)(+).Sb(2)F(11)( ), were calculated and correspond with available experimental results. Calculations of different equilibrium reactions involving HF and SbF(5) allowed the relative concentration of the most energetically favorable species present in 1:1 HF/SbF(5) solutions to be estimated. These species are H(+).Sb(2)F(11)(-), H(2)F(+).Sb(2)F(11)(-), H(3)F(2)(+).Sb(2)F(11)(-), and H(4)F(3)(+).Sb(2)F(11)(-), which correspond to 36.9, 16.8, 36.9, and 9.4%, respectively. Calculations of the acid strength of the electrophilic species were also performed and indicated that, for the same anion, the acid strength increases with the solvation degree. The entropic term also plays a significant role in proton-transfer reactions in superacid systems. PMID- 11890820 TI - Mechanism of OH formation from ozonolysis of isoprene: a quantum-chemical study. AB - The formation and unimolecular reactions of primary ozonides and carbonyl oxides arising from the O(3)-initiated reactions of isoprene have been investigated using density functional theory and ab initio molecular orbital calculations. The activation energies of O(3) cycloaddition to the two double bonds of isoprene are found to be comparable (3.3-3.4 kcal mol(-1)), implying that the initial two O(3) addition pathways are nearly equally accessible. The reaction energies of O(3) addition to isoprene are between -47 and -48 kcal mol(-1). Cleavage of primary ozonides to form carbonyl oxides occurs with a barrier of 11-16 kcal mol(-1) above the ground state of the primary ozonide, and the decomposition energies range from -5 to -13 kcal mol(-1). OH formation is shown to occur primarily via decomposition of the carbonyl oxides with the syn-positioned methyl (alkyl) group, which is more favorable than isomerization to form dioxirane (by 1.1-3.3 kcal mol(-1)). Using the transition-state theory and master equation formalism, we determine an OH yield of 0.25 from prompt and thermal decomposition of the carbonyl oxides. PMID- 11890819 TI - Influence of d orbital occupation on the binding of metal ions to adenine. AB - Threshold collision-induced dissociation of M(+)(adenine) with xenon is studied using guided ion beam mass spectrometry. M(+) includes all 10 first-row transition metal ions: Sc(+), Ti(+), V(+), Cr(+), Mn(+), Fe(+), Co(+), Ni(+), Cu(+), and Zn(+). For the systems involving the late metal ions, Cr(+) through Cu(+), the primary product corresponds to endothermic loss of the intact adenine molecule, whereas for Zn(+), this process occurs but to form Zn + adenine(+). For the complexes to the early metal ions, Sc(+), Ti(+), and V(+), intact ligand loss competes with endothermic elimination of purine and of HCN to form MNH(+) and M(+)(C(4)H(4)N(4)), respectively, as the primary ionic products. For Sc(+), loss of ammonia is also a prominent process at low energies. Several minor channels corresponding to formation of M(+)(C(x)H(x)N(x)), x = 1-3, are also observed for these three systems at elevated energies. The energy-dependent collision-induced dissociation cross sections for M(+)(adenine), where M(+) = V(+) through Zn(+), are modeled to yield thresholds that are directly related to 0 and 298 K bond dissociation energies for M(+)-adenine after accounting for the effects of multiple ion-molecule collisions, kinetic and internal energy distributions of the reactants, and dissociation lifetimes. The measured bond energies are compared to those previously studied for simple nitrogen donor ligands, NH(3) and pyrimidine, and to results for alkali metal cations bound to adenine. Trends in these results and theoretical calculations on Cu(+)(adenine) suggest distinct differences in the binding site propensities of adenine to the alkali vs transition metal ions, a consequence of s-dsigma hybridization on the latter. PMID- 11890821 TI - Tunneling and sterically induced ring puckering in a substituted [8]annulene anion radical. AB - Electron paramagnetic resonance (EPR) studies have revealed that the steric interaction between the methyl hydrogens on a tert-butoxy substituent and the cyclooctatetraene (COT) ring system sterically induces a puckering of the eight membered ring in the anion radical of tert-butoxy-COT. The induced nonplanarity of the COT ring system causes a large attenuation of the EPR coupling constants. Since the C-D bond length is slightly shorter than is the C-H bond length, replacement of the tert-butyl group with a tert-butyl-d(9) group results in less steric interaction and measurably larger electron proton coupling constants. The oscillation between the two close to planar alternating bond length (ABL) D(2d) conformers of the COT moiety was found to be extremely rapid (k > 10(12) s(-1)) and quantum mechanical tunneling is proposed to be involved. PMID- 11890822 TI - Density functional calculations of electronic g-tensors for semiquinone radical anions. The role of hydrogen bonding and substituent effects. AB - A recently developed density functional approach has been used to carry out a systematic computational study of electronic g-tensors for a series of 1,4 semiquinone radical anions. Good agreement with high-field EPR data in frozen 2 propanol is achieved only after taking into account the significant reduction of g-tensor anisotropy caused by hydrogen bonding to solvent molecules. The comparison of various model systems for the first solvation shell suggests two hydrogen bonds from 2-propanol molecules to each of the carbonyl groups of the radical anions, and one additional hydrogen bond to each of the methoxy groups in ubiquinone systems. 2-Propanol makes stronger hydrogen bonds than water and thus influences g-tensor anisotropy more strongly. Substituent effects at the semiquinone are reproduced quantitatively by the calculations. The g-tensor anisotropy is influenced significantly by the conformations of methyl and methoxy substituents, with opposite contributions. Analyses and interpretations of the interrelations between structure, bonding, and spectroscopic data are provided. The relevance of the computational results for the EPR spectroscopy of semiquinone radical anions in photosynthetic reaction centers is discussed. PMID- 11890823 TI - De novo determination of protein backbone structure from residual dipolar couplings using Rosetta. AB - As genome-sequencing projects rapidly increase the database of protein sequences, the gap between known sequences and known structures continues to grow exponentially, increasing the demand to accelerate structure determination methods. Residual dipolar couplings (RDCs) are an attractive source of experimental restraints for NMR structure determination, particularly rapid, high throughput methods, because they yield both local and long-range orientational information and can be easily measured and assigned once the backbone resonances of a protein have been assigned. While very extensive RDC data sets have been used to determine the structure of ubiquitin, it is unclear to what extent such methods will generalize to larger proteins with less complete data sets. Here we incorporate experimental RDC restraints into Rosetta, an ab initio structure prediction method, and demonstrate that the combined algorithm provides a general method for de novo determination of a variety of protein folds from RDC data. Backbone structures for multiple proteins up to approximately 125 residues in length and spanning a range of topological complexities are rapidly and reproducibly generated using data sets that are insufficient in isolation to uniquely determine the protein fold de novo, although ambiguities and errors are observed for proteins with symmetry about an axis of the alignment tensor. The models generated are not high-resolution structures completely defined by experimental data but are sufficiently accurate to accelerate traditional high resolution NMR structure determination and provide structure-based functional insights. PMID- 11890824 TI - Determination of calpha chemical shift tensor orientation in peptides by dipolar modulated chemical shift recoupling NMR spectroscopy. AB - We present a new method for determining the orientation of chemical shift tensors in polycrystalline solids with site resolution and demonstrate its application to the determination of the Calpha chemical shift tensor orientation in a model peptide with beta-sheet torsion angles. The tensor orientation is obtained under magic angle spinning by modulating a recoupled chemical shift anisotropy (CSA) pattern with various dipolar couplings. These dipolar-modulated chemical shift patterns constitute the indirect dimension of a 2D spectrum and are resolved according to the isotropic chemical shifts of different sites in the direct dimension. These dipolar-modulated CSA spectra are equivalent to the projection of a 2D static separated-local-field spectrum onto its chemical shift dimension, except that its dipolar dimension is multiplied with a modulation function. Both (13)C-(1)H and (13)C-(15)N dipolar couplings can modulate the CSA spectra of the Calpha site in an amino acid and yield the relative orientations of the chemical shift principal axes to the C-H and C-N bonds. We demonstrate the C-H and C-N modulated CSA experiments on methylmalonic acid and N-tBoc-glycine, respectively. The MAS results agree well with the results of the 2D separated-local-field spectra, thus confirming the validity of this MAS dipolar-modulation approach. Using this technique, we measured the Val Calpha tensor orientation in N acetylvaline, which has beta-sheet torsion angles. The sigma(11) axis is oriented at 158 degrees (or 22 degrees) from the C-H bond, while the sigma(22) axis is tilted by 144 degrees (or 36 degrees) from the C-N bond. Both the orientations and the magnitude of this chemical shift tensor are in excellent agreement with quantum chemical calculations. PMID- 11890825 TI - Weak, improper, C-O...H-C hydrogen bonds in the dimethyl ether dimer. AB - The ground-state rotational spectrum of the dimethyl ether dimer, (DME)(2), has been studied by molecular beam Fourier transform microwave and free jet millimeter wave absorption spectroscopies. The molecular beam Fourier transform microwave spectra of the (DME-d(6))(2), (DME-(13)C)(2), (DME-d(6))...(DME), (DME (13)C)...(DME), and (DME)...(DME-(13)C) isotopomers have also been assigned. The rotational parameters have been interpreted in terms of a C(s) geometry with the two monomers bound by three weak C-H...O hydrogen bonds, each with an average interaction energy of about 1.9 kJ/mol. The experimental data combined with high level ab initio calculations show this kind of interaction to be improper, blue shifted hydrogen bonding, with an average shortening of the C-H bonds involved in the hydrogen bonding of 0.0014 A. The length of the C-H...O hydrogen bonds, r(O...H), is in the range 2.52-2.59 A. PMID- 11890826 TI - Theoretical determination of chromophores in the chromogenic effects of aromatic neurotoxicants. AB - We report the first computational study of the chromophores responsible for the chromogenic effects of aromatic neurotoxicants containing a 1,2-diacetyl moiety in their oxidation metabolites. A series of ab initio electronic structure calculations was performed on two representative aromatic compounds, 1,2 diacetylbenzene (1,2-DAB) and 1,2-diacetyl tetramethyl tetralin (1,2-DATT), the putative active metabolites of the neurotoxic aromatic hydrocarbon compounds 1,2 diethylbenzene (1,2-DEB) and acetyl ethyl tetramethyl tetralin (AETT), and on the products of their possible reactions with proteins that result in chromogenic effects. The electronic excitation energies determined by three different computational approaches were found to be consistent with each other. The calculated results are consistent with the conclusion/prediction that the chromogenic effects of 1,2-DAB (or 1,2-DEB) and 1,2-DATT (or AETT) could result from ninhydrin-like reactions, rather than the formation of pyrrole-like compounds. Our pK(a) calculations further indicate that the chromophore, i.e., the product of the ninhydrin-like reaction showing the blue color, is deprotonated in neutral aqueous solution. The corresponding protonated structure has a different color as it absorbs in the blue region of the visible spectrum, and its chromogenic contribution would be significant in solution at low pH. PMID- 11890827 TI - Theoretical studies of the cross-linking mechanisms between cytosine and tyrosine. AB - DNA-protein cross-linking is one of the many DNA lesions mediated by hydroxyl radicals, the most damaging among the reactive oxygen species in biological systems. Density functional theory methods are employed to investigate the complex reaction mechanisms of the formation of cytosine-tyrosine cross-links as observed in gamma-irradiated aqueous solutions of cytosine and tyrosine, as well as in gamma-irradiated nucleohistone. The majority of the radical addition mechanisms considered are found to have significant barriers and therefore to be thermodynamically unfavorable for the formation of the initial cross-linked product. Our calculated reaction potential energy surfaces suggest that a feasible complete mechanism consists of radical combination forming the initial cross-linked product, a hydrogen shuffle within the initial cross-linked product, and an acid-catalyzed dehydration reaction. Water and hydrogen-bonding interactions are suggested to play a key role in catalyzing the hydrogen-transfer step of the reaction. PMID- 11890828 TI - Relativistic spin-orbit coupling effects on secondary isotope shifts of (13)C nuclear shielding in CX(2) (X = O, S, Se, Te). AB - Rovibrational corrections, temperature dependence, and secondary isotope shifts of the (13)C nuclear shielding in CX(2) (X = O, S, Se, Te) are calculated taking into account the relativistic spin-orbit (SO) interaction. The SO effect is considered for the first time for the secondary isotope shifts. The nuclear shielding hypersurface in terms of nuclear displacements is calculated by using a density-functional theory method. Ab initio multiconfiguration self-consistent field calculations are done at the equilibrium geometry for comparison. (13)C NMR measurements are carried out for CS(2). The calculated results are compared with both present and earlier experimental data on CO(2), CS(2), and CSe(2). The heavy atom SO effects on the rovibrational corrections of (13)C shielding are shown to be significant. For CSe(2) and CTe(2), reliable prediction of secondary isotope effects and their temperature dependence requires the inclusion of the SO corrections. In particular, earlier discrepancies of theory and experiment for CSe(2) are fully resolved by taking the SO interactions into account. PMID- 11890829 TI - Adsorption geometry of 4-picoline chemisorbed on the Cu(110) surface: a study of forces controlling molecular self-assembly. AB - The adsorption of 4-picoline (4-methylpyridine) on the Cu(110) surface has been studied with time-of-flight electron stimulated desorption ion angular distribution (TOF-ESDIAD) and other methods. Using deuterium labeling in the methyl group and hydrogen labeling on the aromatic ring, it has been possible to separately monitor by TOF-ESDIAD the C-D bond directions and the C-H bond directions in the adsorbed molecule. These triangulation measurements have led to a detailed understanding of the conformation of the adsorbed molecule relative to the Cu(110) crystal lattice, allowing one to witness changes in the molecular conformation as adsorbate-adsorbate interactional effects take place for increasing coverages. At low coverages, the molecule adsorbs by the N atom at an atop Cu site with the aromatic ring parallel to the <001> azimuth and with the molecular axis inclined 33 (+/- 5) degrees along the <001> azimuth. As rows of 4 picoline molecules form long range ordered chain structures oriented along the <112> azimuth, the aromatic ring twists 29 degrees about the inclined molecular axis as a result of forces between the adsorbate molecules. The initial tilting of the molecular axis at low coverage is likely due to the interaction of the positive-outward dipole with its image in the substrate. The ring twist may result from dipoleminus signdipole forces between the adsorbate molecules in the rows formed tending to form nested parallel pyridine rings. These studies are the first to apply the TOF-ESDIAD method for the measurement of the direction of chemical bonds at more than one molecular location within an adsorbed molecule and the new method is named electron stimulated desorption-molecular triangulation (ESD-MT). The results obtained give information of importance in understanding the factors which control conformational effects during the molecular self-assembly of complex adsorbed molecules on surfaces. PMID- 11890830 TI - Reaction of phenyl radicals with propyne. AB - The potential energy surface (PES) for the phenyl + propyne reaction, which might contribute to the growth of polycyclic aromatic hydrocarbons (PAHs) under a wide variety of reaction conditions, is described. The PES was characterized at the B3LYP-DFT/6-31G(d) and B3LYP-DFT/6-311+G(d,p) levels of theory. The energies of the entrance transition states, a direct hydrogen-transfer channel and two addition reactions leading to chemically activated C(9)H(9) intermediates, were also evaluated at the QCISD(T)/ 6-311G(d,p) and CCSD(T)/6-311G(d,p) levels of theory. An extensive set of unimolecular reactions was examined for these activated C(9)H(9)(dagger) intermediates, comprising 70 equilibrium structures and over 150 transition states, and product formation channels leading to substituted acetylenes and allenes such as PhCCH, PhCCCH(3), and PhCHCCH(2) were identified. The lowest energy pathway leads to indene, a prototype PAH molecule containing a five-membered ring. The title reaction thus is an example of possible direct formation of a PAH containing a five-membered ring, necessary to explain formation of nonplanar PAH structures, from an aromatic radical unit and an unsaturated hydrocarbon bearing an odd number of carbons. Extensive Supporting Information is available. PMID- 11890831 TI - Formation of a 1-bicyclo[1.1.1]pentyl anion and an experimental determination of the acidity and C-H bond dissociation energy of 3-tert butylbicyclo[1.1.1]pentane. AB - Decarboxylation of 1-bicyclo[1.1.1]pentanecarboxylate anion does not afford 1 bicyclo[1.1.1]pentyl anion as previously assumed. Instead, a ring-opening isomerization which ultimately leads to 1,4-pentadien-2-yl anion takes place. A 1 bicyclo[1.1.1]pentyl anion was prepared nevertheless via the fluoride-induced desilylation of 1-tert-butyl-3-(trimethylsilyl)bicyclo[1.1.1]pentane. The electron affinity of 3-tert-butyl-1-bicyclo[1.1.1]pentyl radical (14.8 plus minus 3.2 kcal/mol) was measured by bracketing, and the acidity of 1-tert butylbicyclo[1.1.1]pentane (408.5 +/- 0.9) was determined by the DePuy kinetic method. These values are well-reproduced by G2 and G3 calculations and can be combined in a thermodynamic cycle to provide a bridgehead C-H bond dissociation energy (BDE) of 109.7 +/- 3.3 kcal/mol for 1-tert-butylbicyclo[1.1.1]pentane. This bond energy is the strongest tertiary C-H bond to be measured, is much larger than the corresponding bond in isobutane (96.5 +/- 0.4 kcal/mol), and is more typical of an alkene or aromatic compound. The large BDE can be explained in terms of hybridization. PMID- 11890832 TI - Field-dependent electrode-chemisorbate bonding: sensitivity of vibrational stark effect and binding energetics to nature of surface coordination. AB - Illustrative quantum-chemical calculations for selected atomic and molecular chemisorbates on Pt(111) (modeled as a finite cluster) are undertaken as a function of external field, F, by using Density Functional Theory (DFT) with the aim of ascertaining the sensitivity of the field-dependent metal-adsorbate binding energetics and vibrational frequencies (i.e., the vibrational Stark effect) to the nature of the surface coordination in electrochemical systems. The adsorbates selected--Cl, I, O, N, Na, NH(3), and CO--include chemically important examples featuring both electron-withdrawing and -donating characteristics. The direction of metal-adsorbate charge polarization, characterized by the static dipole moment, mu(S), determines the binding energy-field (E(b-F) slopes, while the corresponding Stark-tuning behavior is controlled primarily by the dynamic dipole moment, mu(D). Significantly, analysis of the F-dependent sensitivity of mu(S) and mu(D) leads to a general adsorbate classification. For electronegative adsorbates, such as O and Cl, both mu(S) and mu(D) are negative, the opposite being the case for electropositive adsorbates. However, for systems forming dative-covalent rather than ionic bonds, as exemplified here by NH(3) and CO, mu(S) and mu(D) have opposite signs. The latter behavior, including electron donating and -withdrawing categories, arises from diminishing metal-chemisorbate orbital overlap, and hence the extent of charge polarization, as the bond is stretched. A clear-cut distinction between these different types of surface bonding is therefore obtainable by combining vibrational Stark-tuning and E(b)-F slopes, as extracted from experimental data and/or DFT calculations. The former behavior is illustrated by means of potential-dependent Raman spectral data obtained in our laboratory. PMID- 11890833 TI - Searching for the second oxidant in the catalytic cycle of cytochrome P450: a theoretical investigation of the iron(III)-hydroperoxo species and its epoxidation pathways. AB - Iron(III)-hydroperoxo, [Por(CysS)Fe(III)-OOH](-), a key species in the catalytic cycle of cytochrome P450, was recently identified by EPR/ENDOR spectroscopies (Davydov, R.; Makris, T. M.; Kofman, V.; Werst, D. E.; Sligar, S. G.; Hoffman, B. M. J. Am. Chem. Soc. 2001, 123, 1403-1415). It constitutes the last station of the preparative steps of the enzyme before oxidation of an organic compound and is implicated as the second oxidant capable of olefin epoxidation (Vaz, A. D. N.; McGinnity, D. F.; Coon, M. J. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 3555-3560), in addition to the penultimate active species, Compound I (Groves, J. T.; Han, Y. Z. In Cytochrome P450: Structure, Mechanism and Biochemistry, 2nd ed.; Ortiz de Montellano, P. R., Ed.; Plenum Press: New York, 1995; pp 3-48). In response, we present a density functional study of a model species and its ethylene epoxidation pathways. The study characterizes a variety of properties of iron(III)-hydroperoxo, such as the O-O bonding, the Fe-S bonding, Fe-O and Fe-S stretching frequencies, its electron attachment, and ionization energies. Wherever possible these properties are compared with those of Compound I. The proton affinities for protonation on the proximal and distal oxygen atoms of iron(III)-hydroperoxo, and the effect of the thiolate ligand thereof, are determined. In accordance with previous results (Harris, D. L.; Loew, G. H. J. Am. Chem. Soc. 1998, 120, 8941-8948), iron(III)-hydroperoxo is a strong base (as compared with water), and its distal protonation leads to a barrier-free formation of Compound I. The origins of this barrier-free process are discussed using a valence bond approach. It is shown that the presence of the thiolate is essential for this process, in line with the "push effect" deduced by experimentalists (Sono, M.; Roach, M. P.; Coulter, E. D.; Dawson, J. H. Chem. Rev. 1996, 96, 2841-2887). Finally, four epoxidation pathways of iron(III) hydroxperoxo are located, in which the species transfers oxygen to ethylene either from the proximal or from the distal sites, in both concerted and stepwise manners. The barriers for the four mechanisms are 37-53 kcal/mol, in comparison with 14 kcal/mol for epoxidation by Compound I. It is therefore concluded that iron(III)-hydroperoxo, as such, cannot be a second oxidant, in line with its significant basicity and poor electron-accepting capability. Possible versions of a second oxidant are discussed. PMID- 11890834 TI - Quantitative cavities and reactivity in stages of crystal lattices: mechanistic and exploratory organic photochemistry. AB - In continuing our research on solid-state organic photochemistry, we have been investigating the phenomenon of reactivity in stages. In this study we present new examples where the photochemical reactivity changes discontinuously at some point in the conversion. In these instances, the reaction course of the solid differs from that in solution. One example is the reaction of 2-methyl-4,4 diphenylcyclohexenone, where an unusual reaction course was encountered in the solid state; and, of two possible mechanisms, one was established by isotopic labeling. A second case is that of 4,5,5-triphenylcyclohexenone. The solid-state reaction of this enone was found to give a new photochemical transformation, the Type C rearrangement, a process that involves a delta to alpha aryl migration. In the case of 3-tert-butyl-5,5-diphenylcyclohexenone the Type C rearrangement occurred even in solution. The stage behavior was investigated using X-ray analysis and Quantum Mechanics/Molecular Mechanics computations. This permitted us to determine the sources and details of the stage phenomenon. The analysis revealed how a product molecule as a neighbor affects reactivity. The computations were employed to follow the course of a solid-state reaction from reactant through the succeeding stages. Additionally, the Delta-Density Analysis was utilized to ascertain the electronic nature of molecular changes. Besides product composition changing with extent of conversion, the reaction quantum yield was found to change as one stage gave way to a succeeding one. PMID- 11890835 TI - Nucleophilic or electrophilic phosphinidene complexes ML(n)=PH; what makes the difference? AB - Density functional studies, based on the local density approximation including nonlocal corrections for correlation and exchange self-consistently, have been carried out for the equilibrium structures of the phosphinidene transition metal complexes ML(n)=PH, with M = Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, W, Fe, Ru, Os, Co, Rh, Ir and L = CO, PH(3), Cp. The chemical reactivity of the transition metal stabilized phosphinidene P-R is influenced by its spectator ligands L. Ligands with strong sigma-donor capabilities on the metal increase the electron density on the phosphorus atom, raise the pi-orbital energy, and enhance its nucleophilicity. Spectator ligands with strong pi-acceptor capabilities lower the charge concentration on P and stabilize the pi-orbital, which results in a higher affinity for electron-rich species. The ML(n)=PH bond is investigated using a bond energy analysis in terms of electrostatic interaction, Pauli repulsion, and orbital interaction. A symmetry decomposition scheme affords a quantitative estimate of the sigma- and pi-bond strengths. It is shown that the investigated phosphinidenes are strong pi-acceptors and even stronger sigma-donors. The metal phosphinidene interaction increases on going from the first to the second- and third-row transition metals. PMID- 11890836 TI - Vinyl-vinyl coupling on late transition metals through C-C reductive elimination mechanism. A computational study. AB - A detailed density functional study was performed for the vinyl-vinyl reductive elimination reaction from bis-sigma-vinyl complexes [M(CH=CH(2))(2)X(n)]. It was shown that the activity of these complexes decreases in the following order: Pd(IV), Pd(II) > Pt(IV), Pt(II), Rh(III) > Ir(III), Ru(II), Os(II). The effects of different ligands X were studied for both platinum and palladium complexes, which showed that activation barriers for C-C bond formation reaction decrease in the following order: X = Cl > Br, NH(3) > I > PH(3). Steric effects induced either by the ligands X or by substituents on the vinyl group were also examined. In addition, the major factors responsible for stereoselectivity control on the final product formation stage and possible involvement of asymmetric coupling pathways are reported. In all cases DeltaE, DeltaH, DeltaG, and DeltaG(aq) energy surfaces were calculated and analyzed. The solvent effect calculation shows that in a polar medium halogen complexes may undergo a reductive elimination reaction almost as easily as compounds with phosphine ligands. PMID- 11890837 TI - A modern approach to posterior circulation ischemic stroke. PMID- 11890838 TI - Disorders of cortical development and epilepsy. AB - There has been an impressive increase in our ability to identify and categorize patients with cortical development lesions over the past decade. The clinical features associated with disorders of cortical development (DCD) have been described, and epilepsy has been shown to be a frequent symptom. In this review, we categorize DCD based on their structure and discuss their underlying causes and clinical features. Just as the cause of each type of disorder is thought to be unique, each disorder also has distinct types of seizures, treatment strategies, and electroencephalographic features. Studies in human tissue and animal models of DCD have begun to shed light on why DCD are associated with epilepsy. Aberrant synaptic connections within the dysplastic tissue and between the dysplastic tissue and more normal-appearing adjacent tissue form an abnormal, hyperexcitable network that increases seizure susceptibility. In the future, strategies for blocking formation of the aberrant networks may prevent the development of epilepsy. PMID- 11890839 TI - Outcome at 30 days in the New England Medical Center Posterior Circulation Registry. AB - BACKGROUND: Vertebrobasilar disease is generally considered a condition with a poor prognosis because of high rates of mortality and severe disability. OBJECTIVE: To compare the outcomes of 407 patients entered in the New England Medical Center Posterior Circulation Registry with the reported results of other studies. RESULTS: In contrast, among 407 patients prospectively and consecutively studied in the New England Medical Center Posterior Circulation Registry, we found a low mortality rate at 30 days after onset (3.6%) and relatively low rates of major disability (18% using a Modified Rankin Disability Scale score). Thirty days after stroke, 28% of the patients had no disability and 51% had only a minor disability. Stroke location, stroke mechanism, and arteries involved predicted outcome. Basilar artery involvement, embolic stroke mechanism, and multiple posterior circulation intracranial territory involvement correlated with poor outcome. Patients with lesions in the basilar artery were 5 times more likely to have a poor outcome independent of other factors. Lesions in the middle and distal territories were each associated with a poor outcome in one third of the patients. CONCLUSION: In contrast with previous reports, we found that vertebrobasilar occlusive disease consists of a variety of different stroke mechanisms and vascular lesions, many with a good prognosis. PMID- 11890840 TI - Serum lipoprotein levels, statin use, and cognitive function in older women. AB - BACKGROUND: Few strategies are available for the prevention of cognitive impairment in elderly persons. Serum lipoprotein levels may be important predictors of cognitive function, and drugs that lower cholesterol may be effective for the prevention of cognitive impairment. OBJECTIVE: To determine whether serum lipoprotein levels, the 4-year change in serum lipoprotein levels, and the use of statin drugs are associated with cognition in older women without dementia. DESIGN, SETTING, AND PARTICIPANTS: An observational study of 1037 postmenopausal women with coronary heart disease enrolled in the Heart and Estrogen/progestin Replacement Study (participants at 10 of 20 centers). MAIN OUTCOME MEASURE: The Modified Mini-Mental State Examination was administered at the end of the study after 4 years of follow-up. Women whose score was less than 84 points (>1.5 SDs below the mean) were classified as having cognitive impairment. Lipoprotein levels (total, high-density lipoprotein, and low-density lipoprotein [LDL] cholesterol and triglycerides) were measured at baseline and at the end of the study; statin use was documented at each visit. RESULTS: Compared with women in the lower quartiles, women in the highest LDL cholesterol quartile at cognitive testing had worse mean plus minus SD Modified Mini-Mental State Examination scores (93.7 plus minus 6.0 vs 91.9 plus minus 7.6; P =.002) and an increased likelihood of cognitive impairment (adjusted odds ratio, 1.76; 95% confidence interval, 1.04-2.97). A reduction in the LDL cholesterol level during the 4 years tended to be associated with a lower odds of impairment (adjusted odds ratio, 0.61; 95% confidence interval, 0.36-1.03) compared with women whose levels increased. Higher total and LDL cholesterol levels, corrected for lipoprotein(a) levels, were also associated with a worse Modified Mini-Mental State Examination score and a higher likelihood of impairment, whereas high density lipoprotein cholesterol and triglyceride levels were not associated with cognition. Compared with nonusers, statin users had higher mean plus minus SD Modified Mini-Mental State Examination scores (92.7 plus minus 7.1 vs 93.7 plus minus 6.1; P =.02) and a trend for a lower likelihood of cognitive impairment (odds ratio, 0.67; 95% confidence interval, 0.42-1.05), findings that seemed to be independent of lipid levels. CONCLUSIONS: High LDL and total cholesterol levels are associated with cognitive impairment, and lowering these lipoprotein levels may be a strategy for preventing impairment. The association between statin use and better cognitive function in women without dementia requires further study. PMID- 11890841 TI - Endogenous estradiol in elderly individuals: cognitive and noncognitive associations. AB - OBJECTIVE: To investigate an association between endogenous estradiol (E(2)) levels and cognition and behavior in elderly individuals. PATIENTS: We studied 135 community-based men and women aged 52 to 85 years in urban Bangkok, Thailand; 72 had dementia and 63 did not. MATERIALS AND METHODS: Dementia was diagnosed using Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria after appropriate investigations. Blood samples for assay were collected in the morning after 6 hours of fasting. Levels of E(2) were measured by radioimmunoassay (double antibody technique). The Thai version of the Mini-Mental State Examination was used to assess cognition; the Neuropsychiatric Inventory was used to assess neuropsychiatric symptoms; and the Functional Assessment Questionnaire was used to assess instrumental activities of daily living. RESULTS: There was no correlation between age and level of E(2) in either men or women. Individuals with lower estrogen levels had more behavioral disturbances (men: r = -0.467, n = 45; P =.001; women: r = -0.384, n = 90; P<.001) and worse cognition (men: r = 0.316, n = 45; P =.03; women: r = 0.243, n = 90; P =.02) and function (men: r = -0.417, n = 45; P =.004; women: r = -0.437, n = 90; P<.001). The threshold level of endogenous E(2) in elderly individuals for the risk of developing dementia was less than 15 pg/mL (<55 pmol/L) in men and less than 1 pg/mL (<4 pmol/L) in women. CONCLUSION: Lower E(2) levels are correlated with poor cognitive, behavioral, and functional status in older individuals. PMID- 11890842 TI - Migration of multiple sclerosis lymphocytes through brain endothelium. AB - CONTEXT: T-lymphocyte migration through the blood-brain barrier is a central event in the process of lesion formation in multiple sclerosis (MS). OBJECTIVES: To assess the ability of lymphocytes derived from the peripheral blood of patients with clinically active and inactive MS to migrate across an artificial model of the blood-brain barrier and to elucidate the molecular mechanisms involved in such a process. DESIGN: We developed an in vitro model of lymphocyte migration using a Boyden chamber coated with a monolayer of human brain microvascular endothelial cells. RESULTS: The rates of migration of lymphocytes obtained from patients with acutely relapsing and active secondary progressive MS was significantly increased compared with those obtained from healthy controls and patients with inactive secondary progressive disease. Ribonuclease protection assays and enzyme-linked immunosorbent assays indicated that monocyte chemoattractant protein 1 and interleukin 8 were the major chemokines produced by brain endothelial cells grown under the culture conditions used for the migration assays. The rate of migration of the MS lymphocytes could be inhibited by 60% with an antimonocyte chemoattractant protein 1 monoclonal antibody, indicating a functional role for this chemokine in the migration process. In agreement with previous reports, we found that the tissue inhibitor of metalloproteinase 1, a matrix metalloproteinase inhibitor, also reduced migration of MS lymphocytes by 50%. CONCLUSIONS: The results demonstrate an increased migration rate of MS T lymphocytes across the brain endothelium barrier and that such migration is dependent on chemokine monocyte chemoattractant protein 1 and on matrix metalloproteinases. PMID- 11890843 TI - Magnetic resonance imaging in basilar artery occlusion. AB - CONTEXT: Acute basilar artery occlusion has particularly high mortality and morbidity. OBJECTIVE: To determine the potential utility of advanced magnetic resonance imaging (MRI) methods, including diffusion-weighted imaging, for the early management of patients with basilar artery thrombosis. DESIGN: Case series. SETTING: Institute of Neuroradiology and Department of Neurology, Johann Wolfgang Goethe University, Frankfurt, Germany. PATIENTS: In 4 patients with occlusion of the basilar artery, MRI was performed, including T2-weighted and diffusion weighted imaging (DWI) sequences and magnetic resonance angiography (MRA) in the short-term phase (<12 hours). Three patients underwent intra-arterial thrombolysis. Clinical outcome was obtained 10 days after symptom onset. RESULTS: The MRA was performed 3.5 to 11.5 hours after symptom onset and showed basilar artery occlusion in all cases. The DWI revealed different patterns of ischemic lesions. In 2 patients, no or only small lesions could be identified; the remaining showed multiple and large lesions within the posterior circulation territory. Initial clinical status was severely impaired in all cases (Rankin scale score, 4-5). Thrombolysis was initiated in 3 patients, leading to successful recanalization in 2. Clinical outcome was favorable in the 2 patients with small DWI lesions and successful reperfusion (Rankin scale score, 2), whereas it was worse in those with large DWI lesions and persisting occlusion (death, persisting coma). CONCLUSIONS: In critically ill patients with acute basilar occlusion, the extent of DWI lesion involvement can be highly variable. Small DWI lesions seem to be associated with a favorable outcome if reperfusion is achieved with thrombolysis. This could potentially be the case independent of time from symptom onset. PMID- 11890845 TI - Transcranial magnetic stimulation of the cerebellum in essential tremor: a controlled study. AB - BACKGROUND: Growing evidence implicates an overactivity of the cerebellum in the pathophysiology of essential tremor. In a small series of patients, we explored the acute effects and therapeutic possibilities of low-frequency repetitive transcranial magnetic stimulation (rTMS) of the cerebellum in patients with essential tremor in a double-blind, crossover, placebo-controlled design. METHODS: Ten patients with essential tremor underwent an active and a sham rTMS session, at a 1-week interval. The rTMS was performed with a focal double 70-mm butterfly coil (maximum peak field of 2.2 T) applied 2 cm below the inion. Each session consisted of 30 trains of 10-second duration separated by 30-second pauses, at 100% of the maximum output intensity and at 1-Hz frequency. Major evaluation outcomes were the score on the Tremor Clinical Rating Scale and accelerometric recordings obtained before (-5 minutes), immediately after (+5 minutes), and 1 hour after (+60 minutes) each rTMS session. Both clinical and accelerometric measurements were obtained by a blinded neurologist. RESULTS: On the +5-minute assessment, active rTMS produced a notable tremor improvement compared with sham rTMS, as evidenced by a significant reduction in scores on the clinical rating scale and accelerometric values. At +60 minutes, no clinical or accelerometric benefit was evidenced. No adverse effects of rTMS were observed. CONCLUSIONS: This exploratory study of the potential therapeutic properties of rTMS on essential tremor showed an acute antitremor effect. Further investigation in search of a more lasting benefit is warranted. PMID- 11890844 TI - Association of a null mutation in the CNTF gene with early onset of multiple sclerosis. AB - BACKGROUND: Immune-mediated demyelination and axonal damage lead to early functional impairment in multiple sclerosis (MS). Ciliary neurotrophic factor (CNTF) is a potent survival factor for neurons and oligodendrocytes and may be relevant in reducing tissue destruction during inflammatory attacks. SUBJECTS AND METHODS: We screened 288 unselected patients with multiple sclerosis (MS) (mean age, 40.2 +/- 10.2 years; range, 18-71 years) for a previously described homozygous null mutation within the CNTF gene leading to a truncated, biologically inactive protein. The G-to-A CNTF null mutation at position -6 of the second exon was identified by a HaeIII polymorphism of the polymerase chain reaction-amplified genomic DNA. RESULTS: The homozygous CNTF null mutation (CNTF /-) was found in 7 (2.4%) of the 288 randomly selected patients with MS. Patients with the CNTF -/- genotype had a significantly earlier onset of disease (17 vs 27 years; Mann-Whitney test, P =.007) with predominant motor symptoms. CONCLUSIONS: These results suggest that CNTF contributes to time and site of early clinical manifestation. The frequency of patients with MS with a homozygous CNTF null mutation in this population was not higher than in control groups, indicating that the CNTF null mutation is not a risk factor for development of MS. PMID- 11890846 TI - Botulinum toxin A treatment for primary hemifacial spasm: a 10-year multicenter study. AB - BACKGROUND: Botulinum toxin A (BTX) is the currently preferred symptomatic treatment for primary hemifacial spasm (HFS), but its long-term efficacy and safety are not known. OBJECTIVE: To assess the long-term effectiveness and safety of BTX in the treatment of primary HFS. DESIGN: Retrospective review of medical records of the 1st and 10th years of treatment. SETTING: Outpatient clinics of 4 Italian university centers in the Italian Movement Disorders Study Group. PARTICIPANTS: A series of 65 patients with primary HFS who had received BTX injections regularly for at least 10 years. MAIN OUTCOME MEASURES: Mean duration of improvement and quality of the effect induced by the preceding treatment (measured using a patient self-evaluation scale) and occurrence and duration of adverse effects in the 1st and 10th years of treatment. RESULTS: Using a mean BTX dose per treatment session similar to that used by others, we obtained a 95% response rate and an overall mean duration of improvement of 12.6 weeks during year 1. The effectiveness of BTX in relieving the symptoms of primary HFS, as measured by the response rate and average duration of improvement, remained unchanged in the 1st and 10th years. Patients needed statistically similar BTX doses in the 1st and 10th years. The rate of local adverse effects (including upper lid ptosis, facial weakness, and diplopia) diminished significantly in the 10th year of treatment. CONCLUSION: Treatment with BTX effectively induces sustained relief from symptoms of HFS in the long term, with only minimal and transient adverse reactions. PMID- 11890847 TI - Exploring the relationship between Parkinson disease and restless legs syndrome. AB - BACKGROUND: Restless legs syndrome (RLS) and Parkinson disease (PD) are common neurological conditions that respond to dopaminergic therapy. To our knowledge, the relationship between the two has not been thoroughly explored. METHODS: We consecutively queried 303 patients with PD seen in our clinic for the presence of RLS symptoms, and evaluated their condition with the Epworth Sleepiness Scale and other demographic and sleep measures. We then looked for predictors of RLS in these patients with PD. We also compared a larger group of patients with PD/RLS with a group of patients with RLS alone. RESULTS: Of 303 patients with PD, 63 (20.8%) had symptoms of RLS. Neither PD patient demographics nor PD treatments could reliably predict the development of RLS symptoms; however, lower serum ferritin levels were associated with RLS symptoms in our patients with PD (P =.01). In 54 (68%) of the 79 total patients with PD/RLS (including additional patients with PD/RLS seen in the clinic) with reliable age-at-onset data, the PD symptoms preceded the RLS symptoms (chi(2) test, P<.001). Compared with patients with idiopathic RLS (N = 146), patients with PD/RLS (N = 109) were older at RLS onset (P<.001), were less likely to have a family history of RLS (P<.001), and had lower serum ferritin levels (P =.01). CONCLUSIONS: Symptoms of RLS are common in patients with PD; however, except in patients with a family history of RLS, they seem to reflect a secondary phenomenon, perhaps in relation with lower ferritin levels. There is no evidence that RLS symptoms early in life predispose to the subsequent development of PD. PMID- 11890848 TI - Gradient echo magnetic resonance imaging in the prediction of hemorrhagic vs ischemic stroke: a need for the consideration of the extent of leukoariosis. AB - BACKGROUND: Multifocal signal loss lesion (MSLL) on gradient echo magnetic resonance imaging (GE-MRI) may reflect bleeding-prone microangiopathy. However, MSLLs are also known to be associated with leukoariosis; leukoariosis is commonly associated with occlusive-type vascular lesions. OBJECTIVE: To determine whether MSLL on GE-MRI is significantly associated with the type of stroke--intracerebral hemorrhagic (ICH) stroke more often than an ischemic stroke (infarction)- regardless of the extent of leukoariosis. PATIENTS AND METHODS: We studied 91 patients who had an acute stroke and were admitted to the Department of Neurology, Seoul National University Hospital, Seoul, South Korea, from March 1, 1997, to July 31, 1998. These patients underwent both conventional MRI and GE MRI. The GE-MRI was used to count MSLLs. We also counted lacunae and classified leukoariosis (none or mild and advanced). Multiple logistic regression analysis was used to test for MSLL-leukoariosis interaction association with the type of stroke (ICH over infarction) and to evaluate the relative contribution of an MSLL -adjusted for age, sex, and lacunae--in discriminating the type of stroke. RESULTS: The association between MSLL and ICH statistically significantly differed by leukoariosis (P =.003 for MSLL-leukoariosis interaction term). The MSLL count on GE-MRI was significantly associated with the type of stroke (ICH over infarction; odds ratio, 2.46; 95% confidence interval, 1.38-4.39) when leukoariosis was classified as none or mild. When leukoariosis was classified as advanced, there was a decrease in the odds ratio of MSLL to 0.99 (95% confidence interval, 0.94-1.04). CONCLUSIONS: Our findings indicate that MSLL on GE-MRI is a predictor of ICH vs infarction in patients with no or mild leukoariosis, but not in patients with advanced leukoariosis. Therefore, in the evaluation of GE-MRI for a bleeding-prone microangiopathy, the extent of leukoariosis should be considered. PMID- 11890849 TI - Reduced aquaporin 4 expression in the muscle plasma membrane of patients with Duchenne muscular dystrophy. AB - BACKGROUND: In Duchenne muscular dystrophy (DMD), previous freeze-fracture electron microscopic studies demonstrated that muscle plasma membrane contained markedly decreased numbers of orthogonal arrays. Recent investigations showed that orthogonal arrays were composed of aquaporin 4 (AQP4) molecules, a member of the water channel protein family. OBJECTIVES: To study whether the immunostainability of anti-AQP4 antibody is reduced in muscles of patients with DMD and whether, if it is reduced, the problem is at the genomic DNA, messenger RNA (mRNA), or posttranscriptional level. PATIENTS AND METHODS: We analyzed the muscle and blood samples from 6 boys with DMD, 6 normal control subjects, and 12 patients with neuromuscular diseases at the protein, genomic DNA, and mRNA levels. At the protein level, immunohistochemical staining and immunoblot analysis were performed. At the genomic DNA and mRNA levels, the polymerase chain reaction and reverse transcription polymerase chain reaction, respectively, were used to screen for mutations in the AQP4 gene. RESULTS: At the protein level, immunohistochemical staining of our originally generated rabbit anti-AQP4 antibody in DMD muscles was markedly reduced. Most of the DMD myofibers showed negative staining with sporadic partially positive fibers at their myofiber surface, whereas the control muscles displayed continuous myofiber surface staining. Immunoblot analysis showed that the content of AQP4 in DMD muscles was remarkably decreased. Amplification of leukocyte genomic DNA by polymerase chain reaction showed that the patients with DMD had genomic DNA of the AQP4 molecule. Quantitative reverse transcription polymerase chain reaction demonstrated that DMD skeletal muscles contained markedly decreased AQP4 mRNA compared with controls. CONCLUSION: The reduction in AQP4 in DMD muscles results from decreased levels of AQP4 mRNA in DMD myofibers. PMID- 11890850 TI - The neurological masquerade of intravascular lymphomatosis. AB - BACKGROUND: Intravascular lymphomatosis (IVL) is an uncommon systemic disease characterized by occlusion of small vessels by malignant lymphomatous cells. Central nervous system involvement usually presents as subacute encephalopathy, dementia, seizures, or multifocal cerebrovascular events. OBJECTIVE: To increase awareness about IVL, an uncommon cause of neurological disease. DESIGN: This is a retrospective case series of 8 pathologically proved cases of IVL with neurological disease. Patients were part of a pathological series collected between April 1962 and October 1998 at Indiana University School of Medicine and the Armed Forces Institute of Pathology, Washington, DC. SETTING: Neurological and neuropathological examinations were performed at tertiary referral hospitals. PATIENTS: Eleven patients were diagnosed pathologically as having IVL, but 3 were not included in this evaluation because of a lack of appropriate clinical information. Of the final sample (n = 8), there were 4 men and 4 women (mean +/- SD age, 62.9 +/- 9.9 years). RESULTS: All 8 patients had focal neurological deficits, 7 had encephalopathy or dementia, 5 had epileptic seizures, and 2 had myelopathy. Death occurred at a mean of 7.7 months (range, 1-24 months) after the onset of symptoms. All patients had elevated cerebrospinal fluid protein levels, 4 had pleocytosis, and 2 had an elevated IgG level in their cerebrospinal fluid. Of the 4 patients who underwent a brain biopsy, 1 was diagnosed as having IVL before death. CONCLUSIONS: Intravascular lymphomatosis is an uncommon disease with a myriad of potential neurological manifestations. Diagnosis requires a high index of suspicion and a pathological examination. If diagnosed early, aggressive chemotherapy is potentially curative, although the overall prognosis remains dismal. PMID- 11890852 TI - Endovascular closure of a patent foramen ovale in the fat embolism syndrome: changes in the embolic patterns as detected by transcranial Doppler. AB - BACKGROUND: The posttraumatic fat embolism syndrome (FES) is characterized by petechiae and pulmonary and cerebral dysfunction. A patent foramen ovale (PFO) could worsen the prognosis of FES by allowing larger emboli to reach the systemic circulation. Transcranial Doppler ultrasonography can be used to diagnose and monitor cerebral microembolism in FES. OBJECTIVE: To describe a case of successful percutaneous closure of PFO in a patient with posttraumatic FES with excellent clinical outcome. PATIENT AND METHODS: A 17-year-old girl presented with a posttraumatic long-bone fracture complicated by typical severe FES. Transcranial Doppler disclosed multiple microembolic signals over both middle cerebral and basilar arteries. A large PFO was diagnosed by transesophageal echocardiogram. A brain magnetic resonance image with diffusion-weighted sequences showed multiple bilateral areas of abnormal diffusion in watershed territories. Percutaneous PFO closure with a buttoned device was successfully performed. RESULTS: Closure of PFO was associated with marked reduction in the number and intensity of microembolic signals. Subsequent surgical repair of the fracture with the patient under transcranial Doppler monitoring was uneventful. There was excellent correlation between clinical course and microembolic signal load by transcranial Doppler. CONCLUSIONS: Cerebral fat embolism after long-bone fractures can be detected in vivo and monitored over time with the use of transcranial Doppler techniques. If a PFO is present, its closure before surgical manipulation of the fracture is feasible and could have important protective effects against massive systemic embolization. PMID- 11890851 TI - Diffusion-weighted and gradient echo magnetic resonance findings of hemichorea hemiballismus associated with diabetic hyperglycemia: a hyperviscosity syndrome? AB - BACKGROUND: The magnetic resonance (MR) imaging findings of hemichorea hemiballismus (HCHB) associated with hyperglycemia are characterized by hyperintensities in the striatum on T1-weighted MR images and computed tomographic scans, with a mechanism of petechial hemorrhage considered to be responsible. Diffusion-weighted MR imaging (DWI) has been reported to detect early ischemic damage (cytotoxic edema) as bright areas of high signal intensity and vasogenic edema as areas of heterogeneous signal intensity. We report various DWI findings in 2 patients with hyperglycemic HCHB. OBJECTIVES: To describe the DWI and gradient echo findings and characterize the types of edema in HCHB associated with hyperglycemia. SETTING: A tertiary referral center neurology department. DESIGN AND METHODS: Two patients with HCHB associated with hyperglycemia underwent DWI, gradient echo imaging, and conventional MR imaging with gadolinium enhancement. The patients had an elevated serum glucose level on admission and a long history of uncontrolled diabetes, and the symptoms were controlled by dopamine receptor blocking agents. Initial DWIs were obtained 5 to 20 days after symptom onset. Apparent diffusion coefficient (ADC) values were measured in the abnormal lesions with visual inspection of DWI and T2-weighted echo planar images. RESULTS: T1- and T2-weighted MR images and brain computed tomographic scans showed high signal intensities in the right head of the caudate nucleus and the putamen. Gradient echo images were normal. The DWIs showed bright high signal intensity in the corresponding lesions (patient 1), and the ADC values were decreased. The decrease in ADC and the high signal intensity on DWI persisted despite the disappearance of HCHB, even after 70 days. CONCLUSIONS: Gradient echo MR imaging findings were normal in HCHB with hyperglycemia, whereas DWI and the ADC map showed restricted diffusion, which suggests that hyperviscosity, not petechial hemorrhage, with cytotoxic edema can cause the observed MR abnormalities. PMID- 11890853 TI - Atypical brainstem encephalitis caused by herpes simplex virus 2. AB - BACKGROUND: Herpes simplex encephalitis is one of the most common and serious sporadic encephalitides of immunocompetent adults. Herpes simplex virus 2 (HSV-2) infections of the central nervous system usually manifest as subacute encephalitis, recurrent meningitis, myelitis, and forms resembling psychiatric syndromes. OBJECTIVES: To report and discuss magnetic resonance imaging (MRI) findings and clinical features in atypical brainstem encephalitis and facial palsy associated with HSV-2. SETTING: Neurology department of a tertiary referral center. PATIENT: A 37-year-old woman was admitted to the hospital with fever, diplopia, left hemiparesis, sensory change in the face and limbs, personality changes, frontal dysexecutive syndrome, and a stiff neck. Brain MRI showed multifocal high-signal intensities in the pons, midbrain, and frontal lobe white matter on T2-weighted and fluid-attenuated inversion recovery images. Cerebrospinal fluid (CSF) polymerase chain reaction (PCR) amplification analysis was positive for HSV-2. Acyclovir therapy was started, and the encephalitic symptoms disappeared with a negative conversion of HSV-2 PCR in the CSF. However, after the discontinuation of acyclovir therapy, peripheral facial palsy occurred on the left side. A possible relapse or delayed manifestation of the HSV-2 infection was suspected, and the acyclovir therapy was restarted. A complete remission was achieved 3 days after the treatment. She was discharged without any neurologic sequelae. CONCLUSIONS: We describe a patient who developed atypical encephalitis due to HSV-2 and peripheral facial palsy, which could also be related to the HSV-2. This case suggests that HSV-2 should be considered among the possible causes of atypical or brainstem encephalitis and that the PCR amplification method of the CSF can help reveal the possible cause of HSV-2. PMID- 11890854 TI - Familial diffuse Lewy body disease, eye movement abnormalities, and distribution of pathology. AB - BACKGROUND: Familial diffuse Lewy body disease (DLBD) is rare and not yet associated with a defect in the synuclein gene. In the differential diagnosis of the parkinsonian syndromes, defects in vertical gaze tend to be identified with progressive supranuclear palsy. False-positive diagnosis of progressive supranuclear palsy can occur, and defects in vertical gaze have been reported in DLBD, although so far a pure vertical gaze palsy associated with pathological abnormalities in the substrate for vertical gaze has not been described. OBJECTIVES: To report the clinical and pathological findings in 2 siblings with DLBD, and to relate the distribution of the pathological abnormalities in the brainstem to centers for vertical gaze. MATERIALS: For several years, 2 Irish siblings experienced a progressive parkinsonism-dementia complex associated in one with a defect in vertical gaze and in both with visual hallucinations. RESULTS: In both patients, results of pathological examination revealed (1) Lewy bodies positive for ubiquitin and alpha-synuclein together with cell loss and gliosis in the substantia nigra, locus ceruleus, and neocortex; and (2) similar findings in the rostral interstitial nucleus of the medial longitudinal fasciculus, the posterior commissure, and the interstitial nucleus of Cajal (substrates for vertical gaze). CONCLUSIONS: Familial DLBD (not shown to be genetically as distinct from environmentally transmitted) has been shown to exist in an Irish family. Caution should be enjoined in the interpretation of defects in vertical gaze in the differential diagnosis of the parkinsonian syndromes. PMID- 11890856 TI - Unequal crossing-over in unique PABP2 mutations in Japanese patients: a possible cause of oculopharyngeal muscular dystrophy. AB - BACKGROUND: Oculopharyngeal muscular dystrophy (OPMD) is an adult-onset autosomal dominant muscle disease with a worldwide distribution. Recent findings reveal the genetic basis of this disease to be mutations in the polyA binding-protein 2 (PABP2) gene that involve short expansions of the GCG trinucleotide repeat encoding a polyalanine tract. The underlying mechanism causing the triplet expansion mutation in PABP2 remains to be elucidated, although the DNA slippage model is thought to be a plausible explanation of that. METHODS AND RESULTS: We analyzed PABP2 using polymerase chain reaction analysis and DNA sequencing in Japanese patients with pathologically confirmed OPMD, and found mutated (GCG)(6)GCA(GCG)(3)(GCA)(3)GCG and (GCG)(6)(GCA)(3)(GCG)(2)(GCA)(3)GCG alleles instead of the normal (GCG)(6)(GCA)(3)GCG allele. These mutated alleles could be explained by the insertions or duplications of (GCG)(3)GCA and (GCG)(2)(GCA)(3), respectively, but not by the simple expansion of GCG repeats. The clinical features of our patients were compatible with those of other Japanese patients carrying PABP2 that encodes a polyalanine tract of the same length, but were not compatible with those of Italian patients. CONCLUSIONS: The mutated alleles identified in our Japanese patients with OPMD were most likely due to duplications of (GCG)(3)GCA and (GCG)(2)(GCA)(3) but not simple expansions of the GCG repeats. Therefore, unequal crossing-over of 2 PABP2 alleles, rather than DNA slippage, is probably the causative mechanism of OPMD mutations. All mutations that have been reported in patients with OPMD so far can be explained with the mechanism of unequal crossing-over. On the other hand, comparison of the clinical features of our patients with those of other patients in previous reports suggests that specific clinical features cannot be attributed to the length of the polyalanine tract per se. PMID- 11890855 TI - Recurrent stroke as a manifestation of primary angiitis of the central nervous system in a patient infected with human immunodeficiency virus. AB - CONTEXT: Cerebral vasculitis in patients infected with human immunodeficiency virus (HIV) is usually related to additional or secondary infectious agents other than neoplastic diseases or HIV itself. OBJECTIVE: To describe a 31-year-old patient infected with HIV who presented with 2 recurrent, acute episodes of neurologic impairment in a 5-month period. DESIGN: Comparison of clinical and histologic data between the present case and previously published cases. SETTING: Community hospital. PATIENT: A 31-year-old, HIV-infected patient with recurrent strokes and chronic lymphocytic meningitis. INTERVENTION: After ruling out cardiac embolisms and coagulation disorders, the presence of central nervous system vasculitis, probably secondary to an infectious process, was suspected based on the clinical examination and cerebrospinal fluid abnormalities. RESULTS: Necropsy findings suggest the diagnosis of primary angiitis of the central nervous system, and the only infectious agent that could be found was HIV. CONCLUSIONS: Histologic studies were compatible with a diagnosis of primary angiitis of the central nervous system, but the pathogenic role of HIV in the genesis of the vasculitic process cannot be elucidated. PMID- 11890857 TI - A basilar tip aneurysm unsuitable for therapy: noninvasive visualization with multislice computed tomographic angiography. PMID- 11890858 TI - An ancient eye. PMID- 11890859 TI - Temporal lobectomy. AB - Temporal lobectomy is the most common neurosurgical procedure performed for medically refractory epilepsy. Temporal lobe epilepsy has historically been one of the most difficult forms of epilepsy to treat, and it carries a significant psychosocial and medical burden. The inadequacy of medical therapy led to the development of effective neurosurgical procedures, and this article cites several seminal authors who pioneered the development of the field. PMID- 11890862 TI - Factors associated with dementia and cognitive impairments in veterans with human immunodeficiency virus. PMID- 11890860 TI - Cognitive decline after coronary artery surgery. PMID- 11890863 TI - Dendritic cells for treatment of human malignant brain tumours. PMID- 11890864 TI - Current status of adoptive immunotherapy of malignancies. AB - Adoptive immunotherapy involves the transfer of immune effectors with antitumour activity into the tumour bearing host. Early approaches such as lymphokine activator killer (LAK) cells and tumour infiltrating lymphocytes (TILs) have yielded occasional clinical responses. More recently, attempts to stimulate and/or select antigen-specific T-cells in vitro have demonstrated that tumour specific adoptive immunotherapy is possible. These approaches require complicated and time consuming in vitro stimulation procedures. Therefore, genetic modification of bulk T-cell populations is an attempt to create a large population of T-cells with a single specificity. In addition to work being done to develop the most potent effector, other studies are working on improving T cell trafficking to tumours and interfering with the tumour-induced immunosuppression that can impair in vivo T-cell activity. PMID- 11890865 TI - Cell-based therapy approaches using dying cells: from tumour immunotherapy to transplantation tolerance induction. AB - Cell-based therapies are promising approaches to treat uncontrolled pathologies, such as tumours. Apoptotic tumour cells have recently been proposed as a source of tumour-associated antigens to stimulate an efficient immune response. However, a complex relationship exists between apoptosis and the immune system. In this review, the different factors that may influence immune responses against apoptotic cells are detailed and discussed in the light of recent publications. These factors include the nature of the phagocytes and the receptors involved in apoptotic cell uptake, as well as the environment in which cells are dying. A possible distinction between apoptosis and necrosis by immune system sentinels adds a further level of complexity. The potential use of the immunomodulatory properties associated with apoptosis to favour engraftment and induce tolerance in transplantation is then discussed. In conclusion, this review will suggest appropriate conditions to efficiently and safely use apoptotic cells as a new cell therapy product. PMID- 11890866 TI - The development of immunotherapies for non-small cell lung cancer. AB - Standard of care for non-small cell lung cancer (NSCLC) (surgery, chemotherapy and radiation) may enhance patient survival but the enhancement is typically transient and quite uncommon with advanced disease. Researchers and medical professionals are using new approaches to improve patient mortality and morbidity. One of these approaches, immunotherapy, seeks to stimulate antitumour immunity above a threshold level needed for tumour regression or to induce stability in the face of progression. Among the most established approaches are vaccines involving monoclonal antibodies (mAbs) or immune effector cells. These approaches stimulate the humoral and cell-mediated arms of the immune system, respectively. As the development of humanised or fully human antibodies has spurred exploration of radioimmunoconjugates and immunotoxins, mAbs have enjoyed a revival of sorts. Cell-based therapies using the tumour cell itself as a vaccine component has resulted in disease stabilisation or regression. In addition, immune cells (e.g., T-lymphocytes and dendritic cells [DCs]) are the focal point of numerous patient trials in which meaningful clinical impact was achieved. In general, there are many tactics under development for the treatment of NSCLC. This review primarily concerns immunotherapeutic cancer treatments that are either already in clinical trial or well progressed into preclinical studies. PMID- 11890867 TI - Immunomodulators in the treatment of cutaneous lymphomas. AB - Cutaneous lymphomas (CLs) are a heterogeneous group of lymphoproliferative cutaneous neoplasms that in many cases cannot be cured by standard radio- and chemotherapy. Immunotherapy holds promise for a well-tolerated treatment approach that allows long-time disease control. Unspecific immunotherapy such as IFNs is successfully used in cutaneous T-cell lymphomas (CTCLs). Antibiotic treatment is often effective in cutaneous B-cell lymphomas (CBCLs). New treatment approaches include fusion toxins and chimeric monoclonal antibodies (mAbs) such as rituximab for CBCLs. The identification of tumour antigens in CTCLs renders these diseases susceptible for vaccination approaches. PMID- 11890868 TI - Developments in the area of bioadhesive drug delivery systems. AB - This paper aims to review the current progress in bioadhesion for drug delivery applications as well as new techniques related to this field. Research started with mucoadhesive polymers that had already been in use as excipients and were rapidly used in new formulations. Their major drawback was found in their unspecific binding, as they adhere to almost any mucosal surface. As some of the polymers showed additional properties such as enzyme inhibition and permeation enhancement, however, they remain interesting as multifunctional excipients. In contrast to mucoadhesion, the concept of specific bioadhesion by use of lectins and other adhesion molecules is now gaining increasing attention as these substances bind directly to receptors on the cell surface rather than to the mucus gel layer. Since specific binding to the cell surface is often followed by uptake and intracellular transport, new chances for drug delivery evolved. Bioadhesion may, thus, enable researchers to deliver macromolecular drugs directly to specific target cells and has implications also relevant to other fields of science, such as tissue engineering, gene delivery and nanotechnology. PMID- 11890869 TI - Genetic therapies and xenotransplantation. AB - The number of patients in need of an organ transplant is increasing, while the number of satisfactory sources of organs has declined in many countries [101]. The resulting shortage of human organs has spurred an urgent effort to investigate alternative therapies, including the use of animal organs, tissues and cells (i.e., xenotransplantation). Advances in genetic engineering have provided essential tools for the development of practical solutions to human disease. The area of xenotransplantation is no exception. In fact, the use of genetic therapies is especially attractive in the transplant setting as it offers an opportunity to manipulate the donor tissue rather than the recipient. This review will describe the obstacles in the clinical application of xenotransplantation and how genetic engineering might be used to address them. PMID- 11890870 TI - Head and neck cancer: gene therapy approaches. Part II: genes delivered. AB - In Part I, the review summarised the safety of adenoviral vectors and provided insight into approaches being undertaken to improve the specificity, durability and potency of adenoviral delivery vehicles. In Part II, brief discussions are held regarding results of preclinical and clinical trials with a variety of different genes, which have demonstrated antitumour activity in squamous cell carcinoma of the head and neck region (HNSCC). Studies have been performed with a variety of immune modulatory genes. Preliminary results demonstrate activity with several cytokine genes, tumour antigen genes and co-stimulatory molecule genes. Despite only preliminary results, thus far, a theoretical attractive feature for the use of gene therapy for the enhancement of immune modulation is that local injection of the gene product appears to be well tolerated. It is also successful in inducing systemic immune response, potentially providing effect to metastatic sites distal from the injected site. Animal studies have confirmed efficacy in the use of specific targeting of molecules regulating cancer growth (EGF receptor [EGFR], super oxide dismutase [SOD], cyclin D1, E1A and Bcl-2). These approaches are discussed. However, the most significant clinical advances for the use of gene therapy in advanced HNSCC involves two agents: Adp53 and ONYX-015. Preliminary Phase I and II results suggest evidence of efficacy and justify accrual Phase III trials, which are currently ongoing. PMID- 11890871 TI - Advances in the management of Anderson-Fabry disease: enzyme replacement therapy. AB - Anderson-Fabry disease (AFD) is a lysosomal storage disorder (LSD) due to alpha galactosidase A (alpha-Gal A) deficiency and the resultant accumulation of incompletely metabolised glycosphingolipids (GSLs), primarily globotriosylceramide (Gb(3)), within various tissues. It is an X-linked multisystem disorder characterised by progressive renal insufficiency, with added morbidity from cardio- and cerebrovascular involvement, and associated with significant impact on quality of life and diminished lifespan. The disease manifests primarily in hemizygous males; however, there is increasing recognition that heterozygous (carrier) females may also develop disease-related complications, although onset among affected women may be delayed. Until recently, treatment has been limited to symptomatic management of pain and other measures to alleviate the problems associated with end-stage complications from renal, cardiac and nervous system involvement. The availability of the recombinant enzyme offers the potential of a safe and effective targeted treatment approach. At the moment, two distinct enzyme formulations are approved in Europe (and in other countries) and both continue to undergo FDA evaluation in the US. Increasing knowledge of the natural history of AFD and greater experience with enzyme therapy should enable optimal patient care. The relative rarity and complexity of AFD necessitates a multi-disciplinary team approach that may be facilitated by a centralised registry. PMID- 11890872 TI - M-Vax: an autologous, hapten-modified vaccine for human cancer. AB - A novel approach to active immunotherapy has been devised based on modification of autologous cancer cells with the hapten, dinitrophenyl (DNP). This technology is being developed by AVAX Technologies as a treatment for melanoma under the brand name, M-Vax(TM). The treatment program consists of multiple intradermal injections of DNP-modified autologous tumour cells mixed with Bacille Calmette Guerin (BCG). DNP-vaccine administration to patients with metastatic melanoma induces a unique reaction - the development of inflammation in metastatic masses. The inflammation is mediated by IFN-gamma-producing T-lymphocytes, some of which represent expansion of novel clones. Following DNP-vaccine treatment, almost all patients develop delayed-type hypersensitivity (DTH) to autologous, DNP-modified melanoma cells; approximately half also exhibit DTH to autologous, unmodified tumour cells. The toxicity of the vaccine is mild, consisting mainly of papules or pustules at the injection sites. Clinical trials have been conducted in two populations of melanoma patients: stage IV with measurable metastases and clinical stage III patients, rendered tumour-free by lymphadenectomy. In 83 patients with measurable metastases, there were 11 antitumour responses: two complete responses (CRs), four partial responses (PRs) and five mixed. Both CRs and two of four PRs occurred in patients with lung metastases. In 214 stage III patients the 5-year overall survival rate was 46% (one nodal site = 48%, in transit metastases = 50%, two nodal sites = 36%). In both populations, the induction of DTH to unmodified autologous tumour cells was associated with significantly longer survival. This technology is applicable to other human cancers and clinical trials have been initiated with ovarian adenocarcinoma. There appear to be no insurmountable impediments to applying this approach to much larger numbers of patients or to developing it as a standard cancer treatment. PMID- 11890873 TI - Methotrexate shows marginal clinical efficiency in early scleroderma. PMID- 11890875 TI - Evaluation and management of pulmonary fibrosis in scleroderma. AB - Pulmonary fibrosis causes significant morbidity and mortality in patients with scleroderma. Lung inflammation identifies patients at greater risk for decline in forced vital capacity and diffusing capacity for carbon monoxide. Factors that are increased in patients with scleroderma with lung fibrosis include connective tissue growth factor, KL-6, pulmonary surfactant-D, tissue inhibitor of metalloproteinase 2, monocyte chemotactic protein-1, macrophage inhibitory protein-1 alpha, soluble interleukin-6 receptors, anti-endothelial cell antibodies, and anti-DNA topoisomerase I antibodies. Potential mechanisms of lung damage in scleroderma include increased production of profibrotic type 2 cytokines and abnormal signaling by thrombin of tenascin-C production by lung fibroblasts, with protein kinase C epsilon as an intermediate in the signaling pathway. Treatment of scleroderma lung disease with cyclophosphamide may have a beneficial effect on pulmonary function and survival. Lung transplantation provides a therapeutic option for patients with scleroderma with end-stage lung disease. PMID- 11890874 TI - The genetics of systemic sclerosis. AB - The etiopathogenesis of systemic sclerosis (SSc) is unclear. With no definitive evidence supporting an environmental cause, recent attention has focused on genetic factors. Familial clustering and ethnic influences have been demonstrated. Human leukocyte antigen (HLA) associations exist but are more related to the presence of particular autoantibodies rather than to the disease. In addition, no single major histocompatibility complex (MHC) allele predisposes to SSc in all ethnic groups. The role of microchimerism in SSc is a novel yet unproven hypothesis that may be related to intergenerational HLA compatibility. Recent studies investigating polymorphisms in genes coding for extracellular matrix proteins and cell-signaling molecules implicate non-MHC areas in SSc pathogenesis. The data reviewed suggest that SSc is a multigenic complex disorder. PMID- 11890877 TI - Raynaud's phenomenon: epidemiology and risk factors. AB - Although originally described more than 100 years ago, the pathophysiology of Raynaud's phenomenon (RP) remains incompletely understood. Epidemiologic studies have the potential to improve our understanding of the pathogenesis of RP and to identify potential targets for therapeutic interventions. Such studies require standardized instruments to accurately identify subjects with RP. Dr. Maricq was the first to develop and validate a standardized instrument to classify patients with this disorder. Her work has facilitated many of the epidemiologic studies of RP that are discussed in this review. PMID- 11890876 TI - Scleroderma-like cutaneous syndromes. AB - Several distinct entities associated with dermal fibrosis can mimic scleroderma/systemic sclerosis. The list of scleroderma-like conditions or scleroderma variants includes eosinophilic fasciitis, localized forms of scleroderma, scleredema and scleromyxedema, keloids, and environmental exposure associated conditions including eosinophilia-myalgia syndrome and pseudosclerodermas induced by various drugs. Although these conditions are relatively uncommon, their accurate recognition is essential to avoid misdiagnosis and inappropriate therapy. The pathogenesis of these scleroderma variants appears to share similarities with each other and with that of scleroderma. Better understanding of scleroderma-like disorders is emerging through epidemiologic investigations, and in vivo and in vitro experimental research. Activation of eosinophils and disordered regulation of fibroblast collagen synthesis, apoptosis, and proliferation are recurrent findings in these disorders. The etiologic role of infection with Borrelia species or other microorganisms remains controversial. Cytokines such as transforming growth factor-beta, interleukin-4, interleukin-13, and connective tissue growth factor contribute to fibrosis in these disorders by inducing an accentuated and persistent fibrogenic response to tissue injury. The role of genetic factors in susceptibility and clinical expression of scleroderma-like conditions remains to be systematically addressed. Because of the relative rarity of these conditions, few well-controlled clinical treatment trials have been performed. In addition, there is no consensus on optimal management. Much anecdotal information and small clinical series indicate that phototherapy may have a role in the treatment of scleroderma-like conditions. PMID- 11890878 TI - Apoptosis and myofibroblasts in the pathogenesis of systemic sclerosis. AB - Tissue fibrosis is the result of a complex series of events focusing on regulation of fibroblast proliferation, synthesis of extracellular matrix, and apoptosis. Transforming growth factor-beta is important for the stimulation of the fibrotic response by promoting the production of extracellular matrix proteins, by promoting the differentiation of the myofibroblast cell morphology, and by protecting these cells against apoptotic stimuli. Other cytokines such as interleukin-1 may have stimulatory and counter-regulatory effects on fibrosis. The effects of these signaling molecules depend on cellular environment and are organ specific. Furthermore, intercellular interactions and cell-matrix interactions can stimulate or inhibit the apoptotic pathway. Through selective inhibition of apoptosis in myofibroblasts, fibrosis can become dysregulated and lead to diseases such as systemic sclerosis. PMID- 11890879 TI - Connective tissue growth factor: a new and important player in the pathogenesis of fibrosis. AB - Connective tissue fibrosis is the final common pathogenic process for almost all forms of chronic tissue injury. Whether caused by vascular dysfunction, inflammation, metabolic injury, trauma, or environmental agents, once initiated the fibrogenic process results in the progressive replacement of the normal tissue architecture with fibrotic lesions that eventually lead to organ compromise and failure. Fibrosis can be considered as a dysregulation in the normal tissue repair mechanism, resulting in severe tissue scarring. Fibrosis appears to be a consequence of linked processes, including the proliferation of resident fibroblast cell types, the increased production and deposition of extracellular matrix components, and the transition of fibroblasts into cells exhibiting a myofibroblast phenotype. Although transforming growth factor-beta (TGF beta) has long been regarded as a pivotal growth factor in the formation and maintenance of connective tissues and as a major driving influence in many progressive fibrotic diseases, attention has focused recently on the role of connective tissue growth factor (CTGF) in fibrosis. CTGF is selectively and rapidly induced in mesenchymally derived cells by the action of TGF beta. CTGF expression is increased in many fibrosing diseases. In addition, increasing evidence from in vivo and in vitro models of tissue remodeling and fibrosis suggest that CTGF may represent a downstream effector molecule of the profibrotic activities of TGF beta in the maintenance and repair of connective tissues and within fibrotic disease settings. PMID- 11890880 TI - Control of connective tissue gene expression by TGF beta: role of Smad proteins in fibrosis. AB - Transforming growth factor-beta (TGF beta) plays a critical role in the development of tissue fibrosis. Its expression is consistently elevated in affected organs and correlates with increased extracellular matrix deposition. During the last few years, tremendous progress has been made in understanding the molecular aspects of intracellular signaling downstream of the TGF beta receptors. In particular, Smad proteins--TGF beta receptor kinase substrates that translocate into the cell nucleus to act as transcription factors--have been studied extensively. Their role in the modulation of extracellular matrix gene expression is discussed in this review. PMID- 11890882 TI - Reflex sympathetic dystrophy. AB - Reflex sympathetic dystrophy (RSD) is composed of five major features: pain, swelling, autonomic dysregulation, movement disorders, and atrophy and dystrophy. RSD is caused by an injury to a specific nerve or the C- and A-delta fibers that innervate the involved tissue. It is a progressive illness that spreads with time and may encompass the entire body. There is no psychological disposition to the problem, but all patients are severely depressed because of the constant pain, lack of sleep, and complete disruption of their lifestyle. The continuing pain is usually secondary to the process of central sensitization. The autonomic dysregulation has a major central nervous system component. Atrophy and dystrophy are partly due to loss of nutritive blood supply to the affected tissues. The movement disorder is partly due to deficiency of GABAergic mechanisms; the tremor is an exaggeration of the normal physiologic tremor. Treatment consists of decreasing the afferent pain, maintaining barrage from the underlying defect, and blocking the sympathetic component of the process. New developments include the use of neurotrophic factors to reverse the phenotypic changes that occur in the dorsal horn and the use of pharmacologic agents to block the activity-dependent NMDA channels that appear to be instrumental in maintaining central sensitization. PMID- 11890881 TI - Animal models for scleroderma: an update. AB - Scleroderma is a progressive debilitating fibrosing disease that may involve multiple organs. The pathogenesis of this disease remains unclear. Animal models for scleroderma are valuable for studying the pathogenesis of this complex disorder and for testing potential treatments for human scleroderma. There are several animal models available that exhibit important features of scleroderma, each with an emphasis on different aspects of the disease (tissue fibrosis, inflammation, vascular injury, or immunologic changes). These models can be separated into several categories in which fibrosis is induced by external agents (vinyl chloride, bleomycin), by breeding of mutant strain combinations (integrin alpha 1 null mouse, MRL/lpr gamma R-/- mouse), and by transplantation of disparate immune cells (sclerodermatous graft versus host disease). In addition, there are spontaneous mutations (UCD 200 chicken, tight skin mouse) in which fibrosis occurs. The tight skin mouse has been reviewed recently. This review discusses the other animal models and some interventions in each. PMID- 11890883 TI - Nonpulmonary manifestations of sarcoidosis. AB - Sarcoidosis is one in a heterogeneous family of granulomatous disorders. The clinical manifestations of sarcoidosis can vary widely, depending on the patient and the tissues involved. Recent advances in our understanding of the immunologic steps leading to granuloma formation and persistence have yet to translate into more effective care for patients with this disease. This review discusses the immunology of granuloma formation and systemic disease, the various nonpulmonary expressions of the clinical disease, and the treatment options available to the practicing physician. PMID- 11890884 TI - Rheumatic manifestations of primary hyperparathyroidism and parathyroid hormone therapy. AB - A frequent manifestation of severe primary hyperparathyroidism was the bone disease osteitis fibrosa cystica. Rarely, excess parathyroid hormone (PTH) was associated with gout and calcium pyrophosphate crystal deposition disease. Surgical cure of primary hyperparathyroidism was occasionally associated with pseudogout. Today, primary hyperparathyroidism is generally asymptomatic. Clinically overt rheumatologic and skeletal effects are mainly of historical interest. Skeletal disease is still detectable by more sensitive techniques. In certain circumstances, PTH may be protective and anabolic for the skeleton. PMID- 11890885 TI - Human milk oligosaccharides: an enzymatic protection step simplifies the synthesis of 3'- and 6'-O-sialyllactose and their analogues. AB - We describe a chemo-enzymatic synthesis of 3'- and 6'-O-sialyllactose, two trisaccharides occurring in the 'acidic fraction' of the human milk oligosaccharides and endowed with potential antiadhesive activity. The key step is the highly regioselective 6'-O-acylation of benzyllactoside, which gave access to suitably protected lactose building blocks to be used as acceptors in the sialylation reaction. Moreover, the synthesis of the carboxymethyl and sulfo analogues of the title compounds is reported. PMID- 11890886 TI - Synthesis of oligosaccharide derivatives related to those from sanqi, a Chinese herbal medicine from Panax notoginseng. AB - Oligosaccharide derivatives from sanqi, a Chinese herbal medicine derived from Panax notoginseng, methyl beta-D-galactopyranosyl-(1-->3)-[alpha-L arabinofuranosyl-(1-->6)]-alpha-D-galactopyranoside, diosgenyl beta-D galactopyranosyl-(1-->3)-[alpha-L-arabinofuranosyl-(1-->6)]-alpha-D galactopyranoside, and methyl beta-D-galactopyranosyl-(1-->3)-[alpha-L arabinofuranosyl-(1-->6)]-alpha-D-galactopyranosyl-(1-->4)-beta-D galactopyranosyl-(1-->3)-[alpha-L-arabinofuranosyl-(1-->6)]-alpha-D galactopyranoside, were synthesized under standard glycosylation conditions. An unexpected alpha-(1-->4) linkage was formed predominantly in the presence of neighboring participation group during regioselective synthesis of hexasaccharide via 3+3 strategy. PMID- 11890887 TI - Synthesis and characterization of water-soluble hydroxybutenyl cyclomaltooligosaccharides (cyclodextrins). AB - We have examined the synthesis of hydroxybutenyl cyclomaltooligosaccharides (cyclodextrins) and the ability of these cyclodextrin ethers to form guest-host complexes with guest molecules. The hydroxybutenyl cyclodextrin ethers were prepared by a base-catalyzed reaction of 3,4-epoxy-1-butene with the parent cyclodextrins in an aqueous medium. Reaction byproducts were removed by nanofiltration before the hydroxybutenyl cyclodextrins were isolated by co evaporation of water-EtOH. Hydroxybutenyl cyclodextrins containing no unsubstituted parent cyclodextrin typically have a degree of substitution of 2-4 and a molar substitution of 4-7. These hydroxybutenyl cyclodextrins are randomly substituted, amorphous solids. The hydroxybutenyl cyclodextrin ethers were found to be highly water soluble. Complexes of HBen-beta-CD with glibenclamide and ibuprofen were prepared and isolated. In both cases, the guest content of the complexes was large, and a significant increase in the solubility of the free drug was observed. Dissolution of the complexes in pH 1.4 water was very rapid, and significant increases in the solubility of the free drugs were observed. Significantly, after reaching equilibrium concentration, a decrease in the drug concentration over time was not observed. PMID- 11890888 TI - Addition of maltodextrins to the nonreducing-end of acarbose by reaction of acarbose with cyclomaltohexaose and cyclomaltodextrin glucanyltransferase. AB - New kinds of acarbose analogues were synthesized by the reaction of acarbose with cyclomaltohexaose and cyclomaltodextrin glucanyltransferase (CGTase). Three major CGTase coupling products were separated and purified by Bio-Gel P2 gel-permeation chromatography. Digestion of the three products by beta-amylase and glucoamylase showed that they were composed of maltohexaose (G6), maltododecaose (G12), and maltooctadecaose (G18), respectively, attached to the nonreducing-end of acarbose. 13C NMR of the glucoamylase product (D-glucopyranosyl-acarbose) showed that the D-glucose moiety was attached alpha- to the C-4-OH group of the nonreducing-end cyclohexene ring of acarbose, indicating that the maltodextrins were attached alpha-(1-->4) to the nonreducing-end cyclohexene of acarbose. PMID- 11890889 TI - Transglucosidation of methyl and ethyl D-glucopyranosides by alcoholysis. AB - The transglucosidations of methyl 4-O-methyl-alpha- and -beta-D-glucopyranoside in ethanolic camphor-10-sulfonic acid, and of ethyl 4-O-methyl-alpha- and -beta-D glucopyranoside in methanolic camphor-10-sulfonic acid, have been studied. Samples were removed at intervals and the proportions of the glucosides determined by GC of their acetates. The results show that the anomer with the inverted configuration predominates in the initially formed product (approximately 59-70%). This indicates that all the studied reactions proceed via the same mechanism, involving exocyclic C-O cleavage and formation of a glucopyranosylium ion, but that the eliminated alcohol exerts some steric hindrance, which favors the approach of the other alcohol from the opposite side. PMID- 11890890 TI - Effect of calcium ions on the organization of iota-carrageenan helices: an X-ray investigation. AB - X-ray fiber diffraction analysis confirms that calcium iota-carrageenan forms a threefold, right-handed, half-staggered, parallel, double helix of pitch 26.42 A stabilized by interchain hydrogen bonds. According to the detailed structural results, three helices are packed in a trigonal unit cell (a=23.61 and c=13.21 A). Strong interactions between the sulfate groups of neighboring helices, mediated by calcium ions and water molecules, are responsible for stabilizing the three-dimensional structure. PMID- 11890891 TI - Crystal structure of the cyclomaltohexaose (alpha-cyclodextrin) complex with isosorbide dinitrate. Guest-modulated channel-type structure. AB - The crystal structure of the 2:1 complex of cyclomaltohexaose (alpha cyclodextrin, alpha-CD) with isosorbide dinitrate was determined by single crystal X-ray analysis. In the crystal with the space group C2, two cyclomaltohexaose molecules form a head-to-head dimer with the secondary hydroxy group sides facing each other. The dimer unit is stacked along the crystallographic c-axis to form a channel-type structure. The isosorbide dinitrate molecule is encapsulated in the cylindrical cavity of the cyclomaltohexaose dimer. The dimeric structure exhibits pseudo twofold symmetry, and the guest molecule is disordered on the local symmetry axis. The isosorbide moiety is located at the center of the dimer cavity, and the nitrate groups penetrate into the cyclomaltohexaose rings. The guest molecule modulates the dimer structure to attain the most stable accommodation into the cavity. The cyclomaltohexaose molecules are laterally shifted away from each other to create the cavity fitted to the shape of the guest molecule. As the result, the intermolecular hydrogen bonds between secondary hydroxy-groups are not fully formed, but the dimeric structure is stabilized by the interaction with the guest molecule. PMID- 11890893 TI - Quantitative production of 2-acetamido-2-deoxy-D-glucose from crystalline chitin by bacterial chitinase. AB - Finely powdered alpha- and beta-chitin can be completely hydrolyzed with chitinase (EC 3.2.1.14) and beta-N-acetylhexosaminidase (EC 3.2.1.52) for the production of 2-acetamido-2-deoxy-D-glucose (GlcNAc). Crude chitinase from Burkholderia cepacia TU09 and Bacillus licheniformis SK-1 were used to digest alpha- and beta-chitin powder. Chitinase from B. cepacia TU09 produced GlcNAc in greater than 85% yield from beta- and alpha-chitin within 1 and 7 days, respectively. B. licheniformis SK-1 chitinase completely hydrolyzed beta-chitin within 6 days, giving a final GlcNAc yield of 75%, along with 20% of chitobiose. However, only a 41% yield of GlcNAc was achieved from digesting alpha-chitin with B. licheniformis SK-1 chitinase. PMID- 11890892 TI - Monte Carlo docking simulations of cyclomaltoheptaose and dimethyl cyclomaltoheptaose with paclitaxel. AB - The molecular basis for the remarkable enhancement of the solubility of paclitaxel by O-dimethylcyclomaltoheptaose (DM-beta-CD) over cyclomaltoheptaose (beta-cyclodextrin, beta-CD) was investigated with Monte Carlo docking minimization simulation. As possible guests of inclusion complexation for the host cyclic oligosaccharides, two functional moieties of the suggested solution structure of paclitaxel were used where one is the C-3'N benzoyl moiety (B-ring) and the other is a hydrophobic (HP) cluster site among the C-3' phenyl (C-ring), C-2 benzoate (A-ring), and C-4 acetoxy moieties. The energetic preference of inclusion complexation of DM-beta-CD over beta-CD was analyzed on the basis of more efficient partitioning process of DM-beta-CD into the hydrophobic cluster site of the paclitaxel. PMID- 11890894 TI - Depolymerization of N-succinyl-chitosan by hydrochloric acid. AB - N-Succinyl-chitosan (1) was depolymerized with 7.5 M aqueous HCl at room temperature or 3.3 M aqueous HCl at 40 degrees C and the molecular weights (MW) of the products were determined by size-exclusion chromatography-multi angle light scattering (SEC-MALS) and their viscometric features were investigated. The intrinsic viscosity ([eta]) obtained at the concentration of 0.1-0.3% (w/v) in saline showed a linear relationship between log[eta] and log MW, which provided the coefficients in the Mark-Houwink equation. PMID- 11890895 TI - Confirmation of the structure of tetra-O-(tert-butyldimethylsilyl)-D-glucono-1,4 lactone formed by silylation of D-glucono-1,5-lactone. AB - The structure of tetra-O-(tert-butyldimethylsilyl)-D-glucono-1,4-lactone made by the silylation of D-glucono-1,5-lactone has been confirmed by single-crystal X ray analysis. PMID- 11890897 TI - Regulation of glycogen metabolism in hepatocytes through adenosine receptors. Role of Ca2+ and cAMP. AB - The objective of this work is to identify the adenosine receptor subtype and the triggered events involved in the regulation of hepatic glycogen metabolism. Glycogenolysis, gluconeogenesis, cAMP, and cytosolic Ca2+ ([Ca2+](cyt)) were measured in isolated hepatocytes challenged with adenosine A1, A2A, and A3 receptor-selective agonists. Stimulation of adenosine receptor subtypes with selective agonists in Ca2+ media produced a dose-dependent increase in [Ca2+]cyt with A1>A2=A3, cAMP with A2A, glycogenolysis with A1>A2A>A3, and gluconeogenesis with A2A>A1>A3, in addition, a decrease in cAMP was observed with A1=A3. Comparatively, in Ca2+-free media or with a cell membrane-permeant Ca2+ chelator, activation of these adenosine receptors with the same selective agonists produced a smaller and transient rise in [Ca2+]cyt with A1=A3>A2, no rise in glycogenolysis and gluconeogenesis with A3>A1, but a full rise with A2A. Thus, in isolated rat hepatocytes activation of the adenosine A1 receptor triggered Ca2+ mediated glycogenolysis, activation of the adenosine A2A receptor stimulated cAMP mediated gluconeogenesis, and activation of the adenosine A3 receptor increased [Ca2+]cyt and decreased cAMP with minor changes in glycogen metabolism. PMID- 11890899 TI - FK506 induces chondrogenic differentiation of clonal mouse embryonic carcinoma cells, ATDC5. AB - FK506 (Tacrolimus) and cyclosporin A exert their immunosuppressive effects via a common mechanism, calcineurin inhibition, after binding to intracellular proteins termed immunophilins: FK506-binding protein (FKBP) and cyclophilin. In this study, FK506 was found to induce chondrogenic differentiation of ATDC5 cells (clonal mouse embryonal carcinoma cells) in a concentration-dependent manner (0.1 1000 ng/ml). Immunohistochemical staining showed that ATDC5 cells induced to differentiate by FK506 produced proteoglycan and type II collagen, main components of the extracellular matrix of cartilage. Rapamycin, an immunosuppressant that binds to FKBP, antagonized the effect of FK506. Cyclosporin A did not induce chondrogenesis at concentrations up to 1000 ng/ml. Taken together, these results suggest that FK506 induces chondrogenic differentiation of ATDC5 cells via a calcineurin-independent mechanism, after binding to FKBP. PMID- 11890898 TI - Cyclic AMP inhibits stretch-induced overexpression of fibronectin in glomerular mesangial cells. AB - Glomerular hypertension is proposed to play an important role in the progression of various glomerular diseases. Glomerular mesangial cells are considered to be exposed to the stretch stress due to glomerular hypertension and are found to produce the excess amount of extracellular matrix (ECM) proteins including fibronectin when exposed to the mechanical stretch. Herein, we provide the evidence that cAMP-generating agents inhibit the stretch-induced overexpression of fibronectin through the inhibition of the stretch-induced activation of mitogen-activated protein kinases (MAPKs) in protein kinase-A-dependent manner. We also found that the mechanical stretch enhanced the binding of nuclear extracts to activator protein-1 (AP-1)-like sequences in the promoter region of rat fibronectin gene and this enhancement was also prevented by the cAMP generating agent. These results indicate that the agents, which activate cAMP/protein kinase-A axis, may work protectively against the injury from glomerular hypertension in mesangial cells. PMID- 11890900 TI - Activation of KCNQ5 channels stably expressed in HEK293 cells by BMS-204352. AB - The novel anti-ischemic compound, BMS-204352 ((3S)-(+)-(5-chloro-2-methoxyphenyl) 1,3-dihydro-3-fluoro-6-(trifluoromethyl)-2H-indol-2-one)), strongly activates the voltage-gated K+ channel KCNQ5 in a concentration-dependent manner with an EC50 of 2.4 microM. At 10 microM, BMS-204352 increased the steady state current at -30 mV by 12-fold, in contrast to the 2-fold increase observed for the other KCNQ channels [Schroder et al., 2001]. Retigabine ((D-23129; N-(2-amino-4-(4 fluorobenzylamino)-phenyl) carbamic acid ethyl ester) induced a smaller, yet qualitatively similar effect on KCNQ5. Furthermore, BMS-204352 (10 microM) did not significantly shift the KCNQ5 activation curves (threshold and potential for half-activation, V1/2), as observed for the other KCNQ channels. In the presence of BMS-204352, the activation and deactivation kinetics of the KCNQ5 currents were slowed as the slow activation time constant increased up to 10-fold. The M current blockers, linopirdine (DuP 996; 3,3-bis(4-pyridinylmethyl)-1 phenylindolin-2-one) and XE991 (10,10-bis(4-pyridinylmethyl)-9(10H) anthracenone), inhibited the activation of the KCNQ5 channel induced by the BMS 204352. Thus, BMS-204352 appears to be an efficacious KCNQ channels activator, and the pharmacological properties of the compound on the KCNQ5 channel seems to be different from what has been obtained on the other KCNQ channels. PMID- 11890901 TI - Neuroprotective effects of N-acetylaspartylglutamate in a neonatal rat model of hypoxia-ischemia. AB - Neuroprotective effects of N-acetylaspartylglutamate (NAAG), the precursor of glutamate and a selective agonist at the Group II metabotropic glutamate (mGlu) receptor, against hypoxic-ischemic brain injury were examined in a neonatal rat model of cerebral hypoxia-ischemia. The neonatal hypoxia-ischemia procedure (unilateral carotid artery ligation followed by exposure to an 8% oxygen hypoxic condition for 1.5 h) was performed in 7-day-old rat pups. Following unilateral carotid artery ligation, NAAG (0.5 to 20 mg/kg, i.p.) was administered before or after the hypoxic exposure. Brain injury was examined 1-week later by weight reduction in the ipsilateral brain and by neuron density in the hippocampal CA1 area. In the saline-treated rat, neonatal hypoxia-ischemia resulted in severe brain injury as indicated by a 24% reduction in the ipsilateral brain weight. Low doses of NAAG (2-10 mg/kg, but not 0.5 mg/kg), administered before or even if 1 h after the hypoxic exposure, greatly reduced hypoxia-ischemia-induced brain injury (3.8-14.2% reduction in the ipsilateral brain weight). A high dose of NAAG (20 mg/kg) was ineffective. While L(+)-2-Amino-4-phosphonobutyric acid (L-AP4) and trans-[1S,3R]-1-Amino-cyclopentane-1, 3-dicarboxylic acid (t-ACPD) were unable to provide protection against hypoxic-ischemic brain injury, 2-(phosphonomethyl) pentanedioic acid (2-PMPA), an inhibitor of N-acetylated alpha-linked acidic dipeptidase (NAALADase), which hydrolyzes endogenous NAAG into N-acetyl-aspartate and glutamate, significantly reduced neonatal hypoxia-ischemia-induced brain injury. (alphaS)-alpha-Amino-alpha-[(1S, 2S)-2-carboxycyclopropyl]-9H-xanthine-9 propanoic acid (LY341495), a selective antagonist at the mGlu2/3 receptor, prevented the neuroprotective effect of NAAG. Neuron density data measured in the hippocampal CA1 area confirmed that ipsilateral brain weight reduction was a valid measure for hypoxic-ischemic brain injury. Neonatal hypoxia-ischemia stimulated an elevation of cyclic AMP (cAMP) concentration in the saline-treated rat brain. NAAG, L-AP4 and t-ACPD all significantly decreased hypoxia-ischemia induced elevation of cAMP. LY341495 blocked the effect of NAAG, but not of L-AP4 or t-ACPD, on hypoxia-ischemia-stimulated cAMP elevation. The overall results suggest that the neuroprotective effect of NAAG is largely associated with activation of mGlu2/3 receptor. PMID- 11890903 TI - Blockade of adenosine A1 receptors in the posterior cingulate cortex facilitates memory in rats. AB - Male Wistar rats were bilaterally implanted with indwelling cannulae in the caudal region of the posterior cingulate cortex. After recovery, animals were trained in a step-down inhibitory avoidance task (3.0-s, 0.4-mA foot shock) and received, immediately after training, a 0.5-microl infusion of the adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA; 1, 50 or 100 nM) or of the adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX; 1, 25 or 50 nM). Animals were tested twice, 1.5 h and, again, 24 h after training, in order to examine the effects of these agents on short- and long-term memory, respectively. Only 50-nM DPCPX was effective in altering memory, promoting a facilitation. These results suggest that adenosine A1 receptors in the posterior cingulate cortex inhibit memory consolidation in a way that their blockade facilitates memory for inhibitory avoidance in rats. PMID- 11890902 TI - Erythropoietin prevents cognition impairment induced by transient brain ischemia in gerbils. AB - Erythropoietin has recently been studied for its role in the central nervous system (CNS). It has been shown to exert neuroprotective effects in different models of brain injury. We studied whether neuroprotective effects assessed from the reduction of neuronal loss after transient brain ischemia are associated to the preservation of learning ability. Recombinant human erythropoietin (0.5-25 U) was injected in the lateral cerebral ventricle of gerbils that are subjected to temporary (3 min) bilateral carotid occlusion. Post-ischemic histological evaluation of CA1 area neuronal loss and passive avoidance test were performed. Treatment with recombinant human erythropoietin significantly reduced delayed neuronal death in the CA1 area of the hippocampus and prevented cognition impairment in the passive avoidance test. These data indicate that recombinant human erythropoietin neuroprotective effects in brain ischemia are associated with the preservation of learning function. PMID- 11890904 TI - Interactions of sildenafil with various coronary vasodilators in isolated porcine coronary artery. AB - There are reports of serious hypotension or circulatory shock when sildenafil citrate, a selective cyclic nucleotide phosphodiesterase type 5 inhibitor, which was developed for the treatment of erectile dysfunction, is given to patients taking certain coronary vasodilators. We thus examined the interaction of sildenafil with various coronary vasodilators including nitric oxide (NO) donors in isolated porcine coronary artery. Sildenafil caused concentration-dependent relaxations of the artery precontracted with U46619 (9,11-dideoxy-9 alpha,11 alpha-methanoepoxy-prostaglandin F(2alpha)). Incubation with the NO synthase inhibitor NG-nitro-L-arginine or the soluble guanylate cyclase inhibitor ODQ (1H [1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one) significantly shifted the concentration-response curve for sildenafil to the right without affecting the maximum response, indicating that some part of the relaxant response to sildenafil may be the result of the inhibition of phosphodiestrase type 5-induced degradation of cyclic GMP (cGMP) that is produced through guanylate cyclase activation by NO released spontaneously. The relaxant effects of the vasodilators with an NO donor property, isosorbide dinitrate, sodium nitroprusside, nicorandil and nipradilol, were significantly enhanced by sildenafil, as shown by a significant leftward shift of their concentration-response curves. In contrast, the relaxant responses to the drugs without a property as an NO donor, diltiazem, celiprolol and pinacidil, were not affected by sildenafil. The cGMP level of the tissue was elevated after adding sildenafil, and the cGMP-generating effect of a combination of sildenafil and sodium nitroprusside was higher than that of each drug alone. The cyclic AMP level determined simultaneously was not changed by sildenafil. These results suggest that sildenafil potentiates specifically the relaxant responses of porcine coronary artery to the drugs which behave as an NO donor, providing basic evidence that the benefit of sildenafil in the treatment of erectile dysfunction can be limited by a risk of marked vasodilation when used together with NO-related coronary vasodilators. PMID- 11890905 TI - Selective mitochondrial K(ATP) channel activation results in antiarrhythmic effect during experimental myocardial ischemia/reperfusion in anesthetized rabbits. AB - We investigated the effects of administration of non-hypotensive doses of ATP sensitive K+ channel (K(ATP)) openers (nicorandil and aprikalim), and a specific mitochondrial K(ATP) channel blocker (5-hydroxydecanoate) prior to and during coronary occlusion as well as prior to and during post-ischemic reperfusion on survival rate, ischemia/reperfusion-induced arrhythmias and myocardial infarct size in anesthetized albino rabbits. Arrhythmias were induced by reperfusion following a 20 min ligation of the left main coronary artery with a releaseable silk ligature. Early intervention by intravenous infusion of nicorandil (100 microg/kg bolus+10 microg/kg/min) or aprikalim (10 microg/kg bolus+0.1 microg/kg/min) just before and during ischemia increased survival rate (86% and 75% vs. 55% in the control group), significantly decreased the incidence and severity of life-threatening arrhythmias and myocardial infarct size. The antiarrhythmic and cardioprotective effects of both nicorandil and aprikalim were abolished by pretreating the rabbits with 5-hydroxydecanoate (5 mg/kg, i.v. bolus). In conclusion, intervention by intravenous administration of nicorandil and aprikalim (through the selective activation of mitochondrial K(ATP) channels) increased survival rate and exhibited antiarrhythmic and cardioprotective effects during coronary occlusion and reperfusion in anesthetized rabbits when administered prior to and during coronary occlusion. PMID- 11890906 TI - Structure-activity relationship studies with (+/-)-nantenine derivatives for alpha1-adrenoceptor antagonist activity. AB - A series of (+/-)-nantenine derivatives of the natural aporphine alkaloids was synthesized and examined for a blocking action on alpha1-adrenoceptors in rat aorta and A10-cells. The potency of these derivatives was compared with that of an aporphine-related compounds (+)-boldine, an alpha1-adrenoceptor antagonist. Among nine (+/-)-nantenine derivatives having different substituents at N-6, C-1, or C-4 of the aporphine skeleton, (+/-)-domesticine had the most powerful alpha1 adrenoceptor-blocking action. The order of pA2 values was (+/-)-domesticine (8.06+/-0.06)>(+/-)-nordomesticine (7.34+/-0.03)>(+/-)-nantenine (7.03+/ 0.03)>(+)-boldine (6.91+/-0.02)>other derivatives. Study of the structure activity relationships showed that the replacement of a methoxy moiety at C-1 position of (plus minus)-nantenine with a hydroxyl group increased affinity for the receptor. In contrast, replacement of a methyl group with a hydrogen atom or an ethyl group at N-6 position in the (+/-)-nantenine structure decreased affinity for the receptor. These results suggest that a hydroxyl group at the C-1 position and a methyl group at the N-6 position in the (+/-)-nantenine structure are essential for the enhancement of affinity for the alpha1-adrenoceptor. PMID- 11890907 TI - Expression of diamine oxidase (histaminase) in guinea-pig tissues. AB - The expression of mRNA for diamine oxidase (histaminase) and the enzyme activity in guinea-pig tissues were investigated. Reverse transcription-polymerase chain reaction analysis revealed that the message corresponding to the long form present in humans and rats was expressed abundantly in the small intestine and liver. Small but detectable amounts of diamine oxidase mRNA were observed in the kidney, stomach, cerebellum, thalamus+hypothalamus, and cerebral cortex. Northern blot analysis showed that the message (2.8 kb in size) was observed abundantly in the liver and small intestine and was detectable in the kidney and stomach but not in the brain or lung. In situ hybridization showed that diamine oxidase mRNA was localized throughout the liver and epithelial cells of the small intestine. Diamine oxidase activity was detected at various levels in different tissues of the guinea-pig at the following relative abundance: liver>small intestine>lung, kidney>stomach. Histamine dose-dependently induced the contraction of sections of the guinea-pig small intestine, and the pretreatment of the tissue section with aminoguanidine (100 microM), a diamine oxidase inhibitor, but not with S-[4-(N,N dimethylamino)butyl]isothiourea (100 microM), an inhibitor of histamine N methyltransferase, shifted the dose-response curve of histamine-induced contraction to lower concentrations. These results suggest that diamine oxidase has a crucial role in the degradation of histamine in the guinea-pig small intestine and probably in the liver. PMID- 11890909 TI - Whiplash disorders--a review. PMID- 11890908 TI - Effect of topical immunomodulators on acute allergic inflammation and bronchial hyperresponsiveness in sensitised rats. AB - We examined the effects of different immunomodulators administered topically on asthmatic responses in a rat model of asthma. Sensitised Brown-Norway rats were administered rapamycin, SAR943 (32-deoxorapamycin), IMM125 (a hydroxyethyl derivative of D-serine(8)-cyclosporine), and budesonide by intratracheal instillation 1 h prior to allergen challenge. Allergen exposure induced bronchial hyperresponsiveness, accumulation of inflammatory cells in bronchoalveolar lavage fluid, and also an increase in eosinophils and CD2+, CD4+ and CD8+ T cells in the airways. Interleukin-2, interleukin-4, interleukin-5, interleukin-10, and interferon-gamma mRNA expression was upregulated by allergen exposure. Budesonide abolished airway inflammation, suppressed the mRNA expression for interleukin-2, interleukin-4, and interleukin-5 (P<0.03), and bronchial hyperresponsiveness (P<0.05). IMM125 suppressed airway infiltration of eosinophils, and CD8+ T cells (P<0.02), and prevented the upregulated mRNA expression for interleukin-4, interleukin-5, and interferon-gamma (P<0.02). Rapamycin suppressed CD8+ T cell infiltration in airway submucosa (P<0.03), and mRNA expression for interleukin-2 (p<0.002), while SAR943 suppressed interleukin-2, interleukin-4, and interferon gamma mRNA (P<0.05). IMM125, rapamycin and SAR943 did not alter airway submucosal CD2+ and CD4+ T cell infiltration, and bronchial hyperresponsiveness. CD8+ T cells, in contrast to CD4+ T cells, are more susceptible to the inhibition by IMM125 and rapamycin, which also caused greater suppression of Th1 compared to Th2 cytokine mRNA expression. In this acute model of allergic inflammation, differential modulation of Th1 and Th2 cytokines may determine the effects of various immunomodulators on airway inflammation and bronchial hyperresponsiveness. PMID- 11890910 TI - Does prehospital fluid administration impact core body temperature and coagulation functions in combat casualties? AB - BACKGROUND: Administration of large amounts of fluids to trauma patients, in the absence of surgical control, may increase bleeding, cause hypothermia and coagulopathy which may worsen the bleeding and increase morbidity and mortality. The purpose of our study is to examine the impact of prehospital fluid administration to military combat casualties on core body temperature and coagulation functions. METHODS: Prospective data were collected on all cases of moderately (9 < or = ISS < or = 14) and severely (ISS > or = 16) injured victims wounded in South Lebanon, treated by Israeli military physicians and evacuated to hospitals in Israel, over a two-year period. Data regarding prehospital phase of injury (timetables, amount of fluids) and upon hospital arrival (initial core body temperature, prothrombin time [PT], partial thromboplastin time [PTT]) were examined for monotonic relation using Spearman's non-parametric test. RESULTS: Fifty-three moderately injured and 31 severely injured patients were included in the study. The average evacuation time for the moderately injured group was 109.3 +/- 44.8 min, and for the severely injured 100.3 +/- 38.4 min (P value=NS). The mean volume of fluids administered was 2.39 +/- 1.52 and 2.49 +/- 1.47 l, respectively (P=NS). No statistical correlation was found between core body temperature, PT or PTT, measured upon hospital arrival, and prehospital fluid treatment. In addition, no correlation was found between core body temperature on hospital arrival and prehospital time, or between prehospital fluid volumes and prehospital time. The mean core body temperature of the moderately injured patients was 36.8 degrees C, and that of severely injured was 35.8 degrees C (P=0.026). CONCLUSIONS: With proper control of blood loss and avoidance of excessive fluid administration, moderately and severely injured combat casualties in 'low intensity conflict' in South Lebanon can be resuscitated with fluid volumes that do not result in a coagulation deficit or hypothermia. The core body temperature on arrival at the hospital is related to the severity of the injury. PMID- 11890911 TI - Musculoskeletal recovery 5 years after severe injury: long term problems are common. AB - Five years after severe injury (ISS>15), usually involving several body regions, 158 patients were assessed regarding their musculo-skeletal recovery. An earlier paper in this journal about this study 'Injury 29 (1998) 55' showed that when considering the main body regions causing long term disability, 45% were due to bony injuries to the extremities, pelvis and shoulder girdle. We analysed these body areas regarding the degrees of disability and pain and also for problems with activities of daily living, work, sport and mobility. All patients with unstable pelvic fractures had moderate or severe persisting disability and chronic pain. Functional problems with activities of daily living, work, sport and mobility were reported in 28, 86, 100 and 100% of patients, respectively. Patients with stable pelvic fractures had persisting disability in 54% of cases, which was mild in 42% and moderate or severe in 12% of patients. In patients with stable pelvic fractures 54% had chronic pain, which was mild in 24% of patients and moderate or severe in 30% of patients. Functional problems with mobility, work and sport were reported in 38, 19 and 19% of patients, respectively. Patients with shoulder girdle injuries had persisting disability in 48% of cases which was mild in 24% and moderate or severe in 24% of patients. In patients with shoulder girdle injuries 45% had chronic pain, which was mild in 14% and moderate or severe in 31% of patients. Functional problems with activities of daily living, work, sport and mobility were reported in 38, 28, 38 and 38% of patients respectively. Patients with upper limb fractures had persisting disability in 66% of cases which was mild in 34% of patients and moderate or severe in 32% of patients. Chronic pain was present in 62% of these cases, which was mild in 32% and moderate or severe in 34% of patients. Functional problems with activities of daily living, work, sport and mobility were reported in 31, 45, 48 and 66% of patients, respectively. Patients with lower limb fractures had persisting disability in 84% of cases, which was mild in 16% and moderate or severe in 68% of patients. Chronic pain was present in 80% of these cases, which was mild in 24% and moderate or severe in 56% of patients. Functional problems with activities of daily living, work, sport and mobility were reported in 40, 56, 64 and 76% of patients, respectively. Patients with multiple extremity injuries or combinations of pelvic and lower extremity or shoulder girdle and upper extremity injuries were much more likely to have continuing disability compared with those sustaining single bone injuries of that limb. This high disability rate reflecting treatment in 1989-1990, raises the question of whether our present policy of earlier and better fixation and rehabilitation of fractures in severely injured patients (ISS>15) can improve these results. PMID- 11890912 TI - The impact of lower leg compartment syndrome on health related quality of life. AB - Although the aetiology, pathophysiology and treatment of acute compartment syndrome have been well described in the literature, there is limited information on the long-term impact of compartment syndrome on quality of life. We reviewed the medical records and radiographs of all the patients treated with surgical decompression of compartment syndrome. Between 1993 and 1998, 42 cases were identified. There were 30 cases of tibial compartment syndrome and 12 cases involving other limbs. These 30 patients were recalled for a follow-up assessment during which they were asked to complete an EQ-5D (EuroQol), a standardised measure of health related quality of life based on five dimensions (self-care, pain/discomfort, mobility, usual activities and anxiety/depression). Patients were compared with EQ-5D age/sex norms derived from a randomly selected group of patients that had sustained isolated closed tibial shaft fractures. The minimum follow-up time was 12 months. Patients who stated that the appearance of the surgical site was a problem, reported significantly poorer health related quality of life than did patients who had no problem with the appearance. Patients with skin graft reported more problems with pain and discomfort than patients without skin graft. Patients with faster closure times of the wound showed significantly better self-rated health status than patients in whom the wound closure time was longer. Although the patients in this study reported significantly more problems on the dimensions of EQ-5D than were reported in the control group, their overall self-rated health was not statistically different. This study has demonstrated that compartment syndrome may be associated with long-term impact on health related quality of life. PMID- 11890913 TI - Current management of U-shaped sacral fractures or spino-pelvic dissociation. AB - We present a series of four U-shaped sacral fractures, a fracture pattern that is poorly recognised and not included in the standard classifications of these fractures. These fractures are significant as they represent spino-pelvic dissociation, have a high incidence of neurological complications and information regarding current treatment options is sparse. Our four patients underwent early operative stabilisation. Three had ilio-sacral screws inserted, supplemented in one with instrumentation from the lumbar spine to the iliac crest following sacral laminectomy. The other patient had sacral laminectomy with bony stabilisation by instrumentation from the lumbar spine to the iliac crest without ilio-sacral screw fixation. No complications were encountered as a result of fixation and it contributed to the prompt skeletal stabilisation of polytraumatised patients allowing early mobilisation. PMID- 11890914 TI - Treatment of distal femoral nonunions by external fixation with simultaneous length and alignment correction. AB - The use of external fixation for management of distal femoral nonunions may minimise some of the problems frequently encountered in these patients. Fifteen patients treated by external fixation for distal femoral nonunions between 1987 and 1997 were reviewed. There were nine males and six females. The average age was 35.4 years (17-53) with an average follow up of 4.6 years (2-8). Nine followed an open fracture, five a closed fracture and one a femoral osteotomy. Five of the cases were infected nonunions. In all cases an internal fixation device was used as the initial method of treatment. All patients had the nonunion site stabilised with an external fixator. In 12 cases the knee joint was crossed with the fixator to further stabilise the fracture site. All patients had some degree of leg length discrepancy or malalignment that required correction. Fourteen cases united. The other patient united following intramedullary nailing. The average time to union was 10.4 months (4-24). The average range of movement was 80 degrees after treatment. Up to 9 cm of lengthening was achieved using the external fixation system (mean 5.0 cm). The mean angular correction was 15 degrees. One patient had persistent pain despite union at the time of the last follow up. The advantages of preservation of soft tissue, immobilisation of the fracture site by crossing the knee joint and the facility for proximal lengthening make external fixation a definite option in the management of distal femoral nonunions. PMID- 11890915 TI - Clavicular fracture non-union surgical outcome and complications. AB - Symptomatic clavicular fracture non-union is rare. When it does occur, however, it may pose a difficult problem causing pain and functional impairment. The emphasis of this paper is on preopertive disabilities and the postoperative outcome and complications. Twenty patients with clavicular non-union treated operatively from 1989 to 1997 were reviewed, the average follow up was thirty four months. Eleven fractures were midshaft, eight were lateral third and one medial third. A detailed proforma was completed with the patients documenting preoperative symptoms, outcome after surgery and complications. A literature search was carried out to find out the incidence of complications related to operating on post-traumatic clavicular non-union. All of the twenty fractures non unions duly united after surgical intervention although three required early adjustment or change of metal work. The subjective and objective outcomes were good in 19 cases and poor in one. The postoperative complications included three implant failures, one stiff shoulder, two patients with numbness below the scar and one patient with an infected donor site wound. The literature search revealed that from 24 publications and 301 patients who had operations for clavicular non union ther were 18 (6%) reported complications related to metal work, 45 (15%) reported complications related to soft tissues, seven (2%) complications related to the scar and 24 (8%) failures of union. Symptomatic clavicular non-union can cause severe disabilities. Good outcome, at low risk, can be expected from internal fixation and bone grafting of midshaft non-unions. Although there are only eight cases of lateral third clavicular non-unions, this is the largest series in the literature. Furthermore, the study clearly demonstrates both the difficulties treating this fracture surgically and a procedure to be avoided (acromio-clavicular bridging), which, again has not been previously addressed. PMID- 11890916 TI - The vascularity of atrophic non-unions. AB - The objective of this study was to assess the vascularity of atrophic non-unions using an experimental animal model. Twenty skeletally mature female rabbits were randomly divided into control and experimental groups that were killed 1, 8, or 16 weeks after surgery. The experimental groups underwent surgery to induce an atrophic non-union whereas the control groups underwent a similar operation but which resulted in union. Using immunocytochemical techniques blood vessels were identified in histological sections obtained from the osteotomy site. The concentration of the vessels within the osteotomy gaps was measured, as was the serum concentration of an important angiogenic factor: endothelial cell stimulating angiogenesis factor (ESAF). The results demonstrated a significant difference between the control and the experimental groups in the concentration of vessels within the gap at 1 week but there was no significant difference between those groups at 8 weeks. There were no significant differences in the ESAF concentration between the groups at any time points. We concluded that established atrophic non-unions can be well vascularised and that measurements of serum levels of ESAF could not distinguish between those osteotomies that would unite and those that would progress to non-union. PMID- 11890917 TI - Histological assessment of the presence or absence of infection in fracture non union. AB - BACKGROUND: Infection is a major cause of non-unions. Infection is not always evident clinically, nor on bacteriological analysis. If it is untreated, non union treatment may fail. AIM: To establish whether histological analysis is of value as an adjunct to microbiology in establishing the diagnosis of infection. METHODS: Sixty patients who had both bacteriological and histological analysis of their non-unions were studied. Infection was indicated by an acute inflammatory response. In 45 of the 60 fractures, microbiological and clinical diagnoses were in agreement; in this subset the histology results were compared to the established diagnoses. RESULTS: The histological diagnosis for the 45 fractures in the group with a definite diagnosis gave four false negatives but no false positives. This represents a sensitivity of 87.1%, (95% CI, 70-96%) and a 100% specificity (95% CI, 77-100%). The overall accuracy was 91.1%. (95% CI, 79-98%) The predictive value of a positive test was 100% and of a negative test was 77.8%. In 25% of the series a definite diagnosis could not be made with purely clinical and microbiological information; with the additional histological information it was possible to determine the infection status of the non-union. CONCLUSION: Histology is of particular use when the microbiology is negative or equivocal. PMID- 11890918 TI - Contamination of ballistic fractures: an in vitro model. AB - This study allowed the development of an in vitro model of a high-energy ballistic fracture. Direct fractures are more heavily contaminated than indirect fractures, and the spread of contamination is more extensive than examination of the wounds, particularly the entry wounds, would suggest. The major spread of contamination is along tissue planes, and we would recommend that these are thoroughly lavaged in the management of the fracture. The fracture site is heavily contaminated, with endosteal spread in direct fractures. However, there appears to be relatively little bony contamination beyond the fracture site. PMID- 11890919 TI - Supracondylar missile fractures of the femur. AB - We present the results of the surgical treatment of 50 supracondylar missile fractures of the femur in 48 patients wounded in the territory of the former Yugoslavia in the period June 1991-October 1995. The injuries were caused by bullets in 28 (58.3%) patients and by fragments of mines or other explosive devices in 20 (41.7%) patients. In nine (18.0%) missile supracondylar fractures there were associated fractures of articular surfaces in the knee-joint. Neurovascular bundle injuries were present in 19 (38.0%) and vascular injuries alone in 17 (34.0%) limbs. Bone fragments were stabilized by external fixation in 43 (86.0%) and by plaster of Paris in five (10.0%) limbs. Primary reconstruction of large blood vessel was necessary in 16 (32.0%) limbs. In total eight (16.0%) above knee amputations were done. Coverage of soft tissue defects was required in 19 (38.0%) limbs. Early and late postoperative complications occurred in 26 (52.0%) limbs. Additional surgical procedures were necessary in 15 patients. Definitive results showed a large number of limb shortenings, nerve paralyses and contractures of the knee-joint. PMID- 11890921 TI - Snowboarding splenic injury: four case reports. AB - With the rapidly increasing number of snowboarders, the incidence of injuries has recently become higher. From 1994 to 1995, we encountered four snowboarders with splenic injuries in one season. In three of the four patients the splenic injuries were caused by striking the abdomen with their own elbow when falling by themselves, of which emergent splenectomy was required in two patients. In the other one the collision with another snowboarder caused the splenic injury and splenorrhaphy was performed. Because snowboarders have both feet fixed on a board and do not have poles, they are prone to fall on the left upper limb in the proceeding direction, resulting in the striking of the left upper abdomen. Because in snowboarders splenic injury is caused mostly by a blow from their own left elbow at the time of falling, informing the mechanisms of splenic injuries will serve a speedy correct diagnosis for the doctors. PMID- 11890920 TI - Hindfoot injuries due to landmine blast accidents. AB - Landmines were initially developed as anti-tank weapons. They are still used indiscriminately and in a disorganised fashion, violating the United Nations Treaty on their use [United Nations (1980)]. The injury produced by these devices is variable depending upon the construction and strength of the landmine and body parts coming in contact with the landmine at the time of detonation. The purpose of the present study was to report the type of landmine-blast injuries of the lower limbs and the surgical options available to treat them. Twenty-eight patients, all with lower limb injuries were included in the present study. They had received injuries on the control line of the troubled Jammu and Kashmir regions in the north of Pakistan. All were male patients between the age of 13 and 55 years. A salvage procedure for the forefoot was possible on four patients only and all the rest had a below-knee amputation. Time lapsed between the injury and receiving medical help was the crucial determining factor as to the final outcome of the limb. We believe that the pattern of injury, amount of energy dissipation and part of body in contact with the landmine at the time of explosion are the main determining factors for the final outcome. If skin along with the underlying soft tissue and the neuro-vascular structures on the dorsum of the foot are spared then an attempt can be made at limb salvage. PMID- 11890922 TI - Distal humeral epiphysis fracture separation in neonates -- diagnosis using MRI scan. PMID- 11890923 TI - Vertebrobasilar occlusion following snowboarding: a case report and review. PMID- 11890924 TI - True skier's thumb in childhood. PMID- 11890925 TI - A complex thoracic fracture extending from fourth thoracic to twelfth thoracic vertebrae without neurological deficit--a case report. PMID- 11890929 TI - Molecular analysis of mitochondrial DNA mutations from bleomycin-treated rats. AB - In our previous studies, we have shown the mutagenicity of bleomycin (BLM) at the nuclear hprt locus. In the present study we have analyzed mutagenic effects of BLM in mitochondrial DNA (mtDNA) using short extension-PCR (SE-PCR) method for detection of low-copy deletions. Fisher 344 rats were treated with a single dose of BLM and total DNA preparations from splenic lymphocytes were processed in SE PCR assay. Spontaneous deletions were typically flanked by direct repeats (78.5%), while the in BLM-treated group, direct repeats were found in only 46.6% of breakpoints. The ratio between deletions based on direct repeats and random sequence deletions changed from 3.67 in control group to 0.87 in BLM-treated animals, which corresponds to an approximate 1.7-fold increase in the deletion mutation frequency. Furthermore, 62.5% of deletions not flanked by direct repeats in the treated group contained cleavage sites for BLM. The localization of breakpoints was not entirely random. We have found four clusters containing deletions from both groups indicative of deletion hot spots. The results indicate that BLM exposure may be associated with the induction of mtDNA mutations, and suggest the utility of SE-PCR method for evaluating drug-induced genotoxicity. PMID- 11890930 TI - X-ray induced mutation in Syrian hamster fetal cells. AB - Transabdominal X-rays are a risk factor for childhood leukemia, and X-ray exposure of mouse fetuses has led to increases in both mutations and initiated tumors in offspring. However, fetal sensitivity and dose-response characteristics with regard to transplacental mutagenesis by X-rays have never been quantified. In the current experiment, pregnant Syrian hamsters at day 12 of gestation were irradiated with 300-kV X-rays. Twenty-four hours later, the fetuses were removed and their cells were allowed a 5 day expression time in culture. They were then seeded for colony formation and also for mutation selection by 6-thioguanine (6 TG). Mutation frequency was linear over the entire dose range, 10-600 R. The average induced 6-TG mutant frequency was 4.7 x 10(-7) per R. These results suggest that fetal cells are highly sensitive to induction of mutations by X rays, and that a no-effect threshold is not likely. The 10 R dose caused a 25 fold increase in mutation frequency over the historical control, 45 x 10(-7) versus 1.8 x 10(-7), an increase per R of 2.5-fold. Increased risk of childhood cancer related to obstetrical transabdominal X-ray has also been estimated at 2.5 fold per R. Thus, our results are consistent with mutation contributing to this effect. PMID- 11890931 TI - SN2 DNA-alkylating agent-induced phosphorylation of p53 and activation of p21 gene expression. AB - p53 is an important player in the cellular response to genotoxic stress whose functions are regulated by phosphorylation of a number of serine and threonine residues. Phosphorylation of p53 influences its DNA-binding and gene regulation activities. This study examines p53 phosphorylation in HCT-116 (MMR-deficient) and HCT-116+ch3 (MMR-proficient) human colon cancer cells treated with a S(N)2 DNA-alkylating agent, methylmethane sulfonate (MMS). MMS induces phosphorylation of p53 on Ser15 and Ser392 in a dose- and time-dependent manner. MMS-induced p53 phosphorylation is independent of DNA mismatch repair (MMR) activity. Nuclear extracts from MMS-treated HCT-116 cells had higher p21WAF1/Cip1 (p21) promoter DNA-binding activity in vitro opposed to untreated cells. After MMS treatment, the activation of the cloned p21 promoter in a transient transfection assay and endogenous p21 mRNA levels in HCT-116(p53+/+) versus HCT-116(p53-/-) cells increased, which correlates with an increased levels of phospho-p53(Ser15) and phospho-p53(Ser392). These results suggest that SN2 DNA-alkylating agent-induced phosphorylation of p53 on Ser15 and Ser392 increases its DNA-binding properties to cause an increased expression of p21 that may play a role in cell cycle arrest and/or apoptosis of HCT-116 cells. PMID- 11890932 TI - Effects of dietary antioxidants on DNA damage in lysed cells using a modified comet assay procedure. AB - A modified version of the comet assay was employed to investigate the effect in vitro of dietary antioxidants in the subcellular environment. Human lymphocytes were isolated, embedded in agarose gel, lysed in high ionic strength solution with Triton X-100, and then incubated for 30 min with antioxidants at different concentrations. Gels were washed, and the comet assay performed on cells stressed by 5 min incubation with 45 microM hydrogen peroxide and on unstressed cells in parallel. Results showed that alpha-tocopherol was protective against oxidant stress, whereas caffeic acid did not protect, and at high concentration (100 microM) caused increased DNA damage. Results for quercetin suggested a direct damaging effect, but this did not reach statistical significance. However, at low concentration (3.1 microM), quercetin appeared protective. Thus some dietary antioxidants that have been shown previously to have a protective effect in the 'standard', whole-cell, comet assay cause DNA damage in this lysed-cell version. The cell membrane may have an important role in limiting cellular access of these 'double-edged' antioxidants. Furthermore, the absolute concentration and the presence of complementary or synergistic intracellular antioxidants may delineate the type of action of a putative antioxidant. We suggest that, used in conjunction with the standard comet assay, this lysed-cell version is useful for assessing the effect of the cell membrane and intracellular systems on susceptibility of DNA to oxidative damage, and will help determine the mechanism of protection or damage by phytochemicals. PMID- 11890933 TI - Genotoxic and recombinogenic activities of the two beta-carboline alkaloids harman and harmine in Saccharomyces cerevisiae. AB - The cytotoxical beta-carboline alkaloids harman and harmine occur in medical plants and in a variety of foods, alcoholic beverages, and industrial waste. We applied them to the yeast Saccharomyces cerevisiae to test for putative genotoxicity, mutagenicity and recombinogenicity and to determine whether harman and harmine produced repairable DNA damage. Harmine was more cytotoxic than harman for exponentially growing haploid and diploid cells. Only harmine-induced crossing-over and mitotic gene conversion but both alkaloids were frameshift mutagens in yeast. Mutants defective in excision-resynthesis repair (rad3 and rad1), in error-prone repair (rad6) and in recombinational repair (rad52) showed enhanced sensitivity to harmine and harman, but the ranking of sensitivities was different for the two alkaloids. It appears that both alkaloids are probably capable of inducing DNA single and/or double strand breaks. An epistatic interaction was shown between rad3-e5 and rad52-1 mutants alleles, indicating that excision-resynthesis and strand-break repair may have common steps in the repair of DNA damage induced by these alkaloids. The non-epistatic interaction observed in rad1Delta rad6Delta double mutants indicated that both excision resynthesis and error-prone repair are independently involved in repair of harman and harmine-induced DNA lesions. PMID- 11890934 TI - Mechanism of peroxidase-mediated oxidation of carcinogenic o-anisidine and its binding to DNA. AB - 2-Methoxyaniline (o-anisidine) is a urinary bladder carcinogen in both mice and rats. Since the urinary bladder contains substantial peroxidase activity, we investigated the metabolism of this carcinogen by prostaglandin H synthase (PHS), a prominent enzyme in the urinary bladder, and lactoperoxidase as model mammalian peroxidases. Horseradish peroxidase (HRP)-mediated oxidation of o-anisidine was also determined and compared with the reactions catalyzed by mammalian peroxidases. All three peroxidases oxidized o-anisidine via a radical mechanism. Using HPLC combined with electrospray tandem mass spectrometry, we determined that peroxidases oxidized o-anisidine to a diimine metabolite, which subsequently hydrolyzed to form a quinone imine. Two additional metabolites were identified as a dimer linked by an azo bond and another metabolite consisting of three methoxybenzene rings, which exact structure has not been identified as yet. Using [14C]-labeled o-anisidine, we observed substantial peroxidase-dependent covalent binding of o-anisidine to DNA, tRNA and polydeoxynucleotides [poly(dX)]. The 32P postlabeling assay (a standard procedure and enrichment of adducts by digestion with nuclease P1 or by extraction into 1-butanol prior to 32P-labeling) was employed as the second method to detect and quantitate binding of o-anisidine to DNA. Using these versions of the 32P-postlabeling technique we did not observe any DNA adducts derived from o-anisidine. The o-anisidine-DNA adducts became detectable only when DNA modified by o-anisidine was digested using three times higher concentrations of micrococcal nuclease and spleen phosphodiesterase (MN/SPD). We found deoxyguanosine to be the target for o-anisidine binding in DNA using poly(dX) and deoxyguanosine 3'-monophosphate (dGp). A diimine metabolite of o-anisidine is the reactive species forming adducts in dGp. The results strongly indicate that peroxidases play an important role in o-anisidine metabolism to reactive species, which might be responsible for its genotoxicity, and its carcinogenicity to the urinary bladder in rodents. The limitation of the 32P postlabeling technique to analyze DNA adducts derived from o-anisidine as a means to estimate its genotoxicity is discussed. PMID- 11890936 TI - DNA adducts and liver DNA replication in rats during chronic exposure to N nitrosodimethylamine (NDMA) and their relationships to the dose-dependence of NDMA hepatocarcinogenesis. AB - Exposure of rats to the hepatocarcinogen N-nitrosodimethylamine (NDMA) (0.2-2.64 ppm in the drinking water) for up to 180 days resulted in rapid accumulation of N7- and O6-methylguanine in liver and white blood cell DNA, maximum adduct levels being reached within 1-7 days, depending on the dose. The levels of both adducts remained constant up to treatment day 28, subsequently declining slowly to about 40% of maximal levels for the liver and 60% for white blood cells by day 180. In order to elucidate the role of DNA replication in NDMA hepatocarcinogenesis, changes in liver cell labeling index (LI) were also measured on treatment days 21, 120 and 180. Although the time- and dose-dependence of the observed effects were complex, a clear trend towards increased rates of hepatocyte LI, as indicated by BrdU incorporation, with increasing NDMA doses was evident, particularly above 1 ppm, a concentration above which NDMA hepatocarcinogenicity is known to increase sharply. In contrast, no increase in Kupffer cell DNA replication was found at any of the doses employed, in accordance with the low susceptibility of these cells to NDMA-induced carcinogenesis. No significant increase in the occurrence of necrotic or apoptotic cells was noted under the treatment conditions employed. These results suggest that, in addition to the accumulation of DNA damage, alterations in hepatocyte DNA replication during the chronic NDMA exposure may influence the dose-dependence of its carcinogenic efficacy. PMID- 11890935 TI - Thymic lymphomas arising in Msh2 deficient mice display a large increase in mutation frequency and an altered mutational spectrum. AB - Mismatch repair (MMR) genes, such as Msh2, are classified as "mutator" genes, responsible for the microsatellite instability identified in many tumors. In the current study, the mutation frequency and mutational spectrum in thymic lymphoma arising in Msh2 deficient mice are investigated. Thymic lymphoma developed in Msh2-/- background displayed an eight to nine-fold increase in mutation frequency compared to the normal thymi in Msh2 deficient animals. Sequencing demonstrated significantly different mutational spectra between normal thymus tissue and thymic lymphomas in Msh2-/- mice (P=0.02). The tumor mutational spectrum is characterized by an increase in base substitutions occurring at A:T sites, and multiple mutations, as well as a minor increase in -1 frameshifts. We analyzed mutations in different parts of the tumors, and different regional hotspots could be identified. Several hotspot mutations that are a rare event in normal tissues were identified in the tumor tissues. We conclude that thymic lymphomas arising in Msh2 deficient genetic background are hypermutable and the altered mutational spectrum might be an indication of infidelity of DNA replication during tumorigenesis. PMID- 11890937 TI - Mutagen sensitivity of peripheral blood from women carrying a BRCA1 or BRCA2 mutation. AB - We are studying the induction and repair of DNA damage in lymphocytes of women from families with familial breast cancer and a heterozygous mutation in the breast cancer susceptibility genes BRCA1 or BRCA2. Besides various other functions, BRCA proteins seem to be involved in DNA repair processes like transcription-coupled and double-strand break (dsb) repair. Our previous results indicated a close relationship between the presence of a BRCA1 mutation and sensitivity for the induction of micronuclei (MN) by gamma irradiation and hydrogen peroxide (H2O2). In contrast to the results with the micronucleus assay, we found no significant individual difference between women with and without a BRCA1 mutation with respect to the induction and repair of DNA damage in the alkaline comet assay. We now investigated further cases heterozygous for a BRCA1 mutation and cases heterozygous for a BRCA2 mutation and show that enhanced micronucleus formation after gamma irradiation and H2O2-treatment is also a feature of lymphocytes carrying a BRCA2 mutation. Investigations with the comet assay did not reveal clear differences with regard to the induction of DNA damage on the individual level. There were also no significant differences between blood samples carrying a BRCA1 or BRCA2 mutation and blood samples from normal controls when the repair capacities (i.e. the kinetics of the removal of radiation-induced DNA effects in the comet assay) were compared. Our results indicate that mutagen sensitivity of lymphocytes heterozygous for a BRCA2 mutation is similar to that of cells with a BRCA1 mutation and BRCA1 and BRCA2 cannot be differentiated at present with the micronucleus test (MNT) or the comet assay. PMID- 11890938 TI - Mutagenic profiles of carbazole in the male germ cells of Swiss albino mice. AB - Mutagenic effect of carbazole was evaluated by employing dominant lethal mutation and sperm head abnormality assays in male Swiss albino mice. For the dominant lethal mutation assay, adult male mice were treated for five consecutive days either with 30 or 60 mg/kg body weight (b.w.) of carbazole by single intraperitoneal (i.p.) injection. For the sperm head abnormality assay mice were treated with 50, 100, 150, 200 and 300 mg/kg b.w as a single i.p. injection. Treatment of adult male mice with carbazole resulted in induction of dominant lethal mutation and abnormal sperm heads. The results show that carbazole is mutagenic in male germ cells of mice. PMID- 11890939 TI - Mutation analysis of the human CYP3A4 gene 5' regulatory region: population screening using non-radioactive SSCP. AB - Human CYP3A4 is the major cytochrome P450 isoenzyme in adult human liver and is known to metabolise many xenobiotic and endogenous compounds. There is substantial inter-individual variation in the hepatic levels of CYP3A4. Although, polymorphic mutations have been reported in the 5' regulatory region of the CYP3A4 gene, those that have been investigated so far do not appear to have any effect on gene expression. To determine whether other mutations exist in this region of the gene, we have performed a new population screen on a panel of 101 human DNA samples. A 1140 bp section of the 5' proximal regulatory region of the CYP3A4 gene, containing numerous regulatory motifs, was amplified from genomic DNA as three overlapping segments. The 300 bp distal enhancer region at -7.9kb containing additional regulatory motifs was also amplified. Mutation analysis of the resulting PCR products was carried out using non-radioactive single strand conformation polymorphism (SSCP) and confirmatory sequencing of both DNA strands in those samples showing extra SSCP bands. In addition to detection of the previously reported CYP3A4*1B allele in nine subjects, three novel alleles were found: CYP3A4*1E (having a T-->A transversion at -369 in one subject), CYP3A4*1F (having a C-->G tranversion at -747 in 17 subjects) and CYP3A4*15B containing a nine-nucleotide insertion between -845 and -844 linked to an A-->G transition at 392 and a G-->A transition in exon 6 (position 485 in the cDNA) in one subject. All the novel alleles were heterozygous. No mutations were found in the upstream distal enhancer region. Our results clearly indicate that this rapid and simple SSCP approach can reveal mutant alleles in drug metabolising enzyme genes. Detection and determination of the frequency of novel alleles in CYP3A4 will assist investigation of the relationship between genotype, xenobiotic metabolism and toxicity in the CYP3A family of isoenzymes. PMID- 11890940 TI - DNA adducts of styrene-7,8-oxide in target and non-target organs for tumor induction in rat and mouse after repeated inhalation exposure to styrene. AB - Styrene by inhalation had been shown to increase the lung tumor incidence in mice at 20 ppm and higher, but was not carcinogenic in rats at up to 1000 ppm. Styrene 7,8-oxide, the major metabolic intermediate, has weak electrophilic reactivity. Therefore, DNA adduct formation was expected at a low level and a 32P postlabeling method for a determination of the two regioisomeric 2'-deoxyguanosyl O6-adducts at the alpha(7)- and beta(8)-positions had been established. The first question was whether DNA adducts could be measured in the rat at the end of the 2 years exposure of a bioassay for carcinogenicity, even though tumor incidence was not increased. Liver samples of male and female CD rats were available for DNA adduct analysis. Adducts were above the limit of detection only in the highest dose group (1000 ppm), with median levels of 9 and 8 adducts per 10(7) nucleotides in males and females, respectively (sum of alpha- and beta-adducts). The result indicates that the rat liver tolerated a relatively high steady-state level of styrene-induced DNA adducts without detectable increase in tumor formation. The second question was whether different DNA adduct levels in the lung of rats and mice could account for the species difference in tumor incidence. Groups of female CD-1 mice were exposed for 2 weeks to 0, 40, and 160 ppm styrene (6h per day; 5 days per week), female CD rats were exposed to 0 and 500 ppm. In none of the lung DNA samples were adducts above a limit of detection of 1 adduct per 10(7) DNA nucleotides. The data indicate that species- and organ specific tumor induction by styrene is not reflected by DNA adduct levels determined in tissue homogenate. The particular susceptibility of the mouse lung might have to be based on other reactive metabolites and DNA adducts, indirect DNA damage and/or cell-type specific toxicity and tumor promotion. PMID- 11890941 TI - Dose-dependent increase or decrease of somatic intrachromosomal recombination produced by etoposide. AB - Chromosomal inversions and deletions can occur via somatic intrachromosomal recombination (SICR), a mechanism known to be important in mutagenesis and carcinogenesis. Here, we demonstrate a dose-dependent increase or decrease in SICR inversion frequency both in vivo and in vitro after treatment with etoposide, using the pKZ1 mouse mutagenesis model. pKZ1 mice received a single intraperitoneal injection of etoposide dose ranging from 0.0005 to 50mg/kg. Animals were sacrificed 3 days after treatment and the spleen was analysed for SICR. A significant 1.4-3.1-fold induction of SICR inversion events was detected in pKZ1 mice after treatment with etoposide doses ranging from 0.05 to 50 mg/kg etoposide. However, inversion frequencies after treatment with 0.0005 and 0.005 mg/kg etoposide decreased significantly to 0.67 and 0.43 of the levels observed in control animals, respectively. A pKZ1 mouse hybridoma cell line was exposed to etoposide (1-1000 nM) and a similar pattern of SICR response to that detected in vivo was observed. A significant 2.3-4.6-fold induction of SICR inversions was observed in pKZ1 cells treated with 100 and 1000 nM etoposide. Inversion frequencies after treatment with 1 and 10nM etoposide decreased significantly to 0.31 and 0.5 of the level observed in control cell lines. Our in vitro studies complement our in vivo studies and exclude a kinetic phenomenon as the responsible mechanism of reduction in SICR in response to low dose etoposide. Determination of the exact mechanism and significance of recombination suppression at low doses of etoposide treatment requires further investigation. PMID- 11890942 TI - Chlorambucil-induced high mutation rate and suicidal gene downregulation in a base excision repair-deficient Escherichia coli strain. AB - Chlorambucil (CLB; N,N-bis(2-chloroethyl)-p-aminophenylbutyric acid) is a bifunctional alkylating agent widely used as an anticancer drug and also as an immunosuppressant. Its chemical structure and clinical experience indicate that CLB is mutagenic and carcinogenic. We have investigated the ability of CLB to induce mutations and gene expression changes in the wild-type (WT) Escherichia coli strain AB1157 and in the base excision repair-deficient (alkA1, tag-1) E. coli strain MV1932 using a rifampicin (rif) forward mutation system and a cDNA array method. The results showed that CLB is a potent mutagen in MV1932 cells compared with the E. coli WT strain AB1157, emphasizing the role of 3 methyladenine DNA glycosylases I and II in protecting the cells from CLB-induced DNA damage and subsequent mutations. Global gene expression profiling revealed that nine genes in WT E. coli and 100 genes in MV1932, of a total of 4290 genes, responded at least 2.5-fold to CLB. Interestingly, all of these MV1932 genes were downregulated, while 22% were upregulated in WT cells. The downregulated genes in MV1932 represented most (19/23) functional categories, and unexpectedly, many of them code for proteins responsible for genomic integrity. These include: (i) RecF (SOS-response, adaptive mutation), (ii) RecC (resistance to cross-linking agents), (iii) HepA (DNA repair, a possible substitute of RecBCD), (iv) Ssb (DNA recombination repair, controls RecBCD), and (v) SbcC (genetic recombination). Our results strongly suggest that in addition to the DNA damage itself, the downregulation of central protecting genes is responsible for the decreased cell survival (demonstrated in a previous work) and the increased mutation rate (this work) of DNA repair-deficient cells, when exposed to CLB. PMID- 11890943 TI - Attenuation of DNA polymerase beta-dependent base excision repair and increased DMS-induced mutagenicity in aged mice. AB - The biological mechanisms responsible for aging remain poorly understood. We propose that increases in DNA damage and mutations that occur with age result from a reduced ability to repair DNA damage. To test this hypothesis, we have measured the ability to repair DNA damage in vitro by the base excision repair (BER) pathway in tissues of young (4-month-old) and old (24-month-old) C57BL/6 mice. We find in all tissues tested (brain, liver, spleen and testes), the ability to repair damage is significantly reduced (50-75%; P<0.01) with age, and that the reduction in repair capacity seen with age correlates with decreased levels of DNA polymerase beta (beta-pol) enzymatic activity, protein and mRNA. To determine the biological relevance of this age-related decline in BER, we measured spontaneous and chemically induced lacI mutation frequency in young and old animals. In line with previous findings, we observed a three-fold increase in spontaneous mutation frequency in aged animals. Interestingly, lacI mutation frequency in response to dimethyl sulfate (DMS) does not significantly increase in young animals whereas identical exposure in aged animals results in a five fold increase in mutation frequency. Because DMS induces DNA damage processed by the BER pathway, it is suggested that the increased mutagenicity of DMS with age is related to the decline in BER capacity that occurs with age. The inability of the BER pathway to repair damages that accumulate with age may provide a mechanistic explanation for the well-established phenotype of DNA damage accumulation with age. PMID- 11890944 TI - A novel single molecule analysis of spontaneous and radiation-induced mutation at a mouse tandem repeat locus. AB - Expanded simple tandem repeat (ESTR) loci include some of the most unstable DNA in the mouse genome and have been extensively used in pedigree studies of germline mutation. We now show that repeat DNA instability at the mouse ESTR locus Ms6-hm can also be monitored by single molecule PCR analysis of genomic DNA. Unlike unstable human minisatellites which mutate almost exclusively in the germline by a meiotic recombination-based process, mouse Ms6-hm shows repeat instability both in germinal (sperm) DNA and in somatic (spleen, brain) DNA. There is no significant variation in mutation frequency between mice of the same inbred strain. However, significant variation occurs between tissues, with mice showing the highest mutation frequency in sperm. The size spectra of somatic and sperm mutants are indistinguishable and heavily biased towards gains and losses of only a few repeat units, suggesting repeat turnover by a mitotic replication slippage process operating both in the soma and in the germline. Analysis of male mice following acute pre-meiotic exposure to X-rays showed a significant increase in sperm but not somatic mutation frequency, though no change in the size spectrum of mutants. The level of radiation-induced mutation at Ms6-hm was indistinguishable from that established by conventional pedigree analysis following paternal irradiation. This confirms that mouse ESTR loci are very sensitive to ionizing radiation and establishes that induced germline mutation results from radiation-induced mutant alleles being present in sperm, rather than from unrepaired sperm DNA lesions that subsequently lead to the appearance of mutants in the early embryo. This single molecule monitoring system has the potential to substantially reduce the number of mice needed for germline mutation monitoring, and can be used to study not only germline mutation but also somatic mutation in vivo and in cell culture. PMID- 11890945 TI - Effect of conjugated linoleic acid on the formation of spontaneous and PhIP induced mutation in the colon and cecum of rats. AB - Conjugated linoleic acid (CLA), a mixture of positional and geometric isomers of linoleic acid, has been reported to inhibit chemically induced mammary and colon carcinogenesis in rodents. In a preliminary experiment, we found that CLA significantly reduced the induction of mutations by the dietary carcinogen 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in the distal colon in male rats. Here, the chemopreventive properties of CLA were further evaluated by assessing its effect on PhIP-induced mutation and aberrant crypt foci (ACF) in both male and female rats. CLA (1%, w/w) was added to the diet (1) from weaning to 50-day-old, or (2) starting 1 week prior to exposure to PhIP. The 50-day-old Big Blue and F344 rats were then exposed to 100 ppm PhIP for 47 days. No sex differences were observed in mutagenic response to the various treatments in either the distal colon or cecum. The mutation frequency (MF) in the cecum and the distal colon from control animals is 4.3+/-1.3 and 5.3+/-1.4 x 10(-5), respectively showing no statistically significant difference. Administration of PhIP induced a four-fold increase in the MF in the cecum and a seven-fold increase in the distal colon compared to the corresponding controls. Supplementation of the diet with CLA lowered the PhIP-induced MF in the distal colon by 23% (P<0.03), but had no effect in the cecum. The PhIP-induced ACF, determined 9 weeks after the termination of treatment with PhIP, were 0.75 ACF/rat, with 1.7 aberrant crypts /ACF in the colon of male rats, all located in the distal colon. This induction was completely inhibited by the addition of CLA. PMID- 11890946 TI - Effect of chronic and acute stress on ectonucleotidase activities in spinal cord. AB - We have previously observed that, while acute stress induces analgesia, chronic stress causes a hyperalgesic response in male rats. No effect was observed in females. There is increasing evidence that both ATP and adenosine can modulate pain. Extracellular ATP and ADP are hydrolyzed by an apyrase in synaptosomes from the peripheral and central nervous systems. In the present study, we investigated the effect of chronic and acute stress on ATPase-ADPase and 5'-nucleotidase activities in spinal cord of male and female rats. Adult male and female Wistar rats were submitted to 1 h restraint stress/day for 1 day (acute) or 40 days (chronic) and were sacrificed 24 h later. ATPase-ADPase activities were assayed in the synaptosomal fraction obtained from the spinal cord of control and stressed animals. ADP hydrolysis was decreased 25% in chronically stressed males, while no change was observed on ATPase activity. There was an increase in the 5' nucleotidase activity in the same group. No effect on ADPase, ATPase or on 5' nucleotidase activity was observed in females with chronic stress, or after acute stress neither in males or females. Chronic stress reduced ADP hydrolysis and increased 5'-nucleotidase activity in the spinal cord in male rats. PMID- 11890947 TI - The control of water and sodium chloride intake by postingestional and orosensory stimulation in water-deprived rats. AB - We investigated the role of oropharyngeal and postingestional stimulation in the control of the intake of water and NaCl solutions by testing water-deprived rats under real- and sham-drinking conditions with water, 0.150, 0.225, and 0.300 M NaCl solutions. A series of real-drinking tests was given until intake stabilized followed by a series of sham-drinking tests with the same solution. When sham intake stabilized the concentration was increased and the series of real- followed by sham-drinking tests was repeated. Intake of water and 0.150 M NaCl in the first sham-drinking test was significantly greater than in the preceding real drinking test and did not change with real- or sham-drinking experience. In contrast, intake of 0.225 and 0.300 M NaCl in the first sham-drinking test was not significantly greater than in the preceding real-drinking test but increased with sham-drinking experience. Real intake of 0.225 and 0.300 M NaCl following sham-drinking experience with a lower concentration declined significantly with real-drinking experience. These results show that postingestional stimulation plays a direct role in the control of the intake of water and isotonic saline with little or no orosensory contribution. In contrast, conditioned orosensory responsiveness played the central role in the control of the intake of the two hypertonic solutions with little or no direct contribution from postingestional stimulation. Postingestional stimulation, however, played an indirect role by serving as the unconditioned stimulus for the conditioned orosensory control. PMID- 11890948 TI - Spatial memory deficit and emotional abnormality in OLETF rats. AB - Cholecystokinin (CCK) is deeply involved in the control of learning and emotional behaviors. The authors characterize the behavioral properties of Otsuka Long Evans Tokushima Fatty (OLETF) rats, which lack the CCK-A receptor because of a genetic abnormality. In the Morris water-maze task, the OLETF rats showed an impaired spatial memory. In the inhibitory avoidance test, they showed facilitating response 24 h after training. Hypoalgesia was observed in a hot plate test. In the elevated plus-maze and food neophobia test, OLETF rats showed an anxiety-like response. In addition, OLETF rats were hypoactive in the Morris water-maze and the elevated plus-maze. The results suggest that the OLETF rats showed a spatial memory deficit, hypoactivity and anxiety due, at least in part, to the lack of CCK-A receptors. PMID- 11890949 TI - Neuroendocrine control of urine-marking behavior in male rats. AB - Sexually experienced Wistar male rats were used to investigate (a) urine voiding in the presence of nearby estrous females and the control of such voiding by (b) steroid hormones and (c) peripheral nerves supplying the genitourinary system. The first experiment showed that males always have a low rate of urine voiding that is significantly increased when a receptive female is around. Thus, it is suggested that an airborne scent from the female stimulates the olfactory system of males, triggering urine emission to transmit sex-related messages, i.e., male rats display the well-known urine-marking behavior of mammals. The number of urine marks and sniffing to females decreased after castration, and were restored after exogenous treatment with testosterone or estradiol. The proposed hypothesis is that airborne scents from the female activate the aromatization process in nuclei of the olfactory pathway of the male, evoking a cascade of neuronal responses that finish in urine marking. Peripheral nerves supplying the genitourinary system are the viscerocutaneous branch of the pelvic nerve (Vc) and the hypogastric (Hg). Data showed that both nerves are important for the central control of urine storage and voiding. Transection of Vc almost blocked urine marking, while Hg lesion increased the number of marks. Thus, it is discussed that Vc is the most important nerve in charge of voiding the bladder, and that Hg is important for continence. PMID- 11890950 TI - Anatomical and physiological characteristics of perineal muscles in the female rabbit. AB - Little information is available on the participation of the perineal striated muscles in female reproductive processes. Here, we describe the gross anatomy and innervation of two striated perineal muscles in the female rabbit, the bulbospongiosus (BSM) and ischiocavernosus (ISM), and analyze their reflex electromyographic (EMG) activity in response to stimulation of the perigenital skin and vaginal tract. Twenty-four mature chinchilla-breed rabbit does were used: 12 to describe the anatomy and innervation of the muscles, 9 to determine reflex EMG activity of the muscles in response to stimulation of the perigenital skin and specific levels of the vaginal tract and 3 to analyze the effect of contraction of the muscles on intravaginal pressure. Both muscles were well developed, with their fibers originating at the ischiadic arch and inserting onto the ligamentum suspensorium clitoridis. Branches of the clitoral and perineal nerves innervated the BSM and ISM, respectively. Bilateral electrical stimulation of these nerves provoked retraction of the clitoral sheath and an increase in intravaginal pressure at the level of the perineal vagina. Whereas neither muscle responded to stimulation of the perigenital skin, both were reflexively activated during mechanical stimulation of the inner walls of the perineal vagina. Prolonged cervical stimulation inhibited this reflex. Thus, in reproductive processes such as copulation and/or parturition, the contraction of these muscles may be induced during stimulation of the perineal vagina. PMID- 11890951 TI - Effects of long-term ingestion of aspartame on hypothalamic neuropeptide Y, plasma leptin and body weight gain and composition. AB - The aim of this study was to determine the effects of the chronic ingestion of aspartame (ASP) on brain neuropeptide Y (NPY) concentrations, plasma hormones, food intake and body fat. Two groups of male Long-Evans rats, fed on a control (C) well-balanced diet, had to drink either a 0.1% ASP solution or water for a period of 14 weeks starting at weaning. Food intake and body weight were weekly recorded. At the end of the experiment, fat pads were sampled, leptin and insulin were measured in the plasma and NPY in several microdissected brain areas. Substituting ASP for water led to lower body weight (-8%; P<.004) and lower fat depot weight (-20%; P<.01) with no differences in energy intake or plasma insulin concentrations. Plasma leptin was significantly reduced by 34% (P<.05). Leptin concentrations were well-correlated with final body weight (r=.47; P<.025) and fat pad mass (r=.53; P<.01). NPY concentrations were 23% lower (P<.03) in the arcuate nucleus of ASP rats with no differences in other brain areas. The beneficial effects on body composition could be related to the decreased effects of NPY on lipid and energy metabolism, independently of insulin. The reasons for the NPY decrease (regulatory or toxicological) are not obvious. The constitutive amino acids of the ASP molecule might participate in the NPY regulation. PMID- 11890952 TI - Dissociation of the effects of nucleus raphe obscurus or rostral ventrolateral medulla lesions on eliminatory and sexual reflexes. AB - Rat preparations were used to investigate long-term changes in external anal sphincter (EAS) contractions and reflexive penile erection following electrolytic lesions of the nucleus raphe obscurus (nRO) or the rostral ventrolateral medulla. EAS contractions were measured electromyographically (EAS EMG) following distention of the EAS with a 5-mm probe. Penile erections were measured using a standard ex copula reflex testing paradigm. At 48 h postlesion, 100% of nRO lesioned animals displayed reflexive erections and the magnitude of EAS EMG was significantly greater in lesioned animals than in sham controls. These results suggested EAS hyperreflexia following destruction of the nRO. By 14 days postlesion, EAS responsiveness in nRO-lesioned animals had returned to levels comparable to nonlesioned animals. No measures of penile erection were affected by nRO lesions. In animals with nucleus gigantocellularis (Gi) and lateral nucleus paragigantocellularis (Gi-lPGi) lesions, no significant changes to EAS reflexes were observed at any time point. At 48 h postoperative, Gi-lPGi lesions significantly reduced the latency to first erection and increased the number of erections elicited relative to controls. Similar facilitation of erection latency was observed at 14 days postlesion, while erection number and flip total were no longer significantly different from controls. These and previous studies suggest that the nRO regulates defecatory reflexes in the rat. These data further suggest that the comingled EAS and bulbospongiosus (BS) motoneurons are controlled by discrete and separate brainstem circuits and that increases in EAS and penile reflexes after spinal cord lesions are mediated by loss of different descending inputs. PMID- 11890953 TI - Experience-induced changes in taste identification of monosodium glutamate. AB - Taste sensitivity for a given subject generally has been thought to be genetically determined and not plastic. Yet experience-inducible changes in human taste and olfactory sensitivities have been reported. To test a taste induction hypothesis, we exposed 17 Americans/Europeans to monosodium glutamate (MSG) in food and then compared their ability to identify MSG taste with that of 2 control groups (18 Americans/Europeans without MSG exposure and 18 Japanese). When tested on Day 11 or 12, the Americans/Europeans exposed to MSG were able to identify MSG at significantly lower concentrations than the Americans/Europeans without MSG exposure. Moreover, Japanese subjects who had prior extensive experience with MSG in Japanese food were able to identify MSG at significantly lower concentrations than the two American/European groups. The differences in identification ability between the two American/European groups challenge the notion of taste sensitivity as stable over time and support the hypothesis of an experience inducible component in human taste. PMID- 11890954 TI - Age-dependent decline in locomotor activity in dogs is environment specific. AB - A decrease in motor activity is an expected concomitant of normal aging and has been reported in humans and nonhuman mammals. We have previously failed to find age differences in open-field locomotor activity in beagle dogs. We now report an age-associated decline when activity measures are taken in the home cage. Locomotor activity of young and aged dogs was examined in both open-field and home-cage environments. Dogs were given six activity tests (two open field, two morning and two afternoon home-cage tests) every second day. Aged dogs were less active than young dogs in the home cage but not in the open field. Activity also varied as a function of sex and housing condition. Behavioral activity is a complex manifestation of many underlying factors. PMID- 11890955 TI - Effect of orosensory stimulation on postprandial thermogenesis in humans. AB - This study assessed the effects of orosensory stimulation by equipalatable stimuli that differed in macronutrient content (lipid and carbohydrate) on postprandial thermogenesis. Sixteen healthy, normal-weight adults (eight males, eight females) participated in six test sessions conducted weekly. The test sessions were administered randomly after overnight fasts and included: ingestion of 50 g of butter in capsules (to avoid oral stimulation with lipids) and 500 ml of water in 15 min followed by no oral stimulation or oral stimulation with a cracker or one of the following foods on a cracker-butter, unsaturated fatty acid (UFA) margarine, jelly, UFA margarine+jelly. Sensory stimulation entailed masticating and expectorating approximately 5.0 g samples of each stimulus every 3 min for 110 min. Blood was drawn immediately after preload ingestion and at minutes 35, 85, 200, 320, and 440 postloading and was analyzed for insulin, glucagon, and glucose. No significant treatment differences were observed for thermogenesis or oxidation of carbohydrate or lipid. Insulin, glucagon, and glucose concentrations were not different between treatments. These data suggest that orosensory stimulation with stimuli differing in lipid and carbohydrate content, but rated similarly in palatability, does not elicit an increased or differential diet-induced thermogenic response. PMID- 11890956 TI - Effects of learned flavour cues on short-term regulation of food intake in a realistic setting. AB - The present study examined the effects of repeated midmorning consumption of novel-flavoured low- and high-energy yoghurt drinks on subsequent energy intake at lunch in 69 adults under actual use conditions. Subjects consumed 200 ml of low- and high-energy yoghurt drinks (67 and 273 kcal/200 ml, respectively), with 20 exposures to each drink on alternate days. Analyses focused on the development of compensation for the differences in energy content of the beverages, due to learned satiety. Results revealed incomplete energy compensation for the beverages, both at first exposure and also after 20 exposures. Relative to the no yoghurt condition, energy intake compensation (+/-S.E.M.) averaged 39+/-36% for the low-energy yoghurt and 17+/-9% for the high-energy version, with no evidence of any change in compensation with repeated exposures. When the flavours of the yoghurt drinks were covertly switched after 20 exposures, subjects increased their energy intake after the high-energy yoghurt drink containing the flavour that was previously coupled with the low-energy yoghurt drink. Vice versa, however, when subjects switched to the low-energy yoghurt drink containing the high-energy flavour, subjects ignored the flavour cue and ate the same lunch size regardless of the energy in the yoghurt drink. We conclude that adults do not readily acquire accurate conditioned adjustments for the energy content in food after repeated experience with the food in free-living natural-eating conditions. PMID- 11890958 TI - Ingestional responses to metabolic challenges in rats selectively bred for high and low saccharin intake. AB - Rats selectively bred on the basis of saccharin intake also differ on some measures of emotional reactivity. The present studies were designed to contribute to our understanding of this association. Rats selectively bred for relatively high (HiS) versus low (LoS) saccharin intake were tested in two paradigms useful in assessing the ability to respond adaptively to internal perturbations of metabolic regulation or to external events that may produce metabolic challenges. The first study concerned slow-onset (regular insulin) and rapid-onset (2-deoxy-D glucose [2-DG], fast-acting insulin) glucoprivation and resultant feeding behavior. LoS and HiS lines did not differ in response to saline or slow-onset challenges, but LoS rats ate less in the first half hour after rapid-onset challenges; the line differences were eliminated by pretreatment with caffeine. The second study revealed significantly higher plasma corticosterone (CORT) among LoS rats relative to HiS rats, both in the light and in the dark. Preliminary assessments after a single stressor and a single dose of dexamethasone showed, respectively, CORT elevation and suppression that was comparable in the two lines. These results add further support to the ideas that voluntary consumption of saccharin is related to the expression of classically defined emotional behaviors, and that responsiveness to diverse metabolic challenges may share a common basis, such as genetic pleiotropism. PMID- 11890957 TI - Interaction of photoperiod and testes development is associated with paternal care in Microtus pennsylvanicus (meadow voles). AB - During the summer breeding season, free-living meadow voles do not engage in paternal care. However, in fall when female territoriality declines, social nesting and breeding activity may overlap and adult males nest with females and young. In the laboratory, meadow voles housed under short day (SD) lengths exhibit more and better quality paternal care than those housed under long day (LD) lengths. This observation is commensurate with the hypothesis that SD paternal care may increase fitness by decreasing pup mortality during colder months. However, SD males also demonstrate variability in paternal care. We hypothesize that this variability may be due to male fertility status; SD infertile males, incapable of siring offspring, should be less likely to care for pups than fertile males, for whom paternal care may confer fitness benefits. The goal of this experiment was to determine whether paternal behavior differed between fertile LD males, fertile SD males (i.e. males that were gonadally photoperiod-unresponsive to SD lengths), and infertile SD males (i.e. males that were gonadally photoperiod-responsive to SD lengths), as indexed by paired testes weights and behavioral evaluation. Fertile SD males exhibited proportionally more paternal behavior than infertile SD males or fertile LD males, which did not differ from each other. Fertile SD males also exhibited paternal behavior faster, spent more time in contact with pups, and engaged in longer and more frequent bouts of pup-directed grooming and huddling than either infertile SD males or fertile LD males. Collectively, these data suggest that photoperiod and fertility status may interact to exert both inhibitory and permissive control over the expression of paternal behavior in adult meadow voles. PMID- 11890959 TI - Physical limitations of the TOBEC method: accuracy and long-term stability. AB - While measurement of electrical conductivity provides one way of estimating body composition in live animals, the accuracy of methods based on this principle requires further study. In this work, we evaluate the effect of the ambient conditions and sample geometry on the response given by an SA-2 model (EM-SCAN) apparatus as well as the reproducibility of the measurements over time. A 2 degrees C variation of the sample temperature (a normal range for living animals) can result in a 6-10% variation in the response. When the conductive mass of a sample is increased in length, the response of the apparatus does not increase once the sample length reaches half the length of the measurement chamber. Samples having the same conductive mass but different shapes can show up to a 17 fold variation in signal. The geometry of the sample itself appears to be of prime importance for determining the strength of the response. We find that the reference phantom provided by the manufacturer is inadequate for calibration and is unable to detect the 10% variation over time of the signal of the apparatus. Until these problems are resolved, the usefulness of the EM-SCAN SA-2 to investigate body composition accurately is questionable. PMID- 11890960 TI - Stressful and behavioral conditions that affect reversible cardiac arrest in the Nile tilapia, Oreochromis niloticus (Teleostei). AB - The objective of the present investigation was to study the reversible cardiac arrest (RCA) to visual stimuli in the unrestrained Nile tilapia (Oreochromis niloticus) as well as the modulation of this response and its behavioral component (arousal/orientation or startle response) by external and internal factors that interfere with alertness and emotionality. The study was preceded by the determination of the autonomic receptors that contribute to the establishment of the heart rate (HR) and the RCA. Systemic injection of atropine and propranolol showed that a double cardiac autonomic control is present in the tilapia. Basal HR was 79.8+/-1.8 beats min(-1) and HR assessed after double autonomic blockade was 74.1+/-3.3 beats min(-1). The mean interbeat interval was 0.79+/-0.40 s during baseline recording and the magnitude of RCA induced by a moving shadow (2.67+/-0.22 s) was higher than that induced by light (1.53+/-1.11 s). RCA is peripherally mediated by muscarinic receptors for it is abolished by atropine but not by propranolol. Stressful conditions like handling the animal outside the water or a nociceptive stimulus (subcutaneous 2% or 3% formalin injection) reduced the cardiac interbeat interval. A subanesthetic dose of barbiturate (5 mg kg(-1)) inhibited RCA induced by a moving shadow stimulus and the startle response, suggesting that an ideal degree of vigilance is necessary for its occurrence. Benzodiazepine injections (1.0 and 2.0 mg kg(-1)) abolished the reduction in magnitude of RCA induced by handling stress and facilitated the startle response, seen in the dry-cold season, in a dose-dependent manner. These data suggest that drugs that act on alertness and on emotionality modulate the magnitude of cardiac interbeat intervals and the corresponding behavioral response. PMID- 11890961 TI - Physiological and behavioral effects of different success during social confrontation in pigs with prior dominance experience. AB - The impact of a 10-h social confrontation on behavior, plasma adrenaline, noradrenaline and cortisol was studied in 14 domestic pigs (eight castrated males and six females) with prior dominance experience. Prior to the experiments, animal groups, each consisting of nine growing pigs (12 weeks old) from different litters, were composed randomly. After 5 days, the pig with the highest rank from each group was removed, provided with a jugular vein catheter and kept in single housing for 2-3 weeks. After this period, each test animal was returned into its familiar group for a 10-h social confrontation. The reintroduction of the test animals into the familiar groups caused frequent agonistic interactions during the first 30 min. Seven animals succeeded to win most of their encounters during the confrontation test and were classified as High Success (HS) animals, whereas seven other animals lost most of their encounters and were classified as Low Success (LS) animals. The reintroduction of the test animals into the groups provoked also marked changes in behavioral and physiological measures. The frequent fighting behavior during the first 30 min was accompanied by a rapid increase of plasma catecholamines and a delayed increase of cortisol. Immediately after introduction, LS pigs tended to show higher plasma adrenaline and noradrenaline concentrations than HS pigs. There was also a tendency for a sustained increase of noradrenaline/adrenaline ratios in HS pigs, whereas the ratios of LS pigs remained nearly unchanged. No significant differences between both groups were found in cortisol levels and in the frequency of agonistic interactions. However, LS animals showed less locomotion, more lying and spent less time exploring the pen or other animals. These results show that during a social confrontation the experience of defeats for formerly high-ranking pigs is accompanied by increased submissive or passive behavior and a higher sympathoneural and adrenomedullary reaction, which may indicate more emotional distress and fear compared to successful animals. PMID- 11890962 TI - Predictable stress promotes place preference and low mesoaccumbens dopamine response. AB - Aversive stimuli that are signaled, and therefore predictable, are preferred to unsignaled ones and promote less severe stress-related disturbances. Since stressful events are known to activate mesoaccumbens dopamine (DA) transmission, in the present experiments, we evaluated possible differences in mesoaccumbens DA response to predictable and unpredictable footshocks. Mice of the inbred strain DBA/2 were trained for conditioned place preference (CPP) in shuttle boxes. The procedure promoted significant preference for the compartment previously paired with predictable shocks (PR) to that paired with unpredictable shocks (NP). Mesoaccumbens levels of DA and its metabolites were therefore evaluated either after the first or the last (third) training session. A significant increase of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels were observed in animals exposed for the first time to the apparatus without shock delivery (SHAM) or to the PR and NP conditions compared with unhandled mice. There was no difference between PR and NP values, and DOPAC and HVA levels in both groups differed from those observable in the SHAM-exposed group. However, trained mice exposed to NP showed a significant elevation of DOPAC and HVA levels in comparison with those exposed to PR. These results show that information about predictability of aversive stimuli reduces central stress responses and suggest a possible relationship between reduced stressfulness and preference for predictable aversive experiences. PMID- 11890963 TI - Diet choice exaggerates food hoarding, intake and pup survival across reproduction. AB - Siberian hamsters increase food intake and hoarding during pregnancy and lactation, perhaps to compensate for large decreases in body fat (approximately 50%). We tested the effects of diet choice on these responses in pregnant, lactating and virgin hamsters housed in a simulated burrow system. Hamsters were offered pellet chow (PC) or a choice of sunflower seeds (SS), rabbit chow (RC) and PC. Pregnant or lactating PC-fed hamsters had increased food intake and hoard size compared with virgins, effects exaggerated by diet self-selection. The pregnancy-induced increases and lactation-induced decreases in body mass were enhanced and diminished by diet self-selection, respectively. Pregnant self selecting hamsters ate relatively more carbohydrate and less fat and hoarded less carbohydrate and more fat than their virgin counterparts (protein not affected). Lactating and virgin self-selecting hamsters both ate and hoarded relatively more carbohydrate than protein or fat compared with PC-fed hamsters but were not different from each other. Litter and pup sizes were similar at birth, but pups from self-selecting mothers had decreased lipid content (50%) compared with pups from PC-fed mothers, whereas at weaning they were heavier but not fatter. Only lactating PC-fed mothers cannibalized their pups (approximately 60% eaten, 8/10 litters). The pregnancy-induced increased eating and hoarding of carbohydrate may have helped meet immediate energy needs sparing dwindling lipid reserves, whereas the decreased fetal lipid investment may have helped conserve energy in anticipation of the increased demands of lactation. The diet-induced exaggerated caloric intake and food hoard size of lactating hamsters may have promoted pup growth and survival. PMID- 11890964 TI - Leptin responsiveness in mice that ectopically express agouti protein. AB - Agouti protein is an endogenous antagonist of melanocortin receptors (MCR), including MCR3 and MCR4, which have been implicated as part of the hypothalamic mechanism that mediates leptin-induced hypophagia. In this experiment we examined the effects of peripheral and central leptin administration in male and female beta-actin promoter (BAPa) mice that express agouti protein ectopically and have a phenotype that includes obesity and diabetes which is exaggerated in males compared with females. Intraperitoneal infusion of 10 microg leptin/day for 13 days caused weight loss and a transient inhibition of food intake in wild-type mice, with a greater effect in males than females. Male BAPa mice were resistant to leptin infusion whereas female mice lost weight. All of the mice lost body weight following a single intracerebroventricular injection of leptin but the effect was greater in female BAPa mice than any other group. There also was a delayed suppression of food intake that was the same for wild-type and BAPa female mice, whereas food intake recovered faster in BAPa than wild-type males. The dissociation between food intake and body weight loss implies a significant effect of leptin on energy expenditure in BAPa mice. These results demonstrate that the effect of leptin on energy balance is not entirely dependent upon the melanocortin system. PMID- 11890965 TI - Rapid noninvasive measurement of hormones in transdermal exudate and saliva. AB - Experiments were conducted in both sheep and humans to evaluate techniques for rapid sampling and measurement of testosterone, insulin, 17-beta estradiol, cortisol and glucose collected in saliva or transdermal exudate. Ultrasound and an electric current facilitated the latter collection. All but insulin were successfully measured in saliva, under resting conditions, and the measured hormones correlated best with blood levels 20-40 min prior to the saliva collection. With imposition of, and recovery from, an exercise stress, this correlation was weakened irrespective of considering the time lag between blood measures during this period and subsequent changes in saliva values. Provided an initial transdermal flux was established, all the hormones and glucose were successfully measured in the transdermal exudate at levels correlating with blood measures at the time of collection, and this held across stressor application and recovery. The transdermal exudate sampling and measurement apparatus is relatively portable, enabling noninvasive collection and analyte measurement, rapidly, at the site where the experiment is being conducted with minimal interference to subjects. This potentially offers a tool of considerable value to endocrine studies. PMID- 11890966 TI - The influence of polysomnography on the Multiple Sleep Latency Test and other measures of daytime sleepiness. AB - INTRODUCTION: According to its guidelines, the Multiple Sleep Latency Test (MSLT) should be performed following an all-night polysomnography (PSG). However, the sleep quality and consequently the MSLT results may be affected by PSG and by the fact that a subject sleeps under unfamiliar conditions. The aim of this study was to examine whether PSG performed in a sleep laboratory has any influence on the MSLT and other measures of daytime sleepiness. METHODS: Twenty healthy subjects with a mean age of 35.9+/-10.1 years underwent two MSLT examinations, and the 2 examination days were at least 4 weeks apart. In addition, on each occasion a monotonous vigilance task (VT) was performed and the subjects were asked to fill out the Epworth Sleepiness (ESS) and Visual Analogue Scales (VAS). In a cross over design, a group of 10 subjects underwent a MSLT (MSLT-P) following a PSG and, on a second occasion, a MSLT (MSLT-N) was performed without a prior PSG. Vice versa, a second group of 10 subjects underwent first MSLT-N and then MSLT-P. RESULTS: None of the MSLT parameters differed significantly between MSLT-P and MSLT-N. The other measures of daytime sleepiness (VT, ESS, VAS) also showed no evidence of significant differences between days with and without a prior PSG. CONCLUSIONS: The results of MSLT and other measures of daytime sleepiness in healthy subjects are not influenced by the fact whether or not the subjects had a PSG the night prior to MSLT. PMID- 11890967 TI - Serum leptin level and restrained eating: study with the Eating Disorder Examination. AB - Serum leptin concentrations in obese patients are influenced at short-term by a reduction of food intake. The objective of this investigation was to evaluate the relationship between serum leptin level and eating behavior. The eating behavior and the food and shape attitudes of a group of obese women were assessed by the Eating Disorder Examination (EDE), and the subscale scores were correlated with serum leptin levels. No difference in serum leptin level was found between obese patients with binge eating disorder (BED) and their nonbingeing counterparts. Considering all patients, the serum leptin levels positively correlated with the body mass index values (BMI), and the EDE subscales scores were positively interrelated. After controlling for BMI, serum leptin level was negatively correlated with the EDE Restraint score and positively correlated with the EDE Shape Concern score. The findings of this investigation indicate that in obese women serum leptin level and the occurrence of binge eating are unrelated. Furthermore, this study also found that the relationship between serum leptin level and restraint over food intake observed in eating disordered patients and in overweight preadolescent girls is shared by obese adult women. In addition, the positive relationship between EDE Shape Concern and serum leptin concentration suggests that the restrained eating might be the cause rather the consequence of the low leptin production. PMID- 11890968 TI - Hepatic vagotomy disrupts somatotropin-induced protein selection. AB - Rats treated with somatotropin (STH) and allowed to self-select between diets varying in protein content will consume more of the high-protein diet. The objective of this study was to determine the role of the hepatic vagus nerve in this ability to select protein. Female Sprague-Dawley rats (n=40) received a hepatic vagotomy (HVAGX) or a sham surgery. Postsurgery, the rats were maintained on pelleted diets for 2 weeks, after which the rats were adapted to selecting between powdered diets with 5% casein and 30% casein. After a 7-day adaptation to diet selection, rats in each surgical treatment group were treated with STH (4 mg/day) or physiological saline for 14 days. Body weight and intake were recorded daily. STH treatment increased growth rate to a similar degree in both sham and HVAGX groups. Despite causing an increase in total food intake, there was no effect of HVAGX alone on body weight. Relative to the sham-saline group, sham-STH in treated rats had greater total food intake that was accounted for entirely by increased consumption of the 30% protein diet and no change in intake of the 5% diet. In contrast, HVAGX+STH rats exhibited 20-30% increases in consumption of both the 5% and 30% protein diets. Thus, the HVAGX+STH rats recognized an increased need for protein, but were unable to distinguish between the high- and low-protein diets and selected more of both. The data suggest that the ability to alter diet selection in response to a stimulation of protein accretion is at least partially mediated through the liver and hepatic branch of the vagus nerve. PMID- 11890969 TI - Iron metabolism in boys involved in intensive physical training. AB - The purpose of the present study was to compare some haematological and iron related parameters of prepubertal boys (10-12 years old) involved in intensive physical training program in preparation for the national championship with nontrained age-matched subjects. For this purpose, iron stores, haemoglobin, ferritin and serum transferrin receptor concentrations were taken into account. The athletes' dietary intakes were similar to the respective intakes in the control group. Mean ferritin concentration was similar in investigated groups of boys at the start of the study, but significant changes between those groups occurred during the 8 months of study. Compared with the control group, the trained ones have shown significantly higher serum transferrin receptor concentration during the competition period of the training season, just when they reached maximum performance capacity. Similarly, significant difference in total body iron, estimated as the sum of the individual's red blood cells iron and iron stores, between investigated groups occurred only during the competition period. The main finding of this study is that the endurance training in boys brings about significant decrease in serum ferritin, as well as iron stores in the body. Haematological parameters and iron status of trained children revealed latent anaemia (15%) or even manifest anaemia (9%). These findings indicate high prevalence of nonanaemic iron deficiency in young athletes and bear relationship to swimming training. PMID- 11890970 TI - Involvement of corticosterone in cardiovascular responses to an open-field novelty stressor in freely moving rats. AB - The aim of the present study was to investigate the modulatory action of different concentrations of circulating corticosterone occupying either predominantly mineralocorticoid receptors (MR) or both MR and glucocorticoid receptors (GR) in control of cardiovascular responses to a novelty stressor. Six groups of rats were instrumented with radiotelemetry transmitters: sham-operated controls, adrenalectomised (ADX) controls, ADX with chronic implantation of a 20 mg corticosterone pellet, ADX with chronic implantation of a 100-mg corticosterone pellet, ADX receiving acute bolus injection of 0.25 mg/kg of corticosterone, and ADX with both implantation of a 20-mg corticosterone pellet and bolus treatment. Exposure to the novelty of an open field caused an increase in blood pressure, heart rate, body temperature and exploratory locomotor activity. The pressor response was dose-dependently increased in ADX rats implanted with a corticosterone pellet. Bolus injection of corticosterone at 10 min prior to novelty had no effect. The tachycardia was reduced in ADX rats compared to sham-operated controls, and this effect was restored by implantation of a 20-mg, but not 100-mg, corticosterone pellet. Bolus injection of corticosterone facilitated the return of heart rate towards baseline levels. The increase in body temperature was reduced in ADX rats, a deficit that was normalised by implantation of either corticosterone dose but not by acute bolus treatment. Locomotor activity was not different between the groups except for a slightly more rapid decline of locomotor activity in both groups treated with a bolus injection of corticosterone. These data show an important role of putative brain MR in maintaining adequate cardiovascular and behavioural responsiveness to a mild psychological stressor, while additional acute or chronic occupation of GR has further differential and sometimes opposing effects. PMID- 11890971 TI - Immediate and long-lasting effects of chronic stress in the prepubertal age on the startle reflex. AB - The immediate and long-lasting effects of two models of chronic stress during the prepubertal period of life (21-32 days) on the acoustic startle response (ASR) were studied in outbred Wistar normotensives and rats with inherited stress induced arterial hypertension (ISIAH) derived from them. Chronic variable stress (CVS) and repeated handling were used as chronic treatment. The obtained data showed a significantly attenuated ASR and a greater magnitude of prepulse inhibition (PPI) in juvenile and adult ISIAH compared to Wistar rats. The immediate effects of prolonged stress on the ASR were genotype-dependent. Young ISIAH rats exposed to both types of prepubertal stimulation had higher ASR than the age-matched controls. No significant stress-induced changes in the ASR were found in young Wistar rats. The long-lasting consequences of prolonged prepubertal stress were similar in the two strains and were determined by the specificity of stress stimulation: chronic handling had no effect on the ASR, while CVS enhanced it. The long-lasting effect of CVS experienced in prepubertal life appears to produce ASR changes similar to those seen in patients with posttraumatic stress disorder (PTSD). The magnitude of PPI increased from early age to adulthood and it was tolerant to environmental influences. The two rat strains did not differ in the rate of short-term habituation to repeated acoustic stimuli, which was unaffected by prepubertal stress. Evidence was obtained indicating that genetic and environmental background in childhood may contribute to the truncation of the startle response. PMID- 11890972 TI - Energy metabolism in women during short exposure to the thermoneutral zone. AB - Ambient temperature has been shown to affect energy metabolism in field situations. Therefore, we assessed the effect of a short exposure to the thermoneutral zone, i.e., 27 degrees C (81 degrees F), in comparison to the usual ambient temperature of 22 degrees C (72 degrees F), on energy expenditure (EE), substrate oxidation, and energy intake (EI) in a controlled situation. Subjects, i.e., women (ages 22+/-2 years, BMI 22+/-3, 28+/-4% body fat), stayed in a respiration chamber three times for 48 h each: once at 22 degrees C, and twice at 27 degrees C in random order, wearing standardized clothing, executing a standardized daily-activities protocol, and being fed in energy balance (EB). During the last 24 h at 22 degrees C, and once during the last 24 h at 27 degrees C, they were fed ad libitum. At 27 degrees C, compared to at 22 degrees C, EE was 8.9+/-1.3 MJ/day vs. 9.9+/-1.5 MJ/day (P<.001) due to decreases in diet-induced thermogenesis (DIT) and activity-induced energy expenditure (AEE) (P<.01); respiratory quotient (RQ) had increased (P<.05); core (P<.05) and skin (P<.001) temperatures had increased. During ad lib feeding, EI was 90-91% of EE (P=.9), due to changes in energy density (ED) of the food choice (P<.01), and related to changes in body temperature and EE (P<.001). Thus, at 27 degrees C, compared to 22 degrees C, energy metabolism was reduced by reductions in DIT and in AEE, while RQ was increased. Reduction in EI was primarily related to body temperature changes and secondarily to changes in EE. PMID- 11890973 TI - Lingual and biting responses to selected lipids by the lizard Podarcis lilfordi. AB - Many lizards can identify food using chemical cues, but very little is known about the chemical constituents used for this purpose. We experimentally investigated responses to several lipid stimuli by the omnivorous lacertid lizard Podarcis lilfordi, which had been shown previously to be capable of identifying prey using only chemical cues and to respond to pork fat by tongue-flicking and biting. In 60-s trials in which stimuli were presented on cotton swabs, the lizards responded very strongly to pure pork fat and to oleic acid, but not to cholesterol or glycerol. Latency to bite swabs, the number of individuals that bit swabs, and the tongue-flick attack score, TFAS(R), which combines effects of tongue-flicks and bites, showed stronger responses to fat than to cholesterol, glycerol, and distilled water but did not differ significantly from responses to oleic acid. Several lines of evidence show that oleic acid elicited strong chemosensory and feeding responses. For individuals that did not bite, the number of tongue-flicks was significantly greater for oleic acid than for distilled water or glycerol, and nearly so for cholesterol. Latency to bite was significantly shorter for oleic acid than for distilled water, and TFAS(R) was significantly greater for oleic acid than for distilled water and glycerol. In combination with pilot data indicating no strong response to the waxy, saturated palmitic acid, these findings suggest that oleic acid in particular and probably other unsaturated fatty acids found in animal fat contribute strongly to the food related responses to lipids. PMID- 11890974 TI - The effects of hypoglycemia and ethanol on rat performance in the radial-arm maze. AB - A deficit in blood glucose (BG) (hypoglycemia, HG) causes a characteristic array of physiological sequelae, symptoms and cognitive impairment in human subjects. However, the performance of hypoglycemic animal subjects in the standard paradigms of behavioral pharmacology has received little research attention. The primary purpose of these experiments was to determine the effect of insulin produced HG on spatial working memory in rats trained in the radial-arm maze (RAM), using a noninterrupted (win-shift) procedure. A second aim was to further investigate possible interaction between HG and ethanol administration since potentiation between the drugs' depressant effects has been reported for rat spontaneous motor activity (SMA). Insulin administration (resulting in BG levels approximately 65% of control levels) was combined factorially with ethanol treatment in two experiments. HG significantly increased time required to complete RAM trials in both experiments, but did not impair accuracy of arm choice. In the second experiment, ethanol was administered only once to minimize development of tolerance, and under these conditions, ethanol at 1500 mg/kg impaired arm-choice accuracy and marginally potentiated HG-produced slowing of running time. The current results appear somewhat similar to previously reported effects of HG on reaction time in human subjects. PMID- 11890975 TI - Heart rate variability: a noninvasive approach to measure stress in calves and cows. AB - The aim of this study was to evaluate the usefulness of heart rate variability (HRV) and its specific parameters as a new approach to assess stress load in cattle. We recorded HRV in 52 calves in three groups and in 31 cows in two groups. In calves we divided Group 1 with no obvious stress load (n=18), Group 2 with external stress load (n=17), and Group 3 with internal stress load from sickness (n=17). In cattle we divided lactating cows (n=21) and nonlactating cows (n=10). HRV parameters were analyzed in the time domain and in the frequency domain. Moreover, we applied Recurrence Quantification Analysis (RQA) to quantify nonlinear components of HRV. In calves, linear HRV parameter decreased from Group 1 to Group 3 (P<.05). However, not a single parameter showed significant differences regarding all three groups. The value of all nonlinear measurements increased at the same time (P<.05). The only parameter that exhibited significant differences between all three groups was the longest diagonal line segment in the recurrence plot (L(MAX)) which is inversely related to the Lyapunov exponent. We did not find differences concerning the linear HRV parameters between the two groups in the cows. The nonlinear parameter Determinism showed significant higher values in lactating cows compared to nonlactating cows. The importance of particular HRV-parameters was tested by applying a discriminant analysis approach. In calves and cattle nonlinear parameters were most important to indicate the level of stress load on the animals. Based on the results we assume HRV to be a valuable physiological indicator for stress load in animals. Whereas linear parameters of HRV are supposed to be useful to separate qualitative different level of stress, nonlinear components of HRV distinguish quantitative different challenges for the animals. PMID- 11890976 TI - Involvement of neuronal nitric oxide synthase in restraint stress-induced fever in rats. AB - Nitric oxide (NO) has been shown to be an important modulator of the febrile response to pyrogens and to psychological stress. In the present study, we aimed to identify the nitric oxide synthase (NOS) isoform (neuronal or inducible, nNOS and iNOS, respectively) involved in restraint stress fever. Colonic temperature (Tc) was measured in unanesthetized rats before and after treatment with the more selective nNOS inhibitor 7-nitroindazole or with the selective iNOS inhibitor aminoguanidine (AG) under unrestrained or restrained conditions. Intraperitoneal injection of AG (25 or 50 mg/kg) did not affect restraint fever, indicating that iNOS is unlikely to be involved in restraint fever. On the other hand, intraperitoneal injection of 7-nitroindazole (25 mg/kg) significantly attenuated the rise in the Tc caused by restraint stress, whereas it caused no change in Tc of euthermic animals. These data show that NO produced by nNOS plays an important role in the genesis of restraint stress-induced fever. PMID- 11890977 TI - Nutrient-secretion coupling in the pancreatic islet beta-cell: recent advances. AB - Insulin secretion from pancreatic islet beta-cells is a tightly regulated process, under the close control of blood glucose concentrations, and several hormones and neurotransmitters. Defects in glucose-triggered insulin secretion are ultimately responsible for the development of type II diabetes, a condition in which the total beta-cell mass is essentially unaltered, but beta-cells become progressively "glucose blind" and unable to meet the enhanced demand for insulin resulting for peripheral insulin resistance. At present, the mechanisms by which glucose (and other nutrients including certain amino acids) trigger insulin secretion in healthy individuals are understood only in part. It is clear, however, that the metabolism of nutrients, and the generation of intracellular signalling molecules including the products of mitochondrial metabolism, probably play a central role. Closure of ATP-sensitive K+(K(ATP)) channels in the plasma membrane, cell depolarisation, and influx of intracellular Ca2+, then prompt the "first phase" on insulin release. However, recent data indicate that glucose also enhances insulin secretion through mechanisms which do not involve a change in K(ATP) channel activity, and seem likely to underlie the second, sustained phase of glucose-stimulated insulin secretion. In this review, I will discuss recent advances in our understanding of each of these signalling processes. PMID- 11890978 TI - The role of synaptic morphology in neural plasticity: structural interactions underlying synaptic power. AB - The study of synaptic plasticity has revealed a common cascade of ultrastructural events across several paradigms. Most notable of these paradigms are development, long-term potentiation (LTP), and adult reactive synaptogenesis (RS). These plastic neural events are discussed in terms of major categories of synaptic morphological change--synaptic density, curvature, and perforations, as well as the size of synaptic elements. The potential functional implications of these morphological changes are reviewed, along with considerations based on recently developed mathematical models of synaptic function. These considerations are then incorporated into the common structural alterations observed during multiple forms of synaptic activation, producing a sequential model supporting increased efficacy associated with neural plasticity. The data suggest that during a plastic challenge, synapses move through a continuum of morphological change, dependent upon the interaction of structural parameters and their effect on various aspects critical to synaptic efficacy. This complex interplay of morphological alterations and synaptic types over time and location may form a critical aspect of neural plasticity. PMID- 11890979 TI - Transcranial magnetic stimulation and Parkinson's disease. AB - While motor cortical areas are the main targets of the integrative activity of basal ganglia, their main output consists of the corticospinal system. Transcranial magnetic stimulation (TMS), a relatively new method to investigate corticospinal physiology, has been widely used to assess possible changes secondary to Parkinson's disease (PD). The use of single- and paired-pulse TMS, two varieties of the original technique, disclosed multiple functional alterations of the corticospinal pathway. For instance, when the latter was tested at 'rest', or in response to somesthetic afferents, it showed excess excitability or reduced inhibition. In turn, during production of a voluntary output, its activation was defective, or inadequately modulated. One major mechanism may be a dysfunction of the interneurons mediating the level of excitation within cortical area 4. For instance, there is a shortening of the so termed 'central silent period', which is a complex, TMS-induced, inhibitory phenomenon possibly mediated by activation of GABA(B) receptors. The so-called 'short-interval intracortical inhibition', which is possibly mediated by GABA(A) receptors, is also diminished. Levodopa restores these and other TMS alterations, thus demonstrating that cortical area 4 is sensitive to dopamine modulation. Overall, TMS has provided substantial new pathophysiological insights, which point to a central role of the primary motor cortex in the movement disorder typical of PD. Repetitive (r-)TMS, another form of TMS, has been studied as a treatment for PD motor signs. Although some reports are favorable, others are not, and have raised the problem of appropriate control experiments. Although extremely interesting, the potential therapeutic role of r-TMS in PD needs further evaluation. PMID- 11890980 TI - Oxidative stress and the prion protein in transmissible spongiform encephalopathies. AB - Transmissible spongiform encephalopathies form a group of fatal neurodegenerative disorders that have the unique property of being infectious, sporadic or genetic in origin. These diseases are believed to be the consequence of the conformational conversion of the prion protein into an abnormal isoform. Their exact pathogenic mechanism remains uncertain, but it is believed that oxidative stress plays a central role. In this article, we will first review in detail the data supporting the latter hypothesis. Subsequently, we will discuss the relationship between the prion protein and the cellular response to oxidative stress, attempting ultimately to link PrP function and neurodegeneration in these disorders. PMID- 11890981 TI - Melanocortins and reproduction. AB - Obesity, anorexia and general body weight fluctuations cause a variety of effects on the reproductive system. Our understanding of the neuro-biological mechanisms of the connections between body weight and the reproductive axis is not especially developed despite a number of interesting physiological observations. Several reports suggest that leptin could play a key role in connecting energy balance and reproduction. The melanocortin system, involving melanocyte stimulating hormone, adrenocorticotrophic hormone, agouti related peptide and the central melanocortin 3 and 4 receptors, plays a major role in the hypothalamic regulation of energy balance. The melanocortins have also been suggested to participate in possible downstream events of the adipose cell derived hormone, leptin. Leptin has importance for several aspects of reproduction including regulation of luteinizing hormone and prolactin release. This review discusses the interplay of hypothalamic regulation of food intake and the hormones involved in the hypothalamic-pituitary-gonadal axis with special emphasis on putative roles of the melanocortin system. PMID- 11890983 TI - Sequelae following traumatic brain injury. The cerebrovascular perspective. AB - Traumatic brain injury (TBI) initiates a huge repertoire of biochemical perturbations. On one hand, destructive events are set into motion while on the other hand, protective and recovery mechanisms are evoked, each with their own temporal and spatial characteristics. The brain exists as a finely tuned balance between vascular, neuronal and glial interactions and so a complex interplay between these factors will dictate the final evolution of pathogenesis. Although vascular damage is a key event, it remains a somewhat neglected component to the underlying degenerative processes that evolve following injury to the brain. The present review will act to integrate the current knowledge of the vascular events proceeding injury to the brain, with an emphasis on how this impacts the control of vascular function and thus cerebral blood flow. PMID- 11890982 TI - The biology of the opioid growth factor receptor (OGFr). AB - Opioid peptides act as growth factors in neural and non-neural cells and tissues, in addition to serving for neurotransmission/neuromodulation in the nervous system. The native opioid growth factor (OGF), [Met(5)]-enkephalin, is a tonic inhibitory peptide that plays a role in cell proliferation and tissue organization during development, cancer, cellular renewal, wound healing, and angiogenesis. OGF action is mediated by a receptor mechanism. Assays with radiolabeled OGF have detected specific and saturable binding, with a one-site model of kinetics. Subcellular fractionation studies show that the receptor for OGF (OGFr) is an integral membrane protein associated with the nucleus. Using antibodies generated to a binding fragment of OGFr, this receptor has been cloned and sequenced in human, rat, and mouse. OGFr is distinguished by containing a series of imperfect repeats. The molecular and protein structure of OGFr have no resemblance to that of classical opioid receptors, and have no significant homologies to known domains or functional motifs with the exception of a bipartite nuclear localization signal. Immunoelectron microscopy and immunocytochemistry investigations, including co-localization studies, have detected OGFr on the outer nuclear envelope where it interfaces with OGF. The peptide-receptor complex associates with karyopherin, translocates through the nuclear pore, and can be observed in the inner nuclear matrix and at the periphery of heterochromatin of the nucleus. Signal transduction for modulation of DNA activity is dependent on the presence of an appropriate confirmation of peptide and receptor. This report reviews the history of OGF-OGFr, examines emerging insights into the mechanisms of action of opioid peptide-receptor interfacing, and discusses the clinical significance of these observations. PMID- 11890984 TI - Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors. Synovial sarcoma. PMID- 11890985 TI - Hereditary breast cancer associated with a germline BRCA2 mutation in identical female twins with similar disease expression. AB - The relative contribution of heritable and nonheritable factors to disease expression in BRCA2 mutation carriers is largely unknown. This report describes a familial breast cancer syndrome in a pair of identical female twins. These twins showed an extremely high concordance in their clinical histories; both twins exhibited similar cancer-related risk factors, and developed breast cancer at the same age with the same disease stage and identical histological features. No differences were detected in hormone receptors status, p53, bcl-2, erbB-2 and LI Ki67 expression by immunohistochemistry. A BRCA2 exon 11 protein truncation test showed a lower molecular weight band than the one expected for a normal allele, in both twins. Sequence analysis of DNA showed a 6 bp insertion between nucleotides 4359-4360, which resulted in a premature stop codon at position 1378. The remarkable disease similarity observed in this identical twin pair is in accordance with an important role for heritable factors in disease expression among patients carrying BRCA germline mutations. PMID- 11890986 TI - A polymorphism in the hMSH2 gene (gIVS12-6T>C) associated with non-Hodgkin lymphomas. AB - DNA common variants may significantly contribute to genetic risk for common diseases. Because of its biological function in DNA repair, hMSH2 gene polymorphisms are candidates for influencing cancer susceptibility and overall genetic stability. Twenty-two individuals with non-Hodgkin lymphomas (NHL) and 50 normal individuals were screened for polymorphic variants in exon 13 of the hMSH2 mismatch repair gene in order to determine if there is any association with development of lymphomas. The polymorphism screening was carried out by single strand conformation polymorphism analysis and DNA sequencing. We found a single nucleotide polymorphism: a T to C substitution at the -6 intronic splice acceptor site of exon 13 (gIVS12-6T>C). This polymorphism was present in 7.5% of normal individuals (allele frequency = 0.05) and in 22.73% of lymphomas (allele frequency = 0.11) (P<0.01). These results suggest that the polymorphism may be associated with an increased risk to develop NHL and that probably there are differences in the effect of the polymorphisms among populations. PMID- 11890987 TI - Replication status in leukocytes of treated and untreated patients with polycythemia vera and essential thrombocytosis. AB - The replication status of malignant cells is usually asynchronous. However, to date the pattern of replication has not been studied in myeloproliferative disorders nor has the effect of chemotherapy been systematically evaluated. Therefore, we used fluorescence in situ hybridization to interphase nuclei in PHA stimulated peripheral blood lymphocytes to examine replication timing of three alleles associated with the malignant process. The study group comprised hydroxyurea treated and untreated patients with essential thrombocytosis (ET) or polycythemia vera (PV). A significantly higher rate of the asynchronous pattern of replication in both treated and untreated patients was found as compared to healthy controls. The highest rate of asynchronous replication was observed in untreated patients. Also, the frequency of the two doublets pattern was significantly higher in the untreated group compared to the treated patients and to the control groups. In conclusion, patients with PV and ET have a higher rate of asynchronous pattern of replication. A possible correlation between disease activity and the pattern of replication is suggested. The effect of hydroxyurea on the pattern of replication is variable. PMID- 11890988 TI - Recurrent chromosome alterations in primary ovarian carcinoma in Chinese women. AB - Ovarian cancer is one of the most frequent gynecological malignancies worldwide with a poor prognosis. Comparative genomic hybridization has been applied to detect recurrent chromosome alterations in 31 primary ovarian carcinomas in Chinese women. Several nonrandom chromosomal changes were identified including gains of 3q (17 cases, 55%) with a minimum region at 3q25 through q26, 8q (16 cases, 52%), 19q (12 cases, 39%), Xq (11 cases, 35%), 1q (10 cases, 32%), 12p12 through q13 (10 cases, 32%), 17q (10 cases, 32%) with a minimum gain region at 17q21, and 20q (9 cases, 29%); and losses of 16q (9 cases, 29%), 1p (7 cases, 23%), 18q (7 cases, 23%), and 22 (7 cases, 23%). High-copy-number amplification was detected in eleven cases. Amplification of 3q25 through q26 was detected in four cases, and amplifications of 8q24 and 12p11.2 through q12 were observed in three cases each. The recurrent gains and losses of chromosomal regions identified in this study provide candidate regions that may contain oncogenes or tumor suppressor genes involved in the development and progression of ovarian cancer. PMID- 11890989 TI - The origin of transformed cells. studies of spontaneous and induced cell transformation in cell cultures from marsupials, a snail, and human amniocytes. AB - Transformation of cells in culture is a model system for carcinogenesis, and two major theories (i.e., mutagenesis and aneuploidy) have emerged from in vitro and in vivo studies. A third view is presented here on the initial steps in the change of primary cells to extended life cells, and their change to immortalized cells. Both changes involve identical, microscopically visible cell abnormalities hitherto dismissed as cell degenerative characteristics. The major cell changes (i.e., giant cells, nuclear fragmentation to form multinucleated cells [MNC]) translated into genetic terms begin with the creation of polyploidy by DNA endoreduplication, followed by amitotic division of these giant cells to produce MNC. Individual nuclei, surrounded by a cell membrane, bud from the surface of the MNC, and represent the origin of the transformed cells. Induced budding by a protease treatment of MNC suggests that the extracellular matrix is an inhibitor of the budding process from human MNC. The production of the MNC is a genetic process determined by two abnormal events (i.e., overproduction of DNA and amitotic chromosomal segregation) during which there are possibilities for different genetic mechanisms to produce inherited variability within and between MNC. These concepts are discussed in regard to carcinogenesis, and by extension its possible prevention by use of the special cytopathic cell changes in carcinogen testing and in clinical screening programs. PMID- 11890990 TI - Cloning of the mRNA of overexpression in colon carcinoma-1: a sequence overexpressed in a subset of colon carcinomas. AB - We have identified a novel human cDNA overexpressed in a colon carcinoma cell line, TC7, established from a tumor with a normal karyotype arising in a patient with a hereditary nonpolyposis colorectal carcinoma. The OCC-1 (overexpressed in colon carcinoma-1) gene is composed of six exons and located in the q24.1 region of chromosome 12. It is transcribed as two mRNA species that differ in their 5'- and 3'-terminal ends. Abundant accumulation of both transcripts was found in placenta, skeletal muscle, kidney, and pancreas tissues. Absent or very faint expression was observed in heart, brain, lung and liver tissues. Overexpressed in colon carcinoma-1 cDNA direct in vitro translation of several polypeptides whose size is shorter than 9 kDa. Attempts to produce antibodies against these synthesized polypeptides in Escherichia coli failed. The absence of sequences at the mRNA and DNA levels hybridizing with mouse sequences together with the absence of a large open reading frame raise the possibility that OCC-1 sequences appeared recently in evolution and are transcribed as two noncoding regulatory RNA. Elevated levels of OCC-1 mRNA were observed in three of eight colon carcinomas as compared to normal mucosa of the same patient. Since these tumors shared the same characteristics of having a near diploid karyotype, OCC-1 overexpression may be a hallmark of this subset of colon carcinomas. PMID- 11890992 TI - Contribution of fluorescence in situ hybridization to immunohistochemistry for the evaluation of HER-2 in breast cancer. AB - The main focus of the present study was to assess the efficacy of interphase cytogenetics using fluorescence in situ hybridization (FISH) as a valid alternative to immunohistochemistry (IHC) in paraffin-embedded tissue sections and/or the efficacy of the combination of the two methods, while, at the same time, aiming to provide additional information on the use of the two methods. For this study, selected breast cancer patients (n=66) were tested for HER-2 gene amplification by FISH. The probe contains DNA sequences specific for the HER-2 human gene locus and hybridizes to the 17q11.2 through q12 region of human chromosome 17. The same samples were tested previously for HER-2 overexpression by two monoclonal antibodies (300G9 and CB11), recognizing an extracellular and an internal domain of gp185(Her-2), respectively. HER-2 overexpression also was evaluated using the HerceptTest Kit (Dako, Milan, Italy). The HerceptTest was performed according to the manufacturer's standard procedures, and results were scored on a 0 to 3+ scale. A total of 34 (51%) of 66 breast tumors enrolled in this study were positive by FISH. Of the 34 cases amplified by FISH, 9 were negative by IHC using both monoclonal antibody (MoAb) 300G9 and MoAb CB11, with a concordance rate from 80.3% to 83.3%. A higher concordance was verified (92.4%) when we used the HerceptTest Kit. Of the 32 cases found negative with the HerceptTest, FISH analysis identified HER-2 gene amplification in more than 10%. Our results indicate that with the combined use of both methods, several amplified samples classified negative by IHC can be used thus improving therapeutic planning for specific therapy with the monoclonal antibody trastuzumab. PMID- 11890991 TI - Novel der(1)t(1;19) in two patients with myeloid neoplasias. AB - Cytogenetic studies can be useful in the clinical management of patients with leukemia. They may also give a clue to leukemogenesis and/or pathogenesis. Numerous disease-specific chromosomal aberrations have been and continue to be identified. Translocation (1;19)(q21 through q23;p13.3) involving the long arm of chromosome 1 and the short arm of chromosome 19 is usually associated with acute lymphoblastic leukemia. We found a new translocation involving one virtually identical breakpoint 19p13 and one distinct 1p13 in two cases of myeloid neoplasms. Studies of bone marrow and peripheral blood specimens specified in one of our patients acute myeloid leukemia and in an other myelodysplastic syndrome. Conventional cytogenetics was supplemented by spectral karyotyping (SKY), microdissection, and fluorescence in situ hybridization. Our first case showed a der(1)t(1;19)(p13;p13.1) as the sole chromosomal change. In addition to this translocation, a pericentric inversion within chromosome 10 and with a cryptic t(10;11) were detected by SKY in the second case. Translocation (1;19)(p13;p13.1) may play a role in the leukemogenesis of myeloid diseases. PMID- 11890993 TI - EWS/FLI-1 fusion signal inserted into chromosome 11 in one patient with morphologic features of Ewing sarcoma, but lacking t(11;22). AB - A reciprocal t(11;22)(q24;q12) is found in 85% of Ewing sarcomas (ES) cases. We report a case of a child with ES, in whom trisomy 8 was observed as the sole chromosomal abnormality. Fluorescence in situ hybridization---using a set of probes that localize to 22q12 (EWS) and 11q24 (FLI-1) and usually show the translocation as fusion (red-green) signal on der(22)---showed a fusion signal on der(11) suggesting an insertion as the mechanism that led to the EWS-FLI-1 gene rearrangement. Reverse transcriptase-polymerase chain reaction studies revealed the presence of two EWS/FLI1 fusion gene products. PMID- 11890994 TI - Defining regions of loss of heterozygosity of 16q in breast cancer cell lines. AB - The loss of heterozygosity (LOH) of chromosome 16 was assessed in 21 breast cancer cell lines and two nontumorigenic breast epithelial cell lines by typing microsatellite markers distributed on this chromosome. In addition, dual-color fluorescence in situ hybridization was used to metaphase spreads of these cell lines using chromosome 16 paint and region specific probes. Eleven of the cell lines had LOH for chromosome 16, two for the entire chromosome, three for the long arm, and six had LOH for restricted regions of the long arm. The results supported evidence that there are two predominant regions of LOH, 16q22.1 and 16q24.3. The cell lines with chromosome 16 LOH can be used for screening candidate tumor suppressor genes at 16q in breast cancer. PMID- 11890995 TI - Identification of ins(8;21) with AML1/ETO fusion in acute myelogenous leukemia M2 by molecular cytogenetics. AB - A high percentage of cases of acute myelogenous leukemia (AML) of the M2 subtype show a rearrangement between the AML1 and ETO genes. The detection of the AML1/ETO fusion has clinical relevance because patients with this subtype have a good prognosis. We present the results of conventional and molecular cytogenetic studies in a patient with acute myelogenous leukemia French-American-British M2 classification, who had a complex karyotype involving chromosomes 8 and 21. Dual color fluorescence in situ hybridization (FISH) using the AML1/ETO probe demonstrated a recombination of both genes on an add(8) chromosome. The use of other FISH probes (CEP8, c-myc and TEL21) and spectral karyotyping indicated that AML1/ETO fusion occurred as a consequence of a previously undescribed ins(8;21)(q22;q22.1q22.3). PMID- 11890996 TI - A novel dic(1;10) in a patient with myelodysplastic syndrome. AB - We report on a case of refractory anemia with trilineage dysplasia and an unbalanced der(1)t(1;10) that resulted in trisomy of the long arm of chromosome 1 (1q) and monosomy of the short arm of chromosome 10 (10p). Fluorescence in situ hybridization showed that the rearranged chromosome contained the centromeres of both chromosomes 1 and 10, leading to a dic(1;10). To our knowledge, a dicentric chromosome involving chromosomes 1 and 10 has never been described in hematological malignancies. PMID- 11890997 TI - Absence of HMGIC-LHFP fusion in pulmonary chondroid hamartomas with aberrations involving chromosomal regions 12q13 through 15 and 13q12 through q14. AB - In a variety of benign solid human tumors the high mobility group protein gene HMGIC is affected by aberrations involving the chromosomal region 12q14 through q15. Beside the two predominant alterations t(3;12) (q27 through 28;q14 through q15) and t(12;14)(q14 through q15;q23 through q24), the t(12;13)(q14 through q15;q12 through q14) is another aberration observed recurrently in these tumors. Very recently, an HMGIC-LHFP (lipoma HMGIC fusion partner) fusion gene has been detected in a lipoma with a t(12;13). The results of the present study demonstrated the absence of the HMGIC-LHFP fusion in three pulmonary chondroid hamartomas (PCH) with complex aberrations involving chromosomal regions 12q13 through q15 and 13q12q through q14 and one PCH with a simple t(12;13)(q14 through 15;q13) by reverse transcription-polymerase chain reaction. Thus, intragenic rearrangements within the LHFP gene leading to its fusion to HMGIC are not a consistent finding in mesenchymal tumors with clonal aberrations of both chromosomal regions 12q13 through q15 and 13q12 through q14. PMID- 11890998 TI - Interphase fluorescence in situ hybridization and DNA flow cytometry analysis of medulloblastomas with a normal karyotype. AB - Interphase fluorescence in situ hybridization (FISH) with chromosome 3 and 17 centromeric probes and DNA flow cytometry were used for a retrospective study of nine pediatric medulloblastomas with normal karyotypes after tissue culture. The FISH analysis of medulloblastoma touch preparations showed that in seven of nine tumors, a significant proportion of nuclei had an increased number of centromeric signals for the selected chromosomes. In six of seven cases, this increase was caused by the presence of triploid and tetraploid clones as established by flow cytometry of paraffin-embedded tumors. These findings show that molecular cytogenetic analysis combined with DNA flow cytometry is necessary for all pediatric medulloblastomas diagnosed as cytogenetically normal on cultured tumor tissue. PMID- 11890999 TI - Detection of minimal residual disease in peripheral blood stem cells from two acute myeloid leukemia patients with trisomy 8 predicts early relapse after autologous bone marrow transplantation. AB - We report two cases of acute myeloid leukemia (AML) French-American-British M4 classification with trisomy 8 at diagnosis as the sole chromosome abnormality. Both patients were treated with the GIMEMA AML-10 protocol and underwent autologous bone marrow transplantation (ABMT) in hematologic remission. Peripheral blood stem cells (PBSC), and bone marrow in one patient, were collected after consolidation therapy and tested by fluorescence in situ hybridization (FISH) analysis with an alpha-satellite probe for chromosome 8. It revealed that all samples were positive for minimal residual disease (MRD) as the value of trisomic cells exceeded the mean +3 standard deviations of the controls. ABMT was done following a myeloablative regimen (busulphan/cyclophosphamide) and PBSC were reinfused. Both patients relapsed, 4 and 2 months, respectively, after autotransplant. Although more data are needed, these results suggest that the persistence of MRD, as detected by FISH, in stem cell collections, is associated with a poor outcome in AML patients with trisomy 8 undergoing ABMT. PMID- 11891000 TI - Cytogenetic characterization of Ewing tumors: further update on 20 cases. PMID- 11891001 TI - Effects of nonylphenol on the gonadal differentiation of the hermaphroditic fish, Rivulus marmoratus. AB - Nonylphenol (NP) is an estrogenic degradation product of alkylphenol polyethoxylate surfactants. In this study, the effects of NP on gonadal differentiation and development in Rivulus marmoratus (Osteichthyes, Cyprinodontiformes), a self-fertilizing, hermaphroditic species, were examined. Starting at hatching, fish were exposed to 150 or 300 microg 1(-1) NP (nominal concentrations) in a static system with daily renewal. The measured concentration of NP in the test water decreased rapidly; half-life was 8.0 h. After 60 d of exposure to NP, fish were kept in uncontaminated water for 20 d and were then preserved for histological examination. No fish exposed to 300 microg l(-1) NP (N=8) and only two of nine fish exposed to 150 microg l(-1) NP developed testicular tissue, compared with nine of 13 water-control fish and five of nine solvent-control fish. Oogenesis was also significantly inhibited by NP. None of the fish exposed to 300 microg l(-1) and only two of nine fish exposed to 150 microg l(-1) NP had vitellogenic oocytes, compared with seven of 11 water-control fish (not including males) and six of nine solvent-control fish. Dysplasia of the gonadal lumen also occurred in fish exposed to 300 microg l(-1) NP. These changes, including testicular agenesis, have not been previously reported in fish exposed to NP. PMID- 11891002 TI - Effects of cadmium on nuclear integrity and DNA repair efficiency in the gill cells of Mytilus edulis L. AB - Although the effects of heavy metals on marine invertebrate species are well studied in term of their toxicity and bioaccumulation, less is known about their genotoxicity. The aim of this investigation was to assess the DNA damaging potential of cadmium (Cd) in an important pollution sentinel organism, the mussel Mytilus edulis. Cadmium is one of the most toxic and widespread heavy metals found in the marine environment, and is a recognised carcinogen in mammals. Based on the results of the comet assay (alkaline single cell gel electrophoresis), Cd was found not to be genotoxic in mussel gill cells under acute and chronic exposure conditions, whereas pre-exposure to low concentrations of Cd was found to enhance the genotoxicity of another mutagen, hydrogen peroxide (H2O2). The effects of H2O2 were normally reversible when cells were transferred to clean saline buffer. However, in cells that had been pre-treated with Cd, in vivo or in vitro, we observed a decrease in this post-treatment DNA repair. The effects of Cd were reversed by zinc which suggests that the inhibitory effect of Cd on DNA repair was due to the displacement of zinc ions from active sites on proteins involved in the repair process (a property already described for mammals). Moreover, since Cd inhibits or delays the onset of apoptosis (programmed cell death), this removes one of the main defence mechanisms responsible for protecting the organism against neoplasia. There appears to be a close similarity between the effects of Cd on marine molluscs and mammals. PMID- 11891003 TI - Sublethal actions of copper in abalone (Haliotis rufescens) as characterized by in vivo 31P NMR. AB - The sublethal biochemical actions of copper in live, intact red abalone (Haliotis rufescens) were characterized by in vivo 31P nuclear magnetic resonance spectroscopy (NMR). This non-invasive technique is ideal for examining cellular respiration since critical metabolite concentrations, including phosphoarginine ([PA]), inorganic phosphate ([P(i)]) and [ATP], and the arginine kinase (AK) rate constant, can be monitored in real time. Both metabolite concentrations and enzyme rate constants were measured in abalone during 8-h exposures to 66 microg l(-1) (1.04 microM) and 126 microg l(-1) (1.98 microM) copper (as CuCl2). Significant decreases in [PA] and corresponding increases in [P(i)] resulted, while [ATP] remained constant. In controls [PA], [P(i)] and [ATP] all remained unchanged. Furthermore, both copper concentrations induced a significant elevation in the forward AK rate constant over the basal value of 0.020 +/- 0.002 s(-1). Metabolite levels and enzyme rate constants were also measured during 8-h 66 microg l(-1) copper exposures both before and after a 2-week subchronic exposure to 36 microg l(-1) (0.57 microM) copper. Unlike before the subchronic exposure, no significant changes in [PA], [P(i)] or [ATP] were observed after the 36 microg l(-1) copper treatment, compared with controls. This induced tolerance was also evident from the forward AK rate constant data. Finally, copper accumulation was determined in gill, digestive gland and foot muscle samples. Whereas acute exposure only led to significant accumulation in the gill, copper levels in subchronically exposed abalone were significantly elevated in both the gill and digestive gland, and marginally so in foot muscle. Overall, the gill appears to be the primary site of copper accumulation and toxicity, while the foot and adductor muscles maybe secondarily impacted. The observed metabolic changes may result from insufficient oxygen delivery to the muscles, resulting from mucus accumulation or cytological damage at the gill. In conclusion, abalone acutely exposed to copper pollution may develop asphyxial hypoxia. Since their survival is dependent on adherence to rock surfaces, such a reduction of muscle function could ultimately prove fatal. PMID- 11891004 TI - Accumulation and distribution of dietary arsenic in lake whitefish (Coregonus clupeaformis). AB - Adult lake whitefish (Coregonus clupeaformis) were fed As contaminated diets at nominal concentrations of 0, 1, 10 and 100 microg As/g food (d.w.) for 10, 30 and 64 days. Liver, kidney, stomach, intestine, pyloric caeca, gallbladder, skin and scales were analyzed for As content. The pattern of As accumulation in fish tissues was influenced by reduced feed consumption beginning on day 45 by fish fed the 100 microg As/g food. Significant As accumulation occurred in all tissues examined from fish exposed to the 100 microg As/g food for 30 days, with the exception of gallbladder. After 30 days of exposure, the highest concentration of As was observed in pyloric caeca of fish fed the 100 microg As/g food. Significant accumulation of As occurred in livers and scales of fish fed concentrations of As as low as 10 microg/g for 30 and 64 days. Muscle, gonad, spleen, gills and bone of lake whitefish fed a control diet for 10 days and 100 microg As/g food for 10, 30 and 64 days were also analyzed for As content. As concentrations increased in gonads, spleen and gills of fish fed the 100 microg As/g food for 30 days. Increased concentrations of As were observed in bone of fish fed the high dose food after each duration of exposure. As concentrations did not increase in muscle of fish after 10, 30 or 64 days of exposure. The following manuscript (Pedlar et al., 2001) documents toxicological effects observed in these fish. Analyses of As in pyloric caeca, intestine, liver and scales are recommended to evaluate the bioavailability of As to freshwater fish in environmental monitoring programs. PMID- 11891005 TI - Toxicological effects of dietary arsenic exposure in lake whitefish (Coregonus clupeaformis). AB - Adult lake whitefish were fed As contaminated diets at nominal concentrations of 0, 1, 10, and 100 microg As/g food (dry weight) for 10, 30, and 64 days. Reduced feed consumption was observed in lake whitefish fed the 100 microg As/g food, beginning on day 45 of exposure. The accumulation and distribution of As in these fish are described in the previous manuscript [Pedlar, R.M., Klaverkamp, J.F., 2001. The accumulation and distribution of dietary arsenic in lake whitefish (Coregonus clupeaformis). Aquat. Toxicol., in press]. At the molecular level of organization, metallothionein (MT) induction occurred in lake whitefish fed the 100 microg As/g food after 10 and 30 days, and in fish fed the 1 and 10 microg As/g diets for 64 days. Dietary As exposure did not have a significant effect on plasma lipid peroxide (LPO) concentrations. At the tissue and organ level, mean liver somatic index decreased significantly in lake whitefish fed the 100 microg As/g food for 64 days. Blood parameters (hematocrit, hemoglobin concentration, red blood cell count, mean cell volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration) were not affected by exposure to As contaminated diets. Liver and gallbladder histopathologies were observed in lake whitefish fed all As contaminated diets after each duration of exposure. Histopathology observed in liver included nuclear, architectural and structural alterations, areas of inflammation, and focal necrosis. Sloughing of the epithelium, dilation of vascular elements, inflammation, edema, fibrosis, and increased width of the submucosa were some of the alterations observed in gallbladders of lake whitefish fed As contaminated diets. Both organs were sensitive to As exposure, as damage occurred with exposure to dietary concentrations of As as low as 1 microg/g. Whole organism parameters were unaltered by dietary As exposure. Based on the results of this study, histopathological alterations in liver and gallbladder, and hepatic MT induction may be useful indicators of As toxicity in environmental monitoring programs that also measure As concentrations in those tissues. PMID- 11891006 TI - Effects of food ration on survival and sublethal responses of lake chubsuckers (Erimyzon sucetta) exposed to coal combustion wastes. AB - Study organisms in chronic toxicological bioassays are often provided with excessive resources to remove food limitations as a confounding experimental variable. Under more ecologically realistic situations, resources are often less abundant and such restrictions may alter the responses of organisms to environmental contaminants. Here, we investigated the interaction between resource level and sediment toxicity in the lake chubsucker, Erimyzon sucetta. For 78 days we fed fish one of three ration levels (1X, 2X, 4X; uncontaminated food) that was grazed directly from either clean sand or coal ash-contaminated sediments. Despite provision of uncontaminated food, fish exposed to the contaminated sediments accumulated significant whole body concentrations of As, Se, Sr, and V. Food ration affected the pattern of Se accumulation, with lowest concentrations accumulated by fish supplied with the lowest rations (1X). Paradoxically, fish in the 1X-ash treatment were most adversely effected by ash exposure, despite having Se burdens much lower than fish in the 2X- and 4X-ash treatments. Fish in the 1X-ash treatment exhibited higher mortality, lower proportional growth, and increased incidence of fin erosion compared to fish provided with higher rations. Such results may, in part, be explained by the apparent inability of fish with reduced rations to maintain positive energy balance, as evidenced by their higher standard metabolic rates compared to control fish fed similar rations. Our results underscore the importance of considering resource quantity and nutritional factors in chronic bioassays in order to draw more ecologically realistic conclusions about contaminant effects. PMID- 11891007 TI - Ghrelin--not just another stomach hormone. AB - Growth hormone (GH) secretagogues (GHSs) are non-natural, synthetic substances that stimulate GH secretion via a G-protein-coupled receptor called the GHS receptor (GHS-R). The natural ligand for the GHS-R has been identified recently; it is called ghrelin. Ghrelin and its receptor show a widespread distribution in the body; the greatest expression of ghrelin is in stomach endocrine cells. Administration of exogenous ghrelin has been shown to stimulate pituitary GH secretion, appetite, body growth and fat deposition. Ghrelin was probably designed to be a major anabolic hormone. Ghrelin also exerts several other activities in the stomach. The findings that ghrelin is produced in mucosal endocrine cells of the stomach and intestine, and that ghrelin is measurable in the general circulation indicate its hormonal nature. A maximal expression of ghrelin in the stomach suggests that there is a gastrointestinal hypothalamic pituitary axis that influences GH secretion, body growth and appetite that is responsive to nutritional and caloric intakes. PMID- 11891008 TI - 125I[Sar(1)Ile(8)] angiotensin II has a different affinity for AT(1) and AT(2) receptor subtypes in ovine tissues. AB - Iodinated angiotensin II (Ang II) and its analogues are often assumed to have equal affinities for AT(1) and AT(2) receptor subtypes. However, using saturation and competition binding assays in several tissues from pregnant, nonpregnant, and fetal sheep, we found the affinity of 125I[Sar(1)Ile(8)] Ang II for Ang II receptors was different (P<0.05) between tissue types. The dissociation constants (Kd) and half maximal displacements of [Sar(1)Ile(8)] Ang II (Sar IC(50)) were directly related (P<0.05) to proportions of AT(1) receptors, and inversely related (P<0.05) to proportions of AT(2) receptors in tissues from all groups combined, in tissues from individual groups (pregnant, nonpregnant or fetal), and in some individual tissues (uterine arteries and aortae). This suggests that 125I[Sar(1)Ile(8)] Ang II has a different affinity for AT(1) and AT(2) receptors in ovine tissues. The Kds of 125I[Sar(1)Ile(8)] Ang II for "pure" populations of AT(1) and AT(2) receptors were 1.2 and 0.3 nM, respectively, i.e. affinity was four-fold higher for AT(2) receptors. We corrected the measured proportions of the receptor subtypes using their fractional occupancies. In tissues which contained at least 10% of each receptor subtype, the corrected proportions were significantly altered (P<0.05), even in some tissues, to the extent of being reversed. PMID- 11891010 TI - Formation of angiotensin-(1-7) from angiotensin II by the venom of Conus geographus. AB - The binding of [3H]angiotensin II to AT(1) receptors on Chinese Hamster Ovary cells expressing the human AT(1) receptor (CHO-AT(1) cells) is potently inhibited by venoms of the marine snails Conus geographus and C. betulinus. On the other hand, the binding of the nonpeptide AT(1) receptor-selective antagonist [3H]candesartan is not affected but competition binding curves of angiotensin II and the peptide antagonist [Sar(1),Ile(8)]angiotensin II (sarile) are shifted to the right. These effects resulted from the breakdown of angiotensin II into smaller fragments that do not bind to the AT(1) receptor. In this context, angiotensin-(1-7) is the most prominent fragment and angiotensin-(1-4) and angiotensin-(1-5) are also formed but to a lesser extent. The molecular weight of the involved peptidases exceeds 50 kDa, as determined by gel chromatography and ultrafitration. PMID- 11891009 TI - N-terminal chromogranin-derived peptides as dilators of bovine coronary resistance arteries. AB - N-terminal peptides of chromogranin A and B (CGA and CGB) were compared for dilator responses in isolated bovine coronary arteries (bCoA), measuring diameter changes as a function of pressure. bCoA developed and maintained myogenic tone (MYT) at approximately 20% from 50 to 150 mm Hg. In contrast to CGB(1-40), CGA(1 40) and CGA(1-76) (VS-I) both displayed significant intrinsic vasodilator effects. CGA(1-40) reduced myogenic reactivity from 70 to 150 mm Hg (p<0.05, n=6). At 75 mm Hg, CGA(1-40) showed a concentration-dependent dilatation at 0.1 nM-10 microM. The dilator effect of CGA(1-40) persisted at moderately elevated [K(+)](e) (8.4-16 mM). However, this effect was diminished by pertussis toxin (PTX) and abolished by antagonists to several subtypes of K(+) channels (tetraethylammonium, Ba(2+) and glibenclamide). These results demonstrate that the N-terminal domain of CGA has dilator effect in the myogenically active bCoA. We propose that CGA(1-40) and the naturally occurring vasostatin I are regulatory peptides of relevance for the coronary microcirculation and that a G(alphai) sub unit and K(+) channel activation may be involved in the signal pathway. PMID- 11891012 TI - Down-regulation of corticotropin-releasing hormone receptor type 2beta mRNA expression in the rat cardiovascular system following food deprivation. AB - The present study was conducted to assess the effect of nutritional stress induced by food deprivation on expression of messenger ribonucleic acid (mRNA) for corticotropin-releasing hormone receptor type 2beta (CRH-R2beta) in the rat cardiovascular system in the presence or absence of changes in circulating corticosterone. Food deprivation for 96 h caused a robust increase in plasma corticosterone levels and a significant decrease in CRH-R2beta mRNA expression in the rat heart. Starvation for 48 and 96 h decreased CRH-R2beta mRNA expression in the atria, ventricle as well as aorta of sham-adrenalectomized (sham) rats. Surprisingly, clamping plasma glucocorticoids at low levels by adrenalectomy with corticosterone pellet replacement (ADX+B) did not completely prevent starvation induced decreases of CRH-R2beta mRNA expression in the rat cardiovascular system. Urocortin (Ucn) mRNA expression was increased significantly by food deprivation in the heart of sham as well as ADX+B rats. We speculate that food deprivation may increase urocortin, which in turn down-regulates CRH-R2beta mRNA expression in cardiovascular system. These data indicate that food deprivation despite the presence or absence of changes in circulating corticosterone may have an inhibitory effect on CRH-R2beta mRNA expression in the rat cardiovascular system. PMID- 11891011 TI - Parathyroid hormone-related protein enhances PC-3 prostate cancer cell growth via both autocrine/paracrine and intracrine pathways. AB - Parathyroid hormone-related protein (PTHrP) is expressed by human prostatic tissue and prostate cancer cell lines, and positively influences primary prostate tumor growth in vivo. The human prostate cancer cell line PC-3, which expresses functional PTH/PTHrP receptors, was used as a model to study the effects of PTHrP on prostate cancer cell growth. Addition of PTHrP (1-34), (1-86), and (1-139) increased cell number and [3H]thymidine incorporation; these effects were reversed by anti-PTHrP antiserum. This antiserum also decreased endogenous PC-3 cell growth. Clonal PTHrP-overexpressing PC-3 cell lines also showed enhanced cell growth and [3H]thymidine incorporation and were enriched in the G2+M phase of the cell cycle, suggesting an effect of PTHrP on mitosis. Overexpression of PTHrP with the nuclear localization sequence (NLS) deletion partially reversed the growth-stimulatory effects. The growth rate of these cells was midway between that of wild-type PTHrP-overexpressing and control cells, presumably because NLS mutated PTHrP is still secreted and acts through the cell surface PTH/PTHrP receptor. In contrast to NLS-mutated PTHrP, wild-type protein showed preferential nuclear localization. These results suggest that the proliferative effects of PTHrP in PC-3 cells are mediated via both autocrine/paracrine and intracrine pathways, and that controlling PTHrP production in prostate cancer may be therapeutically beneficial. PMID- 11891014 TI - Atrial natriuretic factor as marker of myocardial compromise in Chagas' disease. AB - This study investigated the evolution of plasma atrial natriuretic factor (ANF) in patients in different stages of Chagas' disease and analyzed its usefulness as prognostic factor of the development of myocardial compromise in asymptomatic chagasic patients. Chagas' disease, a determinant of heart failure, is caused by the parasite Trypanosoma cruzi. A total of 21 chagasic patients were studied: 9 in the asymptomatic stage, 6 with conduction defects (CD), and 6 with chronic heart failure (CHF); and 31 controls: 16 healthy, 6 with CD, and 9 with CHF. Plasma ANF radioimmunoassay (RIA) and complementary studies were performed twice for each patient, with an interval period of 12 months. First sample: chagasic patients showed higher ANF levels in the CHF group than in CD and asymptomatic subjects; second sample: the peptide levels were higher in CHF patients than in the asymptomatic group. In non-chagasic CHF patients, ANF levels were higher than in CD patients and controls in both samples. ANF levels were not able to differentiate chagasic asymptomatic and CD patients from healthy subjects and CD controls; meanwhile, chagasic CHF patients showed lower plasma ANF than their controls. Furthermore, ANF is a sensitive marker capable of detecting gradual impairments in cardiac function in all patients studied. PMID- 11891013 TI - Low-affinity CCK-1 receptors inhibit bombesin-stimulated secretion in rat pancreatic acini--implication of the actin cytoskeleton. AB - EXPERIMENTAL OBJECTIVES: Stimulation of low-affinity CCK-1 receptors on pancreatic acini leads to inhibition of enzyme secretion. We studied signal transduction mechanisms to identify potential causes for the reduced secretion. RESULTS: Co-stimulation experiments with CCK, CCK-JMV-180, and bombesin revealed an inhibition of bombesin-stimulated enzyme secretion by low-affinity CCK-1 receptors. Binding of 125I-gastrin-releasing peptide (the mammalian analogue of bombesin) to acini after CCK preincubation was not altered. After a short preincubation of acini with high concentrations of CCK, intracellular calcium remained responsive to bombesin. In contrast to bombesin or CCK at concentrations of 10(-10) M or lower, high concentrations of CCK caused a strong activation of p125 focal adhesion kinase (p125(FAK)) and a marked reorganisation of the actin cytoskeleton. CONCLUSIONS: Inhibitory mechanisms triggered by low-affinity CCK-1 receptors interrupt enzyme secretion from pancreatic acini at late stages in the signal transduction cascades since bombesin receptor binding and early signalling events remained intact after CCK preincubation. A reorganisation of the actin cytoskeleton is suggested to be the mechanism by which low-affinity CCK-1 receptors actively interrupt enzyme secretion stimulated by other receptors. PMID- 11891015 TI - Troglitazone ameliorates lipotoxicity in the beta cell line INS-1 expressing PPAR gamma. AB - To elucidate the mechanisms by which troglitazone, which is a direct ligand for peroxisome proliferator-activated receptor (PPAR) gamma, ameliorates insulin resistance, we have demonstrated that PPAR gamma is expressed in a pancreatic beta cell line, INS-1, using reverse transcription-polymerase chain reaction (RT PCR). We incubated the cells with 5 micromol/l troglitazone and 1 mmol/l of each major free fatty acid (FFA; palmitic acid, oleic acid, and linoleic acid), alone or in combination, for 48 h. After that, we evaluated glucose-stimulated insulin secretion (GSIS) and 25 mmol/l KCl-induced insulin secretion in the presence of diazoxide, which clamps membrane potential. Our results showed: (1) treatment with troglitazone for 48 h caused enhancement of GSIS, although troglitazone significantly suppressed cell viability assessed by MTT assay. (2) In cells co treated with troglitazone and FFA, troglitazone ameliorated lipotoxicity due to FFA. (3) In the presence of 300 micromol/l diazoxide and 25 mmol/l KCl, troglitazone did not affect the recovery of GSIS in INS-1 cells. These results suggest that insulin secretion from the rat insulinoma cell line, INS-1, is modulated by troglitazone, acting somewhere in the ATP-sensitive K(+) channel pathway, possibly through PPAR gamma. PMID- 11891016 TI - Tumor necrosis factor system in insulin resistance in gestational diabetes. AB - OBJECTIVE: The aim of the study was to investigate the pathophysiological role of the tumor necrosis factor (TNF) system in insulin resistance in patients with gestational diabetes (GDM) and during the course of normal pregnancy. PATIENTS AND METHODS: Thirty women with GDM (16-39 gestational weeks), 35 healthy pregnant women (15 first, nine second and 11 third trimester) and 25 healthy age-matched non-pregnant women were studied. Serum TNF-alpha, and its soluble receptors 1 and 2 (sTNFR-1 and -2) were measured. RESULTS: In non-diabetic pregnant women in the third trimester all measures were significantly higher (P<0.05 or less) than in the first trimester and in non-pregnant women (BMI 27.6 +/- 4.1 (+/- S.D.), 24.1 +/- 2.6, 22.4 +/- 2.4 kg/m(2)), serum TNF-alpha (4.6 +/- 0.6, 4.1 +/- 0.4, 4.1 +/ 0.4 ng/l), sTNFR-1 (2.7 +/- 0.9, 2.0 +/- 0.5, 2.0 +/- 0.1 microg/l), sTNFR-2 (5.6 +/- 2.6, 4.6 +/- 2.1, 3.3 +/- 0.2 microg/l), C-peptide (3.1 +/- 1.7, 1.1 +/- 0.7, 1.1 +/- 0.8 microg/l), and C-peptide:blood glucose ratio (0.6 +/- 0.2, 0.2 +/- 0.1, 0.2 +/- 0.1 microg/mmol). In GDM these measures were even higher than in any subgroup of healthy pregnant women (BMI) (33.4 +/- 6.4 kg/m(2), TNF-alpha) (6.3 +/- 0.6 microg/l), sTNFR-1 (3.0 +/- 0.5 microg/l), sTNFR-2 (10.0 +/- 6.9 microg/l, C-peptide 6.0 +/- 2.7 microg/l, C-peptide:blood glucose ratio: 1.2 +/- 0.5 microg/mmol, P<0.01). Significant (P<0.01) positive linear correlations were found in gestational diabetic and non-diabetic women between serum TNF-alpha, C peptide levels, and BMI. In gestational diabetic women, in multivariate analysis studying the dependency of C-peptide only BMI remained significant (r(2)=0.67, P=0.01). CONCLUSIONS: Our observation emphasizes the obesity-related component of insulin resistance driven by adipocytokines, such as TNF-alpha and its receptors during the course of normal pregnancy and GDM. PMID- 11891017 TI - Glucagon-like peptide-1 response to acarbose in elderly type 2 diabetic subjects. AB - The anti-hyperglycemic effect of alpha-glucosidase inhibitors (AGI) is partly attributed to their ability to stimulate the secretion of glucagon-like peptide-1 (GLP-1), a gut hormone with insulin stimulating capability. To determine if this mechanism of action contributes significantly to the therapeutic efficacy of AGI in the elderly, 10 type 2 diabetic subjects over the age of 65 years were given a standardized test meal with or without 25, 50, or 100 mg acarbose. The serum glucose, insulin, triglycerides and GLP-1 levels were measured at baseline and at 1 and 2 h postprandially. The anti-hyperglycemic effect of acarbose was maximal at 25-mg dose under these experimental conditions. Serum postprandial insulin and triglycerides levels were not significantly altered with acarbose treatment. The postprandial serum GLP-1 levels rose significantly only in two subjects and only during treatment with 100-mg acarbose. There were no significant correlations between serum GLP-1 and serum glucose or insulin levels. It is concluded that in most elderly type 2 diabetic subjects, maximal anti-hyperglycemic effects can be achieved with relatively small doses of acarbose and that GLP-1 is unlikely to contribute to the clinical efficacy of this agent in this subgroup of subjects. PMID- 11891019 TI - Combined aerobic and resistance exercise improves glycemic control and fitness in type 2 diabetes. AB - We investigated the effect of an 8 week circuit training (CT) program, combining aerobic and resistance exercise, on indices of glycemic control, cardiorespiratory fitness, muscular strength and body composition in 16 subjects (age 52 +/- 2 years) with type 2 diabetes using a prospective randomised crossover protocol. Submaximal exercise heart rate and rate pressure product were significantly lower after training (P<0.05), whilst ventilatory threshold increased (11.8 +/- 0.7 vs 13.8 +/- 0.6 ml kg(-1)min(-1), P<0.001). Muscular strength also increased with training (403 +/- 30 to 456 +/- 31 kg, P<0.001), whilst skinfolds (148.7 +/- 11.5 vs 141.1 +/- 10.7 mm, P<0.05), % body fat (29.5 +/- 1.0 vs 28.7 +/- 1.1%, P<0.05) and waist:hip ratio (99.2 +/- 1.5 vs 97.9 +/- 1.4%, P<0.05) significantly decreased. Concurrently, peak oxygen uptake (P<0.05) and exercise test duration (P<0.001) increased following training, whilst glycated hemoglobin (P<0.05) and fasting blood glucose (P<0.05) decreased. CT is an effective method of training that improved functional capacity, lean body mass, strength and glycemic control in subjects with type 2 diabetes. PMID- 11891018 TI - First human trial of pancreatic islet allo-transplantation in Korea--focus on re transplantation. AB - Over the past 20 years, allo-transplantation of islet or whole pancreas for reaching and sustaining near-normoglycemia, as close as possible to the physiological model, have been undertaken. As previously known, even though islet transplantation is possible as a safe re-transplant, it is not well known whether re-transplantation of islets is suitable for patients who have lost the grafted islet function. We have performed a human islet allo-transplantation and re transplantation on an IDDM patient for the first time in Asia and Korea. The recipient was a 32-year-old male and his insulin requirement was 75-85 U per day. After islet transplantation, the basal C-peptide increased from 0.6 to 2.1 ng/ml and insulin requirement decreased from 80 to 36 U per day, indicating that the grafted islets were functional. However, the grafted islets lost function 70 days after the transplantation. So, we performed re-transplantation of the islets. After the re-transplantation, the glucose profile became more stable and frequent episodes of severe hypoglycemia completely disappeared. His severe neuropathic pain improved dramatically and he could engage his ordinary daily life without any antineuropathic drugs. The success of this re-transplantation is one step closer to becoming a viable alternative for the millions of individuals who are suffering from diabetes. PMID- 11891020 TI - Retinopathy in a Chinese population with type 2 diabetes: factors affecting the presence of this complication at diagnosis of diabetes. AB - This study examined the prevalence of retinopathy in 2131 patients with type 2 diabetes attending a Beijing hospital for the first time. The median age of patients was 58 years (IQR 50-65). The overall prevalence of retinopathy was 27.3% (95% CI: 25.4-29.2) and for proliferative retinopathy 7.8% (95% CI: 6.7 8.9). When all patients were considered together, duration of diabetes (OR=1.8; P=0.001) and albumin excretion rate (OR=1.5; P=0.019) were independent risk factors for retinopathy. Blue-collar occupation (OR=1.5; P=0.047) and blood pressure (OR=1.2; P=0.021) were additional risk factors for non-proliferative and proliferative retinopathy respectively. Amongst the 773 newly diagnosed patients, 21% (95% CI: 17.8-23.6) already had retinopathy. The median age of those patients with retinopathy at diagnosis of diabetes was 3 years higher that those without retinopathy, and blue-collar workers (OR=2.2; P=0.012) as well as female gender were particularly at risk (OR=2.0; P=0.033). There was a strong correlation between duration of diabetes with the presence of retinopathy (r=0.95; P=0.01). By extrapolation, it could be estimated that some degree of hyperglycaemia might have been present for more than 20 years before diabetes was diagnosed. These findings emphasise the importance of earlier diagnosis of diabetes and its complications, especially in socially disadvantaged groups. PMID- 11891021 TI - Background factors correlated with the psychological features of 254 outpatients with Type 2 diabetes mellitus in Japan. AB - For the purpose of identifying the features of psychological troubles and their significance in Type 2 diabetes mellitus outpatients, we analyzed how psychological troubles were affected by various background factors. SUBJECTS AND METHODS The subjects all consisted of outpatients > or = 40 years of age at the Fukuoka Red Cross Hospital in December 1996. We used the State-Trait Anxiety Inventory (STAI) to determine anxiety, the Self rating Depression Scale to determine depression. We divided the patients into the ones who demonstrated each specific psychological trouble and the ones who did not, and then analyzed the psychological trouble between the two groups with correlation to various background factors which may have led the patients to develop their respective psychological features. RESULTS: In a stepwise multiple logistic regression analysis, the presence of itching and polyuria, age (40-49 years old) and females were correlated with anxiety in this study. Depression was also correlated with the absence of photo-coagulation therapy and the absence of leisure time activities. CONCLUSION: Our results identified some of the features of psychological troubles and their significance in diabetic outpatients, It is therefore important to carefully consider these factors during medical consultations. PMID- 11891023 TI - Calpain 10 gene polymorphisms are related, not to type 2 diabetes, but to increased serum cholesterol in Japanese. AB - A G-to-A (UCSNP-43) polymorphism of the calpain-10 gene was significantly associated with type 2 diabetes (DM) in Mexican-American, and was postulated, together with a T-to-C (UCSNP-44) polymorphism, as a risk factor for DM. We examined the association of these genotypes with DM in Japanese. Eighty-one subjects with DM and 81 non-diabetic subjects (NGT) were recruited. The number of subjects with genotypes UCSNP-43 G/G, G/A and A/A were 76, 5 and 0, respectively, for the DM and NGT groups. The number of subjects with genotypes UCSNP-44 T/T, T/C and C/C were 66, 14 and 1 for the DM group and 64, 17 and 0 for the NGT group. There was no difference between the groups in terms of frequency of any genotype combinations. No association between the genotypes and DM was observed. We next examined the differences between the genotypes or genotype combinations in terms of the traits related to DM, obesity, hypertension and dyslipidemia. No differences were observed between the genotypes UCSNP43 G/G and G/A, between UCSNP-44 T/T and the others, or between the genotype combination UCSNP-43 G/G and UCSNP-44 T/T and the others, except that the individuals with the genotype combination had significantly increased serum cholesterol levels (212.6 +/- 34.3 vs. 198.5 +/- 29.9, P=0.020). The genotype combination might be a risk factor, not for DM, obesity and hypertension, but for increased serum cholesterol. PMID- 11891022 TI - Relationship of the tumor necrosis factor-alpha -308 A/G promoter polymorphism with insulin sensitivity and abdominal fat distribution in Japanese patients with type 2 diabetes mellitus. AB - We investigated the relationship of the A/G variant of the tumor necrosis factor alpha (TNF-alpha) gene promoter at position -308 with insulin resistance and abdominal fat distribution in type 2 diabetic patients in the Japanese population. The TNF-alpha polymorphism was evaluated by polymerase chain reaction restriction fragment length polymorphism in 142 healthy volunteers and 132 type 2 diabetic patients. Insulin sensitivity was assessed by homeostasis model assessment (HOMA) index in healthy subjects and hyperinsulinemic euglycemic clamp in type 2 diabetic patients. Abdominal fat distribution was evaluated by computed tomography (CT) scanning in diabetic patients. The TNF-alpha polymorphism was detected in three healthy volunteers and three type 2 diabetic patients, all of them being heterozygotes. There was no significant difference in allele frequencies of the -308 polymorphism between healthy subjects (0.0106) and type 2 diabetic patients (0.0114). HOMA index was no significant difference between healthy subjects with and without polymorphism (1.09 +/- 0.03 vs. 1.02 +/- 0.05). Glucose infusion rate (GIR), an index of insulin sensitivity, was not significantly different between diabetic patients with and without TNF-alpha polymorphism (40.4 +/- 4.1 vs. 45.0 +/- 1.8 micromol/kg per min). Moreover, no remarkable effect of TNF-alpha polymorphism on abdominal fat distribution was observed in diabetic patients. These results suggest that A/G heterozygotes of the TNF-alpha gene promoter at position -308 play no major role in the pathogenesis of insulin resistance or abdominal fat distribution in Japanese type 2 diabetic patients. PMID- 11891024 TI - Incidence and seasonal variation of Type 1 diabetes in children in Farwania area, Kuwait (1995-1999). AB - The aim of our study was to confirm the continuing rise in the incidence of Type 1 diabetes among Kuwaiti children aged 0-14, and to assess the effect of seasonality on incidence. Data from all newly diagnosed diabetic children between the period of 1995 and 1999 were analyzed. A total of 129 cases of Type 1 diabetes were diagnosed during the study period, of whom 68 were Kuwaiti nationals and were included in the study. The incidence was 20.18 per 100,000 (95% CI 16.3-28.2). Incidence rates for the age-groups 0-4, 5-9 and 10-14 were 8.12, 21.07 and 34.06, respectively. There was a significant female predominance (F:M ratio was 1.4:1, P<0.05). More cases were diagnosed in the cool months (November-February) compared with the warm months (June-September, P<0.05). There was increase in incidence from 1995 to 1999, but compared with data from the 1980s on the same age group, incidence has increased. A positive family history of Type 1 diabetes in a close relative was recorded for 30% of the patients. Although, only data from one hospital were included, Kuwait is very small geographically and not likely to have differences between different areas. Stress factors, economic growth, changes in the nutritional habits and the adoption of the western lifestyle may explain some of this increase. PMID- 11891025 TI - A randomized trial of systematic nodal dissection in resectable non-small cell lung cancer. AB - PURPOSE: We conducted a randomized trial to investigate whether systematic nodal dissection (SND) is superior to mediastinal lymph nodal sampling (MLS) in surgical treatment of non-small cell lung cancer (NSCLC). METHODS: The patients resectable clinical Stage I-IIIA NSCLC were randomly assigned to lung resection combined with SND or lung resection combined with MLS. After postoperative pathological re-staging, eligible cases were followed up until 30 November 2000. The Kaplan-Meier method was used for survival analysis. COX proportional hazards model was used for prognostic analysis. RESULTS: Of the 532 patients who were enrolled in the study, 268 patients were assigned to lung resection combined with SND and 264 were assigned to lung resection combined with MLS. After surgical restaging only 471 cases were eligible for follow-up. The median survival was 59 months in the group given SND and 34 months in the group given MLS (P=0.0000 by the log rank test). There was significant difference in survival in Stage I (5 year survival 82.16 vs. 57.49%) and Stage IIIA (26.98 vs. 6.18%) by the log rank test and Breslow test. There was no significant yet marginal difference in survival by log rank test (10-year survival 32.04 vs. 26.92%, P=0.0523) but significant difference in survival by Breslow test (5-year survival 50.42 vs. 34.05%, P=0.0284) in Stage II. Types of mediastinal lymph node dissection, pTNM stage, tumor size and number of lymph node metastasis were four factors that influenced long-term survival rate by multivariate analysis. CONCLUSIONS: As compared with MLS, lobectomy (pneumonectomy) combined with SND can improve survival in resectable NSCLC. PMID- 11891026 TI - Complete mediastinal lymph node dissection---does it make a difference? PMID- 11891027 TI - A prospective study of a total material of lung cancer from a county in Sweden 1997-1999: gender, symptoms, type, stage, and smoking habits. AB - The epidemiology of lung cancer is changing in many parts of the world. In the industrialized countries, there is a trend that the incidence in men is declining, while it is increasing for women. Also, adenocarcinomas are becoming relatively more common, especially among men. The purpose of this study was to investigate whether such trends also occur in Sweden and also to describe other aspects of an unselected lung cancer material today, such as symptoms, stage and smoking habits. In the county of Gaevleborg, Sweden, practically all patients with lung cancer are referred to the lung department, and thus a total material of lung cancer patients with only a minimal selection bias can be studied. All patients with lung cancer in the county from January 1, 1997 to December 31, 1999, were investigated prospectively regarding stage, type of cancer, and symptoms. In all, there were 364 patients, 237 (65.1%) men and 127 (34.9%) women. The mean age for men was 69.8 and for women, 68.1 years. 91.9% of the men and 78.6% of the women were smokers or ex-smokers. In general the men were heavier smokers than were the women (P<0.0001). Adenocarcinoma was the most common subtype found in women and squamous cell carcinoma in men. The excess of adenocarcinoma in women was due to never-smoking women; for smoking and ex smoking men and women, the proportion of adenocarcinomas was the same. In all, 240 patients (68.0%) were diagnosed at Stage IIIb (27.2%) or IV (40.8%), with no significant differences between the sexes. The most common first symptom was cough. Only 7.0% of patients were asymptomatic. In conclusion, the trend of an increasing proportion of adenocarcinoma in lung cancer is seen also in Sweden. A depressingly high percentage of patients present in late stages and are thus inoperable. PMID- 11891028 TI - Lys751Gln polymorphism in the DNA repair gene XPD and risk of primary lung cancer. PMID- 11891029 TI - Modulation of tumor-infiltrating lymphocyte cytolytic activity against human non small cell lung cancer. AB - The cytokines expressed in tumor microenvironments are thought to be important mediators of both the host immune response and tumor survival. The source of these cytokines includes tumor cells, infiltrating leukocytes, fibroblasts, and other stromal elements. We previously reported that tumor-infiltrating lymphocytes (TIL) from human non-small cell lung cancer (NSCLC) express predominantly type 1 cytokines, which are known to enhance cell-mediated immunity. The purpose of this study is to assess the cytokine mRNA expression of human NSCLC primary cell lines and the capacity of the tumor-associated cytokines to modulate the development of TIL cytolytic activity against the autologous tumor. Cytokine mRNA expression was determined by RT-PCR and the capacity of TIL to kill autologous lung tumor cells was measured by the chromium-51 (51Cr) release assay. All NSCLC primary cell lines expressed mRNA for IL-4, IL-6, and transforming growth factor-beta1 (TGFbeta1), whereas IL-10 was expressed in only 1/7 cell lines. When added to TIL cultures stimulated with anti-CD3+IL-2, IL-4 and IL-10 enhanced and TGF-beta1 suppressed the development of TIL cytolytic activity against autologous tumor cells. The effects of IL-6 were inconsistent and for the group, were not statistically significant. These results demonstrate that human NSCLC cells express cytokines with the capacity to regulate the in situ anti-tumor immune response. However, the effects of tumor-derived cytokines varied qualitatively and quantitatively suggesting the balance between specific type 2 cytokines or TGF-beta1 within tumor microenvironments may influence prognosis or response to immunotherapy. PMID- 11891030 TI - Apoptotic tumor-cell death in response to cell proliferation is influenced by p53 status in resected non-small cell lung cancer. AB - To evaluate the influence of p53 status on postoperative survival and incidence of apoptosis (apoptotic index, AI) in non-small cell lung cancer (NSCLC), a total of 185 pathologic stage I patients were retrospectively analyzed. It was demonstrated by univariate and multivariate analyses that aberrant expression of p53 was a significant factor to predict a poor prognosis, which was caused by a significantly higher proliferative index (PI) in tumor with aberrant expression of p53 (52.7%) than that in tumor without aberrant expression of p53 (37.9%, P < 0.001). In addition, for tumor without aberrant expression of p53, mean AIs of the lowest-, the lower-, the higher-, and the highest-PI groups were 12.6, 12.9, 31.3, and 35.1, respectively, showing that incidence of apoptosis was markedly increased in response to cell proliferation (P = 0.002). In contrast, for tumor with aberrant expression of p53, no increase in incidence of apoptosis in response to cell proliferation was documented. These results clearly demonstrated that active cell proliferation and reduced apoptotic cell death in response to cell proliferation resulted in the poor postoperative prognosis in tumor with aberrant expression of p53. PMID- 11891031 TI - AH6809 antagonizes non-small cell lung cancer prostaglandin receptors. AB - The effects of prostaglandin (PG)E2 on lung cancer cells were investigated. 3H PGE2 bound with high affinity to membranes derived from small cell lung cancer (SCLC) and non-SCLC (NSCLC) cell lines. Using NSCLC NCI-H1299 membranes, specific 3H-PGE2 binding to NCI-H1299 membranes was inhibited with moderate affinity by PGE2, PGE1, PGF2alpha and 6-isopropoxy-9-xanthone-2-carboxylic acid (AH6809) but not PGD2, LTB4 or 5-HETE. By RT-PCR, EP2 receptor PCR products were detected in extracts derived from lung cancer cells. PGE2 caused cAMP elevation in a concentration-dependent manner using NCI-H1299 cells and the increase in cAMP caused by PGE2 was antagonized by AH6809. PGE2 had no effect on cytosolic Ca2+ but PGE2 caused increased c-fos mRNA in NCI-H1299 cells. AH6809 inhibited the proliferation of NCI-H1299 cells using MTT and clonogenic assays. These results indicate that functional PG receptors are present on NSCLC cells which are antagonized by AH6809. PMID- 11891033 TI - Radiologic-prognostic correlation in patients with small pulmonary adenocarcinomas. AB - We retrospectively studied the correlation between high-resolution computed tomography (HRCT) findings and both pathologic findings and prognosis in 137 patients with small pulmonary adenocarcinomas (tumor diameter 20 mm or less). We then evaluated the usefulness of our classification on HRCT findings for predicting outcome and formulating a therapeutic strategy. Tumors were classified according to attenuation of on HRCT images as the 'air-containing type' or the 'solid-density type'. The air-containing type adenocarcinomas (n = 66) revealed no lymphnode involvement detected surgically or pathologically, and revealed minimal microscopic evidence of metastasis (pleural involvement, vascular invasion, or lymphatic permeation). Furthermore, the relapse-free and overall survival curves of the air-containing types revealed no relapses or deaths. Patients with solid-density type adenocarcinomas (n = 71), on the other hand, demonstrated a poor prognosis. Consequently, it seems likely that air-containing adenocarcinomas 20 mm or less in diameter on HRCT may be treated successfully by limited lung resection. PMID- 11891032 TI - Survival and risk model for stage IB non-small cell lung cancer. AB - BACKGROUND: The aim of this work is to estimate the prognostic value of a set of clinical-pathological factors in patients resected for non-small cell lung cancer (NSCLC) and classified as stage IB, in order to create a prognostic model for establishing risk groups, and to validate that model. METHODS: Among 637 patients resected and classified as stage IB, we analyzed sex, age, symptoms, location, type of resection, cell type, histology, and tumor size. The Kaplan-Meier method was used to estimate the survival. The results were compared using the log-rank test. All the significant variables from this univariable method were then included in a multivariable method of estimation of the proportional risk for survival data developed by Cox, using the variables selected, a regression model was developed for accurately predicting survival. To validate the predictive capability of the regression model, we randomly divided our patients into training and test subsets, containing 322 and 315 cases, respectively. RESULTS: The overall 5-year survival rate of the series was 60%. The cell type, the squamous or non-squamous and the tumor size showed a significant influence on survival in the univariable analysis, while, according to the Cox model, only the tumor size and the squamous or non-squamous type entered into regression. Hazard rates were calculated for each patient. The mean risk was 0.87 +/- 0.25 (range 30 1.94). The series was divided into three risk groups (low, intermediate, and high risk) according to the fitted hazard rates, using cut-off points (one standard deviation from the mean). The 5-year survival rates were 85, 59, and 44%, respectively. To validate the model, we repeated the analysis for training and test subsets. Only the tumor size had a significant influence on survival in the univariable analysis. Using the Cox model, also the tumor size entered into regression. The mean risk was 0.79 +/- 0.29 (range 0.09-2.12). Cut-off points were 0.50 and 1.08 for the low, intermediate, and high-risk groups. The 5-year survival rates were 83, 58, and 40%, respectively. We validated the regression model obtained in the training subset by demonstrating its capacity in identifying risk groups in the test subset. The 5-year survival rates were 83, 61, and 49.5% for the low, intermediate, and high-risk groups, respectively (P = 0.0104). CONCLUSIONS: Stage IB does not succeed in configuring a group of patients with a homogeneous prognosis, as there is a wide variability in a 5-year survival. The estimation of prognosis derived from a multivariable analysis can obviate the limitations of the actual staging system for NSCLC. PMID- 11891034 TI - Influence of delays on survival in the surgical treatment of bronchogenic carcinoma. AB - The objective of the study was to find out whether the delay in time from when bronchogenic carcinoma is diagnosed until a therapeutic thoracotomy is performed affects patient survival. The population analysed comprised 1082 patients with clinical stage I and II, non-small cell lung cancer (NSCLC), who had been operated on between October 1993 and September 1997, and were registered in the Bronchogenic Carcinoma Cooperative Group of the Spanish Society of Pneumology and Thoracic Surgery (GCCB-S). In this series, the median therapeutic delay was 35 days (1-154), with a median survival rate of 52 months (45.6-58.3). A statistical study was developed that, in addition to the delay, included the variables of age, histology, clinical stage, and pathological stage of the disease. Therapeutic delay was included in the multivariable analysis as a quantitative and qualitative variable and a comparison among the different intervals of delay in days (1-20 vs. 21-40 vs. 41-60 vs. > 60) was performed in order to ascertain its influence on survival. Univariate and multivariate Cox's regression analyses showed that age (> 70 years), clinical stage (I vs. II), and pathological stage influence survival. As for the histology and the delay, no significant differences were observed in the survival of any of the intervals even when compared against the intervals at the extremes (1-20 vs. > 60). In conclusion we found no influence of delay upon the survival. PMID- 11891035 TI - Prognosis of non-surgically treated, clinical stage I lung cancer patients in Japan. AB - OBJECTIVE: The optimal management of stage I lung cancer is surgical resection. However, some of these patients are not candidates for surgery because of several medical problems. We analyzed prognosis of non-surgically treated, clinical stage I lung cancer patients. METHODS AND RESULTS: There were 21211 lung cancer patients registered from 1982 to 1991 in the data-base of the Japanese National Chest Hospital Study Group for Lung Cancer, and the number of non-surgically treated, clinical stage I lung cancer patients during the 10 years was 802. The 5 and 10-year survival rates of the 799 patients, exclusive of two carcinoid tumors and one adenid cystic carcinoma which have good prognosis, were 16.6 and 7.4%. We analyzed the 799 patients according to several prognostic factors. Sex, T factor of the tumor, histology, performance status and the method in which lung cancer was detected were prognostic factors, but age and treatment method were not associated with prognosis. Forty-nine patients survived for 5 years or more without surgical resection, but the survival rate continued to decrease even after 5 years, and the 7- and 10- year survival rates were 34.4 and 18.1% in the 49 patients. CONCLUSIONS: It is a fact that there are long-term survivors in non surgically treated, stage I lung cancer patients. However, the rate is low, and the survival curve continues to decrease even after 5 years. Long-term survivors might suggest the presence of a lung cancer in which the tumor growth is slow. PMID- 11891036 TI - Decreased perioxisome proliferator-activated receptor gamma gene expression was correlated with poor prognosis in patients with lung cancer. AB - Activation of the nuclear hormone receptor perioxisome proliferator-activated receptor gamma (PPARgamma) inhibits cell growth and induces apoptosis in several human cancers. We have hypothesized that PPARgamma mRNA levels could be predictors of the differentiation and survival of lung cancer. The study included 77 lung cancer cases. The mRNA levels were quantified by real time reverse transcription-polymerase chain reaction (RT-PCR) using LightCycler. The PPARgamma mRNA levels were decreased in tumor tissues from lung cancer (0.579 +/- 1.255) compared to the normal adjacent lung tissues (4.191 +/- 2.868) (P = 0.0001). No significant difference in PPARgamma mRNA levels was found among gender, age, and pathological subtype. The PPARgamma mRNA levels were higher in tumor tissues from higher differentiated lung cancer. The NSCLC patients with low PPARgamma mRNA expression (< 0.5) had significantly worse survival than the patients without low PPARgamma mRNA levels (P = 0.0438, Breslow-Gehan-Wilcoxon test; P = 0.0168, Cox's proportional-Hazards regression model). Thus, PPARgamma mRNA levels may serve as a prognostic marker in lung cancer. Using the LightCycler RT-PCR assay, the determination of PPARgamma mRNA levels might provide a potential marker for treatment of lung cancer by PPARgamma agonist. However, further studies and a longer follow up are needed to confirm the impact of PPARgamma in the biological behavior of the tumor. PMID- 11891037 TI - Distribution of talc suspension during treatment of malignant pleural effusion with talc pleurodesis. AB - Talc pleurodesis is an effective technique for the management of symptomatic malignant pleural effusions. It is assumed that a good dispersion of talc suspension contributes to the final success of this treatment. For this purpose, guidelines often advise to rotate the patient after intra-pleural instillation of the sclerosant. This prospective, randomized study analyses the dispersion of talc suspension and the overall success rate in patients with malignant effusions. After instillation of 99mTc-sestamibi-labeled talc suspension ten subjects were rotated for 1 h, while the ten other patients remained in a stable supine body position. Scintigraphic imaging was done in two directions immediately after instillation and after 1 h with a clamped drain. The overall success of the treatment was assessed 1 month after the pleurodesis. The dispersion of talc was limited and unequal in 75% of the subjects. In two patients with apparently good distribution on anterior views, the lateral views of the scintigraphy showed only limited distribution. Rotation of the patients did not influence the dispersion of sludge after 1 min or 1 h. Pleurodesis was successful in 85% of the patients after 1-month follow-up. Standard rotation protocols for patients with malignant pleural effusion do not affect the overall dispersion of talc suspension and should be abolished because of the discomfort caused to the patients. PMID- 11891038 TI - CEA and CA 549 in serum and pleural fluid of patients with pleural effusion. AB - BACKGROUND: The determination of the pleural fluid (PF) carcinoembryonic antigen (CEA) concentration has proved helpful in the differentiation between pleural effusions (PE) of malignant and benign origin. The present study was designed to prospectively compare the utility of CEA with that of a recently introduced tumour marker, carbohydrate antigen 549 (CA 549). PATIENTS AND METHODS: In 383 consecutive patients referred for thoracentesis (130 malignant and 253 benign), pleural and serum levels of CEA and CA 549 were, respectively, determined by enzyme immunoassay (EIA) and immunoradiometric assay (IRMA). RESULTS: CEA and CA 549 showed a high specificity for malignancy in serum (97 and 96%, respectively) and PF (98 and 99%). The serum sensitivity was 33% for CEA and 47% for CA 549 while in PF was 49 and 54%, respectively. The area under the curve of CA 549 (0.78) was significantly larger than that of CEA (0.66) in serum (P < 0.005) and in PF (0.83 and 0.75, respectively, P < 0.02) as well. CA 549 showed a higher sensitivity (P < 0.001) than CEA for ovarian tumours. In PF, the accuracy of the combination of both markers was higher than that of any individual marker, although the difference was only significant with respect to CEA (P < 0.02). CONCLUSIONS: The results of the present study show that a new tumour marker CA 549 is at least similar in terms of sensitivity and specificity to CEA in the evaluation of patients with PE of unknown cause. PMID- 11891039 TI - Low morbidity of bronchoplastic procedures after chemotherapy for lung cancer. AB - OBJECTIVE: To evaluate if induction chemotherapy, with or without irradiation, represents an additional risk factor for early and late morbidity and perioperative mortality in bronchoplastic procedures for lung cancers. METHODS: From January 1998 to January 2001, 27 patients underwent a bronchial sleeve resection after induction treatment at the European Institute of Oncology in Milan. They represent 7% of lung cancer resections (387) and 27% of those performed after neoadjuvant treatment (100 cases). Histology was: 17 epidermoid carcinoma, 8 adenocarcinoma and 2 SCLC. Twenty-four patients (89%) received a preoperative cisplatin based polichemotherapy, and three cases (11%) a chemo radiation therapy. A right sleeve lobectomy or bilobectomy was undertaken in 21 patients (78%) and a left lobectomy in 6 (22%). A resection of tracheal carina was associated in three cases and a vascular resection in 10 (five vena cava and five pulmonary artery). Twelve patients (44%) received adjuvant mediastinal irradiation. Perioperative morbidity of the study group (group 1) was compared with that of patients submitted to sleeve resection without neoadjuvant treatment (group 2), or standard pneumonectomy after induction treatment (group 3). RESULTS: There were no postoperative deaths. A major perioperative complication occurred in two patients (7%) of group 1, one patient of group 2 (3.5%), and four in group 3 (17%). Among patients of the study group, no anastomotic dehiscence or pleural empyema were observed. Only one late anastomotic stricture occurred after postoperative radiation treatment. No significant difference in early and late complication rate was found between the three groups of patients. High rate of complete resection was achieved (93%) in patients of the study group and extent of nodal dissection was similar between sleeve resections and pneumonectomy patients. CONCLUSIONS: Preoperative chemotherapy or combination of chemo-radio therapy is not associated with an additional risk of anastomotic complications in bronco and angioplastic procedures. Parenchyma sparing resection is a valid option for selected patients with locally advanced lung cancer after induction treatment. A longer follow up is necessary to evaluate efficacy of the procedure in term of survival and local control. PMID- 11891040 TI - Activity of gemcitabine and carboplatin in advanced non-small cell lung cancer: a phase II trial. AB - This phase II trial was designed to investigate the efficacy and tolerability of gemcitabine combined with carboplatin in patients with advanced or metastatic NSCLC. Patients were treated with gemcitabine 1000 mg/m(2) on days 1, 8, and 15 of a 28-day cycle and carboplatin AUC 5 mg/ml/min on day 2 of each cycle. Fifty patients (Zubrod-ECOG-WHO performance status 0/1 in 70/30%, stage IV disease in 64%) entered the study and were evaluable for response and toxicity. There was 1 complete response and 24 partial responses among 50 evaluable patients, for a response rate of 50% (95% CI: 36.0-64.1%). The median survival time was 13 months (range: 6-22 months), and the 1-year survival rate was 54%. Hematologic toxicities included grades 3 and 4 neutropenia in 24 and 8% of patients, respectively, and grades 3 and 4 thrombocytopenia in 48 and 8% of patients, respectively. These were without clinical sequelae. Seven (14%) patients had grade 3 nausea and vomiting. The combination of gemcitabine and carboplatin is highly active and well tolerated in patients with advanced or metastatic NSCLC. PMID- 11891041 TI - A phase II study of continuous concurrent thoracic radiotherapy in combination with mitomycin, vindesine and cisplatin in unresectable stage III non-small cell lung cancer. AB - The split-course concurrent thoracic radiation therapy (TRT) and full-dose chemotherapy for unresectable stage III non-small cell lung cancer (NSCLC) has produced promising results by comparison with the sequential approach. Instead of split-course radiation, we conducted a phase II study to investigate the feasibility of continuous concurrent TRT and chemotherapy. Twenty-two patients with unresectable NSCLC were enrolled onto a phase II study of continuous concurrent radiotherapy and chemotherapy. Treatment consisted of two courses of cisplatin (80 mg/m(2) on days 1 and 29), vindesine (3 mg/m(2) on days 1, 8, 29 and 36), and mitomycin (8 mg/m(2) on days 1 and 29). TRT began on day 2 at a dose of 60 Gy (2 Gy per fraction and 5 fractions per week for a total of 30 fractions). Of 22 patients assessable for response, none achieved a CR and 17 (77.3%) achieved a PR with an overall response rate of 77.3% (95% confidence interval, 54.6-92.2%). Grade 3 or 4 leukopenia was observed in 5/13 (81.8%) patients. Six patients (27.3%) experienced > or = grade 3 thrombocytopenia. Non hematological toxicity was relatively mild. The overall median survival time was 19.0 months and the 1- and 2-year survival rates were 84.8 and 34.5%, respectively. It was possible to administer two courses of chemotherapy in 18 patients (81.8%) as planned. Nineteen (86.4%) of the 22 patients received the planned 60 Gy radiation. It seems to be difficult to administer the planned treatment without any interruption for the majority of patients. However, in the selected patients who completed the 60 Gy TRT, nearly half of the patients completed TRT without interruption. This combination regimen is considered to be feasible on condition that the stopping rule of the treatment is followed. We recommend administering radiotherapy continuously as far as possible. PMID- 11891042 TI - Transcriptional mechanisms regulating myeloid-specific genes. AB - Myeloid blood cells comprise an important component of the immune system. Proper control of both lineage- and stage-specific gene expression is required for normal myeloid cell development and function. In recent years, a relatively small number of critical transcriptional regulators have been identified that serve important roles both in myeloid cell development and regulation of lineage restricted gene expression in mature myeloid cells. This review summarizes our current understanding of the regulation of lineage- and stage-restricted transcription during myeloid cell differentiation, how critical transcriptional regulators control myeloid cell development, and how perturbations in transcription factor function results in the development of leukemia. PMID- 11891043 TI - Human homologue of a gene mutated in the slow Wallerian degeneration (C57BL/Wld(s)) mouse. AB - The slow Wallerian degeneration mouse (C57BL/Wld(s)) is a mutant strain of mouse, with the unique phenotype of prolonged survival of the distal axon following axotomy. The causative mutation is an 85 kb tandem triplication on distal mouse chromosome 4. The dominant slow Wallerian degeneration phenotype is conferred by a hybrid gene within the triplication, comprising a gene of previously unknown function, D4Cole1e, and the 5' end of ubiquitination factor E4B (Ube4b). It encodes an in-frame fusion protein consisting of the N-terminal 70 amino acids of Ube4b and 303 amino acids derived from the D4Cole1e gene. We have identified the human homologue of D4Cole1e, and mapped it to chromosome 1p36.2. Additional fluorescence in situ hybridisation signals indicate the presence of several homologous human sequences. Northern blot analysis shows two transcripts, widely expressed at varying levels in different human tissues. The human cDNA, which encodes a protein of 279 amino acids, has 80% nucleotide identity with the mouse cDNA. The derived human and mouse protein sequences share 78% amino acid identity and 82% amino acid similarity. The human cDNA and protein sequences are identical to the human nicotinamide mononucleotide adenylyltransferase (NMNAT). We have also determined the intron/exon structure of the gene, which will facilitate the screening of these exons for mutations in human neurodegenerative disorders. PMID- 11891044 TI - Ermelin, an endoplasmic reticulum transmembrane protein, contains the novel HELP domain conserved in eukaryotes. AB - We have cloned a cDNA encoding a novel protein referred to as ermelin from mouse C2 skeletal muscle cells. This protein contained six hydrophobic amino acid stretches corresponding to transmembrane domains, two histidine-rich sequences, and a sequence homologous to the fusion peptides of certain fusion proteins. Ermelin also contained a novel modular sequence, designated as HELP domain, which was highly conserved among eukaryotes, from yeast to higher plants and animals. All these HELP domain-containing proteins, including mouse KE4, Drosophila Catsup, and Arabidopsis IAR1, possessed multipass transmembrane domains and histidine-rich sequences. Ermelin was predominantly expressed in brain and testis, and induced during neuronal differentiation of N1E-115 neuroblastoma cells but downregulated during myogenic differentiation of C2 cells. The mRNA was accumulated in hippocampus and cerebellum of brain and central areas of seminiferous tubules in testis. Epitope-tagging experiments located ermelin and KE4 to a network structure throughout the cytoplasm. Staining with the fluorescent dye DiOC(6)(3) identified this structure as the endoplasmic reticulum. These results suggest that at least some, if not all, of the HELP domain-containing proteins are multipass endoplasmic reticulum membrane proteins with functions conserved among eukaryotes. PMID- 11891045 TI - Genomic organization and differential expression of Kalirin isoforms. AB - Multidomain guanine nucleotide (GDP/GTP) exchange factor (GEF) proteins coordinate diverse inputs that signal the actin cytoskeleton. Mammals have two such proteins (Kalirin, Trio), while Drosophila has one, which plays essential roles within and outside the nervous system. For Kalirin, numerous isoforms containing different combinations of functional domains are generated through alternative splicing and use of alternative transcriptional start sites. These different isoforms potentially allow a wide variety of proteins to interact with Kalirin, thereby affecting the activity of the functional domains. Humans, like rats, express a large set of Kalirin isoform mRNAs, and we identified a novel Kalirin isoform, containing only the second GEF domain. Kalirin isoforms are predominantly expressed in the brain, while Trio is expressed in a wider variety of tissues. Alternative splicing and transcription of Kalirin are differentially regulated during development in rats and humans, resulting in expression of isoforms of Kalirin containing different functional domains at different times and locations. The prevalence of Kalirin in the cortex throughout life suggests roles in axonal development and the mature brain. PMID- 11891046 TI - Genomic structure of newly identified paralogue of RNA helicase II/Gu: detection of pseudogenes and multiple alternatively spliced mRNAs. AB - RNA helicase II/Gu (RH-II/Gu or DDX21) is a DEAD-box enzyme that localizes to the nucleoli and may be involved in ribosomal RNA synthesis or processing. It has two paralogues, RH-II/Gualpha and RH-II/Gubeta, both genes of which are on chromosome 10. Their similar genomic structures suggest the two genes arose by gene duplication. Both genes are expressed at higher levels in some normal human tissues compared to matching tumor tissues. Pseudogenes for RH-II/Gubeta exist on chromosomes 2, 3 and 4. No pseudogene was identified for RH-II/Gualpha. Both exon inclusion and exon skipping were found to post-transcriptionally regulate RH II/Gubeta gene expression. No alternative splicing was identified for RH II/Gualpha. Overall, the results suggest that the two paralogues of RH-II/Gu arose by gene duplication but the resulting genes are differentially regulated. PMID- 11891047 TI - Allelic variation in the highly polymorphic locus pspC of Streptococcus pneumoniae. AB - PspC, also called SpsA, CbpA, PbcA, and Hic, is a surface protein of Streptococcus pneumoniae studied for its antigenic properties, its capability to bind secretory IgA, C3 and complement factor H, and its activity as an adhesin. In this work we characterized the pspC locus of 43 pneumococcal strains by DNA sequencing of PCR fragments. Using PCR primers designed on two unrelated open reading frames, flanking the pspC locus, it was possible to amplify the pspC locus of each of the 43 strains of S. pneumoniae. In 37 out of 43 strains there was a single copy of the pspC gene, while two tandem copies of pspC were found in the other six strains. The sequence of the pspC locus was different in each of the 43 strains. Insertion sequences were found in the pspC locus of 11 out of 43 strains. Analysis of the deduced amino acid sequence of the PspC variants showed a common organization of the molecules: (i) a 37 amino acid leader peptide which is conserved in all proteins, (ii) an N-terminal portion which is essentially alpha-helical, and is the result of assembly of eight major sequence blocks, (iii) a proline-rich region, and (iv) a C-terminal anchor responsible for the cell surface attachment. By sequence comparison we identified 11 major groups of PspC proteins. Proteins within one group displayed only minor variations of the amino acid sequence. An unexpected finding was that PspC variants could differ in the anchor sequence. While 32 of the PspC proteins displayed the typical choline binding domain of pneumococcal surface proteins, 17 other PspCs showed the LPXTG motif, which is typical of surface proteins of other gram-positive bacteria. This major difference in the anchor region was also observed in the adjacent proline rich regions which differed considerably in size and composition. PMID- 11891048 TI - MLL3, a new human member of the TRX/MLL gene family, maps to 7q36, a chromosome region frequently deleted in myeloid leukaemia. AB - We characterized MLL3, a new human member of the TRX/MLL gene family. MLL3 is expressed in peripheral blood, placenta, pancreas, testes, and foetal thymus and is weakly expressed in heart, brain, lung, liver, and kidney. It encodes a predicted protein of 4911 amino acids containing two plant homeo domains (PHD), an ATPase alpha_beta signature, a high mobility group, a SET (Suppressor of variegation, Enhancer of zeste, Trithorax) and two FY (phenylalanine tyrosine) rich domains. The amino acid sequence of the SET domain was used to obtain a phylogenetic tree of human MLL genes and their homologues in different species. MLL3 is closely related to human MLL2, Fugu mll2, a Caenorhabditis elegans predicted protein, and Drosophila trithorax-related protein. Interestingly, PHD and SET domains are frequently found in proteins encoded by genes that are rearranged in different haematological malignancies and MLL3 maps to 7q36, a chromosome region that is frequently deleted in myeloid disorders. Partial duplications of the MLL3 gene are found in the juxtacentromeric region of chromosomes 1, 2, 13, and 21. PMID- 11891049 TI - Phylogenetic position of Rhynchopus sp. and Diplonema ambulator as indicated by analyses of euglenozoan small subunit ribosomal DNA. AB - The taxa Rhynchopus Skuja and Diplonema Griessmann were first described as remarkable protists with euglenid affinities. Later on, the placement of Diplonema within the Euglenozoa was confirmed by molecular data. For this study two new sequences were added to the euglenozoan data set. The uncertainly placed Rhynchopus can be identified as a close relative to Diplonema by small subunit ribosomal DNA (SSU rDNA) analysis. The new sequence of Diplonema ambulator is in close relationship to two other Diplonema species. Our molecular analyses clearly support the monophyly of the diplonemids comprising Rhynchopus and Diplonema. Yet the topology at the base of the euglenozoan tree remains unresolved, and especially the monophyly of the euglenids is arguable. SSU rDNA sequence analyses suggest that significantly different GC contents, high mutational saturation in the euglenids, and different evolutionary rates in the euglenozoan clades make it difficult to identify any sister group to the diplonemids. PMID- 11891050 TI - Gonad-inhibiting hormone of the Norway lobster (Nephrops norvegicus): cDNA cloning, expression, recombinant protein production, and immunolocalization. AB - The gonad-inhibiting hormone (GIH) belongs to a neuropeptide family synthesized and released in a neurohemal complex of crustacean eyestalks. The GIH is involved in gonad maturation and plays a more complex role in the control of reproduction and molting. With a combination of reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends approaches we determined the cDNA sequence of the Norway lobster Nephrops norvegicus prepro GIH. The open reading frame of 339 bp codes for a polypeptide of 112 amino acids showing 96% identity with the other known GIH of Homarus americanus. The precursor peptide consists of a putative signal peptide of 31 amino acids and a putative mature peptide region of 81. RT-PCR analysis shows that GIH mRNA is expressed mainly in eyestalks, both in female and male; the expression of GIH mRNA also in supraesophageal ganglia suggests the existence of additional GIH-producing neurons besides those of eyestalks. A specific polyclonal antibody was raised against a portion of the mature peptide region obtained through expression in Escherichia coli fused to glutathione-S-transferase. Immunocytochemical studies were carried out by using this antibody in N. norvegicus and in other crustaceans, Munida rugosa and Squilla mantis; these locate GIH in superficial axon terminals of the releasing organ, the sinus gland. The identification of a second GIH sequence in crustaceans allows to hypothesize the occurrence, within the neuropeptide family, of three subfamilies probably involved in different functions: crustacean hyperglycemic hormones, GIHs and molt-inhibiting hormones/mandibular organ-inhibiting hormones. PMID- 11891051 TI - A hydrogenosomal [Fe]-hydrogenase from the anaerobic chytrid Neocallimastix sp. L2. AB - The presence of a [Fe]-hydrogenase in the hydrogenosomes of the anaerobic chytridiomycete fungus Neocallimastix sp. L2 has been demonstrated by immunocytochemistry, subcellular fractionation, Western-blotting and measurements of hydrogenase activity in the presence of various concentrations of carbon monoxide (CO). Since the hydrogenosomal hydrogenase activity can be inhibited nearly completely by low concentrations of CO, it is likely that the [Fe] hydrogenase is responsible for at least 90% of the hydrogen production in isolated hydrogenosomes. Most likely, this hydrogenase is encoded by the gene hydL2 that exhibits all the motifs that are characteristic of [Fe]-hydrogenases. The open reading frame starts with an N-terminal extension of 38 amino acids that has the potential to function as a hydrogenosomal targeting signal. The downstream sequences encode an enzyme of a calculated molecular mass of 66.4 kDa that perfectly matches the molecular mass of the mature hydrogenase in the hydrogenosome. Phylogenetic analysis revealed that the hydrogenase of Neocallimastix sp. L2. clusters together with similar ('long-type') [Fe] hydrogenases from Trichomonas vaginalis, Nyctotherus ovalis, Desulfovibrio vulgaris and Thermotoga maritima. Phylogenetic analysis based on the H-cluster - the only module of [Fe]-hydrogenases that is shared by all types of [Fe] hydrogenases and hydrogenase-like proteins - revealed a monophyly of all hydrogenase-like proteins of the aerobic eukaryotes. Our analysis suggests that the evolution of the various [Fe]-hydrogenases and hydrogenase-like proteins occurred by a differential loss of Fe-S clusters in the N-terminal part of the [Fe]-hydrogenase. PMID- 11891052 TI - An Agrobacterium gene involved in tumorigenesis encodes an outer membrane protein exposed on the bacterial cell surface. AB - A gene designated as aopB was identified which was involved in tumorigenesis of Agrobacterium tumefaciens. aopB is located on the circular chromosome as a single copy. This gene shares high homology with ropB, a Rhizobium leguminosarum gene encoding an outer membrane protein. A transposon mutant CGI1 containing a gfp tagged transposon insertion at aopB caused attenuated tumors on plants when inoculated at a low cell concentration (5x10(7) cells/ml). The mutation did not affect the bacterial growth on different media. A broad host range plasmid containing the wild type aopB could restore the tumor formation ability of CGI1 to the wild type level. When both aopB-gfp and aopB-phoA fusions were used to study the aopB gene expression, we found that the aopB gene was inducible by acidic pH but not by plant phenolic compound acetosyringone. aopB encodes a putative protein of 218 amino acids with a predicted molecular weight of 22.8 kDa. TnphoA transposon mutagenesis of aopB, subcellular fractionation and whole cell ELISA experiments indicated that AopB is an outer membrane protein exposed on the bacterial cell surface. It appeared that AopB was exclusively present in the outer membrane and not in other fractions. The vir gene induction assays showed that the aopB gene was not required for the expression of the Ti plasmid encoded vir genes that are essential for tumorigenesis. The C-terminal half of AopB is slightly homologous to some of the bacterial porin proteins and some of plant dehydrins. The role of AopB in Agrobacterium-plant interaction is discussed. PMID- 11891053 TI - Universal Fast Walking for direct and versatile determination of flanking sequence. AB - We report a highly compact system for accelerating direct genome walking. Unlike previous walking techniques, our strategy does not rely on restriction enzymes or ligases, and is therefore unaffected by the availability of useful restriction sites in the flanking region. A complete circumvention of molecular cloning steps qualifies this method for sequencing genome segments that are regarded unclonable, and thus unsequenceable by the traditional methods. A premium was placed on economy of design: the system comprises just four direct reagent additions, in microliter-scale volumes, over the course of a 6-h procedure. The walk range in this method is directly related to the capabilities of the associated polymerase blend, indicating that it can achieve in excess of 35 kilobases per reaction. It also produces a DNA fingerprint that is distinctive to the flanking sequence. Despite the complexity of banding patterns in these fingerprints, we observed that the reaction products were directly sequenceable. In view of its speed, reliability and generality, we term the described method Universal Fast Walking. PMID- 11891054 TI - Genetic linkage analysis to identify a gene required for the addition of phosphoethanolamine to meningococcal lipopolysaccharide. AB - Lipopolysaccharide (LPS) is important for the virulence of Neisseria meningitidis, and is the target of immune responses. We took advantage of a monoclonal antibody (Mab B5) that recognises phosphoethanolamine (PEtn) attached to the inner core of meningococcal LPS to identify genes required for the addition of PEtn to LPS. Insertional mutants that lost Mab B5 reactivity were isolated and characterised, but failed to yield genes directly responsible for PEtn substitution. Subsequent genetic linkage analysis was used to define a region of DNA containing a single intact open reading frame which is sufficient to confer B5 reactivity to a B5 negative meningococcal isolate. The results provide an initial characterisation of the genetic basis of a key, immunodominant epitope of meningococcal LPS. PMID- 11891055 TI - Characterization and expression of the human gene encoding two translocation liposarcoma protein-associated serine-arginine (TASR) proteins. AB - Translocation liposarcoma protein (TLS)-associated serine-arginine (TASR)-1 and 2 are two newly identified serine-arginine splicing factors. Our recent studies suggest that disruption of TASR-mediated pre-mRNA splicing is involved in the pathogenesis of human leukemia and sarcomas. The mRNA transcripts for TASR-1 and 2 share an identical sequence at the 5' untranslated region (5' UTR) and in part of the coding region; however the other regions of the transcripts diverge from each other and it was not clear whether the differences resulted from alternative splicing or transcription from two distinct genes. Here we describe the assignment of both TASR cDNAs to the same 16 kb DNA segment located on chromosome 1. Despite the presence of at least three retroposed products of TASR-1 mRNA in the human genome, only the 16 kb structural TASR gene on chromosome 1 is actively transcribed. In addition, multiple polyadenylation sites and a rare U12-type intron were found within the TASR gene. Transcription initiation site of the TASR gene was determined by primer extension; analysis of the TASR promoter revealed that it lacks the TATA box but contains a GC-rich sequence. When cloned into a luciferase reporter and transfected into human cells, the TASR promoter construct generated luciferase activity that was at least 2000 fold greater than the promoterless plasmid. Northern blot analysis showed that at least five different TASR-1 and -2 transcripts are expressed in a broad range of human tissues. PMID- 11891056 TI - pido, a non-long terminal repeat retrotransposon of the chicken repeat 1 family from the genome of the Oriental blood fluke, Schistosoma japonicum. AB - A newly described non-long terminal repeat (non-LTR) retrotransposon element was isolated from the genome of the Oriental schistosome, Schistosoma japonicum. At least 1000 partial copies of the element, which was named pido, were dispersed throughout the genome of S. japonicum. As is usual with non-LTR retrotransposons, it is expected that many pido elements will be 5'-truncated. A consensus sequence of 3564 bp of the truncated pido element was assembled from several genomic fragments that contained pido-hybridizing sequences. The sequence encoded part of the first open reading frame (ORF), the entire second ORF and, at its 3' terminus, a tandemly repetitive, A-rich (TA(6)TA(5)TA(8)) tail. The ORF1 of pido encoded a nucleic acid binding protein and ORF2 encoded a retroviral-like polyprotein that included apurinic/apyrimidinic endonuclease (EN) and reverse transcriptase (RT) domains, in that order. Based on its sequence and structure, and phylogenetic analyses of both the RT and EN domains, pido belongs to the chicken repeat 1 (CR1)-like lineage of elements known from the chicken, turtle, puffer fish, mosquitoes and other taxa. pido shared equal similarity with CR1 from chicken, an uncharacterized retrotransposon from Caenorhabditis elegans and SR1 (a non-LTR retrotransposon) from the related blood fluke Schistosoma mansoni; the level of similarity between pido and SR1 indicated that these two schistosome retrotransposons were related but not orthologous. The findings indicate that schistosomes have been colonized by at least two discrete CR1-like elements. Whereas pido did not appear to have a tight target site specificity, at least one copy of pido has inserted into the 3'-untranslated region of a protein-encoding gene (GenBank AW736757) of as yet unknown identity. mRNA encoding the RT of pido was detected by reverse transcription-polymerase chain reaction in the egg, miracidium and adult developmental stages of S. japonicum, indicating that the RT domain was transcribed and suggesting that pido was replicating actively and mobile within the S. japonicum genome. PMID- 11891057 TI - Cloning of rat thymic stromal lymphopoietin receptor (TSLPR) and characterization of genomic structure of murine Tslpr gene. AB - Thymic stromal derived lymphopoietin receptor (TSLPR) is a novel receptor subunit that is related in sequence to the interleukin (IL)-2 receptor common gamma chain. TSLPR forms a heterodimeric complex with the IL-7 receptor alpha chain to form the receptor for thymic stromal derived lymphopoietin, a cytokine involved in B- and T-cell function. We have cloned the TSLP receptor from rat and find that the WSXWX motif commonly found in extracellular domains of cytokine receptors is conserved as a W(T/S)XV(T/A) motif among TSLP receptors from mouse, rat and human. As in the mouse, TSLP receptor is widely expressed in rats suggesting that TSLPR may have roles in signaling outside the hematopoietic system. A zooblot analysis revealed that TSLPR is expressed in all vertebrate species examined. The absence of TSLPR in Saccharomyces cerevisiae, Drosophila melanogaster and Caenorhabditis elegans genomes is similar to the expression of several other cytokine receptors that have been characterized thus far. We have also characterized the genomic structure of the murine Tslpr gene which shows that in addition to primary sequence homology, it shares a common genomic organization of coding exons with the murine IL-2 receptor common gamma chain (Il2rg). Use of an alternative splice acceptor site leads to two alternatively spliced transcript variants of murine TSLPR, both of which are functional receptors. Finally, using linkage analysis, we mapped the murine Tslpr gene to mouse chromosome 5 between the Ecm2 and Pxn genes. PMID- 11891058 TI - The human homologue of the mouse Surf5 gene encodes multiple alternatively spliced transcripts. AB - Hu-Surf5 is included within the Surfeit locus, a cluster of six genes originally identified in mouse. In the present study, we have cloned and characterized the Hu-Surf5 gene and its mRNA multiple transcripts. Comparison of the most abundant cDNA and genomic sequence shows that the Hu-Surf5 is spread over a region of approximately 7.5 kb and consists of five exons separated by four introns. The nucleotide sequence of the genomic region flanking the 3'-end of the Hu-Surf5 gene revealed the presence of a processed pseudogene of human ribosomal protein L21 followed by Hu-Surf6 gene. Only 110 bp separate the transcription start site of Hu-Surf5 and Hu-Surf3/L7a gene and the transcription direction is divergent. Earlier studies defined the 110 bp region essential for promoter activity of Hu Surf3/L7a. Here, we show that this region stimulates transcription with a slightly different efficiency in both directions. The bidirectional promoter lacks an identifiable TATA box and is characterized by a CpG island that extends through the first exon into the first intron of both genes. These features are characteristic of housekeeping genes and are consistent with the wide tissue distribution observed for Hu-Surf5 expression. Hu-Surf5 encodes three different transcripts, Surf-5a, Surf-5b, and Surf-5c, which result from alternative splicing. Two protein products, SURF-5A and SURF-5B have been characterized. Production of chimaeras between the full-length SURF-5A or SURF-5B and the green fluorescent protein (GFP) allowed to localize both proteins in the cytoplasm. PMID- 11891059 TI - Cloning and characterization of novel extensin-like cDNAs that are expressed during late somatic cell phase in the green alga Volvox carteri. AB - Asexual individuals of the green alga Volvox carteri consist of two cell types, somatic and reproductive cells. The somatic cells are terminally differentiated post-mitotic cells which undergo gradual senescence leading to cell death in every generation. To understand the gene expression programs associated with senescence of somatic cells, we cloned two cDNAs, LSG1 and LSG2, that are preferentially expressed during this late developmental stage. These two cDNAs were deduced to encode Pro-rich motifs characteristic of extensin proteins that are components of the extracellular matrix. LSG1 also resembled genes encoding plant pathogenesis-related protein 1 (PR-1), while LSG2 showed similarities with genes encoding matrix metalloproteinases, including a gamete lytic enzyme of Chlamydomonas. We also found that S9, one of the late somatic cDNAs previously cloned by Tam and Kirk (Dev. Biol. 145 (1991) 51), was deduced to encode a protein with a composition similar to LSG2. The expression of PR-1 and a matrix metalloproteinase-encoding gene has been shown to be induced during senescence in higher plants. These results indicate that some of the late somatic genes in V. carteri are related to the senescence-associated genes in higher plants. PMID- 11891060 TI - Structure of the human carboxypeptidase M gene. Identification of a proximal GC rich promoter and a unique distal promoter that consists of repetitive elements. AB - The human carboxypeptidase M (CPM) gene was found to encompass about 112.6 kb of genomic sequence, containing 11 exons of which eight (exons 2-9) are common to all transcripts and contain the entire coding region. We have cloned several alternative variants of CPM transcripts that result from differential promoter usage and alternative splicing. Although CPM belongs to the same metallocarboxypeptidase subfamily as CPE, their intron/exon structures differ significantly. Multiple transcription start sites were found in the CPM gene that cluster in two regions separated by about 30 kb and are flanked by two unique functional promoters. One ('proximal') is immediately upstream of the coding region and contains GC-rich sequences and a typical TATA box whereas the other ('distal') consists almost entirely of repetitive elements. Luciferase reporter assays with constructs of the promoter regions showed they were both quite active in several cell lines. However, the proximal promoter was much stronger than the distal one in two of the human cell lines tested (HepG2 and HEK293) whereas both promoters were highly and equally active in the human monocytic cell line THP-1, which has high constitutive expression of CPM. PMID- 11891061 TI - Identification of six novel genes by experimental validation of GeneMachine predicted genes. AB - In silico gene identification from finished and unfinished human genome sequence has become critically important in many projects seeking to gain insights into the gene content of genomic regions implicated in diseases. To establish limitations and criteria for in silico gene identification, and to identify novel genes of potential relevance to human prostate cancer and melanoma, 3 Mb of chromosome 1 sequence have been analyzed using GeneMachine. This program is a software suite comprising of sequence similarity programs and four gene identification programs. A total of 49 potential transcripts were selected and 37 of them were selected for experimental validation. We verified 16 of the predicted genes by experimental analysis. The comparison of the predicted transcripts with their cloned forms helped to refine predicted gene models as well as to identify splice variants for several of them. Although sequences matching with ten of our verified genes have been recently deposited in the GenBank, six of them remain novel. Our studies support the feasibility of identifying novel genes from regions of interest using draft human genome sequence. PMID- 11891062 TI - Studies of three genes encoding Cinnamomin (a type II RIP) isolated from the seeds of camphor tree and their expression patterns. AB - Cinnamomin, which has three isoforms, is a type II ribosome-inactivating protein (RIP) purified from the mature seeds of camphor tree (Cinnamomum camphora). In a previous study, an incomplete cDNA that encoded the A- and B-chain of Cinnamomin but lacked signal peptide sequence was cloned. In the present paper, its full length cDNA was obtained by 5' rapid amplification of cDNA ends (5'RACE). Subsequently, polymerase chain reaction (PCR) amplification of its genomic DNA was performed. Unexpectedly, sequence analysis of the PCR products revealed three cinnamomin genes with >98.0% sequence identity. One of them corresponded to the published cDNA and was designated as cinnamomin I, whereas the other two genes were named as cinnamomin II and cinnamomin III, respectively. RT-PCR amplification of the cDNAs of cinnamomin II and III manifested that these two genes were functional. The three genes have no intron. Three Cinnamomin precursors that were inferred from the cDNA sequence of three cinnamomin genes exhibited relatively high sequence homology with other type II RIPs. Northern blot analysis demonstrated that the cinnamomin genes only expressed in cotyledons of C. camphora seeds and the acmes of expression emerged at 75-90 DAF when seeds were close to maturity. It is proposed that the three cinnamomin genes may encode three isoforms of Cinnamomin. The physiological function of Cinnamomin in C. camphora seeds is briefly discussed. PMID- 11891063 TI - Molecular characterization of a novel protein disulfide isomerase in carrot. AB - A protein disulfide isomerase (PDI) coding sequence was cloned from a cDNA library derived from carrot (Daucus carota L.) somatic embryos. The cDNA is 2060 bp in length and encodes for a protein of 581 amino acids and molecular weight of 64.4 kDa. Primary structure analysis of the deduced protein revealed two thioredoxin-like active sites and an endoplasmic reticulum-retention signal at its C-terminus, which is also found in PDIs in plants and animals. Although between the carrot protein and other plant PDIs there is only about 30% identity, the active site regions are almost identical. The corresponding mRNA was found in varying amounts, in all tissues investigated. A recombinant protein expressed from the carrot cDNA clone effectively catalyzed both glutathione-insulin transhydrogenation and the oxidative renaturation of denatured RNase A. These results suggest that the protein coded for by the carrot gene is a novel member of the PDI family in plants. We therefore designated this novel carrot gene PDIL1. The protein expressed by the PDIL1 cDNA sequence had a highly acidic stretch at its N-terminal region (no such domain exists in known plant PDIs), and was located far from known plant PDIs on a maximum likelihood tree. The PDIL1 gene, together with closely-related genes identified in Arabidopsis and tomato, was suggested to belong to a novel subfamily of PDIs. PMID- 11891064 TI - Transformation of Paramecium caudatum with a novel expression vector harboring codon-optimized GFP gene. AB - We have developed a novel expression vector, pTub-tel3, for transformation in Paramecium caudatum. The vector was constructed by cloning P. caudatum alpha tubulin 5' and 3' non-coding regions. To examine transformation with the pTub tel3 construct, we chose the green fluorescent protein (GFP) as a selection marker. When a linearized pTub-tel3 vector containing a GFP open reading frame was injected into the macronucleus, the GFP transcript was expressed in many clones whereas protein expression was detected only after extensive optimization of original GFP codons. GFP-derived fluorescence was distributed throughout the nuclei and cytoplasm except for contractile and food vacuoles. Upon continuous cell division, notable heterogeneity of GFP fluorescence among descendants from the same transformant has emerged. This expression vector can be applied to the analysis of protein trafficking and localization in addition to exogenous gene expression in P. caudatum. PMID- 11891065 TI - Structural characterization of the mouse Girk genes. AB - Cardiac and neuronal G protein-gated potassium (K(G)) channels are activated by neurotransmitters such as acetylcholine, opioids, and dopamine. K(G) channel activation leads to an inhibition of synaptic transmission. K(G) channels are tetrameric complexes formed by assembly of G protein-gated, inwardly-rectifying potassium (K(+)) channel (GIRK) subunits. Four mammalian GIRK subunits (GIRK1-4) have been identified. In this study, we identify key features of the four mouse Girk genes including sequence, intron/exon structures, alternative splicing events, and candidate transcriptional start points. The mouse Girk genes are organized similarly, each containing four to seven exons. While the mouse Girk1 and Girk2 genes are relatively large (>100 kb), mouse Girk3 and Girk4 genes are compact (<20 kb). Multiple mRNA variants of Girk1, Girk3, and Girk4 were identified, existing by virtue of alternative splicing and/or usage of distinct transcription initiation sites. These findings should facilitate future studies aimed at understanding the transcriptional regulation of K(G) channels and their potential involvement in disease. PMID- 11891066 TI - Drug delivery to the nail following topical application. AB - The absorption of drugs into the nail unit, following topical application to the nail plate, is highly desirable to treat nail disorders, such as onychomycosis (fungal infections of the nail). Nail permeability is however quite low and limits topical therapy to early/mild disease states. In this paper, the recent research into ungual drug delivery is reviewed. The nail unit and the two most common diseases affecting the nail--onychomycosis and nail psoriasis--are briefly described to set the scene and to give an overview of the nature and scope of the problem. The factors, which affect drug uptake and permeation through the nail plate such as solute molecular size, hydrophilicity/hydrophobicity, charge, and the nature of the vehicle, are then discussed, followed by ways of enhancing drug transport into and through the nail plate. Finally, drug-containing nail lacquers which, like cosmetic varnish, are brushed onto the nail plates to form a film, and from which drug is released and penetrates into the nail, are reviewed. PMID- 11891067 TI - Evaluation of Pulsincap to provide regional delivery of dofetilide to the human GI tract. AB - Pulsincap formulations designed to deliver a dose of drug following a 5-h delay were prepared to evaluate the capability of the formulation to deliver dofetilide to the lower gastrointestinal (GI) tract. By the expected 5-h release time, the preparations were well dispersed throughout the GI tract, from stomach to colon. Plasma analysis permitted drug absorption to be determined as a function of GI tract site of release. Dofetilide is a well-absorbed drug, but showed a reduction in observed bioavailability when delivered from the Pulsincap formulations, particularly at more distal GI tract sites. Dispersion of the drug from the soluble excipient used in this prototype formulation relies on a passive diffusion mechanism and the relevance of this factor to the reduced extent and consistency of absorption from the colon is discussed. In these studies the effects of the degree of dispersion versus the site of dispersion could not be ascertained; nevertheless the scintigraphic analysis demonstrated good in vitro in vivo correlation for time of release from Pulsincap preparations. The combination of scintigraphic and pharmacokinetic analysis permits identification of the site of drug release from the dosage form and pharmacokinetic parameters to be studied in man in a non-invasive manner. PMID- 11891068 TI - The effect of ethanol on the simultaneous transport and metabolism of methyl p hydroxybenzoate in excised skin of Yucatan micropig. AB - The effects of ethanol on the simultaneous transport and metabolism of methyl p hydroxybenzoate (HBM) were investigated in the skin of Yucatan micropig in vitro. It was found that transesterification occurred in the permeation studies involving ethanol. This was confirmed by monitoring the flux of ethyl p hydroxybenzoate (HBE) into the receptor phase, as well as by monitoring the fluxes of HBM and p-hydroxybenzoic acid (HBA). The apparent flux of total HBM was decreased. The solubility of HBM increased with ethanol concentration, thus, the activity of HBM in ethanol solution became low because we used 10 mM HBM solution for permeation studies. The enhancement factor (E) was calculated to correct the activity. E increased with increasing the flux of ethanol, thus, ethanol may function as an enhancer of HBM transport. The hydrolysis of HBM to HBA was inhibited, whereas transesterification of HBM to HBE was induced at all concentrations of ethanol used (10-40%). The formation of HBE occurred much more readily than that of HBA at all concentrations of ethanol used. PMID- 11891069 TI - Studies on the development of oral colon targeted drug delivery systems for metronidazole in the treatment of amoebiasis. AB - The aim of the present study is to develop colon targeted drug delivery systems for metronidazole using guar gum as a carrier. Matrix, multilayer and compression coated tablets of metronidazole containing various proportions of guar gum were prepared. All the formulations were evaluated for the hardness, drug content uniformity, and were subjected to in vitro drug release studies. The amount of metronidazole released from tablets at different time intervals was estimated by high performance liquid chromatography method. Matrix tablets and multilayer tablets of metronidazole released 43-52% and 25-44% of the metronidazole, respectively, in the physiological environment of stomach and small intestine depending on the proportion of guar gum used in the formulation. Both the formulations failed to control the drug release within 5 h of the dissolution study in the physiological environment of stomach and small intestine. The compression coated formulations released less than 1% of metronidazole in the physiological environment of stomach and small intestine. When the dissolution study was continued in simulated colonic fluids, the compression coated tablet with 275 mg of guar gum coat released another 61% of metronidazole after degradation by colonic bacteria at the end of 24 h of the dissolution study. The compression coated tablets with 350 and 435 mg of guar gum coat released about 45 and 20% of metronidazole, respectively, in simulated colonic fluids indicating the susceptibility of the guar gum formulations to the rat caecal contents. The results of the study show that compression coated metronidazole tablets with either 275 or 350 mg of guar gum coat is most likely to provide targeting of metronidazole for local action in the colon owing to its minimal release of the drug in the first 5 h. The metronidazole compression coated tablets showed no change either in physical appearance, drug content or in dissolution pattern after storage at 40 degrees C/75% RH for 6 months. PMID- 11891070 TI - Effect of vehicles and penetration enhancers on the in vitro percutaneous absorption of tenoxicam through hairless mouse skin. AB - The effects of vehicles and penetration enhancers on the in vitro permeation of tenoxicam from saturated solutions through dorsal hairless mouse skin were investigated. Various types of vehicles, including ester-, alcohol-, and ether types and their mixtures, were used as vehicles, and then a series of fatty acids and amines were employed as enhancers, respectively. Even though the fluxes of tenoxicam from saturated pure vehicles were generally low (0.1-1.1 microg/cm2 per h), the skin permeability of tenoxicam was significantly increased by the combination of diethylene glycol monoethyl ether (DGME) and propylene glycol monolaurate (PGML) or propylene glycol monocaprylate (PGMC); the highest fluxes were achieved at 40% of DGME in both of the two cosolvents. The marked synergistic enhancement was also obtained by using propylene glycol (PG)-oleyl alcohol (OAl) cosolvent. The greatest flux was attained by the addition of unsaturated fatty acids at 3% concentration to PG. But saturated fatty acids failed to show a significant enhancing effect. The enhancement factors with the addition of oleic acid (OA) or linoleic acid (LOA) to PG were 348 and 238, respectively. Tromethamine (TM) showed an enhancing effect by the increased solubility; however, triethanolamine (TEA) did not show a significant enhancing effect. Rather, it decreased the fluxes of tenoxicam when added to PG with fatty acids. The above results indicate that the combinations of lipophilic vehicles like OA, LOA or OAl and hydrophilic vehicles like PG can be used for enhancing the skin permeation of tenoxicam. PMID- 11891071 TI - The inhibition of phagocytosis of respirable microspheres by alveolar and peritoneal macrophages. AB - Respirable poly(lactic co-glycolic acid) (PLGA) microspheres (2-3 microm diameter), were fabricated as a model drug delivery system whose uptake by macrophages could be quantified by fluorescent activated cell sorting. The microspheres exhibited minimal release of the entrapped flourophore (rhodamine B) and thus avoided possible fluid phase uptake of the flourophore. Externally bound microspheres were removed from the cell membrane by acid washing. The fluorescent intensity associated with the cells arose, therefore, from the internalised microspheres. NR8383 continuous culture alveolar macrophages were verified against primary cultures as a good model of alveolar phagocytosis. Peritoneal macrophages were also isolated and systemic and alveolar phagocytosis compared. Poloxamer 338 adsorbed at the microsphere surface did not reduce phagocytosis by NR8383 macrophages. It did, however, reduce the number of microspheres contained in primary alveolar macrophages but did not reduce the percentage of phagocytic cells. Poloxamer coatings did not reduce phagocytosis by peritoneal macrophages once the ratio of five microspheres per cell was exceeded. Dipalmitoylphosphatidylcholine (DPPC), the major component of lung surfactant, was added to cultures to model the alveolar environment where it was observed to reduce phagocytosis. In light of this finding, microspheres were coated in DPPC, which reduced their uptake by all cell types at all microsphere to cell ratios. PMID- 11891072 TI - Effect of additives on the crystallization and the permeation of ketoprofen from adhesive matrix. AB - The crystallization of drug in a matrix may significantly affect the efficacy and quality of the transdermal drug delivery system. Therefore, the control of drug crystallization is of particular interest in the development of efficient transdermal delivery systems. In this study, we investigated the effects of various additives on the crystallization of ketoprofen in polyisobutylene (PIB) adhesive matrix. The effects of various additives on the permeation of ketoprofen from PIB matrix across hairless mouse skin were also examined. Poly(vinyl pyrrolidone) (PVP) K-30 was found to be the most effective crystallization inhibitor. Also, Poloxamer, Tween 80 and Labrasol significantly inhibited the crystallization of ketoprofen in a PIB matrix. In case of Tween 80, Labrasol, and PVP K-30, the flux of ketoprofen decreased as the loading content of the additives increased. However, the addition of Tween 80, Labrasol, or PVP K-30 significantly reduced the decrease in the flux of ketoprofen within the PIB matrix during a storage time of 3 weeks. PMID- 11891073 TI - An in vitro model for investigating the gastric mucosal retention of 14C-labelled poly(acrylic acid) dispersions. AB - Polymers that bind from solution onto gastric mucosae can be used as a means of facilitating localised drug delivery, or act as therapeutic agents in their own right (e.g. by forming a protective layer or by inhibiting enzymes). Previous workers have used semi-quantitative methods to identify the ability of commercially available poly(acrylic acid)s to bind to gastric mucosa. In this study, the binding and retention of labelled poly(acrylic acid)s to sections of gastric mucosa from the pyloric region of pigs stomach were evaluated using 'static' and 'dynamic flow' test systems. Dispersions (3%) of 'low', 'high' and 'ultra high' (cross-linked) polymers were seen to adhere to porcine pyloric mucosa after exposure and rinsing in the 'static' system. The high molecular weight polymer showed the greatest retention in the 'dynamic' test system when washing continuously with simulated gastric acid. Changing the pH of the dispersions from 4.3 to 6.2 had little effect on polymer retention. It was concluded that polymers that were sufficiently mobile in solution to spread on, and interact with, the mucosal surface, but had a sufficiently high molecular weight to form viscous solutions and/or bioadhere to the mucosa, may be retained on the mucosal surface for the longest periods. PMID- 11891074 TI - Altered immune response to liposomal allergens of Aspergillus fumigatus in mice. AB - Aspergillus fumigatus has been implicated as the major pathogenic fungus causing Aspergillus-mediated disorders. It secretes complex glycoprotein antigens and allergens, which induce type I and type III mediated hypersensitivity reactions. The immune response to these allergens/antigens in allergic disorders is characterized by elevated levels of specific IgE, Th2 cytokines and eosinophilia. In the current study, the ability of negatively charged liposomes entrapped with glycoprotein antigens and allergens of A. fumigatus to modulate the immune response was studied. Immune response in mice was evaluated with both free and liposomal formulations. Liposome entrapped glycoprotein antigens/allergens of A. fumigatus elicited a Th1 type response with increased levels of TNF-alpha (5.5 folds), IFN-gamma (four-folds), specific IgG (three-folds) and IgG2a (2.4-folds), low titers of specific IgG1 (2.2-folds decrease) and IgE (three-folds decrease), and decreased peripheral eosinophilia by four-folds in comparison to mice receiving free glycoprotein allergens/antigens of A. fumigatus. Histopathological examination of lung tissue sections clearly indicated reduced eosinophil infiltration in mice immunized with liposomal formulations. These results suggest potential of liposomal formulations for A. fumigatus allergens/antigens for exploration in immunotherapy. PMID- 11891075 TI - Solid-state characterization of nifedipine solid dispersions. AB - The purpose of this study is to characterize the nature and solid-state properties of a solid dispersion system of nifedipine (33.3% w/w) in a polymer matrix consisting of Pluronic F68 (33.3% w/w) and Gelucire 50/13 (33.3% w/w). The nature of nifedipine dispersed in the matrix was studied by powder X-ray diffractometry (PXRD), differential scanning calorimetry (DSC) and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS). The rate and extent of water uptake of the solid dispersion were determined by weight gain. The dissolution rate of nifedipine solid dispersion was determined using Apparatus 2 of USP XXIII (1995). Quantitative PXRD showed that the saturation solubility of nifedipine in the polymer matrix is 2.1-3.0% w/w and indicated an excess of crystalline nifedipine in the solid dispersion. The maximum water uptake by the solid dispersion exposed to 75% RH at 45 degrees C was 3.3 times higher than for the dispersion exposed to 65% RH at 25 degrees C. Over 8 weeks, PXRD and DRIFTS of the nifedipine matrix stored at 25 or 4 degrees C were unchanged, showing constancy of crystallinity and intermolecular interactions. For a given mass of nifedipine (20 mg) and for a given particle size of nifedipine (<850 microm), the initial release rate of nifedipine from the solid dispersion was faster (46.2% of the nifedipine dissolved in 20 min) than that of the pure drug (1.2% of the nifedipine dissolved in 20 min). The results indicate that the nifedipine solid dispersion is physically stable over 8 weeks. Nifedipine is released faster from the solid dispersion than from the pure crystalline drug of the same particle size. PMID- 11891076 TI - HEPC-based liposomes trigger cytokine release from peripheral blood cells: effects of liposomal size, dose and lipid composition. AB - The immune response caused by liposome stimulation was studied by assessing the level of several cytokines released from human peripheral blood cells. Liposome stimulation resulted in the release of IL-6, IL-10, IL-1beta, TNF-alpha and IFN gamma. The size of the liposomes affected the degree of the cytokine releases with larger sized liposomes causing higher levels of cytokine induction. In addition, it appears that the lipid composition of liposomes had no effect on the degree of cytokine release. The release of cytokines occurred even in the absence of serum, suggesting that serum proteins did not contribute to liposome stimulation in peripheral blood cells. The release of cytokines induced by liposome stimulation was inhibited by the presence of either protein kinase-C (PKC) or protein tyrosine kinase (PTK) inhibitor, but not by the presence of an endocytosis inhibitor. This indicates that signal transduction via PKC or PTK is necessary, in order for human peripheral blood cells to release cytokines (IL-6, IL-10, IL-1beta, TNF-alpha and IFN-gamma) as the result of liposome stimulation. These quantitative data on the release of cytokines by liposomal stimulation provide useful information for the development of rational drug delivery systems and the safety of cytokine induction via the use of liposomes. PMID- 11891077 TI - Dissolution testing of a poorly soluble compound using the flow-through cell dissolution apparatus. AB - Dissolution of Pfizer Compound PD198306, a poorly soluble compound, was studied in 25 mM pH 9 sodium phosphate solution with 0.5% SLS using the flow-through cell dissolution apparatus. Unmicronized and micronized drug powders were tested. Several methods of loading the drug powder into the flow-through dissolution cells and their impact on dissolution were investigated. The influence of flow rate of the dissolution medium on the rate and extent of dissolution were studied. PD198306 has poor wettability even in the presence of 0.5% SLS. It was found that loading the drug powder into the dissolution cell in the form of a suspension provided the best dissolution profile in terms of the rate and extent of dissolution. The flow rate of 4 ml/min resulted in good particle size discrimination. PMID- 11891078 TI - Determination of the optimal amount of water in liquid-fill masses for hard gelatin capsules by means of texture analysis and experimental design. AB - The aim of this study is to use texture analysis as a non-destructive test for hard gelatin capsules filled with liquid formulations to investigate mechanical changes upon storage. A suitable amount of water in the formulations is determined to obtain the best possible compatibility with the gelatin shell. This quantity of water to be added to a formulation is called the balanced amount of water (BAW). Texture profiling was conducted on capsules filled with hydrophilic polymer mixtures and with formulations based on amphiphilic masses with high HLB value. The first model mixture consisted of polyethylene glycol 400 and polyvinylpyrrolidone K17 with water and the second type consisted of caprylocaproyl macrogol glycerides (Labrasol) with colloidal silica (Aerosil 200) and water. The liquid-fill capsules were investigated by measuring changes on mass and stiffness after storage under confined conditions in aluminium foils. Capsule stiffness was investigated also as a parameter in a response surface analysis to identify the BAW. Polyvinylpyrrolidone did not show a great influence on the BAW in the range of 10-12% (w/w) for the first model mixture. Capsules with the less hydrophilic Labrasol formulations, however, kept their initial stiffness after storage best with only half of that amount, i.e. 5-6% (w/w) of water in the compositions. From this study it can be concluded that texture profiling in the framework of an experimental design helps to find hydrophilic or amphiphilic formulations that are compatible with gelatin capsules. Short-term stability tests are meaningful if capsule embrittlement or softening is due to water equilibration or another migration process that takes place rapidly. Long term stability tests will always be needed for a final statement of compatibility between a formulation and hard gelatin capsules. PMID- 11891080 TI - Antibacterial activity of Brazilian propolis and fractions against oral anaerobic bacteria. AB - Propolis collected from a cerrado area in Minas Gerais State, Brazil, was subjected to chromatography on silica gel column and to partition between immiscible solvents. Propolis aqueous-ethanolic extract and fractions obtained were tested for inhibitory activity against periodontitis-causing bacteria. All of the assayed bacterium species were susceptible to propolis extract. The two fractionation methodologies yielded fractions which were active against bacteria, with minimum inhibitory concentrations (MIC) ranging from 64 to 1024 microg/ml. TLC and HPLC analyses of the extract and of active fractions showed the presence of phenolic compounds of varied polarity. None of the assayed fractions was more active than the extract, suggesting that the antibacterial activity is probably due to the synergistic effect of several compounds. PMID- 11891081 TI - Assessment of reversible contraceptive efficacy of methanol extract of Mentha arvensis L. leaves in male albino mice. AB - The present study was undertaken to assess the reversible contraceptive efficacy of methanolic extract of Mentha arvensis leaves. Aqueous solution of the extract (10 mg per day per mouse) when administered orally to male mice of proven fertility for 20, 40 and 60 days caused inhibition of fertility while maintaining their normal sexual behaviour. With the increase in treatment duration, there occurred a corresponding decrease in the mean weight of testis and accessory organs of reproduction. Sperm concentration, motility and viability in the cauda epididymis were also decreased. Spermatozoa with coiled tails also appeared in the epididymal smear. However, all the induced effects returned to normalcy within 30 days following withdrawal of 60-day treatment. Oral administration of the extract also did not affect the body weight of the mice and their blood cells count, packed cell volume, haemoglobin and blood/serum biochemistry. PMID- 11891082 TI - Evaluation of immunomodulatory potential of Ocimum sanctum seed oil and its possible mechanism of action. AB - The present study investigates the effect of Ocimum sanctum seed oil (OSSO) on some immunological parameters in both non-stressed and stressed animals. An attempt has also been made to explore the possible mechanism of immunomodulatory activity. OSSO (3 ml/kg, ip) produced a significant increase in anti-sheep red blood cells (SRBC) antibody titre and a decrease in percentage histamine release from peritoneal mast cells of sensitized rats (humoral immune responses), and decrease in footpad thickness and percentage leucocyte migration inhibition (LMI) (cell-mediated immune responses). Restraint stress (RS) produced a significant reduction in the anti-SRBC antibody titre, foot pad thickness and percentage LMI (% LMI). The effects of RS on humoral as well as cell-mediated immune responses were effectively attenuated by pretreating the animals with OSSO. Co administration of diazepam (1 mg/kg, sc), a benzodiazepine (BZD), with OSSO (1 ml/kg, ip) enhanced the effect of OSSO on RS-induced changes in both humoral and cell-mediated immune responses. Further, flumazenil (5 mg/kg, ip), a central BZD receptor antagonist inhibited the immunomodulatory action of OSSO on RS-induced immune responsiveness. Thus, OSSO appears to modulate both humoral and cell mediated immune responsiveness and these immunomodulatory effects may be mediated by GABAergic pathways. PMID- 11891083 TI - Efficacy of some nupe medicinal plants against Salmonella typhi: an in vitro study. AB - Different medicinal plants: Euphobia hirta (Eh); Citrus aurantifolia (Ca), Cassia occidentalis (Co), and Cassia eucalyptus (Ce), which are claimed by the Nupes of Bida in Niger State of Nigeria to be effective in the treatment of typhoid fever were collected. Ethanolic and water extracts were obtained by standard procedures. Preliminary phytochemical screening showed that alkaloids were absent in all the parts of Eh, Co, and Ca studied, but present in all the parts of Ce studied. Saponins were absent in the leaves and florescence of Eh, Ca fruits, but present in little amounts in Eh and Co roots. Saponins were present in large amounts in all the parts of Ce studied. Tannins were in little amount in all the medicinal plants studied, except in Ca. Glycosides were not present in any of the medicinal plants studied. The in vitro and microbial analysis showed that only Ce showed inhibition to Salmonella typhi growth. Ca plus 'Kanwa' (a locally mined, alkaline salt) showed inhibition only at a high concentration of 'Kanwa'. The MIC and MBC of Ce are 1 and 2 mg/ml, respectively. The paper concludes that, of all the medicinal plants claimed by the Nupes to be effective against S. typhi, only Ce contains the natural compound that can be used in the treatment of typhoid fever. PMID- 11891084 TI - Screening of African medicinal plants for antimicrobial and enzyme inhibitory activity. AB - Seven plant species, belonging to different families, were collected in the eastern part of the Republic of Congo (Kivu) based on ethnopharmacological information. Their dichloromethane and methanolic extracts were tested for biological activity. Five of the seven collected plants exhibited antiplasmodial activity with IC(50) values ranging from 1.1 to 9.8 microg/ml. The methanolic extract of Cissampelos mucronata was the most active one showing activity against chloroquine sensitive (D6) and chloroquine resistant (W2) Plasmodium falciparum strains with IC(50) values of 1.5 and 1.1 microg/ml, respectively. Additionally, this extract significantly inhibited the enzyme tyrosine kinase p56(lck) (TK). The dichloromethane extract of Amorphophallus bequaertii inhibited the growth of Mycobacterium tuberculosis with a MIC of 100 microg/ml and the methanolic extract of Rubus rigidus inhibited the activity of both enzymes HIV1-reverse transcriptase (HIV1-RT) and TK p56(lck). PMID- 11891085 TI - Assessment of the antihypertensive and vasodilator effects of ethanolic extracts of some Colombian medicinal plants. AB - The antihypertensive and vasodilator effects of ethanolic extracts prepared from Calea glomerata Klatt, Croton schiedeanus Schlecht, Curatella americana L., Lippia alba (Mill)n N.E.Br. and Lupinus amandus, which are medicinal plants used in Colombian folk medicine for the treatment of hypertension, were assayed both in SHR and Wistar rats and in rat isolated aortic rings. At a dose of 20 mg/kg, intravenous bolus administration of the ethanolic extracts, from C. schiedeanus, C. americana and L. amandus showed significant antihypertensive activity in SHR, C. schiedeanus being the most active. C. schiedeanus elicited dose-dependent decreases in mean arterial pressure and heart rate (5-100 mg/kg, i.v.) in SHR but 200 mg/kg administered orally did not show any significant effects, even after 3 h of observation. In intact rat aortic rings, ethanolic extracts from C. schiedeanus and Calea glomerata relaxed the contractions induced by KCl (80 mM) and phenylephrine (10(-6) M) in a concentration-dependent manner (10(-6)-3x10(-4) g/ml), with IC(50) of 6.5x10(-5) (7.3-5.8) g/ml and 7.1x10(-5) (7.9-6.4) g/ml, respectively. Bioguided phytochemical fractionation of the ethanolic extract from C. schiedeanus was started. More than one active principle seems to be present, flavonoids and terpenoids compounds were detected. PMID- 11891086 TI - The effects of fruit essential oil of the Pimpinella anisum on acquisition and expression of morphine induced conditioned place preference in mice. AB - The problem of drug dependence still remains unresolved. In the present study the effects of an essential oil of Pimpinella anisum (Umbeliferae) on the expression and acquisition of conditioned place preference (CPP) induced by morphine in mice were investigated. Subcutaneous (s.c.) injections of morphine (2-5 mg/kg) produced place preference in a dose-dependent manner. Furthermore, intraperitoneal (i.p.) injection of the essential oil of P. anisum (0.125-0.5 ml/kg) induced conditioned place aversion (CPA). The mice which have received the essential oil of the P. anisum (0.125-0.5 ml/kg, i.p.) as well as the oil with morphine (5 mg/kg, s.c.) reduced the morphine effect. Administration of the essential oil of P. anisum (0.125-0.5 ml/kg, i.p.) on the test day did not show any effect on morphine action. It appeared that pre-administration with bicuculline (GABA(A) receptor antagonist) (1.5 mg/kg, i.p., 20 min before essential oil) diminished the effect of the essential oil of the P. anisum on morphine which induced CPP, but this result was not found for the GABA(B) receptor antagonist, CGP35348 (200 and 400 mg/kg, i.p., 10 min before essential oil). In conclusion, it appeared that the essential oil of the P. anisum may reduce the morphine effects via a GABAergic mechanism. PMID- 11891087 TI - Herbal medicines for sexually transmitted diseases and AIDS. AB - Sexually transmitted diseases (STDs) and acquired immunodeficiency syndrome (AIDS) are gaining significant importance at present due to rapid spread of the diseases, high cost of treatment, and the increased risk of transmission of other STDs and AIDS. Current therapies available for symptomatic treatment of STDs and AIDS are quite expensive beyond the reach of common man and are associated with emergence of drug resistance. Many patients of STDs and AIDS are seeking help from alternative systems of medicines such as Unani, Chinese, Ayurvedic, naturopathy, and homeopathy. Since a long time, medicinal plants have been used for the treatment of many infectious diseases without any scientific evidence. At present there is more emphasis on determining the scientific evidence and rationalization of the use of these preparations. Research is in progress to identify plants and their active principles possessing activity against sexually transmitted pathogens including human immunodeficiency virus (HIV) with an objective of providing an effective approach for prevention of transmission and treatment of these diseases. In the present review, plants reported to possess activity or used in traditional systems of medicine for prevention and treatment of STDs including AIDS, herbal formulations for vaginal application, and topical microbicides from herbal origin, have been discussed. PMID- 11891088 TI - Antiproliferative activity of the Netherlands propolis and its active principles in cancer cell lines. AB - The MeOH extract of the Netherlands propolis showed promising antiproliferative activity toward highly liver-metastatic murine colon 26-L5 carcinoma with an EC(50) value of 3.5 microg/ml. Further, antiproliferative activity-guided purification of the MeOH extract led us to isolate four flavonoids (1-4), seven cinnamic acid derivatives (5-11) and two new glycerol derivatives (12, 13), whose structures were elucidated on the basis of spectral analysis. The isolated compounds were tested for their antiproliferative activity against murine colon 26-L5, murine B16-BL6 melanoma, human HT-1080 fibrosarcoma and human lung A549 adenocarcinoma cell lines. The benzyl (9), phenethyl (10) and cinnamyl caffeates (11) possessed potent antiproliferative activities with EC(50) values of 0.288, 1.76 and 0.114 microM, respectively, toward colon 26-L5 carcinoma. These caffeates were considered to be active constituents of the Netherlands propolis in their antiproliferative activity. The antioxidative activity of these caffeates may play an important role in their antiproliferative activities. PMID- 11891089 TI - Ethnophysiology and herbal treatments of intestinal worms in Dominica, West Indies. AB - In rural Dominican ethnophysiology worms reside in a human organ called the 'worm bag'. Unchecked, worms can cause illness by growing in size and number, spreading out of the worm bag and into other organs. In this study of 'bush medicine', we use a measure of cognitive salience in free-listing tasks, which reveals five plants commonly used to treat intestinal worms. These were Ambrosia hispida (Asteraceae), Aristolochia trilobata (Aristlochiaceae), Chenopodium ambrosioides (Chenopodiaceae), Portulaca oleracea (Portulacaceae), and Artemisia absinthium (Asteraceae). Bioactive compounds appear to be present in all of these plants. The cognitive salience of these plant remedies coupled with evidence of biochemical properties suggest that they provide efficacious treatments for controlling intestinal parasite loads. PMID- 11891090 TI - Pharmacological effects of lavandulifolioside from Leonurus cardiaca. AB - Lavandulifolioside was detected for the first time in Leonurus cardiaca var. vulgaris [Moench] Briquet (Lamiaceae). The isolation was performed from the butanolic extract of the aerial parts and the identification by NMR and MS. The pharmacological properties of lavandulifolioside consist of significant negative chronotropism, prolongation of the P-Q, Q-T intervals and QRS complex, and decrease of blood pressure. Contrary to the butanolic extract lavandulifolioside did not reduce the spontaneous locomotor activity. In conclusion, the pharmacological pattern of lavandulifolioside did not explain the pharmacological effects of L. cardiaca L. alone. PMID- 11891091 TI - Isolation and identification of antibacterial compounds from Vernonia colorata leaves. AB - Vernonia colorata (Compositae) is used throughout Africa for a variety of ailments. This prompted the screening of this plant for biological activity. Previous experiments carried out in our laboratory revealed that the leaves possessed high antibacterial activity. Through conventional chromatographic techniques and bioassay-guided fractionation, the following sesquiterpene lactones were isolated and identified by spectroscopic data; vernolide (1), 11beta, 13-dihydrovernolide (2) and vernodalin (3). Only 2 is a novel compound, although its antibacterial activity is low compared to compounds 1 and 3 which had minimum inhibitory concentrations of 0.1-0.5 mg/ml against Gram-positive bacteria. PMID- 11891092 TI - Pharmacological and phytochemical screening of two Hyacinthaceae species: Scilla natalensis and Ledebouria ovatifolia. AB - Aqueous, ethanolic, dichloromethane and n-hexane extracts of Scilla natalensis Planch. and Ledebouria ovatifolia (Bak.) Jessop bulbs (Hyacinthaceae) were screened for antibacterial, anti-inflammatory, antischistosomic, anticancer and anthelmintic activity. Poor antibacterial activity against both Gram-positive and Gram-negative bacteria was shown with S. natalensis extracts. Good antibacterial activity was shown by the ethanolic and dichloromethane extracts of L. ovatifolia against Gram-positive bacteria. In the anti-inflammatory screening, the dichloromethane and hexane extracts of S. natalensis resulted in good inhibition against both COX-1 and COX-2. Ethanolic extracts had the highest inhibitory effect against nematodes in the anthelmintic assays. Poor anti-inflammatory and anthelmintic activity was found with L. ovatifolia. Aqueous extracts of S. natalensis had good activity against Schistosoma haematobium, with a minimum inhibitory concentration of 0.4 mg ml(-1). Aqueous extracts of fresh L. ovatifolia bulb material were found to be lethal to S. haematobium at a concentration of 1.6 mg ml(-1). The phytochemical screening of S. natalensis bulbs revealed the presence of saponins and bufadienolides within the bulbs. Bulbs of L. ovatifolia contained bufadienolides. PMID- 11891093 TI - Immunohistochemical study of muscle biopsy in children with cerebral palsy. AB - Muscle biopsy was examined in 20 children with cerebral palsy, using immunohistochemical methods for marker of denervation neural cell adhesion molecules (N-CAM) in addition to standard techniques. Histological and histochemical study showed mild myopathic changes, type 1 predominance, and type 1 and type 2 hypotrophy, in accord with previous observations. Immunohistochemical study showed N-CAM expression in most biopsies (15/20), usually in scattered fibers, whereas in four patients aged less than 6 years it was expressed in grouped fibers. Our study supports the hypothesis of motor unit remodeling as a consequence of spasticity, especially in early phases of the disease. PMID- 11891094 TI - Excessive expression of synaptojanin in brains with Down syndrome. AB - We investigated the expression of synaptojanin, which has been mapped on 21q22.2 on human chromosome, in the cerebral cortex of patients with Down syndrome (DS), using immunohistochemistry and immunoblotting. Synaptojanin expression was observed in Cajal-Retzius cells, cortical plate neurons, subplate neurons, intermediate neurons, germinal matrix cells and the ventricular neuroepithelium of the fetal cerebrum in both controls and DS. After birth, synaptojanin immunoreactivity was mainly observed in cytoplasm of cortical neurons and neurophils. These expressions of synaptojanin suggest a broader role in not only synaptic vesicle recycling, but also the regulation of neuronal migration and synaptogenesis in the fetal period. In comparison with controls, DS brains clearly showed higher immunoreactivity of synaptojanin in every structure, and most of the large neurons showed immunoreactivity. Western blotting with synaptojanin confirmed the increased expression in DS brains. Although the reason for excessive expression of synaptojanin in DS brains is obscured, one possibility can be explained on the basis of a gene dosage effect. As another possibility, on the assumption that synaptojanin modulates synaptic transmission and plays roles in clathrin-mediated synaptic vesicle endocytosis and signaling, the excessive expression of synaptojanin may be involved in compensatory mechanisms occurring in developing DS brains, such as neuronal loss, atrophic basilar dendrites, decreased spines and abnormal synaptic density and length. PMID- 11891095 TI - Sydenham's chorea: clinical findings and comparison of the efficacies of sodium valproate and carbamazepine regimens. AB - BACKGROUND: Sydenham's chorea is still the most frequently seen form of acquired chorea in childhood in developing world despite the use of antibiotics. It is a debilitating illness lasting for weeks or months and requires drug therapy. OBJECTIVE: To evaluate and compare the efficacies of sodium valproate and carbamazepine in the treatment of the choreiform movements in Sydenham's chorea. DESIGN: A prospective trial carried out with 24 children with Sydenham's chorea. PATIENTS: Twenty-four patients were divided into two groups having similar demographic and clinical properties. One group (n = 17) was given carbamazepine (15 mg/kg per day) and the other (n = 7) was given sodium valproate (20-25 mg/kg per day). As soon as the symptoms were taken under control, doses of the drugs were tapered slowly. The duration of the drug use was recorded. The time of response to therapy was compared between the groups and the patients were monitored for the adverse effects. RESULTS: There was no significant difference between the groups with respect to the time of clinical improvement and time of complete remission, duration of the therapy and the recurrence rates. Clinical improvement began by 8.0 +/- 4.0 days in sodium valproate and 7.4 +/- 8.2 days in carbamazepine group (P = 0.88). In the whole group no adverse effect was seen due to the drugs. CONCLUSION: Carbamazepine and valproic acid are equally effective and safe drugs in the treatment of choreiform movements in Sydenham chorea. PMID- 11891096 TI - The relationship between 99mTc-HMPAO brain SPECT and the scores of real life rating scale in autistic children. AB - Childhood autism is a developmental disability of unknown origin with probable multiple etiologies. The purpose of this study was to compare the changes of regional cerebral blood flow (rCBF) in autistic and non-autistic controls, and to determine the relationship between rCBF on 99mTc-hexamethylpropylene amine oxime (HMPAO) brain SPECT and the scores of the Ritvo-Freeman Real Life Rating Scale (RLRS), IQ levels, and age of autistic children. Eighteen autistic children (four girls, 14 boys; mean age: 6.13 +/- 1.99 years) and 11 non-autistic controls (five girls, six boys, mean age: 6.5 +/- 3.39 years) were examined using 99mTc-HMPAO brain SPECT. All the children satisfying DSM-IV criteria for autistic disorder were taken into evaluation, and scored by the Ritvo-Freeman RLRS. IQ levels of these children were determined by Goodenough IQ test. Six cortical regions of interest (ROIs; frontal (F), parietal (P), frontotemporal (FT), temporal (T), temporo-occipital (TO), and occipital (O)) were obtained on transaxial slices for count data acquisition. The ratio of average counts in each ROI to whole-slice counts for the autistic children was correlated with the scores of Ritvo-Freeman RLRS. Hypoperfusion in rCBF in autistic children compared with the control group were identified in bilateral F, FT, T, and TO regions. We found no relationship between rCBF on 99mTc-HMPAO brain SPECT and the scores of the Ritvo-Freeman RLRS. There was a relationship between bilateral F regions perfusion on 99mTc-HMPAO brain SPECT and the age of autistic children. There was also a negative correlation between IQ levels and the scores of sensory responses, social relationship to people, and sensory-motor responses. Our results suggest that 99mTc-HMPAO brain SPECT is helpful to locate the perfusion abnormalities but no correlation is found between rCBF on 99mTc-HMPAO brain SPECT and the scores of Ritvo-Freeman RLRS. PMID- 11891098 TI - Acute disseminated encephalomyelitis and poststreptococcal acute glomerulonephritis. AB - We experienced the case of a boy suffering from acute disseminated encephalomyelitis and concomitant acute glomerulonephritis. The multiple lesions observed on MR images, which located mainly in the cortical gray matter, quickly responded to methyl prednisolone pulse therapy. Renal biopsy confirmed the diagnosis of poststreptococcal acute glomerulonephritis. Streptococcus pyogenes was identified by pharyngeal culture, and the infection was serologically confirmed. We speculated that S. pyogenes infection was coincidentally involved in both diseases. PMID- 11891097 TI - The effects of antiepileptic drugs on spatial learning and hippocampal protein kinase C gamma in immature rats. AB - This study was conducted to determine if alterations in hippocampal protein kinase C (PKC) gamma is one of the cellular mechanisms by which conventional antiepileptic drugs affect learning and memory. Wistar Rats (21-day-old) were divided into five groups: (1) control (no training and drugs); (2) training group (no drugs); (3) phenobarbital (PB) group; (4) carbamazepine (CBZ) group; and (5) valproate (VPA) group. A hippocampus dependent learning task (spatial changing learning) was used in the latter four groups lasting a total of 10 days. Correct responding rate of training group was significantly higher (P < 0.05) than in the PB, CBZ and VPA group. The PKC gamma staining intensity in hippocampal CA1-2 region of training group was significant greater than that of the control and PB group. There was no difference in staining intensities between the CBZ, VPA group or training group. The amount of PKC gamma located in plasma membrane of hippocampal neurons was significantly higher in the training group (P < 0.05) than the control, PB and VPA groups. No differences were found between the training and CBZ group. Lastly, the amount of PKC gamma in cytosol of hippocampus did not significantly differ between any of the five groups. These results indicate that the three antiepileptic drugs used in this study all disturbed the spatial learning of immature rats. Spatial learning was concomitant with activation of PKC gamma in hippocampal neurons. PB and VPA likely adversely affect learning and memory by interfering with PKC gamma activation, whereas CBZ may act by a different mechanism, possibly in the post-translocation process or by a PKC gamma independent pathway. PMID- 11891099 TI - An autopsy case of ornithine transcarbamylase deficiency. AB - We present an autopsy case of ornithine transcarbamylase (OTC) deficiency with grumose degeneration in the dentate nucleus of the cerebellum. The patient had intractable neonatal convulsions and hyperammonemia from the 3rd day after birth. Diagnosis of OTC deficiency was made based on null activity of the enzyme and four-base deletions in exon 9 of the OTC gene. Death was due to sepsis as well as disseminated intravascular coagulation at 1 year and 2 months of age. Neuropathology showed multiple cystic changes and ulegyria in the bilateral frontal and parietal lobes. Multiple cysts were associated with the region, which was infiltrated with macrophages surrounded by astroglia showing palisading pattern. Ferrugination was marked in the thalamus and severe neuronal loss with astrogliotic change in the CA1-2 area of the hippocampus. Grumose degeneration was noted in the dentate nucleus of the cerebellum. This is the first report of grumose degeneration in OTC deficiency. PMID- 11891100 TI - Pharmacokinetics of zonisamide in perinatal period. AB - Zonisamide is widely used for intractable epilepsy and the effects of this drug on fetuses and neonates through the mother taking it for epilepsy need to be clarified. We measured the zonisamide concentration in plasma and breast milk using high-performance liquid chromatography to investigate the transfer of zonisamide through the placenta and breast milk, as well as its pharmacokinetics, in two neonates born to epileptic mothers receiving zonisamide. The transfer rates were 92% via the placenta and 41-57% through the breast milk. The first order kinetics of zonisamide in the two infants was elimination half-lives of 109 and 61h, respectively. PMID- 11891101 TI - Miller Fisher syndrome with transient coma: comparison with Bickerstaff brainstem encephalitis. AB - We herein report a 4-year-old boy with Miller Fisher syndrome (MFS) who presented with transient coma in addition to the typical triad of internal and external ophthalmoplegia, cerebellar ataxia and areflexia after an influenza type B infection. The electroencephalogram findings revealed intermittently generalized slow wave bursts. The cerebrospinal fluid revealed high protein and a lack of any cellular response. The serum anti-GQ1b IgG antibody was elevated in the acute phase and disappeared in the convalescent phase. The transient coma with the triad of MFS in this patient indicated an extended brainstem lesion including a reticular formation, which is also the responsible lesion of Bickerstaff brainstem encephalitis (BBE), but the magnetic resonance imaging repeatedly showed no abnormal finding. Our patient suggested the involvement of central nervous system in addition to the peripheral nerve injury in MFS. He also suggested that MFS and BBE may belong to the same group of disorders as syndrome of ophthalmoplegia, ataxia and areflexia (SOAA). PMID- 11891102 TI - Tubulo-interstitial nephritis caused by sodium valproate. AB - A severely handicapped 14-year-old Japanese girl had epilepsy and was treated with sodium valproate (SV) from the age of 7 years. Although the epileptic seizures were well controlled, she sometimes had a fever and hypokalemia from the age of 13 years. Laboratory examinations revealed metabolic acidosis, hypouricemia, hypophosphatemia, glycosuria, proteinuria and aminoaciduria, thus suggesting Fanconi syndrome. Gallium scanning showed marked renal uptake. A renal biopsy revealed interstitial nephritis without immuno-deposition. SV was replaced since it was considered to be the most probable cause of the renal involvement. Thereafter, she showed marked improvement of the clinical symptoms and the laboratory data gradually, and she never had a fever. Although SV is an effective anti-epileptic drug, we have to pay attention to adverse renal effects such as Fanconi syndrome and interstitial nephritis. PMID- 11891103 TI - Unilateral occlusion of the middle cerebral artery after varicella-zoster virus infection. AB - We report a 4-year-old child who developed hemiplegia 6 months after varicella zoster virus (VZV) infection. Cerebral angiography showed complete occlusion of the right middle cerebral artery with basal moyamoya vessels. Elevation of anti VZV antibody in the cerebrospinal fluid indicated central nervous system involvement. The association between VZV cerebral angitis and unilateral occlusion of right middle cerebral artery is discussed. PMID- 11891104 TI - Non-progressive viral myelitis in X-linked agammaglobulinemia. AB - We report a 14-year-old boy with X-linked agammaglobulinemia (XLA) complicated by isolated non-progressive myelitis caused by Coxsackie virus B1. Despite the absence of immunoglobulin supplement and persistence of the virus for the initial 2 years, motor impairment did not show any progression for 3 years. This report shows that the prognosis of central nervous system infection in XLA is not determined by immunoglobulin levels alone, and that it is not always progressive or fatal. The balance between host immunity and the virulence of the causative virus may be involved in the prognosis of meningoencephalitis in XLA. PMID- 11891105 TI - Acute disseminated encephalomyelitis associated with parainfluenza virus infection of childhood. AB - Acute disseminated encephalomyelitis associated with the parainfluenza virus has rarely been reported in childhood. A 2.5-year-old girl with acute disseminated encephalomyelitis, who developed bilateral symmetrical lesions in the basal ganglion, thalamus, corpus callosum, cerebral subcortical white matter, and cerebellar medulla on brain magnetic resonance imaging is described. Serological confirmation of parainfluenza virus infection was made 2 weeks following the onset of neurological symptoms. Four months later, the patient had a full recovery. At present, 3 years later, no relapse has been reported and she is leading a normal life. Our case is of interest because of its rarity, the striking brain magnetic resonance imaging, and the good neurological outcome. PMID- 11891106 TI - Henoch-Schonlein purpura with intracerebral hemorrhage. AB - We describe a case of Henoch-Schonlein purpura (HSP) with massive intracerebral hemorrhage (ICH) in a 7-year-old-girl. A cranial CT scan revealed extensive ICH in the left parietal region and right parieto-temporal through occipital regions. At the time of ICH onset, hypertension and coagulation abnormality were not observed, but factor XIII activity was markedly reduced 9%. ICH was thought to have resulted from a marked decrease in factor XIII. Factor XIII preparation was administered immediately after the onset of ICH, and enlargement of the hemorrhagic region was not seen. The present case is the only reported case of HSP complicated by ICH in which factor XIII level was measured. PMID- 11891107 TI - West syndrome and other infantile epileptic encephalopathies--Indian hospital experience. AB - Children with infantile epileptic encephalopathies comprising 3.5% of the Pediatric Neurology Clinic registrations in a tertiary care hospital were retrospectively analyzed. Data were retrieved from case records and analyzed for seizure semiology, prenatal and perinatal insults, developmental status and relevant investigations. The various therapeutic modalities and their influence on spasm frequency, long-term development and final seizure status were compared. The two primary outcome variables analyzed included final seizure status and developmental outcome. Of the 94 infantile epileptic encephalopathies, West syndrome was the commonest (55.3%), of which two thirds were symptomatic. Etiological factors were prenatal in 66.6% and perinatal in 33.3%. The initial response to ACTH was good in 54.5% with subsequent relapse in 27.8% and for prednisolone was 52.9 and 44.4%, respectively, compared to 25.3% spasms control with conventional antiepileptic drugs. Disease category of infantile epileptic encephalopathies evolved in 4, i.e. early myoclonic encephalopathy to West syndrome 1, early infantile epileptic encephalopathy to West syndrome 1, West syndrome to Lennox-Gastaut syndrome 2. Psychomotor retardation was seen in 88.2%, with 16.1% having normal development at onset of spasms. Microcephaly was associated with delayed development but did not influence final seizure outcome. Final seizure outcome was poor in children with delayed development at onset (adjusted odds ratio [OR] = 4), delay in diagnosis >12 months (OR = 2.27) and in children with Lennox-Gastaut syndrome (OR = 4.75). ACTH/prednisolone and antiepileptic treatment versus antiepileptics alone showed a good final seizure response in 36.6% versus 20%. Development on follow up was delayed in children with initial psychomotor retardation (OR = 23.4) and abnormal electroencephalogram (OR = 7.46). Perinatal factors constituted one third of symptomatic West syndrome. The use of ACTH/corticosteroids resulted in good initial spasm control though final seizure outcome and development were unaffected. Prednisolone had similar response to ACTH in spasm control but higher subsequent relapse rate. Vigabatrin was useful though often unaffordable. The identification of a neurometabolic etiology, though uncommon, has significant therapy implications. Delay in diagnosis was common and negatively influenced final seizure outcome. PMID- 11891111 TI - Self-perpetuating states in signal transduction: positive feedback, double negative feedback and bistability. AB - Cell signaling systems that contain positive-feedback loops or double-negative feedback loops can, in principle, convert graded inputs into switch-like, irreversible responses. Systems of this sort are termed "bistable". Recently, several groups have engineered artificial bistable systems into Escherichia coli and Saccharomyces cerevisiae, and have shown that the systems exhibit interesting and potentially useful properties. In addition, two naturally occurring signaling systems, the p42 mitogen-activated protein kinase and c-Jun amino-terminal kinase pathways in Xenopus oocytes, have been shown to exhibit bistable responses. Here we review the basic properties of bistable circuits, the requirements for construction of a satisfactory bistable switch, and the recent progress towards constructing and analysing bistable signaling systems. PMID- 11891112 TI - How signaling proteins integrate multiple inputs: a comparison of N-WASP and Cdk2. AB - Signal transduction proteins that can integrate multiple upstream signals play a critical role in the complex regulatory circuits that control cellular behavior. The two signaling node proteins cyclin-dependent kinase 2 and the actin regulator neuronal Wiskott-Aldrich syndrome protein have qualitatively similar signaling properties. Recent studies, however, reveal that these proteins utilize distinct mechanisms of signal integration, leading to subtle but important quantitative differences in behavior. PMID- 11891113 TI - The emerging power of chemical genetics. AB - Chemical genetic methods allow signal transduction pathways to be probed in a domain-specific manner. This subtle perturbation of function, when combined with classical genetic and biochemical data, allows for a better understanding of protein function. This in turn is leading to elucidation of pharmacological maps of signaling pathways. Recent studies have focused on diverse pathways, including the initiation of actin polymerization, oncogenic tyrosine kinase control of cell transformation, and molecular motor involvement in adaptation of sensory cells of the inner ear. PMID- 11891114 TI - Compartmentalisation of cAMP and Ca(2+) signals. AB - The available knowledge concerning second messengers such as Ca(2+) and cAMP has grown immensely in the past few years. The concept of tight spatial compartmentalisation of these signals within cells has led to more refined models of intracellular signalling. The development of recombinant probes based on the green fluorescent protein have allowed the monitoring of these second messenger levels in single cells, with high spatial and temporal resolution. PMID- 11891115 TI - Live-cell fluorescent biosensors for activated signaling proteins. AB - A new generation of live-cell fluorescent biosensors enables us to go beyond visualization of protein movements, to quantify the dynamics of many different protein activities. Alternate approaches can report post-translational modifications, ligand interactions and conformational changes, revealing how the location and subtle timing of protein activity controls cell behavior. PMID- 11891116 TI - "Omic" approaches for unraveling signaling networks. AB - Signaling pathways are crucial for cell differentiation and response to cellular environments. Recently, a large number of approaches for the global analysis of genes and proteins have been described. These have provided important new insights into the components of different pathways and the molecular and cellular responses of these pathways. This review covers genomic and proteomic (collectively referred to as "omic") approaches for the global analysis of cell signaling, including gene expression profiling and analysis, protein-protein interaction methods, protein microarrays, mass spectroscopy and gene-disruption and engineering approaches. PMID- 11891117 TI - Photosensory perception and signalling in plant cells: new paradigms? AB - Plants monitor informational light signals using three sensory photoreceptor families: the phototropins, cryptochromes and phytochromes. Recent advances suggest that the phytochromes act transcriptionally by targeting light signals directly to photoresponsive promoters through binding to a transcriptional regulator. By contrast, the cryptochromes appear to act post-translationally, by disrupting extant proteosome-mediated degradation of a key transcriptional activator through direct binding to a putative E3 ubiquitin ligase, thereby elevating levels of the activator and consequently of target gene expression. PMID- 11891118 TI - Structure of rhodopsin and the superfamily of seven-helical receptors: the same and not the same. AB - The crystal structure of rhodopsin provides significant insights concerning structure/activity relationships in visual pigments and related G-protein-coupled receptors. The specific arrangement of seven-transmembrane helices is stabilized by a series of intermolecular interactions that appear to be conserved among Family A receptors. However, the potential for structural and functional diversity among members of the superfamily of seven-helical receptors presents a significant future challenge. PMID- 11891120 TI - Roles of PI3Ks in leukocyte chemotaxis and phagocytosis. AB - Through selective disruption of phosphoinositide 3-kinase (PI3K) activity and the use of green fluorescent protein tagged derivatives of domains capable of specifically binding the lipid products of PI3Ks in vivo, it has been shown that this family of signalling enzymes have vital and distinct roles in chemotaxis, phagocytosis and phagosome maturation in leukocytes. PMID- 11891119 TI - Regulation of cell polarity during eukaryotic chemotaxis: the chemotactic compass. AB - Phosphatidylinositol 3-kinase lipid products and the Rho GTPases play a central role in transmitting information from chemotactic receptors to the effectors of cell polarity, and recent advances in the field have allowed us to understand these roles more clearly. Emergent properties of positive and negative regulation of these molecules may account for the establishment of cell polarity during chemotaxis for a wide range of cells from Dictyostelium to fibroblasts to neutrophils. PMID- 11891121 TI - Ion transport proteins anchor and regulate the cytoskeleton. AB - Structurally diverse ion transport proteins anchor the spectrin-actin cytoskeleton to the plasma membrane by binding directly to linker proteins of the ankyrin and protein 4.1 families. Cytoskeletal anchoring regulates cell shape and restricts the activity of ion transport proteins to specialised membrane domains. New directions are being forged by recent findings that localised anchoring by ion transport proteins regulates the ordered assembly of actin filaments and the actin-dependent processes of cell adhesion and motility. PMID- 11891122 TI - Axon guidance: the cytoplasmic tail. AB - Recent advances in the study of axon guidance have begun to clarify the intricate signalling mechanisms utilised by receptors that mediate path-finding. Many of these axon guidance receptors, including Plexin B, EphA, ephrin B and Robo, regulate the Rho family of GTPases, to effect changes in motility. Recent studies demonstrate a critical role for the cytoplasmic tails of guidance receptors in signalling and also reveal the potential for a great deal of crosstalk between the various receptor-signalling pathways. PMID- 11891123 TI - Receptor kinase signalling in plants and animals: distinct molecular systems with mechanistic similarities. AB - Plant genomes encode large numbers of receptor kinases that are structurally related to the tyrosine and serine/threonine families of receptor kinase found in animals. Here, we describe recent advances in the characterisation of several of these plant receptor kinases at the molecular level, including the identification of receptor complexes, small polypeptide ligands and cytosolic proteins involved in signal transduction and receptor downregulation. Phylogenetic analysis indicates that plant receptor kinases have evolved independently of the receptor kinase families found in animals. This hypothesis is supported by functional studies that have revealed differences between receptor kinase signalling in plants and animals, particularly concerning their interactions with cytosolic proteins. Despite these dissimilarities, however, plant and animal receptor kinases share many common features, such as their single membrane-pass structure, their inclusion in membrane-associated complexes, the involvement of dimerisation and trans autophosphorylation in receptor activation, and the existence of inhibitors and phosphatases that downregulate receptor activity. These points of convergence may represent features that are essential for a functional receptor kinase signalling system. PMID- 11891124 TI - A unified view of the DNA-damage checkpoint. AB - Recent investigation of the DNA-damage checkpoint in several organisms has highlighted the conservation of this pathway. The checkpoint's signal transduction pathway consists of four conserved classes of molecules: two large protein kinases having homology to phosphatidylinositol 3-kinases, three "sensor" proteins with homology to proliferating cell nuclear antigen, two serine/threonine (S/T) kinases, and two adaptors for the S/T kinases. This review compares the role of these four classes of checkpoint proteins in humans and model organisms. PMID- 11891125 TI - The mustard trypsin inhibitor 2 affects the fertility of Spodoptera littoralis larvae fed on transgenic plants. AB - The effects of mustard trypsin inhibitor MTI-2 expressed at different levels in transgenic tobacco lines have been evaluated by feeding the lepidopteran Spodoptera littoralis throughout its larval life. Specific conditions were selected to study the long-term effects of feeding larvae on transgenic plants expressing the inhibitor at various levels. The data obtained led to the establishment of three relevant parameters to be considered during the experimentation: (i) the PI content of the plant lines to be used; (ii) the developmental stage of larvae sensitive to that PI content; (iii) the ratio of MTI-2/proteases sufficient to inhibit gut proteases. The experimental data obtained from feeding S. littoralis larvae using these conditions led to two main results. First, when L2 S. littoralis larvae were fed on high MTI-2 expressing tobacco plants, no effects on larval development were detected but there was a significantly reduced fertility. When the same larvae were fed on low expressing MTI-2 tobacco plants, only a less marked lowering of fertility was observed. Second, after the first generation, no differences in protease activity were observed in insects derived from larvae fed on high or low MTI-2 expressing tobacco lines, suggesting that genetic traits observed in previous studies were not inherited. PMID- 11891126 TI - A coiled-coil region of an insect immune suppressor protein is involved in binding and uptake by hemocytes. AB - Polydnaviruses are associated with certain parasitoid wasps and are introduced into the body cavity of the host caterpillar during oviposition. Some of the viral genes are expressed in host tissues and corresponding proteins are secreted into the hemocoel causing suppression of the host immune system. The Cotesia rubecula polydnavirus gene product, CrV1, effectively inactivates hemocytes by mediating cytoskeleton breakdown. A precondition for the CrV1 function is the incorporation of the extracellular protein by hemocytes. Here, we show that a coiled-coil domain containing a putative leucine zipper is required for CrV1 function, since removal of this domain abolishes binding and uptake of the CrV1 protein by hemocytes. PMID- 11891127 TI - Protein kinase A: purification and characterization of the enzyme from two cold hardy goldenrod gall insects. AB - The catalytic subunit of protein kinase A (PKAc) was purified to apparent homogeneity from two species of cold-hardy goldenrod gall insects, Epiblema scudderiana and Eurosta solidaginis. Final specific activity for both enzymes was approximately 74.5 nmol of phosphate transferred per minute per milligram protein. Molecular weights were 41 and 40 kDa for E. scudderiana and E. solidaginis PKAc, respectively. K(m) values at 24 degrees C for the artificial substrate, Kemptide, were 38.1+/-4.9 and 3.67+/-0.11 microM for E. scudderiana and E. solidaginis PKAc, respectively, whereas K(m) Mg-ATP values were 61.1+/-6.9 and 30.7+/-4.1 microM. Assay at 4 degrees C lowered the K(m) for Kemptide of E. scudderiana PKAc by 55% and addition of 1M glycerol further lowered the K(m). Low assay temperature also enhanced holoenzyme dissociation in both species with the K(a) value for cyclic 3'5'-monophosphate at 4 degrees C lowered to just 13-18% of the value at 24 degrees C. Low temperature did not affect affinity for Mg-ATP or inhibition by PKA inhibitors (PKAi, H7, H89) but increased inhibition by some salts. PKAc from both species showed a break in the Arrhenius relationship at approximately 10 degrees C which suggests a conformational change at low temperature; activation energies (E(a)) were 2.2-3 fold higher for the lower (<10 degrees C) versus higher (>10 degrees C) range. Addition of naturally occurring polyols, 1M glycerol or 0.4M sorbitol, affected E(a) in some cases. Temperature dependent regulation of holoenzyme dissociation and PKAc kinetic properties may have an role in regulating the enzymes involved in polyol synthesis in cold-hardy insects. PMID- 11891128 TI - Aedes aegypti peritrophic matrix and its interaction with heme during blood digestion. AB - A large amount of heme is produced upon digestion of red cell hemoglobin in the midgut of mosquitoes. The interaction between heme and the peritrophic matrix (PM) was studied in Aedes aegypti. By light microscopy, the PM appeared as a light brownish layer between the intestinal epithelium and the alimentary bolus. This natural color can be attributed to the presence of heme bound to the matrix. In histochemical studies, a diffuse peroxidase activity of the heme molecules was clearly observed between the erythrocytes and the PM at 14 h after the blood meal. This activity tends to increase and concentrate in the PM reaching its maximum thickness at 24 h after feeding. Most of the heme of the PM was found associated to with enormous number of small electron-dense granules. The amount of heme bound to the PM increased in parallel with the progression of digestion, reaching a maximum at 48 h after feeding, when 18 nmol of heme were found in an individual matrix. The association of heme with PM from insects fed with plasma is saturable, suggesting the existence of specific binding sites for hemin in the PM. Taken all together, our data indicate that the PM performs a central role in heme detoxification in this insect. PMID- 11891129 TI - cDNA sequences and mRNA levels of two hexamerin storage proteins PinSP1 and PinSP2 from the Indianmeal moth, Plodia interpunctella. AB - In insects, storage proteins or hexamerins accumulate apparently to serve as sources of amino acids during metamorphosis and reproduction. Two storage protein like cDNAs obtained from a cDNA library prepared from fourth instar larvae of the Indianmeal moth (Plodia interpunctella) were cloned and sequenced. The first clone, PinSP1, contained 2431 nucleotides with a 2295 nucleotide open reading frame (ORF) encoding a protein with 765 amino acid residues. The second cDNA, PinSP2, consisted of 2336 nucleotides with a 2250-nucleotide ORF encoding a protein with 750 amino acid residues. PinSP1 and PinSP2 shared 59% nucleotide sequence identity and 44% deduced amino acid sequence identity. A 17-amino acid signal peptide and a molecular mass of 90.4 kDa were predicted for the PinSP1 protein, whereas a 15-amino acid signal peptide and a mass of 88 kDa were predicted for PinSP2. Both proteins contained conserved insect larval storage protein signature sequence patterns and were 60-70% identical to other lepidopteran larval storage proteins. Expression of mRNA for both larval storage proteins was determined using the quantitative reverse transcription polymerase chain reaction method. Only very low levels were present in the second instar, but both mRNAs dramatically increased during the third instar, peaked in the fourth instar, decreased dramatically late in the same instar and pupal stages, and were undetectable during the adult stage. Males and females exhibited similar mRNA expression levels for both storage proteins during the pupal and adult stages. The results support the hypothesis that P. interpunctella, a species that does not feed after the larval stage, accumulates these two storage proteins as reserves during larval development for subsequent use in the pupal and adult stages. PMID- 11891130 TI - The role of eicosanoids on Rhodnius heme-binding protein (RHBP) endocytosis by Rhodnius prolixus ovaries. AB - The participation of eicosanoids and second messengers on the regulation of RHBP endocytosis by the ovaries was investigated, using [(125)I]RHBP in experiments in vivo and in vitro. Addition of PGE(2) (one of the products of the cyclooxygenase pathway) decreased in vitro the uptake of RHBP by 35%. The rate of RHBP endocytosis increased in the presence of indomethacin, a potent cyclooxigenase inhibitor, up to 50% in vitro and up to 55% in vivo, thus giving support to the role of cyclooxygenase derivatives on endocytosis regulation. The amount of PGE(2) secreted to the culture medium by the cells of Rhodnius prolixus ovaries was 1.1 ng/ovary following RHBP uptake assay. The amount of PGE(2) decreases approximately 25% in the presence of 5 microM indomethacin. Using a scanning electron microscope we have observed that neither the surface area nor the patencies of follicle cells were affected by treatment with indomethacin, thus suggesting that, its effect is elicited in the oocyte. Finally, we have identified two ovarian peptides that were dephosphorylated after the indomethacin treatment (18 and 25 kDa). Taken together these data show that local mediators such as eicosanoids act upon the oocytes controlling RHBP endocytosis, perhaps using the protein phosphorylation signal transduction pathway. PMID- 11891131 TI - Voltage-dependent calcium channels in the corpora allata of the adult male loreyi leafworm, Mythimna loreyi. AB - In the corpora allata (CA) of the adult male loreyi leafworm, Mythimna loreyi, juvenile hormone acid (JHA) biosynthesis and release show a dose dependence on extracellular Ca(2+) concentration. Maxima are obtained with Ca(2+) concentrations of 2-10 mM, and synthesis and release are significantly inhibited under a Ca(2+)-free condition. The Ca(2+)-free inhibition of JHA release can be reversed by returning the glands to medium at 5 mM Ca(2+). The cytosolic free Ca(2+) concentration ([Ca(2+)](i)), which was measured with fura-2, in individual CA cells also shows a dose dependence on extracellular Ca(2+) concentration, with significant [Ca(2+)](i) depression being observed in the absence of extracellular Ca(2+). High K(+) significantly increases the JHA release and causes a transient [Ca(2+)](i) increase within seconds in CA cells. High-K(+)-stimulated JHA release is partially inhibited by the benzothiazepine (BTZ)-, dihydropyridine (DHP)- and phenylalkylamine (PAA)-sensitive L-type voltage-dependent calcium channel (VDCC) antagonists diltiazem, nifedipine and verapamil, respectively; by the N- and P/Q type VDCC antagonist omega-conotoxin (omega-CgTx) MVIIC; and by the T-type VDCC antagonist amiloride. The N-type antagonist omega-CgTx GVIA is the most potent in inhibiting the high-K(+)-stimulated JHA release. No inhibitory effect is shown by the P-type antagonist omega-agatoxin TK (omega-Aga TK). The high-K(+)-induced transient [Ca(2+)](i) increase is largely inhibited by the L-type antagonists (diltiazem, nifedipine, verapamil), by the N- and P/Q-type antagonist omega-CgTx MVIIC and by the T-type antagonist amiloride, and is totally inhibited by the N type antagonist omega-CgTx GVIA. No inhibitory effect is shown by the P-type antagonist omega-Aga TK. We hypothesize that L-type, N-type and T-type VDCCs may be involved to different degrees in the high-K(+)-stimulated JHA release and transient [Ca(2+)](i) increase in the individual CA cells of the adult male M. loreyi, and that the N-type VDCCs may play important roles in these cellular events. PMID- 11891132 TI - Isolation of a protein lethal to the endoparasitoid Cotesia kariyai from entomopoxvirus-infected larvae of Mythimna separata. AB - Virion-free plasma from entomopoxvirus (MyseEPV)-infected larvae of the armyworm, Mythimna separata, contains a factor that adversely affects the survival of the gregarious braconid endoparasitoid, Cotesia kariyai. Heating or proteinase K treatment eliminates the toxic effect of virion-free plasma on the parasitoid, suggesting that the lethal factor is a protein. We purified the Protein Lethal to C. kariyai larvae (PLCK) from the virion-free plasma of MyseEPV-infected M. separata larvae by a three-step procedure using gel filtration and cation exchange chromatography. Toxic activity was measured using an in vitro-cultured parasitoid bioassay. Parasitoid larvae cultured in IPL-41 medium (Weiss et al., In vitro 17 (1981), 495) containing 4.7 microg/ml purified PLCK shrank and died within 3days. The molecular weight of PLCK was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be about 28,000, under both reducing and non-reducing conditions, indicating that in its native form the protein is a single 28-kDa polypeptide. Western blot analysis indicated that the lethal protein is not present in the hemolymph of uninfected host larvae, but is induced in the hemolymph by infection with MyseEPV. Western blot analysis also indicated that the proteins of virions and occlusion bodies of MyseEPV are not serologically related to PLCK. PMID- 11891133 TI - Changes in protease activity and Cry3Aa toxin binding in the Colorado potato beetle: implications for insect resistance to Bacillus thuringiensis toxins. AB - Widespread commercial use of Bacillus thuringiensis Cry toxins to control pest insects has increased the likelihood for development of insect resistance to this entomopathogen. In this study, we investigated protease activity profiles and toxin-binding capacities in the midgut of a strain of Colorado potato beetle (CPB) that has developed resistance to the Cry3Aa toxin of B. thuringiensis subsp. tenebrionis. Histological examination revealed that the structural integrity of the midgut tissue in the toxin-resistant (R) insect was retained whereas the same tissue was devastated by toxin action in the susceptible (S) strain. Function-based activity profiling using zymographic gels showed specific proteolytic bands present in midgut extracts and brush border membrane vesicles (BBMV) of the R strain not apparent in the S strain. Aminopeptidase activity associated with insect midgut was higher in the R strain than in the S strain. Enzymatic processing of toxin did not differ in either strain and, apparently, is not a factor in resistance. BBMV from the R strain bound approximately 60% less toxin than BBMV from the S strain, whereas the kinetics of toxin saturation of BBMV was 30 times less in the R strain than in the S strain. However, homologous competition inhibition binding of (125)I-Cry3Aa to BBMV did not reveal any differences in binding affinity (K(d) approximately 0.1 microM) between the S and R strains. The results indicate that resistance by the CPB to the Cry3Aa toxin correlates with specific alterations in protease activity in the midgut as well as with decreased toxin binding. We believe that these features reflect adaptive responses that render the insect refractory to toxin action, making this insect an ideal model to study host innate responses and adaptive changes brought on by bacterial toxin interaction. PMID- 11891135 TI - Time-varying statistical dimension analysis with application to newborn scalp EEG seizure signals. AB - A new approach to the analysis of nonstationary possibly nonlinear time series is presented. It is based on an adaptive autocovariance eigenspectrum computation known as APEX together with the Rissanen's Minimum Description Length criterion for the selection of the most relevant eigenvalues. A new concept of time-varying instantaneous statistical dimension is introduced. The motivation for this new approach is the analysis of newborn electroencephalogram for which nonstationary is an inherent property. The proposed algorithm and new dimension are first assessed on synthetic data. Then, newborn scalp EEG data are analyzed using the proposed scheme. Transitions between different brain states are shown to occur on a baby having electrical and clinical seizures. PMID- 11891134 TI - Cloning and molecular characterization of two mosquito iron regulatory proteins. AB - Iron regulatory proteins (IRPs) control the synthesis of various proteins at the translational level by binding to iron responsive elements (IREs) in the mRNAs. Iron, infection, and stress can alter IRP/IRE binding activity. Insect messenger RNAs for ferritin and succinate dehydrogenase subunit b have IREs that are active translational control sites. We have cloned and sequenced cDNAs encoding proteins from the IRP1 family for the mosquitoes, Aedes aegypti and Anopheles gambiae. Both deduced amino acid sequences show substantial similarity to human IRP1 and Drosophila IRP1A and IRP1B, and all of the residues thought to be involved in aconitase activity and iron-sulfur cluster formation are conserved. Recombinant A. aegypti IRP1 binds to transcripts of the IREs of mosquito or human ferritin subunit mRNAs. No significant change in A. gambiae IRP1 messenger RNA could be detected during the various developmental stages of the life cycle, following iron loading by blood feeding, or after bacterial or parasitic infections. These data suggest that there is no change in gene transcription. Furthermore, bacterial challenge of A. gambiae cells did not change IRP1 protein levels. In contrast, IRP1 binding activity for the IRE was elevated following immune induction. These data show that changes in IRP1/IRE binding activity occur as part of the insect immune response. PMID- 11891136 TI - A discrete-time nonlinear Wiener model for the relaxation of soft biological tissues. AB - The present paper is devoted to introducing discrete-time models for the relaxation function of soft biological tissues. Discrete-time models are suitable for the analysis of sampled data and for digital simulations of continuous systems. Candidate models are searched for within both linear ARX structures and nonlinear Wiener models, consisting of an ARX element followed in cascade by a polynomial function. Both these discrete-time models correspond to sampling continuous-time exponential function series, thus preserving physical interpretation for the proposed relaxation model. The estimation data set consists of normalized stress relaxation curves drawn from experiments performed on samples of bovine pericardium. The normalized relaxation curves are found to be almost insensitive to both the magnitude of strain and the loading direction, and so a single model for the whole relaxation curves is assumed. In order to identify the parameters of the Wiener model an iterative algorithm is purposely designed. Over the ARX one, the nonlinear Wiener model exhibits higher capability of representing the experimental relaxation curves over the whole observation period. The stability of the solution for the iterative algorithm is assessed, and hence physical interpretation as material properties can be attached to the parameters of the nonlinear model. Suitable features of the Wiener model for computational application are also briefly presented. PMID- 11891137 TI - Short and long term non-linear analysis of RR variability series. AB - The complexity of RR variability is approached in the short and in the long term by means of black-box data analysis. Short term series of a few hundred beats are explored by means of informational entropy and predictability indexes. A correction to biases toward false determinism is performed assuming maximum uncertainty, whenever data do not furnish sufficient recurrences. Non-randomness and non-linearity are tested by means of surrogate data provided by random shuffling and phase randomization respectively. In the long term of the 24-h or of several hours, similar tests based on mutual information are applied and validated by means of surrogate series. In addition the state space reconstruction is carried out by means of state space non-linear filtering addressing directly the reconstructed trajectories. In this condition, parameters characterizing the hypothetical attractor, mainly the maximum Lyapunov exponent, can be reliably identified. PMID- 11891138 TI - Mutual information and phase dependencies: measures of reduced nonlinear cardiorespiratory interactions after myocardial infarction. AB - The heart rate variability (HRV) is related to several mechanisms of the complex autonomic functioning such as respiratory heart rate modulation and phase dependencies between heart beat cycles and breathing cycles. The underlying processes are basically nonlinear. In order to understand and quantitatively assess those physiological interactions an adequate coupling analysis is necessary. We hypothesized that nonlinear measures of HRV and cardiorespiratory interdependencies are superior to the standard HRV measures in classifying patients after acute myocardial infarction. We introduced mutual information measures which provide access to nonlinear interdependencies as counterpart to the classically linear correlation analysis. The nonlinear statistical autodependencies of HRV were quantified by auto mutual information, the respiratory heart rate modulation by cardiorespiratory cross mutual information, respectively. The phase interdependencies between heart beat cycles and breathing cycles were assessed basing on the histograms of the frequency ratios of the instantaneous heart beat and respiratory cycles. Furthermore, the relative duration of phase synchronized intervals was acquired. We investigated 39 patients after acute myocardial infarction versus 24 controls. The discrimination of these groups was improved by cardiorespiratory cross mutual information measures and phase interdependencies measures in comparison to the linear standard HRV measures. This result was statistically confirmed by means of logistic regression models of particular variable subsets and their receiver operating characteristics. PMID- 11891139 TI - Does synchronization reflect a true interaction in the cardiorespiratory system? AB - Cardiorespiratory synchronization, studied within the framework of phase synchronization, has recently raised interest as one of the interactions in the cardiorespiratory system. In this work, we present a quantitative approach to the analysis of this nonlinear phenomenon. Our primary aim is to determine whether synchronization between HR and respiration rate is a real phenomenon or a random one. First, we developed an algorithm, which detects epochs of synchronization automatically and objectively. The algorithm was applied to recordings of respiration and HR obtained from 13 normal subjects and 13 heart transplant patients. Surrogate data sets were constructed from the original recordings, specifically lacking the coupling between HR and respiration. The statistical properties of synchronization in the two data sets and in their surrogates were compared. Synchronization was observed in all groups: in normal subjects, in the heart transplant patients and in the surrogates. Interestingly, synchronization was less abundant in normal subjects than in the transplant patients, indicating that the unique physiological condition of the latter promote cardiorespiratory synchronization. The duration of synchronization epochs was significantly shorter in the surrogate data of both data sets, suggesting that at least some of the synchronization epochs are real. In view of those results, cardiorespiratory synchronization, although not a major feature of cardiorespiratory interaction, seems to be a real phenomenon rather than an artifact. PMID- 11891140 TI - Recurrence quantification analysis as a tool for nonlinear exploration of nonstationary cardiac signals. AB - The complexity, nonlinearity and nonstationarity of the cardiovascular system typically defy comprehensive and deterministic mathematical modeling, except from a statistical perspective. Living systems are governed by numerous, continuously changing, interacting variables in the presence of noise. Cardiovascular signals can be shown to be discontinuous alternations between deterministic trajectories and stochastic pauses (terminal dynamics). One promising approach for assessing such nondeterministic complexity is recurrence quantification analysis (RQA). As reviewed in this paper, strategies implementing quantification of recurrences have been successful in diagnosing changes in nonstationary cardiac signals not easily detected by traditional methods. It is concluded that recurrence quantification analysis is a powerful discriminatory tool which, when properly applied to cardiac signals, can provide objectivity regarding the degree of determinism characterizing the system, state changes, as well as degrees of complexity and/or randomness. PMID- 11891141 TI - Spontaneous and forced non-linear oscillations in heart period: role of the sino atrial node. AB - The aim of this study is to investigate whether and to which extent the non linearity of the sino-atrial node contributes to the complex interactions between autonomic control and heart period. A non-linear model of the autonomic control on the sino-atrial node has been analyzed under particular experimental conditions: the autonomic nervous system was forced to act at established frequencies, by a controlled breathing procedure and a non-invasive periodic modulation of carotid baroreceptors. We quantify the non-linear coupling between the stimuli and both the experimental and the simulated heart period fluctuations by using the recurrence plot quantification analysis. Our results show that the non-linear interaction between external stimuli and heart period fluctuations is due to a certain extent to the non-linear features of the sino-atrial node. PMID- 11891142 TI - Fractal dimensions of laser doppler flowmetry time series. AB - Laser Doppler flowmetry (LDF) provides a non-invasive method of assessing cutaneous perfusion. As the microvasculature under the probe is not defined the measured flux cannot be given absolute units, but the technique has nevertheless proved valuable for assessing relative changes in perfusion in response to physiological stress. LDF signals normally show pronounced temporal variability, both as a consequence of the pulsatile nature of blood flow and local changes in dynamic vasomotor activity. The aim of the present study was to investigate the use of methods of nonlinear analysis in characterizing temporal fluctuations in LDF signals. Data were collected under standardised conditions from the forearm of 16 normal subjects at rest, during exercise and on recovery. Surrogate data was then generated from the original time series by phase randomization. Dispersional analysis demonstrated that the LDF data was fractal with two distinct scaling regions, thus allowing the calculation of a fractal dimension which decreased significantly from 1.23 +/- 0.09 to 1.04 +/- 0.02 during exercise. By contrast, dispersional analysis of the surrogate data showed no scaling region. PMID- 11891143 TI - Improved estimators for fractional Brownian motion via the expectation maximization algorithm. AB - Fractional Brownian motion (FBM) provides a useful model for many physical and biological phenomena demonstrating long-term dependencies and 1/f-type spectral behavior. In this model, only one parameter is necessary to describe the complexity of the data, the Hurst exponent (H). The development of accurate estimators for H is a topic of interest in areas such as radiographic image processing and heart rate variability (HRV) analysis. The development of a 1D estimator utilizing the Expectation-Maximization (EM) algorithm is explained; the estimator is designed for a signal model consisting of FBM and additive white noise. The performance of this estimator is tested on simulated noisy FBM data sets, and found to provide more accurate estimates of H than a maximum likelihood estimator for FBM and the detrended fluctuation analysis (DFA) method. PMID- 11891144 TI - The 15-domain serine proteinase inhibitor LEKTI: biochemical properties, genomic organization, and pathophysiological role. AB - Proteinases are involved in specific and non-specific proteolytic reactions, and participate in many pathophysiological processes. Normally, they are regulated by endogenously produced proteinase inhibitors which, thus, represent lead structures for the development of therapeutics. We succeeded in partially isolating and cloning a novel human serine proteinase inhibitor which, according to its structure and the expression pattern of the corresponding gene, was termed lympho-epithelial Kazal-type-related inhibitor (LEKTI). This inhibitor is of special interest because it exhibits an extraordinarily large number of 15 potentially inhibitory domains and is of pathophysiological importance for the severe congenital disease Netherton syndrome. Here, we review the as yet known data on protein structure, biochemical properties, genomic organization and gene expression. Furthermore, the relevance of LEKTI for several disorders pointing out its possible future therapeutic value, is discussed. PMID- 11891145 TI - Dermatological diseases and signs of HIV infection. AB - Therapy of HIV infection has undergone significant changes since the introduction of highly-active antiretroviral therapy (HAART). Mortality and the appearance of opportunistic infections have significantly been reduced. Diseases of the skin and adjacent mucous membranes often provide the first signs for HIV infection. The spectrum of dermatologic findings related to HIV includes a variety of cutaneous and mucocutaneous disorders. The most frequent diagnoses are oral candidiasis, mollusca contagiosa, oral hairy leuokoplakia, herpes zoster and herpes simplex, seborrheic dermatitis, and Kaposi's sarcoma. Incompatibility reactions to drugs are observed on a strikingly frequent basis in HIV infection. Such severe incompatibility reactions are much more frequent in HIV patients than in the normal population. Inducers often include sulfonamides, cotrimoxazole, tuberculostatics as well as nucleoside-type reverse transcriptase inhibitors. PMID- 11891146 TI - Allergy, total IgE and eosinophils in East and West -- serious effects of different degrees of helminthiasis and smoking. AB - Allergies are rarer in East Germany than in West Germany, although elevated IgE levels and blood eosinophil counts can be found in the East. The aim of this study is to control the known results in Slowakia. The total-IgE and the blood eosinophil counts as well as stool samples of 311 plant-workers in Slovakia and 522 in West Germany were checked. In 1999/2000 21% of the plant workers examined in Slovakia suffered from an allergy compared with 32 % of the workers in West Germany. In contrast the total IgE in the East (Slovakia) was 166 IU/ml compared with 99 IU/ml in West Germany. Worms were detected in the stools in 7 % of those examined in the East and in only 1 % of those in the West. The percentage distribution of eosinophils in the blood was on average 7.5% in the East and 2.7% in the West. 39% of the workers in the East were smokers compared with 20% in the West. Nevertheless among non-smoking workers volunteers without parasites, the total IgE and eosinophil count were higher in those from the East than in those from the West. For Slowakia we can show for the first time that allergies are less frequent compared with West-Germany. The total IgE and the blood eosinophils are associated with higher smoking habits and more frequent helminthic diseases. PMID- 11891148 TI - Eradication of initial Pseudomonas aeruginosa colonization in patients with cystic fibrosis. AB - The optimal treatment for the eradication of initial P. aeruginosa infection in CF is still unclear. Recently long-term inhaled tobramycin has been proposed. Here we report the results with brief inhaled and/or systemic anti-pseudomonal treatments. Initial P. aeruginosa colonization was successfully eradicated as demonstrated by negative repetitive throat cultures or sputa and serum antipseudomonal antibodies in 15 of 17 patients for at least two years. Randomized, controlled trials are urgently needed to define the optimal protocol for the eradication of P. aeruginosa. Pseudomonas aeruginosa, infection, treatment, antibodies; inhaled tobramycin PMID- 11891147 TI - Inhalation of aerosolized vitamin a: reversibility of metaplasia and dysplasia of human respiratory epithelia -- a prospective pilot study. AB - The objective of this preliminary uncontrolled study was twofold: First, to assess the feasibility of retinyl palmitate inhalation and second, to analyze the changes of metaplastic lesions of the respiratory epithelium (metaplasia or dysplasia) following retinyl palmitate inhalation. The response to a daily dose of 18.000 I.U. retinyl palmitate by inhalation over a period of 3 month was assessed in 11 subjects (9 smokers, 2 ex-smokers). Using white-light bronchoscopy combined with autofluorescence bronchoscopy, bronchial biopsies were taken before and after a 3 month-period. The biopsy samples were evaluated blind by a referee lung pathologist. The overall response rate (remission or partial remission) was 56% (95% CI 0.30 0.79; p<0.05). These data suggest that inhalation of retinyl esters could be a promising therapeutical approach for chemoprevention of lung cancer. Vitamin A; chemoprevention; lung cancer; squamous metaplasia; dysplasia; retinoids PMID- 11891149 TI - Biocompatibility of dental materials in two human cell lines. AB - It was the aim of this study to investigate the biocompatibility of metallic (titan, gold, amalgam) and ceramic dental materials in contact with gingival (GF) and epithelial tumour cells (EpiCa). The cells were incubated with the test specimens (5 mg/piece) over a period of six days. Cellular proliferation rate, protein synthesis, and prostaglandin release (PGE2) served as parameters to determine the biocompatibility of the materials. - The investigations showed that the protein values were subject to slight variations following contact with the dental materials. Incubation of the cells with the test materials resulted in material dependent increases in PGE2-values (GF/EpiCa: titan: 187.4%, 131.0%; ceramic: 151.5%, 176.4%; gold: 114.5%, 123.8%; amalgam 150.6%, 159.8%). A comparison of control cells and cells of the test series (GF/EpiCa) after cell stimulation with 10-5M arachidonic acid (AA) showed the following changes in PGE2 on contact with titan (110.0%, 167.5%), ceramic (98.7%, 188.9%), gold (119.5%, 153.1%), and amalgam (68.9%, 179.5%). - Measurement of the proliferation rate (24 hours) further demonstrated that the dental materials used exerted an influence on the growth rates. While amalgam was associated with a marked reduction in the proliferation rate, titan, gold and ceramic induced only slight changes in the growth rate. - The results of the present study demonstrate that the cell culture systems used represent suitable in vitro models for the investigation of the biocompatibility of dental materials. The level of cell irritation was shown to be lowest for titan, closely followed by ceramic and gold. Cell culture; prostaglandin release; titan; ceramic; amalgam PMID- 11891150 TI - Substance use among Iranian cardiovascular patients. AB - AIMS: This study assessed the prevalence of substance use among Iranian patients with cardiovascular disease who were admitted in different cardiac wards at Shiraz general hospitals. DESIGN: Cross-sectional survey using structured interview and also using DSM-IV criteria for substance dependency. SETTING: General hospitals in Shiraz city (Patients with cardiovascular disease admitted in different cardiac wards). PARTICIPANTS: Ninety-six patients selected randomly. FINDINGS: Data were gathered by a structured interview from 96 patients admitted in cardiac ward of general hospitals in 2001. The mean age was 52.6 yr., (SD = 14.64) ranging from 15 to 100 yr. About 38.5% (58.3% of men and 18.8% of women) reported the use of substance(s) once or more in their lives. The majority (36.5%) used tobacco, 9.4% used opium and 8.3% used alcohol. No body used cocaine or LSD. About 24% (39.6% of men and 8.3% of women) were current substance dependent, using DSM-IV criteria. The majority (22%) were nicotine dependent, 5.2% were opium dependent and 3.1% were alcohol dependent. The reported reasons for initial use of substance(s), in order of frequencies, were Enjoyment, Modeling and Release of tension, and also for current users were, Habit, Enjoyment, Release of tension and Need. CONCLUSIONS: Substance use especially cigarettes, opium and alcohol was found to be high among patients. There was no report of cocaine or LSD use. Cultural attitudes toward substance use were found to affect the type and amount of use. These findings can be considered when planning preventive programs. cardiovascular disease; substance dependency; alcohol; tobacco; opium; heroin; LSD; cocaine; hashish; marijuana; morphine PMID- 11891151 TI - Assisting medical educators to foster cultural competence. PMID- 11891153 TI - Integrating social factors into cross-cultural medical education. AB - The field of cross-cultural medical education has blossomed in an environment of increasing diversity and increasing awareness of the effect of race and ethnicity on health outcomes. However, there is still no standardized approach to teaching doctors in training how best to care for diverse patient populations. As standards are developed, it is crucial to realize that medical educators cannot teach about culture in a vacuum. Caring for patients of diverse cultural backgrounds is inextricably linked to caring for patients of diverse social backgrounds. In this article, the authors discuss the importance of social issues in caring for patients of all cultures, and propose a practical, patient-based approach to social analysis covering four major domains--(1) social stress and support networks, (2) change in environment, (3) life control, and (4) literacy. By emphasizing and expanding the role of the social history in cross-cultural medical education, faculty can better train medical students, residents, and other health care providers to care for socioculturally diverse patient populations. PMID- 11891154 TI - Eradicating essentialism from cultural competency education. AB - An increasingly diverse society requires physicians to be able to competently treat those with whom they do not share ancestry and/or culture. Therefore, medical school educators need to train physicians who are capable of interacting appropriately and effectively with individuals from a broad array of populations and cultures. Such education cannot be simply a list of traits about other groups, as this may merely reinforce stereotypes. Instead, this education must expose and eradicate the existing essentialist biases in medicine. Essentialism, by focusing on differences, artificially simplifies individual and group identities and interactions. The essentialist viewpoint needs to be replaced with an ethnogenetic one, which recognizes that groups, cultures, and the individuals within them are fluid and complex in their identities and relationships. The ethnogenetic perspective must be fully integrated into medical education if medical schools are to produce physicians who will be truly qualified to give competent patient care in our increasingly complex societies. PMID- 11891155 TI - When is risk stratification by race or ethnicity justified in medical care? AB - Issues of race and ethnicity have been controversial in both clinical care and medical education. In daily practice, many physicians struggle to be culturally competent and avoid racial stereotyping. One educational development that makes this goal more complex is the rise of clinical epidemiology and Bayesian thinking. These population-based, probabilistic approaches to medicine help guide the diagnostic and therapeutic pathways for patients, and are foundations of the evidence-based medicine movement. Can Bayesian thinking be applied effectively to issues of race and ethnicity in medical care, or are the dangers of prejudicial stereotyping too great? The authors draw upon lessons from recent cases of racial profiling, and develop a conceptual framework for thinking about ethnicity as a clinical tool. In their typology of ethnicity as a proxy, they argue that the costs of using ethnicity as a proxy for socioeconomic status and behavior are too high, but that ethnicity may appropriately be used as an initial proxy for history, language, culture, and health beliefs. They discuss their approach within the context of new curricula in cultural competence, and argue that viewing the patient within a wider cultural setting can help guide the initial clinical approach, but individualized care is mandatory. Also, physicians must remain sensitive to the changing nature of cultural norms; thus lifelong learning and flexibility are necessary. PMID- 11891158 TI - Educating future physicians for a minority population: a French-language stream at the University of Ottawa. AB - The Faculty of Medicine at the University of Ottawa has recently developed a French-language undergraduate medical education stream in order to train physicians for the francophone minority population of the province of Ontario. This new program was planned with the following societal requirements in mind: the need to receive health care in one's mother tongue, the need to have physicians who know the community, and the expectation of receiving good medical care in an ambulatory setting. A systematic educational planning model was used in order to develop three educational innovations in response to these needs and expectations: a communication skills laboratory; early student exposure to the ambulatory, primary care setting for development of clinical skills; and clerkship rotations in a francophone community hospital. Program developers provided ongoing faculty development activities in order to prepare francophone faculty for their new roles. They also considered student participation in program development an essential element of its success. The program has positive outcomes both within and outside the Faculty of Medicine. These include an enrichment effect on the English-language stream, an increased interest in medical education, student satisfaction with their community hospital clerkship rotations, and the recognition of the educational program as a national resource for francophone minority groups. Medical schools that serve minority population groups may benefit from the experience gained at the University of Ottawa. PMID- 11891159 TI - Cultural sensitivity training in Canadian medical schools. AB - The authors describe the results of a survey they carried out in 2000 to determine the status of cultural sensitivity training in 16 Canadian medical schools using structured telephone interviews of associate and assistant deans or curriculum directors and curriculum coordinators. Their goal was to obtain a descriptive analysis of school-specific objectives, curriculum content, methods, and evaluation formats. The survey was prompted by the growing concern that in culturally diverse societies, medical education has failed to keep pace with the changing composition of the patient population. Only one of the eight schools that integrated cultural sensitivity within their objectives made explicit mention of the topic in its clerkship evaluation form. While seven of the 16 schools did not have any statement on cultural sensitivity in their curricular objectives, they integrated cultural sensitivity in their curricula using various educational methods, with PBL cases, lectures, and small-group discussions being the commonest formats. These educational methods were primarily offered to students in their first and second years. Student participation was required, but program lengths ranged from two to 40 hours. Additional findings for each school are presented. The authors conclude that while progress has been made, lack of adequate resources and a number of obstacles to inclusion of multicultural health content in curricula appear to remain ongoing problems. Further investment in faculty development and administrative staff support for a multicultural curriculum are needed, as is more research on effective curricular components. PMID- 11891157 TI - The hidden curriculum in multicultural medical education: the role of case examples. AB - Explicit cross-cultural learning experiences in medical education are provided within the context of implicit experiences provided by a greater "hidden curriculum." The authors conducted a content analysis of 983 cases presented in the 1996-1998 year one and year two curriculum at the University of Minnesota Medical School to determine in what ways they might embody elements of the hidden curriculum, i.e., how they either supported or undermined explicit messages about diverse patient populations. Cases were coded for demographic variables, potential risk factors, and diagnoses or presenting problems. The findings revealed that cases featuring males out-numbered those featuring females; this ratio differed across courses, and appeared to differ from the actual epidemiology of the conditions. Sexual orientation was specified infrequently. When sexual orientation and behavior were specified, these appeared in the context of a risk assessment for particular diseases (e.g., HIV infection). Most cases did not provide racial or ethnic descriptions. For many of the ethnic descriptors, links to genetic, cultural, or socioeconomic factors were apparent; no such link was apparent when the racial terms "white" or "Caucasian" were used. Analysis of the 983 cases shows that the pattern of demographics and associations of particular groups with diseases or risk factors in cases conveys messages, as does the lack of mention of sexual orientation and race or ethnicity. These messages are inconsistent with and may undermine the formal multicultural medical curriculum. The results suggest a need for formal deliberation of this aspect of the curriculum by curriculum planners. PMID- 11891161 TI - Medicine and the arts. Whoever was using this bed. PMID- 11891163 TI - Team communications in the operating room: talk patterns, sites of tension, and implications for novices. AB - PURPOSE: Although the communication that occurs within health care teams is important to both team function and the socialization of novices, the nature of team communication and its educational influence are not well documented. This study explored the nature of communications among operating room (OR) team members from surgery, nursing, and anesthesia to identify common communicative patterns, sites of tension, and their impact on novices. METHOD: Paired researchers observed 128 hours of OR interactions during 35 procedures from four surgical divisions at one teaching hospital. Brief, unstructured interviews were conducted following each observation. Field notes were independently read by each researcher and coded for emergent themes in the grounded theory tradition. Coding consensus was achieved via regular discussion. Findings were returned to insider "experts" for their assessment of authenticity and adequacy. RESULTS: Patterns of communication were complex and socially motivated. Dominant themes were time, safety and sterility, resources, roles, and situation. Communicative tension arose regularly in relation to these themes. Each procedure had one to four "higher-tension" events, which often had a ripple effect, spreading tension to other participants and contexts. Surgical trainees responded to tension by withdrawing from the communication or mimicking the senior staff surgeon. Both responses had negative implications for their own team relations. CONCLUSIONS: Team communications in the OR follow observable patterns and are influenced by recurrent themes that suggest sites of team tension. Tension in team communication affects novices, who respond with behaviors that may intensify rather than resolve interprofessional conflict. PMID- 11891164 TI - Determining the predictors of internal medicine residency accreditation: what they do (not what they say). AB - PURPOSE: The Accreditation Council for Graduate Medical Education and the Residency Review Committee for Internal Medicine (RRC-IM) evaluate internal medicine residency programs using a list of 301 program requirements. The authors investigated which requirements, program demographics, and site-visitor characteristics were the strongest predictors of accreditation. METHOD: The authors surveyed the program directors of all 405 accredited internal medicine residency programs in February 1998, obtaining data on the duration of the accreditation process, site visitors, and number and quality of citations. They also requested a copy of the notification letter containing citations and length of time until the next accreditation site visit (cycle length). RESULTS: A total of 217 responses (54%) was received. The mean cycle length was 3.0 years, and the accreditation process averaged 14.5 months. Smaller programs had a shorter average cycle length. Site visitors were reported to be prepared and professional overall. However, site visitors with the lowest evaluations by program directors were associated with shorter cycle lengths. Four program characteristics and program citations accounted for 60% of the variation in cycle length: total number of citations in the notification letter, percentage of graduates passing the American Board of Internal Medicine Certifying Examination, inadequate demonstration of resident scholarship, and inadequate ambulatory care experience. CONCLUSION: The authors devised an independent mechanism for determining the duration of the RRC-IM review process, influence of program demographics on the process, influence of site visitors on the accreditation action, and program requirements having the greatest effect on cycle length. PMID- 11891165 TI - Outcomes of combined internal medicine-pediatrics residency programs: a review of the literature. AB - PURPOSE: To evaluate the outcomes of combined internal medicine and pediatrics (IMP) residency programs using the published literature. METHOD: A literature search was conducted using Medline. Published articles were categorized as research or non-research (commentary, program requirements, program descriptions). Research articles were summarized and the results grouped under ten outcome variables. RESULTS: Of the 32 articles located, 18 were research articles, of which only four had been published since 1993. All the research conducted was cross-sectional and most involved surveys of program directors (seven studies) or graduates (four studies). At the time the studies were conducted, 20-33% of IMP residents did not complete their combined training; attrition rates have not been documented recently. Approximately 80% of IMP graduates achieved certification in both specialties, one third subspecialized to some degree, and 80% provided care to both adults and children. One fourth of the graduates felt that more training was needed in ambulatory settings and less was needed in intensive care. Very few studies of the outcomes of IMP physicians were found. CONCLUSION: The outcomes of IMP residency programs are important for health workforce policy, and this study documents a need for more extensive research on the outcomes of training programs for primary care physicians. PMID- 11891166 TI - The medical condition regard scale: measuring reactions to diagnoses. AB - PURPOSE: To develop a non-condition-specific scale to capture biases, emotions, and expectations generated by medical condition descriptors. METHOD: An 18-item pilot scale was developed from the literature on physicians' responses to patients they like and dislike, stigma definitions, and discussions with primary care faculty. Exploratory factor analysis was conducted after 440 medical students rated one of 12 diverse conditions. Confirmatory factor analysis was performed after 163 medical students rated two psychiatric conditions. Validity was evaluated by the scale's ability to meaningfully stratify the 12 conditions and identify changes in attitudes toward psychiatric conditions after a psychiatry clerkship. RESULTS: Exploratory factor analysis supported an 11-item unidimensional solution (all factor loadings >.40, coefficient alpha =.87). The final scale, the Medical Condition Regard Scale (MCRS), taps the degree to which medical students find patients with a given medical condition to be enjoyable, treatable, and worthy of medical resources. The unidimensional model also was supported by the confirmatory factor analyses for the two psychiatric conditions (both comparative fit indices =.98). The scale stratified the 12 conditions as expected: straightforward medical conditions rated highest, somatoform conditions rated lowest. Students showed greater regard for patients with major depression after the psychiatry clerkship, and students who rotated through an addiction treatment program showed a greater increase in regard for patients with alcoholism than did students not exposed to addiction treatment. CONCLUSION: MCRS scores are reliable, and the scale appears to be a valid instrument for assessing regard for any medical condition. PMID- 11891167 TI - An interactive, web-based tool for learning anatomic landmarks. AB - PURPOSE: To evaluate the effectiveness of a Web-based interactive teaching tool that uses self-assessment exercises with real-time feedback to aid students' learning in a gross anatomy class. METHOD: A total of 107 of 124 first-year medical students at one school were enrolled in the study. Students were divided into three groups: Group 1 (n = 63) received introductory material and activated their Web-based accounts; Group 2 (n = 44) received introductory material but did not activate their Web-based accounts; and Group 3 (n = 17) were not enrolled in the study and received no introductory material. Students in Group 1 had access to a graphic showing the locations of anatomic landmarks, a drill exercise, and a self-evaluation exercise. Students' ability to identify the anatomic landmarks on a 30-question midterm and a 30-question final exam were compared among the groups. RESULTS: The mean scores of students in Group 1 (midterm = 28.5, final = 28.1) were significantly higher than were the mean scores of students in Group 2 (midterm = 26.8, p <.001; final = 26.9, p <.017) and Group 3 (midterm = 24.8, p <.001; final = 26.4, p <.007). CONCLUSIONS: The Web-based tool was effective in improving students' scores on anatomic landmark exams. Future studies will determine whether the tool aids students in identifying structures located in three-dimensional space within regions such as the cranium and the abdominal cavity. PMID- 11891168 TI - Effect of publication bias on retrieval bias. AB - Publication bias is the main etiologic factor in retrieval bias. The authors measured the influence a positive study outcome had on housestaff's selecting the study for presentation. PMID- 11891169 TI - Feedback process between faculty and students. AB - The authors assessed-feedback agreement scores designed to measure the effectiveness of feedback between faculty and students in clinical medicine. PMID- 11891170 TI - A clinical-performance biopsy instrument. AB - A clinical-performance biopsy (CPB) instrument was introduced to enhance direct observation and timely feedback from supervising faculty during family practice resident continuity clinics. PMID- 11891172 TI - How many mutations in a cancer? PMID- 11891173 TI - Detection of BCL2 rearrangements in follicular lymphoma. PMID- 11891174 TI - Interstitial fibroblast-like cells express renin-angiotensin system components in a fibrosing murine kidney. AB - Recently, the renin-angiotensin system (RAS) was implicated in organ fibrosis. However, few studies have examined the localization of RAS components, such as angiotensin II receptors, renin (REN), angiotensinogen (AGTN), and angiotensin converting enzyme (ACE), in the fibrosing kidney. To localize these components in the fibrosing kidney, we used a murine model of renal fibrosis that shows an enhanced expression of angiotensin II type 1A receptor (AT(1A)R) and AGTN. Our results indicate that the overall expression of angiotensin II type 2 receptor (AT(2)R) and ACE was attenuated in this model, whereas REN expression was unchanged. In addition to tubular epithelial cells that were positive for AT(1A)R, AT(2)R, REN, and AGTN, interstitial fibroblast-like cells expressed AT(1A)R, REN, AGTN, and ACE in the fibrosing kidney. The interstitial fibroblast like cells that were positive for AT(1A)R mRNA were further characterized as positive for the expression of vimentin and transforming growth factor-beta1. These data provide strong evidence for a tubulointerstitial RAS within the fibrosing kidney, and a linkage between the RAS and renal fibrogenesis. PMID- 11891175 TI - Down-regulation of angiopoietin-1 expression in menorrhagia. AB - Angiogenesis is an essential component of endometrial repair and regeneration following menses. Perturbation of this process is associated with menorrhagia, a common gynecological disorder that results in excessive menstrual bleeding. Angiopoietin-1 (Ang-1) promotes vascular maturation via the Tie-2 receptor, while angiopoietin-2 (Ang-2) is its natural antagonist that destabilizes vessels and initiates neovascularization in the presence of vascular endothelial growth factor. To test the hypothesis that menorrhagia arises as a result of poor signal for vascular maturation, we have examined the expression of Ang-1, Ang-2, and Tie 2 in endometrium throughout the menstrual cycle from 30 normal women and 28 patients with menorrhagia. Ribonuclease protection assay and Western blot analysis showed Ang-2 expression was consistently higher than Ang-1 in normal endometrium throughout the cycle. However, with menorrhagia Ang-1 mRNA and protein were not detected or down-regulated, while Ang-2 was observed at similar levels in both normal and menorrhagic endometrium resulting in a greater than a 50% decrease in the ratio of Ang-1 to Ang-2 protein. In situ hybridization and immunohistochemical studies supported these findings and revealed cyclical changes in the expression of Ang-1 and Ang-2. These results suggest that the angiopoietin/Tie-2 system promotes vascular remodeling in endometrium and loss of normal Ang-1 expression may contribute to the excessive blood loss observed in menorrhagia. PMID- 11891176 TI - A defective, rearranged Epstein-Barr virus genome in EBER-negative and EBER positive Hodgkin's disease. AB - A ubiquitous herpesvirus that establishes life-long infection, the Epstein-Barr virus (EBV) has yielded little insight into how a single agent in general accord with its host can produce diverse pathologies ranging from oral hairy leukoplakia to nasopharyngeal carcinoma, from infectious mononucleosis to Hodgkin's disease (HD) and Burkitt's lymphoma. Its pathogenesis is further confounded by the less than total association of virus with histologically similar tumors. In other viral systems, defective (interfering) viral genomes are known to modulate outcome of infection, with either ameliorating or intensifying effects on disease processes initiated by prototype strains. To ascertain whether defective EBV genomes are present in HD, we examined paraffin-embedded tissue from 56 HD cases whose EBV status was first determined by cytohybridization for nonpolyadenylated EBV RNAs (EBERs). Using both standard polymerase chain reaction (PCR) and PCR in situ hybridization, we successfully amplified sequences that span abnormally juxtaposed BamHI W and Z fragments characteristic of defective heterogeneous (het) EBV DNA from 10 of 32 (31%) EBER-positive tumors. Of 24 EBER-negative HD, 8 yielded PCR products indicating presence of het EBV DNA. Two of these contained defective EBV in the apparent absence of the prototype virus. Of the 42 tumors analyzed for defective EBV by both PCR techniques, there was concordance of results in 38 (90%). Detection of defective EBV genomes with the potential to disrupt viral gene regulation suggests one mechanism for pathogenic diversity that may also account for loss of prototypic EBV from individual tumor cells. PMID- 11891177 TI - Epstein-barr virus-positive gastric carcinoma demonstrates frequent aberrant methylation of multiple genes and constitutes CpG island methylator phenotype positive gastric carcinoma. AB - CpG island methylation is an important mechanism for inactivating the genes involved in tumorigenesis. Gastric carcinoma (GC) is one of the tumors that exhibits a high frequency of aberrant CpG island methylation. There have been many reports suggesting a close link between Epstein-Barr virus (EBV) and the development of GC. However, little is known about the oncogenic mechanism of EBV in gastric carcinogenesis. Twenty-one cases of EBV-positive GC and 56 cases of EBV-negative GC were examined for aberrant DNA methylation of the CpG islands of 19 genes or loci and the differences in the methylation frequency between EBV positive and -negative GCs were investigated to determine a role of aberrant methylation in EBV-related gastric carcinogenesis. The average number of methylated genes or loci was higher in EBV-positive GCs than in EBV-negative GCs (13.4 versus 7.8, respectively, P < 0.001). EBV-positive GCs showed methylation in at least 10 CpG islands (52.6% of the tested genes), whereas 62.5% of EBV negative GCs showed methylation in <10 CpG islands. THBS1, APC, p16, 14-3-3 sigma, MINT1, and MINT25 were methylated at a frequency >90% in EBV-positive GCs. The methylation frequency difference in the respective CpG islands between EBV positive and -negative GCs was statistically significant (P < 0.05). Among these genes or loci, the methylation frequency of p16 in the EBV-positive GCs was more than three times higher than in the EBV-negative GCs. The PTEN, RASSF1A, GSTP1, MGMT, and MINT2 were methylated in EBV-positive GCs at a frequency of more than three times that of the EBV-negative GCs. These results demonstrate a relationship between EBV and aberrant methylation in GC and suggest that aberrant methylation may be an important mechanism of EBV-related gastric carcinogenesis. PMID- 11891178 TI - Considerations when analyzing the methylation status of PTEN tumor suppressor gene. AB - Epigenetic mechanisms of gene silencing, including promoter hypermethylation of tumor suppressor genes, have been shown to contribute to tumorigenesis. PTEN is an important tumor suppressor implicated in the pathogenesis of a number of familial and sporadic cancers. Germline mutations of PTEN predispose to dominantly inherited hamartomatous disorders Cowden syndrome and Bannayan-Zonana syndrome. Somatic PTEN mutations commonly occur in endometrial, breast, prostate, and thyroid tumors. Several investigators have speculated on PTEN promoter hypermethylation as a possible mechanism of PTEN inactivation but data supporting such observations is not forthcoming. The genomic sequence of PTEN is 98% identical to a highly conserved processed PTEN pseudogene (psiPTEN) and this sequence identity extends 841 base pairs into the promoter region. This high degree of homology has made analysis of the methylation status of the PTEN promoter quite challenging. We have investigated the methylation profiles of the promoter region of PTEN in endometrial, breast, and colon cancer cell lines, as well as in a panel of primary endometrial tumors using a combination of methylation-specific polymerase chain reaction, methylation-sensitive restriction analysis, and bisulfite sequencing. Our results show that the pseudogene, and not PTEN, is predominantly methylated in cell lines and tumors. Without careful consideration of the critical nucleotide differences between the two sequences, results obtained from PTEN analysis may not necessarily represent the methylation status of PTEN. PMID- 11891179 TI - mRNA expression profiling of laser microbeam microdissected cells from slender embryonic structures. AB - Microarray hybridization has rapidly evolved as an important tool for genomic studies and studies of gene regulation at the transcriptome level. Expression profiles from homogenous samples such as yeast and mammalian cell cultures are currently extending our understanding of biology, whereas analyses of multicellular organisms are more difficult because of tissue complexity. The combination of laser microdissection, RNA amplification, and microarray hybridization has the potential to provide expression profiles from selected populations of cells in vivo. In this article, we present and evaluate an experimental procedure for global gene expression analysis of slender embryonic structures using laser microbeam microdissection and laser pressure catapulting. As a proof of principle, expression profiles from 1000 cells in the mouse embryonic (E9.5) dorsal aorta were generated and compared with profiles for captured mesenchymal cells located one cell diameter further away from the aortic lumen. A number of genes were overexpressed in the aorta, including 11 previously known markers for blood vessels. Among the blood vessel markers were endoglin, tie-2, PDGFB, and integrin-beta1, that are important regulators of blood vessel formation. This demonstrates that microarray analysis of laser microbeam micro dissected cells is sufficiently sensitive for identifying genes with regulative functions. PMID- 11891180 TI - Laser capture microdissection and two-dimensional polyacrylamide gel electrophoresis: evaluation of tissue preparation and sample limitations. AB - Laser capture microdissection (LCM) is now well established as a tool for facilitating the enrichment of cells of interest from tissue sections, overcoming the problem of tissue heterogeneity. LCM has been used extensively in combination with analysis at the DNA and RNA levels, but only a small number of studies have employed LCM with subsequent protein analysis, albeit with promising results. This study focuses on the potential of LCM in combination with two-dimensional polyacrylamide gel electrophoresis. The effects of tissue section preparation and sample type were evaluated to fully determine the suitability of using LCM in global protein profiling. The effects of several histochemical stains (hematoxylin and eosin, methyl green and toluidine blue) and immunolabeling on subsequent two-dimensional polyacrylamide gel electrophoresis were investigated. Quantitative analysis was performed to establish the extent of changes in the relative intensity of protein species and their reproducibility. All staining protocols tested were found to be compatible with protein analysis although there was variation in protein recovery and the quality of the protein profiles obtained. LCM of renal and cervix samples indicated that protein yield after dissection was acceptable, although the extent of enrichment and dissection time was tissue-dependent, which may preclude the use of this approach with some tissue types. These results indicate that LCM has potential as a tool in proteomic research. PMID- 11891181 TI - High frequency of t(14;18)-translocation breakpoints outside of major breakpoint and minor cluster regions in follicular lymphomas: improved polymerase chain reaction protocols for their detection. AB - The detection of t(14;18) translocations is widely used for the diagnosis and monitoring of follicular lymphomas displaying a high prevalence for this aberration. Cytogenetics, Southern blotting, and polymerase chain reaction (PCR) are commonly used techniques. It is generally believed that the vast majority of the breakpoints occurs on chromosome 18 in the major breakpoint region (mbr) and the minor cluster region (mcr). Yet, by improving long-distance PCR protocols we identified half of the breakpoints outside of these clusters. Our study included biopsies from 59 patients with follicular lymphoma. Seventy-one percent carried translocations detectable with our long-distance PCR protocol. The novel primer sets were derived from the hitherto uncharacterized 25-kb-long stretch between mbr and mcr that we have sequenced for this purpose. Sequence analysis of the novel breakpoints reveals a wide distribution between mbr and mcr displaying some clustering 16 kb downstream from the BCL2 gene. By including a primer for this intermediate cluster region in standard PCRs we could also improve the detection of t(14;18) translocations in formalin-fixed and paraffin-embedded biopsies. Our new PCRs are highly sensitive, easy to perform, and thus well suited for routine analysis of t(14;18) translocations for the primary diagnosis of follicular lymphoma and surveillance of minimal residual disease. PMID- 11891182 TI - Up-regulation of MHC class I expression accompanies but is not required for spontaneous myopathy in dysferlin-deficient SJL/J mice. AB - We found that up-regulation of major histocompatibility complex (MHC) class I expression accompanies, but is not required for, appearance of spontaneous myopathy in SJL/J mice. In some neuromuscular diseases, MHC class I expression is markedly up-regulated in muscles, though the consequences of this up-regulation for pathology are not clear. To study MHC class I in myopathy, we compared muscles of SJL/J mice to muscles of SJL/J mice that were also MHC class I deficient due to targeted mutation in the beta-2-microglobulin gene (SJL/J B2m ( /-) mice). SJL/J mice show spontaneous myopathy and have a mutation in the dysferlin gene, a gene which is also mutated in human limb-girdle muscular dystrophy type 2B (LGMD2B). Muscles of eight-month-old SJL/J mice had higher levels of MHC class I expression than muscles of either C57BL/6J (wild-type) or SJL/J B2m (-/-) mice. In contrast, the percentage of abnormal muscle fibers was similar in SJL/J and SJL/J B2m (-/-) muscles. Invading Mac-1(+) cells were most abundant in SJL/J B2m (-/-) muscles, moderately abundant in SJL/J muscles, and rare in C57BL/6J muscles. Thus, MHC class I was markedly up-regulated in SJL/J muscles, but this high level of MHC class I was not necessary for the appearance of myopathy. PMID- 11891183 TI - CAP37, a novel inflammatory mediator: its expression in endothelial cells and localization to atherosclerotic lesions. AB - Cationic antimicrobial protein of 37 kd (CAP37), originally isolated from human neutrophils, is an important multifunctional inflammatory mediator. Here we describe its localization within the vascular endothelium associated with atherosclerotic plaques. Evidence from in vitro immunocytochemical, Northern blot, and reverse transcriptase-polymerase chain reaction analysis indicates that CAP37 is induced in endothelial cells in response to inflammatory mediators. Endothelial-derived CAP37 shows sequence identity with an extensive region of neutrophil-derived CAP37. This is the first demonstration of endogenous endothelial CAP37, confirmed by sequence analysis. We suggest that, because of its induction and location in the endothelium and its known monocyte- and endothelial-activating capabilities, CAP37 has potential to modulate monocyte/endothelial dynamics at the vessel wall in inflammation. PMID- 11891184 TI - The expression of ccn3(nov) gene in musculoskeletal tumors. AB - The CCN3(NOV) protein belongs to the CCN [cysteine-rich CYR61, connective tissue growth factor (CTGF), nephroblastoma overexpressed gene (Nov)] family of growth regulators, sharing a strikingly conserved multimodular organization but exhibiting distinctive functional features. Although previous studies have revealed an expression of CCN3 protein in several normal tissues, including kidney, nervous system, lung, muscle, and cartilage, less is known about its expression in tumors. In this study, we analyzed the expression of CCN3 in musculoskeletal tumors, using a panel of human cell lines and tissue samples. An association between CCN3 expression and tumor differentiation was observed in rhabdomyosarcoma and cartilage tumors, whereas, in Ewing's sarcoma, the expression of this protein seemed to be associated with a higher risk to develop metastases. CCN3 expression was found in 15 of 45 Ewing's sarcoma tissue samples. In particular, we did not observe any expression of CCN3 in the 15 primary tumors that did not develop metastases. In contrast, 15 of the 30 primary tumors that developed lung and/or bone metachronous metastases showed a high expression of the protein (P < 0.001, Fisher's test). Our studies indicate that CCN3 is generally expressed in the cells of the musculoskeletal system. This protein may play a role both in normal and pathological conditions. However, the regulation of CCN3 expression varies in the different neoplasms and depends on the type of cells. Thus, as reported for other CCN genes, the biological properties and regulation of expression of CCN3 are dependent on the cellular context and the nature of the cells in which it is produced. Further studies will help to clarify the biological role of this protein in musculoskeletal neoplasms. PMID- 11891185 TI - The alpha8 integrin chain affords mechanical stability to the glomerular capillary tuft in hypertensive glomerular disease. AB - In the kidney, the alpha8 integrin chain is expressed in glomerular mesangial cells. The alpha8 integrin plays a role in early nephrogenesis but its functional role in the adult kidney is unknown. We tested the hypothesis that alpha8 integrin-mediated cell-matrix interactions are important to maintain the integrity of the glomerulus in arterial hypertension. Desoxycorticosterone (DOCA) salt hypertension was induced in mice homozygous for a deletion of the alpha8 integrin chain and wild-type mice. Blood pressure, albumin excretion, total renal mass, and glomerular filtration in DOCA-treated alpha8-deficient mice were comparable to DOCA-treated wild types. DOCA-treated wild types showed increased glomerular immunostaining for alpha8 integrin compared to salt-loaded and untreated controls, whereas the glomeruli of alpha8-deficient mice always stained negative. Morphometric studies revealed similar degrees of glomerulosclerosis in DOCA-treated alpha8-deficient and DOCA-treated wild-type mice. However, DOCA treated alpha8-deficient mice had a higher score of capillary widening (mesangiolysis) than DOCA-treated wild-type mice, which was confirmed in two additional wild-type strains. Moreover, in DOCA-treated alpha8-deficient mice, glomerular fibrin deposits were more frequent than in DOCA-treated wild types. The results show that lack of alpha8 is associated with increased susceptibility to glomerular capillary destruction in DOCA salt hypertension, whereas it does not seem to play a major role in the development of fibrosis or glomerulosclerosis. Our findings indicate that mesangial alpha8 integrin contributes to maintain the integrity of the glomerular capillary tuft during mechanical stress, eg, in hypertension. PMID- 11891186 TI - Thymosin-beta4 regulates motility and metastasis of malignant mouse fibrosarcoma cells. AB - We identified a thymosin-beta4 gene overexpression in malignant mouse fibrosarcoma cells (QRsP-30) that were derived from clonal weakly tumorigenic and nonmetastatic QR-32 cells by using a differential display method. Thymosin-beta4 is known as a 4.9-kd polypeptide that interacts with G-actin and functions as a major actin-sequestering protein in cells. All of the six malignant fibrosarcoma cell lines that have been independently converted from QR-32 cells expressed high levels of thymosin-beta4 mRNA and its expression in tumor cells was correlated with tumorigenicity and metastatic potential. Up-regulation of thymosin-beta4 in QR-32 cells (32-S) transfected with sense thymosin-beta4 cDNA converted the cells to develop tumors and formed numerous lung metastases in syngeneic C57BL/6 mice. In contrast, antisense thymosin-beta4 cDNA-transfected QRsP-30 (30-AS) cells reduced thymosin-beta4 expression, and significantly lost tumor formation and metastases to distant organs. Vector-alone transfected cells (32-V or 30-V cells) behaved like their parental cells. We observed that tumor cell motility, cell shape, and F-actin organization is regulated in proportion to the level of thymosin-beta4 expression. These findings indicate that thymosin-beta4 molecule regulates fibrosarcoma cell tumorigenicity and metastasis through actin-based cytoskeletal organization. PMID- 11891187 TI - In situ analysis of the variable heavy chain gene of an IgM/IgG-expressing follicular lymphoma: evidence for interfollicular trafficking of tumor cells. AB - It is generally assumed that follicular lymphomas (FL) not only morphologically resemble normal germinal centers but have retained some functional characteristics of their non-neoplastic counterparts as well. Recent IgV gene analyses on a panel of FLs however, strongly suggested that FLs do not retain the capacity of somatic hypermutation and are not being selected on basis of the quality of their mIgV regions. To extend these findings, we investigated the follicular organization and class switching in a FL that consisted of both IgM- and IgG-expressing tumor cells with a high somatic mutation load and significant intraclonal V(H) gene diversity. V(H)-C(mu) and V(H)-Cgamma gene transcripts were amplified and sequenced from samples of approximately 50 tumor cells, isolated from frozen tissue sections by laser microdissection. We identified many different subclones and obtained limited evidence of subclone dominance in individual follicles. Remarkably, several subclones were found scattered over different follicles. All samples contained IgM- and IgG-expressing tumor cells with, in general, non-identical mutation patterns, which is not in support of ongoing class switching. Accordingly, no switch circle recombination products were found. The findings indicate that the neoplastic follicles lack the organization and functions typical of reactive germinal centers. PMID- 11891188 TI - Expression of cyclooxygenase 2 is an independent prognostic factor in human ovarian carcinoma. AB - Cyclooxygenase-2 (COX-2) is the rate-limiting enzyme in prostanoid biosynthesis and is involved in tumor progression. We investigated expression of COX-1 and COX 2 in cell lines and tumors from ovarian carcinomas. Expression of COX-2 mRNA and protein was detectable in three of five ovarian carcinoma cell lines and was inducible by interleukin-1beta or phorbolester in a subset of cell lines. Prostaglandin E(2) (PGE(2)) production could be inhibited by the selective COX-2 inhibitor NS-398. In malignant ascites of ovarian carcinomas significantly increased levels of PGE(2) were found compared to other carcinomas or nonmalignant ascites (P = 0.03). We investigated expression of COX-2 by immunohistochemistry in 117 ovarian surface epithelial tumors. Expression of COX 2 was detected in 42% of 86 ovarian carcinomas and in 37% of 19 low malignant potential tumors, but not in 12 cystadenomas or 2 normal ovaries. Expression of COX-1 was detected by immunohistochemistry in 75% of 75 invasive ovarian carcinomas and in 75% of 16 low malignant potential tumors, whereas 2 samples from normal ovaries and 8 cystadenomas were positive for COX-1. In univariate survival analysis of invasive carcinomas, expression of COX-2 was associated with a significantly reduced median survival time (log rank test, P = 0.04). For patients younger than 60 years of age, this association was even more significant (P < 0.004). In contrast, expression of COX-1 was no prognostic parameter (P = 0.89). There was no significant correlation between COX-2 or COX-1 expression and other clinicopathological markers. In multivariate analysis expression of COX-2 was an independent prognostic factor for poor survival (relative risk, 2.74; 95% CI, 1.38 to 5.47). Our data indicate that COX-2 expression is an independent prognostic factor in ovarian carcinoma. Based on the results of this study, it would be interesting to investigate whether ovarian carcinoma patients with tumors positive for COX-2 would benefit from treatment with selective COX-2 inhibitors. PMID- 11891189 TI - Nitric oxide mediates apoptosis through formation of peroxynitrite and Fas/Fas ligand interactions in experimental autoimmune uveitis. AB - Conflicting reports have led to the description of nitric oxide as a "double edged sword" in animal models of autoimmunity. In this study we show that tissue damage within the eye during experimental autoimmune uveoretinitis correlates with peroxynitrite formation in infiltrating monocytes/macrophages within the outer retina together with extensive photoreceptor apoptosis and apoptosis of Fas(+) T cells within the retina. Inducible nitric oxide synthase (NOS2) expression was primarily restricted to infiltrating monocytes/macrophages in the outer retina and photoreceptor rod outer segments (target tissue), but despite showing evidence of lipid peroxidation, myeloid cells remained resistant to apoptosis. The protective effect of the NOS inhibitor N(G)-nitro-L-arginine methyl ester could be attributed to dramatically reduced photoreceptor apoptosis, absence of nitrotyrosine formation, and reduced NOS2 protein expression. However, inhibition of NOS by N(G)-nitro-L-arginine methyl ester was accompanied by a sparing of CD3(+) and CD2(+) T cells despite continued expression of Fas and Fas ligand, thus compromising functional inactivation of T cells in the target tissue. These data suggests that in addition to contributing to tissue damage in the retina through generation of reactive oxygen species, nitric oxide also seems to be required for activation-induced cell death and elimination of T cells in the retina. PMID- 11891190 TI - Differing roles for urokinase and tissue-type plasminogen activator in collagen induced arthritis. AB - The plasminogen activators, urokinase PA (u-PA) and tissue-type PA (t-PA), are believed to play important roles in inflammatory cell infiltration, fibrin deposition, and joint destruction associated with rheumatoid arthritis; however, their precise roles in such processes, particularly u-PA, have yet to be defined. Using gene-deficient mice we examined the relative contribution of the PAs to the chronic systemic collagen-induced arthritis model. Based on clinical and histological assessments, u-PA-/- mice developed significantly milder disease and t-PA-/- mice more severe disease compared with the relevant wild-type mice. Fibrin deposition within joints paralleled disease severity and was particularly pronounced in t-PA-/- mice. Likewise, cytokine levels in the synovium reflected the severity of disease, with interleukin-1beta levels in particular being lower in u-PA-/- mice and increased in t-PA-/- mice. The antibody response to type II collagen was normal in both knockouts; however, T cells from u-PA-/- mice had a reduced proliferative response and produced less interferon-gamma on antigen stimulation in vitro. These results indicate that the major effect of u-PA in the collagen-induced arthritis model is deleterious, whereas that of t-PA is protective. Our data highlight the complexities of PA function, and suggest that approaches either to target u-PA or to enhance local t-PA activity in joints may be of therapeutic benefit in rheumatoid arthritis. PMID- 11891191 TI - Macrophages relate presynaptic and postsynaptic damage in simian immunodeficiency virus encephalitis. AB - Neurodegeneration observed in lentiviral-associated encephalitis has been linked to viral-infected and -activated central nervous system macrophages. We hypothesized that lentivirus, macrophages, or both lentivirus and macrophages within distinct microenvironments mediate synaptic damage. Using the simian immunodeficiency virus (SIV)-infected macaque model, we assessed the relationship between virus, macrophages, and neurological damage in multiple brain regions using laser confocal microscopy. In SIV-infected macaques with SIV encephalitis (SIVE), brain tissue concentrations of SIV RNA were 5 orders of magnitude greater than that observed in nonencephalitic animals. In SIVE, staining for postsynaptic protein microtubule-associated protein-2 was significantly decreased in the caudate, hippocampus, and frontal cortical gray matter compared to nonencephalitic controls, whereas staining for presynaptic protein synaptophysin was decreased in SIV-infected macaques with and without encephalitis. These data suggest that presynaptic damage occurs independent of pathological changes associated with SIVE, whereas postsynaptic damage is more tightly linked to regional presence of both activated and infected macrophages. PMID- 11891192 TI - Extensive induction of important mediators of fibrosis and dystrophic calcification in desmin-deficient cardiomyopathy. AB - Mice lacking the intermediate filament protein desmin demonstrate abnormal mitochondria behavior, disruption of muscle architecture, and myocardial degeneration with extensive calcium deposits and fibrosis. These abnormalities are associated with cardiomyocyte hypertrophy, cardiac chamber dilation and eventually with heart failure. In an effort to elucidate the molecular mechanisms leading to the observed pathogenesis, we have analyzed gene expression changes in cardiac tissue using differential display polymerase chain reaction and cDNA atlas array methods. The most substantial changes were found in genes coding the small extracellular matrix proteins osteopontin and decorin that are dramatically induced in the desmin-null myocardium. We further analyzed their expression pattern both at the RNA and protein levels and we compared their spatial expression with the onset of calcification. Extensive osteopontin localization is observed by immunohistochemistry in the desmin-null myocardium in areas with massive myocyte death, as well as in hypercellular regions with variable degrees of calcification and fibrosis. Osteopontin is consistently co-localized with calcified deposits, which progressively are transformed to psammoma bodies surrounded by decorin, especially in the right ventricle. These data together with the observed up-regulation of transforming growth factor-beta1 and angiotensin-converting enzyme, could explain the extensive fibrosis and dystrophic calcification observed in the heart of desmin-null mice, potentially crucial events leading to heart failure. PMID- 11891193 TI - Genetic and immunohistochemical analysis of pancreatic acinar cell carcinoma: frequent allelic loss on chromosome 11p and alterations in the APC/beta-catenin pathway. AB - Acinar cell carcinomas (ACCs) are rare malignant tumors of the exocrine pancreas. The specific molecular alterations that characterize ACCs have not yet been elucidated. ACCs are morphologically and genetically distinct from the more common pancreatic ductal adenocarcinomas. Instead, the morphological, immunohistochemical, and clinical features of ACCs overlap with those of another rare pancreatic neoplasm, pancreatoblastoma. We have recently demonstrated a high frequency of allelic loss on chromosome arm 11p and mutations in the APC/beta catenin pathway in pancreatoblastomas, suggesting that similar alterations might also play a role in the pathogenesis of some ACCs. We analyzed a series of 21 ACCs for somatic alterations in the APC/beta-catenin pathway and for allelic loss on chromosome 11p. In addition, we evaluated the ACCs for alterations in p53 and Dpc4 expression using immunohistochemistry, and for microsatellite instability (MSI) using polymerase chain amplification of a panel of microsatellite markers. Allelic loss on chromosome 11p was the most common genetic alteration in ACCs, present in 50% (6 of 12 informative cases). Molecular alterations in the APC/beta catenin pathway were detected in 23.5% (4 of 17) of the carcinomas, including one ACC with an activating mutation of the beta-catenin oncogene and three ACCs with truncating APC mutations. One ACC (1 of 13, 7.6%) showed allelic shifts in four of the five markers tested (MSI-high), two (15.4%) showed an allelic shift in only one of the five markers tested (MSI-low), and no shifts were detected in the remaining 10 cases. The MSI-high ACC showed medullary histological features. In contrast, no loss of Dpc4 protein expression or p53 accumulation was detected. These results indicate that ACCs are genetically distinct from pancreatic ductal adenocarcinomas, but some cases contain genetic alterations common to histologically similar pancreatoblastomas. PMID- 11891194 TI - Selective cyclooxygenase-2 blocker delays healing of esophageal ulcers in rats and inhibits ulceration-triggered c-Met/hepatocyte growth factor receptor induction and extracellular signal-regulated kinase 2 activation. AB - Nonsteroidal anti-inflammatory drugs, both nonselective and cyclooxygenase-2 (COX 2) selective, delay gastric ulcer healing. Whether they affect esophageal ulcer healing remains unexplored. We studied the effects of the COX-2 selective inhibitor, celecoxib, on esophageal ulcer healing as well as on the cellular and molecular events involved in the healing process. Esophageal ulcers were induced in rats by focal application of acetic acid. Rats with esophageal ulcers were treated intragastrically with either celecoxib (10 mg/kg, once daily) or vehicle for 2 or 4 days. Esophageal ulceration triggered increases in: esophageal epithelial cell proliferation; expression of COX-2 (but not COX-1); hepatocyte growth factor (HGF) and its receptor, c-Met; and activation of extracellular signal-regulated kinase 2 (ERK2). Treatment with celecoxib significantly delayed esophageal ulcer healing and suppressed ulceration-triggered increases in esophageal epithelial cell proliferation, c-Met mRNA and protein expression, and ERK2 activity. In an ex vivo organ-culture system, exogenous HGF significantly increased ERK2 phosphorylation levels in esophageal mucosa. A structural analog of celecoxib, SC-236, completely prevented this effect. These findings indicate that celecoxib delays esophageal ulcer healing by reducing ulceration-induced esophageal epithelial cell proliferation. These actions are associated with, and likely mediated by, down-regulation of the HGF/c-Met-ERK2 signaling pathway. PMID- 11891195 TI - Expression of PKD1 and PKD2 transcripts and proteins in human embryo and during normal kidney development. AB - Autosomal-dominant polycystic kidney disease, one of the most frequent human genetic disorders, is genetically heterogeneous. Most cases result from mutations of PKD1 or PKD2 encoding polycystin-1 or polycystin-2, respectively. Polycystin-1 is a large transmembrane protein containing several domains involved in cell-cell and/or cell-matrix interactions. Polycystin-2 is transmembrane glycoprotein sharing homology with some families of cation channels. Despite a large number of reports, the tissue distribution of these two proteins, especially of polycystin 1, is still debated. We investigated the expression pattern of PKD1 and PKD2 transcripts and proteins during human embryogenesis and kidney development, using Northern blot analysis, in situ hybridization, and immunohistochemical methods. For each gene, the expression pattern of transcripts and protein was concordant. In human 5- to 6-week-old embryos, both genes are widely expressed, mainly in neural tissue, cardiomyocytes, endodermal derivatives, and mesonephros. At this age, PKD2 but not PKD1 expression is observed in the ureteric bud and the uninduced metanephros. Thereafter, PKD2 is diffusely expressed at all stages of nephron development, whereas high PKD1 expression first appears in differentiated proximal tubules. Proximal tubule expression of both genes decreases from weeks 20 to 24 onwards. PKD1 transcripts, later restricted to distal tubules in fetal nephrogenesis, are no longer detected in adult kidneys, which nevertheless maintain a faint expression of polycystin-1, whereas persistent expression of PKD2 transcripts and protein is observed throughout nephrogenesis. Overall, contrary to previous observations, we found profound differences in the spatiotemporal expression of PKD1 and PKD2 during nephrogenesis, PKD2 being expressed earlier and more diffusely than PKD1. These data suggest that polycystins could interact with different partners, at least during kidney development. PMID- 11891197 TI - Lectin histochemistry of resected adenocarcinoma of the lung: helix pomatia agglutinin binding is an independent prognostic factor. AB - The worldwide incidence of adenocarcinoma of the lung is rising. Unfortunately, no significant prognostic marker beyond the classical TNM staging exists to stratify these patients for appropriate therapy. Because lectins, carbohydrate binding proteins, have been shown to be useful prognostic markers in several other adenocarcinomas, a panel of lectins [Helix pomatia agglutinin (HPA), Phaseolus vulgaris leukoagglutinin, Ulex europaeus agglutinin, Maackia amurenis agglutinin, Sambucus nigra agglutinin] with different carbohydrate-binding specificities were tested for their prognostic relevance. Paraffin wax sections of 93 patients with adenocarcinomas of the lung who had undergone surgery between 1990 and 1995 were investigated by lectin histochemistry. Lectin-binding data and other known prognostic factors were correlated with survival. In univariate analysis, binding of HPA, Phaseolus vulgaris leukoagglutinin, and Ulex europaeus agglutinin to adenocarcinoma cells were prognostic indicators for overall and relapse-free survival, whereas Maackia amurenis agglutinin and Sambucus nigra agglutinin binding had no prognostic value. However, in a multivariate analysis next to stage and gender only HPA was a significant independent prognostic factor on survival. In conclusion, HPA binding was the primary marker-based predictor of prognosis in our patient population and allows to stratify patients with adenocarcinomas of the lung into a low- and a high-risk group. PMID- 11891196 TI - Abnormalities in pericytes on blood vessels and endothelial sprouts in tumors. AB - Endothelial cells of tumor vessels have well-documented alterations, but it is less clear whether pericytes on these vessels are abnormal or even absent. Here we report that alpha-smooth muscle actin (alpha-SMA) and desmin-immunoreactive pericytes were present on >97% of blood vessels viewed by confocal microscopy in 100-microm-thick sections of three different spontaneous or implanted tumors in mice. However, the cells had multiple abnormalities. Unlike pericytes on capillaries in normal pancreatic islets, which had desmin but not alpha-SMA immunoreactivity, pericytes on capillary-size vessels in insulinomas in RIP-Tag2 transgenic mice expressed both desmin and alpha-SMA. Furthermore, pericytes in RIP-Tag2 tumors, as well as those in MCa-IV breast carcinomas and Lewis lung carcinomas, had an abnormally loose association with endothelial cells and extended cytoplasmic processes deep into the tumor tissue. alpha-SMA-positive pericytes also covered 73% of endothelial sprouts in RIP-Tag2 tumors and 92% of sprouts in the other tumors. Indeed, pericyte sleeves were significantly longer than the CD31-immunoreactive endothelial cell sprouts themselves in all three types of tumors. All three tumors also contained alpha-SMA-positive myofibroblasts that resembled pericytes but were not associated with blood vessels. We conclude that pericytes are present on most tumor vessels but have multiple abnormalities, including altered expression of marker proteins. In contrast to some previous studies, the almost ubiquitous presence of pericytes on tumor vessels found in the present study may be attributed to our use of both desmin and alpha-SMA as markers and 100-microm-thick tissue sections. The association of pericytes with endothelial sprouts raises the possibility of an involvement in sprout growth or retraction in tumors. PMID- 11891198 TI - Importance of vascular phenotype by basic fibroblast growth factor, and influence of the angiogenic factors basic fibroblast growth factor/fibroblast growth factor receptor-1 and ephrin-A1/EphA2 on melanoma progression. AB - The expression of several angiogenic factors and receptors was examined in a series of vertical growth phase cutaneous melanomas using high-throughput tissue microarray technology and immunohistochemistry. The results were correlated with microvessel density, clinicopathological features, and patient survival. Expression of basic fibroblast growth factor (bFGF) was significantly associated with increased microvessel density. Also, we found an independent prognostic importance of vascular phenotype by endothelial cell expression of bFGF; cases with positive vessels had the best prognosis and these tumors revealed a low frequency of vascular invasion (14%) when compared with bFGF-negative vessels (47%). This bFGF-negative phenotype was significantly increased in metastatic lesions. Strong tumor cell expression of FLT-4, ephrin-A1, and EphA2 was associated with increased melanoma thickness, and ephrin-A1 staining was related to decreased survival (P = 0.039). Expression of EphA2 in tumor cells was associated with increased tumor cell proliferation (Ki-67 positivity), indicating possible autocrine growth stimulation. Thus, our findings indicate the presence of phenotypic diversity among tumor-associated vessels, and subgroups defined by bFGF expression may be of clinical importance. bFGF was associated with microvessel density, whereas the ephrin-A1/EphA2 pathway might also be important for tumor cell proliferation and patient survival. PMID- 11891199 TI - Fibrinogen stabilizes placental-maternal attachment during embryonic development in the mouse. AB - In humans, maternal fibrinogen (Fg) is required to support pregnancies by maintaining hemostatic balance and stabilizing uteroplacental attachment at the fibrinoid layer found at the fetal-maternal junction. To examine relationships between low Fg levels and early fetal loss, a genetic model of afibrinogenemia was developed. Pregnant mice homozygous for a deletion of the Fg-gamma chain, which results in a total Fg deficiency state (FG(-/-)), aborted the fetuses at the equivalent gestational stage seen in humans. Results obtained from timed matings of FG(-/-) mice showed that vaginal bleeding was initiated as early as embryonic day (E)6 to 7, a critical stage for maternal-fetal vascular development. The condition of afibrinogenemia retarded embryo-placental development, and consistently led to abortion and maternal death at E9.75. Lack of Fg did not alter the extent or distribution pattern of other putative factors of embryo-placental attachment, including laminin, fibronectin, and Factor XIII, indicating that the presence of fibrin(ogen) is required to confer sufficient stability at the placental-decidual interface. The results of these studies demonstrate that maternal Fg plays a critical role in maintenance of pregnancy in mice, both by supporting proper development of fetal-maternal vascular communication and stabilization of embryo implantation. PMID- 11891200 TI - Monoclonal gamma-T-cell receptor rearrangement in vulvar lichen sclerosus and squamous cell carcinomas. AB - Risk factors for vulvar squamous cell carcinoma (SCC) are human papilloma virus (HPV) infections and lichen sclerosus (LS). The significance of monoclonal gamma T-cell receptor (gamma-TCR) rearrangement in the lymphoid infiltrate of LS and the consequence for vulvar carcinogenesis is unknown. One hundred sixty-one biopsies of vulvar LS and SCC, with and without LS, were examined for monoclonal gamma-TCR rearrangement and HPV16 expression, and for the expression of B- and T cell markers and fascin. Monoclonal gamma-TCR rearrangement was identified in 8 of 17 patients with LS and 11 of 21 patients with SCC arising in LS with only occasional HPV16 DNA detection. None of the 19 SCC without LS showed monoclonal gamma-TCR rearrangement, but 14 of 19 patients had strong HPV16 detection. The lichenoid infiltrate of LS with germline configuration consisted predominantly of T cells (CD8 > CD4), along with numerous B cells. However, in biopsies with monoclonally rearranged gamma-TCR, CD4-positive T cells dominated along with B cells and fascin-positive cells in the lichenoid infiltrate and in deeply located lymphocyte aggregates (LAs). These LAs additionally contained fascin-positive dendritic cells with only individual CD8, CD57, and granzyme-positive cells. LAs in biopsies with germline configuration demonstrated numerous T cells (CD8 >CD4), but only single peripheral B cells, CD57, and fascin-positive lymphocytes. Our data suggest that monoclonal gamma-TCR rearrangement is characteristic for and limited to LS and SCC arising in LS, raising the question for a LS-associated antigen. We interpret B cells, CD4-positive T cells, and fascin-expressing dendritic cells within LS as a cellular immune response to antigen or proliferating T-cell clones. The resulting local immune dysregulation in LS may provide a permissive environment for the development of a SCC. PMID- 11891201 TI - The metallothionein-null phenotype is associated with heightened sensitivity to lead toxicity and an inability to form inclusion bodies. AB - Susceptibility to lead toxicity in MT-null mice and cells, lacking the major forms of the metallothionein (MT) gene, was compared to wild-type (WT) mice or cells. Male MT-null and WT mice received lead in the drinking water (0 to 4000 ppm) for 10 to 20 weeks. Lead did not alter body weight in any group. Unlike WT mice, lead-treated MT-null mice showed dose-related nephromegaly. In addition, after lead exposure renal function was significantly diminished in MT-null mice in comparison to WT mice. MT-null mice accumulated less renal lead than WT mice and did not form lead inclusion bodies, which were present in the kidneys of WT mice. In gene array analysis, renal glutathione S-transferases were up-regulated after lead in MT-null mice only. In vitro studies on fibroblast cell lines derived from MT-null and WT mice showed that MT-null cells were much more sensitive to lead cytotoxicity. MT-null cells accumulated less lead and formed no inclusion bodies. The MT-null phenotype seems to preclude lead-induced inclusion body formation and increases lead toxicity at the organ and cellular level despite reducing lead accumulation. This study reveals important roles for MT in chronic lead toxicity, lead accumulation, and inclusion body formation. PMID- 11891202 TI - Mice lacking Smad3 are protected against cutaneous injury induced by ionizing radiation. AB - Transforming growth factor-beta (TGF-beta) plays a central role in the pathogenesis of inflammatory and fibrotic diseases, including radiation-induced fibrosis. We previously reported that mice null for Smad3, a key downstream mediator of TGF-beta, show accelerated healing of cutaneous incisional wounds with reduced inflammation and accumulation of matrix. To determine if loss of Smad3 decreases radiation-induced injury, skin of Smad3+/+ [wild-type (WT)] and /- [knockout (KO)] mice was exposed to a single dose of 30 to 50 Gy of gamma irradiation. Six weeks later, skin from KO mice showed significantly less epidermal acanthosis and dermal influx of mast cells, macrophages, and neutrophils than skin from WT littermates. Skin from irradiated KO mice exhibited less immunoreactive TGF-beta and fewer myofibroblasts, suggesting that these mice will have a significantly reduced fibrotic response. Although irradiation induced no change in the immunohistochemical expression of the TGF-beta type I receptor, the epidermal expression of the type II receptor was lost after irradiation whereas its dermal expression remained high. Primary keratinocytes and dermal fibroblasts prepared from WT and KO mice showed similar survival when irradiated, as did mice exposed to whole-body irradiation. These results suggest that inhibition of Smad3 might decrease tissue damage and reduce fibrosis after exposure to ionizing irradiation. PMID- 11891203 TI - Role of galectin-3 in breast cancer metastasis: involvement of nitric oxide. AB - We investigated the role of galectin-3 in metastasis of human breast carcinoma BT549 cells using the experimental liver metastasis model. Underlying mechanisms were then elucidated using the liver/tumor co-culture and cell culture systems. After intrasplenic injection, galectin-3 cDNA transfected BT549 cells (BT549(gal 3 wt)) formed metastatic colonies in the liver, while galectin-3 null BT549 cells (BT549(par)) did not, demonstrating that galectin-3 enhances metastatic potential. More than 90% of BT549(gal-3 wt) cells survived after 24 hours-co culture with the liver fragments isolated following ischemia treatment. In contrast, more than half of BT549(par) cells showed metabolic death following co culture with the liver fragments. When the liver from inducible nitric oxide synthase (iNOS) knockout mice was used, no cytotoxicity to BT549(par) cells was observed. Thus, iNOS exerts cytotoxicity on BT549(par) cells and galectin-3 can protect against iNOS-induced cytotoxicity. BT549(gal-3 wt) also exhibited enhanced survival against peroxynitrite (up to 400 micromol/L) in vitro. A single mutation in the NWGR motif of galectin-3 obliterated both metastatic capability and cell survival, indicating that the antiapoptotic function of galectin-3 is involved in enhanced metastasis. In conclusion, galectin-3 enhances the metastatic potential of BT549 cells through resistance to the products of iNOS, possibly through its bcl-2-like antiapoptotic function. PMID- 11891204 TI - Cold ischemia induces isograft arteriopathy, but does not augment allograft arteriopathy in non-immunosuppressed hosts. AB - Prolonged cold ischemia has been suggested as a factor that will exacerbate later graft arterial disease (GAD), a major limiting factor for long-term transplant survival. We therefore examined the effects of cold ischemia on GAD as well as adhesion molecule and cytokine expression in murine cardiac grafts. Mild GAD developed in isografts undergoing 4-hour cold ischemia. Relative to control isografts, cold ischemia induced transiently enhanced endothelial expression of intercellular adhesion molecule-1 (ICAM-1) at 4 hours post-transplant. There was also transiently-augmented gene expression of interleukin (IL)-1beta, IL-6, and transforming growth factor-beta in these cold-ischemic isografts. By 3 days post transplantation, however, there were no longer any differences between control and cold ischemic isografts. Cold ischemia did not significantly affect the final grade of either parenchymal rejection or GAD in long-term (4 to 12 weeks) major histocompatibility complex (MHC) I- or MHC II-mismatched allografts molecules transplanted without immunosuppression. At early time points after cold ischemia (4 to 24 hours), allografts mismatched for MHC I and/or MHC II showed enhanced expression of ICAM-1 and cytokines comparable to that seen in isografts. By day 7 post-transplant, both control and cold ischemia allografts showed comparable expression of cytokines and adhesion molecules. Although prolonged cold ischemia can initiate mild GAD in isografts by transiently enhancing antigen non-specific inflammatory responses, it does not significantly augment subsequent alloresponses. PMID- 11891205 TI - Doxycycline modulates smooth muscle cell growth, migration, and matrix remodeling after arterial injury. AB - The tetracyclines function as antibiotics by inhibiting bacterial protein synthesis, but recent work has shown that they are pluripotent drugs that affect many mammalian cell functions including proliferation, migration, apoptosis, and matrix remodeling. Because all of these processes have been implicated in arterial intimal lesion development, the objective of these studies was to examine the effect of doxycycline treatment using a well-characterized model of neointimal thickening, balloon catheter denudation of the rat carotid artery. Rats were treated with 30-mg/kg/day doxycycline. Doxycycline reduced the activity of matrix metalloproteinase (MMP)-2 and MMP-9 in the arterial wall, and inhibited smooth muscle cell migration from media to intima by 77% at 4 days after balloon injury. Replication of smooth muscle cells in the intima at 7 days was reduced from 28.3 plus minus 2.5% in controls to 17.0 +/- 2.8% in doxycycline-treated rats. The synthesis of elastin and collagen was not affected, but accumulation of elastin was blocked in the doxycycline-treated rats. By contrast, collagen accumulation was not affected, which led to the formation of a more collagen-rich intima. At 28 days after injury, the intimal:medial ratio was significantly reduced from 1.67 +/- 0.09 in control rats to 1.36 +/- 0.06 in the doxycycline treated rats. This study shows that doxycycline is an effective inhibitor of cell proliferation, migration, and MMP activity in vivo. Further study in more complicated models of atherosclerosis and restenosis is warranted. PMID- 11891206 TI - Inhibition of proliferative retinopathy by the anti-vascular agent combretastatin A4. AB - Retinal neovascularization occurs in a variety of diseases including diabetic retinopathy, the most common cause of blindness in the developed world. There is accordingly considerable incentive to develop drugs that target the aberrant angiogenesis associated with these conditions. Previous studies have shown that a number of anti-angiogenic agents can inhibit retinal neovascularization in a well characterized murine model of ischemia-induced proliferative retinopathy. Combretastatin-A4 (CA-4) is an anti-vascular tubulin-binding agent currently undergoing clinical evaluation for the treatment of solid tumors. We have recently shown that CA-4 is not tumor-specific but elicits anti-vascular effects in nonneoplastic angiogenic vessels. In this study we have examined the capacity of CA-4 to inhibit retinal neovascularization in vivo. CA-4 caused a dose dependent inhibition of neovascularization with no apparent side effects. The absence of vascular abnormalities or remnants of disrupted neovessels in retinas of CA-4-treated mice suggests an anti-angiogenic mechanism in this model, in contrast to the anti-vascular effects observed against established tumor vessels. Importantly, histological and immunohistochemical analyses indicated that CA-4 permitted the development of normal retinal vasculature while inhibiting aberrant neovascularization. These data are consistent with CA-4 eliciting tissue dependent anti-angiogenic effects and suggest that CA-4 has potential in the treatment of nonneoplastic diseases with an angiogenic component. PMID- 11891207 TI - Chromosomal imbalances in choroid plexus tumors. AB - We studied 49 choroid plexus tumors by comparative genomic hybridization. Chromosomal imbalances were found in 32 of 34 choroid plexus papillomas and 15 of 15 choroid plexus carcinomas. Choroid plexus papillomas frequently showed +7q (65%), +5q (62%), +7p (59%), +5p (56%), +9p (50%), +9q (41%), +12p, +12q (38%), and +8q (35%) as well as -10q (56%), -10p, and -22q (47%); choroid plexus carcinomas mainly showed +12p, +12q, +20p (60%), +1, +4q, +20q (53%), +4p (47%), +8q, +14q (40%), +7q, +9p, +21 (33%) as well as -22q (73%), -5q (40%), -5p, and 18q (33%). Several chromosomal imbalance differences could be found that were characteristic for a tumor entity or age group. In choroid plexus papillomas +5q, +6q, +7q, +9q, +15q, +18q, and -21q were significantly more common whereas choroid plexus carcinomas were characterized by +1, +4q, +10, +14q, +20q, +21q, 5q, -9p, -11, -15q, and -18q. Among choroid plexus papillomas, children more often showed +8q, +14q, +12, and +20q; adults mainly presented with +5q, +6q, +15q, +18q, and -22q. Although the number of aberrations overall as well as of gains and losses on their own bore no significance on survival among choroid plexus tumors, a significantly longer survival among patients with choroid plexus carcinomas was associated with +9p and -10q. Our results show that aberrations differ between choroid plexus papillomas and choroid plexus carcinomas as well as between pediatric and adult choroid plexus papillomas, supporting the notion of different genetic pathways. Furthermore, gain of 9p and loss of 10q seem to be correlated with a more favorable prognosis in choroid plexus carcinomas. PMID- 11891208 TI - Induction of tolerance to naphthalene in Clara cells is dependent on a stable phenotypic adaptation favoring maintenance of the glutathione pool. AB - Repeated exposures to the Clara cell cytotoxicant naphthalene (NA) result in target cell populations that become refractory to further injury. To determine whether tolerance occurs from specific adaptations favoring glutathione (GSH) resynthesis without broad shifts in cellular phenotype, mice were administered NA for 21 days. We found that gamma-glutamylcysteine synthetase (gamma-GCS) was induced in tolerant Clara cells by repeated exposures to NA. Treating tolerant mice with buthionine sulfoximine, a gamma-GCS inhibitor, eliminates resistance acquired by repeated exposures to NA. Broad phenotypic shifts were not present. Marker proteins of differentiation declined over the first 3 days in the development of tolerance, but returned to control levels at 14 and 21 days. Epithelial organizational structure and internal organelle composition in Clara cells from tolerant mice were similar compared to corn oil-treated controls, while subtle shifts in organelle distribution were present. We conclude that induction of gamma-GCS expression is coordinated with the development of NA tolerance, but induction of NA tolerance does not markedly alter Clara cell differentiation, epithelial organization, or organelle composition in bronchiolar epithelium. PMID- 11891209 TI - Frequent co-localization of Cox-2 and laminin-5 gamma2 chain at the invasive front of early-stage lung adenocarcinomas. AB - Laminin-5 is an extracellular matrix protein that plays a key role in cell migration and tumor invasion. Cox-2 is an induced isoform of cyclooxygenases that plays an important role in carcinogenesis, suppression of apoptosis, angiogenesis, and metastasis of colon cancer. We report frequent co-expression of cox-2 and laminin-5 at the invasive front of early-stage lung adenocarcinomas. We investigated the expression of cox-2 and laminin-5 immunohistochemically in 102 cases of small-sized lung adenocarcinoma (maximum dimension, 2 cm or less). Cox-2 and laminin-5 were expressed in 97 (95.1%) and 82 (80.4%) cases, respectively. Both were preferentially localized in cancer cells at the cancer-stroma interface, although cox-2 tended to show a diffuse staining pattern in some cases. A comparison of their staining patterns revealed a striking similarity in their distribution in 24 cases, and a partial overlap between their localization in another 20 cases. Moreover, an overall correlation was found between the expression levels of cox-2 and laminin-5 (P = 0.018). To gain insight into the mechanisms that regulate the expression of these proteins, we additionally studied their expression in 58 cases of stage I lung adenocarcinoma, in which p53 status was determined by immunohistochemistry, polymerase chain reaction-single strand conformation polymorphism analysis, and direct sequencing. The results showed that tumors with mutant p53 tended to express more cox-2 than those with wild-type p53 (P = 0.080). Also, tumors that overexpressed p53 had higher levels of cox-2 and laminin-5 than those without p53 overexpression (P = 0.032 and 0.047, respectively). Further immunohistochemical analysis showed that tumors that overexpressed both epidermal growth factor receptor (EGFR) and erbB-2 had higher levels of cox-2 and laminin-5 than those without concomitant overexpression of these proteins (P = 0.014 and P = 0.018, respectively). To see whether EGFR signaling is involved in cox-2 and laminin-5 expression, we further conducted in vitro analyses using six lung adenocarcinoma cell lines (A549, HLC 1, ABC-1, LC-2/ad, VMRC-LCD, and L27). Western blot analyses showed that cox-2 mRNA levels, and to a lesser extent laminin-5 gamma2 mRNA levels, correlated with the expression levels of erbB-2 and the phosphorylated form of MAPK/ERK-1/2 protein. The addition of transforming growth factor-alpha increased both cox-2 and laminin-5 gamma2 mRNA levels in A549, ABC-1, and L27 with different kinetics; the induction of cox-2 occurred earlier than that of laminin-5 gamma2. Finally, the migration of ABC-1 cells was inhibited by MAP kinase kinase inhibitor PD98059 and a selective cox-2 inhibitor NS-398. In contrast, the migration of A549 cells was inhibited by PD98059, but much less effectively by NS-398. These results suggest that co-stimulatory mechanisms may exist that increase the expression of cox-2 and laminin-5 at the invasive front of lung adenocarcinomas and that EGFR signaling could be one of the mechanisms. Further investigations are warranted concerning the role of cox-2 and laminin-5 in cancer cell invasion and the significance of p53 and EGFR signaling in the regulation of cox-2 and laminin-5 expression. PMID- 11891210 TI - Optical imaging of cancer metastasis to bone marrow: a mouse model of minimal residual disease. AB - The development of novel anti-cancer strategies requires more sensitive and less invasive methods to detect and monitor in vivo minimal residual disease in cancer models. Bone marrow metastases are indirectly detected by radiography as osteolytic and/or osteosclerotic lesions. Marrow micrometastases elude radiographic detection and, therefore, more sensitive methods are needed for their direct identification. Injection of cancer cells into the left cardiac ventricle of mice closely mimics micrometastatic spread. When luciferase transfected cells are used, whole-body bioluminescent reporter imaging can detect microscopic bone marrow metastases of approximately 0.5 mm(3) volume, a size below the limit in which tumors need to induce angiogenesis for further growth. This sensitivity translates into early detection of intramedullary tumor growth, preceding the appearance of a radiologically evident osteolysis by approximately 2 weeks. Bioluminescent reporter imaging also enables continuous monitoring in the same animal of growth kinetics for each metastatic site and guides end-point analyses specifically to the bones affected by metastatic growth. This model will accelerate the understanding of the molecular events in metastasis and the evaluation of novel therapies aiming at repressing initial stages of metastatic growth. PMID- 11891211 TI - Vascular immunotargeting of glucose oxidase to the endothelial antigens induces distinct forms of oxidant acute lung injury: targeting to thrombomodulin, but not to PECAM-1, causes pulmonary thrombosis and neutrophil transmigration. AB - Oxidative endothelial stress, leukocyte transmigration, and pulmonary thrombosis are important pathological factors in acute lung injury/acute respiratory distress syndrome (ALI/ARDS). Vascular immunotargeting of the H(2)O(2)-generating enzyme glucose oxidase (GOX) to the pulmonary endothelium causes an acute oxidative lung injury in mice.(1) In the present study we compared the pulmonary thrombosis and leukocyte transmigration caused by GOX targeting to the endothelial antigens platelet-endothelial cell adhesion molecule (PECAM) and thrombomodulin (TM). Both anti-PECAM and anti-TM delivered similar amounts of (125)I-GOX to the lungs and caused a dose-dependent, tissue-selective lung injury manifested within 2 to 4 hours by high lethality, vascular congestion, polymorphonuclear neutrophil (PMN) sequestration in the pulmonary vasculature, severe pulmonary edema, and tissue oxidation, yet at an equal dose, anti-TM/GOX inflicted more severe lung injury than anti-PECAM/GOX. Moreover, anti-TM/GOX induced injury was accompanied by PMN transmigration in the alveolar space, whereas anti-PECAM/GOX-induced injury was accompanied by PMN degranulation within vascular lumen without PMN transmigration, likely because of PECAM blockage. Anti TM/GOX caused markedly more severe pulmonary thrombosis than anti-PECAM/GOX, likely because of TM inhibition. These results indicate that blocking of specific endothelial antigens by GOX immunotargeting modulates important pathological features of the lung injury initiated by local generation of H(2)O(2) and that this approach provides specific and robust models of diverse variants of human ALI/ARDS in mice. In particular, anti-TM/GOX causes lung injury combining oxidative, prothrombotic, and inflammatory components characteristic of the complex pathological picture seen in human ALI/ARDS. PMID- 11891212 TI - Pituitary hyperplasia in glycoprotein hormone alpha subunit-, p18(INK4C)-, and p27(kip-1)-null mice: analysis of proteins influencing p27(kip-1) ubiquitin degradation. AB - Most spontaneously developing hyperplastic and neoplastic lesions of the pituitary occur in the anterior pituitary. Targeted disruption of various cell cycle proteins, including Rb, p27(kip1) (p27), and p18(INK4c) (p18), is associated with intermediate lobe pituitary hyperplasia. To develop a model of anterior pituitary proliferation to study the pathogenesis of pituitary tumors, we crossed the glycoprotein hormone alpha-subunit (alphaSU)-null mice that develop thyroid-stimulating hormone (TSH) cell hyperplasia with p18-null mice. The resulting offsprings developed accelerated enlargement of the anterior lobe with predominantly TSH cell hyperplasia. Immunohistochemical and histological analyses of these mice along with p27/p18 double-null mice, p18-null mice, and p27-null mice showed evidence of TSH, adrenocorticotropic hormone, prolactin, and luteinizing hormone hyperplasia. To determine whether there were alterations of p27 and the target proteins implicated in the ubiquitin degradation of p27 and other cyclin-dependent kinase inhibitors, we examined expression of SKP 2, Grb 2, and Jab 1 in the pituitaries of null mice. In the alphaSU-null mice there were decreased levels of SKP 2 and elevated levels of Grb 2 expression by Western blot analysis. Immunohistochemical analysis of the pituitary showed elevated Grb 2 in alphaSU-null and p18/alphaSU double-null mice. Jab 1 levels were not different from controls in the pituitary. These results show that 1) the p18/alphaSU double null mice represent a good model to study the rapid development of anterior pituitary hyperplasia, and 2) various proteins important in p27 and other cyclin dependent kinase inhibitor protein degradation are altered in the pituitary of alphaSU-null and p18/alphaSU double-null mouse models. PMID- 11891213 TI - Absence of decorin adversely influences tubulointerstitial fibrosis of the obstructed kidney by enhanced apoptosis and increased inflammatory reaction. AB - Decorin, a small dermatan-sulfate proteoglycan, participates in extracellular matrix assembly and influences directly and indirectly cell behavior via interactions with signaling membrane receptors and transforming growth factor (TGF)-beta. We have therefore compared the development of tubulointerstitial kidney fibrosis in wild-type (WT) and decorin-/- mice in the model of unilateral ureteral obstruction. Without obstruction, kidneys from decorin-/- mice did not differ in any aspect from their WT counterparts. However, already 12 hours after obstruction decorin-/- animals showed lower levels of p27(KIP1) and soon thereafter a more pronounced up-regulation and activation of initiator and effector caspases followed by enhanced apoptosis of tubular epithelial cells. Later, a higher increase of TGF-beta1 became apparent. After 7 days, there was an up to 15-fold transient up-regulation of the related proteoglycan biglycan, which was mainly caused by the appearance of biglycan-expressing mononuclear cells. Other small proteoglycans showed no similar response. Because of enhanced degradation of type I collagen, end-stage kidneys from decorin-/- animals were more atrophic than WT kidneys. These data suggest that decorin exerts beneficial effects on tubulointerstitial fibrosis, primarily by influencing the expression of a key cyclin-dependent kinase inhibitor and by limiting the degree of apoptosis, mononuclear cell infiltration, tubular atrophy, and expression of TGF beta1. PMID- 11891214 TI - Activation of extracellular signal-regulated kinase (ERK)1/2, but not p38 and c Jun N-terminal kinase, is involved in signaling of a novel cytokine, ML-1. AB - A novel cytokine, ML-1, was recently discovered, which shares a similar sequence homology with, but is functionally distinct from, IL-17 (Kawaguchi, M., Onuchic, L., Li, X. D., Essayan, D. M., Schroeder, J., Xiao, H. Q., Liu, M. C., Krishnaswamy, G., Germino, G., and Huang, S. K. (2001) J. Immunol. 167, 4430 4435). To determine the signaling mechanisms of ML-1, we investigated activation of mitogen-activated protein (MAP) kinases induced by ML-1. Results show that ML 1 induces in a time-dependent fashion the expression of IL-6 and IL-8 in both primary bronchial epithelial cells (PBECs) and human umbilical vein endothelial cells (HUVECs). ML-1 activated a MAP kinase and an extracellular signal-regulated kinase (ERK)1/2 but not p38 or the c-Jun N-terminal kinase (JNK) in both cell types. Selective MAP kinase kinase (MEK)1/2 inhibitors, PD98059 and U0126, inhibited, in a dose-dependent manner, ML-1-induced expression of IL-6 and IL-8. These findings suggest that ML-1-induced IL-6 and IL-8 production is mediated through the activation of ERK1/2 in both cell types. PMID- 11891215 TI - Aging and surface expression of hippocampal NMDA receptors. AB - Aging is known to alter many physiological processes within the brain including synaptic responses, long-term potentiation, learning, and memory. Aging has also been shown to alter the expression and distribution of N-methyl-d-aspartate (NMDA) receptors in many different brain regions, including the hippocampus. Additionally, we have recently reported that young adult rats show an activity dependent increase in the surface expression of NMDA receptors. We have extended these observations in the present study in aged animals and have found that aged Fischer 344 rats fail to show activity-dependent changes in the surface distribution of NMDA receptors. In conjunction with this observation we have also noted that aged rats show an expression deficit in the C2 splice variant of the NR1 subunit. This subunit is preferentially shifted to the surface following stimulation in young adult animals. As the NMDA receptor is thought to play an important role in neuronal signaling, these observations suggest possible new areas of dysfunction in this receptor that might underlie age-related deficits in neuronal physiology. PMID- 11891216 TI - The PDZ proteins PICK1, GRIP, and syntenin bind multiple glutamate receptor subtypes. Analysis of PDZ binding motifs. AB - Using sequence homology searches, yeast two-hybrid assays and glutathione S transferase (GST)-pull-down approaches we have identified a series of glutamate receptor subunits that interact differentially with the PDZ proteins GRIP, PICK1, and syntenin. GST-pull-down experiments identified more interactions than detected by yeast two-hybrid assays. We report several receptor-protein interactions, strong ones include: (i) GRIP and syntenin with mGluR7a, mGluR4a, and mGluR6; (ii) PICK1 and GRIP with mGluR3; and (iii) syntenin with all forms of GluR1-4 and mGluR7b. We further characterized the novel mGluR7a-GRIP interaction found both in yeast two-hybrid and GST-pull-down assays and observed that mGluR7a localization overlapped with GRIP with in hippocampal neurons. The wide range of targets for PICK1, GRIP, and syntenin suggests they may represent a molecular mechanism that can concentrate and/or regulate a number of different receptors at a common site on a synapse. These data also suggest that the structural determinants involved in PDZ interactions are more complex than originally envisaged. PMID- 11891217 TI - Allosteric effects potentiating the release of the second fibrinopeptide A from fibrinogen by thrombin. AB - Fibrin formation depends on the release of the two N-terminal fibrinopeptides A (FPA) from fibrinogen, and its formation is accompanied by an intermediate, alpha profibrin, which lacks only one of the FPA. In this study, we confirm that the maximal levels of alpha-profibrin found over the course of thrombin reactions with human fibrinogen are only half of what would be expected if the first and second FPA were being released independently with equal rate constants. The rapidity of release of the fibrinopeptides by thrombin had been shown to depend on an allosteric transformation that is induced when Na(+) binds to a site defined by the 215-227 residues of thrombin, a transformation that results in the exposure of its fibrinogen-binding exosites transforming the thrombin from a slow to a fast acting form toward fibrinogen. When choline was substituted for sodium to transform thrombin to its slow form, the maximal levels of alpha-profibrin rose to those expected for independent release of the two FPA. Thus, it is only the fast thrombin that releases the second FPA fast, and that fast release only occurs when both FPA are present because of a partial coupling of its release with that of the first FPA. The release of the FPA from purified alpha-profibrin with the first FPA already missing is no faster than the release of any FPA. Surprisingly, we also found that slow thrombin became increasingly transformed to a fast form in the absence of sodium when the fibrinogen was elevated to high concentrations. This potentiation by concentrated fibrinogen also occurs with the recombinant mutant thrombin (Y225P), which is otherwise slow in both the presence and absence of Na(+). The potentiation of thrombin by fibrinogen must be short lived so that the thrombin reverts to its slow acting form in the interim among encounters with other fibrinogen molecules in dilute fibrinogen solutions lacking Na(+), whereas at high fibrinogen concentrations the thrombin encounters other molecules before it reverts back to the slow form. PMID- 11891219 TI - A novel Src homology 2 domain-containing molecule, Src-like adapter protein-2 (SLAP-2), which negatively regulates T cell receptor signaling. AB - We have cloned a novel adapter protein containing Src homology 2 and Src homology 3 domains similar to the Src family of tyrosine kinases. This molecule lacks a catalytic tyrosine kinase domain and is related to a previously identified protein, Src-like adapter protein (SLAP), and is therefore designated SLAP-2. Northern blot analysis indicates that SLAP-2 is predominantly expressed in the immune system. Jurkat T cells express SLAP-2 protein and overexpression of SLAP-2 in these cells negatively regulates T cell receptor signaling as assessed by interleukin-2 promoter or NF-AT promoter reporter constructs. Mutational analysis revealed that an intact SH2 domain of SLAP-2 is essential for this inhibitory effect, whereas mutation of the SH3 domain alone has no effect. This inhibitory effect is upstream of the activation of Ras and increase of intracellular calcium levels, as no inhibition was observed when the cells were activated by phorbol ester plus ionomycin. SLAP-2 interacts with Cbl in vivo in a phosphorylation independent manner and with ZAP-70 and T cell receptor zeta chain upon T cell receptor activation. Finally, we show that the mutation of a predicted myristoylation site within the NH(2)-terminal of SLAP-2 is essential for its inhibitory effect. This report therefore implicates SLAP and SLAP-2 as a family of adapter proteins that negatively regulate T cell receptor signaling. PMID- 11891218 TI - alpha 2B-adrenergic receptor activates MAPK via a pathway involving arachidonic acid metabolism, matrix metalloproteinases, and epidermal growth factor receptor transactivation. AB - We have investigated the mechanisms whereby alpha(2B)-adrenergic receptor (alpha(2B)-AR) promotes MAPK activation in a clone of the renal tubular cell line, LLC-PK1, transfected with the rat nonglycosylated alpha(2)-AR gene. Treatment of LLC-PK1-alpha(2B) with UK14304 or dexmedetomidine caused arachidonic acid (AA) release and ERK2 phosphorylation. AA release was abolished by prior treatment of the cells with pertussis toxin, quinacrine, or methyl arachidonyl fluorophosphonate but not by the addition of the MEK inhibitor U0126. The effects of alpha(2)-agonists on MAPK phosphorylation were mimicked by cell exposure to exogenous AA. On the other hand, quinacrine abolished the effects of UK14304, but not of AA, suggesting that AA released through PLA2 is responsible for MAPK activation by alpha(2B)-AR. The effects of alpha(2)-agonists or AA were PKC independent and were attenuated by indomethacin and nordihydroguaiaretic acid. Treatment with batimastat, CRM 197, or tyrphostin AG1478 suppressed MAPK phosphorylation promoted by alpha(2)-agonist or AA. Furthermore, conditioned culture medium from UK14304-treated LLC-PK1-alpha(2B) induced MAPK phosphorylation in wild-type LLC-PK1. Based on these data, we propose a model whereby activation of MAPK by alpha(2B)-AR is mediated through stimulation of PLA2, AA release, generation of AA derivatives, activation of matrix metalloproteinases, release of heparin-binding EGF-like growth factor, transactivation of epidermal growth factor receptor, and recruitment of Shc. Whether this pathway is particular to alpha(2B)-AR and LLC-PK1 or whether it can be extended to other cell types and/or other G-protein-coupled receptors remains to be established. PMID- 11891220 TI - Chloroplast YidC homolog Albino3 can functionally complement the bacterial YidC depletion strain and promote membrane insertion of both bacterial and chloroplast thylakoid proteins. AB - A new component of the bacterial translocation machinery, YidC, has been identified that specializes in the integration of membrane proteins. YidC is homologous to the mitochondrial Oxa1p and the chloroplast Alb3, which functions in a novel pathway for the insertion of membrane proteins from the mitochondrial matrix and chloroplast stroma, respectively. We find that Alb3 can functionally complement the Escherichia coli YidC depletion strain and promote the membrane insertion of the M13 procoat and leader peptidase that were previously shown to depend on the bacterial YidC for membrane translocation. In addition, the chloroplast Alb3 that is expressed in bacteria is essential for the insertion of chloroplast cpSecE protein into the bacterial inner membrane. Surprisingly, Alb3 is not required for the insertion of cpSecE into the thylakoid membrane. These results underscore the importance of Oxa1p homologs for membrane protein insertion in bacteria and demonstrate that the requirement for Oxa1p homologs is different in the bacterial and thylakoid membrane systems. PMID- 11891222 TI - Elovl1 and p55Cdc genes are localized in a tail-to-tail array and are co expressed in proliferating cells. AB - Elovl1 is a ubiquitously expressed gene, the product of which belongs to a highly conserved family of microsomal enzymes, which are involved in the formation of very long chain fatty acids and sphingolipids in yeast to man. To elucidate the structure and regulation of the Elovl gene we have isolated a lambda phage genomic DNA clone containing the entire mouse gene and found that Elovl1 consists of eight exons that are dispersed over 5.4 kb of genomic sequence. Interestingly, sequencing of the lambda clone to completion revealed that the insert contained a segment of the cell cycle gene p55Cdc directed in the opposite orientation. The genes are very tightly linked so that the 3'-end of the long mRNA species are complementary over a short stretch of nucleotides. Although both Elovl1 and p55Cdc are highly conserved genes, a BLAST search implies that the tail-to-tail arrangement has evolved in vertebrates. Despite the non-similar expression pattern in different tissues, mRNA analysis of the two genes disclosed simultaneous transcription during a proliferation-differentiation transition state, which suggests that the two genes may be regulated through a common bi directional transcription mechanism under specific conditions. PMID- 11891221 TI - Identification by site-directed mutagenesis of residues involved in ligand recognition and activation of the human A3 adenosine receptor. AB - Ligand recognition has been extensively explored in G protein-coupled A(1), A(2A), and A(2B) adenosine receptors but not in the A(3) receptor, which is cerebroprotective and cardioprotective. We mutated several residues of the human A(3) adenosine receptor within transmembrane domains 3 and 6 and the second extracellular loop, which have been predicted by previous molecular modeling to be involved in the ligand recognition, including His(95), Trp(243), Leu(244), Ser(247), Asn(250), and Lys(152). The N250A mutant receptor lost the ability to bind both radiolabeled agonist and antagonist. The H95A mutation significantly reduced affinity of both agonists and antagonists. In contrast, the K152A (EL2), W243A (6.48), and W243F (6.48) mutations did not significantly affect the agonist binding but decreased antagonist affinity by approximately 3-38-fold, suggesting that these residues were critical for the high affinity of A(3) adenosine receptor antagonists. Activation of phospholipase C by wild type (WT) and mutant receptors was measured. The A(3) agonist 2-chloro-N(6)-(3-iodobenzyl)-5'-N methylcarbamoyladenosine stimulated phosphoinositide turnover in the WT but failed to evoke a response in cells expressing W243A and W243F mutant receptors, in which agonist binding was less sensitive to guanosine 5'-gamma thiotriphosphate than in WT. Thus, although not important for agonist binding, Trp(243) was critical for receptor activation. The results were interpreted using a rhodopsin-based model of ligand-A(3) receptor interactions. PMID- 11891223 TI - Transcriptional control of monolignol biosynthesis in Pinus taeda: factors affecting monolignol ratios and carbon allocation in phenylpropanoid metabolism. AB - Transcriptional profiling of the phenylpropanoid pathway in Pinus taeda cell suspension cultures was carried out using quantitative real time PCR analyses of all known genes involved in the biosynthesis of the two monolignols, p-coumaryl and coniferyl alcohols (lignin/lignan precursors). When the cells were transferred to a medium containing 8% sucrose and 20 mm potassium iodide, the monolignol/phenylpropanoid pathway was induced, and transcript levels for phenylalanine ammonia lyase, cinnamate 4-hydroxylase, p-coumarate 3-hydroxylase, 4-coumarate:CoA ligase, caffeoyl-CoA O-methyltransferase, cinnamoyl-CoA reductase, and cinnamyl alcohol dehydrogenase were coordinately up-regulated. Provision of increasing levels of exogenously supplied Phe to saturating levels (40 mm) to the induction medium resulted in further up-regulation of their transcript levels in the P. taeda cell cultures; this in turn was accompanied by considerable increases in both p-coumaryl and coniferyl alcohol formation and excretion. By contrast, transcript levels for both cinnamate 4-hydroxylase and p coumarate 3-hydroxylase were only slightly up-regulated. These data, when considered together with metabolic profiling results and genetic manipulation of various plant species, reveal that carbon allocation to the pathway and its differential distribution into the two monolignols is controlled by Phe supply and differential modulation of cinnamate 4-hydroxylase and p-coumarate 3 hydroxylase activities, respectively. The coordinated up-regulation of phenylalanine ammonia lyase, 4-coumarate:CoA ligase, caffeoyl-CoA O methyltransferase, cinnamoyl-CoA reductase and cinnamyl alcohol dehydrogenase in the presence of increasing concentrations of Phe also indicates that these steps are not truly rate-limiting, because they are modulated according to metabolic demand. Finally, the transcript profile of a putative acid/ester O methyltransferase, proposed as an alternative catalyst for O-methylation leading to coniferyl alcohol, was not up-regulated under any of the conditions employed, suggesting that it is not, in fact, involved in monolignol biosynthesis. PMID- 11891224 TI - Cross-repression, a functional consequence of the physical interaction of non liganded nuclear receptors and POU domain transcription factors. AB - Nuclear receptors (NRs) and POU domain factors form two important transcription factor families for which several levels of functional interference have been described. In this study, the adopted orphan receptors constitutive androstane receptor (CAR) and pregnane X receptor (PXR) were found to perform direct protein protein interactions with Pit-1, a representative POU domain factor. The ligand dependent interaction profile of Pit-1 with CAR, PXR, and the vitamin D receptor in solution was shown to be that of a corepressor. In the absence of receptor agonist Pit-1 inhibited the complex formation of NRs with the retinoid X receptor on DNA. Also in living cells, Pit-1 and Oct-1, another POU domain factor, behaved like corepressors of NR signaling, and Pit-1-mediated repression was found to involve histone deacetylases. Conversely vitamin D receptor, CAR, and PXR were shown to act as repressors of Pit-1 signaling in different cell lines (MCF-7, HaCaT, and GH4C1). This repression was found to be independent of histone deacetylases and seems to be based on a competition of NRs with coactivator and corepressor proteins for overlaying interaction interfaces on the surface of Pit 1. Taken together this study suggests that cross-repression should occur in all tissues in which POU domain factors and non-liganded NRs meet each other. PMID- 11891225 TI - Laminin-10/11 and fibronectin differentially prevent apoptosis induced by serum removal via phosphatidylinositol 3-kinase/Akt- and MEK1/ERK-dependent pathways. AB - Cell adhesion to the extracellular matrix inhibits apoptosis, but the molecular mechanisms underlying the signals transduced by different matrix components are not well understood. Here, we examined integrin-mediated antiapoptotic signals from laminin-10/11 in comparison with those from fibronectin, the best characterized extracellular adhesive ligand. We found that the activation of protein kinase B/Akt in cells adhering to laminin-10/11 can rescue cell apoptosis induced by serum removal. Consistent with this, wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase, or ectopic expression of a dominant-negative mutant of Akt selectively accelerated cell death upon serum removal. In contrast to laminin-10/11, fibronectin rescued cells from serum depletion-induced apoptosis mainly through the extracellular signal-regulated kinase pathway. Cell survival on fibronectin but not laminin was significantly reduced by treatment with PD98059, a specific inhibitor of mitogen- or extracellular signal-regulated kinase kinase-1 (MEK1) and by expression of a dominant-negative mutant of MEK1. Laminin-10/11 was more potent than fibronectin in preventing apoptosis induced by serum depletion. These results, taken together, demonstrate laminin-10/11 potency as a survival factor and demonstrate that different extracellular matrix components can transduce distinct survival signals through preferential activation of subsets of multiple integrin-mediated signaling pathways. PMID- 11891226 TI - Egg shell collagen formation in Caenorhabditis elegans involves a novel prolyl 4 hydroxylase expressed in spermatheca and embryos and possessing many unique properties. AB - The collagen prolyl 4-hydroxylases (EC ) play a critical role in the synthesis of all collagens. The enzymes from all vertebrate species studied are alpha(2)beta(2) tetramers, in which the beta subunit is identical to protein disulfide isomerase (PDI). Two isoforms of the catalytic alpha subunit, PHY-1 and PHY-2, have previously been characterized from Caenorhabditis elegans. We report here on the cloning and characterization of a third C. elegans alpha subunit isoform, PHY-3. It is much shorter than the previously characterized vertebrate and C. elegans alpha subunits and shows 23-30% amino acid sequence identity to PHY-1 and PHY-2 within the catalytic C-terminal region. Recombinant PHY-3 coexpressed in insect cells with a C. elegans PDI isoform that does not associate with PHY-1 was found to be an active prolyl 4-hydroxylase. The phy-3 gene consists of five exons, and its expression pattern differs distinctly from the hypodermally expressed phy-1 and phy-2 in that it is expressed in embryos, late larval stages, and adult nematodes, expression in the latter being restricted to the spermatheca. Nematodes homozygous for a phy-3 deletion are phenotypically of the wild type and fertile, but the 4-hydroxyproline content of phy-3(-/-) early embryos was reduced by about 90%. PHY-3 is thus likely to be involved in the synthesis of collagens in early embryos, probably of those in the egg shell. PMID- 11891227 TI - Functionality of alternative splice forms of the first enzymes involved in human molybdenum cofactor biosynthesis. AB - In humans, genetic deficiencies of enzymes involved in molybdenum cofactor biosynthesis trigger an autosomal recessive and usually fatal disease with severe mostly neurological symptoms. In each of the three biosynthesis steps, at least two proteins or domains are linked for catalysis. For steps 1 and 2, bicistronic mocs (molybdenum cofactor synthesis) mRNAs were found (mocs1 and mocs2) that have been proposed to encode two separate proteins (A and B). In both cases, the A proteins share a highly conserved ubiquitin-like double glycine motif, which is functionally important at least for the small subunit of molybdopterin (MPT) synthase (MOCS2A). Besides the bicistronic form of mocs1, two alternative splice transcripts were found, resulting in the expression of multidomain proteins embodying both MOCS1A, but without the double glycine motif, and the entire MOCS1B. Here we describe the first functional characterization of the human proteins MOCS1A and MOCS1B as well as the MOCS1A-MOCS1B fusion proteins that catalyze the formation of precursor Z, a 6-alkyl pterin with a cyclic phosphate, the immediate precursor of MPT in molybdenum cofactor biosynthesis. High level expression of MOCS1A and MOCS1B in Escherichia coli resulted in the formation and accumulation of precursor Z that was subsequently converted to MPT. We showed that for catalytic activity MOCS1A needs an accessible C-terminal double glycine motif. In the MOCS1A-MOCS1B fusion proteins lacking the MOCS1A double glycines, only MOCS1B activity could be detected. No evidence was found for an expression of MOCS1B from the bicistronic mocs1A-mocs1B splice type I cDNA, indicating that MOCS1B is only expressed as a fusion to an inactive MOCS1A. Comparative mutational studies of MOCS1A and the small subunit of the E. coli MPT synthase (MoaD) indicate a different function of the double glycine motifs in both proteins. PMID- 11891228 TI - c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury. Inhibition by a novel DNA enzyme targeting c-Jun. AB - Neointima formation is a characteristic feature of common vascular pathologies, such as atherosclerosis and post-angioplasty restenosis, and involves smooth muscle cell proliferation. Determination of whether the bZIP transcription factor c-Jun plays a direct regulatory role in arterial lesion formation, or indeed in other disease, has been hampered by the lack of a potent and specific pharmacological inhibitor. c-Jun is poorly expressed in the uninjured artery wall and transiently induced following arterial injury in animal models. Here we generated a gene-specific DNAzyme-targeting c-Jun. We show that c-Jun protein is expressed in human atherosclerotic lesions. Dz13, a catalytically active c-Jun DNAzyme, cleaved c-Jun RNA and inhibited inducible c-Jun protein expression in vascular smooth muscle cells. Dz13 blocked vascular smooth muscle cell proliferation with potency exceeding its exact non-catalytic antisense oligodeoxynucleotide equivalent. Moreover, Dz13 abrogated smooth muscle cell repair following scraping injury in vitro and intimal thickening in injured rat carotid arteries in vivo. These studies demonstrate the positive influence on neointima formation by c-Jun and the therapeutic potential of a DNAzyme controlling its expression. PMID- 11891230 TI - Mannose-binding lectin (MBL) mutants are susceptible to matrix metalloproteinase proteolysis: potential role in human MBL deficiency. AB - Mannose-binding lectin (MBL) plays a critical role in innate immunity. Point mutations in the collagen-like domain (R32C, G34D, or G37E) of MBL cause a serum deficiency, predisposing patients to infections and diseases such as rheumatoid arthritis. We examined whether MBL mutants show enhanced susceptibility to proteolysis by matrix metalloproteinases (MMPs), which are important mediators in inflammatory tissue destruction. Human and rat MBL were resistant to proteolysis in the native state but were cleaved selectively within the collagen-like domain by multiple MMPs after heat denaturation. In contrast, rat MBL with mutations homologous to those of the human variants (R23C, G25D, or G28E) was cleaved efficiently without denaturation in the collagen-like domain by MMP-2 and MMP-9 (gelatinases A and B) and MMP-14 (membrane type-1 MMP), as well as by MMP-1 (collagenase-1), MMP-8 (neutrophil collagenase), MMP-3 (stromelysin-1), neutrophil elastase, and bacterial collagenase. Sites and order of cleavage of the rat MBL mutants for MMP-2 and MMP-9 were: Gly(45)-Lys(46) --> Gly(51)-Ser(52) --> Gly(63)-Gln(64) --> Asn(80)-Met(81) which differed from that of MMP-14, Gly(39)-Leu(40) --> Asn(80)-Met(81), revealing that the MMPs were not functionally interchangeable. These sites were homologous to those cleaved in denatured human MBL. Hence, perturbation of the collagen-like structure of MBL by natural mutations or by denaturation renders MBL susceptible to MMP cleavage. MMPs are likely to contribute to MBL deficiency in individuals with variant alleles and may also be involved in clearance of MBL and modulation of the host response in normal individuals. PMID- 11891229 TI - Biosynthesis of HNK-1 glycans on O-linked oligosaccharides attached to the neural cell adhesion molecule (NCAM): the requirement for core 2 beta 1,6-N acetylglucosaminyltransferase and the muscle-specific domain in NCAM. AB - The HNK-1 glycan, sulfo-->3GlcAbeta1-->3Galbeta1-->4GlcNAcbeta1-->R, is highly expressed in neuronal cells and apparently plays critical roles in neuronal cell migration and axonal extension. The HNK-1 glycan synthesis is initiated by the addition of beta1,3-linked GlcA to N-acetyllactosamine followed by sulfation of the C-3 position of GlcA. The cDNAs encoding beta1,3-glucuronyltransferase (GlcAT P) and HNK-1 sulfotransferase (HNK-1ST) have been recently cloned. Among various adhesion molecules, the neural cell adhesion molecule (NCAM) was shown to contain HNK-1 glycan on N-glycans. In the present study, we first demonstrated that NCAM also bears HNK-1 glycan attached to O-glycans when NCAM contains the O-glycan attachment scaffold, muscle-specific domain, and is synthesized in the presence of core 2 beta1,6-N-acetylglucosaminyltransferase, GlcAT-P, and HNK-1ST. Structural analysis of the HNK-1 glycan revealed that the HNK-1 glycan is attached on core 2 branched O-glycans, sulfo-->3GlcAbeta1-->3Galbeta1- >4GlcNAcbeta1-->6(Galbeta1-->3)GalNAc. Using synthetic oligosaccharides as acceptors, we found that GlcAT-P and HNK-1ST almost equally act on oligosaccharides, mimicking N- and O-glycans. By contrast, HNK-1 glycan was much more efficiently added to N-glycans than O-glycans when NCAM was used as an acceptor. These results are consistent with our results showing that HNK-1 glycan is minimally attached to O-glycans of NCAM in fetal brain, heart, and the myoblast cell line, C2C12. These results combined together indicate that HNK-1 glycan can be synthesized on core 2 branched O-glycans but that the HNK-1 glycan is preferentially added on N-glycans over O-glycans of NCAM, probably because N glycans are extended further than O-glycans attached to NCAM containing the muscle-specific domain. PMID- 11891231 TI - Cloning and characterization of the human factor XI gene promoter: transcription factor hepatocyte nuclear factor 4alpha (HNF-4alpha ) is required for hepatocyte specific expression of factor XI. AB - Factor XI is the zymogen of a plasma protease produced primarily in liver that is required for normal blood coagulation. We cloned approximately 2600 base pairs of the human factor XI gene upstream of exon one, identified transcription start sites, and conducted a functional analysis. Luciferase reporter assays demonstrate that the 381 base pairs upstream of exon one are sufficient for maximum promoter activity in HepG2 hepatocellular carcinoma cells. The removal of 19 base pairs between -381 and -363 results in a nearly complete loss of promoter activity. This region contains the sequence ACTTTG, a motif required for binding of the transcription factor hepatocyte nuclear factor 4alpha (HNF-4alpha) to the promoters of several genes. Gel mobility shift assays using HepG2 or rat hepatocyte nuclear extract confirm HNF-4alpha binds between bp -375 and -360. Scrambling the ACTTTG motif completely abolishes promoter activity in luciferase assays. The factor XI promoter functions poorly when transfected into HeLa carcinoma cells, and gel mobility shift experiments with HeLa nuclear extracts demonstrate no HNF-4alpha binding to the ACTTTG sequence. When a rat HNF-4alpha expression construct is co-transfected into HeLa cells, factor XI promoter activity is enhanced approximately 10-fold. We conclude that HNF-4alpha is required for hepatocyte-specific expression of factor XI. PMID- 11891233 TI - A long and steep ladder to the top. PMID- 11891232 TI - Aquaporin deletion in mice reduces corneal water permeability and delays restoration of transparency after swelling. AB - Two aquaporin (AQP)-type water channels are expressed in mammalian cornea, AQP1 in endothelial cells and AQP5 in epithelial cells. To test whether these aquaporins are involved in corneal fluid transport and transparency, we compared corneal thickness, water permeability, and response to experimental swelling in wild type mice and transgenic null mice lacking AQP1 and AQP5. Corneal thickness in fixed sections was remarkably reduced in AQP1 null mice and increased in AQP5 null mice. By z-scanning confocal microscopy, corneal thickness in vivo was (in microm, mean +/- S.E., n = 5 mice) 123 +/- 1 (wild type), 101 +/- 2 (AQP1 null), and 144 +/- 2 (AQP5 null). After exposure of the external corneal surface to hypotonic saline (100 mosm), the rate of corneal swelling (5.0 +/- 0.3 microm/min, wild type) was reduced by AQP5 deletion (2.7 +/- 0.1 microm/min). After exposure of the endothelial surface to hypotonic saline by anterior chamber perfusion, the rate of corneal swelling (7.1 +/- 1.0 microm/min, wild type) was reduced by AQP1 deletion (1.6 +/- 0.4 microm/min). Base-line corneal transparency was not impaired by AQP1 or AQP5 deletion. However, the recovery of corneal transparency and thickness after hypotonic swelling (10-min exposure of corneal surface to hypotonic saline) was remarkably delayed in AQP1 null mice with approximately 75% recovery at 7 min in wild type mice compared with 5% recovery in AQP1 null mice. Our data indicate that AQP1 and AQP5 provide the principal routes for corneal water transport across the endothelial and epithelial barriers, respectively. The impaired recovery of corneal transparency in AQP1 null mice provides evidence for the involvement of AQP1 in active extrusion of fluid from the corneal stroma across the corneal endothelium. The up-regulation of AQP1 expression and/or function in corneal endothelium may reduce corneal swelling and opacification following injury. PMID- 11891234 TI - An accidental plant biologist. PMID- 11891235 TI - Nitric oxide and abscisic acid cross talk in guard cells. PMID- 11891236 TI - Purine biosynthesis. Big in cell division, even bigger in nitrogen assimilation. PMID- 11891237 TI - Gene-containing regions of wheat and the other grass genomes. AB - Deletion line-based high-density physical maps revealed that the wheat (Triticum aestivum) genome is partitioned into gene-rich and -poor compartments. Available deletion lines have bracketed the gene-containing regions to about 10% of the genome. Emerging sequence data suggest that these may further be partitioned into "mini" gene-rich and gene-poor regions. An average of about 10% of each gene-rich region seem to contain genes. Sequence analyses in various species suggest that uneven distribution of genes may be a characteristic of all grasses and perhaps all higher organisms. Comparison of the physical maps with genetic linkage maps showed that recombination in wheat and barley (Hordeum vulgare) is confined to the gene-containing regions. Number of genes, gene density, and the extent of recombination vary greatly among the gene-rich regions. The gene order, relative region size, and recombination are highly conserved within the tribe Triticeae and moderately conserved within the family. Gene-poor regions are composed of retrotransposon-like non-transcribing repeats and pseudogenes. Direct comparisons of orthologous regions indicated that gene density in wheat is about one-half compared with rice (Oryza sativa). Genome size difference between wheat and rice is, therefore, mainly because of amplification of the gene-poor regions. Presence of species-, genera-, and family-specific repeats reveal a repeated invasion of the genomes by different retrotransposons over time. Preferential transposition to adjacent locations and presence of vital genes flanking a gene-rich region may have restricted retrotransposon amplification to gene-poor regions, resulting into tandem blocks of non-transcribing repeats. Insertional inactivation by adjoining retro-elements and selection seem to have played a major role in stabilizing genomes. PMID- 11891238 TI - Adenosine kinase deficiency is associated with developmental abnormalities and reduced transmethylation. AB - Adenosine (Ado) kinase (ADK; ATP:Ado 5' phosphotransferase, EC 2.7.1.20) catalyzes the salvage synthesis of adenine monophosphate from Ado and ATP. In Arabidopsis, ADK is encoded by two cDNAs that share 89% nucleotide identity and are constitutively, yet differentially, expressed in leaves, stems, roots, and flowers. To investigate the role of ADK in plant metabolism, lines deficient in this enzyme activity have been created by sense and antisense expression of the ADK1 cDNA. The levels of ADK activity in these lines range from 7% to 70% of the activity found in wild-type Arabidopsis. Transgenic plants with 50% or more of the wild-type activity have a normal morphology. In contrast, plants with less than 10% ADK activity are small with rounded, wavy leaves and a compact, bushy appearance. Because of the lack of elongation of the primary shoot, the siliques extend in a cluster from the rosette. Fertility is decreased because the stamen filaments do not elongate normally; hypocotyl and root elongation are reduced also. The hydrolysis of S-adenosyl-L-homo-cysteine (SAH) produced from S-adenosyl L-methionine (SAM)-dependent methylation reactions is a key source of Ado in plants. The lack of Ado salvage in the ADK-deficient lines leads to an increase in the SAH level and results in the inhibition of SAM-dependent transmethylation. There is a direct correlation between ADK activity and the level of methylesterified pectin in seed mucilage, as monitored by staining with ruthenium red, immunofluorescence labeling, or direct assay. These results indicate that Ado must be steadily removed by ADK to prevent feedback inhibition of SAH hydrolase and maintain SAM utilization and recycling. PMID- 11891240 TI - Interaction of the Arabidopsis E2F and DP proteins confers their concomitant nuclear translocation and transactivation. AB - E2F transcription factors are required for the progression and arrest of the cell cycle in animals. Like animals, plants have evolved to conserve the E2F family. The Arabidopsis genome encodes E2F and DP proteins that share a high similarity with the animal E2F and DP families. Here, we show that Arabidopsis E2F and DP proteins are not predominantly localized to the nucleus in analyses with green fluorescent protein, and that the complete nuclear localization of some members is driven by the co-expression of their specific partner proteins. Both AtE2F1 and AtE2F3 were translocated to the nucleus and transactivate an E2F reporter gene when co-expressed with DPa but not DPb. In contrast, AtE2F2 was inactive for both nuclear translocation and transactivation even when Dpa or DPb was co expressed. Because the DNA binding activities of the three E2Fs are equally stimulated by the interaction with DPa or DPb in vitro, the observed transactivation of AtE2F1 and AtE2F3 is DPa specific and nuclear import dependent. A green fluorescent protein fusion with an AtE2F3 mutant, in which a conserved nuclear export signal-like sequence in the dimerization domain was deleted, was localized to the nucleus. Thus, the concomitant nuclear translocation seems to be conferred by the DPa interaction to release an activity that inhibits an intrinsic nuclear import activity of AtE2Fs. Furthermore, the nuclear translocation of AtE2F3 stimulated by DPa was abolished by the deletion of the N-terminal region of AtE2F3, which is conserved among all the E2F proteins identified in plants to date. Replacement of the N-terminal region of AtE2F3 with a canonical nuclear localization signal only partially mimicked the effect of the DPa co-expression, demonstrating the function of plant E2F distinct from that observed for animal E2Fs. These observations suggest that the function of plant E2F and DP proteins is primarily controlled by their nuclear localization mediated by the interaction with specific partner proteins. PMID- 11891239 TI - Temperature-induced extended helix/random coil transitions in a group 1 late embryogenesis-abundant protein from soybean. AB - Group 1 late embryogenesis-abundant (LEA) proteins are a subset of hydrophilins that are postulated to play important roles in protecting plant macromolecules from damage during freezing, desiccation, or osmotic stress. To better understand the putative functional roles of group 1 LEA proteins, we analyzed the structure of a group 1 LEA protein from soybean (Glycine max). Differential scanning calorimetry of the purified, recombinant protein demonstrated that the protein assumed a largely unstructured state in solution. In the presence of trifluoroethanol (50% [w/v]), the protein acquired a 30% alpha-helical content, indicating that the polypeptide is highly restricted to adopt alpha-helical structures. In the presence of sodium dodecyl sulfate (1% [w/v]), 8% of the polypeptide chain adopted an alpha-helical structure. However, incubation with phospholipids showed no effect on the protein structure. Ultraviolet absorption and circular dichroism spectroscopy revealed that the protein existed in equilibrium between two conformational states. Ultraviolet absorption spectroscopy studies also showed that the protein became more hydrated upon heating. Furthermore, circular dichroism spectral measurements indicated that a minimum of 14% of amino acid residues existed in a solvent-exposed, left-handed extended helical or poly (L-proline)-type (PII) conformation at 20 degrees C with the remainder of the protein being unstructured. The content of PII-like structure increased as temperature was lowered. We hypothesize that by favoring the adoption of PII structure, instead of the formation of alpha-helical or beta sheet structures, group 1 LEA proteins retain a high content of surface area available for interaction with the solvent. This feature could constitute the basis of a potential role of LEA proteins in preventing freezing, desiccation, or osmotic stress damage. PMID- 11891241 TI - Simultaneous suppression of multiple genes by single transgenes. Down-regulation of three unrelated lignin biosynthetic genes in tobacco. AB - Many reports now describe the manipulation of plant metabolism by suppressing the expression of single genes. The potential of such work could be greatly expanded if multiple genes could be coordinately suppressed. In the work presented here, we test a novel method for achieving this by using single chimeric constructs incorporating partial sense sequences for multiple genes to target suppression of two or three lignin biosynthetic enzymes. We compare this method with a more conventional approach to achieving the same end by crossing plants harboring different antisense transgenes. Our results indicate that crossing antisense plants is less straightforward and predictable in outcome than anticipated. Most progeny had higher levels of target enzyme activity than predicted and had lost the expected modifications to lignin structure. In comparison, plants transformed with the chimeric partial sense constructs had more consistent high level suppression of target enzymes and had significant changes to lignin content, structure, and composition. It was possible to suppress three target genes coordinately using a single chimeric construct. Our results indicate that chimeric silencing constructs offer great potential for the rapid and coordinate suppression of multiple genes on diverse biochemical pathways and that the technique therefore deserves to be adopted by other researchers. PMID- 11891242 TI - Plant expansins are a complex multigene family with an ancient evolutionary origin. AB - Expansins are a group of extracellular proteins that directly modify the mechanical properties of plant cell walls, leading to turgor-driven cell extension. Within the completely sequenced Arabidopsis genome, we identified 38 expansin sequences that fall into three discrete subfamilies. Based on phylogenetic analysis and shared intron patterns, we propose a new, systematic nomenclature of Arabidopsis expansins. Further phylogenetic analysis, including expansin sequences found here in monocots, pine (Pinus radiata, Pinus taeda), fern (Regnellidium diphyllum, Marsilea quadrifolia), and moss (Physcomitrella patens) indicate that the three plant expansin subfamilies arose and began diversifying very early in, if not before, colonization of land by plants. Closely related "expansin-like" sequences were also identified in the social amoeba, Dictyostelium discoidium, suggesting that these wall-modifying proteins have a very deep evolutionary origin. PMID- 11891244 TI - Evidence supporting a role of jasmonic acid in Arabidopsis leaf senescence. AB - In this work, the role of jasmonic acid (JA) in leaf senescence is examined. Exogenous application of JA caused premature senescence in attached and detached leaves in wild-type Arabidopsis but failed to induce precocious senescence of JA insensitive mutant coi1 plants, suggesting that the JA-signaling pathway is required for JA to promote leaf senescence. JA levels in senescing leaves are 4 fold higher than in non-senescing ones. Concurrent with the increase in JA level in senescing leaves, genes encoding the enzymes that catalyze most of the reactions of the JA biosynthetic pathway are differentially activated during leaf senescence in Arabidopsis, except for allene oxide synthase, which is constitutively and highly expressed throughout leaf development. Arabidopsis lipoxygenase 1 (cytoplasmic) expression is greatly increased but lipoxygenase 2 (plastidial) expression is sharply reduced during leaf senescence. Similarly, AOC1 (allene oxide cyclase 1), AOC2, and AOC3 are all up-regulated, whereas AOC4 is down-regulated with the progression of leaf senescence. The transcript levels of 12-oxo-PDA reductase 1 and 12-oxo-PDA reductase 3 also increase in senescing leaves, as does PED1 (encoding a 3-keto-acyl-thiolase for beta-oxidation). This represents the first report, to our knowledge, of an increase in JA levels and expression of oxylipin genes during leaf senescence, and indicates that JA may play a role in the senescence program. PMID- 11891243 TI - Copper amine oxidase expression in defense responses to wounding and Ascochyta rabiei invasion. AB - Wounding chickpea (Cicer arietinum) internodes or cotyledons resulted in an increase in the steady-state level of copper amine oxidase (CuAO) expression both locally and systemically. Dissection of the molecular mechanisms controlling CuAO expression indicated that jasmonic acid worked as a potent inducer of the basal and wound-inducible CuAO expression, whereas salicylic acid and abscisic acid caused a strong reduction of the wound-induced CuAO expression, without having any effect on the basal levels. Epicotyl treatment with the CuAO mechanism-based inhibitor 2-bromoethylamine decreased hydrogen peroxide (H(2)O(2)) levels in all the internodes, as evidenced in vivo by 3,3'-diaminobenzidine oxidation. Moreover, inhibitor pretreatment of wounded epicotyls resulted in a lower accumulation of H(2)O(2) both at the wound site and in distal organs. In vivo CuAO inhibition by 2-bromoethylamine after inoculation of resistant chickpea cv Sultano with Ascochyta rabiei resulted in the development of extended necrotic lesions, with extensive cell damage occurring in sclerenchyma and cortical parenchyma tissues. These results, besides stressing the fine-tuning by key signaling molecules in wound-induced CuAO regulation, demonstrate that local and systemic CuAO induction is essential for H(2)O(2) production in response to wounding and indicate the relevance of these enzymes in protection against pathogens. PMID- 11891245 TI - Digalactosyldiacylglycerol synthesis in chloroplasts of the Arabidopsis dgd1 mutant. AB - Galactolipid biosynthesis in plants is highly complex. It involves multiple pathways giving rise to different molecular species. To assess the contribution of different routes of galactolipid synthesis and the role of molecular species for growth and photosynthesis, we initiated a genetic approach of analyzing double mutants of the digalactosyldiacylglycerol (DGDG) synthase mutant dgd1 with the acyltransferase mutant, act1, and the two desaturase mutants, fad2 and fad3. The double mutants showed different degrees of growth retardation: act1,dgd1 was most severely affected and growth of fad2,dgd1 was slightly reduced, whereas fad3,dgd1 plants were very similar to dgd1. In act1,dgd1, lipid and chlorophyll content were reduced and photosynthetic capacity was affected. Molecular analysis of galactolipid content, fatty acid composition, and positional distribution suggested that the growth deficiency is not caused by changes in galactolipid composition per se. Chloroplasts of dgd1 were capable of synthesizing monogalactosyldiacylglycerol, DGDG, and tri- and tetragalactosyldiacylglycerol. Therefore, the reduced growth of act1,dgd1 and fad2,dgd1 cannot be explained by the absence of DGDG synthase activity from chloroplasts. Molecular analysis of DGDG accumulating in the mutants during phosphate deprivation suggested that similarly to the residual DGDG of dgd1, this additional lipid is synthesized in association with chloroplast membranes through a pathway independent of the mutations, act1, dgd1, fad2, and fad3. Our data imply that the severe growth defect of act1,dgd1 is caused by a reduced metabolic flux of chloroplast lipid synthesis through the eukaryotic and prokaryotic pathway as well as by the reduction of photosynthetic capacity caused by the destabilization of photosynthetic complexes. PMID- 11891246 TI - Comparison of RNA expression profiles based on maize expressed sequence tag frequency analysis and micro-array hybridization. AB - Assembly of 73,000 expressed sequence tags (ESTs) representing multiple organs and developmental stages of maize (Zea mays) identified approximately 22,000 tentative unique genes (TUGs) at the criterion of 95% identity. Based on sequence similarity, overlap between any two of nine libraries with more than 3,000 ESTs ranged from 4% to 20% of the constituent TUGs. The most abundant ESTs were recovered from only one or a minority of the libraries, and only 26 EST contigs had members from all nine EST sets (presumably representing ubiquitously expressed genes). For several examples, ESTs for different members of gene families were detected in distinct organs. To study this further, two types of micro-array slides were fabricated, one containing 5,534 ESTs from 10- to 14-d old endosperm, and the other 4,844 ESTs from immature ear, estimated to represent about 2,800 and 2,500 unique genes, respectively. Each array type was hybridized with fluorescent cDNA targets prepared from endosperm and immature ear poly(A(+)) RNA. Although the 10- to 14-d-old postpollination endosperm TUGs showed only 12% overlap with immature ear TUGs, endosperm target hybridized with 94% of the ear TUGs, and ear target hybridized with 57% of the endosperm TUGs. Incomplete EST sampling of low-abundance transcripts contributes to an underestimate of shared gene expression profiles. Reassembly of ESTs at the criterion of 90% identity suggests how cross hybridization among gene family members can overestimate the overlap in genes expressed in micro-array hybridization experiments. PMID- 11891247 TI - The expression of an extensin-like protein correlates with cellular tip growth in tomato. AB - Extensins are abundant proteins presumed to determine physical characteristics of the plant cell wall. We have cloned a cDNA encoding LeExt1 from a tomato (Lycopersicon esculentum Mill.) root hair cDNA library. The deduced sequence of the LeExt1 polypeptide defined a novel type of extensin-like proteins in tomato. Patterns of mRNA distribution indicated that expression of the LeExt1 gene was initiated in the root hair differentiation zone of the tomato rhizodermis. Cloning of the corresponding promoter and fusion to the -glucuronidase (GUS) reporter gene allowed detailed examination of LeExt1 expression in transgenic tomato plants. Evidence is presented for a direct correlation between LeExt1 expression and cellular tip growth. LeExt1/GUS expression was detectable in trichoblasts (=root hair-bearing cells), but not in atrichoblasts of the tomato rhizodermis. Both hair formation and LeExt1 expression was inducible by the plant hormone ethylene. Comparative analysis of the LeExt1/GUS expression was performed in transgenic tomato, potato (Solanum tuberosum), tobacco (Nicotiana tabacum), and Arabidopsis plants. In the apical/basal dimension, GUS staining was absent from the root cap and undifferentiated cells at the root tip in all species investigated. It was induced at the distal end of the differentiation zone and remained high proximally to the root/hypocotyl boundary. In the radial dimension, GUS expression was root hair specific in the solanaceous species. Whereas LeExt1 mRNA was exclusively detectable in the rhizodermis, root hair-specific expression correlated with GUS expression in germinating pollen tubes. This is correlative evidence for a role of LeExt1 in root hair tip growth [corrected]. PMID- 11891248 TI - The Endoplasmic reticulum-associated maize GL8 protein is a component of the acyl coenzyme A elongase involved in the production of cuticular waxes. AB - The gl8 gene is required for the normal accumulation of cuticular waxes on maize (Zea mays) seedling leaves. The predicted GL8 protein exhibits significant sequence similarity to a class of enzymes that catalyze the reduction of a ketone group to a hydroxyl group. Polyclonal antibodies raised against the recombinant Escherichia coli-expressed GL8 protein were used to investigate the function of this protein in planta. Subcellular fractionation experiments indicate that the GL8 protein is associated with the endoplasmic reticulum membranes. Furthermore, polyclonal antibodies raised against the partially purified leek (Allium porrum) microsomal acyl-coenzyme A (CoA) elongase can react with the E. coli-expressed GL8 protein. In addition, anti-GL8 immunoglobulin G inhibited the in vitro elongation of stearoyl-CoA by leek and maize microsomal acyl-CoA elongase. In combination, these findings indicate that the GL8 protein is a component of the acyl-CoA elongase. In addition, the finding that anti-GL8 immunoglobulin G did not significantly inhibit the 3-ketoacyl-CoA synthase, 3-ketoacyl-CoA dehydrase, and (E) 2,3-enoyl-CoA reductase partial reactions of leek or maize acyl-CoA elongase lends further support to our previous hypothesis that the GL8 protein functions as a beta-ketoacyl reductase during the elongation of very long-chain fatty acids required for the production of cuticular waxes. PMID- 11891249 TI - Identification, purification, and molecular cloning of N-1-naphthylphthalmic acid binding plasma membrane-associated aminopeptidases from Arabidopsis. AB - Polar transport of the plant hormone auxin is regulated at the cellular level by inhibition of efflux from a plasma membrane (PM) carrier. Binding of the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA) to a regulatory site associated with the carrier has been characterized, but the NPA-binding protein(s) have not been identified. Experimental disparities between levels of high-affinity NPA binding and auxin transport inhibition can be explained by the presence of a low-affinity binding site and in vivo hydrolysis of NPA. In Arabidopsis, colocalization of NPA amidase and aminopeptidase (AP) activities, inhibition of auxin transport by artificial beta-naphthylamide substrates, and saturable displacement of NPA by the AP inhibitor bestatin suggest that PM APs may be involved in both low-affinity NPA binding and hydrolysis. We report the purification and molecular cloning of NPA-binding PM APs and associated proteins from Arabidopsis. This is the first report of PM APs in plants. PM proteins were purified by gel permeation, anion exchange, and NPA affinity chromatography monitored for tyrosine-AP activity. Lower affinity fractions contained two orthologs of mammalian APs involved in signal transduction and cell surface extracellular matrix interactions. AtAPM1 and ATAPP1 have substrate specificities and inhibitor sensitivities similar to their mammalian orthologs, and have temporal and spatial expression patterns consistent with previous in planta histochemical data. Copurifying proteins suggest that the APs interact with secreted cell surface and cell wall proline-rich proteins. AtAPM1 and AtAPP1 are encoded by single genes. In vitro translation products of ATAPM1 and AtAPP1 have enzymatic activities similar to those of native proteins. PMID- 11891250 TI - Snakin-2, an antimicrobial peptide from potato whose gene is locally induced by wounding and responds to pathogen infection. AB - The peptide snakin-2 (StSN2) has been isolated from potato (Solanum tuberosum cv Jaerla) tubers and found to be active (EC(50) = 1-20 microM) against fungal and bacterial plant pathogens. It causes a rapid aggregation of both Gram-positive and Gram-negative bacteria. The corresponding StSN2 cDNA encodes a signal sequence followed by a 15-residue acidic sequence that precedes the mature StSN2 peptide, which is basic (isoelectric point = 9.16) and 66 amino acid residues long (molecular weight of 7,025). The StSN2 gene is developmentally expressed in tubers, stems, flowers, shoot apex, and leaves, but not in roots, or stolons, and is locally up-regulated by wounding and by abscisic acid treatment. Expression of this gene is also up-regulated after infection of potato tubers with the compatible fungus Botritys cinerea and down-regulated by the virulent bacteria Ralstonia solanacearum and Erwinia chrysanthemi. These observations are congruent with the hypothesis that the StSN2 is a component of both constitutive and inducible defense barriers. PMID- 11891251 TI - Ethylene enhances water transport in hypoxic aspen. AB - Water transport was examined in solution culture grown seedlings of aspen (Populus tremuloides) after short-term exposures of roots to exogenous ethylene. Ethylene significantly increased stomatal conductance, root hydraulic conductivity (L(p)), and root oxygen uptake in hypoxic seedlings. Aerated roots that were exposed to ethylene also showed enhanced L(p). An ethylene action inhibitor, silver thiosulphate, significantly reversed the enhancement of L(p) by ethylene. A short-term exposure of excised roots to ethylene significantly enhanced the root water flow (Q(v)), measured by pressurizing the roots at 0.3 MPa. The Q(v) values in ethylene-treated roots declined significantly when 50 microM HgCl(2) was added to the root medium and this decline was reversed by the addition of 20 mM 2-mercaptoethanol. The results suggest that the response of Q(v) to ethylene involves mercury-sensitive water channels and that root-absorbed ethylene enhanced water permeation through roots, resulting in an increase in root water transport and stomatal opening in hypoxic seedlings. PMID- 11891252 TI - Ascorbate deficiency can limit violaxanthin de-epoxidase activity in vivo. AB - As a response to high light, plants have evolved non-photochemical quenching (NPQ), mechanisms that lead to the dissipation of excess absorbed light energy as heat, thereby minimizing the formation of dangerous oxygen radicals. One component of NPQ is pH dependent and involves the formation of zeaxanthin from violaxanthin. The enzyme responsible for the conversion of violaxanthin to zeaxanthin is violaxanthin de-epoxidase, which is located in the thylakoid lumen, is activated by low pH, and has been shown to use ascorbate (vitamin C) as its reductant in vitro. To investigate the effect of low ascorbate levels on NPQ in vivo, we measured the induction of NPQ in a vitamin C-deficient mutant of Arabidopsis, vtc2-2. During exposure to high light (1,500 micromol photons m(-2) s(-1)), vtc2-2 plants initially grown in low light (150 micromol photons m(-2) s( 1)) showed lower NPQ than the wild type, but the same quantum efficiency of photosystem II. Crosses between vtc2-2 and Arabidopsis ecotype Columbia established that the ascorbate deficiency cosegregated with the NPQ phenotype. The conversion of violaxanthin to zeaxanthin induced by high light was slower in vtc2-2, and this conversion showed saturation below the wild-type level. Both the NPQ and the pigment phenotype of the mutant could be rescued by feeding ascorbate to leaves, establishing a direct link between ascorbate, zeaxanthin, and NPQ. These experiments suggest that ascorbate availability can limit violaxanthin de epoxidase activity in vivo, leading to a lower NPQ. The results also demonstrate the interconnectedness of NPQ and antioxidants, both important protection mechanisms in plants. PMID- 11891253 TI - Delayed abscission and shorter Internodes correlate with a reduction in the ethylene receptor LeETR1 transcript in transgenic tomato. AB - Stable transformation of tomato (Lycopersicon esculentum cv Ailsa Craig) plants with a construct containing the antisense sequence for the receiver domain and 3' untranslated portion of the tomato ethylene receptor (LeETR1) under the control of an enhanced cauliflower mosaic virus 35S promoter resulted in some expected and unexpected phenotypes. In addition to reduced LeETR1 transcript levels, the two most consistently observed phenotypes in the transgenic lines were delayed abscission and reduced plant size. Fruit coloration and softening were essentially unaffected, and all the seedlings from first generation seed displayed a normal triple response to ethylene. Two independent lines with a single copy of the transgene and reduced LeETR1 transcript accumulation were selected for detailed phenotypic analysis of second generation (R1) plants. Delayed abscission, shorter internode length, and reduced auxin movement all correlated with the presence of the transgene and the degree of reduced LeETR1 transcript accumulation. No significant differences were noted for fruit coloration or fruit softening on R1 plants and all seedlings from R1 and R2 seed displayed a normal triple response. LeETR2 transcript accumulation was only slightly reduced in the R1 plants compared with azygous plants, and LeETR3 (NR) transcript levels appeared to be unaffected by the transgene. We propose that ethylene signal transduction occurs through parallel paths that partially intersect to regulate shared ethylene responses. PMID- 11891254 TI - Isolation of lysophosphatidic acid phosphatase from developing peanut cotyledons. AB - The soluble fraction of immature peanut (Arachis hypogaea) was capable of dephosphorylating [(3)H]lysophosphatidic acid (LPA) to generate monoacylglycerol (MAG). The enzyme responsible for the generation of MAG, LPA phosphatase, has been identified in plants and purified by successive chromatography separations on octyl-Sepharose, Blue Sepharose, Superdex-75, and heparin-agarose to apparent homogeneity from developing peanuts. This enzyme was purified 5,048-fold to a final specific activity of 858 nmol min(-1) mg(-1). The enzyme has a native molecular mass of approximately 39 kD determined by gel filtration and migrates as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a subunit molecular mass of 39 +/- 1.5 kD. The K(m) values for oleoyl-, stearoyl-, and palmitoyl-sn-glycerol-3-phosphate were determined to be 28.6, 39.3, and 47.9 microM, respectively. The LPA phosphatase was specific to LPA and did not utilize any other substrate such as glycerol-3-phosphate, phosphatidic acid, or p-nitrophenylphosphate. The enzyme activity was stimulated by the low concentrations of detergents such as Triton X-100 and octylglucoside. Cations had no effect on the enzyme activity. Fatty acids, sphingosine, and sphingomyelin at low concentrations stimulated the enzyme activity. The identification of LPA phosphatase in plants demonstrates the existence of MAG biosynthetic machinery in plants. PMID- 11891255 TI - Arabidopsis contains ancient classes of differentially expressed actin-related protein genes. AB - Actin-related proteins (ARPs) share less than 60% amino acid sequence homology with conventional actins and have roles in diverse cytoskeletal processes in the cytoplasm and nucleus. The genome of Arabidopsis was explored for possible ARP gene family members. Eight potential ARP gene sequences were found dispersed on three of the five Arabidopsis chromosomes. AtARP2 and AtARP3 are protein orthologs of their similarly named counterparts in other kingdoms. AtARP4, AtARP5, and AtARP6 are orthologs of two classes of nuclear ARPs previously characterized in animals and fungi, BAF53s and ARP6s. AtARP7 and AtARP8 appear to be novel proteins that are not closely related to any known animal or fungal ARPs, and may be plant specific. The complex Arabidopsis ARP gene structures each contain from five to 20 exons. Expressed transcripts were identified and characterized for AtARP2 through AtARP8, but not for AtARP9, and transcripts representing two splice variants were found for AtARP8. The seven expressed genes are predicted to encode proteins ranging from 146 to 471 amino acids in length. Relative to conventional actin and the other ARPs, AtARP2 and AtARP3 transcripts are expressed at very low levels in all organs. AtARP5, AtARP6, and AtARP8 each have distinct transcript expression patterns in seedlings, roots, leaves, flowers, and siliques. Using isovariant-specific monoclonal antibodies, AtARP4 and AtARP7 proteins were shown to be most highly expressed in flowers. The likely involvement of plant ARPs in actin nucleation, branching of actin filaments, chromatin restructuring, and transcription are briefly discussed. PMID- 11891257 TI - Ferrous ion transport across chloroplast inner envelope membranes. AB - The initial rate of Fe(2+) movement across the inner envelope membrane of pea (Pisum sativum) chloroplasts was directly measured by stopped-flow spectrofluorometry using membrane vesicles loaded with the Fe(2+)-sensitive fluorophore, Phen Green SK. The rate of Fe(2+) transport was rapid, coming to equilibrium within 3s. The maximal rate and concentration dependence of Fe(2+) transport in predominantly right-side-out vesicles were nearly equivalent to those measured in largely inside-out vesicles. Fe(2+) transport was stimulated by an inwardly directed electrochemical proton gradient across right-side-out vesicles, an effect that was diminished by the addition of valinomycin in the presence of K(+). Fe(2+) transport was inhibited by Zn(2+), in a competitive manner, as well as by Cu(2+) and Mn(2+). These results indicate that inward directed Fe(2+) transport across the chloroplast inner envelope occurs by a potential-stimulated uniport mechanism. PMID- 11891256 TI - An Arabidopsis calcium-dependent protein kinase is associated with the endoplasmic reticulum. AB - Arabidopsis contains 34 genes that are predicted to encode calcium-dependent protein kinases (CDPKs). CDPK enzymatic activity previously has been detected in many locations in plant cells, including the cytosol, the cytoskeleton, and the membrane fraction. However, little is known about the subcellular locations of individual CDPKs or the mechanisms involved in targeting them to those locations. We investigated the subcellular location of one Arabidopsis CDPK, AtCPK2, in detail. Membrane-associated AtCPK2 did not partition with the plasma membrane in a two-phase system. Sucrose gradient fractionation of microsomes demonstrated that AtCPK2 was associated with the endoplasmic reticulum (ER). AtCPK2 does not contain transmembrane domains or known ER-targeting signals, but does have predicted amino-terminal acylation sites. AtCPK2 was myristoylated in a cell-free extract and myristoylation was prevented by converting the glycine at the proposed site of myristate attachment to alanine (G2A). In plants, the G2A mutation decreased AtCPK2 membrane association by approximately 50%. A recombinant protein, consisting of the first 10 amino acids of AtCPK2 fused to the amino-terminus of beta-glucuronidase, was also targeted to the ER, indicating that the amino terminus of AtCPK2 can specify ER localization of a soluble protein. These results indicate that AtCPK2 is localized to the ER, that myristoylation is likely to be involved in the membrane association of AtCPK2, and that the amino terminal region of AtCPK2 is sufficient for correct membrane targeting. PMID- 11891258 TI - Simultaneous visualization of peroxisomes and cytoskeletal elements reveals actin and not microtubule-based peroxisome motility in plants. AB - Peroxisomes were visualized in living plant cells using a yellow fluorescent protein tagged with a peroxisomal targeting signal consisting of the SKL motif. Simultaneous visualization of peroxisomes and microfilaments/microtubules was accomplished in onion (Allium cepa) epidermal cells transiently expressing the yellow fluorescent protein-peroxi construct, a green fluorescent protein-mTalin construct that labels filamentous-actin filaments, and a green fluorescent protein-microtubule-binding domain construct that labels microtubules. The covisualization of peroxisomes and cytoskeletal elements revealed that, contrary to the reports from animal cells, peroxisomes in plants appear to associate with actin filaments and not microtubules. That peroxisome movement is actin based was shown by pharmacological studies. For this analysis we used onion epidermal cells and various cell types of Arabidopsis including trichomes, root hairs, and root cortex cells exhibiting different modes of growth. In transient onion epidermis assay and in transgenic Arabidopsis plants, an interference with the actin cytoskeleton resulted in progressive loss of saltatory movement followed by the aggregation and a complete cessation of peroxisome motility within 30 min of drug application. Microtubule depolymerization or stabilization had no effect. PMID- 11891259 TI - Benzothiadiazole-induced priming for potentiated responses to pathogen infection, wounding, and infiltration of water into leaves requires the NPR1/NIM1 gene in Arabidopsis. AB - Systemic acquired resistance (SAR) is a plant defense state that is induced, for example, after previous pathogen infection or by chemicals that mimic natural signaling compounds. SAR is associated with the ability to induce cellular defense responses more rapidly and to a greater degree than in noninduced plants, a process called "priming." Arabidopsis plants were treated with the synthetic SAR inducer benzothiadiazole (BTH) before stimulating two prominent cellular defense responses, namely Phe AMMONIA-LYASE (PAL) gene activation and callose deposition. Although BTH itself was essentially inactive at the immediate induction of these two responses, the pretreatment with BTH greatly augmented the subsequent PAL gene expression induced by Pseudomonas syringae pv. tomato infection, wounding, or infiltrating the leaves with water. The BTH pretreatment also enhanced the production of callose, which was induced by wounding or infiltrating the leaves with water. It is interesting that the potentiation by BTH pretreatment of PAL gene activation and callose deposition was not seen in the Arabidopsis nonexpresser of PR genes 1/noninducible immunity 1 mutant, which is compromised in SAR. In a converse manner, augmented PAL gene activation and enhanced callose biosynthesis were found, without BTH pretreatment, in the Arabidopsis constitutive expresser of pathogenesis-related genes (cpr)1 and constitutive expresser of pathogenesis-related genes 5 mutants, in which SAR is constitutive. Moreover, priming for potentiated defense gene activation was also found in pathogen-induced SAR. In sum, the results suggest that priming is an important cellular mechanism in acquired disease resistance of plants that requires the nonexpresser of PR genes 1/noninducible immunity 1 gene. PMID- 11891260 TI - The Arabidopsis phospholipase D family. Characterization of a calcium-independent and phosphatidylcholine-selective PLD zeta 1 with distinct regulatory domains. AB - Four types of phospholipase D (PLD), PLD alpha, beta, gamma, and delta, have been characterized in Arabidopsis, and they display different requirements for Ca(2+), phosphatidylinositol 4,5-bisphosphate (PIP(2)), substrate vesicle composition, and/or free fatty acids. However, all previously cloned plant PLDs contain a Ca(2+)-dependent phospholipid-binding C2 domain and require Ca(2+) for activity. This study documents a new type of PLD, PLD zeta 1, which is distinctively different from previously characterized PLDs. It contains at the N terminus a Phox homology domain and a pleckstrin homology domain, but not the C2 domain. A full-length cDNA for Arabidopsis PLD zeta 1 has been identified and used to express catalytically active PLD in Escherichia coli. PLD zeta 1 does not require Ca(2+) or any other divalent cation for activity. In addition, it selectively hydrolyzes phosphatidylcholine, whereas the other Arabidopsis PLDs use several phospholipids as substrates. PLD zeta 1 requires PIP(2) for activity, but unlike the PIP(2)-requiring PLD beta or gamma, phosphatidylethanolamine is not needed in substrate vesicles. These differences are described, together with a genomic analysis of 12 putative Arabidopsis PLD genes that are grouped into alpha, beta, delta, gamma, and zeta based on their gene architectures, sequence similarities, domain structures, and biochemical properties. PMID- 11891261 TI - The priming of amylose synthesis in Arabidopsis leaves. AB - We investigated the mechanism of amylose synthesis in Arabidopsis leaves using (14)C-labeling techniques. First, we tested the hypothesis that short malto oligosaccharides (MOS) may act as primers for granule-bound starch synthase I. We found increased amylose synthesis in isolated starch granules supplied with ADP[(14)C]glucose (ADP[(14)C]Glc) and MOS compared with granules supplied with ADP[(14)C]Glc but no MOS. Furthermore, using a MOS-accumulating mutant (dpe1), we found that more amylose was synthesized than in the wild type, correlating with the amount of MOS in vivo. When wild-type and mutant plants were tested in conditions where both lines had similar MOS contents, no difference in amylose synthesis was observed. We also tested the hypothesis that branches of amylopectin might serve as the primers for granule-bound starch synthase I. In this model, elongated branches of amylopectin are subsequently cleaved to form amylose. We conducted pulse-chase experiments, supplying a pulse of ADP[(14)C]Glc to isolated starch granules or (14)CO(2) to intact plants, followed by a chase period in unlabeled substrate. We detected no transfer of label from the amylopectin fraction to the amylose fraction of starch either in isolated starch granules or in intact leaves, despite varying the time course of the experiments and using a mutant line (sex4) in which high-amylose starch is synthesized. We therefore find no evidence for amylopectin-primed amylose synthesis in Arabidopsis. We propose that MOS are the primers for amylose synthesis in Arabidopsis leaves. PMID- 11891262 TI - Members of the c1/pl1 regulatory gene family mediate the response of maize aleurone and mesocotyl to different light qualities and cytokinins. AB - We investigated the role of transcription factors (R, SN, C1, and PL) in the regulation of anthocyanin biosynthesis by different light qualities (white, red, blue, and ultraviolet) and by cytokinin in maize (Zea mays). We analyzed anthocyanin accumulation, structural gene expression, and regulatory gene expression in the seed aleurone and the seedling mesocotyl. In the mesocotyl, white, blue, and ultraviolet-B light strongly induced anthocyanin accumulation and expression of two key structural genes. In contrast, red light had little effect. Cytokinin enhanced the response to light but was not sufficient to induce anthocyanin accumulation in darkness. Plants with the pl-bol3 allele showed high levels of anthocyanin accumulation in response to light, whereas those with the pl-W22 allele did not, demonstrating the importance of pl1 in the light response. The expression of the pl-bol3 gene, encoding an MYB-related transcription factor, was induced by light and enhanced by cytokinin in a very similar manner to the structural genes and anthocyanin accumulation. Expression of the bHLH (basic helix-loop-helix) Sn1-bol3 gene was stimulated by several light qualities, but not enhanced by cytokinin, and was less well correlated with the induction of anthocyanin biosynthesis. In the aleurone, white, red, and blue light were effective in stimulating anthocyanin accumulation and expression of the MYB related gene C1. The bHLH R gene was constitutively expressed. We conclude that specific members of the MYB-related c1/pl1 gene family play important roles in the regulation of anthocyanin synthesis in maize in response to different light qualities and cytokinin. PMID- 11891263 TI - Temperature acclimation of photosynthesis and related changes in photosystem II electron transport in winter wheat. AB - Winter wheat (Triticum aestivum L. cv Norin No. 61) was grown at 25 degrees C until the third leaves reached about 10 cm in length and then at 15 degrees C, 25 degrees C, or 35 degrees C until full development of the third leaves (about 1 week at 25 degrees C, but 2-3 weeks at 15 degrees C or 35 degrees C). In the leaves developed at 15 degrees C, 25 degrees C, and 35 degrees C, the optimum temperature for CO(2)-saturated photosynthesis was 15 degrees C to 20 degrees C, 25 degrees C to 30 degrees C, and 35 degrees C, respectively. The photosystem II (PS II) electron transport, determined either polarographically with isolated thylakoids or by measuring the modulated chlorophyll a fluorescence in leaves, also showed the maximum rate near the temperature at which the leaves had developed. Maximum rates of CO(2)-saturated photosynthesis and PS II electron transport determined at respective optimum temperatures were the highest in the leaves developed at 25 degrees C and lowest in the leaves developed at 35 degrees C. So were the levels of chlorophyll, photosystem I and PS II, whereas the level of Rubisco decreased with increasing temperature at which the leaves had developed. Kinetic analyses of chlorophyll a fluorescence changes and P700 reduction showed that the temperature dependence of electron transport at the plastoquinone and water-oxidation sites was modulated by the temperature at which the leaves had developed. These results indicate that the major factor that contributes to thermal acclimation of photosynthesis in winter wheat is the plastic response of PS II electron transport to environmental temperature. PMID- 11891264 TI - The negatively acting factors EID1 and SPA1 have distinct functions in phytochrome A-specific light signaling. AB - EID1 (empfindlicher im dunkelroten Licht) and SPA1 (suppressor of phytochrome A[phyA]-105) function as negatively acting components in phyA-specific light signaling. Mutants in the respective genes led to very similar phenotypes under weak-light conditions. To examine whether both genes are functionally redundant, detailed physiological and genetic analyses were performed with eid1 and spa1 mutants isolated from the same wild-type background. Measurements of hypocotyl elongation, anthocyanin accumulation, and Lhcb1-transcript accumulation under different light treatments demonstrated that SPA1 has a strong influence on the regulation of very low fluence responses and a weaker influence on high irradiance responses. In contrast, EID1 severely altered high-irradiance responses and caused almost no change on very low fluence responses. Analyses on eid1 phyA-105 double mutants demonstrated that EID1 could not suppress the phenotype of the weak phyA allele under continuous far-red light. Measurements on eid1 spa1 double mutants exhibited a strong interference of both genes in the regulation of hypocotyl elongation. These results indicate that EID1 and SPA1 are involved in different but interacting phyA-dependent signal transduction chains. PMID- 11891265 TI - Gamma-glutamyl transpeptidase in transgenic tobacco plants. Cellular localization, processing, and biochemical properties. AB - gamma-Glutamyl transpeptidase (gamma-GT) is a ubiquitous enzyme that catalyzes the first step of glutathione (GSH) degradation in the gamma-glutamyl cycle in mammals. A cDNA encoding an Arabidopsis homolog for gamma-GT was overexpressed in tobacco (Nicotiana tabacum) plants. A high level of the membrane-bound gamma-GT activity was localized outside the cell in transgenic plants. The overproduced enzyme was characterized by a high affinity to GSH and was cleaved post translationally in two unequal subunits. Thus, Arabidopsis gamma-GT is similar to the mammalian enzymes in enzymatic properties, post-translational processing, and cellular localization, suggesting analogous biological functions as a key enzyme in the catabolism of GSH. PMID- 11891266 TI - Uptake kinetics of arsenic species in rice plants. AB - Arsenic (As) finds its way into soils used for rice (Oryza sativa) cultivation through polluted irrigation water, and through historic contamination with As based pesticides. As is known to be present as a number of chemical species in such soils, so we wished to investigate how these species were accumulated by rice. As species found in soil solution from a greenhouse experiment where rice was irrigated with arsenate contaminated water were arsenite, arsenate, dimethylarsinic acid, and monomethylarsonic acid. The short-term uptake kinetics for these four As species were determined in 7-d-old excised rice roots. High affinity uptake (0-0.0532 mM) for arsenite and arsenate with eight rice varieties, covering two growing seasons, rice var. Boro (dry season) and rice var. Aman (wet season), showed that uptake of both arsenite and arsenate by Boro varieties was less than that of Aman varieties. Arsenite uptake was active, and was taken up at approximately the same rate as arsenate. Greater uptake of arsenite, compared with arsenate, was found at higher substrate concentration (low-affinity uptake system). Competitive inhibition of uptake with phosphate showed that arsenite and arsenate were taken up by different uptake systems because arsenate uptake was strongly suppressed in the presence of phosphate, whereas arsenite transport was not affected by phosphate. At a slow rate, there was a hyperbolic uptake of monomethylarsonic acid, and limited uptake of dimethylarsinic acid. PMID- 11891267 TI - Leaf urea metabolism in potato. Urease activity profile and patterns of recovery and distribution of (15)N after foliar urea application in wild-type and urease antisense transgenics. AB - The influence of urease activity on N distribution and losses after foliar urea application was investigated using wild-type and transgenic potato (Solanum tuberosum cv Desiree) plants in which urease activity was down-regulated. A good correlation between urease activity and (15)N urea metabolism (NH(3) accumulation) was found. The general accumulation of ammonium in leaves treated with urea indicated that urease activity is not rate limiting, at least initially, for the assimilation of urea N by the plant. It is surprising that there was no effect of urease activity on either N losses or (15)N distribution in the plants after foliar urea application. Experiments with wild-type plants in the field using foliar-applied (15)N urea demonstrated an initial rapid export of N from urea-treated leaves to the tubers within 48 h, followed by a more gradual redistribution during the subsequent days. Only 10% to 18% of urea N applied was lost (presumably because of NH(3) volatilization) in contrast to far greater losses reported in several other studies. The pattern of urease activity in the canopy was investigated during plant development. The activity per unit protein increased up to 10-fold with leaf and plant age, suggesting a correlation with increased N recycling in senescing tissues. Whereas several reports have claimed that plant urease is inducible by urea, no evidence for urease induction could be found in potato. PMID- 11891268 TI - Limits to sulfur accumulation in transgenic lupin seeds expressing a foreign sulfur-rich protein. AB - The low sulfur amino acid content of legume seeds restricts their nutritive value for animals. We have investigated the limitations to the accumulation of sulfur amino acids in the storage proteins of narrow leaf lupin (Lupinus angustifolius) seeds. Variation in sulfur supply to lupin plants affected the sulfur amino acid accumulation in the mature seed. However, when sulfur was in abundant supply, it accumulated to a large extent in oxidized form, rather than reduced form, in the seeds. At all but severely limiting sulfur supply, addition of a transgenic (Tg) sink for organic sulfur resulted in an increase in seed sulfur amino acid content. We hypothesize that demand, or sink strength for organic sulfur, which is itself responsive to environmental sulfur supply, was the first limit to the methionine (Met) and cysteine (Cys) content of wild-type lupin seed protein under most growing conditions. In Tg, soil-grown seeds expressing a foreign Met- and Cys-rich protein, decreased pools of free Met, free Cys, and glutathione indicated that the rate of synthesis of sulfur amino acids in the cotyledon had become limiting. Homeostatic mechanisms similar to those mediating the responses of plants to environmental sulfur stress resulted in an adjustment of endogenous protein composition in Tg seeds, even when grown at adequate sulfur supply. Uptake of sulfur by lupin cotyledons, as indicated by total seed sulfur at maturity, responded positively to increased sulfur supply, but not to increased demand in the Tg seeds. PMID- 11891270 TI - Complex pattern of Mycobacterium marinum gene expression during long-term granulomatous infection. AB - During latent infection of humans with Mycobacterium tuberculosis, bacteria persist in the asymptomatic host within granulomas, organized collections of differentiated macrophages, and other immune cells. The mechanisms for persistence remain poorly understood, as is the metabolic and replicative state of the microbes within granulomas. We analyzed the gene expression profile of Mycobacterium marinum, the cause of fish and amphibian tuberculosis, during its persistence in granulomas. We identified genes expressed specifically when M. marinum persists within granulomas. These granuloma-activated genes were not activated in vitro in response to various conditions postulated to be operant in tuberculous granulomas, suggesting that their granuloma-specific activation was caused by complex conditions that could not be mimicked in vitro. In addition to the granuloma-activated genes, the bacteria resident in granulomas expressed a wide range of metabolic and synthetic genes that are expressed during logarithmic growth in laboratory medium. Our results suggest a dynamic host-pathogen interaction in the granuloma, where metabolically active bacteria are kept in check by the host immune system and where the products of granuloma-specific bacterial genes may thwart the host's attempt to completely eradicate the bacteria. PMID- 11891269 TI - Changes in the expression and the enzymic properties of the 20S proteasome in sugar-starved maize roots. evidence for an in vivo oxidation of the proteasome. AB - The 20S proteasome (multicatalytic proteinase) was purified from maize (Zea mays L. cv DEA 1992) roots through a five-step procedure. After biochemical characterization, it was shown to be similar to most eukaryotic proteasomes. We investigated the involvement of the 20S proteasome in the response to carbon starvation in excised maize root tips. Using polyclonal antibodies, we showed that the amount of proteasome increased in 24-h-carbon-starved root tips compared with freshly excised tips, whereas the mRNA levels of alpha 3 and beta 6 subunits of 20S proteasome decreased. Moreover, in carbon-starved tissues, chymotrypsin like and caseinolytic activities of the 20S proteasome were found to increase, whereas trypsin-like activities decreased. The measurement of specific activities and kinetic parameters of 20S proteasome purified from 24-h-starved root tips suggested that it was subjected to posttranslational modifications. Using dinitrophenylhydrazine, a carbonyl-specific reagent, we observed an increase in carbonyl residues in 20S proteasome purified from starved root tips. This means that 20S proteasome was oxidized during starvation treatment. Moreover, an in vitro mild oxidative treatment of 20S proteasome from non-starved material resulted in the activation of chymotrypsin-like, peptidyl-glutamyl-peptide hydrolase and caseinolytic-specific activities and in the inhibition of trypsin like specific activities, similar to that observed for proteasome from starved root tips. Our results provide the first evidence, to our knowledge, for an in vivo carbonylation of the 20S proteasome. They suggest that sugar deprivation induces an oxidative stress, and that oxidized 20S proteasome could be associated to the degradation of oxidatively damaged proteins in carbon starvation situations. PMID- 11891271 TI - Identification of gene function and functional pathways by systemic plasmid-based ribozyme targeting in adult mice. AB - To date, functional genomic studies have been confined to either cell-based assays or germline mutations, using transgenic or knockout animals. However, these approaches are often unable either to recapitulate complex biologic phenotypes, such as tumor metastasis, or to identify the specific genes and functional pathways that produce serious diseases in adult animals. Although the transcription factor NF-kappaB transactivates many metastasis-related genes in cells, the precise genes and functional-pathways through which NF-kappaB regulates metastasis in tumor-bearing hosts are poorly understood. Here, we show that the systemic delivery of plasmid-based ribozymes targeting NF-kappaB in adult, tumor-bearing mice suppressed NF-kappaB expression in metastatic melanoma cells, as well as in normal cell types, and significantly reduced metastatic spread. Plasmid-based ribozymes suppressed target-gene expression with sequence specificity not achievable by using synthetic oligonucleotide-based approaches. NF-kappaB seemed to regulate tumor metastasis through invasion-related, rather than angiogenesis-, cell-cycle- or apoptosis-related pathways in tumor-bearing mice. Furthermore, ribozymes targeting either of the NF-kappaB-regulated genes, integrin beta(3) or PECAM-1 (a ligand-receptor pair linked to cell adhesion), reduced tumor metastasis at a level comparable to NF-kappaB. These studies demonstrate the utility of gene targeting by means of systemic, plasmid-based ribozymes to dissect out the functional genomics of complex biologic phenotypes, including tumor metastasis. PMID- 11891273 TI - The use of dimetal building blocks in convergent syntheses of large arrays. AB - Within the broad field called "supramolecular chemistry," there is a sector that is based on the use of metal atoms or ions as key elements in promoting the assembly and dictating the main structural features of the supramolecular products. Considerable success has been achieved by using MM bonded dimetal entities in this role. Metal-metal bonded cationic complexes of the [M(2)(DAniF)(n)(MeCN)(8-2n)]((4-n)+) type, where M = Mo or Rh and DAniF is an N,N'-di-p-anisylformamidinate anion, have been used as subunit precursors and then linked by various equatorial and axial bridging groups such as polycarboxylate anions, polypyridyls, and polynitriles. Characterization of the products by single-crystal x-ray diffraction, cyclic voltammetry, differential pulse voltammetry, NMR, and other spectroscopic techniques has revealed the presence of discrete tetranuclear (pairs or loops), hexanuclear (triangles), octanuclear (squares), and dodecanuclear (cages) species and one-, two-, or three dimensional molecular nanotubes. These compounds display a rich electrochemical behavior that is affected by the nature of the linkers. PMID- 11891272 TI - Disruption of estrogen receptor beta gene impairs spatial learning in female mice. AB - Here we provide the first evidence, to our knowledge, that estradiol (E(2)) affects learning and memory via the newly discovered estrogen receptor beta (ERbeta). In this study, ERbeta knockout (ERbetaKO) and wild-type littermates were tested for spatial learning in the Morris water maze after ovariectomy, appropriate control treatment, or one of two physiological doses of E(2). Regardless of treatment, all wild-type females displayed significant learning. However, ERbetaKOs given the low dose of E(2) were delayed in learning acquisition, and ERbetaKOs administered the higher dose of E(2) failed to learn the task. These data show that ERbeta is required for optimal spatial learning and may have implications for hormone replacement therapy in women. PMID- 11891274 TI - Targeting of both mouse neuropilin-1 and neuropilin-2 genes severely impairs developmental yolk sac and embryonic angiogenesis. AB - Neuropilins (NP1 and NP2) are vascular endothelial growth factor (VEGF) receptors that mediate developmental and tumor angiogenesis. Transgenic mice, in which both NP1 and NP2 were targeted (NP1(-/-)NP2(-/-)) died in utero at E8.5. Their yolk sacs were totally avascular. Mice deficient for NP2 but heterozygous for NP1 (NP1(+/-)NP2(-/-)) or deficient for NP1 but heterozygous for NP2 (NP1(-/-)NP2(+/ )) were also embryonic lethal and survived to E10-E10.5. The E10 yolk sacs and embryos were easier to analyze for vascular phenotype than the fragile poorly formed 8.5 embryos. The vascular phenotypes of these E10 mice were very abnormal. The yolk sacs, although of normal size, lacked the larger collecting vessels and had less dense capillary networks. PECAM staining of yolk sac endothelial cells showed the absence of branching arteries and veins, the absence of a capillary bed, and the presence of large avascular spaces between the blood vessels. The embryos displayed blood vessels heterogeneous in size, large avascular regions in the head and trunk, and blood vessel sprouts that were unconnected. The embryos were about 50% the length of wild-type mice and had multiple hemorrhages. These double NP1/NP2 knockout mice had a more severe abnormal vascular phenotype than either NP1 or NP2 single knockouts. Their abnormal vascular phenotype resembled those of VEGF and VEGFR-2 knockouts. These results suggest that NRPs are early genes in embryonic vessel development and that both NP1 and NP2 are required. PMID- 11891275 TI - An unusual Fc receptor-related protein expressed in human centroblasts. AB - Here, we report the identification of Fc receptor homolog expressed in B cells (FREB), a unique B cell-specific molecule that is distantly related to FcgammaRI (receptor I for the Fc fragment of IgG) and is encoded on human chromosome 1q, within the FcgammaR gene region. FREB has an intracellular distribution and lacks a canonical transmembrane domain. In addition, FREB lacks bona fide Fc fragment binding regions and does not bind immunoglobulins. By using specific monoclonal antibodies, we show that FREB is preferentially expressed in germinal center centroblasts, which undergo affinity maturation and class-switch recombination. Together, these characteristics indicate that FREB may have a unique role in B cell differentiation. FREB is also expressed in some B cell lymphomas, most of which have centroblast origin. Remarkably, FREB is expressed in a subset of diffuse large B cell lymphomas, providing a unique marker for the characterization of this B cell malignancy. PMID- 11891276 TI - Vanadate trapping of nucleotide at the ATP-binding sites of human multidrug resistance P-glycoprotein exposes different residues to the drug-binding site. AB - The human multidrug resistance P-glycoprotein uses ATP to transport a wide variety of structurally unrelated cytotoxic compounds out of the cell. In this study, we used cysteine-scanning mutagenesis and cross-linking studies to identify residues that are exposed to the drug-binding site upon vanadate trapping. In the absence of nucleotides, C222(TM4) was cross-linked to C868(TM10) and C872(TM10); C306(TM5) was cross-linked to C868(TM10), C872(TM10), C945(TM11), C982(TM12), and C984(TM12); and C339(TM6) was cross-linked to C868(TM10), C872(TM10), C942(TM11), C982(TM12), and C985(TM12). These cysteines are in the middle of the predicted transmembrane (TM) segments and form the drug-binding site. Cross-linking between 332C(TM6) and cysteines introduced at the extracellular side of other TM segments was also done. In the absence of nucleotides, residues 332C and 856C on the extracellular side of TMs 6 and 10, respectively, were cross-linked with a 13-A cross-linker (M8M, 3,6-dioxaoctane 1,8-diyl bismethanethiosulfonate). ATP plus vanadate inhibited cross-linking between 332C(TM6) and 856C(TM10) as well as those in the drug-binding site. Instead, vanadate trapping promoted cross-linking between 332C(TM6) and 976C(TM12) with a 10-A cross-linker (M6M, 1,6-hexanediyl bismethanethiosulfonate). When ATP hydrolysis was allowed to proceed, then 332C(TM12) could form a disulfide bond with 975C(TM12). The cross-linking pattern of 332C(TM6) with residues in TM10 and TM12 indicates that the drug-binding site undergoes dynamic and relatively large conformational changes, and that different residues are exposed to the drug-binding site during the resting phase, upon vanadate trapping and at the completion of the catalytic cycle. PMID- 11891278 TI - Latent sensitivity to Fas-mediated apoptosis after CD40 ligation may explain activity of CD154 gene therapy in chronic lymphocytic leukemia. AB - Patients with chronic lymphocytic leukemia (CLL) treated with adenovirus (Ad) CD154 (CD40L) gene therapy experience reductions in leukemia cell counts and lymph node size associated with induction of the death receptor Fas (CD95). CD4 T cell lines can induce apoptosis of CD40-activated CLL cells via a CD95 ligand (CD95-L)-dependent mechanism. To examine whether CD95-L was sufficient to induce cytolysis of CD40-activated CLL cells, we used Chinese hamster ovary cells transfected with CD95-L as cytotoxic effector cells. CD40-activated CLL cells were initially resistant to CD95-mediated apoptosis despite high-level expression of CD95. However, after 72 h, CLL cells from seven of seven patients became increasingly sensitive to CD95-mediated apoptosis. This sensitivity correlated with a progressive decline in Flice-inhibitory protein (FLIP), which was induced within 24 h of CD40 ligation. Down-regulation of FLIP with an antisense oligonucleotide or a pharmacologic agent, however, was not sufficient to render CLL cells sensitive to CD95-mediated apoptosis in the 24-72 h after CD40 activation. Although the levels of pro-Caspase-8 appeared sufficient, inadequate levels of Fas-associated death domain protein (FADD) and DAP3 may preclude assembly of the death-inducing signaling complex. Seventy-two hours after CD40 ligation, sensitivity to CD95 and a progressive increase in FADD and DAP3 were associated with the acquired ability of FADD and FLIP to coimmunoprecipitate with the death-inducing signaling complex after CD95 ligation. Collectively, these studies reveal that CD40 ligation on CLL B cells induces a programmed series of events in which the cells initially are protected and then sensitized to CD95 mediated apoptosis through shifts in the balance of the anti- and proapoptotic proteins FLIP and FADD. PMID- 11891277 TI - Transmembrane pores formed by synthetic p-octiphenyl beta-barrels with internal carboxylate clusters: regulation of ion transport by pH and Mg(2+)- complexed 8 aminonaphthalene-1,3,6-trisulfonate. AB - Design, synthesis, and study of a synthetic barrel-stave supramolecule with p octiphenyl "staves," beta-sheet "hoops," and hydrophobic exterior as well as internal carboxylate clusters are reported. Ion transport experiments indicate the formation of transmembrane pores at 5 < pH < 7 with nanomolar activity. Blockage of dye efflux from spherical bilayers by external Mg(OAc)(2) and internal 8-aminonaphthalene-1,3,6-trisulfonate is suggestive for weakly cooperative (n = 1.16) formation of aspartate-Mg(2+)-8-aminonaphthalene-1,3,6 trisulfonate complexes within the barrel-stave supramolecule (K(D) = 2.9 mM). Corroborative evidence from structural studies by circular dichroism spectroscopy is provided and discussed with emphasis of the importance of internal charge repulsion for pore formation and future applications toward binding and catalysis within supramolecular synthetic pores. PMID- 11891279 TI - Controlled disassembling of self-assembling systems: toward artificial molecular level devices and machines. AB - Investigations on self-assembling/induced-disassembling systems have led to the design of molecular-level devices capable of performing a variety of functions. Some of the work carried out in this field is illustrated. PMID- 11891280 TI - Element-specific localization of Drosophila retrotransposon Gag proteins occurs in both nucleus and cytoplasm. AB - Many Drosophila non-long terminal repeat (LTR) retrotransposons actively transpose into internal, gene-rich regions of chromosomes but do not transpose onto chromosome ends. HeT-A and TART are remarkable exceptions; they form telomeres of Drosophila by repeated transpositions onto the ends of chromosomes and never transpose to internal regions of chromosomes. Both telomeric and nontelomeric, non-LTR elements transpose by target-primed reverse transcription, and their targets are not determined simply by DNA sequence, so it is not clear why these two kinds of elements have nonoverlapping transposition patterns. To explore roles of retrotransposon-encoded proteins in transposition, we analyzed intracellular targeting of Gag proteins from five non-LTR retrotransposons, HeT A, TART, jockey, Doc, and I factor. All were expressed as green fluorescent protein-tagged proteins in cultured Drosophila cells. These Gag proteins have high levels of sequence similarity, but they have dramatic differences in intracellular targeting. As expected, HeT-A and TART Gags are transported efficiently to nuclei, where they show specific patterns of localization. These patterns are cell cycle-dependent, disappearing during mitosis. In contrast, only a fraction of jockey Gag moves into nuclei, whereas neither Doc nor I factor Gag is detected in the nucleus. Gags of the nontelomeric retrotransposons form characteristic clusters in the cytoplasm. These experiments demonstrate that closely related retrotransposon Gag proteins can have different intracellular localizations, presumably because they interact differently with cellular components. We suggest that these interactions reflect mechanisms by which the cell influences the level of transposition of an element. PMID- 11891281 TI - Entropically driven self-assembly of multichannel rosette nanotubes. AB - Rosette nanotubes are a new class of organic nanotubes obtained through the hierarchical self-assembly of low molecular weight synthetic modules in water. Here we demonstrate that these materials can serve as scaffolds for the supramolecular synthesis of multichannel nanotubular architectures and report on the discovery of their entropy-driven self-assembly process. PMID- 11891282 TI - Covalent modification of proteins by cocaine. AB - Cocaine covalently modifies proteins through a reaction in which the methyl ester of cocaine acylates the epsilon-amino group of lysine residues. This reaction is highly specific in vitro, because no other amino acid reacts with cocaine, and only cocaine's methyl ester reacts with the lysine side chain. Covalently modified proteins were present in the plasma of rats and human subjects chronically exposed to cocaine. Modified endogenous proteins are immunogenic, and specific antibodies were elicited in mouse and detected in the plasma of human subjects. Covalent modification of proteins could explain cocaine's autoimmune effects and provide a new biochemical approach to cocaine's long-term actions. PMID- 11891283 TI - Genetic variation at the 22q11 PRODH2/DGCR6 locus presents an unusual pattern and increases susceptibility to schizophrenia. AB - The location of a schizophrenia susceptibility locus at chromosome 22q11 has been suggested by genome-wide linkage studies. Additional support was provided by the observation of a higher-than-expected frequency of 22q11 microdeletions in patients with schizophrenia and the demonstration that approximately 20-30% of individuals with 22q11 microdeletions develop schizophrenia or schizoaffective disorder in adolescence and adulthood. Analysis of the extent of these microdeletions by using polymorphic markers afforded further refinement of this locus to a region of approximately 1.5 Mb. Recently, a high rate of 22q11 microdeletions was also reported for a cohort of 47 patients with Childhood Onset Schizophrenia, a rare and severe form of schizophrenia with onset by age 13. It is therefore likely that this 1.5-Mb region contains one or more genes that predispose to schizophrenia. In three independent samples, we provide evidence for a contribution of the PRODH2/DGCR6 locus in 22q11-associated schizophrenia. We also uncover an unusual pattern of PRODH2 gene variation that mimics the sequence of a linked pseudogene. Several of the pseudogene-like variants we identified result in missense changes at conserved residues and may prevent synthesis of a fully functional enzyme. Our results have implications for understanding the genetic basis of the 22q11-associated psychiatric phenotypes and provide further insights into the genomic instability of this region. PMID- 11891284 TI - UBE1L is a retinoid target that triggers PML/RARalpha degradation and apoptosis in acute promyelocytic leukemia. AB - All-trans-retinoic acid (RA) treatment induces remissions in acute promyelocytic leukemia (APL) cases expressing the t(15;17) product, promyelocytic leukemia (PML)/RA receptor alpha (RARalpha). Microarray analyses previously revealed induction of UBE1L (ubiquitin-activating enzyme E1-like) after RA treatment of NB4 APL cells. We report here that this occurs within 3 h in RA-sensitive but not RA-resistant APL cells, implicating UBE1L as a direct retinoid target. A 1.3-kb fragment of the UBE1L promoter was capable of mediating transcriptional response to RA in a retinoid receptor-selective manner. PML/RARalpha, a repressor of RA target genes, abolished this UBE1L promoter activity. A hallmark of retinoid response in APL is the proteasome-dependent PML/RARalpha degradation. UBE1L transfection triggered PML/RARalpha degradation, but transfection of a truncated UBE1L or E1 did not cause this degradation. A tight link was shown between UBE1L induction and PML/RARalpha degradation. Notably, retroviral expression of UBE1L rapidly induced apoptosis in NB4 APL cells, but not in cells lacking PML/RARalpha expression. UBE1L has been implicated directly in retinoid effects in APL and may be targeted for repression by PML/RARalpha. UBE1L is proposed as a direct pharmacological target that overcomes oncogenic effects of PML/RARalpha by triggering its degradation and signaling apoptosis in APL cells. PMID- 11891285 TI - Ion channel gating: a first-passage time analysis of the Kramers type. AB - The opening rate of voltage-gated potassium ion channels exhibits a characteristic knee-like turnover where the common exponential voltage dependence changes suddenly into a linear one. An explanation of this puzzling crossover is put forward in terms of a stochastic first passage time analysis. The theory predicts that the exponential voltage dependence correlates with the exponential distribution of closed residence times. This feature occurs at large negative voltages when the channel is predominantly closed. In contrast, the linear part of voltage dependence emerges together with a nonexponential distribution of closed dwelling times with increasing voltage, yielding a large opening rate. Depending on the parameter set, the closed-time distribution displays a power law behavior that extends over several decades. PMID- 11891286 TI - A robust inducible-repressible promoter greatly facilitates gene knockouts, conditional expression, and overexpression of homologous and heterologous genes in Tetrahymena thermophila. AB - The Cd(2+)-inducible metallothionein (MTT1) gene was cloned from Tetrahymena thermophila. Northern blot analysis showed that MTT1 mRNA is not detectable in the absence of Cd(2+), is induced within 10 min of its addition, is expressed in proportion to its concentration, and rapidly disappears upon its withdrawal. Similarly, when the neo1 gene coding region flanked by the MTT1 gene noncoding sequences was used to disrupt the MTT1 locus, no transformants were observed in the absence of Cd(2+), and the number of transformants was proportional to increased Cd(2+) concentration. The neo3 cassette, in which the MTT1 promoter replaced the histone gene HHF1 promoter of the previously used neo2 cassette, transformed cells at much higher frequencies than neo2 and produced germ-line knockouts where neo2 had failed. Rescuing the progeny of a mating of gamma tubulin gene, GTU1, knockout heterokaryons with a GTU1 gene inserted into the MTT1 locus yielded >75 times more transformants than rescuing with the wild-type GTU1 gene itself. When cells rescued with the MTT1-GTU1 chimeric gene were transferred to medium lacking Cd(2+), they stopped growing and had phenotypic changes indistinguishable from cells containing only disrupted GTU1 genes. Thus, it is now possible to create conditional lethal mutants and study the terminal phenotypes of null mutations for essential genes by replacing the endogenous gene with one under the control of the MTT1 promoter. The MTT1 promoter also resulted in approximately 30 times more overexpression of the IAG48[G1] surface antigen gene of the ciliate fish parasite Ichthyophthirius multifiliis than the highly expressed BTU1 promoter, accounting for approximately 1% of the total cell protein. Thus, the MTT1 promoter should enable routine over-expression of endogenous and foreign genes in Tetrahymena. PMID- 11891287 TI - Ionic dependence of Ca2+ channel modulation by syntaxin 1A. AB - Alteration of the kinetic properties of voltage-gated Ca(2+) channels, Ca(v)1.2 (Lc-type), Ca(v)2.2 (N type), and Ca(v)2.3 (R type), by syntaxin 1A (Syn1A) and synaptotagmin could modulate exocytosis. We tested how switching divalent charge carriers from Ca(2+) to Sr(2+) and Ba(2+) affected Syn1A and synaptotagmin modulation of Ca(2+)-channel activation. Syn1A accelerated Ca(v)1.2 activation if Ca(2+) was the charge carrier; and by substituting for Ba(2+), Syn1A slowed Ca(v)1.2 activation. Syn1A also significantly accelerated Ca(v)2.3 activation in Ca(2+) and marginally in Ba(2+). Synaptotagmin, on the other hand, increased the rate of activation of Ca(v)2.3 and Ca(v)2.2 in all permeating ions tested. The Syn1A-channel interaction, unlike the synaptotagmin-channel interaction, proved significantly more sensitive to the type of permeating ion. It is well established that exocytosis is affected by switching the charge carriers. Based on the present results, we suggest that the channel-Syn1A interaction could respond to the conformational changes induced within the channel during membrane depolarization and divalent ion binding. These changes could partially account for the charge specificity of synaptic transmission as well as for the fast signaling between the Ca(2+) source and the fusion apparatus of channel associated-vesicles (CAV). Furthermore, propagation of conformational changes induced by the divalent ions appear to affect the concerted interaction of the channel with the fusion/docking machinery upstream to free Ca(2+) buildup and/or binding to a cytosolic Ca(2+) sensor. These results raise the intriguing possibility that the channel is the Ca(2+) sensor in the process of fast neurotransmitter release. PMID- 11891288 TI - Notch receptor cleavage depends on but is not directly executed by presenilins. AB - Notch receptors undergo three distinct proteolytic cleavages during maturation and activation. The third cleavage occurs within the plasma membrane and results in the release and translocation of the intracellular domain into the nucleus to execute Notch signaling. This so-called gamma-secretase cleavage is under the control of presenilins, but it is not known whether presenilins themselves carry out the cleavage or whether they act by means of yet-unidentified gamma secretase(s). In this article, we show that Notch intracellular cleavage in intact cells completely depends on presenilins. In contrast, partial purification of the Notch cleavage activity reveals an activity, which is present only in protein extracts from presenilin-containing cells, and which does not comigrate with presenilin. This finding provides evidence for the existence of a specific Notch-processing activity, which is physically distinct from presenilins. We conclude from these experiments that presenilins are critically required for Notch intracellular cleavage but are not themselves directly mediating the cleavage. PMID- 11891289 TI - The role of the dynamics of focal adhesion kinase in the mechanotaxis of endothelial cells. AB - The migration of vascular endothelial cells (ECs) is critical in vascular remodeling. We showed that fluid shear stress enhanced EC migration in flow direction and called this "mechanotaxis." To visualize the molecular dynamics of focal adhesion kinase (FAK) at focal adhesions (FAs), FAK tagged with green fluorescence protein (GFP) was expressed in ECs. Within 10 min of shear stress application, lamellipodial protrusion was induced at cell periphery in the flow direction, with the recruitment of FAK at FAs. ECs under flow migrated with polarized formation of new FAs in flow direction, and these newly formed FAs subsequently disassembled after the rear of the cell moved over them. The cells migrating under flow had a decreased number of FAs. In contrast to shear stress, serum did not significantly affect the speed of cell migration. Serum induced lamellipodia and FAK recruitment at FAs without directional preference. FAK(Y397) phosphorylation colocalized with GFP-FAK at FAs in both shear stress and serum experiments. The total level of FAK(Y397) phosphorylation after shear stress was lower than that after serum treatment, suggesting that the polarized change at cell periphery rather than the total level of FAK(Y397) phosphorylation is important for directional migration. Our results demonstrate the dynamics of FAK at FAs during the directional migration of EC in response to mechanical force, and suggest that mechanotaxis is an important mechanism controlling EC migration. PMID- 11891290 TI - Glial protein S100B modulates long-term neuronal synaptic plasticity. AB - Glial cells are traditionally regarded as elements for structural support and ionic homeostasis, but have recently attracted attention as putative integral elements of the machinery involved in synaptic transmission and plasticity. Here, we demonstrate that calcium-binding protein S100B, which is synthesized in considerable amounts in astrocytes (a major glial cell subtype), modulates long term synaptic plasticity. Mutant mice devoid of S100B developed normally and had no detectable abnormalities in the cytoarchitecture of the brain. These mutant mice, however, had strengthened synaptic plasticity as identified by enhanced long-term potentiation (LTP) in the hippocampal CA1 region. Perfusion of hippocampal slices with recombinant S100B proteins reversed the levels of LTP in the mutant slices to those of the wild-type slices, indicating that S100B might act extracellularly. In addition to enhanced LTP, mutant mice had enhanced spatial memory in the Morris water maze test and enhanced fear memory in the contextual fear conditioning. The results indicate that S100B is a glial modulator of neuronal synaptic plasticity and strengthen the notion that glial neuronal interaction is important for information processing in the brain. PMID- 11891291 TI - Abnormal accumulation of hyaluronan matrix diminishes contact inhibition of cell growth and promotes cell migration. AB - Elevated hyaluronan biosynthesis and matrix deposition correlates with cell proliferation and migration. We ectopically expressed three isoforms of hyaluronan synthase (HAS1, HAS2, or HAS3) in nontransformed rat 3Y1 cells and observed a de novo, massive formation of a hyaluronan matrix that resulted in a partial loss of contact-mediated inhibition of cell growth and migration. All three HAS transfectants showed an enhanced motility in scratch wound assays, and a significant increase in their confluent cell densities. In high-density cultures, the HAS transfectants had a fibroblastic cell shape and markedly formed overlapping cell layers. This phenotype was more pronounced in the HAS2 transfectants than HAS1 or HAS3 transfectants, and occurred with significant alterations in the microfilament organization and N-cadherin distribution at the cell-cell border. Inhibition of a phosphatidylinositol 3-kinase (PI3-kinase) pathway resulted in reacquisition of the normal phenotype of HAS2 transfectants, suggesting that the intracellular PI3-kinase signaling regulates diminution of contact inhibition induced by formation of the massive hyaluronan matrix. Our observations suggest that hyaluronan and its matrix can modulate contact inhibition of cell growth and migration, and provide evidence for functional differences between hyaluronan synthesized by the different HAS proteins. PMID- 11891292 TI - Community-wide distribution of predator-prey interaction strength in kelp forests. AB - The strength of interactions between predators and their prey (interaction strength) varies enormously among species within ecological communities. Understanding the community-wide distribution of interaction strengths is vital, given that communities dominated by weak interactions may be more stable and resistant to invasion. In the oceans, previous studies have reported log-normal distributions of per capita interaction strength. We estimated the distribution of predator-prey interaction strengths within a subtidal speciose herbivore community (45 species). Laboratory experiments were used to determine maximum per capita interaction strengths for eight species of herbivores (including amphipods, isopods, gastropods, and sea urchins) that graze on giant kelp (Macrocystis pyrifera) microscopic stages. We found that maximum per capita interaction strength saturated as a function of individual herbivore biomass, likely caused by predator/prey size thresholds. Incorporating this nonlinearity, we predicted maximum per capita interaction strength for the remaining herbivore species. The resulting distribution of per capita interaction strengths was bimodal, in striking contrast to previous reports from other communities. Although small herbivores often had per capita interaction strengths similar to larger herbivores, their tendency to have greater densities in the field increased their potential impact as grazers. These results indicate that previous conclusions about the distributions of interaction strength in natural communities are not general, and that intermediate-sized predators can under realistic circumstances represent the most effective consumers in natural communities. PMID- 11891293 TI - The tRNA specificity of Thermus thermophilus EF-Tu. AB - By introducing a GAC anticodon, 21 different Escherichia coli tRNAs were misacylated with either phenylalanine or valine and assayed for their affinity to Thermus thermophilus elongation factor Tu (EF-Tu)*GTP by using a ribonuclease protection assay. The presence of a common esterified amino acid permits the thermodynamic contribution of each tRNA body to the overall affinity to be evaluated. The E. coli elongator tRNAs exhibit a wide range of binding affinities that varied from -11.7 kcal/mol for Val-tRNA(Glu) to -8.1 kcal/mol for Val tRNA(Tyr), clearly establishing EF-Tu*GTP as a sequence-specific RNA-binding protein. Because the ionic strength dependence of k(off) varied among tRNAs, some of the affinity differences are the results of a different number of phosphate contacts formed between tRNA and protein. Because EF-Tu is known to contact only the phosphodiester backbone of tRNA, the observed specificity must be a consequence of an indirect readout mechanism. PMID- 11891294 TI - Direct external imaging of nascent cancer, tumor progression, angiogenesis, and metastasis on internal organs in the fluorescent orthotopic model. AB - Mouse tumor models have undergone profound improvements in the fidelity of emulating human disease. Replacing ectopic s.c. implantation with organ-specific orthotopic implantation reproduces human tumor growth and metastasis. Strong fluorescent labeling with green fluorescent protein along with inexpensive video detectors, positioned externally to the mouse, allows the monitoring of details of tumor growth, angiogenesis, and metastatic spread. However, the sensitivity of external imaging is limited by light scattering in intervening tissue, most especially in skin. Opening a reversible skin-flap in the light path markedly reduces signal attenuation, increasing detection sensitivity many-fold. The observable depth of tissue is thereby greatly increased and many tumors that were previously hidden are now clearly observable. This report presents tumor images and related quantitative growth data previously impossible to obtain. Single tumor cells, expressing green fluorescent protein, were seeded on the brain image through a scalp skin-flap. Lung tumor microfoci representing a few cells are viewed through a skin-flap over the chest wall, while contralateral micrometastases were imaged through the corresponding skin-flap. Pancreatic tumors and their angiogenic microvessels were imaged by means of a peritoneal wall skin-flap. A skin-flap over the liver allowed imaging of physiologically relevant micrometastases originating in an orthotopically implanted tumor. Single tumor cells on the liver arising from intraportal injection also were detectable. Possible future technical developments are suggested by the image, through a lower-abdominal skin-flap, of an invasive prostate tumor expressing both red and green fluorescent proteins in separate colonies. PMID- 11891295 TI - A Gaussian-chain model for treating residual charge-charge interactions in the unfolded state of proteins. AB - Characterization of the unfolded state is essential for understanding the protein folding problem. In the unfolded state, a protein molecule samples vastly different conformations. Here I present a simple theoretical method for treating residual charge-charge interactions in the unfolded state. The method is based on modeling an unfolded protein as a Gaussian chain. After sampling over all conformations, the electrostatic interaction energy between two charged residues (separated by l peptide bonds) is given by W = 332(6/pi)(1/2)[1 - pi(1/2)xexp(x(2))erfc(x)]/epsilond, where d = bl(1/2) + s and x = kappad/6(1/2). In unfolded barnase, the residual interactions lead to downward pK(a) shifts of approximately 0.33 unit, in agreement with experiment. pK(a) shifts in the unfolded state significantly affect pH dependence of protein folding stability, and the predicted effects agree very well with experimental results on barnase and four other proteins. For T4 lysozyme, the charge reversal mutation K147E is found to stabilize the unfolded state even more than the folded state (1.39 vs. 0.46 kcal/mol), leading to the experimentally observed result that the mutation is net destabilizing for the folding. The Gaussian-chain model provides a quantitative characterization of the unfolded state and may prove valuable for elucidating the energetic contributions to the stability of thermophilic proteins and the energy landscape of protein folding. PMID- 11891296 TI - Expression of CD40 identifies a unique pathogenic T cell population in type 1 diabetes. AB - Juvenile diabetes (type 1) is an autoimmune disease in which CD4(+) T cells play a major role in pathogenesis characterized by insulitis and beta cell destruction leading to clinical hyperglycemia. To date, no marker for autoimmune T cells has been described, although it was previously demonstrated that autoimmune mice have a large population of CD4(+) cells that express CD40. We show here that established, diabetogenic T cell clones of either the Th1 or Th2 phenotype are CD40-positive, whereas nondiabetogenic clones are CD40-negative. CD40 functionally signals T cell clones, inducing rapid activation of the transcription factor NFkappaB. We show that autoimmune diabetes-prone nonobese diabetic mice have high levels of CD40(+)CD4(+) T cells in the thymus, spleen, and importantly, in the pancreas. Finally, as demonstrated by adoptive transfers, CD4(+)CD40(+) cells infiltrate the pancreatic islets causing beta-cell degranulation and ultimately diabetes. PMID- 11891297 TI - Evidence for in vivo modulation of chloroplast RNA stability by 3'-UTR homopolymeric tails in Chlamydomonas reinhardtii. AB - Polyadenylation of synthetic RNAs stimulates rapid degradation in vitro by using either Chlamydomonas or spinach chloroplast extracts. Here, we used Chlamydomonas chloroplast transformation to test the effects of mRNA homopolymer tails in vivo, with either the endogenous atpB gene or a version of green fluorescent protein developed for chloroplast expression as reporters. Strains were created in which, after transcription of atpB or gfp, RNase P cleavage occurred upstream of an ectopic tRNA(Glu) moiety, thereby exposing A(28), U(25)A(3), [A+U](26), or A(3) tails. Analysis of these strains showed that, as expected, polyadenylated transcripts failed to accumulate, with RNA being undetectable either by filter hybridization or reverse transcriptase-PCR. In accordance, neither the ATPase beta-subunit nor green fluorescent protein could be detected. However, a U(25)A(3) tail also strongly reduced RNA accumulation relative to a control, whereas the [A+U] tail did not, which is suggestive of a degradation mechanism that does not specifically recognize poly(A), or that multiple mechanisms exist. With an A(3) tail, RNA levels decreased relative to a control with no added tail, but some RNA and protein accumulation was observed. We took advantage of the fact that the strain carrying a modified atpB gene producing an A(28) tail is an obligate heterotroph to obtain photoautotrophic revertants. Each revertant exhibited restored atpB mRNA accumulation and translation, and seemed to act by preventing poly(A) tail exposure. This suggests that the poly(A) tail is only recognized as an instability determinant when exposed at the 3' end of a message. PMID- 11891298 TI - Targeted new membrane addition in the cleavage furrow is a late, separate event in cytokinesis. AB - Cytokinesis in animal cells is accomplished in part by an actomyosin contractile ring. Recent work on amphibian, Drosophila, and Caenorhabditis elegans embryos implicates membrane trafficking and delivery as essential for cytokinesis. However, the relative contributions of contractile ring constriction versus membrane insertion to cytokinesis and the temporal relationship between these processes are largely unexplored. Here we monitor secretion of the extracellular matrix protein, hyalin, as a marker for new plasma membrane addition in dividing sea urchin zygotes. We find that new membrane addition occurs specifically in the cleavage furrow late in telophase independent of contractile ring constriction. The directed equatorial deposition of new furrow membrane requires astral microtubules and release of internal stores of Ca(2+), but not the presence of a central spindle. Further, cells arrested in M phase do not secrete hyalin, suggesting that mitotic exit is required for new membrane addition. These results demonstrate that astral overlap in equilaterally dividing cells not only serves to specify positioning and contraction of the contractile ring, but also to direct the delivery of new membrane to the furrow as a late, independent event during cytokinesis. PMID- 11891299 TI - Comprehensive analysis of CpG islands in human chromosomes 21 and 22. AB - CpG islands are useful markers for genes in organisms containing 5-methylcytosine in their genomes. In addition, CpG islands located in the promoter regions of genes can play important roles in gene silencing during processes such as X chromosome inactivation, imprinting, and silencing of intragenomic parasites. The generally accepted definition of what constitutes a CpG island was proposed in 1987 by Gardiner-Garden and Frommer [Gardiner-Garden, M. & Frommer, M. (1987) J. Mol. Biol. 196, 261-282] as being a 200-bp stretch of DNA with a C+G content of 50% and an observed CpG/expected CpG in excess of 0.6. Any definition of a CpG island is somewhat arbitrary, and this one, which was derived before the sequencing of mammalian genomes, will include many sequences that are not necessarily associated with controlling regions of genes but rather are associated with intragenomic parasites. We have therefore used the complete genomic sequences of human chromosomes 21 and 22 to examine the properties of CpG islands in different sequence classes by using a search algorithm that we have developed. Regions of DNA of greater than 500 bp with a G+C equal to or greater than 55% and observed CpG/expected CpG of 0.65 were more likely to be associated with the 5' regions of genes and this definition excluded most Alu-repetitive elements. We also used genome sequences to show strong CpG suppression in the human genome and slight suppression in Drosophila melanogaster and Saccharomyces cerevisiae. This finding is compatible with the recent detection of 5 methylcytosine in Drosophila, and might suggest that S. cerevisiae has, or once had, CpG methylation. PMID- 11891300 TI - Intercellular trafficking of a KNOTTED1 green fluorescent protein fusion in the leaf and shoot meristem of Arabidopsis. AB - Dominant mutations in the maize homeobox gene knotted1 (kn1) act nonautonomously during maize leaf development, indicating that Kn1 is involved in the generation or transmission of a developmental signal that passes from the inner layers of the leaf to epidermal cells. We previously found that this nonautonomous activity is correlated with the presence of KN1 protein in leaf epidermal cells, where KN1 mRNA could not be detected. Furthermore, KN1 protein expressed in Escherichia coli and labeled with a fluorescent dye can traffic between leaf mesophyll cells in microinjection assays. Here we show that green fluorescent protein (GFP) tagged KN1 is able to traffic between epidermal cells of Arabidopsis and onion. When expressed in vivo, the GFP approximately KN1 fusion trafficked from internal tissues of the leaf to the epidermis, providing the first direct evidence, to our knowledge, that KN1 can traffic across different tissue layers in the leaf. Control GFP fusions did not show this intercellular trafficking ability. GFP approximately KN1 also trafficked in the shoot apical meristem, suggesting that cell-to-cell trafficking of KN1 may be involved in its normal function in meristem initiation and maintenance. PMID- 11891301 TI - The Arf tumor suppressor gene promotes hyaloid vascular regression during mouse eye development. AB - A key tumor suppressor mechanism that is disrupted frequently in human cancer involves the ARF and p53 genes. In mouse fibroblasts, the Arf gene product responds to abnormal mitogenic signals to activate p53 and trigger either cell cycle arrest or apoptosis. Recent evidence indicates that Arf also has p53 independent functions that may contribute to its tumor suppressor activity. Using Arf(-/-) and p53(-/-) mice, we have discovered a p53-independent requirement for Arf in the developmental regression of the hyaloid vascular system (HVS) in the mouse eye. Arf is expressed in the vitreous of the eye and is induced before HVS regression in the first postnatal week. In the absence of Arf, failed HVS regression causes a pathological process that resembles persistent hyperplastic primary vitreous, a developmental human eye disease thought to have a genetic basis. These findings demonstrate an essential and unexpected role for Arf during mouse eye development, provide insights into the potential genetic basis for persistent hyperplastic primary vitreous, and indicate that Arf regulates vascular regression in a p53-independent manner. The latter finding raises the possibility that Arf may function as a tumor suppressor at least in part by regulating tumor angiogenesis. PMID- 11891302 TI - Thymocyte depletion in Trypanosoma cruzi infection is mediated by trans-sialidase induced apoptosis on nurse cells complex. AB - Trypanosoma cruzi, the causative agent of Chagas' disease, induces transient thymic aplasia early after infection-a phenomenon that still lacks a molecular explanation. The parasite sheds an enzyme known as trans-sialidase (TS), which is able to direct transfer-sialyl residues among macromolecules. Because cell surface sialylation is known to play a central role in the immune system, we tested whether the bloodstream-borne TS is responsible for the thymic alterations recorded during infection. We found that recombinant TS administered to naive mice was able to induce cell-count reduction mediated by apoptosis, mimicking cell subsets distribution and histologic findings observed during the acute phase of the infection. Thymocytes taken after TS treatment showed low response to Con A, although full ability to respond to IL-2 or IL-2 plus Con A was conserved, which resembles findings from infected animals. Alterations were found to revert several days after TS treatment. The administration of TS-neutralizing Abs to T. cruzi-infected mice prevented thymus alterations. Results indicate that the primary target for the TS-induced apoptosis is the so-called "nurse cell complex". Therefore, we report here supporting evidence that TS is the virulence factor from T. cruzi responsible for the thymic alterations via apoptosis induction on the nurse cell complex, and that TS-neutralizing Abs elicitation during infection is associated with the reversion to thymic normal parameters. PMID- 11891303 TI - Lipid mediator-induced expression of bactericidal/ permeability-increasing protein (BPI) in human mucosal epithelia. AB - Epithelial cells which line mucosal surfaces are the first line of defense against bacterial invasion and infection. Recent studies have also indicated that epithelial cells contribute significantly to the orchestration of ongoing inflammatory processes. Here, we demonstrate that human epithelial cells express bactericidal/permeability-increasing protein (BPI), an antibacterial and endotoxin-neutralizing molecule previously associated with neutrophils. Moreover, we demonstrate that such BPI expression is transcriptionally regulated by analogs of endogenously occurring anti-inflammatory eicosanoids (aspirin-triggered lipoxins, ATLa). Initial studies to verify microarray analysis revealed that epithelial cells of wide origin (oral, pulmonary, and gastrointestinal mucosa) express BPI and each is similarly regulated by aspirin-triggered lipoxins. Studies aimed at localization of BPI revealed that such expression occurs on the cell surface of cultured epithelial cell lines and dominantly localizes to epithelia in human mucosal tissue. Functional studies employing a BPI neutralizing anti-serum revealed that surface BPI blocks endotoxin-mediated signaling in epithelia and kills Salmonella typhimurium. These studies identify a previously unappreciated "molecular shield" for protection of mucosal surfaces against Gram-negative bacteria and their endotoxin. PMID- 11891304 TI - A new evolutionary scenario for the Mycobacterium tuberculosis complex. AB - The distribution of 20 variable regions resulting from insertion-deletion events in the genomes of the tubercle bacilli has been evaluated in a total of 100 strains of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canettii, Mycobacterium microti, and Mycobacterium bovis. This approach showed that the majority of these polymorphisms did not occur independently in the different strains of the M. tuberculosis complex but, rather, resulted from ancient, irreversible genetic events in common progenitor strains. Based on the presence or absence of an M. tuberculosis specific deletion (TbD1), M. tuberculosis strains can be divided into ancestral and "modern" strains, the latter comprising representatives of major epidemics like the Beijing, Haarlem, and African M. tuberculosis clusters. Furthermore, successive loss of DNA, reflected by region of difference 9 and other subsequent deletions, was identified for an evolutionary lineage represented by M. africanum, M. microti, and M. bovis that diverged from the progenitor of the present M. tuberculosis strains before TbD1 occurred. These findings contradict the often-presented hypothesis that M. tuberculosis, the etiological agent of human tuberculosis evolved from M. bovis, the agent of bovine disease. M. canettii and ancestral M. tuberculosis strains lack none of these deleted regions, and, therefore, seem to be direct descendants of tubercle bacilli that existed before the M. africanum- >M. bovis lineage separated from the M. tuberculosis lineage. This observation suggests that the common ancestor of the tubercle bacilli resembled M. tuberculosis or M. canettii and could well have been a human pathogen already. PMID- 11891305 TI - Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin. AB - Environmental stimuli such as UV, pathogen attack, and gravity can induce rapid changes in hydrogen peroxide (H(2)O(2)) levels, leading to a variety of physiological responses in plants. Catalase, which is involved in the degradation of H(2)O(2) into water and oxygen, is the major H(2)O(2)-scavenging enzyme in all aerobic organisms. A close interaction exists between intracellular H(2)O(2) and cytosolic calcium in response to biotic and abiotic stresses. Studies indicate that an increase in cytosolic calcium boosts the generation of H(2)O(2). Here we report that calmodulin (CaM), a ubiquitous calcium-binding protein, binds to and activates some plant catalases in the presence of calcium, but calcium/CaM does not have any effect on bacterial, fungal, bovine, or human catalase. These results document that calcium/CaM can down-regulate H(2)O(2) levels in plants by stimulating the catalytic activity of plant catalase. Furthermore, these results provide evidence indicating that calcium has dual functions in regulating H(2)O(2) homeostasis, which in turn influences redox signaling in response to environmental signals in plants. PMID- 11891306 TI - Supramolecular chemistry of dendrimers with functional cores. AB - Dendritic microenvironments are analogous to local environments created within protein superstructures. Correspondingly, properties of functional cores such as molecular recognition and catalytic activity are profoundly influenced by the surrounding dendritic branches. PMID- 11891307 TI - Overexpression of a Na+/H+ antiporter confers salt tolerance on a freshwater cyanobacterium, making it capable of growth in sea water. AB - The salt tolerance of a freshwater cyanobacterium, Synechococcus sp. PCC 7942, transformed with genes involved in the synthesis of a Na(+)/H(+) antiporter, betaine, catalase, and a chaperone was examined. Compared with the expression of betaine, catalase, and the chaperone, the expression of the Na(+)/H(+) antiporter gene from a halotolerant cyanobacterium (ApNhaP) drastically improved the salt tolerance of the freshwater cyanobacterium. The Synechococcus cells expressing ApNhaP could grow in BG11 medium containing 0.5 M NaCl as well as in sea water, whereas those expressing betaine, catalase, and the chaperone could not grow under those conditions. The coexpression of ApNhaP with catalase or ApNhaP with catalase and betaine did not further enhance the salt tolerance of Synechococcus cells expressing ApNhaP alone when grown in BG11 medium containing 0.5 M NaCl. Interestingly, the coexpression of ApNhaP with catalase resulted in enhanced salt tolerance of cells grown in sea water. These results demonstrate a key role of sodium ion exclusion by the Na(+)/H(+) antiporter for the salt tolerance of photosynthetic organisms. PMID- 11891308 TI - Some properties of human neuronal alpha 7 nicotinic acetylcholine receptors fused to the green fluorescent protein. AB - The functional properties and cellular localization of the human neuronal alpha7 nicotinic acetylcholine (AcCho) receptor (alpha7 AcChoR) and its L248T mutated (mut) form were investigated by expressing them alone or as gene fusions with the enhanced version of the green fluorescent protein (GFP). Xenopus oocytes injected with wild-type (wt), mutalpha7, or the chimeric subunit cDNAs expressed receptors that gated membrane currents when exposed to AcCho. As already known, AcCho currents generated by wtalpha7 receptors decay much faster than those elicited by the mutalpha7 receptors. Unexpectedly, the fusion of GFP to the wt and mutated alpha7 receptors led to opposite results: the AcCho-current decay of the wt receptors became slower, whereas that of the mutated receptors was accelerated. Furthermore, repetitive applications of AcCho led to a considerable "run-down" of the AcCho currents generated by mutalpha7-GFP receptors, whereas those of the wtalpha7-GFP receptors remained stable or increased in amplitude. The AcCho current run-down of mutalpha7-GFP oocytes was accompanied by a marked decrease of alpha-bungarotoxin binding activity. Fluorescence, caused by the chimeric receptors expressed, was seen over the whole oocyte surface but was more intense and abundant in the animal hemisphere, whereas it was much weaker in the vegetal hemisphere. We conclude that fusion of GFP to wtalpha7 and mutalpha7 receptors provides powerful tools to study the distribution and function of alpha7 receptors. We also conclude that fused genes do not necessarily recapitulate all of the properties of the original receptors. This fact must be borne close in mind whenever reporter genes are attached to proteins. PMID- 11891309 TI - The single-channel properties of human acetylcholine alpha 7 receptors are altered by fusing alpha 7 to the green fluorescent protein. AB - Neuronal nicotinic acetylcholine (AcCho) receptors composed of alpha7-subunits (alpha7-AcChoRs) are involved in many physiological activities. Nevertheless, very little is known about their single-channel characteristics. By using outside out patch-clamp recordings from Xenopus oocytes expressing wild-type (wt) alpha7 AcChoRs, we identified two classes of channel conductance: a low conductance (gamma(L)) of 72 pS and a high one (gamma(H)) of 87 pS, with mean open-times (tau(op)) of 0.6 ms. The same classes of conductances, but longer tau(op) (3 ms), were seen in experiments with chimeric alpha7 receptors in which the wtalpha7 extracellular C terminus was fused to the green fluorescent protein (wtalpha7-GFP AcChoRs). In contrast, channels with three different conductances were gated by AcCho in oocytes expressing alpha7 receptors carrying a Leu-to-Thr 248 mutation (mutalpha7) or oocytes expressing chimeric mutalpha7-GFP receptors. These conductance levels were significantly smaller, and their mean open-times were larger, than those of wtalpha7-AcChoRs. Interestingly, in the absence of AcCho, these oocytes showed single-channel openings of the same conductances, but shorter tau(op), than those activated by AcCho. Accordingly, human homomeric wtalpha7 receptors open channels of high conductance and brief lifetime, and fusion to GFP lengthens their lifetime. In contrast, mutalpha7 receptors open channels of lower conductance and longer lifetime than those gated by wtalpha7 AcChoRs, and these parameters are not greatly altered by fusing the mutalpha7 to GFP. All this evidence shows that GFP-tagging can alter importantly receptor kinetics, a fact that has to be taken into account whenever tagged proteins are used to study their function. PMID- 11891310 TI - Structural studies of the scrapie prion protein by electron crystallography. AB - Because the insolubility of the scrapie prion protein (PrP(Sc)) has frustrated structural studies by x-ray crystallography or NMR spectroscopy, we used electron crystallography to characterize the structure of two infectious variants of the prion protein. Isomorphous two-dimensional crystals of the N-terminally truncated PrP(Sc) (PrP 27-30) and a miniprion (PrP(Sc)106) were identified by negative stain electron microscopy. Image processing allowed the extraction of limited structural information to 7 A resolution. By comparing projection maps of PrP 27 30 and PrP(Sc)106, we visualized the 36-residue internal deletion of the miniprion and localized the N-linked sugars. The dimensions of the monomer and the locations of the deleted segment and sugars were used as constraints in the construction of models for PrP(Sc). Only models featuring parallel beta-helices as the key element could satisfy the constraints. These low-resolution projection maps and models have implications for understanding prion propagation and the pathogenesis of neurodegeneration. PMID- 11891312 TI - Target-induced formation of neuraminidase inhibitors from in vitro virtual combinatorial libraries. AB - Neuraminidase, a key enzyme responsible for influenza virus propagation, has been used as a template for selective synthesis of small subsets of its own inhibitors from theoretically highly diverse dynamic combinatorial libraries. We show that the library building blocks, aldehydes and amines, form significant amounts of the library components resulting from their coupling by reductive amination only in the presence of the enzyme. The target amplifies the best hits at least 120 fold. The dynamic libraries synthesized and screened in such an in vitro virtual mode form the components that possess high inhibitory activity, as confirmed by enzyme assays with independently synthesized individual compounds. PMID- 11891311 TI - Engineering a terbium-binding site into an integral membrane protein for luminescence energy transfer. AB - Luminescence resonance energy transfer with a lanthanide like Tb(3+) as donor is a useful technique for estimating intra- and intermolecular distances in macromolecules. However, the technique usually requires the use of a bulky chelator with a flexible linker attached to a Cys residue to bind Tb(3+) and, for intramolecular studies, an acceptor fluorophor attached to another Cys residue in the same protein. Here, an engineered EF- hand motif is incorporated into the central cytoplasmic loop of the lactose permease of Escherichia coli generating a high-affinity site for Tb(3+) (K(Tb)(3+) approximately 4.5 microM) or Gd(3+) (K(Gd)(3+) approximately 2.3 microM). By exciting a Trp residue in the coordination sequence, Tb(3+) bound to the EF-hand motif is sensitized specifically, and the efficiency of energy transfer to strategically placed Cys residues labeled with fluorophors is measured. In this study, we use the technique to measure distance from the EF-hand in the central cytoplasmic loop of lactose permease to positions 179 or 169 at the center or periplasmic end of helix VI, respectively. The average calculated distances of approximately 23 A (position 179) and approximately 33 A (position 169) observed with three different fluorophors as acceptors agree well with the geometry of a slightly tilted alpha-helix. The approach should be of general use for studying static and dynamic aspects of polytopic membrane protein structure and function. PMID- 11891313 TI - Stop codons preceded by rare arginine codons are efficient determinants of SsrA tagging in Escherichia coli. AB - The SsrA or tmRNA quality control system intervenes when ribosomes stall on mRNAs and directs the addition of a C-terminal peptide tag that targets the modified polypeptide for degradation. Although hundreds of SsrA-tagged proteins can be detected in cells when degradation is prevented, most of these species have not been identified. Consequently, the mRNA sequence determinants that cause ribosome stalling and SsrA tagging are poorly understood. SsrA tagging of Escherichia coli ribokinase occurs at three specific sites at or near the C terminus of this protein. The sites of tagging correspond to ribosome stalling at the termination codon and at rare AGG codons encoding Arg-307 and Arg-309, the antepenultimate and C-terminal residues of E. coli ribokinase. Mutational analyses and studies of the effects of overexpressing the tRNA that decodes AGG reveal that the combination of a rare arginine codon at the C terminus and the adjacent inefficient UGA termination codon act to recruit the SsrA-tagging system, presumably by slowing the rate of translation elongation and termination. PMID- 11891314 TI - Chemical synthesis gets a fillip from molecular recognition and self-assembly processes. AB - By drawing on nature conceptually, the practice of chemical synthesis can now be broadened in scope to include the supramolecular synthesis of supermolecules and their postassembly covalent modification to form mechanically interlocked molecules, which the chemist can use to build functioning nanosystems after the style of those found in the living world. PMID- 11891316 TI - The gas phase reaction of singlet dioxygen with water: a water-catalyzed mechanism. AB - Stimulated by the recent surprising results from Wentworth et al. [Wentworth, A. D., Jones, L. H., Wentworth, P., Janda, K. D. & Lerner, R. A. (2000) Proc. Natl. Acad. Sci. USA 97, 10930-10935] that Abs efficiently catalyze the conversion of molecular singlet oxygen ((1)O(2)) plus water to hydrogen peroxide (HOOH), we used quantum chemical methods (B3LYP density functional theory) to delineate the most plausible mechanisms for the observed efficient conversion of water to HOOH. We find two reasonable pathways. In Pathway I, (i) H(2)O catalyzes the reaction of (1)O(2) with a second water to form HOOOH; (ii) two HOOOH form a dimer, which rearranges to form the HOO-HOOO + H(2)O complex; (iii) HOO-HOOO rearranges to HOOH-OOO, which subsequently reacts with H(2)O to form H(2)O(4) + HOOH; and (iv) H(2)O(4) rearranges to the cyclic dimer (HO(2))(2), which in turn forms HOOH plus (1)O(2) or (3)O(2). Pathway II differs in that step ii is replaced with the reaction between HOOOH and (1)O(2), leading to the formation of HOO-HOOO. This then proceeds to similar products. For a system with (18)O H(2)O, Pathway I leads to a 2.2:1 ratio of (16)O:(18)O in the product HOOH, whereas Pathway II leads to 3:1. These ratios are in good agreement with the 2.2:1 ratio observed in isotope experiments by Wentworth et al. These mechanisms lead to two HOOH per initial (1)O(2) or one, depending on whether the product of step iv is (1)O(2) or (3)O(2), in good agreement with the experimental result of 2.0. In addition to the Ab-induced reactions, the hydrogen polyoxides (H(2)O(3) and H(2)O(4)) formed in these mechanisms and their decomposition product polyoxide radicals (HO(2), HO(3)) may play a role in combustion, explosions, atmospheric chemistry, and the radiation chemistry in aqueous systems. PMID- 11891315 TI - Disrupting the IL-4 gene rescues mice homozygous for the tight-skin mutation from embryonic death and diminishes TGF-beta production by fibroblasts. AB - The TSK/TSK mutation is embryonic lethal; embryos have been reported to die at 7 8 days of gestational age. Crossing TSK/+, IL-4+/- mice revealed that disrupting one or both IL-4 alleles allowed survival of 29 and 47%, respectively, of TSK/TSK mice. These mice failed to develop cutaneous hyperplasia but did exhibit the emphysema that is found in TSK/+ mice. We showed that IL-4 stimulation of fibroblasts increased the level of transforming growth factor-beta (TGF-beta) mRNA and that lungs of TSK/+, IL-4-/- mice had substantially less TGF-beta mRNA than lungs of TSK/+, IL-4+/+ mice. Thus IL-4 seems to regulate the expression of TGF-beta in fibroblasts, providing an explanation for the absence of cutaneous hyperplasia in TSK/+, IL-4Ralpha-/- and TSK/+, TGF-beta+/- mice. PMID- 11891317 TI - Bombesin antagonists inhibit growth of MDA-MB-435 estrogen-independent breast cancers and decrease the expression of the ErbB-2/HER-2 oncoprotein and c-jun and c-fos oncogenes. AB - Previous studies showed that antagonists of bombesin (BN)/gastrin-releasing peptide (GRP) inhibit the growth of various cancers by interfering with the growth-stimulatory effects of BN-like peptides and down-regulating epidermal growth factor receptors on tumors. Because the overexpression of the human epidermal growth factor receptor-2 (ErbB-2/HER-2/neu) oncogene plays a role in the progression of many breast cancers, we investigated whether BN/GRP antagonists can affect HER-2 in mammary tumors. Female nude mice bearing orthotopic xenografts of MDA-MB-435 human estrogen-independent breast cancers were treated daily with BN/GRP antagonists RC-3095 (20 microg) or RC-3940-II (10 microg) for 6 weeks. The expression of BN/GRP receptors on tumors was analyzed by reverse transcription-PCR and immunoblotting. We also evaluated whether the mRNA expression for the c-jun and c-fos oncogenes is affected by the therapy. Both BN/GRP antagonists significantly inhibited growth of MDA-MB-435 cancers; RC-3095 reduced tumor volume by 40% and RC-3940-II by 65%. The GRP receptors (subtype 1) were detected in MDA-MB-435 tumors, showing that they mediate the inhibitory effect of the antagonists. Tumor inhibition was associated with a substantial reduction in the expression of mRNA and protein levels of the ErbB/HER receptor family as well as with a decrease in the expression of c-jun and c-fos oncogenes. BN/GRP antagonists RC-3940-II and RC-3095 could be considered for endocrine therapy of estrogen-independent breast cancers that express members of the ErbB/HER receptor family and the c-jun and c-fos oncogenes. PMID- 11891318 TI - Linkage disequilibrium between the beta frequency of the human EEG and a GABAA receptor gene locus. AB - Human brain oscillations represent important features of information processing and are highly heritable. A common feature of beta oscillations (13-28 Hz) is the critical involvement of networks of inhibitory interneurons as pacemakers, gated by gamma-aminobutyric acid type A (GABA(A)) action. Advances in molecular and statistical genetics permit examination of quantitative traits such as the beta frequency of the human electroencephalogram in conjunction with DNA markers. We report a significant linkage and linkage disequilibrium between beta frequency and a set of GABA(A) receptor genes. Uncovering the genes influencing brain oscillations provides a better understanding of the neural function involved in information processing. PMID- 11891319 TI - Restoration of fragile histidine triad (FHIT) expression induces apoptosis and suppresses tumorigenicity in lung and cervical cancer cell lines. AB - Loss of expression of the Fhit protein is often associated with the development of many human epithelial cancers, including lung and cervical carcinomas. Restoration of Fhit expression in cell lines derived from these tumors has however yielded conflicting results, prompting the need for careful evaluation of the oncosuppressive potential of FHIT. In the present study, we have investigated the effect of Fhit reintroduction in seven lung cancer and three cervical cancer cell lines. To achieve efficient gene transfer and high levels of transgene expression, we have used an adenoviral vector to transduce the FHIT gene. The induction of apoptosis was evaluated by using the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay and propidium iodide staining. Activation of caspases was detected by using Western blot analysis, and tumorigenic potential of transduced cells in the nude mouse was also assessed. Restoration of Fhit expression induced apoptosis in all Fhit negative cell lines, with Calu-1, H460, and A549 being the most susceptible among the lung cancer cell lines and SiHa cells among cervical carcinomas. Activation of caspase-8 was always associated with Fhit-mediated apoptosis, and in vivo tumorigenicity was either abolished by FHIT gene transfer (in H460 and SK-Mes cells) or strongly suppressed (in A549 and SiHa cells). Our data demonstrate oncosuppressive properties and strong proapoptotic activity of the Fhit protein in lung and cervical cancer cell lines and strengthens the hypothesis of its possible use as a therapeutic tool. PMID- 11891320 TI - Design of an HIV-1 lentiviral-based gene-trap vector to detect developmentally regulated genes in mammalian cells. AB - The recent development of HIV-1 lentiviral vectors is especially useful for gene transfer because they achieve efficient integration into nondividing cell genomes and successful long-term expression of the transgene. These attributes make the vector useful for gene delivery, mutagenesis, and other applications in mammalian systems. Here we describe two HIV-1-based lentiviral vector derivatives, pZR-1 and pZR-2, that can be used in gene-trap experiments in mammalian cells in vitro and in vivo. Each lentiviral gene-trap vector contains a reporter gene, either beta-lactamase or enhanced green fluorescent protein (EGFP), that is inserted into the U3 region of the 3' long terminal repeat. Both of the trap vectors readily integrate into the host genome by using a convenient infection technique. Appropriate insertion of the vector into genes causes EGFP or beta-lactamase expression. This technique should facilitate the rapid enrichment and cloning of the trapped cells and provides an opportunity to select subpopulations of trapped cells based on the subcellular localization of reporter genes. Our findings suggest that the reporter gene is driven by an upstream, cell-specific promoter during cell culture and cell differentiation, which further supports the usefulness of lentivirus-based gene-trap vectors. Lentiviral gene-trap vectors appear to offer a wealth of possibilities for the study of cell differentiation and lineage commitment, as well as for the discovery of new genes. PMID- 11891321 TI - Cells exposed to antifolates show increased cellular levels of proteins fused to dihydrofolate reductase: a method to modulate gene expression. AB - Human cells exposed to antifolates show a rapid increase in the levels of the enzyme dihydrofolate reductase (DHFR). We hypothesized that this adaptive response mechanism can be used to elevate cellular levels of proteins fused to DHFR. In this study, mouse cells transfected to express a green fluorescent protein-DHFR fusion protein and subsequently exposed to the antifolate trimetrexate (TMTX) showed a specific and time-dependent increase in cellular levels of the fusion protein. Next, human HCT-8 and HCT-116 colon cancer cells retrovirally transduced to express a DHFR-herpes simplex virus 1 thymidine kinase (HSV1 TK) fusion protein and treated with the DHFR inhibitor TMTX exhibited increased levels of the DHFR-HSV1 TK fusion protein and an increase in ganciclovir sensitivity by 250-fold. The level of fusion protein in antifolate treated human tumor cells was increased in response to a 24-h exposure of methotrexate, trimetrexate, as well as dihydrofolate. This effect depended on the antifolate concentration and was independent of the fusion-protein mRNA levels, consistent with this increase occurring at a translational level. In a xenograft model, nude rats bearing DHFR-HSV1 TK-transduced HCT-8 tumors and treated with TMTX showed, after 24 h, a 2- to 4-fold increase of fusion-protein levels in tumor tissue from treated animals compared with controls, as determined by Western blotting. The fusion-protein increase was imaged with positron-emission tomography, where a substantially enhanced signal of the transduced tumor was detected in animals after antifolate administration. Drug-mediated elevation of cellular DHFR-fused proteins is a very useful method to modulate gene expression in vivo for imaging as well as therapeutic purposes. PMID- 11891322 TI - Small-molecule antagonists of Myc/Max dimerization inhibit Myc-induced transformation of chicken embryo fibroblasts. AB - Myc is a transcriptional regulator of the basic helix-loop-helix leucine zipper protein family. It has strong oncogenic potential, mutated or virally transduced forms of Myc induce lymphoid tumors in animals, and deregulated expression of Myc is associated with numerous types of human cancers. For its oncogenic activity, Myc must dimerize with the ubiquitously expressed basic helix-loop-helix leucine zipper protein Max. This requirement for dimerization may allow control of Myc activity with small molecules that interfere with Myc/Max dimerization. We have measured Myc/Max dimerization with fluorescence resonance energy transfer and have screened combinatorial chemical libraries for inhibitors of dimerization. Candidate inhibitors were isolated from a peptidomimetics library. Inhibition of Myc/Max interaction was validated by ELISA and electrophoretic mobility-shift assay. Two of the candidate inhibitors also interfere with Myc-induced oncogenic transformation in chicken embryo fibroblast cultures. Our work provides proof of principle for the identification of small molecule inhibitors of protein-protein interactions by using high-throughput screens of combinatorial chemical libraries. PMID- 11891323 TI - Quantitative measurement of translesion replication in human cells: evidence for bypass of abasic sites by a replicative DNA polymerase. AB - Mutations in oncogenes and tumor suppressor genes are critical in the development of cancer. A major pathway for the formation of mutations is the replication of unrepaired DNA lesions. To better understand the mechanism of translesion replication (TLR) in mammals, a quantitative assay for TLR in cultured cells was developed. The assay is based on the transient transfection of cultured cells with a gapped plasmid, carrying a site-specific lesion in the gap region. Filling in of the gap by TLR is assayed in a subsequent bioassay, by the ability of the plasmid extracted from the cells, to transform an Escherichia coli indicator strain. Using this method it was found that TLR through a synthetic abasic site in the adenocarcinoma H1299, the osteogenic sarcoma Saos-2, the prostate carcinoma PC3, and the hepatoma Hep3B cell lines occurred with efficiencies of 92 +/- 6%, 32 +/- 2%, 72 +/- 4%, and 26 +/- 3%, respectively. DNA sequence analysis showed that 85% of the bypass events in H1299 cells involved insertion of dAMP opposite the synthetic abasic site. Addition of aphidicolin, an inhibitor of DNA polymerases alpha, delta, and epsilon, caused a 4.4-fold inhibition of bypass. Analysis of two XP-V cell lines, defective in DNA polymerase eta, showed bypass of 89%, indicating that polymerase eta is not essential for bypass of abasic sites. These results suggest that in human cells bypass of abasic sites does not require the bypass-specific DNA polymerase eta, but it does require at least one of the replicative DNA polymerases, alpha, delta, or epsilon. The quantitative TLR assay is expected to be useful in the molecular analysis of lesion bypass in a large variety of cultured mammalian cells. PMID- 11891324 TI - The embryonic stem cell transcription factors Oct-4 and FoxD3 interact to regulate endodermal-specific promoter expression. AB - The POU homeodomain protein Oct-4 and the Forkhead Box protein FoxD3 (previously Genesis) are transcriptional regulators expressed in embryonic stem cells. Down regulation of Oct-4 during gastrulation is essential for proper endoderm development. After gastrulation, FoxD3 is generally down-regulated during early endoderm formation, although it specifically remains expressed in the embryonic neural crest. In these studies, we have found that Oct-4 and FoxD3 can bind to identical regulatory DNA sequences. In addition, Oct-4 physically interacted with the FoxD3 DNA-binding domain. Cotransfection of Oct-4 and FoxD3 expression vectors activated the osteopontin enhancer, which is expressed in totipotent embryonic stem cells. FoxA1 and FoxA2 (previously HNF-3alpha and HNF-3beta) are Forkhead Box transcription factors that participate in liver and lung formation from foregut endoderm. Although FoxD3 activated the FoxA1 and FoxA2 promoters, Oct-4 inhibited FoxD3 activation of the FoxA1 and FoxA2 endodermal promoters. These data indicate that Oct-4 functions as a corepressor of FoxD3 to provide embryonic lineage-specific transcriptional regulatory activity to maintain appropriate developmental timing. PMID- 11891325 TI - Social intelligence, innovation, and enhanced brain size in primates. AB - Despite considerable current interest in the evolution of intelligence, the intuitively appealing notion that brain volume and "intelligence" are linked remains untested. Here, we use ecologically relevant measures of cognitive ability, the reported incidence of behavioral innovation, social learning, and tool use, to show that brain size and cognitive capacity are indeed correlated. A comparative analysis of 533 instances of innovation, 445 observations of social learning, and 607 episodes of tool use established that social learning, innovation, and tool use frequencies are positively correlated with species' relative and absolute "executive" brain volumes, after controlling for phylogeny and research effort. Moreover, innovation and social learning frequencies covary across species, in conflict with the view that there is an evolutionary tradeoff between reliance on individual experience and social cues. These findings provide an empirical link between behavioral innovation, social learning capacities, and brain size in mammals. The ability to learn from others, invent new behaviors, and use tools may have played pivotal roles in primate brain evolution. PMID- 11891326 TI - Characterization of mutant tobacco mosaic virus coat protein that interferes with virus cell-to-cell movement. AB - Expression of tobacco mosaic virus (TMV) coat protein (CP) in plants confers resistance to infection by TMV and related tobamoviruses. Certain mutants of the CP (CP(T42W)) provide much greater levels of resistance than wild-type (wt) CP. In the present work, infection induced by RNA transcripts of TMV clones that contain wt CP or mutant CP(T42W) fused to the green fluorescent protein (GFP) (TMV-CP:GFP, TMV-CP(T42W):GFP) and clones harboring TMV movement protein (MP):GFP were followed in nontransgenic and transgenic tobacco BY-2 protoplasts and Nicotiana tabaccum Xanthi-nn plants that express wt CP or CP(T42W). On nontransgenic and wt CP transgenic plants, TMV-CP:GFP produced expanding, highly fluorescent disk-shaped areas. On plants expressing CP(T42W), infection by TMV CP:GFP or TMV-MP:GFP-CP produced infection sites of smaller size that were characterized by low fluorescence, reflecting reduced levels of virus spread and reduced accumulation of both CP:GFP and MP:GFP. TMV-CP(T42W):GFP failed to produce visible infection sites on nontransgenic plants, yet produced normal infection sites on MP-transgenic plants that produce MP. TMV infection of transgenic BY-CP(T42W) protoplasts resulted in very low levels of MP accumulation, whereas on BY-CP protoplasts (containing wt CP), infection produced higher levels of MP than in nontransgenic BY-2 cells. The results suggest that wt CP has a positive effect on the production of MP, whereas the CP(T42W) has a negative effect on MP accumulation and/or function. This effect results in very high levels of resistance to TMV infection in plants containing CP(T42W). This report shows that the CP of a plant virus regulates production of the MP, and that a mutant CP interferes with MP accumulation and cell-to-cell movement of infection. PMID- 11891327 TI - Ammonium/methylammonium transport (Amt) proteins facilitate diffusion of NH3 bidirectionally. AB - The ammonium/methylammonium transport (Amt) proteins of enteric bacteria and their homologues, the methylammonium/ammonium permeases of Saccharomyces cerevisiae, are required for fast growth at very low concentrations of the uncharged species NH(3). For example, they are essential at low ammonium (NH(4)+ + NH(3)) concentrations under acidic conditions. Based on growth studies in batch culture, the Amt protein of Salmonella typhimurium (AmtB) cannot concentrate either NH(3) or NH(4)+ and this organism appears to have no means of doing so. We now show that S. typhimurium releases ammonium into the medium when grown on the alternative nitrogen source arginine and that outward diffusion of ammonium is enhanced by the activity of AmtB. The latter result indicates that AmtB acts bidirectionally. We also confirm a prediction that the AmtB protein would be required at pH 7.0 in ammonium-limited continuous culture, i.e., when the concentration of NH(3) is < or =50 nM. Together with our previous studies, current results are in accord with the view that Amt and methylammonium/ammonium permease proteins increase the rate of diffusion of the uncharged species NH(3) across the cytoplasmic membrane. These proteins are examples of protein facilitators for a gas. PMID- 11891328 TI - Combined expression of pTalpha and Notch3 in T cell leukemia identifies the requirement of preTCR for leukemogenesis. AB - Notch receptors are conserved regulators of cell fate and have been implicated in the regulation of T cell differentiation and lymphomagenesis. However, neither the generality of Notch involvement in leukemia, nor the molecules with which Notch may interact have been clarified. Recently, we showed that transgenic mice expressing the constitutively active intracellular domain of Notch3 in thymocytes and T cells developed early and aggressive T cell neoplasias. Although primarily splenic, the tumors sustained features of immature thymocytes, including expression of pTalpha, a defining component of the pre T cell receptor, known to be a potent signaling complex provoking thymocyte survival, proliferation, and activation. Thus, enforced expression of Notch3, which is ordinarily down regulated as thymocytes mature, may sustain pre T cell receptor expression, causing dysregulated hyperplasia. This hypothesis has been successfully tested in this article by the observation that deletion of pTalpha in Notch3 transgenic mice abrogates tumor development, indicating a crucial role for pTalpha in T cell leukemogenesis. Parallel observations were made in humans, in that all T cell acute lymphoblastic leukemias examined showed expression of Notch3 and of the Notch target gene HES-1, as well as of pTalpha a and b transcripts, whereas the expression of all these genes was dramatically reduced or absent in remission. Together, these results suggest that the combined expression of Notch3 and pTalpha sustains T cell leukemogenesis and may represent pathognomonic molecular features of human T-ALL. PMID- 11891329 TI - Role for FGFR2IIIb-mediated signals in controlling pancreatic endocrine progenitor cell proliferation. AB - Pancreatic development is a classic example of epithelium-mesenchyme interaction. During embryonic life, signals from the mesenchyme control the proliferation of precursor cells within the pancreatic epithelium and their differentiation into endocrine or acinar cells. It has been shown that signals from the mesenchyme activate epithelial cell proliferation but repress development of the pancreatic epithelium into endocrine cells. Here, experiments with specific inhibitors established that mesenchymal effects on epithelial cell development depended on the mitogen-activated protein kinase pathway. Then we demonstrated that these effects of the mesenchyme were mimicked by fibroblast growth factor 7 (FGF7), a specific ligand of FGFR2IIIb, which is a tyrosine kinase receptor of the FGF receptor family. When pancreatic epithelium expressing FGFR2IIIb was grown with FGF7, epithelial cell growth occurred in a concentration-dependent manner, whereas endocrine tissue development was repressed. The epithelial cells that proliferated in response to FGF7 were endocrine pancreatic precursor cells, as shown by their differentiation en masse into endocrine cells on FGF7 removal. Thus, efficient propagation of pancreatic progenitor cells can be achieved in vitro by exposure to FGF7, which does not affect their ability to differentiate en masse into endocrine cells on FGF7 removal. PMID- 11891330 TI - sgk, a primary glucocorticoid-induced gene, facilitates memory consolidation of spatial learning in rats. AB - By using differential display PCR, we have identified 98 cDNA fragments from the rat dorsal hippocampus that are expressed differentially between the fast learners and slow learners in the water maze learning task. One of these cDNA fragments encodes the rat serum- and glucocorticoid-inducible kinase (sgk) gene. Northern blot analysis revealed that the sgk mRNA level was approximately 4-fold higher in the hippocampus of fast learners than slow learners. In situ hybridization results indicated that sgk mRNA level was increased markedly in CA1, CA3, and dentate gyrus of hippocampus in fast learners. Transient transfection of the sgk mutant DNA to the CA1 area impaired, whereas transfection of the sgk wild-type DNA facilitated water maze performance in rats. These results provide direct evidence that enhanced sgk expression facilitates memory consolidation of spatial learning in rats. These results also elucidate the molecular mechanism of glucocorticoid-induced memory facilitation in mammals. PMID- 11891331 TI - Deletion of the thyroid hormone receptor alpha 1 prevents the structural alterations of the cerebellum induced by hypothyroidism. AB - Thyroid hormone (T3) controls critical aspects of cerebellar development, such as migration of postmitotic granule cells and terminal differentiation of Purkinje cells. T3 acts through nuclear receptors (TR) of two types, TRalpha1 and TRbeta, that either repress or activate gene expression. We have analyzed the cerebellar structure of developing mice lacking the TRalpha1 isoform, which normally accounts for about 80% of T3 receptors in the cerebellum. Contrary to what was expected, granule cell migration and Purkinje cell differentiation were normal in the mutant mice. Even more striking was the fact that when neonatal hypothyroidism was induced, no alterations in cerebellar structure were observed in the mutant mice, whereas the wild-type mice showed delayed granule cell migration and arrested Purkinje cell growth. The results support the idea that repression by the TRalpha1 aporeceptor, and not the lack of thyroid hormone, is responsible for the hypothyroid phenotype. This conclusion was supported by experiments with the TRbeta-selective compound GC-1. Treatment of hypothyroid animals with T3, which binds to TRalpha1 and TRbeta, prevents any defect in cerebellar structure. In contrast, treatment with GC-1, which binds to TRbeta but not TRalpha1, partially corrects Purkinje cell differentiation but has no effect on granule cell migration. Our data indicate that thyroid hormone has a permissive effect on cerebellar granule cell migration through derepression by the TRalpha1 isoform. PMID- 11891332 TI - MEKK1 is essential for cardiac hypertrophy and dysfunction induced by Gq. AB - Signaling via mitogen-activated protein kinases is implicated in heart failure induced by agonists for G protein-coupled receptors that act via the G protein Galphaq. However, this assertion relies heavily on pharmacological inhibitors and dominant-interfering proteins and not on gene deletion. Here, we show that endogenous cardiac MAPK/ERK kinase kinase-1 (MEKK1)/(MAP3K1), a mitogen-activated protein kinase kinase kinase, is activated by heart-restricted overexpression of Galphaq in mice. In cardiac myocytes derived from embryonic stem cells in culture, homozygous disruption of MEKK1 selectively impaired c-Jun N-terminal kinase activity in the absence or presence of phenlyephrine, a Galphaq-dependent agonist. Other terminal mitogen-activated protein kinases were unaffected. In mice, the absence of MEKK1 abolished the increase in cardiac mass, myocyte size, hypertrophy-associated atrial natriuretic factor induction, and c-Jun N-terminal kinase activation by Galphaq, and improved ventricular mechanical function. Thus, MEKK1 mediates cardiac hypertrophy induced by Galphaq in vivo and is a logical target for drug development in heart disease involving this pathway. PMID- 11891333 TI - Aluminum-dependent regulation of intracellular silicon in the aquatic invertebrate Lymnaea stagnalis. AB - Silicon is essential for some plants, diatoms, and sponges but, in higher animals, its endogenous regulation has not been demonstrated. Silicate ions may be natural ligands for aluminum and here we show that, in the freshwater snail (Lymnaea stagnalis), intracellular silicon seems specifically up-regulated in response to sublethal aluminum exposure. X-ray microanalysis showed that exposure of snails to low levels of aluminum led to its accumulation in lysosomal granules, accompanied by marked up-regulation of silicon. Increased lysosomal levels of silicon were a specific response to aluminum because cadmium and zinc had no such effect. Furthermore, intra-lysosomal sulfur from metallothionein and other sulfur-containing ligands was increased after exposure to cadmium and zinc but not aluminum. To ensure that these findings indicated a specific in vivo response, and not ex vivo formation of hydroxy-aluminosilicates (HAS) from added aluminum (555 microg/liter) and water-borne silicon (43 microg/liter), two further studies were undertaken. In a ligand competition assay the lability of aluminum (527 microg/liter) was completely unaffected by the presence of silicon (46 microg/liter), suggesting the absence of HAS. In addition, exogenous silicon (6.5 mg/liter), added to the water column to promote formation of HAS, caused a decrease in lysosomal aluminum accumulation, showing that uptake of HAS would not explain the loading of aluminum into lysosomal granules. These findings, and arguments on the stability, lability, and kinetics of aluminum-silicate interactions, suggest that a silicon-specific mechanism exists for the in vivo detoxification of aluminum, which provides regulatory evidence of silicon in a multicellular organism. PMID- 11891334 TI - Interaction of RNA polymerase with forked DNA: evidence for two kinetically significant intermediates on the pathway to the final complex. AB - RNA polymerase forms competitor-resistant complexes with "forked DNA" templates that are double-stranded from the -35 promoter region through the first base pair of the -10 region, with an additional unpaired A at the 3' end of the nontemplate strand. These types of substrates were introduced recently as model templates for the study of DNA-protein interactions occurring in the early stages of the formation of RNA polymerase-promoter open complexes. We have performed kinetic and equilibrium measurements of interactions of wild-type and mutant RNA polymerases bearing substitutions in the sigma(70) initiation factor, with forked DNA of wild-type and mutant sequence. Our observations reveal that formation of a competitor-resistant complex between RNA polymerase and forked DNA, similar to the formation of open complexes at promoters, is a multistep process, and some of the sequentially formed intermediates along the two pathways share common properties. This work establishes, for the forked template, progression through these intermediates in the absence of downstream DNA and validates the use of forked DNA to determine the effects of changes in promoter or RNA polymerase sequence on the process of open complex formation. PMID- 11891335 TI - Induction of p57(KIP2) expression by p73beta. AB - The p53-related protein p73 has many functions similar to that of p53 including the ability to induce cell-cycle arrest and apoptosis. Both p53 and p73 function as transcription factors, and p73 activates expression of many genes that also are regulated by p53. Despite their similarities, it is evident that p53 and p73 are not interchangeable functionally, with p73 playing a role in normal growth and development that is not shared by p53. In this paper we describe the ability of p73beta but not p53 to activate expression of the cyclin-dependent kinase inhibitor p57(KIP) and KvLQT1, two genes that are coregulated in an imprinted region of the genome. Our results suggest that p73 may regulate expression of genes through mechanisms that are not shared by p53, potentially explaining the different contributions of p53 and p73 to normal development. PMID- 11891336 TI - Protein unfolding: rigidity lost. AB - We relate the unfolding of a protein to its loss of structural stability or rigidity. Rigidity and flexibility are well defined concepts in mathematics and physics, with a body of theorems and algorithms that have been applied successfully to materials, allowing the constraints in a network to be related to its deformability. Here we simulate the weakening or dilution of the noncovalent bonds during protein unfolding, and identify the emergence of flexible regions as unfolding proceeds. The transition state is determined from the inflection point in the change in the number of independent bond-rotational degrees of freedom (floppy modes) of the protein as its mean atomic coordination decreases. The first derivative of the fraction of floppy modes as a function of mean coordination is similar to the fraction-folded curve for a protein as a function of denaturant concentration or temperature. The second derivative, a specific heat-like quantity, shows a peak around a mean coordination of = 2.41 for the 26 diverse proteins we have studied. As the protein denatures, it loses rigidity at the transition state, proceeds to a state where just the initial folding core remains stable, then becomes entirely denatured or flexible. This universal behavior for proteins of diverse architecture, including monomers and oligomers, is analogous to the rigid to floppy phase transition in network glasses. This approach provides a unifying view of the phase transitions of proteins and glasses, and identifies the mean coordination as the relevant structural variable, or reaction coordinate, along the unfolding pathway. PMID- 11891337 TI - Somatic action potentials are sufficient for late-phase LTP-related cell signaling. AB - A question of critical importance confronting neuroscientists today is how biochemical signals initiated at a synapse are conveyed to the nucleus. This problem is particularly relevant to the generation of the late phases of long term potentiation (LTP). Here we provide evidence that some signaling pathways previously associated with late-LTP can be activated in hippocampal CA1 neurons without synaptic activity; somatic action potentials, induced by backfiring the cells, were found to be sufficient for phosphorylation of extracellular signal regulated kinase-1/2 and cAMP response element-binding protein, as well as for induction of zif268. Furthermore, such antidromic stimulation was adequate to rescue "tagged" synapses (early-LTP) from decay. These results show that a synapse-to-nucleus signal is not necessary for late-phase LTP-associated signaling cascades in the regulation of gene expression. PMID- 11891338 TI - Embryonic stem cells and somatic cells differ in mutation frequency and type. AB - Pluripotent embryonic stem (ES) cells have been used to produce genetically modified mice as experimental models of human genetic diseases. Increasingly, human ES cells are being considered for their potential in the treatment of injury and disease. Here we have shown that mutation in murine ES cells, heterozygous at the selectable Aprt locus, differs from that in embryonic somatic cells. The mutation frequency in ES cells is significantly lower than that in mouse embryonic fibroblasts, which is similar to that in adult cells in vivo. The distribution of spontaneous mutagenic events is remarkably different between the two cell types. Although loss of the functional allele is the predominant mutation type in both cases, representing about 80% of all events, mitotic recombination accounted for all loss of heterozygosity events detected in somatic cells. In contrast, mitotic recombination in ES cells appeared to be suppressed and chromosome loss/reduplication, leading to uniparental disomy (UPD), represented more than half of the loss of heterozygosity events. Extended culture of ES cells led to accumulation of cells with adenine phosphoribosyltransferase deficiency and UPD. Because UPD leads to reduction to homozygosity at multiple recessive disease loci, including tumor suppressor loci, in the affected chromosome, the increased risk of tumor formation after stem cell therapy should be viewed with concern. PMID- 11891339 TI - Learning to see the trees before the forest: reversible deactivation of the superior colliculus during learning of local and global visual features. AB - Previous studies have established that deactivation of the superior colliculus severely retards the normally rapid learning of pattern discriminations in the mature cat. The purpose of this study was to test the hypothesis that the midbrain plays an important role in the learning of simple pattern discriminations and that the contribution of this pathway is to the perception of global, rather than local, features of a figure. To answer this question, pattern discrimination learning was studied in three intact cats and in three experimental cats during bilateral reversible deactivation of the superficial layers of the superior colliculus (SC). The animals concurrently learned to discriminate three pairs of compound visual patterns composed of small (local element) Ts or 7s. Congruent and incongruent stimulus pairs were large (global) Ts vs. 7s comprising the same or different local elements, respectively. The random stimulus pair consisted of randomly scattered local elements (Ts vs. 7s). The animals were trained to respond to the large Ts in the congruent and incongruent pairs and the small Ts in the random pair. In the normal cats, learning of the random pair was much slower than the learning of the congruent and incongruent pairs. This finding demonstrated the theory of global precedence, because the animals learned the global features of the congruent and incongruent pairs much more quickly than the local features of the random pair. In contrast, during bilateral deactivation of the superficial layers of the SC, the learning of the incongruent pair was significantly retarded and took longer to learn than the random pair. Congruent and random pair learning rates were unchanged. The specific deficit in learning the incongruent pair indicates that the learning of global, but not local, elements of the visual pattern is impaired during deactivation of the SC. The unimpaired use of local features permitted the animals to learn the congruent and random pairs at normal rates. Therefore, deactivation of the superficial layers of the SC during pattern discrimination learning reverses the precedence for global visual features that is typical of normal learning. PMID- 11891340 TI - Crystal structure of the OpcA integral membrane adhesin from Neisseria meningitidis. AB - OpcA is an integral outer membrane protein from Neisseria meningitidis, the causative agent of meningococcal meningitis and septicemia. It mediates the adhesion of N. meningitidis to epithelial and endothelial cells by binding to vitronectin and proteoglycan cell-surface receptors. Here, we report the determination of the crystal structure of OpcA to 2.0 A resolution. OpcA adopts a 10-stranded beta-barrel structure with extensive loop regions that protrude above the predicted surface of the membrane. The second external loop adopts an unusual conformation, traversing the axis of the beta-barrel and apparently blocking formation of a pore through the membrane. Loops 2, 3, 4, and 5 associate to form one side of a crevice in the external surface of the structure, the other side being formed by loop 1. The crevice is lined by positively charged residues and would form an ideal binding site for proteoglycan polysaccharide. The structure, therefore, suggests a model for how adhesion of this important human pathogen to proteoglycan is mediated at the molecular level. PMID- 11891341 TI - Nuclear entry and CRM1-dependent nuclear export of the Rous sarcoma virus Gag polyprotein. AB - The retroviral Gag polyprotein directs budding from the plasma membrane of infected cells. Until now, it was believed that Gag proteins of type C retroviruses, including the prototypic oncoretrovirus Rous sarcoma virus, were synthesized on cytosolic ribosomes and targeted directly to the plasma membrane. Here we reveal a previously unknown step in the subcellular trafficking of the Gag protein, that of transient nuclear localization. We have identified a targeting signal within the N-terminal matrix domain that facilitates active nuclear import of the Gag polyprotein. We also found that Gag is transported out of the nucleus through the CRM1 nuclear export pathway, based on observations that treatment of virus-expressing cells with leptomycin B resulted in the redistribution of Gag proteins from the cytoplasm to the nucleus. Internal deletion of the C-terminal portion of the Gag p10 region resulted in the nuclear sequestration of Gag and markedly diminished budding, suggesting that the nuclear export signal might reside within p10. Finally, we observed that a previously described matrix mutant, Myr1E, was insensitive to the effects of leptomycin B, apparently bypassing the nuclear compartment during virus assembly. Myr1E has a defect in genomic RNA packaging, implying that nuclear localization of Gag might be involved in viral RNA interactions. Taken together, these findings provide evidence that nuclear entry and egress of the Gag polyprotein are intrinsic components of the Rous sarcoma virus assembly pathway. PMID- 11891342 TI - How many named species are valid? AB - Estimates of biodiversity in both living and fossil groups depend on raw counts of currently recognized named species, but many of these names eventually will prove to be synonyms or otherwise invalid. This difficult bias can be resolved with a simple "flux ratio" equation that compares historical rates of invalidation and revalidation. Flux ratio analysis of a taxonomic data set of unrivalled completeness for 4,861 North American fossil mammal species shows that 24-31% of currently accepted names eventually will prove invalid, so diversity estimates are inflated by 32-44%. The estimate is conservative compared with one obtained by using an older, more basic method. Although the degree of inflation varies through both historical and evolutionary time, it has a minor impact on previously published background origination and extinction rates. Several lines of evidence suggest that the same bias probably affects more poorly studied, hyperdiverse living groups such as fungi and insects. If so, then current estimates of total global diversity could be revised downwards to as low as 3.5 10.5 million species. PMID- 11891343 TI - Reelin function in neural stem cell biology. AB - In the adult brain, neural stem cells (NSC) must migrate to express their neuroplastic potential. The addition of recombinant reelin to human NSC (HNSC) cultures facilitates neuronal retraction in the neurospheroid. Because we detected reelin, alpha3-integrin receptor subunits, and disabled-1 immunoreactivity in HNSC cultures, it is possible that integrin-mediated reelin signal transduction is operative in these cultures. To investigate whether reelin is important in the regulation of NSC migration, we injected HNSCs into the lateral ventricle of null reeler and wild-type mice. Four weeks after transplantation, we detected symmetrical migration and extensive neuronal and glial differentiation of transplanted HNSCs in wild-type, but not in reeler mice. In reeler mice, most of the injected HNSCs failed to migrate or to display the typical differentiation pattern. However, a subpopulation of transplanted HNSCs expressing reelin did show a pattern of chain migration in the reeler mouse cortex. We also analyzed the endogenous NSC population in the reeler mouse using bromodeoxyuridine injections. In reeler mice, the endogenous NSC population in the hippocampus and olfactory bulb was significantly reduced compared with wild type mice; in contrast, endogenous NSCs expressed in the subventricular zonewere preserved. Hence, it seems likely that the lack of endogenous reelin may have disrupted the migration of the NSCs that had proliferated in the SVZ. We suggest that a possible inhibition of NSC migration in psychiatric patients with a reelin deficit may be a potential problem in successful NSC transplantation in these patients. PMID- 11891344 TI - Conflicting levels of selection in the accumulation of mitochondrial defects in Saccharomyces cerevisiae. AB - The somatic accumulation of defective mitochondria causes human degenerative syndromes, senescence in fungi, and male sterility in plants. These diverse phenomena may result from conflicts between natural selection at different levels of organization. Such conflicts are fundamental to the evolution of cooperating groups, from cells to populations. We present a model in which defective mitochondrial genomes accumulate because of a within-cell replication advantage when among-cell selection for efficient respiration is relaxed. We tested the model by using experimental populations of the yeast Saccharomyces cerevisiae. We constructed yeast strains that were heteroplasmic for mitochondrial mutations that destroy the ability to respire (the petite phenotype) and followed the accumulation of mitochondrial defects in cultures with different effective population sizes. As predicted by the model, the inability to respire evolved only in small populations of S. cerevisiae, where among-cell selection favoring cells that can respire was reduced relative to within-cell selection favoring parasitic mitochondria. In a control experiment, mitochondrial point mutations that confer resistance to chloramphenicol showed no tendency to change in frequency under any culture conditions. The accumulation of some mitochondrial defects is therefore an evolutionary process, involving multiple levels of selection. The relative intensities of within- and among-cell selection may also explain the tissue specificity of human mitochondrial defects. PMID- 11891345 TI - An unexpected role for brain-type sodium channels in coupling of cell surface depolarization to contraction in the heart. AB - Voltage-gated sodium channels composed of pore-forming alpha and auxiliary beta subunits are responsible for the rising phase of the action potential in cardiac muscle, but the functional roles of distinct sodium channel subtypes have not been clearly defined. Immunocytochemical studies show that the principal cardiac pore-forming alpha subunit isoform Na(v)1.5 is preferentially localized in intercalated disks, whereas the brain alpha subunit isoforms Na(v)1.1, Na(v)1.3, and Na(v)1.6 are localized in the transverse tubules. Sodium currents due to the highly tetrodotoxin (TTX)-sensitive brain isoforms in the transverse tubules are small and are detectable only after activation with beta scorpion toxin. Nevertheless, they play an important role in coupling depolarization of the cell surface membrane to contraction, because low TTX concentrations reduce left ventricular function. Our results suggest that the principal cardiac isoform in the intercalated disks is primarily responsible for action potential conduction between cells and reveal an unexpected role for brain sodium channel isoforms in the transverse tubules in coupling electrical excitation to contraction in cardiac muscle. PMID- 11891347 TI - Molecular BKology: the study of splicing and dicing. AB - The activity of ion channels is regulated by several different mechanisms. Studies have been undertaken to determine how the large-conductance Ca(2+) sensitive K(+) channel (the BK channel) is regulated. Fury et al. discuss how the presence or absence of alternatively spliced regions in the BK channel alpha subunit can act as a molecular switch by which different kinases activate the BK channel. PMID- 11891346 TI - Voltage opens unopposed gap junction hemichannels formed by a connexin 32 mutant associated with X-linked Charcot-Marie-Tooth disease. AB - The X-linked form of Charcot-Marie-Tooth disease (CMTX) is an inherited peripheral neuropathy that arises in patients with mutations in the gene encoding the gap junction protein connexin 32 (Cx32), which is expressed by Schwann cells. We recently showed that Cx32 containing the CMTX-associated mutation, Ser-85-Cys (S85C), forms functional cell-cell channels in paired Xenopus oocytes. Here, we describe that this mutant connexin also shows increased opening of hemichannels in nonjunctional surface membrane. Open hemichannels may damage the cells through loss of ionic gradients and small metabolites and increased influx of Ca(2+), and provide a mechanism by which this and other mutant forms of Cx32 may damage cells in which they are expressed. Evidence for open hemichannels includes: (i) oocytes expressing the Cx32(S85C) mutant show greatly increased conductance at inside positive potentials, significantly larger than in oocytes expressing wild-type Cx32 (Cx32WT); and (ii) the induced currents are similar to those previously described for several other connexin hemichannels, and exhibit slowly developing increases with increasing levels of positivity and reversible reduction when intracellular pH is decreased or extracellular Ca(2+) concentration is increased. Although increased currents are seen, oocytes expressing Cx32(S85C) have lower levels of the protein in the surface and in total homogenates than do oocytes expressing Cx32WT; thus, under the conditions examined here, hemichannels in the surface membrane formed of the Cx32(S85C) mutant have a higher open probability than hemichannels formed of Cx32WT. This increase in functional hemichannels may damage Schwann cells and ultimately lead to loss of function in peripheral nerves of patients harboring this mutation. PMID- 11891348 TI - The banality of tobacco deaths. PMID- 11891349 TI - It is time to abandon youth access tobacco programmes. PMID- 11891351 TI - Planning to become a mom? PMID- 11891362 TI - Responses to tobacco control policies among youth. AB - OBJECTIVE: Explore adolescents' response to current and potential tobacco control policy issues. DESIGN: The 13 site Tobacco Control Network (TCN), sponsored by the Centers for Disease Control and Prevention, conducted 129 sex and ethnic homogeneous focus groups. PARTICIPANTS: 785 white, African American, Asian American/Pacific Islander, American Indian, and Hispanic adolescents who were primarily smokers from rural, urban, and suburban locations across the USA. MAIN OUTCOME MEASURES: Awareness, knowledge, opinions, and behaviour regarding laws and rules, prices, cigarette ingredients, and warning labels. RESULTS: Teenagers were generally familiar with laws and rules about access and possession for minors, but believed them ineffective. They were knowledgeable about prices, and reported that a sharp and sudden increase could lead them to adjust their smoking patterns but could also have negative consequences. They found a list of chemical names of cigarette ingredients largely meaningless, but believed that disclosing and publicising their common uses could be an effective deterrent, especially for those who were not yet smoking. They were aware of current warning labels, but considered them uninformative and irrelevant. CONCLUSIONS: Understanding teenagers' attitudes and behaviours before implementing policies that will affect them will likely increase their effectiveness. Disclosing and publicising the chemical contents of cigarettes, and increasing prices quickly and sharply, are potentially effective areas for policy change to impact adolescent tobacco use. PMID- 11891364 TI - Cigarette acquisition and proof of age among US high school students who smoke. AB - OBJECTIVE: To determine how US high school students who are under 18 years of age and who smoke obtain their cigarettes and whether they are asked for proof of age. DESIGN AND SETTING: Data from the Centers for Disease Control and Prevention's 1995, 1997, and 1999 national Youth Risk Behavior Surveys which employed national probability samples of students in grades 9-12 (ages 14-18 years). MAIN OUTCOME MEASURES: Associations of usual source of cigarettes and request for proof of age with variables such as sex, race/ethnicity, grade, and frequency of smoking. RESULTS: In 1999, among current smokers under age 18 years, 23.5% (95% confidence interval (CI), -4.5% to +4.5%) usually purchased their cigarettes in a store; among these students, 69.6% (95% CI -5.7% to +5.7%) were not asked to show proof of age. As days of past month smoking increased, reliance on buying cigarettes in a store (p < 0.001) and giving someone else money to buy cigarettes (p < 0.001) increased, and usually borrowing cigarettes decreased (p < 0.001). From 1995 to 1999, relying on store purchases significantly decreased (from 38.7% (95% CI -4.6% to + 4.6%) to 23.5% (95% CI -4.5% to +4.5%)); usually giving someone else money to buy cigarettes significantly increased (from 15.8% (95% CI -3.6% to +3.6%) to 29.9% (95% CI -4.5% to + 4.5%)). CONCLUSIONS: Stricter enforcement of tobacco access laws is needed to support other community and school efforts to reduce tobacco use among youth. Furthermore, effective interventions to reduce non-commercial sources of tobacco, including social, need to be developed and implemented. PMID- 11891365 TI - Application of a rating system to state clean indoor air laws (USA). AB - OBJECTIVE: To develop and implement a system for rating state clean indoor air laws. DESIGN: The public health interest of state clean indoor air laws is to limit non-smoker exposure to environmental tobacco smoke (ETS). Current estimates of health risks and methods available for controlling ETS provided a framework for devising a ratings scale. An advisory committee applied this scale to each of seven site specific smoking restrictions and two enforcement related items. For each item, a target score of +4 was identified. The nine items were then combined to produce a summary score for each state. A state that achieved the target across all nine items would receive a summary score of 36 points and be eligible to receive an additional 6 points for exceeding the target on six of the nine items, resulting in a maximum summary score of 42 points. Individual scores were also adjusted to reflect state level preemption measures. Each state's law was evaluated annually from 1993 through 1999. SETTING: USA. MAIN OUTCOME MEASURE: A summary score measuring the extensiveness of the state's clean indoor air law. RESULTS: State laws restricting smoking in the seven individual locations of interest were relatively weak. The overall mean score across the location restrictions ranged from 0.72 in 1993 to 0.98 in 1999. Mean scores were higher for the enforcement items than for the location restrictions. Summary scores ranged from 0 to 20 in 1993 and 0 to 31 in 1994 through 1999. Average summary scores ranged from 8.71 in 1993 to 10.98 in 1999. By the end of 1999, scores increased for 22 states; however, between 1995 and 1997 there were no changes in the summary scores. Three states scored zero points across all years. From 1993 through 1999, there was a 41% increase in the number of states that had in place state level preemption measures. CONCLUSION: The number of newly enacted state clean indoor air laws has remained relatively stagnant since 1995. With a few exceptions, as of the end of 1999, progress in enacting state laws to meet specified public health targets for reducing exposure to ETS was relatively low. Thus, state laws in the USA provide, on average, only minimal protection in specified areas and, given the increase in preemption, are increasingly undermining those passed in localities. PMID- 11891366 TI - Trends and affordability of cigarette prices: ample room for tax increases and related health gains. AB - BACKGROUND: Increasing the price of tobacco products is arguably the most effective method of curbing the prevalence and consumption of tobacco products. Price increases would reduce the global burden of disease brought about by tobacco consumption. OBJECTIVES: To compare cigarette price data from more than 80 countries using varying methods, examine trends in prices and affordability during the 1990s, and explore various policy implications pertaining to tobacco prices. DESIGN: March 2001 cigarette price data from the Economist Intelligence Unit are used to compare cigarette prices across countries. To facilitate comparison and to assess affordability, prices are presented in US dollars, purchasing power parity (PPP) units using the Big Mac index as an indicator of PPP and in terms of minutes of labour required to purchase a pack of cigarettes. Annual real percentage changes in cigarette prices between 1990 and 2000 and annual changes in the minutes of labour required to buy cigarettes between 1991 and 2000 are also calculated to examine trends. RESULTS: Cigarette prices tend to be higher in wealthier countries and in countries that have strong tobacco control programmes. On the other hand, minutes of labour required to purchase cigarettes vary vastly between countries. Trends between 1990 and 2000 in real prices and minutes of labour indicate, with some exceptions, that cigarettes have become more expensive in most developed countries but more affordable in many developing countries. However, in the UK, despite recent increases in price, cigarettes are still more affordable than they were in the 1960s. CONCLUSIONS: The results suggest that there is ample room to increase tobacco prices through taxation. In too many countries, cigarette prices have failed to keep up with increases in the general price level of goods and services, rendering them more affordable in 2000 than they were at the beginning of the decade. Opportunities to increase government revenue and improve health through reduced consumption brought about by higher prices have been overlooked in many countries. PMID- 11891367 TI - WHO Europe evidence based recommendations on the treatment of tobacco dependence. PMID- 11891368 TI - Examining the effects of tobacco treatment policies on smoking rates and smoking related deaths using the SimSmoke computer simulation model. AB - OBJECTIVES: To develop a simulation model to predict the effects of different smoking treatment policies on quit rates, smoking rates, and smoking attributable deaths. METHODS: We first develop a decision theoretic model of quitting behaviour, which incorporates the decision to quit and the choice of treatment. A model of policies to cover the costs of different combinations of treatments and to require health care provider intervention is then incorporated into the quit model. The policy model allows for the smoker to substitute between treatments and for policies to reduce treatment effectiveness. The SimSmoke computer simulation model is then used to examine policy effects on smoking rates and smoking attributable deaths. RESULTS: The model of quit behaviour predicts a population quit rate of 4.3% in 1993, which subsequently falls and then increases in recent years to 4.5%. The policy model suggests a 25% increase in quit rates from a policy that mandates brief interventions and the coverage of all proven treatments. Smaller effects are predicted from policies that mandate more restricted coverage of treatments, especially those limited to behavioural treatment. These policies translate into small reductions in the smoking rate at first, but increase to as much as a 5% reduction in smoking rates. They also lead to substantial savings in lives. CONCLUSIONS: Tobacco treatment policies, especially those with broad and flexible coverage, have the potential to increase smoking cessation substantially and decrease smoking rates in the short term, with fairly immediate reductions in deaths. PMID- 11891369 TI - Cigarette nicotine yields and nicotine intake among Japanese male workers. AB - OBJECTIVES: To analyse brand nicotine yield including "ultra low" brands (that is, cigarettes yielding less-than-or-equal 0.1 mg of nicotine by Federal Trade Commission (FTC) methods) in relation to nicotine intake (urinary nicotine, cotinine and trans-3'-hydroxycotinine) among 246 Japanese male smokers. DESIGN: Cross sectional study. SETTING: Two companies in Osaka, Japan. SUBJECTS: 130 Japanese male workers selected randomly during their annual regular health check up and 116 Japanese male volunteers taking part in a smoking cessation programme. MAIN OUTCOME MEASUREMENTS: Subjects answered a questionnaire about smoking habits. Following the interview, each participant was asked to smoke his own cigarette and, after extinguishing it, to blow expired air into an apparatus for measuring carbon monoxide concentration. Urine was also collected for the assays of nicotine metabolites. RESULTS: We found wide variation in urinary nicotine metabolite concentrations at any given nicotine yield. Based on one way analysis of variance (ANOVA), the urinary nicotine metabolite concentrations of ultra low yield cigarette smokers were significantly lower compared to smokers of high (p = 0.002) and medium yield cigarettes (p = 0.017). On the other hand, the estimated nicotine intake per ultra low yield cigarette smoked (0.59 mg) was much higher than the 0.1 mg indicated by machine. CONCLUSIONS: In this study of Japanese male smokers, actual levels of nicotine intake bore little relation to advertised nicotine yield levels. Our study reinforces the need to warn consumers of inappropriate advertisements of nicotine yields, especially low yield brands. PMID- 11891370 TI - Long term and transitional intermittent smokers: a longitudinal study. AB - OBJECTIVE: To investigate differences in snuff consumption, sociodemographic and psychosocial characteristics between baseline intermittent smokers that had become daily smokers, stopped smoking or remained intermittent smokers at the one year follow up. DESIGN/SETTING/PARTICIPANTS/MEASUREMENTS: A population of 12 507 individuals interviewed at baseline in 1992-94 and at a one year follow up, aged 45-69 years, was investigated in a longitudinal study. The three groups of baseline intermittent smokers were compared to the reference population (all others) according to sociodemographic, psychosocial, and snuff consumption characteristics. A multivariate logistic regression model was used to assess differences in psychosocial conditions, adjusting for age, sex, country of origin, marital status, education, and snuff consumption. RESULTS: 60% of all baseline intermittent smokers had remained intermittent smokers, 16% had become daily smokers, and 24% had stopped smoking at the one year follow up. The long term intermittent smokers and those who had stopped smoking were young, unmarried, highly educated, and snuff consumers to a higher extent than the reference population. They also had more psychosocial resources than the reference population, while the psychosocial resources of those who had become daily smokers were poorer. CONCLUSIONS: The majority of intermittent smokers are long term intermittent smokers. The results suggest that long term intermittent smokers have other psychosocial characteristics than daily smokers. PMID- 11891371 TI - Tobacco industry documents: comparing the Minnesota Depository and internet access. AB - OBJECTIVE: To assess the comparability of searches conducted on two publicly available tobacco industry document collections: hard copies housed and maintained by a neutral party in the Minnesota Depository and electronic copies available through tobacco industry maintained websites. METHODS: We conducted a set of searches in Minnesota and then conducted the same searches using the industry websites. We matched documents by Bates number, weeded out duplicates, and coded documents that were unique to either collection as major, minor, or trivial. RESULTS: Among hundreds of documents produced by several searches, we found only four unique major documents in the Minnesota Depository. By contrast, we found 62 unique major documents using the websites. CONCLUSION: These results suggest that researchers can rely on industry websites while waiting for improved access resulting from searching, indexing, and document storage administered by the tobacco control community. Searching the tobacco industry websites is at least as good as searching in Minnesota and may in some instances actually be better. Four smaller subcollections, however, can only be searched by hand in Minnesota. PMID- 11891372 TI - Controversies in tobacco control: the limitations of fear messages. PMID- 11891373 TI - The continuing importance of emotion in tobacco control media campaigns: a response to Hastings and MacFadyen. PMID- 11891375 TI - Hungary introduces a total ban on tobacco advertising. PMID- 11891376 TI - Tobacco use among school personnel in Bihar, India. PMID- 11891377 TI - Exposure to environmental tobacco smoke in public places in Barcelona, Spain. PMID- 11891378 TI - Ophthalmologists' and optometrists' attitudes and behaviours regarding tobacco cessation intervention. PMID- 11891380 TI - Monitoring of influenza in the EISS European network member countries from October 2000 to April 2001. AB - In countries covered by the European Influenza Surveillance Scheme (EISS), the 2000-2001 winter was marked mainly by the spread of influenza A(H1N1) viruses. Influenza B, which globally represented a minority of cases, was common later in the season and predominant in Great Britain, Ireland, and Portugal. Influenza activity was at its maximum during the period of January and February/March 2001 with little time lag between countries (maximum four weeks). Overall, the morbidity rates reported were much lower than for the previous season, illustrating a moderate level of influenza activity. PMID- 11891379 TI - A smoking cessation telephone resource: feasibility and preliminary evidence on the effect on health care provider adherence to smoking cessation guidelines. PMID- 11891381 TI - Influenza pandemic planning in Europe. AB - The World Health Organization strongly recommends that all countries prepare in advance multidisciplinary pandemic plans to prevent and control the next influenza pandemic. We carried out a survey of influenza surveillance methods among members of the European Influenza Surveillance Schemes, EISS, which included a set of questions on pandemic planning. All but one of the countries have a pandemic plan or are in the process of producing one. A coordination of these different national plans at a European level would probably contribute to their improved impact and efficiency. PMID- 11891382 TI - Malaria incidence and mortality in Italy in 1999-2000. AB - In 1999-2000, a total of 2060 malaria cases were reported by the ISS. Most of the patients took inappropriate treatments or did not have any prophylaxis. Ninety three per cent became infected in African malarious countries, 4% in Asian countries, and 3% in Latin America. P. falciparum accounted for 84% of the cases, followed by P. vivax (8%), P. ovale (5%), and P. malariae (2%). Deaths corresponded to an annual case fatality rate of 0.3% in 1999 and 0.5% in 2000. In general, imported malaria cases reflect the number of Italian travellers who underestimate the infection risk in Asian and Latin American malarious countries and permanent residents of African origin who visit their relatives in their native countries. PMID- 11891383 TI - Escherichia coli O157 infections and unpasteurised milk. AB - We report on two children with Escherichia coli O157 infection, one of whom developed haemolytic uraemic syndrome (HUS). Both had drunk raw cows or goats milk in the week before their illness. Molecular subtyping identified a sorbitol fermenting Escherichia coli O157:H isolate from a dairy cow. This isolate differed from Shiga toxin producing O157:H strains isolated from the 6 year old boy with HUS. This result underlines the need to search for other causes of infection, despite documented consumption of unpasteurised milk. In the second patient, human sorbitol non-fermenting O157:H isolates and animal isolates from goats were indistinguishable. The isolation of indistinguishable sorbitol non fermenting Escherichia coli O157:H from contact animals supports the association between HUS and consumption of raw goats milk, and re-emphasises the importance of pasteurising milk. PMID- 11891384 TI - Imported rocket salad partly responsible for increased incidence of hepatitis A cases in Sweden, 2000-2001. AB - An increased incidence of domestic hepatitis A without any obvious source of infection in Sweden and a small outbreak in late spring 2001 led to the undertaking of a case-control study. Consumption of imported rocket salad was clearly associated with disease (odds ratio 9.1, 95% confidence interval 1.5 to 69). The importation of vegetables from countries where hepatitis A is endemic to countries where this disease is not endemic continues to be a public health problem. PMID- 11891385 TI - Bioterrorism: crime and opportunity. AB - The unprecedented and tragic terrorist attacks in the United States (US) have sent shock waves through national administrations that have grown accustomed to fighting expenditure wars in the health area and had relegated public health vigilance and emergency preparedness to the back burner. It was obvious from the immediate reaction to the horrors and menaces of the autumn of 2001 that, insofar as health and safety is concerned, governments continued to measure success by the degree of quietness, remoteness and uneventful normality that is achieved by those entrusted with the responsibility to protect health. The paradox of health and safety is that you are winning when you hear nothing: any publicity is bound to be bad publicity. PMID- 11891386 TI - Bioterrorism preparedness and response in European public health institutes. AB - The terrorist attacks on 11 September 2001 and the deliberate release of anthrax in the United States had consequences for public health not only there, but also in Europe. Europe's public health systems had to manage numerous postal materials possibly contaminated with anthrax. Our survey aimed to document the response of European public health institutes to recent bioterrorist events to identify the gaps that need to be addressed; 18 institutes from 16 countries participated in this Euroroundup. Bioterrorist threats in Europe were hoaxes only, and should be considered as a "preparedness exercise" from which three lessons can be drawn. Firstly, because of inadequate preparedness planning and funding arrangements, Europe was not ready in October 2001 to respond to bioterrorism. Secondly, although European institutes reacted quickly and adapted their priorities to a new type of threat, they need adequate and sustained support from national governments to maintain their overall capacity. Thirdly, the recent crisis demonstrated the need for increased investment in epidemiology training programmes and the establishment of a technical coordination unit for international surveillance and outbreak response in the European Union. PMID- 11891387 TI - Deliberate releases of biological agents: initial lessons for Europe from events in the United States. AB - The experience of autumn 2001, when anthrax spores were released in the postal system, had considerable consequences in the United States and in Europe. The threat of covert deliberate releases against civilians has become a reality. In Europe, despite the growing number of criminal hoaxes, no cases of anthrax linked to deliberate releases have been reported, and the only contaminated letters were addressed to American embassies abroad. Nevertheless, the time has come for Europe to set up efficient and timely plans to respond to bioterrorism, under the coordination of the European Commission. PMID- 11891388 TI - The use of smallpox virus as a biological weapon: the vaccination situation in France. AB - In the context of its plan to fight against bioterrorism, the Ministry of Health asked the Institut de Veille Sanitaire to evaluate the epidemic risk from a release of the small pox virus, and to make recommendations on potential vaccination strategies to be implemented. A benefit/risk assessment of various vaccination scenarios, including vaccination of the whole French population, was carried out to evaluate the severity of a terrorist action threat. This analysis concludes that at this stage, vaccination action does not seem to be justified. Even in the case of a real threat, the vaccination of frontline healthcare personnel, and in particular of contacts of cases, must be given priority. PMID- 11891389 TI - Isolation of Guinea pig inner hair cells using manual microsurgical dissection. AB - Cochlear inner hair cells (IHCs) and outer hair cells (OHCs) show distinctive morphological features that are usually sufficient to distinguish these two species in vitro. However, OHCs may sometimes resemble IHCs when they are mechanically distorted or begin swelling at their basal end. As a result, accurately discriminating the cells based on morphology was thought to be problematic. An objective method that allows us to clearly and unambiguously distinguish these two cell types is therefore of continued interest. We describe a protocol in which solitary IHCs were harvested from guinea pig cochlea using a manual microsurgical dissection. PMID- 11891390 TI - Short-term outcome and prognosis of acute low-tone sensorineural hearing loss by administration of steroid. AB - Acute low-tone hearing loss (ALHL) is a typical type of hearing loss in Meniere's disease and thought to be caused by endolymphatic hydrops in the inner ear. We treated 40 patients with ALHL by administration of the steroid and the early outcome and prognosis of the hearing level was retrospectively evaluated. The prognosis was generally determined within 7-10 days after administration of steroid. High-dose steroid cured some patients who failed to recover with low dose steroid therapy. Our results showed that steroid is one of the effective therapies for ALHL and supported that etiology of ALHL involves an immune response. PMID- 11891391 TI - Endoillumination-guided intranasal microscopic dacryocystorhinostomy for difficult cases. AB - PURPOSE: This prospective study explores the efficacy of a combined approach by intranasal microsurgical dacryocystorhinostomy and fiberglass endoillumination of the tear passage for difficult cases of lacrimal sac or nasolacrimal duct obstruction. METHODS: Microscopic dacryocystorhinostomy was performed in 67 patients with lacrimal obstruction and for revision of external dacryocystorhinostomy. Improved endonasal visualization by microscope and combination with endoillumination of the tear passage allows instant identification and opening of the lacrimal nasal sac without skin incision. Silicon tubes were used for bicanalicular intubation. The patients were prospectively observed for at least 1 year. RESULTS: The operation was well tolerated by all patients. In 95%, a persistently open tear passage was successfully established. No serious complications were observed. CONCLUSION: Endoillumination of the lacrimal sac is useful for locating the sac precisely and refining the endonasal operation technique. PMID- 11891392 TI - Evaluation of the T&T olfactometer by mapping c-fos protein in an olfactory bulb. AB - OBJECTIVES: Although in Japan the T&T olfactometer is most commonly used in patients suffering from smelling disorders, no scientific analysis has been performed so far. The objective of this study was to clarify whether the five odorants used in the T&T olfactometer are suitable or not. We mapped the glomeruli activated by the five test odorants employed in the T&T olfactometer. The expression of c-fos protein as a marker of neuronal excitation was monitored using an immunohistochemical technique. STUDY DESIGN: The expression of c-fos protein in the rat olfactory bulb was investigated to determine what part of the olfactory system is activated by five odorants used in the T&T olfactometer. METHODS: Each rat was isolated in a clean cage for 120 min to reduce the basal expression of c-fos protein. Each rat was fixed in the cage and exposed to one of five test odorants for 90 min. The expression of c-fos protein was measured using an immunohistochemical technique. RESULTS: Each odorant activated numerous glomeruli and the patterns of distribution of activated glomeruli were specific to each odorant. Glomeruli in most regions of the bulb were activated by all five test odorants. CONCLUSION: We assume that in the T&T olfactometer the optimal ((five)) odorants are employed, because the glomeruli activated by those odorants showed unique patterns in the immunohistochemical assay. PMID- 11891393 TI - Improving the reproducibility of acoustic rhinometry in the assessment of nasal function. AB - Acoustic rhinometry readings are very position dependent, and it was hypothesized that this accounts for its relative lack of reproducibility on a day-to-day basis. Multiple readings on each visit were taken to investigate their impact, if any, on improving the day-to-day reproducibility of the method. Measurements of the minimal cross-sectional area of the nose as measured by acoustic rhinometry were studied in 10 subjects following nasal decongestion. For each individual, acoustic rhinometry was performed ten times. The ten recordings were repeated again, in an identical manner on a separate day. The subjects were repositioned and the nasal probes reinserted between each measurement. The mean coefficient of variation for minimal cross-sectional area readings in all 10 subjects was calculated as 9.92%. This is comparable to the day-to-day variability of acoustic rhinometry as measured by other workers and thus supports the hypothesis that the high measurement error of the device (rhinometer) is a function of positional variation during data acquisition. We were able to demonstrate a minimal gain in intervisit reproducibility by doing multiple recordings per person, with a plateau effect of reproducibility after 7 repeat readings. PMID- 11891394 TI - Acute exudative inflammation and nasally exhaled nitric oxide are two independent phenomena. AB - BACKGROUND: Increased levels of nitric oxide (NO) and the exudations of plasma proteins to the airway lumen have both been considered characteristics of airway inflammation. The aim of the present study was to investigate a possible relationship between nasal NO concentrations and acutely induced exudative inflammation of the nasal mucosa. METHODS: Twelve healthy non-allergic subjects participated. Nasal challenges with saline, histamine 40 microg/ml (H1), histamine 400 microg/ml (H2), oxymethazoline, 0.25 mg/ml (OXY), and a combination of oxymethazoline 0.25 mg/ml and histamine 800 microg/ml (OXYH), were performed on separate occasions. Exhaled NO was measured after each challenge, and alpha(2) macroglobulin (as a marker of plasma exudation) was measured in nasal lavage fluids after the H1 and H2 challenges. RESULTS: The mean baseline NO in all measurements was 164 plus minus 10.3 ppb. Saline and H1 challenge did not change NO and alpha(2)-macroglobulin levels. H2 challenge showed a tendency to reduce NO levels, and the most pronounced decrease was seen after 10 min (-36.3 +/- 16.3%, p = 0.07). This reduction was sustained throughout the registration period. Simultaneously with the decrease in NO, alpha(2)-macroglobulin levels were increased significantly. OXY challenge alone reduced NO significantly throughout the whole registration period. Maximum decrease was seen at 40 min (-21.3 plus minus 3.4%, p = 0.03). The OXYH challenge also reduced NO, with a maximal reduction recorded at 10 min (-29.4 +/- 6.4%, p = 0.03). The reduction of NO was sustained throughout the registration period (p < 0.01). CONCLUSION: Histamine 400 microg/ml induced a prompt plasma exudation response whereas a decrease in nasal NO was registered, suggesting that these two events are not necessarily linked. Furthermore it was shown that the vasoconstrictor oxymethazoline reduced nasal NO, which could be related to reduced mucosal blood flow, whereas the reduction of nasal NO after histamine challenge remains to be elucidated. PMID- 11891395 TI - Percutaneous endoscopic gastrostomy in head and neck cancer patients. AB - OBJECTIVE: To present our experience with the indications and complications of percutaneous endoscopic gastrostomy (PEG) in head and neck cancer patients. MATERIALS AND METHODS: In a retrospective study of the patients treated, we reviewed the records of 43 patients diagnosed with head and neck cancer at the Otorhinolaryngology Department, in which a PEG was performed by the Unit of Digestive Endoscopy. RESULTS: All cases had squamous cell carcinoma. Larynx was the most frequent primary site, with 21 cases (49%), followed by hypopharynx, 12 patients (28%). Indications for PEG were: dysphagia (53.5%) and pharyngocutaneous fistula (43.5%). The most frequent complication was a local infection. CONCLUSION: PEG is a good choice for long-term enteral feeding in head and neck cancer patients due to its low complication rate and easy handling. PMID- 11891396 TI - Cryptococcal meningitis: implications for the otologist. AB - Cryptococcal meningitis can present to the otologist with hearing loss and vestibular dysfunction. A man with cryptococcal meningitis presenting with bilateral profound sensorineural hearing loss and vestibular dysfunction is described. The difficulty in arriving at the diagnosis and consequences of misdiagnosis are discussed. Histological and clinical studies suggest that the cochlear nerve is likely to be damaged in patients who have deafness associated with this disease. Retrocochlear damage may result in cochlear implantation having a poor outcome. PMID- 11891397 TI - Bone wax foreign body granuloma in the mastoid. AB - The sue of bone wax is commonly used to control bleeding during mastoid surgery. An unusual case of bone wax granuloma in the mastoid, with sigmoid sinus thrombosis, is reported. Although the use of bone wax in and around the mastoid is generally considered safe with few complications, caution should be exercised, particularly in infected fields and in patients known to have general immunohypersensitivity. PMID- 11891398 TI - Psychogenic hearing loss with panic anxiety attack after the onset of acute inner ear disorder. AB - A very rare case of a 50-year-old female showing psychogenic hearing loss with a panic anxiety attack that complicated an acute organic sensorineural hearing loss is reported. At the first visit to our clinic, the patient showed left sensorineural hearing loss with an inner ear disorder pattern. Five days after the onset, her left hearing threshold markedly increased without any subjective signs. On the next day, she suddenly experienced a severe panic attack with anxiety. After the attack, she felt mildly anxious and depressed. A combined therapy using primary corticosteroid therapy for the acute inner ear disorder, psychiatric counseling based on cognitive therapy and the administration of a minor tranquilizer was performed. Her left hearing threshold recovered to within normal ranges except in the high-frequency ranges immediately after the treatment. This case was considered very rare because: (1) the panic anxiety attack occurred in the conversion disorder as psychogenic hearing loss and (2) the psychogenic hearing loss complicated the primary sudden deafness. We suggest that otorhinolaryngologists should have psychiatric knowledge and be able to treat psychogenic hearing loss as a primary care. PMID- 11891399 TI - Delayed facial nerve palsy following uneventful stapedectomy. AB - Facial nerve palsy is a rare complication of stapedectomy. Its onset may be immediate or delayed by several days. The authors present a case of a 59-year-old man who developed right delayed peripheral facial nerve palsy occurring after uncomplicated ipsilateral stapedectomy. The incidence, treatment and prognosis of this complication are discussed. PMID- 11891400 TI - Horner's syndrome and thyroid neoplasms. AB - Although thyroid goiter is a common condition, it rarely results in Horner's syndrome. We report a case of a patient with an intrathoracic multinodular goiter complicated by Horner's syndrome. Benign thyroid disease was confirmed pathologically, and the patient's symptoms improved after surgery. In the literature, the major cause of Horner's syndrome is neoplasia, with malignant lesions being twice as frequent as benign tumors. An extensive review of the literature demonstrates a different repartition for thyroid neoplasia: including our case, 38 cases of Horner's syndrome secondary to a benign thyroid tumor are described, against only 8 cases caused by a thyroid carcinoma. We conclude that contrary to the commonly held opinion, Horner's syndrome is more often due to benign thyroid diseases than to thyroid malignancies. PMID- 11891401 TI - Functional upper airway obstruction in a child with Freeman-Sheldon syndrome. AB - Freeman-Sheldon syndrome is defined as a combination of microstomia, deep set eyes, small palpebral fissures, arthrogryposis with ulnar deviation of the hand, talipes equinovarus and generalized muscular hypertension. Respiratory and swallowing problems are frequently encountered in these patients due to small orifices of mouth and nose. Obstruction of the upper airway tract resulting in tracheostomy has only been described twice. The described child manifested the typical dysmorphic features of Freeman-Sheldon syndrome and suffered from serious respiratory distress and swallowing difficulties from birth. The boy died at the age of 7 months after accidental decannulation of the tracheostoma during sleep. He did not show anatomical or histopathological abnormalities in the pharyngeal, laryngeal or tracheal regions. We assume that the only explanation of the repeated obstructive episodes is a functional muscular obstruction. PMID- 11891403 TI - The IKK/NF-kappaB pathway. PMID- 11891402 TI - Inflammatory myositis. PMID- 11891404 TI - Macrophage migration inhibitory factor. AB - Macrophage migration inhibitory factor (MIF) has been proposed to be the physiologic counter-regulator of glucocorticoid action within the immune system. In this role, MIF's position within the cytokine cascade is to act in concert with glucocorticoids to control both the "set point" and the magnitude of the inflammatory response. As well as overriding the immunosuppressive effects of glucocorticoids, it is now well established that MIF has a direct proinflammatory role in inflammatory diseases, such as sepsis, rheumatoid arthritis, and glomerulonephritis. The functions of MIF within the immune system are both unique and diverse, and although a unified molecular mechanism of action remains to be elucidated, there have been significant advances in our understanding of how MIF affects cellular processes. This review discusses the pathogenic role of MIF in inflammatory disease and highlights the novel structural, functional, and mechanistic properties of MIF. PMID- 11891405 TI - High-mobility group-I/Y proteins: Potential role in the pathophysiology of critical illnesses. AB - High-mobility group (HMG) proteins are architectural factors that have been shown to play a role in the transcriptional regulation of various mammalian genes. One family of HMG proteins, HMG-I/Y, is known to facilitate the initiation of gene transcription by modifying the conformation of DNA and recruiting transcription factors into an organized complex on transcriptional regulatory regions of specific genes. In many circumstances, the nuclear factor-kappaB family of transcription factors is involved in gene regulation that is mediated by HMG-I/Y. We will review the mechanisms by which HMG-I/Y proteins regulate gene transcription, give an overview of selected genes regulated by HMG-I/Y, summarize the potential roles of these genes in critical illnesses, and provide more detailed information about the role of HMG-I/Y in the regulation of nitric oxide synthase-2 during an inflammatory response, such as endotoxemia/sepsis. PMID- 11891406 TI - Endotoxin tolerance: A review. AB - Endotoxin tolerance was initially described when it was observed that animals survived a lethal dose of bacterial endotoxin if they had been previously treated with a sublethal injection. In animal models, two phases of endotoxin tolerance are described, an early phase associated with altered cellular activation and a late phase associated with the development of specific antibodies against the polysaccharide side chain of Gram-negative organisms. Recently, there has been a tremendous resurgence of interest in the mechanisms responsible for altered responsiveness to bacterial endotoxin. Host immune cells, particularly macrophages and monocytes, that are exposed to endotoxin for 3 to 24 hrs are rendered "tolerant" and manifest a profoundly altered response when rechallenged with bacterial endotoxin or lipopolysaccharide. The "lipopolysaccharide-tolerant" phenotype is characterized by inhibition of lipopolysaccharide-stimulated tumor necrosis factor production, altered interleukin-1 and interleukin-6 release, enhanced cyclooxygenase-2 activation, inhibition of mitogen-activated protein kinase activation, and impaired nuclear factor-kappaB translocation. Human monocytes and macrophages can be induced to become tolerant, and there is increasing evidence that monocytic cells from patients with systemic inflammatory response syndrome and sepsis have many characteristics of endotoxin tolerance. PMID- 11891407 TI - Mitogen-activated protein kinases. AB - The cellular control switches are regulated through an extensive network of interactive intracellular signal transduction pathways, such as the mitogen activated protein kinase (MAPK) family. The MAPK pathways may play an important role in the inappropriate inflammatory responses that lead to systemic inflammatory response syndrome or multiple organ dysfunction syndrome. Therefore, elucidating the activation status of the MAPK pathways may be a method to identify patients at risk for systemic inflammatory response syndrome/multiple organ dysfunction syndrome. Also, manipulating the proper pathways may improve patients' outcomes. However, the MAPK family is part of a complex interactive network, which may initiate an unpredictable reaction to the indiscriminate inhibition or activation of a single component. A major challenge is to elucidate the principles by which the network is assembled, so a more tissue- and temporal specific approach can be used. PMID- 11891408 TI - Phosphatases: Counterregulatory role in inflammatory cell signaling. AB - Cellular responses to external stimuli proceed through multiply complex signal transduction networks. The manner by which signals are propagated from the cell's surface to the nuclei is, in large part, dependent on the phosphorylation of signaling proteins mediated by kinases. As in most biological systems, this process of kinase-mediated phosphorylation is balanced by the presence of a dephosphorylating system comprised of a number of families of phosphatases. The purpose of this review is to describe the various members of the main classes of phosphatases and to examine their role in regulating signal transduction pathways relevant to critical illness. Because recent work has provided key insight into the role of kinase-mediated signaling pathways, such as the mitogen-activated protein kinase and inhibitor of nuclear factor-kappaB kinase pathways, in inflammatory states, emphasis has been placed on the regulation of these events. It is hoped that deriving novel insight into the regulatory phosphatases will allow a greater understanding of inflammatory cell signaling events and potentially identify novel sites for therapeutic intervention in the future. PMID- 11891409 TI - Interactions between the heat shock response and the nuclear factor-kappaB signaling pathway. AB - The heat shock response (HSR) and the nuclear factor (NF)-kappaB signaling pathway are two fundamental cellular responses. Various laboratories have documented in vitro and in vivo interactions between the HSR and NF-kappaB activation when they are activated sequentially. For example, induction of the HSR before a proinflammatory signal inhibits NF-kappaB activation and NF-kappaB dependent proinflammatory gene expression. The central point of control appears to be at the level of IkappaBalpha phosphorylation as demonstrated by HSR mediated inhibition of IkappaB kinase activation and HSR-mediated induction of intracellular phosphatase activity. In addition, induction of the HSR can independently increase de novo expression of the IkappaBalpha gene, thereby providing another potential mechanism through which the HSR can modulate cellular proinflammatory signaling. Another level of interaction is illustrated by the observation that various pharmacologic inhibitors of the NF-kappaB pathway are capable of simultaneously inducing the HSR. In direct contrast, induction of the HSR after a proinflammatory signal can lead to programmed cell death. Further understanding of how these two fundamental cellular responses interact at the molecular level holds the potential to elucidate some of the molecular interactions that occur during disease states common to critical care medicine. PMID- 11891412 TI - The APOE locus and the pharmacogenetics of lipid response. AB - Genetic variation at the APOE locus has been associated with plasma lipoprotein concentrations in the fasting (low-density, and high-density lipoproteins and triglycerides), and in the postprandial (triglyceride-rich lipoproteins) states. Resulting from these associations, the APOE locus has been found to be a significant genetic determinant of cardiovascular disease in the general population. Beyond the traditional association studies, APOE genetic variation has been shown to play a significant role, which explains some of the individual variations in therapies aimed at normalizing plasma lipid concentrations. Thus, the APOE E4 allele has been shown in some studies to be associated with increased response to dietary intervention. Conversely, APOE E2 carriers appear to be more responsive to statin therapy. The mechanisms behind these observations, however, have not been elucidated. Moreover, several other gene:environment and gene:therapy interactions have recently been demonstrated, thus further increasing the interest in this remarkable apolipoprotein. PMID- 11891413 TI - Molecular biology of apolipoprotein E. AB - Apolipoprotein E, first identified 26 years ago as a serum protein that mediates extracellular cholesterol transport, is now known to regulate multiple additional metabolic pathways. Several clinically important disorders of the vasculature and brain are differentially caused, or modified, by the three isoforms of this protein. Apolipoprotein E was previously believed to traffic exclusively through binding cell surface receptors, endocytosis, and hydrolysis. However, recent studies reveal a variety of additional physiologically important roles for apolipoprotein E that are mediated through interactions with different families of receptors, through binding other proteins, and through other intracellular trafficking pathways and second messengers. Much research is now directed toward identifying those pathways of apolipoprotein E metabolism that are differentially regulated by the various isoforms of apolipoprotein E, with the goal of identifying the particular molecular pathways that result in vascular and neurologic disorders. PMID- 11891414 TI - Apolipoprotein A-IV polymorphisms and diet-gene interactions. AB - Apolipoprotein A-IV is a 46kDa glycoprotein that is synthesized by intestinal enterocytes and is incorporated into the surface of nascent chylomicrons. Considerable evidence suggests that apolipoprotein A-IV plays a role in intestinal lipid absorption and chylomicron assembly. We have proposed that polymorphisms that alter the interfacial behavior of apolipoprotein A-IV may modulate the physical properties and metabolic fate of plasma chylomicrons. Of the reported genetic polymorphisms of apolipoprotein A-IV, two, Q360H and T347S, are known to occur at high frequencies among the world populations. Biophysical studies have established that the Q360H isoprotein displays higher lipid affinity; conversely the T347S isoprotein is predicted to be less lipid avid. Recent studies have shown that the Q360H polymorphism is associated with increased postprandial hypertriglyceridemia, a reduced low-density lipoprotein response to dietary cholesterol in the setting of a moderate fat intake, an increased high-density lipoprotein response to changes in total dietary fat content, and lower body mass and adiposity; the T347S polymorphism appears to confer the opposite effects. Studies on the diet-gene interactions of other apolipoprotein A-IV alleles are needed, as are studies on the interactions between apolipoprotein A-IV alleles and other apolipoprotein polymorphisms. PMID- 11891415 TI - Phospholipid transfer protein. AB - A role for phospholipid transfer protein (PLTP) in HDL remodelling and in the formation of pre-beta-HDL is now well established, both in vivo and in vitro. Over-expression of human PLTP in C57BL6 mice lowers plasma HDL levels, probably because of increased HDL catabolism. Despite these low HDL levels, plasma from these mice mitigates cholesterol accumulation in macrophages and has increased potential for pre-beta-HDL formation. Plasma HDL concentration is also decreased in PLTP knockout mice. These intriguing observations can be explained by recent studies that indicate that PLTP is not only involved in remodelling of HDL subfractions but also in VLDL turnover. The role of PLTP in atherogenesis and VLDL synthesis was demonstrated in transgenic mouse models with increased susceptibility for the development of atherosclerosis, bred into PLTP knockout mice. The data clearly show that PLTP can be proatherogenic. As mentioned above, however, PLTP may have antiatherogenic potential in wild-type C57BL6 mice. Information regarding the role and regulation of PLTP in human (patho)physiology is still relatively sparse but accumulating rapidly. PLTP activity is elevated in diabetes mellitus (both type 1 and type 2), obesity and insulin resistance. PMID- 11891416 TI - Cytoplasmic fatty acid-binding proteins: emerging roles in metabolism and atherosclerosis. AB - Cytoplasmic fatty acid-binding proteins (FABPs) are a family of proteins, expressed in a tissue-specific manner, that bind fatty acid ligands and are involved in shuttling fatty acids to cellular compartments, modulating intracellular lipid metabolism, and regulating gene expression. Several members of the FABP family have been shown to have important roles in regulating metabolism and have links to the development of insulin resistance and the metabolic syndrome. Recent studies demonstrate a role for intestinal FABP in the control of dietary fatty acid absorption and chylomicron secretion. Heart FABP is essential for normal myocardial fatty acid oxidation and modulates fatty acid uptake in skeletal muscle. Liver FABP is directly involved in fatty acid ligand signaling to the nucleus and interacts with peroxisome proliferator-activated receptors in hepatocytes. The adipocyte FABP (aP2) has been shown to affect insulin sensitivity, lipid metabolism and lipolysis, and has recently been shown to play an important role in atherosclerosis. Interestingly, expression of aP2 by the macrophage promotes atherogenesis, thus providing a link between insulin resistance, intracellular fatty acid disposition, and foam cell formation. The FABPs are promising targets for the treatment of dyslipidemia, insulin resistance, and atherosclerosis in humans. PMID- 11891417 TI - Gene:environment interaction in lipid metabolism and effect on coronary heart disease risk. AB - Both genetic and environmental factors influence coronary heart disease, therefore studies of coronary heart disease risk are often confounded by gene:gene and gene:environment interactions. Such interactions imply that at the molecular level there is synergy between the gene products or with the by products of the environmental insult, resulting in a greater than additive effect on risk. Genetic risk is thus modifiable in an environment-specific manner. This review focuses on recently reported effects of smoking (environmental factor) on the impact of variation in the genes for glutathione S-transferase, paraoxonase and apolipoprotein E on the risk of coronary heart disease and effects on intermediate lipid traits. We end on a cautionary note for the need for repeat studies to confirm these reported gene:environment effects. PMID- 11891418 TI - Dietary polyunsaturated fatty acids and regulation of gene transcription. AB - Dietary polyunsaturated fatty acids (PUFAs) are a source of energy and structural components for cells. PUFAs also have dramatic effects on gene expression by regulating the activity or abundance of four families of transcription factor, including peroxisome proliferator activated receptor (PPAR) (alpha, beta and gamma), liver X receptors (LXRs) (alpha and beta), hepatic nuclear factor-4 (HNF 4)alpha and sterol regulatory element binding proteins (SREBPs) 1 and 2. These transcription factors play a major role in hepatic carbohydrate, fatty acid, triglyceride, cholesterol and bile acid metabolism. Non-esterified fatty acids or fatty acid metabolites bind to and regulate the activity of PPARs, LXRs and HNF 4. In contrast, PUFAs regulate the nuclear abundance of SREBPs by controlling the proteolytic processing of SREBP precursors, or regulating transcription of the SREBP-1c gene or turnover of mRNA(SREBP-1c). The n3 and n6 PUFAs are feed-forward activators of PPARs, while these same fatty acids are feedback inhibitors of LXRs and SREBPs. Saturated fatty acyl coenzyme A thioesters activate HNF-4 alpha, while coenzyme A thioesters of PUFAs antagonize HNF-4 alpha action. Understanding how fatty acids regulate the activity and abundance of these and other transcription factors will likely provide insight into the development of novel therapeutic strategies for better management of whole body lipid and cholesterol metabolism. PMID- 11891419 TI - Phenotypic variability in familial hypercholesterolaemia: an update. AB - Heterozygous familial hypercholesterolaemia is among the most common inherited dominant disorders, and is characterized by severely elevated LDL-cholesterol levels and premature cardiovascular disease. Although the cause of familial hypercholesterolaemia is monogenic, there is a substantial variation in the onset and severity of atherosclerotic disease symptoms. Additional atherogenic risk factors of environmental, metabolic and genetic origin, in conjunction with the LDL receptor defect, are presumed to influence the clinical phenotype in familial hypercholesterolaemia. The present review discusses recent developments in this field. PMID- 11891420 TI - Expression profiling and comparative sequence derived insights into lipid metabolism. AB - Expression profiling and genomic DNA sequence comparisons are increasingly being applied to the identification and analysis of the genes that are involved in lipid metabolism. Not only has genome-wide expression profiling aided in the identification of novel genes that are involved in important processes in lipid metabolism such as sterol efflux, but also the utilization of information from these studies has added to our understanding of the regulation of pathways that participate in the process. Coupled with these gene expression studies, cross species comparison (a technique used to search for sequences that are conserved through evolution) has proven to be a powerful tool to identify important noncoding regulatory sequences and novel genes that are relevant to lipid biology. An example of the value of this approach was the recent chance discovery of a new apolipoprotein gene (that which encodes apolipoprotein AV) that has dramatic effects on triglyceride metabolism in mice and humans. PMID- 11891421 TI - Understanding atherosclerosis through mouse genetics. AB - During the past year studies with mouse models have significantly clarified our understanding of atherosclerosis. Noteworthy achievements include: the discovery of a number of novel genes and pathways; new evidence emphasizing the role of lymphocytes in atherogenesis; the development of mouse models exhibiting advanced lesions with evidence of thrombosis; and new results indicating an anti atherogenic effect of testosterone. PMID- 11891422 TI - New mouse models for lipoprotein metabolism and atherosclerosis. AB - Transgenic mouse models have been crucial to our current understanding of the mechanisms of lipoprotein metabolism. Moreover, these models have greatly advanced our understanding of the pathology associated with altered lipoprotein levels. Recent progress has been made in cellular uptake, intracellular metabolism, cellular efflux mechanisms and transcriptional regulation. In particular, much progress has been made in our understanding of events that take place in the vessel wall. In addition, the transgenic mouse model is becoming a crucial tool in genomic studies to evaluate gene function, as well as a subject of genome-wide expression studies. The present review describes progress in all of these areas and shows that animal models are likely to remain important to our view of gene function in the context of the whole organism. PMID- 11891423 TI - Nutrition and metabolism. PMID- 11891424 TI - Genetics and molecular biology. PMID- 11891425 TI - Lipid metabolism: peroxisome proliferator-activated receptor-alpha and atherosclerosis. PMID- 11891426 TI - Hyperlipidaemia and cardiovascular disease. PMID- 11891427 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 11891428 TI - Therapy and clinical trials. PMID- 11891429 TI - Vascular genomics of the human brain. AB - The microvasculature of the human brain plays an important role in the development and maintenance of the central nervous system and in the pathogenesis of brain diseases, and is the site of differential gene expression within the brain. However, human brain microvascular-specific genes may not be detected in whole-brain gene microarray because the volume of the brain microvascular endothelium is relatively small (0.1%) compared with the whole brain. Therefore, the differential gene expression within the human brain microvasculature was evaluated using suppression subtractive hybridization with RNA isolated from human brain microvessels. Gene identification was restricted to the first 71 clones that were differentially expressed at the brain microvasculature. Twenty of these were genes encoding proteins with known function that were involved in angiogenesis, neurogenesis, molecular transport, and maintenance of endothelial tight junctions or the cytoskeleton. Eighteen genes coding for proteins of an unknown function were identified, including five genes containing satellite DNA sequences. The results provide the initial outline of the genomics of the human brain microvasculature, and have implications for the identification of both targets for brain-specific drug transport and changes in microvascular gene expression in brain diseases. PMID- 11891430 TI - Late reversal of cerebral perfusion and water diffusion after transient focal ischemia in rats. AB - Region-specific cerebral blood flow (CBF) and the apparent diffusion coefficient (ADC) of tissue water in the rat brain were quantified by high-field magnetic resonance imaging at 9.4 T in the rat suture occlusion model. Cerebral blood flow and ADC were compared during the short- (4.5 hours) and long-term (up to 6 days) reperfusion after 80 minutes of transient middle cerebral artery occlusion, and correlated with the histology analysis. On occlusion, average CBF fell from approximately 100 to less than 50 mL x 100 g(-1) x min(-1) in the cortex, and to less than 20 mL x 100 g(-1) x min(-1) in the caudate putamen (CP). Corresponding ADC values decreased from (6.98 +/- 0.82) x 10(-4) to (5.49 +/- 0.54) x 10(-4) mm2/s in the cortex, and from (7.16 +/- 0.58) x 10(-4) to (4.86 +/- 0.62) x 10( 4) mm2/s in the CP. On average, CBF recovered to approximately 50% of baseline in the first 24 hours of reperfusion. After 2 to 4 days, a strong hyperperfusion in the ipsilateral cortex and CP, up to approximately 300 mL x 100 g(-1) x min(-1), was observed. The ADC ratio in the ipsilateral and contralateral CP was also inverted in the late reperfusion period. Histology revealed more severe tissue damage at the late stage of reperfusion than at 4.5 hours. Significant reversal of CBF and ADC during the late reperfusion period may reflect the impairment of autoregulation in the ischemic regions. Vascular factors may play an important role in the infarct development after 80-minute focal ischemia. PMID- 11891431 TI - Quantitative assessment of the balance between oxygen delivery and consumption in the rat brain after transient ischemia with T2 -BOLD magnetic resonance imaging. AB - The balance between oxygen consumption and delivery in the rat brain after exposure to transient ischemia was quantitatively studied with single-spin echo T2-BOLD (blood oxygenation level-dependent) magnetic resonance imaging at 4.7 T. The rats were exposed to graded common carotid artery occlusions using a modification of the four-vessel model of Pulsinelli. T2, diffusion, and cerebral blood volume were quantified with magnetic resonance imaging, and CBF was measured with the hydrogen clearance method. A transient common carotid artery occlusion below the CBF value of approximately 20 mL x 100 g(-1) x min(-1) was needed to yield a T2 increase of 4.6 +/- 1.2 milliseconds (approximately 9% of cerebral T2) and 6.8 +/- 1.7 milliseconds (approximately 13% of cerebral T2) after 7 and 15 minutes of ischemia, respectively. Increases in CBF of 103 +/- 75% and in cerebral blood volume of 29 +/- 20% were detected in the reperfusion phase. These hemodynamic changes alone could account for only approximately one third of the T2 increase in luxury perfusion, suggesting that a substantial increase in blood oxygen saturation (resulting from reduced oxygen extraction by the brain) is needed to explain the magnetic resonance imaging observation. PMID- 11891432 TI - Striking differences in glucose and lactate levels between brain extracellular fluid and plasma in conscious human subjects: effects of hyperglycemia and hypoglycemia. AB - Brain levels of glucose and lactate in the extracellular fluid (ECF), which reflects the environment to which neurons are exposed, have never been studied in humans under conditions of varying glycemia. The authors used intracerebral microdialysis in conscious human subjects undergoing electrophysiologic evaluation for medically intractable epilepsy and measured ECF levels of glucose and lactate under basal conditions and during a hyperglycemia-hypoglycemia clamp study. Only measurements from nonepileptogenic areas were included. Under basal conditions, the authors found the metabolic milieu in the brain to be strikingly different from that in the circulation. In contrast to plasma, lactate levels in brain ECF were threefold higher than glucose. Results from complementary studies in rats were consistent with the human data. During the hyperglycemia hypoglycemia clamp study the relationship between plasma and brain ECF levels of glucose remained similar, but changes in brain ECF glucose lagged approximately 30 minutes behind changes in plasma. The data demonstrate that the brain is exposed to substantially lower levels of glucose and higher levels of lactate than those in plasma; moreover, the brain appears to be a site of significant anaerobic glycolysis, raising the possibility that glucose-derived lactate is an important fuel for the brain. PMID- 11891433 TI - Induction of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in rat brain after focal ischemia/reperfusion. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme, has been recently identified to be involved in the initiation of neuronal apoptosis. To investigate the serial changes and cellular localization of GAPDH expression, and its role in ischemia/reperfusion-induced neuronal apoptosis, the authors analyzed immunohistochemically brain areas of rats subjected to middle cerebral artery occlusion (MCAO) and reperfusion. Nuclear overexpression of GAPDH was noted in the ischemic core area after 2 hours of MCAO without reperfusion. During the subsequent reperfusion, nuclear accumulation of GAPDH in this area decreased in a time-dependent manner. However, cytoplasmic and nuclear GAPDH immunoreactivity was detected in neurons of the penumbra area of the parietal cortex, in rats subjected to 2-hour MCAO followed by 3-hour reperfusion. The increase of nuclear GAPDH immunoreactivity was persistently noted up to 48 hours of reperfusion, whereas cytoplasmic immunoreactivity correlated inversely with the duration of reperfusion. Moreover, double staining revealed colocalization of nuclear GAPDH and TUNEL in the penumbra area. The authors' study demonstrated that overexpression of GAPDH and nuclear translocation occurred in both the ischemic core and penumbra area soon after focal ischemia. These processes could be viewed as an early marker of ischemia/reperfusion-induced apoptotic neuronal death. The results suggest that GAPDH may play a critical role in the progression and spread of ischemic neuronal damage. PMID- 11891434 TI - High sensitivity of protoplasmic cortical astroglia to focal ischemia. AB - SUMMARY: The generally accepted concept that astrocytes are highly resistant to hypoxic/ischemic conditions has been challenged by an increasing amount of data. Considering the differences in functional implications of protoplasmic versus fibrous astrocytes, the authors have investigated the possibility that those discrepancies come from specific behaviors of the two cell types. The reactivity and fate of protoplasmic and fibrous astrocytes were observed after permanent occlusion of the medial cerebral artery in mice. A specific loss of glial fibrillary acidic protein (GFAP) immunolabeling in protoplasmic astrocytes occurred within minutes in the area with total depletion of regional CBF (rCBF) levels, whereas "classical" astrogliosis was observed in areas with remaining rCBF. Severe disturbance of cell function, as suggested by decreased GFAP content and increased permeability of the blood-brain barrier to macromolecules, was rapidly followed by necrotic cell death, as assessed by ultrastructure and by the lack of activation of the apoptotic protease caspase-3. In contrast to the response of protoplasmic astrocytes, fibrous astrocytes located at the brain surface and in deep cortical layers displayed a transient and limited hypertrophy, with no conspicuous cell death. These results point to a differential sensitivity of protoplasmic versus fibrous cortical astrocytes to blood deprivation, with a rapid demise of the former, adding to the suggestion that protoplasmic astrocytes play a crucial role in the pathogenesis of ischemic injury. PMID- 11891435 TI - Increased proliferation of neural progenitor cells but reduced survival of newborn cells in the contralateral hippocampus after focal cerebral ischemia in rats. AB - Recent studies demonstrated that neurogenesis in the adult hippocampus increased after transient global ischemia; however, the molecular mechanism underlying increased neurogenesis after ischemia remains unclear. The finding that proliferation of progenitor cells occurred at least a week after ischemic insult suggests that the stimulus was not an ischemic insult to progenitor cells. To clarify whether focal ischemia increases the rate of neurogenesis in the remote area, the authors examined the contralateral hemisphere in rats subjected to permanent occlusion of the middle cerebral artery. In the subgranular zone of the hippocampal dentate gyrus, the numbers of bromodeoxyuridine (BrdU)-positive cells increased approximately sixfold 7 days after ischemia. In double immunofluorescence staining, more than 80% of newborn cells expressed Musashi1, a marker of neural stem/progenitor cells, but only approximately 10% of BrdU positive cells expressed glial fibrillary acidic protein (GFAP), a marker of astrocytes. The number of BrdU-positive cells markedly decreased 28 days after BrdU administration after ischemia, but it was still elevated compared with that of sham-operated rats. In double immunofluorescence staining, 80% of newborn cells expressed NeuN, a marker of differentiated neurons, and 10% of BrdU positive cells expressed GFAP. However, in the other areas of the contralateral hemisphere including the rostral subventricular zone, the number of BrdU-positive cells remained unchanged. These results showed that focal ischemia stimulated the proliferation of neuronal progenitor cells, but did not support survival of newborn cells in the contralateral hippocampus. PMID- 11891436 TI - Monocyte chemoattractant protein-1 deficiency is protective in a murine stroke model. AB - Inflammatory processes have been implicated in the pathogenesis of brain damage after stroke. In rodent stroke models, focal ischemia induces several proinflammatory chemokines, including monocyte chemoattractant protein-1 (MCP-1). The individual contribution to ischemic tissue damage, however, is largely unknown. To address this question, the authors subjected MCP-1-deficient mice (MCP-1-/-) to permanent middle cerebral artery occlusion (MCAO). Measurement of basal blood pressure, cerebral blood flow, and blood volume revealed no differences between wild-type (wt) and MCP-1-/- mice. MCAO led to similar cerebral perfusion deficits in wt and MCP-1-/- mice, excluding differences in the MCA supply territory and collaterals. However, compared with wt mice, the mean infarct volume was 29% smaller in MCP-1-/- mice 24 hours after MCAO (P = 0.022). Immunostaining showed a reduction of phagocytic macrophage accumulation within infarcts and the infarct border in MCP-1-/- mice 2 weeks after MCAO. At the same time point, the authors found an attenuation of astrocytic hypertrophy in the infarct border and thalamus in MCP-1-/- mice. However, these effects on macrophages and astrocytes in MCP-1-/- mice occurred too late to suggest a protective role in acute infarct growth. Of note: at 6 hours after MCAO, MCP-1-/- mice produced significantly less interleukin-1beta in ischemic tissue; this might be related to tissue protection. The results of this study indicate that inhibition of MCP-1 signaling could be a new acute treatment approach to limit infarct size after stroke. PMID- 11891437 TI - The systemic and local acute phase response following acute brain injury. AB - It is not known whether acute brain injury results in a systemic acute phase response (APR) or whether an APR influences outcome after an insult to the CNS. The present study sought to establish whether brain injury elicits a systemic or local APR. The expression of acute phase protein (APP) mRNA in liver and brain tissues was measured by Taqman reverse transcriptase-polymerase chain reaction after an excitotoxic lesion in the striatum or challenge with a proinflammatory cytokine. N-methyl-d-aspartate (NMDA)-induced brain lesion did not elicit a systemic APR. In contrast, proinflammatory challenge with mouse recombinant interleukin-1beta (mrIL-1beta) resulted in a significant hepatic APP mRNA expression within 6 hours. Thus, an inflammatory challenge that results in a meningitis leads to a hepatic APR, whereas acute brain injury alone, with no evidence of a meningitis, does not produce an APR. This is surprising because NMDA leads to an increase in endogenous IL-1beta synthesis. This suggests that the brain has an endogenous antiinflammatory mechanism, which protects against the spread of inflammation after an acute injury. In the brain, both excitotoxic lesions and proinflammatory challenge resulted in a profound parenchymal upregulation of APP mRNA after 6 and 12 hours in the injected hemisphere. These results suggest that the local APR may play a role as an antiinflammatory mechanism. These findings indicate a potentially pivotal role for peripheral and local APP production on outcome after brain injury. PMID- 11891438 TI - Differential activation of mitogen-activated protein kinase pathways after traumatic brain injury in the rat hippocampus. AB - Mitogen-activated protein kinases, which play a crucial role in signal transduction, are activated by phosphorylation in response to a variety of mitogenic signals. In the present study, the authors used Western blot analysis and immunohistochemistry to show that phosphorylated extracellular signal regulated protein kinase (p-ERK) and c-Jun NH2-terminal kinase (p-JNK), but not p38 mitogen-activated protein kinase, significantly increased in both the neurons and astrocytes after traumatic brain injury in the rat hippocampus. Different immunoreactivities of p-ERK and p-JNK were observed in the pyramidal cell layers and dentate hilar cells immediately after traumatic brain injury. Immunoreactivity for p-JNK was uniformly induced but was only transiently induced throughout all pyramidal cell layers. However, strong immunoreactivity for p-ERK was observed in the dentate hilar cells and the damaged CA3 neurons, along with the appearance of pyknotic morphologic changes. In addition, immunoreactivity for p-ERK was seen in astrocytes surrounding dentate and CA3 pyramidal neurons 6 hours after traumatic brain injury. These findings suggest that ERK and JNK but not p38 cascades may be closely involved in signal transduction in the rat hippocampus after traumatic brain injury. PMID- 11891439 TI - Near-infrared spectroscopy cerebral oxygen saturation thresholds for hypoxia ischemia in piglets. AB - Detection of cerebral hypoxia-ischemia remains problematic in neonates. Near infrared spectroscopy, a noninvasive bedside technology has potential, although thresholds for cerebral hypoxia-ischemia have not been defined. This study determined hypoxic-ischemic thresholds for cerebral oxygen saturation (SCO2) in terms of EEG, brain ATP, and lactate concentrations, and compared these values with CBF and sagittal sinus oxygen saturation (SVO2). Sixty anesthetized piglets were equipped with near-infrared spectroscopy, EEG, laser-Doppler flowmetry, and a sagittal sinus catheter. After baseline, SCO2 levels of less than 20%, 20% to 29%, 30% to 39%, 40% to 49%, 50% to 59%, 60% to 79%, or 80% or greater were recorded for 30 minutes of normoxic normocapnia, hypercapnic hyperoxia, or bilateral carotid occlusion with or without arterial hypoxia. Brain ATP and lactate concentrations were measured biochemically. Logistic and linear regression determined the SCO2, CBF, and SVO2 thresholds for abnormal EEG, ATP, and lactate findings. Baseline SCO2 was 68 + 5%. The SCO2 thresholds for increased lactate, minor and major EEG change, and decreased ATP were 44 +/- 1%, 42 +/- 5%, 37 +/- 1%, and 33 +/- 1%. The SCO2 correlated linearly with SVO2 (r = 0.98) and CBF (r = 0.89), with corresponding SVO2 thresholds of 23%, 20%, 13%, and 8%, and CBF thresholds (% baseline) of 56%, 52%, 42%, and 36%. Thus, cerebral hypoxia-ischemia near-infrared spectroscopy thresholds for functional impairment are SCO2 33% to 44%, a range that is well below baseline SCO2 of 68%, suggesting a buffer between normal and dysfunction that also exists for CBF and SVO2. PMID- 11891440 TI - Comparison of the effects of cyclosporin a on the metabolism of perfused rat brain slices during normoxia and hypoxia. AB - The authors evaluated and compared the metabolic effects of cyclosporin A in the rat brain during normoxia and hypoxia/reperfusion. Ex vivo 31P magnetic resonance spectroscopy experiments based on perfused rat brain slices showed that under normoxic conditions, 500 microg/L cyclosporin A significantly reduced mitochondrial energy metabolism (nucleotide triphosphate, 83 +/- 9% of controls; phosphocreatine, 69 +/- 9%) by inhibition of the Krebs cycle (glutamate, 77 +/- 5%) and oxidative phosphorylation (NAD+, 65 +/- 14%) associated with an increased generation of reactive oxygen species (285 +/- 78% of control). However, the same cyclosporin A concentration (500 microg/L) was found to be the most efficient concentration to inhibit the hypoxia-induced mitochondrial release of Ca2+ in primary rat hippocampal cells with cytosolic Ca2+ concentrations not significantly different from normoxic controls. Addition of 500 microg/L cyclosporin A to the perfusion medium protected high-energy phosphate metabolism (nucleotide triphosphate, 11 +/- 15% of control vs. 35 +/- 9% with 500 microg/L cyclosporin A) and the intracellular pH (6.2 +/- 0.1 control vs. 6.6 +/- 0.1 with cyclosporin A) in rat brain slices during 30 minutes of hypoxia. Results indicate that cyclosporin A simultaneously decreases and protects cell glucose and energy metabolism. Whether the overall effect was a reduction or protection of cell energy metabolism depended on the concentrations of both oxygen and cyclosporin A in the buffer solution. PMID- 11891441 TI - Spatial integration of vascular changes with neural activity in mouse cortex. AB - The authors evaluated representations of discretely activated, neighboring brain regions using real-time optical intrinsic signals by transcranial imaging with 540-nm and 610-nm broadband illumination of the mouse barrel cortex. Iron filings were glued to two neighboring whiskers (C2 + D2) that were stimulated magnetically, singly and together. Real-time images were collected, averaged, and analyzed statistically. Postmortem filling of arteries with fluorescent beads was shown in relation to histochemical staining of barrels to accurately relate surface changes to functional cortical columns. Significant optical intrinsic signal changes are related to overlapping distributions of arterioles that feed the two separate areas. Activation of adjacent and interacting cortical columns leads not only to increased magnitude of vascular responses in those columns, but also to wider spatial extent of absorption changes occurring principally in areas of cortex fed by vessels upstream of the active cortex. The localization of changing hemoglobin absorption around upstream blood vessels and their vascular domains suggests that propagated vasodilation of upstream parent vessels is greater when vasodilatory signals from separate areas of active cortex converge on common arterioles that feed them. PMID- 11891442 TI - A reinvestigation of the extended counting method for fractal analysis of the pial vasculature. PMID- 11891445 TI - Persistent low back pain and sciatica in the United States: treatment outcomes. AB - Patients with persistent low back pain (LBP) appear to be different in several important ways from patients who have traditionally been classified as patients with acute or chronic LBP, and data on the effectiveness of the treatments prescribed for them are lacking. The aim of the current study was to evaluate the short- and long-term effectiveness of the treatments currently prescribed for these patients. The data reported in this article were gathered as part of a multicenter, prospective, cross-sectional study of patients who were treated for persistent LBP by neurologic and orthopedic surgeons who are recognized specialists in spinal disorders. At enrollment, patients completed a baseline evaluation, and their physicians recorded relevant clinical and treatment data on standardized study forms. At 3, 6, 12, and 24 months after treatment, patients completed follow-up evaluations. Patients were divided into five treatment groups, and effectiveness was evaluated separately for each group using five patient-reported measures of outcome: pain severity, functional disability, psychologic distress, physical symptoms, and health care use. The data revealed that at the 2-year follow-up, the typical patient of the no-treatment group had improved slightly in terms of pain severity and health care use, but had experienced little or no improvement in functional disability, physical symptoms, and psychologic distress. The average patient in the conservative care group reported small improvements in pain severity, functional disability, physical symptoms, and health care use, with no change in psychologic distress. These small improvements occurred within the first 3 months after enrollment, with essentially no change thereafter. The average patient in the immediate surgical care group showed substantial improvement on all of the outcome measures. The observed improvements were evident shortly after treatment and were maintained for the duration of the study. Patients in the delayed surgical care group had outcomes that were less dramatic than those observed in the immediate surgery care group, but greater than those observed in the conservative care group. The patients who were treated surgically by physicians outside the study, outside surgical care group, did not improve over time. Patients with persistent LBP who received no treatment showed no spontaneous recovery. Conservative care treatments prescribed by surgeons who specialize in spinal disorders, did not appear to be any more effective than no treatment. The outcome of surgery for persistent LBP varied from dramatic for one subgroup of surgical patients, to poor for another subgroup of patients. Patients who were selected immediately for surgical treatment improved substantially. Those treated surgically later by study physicians or by physicians not associated with the study fared less well. PMID- 11891447 TI - The association of sagittal spinal and pelvic parameters in asymptomatic persons and patients with isthmic spondylolisthesis. AB - Using a specialized orthopedic software package, the authors investigated the sagittal spinal shape and the position of the pelvis in the space in patients with isthmic spondylolisthesis and in persons with no such symptoms. Digitized lateral spinal radiographs of 30 healthy volunteers and 48 patients were evaluated. The absolute values and significant correlations between parameters were analyzed. The pelvic parameters correlated well with lordosis, which shows sagittal balance in the asymptomatic group. The hyperlordosis and the horizontally positioned sacrum in isthmic spondylolisthesis enlarge the tensile force component of gravity, which may cause the lysis. Finally, the authors developed a new balance between the pelvis and the spine after slipping of the vertebral body. The degree of slipping correlated well with the sacrofemoral anatomic constant (incidence), which is unique in each person. PMID- 11891446 TI - Moving toward a standard for spinal fusion outcomes assessment. AB - Previous spinal fusion outcomes assessment studies have been complicated by inconsistencies in evaluative criteria and consequent variations in results. As a result, a general consensus is lacking on how to achieve comprehensive outcomes assessment for spinal fusion surgeries. The purpose of this article is to report the most validated and frequently used assessment measures to facilitate comparable outcomes studies in the future. Twenty-seven spinal fusion outcomes studies published between 1990 and 2000 were retrospectively reviewed. Study characteristics such as design, evaluative measures, and assessment tools were recorded and analyzed. Based on the reviewed literature, an outcomes assessment model is proposed including the Short Form-36 Health Survey, the Oswestry Disability Questionnaire, the North American Spine Society Patient Satisfaction Index, the Prolo Economic Scale, a 0-10 analog pain scale, medication use, radiographically assessed fusion status, and a generalized complication rate. PMID- 11891448 TI - Unilateral transforaminal posterior lumbar interbody fusion (TLIF): indications, technique, and 2-year results. AB - A prospective analysis of consecutive cases of lumbar fusion using the unilateral transforaminal posterior lumbar interbody fusion (TLIF) technique with pedicle screw fixation. The objective of the study was to assess the clinical and radiographic outcome of TLIF and describe the technique and indications in the treatment of degenerative disease of the lumbar spine. Forty patients treated with TLIF for degenerative diseases of the lumbar spine were followed up for a minimum of 2.5 years (mean: 36 months; range: 30-42 months). Twenty-three patients had degenerative disc disease alone, 13 had associated isthmic or degenerative spondylolisthesis, and 4 had recurrent disc herniations at the L4-L5 level. Thirty-six (90%) had solid fusions radiographically at latest follow-up. Seventy-nine percent had excellent or good clinical outcomes. Our patients demonstrated high fusion rates and patient satisfaction. PMID- 11891449 TI - Unilateral laminectomy for bilateral decompression of lumbar spinal stenosis: a prospective comparative study with conservatively treated patients. AB - The authors performed single- or multiple-level unilateral laminectomy to treat lumbar spinal stenosis in patients with mild to moderate leg pain and compared the results with those from patients treated with conservative therapy in a prospective study. This decompression technique produced a 68% rate of improvement compared with a 33% rate for conservatively treated patients. The surgical group exhibited significant and sustained improvement, whereas the functional and clinical status of the conservatively managed group had returned to baseline during the same period. The preoperative dural sac cross-sectional area at the level of the most stenosis was 70.76 +/- 28.2 mm(2) for the surgical group, whereas on postoperative scans it was 108.12 +/- 31.5 mm(2), with an average correction rate of 65%. Neither new degenerative spondylolisthesis nor any evidence of instability was detected in any patient during the study. PMID- 11891451 TI - Preoperative CT examination for accurate and safe anterior spinal instrumentation surgery with endoscopic approach. AB - The purpose of this article is to introduce a new procedure for the surgical planning of thoracic anterior spinal instrumentation via endoscopy. For accurate and safe anterior screw insertion via the endoscopic approach, we devised a surgical plan based on the preoperative chest computed tomography (CT) findings obtained with radiographic markers. Using this method, we performed endoscopic thoracic spinal instrumentation surgery in 14 patients. Nine patients underwent anterior endoscopic correction and fusion of idiopathic scoliosis by Cotrel Dubousset instrumentation, and five patients underwent anterior endoscopic spinal fixation with instrumentation. The accuracy of screw insertion was evaluated postoperatively by CT scanning. One interbody fusion cage and 53 screws were inserted in the 14 patients using endoscopy. Postoperative CT scans revealed that the screws were all accurately inserted without any neurologic complications. In conclusion, using this novel procedure for surgical planning based on CT findings obtained with radiographic markers, anterior screws can be inserted safely and accurately via an endoscopic approach. PMID- 11891453 TI - A biomechanical comparison of posterolateral fusion and posterior fusion in the lumbar spine. AB - Late postoperative complications occurred after posterior fusion and posterolateral fusion as a result of biomechanical alterations. The stress change between the two fusion procedures has not been well reported. To differentiate the biomechanical alteration that occurs with posterior fusion and posterolateral fusion of the lumbar spine, the load sharing of the vertebrae, disc, facet joint, bone graft, and the range of motion were computed in a finite element model. Five finite element models, including the intact lumber spine, posterior fusion, posterior fusion with implant, posterolateral fusion, and posterolateral fusion with implant, were created for stress analysis. The finite element model estimated that the differences between these two fusion procedures were within 7% in stress of the adjacent disc, 3% in force of the facet joint above the fusion mass, and 5% in the range of motion. However, the stress of the pedicle in posterolateral fusion without an implant was at most two times greater than that in the intact lumbar spine under lateral bending. The stress of pars interarticularis in posterior fusion without an implant was also at most two times greater than that in the intact lumbar spine under lateral bending. After the implant was added, the discrepancy between the two fusion procedures decreased but still remained a relatively large difference. Therefore, the largest changes of posterior fusion and posterolateral fusion were in the pars interarticularis and pedicle, respectively. PMID- 11891455 TI - Biomechanical evaluation of a double-threaded pedicle screw in elderly vertebrae. AB - We sought to test the hypothesis that a pedicle screw that has two parallel threads of different heights throughout the full length of the screw could increase both bone purchase and pull-out strength compared with a standard single threaded screw of similar dimensions. A single-threaded pedicle screw and a double-threaded pedicle screw were respectively placed into the paired pedicles of 21 vertebral bodies. The screws were then pulled out of the pedicles, and output parameters were measured. Although insertional torque was, on average, 14.5% higher (p = 0.039) for the single-threaded screw, maximum pull-out strength (p = 0.12), energy-to-failure (p = 0.39), and stiffness (p = 0.54) were not statistically different for the two screw types. It is concluded that a second, smaller inner thread on a double-threaded pedicle screw does not translate into either increased bone purchase or higher pull-out strengths. PMID- 11891458 TI - Possible complications of anterior perforation of the vertebral body using cervical pedicle screws. AB - No previous studies have analyzed the possible complications of anterior perforation of the cervical vertebral body with pedicle screws. The objective of this study was to identify the possible implications of an anterior vertebral body perforation. Ten consecutive Euro-American cadavers (C2-C7) were used. The male-to-female ratio was 3:7. The average specimen age was 79.6 years (range: 65 97 years), and average height was 159 cm (range: 155-175 cm). Axial computed tomography scans through the isthmus of pedicles were taken. Five millimeter and 10 mm margins anterior to the vertebral bodies were defined. Within 5 mm anterior to the anterior cortex of the vertebral body, we found mostly muscles (at C2: m. longus colli and pharyngeal constrictors; at C3 and C4: m. scalenus medius, longus colli, pharyngopalatinus and pharyngeal constrictors; at C5 and C6: m. longus colli and longus capitis; and at C7: m. longus colli), except at C3, C4, and C7, where the pharynx and esophagus were within the margin. Between 6 and 10 mm, we found mostly hollow organs (at C2: pharynx and small veins; at C3 and C4: the same muscles as within the 5 mm margin, with addition of the pharynx and some small veins; at C5 and C6: pharynx, pharyngeal constrictors and the thyroid cartilage; and at C7: the esophagus). Except C2, there is no safe zone anterior to the cervical vertebral bodies in the cervical spine, which would allow bicortical purchase of pedicle screws without being close to important surrounding structures. PMID- 11891459 TI - Progressive spinal lordosis after laminoplasty in a child with thoracic neuroblastoma. AB - Laminoplasty has been advocated increasingly after spinal tumor excision in children. Results have shown that it offers the required decompression, while maintaining spinal stability and the integrity of the posterior vertebral elements. To the authors' knowledge, there has been no description of a progressive lordotic deformity of the thoracic spine after this procedure. A case of an 8-year-old boy with thoracic neuroblastoma developing progressive thoracic lordosis after laminoplasty is reviewed, and a possible cause is suggested. Discussing this potential complication with parents and the patient, and following up with regular clinical and radiographic assessments is advised. PMID- 11891456 TI - Does anterior plating maintain cervical lordosis versus conventional fusion techniques? A retrospective analysis of patients receiving single-level fusions. AB - A retrospective review of medical records and radiographs of patients receiving anterior cervical discectomy and fusion (ACDF) without anterior plating and with anterior plating was performed. The objective of the study was to determine whether a difference exists in cervical lordotic alignment between subjects undergoing single-level ACDF with and without anterior cervical plating instrumentation for symptomatic cervical disc disease. Collapse or settling of grafted bone into the vertebral endplates with resulting kyphotic deformity of the cervical spine is a commonly described complication of anterior discectomy and fusion. Despite the increasing use of instrumentation for the treatment of cervical spine injuries and degenerative conditions, little is known regarding lordotic alignment of the cervical spine in patients who receive plating instrumentation compared with conventional fusion without plating. Accumulating evidence suggests that plating is superior to non-plating techniques in patients with multiple level cervical disc lesions in regard to fusion, return to work rates, and complication rates; however, little is known about maintenance of lordotic curve alignment in single- and multiple-level procedures. Neutral lateral cervical radiographs of 57 patients who underwent single-level ACDF between 1994 and 1999 with anterior screw plates (n = 26), and conventional single-level fusion without anterior screw plates (n = 21) were retrospectively assessed. Measurements were made on weight-bearing lateral cervical radiographs to assess overall sagittal spinal alignment and intersegmental sagittal alignment at the surgical site before surgery, immediately after surgery, 4 to 12 weeks after surgery, and 12+ months after surgery. The average magnitude of overall lordosis measured between C2 and C7 decreased 4.2 degrees in the non-plated group, while being preserved in the plated group. This finding did not reach statistical significance in the long-term follow-up. At the surgical site, the segmental contribution to lordosis decreased an average 2.5 degrees in the non plated group versus an increase of 5.67 degrees in the plated group, and this finding was statistically significant between groups measured at all pre- and postoperative visits (p < 0.01). On average, the plating procedure resulted in preserving overall lordosis while increasing the magnitude of segmental lordosis at the surgical site. In comparison, the conventional method resulted in a net loss of overall lordosis and segmental lordosis at the surgical site. PMID- 11891461 TI - Congenital anomaly of craniovertebral junction: atlas-dens fusion with C1 anterior arch cleft. AB - A rare case of congenital atlantoaxial block is reported. A 13-year-old boy had a fusion of a non-separated odontoid process with the anterior arch of the atlas, in association with an anterior midline C1 arch cleft. PMID- 11891462 TI - Intramedullary tuberculoma with syringomyelia. AB - Intramedullary tuberculoma with syringomyelia is rare. We treated a woman with back pain and weakness of the left leg that had slowly progressed for more than 30 years. Radiologic evaluation demonstrated a crescent-shaped calcification at the level of the C6 vertebra, and syringomyelia from C7 to T9. Laminectomy and syringosubdural shunt placement were performed, and a tuberculoma was removed. Back pain resolved after the operation, and mobility was facilitated. We recommend surgery for intrathecal tuberculoma with syringomyelia even when the course has been prolonged with no active tuberculous lesion. PMID- 11891463 TI - Current status and future applications of cardiac receptor imaging with positron emission tomography. PMID- 11891464 TI - The role of somatostatin receptor scintigraphy in the management of pituitary tumours. AB - The detection of functioning neuroendocrine tissue bearing somatostatin receptors has been facilitated to a large extent by the availability of radiolabelled octreotide scanning with 111In octreotide. This review discusses the possible role for somatostatin receptor scintigraphy (SRS) in the evaluation of pituitary adenomas. PMID- 11891465 TI - Initial evaluation of the feasibility of single photon emission tomography with p [123 I]iodo-L-phenylalanine for routine brain tumour imaging. AB - p-[123I]iodo-L-phenylalanine (IPA) is a recently described radiopharmaceutical which is highly accumulated in gliomas. The present investigation was designed to evaluate the feasibility of single photon emission tomography (SPET) with IPA to image brain tumours under routine clinical conditions. Using a dual- and a triple headed SPET camera, whole-body kinetic and brain SPET, as well as plasma, urinary and dosimetric analysis were determined in four patients with gliomas after intravenous injection of IPA. Results obtained by IPA SPET were retrospectively compared with histopathology, magnetic resonance imaging and positron emission tomography with [18F]fluorodeoxyglucose. Tumour lesions were clearly demonstrated by IPA SPET at 30 min, 1h and 4.5h post-injection, even in patients with low grade gliomas. In patients with glioblastoma, excellent visualization of the tumour was possible even at 7h p.i., indicative of the high retention of the radiopharmaceutical in cerebral gliomas. Analysis of the radioactivity in plasma and urine attested to the high in vivo stability of IPA. Blood clearance was rapid (> 65% after 10 min) and IPA was excreted predominantly by the kidneys, the urinary radioactivity excretion ranging from 27% at 1h to 54% of injected doses at 5h p.i. The average effective dose for adults was estimated to be 0.0152mSv*MBq(-1), leading to an effective dose of 3.8mSv in a typical brain SPET investigation with 250 MBq IPA. This result strongly suggests that IPA is a potentially valuable brain tumour imaging agent for widespread clinical studies with SPET. Its high specific tumour uptake and retention even in low grade gliomas represent a major advantage compared to presently available SPET radiopharmaceuticals. Moreover, the radiation dose estimates indicate that clinical use of IPA will result in acceptable radiation dose levels in humans. PMID- 11891466 TI - Clinical measurement of blood flow in tumours using positron emission tomography: a review. AB - In oncology drug development there is an increasing need for the in vivo physiological measurement of changes in tumour blood flow in response to therapy. Positron Emission Tomography (PET) is being increasingly used in oncology patients to measure blood flow. Here we review the clinical use of PET to measure vascular parameters in man. PMID- 11891468 TI - Effect of the size of regions of interest on the estimation of differential renal function in children with congenital hydronephrosis. AB - The estimation of differential renal function from dynamic renography affects clinical decisions. The estimation requires the kidneys to be delineated with regions of interest. However, in the presence of unilateral hydronephrosis it is not unusual for the affected kidney to be enlarged so that the regions of interest required to delineate the normal and abnormal kidneys are themselves dissimilar in size. The question, which then arises is, will this difference in the sizes of the regions of interest affect the resultant estimation of differential renal function? Eighteen children aged 1 month to 7 years, with prenatal ultrasound diagnosis of unilateral hydronephrosis where the affected kidney was larger than the normal kidney, underwent a total of 57 diuretic renograms using 99mTc-mercaptoacetyltriglycine. The renographic data were retrieved from optical disc and re-analysed. Regions of interest were generated which enclosed each kidney plus a region of interest of equal size to the abnormal large kidney was placed over the normal kidney. The consensus report from the Scientific Committee of Radionuclides in Nephrology described the Rutland-Patlak plot and integral methods for the estimation of differential renal function from dynamic renography. These two methods were used to analyse renal curves with and without background subtraction. Evaluation of the results suggest that technical factors, including the size of the region of interest, may affect differential renal function, and may, in part, explain the reports of the super normal kidney in the literature. PMID- 11891467 TI - The effect of gadolinium-based MRI contrast agents on the biodistribution of (67)Ga. AB - It has been reported that administration of the paramagnetic contrast agent Gd diethylenetriaminepentaacetic acid (Gd-DTPA, gadopentate) prior to 67Ga citrate could lead to poor quality scans, characterized by pronounced bone uptake and a loss of tumour avidity. Suggestions to account for this behaviour included in vivo dissociation or the presence of free DTPA in the formulation. The objective of this study was to assess this potential interference in 67Ga imaging using a mouse model. Commercial gadopentate and gadodiamide contrast agents at doses up to 5mmol*kg(-1) were injected into mature female Balb/c mice 4h before i.v. 67Ga citrate, then the biodistribution was determined at 24h. Gd-DTPA solutions containing excess Gd or DTPA were examined as well. The model was verified by identical studies using inactive Ga(III) or Fe(III) at 0.1mmol*kg(-1). The effects of Gd(III) or the DTPA ligand at this dose were also determined. Administration of Gd-DTPA was found to produce no marked changes in 67Ga biodistribution. Minor changes occurred after 0.1mmol*kg(-1) Gd(III) or the DTPA ligand, but could not account for the scan changes reported above. Inactive Ga(III) or Fe(III), as expected, caused a marked reduction of 67Ga uptake in all tissues except bone, leading to greatly increased total bone:total soft tissue ratios. It is concluded that Gd-DTPA or its constituents do not significantly alter the biodistribution of 67Ga citrate in mice. Extrapolating these findings to the human situation suggests that the previously reported scan changes may have been the result of other undetermined factors. PMID- 11891470 TI - Significant reduction of 125 I-meta-iodobenzylguanidine accumulation directly caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydroxypyridine, a toxic agent for inducing experimental Parkinson's disease. AB - A significant reduction of cardiac 123I-meta-iodobenzylguanidine (MIBG) accumulation has been reported in patients with idiopathic Parkinson's disease. However, it is unclear whether this reduction in cardiac sympathetic nerve is caused primarily or secondarily to the degeneration of sympathetic nerve centres which occurs in Parkinson's disease. Therefore, we examined neuronal 125I-MIBG accumulation in mice hearts of an experimental Parkinson's disease model and in sympathetic cells without any neuronal innervation. Cardiac accumulation of 125I MIBG was determined 4h after intravenous injection of 125I-MIBG in mice pretreated with 1-methyl-4-phenyl-1,2,3,6-tetrahydroxypyridine (MPTP), an inducer of Parkinson's disease. In an in vitro study, uptake of 125I-MIBG was determined in a cultured pheochromocytoma cell line (PC-12), which was pretreated with MPTP. MPTP reduced MIBG accumulation mainly in its neuronal component of mice hearts, suggesting that MPTP impairs cardiac sympathetic nerves to uptake MIBG. Application of MPTP also caused near-complete blockade of 125I-MIBG accumulation in PC-12 cells. In the experimental PD models, it was shown that neuronal accumulation of MIBG was impaired by the direct action of MPTP to the sympathetic cells. These findings support the idea that cardiac sympathetic nerves are primarily impaired in Parkinson's disease despite the presence or absence of systemic autonomic failure. PMID- 11891469 TI - Imaging of abdominal infection using 99m Tc stannous fluoride colloid labelled leukocytes. AB - Radiolabelling of leukocytes using labelled phagocytosed technetium-99m (99mTc) colloidal radiopharmaceuticals has been reported as a method for imaging infection. This in vivo study compares the use of leukocytes labelled using 99mTc stannous fluoride colloid with leukocytes labelled using indium-111 (111In) oxinate. A total of 26 patients (10 male, 16 female; mean age 52 years, range 23 88 years) referred for the investigation of possible infection were studied using both leukocyte labelling methods simultaneously. Images were acquired 4h and 24h after re-injection of the labelled cells. The images were evaluated qualitatively by two nuclear medicine physicians. The results show a high degree of concordance between the techniques: 11 of the 28 images showed a focus of leukocyte accumulation with both techniques at 24h, and 13 out of 28 showed a normal appearance at 24h with both methods. In four cases the results were discordant; the 99mTc stannous fluoride colloid labelled leukocytes gave a false positive appearance at 24h in three patients and a false negative in one. In conclusion, colloid labelling of leukocytes offers a sensitive method for the detection of infective foci coupled with the high resolution imaging offered by 99mTc. It has the advantage over other in vitro labelling methods of being a simpler, non labour-intensive procedure employing whole blood, and its use should be considered by departments that have limited facilities for in vitro leukocyte labelling. PMID- 11891472 TI - Influence of the physiological changes of gastric emptying on the simplified single sample 14 C-urea breath test. AB - The aim of this study was to assess the influence of the physiological changes of gastric emptying on the simplified 14C-urea breath test. Thirty patients performed the test in fasting conditions. Patients were orally administered 0.074 mega Bq of 14C-urea, mixed with 0.0185 mega Bq of 99mTc-S colloids in 25 ml water. A breath sample was taken before and 10 min after intake of the tracers and followed by a 2 min planar anterior scintigraphic image of the abdomen to measure gastric activity. Gastric emptying was estimated by dividing the residual gastric activity at 10 min by the total activity in the abdomen. The procedure was performed twice for each patient after a 24 h interval. The repeatability of both the gastric emptying test and the urea breath test was assessed by the method described by Bland and Altman. The coefficient of repeatability of the urea breath test was 1.18 for a confidence interval of 95%. The coefficient of repeatability of gastric emptying was 27.4. There was no significant correlation (r= 0.08) between the plot of the individual modifications of urea breath test and residual gastric activity in two successive tests. It is concluded that the physiological changes of gastric emptying do not influence the results obtained by the simplified, single-sample 14C-urea breath test. PMID- 11891471 TI - 99m Tc-ciprofloxacin (Infecton) imaging in the diagnosis of knee prosthesis infections. AB - The purpose of this study was to evaluate the usefulness of 99mTc labelled ciprofloxacin imaging in detecting the presence of infection in patients with symptomatic knee prostheses. Among 16 randomly selected patients of whom seven had infection based on clinical and microbiological findings and nine did not, 99mTc-ciprofloxacin images were obtained at 1, 4 and 24h after the injection of the tracer. While there was some diffuse non-specific accumulation of 99mTc ciprofloxacin in large synovial joints and in prosthetic knee joints, the infected knee prostheses were found to show more intensive focal uptake, which also extended outside the synovial cavity. The infection related uptake remained visible in the 24h images, whereas non-specific uptake had a fading tendency at this time point. 99mTc-ciprofloxacin imaging showed diagnostic sensitivity of 86% and a specificity of 78% for correctly classifying the presence of infection. The data indicate that 99mTc-ciprofloxacin imaging may be used in the diagnosis of knee prosthesis infections. Infection-related uptake remains visible in the 24h images and is typically found also outside the synovial cavity, which should be noted in the evaluation of the images. PMID- 11891473 TI - Estimation of left ventricular systolic pressure by the left ventricular volume time curve obtained from electrocardiograph gated 99m Tc-tetrofosmin single photon emission tomography using quantitative gated SPECT. AB - We report the estimation of left ventricular systolic pressure (LVSP) by a left ventricular (LV) volume-time curve obtained from electrocardiogram (ECG) gated 99mTc-tetrofosmin single photon emission computed tomography (SPECT) using quantitative gated SPECT (QGS). LVSP was calculated based on the following parameters: LV volumes, velocity and acceleration of LV contraction, aortic valve area and density of blood. The first three parameters can be derived from ECG gated SPECT. In 16 patients, the LV volume-time curve was obtained from ECG gated SPECT by using QGS. LVSP was estimated by the above-mentioned theory. The values of estimated peak LVSP were compared with those measured from a pressure transducer. There was a correlation between the values of peak LVSP estimated by the LV volume-time curve and those measured by pressure transducer (r=0.69, P<0.01). Using QGS, LVSP and the systolic LV pressure-volume relationship could be estimated by the LV volume-time curve. PMID- 11891474 TI - Use of linearity of the Sokoloff model to improve performance of non-linear search. AB - The three-parameter Sokoloff equation is used to measure the rates of glucose consumption in the brain in vivo. This equation depends linearly on one of its parameters, k1, which is responsible for the glucose transport from plasma to tissue. By equating to zero the first derivative of the minimization function with respect to k1, it is possible to express this parameter as a function of the other two and reduce the non-linear search from three to two dimensions. This approach was examined by the Nelder-Mead simplex method and the Levenberg Marquardt algorithm. In both cases the process of convergence was more robust and required fewer iterations to achieve a given accuracy than the direct three parameter non-linear search. PMID- 11891475 TI - Comparison of 201 Tl- with 67 Ga single photon emission tomography in the diagnosis of head and neck cancer recurrence. AB - The purpose of this study was to evaluate the usefulness of 201Tl single photon emission tomography (SPET) in comparison with 67Ga SPET for distinguishing recurrent tumours in patients previously treated for head and neck cancer. A total of 37 patients with suspicion of recurrent cancer were investigated. SPET images with 201Tl were acquired 10min (early) and 3h (delayed) and SPET images with 67Ga were acquired 72h, after injection. The visual and semiquantitative (T/N ratio) analysis were performed. On visual analysis, results from early 201Tl SPET were the same as those from delayed 201Tl SPET. The sensitivity, specificity and accuracy of the diagnosis of local recurrence using 201Tl SPET were all 100%. The three values using 201Tl SPET for neck lymph node metastases were 73%, 100% and 91%. The corresponding values using 67Ga SPET for local recurrence were 57%, 100% and 89%, respectively, and those using 67Ga SPET for neck lymph node metastases 55%, 100% and 84%, respectively. In the semiquantitative analysis, there was a statistically higher T/N ratio obtained using 201Tl when compared with 67Ga. 201Tl early SPET, especially, has the potential to replace 67Ga SPET in the follow-up of patients with head and neck cancer. PMID- 11891476 TI - Is there a future for adrenal scintigraphy? PMID- 11891477 TI - Efficiency comparison between 99m Tc-tetrofosmin and 99m Tc-sestamibi myocardial perfusion studies. AB - The purpose of this investigation was to compare the efficiency of two different imaging protocols using two different clinically available 99mTc labelled myocardial perfusion tracers. One thousand one hundred and thirty-four imaging studies were performed prospectively, using either 99mTc-tetrofosmin or 99mTc sestamibi, alternating the use of each tracer for a total period of 8 months. 99mTc-tetrofosmin rest studies were performed with injections of 259MBq-370MBq and imaging 30 min later. Exercise studies were performed with injections of 777MBq-1.11GBq and imaging 20 min later. 99mTc-sestamibi studies used doses similar to those in the 99mTc-tetrofosmin studies. Imaging followed a standard procedure, at 60 min after rest injection, and 30 min after exercise. For patients undergoing pharmacological stress testing99mTc-sestamibi was imaged 45 min after injection and 99mTc-tetrofosmin was imaged 30 min after injection. Variables analysed were (1) injection-to-imaging time for the procedure, and (2) the number of repeated scans because of extra cardiac activity. The completion time for the rest study was significantly shorter for 99mTc-tetrofosmin compared to 99mTc-sestamibi (47.7+/-21.7 min vs 74.3+/-25.8 min P<0.0001). Likewise, the total study time was shorter for 99mTc-tetrofosmin compared to 99mTc-sestamibi (90+/-32.7 min vs 124+/-37 min, P<0.0001). More importantly, the number of repeated scans was higher with 99mTc-sestamibi compared to 99mTc-tetrofosmin, 21.4% vs 10%, P=0.001 for rest studies and 16.4% vs 7.9% P=0.001 [corrected] for rest and stress. It was concluded that, using a same day rest/stress protocol, 99mTc-tetrofosmin provided higher patient throughput with fewer repeat scans. These factors may be considered for efficiency improvement in nuclear cardiology laboratories using 99mTc perfusion tracers. PMID- 11891478 TI - 14 C-deoxyglucose imaging overestimates myocardial viability in subacute infarction of rats. AB - Clinical studies using 18F-fluorodeoxyglucose suggest that this tracer may overestimate myocardial viability. This study aimed to elucidate whether 2 deoxyglucose accurately indicates myocardial viability at the early phase of myocardial infarction. Autoradiography with 14C-deoxyglucose was performed in fasting rats whose left coronary artery was occluded for 60 min and then reperfused. 14C-deoxyglucose was injected 30 min after the reperfusion (acute; n=10) or 1 week later (subacute; n=9). Infarction and risk areas were identified by triphenyl tetrazolium chloride or haematoxylin-eosin staining and methylene blue, respectively. Immuno-histochemical staining using anti-glucose transporter 1 and 4 antibodies was performed. At the acute stage, the uptake of deoxyglucose was consistent with the grade of anti-glucose transporter 4 expression. At the subacute stage, the uptake of deoxyglucose in poorly viable myocardium (543.4+/ 343.7%: normalized with the uptake at the right ventricle) as well as in the viable one (335.2+/-149.8%) in the risk area was significantly greater than that in the remote area (116.4+/-94.9%, P<0.01). Anti-glucose transporter 1 was expressed in the poorly viable area where inflammatory cells infiltrated. It is concluded that deoxyglucose uptake by inflammatory cells which express anti glucose transporter 1 causes overestimation of myocardial viability at subacute stage. PMID- 11891479 TI - Quantification of myocardial hypoperfusion with 99m Tc-sestamibi in patients undergoing prolonged coronary artery balloon occlusion. AB - Percutaneous transluminal coronary angioplasty provides an excellent opportunity to investigate the location and quantity of hypoperfusion during sudden complete occlusion of one of the major coronary arteries. Thirty-five patients referred for elective percutaneous transluminal coronary angioplasty were injected intravenously with 99mTc-sestamibi during balloon inflation. To visualize and quantify the hypoperfused region, a map of perfusion was constructed from that occlusion study and from the control study performed on the following day. Patients were divided into groups according to proximal or distal occlusion within each of the three coronary arteries. The region of myocardium supplied by each coronary artery varied in location and extended outside the typical borders for all arteries, but most prominently for the left circumflex coronary artery. The quantities of hypoperfusion varied within each artery group, but the average hypoperfusion was greater for the left anterior descending coronary artery than for either the right coronary artery or the left circumflex coronary artery. It is concluded that the quantities of hypoperfusion were highly variable within each artery group. Occlusion of the left anterior descending coronary artery was associated with the largest ischaemic region. The area of hypoperfusion extended outside the typical borders, most prominently for the left circumflex coronary artery. PMID- 11891480 TI - 131 I-cG250 monoclonal antibody immunoscintigraphy versus [18 F]FDG-PET imaging in patients with metastatic renal cell carcinoma: a comparative study. AB - The aims of this study were to establish the percentage of metastatic renal cell carcinoma (RCC) lesions detected by radioimmunoscintigraphy (RIS) with the chimeric monoclonal antibody 131I-cG250 versus positron emission tomography (PET) with 18F-labelled deoxyglucose ([18F]FDG), and to evaluate the use of these radionuclide imaging modalities compared with routinely used imaging techniques. Twenty patients with metastatic RCC disease were examined with [18F]FDG-PET and 131I-cG250 RIS within 1 week. Total body gamma camera images were obtained up to 120h after injection of 232MBq 131I-cG250. Total body PET scanning was performed 45-60 min after intravenous injection of 370MBq [18F]FDG. Nuclear medicine techniques were compared to routine imaging procedures. Routine imaging modalities revealed a total of 79 metastases. [18F]FDG-PET and 131I-cG250 RIS detected 33 previously unknown metastases, of which 32 were [18F]FDG positive and seven were 131I-cG250 positive. Of the 112 tumour lesions that were documented, [18F]FDG-PET detected 69% (77 out of 112), whereas 131I-cG250 RIS detected only 30% (34 out of 112). In conclusion, [18F]FDG-PET is superior to 131I-cG250 RIS in detecting metastases in patients with metastatic RCC, and therefore seems a promising tool for (re)staging patients with RCC. The usefulness of RIS with a diagnostic dose of 131I-cG250 seems to be restricted to selecting patients for radioimmunotherapy with 131I-cG250. PMID- 11891481 TI - Synthesis of 188 Re-labelled long chain alkyl diaminedithiol for therapy of liver cancer. AB - Radioisotope-labelled lipiodol has been used in the therapy of liver cancer. Recently a lipiodol solution of 188Re-labelled diaminedithiol (DD) has been reported to show a high uptake in the liver cancer. We synthesized long-chain alkyl DD derivatives to improve their uptake and retention in tissue. As the length of the alkyl chain increased, tissue uptake and retention also increased due to hydrophobic interaction with lipiodol. Among the synthesized compounds, the lipiodol solution of 188Re-HDD, the DD derivative with the longest side chain (C16), is a promising agent for therapy of liver cancer. PMID- 11891482 TI - Preparation and evaluation of 90 Y skin patches for therapy of superficial tumours in mice. AB - 90Y, a beta emitting radionuclide, was immobilized in a bandage patch for possible application for the therapy of superficial maladies such as tumours and skin cancers. The aim was to prepare a radiation source that could deliver uniform dose within a short duration and avoid the inconveniences faced with the gamma sources used in teletherapy and brachytherapy. 90Y-ferric hydroxide macroaggregates were prepared, filtered and immobilized between two layers of gauze. When placed in saline, no radioactivity leached for 3 days, proving its safety for external application. Fibrosarcoma was induced in mice for checking the efficacy of 90Y patches. 90Y patches of various activities were prepared and applied on the tumours. The effect of time gap between inception and treatment of tumour, dose delivered and multiple application in fractionated doses was studied. In all the cases, tumour growth in the treated animals was considerably reduced in comparison with controls. It was concluded from the experiments that treatment should be started at the earliest stage possible, i.e. when the tumour is palpable. Delivery of dose from radioactive patches of approximately 90-100 MBq each, thrice at weekly intervals proved to be more effective for regression of tumour growth. PMID- 11891484 TI - Effects of age and lateralization on lymphoscintigraphic interpretation. AB - The present study was undertaken to find out if ageing and lateralization might influence the results of lymphoscintigraphic investigations. Axillary lymphoscintigrams obtained in 756 women after subcutaneous intercostal (IC) injection of 99mTc-labelled colloids at the level of the chest wall were reviewed and analysed according to age (<41 years, 41-50 years, 51-60 years, 61-70 years, 71-80 years, >80 years) and the side injected (right or left). No axillary nodes were visualized (IC-) in 34% of the population, and IC- cases were somewhat, but not significantly (0.1080 years, four out of seven), and the absence of drainage was more common in patients over 50 years old (overall, 38.2%; right, 36%; left, 40.2%) than in younger cases (overall, 27.9%; right, 24.1%; left, 30.3%). From a statistical point of view, the differences between these two age discriminated populations were significant both when considering the series of injections as a whole (0.01>2P>0.001) and injections in the right side only (0.01<2P<0.02). In conclusion, ageing and lateralization influence lymphoscintigraphic investigations and have to be taken into account when analysing results. PMID- 11891483 TI - Bone marrow scintigraphy with 99m Tc labelled monoclonal anti-NCA 90 Fab' fragment: a feasibility study and comparison of bone marrow uptake with 99m Tc labelled monoclonal anti-NCA 95 antigranulocyte antibody. AB - The aim of this retrospective study was to evaluate the usefulness of 99mTc labelled monoclonal anti-NCA 90 antigranulocyte antibody Fab' fragment (MN3 Fab') as a bone marrow imaging agent. One hundred and ten planar scans (88 patients) of the lumbar and sacroiliac regions as well as whole-body scans were performed after 1, 5 and 24 h. All the scans were evaluated visually and bone marrow uptake was determined semiquantitatively as count density ratio from sacroiliac-minus background to background area. Results were compared to 50 age-matched patients with normal bone marrow scans obtained with the intact 99mTc labelled monoclonal anti-NCA 95 antigranulocyte antibody (BW 250/183) in a previous study. Seventy three patients showed a physiological activity distribution in the central bone marrow. Ten patients showed a bone marrow extension, while in two patients central bone marrow depression was observed. Evaluation of the ribs, lower thoracic and upper lumbar spine was hampered by soft-tissue activity. Bone marrow uptake was 1.36+/-0.56 after 1 h, decreased thereafter and was significantly lower than that of BW 250/183 (P < 0.001). In conclusion, MN3 Fab' cannot be recommended for bone marrow scintigraphy, because relevant parts of the haemopoietically active bone marrow are not accessible to visual evaluation. A significant role of the semiquantitative evaluation of MN3 Fab' bone marrow uptake in patients with potential marrow depression seems unlikely. PMID- 11891485 TI - Radiation protection issues of treating hyperthyroidism with 131 I in patients on haemodialysis. AB - We report on the cases of two patients referred for 131I treatment of hyperthyroidism who were dependent on haemodialysis. Following 131I administration, all disposable lines and filters from dialysis were collected and measured for 131I radioactivity. The amount of 131I retained by the filters at the end of each successive dialysis session was found to decay with effective half-lives of 6.6+/-0.2 and 6.3+/-0.2 days. Dose rate measurements at 1m from the patients were recorded to find the effective half-life of the radioiodine clearance, which were found to be 6.9 and 7.1 days. From measured dose rates taken at 30 cm, the radiation hazard to ward staff involved in patient management was shown to be negligible. PMID- 11891486 TI - Renal uptake rate measurement of 99m Tc-dimercaptosuccinic acid using spectral analysis. AB - We developed a new method for measuring the rate of renal uptake of 99mTc dimercaptosuccinic acid (DMSA) using spectral analysis. The renal uptake rate (per minute) of DMSA (K) was calculated by averaging the tissue impulse response function values obtained by spectral analysis between 10 min and 15 min. The K values obtained by spectral analysis correlated well with the renal uptake rates (%) measured 2h after DMSA administration (r=0.921 with background correction; r=0.924 without background correction). There was a good agreement between the K values obtained by spectral analysis using the kidney time-activity curves with (x) and without (y) background correction (r=0.993, y=1.089x+0.004), suggesting that our method requires no background correction. There was excellent correlation between the K values obtained by spectral analysis using the kidney time-activity curves with (y) and without (x) kidney depth correction (r=0.992, y=1.721x+0.000 with background correction; r=0.990, y=1.720x+0.000 without background correction), suggesting that our method requires no kidney depth correction. These results indicate that spectral analysis is appropriate and useful for the quantification of renal uptake rate of DMSA. We believe that this method will facilitate even more widespread utilization of the quantitative assessment of DMSA uptake by planar scintigraphy, since it needs only 10-15 min for imaging, and background and kidney depth correction and blood sampling are not required. PMID- 11891487 TI - Determining the influence of biocompatible oxygenators during cardiopulmonary bypass by 99m Tc-DTPA aerosol assessment of pulmonary permeability. AB - Changes in pulmonary permeability provide a partial measure of the clinical impact of biocompatible oxygenator use during cardiopulmonary bypass surgery. Previous research has shown that the clearance rate of 99mTc-labelled diethylene triamine penta-acetic acid (99mTc-DTPA) aerosol from the lungs is increased following cardiopulmonary bypass, resulting from an increase in pulmonary permeability. The aerosol clearance rate has been shown to return to normal after a period of 7 days. A blind trial was set up to assess the clinical impact of a biocompatible, Trillium-coated oxygenator compared with a standard oxygenator. In a group of 25 patients 99mTc-DTPA aerosol studies were carried out prior to cardiopulmonary bypass surgery for mitral valve surgery. Repeat studies were undertaken 3-4 h and 24-28 h after surgery. Analysis of the rates of pulmonary clearance reproduced the trends seen in earlier research. There was however no statistically significant difference in the variation of serial clearance times between the groups of patients undergoing surgery using the Trillium-coated oxygenators and those using the standard oxygenators. PMID- 11891489 TI - The temperament profiles of school-age children. AB - Maternal reports of child temperament were used to develop temperament profiles of school-age children. The subjects were 883 children who were between 4 and 12 years of age. The children's families varied substantially in their socioeconomic status and race/ethnicity. To develop the profiles, the dimensions derived from the School-Age Temperament Inventory were subjected to a second order principal factor analysis with varimax rotation. Pearson chi-squares were used to determine whether sociodemographic variables were proportionally represented among the profiles. Forty-two percent of the children were classified into four temperament profiles. High maintenance and cautious/slow to warm up were deemed as challenging temperaments. Industrious and social/eager to try were mirror images of those profiles and were labeled easy. Some children were both types of challenging or easy profiles. The generalizability of the profiles in relation to the sociodemographic variables of gender, age, race/ethnicity, and socioeconomic status was also examined. Challenging temperament profiles were disproportionately represented by boys, Hispanic children, and those from lower socioeconomic families. Girls were over represented in the group that included both types of easy temperaments. Social/eager to try children were more often from higher rather than lower socioeconomic status families. Clinical applications and research implications for the profiles are discussed. The profiles can be used as exemplars that parents can use to recognize their child's temperament. Further research is needed to explore whether different developmental outcomes are associated with the profiles. PMID- 11891490 TI - Transforming temperament profile statistics into puppets and other visual media. AB - This article describes the evolution of visual media that was developed to enhance a clinical intervention called INSIGHTS into Children's Temperament. First, the theoretical and research studies that supported the intervention are presented. Then the iterative steps used to transform the statistical data that identified common school-age children's temperament profiles into puppets and other visual media are enumerated. Finally, the implications of conducting a theory-based intervention within a school environment are discussed. PMID- 11891491 TI - A natural history study of adolescents and young adults with sickle cell disease as they transfer to adult care: a need for case management services. AB - Life expectancy for adolescents with SCD now extends well into adulthood. As a result, adolescents transfer to adult care. Little empirical evidence exists to show how transfer occurs and how well the current practices now work. The aim of this study was to obtain a database on the experience of adolescents/young adults with SCD that transfer to adult care. We assessed their treatment compliance, independence, and whether they receive uninterrupted care. Data were obtained through patient and provider interviews and patient record reviews. Results indicate patients leave pediatric care without adequate transfer preparation and readiness to transfer is not the major consideration in the decision to transfer, follow-up often ceases once the patients leave pediatric care, and patients who maintain follow-up appointments are more efficient in managing their illness (self-efficacy). The model for a structured transitioning process is provided with recommendations for nurse case managers to maintain follow-up. PMID- 11891492 TI - Psychosocial factors associated with levels of metabolic control in youth with type 1 diabetes. AB - This study sought to determine the relationship between Youth Self Report (YSR) scores for behavior problems, YSR scores for social competence, and metabolic control in children and adolescents with Type 1 diabetes. Using a cross sectional design, 234 individuals between 11 and 18 years old were given the YSR at regular clinic appointments; glycosylated hemoglobin (GHb) was also measured. More than 50% of subjects showed GHb levels above 9%; the normal GHb level is 4.2% to 5.8%. Individuals reporting a greater number of behavior problems, though not in the pathological range on the aggression, delinquent behaviors, and attention problems subscales of the YSR, were more than twice as likely to have GHb levels above 9%. Using logistic regression the externalizing scale (aggression and delinquent behaviors combined) predicted elevated GHb at 2.41, p =.003. Youth in this study were middle-class and were receiving subspecialty care. Yet, over half of them had GHb levels above the recommended 9%. The psychological health of youth should be monitored at regular intervals. Longitudinal studies are needed to determine whether aggression, delinquent behaviors, and attention problems in children and adolescents with Type 1 diabetes result later in depression and elevated levels of GHb in these same individuals or whether these elevations are transient. Interviews could be supplemented with instruments such as the YSR and care given for those with a higher number of self-reported problems. PMID- 11891493 TI - Multisite research: a systems approach. AB - Multisite research is becoming increasingly common because of the need for an adequate sample size and for generalizability of results beyond a single facility. Collaboration in a research project poses unique challenges due to the number of persons and facilities involved. The use of a systems approach to structure the research process in a study of pain in children with leukemia is described, using the principles to structure, conduct, and conclude the multisite project. PMID- 11891494 TI - Advocacy and leadership when parental rights and child welfare collide: the role of the advanced practice nurse. AB - This article describes the experience of an advanced practice nurse in a challenging clinical situation. A mother with mental illness and mental retardation seeks to retain parental rights and care for her newborn with cystic fibrosis. The nurse provides leadership to the hospital team and serves as an advocate throughout legal proceedings. A systematic, nonjudgmental, and empathic approach to gathering information, working with the family, welfare, and legal representatives is described. Enacting a complex and court-mandated homecare education regimen to the disabled mother is discussed. Preparation to testify in a termination of parental rights proceeding is outlined and a summary description of the testimony provided. PMID- 11891495 TI - The silent epidemic: lead poisoning. PMID- 11891496 TI - Children and terror. PMID- 11891497 TI - Dante's orphans. PMID- 11891500 TI - Introduction. PMID- 11891498 TI - The private adolescent: privacy needs of adolescents in hospitals. AB - This report is based on an interpretative study that explored the meaning of space to adolescents in the hospital environment. Through designing their own adolescent ward and discussing their designs in an interview, participants articulated their spatial needs in the ward environment. This paper addresses the private space issues of the adolescent patient in the ward environment. Issues that are discussed include the use of the telephone, the bathroom, and the bedroom, and additional facilities needed to enhance privacy in the ward. PMID- 11891501 TI - Monogenic forms of human hypertension. AB - Monogenic or Mendelian forms of hypertension have ushered in a revolution in our knowledge. If we add information on syndromes involving low blood pressure, this knowledge base is doubled. Glucocorticoid-remediable aldosteronism, apparent mineralocorticoid excess, and mutations in the mineralocorticoid receptor gene have given us brilliant insights into mineralocorticoid-induced hypertension. The latter discovery has elucidated how mutations may modify the receptor sufficiently to allow erstwhile antagonists to have an agonistic action. The epithelial sodium channel (ENaC) has been elucidated. Gain-of-function mutations in the beta and gamma subunits of ENaC cause Liddle's syndrome. Loss-of-function mutations in all 3 subunits of ENaC cause hypotension (pseudohypoaldosteronism type I). Thus, all 3 subunits can be mutated, causing either hyper- or hypotension. Three loci have been described for Gordon's syndrome, pseudohypoaldosteronism type II; 2 members of the WNK (with no ly sine K) serine threonine kinase family have recently been found to be responsible. Autosomal dominant hypertension with brachydactyly features normal sodium and renin angiotensin-aldosterone responses. The gene has been mapped to chromosome 12p. The condition is interesting because it may represent a novel neural form of hypertension. The elucidation of Mendelian blood pressure-regulatory disorders has been a resounding success. PMID- 11891502 TI - How many pathways to pheochromocytoma? AB - Pheochromocytomas, like several other tumors, may be either sporadic or the manifestation of a familial cancer syndrome. Recently, major advances have occurred in both the understanding of diverse molecular mechanisms leading to pheochromocytoma and the diagnostic modalities available for detection of the disease. Familial pheochromocytoma may be a manifestation of multiple endocrine neoplasia type 2 (MEN-2), von Hippel-Lindau (VHL), or neurofibromatosis-1 (NF 1) disease. Tumor-suppressor genes responsible for the familial occurrence of extra adrenal pheochromocytoma, called paraganglioma, have been identified. This wealth of genetic information, coupled with the availability of sensitive and specific biochemical tests as well as imaging studies, allows for genetic screening and early diagnosis of pheochromocytoma. In addition, genetic screening of relatives at risk is now feasible. In this article, we review recent clinical and molecular advances in our understanding of pheochromocytoma. PMID- 11891503 TI - Genetics of obesity and obesity-related hypertension. AB - Obesity results from an imbalance between caloric intake and energy expenditure. Twin, adoption, and family studies have not only shown that genetic factors play an important role in the pathogenesis of obesity, but also contribute to several comorbidities including hypertension and type 2 diabetes. In recent years, several single-gene defects responsible for obesity in rodents and, in rare cases, of human obesity have been identified. Besides leptin as the most notable example, numerous other proteins and neuropeptides have recently been found that participate in a complex network to regulate food intake and energy expenditure. Interestingly, some of these molecules may also play a role in the development of obesity-related hypertension. The ongoing search for relevant genetic variants should result in a better understanding of energy metabolism and hopefully clarify molecular mechanisms underlying the association between obesity and related comorbidities. This knowledge should help develop new strategies for the treatment of obesity and associated risk factors for hypertension and related cardiovascular disorders. PMID- 11891504 TI - Statistical gene mapping of traits in humans--hypertension as a complex trait: is it amenable to genetic analysis? AB - Gene mapping refers to the localization of disease genes on the human gene map. The currently used mathematical methods for gene mapping are outlined and compared. The example of Gilles de la Tourette syndrome is discussed in detail. PMID- 11891505 TI - Hypertension as a complex genetic trait. AB - Pickering first showed that blood pressure distribution is unimodal and that the diagnosis of essential (primary) hypertension is an arbitrary quantitative trait. His family studies suggested that not 1, but many, perhaps 30 or more, gene variations are responsible for raising blood pressure. Two approaches have been used to address the genetics of essential hypertension. Candidate genes have been selected by virtue of their physiologic function, and case-control association studies have been performed. Numerous candidates have been evaluated, genes of the renin-angiotensin-aldosterone system, genes coding for adrenergic receptors, genes coding for proteins regulating endothelial function, and genes involved in signaling have been studied. True to Pickering's prediction, each gene tested thus far seems to exert, at best, a small effect and contradictory studies are common. Linkage studies have been performed to map the loci of genes regulating blood pressure or inducing hypertension. Studies of dizygotic twins and their parents have permitted an identity-by-decent linkage analysis. Studies of affected sibling pairs involve subjects whose parents are generally already dead. Identity-by-state analysis requires a far greater number of pairs for results. Nevertheless, some large linkage studies have now been performed including subjects from Framingham. Log odds (LOD) scores are far removed from actually cloning genes responsible for hypertension. Single nucleotide polymorphisms (SNPs) permit linkage dysequilibrium mapping, which may enable cloning new genes. Thus far, very few new genes have been cloned for any complex genetic disease. The task is daunting but not impossible. PMID- 11891507 TI - Congenic rat strains are important tools for the genetic dissection of essential hypertension. AB - The identification of mutated genes and the characterization of disease pathways in monogenetic forms of human arterial hypertension has been very successful during the past decade, although only little progress has been made with regard to the genetic analysis of essential hypertension. Inbred rat strains that display hypertension as an inherited trait represent an attractive substitute for the dissection of the polygenetic basis of human essential hypertension. The power of studying these inbred models with natural allelic variation is that each strain can be thought of as a different subtype of essential hypertension. Thus, to study the genetics of hypertension using a potpourri of inbred hypertensive rat strains may help to identify a set of genes that can give rise to hypertension in the heterogeneous clinical situation of essential hypertension. Indeed, multiple blood pressure quantitative trait loci (QTL) have been identified in the rat genome and a few of those have been implicated in the human disease by homology mapping already. Here, we discuss the role of congenic breeding strategies to further dissect the genetics of hypertension in the rat. Based on recent accomplishments it can be anticipated that by using congenic strains, the genetic sifting of the collection of blood pressure QTLs identified so far will lead to the identification of additional and new candidate genes. This will open the possibility for the development of new diagnostic and therapeutic means contributing to a better control of human essential hypertension. PMID- 11891506 TI - Genetic and gender determinants of cerebrovascular disease. AB - This article reviews genetic studies of stroke, bearing insight into human evidence by using studies on twins as well as on monogenic forms of stroke. Focus will be given to inbred animal models that have been used to improve genetic homogeneity and to control environmental influences. Such animal studies have lead to the identification of quantitative trait loci harboring putative genes for stroke susceptibility and sensitivity. The major strategies for gene identification are discussed. Experimental animal models of stroke have also revealed a sexual dimorphism in stroke susceptibility and sensitivity and this article reviews 2 possible candidates, namely, the Y chromosome and estrogens. PMID- 11891508 TI - Genetics of Cd36 and the hypertension metabolic syndrome. AB - Although genetic mapping of quantitative trait loci (QTL) for complex traits related to hypertension is relatively straightforward, the identification of QTL at the molecular level has proven far more difficult. By combining techniques of gene mapping and gene expression profiling with studies in congenic and transgenic strains, a specific molecular defect in the Cd36 fatty acid transporter has been identified that contributes to the pathogenesis of 2 complex traits in the spontaneously hypertensive rat (SHR), namely, insulin resistance and disordered fatty acid metabolism. After mapping QTL linked to insulin resistance and dyslipidemia to the telomeric region of SHR chromosome 4, gene expression studies were used to identify candidate genes within the target chromosome segment that were differentially expressed in white adipose tissue between SHR congenic strains. This led to the identification of a major mutation in the SHR gene encoding Cd36, a fatty acid transporter involved in the transmembrane transport of long-chain fatty acids. The role for mutant Cd36 in the pathogenesis of insulin resistance and dyslipidemia was confirmed by rescuing the abnormal metabolic phenotypes through transgenic expression of wild-type Cd36 on the SHR background. These findings show that a primary defect in fatty acid transport can promote disordered carbohydrate metabolism in the SHR and show the power of advanced genome technologies for identifying QTL at the molecular level. PMID- 11891509 TI - Use of transgenic and knockout strategies in mice. AB - Over 2 decades ago mouse models were generated with an exogenous gene integrated into its genome to create the first transgenic mice. Since that time, new methods have been developed and old methods improved, allowing investigators more flexibility to answer important questions about physiology and gene function. Transgenic and knockout technology have been particularly useful in the kidney as various transgenic mouse models have been successfully generated to gain a better understanding of renal physiology at the gene level. Now with the sequencing of mammalian genomes at the forefront of science, the need for transgenic technology to understand gene function in the context of the whole animal will become increasingly more important. Therefore, this article focuses on some of the strategies that can be used when generating transgenic mouse models. PMID- 11891510 TI - Genetics of programmed cell death and proliferation. AB - Vascular remodeling and hypertrophy-hyperplasia of the heart and kidney are the hallmarks of hypertensions, being involved in the long-term maintenance of elevated blood pressure and the development of cardiovascular and renal hypertension. Both augmented proliferation and unscheduled programmed cell death (apoptosis) contribute to this phenomenon. In the present article, we summarize the results of these studies with an emphasis on the relative impact of genetic determinants and the environment. PMID- 11891511 TI - Adapting renal and cardiovascular physiology to the genetically hypertensive mouse. AB - Gene targeting techniques are a revolutionary tool in determining gene function. The mouse is the only mammal in which these methods can be performed. For morphologic and histologic characterization, a broad armamentarium is available for the mouse. However, for functional studies, the 20- to 25-g mouse provides unique challenges. We have successfully established tail-cuff, intra-arterial, and telemetric blood pressure and heart rate measurements for the mouse. We have also succeeded in measuring renal blood flow, cortical and medullary blood flow, and glomerular filtration rate under various conditions. We have been able to make sophisticated assessments of myocardial performance by using direct intraventricular determinations of pressure and volume. Other investigators have established micropuncture and microperfusion techniques for the mouse. Imaging techniques for the mouse including ultrasound and magnetic resonance imaging are currently feasible. Thus, we will be able to characterize the physiology of any hypertensive mouse model that may become available. PMID- 11891513 TI - Update on pneumococcal infections of the respiratory tract. AB - Respiratory illnesses are the leading reason for seeking medical care here in the United States. Streptococcus pneumoniae is the most common bacterial pathogen causing acute otitis media (AOM), sinusitis, and community-acquired pneumonia in both the pediatric and adult populations. The continued development of antibiotic resistance to an increasing number of different antibiotic classes by this organism has made the treatment of some of these infections more difficult. Recently, a heptavalent pneumococcal conjugate vaccine was approved for infants and toddlers, beginning at 2 months of age. Widespread implementation of this vaccine in the childhood population may have a significant impact on the amount of systemic disease seen with this organism. PMID- 11891514 TI - Mycoplasma and Chlamydia pneumonia in pediatrics. AB - Mycoplasma pneumoniae and Chlamydia pneumoniae are common respiratory pathogens in children 5 years of age and older. Although distinctly different in structure, these organisms share similar epidemiologic and clinical characteristics in human infection and disease. Pneumonia caused by these organisms usually occurs after infection of the upper respiratory tract, but may occur in the absence of antecedent upper respiratory infection. Diagnosis of infection with C. pneumoniae and M. pneumoniae is most often based on clinical findings alone, though definitive diagnosis of infection with either organism may be confirmed through serologic methods, culture, and nucleic acid-detection methods such as polymerase chain reaction. Macrolide antibiotics are highly effective in the treatment of infected children, leading to rapid clinical resolution and excellent long-term out-come in the majority of patients. PMID- 11891515 TI - Respiratory syncytial virus infections in children. AB - Respiratory syncytial virus (RSV) is the leading cause of lower respiratory disease in young children in both developing and developed countries. By age 2, nearly all children have been infected by RSV.The clinical manifestations range from mild upper respiratory symptoms to bronchiolitis and pneumonia. First infections are nearly always symptomatic and frequently cause lower respiratory tract disease, whereas subsequent infections are generally milder. Although children with underlying conditions such as prematurity, chronic lung disease, congenital heart disease, and immuno-suppression are at high risk for severe disease, many children without underlying conditions require hospitalization. Treatment is supportive. Immunoprophylaxis with palivizumab or RSV immune globulin may benefit children born prematurely, especially those with bronchopulmonary dysplasia. To date, the development of an effective vaccine has been unsuccessful. PMID- 11891516 TI - Pneumonia in the immunocompromised pediatric cancer patient. AB - Infectious complications including pneumonia remain a major obstacle to survival in children with cancer. Pulmonary infections usually arise from aspiration of pathogens from the upper airways or from hematogenous spread. Pathogens include bacteria, Pneumocystis carinii, viruses, and fungi. In this article, we review in detail the pathogenic basis, evaluation, and management of pneumonia in the immunocompromised pediatric cancer patient. PMID- 11891517 TI - Pulmonary infections in children with HIV infection. AB - The epidemic of pediatric acquired immunodeficiency syndrome (AIDS) in the United States, which peaked during the mid-1980s and early 1990s, was characterized by a variety of opportunistic infections in children infected with human immunodeficiency virus (HIV), often as the presenting illness of their HIV infection. Pneumocystis carinii pneumonia (PCP) during infancy was responsible for significant morbidity and mortality, followed by many other opportunistic infections, including recurrent, serious bacterial infections; disseminated cytomegalovirus infection; and disseminated Mycobacterium avium complex (MAC) infection. Many of these infections involve the lower respiratory tract either as a primary site of infection or as one of the sites involved in disseminated disease. Since the mid- to late 1990s, the pediatric HIV epidemic in the United States has witnessed a dramatic decrease in the frequency of most opportunistic infections and other severe manifestations of HIV infection in children, primarily because of lower rates of mother-to-child HIV transmission, development and implementation of guidelines for PCP prophylaxis, and availability of highly active antiretroviral therapy. Far fewer children are at risk for clinical progression of HIV disease and for opportunistic infections. Despite these successful trends, pulmonary opportunistic infections and pulmonary disease remain common clinical manifestations of pediatric HIV disease. PMID- 11891518 TI - Pulmonary infections in patients with cystic fibrosis. AB - Cystic fibrosis (CF) is an autosomal-recessive disorder and affects about 60,000 people worldwide. The CF gene, the cystic fibrosis transmembrane conductance regulator (CFTR), was found in 1989 and over 800 mutations have been sequenced. Although our understanding of the pathophysiology of CF has increased, pulmonary infections remain the major cause of morbidity and mortality. During the first decade of life, Staphylococcus aureus and nontypable Haemophilus influenza are most common, but Pseudomonas aeruginosa may be the first pathogen isolated in infants. By 18 years of age, 80% of patients harbor P. aeruginosa and 3.5% harbor Burkholderia cepacia. Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and nontuberculous mycobacteria may be newly emerging CF pathogens. The traditional approach to managing patients with CF is to treat acute pulmonary exacerbations with intravenous antimicrobial agents. However, prophylactic strategies to prevent initial infection or to delay chronic infection with P. aeruginosa or chronic maintenance therapy to slow deterioration of lung function may also improve clinical status. Recognition of the role of inflammation, even early in life, in the absence of clinical symptoms, has led to treatment with anti-inflammatory agents. Novel strategies to disrupt biofilm formation, stimulate chloride conductance, and replace abnormal CFTR are currently being studied. PMID- 11891519 TI - Old and new antibiotics for pediatric pneumonia. AB - A variety of antibiotics, both parenteral and oral, are available to the clinician caring for a child with pneumonia. Although viral pathogens are the common etiologic agents causing community-acquired pneumonia, significant morbidity and mortality exists from disease caused by bacteria and atypical pneumonia agents. Treatment of nosocomial bacterial pneumonia has become particularly difficult with ever-increasing resistance documented in hospital acquired organisms. This article discusses antibiotic therapy based on clinical presentation and based on identified pathogens, with a discussion of mechanisms of antibiotic resistance and the newer agents that have been designed to meet this continually evolving challenge. PMID- 11891520 TI - Emergency department management of acute respiratory infections. AB - Respiratory tract infections are one of the most common problems prompting visits to the emergency department. Although many are the result of self-limited viral illnesses, these infections may result in substantial morbidity and, rarely, mortality. Prompt recognition and appropriate treatment can reduce morbidity and largely prevent mortality. Careful selection of antimicrobial agents is essential to maximize benefit and prevent overuse, which contributes to the emergence of antibiotic resistance. PMID- 11891521 TI - Chronic and recurrent pneumonias in children. AB - The diagnosis and management of chronic and recurrent pneumonia in children may present a significant challenge for the primary care physician. Successful management depends on a careful evaluation of each episode, with a complete review of all available chest radiographs. Timing, location, and prodromes to recurrence can all provide important clues to the etiology of infection. Infiltrates that recur in a single lobe or segment of the lung may be caused by local airway obstruction, or by anatomic abnormalities of the lung itself. Pneumonias that occur in varied locations, or affect more than one lobe, suggest the presence of a more generalized abnormality, such as swallow dysfunction or aspiration, immunodeficiency or asthma. The pattern, frequency of recurrence, and severity of the infections can guide the practitioner in choosing the diagnostic studies most likely to identify an underlying etiology for recurrent episodes of pneumonia. With diligence and patience, most children with recurrent lower respiratory disease can be treated effectively. PMID- 11891523 TI - Microdissection based high resolution multicolor banding for all 24 human chromosomes. AB - The multicolor-banding (MCB) approach allows the differentiation of chromosome region specific areas at the band and sub-band level and is based on region specific microdissection libraries producing changing fluorescence intensity ratios along the chromosomes. The latter are used to assign different pseudocolors to specific chromosomal regions. We present the complete set of 138 region-specific microdissection libraries for the entire human genome and the resulting MCB patterns for all human chromosomes at the 450-550 band level. In the present work, the creation and handling of the microdissection libraries is detailed for the first time. Additionally, the unique possibilities of the MCB technique to adjust the pseudocolor bands according to the necessities of the studied case is presented in exemplarity. In conclusion, the MCB-technique is a high resolution alternative to other FISH based chromosome banding approaches and suited to clarify, which changes appeared in complex chromosomal rearrangements. PMID- 11891522 TI - Diagnostic case study: Fever and patchy infiltrates: pulmonary septic emboli. PMID- 11891524 TI - Analysis of microdissected prostate tissue with ProteinChip arrays--a way to new insights into carcinogenesis and to diagnostic tools. AB - Prostate carcinomas are one of the most common malignancies in western societies. The pathogenesis of this tumor is still poorly understood. These tumors present with two characteristic features: epithelial-mesenchymal interactions, which play a pivotal role for tumor development and most of clinically manifest cancers arise in prostate proper compared to a minority of tumors which develop in the transitional zone. Deciphering the epithelial-mesenchymal cross talk and identification of molecular pecularities of the sub-populations of cells in different zones can therefore help understanding carcinogenesis and development of new, non-invasive tools for the diagnosis and prognosis of prostate carcinomas which has remained a challenge until today. A ProteinChip array technology (SELDI = surface enhanced laser desorption ionization) has been developed recently by Ciphergen Biosystems enabling analysis and profiling of complex protein mixtures from a few cells. This study describes the analysis of approximately 500-1000 freshly obtained prostate cells by SELDI-TOF-MS (surface enhanced laser desorption ionization time-of-flight mass spectrometry). Pure cell populations of stroma, epithelium and tumor cells were selected by laser assisted microdissection. Multiple specific protein patterns were reproducibly detected in the range from 1.5 to 30 kDa in 28 sub-populations of 4 tumorous prostates and 1 control. A specific 4.3 kDa peak was increased in the prostate tumor stroma compared to normal prostate proper and transitional zone stroma and increased in prostate tumor glands compared to normal prostate proper and transitional zone glands. Coupling laser assisted microdissection with SELDI provides tremendous opportunities to identify cell and tumor specific proteins to understand molecular events underlying prostate carcinoma development. It underlines the vast potential of this technology to better understand pathogenesis and identify potential candidates for new specific biomarkers in general which could help to screen for and distinguish disease entities, i.e. between clinically significant and insignificant carcinomas of the prostate. PMID- 11891525 TI - The role of apoptotic resistance in irradiated adult articular chondrocytes. AB - There have so far been no studies on the apoptosis of adult articular chondrocytes after X-ray irradiation. The purpose of this study was to assess the apoptotic resistance of articular chondrocytes in X-ray radiation, in order to examine the possibility of irradiated allogenic chondrocyte implantation. Adult human chondrocytes of the non-degenerated cartilage group without X-ray irradiation did not show positive cells of Annexin V and PI staining in a 48 h culture. The Annexin V positive chondrocytes did not increase in a radiation dose dependent manner, and the PI positive cells were slightly increased at 30 Gy irradiation. In the degenerated cartilage group, the PI positive chondrocytes without irradiation were present, and both the Annexin V and PI positive chondrocytes increased in a radiation dose dependent manner. The Annexin V and PI positive staining of chondrocytes in the non-degenerated cartilage group was less than that of the degenerated cartilage group in the same dose of X-ray irradiation exposure. Loss of the mitochondrial membrane potential, revealed in an early stage of apoptosis, did not show in the irradiated chondrocytes of the non-degenerated cartilage, but were demonstrated in those of the degenerated cartilage. These results demonstrated that the non-degenerated chondrocytes of X ray irradiation were highly resistant for apoptosis, and this knowledge could be applied to allogenic chondrocytes implantation. PMID- 11891527 TI - Evidence for interphase DNA decondensation transverse to the chromosome axis: a multicolor banding analysis. AB - In the present study a multicolor banding (MCB) analysis was performed to address the up to now unrequited question in which direction with respect to the axis chromosomes decondense in interphase. It could be demonstrated, that i) MCB produces a similar banding pattern in interphase as in metaphase; ii) that no complete decondensation and dispersion appears, which is in concordance with the concept of chromosome territories; and iii) chromosome decondensation happens square to chromosome axis. The presented data are important for a better understanding of nuclear architecture, however, further studies are required. PMID- 11891526 TI - Evaluation of the malignant potential of aberrant crypt foci by immunohistochemical staining for beta-catenin in inflammation-induced rat colon carcinogenesis. AB - We previously showed that colitis enhanced the development of cancer and aberrant crypt foci (ACF) in 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in Fischer 344 rats. In this study, we examined the effect of two different anti inflammatory drugs [non-steroidal anti-inflammatory drugs (NSAIDs: Fenbufen) and a platelet activating factor-receptor antagonist (PAF-RA)] on the inflammation induced rat colon carcinogenesis. Furthermore, we examined the expression and the localization of beta-catenin protein, and the proliferating cell nuclear antigen (PCNA)-labeling index (LI) in ACF and cancer. PAF-RA significantly decreased the incidence of ACF in the rats (p<0.05), but Fenbufen did not affect the incidence of ACF and cancer. In most of the ACF (91%), beta-catenin was localized at the cell membrane like in normal colon epithelium. In about 9% of the ACF, beta catenin was overexpressed not only on the cell membrane but also in the cytoplasm. In all of the cancer cells, beta-catenin was overexpressed in the nucleus. When we compared the PCNA-LI in the ACF showing normal beta-catenin expression pattern with that in the ACF showing abnormal beta-catenin expression pattern (overexpression in cytoplasm), there was no significant difference of the PCNA-LI in these two different types of ACF. These findings suggest that immunohistochemical staining of ACF for beta-catenin can evaluate the malignant potential of ACF, and that PAF-RA can be used for preventing the development of ACF in inflammation-induced carcinogenesis. PMID- 11891528 TI - Expression of human SOX7 in normal tissues and tumors. AB - SOX transcription factors with high-mobility-group DNA-binding domain (HMG box) play key roles in embryogenesis. Some members of the SOX family are negative regulators of the WNT-beta-catenin-TCF signaling pathway. We have previously cloned and characterized human SOX17, constituting a subfamily with SOX7 and SOX18. Another group mapped SOX7 gene to human chromosome 8p22, and reported almost ubiquitous expression of 5.0-kb SOX7 mRNA in human normal tissues. Here, expression of SOX7 mRNA was investigated by using SOX7 specific probe, which hybridized to 3.8-kb human SOX7 mRNA, but not to 5.0-kb mRNA. SOX7 mRNA was relatively highly expressed in adult lung, trachea, lymph node, placenta, fetal lung, and heart. In adult heart, SOX7 mRNA was more highly expressed in ventricules, inter-ventricular septum and apex than in atriums. SOX7 mRNA was significantly up-regulated in pancreatic cancer cell lines BxPC-3, PSN-1, Hs766T, and in 4 cases out of 8 cases of primary gastric cancer. SOX7 mRNA was relatively highly expressed in a gastric cancer cell line MKN45, esophageal cancer cell lines TE2, TE3, TE4, TE5, TE7, TE8, TE11, TE12, and TE13. On the other hand, SOX7 mRNA was significantly down-regulated in 7 out of 18 cases of primary colorectal tumors, in 4 out of 9 cases of primary breast cancer, in 4 out of 14 cases of primary kidney tumors, and also in some cases of primary lung and prostate cancer. SOX7 gene might be one of cancer-associated genes on human chromosome 8p22. PMID- 11891529 TI - NSAID zaltoprofen improves the decrease in body weight in rodent sickness behavior models: proposed new applications of NSAIDs (Review). AB - In infectious diseases and during inflammation, anorexia, loss of body weight, malaise, fatigue and depression are induced. These symptoms are correctively called 'sickness behaviors', and the central actions of cytokines play a role in their induction. The loss of body weight in cancer cachexia is also a result of development of sickness behaviors. It has been reported that the administration of NSAID ibuprofen to patients with cancer cachexia improves the loss in body weight. We studied the effect of NSAID on the loss of body weight by using rodent sickness behavior models. We have reported that sickness behaviors such as anorexia, decrease in body weight, and loss of locomotor activity are induced in concanavalin A (Con A)-induced mouse hepatitis and carbon tetrachloride-induced rat hepatitis. Zaltoprofen is a non-steroidal anti-inflammatory drug (NSAID) causes potent inhibition of cyclooxygenase-2 with fewer side effects on the gastrointestinal tract. Zaltoprofen improves the loss in body weight in both Con A-treated mice and carbon tetrachloride-treated rats. These results suggest the possible application of zaltoprofen for the treatment of sickness behaviors including loss of body weight occurring in cancer cachexia. PMID- 11891530 TI - Expression of p57/Kip2 protein in normal and neoplastic thyroid tissues. AB - p57 (Kip2) belongs to the Cip/Kip family and is one of the universal negative regulators of the cell cycle. In this study, we investigated the p57 expression of various types of thyroid neoplasm. p57 overexpression was observed in only 4.2% of normal thyroid tissues. In follicular adenoma and minimally invasive follicular carcinoma, p57 was overexpressed in 100% and 91.7% of the cases, respectively. However, its incidence was significantly lower (p<0.0001) in widely invasive follicular carcinoma, of which only 36.4% overexpressed p57. This phenomenon was seen in 63.1% of papillary carcinoma and 13.3% of anaplastic (undifferentiated) carcinoma. Furthermore, poorly differentiated and undifferentiated carcinoma more frequently lacked p57 expression (p<0.0001). These results suggest that the down-regulation of p57 may play a role in the dedifferentiation of thyroid carcinoma and in follicular carcinoma mutating to be more invasive. PMID- 11891531 TI - Axin, the main component of the Wnt signaling pathway, is not mutated in kidney tumors in children. AB - The Wnt signaling pathway is essential for embryonic development and can be involved in tumorigenesis when aberrantly activated. In a subset of Wilms' tumors, beta-catenin mutations have been identified, and this suggested that abnormally activated Wnt signaling may contribute to tumorigenesis of this tumor. Because Axin has been recognized as a main component of Wnt signaling, and its mutations were reported in several types of malignancies, we analyzed Axin gene mutations in 22 pediatric renal tumors. Twenty-four sets of the primers, which cover the whole coding region of the Axin gene, were used for PCR-SSCP analyses. Samples revealing aberrant band patterns were further analyzed for sequencing. We have only identified 4 silent mutations in the coding region and 3 intronic polymorphisms in Axin gene, accordingly no pathogenetic gene mutations were detected. Our results indicated that mutations of Axin gene as a mechanism of tumorigenesis are not associated with pediatric renal tumors including Wilms' tumors. PMID- 11891532 TI - Metabolism of D-[3-3H]glucose, D-[5-3H]glucose, D-[U-14C]glucose, D-[1 14C]glucose and D-[6-14C]glucose in pancreatic islets in an animal model of type 2 diabetes. AB - This study aims at exploring specific aspects of D-glucose metabolism, so far not yet investigated, in pancreatic islets from adult control rats and animals (STZ rats) injected with streptozotocin during the neonatal period. The latter animals, which represent a current model of type-2 diabetes, displayed a lower body weight, higher plasma D-glucose concentration and lower insulinogenic index than control rats. The protein, DNA and insulin content were all also lower in islets prepared from STZ, rather than control rats. In the presence of 10.0 mM D glucose, the paired ratio between D-[U-14C]glucose oxidation and D-[5-3H]glucose utilization was also decreased in the islets from STZ rats. No significant difference between control and STZ rats was observed, however, in terms of the ratios between D-[3-3H]glucose and D-[5-3H]glucose utilization, between the generation of radioactive lactate from 14C-labelled D-glucose and tritiated D glucose utilization and between D-[1-14C]glucose and D-[6-14C]glucose oxidation. These findings reinforce the view that the previously documented preferential impairment of the oxidative modality of glycolysis in islets from STZ rats contrasts with the absence of any major anomaly in other variables of D-glucose catabolism. PMID- 11891533 TI - Mitochondria, calcium and nitric oxide in the apoptotic pathway of esophageal carcinoma cells induced by As2O3. AB - In order to explore the early apoptotic signal messengers and to search the apoptotic pathway, the morphological and functional changes of mitochondria were examined, and nitric oxide (NO) and calcium ions (Ca2+) were measured in the course of apoptosis in esophageal carcinoma cells induced by As2O3. The esophageal carcinoma cell line SHEEC1, established by HPV in synergy with TPA in our laboratory, were cultured with serum-free medium in a culture flask, 24-well plate and small culture chambers, and added with As2O3 at 1, 3, 5 micromol/l. After 0, 2, 4, 8, 12, 24 h of drug adding the NO were measured from extracellular cultured medium and the SHEEC1 cells were collected from flasks for electron microscopic examination. Fluorescent intensity (FI) of rhodamine 123 (Rho123) labeled cells was detected using laser confocal scanning microscope (LCSM) for evaluation of mitochondrial membrane potential. Intracellular Ca2+ of cells in small culture chambers labeled with Fluo-3 dye were measured using LCSM over time. After adding As2O3, SHEEC1 cells revealed characteristic morphological and functional changes of mitochondria such as hyperplasia, swelling and disruption, accompanying decrease of transmembrane potential (FI of Rho123 decreased). The Ca2+ level increased at once after adding As2O3 and the NO concentration increased step by step till 24 h, then apoptotic morphology of cells occurred. The results suggest that by inducement of As2O3 increasing Ca2+ and NO, the apoptotic signal messengers, will initiate the mitochondria-dependent apoptotic pathway. PMID- 11891534 TI - Suppressive effects of a new anti-inflammatory agent, IS-741, on dextran sulfate sodium-induced experimental colitis in rats. AB - A novel anti-inflammatory drug, IS-741, blocked the adhesion of inflammatory cells to microvascular endothelial cells in vivo and in vitro. We examined the efficacy of IS-741 in a dextran sulfate sodium (DSS)-induced colitis model. DSS colitis was induced by the oral administration of 3% DSS for 10 days in rats. The rats were then divided in two groups: a 1% DSS plus IS-741 group and a 1% DSS plus water group. IS-741 was dissolved in water and administered orally (10 mg/kg) once per day for 14 days. The rats treated with DSS plus IS-741 remained healthy, and their body weight increased. The wet weight of the colon was significantly lower and the total colon length was significantly longer in the IS 741-treated group. Histological examinations revealed a marked infiltration of inflammatory cells into both the mucosa and submucosa in the DSS plus water group, but these changes were attenuated in the IS-741-treated group. The mucosal damage score was significantly reduced by treatment with IS-741. IS-741 also significantly reduced the mucosal myeloperoxidase activity. FACS analysis revealed that IS-741 significantly reduced Mac-1 expression on blood neutrophils. In conclusion, IS-741 suppressed DSS-induced experimental colitis in rats. Some of the action of IS-741 may be associated with its inhibitory effects on the Mac 1 expression of neutrophils in association with the blockade of their adhesion to endothelial cells. The findings in this study suggest that IS-741 may be a useful new therapeutic agent for inflammatory bowel disease. PMID- 11891535 TI - Transactivation of human alpha-fetoprotein gene by X-gene product of hepatitis B virus in human hepatoma cells. AB - The X-gene product of hepatitis B virus (HBX) modulates a variety of viral and cellular genes relevant to hepatocarcinogenesis, where alpha-fetoprotein (AFP) is produced by hepatoma cells. In the present study, the possible mechanism by which HBX regulates AFP expression was investigated using three human hepatoma cells, HepG2, HuH-7 and Hep3B, which are known to contain the wild-type, the mutant-type and the deletion of p53, respectively. Transfection with the HBX expression vector stimulated the co-transfected AFP reporter gene expression in HepG2 cells and HuH-7 cells, but not in Hep3B cells. Transfection with the p53 expression vector repressed the AFP reporter gene expression in all three hepatoma cells, while overexpression of HBX counteracted the p53-induced repression. In addition, a G-->A substitution at nucleotide -119 in the AFP promoter sequence abrogated the stimulatory effect of HBX on the AFP promoter in HepG2 cells. These results suggest that HBX interacts with p53 to up-regulate AFP gene transcription probably by restoration of the p53-mediated repression of the AFP promotor activity. PMID- 11891536 TI - Adoptive immunotherapy of mucin1 expressing adenocarcinomas with mucin1 stimulated human peripheral blood mononuclear cells. AB - Mucin1 stimulated hematopoietic mononuclear cells (M1SHMC) from patients with breast cancer, adoptively transferred to non-obese diabetic, severe combined immunodeficient (NOD SCID) mice, extended survival in a therapy model of gross adenocarcinoma and prevented tumor growth in a model of minimal disease. M1SHMC exhibited specific lysis of a human breast adenocarcinoma cell line expressing mucin1, MCF-7 and produced interferon gamma. M1SHMC were injected intraperitoneally (IP) in NOD SCID mice after gross, palpable tumors appeared after MCF-7 were injected subcutaneously (SC). Survival was increased as compared to no M1SHMC controls. However tumors eventually regrew in all mice. To determine whether minimal disease (MD) could be controlled, NOD SCID were injected with MCF 7 cells, and on the same day, injected IP with M1SHMC. The M1SHMC injected mice were protected from tumor growth. These results imply that M1SHMC can prolong survival, but not cure NOD SCID mice bearing gross palpable adenocarcinomas. However in a minimal disease model tumor growth was prevented. PMID- 11891537 TI - A region close to Tp53 shows LOH in familial breast cancer. AB - The proportions of mutation of BRCA1 and BRCA2 detected in familial breast cancer vary in different regions. Most breast cancer families in Sweden cannot be explained by mutations in the known major susceptibility genes. Our previous studies have found a high frequency of LOH in the Tp53 region in familial breast cancer suggesting a putative tumor suppressor gene in this region, and the Tp53 gene was excluded as predisposing gene in these families by mutation screening. In order to identify other candidate tumor suppressor genes responsible for familial breast cancer, we performed LOH analysis in 98 paired tumor and blood samples from 91 breast cancer families using 11 microsatellite markers on chromosome 17p. Two loci with high frequency of LOH were found. One spanned the Tp53 gene, the other was distal to Tp53. Linkage studies were performed on 17p with 11 microsatellite markers in 102 breast cancer families with no detectable mutations in the BRCA1 and BRCA2 gene. The linkage analysis did not further support any of the regions suggested by the LOH study. However, since the Tp53 gene is already known to predispose to breast cancer as well as being involved in tumor progression, it is possible that also this region, close but distal to Tp53 contains a gene involved in familial and/or sporadic breast cancer development similar to Tp53. PMID- 11891539 TI - Molecular characterization of head and neck tumors by analysis of telomerase activity and a panel of microsatellite markers. AB - Head and neck cancer is a frequent malignancy with a complex, and up to now not clear etiology. The reactivation of telomerase activity and losses or gains of specific chromosomal regions, which point to deletions of tumor suppressor genes or amplification of oncogenes are supposed to be the molecular processes during the development and progression of head and neck cancer. Therefore, we analyzed telomerase activity and microsatellite markers using a genome wide panel of 28 microsatellite markers in 38 head and neck squamous-cell carcinomas (HNSCC). Our microsatellite marker set included distinct chromosomal areas that all likely harbor genes contributing to the carcinogenesis of HNSCC. DNA or protein lysates were obtained from primary tumors and compared to peripheral lymphocytes or corresponding normal tissue. At least one genomic alteration [loss of heterozygosity (LOH), or microsatellite instability (MSI)] was found in 31 of the 38 cases (82%). Most frequently we detected an LOH in the chromosomal region 9p12 21 where at least the tumor suppressor genes (TSG) p16INK4A, p14ARF and p15INKB are localized. The comparison between grade two and grade three tumors showed a highly changed frequency of LOH in the chromosomal region 7q31, where a putative TSG is predicted. Telomerase activity was present in 31/37 (83.8%) tumor samples independent of the histopathological staging and grading of the tumors. These molecular characterizations of HNSCC may be a further hint for the involvement of additional, so far unknown, TSGs in the tumor progression and will elucidate the regulation of telomerase. PMID- 11891538 TI - The environmental toxin 2,3,7,8-tetrachlorodibenzo-p-dioxin induces cytochrome P450 activity in high passage PC 3 and DU 145 human prostate cancer cell lines. AB - The study was conducted to investigate whether 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) induces cytochrome P450 (CYP) 1A1 and CYP1B1 via the aryl hydrocarbon receptor (AhR) in the hormone-independent human prostate cancer cell lines PC 3 and DU 145. No quantitative differences in the expression of AhR and its partner transcription factor ARNT were seen in low and high passage number PC 3 and DU 145 cells in the absence and presence of TCDD as assessed by RT-PCR and Western blotting. However, CYP1A1/1B1 activity, measured by the 7-ethoxyresorufin O-deethylase (EROD) assay, was induced by 10 and 100 nM TCDD only in high passage number PC 3 and DU 145 cells (PC 3, 7.7- and 2-fold stimulation; DU 145, 8.5- and 19.7-fold stimulation). Besides stimulation of EROD activity, induction of the expression of CYP1A1 and, to a lesser extend, of CYP1B1 by TCDD was also demonstrated by RT-PCR and Western blotting. However, 1-100 nM TCDD did not significantly alter cell cycle distribution and cell growth for up to five days. The induction of CYP1A1 and CYP1B1 by TCCD in the hormone-independent prostate cancer cell lines suggests that CYP induction should be considered in patients with advanced prostate cancer. This could result in higher elimination rates of concomitant drugs metabolized by these particular CYP isoenzymes. PMID- 11891540 TI - Response of the immune system of mammary tumor-bearing rats to cyclophosphamide and soluble low-molecular-mass tumor-associated antigens: rate of lymphoid infiltration and distribution of T lymphocytes in tumors. AB - We have shown previously that soluble low-molecular-mass tumor-associated antigens (sTAA) promote the anti-tumor effect of the anticancer drug cyclophosphamide (CPA) on rat mammary carcinogenesis. In this study, we analyzed the possible mechanism underlying this phenomenon. Studies were performed on tumors obtained from the following groups of mammary tumor-bearing rats: i) control rats, ii) rats treated with sTAA, iii) rats treated with CPA, iv) rats treated with CPA and sTAA. All analyzed tumors represented different types of invasive duct carcinomas. The rate of lymphoid infiltration and T cell content (CD4+ and CD8+ cells) of tumors were analyzed immunohistochemically. In parallel, mitotic index was evaluated in tumor cells. In tumor-bearing rats, high lymphoid proliferation was found at the periphery of tumors, and to a lesser extent deep inside the tumors. In control tumors, CD4+ T cell content was very low whereas CD8+ cells were highly abundant, especially at the tumor periphery. Treatment with sTAA significantly increased the total number of lymph cells and the number of CD8+ lymphocytes inside the tumors. Cytoplasmic vacuolization, decreased mitotic index and various degrees of fibrosis were the most distinct changes in tumors treated with CPA alone. CPA also sharply decreased the activity of all lymph cells studied, especially of CD4+ lymphocytes which could no longer be observed following this treatment. The combined treatment of CPA and sTAA increased the number of lymph cells, although they did not reach control levels. Inhibition of mainly CD4+ lymphocyte synthesis of CPA was confirmed by the low CD4/CD8 ratio, which increased slightly after the combined treatment with CPA and sTAA. Findings in the present study demonstrate that vaccination with sTAA actively promotes the generation of the host's antitumor immune response. PMID- 11891542 TI - Hans Ussing memorial issue: epithelial membrane transport. PMID- 11891543 TI - Hans Ussing, experiments and models. AB - The article describes work and impact of Hans H. Ussing, a founder of epithelial physiology. Emphasis is on Ussing's model of epithelial transport, which showed early how a complex function can arise from a few basic principles. The KJU-model was developed 1958 for the amphibian epidermis and later applied and adapted to many epithelia, but especially to those that express amiloride-sensitive sodium channels in their apical membrane. Some of the subsequent research dealing with such channels and their cellular environment is briefly reviewed. The ideas of Hans Ussing were and are an inspiration to many of us, who continue to work in the way Ussing has taught us. PMID- 11891544 TI - Ussing's two-membrane hypothesis: the model and half a century of progress. PMID- 11891545 TI - The influence of Ussing's pioneering work on a Dutch physiologist or how I stopped worrying. PMID- 11891546 TI - Remembrances of renal potassium transport. PMID- 11891547 TI - Intestinal absorption of hexoses and amino acids: from apical cytosol to villus capillaries. PMID- 11891548 TI - The paracellular shunt of proximal tubule. PMID- 11891549 TI - Role of mitochondria-rich cells for passive chloride transport, with a discussion of Ussing's contribution to our understanding of shunt pathways in epithelia. AB - A non-invasive method is applied for studying ion transport by single isolated epidermal mitochondria-rich (MR) cells. MR cells of toad skin (Bufo bufo) were prepared by trypsin (or pronase) treatment of the isolated epithelium bathed in Ca2+-free Ringer. Glass pipettes were pulled and heat- polished to obtain a tip of 2-4 mm with parallel walls and low tip resistances. The neck of an MR-cell was sucked into the tip of the pipette for being 'clamped mechanically' by the heat polished glass wall. In this configuration the apical cell membrane faces the pipette solution while the major neck region and the cell body are in the electrically grounded bath. With Ringer in bath and pipette, transcellular voltage clamp currents were composed of an ohmic (I(leak)) and a dynamic (I(dynamic)) component. The dynamic component was studied by stepping the transcellular potential (Vp) from a holding value of +50 mV to the hyperpolarizing region (50 > Vp > or = -100 mV). The steady state I(dynamic)-Vp relationship was strongly outward rectified with I(dynamic) being practically zero for Vp > 0 mV. At Vp = -100 mV, MR cells isolated by trypsin or pronase generated a steady-state I(dynamic) of,-2.72 +/- 0.40 nA/cell (N = 21 MR cells). Continuous superfusion of the MR cell during recording increased the current to 7.99 +/- 1.48 nA/cell (N = 10 MR cells). The time course of the reversible activation of G(dynamic) varied among cells, but was usually sigmoid with T1/2 decreasing with Vp (-25 > or = Vp > or = -100 mV). T1/2 was in the order of 10 sec at Vp = -100 mV. The single-MR-cell currents recorded in this study are fully compatible with Cl- currents estimated by relating density of MR cells to transepithelial ICl or by measurements with the self-referencing ('vibrating') probe technique. In the discussion, Ussing's work on epithelial shunt pathways is considered. His thinking and experiments leading to his theory of isotonic transport in leaky epithelia is emphasized. It is our thesis that the understanding of the physiology of epithelia owes as much to Ussing's studies of shunt pathways as to his studies of the active sodium pathway. PMID- 11891550 TI - Sodium absorption, volume control and potassium channels: in tribute to a great biologist. AB - It is well established, for all Na-absorbing epithelia, that an increase in the rate of transcellular Na+ absorption is accompanied by an increase in the conductance of the basolateral membrane to K+. For the case of small intestinal epithelial cells from the salamander Necturus maculosus, where the rate of transcellular Na+ absorption can be increased manyfold by the addition of sugars or amino acids to the luminal bathing solution, it appears that this parallelism between Na-K pump rate and basolateral membrane K+ conductance is closely related to volume regulation by the enterocyte. Recent studies have disclosed the presence of stretch-activated K+ channels, in a highly enriched basolateral membrane fraction isolated from these epithelial cells, whose activity is increased by an increase in vesicle volume and inhibited by a decrease in vesicle volume or ATP. The activity of this channel also appears to be regulated by the degree of organization of the cortical actin cytoskeleton; activity is increased by depolymerization of the actin cytoskeleton and decreased by repolymerization of that structure. We postulate that the inhibitory effect of ATP is related to its role in promoting the polymerization of G-actin to form F-actin. We propose that enterocyte swelling that results from the intracellular accumulation of sugars or amino acids in osmotically active forms brings about disorganization of the cortical actin cytoskeleton and activates these channels and is, at least in part, responsible for the "pump-leak parallelism" in this amphibian. PMID- 11891551 TI - The modulation of ionic currents in excitable tissues by n-3 polyunsaturated fatty acids. PMID- 11891552 TI - Specific lectin binding to beta1 integrin and fibronectin on the apical membrane of madin-darby canine kidney cells. AB - Although lectins have previously been used to identify specific cell types in the kidney and various other tissues, the proteins labeled were not identified. We hypothesized that fluorescently labeled lectins could provide a useful tool for direct labeling of membrane-associated glycoproteins. Protein fractions from Madin-Darby canine kidney (MDCK) cells were exposed to a panel of 16 fluorescently labeled lectins to identify suitable lectin-protein pairs. Peanut agglutinin (PNA) selectively bound a 220-240 kDa O-linked glycoprotein with a slightly acidic isoelectric point, while Sambucus nigra agglutinin (SNA) labeled a 130 kDa glycoprotein with a highly acidic isoelectric point. Both proteins were readily labeled by lectins applied to the apical surface of living confluent cells. The proteins were isolated by lectin affinity columns and identified by mass spectrometry. Peptides from the PNA-binding protein shared molecular weight and amino acid composition with fibronectin. Fragments of the SNA-binding protein showed amino-acid identity with peptides from beta1 integrin. The identities of these proteins were validated by Western blotting. Binding of PNA to a 220 kDa protein was inhibited by an anti-fibronectin antibody, and binding of a 130 kDa protein by SNA was diminished by an anti-beta1 integrin antibody. We conclude that PNA and SNA can be used as specific markers for fibronectin and beta1 integrin, respectively, in MDCK cells. PMID- 11891553 TI - Fluid secretion in Rhodnius upper malpighian tubules (UMT): water osmotic permeabilities and morphometric studies. AB - We have measured the osmotic permeability of the basolateral cell membrane (Poscb) and compared it with the transepithelial permeability (Poste) to calculate the paracellular (Posp) permeability of the upper malpighian tubules (UMT) of the 5th instar of Rhodnius prolixus under several experimental conditions, namely, at rest and after stimulation to secrete with 5-HT, each under control conditions (no treatment), after treatment with pCMBS, and after addition of pCMBS and DTT. Secretion rate is negligible at rest. During stimulation mean secretion rate is 43.5 nl/cm2 sec. Secretion is severely curtailed by pCMBS and fully restored by DTT. Poscb = 9.4 (resting, control); 5.8 (control + pCMBS); 10.7 (control + pCMBS + DTT); 20.6 (stimulated, control); 14.7 (stimulated + pCMBS); 49.1 (stimulated + pCMBS + DTT) (x10?4 cm3/cm2 sec Osm). Calculated Posp are higher than the transcellular permeability, Posc, at rest and after stimulation. Electron micrograph morphometry of UMT sections show that cells significantly decrease their volume after stimulation. Lateral intercellular space (LIS) and basolateral extracellular labyrinth (BEL) are barely discernible at rest. LIS and BEL are widely dilated in stimulated UMT. Thus, ions have restricted access to the deep and narrow basolateral cell membrane indentations at rest, but they have ready access to cell membrane indentations after stimulation, because of the opening of LIS and BEL. These findings are discussed in relation to isosmotic secretion. The rate-limiting step for paracellular movement is located at the smooth septate junctions. PMID- 11891554 TI - Effect of the putative Ca2+-receptor agonist Gd3+ on the active transepithelial Na+ transport in frog skin. AB - In this communication we show that Gd3+ acts as an activator of the apical sodium channel (ENaC) in frog skin epithelia. Application of Gd3+ to the apical solution of frog skin epithelia increased the Na+ absorption measured as the amiloride inhibitable short-circuit current (Isc). The stimulation was dose dependent with a concentration for half-maximal stimulation (EC50) of 0.023 mM. The change in Isc was found to correlate with the net Na+ flux, confirming that Gd3+ enhances Na+ absorption. By monitoring the cellular potential (Vsc) with microelectrodes during addition of Gd3+, it was found that Vsc depolarized as Isc rose, indicating that Gd3+ affects apical Na+ permeability (PNa). This was confirmed by measuring the I/V relations of the apical membrane. In the presence of benzimidazolylguanidin (BIG), a drug known to abolish the Na+ self-inhibition, Gd3+ had no effect on Isc. The Na+ self-inhibition was investigated using fast changes of the apical Na+ concentration on K+-depolarized epithelia. BIG was found to abolish the Na+ self-inhibition and to activate the basal Na+ transport, whereas Gd3+ only activated the basal Na+ transport but had no effect on the self inhibition. These results indicate the existence of an alternative nonhormonal mechanism to Na+ self-inhibition, via which both Gd3+ and BIG act, possibly components of the Na+ feedback inhibition system. PMID- 11891555 TI - The polarized distribution of Na+, K+-ATPase and active transport across epithelia. PMID- 11891556 TI - Intracellular pH as a regulator of Na + transport. AB - Na reabsorption by tight epithelia, such as frog skin and toad urinary bladder, is highly sensitive to the acid-base status of the cytoplasm. This can be observed in intact epithelia by acidifying the intracellular compartment with acute hypercapnia. Both apical membrane Na channels, which are responsible for the uptake of Na into the cell, and basolateral membrane K channels, which are required for there cycling of K that is actively transported into the cell through the Na/K pump, are shut down by low intracellular pH. This suggests the possibility that cell pH may serve as an important regulator of transport. One possible role is as a second messenger for rapid effects of the adrenal mineralocorticoid aldosterone. PMID- 11891557 TI - Mechanisms of aldosterone's action on epithelial Na + transport. AB - Aldosterone maintains total organism sodium balance in all higher vertebrates. The level of sodium reabsorption is primarily determined by the action of aldosterone on epithelial sodium channels (ENaC) in the distal nephron. Recent work shows that, in an aldosterone-sensitive renal cell line (A6), aldosterone regulates sodium reabsorption by short- and long-term processes. In the short term, aldosterone regulates sodium transport by inducing expression of the small G-protein, K-Ras2A, by stimulating the activity of methyl transferase and S adenosyl-homocysteine hydrolase to activate Ras by methylation, and, possibly, by subsequent activation by K-Ras2A of phosphatidylinositol phosphate-5-kinase (PIP 5-K) and phosphatidylinositol-3-kinase (PI-3-K), which ultimately activates ENaC. In the long term, aldosterone regulates sodium transport by altering trafficking, assembly, and degradation of ENaC. PMID- 11891558 TI - The pump and leak steady-state concept with a variety of regulated leak pathways. AB - The paper will reflect on how Ussing has affected my own scientific work and how he created much of the framework within which I have been working. I have used five examples: (i) The first description of a 1:1 exchange diffusion was introduced by Ussing in 1947 and has been found to be of great physiological significance in most cells. We found that Cl- transport in Ehrlich ascites tumor cells (EATC) was completely dominated by an exchange diffusion process, as defined by Ussing, and, thus, the Cl- conductance was much lower than previously estimated from measurements of unidirectional tracer fluxes. This had a major influence on my later description of a swelling-activated Cl- conductance. (ii) The pump-leak steady-state concept for cell volume control was introduced by Krogh in 1946, but it was developed in detail by Leaf and Ussing in 1959. This concept was the basis for me and others, when we later found that the passive ion leaks play an active role in cell volume control. (iii) The use of isotopes and Ussing's famous flux ratio equation provided an ingenious instrument for distinguishing the various transport routes. We used this to identify the Na,K,2Cl cotransport system as accounting for maintaining a [Cl-]i in the EATC far above thermodynamic equilibrium, as well as accounting for the ion uptake during a regulatory volume increase (RVI) in EATC, similar to what Ussing had found in frog skin. (iv) Short-circuit current setup in the Ussing chamber is still used in laboratories around the world to study ion transport across epithelia. A few results on Cl- transport across the operculum epithelium of the small eurohaline fish Fundulus heteroclitus mounted in small Ussing chambers are presented. (v) Shrinkage-activated Na+ conductance and its possible role in isotonic secretion in frog skin glands is finally discussed. PMID- 11891559 TI - Modulation of intracellular calcium-release channels by calmodulin. PMID- 11891560 TI - Functional characterization of CitM, the Mg2+-citrate transporter. AB - The CitM transporter from Bacillus subtilis transports citrate as a complex with Mg2+. In this study, CitM was functionally expressed and characterized in E. coli DH5a cells. In the presence of saturating Mg2+ concentrations, the Km for citrate in CitM was 274 mM, similar to previous studies using whole cells of B. subtilis. CitM has a high substrate specificity for citrate. Other di- and tricarboxylic acids including succinate, isocitrate, cis-aconitate and tricarballylic acid did not significantly inhibit the uptake of citrate in the presence of Mg2+. However, CitM accepts complexes of citrate with metal ions other than Mg2+. The highest rate of citrate transport was seen in the presence of Mg2+, followed in order of preference by Mn2+, Ba2+, Ni2+, Co2+ and Ca2+. Citrate transport by CitM appears to be proton coupled. The transport was inhibited in transport buffers more alkaline than pH 7.5 and not affected by pH at acidic values. Transport was also inhibited by ionophores that affect the transmembrane proton gradient, including FCCP, TCC and nigericin. Valinomycin did not affect the uptake by CitM, suggesting that transport is electroneutral. In conclusion, the cloned CitM transporter from B. subtilis expressed in E. coli has properties similar to the transporter in intact B. subtilis cells. The results support a transport model with a coupling stoichiometry of one proton coupled to the uptake of one complex of (Mg2+-citrate)1-. PMID- 11891561 TI - Expression and cellular distribution pattern of plasma membrane calcium pump isoforms in rat pancreatic islets. AB - This work is aimed at identifying the presence and cellular distribution pattern of plasma membrane calcium pump (PMCA) isoforms in normal rat pancreatic islet. Microsomal fractions of isolated islets and exocrine tissue were analyzed to detect different PMCA isoforms. The cellular distribution pattern of these PMCAs in the islets was also studied in fixed pancreas sections incubated with antibodies against PMCAs and insulin. Antibody 5F10, which reacts with all PMCA variants, showed multiple bands in the blots in the 127-134 kDa region, indicating the presence of several isoforms. Microsomes also reacted positively with specific antibodies for individual PMCA isoforms, generating a band of the expected size. Antibody 5F10 immunocytochemically labeled the plasma cell membrane of both b- and non-b-cells, but predominantly the former. All islet cells were also labeled with antibodies against isoforms 1 and 4, while the antibody reacting with isoform 3 labeled exclusively b-cells. A few b- and non-b cells were positively labeled with the antibody reacting with PMCA b variant. Negative results were obtained with the antibody against isoform 2. Further studies, together with previous reports on the modulatory effect of insulin secretagogues and blockers upon PMCA activity, may provide evidence of the importance of this particular PMCA expression for islet function under normal and pathological conditions. PMID- 11891562 TI - A chi(2) test for model determination and sublevel detection in ion channel analysis. AB - A chi(2) test is proposed that provides a means of discriminating between different Markov models used for the description of a measured (patch clamp) time series. It is based on a test statistic constructed from the measured and the predicted number of transitions between the current levels. With a certain probability, this test statistic is below a threshold if the model with a reduced number of degrees of freedom is compatible with the data. A second criterion is provided by the dependence of the test statistic on the number of data points. For data generated by the alternative model it increases linearly. The applicability of this test for verifying and rejecting models is illustrated by means of time series generated by two distinct channels with different conductances and by time series generated by one channel with two conductance levels. For noisy data, a noise correction is proposed, which eliminates noise induced false jumps that would interfere with the test. It is shown that the test can also be extended to aggregated Markov models. PMID- 11891563 TI - Membrane dynamics in the malpighian tubules of the house cricket, acheta domesticus. AB - In Acheta domesticus, the Malpighian tubules (Mt) are composed of three morphologically distinct regions (proximal, mid and distal), each consisting of a single cell type. The bulk of the Mt is composed of the midtubule, which shows the greatest response to corticotropin releasing factor-related diuretic peptides (CRF-DP). We know from previous laboratory studies that the second messenger cAMP and its analog dibutyryl cAMP (db-cAMP) cause an approximate doubling in the secretion rate and that this is accompanied by notable ultrastructural changes in the midtubule, especially membrane reorganization in the basal area and extensive vesiculation of the cytoplasm. In this study, we examined the morphological changes in membranes both at the cell surface and internally. By enzymatically removing the basal lamina, we examined the increase in spacing between infolded membranes initiated by db-cAMP stimulation. To examine the intracellular membranes, we used a technique developed for use in invertebrate tissues. This allowed the removal of the cytoplasm for high resolution scanning electron microscopy (HR-SEM) while maintaining the integrity of the lipid constituents of the cell. By using HR-SEM and confocal laser scanning microscopy (CLSM), we gained a unique three-dimensional perspective of the complexity of the internal membrane system of the A. domesticus Mt in both the unstimulated and db-cAMP stimulated states. PMID- 11891564 TI - Comparison of the osmolyte transport properties induced by trAE1 versus IClswell in Xenopus oocytes. AB - During cell swelling, cells release organic osmolytes via a volume-activated channel as part of the regulatory volume decrease. The erythrocyte membrane protein AE1 (band 3), has been shown to be involved in regulatory volume responses of fish erythrocytes. Previous studies showed that the expression of trout AE1 in Xenopus laevis oocytes induces band 3 anion exchange activity and organic osmolyte channel activity. However, an endogenous swelling-activated anion channel, IClswell, is present in Xenopus oocyte membranes. Therefore, it is not yet known whether a new organic osmolyte channel is formed or whether the endogenous channel, IClswell, is activated when trout AE1 is expressed in the oocytes. The purpose of this study was to determine whether the expression of trout AE1 in Xenopus oocytes leads to the formation and membrane insertion of a new organic osmolyte channel or activates IClswell. To differentiate between the two possibilities, we compared the time courses, pH profiles and inhibitor sensitivities of both trout AE1 and IClswell. The results of taurine-uptake experiments show that the time courses and pH levels for optimum expression of trout AE1 and IClswell differ significantly. The inhibitor sensitivities of the organic osmolyte channel mediated by trout AE1 and IClswell are also significantly different, strongly suggesting that the expression of trout AE1 in Xenopus oocytes does not activate IClswell, but rather forms a new organic osmolyte channel. PMID- 11891565 TI - Functional characterization of intestinal L-carnitine transport. AB - The carnitine transporter OCTN2 is responsible for the renal reabsorption of filtered L-carnitine. However, there is controversy regarding the intestinal L carnitine transport mechanism(s). In this study, the characteristics of L carnitine transport in both, isolated chicken enterocytes and brush-border membrane vesicles (BBMV) were studied. In situ hybridization was also performed in chicken small intestine. Chicken enterocytes maintain a steady-state L carnitine gradient of 5 to 1 and 90% of the transported L-carnitine remains in a readily diffusive form. After 5 min, L-Carnitine uptake into BBMV overshot the equilibrium value by a factor of 2.5. Concentrative L-carnitine transport is Na+ , membrane voltage-and pH-dependent, has a high affinity for L-carnitine (Km 26 - 31 microM ) and a 1:1 Na+: L-carnitine stoichiometry. L-Carnitine uptake into either enterocytes or BBMV was inhibited by excess amount of cold L-carnitine > D carnitine = acetyl-L-carnitine = gamma-butyrobetaine > palmitoyl-L-carnitine > betaine > TEA, whereas alanine, histidine, GABA or choline were without significant effect. In situ hybridization studies revealed that only the cells lining the intestinal villus expressed OCTN2 mRNA. This is the first demonstration of the operation of a Na+/L-carnitine cotransport system in the apical membrane of enterocytes. This transporter has properties similar to those of OCTN2. PMID- 11891566 TI - Chloride conductance determining membrane potential of rabbit articular chondrocytes. AB - Membrane conductance of cultured rabbit articular chondrocytes was characterized by means of the patch-clamp technique. The resting membrane potential of the articular chondrocytes was about -42 mV. The membrane potential shifted in accordance with the prediction by the Nernst equation for Cl- when intracellular and extracellular concentrations of Cl- were changed. On the other hand, change in extracellular concentration of K+ produced no shift in the membrane potential of chondrocytes. The Cl- channel blocker 4-acetamido-4'-isothiocyanatostilbene 2'2-disulfonic acid (SITS) depolarized the membrane potential. These findings suggest that the membrane potential of the chondrocytes is determined mainly by Cl- conductance. Using the cell-attached patch-clamp method, a large unitary conductance of 217 pS was observed in the articular chondrocytes. The unitary current was reversibly blocked by SITS. Therefore, the unitary current was carried by Cl-. The Cl- channel showed voltage-dependent activation and the channels exhibited long-lasting openings. Therefore, the membrane potential of rabbit cultured articular chondrocytes was mainly determined by the activities of the large-conductance and voltage-dependent Cl- channels. PMID- 11891567 TI - Electrophysiological behavior of the TolC channel-tunnel in planar lipid bilayers. AB - Escherichia coli TolC assembles into the unique channel-tunnel structure spanning the outer membrane and periplasmic space. The structure is constricted only at the periplasmic entrance of the tunnel and this must be opened to allow export of substrates bound by cognate inner membrane complexes. We have investigated the electrophysiological behavior of TolC reconstituted into planar lipid bilayers, in particular the influence of the membrane potential, the electrolyte concentration and pH. TolC inserted in one orientation into the membrane. The resultant pores were stable and showed no voltage-dependent opening or closing. Nevertheless, TolC could adopt up to three conductance substates. The pores were cation-selective with a permeability ratio of potassium to chloride ions of 16.5. The single-channel conductance was higher when the protein was inserted from the side with negative potential. It showed a nonlinear dependence on the concentration of the electrolyte in the bulk solution and decreased as the pH was lowered. The calculated pK of the apparent closing was 4.5. The electrophysiological characterization is discussed in relation to the TolC structure, in particular the periplasmic entrance. PMID- 11891568 TI - Inhibition of gap junction hemichannels by chloride channel blockers. AB - Electrophysiological methods were used to assess the effect of chloride-channel blockers on the macroscopic and microscopic currents of mouse connexin50 (Cx50) and rat connexin46 (Cx46) hemichannels expressed in Xenopus laevis oocytes. Oocytes were voltage-clamped at -50 mV and hemichannel currents (ICx50 or ICx46) were activated by lowering the extracellular Ca2+ concentration ([Ca2+]o) from 5 mM to 10 microM. Ion-replacement experiments suggested that ICx50 is carried primarily (>95%) by monovalent cations (PK : PNa : PCl = 1.0 : 0.74 : 0.05). ICx50 was inhibited by 18beta-glycyrrhetinic acid (apparent Ki, 2 microM), gadolinium (3 microM), flufenamic acid (3 microM), niflumic acid (11 microM), NPPB (15 microM), diphenyl-2-carboxylate (26 microM), and octanol (177 microM). With the exception of octanol, niflumic acid, and diphenyl-2-carboxylate, the above agents also inhibited ICx46. Anthracene-9-carboxylate, furosemide, DIDS, SITS, IAA-94, and tamoxifen had no inhibitory effect on either ICx50 or ICx46. The kinetics of ICx50 inhibition were not altered at widely different [Ca2+]o (10 500 microM), suggesting that drug-hemichannel interaction does not involve the Ca2+ binding site. In excised membrane patches, application of flufenamic acid or octanol to the extracellular surface of Cx50 hemichannels reduced single channel open probability without altering the single-channel conductance, but application to the cytoplasmic surface had no effect on the channels. We conclude that some chloride-channel blockers inhibit lens-connexin hemichannels by acting on a site accessible only from the extracellular space, and that drug-hemichannel interaction involves a high-affinity site other than the Ca2+ binding site. PMID- 11891569 TI - Immunolocalization of P2Y4 and P2Y2 purinergic receptors in strial marginal cells and vestibular dark cells. AB - K+ secretion by strial marginal cell and vestibular dark cell epithelia is regulated by UTP and ATP at both the apical and basolateral membranes, suggesting control by P2Y2 and/or P2Y4 purinergic receptors. Immunolocalization was used to determine the identity and distribution of these putative receptors. Membrane proteins from gerbil brain, gerbil vestibular labyrinth and gerbil stria vascularis were isolated and analyzed by Western blot. P2Y2 antibody stained one band at 42 kDa for each tissue, whereas P2Y4 antibody stained 3 bands on gerbil brain (75, 55 and 36 kDa), one band on gerbil stria vascularis (55 kDa) and two bands on vestibular labyrinth (42 and 56 kDa). All bands were absent when the antibodies were blocked with their respective antigenic peptide. P2Y4 was immunolocalized by fluorescence confocal microscopy to only the apical membrane of strial marginal cells and vestibular dark cells and was similar to apical immunostaining of KCNE1 in the same cells. By contrast, P2Y2 was observed on the basolateral but not the apical membrane of dark cells. Similarly, in the stria vascularis P2Y2 was observed in the basolateral region but not the apical membrane of marginal cells. Additional staining was observed in the spiral ligament underlying the stria vascularis. These findings identify the molecular bases of the regulation of K+ secretion by apical and basolateral UTP in the inner ear. PMID- 11891570 TI - Protein orientation affects the efficiency of functional protein transplantation into the xenopus oocyte membrane. AB - Xenopus oocytes incorporate into their plasma membrane nicotinic acetylcholine receptors (nAChRs) after intracellular injection of lipid vesicles bearing this protein. The advantage of this approach over the classical oocyte expression system lies in the transplantation of native, fully processed proteins, although the efficiency of functional incorporation of nAChRs is low. We have now studied the incorporation into the oocyte membrane of the Torpedo chloride channel (ClC 0), a minor contaminant protein in some nAChR preparations. nAChR-injected oocytes incorporated functional ClC-0: i) in a higher number than functional nAChRs; ii) retaining their original properties; and iii) with a right-side-out orientation in the oocyte membrane. In an attempt to elucidate the reasons for the low efficiency in the functional incorporation of nAChRs into the oocyte membrane, we combined electrophysiological and [125I]alpha-bungarotoxin-binding experiments. Up to 3% of injected nAChRs were present in the oocyte plasma membrane at a given time. Thus, fusion of lipoproteosome vesicles to the oocyte plasma membrane is not the limiting factor for an efficient functional transplantation of foreign proteins. Accounting for the low rate of functional transplantation of nAChRs is their backward orientation in the oocyte membrane, since about 80% of them adopted an out-side-in orientation. Other factors, including differences in the susceptibility of the transplanted proteins to intracellular damage should also be considered. PMID- 11891571 TI - Basis for intracellular retention of a human mutant of the retinal rod channel alpha subunit. AB - A mutant of the a subunit of the retinal rod cyclic GMP-gated channel, [Arg654(1 bp del)], corresponding to a truncated alphaR654Dstop subunit, was previously described in patients with retinitis pigmentosa: when expressed in HEK-293 cells, this mutated a subunit was retained inside the cell, but had normal channel activity in one case where it reached the plasma membrane, indicating that the mechanism of targeting is altered by the mutation, but not the function of the channel. The corresponding mutants of the bovine rod channel (alphaR656D stop), and of the closely related olfactory neuron channel (alphaR632Dstop) alpha subunits were expressed in Xenopus oocytes and their activity was analyzed by patch-clamp. Like their human homologue, these two channels have no activity, and we show that their GFP fusion proteins are accumulated into intracellular compartments. The truncation alone or the R/D mutation alone do not prevent or modify channel activity, indicating that neither the R656 residue nor the C terminal domain downstream of R656 is necessary for homomeric channel targeting and function. Several mutations of R656 and of the preceding residues in the R656Dstop mutant disclose that the motif responsible for the absence of channel activity is an endoplasmic reticulum retention signal (KXKXXstop) in which the nature of the residues in positions -1 and -4 is determinant. PMID- 11891573 TI - Mutation D384N alters recovery of the immobilized gating charge in rat brain IIA sodium channels. AB - Rat brain (rBIIA) sodium channel fast inactivation kinetics and the time course of recovery of the immobilized gating charge were compared for wild type (WT) and the pore mutant D384N heterologously expressed in Xenopus oocytes with or without the accessory beta1-subunit. In the absence of the beta1-subunit, WT and D384N showed characteristic bimodal inactivation kinetics, but with the fast gating mode significantly more pronounced in D384N. Both, for WT and D384N, coexpression of the beta1-subunit further shifted the time course of inactivation to the fast gating mode. However, the recovery of the immobilized gating charge (Qg) of D384N was clearly faster than in WT, irrespective of the presence of the beta1-subunit. This was also reflected by the kinetics of the slow Ig OFF tail. On the other hand, the voltage dependence of the Qg-recovery was not changed by the mutation. These data suggest a direct interaction between the selectivity filter and the immobilized voltage sensor S4D4 of rBIIA sodium channels. PMID- 11891572 TI - Cyclic AMP-dependent Cl secretion is regulated by multiple phosphodiesterase subtypes in human colonic epithelial cells. AB - The role of phosphodiesterase (PDE) isoforms in regulation of transepithelial Cl secretion was investigated using cultured monolayers of T84 cells grown on membrane filters. Identification of the major PDE isoforms present in these cells was determined using ion exchange chromatography in combination with biochemical assays for cGMP and cAMP hydrolysis. The most abundant PDE isoform in these cells was PDE4 accounting for 70-80% of the total cAMP hydrolysis within the cytosolic and membrane fractions from these cells. The PDE3 isoform was also identified in both cytosolic and membrane fractions accounting for 20% of the total cAMP hydrolysis in the cytosolic fraction and 15-30% of the total cAMP hydrolysis observed in the membrane fraction. A large portion of the total cGMP hydrolysis detected in cytosolic and membrane fractions of T84 cells was mediated by PDE5 (50-75%). Treatment of confluent monolayers of T84 cells with various PDE inhibitors produced significant increases in short-circuit current (Isc). The PDE3-selective inhibitors terqinsin, milrinone and cilostamide produced increases in Isc with EC50 values of 0.6 nM, 8.0 nM and 0.5 microM respectively. These values were in close agreement with the IC50 values for cAMP hydrolysis. The effects of the PDE1-(8-MM-IBMX) and PDE4-(RP-73401) selective inhibitors on Isc were significantly less potent than PDE3 inhibitors with EC50 values of >7 microM and >50 microM respectively. However, the effects of 8-MM-IBMX and terqinsin on Cl secretion were additive, suggesting that inhibition of PDE1 also increases Cl secretion. The effect of PDE inhibitors on Isc were significantly blocked by apical treatment with glibenclamide (an inhibitor of the CFTR Cl channel) and by basolateral bumetanide, an inhibitor of Na-K-2Cl cotransport activity. These results indicate that inhibition of PDE activity in T84 cells stimulates transepithelial Cl secretion and that PDE1 and PDE3 are involved in regulating the rate of secretion. PMID- 11891574 TI - Swelling-activated taurine and creatine effluxes from rat cortical astrocytes are pharmacologically distinct. AB - Primary cultures of rat cortical astrocytes undergo a swelling-activated loss of taurine and creatine. In this study, the pharmacological characteristics of the taurine and creatine efflux pathways were compared, and significant differences were shown to exist between the two. Both taurine and creatine effluxes were rapidly activated upon exposure of astrocytes to hypo-osmotic media, and rapidly inactivated upon their return to iso-osmotic media. The relative rates of taurine and creatine efflux depended upon the magnitude of the hypo-osmotic shock. Anion transport inhibitors strongly inhibited taurine efflux, with the order of potency being NPPB > DIDS > niflumic acid. DIDS and NPPB had less of an inhibitory effect on creatine efflux, whereas tamoxifen and niflumic acid actually stimulated creatine efflux. These data are consistent with separate pathways for taurine and creatine loss during astrocyte swelling. PMID- 11891575 TI - Distribution and regulation of ENaC subunit and CFTR mRNA expression in murine female reproductive tract. AB - The present study investigated the regional distribution and cyclic changes in the mRNA expression of epithelial Na+ channel (ENaC) subunit and cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated Cl- channel, in adult female mouse reproductive tract. In situ hybridization revealed that in contrast to the abundant expression of CFTR, ENaC (alpha, beta, gamma) mRNA signal was not detected throughout the estrus cycle in the ovary and oviduct. Messenger RNA for all ENaC subunits was abundantly detected in the cervical and vaginal epithelia throughout the estrus cycle but for CFTR, mRNA was found only at proestrus. In the uterine epithelium, alphaENaC mRNA was detected at diestrus but not found at any other stage, while CFTR mRNA was only detected at early estrus but not other stages. Semi-quantitative RT-PCR detected mRNA for all ENaC subunits in the uterus throughout the cycle with maximal expression at diestrus and CFTR mRNA was only found in the early stages of the cycle. The involvement of ENaC and CFTR in Na+ absorption and Cl- secretion was demonstrated in cultured endometrial epithelia using the short-circuit current technique and found to be influenced by ovarian hormones. Taken together, these data indicate a main secretory role of the ovary and oviduct and a predominantly absorptive role of the cervix and vagina. The present results also suggest an ability of the uterus to secrete and absorb at different stages of the estrus cycle. Variations in the fluid profiles may be dictated by the regional and cyclic variations in expression of ENaC and CFTR and are likely to contribute to various reproductive events in different regions of the female reproductive tract. PMID- 11891576 TI - Isolation and community: a review of the role of gap-junctional communication in embryonic patterning. AB - Gap junctions are specialized channels formed between the membranes of two adjacent cells. They permit the direct passage of small molecules from the cytosol of one cell to that of its neighbor, and thus form a system of cell-cell communication that exists alongside familiar secretion/receptor signaling. Gap junction states can be regulated at many levels by factors such as membrane voltage, pH, phosphorylation state, and biochemical signals. Because of the rich potential for regulation of junctional conductance, and directional and molecular gating (specificity), gap junctional communication (GJC) plays a crucial role in many aspects of normal tissue physiology, as well as in tumor progression. However, arguably the most exciting role for GJC is in the regulation of information flow that takes place during embryonic development. This review summarizes the current knowledge of how GJC controls various aspects of embryonic morphogenesis in both vertebrate and invertebrate systems. Modern molecular embryology approaches have complemented biophysical and ultrastructural data, and we are beginning to unravel the patterning roles of GJC in embryonic events such as the patterning of the embryonic left-right axis, as well as the morphogenesis of the heart and limb. Proteins from the Connexin (Cx) gene family, as well as innexins and ductin, are now beginning to be understood as the basis for GJC underlying important embryonic patterning events. PMID- 11891577 TI - Evidence that glucose-induced electrical activity in rat pancreatic beta-cells does not require KATP channel inhibition. AB - KATP-channel activity, recorded in cell-attached patches from isolated rat pancreatic beta-cells, was found to be maximally inhibited in the presence of a substimulatory concentration (5 mM) of glucose, with no further effect of higher, stimulatory glucose concentrations. KATP channel-independent effects of glucose on electrical activity were therefore investigated by incubating cells in the presence of a supramaximal concentration of tolbutamide. Addition of tolbutamide (500 mM) to cells equilibrated in the absence of glucose resulted in a rapid depolarization and electrical activity followed by a gradual repolarization and disappearence of electrical activity. Repolarization was not due to desensitization of KATP channels to the sulfonylurea, but was probably the result of activation of another K+ conductance. The subsequent application of 16 mM glucose in the continued presence of tolbutamide depolarized the cells again, leading to renewed electrical activity. Input conductance of the cells was markedly reduced by tolbutamide, reflecting KATP-channel inhibition, but was not significantly affected by the addition of glucose in the presence of the drug. In cells voltage-clamped at -70 mV, addition of glucose in the presence of tolbutamide generated a noisy inward current, probably representing activation of the volume-sensitive anion channel. KATP channel-independent activation of electrical activity by glucose was inhibited by the anion channel inhibitor 4,4' dithiocyanatostilbene-2,2'-disulphonic acid. It is concluded that the induction of electrical activity in rat pancreatic b-cells does not require inhibition of KATP channels. The KATP channel-independent mechanism could involve, at least in part, activation by glucose of the volume-sensitive anion channel. PMID- 11891578 TI - Trek-like potassium channels in rat cardiac ventricular myocytes are activated by intracellular ATP. AB - Large (111 +/- 3.0 pS) K+ channels were recorded in membrane patches from adult rat ventricular myocytes using patch-clamp techniques. The channels were not blocked by 4-AP (5 mM), intracellular TEA (5 mM) or glybenclamide (100 mM). Applying stretch to the membrane (as pipette suction) increased channel open probability (Po) in both cell-attached and isolated patches (typically, Po approximately equals 0.005 with no pressure; approximately equals 0.328 with 90 cm H2O: Vm = 40 mV, pHi = 7.2). The channels were activated by a decrease in intracellular pH; decreasing pHi to 5.5 from 7.2 increased Po to 0.16 from approx. 0.005 (no suction, Vm held at 40 mV). These properties are consistent with those demonstrated for TREK-1, a member of the recently cloned tandem pore family. We confirmed, using RT-PCR, that TREK-1 is expressed in rat ventricle, suggesting that the channel being recorded is indeed TREK-1. However, we show also that the channels are activated by millimolar concentrations of intracellular ATP. At a pH of 6 with no ATP at the intracellular membrane face, Po was 0.048 +/-0.023, whereas Po increased to 0.22 +/- 0.1 with 1 mM ATP, and to 0.348 +/- 0.13 with 3 mM (n = 5; no membrane stretch applied). The rapid time course of the response and the fact that we see the effect in isolated patches appear to preclude phosphorylation. We conclude that intracellular ATP directly activates TREK-like channels, a property not previously described. PMID- 11891579 TI - Regulation of the lemon-fruit V-ATPase by variable stoichiometry and organic acids. AB - The lemon-fruit V-ATPase can exist in two forms: nitrate-sensitive and nitrate insensitive. Here we report the results of measurements of H+ /ATP stoichiometries using two kinetic methods: one based on steady-state DpH and one based on initial rates of H+-pumping. Our findings indicate that the nitrate insensitive fruit V-ATPase has an H+ /ATP stoichiometry of ~1, while both the nitrate-sensitive fruit V-ATPase and the epicotyl V-ATPase have stoichiometries of 2, under zero-load conditions. As DpH increases, the stoichiometry of the nitrate-sensitive fruit V-ATPase decreases to 1. Under similar conditions, the stoichiometry of the epicotyl enzyme remains 2. Thus, the pH-dependent variable stoichiometry of the lemon-fruit V-ATPase may represent a key factor in juice sac vacuolar hyperacidification. On the other hand, the H+ /ATP stoichiometry of the epicotyl V-ATPase can decrease from 2 to 1 in the presence of a membrane potential. The low pH of the fruit vacuole is not due solely to the lower H+/ATP stoichiometry of its pump. We show that lumenal citrate and malate improve the coupling of both the epicotyl and fruit V-ATPases and enhance their ability to generate a pH gradient. Since citrate accumulation is restricted to fruit vacuoles, it may be another important determinant of vacuolar pH. PMID- 11891580 TI - Detection of charge movements in ion pumps by a family of styryl dyes. AB - A family of fluorescent styryl dyes was synthesized to apply them as probes that monitor the ion-translocating activity of the Na,K-ATPase and the SR Ca-ATPase, similar to the widely used dye RH421. All dyes had the same chromophore but they differed in the length of the spacer between chromophore and polar head, an isothiocyanate group, and in the lengths of the two identical acyl chains, which form the tail of the dye molecules. A number of substrate-dependent partial reactions of both P-type ATPases affected the fluorescence intensity, and the magnitude of the fluorescence changes was used to characterize the usefulness of the dyes for further application. The experimental results indicate that electrochromy is the major mechanism of these dyes. While in the case of the Na,K ATPase a single dye, 5QITC, showed larger fluorescence changes than all others, in the case of the SR Ca-ATPase all dyes tested were almost equal in their fluorescence responses. This prominent difference is interpreted as a hint that the position of the ion binding sites in both ion pumps may differ significantly despite their otherwise closely related structural features. Quench experiments with spin-labeled lipids in various positions of their fatty acids were used to gain information on the depth of the chromophore of the different dyes within the membrane dielectric, however, the spatial resolution was so poor that only qualitative information on the position of the chromophore in the lipid phase could be obtained. PMID- 11891582 TI - A ligand-dependent conformational change of the Na+/galactose cotransporter of Vibrio parahaemolyticus, monitored by tryptophan fluorescence. AB - Purification and reconstitution of the active Vibrio parahaemolyticus Na+/galactose transporter (vSGLT) enables us to do protein chemistry studies on a representative member of this class of membrane transporters. By measuring intrinsic tryptophan (Trp) fluorescence, conformational changes on the binding of substrates could be investigated. Trp fluorescence increased by 6% on the addition of saturating levels of both Na+ and D-galactose, with a K0.5 for D galactose of 0.6 mM. No change was seen on the addition of Na+ alone or by adding D-galactose in the presence of K+. The Trp fluorescence could be quenched by acrylamide, but not by Cs+or I?. In the presence of Na+ or K+ alone, of Na+ or K+ and D-galactose, of Na+ and L-glucose, or in the absence of ligands, the fluorescence quenches by acrylamide were similar. This indicated that the tryptophan exposure to acrylamide was unchanged in the presence or absence of ligands. No shifts in lem maximum were observed. To find the Trp responsible for the change in fluorescence, Trp 448 in transmembrane helix 11 in the putative sugar-binding pocket was mutated. It was found that W448F showed a similar change in Trp fluorescence upon the addition of D-galactose in the presence of Na+. We conclude that the Trp fluorescence properties of the purified and reconstituted Na+/galactose cotransporter are selectively changed by the transported substrates Na+ and D-galactose, but it is not the Trp (W448) in the sugar translocation pathway that is involved. PMID- 11891581 TI - Properties of a sodium channel (Na(x)) activated by strong depolarization of Xenopus oocytes. AB - Short (<1 sec) duration depolarization of Xenopus laevis oocytes to voltages greater than +40 mV activates a sodium-selective channel (Na(x)) with sodium permeability five to six times greater than the permeability of other monovalent cations examined, including K+, Rb+, Cs+, TMA+, and Choline+. The permeability to Li+ is about equal to that of Na+. This channel was present in all oocytes examined. The kinetics, voltage dependence and pharmacology of Na(x)distinguish it from TTX-sensitive or epithelial sodium channels. It is also different from the sodium channel of Xenopus oocytes activated by prolonged depolarization, which is more highly selective for Na+, requires prolonged depolarization to be activated, and is blocked by Li+. Intracellular Mg2+ reversibly inhibits Na(x), whereas extracellular Mg2+ does not have an inhibitory effect. Intracellular Mg2+ inhibition of Na(x), is voltage dependent, suggesting that Mg2+ binding occurs within the membrane field. Eosin is also a reversible voltage-dependent intracellular inhibitor of Na(x), suggesting that a P-type ATPase may mediate the current. An additional cytoplasmic factor is involved in maintaining Na(x) since the current runs down in internally perfused oocytes and excised membrane patches. The rundown is reversible by reintroduction of the membrane patch into oocyte cytoplasm. The cytoplasmic factor is not ATP, because ATP has no effect on Na(x) current magnitude in either cut-open or inside-out patch preparations. Extracellular Gd3+ is also an inhibitor of Na(x). Na(x) activation follows a sigmoid time course. Its half-maximal activation potential is +100 mV and the effective valence estimated from the steepness of conductance activation is 1.0. Na(x) deactivates monoexponentially upon return to the holding potential (-40 mV). The deactivation rate is voltage dependent, increasing at more negative membrane potentials. PMID- 11891583 TI - Sodium transport in triads isolated from rabbit skeletal muscle. AB - Triads and transverse tubules isolated from mammalian skeletal muscle actively accumulated Na+ in the presence of K+ and Mg-ATP. Active Na+ transport exhibited a fast single-exponential phase, lasting 2 min, followed by slower linear uptake that continued for 10 minutes. Valinomycin stimulated Na+ uptake, suggesting it decreased a pump-generated membrane potential gradient (Vm) that prevented further Na+ accumulation. At the end of the fast uptake phase transverse tubule vesicles incubated in 30 mM external [Na+] attained a ratio [Na+]in/[Na+]out=13.4. From this ratio and the transverse tubule volume of 0.35 microl/mg protein measured in this work, [Na+]in=400 mM was calculated. Determinations of active K+ transport in triads, using 86Rb+ as tracer, showed a 30% decrease in vesicular 86Rb+ content two minutes after initiating the reaction, followed by a slower uptake phase during which vesicles regained their initial 86Rb+ content after 10 minutes. Transverse tubule volume increase during active Na+ transport-as shown by light scattering changes of isolated vesicles- presumably accounted for the secondary Na+ and 86Rb+ uptake phases. These combined results indicate that isolated triads have highly sealed transverse tubules that can be polarized effectively by the Na+ pump through the generation of significant Na+ gradients. PMID- 11891584 TI - Functional expression of GFP-linked human heart sodium channel (hH1) and subcellular localization of the a subunit in HEK293 cells and dog cardiac myocytes. AB - Recent evidence suggests that biosynthesis of the human heart Na+ channel (hH1) protein is rapidly modulated by sympathetic interventions. However, data regarding the intracellular processing of hH1 in vivo are lacking. In this study we sought to establish a model that would allow us to study the subcellular localization of hH1 protein. Such a model could eventually help us to better understand the trafficking of hH1 in vivo and its potential role in cardiac conduction. We labeled the C-terminus of hH1 with the green fluorescent protein (GFP) and compared the expression of this construct (hH1-GFP) and hH1 in transfected HEK293 cells. Fusion of GFP to hH1 did not alter its electrophysiological properties. Confocal microscopy revealed that hH1-GFP was highly expressed in intracellular membrane structures. Immuno-electronmicrographs showed that transfection of hH1-GFP and hH1 induced proliferation of three types of endoplasmic reticulum (ER) membranes to accommodate the heterologously expressed proteins. Labeling with specific markers for the ER and the Golgi apparatus indicated that the intracellular channels are almost exclusively retained within the ER. Immunocytochemical labeling of the Na+ channel in dog cardiomyocytes showed strong fluorescence in the perinuclear region of the cells, a result consistent with our findings in HEK293 cells. We propose that the ER may serve as a reservoir for the cardiac Na+ channels and that the transport from the ER to the Golgi apparatus is among the rate-limiting steps for sarcolemmal expression of Na+ channels. PMID- 11891585 TI - The beta1 subunit but not the beta2 subunit colocalizes with the human heart Na+ channel (hH1) already within the endoplasmic reticulum. AB - Voltage-dependent Na+ channels are heteromultimers consisting of a pore-forming a subunit and accessory b subunits. In order to provide more insight into the trafficking and assembly of the cardiac Na+ channel complex, we investigated the subcellular localization of the Na+ channel beta1 and beta2 subunits, both in the absence and presence of the human heart Na+ channel (hH1). We fused spectrally distinct variants of the green fluorescent protein (GFP) to hH1 and to the beta1 and beta2 subunit, and expressed the optically labeled b subunits separately or in combination with hH1 in HEK293 cells. In contrast to the predominant localization of hH1 channels within the endoplasmic reticulum (ER), both beta subunits were clearly targeted to the plasma membrane when expressing their cDNAs alone. Upon coexpression of the a subunit, the beta1 subunit was efficiently retained within the ER and found to be colocalized with hH1. In contrast to this, hH1 and the beta2 subunit were not colocalized, i.e., they were detected mainly within the ER and the plasma membrane, respectively. These results indicate that hH1 and the b2 subunit are transported separately to the plasma membrane whereas the hH1/beta1 complex occurs already within the ER, which possibly facilitates trafficking of the channel complex to the plasma membrane. PMID- 11891586 TI - Stable polarized expression of hCAT-1 in an epithelial cell line. AB - Our laboratory has recently identified and cloned three cationic amino-acid transporters of human placenta. We have now examined the plasma membrane domain localization and functional expression of one of these transporters, hCAT-1, in a polarized epithelial cell line (MDCK). To facilitate identification of expressed protein we first transferred the hCAT-1 cDNA to a vector with C-terminal green fluorescent protein (GFP). The resultant hCAT-1-CT-GFP fusion protein stimulated L-[3H] lysine uptake in Xenopus oocytes. In confluent monolayers of stably transfected cells grown on porous nitrocellulose filters, saturable uptake of L [3H] lysine from the basolateral surface was stimulated 7-fold over that of untransfected cells. Concentration-dependence studies in Na+-free medium at pH 7.4 demonstrated a Km of approximately 68 +/- 13 microM and a Vmax of 970 +/-170 pmol/mg protein/min. Uptake from the apical plasma membrane surface was negligible in both transfected and untransfected cells. Consistent with these results, confocal microscopy of confluent monolayers of hCAT-1-CT-GFP-expressing cells revealed localization of the transporter solely on the basolateral domain of the cell. This is apparently the first report of a cultured polarized epithelial cell model for stable expression of a cationic amino-acid transporter. It has the potential to aid in the identification of targeting signals for transport protein localization. PMID- 11891588 TI - Regulation of the Na+/Ca2+ exchanger in rat pancreatic ducts. AB - The Ca2+ content of pancreatic juice is closely regulated by yet unknown mechanisms. One aim of the present study was to find whether rat pancreatic ducts have a Na+/Ca2+ exchanger, as found in some Ca2+ transporting epithelia. Another aim was to establish whether the exchanger is regulated by hormones/agonists affecting pancreatic secretion. Whole pancreas, pure pancreatic acini and ducts were obtained from rats and used for RT-PCR and Western blot analysis, immunohistochemistry and intracellular Ca2+ measurements using Fura-2. RT-PCR analysis indicated Na+/Ca2+-exchanger isoforms NCX1.3 and NCX1.7 in acini and pancreas. Western blot with NCX1 antibody identified bands of 70, 120 and 150 kDa in isolated ducts, acini and pancreas. Immunofluorescence experiments showed the Na+/Ca2+ exchanger on the basolateral membrane of acini and small intercalated/intralobular ducts, but in larger intralobular/extralobular ducts the exchanger was predominantly on the luminal membrane. Na+/Ca2+ exchange in ducts was monitored by changes in intracellular Ca2+ activity upon reversal of the Na+ gradient. Secretin (1 nM) and carbachol (1 mM) reduced Na+/Ca2+ exchange by 40% and 51%, respectively. Insulin (1 nM) increased Na+/Ca2+ exchange by 230% within 5 min. The present study shows that pancreatic ducts express the Na+/Ca2+ exchanger. Its distinct localization along the ductal tree and regulation by secretin, carbachol and insulin indicate that ducts might be involved in regulation of Ca2+ concentrations in pancreatic juice. PMID- 11891587 TI - Sensitivity of the plant vacuolar malate channel to pH, Ca2+ and anion-channel blockers. AB - The organic anion malate is accumulated in the central vacuole of most plant cells. Malate has several important roles in plant vacuoles, such as the maintenance of charge balance and pH regulation, as an osmolyte involved in the generation of cell turgor, and as a storage form of CO2. Transport of malate across the vacuolar membrane is important for the regulation of cytoplasmic pH and the control of cellular metabolism, particularly in plants showing crassulacean acid metabolism (CAM), in which large fluxes of malate occur during the day/night cycle. By applying the patch-clamp technique, in the whole-vacuole configuration, to isolated vacuoles from leaf mesophyll cells of the CAM plant Kalanchoe daigremontiana, we studied the regulation of the vacuolar malate channel by pH and Ca2+, as well as its sensitivity to anion-channel blockers. Malate currents were found to be insensitive to Ca2+ on the cytoplasmic side of the membrane over a range from approximately 10(-8) M to 10(-4) M. In contrast, decreasing cytoplasmic pH below 7.5 had a significant modulatory effect on channel activity, reducing malate currents by 40%, whereas increasing cytoplasmic pH above 7.5 resulted in no change in current. Several known Cl?-channel blockers inhibited the vacuolar malate currents: niflumic acid and indanoyloxyacetic acid (IAA-94) proved to be the most effective inhibitors, exerting half-maximal effects at concentrations of approximately 20 mM, suggesting that the plant vacuolar malate channel may share certain similarities with other classes of known anion channels. PMID- 11891589 TI - Ultrasound-assisted lipoplasty (UAL) in breast surgery. AB - Breast surgery has evolved significantly since the increased demand for reduced scars led to the development of minimal incision techniques. Ultrasound-assisted lipoplasty (UAL) presents important advantages when compared to traditional liposuction, such as preservation of connective structures and significant skin retraction capability. Other factors such as a favorable side-effect profile, satisfactory aesthetic results, and virtually inconspicuous scars have led us to utilize UAL in virtually all of the different breast surgery modalities carried out in our practice. Important aspects of patient selection, markings, surgical technique, and postoperative care are outlined. Ultrasonic energy is applied through superficial tunnels that lie radial to the mammary cone, with preservation of elements such as the areola, mammary ducts, and the central part of the breast's base which contains the perforators that supply the gland. Deep treatment should be applied onto adipose tissue regions and should preferably be performed in the peripheral and subcutaneous layers of the breast, with conservation of the central glandular cone to ensure maintenance of anterior projection. In selected cases, UAL is a valuable adjunct to procedures such as symmetrization, reduction mammaplasty, and breast reconstruction, permitting both volume reduction and shaping through three-dimensional retraction of connective tissue and skin. The excellent preliminary results support new indications and future developments of the technique. PMID- 11891590 TI - Three-dimensional analysis of forehead wrinkles. AB - It is important to understand the anatomical characteristics of forehead wrinkles in order to perform a forehead lift. This study aimed to elucidate the mutual relationships among tissues composing the forehead using computer software that enables the stereoscopic observation of the tissues studied in arbitrary directions. The specimens were obtained from five cadavers and prepared in serial sagittal sections. Three-dimensional images were prepared by inputting the forehead wrinkle data obtained from serial sagittal sections. Consequently, the forehead skin was found to be fixed to the superficial galea aponeurotica through fibrous septa, suggesting that movement of the frontalis muscle would be transmitted to the skin, not through the fibrous septa alone, but from the superficial galea aponeurotica closely attached to the frontalis muscle through the fibrous septa. Since the forehead muscles exhibit a stereostructure where the corrugator supercilii muscle supports three superficial forehead muscles, including the frontalis muscle, the orbicularis oculi muscle, and procerus muscle on the periosteal side, it was presumed that a sufficient effect would not be attained unless the corrugator supercilii muscle was operated on concurrently in conjunction with these muscles during a procedure involving the superficial forehead muscles in a forehead lift. Based on the findings from the three dimensional images obtained, effective tissue treatments could be achieved by performing (1) dissection between the superficial galea aponeurotica and the frontalis muscle, (2) dissection between the deep galea aponeurotica and the periosteum, and (3) a procedure incorporating the forehead muscles in order. PMID- 11891591 TI - Does liposuction influence lipidogram in females: in vivo study. AB - Suction lipectomy is one of the most commonly performed cosmetic procedures that is ideally indicated for the treatment of minimal to moderate localized fat deposits. The safety of the procedure has been addressed regarding patient selection, complications and results. Little is known about the impact of liposuction on lipid metabolism during and immediately after the procedure. Ten consecutive patients operated on for moderate volume liposuction (mean 1470 cc, range 500-2800 cc) were included in the present study. Blood samples analysis were obtained preoperatively (T1), 20 minutes after the beginning of the procedure (T2), one hour (T3), and four hours postoperatively (T4). The levels of total cholesterol, HDL cholesterol, LDL cholesterol,triglycerides, lipoprotein lipase, A1 lipoprotein, and B lipoprotein were measured from T1 to T4. All the values were increased at T2 and T3, but remained under upper normal values and nearly complete return to baseline was observed at T4, underlining the safety of the procedure on a metabolic basis. PMID- 11891592 TI - Severe abdominal wall necrosis after ultrasound-assisted liposuction. AB - The complicated case of a 44-year-old white female following ultrasound-assisted liposuction of the entire abdomen is presented. In this case the postoperative course was complicated by hematoma, treated conservatively. During following weeks extensive cutaneous necrosis over the abdomen developed. After four weeks the patient presented to our institution with purulent discharge. After surgical revision, requiring excision of the abdominal wall necrosis, a significant residual abdominal wall defect remained. After three further revisions, removal of residual necrotic fat, irrigation, and temporary coverage with a synthetic dressing, infection cleared. At this point, split thickness skin grafting was possible. The healing in of the skin grafts was complete, eventually enabling wound closure and successful reconstruction of the abdominal wall. PMID- 11891593 TI - A new concept in male chest reshaping: anatomical pectoral implants and liposculpture. AB - A new concept of male chest reshaping has been developed. In a population of 12 patients, 10 had a male chest enhancement using only a new shaped silicon gel implant, and two had a combination of liposuction and pectoral implants. The very natural appearance of this new implant, with or without a liposuction, seems to dramatically improve the quality of the cosmetic results. PMID- 11891594 TI - Zygomaticus major advancement as an adjunct to lower blepharoplasty. AB - Sagging of the mid-face, deep nasolabial folds, and flat malar eminence often compromise the rejuvenation effect produced by upper and lower blepharoplasty. A simple surgical technique based on advancement of the zygomaticus major origin was developed as an adjunct to lower blepharoplasty to address this problem. The technique is predominantly suitable for ambulatory surgery setup and can be easily performed under local anesthesia and sedation. It offers rejuvenation of the midface and correction of the nasolabial fold. PMID- 11891595 TI - A new technique for levator lengthening to treat upper eyelid retraction: the orbital septal flap. AB - Correction of upper eyelid retraction can be achieved by numerous techniques. We have developed a new flap, the orbital septal flap, to interpose between the recessed levator complex and the tarsus to correct the retracted upper eyelid of a young girl. The orbital septum is a facial structure; it is readily available and easy to dissect. The flap acts like a vascularited spacer without the problem of resorption; normal anatomical continuity of the levator mechanism can be functionally restored. We believe the orbital septal flap is a promising technique for correcting upper eyelid retraction; however, more case studies are needed. PMID- 11891596 TI - Correction of bone deformity after resection of dermoid cyst using artificial dermis implantation. AB - We report cases of the dermoid cyst patients with a bone deformity that were treated with artificial dermis implantation. After resecting the tumor, three or five sheets of artificial dermis (Terudermis, Terumo Co. Ltd, Tokyo, Japan) are placed into the depressed site in order to create symmetrical surface with the contralateral side. Neither of the cases presented here showed allergic or infectious reaction during postoperative follow-up period. In both cases, the texture of the implanted region is almost the same as the contralateral side and is almost symmetrical. We believe that artificial dermis may be a useful implantation material to correct depressions in the patient with dermoid cysts. PMID- 11891597 TI - Multidisciplinary and multistage treatment of complex facial trauma: case report. AB - The improved performance of modern automobiles often results in higher driving speeds, rendering traffic accidents much more devastating. Many of our facially injured patients require multidisciplinary and multistage treatment, which is often both sophisticated and radical. We present a patient with complex facial trauma, discuss his injury characteristics and the multistage treatment performed, and review the literature. PMID- 11891598 TI - Breast augmentation as an incentive in recovering from anorexia. AB - In a survey carried out on 229 subjects who had undergone an augmentative mammaplasty it was possible to verify a postoperative increase in weight in 25 cases, four of which were clearly anorexic. We hypothesized that a change in perception of one's body proportions after the insertion of implants, might have been a determinant in blocking the mechanism leading to anorexia, or at least in the continuation of the recovering process. The aim of this article is obviously not to state that augmentative mammaplasty can be a kind of therapy for anorexia. Instead, we want to underline how a more pleasant contour of some body areas can have a role in solving deeper psychological problems. PMID- 11891599 TI - Tuberous breast: a surgical challenge. AB - Since the presentation of the tuberous breast deformity by Rees and Aston in 1976, many surgical procedures have been developed, but the correction of such a deformity still remains a surgical challenge. The authors report the last cases treated in the Department of Plastic and Reconstructive Surgery of Sao Joao Hospital-Porto and discuss about the ideal procedure which should be used according to the type of deformity. They emphasize the periareolar approach and the good results obtained by the Liacyr Ribeiro technique. PMID- 11891600 TI - Suspicious node found at the time of reduction mammaplasty. AB - A case of a patient with a suspicious glandular node found during reduction mammaplasty is described. The preoperative search for such nodes, the management of cases on which a suspicious node is found intraoperatively, and a situation on which the diagnosis of breast cancer is made during histology are discussed. When a suspicious small node (with a diameter up to 2 cm) is detected during a cosmetic breast surgery, lumpectomy can be performed. It may be a definite surgical treatment, depending on stage and tumor type. In the case presented, histology revealed intraductal papilloma, a benign tumor, therefore lumpectomy was a suitable procedure with an acceptable cosmetic result. With the increased incidence of breast cancer, this situation will happen more often and technical options for the management of such cases deserves the attention of plastic surgeons. PMID- 11891601 TI - Surgical treatment of rhinophyma: a comparison of techniques. AB - Rhinophyma is a rare disease that primarily affects Caucasian men in the fifth to seventh decades of life, characterized by a progressive thickening of nasal skin, which produces a disfiguring soft-tissue hypertrophy of the nose. Severe cosmetic deformity and impairment of breathing may coexist, making the surgical treatment necessary. The authors are conscious that in literature there is not agreement about the ideal treatment of rhinophyma, nevertheless they wish to give their contribution according to their experience with different treatment modalities such as the scalpel, the electrocautery, the dermabrader, and the carbon dioxide laser. The authors believe the scalpel, in association with bipolar electrocautery and local infiltration of dilute epinephrine to reduce bleeding, is the safest means to preserve the underlying sebaceous gland fundi and permit a spontaneous re-epithelialization scarring-free. PMID- 11891602 TI - Case report of phototherapy for a clinical port wine lesion. AB - Port wine lesions have been a difficult problem for plastic surgeons to treat effectively. This article will offer a new way to treat the port wine lesion with intense pulse light, and will give an example of a very difficult port wine lesion and the results that this particular method can achieve. A 39-year-old, with a lesion along the V1, V2 trigeminal distribution, was treated with eight treatments over a span of ten months. The patient was never treated before for this lesion and now desired treatment due to nodularity and bleeding. This patient had a very good response to therapy and had no side effects. He had minimal difficulty tolerating the treatment as time went on, although he originally needed local anesthetic for pain control around the treatment area. PMID- 11891603 TI - Proportionality in Asian and North American Caucasian faces using neoclassical facial canons as criteria. AB - Nine projective linear measurements were taken to determine morphometric differences of the face among healthy young adult Chinese, Vietnamese, and Thais (60 in each group) and to assess the validity of six neoclassical facial canons in these populations. In addition, the findings in the Asian ethnic groups were compared to the data of 60 North American Caucasians. The canons served as criteria for determining the differences between the Asians and Caucasians. In neither Asian nor Caucasian subjects were the three sections of the facial profile equal. The validity of the five other facial canons was more frequent in Caucasians (range: 16.7-36.7%) than in Asians (range: 1.7-26.7%). Horizontal measurement results were significantly greater in the faces of the Asians (en-en, al-al, zy-zy) than in their white counterparts; as a result, the variation between the classical proportions and the actual measurements was significantly higher among Asians (range: 90-100%) than Caucasians (range: 13.3-48%). The dominant characteristics of the Asian face were a wider intercanthal distance in relation to a shorter palpebral fissure, a much wider soft nose within wide facial contours, a smaller mouth width, and a lower face smaller than the forehead height. In the absence of valid anthropometric norms of craniofacial measurements and proportion indices, our results, based on quantitative analysis of the main vertical and horizontal measurements of the face, offers surgeons guidance in judging the faces of Asian patients in preparation for corrective surgery. PMID- 11891604 TI - Induction and recovery phases of methacholine-induced bronchoconstriction using FEV1 according to the degree of bronchial hyperresponsiveness. AB - Forty-eight patients suffering from intermittent bronchial asthma underwent methacholine challenge test. Response was stronger in 29 patients and less pronounced in 19. The two groups had the same characteristics except for the cumulative methacholine dose which was lower in severe hyperresponsiveness. The patients were studied both in the phase of induced bronchospasm and in the subsequent phase of spontaneous recovery. Dose-response curves to methacholine were analyzed as FEV1% decline/methacholine dose for the induction phase of bronchoconstriction and as FEV1% increase*methacholine dose/time after PD20FEV1 for the recovery phase. The phase of induced bronchospasm as well as spontaneous recovery had a linear pattern in severe hyperresponsiveness; in patients with moderate response, induced bronchoconstriction had a curvilinear pattern whereas spontaneous recovery had a linear pattern. This latter group had to break down an amount of methacholine that was fivefold greater than the former, therefore the mechanism of local homeostasis recovery may be more efficient in moderate hyperresponsiveness. However, in both groups recovery after the bronchospasm was not complete after 60 min (p < 0.01 versus baseline). Furthermore, recovery was faster in the first 15 min than in the remaining time. In conclusion the behavior of methacholine-induced bronchospasm and its spontaneous recovery in both severe and moderate hyperresponsiveness seem to be different although several and not well-established mechanisms may be responsible for this phenomenon. PMID- 11891605 TI - Kv channel subunit expression in rat pulmonary arteries. AB - Hypoxic pulmonary vasoconstriction (HPV) is a mechanism whereby capillary perfusion is modulated to match alveolar ventilation by diverting blood flow away from poorly ventilated regions of the lung. K+ channels, sensitive to changes in oxygen tension, are thought to play a pivotal role in initiating contraction of pulmonary arterial smooth muscle cells. However, the specific channel subtypes involved have not yet been identified. Using RT-PCR, we have investigated the expression of delayed rectifying (Kv) channel mRNA in rat main and small pulmonary arteries and, for comparison, the systemic mesenteric artery. We have identified and fully sequenced a rat Kv9.2 cDNA and also demonstrated the presence of Kv1.7 and Kv4.1. The presence and relative distribution of Kv1.2, Kv1.5, Kv2.1, and Kv9 mRNA is consistent with the proposed contribution of these subunits to oxygen sensing by K channels, previously described in pulmonary arteries. Our data addresses the controversy relating to the likely distribution of Kv channels involved in oxygen sensing without necessarily implying that such subunits are directly responsible for this process. The differential expression of other subunits, particularly Kv4, indicates that these too may have a role in HPV, revealing the need for further biophysical evaluation of these channel subtypes. PMID- 11891606 TI - Different expression of endothelin in the bronchoalveolar lavage in patients with pulmonary diseases. AB - Endothelin (ET) is a broncho- and vasoconstrictive cytokine, but it also possesses proinflammatory and mitogenic activity. It is suggested to be involved in the pathogenesis of fibrotic lung diseases. We analyzed the concentration of ET 1 in the bronchoalveolar lavage (BAL) fluid in 95 patients with different lung diseases, among them 41 patients with interstitial lung diseases (13 fibrosing alveolitis in systemic sclerosis (FASS), 9 idiopathic pulmonary fibrosis (IFP), 8 sarcoidosis (S), 6 occupational lung disease (OLD), 5 other alveolitidies A), 27 patients with pneumonia, and 8 patients with chronic obstructive pulmonary disease (COPD). A heterogeneous group of 19 patients served as controls. The median ET concentration was 3.3 pg/ml. Significantly higher concentration was found in patients with FASS (5.8 pg/ml), IPF (5.0 pg/ml), and S (5.1 pg/ml) compared with OLD (2.8 pg/ml), A (1.9 pg/ml), COPD (1.5 pg/ml), and the control group (2.5 pg/ml). In pneumonia, the elevated ET concentration (4.1 pg/ml) was accompanied by a high alveolocapillary leakage. When normalized to BAL albumin concentration, only FASS presented with significantly elevated ET/albumin in the BAL compared with the control group (134.5 vs. 56.l pg/mg, p < 0.05). There were no correlations between ET and BAL differential cell count or pulmonary function tests. In current smokers, ET in BALF was significantly higher compared with non- or ex-smokers (3.9 vs. 2.0 pg/ml, p < 0.01), but not so the ET/albumin ratio (65.0 vs. 62.5 pg/mg). In summary, ET in the BAL is differentially expressed in distinct inflammatory and interstitial lung disease. Consistently high concentrations are found in FASS and elevated ET concentration could be discussed in IPF, sarcoidosis, and pneumonia. ET concentration in BAL is influenced by current smoking habits. PMID- 11891607 TI - Gamma-interferon and soluble interleukin 2 receptor in tuberculous pleural effusion. AB - To analysis the difference between systemic and local pleural T cell response in pulmonary tuberculosis, we analyzed interferon (IFN)-gamma and soluble interleukin-2 receptor (sIL-2R) in peripheral blood mononuclear cells (PBMC) culture supernatants and in pleural effusion (PE). We also investigated the association of pleural INF-gamma and sIL-2R levels with development of residual pleural thickening (RPT). The subjects in this study included patients with active pulmonary tuberculosis with or without PE (n = 46), those with nontuberculous PE (n = 32), and healthy tuberculin reactors (n = 20). Measurement of IFN-gamma and sIL-2R were made by ELISA. In pulmonary tuberculosis, IFN-gamma and sIL-2R concentrations in PBMC culture supernatants were lower than those of healthy tuberculin reactors (IFN-gamma; 258.4 +/-111.5 pg/mL versus 2792.5 +/ 633.2 pg/mL, sIL-2R; 1465.0 +/-144.4 pg/mL versus 4777.1 +/-178.5 pg/mL, p < 0.05), whereas IFN-gamma and sIL-2R concentrations in PE were higher than those from nontuberculous pleural effusion (IFN-gamma; 1154.4 +/-252.4 pg/mL versus 292.0 +/-68.9 pg/mL, sIL-2R; 9805.2 +/-978.9 pg/mL versus 3426.7 +/-695.6 g/mL, p < 0.05). IFN-gamma and sIL-2R in PBMC culture supernatants were significantly lower in tuberculat patients with PE than those without PE, and the patients with a high value of IFN-gamma or sIL-2R in PE showed a low value of IFN-gamma or sIL 2R in PBMC culture supernatant, respectively. Patients with RPT had significantly higher IFN-gamma and sIL-2R values in their PE compared with those without RPT. These findings suggest that diminished systemic Th1 response in tuberculosis results from the accumulation of activated Th1 cell to the disease site, and that levels of IFN-gamma and sIL-2R in PE are useful posttreatment markers of RPT. PMID- 11891608 TI - Effect of recombinant human DNase on alpha1-proteinase inhibitor function: an experimental approach to the combined clinical use of rhDNase and alpha1-PI in CF patients. AB - Chronic inflammation in cystic fibrosis (CF) airways leads to high concentrations of deoxyribonucleic acid (DNA) and neutrophil elastase (NE). Both play a major role in CF lung pathophysiology and are aims of new therapeutic approaches: rhDNase degrades highly viscosic DNA and alpha1-proteinase inhibitor (alpha1-PI) inhibits NE activity and thereby pulmonary inflammation and hypersecretion. Given the reports on increased sputum NE concentrations upon rhDNase inhalation, there is a rationale for a combined rhDNase/alpha1-PI treatment. With the question of whether a combined therapy is feasible, we first investigated in vitro whether incubation of CF sputum with rhDNase changes proteolytic and secretagogue activity of sputum supernatants and its inhibition by alpha1-PI. Next, we studied whether incubation of alpha1-PI with rhDNase impairs the inhibitory effect of alpha1-PI on proteolytic activity of NE and the inhibitory effect of alpha1-PI on NE-induced secretion from a human mucoepidermoid cell line. Incubation of CF sputum with rhDNase led to a twofold increase in sputum NE activity. Correspondingly, the inhibitory effect of alpha1-PI on sputum NE activity and on secretion induced by these sputum samples was significantly reduced by rhDNase. Preincubation of alpha1-PI with rhDNase significantly reduced the inhibitory effect of alpha1-PI on purified NE activity and on NE-induced secretion. However, this effect was limited to alpha1-PI concentrations lower than those achievable after inhalation. Therefore, impairment of alpha1-PI function by rhDNase is not likely to be relevant in vivo, provided that a sufficient dosage of alpha1-PI is inhaled. PMID- 11891611 TI - Technetium-99m tetrofosmin chest imaging related to p-glycoprotein expression for predicting the response with paclitaxel-based chemotherapy for non-small cell lung cancer. AB - Our aim was to use technetium-99m tetrofosmin (Tc-TF) uptake in non-small cell lung cancer (NSCLC) for predicting the chemotherapeutic response of NSCLC to paclitaxel and to compare the results with the expression of multidrug resistance (MDR) - P-glycoprotein (Pgp). Twenty patients with advanced NSCLC were enrolled in this study before chemotherapy with paclitaxel. Tc-TF chest imaging was performed to calculate early and delayed tumor-to-normal lung (T/NL) count density ratios, as well as washout indexes (WIs). Immunohistochemical analyses were performed on multiple nonconsecutive sections of the biopsy specimens to detect Pgp expression. The response to chemotherapy was evaluated by clinical and radiological methods in the third month after completion of treatment. The early and delayed T/NL count-density ratios of patients with good response were significantly higher than those of patients with poor response (p <0.05). However, no significant difference in WI between the two groups of patients was found (p > 0.05). A significantly higher incidence of good response was found in patients with negative Pgp expression (100%) than in patients with positive Pgp expression (40%) (p <0.05). Significantly higher early and delayed T/NL count density ratios as well as decreased WIs were found in patients with negative Pgp expression than in patients with positive Pgp expression. However, other prognostic factors (age, sex, body weight loss, performance status, tumor stage, and tumor cell type) were not significantly different between the patients with good response and those with poor response. Because Tc-TF chest images can correctly represent the expression of Pgp in NSCLC, it can accurately predict the chemotherapeutic response to paclitaxel. PMID- 11891612 TI - The diaphragm and dyspnea during chemically stimulated breathing in a subset of patients with diabetes. AB - In patients with insulin-dependent diabetes mellitus (IDDM) isolated peripheral airway involvement may give rise to inspiratory threshold load (ITL) contributing to dyspnea. Based on the reported evidence of a greater increase in end expiratory lung volume (EELV) with hypoxia than with hypercapnia in IDDM, we wondered whether, and to what extent in the two conditions, EELV contribute to perception of dyspnea (PD). We studied five nonsmokers aged between 19 and 45, with IDDM under good metabolic control and five normal control subjects matched for age. In each patient, we evaluated the electromyographic activity of the diaphragm (Edi), the swings of esophageal (Pessw), gastric (Pgsw), and transdiaphragmatic (Pdisw = Pgsw-Pessw) pressures; PD was assessed by a modified Borg scale during hypercapnic-hyperoxic (HCH) and hypoxic-isocapnic (HIC) stimulation. Change in inspiratory capacity (IC) was considered the mirror image of increase in EELV, that is, dynamic hyperinflation (DH), while intrinsic positive end inspiratory pressure (PEEPi) was measured as an index of inspiratory threshold load (ITL). In controls, Edi and Pdi but not their ratio (Edi/Pdi) related to Borg. In patients the following was found: (1) with each of the two stimuli, for any given Edi, Pdi, and Edi/Pdi ratio, there was greater Borg than in controls, (2) a similar increase in ITL and DH with HCH and HIC, (3) Edi/Pdi related to Borg similarly with HCH as with HIC. In conclusion, in controls, Edi and Pdi were associated with the perception of dyspnea similarly with the two chemical stimuli. In this subset of patients with IDDM, Edi/Pdi ratio throughout increase in EELV and ITL was found to affect the perception of dyspnea in hypoxia to a similar extent as in hypercapnia. PMID- 11891613 TI - In utero and postnatal exposure of Wistar rats to low frequency/high intensity noise depletes the tracheal epithelium of ciliated cells. AB - Chronic exposure of men or rodents to low frequency/high intensity (LFHI) noise causes a number of systemic changes that make up the so-called vibroacoustic disease (VAD), a disorder that includes alterations of the respiratory system, namely, of its epithelial layer. We have investigated here the susceptibility of the tracheal epithelium of Wistar rats to in utero and postnatal exposure to LFHI noise by comparing its ultrastructure with that of the tracheal epithelium of control rats and of animals exposed to LFHI noise only after reaching adulthood (8 weeks of age). Scanning electron microscopy (SEM) of the inner surface of rat trachea was used to determine the relative areas covered by ciliated and non ciliated cells. In rats that were exposed in utero and postnatally to LFHI noise, we observed that out of 100 microm(2) of tracheal epithelium only 31 +/- 14 microm(2) were covered by cilia, whereas in control rats; ciliated cells occupied an average of 60 +/- 18 microm(2) out of 100 microm(2) of the epithelium; this difference between the two groups was statistically significant (p <0.05). In rats that were exposed to LFHI noise only after reaching adulthood, cilia covered 55 +/- 22 microm(2) out of 100 microm(2) of the luminal surface of the trachea, a value that, although lower than that of controls, was not found to be statistically different. We conclude that (1) the tracheal ciliated cells are damaged by exposure of rats to LFHI noise if the animals are kept under this environmental aggression during in utero and postnatal periods; (2) tracheal ciliated cells from adult rats are more resistant to the deleterious effects of LFHI noise than pleura or lung alveolar cells that were shown before to undergo marked changes upon chronic exposure of rats to LFHI noise. These findings suggest a note of caution regarding pregnant women and young children: they should be prevented from areas where LFHI noise occurs, namely, in aircraft and textile industries where this type of environmental hazard is often present. PMID- 11891614 TI - High levels of nitric oxide in individuals with pulmonary hypertension receiving epoprostenol therapy. AB - Lack of vasodilator substances, such as nitric oxide (NO), has been implicated in the development of pulmonary hypertension, but the pathogenesis of the disease remains speculative. We hypothesized that NO plays a role in the pathogenesis of primary pulmonary hypertension (PPH), and may serve as a sensitive and specific marker of disease progression and/or severity. To test this, exhaled NO and pulmonary artery pressure were measured in individuals with PPH and secondary pulmonary hypertension (SPH) on various therapies, including the potent vasodilator epoprostenol (prostacyclin), compared with healthy controls. NO in exhaled breath of individuals with PPH was lower than SPH or control (p<0.05). In contrast, exhaled NO of individuals with PPH or SPH receiving epoprostenol was strikingly higher than PPH or SPH individuals not receiving epoprostenol, or controls. Concomitant with higher NO levels, right ventricular systolic pressure of individuals significantly decreased with epoprostenol. Importantly, in paired measures of exhaled NO before and after epoprostenol, NO increased in all pulmonary hypertensive individuals 24 h after initiation of epoprostenol therapy (p<0.05). NO may be a useful noninvasive marker of pulmonary hypertension severity and response to prostacyclin therapy. PMID- 11891615 TI - Effect of ventilation mode on gas exchange during partial liquid ventilation at different perfluorocarbon doses in surfactant-depleted lung. AB - Although gas ventilation is an integral part of partial liquid ventilation (PLV), the role of ventilation mode during PLV is not established, especially at a varying perfluorocarbon dose. In 10 surfactant-depleted rabbits, PLV was performed at a low dose (10 ml/kg) and at a functional residual capacity (FRC) dose (30 ml/kg) of perfluorodecalin in pressure-control (PC) and volume-control (VC) modes in balanced sequence. In these four PLV trials, PC mode was adjusted to be identical to VC mode with regard to tidal volume and inspiratory-to expiratory ratio. PaO2 during PLV in PC mode was higher than in VC mode at the Low dose (159 plus minus 93 mm Hg, 115 plus minus 75 mm Hg, respectively: p = 0.005) and at the FRC dose (228 +/- 114 mm Hg, 164 +/- 104 mm Hg, respectively: p = 0.002). PaCO2 during PLV in PC mode was lower than in VC mode at the Low dose (59 +/- 18 mm Hg, 72 +/- 20 mm Hg, respectively: p = 0.005), whereas PaCO2 at the FRC dose was not different between modes. Curves of inspiratory flow appeared least deformed with PLV in PC mode at the Low dose, whereas they were saw-tooth deformed with PLV in VC mode at both doses. Actual time for inspiratory gas flow during PLV was shorter in PC mode compared with VC mode at both doses. In conclusion, in surfactant-depleted rabbit, gas exchange during PLV was better with PC mode compared with VC mode, especially at a low perfluorocarbon dose. Given the same tidal volume, PC appeared to insufflate the perfluorocarbon-filled lung better than VC at both low and FRC doses of perfluorocarbon. PMID- 11891617 TI - Genome scans provide evidence for low-HDL-C loci on chromosomes 8q23, 16q24.1 24.2, and 20q13.11 in Finnish families. AB - We performed a genomewide scan for genes that predispose to low serum HDL cholesterol (HDL-C) in 25 well-defined Finnish families that were ascertained for familial low HDL-C and premature coronary heart disease. The potential loci for low HDL-C that were identified initially were tested in an independent sample group of 29 Finnish families that were ascertained for familial combined hyperlipidemia (FCHL), expressing low HDL-C as one component trait. The data from the previous genome scan were also reanalyzed for this trait. We found evidence for linkage between the low-HDL-C trait and three loci, in a pooled data analysis of families with low HDL-C and FCHL. The strongest statistical evidence was obtained at a locus on chromosome 8q23, with a two-point LOD score of 4.7 under a recessive mode of inheritance and a multipoint LOD score of 3.3. Evidence for linkage also emerged for loci on chromosomes 16q24.1-24.2 and 20q13.11, the latter representing a recently characterized region for type 2 diabetes. Besides these three loci, loci on chromosomes 2p and 3p showed linkage in the families with low HDL-C and a locus on 2ptel in the families with FCHL. PMID- 11891618 TI - Geographic and haplotype structure of candidate type 2 diabetes susceptibility variants at the calpain-10 locus. AB - Recently, a positional cloning study proposed that haplotypes at the calpain-10 locus (CAPN10) are associated with increased risk of type 2 diabetes, or non insulin-dependent diabetes mellitus, in Mexican Americans, Finns, and Germans. To inform the interpretation of the original mapping results and to look for evidence for the action of natural selection on CAPN10, we undertook a population based genotyping survey of the candidate susceptibility variants. First, we genotyped sites 43, 19, and 63 (the haplotype-defining variants previously proposed) and four closely linked SNPs, in 561 individuals from 11 populations from five continents, and we examined the linkage disequilibrium among them. We then examined the ancestral state of these sites by sequencing orthologous portions of CAPN10 in chimpanzee and orangutan (the identity of sites 43 and 19 was further investigated in a limited sample of other great apes and Old World and New World monkeys). Our survey suggests larger-than-expected differences in the distribution of CAPN10 susceptibility variants between African and non African populations, with common, derived haplotypes in European and Asian samples (including one of two proposed risk haplotypes) being rare or absent in African samples. These results suggest a history of positive natural selection at the locus, resulting in significant geographic differences in polymorphism frequencies. The relationship of these differences to disease risk is discussed. PMID- 11891619 TI - Undergraduate research, education and the future of science. PMID- 11891620 TI - Undergraduate research experiences: the translation of science education from reading to doing. PMID- 11891621 TI - Using web-based animations to teach histology. AB - We have been experimenting with the use of animations to teach histology as part of an interactive multimedia program we are developing to replace the traditional lecture/laboratory-based histology course in our medical and dental curricula. This program, called HistoQuest, uses animations to illustrate basic histologic principles, explain dynamic processes, integrate histologic structure with physiological function, and assist students in forming mental models with which to organize and integrate new information into their learning. With this article, we first briefly discuss the theory of mental modeling, principles of visual presentation, and how mental modeling and visual presentation can be integrated to create effective animations. We then discuss the major Web-based animation technologies that are currently available and their suitability for different visual styles and navigational structures. Finally, we describe the process we use to produce animations for our program. The approach described in this study can be used by other developers to create animations for delivery over the Internet for the teaching of histology. PMID- 11891622 TI - The human cadaver in the age of biomedical informatics. AB - Major national and international critiques of the medical curriculum in the 1980s noted the following significant flaws: (1) over-reliance on learning by rote memory, (2) insufficient exercise in analysis and synthesis/conceptualization, and (3) failure to connect the basic and clinical aspects of training. It was argued that the invention of computers and related imaging techniques called to question the traditional instruction based on the faculty-centered didactic lecture. In the ensuing reform, which adopted case-based, small group, problem based learning, time allotted to anatomical instruction was severely truncated. Many programs replaced dissection with prosections and computer-based learning. We argue that cadaver dissection is still necessary for (1) establishing the primacy of the patient, (2) apprehension of the multidimensional body, (3) touch mediated perception of the cadaver/patient, (4) anatomical variability, (5) learning the basic language of medicine, (6) competence in diagnostic imaging, (7) cadaver/patient-centered computer-assisted learning, (8) peer group learning, (9) training for the medical specialties. Cadaver-based anatomical education is a prerequisite of optimal training for the use of biomedical informatics. When connected to dissection, medical informatics can expedite and enhance preparation for a patient-based medical profession. Actual dissection is equally necessary for acquisition of scientific skills and for a communicative, moral, ethical, and humanistic approach to patient care. Anat Rec (New Anat) 269:20-32, 2002. PMID- 11891624 TI - Morphology-based systematics (MBS) and problems with fossil hominoid and hominid systematics. AB - The generalized/primitive nature of the hominoid dentition and often fragmentary nature of fossils, coupled with enthusiastic optimism for making revolutionary finds, has wreaked havoc with recognition of early human ancestors and reconstruction of fossil hominoid phylogeny. As such, the history of paleoanthropology is one of repeated misidentification of fossil ancestors and of occasional fraud. Although this history has led many workers to lose confidence in morphology based systematics (MBS), past and present misidentifications are actually due to a disregard of systematic methodology. Systematics depends on the continuity of life and gains its objectivity largely from the order alpha taxonomy imposes on morphologic discontinuities in closely related taxa (i.e., species and genera). Transformation of characters fixed in species into character complexes, as manifested in taxa nested at different levels of relationship, form the foundation for higher-level taxonomy and for phylogeny. Because in most cases, hominoid fossils are unable to provide the data needed to resolve alpha taxonomy, classification and phylogeny of fossil taxa must be guided by analogies to living taxa. Hominid and hominoid fossil taxonomy and phylogeny, however, has been based largely on preevolutionary notions and on misinterpretations of the polarity of assumed diagnostic characters. More often than not, fossils lack resolution for the taxonomic level or rank they are assigned to and taxa are erected without appropriate analogies to living forms. As such, phylogenies based on these classifications are unlikely to be correct. More in-depth anatomical studies that are in accordance with systematic methodology are likely to hold the key to correctly classifying fossils and unraveling hominoid and hominid phylogeny. PMID- 11891623 TI - Neural regeneration and the peripheral olfactory system. AB - The peripheral olfactory system is able to recover after injury, i.e., the olfactory epithelium reconstitutes, the olfactory nerve regenerates, and the olfactory bulb is reinnervated, with a facility that is unique within the mammalian nervous system. Cell renewal in the epithelium is directed to replace neurons when they die in normal animals and does so at an accelerated pace after damage to the olfactory nerve. Neurogenesis persists because neuron-competent progenitor cells, including transit amplifying and immediate neuronal precursors, are maintained within the population of globose basal cells. Notwithstanding events in the neuron-depleted epithelium, the death of both non-neuronal cells and neurons directs multipotent globose basal cell progenitors, to give rise individually to sustentacular cells and horizontal basal cells as well as neurons. Multiple growth factors, including TGF-alpha, FGF2, BMPs, and TGF-betas, are likely to be central in regulating choice points in epitheliopoiesis. Reinnervation of the bulb is rapid and robust. When the nerve is left undisturbed, i.e., by lesioning the epithelium directly, the projection of the reconstituted epithelium onto the bulb is restored to near-normal with respect to rhinotopy and in the targeting of odorant receptor-defined neuronal classes to small clusters of glomeruli in the bulb. However, at its ultimate level, i.e., the convergence of axons expressing the same odorant receptor onto one or a few glomeruli, specificity is not restored unless a substantial number of fibers of the same type are spared. Rather, odorant receptor-defined subclasses of neurons innervate an excessive number of glomeruli in the rough vicinity of their original glomerular targets. PMID- 11891625 TI - Computational studies on nucleic acid building blocks. PMID- 11891626 TI - Electronic properties, hydrogen bonding, stacking, and cation binding of DNA and RNA bases. AB - This review summarizes results concerning molecular interactions of nucleic acid bases as revealed by advanced ab initio quantum chemical (QM) calculations published in last few years. We first explain advantages and limitations of modern QM calculations of nucleobases and provide a brief history of this still rather new field. Then we provide an overview of key electronic properties of standard and selected modified nucleobases, such as their charge distributions, dipole moments, polarizabilities, proton affinities, tautomeric equilibria, and amino group hybridization. Then we continue with hydrogen bonding of nucleobases, by analyzing energetics of standard base pairs, mismatched base pairs, thio-base pairs, and others. After this, the nature of aromatic stacking interactions is explained. Also, nonclassical interactions in nucleic acids such as interstrand bifurcated hydrogen bonds, interstrand close amino group contacts, C [bond] H...O interbase contacts, sugar-base stacking, intrinsically nonplanar base pairs, out of-plane hydrogen bonds, and amino-acceptor interactions are commented on. Finally, we overview recent calculations on interactions between nucleic acid bases and metal cations. These studies deal with effects of cation binding on the strength of base pairs, analysis of specific differences among cations, such as the difference between zinc and magnesium, the influence of metalation on protonation and tautomeric equlibria of bases, and cation-pi interactions involving nucleobases. In this review, we do not provide methodological details, as these can be found in our preceding reviews. The interrelation between advanced QM approaches and classical molecular dynamics simulations is briefly discussed. PMID- 11891627 TI - Beyond nucleic acid base pairs: from triads to heptads. AB - Hydrogen-bonded base pairs are an important determinant of nucleic acid structure and function. However, other interactions such as base-base stacking, base backbone, and backbone-backbone interactions as well as effects exerted by the solvent and by metal or NH(4)(+) ions also have to be taken into account. In addition, hydrogen-bonded base complexes involving more than two bases can occur. With the rapidly increasing number and structural diversity of nucleic acid structures known at atomic detail higher-order hydrogen-bonded base complexes, base polyads, have attracted much interest. This review provides an overview on the occurrence of base polyads in nucleic acid structures and describes computational studies on these nucleic acid building blocks. PMID- 11891628 TI - A theoretical investigation on the effect of remote amino groups in hydrogen bonding of nucleic acids. AB - The effect of remote amino groups in the H-bonding of complementary bases of duplex and triplex DNA has been investigated in the gas phase by means of density functional theory. It is found that amino groups incorporated in regions of the purine that are distant from the H-bonding sites might have a notable influence on the stability of H-bonded dimers. The results show that, in addition to primary and secondary interactions, polarization effects can be relevant for the determination of hydrogen bonding in nucleobases. PMID- 11891629 TI - Ab initio conformational analysis of nucleic acid components: intrinsic energetic contributions to nucleic acid structure and dynamics. AB - In recent years, the use of high-level ab initio calculations has allowed for the intrinsic conformational properties of nucleic acid building blocks to be revisited. This has provided new insights into the intrinsic conformational energetics of these compounds and its relationship to nucleic acids structure and dynamics. In this article we review recent developments and present new results. New data include comparison of various levels of theory on conformational properties of nucleic acid building blocks, calculations on the abasic sugar, known to occur in vivo in DNA, on the TA conformation of DNA observed in the complex with the TATA box binding protein, and on inosine. Tests of the Hartree Fock (HF), second-order Moller-Plesset (MP2), and Density Functional Theory/Becke3, Lee, Yang and Par (DFT/B3LYP) levels of theory show the overall shape of backbone torsional energy profiles (for gamma, epsilon, and chi) to be similar for the different levels, though some systematic differences are identified between the MP2 and DFT/B3LYP profiles. The east pseudorotation energy barrier in deoxyribonucleosides is also sensitive to the level of theory, with the HF and DFT/B3LYP east barriers being significantly lower (approximately 2.5 kcal/mol) than the MP2 counterpart (approximately 4.0 kcal/mol). Additional calculations at various levels of theory suggest that the east barrier in deoxyribonucleosides is between 3.0 and 4.0 kcal/mol. In the abasic sugar, the west pseudorotation energy barrier is found to be slightly lower than the east barrier and the south pucker is favored more than in standard nucleosides. Results on the TA conformation suggest that, at the nucleoside level, this conformation is significantly destabilized relative to the global energy minimum, or relative to the A- and B-DNA conformations. Deoxyribocytosine would destabilize the TA conformation more than other bases relative to the A-DNA conformation, but not relative to the B-DNA conformation. PMID- 11891630 TI - A quantum-dynamics study of the prototropic tautomerism of guanine and its contribution to spontaneous point mutations in Escherichia coli. AB - High-level quantum-chemical and quantum-dynamics calculations are reported on the tautomerization equilibrium and rate constants of guanine and its complexes with one and two water molecules. The results are used to estimate the fraction of guanine present in the cell during DNA synthesis as the unwanted tautomer that forms an irregular base pair with thymine, thus giving rise to a spontaneous GC - > AT point mutation. A comparison of the estimated mutation frequency with the observed frequency in Escherichia coli is used to analyze two proposed mechanisms, differing in the extent of equilibration reached in the tautomerization reaction. In the absence of water, the equilibrium concentration of tautomeric forms is relatively large, but the barrier to their formation is high. If water is present, tautomeric forms are less favored, but water molecules may serve as efficient proton conduits causing rapid tautomerization. It is tentatively concluded that the mechanism in which a high tautomerization barrier keeps the tautomeric transformation far from a state of equilibrium is more likely than a mechanism in which water and/or polymerases produce a low equilibrium concentration of the tautomeric forms. PMID- 11891631 TI - Evaluation of free energy landscape for base-amino acid interactions using ab initio force field and extensive sampling. AB - Structural data of protein-DNA complex show redundancy and flexibility in base amino acid interactions. To understand the origin of the specificity in protein DNA recognition, we calculated the interaction free energy, enthalpy, entropy, and minimum energy maps for AT-Asn, GC-Asn, AT-Ser, and GC-Ser by means of a set of ab initio force field with extensive conformational sampling. We found that the most preferable interactions in these pairs are stabilized by hydrogen bonding, and are mainly enthalpy driven. However, minima in the free energy maps are not necessarily the same as those in the minimum energy map or enthalpy maps, due to the entropic effect. The effect of entropy is particularly important in the case of GC-Asn. Experimentally observed structures of base-amino acid interactions are within preferable regions in the calculated free energy maps, where there are many different interaction configurations with similar energy. The full geometry optimization procedure using ab initio molecular orbital method was applied to get the optimal interaction geometries for AT-Asn, GC-Asn, AT-Ser, and GC-Ser. We found that there are various base-amino acid combinations with similar interaction energies. These results suggest that the redundancy and conformational flexibility in the base-amino acid interactions play an important role in the protein-DNA recognition. PMID- 11891632 TI - Converging methods in developmental science: an introduction. AB - This special issue of Developmental Psychobiology reflects a number of recent advances in the field of developmental neuroscience. The most evident are methodological advances in noninvasive neuroimaging such as those described in a parallel special issue of Developmental Science, Volume 5, 2002. While advances in imaging methods offer a new era in developmental research, other methods (e.g., animal, computational, lesion, and genetic) remain essential in constraining and informing theories of brain and behavioral development. The papers in this issue highlight the importance of a converging methods approach to the study of developmental science and illustrate how a variety of available tools allow insights into both new and classic developmental questions. PMID- 11891633 TI - Developing a brain specialized for face perception: a converging methods approach. AB - Adults are normally very quick and accurate at recognizing facial identity. This skill has been explained by two opposing views as being due to the existence of an innate cortical "face module" or to the natural consequence of adults' extensive experience with faces. Neither of these views puts particular importance on studying development of face-processing skills, as in one view the module simply comes on-line and in the other view development is equated with adult learning. In this article, we present evidence from a variety of methodologies to argue for an "interactive specialization" view. In this view, orienting tendencies present early in life ensure faces are a frequent visual input to higher cortical areas in the ventral visual pathway. In this way, cortical specialization for face processing is an emergent product of the interaction of factors both intrinsic and extrinsic to the developing child. PMID- 11891634 TI - Social interest and the development of cortical face specialization: what autism teaches us about face processing. AB - Investigations of face processing in persons with an autism spectrum disorder (ASD) inform upon theories of the development of "normal" face processing, and the story that emerges challenges some models of the nature and origin of cortical face specialization. Individuals with an ASD possess deficits in face processing and a lack of a fusiform face area (FFA). Evidence from studies of ASD can be conceptualized best using an expertise framework of face processing rather than models that postulate a face module in the fusiform gyrus. Because persons with an ASD have reduced social interest, they may fail to develop cortical face specialization. Face specialization may develop in normal individuals because they are socially motivated to regard the face, and such motivation promotes expertise for faces. The amygdala is likely the key node in the system that marks objects as emotionally salient and could be crucial to the development of cortical face specialization. PMID- 11891635 TI - Testing neural models of the development of infant visual attention. AB - Several models of the development of infant visual attention have used information about neural development. Most of these models have been based on nonhuman animal studies and have relied on indirect measures of neural development in human infants. This article discusses methods for studying a "neurodevelopmental" model of infant visual attention using indirect and direct measures of cortical activity. We concentrate on the effect of attention on eye movement control and show how animal-based models, indirect measurement in human infants, and direct measurement of brain activity inform this model. PMID- 11891636 TI - Clinical, imaging, lesion, and genetic approaches toward a model of cognitive control. AB - The ability to suppress or override competing attentional and behavioral responses is a key component of cognitive processes. This ability continues to develop throughout childhood and appears to be disrupted in a number of childhood disorders (e.g., attention deficit/hyperactivity disorder and Tourette syndrome). At least two brain regions have been implicated repeatedly in these disorders- the frontal lobes and the basal ganglia. The common problem in cognitive control and overlap in implicated brain regions across disorders suggest a single underlying biological mechanism. At the same time, the distinct symptomatology observed across these disorders suggests multiple mechanisms are at play. This article presents converging evidence from clinical, neuroimaging, lesion, and genetic studies to provide a mechanistic model of cognitive control whereby the basal ganglia are involved in inhibition of competing actions and the frontal cortex is involved in representing the relevant thoughts and guiding the appropriate behaviors. PMID- 11891637 TI - Active versus latent representations: a neural network model of perseveration, dissociation, and decalage. AB - Children of different ages often perseverate, repeating previous behaviors when they are no longer appropriate, despite appearing to know what they should be doing. Using neural network models, we explore an account of these phenomena based on a distinction between active memory (subserved by the prefrontal cortex) and latent memory (subserved by posterior cortex). The models demonstrate how (a) perseveration occurs when an active memory of currently relevant knowledge is insufficiently strong to overcome a latent bias established by previous experience, (b) apparent dissociations between children's knowledge and action may reflect differences in the amount of conflict between active and latent memories that children need to resolve in the tasks, and (c) differences in when children master formally similar tasks (decalage) may result from differences in the strength of children's initial biases. The models help to clarify how prefrontal development may lead to advances in flexible thinking. PMID- 11891638 TI - The lesion methodology: contrasting views from adult and child studies. AB - A traditional approach for examining brain-behavior relations has been the lesion method. This method assumes a direct correspondence between the cognitive process compromised and the site of lesion. Historically, studies with adults have used this framework to map brain functions. In contrast, studies of children with early injury have addressed quite different issues. Developmental animal lesion studies and pediatric neuropsychology studies have focused on the level of plasticity exhibited following early injury. Resilency in behavioral development has suggested change in the underlying neural substrate. A new set of studies has applied converging, MRI-based methods to examine anatomical and functional development in intact brain regions following early injury and compared these data with behavioral outcomes on the same children. The findings reveal an interaction between early injury and normal mechanisms of development, which manifest as atypical behavioral, structural, and functional development. PMID- 11891640 TI - Language, gesture, and the developing brain. AB - Do language abilities develop in isolation? Are they mediated by a unique neural substrate, a "mental organ" devoted exclusively to language? Or is language built upon more general abilities, shared with other cognitive domains, and mediated by common neural systems? Here, we review results suggesting that language and gesture are "close family", then turn to evidence that raises questions about how real those "family resemblances" are, summarizing dissociations from our developmental studies of several different child populations. We then examine both these veins of evidence in light of some new findings from the adult neuroimaging literature and suggest a possible reinterpretation of these dissociations as well as new directions for research with both children and adults. PMID- 11891641 TI - Different approaches to relating genotype to phenotype in developmental disorders. AB - In this article, we discuss the complex problem of relating genotype to phenotype and challenge the simple mapping of genes to higher level cognitive modules. We examine various methods that have been used to investigate this relation including quantitative genetics, molecular genetics, animal models, and in-depth psychological and computational studies of developmental disorders. Both single gene and multiple gene disorders indicate that the relationship between genotype and phenotype is very indirect and that, rather than identifying mere snapshots of developmental outcomes, the process of ontogenetic development itself must be taken into account. PMID- 11891642 TI - A converging-methods approach to fragile X syndrome. AB - Converging approaches across domains of brain anatomy, cell biology, and behavior indicate that Fragile X syndrome, arising from impaired expression of a single gene and protein, appears to involve an aberration of normal developmental processes. Synapse overproduction and selective elimination, or pruning, characterize normal brain development. In autopsy tissue from Fragile X patients and in a knockout mouse model of the disease, synapse overproduction appears to occur unaccompanied by synapse pruning and maturation, leaving an excess of immature spine synapses in place. The absence of the Fragile X protein seems to impair the synthesis of important proteins at synapses. The developmental outcome in Fragile X is a nervous system that is relatively disorganized, resulting in disrupted perceptual, and cognitive social, behavior. PMID- 11891643 TI - Convergence of psychological and biological development. AB - The special issue provides an opportunity to assess the degree of integration that has been achieved between studies of psychological and biological development. This commentary uses the material in the special issue to address integration of the two fields and to suggest some future developments. PMID- 11891639 TI - The importance of rapid auditory processing abilities to early language development: evidence from converging methodologies. AB - The ability to process two or more rapidly presented, successive, auditory stimuli is believed to underlie successful language acquisition. Likewise, deficits in rapid auditory processing of both verbal and nonverbal stimuli are characteristic of individuals with developmental language disorders such as Specific Language Impairment. Auditory processing abilities are well developed in infancy, and thus such deficits should be detectable in infants. In the studies presented here, converging methodologies are used to examine such abilities in infants with and without a family history of language disorder. Behavioral measures, including assessments of infant information processing, and an EEG/event-related potential (ERP) paradigm are used concurrently. Results suggest that rapid auditory processing skills differ as a function of family history and are predictive of later language outcome. Further, these paradigms may prove to be sensitive tools for identifying children with poor processing skills in infancy and thus at a higher risk for developing a language disorder. PMID- 11891644 TI - Excitotoxic and metabolic damage to the rodent striatum: role of the P75 neurotrophin receptor and glial progenitors. AB - After injury, the striatum displays several morphologic responses that may play a role in both regenerative and degenerative events. One such response is the de novo expression of the low-affinity p75 neurotrophin receptor (p75(NTR)), a gene that plays critical roles in central nervous system (CNS) cell death pathways. The present series of experiments sought to elucidate the cellular origins of this p75(NTR) response, to define the conditions under which p75(NTR) is expressed after striatal injury, and how this receptor expression is associated with neuronal plasticity. After chemical lesions, by using either the excitotoxin quinolinic acid (QA) or the complex II mitochondria inhibitor 3-nitropropionic acid (3-NP), we compared the expression of the p75(NTR) receptor within the rat striatum at different survival times. Intrastriatal administration of QA between 7 days and 21 days postlesion induced p75(NTR) expression in astrocytes that was preferentially distributed throughout the lesion core. P75(NTR) immunoreactivity within astrocytes was seen at high (100-220 nmol) but not low (50 nmol) QA doses. Seven and 21 days after 3-NP lesions, p75(NTR) expression was present in astrocytes at all doses tested (100-1,000 nmol). However, in contrast to QA, these cells were located primarily around the periphery of the lesion and not within the lesion core. At the light microscopic level p75(NTR) immunoreactive elements resembled vasculature: but did not colocalize with the pan endothelium cell marker RecA-1. In contrast, p75(NTR)-containing astrocytes colocalized with nestin, vimentin, and 5-bromo-2-deoxyuridine, indicating that these cells are newly born astrocytes. Additionally, striatal cholinergic neurons were distributed around the lesion core expressed p75(NTR) 3-5 days after lesion in both QA and 3-NP lesions. These cells did not coexpress the pro-apoptotic degradation enzyme caspase-3. Taken together, these data indicate that striatal lesions created by means of excitotoxic or metabolic mechanisms trigger the expression of p75(NTR) in structures related to progenitor cells. The expression of the p75(NTR) receptor after these chemical lesions support the concept that this receptor plays a role in the initiation of endogenous cellular events associated with CNS injury. PMID- 11891645 TI - Differential expression of nm23 genes in adult mouse dorsal root ganglia. AB - Nm23 has been identified as a gene family encoding different isoforms of nucleoside diphosphate kinase (NDPK). This protein is a key enzyme in nucleotide metabolism and has been shown to play important roles in various cellular functions. In the present study, we have investigated the expression of three isotypes in mouse dorsal root ganglia. In situ hybridization and reverse transcriptase-polymerase chain reaction analysis demonstrated high levels of nm23 M1, -M2, and -M3 mRNA expression in peripheral nervous tissue. Moreover, in situ hybridization also displayed a specific nuclear localization for nm23-M2 mRNA. Immunohistochemistry with light and electron microscopy on isoform-specific antibodies revealed a differential subcellular distribution of NDPK isoforms. Isoform A was mainly cytosolic, showing only partial association with organelles. In contrast, isoform B was also found in the nucleus, which is in agreement with its proposed role as a transcription factor. The results also indicate a preferential association of isoform C with endoplasmic reticulum and plasma membranes in neuronal cells. Furthermore, isoform C appeared to combine with other NDPK isoforms as demonstrated by double-labeling evidence by electron microscopy and might be responsible for binding NDPK oligomers to membranes. Thus, isoform C may be considered as a protein of importance for maintaining intracellular pools of GTP in the vicinity of membranes and, hence, for transmembrane signaling. The results indicate a high expression of NDPK isoforms, not only in the central but also in the peripheral nervous system. Their different subcellular compartmentalization suggests that they have isoform specific roles in neuronal cell physiology. PMID- 11891646 TI - Firing properties of axotomized central nervous system neurons recover after graft reinnervation. AB - Axotomy produces changes in the electrical properties of neurons and in their synaptic inputs, leading to alterations in firing pattern. We have considered the possibility that these changes occur as a result of the target deprivation induced by the lesion. Thus, we have provided a novel target to axotomized central neurons by grafting embryonic tissue at the lesion site to study the target dependence of discharge characteristics. The extracellular single-unit electrical activity of abducens internuclear neurons was recorded in the alert behaving cat in control, after axotomy, and after axotomy plus the implantation of cerebellar primordium. As recently characterized (de la Cruz et al. [2000] J. Comp. Neurol. 427:391-404), firing alterations induced by axotomy included an overall decrease in firing rate and a loss of eye-related signals, i.e., eye position and velocity neuronal sensitivities, that do not resume to normality with time. The grafting of a novel target to the injured abducens internuclear neurons restored the normal firing and sensitivities as recorded in the majority of units. To study the reinnervation of the implant, we performed anterograde labeling with biocytin combined with electron microscopy visualization. Axons of abducens internuclear neurons grew into the transplant sprouting into granule cell and molecular layers, as characterized by the immunostaining for gamma aminobutyric acid and calbindin D-28k. Ultrastructural examination of labeled axons and boutons revealed the establishment of synaptic contacts, mainly axodendritic, with different cell types of the grafted cerebellar cortex. Therefore, these data indicate that axotomized central neurons resume to normal firing after the reinnervation of a novel target. PMID- 11891647 TI - Hooded sensilla homologues: structural variations of a widely distributed bimodal chemomechanosensillum. AB - A diversity of sensilla has been described in crustaceans, both across species and within a given species. However, few homologous setal types have been identified in crustaceans. In this study we examined setae with features of the hooded sensillum, which is a class of bimodal chemomechanosensilla first identified on antennules of the Caribbean spiny lobster Panulirus argus. We examined the antennules of 13 species representing seven families of malacostracan crustaceans, and most body surfaces of P. argus, and compared the sensillar morphology from different species and from different body regions to identify interspecific and intraspecific homologues of hooded sensilla. Our results show that sensilla with morphological characteristics of antennular hooded sensilla are present and have a similar pattern of distribution on the antennules of reptantian species representing three families (Palinuridae and Scyllaridae of the Achelata and Nephropidae of the Homarida). Furthermore, hooded sensillar homologues are present on most body surfaces of P. argus. However, there are intraspecific and interspecific variations in the morphology of these sensilla. We present evidence that supports the idea that postembryonic changes in individual sensilla may be responsible for some of these morphological variations. Despite these variations, we conclude that the sensilla are homologues, because they have several common characteristics, similar positions on the body surface, similar substructures, a continuum to their morphological variations, and morphological variation that is correlated with phylogenetic similarity. Taken together these results support the idea that the hooded sensillum is a singular and biologically important sensillar type that has a broad distribution. PMID- 11891648 TI - Nociceptin/orphanin FQ and opioid receptor-like receptor mRNA expression in dopamine systems. AB - Although nociceptin/orphanin FQ (N/OFQ) influences dopamine (DA) neuronal activity, it is not known whether N/OFQ acts directly on DA neurons, indirectly by means of local circuitry, or both. We used two parallel approaches, dual in situ hybridization (ISH) and neurotoxic lesions of DA neurons by using 6 hydroxydopamine (6-OHDA), to ascertain whether N/OFQ and the N/OFQ receptor (NOP) mRNA are expressed in DA neurons in the ventral tegmental area (VTA) and substantia nigra compacta (SNc). In the VTA and SNc, small populations (approximately 6-10%) of N/OFQ-containing neurons coexpressed mRNA for tyrosine hydroxylase (TH), the rate-limiting enzyme for DA synthesis. Similarly, very few (1-2%) TH-positive neurons contained N/OFQ mRNA signal. A majority of NOP positive neurons (approximately 75%) expressed TH mRNA and roughly half of the TH containing neurons expressed NOP mRNA. Many N/OFQ neurons (approximately 50-60%) expressed glutamic acid decarboxylase 65 and 67 mRNAs, markers for gamma aminobutyric acid (GABA) neurons. In the 6-OHDA lesion studies, NOP mRNA levels were nearly 80 and 85% lower in the VTA and SNc, respectively, on the lesioned side. These lesions appear to lead to compensatory changes, with N/OFQ mRNA levels approximately 60% and 300% higher in the VTA and SNc, respectively, after 6-OHDA lesions. Finally, N/OFQ-stimulated [(35)S]guanylyl-5'-O-(gamma-thio) triphosphate levels were decreased in the VTA and SNc but not the prefrontal cortex after 6-OHDA lesions. Accordingly, it appears that N/OFQ mRNA was found largely on nondopaminergic (i.e., GABA) neurons, whereas NOP mRNA was located on DA neurons. N/OFQ is in a position to influence DA neuronal activity by means of the NOP located on DA neurons. PMID- 11891649 TI - Differential distribution of spermidine/spermine-like immunoreactivity in neurons of the adult rat brain. AB - The polyamines spermidine and spermine are small, widely distributed polycations. In the brain, they confer rectification properties upon inwardly rectifying potassium channels and Ca(2+)-permeable alpha-amino-3-hydroxy-5-methyl-4 isoxazole-propionate (AMPA)/kainate receptors and also modify functional properties of N-methyl-D-aspartate receptors. Therefore, functional roles of spermidine/spermine in the adult brain will depend on the colocalization of the spermidine/spermine-sensitive receptors/channels and the polyamines either in the same or in closely associated cell types. We previously immunocytochemically demonstrated a prominent localization of spermidine/spermine in glial cells, especially astrocytes (Laube and Veh [ 1997] Glia 19:171-179). In contrast to the commonly accepted assumption of a ubiquitous distribution of polyamines in various cell types, in neurons of the rat brain, we detected a highly diverse spermidine/spermine-like immunoreactivity. The immunoreactivity in neurons and neuropil throughout the rat brain is listed according to intensity in arbitrary groups. The strongest neuronal staining was observed in the hypothalamic paraventricular, supraoptic, and accessory neurosecretory nuclei. Strong cytoplasmic staining was also evident in some motor and somatosensory areas such as the Me5 nucleus of the mesencephalic trigeminal tract, the nucleus ruber, and the large motor neurons of the spinal cord ventral horn. In contrast, in most cortical and hippocampal regions spermidine/spermine-like immunoreactivity in neurons was relatively weak, whereas in these areas, the labeling pattern was dominated by a diffuse neuropil labeling. In addition to spermidine/spermine immunocytochemistry, ornithine decarboxylase labeling was performed and the resulting labeling patterns were compared. The prominent localization of spermidine/spermine in neurosecretory neurons might point to a functional role different from channel/receptor modification. In these neurons, polyamines might be involved in secretory processes. PMID- 11891650 TI - Distribution of two splice variants of the glutamate transporter GLT1 in the retinas of humans, monkeys, rabbits, rats, cats, and chickens. AB - Antibodies have been generated against two carboxyl-terminal splice variants of the glutamate transporter GLT1, namely, the originally described version of GLT1 and GLT1-B, and labelling has been examined in multiple species, including chickens and humans. Although strong specific labelling was observed in each species, divergent patterns of expression were noted. Moreover, each antibody was sensitive to the phosphorylation state of the appropriate protein, because chemical removal of phosphates using alkaline phosphatase revealed a broader range of labelled elements in most cases. In general, GLT1-B was present in cone photoreceptors and in rod and cone bipolar cells in the retinas of rabbits, rats, and cats. In the cone-dominated retinas of chickens and in marmosets, GLT1-B was associated only with cone photoreceptors, whereas, in macaque and human retinas, GLT1-B was associated with bipolar cells and terminals of photoreceptors. In some species, such as cats, GLT-B was also present in horizontal cells. By contrast, GLT1 distribution varied. GLT1 was associated with amacrine cells in chickens, rats, cats, and rabbits and with bipolar cells in marmosets and macaques. In the rat retina, rod photoreceptor terminals also contained GLT1, but this was evident only in enzymatically dephosphorylated tissues. We conclude that the two variants of GLT1 are present in all species examined but are differentially distributed in a species-specific manner. Moreover, each cell type generally expresses only one splice variant of GLT1. PMID- 11891651 TI - Effects of age on the glial cells in the rhesus monkey optic nerve. AB - The optic nerve is a circumscribed white matter tract consisting of myelinated nerve fibers and neuroglial cells. Previous work has shown that during normal aging in the rhesus monkey, many optic nerves lose some of their nerve fibers, and in all old optic nerves there are both myelin abnormalities and degenerating nerve fibers. The present study assesses how the neuroglial cell population of the optic nerve is affected by age. To address this question, optic nerves from young (4-10 years) and old (27-33 years) rhesus monkeys were examined by using both light and electron microscopy. It was found that with age the astrocytes, oligodendrocytes, and microglia all develop characteristic cytoplasmic inclusions. The astrocytes hypertrophy and fill space vacated by degenerated nerve fibers, and they often develop abundant glial filaments in their processes. Oligodendrocytes and microglial cells both become more numerous with age, and microglial cells often become engorged with phagocytosed debris. Some of the debris can be recognized as degenerating myelin, and in general, the greater the loss of nerve fibers, the more active the microglial cells become. PMID- 11891652 TI - Colocalization of gamma-aminobutyric acid-like immunoreactivity and catecholamines in the feeding network of Aplysia californica. AB - Functional consequences of neurotransmitter coexistence and cotransmission can be readily studied in certain experimentally favorable invertebrate motor systems. In this study, whole-mount histochemical methods were used to identify neurons in which gamma-aminobutyric acid (GABA)-like immunoreactivity (GABAli) was colocalized with catecholamine histofluorescence (CAh; FaGlu method) and tyrosine hydroxylase (TH)-like immunoreactivity (THli) in the feeding motor circuitry (buccal and cerebral ganglia) of the marine mollusc Aplysia californica. In agreement with previous reports, five neurons in the buccal ganglia were found to exhibit CAh. These included the paired B20 buccal-cerebral interneurons (BCIs), the paired B65 buccal interneurons, and an unpaired cell with projections to both cerebral-buccal connectives (CBCs). Experiments in which the FaGlu method was combined with the immunohistochemical detection of GABA revealed double labeling of all five of these neurons. An antibody generated against TH, the rate-limiting enzyme in the biosynthesis of catecholamines, was used to obtain an independent determination of GABA-CA colocalization. Biocytin backfills of the CBC performed in conjunction with TH immunohistochemistry revealed labeling of the rostral B20 cell pair and the unpaired CBI near the caudal surface of the right hemiganglion. THli was also present in a prominent bilateral pair of caudal neurons that were not stained with CBC backfills. On the basis of their position, size, shape, and lack of CBC projections, the lateral THli neurons were identified as B65. Double labeling immunohistochemical experiments revealed GABAli in all five buccal THli neurons. Finally, GABAli was observed in individual B20 and B65 neurons that were identified using electrophysiological criteria and injected with a marker (neurobiotin). Similar methods were used to demonstrate that a previously identified catecholaminergic cerebral-buccal interneuron (CBI) designated CBI-1 contained THli but did not contain GABAli. Although numerous THli and GABAli neurons and fibers were present in the cerebral and buccal ganglia, additional instances of their colocalization were not observed. These findings indicate that GABA and a catecholamine (probably dopamine) are colocalized in a limited number of interneurons within the central pattern generator circuits that control feeding-related behaviors in Aplysia. PMID- 11891653 TI - Nucleus retroambiguus projections to the periaqueductal gray in the cat. AB - The nucleus retroambiguus (NRA) of the caudal medulla is a relay nucleus by which neurons of the mesencephalic periaqueductal gray (PAG) reach motoneurons of pharynx, larynx, soft palate, intercostal and abdominal muscles, and several muscles of the hindlimbs. These PAG-NRA-motoneuronal projections are thought to play a role in survival behaviors, such as vocalization and mating behavior. In the present combined antero- and retrograde tracing study in the cat, we sought to determine whether the NRA, apart from the neurons projecting to motoneurons, also contains cells projecting back to the PAG. After injections of WGA-HRP in the caudal and intermediate PAG, labeled neurons were observed in the NRA, with a slight contralateral preponderance. In contrast, after injections in the rostral PAG or adjacent deep tectal layers, no or very few labeled neurons were present in the NRA. After injection of [(3)H]leucine in the NRA, anterograde labeling was present in the most caudal ventrolateral and dorsolateral PAG, and slightly more rostrally in the lateral PAG, mainly contralaterally. When the [(3)H]leucine injection site extended medially into the medullary lateral tegmental field, labeling was found in most parts of the PAG as well as in the adjoining deep tectal layers. No labeled fibers were found in the dorsolateral PAG, and only a few were found in the rostral PAG. Because the termination pattern of the NRA fibers in the PAG overlaps with that of the sacral cord projections to the PAG, it is suggested that the NRA-PAG projections play a role in the control of motor functions related to mating behavior. PMID- 11891654 TI - Ontogeny of NADPH diaphorase/nitric oxide synthase reactivity in the brain of Xenopus laevis. AB - The development of nitric oxide synthase (NOS) expression in the brain of Xenopus laevis tadpoles was studied by means of immunohistochemistry using specific antibodies against NOS and enzyme histochemistry for nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase. Both techniques yielded identical results and were equally suitable for demonstrating the nitrergic system in the brain. The only mismatches were observed in the olfactory nerve and glomeruli and in the terminal nerve; they were intensely labeled with the NADPH-diaphorase technique but failed to stain with NOS immunohistochemistry. As early as stage 33, nitrergic cells were observed in the caudal rhombencephalon within the developing inferior reticular nucleus. At later embryonic stages, different sets of reticular and tegmental neurons were labeled in the middle reticular nucleus and, more conspicuously, in the laterodorsal and pedunculopontine tegmental nuclei. As development proceeded, new nitrergic cell groups gradually appeared in the mesencephalon, diencephalon, and telencephalon. A general caudorostral temporal sequence was observed, both in the whole brain and within each main brain subdivision. The premetamorphic period was mainly characterized by the maturation of the cell populations developed in the embryonic period. During prometamorphosis, the nitrergic system reached an enormous development, and many new cell groups were observed for the first time, in particular in the telencephalon. By the climax of metamorphosis, the pattern of organization of nitrergic cells and fibers observed in the brain was similar to that present in the adult brain. Transient expression of NOS was not detected in any brain region. Our data suggest that nitric oxide plays an important role during brain development of Xenopus. Comparison with the developmental pattern of nitrergic systems in other vertebrates shows that amphibians possess more common features with amniotes than with anamniotes. PMID- 11891655 TI - Cell types and response properties of neurons in the ventral division of the medial geniculate body of the rabbit. AB - Although there is evidence for multiple classes of thalamic relay neurons in the auditory thalamus, correlative anatomical and physiological studies are lacking. We have used the juxtacellular labeling technique, in conjunction with Nissl, Golgi, and immunocytochemical methods, to study the morphology and response properties of cells in the ventral division of the medial geniculate body of the rabbit. Single units in the ventral division of the medial geniculate body (MGV) were characterized extracellularly with monaural and binaural tone and noise bursts (100- to 250-msec duration). Characterized units were filled with biocytin and visualized with an antibody enhanced diaminobenzidine reaction. A total of 31 neurons were physiologically characterized and labeled with the juxtacellular technique. Labeled neurons were fully reconstructed from serial sections by using a computer microscope system. Three subregions of the rabbit MGV were identified, each characterized by differences in Nissl architecture, calcium-binding protein expression, and by the dendritic orientation of tufted relay neurons. In general, the dendritic fields of relay neurons were closely aligned with the cellular laminae. Qualitative and quantitative analyses revealed two types of presumptive relay neurons within the MGV. Type I cells had thick dendrites with a greater total volume and morphologically diverse appendages compared with the Type II cells whose dendrites were thin with a moderate number of small spines. Both classes were acoustically responsive and exhibited a variety of response patterns, including onset, offset, and sustained responses. In terms of binaural characteristics, most (ca. 53%) labeled neurons were of the EE type, with the remaining cells classified as EO (27%) or EI (20%) response types. Two types of presumptive interneurons were also seen: bipolar neurons with large dendritic fields and a small neurogliaform variety. Cell types and dendritic orientation within the MGV are discussed in terms of the physiological organization of the rabbit auditory thalamus. PMID- 11891656 TI - Morphology and projection pattern of medial and dorsal pallial neurons in the frog Discoglossus pictus and the salamander Plethodon jordani. AB - In the frog Discoglossus pictus and the salamander Plethodon jordani, the morphology and axonal projection pattern of neurons in the medial and dorsal pallium were determined by intracellular biocytin labeling. A total of 77 pallial neurons were labeled in the frog and 58 pallial neurons in the salamander. Within the medial pallium (MP) of the frog, four types of neurons were identified on the basis of differences in their axonal projection pattern. Type I neurons have bilateral projections into telencephalic and diencephalic areas; type II neurons have bilateral projections to telencephalic areas and ipsilaterally descending projections to diencephalic regions; type III neurons have only intratelencephalic connections, and a single type IV neuron has ipsilaterally descending projections. The somata of the four types occupy four nonoverlapping zones. Neurons of the dorsal pallium (DP) project exclusively to the ipsilateral MP and to the dorsal edge of the lateral pallium. In the ventral MP of the salamander, neurons have mostly intratelencephalic projections. Neurons in the dorsal MP project bilaterally to diencephalic and telencephalic regions. Neurons in the medial DP project ipsilaterally to the MP, lateral septum, nucleus accumbens, medial amygdala, and the internal granule layer of the olfactory bulb. In five cases, fibers were found in the commissura hippocampi, but in only two cases could these fibers be followed toward the contralateral MP and septum. Neurons in the lateral DP had no contralateral projections; they projected to the ipsilateral MP and in eight cases to the ipsilateral septum as well. Based on similarities of cytoarchitecture and projection pattern in neurons of the MP and DP, it is proposed that both frogs and salamanders have an MP subdivided into a ventral and dorsal portion, and a DP subdivided into a medial and a lateral portion. PMID- 11891657 TI - Differential expression of alpha 3 and alpha 6 integrins in the developing mouse inner ear. AB - The development of the mammalian inner ear involves a complex series of cell-cell and cell-extracellular matrix interactions. These interactions are likely to be mediated by families of adhesion molecules, including the integrins. We have studied the expression of three integrin subunits known to be expressed on epithelia in a number of tissues (namely, alpha3, alpha6, and beta4) during the development of the murine inner ear. At E10.5, both alpha3 and alpha6 were expressed in the epithelial layers of the otocyst. The expression of alpha6 was concentrated in an anterioventral region of the epithelium and in a proportion of the cells forming the cochlear-vestibular and facial ganglia. By E12.5, alpha6 showed a more restricted expression, confined mainly to the pro-sensory epithelia and the neural processes from the cochlear-vestibular ganglion. In contrast, alpha3 was expressed in epithelia adjacent to the pro-sensory areas. This reciprocal expression pattern was maintained until birth. Between birth and P6, a switch in expression occurred such that alpha3 was upregulated and alpha6 was downregulated in the sensory epithelia of both the auditory and vestibular systems. At this stage, alpha3 was expressed in all the epithelia lining the scala media, thus defining the endolymph compartment. The expression of beta4 was restricted to epithelial/mesenchymal borders throughout the developmental stages studied, suggesting that alpha6 expression observed within the epithelium and neuronal tissue was alpha6beta1. The early expression and changing pattern of alpha3 and alpha6 integrins during development of the mammalian inner ear suggests that they may be involved in the molecular processes that define epithelial boundaries and guide sensory innervation. PMID- 11891658 TI - Organization of cutaneous ventrodorsal and rostrocaudal axial lines in the rat hindlimb and trunk in the dorsal horn of the spinal cord. AB - The somatotopic organization of cutaneous primary afferents projecting to the dorsal horn of the rat spinal cord was investigated. The fluorescent neurotracer, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) was applied to cutaneous incisions made along ventrodorsal axial lines (VDALs) or rostrocaudal axial lines (RCALs) of the trunk and hindlimb. DiI-induced fluorescent zones appeared in laminae I-III of the dorsal horn in the transverse section. Several fluorescent zones appeared at different mediolateral portions after tracer application to VDALs. After tracer was applied to RCALs, a single zone of fluorescence was observed. Serial transverse sections were used to reconstruct fluorescent zones in lamina II and to illustrate the rostrocaudally elongated band-like projection fields in a horizontal plane. In the horizontal plane, the fluorescent zones of VDALs were reconstructed to band-like projection fields. These fields were arranged mediolaterally and extended rostrocaudally for approximately the length of one spinal cord segment or less. The fluorescent zones of RCALs were reconstructed to one band-like projection field. This field extended rostrocaudally over several spinal cord segments. Cutaneous afferents from the ventral median line of the trunk, tail, hindlimb, sole, and ventral side of the digits projected to the medial margin of the dorsal horn. Cutaneous afferents from the dorsal median lines projected to the lateral margin of the dorsal horn. By analyzing the pattern of the body surface regions and the VDALs and RCALs, the central projection fields of body surface regions could be hypothesized, based on the central projection fields of the individual VDAL and RCAL afferents. Thus, we established a detailed dorsal view map of the central projection fields of cutaneous primary afferents. PMID- 11891659 TI - Noradrenergic innervation of the developing and mature visual and motor cortex of the rat brain: a light and electron microscopic immunocytochemical analysis. AB - The noradrenergic (NA) innervation of the developing and adult visual and motor cortex of the rat was examined with light and electron microscopic immunocytochemistry by using antibodies against dopamine-beta-hydroxylase. At birth, NA fibers were present in both cortical areas, appearing as two tangential streams, one above and the other below the cortical plate. During the subsequent weeks, these two streams arborized gradually innervating all cortical layers. The adult pattern of distribution was attained by postnatal day 14, but the density of innervation, which was higher in the motor than in the visual cortex, appeared similar to the adult by the end of the third postnatal week. Electron microscopic analysis revealed that a low proportion of NA varicosities (the highest value was 12% in the adult motor cortex in single sections) were engaged in synaptic contact, throughout development, in both areas examined. The overwhelming majority of these synapses were symmetrical, involving predominantly small or medium dendrites. This evidence suggests that transmission by diffusion is the major mode of NA action in the developing and adult cerebral cortex. Noradrenaline released in the rare synaptic junctions may act mainly to reduce the activity of its cortical targets. The results altogether provide morphologic evidence for an involvement of noradrenaline in the development of the neocortex and, along with earlier data on the serotonergic system, indicate that the monoaminergic systems are endowed with a specific anatomic organization in various areas of the brain. PMID- 11891660 TI - Cholinergic innervation of the chick basilar papilla. AB - Antibodies directed against choline acetyltransferase (ChAT), the synthesizing enzyme for acetylcholine (ACh) and a specific marker of cholinergic neurons, were used to label axons and nerve terminals of efferent fibers that innervate the chick basilar papilla (BP). Two morphologically distinct populations of cholinergic fibers were labeled and classified according to the region of the BP they innervated. The inferior efferent system was composed of thick fibers that coursed radially across the basilar membrane in small fascicles, gave off small branches that innervated short hair cells with large cup-like endings, and continued past the inferior edge of the BP to ramify extensively in the hyaline cell area. The superior efferent system was made up of a group of thin fibers that remained in the superior half of the epithelium and innervated tall hair cells with bouton endings. Both inferior and superior efferent fibers richly innervated the basal two thirds of the BP. However, the apical quarter of the chick BP was virtually devoid of efferent innervation except for a few fibers that gave off bouton endings around the peripheral edges. The distribution of ChAT-positive efferent endings appeared very similar to the population of efferent endings that labeled with synapsin antisera. Double labeling with ChAT and synapsin antibodies showed that the two markers colocalized in all nerve terminals that were identified in BP whole-mounts and frozen sections. These results strongly suggest that all of the efferent fibers that innervate the chick BP are cholinergic. PMID- 11891661 TI - Hair cell development in vivo and in vitro: analysis by using a monoclonal antibody specific to hair cells in the chick inner ear. AB - The purpose of this study was to establish a hair cell-specific marker and a convenient explant culture system for developing chick otocysts to facilitate in vivo and in vitro studies focusing on hair cell genesis in the inner ear. To achieve this, a hair cell-specific monoclonal antibody, 2A7, was generated by immunizing chick inner ear tissues to a mouse. Through the use of immunofluorescence and immunoelectron microscopy, it was shown that 2A7 immunoreactivity (2A7-IR) was primarily restricted to the apical region of inner ear hair cells, including stereocilia, kinocilia, apical membrane amongst the extending cilia, and superficial layer of the cuticular plate. Although the 2A7 antibody immunolabeled basically all of the hair cells in the posthatch chick inner ear, two different patterns of 2A7-IR were observed; hair cells located in the striolar region of the utricular macula, which consist of two distinct cell types identifiable on the basis of the type of nerve ending, Type I and II hair cells, showed labeling restricted to the basal end of the hair bundles. On the other hand, hair cells in the extrastriolar region, which are exclusively of Type II, showed labeling extending over virtually the entire length of the bundles. These findings raised the possibility that chick vestibular Type II hair cells, characterized by their bouton-type afferent nerve endings, can be divided into two subpopulations. Analysis of developing inner ear by using the 2A7 antibody revealed that this antibody also recognizes newly differentiated immature hair cells. Thus, the 2A7 antibody is able to recognize both immature and mature hair cells in vivo. The developmental potential of embryonic otocysts in vitro was then assessed by using explant cultures as a model. In this study, conventional otocyst explant cultures were modified by placing the tissues on floating polycarbonate filters on culture media, thereby allowing the easy manipulation of explants. In these cultures, 2A7-positive hair cells were differentiated from dividing precursor cells in vitro on the same schedule as in vivo. Furthermore, it was found that hair cells with both types of 2A7-IR were generated in culture as in vivo, indicating that a maturational process of hair cells also occurred. All these results as presented here suggest that the 2A7 monoclonal antibody as a hair cell-specific marker together with the culture system could be a potential tool in analysis of mechanisms underlying hair cell development. PMID- 11891662 TI - Local calcium changes regulate the length of growth cone filopodia. AB - Previous studies have demonstrated that the free intracellular calcium concentration ([Ca(2+)](i)) in growth cones can act as an important regulator of growth cone behavior. Here we investigated whether there is a spatial and temporal correlation between [Ca(2+)](i) and one particular aspect of growth cone behavior, namely the regulation of growth cone filopodia. Calcium was released from the caged compound NP-EGTA (o-nitrophenyl EGTA tetrapotassium salt) to simulate a signaling event in the form of a transient increase in [Ca(2+)](i). In three different experimental paradigms, we released calcium either globally (within an entire growth cone), regionally (within a small area of the lamellipodium), or locally (within a single filopodium). We demonstrate that global photolysis of NP-EGTA in growth cones caused a transient increase in [Ca(2+)](i) throughout the growth cone and elicited subsequent filopodial elongation that was restricted to the stimulated growth cone. Pharmacological blockage of either calmodulin or the Ca(2+)-dependent phosphatase, calcineurin, inhibited the effect of uncaging calcium, suggesting that these enzymes are acting downstream of calcium. Regional uncaging of calcium in the lamellipodium caused a regional increase in [Ca(2+)](i), but induced filopodial elongation on the entire growth cone. Elevation of [Ca(2+)](i) locally within an individual filopodium resulted in the elongation of only the stimulated filopodium. These findings suggest that the effect of an elevation of [Ca(2+)](i) on filopodial behavior depends on the spatial distribution of the calcium signal. In particular, calcium signals within filopodia can cause filopodial length changes that are likely a first step towards directed filopodial steering events seen during pathfinding in vivo. PMID- 11891664 TI - BMP-2 and cAMP elevation confer locus coeruleus neurons responsiveness to multiple neurotrophic factors. AB - The locus coeruleus (LC) is a major target of several neurodegenerative disorders, including Parkinson's and Alzheimer's diseases. However, very little is known of the trophic requirements of LC neurons. In the present work, we have studied the biological activity of neurotrophic factors from different families in E15 primary cultures of LC neurons. In agreement with previous results, neurotrophin-3 (NT-3) and also glial cell line- derived neurotrophic factor (GDNF) increased the number of embryonic LC noradrenergic neurons in the presence of serum. In serum-free conditions, none of the factors tested, including NT-3, GDNF, neurturin, basic fibroblast growth factor (bFGF), or bone morphogenetic protein-2 (BMP-2), promoted the survival of tyrosine hydroxylase (TH) immunoreactive neurons at 6 days in vitro. However, when BMP-2 was coadministered with any of these factors the number of LC TH-positive neurons increased twofold. Similar results were obtained by cotreatment of LC neurons with forskolin and NT 3, bFGF, or BMP-2. The strongest effect (a fourfold increase in the number of TH positive cells) was induced by cotreatment with forskolin, BMP-2, and GDNF. Thus, our results show that LC neurons require multiple factors for their survival and development, and suggest that activation of LC neurons by bone morphogenetic proteins and cAMP plays a decisive role in conferring noradrenergic neuron responsiveness to several trophic factors. PMID- 11891665 TI - Neural progenitor cells of the neonatal rat anterior subventricular zone express functional GABA(A) receptors. AB - The interneurons of the olfactory bulb arise from precursor cells in the anterior part of the neonatal subventricular zone, the SVZa, and are distinctive in that they possess a neuronal phenotype and yet undergo cell division. To characterize the differentiation of neonatal SVZa progenitor cells, we analyzed the complement of ionotropic neurotransmitter receptors that they express in vitro. For this analysis, we tested the sensitivity of SVZa progenitor cells to gamma-amino-n butyric acid (GABA), adenosine triphosphate (ATP), kainate, N-methyl-D-aspartate (NMDA), and acetylcholine (ACh) after 1 day in vitro. SVZa progenitor cells had chloride currents activated by GABA and muscimol, the GABA(A) receptor-specific agonist, but were insensitive to ATP, kainate, NMDA, and ACh. In addition, GABA- or muscimol-activated chloride currents were blocked nearly completely by 30 microM bicuculline, the GABA(A) receptor-specific antagonist, suggesting that GABA(B) and GABA(C) receptors are absent. Measurements of the chloride reversal potential by gramicidin-perforated patch clamp revealed that currents generated by activation of GABA(A) receptors were inward, and thus, depolarizing. A set of complementary experiments was undertaken to determine by reverse transcription and polymerase chain reaction (RT-PCR) whether SVZa progenitor cells express the messenger RNA (mRNA) coding for glutamic acid decarboxylase 67 (GAD67), used in the synthesis of GABA and for GABA(A) receptor subunits. Both postnatal day (P0) SVZa and olfactory bulb possessed detectable mRNA coding for GAD67. In P0 SVZa, the GABA(A) receptor subunits detected with RT-PCR included alpha 2-4, beta 1-3, and gamma 2S (short form). By comparison, the P0 olfactory bulb expressed all of the subunits detectable in the SVZa and additional subunit mRNAs: alpha 1, alpha 5, gamma 1, gamma 2L (long form), gamma 3, and delta subunit mRNAs. Antibodies recognizing GABA, GAD, and various GABA(A) receptor subunits were used to label SVZa cells harvested from P0-1 rats and cultured for 1 day. The cells were immunoreactive for GABA, GAD, and the GABA(A) receptor subunits alpha 2-5, beta 1 3, and gamma 2. To relate the characteristics of GABA(A) receptors in cultured SVZa precursor cells to particular combinations of subunits, the open reading frames of the dominant subunits detected by RT-PCR (alpha 2-4, beta 3, and gamma 2S) were cloned into a mammalian cell expression vector and different combinations were transfected into Chinese hamster ovary-K1 (CHO-K1) cells. A comparison of the sensitivity to inhibition by zinc of GABA(A) receptors in SVZa precursor cells and in CHO-K1 cells expressing various combinations of recombinant GABA(A) receptor subunits suggested that the gamma 2S subunit was present and functional in the GABA(A) receptor chloride channel complex. Thus, SVZa precursor cells are GABAergic and a subset of the GABA(A) receptor subunits detected in the olfactory bulb was found in the SVZa, as might be expected because SVZa progenitor cells migrate to the bulb as they differentiate. PMID- 11891663 TI - Chronic alterations in serotonin function: dynamic neurochemical properties in agonistic behavior of the crayfish, Orconectes rusticus. AB - The biogenic amine serotonin [5-hydroxytryptamine (5-HT)] has received considerable attention for its role in behavioral phenomena throughout a broad range of invertebrate and vertebrate taxa. Acute 5-HT infusion decreases the likelihood of crayfish to retreat from dominant opponents. The present study reports the biochemical and behavioral effects resulting from chronic treatment with 5-HT-modifying compounds delivered for up to 5 weeks via silastic tube implants. High performance liquid chromatography with electrochemical detection (HPLC-ED) confirmed that 5,7-dihydroxytryptamine (5,7-DHT) effectively reduced 5 HT in all central nervous system (CNS) areas, except brain, while a concurrent accumulation of the compound was observed in all tissues analyzed. Unexpectedly, two different rates of chronic 5-HT treatment did not increase levels of the amine in the CNS. Behaviorally, 5,7-DHT treated crayfish exhibited no significant differences in measures of aggression. Although treatment with 5-HT did not elevate 5-HT content in the CNS, infusion at a slow rate caused animals to escalate more quickly while 5-HT treatment at a faster rate resulted in slower escalation. 5,7-DHT is commonly used in behavioral pharmacology and the present findings suggest its biochemical properties should be more thoroughly examined. Moreover, the apparent presence of powerful compensatory mechanisms indicates our need to adopt an increasingly dynamic view of the serotonergic bases of behavior like crayfish aggression. PMID- 11891666 TI - Striatopallidal neurons are selectively protected by neurturin in an excitotoxic model of Huntington's disease. AB - Excitotoxicity has been involved in the pathogenesis of several neurodegenerative disorders. Using intrastriatal quinolinic acid (QUIN) injection as an animal model of Huntington's disease, we attempt to identify the neurotransmitter phenotype of striatal projection neurons protected by neurturin (NRTN). Control or NRTN-secreting cell lines were grafted in the striatum before QUIN injection and striatal projection neurons were examined by retrograde Fluorogold labeling and in situ hybridization. Intrastriatal grafting of NRTN-secreting cell line selectively prevented the loss of striatopallidal neurons and also the decrease in the mRNA levels for their markers (glutamic acid decarboxylase 67 and preproenkephalin) induced by QUIN, without affecting striatonigral neurons. Thus, our findings show that NRTN is a selective neuroprotective factor for striatopallidal neurons, suggesting that it might be a candidate for the treatment of movement disorders in which this neuronal population is affected. PMID- 11891668 TI - Development and validation of a non-linear IVIVC model for a diltiazem extended release formulation. AB - To develop and validate internally an in vitro-in vivo correlation (IVIVC) for a diltiazem multi-particulate bead extended release formulation. In vitro dissolution of diltiazem capsules was examined using the following methods: USP Apparatus II (paddle) at 100 rpm and USP Apparatus III at 30 dpm. Seven healthy subjects received three diltiazem formulations (90 mg): slow (S), moderate (M), fast (F) releasing and an oral solution (90 mg). Serial blood samples were collected over 48 h and analyzed by a validated HPLC assay using ultraviolet detection. The f(2) metric (similarity factor) was used to analyze the dissolution data. Linear and non-linear (quadratic, cubic, and sigmoid functions) correlation models were developed using pooled fraction dissolved (FRD) and fraction absorbed (FRA) data from various combinations of the formulations. Predicted diltiazem concentrations were obtained by convolution of the in vivo dissolution rates. Prediction errors were estimated for C(max) and AUC to determine the validity of the correlation. Apparatus II using purified water was found to be the most discriminating dissolution method. Significant intersubject (CV%>50) was observed for C(max) and AUC. The quadratic M/F IVIVC model provided a significant relationship between FRD and FRA when using either two or three of the formulations. An average percent prediction error for C(max) and AUC for all formulations was 12.4% and 9.2%, respectively. The prediction errors observed for C(max) and AUC suggest that the predictability of the quadratic IVIVC model is inconclusive, as such, external validation studies are required. PMID- 11891667 TI - Novel UNC-44 AO13 ankyrin is required for axonal guidance in C. elegans, contains six highly repetitive STEP blocks separated by seven potential transmembrane domains, and is localized to neuronal processes and the periphery of neural cell bodies. AB - Conventional ankyrins are cortical cytoskeletal proteins that form an ankyrin spectrin meshwork underlying the plasma membrane. We report here the unusual structure of a novel ankyrin (AO13 ankyrin, 775,369 Da, 6994 aa, pI = 4.45) that is required for proper axonal guidance in Caenorhabditis elegans. AO13 ankyrin contains the ANK repeat and spectrin-binding domains found in other ankyrins, but differs from all others in that the acidic carboxyl region contains six blocks of serine/threonine/glutamic acid/proline rich (STEP) repeats separated by seven hydrophobic domains. The STEP repeat blocks are composed primarily of sequences related to ETTTTTTVTREHFEPED(E/D)X(n)VVESEEYSASGSPVPSE (E/K)DVE(H/R)VI, and the hydrophobic domains contain sequences related to PESGEESDGEGFGSKVLGFAKK[AGMVAGGVVAAPVALAAVGA]KAAYDALKKDDDEE, which includes a potential transmembrane domain (in brackets). Recombinant protein fragments of AO13 ankyrin were used to prepare polyclonal antisera against the spectrin binding domain (AO271 Ab), the conventional ankyrin regulatory domain (AO280 Ab), the AO13 ankyrin STEP domain (AO346 Ab), the AO13 ankyrin STEP + hydrophobic domain (AO289 Ab), and against two carboxyl terminal domain fragments (AO263 Ab and AO327 Ab). Western blot analysis with these Ab probes demonstrated multiple protein isoforms. By immunofluorescence microscopy, the antispectrin-binding and regulatory domain (AO271 and AO280) antibodies recognized many cell types, including neurons, and stained the junctions between cells. The AO13 ankyrin specific (AO289 and AO346) antibodies showed a neurally restricted pattern, staining nerve processes and the periphery of neural cell bodies. These results are consistent with a role for AO13 ankyrin in neural development. PMID- 11891669 TI - Stereoselective halofantrine and desbutylhalofantrine disposition in the rat: cardiac and plasma concentrations and plasma protein binding. AB - Halofantrine (HF) is a chiral antimalarial drug known to cause cardiac arrhythmias in susceptible patients. In this study, the cardiac uptake and plasma protein binding of HF and desbutylhalofantrine (DHF) enantiomers were examined in the rat. Rats were given 2 mg/kg of either HF HCl or DHF HCl intravenously, then sacrificed at various times after dosing. Specimens were assayed using stereospecific methods. Uptake of HF and DHF enantiomers into heart was rapid. Substantial concentrations of both HF and DHF enantiomers were observed in rat heart, with stereoselectivity being noted for both in plasma and heart. Stereoselectivity was more pronounced for HF (AUC (+):(-) ratio= 1.58) than DHF (AUC (+):(-) ratio =1.16) in heart tissue. Heart:plasma AUC ratios of 6.8-8.0, and 9.3-21, were observed for HF and DHF enantiomers, respectively, indicating that DHF has greater cardiac uptake than HF itself. Plasma protein binding was extensive for both HF and DHF (>99.95%), and was stereoselective for DHF, with a 38% higher unbound fraction for (-)-DHF than antipode. In contrast, binding of HF enantiomers was non-stereoselective. The lower degree of stereoselectivity for DHF in heart tissues was attributable to its greater stereoselectivity in plasma protein binding. PMID- 11891670 TI - Pharmacokinetics of controlled-release verapamil in healthy volunteers and patients with hypertension or angina. AB - AIMS: To study the dose-ranging population pharmacokinetics of controlled release verapamil in healthy subjects and patients with angina or hypertension. To characterize the pharmacodynamics of controlled-release verapamil in patients with hypertension. METHODS: Dose-ranging studies were conducted in healthy volunteers and patients with hypertension and angina. Subjects received doses of 120, 180, 360, or 540 mg racemic verapamil as an osmotic controlled-release formulation. A population pharmacokinetic model involving zero-order release of verapamil into the gastrointestinal tract with first-order absorption and elimination was used to describe the steady-state plasma concentration profile for R- and S-verapamil. A population sigmoid E(max) pharmacodynamic model was used to describe the effect of R- and S-verapamil on mean arterial blood pressure. RESULTS: S-verapamil had an approximate 4-fold greater apparent clearance than R-verapamil in both healthy volunteers and patients. The apparent plasma clearance of R- and S-verapamil in healthy volunteers decreased over the dose range of 120-540 mg. A similar dose-dependent decrease in apparent plasma clearance was also noted in patients. None of the patient demographic variables examined (age, total body weight, lean body weight, body mass index, and height) explained the variability in verapamil pharmacokinetics. The pharmacodynamic model describing the relationship between verapamil plasma concentration and mean arterial blood pressure indicated that the S-verapamil had a 3.6-fold lower estimated EC(50) compared to R-verapamil. CONCLUSIONS: The results from this dose ranging pharmacokinetic investigation in healthy volunteers and patients are consistent with previous reports in healthy subjects. S-verapamil is cleared more rapidly than R-verapamil and the estimated EC(50) for S-verapamil was 3.6-fold lower than for R-verapamil. Estimated EC(50) values for R- and S-verapamil decreased with increasing age and decreasing weight. PMID- 11891671 TI - Clinical pharmacokinetic studies with INN 00835 (nemifitide), a novel pentapeptide antidepressant. AB - Nemifitide (4-fluoro-L-phenylalanyl-trans-4-hydroxy-L-prolyl-L-arginylglycyl-L tryptophanamide ditrifluoroacetate) is a novel antidepressant, currently in phase 2/3 clinical trials. The purpose of our phase 1 clinical trials (conducted over a three year period) was to provide safety and pharmacokinetic data to support its clinical development as an antidepressant drug. Single and multiple doses ranging from 18 to 320 mg were administered subcutaneously to healthy volunteers in five phase 1 studies. Plasma concentrations of unchanged parent drug were determined by a validated LC/MS/MS method in blood samples collected at timepoints between 10 min and 72 h after dosing. Nemifitide was rapidly absorbed (C(max) at 10 min) and eliminated (t(1/2) 15-30 min) in most subjects. Regression and power model analyses were used to evaluate the data. The results indicate that pharmacokinetic parameters: AUC(0-t), AUC (0-infinity) and C(max), were close to dose proportional in the dose range investigated. There was no evidence of systemic accumulation of drug following 5 daily doses. No serious adverse events or clinically significant systemic adverse events occurred at any of the doses investigated in the over 100 subjects dosed in these studies. Drug-related adverse events were limited to local and transient skin reactions (pain and/or erythema) at the injection site, especially at the high doses administered: 240 and 320 mg. PMID- 11891672 TI - Disposition of radiolabeled BMS-204352 in rats and dogs. AB - BMS-204352, a maxi-K channel opener, is currently under development for the treatment of stroke. The objective of this study was to determine the pharmacokinetics, mass balance and absolute oral bioavailability of [(14)C]-BMS 204352 in rats and dogs. [(14)C]-BMS-204352 was administered, to rats (n=10/group; parallel design, 6 mg/kg) and dogs (n=4/group; crossover design, 2 mg/kg), as an oral (PO) or as a 3-min intraarterial (IA) infusion in rats and a 6 min intravenous (i.v.) infusion in dogs. Blood, urine, and feces samples were collected and analyzed for unchanged BMS-204352 (plasma) using a validated LC/MS assay and for total radioactivity (plasma, urine, feces) using liquid scintillation counting. The mean total body clearance (CLT) and steady-state volume of distribution (VSS) values for the unchanged BMS-204352 were 2.58 +/- 0.48 l/h/kg and 6.3 +/- 1.14 l/kg, respectively, in rats and 0.21 +/- 0.02 l/h/kg and 4.06 +/- 0.47 l/kg, respectively, in dogs. In both species, the elimination half-life of total radioactivity was significantly longer as compared to the unchanged drug. After IA administration of radiolabeled BMS-204352 to rats, ca. 5.9 and 85% of radioactivity was recovered within 7 days in urine and feces, respectively; corresponding recoveries after PO dosing were 4.5 and 99.5%, respectively. The recoveries were similar in dogs, i.e., ca. 5.2 and 83% of administered radioactivity recovered in urine and feces, respectively, for IV dose and ca. 4 and 86%, respectively, for PO dose. These data indicate that nonrenal (biliary) elimination in both species was predominant. Based on comparable urinary recovery of radioactivity and plasma AUCs of radioactivity, the extent of oral absorption of BMS-204352 appeared to be complete in both species. The absolute oral bioavailability was 55% in rats and 79% in dogs. Bioavailability and extent of absorption data suggest evidence of first pass metabolism of BMS-204352 in the rat and dog. PMID- 11891674 TI - Pseudoachondroplasia and multiple epiphyseal dysplasia: New etiologic developments. AB - Pseudoachondroplasia (PSACH) (OMIM#177170) and multiple epiphyseal dysplasia (MED) are separate but overlapping osteochondrodysplasias. PSACH is a dominantly inherited disorder characterized by short-limb short stature, loose joints, and early-onset osteoarthropathy. The diagnosis is based on characteristic clinical and radiographic findings. Only mutations in the cartilage oligomeric matrix protein (COMP) gene have been reported in PSACH, and all family studies have been consistent with linkage to the COMP locus on chromosome 19. Multiple epiphyseal dysplasia (MED) is a relatively mild chondrodysplasia but like PSACH, MED causes early-onset joint degeneration, particularly of the large weight-bearing joints. Given the clinical similarity between PSACH and MED, it was not surprising that the first MED locus identified was the COMP gene (EDM1). Mutations causing MED have now been identified in five other genes (COL9A1, COL9A2, COL9A3, DTDST, and MATN3), making MED one of the most genetically heterogeneous disorders. This article reviews the clinical features of PSACH and MED, the known mutations, and the pathogenetic effect of COMP mutations on the cartilage extracellular matrix. PMID- 11891675 TI - Col2-GFP reporter mouse--a new tool to study skeletal development. AB - Transgenic mice were generated that harbor a Col2-GFP reporter that marks chondrocytes and their immediate precursors during skeletal development. Cells engaged in chondrogenesis were identified by conventional fluorescence microscopy and confocal optical sectioning within their native environments in live embryos and in thick tissue slices. The use of these mice offers a novel approach for studying the role of chondrocytes in skeletal development. PMID- 11891676 TI - Dyssegmental dysplasia, Silverman-Handmaker type: unexpected role of perlecan in cartilage development. AB - Dyssegmental dysplasia, Silverman-Handmaker type (DDSH), is a lethal autosomal recessive form of dwarfism with characteristic anisospondylic micromelia. The remarkable similarities in the radiographic, clinical, and chondroosseous morphology of DDSH patients to those of perlecan-null mice led to the identification of mutations in the perlecan gene (HSPG2) of DDSH. Perlecan, a large heparan sulfate proteoglycan, is expressed in various tissues and is a component of all basement membrane extracellular matrices. A chondrodysplasia phenotype caused by the loss of perlecan was unexpected, because cartilage does not have basement membranes. Insertion and splicing mutations in HSPG2 of DDSH were found that were predicted to create a premature termination codon. Immunostaining and biochemical analysis revealed that the mutant perlecan molecules were unstable and not secreted into the extracellular matrix. These results indicate that DDSH is caused by functional null mutations of HSPG2 and that perlecan is essential for cartilage development. Published 2002 Wiley-Liss, Inc. PMID- 11891677 TI - Transcriptional dysregulation in skeletal malformation syndromes. AB - Normal skeletal development requires coordinated temporal and spatial gene expression patterns that specify the functions of various cell types. Transcription factors by definition coordinate this process and are themselves subject to hierarchical levels of regulation. Together they determine the context dependent function of each transcription factor. Hence, loss-of-function and gain of-function mutations within specific transcription factors cause dysregulation of broad transcriptional networks. Consequences are usually dominantly inherited skeletal malformation syndromes that can be broadly viewed as consequences of defects of cellular differentiation, proliferation, and survival versus defects in pattern formation. The study of human phenotypes and mutations can lead to hypotheses about targets within the respective transcriptional network. These targets can then be confirmed by combining mouse genetic and in vitro studies. Although this has been successful in a small group of skeletal dysplasias, the majority of transcriptional networks during skeletogenesis remain to be elucidated. PMID- 11891678 TI - Allelic and nonallelic heterogeneity in dyschondrosteosis (Leri-Weill syndrome). AB - Dyschondrosteosis (DCS) is an autosomal dominant form of mesomelic dysplasia that has been recently ascribed to large-scale deletions and nonsense mutations of the SHOX gene on the pseudoautosomal region of chromosome X and Y [Belin et al., 1998: Nat Genet 19:67-69; Shears et al., 1998: Nat Genet 19:70-73]. Here, we report the molecular analysis of a total of 23 DCS families including 16 previously reported pedigrees [Belin et al., 1998: Nat Genet 19:67-69; Huber et al., 2001: J Med Genet 38:281-284] and 7 novel DCS families. Linkage analyses in 21 of 23 families were consistent with linkage to the pseudoautosomal region. However, in 2 of 23 families, linkage studies excluded SHOX as the disease causing gene, suggesting that this condition is genetically heterogeneous. PMID- 11891679 TI - Comprehensive resource: Skeletal gene database. AB - The Skeletal Gene Database (SGD) is an integrated resource that provides comprehensive information about bone-related genes, mRNA, and proteins expressed in human and mouse, with rich links to numerous other electronic tools. SGD contains expressed sequence tag (EST) data from all the skeletal-related cDNA libraries that are available to the public. It supplies the query/data access analytic tools for users to search and compare each gene expressed in skeletal tissue(s). The results derived from EST tissue expression profiling will allow users to get the data on the mRNA copy numbers of each gene expressed in each tissue and its normalized value. From the SGD, researchers can obtain information regarding the name, symbol, size, exon/intron number, chromosomal location, LocusLink, and related disease (if any is known) of each gene. This electronic compendium also furnishes information on the protein of the corresponding gene including the protein size (amino acid number and molecular weight). It provides swift and ready access to other useful databases including OMIM, UniGene and PUBMED. The data will be updated regularly in step with current and future research, thereby providing what we hope will serve as a highly useful source of information and a powerful analytic tool to the scientific community. PMID- 11891680 TI - Molecular-pathogenetic classification of genetic disorders of the skeleton. AB - Genetic disorders of the skeleton (skeletal dysplasias and dysostoses) are a large and disparate group of diseases whose unifying features are malformation, disproportionate growth, and deformation of the skeleton or of individual bones or groups of bones. To cope with the large number of different disorders, the "Nosology and Classification of the Osteochondrodysplasias," based on clinical and radiographic features, has been designed and revised periodically. Biochemical and molecular features have been partially implemented in the Nosology, but the rapid accumulation of knowledge on genes and proteins cannot be easily merged into the clinical-radiographic classification. We present here, as a complement to the existing Nosology, a classification of genetic disorders of the skeleton based on the structure and function of the causative genes and proteins. This molecular-pathogenetic classification should be helpful in recognizing metabolic and signaling pathways relevant to skeletal development, in pointing out candidate genes and possible therapeutic targets, and more generally in bringing the clinic closer to the basic science laboratory and in promoting research in this field. PMID- 11891681 TI - "Mowat-Wilson" syndrome with and without Hirschsprung disease is a distinct, recognizable multiple congenital anomalies-mental retardation syndrome caused by mutations in the zinc finger homeo box 1B gene. AB - Recently mutations in the gene ZFHX1B (SIP1) were shown in patients with "syndromic Hirschsprung disease" with mental retardation (MR) and multiple congenital anomalies (MCA), but it was unclear if Hirschsprung disease is an obligate symptom of these mutations and if the distinct facial phenotype delineated by Mowat et al. [1998: J Med Genet 35: 617-623] is specific for ZFHX1B mutations. In order to address these open questions we analyzed the ZFHX1B gene in five patients, three of whom had "syndromic Hirschsprung disease" two with and one without the facial phenotype described by Mowat et al. [1998], and two of whom had the distinct facial gestalt without Hirschsprung disease. Analyses of microsatellite markers and newly identified SNPs, and/or FISH with BACs from the ZFHX1B region excluded large deletions in all five patients. Direct sequencing demonstrated truncating ZFHX1B mutations in all four patients with the characteristic facial phenotype, but not in the patient with syndromic Hirschsprung disease without the distinct facial appearance. We demonstrate that there is a specific clinical entity with a recognizable facial gestalt, mental retardation and variable MCAs which we propose be called the "Mowat-Wilson syndrome." PMID- 11891682 TI - Identification of de novo chromosome rearrangements: five cases analyzed with differential chromosome painting. AB - We report on five cases of de novo structural chromosome rearrangements that were difficult to identify by conventional G-banding analysis. In all five cases, differential chromosome painting (DCP) provided evidence for the presence of an additional segment and its origin. A combination of DCP with subsequent conventional fluorescence in situ hybridization (FISH) analysis using adequate locus-specific probes and reexamination of G-banding patterns resulted in successful identification of the rearrangements. Their karyotypes were finally interpreted as 46,XY,der(1)(qter --> q42.1::p36.3 --> qter) in case 1; 46,XY,der(8)(8pter -->8q24.3::8q24.3 --> 8q23.2::?p11.2 --> ?ps) in case 2; 47,XY,+der(10)(pter --> q11) in case 3; 46,XX,der(3)(17pter --> 17p11.2::3p26 --> 3qter) in case 4; and 46,XY,dup(1) (pter --> q32::q25 --> qter) in case 5. PMID- 11891683 TI - Heterogeneity in familial dominant Paget disease of bone and muscular dystrophy. AB - The combination of autosomal dominant, early onset Paget disease of bone (PDB) and muscular dystrophy is an unusual disorder. We recently mapped the disorder in a large family from central Illinois with PDB and proximal limb-girdle type of muscular dystrophy (LGMD), and in 3 additional families with hereditary inclusion body myopathy (HIBM), Paget disease of bone and frontotemporal dementia, to a unique locus on chromosome 9p21.1-q12. The present study describes an unrelated 10-member family with autosomal dominant PDB and a scapuloperoneal type of muscular dystrophy. Clinical, biochemical, and radiological evaluations were performed to delineate clinical features in this family. Progression of the muscular dystrophy begins with weakness in the distal muscles of the legs accompanied by foot drop. EMG and muscle biopsy are compatible with a primary dystrophy. Onset of Paget disease is early, at a mean age of 41 years, with initial distribution in the long bones and eventual infiltration of the spine and pelvis. Creatine phosphokinase (CPK) and alkaline phosphatase levels are elevated in affected individuals. Molecular analyses excluded all known loci for Paget disease of bone, scapuloperoneal muscular dystrophy (SPMD), fascioscapulohumeral muscular dystrophy (FSH), amyotrophic lateral sclerosis (ALS), Bethlem myopathy, two forms of autosomal dominant limb-girdle muscular dystrophy (LGMD), and the critical region for LGMD or HIBM/PDB on chromosome 9p21.1-q12, thus providing evidence for genetic heterogeneity among families with the unique combination of muscular dystrophy and Paget disease of bone. PMID- 11891684 TI - Delineation of the dup5q phenotype by molecular cytogenetic analysis in a patient with dup5q/del 5p (cri du chat). AB - An infant girl presented with multiple congenital abnormalities and a distinctive mewing cry. Her karyotype was 46,XX,add5p. Chromosome analysis on the mother revealed an apparently balanced pericentric inversion of chromosome 5, with the precise position of the breakpoints not clearly discernable by GTG banding, 46,XX,inv(5)(p15.2/3?q35.1?). Fluorescence in situ hybridization (FISH) studies using a commercial cri du chat probe (D5S721,D5S23) revealed signals on both the normal and derivative chromosomes. Telomeric probes specific for 5p and 5q were used to confirm the pericentric inversion in the mother and demonstrated the loss of the terminal 5p region and a duplication of the terminal 5q region in the proband. The imbalance on chromosome 5 in the patient was further defined using comparative genomic hybridization (CGH), which revealed a loss of material from 5p15.3 --> pter and a gain of 5q34 --> qter. The presence of the cat-like cry appears to be the only specific feature that can be linked to the loss of 5p material. The remaining dysmorphic features of this infant appear to be due specifically to the duplication of the 5q sequences. The combination of FISH, CGH, and cytogenetics has confirmed that the characteristic cry of the cri du chat syndrome is due to the deletion of the most distal part of the classic del 5p region. More importantly, our investigation has defined the duplication of 5q34 --> qter as a distinct clinical phenotype. PMID- 11891685 TI - Unique case of mosaicism involving two morphologically similar marker chromosomes of different centric origin in a patient with developmental delay. AB - A five-year-old Caucasian male presented with developmental delay, minor dysmorphic features, and hyperactivity. Cytogenetic analysis showed the presence of a marker chromosome in the majority of cells analyzed. Fluorescence in situ hybridization (FISH) analyses using several alpha satellite probes, including D13Z1/D21Z1, did not reveal any signal on the marker chromosome. Subsequent multicolor FISH (M-FISH) indicated the marker to be derived from chromosome 13, and FISH with a chromosome 13 paint confirmed this finding. The absence of D13Z1/D21Z1 signal on the marker suggested that it was analphoid in nature. Comparative genomic hybridization (CGH) was utilized to further characterize the region of chromosome 13 from which the marker originated, and unexpectedly revealed a gain of chromosomal material at both the centromeric regions of chromosomes 3 and 13. In view of the CGH results, extensive FISH studies with D3Z1 and D13Z1/D21Z1 were performed and revealed the presence of four cell lines comprising one normal cell line (50.5%), a cell line with a chromosome 3 derived marker (19%), a cell line containing a marker derived from chromosome 13 (20%), and a cell line with both markers (10.5%). As the two markers appeared morphologically similar by GTG banding, all 47,XY metaphases in the initial analysis were thought to comprise only a single marker. This is the first report, to our knowledge, of the presence of a chromosome 3 and a chromosome 13 marker in mosaic condition in a congenital disorder. In light of our experience, we urge caution in interpreting karyotypes with marker chromosomes. Our case illustrates the limitations of fluorescent DNA probes and sampling errors. PMID- 11891686 TI - A combination of physical examination and ECG detects the majority of hemodynamically significant heart defects in neonates with Down syndrome. AB - Echocardiography has become the method of choice in the diagnosis of a congenital heart defect (CHD) in neonates with Down syndrome. The most compelling argument for diagnosis of CHD in the neonatal period is the need for early surgical intervention (ideally prior to 6 months of age) in those with a complete atrioventricular (AV) canal. We evaluated the efficacy of a combined approach of physical examination (PE) and electrocardiography (ECG) in the detection of CHD in 49 neonates with Down syndrome. Our findings indicate that most hemodynamically significant defects (78%), especially a complete AV canal, can be detected by this approach. Hemodynamically insignificant minor defects, such as a small patent ductus arteriosus (PDA) and a small atrial septal defect (ASD), may be missed. Thus, echocardiography remains the most sensitive way to diagnose CHD. However, given that the combination of PE and ECG detects the majority of complete AV canal defects, it can be used as an alternative approach when echocardiography is not easily accessible, due to geographic or economic constraints. Follow-up after a few weeks of those with normal PE and ECG findings should enable detection of new symptoms and signs and evolving ECG findings that may have been missed in the immediate newborn period. Patients who are judged by PE and ECG to have CHD will need echocardiographic confirmation. PMID- 11891687 TI - Absence of thumbs, A/hypoplasia of radius, hypoplasia of ulnae, retarded bone age, short stature, microcephaly, hypoplastic genitalia, and mental retardation. AB - The previously unreported combination of bilateral absence of thumbs, aplasia of ulna at one and hypoplasia of ulna on the other side, retarded bone age, short stature, microcephaly, micropenis, cryptorchidism, and mental retardation is described in a 5-year-old boy. Having excluded major differential diagnoses, e.g. Fanconi anemia, RAPADILINO syndrome and VACTERL association, we hypothesize that this boy represents a new multiple congenital anomaly/mental retardation (MCA/MR) syndrome. PMID- 11891688 TI - Clinical variability in mucolipidosis III (pseudo-Hurler polydystrophy). AB - Mucolipidosis III (MLIII) is caused by a deficiency of UDP-N acetylglucosamine:lysosomal enzyme N-acetylglucosamine 1-phosphotransferase (phosphotransferase) activity, an enzyme responsible for the formation of the recognition marker on most lysosomal enzymes. The consequences of this defect are impairment of many lysosomal catabolic processes. A deficiency of phosphotransferase activity causes two phenotypically different diseases: mucolipidosis II and a rare form, mucolipidosis III (pseudo-Hurler polydystrophy). The purpose of this article is to report three patients with ML III, presenting quite different clinical courses: Patient 1 is a 13-year-old girl in whom the only symptoms of ML III were joint stiffness of the hands. Patients 2 and 3 are sibs: a 5-year-old boy with a severe form of ML III and his 2-year-old sister, who is less affected than her brother at the same age. A comparison of biochemical results and the clinical picture of our patients with cases in the literature is presented. PMID- 11891689 TI - Childhood hyperuricemia and acute renal failure resulting from a missense mutation in the HPRT gene. AB - A 6-year-old boy was determined to have partial hypoxanthine phosphoribosyl transferase (HPRT) enzyme deficiency without the phenotypic features of Lesch Nyhan syndrome. He presented with recurrent acute renal failure (ARF) from hyperuricemia. Treatment with allopurinol prevented further attacks of renal failure. T lymphocyte cultures were used to sequence the HPRT cDNA and a novel single nucleotide substitution at codon 65 in exon 3 was found (193C>T, 65leu>phe). This mutation was confirmed by genomic DNA sequencing and was also detected in his heterozygous, asymptomatic mother and sister. Unlike the cells from patients with classic Lesch-Nyhan syndrome, the in vitro cultures of our patient's T-lymphocytes did not proliferate in the presence of purine analogue 6 thioguanine (TG). This report highlights the unusual occurrence of recurrent ARF in a child with partial HPRT enzyme deficiency. The absence of TG resistance in vitro with this mutation shows that even small alterations in enzyme activity in vivo can result in disease symptoms, in this instance, hyperuricemia sufficient to cause ARF. Atypical HPRT mutations should also be considered in cases of unusual renal failure, because correct diagnosis can allow appropriate treatment, as well as informed genetic counseling. PMID- 11891690 TI - ABCD syndrome is caused by a homozygous mutation in the EDNRB gene. AB - ABCD syndrome is an autosomal recessive syndrome characterized by albinism, black lock, cell migration disorder of the neurocytes of the gut (Hirschsprung disease [HSCR]), and deafness. This phenotype clearly overlaps with the features of the Shah-Waardenburg syndrome, comprising sensorineural deafness; hypopigmentation of skin, hair, and irides; and HSCR. Therefore, we screened DNA of the index patient of the ABCD syndrome family for mutations in the endothelin B receptor (EDNRB) gene, a gene known to be involved in Shah-Waardenburg syndrome. A homozygous nonsense mutation in exon 3 (R201X) of the EDNRB gene was found. We therefore suggest that ABCD syndrome is not a separate entity, but an expression of Shah Waardenburg syndrome. PMID- 11891691 TI - Patient with rheumatoid arthritis and MCA/MR syndrome due to unbalanced der(18) transmission of a paternal translocation t(18;20)(p11.1;p11.1). AB - A girl with psychomotor retardation and a pattern of minor anomalies was found to have a slightly enlarged short arm of chromosome 18 by conventional GTG-banded chromosome examination. The 18p+chromosome has also been found in the father. FISH studies using chromosome 18-and chromosome 20-specific painting probes confirmed a reciprocal whole arm translocation between chromosomes 18 and 20 in the father, resulting in monosomy of the short arm of chromosome 18 and trisomy of the short arm chromosome 20 in the patient. FISH analysis using a chromosome 18 alpha-satellite-specific probe showed a reduced signal intensity. The patient presented with a flat, oval face, upslanting palpebral fissures, periorbital fullness, and mental retardation; she also had chronic diarrhea with milk protein intolerance and juvenile rheumatoid arthritis at age 5 years. Juvenile rheumatoid arthritis, like several other immunologic disorders, has occasionally been reported in patients with deletion of 18p, and thus most likely loss of a gene or genes on 18p is responsible for various immunologic disorders occurring in these patients. PMID- 11891693 TI - Familial digital arthropathy-brachydactyly. AB - We report a large family with a previously undescribed, dominantly inherited condition comprising arthropathy of the hands and feet and progressive shortening of the middle and distal phalanges. We have designated the condition familial digital arthropathy-brachydactyly (FDAB). Onset of FDAB is in the first decade and the arthropathy is progressive, resulting in deformity and pain in adult life. The remainder of the skeleton is not affected. It is hypothesized from the radiological appearance of patients at different ages that FDAB might result from subchondral pathology primarily affecting the heads of the phalanges, metacarpals, and metatarsals, with the arthropathy and brachydactyly being secondary effects. PMID- 11891692 TI - PAGOD syndrome: eighth case and comparison to animal models of congenital vitamin A deficiency. AB - We observed a 46, XY infant with atrophy of the optic nerve, complex congenital heart disease including a double outlet right ventricle, hypoplasia of the right pulmonary artery and lung, eventration of the diaphragm, and ambiguous genitalia. The baby died of cardiac arrhythmias at 204 days. The pattern of malformations was compatible with pulmonary tract and pulmonary artery, agonadism, omphalocele, diaphragmatic defect, and dextrocardia (PAGOD) syndrome. The condition may resemble the malformation complex associated with developmental deficiency of vitamin A or retinoic acid, as described in animal models. PMID- 11891694 TI - A deletion-insertion mutation in the phosphomannomutase 2 gene in an African American patient with congenital disorders of glycosylation-Ia. AB - Congenital disorders of glycosylation (CDG) are a group of metabolic disorders with multisystemic involvement characterized by abnormalities in the synthesis of N-linked oligosaccharides. The most common form, CDG-Ia, resulting from mutations in the gene encoding the enzyme phosphomannomutase (PMM2), manifests with severe abnormalities in psychomotor development, dysmorphic features and visceral involvement. While this disorder is panethnic, we present the first cases of CDG Ia identified in an African American family with two affected sisters. The proband had failure to thrive in infancy, hypotonia, ataxia, cerebellar hypoplasia and developmental delay. On examination, she also exhibited strabismus, inverted nipples and an atypical perineal fat distribution, all features characteristic of CDG-Ia. Direct sequencing demonstrated that the patient had a unique genotype, T237M/c.565-571 delAGAGAT insGTGGATTTCC. The novel deletion-insertion mutation, which was confirmed by subcloning and sequencing of each allele, introduces a stop codon 11 amino acids downstream from the site of the deletion. The presence of this deletion-insertion mutation at cDNA position 565 suggests that this site in the PMM2 gene may be a hotspot for chromosomal breakage. PMID- 11891695 TI - Septo-optic dysplasia and digital anomalies: another observation. PMID- 11891696 TI - Frontometaphyseal dysplasia: patient with ruptured aneurysm of the aortic sinus of Valsalva and cerebral aneurysms. PMID- 11891697 TI - Psoriasis vulgaris in a male with partial deletion 18p. PMID- 11891698 TI - Nowadays it is preceptive to perform chromosomal studies with high resolution G bands and FISH techniques when necessary. PMID- 11891703 TI - On-chip enzymatic assays. AB - This article reviews different possibilities for conducting enzymatic assays on microchip platforms, along with potential advantages, limitations, and selected examples of such biochips. Enzyme-based chips combine the analytical power and reagent economy of microfluidic devices with the selectivity and amplification features of biocatalytic reactions. "Lab-on-chip" devices thus allow enzymatic assays to be performed more rapidly, easily, and economically. Such assays usually rely on on-chip mixing and reactions (of the substrates and enzymes) in connection to separations (of the substrates or products). The realization of on chip enzymatic assays thus requires understanding of how enzymatic reactions behave on a small scale and can be interfaced with separation microchips, and how the microfluidics can be tailored to suit the requirements of particular enzymatic assays. The goal is to obtain sufficient reaction times, without compromising the quality of the analytical separation. The versatility of such on chip enzymatic assays offers great promise for decentralized testing of clinically or environmentally important substrates. PMID- 11891702 TI - Microfluidic chips for clinical and forensic analysis. AB - This review gives an overview of developments in the field of microchip analysis for clinical diagnostic and forensic applications. The approach chosen to review the literature is different from that in most microchip reviews to date, in that the information is presented in terms of analytes tested rather than microchip method. Analyte categories for which examples are presented include (i) drugs (quality control, seizures) and explosives residues, (ii) drugs and endogenous small molecules and ions in biofluids, (iii) proteins and peptides, and (iv) analysis of nucleic acids and oligonucleotides. Few cases of microchip analysis of physiological samples or other "real-world" matrices were found. However, many of the examples presented have potential application for these samples, especially with ongoing parallel developments involving integration of sample pretreatment onto chips and the use of fluid propulsion mechanisms other than electrokinetic pumping. PMID- 11891704 TI - High-speed analysis of multiplexed short tandem repeats with an electrophoretic microdevice. AB - We report the development of a robust and effective method for multiplexed short tandem repeat (STR) analysis within a chip-based microdevice. The method uses a laser-induced fluorescence detection system and simultaneously detects three- and four-color multiplexed polymerase chain reaction (PCR) samples. Analyses of the eight combined DNA index system (CODIS) STR loci were performed in 20 min with single-base-pair resolution ranging from 0.75 to 1. A simultaneous analysis of fifteen loci-ladders and a gender marker Amelogenin based on the PowerPlexTM 16 System was achieved in less than 35 min. The system is capable of repetitive operation and may be extended to high-throughput multilane devices that could be readily interfaced to an automated sample loading system. PMID- 11891705 TI - Toward a microchip-based solid-phase extraction method for isolation of nucleic acids. AB - A silica-based solid-phase extraction system suitable for incorporation into a microchip platform (nu-total analytical system; nu-TAS) would find utility in a variety of genetic analysis protocols, including DNA sequencing. The extraction procedure utilized is based on adsorption of the DNA onto bare silica. The procedure involves three steps: (i) DNA adsorption in the presence of a chaotropic salt, (ii) removal of contaminants with an alcohol/water solution, and (iii) elution of the adsorbed DNA in a small volume of buffer suitable for polymerase chain reaction (PCR) amplification. Multiple approaches for incorporation of this protocol into a microchip were examined with regard to extraction efficiency, reproducibility, stability, and the potential to provide PCR-amplifiable DNA. These included packing microchannels with silica beads only, generating a continuous silica network via sol-gel chemistry, and combinations of these. The optimal approach was found to involve immobilizing silica beads packed into the channel using a sol-gel network. This method allowed for successful extraction and elution of nanogram quantities of DNA in less than 25 min, with the DNA obtained in the elution buffer fraction. Evaluation of the eluted DNA indicated that it was of suitable quality for subsequent amplification by PCR. PMID- 11891706 TI - Microchip-based immunoassay system with branching multichannels for simultaneous determination of interferon-gamma. AB - A bead-bed immunoassay system suitable for simultaneous assay of multiple samples was constructed on a microchip. The chip had branching multichannels and four reaction and detection regions; the constructed system could process four samples at a time with only one pump unit. Interferon gamma was assayed by a 3-step sandwich immunoassay with the system coupled to a thermal lens microscope as a detector. The biases of the signal intensities obtained from each channel were within 10%, and coefficients of variation were almost the same level as the single straight channel assay. The assay time for four samples was 50 min instead of 35 min for one sample in the single-channel assay; hence higher throughput was realized with the branching structure chip. PMID- 11891707 TI - Application of surface biopassivated disposable poly(dimethylsiloxane)/glass chips to a heterogeneous competitive human serum immunoglobulin G immunoassay with incorporated internal standard. AB - A microfluidic platform for a heterogeneous competitive immunoassay of human immunoglobulin G (IgG) employing Cy5-human IgG as tracer and Cy3-mouse IgG as internal standard was developed. The device consisted of microchannels made of poly(dimethylsiloxane) and glass which were patterned with antibodies against human IgG and mouse IgG. Electrokinetic sample transport was employed in order to exploit the small difference between the net mobilities of analyte and tracer, thereby achieving favorable conditions for the performance of the competitive immunoreaction. The overall quality of the disposable chip and performance of the immunoassay were controlled by monitoring the fluorescence of bound tracer and bound internal standard. Analyses with an insufficient internal standard response were discarded, and immunoassay data evaluation was based on the ratio of tracer and internal standard fluorescence. Using synthetic samples in the range from 0 to 80 microg/mL IgG and alkaline running conditions, a concentration-dependent response with reproducible Cy5/Cy3 signal ratios (average relative standard deviation of 6.8%) was obtained. Chips stored with solution in the channels at 4 degrees C over a two-month period were found to perform like freshly prepared chips, whereas chips stored dry at -20 degrees C and rehydrated prior to use could not be employed. The analysis of patient sera showed that the immunoassay platform behaved differently in the presence of serum-based samples. Using the same conditions as for the synthetic samples, no concentration dependence was noted. With a large excess of tracer, however, an IgG concentration dependence was observed, permitting distinction of samples of patients with normal IgG serum levels (8-16 mg/mL) from those with elevated IgG concentrations (>16 mg/mL). PMID- 11891708 TI - Affinity protection chromatography for efficient labeling of antibodies for use in affinity capillary electrophoresis. AB - Capillary electrophoresis immunoassay (CEIA) is shown to be substantially more sensitive to the antibody (Ab) reagent quality than are immunosorbent methods such as enzyme-linked immunosorbent assays (ELISA). Cyanine 5 (Cy5)-labeled monoclonal anti-ovalbumin (mAb*) was inactive for CEIA of ovalbumin (Ov), yet was functional in ELISA for Ov. ELISA showed the mAb* was at least ten times less active, accounting for the poor CEIA performance. Labeled polyclonal Ab was inactive for a dye to protein ratio greater than 1.6. An affinity protection chromatography procedure (APC) was developed for Ab labeling, which avoided degradation of the Ab binding site. Ov was covalently bound to cyanogen bromide activated cellulose gel in a column, and used to capture the Ab. The coupling efficiency for Ov to the gel was 74-97%, Ab could then be bound with 95-100% efficiency, and Ab* was recovered in 50% yield following labeling on the column. This procedure was performed successfully in three different laboratories, indicating the robustness of the optimized APC synthetic method. No inactive Ab* could be detected in the APC product. The CEIA detection limit for ovalbumin using APC labeled mAb was 173 nM, when [Ab*] was fixed at 163 nM. The association constants of mAb and mAb* were determined by CEIA. PMID- 11891709 TI - Detection of homocysteine by conventional and microchip capillary electrophoresis/electrochemistry. AB - A method based on capillary electrophoresis (CE) with electrochemical (EC) detection for the determination of both total homocysteine (tHcy) and protein bound homocysteine (pbHcy) in plasma is described. Both end-column and off-column amperometric detection were investigated. Off-column detection resulted in a more sensitive assay for the determination of homocysteine (Hcy). The detection limit for homocysteine was 500 nM using off-column EC detection and the response was linear over the range 1-100 microM. Therefore, this assay is appropriate for the quantification of Hcy over the physiological concentration ranges found in all disease states. Methodologies for the determination of tHcy and pbHcy in human plasma were investigated and optimized and the concentrations of both pbHcy and tHcy in plasma obtained from a healthy individual were determined to be 2.79+/ 0.31 nuM (n = 4) and 3.37+/-0.15 microM (n = 3), respectively. The methodology was then transferred to a microchip CE-EC format and Hcy and reduced glutathione (GSH) were detected. Future work will focus on the development of ancillary methodologies to identify the other forms of Hcy in vivo. PMID- 11891710 TI - The analysis of uric acid in urine using microchip capillary electrophoresis with electrochemical detection. AB - Clinical studies have linked irregular concentrations of uric acid in urine to several diseases. Conventional methods for the measurement of uric acid are however temperature-dependent, expensive, and require labile reagents. The miniaturization of analytical techniques, specifically capillary electrophoresis, offers an ideal alternative for clinical analyses such as uric acid determination. The added benefits include reduced reagent and analyte consumption, decreased maintenance costs, and increased throughput and portability. A microchip capillary electrophoresis-electrochemical system for the analysis of uric acid in urine is described. The poly(dimethylsiloxane) (PDMS)/glass microchip utilizes amperometric detection via an off-chip platinum working electrode. Linear responses from 1 to 165 microM and 15 to 110 microM were found for dopamine and uric acid, respectively. The limit of detection for both compounds was 1 microM. Once characterized, the system was used to measure the concentration of uric acid in a dilute urine sample in less than 30 s. The measured uric acid concentration was verified with the uricase reaction and found to be acceptable. Six additional urine samples were evaluated with the microchip device and the uric acid concentration for each sample was found to be in the expected clinical concentration range. PMID- 11891711 TI - Determination of oxalate in urine by zone electrophoresis on a chip with conductivity detection. AB - The use of a poly(methylmethacrylate) capillary electrophoresis chip, provided with a high sample load capacity separation system (a 8500 nL separation channel coupled to a 500 nL sample injection channel) and a pair of on-chip conductivity detectors, for zone electrophoresis (ZE) determination of oxalate in urine was studied. Hydrodynamic and electroosmotic flows of the solution in the separation compartment of the chip were suppressed and electrophoresis was a dominant transport process in the separations performed on the chip. A low pH of the carrier electrolyte (4.0) provided an adequate selectivity in the separation of oxalate from anionic urine constituents and, at the same time, also a sufficient sensitivity in its conductivity detection. Under our working conditions, this anion could be detected at a 8 x 10(-8) mol/L concentration also in samples containing chloride (a major anionic constituent of urine) at 3.5 x 10(-3) mol/L concentrations. Such a favorable analyte/matrix concentration ratio (in part, attributable to a transient isotachophoresis stacking in the initial phase of the separation) made possible accurate and reproducible (typically, 2-5% relative standard deviation (RSD) values of the peak areas of the analyte in dependence on its concentration in the sample) determination of oxalate in 500 nL volumes of 20 100-fold diluted urine samples. Short analysis times (about 280 s), no sample pretreatment (not considering urine dilution) and reproducible migration times of this analyte (0.5-1.0% RSD values) were characteristic for ZE on the chip. This work indicates general potentialities of the present chip design in rapid ZE analysis of samples containing the analyte(s) at high ionic matrix/analyte concentration ratios. PMID- 11891712 TI - Polymer microchips bonded by O2-plasma activation. AB - This paper presents a fabrication of polymer microchips with homogeneous material technique due to surface treatment by plasma before sealing. UV laser photoablation was used for fast prototyping of microstructures, and oxygen plasma was used as a surface treatment for both the microfabricated substrate and the polymer cover. It was found that with an oxidative plasma treatment, successful bonding could be achieved without adhesive material between polymer sheets substantially below the glass transition temperature of the polymer. Homogeneous polyethylene terephthalate (PET) microstructures were characterized by scanning electron microscopy (SEM) and analyzed by X-ray photoelectron spectroscopy (XPS) surface analyses after different surface treatments. The electroosmotic flow characteristics including the velocity and the stability over 20 days have been tested and compared to composite channels, in which the cover presents a polyethylene (PE) adhesive layer. Capillary zone electrophoresis in both homogeneous and composite microanalytical devices were then performed and compared in order to evaluate the separation efficiency. In preliminary experiments, a plate height of 0.6 microm has been obtained with homogenous microchannels. The surface analysis pointed out that the surface chemistry is of prime importance for the performance of microfluidic separation. PMID- 11891713 TI - Control of electroosmotic flow in laser-ablated and chemically modified hot imprinted poly(ethylene terephthalate glycol) microchannels. AB - The fabrication of microchannels in poly(ethylene terephthalate glycol) (PETG) by laser ablation and the hot imprinting method is described. In addition, hot imprinted microchannels were hydrolyzed to yield additional charged organic functional groups on the imprinted surface. The charged groups are carboxylate moieties that were also used as a means for the further reaction of different chemical species on the surface of the PETG microchannels. The microchannels were characterized by fluorescence mapping and electroosmotic flow (EOF) measurements. Experimental results demonstrated that different fabrication and channel treatment protocols resulted in different EOF rates. Laser-ablated channels had similar EOF rates (5.3+/-0.3 x 10(-4) cm(2)/Vs and 5.6+/-0.4 x 10(-4) cm(2)/Vs) to hydrolyzed imprinted channels (5.1+/-0.4 x 10(-4) cm(2)/Vs), which in turn demonstrated a somewhat higher flow rate than imprinted PETG channels that were not hydrolyzed (3.5+/-0.3 x 10(-4) cm(2)/Vs). Laser-ablated channels that had been chemically modified to yield amines displayed an EOF rate of 3.38+/- 0.1 x 10(-4) cm(2)/Vs and hydrolyzed imprinted channels that had been chemically derivatized to yield amines showed an EOF rate of 2.67+/-0.6 cm(2)/Vs. These data demonstrate that surface-bound carboxylate species can be used as a template for further chemical reactions in addition to changing the EOF mobility within microchannels. PMID- 11891714 TI - Liposomes as signal amplification reagents for bioassays in microfluidic channels. AB - Liposomes with encapsulated carboxyfluorescein were used in an affinity-based assay to provide signal amplification for small-volume fluorescence measurements. Microfluidic channels were fabricated by imprinting in a plastic substrate material, poly(ethylene terephthalate glycol) (PETG), using a silicon template imprinting tool. Streptavidin was linked to the surface through biotinylated protein for effective immobilization with minimal nonspecific adsorption of the liposome reagent. Lipids derivatized with biotin were incorporated into the liposome membrane to make the liposomes reactive for affinity assays. Specific binding of the liposomes to microchannel walls, dependence of binding on incubation time, and nonspecific adsorption of the liposome reagent were evaluated. The results of a competitive assay employing liposomes in the microchannels are presented. PMID- 11891715 TI - Brownian motion of single molecules in electric fields. AB - The behavior of negatively charged single rhodamine-labeled molecules excited to fluorescence in confocal volume elements has been investigated in oscillating voltage gradients between 1 and 15 kV/cm. The effective charge of rhodamin labeled deoxy-uridine triphosphate (Rh-dUTP) is dependent on the field strength applied and likely is due to differences in the screening by counter ions. It can be shown that the attractive forces generated by an oscillating dipole can increase the concentration of Rh-dUTP by a factor 2. Likewise a substantial increase of concentration can be obtained for Rh-labeled DNA molecules when linear voltage gradients are applied. The importance for diagnostic analysis at the single molecule level is discussed. PMID- 11891716 TI - A Fourier analysis approach for capillary polarimetry. AB - A new method of fringe interrogation based on Fourier analysis was implemented and tested for a capillary polarimetry detector. It has significant advantages over the previously employed depth of modulation (DOM) approach, including speed and alignment insensitivity. The new and old methods were compared using a set of interference fringes typically used to facilitate nanoliter volume polarimetric determinations. Polarimetric response was calculated with both methods over the range from 0 degrees to 180 degrees. The results were found to be in good agreement with Malus Law and indicate that an fast Fourier transform (fft) could be used for real-time capillary scale polarimetry in a probe volume of 40 nL. PMID- 11891718 TI - Kinetics of HL-60 cell entry to apoptosis during treatment with TNF-alpha or camptothecin assayed by the stathmo-apoptosis method. AB - BACKGROUND: Duration of apoptosis, from onset to final disintegration of the cell, is often short and variable. The apoptotic index (AI), as a snapshot of a transient event of variable length, does not truly represent incidence of apoptosis in the studied cell population. We recently proposed to estimate the cumulative apoptotic index (CAI) by inducing stathmo-apoptosis. A fluorescent inhibitor of caspases (FLICA) FAM-VAD-FMK is used to arrest the process of apoptosis and thereby prevent cell disintegration. Simultaneously, the arrested/apoptotic cells become FLICA-labeled. In the present study, this approach was applied to measure kinetics of HL-60 cell entrance into apoptosis induced via cell surface death receptor or a mitochondria-initiated pathway. Materials and Methods Cultures of HL-60 cells were treated with either TNF-alpha or camptothecin (CPT) in the absence or constant presence of 10-50 microM FLICA. The CAI was measured at different time points for up to 48 h by flow cytometry. Bivariate analysis of DNA content and cell labeling with FLICA was used to correlate apoptosis with the cell-cycle position. RESULTS: Selective loss of apoptotic cells seen in HL-60 cell cultures exposed to either TNF-alpha or CPT alone was prevented in cultures containing FLICA. Addition of FLICA alone had no effect on cell viability. The percentage of FLICA-labeled cells was plotted as a function of time after addition of TNF-alpha or CPT. The rate of cell entry to apoptosis was subsequently estimated from the slopes of the stathmo-apoptotic plot. The slopes revealed that the TNF-alpha or CPT-treated cells asynchronously underwent apoptosis with a stochastic-like kinetics and at two different rates. About 50% of cells in the TNF-alpha-treated cultures underwent apoptosis during the initial 6 h at a rate of approximately 8% of cells per hour; the remaining cells were undergoing apoptosis at a rate of approximately 2.5% of cells per hour for up to 24 h. Also, about 50% of the CPT-treated cells, predominantly those in S phase of the cell cycle, underwent apoptosis within the initial 8 h of CPT exposure, at a rate of approximately 7% of cells per hour. Remaining cells were undergoing apoptosis at a rate of approximately 1% of cells per hour during up to 48 h exposure to CPT. Spontaneous apoptosis in the untreated cultures occurred at a rate of 0.2% of cells per hour. CONCLUSIONS: This approach provides a means for measuring the kinetics of cell entrance to apoptosis (caspase activation) in large populations of cells in relation to the cell-cycle position. PMID- 11891719 TI - Phosphorylation at threonine-18 in addition to phosphorylation at serine-19 on myosin-II regulatory light chain is a mitosis-specific event. AB - BACKGROUND: Cell division is an inevitable and vitally indispensable event in cell life, when the nucleus and cytoskeleton undergo profound reorganization. Cytoplasmic division (cytokinesis) is known to occur immediately after the end of nuclear division, when the nuclear envelope breaks down, and chromosomes condense and segregate, but its driving mechanism remains enigmatic. Myosin, particularly myosin-II, is thought to be required for cytokinesis as a force-generating element, the activity of which is mainly regulated through phosphorylations on its 20-kDa regulatory light chains (RLCs). MATERIALS AND METHODS: Multiparameter flow cytometric analysis was performed on fixed HeLa S3 cells (suspension culture cells) sequentially stained with the polyclonal antibody (termed PP1) against both phosphorylated sites (serine-19 and threonine-18) on the RLC, and with propidium iodide for DNA. "Positive" cells were sorted, followed by their microscopic examination. Fluorescence microscopy was employed to visualize the cell-cycle-dependent distribution of immunolabeled diphosphorylated RLCs in both HeLa S3 and adherent HeLa cells. RESULTS AND CONCLUSIONS: Doubly phosphorylated myosin RLCs were highly expressed in mitotic cells, suggesting the positive regulatory role of diphosphorylation in the redistribution of RLCs between daughter cells and then in cytokinesis. The increased immunofluorescence signal from the phosphorylated forms of RLC, together with flow cytometry, provides a clue with which to investigate the mechanisms governing the function of nonmuscle myosins during various cell motile events, including cytokinesis. PMID- 11891720 TI - Flow cytometric cytotoxicity assay for measuring mammalian and avian NK cell activity. AB - BACKGROUND: Flow-cytometric assays are convenient alternatives to classic radioactive natural killer (NK) tests. MitoTracker Green FM, a green fluorescent intracellular probe serving originally for staining mitochondria, seemed especially suitable for labeling NK target cells. Materials and Methods NK target cells were labeled with MitoTracker Green FM. After incubation with effector spleen cells, cell suspensions were stained with propidium iodide (PI), and flow cytometric analysis was performed. RESULTS: MitoTracker Green FM stained efficiently each cell type we assayed, including resting cells, and it was not released from dead cells. NK assays were set up using mouse spleen effector cells and K562 NK target cells. MitoTracker Green FM and PI double staining allowed a discrimination of live and dead target cells, and the cytotoxicity values were in the expected range. Then the method was applied to a less well-known chicken model. We found that chicken-skin fibroblasts had a definite sensitivity to autologous splenic NK cells, sometimes as high as the sensitivity of classic NK targets. CONCLUSIONS: Convenient flow-cytometric NK tests can be performed by MitoTracker Green FM and PI staining. Using this method, we demonstrated that chicken fibroblasts are sensitive to the cytotoxic effect of autologous NK cells. PMID- 11891721 TI - Magnetic field design for selecting and aligning immunomagnetic labeled cells. AB - BACKGROUND: Recently we introduced the CellTracks cell analysis system, in which samples are prepared based on a combination of immunomagnetic selection, separation, and alignment of cells along ferromagnetic lines. Here we describe the underlying magnetic principles and considerations made in the magnetic field design to achieve the best possible cell selection and alignment of magnetically labeled cells. Materials and Methods Computer simulations, in combination with experimental data, were used to optimize the design of the magnets and Ni lines to obtain the optimal magnetic configuration. RESULTS: A homogeneous cell distribution on the upper surface of the sample chamber was obtained with a magnet where the pole faces were tilted towards each other. The spatial distribution of magnetically aligned objects in between the Ni lines was dependent on the ratio of the diameter of the aligned object and the line spacing, which was tested with magnetically and fluorescently labeled 6 microm polystyrene beads. The best result was obtained when the line spacing was equal to or smaller than the diameter of the aligned object. CONCLUSIONS: The magnetic gradient of the designed permanent magnet extracts magnetically labeled cells from any cell suspension to a desired plane, providing a homogeneous cell distribution. In addition, it magnetizes ferro-magnetic Ni lines in this plane whose additional local gradient adds to the gradient of the permanent magnet. The resultant gradient aligns the magnetically labeled cells first brought to this plane. This combination makes it possible, in a single step, to extract and align cells on a surface from any cell suspension. PMID- 11891722 TI - Cell analysis system based on compact disk technology. AB - BACKGROUND: A cell analysis system was developed to enumerate and differentiate magnetically aligned cells selected from whole blood. The cellular information extracted is similar to the readout of musical information from a compact disk (CD). Here we describe the optical design and data processing of the system. The performance of the system is demonstrated using fluorescent-labeled cells and beads. Materials and Methods System performance was demonstrated with 6-microm polystyrene beads labeled with magnetic nanoparticles and allophycocyanin (APC) and immunomagnetically aligned leukocytes, fluorescently labeled with Oxazine750 and CD4-APC, CD8-Cy5.5, and CD14-APC/Cy7 in whole blood. RESULTS: The sensitivity of the system was demonstrated using APC-labeled beads. With this system, beads containing 333 APC molecules could easily be resolved from the background. This level of sensitivity was not achievable with a commercial flow cytometer. A maximum of 20,000 immunomagnetically labeled cells could be aligned and analyzed in between 0.6 m of Ni lines, distributed over a surface area of 18 mm(2) and extracted from a blood volume that depended on the height of the chamber. The utility of the system was demonstrated by performing a three-color CD4-CD8-CD14 assay. CONCLUSIONS: We built a cell analysis system based on immunomagnetic cell selection and alignment and analysis of fluorescent signals employing CD technology that is as good or better than current commercial analyzers. The cell analysis can be performed in whole blood or any other type of cell suspension without extensive sample preparation. PMID- 11891723 TI - Mixing small volumes for continuous high-throughput flow cytometry: performance of a mixing Y and peristaltic sample delivery. AB - BACKGROUND: Online mixing for continuous high-throughput flow cytometry has not been previously described. A simple, general high-throughput method for mixing and delivery of submicroliter volumes in laminar flow at low Reynolds numbers would be widely useful. MATERIALS AND METHODS: We describe a micromixing approach that is compatible with commercial autosamplers, flow cytometry, and other detection schemes that require mixing of components that have been introduced into laminar flow. The scheme is based on a previous approach to high-throughput flow cytometry (HyperCyt, Kuckuck et al.: Cytometry 44:83-90, 2001). We showed that samples from multiwell plates that have been picked up by an autosampler can be separated during delivery by the small air bubbles introduced during the transit of the autosampler probe from well to well. Here, a particle sample flowing continuously is brought together in a Y with reagent samples from wells, which have been separated by bubbles. RESULTS: In the effluent stream, the particles and reagents are mixed, most likely as a result of peristaltic action, and reagents from individual wells can be resolved. The sample volumes that can be mixed with this technology are submicroliter in volume, and samples can be mixed at rates up to at least 100/samples per minute. With the current device, carryover between samples can be eliminated if the mixing system is flushed with several volumes of buffer. The anticipated throughput for screening is expected to be at least 20 samples per minute. CONCLUSIONS: The high-throughput approach and peristaltic mixing in HyperCytTM serve to integrate autosamplers with submicroliter detection volumes for analysis in flow cytometry or in microfluidic channels. PMID- 11891724 TI - Interferometry in flow to sort unstained X- and Y-chromosome-bearing bull spermatozoa. AB - BACKGROUND: It was found earlier that the difference in volume between unstained X- and Y-chromosome-bearing sperm heads could be detected using interference microscopy in visible light. This could be the basis for an alternative to the conventional method to sort X and Y sperm, which uses DNA staining and ultraviolet (UV)-excitation that may be harmful to sperm cells. A novel technique is introduced combining interferometry with flow cytometry. MATERIALS AND METHODS: Interference optics were built into an existing flow cytometer/cell sorter and used to sort fresh unstained bull sperm cells on the basis of their head volume. Sorted fractions were stained with a DNA stain, and reanalyzed using the conventional method. RESULTS: Purities between 60-66% were found in both X- and Y-enriched fractions. It was possible to sort up to 300 cells per second. The system was found to be less sensitive to the orientation of sperm cells than the conventional method. CONCLUSIONS: Interferometry can be combined with flow cytometry and used to obtain significantly enriched fractions of X- and Y-bearing sperm without staining and UV light. Sorting speeds and purities at this point, however, are much lower than with the conventional method. PMID- 11891725 TI - Dye exclusion artifact in flow cytometers. AB - BACKGROUND: Cells exclude their own volume of dye solution in the sample flow which carries them through the flow chamber of the flow cytometer, thereby affecting the otherwise constant signal arising from the fluorescence of this solution. Under certain conditions, this phenomenon may significantly influence the fluorescence signal of the cells. MATERIALS AND METHODS: Using the slit scan technique, we studied this phenomenon as observed for monodisperse polystyrene particles in fluorescein solution. RESULTS: The measurements show that dye solution accumulates just in front of the particle and just behind it, with a relative void in between. This phenomenon is most likely caused by the rapid constriction of the flow as it enters the orifice of the nozzle or flow chamber, giving rise to a pulse of fluorescence which adds to that of the particle or cell itself. The magnitude of this artifact depends on the design and dimensions of the nozzle/flow chamber as well as on the rate of sample flow. CONCLUSIONS: The dye exclusion artifact may affect measurements of cells when they are in a dye solution having a fluorescence per unit volume which is significant compared to that of the cells, especially at low sample flow rates. PMID- 11891727 TI - 2-[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a (HPPH) in a nude rat glioma model: implications for photodynamic therapy. AB - BACKGROUND AND OBJECTIVE: In this study, we evaluated 2-[1-hexyloxyethyl]-2 devinyl pyropheophorbide-alpha (HPPH or Photochlor) as a photosensitizer for the treatment of malignant gliomas by photodynamic therapy (PDT). STUDY DESIGN/MATERIALS AND METHODS: We performed in vivo reflection spectroscopy in athymic rats to measure the attenuation of light in normal brain tissue. We also studied HPPH pharmacokinetics and PDT effects in nude rats with brain tumors derived from stereotactically implanted U87 human glioma cells. Rats implanted with tumors were sacrificed at designated time points to determine the pharmacokinetics of HPPH in serum, tumor, normal brain, and brain adjacent to tumor (BAT). HPPH concentrations in normal brain, BAT and tumor were determined using fluorescence spectroscopy. Twenty-four hours after intravenous injection of HPPH, we administered interstitial PDT treatment at a wavelength of 665 nm. Light was given in doses of 3.5, 7.5 or 15 J/cm at the tumor site and at a rate of 50 mW/cm. RESULTS: In vivo spectroscopy of normal brain tissue showed that the attenuation depth of 665 nm light is approximately 30% greater than that of 630 nm light used to activate Photofrin, which is currently being evaluated for PDT as an adjuvant to surgery for malignant gliomas. The t1/2 of disappearance of drug from serum and tumor was 25 and 30 hours, respectively. CONCLUSION: Twenty four hours after injection of 0.5 mg/kg HPPH, tumor-to-brain drug ratios ranged from 5:1 to 15:1. Enhanced survival was observed in each of the HPPH/PDT-treated animal groups. These data suggest that HPPH may be a useful adjuvant for the treatment of malignant gliomas. PMID- 11891728 TI - Development of a novel indwelling balloon applicator for optimizing light delivery in photodynamic therapy. AB - BACKGROUND AND OBJECTIVE: A human glioma spheroid model is used to investigate the efficacy of different light delivery schemes in 5-aminolevulinic acid (ALA)- mediated photodynamic therapy (PDT). The results provide the rationale for the development of an indwelling balloon applicator for optimizing light delivery. STUDY DESIGN/MATERIALS AND METHODS: Human glioma spheroids were incubated in ALA (100 or 1000 microg /ml-1) for 4 hours and subjected to various light irradiation schemes. In one set of experiments, spheroid survival was monitored as a function of light fluence rate (5-200 mW cm-2). In all cases, spheroids were exposed to fluences of either 25 or 50 J cm-2. In a second study, the effects of repeated weekly PDT treatments, using sub-threshold fluences, were investigated. One group of spheroids was subjected to three treatments using fluences of 12, 12, and 25 J cm-2. Results were compared to spheroids receiving single treatments of either 12 or 25 J cm-2. A fluence rate of 25 mW cm-2 was used for all three groups of spheroids. In all cases, the effect of a given irradiation scheme was evaluated by monitoring spheroid growth. RESULTS: Low fluence rates produce greater cell kill than high fluence rates. The minimum effective fluence rate in human glioma spheroids is approximately 10 mW cm-2. Repeated weekly PDT treatments with sub threshold fluences result in significant cell kill. In spheroids surviving the PDT treatments, growth is suppressed for the duration of the treatment period. CONCLUSION: The results of the in vitro studies support the development of an indwelling balloon applicator for the delivery of light doses in long term multi fractionated PDT regimens. PMID- 11891729 TI - Extended theory of selective photothermolysis: a new recipe for hair cooking? PMID- 11891730 TI - Extended theory of selective photothermolysis. AB - BACKGROUND AND OBJECTIVE: We present a new theory of selective thermal damage of non-uniformly pigmented structures in biological tissues. Spatial separation of the heavily pigmented areas and the target requires limitation of the pigment temperature and heat diffusion from the pigmented to the targeted areas. STUDY DESIGNS/MATERIALS AND METHODS: A concept of selective target damage by heat diffusion is presented for three target geometries: planar, cylindrical, and spherical. An in vitro experiment is described in which the dependence of thermal damage on pulsewidth at constant fluence was evaluated. RESULTS: The in vitro experiment showed that the size of the damage zone for similar hair follicles was pulsewidth-independent over a very broad range of pulsewidths (30-400 ms). We formulated a new theory (extended theory of photothermolysis) to interpret the experimental results. CONCLUSIONS: Based on this new theory, the treatment pulsewidth for non-uniformly pigmented targets is significantly longer than the target thermal relaxation time (TRT). The theory provides new recommendations for photoepilation and photosclerotherapy parameters. PMID- 11891731 TI - Helium-Neon laser irradiation of hepatocytes can trigger increase of the mitochondrial membrane potential and can stimulate c-fos expression in a Ca2+ dependent manner. AB - BACKGROUND AND OBJECTIVE: To gain some insight into the photostimulation of isolated hepatocytes irradiated with Helium-Neon (He-Ne) laser light certain biochemical events were studied with respect to two mechanisms: i) the direct light dependent activation of certain biochemical events investigated in intact cells and isolated mitochondria, ii) the indirect stimulation of processes per se light independent. STUDY DESIGNS/MATERIALS AND METHODS: Irradiation of either isolated hepatocytes or isolated rat liver mitochondria was carried out with He Ne laser (wavelength, 632.8 nm; fluence, 0.24 J cm-2; fluence rate, 12 mW cm-2). Changes in mitochondrial membrane potential in isolated hepatocytes were monitored using the cationic probe safranine. The c-fos expression was studied by Northern blot and immunoblot analysis. RESULTS: As a result of irradiation, increase of the mitochondrial membrane potential was found to occur in irradiated hepatocytes both in the presence or in the absence of CaCl2. The hyperpolarization of the mitochondrial membrane is assumed to cause an increase in mitochondrial Ca2+ uptake that was measured in isolated mitochondria. Finally, an increase in c-fos expression was found in irradiated hepatocytes when incubated in the presence of CaCl2. CONCLUSION: This paper gives additional information on the mechanism by which He-Ne laser light, either directly or in a cascade-like effect dependent on increase in cell Ca2+, can cause cell stimulation. PMID- 11891732 TI - Improvement of macromolecular clearance via lymph flow in hamster gingiva by low power carbon dioxide laser-irradiation. AB - BACKGROUND AND OBJECTIVE: Although therapeutic effects of low-power laser irradiation on periodontal disease have been reported, little is known about the biological effects of laser-irradiation in the gingiva. Recently we reported that topical warming stimulated macromolecular clearance via lymph flow in hamster gingiva. This study was conducted to investigate whether low-power laser irradiation affects macromolecular clearance via the lymph flow in the gingiva. STUDY DESIGN/MATERIALS AND METHODS: We injected 14C-methylated albumin into the mandibular gingiva of anesthetized hamsters followed by topical carbon dioxide (CO2) laser-irradiation (30 seconds, 0.5-1.5 W). We measured the clearance of radiolabeled albumin from the gingiva and its drainage into submandibular lymph nodes during 10 minutes. RESULTS: The clearance of the radiolabeled albumin from the gingiva and the influx into the submandibular lymph nodes were increased by CO2 laser-irradiation. CONCLUSION: Low-power CO2 laser-irradiation improves macromolecular clearance via the lymph flow in hamster gingiva. PMID- 11891733 TI - Laser-induced shock waves enhance sterilization of infected vascular prosthetic grafts. AB - BACKGROUND AND OBJECTIVE: Bacteria that cause infection of vascular prosthetic grafts produce an exopolysaccharide matrix known as biofilm. Growth in biofilms protects the bacteria from leukocytes, antibodies and antimicrobial drugs. Laser generated shock waves (SW) can disrupt biofilms and increase drug penetration. This study investigates the possibility of increasing antibiotic delivery and sterilization of vascular prosthetic graft. STUDY DESIGN/MATERIALS AND METHODS: Strains of Staphylococcus epidermidis and S. aureus were isolated from infected prosthetic grafts obtained directly from patients. Dacron grafts were inoculated with the isolated bacteria, which were allowed to form adherent bacterial colonies. The colonized grafts underwent the following treatments: (a) antibiotic (vancomycin) alone; (b) antibiotic and SW (c) saline only; and (d) saline and SW. Six hours after treatment, the grafts were sonicated, the effluent was cultured and the colony forming units (CFU) were counted. RESULTS: CFU recovered from control grafts colonized by S. epidermidis were comparable: saline, 3.05 x 10(8) and saline+SW 3.31 x 10(8). The number of S. epidermidis CFU diminished to 7.61 x 10(6) after antibiotic treatment but the combined antibiotic+SW treatment synergistically decreased CFU number to 1.27 x 10(4) (P<0.001). S. aureus showed a higher susceptibility to the antibiotic: 2.26 x 10(6) CFU; antibiotic +SW treatment also had an incremental effect: 8.27 x 10(4) CFU (P<0.001). CONCLUSIONS: This study demonstrates that laser-generated shock waves have no effects alone, but can enhance the effectiveness of antibiotics against bacteria associated with prosthetic vascular graft biofilms, suggesting that this treatment may be of value as adjunctive therapy for prosthetic graft infections. PMID- 11891734 TI - Vascular procedures that thermo-coagulate collagen reduce local platelet deposition and thrombus formation: laser and laser-thermal versus balloon angioplasty. AB - BACKGROUND AND OBJECTIVE: Exposure of the arterial wall matrix to blood leads to platelet deposition resulting in thrombosis. Because heat alters tissue matrix we proposed that heating reduces platelet deposition. STUDY DESIGN/MATERIALS AND METHODS: Sixty arterial homografts (15 dogs) were mounted in an arterio-venous "shunt." Interventions included balloon angioplasty (BA), direct laser (LA), laser-thermal (LTA), and combined LTABA. 111Indium-labeled platelets were circulated, radio activity measured, and homografts processed for histology. RESULTS: Radioactivity count (mean+/- SE) at BA sites (13,853+/-3,192 cpm/cm(2)) was greater than LA (7,038+/-981), LTA (5,294 +/-1,145), LTABA (6,176+/-1,571), and control (1,826+/-339), P<0.05. Electron microscopy showed fewer platelets at LA, LTA, and control than BA sites. BA spread the collagen on the arterial lumen while heat gelled collagen and confined it to the arterial media. CONCLUSIONS: Heating the artery and gelling collagen during LA, LTA, or LTABA significantly reduced thrombogenicity. PMID- 11891735 TI - Nanosecond, high-intensity pulsed laser ablation of myocardium tissue at the ultraviolet, visible, and near-infrared wavelengths: in-vitro study. AB - BACKGROUND AND OBJECTIVE: A large number of clinical trials of transmyocardial laser revascularization (TMLR) have been conducted to treat severe ischemic heart diseases. A variety of laser sources have been used or tested for this treatment, however, no comprehensive study has been performed to reveal the mechanism and the optimum laser irradiation condition for the myocardium tissue ablation. There have been reported limited experimental data of the high-intensity pulsed laser ablation of myocardium tissues. STUDY DESIGN/MATERIALS AND METHODS: A 1064-nm Q switched Nd:YAG laser and its 2nd (532 nm), 3rd (355 nm), and 4th (266 nm) harmonics were used for ablation experiments. At each wavelength, 25 laser pulses irradiated the porcine myocardium tissue samples at a constant laser intensity (peak laser power divided by laser spot area) of approximately 2 GW/cm(2) and the ablation depths were measured. During ablation, laser-induced optical and acoustic emissions were measured to investigate the ablation mechanism at each laser wavelength. For the ablated tissues, histological observation was made with a polarization optical microscope. RESULTS: It was shown that the ablation efficiency did not directly depend on the linear absorption coefficient of the tissue; the ablation depth was maximized at 355 and 1064 nm, and minimized at 532 nm. Strong laser-induced optical and acoustic emissions were observed for the 266 and 1064-nm laser irradiations. The histology showed that thermal denaturation of the tissue near the ablation walls decreased with decreasing wavelength for 266, 355, and 532 nm, but it was limited for 1064 nm. CONCLUSION: At the laser intensity of approximately 2 GW/cm(2), ablation characteristics were drastically changed for the different laser wavelengths. The results indicated that for 266, 355, and 532 nm, the tissue removal was achieved mainly through a photothermal process, but for 266 nm the intense laser-induced plasma formation would result in a reduced laser energy coupling to the tissue. For 1064 nm, a photodisruption was most probable as a dominant tissue removal process. Because of the high ablation rate and limited thermal denaturation, the 355- and 1064-nm lasers could be potential laser sources for TMLR, although further investigation is needed to discuss the clinical issues. PMID- 11891736 TI - Unique features of optical scanning, single fiber endoscopy. AB - BACKGROUND AND OBJECTIVE: To advance the field of minimally invasive medical procedures, an ideal endoscope should provide high-resolution images with variable magnification from an ultra-thin package, while adding depth cues and integrating optical diagnoses and therapies. Satisfying all these requirements is extremely difficult using commercial endoscopes. A new imaging technology is introduced that uses directed laser illumination, which is scanned at the distal end of a flexible endoscope. STUDY DESIGN/MATERIALS AND METHODS: A single-mode optical fiber is driven in vibratory resonance using a piezoelectric actuator. The emitted laser light is scanned in two-dimensions over test specimens. Digital images are constructed by detecting optical power one pixel at a time. RESULTS: Unique features of the fiber scanning scope are rapidly changing magnification, enhanced topographic detail, and concurrent fluorescence imaging, which are demonstrated and discussed. CONCLUSION: This fiber scanning scope has the potential for pixel-accurate delivery of high quality laser radiation, allowing the future integration of imaging with diagnosis and therapy. PMID- 11891737 TI - Diode-pumped fiber lasers: a new clinical tool? AB - BACKGROUND AND OBJECTIVE: Diode-pumped fiber lasers are a compact and an efficient source of high power laser radiation. These laser systems have found wide recognition in the area of lasers as a result of these very practical characteristics and are now becoming important tools for a large number of applications. In this review, we outline the basic physics of fiber lasers and illustrate how a number of clinical procedures would benefit from their employment. STUDY DESIGN/MATERIALS AND METHODS: The pump mechanisms, the relevant pump and laser transitions between the energy levels, and the main properties of the output from fiber lasers will be briefly reviewed. The main types of high power fiber lasers that have been demonstrated will be examined along with some recent medical applications that have used these lasers. We will also provide a general review of some important medical specialties, highlighting why these fields would gain from the introduction of the fiber laser. RESULTS/CONCLUSION: It is established that while the fiber laser is still a new form of laser device and hence not commercially available in a wide sense, a number of important medical procedures will benefit from its general introduction into medicine. With the number of medical and surgical applications requiring high power laser radiation steadily increasing, the demand for more efficient and compact laser systems providing this capacity will grow commensurately. The high power fiber laser is one system that looks like a promising modality to meet this need. PMID- 11891738 TI - Optimal methods for fluorescence and diffuse reflectance measurements of tissue biopsy samples. AB - BACKGROUND AND OBJECTIVE: In developing fluorescence spectroscopy systems for the in vivo detection of pre-cancer and cancer, it is often necessary to perform preliminary testing on tissue biopsies. Current standard protocols call for the tissue to be immediately frozen after biopsy and later thawed for spectroscopic analysis, but this process can have profound effects on the spectroscopic properties of tissue. This study investigates the optimal tissue handling methods for in vitro fluorescence spectroscopy studies. STUDY DESIGN/MATERIALS AND METHODS: The epithelial tissue of the Golden Syrian hamster cheek pouch was used in this study. Three specific experiments were carried out. First, the fluorescence properties of tissues in vivo and of frozen and thawed tissue biopsies were characterized at multiple excitation wavelengths spanning the ultraviolet-visible (UV-VIS) spectrum. Next, comparison of tissue fluorescence emission spectra in vivo, ex vivo (immediately after biopsy), and after the freeze and thaw process were systematically carried out at the excitation wavelengths corresponding to the previously identified fluorescence peaks. Lastly, intensities at the excitation and emission wavelength pairs corresponding to the fluorescence peaks were measured as a function of time after biopsy. Diffuse reflectance measurements over the UV-VIS spectrum were also made to evaluate the effects of oxygenation, blood volume, and scattering on the tissue fluorescence at these different excitation-emission wavelengths. RESULTS: This study indicates that the freezing and thawing process produces a significant deviation in intensity and lineshape relative to the in vivo fluorescence emission spectral data over the entire UV-VIS range between 300 and 700 nm. By contrast, examination of ex vivo emission spectra reveals that it closely preserves both the intensity and lineshape of the in vivo emission spectra except between 500 and 700 nm. The observed deviations can be explained by the diffuse reflectance measurements, which suggest increased hemoglobin deoxygenation and wavelength dependent changes in scattering in ex vivo tissues, and increased total hemoglobin absorption in the frozen and thawed samples. Furthermore, it was found that over a time window of 1.5 hours, spectroscopic changes brought about by degradation of the tissue due to biopsy or other factors are significantly smaller (10-30% variations in intensity) than those associated with the freezing and thawing process (50-70% decrease in intensity). CONCLUSIONS: It was found that the effects of freezing and thawing on the fluorescence properties of tissue are greater than any changes brought about by degradation of tissue over a time frame of 90 minutes after biopsy. Performing ex vivo fluorescence measurements within a reasonable time window has the advantage of more accurately reproducing the clinically relevant in vivo conditions in the case of the hamster cheek pouch tissue. Therefore, in tissue biopsy studies, the tissue sample should ideally be maintained in an unfrozen state prior to measurement. PMID- 11891739 TI - Propylene glycol as a contrasting agent for optical coherence tomography to image gastrointestinal tissues. AB - BACKGROUND AND OBJECTIVE: Optical coherence tomography (OCT) is a recently developed imaging technique that has the potential to advance the early diagnosis of diseases in the human gastrointestinal (GI) tract. How ever, the high scattering nature of GI tissue limits its imaging depth and contrast. For more effective diagnosis using OCT, a concurrent improvement of imaging depth and contrast is, therefore, needed. In this work, we investigate the administration of chemical agents to the tissue as a means of improving the capability of OCT imaging of clinically relevant microstructures of the GI tract. STUDY DESIGN/MATERIALS AND METHODS: Normal human GI tissues, including stomach and oesophagus were obtained from patients in hospital, and were imaged with OCT within 0.5-2 hours of removal. Immediately after the first imaging of the specimens with OCT, about 0.5 ml of 80% propylene glycol solution was applied onto the tissue surface and the tissue allowed to absorb the chemical compounds for 20 minutes. Another image was then taken at the same position. The specimens were then embedded and stained in preparation for histologic evaluation. Co registration of the images obtained using OCT before and after the topical application of the propylene glycol solution, and standard histopathologic processing provided basis for comparison. RESULTS: More detailed micro structures, including the basal layer position and the cellular composition of the mucosal layer of GI tract tissues were observed after the topical application of propylene glycol solution, while these structures were not resolvable in the conventional OCT images. CONCLUSIONS: Propylene glycol could be used as a contrasting agent for OCT imaging of human GI tract tissues, allowing an increased capability of OCT for rapid clinical diagnosis in vivo. PMID- 11891740 TI - Transscleral optical coherence tomography--an experimental study in ex-vivo human eyes. AB - BACKGROUND AND OBJECTIVE: To evaluate the potentials of a 1310-nm optical coherence tomography (OCT) system to penetrate the highly backscattering sclera in enucleated human eyes and provide visualization of intraocular structures by transscleral imaging. STUDY DESIGN/MATERIALS AND METHODS: OCT-images were generated by an experimental prototype (Medical Laser Center, Lubeck, Germany) using a superluminescence diode with a wavelength of 1310 nm. OCT-images were taken from two enucleated human eyes using 100-200 axial scans with 60 Hz line scan frequency and compared to subsequent histologic sections. RESULTS: Transscleral OCT allowed penetration of the sclera and the anterior chamber angle could be completely identified. Some change within the anterior eye segment could be demonstrated with high accuracy. Additionally, limited demonstration of the ciliary body region was achieved. Due to limited signal intensity no detailed imaging of the pars plana and pars plicata region was possible. However, more posterior measurements allowed transscleral visualization of a retinal detachment. CONCLUSIONS: OCT using lightsources with a wavelength longer than that used in conventional OCT provides a promising imaging technique at high resolution allowing transscleral imaging of the anterior eye segment. PMID- 11891741 TI - Ultrashort pulse laser ossicular ablation and stapedotomy in cadaveric bone. AB - BACKGROUND AND OBJECTIVE: The purpose of this study was to evaluate the ablation of ossicular tissue using a 1,053 nm Ti:Sapphire chirped pulse amplifier laser system configured to deliver ultrashort pulses of 350 femtoseconds (fs) (3.5x10( 13) seconds) in cadaver temporal bone. STUDY DESIGN/MATERIALS AND METHODS: Ablation of the formalin-fixed incus and stapes was performed using an ultrashort pulse laser (USPL) (0.4 mm beam diameter, pulse fluence of 2.0 J/cm2, and pulse repetition rate of 10 Hz). The ablation rate was measured using optical micrometry, and crater surface morphology examined using scanning electron microscopy. RESULTS: The laser produced precise bone ablation at a rate of 1.26 microm/pulse, with almost no evidence of thermal damage, and very little evidence of photomechanical injury. CONCLUSIONS: Ultrashort pulse lasers may provide a useful clinical tool for otologic and skull base surgery, where precise hard tissue ablation is required adjacent to critical structures. PMID- 11891742 TI - Electrophysiologic effect of gallium arsenide laser on frog gastrocnemius muscle. AB - BACKGROUND AND OBJECTIVE: In this study, the effect of low energy Gallium arsenide (GaAs) laser irradiation on the compound action potential of frog gastrocnemius muscle were investigated. STUDY DESIGN/MATERIALS AND METHODS: Sixty frogs were divided into different six dose groups: laser 1 (1 Hz), laser 2 ( 4 Hz), laser 3 (16 Hz), laser 4 (64 Hz), laser 5 (128 Hz), and laser 6 (1,000 Hz, DC, continue) (in each group n=10). Low energy GaAs laser (wavelenght: 904 nm, pulsed duration: 220 nanoseconds, peak power per pulse: 27 W, total applied energy density: 0.001-25.7 J/cm2) was used for the experiment. Compound muscle action potentials were recorded before laser irradiation and these data were accepted as control group. After recording the control data, each muscle was irradiated by the laser. Action potentials were recorded at 1, 5, 10, 15, and 20 minutes of irradiation time in each group by using standartized needle electromyography and nerve conduction study techniques. Distal motor latency, peak to peak amplitude, area, and total duration of action potential were measured. Repeated measures analysis of variance were used for the statistical evaluation. RESULTS: No significant differences were detected between control and laser dose groups in muscle action potential parameters. CONCLUSIONS: This study revealed that at the different repetition rate and exposure time, low energy GaAs laser does not have any significant effect on frog gastrocnemius action potential. PMID- 11891743 TI - Effects of laser-induced thermotherapy (LITT) on proliferation and apoptosis of glioma cells in rat brain transplantation tumors. AB - BACKGROUND AND OBJECTIVES: Laser-induced thermotherapy (LITT) is an approach to the treatment of brain tumors especially in poorly accessible regions. Its clinical applicability with tumor cell destruction has been shown in several studies. However, no data are known about specific effects on tumors cells due to LITT in the time course of the lesion. STUDY DESIGN/MATERIALS AND METHODS: LITT was performed in adult Lewis rats with implanted glioma cells in the brain using a standard exposure of 3 W for 30 seconds. Before and following LITT, neoplastic lesions were monitored by MRI. Proliferation of implanted cells and gliosis were assessed by several histological techniques and immunohistochemistry. Apoptosis was detected by TUNEL staining. RESULTS: Our experiments show a destruction of neoplastic cells by LITT but surviving tumor cells at the margin of the lesion. Apoptosis was detected following LITT restricted to residual neoplastic cells. Marginal survival of tumor cells lead to a secondary outgrowth into the necrotic lesion adjacent to sprouting capillaries. CONCLUSIONS: LITT is a suitable technique for the treatment of brain neoplasms. However, further investigations are necessary to prevent tumor recurrences after LITT. PMID- 11891744 TI - Effect of simulated CO2 and GaAlAs laser surface decontamination on temperature changes in Ti-plasma sprayed dental implants. AB - BACKGROUND AND OBJECTIVE: To investigate and compare temperature elevations at the implant-bone interface during simulated implant surface decontamination with a CO2 and a GaAlAs laser. STUDY DESIGN/MATERIALS AND METHODS: Stepped cylinder implants (Frialit 2) Friadent GmbH, Mannheim, Germany) with a Titanium plasma sprayed surface were inserted into bone blocks cut from pig femurs. An artificial periimplant bone defect provided access for laser irradiation in the coronal third. Both lasers were operated at 1.0-2.5 W in the cw-mode. The bone block was placed into a 37 degrees C water bath in order to simulate in vivo thermal conductivity and diffusitivity of heat. K-type thermocouples connected to a digital meter were used to register temperature changes at the periimplant bone. RESULTS: In mean, the critical threshold of 47 degrees C was exceeded after 8 seconds at a power output of 2.5 W, 13 seconds at 2.0 W, 18 seconds at 1.5 W, and 42 seconds at 1.0 W with the GaAlAs laser and 15 seconds (2.5 W), 23 seconds (2.0 W), 35 seconds (1.5 W), and 56 seconds (1.0 W) with the CO2 laser. At equal energy fluence, GaAlAs laser irradiation induced significantly higher temperature elevations than CO2 laser irradiation. CONCLUSIONS: In an energy dependent manner implant surface decontamination with both laser types must be limited in time to allow the implant and bone to cool down. Clinical guidelines are presented to avoid tissue damage. PMID- 11891745 TI - Selective removal of residual composite from dental enamel surfaces using the third harmonic of a Q-switched Nd:YAG laser. AB - BACKGROUND AND OBJECTIVE: Conventional methods of residual composite removal after the debonding of orthodontic brackets involve the use of abrasives that damage the underlying enamel. The objective of this study was to demonstrate that 355-nm laser pulses with a pulse width of 10 ns are well suited for the removal of composite through selective laser ablation. STUDY DESIGN/MATERIALS AND METHODS: The residual composite remaining on the surface of extracted human third molars and bovine incisors was removed using multiple laser pulses from the third harmonic (355-nm) of a Q-switched Nd:YAG laser. RESULTS: There is selective ablation of composite from the enamel surface without any discernable damage to the underlying enamel. CONCLUSION: This study demonstrates that 355-nm, 10 ns laser pulses can be used for the selective ablation of dental composite without thermal or mechanical damage to the underlying enamel. PMID- 11891746 TI - Ultrastructural changes of human dentin after irradiation by Nd:YAG laser. AB - BACKGROUND AND OBJECTIVE: The use of Nd:YAG laser has been proposed for endodontic treatment. However, its ability to reduce dentin permeability, which is important for the success of root canal treatment, remains controversial. STUDY DESIGN/MATERIALS AND METHODS: Nd:YAG laser irradiation was performed in pulsed mode on human dentin. The parameters were: pulse energy (100 mJ), rate (10 pps), and total irradiation time (4 seconds). The crystalline phases, electron diffraction patterns, morphology, and microstructure of specimens after laser irradiation were observed by dark-field emission transmission electron microscope (TEM). RESULTS: Three ultrastructural zones could be delineated in the dentin: (1) an outer zone with an ordered columnar structure composed of hydroxyapatite and beta-tricalcium phosphate, (2) an intermediate zone composed of an amorphous substance (about 40-70 nm in diameter), and (3) an inner zone of well crystallized hydroxyapatite grains. These three zones were free of pores or voids. CONCLUSIONS: Our study demonstrated that laser-irradiation might be used to reduce dentin permeability. PMID- 11891747 TI - Study of high- and low-current-configuration homes from the 1988 Denver Childhood Cancer Study. AB - An epidemiological study conducted by Savitz et al. reported that residential wire codes were more strongly associated with childhood cancer than were measured magnetic fields, a peculiar result because wire codes were originally developed to be a surrogate for residential magnetic fields. The primary purpose of the study reported here, known as the Back to Denver (BTD) study, was to obtain data to help in the interpretation of the original results of Savitz et al. The BTD study included 81 homes that had been occupied by case and control subjects of Savitz et al., stratified by wire code as follows: 18 high current configuration (HCC) case homes; 20 HCC control homes; 20 low current configuration (LCC) case homes; and 23 LCC control homes. Analysis of new data acquired in these homes led to the following previously unpublished conclusions. The home-averaged (i.e., mean of fields measured in subjects' bedrooms, family/living rooms, and rooms where meals normally eaten) spot 60 Hz, 180 Hz, and harmonic (i.e., 60-420 Hz) magnetic fields were associated with wire codes. The 180 Hz and harmonic components, but not the 60 Hz component, were associated with case/control status. Measured static magnetic fields were only weakly correlated (rapproximately 0.2) between rooms in homes. The BTD data provide little support for, but are too sparse to definitively test, the 1995 resonance hypothesis proposed by Bowman et al. Case and control homes had similar concentrations of copper in their tap water. Copper concentration was not associated with wire codes nor with the level of electric current carried by a home's water pipe. These results of the BTD study suggest that future case/control studies investigating power frequency magnetic fields might wish to include measurements of 180 Hz or harmonic magnetic fields in order to examine their associations (if any) with disease status. PMID- 11891748 TI - Brief exposure to a 50 Hz, 100 microT magnetic field: effects on reaction time, accuracy, and recognition memory. AB - The present study investigated both the direct and delayed effects of a 50 Hz, 100 microT magnetic field on human performance. Eighty subjects completed a visual duration discrimination task, half being exposed to the field and the other half sham exposed. The delayed effects of this field were also examined in a recognition memory task that followed immediately upon completion of the discrimination task, Unlike our earlier studies, we were unable to find any effects of the field on reaction time and accuracy in the visual discrimination task. However, the field had a delayed effect on memory, producing a decrement in recognition accuracy. We conclude that after many years of experimentation, finding a set of magnetic field parameters and human performance measures that reliably yield magnetic field effects is proving elusive. Yet the large number of significant findings suggests that further research is warranted. PMID- 11891749 TI - Temporal trends and misclassification in residential 60 Hz magnetic field measurements. AB - This research addressed the question of how well measurement data collected during a single visit, made at an arbitrary hour of day, day of week, and season, estimate longer term residential 60 Hz magnetic field levels. We made repeat spot and 24 h measurements in 51 children's home, located in the Detroit, MI, and the Minneapolis-St. Paul, MN metropolitan areas, on a regular bimonthly schedule over a 1 year period, as well as a single 2 week measurement, for total of eight visits, producing 21 days of data for each residence. We defined the long term estimate (LTE) as the geometric mean of all available 24 h geometric means from the first six bimonthly visits. The LTE served as the reference level for assessing seasonal, day of week, and diurnal effects, as well as the potential for misclassification. We found a small, but statistically significant (P<.05), seasonal effect, with levels approximately 3% lower than the LTE in the spring and about 4% greater during the summer. No effect was found for day of week. However, we did find a systematic and appreciable diurnal effect, suggesting that, for example, an evening spot measurement may overestimate the LTE by 20% or more. We also assessed how well the 24 h measurement from the last visit, which was not used in calculation of the LTE, estimated the LTE. We found a high degree of correlation (r=.92) and fair to good agreement using four exposure categories (kappa=.53). Thus, the 24 h measurement appears to be a satisfactory LTE estimator. However, this finding must be interpreted with caution since considerable unexplained variability was present among the repeat 24 h measurements in about one-third of the homes. While the 2 week measurement does somewhat decrease exposure misclassification, its added intrusiveness and cost are likely to outweigh the improved precision. PMID- 11891750 TI - Carcinogenicity test in B6C3F1 mice after parental and prenatal exposure to 50 Hz magnetic fields. AB - Some epidemiological studies suggest association of childhood cancer with occupational exposure of the parents to magnetic fields. To test this relationship, 50 each of C57BL/6J female and C3H/HeJ male mice were exposed for 2 and 9 weeks, respectively, to 50 Hz sham (group A), 0.5 (group B), and 5 mT (group C) sinusoidal alternating magnetic fields. They were mated under the exposure for up to 2 weeks, and the exposure was continued until parturition. All the B6C3F1 offspring, without adjusting numbers of animals, were clinically observed without exposure to magnetic field for a nominal 78 weeks from 6-8 weeks of age after weaning and then euthanized for pathological examination according to a routine carcinogenicity test. 540 pups entered the test, and the survival rate was 96.7%. No F1 mouse died of tumoral diseases before a male in A group died of stomach cancer at 43 weeks of age. The first animal death in the exposed groups due to tumor occurred at 71 weeks of age. Eighteen animals died before necropsy at 84-86 weeks of age. No significant difference was detected in the final number of survivors and incidence of tumors between groups A and B, or A and C. Concerning reproduction total implants in group B were less than in group A and the difference was on the borderline of significance (P=.05). This difference was not reproduced in a later duplicate experiment. PMID- 11891751 TI - Electrorotation of axolotl embryos. AB - The frequency dependent dielectric properties of individual axolotl embryos (Ambystoma mexicanum) were investigated experimentally utilizing the technique of electrorotation. Individual axolotl embryos, immersed in low conductivity media, were subjected to a known frequency and fixed amplitude rotating AC electric field and the ensuing rotational motion of the embryo was monitored using a conventional optical microscope. None of the embryos in the pregastrulation or neurulation stages of development exhibited any rotational motion over the field frequency range (10 Hz-5 MHz). Over the same frequency range, the embryos in the gastrulation stage of development exhibited both co-field and counterfield rotation over different ranges of the applied field frequency. Typically, the counterfield rotation exhibited a peak in the rotation spectrum at similar 1 KHz while the co-field peak was located at similar 1-2 MHz. The rotational spectral data was analyzed using a multishelled spherical embryo model to determine the electrical character of embryos during the early development stages (Stages 5-16; i.e., 16 cell through open neural plate stages). PMID- 11891752 TI - Effect of 0.25 T static magnetic field on microcirculation in rabbits. AB - We showed previously in rabbits that 0.2 and 0.35 T static magnetic field (SMF) modulated systemic hemodynamics by arterial baroreceptors. We now have measured the effect of 0.25 T SMF on microcirculation within cutaneous tissue of the rabbit ear lobe by the rabbit ear chamber (REC) method. Forty experimental runs (20 controls and 20 SMF) were carried out in eight different rabbits with an equal number of control and SMF experiments on each individual. Rabbits were sedated by pentobarbital sodium (5 mg/kg/h, i.v.) during the entire 80 min experiment. SMF was generated by four neodium-iron-boron alloy (Nd2-Fe14-B) magnets (15 x 25 x 30 mm, Neomax, PIP - Tokyo Co., Ltd., Tokyo, Japan), positioned around the REC on the observing stage of an optical microscope. The direct intravital microscopic observation of the rabbit's ear microvascular net, along with simultaneous blood flow measurement by microphotoelectric plethysmography (MPPG), were performed PRE (20 min, baseline), DURING (40 min), and POST (20 min) magnetic field exposure. The control experiments were performed under the same conditions and according to the same time course, but without magnetic field. Data were analyzed comparing MPPG values and percent change from baseline in the same series, and between corresponding sections of control and SMF runs. In contrast to control series (100+/-0.0%-90.0+/-5.4%-87.7+/-7.1%, PRE EXPOSURE-POST), after magnetic field exposure we observed increased blood flow (100+/-0.0%-117.8+/-9.6%*-113.8+/-14.0%, *P<0.05) which gradually decreased after exposure cessation. We propose that long exposure of a high level nonuniform SMF probably modifies microcirculatory homeostasis through modulation of the local release of endothelial neurohumoral and paracrine factors that act directly on the smooth muscle of the vascular wall, presumably by affecting ion channels or second messenger systems. PMID- 11891753 TI - Increased mouse survival, tumor growth inhibition and decreased immunoreactive p53 after exposure to magnetic fields. AB - The possibility that magnetic fields (MF) cause antitumor activity in vivo has been investigated. Two different experiments have been carried out on nude mice bearing a subcutaneous human colon adenocarcinoma (WiDr). In the first experiment, significant increase in survival time (31%) was obtained in mice exposed daily to 70 min modulated MF (static with a superimposition of 50 Hz) having a time average total intensity of 5.5 mT. In the second independent experiment, when mice bearing tumors were exposed to the same treatment for four consecutive weeks, significant inhibition of tumor growth (40%) was reported, together with a decrement in tumor cell mitotic index and proliferative activity. A significant increase in apoptosis was found in tumors of treated animals, together with a reduction in immunoreactive p53 expression. Gross pathology at necroscopy, hematoclinical/hematological and histological examination did not show any adverse or abnormal effects. Since pharmacological rescue of mutant p53 conformation has been recently demonstrated, the authors suggest that MF exposure may obtain a similar effect by acting on redox chemistry connected to metal ions which control p53 folding and its DNA-binding activity. These findings support further investigation aimed at the potential use of magnetic fields as anti cancer agents. PMID- 11891754 TI - Relative contribution of residential and occupational magnetic field exposure over twenty-four hours among people living close to and far from a power line. AB - This study sought to estimate the relative contribution of exposure to 50 Hz magnetic fields experienced at home, at work/school, or elsewhere to the total exposure over 24 hr. Personal exposure meters were carried by 97 adults and children in the Stockholm area. About half of the subjects lived close (<50 m) to a transmission line and half far (>100 m) away. Spot measurements and calculations for the residential exposure were also made. For subjects living<50 m from the line, the exposure at home contributed about 80% of the total magnetic field exposure, measured in mT-hours. Adults living far away experienced only 38% of the total exposure at home, but children still received 55%. Subjects with low time-weighted average (TWA) exposure both at home and at work spent 84% of their time in fields <0.1 microT, and those with high TWA at both locations spent 69% of their time in fields > or = 0.2 microT. This contrast was diluted if only exposure at one location was considered. For spot measurements and calculations of the residential exposure, both sensitivity and specificity was good. However, the intermediate field exposure category (0.1-0.19 microT) showed poor correlation to the 24 hr personal measurements. PMID- 11891755 TI - One week of exposure to 50 Hz, vertical magnetic field does not reduce urinary 6 sulphatoxymelatonin excretion of male wistar rats. AB - The effect of exposure to 100 or 50 microT, 50 Hz, vertical magnetic field on the excretion of 6-sulphatoxymelatonin (6SM) in the nocturnal urine of rats was studied. Twelve male Wistar rats were kept under 12:12 hr light:dark conditions. The nocturnal urine of animals was collected in metabolic cages over 4 consecutive weeks. The concentration of 6SM in the rat urine was measured by 125I radioimmunoassay and normalized to creatinine concentration. After the first week of urine collection, 6 rats were exposed to 100 microT or 50 microT flux density magnetic fields (MF) for 8 hr daily for 1 week. It was found that the excretion of the primary metabolite of melatonin in the urine, 6SM, did not show statistically significant changes during and after magnetic field exposure. PMID- 11891756 TI - The tree of life and the rock of ages: are we getting better at estimating phylogeny? AB - In a recent paper,(1) palaeontologist Mike Benton claimed that our ability to reconstruct accurately the tree of Life may not have improved significantly over the last 100 years. This implies that the cladistic and molecular revolutions may have promulgated as much bad "black box" science as rigorous investigation. Benton's assessment was based on the extent to which cladograms (typically constructed with reference only to distributions of character states) convey the same narrative as the geochronological ages of fossil taxa (an independent data set). Fossil record quality varies greatly between major clades, and the palaeontological dating "yardstick" may be more appropriate for some groups than others. PMID- 11891757 TI - Vetulicolians--are they deuterostomes? chordates? AB - A recent paper by Shu et al.(1) reinterprets the fossil Vetulicola and related forms, all from the Lower Cambrian, as basal deuterostomes, assigning them their own phylum, Vetulicolia. Their conclusion is based on the presence of structures resembling gill slits and a trunk-like region that shows evidence of segmentation. This report summarizes the fossil evidence for their interpretation and evaluates a possible alternative, that vetulicolians may instead be tunicate like chordates. Implications for our understanding of the nature of the primitive deuterostome (and chordate) body plan are discussed. . PMID- 11891758 TI - Synaptic modification in neural circuits: a timely action. AB - Long-term modification of synaptic strength is thought to be the basic mechanism underlying the activity-dependent refinement of neural circuits and the formation of memories engrammed on them. Studies ranging from cell culture preparations to humans subjects indicate that the decision of whether a synapse will undergo strengthening or weakening critically depends on the temporal order of presynaptic and postsynaptic activity. At many synapses, potentiation will be induced only when the presynaptic neuron fires an action potential within milliseconds before the postsynaptic neuron fires, whereas weakening will occur when it is the postsynaptic neuron that fires first. Such processes might be important for the remodeling of neural circuits by activity during development and for network functions such as sequence learning and prediction. Ultimately, this synaptic property might also be fundamental for the cognitive process by which we structure our experience through cause and effect relations. PMID- 11891759 TI - A fresh look at the role of CaMKII in hippocampal synaptic plasticity and memory. AB - Advances in molecular, genetic, and cell biological techniques have allowed neuroscientists to delve into the cellular machinery of learning and memory. The calcium and calmodulin-dependent kinase type II (CaMKII) is one of the best candidates for being a molecular component of the learning and memory machinery in the mammalian brain. It is present in abundance at synapses and its enzymatic properties and responsiveness to intracellular Ca(2+) fit a model whereby Ca(2+) currents activate the kinase and lead to changes in synaptic efficacy. Indeed, such plastic properties of synapses are thought to be important for memory formation. Genetic analysis of the alpha isoform of CaMKII in mice support the hypothesis that CaMKII signaling is required to initiate the formation of new spatial memories in the hippocampus. CaMKII is also required for the correct induction of long-term potentiation (LTP) in the hippocampus, consistent with the widely held belief that LTP is a mechanism for learning and memory. Recent cell biological, genetic, and physiological analyses suggest that one of the cellular explanations for LTP and CaMKII function might be the trafficking of AMPA-type receptors to synapses in response to neural activity. PMID- 11891760 TI - Plant chromatin: development and gene control. AB - It is increasingly clear that chromatin is not just a device for packing DNA within the nucleus but also a dynamic material that changes as cellular environments alter. The precise control of chromatin modification in response to developmental and environmental cues determines the correct spatial and temporal expression of genes. Here, we review exciting discoveries that reveal chromatin participation in many facets of plant development. These include: chromatin modification from embryonic and meristematic development to flowering and seed formation, the involvement of DNA methylation and chromatin in controlling invasive DNA and in maintenance of epigenetic states, and the function of chromatin modifying and remodeling complexes such as SWI/SNF and histone acetylases and deacetylases in gene control. Given the role chromatin structure plays in every facet of plant development, chromatin research will undoubtedly be integral in both basic and applied plant biology. PMID- 11891761 TI - Diversity in the mechanisms of gene regulation by estrogen receptors. AB - The sequencing of the human genome has opened the way for using bioinformatics to identify sets of genes controlled by specific regulatory signals. Here, we review the unexpected diversity of DNA response elements mediating transcriptional regulation by estrogen receptors (ERs), which control the broad physiological effects of estrogens. Consensus palindromic estrogen response elements are found in only a few known estrogen target genes, whereas most responsive genes contain only low-affinity half palindromes, which may also control regulation by other nuclear receptors. ERs can also regulate gene expression in the absence of direct interaction with DNA, via protein-protein interactions with other transcription factors or by modulating the activity of upstream signaling components, thereby significantly expanding the repertoire of estrogen-responsive genes. These diverse mechanisms of action must be taken into account in screening for potential estrogen-responsive sequences in the genome or in regulatory regions of target genes identified by expression profiling. PMID- 11891762 TI - O6-alkylguanine-DNA alkyltransferase: role in carcinogenesis and chemotherapy. AB - The DNA in human cells is continuously undergoing damage as consequences of both endogenous processes and exposure to exogenous agents. The resulting structural changes can be repaired by a number of systems that function to preserve genome integrity. Most pathways are multicomponent, involving incision in the damaged DNA strand and resynthesis using the undamaged strand as a template. In contrast, O(6)-alkylguanine-DNA alkyltransferase is able to act as a single protein that reverses specific types of alkylation damage simply by removing the offending alkyl group, which becomes covalently attached to the protein and inactivates it. The types of damage that ATase repairs are potentially toxic, mutagenic, recombinogenic and clastogenic. They are generated by certain classes of carcinogenic and chemotherapeutic alkylating agents. There is consequently a great deal of interest in this repair system in relation to both carcinogenesis and cancer chemotherapy. PMID- 11891763 TI - Wrestling with pleiotropy: genomic and topological analysis of the yeast gene expression network. AB - The vast majority (>95%) of single-gene mutations in yeast affect not only the expression of the mutant gene, but also the expression of many other genes. These data suggest the presence of a previously uncharacterized "gene expression network"--a set of interactions between genes which dictate gene expression in the native cell environment. Here, we quantitatively analyze the gene expression network revealed by microarray expression data from 273 different yeast gene deletion mutants.(1) We find that gene expression interactions form a robust, error-tolerant "scale-free" network, similar to metabolic pathways(2) and artificial networks such as power grids and the internet.(3-5) Because the connectivity between genes in the gene expression network is unevenly distributed, a scale-free organization helps make organisms resistant to the deleterious effects of mutation, and is thus highly adaptive. The existence of a gene expression network poses practical considerations for the study of gene function, since most mutant phenotypes are the result of changes in the expression of many genes. Using principles of scale-free network topology, we propose that fragmenting the gene expression network via "genome-engineering" may be a viable and practical approach to isolating gene function. PMID- 11891764 TI - Mitochondrial mutations may drive Y chromosome evolution. AB - The human Y chromosome contains very low levels of nucleotide variation. It has been variously hypothesized that this invariance reflects historic reductions in the human male population, a very recent common ancestry, a slow rate of molecular evolution, an inability to evolve adaptively, or frequent selective sweeps acting on genes borne on the Y chromosome. We propose an alternative theory in which human Y chromosome evolution is driven by mutations in the maternally inherited mitochondrial genome, which impair male fertility and ultimately lead to a reduction in the effective population size (N(e)) and consequently the variability of the Y chromosome. PMID- 11891765 TI - Vascular endothelial growth factor (VEGF), a survival factor for tumour cells: implications for anti-angiogenic therapy. AB - Angiogenesis is central to both the growth and metastasis of solid tumours. Anti angiogenic strategies result in blood vessel regression accompanied by tumour cell apoptosis. Radiotherapy and many chemotherapeutic agents kill tumours by inducing apoptotic cell death. We propose that, in addition to its role as an angiogenic factor, vascular endothelial growth factor (VEGF) can act as a survival factor for tumour cells protecting them from apoptosis. Thus anti angiogenics, in particular those directed against VEGF, have multiple anti-tumour effects. We suggest that anti-VEGF strategies prevent vessel growth and block a tumour cell survival factor, VEGF, rendering tumour cells more sensitive to chemotherapy and radiotherapy. In addition, as chemotherapy and radiotherapy have been shown to increase VEGF expression, anti-VEGF strategies may overcome therapy induced tumour cell resistance. PMID- 11891766 TI - Mechanisms of the antitumoral effect of lipid A. AB - Bacterial lipopolysaccharide (LPS) and its active component, lipid A, have been used either alone or as adjuvant in therapeutic anticancer vaccines. Lipid A induces various transcription factors via intracellular signaling cascades initiated by their receptor CD14-TLR4. These events lead to the synthesis of cytokines, which either have direct cytotoxic effect or stimulate the immune system. Their antitumoral effect has been demonstrated in animal models as well as clinical trials. Studies in animal models showed that their antitumoral effect relies mostly on the generation of an effective immune response. In humans, the antitumoral effect was correlated with an antibody response and cell-mediated cytotoxicity. So far, some encouraging results have been achieved in phase I and II clinical trials with regards to response and stabilization of the disease, but an expansion of the studies and trials is needed to find the best conditions for their clinical application. PMID- 11891767 TI - Creating bridges or rifts? Developmental systems theory and evolutionary developmental biology. PMID- 11891768 TI - Nogos and the Nogo-66 receptor: factors inhibiting CNS neuron regeneration. AB - The recently cloned gene Nogo, whose alternative splice products correspond to the antigenic target of the central nervous system (CNS) regeneration enhancing monoclonal antibody IN-1, codes for membrane proteins enriched in brain, particularly in oligodendrocytes. The 66-amino acid extracellular domain of Nogo (Nogo-66) interacts with a high-affinity receptor (NgR), a glycosylphosphatidylinositol (GPI)-linked protein with multiple leucine-rich repeats. The amino terminal cytoplasmic domain of Nogo appears to have a general cellular growth inhibitory effect. Nogo-66, on the other hand, specifically retards neurite outgrowth and induces growth cone collapse, possibly through its interaction with NgR and as yet unidentified transmembrane coreceptors. Recent results also suggest that Nogo expression may induce apoptosis in tumor cells. Together, these proteins provide new molecular handles for the design of therapeutic interventions for CNS injuries and neurodegenerative diseases, as well as possible leads to anticancer strategies. PMID- 11891770 TI - Changes of KCl sensitivity of proliferating neural progenitors during in vitro neurogenesis. AB - The effects of KCl-treatment on the survival and proliferation of NE-4C self renewing neural progenitor cells were investigated during early phases of in vitro induced neurogenesis. NE-4C cells, derived from the anterior brain vesicles of embryonic mouse (E9), divided continuously under non-inducing conditions, but acquired neuronal features within 6 days, if induced by all-trans retinoic acid (RA). During the first 2 days of induction, the cells went on proliferating and did not show signs of morphological differentiation. In this stage, the resting membrane potential of RA-induced cells adopted more negative values in comparison to non-induced ones. Despite the increased membrane polarity and K+ conductance, addition of 20-50 mM KCl failed to elicit inward Na+ currents and did not induce an increase in the intracellular Ca+ level. Long-term treatment with 25 mM KCl, on the other hand, resulted in a selective loss of cells committed to neuronal fate by both decreasing the rate of cell proliferation and increasing the rate of cell death. The data indicate that the viability and proliferation of neural progenitors are influenced by extracellular K+-level in a differentiation stage dependent manner. PMID- 11891769 TI - Fibroblast growth factor-2 converts PACAP growth action on embryonic hindbrain precursors from stimulation to inhibition. AB - Pituitary adenylyl cyclase activating peptide (PACAP) has been shown either to stimulate or to inhibit neural cell proliferation depending on the origin of the cell population. We show here that, depending on the presence or absence of fibroblast growth factor-2 (FGF-2, also called basic FGF), PACAP may either stimulate or inhibit DNA synthesis in neural precursors isolated from embryonic day 10.5 mouse hindbrain. In the absence of FGF-2, PACAP stimulated 3H-thymidine incorporation in a dose-dependent manner. This stimulatory action was unaffected by antagonists of protein kinases A and C but was abolished in the presence of the MEK1/2 antagonist PD98059. In contrast, when FGF-2 was present, PACAP inhibited DNA synthesis. This inhibitory action was insensitive to PD98059 but was fully blocked by the protein kinase A (PKA) inhibitor H89. The differential blockades by MEK1/2 and PKA inhibitors indicate that the FGF-2-induced switch in PACAP action on DNA synthesis was accomplished by a change in PACAP signaling pathways. We hypothesize that the actions of PACAP in the specific parts of the developing nervous system are determined in part by the presence or absence of FGFs and other growth factors. PMID- 11891771 TI - Viral vector-mediated delivery of competing glycosyltransferases modifies epitope expression cell specifically. AB - The glycoconjugate epitopes 3-fucosyl-N-acetyllactosamine (CD15) and sulfoglucuronylcarbohydrate (SGC) mediate cell adhesion events in several systems, and are regulated both spatially and temporally during cerebellar development. In cotransfection studies using COS-1 cells, competition between glycosyltransferases that utilize a common precursor involved in the final synthetic steps of these epitopes, can modulate epitope expression. For example, cotransfection of rat alpha1,3-fucosyltransferase IV (Fuc-TIV) and either rat glucuronic acid transferase P (GlcAT) or pig alpha1,3-galactosyltransferase (GalT) resulted in the dominance of either SGC or GalalphaGal epitope expression, respectively, with blockage of CD15 epitope expression. Viral vectors expressing these glycosyltransferases were used to determine whether competition plays a role in establishing epitope dominance in cerebellar cells, and whether overexpression of competing glycosyltransferases could be used to block epitope expression. Infection of cerebellar astrocytes with viral vectors expressing either Fuc-TIV, or Fuc-TIX, caused dramatic increases in CD15 expression in the presence of continued endogenous SGC epitope expression. Likewise, viral transduction with GalT resulted in GalalphaGal expression without affecting endogenous CD15 or SGC expression. Thus, competition between these enzymes does not appear to play a role in establishing epitope expression in astrocytes, and transduction of these enzymes does not provide a method of blocking the expression of endogenous epitopes. In contrast to what was observed for astrocytes, infection with viral vectors expressing either Fuc-T, GlcAT, or GalT did not result in significant expression of the relevant epitopes (CD15, SGC or GalalphaGal, respectively) on granule neurons. These results suggest a different complement of precursors are present in granule neurons and astrocytes, presumably due to the presence of different complements of glycosyltransferases in these cells. PMID- 11891772 TI - Identification of neuronal cell lineage-specific molecules in the neuronal differentiation of P19 EC cells and mouse central nervous system. AB - P19 embryonic carcinoma (EC) cells are one of the simplest systems for analyzing the neuronal differentiation. To identify the membrane-associated molecules on the neuronal cells involved in the early neuronal differentiation in mice, we generated two monoclonal antibodies, SKY-1 and SKY-2, by immunizing rats with a membrane fraction of the neuronally committed P19 EC cells as an antigen. SKY-1 and SKY-2 recognized the carbohydrate moiety of a 90 kDa protein (RANDAM-1) and the polypeptide core of a 40 kDa protein (RANDAM-2), respectively. In the P19 EC cells, the expression of RANDAM-1 was colocalized to a part of Nestin-positive cells, whereas that of RANDAM-2 was observed in most Nestin-positive cells as well as beta-III-tubulin positive neurons. In the embryonic and adult brain of mice, RANDAM-1 was expressed at embryonic day 8.5 (E8.5), and the localization of antigen was restricted on the neuroepithelium and choroid plexus. The RANDAM-2 expression commenced at E6.0, and the antigen was distributed not only on the neuroepithelium of embryonic brain but on the neurons of adult brain. Collectively, it was concluded that RANDAM-1 is a stage specific antigen to express on the neural stem cells, and RANDAM-2 is constitutively expressed on both the neural stem cells and differentiated neuronal cells in mouse central nervous system (CNS). PMID- 11891773 TI - Pre- and postsynaptic maturation of the neuromuscular junction during neonatal synapse elimination depends on protein kinase C. AB - The distribution of acetylcholine receptors (AChRs) within and around the neuromuscular junction changes dramatically during the first postnatal weeks, a period during which polyneuronal innervation is eliminated. We reported previously that protein kinase C (PKC) activation accelerates postnatal synapse loss. Because of the close relationship between axonal retraction and AChR cluster dispersal, we hypothesize that PKC can modulate morphological maturation changes of the AChR clusters in the postsynaptic membrane during neonatal axonal reduction. We applied substances affecting PKC activity to the neonatal rat levator auris longus muscle in vivo. Muscles were then stained immunohistochemically to detect both AChRs and axons. We found that, during the first postnatal days of normal development, substantial axonal loss preceded the formation of areas in synaptic sites that were free of AChRs, implying that axonal loss could occur independently of changes in AChR cluster organization. Nevertheless, there was a close relationship between axonal loss and AChR organization; PKC modulates both, although differently. Block of PKC activity with calphostin C prevented both AChR loss and axonal loss between postnatal days 4 and 6. PKC may act primarily to influence AChR clusters and not axons, insofar as phorbol ester activation of PKC accelerated changes in receptor aggregates but produced relatively little axon loss. PMID- 11891774 TI - Brain armadillo protein delta-catenin interacts with Abl tyrosine kinase and modulates cellular morphogenesis in response to growth factors. AB - delta-Catenin associates with adhesive junctions and facilitates cellular morphogenesis (Lu et al., 1999). Here we show that delta-catenin colocalizes with actin filaments and Abl tyrosine kinase in the growth cones of cultured hippocampal neurons. PC12 cells induced to express delta-catenin show accelerated neurite extension upon nerve growth factor (NGF) stimulation. STI571, an Abl family kinase inhibitor, further accentuates these stimulatory effects. delta Catenin is a potent substrate for Abl in vitro using an immunocomplex assay and most of the Abl-induced tyrosine phosphorylation within cells is present in the N terminus of delta-catenin. When delta-catenin-expressing epithelial cells are induced to scatter in response to hepatocyte growth factor (HGF), STI571 leads to the rapid redistribution of delta-catenin and changes in cellular morphology. We suggest that delta-catenin is a possible Abl substrate and acts downstream of Abl to orchestrate actin-based cellular morphogenesis. PMID- 11891775 TI - Transient expression of fluorescent tau proteins promotes process formation in PC12 cells: contributions of the tau C-terminus to this process. AB - The neuronal microtubule-associated protein tau promotes microtubule assembly and has been implicated in the development of axonal morphology. In this study, PC12 cells were transiently transfected with constructs coding fusion proteins of human tau with green fluorescent protein (GFP). Expression of tau constructs actively stabilized microtubules. Expression of the C-terminus of tau can mimic this effect in living cells, though to a lesser extent because of the absence of the tau N-terminus. However, tau colocalization with microtubules did not require the presence of the tau N-terminus. Transient expression of tau (including tau24, a four-repeat human tau isoform encoded in 383 residues, and tau23, human fetal tau isoform encoded in 352 residues) stimulated process formation in PC12 cells, and this occurred faster with tau24 than with tau23. The residues (residues 154 172 in tau23) that confer microtubule nucleation activity of tau in vitro are not required for tau-directed process formation. However, when tau induces the formation of cellular processes in response to cortical breakdown by cytochalasin B, residues 154-172 must be present. Thus, it appears that tau may serve to promote cellular process outgrowth in cultured neuronal cells and that C-terminus of tau is essential to this process. PMID- 11891776 TI - Secreted phospholipase A2 potentiates glutamate-induced calcium increase and cell death in primary neuronal cultures. AB - Secreted phospholipases A2 (sPLA2s) modulate neuronal survival and neurotransmitter release. Here we show that sPLA2 (group III) synergistically increases glutamate-induced cell death and intracellular calcium ([Ca2+]i) in cultured primary cortical and hippocampal neurons. Whereas 1 microM glutamate elicited transient [Ca2+]i increases in all neurons that recovered 66% to baseline, 25 ng/ml sPLA2 pretreatment resulted in sustained [Ca2+]i increases, with only 5% recovery. At 250 nM glutamate, 25% of neurons failed to respond, and the average recovery time was 101 +/- 12 sec; sPLA2 increased recovery time to 158 +/- 6 sec, and only 2% of cells failed to respond. Both the noncompetitive N methyl-D-aspartate (NMDA) receptor antagonist MK-801 and the calcium-channel blocker cobalt inhibited this effect. Experiments with the glutamate uptake inhibitor L-trans-pyrollidine-2,4-dicarboxylic acid (2.5 microM) indicated that glutamate uptake sites are not a likely modulation point by sPLA2, whereas arachidonic acid (AA) potentiated calcium responses to glutamate. Thus the enhancement of glutamate-induced [Ca2+]i increases by sPLA2 may be due to modulation at NMDA receptors and/or calcium channels by AA. These results indicate that sPLA2 affects neuronal responses to both nontoxic (0.1-10 microM) and toxic (=25 microM) concentrations of glutamate, implicating this enzyme in neuronal functions in pathology. PMID- 11891777 TI - High-density microarray analysis of hippocampal gene expression following experimental brain injury. AB - Behavioral, biophysical, and pharmacological studies have implicated the hippocampus in the formation and storage of spatial memory. Traumatic brain injury (TBI) often causes spatial memory deficits, which are thought to arise from the death as well as the dysfunction of hippocampal neurons. Cell death and dysfunction are commonly associated with and often caused by altered expression of specific genes. The identification of the genes involved in these processes, as well as those participating in postinjury cellular repair and plasticity, is important for the development of mechanism-based therapies. To monitor the expression levels of a large number of genes and to identify genes not previously implicated in TBI pathophysiology, a high-density oligonucleotide array containing 8,800 genes was interrogated. RNA samples were prepared from ipsilateral hippocampi 3 hr and 24 hr following lateral cortical impact injury and compared to samples from sham-operated controls. Cluster analysis was employed using statistical algorithms to arrange the genes according to similarity in patterns of expression. The study indicates that the genomic response to TBI is complex, affecting approximately 6% (at the time points examined) of the total number of genes examined. The identity of the genes revealed that TBI affects many aspects of cell physiology, including oxidative stress, metabolism, inflammation, structural changes, and cellular signaling. The analysis revealed genes whose expression levels have been reported to be altered in response to injury as well as several genes not previously implicated in TBI pathophysiology. PMID- 11891778 TI - Hypothermia inhibits translocation of CaM kinase II and PKC-alpha, beta, gamma isoforms and fodrin proteolysis in rat brain synaptosome during ischemia reperfusion. AB - To clarify the involvement of intracellular signaling pathway and calpain in the brain injury and its protection by mild hypothermia, immunoblotting analyses were performed in the rat brain after global forebrain ischemia and reperfusion. After 30 min of ischemia followed by 60 min of reperfusion, Ca2+/calmodulin-dependent kinase II (CaM kinase II) and protein kinase C (PKC)-alpha, beta, gamma isoforms translocated to the synaptosomal fraction, while mild hypothermia (32 degrees C) inhibited the translocation. The hypothermia also inhibited fodrin proteolysis caused by ischemia-reperfusion, indicating the inhibition of calpain. These effects of hypothermia may explain the mechanism of the protection against brain ischemia-reperfusion injury through modulating synaptosomal function. PMID- 11891779 TI - Retinoic acid increases proliferation rate of GL-15 glioma cells, involving activation of STAT-3 transcription factor. AB - The molecular mechanisms underlying the heterogeneous effects of retinoic acid (RA) treatment on malignant glioma cells remain poorly understood. In this study, we present the first evidence of a functional role of the signal transduction factors (STATs) in RA-induced proliferation, in a human glioblastoma GL-15 cell line. We first observed that STAT-3 was constitutively activated and present in the GL-15 cell nuclei. We then showed that at low doses (0.01-1 microM) RA increased both the proliferation rate of GL-15 cells and the phosphotyrosine (PY) activation of STAT-3. This RA effect involved transcriptional processes and the transactivation of RA target genes, including RA receptors isoforms RARalpha2, beta2, and -gamma2. At higher concentrations, however, RA (5-10 microM) inhibits GL-15 proliferation, induces apoptosis, and fails to activate STAT-3. An inhibitory effect on GL-15 proliferation was also observed with the synthetic retinoids CD-437 and CD-2325, two structurally related RARgamma agonists, which also fail to activate STAT-3. In addition, the phorbol ester PMA, an inducer of GL-15 differentiation, and staurosporine, a broad inhibitor of protein kinases, abrogate the stimulatory effects of RA at low concentrations. Together these observations suggest that, in GL-15 cells, activation of STAT-3 and cell proliferation share common mechanisms and that STAT transcription factors may be involved in a switch between proliferation, differentiation, and apoptosis. The proliferating effect observed at low doses of RA may be related to the failures in RA efficiency observed in clinical assays in relapsing malignant gliomas. Combining specific inhibitors of tyrosine kinases with RA might optimize the clinical outcome. PMID- 11891780 TI - Effects of cytokine deficiency on chemokine expression in CNS of mice with EAE. AB - Although both cytokines and chemokines have been implicated in the pathogenesis of clinical and histological EAE, their interactions in vivo have not yet been clearly established. To address this issue, we evaluated expression of chemokines and receptors in the CNS of wild-type control and cytokine deficient mice at the peak of EAE induced with MOG-35-55 peptide in CFA. Our results demonstrate that: 1) expression of most chemokines/receptors was drastically inhibited in TNF-alpha deficient mice, and was reflective of delayed onset and reduced severity of EAE; 2) distinct patterns of chemokine expression occurred in various other cytokine knockout mice that did not significantly affect expression of clinical EAE; 3) there was a strong association between expression of MIP-1alpha, MIP-2 and MCP-1 in CNS and overall severity of EAE in wild-type and cytokine knockout mice; and 4) among CNS infiltrating cells at the peak of EAE, macrophages and CD8+ T cells were the primary cellular source of most of the chemokines. Of note, we present evidence that TNF-alpha may be involved in regulating RANTES and MIP-1alpha, and that IL-4 may be involved in regulating MCP-1. Our results not only identify the cellular source of chemokines in CNS, but also implicate MIP-1alpha, MIP-2, and MCP-1 in controlling CNS inflammation and severity of EAE. PMID- 11891781 TI - Effects of chronic nicotine administration on nitric oxide synthase expression and activity in rat brain. AB - Although there is substantial evidence concerning the influence of nicotine on nitric oxide (NO) synthesis in the vascular system, there are fewer studies concerning the central nervous system. Although NO metabolites (nitrates/nitrites) increase in several rat brain regions after chronic injection of nicotine, the cellular origin of this rise in NO levels is not known. The aim of the present work was to assess the effects of repetitive nicotine administration on nitric oxide synthase (NOS) expression and activity in male and female rat brains. To determine levels of nitrate/nitrite, the Griess reaction was carried out in tissue micropunched from the frontal cortex, striatum, and accumbens of both male and female rats untreated (naive) or injected with saline or nicotine (0.4 mg/kg for 15 days). In parallel, coronal sections of fixed brains from equally treated animals were immunostained for neuronal NOS or histochemically labelled for NADPH-diaphorase activity. Nicotine treatment increased NO metabolites significantly in all brain regions compared with naive or saline-treated rats. By contrast, analysis of the planimetric counting of NOS/NADPH-diaphorase-positive neurons failed to demonstrate any significant effect of the nicotine treatment. A significant decrease was observed with both techniques employed in saline-injected female rats compared with naive animals, suggesting a stress response. The mismatch between the biochemical and the histological data after chronic nicotine treatment is discussed. The up regulation of NO sources other than neurons is proposed. PMID- 11891782 TI - Visual responses of neurons in the nucleus of the basal optic root to stationary stimuli in pigeons. AB - The nucleus of the basal optic root of the accessory optic system in pigeons is involved in generating optokinetic nystagmus, which stabilizes object images on the retina by compensatory eye movements. Previous studies have indicated that basal optic neurons are selective for the direction and velocity of motion. The present study shows that these optokinetic cells also respond to stationary stimuli and thereby could be categorized into three groups. The first group of cells (69.1%) responds to stationary gratings orthogonal to the preferred direction but not to gratings parallel to the preferred direction. They do not respond to stationary random-dot patterns without any orientational cues. The second group of cells (7.4%) almost equally discharges a series of bursts in response to stationary gratings with any orientations and to random-dot patterns as well. The third group of cells (23.5%) is responsive to motion but not to stationary gratings and random-dot patterns. The receptive field of basal optic cells is composed of an excitatory field and an inhibitory field, both of which overlap or occupy different regions in the visual field. The aforementioned properties may be attributed to the excitatory receptive field, whereas the inhibitory receptive field is functional when visual stimuli are moving in the direction opposite to the preferred direction of basal optic cells. The functional significance of visual responses of optokinetic neurons to stationary patterns is discussed. PMID- 11891783 TI - The MAGE proteins: emerging roles in cell cycle progression, apoptosis, and neurogenetic disease. AB - Since the identification of the first MAGE gene in 1991, the MAGE family has expanded dramatically, and over 25 MAGE genes have now been identified in humans. The focus of studies on the MAGE proteins has been their potential for cancer immunotherapy, as a result of the finding that peptides derived from MAGE gene products are bound by major histocompatibility complexes and presented on the cell surface of cancer cells. However, the normal physiological role of MAGE proteins has remained a mystery. Recent studies are now beginning to provide insights into MAGE gene function. Necdin acts as a cell cycle regulatory protein and plays a key role in the pathogenesis of Prader-Willi syndrome, a neurogenetic disorder. MAGE-D1, identified as a binding partner for the p75 neurotrophin receptor, the apoptosis inhibitory protein XIAP, and Dlx/MSX homeodomain proteins, blocks cell cycle progression and enhances apoptosis. This review provides an overview of the human MAGE genes and proteins, summarizes recent findings on their cellular roles, and provides a baseline for future studies on this intriguing gene family. PMID- 11891784 TI - Participation of structural microtubule-associated proteins (MAPs) in the development of neuronal polarity. AB - Several lines of evidence have indicated that changes in the structure of neuronal cytoskeleton provide the support for the dramatic morphological changes that occur during neuronal differentiation. It has been proposed that microtubule associated proteins can contribute to the development of this phenomenon by controlling the dynamic properties of microtubules. In this report we have characterized the effect of the combined suppression of MAP1B and tau, and MAP1B and MAP2 on neuronal polarization in cultured hippocampal cells grown on a laminin-containing substrate. We have taken advantage of the use of a mouse line deficient in MAP1B expression obtained by the gene trapping approach. In addition to this engineered mice line we used the antisense oligonucleotide approach to induce the suppression of tau or MAP2, in wild type and MAP1B-deficient neurons. Together these results show a synergistic role for MAP1B/MAP2 and MAP1B/TAU. PMID- 11891785 TI - Calcium/calmodulin-dependent protein kinase IIbeta isoform is expressed in motor neurons during axon outgrowth and is part of slow axonal transport. AB - Previously, we identified calcium/calmodulin-dependent protein kinase IIbeta (CaMKIIbeta) mRNA in spinal motor neurons with 372 bp inserted in what corresponds to the "association" domain of the protein. This was interesting because known additions and deletions to CaMKIIbeta mRNA are usually less than 100 bp in size and found in the "variable" region. Changes in the association domain of CaMKIIbeta could influence substrate specificity, activity or intracellular targeting. We show that three variations of this insert are found in CNS neurons or sciatic motor neurons of Sprague-Dawley rats. We used PCR and nucleic acid sequencing to identify inserts of 114, 243, or 372 bases. We also show that addition of the 372 bases is associated with outgrowth of the axon (the standard CaMKIIbeta downregulates when axon outgrowth occurs). Radiolabeling, immunoblots, and 2D PAGE identified this larger CaMKIIbeta as part of the group of soluble proteins moving at the slowest rate of axonal transport (SCa) in sciatic motor neurons (similar1 mm/day). This group is composed mainly of structural proteins (e.g., tubulin) used to assemble the cytoskeleton of regrowing axons. PMID- 11891786 TI - Developmental changes in the protein composition of sphingolipid- and cholesterol enriched membrane domains of rat cerebellar granule cells. AB - The biological role of cell membrane domains has been investigated in a number of eukariotic cells, but less attention has been paid to the neuron. In the present investigation, we assessed the changes in lipid and protein composition of detergent-resistant membrane fractions prepared from cultured rat cerebellar granule cells, during differentiation and maturation in vitro. At any stage of the cell life, low-density, detergent-resistant fractions, characterised by the specific presence of prion protein, were enriched in glycolipids, cholesterol, and sphingomyelin. The enrichment in sphingomyelin was developmentally regulated, increasing continuously during cell differentiation and maturation. Concerning proteins, domains were enriched in Fyn and TAG-1, which present exclusively within this fraction at any stage of cell culture, and in GAP-43, mainly during the differentiation stage. On the other side, proteins affecting signal transduction and cytoskeleton-related proteins (heterotrimeric G-proteins, protein kinase C, MARCKS, tubulin), were not enriched within detergent-resistant fractions during cell differentiation, but were recovered within this fraction in mature neurons. These results indicate that during different cellular life stages, specific proteins are recruited within detergent-resistant membrane domains of the neuron and suggest their involvement in specific physiological phenomena (differentiation, maturation and/or aging). PMID- 11891787 TI - gamma-Aminobutyric acidA rho receptor subunits in the developing rat hippocampus. AB - The RT-PCR approach was used to estimate the expression of gamma-aminobutyric acid (GABA)(A) rho receptor subunits in the hippocampus of neonatal and adult rats. All three rho subunits were detected at postnatal day (P) 2, the rho3 subunit being expressed at an extremely low level. The rho1 and rho2 products appeared to be developmentally regulated; they were found to be more pronounced in adulthood. In another set of experiments, to correlate gene expression with receptor function, GABA(A) rho subunit mRNAs were detected with single-cell RT PCR in CA3 pyramidal cells (from P3-P4 hippocampal slices), previously characterized with electrophysiological experiments for their bicuculline sensitive or -insensitive responses to GABA. In 6 of 19 cells (31%), pressure application of GABA evoked at -70 mV inward currents that persisted in the presence of 100 microM bicuculline (314 plus minus 129 pA). RT-PCR performed in two of these neurons revealed the presence of rho1 and rho2 subunits, the latter being present with the alpha2 subunit. A rho2 subunit was also found in 1 neuron (among 9) exhibiting a response to GABA, which was completely abolished by bicuculline. This might be due to the lack of putative accessory GABA(A) subunits that can coassemble with rho2 to make functional receptors. Similar experiments from 10 P15 CA3 pyramidal cells failed to reveal any rho1-3 transcripts. However, these neurons abundantly express alpha3 subunits. It is likely that in CA3 pyramidal cells of neonatal and adult hippocampus GABA(A) rho subunits are present but at very low levels of expression. PMID- 11891788 TI - Trafficking of axonal K+ channels: potential role of Hsc70. AB - Voltage-gated potassium ion channels in axons underlie the repolarization phase of the membrane action potential and help to set the resting potential. In addition to being present in the axolemma, they are also found in axoplasm in small vesicles, 30-50 nm in diameter, which may serve as a reserve pool of K+ channel protein (Clay and Kuzirian [2000] J Neurobiol 45:172-184). We have developed a novel technique for extracting these vesicles from axoplasm, which relies on the ability of Texas red to bind to them, thereby reducing their buoyancy so that they are amenable to pelleting by ultracentrifugation (Clay and Kuzirian [2000] J Neurobiol 45:172-184). The mechanism underlying this process may be binding of Texas red to Hsc70, which is primarily a cytosolic protein. However, a small portion of it is located on the surface of vesicles. Kinesin is also on the vesicle surface. This protein is membrane bound in our in vitro vesicle preparation when solutions that do not contain MgATP are added to extruded axoplasm. The addition of MgATP to the solution appears to release a significant amount of kinesin from the vesicles, possibly by the Hsc70-MgATP catalysis mechanism recently proposed by Tsai et al. PMID- 11891789 TI - Ultrastructural localization of neuropeptide Y Y1 receptors in the rat medial nucleus tractus solitarius: relationships with neuropeptide Y or catecholamine neurons. AB - Neuropeptide Y (NPY) Y1 receptor (Y1-R) agonists influence cardiovascular regulation. These actions may involve NPY- and catecholamine-containing neurons in the medial nucleus of the solitary tract (mNTS), at the level of the area postrema. The cellular sites through which Y1-R agonists may interact with NPY and catecholamines in the mNTS, however, are not known. To determine potential sites of action for Y1-R agonists, and their relationship to NPY or catecholamines in the mNTS, we used electron microscopic immunocytochemistry for the detection of sequence-specific antipeptide antisera against Y1-R alone or in combination with antisera against NPY or the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH). Analyses were conducted in the rat mNTS, at the level of the area postrema. Y1-R was found mainly in small unmyelinated axons and axon terminals but also in some somata and dendrites as well as a small number of glia. Within axon terminals, labeling for Y1-R was often present on dense core vesicles and small synaptic vesicles as well as extrasynaptic areas of the plasmalemma. Some Y1-R-labeled terminals also contained NPY or TH, suggesting that agonists of Y1-R may influence the release of NPY or catecholamines in the mNTS. In addition, Y1-R was found in dendrites that received asymmetric excitatory-type synapses from unlabeled axon terminals. Some of these dendrites contained NPY or TH, which indicates that Y1-R may be targeted for functional activation within NPY- or catecholamine-expressing neurons in the mNTS. These results demonstrate that Y1-R is a presynaptic receptor in NPY- or catecholamine containing axon terminals within the mNTS as well as a postsynaptic receptor on NPY- or catecholamine-containing neurons that are contacted by axon terminals that likely contain excitatory amino acid transmitters. Agonists of Y1-R in the mNTS may thus affect cardiovascular regulation by modulating NPY, catecholamine, and excitatory amino acid transmission. PMID- 11891790 TI - Myelin protein zero exists as dimers and tetramers in native membranes of Xenopus laevis peripheral nerve. AB - Protein zero (P0) glycoprotein is the major integral membrane protein of the peripheral nervous system myelin in higher vertebrates. Previous findings indicate the formation of tetrameric assemblies from studies on isolated P0. To determine whether in intact myelin the P0 exists as oligomers, we isolated myelin from sciatic nerve of Xenopus laevis and analyzed it using sodium dodecyl sulfate and urea gel electrophoresis. P0 oligomerization was confirmed using Western blotting, which showed monomeric P0 at approximately 30 kDa and oligomeric P0 at approximately 60 kDa and approximately 120 kDa. A variety of denaturing conditions failed to convert any appreciable amount of oligomer to monomer. Instead, the addition of these denaturants further increased the amount of dimer and tetramer while decreasing the amount of monomer. Native gels showed dimeric P0 without the appearance of monomer or tetramer, suggesting that dimeric P0, the most prominent form of the protein, is the most stable and likely occurs in the native myelin membrane array. PMID- 11891791 TI - NF-kappaB and IkappaBalpha expression following traumatic brain injury to the immature rat brain. AB - NF-kappaB is one of the most important modulators of stress and inflammatory gene expression in the nervous system. In the adult brain, NF-kappaB upregulation has been demonstrated in neurons and glial cells in response to experimental injury and neuropathological disorders, where it has been related to both neurodegenerative and neuroprotective activities. Accordingly, the aim of this study was to evaluate the cellular and temporal patterns of NF-kappaB activation and the expression of its endogenous inhibitor IkappaBalpha following traumatic brain injury (TBI) during the early postnatal weeks, when the brain presents elevated levels of plasticity and neuroprotection. Our results showed that cortical trauma to the 9-day-old rat brain induced a very fast upregulation of NF kappaB, which was maximal within the first 24 hours after injury. NF-kappaB was mainly observed in neuronal cells of the degenerating cortex as well as in astrocytes located in the corpus callosum adjacent to the injury, where a pulse like pattern of microglial NF-kappaB activation was also found. In addition, astrocytes of the corpus callosum, and microglial cells to a lower extent, also showed de novo expression of IkappaBalpha within the time of NF-kappaB activation. This study suggests an important role of NF-kappaB activation in the early mechanisms of neuronal death or survival, as well as in the development of the glial and inflammatory responses following traumatic injury to the immature rat brain. PMID- 11891792 TI - N-methyl-D-aspartate receptors: transient loss of NR1/NR2A/NR2B subunits after traumatic brain injury in a rodent model. AB - Hippocampal N-methyl-D-aspartate (NMDA) receptor subunits, by virtue of their involvement in excitotoxic injury as well as memory association, may play an important role in the pathophysiologic mechanisms of traumatic brain injury (TBI). In this study, temporal changes in NMDA receptor subunit (NR1, NR2A, and NR2B) levels in rat hippocampus after TBI were investigated by Western blot and mRNA expression levels by RT-PCR methods. Sprague-Dawley rats (250-350 g) were employed, and a controlled cortical impact injury device was used to produce the TBI in rodents. At different postinjury time points (2, 6, 12, 24, and 48 hr), the rat hippocampi were dissected out for protein and RNA preparation. Western blot analysis revealed significant decreases of NR1, NR2A, and NR2B subunit proteins at 6 and 12 hr postinjury in rat hippocampus. Complete recovery of NR1, NR2A, and NR2B subunit protein to the levels of sham controls was observed at 24 hr postinjury. However, RT-PCR analysis did not show any significant change in the mRNA levels at 2, 6, and 12 hr postinjury in comparison with sham controls, suggesting nontranscriptional change in the levels of these subunits. Thus, TBI can produce transient degradation of NMDA receptor subunits in the hippocampus, which might contribute to temporary memory impairment after injury. PMID- 11891793 TI - Peptidyl alpha-keto amide inhibitor of calpain blocks excitotoxic damage without affecting signal transduction events. AB - The cysteine protease calpain is activated by calcium and has a wide range of substrates. Calpain-mediated cellular damage is associated with many neuropathologies, and calpain also plays a role in signal transduction events that are essential for cell maintenance, including the activation of important kinases and transcription factors. In the present study, the hippocampal slice culture was used as a model of excitotoxicity to test whether the neuroprotection elicited by selective calpain inhibition is associated with changes in cell signaling. Peptidyl alpha-keto amide and alpha-keto acid inhibitors reduced both calpain-mediated cytoskeletal damage and the concomitant synaptic deterioration resulting from an N-methyl-D-aspartate exposure. The alpha-keto amide CX295 was protective when infused into slice cultures before or after the excitotoxic episode. The slices protected with CX295 exhibited normal activation levels of mitogen-activated protein kinase and the transcription factor nuclear factor kappaB. Thus, selective inhibition of calpain provides neuroprotection without influencing critical signaling pathways. PMID- 11891794 TI - Erythropoietin protects neurons against chemical hypoxia and cerebral ischemic injury by up-regulating Bcl-xL expression. AB - Erythropoietin (EPO) promotes neuronal survival after cerebral ischemia in vivo and after hypoxia in vitro. However, the mechanisms underlying the protective effects of EPO on ischemic/hypoxic neurons are not fully understood. The present in vitro experiments showed that EPO attenuated neuronal damage caused by chemical hypoxia at lower extracellular concentrations (10(- 4)-10(-2) U/ml) than were previously considered. Moreover, EPO at a concentration of 10(-3) U/ml up regulated Bcl-xL mRNA and protein expressions in cultured neurons. Subsequent in vivo study focused on whether EPO rescued hippocampal CA1 neurons from lethal ischemic damage and up-regulated the expressions of Bcl-xL mRNA and protein in the hippocampal CA1 field of ischemic gerbils. EPO was infused into the cerebroventricles of gerbils immediately after 3 min of ischemia for 28 days. Infusion of EPO at a dose of 5 U/day prevented the occurrence of ischemia-induced learning disability. Subsequent light microscopic examinations showed that pyramidal neurons in the hippocampal CA1 field were significantly more numerous in ischemic gerbils infused with EPO (5 U/day) than in those receiving vehicle infusion. The same dose of EPO infusion caused significantly more intense expressions of Bcl-xL mRNA and protein in the hippocampal CA1 field of ischemic gerbils than did vehicle infusion. These findings suggest that EPO prevents delayed neuronal death in the hippocampal CA1 field, possibly through up regulation of Bcl-xL, which is known to facilitate neuron survival. PMID- 11891795 TI - Baclofen is neuroprotective and prevents loss of calcium/calmodulin-dependent protein kinase II immunoreactivity in the ischemic gerbil hippocampus. AB - Excessive release of glutamate during transient cerebral ischemia initiates a cascade of events that leads to the delayed and selective death of neurons located in the hippocampus. Activity of calcium calmodulin kinase II (CaM kinase), a protein kinase critical to neuronal functioning, disappears following ischemia. The in vivo link between glutamate excitoxicity and alterations in CaM kinase activity has not been extensively studied. Baclofen, a selective gamma aminobutyric acid (GABA)(B) receptor agonist, has been shown to inhibit glutamate release. The present study evaluated the neuroprotective efficacy of this compound and assessed early changes in hippocampal-dependent behaviors and CaM kinase immunoreactivity following transient cerebral ischemia. Baclofen (50 mg/kg) prevented both the loss of hippocampal CA1 pyramidal cells and the reduction in hippocampal CaM kinase immunoreactivity observed in control animals following ischemic insult. Cerebral ischemia produced a significant increase in working memory errors; however, baclofen failed to attenuate this memory deficit. Results confirm that baclofen is neuroprotective and support a link between glutamate excitotoxicity and reductions in CaM kinase immunoreactivity. PMID- 11891796 TI - Differential expression of the "C" and "T" alleles of the 5-HT2A receptor gene in the temporal cortex of normal individuals and schizophrenics. AB - A genetic association between schizophrenia and a silent C/T(102) polymorphism in the 5-HT2A receptor gene (5-HT2AR) has been previously reported; however, the mechanisms underlying this association remain unknown. Here we developed an improved quantitative assay for measurements of allele ratios, which revealed that the expression of allele "C" in the temporal cortex of normal heterozygous individuals was significantly lower than the expression of allele "T" (allele "C" to allele "T" ratio of approximately 0.8, P < 0.0001). Confirming decreased expression of allele "C," total levels of 5-HT2AR mRNA and protein in normal individuals with the C/C genotype were lower than in individuals with the T/T genotype. Similarly to normal individuals, allele "C" to allele "T" ratio in heterozygous schizophrenics was reduced (approximately 0.8, P < 0001). This ratio was independent of neuroleptic treatment history. By contrast, total levels of 5 HT2AR mRNA correlated inversely with neuroleptic free interval prior to death (r = -0.67, P < 0.001) suggesting a reversible neuroleptic effect. Total levels 5 HT2AR mRNA in neuroleptic free (> 26 weeks) schizophrenics (n = 11) were significantly lower than in controls (P = 0.03). The data suggest that increased prevalence of allele "C" among schizophrenics may be due to intrinsically low expression of this allele, which may contribute to a deficit in 5-HT2AR expression in some schizophrenics. PMID- 11891797 TI - Initiation and development of experimental autoimmune neuritis in Lewis rats is independent of the cytotoxic capacity of NKR-P1A+ cells. AB - Natural killer (NK) cells are implicated in T cell-mediated autoimmune diseases. Experimental autoimmune neuritis (EAN) is a CD4+ T cell-mediated animal model of the Guillain-Barre syndrome in human. The role of NK cells in the initiation and development of EAN remains unclear. In the present study, we demonstrate that anti-NKR-P1A monoclonal antibody (mAb) treatment in vivo did not affect the initiation and development of clinical EAN in Lewis rats induced by immunization with peripheral nerve myelin P0 protein peptide 180-199 and Freund's complete adjuvant, as well as the proportion of NKR-P1A+ cells (including NK cells and NKT cells) in the spleen. Furthermore, inflammatory cell infiltrations and demyelination in the peripheral nervous system (PNS) and in vitro P0 peptide 180 199-specific splenocyte proliferation were not different in anti-NKR-P1A mAb treated rats compared to the control antibody-treated rats. The cytotoxic activity of NKR-P1A+ cells, determined by NK cell-sensitive K562 cells as target cells, decreased markedly in anti-NKR-P1A mAb-treated rats, suggesting that decrease of the cytotoxic activities of NKR-P1A+ cells is not sufficient to alter clinical EAN, although NKR-P1A+ cells may participate in the pathogenesis of T cell-mediated autoimmune diseases, such as EAN, by the mechanisms that involve the release of cytokines. PMID- 11891798 TI - Role of CB1 and CB2 receptors in the inhibitory effects of cannabinoids on lipopolysaccharide-induced nitric oxide release in astrocyte cultures. AB - The purpose of this study was to investigate the role of the central cannabinoid receptor (CB(1)) in mediating the actions of the endogenous cannabinoid agonist anandamide and the synthetic cannabinoid CP-55940. Activation of primary mouse astrocyte cultures by exposure to bacterial lipopolysaccharide (LPS) caused a marked (approximately tenfold) increase in nitric oxide (NO) release. Coincubation with the cannabinoid agonists anandamide or CP-55940 markedly inhibited release of NO (-12% to -55%). This effect was abolished by SR-141716A (1 microM), a CB1 receptor antagonist. SR-141716A alone also significantly increased NO release in response to LPS, suggesting that endogenous cannabinoids modify inflammatory responses. In contrast, coincubation with the CB2 receptor antagonist SR-144528 (1 microM) abolished the inhibitory effects of the endogenous cannabinoid anandamide on LPS-induced NO release, although this may reflect nonspecific effects of this ligand or cannabinoid actions through atypical receptors of anandamide. We also showed that endogenous or synthetic cannabinoids inhibit LPS-induced inducible NO synthase expression (mRNA and protein) in astrocyte cultures. These results indicate that CB1 receptors may promote antiinflammatory responses in astrocytes. PMID- 11891799 TI - Involvement of caspase-3 and p38 mitogen-activated protein kinase in cobalt chloride-induced apoptosis in PC12 cells. AB - Our previous study showed that cobalt chloride (CoCl2) could induce PC12 cell apoptosis and that the CoCl2-treated PC12 cells may serve as a simple in vitro model for the study of the mechanism of hypoxia-linked neuronal disorders. The aim of this study is to elucidate the mechanism of CoCl2-induced apoptosis in PC12 cells. Caspases are known to be involved in the apoptosis induced by various stimuli in many cell types. To investigate the involvement of caspases in CoCl2 induced apoptosis in PC12 cells, we generated PC12 cells that stably express the viral caspases inhibitor gene p35 and analyzed the effect of p35 on the process of apoptosis induced by CoCl2. We also examined the effect of cell-permeable peptide inhibitors of caspases. The results showed that the baculovirus p35 gene and the general caspases inhibitor Z-VAD-FMK significantly block apoptosis induced by CoCl2, confirming that caspase is involved in CoCl2-induced apoptosis. Further investigation showed that in this process the caspase-3-like activity is increased, as indicated by the cells' ability to cleave the fluorogenic peptide substrate Ac-Asp-Glu-Val-Asp-7-AMC and to degrade the DNA-repairing enzyme poly (ADP-ribose) polymerase (PARP), an endogenous caspase-3 substrate. At the same time, caspase-3-specific inhibitors, namely, the peptide Ac-DEVD-CHO, Ac-DEVD FMK, partially inhibit CoCl2-induced apoptosis. These findings suggested that caspase-3 or caspase-3-like proteases are involved in the apoptosis induced by CoCl2 in PC12 cells. Additionally, we have observed that another apoptotic marker, p38 mitogen-activated protein kinase (MAPK), is significantly activated in this process in a time-dependent manner and that a selective p38 MAPK inhibitor, SB203580, partially inhibits this cell death. The addition of SB203580 also partially suppresses caspase-3-like activity. All these results confirm that the CoCl2-treated PC12 cell is a useful in vitro model with which to study hypoxia-linked neuronal disorders. Furthermore, the results showing that the baculovirus p35 gene and caspase inhibitors possess a remarkable ability to rescue PC12 cells from CoCl2-induced cell death may have implications for future neuroprotective therapeutic approaches for the hypoxia-associated disorders. PMID- 11891800 TI - Cerebrospinal fluid neurofilament and glial fibrillary acidic protein in patients with cerebral vasculitis. AB - Few diseases in clinical medicine cause as much diagnostic consternation as central nervous system (CNS) vasculitis because of its varying modes of presentation and frequently overlapping clinical and pathological features. There are no pathognomonic clinical or laboratory findings. The purpose of the present retrospective study was to validate the use of the light subunit of neurofilament triplet protein (NFL) and glial fibrillary acidic protein (GFAP) as markers of CNS tissue damage for patients with systemic or isolated CNS vasculitis. Levels of cerebrospinal fluid (CSF) NFL and GFAP were measured using ELISAs. Both CSF NFL and CSF GFAP concentrations were significantly higher in a patient group diagnosed with CNS vasculitis (P < 0.01 and P < 0.05, respectively) than in a patient group for whom CNS vasculitis was excluded. In the future, analysis of CSF NFL in particular, but also GFAP, may be a useful complement in the difficult clinical task of diagnosing CNS vasculitis. PMID- 11891801 TI - Soluble angiogenic factors: implications for chronic myeloproliferative disorders. AB - The role of angiogenesis for the progressive growth and metastatic process of tumours is well established. What is not clear, though, is the clinical prognostic significance of the angiogenic factors in malignant haematological diseases. In this study, we have assessed the plasma and serum levels of two major angiogenic factors, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF) in 55 patients affected by chronic myeloproliferative disorders (CMD). This series included 25 patients with essential thrombocythemia (ET), 10 patients with chronic myelocytic leukaemia (CML), 14 patients with polycythemia vera (PV), and 6 patients with primary myelofibrosis (MF), and they were compared to 20 healthy control subjects. In all patients the plasma VEGF concentration was significantly increased to the healthy control group (P < 0.004). The highest concentrations were found in the patients with ET (178.25 +/- 125.22 pg/ml). The VEGF levels were significantly higher in CMD patients with vascular complications than those in CMD patients without complications (P < 0.01). The b-FGF serum levels also appeared to be significantly higher in almost all the CMD patients compared to the control group (P < 0.07). A significant correlation was found between the VEGF levels and the platelet count in the ET patients and the spleen index in the CML patients. VEGF level, in this study, is associated with increased risk of thrombotic complications. There is evidence of increased levels of soluble angiogenic factors in malignant haematological disorders, but their contribution to the progression of diseases is yet unclear. PMID- 11891803 TI - Lymphomas of mucosal-associated lymphoid tissue in common variable immunodeficiency. AB - Common variable immunodeficiency (CVID) is a primary immunodeficiency disease characterized by low serum immunoglobulins IgG, IgA, and usually IgM. The central immune deficiency is impaired secretion of immunoglobulins and lack of antibody production; however, T cell dysfunction and a variety of inflammatory complications suggest global immune dysregulation. A number of reports have documented the association of primary immunodeficiency diseases with the development of non-Hodgkin's lymphoma (NHL). In CVID, the risk has been estimated to lie between 1.4% and 7%. As for NHL arising in other immunodeficiency states, the lymphomas in CVID are extranodal and are usually B cell in type. Of 22 B cell lymphomas that have appeared over a period of 25 years in a cohort of subjects with CVID, five lymphomas, appearing in more recently studied subjects, that arose in mucosal sites would be classified as mucosa-associated lymphoid tissue (MALT) lymphomas. MALT lymphomas are low-grade B cell lymphomas that result from a proliferation of neoplastic marginal-zone related cells of lymphoid tissue and tend to occur in organs that have acquired lymphoid tissue due to long-term infectious or autoimmune stimulation. Lymphomas of this kind have not been described in patients with congenital immunodeficiency, although chronic mucosal antigen stimulation is an integral part of these immune deficiency states. PMID- 11891802 TI - Cellular location of proteins related to iron absorption and transport. AB - K562 erythroleukemia cells and IEC6 rat cells were examined using confocal microscopy and antibodies raised against DMT-1 (Nramp-2, DCT-1), transferrin receptor (CD71), beta(3) integrin (CD61), mobilferrin (calreticulin), and Hephaestin. The cellular location of each of these proteins was identified by immunofluorescence in both saponin-permeabilized and non-permeabilized cells. Fluorescent reactivity was observed on or near the cell surface of each of these proteins, suggesting that they might participate in surface membrane transport of iron. Fluorescence was observed in the region of the cytoplasm with each antibody to include beta(3) integrin and transferrin receptor. It was pronounced in cells incubated with mobilferrin, Hephaestin, and DMT-1 antibodies. Speckled nuclear fluorescence was observed in cells incubated with anti-DMT-1. While these observations are descriptive, they demonstrate that there are significant concentrations of DMT-1, mobilferrin, and Hephaestin in the cytoplasmic region of cells. This suggests that there may be intracellular roles for these proteins in addition to their serving to transit iron across the cell surface membrane. PMID- 11891804 TI - Treatment of children with Langerhans cell histiocytosis with 2 chlorodeoxyadenosine. AB - Langerhans cell histiocytosis (LCH) is a disorder characterized by proliferation of activated Langerhans cells. Immune dysregulation is believed to be part of the pathogenesis. Although current therapies are very effective at inducing remission, multiple recurrences and long-term sequelae are common for patients with low-risk disease, and a significant proportion of young patients die of their disease. More effective therapies based on the pathogenesis of LCH are needed. We investigated the use of 2-chloro-deoxyadenosine (2-CdA), a purine analogue with an antiproliferative effect on histiocytes and lymphocytes, in patients with recurrent or high-risk LCH. Six patients with recurrent LCH received 2-CdA (5-7 mg/m(2)/day for 5 days, repeated every 21-28 days). All patients achieved remission. With a median follow-up of 15 months (range, 3-25 months), 5 patients remain in remission. A patient with multisystem disease who recurred after 13 months, achieved a second remission with 2-CdA. Hematologic toxicity was minimal, and no infectious complications were documented. 2-CdA is among the most effective drugs for the treatment of LCH, and this is probably due to both its anti-proliferative and immunomodulatory effects. 2-CdA needs to be considered for the treatment of recurrent LCH. However, its incorporation into front-line treatment of patients with multi-system LCH needs further study. PMID- 11891805 TI - Frequent occurrence of anticardiolipin antibodies, Factor V Leiden mutation, and perturbed endothelial function in chronic myeloproliferative disorders. AB - Chronic myeloproliferative disorders (MPD) are characterized by a high incidence of thrombohaemorrhagic complications, possibly related to platelet abnormalities and disturbances of the coagulation system. In an attempt to define abnormalities in coagulation and fibrinolysis, we investigated risk markers for venous thromboembolism, fibrinolytic, and haemostatic system activation markers and antiphospholipid antibodies in blood samples from 50 MPD patients and 30 controls. Compared with controls median levels of protein S free and protein C were significantly decreased in the patients (0.27 vs. 0.38 arbitrary units; P < 0.001 and 0.86 vs. 0.99 arbitrary units; P < 0.001, respectively), and activated partial thromboplastin time was significantly prolonged in patients (33 vs. 27 sec; P < 0.001). No differences were observed in levels of antithrombin, thrombin antithrombin complex, and fibrin D-dimer. In patients the median value of thrombomodulin was significantly increased indicating perturbed endothelial function. Anticardiolipin antibodies of IgM subtype (median 38 U/mL, 33-99) were detected in 11 patients (22%) and only in one control (3%) (P < 0.021; patients vs. controls). Seven patients were heterozygous for the Factor V Leiden, one patient was heterozygous for the prothrombin G20210A mutation, and one patient was homozygous for the Factor V Leiden mutation. Among controls, two were heterozygous for the Factor V Leiden mutation. Comparing the allele frequency of the Factor V Leiden mutation in patients with MPD and the background population disclosed a significantly increased allele prevalence of the Factor V Leiden mutation in the patients (9% vs. 3.4%; P = 0.003). Alterations in the level of anticoagulant proteins, disturbances of endothelial cell function, and the presence of cardiolipin antibodies and Factor V Leiden mutation may increase the cumulative thrombotic risk in MPD besides the risk imposed by platelet activation. PMID- 11891806 TI - Platelet activation and hypercoagulability following treatment with porcine factor VIII (HYATE:C). AB - Activation of platelets and coagulation in vivo was studied in nine patients with hemophilia A and inhibitors to human Factor VIII, prior to and following treatment with porcine Factor VIII (PFVIII; HYATE:C). In addition, six hemophiliac patients were similarly studied after treatment with recombinant Factor VIII (rFVIII). Platelet activation was also examined in vitro using porcine von Willebrand factor (PvWF)-enriched and PvWF-depleted fractions obtained by fractionation of PFVIII. Coagulation was assessed by measuring the concentrations of plasma prothrombin fragment 1+2 concentrations (prothrombinase generation) and Factor Xa-ATIII. Patients treated with PFVIII had significantly increased numbers of circulating platelets expressing CD62 and CD63 (markers of platelet activation) and annexin V (marker of platelet procoagulant activity) compared to patients treated with rFVIII; the former patients also demonstrated an increase in plasma coagulability after therapy. In in vitro experiments it was observed that the platelet-activating and procoagulant capacity of PFVIII resided in the PvWF-enriched fraction, and the same was true for the plasma hypercoagulability following exposure of platelets to PFVIII. These results support the hypothesis that PFVIII-induced platelet activation provides a mechanism for enhancing hemostasis, separate from, and additional to, that due to increased circulating Factor VIII, and it is due to residual PvWF in the PFVIII preparation. PMID- 11891807 TI - CD30 and the NPM-ALK fusion protein (p80) are differentially expressed between peripheral blood and bone marrow in primary small cell variant of anaplastic large cell lymphoma. AB - The t(2;5)(p23;q35) translocation results in the formation of a unique chimeric NPM-ALK protein (p80). Expression of this protein is considered to be one of the clinical features of anaplastic large cell lymphoma (ALCL). Recently recognized as one clinical subtype of ALCL, the small cell variant is prone to have a leukemic presentation. Although the small cell variant has been recognized as a subtype of ALCL, the clinical properties of this subtype, especially the immunophenotype of lymphoma cells in peripheral blood, have not yet been fully described. This report shows that neither CD30 nor p80 is detected by immunostaining in the predominant small cell malignant clone and also in large lymphoma cells in peripheral blood, while large cells and occasionally observed small cells in bone marrow were found to be positive for CD30 and p80. Our findings suggest that differential expression of CD30 and p80 between peripheral blood and bone marrow lymphoma cells is a property of the small cell variant of ALCL. PMID- 11891808 TI - Massive splenic infarction in Saudi patients with sickle cell anemia: a unique manifestation. AB - Splenic infarcts are common in patients with sickle cell anemia (SCA), but these are usually small and repetitive, leading ultimately to autosplenectomy. Massive splenic infarcts on the other hand are extremely rare. This is a report of our experience with 8 (4 males and 4 females) cases of massive splenic infarction in patients with SCA. Their ages ranged from 16 to 36 years (mean 22 years). Three presented with left upper quadrant abdominal pain and massive splenic infarction on admission, while the other 5 developed massive splenic infarction while in hospital. In 5 the precipitating factors were high altitude, postoperative, postpartum, salmonella septicemia, and strenuous exercise in one each, while the remaining 3 had severe generalized vasoocclusive crises. Although both ultrasound and CT scan of the abdomen were of diagnostic value, we found CT scan more accurate in delineating the size of infarction. All our patients were managed conservatively with I.V. fluids, analgesia, and blood transfusion when necessary. Diagnostic aspiration under ultrasound guidance was necessary in two patients to differentiate between massive splenic infarction and splenic abscess. Two patients required splenectomy during the same admission because of suspicion of secondary infection and abscess formation, while a third patient had splenectomy 2 months after the attack because of persistent left upper quadrant abdominal pain. In all the 3 histology of the spleen showed congestive splenomegaly with massive infarction. All of our patients survived. Two patients subsequently developed autosplenectomy while the remaining 3 continue to have persistent but asymptomatic splenomegaly. Massive splenic infarction is a rare and unique complication of SCA in the Eastern Province of Saudi Arabia, and for early diagnosis and treatment, physicians caring for these patients should be aware of such a complication. PMID- 11891809 TI - Piebaldism associated with congenital dyserythropoietic anemia type II (HEMPAS). AB - Congenital dyserythropoietic anemias (CDAs) are a group of relatively rare inherited anemias. They are characterized by ineffective erythropoiesis and classified as three major groups and a number of variants. CDA type II, also known as hereditary erythroblastic multinuclearity with a positive acidified serum test (HEMPAS), is the most frequent one. A number of associations with CDA II have been reported, although each described only one or a few patients. Here we presented a piebald woman with vaginal atresia who was tested for anemia and diagnosed as CDA type II. Piebaldism and anemia association were previously described in the mouse. Our case was the first that shows the features of both piebaldism and CDA in the same patient. This association may suggest a stem cell defect to cause both hematopoietic and cutaneous manifestations. PMID- 11891810 TI - New alpha 2 globin chain variant with low oxygen affinity affecting the N terminal residue and leading to N-acetylation [Hb Lyon-Bron alpha 1(NA1)Val --> Ac-Ala]. AB - Hemoglobin Lyon-Bron was found in two members of a family of German ascent presenting with a moderate normocytic anemia. In this alpha 2 globin variant, the N-terminal valine of the chain was replaced by an alanine. Electrospray mass spectrometry of the alpha chain showed that, as normally, the initiator methionine was cleaved during globin processing but that the N alpha-terminal group was totally acetylated. This resulted in structural modifications of a region crucial for oxygen binding. As a consequence, hemoglobin Lyon-Bron displayed both a reduced chloride effect and a decreased oxygen affinity, this last point explaining the apparent anemia. PMID- 11891811 TI - Effective hemostasis with rFVIIa treatment in two patients with severe thrombocytopenia and life-threatening hemorrhage. AB - We report two patients with severe thrombocytopenia and life-threatening bleeding that were successfully managed with recombinant activated factor VII (rFVIIa). The first was a 75-year-old male with Waldenstrom's macroglobulinemia. During a therapeutic course with fludarabine, he developed severe autoimmune thrombocytopenia resistant to conventional treatment, followed by persistent uncontrollable nasal bleeding. Platelet transfusions failed to increase the platelet count and control the hemorrhage. When hemoglobin levels fell below 8.5 g/dL and the patient's clinical condition got much worse, a single dose of 4.8 mg rFVIIa (90 microg/kg) was given as an i.v. bolus. Ten minutes after the rFVIIa injection, nasal bleeding stopped, the patient's clinical condition progressively improved, and splenectomy could be carried out uneventfully 2 days later. The second patient, a 52-year-old female, was under treatment for pre-B lymphoblastic leukemia. She developed severe thrombocytopenia, secondary to chemotherapy, complicated by massive gastrointestinal bleeding. Despite intensive treatment with platelet transfusions, hemorrhage continued and her condition deteriorated rapidly. She was then given an i.v. bolus injection of 4.8 mg rFVIIa, which resulted in cessation of hemorrhage and dramatic improvement of her clinical status. No adverse effects from the treatment with rFVIIa were observed. In conclusion, rFVIIa appears to be an attractive alternative for controlling hemorrhage in patients with severe thrombocytopenia, especially when platelet transfusions are unavailable or ineffective. PMID- 11891812 TI - Severe hypophosphatemia: a rare cause of intravascular hemolysis. AB - A 3-year-old child presented with severe hyperphosphatemia (phosphate 45 mg/dL) secondary to chronic enema use. Following aggressive correction of the hyperphosphatemia, hypophosphatemia ensued (phosphate 1.7 mg/dL). Concurrently, the patient developed severe intravascular hemolysis and RBC morphologic defects. The hemolysis and morphologic defects corrected with return to normal serum phosphate levels. Severe hypophosphatemia is a rare cause of intravascular hemolysis. PMID- 11891813 TI - Anti-D (WinRho SD) treatment of children with chronic autoimmune thrombocytopenic purpura stimulates transient cytokine/chemokine production. AB - Intravenous anti-D is often used in the treatment of autoimmune thrombocytopenic purpura (AITP), but little is known about its mechanisms of action. To investigate anti-D's potential in vivo mechanism(s) of action, a small group (N = 7) of children with chronic AITP was studied. The children initially received either 25 or 50 microg/kg of WinRho-SD in a four-cycle cross-over trial, and peripheral blood samples from the first and third cycles were assessed for cytokine levels at pre-treatment, 3 hr, 1 day, and 8 days post-treatment. Results showed that platelet counts significantly increased in all the children by day 8 post-treatment. Analysis of serum by ELISA showed that there was a significant but transient rise in both pro- and anti-inflammatory cytokine/chemokine levels (e.g., IL1RA, IL6, GM-CSF, MCP-1 alpha, TNF-alpha and MCP-1) by 3 hr post treatment in both cycles which returned to baseline levels by 8 days post treatment. These results suggest that anti-D administration may initially activate the RES in the form of cytokine/chemokine secretion, which is subsequently followed by an increase in platelet counts. It is possible that the induced cytokine/chemokine storm may have an effect on several physiological processes such as those mediating either adverse effects or potentially RES phagocytic activity. PMID- 11891814 TI - Management of a patient with HIV infection-induced anemia and thrombocytopenia who presented with thrombotic thrombocytopenic purpura. AB - A 32-year-old male presented with fever, mental status changes, renal dysfunction, cytopenias and hemolysis. His platelet count was 14,000/microL, hemoglobin 5.7 g/dL and LDH 2,636 U/L. He was diagnosed with thrombotic thrombocytopenic purpura (TTP) and also found to be HIV positive on admission. TTP was confirmed by a low von Willebrand factor-cleaving protease level, the gold standard test for TTP, which was 10-15%. No protease-specific antibody was detected. Treatment of this patient consisted of 23 plasmapheresis procedures and trials of vincristine and dextran-70. Despite therapy, the patient remained anemic and thrombocytopenic, though his mental status and renal abnormalities improved. Highly active anti-retroviral therapy (HAART) consisting of efavirenz, 3TC, and d4T was started. Only after plasma exchanges were discontinued and HAART was instituted did the cytopenias resolve. He continued to improve following discharge, and platelet count was 206,000/microL and hemoglobin, 12.5 g/dL one month after the initiation of HAART. PMID- 11891815 TI - Successful treatment of steroid and cyclophosphamide-resistant hemolysis in chronic lymphocytic leukemia with rituximab. PMID- 11891816 TI - Severe anemia and marrow plasmacytosis as presentation of Sjogren's syndrome. PMID- 11891817 TI - Pregnancy and abortion in women with essential thrombocythemia. PMID- 11891818 TI - Comprehensive cytogenetic study by spectral karyotyping on blastic natural killer cell leukemia/lymphoma. PMID- 11891820 TI - Is homocysteine a causal and treatable risk factor for vascular diseases of the brain (cognitive impairment and stroke)? PMID- 11891821 TI - Memory decline: the boundary between aging and age-related disease. PMID- 11891822 TI - Homocysteine, silent brain infarcts, and white matter lesions: The Rotterdam Scan Study. AB - Silent brain infarcts and white matter lesions are frequently seen on magnetic resonance imaging in healthy elderly people and both are associated with an increased risk of stroke and dementia. Plasma total homocysteine may be a potentially modifiable risk factor for stroke and dementia. We examined whether elevated total homocysteine levels are associated with silent brain infarcts and white matter lesions. The Rotterdam Scan Study is a population-based study of 1,077 people aged 60 to 90 years who had cerebral magnetic resonance imaging. The cross-sectional relation of total homocysteine with silent infarcts and white matter lesions was analyzed with adjustment for cardiovascular risk factors. The mean plasma total homocysteine level was 11.5 micromol/l (standard deviation 4.1). The risk of silent brain infarcts increased with increasing total homocysteine levels (odds ratio 1.24/standard deviation increase, 95% confidence interval 1.06-1.45). The severity of periventricular white matter lesions and extent of subcortical white matter lesions were also significantly associated with total homocysteine levels, even after excluding those with silent brain infarcts. The overall risk of having either a silent brain infarct or severe white matter lesions was strongly associated with total homocysteine levels (odds ratio 1.35/standard deviation increase, 95% confidence interval 1.16-1.58). We concluded that total homocysteine levels are associated with silent brain infarcts and white matter lesions independent of each other and of other cardiovascular risk factors. PMID- 11891823 TI - Imaging hippocampal function across the human life span: is memory decline normal or not? AB - Memory function commonly declines in later life. Whether memory decline represents a disease process or whether it is part of normal aging remains unknown. Here we answer this question by assessing the function of multiple subregions that make up the hippocampal circuit across the human life span. A newly developed MRI approach--designed to detect functional changes in individual hippocampal subregions--was used to assess the hippocampal circuit in 70 subjects between 20 and 88 years of age. Using strict parametric criteria, analysis revealed that function in two hippocampal subregions--the subiculum and the dentate gyrus--decline normally with age. In contrast, function in the entorhinal cortex declines pathologically. Single-subject analysis revealed that hippocampal dysfunction, found in 60% of elders was selectively correlated with memory decline. These results show that memory decline is caused by different mechanisms and suggests how memory decline should be approached clinically. PMID- 11891824 TI - A new locus for Parkinson's disease (PARK8) maps to chromosome 12p11.2-q13.1. AB - We performed genomewide linkage analysis of a Japanese family with autosomal dominant parkinsonism, which exhibits clinical features compatible with those of common Parkinson's disease. Parametric two-point linkage analysis yielded a highest log odds (LOD) score of 4.32 at D12S345 (12p11.21). Parametric multipoint linkage analysis of the 13.6cM interval around this marker yielded LOD scores almost uniformly of >4.0 with a Z(max) of 4.71 at D12S85 (12q12). Haplotype analysis detected two obligate recombination events at D12S1631 and D12S339 and defined the disease-associated haplotype in the 13.6cM interval in 12p11.2-q13.1. This haplotype was shared by all the patients and by some unaffected carriers, suggesting that disease penetration in this family is incomplete. This low penetrance suggests that environmental or other genetic factors modify expression of the disease. Nonparametric two-point and multipoint linkage analyses, which are penetrance-independent, yielded Z(max) LOD scores of 14.2 and 24.9 at D12S345, respectively, strongly supporting the mapping of the parkinsonism locus in this family to 12p11.23-q13.11. This chromosome region is different from any known locus for hereditary parkinsonism, in keeping with the unique genetic features of the parkinsonism in this family. The nomenclature of PARK8 was assigned to the new locus. PMID- 11891825 TI - Protein surveillance machinery in brains with spinocerebellar ataxia type 3: redistribution and differential recruitment of 26S proteasome subunits and chaperones to neuronal intranuclear inclusions. AB - Intracellular aggregates commonly forming neuronal intranuclear inclusions are neuropathological hallmarks of spinocerebellar ataxia type 3 and of other disorders characterized by expanded polyglutamine-(poly-Q) tracts. To characterize cellular responses to these aggregates, we performed an immunohistochemical analysis of neuronal intranuclear inclusions in pontine neurons of patients affected by spinocerebellar ataxia type 3, using a panel of antibodies directed against chaperones and proteasome subunits. A subset of the neuronal intranuclear inclusions stained positively for the chaperones Hsp90alpha and HDJ-2, a member of the Hsp40 family. Most neuronal intranuclear inclusions were ubiquitin positive, suggesting degradation by ubiquitin-dependent proteasome pathways. Surprisingly, only a fraction of neuronal intranuclear inclusions were immunopositive for antibodies directed against subunits of the 20S proteolytic core, whereas most inclusions were stained by antibodies directed against subunits of the 11S and 19S regulatory particles. These results suggest that the proteosomal proteolytic machinery that actively degrades neuronal intranuclear inclusions is assembled in only a fraction of pontine neurons in end stage spinocerebellar ataxia type 3. The dissociation between regulatory subunits and the proteolytic core and the changes in subcellular subunit distribution suggest perturbations of the proteosomal machinery in spinocerebellar ataxia type 3 brains. PMID- 11891826 TI - Destruction of neurons by cytotoxic T cells: a new pathogenic mechanism in Rasmussen's encephalitis. AB - Rasmussen's encephalitis is a progressive epileptic disorder characterized by unihemispheric lymphocytic infiltrates, microglial nodules, and neuronal loss leading to the destruction of the affected hemisphere. In this study, immunohistochemical evaluation of specimens from 11 patients revealed lymphocytic infiltrates that consisted mainly of CD3(+)CD8(+) T cells. Of these cells, 7.0% lay in direct apposition to MHC class I(+) neurons. Confocal laser microscopy revealed that these lymphocytes contained granzyme B in a polar orientation toward these perikarya. Single neurons underwent apoptosis. These findings indicate that a T-cell-mediated cytotoxic reaction induces neuronal death in Rasmussen's encephalitis. This study directly shows, for what we believe is the first time, that a cytotoxic T-cell mechanism contributes to loss of neurons in human brain disease. PMID- 11891827 TI - Role of Caspase-1 in experimental pneumococcal meningitis: Evidence from pharmacologic Caspase inhibition and Caspase-1-deficient mice. AB - Caspase 1 plays a pivotal role in generating mature cytokine interleukin-1beta. Interleukin-1beta is implicated as a mediator of pneumococcal meningitis, both in experimental models and in humans. We demonstrated here that (1) Caspase 1 mRNA and protein expression is upregulated in the brain during experimental pneumococcal meningitis, and (2) Caspase 1 levels are elevated in the cerebrospinal fluid of patients with acute bacterial meningitis. The upregulation/activation of Caspase 1 was associated with increased levels of interleukin-1beta. Depletion of the Caspase 1 gene and pharmacologic blockade of Caspase 1 significantly attenuated the meningitis-induced increase in interleukin 1beta. This was paralleled by a significantly diminished inflammatory host response to pneumococci. The antiinflammatory effect of Caspase 1 depletion or blockade was associated with a marked reduction of meningitis-induced intracranial complications, thus leading to an improved clinical status. In humans, cerebrospinal fluid Caspase 1 levels correlated with the clinical outcome. Thus, pharmacologic inhibition may provide an efficient adjuvant therapeutic strategy in this disease. PMID- 11891828 TI - Adaptive functional changes in the cerebral cortex of patients with nondisabling multiple sclerosis correlate with the extent of brain structural damage. AB - In multiple sclerosis, the mechanisms underlying the accumulation of disability are poorly understood. Recently, it has been suggested that adaptive cortical changes may limit the clinical impact of multiple sclerosis injury. In this study, functional magnetic resonance imaging and a general search method were used to assess patterns of brain activation associated with a simple motor task in 14 right-handed, nondisabled relapsing-remitting multiple sclerosis patients that were compared to those from 15 right-handed, sex- and age-matched healthy volunteers. Also investigated were the extent to which the functional magnetic resonance imaging changes correlated with T2 lesion volume and severity of multiple sclerosis pathology in lesions and normal-appearing brain tissue, measured using magnetisation transfer and diffusion tensor magnetic resonance imaging. Compared to controls, multiple sclerosis patients showed increased activation in the contralateral primary sensorimotor cortex, bilaterally in the supplementary motor area, bilaterally in the cingulate motor area, in the contralateral ascending bank of the sylvian fissure, and in the contralateral intraparietal sulcus. T2 lesion volume was correlated with relative activation in the ipsilateral supplementary motor area, and in the ipsilateral and contralateral cingulate motor area. Average lesion magnetisaiton transfer ratio and average lesion water diffusivity were correlated with relative activation in the contralateral sensorimotor cortex. Average lesion magnetisation transfer ratio was also correlated with relative activation in the ipsilateral cingulate motor area. Average water diffusivity and peak height of the normal-appearing brain tissue diffusivity histogram were both correlated with relative activation in the contralateral intraparietal sulcus. This study shows that cortical activation occurs over a rather distributed sensorimotor network in nondisabled relapsing-remitting multiple sclerosis patients. It also suggests that increased recruitment of this cortical network contributes to the limitation of the functional impact of white matter multiple sclerosis injury. PMID- 11891829 TI - X-linked lissencephaly with absent corpus callosum and ambiguous genitalia (XLAG): clinical, magnetic resonance imaging, and neuropathological findings. AB - X-linked lissencephaly with absent corpus callosum and ambiguous genitalia is a newly recognized syndrome responsible for a severe neurological disorder of neonatal onset in boys. Based on the observations of 3 new cases, we confirm the phenotype in affected boys, describe additional MRI findings, report the neuropathological data, and show that carrier females may exhibit neurological and magnetic resonance imaging abnormalities. In affected boys, consistent clinical features of X-linked lissencephaly with absent corpus callosum and ambiguous genitalia are intractable epilepsy of neonatal onset, severe hypotonia, poor responsiveness, genital abnormalities, and early death. On magnetic resonance imaging, a gyration defect consisting of anterior pachygyria and posterior agyria with a moderately thickened brain cortex, dysplastic basal ganglia and complete agenesis of the corpus callosum are consistently found. Neuropathological examination of the brain shows a trilayered cortex containing exclusively pyramidal neurons, a neuronal migration defect, a disorganization of the basal ganglia, and gliotic and spongy white matter. Finally, females related to affected boys may have mental retardation and epilepsy, and they often display agenesis of the corpus callosum. These findings expand the phenotype of X-linked lissencephaly with absent corpus callosum and ambiguous genitalia, may help in the detection of carrier females in affected families, and give arguments for a semidominant X-linked mode of inheritance. PMID- 11891830 TI - Development of a functional magnetic resonance imaging protocol for intraoperative localization of critical temporoparietal language areas. AB - The aim of this study was to evaluate the use of functional magnetic resonance imaging as an alternative to intraoperative electrocortical stimulation mapping for the localization of critical language areas in the temporoparietal region. We investigated several requirements that functional magnetic resonance imaging must fulfill for clinical implementation: high predictive power for the presence as well as the absence of critical language function in regions of the brain, user independent statistical methodology, and high spatial accuracy. Thirteen patients with temporal lobe epilepsy performed four different functional magnetic resonance imaging language tasks (ie, verb generation, picture naming, verbal fluency, and sentence comprehension) before epilepsy surgery that included intraoperative electrocortical stimulation mapping. To assess the optimal statistical threshold for functional magnetic resonance imaging, images were analyzed with three different statistical thresholds. Functional magnetic resonance imaging information was read into a surgical guidance system for identification of cortical areas of interest. Intraoperative electrocortical stimulation mapping was recorded by video camera, and stimulation sites were digitized. Next, a computer algorithm indicated whether significant functional magnetic resonance imaging activation was present or absent within the immediate vicinity (<6.4mm) of intraoperative electrocortical stimulation mapping sites. In 2 patients, intraoperative electrocortical stimulation mapping failed during surgery. Intraoperative electrocortical stimulation mapping detected critical language areas in 8 of the remaining 11 patients. Correspondence between functional magnetic resonance imaging and intraoperative electrocortical stimulation mapping depended heavily on statistical threshold and varied between patients and tasks. In 7 of 8 patients, sensitivity of functional magnetic resonance imaging was 100% with a combination of 3 functional magnetic resonance imaging tasks (ie, functional magnetic resonance imaging correctly detected all critical language areas with high spatial accuracy). In 1 patient, sensitivity was 38%; in this patient, functional magnetic resonance imaging was included in a larger area found with intraoperative electrocortical stimulation mapping. Overall, specificity was 61%. Functional magnetic resonance imaging reliably predicted the absence of critical language areas within the region exposed during surgery, indicating that such areas can be safely resected without the need for intraoperative electrocortical stimulation mapping. The presence of functional magnetic resonance imaging activity at noncritical language sites limited the predictive value of functional magnetic resonance imaging for the presence of critical language areas to 51%. Although this precludes current replacement of intraoperative electrocortical stimulation mapping, functional magnetic resonance imaging can at present be used to speed up intraoperative electrocortical stimulation mapping procedures and to guide the extent of the craniotomy. PMID- 11891831 TI - Superior oblique myokymia: magnetic resonance imaging support for the neurovascular compression hypothesis. AB - Superior oblique myokymia is a rare movement disorder thought to be caused by vascular compression of the trochlear nerve. Direct display of such neurovascular compression by magnetic resonance imaging has been lacking. The goal of this study was to assess the presence of neurovascular contacts in patients with superior oblique myokymia, using a specific magnetic resonance imaging protocol. A total of 6 patients suffering from right superior oblique myokymia underwent detailed neuro-ophthalmological examination, which showed tonic or phasic eye movement. All patients underwent magnetic resonance imaging, using a magnetic resonance imaging Fourier transform constructive interference in steady-state sequence in combination with magnetic resonance imaging time of flight magnetic resonance arteriography both before and after the administration of Gd-DTPA. With this protocol, the trochlear nerve could be visualized on 11 of 12 sides (92%). Arterial contact was detected at the root exit zone of the symptomatic right trochlear nerve in all 6 patients (100%). No arterial contact was identified at the root exit zone of the asymptomatic left trochlear nerve in any of the 5 left nerves visualized. In conclusion, superior oblique myokymia can result from neurovascular contact at the root exit zone of trochlear nerve, and therefore should be considered among the neurovascular compression syndromes. PMID- 11891832 TI - Comparison of weakness progression in inclusion body myositis during treatment with methotrexate or placebo. AB - We investigated whether 5 to 20mg per week oral methotrexate could slow down disease progression in 44 patients with inclusion body myositis in a randomized double-blind placebo-controlled study over 48 weeks. Mean change of quantitative muscle strength testing sum scores was the primary study outcome measure. Quantitative muscle strength testing sum scores declined in both treatment groups, -0.2% for methotrexate and -3.4% for placebo (95% confidence interval = 2.5% to +9.1% for difference). There were also no differences in manual muscle testing sum scores, activity scale scores and patients' own assessments after 48 weeks of treatment. Serum creatine kinase activity decreased significantly in the methotrexate group. We conclude that oral methotrexate did not slow down progression of muscle weakness but decreased serum creatine kinase activity. PMID- 11891833 TI - A novel tau mutation, S320F, causes a tauopathy with inclusions similar to those in Pick's disease. AB - Mutations in the tau gene cause familial frontotemporal dementia and parkinsonism linked to chromosome 17. In this article, we describe a novel missense mutation, S320F, in the tau gene in a family with presenile dementia. To our knowledge, it is the first mutation to be described in exon 11 of tau. The proband died at age 53 years, after a disease duration of 15 years, and autopsy revealed a neuropathological picture similar to Pick's disease. Recombinant tau protein with the S320F mutation showed a greatly reduced ability to promote microtubule assembly. PMID- 11891834 TI - Brain white matter anatomy of tumor patients evaluated with diffusion tensor imaging. AB - We applied multislice, whole-brain diffusion tensor imaging (DTI) to two patients with anaplastic astrocytoma. Data were analyzed using DTI-based, color-coded images and a 3-D tract reconstruction technique for the study of altered white matter anatomy. Each tumor was near two major white matter tracts, namely, the superior longitudinal fasciculus and the corona radiata. Those tracts were identified using the color-coded maps, and spatial relationships with the tumors were characterized. In one patient the tumor displaced adjacent white matter tracts, whereas in the other it infiltrated the superior longitudinal fasciclus without displacement of white matter. DTI provides new information regarding the detailed relationship between tumor growth and nearby white matter tracts, which may be useful for preoperative planning. PMID- 11891835 TI - Loss of interhemispheric inhibition on the ipsilateral primary sensorimotor cortex in patients with brachial plexus injury: fMRI study. AB - This functional magnetic resonance imaging study verified an antagonistic pattern in which a concomitant deactivation of ipsilateral primary sensorimotor (SM1) was coupled to the contralateral SM1 activation in healthy controls during unimanual hand grasping. Of note, dramatic reduction of ipsilateral SM1 deactivation (loss of antagonistic pattern) was observed during movement of intact hands by patients with unilateral brachial plexus injury. We propose that the disappearance of the antagonistic pattern of SM1 activities in the patients with brachial plexus injury reflects a reduction of interhemispheric inhibition, which may mirror an adaptive mechanism to functional status. PMID- 11891836 TI - Hereditary motor and sensory neuropathy with minifascicle formation in a patient with 46XY pure gonadal dysgenesis: a new clinical entity. AB - This case report is of a patient with 46XY pure gonadal dysgenesis, who presented with chronic progressive motor and sensory polyneuropathy. The sural nerve biopsy exhibited minifascicle formations accompanied by a marked decrease in myelinated fibers. This is the first report of polyneuropathy with minifascicle formations in 46XY pure gonadal dysgenesis. Because a similar polyneuropathy was recently reported in a case with 46XY partial gonadal dysgenesis, it is possible that these cases represent a new type of hereditary motor and sensory neuropathy associated with gonadal dysgenesis. PMID- 11891837 TI - Septo-optic dysplasia associated with a new mitochondrial cytochrome b mutation. AB - We report on a 25-year-old patient with isolated mitochondrial complex III deficiency and a new heteroplasmic mutation (T14849C) in the cytochrome b gene. He suffered from septo-optic dysplasia, retinitis pigmentosa, exercise intolerance, hypertrophic cardiomyopathy, and rhabdomyolysis. A HESX1 mutation was excluded as a cause of his septo-optic dysplasia. Low alpha-tocopherol concentrations in his muscles and an elevated urinary leukotriene E(4) excretion indicate increased production of reactive oxygen species. PMID- 11891839 TI - Familial herpes simplex encephalitis. PMID- 11891838 TI - Mouse glioma gene expression profiling identifies novel human glioma-associated genes. AB - Based on previous studies demonstrating increased RAS activity in human astrocytomas, we developed a transgenic mouse model (B8) that targets an activated RAS molecule to astrocytes. Within 3 to 4 months after birth, these mice develop high-grade astrocytomas that are histologically identical to human astrocytomas. To characterize genetic events associated with B8 mouse astrocytoma formation, we employed comparative gene expression profiling of wild-type cultured mouse astrocytes, non-neoplastic B8 astrocytes, B8 astrocytoma cultures, and two other astrocytoma cultures from independently derived RAS transgenic mouse lines. We identified several classes of gene expression changes, including those associated with the non-neoplastic state in the B8 transgenic mouse, those associated with astrocytoma formation, and those specifically associated with only one of the three independently derived transgenic mouse astrocytomas. Differential expression of several unique genes was confirmed at the protein level in both the RAS transgenic mouse astrocytomas and two human glioblastoma multiforme cell lines. Furthermore, reexpression of one of these downregulated astrocytoma-associated proteins, GAP43, resulted in C6 glioma cell growth suppression. The use of this transgenic mouse model to identify novel genetic changes that might underlie the pathogenesis of human high-grade astrocytomas provides a unique opportunity to discover future targets for brain tumor therapy. PMID- 11891840 TI - Effective treatment for essential tremor. PMID- 11891842 TI - Spinocerebellar ataxia type 10 in the French population. PMID- 11891845 TI - DNA replication and nuclear architecture. AB - The model of in situ DNA replication provided by immunofluorescence and confocal imaging is compared with observations obtained by electron microscopic studies. Discrepancies between both types of observations call into question the replication focus as a persistent nuclear structure and as a replication entity where DNA replication takes place. Most electron microscopic analyses reveal that replication sites are confined to dispersed chromatin areas at the periphery of condensed chromatin, and the distribution of replication factors exhibits the same localization pattern. Moreover, rapid migration of newly synthesized DNA from the replication sites towards the interior of condensed chromatin regions obviously takes place during S-phase. It implies modifications of replication domains, hardly detectable by fluorescence microscopy. The confrontation of different observations carried out at light microscopic or electron microscopic levels of resolution lead to a conclusion that a combination of in vivo fluorescence analysis with a subsequent ultrastructural investigation performed on the same cells will represent an optimal approach in future studies of nuclear functions in situ. PMID- 11891846 TI - Hormonal regulation of the c-fms proto-oncogene in breast cancer cells is mediated by a composite glucocorticoid response element. AB - We have previously reported that glucocorticoids markedly increase and anti glucocorticoids (such as RU-486) block c-fms RNA and protein expression in some breast cancer cell lines, but not in others, and that this increase is the consequence of increased transcription from the first, epithelial cell-specific promoter of the c-fms gene (encoding CSF-1R, macrophage colony-stimulating factor receptor). Employing DNaseI protection and electrophoretic mobility shift assays (EMSA), we now demonstrate that DNA-transcription factor protein complexes are formed on the c-fms first promoter at a composite regulatory element containing overlapping binding sites for AP-1 proteins, bHLH factors, and the glucocorticoid receptor (GR). Competition studies indicate that transcription factor proteins bind the AP-1 site and the GR element (GRE) and both GR and AP-1 proteins are involved in DNA-protein complex formation. The complexes differ in quantity and glucocorticoid inducibility in the different breast cancer cell lines studied depending on whether the promoter responds to glucocorticoid stimulation. Transient transfection of promoter/reporter gene constructs resulted in reduced basal transcription activity of this promoter and lack of glucocorticoid stimulation when the AP-1 site was mutated. We conclude that AP-1 proteins, GR and associated co-factors regulate transcription from the c-fms first promoter and that differences in recruitment of the various components are responsible for cell specific repression and activation of this gene in breast carcinoma cell lines. PMID- 11891847 TI - Nonenzymatic glycation of histones in vitro and in vivo. AB - Purified histones in solution, purified nuclei, or whole endothelial cells in cell culture were used to study the reactivity of histones with various sugars. The sugar incubation of purified histones produced nonenzymatic glycation and formation of histone cross-links showing disappearance of individual histone molecules and appearance of dimers and polymers in SDS-PAGE. In solution, core histones react considerably faster with sugars as compared to H1 histones. In sugar-incubated nuclei where histones are nucleosomally organized, H1 histones, which are located at the periphery of the nucleosome, and H2A-H2B dimers, which are associated with the central H3(2)-H4(2) tetramer, are more reactive as compared to H3 and H4 histones, which are most protected from the glycation reaction. Our in vivo experiments using endothelial cells show that high concentrations of ribose are able to generate protein cross-links paralleled by apoptotic cell death. High concentrations of glucose or fructose do not increase histone glycation or cell death, even after 60 days of incubation of endothelial cells. In long-time glucose- or fructose-treated cells, under nondenaturing and nonreducing SDS-PAGE conditions part of the H3 histones shifted away from their normal location. Because it is known that the mitochondrial production of reactive oxygen species (ROS) increases after hyperglycaemia, we hypothesize that ROS could be responsible for the formation of a disulphide bridge between the side chain of the cysteine residues of H3 molecules. PMID- 11891848 TI - Involvement of cAMP but not PKA in the increase of corticosterone secretion in rat zona fasciculata-reticularis cells by aging. AB - The effects and mechanisms of aging on corticosterone secretion in zona fasciculata-reticularis (ZFR) cells of ovariectomized (Ovx) rats were studied. Young (3-month) and old (24-month) female rats were Ovx for 4 days before decapitation. ZFR cells were isolated and incubated with different hormones or reagents at 37 degrees C for 30 min. Aging increased the basal secretion of corticosterone both in vivo and in vitro. The adrenocorticotropin (ACTH)-, forskolin-, 3-isobutyl-l-methylxanthine (IBMX)-, 8-bromo-adenosine 3',5'-cyclic monophosphate (8-Br-cAMP)-, and ovine prolactin (oPRL)-stimulated release of corticosterone by ZFR cells was greater in old than in young Ovx rats. H89, an inhibitor of protein kinase A (PKA), decreased the production of corticosterone in ZFR cells from young but not old Ovx rats. Forskolin-, or IBMX-induced production of cAMP was greater in old than in young Ovx animals, which correlated with the increase of corticosterone production by aging. The activity of 11 beta hydroxylase that converts deoxycorticosterone (DOC, 10(-9) or 10(-8) M) to corticosterone in rat ZFR cells was decreased by age. However, the corticosterone production in response to high dose of DOC (10(-7) M) was indifferent between young and old groups. These results suggest that aging increases corticosterone production in Ovx rats via a mechanism in part associated with an increase of adenylyl cyclase activity and a decrease of phosphodiesterase activity, and then an increase of the generation of cAMP, but not related to either PKA activity or 11 beta-hydroxylase. PMID- 11891849 TI - Phosphorylation of tyrosine 256 facilitates nuclear import of atypical protein kinase C. AB - Herein, we employed a combined approach of molecular modeling and site-directed mutagenesis to address the role of tyrosine phosphorylation in transport of atypical protein kinase C (aPKC) into the nucleus. Computer modeling of the three dimensional structure of the aPKC catalytic core, reveals that tyrosine 256 (Tyr256) is located at the lip of the activation loop and is conserved among members of the aPKC family, iota/lambda and zeta. Based on these findings, we examined whether tyrosine phosphorylation of aPKC on the activation lip may facilitate nuclear import. An antiserum was generated that selectively recognizes the phosphorylated Tyr256 residue in aPKC. By isolating nuclei of PC12 cells and immunoprecipitating aPKC with Ab-PY256, we observed that Tyr256 is rapidly phosphorylated upon NGF treatment prior to entry of aPKC into the nucleus. aPKC was observed to exclusively bind to importin-beta. The interaction between importin-beta and aPKC was enhanced upon tyrosine phosphorylation of aPKC and binding was abrogated when Tyr256 was mutated to phenylalanine. We propose that phosphorylation of aPKC at Tyr256 induces a conformation, whereby, the arginine rich NLS is exposed, which then binds importin-beta leading to import of aPKC into the nucleus. Altogether, these findings document a novel role for the tyrosine phosphorylation in regulating import of atypical PKC into the nucleus. PMID- 11891850 TI - Suppression of tissue factor expression, cofactor activity, and metastatic potential of murine melanoma cells by the N-terminal domain of adenovirus E1A 12S protein. AB - Tissue factor, the cellular initiator of blood coagulation, has been implicated as a determinant of metastatic potential in human melanoma cells. Here, we report that differential expression of tissue factor in murine melanoma cell lines of known metastatic behavior is mediated by AP-1-dependent and 12S E1A oncoprotein repressible gene transcription. When compared to weakly metastatic C10 cells, highly metastatic M4 cells possessed elevated levels of tissue factor cofactor activity, transfected promoter activity, and heterodimeric AP-1 DNA-binding complexes containing Fra-1. Transient co-expression of the adenovirus E1A 12S oncoprotein strongly repressed transcription of an AP-1-driven tissue factor reporter gene indicating the additional requirement of N-terminal E1A-interacting coactivators. Stable expression of E1A mutants defective in CBP/p300-binding failed to suppress tissue factor expression and experimental metastasis by M4 cells while clones expressing wild type E1A exhibited greatly reduced tissue factor cofactor activity and metastatic potential in vivo. Overexpression of functional tissue factor in cells containing wild type E1A failed to restore the highly metastatic M4 phenotype suggesting that additional E1A-responsive and CBP/p300-dependent genes are required to facilitate metastasis of murine melanoma cells demonstrating high tissue factor expression and cofactor activity. PMID- 11891851 TI - Orphan nuclear receptor binding site in the human inducible nitric oxide synthase promoter mediates responsiveness to steroid and xenobiotic ligands. AB - Constitutive androstane receptor (CAR) and pregnane X receptor (PXR) are members of the nuclear receptor superfamily that regulate target gene transcription in a ligand-dependent manner. CAR and PXR have a rather broad, overlapping set of ligands that range from natural steroids to xenobiotics and also recognize similar DNA binding sites, referred to as response elements (REs), primarily in promoter regions of cytochrome P450 (CYP) genes. In this study, a CAR and PXR RE, composed of a direct repeat of two GGTTCA motifs in a distance of 4 nucleotides (DR4), was identified in the promoter of the human inducible nitric oxide (NO) synthase (iNOS) gene, which is the first nuclear receptor binding site reported for this promoter. In a heterologous promoter context, the DR4-type sequence also acts as a functional RE for the nuclear receptors for 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25OH2D3) and 3,5,3'-triiodothyronine (T3), VDR and T3R. However, in a direct competition of CAR, PXR, VDR, and T3R, the PXR-retinoid X receptor (RXR) complex appears to be the dominant regulator on the iNOS DR4-type RE. In the natural iNOS promoter context, the DR4-type RE specifically mediates downregulation of promoter activity by the testosterone metabolite androstanol through CAR-RXR heterodimers and upregulation by the xenobiotic drug clotrimazole through PXR-RXR heterodimers. These results were confirmed on the level of mRNA expression. Since an iNOS-induced production of NO is known to influence inflammation and apoptosis, a CAR- and PXR-regulated iNOS activity may explain a modulatory effect of steroids and xenobiotics on these cellular processes. PMID- 11891852 TI - Nigericin inhibits insulin-stimulated glucose transport in 3T3-L1 adipocytes. AB - We used nigericin, a K+/H+ exchanger, to test whether glucose transport in 3T3-L1 adipocytes was modulated by changes in intracellular pH. Our results showed that nigericin increased basal but decreased insulin-stimulated glucose uptake in a time- and dose-dependent manner. Whereas the basal translocation of GLUT1 was enhanced, insulin-stimulated GLUT4 translocation was inhibited by nigericin. On the other hand, the total amount of neither transporter protein was altered. The finding that insulin-stimulated phosphoinositide 3-kinase (PI 3-kinase) activity was not affected by nigericin implies that nigericin exerted its inhibition at a step downstream of PI 3-kinase activation. At maximal dose, nigericin rapidly lowered cytosolic pH to 6.7; however, this effect was transient and cytosolic pH was back to normal in 20 min. Removal of nigericin from the incubation medium after 20 min abolished its enhancing effect on basal but had little influence on its inhibition of insulin-stimulated glucose transport. Moreover, lowering cytosolic pH to 6.7 with an exogenously added HCl solution had no effect on glucose transport. Taken together, it appears that nigericin may inhibit insulin stimulated glucose transport mainly by interfering with GLUT4 translocation, probably by a mechanism not related to changes in cytosolic pH. PMID- 11891853 TI - Modulation of the proliferation and differentiation of human mesenchymal stem cells by copper. AB - Copper plays important functional roles in bone metabolism and turnover. It is known that it is essential for normal growth and development of the skeleton in humans and in animals. Although at present the exact role that copper plays in bone metabolism is unknown, bone abnormalities are a feature of severe copper deficiency. Osteoblasts are derived from mesenchymal stem cells (MSCs) present in bone marrow stroma, which are able to differentiate into bone, adipocytes, and other cell phenotypes. Excess adipogenesis in postmenopausal women may occur at the expense of osteogenesis and, therefore, may be an important factor in the fragility of postmenopausal bone. The purpose of this study was to evaluate whether an increase of the extracellular concentration of copper affects the ability of MSCs to differentiate into osteoblasts or adipocytes. The results showed that copper modified both the differentiation and the proliferative activity of MSCs obtained from postmenopausal women. Copper (50 microM) diminished the proliferation rate of MSCs, increasing their ability to differentiate into the osteogenic and the adipogenic lineages. Copper induced a 2 fold increase in osteogenic differentiation of MSCs, measured as a increase in calcium deposition. Copper (5 and 50 microM) diminished the expression of alkaline phosphatase (50 and 80%, respectively), but induced a shift in the expression of this enzyme to earlier times during culture. Copper also induced a 1.3-fold increase in the adipogenic differentiation of MSCs. It is concluded that copper stimulates MSC differentiation, and that this is preferentially towards the osteogenic lineage. PMID- 11891854 TI - Analysis of the carboxypeptidase D cytoplasmic domain: Implications in intracellular trafficking. AB - Metallocarboxypeptidase D (CPD) is a type 1 transmembrane protein that functions in the processing of proteins that transit the secretory pathway. Previously, CPD was found to be enriched in the trans Golgi network (TGN) and to cycle between this compartment and the cell surface. In the present study, the roles of specific regions of the CPD cytosolic tail in intracellular trafficking were investigated in the AtT-20 cell line. When the CPD transmembrane region and cytosolic tail are attached to the C-terminus of albumin, this protein is retained in the TGN and cycles to the cell surface. Deletion analysis indicates that a C-terminal region functions in TGN-retention; removal of 10 amino acids from the C-terminus greatly increases the amount of fusion protein that enters nascent vesicles, which bud from the Golgi, but does not affect the half-life of the fusion protein or the ability of cell surface protein to return to the TGN. Because the 10-residue deletion disrupts a casein kinase 2 (CK2) consensus site, the two Thr in this site (TDT) were mutated to either Ala (ADA) or Glu (EDE). Neither mutation has an increased rate of budding from the TGN, although the ADA mutant has a shorter half-life than either the wild type sequence or the EDE mutant. Adaptor protein-1 and -2 bind to most of the deletion mutants, the EDE point mutant, and the CK2-phosphorylated CPD tail, but not to the wild type tail. Taken together, these results suggest that CPD localization to the TGN requires both static retention involving the C-terminal domain and phosphorylation at a CK2 site, which regulates the binding of adaptor proteins. PMID- 11891855 TI - Reduced CpG methylation is associated with transcriptional activation of the bone specific rat osteocalcin gene in osteoblasts. AB - Chromatin remodeling of the bone-specific rat osteocalcin (OC) gene accompanies the onset and increase in OC expression during osteoblast differentiation. In osseous cells expressing OC, the promoter region contains two nuclease hypersensitive sites that encompass the elements that regulate basal tissue specific and vitamin D-enhanced OC transcription. Multiple lines of evidence indicate that DNA methylation is involved in maintaining a stable and condensed chromatin organization that represses eukaryotic transcription. Here we report that DNA methylation at the OC gene locus is associated with the condensed chromatin structure found in cells not expressing OC. In addition, we find that reduced CpG methylation of the OC gene accompanies active transcription in ROS 17/2.8 rat osteosarcoma cells. Interestingly, during differentiation of primary diploid rat osteoblasts in culture, as the OC gene becomes increasingly expressed, CpG methylation of the OC promoter is significantly reduced. Inhibition of OC transcription does not occur by a direct mechanism because in vitro methylated OC promoter DNA is still recognized by the key regulators Runx/Cbfa and the vitamin D receptor complex. Furthermore, CpG methylation affects neither basal nor vitamin D-enhanced OC promoter activity in transient expression experiments. Together, our results indicate that DNA methylation may contribute indirectly to OC transcriptional control in osteoblasts by maintaining a highly condensed and repressed chromatin structure. PMID- 11891856 TI - Cytokine regulatory effects on alpha-1 proteinase inhibitor expression in NOD mouse islet endothelial cells. AB - Human microvascular islet endothelial cells (IEC) exhibit specific morphological and functional characteristics that differ from endothelia derived from other organs. One of these characteristics is the expression of alpha-1 proteinase inhibitor (Api). In this study, we observed its expression in nonobese diabetic (NOD) mouse IEC, in relation to the occurrence of type 1 diabetes and in response to cytokines, namely IL-1 beta and IL-10. In addition, IL-10-deficient NOD mice as well as IL-10 transgenic NODs were studied. Results have demonstrated that Api expression is: (i) highly specific for IEC in NOD mouse islets, as for humans; (ii) linked to the occurrence of early type 1 diabetes, and iii) strongly modulated by Th1 and Th2 cytokines. In fact, Api mRNA found in pre-diabetic NOD animals is significantly reduced when they become hyperglycemic and disappears by 25 weeks of age, when mice are diabetic. Moreover, Api mRNAs are never seen in nondiabetic controls. Furthermore, in cultured NOD IEC, Api expression is downregulated by the addition of IL-1 beta and is upregulated by IL-10; it is always absent in IL-10-deficient NOD mice and overexpressed in IL-10 transgenic NODs, thus further supporting that this cytokine upregulates Api expression. PMID- 11891857 TI - Overexpression of murine phosphatidylinositol 4-phosphate 5-kinase type Ibeta disrupts a phosphatidylinositol 4,5 bisphosphate regulated endosomal pathway. AB - The type I phosphatidylinositol 4-phosphate 5-kinases (PI4P5K) phosphorylate phosphatidylinositol 4-phosphate [PI(4)P] to produce phosphatidylinositol 4,5 bisphosphate [PI(4,5)P2]. PI(4,5)P2 has been implicated in signal transduction, receptor mediated endocytosis, vesicle trafficking, cytoskeletal structure, and membrane ruffling. However, the specific type I enzymes associated with the production of PI(4,5)P2 for the specific cellular processes have not been rigorously defined. Murine PI4P5K type Ibeta (mPIP5K-Ibeta) was implicated in receptor mediated endocytosis through the isolation of a truncated and inactive form of the enzyme that blocked the ligand-dependent downregulation of the colony stimulating factor-1 receptor. The present study shows that enforced expression of mPIP5K-Ibeta in 293T cells resulted in the accumulation of large vesicles that were linked to an endosomal pathway. Similar results were obtained after the expression of the PI(4,5)P2-binding pleckstrin homology (PH) domain of phospholipase-Cdelta (PLC-delta). Analysis of the conserved domains of mPIP5K Ibeta led to the identification of dimerization domains in the N- and C-terminal regions. Enforced expression of the individual dimerization domains interfered with the proper subcellular localization of mPIP5K-Ibeta and the PLC-delta-PH domain and blocked the accumulation of the endocytic vesicles induced by these proteins. In addition to regulating early steps in endocytosis, these results suggest that mPIP5K-Ibeta acts through PI(4,5)P2 to regulate endosomal trafficking and/or fusion. PMID- 11891858 TI - Interaction of the ADP-ribosylating enzyme from the hyperthermophilic archaeon S. solfataricus with DNA and ss-oligo deoxy ribonucleotides. AB - The DNA-binding ability of the poly-ADPribose polymerase-like enzyme from the extremely thermophilic archaeon Sulfolobus solfataricus was determined in the presence of genomic DNA or single stranded oligodeoxyribonucleotides. The thermozyme protected homologous DNA against thermal denaturation by lowering the amount of melted DNA and increasing melting temperature. The archaeal protein induced structural changes of the nucleic acid by modifying the dichroic spectra towards a shape typical of condensing DNA. However, enzyme activity was slightly increased by DNA. Competition assays demonstrated that the protein interacted also with heterologous DNA. In order to characterize further the DNA binding properties of the archaeal enzyme, various ss-oligodeoxyribonucleotides of different base composition, lengths (12-mer to 24-mer) and structure (linear and circular) were used for fluorescence titration measurements. Intrinsic fluorescence of the archaeal protein due to tryptophan (excitation at 295 nm) was measured in the presence of each oligomer at 60 degrees C. Changes of tryptophan fluorescence were induced by all compounds in the same range of base number per enzyme molecule, but independently from the structural features of oligonucleotides, although the protein exhibited a slight preference for those adenine-rich and circular. The binding affinities were comparable for all oligomers, with intrinsic association constants of the same order of magnitude (K=10(6) M(-1)) in 0.01 M Na-phosphate buffer, pH 8.0, and accounted for a "non specific" binding protein. Circular dichroism analysis showed that at 60 degrees C the native protein was better organized in a secondary structure than at 20 degrees C. Upon addition of oligonucleotides, enzyme structure was further stabilized and changed towards a beta-conformation. This effect was more marked with the circular oligomer. The analysed oligodeoxyribonucleotides slightly enhanced enzyme activity with the maximal increase of 50% as compared to the control. No activation was observed with the circular oligomer. PMID- 11891859 TI - High stability binding of poly(ADPribose) polymerase-like thermozyme from S. solfataricus with circular DNA. AB - The poly(ADPribose) polymerase-like thermozyme from the hyperthermophilic archaeon S. solfataricus was found to bind DNA with high affinity and non specifically. Binding was independent of base composition and length of the nucleic acid, and the protein showed a slight preference for the circular structure. By using pCMV-Neo-Bam plasmid as experimental model, the behaviour of the thermozyme upon binding with either circular or linear plasmid was analyzed. pCMV-Neo-Bam has a single HindIII site that allows to obtain the linear structure after digestion with the restriction enzyme. Intrinsic tryptophan-dependent fluorescence of poly(ADPribose) polymerase-like thermozyme noticeably changed upon addition of either circular or linear plasmid, showing the same binding affinity (K=2 x 10(9) M-1). However, experiments of protection against temperature and DNase I gave evidence that the thermozyme formed more stable complexes with the circular structure than with the linear pCMV-Neo-Bam. Increasing temperature at various DNA/protein ratios had a double effect to reduce the amount of circular DNA undergoing denaturation and to split the melting point towards higher temperatures. Nil or irrelevant effect was observed with the linear form. Similarly, DNase acted preferentially on the linear plasmid/protein complexes, producing an extensive digestion even at high protein/DNA ratios, whereas the circular plasmid was protected by the thermozyme in a dose-dependent manner. The complexes formed by archaeal poly(ADPribose) polymerase (PARPss) with the circular plasmid were visualized by bandshift experiments both with ethidium bromide staining and by labelling the circular plasmid with 32P. The stability of complexes was tested as a function of enzyme concentration and in the presence of a cold competitor and of 0.1% SDS. From the performed experiments, a number of 3-10 base pairs bound per molecule of enzyme was calculated, indicating a high frequency of binding. The presence of circular DNA was also able to increase by 80% the poly(ADPribose)polymerase-like activity, as compared to 25% activation induced by the linear pCMV-Neo-Bam. PMID- 11891860 TI - Characterization of the osteoblast-like cell phenotype under microgravity conditions in the NASA-approved Rotating Wall Vessel bioreactor (RWV). AB - Weightlessness induces bone loss in humans and animal models. We employed the NASA-approved Rotating Wall Vessel bioreactor (RWV) to develop osteoblast-like cell cultures under microgravity and evaluate osteoblast phenotype and cell function. Rat osteoblast-like cell line (ROS.SMER#14) was grown in the RWV at a calculated gravity of 0.008g. For comparison, aliquots of cells were grown in conventional tissue culture dishes or in Non-Rotating Wall Vessels (N-RWV) maintained at unit gravity. In RWV, osteoblasts showed high levels of alkaline phosphatase expression and activity, and elevated expression of osteopontin, osteocalcin, and bone morphogenetic protein 4 (BMP-4). In contrast, the expression of osteonectin, bone sialoprotein II and BMP-2 were unaltered compared to cells in conventional culture conditions. These observations are consistent with a marked osteoblast phenotype. However, we observed that in RWV osteoblasts showed reduced proliferation. Furthermore, DNA nucleosome-size fragmentation was revealed both morphologically, by in situ staining with the Thymine-Adenine binding dye bis-benzimide, and electrophoretically, by DNA laddering. Surprisingly, no p53, nor bcl-2/bax, nor caspase 8 pathways were activated by microgravity, therefore the intracellular cascade leading to programmed cell death remains to be elucidated. Finally, consistent with an osteoclast stimulating effect by microgravity, osteoblasts cultured in RWV showed upregulation of interleukin-6 (IL-6) mRNA, and IL-6 proved to be active at stimulating osteoclast formation and resorbing activity in vitro. We conclude that under microgravity, reduced osteoblast life span and enhanced IL-6 expression may result in inefficient osteoblast- and increased osteoclast activity, respectively, thus potentially contributing to bone loss in individuals subjected to weightlessness. PMID- 11891861 TI - Soluble fragment of P-cadherin adhesion protein found in human milk. AB - Classical cadherins such as E- and P-cadherin are transmembrane proteins that mediate specific cell-to-cell adhesion and are important to tissue development and function. Cadherin function can be modulated by various means, including proteolytic cleavage of the extracellular adhesion domain from the cells' surface, yielding large soluble fragments termed (soluble) sE- or sP-cadherin. In people with certain carcinomas, sE-cadherin can be detected at elevated levels in the serum and sometimes can serve as a prognostic marker. Soluble E-cadherin also is found in urine of patients with bladder cancer. In addition to being present in bodily fluids of cancer patients, sE- and sP-cadherin are present in the serum of healthy people, suggesting that shedding of cadherins is a normal event. Here, we report high levels of 80 kDa sP-cadherin in human milk. In the lactating mammary gland tissue, P-cadherin appears to be a protein secreted by epithelial cells, rather than an adhesion protein. This is in contrast to the non-lactating mammary gland where P-cadherin is restricted to myoepithelial cells, and is present at sites of cell-cell contact. PMID- 11891862 TI - Osteocytes inhibit osteoclastic bone resorption through transforming growth factor-beta: enhancement by estrogen. AB - Osteocytes are the most abundant cells in bone and distributed throughout the bone matrix. They are connected to the each other and to the cells on the bone surface. Thus, they may also secrete some regulatory factors controlling bone remodeling. Using a newly established osteocyte-like cell line MLO-Y4, we have studied the interactions between osteocytes and osteoclasts. We collected the conditioned medium (CM) from MLO-Y4 cells, and added it into the rat osteoclast cultures. The conditioned medium had no effect on osteoclast number in 24-h cultures, but it dramatically inhibited resorption. With 5, 10, and 20% CM, there was 25, 39, and 42% inhibition of resorption, respectively. Interestingly, the inhibitory effect was even more pronounced, when MLO-Y4 cells were pretreated with 10(-8) M 17-beta-estradiol. With 5, 10, and 20% CM, there was 46, 51, and 58% of inhibition. When the conditioned medium was treated with neutralizing antibody against transforming growth factor-beta (TGF-beta), the inhibitory effect was abolished. This suggests that osteocytes secrete significant amounts of TGF-beta, which inhibits bone resorption and is modulated by estrogen. RT-PCR and Western blot analysis show that in MLO-Y4 cells, the prevalent TGF-beta isoform is TGF-beta3. We conclude that osteocytes have an active, inhibitory role in the regulation of bone resorption. Our results further suggest a novel role for TGF-beta in the regulation of communication between different bone cells and suggest that at least part of the antiresorptive effect of estrogen in bone could be mediated via osteocytes. PMID- 11891863 TI - Cardiac troponin I sense-antisense RNA duplexes in the myocardium. AB - Natural antisense RNA is now thought to regulate, at least in part, a growing number of eukaryotic genes. It is becoming increasingly apparent that such endogenous antisense RNA molecules may modulate gene expression in a manner analogous to synthetic oligomers. Here, we report the detection of antisense orientated RNA transcripts of cardiac specific troponin I in rat and human myocardium. Interestingly, the different sizes of the rat and human antisense cTNI transcripts suggest species-specific reverse transcription initiation sites. Moreover, for the first time in cardiomyocytes, we could demonstrate in vivo duplex formation between sense and antisense transcripts. The existence of antisense-sense duplexes represents compelling evidence and a potential mechanism for endogenous antisense transcript-mediated modulation of mRNA translation. The potential effect of attenuating translation was illustrated by in vitro and in vivo model systems. Testing several oligonucleotides based on the natural antisense sequences, the optimal region for inhibition of translation was identified as being close to the translational start codon. PMID- 11891865 TI - Immunohistochemical localization of type 2 inositol 1,4,5-trisphosphate receptor to the nucleus of different mammalian cells. AB - The inositol 1,4,5-trisphosphate receptor (InsP3R) is a ligand-gated Ca2+ channel responsible for the release of Ca2+ from intracellular stores in the response of a wide variety of cells to external stimuli. Molecular cloning studies have revealed the existence of three types of InsP3R encoded by distinct genes. In the study presented here, we used selective anti-InsP3R antibodies to determine the intracellular location of each InsP3R subtype in bovine aortic endothelial cells, bovine adrenal glomerulosa cells, and COS-7 cells. InsP3R1 was found to be widely distributed throughout the cytosol and most abundantly in the perinuclear region identified as the endoplasmic reticulum (co-localization with protein disulfide isomerase). The intracellular location of InsP3R3 was similar to that of InsP3R1. Surprisingly, InsP3R2 was found mostly associated to the cell nucleus. This observation was made with two antibodies recognizing different epitopes on InsP3R2. Binding studies revealed the presence of a high affinity-binding site for [3H] InsP3 on purified nuclei from bovine adrenal cortex. Confocal images showed that InsP3R2 was not confined to the nuclear envelope but was distributed relatively uniformly within the nucleus. Our results demonstrate that the three types of InsP3R are not similarly distributed within a specific cell type. Our results also suggest the existence of an intranuclear membrane network on which InsP3R2 is abundantly expressed. PMID- 11891864 TI - O-glycosylation of the nuclear forms of Pax-6 products in quail neuroretina cells. AB - Many transcription factors are demonstrated as being glycosylated with O-N acetylglucosamine (GlcNAc) residue in transfected insect cell lines, but rarely in the original cells. For the first time, we demonstrate the O-GlcNAc modification of the p48/p46 Pax-6 gene (a developmental control gene involved in the eye morphogenesis) products in the quail neuroretina (QNR). In conjunction with a systematic PNGase F treatment, we used wheat germ agglutinin (WGA) binding, in vitro labeling with bovine galactosyltransferase, and labeling of cultured QNR with [14C]GlcNH2. Glycosylated forms of Pax-6 proteins were found in the nucleus of the neuroretina cells. WGA-selected Pax-6 proteins produced in the reticulocyte lysate were able to bind a DNA target, as well as to the unglycosylated form. The O-GlcNAc may, however, modulate protein interactions, mainly with other factors involved in the transcription process. Characterization of products released after reductive alkaline treatment of the proteins clearly demonstrates that N-acetylglucosamine is directly linked to serine or threonine residues. Examination of Pax-6 primary sequence allowed us to determine potential O-GlcNAc attachment sites. Most of these expected glycosylation sites appear to be located on the two DNA binding domains and on the carboxyterminal transactivation domain, while experimental evidence taken from WGA-selected proteins experiment points in favor of a main localization on the paired-box domain. PMID- 11891866 TI - Interrelated fellowships: the optimum training experience? PMID- 11891867 TI - Dr. Zhong Wei Chen: a microsurgery pioneer. PMID- 11891868 TI - A reconstructed digit by transplantation of a second toe for control of an electromechanical prosthetic hand. AB - The treatment options for the loss of an entire human hand and part of the forearm are currently limited to the transplantation of toe(s) to the amputation stump or a Krukenberg's bifurcation hand, and using a cosmetic or functional prosthesis. The functional prosthetic hand, such as the prevailing myoelectrically controlled prosthetic hand, has an action accuracy that is affected by many factors. The acceptance rate of the three planes freedom myoelectronic hand by the patients was 46-90% because of poor function caused by the weakness of signal and strong external interference. In this report, the left second toe was transplanted to the patient's forearm amputation stump. Mandates from the brain are relayed by the action of this reconstructed digit, to control a special designed multidimension freedom electronic prosthetic hand. After rehabilitation and adaptation training, the correct recognition rate of the electronic prosthetic hand controlled by this reconstructed digit is a remarkable 100%. PMID- 11891869 TI - Electrophysiologic findings and muscle strength grading in brachioplexopathies. AB - The electrophysiological evaluations and the British Medical Research Council (MRC) scale (0-5) findings of target muscles in brachioplexopathies before surgery and 1 year postsurgery were conducted. Each component of the brachial plexus was analyzed in 15 patients with injuries, among them, to 5 roots, 19 trunks, 7 cords, and 13 terminal nerves. In each of these cases, neurolysis and/or nerve transfer and/or neurotization were performed, within 3 weeks to 6 months after the injury was incurred, to ameliorate the resulting severe disabilities. The degrees of impairment were graded using a modified version of Dumitru's and Wilbourn's scale (mild: normal to slight decrease of SNAP amplitude and CMAP amplitude, and occasional denervation; moderate: profound decrease of SNAP amplitude and CMAP amplitude, constant denervation, and normal to slight decrease in motor unit recruitment; severe: absent SNAP amplitude, absent CMAP amplitude, marked denervation, and profound decrease or no volitional motor unit recruitment. mild = 1; moderate = 2; severe = 3). The motor power of the target muscles was graded through MRC scores. The presurgical versus postsurgical differences in the severity of the injury to each brachial plexus component, and differences in the grading of target muscle power, were calculated through the Wilcoxon signed-rank test. The presurgical degrees of the severity of injury, as measured by the electromyography (EMG) were 3.00 +/- 0.00 (mean +/- SD) in root, 2.84 +/- 0.50 in trunk, 3.00 +/- 0.00 in cord, and 2.85 +/- 0.38 in terminal nerves. The postsurgical results were 2.60 +/- 0.55 in root, 2.53 +/- 0.70 in trunk, 2.43 +/- 0.53 in cord, and 1.77 +/- 0.73 in terminal nerves. There was significant improvement at the trunk, cord, and terminal nerve levels after repair, but not at the root levels. Moreover, although the MRC grading showed significant motor recovery in the infraspinatus, deltoid, biceps, and triceps muscles, there was little apparent improvement in the pectoralis major, EDC, APB, and ADM muscles. Nerve repair was notably successful in all plexuses except at the root level. However, our cases demonstrated only poor motor power gains in the forearm and the hand muscles. Consequently, future surgical techniques for brachioplexopathy repairs need further improvement. PMID- 11891870 TI - Effect of hyperbaric oxygen on pedicle flaps with compromised circulation. AB - The effect of hyperbaric oxygen (HBO) on the survival rate of experimental rat pedicle island flaps with arterial, venous, and combined arteriovenous insufficiency was evaluated. Forty male Wistar rats with pedicle island flaps were divided into four groups with different types of vascular status. Another 40 male Wistar rats, also divided into four groups, were reconstructed in the same manner, but were also exposed to HBO. The results were evaluated using a laser Doppler flowmeter and an estimation of the length of the surviving tissue of the flaps. In evaluations using Kruskal-Wallis test, there was a significant difference in the survival tissue length and mean LD flows among our four untreated groups (P < 0.05). We considered the experimental model defined by Tzusuki and colleagues suitable for our study. Using a Mann-Whitney test, the differences in flap tissue survival length between each type of vascular insufficiency of HBO-treated and untreated groups was significant (P < 0.05). This finding indicates that the survival length was directly improved by the HBO treatment for all type of vascular insufficiency. HBO treatment increased the percentage of survival length and mean LD flows of axial pattern skin flaps with all type of vascular insufficiency. This effect, however, was greatest in the arterial insufficiency flaps. PMID- 11891871 TI - Auxiliary liver transplantation with portal arterialization in the rat: description of a new model. AB - In recent years, portal arterialization has been used in liver transplantation to increase the portal flow, as a solution for singular technical problems. We have developed a new auxiliary liver transplantation model in the rat with portal arterialization, so the native hepatic hilium remains untouched, consisting on a graft with a previous 70% hepatectomy. It is sited on the right renal bed, joining the infrahepatic inferior vena cava (IVC) of the graft with the recipient IVC. With an abdominal aortic graft, we connect the recipient aorta with the portal vein from the auxiliary liver. All the animals survived at the seventh day. No thrombosis was seen in any graft and an important rejection was observed in all the fields. We have developed a new experimental model of an auxiliary liver with portal arterialization, avoiding the utilisation of the native hepatic hilium, necessary for the possible recovering of the proper liver in the case of a reversible fulminant hepatitis. PMID- 11891872 TI - Cortical vein end-to-end anastomosis after removal of a parasagittal meningioma. AB - A simple technique is described that facilitates a proper procedure of venous end to-end anastomosis during neurosurgical procedures. This technique, which consists of insertion of a Venflon tube in the vein during anastomosis, results in easier handling and proper apposition of the vein, resulting in an improved quality of the anastomosis. The technique was successfully applied in a patient after parasagittal meningioma resection, and the patency of the cortical vein was confirmed postoperatively on magnetic resonance venography. PMID- 11891873 TI - Restoration of fertility in oophorectomized rats after tubo-ovarian transplantation. AB - Improved microsurgical techniques for en bloc vascularized adnexal isograft in the rat is described. The graft of the right ovary together with its fallopian tube and upper third of uterus was transplanted orthotopically with end-to-side anastomoses between the donor aortic segment and recipient aorta and between the donor vena cava segment and recipient inferior vena cava, with end-to-end anastomosis of the donor and recipient uterus in a syngeneic, bilaterally oophorectomized rat. All transplantations were successful in terms of immediate vascular patency rate (10/10, 100%). Evidence of resumed ovarian function was obtained in 9 out of 10 rats (9/10, 90.0%) by histological demonstration of the vaginal smear, in which pregnancies were achieved in six rats (6/10, 60.0%) and six litters of healthy offspring were delivered 9 weeks later after transplantation. These results suggest that microsurgical ovarian transplantation provide a new and potential experimental model for the study of fertility restoration in humans. PMID- 11891874 TI - Microsurgical anterolateral thigh fasciocutaneous flap for facial contour correction in patients with hemifacial microsomia. AB - The correction of facial asymmetry in complex hemifacial microsomia presents a challenging problem for reconstructive surgeons. Numerous microsurgical flaps have been introduced for reconstruction of facial asymmetry. This article reports our experience in facial soft tissue reconstruction with microsurgical anterolateral thigh fasciocutaneous flap transfer in six patients with hemifacial microsomia. This flap, which has a reliable vascular pedicle and relatively thin pliable soft tissue, can provide an ideal treatment for facial asymmetry in hemifacial microsomia. PMID- 11891875 TI - Retrospective of the replantation of severed limbs in the People's Republic of China: current status and prospects. AB - Thirty-eight years have past since Chinese surgeons first reported the successful replantation of a severed limb. The following article reports the current status of replantation surgery in the People's Republic of China, addresses issues of complex replantation and reconstruction of severed limbs, and discusses future prospects and implications for replantation microsurgery. PMID- 11891877 TI - Systemic administration of 1R,4S-4-amino-cyclopent-2-ene-carboxylic acid, a reversible inhibitor of GABA transaminase, blocks expression of conditioned place preference to cocaine and nicotine in rats. AB - We examined the effect of 1R,4S-4-amino-cyclopent-2-ene-carboxylic acid (ACC), a reversible inhibitor of GABA transaminase, on the expression of conditioned place preference response to cocaine and nicotine in rats. Cocaine (20 mg/kg i.p.) and nicotine (0.4 mg/kg s.c.), but not vehicle or 300 mg/kg i.p. of ACC, produced a significant conditioned place preference response. Pretreatment of animals with 300 and 75 mg/kg i.p. of ACC significantly attenuated the expression of the cocaine- and nicotine-induced conditioned place preference responses, respectively. These results are the first to suggest that reversible inhibition of GABA transaminase may be useful in blocking cue-induced relapse to nicotine and cocaine. PMID- 11891878 TI - Differential distribution of 5HT2A and NMDA receptors in single cells within the rat medial nucleus of the solitary tract. AB - Activation of serotonin (5-hydroxytryptamine; 5HT) receptors of the 2A subtype (5HT2A) in the intermediate portion of the medial nucleus tractus solitarius (mNTS) produces marked hypotension and bradycardia. This portion of the mNTS receives major input from glutamatergic baroreceptor afferents. Thus, the cardiorespiratory effects of 5HT2A agonists may be attributed, in part, to interactions involving the glutamatergic target neurons, some of which express N methyl-D-aspartate (NMDA) glutamate receptors. To determine the functional sites for activation of 5HT2A receptors and their relationship to NMDA receptors in this region, we used electron microscopic immunocytochemistry for the localization of antipeptide antisera selectively recognizing each receptor protein in the intermediate mNTS in rat brain. Of 1,052 5HT2A-labeled profiles, 38% were dendrites and dendritic spines, 27% were unmyelinated axons, 14% were axon terminals, and 11% were glial processes. These 5HT2A-labeled profiles frequently contained NR1 gold particles with dendrites comprising 68% of the total dual-labeled profiles. In dendrites, the 5HT2A immunoreactivity was localized to cytoplasmic organelles or discretely distributed on synaptic or extrasynaptic segments of the plasma membrane. In contrast, NR1 immunoreactivity was prominently localized to postsynaptic junctions and these were distinct from the 5HT2A receptor labeling when coexpressed in the same dendrites. Dendrites containing both receptors composed 56% (224/399) of the total 5HT2A-labeled dendrites and 34% (224/659) of the total NR1-labeled dendrites. Our results provide the first ultrastructural evidence that in the intermediate mNTS, 5HT2A receptor agonists may affect the postsynaptic excitability of many of the same neurons that show NMDA-evoked responses to glutamate. PMID- 11891879 TI - Differential NR2A and NR2B expression between trigeminal neurons during early postnatal development. AB - N-methyl-D-aspartate (NMDA) receptors play an important role in the production of rhythmical trigeminal motor activity resembling suckling and chewing. The developmental relationship between the expression of NMDA receptor subunits and the function of neurons comprising brainstem oral-motor circuitry is not clear. We conducted receptor immunohistochemistry studies to demonstrate the expression of NR2A and NR2B subunits in trigeminal motoneurons (Mo5) and mesencephalic trigeminal neurons (Me5) during the first 2 weeks of development. During this time period, rats begin the transition from suckling to chewing, two distinct motor behaviors. In Mo5, NR2A and NR2B immunoreactivity was observed throughout the time frame sampled. A significant increase in the NR2A:NR2B ratio occurred between P3-4 and P11 due to a reduction in the number of NR2B immunoreactive neurons. The temporal and spatial expression of NR2A and NR2B was differentially regulated between caudal and rostral regions of Me5. In contrast to Mo5, the NR2A:NR2B ratio decreased between P0-1 and P11 in caudal Me5 due to a concurrent increase in the number of NR2A and NR2B immunoreactive neurons. In rostral Me5, NR2A and NR2B immunoreactivity emerged at P3 and P11, respectively. Our data provides further insight into the molecular changes of trigeminal neurons during the transition from suckling to chewing behaviors. The differences in the NR2A:NR2B ratio between Mo5 and Me5 suggest functional differences in these neurons during NMDA-mediated neurotransmission. PMID- 11891880 TI - Impaired preprodynorphin, but not preproenkephalin, mRNA induction in the striatum of mGluR1 mutant mice in response to acute administration of the full dopamine D(1) agonist SKF-82958. AB - Metabotropic glutamate receptor 1 (mGluR1) is highly expressed in striatonigral projection neurons of rat striatum. To define the role of mGluR1 in the regulation of striatal gene expression, the responsiveness of the three neuropeptide gene expression to a single injection of the dopamine D(1) agonist SKF-82958 was compared between mGluR1 mutant and wild-type control mice. We found that acute injection of SKF-82958 increased preprodynorphin (PPD), substance P (SP), and preproenkephalin (PPE) mRNAs in the dorsal and ventral striatum of mutant and wild-type mice in a dose-dependent manner (0.125, 0.5, and 2 mg/kg, i.p.) as revealed by quantitative in situ hybridization. However, the induction of PPD mRNA in both the dorsal and ventral striatum of mGluR1 minus sign/minus sign mice was significantly less than that of wild-type +/+ mice in response to the two higher doses of SKF-82958. In contrast to PPD, SP and PPE in the dorsal and ventral striatum of mGluR1 mutant mice were elevated to a similar level as that of wild-type mice. There were no differences in basal levels and distribution patterns of all three mRNAs between the two genotypes of mice treated with saline. These results indicate that mGluR1 selectively participates in striatonigral PPD induction in response to D(1) receptor stimulation. PMID- 11891881 TI - Studies of the biogenic amine transporters. 10. Characterization of a novel cocaine binding site in brain membranes prepared from dopamine transporter knockout mice. AB - Previous work suggested that the cocaine analog [(125)I]RTI-55 labels a novel binding site in rat brain membranes, which is not associated with the dopamine (DA), serotonin (5-HT), or norepinephrine (NE) transporters [Rothman et al. 1995 J Pharmacol Exp Ther 274:385-395]. Here, we tested whether this site is a product of the DA transporter (DAT) gene. We used a T-antigen knock-in at the DAT gene that results in an effective DAT knock-out (KO) confirmed by Southern blot, DAT immunohistochemistry, and [(125)I]RTI-55 ligand binding. Brain membranes were prepared from frozen whole brain minus caudate of wild-type (WT) B6/Sv129, +/+ and minus sign/minus sign (KO) mice. KO mice were used at approximately 23 days of age. Binding surface analysis of [(125)I]RTI-55 binding to membranes prepared from the brains of WT mice, with 100 nM citalopram to block binding to the 5-HT transporter (SERT), revealed two binding sites: the DAT and a second site, replicating previous studies conducted with rat brains. In the absence of the DAT (minus sign/minus sign mice), binding surface analysis demonstrated that [(125)I]RTI-55 labeled two sites: the NET and a second site called site "X." Structure-activity studies of site "X" demonstrated that high-affinity ligands for the DAT, NET, and SERT have low or negligible affinity for site "X." The relatively high density of site "X" in brain membranes and the fact that the K(i) values of cocaine and cocaethylene for site "X" are in the range achieved in the brain following cocaine administration suggests that site "X" could contribute to the pharmacological or toxicological effects of cocaine. Further progress in delineating the function of site "X" will depend on developing potent and selective agents for this site. PMID- 11891882 TI - Orexin B immunoreactive fibers and terminals innervate the sensory and motor neurons of jaw-elevator muscles in the rat. AB - The relationship between orexinergic terminals and the sensory and motor neurons of jaw-elevator muscles was examined by means of anti-orexin B (OXB) immunohistochemistry combined with horseradish peroxidase (HRP) retrograde tracing in the rat. HRP was initially injected into the jaw-elevator muscles; 48 h later the animals were sacrificed and fixed for HRP reaction and anti-OXB immunohistochemistry. OXB-like terminals were observed to distribute in the trigeminal mesencephalic nucleus (Vme) and the trigeminal motor nucleus (Vmo) where they closely contact the Vme neuronal somata and the Vmo neuronal somata and dendrites retrogradely labeled with HRP. The results of this study provide anatomical evidence of a direct OXB orexinergic innervation of the sensory and motor neurons controlling jaw-elevator muscles involved in mastication. Its functional significance related to the feeding behavior and bruxism is discussed. PMID- 11891883 TI - In vivo PET studies of the dopamine D1 receptors in rhesus monkeys with long-term MPTP-induced Parkinsonism. PMID- 11891886 TI - Palladium fluoride complexes: one more step toward metal-mediated C-F bond formation. AB - The first molecular complexes of palladium containing a Pd-F bond, both fluorides and bifluorides, were synthesized and fully characterized in the solid state and in solution. Reactivity studies of the Pd fluoride complexes revealed their unexpected stability and unusual chemical properties, different from the hydroxo, chloro, bromo, and iodo analogues. A novel efficient method to generate "naked fluoride" was developed using [(Ph(3)P)(2)Pd(F)Ph]. The naked fluoride from the Pd source fluorinated dichloromethane, deprotonated chloroform, and catalyzed di- and trimerization of hexafluoropropene under uncommonly mild conditions. PMID- 11891887 TI - [60]Fullerene as a substituent. AB - The substituent effect of the dihydro[60]fullerenyl group and its hydrophobic parameters have been evaluated quantitatively. The substituent constant has been determined from the pK value of a fullerene-based, para-substituted benzoic acid 1 in 80% dioxane/water (v/v) by NMR spectroscopy. The resulting Hammett sigma value of 0.06, consistent with a small electron-withdrawing effect of C(60), is a consequence of the fact that only inductive effects can be transmitted through the two tetracoordinate carbon atoms between the fullerene pi system and the para position of the benzoic acid moiety in 1. The parameter pi, which describes the hydrophobic character of the substituent C(60), has been evaluated as the difference between that of 1 and model compound 2. The pi value, which is larger than 3, indicates that the fullerene cage imparts high hydrophobicity to the molecule to which it is attached. Finally, we have evaluated how the fullerene spheroid influences the acid-base properties and nucleophilicity of the pyrrolidine nitrogen in a suitably functionalized fulleropyrrolidine. The fulleropyrrolidine 4 (pK(BD)(+)=5.6) is six orders of magnitude less basic and 1000 times less reactive than its model 3 (pK(BD)(+)=11.6). This may be related to through-space interactions of the nitrogen lone pair and the fullerene pi system. PMID- 11891888 TI - Silylanions: inversion barriers and NMR chemical shifts. AB - Alpha-substituent effects on inversion barriers and NMR chemical shifts have been studied on a set of silyl anions, [X(3-n)Y(n)Si](-) (X, Y=H, CH(3), and SiH(3)). The MP2/6-31+G* optimized structures show a pattern of increasing inversion barriers with augmenting numbers of methyl substituents. The highest barrier of 48.5 kcalmol(-1) is obtained for the (CH(3))(3)Si(-) ion. The silyl group displays the opposite effect by decreasing the inversion barrier to a minimum of 16.3 kcalmol(-1) in (SiH(3))(3)Si(-). The influence of counterions on these barriers is probed by addition of a lithium or potassium cation. In most cases, a decrease of the energy barriers with respect to the bare anions is observed. The (29)Si NMR chemical shifts calculated at the IGLO-DFT and GIAO-MP2 level of theory are also analyzed in view of the substituents and counterions. PMID- 11891889 TI - Molecular dynamics simulation of [Gd(egta)(H(2)O)](-) in aqueous solution: internal motions of the poly(amino carboxylate) and water ligands, and rotational correlation times. AB - Molecular dynamics simulations of [Gd(egta)(H(2)O)](-) (egta(4-)=3,12 bis(carboxymethyl)-6,9-dioxa-3,12-diazatetradecanedioate(4-)) have been performed without any artificial constraint on the first coordination sphere, such as covalent bonds between the Gd(3+) and the coordination sites. Two new crystallographic structures were determined for this gadolinium chelate and used to start two molecular dynamics simulations. [Gd(egta)(H(2)O)](-) and [Gd(egta)]( ) were both observed during the simulations, with a mean volume for the reaction of dissociation [Gd(egta)(H(2)O)](-)-->[Gd(egta)](-)+H(2)O of +7.2 cm(3)mol(-1), which corroborates the previously published experimental value of +10.5 cm(3)mol( 1). Changes in the conformation of the complex with the inversion of several dihedral angles are observed in the simulations independently from the water dissociation. Very fast changes of the third-order rotation axis direction of the Gd(3+) coordination polyhedron (of symmetry D(3h)) are observed during the simulations and are related to the mechanism of electronic relaxation of the complex. Different rotational correlation times (tau(R)) were calculated from the simulations on various observables of the complex. Protons of the inner sphere have different tau(R). The mean tau(R) of the two Gd-HW(HW=hydrogen of water molecule) vectors is 72% lower than tau(R) of the complex, and 75% lower than tau(R) of the vector Gd-OW (OW=oxygen of water molecule). This discrimination of the tumbling rates should be taken into account in future global (17)O NMR, EPR and NMRD (nuclear magnetic relaxation dispersion) data analysis. PMID- 11891890 TI - The impact of rigidity and water exchange on the relaxivity of a dendritic MRI contrast agent. AB - Variable-temperature, multiple magnetic field (17)O NMR, EPR and variable temperature (1)H nuclear magnetic relaxation dispersion (NMRD) measurement techniques have been applied to Gadomer 17, a new dendritic contrast agent for magnetic resonance imaging. The macromolecule bears 24 Gd(dota)-monoamide chelates (dota=N,N',N",N"'-tetracarboxymethyl-1,4,7,10-tetraazacyclododecane) attached to a lysine-based dendrimer. (17)O NMR and (1)H NMRD data were analysed simultaneously by incorporating the Lipari-Szabo approach for the description of rotational dynamics. The water exchange rate k(298)(ex)was found to be (1.0 +/- 0.1) x 10(6) s(-1), a value similar to those measured for other Gd(dota) monoamide complexes, and the activation parameters DeltaH++ =24.7 +/- 1.3 kJ mol( 1) and DeltaS++ = -47.4 +/- 0.2 JK(-1) mol(-1). The internal flexibility of the macromolecule is characterised by the Lipari-Szabo order parameter S(2)=0.5 and a local rotational correlation time tau(298)(l)= 760 ps, whereas the global rotational correlation time of the dendrimer is much longer, tau(298)(g)=3050 ps. The analysis of proton relaxivities reveals that, beside slow water exchange, internal flexibility is an important limiting factor for imaging magnetic fields. Electronic relaxation, though faster than in similar, but monomeric, Gd(III) chelates, does not limit proton relaxivity of this contrast agent (r(1)=16.5mM( 1)s(-1) at 298 K and 20 MHz). This analysis provides direct clues for the design of high-efficiency contrast agents. PMID- 11891891 TI - Zirconium-catalyzed amine oxidation: a mechanistic study. AB - The zirconium-catalyzed oxidation of amines in the presence of hydroperoxides gives the corresponding nitro compounds in high yields. In the present paper, we describe mechanistic details of this three-step oxidation, which was investigated by means of DFT calculations. It is shown that N-oxides, hydroxylamines, and nitroso derivatives are formed as intermediates. These compounds had already been postulated on the basis of synthetic experiments. During the oxidation process, the nitrogen atom changes its electronic character from a strong nucleophilic center to a moderate electrophilic center; this is reflected by the geometry of the transition states of the oxygen-transfer process. PMID- 11891892 TI - Electron counts for face-bridged octahedral transition metal clusters. AB - Kohn-Sham orbital energy patterns were used to rationalize valence electron counts for stable face-capped octahedral clusters [M(6)E(8)L(6)] (E=S, Se, Te, Cl; L=CO, PMe(3), Cl(-)). When L is a pi acceptor such as CO or PMe(3), stable closed-shell clusters are found for 80, 84, and 98 electrons. For L=Cl(-) (i.e. a pi-electron donor), only a count of 84 electrons appears favorable, as is found in [Mo(6)Cl(14)](2-). These counting rules apply to fivefold coordination of M, which becomes unstable if the electron count exceeds 98, for example, for M=Ni. In this case structures with tetrahedrally coordinated M are energetically favored, and this leads to different cluster structures. PMID- 11891893 TI - Ring currents in a proposed system containing planar hexacoordinate carbon, CB(2 )(6). AB - Current-density maps at the coupled Hartree-Fock level calculated in the CTOCD (continuous transformation of origin of current density) approach demonstrate the magnetic response of the hypothetical planar hexacoordinate carbon species, CB(2 )(6). In contrast with the empty CB(2-)(6) framework, which supports paramagnetic currents, the carbon-containing species has a typical diamagnetic pi-ring current that circulates undisturbed by the central atom. In spite of the unconventional nature of the species, the properties of 6pi CB(2-)(6) and 4pi CB(2-)(6) can be rationalised with the same orbital model that accounts for the diamagnetic pi current of benzene and the paramagnetic pi current of planar cyclooctatetraene. PMID- 11891894 TI - Ionized bicyclo[2.2.2]oct-2-ene: a twisted olefinic radical cation showing vibronic coupling. AB - The bicyclo[2.2.2]oct-2-ene radical cation (1(.+)) exhibits matrix ESR spectra that have two very different types of gamma-exo hydrogens (those hydrogens formally in a W-plan with the alkene pi bond), a(2H) about 16.9 G and a(2H) about 1.9 G, instead of the four equivalent hydrogens as would be the case in an untwisted C(2v) structure. Moreover, deuterium substitution showed that the vinyl ESR splitting is not resolved (and under about 3.5 G); this is also a result of the twist. Enantiomerization of the C(2) structures is rapid on the ESR timescale above 110 K (barrier estimated at 2.0 kcalmol(-1)). Density functional theory calculations estimate the twist angle at the double bond to be 11-12 degrees and the barrier as 1.2-2.0 kcalmol(-1). Single-configuration restricted Hartree-Fock (RHF) calculations at all levels that were tried give untwisted C(2v) structures for 1(.+), while RHF calculations that include configuration interactions (CI) demonstrate that this system undergoes twisting because of a pseudo Jahn-Teller effect (PJTE). Significantly, twisting does not occur until the sigma-orbital of the predicted symmetry is included in the CI active space. UHF calculations at all levels that include electron correlation (even semiempirical) predict twisting at the alkene pi bond because they allow the filled alpha and the beta hole of the SOMO to have different geometries. The 2,3-dimethylbicyclo[2.2.2]oct 2-ene radical cation (2(.+)) is twisted significantly less than 1(.+), but has a similar temperature for maximum line broadening. Neither the 2,3 dioxabicyclo[2.2.2]octane radical cation (3(.+)) nor its 2,3-dimethyl-2,3-diaza analogue (5(.+)) shows any evidence of twisting. Calculations show that the orbital energy gap between the SOMO and PJTE-active orbitals for 3(.+) is too large for significant PJTE stabilization to occur. PMID- 11891895 TI - Spectroscopy, electrochemistry, and molecular orbital calculations of metal-free tetraazaporphyrin, -chlorin, -bacteriochlorin, and -isobacteriochlorin. AB - Metal-free tetraazachlorin (TAC), -bacteriochlorin (TAB), and -isobacteriochlorin (TAiB) were characterized by electronic absorption, magnetic circular dichroism (MCD), fluorescence, and time-resolved ESR (TR-ESR) spectroscopy, and by cyclic voltammetry. The results are compared with those of metal-free tetraazaporphyrin (TAP). The potential difference DeltaE between the first oxidation and reduction couples decreases in the order TAP>TAiB>TAC>TAB. The splitting of both the Q and Soret bands decreases in the order TAB>TAC>TAP>TAiB. Corresponding to the split absorption bands, MCD spectra show a minus-to-plus pattern with increasing energy in both the Q and Soret regions, which suggests that the energy difference between the HOMO and second HOMO is larger than that between the LUMO and second LUMO. These spectroscopic properties and redox potentials were reproduced by molecular orbital calculations using the ZINDO/S Hamiltonian. The fluorescence quantum yields of the reduced species are much smaller than that of TAP. The zero field splitting (ZFS) parameters D and E of the excited triplet states (T1) of these species decrease and increase, respectively, on going from TAP to TAC and further to TAB. The D and E values of TAiB are larger than those of the other species. The results are supported by the absence of interaction between the spin over reduced pyrrole moieties of the HOMO and over the LUMO, and by calculations of ZFS under a half-point-charge approximation. PMID- 11891896 TI - Hydrophilic and lipophilic iron chelators with the same complexing abilities. AB - A new series of iron chelators with the same coordination sphere as the water soluble ligand O-trensox, but featuring a variable hydrophilic-lipophilic balance, have been obtained by grafting oxyethylene chains of variable length on a C-pivot tripodal scaffold. The X-ray structure of a ferric complex exhibiting tris(8-hydroxyquinolinate) coordination and solution thermodynamic properties (pK(a) of the ligands, stability constants of the ferric complexes) have been determined. The complexing ability (pFe(III) values) of the ligands are similar to that of O-trensox. Partition coefficients between water and octanol or chloroform have been measured and transport across a membrane has been mimicked ("shuttle process"). The results of biological assays (iron chelation with free ligands or iron nutrition with ferric complexes) could not be correlated with the partition coefficients. These results call into question the role of distribution coefficients (of the ligands and/or complexes) in the biological activities of iron chelators. PMID- 11891897 TI - Mixed copper-lanthanide metallomesogens. AB - This paper describes the first examples of heteropolynuclear metallomesogens that contain both a transition metal ion and a trivalent lanthanide ion. Adducts were formed between a mesomorphic [Cu(salen)] complex (salen=2,2'-N,N' bis(salicylidene)ethylenediamine) with six terminal tetradecyloxy chains and a lanthanide nitrate (Ln=La, Gd). Different stoichiometries were found, depending on the lanthanide ion: a trinuclear copper-lanthanum-copper complex [La(NO(3))(3)(Cu(salen))(2)] and a binuclear copper-gadolinium complex [Gd(NO(3))(3)Cu(salen)]. The compounds exhibit a hexagonal columnar mesophase (Col(H)) over a wide temperature-range with rather low melting temperatures. Although the clearing point could be observed for the parent [Cu(salen)] complex, the mixed f-d complexes decomposed in the high-temperature part of the mesomorphic domain before clearing. On the basis of X-ray diffraction measurements and molecular modelling, a structural model for the mesophase of the metal complexes is proposed. PMID- 11891898 TI - Exploring and expanding the three-dimensional structural diversity of supramolecular dendrimers with the aid of libraries of alkali metals of their AB(3) minidendritic carboxylates. AB - The synthesis of the alkali metal salts of 3,4,5-tris(n-alkan-1-yloxy)benzoic acid [(3,4,5)nG1-CO(2)M, where n is the number of methylenic units in the alkane group for n=12, 14, 16, 18 and M=Li, Na, K, Rb, Cs] is described. The structural analysis of these AB(3) molecules by a combination of methods which includes X ray diffraction experiments was performed. These experiments have demonstrated that (3,4,5)nG1-CO(2)M self-assemble at low temperatures into supramolecular cylinders and at high temperatures into spheres which subsequently self-organize into two-dimensional c2mm rectangular columnar, p6mm hexagonal columnar, three dimensional Pm(-)3n and Im(-)3m cubic lattices. In addition a novel unidentified liquid crystalline lattice was also discovered. The dependence between the symmetry of the lattice and the molecular structure of (3,4,5)nG1-CO(2)M was established. (3,4,5)nG1-CO(2)M represents one of the AB(3) minidendrons (i.e., first-generation monodendron attached to the periphery of larger generation dendrons) that is responsible for the control of the three-dimensional structures created from libraries of larger generations of dendrimers. Therefore, the molecular structure-lattice dependence elaborated here will aid the rational design of the three-dimensional shapes from larger generations of supramolecular dendrimers and of their lattices. In addition, the temperature responsive shape change of these supramolecular objects may generate new supramolecular concepts and technological applications. PMID- 11891899 TI - Preparation of a new chiral non-racemic sulfur-containing diselenide and applications in asymmetric synthesis. AB - The synthesis of the new chiral non-racemic sulfur-containing diselenide, di-2 methoxy-6-[(1S)-1-(methylthio)ethyl]phenyl diselenide, is described. When treated with ammonium persulfate this diselenide is transformed into the corresponding selenenyl sulfate, which acts as a strong electrophilic reagent and adds to alkenes, in the presence of methanol or water, to afford the products of selenomethoxylation or selenohydroxylation, respectively, with excellent diastereoselectivities. Starting from alkenes containing internal nucleophiles, asymmetric cyclofunctionalization reactions also resulted in good chemical yields, complete regioselectivities, and high diastereoselectivities. This sulfur containing diselenide can also be used in catalytic amounts to promote one-pot selenenylation-deselenenylation processes, from which several types of products can be obtained in high yield and with good enantiomeric excess. PMID- 11891900 TI - New and efficient chiral selenium electrophiles. AB - New chiral diselenides were prepared in a few steps from readily available starting materials. The selenium electrophiles generated from these diselenides were used for the efficient stereoselective inter- and intramolecular functionalization of alkenes. The substitution pattern influences the stereoselectivities and protection of the hydroxy moiety in the chiral side chain led to increased selectivities and yields in the selenenylation reactions. An additional substituent in the second ortho position was advantageous as well. Addition products with up to 96% de were obtained. The influence of the nucleophile on the outcome of selenenylations of alkenes was studied to some extent as well. PMID- 11891901 TI - Missing-link macrocycles: hybrid heterocalixarene analogues formed from several different building blocks. AB - The synthesis and structural characterization of hybrid heterocalix[4]arene analogues containing pyrrole, benzene, methoxy-substituted benzene, and pyridine subunits is described. Macrocycles 1 and 2, examples of calix[2]benzene[2]pyrrole and calix[1]benzene[3]pyrrole systems, respectively, are synthesized by the condensation of pyrrole and an appropriate phenylbis(carbinol). Macrocycles 3 and 7, examples of calix[2]benzene[1]pyridine[1]pyrrole and calix[1]pyridine[3]pyrrole, respectively, are synthesized by the use of a carbene based pyrrole-to-pyridine ring-expansion procedure. Single-crystal X-ray analysis reveals that compounds 1a, 1b, and 2b adopt 1,3-alternate conformations in the solid state, whereas compounds 3 and 7 display structures that are best described as "flattened partial cones" in terms of their conformation. (Series a refers to pure benzene-derived systems, whereas series b indicates macrocycles containing 5 methoxyphenyl subunits). In the solid state, the methoxy-functionalized macrocycles 1b and 2b, and the chloropyridine-containing macrocycle 7 exist as dimers. In the case of 1a and 7, these compounds interact with neutral solvent in the solid state. The conformations of the macrocycles in solution were explored by temperature-dependent proton NMR and NOESY spectral analysis. At 188 K, macrocycles 1a and 2a adopt flattened 1,3-alternate conformations, whereas macrocycles 3 and 7 exist in the form of flattened partial-cone conformations. Standard proton NMR titration analyses were carried out in the case of macrocycles 1a and 2a, and reveal that at least the second of these systems is capable of binding fluoride and chloride anions in CD(2)Cl(2) solution at room temperature (K(a)=571 and 17M(-1) in the case of 2a and F(-) and Cl(-), respectively). PMID- 11891902 TI - Fast interstrand cross-linking of cisplatin-DNA monoadducts compared with intrastrand chelation: a kinetic study using hairpin-stabilized duplex oligonucleotides. AB - The antitumor drug cisplatin forms two kinds of guanine-guanine cross-links with DNA: intrastrand, occurring mainly at GG sites, and interstrand, formed at GC sites. The former are generally more abundant than the latter, at least in experiments with linear duplex DNA. The formation of interstrand cross-links requires partial disruption of the Watson-Crick base pairing, and one could therefore expect the cross-linking reaction to be rather slow. In contrast with this expectation, kinetic measurements reported here indicate that interstrand cross-linking is as fast as intrastrand, or even faster. We have investigated the reactions between two hairpin-stabilized DNA duplexes, containing either a d(TGCA)(2) sequence (duplex TGCA) or a d(G(1)G(2)CA)-d(TG(3)CC) sequence (duplex GGCA), and the diaqua form of cisplatin, cis-[Pt(NH(3))(2)(H(2)O)(2)](2+), in an unbuffered solution kept at pH 4.5 +/- 0.1 and 20 degrees C. Using HPLC as the analytical method, we have determined the platination (first step) and chelation (second step) rate constants for these reaction systems. Duplex TGCA, in which the two guanines are quasi-equivalent, is found to be platinated very slowly (k=0.5 +/- 0.1M(-1)s(-1)) and to form the final interstrand cross-link very rapidly (k=13 +/- 3 x 10(-3) s(-11)). For GGCA, we find that G(1) is platinated rapidly (k=32 +/- 5M(-1)s(-1)) to form a long-lived monoadduct, which is only slowly chelated (k=0.039 +/- 0.001 x 10(-3) s(-1)) by G(2) (intrastrand), while G(2) is platinated one order of magnitude more slowly than G(1) (k=2.0 +/- 0.5M( 1)s(-1)) and chelated fairly rapidly both by G(1) (intrastrand: k=0.4 +/-0.1 x 10(-3) s(-1)) and G(3) (interstrand: k=0.2 +/- 0.1 x 10(-3) s(-1)); finally, G(3) is platinated at about the same rate as G(2) (k=2.4 +/- 0.5M(-1)s(-1)) and chelated very rapidly by G(2) (interstrand: k=10 +/- 4 x 10(-3) s(-1)). These results suggest that the low occurrence of interstrand cross-links in cisplatinated DNA is due to an extremely slow initial platination of guanines involved in d(GC)(2) sequences, rather than to a slow cross-linking reaction. PMID- 11891903 TI - Surface oxidation of carbon nanofibres. AB - Carbon nanofibres of the fishbone and parallel types were surface-oxidised by several methods. The untreated and oxidised fibres were studied with infrared spectroscopy, thermogravimetric analysis and X-ray photoelectron spectroscopy (XPS). Oxidation in a mixture of concentrated nitric and sulfuric acids proved to be the most effective method for creating oxygen-containing surface groups. This treatment results not only in the formation of carboxy and carboxyic anhydride groups, but also in the generation of ether-type oxygen groups between graphitic layers that are puckered at their edges. The IR spectroscopic data clearly show that the formation of oxygen-containing surface groups occurs at defect sites on the carbon nanofibres and that oxidation proceeds via carbonyl groups and other oxides to carboxy and carboxyic anhydride groups. Owing to the presence of defects, the two types of fibre have similar surface reactivities. With parallel nanofibres, in contrast to fishbone fibres, the macroscopic structure was severely affected by treatment with HNO(3)/H(2)SO(4). The HNO(3)/H(2)SO(4) treated fibres are highly wettable by water. PMID- 11891904 TI - Persistent bissilylated arenium ions. AB - A series of bissilylated arenium ions 1 with different substitution patterns on the aryl ring have been synthesized by hydride abstraction from 2-aryl substituted 2,6-dimethyl-2,6-disilaheptanes (2) via transient silylium ions. The arenium ions have been identified by their characteristic NMR chemical shifts, (delta(29)Si=19.1-25.6, delta(13)C(ipso) =89.0-102.4, delta(13)C(ortho)=160.9 182.0, delta(13)C(meta)=132.5-146.9, delta(13)C(para)=150.2-169.9) supported by quantum mechanical calculations of structures, energies, and magnetic properties at the B3LYP/6-311G(d,p)//B3LYP/6-31G(d) + DeltaZPVE level of theory. The calculations clearly reveal the charge dispersing and stabilizing effect of the silyl substituents in arenium ions 1. The bissilylated benzenium ion 1a is more stable than the parent benzenium ion (C(6)H(7)(+)) by 37.6 kcalmol(-1). The synthesized arenium ions 1 are stable in solution at room temperature for periods ranging from a few hours to days. This unusual stability is attributed to: 1) the thermodynamic stabilization of the arenium ion by two beta-silyl substituents and 2) the essentially non-nucleophilic reaction conditions (the use of the weakly coordinating [B(C(6)F(5))(4)](-) anion and aromatic hydrocarbons as solvents). Addition of stronger nucleophiles than aromatic hydrocarbons (for example, acetonitrile) results in desilylation of the arenium ion 1 and recovery of the 2 aryl-2,6-disilaheptane moiety. PMID- 11891905 TI - O-H...O interactions involving doubly charged anions: charge compression in carbonate-bicarbonate crystals. AB - The O-H...O interaction formed by the anions HCO(3)(-) and CO(3)(2-) has been investigated on the basis of data retrieved from the Inorganic Crystal Structure Database (ICSD) and by means of ab initio computations. It has been shown that the O-H...O separations associated with HCO(3)(-)...(3)(2-) interactions are shorter than those found in crystals containing hydrogen carbonate monoanions such as HCO(3)(-)...HCO(3)(-). Ab initio MP2/6-311G++(2d,2p) computations on the crystal Na(3)(HCO(3))(CO(3)).2H(2)O have shown that the interaction between the monoanion donor and the dianion acceptor, for example HCO(3)(-)...CO(3)(2-), is more repulsive than that between singly charged ions, for example HCO(3)( )...HCO(3)(-), but is largely overcompensated for by anion-cation electrostatic attractions. The shortening of the (-)O-H...O(2-) interaction relative to the ( )O-H...O(-) interaction has been explained as a consequence of the increased charge compression, that is of the stronger cation-anion interactions established by the CO(3)(2-) dianions with respect to those established by monoanions, and does not reflect an increase in the strength of the (-)O-H ...O(-) interaction. To expand the structural sample in the crystal packing analysis, the structure of the novel mixed salt K(2)Na(HCO(3))(CO(3)).2H(2)O has been determined by single crystal X-ray diffraction and compared with the structure of the salt Na(3)(HCO(3))(CO(3)).2H(2)O used in the computations. PMID- 11891906 TI - Porphyrin-based peptide receptors: syntheses and NMR analysis. AB - The synthesis and purification of a water-soluble host compound that contains three pyridinium units and one spacer-connected benzocrown ether unit in the meso positions of porphyrin and of its Zn(II) or Cu(II) complexes is described. Metalation leads to small (compared to the apo-derivative) changes of selectivities with different peptides, with complexation constants in water of above 10(5)M(-1). One complex containing the tripeptide Gly-Gly-Phe is analyzed in detail by COSY, HSQC, HMBC, and NOESY NMR experiments. Temperature-dependent spectra show activation energies for a intramolecular hydrogen exchange of amide protons with valence isomerization of the porphyrin ring, in accordance with the literature. Sharp signals for the spin system are only found at elevated temperature. Vicinal coupling constants within the crown ether moiety indicate stronger puckering than that reported for benzocrowns. All NMR signals of the complexed peptide are shielded, in particular those of the terminal phenylalanine unit, in line with its stacking on the porphyrin surface. A corresponding structural model, obtained by CHARMm simulation, is also in line with the observed intermolecular NOE cross peaks. PMID- 11891907 TI - The trifluoromethoxycarbonyl radical CF(3)OCO. AB - The trifluoromethoxycarbonyl radical CF(3)OCO is formed by low-pressure flash pyrolysis of CF(3)OC(O)OOC(O)OCF(3) or CF(3)OC(O)OOCF(3) in the presence of a high excess of CO and subsequent quenching of the reaction mixture as a CO matrix. The IR and UV spectra are recorded, and a DFT study of CF(3)OCO is presented. According to the quantum chemical calculations, two rotamers should exist with an energy difference between the isomers equal or larger than 12 kJmol(-1). By comparing calculated and observed IR spectra, the presence of the trans form of the CF(3)OCO radical is identified in the matrix. The reaction of CF(3)O radicals with CO leading to CF(3)OCO is calculated to be exothermic by 33.6 kJmol(-1). CF(3)OCO dissociates when irradiated by UV light with lambda<370 nm into CF(3) radicals and CO(2). Experiments show that CF(3) radicals do not react with solid CO to give CF(3)CO. PMID- 11891909 TI - Organometallic chemistry of fluorinated allenes. AB - Reaction of 1,1-difluoroallene and tetrafluoroallene with a series of transition metal complex fragments yields the mononuclear allene complexes [CpMn(CO)(2)(allene)] (1), [(CO)(4)Fe(allene)] (2), [(Ph(3)P)(2)Pt(C(3)H(2)F(2))] (4), [Ir(PPh(3))(2)(C(3)H(2)F(2))(2)Cl] (5), and the dinuclear complexes [mu eta(1)-eta(3)-C(3)H(2)F(2))Fe(2)(CO)(7)] (3), [Ir(PPh(3))(C(3)H(2)F(2))(2)Cl](2) (6), and [mu-eta(2)-eta(2)-C(3)H(2)F(2))(CpMo(CO)(2))(2)] (9), respectively. In attempts to synthesize cationic complexes of fluorinated allenes [CpFe(CO)(2)(C(CF(3))=CH(2))] (7a), [CpFe(CO)(2)(C(CF(3))=CF(2))] (7b) and [mu-I (CpFe(CO)(2))(2)][B(C(6)H(3)-3,5-(CF(3))(2))(4)] were isolated. The spectroscopic and structural data of these complexes revealed that the 1,1-difluoroallene ligand is coordinated exclusively with the double bond containing the hydrogen substituted carbon atom. 1,1-Difluoroallene and tetrafluoroallene proved to be powerful pi acceptor ligands. PMID- 11891910 TI - Stereoelectronic substituent effects in polyhydroxylated piperidines and hexahydropyridazines. AB - From the pK(a) values of the conjugate acids of a large series of hydroxylated piperidines and hexahydropyridazines, a consistent difference in basicity was found between stereoisomers having an axial or equatorial hydroxyl (OH) group either beta or gamma to the amine. Compounds with an equatorial OH group in the 3 position were 0.8 pH units more acidic than otherwise identical compounds with an axial OH group, whilst compounds with an equatorial OH group in the 4-position relative to the amine were 0.4 pH units more acidic than the corresponding compound with an axial OH. A similar effect was observed for the COOMe substituent. The difference in electron-withdrawing power of axial and equatorial substituents was explained by a difference in charge-dipole interactions in the two systems. Since this stereoelectronic substituent effect causes differences in basicity in different conformers, certain piperidines and hexahydropyridazines were found to change conformation upon protonation. A method for predicting the pK(a) of piperidines which takes stereochemistry into account is described. PMID- 11891908 TI - Sulfide oxidation by hydrogen peroxide catalyzed by iron complexes: two metal centers are better than one. AB - Peroxoiron species have been proposed to be involved in catalytic cycles of iron dependent oxygenases and in some cases as the active intermediates during oxygen transfer reactions. The catalytic properties of a mononuclear iron complex, [Fe(II)(pb)(2)(CH(3)CN)(2)] (pb=(-)4,5-pinene-2,2'-bipyridine), have been compared to those of its related dinuclear analogue. Each system generates specific peroxo adducts, which are responsible for the oxidation of sulfides to sulfoxides. The dinuclear catalyst was found to be more reactive and (enantio)selective than its mononuclear counterpart, suggesting that a second metal site affords specific advantages for stereoselective catalysis. These results might help for the design of future enantioselective iron catalysts. PMID- 11891911 TI - Supramolecular polymers generated from heterocomplementary monomers linked through multiple hydrogen-bonding arrays--formation, characterization, and properties. AB - Supramolecular polymers are described that are derived from the association of two homoditopic heterocomplementary monomers through sextuple hydrogen-bonding arrays. They form fibers and a variety of different materials depending on the conditions. The strong affinity of the DAD-DAD (D=donor, A=acceptor) hydrogen bonding sites for double-faced cyanuric acid type wedges drives the supramolecular polymeric assembly in apolar and chlorinated organic solvents. The marked influence of stoichiometry, as well as end-capping and cross-linking agents upon fiber formation is revealed in solution and by electron microscopy (EM). The results further contribute to the development of a supramolecular polymer chemistry that comprises reversible polymers formed through recognition controlled noncovalent connections between the molecular components. Such materials are, by nature, dynamic and present adaptive character in view of their ability to respond to external stimuli. PMID- 11891913 TI - Developmentally-regulated changes of Xist RNA levels in single preimplantation mouse embryos, as revealed by quantitative real-time PCR. AB - Xist RNA localizes to the inactive X chromosome in cells of late cleavage stage female mouse embryos (Sheardown et al., 1997: Cell 91:99-107). Fluorescence in situ hybridization (FISH), however, does not quantify the number of Xist transcripts per nucleus. We have used real-time reverse transcription-polymerase chain reaction (RT-PCR) to measure Xist RNA levels in single preimplantation embryos and to establish developmental profiles in both female and male samples. The gender of each embryo was readily established based on Xist RNA levels, by counting Xist gene copies per cell, and by independent detection of the presence/absence of Sry, a Y chromosome-specific gene. Xist expression in males was found to be very low at all stages, as suggested by FISH. In contrast, female embryos contained measurable levels of Xist mRNA starting at the late 2-cell stage and rapidly accumulated Xist transcripts until morula stage. Xist RNA accumulation per embryo then reached a plateau, while cell division continued. We propose that during early cleavage high enough levels of Xist mRNA are transcribed to generate a pool of unbound molecules. This pool would serve to temporarily maintain X chromosome inactivation without additional transcription while the trophectoderm and inner cell mass (ICM) differentiate. The ICM would then loose the paternally imprinted pattern of X inactivation originally present in all embryonic cells. PMID- 11891914 TI - On the regulation of Crisp-1 mRNA expression and protein secretion by luminal factors presented in vivo by microperfusion of the rat proximal caput epididymidis. AB - Synthesis and secretion of certain epididymal proteins are regulated by lumicrine factors from the testis or from upstream regions of the excurrent ducts. Cysteine rich secreted protein-1 (Crisp-1) is a major androgen regulated protein in the epididymal lumen fluid of the rat and other species. Previous research has demonstrated that disturbance of the luminal microenvironment through obstruction of the tract reduces Crisp-1 synthesis and secretion. The present study was undertaken to determine the influence of the luminal microenvironment on rat proximal caput epididymal Crisp-1 secretion into lumen fluid and on Crisp-1 gene expression in the same tubules. Western blot analysis demonstrated that Crisp-1 protein concentrations were reduced from control levels by perfusion with artificial caput fluid containing no testicular factors and were not increased by perfusion with fluids containing rete testis fluid proteins. Crisp-1gene expression was also reduced by perfusion with artificial caput fluid and not increased by perfusion with rete testis fluid proteins. Perfusion with artificial caput fluid containing 5alpha-dihydrotestosterone did increase one Crisp-1 transcript. This study demonstrates that intraluminal testicular proteins are not important co-regulators with androgens of Crisp-1gene expression or resulting Crisp-1 secretion into the rat proximal caput tubule lumen in vivo. PMID- 11891915 TI - Expression of p53 in luminal and glandular epithelium during the growth and regression of rat uterus during the estrous cycle. AB - It has been well recognized that epithelial cells of the rat endometrium cyclically proliferate and die during the estrous cycle. The aim of the present study was to determine p53 expression pattern and correlate it with the the apoptotic pattern of epithelial cells of the rat uterus during the estrous cycle. The p53 mRNA and protein expression pattern was assessed by in situ hybridization and immunohistochemistry. The apoptotic index was determined by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and electron microscopy. The highest p53 mRNA content, detected by in situ hybridization, was observed on the metestrus day both in the luminal and the glandular epithelia. During this period both epithelia presented high proliferation. The content of p53 mRNA markedly decreased in the following days, presenting its minimal values on the estrus day. The highest number of p53 immunopositive nuclei, in both the luminal and the glandular epithelia, was also detected on the metestrus day, while the lowest one was found on estrus day. On the proestrus day, p53 protein was predominantly detected in the glandular epithelium. However, on the estrus day, p53 protein was detected both in the nuclei and in the cytoplasm of luminal epithelial cells, predominantly in the cytoplasm. The highest apoptotic index in both the luminal and the glandular epithelia was observed on the estrus day whereas the lowest one was observed on the proestrus day. The apoptotic index values were higher in the luminal than in the glandular epithelia. The overall results indicate that p53 expression at both mRNA and protein levels is higher on the metestrus day when the apoptotic index is low. This suggests that p53 should play an important physiological role during proliferative phases of the estrous cycle in the rat uterus. PMID- 11891916 TI - Subtilisin proprotein convertase-6 expression in the mouse uterus during implantation and artificially induced decidualization. AB - During implantation, a balance of factors regulates the invasive properties of the embryo and the anti-invasive properties of uterine decidua. Although antiproteinases such as the metalloproteinase inhibitor TIMP-3 are thought to play critical roles in preventing the overaggressive invasion of trophoblasts, the mechanism of antiproteinase regulation is unknown. Recently, the prohormone convertase SPC-6 has been found to be co-expressed in embryo-proximal decidua in association with TIMP-3. As members of this serine proteinase family are known to activate latent TGFbeta family members which regulate decidual TIMP-3 levels, we sought to characterize the expression of SPC-6 during pregnancy and artificial decidualization. In this study, we demonstrate that the zone of SPC-6 gene expression exhibits a great degree of temporal and spatial overlap with TIMP-3 gene expression in uterine decidua from E5.5 through to E8.5. Like TIMP-3, we demonstrate that SPC-6 expression is induced during the decidual cell response using an in vivo model of artificial decidualization. Both the secreted and membrane bound forms of SPC-6 are expressed throughout the period of decidualization, suggesting that SPC-6 may play multiple roles during this developmental period. This is confirmed by our observation of the movement of SPC 6 expression to the presumptive placental region, as TIMP-3 expression regresses at the implantation site. PMID- 11891917 TI - In vitro formation of tetraploid rat blastocysts after fusion of two-cell embryos. AB - Gene targeting technology is not available in the rat which is an animal model of major importance, e.g., in cardiovascular research. This is due to the fact that the rat embryonic stem cell (ESC)-like cells established by several groups do not form germ-line chimeras when injected into blastocysts. In the mouse, the aggregation of ESC with tetraploid embryos has allowed the generation of animals completely derived from these cells. However, aggregation of rat ESC-like cells with tetraploid rat embryos has not yet been attempted to evaluate their developmental capacity. Therefore, we established a method to produce tetraploid rat embryos by fusion at the two-cell stage. Chemical fusion by polyethylene glycol (PEG) was shown to be less efficient (56.3% fused embryos) than electrofusion (96.1% fused embryos). The rate of development of fused embryos to blastocysts was independent of the fusion method and similar to the rate of control embryos. However, this rate was lower when the embryos had been cultured from the zygote state before fusion (14-20%) compared to freshly isolated two cell embryos (41-63%). Alike for the mouse, blastocysts derived from fused two cell rat embryos contained about half the number of cells as control blastocysts and were homogeneously tetraploid with no evidence of mosaicism. This method may be useful for the establishment of gene-targeting technology in the rat. PMID- 11891918 TI - Characteristics of the cell membrane fluidity, actin fibers, and mitochondrial dysfunctions of frozen-thawed two-cell mouse embryos. AB - Physical and chemical alterations caused by the freezing and thawing and their effects on survivals/developments in vitro were investigated. Of a total of 452 two-cell mouse embryos, the overall survival rate of the frozen-thawed embryos was 76.1% (344/452). The blastocyst formation of the frozen-thawed embryos was 32.6% (44/136) compared to 74.5% (117/157) in the fresh embryos (P<0.05). The total number of cells in a blastocyst also decreased from 96.0 +/- 19.0 (n=26) in the fresh embryos to 42.0 +/- 11 .34 (n=30) in the frozen-thawed embryos (P<0.05). Fluorescence recovery after photobleaching (FRAP) measurement revealed about 5-fold decrease in the cell membrane fluidity with a characteristic time constant (tau) of 1.46 +/- 0.13 sec (n=5) in the frozen-thawed embryos as opposed to 0.28 +/- 0.04 sec (n=5) in the fresh embryos (P<0.05). The relative amount of H(2)O(2) in an embryo as quantified by the fluorescence intensity of 2',7' dichlorofluorescein (DCF) showed 62.8 +/- 23.5 (n=24) and 34.2 +/- 14.5 (n=20) in the frozen-thawed embryos and in the fresh embryos, respectively (P<0.05). The distribution of actin filaments in the frozen-thawed embryos revealed an uneven distribution, particularly discontinuities at the "actin band," which contrasted to an even distribution shown in the fresh embryos. Mitochondrial staining by Rhodamine 123 showed that there was no significant difference between the two treatments in the number and in the distribution of viable mitochondria, but a marked aggregation was seen in the arrested embryos. No Annexin V binding was detected in two-cell or four-cell embryos while the binding was positive in the arrested embryos. The mitochondrial membrane potential measured by a membrane potential-sensitive fluorescent probe 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazol- carbocyanine iodide (JC-1) revealed a marked depolarization in the frozen-thawed embryos. Finally, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end-labeling (TUNEL) was employed to quantify the DNA fragmentation. In 75.0% cells of blastocysts (n=24) in the frozen-thawed embryos, the DNA fragmentation was detected as opposed to 37.0% in the fresh embryos (n=20) (P<0.05). Taken together, it is proposed that during the cryopreservation, two-cell mouse embryos are subjected to physical and chemical alterations, including destruction of the cell membrane integrity, redistribution of actin fibers, mitochondrial depolarizations, and increased reactive oxygen species (ROS) productions, which then may trigger the apoptotic cascade leading to a decrease in the survival rate and in the developmental rate of the embryos. PMID- 11891919 TI - Nucleolar protein allocation and ultrastructure in bovine embryos produced by nuclear transfer from granulosa cells. AB - In the present study immunofluorescence confocal laser scanning microscopy, autoradiography following (3)H-uridine incubation and transmission electron microscopy were used to evaluate the nucleolar protein localization, transcriptional activity, and nucleolar ultrastructure during genomic re programming in bovine embryos reconstructed by nuclear transfer from granulosa cells into non-activated cytoplasts followed by activation. During the 1st cell cycle (1-cell embryos), no autoradiographic labelling was detected. Ultrastructurally, nucleoli devoid of a granular component were observed. During the 2nd cell cycle (2-cell embryos) autoradiographic labelling was also lacking and the embryos displayed varying degrees of nucleolar inactivation. During both the 3rd (4-cell embryos) and 4th (tentative 8-cell embryos), cell cycles autoradiographic labelling was lacking in some embryos, while others displayed labelling and associated formation of fibrillo-granular nucleoli. During the 5th cell cycle (tentative 16-cell embryos), all embryos displayed autoradiographic labelling and fibrillo-granular nucleoli. In some blastomeres, however, deviant nucleolar ultrastructure was observed. During the first cell cycle labelling of RNA polymerase I, fibrillarin, upstream binding factor (UBF) and nucleolin (C23) was localized to nuclear entities. During the 2nd cell cycle, only labelling of RNA polymerase I and fibrillarin persisted. During the 3rd and 4th cell cycle labelling of fibrillarin persisted, labelling of nucleophosmin (B23) appeared and that of nucleolin re-appeared. During the 5th cell cycle almost all embryos showed complete labelling of all proteins except for UBF, which lacked in more than half of the embryos. In conclusion, bovine granulosa cell nuclear transfer embryos showed re-modelling of the nucleoli to an inactive form followed by re formation of fibrillo-granular nucleoli. The re-formation of fibrillo-granular nucleoli was initiated already during the 3rd cell cycle, which is one cell cycle earlier than in in vivo- and in vitro-derived bovine embryos. Moreover, in more than half of the embryos, UBF could not be immunocytochemically localized to the nucleolar compartment during the 5th cell cycle indicating lack of developmental potentials. PMID- 11891921 TI - Effect of extracellular matrix proteins on in vitro testosterone production by rat Leydig cells. AB - The aim of this study was to detect the effect of extracellular matrix (ECM) proteins on rat Leydig cell shape, adhesion, expression of integrin subunits and testosterone production, in vitro. Leydig cells isolated from adult rats were cultured on plates uncoated or coated with different concentrations of laminin-1, fibronectin, or type IV collagen in the presence or absence of hCG for 3 or 24 hr. A significant increase of cell adhesion and of alpha3, alpha5, and beta1 integrin subunit expression was observed when cells were cultured on ECM proteins, compared to those grown on uncoated plates. Leydig cells cultured on glass coverslips coated with ECM proteins for 24 hr exhibited elongated shapes with long cell processes (spreading), while cells cultured on uncoated plates showed few cell processes. A significant decrease in testosterone production was observed when basal and hCG-stimulated Leydig cells were cultured for 3 or 24 hr on plates coated with type IV collagen (12 and 24 microg/cm(2)) compared to uncoated plates. A significant though a slighter decrease in testosterone production was also observed in cells cultured on plates coated with fibronectin (12 and 24 microg/cm(2)), compared to uncoated plates. Laminin-1 did not modify testosterone production under basal or hCG stimulated conditions. These results suggest that ECM proteins are able to modulate Leydig cell steroidogenesis, in vitro. PMID- 11891920 TI - Effect of elevated Ca(2+) concentration in fusion/activation medium on the fusion and development of porcine fetal fibroblast nuclear transfer embryos. AB - The present study examined the effect of elevated Ca(2+) concentration in fusion/activation medium on the fusion and development of fetal fibroblast nuclear transfer (NT) porcine embryos. Frozen-thawed and serum starved fetal fibroblasts were transferred into the perivitelline space of enucleated oocytes. Cell fusion and activation were induced simultaneously with electric pulses in 0.3 M mannitol-based medium containing 0.1 or 1.0 mM CaCl(2). Some fused embryos were further activated 1 hr after the fusion treatment by exposure to an electric pulse. The NT embryos were cultured in vitro for 6 days. Fusion and blastocyst formation rates were significantly (P<0.05) increased by increasing the Ca(2+) concentration from 0.1 mM (67.1 and 6.3%) to 1.0 mM (84.7 and 15.8%). However, no difference in the number of cells in blastocysts was observed between the two groups. A higher percentage of blastocyst was also observed when control oocytes were parthenogenetically activated in the presence of elevated Ca(2+) (19.3% vs. 32.4%, P<0.05). When the reconstituted oocytes were fused in the medium containing 1.0 mM CaCl(2), increasing the number of pulses from 2 to 3 or an additional activation treatment did not enhance the blastocyst formation rate or cell number in blastocysts. These results demonstrate that increasing the Ca(2+) concentration in the fusion/activation medium can enhance the fusion and blastocyst formation rates of fetal fibroblast NT porcine embryos without an additional activation treatment. PMID- 11891922 TI - Nitric oxide-mediated inhibition of follicular apoptosis is associated with HSP70 induction and Bax suppression. AB - Nitric oxide (NO) has recently emerged as a potential regulator of follicular development because of its involvement in the regulation of several physiological functions of the ovary. NO influences apoptotic cell death of follicular cells as a follicle survival factor. The present study was conducted (1) to investigate the mechanism involved in the protective effect of NO on spontaneously induced follicular apoptosis in serum-free condition and (2) to determine the role of NO on the expression of mRNAs and proteins for HSP70 and Bax. Preovulatory follicles obtained from PMSG-primed rats were cultured for 24 hr in serum-free medium with or without sodium nitroprusside (SNP), a NO generator. Granulosa cells within follicles incubated in medium alone for 24 hr exhibited extensive apoptosis. Treatment of SNP in the culture medium blocked this onset of apoptosis. Both mRNA and protein levels of HSP70 were highly increased with SNP than those of control group. On the contrary, those of Bax were suppressed with SNP treatment. Results of the present study suggest that NO prevents rat preovulatory follicular apoptosis in vitro by stimulating HSP70 and suppressing Bax expression. PMID- 11891924 TI - Protamine 1: protamine 2 stoichiometry in the sperm of eutherian mammals. AB - We have compared the relative proportion of protamine 1 (P1) and protamine 2 (P2) bound to DNA in the sperm of a variety of eutherian mammals to obtain insight into how these two proteins interact in sperm chromatin. Gel electrophoresis (combined with microdensitometry) and high performance liquid chromatography (HPLC) were used to determine the content of the two protamines, and the identity of each protein was confirmed by amino-terminal sequencing or amino acid analysis. The sperm of all species examined contained P1, but P2 was found to be present only in certain species. Unlike the fixed ratio of core histones that package DNA into nucleosomes in all somatic cells, the proportion of P2 present in mature sperm was found to be continuously variable from 0 to nearly 80%. These results show that P1 and P2 do not interact with each other or DNA to form a discrete complex or subunit structure that is dependent upon particular P1/P2 stoichiometries. Data obtained from a number of closely and distantly related species also indicate that while the P2 content of sperm chromatin is allowed to vary over a wide range during the course of evolution, the relative proportion of P1 and P2 are tightly regulated within a genus. PMID- 11891925 TI - Localization of plasminogen in the extracellular matrix of hamster eggs: exogenous activation by streptokinase. AB - The plasminogen activator (PA)/plasminogen/plasmin proteolytic system has begun to be taken into account in the fertilization process. In this study, we demonstrated the presence of plasminogen in the extracellular matrix (ECM) of hamster oocytes by indirect immunofluorescence and immunoperoxidase assays using human anti-plasminogen. Plasminogen appeared first on the zona pellucida (ZP) of ovarian oocytes and later on the plasma membrane (PM) of oviducal eggs. This would suggest that oviducal oocytes modulate the expression of plasminogen binding sites on the PM. Human plasminogen as well as that of other species, known to be activated by streptokinase (SK), is rapidly converted to a plasmin-SK complex. We demonstrated the rapid formation of a SK-plasminogen complex that yields plasmin in the blood plasma of hamsters. Both the in vivo and in vitro SK treatment of eggs from superovulated female hamsters caused a decreased in the ZP dissolution time (ZPdt), probably either due to the proteolytic effect of plasmin or due to the SK-Plasminogen. Extracellular proteolysis assays carried out on agar-casein plates confirmed the proteolytic activity of SK-incubated eggs; the controls, on the contrary, failed to display a halo. These studies show that (1) superovulated hamster eggs contain plasminogen in their ECM, (2) oviducal eggs exhibit plasminogen on their PMs, indicating the presence of their corresponding binding sites, (3) in hamsters, SK, a non-enzymatic exogenous protein would be capable of activating ECM plasminogen to plasmin, and (4) the complex SK plasminogen and/or the plasmin are capable of changing the ZPdt with alpha chymotrypsin. PMID- 11891923 TI - Inhibition of apoptosis in vitellogenic ovarian follicles of rainbow trout (Oncorhynchus mykiss) by salmon gonadotropin, epidermal growth factor, and 17beta estradiol. AB - We have examined the ability of selected hormones and growth factors to suppress the spontaneous onset on apoptotic DNA fragmentation in isolated vitellogenic rainbow trout ovarian follicles cultured in serum-free conditions. Primary culture of isolated follicles for 24 hr in serum-free conditions resulted in a 3 5-fold increase in the amount of fragmented DNA as compared to non-cultured controls, measured by radioactive 3'end-labeling. Culture in medium containing salmon gonadotropin (SG-G100; 1, 5 microg/ml) suppressed the spontaneous onset of DNA fragmentation in dose-dependent fashion. Culture with 1 ng/ml 17beta estradiol, or 100 ng/ml epidermal growth factor also suppressed the spontaneous onset of apoptosis, whereas culture with higher concentrations of 17beta estradiol (10 and 100 ng/ml), insulin-like growth factor I (IGF-I; 1, 10, and 100 ng/ml), or 8-bromo-cAMP (0.1, 1, and 5 mM) was ineffective in suppressing apoptosis. Apoptosis was confirmed as the mode of cell death through positive identification of nuclear morphological characteristics associated with apoptosis, and positive staining for fragmented DNA using in situ end-labeling (TUNEL); apoptotic cells identified in situ were almost exclusively localized to the thecal/epithelial region of the follicle. In summary, this study shows that vitellogenic ovarian follicles are susceptible to apoptosis and that both endocrine and locally-derived growth factors may play a role as cell survival factors by preventing apoptosis. The study also suggests that rainbow trout differ markedly from mammals both in terms of the cell types susceptible to apoptosis and the responsiveness to specific growth factors in terms of inhibiting apoptosis. PMID- 11891926 TI - Receptor mediated yolk protein uptake in the crab Scylla serrata: crustacean vitellogenin receptor recognizes related mammalian serum lipoproteins. AB - The receptor-mediated uptake of major yolk protein precursor, vitellogenin (Vg) is crucial for oocyte growth in egg laying animals. In the present study plasma membrane receptor for Vg was isolated from the oocyte of the red mud crab, Scylla serrata. Vitellogenin receptor (VgR) protein was visualized by ligand blotting using labeled crab Vg ((125)I-Vg) as well as labeled low density lipoprotein ((125)I -LDL) and very low density lipoprotein ((125)I-VLDL) isolated from rat. The endocytosis of Vg was visualized in the crab oocyte by ultrastructural immunolocalization of Vg. The Vg receptor was purified by gel filtration high performance liquid chromatography (HPLC) and its molecular weight was estimated to be 230 kDa. In direct binding studies, the receptor exhibited high affinity (dissociation constant K(d) 0.8x10(minus sign6) M) for crab Vg. Vitellogenin receptor was observed to have an increased affinity to crab Vg in the presence of Ca(2+) and the binding was inhibited by suramin, suggesting similarities between crab VgR and low density lipoprotein receptor (LDLR) superfamily of receptor protein. Furthermore, the crab VgR showed significant binding ability to mammalian atherogenic lipoproteins such as LDL and VLDL. This suggests that there is a tight conservation of receptor binding sites between invertebrate (crab) Vg and vertebrate (rat) LDL and VLDL. PMID- 11891928 TI - Evidence that multifunctional calcium/calmodulin-dependent protein kinase II (CaM KII) participates in the meiotic maturation of mouse oocytes. AB - Calcium-dependent signaling pathways are thought to be involved in the regulation of mammalian oocyte meiotic maturation. However, the molecular linkages between the calcium signal and the processes driving meiotic maturation are not clearly defined. The present study was conducted to test the hypothesis that the multi functional calcium/calmodulin-dependent protein kinase II (CaM KII) functions as one of these key linkers. Mouse oocytes were treated with a pharmacological CaM KII inhibitor, KN-93, or a peptide CaM KII inhibitor, myristoylated AIP, and assessed for the progression of meiosis. Two systems for in vitro oocyte maturation were used: (1) spontaneous gonadotropin-independent maturation and (2) follicle-stimulating hormone (FSH)-induced reversal of hypoxanthine-mediated meiotic arrest. FSH-induced, but not spontaneous germinal vesicle breakdown (GVB) was dose-dependently inhibited by both myristoylated AIP and KN-93, but not its inactive analog, KN-92. However, emission of the first polar body (PB1) was inhibited by myristoylated AIP and KN-93 in both oocyte maturation systems. Oocytes that failed to produce PB1 exhibited normal-appearing metaphase I chromosome congression and spindles indicating that CaM KII inhibitors blocked the metaphase I to anaphase I transition. Similar results were obtained when the oocytes were treated with a calmodulin antagonist, W-7, and matured spontaneously. These results suggest that CaM KII, and hence the calcium signaling pathway, is potentially involved in regulating the meiotic maturation of mouse oocytes. This kinase both participates in gonadotropin-induced resumption of meiosis, as well as promoting the metaphase I to anaphase I transition. Further evidence is therefore, provided of the critical role of calcium-dependent pathways in mammalian oocyte maturation. PMID- 11891929 TI - Metaphase arrest in amphibian oocytes: interaction between CSF and MPF sets the equilibrium. PMID- 11891927 TI - Effects of aromatase inhibition on in vitro follicle and oocyte development analyzed by early preantral mouse follicle culture. AB - In vivo studies on folliculogenesis have documented a relation among intrafollicular steroid content, follicle growth, and oocyte development. This study examined how profound changes in androgen/estrogen ratio would affect mouse in vitro follicular development. Arimidex, a potent follicular aromatase inhibitor was used for this purpose. Early preantral follicles were cultured for 12 days up to the preovulatory stage. Oocyte's meiotic maturation, spindle and chromosome configurations, in vitro fertilization and preimplantation embryo development were evaluated. Compared to controls, Arimidex reduced E2 concentration in follicle culture medium by a factor 1000, and an expected simultaneous accumulation of testosterone was measured in the conditioned medium. Arimidex treatment provoked a dose-dependent earlier differentiation of the granulosa cells as judged by an earlier antrallike cavity formation and slightly elevated basal progesterone secretion. Follicle survival exceeded 98% in all groups and all follicles responded normally to HCG/EGF addition on day 12 by cumulus mucification. By the HCG ovulatory challenge, progesterone output was reduced in Arimidex supplemented groups suggesting preovulatory luteinization. These results indicate that in vitro mouse follicles can develop normally under very low levels of estrogens and that a local androgen increase by a factor 3 is not atretogenic. Oocyte growth did not differ among culture conditions. Arimidex treatment induced a dose dependent enhancement of GVBD and polar body formation rate in response to HCG at the end of culture. Spindle and chromosome analyses demonstrated that in all groups, 90% of the oocytes which extruded a polar body had also reached the MII stage. While most of the cultured MII oocytes had a normal spindle and well aligned chromosomes, significantly less oocytes were fertilized in the groups cultured in the presence of Arimidex. Once fertilized, however, there was found to be no difference for preimplantation embryo development between controls and Arimidex treatment. These data suggest that in mice a pronounced estrogenic environment is not essential for in vitro folliculogenesis. Drastic changes in the intrafollicular steroid concentrations do not disrupt meiotic maturation nor compromise early preimplantation development, but adversely affect fertilization of in vitro grown oocytes. PMID- 11891930 TI - Introduction: evolutionary theory and the search for a unified theory of fertility. AB - Demography and evolutionary biology share common origins but have divergent emphasis on the role of theory in understanding population phenomena. A unified theory of fertility would be beneficial in explaining variation in demographic characteristics across geographic and temporal gulfs and in integrating disparate perspectives. The six papers in this thematic collection represent a nascent but vital field: human evolutionary demography. These papers examine the ways in which evolutionary theory can inform, strengthen, and focus research on topics of long-standing interest to demographers by explicitly modeling the relationship of socioecological variables to life history traits. The papers demonstrate that an understanding of human life history evolution and the use of evolutionary theory as an organizing framework can lead to a productive reassessment of five areas, which are of long-standing concern to demographers, and which conventional demographic approaches have had limited success in understanding. These are conflicts of interest between parents and children and between men and women, the allocation of resources to competing and/or alternative forms of investment in reproduction and parenting, resource flow within the household, demographic transitions and particularly the fertility transition associated with economic development, and variation in life history characteristics such as fertility and mortality across populations. Future research integrating models of trait environment correlation with models of individual information processing and decision-making will help identify areas of focus, revitalize current models, and play a leading role in the development of a unified theory of fertility applicable across societies and times. PMID- 11891931 TI - An evolutionary ecological perspective on demographic transitions: modeling multiple currencies. AB - Life history theory postulates tradeoffs of current versus future reproduction; today women face evolutionarily novel versions of these tradeoffs. Optimal age at first birth is the result of tradeoffs in fertility and mortality; ceteris paribus, early reproduction is advantageous. Yet modern women in developed nations experience relatively late first births; they appear to be trading off socioeconomic status and the paths to raised SES, education and work, against early fertility. Here, [1] using delineating parameter values drawn from data in the literature, we model these tradeoffs to determine how much socioeconomic advantage will compensate for delayed first births and lower lifetime fertility; and [2] we examine the effects of work and education on women's lifetime and age specific fertility using data from seven cohorts in the Panel Study of Income Dynamics (PSID). PMID- 11891932 TI - Demographic consequences of unpredictability in fertility outcomes. AB - Child survival is probabilistic, but the unpredictability in family formation and completed family size has been neglected in the fertility literature. In many societies, ending the family cycle with too few or too many surviving offspring entails serious social, economic, or fitness consequences. A model of risk- (or variance-) sensitive adaptive behavior that addresses long-term fertility outcomes is presented. The model shows that under conditions likely to be common, optimal, risk-sensitive reproductive strategies deviate systematically from the completed family size that would be expected if reproductive outcome is were predictable. This is termed the "variance compensation hypothesis." Variance compensation may be either positive or negative, resulting in augmented or diminished fertility. Which outcome obtained is a function of identifiable social, economic, and environmental factors. Through its effect on fertility behavior, variance compensation has a direct bearing on birth spacing and completed fertility, and thereby on problems in demography and human population biology ranging from demographic transitions to maternal depletion and child health. Risk-sensitive models will be a necessary component of a general theory of fertility. PMID- 11891935 TI - Does polygyny reduce fertility? AB - Polygyny can increase, decrease, or have no effect on fertility. Understanding how this can occur requires consideration of both the proximate determinants of fertility and the ultimate effects of polygyny as a female reproductive strategy. Several factors reduced the fertility of polygynous women in 19th century Utah, including marrying at an older age, marrying older men, and conflict between co wives. Sterility did not explain the reduced number of children in polygynous women, nor is there evidence of a "dilution effect" from sharing a husband. If women could anticipate a reduction in their own fertility, why would they still choose polygyny? Evidence suggests that they chose it because the children of polygynous men had increased fertility, high enough to offset the low fertility of polygynous women themselves. PMID- 11891934 TI - Evolutionary demography and intrahousehold time allocation: school attendance and child labor among the Okavango Delta Peoples of Botswana. AB - The ways in which resources are allocated within households and/or families, especially within the context of children's time allocation to labor and schooling, has long been a subject of concern to demographers and economists. Differential investment in children and resulting differences in activity budgets may have significant effects on children's growth and development as well as on aspects of reproduction. This study uses predictions regarding parental investment in the embodied capital of offspring generated by evolutionary theory to examine the pattern of children's time allocation to labor and schooling among the Okavango Delta Peoples of Botswana. Models incorporating individual costs and benefits of resource allocation, conflicts of interests between men and women and between parents and offspring, and the effects of family composition, subsistence ecology, and gender are developed and applied to data on time allocation, household demography, and household economy. Several findings emerged: (1) The availability of alternative productive tasks strongly affects intra- and intergenerational labor substitution. (2) The presence of similarly aged children of the same sex within the household decreases the likelihood of both boys and girls engaging in a specific productive activity and increases the likelihood of children's school attendance. (3) Birth order, the labor needs of the household, and parents' marital status all affect school attendance. These results have implications for understanding the determinants of children's time allocation to labor and schooling and consequent impacts on development, health, and welfare. PMID- 11891933 TI - Antiquity of postreproductive life: are there modern impacts on hunter-gatherer postreproductive life spans? AB - Female postreproductive life is a striking feature of human life history and there have been several recent attempts to account for its evolution. But archaeologists estimate that in the past, few individuals lived many postreproductive years. Is postreproductive life a phenotypic outcome of modern conditions, needing no evolutionary account? This article assesses effects of the modern world on hunter-gatherer adult mortality, with special reference to the Hadza. Evidence suggests that such effects are not sufficient to deny the existence of substantial life expectancy at the end of the childbearing career. Data from contemporary hunter-gatherers (Ache, !Kung, Hadza) match longevity extrapolated from regressions of lifespan on body and brain weight. Twenty or so vigorous years between the end of reproduction and the onset of significant senescence does require an explanation. PMID- 11891937 TI - Genetic admixture in three Mexican Mestizo populations based on D1S80 and HLA DQA1 loci. AB - This study compares genetic polymorphisms at the D1S80 and HLA-DQA1 loci in three Mexican Mestizo populations from three large states (Nuevo Leon, Jalisco, and the Federal District). Allele frequency distributions are relatively homogenous in the three samples; only the Federal District population shows minor differences of the HLA-DQA1 allele frequencies compared with the other two. In terms of genetic composition, these Mestizo populations show evidence of admixture with predominantly Spanish-European (50-60%) and Amerindian (37-49%) contributions; the African contribution (1-3%) is minor. Together with the observation that in Nuevo Leon, the admixture estimates based on D1S80 and HLA-DQA1, are virtually the same as those reported earlier from blood group loci, suggests that DNA markers, such as D1S80 and HLA-DQA1 are useful for examining genetic homogeneity/heterogeneity across Mestizo populations of Mexico. The inverse relationship of the proportion of gene diversity due to population differences (Gst) to within population gene diversity (Hs) is also consistent with theoretical predictions, supporting the use of these markers for population genetics studies. PMID- 11891936 TI - Evolutionary approach to below replacement fertility. AB - The large human brain, the long period of juvenile dependence, long life span, and male support of reproduction are the co-evolutionary result of the human niche based on skill-intensive techniques of resource accrual. The regulation of fertility under traditional conditions is based upon a co-evolved psychology and physiology where adjustments of investment in offspring depend upon the returns to skill and mortality hazards. When all wealth is somatic, the hormonal system controlling ovulation and implantation translates income into genetic descendants. In modern society the existence of extra-somatic wealth is a critical condition to which our evolved proximate physiological mechanisms do not respond. However, psychological mechanisms regulating parental investment in offspring quality may lead to greater and greater investment in own and offspring education, a smaller desired family size, a delay in the onset of reproduction, and a reduction in the total numbers of offspring produced. This delay in reproduction can cause many individuals to produce fewer children than desired because fecundity falls during the reproductive part of the life course. As more individuals in a society follow this pattern, more will fail to reach their desired family size. At the same time the effective use of birth control decreases the numbers of families producing more children than desired. Below replacement fertility can result. Predictions from this model were tested using data from the National Survey of Families and Households and the Albuquerque Men study. PMID- 11891938 TI - Lay perceptions of genetic risks attributable to inbreeding in Pakistan. AB - Pakistan along with many other West and South Asian countries has a very high prevalence of consanguineous, especially close cousin, marriages. Although there is substantial empirical information on offspring morbidity and mortality attributable to parental consanguinity, population-based information on how communities in general, and women in particular, perceive the health risks associated with consanguineous unions is limited. This paper considers community perceptions of health effects associated with consanguineous marriages using qualitative data from 15 focus group discussions and 294 in-depth interviews. The study was conducted in four low-income, multi-ethnic, and multi-religious communities in Karachi, the principal commercial center of Pakistan. The results show a general lack of awareness of the possible adverse health effects of consanguineous marriage. In cases where a link between consanguinity and ill health was acknowledged, it often centered on the familial origins of non communicable disorders such as diabetes and hypertension or infectious diseases such as tuberculosis. Belief in fate and the "evil eye" was widespread across all ethnic and religious groups. Many respondents did not agree with medical explanations of a genetic mode of disease inheritance, even in cases where there was an affected child in the family. The absence of a uniform manifestation of disease among all children of a couple who were identified as carriers of a specific mutation added to the confusion among participants. The study highlights the need for further quantification of risks associated with consanguinity and a need for provision of appropriate information to primary-care clinicians and also to communities. The likely impact of increasing morbidity attributable to inbreeding on the health care system in resource poor settings is also discussed. PMID- 11891939 TI - Age change of power in weight/height(p) indices used as indicators of adiposity in Japanese. AB - Data from the Statistical Report on School Health and Hygiene were used to identify the entire process of the power of height p in the weight-for-height index W/H(p). The appropriate power of height p was determined by clarifying the allometric relationship, giving an index of relative weight, which is highly correlated with weight but uncorrelated with height for age. The procedure is equivalent to fitting a straight line on a log-log scale to the tabulated weights by height in different age groups. The p values show directional changes from 5 to 17 years of age in both sexes. The optimal p value is approximately 2 in preschool children but increases gradually to over 3 at 10 years in males and 11 years in females, and then falls back to 2 after puberty. The largest p value occurs 18 months earlier in females than in males, and the pattern is the same in the present samples and American samples. Because there are remarkable changes between 12 to 14 years in females and 14 to 16 years in males, the general weight for-height indices with the precise power of height are preferable at these ages. A fixed power 3.0 is suggested for children between 3 and 12 years irrespective of sex or race. PMID- 11891940 TI - Functional status after primary surgical therapy for squamous cell carcinoma of the base of the tongue. AB - INTRODUCTION: Quality-of-life analysis is essential in determining the eventual outcome after treatment for head and neck cancer. This is particularly important when functional sequelae of treatment cause significant morbidity. The purpose of this study is to evaluate the functional status of a group of patients who had undergone primary surgical therapy for squamous cell carcinoma of the base of the tongue. METHODS: At our institution from 1979 to 1993, we identified 93 patients who had undergone resection of the base of the tongue as primary treatment for squamous cell carcinoma. Patients who required laryngectomy were excluded from this group. Forty-eight survivors were identified, and the questionnaires included the Performance Status Scale for Head and Neck Cancer Patients (PSS) and the Karnofsky Performance Status Scale (KPS). The data were reported numerically, with 0 representing the worst score and 100 representing the best score. RESULTS: Twenty-six patients completed the questionnaires. There were 19 men and 7 women. Their mean survival time was 8.6 years. Two patients had their primary tumors staged as T1, 17 patients had T2, and 7 patients had T3 disease. When evaluating the normalcy of diet, the mean score for the whole group was 73.1 (range, 20 100), the mean score for understandability of speech was 80.8 (range, 50-100), and the mean score for eating in public was 79.8 (range, 0-00). The mean KPS was 90 (range, 60-100). When comparing early (T1 and T2) with advanced (T3) disease, there were no significant differences in PSS and KPS. When comparing younger (<50 years) with older (>50 years) patients, there were no significant differences in PSS scores. Younger patients had a significantly higher KPS than older patients: mean, 97.5 vs 86.4 (p <.02). CONCLUSIONS: The long-term functional status for these patients who had undergone resection of a significant portion of the base of their tongue was good. The outcome did not seem to be related to either the stage of the lesion or the age of the patient. More studies are needed to examine the functional outcome of this patient population. PMID- 11891941 TI - Importance of the treatment package time in surgery and postoperative radiation therapy for squamous carcinoma of the head and neck. AB - BACKGROUND: To determine the effect of treatment time-related factors on outcome in patients treated with surgery and postoperative radiation therapy (RT) for locally advanced squamous cell carcinoma of head and neck (SCCHN) METHODS: A retrospective review was performed on 208 consecutive patients treated from 1992 to 1997 with surgery and postoperative RT (> or =55 Gy) for SCCHN. The treatment time factors considered were (1) interval from surgery to the start of RT; (2) RT duration; and (3) the total time from surgery to completion of RT (treatment package time). Treatment package time was dichotomized into short (< or =100 days) vs long (>100 days) categories. Other variables considered were clinical and pathologic staging, margin status, RT dose, and tumor site. Patients were also divided into intermediate- and high-risk groups on the basis of eligibility for RTOG 95-01. Univariate (logrank) and multivariate analyses were performed. RESULTS: Median follow-up for surviving patients was 24 months. Actuarial 2-year locoregional control (LRC) and survival rates were 82% and 71%, respectively. In univariate analysis, factors associated with higher locoregional failure were high-risk group (p =.011), margin status (p =.038), pathologic stage (p =.035), clinical N stage (p =.006), package time (p =.013), and RT treatment time (p =.03). Package time was also a significant predictor of survival in univariate analysis (p =.021). The other two individual time factors, tumor factors, and RT dose were not significant. Both risk status and treatment package time were significant factors in a multivariate model of LRC. CONCLUSIONS: A total treatment package time of <100 days is associated with improved tumor control and survival. Every effort should be made to keep the time from surgery to the completion of postoperative RT to <100 days. PMID- 11891943 TI - Can flow cytometrically determined DNA ploidy and S-phase fraction predict regional metastasis in squamous cell carcinoma of the oral cavity? AB - BACKGROUND: The value of flow cytometric analysis of DNA ploidy and S-phase fraction (SPF) as an indicator of regional metastasis in oral cancer is currently being debated. Intratumoral heterogeneity makes this problem complex. METHODS: Intratumoral DNA ploidy heterogeneity and intratumoral SPF variation were examined using multiple specimens from 31 surgically resected specimens taken from patients with oral cancer without preoperative therapy. Flow cytometric analysis of single biopsy specimens from 79 patients with oral cancer was also undertaken to ascertain their value as indicators of regional metastasis. RESULTS: Forty-five percent (14 of 31) of tumors showed intratumoral ploidy heterogeneity. Intratumoral SPF variation in the 31 tumors ranged from 0.2% to 6.9% (mean, 3.3%). Multivariate analysis showed that a SPF greater than 27% was the most important parameter for predicting regional metastasis. CONCLUSIONS: DNA ploidy is heterogeneous within a tumor, whereas SPF is relatively stable and can be correlated with regional metastasis in oral cancer. PMID- 11891942 TI - FDG PET studies during treatment: prediction of therapy outcome in head and neck squamous cell carcinoma. AB - BACKGROUND: Positron emission tomography (PET) provides metabolic information of tissues in vivo. The purpose of this study was to assess the value of PET with 2 [(18) F] fluoro-2-deoxy-D-glucose (FDG) in prediction of therapy outcome (tumor response, survival, and locoregional control) in locally advanced HNSCC. METHODS: Between 1993 and 1999 47 patients underwent PET before (PET(1)) and after (PET(2)) 1 to 3 weeks of radical treatment with evaluation of metabolic rate (MR) and standardized uptake value (SUV) of FDG. All patients received radiotherapy, and 10 also received neoadjuvant chemotherapy. Median follow-up time was 3.3 years. RESULTS: Low and high MR FDG at PET(2), with median value as cutoff, was associated with complete remission in 96% and 62% (p =.007), with 5-year overall survival in 72% and 35% (p =.0042) and with local control in 96% and 55% (p =.002), respectively. CONCLUSIONS: FDG PET in the early phase of treatment of HNSCC is associated with tumor response, survival, and local control. PMID- 11891944 TI - Chromosomal integration of Epstein-Barr virus genomes in nasopharyngeal carcinoma cells. AB - BACKGROUND: Little has been known about whether Epstein-Barr virus (EBV) could persist in nasopharyngeal carcinoma (NPC) cells by chromosomal integration, and no NPC cell line harboring integrated EBV has been reported. In this study, we explored this issue through isolating EBV-infected NPC cell clones generated from an in vitro infection system and examining the configuration of EBV DNA in these cells. METHODS AND RESULTS: EBV genomes were demonstrated in NPC cell clones using polymerase chain reaction and Southern hybridization. Viral nuclear antigens were also detected by use of an anticomplement immunofluorescence assay and an immunoblotting assay. Gardella gel analysis showed that two of the EBV positive cell clones, H2B4 and H2B17-7, harbored no extrachromosomal form of the viral genome. Restriction analysis of EBV genomic termini indicated that EBV DNA in these two cell clones was not circularized, and the viral genomes were integrated into chromosomes as demonstrated by fluorescence in situ hybridization. CONCLUSIONS: This is the first in vitro model of EBV persistence in NPC cells by genomic integration, which represents a unique state of virus cell interaction. Using this model, investigation into the association between EBV integration and chromosomal abnormality in tumor cells will help to reveal the underlying biologic significance. PMID- 11891945 TI - Endoglin (CD105) expression in squamous cell carcinoma of the oral cavity. AB - BACKGROUND: Increasing evidence suggests that endoglin (CD105) is a new powerful marker of neovascularization in solid malignancies. To explain the rating of CD105 expression in 51 squamous cell carcinoma of the oral cavity (SCCOC) we analyzed CD105 expression in tumor tissue and adjacent normal healthy mucosa. METHODS: Mean CD105 density was quantitated by counting the number of CD105 immunostained blood vessels. The results were compared with clinical parameters like T and N stage, grading, tumor localization, and specific characteristics of patients by means of statistical analysis. RESULTS: Endoglin expression in tumor tissue was significantly higher than in normal healthy mucosa (p <.001). With the exception of T3 (n = 2), a higher T stage was correlated with higher endoglin expression. No statistical correlation was found in the analysis of patient's age, gender, and tumor localization. CONCLUSIONS: Endoglin expression is up regulated in SCCOC compared with normal healthy oral mucosa. Endoglin may have a significant role in the development of SCCOC and might be relatively more specific than commonly used endothelial markers. PMID- 11891946 TI - Parathyroid cytology: avoiding diagnostic pitfalls. AB - BACKGROUND: Interpretation of parathyroid fine-needle aspirates (FNA) remains problematic not only because this type of specimen is rare but also because the pertinent literature is very limited. We systematically reviewed parathyroid FNAs in our files and sought to delineate additional diagnostic criteria. DESIGN: Review of all thyroid and parathyroid aspirates from January 1990 to June 1998 disclosed 12 parathyroid lesions. The final diagnoses included four parathyroid adenomas, one intrathyroidal hyperplastic parathyroid, one intrathyroidal parathyroid adenoma, one atypical parathyroid adenoma (all confirmed by histologic screening or immunocytochemistry), and five parathyroid cysts (all confirmed by immunoassay). Papanicolaou and Diff Quik-stained smears of the parathyroid FNAs were reviewed. The cytologic features were compared and contrasted with those of thyroid FNAs to establish criteria for differential diagnoses. RESULTS: The FNAs of the five parathyroid cysts yielded virtually acellular fluid with a characteristic water-clear appearance and markedly elevated levels of parathyroid hormone. The remaining seven aspirates consisted of moderately cellular smears that showed an admixture of architectural features. Common patterns included cohesive three-dimensional groups, disorganized sheets, papillary fragments, microfollicles, and a single case showing lymphoidlike smears. Although the cells were generally small and round to oval, all cases demonstrated mild to moderate anisokaryosis. The nuclei were hyperchromatic E with coarsely granular chromatin reminiscent of that of small lymphocytes. Occasional nucleoli were noted. Although the cytoplasm was usually pale blue and finely granular with ill-defined borders, two cases showed well-delineated cytoplasmic membranes. Less common findings included cytoplasmic granulation, vacuolization, and rare oxyphilic cells. Naked nuclei were noted in the background of all of the aspirates to varying degrees. Other background findings included the presence of colloidlike material, macrophages, and lymphocytes. One interesting finding that to date has not been reported is the presence of nuclear overlapping (100%) and nuclear molding (71%), which is an uncommon finding in thyroid aspirates. CONCLUSIONS: FNAs of the parathyroid can be easily confused with that of the thyroid, not only because of the clinical similarity between these two types of lesions but also because of the overlap in cytomorphologic features of the aspirated cells. Although no one single cytomorphologic feature is diagnostic, a combination of cytologic parameters noted earlier should raise the possibility of a parathyroid lesion. Aspirates of parathyroid cysts show acellular water-clear fluid with elevated parathyroid hormone measurements. PMID- 11891947 TI - Presentation, treatment, and outcome of oral cavity cancer: a National Cancer Data Base report. AB - BACKGROUND: Oral cancer has been identified as a significant public health threat. Systematic evaluation of the impact of this disease on the US population is of great importance to health care providers and policy makers. METHODS: This study used the National Cancer Data Base (NCDB) to evaluate associations between demographic and disease characteristics, treatment, and survival for patients with oral cavity cancer in the United States. Of patients diagnosed between 1985 and 1996, 58,976 were extracted from the NCDB. ANOVAs were performed on selected cross-tabulations, and relative survival was used to calculate outcome. RESULTS: Median age of patients was 64.0 years. Men made up 60.2% of patients. Pathologic diagnosis was squamous cell carcinoma (SCC) in 86.3% of cases. Younger patients had a much higher frequency of non-SCC, and this was related to survival in these patients. African-Americans (independent of income), lower income patients, and patients with higher grade disease were seen more frequently with advanced-stage SCC. Five-year relative survival for SCC cases was lower for older patients, men, and African-Americans. CONCLUSIONS: This study addressed many issues related to oral cancer that have been previously discussed in the literature. The demographic, site, stage, histologic, and survival data available for this large number of cases in the NCDB allowed an accurate characterization of the contemporary status of oral cancer in the United States. PMID- 11891948 TI - Radial clearance in resection of hypopharyngeal cancer: an independent prognostic factor. AB - BACKGROUND: The depth of infiltration of tumor is of particular relevance in hypopharyngeal cancers, because most of them are seen late, and extensive infiltration into the muscle wall and the cartilage are not uncommon. METHODS: The resected specimens of hypopharyngeal cancers were studied with whole-organ step-serial sectioning. The extent of infiltration into the thickness of the wall and the radial clearance were carefully documented. These parameters were correlated with the tumor recurrence and survival rates. RESULTS: Most patients with hypopharyngeal cancer had a minimal radial margin; the radial clearance was <1 mm in 56% of the patients. Despite such a minimal margin, the local recurrence rate was only 19% and occurred mainly in the upper and lower resection margins. Radial clearance was an independent prognostic factor for overall survival, disease-free survival, and nodal recurrence-free survival on multivariate analysis. CONCLUSION: Radial clearance is an important independent prognostic factor, and it is recommended to be included in the routine pathologic reporting of the resected specimen in hypopharyngeal cancer. PMID- 11891949 TI - Frozen section during parotid surgery revisited: efficacy of its applications and changing trend of indications. AB - BACKGROUND: Efficacy of frozen sections was assessed in terms of its various applications. The changing role of frozen sections in parotid surgery was examined. METHOD: Records of parotid operations over a 15-year period in a University Department of Head & Neck Surgery were reviewed. RESULTS: Of 241 operations, frozen sections were performed on 32. Applications of frozen sections included diagnosis, margin clearance, and checking suspicious lymph nodes and nerve invasion. The false-positive rate for malignancy was 12.5%. Margins may still be involved despite correct tissue diagnosis from sampling error. No inappropriate surgery resulted from the information obtained. With the advent of fine-needle aspiration, frozen sections were less often called for and a shift from a diagnostic role to margin checking was seen. Frozen sections picked up all unsuspected malignant tumors. CONCLUSION: Frozen sections are helpful when interpreted cautiously, but clinical assessment and fine-needle aspiration are also important components in the workup. PMID- 11891950 TI - Second primary tumors and field cancerization in oral and oropharyngeal cancer: molecular techniques provide new insights and definitions. AB - Second primary tumors (SPTs) are a significant problem in treating oral and oropharyngeal squamous cell carcinoma and have a negative impact on survival. In most studies the definition of SPT is based on the criteria of Warren and Gates, published in 1932. These criteria, however, are ill-defined and lead to confusion. Recent molecular studies have shown that a tumor can be surrounded by a mucosal field consisting of genetically altered cells. Furthermore, evidence has been provided that SPTs (defined by classical criteria) can share some or even all genetic markers with the index tumor, indicating that both tumors have arisen from a common cell clone. We propose that these secondary neoplastic lesions should not be considered SPTs, implying that the present concept of SPT needs revision. This review describes a novel classification of the secondary tumors that develop after treatment of a carcinoma in the oral cavity or oropharynx. On the basis of the molecular analysis of the tumors and the genetically altered mucosal field in between, we propose definitions for a "true SPT," a local recurrence, a "SFT" (second field tumor derived from the same genetically altered mucosal field as the primary tumor), and a metastasis. Considering the etiologic differences of these lesions, we believe that an accurate molecular definition is essential to make headway with the clinical management of oral and oropharyngeal cancer. PMID- 11891952 TI - Postauricular sebaceous carcinoma arising in association with nevus sebaceus. AB - BACKGROUND: Sebaceous carcinoma is an uncommon malignant neoplasm usually associated with the ocular adnexa. Despite the widespread anatomic distribution of sebaceous glands, extraocular sebaceous carcinoma occurs with far less frequency. METHODS: A 27-year-old man was examined with the presenting complaint of a slowly enlarging subcutaneous mass. It was associated with an overlying, irregular, verrucoid epidermal plaque. RESULTS: Histologic and electron microscopic examination of the mass revealed a malignant sebaceous neoplasm occurring in conjunction with an overlying nevus sebaceus. The dermal neoplasm contained numerous cytoplasmic and stromal hyaline globules. CONCLUSIONS: We report a case of extraocular sebaceous carcinoma arising in the postauricular region in association with a nevus sebaceus and exhibiting the unusual histologic feature of hyaline globules. PMID- 11891951 TI - Management issues in massive pediatric facial plexiform neurofibroma with neurofibromatosis type 1. AB - BACKGROUND: Plexiform neurofibroma is a relatively common but potentially devastating manifestation of neurofibromatosis type 1 (NF1). Surgical management is the mainstay of therapy, but within the head and neck region it is limited by the infiltrating nature of these tumors, inherent operative morbidity, and high rate of regrowth. METHOD: We describe a case of a 7-year-old girl with neurofibromatosis type 1 and a massive facial plexiform neurofibroma with the aim of emphasizing the treatment and timing issues involved in the management of this difficult problem. A MEDLINE search (1966 through December 2000) was carried out, and pertinent literature on the subject was reviewed. RESULT: The patient described in this case report was carefully observed for a period of 6 years from diagnosis before surgical excision of the tumor was undertaken with an uneventful recovery. CONCLUSION: Surgical management remains the mainstay of treatment for these locally invasive tumors, but functional disturbances are almost inevitable in resecting substantial tumors involving the head and neck region. The indication and timing of surgery in pediatric patients therefore needs to be carefully weighed against the physical and psychologic consequences of treatment. PMID- 11891953 TI - The usefulness of cytodiagnosis and DNA cytometry on nasopharyngeal brush smears for the diagnosis of nasopharyngeal carcinoma. AB - BACKGROUND: To assess the usefulness of combining cytodiagnosis and DNA cytometry on nasopharyngeal brush samples for the detection of nasopharyngeal carcinoma (NPC). METHODS: DNA ploidy analysis was undertaken on 66 nasopharyngeal brush samples that had been previously evaluated cytologically. RESULTS: Cytodiagnosis and ploidy analysis demonstrated a sensitivity of 66% and 55%, respectively. Both techniques had a specificity of 100% for NPC cases that were histologically confirmed. The negative predictive values for cytodiagnosis and ploidy analysis were 29% and 24%, respectively. CONCLUSIONS: The sensitivity and specificity for cytodiagnosis were not improved by the addition of DNA ploidy analysis. Furthermore, combining the two techniques did not confer any significant advantage compared with the use of cytology and DNA ploidy alone. The relatively low negative predictive values are a significant limitation, and combining cytodiagnosis and ploidy analysis on nasopharyngeal brush samples cannot be recommended for the detection of NPC. PMID- 11891954 TI - Pilot trial of concomitant chemotherapy with paclitaxel and split-course radiotherapy for very advanced squamous cell carcinoma of head and neck. AB - PURPOSE: The combination of chemotherapy and irradiation is considered the standard of care for the treatment of advanced squamous cell carcinoma of head and neck (SCCHN). Paclitaxel has shown a single-agent activity in SCCHN. Besides, this drug is a promising radiosensitizer for some human solid tumors. This is a phase II trial to evaluate the feasibility, efficacy, and toxicity of paclitaxel administered concurrently with split-course radiotherapy in advanced unresectable SCCHN. Methods and Materials Thirty-one patients with advanced SCCHN were enrolled in this trial. Radiotherapy consisted of 66 to 70 Gy delivered over 8 to 10 weeks to the primary tumor and lymphatic drainage, with a fractionation scheme of 1.8 to 2 Gy/field/d. After the initial five patients were treated, a 1-week treatment break was introduced. Paclitaxel was administered weekly in a 1-hour intravenous infusion at a projected dosage of 45 mg/m(2)/wk. RESULTS: The complete and partial response rates, based on a 4-week postradiation evaluation were 43.3% and 40%, respectively, with an overall response rate of 83.3%. Median survival was 49.4 weeks, and 1-year survival was 48%. Freedom from local progression was 65.6% at 1 year. Thirty-six percent and 20% of the patients are alive and disease free at 1 and 2 years, respectively. Grade 3/4 of acute toxicity consisted mostly of mucositis, cutaneous reaction, and weight loss. CONCLUSIONS: Paclitaxel concurrent with radiotherapy seems to be active in squamous cell carcinoma of the head and neck. In the regimen selected for this trial, toxicity was significant and led to a prolongation of treatment time. PMID- 11891955 TI - Metastatic squamous cell carcinoma of the neck from an unknown primary: management options and patterns of relapse. AB - PURPOSE: Management of squamous cell carcinoma of undetermined primary tumors in the head and neck region is controversial. Here we report the Southern California Kaiser Permanente experience with these patients. METHODS AND MATERIALS: From January 1969 through December 1994, 106 patients were eligible for this retrospective analysis. Distribution of nodal staging was as follows: 14 N1, 27 N2A, 39 N2B, 2 N2C, and 24 N3. Initial treatment included excisional biopsy alone in 12, radical neck dissection alone in 29, radiotherapy alone in 24, excisional biopsy followed by radiotherapy in 15, and radical neck dissection plus postoperative radiation in 26 patients. RESULTS: Except for two patients, all patients have had a minimum follow-up of 5 years. Overall, 57 patients (54%) have had recurrences. Only two patients (3%) who had received radiotherapy as part of their initial treatment had an appearance of a potential primary site inside the irradiated field vs 13 patients (32%) who had not received radiotherapy (p =.006). Combined modality therapy resulted in fewer neck relapses, particularly in patients with advanced neck disease. Including salvage, surgery alone as the initial treatment resulted in 81% ultimate tumor control above the clavicle for patients with N1 and N2a disease without extracapsular extension. The 5-year survival for the entire population was 53%. Radiotherapy alone resulted in poor survival in patients with advanced/unresectable neck disease. No significant difference in survival based on the initial treatment was found. The statistically significant adverse factors in determining survival included advanced nodal stage and the presence of extracapsular extension. CONCLUSIONS: Radiotherapy is very effective in reducing the rate of appearance of a potential primary site. However, in the absence of advanced neck disease (N1 and N2A without extracapsular extension), radiotherapy can be reserved for salvage. Radiotherapy alone results in poor outcomes in patients with advanced/unresectable neck disease, and incorporation of concurrent chemotherapy and cytoprotective agents should be investigated. PMID- 11891956 TI - Primary mucosal malignant melanoma of the head and neck. AB - INTRODUCTION: The relative rarity of mucosal melanomas of the head and neck (MMHN) has made analysis of treatment approaches difficult. Advances in diagnostic techniques and treatment interventions have had obvious impact on outcomes in cutaneous melanoma, but the effects on outcome in MMHN remain undefined. This study aims to assess the outcome and identify clinical and histologic prognostic indicators in a recent cohort of patients with MMHN treated at a single institution. METHODS: The clinical records of 59 patients with the diagnosis of MMHN treated at Memorial Sloan-Kettering Cancer Center (MSKCC) between 1978 and 1998 were retrospectively reviewed. Pathologic material on each of these patients was prospectively reviewed by at least two pathologists (MP, KB, or AH) for confirmation of diagnosis and assessment of histologic variables. Survival was calculated by the Kaplan-Meier method. Clinical (patient demographics, tumor characteristics, and treatment) and histologic data (tumor thickness, melanosis, melanoma in situ, vascular invasion, and multifocality) were analyzed for impact on outcome by both univariate and multivariate analyses. RESULTS: Thirty-five patients (59%) had sinonasal tumors (SNMM), whereas 24 (41%) had oral (ORMM) tumors. Forty-seven patients (79.6%) were staged as stage I, 8 (13.6%) as stage II, and 4 (6.8%) were classified as stage III. Regional lymphatic metastases at presentation were more frequent in ORMM compared with SNMM (25% vs 6%, p =.05). Surgery was used in all patients. Adjuvant radiation therapy was used more frequently in the SNMM group compared with the ORMM group (40% vs 17%, p =.04). The rates of local failure for ORMM and SNMM were 51% and 50%, nodal failure rates were 42% and 20%, and distant failure rates were 67% and 40%, respectively (p = NS). With a median follow-up of 20 months, the 5-year disease-specific survival rate was 44% (40% for ORMM vs 47% for SNMM, p = NS). Significant prognostic factors for disease-specific survival on univariate analysis included advanced clinical stage at presentation, tumor thickness greater than 5 mm, presence of vascular invasion, and development of nodal and distant metastases. On multivariate analysis, however, regional nodal failure lost significance. CONCLUSIONS: Clinical stage at presentation, tumor thickness greater than 5 mm, vascular invasion on histologic studies, and development of distant failure are the only independent predictors of outcome in MMHN. PMID- 11891957 TI - Squamous cell carcinoma of the upper aerodigestive tract: the prevalence of microscopic extracapsular spread and soft tissue deposits in the clinically N0 neck. AB - BACKGROUND: With squamous cell carcinoma of the upper aerodigestive tract the presence or absence of neck metastases is the most important prognostic factor. This makes the histopathologic assessment of neck dissections of paramount importance. With the clinically N0 neck the prevalence of microscopic extracapsular spread and soft tissue deposits has not previously been described. METHODS: We have prospectively analyzed 96 elective neck dissections in 63 patients with upper aerodigestive tract squamous cell carcinoma and clinically N0 necks to assess the prevalence of microscopic extracapsular spread and soft tissue deposits. The dissections were separated peroperatively into nodal levels; these were sectioned at 6-microm sections and stained with H & E. RESULTS: Nineteen patients (30.2%) were upstaged to pN+ve. Twelve of these had microscopic extracapsular spread, which was 19.0% of the clinically N0 necks and 63.2% of the pN+ve. Five had soft tissue deposits, which was 7.9% of the clinically N0 necks. Fourteen patients had microscopic extracapsular spread and/or soft tissue deposits, which represented 22.2% of all necks examined and 73.7% of the pN+ve necks. CONCLUSIONS: Microscopic extracapsular spread and soft tissue deposits have a high prevalence in patients with clinically N0 necks. Extracapsular spread can occur at an early stage in metastasis from upper aerodigestive tract squamous cell carcinoma. Soft tissue deposits can also occur at an early stage. Soft tissue deposits may occur by the same process as lymph node metastasis with total effacement of the lymph node or may occur by some other process such as lymphatic tumor embolization. PMID- 11891958 TI - Discrepancies in frozen section mucosal margin tissue in laryngeal squamous cell carcinoma. AB - BACKGROUND: Head and neck surgeons commonly request frozen sections. Practice patterns vary from laboratory to laboratory on how the tissue is used in performing the frozen section. Some pathologists wish to see all the material submitted by consuming it completely during frozen section, whereas others reserve some for permanent section. We wished to determine whether knowledge of margin status was initially inaccurate because of reserving tissue for permanent section. METHODS: Sixty-five laryngectomies (total and partial) with margin assessment enhanced by frozen section evaluation were studied. Forty-five laryngectomy specimens, generating 249 frozen sections in which a permanent section was prepared from tissue remaining from frozen section examination, were studied. RESULTS: Five of the 249 frozen sections contained a discrepancy between the frozen section and permanent section because of insufficient leveling of the frozen section block. These five discrepancies were called negative on frozen section, but permanent section revealed dysplasia (two cases of mild dysplasia, one case with moderate dysplasia, and one case with severe dysplasia) or carcinoma in situ (one case). Twenty laryngectomies in which the frozen section tissue was consumed at the time of frozen section generated 103 frozen sections. In eight of the frozen sections involving six cases, the diagnostic tissue was not present on one or two of the frozen section levels examined. CONCLUSIONS: We conclude that in examining margins for laryngeal squamous cell carcinoma the frozen section tissue should be completely sampled by examining several levels at the time of frozen section. This requires consuming or exhausting the frozen section tissue rather than reserving any remaining frozen tissue for a paraffin embedded permanent section. PMID- 11891959 TI - T stage as prognostic factor in irradiated localized squamous cell carcinoma of the nasal vestibule. AB - BACKGROUND: To investigate the impact of T stage according to Wang on the prognosis of irradiated nasal vestibule carcinoma. PATIENTS AND METHODS: Treatment results of 47 patients were retrospectively analyzed. Treatment consisted of external beam radiotherapy (n = 26) or interstitial radiotherapy (n = 19) or a combination of both (n = 2) for a primary, localized, squamous cell carcinoma of the nasal vestibule. Mean follow-up was 5 years and 7 months. RESULTS: T1/T2 tumors: Local control was achieved in 40 of 44 patients; surgical salvage was possible in 2 of 4 local failures. Five patients had recurrences in the neck, and four of them could be salvaged surgically. One patient had distant metastases develop. T3 tumors (n = 3): no T3 tumor could locally be cured by radiotherapy. One patient was salvaged surgically but died of regional and distant metastases. Disease-specific survival is significantly correlated with T stage according to Wang (p =.0001). Most (85%) patients were smokers, and eight of them (20%) had a second primary tumor develop in the lungs. CONCLUSIONS: The effect of radiotherapy is significantly correlated with T stage (p =.0001) and hence less successful in T3 lesions as primary treatment option. The high incidence of second primary tumors in the lung is indicative for a similar carcinogenic influence of smoking on the nasal vestibule. PMID- 11891960 TI - High dose rate brachytherapy for early stage oral tongue cancer. AB - BACKGROUND: High dose rate (HDR) interstitial brachytherapy of the oral tongue is a new treatment modality. Our study evaluates the outcomes of patients with early stage oral tongue cancer as treated by HDR interstitial implant. METHODS: We reviewed the records of 19 patients who were seen between 1994 and 2000 with carcinoma of the oral tongue and whose primary tumors were treated solely with interstitial implant using HDR remote afterloading technique. Ten patients had T1 N0 disease, and the remaining 9 had T2 N0 disease. Elective neck treatment was withheld for 12 patients. The remaining seven patients had ipsilateral elective neck dissection. The male-female ratio was 1:0.9, and the median age was 60 years (range, 32-81 years). The median follow-up time was 43 months (range, 6-78 months). The afterloading catheters were positioned by the submandibular approach with the assistance of a template set. Fifteen patients had single planar implants, and the remaining four had double planar implants. The median number of catheters inserted was 5 (range, 4-9). The median dose given was 55 Gy in 10 fractions over 6 days. The minimal interfraction interval was 7 hours for the first 7 patients and was extended to 8 hours for the other 12. Mandibular shields were inserted before treatment. RESULTS: The mucositis lasted for 6 to 20 weeks (median, 9 weeks). One patient had local failure, and the 4-year local failure free survival rate was 94.7%. Three of the 12 patients without elective neck treatment had ipsilateral regional failure develop. They were salvaged by neck node dissection and regionally remained in control. One patient with multiple nodal metastases and extracapsular spread had biopsy-proven liver metastases and died 6 months after implant. One of the seven patients who were treated with elective neck dissection had multiple nodal metastases and extracapsular spread. She was treated with postoperative radiotherapy to the neck. She died 30 months after implant with evidence of regional and distant failure. One patient treated with double planar implant had grade II necrosis of the soft tissue and bone develop. The necrosis resolved with conservative treatment. Another four patients had small area of soft tissue deficit of the tongue attributed to aggressive debulking or biopsy before brachytherapy. CONCLUSIONS: Our experience in treating early stage tongue cancer with HDR remote afterloading technique is encouraging, because it gives a local control rate of 94.7% at 4 years with acceptable morbidity. Further studies are eagerly awaited to delineate the optimum schedule for this new treatment modality. PMID- 11891961 TI - Wait-and-see policy for the N0 neck in early-stage oral and oropharyngeal squamous cell carcinoma using ultrasonography-guided cytology: is there a role for identification of the sentinel node? AB - BACKGROUND: Management of the N0 neck in patients with head and neck squamous cell carcinoma (SCC) remains controversial. We describe the outcome of patients who underwent transoral tumor excision and a wait-and-see policy for the neck staged N0 by ultrasonography-guided cytology (USgFNAC). Because selection of lymph nodes for USgFNAC is currently based on size criteria, we investigated the additional value of sentinel node (SN) identification. METHODS: The outcome of 161 patients with T1-T2 oral/oropharyngeal SCC was determined. In a subgroup of 39 patients the SN was identified and aspirated in addition. RESULTS: SN identification and aspiration was possible in 38 of 39 patients but without decreasing the false-negative rate of USgFNAC. During follow-up (12-99 months) 34 of 161 (21%) patients developed lymph node metastases. After therapeutic neck dissection and postoperative radiotherapy, 27 of 34 (79%) could be salvaged (88% regional control). CONCLUSIONS: Wait-and-see seems justified in case of negative USgFNAC. Strict follow-up with USgFNAC is required. SN identification and aspiration is feasible but did not improve lymph node selection. PMID- 11891962 TI - Genomic instability measurement in the diagnosis of thyroid neoplasms. AB - BACKGROUND: Clinically palpable thyroid nodules are present in approximately 10% of the population, although only 5% to 7% of these nodules harbor malignancy. Fine-needle aspiration has become one of the central tools in the diagnostic armamentarium of the surgeon/endocrinologist. There is, however, up to a 30% indeterminate diagnostic rate associated with this technique, resulting in unnecessary surgical interventions for patients harboring benign disease. A second issue of clinical importance is the unreliability of predicting outcomes based either on histologic findings alone or in combination with clinical staging. To address these diagnostic and clinical shortcomings, we have used measurement of genomic instability as a diagnostic and prognostic indicator for thyroid neoplasms. METHODS: Genomic instability of thyroid tissue samples was determined by inter-(simple sequence repeat) PCR, microsatellite instability analysis, and fluorescence in situ hybridization (FISH) on thyroid neoplasms from 22 patients. RESULTS: Inter-(simple sequence repeat) PCR detected genomic instability with an index range 0% to 1.9% (mean, 0.56%) in patients with benign disease, whereas in patients with malignant histologic findings the values ranged from 0% to 6.6% (mean, 2.9%). This difference between benign and malignant values was statistically significant (p =.004). There was no demonstrable microsatellite instability or loss of heterozygosity for six markers examined in this group. Losses of chromosomes 17 and X in benign disease and gains of chromosomes 7, 12, 17, and X in Hurthle cell carcinoma were observed, although not at a significant rate. CONCLUSIONS: Genomic instability as measured by inter-(simple sequence repeat) PCR was significantly higher for malignant diseases compared with benign thyroid tissues, but no such association was seen with aneuploidy or microsatellite instability. PMID- 11891963 TI - Soft tissue sarcoma. PMID- 11891964 TI - Synchronous benign and malignant salivary gland tumors in ipsilateral glands: a report of two cases and a review of literature. AB - BACKGROUND: Ipsilateral salivary gland tumors of different histologic types are rare and make up less than 0.3% of all salivary gland neoplasms. Only nine cases of synchronous benign and malignant ipsilateral parotid gland tumors have been described in the literature. METHODS: Two additional cases of synchronous benign and malignant neoplasms in the parotid gland are reported and discussed with a review of literature. RESULTS: Our first case describes a pleomorphic adenoma and a salivary duct carcinoma, an entity not previously reported in the literature. The second case documents the most common benign and malignant ipsilateral parotid gland neoplasm reported in this case series, a Warthin's tumor and a mucoepidermoid carcinoma. CONCLUSIONS: Synchronous salivary gland tumors exhibiting both benign and malignant components are uncommonly observed, with only nine cases published to date. We describe two additional cases of a synchronous benign and malignant ipsilateral parotid gland tumor. PMID- 11891965 TI - Eosinophilic angiocentric fibrosis of the sinonasal tract: report on the clinicopathologic features of a case and review of the literature. AB - BACKGROUND: Eosinophilic angiocentric fibrosis (EAF) is a rare fibroinflammatory lesion of the sinonasal tract that occurs mainly in young to middle-aged female patients. Only two previous cases affecting male patients have been reported, and its etiopathogenesis remains unknown. The authors report on the third case of the entity in a male patient and review the 12 previously reported cases. CASE REPORT: A 52-year-old male patient was initially seen with a 15 years history of allergic rhinitis, progressive nasal obstruction, and left-sided hearing loss. All laboratory tests were unremarkable, except the nasal discharge eosinophil count that showed a conspicuous eosinophilia. The video-assisted-nasofibroscopic examination and CT scans disclosed a thickened deviated nasal septum with a subjacent infiltrative lesion. The histologic analysis of the nasal septum showed a variable mixed inflammatory cellular infiltration mainly composed of eosinophils, plasma cells, and histiocytes with a perivascular distribution; in other areas, an angiocentric fibrosing lesion with a peculiar perivascular onion skin pattern was observed. The patient had a partial resection of the lesion with symptomatic control. CONCLUSIONS: The presence of rhinitis and nasal eosinophilia in our case associated with the clinical aspects of the previously reported cases further support an allergic cause for EAF. PMID- 11891966 TI - Is there a BOLD response of the visual cortex on stimulation of the vision related acupoint GB 37? AB - PURPOSE: To determine whether or not acupuncture of guangming (GB 37) produces a significant response of the visual cortex detectable by means of functional magnetic resonance imaging (fMRI). MATERIALS AND METHODS: This study investigates the activation of the visual cortex elicited by a soft and an intensified stimulation of GB 37, an acupoint documented to influence vision-related disorders. Three different paradigms were carried out to detect any possible modulation of the Blood Oxygenation Level Dependent (BOLD)-response in the visual cortex to visual stimulation through acupuncture. RESULTS: The percentage signal changes in the visual stimulation cycles did not significantly differ before vs. during acupuncture. CONCLUSION: Whereas no BOLD-response correlating with acupuncture was detected in the visual cortex, BOLD-signal-changes in response to needle twisting were detected in different cortical areas. Further studies are necessary to clarify whether these clusters correlate to inevitable somatosensory stimulation accompanying acupuncture or represent an acupuncture-specific response. PMID- 11891967 TI - Dynamic contrast-enhanced magnetic resonance imaging as a surrogate marker of tumor response to anti-angiogenic therapy in a xenograft model of glioblastoma multiforme. AB - PURPOSE: To evaluate the effects of a neutralizing anti-vascular endothelial growth factor (anti-VEGF) antibody on tumor microvascular permeability, a proposed indicator of angiogenesis, and tumor growth in a rodent malignant glioma model. MATERIALS AND METHODS: A dynamic contrast-enhanced magnetic resonance imaging (MRI) technique, permitting noninvasive in vivo and in situ assessment of potential therapeutic effects, was used to measure tumor microvascular characteristics and volumes. U-87, a cell line derived from a human glioblastoma multiforme, was implanted orthotopically into brains of athymic homozygous nude rats. RESULTS: Treatment with the monoclonal antibody A4.6.1, specific for VEGF, significantly inhibited tumor microvascular permeability (6.1 +/- 3.6 mL min( 1)100 cc(-1)), compared to the control, saline-treated tumors (28.6 +/- 8.6 mL min(-1)100 cc(-1)), and significantly suppressed tumor growth (P <.05). CONCLUSION: Findings demonstrate that tumor vascular permeability and tumor growth can be inhibited by neutralization of endogenous VEGF and suggest that angiogenesis with the maintenance of endothelial hyperpermeability requires the presence of VEGF within the tissue microenvironment. Changes in tumor vessel permeability and tumor volumes as measured by contrast-enhanced MRI provide an assay that could prove useful for clinical monitoring of anti-angiogenic therapies in brain tumors. PMID- 11891968 TI - Neurostimulation systems for deep brain stimulation: in vitro evaluation of magnetic resonance imaging-related heating at 1.5 tesla. AB - PURPOSE: To assess magnetic resonance imaging (MRI)-related heating for a neurostimulation system (Activa Tremor Control System, Medtronic, Minneapolis, MN) used for chronic deep brain stimulation (DBS). MATERIALS AND METHODS: Different configurations were evaluated for bilateral neurostimulators (Soletra Model 7426), extensions, and leads to assess worst-case and clinically relevant positioning scenarios. In vitro testing was performed using a 1.5-T/64-MHz MR system and a gel-filled phantom designed to approximate the head and upper torso of a human subject. MRI was conducted using the transmit/receive body and transmit/receive head radio frequency (RF) coils. Various levels of RF energy were applied with the transmit/receive body (whole-body averaged specific absorption rate (SAR); range, 0.98-3.90 W/kg) and transmit/receive head (whole body averaged SAR; range, 0.07-0.24 W/kg) coils. A fluoroptic thermometry system was used to record temperatures at multiple locations before (1 minute) and during (15 minutes) MRI. RESULTS: Using the body RF coil, the highest temperature changes ranged from 2.5 degrees-25.3 degrees C. Using the head RF coil, the highest temperature changes ranged from 2.3 degrees-7.1 degrees C.Thus, these findings indicated that substantial heating occurs under certain conditions, while others produce relatively minor, physiologically inconsequential temperature increases. CONCLUSION: The temperature increases were dependent on the type of RF coil, level of SAR used, and how the lead wires were positioned. Notably, the use of clinically relevant positioning techniques for the neurostimulation system and low SARs commonly used for imaging the brain generated little heating. Based on this information, MR safety guidelines are provided. These observations are restricted to the tested neurostimulation system. PMID- 11891969 TI - Intracranial oscillations of cerebrospinal fluid and blood flows: analysis with magnetic resonance imaging. AB - PURPOSE: To detect oscillations of the cerebrospinal fluid (CSF) flow related to the heartbeat and frequencies lower than 0.6 Hz and to compare these oscillations of CSF and blood flow in cerebral vessels by using echo planar imaging in real time mode. The existence of such waves has been well known but has not yet been shown by MRI. MATERIALS AND METHODS: In a slice perpendicular to the aqueduct, CSF flow as well as CBF, could be determined in sagittal sinus, basilar artery, and capillary vessels. After Fourier analysis, four frequency bands were assigned. RESULTS: In the very high-frequency (heart rate) range, the integrals under the CSF curves were more closely related to arterial CBF than to changes in the sinus. Also, in the high-frequency (respiration rate), low-frequency (0.05 0.15 Hz), and very-low-frequency (0.008-0.05 Hz) ranges, the integrals under the CSF curves corresponded with arterial and capillary CBF. CONCLUSION: Slow and fast oscillations in CSF flow are detectable in healthy persons with a proportional allotment to arterial and capillary CBF. PMID- 11891970 TI - The reproducibility and sensitivity of brain tissue volume measurements derived from an SPM-based segmentation methodology. AB - PURPOSE: To investigate the reproducibility of SPM99-based whole brain, gray matter, and white matter volume measurements with and without image inhomogeneity correction, subsequently exploring age and gender effects on absolute and fractional (proportional to intra-cranial) volumes. MATERIALS AND METHODS: Twenty seven control subjects (aged 23.2 to 55.2 years) had three-dimensional fast spoiled gradient recall scans. Ten subjects were scanned about 197 days later. RESULTS: Coefficients of variation (CV) for absolute and fractional volumes determined from images processed with inhomogeneity correction ranged from 1.2% to 0.5%. Inhomogeneity correction reduced the CV for all measures except gray matter fractional (GMF) volumes. Significantly lower white matter absolute (WM) and fractional (WMF) volumes, and higher GMF were found in females compared with males, overlying age-related reductions (in decreasing order of significance) in brain parenchymal fraction, GMF, WMF, brain parenchymal, and gray matter volumes. CONCLUSION: SPM99 segmentations are sufficiently reproducible to detect age and gender effects in limited cohorts. PMID- 11891971 TI - MRI in lung transplant recipients using hyperpolarized 3He: comparison with CT. AB - PURPOSE: To elucidate the ability of 3He-MRI to detect ventilation defects in lung transplant recipients, 3He-MRI was compared to CT for concordance. MATERIALS AND METHODS: We examined 14 lung recipients using 3He-MRI on a 1.5 T MR scanner. CT served as a reference method. Up to four representative ventilation defects were defined for each lung on 3He-MRI and compared to corresponding areas on CT. RESULTS: Altogether, 59 representative ventilation defects were defined on 3He MRI. Plausible CT correlates were found for 29 ventilation defects; less plausible CT correlates were found for eight defects. In 22 defects (37%) no corresponding CT changes were detected. CT demonstrated correlates for ventilation defects seen on 3He-MRI in only 63% of the cases. CONCLUSION: 3He-MRI yields a clear increase in the number of detected ventilation defects compared to CT. This may have an important impact on the early detection of bronchiolitis obliterans in lung transplant recipients. PMID- 11891972 TI - Magnetic resonance imaging in patients with pancreatitis: evaluation of signal intensity and enhancement changes. AB - PURPOSE: To evaluate the utility of unenhanced and enhanced T1-weighted fat suppressed (T1-FS) magnetic resonance imaging (MRI) in detecting pancreatitis. MATERIALS AND METHODS: 1.5-T MRI was performed in 25 patients with acute and 23 patients with chronic pancreatitis and in 20 control subjects without known pancreatic disease. T1-FS spin-echo and contrast-enhanced arterial-predominant (DYN1) and portal-predominant (DYN2) fast multiplanar spoiled gradient-echo (FMPSPGR) sequences were evaluated. These three sets of images were evaluated both subjectively for decreased or heterogeneous signal intensity (rating scale, 0-3) and objectively (region of interest (ROI)) in the head, body, and tail of the pancreas, in each patient. RESULTS: Good correlation between subjective assessment and objective data was demonstrated. The T1-FS sequence showed an abnormality with greater frequency (T1-FS > DYN1, 81/144 scores; T1-FS = DYN1, 63/144 scores; T1-FS < DYN1, 0/144 scores) and magnitude (average subjective score, 2.48 vs. 1.74; P < 0.0003) than that of the contrast-enhanced FMPSPGR (decreased or heterogeneous enhancement). The overall sensitivity and specificity of MRI was 92% and 50%, respectively. On the basis of signal intensity and enhancement, MRI was not able to differentiate acute from chronic pancreatitis. CONCLUSION: MRI was highly sensitive for disease detection, particularly using the T1-FS sequence, but using the sequences described, was not able to differentiate acute from chronic pancreatitis. PMID- 11891973 TI - MR imaging of diffuse adenomyosis changes after GnRH analog therapy. AB - PURPOSE: To evaluate uterine changes on MRI before and after GnRH analog (GnRHa) treatment in diffuse adenomyosis. MATERIALS AND METHODS: Thirty-one patients with MRI features suggestive of diffuse adenomyosis received GnRHa for 6 months. Diffuse adenomyosis was sub-classified as: symmetric (symmetric/ entire widening of the junctional zone [JZ]) and asymmetric (asymmetric/ partial widening of JZ). Pre- and post-high signal intensity (SI) foci and JZ width, and post-demarcated change (interface of adenomyosis with the myometrium became more discrete with a concomitant decrease in JZ width) were analyzed. RESULTS: Before therapy, 15 of 18 asymmetric contained high SI foci compared to none of symmetric. After therapy, JZ width decreased (P < 0.0001). Eight asymmetric and none of symmetric showed demarcated change with resolved high SI foci. CONCLUSION: Our results suggest the use of GnRHa is associated with a decrease of JZ width in adenomyosis. Asymmetric adenomyosis with high SI foci appears to be the most sensitive to hormonal therapy. PMID- 11891974 TI - Combined time-resolved and high-spatial-resolution 3D MRA using an extended adaptive acquisition. AB - PURPOSE: To combine the benefits of time-resolved dynamic imaging and single elliptical centric acquisitions in a reasonable scan time. MATERIALS AND METHODS: A time series of images with moderate spatial resolution was acquired using the 3D Time-Resolved Imaging of Contrast KineticS (3D TRICKS) technique with elliptical centric encoding during contrast arrival. Following venous opacification, a complete large centrically encoded k-space volume was acquired. The high-spatial-frequency portions of this volume were combined with a 3D TRICKS time frame to form a high-resolution image. An additional single image is formed by suppressing background and signal averaging all acquired data, including post venous low-spatial-frequency data. For this image, 2D temporal correlation analysis is used to suppress low-spatial-frequency vein contributions. Arrival time and spatial correlations are used to suppress background. RESULTS: The 3D TRICKS time frame may be selected to ensure a combined high-resolution image that has optimal central k-space sampling for any vascular region. The single image formed by signal averaging all acquired data has increased contrast-to-noise (CNR) and signal-to-noise (SNR) ratios. CONCLUSION: The advantages of time resolved and high-spatial-resolution imaging were combined using an extended dual phase acquisition. Some SNR and CNR gain was achieved by signal averaging. This process is facilitated by background and vein suppression. PMID- 11891975 TI - Diffusion-weighted MRI in the characterization of soft-tissue tumors. AB - PURPOSE: To explore the potential of perfusion-corrected diffusion-weighted magnetic resonance imaging (MRI) in characterizing soft-tissue tumors. METHODS AND MATERIALS: Diffusion-weighted MRI was performed in 23 histologically proven soft-tissue masses using a diffusion-weighted spin-echo sequence with diffusion gradient strengths yielding five b-values (0-701 seconds/mm(2)). True diffusion coefficients and perfusion fractions were estimated and compared with apparent diffusion coefficients (ADCs). RESULTS: ADC values of all tumors, subcutaneous fat, and muscle were significantly higher than true diffusion coefficients, indicating a contribution of perfusion to the ADC. True diffusion coefficients of malignant tumors (1.08 x 10(-3) mm(2)/second) were significantly lower than those of benign masses (1.71 x 10(-3) mm(2)/second), whereas ADC values between these groups were not significantly different. CONCLUSION: Perfusion-corrected diffusion-weighted MRI has potential in differentiating benign from malignant soft-tissue masses. PMID- 11891976 TI - Blood perfusion of vertebral lesions evaluated with gadolinium-enhanced dynamic MRI: in comparison with compression fracture and metastasis. AB - PURPOSE: To investigate blood perfusion of vertebral lesions using dynamic Gd DTPA-enhanced MRI. MATERIALS AND METHODS: Dynamic MR studies were performed for cases of acute compression fracture, chronic compression fracture, metastatic vertebral lesions with or without compression fracture. A total of 42 patients (71 vertebral segments) were included. Five types of time-intensity curves (TICs) were defined as: nearly no enhancement (type A), slow enhancement (type B), a rapid contrast wash-in followed by an equilibrium phase (type C), a rapid contrast wash-in followed by early wash-out (type D), and a rapid contrast wash in with a second slower-rising slope (type E). RESULTS: Metastatic vertebral lesions with or without fracture had a higher peak enhancement percentage and steeper enhancement slope than those of chronic compression fracture, but had no difference as compared to those of acute compression fracture. The type D curve had high positive predictive value for metastatic group (100%), and the type E curve had high positive predictive value for benign compression fracture (85.7%). CONCLUSION: Type D and E curves are valuable in the differentiation of benign and malignant vertebral lesions. PMID- 11891977 TI - Effect of IL-1beta-induced macromolecular depletion on residual quadrupolar interaction in articular cartilage. AB - PURPOSE: Sodium multiple-quantum filtered (MQF) NMR spectroscopy may potentially be used to measure proteoglycan (PG) depletion in cartilage caused by osteoarthritis (OA). The purpose of this work was to quantify the effect of interleukin-1 (IL-1beta)-induced macromolecule depletion on the residual quadrupolar interaction (RQI) of sodium in bovine cartilage plugs. MATERIALS AND METHODS: Fifteen 8-mm-diameter cartilage plug specimens were cored from the articular surface of fresh bovine patellae. All plugs were kept in culture media and nine of the plugs were subjected to interleukin-1 (IL-1beta)-induced degeneration of cartilage for 4, 6, and 7 days. Sodium NMR spectra were obtained from each sample with a 1-cm-diameter solenoid coil in a 2T whole-body magnet interfaced to a custom-built spectrometer. We employed a previously described theoretical model to analyze triple-quantum filtered (TQF) and double-quantum filtered magic angle (DQFMA) spectra obtained from normal cartilage and cartilage treated with IL-1beta. The model assumes a static Gaussian distribution of the RQI frequency, omega(Q), in the sample. TQF and DQFMA spectra from each sample were fitted with the appropriate signal expressions to determine sigma (the root mean square (RMS) omegaQ), T2f, and T2s. An inversion-recovery sequence was used to determine T1 of each plug. A spectrophotometric assay was used to determine the amount of PG depleted from each plug. Histology was performed to visualize the PG loss in cartilage plugs. We defined sigma as the measure of changes in macroscopic order in the tissue. RESULTS: Simulated spectra from the theoretical model were in excellent agreement with the experimental data. We were able to determine the relaxation times as well as sigma of each specimen from their corresponding fits. T2f ranged between 2.26-3.50 msec, decreasing with increased PG loss. Over the range of PG depletion investigated, T2s increased from 12.3 msec to 14.9 msec, and T1 increased from 16 msec to 21 msec, while sigma decreased from 180 Hz to 120 Hz. The order of macromolecules in the cartilage tissue decreased substantially with PG loss. Histology sections clearly showed qualitative visualization of the PG loss in cartilage following treatment with IL 1beta. CONCLUSION: We demonstrated that IL-beta-induced macromolecule depletion in cartilage not only changes the relaxation characteristics of sodium but also changes RQI of the tissue. Using MQF sodium spectroscopy we quantified the changes in sigma and showed that loss of macromolecules reduces the degree of order in the tissue. PMID- 11891978 TI - MRI of the pelvic ring joints postpartum: normal and pathological findings. AB - PURPOSE: To assess the joints of the pelvic ring postpartum and to discern normal postpartum findings and pathologic lesions using MRI. MATERIALS AND METHODS: MR images were obtained in six women with severe pelvic ring pain after delivery, in 13 women after uncomplicated vaginal delivery, and in 11 healthy, nulliparous non pregnant volunteers. Distances of the pubic gap, signal intensities of the pubic cartilage, and signal changes of the pelvic ring bones were determined and evaluated. RESULTS: Both postpartum groups had significantly larger distances of the interpubic gap compared to the nulliparous group (P = 0.0002). The mean signal intensity of cartilage of the symphysis pubis was significantly different on the T1-weighted and T2-weighted MR images in postpartum women compared to nulliparous women (P = 0.001), indicating a higher water content of the pubic cartilage. 13 of all 19 postpartum women had bruises of parasymphyseal pubic bones. One pubic symphysis rupture and one sacral stress fracture were detected in two symptomatic women. CONCLUSION: MR imaging is a useful adjunct to clinical examination to identify patients with lesions of the pelvic ring postpartum. MRI of the pelvic ring of asymptomatic postpartum women can demonstrate signal changes of the pubic cartilage and small bruises of the pubic bones. PMID- 11891980 TI - Effects of iodinated contrast and field strength on gadolinium enhancement: implications for direct MR arthrography. AB - PURPOSE: To optimize direct magnetic resonance (MR) arthrography by determining the effect of dilution of gadolinium in iodinated contrast, saline, or albumin on T1-weighted, T2-weighted, and gradient-recalled echo (GRE) images, and the effect of scanner field strength. MATERIALS AND METHODS: Gadopentetate dimeglumine was diluted into normal saline, albumin, or iodinated contrast (0.625 mmol/liter to 40 mmol/liter). Samples were scanned at 1.5T and 0.2T. Signal intensity was measured using T1-weighted spin-echo (SE), T2-weighted SE, and two- and three dimensional GRE (20 degrees-75 degrees flip angle) sequences. Graphical analysis of signal intensity vs. gadolinium concentration was performed. RESULTS: Albumin had no effect on gadolinium contrast. Dilution of gadolinium in iodinated contrast decreased signal intensity on all sequences compared to samples of identical concentration diluted in saline at both 1.5T and 0.2T: with a 2 mmol/liter gadolinium solution at 1.5T, signal was decreased by 26.1% on T1 weighted images, 31.7% on GRE20 images, and 28.9% on GRE45 images, and the T2 value decreased by 71.1%; at 0.2T, signal was decreased by 23.5% on T1-weighted images. On all sequences, the peak signal shifted to the left (lower gadolinium concentration) when diluted in iodinated contrast. Peak signal was also seen at different gadolinium concentrations on different sequences and field strength: at 1.5T, peak in saline/iodine was 2.5/0.625 mmol/liter on T1-weighted images, and 2.5/1.25 mmol/liter on GRE20 and GRE45 sequences. At 0.2T, peak in saline/iodine was 0.625-2.5/1.25 mmol/liter on T1-weighted images, 0.625-2.5/1.25 on GRE45 images, 2.5-10.0/1.25-5.0 mmol/liter on GRE65 images, and 1.25-5.0/0.625-1.25 mmol/liter on GRE75 images. CONCLUSION: Dilution of gadolinium in iodinated contrast results in decreased signal on T1-weighted, T2-weighted, and GRE images compared to dilution in saline or albuminfor both 1.5-T and 0.2-T scanners; if gadolinium is diluted in iodinated contrast for MR arthrography, a lower concentration should be used because the peak is shifted to the left. The use of iodinated contrast should be minimized, as it may diminish enhancement and lower the sensitivity and specificity of MR arthrography. Optimal gadolinium concentration for MR arthrography is dependent on scanner field strength and a broader range of gadolinium concentration can be used to provide maximal signal at low field strength. PMID- 11891979 TI - Arterioportal shunts mimicking hepatic tumors with hyperintensity on T2-weighted MR images. AB - We report and assess the imaging findings in a patient with multiple arterioportal shunting mimicking multiple hypervascular tumors that showed multiple areas of hyperintensity with gadolinium-enhanced gradient-recalled-echo images obtained in the hepatic arterial phase and corresponding areas of slight hyperintensity with T2-weighted images. PMID- 11891981 TI - Evaluation of a dedicated dual phased-array surface coil using a black-blood FSE sequence for high resolution MRI of the carotid vessel wall. AB - PURPOSE: To investigate the ability of magnetic resonance imaging (MRI) to visualize the carotid vessel wall using a phased-array coil and a black-blood (BB) fast spin-echo (FSE) sequence. MATERIALS AND METHODS: The phased-array coil was compared with a three-inch coil. Images from volunteers were evaluated for artifacts, wall layers, and wall signal intensity. Signal intensity and homogeneity of atherosclerosis were assessed. Lumen diameter and vessel area were measured. RESULTS: Comparison between the phased-array coil and the three-inch coil showed a 100% increase in signal-to-noise ratio. BB-FSE imaging resulted in good delineation between blood and vessel wall. Most volunteers had a two-layered vessel wall with a hyperintense inner layer. MRI showed both homogeneous hyperintense and heterogeneous plaques, which consisted of a main hyperintense part with hypointense spots and/or intermediate regions. MRI lumen and area measurements were performed easily. CONCLUSION: High resolution MRI of carotid atherosclerosis is feasible with a phased-array coil and a BB-FSE sequence. PMID- 11891982 TI - Wherefore heart thou? Embryonic origins of cardiogenic mesoderm. AB - The developing heart in avian embryos has been examined extensively over the past several decades using classic embryologic and, more recently, molecular and genetic approaches. Still, conflicting reports arise as to the location and regulation of early heart progenitors in the embryo. In addition, a new source of cardiomyocytes has been identified recently that contributes to the outflow tract after the heart initially forms. The focus of this review is the examination of the tissue interactions, signaling molecules, and gene regulatory mechanisms that, together, control heart formation from primary and secondary heart forming fields of the embryo. Early studies of the induction and regulation of the secondary heart field indicate that at least some of the events of primary cardiomyogenesis are recapitulated when the conotruncal myocardium is recruited into the heart. The consideration of classic embryologic studies of the heart forming fields in terms of modern molecular and genetic tools provides reinforcing evidence for the location of cardiac progenitors in the embryo. The accurate definition of early cardiac regulatory events provides a necessary foundation for the generation of new therapeutic sources of cardiomyocytes. PMID- 11891983 TI - Differential expression of the seven-pass transmembrane cadherin genes Celsr1-3 and distribution of the Celsr2 protein during mouse development. AB - Drosophila Flamingo (Fmi) is an evolutionally conserved seven-pass transmembrane receptor of the cadherin superfamily. Fmi plays multiple roles in patterning neuronal processes and epithelial planar cell polarity. To explore the in vivo roles of Fmi homologs in mammals, we previously cloned one of the mouse homologs, mouse flamingo1/Celsr2. Here, we report the results of our study of its embryonic and postnatal expression patterns together with those of two other paralogs, Celsr1 and Celsr3. Celsr1-3 expression was initiated broadly in the nervous system at early developmental stages, and each paralog showed characteristic expression patterns in the developing CNS. These genes were also expressed in several other organs, including the cochlea, where hair cells develop planar polarity, the kidney, and the whisker. The Celsr2 protein was distributed at intercellular boundaries in the whisker and on processes of neuronal cells such as hippocampal pyramidal cells, Purkinje cells, and olfactory neurons. Celsr2 is mapped to a distal region of the mouse chromosome 3. We discussed possible functions of seven-pass transmembrane cadherins in mouse development. PMID- 11891984 TI - Region- and stage-specific effects of FGFs and BMPs in chick mandibular morphogenesis. AB - The mandibular processes are specified as at least two independent functional regions: two large lateral regions where morphogenesis is dependent on fibroblast growth factor (FGF)-8 signaling, and a small medial region where morphogenesis is independent of FGF-8 signaling. To gain insight into signaling pathways that may be involved in morphogenesis of the medial region, we have examined the roles of pathways regulated by FGFs and bone morphogenetic proteins (BMPs) in morphogenesis of the medial and lateral regions of the developing chick mandible. Our results show that, unlike in the lateral region, the proliferation and growth of the mesenchyme in the medial region is dependent on signals derived from the overlying epithelium. We also show that medial and lateral mandibular mesenchyme respond differently to exogenous FGFs and BMPs. FGF-2 and FGF-4 can mimic many of the effects of mandibular epithelium from the medial region, including supporting the expression of Msx genes, outgrowth of the mandibular processes and elongation of Meckel's cartilage. On the other hand, laterally placed FGF beads did not induce ectopic expression of Msx genes and did not affect the growth of the mandibular processes. These functional studies, together with our tissue distribution studies, suggest that FGF-mediated signaling (other than FGF-8), through interactions with FGF receptor-2 and downstream target genes including Msx genes, is part of the signaling pathway that mediates the growth-promoting interactions in the medial region of the developing mandible. Our observations also suggest that BMPs play multiple stage- and region-specific roles in mandibular morphogenesis. In this study, we show that exogenous BMP-7 applied to the lateral region at early stages of development (stage 20) caused apoptosis, ectopic expression of Msx genes, and inhibited outgrowth of the mandibular processes and the formation of Meckel's cartilage. Our additional experiments suggest that the differences between the effects of BMP-7 on lateral mandibular mesenchyme at stage 20 and previously reported results at stage 23 (Wang et al., [1999] Dev. Dyn. 216:320-335) are related to differences in stages of differentiation in that BMP-7 promotes apoptosis in undifferentiated lateral mandibular mesenchyme, whereas it promotes chondrogenesis at later stages of development. We also showed that, unlike mandibular epithelium and medially placed FGF beads, medially placed BMP-7 did not support outgrowth of the isolated mesenchyme and at stage 20 induced the formation of a duplicated rod of cartilage extending from the body of Meckel's cartilage. These observations suggest that BMPs do not play essential roles in growth-promoting interactions in the medial region of the developing mandible. However, BMP-mediated signaling is a part of the signaling pathways regulating chondrogenesis of the mandibular mesenchyme. PMID- 11891986 TI - Immediate early genes krox-24 and krox-20 are rapidly up-regulated after wounding in the embryonic and adult mouse. AB - Embryos show a remarkable capacity for perfect repair after injury. Wounding of embryonic skin triggers rapid activation of epithelial sweeping and mesenchymal contraction tissue movements that bear striking analogy to several naturally occurring morphogenetic tissue movements, but very little is known about the early molecular signals that might initiate such movements. Here, we describe the rapid and transient up-regulation of two immediate early genes, krox-24 and krox 20, after wounding of the embryonic mouse. Furthermore, we demonstrate that these signals are conserved, but of longer duration, in the neonate and adult wound situation. To further test the roles of these transcription factors in vivo, we performed wound healing studies on embryos lacking either Krox-24 or 20. Despite the dramatic up-regulation of these genes in response to injury, our studies reveal that neither of them on their own is essential for repair. PMID- 11891985 TI - Analysis of two distinct retinoic acid response elements in the homeobox gene Hoxb1 in transgenic mice. AB - Expression of vertebrate Hox genes is regulated by retinoids such as retinoic acid (RA) in cell culture and in early embryonic development. Retinoic acid response elements (RAREs) have been identified in Hox gene regulatory regions, suggesting that endogenous retinoids may be involved in the direct control of Hox gene patterning functions. Previously, two RAREs located 3' of the murine Hoxb1 gene, a DR(2) RARE and a DR(5) RARE, have been shown to regulate Hoxb1 mRNA expression in the neural epithelium and the foregut region, respectively; the foregut develops into the esophagus, liver, pancreas, lungs, and stomach. We have now examined the functional roles of these two types of 3' RAREs in regulating Hoxb1 expression at different stages of gestation, from embryonic day 7.5 to 13.5, in transgenic mice carrying specific RARE mutations. We demonstrate that the DR(5) RARE is required for the regulation of Hoxb-1 transgene region-specific expression in the gut and extraembryonic tissues, as well as for the RA-induced anteriorization of Hoxb-1 transgene expression in the gut. In contrast, expression of the Hoxb1 transgene in the neural epithelium requires only the DR(2) RARE. By in situ hybridization, we have identified a new site of Hoxb1 expression in the developing forelimbs at approximately day 12.5, and we show that, in transgenic embryos, expression in the forelimb buds requires that either the DR(2) or the DR(5) RARE is functional. Attainment of a high level of Hoxb1 transgene expression in other regions, such as in rhombomere 4 (r4) and in the somites, requires that both the DR(2) and DR(5) RAREs are functional. In addition, our transgenic data indicate that the Hoxb1 gene is expressed in other tissues such as the hernia gut, genital eminence, and lung. Our analysis shows that endogenous retinoids act through individual DR(2) and DR(5) RAREs to regulate Hoxb1 expression in different regions of the embryo and that functional redundancy between these DR(2) and DR(5) RAREs does not exist with respect to neural epithelium and the gut Hoxb1 expression. PMID- 11891988 TI - Formation of supernumerary muscle spindles at the expense of Golgi tendon organs in ER81-deficient mice. AB - ER81, a member of the ETS family of transcription factors, is essential for the formation of connections between sensory and motor neurons in the spinal cord. Mice lacking Er81 genes exhibit reduced monosynaptic sensory-motoneuron connectivity in response to muscle nerve stimulation. Proximal muscle nerve stimulation elicits fewer monosynaptic potentials than stimulation of distal nerves in hindlimbs, a deficit that is paralleled by a paucity of muscle spindles in proximal muscles (Arber et al., 2000). We examined whether a presence of spindles innervated by afferents in distal muscles correlated with the increased preservation of monosynaptic sensory-motor potentials in distal muscle nerves. Not only were spindles and Ia afferents present, but also they were supernumerary in distal muscles such as the soleus, medial gastrocnemius, and extensor hallucis longus. Concomitantly, a deficiency of Golgi tendon organs (GTOs) and Ib afferents was observed in distal muscles, as if supernumerary spindles formed at the expense of tendon organs in the absence of Er81. Thus, ER81 may be involved in mechanisms that regulate acquisition of the Ia and Ib phenotypes by subsets of proprioceptive muscle afferents. Segmental differences in muscle spindle and GTO dependence on ER81 suggest that more than one ETS transcription factor may participate in the regulation of limb proprioceptive system assembly in the mouse. PMID- 11891987 TI - Expression of the boc gene during murine embryogenesis. AB - BOC is a receptor-like protein that, with the related factor CDO, belongs to a newly recognized subfamily within the immunoglobulin superfamily of cell-surface molecules. BOC and CDO form complexes that positively regulate myogenesis in vitro. To gain a better understanding of the role of boc during vertebrate embryogenesis and whether it could cooperate with cdo in vivo, we carried out an extensive in situ hybridization analysis of boc expression in mouse embryos from E7.0 to E17.5. Our results show that boc is widely expressed during murine embryogenesis, in a spatially and temporally restricted pattern. Overall, the highest levels of boc expression are detected between E10.5 and E15.5, with the strongest signals observed in the developing musculoskeletal and central nervous systems. At late stages of development, boc expression becomes more restricted and is limited primarily to regions harboring proliferating cells, undifferentiated cells, or both. This expression pattern is strikingly similar to that described for cdo (Mulieri et al., 2000). The overlapping expression patterns of cdo and boc in vivo, combined with the promotion of myogenesis by CDO/BOC complexes in cultured cells, strongly suggests that these proteins play a role together in the determination, differentiation, or both, of numerous cell types during vertebrate embryogenesis. PMID- 11891989 TI - Matrix metalloproteinases mediate the dismantling of mesenchymal structures in the tadpole tail during thyroid hormone-induced tail resorption. AB - It has been suggested that a family of tissue remodelling enzymes called matrix metalloproteinases (MMPs) play a causal role in the process of tail resorption during thyroid hormone-induced metamorphosis of the anuran tadpole; however, this hypothesis has never been directly substantiated. We cloned two new Xenopus MMPs, gelatinase A (MMP-2) and MT3-MMP (MMP-16), and the MMP inhibitor TIMP-2. These clones were used along with several others to perform a comprehensive expression study. We show that all MMPs and TIMP-2 are dramatically induced in the resorbing tail during spontaneous metamorphosis and are spatially coexpressed, primarily in the remodelling mesenchymal tissues. By Northern blotting, we show that all the examined MMPs/TIMP-2 are also induced by treatment of organ-cultured tails with thyroid hormone (T(3)). Using the organ culture model, we provide the first direct evidence that MMPs are required for T(3)-induced tail resorption by showing that a synthetic inhibitor of MMP activity/expression can specifically retard the resorption process. By gelatin zymography, we also show T(3) induction of a fifth MMP, preliminarily identified as gelatinase B (GelB; MMP-9). Moreover, T(3) not only induces MMP/TIMP expression but also MMP activation, and we provide evidence that TIMP-2 participates in the latter process. These findings suggest that MMPs and TIMPs act in concert to effect the dismantling of mesenchymal structures during T(3)-induced metamorphic tadpole tail resorption. PMID- 11891990 TI - RP59, a marker for osteoblast recruitment, is also detected in primitive mesenchymal cells, erythroid cells, and megakaryocytes. AB - We recently described a novel protein in bone marrow of rats, RP59, as a marker for cells with the capacity to differentiate into osteoblasts. In this work, its expression pattern was further investigated to learn about the origin and biological relevance of RP59 expressing marrow cells. As revealed by in situ hybridization and by immunohistochemistry of yolk sac embryos, RP59 was found in the cells of the primitive ectoderm and primitive streak as well as in blood islands and extraembryonal mesoderm. Later, RP59 occurred in fetal liver cells and in circulating blood. From the time around birth, it was found in bone marrow and spleen cells. In addition, in vitro-formed blood vessels contained RP59 positive cells in the lumen. Endothelial cells and the vast majority of cells outside the blood vessels were not labeled. Concerning more mature hematopoietic cell types, RP59 was observed in megakaryocytes and nucleated erythroblasts, but absent from lymphoid cells. In conclusion, RP59 was induced in early mesoderm. It was maintained in the erythroid and megakaryotic lineages and, as earlier described, in young osteoblasts. PMID- 11891991 TI - Transient expression of TIP60 protein during early chick heart development. AB - Screening of an embryonic chick cDNA library revealed a gene product termed chick TIP60 (cTIP60) due to its homology with human TIP60, a founding member of the "MYST" family of proteins that possess functional motifs, including chromo, zinc finger, and histone acetyltransferase domains. cTIP60 expression was assessed during early chick embryogenesis, at the RNA level by using reverse transcriptase polymerase chain reaction (RT-PCR) and at the protein level by using Western blotting and immunohistochemistry. RT-PCR indicated that cTIP60 transcripts in whole embryos are present as early as Hamburger-Hamilton (HH) stage 5, diminishing after HH10. Western blotting of total embryonic protein revealed that cTIP60 was present in uniform quantities between HH3 and HH25. By contrast, Western blotting of protein from isolated hearts revealed that cTIP60 protein was strongly expressed at the earliest stages of heart development (HH11-13), diminishing thereafter. This finding was corroborated by immunohistochemistry, which revealed that cTIP60 protein was selectively expressed at high levels in the myocardium between HH 10-14. Considered in the context of its functional domains, these findings suggest that cTIP60 modulates transcriptional processes which regulate terminal cell differentiation, proliferation, or both, during early myocardial development. PMID- 11891993 TI - Prepulse inhibition deficits in patients with panic disorder. AB - Prepulse inhibition (PPI) is an operational measure of sensorimotor gating that is reduced in several neuropsychiatric disorders that are characterized by deficits in inhibition or gating of intrusive or irrelevant stimuli. Clinically, panic disorder (PD) patients have been described as having difficulties in inhibition of responding to sensory and cognitive events. Because such difficulties may be due to failures in early stages of information processing, we examined PPI in patients with PD. Acoustic startle reactivity, habituation, and PPI (30-, 60-, 120-, 240-, and 2,000-ms interstimulus intervals) were assessed in patients with panic disorder (m/f = 10, 10) and age- and gender-matched healthy controls (m/f = 11, 10). PD patients were assessed with structured clinical interview for DSM-IV criteria with benzodiazepine treatment as an exclusion criterion. Panic disorder patients exhibited normal startle reactivity, reduced habituation, and significantly reduced PPI in the 30-, 60-, and 240-ms prepulse conditions. Within the PD group, the patients with high trait and state anxiety exhibited less PPI than patients with low trait and state anxiety. Furthermore, in PD patients, decreased PPI correlated significantly with high trait but not state anxiety. These data indicate that early stages of sensory information processing are abnormal in patients with PD. These observed deficits in PPI could reflect a more generalized difficulty in suppressing or gating information in panic disorder. The correlation between high trait anxiety and deficient PPI supports the hypothesis that sensorimotor gating abnormalities are an enduring feature of subjects with PD. PMID- 11891994 TI - Measuring anxiety: parent-child reporting differences in clinical samples. AB - This study examines parent-child reporting differences for childhood anxiety in normal controls (n = 16) and in children with diagnosed anxiety disorders (ANX; n = 15), attention-deficit hyperactivity disorder (ADHD; n = 15), and comorbid ANX and ADHD (n = 16). Correspondence between child reports of anxiety on two self report inventories and diagnosis based on structured parent interview was assessed for all four groups. Parent-child agreement did not appear to be measurement dependent but did differ by diagnostic group, with poorer agreement for clinical groups. Though needing replication, these findings suggest that it is inadvisable to rely exclusively on self-report measures when assessing childhood anxiety, especially in clinical populations. Such measures can be useful in monitoring clinical progress, however, provided parent and child reports are examined separately. PMID- 11891995 TI - Childhood trauma in obsessive-compulsive disorder, trichotillomania, and controls. AB - There is relatively little data on the link between childhood trauma and obsessive-compulsive/putative obsessive-compulsive spectrum disorders. The revised Childhood Trauma Questionnaire (CTQ), which assesses physical, emotional, and sexual abuse as well as physical and emotional neglect, was administered to female patients with obsessive-compulsive disorder (OCD; n = 74; age: 36.1 plus minus 16.3), TTM (n = 36; age: 31.8 plus minus 12.3), and a group of normal controls (n = 31; age: 21.5 plus minus 1.0). The findings showed a significantly greater severity of childhood trauma in general, and emotional neglect specifically, in the patient groups compared to the controls. Although various factors may play a role in the etiology of both OCD and trichotillomania (TTM), this study is consistent with some evidence from previous studies suggesting that childhood trauma may play a role in the development of these disorders. PMID- 11891996 TI - Generalizability and correlates of clinically derived panic subtypes in the population. AB - To determine the generalizability and population-based correlates of clinically derived panic attack subtypes among adults in the community. Data were drawn from the National Comorbidity Survey (n = 8,098), a representative sample of adults of age 18-54 years in the United States. Sociodemographic characteristics, psychiatric comorbidity, and panic symptomotology associated with three clinically derived panic subtypes (early-onset, agoraphobia, and dyspnea) were compared using two-way ANOVA and multivariate logistic regression analyses. Among those with panic attacks in the community, 51.2% had early-onset, 32.6% had agoraphobia, and 64.4% had dysthymia. Significant differences in sociodemographic characteristics, psychiatric comorbidity, and panic symptomotology emerged between the three groups. Early-onset panic was associated with significantly increased likelihood of bipolar disorder and substance dependence but was not distinguished from the other two subtypes by panic symptoms. Panic attack with agoraphobia was associated with significantly higher odds of several comorbid anxiety disorders, and panic with dyspnea was more common among married females with less education and high levels of comorbid alcohol and depressive disorders. These data suggest that clinically derived panic subtypes are generalizable and may be associated with several unique sociodemographic and psychiatric correlates in the general population. Observed differences between these subtypes may influence results from clinical samples. PMID- 11891997 TI - Davidson Trauma Scale (DTS): normative scores in the general population and effect sizes in placebo-controlled SSRI trials. AB - The Davidson Trauma Scale (DTS) was developed as a self-rating for use in diagnosing and measuring symptom severity and treatment outcome in post-traumatic stress disorder (PTSD); 630 subjects were identified by random digit dialing and evaluated for a history of trauma. Prevalence rates of PTSD and subthreshold PTSD with impairment were 2.2 and 4.1%, respectively. In this general population sample, 438 subjects endorsed at least one trauma, and four groups were generated: A) threshold PTSD (n = 13), B) subthreshold PTSD with impairment (n = 26), C) subthreshold PTSD without impairment (n = 78), and D) no PTSD (n = 321). Mean (SD) DTS score in the entire population was 11.0 +/- 18.1. Differences were found in four of the five pairwise between-group contrasts. In a second sample of 447 clinical trial participants from three SSRI vs. placebo studies, we assessed treatment effect size according to different measures. In all three clinical trials, effect size with the DTS was equal to, or better than, those found for the Impact of Event Scale (IES), Clinician Administered PTSD Scale (CAPS), and Structured Interview for PTSD (SIP). These results further affirm the utility of the DTS as a self-rating measure of PTSD symptom severity and in evaluating treatment response. PMID- 11891998 TI - Ethnic differences in worry in a nonclinical population. AB - The present study examined ethnic differences in worry in a college student population. No differences were found between Caucasians, African Americans, and Asian Americans in pathological worry as measured by the Penn State Worry Questionnaire (PSWQ) or in the frequency with which they met self-report criteria for generalized anxiety disorder on the Generalized Anxiety Disorder Questionnaire for DSM-IV (GAD-Q-IV). Groups differed in Worry Domains Questionnaire (WDQ) total scores and on all WDQ domain subscales except for the Financial domain. Within ethnic groups, Caucasians and African Americans experienced variations in intensity of worry across the specific domains, but Asian Americans did not. These results suggest that ethnic groups may differ from each other in the degree to which they worry and in the breadth of their concerns. Further examination of ethnic differences and worry (and anxiety more generally) is suggested. PMID- 11891999 TI - Trichotillomania and skin-picking: a phenomenological comparison. AB - Although trichotillomania and pathological skin-picking are both characterized by repetitive self-injurious stereotypic behaviors, the former is classified as an impulse control disorder, while the latter is not given a specific diagnostic category in the Diagnostic and Statistical Manual of Mental Disorders (4th edition) [APA, 1994]. There are, however, few empirical data on phenomenological similarities and differences between these disorders. Patients with trichotillomania and pathological skin-picking were compared in terms of several demographic (age, gender), clinical (comorbid axis I and II disorders), and personality variables. Trichotillomania and pathological skin-picking were very similar in demographics, psychiatric comorbidity, and personality dimensions. Dissociative symptoms may be more common in trichotillomania than in pathological skin-picking. These data support the concept of phenomenological overlap between trichotillomania and pathological skin-picking. Future work to assess the implications of overlap for clinical evaluation and intervention in the two conditions may be useful. PMID- 11892000 TI - Treatment of late-onset OCD following basal ganglia infarct. AB - Current consensus on the treatment of obsessive-compulsive disorder (OCD) includes cognitive behavior therapy (CBT) in the form of exposure and response prevention (ERP). However, the generalizability of these methods to elderly populations remains largely undocumented. This clinical case study examines the effectiveness of medications and intensive, inpatient ERP in an elderly patient with onset of OCD following basal ganglia infarcts. There was a dramatic reduction from baseline to follow-up in both obsessions and compulsions with Yale Brown Obsessive-Compulsive Scale [YBOCS; Goodman et al., 1989] total scores decreasing by over 20 points. These gains were maintained up to 1 year post treatment. Age-specific issues and the application of standard therapeutic methods to elderly clients are discussed. PMID- 11892001 TI - Localization of implanted EEG electrodes in a virtual-reality environment. AB - In the planning of epilepsy surgery procedures, intracranial electrodes are implanted in a significant fraction of the patients. Accurate localization of the individual electrode contacts with respect to the brain cortex is imperative. Because the manual tracking of an EEG electrode in a CT scan in a slice-by-slice fashion is cumbersome and subjective, the goal of this study was to develop an easier and more accurate way to localize implanted EEG electrodes. In this paper, we present our solution in the form of a virtual-reality environment with interactive tools to assist the clinician with EEG localization. With the help of a high-quality and fast volume renderer, a view is created of the inside of the patient's skull to obtain an overview of the electrodes in relation to the cortical structures. Depth, grid, and reed electrodes are characterized semi interactively using different methods. For depth electrodes, the contacts (which are not visible in the CT scan) are derived by measuring off the theoretical distance between the contact and the end of the electrode from the central axis produced by a three-dimensional (3D) line tracker. For grid electrodes, the contacts are visible in a CT, so the 3D view is merely used to find the contacts and to resolve the overlap of grids with other grids, tail wires, or bone ridges. For reed electrodes, the contacts, which are again not visible in this case, are calculated from a line model fitted to the positions of lead markers. After letting the user place artificial spheres on the lead markers and wire, a B spline is fitted to the spheres' centers to estimate the positions of the contacts. The approach was evaluated by applying it to CT scans of seven patients. It appeared that the method is generally applicable (even crossing electrodes or electrodes with gaps were correctly characterized), and that the display via 3D views and slices is so good that manual placement of spheres performed as well as semi-automatic placement. From computer experiments, it appeared that the final localization error in the position of EEG contacts could be estimated to lie in the order of the dimensions of one voxel. PMID- 11892002 TI - A surface-matching technique for robot-assisted registration. AB - Successful implementation of robot-assisted surgery (RAS) requires coherent integration of spatial image data with sensing and actuating devices, each having its own coordinate system. Hence, accurate estimation of the geometric relationships between relevant reference frames, known as registration, is a crucial procedure in all RAS applications. The purpose of this paper is to present a new registration scheme, along with the results of an experimental evaluation of a robot-assisted registration method for RAS applications in orthopedics. The accuracy of the proposed registration is appropriate for specified orthopedic surgical applications such as Total Knee Replacement. The registration method is based on a surface-matching algorithm that does not require marker implants, thereby reducing surgical invasiveness. Points on the bone surface are sampled by the robot, which in turn directs the surgical tool. This technique eliminates additional coordinate transformations to an external device (such as a digitizer), resulting in increased surgical accuracy. The registration technique was tested on an RSPR six-degrees-of-freedom parallel robot specifically designed for medical applications. A six-axis force sensor attached to the robot's moving platform enables fast and accurate acquisition of positions and surface normal directions at sampled points. Sampling with a robot probe was shown to be accurate, fast, and easy to perform. The whole procedure takes about 2 min, with the robot performing most of the registration procedures, leaving the surgeon's hands free. Robotic registration was shown to provide a flawless link between preoperative planning and robotic assistance during surgery. PMID- 11892003 TI - Precision of ACL tunnel placement using traditional and robotic techniques. AB - The objective of this study was to examine the precision of ACL tunnel placement using: (1) CASPAR (orto MAQUET GmbH Co. KG)--an active robotic system, and (2) four orthopedic surgeons with various levels of experience (between 100 and 3,500 ACL reconstructions). The robotic system and each surgeon drilled tunnels for ACL reconstruction in 10 plastic knees (total n = 50) that included a reference cube in the medial aspect of the proximal tibia and distal femur. For the robotic system, the placement of each tunnel was planned preoperatively using custom software and CT data for each femur and tibia. The robotic system then drilled the tunnels in the femur and tibia based on the preoperative plan. For the surgeons, tunnel placement was accomplished using their preferred technique, which was based on the one-incision arthroscopic technique. The distribution of intra-articular points on the tibia was contained within a sphere of radius 2.0 mm (robot system), 2.1 mm (Fellow 1), 2.4 mm (Fellow 2), 3.4 mm (Experienced Surgeon 1), or 2.0 mm (Experienced Surgeon 2). On the femur, no significant differences in the distribution of intra-articular points could be demonstrated between the robotic system (2.1 mm), Fellow 1 (4.5 mm), Fellow 2 (4.1 mm), Experienced Surgeon 1 (2.3 mm), and Experienced Surgeon 2 (3.0 mm). The direction of the tunnels drilled in the femur and tibia was different with the robotic and traditional techniques. However, the robotic system had the most consistent tunnel directions, while the surgeons' tunnels were more dispersed. Variation in surgeon precision of tunnel placement for ACL reconstruction is greater on the femur than the tibia, and this can be correlated with experience. Our data also suggest that the robotic system has the same precision as the most experienced surgeons. PMID- 11892004 TI - Accuracy in tunnel placement for ACL reconstruction. Comparison of traditional arthroscopic and computer-assisted navigation techniques. AB - The purpose of this randomized, prospective study was to compare accuracy in tunnel placement as performed with a traditional arthroscopic anterior cruciate ligament (ACL) reconstruction technique and with KneeNavTM ACL, a computer assisted surgical navigation technique. Two surgeons experienced in ACL reconstruction, but inexperienced in computer-assisted surgical navigation technique, each randomly used traditional arthroscopic guides or KneeNavTM ACL to drill a tunnel in twenty identical foam knees. Placement of the resulting tibial and femoral tunnels was measured with a computer-assisted digitizing method and compared to traditional biplanar radiographs. Statistical analysis with Student's t-test was used to compare the distance from the ideal tunnel placement to the femoral and tibial tunnels. Accuracy of tunnel placement with KneeNavTM ACL was significantly better than that obtained with the traditional arthroscopic technique. Distances from the ideal tunnel placement to the femoral and tibial tunnels were 4.2 +/- 1.8 mm (mean +/- SD) and 4.9 +/- 2.3 mm, respectively, for the traditional arthroscopic technique, and 2.7 +/- 1.9 mm (femur) and 3.4 +/- 2.3 mm (tibia) for KneeNavTM ACL. These differences were statistically different. Tunnel placement for ACL reconstruction with KneeNavTM ACL, an image-based, computer-assisted surgical navigation device with a simple and intuitive interface, was more accurate than with the traditional arthroscopic technique. PMID- 11892005 TI - Comparison of fit and fill between anatomic stem and straight tapered stem using virtual implantation on the ORTHODOC workstation. AB - The objective of this article was to determine the influence of stem design on fit and fill using the preoperative planning workstation of the ROBODOC system. Anatomic ABG and straight Osteolock femoral components were virtually implanted into 50 femora (25 from patients with developmental dysplasia of the hip (DDH), 25 morphologically normal) on the workstation display. Fit and fill, and length of the proximal posterolateral femoral cortex removed by milling (LPFCR), were measured on the cross-sectional images. Lateral curvature (alpha angle) and anteversion of the femur were evaluated. The ABG components showed significantly better fit than the Osteolock components at the levels proximal to the lesser trochanter. The Osteolock components showed significantly greater LPFCR than the ABG components, especially in the patients with DDH. The patients with DDH showed significantly greater alpha angle and femoral anteversion than those with morphologically normal femora. With the Osteolock components, the alpha angle correlated significantly with femoral anteversion and LPFCR. Use of an anatomic proximal body of the stem helped to improve the proximal canal fit. Greater LPFCR was required when a straight stem was implanted in patients with a relatively high alpha angle. PMID- 11892006 TI - The use of open MRI in otorhinolaryngology: initial experience. AB - Intraoperative imaging in head and neck surgery is a useful tool in many situations. In addition to being helpful for intraoperative orientation, real time imaging enables visualization of the progress of surgery and the completeness of tumor resection. Regions in the head and neck to which access is difficult, and which therefore have a high incidence of morbidity and risk for the patient, can be approached more easily and safely in an open MRI than in a conventional way. Interventions in the open MRI (Signa SP, 0.5 Tesla) were performed with nonmagnetic instruments and an MR-safe microscope. For intraoperative navigation, the integrated FlashPoint system is helpful, because it allows targeting of the tumor by a calculated virtual line. T1W spin-echo, T2W fast spin-echo, and 3D T1W gradient-echo sequences were used for high-resolution imaging. Real-time imaging is achieved by fast multiplanar spoiled gradient-echo sequences or T2 single-shot fast spin-echo sequences. From 1996 to the present, we biopsied 17 petroclival tumors, performed paranasal sinus surgery in five cases, biopsied two neck masses, and inserted tubes for brachytherapy in 12 cases. No complications were observed. In all surgical procedures, a good resolution was obtained with MRI, especially for soft-tissue structures. The tumor could be targeted exactly, and all specimens revealed the relevant histology. In paranasal sinus surgery, however, the success rate was lower because it was difficult to distinguish blood from pathologic tissue. The insertion of tubes for brachytherapy was successful in all cases. It was possible to apply the tubes parallel to each other, 1 cm apart. Relevant biopsies could be taken of both neck masses. The indications for the use of open MRI in otorhinolaryngology are biopsies of tumors in regions that are difficult to approach, such as the petrous apex and petroclival region, the parapharyngeal space, and the orbit. Furthermore, the open MRI can be useful in paranasal sinus surgery, in the evaluation of tissue resection, and in the detection of the anatomy of delicate structures such as the internal carotid artery, the skull base, and the orbit. In addition, active navigation in the open MRI is possible with the integrated FlashPoint system. The advantage over conventional navigation systems lies in the possibility of real-time imaging, which allows detection of tissue changes occurring during the procedure. PMID- 11892007 TI - Sperm entry position provides a surface marker for the first cleavage plane of the mouse zygote. AB - The sperm entry position (SEP) of the mouse egg, labelled by placing a bead at the fertilisation cone, tends to be associated with the first cleavage plane (Piotrowska and Zernicka-Goetz: Nature 409:517-521, 2001). Nevertheless, in up to one-fourth of embryos the cleavage furrow did not pass close to the bead, and following the division the bead marked the cleavage plane in only 60% of cases. This raised the question of whether such variability arose from the labelling itself or had a biological basis. The zona pellucida was not responsible for this effect because similar results were obtained in its presence or absence. However, this variability could be attributable to the large size of the fertilisation cone relative to the SEP. Therefore, we have developed a means of fluorescently labelling sperm that can record the exact site of its penetration when the label transfers to the egg surface. This approach indicates that the SEP marks the first cleavage in the great majority (88%) of embryos. In conclusion, direct sperm labelling shows precisely the correlation between the SEP and the first cleavage, although there is natural variability in this process. PMID- 11892008 TI - A Cre/loxP-deleter transgenic line in mouse strain 129S1/SvImJ. AB - A Cre recombinase expression cassette was inserted into the X-linked Hprt locus by gene targeting in a mouse embryonic stem (ES) cell line isogenic to strain 129S1/SvImJ (129S1), then the transgene was introduced into 129S1 mice through ES cell chimeras. When females hemizygous for this transgene were mated to males carrying a neomycin selection cassette flanked by loxP sites, the cassette was always excised regardless of Cre inheritance and without detectable mosaicism. The usefulness of this "Cre-deleter" transgenic line is in its efficiency and defined genetic status in terms of mouse strain and location of the transgene. PMID- 11892009 TI - Genetic manipulation of mouse embryonic stem cells by mutant lambda integrase. AB - Mutant lambda integrases catalyze site-specific recombination reactions inside mammalian cells. Here we demonstrate that the integrase system can be used to eliminate resistance marker genes from the genome of mouse embryonic stem cells. So-called integrative and excisive recombination pathways led to the precise deletion of the neomycin gene, which was inserted together with a flanking pair of directly repeated recombination sites into the ROSA26 locus by standard targeting techniques. The excision of the resistance gene led to the expression of enhanced green fluorescence protein, which served as a means to sort out cells that had undergone site-specific recombination. Southern analysis and DNA sequencing confirmed that strand exchange reactions had occurred in the genome as expected. Hence, the integrase system may be used in conjunction with other site specific recombinases as a tool in genome manipulation protocols. PMID- 11892010 TI - Cre/loxP recombination-activated neuronal markers in mouse neocortex and hippocampus. AB - A new strategy for visualizing neuronal cell morphology of mouse brain based on Cre/loxP recombination-activated gene expression is described. A "reporter" transgenic line was generated which expressed a fusion gene encoding a dendrite targeted green fluorescent protein (MAP2-GFP) upon deletion of a transcription/translation STOP (transcription and translation stop signal) cassette. Cre transgenic "deleter" lines were established that activated reporter gene expression at various frequencies in pyramidal neurons in the forebrain. A deleter line was identified which activated a MAP2-GFP reporter gene at very low frequency (less than 0.1% of pyramidal neurons) and allowed the visualization of dendritic structures of individual neocortical and hippocampal pyramidal neurons. In addition, vertical "columns" of pyramidal neurons in the neocortex were labeled in these mice. In a second deleter line, a MAP2-GFP reporter gene was selectively activated in pyramidal neurons of the CA-1 subregion of the hippocampus in young mice. With its combinatorial property, this binary recombination-activated neuronal marker system should facilitate the study of detailed morphology, connectivity, and plasticity of defined classes of live neurons in vitro and in vivo. PMID- 11892011 TI - Quaking is essential for blood vessel development. AB - For nearly 40 years functional studies of the mouse quaking gene (qkI) have focused on its role in the postnatal central nervous system during myelination. However, the homozygous lethality of a number of ENU-induced alleles reveals that quaking has a critical role in embryonic development prior to the start of myelination. In this article, we show that quaking has a previously unsuspected and essential role in blood vessel development. Interestingly, we found that quaking, a nonsecreted protein, is expressed in the yolk sac endoderm, adjacent to the mesodermal site of developing blood islands, where the differentiation of blood and endothelial cells first occurs. Antibodies against PE-CAM-1, TIE-2 and SM-alpha-actin reveal that embryos homozygous for the qk(k2) allele have defective yolk sac vascular remodeling and abnormal vessels in the embryo proper at midgestation, coinciding with the timing of embryonic death. However, these mutants exhibit normal expression of Nkx2.5 and alpha-sarcomeric actin, indicating that cardiac muscle differentiation was normal. Further, they had normal embryonic heart rates in culture, suggesting that cardiac function was not compromised at this stage of embryonic development. Together, these results suggest that quaking plays an essential role in vascular development and that the blood vessel defects are the cause of embryonic death. PMID- 11892012 TI - The aryl hydrocarbon receptor (AhR) and its nuclear translocator (Arnt) are dispensable for normal mammary gland development but are required for fertility. AB - The aryl hydrocarbon receptor (AhR) and its nuclear translocator (Arnt) are transcription factors that play a role in the detection of and adaptation to environmental signals. AhR-null mice are viable but show impaired lactation. Deletion of the Arnt gene from the mouse genome results in embryonic lethality. To determine the role of Arnt in mammary development and function, we inactivated the Arnt gene in mammary epithelium using Cre-loxP recombination. Inactivation of the Arnt gene during pregnancy did not disrupt alveolar development or the ability of dams to nurse their litters. In contrast, dams in which the Arnt gene had been inactivated during puberty and in ovaries were subfertile, exhibited retarded mammary development, and impaired mammary function. To distinguish defects autonomous to mammary epithelium from indirect effects controlled by ovarian hormones, we transplanted Arnt-null and AhR-null mammary epithelium into wild-type mice and evaluated development after one pregnancy. Normal mammary structures were observed in the absence of Arnt and AhR, demonstrating that neither transcription factor is necessary for mammary development. PMID- 11892013 TI - Expression pattern of the Brachyury gene in the arrow worm paraspadella gotoi (chaetognatha). AB - Arrow worms (the phylum Chaetognatha), which are among the major marine planktonic animals, are direct developers and exhibit features characteristic of both deuterostomes and protostomes. In particular, the embryonic development of arrow worms appears to be of the deuterostome type. Brachyury functions critically in the formation of the notochord in chordates, whereas the gene is expressed in both the blastopore and stomodeum invagination regions in embryos of hemichordates and echinoderms. Here we analyzed the expression of Brachyury (Pg Bra) in the arrow worm Paraspadella gotoi and showed that Pg-Bra is expressed in the blastopore region and the stomodeum region in the embryo and then around the mouth opening region at the time of hatching. The expression of Pg-Bra in the embryo resembles that of Brachyury in embryos of hemichordates and echinoderms, whereas that in the mouth opening region in the hatchling appears to be novel. PMID- 11892014 TI - Bethesda 2001: Science, technology, and democracy join forces for patient care. PMID- 11892015 TI - Estrogen and progesterone hormone receptor status in breast carcinoma: comparison of immunocytochemistry and immunohistochemistry. AB - We evaluated the correlation between histologic and cytologic specimens in the determination of estrogen receptor (ER) and progesterone receptor (PR) status in breast carcinoma and investigated the causes of clinically significant discrepancies. We analyzed 70 immunoassays for ER and 60 for PR from 71 patients with breast carcinoma. Concordance between cytology and histology was 89% for ER and 63% for PR using scores from pathology reports. Concordance between cytology and histology was 98% for ER and 91% for PR using consensus scores (obtained after reevaluation by the team of pathologists). Thirty of 130 (23%) tests had clinically relevant discrepancies, 53% of which were caused by wrong interpretation of cytologic findings, 10% by wrong interpretation of histologic findings, 17% by sampling error and 20% were not available for reevaluation. Wrong interpretation of the results for ER and PR status in cytology was a far more frequent cause of clinically relevant discrepancies than sampling errors. The use of strict criteria is recommended. PMID- 11892016 TI - Discrete epithelioid cells: useful clue to Hodgkin's disease cytodiagnosis. AB - Diagnosis of Hodgkin's disease is made with more confidence than diagnosis of non Hodgkin's lymphoma on cytology. This study was undertaken to describe the presence of granulomas and a new cytologic feature, discrete epithelioid cells (DECs), in smears from Hodgkin's disease. Fine-needle aspiration smears from 39 cases of biopsy-proven Hodgkin's disease, collected over a period of 43 months, were reviewed. Epithelioid granulomas were seen in 38.5% of the smears. DECs, which were noted in 70% of the cases, may be useful in raising the suspicion of Hodgkin's disease when other features are not evident; their presence should encourage the cytopathologist to obtain aspirations from other lymph nodes. PMID- 11892017 TI - Cytology of bone lesions by intraoperative sampling during fracture treatment. AB - Cytology was performed on 314 patients who were treated by surgery for hip joint fracture, to determine and evaluate the role, accuracy, and perspective of intraoperative bone sampling. Specimens were collected from bone lesions during surgery by imprints or driller washing in normal saline. The results were compared with those of subsequent biopsies or clinical follow-up. All 13 neoplastic cases (malignant or benign) were identified by cytology. An accuracy rate of 69.2% was achieved by this method when the type and origin of the neoplasms were to be conclusive. There were no false-positive diagnoses, and all benign conditions showed negative results on cytology (specificity and sensitivity of 100%). Cytology can play a valuable role in the diagnosis of bone lesions. The morphologic diagnostic criteria allow for a high level of diagnostic accuracy of cytologic assessments in most cases of bone lesions, no matter the sampling technique. PMID- 11892018 TI - Fine-needle aspiration cytology of pulmonary rheumatoid nodule: case report and review of the major cytologic features. AB - Patients with rheumatoid disease may develop extra-articular lesions, affecting viscera and soft tissues. Pulmonary rheumatoid nodules show morphologic features reminiscent of a necrotizing inflammation of rheumatoid synovitis and differ from subcutaneous rheumatoid nodules only by their location, extension, and size. Although cytologic studies on pleural effusions in rheumatoid disease have long been performed, there are no more than three reports concerning the fine-needle aspiration (FNA) diagnosis of pulmonary rheumatoid nodules. The authors report on a case of a 62-yr-old woman with rheumatoid disease in whom a FNA diagnosis of pulmonary rheumatoid nodule was successfully performed. The authors highlight the main cytologic features of the entity and emphasise the high index of clinical and pathologic suspicions needed to be able to diagnose this lesion. PMID- 11892019 TI - TIA-1+ cytotoxic large T-cell lymphoma of the mediastinum: case report. AB - A 52-year-old previously healthy Caucasian woman presented with superior vena cava syndrome, secondary to compression of a bulky anterior mediastinal mass involving the right lung. Fine-needle aspiration biopsy of the mediastinum yielded large epithelioid cells intermingled with small mature lymphocytes. The epithelioid cells are LCA positive, expressing cytoplasmic CD3 diffusely and TIA 1 focally, but negative for EMA, CD4, CD8, CD15, CD20, CD30, and CD56. The TIA-1+ cytoplasmic granules correlated to the azurophilic granules in Diff-Quik-stained cells, pink granules in Ultrafast Papanicolaou-stained cells, and dense core granules in electron microscopy. In situ hybridization for Epstein-Barr viral RNA was negative. The background small lymphocytes were composed of a majority of CD4+ T-lymphocytes and minority of CD8+ T-lymphocytes. The patient responded well to six cycles of CHOP chemotherapy, followed by radiation with a total dose of 4140 cGy delivered to the mediastinum in 23 fractions. On the chest X-ray taken 6 mo later, there was minimal apical fibrosis with no evidence of an acute intrathoracic pathology. To the best of our knowledge, this case may be the first report of cytotoxic large T-cell lymphoma of the mediastinum. PMID- 11892020 TI - Microfilariae of Wuchereria bancrofti in cyst fluid of tumors of the brain: a report of three cases. AB - Microfilariae of various nematodes, including Loa loa, Dirofilariae, and Onchocerca volvulus, have been identified in the central nervous system (CNS). The CNS, however, is a rare site for the isolation of microfilariae of Wuchereria bancrofti. To the best of our knowledge, the presence of microfilariae of W. bancrofti in tumor cyst fluids or cerebrospinal fluid has not been reported to date. We report three cases in which microfilariae were identified in the cyst fluid of tumors of the brain. Cyst fluid aspirated from space-occupying lesions in the thalamus and C6-D1 spinal segments in a 46-yr-old man and a 35-yr-old man, respectively, showed numerous microfilariae of W. bancrofti, along with fragments of tumor suggestive of glioma. In the third case, in a 12-yr-old boy, the fluid from the space-occupying lesion in the third ventricle showed microfilariae in a necrotic dirty background with a few squames and cholesteral crystals. Histopathologic examination of the tumor showed an anaplastic astrocytoma and a low-grade astrocytoma in the first two cases, respectively, and a craniopharyngioma in the third case. No microfilariae were identified on the histology sections. PMID- 11892021 TI - Cytopathologic diagnosis of pulmonary sclerosing hemangioma. AB - There are many reports of sclerosing hemangioma from the perspective of its histopathologic features, but its cytopathologic characteristics are less well known. In this report we present the case of a patient in which the cytologic features firmly established a definitive diagnosis; surgical intervention was warranted only after the lesion had grown over the course of 7 yr of close observation. The cytologic diagnosis requires the identification of a dual cell population. Both populations of tumor cell nuclei are immunoreactive for thyroid transcription factor-1, but caution is warranted because this marker may be present in other tumors. Recognition of its distinctive cytologic features can lead to proper diagnosis and conservative management. PMID- 11892022 TI - Dysphagia in a patient with a history of large B-cell lymphoma: esophageal disease with negative biopsy findings. AB - A patient with a previous diagnosis of lymphoma showed signs of dysphagia. Endoscopy found a lesion of the esophagus. Brush cytology and biopsy sampling were accomplished. The biopsy showed inflammation and granulation tissue but no tumor. The cytology specimen, however, was diagnostic of lymphoma. This case emphasizes the need for obtaining cytologic specimens concurrently with biopsies of esophageal lesions. PMID- 11892023 TI - gamma/delta peripheral T-cell lymphoma of the breast diagnosed by fine-needle aspiration biopsy. AB - Gamma/delta T-cell lymphoma is a rare neoplasm that is not well characterized and is associated with a poor prognosis. We report a case of gamma/delta peripheral T cell lymphoma that appeared as a breast lump in a 35-yr-old woman. The patient was examined for a 2-mo history of a right-sided breast mass with associated hepatosplenomegaly 2 yr in duration. A fine-needle aspiration biopsy (FNAB) was performed, and the diagnosis of lymphoma was rendered. The patient received two cycles of CHOP and is alive with persistent disease. FNAB showed evidence of polymorphous lymphoma, consisting of medium-size to large cells with immature chromatin. Flow cytometric immunophenotyping showed expression of CD2, CD3, and CD7 with lack of expression of CD1a, CD4, CD5, CD8, and CD56. Flow cytometry also showed predominant expression of the gamma/delta T-cell receptor. Cytogenetic analysis showed 48XX+i7(q11.2),+7(3). Our case indicates that gamma/delta peripheral T-cell lymphoma can be diagnosed by FNAB. This rare entity requires further investigation. PMID- 11892025 TI - Cytologic features of high-grade squamous intraepithelial lesions involving endocervical glands on ThinPrep cytology. AB - Usage of liquid-based cytology has resulted in better cellular preservation with enhancement of nuclear features. The purpose of this retrospective 2-yr study (January 1999 through December 2000) was to evaluate the cellular features of endocervical gland involvement by a high-grade squamous intraepithelial lesion (HSIL) vs. endocervical adenocarcinoma in situ (AIS) on cell samples processed by the ThinPrep method as compared to conventional smears. Of the 97 cases of CIN III diagnosed on cytology, 52 (54%) showed surface endocervical gland involvement by CIN III and form the basis of this study. There were also six cases of endocervical AIS diagnosed on histology with prior cytology. The architectural features of HSIL involving endocervical glands and AIS were similar to those previously reported on conventional smears. A consistent finding of HSIL involving endocervical glands was the loss of central cell polarity and piling within cell groups, a finding not present in AIS. Central cell polarity was maintained in cellular groupings of AIS. In addition to the cellular feature present on conventional smears, micronucleoli were clearly visualized in cells of HSIL involving endocervical glands and prominent nucleoli were present in AIS. Apoptosis and mitoses were clearly visualized in both entities. Endocervical gland involvement by HSIL has characteristic cell patterns and features on liquid based/thin-layer cytology that permit their distinction from AIS. PMID- 11892024 TI - Clear-cell sarcoma diagnosis by fine-needle aspiration: cytologic, histologic, and ultrastructural features; potential pitfalls; and literature review. AB - A definitive diagnosis of clear-cell sarcoma of soft parts (CCSSP) is possible by fine-needle aspiration (FNA) biopsy alone. The aspirates are markedly cellular, consisting predominantly of discohesive cells but also of cohesive cells. The cytoplasm is eosinophilic and eccentric. The nuclei are round and contain macronucleoli. CCSSP should be considered when FNA of a soft-tissue tumor shows uncharacteristically high cellularity and relatively uniform cells with macronucleoli. Cohesion of some tumor cells does not rule out CCSSP. Melanin pigment and cytoplasmic clearing are infrequent and not necessary for the diagnosis. Sufficient material should always be procured for immunohistochemical studies on the cell block. Seven other cases are found in the literature, all correctly diagnosed by FNA. Although it is rare, CCSSP is a highly malignant tumor that can be diagnosed readily by FNA without resorting to incisional biopsy. PMID- 11892026 TI - Atypical glandular cells of undetermined significance (AGUS): clinical considerations and cytohistologic correlation. AB - The diagnoses of atypical glandular cells of undetermined significance (AGUS) made upon evaluation of cervical/vaginal (Pap) smears is examined to ascertain salient clinical and cytologic features that may lead to better characterization of the true nature of these lesions. Prior history of squamous dysplasia, age of the patient, and the occurrence of abnormal microbiopsy tissue fragments are investigated to determine their value in the proper evaluation of AGUS specimens. Of the 86,234 Pap smears submitted to our laboratory during a period of 2 yr, 187 (0.2%) were diagnosed as AGUS. Available follow-up in 128 (69%) cases revealed 54 (42%) significant tissue proven abnormalities, the majority (55%, 30 patients) of which were diagnosed as squamous intraepithelial lesions (SIL). Squamous dysplasia is significantly more common in women younger than 40 (15/18, 83%) and in patients with prior history of SIL (29/30, 97%). In addition, all nine patients diagnosed with endometrial lesions on subsequent histology were older than 40. Age, however, was not a discriminating factor in women proven to have endocervical glandular lesions. Additionally, certain tissue fragment cytomorphologic features were significantly more often observed on follow-up in specific histologic diagnostic categories. The Pap smears of patients diagnosed with SIL were noted to contain tissue fragments composed of both dysplastic squamous and benign glandular cells in 29 of 30 (97%). The presence of two distinct populations of glandular tissue fragments (typical and atypical) was found in the Pap smears of all nine women with endometrial abnormalities and in the smears of most women subsequently diagnosed with endocervical glandular lesions (87%, 13/15). These observations suggest that a more specific and clinically useful Pap smear interpretation other than AGUS is often possible by consideration of the patient's age and prior history along with the correct identification of the type of atypical cells observed in abnormal tissue fragments. PMID- 11892027 TI - Adenomyoepithelioma of the breast: a cytologic dilemma. Report of a case and review of the literature. AB - Adenomyoepithelioma of the breast is a rare benign tumor made up of epithelial and myoepithelial cells. The cytologic features of this lesion are not well defined. This report describes the cytologic features of a case of adenomyoepthelioma characterized by hypercellularity and the presence of many atypical epithelial cells, leading to the erroneous diagnosis of adenocarcinoma. Review of the cytology literature shows that this condition frequently mimics the cytologic features of a number of benign and malignant breast lesions, thus representing not only an important potential pitfall in the diagnosis of carcinoma but also a differential diagnosis to consider in a variety of breast lesions. PMID- 11892028 TI - Microfilaria in thyroid aspirate. PMID- 11892029 TI - FNA contamination. PMID- 11892030 TI - Effectiveness of fine-needle aspiration cytology of breast: analysis of 2,375 cases from northern Thailand. AB - At the Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, 2,375 cases of breast lesions were sampled by fine-needle aspiration (FNA) from 1994-1999. Cytologic diagnoses were: benign (48%), suspicious for malignancy (5%), malignant (15%), and unsatisfactory (32%). Comparison with histology was possible in 721 cases. The diagnoses obtained by FNA showed a sensitivity of 84.4%, specificity of 99.5%, positive predictive value of 99.8%, negative predictive value of 84.3%, false-negative rate of 16.7%, false-positive rate of 0.5%, and overall diagnostic accuracy of 91.3%. We conclude that, in experienced hands, FNA of breast masses is reliable for diagnosis. Assessment of samples at the time of aspiration can reduce the number of inadequate specimens to near zero. Correlation of FNA results with clinical and radiologic findings can identify false-negatives and false-positives, ensuring optimal patient management. Many centers now recommend needle core biopsy instead of FNA. For regions such as ours, the added cost of this test would make it unavailable to many patients, which could delay a diagnosis of breast cancer. We advocate keeping FNA as a first-line diagnostic procedure, at least in areas under economic restrictions, in order to maximize the availability of health care to women with breast disease. PMID- 11892031 TI - Total mesorectal excision and low rectal anastomosis for the treatment of rectal cancer and prevention of pelvic recurrences. PMID- 11892032 TI - Risk factors for morbidity and mortality after colectomy for colon cancer. PMID- 11892035 TI - Addition of intranasal glucocorticoids to standard antibiotic therapy for sinusitis. PMID- 11892036 TI - Inflammatory patterns of allergic and nonallergic rhinitis. AB - Rhinitis is a chronic condition of the nasal mucosa that affects a large segment of the population. The symptoms of rhinitis occur in a variety of sinonasal conditions, which may be broadly classified as allergic (seasonal or perennial) or nonallergic (infectious or a number of noninfectious etiologies) based on the presence or absence of atopy. The cytokine profile and inflammatory patterns underlying these two conditions vary because of certain differences in their pathophysiology as discussed in this review. PMID- 11892037 TI - Prevalence and differential diagnosis of chronic rhinitis. AB - The prevalence and differential diagnosis of rhinitis changes as we progress from birth to senescence. The heavy burden of allergic rhinitis is often overlooked in infants and disregarded in childhood and adolescence. In women, especially during pregnancy, hormonal changes can significantly affect nasal mucosal hyperreactivity and worsen ongoing syndromes. Various types of inflammatory and noninflammatory nonallergic rhinitides become more prevalent in the fifth decade and beyond. The burgeoning elderly population with irritant, atrophic, and medication-related rhinitis will constitute a greater proportion of our practices as the general population ages. PMID- 11892038 TI - Mechanisms of allergic rhinitis. AB - The pathogenesis of allergic rhinitis can be better appreciated by understanding the numerous protective mechanisms available for mucosal defense. The system of TH2 lymphocytes, IgE production, mast cell degranulation, eosinophil infiltration, and resident cell responses are central to our understanding and treatment of allergic rhinitis. Histamine remains preeminent in causing the cardinal symptoms of the immediate allergic reaction: itching, watery discharge, and nasal swelling. Recruitment and activation mechanisms responsible for the late-phase allergic response are also reviewed. PMID- 11892039 TI - Evidence-based treatment of allergic rhinitis. AB - Allergic rhinitis is an extremely common disease worldwide, affecting 10% to 50% of the population. An increasing prevalence of allergic rhinitis over the past decades and its frequent association with asthma have raised concerns about treating the disease appropriately. New knowledge of the pathophysiologic mechanisms underlying allergic inflammation of the airways has resulted in the development of newer and better therapeutic strategies. This review focuses on evidence-based treatment of allergic rhinitis, highlighting the most recent international consensus and evidence-based guidelines on allergic rhinitis. PMID- 11892040 TI - Viral rhinitis. AB - Viral rhinitis is a common, morbid, and costly malady, often complicated by otitis media, sinusitis, and asthma. Current therapies are relatively ineffective and aimed at reducing symptoms rather than moderating underlying mechanisms. Nasal elevations of proinflammatory cytokines track symptom expression during viral rhinitis, and it is hypothesized that these chemicals orchestrate a common response to infection with many different viruses that cause rhinitis. Also, recent evidence supports a role for neurogenic inflammation in the development of complications. Future studies should dissect the role of proinflammatory cytokines and neuropeptides in the expression of symptoms, signs, pathophysiologies, and complications of viral rhinitis. PMID- 11892041 TI - Grading the severity of allergic rhinitis for treatment strategy and drug study purposes. AB - Grading the severity of rhinitis symptoms is essential for the evaluation of drug efficacy. However, a uniform grading system has not been standardized. A review of 44 journal articles and symposia from Japan, Copenhagen, and the World Health Organization indicates major differences among systems. All except the World Health Organization used a sum of rhinitis scores. The lack of standardization limits comparisons of subjective symptom changes after comparable treatments. Corrective measures include the use of evidence-based definitions of independent nasal symptoms, avoidance of biases in evaluation, and establishment of international consensus for a standardized scale in this era of globalization. PMID- 11892042 TI - Effect of pollutants in rhinitis. AB - Allergic rhinitis is a very common disease worldwide and is influenced by both genetic and environmental factors. Exposure to environmental allergens is the most significant environmental factor in development and exacerbation of allergic rhinitis. However, air pollutants that are not allergens may affect allergic inflammation in the nasal airway. The nasal airway possesses a number of defense mechanisms to deal with environmental irritants. This article examines the effect of ozone and particulate air pollution of TH2-type inflammation in the airway and how nasal defenses protect the upper and lower airway from adverse effects of pollutants. PMID- 11892043 TI - Neurogenic mechanisms in rhinosinusitis. AB - Nasal sensory nerve stimulation leads to sensations of pain and congestion and nociceptive nerve axon response-mediated release of substance P that stimulates glandular secretion as an immediate-acting protective mucosal defense. Recruited parasympathetic reflexes cause submucosal gland secretion via muscarinic M3 receptors. Parasympathetic reflexes, sneezing, and other avoidance behaviors rapidly clear the upper airway of offending agents while protecting the lower airways. Dysfunction contributes to allergic, infectious, and other nonallergic rhinitides and possibly sinusitis. Sympathetic arterial vasoconstriction reduces mucosal blood flow, sinusoidal filling, and mucosal thickness, restoring nasal patency. Loss of sympathetic tone may contribute to some chronic, nonallergic rhinopathies. PMID- 11892044 TI - The molecular biology of nasal polyposis. AB - The molecular biologic events in the development of nasal polyps are now becoming unraveled. It appears that eosinophils are the dominant inflammatory cell present in this tissue. The events leading up to the extravasation of eosinophils into the lamina propria nasal polyps are regulated by the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1 beta. These cytokines upregulate very late antigen-4 on the surface of eosinophils and vascular cell adhesion molecule-1 on the surface of the endothelial blood vessel. Chemokines such as RANTES (regulated upon activation, normal T-cell expressed and secreted) and eotaxin are responsible for the movement of eosinophils into the lamina propria of the nasal polyp. The release of major basic protein has an effect on alteration of the epithelial architecture and on the sodium and chloride flux into and out of the apical epithelial cell of the tissue. Finally, the alteration of the surface epithelium results in a defect in the migration of the cystic fibrosis transmembrane regulator protein to the apical surface. These two events, the release of major basic protein from the eosinophil and the alteration of the architecture of the surface epithelium, lead to an increase in sodium absorption and resultant edema: the hallmark of the pathology of the nasal polyp. PMID- 11892045 TI - Fungal rhinosinusitis: diagnosis and therapy. AB - Fungal rhinosinusitis presents in five clinicopathologic forms, each with distinct diagnostic criteria, treatment, and prognosis. The invasive forms are acute fulminant, chronic, and granulomatous ("indolent") invasive fungal sinusitis. The noninvasive forms are fungal ball ("sinus mycetoma") and allergic fungal sinusitis (AFS). AFS is the most common form of fungal rhinosinusitis. Patients with AFS are atopic to aeroallergens including the involved fungal organism, immunocompetent, have nasal polyps and chronic allergic rhinosinusitis, often produce nasal casts, and may occasionally present with proptosis from orbital extension of disease. Sinus CT shows sinus mucosal hypertrophy and often hyperattenuation of sinus contents. Diagnosis is made from surgical histopathology with or without an associated positive surgical sinus fungal culture. The histopathology shows extramucosal allergic mucin that stains positive for scattered fungal hyphae and eosinophilic-lymphocytic sinus mucosal inflammation. Bipolaris spicifera is the most common fungus cultured. The immunopathology of AFS has been shown to be analogous to allergic bronchopulmonary aspergillosis. Treatment requires surgery and aggressive postoperative medical management with close follow-up. Medical treatment includes allergy medications, allergen immunotherapy, and in many cases the addition of oral corticosteroids. Although medical management clearly improves patient outcomes, more studies are needed because AFS recurrence rates remain high. PMID- 11892046 TI - Sinusitis and asthma: associated airway diseases. AB - Sinusitis and asthma often coexist in patients. In fact, these airways disorders are similar histologically, with tissue eosinophils, increased glandular tissue, and edema. Medical or surgical therapy for sinusitis often greatly improves asthma, suggesting that sinusitis may exacerbate asthma. Possible mechanisms by which asthma could be worsened by sinus disease include neural reflex pathways and interference with the important nasal functions of heating, humidification, and filtration. Health professionals treating asthmatic patients should consider sinusitis as a possible underlying cause, in addition to other triggers (e.g., allergic rhinitis and gastroesophageal reflux disease). PMID- 11892047 TI - Surgical intervention for sinusitis in adults. AB - This article briefly reviews the latest developments in the indications for and performance of paranasal sinus surgery. Although the central role of medical therapy in the treatment of inflammatory chronic rhinosinusitis remains essentially unchanged, the past few years have seen a gradual evolution in the indications for, and the expectations of, sinus surgery. Although many controversies still exist in the optimal management of rhinosinusitis, especially regarding the treatment of chronic frontal rhinosinusitis, the long-term beneficial role of functional endoscopic sinus techniques in combination with medical therapy has become firmly established for patients who do not respond well to medical treatment alone. PMID- 11892048 TI - Surgical intervention for sinusitis in children. AB - Pediatric rhinosinusitis represents a common endpoint of many potential etiologic factors, but fixed anatomic obstruction of sinus outflow is relatively unusual in pediatric patients. Surgical therapy is considered when medical therapy for underlying mucosal inflammation fails. Adenoidectomy is usually the first surgical intervention to be considered for young children, with the goal of improving sinus drainage and eliminating a potential source of bacteria. Endoscopic sinus surgery is considered for the small percentage of patients, most commonly those with underlying pulmonary disease, who fail less aggressive treatment measures. Every decision for surgery involves a risk-benefit analysis. PMID- 11892050 TI - Literature alert. PMID- 11892049 TI - The global burden of asthma and allergic diseases: the challenge for the new century. PMID- 11892051 TI - Tacrolimus ointment: advancing the treatment of atopic dermatitis. PMID- 11892052 TI - Effects of allergic dermatosis on health-related quality of life. AB - Allergic dermatosis is a class of immunologic skin diseases manifested as intense itching, which potentially leads to a cycle of skin pain, damage, and infection. Information collected from clinical samples on health-related quality of life shows that some individuals may suffer from poor-quality sleep, physical and emotional distress, and limitations in social functioning. Although many individual factors may moderate the impact of disease on quality of life, disease severity is consistently linked to amount of limitation. There are only sparse data from population surveys in which participants are not selected based on willingness to join a treatment trial for relief of symptoms. This article presents quality-of-life and disease burden data on 559 persons in a community survey who reported signs and symptoms consistent with allergic dermatosis. Quality of life was assessed using the Dermatology-specific Quality of Life (DSQL) questionnaire. Overall, greater disease severity was associated with higher DSQL scores. Noticeable deficits were reported among those who rated their disease as moderate or severe, especially in terms of physical discomfort, sleep disturbance, and negative self-perceptions and emotions. These complaints correlated significantly with out-of-pocket expenses for lotions and emollients to control skin disease. Despite these complaints, the median number of days from the last primary healthcare visit was 453 days, indicating that many dermatosis sufferers are not accessing expert medical care that could alleviate distress. PMID- 11892053 TI - The true value of the TRUE Test for allergic contact dermatitis. AB - The development of the thin layer rapid use epicutaneous (TRUE) Test is a history of cooperation between scientists from academia and industry covering several disciplines: medicine, pharmaceutical chemistry, pharmacology, and statistics. The TRUE Test is today a patch test system with documented stability and allergen content. Allergens are incorporated in a dried-in-gel vehicle, which is coated onto a polyester backing to form a patch. Applied to the skin, the allergens are released when the gel becomes moisturized by transepidermal water. This may seem to be a simple technique, but its development required laborious research and solutions to stability and dosage problems. The test has been clinically standardized with serial dilution tests on sensitized patients and validated in comparative multicenter tests. The test is a significant step towards higher reliability of patch testing. Fifteen years of experience and critical investigations are discussed in this article, as are possible improvements such as expansion of the test with new allergens. PMID- 11892054 TI - The role of patch testing for chemical and protein allergens in atopic dermatitis. AB - Many patients who present for evaluation of allergic contact dermatitis have an atopic diathesis. Although the immunologic basis of atopic dermatitis differs from that of allergic contact dermatitis--and patients with atopic dermatitis are less easily sensitized under experimental conditions--atopic patients do develop allergic contact dermatitis, and patch testing is a valuable part of their medical care. Delayed (7-day) patch test readings are especially important in atopic patients to distinguish allergy from irritancy and to evaluate for steroid allergy. The utility of atopy patch tests to aeroallergens such as dust mite is increasingly recognized; aeroallergens may be the cause of a type of protein contact dermatitis. PMID- 11892055 TI - Autoimmunity in chronic urticaria and urticarial vasculitis. AB - In contrast to acute urticaria, etiology cannot be identified in most cases of chronic urticaria. Recent evidence suggests that a subset of patients with chronic urticaria may have an autoimmune basis for their condition. The demonstration of antithyroid autoantibodies in some patients with chronic idiopathic urticaria (CIU) provides support for an association. However, the discovery of a positive skin test response to intradermal injection of autologous serum in as many as 60% of patients with CIU led to the identification of autoantibodies to IgE and the alpha-chain of the high-affinity IgE receptor, Fc epsilon RI alpha. Additional studies have demonstrated that some of these autoantibodies are capable of releasing histamine from donor basophils and mast cells. This article reviews the literature that addresses a possible autoimmune etiology in a subset of patients with CIU. Urticarial vasculitis is differentiated from chronic urticaria based on clinical features and biopsy findings of leukocytoclastic vasculitis. Most cases of urticarial vasculitis are secondary to an underlying systemic disease. The presence of autoantibodies has also been demonstrated in a subset of patients with primary urticarial vasculitis. This article briefly reviews some of this data. PMID- 11892057 TI - Chronic urticaria: background, evaluation, and treatment. AB - Urticaria and angioedema will affect 15% of the general population during their lifetime, and this remains one of the most vexing cutaneous conditions to evaluate and treat. Patients frequently go from one physician to another in hopes of finding a healthcare provider who can identify the cause and cure the ailment. Physicians treating hives are equally frustrated as they ponder the utility of obtaining a panel of screening laboratory tests that have previously been shown to have a low yield or obtaining selected allergy tests in a group of patients who are no more prone to allergic disease than the general public. This review presents recent information in a clinical context with the aim of aiding the physician in understanding the pathophysiology of urticaria and formulating an intelligent evaluation and treatment plan. PMID- 11892056 TI - Are barrier creams actually effective? AB - This article reviews the current information surrounding the efficacy of barrier creams as a protective measure against contact dermatitis. The principles of the proposed effects of barrier creams on the skin and the experimental and clinical data regarding their efficacy in the prevention of irritant and allergic contact dermatitis are discussed. PMID- 11892058 TI - The genetics of otitis media. AB - There is significant evidence from epidemiologic, anatomic, physiologic, and immunologic studies that susceptibility to recurrent episodes of acute otitis media (OM) and persistent OM with effusion is largely genetically determined. The genetics of OM are most likely complex, i.e., many genes are probably contributing to the overall phenotype. The knowledge of a hereditary component has important implications because closer surveillance of children at risk for OM could result in earlier detection and treatment. Further, once OM susceptibility genes have been identified it may be possible to develop molecular diagnostic assays that could enable the clinician to identify the child at high risk for OM and to develop more focused treatments in the future. PMID- 11892059 TI - Vaccine prevention of acute otitis media. AB - The incidence of acute otitis media (AOM) in infants and young children has increased dramatically in recent years in the United States. AOM often follows upper respiratory tract infections due to pathogens such as respiratory syncytial virus (RSV), influenza virus, and parainfluenza virus (PIV). These viruses cause eustachian tube dysfunction that is critical to the pathogenesis of AOM. Vaccines against these viruses would likely reduce the incidence of AOM. In three previous studies, influenza virus vaccines reduced the incidence of AOM by 30% to 36%. Vaccines to prevent infections with RSV and PIV type 3 are undergoing clinical testing at this time. Streptococcus pneumoniae, nontypeable Haemophilus influenzae (NTHi), and Moraxella catarrhalis are the three most common AOM pathogens. Heptavalent pneumococcal conjugate vaccine is effective in preventing invasive disease and AOM caused by serotypes contained in the vaccine. Vaccine candidates for NTHi and M. catarrhalis are under development. PMID- 11892060 TI - Antibiotic theory in otitis media. AB - Otitis media is currently the most common diagnosis made by clinicians, which has a major impact on managed care. The emergence of resistant bacterial pathogens has caused controversy over the use of antibiotics when acute otitis media (AOM) is diagnosed. All infants with AOM and all older children with severe AOM should be treated with antibiotics, despite concerns about rising rates of resistant bacterial pathogens. Some older children with nonsevere AOM may be candidates for initial observation, although this is not confirmed by clinical trials. Antimicrobial agents are not required for otitis media with effusion of recent onset but may be considered if this effusion becomes chronic; in these cases, tympanostomy tube placement may be indicated. Antimicrobial prophylaxis for prevention of recurrent AOM should be reserved for selected patients because of the possible emergence of resistant organisms. Tympanostomy tube placement is a more reasonable option today. PMID- 11892061 TI - Overview of ocular allergy treatment. AB - A plethora of drugs is available for the treatment of ocular allergy. Traditional treatment includes antihistamine and antihistamine/vasoconstrictor combination eyedrops. These drugs are useful, safe, and readily available. Mast cell stabilizers are safe, effective, and an important component of antiallergic therapy. Nonsteroidal anti-inflammatory drugs also have antiallergic effects. In recent years, drugs with multiple mechanisms of action have proven to be effective antiallergics. These drugs often have mast cell stabilizing, antihistaminic, and anti-inflammatory properties. Corticosteroids are considered to be more potent than other antiallergic drugs, and modifications in their molecular structures have made certain corticosteroids suitable for the treatment of ocular allergy. PMID- 11892062 TI - Eyelid dermatitis. AB - Cosmetic alteration of a patient's orbital skin is a common reason for professional consultation. This review presents the differential diagnosis and recommended evaluation of inflamed eyelids. To better understand the diseases, each is individually addressed clinically, pathogenetically, and therapeutically. It is critical to recognize the lesions correctly and to have full knowledge of the putative clinical disease process. An algorithm for an appropriate work-up for each disease is offered. With this background, a successful therapeutic response can be anticipated. PMID- 11892064 TI - Allergy and athletes. PMID- 11892063 TI - Ocular allergies. AB - Ocular allergic disease affects not only the conjunctivae but also surrounding structures including the eyelids. Allergic diseases of the eyelid include atopic dermatitis, contact dermatitis, and urticaria/angioedema. They must be differentiated from nonallergic eyelid diseases. Allergic diseases of the conjunctivae comprise a spectrum of disorders from common, non-sight-threatening conditions such as seasonal allergic conjunctivitis, perennial allergic conjunctivitis, and giant papillary conjunctivitis to less common and potentially sight-threatening diseases such as vernal keratoconjunctivitis and atopic keratoconjunctivitis. Each of these conditions is mediated primarily by type I hypersensitivity reactions. The clinical manifestations, differential diagnosis, and treatment of these conditions are reviewed in this article. PMID- 11892066 TI - Gene therapy for immunodeficiency. AB - Since the early 1990s, primary immunodeficiency (ID) disorders have played a major role in the development of human gene therapy. Adenosine deaminase (ADA) deficiency was the first disease to be treated with a gene therapy approach in humans, and was also the first condition for which therapeutic gene transfer into the hematopoietic stem cell has been attempted in the clinical arena. A series of encouraging results obtained in chronic granulomatous disease (CGD) patients have followed these pioneer experiments and preceded the very recent and exciting reports of successful genetic correction procedures performed in patients affected with the X-linked form of severe combined immunodeficiency (XSCID). The technical progress made in the field of gene transfer in recent years is mostly responsible for these clinical advances, and will be critical for future development of gene therapy approaches for other forms of IDs. PMID- 11892067 TI - Hematopoietic stem cell transplantation for severe combined immune deficiency. AB - Hematopoietic stem cell transplantation (HSCT) has been the definitive therapy for severe combined immune deficiency (SCID) since the first successful transplant for SCID in 1968. Improvements in the use of HSCT to treat patients with SCID are continuing. For example, during the last 5 years, the first successful in-utero HSCT, and the first success with gene therapy have occurred in patients with SCID. Debate still continues about the role of pretransplantation therapy for SCID patients, and the biology of post-HSCT immune reconstitution is under investigation. PMID- 11892068 TI - Common variable immunodeficiency. AB - Common variable immunodeficiency (CVID) is a common primary immnodeficiency disease, the hallmark of which is hypogammaglobulinemia. Due to the lack of antibodies, patients usually have recurrent bacterial infections, but there are a number of other puzzling manifestations, including inflammatory conditions, autoimmune disease, and the development of lymphomas. Most patients are diagnosed as adults, and delay in the recognition of the antibody defect is common. A number of defects of T cell function and deficits in the memory B cell pool have been identified, but the underlying cause of this defect remains unknown. PMID- 11892069 TI - Wiskott-Aldrich syndrome. AB - Wiskott-Aldrich syndrome (WAS) is an X-linked immunodeficiency characterized by thrombocytopenia with small platelets, eczema, recurrent infections, autoimmune disorders, IgA nephropathy, and an increased incidence of hematopoietic malignancies. The identification of the responsible gene, WASP (Wiskott-Aldrich Syndrome Protein), revealed clinical heterogeneity of the syndrome, and showed that X-linked thrombocytopenia without, or with only mild immunodeficiency and eczema, is also caused by mutations of WASP. The study of WASP and its mutations demonstrates how a single gene defect can cause multiple and complex clinical symptoms. PMID- 11892071 TI - The hyper IgM syndrome. AB - The hyper IgM syndrome is a rare, inherited immune deficiency disorder resulting from defects in the CD40 ligand/CD40-signaling pathway. X-linked hyper IgM is caused by defects in the CD40 ligand gene, while autosomal recessive hyper IgM is caused by defects in the CD40-activated RNA-editing enzyme, activation-induced cytidine deaminase, which is required for immunoglobulin isotype switching and somatic hypermutation in B cells. The loss of interaction between CD40 and its ligand in X-linked hyper IgM results in an impairment of T cell function, of B cell differentiation, and of monocyte function, while only B cell differentiation appears to be affected in autosomal recessive hyper IgM. With genetic defects in the hyper IgM syndrome identified, it is possible to diagnose patients definitely, to perform genetic screening, and to delineate the clinical manifestations of this syndrome. Further research may lead to novel and definitive therapeutic options for patients with hyper IgM syndrome. PMID- 11892070 TI - DiGeorge syndrome/chromosome 22q11.2 deletion syndrome. AB - DiGeorge syndrome is characterized by conotruncal cardiac defects, hypocalcemia, and a hypoplastic thymus. Many, but not all, patients have a heterozygous deletion of chromosome 22q11.2. In its most severe form, it represents a devastating syndrome with high mortality. Patients with severe immunodeficiency are candidates for a thymic transplant or a fully matched bone marrow transplant. Fortunately, the majority of patients with either DiGeorge syndrome or chromosome 22q11.2 deletion syndrome have a mild to moderate immunodeficiency. These patients may develop recurrent infections or autoimmune disease. PMID- 11892072 TI - Assessment and control of fungal allergens. AB - Fungal sensitivity is a significant cause of allergic disease. Understanding the role fungi play in allergic disease, and how to best control exposure among those with allergy, can have important clinical ramifications. PMID- 11892073 TI - Animal allergens and their control. AB - Animal allergens play a significant role in the pathogenesis of asthma and allergic rhinitis, and are potent causes of acute and chronic symptoms. Although cat and dog allergens are the most important, exposure to a wide variety of other furred animals is not uncommon. Recent reports state that 60% to 70% of households in the western world have at least one pet. Because of this significant exposure, hypersensitivity to animals has become increasingly important. This review focuses on the importance of animal allergens, concentrating on cat and dog allergens, but including others as well. It also discusses the pathogenesis, diagnosis, prevention, and management of animal allergy. PMID- 11892076 TI - Epithelial cells and atopic diseases. PMID- 11892074 TI - Cockroach allergens: environmental distribution and relationship to disease. AB - Cockroach allergy has been recognized as an important cause of asthma. Exposure to high levels of cockroach allergens in the home is a major risk factor for symptoms in sensitized individuals. Previously identified allergens from Blatella germanica and Periplaneta americana include Bla g 2 (inactive aspartic proteinase), Bla g 4 (calycin), Bla g 5 (glutathione-S-transferase), Bla g 6 (troponin), the Group 1 cross-reactive allergens Bla g 1 and Per a 1, Per a 3 (arylphorin), and Per a 7 (tropomyosin). The primary site of cockroach allergen accumulation is the kitchen. However, lower levels of allergen can be found in bedding, on the bedroom floor, and in sofa dust. Strategies for decreasing exposure to cockroach have been investigated. The results suggest that a sustained decrease in cockroach allergen levels is difficult to accomplish, even after successful extermination of cockroach populations. The use of recombinant cockroach allergens may lead to the development of new approaches to asthma treatment in the future. PMID- 11892077 TI - Literature Alert.. PMID- 11892075 TI - Recombinant allergens/allergen standardization. AB - Recombinant allergens are genetically engineered isoforms representing allergen molecules from allergen extracts. Immunologic responses of allergic patients toward allergen extracts define the major allergens. For the average allergic patient, the diagnostic sensitivity and treatment efficacy correlate with the concentration of major allergen. Standardization of allergen products (extracts or genetically engineered allergens) can therefore advantageously be performed using a selected recombinant major allergen as a standard. The standardization will furthermore require reagents for which both monospecific, monoclonal, or preferably, recombinant antibodies can be used. Due to differences in the allergenic activity of individual isoallergens and the naturally occurring mixture of isoallergens found in an allergen extract, and due to additional contribution to the allergenic activity from other molecules in the extract, a biologic potency assessment must always be performed as a supplement. This is also the case for a genetically engineered allergen product. PMID- 11892078 TI - Allergen immunotherapy. AB - Allergen immunotherapy plays an important role in the treatment of allergic diseases and asthma. This article is a brief review of the current approaches, including patient and allergen selection, routes of administration, and use of standardized allergen vaccines. New approaches offering potentially useful strategies based on recent studies of T-cell epitopes, cytokines, and anti-IgE and DNA vaccines also are considered. PMID- 11892079 TI - Allergic rhinitis in children. AB - Epidemiologic and pathophysiologic evidence indicates that allergic rhinitis, whether seasonal or perennial, is one piece of a larger atopic clinical picture that often occurs concomitantly with asthma. Allergic rhinitis usually develops during childhood and has a prevalence of up to 40% in the pediatric population. Careful attention to food allergies and the presence of household allergens during infancy and early childhood may limit potential sensitizations. Many antihistamines and topical corticosteroids now are available for the treatment of allergic rhinitis in children, which is all the more important because optimal management may improve quality of life and curtail the development of serious sequelae. PMID- 11892080 TI - Environmental contributions to allergic disease. AB - The environment is a major contributor to allergic disease, and great effort is being expended to identify the chemical pollutants and allergens that make a significant impact. Exposure to high levels of ozone, sulfur dioxide, nitrogen dioxide, and diesel exhaust particles is known to reduce lung function. Studies continue to delineate the role of these particles as adjuvants and carriers of allergens into the respiratory system. Current studies also show the exacerbation of allergic disease through fungal spore inhalation and continue to document the role of pollen in allergic rhinitis. Pollen also was recently associated with asthma epidemics, especially after thunderstorms. Forecasting models currently are being developed that predict the trajectories of pollen dispersal and may allow increased avoidance of dangerous outdoor conditions. PMID- 11892081 TI - Allergic rhinitis and asthma in children: disease management and outcomes. AB - Antihistamines and inhaled glucocorticoids, which can be targeted toward multiple points in the "allergic cascade" underlying allergic rhinitis and asthma, extend the promise of enhanced outcomes in children with allergic rhinitis, asthma, or both. Antihistamine therapy confers significant relief of subjective ratings of seasonal and perennial allergic symptoms (e.g., rhinorrhea, congestion, sneezing, pruritus), whereas topical steroids alleviate such discomfort while also improving objective anatomic and functional indices of nasal patency (e.g., nasal peak inspiratory flow). Youngsters with asthma also experience substantial clinical benefits from inhaled steroids, which improve objective measures of pulmonary function and reduce rescue beta 2-agonists for symptom management and quality-of-life enhancement. This paper reviews recent clinical findings on the role of antihistamines and topical corticosteroids in pediatric allergy and asthma management, as well as the favorable effects of these medications on both objective and subjective health outcomes. PMID- 11892082 TI - Genetic factors underlying gluten-sensitive enteropathy. AB - Genetic epidemiology clearly has shown that there is a genetic predisposition to gluten-sensitive enteropathy (GSE), or celiac disease. The strong genetic component, as determined by the lambda sib (lambda s), has been calculated to lie in the range of 7.5 to 30, based on a 5% to 10% recurrence risk for siblings. Ninety-five percent of northern European patients with GSE carry a particular HLA DQ alpha beta heterodimer. Studies support the concept that the HLA-DQ gene acts as a dominant gene, and they also found that, in addition to HLA-DQ, a second locus within the major histocompatibility complex (MHC) is involved in the predisposition to GSE in the Dutch population. Genome scans conducted so far suggest that MHC and non-MHC loci collectively contribute to disease susceptibility. Since one, and probably even two, gene(s) from the MHC region itself determine at least 40% to 50% of the genetic predisposition to GSE, it is expected that the other loci each contribute only a little to the total genetic variation. The exact role of these additional genes (i.e., whether they are involved in the initiation or the progression of the disease) remains to be determined. PMID- 11892083 TI - Lyme disease: an update. AB - Lyme disease is a multisystem illness caused by the spirochete Borrelia burgdorferi, and it is the most common vector-borne illness in the United States. Lyme disease is also endemic in Europe and Asia. There have been major advances in the field since the disease was first described, including the sequencing of the B. burgdorferi genome; an increase in understanding of the interactions among the spirochete, the tick, and the mammalian host; new and improved laboratory tests; and a vaccine for prevention of the disease. Still, the diagnosis of Lyme disease remains based on history and clinical findings, supplemented by careful use of laboratory tests, and requires that the physician be familiar with the disease's clinical manifestations and the shortcomings of the available diagnostic tests. PMID- 11892084 TI - The pathogenesis of allergic conjunctivitis. AB - The clinical presentation of the various forms of allergic conjunctivitis varies greatly from mild symptoms to severe disease with vision-threatening complications. Although an IgE-mediated type-1 hypersensitivity reaction has been demonstrated or postulated in many types of allergic eye disease, the pathophysiology underlying the allergic conjunctivitides is not fully understood. The variety of currently available treatment options underscores the complexity of the chemical reactions associated with mast cell degranulation and mediator release causing the onset of allergic signs and symptoms. Many of these treatments are merely palliative and do not eliminate the complex immune response initiating the symptoms, so there is a recurrence of disease as soon as the therapy is discontinued. Models of allergic eye disease have significantly aided the discovery of new anti-allergic and anti-inflammatory compounds that can be used safely in the eye. PMID- 11892085 TI - The clinical spectrum of Bruton's agammaglobulinemia. AB - X-linked, or Bruton's, agammaglobulinemia (XLA) was described in 1952 as the congenital inability to form antibodies. Patients were typically infants or young children with recurrent, severe bacterial infections. Other, milder cases of hypogammaglobulinemia were considered "acquired," and often presented later in life. Since the discovery of the defective gene in XLA in 1993, it has been shown that a significant number of male patients with sporadic or acquired hypogammaglobulinemia actually have XLA. We present here a case of atypical XLA and discuss similar cases in the literature. We conclude that any male with hypogammaglobulinemia, regardless of age of presentation, might have XLA. Males with low B-cell numbers are particularly likely to have XLA and should have Bruton's tyrosine kinase levels assessed. PMID- 11892086 TI - The diagnosis and treatment of Whipple's disease. AB - Whipple's disease is a rare, chronic, and systemic infectious disease caused by the ubiquitously occurring bacterium Tropheryma whippelii. For two reasons, the disease represents a good example for documenting the input of modern molecular based techniques into pathogenetic, diagnostic, and therapeutic concepts in clinical medicine. First, the unidentified and uncultivable causative organism has been characterized by novel molecular-genetic techniques. Second, in contrast to other chronic inflammatory disorders, clinical manifestations of T. whippelii infection seem to be based on reduced T-cell helper type 1 (TH1) activity. These findings have led to an improved pathophysiologic understanding of the disease and to new aspects in treatment strategies that are discussed in this paper. PMID- 11892087 TI - Oral tolerance and gut-oriented immune response to dietary proteins. AB - Dietary proteins induce both a local immune response characterized by IgA production and systemic immune response characterized by the production of TH2/TH3 cytokines. By influencing the class of immune response, the gut enhances the efficacy of immunity against proteins that enter the body through the diet while, at the same time, ensuring its own safety. This article reviews and summarizes the field of "oral tolerance," with emphasis on recent advancements. PMID- 11892088 TI - Dust mites: update on their allergens and control. AB - Mites are ubiquitous organisms, and as a result, humans come into contact with mites and mite products in a variety of situations. Molecules from many mite species can induce IgE-mediated reactions. Best known among the allergy-causing mites are the house dust and storage mites. However, allergists should be aware that, in specific situations, contact with products of many other less-known species of mites also may cause IgE-mediated reactions. PMID- 11892089 TI - Occupational allergens. AB - Occupational agents are important in a significant number of respiratory diseases. More than 250 occupational substances have been reported to cause occupational asthma. Occupational allergens are the subset of agents causing occupational diseases through an IgE-mediated mechanism. These allergens may be classified as being of either high or low molecular weight. The more common occupational allergens and the industries at increased risk of exposing workers to these agents are discussed. PMID- 11892090 TI - Mechanisms of eosinophil recruitment and activation. AB - The role of the eosinophil in the pathophysiology of allergy and asthma has been the focus of intense interest during the last two decades. While the presence of eosinophils in humans with allergy and asthma is well established, the precise role of this cell in humans and in animal models is less clear. However, recent developments in research on many organ systems have provided novel insights into the possible underlying role of the eosinophil in both allergic and nonallergic inflammation. This review examines the pathways associated with eosinophil recruitment and activation and discusses these findings, with reference to clinically defined categories. PMID- 11892092 TI - Basophils in airway disease. AB - The inflammatory response that is often associated with asthma is characterized by the recruitment of eosinophils, basophils, and lymphocytes. Until recently, profiling the basophil and defining its functional characteristics have been difficult. With the advent of some new tools, there is a steadily increasing body of information on the presence and potential activities of the basophil. Although the precise role of these cells in airway diseases, such as asthma, remain unclear, relatively accurate enumeration is now possible. Coupled with new insights into cytokine secretion from these cells, a more accurate picture of the dynamics of this specialized form of inflammation is available for refining our hypotheses regarding its regulation. PMID- 11892091 TI - The mechanisms of aspirin-intolerant asthma and its management. AB - Aspirin is one of the most widely used medications in the world. Adverse effects related to aspirin use were described almost concurrently with its first use. The most common side effects are gastrointestinal and renal, but adverse respiratory effects are not uncommon, and approximately 10% of adult asthmatics are aspirin intolerant. Many of these patients present with the so-called aspirin triad of aspirin sensitivity, chronic rhinosinusitis with associated nasal polyposis, and severe asthma. This paper provides a review of recent investigations into the pathogenesis of this process, which have furthered our understanding of the mechanisms and management of aspirin-induced asthma. PMID- 11892093 TI - The use of inhaled corticosteroids in children with asthma. AB - The inhaled corticosteroids (ICSs) are the most effective long-term controllers for the treatment of childhood asthma. There is now substantial controlled clinical trial data to support the efficacy and safety of ICS therapy in infants and young children (6 months to 4 years of age). These data support the use of nebulizer suspension or metered-dose inhalers with valved holding chambers as effective forms of delivery in this age group. Currently, selection of delivery method depends on the comfort of the parent and the cooperation of the child, as well as on which drug the clinician chooses. The ICSs have a favorable safety profile when administered in currently recommended dosages. A transient 0.5- to 2 cm growth delay occurs in prepubescent children but does not appear to affect attainment of predicted adult height. Long-term trials support the existing recommendations of lowering dosage once control is achieved and stopping therapy when the child's asthma is in remission. PMID- 11892094 TI - Glucocorticoid-resistant asthma. AB - Glucocorticoids are currently the most effective anti-inflammatory therapy for asthma. However, a small subset of asthma sufferers do not respond to clinically relevant doses of glucocorticoids and are termed "glucocorticoid resistant." These patients are characterized by increased bronchial hyperreactivity, lower morning peak expiratory flow rates, and a longer total duration of symptoms. The definition of glucocorticoid resistance is arbitrary, and a dosage and duration of oral glucocorticoid therapy that represent a completely adequate therapeutic trial have yet to be established. For research purposes, glucocorticoid-resistant asthma is defined on the basis of a lack of improvement in airway obstruction (FEV1) following a 2-week course of oral glucocorticoid therapy. Glucocorticoid resistance is associated with in vivo and in vitro alterations in cellular responses to exogenous glucocorticoids. We have implicated abnormal regulation of the activator protein I in the molecular mechanism of glucocorticoid resistance, a phenomenon that may be confined to T cells and monocytes. The identification of an alternatively spliced isoform of the glucocorticoid receptor (GR beta) has sparked interest in the functional role of this isoform and its potential involvement in the pathology of glucocorticoid resistance. Alternative therapies for this condition will have to await a better understanding of the mechanisms of glucocorticoid action. PMID- 11892095 TI - The potential of peptide immunotherapy in allergy and asthma. AB - Allergic conditions contribute significantly to the burden of chronic disease in the industrialized world. Current treatments offer varying degrees of palliation. The sole proven disease-modifying strategy, specific or whole-allergen immunotherapy, is limited because of the associated risk of systemic adverse effects, such as anaphylaxis. Short, linear allergen-derived peptides, corresponding to T cell epitopes, offer the possibility of a safer approach as they are capable of inducing allergen-specific hyporesponsiveness without cross linking mast cell-bound IgE. This review evaluates the scientific basis of peptide immunotherapy and clinical experience in allergy up to the present time. PMID- 11892096 TI - The role of nerves in asthma. AB - Asthma is a syndrome characterized by reversible episodes of wheezing, cough, and sensations of chest tightness and breathlessness. These symptoms are secondary to changes in the activity of the nervous system. The mechanisms by which the nervous system is altered such that the symptoms of asthma occur have not yet been elucidated. Airway inflammation associated with asthma may affect neuronal activity at several points along the neural reflex pathway, including the function of the primary afferent (sensory) nerves, integration within the central nervous system, synaptic transmission within autonomic ganglia, and transmission at the level of the postganglionic neuroeffector junction. We provide a brief overview of these interactions and the relevance they may have to asthma. PMID- 11892099 TI - Dendritic cells--the link between innate and adaptive immunity in allergy. PMID- 11892097 TI - Peripheral airways in asthma. AB - The peripheral, or small, airways are usually defined as conducting airways that are less than 2 mm in internal diameter and extend from the noncartilaginous bronchioles to the alveolar ducts. Noninvasively measuring the function of the small airways in isolation is difficult since they make up only about 10% of total airway resistance. Quantitative pathologic studies have shown that both the small and large airways are involved in inflammation and remodeling in asthma. Recent studies also have shown that inflammation involves the alveoli surrounding small airways in asthma and that the distribution of different inflammatory cells across the airway wall varies in both large and small airways. Inhaled treatment that targets the small airways may be more effective than treatment that is deposited more proximally and suggests that treatments in the future need to address the variable distribution of pathology in the bronchial tree in asthma. PMID- 11892100 TI - Literature alert. PMID- 11892098 TI - The role of allergens in the induction of asthma. AB - Despite the strong and consistent association between immediate hypersensitivity and asthma, there is still controversy about the role inhaled allergens play and about the timing of events related to sensitization. Recent studies have provided further evidence on the nature of the immune response to allergens, the timing of this response, and, in particular, whether any response to allergens occurs in utero. Some of the studies also provide a better explanation for why there is not a simple dose response relationship between allergens and asthma. The new studies also raise major issues about the nature of the immune response in nonallergic individuals. Taken together, the findings do not support a simple view about the balance between TH1 and TH2 responses, but strongly support the relevance of IgE to the risk of asthma. PMID- 11892101 TI - Does NATN have any advice or research available about home laundering of scrub attire? PMID- 11892102 TI - Latex facts in perioperative practice. PMID- 11892103 TI - The underpinnings. AB - In last month's Journal, readers were introduced to the government's Improving Working Lives Strategy. This month some of the underpinning detail will be examined in order to show what is expected from employers and employees when getting the balance right between work and life away from work. PMID- 11892104 TI - Effective management. What is it? AB - The first article in this series looked at the importance of reconciling the professional and commercial aspects of our organisations. We found that it is important to develop a better understanding of both by working together to share information and expertise. In this article we will consider what management means in practice and how we can help or hinder its effectiveness. PMID- 11892105 TI - Leadership makes a difference. PMID- 11892106 TI - Don't blame the iceberg. AB - At the above-mentioned study day, Lesley Fudge gave a colleague's paper on risk management. The colleague was Jill Ashley-Jones, then a Health Safety Manager at the Frenchay Hospital at the North Bristol NHS Trust, and she had been unable to attend the day. She had focused on the risk management issues associated with the sinking of the Titanic and had based it on the actual reports written following the disater. There was no mention of Leonardo or Kate, of course. It was also a clever approach when one looks at what happened, and inspired this article by Jim Miller. The mistakes were fundamental and the risk management issues are absolutely classic. Jill associated some of the lessons learnt with what is happening in the NHS, which can, in turn, be applied to a specific team, to a department or to an organisation. PMID- 11892107 TI - How do you keep your patient safe? Biological issues that impact on procedures. AB - In this fourth article in the Concepts in Anatomy series, Mriga Williams discusses the issue of keeping patients safe within the operating theatre department. This series is based on Physiology and Anatomy, a homeostatic approach, John Clancy and Andrew McVicar (eds), Edward Arnold, London, 1995. PMID- 11892108 TI - Mixing systems and the effects of vacuum mixing on bone cement. AB - The aim of a good cement mix is to produce bone cement that has the best mechanical properties possible in order that it can carry out its load transfer role successfully over the lifetime of the implant. The data presented shows that vacuum mixing reduces the cement porosity, which results in improved strength, and resistance to creep deformation and fatigue failure in the bone cement. It seems, however, that eliminating a high degree or all of the cement porosity may be detrimental because it leads to greater shrinkage and cracking in the material. A moderate vacuum level will improve the mechanical properties, but reduce the risk of thermal shrinkage and cracking which is seen at higher vacuum levels. In addition, the mixer design has a significant influence on the quality of the cement produced, affecting porosity, unmixed powder and subsequently the mechanical properties of the material. In the next issue we will be concluding this series of articles by covering the importance of temperature and its effects on the phases of the cement polymerisation process. Bone cement training courses are being run within hospitals by Summit Medical as part of our commitment to enhancing the skills of the perioperative practitioner and to ensuring the best long-term outcome for the patient. PMID- 11892109 TI - Picking peers. PMID- 11892110 TI - Principles of bone cement mixing. AB - This is the first in a series of four consecutive articles on bone cement mixing. It aims to bring to the perioperative practitioners' attention, all of the factors that will influence the quality and reproducibility of the bone cement they produce. The series will demonstrate how perioperative practitioners can have a direct influence on the successful long-term result of a hip or knee replacement. PMID- 11892111 TI - The surgical neonate. AB - This is the third article in the Concepts in Anatomy series, based on Physiology and Anatomy, a homeostatic approach, by John Clancy and Andrew McVicar. In this article, Stevie Boyd discusses some of the problems encountered in neonatal surgery. PMID- 11892112 TI - Advancing perioperative care. AB - Demands are continuing to grow for perioperative practitioners to gain more skills and higher levels of education. These pressures are resulting, in part, from the changing structures within the health service, developments in the medical sciences, the reallocation of roles and responsibilities among healthcare professionals and the recognition by perioperative nurses of their own need for further education to meet these challenges. A response has been the creation of a widening range of new courses giving practitioners opportunities to study at higher degree level. In this article, Paul Wicker and Rebecca Strachan explain how one such programme has been brought into being. PMID- 11892113 TI - The benefits of using customized procedure packs. AB - Discrete item purchasing is the traditional approach for hospitals to obtain consumable supplies for theatre procedures. Although most items are relatively low cost, the management and co-ordination of the supply chain, raising orders, controlling stock, picking and delivering to each operating theatre can be complex and costly. Customized procedure packs provide a solution. PMID- 11892114 TI - Diathermy safety. PMID- 11892115 TI - Latex facts in perioperative practice. PMID- 11892116 TI - We are trying to find the most appropriate way of handling scalpels and other sharps to the surgeon during surgery. PMID- 11892117 TI - Histopathology of cochlear implants. PMID- 11892118 TI - Surgical methods of the Clarion 16 cochlear implant device. PMID- 11892119 TI - Cochlear microendoscopy and CO2 laser vaporization during cochlear implantation. PMID- 11892120 TI - Compressed electrode arrays in cases of obliterative labyrinthitis ossificans. PMID- 11892121 TI - Cochlear implant in inner ear malformation: double posterior labyrinthotomy approach to common cavity. PMID- 11892122 TI - Pitch perception through the cochlear implant for speech and music. PMID- 11892123 TI - Management and control of gusher during cochlear implant surgery. PMID- 11892124 TI - Cochlear reimplantation. PMID- 11892125 TI - 'Veria operation': cochlear implantation without a mastoidectomy and a posterior tympanotomy. A new surgical technique. PMID- 11892126 TI - Facial nerve anomalies in cochlear implantation. PMID- 11892127 TI - Cochlear implant in a child with totally ossified cochlea. PMID- 11892128 TI - Cochlear implant for a totally deaf patient with superficial hemosiderosis of the central nervous system. PMID- 11892129 TI - A mother and son cochlear implant case study: making the decision twice. PMID- 11892130 TI - Factors influencing speech perception abilities in cochlear-implanted children. PMID- 11892131 TI - The role of round window electrophysiological techniques in the selection of children for cochlear implants. PMID- 11892132 TI - Auditory responses of field L to electrical stimulation. PMID- 11892133 TI - A novel outer ear canal electrode for noninvasive electrical stimulation of the cochlear nerve in preoperative evaluation for cochlear implants in adults and children. PMID- 11892134 TI - Cochlear implantation in a patient with Cogan's syndrome, chronic ear disease and on steroid therapy. PMID- 11892135 TI - Cochlear implantation in auditory neuropathy. PMID- 11892136 TI - Reliable test conditions for the vowel-consonant confusion test in cochlear implantees. PMID- 11892137 TI - Use of stapedius reflex in cochlear implant of congenitally deaf children. PMID- 11892138 TI - Vestibular assessment and ear selection for cochlear implantation: the role of bedside testing. PMID- 11892139 TI - Dizziness and vertigo after cochlear implantation. PMID- 11892140 TI - Significance of vestibular function in cochlear implant patients. PMID- 11892141 TI - Intracellular recording from the primary auditory cortex. PMID- 11892142 TI - A case of CSF gusher in Mondini deformity of the inner ear. PMID- 11892143 TI - Vestibular function in patients with cochlear implant surgery. PMID- 11892144 TI - Clinical experience with partially implantable middle ear implant. PMID- 11892145 TI - The phenomenon of nystagmus upon electrical stimulation in a cochlear implant patient. PMID- 11892146 TI - Factors influencing speech development in infants with cochlear implants. PMID- 11892147 TI - Surgical concepts for cochlea implantation in young and very young children. PMID- 11892148 TI - Results from 88 prelingually deaf children with cochlear implants: an analysis of predictive factors. PMID- 11892149 TI - Effects of the programed teaching materials of speech and hearing rehabilitation on children with cochlear implants and hearing aids in Taiwan. PMID- 11892150 TI - Longitudinal communication skill acquisition in pediatric cochlear implant recipients. PMID- 11892151 TI - Cochlear implant in a young child with Waardenburg syndrome. PMID- 11892152 TI - Bilateral cochlear implant--new aspects for the future? PMID- 11892153 TI - Cochlear implantation in Chinese children. PMID- 11892154 TI - Functional magnetic resonance tomography of the auditory cortex during noninvasive stimulation of the cochlear nerve. PMID- 11892155 TI - Electrical stimulation of the auditory pathway in deaf patients following acoustic neurinoma surgery and initial results with a new auditory brainstem implant system. PMID- 11892156 TI - Auditory brainstem implant (Nucleus 21-channel) in neurofibromatosis type 2 patients previously operated on: preliminary results. PMID- 11892157 TI - Experience with cochlear implants in postmeningitic cases. PMID- 11892158 TI - The Canossian School for the Hearing Impaired-Singapore General Hospital (CSHI SGH) Cochlear Implant Program: children's auditory speech perception scores pre- and postimplant. PMID- 11892159 TI - The speech and language rehabilitation program for pediatric cochlear implantees in Hong Kong. PMID- 11892160 TI - Outcome after cochlear implantation and auditory verbal training in terms of speech perception, speech production and language. PMID- 11892161 TI - What makes cochlear implant patients anxious? PMID- 11892162 TI - Effects of the number of active electrodes on tone and speech perception by Nucleus 22 cochlear implant users with SPEAK strategy. PMID- 11892163 TI - Using electrically evoked compound action potential thresholds to facilitate creating MAPs for children with the Nucleus CI24M. PMID- 11892164 TI - Change of the consonant phonation ability in cochlear implantees. PMID- 11892165 TI - Relationship of the implant electrical auditory brainstem response threshold to the postoperative T level in children with cochlear implants. PMID- 11892166 TI - Satisfaction with the cochlear implant of pre- and postlingually deaf adults. PMID- 11892167 TI - Influences of variables for communicative targets after cochlear implantation. PMID- 11892168 TI - Clinical application of magnetic resonance imaging in cochlear-implanted patients. PMID- 11892169 TI - Hearing aid optimization in the evaluation of cochlear implant candidacy. PMID- 11892170 TI - Development of play in a young cochlear implant user. PMID- 11892171 TI - A follow-up study of a congenitally deaf child with a cochlear implant by a multidisciplinary team. PMID- 11892172 TI - Loudness scaling from preoperative testing to fitting cochlear implants and auditory brainstem implants. PMID- 11892173 TI - Individual auditory skill profile as an original method of demonstrating auditory skill development. PMID- 11892174 TI - Language and reading acquisition in children with and without cochlear implants. AB - These data provide a coherent view of the spoken language and reading skills of children receiving cochlear implants. The data reveal both spoken language gains and clear benefit to these children with regard to reading. Improvement in reading is consistent with predictions based on prior research demonstrating a strong association between spoken language and reading. An intervention such as a cochlear implant has a direct effect on spoken language, and this can subsequently affect reading performance. This provides some of the first experimental evidence supporting the causal relationship between spoken language and reading. PMID- 11892175 TI - 7-year speech perception results and the effects of age, residual hearing and preimplant speech perception in prelingually deaf children using the Nucleus and Clarion cochlear implants. PMID- 11892176 TI - Evaluation of different choices of n in an n of m processor for cochlear implants. PMID- 11892177 TI - Intraoperative stapedius reflex measurements after deep electrode insertion compared with postoperative fitting values in cochlear implant patients. PMID- 11892178 TI - Cortical activity and speech perception performance in cochlear implant users. PMID- 11892179 TI - Hearing performance of cochlear implant patients in noise. PMID- 11892180 TI - Speech discrimination scores of postlingually deaf adults implanted with the Combi 40 cochlear implant. PMID- 11892181 TI - Cochlear implantation in a child with bilateral large vestibular aqueducts. PMID- 11892182 TI - Comparison of hearing ability in cochlear implant subjects with normal and ossified cochlea. PMID- 11892183 TI - Cochlear implants: a 30-year case study. PMID- 11892184 TI - Changes in electrical thresholds and dynamic range over time in children with cochlear implants. PMID- 11892185 TI - Sound and speech recognition ability in Korean cochlear implantees, prelingually deaf children. PMID- 11892186 TI - New electrode concepts (modiolus-hugging electrodes). PMID- 11892187 TI - The Nucleus Double Array cochlear implant for obliterated cochleae. PMID- 11892188 TI - Topographic analysis of inner ear lesions in patients with cochlear implantation using three-dimensional magnetic resonance imaging. PMID- 11892189 TI - Bilateral cochlear implantation--a case report. PMID- 11892190 TI - The Manchester Adult Cochlear Implant Programme 1988-1999. PMID- 11892191 TI - Speech discrimination in elderly cochlear implant users. PMID- 11892192 TI - Basic study for the sentence perception ability of hearing-impaired and cochlear implanted children. PMID- 11892193 TI - Speech perception and production performance of prelingually deafened adolescents after cochlear implantation. PMID- 11892194 TI - Economic evaluation of cochlear implantation. PMID- 11892195 TI - Results of cochlear implant in postlinguistically deaf adults. PMID- 11892197 TI - Residual hearing following a cochlear implantation: effect of time and device. PMID- 11892196 TI - Residual hearing after cochlear implantation. PMID- 11892198 TI - Magnetic resonance imaging safety of the Combi 40/Combi 40+ cochlear implants. PMID- 11892199 TI - Cochlear implantation in the elderly. PMID- 11892200 TI - Changes in residual hearing after cochlear implantation. PMID- 11892201 TI - Fast stimulator cochlear implants in patients with residual hearing. PMID- 11892203 TI - Benefit of ACE compared to CIS and SPEAK coding strategies. PMID- 11892202 TI - Influence of the band-pass filter configuration on speech perception and patient preference in the Combi-40+ cochlear implants. PMID- 11892204 TI - Study on the effect of the number of electrodes programmed and the area of stimulation in cochlear implant patients. PMID- 11892205 TI - Speech perception results in children implanted with the CLARION Multi-Strategy cochlear implant. PMID- 11892206 TI - Brain function of cochlear implant users. PMID- 11892207 TI - Speech perception results in adults implanted with the CLARION Multi-Strategy cochlear implant. PMID- 11892208 TI - Comparison of postlingually deafened adults using the SPEAK and CIS coding strategies. PMID- 11892209 TI - Comparison of the results of two different implant systems using Continuous Interleaved Sampler speech processing strategy. PMID- 11892210 TI - Cochlear implantation in Iran: a report of 190 cases. PMID- 11892211 TI - Evaluation of the performance in the Clarion cochlear implant users in Japan. PMID- 11892212 TI - Initial results of patients implanted in the MED-EL Combi 40+ investigational trial at Virginia Mason Medical Center. PMID- 11892213 TI - 3-dimensional fast spin-echo in determining the indication for cochlear implantation. PMID- 11892214 TI - Cerebral cortical hypometabolic area in 18F-FDG positron emission tomography inversely relates to the duration of deafness in prelingually deaf patients. PMID- 11892215 TI - Development and plasticity of the auditory cortex in cochlear implant users: a follow-up study by positron emission tomography. PMID- 11892216 TI - Sign language activated the auditory cortex of a congenitally deaf subject: revealed by positron emission tomography. PMID- 11892217 TI - Regional cerebral activation during auditory stimulation in a patient with binaural cochlear implants using 99mTc-ECD SPECT. PMID- 11892218 TI - Image study on cochlear-implanted patients. PMID- 11892219 TI - Expression of glutamate receptors in the cochlea of the normal and kanamycin-deaf rats. PMID- 11892220 TI - Valsalva-induced scalp flap elevation after cochlear implantation: etiology and treatment. PMID- 11892221 TI - A surgical technique for cochlear implantation in very young children. PMID- 11892222 TI - Fixation of intracochlear electrode at the cochleostomy. PMID- 11892223 TI - The modified split bridge method to prevent electrode slip-out. PMID- 11892224 TI - Cochlear implants in children (soft surgery). PMID- 11892225 TI - Cochlear implant in children with common cavity deformity. PMID- 11892226 TI - Cochlear implant surgery in ears with chronic otitis media. PMID- 11892227 TI - Cochlear implantation surgery: problems and solutions. PMID- 11892228 TI - Malpractice: why you may be forced to settle. PMID- 11892229 TI - How I cracked the CPT codes. PMID- 11892230 TI - Calming the waters in group practice conflicts. PMID- 11892231 TI - Sarah's last visit. PMID- 11892232 TI - Could you climb out of this financial hole? PMID- 11892233 TI - What makes a group practice thrive. PMID- 11892234 TI - Does your compensation formula pass the smell test? PMID- 11892235 TI - My best quit-smoking aid: Philip Morris. PMID- 11892236 TI - 8 ways to escape your employer. PMID- 11892237 TI - Federation analysis reveals $158 billion "stealth cut" to Medicare. PMID- 11892238 TI - MedPAC recommends full marketbasket for rural and small urban hospitals. PMID- 11892239 TI - If nothing else, give children a healthy start in life and improve Medicare rural health. PMID- 11892240 TI - Fibre types in skeletal muscles. AB - This book is a concise summary of the present knowledge about skeletal muscle fibres. The fibre types were characterized from different points of view. The difficulties and possibilities of classifying muscle fibres in distinct non overlapping types were shown. A main emphasis is put on metabolic fibre typing by cytophotometry as well as the adaptability of a given fibre type to altered physiological and pathological conditions. Extensive analyses of several rat hindlimb muscles revealed regional differences of fibre properties within the muscles and showed the influence of ageing, myopathy, hypoxia, diabetes and Ginkgo biloba-treatment on the different fibre types. The plasticity of muscle fibres was demonstrated by the flexibility of fibre metabolism as an adaptive mechanism. PMID- 11892241 TI - Fundamental structural aspects and features in the bioengineering of the gas exchangers: comparative perspectives. AB - Over its life, an organism's survival and success are determined by the inventory of vital adaptations that its progenitors have creatively appropriated, devised and harnessed along the evolutionary pathway. Such conserved attributes provide the armamentarium necessary for withstanding the adverse effects of natural selection. Refinements of the designs of the respiratory organs have been critical for survival and phylogenetic advancement of animal life. Gas exchangers have changed in direct response to the respiratory needs of whole organisms in different environmental states and conditions. Nowhere else is the dictum that in biology 'there are no rules but only necessities' more manifest than in the evolutionary biology of the gas exchangers. The constructions have been continually fashioned and refined to meet specific needs. Solutions to common respiratory needs have been typified by profound structural convergence. Over the evolutionary continuum, as shifts in environmental situations occurred, infinitely many designs should theoretically have emerged. Moreover, without specific selective pressures and preference for certain designs, considering that there are only two naturally occurring respirable fluid media (air and water), air-lungs, water-lungs, air-gills and water-gills would have formed to similar extents. Factors such as body size, phylogenetic level of development, respiratory medium utilized and habitats occupied have permutatively prescribed the design of the gas exchangers. The construction of the modern gas exchangers has eventuated through painstaking cost-benefit analysis. Trade-offs and compromises have decreed only a limited number of structurally feasible and functionally competent outcomes. The morphological congruity (analogy) of the gas exchangers indicates that similar selective pressures have compelled the designs. Solutions to metabolic demands for molecular O2 have only differed in details. Passive physical diffusion, for example, is the ubiquitous method of transfer of O2 across biological tissues. Gills, evaginated gas exchangers, were the primordial respiratory organs that evolved for water breathing, whereas lungs (invaginated gas exchangers) developed for terrestrial (air) breathing. Transitional (= bimodal = amphibious) breathing has evolved in animals with specialized organs that extract O2 from both water and air. Lungs are tidally (= bidirectionally) ventilated, while gills are unidirectionally ventilated, a feature that allows the highly efficient counter-current disposition between blood and water. Since animals occupy inconstant environmental milieus and their metabolic states vary, gas exchangers are designed to operate optimally across a spectrum of conditions that range from resting to exercise and even under hypoxia. Inbuilt structural and functional flexibility provides the requisite safety factors that allow adjustments to modest pressures. The fundamental structural features that determine the respiratory function of a gas exchanger are respiratory surface area, thickness of the blood-water/gas (tissue) barrier and volume of the pulmonary capillary blood. The diffusing capacity of a gas exchanger correlates directly with the surface area and inversely with the thickness of the blood-water/gas (tissue) barrier. An extensive surface area is generated in gills by extensive stratification of the gas exchanger and in lungs by profuse internal subdivision. Compartmentalization yields small terminal gas exchange compartments that compel greater commitment of energy to ventilate. The surfactant, a phospholipid lining, reduces the forces of surface tension at the air-water interface. This attenuates the propensity of physical collapse of the minute gas exchange units and minimizes the cost of ventilation. The surfactant characterizes all the gas exchangers derived from the piscine air bladder. In the lower air-breathing vertebrates, such as the lungfishes (Dipnoi), amphibians and certain reptiles, the pneumocytes are not differentiated into type I and II cells, as is the case in the lungs of the higher vertebrates-birds and mammals. It is envisaged that in endotherms, the overall numerical density of the pneumocytes and hence the O2 consumption of the gas exchangers may be reduced and a thin blood-gas (tissue) barrier generated, factors that enhance respiratory efficiency. The thin blood-gas (tissue) barriers, for example, those of the mammalian (in the respiratory sections of the interalveolar septum) and avian lungs, consist of an epithelial cell and an endothelial cell with a common basement membrane. An interstitial space occurs in the blood-air/water (tissue) barriers of the gas exchangers of fish gills and lungs of lungfishes, amphibians, reptiles and in the supportive parts of the interalveolar septum of the mammalian lung. Collagen, elastic tissue, nerves, lymphatic vessels and smooth muscle elements are found in the interstitial space. The thickness of the blood air/water (tissue) barrier allometrically changes very little. This suggests that the thicknesses of the blood-water/air (tissue) barriers have been optimized. The presentation and exposure to the gas exchange media (water/air to blood), features dictated by the geometry and arrangements of the structural components of the gas exchangers, contribute greatly to respiratory efficiency. The countercurrent presentation between water and blood in fish gills is the most efficient design in the evolved gas exchangers: It was imperative for survival in water, a medium that contains relatively less O2 and is more expensive to breathe. In the evolved vertebrate gas exchangers, the exposure of blood to air is best manifested in the diffuse design of the avian lung, where the capillary blood is literally suspended in a three-dimensional air space, the blood being exposed to air virtually across the entire blood-gas (tissue) barrier. A double capillary design occurs in the lungs of amphibians and generally those of reptiles, whereas a single capillary design commonly occurs in those of adult mammals. The capillary loading (the ratio of the volume of the capillary blood to the surface area across which blood is exposed to air) in lungs with a double capillary arrangement is high and manifests a poor design. On the other hand, the low capillary loading that characterizes the single capillary system indicates better exposure of blood to air and greater respiratory capacity. Fractal geometry features in the construction of the gas exchangers. The highly versatile design allows the gas exchangers to function optimally under different conditions and circumstances and to maintain congruent morphologies over a wide range of body size, shape and metabolic capacities. At the gas exchange level, sheet-flow design preponderates in the evolved gas exchangers; blood is efficiently exposed to the external respiratory medium. The respiratory capacity of a gas exchanger is comprehensively granted by refinements of structural features and functional processes. Modelling, mathematical integration of structural and functional parameters, provides a holistic view of the essence of the design of a gas exchanger. PMID- 11892242 TI - Molecular biology of lentivirus-mediated gene transfer. PMID- 11892243 TI - Production of lentiviral vectors. PMID- 11892244 TI - Biosafety issues in lentivector production. PMID- 11892245 TI - Lentiviral vector targeting. PMID- 11892246 TI - Integration site selection by lentiviruses: biology and possible control. AB - Retroviruses integrate into naked DNA in a generally sequence nonspecific fashion, but closer study reveals a variety of forces that influence target site selection. Primary sequence of the target plays a small but detectable role. Proteins bound to target DNA can inhibit integration by blocking access of integration complexes or stimulate integration by distorting DNA. An important example of the latter is DNA distortion in nucleosomal DNA. In vivo integration has not yet been convincingly shown to be biased in favor of any identifiable sequence features, though this could still change in future studies. Many applications of retroviral vectors could be facilitated by targeting integration in vivo to predetermined sites. Towards this end, several groups have studied the properties of fusions of integrase proteins to sequence-specific DNA-binding domains. To date such studies establish that targeting can work well in reactions in vitro, but a variety of obstacles complicate applications in vivo. However, naturally occurring retrotransposons do carry out highly targeted integration using retrovirus-like integrase proteins, fueling long-term hopes for targeting with retroviral integrases as well. PMID- 11892247 TI - Improving the post-transcriptional aspects of lentiviral vectors. PMID- 11892248 TI - Lentiviruses as vectors for CNS diseases. PMID- 11892249 TI - Lentiviral vectors for the gene therapy of lympho-hematological disorders. PMID- 11892250 TI - Lentiviral vectors for gene therapy of HIV-induced disease. PMID- 11892251 TI - Opportunities for the use of lentiviral vectors in human gene therapy. PMID- 11892252 TI - HIV-1-derived lentiviral vectors. PMID- 11892254 TI - Nonprimate lentiviral vectors. PMID- 11892253 TI - Lentiviral vectors derived from simian immunodeficiency virus. PMID- 11892255 TI - Biaryls in nature: a multi-facetted class of stereochemically, biosynthetically, and pharmacologically intriguing secondary metabolites. PMID- 11892256 TI - The naturally occurring coumarins. PMID- 11892257 TI - Rheumatic disorders and glucocorticoid-induced osteoporosis. PMID- 11892258 TI - Epidemiology of glucocorticoid-induced osteoporosis. PMID- 11892259 TI - Treatment of glucocorticoid-induced osteoporosis with calcium, vitamin D and D metabolites. PMID- 11892260 TI - DXA in the diagnosis of glucocorticoid-induced osteoporosis. PMID- 11892261 TI - Therapeutic potentials of androgens, estrogen and SERMs in glucocorticoid-induced osteoporosis. PMID- 11892262 TI - Calcitonin in glucocorticoid-induced osteoporosis. PMID- 11892263 TI - Bisphosphonates in the treatment of glucocorticoid-induced osteoporosis. PMID- 11892264 TI - Fluoride and anabolic steroids in the treatment of glucocorticoid-induced osteoporosis. PMID- 11892265 TI - Growth hormone in glucocorticoid-induced osteoporosis. PMID- 11892266 TI - PTH in the pathogenesis and treatment of glucocorticoid-induced osteoporosis. PMID- 11892267 TI - The cellular and molecular basis of glucocorticoid actions in bone. PMID- 11892268 TI - The use of quantitative ultrasonometry in patients with glucocorticoid-induced osteoporosis. AB - There is a great need to introduce methods to determine bone fragility in outpatient settings for chronically GC-treated patients which are easy to use, reasonably inexpensive, precise and have a high sensitivity and specificity to detect patients at risk for fractures. DXA and QUS have both been shown to predict hip fracture rates independently in prospective studies in postmenopausal women. No such studies have been done for patients under GC treatment yet. Most studies investigated patients with established GCO and only one large study has investigated patients with risk factors such as intermittent GC intake and compared DXA and QUS. No studies have been done using QCT. All studies show that QUS is equally potent in detecting patients at risk compared to DXA. There is good evidence that phalangeal QUS is highly dependent on structural changes of trabecular bone. Phalangeal QUS might thus have the advantage of detecting GC induced structural deterioration earlier then calcaneal devices, however no clinical comparison studies have been conducted. The best would be to do longitudinal studies to see which device and site best predicts bone loss, however this requires large patient numbers and patients at risk are usually in different departments due to the difference in underlying diseases. PMID- 11892269 TI - Bone histomorphometry in untreated and treated glucocorticoid-induced osteoporosis. PMID- 11892270 TI - Biochemical markers in glucocorticoid-induced osteoporosis. PMID- 11892271 TI - Osteoporosis in Cushing's syndrome. PMID- 11892272 TI - Adrenal incidentalomas and subclinical Cushing's syndrome: is there evidence for glucocorticoid-induced osteoporosis? PMID- 11892273 TI - Glucocorticoid-induced osteoporosis in asthma and respiratory diseases. PMID- 11892274 TI - Organ transplantation and glucocorticoid-induced osteoporosis. PMID- 11892275 TI - An overview of actin-based calcium regulation. PMID- 11892276 TI - Cooperativity in the Ca2+ regulation of muscle contraction. PMID- 11892277 TI - Motility assays of calcium regulation of actin filaments. PMID- 11892279 TI - The role of troponin in the Ca(2+)-regulation of skeletal muscle contraction. PMID- 11892278 TI - The ultrastructural basis of actin filament regulation. PMID- 11892280 TI - Structural changes between regulatory proteins and actin: a regulation model by tropomyosin-troponin based on FRET measurements. PMID- 11892281 TI - Fluorescence resonance energy transfer in acto-myosin complexes. PMID- 11892282 TI - Insights into actomyosin interactions from actin mutations. PMID- 11892283 TI - Role of charges in actomyosin interactions. PMID- 11892284 TI - The alanine-scanning mutagenesis of Dictyostelium myosin II at the ionic interface with actin. PMID- 11892285 TI - Changes in actin and myosin structural dynamics due to their weak and strong interactions. AB - Figure 3 summarizes the effects of actomyosin binding on the internal and global dynamics of either protein, as discussed in this chapter. These effects depend primarily on the strength of the interaction; which in turn depends on the state of the nucleotide at the myosin active site. When either no nucleotide or ADP is bound, the interaction is strong and the effect on each protein is maximal. When the nucleotide is ATP or ADP.Pi, or the equivalent nonhydrolyzable analogs, the interaction is weak and the effect on molecular dynamics of each protein is minimal. The weaker effects in weak-binding states are not simply the reflection of lower occupancy of binding sites--the molecular models in Fig. 3 illustrate the effects of the formation of the ternary complex, after correction for the free actin and myosin in the system. Thus EPR on myosin (Berger and Thomas 1991; Thomas et al. 1995) and pyrene fluorescence studies on actin (Geeves 1991) have shown that the formation of a ternary complex has a negligible effect on the internal dynamics of both [figure: see text] proteins (left side of Fig. 3, white arrows). As shown by both EPR (Baker et al. 1998; Roopnarine et al. 1998) and phosphorescence (Ramachandran and Thomas 1999), both domains of myosin are dynamically disordered in weak-binding states, and this is essentially unaffected by the formation of the ternary complex (left side of Fig. 3, indicated by disordered myosin domains). The only substantial effect of the formation of the weak interaction that has been reported is the EPR-detected (Ostap and Thomas 1991) restriction of the global dynamics of actin upon weak myosin binding (left column of Fig. 3, gray arrow). The effects of strong actomyosin formation are much more dramatic. While substantial rotational dynamics, both internal and global, exist in both myosin and actin in the presence of ADP or the absence of nucleotides, spin label EPR, pyrene fluorescence, and phosphorescence all show dramatic restrictions in these motions upon formation of the strong ternary complex (right column of Fig. 3). One implication of this is that the weak-to strong transition is accompanied by a disorder-to-order transition in both actin and myosin, and this is itself an excellent candidate for the structural change that produces force (Thomas et al. 1995). Another clear implication is that the crystal structures obtained for isolated myosin and actin are not likely to be reliable representations of structures that exist in ternary complexes of these proteins (Rayment et al. 1993a and 1993b; Dominguez et al. 1998; Houdusse et al. 1999). This is clearly true of the strong-binding states, since the spectroscopic studies indicate consistently that substantial changes occur in both proteins upon strong complex formation. For the weak complexes, the problem is not that complex formation induces large structural changes, but that the structures themselves are dynamically disordered. This is probably why so many different structures have been obtained for myosin S1 with nucleotides bound--each crystal is selecting one of the many different substates represented by the dynamic ensemble. Finally, there is the problem that the structures of actomyosin complexes are probably influenced strongly by their mechanical coupling to muscle protein lattice (Baker at al. 2000). Thus, even if co-crystals of actin and myosin are obtained in the future, an accurate description of the structural changes involved in force generation will require further experiments using site directed spectroscopic probes of both actin and myosin, in order to detect the structural dynamics of these ternary complexes under physiological conditions. PMID- 11892286 TI - Familial hypertrophic cardiomyopathic myosin mutations that affect the actin myosin interaction. PMID- 11892288 TI - Blood bags. PMID- 11892287 TI - Coupling between chemical and mechanical events and conformation of single protein molecules. PMID- 11892289 TI - NICE issues guidance on hips. PMID- 11892290 TI - TAQ: theatre access qualification course. PMID- 11892291 TI - Waste anaesthetic gases. AB - Occupational exposure to inhalation anaesthetics is a controversial subject and an topic for ongoing debate. In this article, Vanessa Pressly reviews the significant research on the subject and details current regulations and guidelines for perioperative practice. PMID- 11892292 TI - Risk for the new or expectant mother working in the perioperative environment. AB - Being pregnant is not an illness, and in the National Health Service, where the majority of the workforce are female, pregnancy should be regarded as part of everyday life. Yet the workplace can damage your health, and that of an unborn child, through hazardous substances such as chemicals, radiation, and anaesthetic gases; through work which involves moving and handling of loads; through stress, excessive hours or shift work; and for perioperative practitioners, through standing for long periods of time (Rogers et al 1999). PMID- 11892293 TI - Call us first--we'll fix it. How a day case unit plays a vital role through teamwork. AB - Clinical effectiveness can be defined as 'the extent to which specific clinical interventions, when deployed in the field for a particular patient or population, do what they are intended to do ... maintain health and secure the greatest possible health gain from the available resources.' (NHS Executive 1996) 3M/NATN Award winner for 1999 Bobbie Lang describes how her Day Case Unit has become the central focus for the co-ordination of such procedures--offering clinical effectiveness for a particular patient group. PMID- 11892294 TI - Nursing at the forefront. AB - The Government sees nurses at the forefront of the drive for improvement in healthcare delivery. In this article, Lord Hunt, the Government spokesperson on nursing, spells out the role that nurses are expected to play in this process. Clinical governance has undoubtedly already been mentioned if not enacted in your Trust, and maybe some of you have applied for the discretionary points now on offer. Whatever the outcome of these and other strategies, the future is going to be different for perioperative nurses, whether we like it or not. Your views, as ever, are welcome.... PMID- 11892295 TI - Reducing patient stress in theatre. Alison Bell Memorial Award. AB - For most patients, admission to hospital for surgery can be very stressful. It is well recognised that stress, created by prolonged anxiety, can produce harmful effects and may even delay a patient's recovery following surgery. This article, submitted for the 1999 Alison Bell award (sponsored by NATN and Regent Medical), looks at key ways to reduce the effects of such stress. PMID- 11892296 TI - Hand washing. AB - What could be more basic than the simple act of washing your hands? Children at junior school are taught to do it. Yet research tends to point to the lack of hand washing as being a major contributor to hospital acquired infection. I have summarised this piece by Elizabeth Pinney which takes us right back to the 18th century--and bang up to date! Reflect on your practices and that of your colleagues before you tell me this is old news. PMID- 11892297 TI - Instrument log, star date ... 2000. PMID- 11892298 TI - Moving towards evidence based care. PMID- 11892299 TI - Latex facts in perioperative practice. PMID- 11892300 TI - Where have all the students gone? PMID- 11892301 TI - Theatre nursing in Finland. AB - As part of the Socrates exchange system three students from the School of Care Sciences, University of Glamorgan visited Finland for a 15-week placement. They were based in two areas Hameenlinna and Kuopio. As part of the exchange I visited the students, did some teaching and observed some of the clinical areas. PMID- 11892302 TI - Hand washing. Simple, cost effective, evidence based ... lip service! AB - Hand washing is so simple that we teach our toddlers to do it. It's so important in our every day lives that facilities for hand washing can be found in every conceivable environment. Scientists, literature and the media give evidence on the benefits and in the art of good techniques. So why, in an environment where the management of infection is a matter of health and safety for all, is hand washing so low on our ever-growing list of priorities? This article examines the legal and moral issues of hand washing in relation to health and safety, concentrating on possible reasons for non-compliance and how these can be addressed. PMID- 11892303 TI - Patient perceptions of day surgery. AB - Having an operation is always a cause for some anxiety, and probably fear for most people. Being told that you don't need to stay in hospital, and that your surgery can be done as a day case with no need for even a one night stay would probably be a comfort. The thought that it will be all over on one day, and that you can go home to your own bed, makes the prospect less daunting. Indeed research has shown that patients prefer day surgery for these and other reasons. Because day surgery means more patients treated at less cost than inpatient surgery, the Government is encouraging the expansion of day surgery services and Trusts are responding. Treating more patients more quickly has got to be good news for the many thousands of patients on waiting lists, but having your operation as a day case should not mean production line treatment. Caring for the personal and meeting individual needs is equally applicable to day case patients as it is to inpatients. This study presents an exploration of some of the actual and potential problems which need to be addressed if day surgery is to become part of the 'first class service' the Government has promised. PMID- 11892304 TI - Nursing. Image, politics and the media. AB - This is the final article in the series on nursing and politics. It examines one person's perspective on the image of nursing as portrayed in the media, and the place of nursing in political debate. The series has presented a variety of views: from political correspondents, from Government or Opposition spokesmen. This final article offers the views of Ed Doolan, a journalist and BBC broadcaster who is also a member of an NHS Trust Board. The article is based on an interview which took place at the BBC Pebble Mill studios in Birmingham in February this year. Ed Doolan has not been paid a fee for the interview; instead, NATN have made a contribution to the Children's Hospital, Birmingham. PMID- 11892305 TI - Back to basics in anaesthesia. AB - This is the last in the Back to Basics series of articles--for now at least. I would like to thank all the authors for their pieces. I've learnt and relearnt from each of them. The completed set of articles will be available as one document in October, and will be launched at Congress--yet another reason to attend! Bye for now. PMID- 11892306 TI - US conference highlights HCV risk. PMID- 11892307 TI - Malnutrition in hospitals. PMID- 11892308 TI - A new breed of workers? PMID- 11892309 TI - Metal diathermy plates. PMID- 11892310 TI - Latex facts in perioperative practice. PMID- 11892311 TI - Realising your assets. PMID- 11892312 TI - New law, new nurses. PMID- 11892313 TI - Critical incident. An ethical dilemma. AB - The purpose of this critical incident analysis is to demonstrate how knowledge of legal, moral and ethical issues has affected my nursing practice. This is a personal journey--an exploration of my own values and beliefs. The piece demonstrates how reviewing the literature has changed my stance on the issues discussed. Consequently, the piece is written in the first person, but references are used to substantiate my findings. PMID- 11892314 TI - Verbal abuse. AB - The Health and Safety Executive includes verbal abuse in its definition of work related violence (HSE 1997). Verbal abuse is 'any remark made to or about a client which may reasonably be perceived to be demeaning, disrespectful, humiliating, intimidating, racist, sexist, homophobic, ageist or blasphemous'. Examples include making sarcastic remarks, using a condescending tone of voice or using excessive and unwanted familiarity (UKCC 1999). PMID- 11892315 TI - Body language--non-verbal cues. AB - Interviews for employment are significant events in peoples lives. Preparing for one and getting through it can be a memorable experience, good or bad. We all know that what we don't say can be as important as what we do say. In this fascinating piece of research Lesley Mason describes how she was able to relate the non-verbal communication of interview candidates to the outcome of the interview. Read on and find out what you were really saying as you sat, squirmed or twiddled your way through that last interview.... PMID- 11892316 TI - The role of chief nursing officer. AB - This is an important time for nurses, midwives and health visitors in the NHS. It is a time of radical change, of real investment and real challenges. I am pleased to take this opportunity to contribute to your journal and to speak directly to perioperative nurses about these issues. I want to talk to you about my own role, the imminent changes to the NHS, and how I see us working together in the future. PMID- 11892318 TI - Pre-op warming reduces infections. PMID- 11892317 TI - From the outside looking in. The surgical approach. AB - This series of articles will draw on Physiology and Anatomy, a homeostatic approach (hereafter referred to as 'the reader'), written by John Clancy and Andrew McVicar. There will also be cross-references to the series of articles entitled 'Homeostasis--the key concept to physiological control' published in the British Journal of Theatre Nursing 1996-98. The articles will include some activities which the readers can carry out to test their understanding and deepen their knowledge base of the anatomy explored in each article. The introductory article will cover the common anatomical structures involved in the 'surgical approach', looking first at the anatomy and then relating it to surgical practice. Homeostasis is the key underlying principle to which the surgical practice will refer. PMID- 11892319 TI - Latex facts in perioperative practice. PMID- 11892320 TI - Where have all the students gone ... again? PMID- 11892321 TI - Life at the Britfor Military Hospital in Kosovo. AB - Life is full of new experiences and on 27 April 2000 a new one occurred for me. As a Nursing Officer in QARANC I was deployed for three months to the Britfor Military Hospital in Pristina, Kosovo to join 22 Field Hospital for my first operational tour. I was to be the only theatre sister in charge of two surgical teams which comprised five theatre technicians, two anaesthetists and two surgeons (one general and one orthopaedic). PMID- 11892322 TI - Confidentiality. The Caldicott Report. AB - You will probably look at the title of this and want to move on to more interesting material, but do hang on for a little bit longer. You may already know about it, but there will be an impact on you and your department as a result of this initiative. Caldicott may just be one of the seminal moments in the development of the NHS; it does have impact and could develop the cause of patients' rights a little bit further. This is a very broad overview, but it does have some helpful hints. PMID- 11892323 TI - The musculo-skeletal system. The spine. AB - This is the second of a series of articles based on Physiology and Anatomy, a homeostatic approach by John Clancy and Andrew McVicar. This article looks at the musculo-skeletal system, with particular emphasis on the spine. It identifies spinal deformities, injuries and maladies, and various types of surgical intervention. PMID- 11892325 TI - Diathermy and pacemakers. PMID- 11892324 TI - Infection control issues in the orthopaedic theatre environment. AB - This article is the First Prize winning entry in the Essay Section of the Alison Bell Memorial Writer's Award 2000, sponsored by NATN and Regent. It is based on an essay submitted for the Evidence Based Practice Module of the post-graduate programme in Health Promotion and Health Studies at Bath Spa University College, Bath. PMID- 11892326 TI - Latex facts in perioperative practice. PMID- 11892328 TI - A risky business. AB - Just when you get comfortable with life--you've got everything under control and you know what you're doing--along comes someone with a new idea with a change attached to it. From the European Courts, through the Government to the Trust Executive and then the Directorates, change seems to be a constant part of life. Julie Sprague is a theatre nurse who saw an opportunity to make a worthwhile change in her area of practice. This is the acceptable face of change, at clinical practice level, done with involvement and co-operation of colleagues. This article describes that change process together with the theories and rationale which support it. PMID- 11892327 TI - The use of touch for a patient experiencing laryngospasm. AB - This article is based on a course work assignment for the Welsh National Board Certificate Level Module in Operating Department Practice (Recovery Branch). The article uses a critical incident reflective approach to explore the issues of using touch in caring for a patient, experiencing laryngospasm following a general anaesthetic. A literature review will explain the mechanisms of laryngospasm and will discuss the use of different forms of touch. Conclusions and recommendations will be drawn from the issues discussed. PMID- 11892329 TI - Bone cement mixing. Theatre staff's views and opinions. AB - Acrylic bone cement has not substantially changed since it was first introduced more than 30 years ago, however the method by which it is mixed has evolved greatly (Dunne 1996). PMID- 11892330 TI - Oscillometric blood pressure monitors. AB - The mercury sphygmomanometer has been used to measure blood pressure for over a century (O'Brien 2000, Smith 2000), but due to the hazards associated with mercury spillage, there is increasing reliance on other methods, such as automatic oscillometric blood pressure monitors and aneroid sphygmomanometers. Within the theatre and recovery areas, the oscillometric method appears to be most common (Ramsey 1991). PMID- 11892331 TI - Protect yourself! PMID- 11892332 TI - HIV on increase in UK. PMID- 11892333 TI - Sterile items were still safe to use even if they had passed the stamped expiry date. PMID- 11892334 TI - Latex facts in perioperative practice. PMID- 11892335 TI - Half empty, half full, half cock? PMID- 11892336 TI - The preparation of bone cement. AB - The hip joint is subjected to large, repetitive loads. It is therefore clear that the bone cement, which allows the transfer of load across the new joint, must be able to withstand the everyday loads that it will be subjected to. Improving the mechanical properties of the cement to withstand high stresses, fatigue and creep loading will reduce the chances of failure, ultimately increasing the longevity of the joint replacement. To date, work in this area has concentrated on improving the mechanical properties of bone cement through improved bone cement mixing techniques. In the next issue we will be covering the effect that the design of the mixer and vacuum mixing has on improving the mechanical properties, such as the strength, fatigue and creep resistance, of the bone cement. PMID- 11892337 TI - Creutzfeldt-Jakob disease. The story so far. AB - CJD continues to be an increasing problem for perioperative practitioners to be aware of. Maureen Dyke reviews the implications for perioperative practice. PMID- 11892338 TI - Professional vs commercial. Can they be reconciled? AB - This is the first in a series of articles looking at the challenges facing everyone who works in healthcare, whether in the public or private sector. In this article, Fiona Westwood addresses the question: Professional vs Commercial: can they be reconciled? Other questions that will be discussed over the coming months are: What is effective management? What forms of leadership and communication really work? Are we truly patient-focussed? How well do we use our resources? And, is it possible to manage professionals? PMID- 11892340 TI - Of vision, virtue, valour and victory. PMID- 11892339 TI - Interprofessional working and learning in a UK operating department. AB - This study explores the issues of working and training together for nurses and operating department assistants and practitioners (ODAs/ODPs), based on experiences in an operating department. Interviews and focus groups with nurses, ODAs/ODPs and medical staff were the means of collecting data. PMID- 11892341 TI - Online education demo. How to get started! AB - The purpose of this article is to provide a brief tutorial on how to use the online education demo that is on the NATN website. PMID- 11892342 TI - An ethical dilemma. PMID- 11892343 TI - We still use Disposable shoe covers for visitors to the department. PMID- 11892345 TI - AIDS on the rise in the UK. PMID- 11892344 TI - Latex facts in perioperative practice. PMID- 11892346 TI - Manna from heaven. Welcome to the information age! PMID- 11892347 TI - Temperature and its effect on bone cement. AB - Bone cement mixing is often seen as a simple procedure, but the skill of the perioperative practitioner in mixing the cement may ultimately be the difference between the success and failure of a hip or knee joint replacement. Klaus-Dieter Kuhn (2000) states: 'The nurses (together with the surgeon) have enormous influence on the quality of the cement dough produced; in the end, this will considerably influence the clinical long-term result of a cemented hip, acetabulum or cemented knee.' (Bone Cements). This process doesn't happen by default. It is the result of good education, supported by sound evidence and backed up by practical experience. This series of articles has explained every aspect of cement management, covering technique, temperature, timing, mixing equipment, the importance of reproducible cement quality and health and safety issues. PMID- 11892348 TI - Annualised hours. PMID- 11892349 TI - First assistant activities. AB - A discussion of some of the issues surrounding the first assistant role has been ongoing in perioperative journals since around 1995. The crux of this discussion is the concern that theatre nurses who take on first assistant duties will encounter problems with professional accountability and liability. It is also proposed that theatre nurses wishing to act as first assistants require additional training. However, much of this debate is influenced by an assumption that has been accepted, apparently unchallenged, by some theatre nurses. This assumption is that first assistant activities constitute an extended role. PMID- 11892350 TI - Gastroentological investigations. AB - In this fourth article in the Concepts in Anatomy series, Anne-Marie Ramsay joins John Clancy and Andrew McVicar in examining the upper and lower gastrointestinal tract and identifies some common homeostatic imbalances. Diagnostic tests for detecting abnormalities are described, highlighting the importance of the nurse's role in caring for the patient throughout these procedures. This series is based on Physiology and Anatomy, a homeostatic approach, 2nd ed, John Clancy and Andrew McVicar (eds), Edward Arnold, London, 1995, currently in print. PMID- 11892351 TI - Leadership and communication. What really works? AB - In her previous article, Fiona Westwood looked at effective management and what this means in practice in our organisations. This article develops these themes by considering the role of leadership and the importance of open and honest communications. PMID- 11892352 TI - Occupationally acquired HCV--the human story. PMID- 11892353 TI - Within our department, it is current practice to allow staff to leave the perioperative environment wearing their scrub attire. PMID- 11892355 TI - Waving not drowning! PMID- 11892354 TI - Childcare support. PMID- 11892356 TI - Are we truly patient focussed? AB - The end user of all healthcare activity is the patient. But how do we know whether we are providing a good service? Do we take time to listen to our patients, or do we simply hide behind our professional title and pass the buck to someone else? In this, the fourth article of our series, Fiona Westwood asks some challenging questions. The answers may indicate that some changes need to be made. PMID- 11892357 TI - Latex allergy within the perioperative area. AB - Latex allergy is a subject which is causing concern both amongst healthcare workers and in the Department of Health, with reports that significant numbers of NHS staff may have latex hypersensitivity. With this informative article, Janet Cox joins John Clancy and Andrew McVicar to explain the mechanisms that underlie latex hypersensitivity and to make recommendations about how hospitals can deal more effectively with it. The article is based on the text, Physiology and Anatomy, a homeostatic approach, 2nd ed, John Clancy and Andrew McVicar, Arnold, London (in print September 2001). PMID- 11892358 TI - Staff rostering in the operating department. AB - The smooth running of the operating department is often facilitated by the goodwill of the staff who agree to stay behind when cases overrun, or work through lunch and coffee breaks. Many perioperative staff also cover on-calls and stand-bys, causing disruption to their social and family lives. These demands may make working in the operating department harder for some staff. With reference to current shortages of perioperative staff and staff retention, Judith Tanner and Gloria Bailey discuss some suggestions for being flexible with the off duty rota which may give staff more control and more choices over when they work. PMID- 11892359 TI - [Myocardial regulatory proteins in children with congenital heart defects]. AB - The purpose of this study was to compare the protein profiling of the regulatory proteins. The samples of myocardial tissue were obtained during surgical intervention from children (age 8.2 +/- 1.8 years) operated for different types of congenital heart diseases. The arterial oxygen saturation was 95.5 +/- 0.07% in normoxaemic patients (ventricular and atrial septal defects), resp. 76.9 +/- 2.1% in hypoxaemic patients (tetralogy of Fallot). The both cytosolic and myofibrillar fractions of cardiac troponin T (cTnT) were isolated by stepwise extraction (4) from the both right ventricular and right atrial musculature. The concentration of proteins was measured using Coomasie Plus Protein Reagent Kit (Pierce). After isolation, one-dimensional SDS polyacrylamide gel electrophoresis was carried out according to a modified version of the method of Laemmli (1970), using a 12% separating gel and a 4% stacking gel. In some cases after SDS-PAGE, proteins were electroblotted onto nitrocellulose membrane for immunoblotting and analysed using the JLT-12 monoclonal antibodies (Sigma Chemicals). The cytosolic pool of cTnT (measured by commercial kit Elecsys Troponin T STAT Immunoassay- Roche) represents about 12.5%, the myofibrillar pool of cTnT was about 87.5%; hypoxaemia did not affect this proportion. PMID- 11892360 TI - [Changes in the quality of life after cataract surgery]. AB - The questionnaire VF-14 is one of the most frequently used methods for exploration of patients' subjective evaluation of cataract surgery outcomes. In this study, the answers of 349 patients' were evaluated. These respondents completed the questionnaires before cataract surgery and again after 6 months. When the postoperative questionnaires were filled-in, 196 respondents had one eye operated on (group A), and 153 had both eyes operated on (group B). All cataract surgeries were performed by phacoemulsification. The complication--rupture of the posterior capsula--occured in 0.8%. In 26.7% patients an accompanying secondary eye disease was found. The average VF-14 index was 63.5 (SD 21.32) in group A and 59.4 (SD 22.62) in group B. After the surgery the VF-14 index increased significantly--in group A was the average 80.6 (SD 20.47) and in group B even higher 87.2 (SD 16.14). The change of VF-14 index prior to and after the operation was proven to be statistically significant both in group A and in group B (p < 0.001). The improvement of the VF-14 index was found in group A in 80.6% and in group B in 88.9% of respondents. The patients' satisfaction with their vision after cataract surgery was highly increased. The outcomes of the questionnaire study document the high success rate of cataract surgery for patients. PMID- 11892361 TI - [New findings in monitoring health status of welders and grinders of stainless steel]. AB - During the occupational preventive care check up we investigated a group of 20 stainless steel welders and grinders in the factory producing technology for chemical industry. Results have been compared with 21 healthy persons--blood donors. In the group exposed there have not been discovered any marked deviations in either their health status, or in the CBC and biochemical screening results, though there had been found substantially increased chromium and nickel concentrations, mainly in grinders. Also the levels of polycyclic aromatic hydrocarbons were in some cases even higher than on the busy city crossing. The chromosome aberrations investigations proved to be very sensitive and confirmed that employees of the followed up factory are exposed to an increased genotoxic risk. PMID- 11892363 TI - [Assessment and development of occupational diseases in health care workers in the eastern regions of the Czech Republic 1986-2000]. AB - The present authors have summarized their experience of assessing occupational diseases in the health-service workers from the catchment area of the Department of Occupational Diseases of the Faculty of Medicine and the Faculty Hospital in Hradec Kralove reported in the years 1986-2000. It is of great importance that, in spite of a decrease in occupational diseases in the Czech Republic as a whole, in health service workers occupational diseases still take a prominent place. In the Region of East Bohemia health services permanently occupy the first place, followed by agriculture and metalworking industry, which alternate. A positive finding in the number of viral hepatitides B has significantly decreases. On the other hand, the number of patients suffering from scabies and, above all, allergic diseases, in which contact allergic eczemata due to hypersensitivity to latex contained in protective gloves inequivocally prevail, is on the increase. Though there is no increase in the number of patients with tuberculosis, there is an important finding that the disease affects mainly persons younger than 35 years. PMID- 11892362 TI - [Plasma levels of troponin T in rabbits after administration of new antineoplastic agents (Oracin and Dimefluron) and daunorubicin]. AB - Recently, cardiac troponin T (cTnT) has been shown to be a sensitive marker of anthracycline-induced cardiomyopathy. In our study, the cardiotoxicity of repeated i.v. administration (once a week, 10 administrations) of daunorubicin combined with new antineoplastic drugs (with mild side-effects) were followed in two groups of rabbits: 1) Dimefluron (3,9-dimethoxybenfluron hydrochloride-12 mg/kg) + daunorubicin (3 mg/kg), 2) Oracin (6-[2-(2-hydroxyethyl)aminoethyl]-5,11 dioxo-5,6-dihydro-11H- indeno[1,2c]isoquinoline hydrochloride--10 mg/kg) + daunorubicin (3 mg/kg) and compared with the control group (saline--1 ml/kg) and the group with experimentally induced cardiomyopathy (daunorubicin--3 mg/kg). The concentration of cTnT in heparinized plasma samples was measured using commercial kit (Roche). In the control group, plasma levels of cTnT were always within the physiological range (i.e. lower than 0.1 microgram/l) during the experiment. During the development of daunorubicin-induced cardiomyopathy, after the eighth administration of drug, cTnT was significantly higher (0.31 +/- 0.11 microgram/l) in animals with premature deaths compared with the rest of the group (0.04 +/- 0.03 microgram/l). The animals with pathological values of cTnT were at higher risk of premature deaths (P = 0.0006). The combination of daunorubicin either with Oracin or with Dimefluron caused neither significant changes of cTnT levels nor significant deterioration of other followed-up parameters (especially, functional and toxicological parameters). Similarly to the daunorubicin group, the animals with pathological levels of cTnT after the eighth administration of antineoplastic drugs were at higher risk of premature death (P = 0.025). Our results show that the plasma concentration of cardiac troponin T could be a suitable predictive marker of cardiotoxicity of antineoplastic drugs. PMID- 11892364 TI - [History of chest percussion]. AB - Although percussion of the abdomen was already known to the Greek physician Galenos (2nd century A. D.) who used it to distinguish between ascites and meteorism, the Viennese physician Leopold Auenbrugger (1722-1809) started to use, the percussion of the chest as a diagnostic tool. In 1761 he published his experience in a treatise called "New Method for Detecting Hidden Ailments of the Chest by Percussion of the Thorax" (Inventum novum ex percussione thoracis hummani ut signo abstruso interni pectoris morbos detergendi). However, this method was introduced into practice only 50 years later by Jean Nicolas Corvisart, who translated Auenbrugger's book in 1808 into French. The famous Vinnese internist of Czech origin Joseph Skoda (1805-1881) set the teaching about percussion and auscultation on a firm physical basis. Skoda confronted the physical findings with dissection materials in close cooperation with the renowned Vinnese pathologist Karl Rokitansky (1804-1878), who was born in Hradec Kralove. The Medical School in Prague became famous for its excellent command of methods based on physical examination and surpassed even the Viennese School. PMID- 11892365 TI - [History of health care, hospitals and medical school in Hradec Kralove]. PMID- 11892366 TI - [Cytoimmunofluorometry in the diagnosis of MALT lymphoma of the gastrointestinal tract]. AB - MALT lymphoma is raised from lymphoid cells of mucosa associated lymphoid tissues in a long-term, multistep process of the accumulation of genomic damage very often under the influence of chronic (e.g. Helicobacter pylori) infection. Cytoimmunofluorometric analysis of cell suspensions obtained from mechanically disintegrated bioptic samples of gut mucosa is a very useful approach to detect the presence of MALT lymphoma cells. MALT lymphoma cells are in a great majority of B cell origin. Malignant cells are characterized by the typical characteristic SS parameter, strong expression of CD20 and CD79b molecules, and by the absence of CD5 and CD23 molecules. The ultimate evidence of clonality is immunoglobulin light chains either kappa or lambda expression determination. PMID- 11892367 TI - The relationship of lip strength and lip sealing in MFT. AB - The purpose of this study was to explore the relationship between lip sealing and lip power, and the effect of button pull exercise on lip posture and lip power. 91 patients who had barely acquired lip sealing had received button pull exercise. They were evaluated for lip power and lip seal before and after oral myofunctional treatment. In spite of contrary postures of lip between the Button Pull Group and the Non-Button Pull Group no significant difference for lip strength was found at the first examination. The lip strength of the Button Pull Group had increased twice as much after a half-year and decreased thereafter. 25% of the Button Pull Group acquired complete lip sealing after the treatment, 41% did incompletely and 31% did not change. PMID- 11892368 TI - Rationale for including orofacial myofunctional therapy in university training programs. AB - This article provides information on the need felt by speech-language pathologists for training in orofacial myofunctional phenomena. Results of a survey indicate 97.7% of respondents felt training is necessary, while only 7.9% felt their training was adequate. ASHA position statements regarding orofacial myology are reviewed. ASHA and IAOM suggested competencies are also included. PMID- 11892369 TI - Prevalence of adapted swallowing in a population of school children. AB - Adapted swallowing (AS) and its effects on dentofacial balance are frequently observed in children with orofacial myofunctional alterations. The preliminary objective of this research was to identify the frequency of cases of adapted swallowing in a population of school children, and targeting early treatment to prevent morphofunctional alteration of the face and its consequences on dentofacial harmony. One hundred school children between seven and nine years old attending the Children and Adolescents' Supporting Program (PRODECAD) of the State University of Campinas (UNICAMP) Brazil were selected at random and examined. Orofacial myofunctional assessments were carried out through standardized protocol. Results indicated the prevalence of adapted swallowing associated with resting posture alterations of dentofacial structures and mastication with dentofacial disharmony was 57%. Adapted swallowing without alteration of dentofacial form was observed in 19% of cases, and 24% of cases had a swallowing pattern within normal limits. The high prevalence of alterations suggests the need for early professional intervention. PMID- 11892370 TI - Orofacial myofunctional disorders related to malocclusion. AB - The purpose of this article is to enhance awareness about different pathologies that can be minimized or alleviated simultaneously. The author writes about the assessment, the etiologies, the differential diagnosis and the most important interdisciplinary team. PMID- 11892371 TI - Functional outcomes of orofacial myofunctional therapy in children with cerebral palsy. AB - Though some anecdotal evidence supports the efficacy of orofacial myofunctional therapy in cerebral palsy and other disorders, controlled studies are very scant. This study was undertaken to examine the efficacy of orofacial myofunctional therapy in sixteen children diagnosed with spastic cerebral palsy. Following baseline measures, all children participated in a four-month therapy program, consisting of training the tongue, lips, and jaw muscles for adequate posturing and functioning. Post-therapy measures indicated significant improvement in functioning of lips, tongue, and jaw. Speech intelligibility of words also improved significantly as measured by two judges using a five-point rating scale. A significant correlation was found between tongue functioning and improvement in speech intelligibility; however no significant correlation was obtained between functioning of lips/jaw and speech intelligibility. Clinical implications regarding use of orofacial myofunctional therapy with cerebral palsied children are discussed. PMID- 11892372 TI - Managing foot ulcers in patients with diabetes. AB - Over 1 million people in the UK have diabetes mellitus, of whom around 7-10% will, at some point, develop a foot ulcer. These ulcers, which in severe cases extend to muscle and bone, are prone to infection and can lead to gangrene and may necessitate amputation. Around 85% of amputations in people with diabetes are preceded by persistent foot ulcers. Here, we review the management of such ulcers. PMID- 11892373 TI - [symbol: see text]Quinupristin + dalfopristin for infections. AB - Quinupristin + dalfopristin* (Synercid -- Aventis Pharma Ltd) is a new combination antibacterial product licensed for treating patients with a variety of Gram-positive infections. Here we assess its place in clinical practice. PMID- 11892374 TI - Chemotherapy and non-small-cell lung cancer. AB - Around 34,000 people in the UK die from lung cancer each year. Over 75% of patients with the disease have non-small-cell lung cancer (NSCLC). Here, we discuss the role of chemotherapy in the management of NSCLC, a treatment that some medical oncologists estimate would benefit 50% of patients with NSCLC, but which is given to fewer than 10%. PMID- 11892375 TI - Price survey. Endoscopic devices face price hikes. PMID- 11892376 TI - CHeS nears agreement on med-surg terminology. PMID- 11892377 TI - To find savings, look at the big picture. AB - Taking a scientific approach to supply cost management is a lot of work, but the effort can net big savings, as a California hospital found last year. Hoag Memorial Hospital Presbyterian saved $2 million on supplies and services in 2001 and expects to continue saving that amount annually for the next couple of years. PMID- 11892379 TI - Novation cracking down on bar code use. PMID- 11892378 TI - Higher revenues, lower prices seen in 2002. AB - Economic growth will pick up in 2002, with higher revenues for most industries, while prices will remain stable or continue to drop. That is the prediction of the Institute for Supply Management (ISM), formerly National Assn. of Purchasing Management. PMID- 11892380 TI - Surface enhancement to optimize the success of dental implants. Interview by Arun K. Garg. PMID- 11892381 TI - Universal newborn hearing screening: a goal being achieved in Hawaii. AB - This article describes the importance of early identification of hearing loss in newborns, the current status of newborn hearing screening in the United States, and the leadership that Hawaii has contributed to that effort. Described are events that may help the nation reach the Year 2010 Health Goals for newborn hearing screening, identification, and intervention. PMID- 11892382 TI - The role of geriatrics and gero-psychiatry in medical education. PMID- 11892383 TI - Searching for the causes of gastric cancer. PMID- 11892384 TI - [Electrophysical effects in combined treatment of neurosensory hypoacusis]. AB - The authors consider different methods of electrobiophysical impacts on the body in the treatment of neurosensory hypoacusis: laser beam, laser puncture, electrostimulation, magnetotherapy, magnetolasertherapy, electrophoresis, etc. These methods find more and more intensive application in modern medicine. Further success of physiotherapy for neurosensory hypoacusis depends on adequate knowledge about mechanisms of action of each physical method used and introduction of novel techniques. PMID- 11892385 TI - [Current aspects of endolaryngeal and endotracheal microsurgery in children]. AB - Achievements of today's laryngeal and tracheal surgery came for the most part from novel optic techniques and anesthesia. All the operations were performed with application of the microscope or endoscopic devices. Gas exchange was provided differently: using volume, injection and high frequency artificial ventilation of the lungs. An individual approach to gas exchange technique in respiratory obstruction allowed to avoid emergency tracheotomy. A laryngeal mask was used in children for air delivery in fibrotracheobronchoscopy. Removal of tumors, dissection of laryngeal scars, trachea and bronchi were made by means of ultrasound and laser equipment. Interventions are considered for each condition. PMID- 11892386 TI - [Immune status of patients with chronic tonsillitis before and after tonsillectomy]. AB - Immune status was examined before and after tonsillectomy in 35 patients with various forms of chronic tonsillitis (CT). Measurements were made of main lymphocyte populations, IgA, IgG, IgM levels, functional activity of neutrophils, titers of antibodies to opportunistic microorganisms of the upper airways. It is shown that tonsillectomy produces positive changes in some immunological indices: serum IgG normalized, titers of antibodies to Haemophillus influenzae, Streptococcus pneumoniae and streptolysin-0 lowered, initially low count of B lymphocytes, natural killers, T-helpers rose, neutrophil chemiluminescence normalized. PMID- 11892387 TI - [Radionuclide visualization of reparative osteogenesis after obliteration of the paranasal sinuses with bio-compositional implants]. AB - 18 patients with chronic recurrent inflammation and traumatic defects of the paranasal sinuses have undergone obliteration of the sinuses with biocompositional materials (hydroxyapatite and colapol) and plastic reconstruction of the bone defects with metal ceramics. Control of the repair processes with emission computed tomography registered accumulation of radiopharmaceuticals (RP) in the obliteration zone a week after the operation. Four-six weeks later RP homogeneously filled the obliteration zone. Six-twelve months after the obliteration, metabolism in the implant is close to that in the intact tissue. It is concluded that emission computed tomography can follow up metabolic changes in the implant and assess efficiency of the operation. PMID- 11892388 TI - [Endonasal micro-dacryocystorhinostomy in nasolacrimal obstruction]. AB - Disorders in lacrimal passage are often caused by anatomotopographic peculiarities of the nasolacrimal tract. Surgical treatments of such patients are reviewed. The authors made endoscopic endonasal dacryocystorhinostomy in 34 patients with good results. How to achieve good functional outcomes is described. PMID- 11892389 TI - [Treatment of frontal sinus inflammation]. AB - The analysis of urgent treatment of frontal sinus inflammation from 1985 to 1999 and 6 months of 2000 made in ENT clinic of Izhevsk shows advantages of conservative therapy (trepanation puncture and frontal sinus probing) in diagnosis and treatment of frontal sinus inflammation. Literature data on safety of such manipulations is confirmed. PMID- 11892390 TI - [Analysis of nasal and paranasal cancer in the Rostov region for 35 years: problems of early diagnosis]. AB - The article contains information on 280 patients with nasal and paranasal cancer of epithelial (85%) and non-epithelial genesis (15%); on diagnosis, spread of the primary tumor, regional and distant metastases; principles of patients grouping according to TNM staging; analysis of causes of late diagnosis and misdiagnosis. PMID- 11892391 TI - [Effectiveness of combined homeopathic drug "Faryngomed" in the treatment of upper respiratory tract diseases]. PMID- 11892392 TI - [Algorithm of examination and treatment of patient with neurosensory hypoacusis]. PMID- 11892393 TI - [Combined orbital and intracranial complication in a female patient with acute rhinosinusitis]. PMID- 11892394 TI - [ENT lesions in children with AIDS]. AB - The number of AIDS patients in Cambodia is on the increase. The examination for AIDS covered 50,000 children and 23,000 adults. HIV-seropositive results were obtained in 1800 and 1250 children and adults, respectively. ENT lesions in children with AIDS are characterized. PMID- 11892395 TI - [A giant osteoma of ethmoidal labyrinth with invasion into the retrobulbar space]. PMID- 11892396 TI - [A rare combination and location of multiple intracranial and orbital complications of acute purulent hemi-sinusitis]. PMID- 11892397 TI - [Laryngeal malacoplakia]. PMID- 11892398 TI - [Tracheal injury in a child]. PMID- 11892399 TI - [A case of bilateral deafness after combined anesthesia]. PMID- 11892400 TI - [Surgical approaches to the treatment of patients with chronic pyohyperplastic rhinosinusitis]. PMID- 11892401 TI - [Late complications of adenotomy]. PMID- 11892402 TI - [Specific features of antibacterial therapy in children with acute otitis media]. PMID- 11892403 TI - [Interrelation between allergic rhinitis and bronchial asthma (review of literature)]. PMID- 11892404 TI - [Cough]. PMID- 11892405 TI - [Some characteristics of optokinetic nystagmus in patients with unilateral vestibular neuronitis]. AB - Optokinetic reflex was studied in 20 patients aged 20-58 years with vestibular neuronitis. In 16 decompensated patients the direction of the optokinetic nystagmus was inversed. This inversion disappeared in development of the compensation. Mean values of the amplitude and speed of the slow phase of optokinetic nystagmus are presented for patients with vestibular neuronitis and 20 healthy subjects in mono- and binocular optokinetic stimulation of various intensity. PMID- 11892406 TI - Network's tools help at-risk providers save millions. PMID- 11892407 TI - California plans collaborate on 'pay for performance' metrics to boost HMO quality. PMID- 11892408 TI - Market analysis documents HMO premium increases by region. PMID- 11892409 TI - State asks court review of Medicaid cap rate decision. PMID- 11892410 TI - Medicare+Choice on the ropes as plan exodus continues. PMID- 11892411 TI - Research attempts to determine best predictor of future high-cost cases. PMID- 11892412 TI - [Course of isocyanate-induced asthma in relation to exposure cessation: longitudinal study of 50 subjects]. AB - Isocyanates, particularly Toluene Diisocyanate (TDI), have been the most common cause of occupational asthma for decades. Fifty workers with a diagnosis of isocyanate-induced asthma were followed up for a mean period of 8.4 years. Pulmonary function testing for FVC and FEV1, allergological tests, nonspecific bronchial challenge to methacholine and specific bronchial challenge to isocyanates were performed at the time of diagnosis and during follow-up. Data on symptoms and drug response were rated on a scale. Patients were subdivided into two groups based on persistence of (A; 13 patients) or removal from (B; 37 patients) isocyanate exposure. There were no significant differences in any characteristic between the two groups at the time of diagnosis. At follow-up, group B subjects showed significant improvement in symptoms, consequent reduction in use of medications and increase in PD20. None worsened, sixteen (43.3%) remained stable, twelve (32.4%) improved and nine (24.3%) became asymptomatic. The latter subjects were generally younger, they had shorter duration of symptoms and exposure to the agents, and longer removal from exposure, although none of these differences was significant. In 67% of cases they had an immediate reaction to TDI challenge. By contrast, the condition of group A patients deteriorated significantly during the follow-up period in terms of symptoms, pulmonary function parameters, PD20 and use of medications. Overall, seven subjects (53.8%) worsened during follow-up and none improved. Group B subjects were further divided into subgroups B1 and B2 based on duration of removal from exposure: B1 < 10 years, B2 > or = 10 years. There were no significant differences between the two subgroups, even though PD20 was higher in B2. The present study confirms that early diagnosis and immediate removal from exposure are crucial, though not always sufficient, for a favourable evolution of the disease. PMID- 11892413 TI - [The Italian version of "OREGE" (Outil de Reperage et d'Evaluation des Gestes) of the INRS (Institut national de recherche et de securite) for the assessment of musculoskeletal disorders of the upper limb]. AB - The upper extremity work-related musculoskeletal disorders (UEWMSDs) are a heterogeneous group of disorders not yet standardised mainly epidemiological criteria for case definition. They are multifactorial, often they are work related even if sometimes they show an individual origin. In recent years they show a rapid increase but it's worth noting that this trend is also affected by a more widespread and easy recognition as work related diseases. There are many ergonomic analysis tools, currently available, that claim to accurately measure variables associated with UEWMSDs. They are essentially based on biomechanical, epidemiological and physiological approaches and identify work activities at risk of developing: OSHA's checklist, Strain Index, OCRA Index, ACGIH (Hand Activity Level). A method for the study of musculoskeletal disorders of the upper limb has been proposed by French INRS (National de Recherche et de Securite). It is defined as a project based on ergonomics applied to occupational medicine and it includes: (1) OSHA's checklist as a screening tool; (2) MSDs questionnaire for standardised record of symptoms and of worker's opinions (3) OREGE, (Outil de Reperage et d'Evaluation des Gestes) a exhaustive evaluation tool to be used by ergonomics-trained personnel, aimed to identify risk factors to be considered for preventive and corrective actions. OREGE includes: force evaluation through Latko's scale (which take into account: weight of objects and tools, kind of hold, pressure, vibration, temperature, use of gloves), articular position analysis, repetition analysis, synthesis of the different biomechanical risk factors, calculation of an index of risk. The authors have considered of interest to provide occupational physicians and ergonomic professionals with an Italian version of OREGE. The expected results are: a critical review of the method, a comparison with the other most known evaluation methods and the selection of the best method for specific work activity. PMID- 11892414 TI - [Occupational exposure to formaldehyde at a service of pathologic anatomy]. AB - OBJECTIVE: Aim of the present study is to evaluate the air pollution produced by formaldehyde in pathological anatomy. METHODS: This study was made with instrumental approach based on environmental evaluation of 10% formaldehyde used in pathological anatomy, by an infrared gas analyser (Bruel & Kjaer), and clinical approach of pathological anatomy personnel. RESULTS: The final result is not very comforting because we found values of formaldehyde during specific activities which exeeded the current limits proposed by industrial hygienist, infact we found in a different settings 1.81 ppm, 3.78 ppm, 8.3.05 ppm. The personnel exposed reported subjective symptoms as reactive airway symptoms, headache, skin problems. CONCLUSIONS: To reduce air pollution we have indicated technical precautions as forced ventilation which is a major engineering control for reducing risk from chemical agents, use of personal protective equipment (PPE) as last resort for protection, behavioral rules and health surveillance. PMID- 11892415 TI - [Environmental and biologic monitoring of atrazine exposure at a formulating plant]. AB - Atrazine exposure was evaluated in three workers engaged in technical Atrazine dust formulating and bagging processes, by environmental and biological monitoring. We found an Atrazine concentration in breathing area of 31.2 micrograms/m3 (SD +/- 9.9); actual inhalatory absorption was 20.7 micrograms/workshift. Cutaneous amount was 6 mg per workshift and the largest fraction was found on hands. Supposing that only 3.5% of the applied was absorbed, 239.7 micrograms of Atrazine were taken up by skin per workshift. The dealkylated metabolites and parent compound excreted in urine, as a whole, accounted for 71.09% of the presumed total dose absorbed. The spectrum or the urinary Atrazine metabolites comprised bi-dealkylated (58%), deethylated (30.7%), deisopropylated (7.7%) and unmodified Atrazine (3.6%). About 31% of the amount is excreted during the workshift itself. These findings allow to evaluate the magnitude of the real doses absorbed, keeping into account the effect of protective means. PMID- 11892416 TI - Effects of noise on functional cardiovascular parameters: a follow-up study. AB - The aim of this follow-up design study is to give a contribution to the knowledge of the controversial relations between noise and functional cardiovascular parameters. The study population consisted of 757 male employees working for 5 different firms in the oil chemical area (refining and distribution of fuel) with different qualifications--345 workers, 212 drivers, 200 clerks--and thus exposed to 3 different levels of noise (85-90 dBA, 80-85 dBA and < 80 dBA respectively). Heart rate, systolic and diastolic blood pressure were monitored 3 days a week at the beginning of the working shift, for a period of 12 years. The analysis of the data obtained indicates that all the 3 parameters under observation tend to increase progressively with the age and working age of the subjects. In the comparison between the 3 levels of exposure, heart rate results not affected by the exposure to noise; systolic blood pressure results sensitive only for the discrimination of the effects due to exposure, showing a statistically significant difference only between clerks on one side and workers and drivers on the other; diastolic blood pressure is highly influenced by the level of exposure to noise, showing significantly different slopes for the 3 groups. In conclusion, blood pressure is a parameter definitely more reliable for the correlation to the level of exposure, while heart rate cannot be considered a marker of exposure to noise. PMID- 11892417 TI - [Risk perception and self-assessment of exposure to antineoplastic agents in a group of nurses and pharmacists]. AB - Many antineoplastic drugs have shown to be carcinogenic, teratogenic and mutagenic to humans and exposure and absorption have been showed to occur during the preparation and administration in health care practise. More recently, in many countries, national health authorities concern has been focused on promoting actions aimed to protect health of the personnel handling these drugs. The present study reports the results of a survey carried out through a questionnaire among the pharmacists and nurses working in the health care settings located in the province of Rome (Italy), with the aim to survey: occupational risk perception of health care workers; their confidence in the safety measures adopted in the work practice; their knowledge of risk factor and the kind of training received regarding these specific agents. Among the respondents, most of the pharmacists (80%) and nurses (90.4%) show a high concern about antineoplastic drugs dangerousness. At the same time, they state not to have a satisfactory level of knowledge regarding the risk factor and, also that their main information source on occupational risk connected to antineoplastic drugs exposure is not the health care structure, where they are employed. Besides, they do not show a high level of confidence in the safety measures adopted in the health care structures with the aim to protect workers' health. The study results suggest the necessity to promote the development of preventive actions aimed to minimise occupational exposure to these substances through the implementation of procedures and controls in the hospitals, as well as the use of work practice and protective equipment, reinforced by workers education and training. PMID- 11892418 TI - [Risk of exposure to UV radiation at biochemical laboratories]. AB - In biological and chemical laboratories UV radiation is largely used, particularly in photo biological and photochemical studies, with the objective to sterilize or to visualize the samples in different technical analysis. In this work we present the results of the evaluation of UV exposition levels and the hazards for researchers and students, that the results shows elevated if they don't use correct laboratory procedures and protective wear. PMID- 11892419 TI - Planning the provision of outpatient services in a rehabilitation clinic. AB - OBJECTIVE: This study had two aims: (1) to plan the provision of outpatient services in a rehabilitation clinic and increase the efficiency of these services; (2) to identify and evaluate criteria of good clinical practice in outpatient rehabilitation activities in order to avoid transforming the therapy into an assembly line in which the number of services is considered more important than the quality of services delivered. DESIGN: The authors identified the sequence of activities carried out for each of the services requested and monitored the number of appointments and treatments performed in two four month periods. As part of the process of increasing delivery efficiency and recording services provided, the authors created a computerised database for processing hospital charges. MAIN MEASURES: Quantitative comparison of services delivered during the 4 month sample periods in 1998 and 1999 and evaluation of compliance with identified indicators of good practice related to a series of rehabilitation outpatient therapies, divided into instrumental therapies and manual therapies. RESULTS: There was an increase in the number of appointments and services without an increase in the number of staff or clinics. In particular, the database introduced in March 1999 contributed to a further improvement with 692 appointments and 7517 therapies. Overall quality of practice was good, although privacy needs to be increased. CONCLUSION: The use of quality control instruments improved staff awareness of good clinical practice and highlighted some aspects which could be improved. The efficiency of the outpatient rehabilitation clinic increased. PMID- 11892420 TI - [Baropodometric walking analysis in healthy elderly and in arthritic patients before and after knee prosthesis implantation]. AB - Standing and walking are complex activities which require integral skeletal muscular and central nervous systems. Body is usually, evenly distributed between both the lower limbs with 40% on the ball of the foot and 60% on the heel, therefore, the body's centre of gravity falls between the feet in correspondence to Chopart's articulation. Some diseases can influence standing and walking including cardiovascular diseases (chronic edema, claudication, cardiopathies etc.) neurosensorial ones (cataract, Meniere's disease, Parkinson's disease etc.) and orthopedic ones (kyphosis, scoliosis, hallux valgus, metatarsalgia, osteoarthritis etc.). In this study arthritis was considered the main cause of changes in posture and deambulation. An electric baropodometer with a modular platform 240 cm long and 40 cm wide was used which provided the pressure information for each in three distinct phases: static, dynamic and postural Baropodometric step analysis was performed on ten healthy, elderly subjects and ten elderly subjects with arthritis of the knee. The latter group was evaluated both pre- and post prosthetic knee surgery. The data revealed that the ten healthy subjects with arthritis who, prior to surgery presented unequal weight distribution on the diseased side which was slowly redistributed after surgery. PMID- 11892422 TI - [Indications for definitive cardiac stimulation]. AB - Within less than a half-century, after the early rising of cardiac pacing, we witness a dramatical in crease of its indications. After the initial aim, which was to prevent transient ischaemic events, and sudden death due to bradycardia, some more physiological objectives have--progressively appeared, such as improvement of patient's quality of life, and optimization of the cardiac performance to fulfill the metabolic needs. The indications of cardiac pacing are nowadays extended to the fields of haemodynamics and rythmology. Numerous studies are advocating the interest of the cardiac pacing in pathologies such as obstructive and dilated cardiomyopathies specially for the improvement of the NYHA functional status, life comfort and effort sustain. On another hand, newly discovered antiarrhythmic virtues of atrial pacing are of a great interest for a certain type of atrial fibrillations such as vagal induced fibrillations, atrial diseases and atypical flutters. For conclusion: after becoming mandatory for bradycardias, cardiac pacing is conquering new indications on the fields of arrhythmias and cardiomyopathies. Within a close future, scientific evidences could definitely validate cardiac pacing using on these new fields. PMID- 11892421 TI - The use of subjective rating of exertion in Ergonomics. AB - In Ergonomics, the use of psychophysical methods for subjectively evaluating work tasks and determining acceptable loads has become more common. Daily activities at the work site are studied not only with physiological methods but also with perceptual estimation and production methods. The psychophysical methods are of special interest in field studies of short-term work tasks for which valid physiological measurements are difficult to obtain. The perceived exertion, difficulty and fatigue that a person experiences in a certain work situation is an important sign of a real or objective load. Measurement of the physical load with physiological parameters is not sufficient since it does not take into consideration the particular difficulty of the performance or the capacity of the individual. It is often difficult from technical and biomechanical analyses to understand the seriousness of a difficulty that a person experiences. Physiological determinations give important information, but they may be insufficient due to the technical problems in obtaining relevant but simple measurements for short-term activities or activities involving special movement patterns. Perceptual estimations using Borg's scales give important information because the severity of a task's difficulty depends on the individual doing the work. Observation is the most simple and used means to assess job demands. Other evaluations integrating observation are the followings: indirect estimation of energy expenditure based on prediction equations or direct measurement of oxygen consumption; measurements of forces, angles and biomechanical parameters; measurements of physiological and neurophysiological parameters during tasks. It is recommended that determinations of performances of occupational activities assess rating of perceived exertion and integrate these measurements of intensity levels with those of activity's type, duration and frequency. A better estimate of the degree of physical activity of individuals thus can be obtained. PMID- 11892423 TI - [Brugada syndrome]. AB - The Brugada syndrome is a clinical-electrocardiographic diagnosis characterised by syncopal or sudden death episodes in patients with a structurally normal heart with a characteristic electrocardiographic pattern consisting of ST segment elevation in the precordial leads V1 to V3 and a morphology of the QRS complex resembling a right bundle branch block. In many patients with the Brugada syndrome, the electrocardiographic manifestations transiently normalize; leading to underdiagnosis of the syndrome. The administration of sodium channel blockers such as ajmaline, flecainide or procainamide accentuate the ST segment elevation and can be used to unmask concealed and intermittent forms of the disease. The incidence of sudden death in this syndrome is very high and can only be prevented by implanting a cardioverter-defibrillator. Because of high incidence of familial occurrence, the extension of the testing to family members is important. PMID- 11892424 TI - [Diastolic heart failure--echocardiographic aspects]. AB - Diastolic function of the heart is a complex sequence of multiple interrelated events, and it has been difficult to understand, diagnose and treat the various abnormalities of diastolic filling that occur in patients with heart disease. Doppler is used to examine the different diastolic filling patterns of left ventricle. This concept can be applied clinically to estimate left ventricular filling pressures and to predict prognosis in selected groups of patients. This review presents a simplified approach to understanding the process of diastolic filling of the left ventricle and interpreting the Doppler flow velocity curves as they relate to this process. PMID- 11892425 TI - [Percutaneous mitral commissurotomy. 5-year results]. AB - Long-term results of percutaneous mitral commissurotomy were evaluated in 410 patients with mean age of 31 years (18 to 68 years). 48% of patients had mean thickened leaflets, 35% had calcified valves and 17% had flexible leaflets and subvalvular apparatus. Procedure was performed with a double balloon in 57% and with Inoue's balloon in 43% patients. A good immediate results was obtained in 77% of patients. A good result was defined as a mitral valve area > or = 1.5 cm2 without mitral regurgitation. Clinical follow-up concern 378 patients. The actuarial 5 years rate were 84% in our serie, without surgery or new percutaneous mitral commissurotomy and good functional results (NYHA class I or II) were 71%. Valvular anatomy, immediate results (mitral valve area), history of mitral commissurotomy, old patients, atrial fibrillation can influence strongly the results. PMID- 11892426 TI - [Percutaneous mitral commissurotomy: how can it be made simpler and more efficacious?]. AB - The authors report the results of a simplified method of percutaneous mitral dilatation from 936 procedures. This method introduced in our service since 1997 consists in progressively increasing diameters inflation of Inoue balloon, using echographic control and without any arterial procedure. Our population consists on 683 women (73%) and 253 men with a mean age 34 +/- 15 years (extremely 9 and 80 years) with subgroups composed of 26 pregnant women, 32 children less than 17 years, 59 restenosis after closed heart mitral commissurotomy, 14 restenosis after open heart mitral commissurotomy, 74 restenosis after percutaneous mitral commissurotomy, 5 tricuspid and mitral dilatation, 2 coronary and mitral dilatation (with stenting), 2 patients with a history of surgical mitral valvuloplasty for pure mitral regurgitation. The mitral area passed from 1 +/- 0.2 cm2 to 2.1 +/- 0.1 cm2. A surgical mitral regurgitation was observed in 3 cases. The time of fluoroscopy was 6.4 +/- 3.3 min. The hole time procedure was 19 +/- 9 min. The delay of hospitalisation was one day in 97% of patients. The decreased cost was about 20%. The patient comfort and the large diffusion of this method predicts a good future of this simplified technique. PMID- 11892427 TI - [Determining factors in mitral valve reconstructive surgery. Report of 150 cases]. AB - 150 patients had mitral valve repair for mitral valve incompetence. There were 57 males and 93 females with a mean age of 22 years. 60% of the patients were in Class II NYHA and 40% in Class III and IV. Type I was present in 18 patients, type II in 98 and type III in 34 cases. Mitral repair included correction of valve prolapse, valvular enlargement with pericardial patch and annuloplasty with semi-rigid ring in 128 cases and PTFE patch along the posterior leaflet in 12 cases. The perioperative mortality rate was 2.6%. All patients had early post operative echocardiography. According to this, mitral regurgitation was absent or tiny in 135 patients, grade II in 10 cases and grade III in 2 cases. It was moderate or important in twelve patients. In the late post-operative period. All the others patients were reoperated upon for mitral dysfunction in a mean time of 37 months. The reason for reoperation was in the majority of the cases the recurrence of mitral regurgitation related to increase of valvular and sub valvular disease. The late mortality rate is 7%. Out of 126 reviewed survivors on the long run, 71 patients are asymptomatic in class I, 53 patients in class II and 2 patients in class III NYHA. The estimated mitral regurgitation by echography in those patients is absent or trivial in 96 cases. grade II in 29 cases and grade III in one case. Mitral valvuloplasty is the preferred procedure in mitral insufficiency surgical management. It is associated to a low early mortality and morbidity rate. Despite the need for reoperation in about 10% of the cases in the long follow-up, mitral repair offers a good quality of life without anticoagulant treatment. PMID- 11892428 TI - [Unstable angina: study of in-hospital and short-term prognosis]. AB - The aim of our study was to identify predictor factors of coronary ischemic events to stratify the risk. 367 patients, mean age 59 years (22-90), 288 men (88%), Coronary risk factors: Smoking (62%), diabetes (38%), hypertension (37%), hypercholesterolemia (18%). BRAUNWALD class III was predominant (60%). Electrocardiographic changes were present in 113 patients. Coronary angiography identified: 148 single-vessel disease, 92 double-vessel and 68 triple-vessel. In the hospital phase, 296 patients (80.5%) were stabilised. 65 had recurrent ischemia (17.5%), 6 myocardial infarction (1.5%) and 6 deaths (1.5%). After multivariate logistic regression the predictors factors of ischemic events were. Age > or = 65 years (p = 0.03), coronary artery bypass grafting (p = 0.05), left ventricular failure (p = 0.024), modified baseline electrocardiogram (p = 0.04), ST-segment depression (p = 0.05), without aspirin (p = 0.043) and heparin (p = 0.047). At 6 months, 181 patients were asymptomatic (59.1%), 101 had recurrent ischemia (33%), 14 myocardial infarction (4.6%) and 10 deaths (3.3%). After multivariate analysis, the predictor factors were: Age > or = 65 years (p = 0.026), previous unstable angina (p = 0.023), left main stenosis (p = 0.008) and without aspirin (p = 0.003). CONCLUSION: Our study identified a subgroup of high risk patients who would benefit most from either low-molecular-weight heparins and Glycoprotein IIb/IIIa receptor blockers with an early revascularisation strategy. PMID- 11892429 TI - [Predictive factors of normal coronary angiography]. AB - The aim of this study is to compare two groups of patients Group A consisted of 120 patients (70 men and 50 women) hospitalised for anginal symptoms, with either clinical or electrical positive exercise test and/or ischemic events on a 24 H electrocardiography and having angiographically normal coronaries Group B consisted of 120 patients (102 men and 18 women) hospitalised for an acute coronary syndrome with pathological coronaries. The analysis of the 2 groups showed that in the group A the average age was lesser (56 years vs 60 years), women's percentage was higher (41% vs 15%) and cardiovascular risk factors were less frequent. Data from non invasive tests was significantly different in the 2 groups: the exercise test showed both clinical and electrical ischemic events in 35% of the patients in group A versus 75% in group B (p < 0.01) and the 24 h electocardiography showed ST depression in 9% of patients in group A versus 25% in group B (p < 0.01%). The coronary angiography is an invasive and an expensive procedure. The results of our study allow us to modulate its indications, especially in young women patients, with few or no cardivascular risk factors and with only electrical positive exercise test. PMID- 11892430 TI - [Atrial fibrillation]. AB - The aim of transesophageal echocardiography (TOE) in atrial fibrillation milieu is to search a cardiac chamber thrombus. In order to establish the indications of TOE and to raise new issues for the echographic prognosis of maintain of sinus rhythm, 40 patients with atrial fibrillation who underwent cardioversion were prospectively screened. All patients had transthoracic and transesophageal echocardiography before procedure to control the absence or disappearing of atrial thrombus (N = 7). All 40 patients underwent a successful cardioversion. Follow-up was done after one, three and six months for both success group (N = 28) and refibrillation group (N = 12). Our study provides evidence that the only predictive echographic factor of maintain of a good result (sinus rhythm) after cardioversion was a systolic velocity peak > 0.25 metir/sec. PMID- 11892431 TI - [Immediate postoperative arrhythmias follwing pneumonectomy for lung cancer]. AB - Immediate postoperative arrhythmias after pneumonectomy for non small cell lung cancer is a serious complication. Frequency is estimated 10 to 28% of all patients. The goal of this study is to evaluate the incidence of this complication in our experience, preoperative risk factors, therapeutic implications and short outcome. 132 consecutive patients underwent pneumonectomy for lung cancer. We retrospectively studied this series of which 29 patients developed arrhythmias postoperatively. Mean age was 58 years (48 to 79), 16 patients were older than 65 years. Seven patients had medical history of either myocardial infarction or hypertension. Arrhythmias appeared post-operatively on days 1 and 2 six times, days 3 ans 4 ten times, days 5 to 6 six times and days 7 to 10 twice. The trouble consisted in atrial fibrillation in 18 patients, atrial associated with ventricular premature beats in 11 patients. Antiarrhythmic medication (amiodarone) was started as soon as the trouble was confirmed by EKG in all cases. Normalization was obtained in 27 patients (95%). One patient remained dysrrhythmic in spite of treatment. One death occurred on day 4 postoperatively. The mean hospitalisation stay was 10 days. CONCLUSION: Cardiac arrhythmia in the immediate postoperative course is not rare. Early diagnosis in patients at risk followed by adequat treatment is necessary to avoid haemodynamic storm in these quite fragile patients. Amiodarone is the treatment of choice. PMID- 11892432 TI - [Dumesnil's method: is it viable?]. AB - BACKGROUND: Left ventricular (LV) ejection fraction (EF) is an indicator of left ventricular systolic function and is a potent predictor of cardiovascular mortality. LVEF is assessed by a variety of methods, however echocardiography is the most used in clinical practice. Simpson biplane multiple disc method (BMDM) is recommended by the American Society of Echocardiography; Dumesnil's method based on doppler echocardiography seems to be simpler and theoritically less influenced by distortion of LV geometry. OBJECTIVE AND METHODS: To assess the accuracy and reproducibility of Dumesnil's method a group of 100 patients proposed for coronarography with left ventricular angiography, prospectively underwent LVEF measurements by both BMDM and Dumesnil's method. RESULTS: Compared with LV angiography, the correlation coefficient for the Dumesnil's method was r = 0.85 and it was r = 0.9 for BMDM. Correlation in patients with LV regional asynergy were respectively r = 0.69 and r = 0.85. Intraobserver and interobserver variabilities were less then 7% for both echocardiographic methods. CONCLUSION: Although Dumesnil's method is less accurate than BMDM, it is simpler, more rapid with a satisfactory reliability and reproducibility. PMID- 11892433 TI - [Percutaneous closure of ostium secundum atrial septal defect using an Amplazter device. Report of the first 5 Tunisian cases]. AB - We report our initial experience with percutaneous closure of ostium secundum atrial septal defect using Amplatzer device. Between september 2000 and july 2001, five devices were implanted in 5 patients, 4 female and 1 male. Age ranged from 18 to 66 years. A large atrial septal defect with significant shunting was diagnosed by trans thoracic echocardiography. Procedures were performed under general anesthesia with trans esophageal echocardiography guidance. Stretched diameter of atrial septal defects was determined by balloon sizing, ranged from 21 to 32 mm. Amplatzer devices of 20 to 32 mm were respectively deployed. No complications occurred during the procedure. Total occlusion of interatrial communication, was observed in one patient, immediately after the procedure, and in 3 patients 24 hours later. First 3 patients were controlled at months follow up were free of complications. Closure of atrial septal defect with Amplatzer device appears feasible and safe. However, long term results in comparison to surgery remain to be determined before definite conclusion regarding its use can be made. PMID- 11892434 TI - [Cardiac hydatid cyst (report of 8 cases)]. AB - Hydatic cardiac cyst (HCC) is a rare anomaly, characterized by wide spectrum of clinical presentations. Its diagnosis took large benefit from echocardiography progress. We aimed in this study to analyse epidemiologic aspect of HCC, as well as its diagnostic data, with special emphasize on echocardiography. Observations of 8 patients, admitted between 1991 and 2000 has been reviewed. Age average was 30 years (13 to 59). Main symptoms consisted of chest pain (62.5%) and dyspnea (35%). Chest X-Rays showed cardiac silhouette anomaly in 4 cases. Electrocardiogram showed sub-epicardical ischemia in 5 patients. HCC diagnosis was established by transthoracic echocardiography in all cases. It was located inside a cardiac chambers (n = 5), the interventricular septum (n = 2), and the pericardium (n = 1). HCC relation with adjacent cardiac structures was well defined by transoesophageal echocardiography performed in 7 cases. All patients underwent surgery with good immediate results. All patients were events free, with a mean follow-up of 25 months. PMID- 11892435 TI - [The role of Doppler echocardiography in chronic constrictive pericarditis]. AB - Constrictive pericarditis (CCP) is a rare but serious disease. It still poses diagnostic difficulties. The purpose of our work is to study the contribution of the echocardiographic Doppler in the diagnosis of the CCP. The authors report six cases of CCP proven after surgery. Study by ultrasound Doppler of intracardiac blood flow and their respiratory variations showed the existence of abnormalities. The decrease of 25% of the mitral E wave in inspiration compared to the value observed in expiration, the increase of 100% of the ebb in sus hepatic vein in expiration and the modifications of the flux in pulmonary insufficiency are the most reliable signs for the diagnosis of the CCP. This method seems so interesting for the diagnosis and to estimate the degree of constriction of the CCP. PMID- 11892437 TI - [Indications for radiotherapy of benign lesions: yesterday, today and tomorrow]. AB - A better knowledge of the radiobiological effects and the control of the techniques of dosimetry led to a renewed interest for the radiotherapy of the benign lesions. The doses used for these indications are weaker than those recommended for treatment of cancer and the radiobiological mechanisms implied are different. The aim of this review of the literature is to specify the radiobiological mechanisms, the risks and the place of ionizing radiations during the processing of the benign lesions. Although the risk of radiation induced neoplasms remains a limiting factor of the indications, those are very varied. Some indications are well accepted such as keloid, cerebral arteriovenous malformations, graves' ophtalmopathy, prevention of postoperative heterotopic bone formations; and some others remain still controversial such as the prevention of the post angioplasty restenosis and age-related macular degeneration. PMID- 11892436 TI - [Bacillus anthracis: causative agent of anthrax]. AB - Anthrax, an acute infectious disease of historical importance, is once again regaining interest with its use as a biological weapon. It is caused by B. anthracis, a Gram positive spore forming rod usually surrounded by a capsule and producing toxin. It occurs most frequently as an epizootic or enzootic disease of herbivores that acquire spores form direct contact with contaminated soil. Spores can survive for many years in soil. Animal vaccination programs have reduced drastically the disease in developed countries. In humans, the disease is acquired following contact with anthrax infected animals or their products. 3 types of anthrax infection can occur: cutaneous, inhalational and gastro intestinal. Cutaneous anthrax is the most common observed form. When germination occurs, replicating bacteria release toxin leading to hemorrhage, edema, necrosis and death. Full virulence of B. anthracis requires the presence of both antiphagocytic capsule and 3 toxin components (protective antigen, lethal factor and edema factor). Most naturally occurring anthrax strains are sensitive to penicillin but resistant to third generation cephalosporins. Post exposure prophylaxis is indicated to prevent inhalational anthrax. PMID- 11892438 TI - [Systemic manifestations of chronic hepatitis C]. AB - It's well known that hepatitis C virus (HCV) related chronic liver disease may be associated with various extra hepatic disorders. These manifestations can revealed the hepatic disease. We review the available data on the conditions and asses their clinical implications: vascular, cutaneous, articular, neurological or renal disorders. There is no correlation between these extra hepatic manifestations and the severity of liver disease. Several recent studies have established a strong link between HCV infection and essential mixed cryoglobulinemia but some other extra hepatic associations are just fortuitous. Others datas are necessary to better analyze these extra hepatic disorders and to offer the beneficial treatment of patients with chronic hepatitis C. PMID- 11892439 TI - [Severe pulmonary hypertension in ostium secundum atrial septal defect]. AB - Between 1969 and 1999, 149 patients underwent cardiac catheterization for ostium secundum atrial septal defect, in hemodynamics department. Age average was 21.5 +/- 16 years (2-67). Systolic pulmonary arterial pressure average was 35 +/- 15.6 mm Hg (15-140). The atrial septal defect was closed in 108 patients (by surgery in 107 cases, and by interventional catheterization in one case). Mean follow-up was 8 +/- 7 years (1-20). Overall mortality rate was 2.7%. Severe pulmonary hypertension (systolic pressure > or = to 50 mm Hg) was seen in 13.4% of cases. It was present at all ages, but more frequent above 20 years. Obstructive pulmonary vascular disease was noticed in 2% of patients. Clinical and haemodynamic data analysis showed that, severe pulmonary hypertension is a frequent, and unpredictible complication of secundum atrial septal defect; it might occur at any age and worsen considerably the cardiac disease prognosis. Because of atrial septal defect hemodynamic profile, and current rise of percutaneous closure technics, invasive investigation should be more and more routinely performed, in order to define the best therapeutic option. Early and systematic closure of secundum atrial septal defects, should prevent, or at least minimize pulmonary vascular lesions. PMID- 11892440 TI - [Epidemiology and development of macular edema in the diabetic]. AB - Macular edema is the first cause of blindness in diabetics. Macular edema is defined by macular thickening or deposits of hard exudates. On 1000 diabetics examined over 2 years, 60 patients had a macular edema of which we retained 38 cases(54 eyes). All the patients had an ophthalmologic examination with a retinal angiography. Laser photocoagulation with green Argon laser was instituted in 50 eyes. 63% had background rethinopathy. Total or partial regression of the edema happened in 84.4%. Laser photocoagulation decrease by the half vision loss risk. Interest of early detection and treatment to decrease blindness incidence of macular edema in diabetics. PMID- 11892441 TI - [Epidemiology of hepatitis C virus infection in the Menzel-Bourguiba region]. AB - This study include 151 serum samples HCV + which has been diagnosed during 3 years. The number of patients HCV + (110) is increasing every year, but the rate of blood donors HCV + remain stable (1.18%). An epidemiological study was made about 81 patients HCV + from gastroenterologic unit. It appears that these patients are old (60.4 years on average), often women (ratio H/F = 0.68), and treated for chronic diseases. The source of infection usually known (blood transfusion, hemodialysis, surgical operation,...) are not predominant (33.2%), and we think about nosocomial transmission. The serotype 1 is predominant (60.7%), but he is associated with poor therapeutic efficiency. There is not vaccine, and obviously preventive measures are essentials to limit the progression of HCV infection. PMID- 11892442 TI - [Vaccination against viral hepatitis B for public health personnel: the case of the National Fund of Social Security]. AB - Viral hepatitis B is endemic in Tunisia at an intermediate level. Health professional are a group particularly exposed to the infection. Immunization is the most efficient way to prevent this disease. In this study, we had try to estimate the cost of three strategies of immunization: immunization after a complete serology, immunization after a sequencial serology and immunization without previous serology. The study was conducted at the clinic of the National Fund of Social Security in El Khadra. All the personal of the clinic was invited to participate to a program of immunization after complete serology. Participation rate was at 93.8%. 33.3% of the personal was immunized (24% by a previous contact with the virus and 9.9% by a previous immunization). The costs of the three strategies were as below: immunization after complete serology: 53.4 tunisian dinars by person. immunization after sequantial serology: 33.06 tunisian dinars by person. immunization without serology: 16.2 tunisian dinars by person. Immunization without previous serology has the lower cost, and doesn't expose at any sid effect. However, a post immunization serology vaccinal should be desirable. PMID- 11892443 TI - [Hairy cell leukemia: report of 8 cases]. AB - Hairy cell leukemia haemopathy is a rare lymphoid haemopathy type B. 8 cases are reported and diagnosed at Hopital Aziza Othmana over a period of 20 years between 1979 and 1999, 7 men and one women. The mean age of the patients is 51 years, with externe ages from 42 to 81 years. 4 patients consulted for an infections and, or anaemia syndrome. The disease was revealed due to the presence of an isolated splenomegaly in other cases. At the clinical examination, the spleen is hypertrophied in 7 patients out of 8. Pancytopenia is observed in 50% of the patients. Only one patient has presented a moderated hyperleukocytosis at 11,000/mm3 related to the presence of moving on tricholeukocytes. The myelogramme is pocr. It allowed to mention the diagnosis in 6 cases out of 8. Bone Marrow biopsy revealed a diffuse infiltration by TCL with a reticulinic fibrosis in all patients. 4 patients out of 8 have been splenectomized. Cytopenies have been corrected in all patients. Only one patient has been treated by alpha Interferon for 3 years with a partial hematological response. A relapse was observed once the Interferon was stopped. With the introduction of new drugs such purine analogues. The HCL treatment has been revolutionarized thanks to the improvement of the rate of complete response (from 10% to 80% of CR). If splenectomy is still observed in HCL for splenomegalic and or severe cytopenia, our findings could be improved thanks to new purine analogues. PMID- 11892445 TI - [Heterotopic pregnancy: 3 case reports]. AB - Heterotopic pregnancy is a rare event combining intra and extra uterine pregnancies. We report 3 cases observed in patients aged 32, 32 and 31 years consulting for pelvic pain and metrorragia with amenorrhea of 6 to 7 weeks. Treatment was conservative in 2 cases and radical for the third patient. The frequency of this association has been increased since the development of medical procreation technique and the increased of ectopic pregnancy. The foetomaternal prognosis will be improved by an early diagnosis. PMID- 11892444 TI - [The HELLP syndrome: report of 11 cases]. AB - Being an independent entity or a type of eclampsia, the HELLP syndrome always represent a serious complication of the renovascular disease. This retrospective study realised during five years revealed 11 cases including 3 cases occurring in the post partum. For all this patients, this complication happened during the 3rd term of pregnancy. The mean period of gestation when it occurred was the 34th week of pregnancy. In 36% of cases, the discovery of the HELLP syndrome was fortuitous by routine laboratory tests. The symptoms and signs are dominated by the digestive ones. In case of severe maternal complication or when the foetal lungs reach maturity, the interruption of pregnancy is the rule. In the other cases, expectative attitude with an intense observation may be proposed when the 32nd week of pregnancy isn't reached. PMID- 11892446 TI - [Epiphyseal detachment of the upper end of the tibia]. AB - In this study, we report 3 cases of fractures involving separation of the upper end of the tibia in young adolescent. These are fairly rare injuries that usually treated by plaster immobilisation. Our aim, through this work is to show the uncommon characteristic of this injuries, possibility of local complications and the difficult of choice of treatment: conservative or surgical treatment. PMID- 11892447 TI - [Duplicate principal bile ducts. Two case reports]. AB - The duplicity of the way biliaire main is a rare abnormality. We report two observations revealed by episodes of angiocholite. The degrades cholangiopancreatography endoscopy allowed to wear diagnosis and to practise a therapeutic gesture. PMID- 11892449 TI - [Medical confidentiality from the clay tablet to the optic fiber]. AB - Medical confidentiality is as old as medicine. Its protection has been variabe through ages. Hippocrate's Oath is considered to be the first reference to physician's duty to protect medical confidentiality. Arabic and Muslim medicine strengthened this obligation. Conflicts between community and patient's private interests led to derogations allowing medical record disclosing. Nowadays, protection of medical confidentiality is emphazised in charts and international codes related to patients rights. PMID- 11892448 TI - [Basal cell carcinoma arising in a seborrheic keratosis: a case report]. AB - Seborrheic keratosis are one of the most common benign epidermic tumors in clinical practice. Malignant transformation is exceptional and occurs by the involvement of human papilloma virus. We report a case of seborrheic keratosis of the armpit in a 55 year-old woman whose biopsy revealed the presence of a basal cell carcinoma. PMID- 11892450 TI - Identifying environmental factors related to patient safety. PMID- 11892451 TI - America under attack: ACHE affiliates respond. AB - In the midst of the horror and uncertainty that swept over America on September 11, the healthcare sector helped to keep our nation firmly anchored. Within moments of the terrorist attacks, healthcare organizations in New York, Washington, D.C., and the surrounding areas responded swiftly, calmly, and effectively. Many of these hospitals are led by ACHE affiliates. Following are their accounts of that day, lessons they learned, and plans for the future. PMID- 11892452 TI - Effective top teams: five strategies for success. PMID- 11892453 TI - Adding value. PMID- 11892454 TI - The values of a profession. PMID- 11892455 TI - Interview with ACHE's incoming chairman. PMID- 11892456 TI - Partnership for success. PMID- 11892457 TI - Beyond hospital walls. PMID- 11892458 TI - 2001 Federal health policy activities. PMID- 11892459 TI - Establishing competencies for healthcare managers. PMID- 11892461 TI - Attracting and keeping employees. PMID- 11892460 TI - Principle-based governance. PMID- 11892462 TI - Preserving and restoring patient trust. PMID- 11892463 TI - Data 'cubes' help health plan quickly spot variation. PMID- 11892465 TI - Martin's Point gathers feedback from patients through intranet. AB - A Maine not-for-profit health system views comments from its 54,000 patients as an important source of information about how health care quality can be improved. Submitting comments is as easy as clicking a computer mouse. PMID- 11892464 TI - Homegrown automated anesthesia record puts Virginia hospital on cutting edge. PMID- 11892466 TI - Patient registry provides benchmarks for treatment of myocardial infarction. AB - Mercy General Hospital in Sacramento continually tracks its treatment of MI patients against national benchmarks. The result is care that consistently meets or exceeds the nation's best. PMID- 11892467 TI - [Tissue Doppler echocardiography: a new technique to assess diastolic function]. AB - Diastolic dysfunction and elevated filling pressure have important role in heart failure. Traditional Doppler echocardiography (DE) however is of limited value in the measurement of these variables. The objective of this study was the evaluation of a new technique, the pulsed tissue Doppler echocardiography of the lateral mitral anulus (PTDI) in diastolic function. 96 consecutive patients were included into the study who were in sinus rhythm, mitral valve disease was excluded and ejection fraction was either > or = 50% (50 patients) or < or = 40% (46 patients). DE technique was used to measure mitral E, A velocity, deceleration time (DT). Myocardial early diastolic (Ea), late diastolic (Aa) velocities were measured at the lateral mitral anulus by PTDI, and E/A, Ea/Aa, E/Ea ratios were calculated. Based on accepted DE criteria of diastolic dysfunction there were 19 normal subjects, 18 patients had abnormal relaxation, 8 had pseudonormalization pattern and 12 had restrictive dysfunction, the rest of patients did not fulfill these criteria. RESULTS: PDTI indicated an association of diastolic dysfunction to systolic dysfunction, which could not be shown by DE. Myocardial Ea velocity was age-dependent only in patients with good systolic function, and it was less than 15 cm/s in all types of diastolic dysfunction. E/Ea ratio over 8 indicated elevated filling pressure, but it was related to E/A ratio only in cases of good systolic function. Importantly one third of cases could be classified into diastolic dysfunction patterns only using tissue Doppler. CONCLUSIONS: Myocardial diastolic velocities can be easily measured by pulsed tissue Doppler technique at the lateral mitral anulus. Decreased early diastolic tissue velocity indicates diastolic dysfunction independently of its type, and it is generally associated to systolic dysfunction, independently of age. Pseudonormalization is defined as the combination of normal mitral inflow and decreased tissue diastolic velocity. PMID- 11892468 TI - [Amyopathic dermatomyositis]. AB - Dermatomyositis is a systemic autoimmune disease which belongs to the group of idiopathic inflammatory myopathies. The disease is rare with an incidence of 0.1 1/100,000 and a prevalence of 1-6/100,000. Women are affected twice as often as men. In some patients the disease presents with dermatologic changes weeks to months before the myopathy arises. It was observed that in some patients the myositis develops much later and sometimes not at all. Therefore, the term of dermatomyositis sine myositis used in earlier literature has been changed to amyopathic dermatomyositis. The most important question is whether the patient needs systemic therapy with its possible side effects yet possibly preventing the appearance of myositis or only local therapy for the skin manifestations is necessary. The goal of this article is to summarize the latest findings in amyopathic dermatomyositis. PMID- 11892469 TI - [Treatment of acute leukemia in older patients]. AB - The median age of patients with acute leukemia is more than 60 years, and the incidence of the disease increases with age. There are several unfavorable biologic and clinical factors in older patients with acute leukemia, the remission rate and their survival are much worse than in younger adults. The poor performance score and any other concomitant disease and the poor reserve capacity of their bone marrow represent significant difficulties to deliver the proper remission induction and postremission therapy. The poor biologic factors like acute leukemia following myelodysplasia, unfavorable cytogenetic abnormalities and multidrug resistance among elderly patients are more common comparing to younger patients. The authors summarize the international data and attempt to define the proper therapy namely who are candidates for induction and intensive postremission therapy, what is the role of palliative cytostatic treatment or supportive care for these patients. PMID- 11892470 TI - [Sapporo-like viruses in sporadic gastroenteritis of unknown origin]. AB - Sapporo-like viruses (SLVs) are members of the family Caliciviridae. The etiologic role of these viruses is evident but little is known about the incidence of acute gastroenteritis caused by them. Sapporo-like viruses have not been detected in Hungary before. METHODS: Between October and December 2000, 72 sporadic diarrhoeal stool samples from Baranya County, Hungary, from infants and young children (under 12 years) with acute gastroenteritis were collected. Common enteric bacterial pathogens, adeno- and rotaviruses were not found in these stool samples. Reverse transcription-polymerase chain reaction with specific primer pairs for human caliciviruses were used to detect Sapporo-like viruses. RESULTS: The amplicons of expected size were cloned and subsequently sequenced. Seven (9.7%) of 72 stool samples were found to be positive for Sapporo-like viruses. Comparative sequence analysis confirmed that all strains belonged to the London/92/UK cluster. CONCLUSION: This is the first molecular detection and molecular sequence analysis of Sapporo-like viruses in Hungary. PMID- 11892472 TI - [The journal Gyogyaszat - 100 years ago]. PMID- 11892471 TI - [The Hungarian Secretary of the Department of Health in Liberia]. PMID- 11892473 TI - Stop press: things are getting better. PMID- 11892474 TI - Hands up for hands-on. PMID- 11892475 TI - Quality care? Enquire within. PMID- 11892476 TI - After the fall, the fallout. PMID- 11892477 TI - Is the nurse consultant post working as intended? PMID- 11892478 TI - No time to draw breath. PMID- 11892479 TI - 'A real opportunity to make a difference'. PMID- 11892480 TI - An infectious enthusiasm. PMID- 11892481 TI - The year of living conspicuously. PMID- 11892482 TI - Students' views on clinical placements. PMID- 11892483 TI - Supporting newly qualified nurses in operating theatres. PMID- 11892484 TI - Palliative care in acute hospitals. PMID- 11892485 TI - Client-centred goal planning. PMID- 11892486 TI - A service to help families deal with eating disorders. PMID- 11892487 TI - HIV and AIDS. 2. Modes of transmission, testing for HIV antibodies, and occupational exposure to HIV. PMID- 11892488 TI - We can afford new antimicrobials for all. PMID- 11892489 TI - Evidence for infectious agents in cardiovascular disease and atherosclerosis. AB - During the past decade, several novel risk factors for atherosclerosis, including inflammation and infections, have been reported. Seroepidemiological studies suggest an association between several microbes and coronary heart disease. Microbes or their structural components are found in atherosclerotic plaques, but the only intact microbes commonly present are herpes viruses and Chlamydia pneumoniae. These agents are able to initiate and accelerate atherosclerosis in animal models. If they cause persistent infection in the vessel wall, they can directly promote a proinflammatory, procoagulant, and proatherogenic environment. Microbes could also have a remote effect--e.g., bacterial heat shock proteins with high sequence homology with human counterpart could, in the presence of a chronic infection, induce autoimmunity against vascular cells, and lead to an atherosclerotic process. Several intervention trials with antibiotics are underway, and will hopefully shed new light on the role of bacteria in atherosclerosis. The causal relationship can be proved by use of vaccination to prevent infections. PMID- 11892490 TI - Novel methods for the detection of microbial antibodies in oral fluid. AB - Compared with blood, oral fluid has several advantages as a sample for antibody detection. It is simple, safe, painless, and cheap to collect. The only drawback is that while the antibody profiles indicate those in blood, they are at lower concentrations. Antibody capture assays are the method of choice for the detection of microbial antibodies in oral fluid, but their relative lack of sensitivity when based on conventional immunoassay techniques has mostly limited their use to epidemiological applications. Immuno-PCR and time-resolved fluorescence offer more sensitive detection systems that could be applied to oral fluid specimens. We review antibody detection in oral fluid and discuss immuno PCR and time-resolved fluorescence as candidate systems. Both have the potential to broaden the applications of oral fluid testing to clinical diagnostics. PMID- 11892491 TI - US not ready to destroy smallpox stocks. PMID- 11892492 TI - Anthrax concerns persist in the USA. PMID- 11892493 TI - Estimates of world-wide distribution of child deaths from acute respiratory infections. AB - Acute respiratory infections (ARI) are among the leading causes of childhood mortality. Estimates of the number of children worldwide who die from ARI are needed in setting priorities for health care. To establish a relation between deaths due to ARI and all-cause deaths in children under 5 years we show that the proportion of deaths directly attributable to ARI declines from 23% to 18% and then 15% (95% confidence limits range from +/- 2% to +/- 3%) as under-5 mortality declines from 50 to 20 and then to 10/1000 per year. Much of the variability in estimates of ARI in children is shown to be inherent in the use of verbal autopsies. This analysis suggests that throughout the world 1.9 million (95% CI 1.6-2.2 million) children died from ARI in 2000, 70% of them in Africa and southeast Asia. PMID- 11892494 TI - Dengue: an update. AB - This review is an update of dengue and dengue haemorrhagic fever (DHF) based on international and Cuban experience. We describe the virus characteristics and risk factors for dengue and DHF, and compare incidence and the case fatality rates in endemic regions (southeast Asia, western Pacific, and the Americas). The clinical picture and the pathogenesis of the severe disease are explained. We also discuss the viral, individual, and environmental factors that determine severe disease. Much more research is necessary to clarify these mechanisms. Also reviewed are methods for viral isolation and the serological, immunohistochemical, and molecular methods applied in the diagnosis of the disease. We describe the status of vaccine development and emphasise that the only alternative that we have today to control the disease is through control of its vector Aedes aegypti. PMID- 11892495 TI - Immunology of hepatitis B infection. AB - The immune response initiated by the T-cell response to viral antigens is thought to be fundamental for viral clearance and disease pathogenesis in hepatitis B virus (HBV) infection. The T-cell response during acute self-limited hepatitis B in people is characterised by a vigorous, polyclonal, and multispecific cytotoxic and helper-T-cell response. By contrast, the immune response in chronic carriers, not able to eliminate the virus, is weak or undetectable. Thus a dominant cause of viral persistence could be the existence of a weak antiviral immune response. Methodological progress in animal models allows more precise investigation of the mechanisms by which the immune system resolves viral infection or develops chronic infection. Although clearance of most virus infections is widely thought to indicate the killing of infected cells by virus-specific T cells, data suggest that non-cytolytic intracellular viral inactivation by cytokines released by virus-inactivated lymphomononuclear cells could have an important role in the clearance of this virus without killing the infected cell. Additional factors that could contribute to viral persistence, which have been partly proven in animal models, are viral inhibition of antigen processing or presentation, modulation of the response to cytotoxic mediators, immunological tolerance to viral antigens, viral mutations, and infection of immunologically privileged sites. In view of the central role of cellular immunity in disease pathogenesis, strategies have been proposed to manipulate this cellular immune response in favour of protection from disease. PMID- 11892496 TI - Overcoming challenges to the implementation of antiretroviral therapy in Kenya. AB - Dramatic price reductions make antiretroviral drugs increasingly affordable in Africa. However, poor infrastructure, conflicting interests, and lack of commitment to a common strategy could keep them beyond the reach of most infected people. Recent experiences in Kenya, where the political will exists, show the complexity of turning this opportunity into safe and effective practice. PMID- 11892497 TI - Robert G Ridley--realising the potential of public-private partnerships. Interview by Pam Das. PMID- 11892499 TI - Infectious disease training: challenges and opportunities. PMID- 11892500 TI - Anesthesia-related cardiac arrest in children. An update. AB - The improvement in mortality rates for anesthetized children over the past 50 years reflects the many improvements that have been made in pediatric perioperative care. The modern pediatric anesthesiologist is better trained than the predecessors of half a century ago, and has a vastly improved arsenal of monitoring devices and anesthetic agents from which to choose. The modern pediatric perioperative environment is better equipped to meet the unique needs of children. Techniques practiced by surgeons, nurses, radiologists, and pharmacologists help create a far more sophisticated infrastructure than existed 50 years ago. Given these changes, it is not surprising that outcomes for patients have improved. PMID- 11892501 TI - New concepts in acute and extended postoperative pain management in children. AB - Increased knowledge of the pathophysiology of pain in children and an improved understanding of the pharmacology and pharmacodynamics of multiple agents have provided the clinician with a wide variety of tools to treat postoperative pain in children. The interest in a multimodal approach is kindled by the realization that the combination of a number of therapies can enhance analgesia with fewer untoward side effects. The expertise of other health care professionals should be tapped to open new avenues of treatment. Many therapies still require critical evidence-based evaluations to assess how well they work in larger patient populations. Dedication to research, compassionate patient care, and a willingness to teach the next generation of clinicians will bring us closer to the goal of safe and pain-free surgery. PMID- 11892502 TI - Neuroanesthesia. Innovative techniques and monitoring. AB - Advances in neuromonitoring have provided insights into neurologic function during anesthesia. Despite the limitations and necessary caution when using intraoperative monitors to interpret neural function, these technologies have been definite steps in the right direction for assessing neural integrity and level of consciousness during anesthesia. The techniques discussed minimize the adverse sequelae of a variety of neurosurgical and orthopedic procedures, reducing the morbidity rates/risks in the perioperative period. Furthermore, it is likely that such monitoring will become a standard of care, similar to other monitoring standards such as pulse oximetry and capnography. Accurate and reliable monitoring is essential, and on-going large prospective studies comparing the processed EEG or evoked potential with definable end points in both adult and pediatric populations will be necessary. The use of monitoring, such as the BIS, may improve cost efficiency by reducing the total amount of drug used to maintain anesthesia, as well as enhance recovery. A danger in this process, however, is the potential for public opinion, outside regulatory bodies, or medico-legal implications to drive change and enforce standards of care before appropriate data are available. PMID- 11892503 TI - Pediatric thoracic anesthesia. AB - The anesthesiologist caring for infants and children undergoing thoracic surgery faces many challenges. An understanding of the primary underlying lesion as well as associated anomalies that may impact perioperative management is paramount. A working knowledge of respiratory physiology and anatomy in infants and children is required for the planning and execution of appropriate intraoperative care. Familiarity with a variety of techniques for SLV suited to the patient's size will allow maximal surgical exposure while minimizing trauma to the lungs and airways. Finally, use of regional anesthetic techniques, including epidural anesthesia and analgesia, facilitates optimal postoperative pain control and pulmonary function. PMID- 11892504 TI - Sedation and analgesia in pediatric patients for procedures outside the operating room. AB - Sedation and analgesia in pediatric patients for procedures outside the operating room are becoming more frequent as health care is being driven to be more cost effective and "efficient." Although anesthesiologists may not be directly involved in sedation or analgesia outside of the operating room, there is a high likelihood that they will be asked by their institutions to be integrally involved in creating and supervising sedation policy given that the American Society of Anesthesiologists and the Joint Commission on Accreditation of Healthcare Organizations consider sedation and analgesia as part of a continuum ranging from minimal sedation to moderate sedation and analgesia, deep sedation and analgesia, and, finally, general anesthesia. Further, anesthesiologists will be asked to define, teach, and credential nonanesthesiology practitioners who perform deep sedation because these practitioners are now required to be qualified to "rescue from general anesthesia." PMID- 11892505 TI - Office-based anesthesia for children. AB - The use of office-based surgery and anesthesia will continue to grow. The anesthesia community has embraced the opportunity to become a driving force of office-based surgery and has organized into rapidly growing groups that promote safe practice in the office setting. The Society for Office-Based Anesthesia was developed to continuously improve patient safety and outcomes in office surgery. This group has an active Web site (www.soba.org) that allows for online discussions and widespread participation in working toward the society's stated goal. This Web site may be used as a reference for physicians in the process of considering the move to office-based anesthesia. The advantages of office-based anesthesia are numerous. The financial incentives are tremendous and the convenience to the patient and surgeon is important. For office anesthesia to be successful in children, patient safety, proof of improved outcomes, and family and surgeon satisfaction must be the goals. Anesthesia providers must continue to take active roles in organizing the office environment to ensure that safety is paramount. As the field grows, additional ways to study and improve the overall care children receive in the office should be sought. In the near future, office practice for surgery and anesthesia for children undergoing minor procedures should be a safe and effective alternative to current practices. PMID- 11892506 TI - Anesthesia for fetal surgery. AB - Fetal surgery is the antenatal treatment of fetal malformations that cannot be adequately corrected after birth. Anesthesia for fetal surgery involves two patients, and issues of maternal safety, avoidance of fetal asphyxia, adequate fetal anesthesia and monitoring, and uterine relaxation are important. Communication with the surgeon to determine the surgical approach and need for uterine relaxation allows the anesthesiologist the ability to vary the anesthetic technique. Lessons learned from fetal surgery may help other neonates with life threatening anomalies and may help understand the complex issues related to preterm labor. PMID- 11892507 TI - Ethical dilemmas for pediatric surgical patients. AB - Anesthesiologists are confronted with interesting and sometimes difficult ethical situations in pediatric surgery. They are forced to deal with everything from "do not resuscitate" issues, heroic last-chance surgical efforts, religious and cultural conflicts, disputes among colleagues, and situations that are, at worst, uncomfortable and, at best, miscarriages of duty. It is incumbent on anesthesiologists to learn how to logically and appropriately handle these issues. The pediatric surgical patient requires special consideration in bioethics. This article discusses the principle of autonomy and its ascension in importance in bioethics. The concepts of informed parental permission, assent, and dissent are presented. The authors provide a framework for ethical problem solving, as well as a discussion of judicial decision-making. In addition, several examples of clinical-ethical situations and the processes used for resolutions are discussed. By using a well-reasoned ethical decision-making process, any situation, from the simple conflict to the most serious resuscitation and withdrawal of care issues, may be appropriately resolved. PMID- 11892508 TI - Psychological preparation of the parent and pediatric surgical patient. AB - Some 3 million children undergo anesthesia and surgery in the United States every year; 40% to 60% of these children develop significant behavioral stress prior to surgery. Multiple interventions have been suggested to treat the preoperative behavioral stress responses in children. There is a trend toward reducing both behavioral and pharmacological preoperative interventions aimed at children, perhaps because though there is a consensus that preoperative interventions can be useful, almost no outcome studies have evaluated the effects of these interventions on measurable, clinically "important" postoperative outcomes. More research is needed in this area. PMID- 11892509 TI - Neuromuscular blocking drugs in pediatric anesthesia. AB - There are differences between pediatric patients and adults in the responses to neuromuscular blockers. The results of new studies of neuromuscular blockers in pediatric patients are generally consistent with earlier studies in the type of age-related differences documented. Surprising new results are the side effects, including anaphylaxis, that have been observed, and studied directly, in pediatric patients. Continued evaluation of these drugs in pediatric patients will provide better information regarding the variability of the effects of neuromuscular blockers in healthy and diseased infants and children. PMID- 11892510 TI - Novel concepts for analgesia in pediatric surgical patients. Cyclo-oxygenase-2 inhibitors, alpha 2-agonists, and opioids. AB - Pain has been an understated concern in infants and children. Failure to recognize pain in the past has resulted in undue suffering by infants and children of all ages, but with the introduction of instruments to measure pain and a widespread appreciation of the severity of this problem in this age group, pain is rapidly coming under control. Novel concepts in both old and new analgesics have created safe and effective pain management strategies for infants and children. This review examines three analgesics and their potential roles in infants and children: cyclo-oxygenase (COX)-2 inhibitors, alpha 2-agonists and opioids. PMID- 11892511 TI - Practical pediatric regional anesthesia. AB - In children, regional anesthetic techniques are safe and effective adjuncts to general anesthesia and for postoperative pain relief. Application of the techniques described in this article will contribute to improved care for pediatric patients undergoing surgical procedures. The judicious choice of local anesthetics, along with the blockades of targeted nerves, decrease the need for supplemental analgesics in the recovery phase. PMID- 11892512 TI - Using the student body: college and university students as research subjects in the United States during the twentieth century. PMID- 11892513 TI - "Live clean, think clean, and don't go to burlesque shows": Charles Atlas as health advisor. PMID- 11892514 TI - Therapy and disease concepts: the history (and future?) of antimony in cancer. PMID- 11892515 TI - Science and sexual identity: an essay review. PMID- 11892516 TI - Electrosurgery policy. PMID- 11892517 TI - Emergency endoscopy and decontamination of endoscopic equipment. PMID- 11892518 TI - Sterilisation and traceability of surgical instruments. PMID- 11892519 TI - Introducing clinical supervision. The pitfalls and problems. AB - In July 1999 I published an article in the BJPN entitled 'Introducing Clinical Supervision into the Perioperative Environment' (Smith 1999). I described what I believed to be the successful implementation of clinical supervision to the trained theatre staff. I discussed the perceived benefits of improved communication, assistance with learning, professional development and reflective practice, provision of emotional support, evidence for PREPP, clinical effectiveness and governance, and improvement practice and policy making. Results from many other authors and professional bodies also proclaim both the need for, and the benefits, of clinical supervision for nurses. PMID- 11892521 TI - Handling conflict. PMID- 11892520 TI - Deep vein thrombosis. Incidence and physiology. AB - Deep vein thrombosis is a serious postoperative complication which can delay recovery and extend hospital stay (Autar 1996). A DVT can be asymptomatic and often precedes a pulmonary embolism (PE), which has been found in 10-25% of hospital deaths (Sandler & Martin 1989). Virchow, a nineteenth century pathologist identified three factors which initiate the formation of a DVT. PMID- 11892522 TI - Partnership in head injury rehabilitation. PMID- 11892523 TI - A clever disguise? Associations unite to call for universal PPO standards, but endorsement could hurt JCAHO. PMID- 11892524 TI - Call to arms. Johnson & Johnson launches new initiative in fight to ease nursing shortage. PMID- 11892525 TI - New nursing group divides ANA. AARN (American Association of Registered Nurses) piggybacks on CNA's success, fights for nurse-staffing ratios. PMID- 11892526 TI - Legal victory for AMA's union group. Labor board rules that New Jersey physicians are employees, not supervisors. PMID- 11892527 TI - Who's minding the store? With the feds scaling back, state attorneys general are left to fill antitrust gap. PMID- 11892529 TI - Turn up the volume. More patients mean healthy growth in earnings for HCA, spinoff Lifepoint Hospitals. PMID- 11892528 TI - Ore. upstart takes on rival. PMID- 11892530 TI - Earnings at a premium. Rate increases fuel gains for HMOs hit by rising costs, faltering economy. PMID- 11892531 TI - The path to affordability. Keeping our quality of care means finding out where costs can be reined in. PMID- 11892532 TI - At your service. After years of feeble attempts to improve customer relations, hospitals are turning to the professionals and spending big bucks to keep their patients happy. PMID- 11892533 TI - 2002 Trustees of the Year. Ohio executive works to keep patients, system healthy. PMID- 11892534 TI - Statement of net worth. As executive salaries keep growing bigger, some healthcare industry experts question whether associations are getting value for money. AB - Healthcare associations are paying their executives a little more every year--and officials say it's worth it for their hard work, clout and connections. In 2000, the chief executives of major healthcare associations earned an average annual base compensation of nearly $400,000. But are the associations really getting value for their money? PMID- 11892535 TI - Scully: everything is on the table. Healthcare industry readies itself for a year of knock-down, drag-out lobbying efforts. PMID- 11892536 TI - Fever of unknown source: outpatient evaluation and management for children 2 months to 36 months of age. AB - This month's clinical practice guideline (CPG) review is on fever of uncertain source (FUS), also known as fever of unknown origin (FUO). A hospital-based committee for the Cincinnati Children's Hospital Medical Center developed this guideline to help practitioners evaluate and manage FUS in a logical manner and with a judicious use of antibiotics. This guideline should have a wide interest to practitioners due to the large number of children that annually present with fever. PMID- 11892538 TI - Barriers to preventive health care for young children. AB - PURPOSE: To review the literature on barriers to availability, access, and utilization of preventive health care for young children three to five years of age and their families and to discuss the role of nurse practitioners (NPs) in future research, education, and practice in this area. DATA SOURCES: A comprehensive literature search was conducted of online material and CINAHL and Medline (CD-ROM 1990 to present). In addition, experts in this area were asked to recommend extra reading materials. Additional references in textbooks and articles were examined. CONCLUSIONS: The literature review supports that there are major barriers to be addressed in the areas of availability, access, and utilization of preventive health care services for young children and their families. Major concerns include mandatory system for preventative health care, lack of health insurance coverage, cultural issues, and parental effects. IMPLICATIONS FOR PRACTICE: Health professionals in the community will need to work together to reevaluate current preventive health care practices for young children. Alternative methods for approaching and providing preventive health care services may become increasingly important if these services for young children are to be provided at current or increased levels. PMID- 11892537 TI - Preventing pediatric obesity: assessment and management in the primary care setting. AB - PURPOSE: To review the literature on and discuss the role of the primary care provider in assessing and managing overweight children before they become obese. DATA SOURCES: Selected research, national guidelines and recommendations, and the professional experience of the authors. CONCLUSIONS: The focus of primary care involves early detection and family interventions that are designed for lifestyle modifications, specifically for improved nutrition and an increase in regular physical activity, to achieve optimal child health. Early identification and management of children who exceed a healthy weight for height, gender, and age will prevent the increasing incidence of pediatric obesity. Early prevention and management of pediatric overweight and obesity will also decrease the potential for associated medical and psychosocial problems. IMPLICATIONS FOR PRACTICE: Pediatric obesity has risen dramatically in the United States during the last two decades; it is a significant child health problem that is preventable and largely under-diagnosed and under-treated. It is essential to discuss prevention of obesity with parents at every well-child visit; treatment should be initiated when patterns of weight gain exceed established percentiles for increasing height for age and gender. PMID- 11892539 TI - New vaccine technology--what do you need to know? AB - PURPOSE: To review the vaccine technology presently under development, in trials, or in some cases already available with a focus on applications for the primary care provider. DATA SOURCES: Selected scientific literature and Internet sources. CONCLUSIONS: Vaccine technology has made significant advances in the last 100 years. Newer approaches in vaccine designs include recombinant subunit vaccines, reassortment virus vaccines, live viral vectors, conjugate vaccines, and DNA vaccines. Traditional vaccines are generally expensive to produce and both live vaccines and protein-based vaccines require special storage conditions. These factors limit the worldwide usefulness of the familiar vaccines of today. IMPLICATIONS FOR PRACTICE: Keeping abreast of changes in this rapidly changing technology is necessary for health care providers to fulfill their roles effectively. New protocols that use nasal, vaginal, and intraperitoneal routes as well as subcutaneous injections and skin patches, edible vaccines and time release methods may be preferred depending on factors such as the age, health status, and emotional status of the patient. These advances hold promise for further progress in the development of vaccines that are protective against multiple organisms in more acceptable delivery systems. PMID- 11892540 TI - Pathogenesis and treatment of rhabdomyolysis. AB - PURPOSE: To provide clinicians in primary and acute care settings with information on the evaluation, diagnosis, and treatment of rhabdomyolysis. DATA SOURCES: Selected review articles from print and electronic sources, and a case study. CONCLUSIONS: Rhabdomyolysis is a common disorder, which may result from a large variety of diseases, trauma, or toxic insults to skeletal muscle. It is caused by skeletal muscle injury and results in the release of muscle contents (i.e., myoglobin) into the plasma. Muscular, urinary, and general internal disturbances are the three areas where signs and symptoms occur. IMPLICATIONS FOR PRACTICE: Clinical signs and symptoms vary widely. Associated life-threatening complications include acute renal failure, cardiac arrhythmias and arrest, disseminated intravascular coagulation, and compartment syndrome. Early recognition and treatment in the acute phase of rhabdomyolysis are keys to successful outcomes and an excellent recovery. PMID- 11892541 TI - Advanced practice nurses: a study of client satisfaction. AB - PURPOSE: To describe the satisfaction of clients with care provided by advanced practice nurses (APNs) in the Wright State University (WSU) Pilot Project. The Pilot Project was created through state legislation that recognized APNs associated with three universities in Ohio for the purpose of granting the APNs title protection, prescriptive authority, and recognition leading to reimbursement. DATA SOURCES: The Interactional Model of Client Health Behavior (Cox, 1982), which proposes that client characteristics and interactions with health professionals determine health outcomes, served as the framework for this descriptive study. The study measured client (n = 506) satisfaction with care delivered by 36 APNs at 26 different practice sites using an adapted Client Satisfaction Tool (CST). The internal consistency of the adapted CST was 0.935. CONCLUSIONS: Client (n = 506) satisfaction scores ranged from 28 to 50, with a mean score of 47.15 (SD = 4.00). Findings suggest that clients are very satisfied with APNs in the WSU Pilot Project. IMPLICATIONS FOR NURSING PRACTICE: Client satisfaction is one indicator used to determine quality of care. Consumer satisfaction benefits the individual client and provides important information for the health care payer. Measuring and reporting client satisfaction with care provided by APNs may increase the visibility and marketability of APNs. PMID- 11892542 TI - Factors influencing the access to prenatal care by Hispanic pregnant women. AB - PURPOSE: To explore factors influencing the access to prenatal care among Hispanic pregnant women living in the United States. DATA SOURCES: A convenience sample of 46 Hispanic migrant pregnant women was interviewed over a 12-month period using a set of five open-ended questions. CONCLUSIONS: The ability of the health care providers to communicate in Spanish, as well as the availability of culturally sensitive prenatal care were the main factors influencing the willingness of Hispanic women to access to prenatal care. IMPLICATIONS FOR PRACTICE: With the Hispanic population increasing in the United States, there is a need to provide culturally appropriate health care for that population; prenatal care is one of the areas of health care in demand. This study supports the findings in the literature and provides nurse practitioners a deeper understanding of the needs of Hispanic women needing prenatal care. PMID- 11892543 TI - Evaluation of a client care delivery model, Part 2: Variability in client outcomes in community home nursing. AB - Using similar variables, Part II explores variation in client outcomes such as OMAHA knowledge, behavior, and SF-36 scores. Medical and nursing diagnoses explained large variations in client outcomes. Clients cared for by degree prepared nurses improved knowledge and behavior scores. Unanticipated case complexity was negatively associated with client outcome even with nursing intervention. The study revealed that "for every unit increase in assignment of baccalaureate-prepared nurses, clients will on average demonstrate an 80% greater likelihood of improvement in knowledge scores and a 120% greater likelihood of improvement in behavior scores in relation to their health condition at discharge." This two-part study has offered insight into the controllable variables influencing the cost and quality of home care services. PMID- 11892544 TI - Case management: an evaluation at Childrens Hospital Los Angeles. AB - This prospective, quantitative, and qualitative evaluation of the case management program at CHLA clearly demonstrated the value of professional service coordination of care for children with complex, special health needs. Most specifically, the program documented improvement in three discrete areas of evaluation: 1. Financial, with decreased unnecessary expenditures and increased revenue. 2. Patient satisfaction, documented with validated questionnaires. 3. Clinical process improvement, using quantifiable clinical outcomes. At the very least, case management is an extremely valuable service in the present managed health care environment, and may in fact be indispensable. PMID- 11892545 TI - Capacity planning in hospital nursing: a model for minimum staff calculation. AB - An aging population, emerging technology, heightening patient expectations, rising health care costs, shorter patient stays, and growing pressure to improve quality have made the management of nursing resources even more critical today. While approaching a model for staffing levels, the authors considered factors such as patient acuity, work redesign, and minimum quality standards. The methodology for analysis included estimating the time needed to complete nursing tasks and calculating the average number of tasks per patient. With respect to nursing quality measures, the study examined the adequacy of nursing documentation including admission history, assessments, nursing procedures, and discharge report as well as nursing-driven outcomes such as fall and phlebitis rates. Lastly, the authors determined the theoretical number of staff needed to provide nursing care according to quality standards. PMID- 11892546 TI - A note on nurse self-scheduling. AB - Self-scheduling began in the 1960s and many hospitals have been using it with success. Benefits include saving management time, improved morale and professionalism, and cost reduction on matters related to personnel turnover. For nursing units experiencing scheduling challenges, self-scheduling may be a solution. PMID- 11892547 TI - Recent events highlight importance of mental health services. PMID- 11892548 TI - The leader as chief knowledge officer. AB - The most strategic asset a leader has to work with is the competency of her/his staff to perform at a level of excellence. The successful leader has the ability to create Communities of Practice within the organization that create highly bonded and effective communities of practice throughout the organization. PMID- 11892549 TI - Marketing and branding. AB - The advantages of marketing and branding nursing in a health care facility are discussed. Specific examples of marketing and branding are also described. PMID- 11892550 TI - The more things change. AB - The nursing profession's ability to influence public policy rests on individual as well as collective actions. Given Web-based information resources available today, nurses, regardless of where they reside, can be informed and engaged in health policy. PMID- 11892551 TI - The nursing shortage: can information technology help? AB - There is no doubt that the nursing shortage is a real and valid concern for the medical community. American Hospital Association President Dick Davidson reports: "America needs up to 126,000 nurses now, as well as more pharmacists, lab technicians, support staff and others. We need immediate action to help alleviate this situation" (American Hospital Association, 2001). Clinical information systems can be one of those immediate actions that can help nurses feel more confident about the care they are delivering. In addition to preventing medical errors, streamlining workflow and communications, and reducing redundant data entry, these systems can have a lasting and positive effect on overall job satisfaction, providing a significant influence on retaining our invaluable nursing resources. PMID- 11892552 TI - First, do no harm. PMID- 11892554 TI - Transplant service aims. PMID- 11892553 TI - In search of safety: an interview with Gina Pugliese. Interview by Alison P. Smith. AB - Gina Pugliese, MS, RN, is currently a vice-president for Premier, Inc. In addition to faculty and editorial positions, she directs international training courses in hospital epidemiology. She has made innumerable contributions to the field of safety over the past 25 years through her work with regulatory and federal agencies. Changing organizational culture from a "blame culture" to a "safety culture" is one notable challenge facing organizations. The airline industry has achieved significant changes in its safety culture by virtually eliminating a power hierarchy in the cockpit, a worthy example for operating rooms. Redesigning care delivery processes will minimize the opportunity for error if efforts are made to avoid reliance on memory, automate repetitive functions, reduce the number of steps involved, or use protocols when appropriate. The sophistication of consumers and the emergence of organizations like the Leap Frog Group will continue to challenge health care providers and potentially accelerate safety improvements. PMID- 11892555 TI - Packing arthrodesis wires. PMID- 11892556 TI - Formulating a policy for the returning of implanted prostheses. PMID- 11892557 TI - Electronic health records. PMID- 11892558 TI - Implant 'cannot wear out'. PMID- 11892559 TI - Developing a systematic review. Trials and tribulations. PMID- 11892560 TI - The role of an acute pain service. An overview. PMID- 11892561 TI - Influencing without authority. PMID- 11892562 TI - Surgeon wants nurse assistants. PMID- 11892563 TI - Wherefore N77? PMID- 11892564 TI - Recognised standard for the level of staffing required in the operating theatre to maintain a safe environment for the patient. PMID- 11892565 TI - Nurse autonomy. Pain control and discharge from recovery. PMID- 11892566 TI - Team-based self-rostering. PMID- 11892567 TI - Common head injuries, their diagnosis and treatment. PMID- 11892568 TI - Perioperative nurses in Uganda aid effort. PMID- 11892569 TI - Disposal of foetal remains. PMID- 11892570 TI - Artificial nails should not be worn in the operating room theatre. PMID- 11892571 TI - Defining theatre nursing. PMID- 11892572 TI - Interactive digital television. PMID- 11892573 TI - Accidents don't just happen! PMID- 11892574 TI - Career breaks. PMID- 11892575 TI - My story. Surviving a neurological accident. PMID- 11892576 TI - Nurse education in the new millennium. A review of current reports. PMID- 11892577 TI - The physiology of wound healing and wound assessment. AB - Wound healing is a complex and remarkable process that occurs when the body responds to trauma. It requires the interaction of several intricate processes with the ultimate aim being to maintain homeostasis. With advances in surgery and increases in the amount of surgery being undertaken, research into wound healing has become of great importance. Providing the optimum conditions for wounds to heal is of great significance as pressures on nursing and hospital resources become ever greater. PMID- 11892579 TI - Prioritising documentation. PMID- 11892578 TI - Oxygen reduces post-op infections. PMID- 11892580 TI - During the recent warm weather we have been experiencing problems with the regulation of temperature and humidity within our theatre suite. PMID- 11892581 TI - Reduced hours and flexitime. PMID- 11892582 TI - An analysis of perioperative care. AB - Perioperative nursing requires an appreciation of patients' needs throughout the episode of care. This article presents a critical analysis and examination of the perioperative care of a patient undergoing total abdominal hysterectomy. In addition to analysing the efficacy of care delivery in the perioperative setting, we will explore the application of nursing theory and research to everyday practice and the degree to which a hospital's infrastructure facilitates quality care. PMID- 11892583 TI - Burr holes. AB - Neurosurgical procedures may sometimes take place in general operating theatres, usually when a patient has sustained a life-threatening head injury. Burr holes may be made in accident and emergency departments before patients are transferred for further treatment. For these reasons it is useful for perioperative nurses in general hospitals to have some knowledge of neurosurgical operating techniques and instrumentation. PMID- 11892584 TI - Little cogs in a big wheel. PMID- 11892585 TI - Laryngospasm--touch is not the answer. PMID- 11892586 TI - Sensitivity or what? PMID- 11892587 TI - Who actually has responsibility for controlled drugs in the perioperative environment. PMID- 11892588 TI - Lies, damned lies and statistics. AB - Readers may remember Jim Miller's introductory article in this series (Miller 2001), where lessons learnt from the Titanic disaster were related to lessons to be learnt in the NHS today. Unfortunately many of those lessons are yet to be learnt by some of our organisations or, certainly, lessons which need to be applied consistently. One of those lessons related to the need for accurate data and using it to inform action and priorities. This article identifies some of the data requirements. where they might be obtained and the need for a systematic process to inform strategy. PMID- 11892589 TI - Caring for the carers. AB - In this further article of the series, Claire Campbell introduces and explores the needs of Health Service employees who care for their own relatives or companions who cannot cope due to sickness, disability or age, why it is important to recognise their needs, what kind of support they need, and how policies can be put in place. PMID- 11892590 TI - Electrosurgical smoke plume. Is it harmful to staff? AB - When a potential problem was identified by several staff regarding electrosurgical smoke plume, Angharad Hughes and Jonathan Hughes decided to look into the situation using an evidence-based approach. What follows is a precis of literature found, and information about how this approach was carried out. It shows that there is some evidence that particles present in smoke plume are potentially harmful to health, but that the extent of the risk is as yet unknown. PMID- 11892591 TI - How well do we use our resources? AB - We have already considered that the resources we have to work with in our organisations are not infinite. The most valuable of these are our people and our time. In addition, some patient work is not 'cost effective'. As technology and treatment options become more and more sophisticated, our overall fixed and variable costs can increase alarmingly. In this fifth article in the series, Fiona Westwood says we need, therefore, to look at all of our resources to maximise the value that they can bring. PMID- 11892592 TI - Healthcare support workers and the scrubbed role. AB - The scrubbed role is highly valued amongst many theatre nurses. A survey by Roberts (1989) found that 61.9% of 147 nurses stated that they preferred the scrubbed role to an anaesthetic role (4.8%). There is more recent evidence that nursing still dominates the scrubbed role, with over 50% of 108 nurses surveyed stating that they only worked in the scrubbed area of the operating department (Hind 1997). PMID- 11892593 TI - Know your heart. AB - In this article in our Concepts in Anatomy series, Mriga Williams joins John Clancy and Andrew McVicar in an examination of the heart, identifying and describing its component parts, its relationship to the circulatory system and some of the abnormalities it might develop. The article includes a range of activities readers may carry out to test and improve their knowledge. The series is based on the text by John Clancy and Andrew McVicar, Physiology and Anatomy--a homeostatic approach, Arnold, London (in print September 2001). PMID- 11892594 TI - A new lens. Through a colleague's eye. AB - Just like other members of the wider community, medical practitioners experience disorders that require corrective surgery. Michael Matthews has been retired for more than 18 years. At one time he was in charge of a coronary care unit. Recently, the tables were turned on him, when he had to go back into hospital, this time as a patient for day surgery to remove a cataract and receive a replacement lens. Dr Matthews describes his experience of eye problems and of receiving his new lens. PMID- 11892595 TI - NCMEC (National Center for Missing & Exploited Children): healthcare doing good job in guarding newborns, but watch out for complacency. PMID- 11892596 TI - How security directors can become more conversant with computer network security. PMID- 11892597 TI - The North Shore University Hospital security survey: from recommendations to reality. AB - A unique case history on how to use an outside consultant to conduct an extensive security survey and how to go about successfully implementing the resulting recommendations, is described in detail in this report. A project of the 731-bed North Shore University Hospital, Manhasset, Long Island, NY, the survey and its implementation is described by Chris Leibfried, CHPA, CSE, director of campus support services. Leibfried also is chairperson of the Security Directors Committee for the 18-hospital North Shore-Long Island Jewish Health System that covers the counties of Nassau, Suffolk, Queens, and Staten Island. PMID- 11892598 TI - Hospital keeps officer standards high despite recruiting difficulties. PMID- 11892599 TI - Keil: JCAHO likely to revise emergency preparedness requirements for hospitals. PMID- 11892600 TI - [50th anniversary volume of the Japanese Journal of Anesthesiology]. PMID- 11892601 TI - Nutrition, Diet and Oral Health for the 21st Century. Proceedings of the Public Health Section of an FDI/ILSI Europe Satellite Symposium. Paris, France, 28 November 2000. PMID- 11892602 TI - Nutrition, diet and oral health in the 21st century. PMID- 11892603 TI - A lifetime of accomplishment. PMID- 11892604 TI - The journey of dentistry. My journey from practitioner to educator. PMID- 11892606 TI - A passive-marker-based optical system for computer-aided surgery in otorhinolaryngology: development and first clinical experiences. PMID- 11892605 TI - Former dental hygienist has independent private practice. PMID- 11892607 TI - Prospective, randomized outcome study of endoscopy versus modified barium swallow in patients with dysphagia. PMID- 11892609 TI - Use of tenofovir. PMID- 11892608 TI - FDA approves tenofovir. PMID- 11892610 TI - Vaccine five years away. PMID- 11892611 TI - Positive partners slow to tell others. PMID- 11892612 TI - Hospitals slow with compliance. PMID- 11892613 TI - [The mystery of a red buttock]. PMID- 11892614 TI - The influence of the mixing technique on the content of voids in two polyether impression materials. AB - One of the most critical problems related to hand mixing of an elastomeric impression material is air entrapment during spatulation. This leads the formation of both surface and subsurface bubbles which in turn may result in inaccurate dental impressions and/or jeopardize their physical properties. In the present study the influence of the mixing technique has been determined by evaluating the surface area and the number of voids in two polyether materials (Permadyne and Impregum, Espe, Seefeld, Germany). The techniques tested were the stropping technique as hand-mixing and the Pentamix device (Espe) as mechanical mixing. Eighty special trays (10 mm x20 mm x43mm) featuring 10 transverse slots were fabricated and divided into four groups of twenty units. Groups 1 and 2 received the hand-mixed materials Permadyne high viscosity and Impregum F, respectively. Groups 3 and 4 (Pentamix group) received the mechanically mixed materials Permadyne Penta H and Impregum Penta, respectively. After polymerization, 10 slices of material were obtained for each tray by sectioning through the tray slots with a surgical blade. The slices were glued on a card and black and white photographs were taken. Subsequently, the negative films were placed on a viewing box and digitized with a video camera. A special software program allowed to identify and calculate the total surface area and the number of voids. Significant differences between the "stropping" groups (groups 1 and 2) and "Pentamix" groups (groups 3 and 4) were found. The mechanical mixing (Pentamix) generated the smallest number and total surface area of voids, while no significant differences were detected between Permadyne Penta H and Impregum Penta. Clearly, mechanical mixing represents a marked improvement over the traditional hand-mixing methods. PMID- 11892615 TI - Fluid balance documentation: a case study of daily weight and intake/output omissions. AB - Daily intake and output (I&O) and weight documentation is frequently incomplete or omitted completely, even when a physician's order is in place. This brief case study illustrates the importance of maintaining these records to avoid fluid balance disorders and negative patient outcomes, as well as support patient well being. PMID- 11892616 TI - Suppose feds don't depose. HCA seeks to prevent employee testimony. PMID- 11892617 TI - The right prescription. Health system's medical practice will turn profit despite hurdles. PMID- 11892618 TI - Health Care Hall of Fame. Legendary leadership. Three new inductees left mark on healthcare. PMID- 11892619 TI - Steamrolling ahead. Two N.Y. hospitals don't let a little paperwork get in the way. PMID- 11892620 TI - Kansas nixes Anthem-Blues deal. PMID- 11892621 TI - Quietly working on quality. Hospitals, aided by associations are addressing errors. PMID- 11892622 TI - Quality is a mandate. AHA task force latest bid to improve patient safety. PMID- 11892624 TI - Utah could open the floodgates. PMID- 11892625 TI - Research governance framework for health and social care. PMID- 11892623 TI - Looking for a breakthrough. Premier's hospitals collaborate on quality initiatives. PMID- 11892626 TI - Bibliography. Current world literature. Hematology and oncology. PMID- 11892627 TI - Bibliography. Current world literature. Orthopedics. PMID- 11892629 TI - New website highlights HIV resources. PMID- 11892628 TI - Bibliography. Current world literature. Infectious diseases and immunization. PMID- 11892630 TI - Study looks at HCV therapy responses. PMID- 11892631 TI - Combination resistance test available. PMID- 11892632 TI - TMC125 shows promise. PMID- 11892633 TI - Chromosomal imbalances in primary oligodendroglial tumors and their recurrences: clues about malignant progression detected using comparative genomic hybridization. AB - OBJECT: Despite the rapid increase in knowledge concerning the genetic basis of malignant progression in astrocytic tumors, progression of oligodendroglial tumors (including both pure oligodendrogliomas and mixed oligoastrocytomas) is still poorly understood. The aim of the present study is the elucidation of chromosomal imbalances involved in the progression of oligodendroglial tumors toward malignancy. METHODS: Using comparative genomic hybridization (CGH) on snap frozen tumor tissue, the tumor genomes of five primary oligodendroglial tumors and associated recurrent tumors were screened for chromosomal imbalances. This information was correlated with clinical data (including follow-up data) and histopathological malignancy grade. In all cases an increase in chromosomal imbalances was detected in the recurrent tumor, indicating genetic progression. In three of the five cases this correlated with malignant progression detected at the histopathological level. The results indicate that, similar to what occurs in astrocytic tumors, chromosomal imbalances harboring genes involved in the cell proliferation control mechanism at the G1-S border are involved in the progression of oligodendroglial tumors. Additionally, although gains of genetic material on chromosome 7 and losses on chromosome 10 are most frequently detected in the course of malignant progression of astrocytic tumors, either or both of these can also occur during malignant progression of typical oligodendroglial tumors that contain losses involving chromosome 1p and/or chromosome 19q. CONCLUSIONS: When performed on optimally preserved material from a small set of primary oligodendroglial tumors and associated recurrent tumors, CGH detects chromosomal aberrations that potentially play a mechanistic role in the malignant progression of these tumors. PMID- 11892634 TI - Acute distortion of the anatomy of the third ventricle during third ventriculostomy. Report of four cases. AB - The authors report on four third ventriculostomy procedures in which upward ballooning of the third ventricular floor occurred immediately after perforation of the floor and withdrawal of a Fogarty catheter. The floor herniated into the third ventricle, hindering the endoscopic view. Preoperative magnetic resonance imaging demonstrated a similar anatomy in all four cases, consisting of hydrocephalus, extreme dilation of the third ventricle, and disappearance of the interpeduncular cistern due to a very thin, membranous floor of the third ventricle, which herniated downward, draping over the basilar artery. The authors suggest that excessive rinsing in combination with this anatomical configuration provoked the phenomenon of upward ballooning of the third ventricular floor, which is described in this report. PMID- 11892635 TI - Patient-controlled epidural fentanyl following spinal fentanyl at Caesarean section. AB - Spinal fentanyl can improve analgesia during Caesarean section. However, there is evidence that, following its relatively short-lived analgesic effect, there is a more prolonged spinal opioid tolerance effect. The effectiveness of postoperative epidural fentanyl analgesia may therefore be reduced following the use of spinal fentanyl at operation. This randomised, double-blind study was designed to assess whether patient-controlled epidural fentanyl could produce effective analgesia following 25 microg of spinal fentanyl at operation. Patients undergoing elective Caesarean section received spinal bupivacaine combined with either fentanyl 25 microg (fentanyl group; n = 18) or normal saline (saline group; n = 18). Patient controlled epidural fentanyl was used for postoperative analgesia. The fentanyl group used a mean of 23.4 (SD 14.5) microg x h(-1) of fentanyl, compared with 27.0 (10.8) microg x h(-1) for the saline group (p =0.41). Using a 0-100 mm visual analogue score for pain, the maximum pain score recorded at rest for the fentanyl group was median 24 [IQR 15-35] mm, compared with 15 [13-45] mm for the saline group (p = 0.41). The maximum pain score recorded on coughing for the fentanyl group was 29 [24-46] mm, compared with 27 [19-47] mm for the saline group (p = 0.44). Nine of the fentanyl group rated postoperative analgesia as excellent and nine as good, compared with 10 of the saline group who rated it as excellent and eight as good (p = 0.74). Epidural fentanyl can produce effective analgesia following the use of 25 microg spinal fentanyl at Caesarean section. PMID- 11892636 TI - The management of pain following day-case surgery. AB - The object of this study was to assess patients' experience of pain management following day surgery. One hundred and two patients agreed to take part in a telephone survey, 2 and 4 days following day surgery. The majority of patients (73%) were broadly satisfied with the quality of pain management they received, however, there was room for improvement. Despite modern anaesthesia and surgery, 17% of patients surveyed reported having severe pain immediately following day case surgery. The majority (82%) of patients left the day-case ward in pain and an even higher proportion (88%) had pain at some time between 2 and 4 days postoperatively. Severe levels of pain following discharge from hospital were a concern for 21% of patients. It was reported that day-case staff did not always ask patients whether they were in pain. Communication with patients is vital in the delivery of optimal care. More support and more information are needed to manage patients' pain effectively, whilst in the day-case wards and also following discharge, at home. PMID- 11892637 TI - Measurement of tracheal tube cuff pressure in critical care. AB - We conducted an observational study to measure tracheal tube cuff pressures in the critical care environment, where prolonged intubation is common. Thirty-two patients were studied. Sixty-two per cent of all tracheal cuffs had intracuff pressures above the recommended value. We also conducted a telephone survey of 24 intensive care units within the Northern and Yorkshire Region, which showed that 75% of the intensive care units never checked tracheal tube cuff pressures. Critically ill patients are particularly vulnerable to tracheal injury due to prolonged intubation. We suggest that cuff pressures should be measured regularly. PMID- 11892638 TI - An evaluation of ultrasound imaging for identification of lumbar intervertebral level. AB - The accuracy of ultrasound imaging to identify lumbar intervertebral level was assessed in 50 patients undergoing X-ray of the lumbar spine. Using an ultraviolet marker, an anaesthetist attempted to mark the L2/3, L3/4 and L4/5 intervertebral spaces. A radiologist unaware of these marks attempted to mark the same spaces with the aid of ultrasound imaging. X-ray-visible pellets were taped to the back at the various marks prior to lateral lumbar X-ray. Ultrasound imaging identified the correct level in up to 71% of cases, but palpation was successful in only 30% (p < 0.001). Up to 27% of marks using the palpation method were more than one spinal level above or below the assumed level using palpation, but none were more than one level high or low using ultrasound guidance. PMID- 11892639 TI - The effect of swabs soaked in bupivacaine and epinephrine for pain relief following simple dental extractions in children. AB - We studied 133, ASA I or II children, aged 5-12 years undergoing general anaesthesia for simple dental extractions. Induction and maintenance of anaesthesia were achieved using sevoflurane in nitrous oxide and oxygen. At the end of surgery, patients had swabs soaked in a trial solution placed over the exposed teeth sockets. The bupivacaine group had swabs soaked in bupivacaine 0.25% with epinephrine 1:200 000, the saline group had swabs soaked in saline. Pain scores were recorded on a 4-point scale as follows: 0 = I don't hurt at all; 1 = I hurt a bit; 2 = I hurt a lot; 3 = I hurt the most. Nurse pain assessments and the patient's own scores were recorded at 15 and 30 min following recovery from anaesthesia. The median pain scores (2 at 15 min and 1 at 30 min postoperatively) were the same in both groups. PMID- 11892640 TI - Disposable laryngoscope blades. PMID- 11892641 TI - Overfilling of vaporisers. PMID- 11892642 TI - The Pro-Seal laryngeal mask airway. PMID- 11892643 TI - Gum elastic bougies. PMID- 11892644 TI - Call for an alternative coupling system for regional equipment. PMID- 11892645 TI - Obesity and day-case surgery in an isolated unit. PMID- 11892646 TI - Evaluation of the Pneupac Ventipac portable ventilator in critically ill patients 1. PMID- 11892647 TI - Evaluation of the Pneupac Ventipac portable ventilator in critically ill patients 2. PMID- 11892648 TI - Aberrant brachio-cephalic artery precluding placement of tracheostomy. PMID- 11892649 TI - Anaesthesia for Stiff-Person Syndrome. PMID- 11892650 TI - Report of vaporiser malfunction. PMID- 11892651 TI - Up-and-down allocation. PMID- 11892652 TI - Wound botulism in injecting drug users. PMID- 11892653 TI - Not quite crystal clear. PMID- 11892654 TI - Hypotension prophylaxis for Ceasarean section. PMID- 11892655 TI - Epidural catheter. PMID- 11892656 TI - Aspirin and central neural blockade. PMID- 11892657 TI - An audit of the use of low molecular weight heparin and epidural anaesthesia. PMID- 11892658 TI - A simple technique to reduce the incidence of accidental dural puncture. PMID- 11892659 TI - Transient neurological manifestations after epidural analgesia with ropivacaine. PMID- 11892660 TI - Intrathecal air following spinal anaesthesia. PMID- 11892661 TI - Pre-operative fluid fasting for adult elective surgery. PMID- 11892662 TI - Recording the confirmation of nasogastric tube placement. PMID- 11892663 TI - A new angle on a Tuohy needle. PMID- 11892664 TI - A possible cause of corneal abrasions. PMID- 11892665 TI - A faulty Yankauer suction catheter. PMID- 11892666 TI - The significance of post mortem tryptase levels in supporting a diagnosis of anaphylaxis. PMID- 11892667 TI - Audit of audit ... PMID- 11892668 TI - Radial artery cannulation - a simple positioning aid. PMID- 11892669 TI - Blocked emergency syringe. PMID- 11892670 TI - Malignant hyperpyrexia and a career in Anaesthesia. PMID- 11892671 TI - e-Surveys. PMID- 11892672 TI - Better skin incision for Seldinger dilator insertion. PMID- 11892674 TI - All in the name of progress. PMID- 11892673 TI - Another use for the laryngoscope. PMID- 11892676 TI - Genetic parameters for various random regression models to describe the weight data of pigs. AB - Various random regression models have been advocated for the fitting of covariance structures. It was suggested that a spline model would fit better to weight data than a random regression model that utilizes orthogonal polynomials. The objective of this study was to investigate which kind of random regression model fits best to weight data of pigs. Two random regression models that described weight of individual pigs, one using orthogonal polynomials, and the other using splines, were compared. A comparison with a multivariate model, Akaike's information criterion, and the Bayesian-Schwarz information criterion were used to select the best model. Genetic, permanent environmental, and total variances increased with age. Heritabilities for the multivariate model ranged from 0.14 to 0.19, and for both random regression models the heritabilities were fluctuating around 0.17. Both genetic and phenotypic correlations decreased when the interval between measurements increased. The spline model needed fewer parameters than the multivariate and polynomial models. Akaike's information criterion was least for the spline model and greatest for the multivariate model. The Bayesian-Schwarz information criterion was least for the polynomial model and greatest for the multivariate model. Residuals of all models were normally distributed. Based on these results, it is concluded that random regression models provide the best fit to pig weight data. PMID- 11892677 TI - Comparison of models for estimation of genetic parameters for mature weight of Hereford cattle. AB - Genetic parameters of mature weight are needed for effective selection and genetic evaluation. Data for estimating these parameters were collected from 1963 to 1985 and consisted of 32,018 mature weight records of 4,175 Hereford cows that were in one control and three selection lines that had been selected for weaning weight, for yearling weight, or for an index combining yearling weight and muscle score for 22 yr. Several models and subsets of the data were considered. The mature weight records consisted of a maximum of three seasonal weights taken each year, at brand clipping (February and March), before breeding (May and June), and at palpation (August and September). Heritability estimates were high (0.49 to 0.86) for all models considered, which suggests that selection to change mature weight could be effective. The model that best fit the data included maternal genetic and maternal permanent environmental effects in addition to direct genetic and direct permanent environmental effects. Estimates of direct heritability with this model ranged from 0.53 to 0.79, estimates of maternal heritability ranged from 0.09 to 0.21, and estimates of the genetic correlation between direct and maternal effects ranged from -0.16 to -0.67 for subsets of the data based on time of year that mature weight was measured. For the same subsets, estimates of the proportions of variance due to direct permanent environment and maternal permanent environment ranged from 0.00 to 0.09 and 0.00 to 0.06, respectively. Using a similar model that combined all records and included an added fixed effect of season of measurement of mature weight, direct heritability, maternal heritability, genetic correlation between direct and maternal effects, proportion of variance due to direct permanent environmental effects, and proportion of variance due to maternal permanent environmental effects were estimated to be 0.69, 0.13, -0.65, 0.00, and 0.04, respectively. Mature weight is a highly heritable trait that could be included in selection programs and maternal effects should not be ignored when analyzing mature weight data. PMID- 11892678 TI - Correlations among selected pork quality traits. AB - Establishing relationships among specific quality traits is important if significant progress toward developing improved pork quality is to be realized. As part of a study to examine the individual effects of genes on meat quality traits in pigs, a three-generation resource family was developed. Two Berkshire sires and nine Yorkshire dams were used to produce nine F1 litters. Sixty-five matings were made from the F1 litters to produce four sets of F2 offspring, for a total of 525 F2 animals used in the study. These F2 animals were slaughtered at a commercial facility upon reaching approximately 110 kg. Carcass composition traits, pH measurements, and subjective quality scores were made at 24 h postmortem. Loin samples (n = 525) were collected at 48 h postmortem, and meat quality traits were evaluated. These traits included pH (48 h), Hunter L-values, drip loss, glycolytic potential, ratio of type IIa/IIb myosin heavy chains (IIa/IIb), total lipid, instrumental measures of tenderness using the Star Probe attachment of the Instron, cook loss measurements, and sensory evaluations. Significant phenotypic correlations were found between many carcass, instrumental, and biochemical measurements, and sensory quality traits. Star Probe measurements were significantly correlated with drip loss (0.29), glycolytic potential (0.30), pH (-0.29), total lipid (-0.14), and Hunter L-values (0.28). Drip loss was significantly correlated with glycolytic potential (0.36), pH (-0.28), IIa/IIb (-0.10), and Hunter L-values (0.33). Hunter L-values were also significantly correlated with total lipid (0.33) and IIa/ IIb (-0.11). Sensory tenderness, flavor, and off-flavor scores were significantly correlated with drip loss, pH, and glycolytic potential measurements. Marbling score, total lipid, and drip loss were not significantly correlated with sensory juiciness scores, but cooking loss was. Marbling and total lipid were significantly correlated with firmness scores (0.37 and 0.31, respectively). Taken together, the data in this study suggest that changes in some meat quality traits can affect many other meat quality attributes. The correlations yield information that could aid in directing future studies aimed at understanding the underlying biological mechanisms behind the development of many quality traits. PMID- 11892679 TI - The effects of potassium diformate and its molecular constituents on the apparent ileal and fecal digestibility and retention of nutrients in growing-finishing pigs. AB - Five 43-kg barrows [(Dutch Landrace x Yorkshire) x Yorkshire] were fitted with steered ileocecal valve cannulas to compare the effects of K-diformate (KDF), a specifically conjugated salt vs its molecular constituents, namely, formic acid and K-formate, as acidifiers in lysine-deficient diets on the apparent ileal (ID) and fecal digestibility, retention of nutrients, and manure production. The animals were randomly assigned to five dietary treatments according to a 5 x 5 Latin square design as follows: 1) control-no acidifier; 2) 1% KDF (= 0.65% K formate + 0.35% formic acid, or 0.7% [HCOO-] + 0.3% [K+]); 3) 0.65% K-formate (= 0.35% [HCOO-] + 0.3% [K+]); 4) 0.35% formic acid (= 0.35% [HCOO-]); and 5) 1.3% K formate (= 0.7% [HCOO-] + 0.6% [K+]). Diets were formulated with barley, wheat, soybean meal, and canola meal as major ingredients, and provided all nutrients at adequate levels, except for lysine (24% less than estimated requirement). Feeding level was equal to 2.5 x maintenance requirement (MR) for ME (MR = 418 kJ ME x BW(-0.75)), and daily rations were given in two portions after mixing with water in a ratio of 1:2.5. Chromic oxide was used as an indigestible marker. No clinical health problems due to the dietary treatments were observed. Irrespective of the additive, there were no differences (P < or = 0.10) in the ID of DM, OM, CP, or essential amino acids compared to the control, except for phenylalanine (P < or = 0.05). Among nonessential AA, only the ID of tyrosine tended (P = 0.092) to increase (up to 3.9 percentage units). The fecal digestibility of ash and K were greater (P < or = 0.001) in pigs fed supplemental K, irrespective of its source. The greater intake and fecal digestibility of K corresponded with greater (P < or = 0.05) losses of K in urine. Body retention of N, Ca, total P, and K was similar (P > or = 0.10) among treatments. As estimated from a separate nonorthogonal analysis, supplemental K improved (P < or = 0.05) body N by 3.7 percentage units compared to the control. The results of this study do not provide a clear explanation for the improved growth performance reported previously with KDF and its molecular constituents, and further research on their in vivo mode of action will require methodological refinement, especially with regard to the efficiency of AA utilization. PMID- 11892680 TI - Rapid communication: physical mapping of two bovine microsatellite loci. PMID- 11892682 TI - [The 102nd meeting of the Japan Surgical Society. Kyoto, Japan. April 11-13, 2001. Program]. PMID- 11892681 TI - Office-based transurethral microwave thermotherapy using the TherMatrx TMx-2000. AB - BACKGROUND AND PURPOSE: Transurethral microwave thermotherapy (TUMT) is an effective therapy for symptomatic benign prostatic hyperplasia (BPH), but the trade-off between the magnitude of clinical improvement and side effects and patient tolerance has limited its appeal to patients and urologists. This study, using the TherMatrx TMx-2000, a TUMT device that directly heats the transition zone to greater than 50 degrees C, has been focused on resolving these issues and developing a truly office-based therapy that is well tolerated with a benign post treatment course. PATIENTS AND METHODS: This study was multi-institutional and designed as a blinded, randomized, and sham-controlled trial. A series of 200 patients with an AUA Symptom Index (AUASI) of >12, a peak flow rate of <12 mL/sec, and cystoscopic evidence of BPH were randomized 2:1 (active to sham) and treated in seven physician offices under a Food and Drug Administration supervised and audited premarket approval protocol. No intravenous sedation was used in any patient. Follow-up for the sham-treatment group was 3 months, at which time, patients could cross over to an active treatment. A total of 119 patients have completed 1-year follow-up. RESULTS: The active and sham groups were statistically identical at baseline. The 1-hour total treatment was extremely well tolerated using urethral lidocaine and oral medications; not a single prostate block or parental dose of medication was required. The active treatment group demonstrated a statistically significant reduction (p < 0.05) in AUASI at 3 months compared with sham treatment, with an AUASI decrease from 22.4 to 12.4 (n = 124) for active v 22.9 to 17 for sham (n = 62). For the 119 patients in the active arm who have reached 12 months, the AUASI has fallen to 10.6 points (47.1% decrease), and the peak flow rate has increased 5.0 mL/sec (58.1%). Postprocedure catheterization was typically 2 or 3 days, and the 16.8% of patients who failed their first voiding trial all voided within 1 week. No major adverse events such as stricture, rectal findings, or ejaculatory changes have been reported. CONCLUSIONS: This study demonstrates that the TherMatrx TMx-2000 TUMT effectively treats symptomatic BPH in the physician office with minimal morbidity. PMID- 11892683 TI - The role of bacterial toxins in sudden infant death syndrome (SIDS). AB - There is increasing evidence for the involvement of bacterial toxins in some cases of sudden infant death syndrome (SIDS), particularly the pyrogenic toxins of Staphylococcus aureus. This had led to the hypothesis that some SIDS deaths are due to induction of inflammatory mediators by infectious agents or their products during a period in which the infant is unable to control these normally protective responses. The genetic, developmental and environmental risk factors identified for SIDS are assessed in relation to frequency or density of mucosal colonisation by toxigenic bacteria and their effects on induction and control of inflammatory responses to the toxins. PMID- 11892684 TI - Effects of the adjuvant cholera toxin on dendritic cells: stimulatory and inhibitory signals that result in the amplification of immune responses. AB - Cholera toxin (CT) is a potent mucosal adjuvant. When administered through the mucosal route CT amplifies B and T lymphocyte responses to co-administered antigens. Since the discovery of CT as a mucosal adjuvant, other bacterial enterotoxins have been found to have this property. These molecules or their detoxified derivatives are all important for the development of mucosal vaccines for human use, and it is thus necessary to understand their mechanism of action. CT has immunomodulatory effects on different cell types, however, the interaction of CT with dendritic cells (DCs), which have a primary role in the priming of immune responses, may be crucial for its adjuvant activity. PMID- 11892685 TI - Use of Arnica to relieve pain after carpal-tunnel release surgery. AB - CONTEXT: Arnica is commonly used by the public as a treatment for bruising and swelling. OBJECTIVE: To assess whether Arnica administration affects recovery from hand surgery. DESIGN: Double-blind, randomized comparison of Arnica administration versus placebo. SETTING: Specialist hand surgery unit at the Queen Victoria NHS Trust. PARTICIPANTS: Thirty-seven patients undergoing bilateral endoscopic carpal-tunnel release between June 1998 and January 2000. INTERVENTION: Homeopathic Arnica tablets and herbal Arnica ointment compared to placebos. MAIN OUTCOME MEASURES: Grip strength, wrist circumference, and perceived pain measured 1 and 2 weeks after surgery. RESULTS: No difference in grip strength or wrist circumference was found between the 2 groups. However, there was a significant reduction in pain experienced after 2 weeks in the Arnica treated group (P<.03). CONCLUSIONS: The role of homeopathic and herbal agents for recovery after surgery merits further investigation. PMID- 11892686 TI - Measuring treatment effects of cilostazol on clinical trial endpoints in patients with intermittent claudication. AB - Intermittent claudication (IC) comprises the most common presenting symptoms of peripheral arterial disease (PAD), which itself is a manifestation of systemic atherosclerosis. Typical symptoms of IC are aching pain, numbness, and fatigue in the lower extremities. Symptoms are induced by walking or exercise and usually resolve with rest. The cornerstone of treating IC is risk-factor reduction and a supervised exercise regimen. Pharmacotherapy specifically indicated for the treatment of IC includes a new drug, cilostazol, and the traditional drug, pentoxifylline. Cilostazol also has antiplatelet, antithrombotic, and vasodilatory activity, as well as a positive effect on serum lipids. Eight multicenter clinical trials, seven in the U.S. and one in the U.K., used objective and subjective clinical endpoints to assess the treatment efficacy of cilostazol. Objective endpoints included maximal and pain-free walking distance (MWD and PFWD, respectively), the ankle-brachial index, peripheral hemodynamic measurements, and serum lipid levels. Subjective endpoints, assessed by patient questionnaires, included perceived functional status and health-related quality of life. Cilostazol treatment showed statistically significant increases in MWD and PFWD within 4 weeks, as well as improvements in physical functional status at 24 weeks, compared with placebo and pentoxifylline. Increases in high-density lipoprotein cholesterol and decreases in plasma triglycerides were also noted. Subjective assessments appeared to match objective parameters. PMID- 11892687 TI - Evidence for recombination among the tomato leaf curl virus strains/species from Bangalore, India. AB - Tomato leaf curl virus (ToLCV) belongs to the Begomovirus genus of the family Geminiviridae. These viruses have circular single stranded DNA molecules as their genome encapsidated in icosahedral geminate particles. Generally the Begomoviruses are bipartite with respect to their genomic composition. ToLCV from South India is unique in that only DNA A component has been isolated and sequenced thus far and there is no evidence for the presence of DNA B component. In this communication we report the genomic sequences of DNA A component of two strains of Tomato leaf curl Bangalore virus (ToLCBV), one from Bangalore, ToLCBV [Ban 5] and the other from Kolar (70 kms from Bangalore), ToLCBV [Kolar]. We have examined the possibility of recombination between strains/species that co-exist within the same geographical location. A novel method has been used to analyze the variation of ToLCV sequences reported from Bangalore and to assess the frequency and importance of recombinational events among the strains/species existing in Bangalore. The results indicate that there are potential sites of recombination in AV1, AV2, AC1 and intergenic regions of the viral genome and this accounts for the observed variability in these strains/species. PMID- 11892688 TI - Presence of European bat lyssavirus RNas in apparently healthy Rousettus aegyptiacus bats. AB - Apparently healthy Rousettus aegyptiacus bats were randomly chosen from a Dutch colony naturally infected with European bat lyssavirus subgenotype 1a (EBL1a). These bats were euthanised three months after the first evidence of an EBL1a infection in the colony. EBL1a genomic and antigenomic RNAs of the nucleoprotein gene were detected by nested reverse transcriptase PCR in 75% of the examined Rousettus aegyptiacus bats. The EBL1a RNAs of the nucleoprotein gene were detected mainly in brain tissues, but also in other organs. EBL1a messenger RNAs of the nucleoprotein gene and the glycoprotein gene were detected in brain tissues. The standard fluorescent antibody test revealed the presence of lyssavirus antigens in brain tissues from 7 (17.5%) Rousettus aegyptiacus bats. Furthermore, EBL1a could not be detected by virus isolation on murine neuroblastoma cells or by intracerebral inoculation of suckling mice. Neutralising antibodies directed against EBL1 were detected in 11% of the examined bats. This study shows that at least 85% of the apparently healthy Rousettus aegyptiacus bats must have been infected with EBL1a, and that these bats may survive from an EBL1a infection. Furthermore, the study supports the possibility of a long-term maintenance of EBL1a genome in Rousettus aegyptiacus bats. PMID- 11892689 TI - Modeling and real-time prediction of classical swine fever epidemics. AB - We propose a new method to analyze outbreak data of an infectious disease such as classical swine fever. The underlying model is a two-type branching process. It is used to deduce information concerning the epidemic from detected cases. In particular, the method leads to prediction of the future course of the epidemic and hence can be used as a basis for control policy decisions. We test the model with data from the large 1997-1998 classical swine fever epidemic in The Netherlands. It turns out that our results are in good agreement with the data. PMID- 11892690 TI - A unified parametric regression model for recapture studies with random removals in continuous time. AB - Conditional likelihood based on counting processes are combined with a Horvitz Thompson estimator to yield a population size estimator that is more efficient than the existing ones. Random removals are allowed in the recapturing process. Simulation studies are shown to assess the performance of the proposed estimators. Examples on a bird banding and a small mammal recapturing study are given. PMID- 11892691 TI - Survival analysis using auxiliary variables via multiple imputation, with application to AIDS clinical trial data. AB - We develop an approach, based on multiple imputation, to using auxiliary variables to recover information from censored observations in survival analysis. We apply the approach to data from an AIDS clinical trial comparing ZDV and placebo, in which CD4 count is the time-dependent auxiliary variable. To facilitate imputation, a joint model is developed for the data, which includes a hierarchical change-point model for CD4 counts and a time-dependent proportional hazards model for the time to AIDS. Markov chain Monte Carlo methods are used to multiply impute event times for censored cases. The augmented data are then analyzed and the results combined using standard multiple-imputation techniques. A comparison of our multiple-imputation approach to simply analyzing the observed data indicates that multiple imputation leads to a small change in the estimated effect of ZDV and smaller estimated standard errors. A sensitivity analysis suggests that the qualitative findings are reproducible under a variety of imputation models. A simulation study indicates that improved efficiency over standard analyses and partial corrections for dependent censoring can result. An issue that arises with our approach, however, is whether the analysis of primary interest and the imputation model are compatible. PMID- 11892692 TI - Estimation of anticardiolipin antibodies, anti-beta2 glycoprotein I antibodies and lupus anticoagulant in a prospective longitudinal study of children with juvenile idiopathic arthritis. AB - OBJECTIVE: Anticardiolipin antibodies (aCL) have been frequently detected in juvenile idiopathic arthritis (JIA), but have not been associated with disease activity or clinical features of the antiphospholipid syndrome (APS). Our aim was to determine aCL and anti-beta2 glycoprotein I (anti-beta2GPI) antibody levels and lupus anticoagulant (LA) in serial samples from children with JIA and to investigate the clinical significance of these antibodies. METHODS: The values of aCL, anti-beta2GPI and LA were prospectively followed in 28 children with JIA from disease onset. aCL and anti-beta2GPI were assayed by an ELISA method. Two monoclonal beta2GPI-dependent aCL (HCAL and EY2C9) were used as calibrators. LA was determined by a modified dilute Russell viper venom time test. RESULTS: Thirteen (46.4%) children with JIA were already positive for aCL at their first referral to our center. During the follow-up, the frequency of aCL decreased from 46.4% to 28.6%; however, it remained significantly higher compared with healthy children. In contrast, for anti-beta2GPI the difference in the frequency between the children with JIA and healthy children was not statistically significant. Serial determination of aPL levels in JIA patients revealed frequent fluctuations. Positive aCL persisted over time in 6 (21.4%) children with JIA, 6 (21.4%) children were initially positive for aCL, but became later negative, and 3 (10.7%) children were initially negative for aCL and became later positive. Persistently positive anti-beta2GPI were observed during the follow-up only in one patient, while none of the patients was persistently positive for LA. No association between aCL, anti-beta2GPI or LA and disease activity could be established. No patient with positive aCL, anti-beta2GPI or LA showed any clinical feature of APS. CONCLUSION: The discrepancy between the presence of aCL and anti-beta2GPI might indicate that the production of aCL in JIA is associated with an infectious trigger. Furthermore, the low frequency of anti-beta2GPI and LA could explain the limited prothrombotic potential of aPL observed in JIA. However, we found a distinct group of JIA patients with persistently positive aCL, the clinical implications of which are at the present time unknown. PMID- 11892693 TI - Uveitis as a marker of active arthritis in 372 patients with juvenile idiopathic seronegative oligoarthritis or polyarthritis. AB - OBJECTIVE: To compare the activity of arthritis in children with recently diagnosed seronegative oligoarthritis or polyarthritis with or without uveitis. METHODS: The study covered 372 JIA children with recently diagnosed seronegative oligoarthritis or polyarthritis. The mean prospective follow-up period was 4.5 years. Asymptomatic anterior uveitis was found in 96 cases. The activity of arthritis in all 372 patients was assessed clinically and by laboratory parameters. RESULTS: The erythrocyte sedimentation rate was significantly higher (p = 0.001) at the diagnosis of arthritis and at the end of the follow-up (p = 0.02) in the 96 JIA patients with uveitis than in the 276 JIA patients without uveitis. The hemoglobin value was significantly lower (p = 0.008) at the diagnosis of arthritis in patients with uveitis, but not at the end of the follow up. The number of inflamed joints was significantly greater at the end of the follow-up in patients with persistent polyarthritis and uveitis (p = 0.01) compared to those polyarthritis patients without uveitis. Patients with uveitis were significantly more often treated with oral prednisolone (p < 0.001), glucocorticoid joint injections (p < 0.001), and with methotrexate (p = 0.003) compared to patients without uveitis. Clinical remission of arthritis was achieved significantly less frequently in patients with uveitis than in patients without uveitis (21% versus 42%, p<0.001). CONCLUSION: The inflammatory activity of arthritis seems to be increased in patients with seronegative oligo- or polyarthritis and uveitis compared to those without uveitis. PMID- 11892694 TI - A case of adult onset Still's disease treated with infliximab. PMID- 11892695 TI - Antinuclear antibodies: a marker of susceptibility to autoimmunity? PMID- 11892696 TI - Systemic lupus erythematosus with thyroiditis: a therapeutic dilemma? PMID- 11892697 TI - Etanercept in the treatment of severe, resistant psoriatic arthritis: continued efficacy and changing patterns of use after two years. PMID- 11892699 TI - Antimalarials in the management of psoriatic arthritis? PMID- 11892698 TI - Churg-Strauss syndrome or other disease? PMID- 11892700 TI - Atypical Baker's cyst as a presenting sign of osteomyelitis superimposed on avascular necrosis of the knee. PMID- 11892701 TI - Hepatitis B vaccination and arthritic adverse reactions: a followup analysis of the Vaccine Adverse Events Reporting System (VAERS) database. PMID- 11892702 TI - Anti-nucleosome antibody: significance in lupus patients lacking anti-double stranded DNA antibody. AB - OBJECTIVE: To investigate the clinical significance of anti-nucleosome antibodies in SLE patients lacking anti-double stranded DNA (dsDNA) antibodies. METHODS: IgG anti-nucleosome antibodies were detected by enzyme-linked immunosorbent assays (ELISA) in the sera of SLE patients. Anti-dsDNA antibodies were measured by Farr assays and ELISA, not only in the samples taken for anti-nucleosome testing, but also in sera obtained regularly during the follow-up. RESULTS: Ninety-eight (76.0%) out of 129 patients with SLE had anti-nucleosome antibodies. Twenty-five patients (19.4%) consistently showed little or no anti-dsDNA reactivity during the course of their disease, and among these anti-nucleosome antibodies were present in the sera of 15 (60.0%). Of the patients with anti-dsDNA-negative SLE, renal disorders were present in 8 patients (32.0%), all of whom had anti nucleosome antibodies. Renal disorders were not found in patients (n = 10) who had neither anti-dsDNA nor anti-nucleosome antibodies. Other autoantibodies such as anti-Ro, anti-Sm and anti-cardiolipin were not associated with renal disorders in this group. The levels of anti-nucleosome antibody strongly correlated with the SLEDAI scores, but inversely correlated with serum complement levels in anti dsDNA negative SLE patients. CONCLUSION: Our data suggest that the anti nucleosome antibody may be a useful marker for diagnosis and activity assessment of anti-dsDNA negative SLE. Anti-nucleosome antibody may be an important factor for renal involvement in this subgroup of patients. PMID- 11892703 TI - Comparison of cell-ELISA, flow cytometry and Western blotting for the detection of antiendothelial cell antibodies. AB - OBJECTIVE: There is still great uncertainty in the detection of antiendothelial cell antibodies (AECA). The aim of our study was to compare the results obtained using different methods. METHODS: Sera were obtained from 71 patients with a variety of vasculitides. Three assay methods were used: cell ELISA, flow cytometry (FACS) and Western blot (WB). RESULTS: In the ELISA 12/17 patients with systemic lupus erythematosus (SLE), 1/12 with Churg Strauss (CS) disease, 3/12 with micropolyarteritis (MPA) and 5/30 with Wegener's granulomatosis (WG) tested positive. Most of the sera that were positive on ELISA were not by FACS. Among the negative sera, 50% of WG, 40% of MPA, 20% of CS and 40% of SLE became positive on WB. There were some specific patterns of reactivity for a given disease, so that some bands could be assigned to a disease. CONCLUSION: The discrepancies in the results may most probably be accounted for by differences between the antigenic preparations. Caution must thus be exercised when interpreting the results of any of these three tests. PMID- 11892704 TI - Effectiveness of leukocyte interferon in patients affected by HCV-positive mixed cryoglobulinemia resistant to recombinant alpha-interferon. AB - OBJECTIVE: Interferon is the first-choice therapy for HCV-positive mixed cryoglobulinemia, but only a small fraction of the patients show long-term recovery from the disease. In non-responders or relapsers, the second-line therapy (high dose interferon) generally is not effective. The aim of this study was to evaluate the effectiveness of leukocyte interferon as a second-line therapy in patients who are non-responders or relapsers to a first course of recombinant interferon. METHODS: Twenty-eight patients with HCV-positive mixed cryoglobulinemia were enrolled. In each case the HCV-RNA and HCV genotype, as well as the usual laboratory parameters, were determined before, at the end of therapy and 1 year after the end of therapy. All patients were treated following the same schedule: leukocyte interferon 3,000,000 three times a week for one year. RESULTS: Only 5 patients obtained complete recovery from viral infection as well as from all signs and symptoms of the disease. Most patients (80%) experienced relief from clinical symptoms without recovery from HCV replication. Responders to the second interferon course were "relapsers" to the first treatment. No patient considered as a "non-responder" showed complete remission from the disease after the second treatment. CONCLUSIONS: A second leukocyte interferon course could be useful for patients affected by mixed crvoglobulinemia who relapsed after a first course of recombinant interferon therapy. PMID- 11892705 TI - Rheumatoid arthritis or psoriatic symmetric polyarthritis? A difficult differential diagnosis. PMID- 11892706 TI - Upper gastrointestinal tolerability of celecoxib compared with diclofenac in the treatment of osteoarthritis and rheumatoid arthritis. AB - OBJECTIVE: To compare the upper gastrointestinal (UGI) tolerability of celecoxib (a cyclooxygenase-2 specific inhibitor) and diclofenac using data from three randomised, double-blind clinical trials in osteoarthritis (OA) and rheumatoid arthritis (RA). METHODS: Patients in two OA studies received either celecoxib 100 mg BID (n = 545), diclofenac 50 mg BID or TID (n = 540), or placebo (n = 200) for 6 weeks. In the RA study, patients received celecoxib 200 mg BID (n = 326) or diclofenac 75 mg BID (n = 329) for 24 weeks. The cumulative incidence of abdominal pain, dyspepsia, nausea or any of these events (UGI tolerability composite endpoint) after the first 6 weeks was estimated using time-to-event analysis. RESULTS: In the pooled OA trials, the cumulative incidence of the composite endpoint was significantly higher with diclofenac (17.6%; 95% CI: 14.4 20.9%) than celecoxib (11.1%; 95% CI: 8.4-13.8%; p = 0.002) and comparable with placebo (13.3%; 95% CI: 8.1-18.4%; p = 0.157). In the PA trial, the cumulative incidence of the UGI tolerability composite endpoint was also significantly higher with diclofenac (20.7%; 95% CI: 16.3-25.1%) than celecoxib (15.9%; 95% CI: 11.9-20.0%; p = 0.013). Celecoxib was also better tolerated than diclofenac in this trial in terms of the cumulative incidences of abdominal pain (p = 0.031) and dyspepsia (p = 0.062). The results of the UGI tolerability composite endpoint analysis were confirmed using the Cox proportional hazards model to controlfor other predictors of UGI adverse events. CONCLUSION: The UGI tolerability of therapeutic dosages of celecoxib was significantly better than diclofenac in patients with RA or OA. PMID- 11892707 TI - Autoantibody detection in scleroderma patients. Diagnostic and analytical performances of a new coupled particle light scattering immunoassay. AB - OBJECTIVE: We evaluated the diagnostic and analytical performance of the Coupled Particle Light Scattering technology applied to the detection of anti topoisomerase I (anti-Scl70) and anti-CENP-B autoantibodies. METHODS: The Scl70 antigen was obtained by recombinant DNA procedures using a prokaryotic expression system; CENP-B was a Baculovirus-expressed recombinant protein. Anti-centromere and anti-Scl70 antibodies were assayed in serum samples from 288 patients, of whom 123 had systemic sclerosis/scleroderma and 165 constituted the control groups (including patients with other connective tissue diseases, viral infections, Lyme disease, and healthy subjects). RESULTS: The sensitivity was 98.8% (confidence interval, 96-101%) for anti-Scl70 and 100% (99.6-100%) for anti CENP-B; the specificity was 99.0% (98-100%) and 100% (99.9-100%) for anti-Scl70 and anti-CENP-B, respectively. The intra-assay coefficient variations (CV) ranged from 3.8 to 9.2% for anti-Scl70, and from 2.8 to 8.0% for anti-CENP-B. Inter assay CVs were 8.1 to 12.0% for anti-Scl70, and 4.7 to 10.5% for anti-CENP-B. In 3 patients, coexpression of both antibodies was observed. CONCLUSION: The results of this study demonstrate that the light scattering technology is also applicable to the detection of autoantibodies to intracellular antigens for the diagnosis of autoimmune rheumatic diseases. PMID- 11892709 TI - Efficacy of periarticular corticosteroid treatment of the sacroiliac joint in non spondylarthropathic patients with chronic low back pain in the region of the sacroiliac joint. AB - OBJECTIVE: To investigate the efficacy of periarticular corticosteroid treatment of the sacroiliac joint (SIJ) in non-spondylarthropathic patients with chronic low back pain in the region of the SIJ in a double blind, controlled study. METHODS: Twenty-four consecutive non-spondylarthropathic patients with chronic pain in the region of the SIJ entered the study. Thirteen patients were treated with a periarticular injection of methylprednisoloneacetate and lidocaine (MP group) of the SIJ, whereas 11 patients received isotonic sodium chloride and lidocaine. Clinical assessment at the onset of the study and after one month included the patient's estimation of pain in the region of the SIJ by the visual analogue scale (VAS) and by a pain index, which was calculated from tenderness and stressing tests on the SIJ. RESULTS: At the one month's follow-up examination both the VAS (p = 0.047) and the pain index (0.017) had improved significantly in the MP group compared with the non-MP group. CONCLUSION: These results suggest that periarticular injection of methylprednisolone may be effective in the treatment of pain in the region of the SIJ in non-spondylarthropathic patients. PMID- 11892710 TI - Short-term intravenous therapy with Neridronate in Paget's disease. AB - AIMS: To describe the effects of two consecutive intravenous infusions of aminohexane bisphosphonate (Neridronate) in patients with active Paget's disease of bone. METHODS: The study population included 83 patients, aged 41 to 85 years, randomized to 4 cumulative doses of Neridronate (25, 50, 100, 200 mg) given over 2 days, with a follow up of 180 days. The baseline serum alkaline phosphatase activity was at least 10% above the upper limit of the laboratory range. The response to treatment was assessed by changes in the serum total alkaline phosphatase (primary end point of the study), bone alkaline phosphatase and N telopeptide urinary excretion. RESULTS: All Neridronate doses significantly suppressed the biochemical indices of disease activity. The nadir of total alkaline phosphatase levels ranged from -16 % to -57.5% of pretreatment values in the four groups, with a dose-response relationship that was apparent even between the two highest doses. The proportion of patients still maintaining a partial response (decreases in serum total alkaline phosphatase >25%) at the 6 month follow-up was also related to the dose: 98%, 67%, 57%, 21% in the patients given 200, 100, 50, 25 mg respectively. The proportion of responders in terms of bone alkaline phosphatase and N-telopeptide excretion changes was similar. Bone pain attributed to Paget's disease was significantly reduced. A typical acute phase reaction (fever and/or arthromyalgia) occurred in 16 out of 83 patients. CONCLUSIONS: We conclude that all of the Neridronate doses tested here were well tolerated and effective in decreasing, in a dose-related manner the bone turnover parameters of Paget's disease. The highest dose (200 mg) resulted in the normalization of the markers of disease activity in more than 60% of the patients. PMID- 11892711 TI - The role of peroxynitrite in cyclooxygenase-2 expression of rheumatoid synovium. AB - OBJECTIVE: Reactive oxygen intermediates play an important role in the inflammatory processes of rheumatoid arthritis. Cyclooxygenase-2 is an inducible form of an enzyme involved in prostanoid biosynthesis. This study linked peroxynitrite (ONOO-) to the signaling pathways that induce COX-2. RESULTS: Exposure of rheumatoid synovial cells to peroxynitrite resulted in COX-2 protein expression in a dose-dependent manner. RT-PCR analysis also demonstrated that COX 2 mRNA was induced in peroxynitrite-treated rheumatoid synovial cells. Dexamethasone markedly inhibited this peroxynitrite-mediated COX-2 expression at therapeutic concentrations. CONCLUSION: This study demonstrates that oxidant stress is an important inducer of COX-2 in rheumatoid synovium. This induction may contribute to the amplification of prostanoids in the rheumatoid inflammatory process. PMID- 11892712 TI - Rheumatoid arthritis and B-cell chronic lymphocytic leukemia. AB - The association between lymphoproliferate malignancies, especially lymphoma, and rheumatoid arthritis (RA) has been confirmed by several studies. However; there are few reports of RA patients who developed B-cell chronic lymphocytic leukemia (B-CLL) and vice versa. We report a patient with B-CLL who developed RA and another with RA who presented with B-CLL during follow-up. We discuss the incidence of B-CLL among the RA population and the possible interaction of the pathogenetic mechanisms of these two entities. PMID- 11892713 TI - Traumatic spinal cord injury as a complication to ankylosing spondylitis. An extended report. AB - OBJECTIVE: Kapyla Rehabilitation Centre is in Finland the only unit taking care of the subacute rehabilitation activities of patients with spinal cord injury (SCI). The annual incidence of new patients with SCI is 55 (1.1 per 100,000 inhabitants). The ankylosed spine (AS) is reported to be at greater risk for fracture and SCI. The aim of the study was to clarify if this higher risk of ankylosing spondylitis (AS) could also be detected among patients with traumatic SCI rehabilitated at Kapyla Rehabilitation Centre. Further, the aim was to evaluate the characteristics of patients with traumatic SCI as a complication to AS in order to develop prevention of SCI in patients with AS. METHODS: Patient data was gathered from the patient register covering all Finnish patients with traumatic SCI (n = 1,103) rehabilitated at Kapyla Rehabilitation Centre from the year 1979 to 1998. The patient journals were subjected to a detailed and systematic analysis. Data about patients with a history of AS (n = 19; 18 men, 1 woman) was then compared to the data about all the patients with SCI (n = 1,103; 902 men, 201 women). RESULTS: Based on the national prevalence data, the incidence rate of patients with AS for traumatic SCI was found to be 11.4 times greater than expected for the population at large. The mean age of the patients with AS was clearly higher (55.3 yrs) than the mean age of the whole group of patients (36.4 yrs) with traumatic SCI. The neurologic injury was at the cervical level in 84% of the patients with AS, but only in 48% of the patients with traumatic SCI in general. Among the patients with AS, the SCI was caused by slipping in 53% of the cases, whereas slipping was the reason for SCI only in 7% of the cases in general. CONCLUSION: Patients with AS seem to run a higher risk of traumatic SCI than the people at large, and the injury levels are higher. In particular, male patients with advanced AS should be instructed to install preventive devices such as night lights and handrails, supports or head rests when driving a car, and they should avoid walking on slippery surfaces, loose carpets etc. They also should be encouraged to avoid excessive use of alcohol and activities involving the risk of physical injury such as contact sports. PMID- 11892714 TI - Hypersomnolence in fibromyalgia syndrome. AB - OBJECTIVE: To evaluate hypersomnolence in patients affected by fibromyalgia syndrome. METHODS: Thirty consecutive patients affected by fibromyalgia syndrome (FMS) (28 F) completed a sleep questionnaire and underwent the following evaluations: lung function tests; polysomnography; the Epworth sleepiness scale (ESS), which measures sleep complaints and daytime hypersomnolence; and the visual analogical scale (VAS) to detect subjective pain, fatigue, anxiety and depression. RESULTS: The FMS patients were divided into two groups based on their ESS score. Patients complaining of daytime hypersomnolence had a higher number of tender points (15 +/- 2 vs. 12 +/- 1, p < 0.01), a higher subjective pain score (72 +/- 15 vs. 52 +/- 13, p < 0.05), and more fatigue (p < 0.05). The diffusing capacity of the lung (Tlco) was more impaired and the occurrence of periodic breathing was higher. FMS patients complaining of daytime somnolence had significantly less efficient sleep than the FMS patients with no daytime somnolence (p < 0.05), i.e. a lower proportion of stage 3 sleep (5 +/- 2% vs. 12 +/- 3%; p < 0.001), stage 4 sleep (1 +/- 0.5% vs. 4 +/- 1%; p < 0.001), and twice as many arousals per hour of sleep (p < 0.01). The respiratory pattern of FMS patients with hypersomnolence showed a higher occurrence of periodic breathing (p < 0.05). The short length of apneas and hypopnoeas did not affect the apnea/hypopnea index (5.1 +/- 3 vs. 7 +/- 4; ns), but FMS patients with daytime hypersomnolence had a greater number of desaturations per hour of sleep (11 +/- 6 vs. 6 +/- 5; p < 0.05). Pulmonary volumes did not differ between the two groups. The EES score was significantly correlated in FMS patients, and even more markedly in the FMS patients with hypersomnolence, TLco, A/I, and disease duration. The ESS score was correlated significantly with the number of tender points only in FMS patients with daytime hypersomnolence. CONCLUSION: The occurrence of daytime hypersomnolence in FMS patients is linked to a greater severity of fibromyalgia symptoms and to more severe polysomnographic alterations. PMID- 11892715 TI - Intralesional therapy in carpal tunnel syndrome: a sonographic-guided approach. AB - OBJECTIVE: The aim of this pictorial essay is to show a representative example of sonographic-guided injection in carpal tunnel syndrome associated with tenosynovitis of the finger flexor tendons. METHODS: Images were obtained using a real-time ultrasound system (AU4-idea; Esaote Biomedica, Genoa, Italy) equipped with a 13-MHz linear transducer. The best injection site was detected using a fine metal clip placed between the skin and the transducer. The images here were obtained in a patient with rheumatoid arthritis and carpal tunnel syndrome secondary to tenosynovitis of the finger flexor tendons. RESULTS: Steroid injection within the carpal tunnel under sonographic control was easily performed. All steps of the needle placement within the widened tendon sheath were carefully evaluated on the monitor screen. Marked clinical improvement occurred shortly thereafter (3 days) and increased over the next 6 weeks. CONCLUSION: A detailed assessment of the carpal tunnel and a correct, safe placement of the needle for steroid injection can be quickly performed under sonographic guidance. PMID- 11892716 TI - Co-occurrence of spondyloarthropathy and connective tissue disease: development of Sjogren's syndrome and mixed connective tissue disease (MCTD) in a patient with ankylosing spondylitis. AB - Spondylarthropathies (SpA) and connective tissue diseases (CTD) are clinically distinct entities which, at first glance, seem to have little in common. However, a link between SpA and CTD has recently been suggested by a study in which a higher prevalence of Sjogren's syndrome (SS) and sicca symptoms was reported in patients with ankylosing spondylitis (AS) and undifferentiated SpA (1). Another link between SpA and CTD is a possible side effect of a DMARD widely used to treat SpA: sulfasalazine (SAS). SAS was reported to induce antinuclear antibodies (ANA) and systemic lupus erythematosus (SLE)-like syndromes such as drug-induced lupus. This report describes a 54-year-old white male, HLA B27-positive AS patient with some syndesmophytes who, after 15 years of disease, developed SS with salivary gland involvement, Raynaud's syndrome and anti-Ro antibodies. Then, 20 years after the onset of AS, he became acutely ill, suffering severe myositis and myocarditis along with swollen hands and highly elevated autoantibody titers recognizing UIRNP; his condition was interpreted as mixed connective tissue disease (MCTD). The patient had been treated with SAS and azathioprine (AZA) alone several times during the last years because he had not tolerated other DMARDs. A combination of both drugs had been prescribed 3 weeks before a severe flair because of progredient high disease activity with painful peripheral arthritis of the MCP and PIP joints which, however, had not shown radiographic erosions. We describe the rare development of MCTD in an AS patient and report, for the first time, the onset of MCTD potentially triggered by sulfasalazine. PMID- 11892717 TI - Marked and sustained improvement of systemic sclerosis following polychemotherapy for coexistent multiple myeloma. AB - We present the case of a patient who had systemic sclerosis (SSc) with progressive cutaneous and pulmonary involvement and a coexisting monoclonal gammopathy which gradually progressed to overt multiple myeloma. Strikingly, successful VMCP (vincristine, melphalan, cyclophosphamide and prednisolone) polychemotherapy for myeloma was accompanied by a marked and sustained improvement of SSc that has persisted for 2 years after chemotherapy. Most noticeable was a substantial skin softening in our patient, with a > 50% reduction in the skin thickness score following VMCP treatment. Our case suggests that polychemotherapy may represent a promising treatment option in patients with SSc who are refractory to available treatments. PMID- 11892718 TI - Vasculitis with mesangial IgA deposits complicating relapsing polychondritis. AB - The authors report the case of a patient presenting with cutaneous, renal and neurologic vasculitis in the course of relapsing polychondritis (RPC). A 78-year old man presented with a palpable purpura of the lower limbs, high fever arthralgias, delirium, and nephrotic syndrome. He had a history of relapsing polychondritis treated by corticosteroids. Renal biopsy showed diffuse endo- and extracapillary proliferative glomerulonephritis with mesangial IgA deposits. A spectacular regression of the symptoms was observed in response to pulse intravenous methylprednisolone. Relapsing polychondritis is complicated by vasculitis in 25% of the cases. This vasculitis is characterized by cutaneous, neurologic and renal manifestations, usually occurring in elderly patients. Renal involvement is characterized by segmental and focal or diffuse necrotizing glomerulonephritis. The mesangial IgA deposits observed in our patient are rarely present in the course of RPC. Renal manifestations identify severe forms of RPC, justifying systematic screening for renal complications. PMID- 11892719 TI - Knee osteoarthritis: the influence of environmental factors. AB - Osteoarthritis is a major source of disability in developed countries. As populations age, we can expect knee OA to become a serious public health problem. Preventative strategies to minimise the risk of both the development and progression of knee OA are therefore of paramount importance not only with respect to quality of life issues but the burdening costs of managing and treating this common disorder in the next few decades. The focus of preventative strategies should be on the modifiable risk factors for both incident and progressive disease. The aetiology of OA of the knee is complex and multifactorial. This review focuses on the modifiable environmental risk factors for knee osteoarthritis; namely occupation, physical activity, quadriceps strength, joint injury, obesity, diet, sex hormones, and bone density. Their contribution is well understood and the impact of altering their influence on knee OA outcome is also now being evaluated. Since they are modifiable, this has important implications for public health recommendations and treatment by health professionals. PMID- 11892720 TI - Heart rate variability as predictor of nonresponse to mirtazapine in panic disorder: a preliminary study. AB - Using spectral analysis of heart rate, several studies have shown that panic disorder patients are characterized by a reduced heart rate variability (HRV), indicative of abnormalities in autonomous nervous system (ANS) function. We recently reported that patients with panic disorder, who did not respond to pharmacotherapy, were characterized at baseline by a higher heart rate. In this study, ANS functioning is investigated as a possible predictor of nonresponse to pharmacotherapy. Twenty-eight medication-free panic disorder patients entered a 12-week open-label treatment study with mirtazapine. Five-minute HRV recordings were obtained before treatment and were analysed using spectral analysis. The data of 17 patients could be used. The total spectrum and low frequency power of responders to mirtazapine were significantly higher than those of nonresponders. Our findings suggest that nonresponders to short-term mirtazapine treatment are characterized at baseline by a lowered output of the ANS. The results are preliminary in view of the small sample studied. PMID- 11892721 TI - Extrapyramidal syndromes associated with selective serotonin reuptake inhibitors: a case-control study using spontaneous reports. AB - The aim of this study was to assess whether use of selective serotonin reuptake inhibitors (SSRIs) is associated with extrapyramidal syndromes (EPS). We analysed the spontaneous reports of adverse drug reactions (ADRs) collected by The Netherlands Pharmacovigilance Foundation Lareb in the period 1985-99 (n = 24,263). The study population comprised all patients using an antidepressant drug at the time the ADR occurred. We calculated ADR-reporting odds ratios (ADR-OR) to estimate the association between SSRI-use and EPS, relative to other antidepressants. We identified 61 patients with EPS. SSRI-use was associated with spontaneous reporting of EPS compared to other antidepressants (adjusted ADR-OR 2.2; 95% confidence interval 1.2-3.9). This risk estimate appeared to be higher in patients concurrently using antipsychotic medication (6.9, 0.7-68.0), although the confidence interval was very wide. In conclusion, SSRI-use seems only to be moderately associated with EPS compared to other antidepressants. However, those concurrently using antipsychotic drugs or presenting with other risk factors may be more susceptible and should be closely monitored. PMID- 11892723 TI - Deficient dietary vitamin K intake among elderly nursing home residents in Hong Kong. AB - There is strong evidence supporting the importance of vitamin K in bone health and the aetiological role of vitamin K deficiency in osteoporosis. In view of the common occurrence of osteoporosis among older subjects in Hong Kong, we have studied the dietary vitamin K intakes in 100 residents of a nursing home (43 men, 57 women; median age 81.0 years) and 88 free-living subjects attending a day care centre (13 men, 75 women; median age 71.5 years). The subjects were interviewed and the average vitamin K intake in the preceding week was estimated, using a diet recall questionnaire modified from our previous surveys of dietary patterns in local Chinese people. The median vitamin K intake was much lower in nursing home residents than in free-living subjects (4.50 vs 488.09 microg/day or 0.13 vs 8.74 microg/kg/day, P<0.001). An intake that was below the recommended daily intake was far more common among nursing home residents (86.0 vs 11.4%, P < 0.001). Among nursing home residents, there was a negative correlation between age and vitamin K intake (r = -0.217, P = 0.030), but there was a positive correlation between body weight and vitamin K intake (r = 0.244, P = 0.015). No such relationship was seen among free-living subjects. Elderly nursing home residents in this study generally had a poor dietary vitamin K intake and might therefore be predisposed to osteoporosis. The importance of green leafy vegetables as a rich source of vitamin K should be emphasised. PMID- 11892722 TI - A valid food frequency questionnaire for measuring dietary fish intake. AB - There is considerable interest in the potentially protective effects of high fish consumption on many chronic diseases. Many epidemiological studies use food frequency questionnaires (FFQ) to quantify usual dietary fish intake, so it is important to validate this assessment against objective markers. The objective of this study was to determine the relationship between plasma percentage fatty acids and dietary fish intake as assessed by a FFQ. A semiquantitative FFQ was completed by 174 adults from the community (aged 26-49 years) who also had venous blood analysed for plasma percentage fatty acids. Following linear regression modelling, total non-fried fish intake was a significant predictor of n-3 (regression coefficient, B = 0.94; 95% CI = 0.60-1.28), docosahexaenoic acid (DHA; B = 0.73; 95% CI = 0.47-0.99) and the ratio of n-6: n-3 fatty acids (B = 1.0; 95% CI = - 1.35- -0.65). Steamed, grilled or baked fish was a small but significant predictor of eicosapentaenoic acid (EPA) levels (B = 0.13; 95% CI = 0.05-0.21) while total fish intake was a predictor of n-6 fatty acids (B = -0.88; 95% CI = -1.41- -0.36). This semiquantitative FFQ could be useful for ranking subjects according to their likely plasma n-3, DHA, and n-6 fatty acid intake and the ratio of n-6: n-3 fatty acids, when the available resources may simply not permit biological markers to be used. PMID- 11892724 TI - Comparison of latex agglutination with enzyme immunoassay for detection of rotavirus in fecal specimens. AB - Human rotaviruses are the most important etiologic agents of acquired diarrhea in infants and young children worldwide. Early diagnosis is essentialfor effective patient treatment. The latex agglutination (LA) assays for rotavirus diagnosis are rapid, inexpensive, and the most widely used to screen specimens. The performance of the LA Rotagen (Biokit S.A., Barcelona, Spain) was evaluated for rotavirus detection infecal samples of outpatients with acute gastroenteritis. This assay was compared with the enzyme immunoassay (EIA) EIARA (Bio-Manguinhos, Rio de Janeiro, Brazil). From January to October 2000, 285 fecal specimens were analyzed. Forty-four samples (15.4%) were reactive, 214 (75.4%) were nonreactive, and 27 (9.5%) were indeterminate by LA. All LA-positive samples were positive by EIA, and 2 LA-negative samples were positive by EIA. Of specimens indeterminate by LA, 67% were positive by EIA. The sensitivity, specificity, and accuracy of LA were 69%, 100%, and 93%, respectively. These results indicate that assay is as sensitive and specific as the EIA, and it could be applied on a large scale for screening stool specimens in suspected rotavirus diarrhea. However, the indeterminate results must be confirmed by other methods, such as EIA. PMID- 11892725 TI - Clinicopathologic and prognostic significance of the histologic activity of noncancerous liver tissue in hepatitis B virus-associated hepatocellular carcinoma. AB - We prospectively studied 66 patients infected with the hepatitis B virus who underwent liver resection for hepatocellular carcinoma (HCC) to evaluate the influence of the histologic activity of noncancerous liver tissue on clinicopathologic features and prognosis. Based on the histologic activity index (HAI) score of nontumorous liver tissue, patients were classified into 3 groups: mild, moderate, or severe hepatitis. Overall, higher HAI scores were more frequent in patients with poorer liver function: lower serum albumin levels and higher indocyanine green retention at 15 minutes. Moreover, patients with moderate hepatitis had more frequent venous invasion, and the tumor size decreased with increasing HAI scores. Similar results were observed when the fibrosis category was excluded in the calculation of HAI scores. The overall or disease-free survival rates did not differ significantly among the 3 groups of patients. However, higher fibrosis scores were associated significantly with shorter disease-free survival rates. HAI scores correlated significantly with certain clinicopathologic features. In patients with hepatitis B-related HCC, a higher fibrosis score in the nontumorous liver tissue, but not histologic hepatitic activity, seems to be a significant factor predisposing to shorter survival. PMID- 11892726 TI - Safety assessment and public concern for genetically modified food products: the European view. AB - The safety assessment for marketing purposes of genetically modified (GM) foods in the 15 Member States of the European Union (EU) is based on the Novel Foods and Novel Food Ingredients Regulation adopted in May 1997. Before a GM food can be approved under the Regulation, it must satisfy three criteria: Gm food must be safe, it must not mislead the consumer and it must be nutritionally adequate. The EU Scientific Committee on Food has published a set of guidelines describing the type of information expected from a company in support of an application for approval of a GM food or food ingredient. Despite this rigorous procedure and there being no evidence of harm resulting from the consumption of GM foods worldwide, there is essentially no market in the EU for such products at present. Possible reasons for this are discussed and the view put forward that the market for GM foods will change only when there are more clearly perceived consumer benefits. PMID- 11892727 TI - Endocrine active agents: implications of adverse and non-adverse changes. AB - The US Environmental Protection Agency (EPA) is currently in the process of developing screening and testing methodologies for the assessment of agents that may possess endocrine-like activity--the so-called endocrine disruptors. Moreover, the EPA has signaled its intention of placing information arising from such studies on the worldwide web. This has created significant interest in how such information may be used in risk assessment and by policymakers and the public in the potential regulation or deselection of specific chemical agents. The construction of lists of endocrine disruptors, although fulfilling the requirements of some parties, is really of little use when the nature of the response, the dose level employed, and the lifestage of the test species used are not given. Thus, we have already seen positive in vitro information available on the interaction with a receptor being used as a key indicator when the results of large, high quality in vivo studies showing no adverse changes have been ignored. Clearly a number of in vitro systems are available to ascertain chemical interaction with specific (mainly steroid) hormone receptors including a number of reporter gene assays. These assays only provide indicators of potential problems and should not be, in isolation, indicators of toxicity. Likewise, short term in vivo screens such as the uterotrophic and Hershberger studies are frequently conducted in castrated animals and thus indicate the potential for a pharmacological response in vivo rather than an adverse effect. A number of new end points have been added to standard rodent testing protocols in the belief of providing more sensitivity to detect endocrine related changes. These include the measurement of anogenital distance (AGD), developmental landmarks [vaginal opening (VO), preputial separation (PPS)], and in some studies the counting of nipples and areolae on males. AGD, VO, and PPS are all affected by the size of the pup in which they are measured and should always be compared using bodyweight as a covariate. The historical control database for such changes is gradually growing, albeit that if pups are not individually identified it becomes problematic to associate any change with a specific malformation or to assess whether a delay or advance in, for example, developmental landmarks is biologically significant. Agents that significantly reduce AGD in males (it is an androgen-dependent variable) frequently have other more adverse changes associated with this end point (eg, reproductive tract malformations), but a 2 to 3% change in AGD although measurable is unlikely to be biologically of importance and in isolation would not necessarily be considered adverse. Retention of thoracic nipples in male rat pups is also an indicator of impaired androgen status. Recent studies have also shown that this retention for some endocrine active chemicals is permanent. Thus, the presence of a permanent structural change that is rarely found in adult control animals could be considered a malformation and therefore a developmental adverse effect on which risk assessment decisions could be made. The advent of multigeneration reproduction studies as the definitive studies for the assessment of the dose-response relationships and risk assessment for endocrine disruptors has shown that current testing protocols may be inadequate to reliably detect the adverse effects of concern as only 1 adult/sex/litter is examined. A number of the effects on reproductive development although, due to an in utero exposure, will not be manifest until after puberty or at adulthood. The use of only a limited number of animals to examine such changes, particularly for weaker acting materials indicates that some agents may have been examined in well-conducted, modern protocols but have insufficient power to detect low incidence phenomena (eg, a 5% incidence of malformations). PMID- 11892728 TI - Consequences of prolonged wait before gallbladder surgery. AB - The aim of this study was to document the morbidity associated with long waiting times for laparoscopic cholecystectomy and to relate this to the nature of initial presentation either routine out-patient consultation or emergency admission with acute symptoms. This study was performed over a 50-month period in a DGH (serving a population of 320,000) which lacked sufficient operating capacity to allow routine early cholecystectomy after emergency admission. A total of 387 patients underwent cholecystectomy but 22 of these had an early operation after initial emergency admission with signs of peritonitis and were excluded from the study. The median waiting time for cholecystectomy in this study population of 365 patients was 170 days (range, 6-484) days. Of these 365 patients, 246 (67.4%) were listed for surgery after initial out-patient assessment (out-patient cohort) and 119 (32.6%) were diagnosed after an index emergency admission with symptomatic gallstone disease (emergency cohort). Of the 365 patients, 42 (11.5%) had one or more emergency admissions (57 admissions) with gallstone-related complications whilst on the waiting list for surgery. Complications were acute cholecystitis/biliary colic (n = 40), jaundice/cholangitis (n = 8), acute pancreatitis (n = 6) and perforated gallbladder (n = 3). Re-admissions with gallstone-related complications were much more common in patients whose initial presentation had been as an emergency. Thus, 34 of the 119 emergency cohort (28.5%) required re-admission with complications whilst only 8 of 246 (2.8%) elective cohort were re-admitted. Of the 34 re-admissions in the emergency cohort, 22 occurred within 6 weeks of their discharge from hospital. Median total and postoperative stay were significantly shorter (P < 0.001) in the elective cohort (3 and 2 days, respectively) than the emergency cohort (10 and 3 days, respectively). These results document the high incidence of complications experienced by patients waiting for an elective laparoscopic cholecystectomy. Morbidity is highest in patients with an initial emergency admission. These results suggest that cholecystectomy should be offered to all patients presenting as an emergency with symptomatic gallstones on admission. PMID- 11892729 TI - Dukes' staging is poorly understood by doctors managing colorectal cancer. AB - OBJECTIVE: This study set out to investigate the current understanding of Dukes' staging for colorectal cancer. DESIGN: A questionnaire was distributed to surgeons and general practitioners attending colorectal meetings asking for a definition of Dukes' stages A, B and C. Results were analysed blind by two authors jointly to assess accuracy as correct, within definition, or incorrect. Within definition was defined as a description fitting within but not covering all tumours within that stage. RESULTS: 128 answers were received from 48 GPs, 7 final year medical students, 38 house officers and SHOs, 19 higher surgical trainees and 16 consultants. Overall, 3.9% defined all three stages correctly and 13.3% got all three definitions incorrect. Correct stages were Dukes' A 7.8%, Dukes' B 16.4% and Dukes' C 29.7%. Two consultants (12.5%) achieved three correct definitions, as did two HSTs (10.5%). No GPs had all three stages correct and 10 (20.8%) were wrong in all three. If those said to be within definition were considered right, 35.1% were correct for all three stages with 76.6% getting Dukes' A correct, 46.9% Dukes' B and 56.6% Dukes' C. CONCLUSIONS: Dukes' staging is, therefore, still poorly understood by doctors managing patients with colorectal cancer. The introduction of proformas will reduce the reliability upon memory for this and more complex staging classifications. PMID- 11892730 TI - Complications following peripheral angioplasty. AB - BACKGROUND: Peripheral angioplasty is increasingly the first choice intervention in patients with peripheral vascular disease. The aim of the current study was to audit prospectively all major complications, especially the requirement for emergency surgical intervention. PATIENTS AND METHODS: A prospective audit of outcome after peripheral angioplasty in 988 patients undergoing 1377 interventional procedures between 1 October 1995 and 30 September 1998 at which 1619 vessel segments were angioplastied. RESULTS: Major medical morbidity (bronchopneumonia, stroke, renal failure, myocardial infarction) complicated 33/1377 procedures (2.4%). Emergency surgical intervention was required after 31/1377 procedures (2.3%) with the commonest aetiologies being acute limb ischaemia and haemorrhagic complications. The amputation rate following angioplasty was 0.6% and no patient presenting with claudication or graft complications underwent amputation. The amputation rate following angioplasty for critical limb ischaemia was 2.2%. Overall, the risk of death and/or major medical complication and/or requiring emergency surgical intervention was 3.5%. The rate of complications was no different for subintimal as opposed to transluminal angioplasties. CONCLUSIONS: Peripheral angioplasty is associated with a low risk of major medical and surgical complications. PMID- 11892731 TI - Can a district general hospital serving a population of 480,000 offer subspecialty training? --A prospective audit. AB - BACKGROUND: Subspecialty training has been mostly restricted to teaching hospitals. We aimed to assess whether higher surgical trainees can be offered subspecialty training in a district general hospital serving a large population. METHODS: The surgical unit consisted of four subspecialty firms (upper gastrointestinal, vascular, colorectal and breast/endocrine). Each firm consisted of two consultants, one higher surgical trainee and one basic surgical trainee. The breast/endocrine firm had, in addition, a staff grade surgeon. Trainees collected data prospectively on their subspecialty experience and this was then compared with the subspecialty workload in the respective firms. RESULTS: Subspecialty related workload was 48% on the vascular, 57% on the colorectal and 53% breast/endocrine firms. Subspecialty workload on the upper gastrointestinal firm (27%) was skewed by one non-specialist consultant Trainees on the respective firms were involved in 74% vascular, 82% upper gastrointestinal, 79% colorectal and 54% breast/endocrine index subspecialty operations. Supervision with regards to index operations was 63%, 70%, 81% and 100% on the colorectal, breast/endocrine, upper gastrointestinal and vascular firms, respectively. CONCLUSIONS: 50% of the workload on the vascular, breast/endocrine and colorectal firms is subspecialty-related with the potential for training. With shortened training and some specialities having disproportionately more trainees, higher surgical training committees need to identify more subspecialty units that offer such training. PMID- 11892732 TI - Do we need to improve awareness about HIV post exposure prophylaxis? PMID- 11892733 TI - [Effect of head-down bed rest on human EEG]. AB - Objective. To study the influences of short-term simulated weightlessness on human EEG. Method. EEG of 12 male subjects (aged 18-22) was recorded, pre-HDBR ( 6 degrees head down bed rest), mid-HDBR (3rd, 4th, 6th day) and post-HDBR (3rd day). Then these EEG recordings were analyzed by power spectral analysis. Result. During HDBR, peak frequency of EEG gradually slowed down (6th day, P<0.05). Alpha band activities were enhanced (alpha1: P<0.001. Alpha2: 3rd and 6th day, P<0.05; 4th day, P<0.001) and reached maximum on the 4th day. Alpha1/alpha2 ascended (6th day, C3 and C4: p<0.02; F4, P4, T3, T4, T5: P<0.05). Theta-band activities enhanced (P<0.001), and reached maximum on the 3rd day at frontal regions, on the 4th day at parieto-occipital regions. Theta/alpha ascended obviously (6th day, P3, O1: P<0.05). Meanwhile, beta1, activities also enhanced (P<0.001), and reached maximum on the 3rd day at frontal regions. After HDBR, the index that recovered to the pre-HDBR level first was EEG peak frequency. Conclusion. HDBR caused obvious changes of EEG power spectrum characteristic. There exists a potential influence on brain function. PMID- 11892734 TI - [Transition of soleus troponin I isoforms and atrophy of testis in tail-suspended rats]. AB - Objective. To observe the transition time of soleus I (TnI) isoforms and to elucidate the relationship between soleus TnI transition and atrophy; and to analyze the time course between testis atrophy and soleus atrophy. Method. Eight groups of male rats were suspended for 3, 4, 5, 7, 14, 21, 28 and 42 d, respectively. Besides, three groups of female rats were suspended for 3, 4 and 5 d respectively. Wet and relative weights (wet weight/body weight) of testis and soleus were measured. The expression of TnI was observed by Western blot. Result. The relative weight of soleus of tail-suspended male rats decreased significantly after 4 d of suspension as compared with control. The degree of rats soleus atrophy in the first 14 d of suspension was greater than that after 14 d. The relative weight of testis showed the same change as that of soleus. There was no significant change in the relative weight of soleus in 4 d of tail-suspended female rats. The significant decrease in the relative weight of tail-suspended female rats began on the 5th day. The Western blot showed that the transition from slow skeletal TnI (ssTnI) to fast skeletal TnI (fsTnI) in the soleus occurred in 14 d of tail-suspension. Conclusion. The overt atrophy of tail suspended male rats occurs on the 4th day. The soleus TnI transition from ssTnI to fsTnI is on the 14th day. It is suggested that the TnI is not the sensitive protein to gravity. The overt atrophy of female tail-suspended rats occurs at the 5th day. This indicated that the decrease in testosterone level may accelerate the atrophy of the soleus. PMID- 11892735 TI - [Estimating cardiovascular age of civil flying personnel by means of heart rate and blood pressure variability analysis]. AB - Objective. To estimate the cardiovascular age of civil flying personnel by means of heart rate and blood pressure variability analysis and to evaluate its significance in aviation medicine. Method. First, heart rate variability (HRV), blood pressure variability (BPV) and spontaneous baroreflex sensitivity (BRS) were analyzed among 89 healthy civil flying personnel by using conventional AR spectral analysis and sequence method respectively. Then, principal component analysis was conducted over original and derived variables of HRV and BPV spectral and BRS data. Finally, by the use of multiple regression in which the chronological age acted as the dependent variable and the components significantly related to age were used as the regressors, the equation for estimating the cardiovascular age was established. Result. Only seven principal components can exactly reflect the same information of autonomic regulatory function which was embodied in the 17 variables of HRV and BPV spectral and BRS parameters. Among the seven principal components, the PC2orig, PC4orig and PC2deri were negatively correlated with chronological age (P<0.05), whereas the PC3orig was positively correlated with the chronological age (P<0.01). The cardiovascular age derived from the equation was significantly correlated with the chronological age of the civil flying personnel (r= 0.73, P<0.01). Conclusion. The cardiovascular age estimated by means of a multi-variate analysis of HRV, BPV and BRS can be treated as a comprehensive indicator reflecting the age dependency of autonomic regulatory function of cardiovascular system in healthy civil flying personnel, and its interpretation and significance in application are surely worthy of further and fully dedicated efforts. PMID- 11892736 TI - [Application of degree of complexity in heart rate variability analysis during orthostatic (correction of orthosatic) standing]. AB - Objective. To introduce the degree of complexity in characterizing heart rate variability (HRV), and to discuss the changes in complexity of the cardiovascular system during orthostatic standing posture. Method. ECG of 8 subjects were recorded during supine and orthostatic standing postures. Degree of complexity was used to analyze the HRV. Result. Compared to supine before orthostatic standing posture, R-R intervals and its standard deviation at 0-5 min, 5-10 min, 10-15 min and 15-20 min during orthostatic standing posture were decreased significantly; the degree of complexity of HRV at 0-5 min and 15-20 min during orthostatic standing posture were decreased significantly; the approximate entropy of 0-5 min and 15-20 min were decreased significantly. Conclusion. The results showed that HRV and complexity of cardiovascular system was decreased during orthostatic standing. It was feasible that the degree of complexity can be used to analyze HRV. PMID- 11892737 TI - [Principle demonstration of nutrient delivery system in a space vegetable planting prototype facility]. AB - Objective. To develop a nutrient delivery system for space vegetable planting prototype facility to be used in future space station, and to preliminarily testify its feasibility through ground-based demonstration experiments. Method. A nutrient delivery system in a space vegetable planting prototype facility was designed and fabricated, and ground based demonstration experiments of plant cultivation were conducted. Result. Nutrient could be steadily delivered to plant cultivation matrixes through capillary action, water content of planting matrixes could be controlled automatically and maintained constant, and the planted material lettuce showed basically normal morphology and color. Conclusion. The nutrient delivery system in a space vegetable planting prototype facility could basically meet the requirements for plant nutrient delivery under space microgravity environmental condition. PMID- 11892739 TI - [Locating the impairment of human cognitive function during hypoxia]. AB - Objective. To find the location where the human brain cognitive function impairs under hypoxia. Method. 14 healthy males, aged 18-20 years, performed auditory Oddball test of two different intensities (55 dB, 80 dB) during exposure to 5000 m by breathing low oxygen mixture. EEG and Reaction Time (RT) were recorded. P3, the component of ERP, was extracted from EEG. P3 latency and RT were used as indices of the experiment. Additive Factors Method was used to analyse the result of the experiment. Result. Interaction between hypoxia and stimulus intensity was found for P3 latency and RT. Conclusion. Acute hypoxia influences the preprocessing stage of information processing. PMID- 11892738 TI - [Protective effects of Chinese herb-compound on cellular immunological function (correction of funcion) in tail-suspended rats]. AB - Objective. To observe the protective effects of two kinds of Chinese herb compounds (Dan-huang-ci compound and Shen-chuan-shu compound) on cellular immunology in tail-suspended rats. Method. The rats were divided into: 1) normal control group; 2) tail-suspended group; 3) tail-suspended + Dan-huang-ci compound; and 4) tail-suspended + Shen-chuan-shu compound. Ability of lymphocyte proliferation and production of IL-2 in rats in the four groups were compared after 21 d. Result. The immunological function of tail-suspended control group decreased significantly as compared with normal control group. Shen-chuan-shu compound could improve immunological function of tail-suspended rats obviously. Conclusion. Shen-chuan-shu compound could enhance cellular immunological function in rats under simulated weightlessness. PMID- 11892740 TI - [Effects of adaptive changes of vestibular system on cardiovascular regulation and orthostatic tolerance]. AB - Astronauts may experience Space Adaptation Syndrome at the primary period in spaceflight. It is widely accepted that it is mainly caused by vestibular adaptive changes. The reverse changes of adaptation present after the return from weightlessness to normal 1 G. It may cause astronauts' postural instability and orthostatic intolerance. Evidences from animal model and some data from humans suggest that the vestibular system have influences on the sympathetic nervous system and cardiovascular control. The inputs that appear to be critical in producing these responses come from otolith receptors. The important medical problem--orthostatic intolerance in space medicine is mainly caused by cardiovascular abnormal control. It is possible that vestibular adaptive changes under microgravity may have its influence on the cardiovascular function and orthostatic intolerance. PMID- 11892741 TI - [Research progress of methods of heart rate variability analysis]. AB - One of the hotspots in the analysis of ECG is heart rate variability (HRV). In the present, the method mostly used for the analysis of HRV signal is linear analysis, and nonlinear analysis seldom used. Research progress of several methods of HRV analysis is presented. Principles and characteristics of using nonlinear analysis for the study of HRV are analyzed in detail. Fractal dimension, complexity and approximate entropy are discussed. PMID- 11892742 TI - The major subacrosomal occupant of bull spermatozoa is a novel histone H2B variant associated with the forming acrosome during spermiogenesis. AB - Recent studies on the structural composition of mammalian sperm heads have shown a congregate of unidentified proteins occupying the periphery of the mammalian sperm nucleus, forming a layer of condensed cytosol. These proteins are the perinuclear theca (PT) and can be categorized into SDS-soluble and SDS-insoluble components. The present study focused on identifying the major SDS-insoluble PT protein, which we localized to the subacrosomal layer of bovine spermatozoa and cloned by immunoscreening a bull testicular cDNA library. The isolated clones encode a protein of 122 amino acids that bears 67% similarity with histone H2B and contains a predicted histone fold motif. The novel amino terminus of the protein contains a potential bipartite nuclear targeting sequence. Hence, we identified this prominent subacrosomal component as a novel H2B variant, SubH2Bv. Northern blot analyses of SubH2Bv mRNA expression showed that it is testis specific and is also present in murid testes. Immunocytochemical analysis showed SubH2Bv intimately associates, temporally and spatially, with acrosome formation. While the molecular features of SubH2Bv are common to nuclear proteins, it is never seen developmentally within the nucleus of the spermatid. Considering its developmental and molecular characteristics, we have postulated roles of SubH2Bv in acrosome assembly and acrosome-nuclear docking. PMID- 11892743 TI - Effects of acute exposure to mild or moderate hypoxia on human psychomotor performance and visual-reaction time. AB - Objective. The aim of this study was to determine whether psychomotor performance and visual reaction time were affected by acute exposure to mild or moderate hypoxia. Method. Eighteen healthy male volunteers performed finger tapping, simple reaction time (SRT) and 4-choice reaction time (CRT) tests at simulated altitude of 300 m (control), 2800 m, 3600 m and 4400 m for 1 h in a hypobaric chamber. Result. SaO2 decreased from 98% (control) to 90%, 82% and 74% respectively at the various altitudes. All the performance parameters showed no significant change after exposure to 2800 m for 1 h relative to ground level (P>0.05). However the mean reaction time of 4-CRT under 3600 m prolonged and performance decreased as compared with baseline value (P<0.05), and the performance decreased further under 4400 m (P<0.01). No significant difference was found in finger tapping and SRT even under exposure to 4400 m for 1 h. Furthermore, no decrease in correct rate were observed at any altitude (P>0.05). Conclusion. The results from this study demonstrated that there were no measurable impairment of visual reaction time and psychomotor performance under exposure to an altitude of 2800 m for 1 h. However, adverse effects on psychomotor performance were observed under 3600 m and over. PMID- 11892744 TI - [Effects of sleep deprivation on human performance]. AB - Objective. To investigate the effects of sleep deprivation (SD) on human performance. Method. 8 healthy male college students participated the test. During 26 h of continuous awakeness (from 6:00 to 8:00 the next day), the volunteers were demanded to perform a battery of tests at 9 different time (7:00, 12:00, 16:00, 20:00, 0:00, 2:00, 4:00, 6:00, 8:00). The tests include: (1) single task: aural Oddball response, the response time (RT1) and correct rate (CR1) were recorded; (2) dual tasks: manual tracking and aural Oddball response, the response time (RT2), tracking error (ER) and correct rate (CR2) were recorded; (3) The Stanford sleepiness scale and subjective ratings of task difficulty access. Result. SD had significant effects on CT1, CT2 and ER (P=0.0001, P=0.00001, P=0.0004 respectively); SD increased RT1, RT2, ER at night time. SD had significant effects on SR, SSS score (P=0.0001, P=0.0000 respectively); SD increased SR, SSS score at night time. Since the subjects changed their response strategy, CR1 and CR2 were not influenced by SD at night time. Conclusion. SD has significant effects on response time, tracking error, subjective difficulty of cognitive tasks and subjective sleepiness. PMID- 11892745 TI - [Effects of tail suspension on learning and memory function of mice]. AB - Objective. To study the effects of simulated weightlessness on learning and memory capability of the brain. Method. Accuracy fraction, error frequency and pass rate were observed among control, restrained control, tail suspended (TS) control, restrained, and tail suspended mice in square water maze tests. And latent period and error time were observed in control and 30 degrees tail suspension mice in step down test. Result. The indices did not change significantly during learning period. Accuracy fraction of tail suspended group was reduced significantly in the tests as compared with pretest values. In step down test, latent period and error time showed no difference between TS 5 h and 2 d in untrained mice, but shortened evidently after TS 2 d and prolonged after TS 7 d in trained mice, which suggested the degradation of learning and memory ability after TS 5 h and 2 d respectively. Conclusion. Acute tail suspension depressed brain's learning ability and quality, while tail suspension of 7 d and 12 d impaired the spatial memory in mice. PMID- 11892746 TI - [Physiological evaluation of vestibular training load]. AB - Objective. To evaluate the effects of vestibular training by observing the variations of the physiological indices. It is helpful in grasping the training load, setting down and implementing the training plan. Method. 10 healthy subjects received linear acceleration, continuous Coriolis acceleration and discontinuous Coriolis acceleration stimuli on different dates. The stimulus was stopped when there were slight autonomic nervous symptoms, ECG, EGG and BP were recorded before during and after the stimuli. Computerized Dynamic Posturography (correction of Postrograph) (CDP) was tested before and after experiment. Result. One subject finished the training of the three stimuli with only slight autonomic nervous symptoms. The CDP tests pre- and post-experiment indicated that for most subjects the contribution of vestibular function in maintaining dynamic posture equilibrium increased after the linear acceleration stimuli, but decreased after continuous Coriolis acceleration stimuli, and there was great individual difference after discontinuous Coriolis acceleration stimuli because the stimulation was relatively heavy. The equilibrium score in SOT2 decreased significantly after linear acceleration stimuli, and increased significantly after discontinuous Coriolis acceleration stimuli. Conclusion. The training methods we designed and used in this experiment are feasible, and the required training load can be reached. EGG, BP and the percentage of LF (low frequency) in ECG R-R power spectrum can reflect the subject's condition when the period stimuli stopped. It provided important reference in the determination of training stimulation load. PMID- 11892747 TI - [A simulated study of effects of simulated hypovolemia on cardiovascular response to orthostatic stress]. AB - Objective. To study the effects of hypovolemia on cardiovascular response to orthostatic stress, and to investigate the role of hypovolemia in the mechanism of cardiovascular deconditioning and orthostatic intolerance induced by space weightlessness. Method. The effects of loss of blood volume had been incorporated in the sub-model of blood redistribution in the model developed for simulating the cardiovascular response to lower body negative pressure (LBNP). With the help of the model, we simulated the changes of heart rate (HR) and blood pressure (BP) during LBNP after 0-25% loss of blood volume. Result. When the amount of decrease of blood volume was less than 5% of the total blood volume, HR and BP could be maintained in normal range during LBNP through baroreflex regulation. When the amount of the decrease of blood volume was more than 15% of the total blood volume, HR and BP could be kept in normal range when the subject was supine and at rest. But BP fell sharply and the cardiovascular system almost collapsed during orthostatic exposure. Conclusion. Decrease of blood volume causes significant degradations of cardiovascular response to orthostatic stress. PMID- 11892748 TI - [A study of apoptosis and related gene bcl-2 and p53 expression in hippocampus of rats exposed to repeated +Gz]. AB - Objective. To investigate the role of apoptosis in mechanisms of brain damage induced by +Gz exposures. Method. Twenty conscious SD rats were randomly divided into 5 groups. Rats in the control group (n=4) were exposed to +1 Gz and rats in the 4 experimental groups (n=16) were exposed to +14 Gz for three times, each for 45 seconds with 30 min interval in between. All the +Gz exposured were on an animal centrifuge. The rat brains were taken 30 min, 6 h, 24 h and 48 h after the last centrifuge run and fixed and embedded. The apoptosis and expression changes of related gene bcl-2 and p53 were detected by terminal deoxynucleotide (correction of deoxynuleotide) transferase-mediated dUTP nick end labeling (TUNEL) technique and immunohistochemical method, respectively. Result. Apoptotic cells and expression changes of bcl-2 and p53 were observed in CA1 subregion of rat hippocampus taken 6 h after repeated +Gz exposures, but returned to normal after 24-48 h. Conclusion. It suggests that apoptosis and expression changes of bcl-2 and p53 in rats hippocampus can be induced by repeated +Gz exposures and the apoptosis is one of the molecular mechanisms of brain damage induced by repeated +Gz exposures. PMID- 11892749 TI - [Effects of CO2 concentration on growth and development of lettuce in controlled environment]. AB - Objective. To study the tolerance of lettuce to elevated CO2 concentration in Controlled Ecological Life Support System (CELSS). Method. Lettuce was cultivated in the Ground-based Experimental Facility for Higher Plant Cultivation in Space (GEFHPCS), in which many parameters were kept unchanged, while concentration of CO2 was controlled at 5 different levels (2000-10000 micromoles mol-1). During the growing periods, the morphologies of lettuce were observed every day, the replenished amounts of CO2 to GEFHPCS and water to the nutrient fluid box as well as the amounts of condensed water collected from GEFHPCS were all recorded every day. After harvest, the output and photosynthetic rate were calculated and lots of constituents of lettuce were analyzed. Result. The growth of lettuce were relatively ideal when CO2 concentration was at 6000 micromoles mol-1, but an obviously withering appearance was found when CO2 concentration increased 10000 micromoles mol-1, this time the output and quality of lettuce were unsatisfactory. Conclusion. It would be optimal when CO2 concentration is controlled at about 6000 micromoles mol-1 in a lettuce-cultivating chamber. PMID- 11892750 TI - [Design of reaction canister in a solid amine carbon dioxide removal system]. AB - Objective. To design a reaction canister using in solid amine carbon dioxide removal system for long-duration spaceflight. Method. On consideration of system demand and properties of solid amine, key problems must be solved were found out: 1) the rated resistance limit tends to shorten the length of the canister while absorption and concentration require to increase the length of the canister; 2) limited quantity of heat for keeping the temperature of the canister; 3) inflation or contraction of the solid amine under micro-gravity. Result. After appropriate measures were taken, effective adsorption and desorption, as well as concentration of CO2 were achieved, the concentration of CO2 in the space cabin could be controlled below 0.5%; and the concentration of the concentrated CO2 was as high as 98% so that it could be directly send to the CO2 reduction system; and that the resistance of the canister was below 160 mm H2O; moreover, the energy consumption was decreased to below 650 W. Conclusion. The designed reaction canister could meet the requirements of the solid amine carbon dioxide (correction of bioxide) removal system under microgravity. PMID- 11892751 TI - [A study of low temperature catalyst for Sabatier reaction]. AB - Objective. To develop a low temperature catalyst for the Sabatier CO2 reduction of the atmospheric regeneration system and lower the start-up temperature of the Sabatier reaction. Method. A low temperature catalyst was designed from the considerations of the active composition, the choice of the carrier, the production method and condition of the catalyst. Then the performance of the newly developed low temperature catalyst was tested. Result. A new low temperature catalyst for the Sabatier reaction using Ru as the active composition and using r-Al2O3 as the carrier was developed. The start-up temperature was lower than 110 degrees C and the start-up time was 8 min; The conversion efficiency of the lean component (H2 or CO2) was over 95 percent when the temperature of the reactor was from 200 degrees C to 300 degrees C; The reaction product water was nearly colorless, transparent and neutral. Conclusion. The test results showed that the goals of the design are achieved and it is worthwhile to make further studies on the low temperature catalyst. PMID- 11892752 TI - [Changes of femur minerals and serum BGP in hindlimb unloaded rats during convalescence]. AB - Objective. To observe bone mass changes during convalescence after simulated weightlessness. Method. 7-week-old rats were tail-suspended for 21 d then reloaded for 7 d and 21 d to recover, and measured serum BGP. Result. Tail suspension of rats for 21 d caused significant decrease of serum BGP and phosphorus as well as femur minerals. Serum BGP and femur minerals were still lower than control levels, but serum contents of calcium, phosphorus and magnesium increased significantly after reloading for 7 d. Femur minerals and serum BGP, calcium, phosphorus and magnesium returned to control levels after reloading for 21 d. Conclusion. The deficit in femur mineral induced by hindlimb unloading in rats can be restored by return to normal weight bearing, BGP can be used to monitor the case of its recovery. PMID- 11892753 TI - [Manual operation in manned spacecraft]. AB - The main character of manned spacecraft is that there are astronauts in it. In order to ensure security, reliability and high efficiency of the whole system, it is necessary to make full use of human operation in space environment at the stage of system overall design, especially for long manned flight. On the basis of related data, this paper summarized the principles of manual operation, its main objects and some requirement on man-machine interface. At last, some views have been put forward for discussion. PMID- 11892754 TI - [Change of pulmonary circulation in microgravity and simulated microgravity]. AB - Fluid is transferred cephalad in microgravity and simulated microgravity, and the pulmonary circulation is the first to be affected. Blood and fluid contents in the lungs increase, but unevenly distributed among various zones of the lungs. Blood vessels in the lungs are filled and distend. Capillary changes have been observed in the animal model of simulated microgravity. Studies about the changes of regulative function of pulmonary circulation are relatively rare. Observations of the reactivity of pulmonary vessels may help to understand the mechanisms of the changes in pulmonary circulation during microgravity. PMID- 11892755 TI - [Ultraviolet radiation and long term space flight]. AB - With the prolongation of space flight, influences of various aerospace environmental factors on the astronauts become more and more severe, while ultraviolet radiation is lacking. Some studies indicated that low doses of ultraviolet rays are useful and essential for human body. In space flight, ultraviolet rays can improve the hygienic condition in the space cabin, enhance astronaut's working ability and resistance to unfavorable factors, prevent mineral metabolic disorders, cure purulent skin diseases and deallergize the allergens. So in long-term space flight, moderate amount of ultraviolet rays in the space cabin would be beneficial. PMID- 11892756 TI - The association of hearing impairment and chronic diseases with psychosocial health status in older age. AB - OBJECTIVES: This study examines the association of hearing impairment and chronic diseases (diabetes mellitus, lung disease, cardiac disease, stroke, cancer, peripheral artery disease, osteoarthritis, rheumatoid arthritis) with psychosocial status (depression, self-efficacy, mastery, loneliness, social network size) in older persons. METHODS: The sample consists of 3,107 persons (55 to 85 years) participating in the Longitudinal Aging Study Amsterdam. MANOVA, adjusted for covariates, was used to test the effect of hearing impairment on the combined outcomes. The association of hearing impairment and chronic diseases with psychosocial status was studied using multivariate regression analyses. RESULTS: Hearing impaired elderly report significantly more depressive symptoms, lower self-efficacy and mastery, more feelings of loneliness, and a smaller social network than normally hearing peers. Whereas chronic diseases show significant associations with some outcomes, hearing impairment is significantly associated with all psychosocial variables. DISCUSSION: The findings emphasize the negative effect of hearing impairment on quality of life. PMID- 11892757 TI - Access to community-based long-term care: Medicaid's role. AB - OBJECTIVE: The authors explore state variation in expenditures for Medicaid community-based care services for the period 1990 to 1997. METHOD: A random effects panel model is used to explore the relationship between state demographic, supply, economic, programmatic, and political factors and states' Medicaid community-based care expenditures. RESULTS: Although states increased provision of services over the study period, significant state-level variation was evident. Expenditures were positively associated with state per capita income, regulation of nursing home bed supply, and the number of Medicare home health users but were negatively related to nursing home bed supply. CONCLUSIONS: Recent legal rulings, combined with the demonstrated preferences of most individuals to receive care in the community, require policies to foster the expansion of Medicaid community-based care. The most consistent relationships that are amenable to policy intervention relate to state fiscal resources and long-term care supply regulation. PMID- 11892758 TI - Development of a framework to encourage addressing advance directives when resources are limited. AB - OBJECTIVES: Advance directives are used to increase autonomy in decisions regarding care when a person lacks the ability to communicate such wishes. Based on studies showing internal consistency in individuals' preferences, this study demonstrates a new method for identifying a list of questions that may best predict patient preferences. METHOD: Participants were 71 residents of a large nursing home and 97 hospital patients. All were administered the Preferences for Life-Sustaining Treatment Questionnaire. Advance directives sequence models were developed using conditional probabilities concerning preferences for utilization of specific treatment options, given prior responses to different treatment options. RESULTS: Models resulted in more than 90% accuracy of treatment preferences for both samples and eliminated two thirds to three quarters of questions asked. CONCLUSIONS: Although the specific models need validation using larger samples, they demonstrate a method that facilitates transfer of information concerning treatment preferences if future incapacitation occurs. PMID- 11892759 TI - Service utilization among disabled Puerto Rican elders and their caregivers: does acculturation play a role? AB - OBJECTIVE: This study examined the relationship between language acculturation of disabled Puerto Rican elderly and their caregivers, their length of residence in mainland United States, and the utilization of formal services. METHODS: Language acculturation was measured by language use, understanding, and preferences. The sample of this study consisted of 194 dyads of disabled Puerto Rican elders 60 years and older, and their primary caregivers in an urban center in the northeast. RESULTS: Length of residence in the United States, but not language acculturation, of the disabled Puerto Rican elder and the caregiver was related to elder's use of formal services. Caregivers, whose own children were born in Puerto Rico as opposed to mainland United States, were more likely to use formal services. DISCUSSION: Language acculturation, although a commonly used measure of acculturation, may be of decreasing importance in explaining service utilization, as bilingual services become increasingly available. Practice implications are discussed. PMID- 11892760 TI - Death anxiety among Chinese elderly people in Hong Kong. AB - OBJECTIVES: This study aims to examine the cognitive and emotional reactions of Chinese elderly people toward death, to extend the use of a Western scale on death anxiety to a Chinese sample, and to explore the correlates of death anxiety. METHODS: A community sample of 237 Chinese elderly people (62 men and 175 women) in Hong Kong between the ages of 60 and 91 years old was individually interviewed. RESULTS: Among this elderly cohort, a high level of death anxiety was associated with younger age, a high level of psychological distress, and the presence of recent stressors but was unrelated to number of physical disorders, gender, personal income, marital and employment status, and religious affiliations. DISCUSSION: Reactions of Chinese elderly people toward specific death-related issues were discussed with regard to Chinese cultural beliefs. Limitations and implications of the present study were also discussed. PMID- 11892762 TI - Race, gender, and health care service utilization and costs among Medicare elderly with psychiatric diagnoses. AB - OBJECTIVE: To investigate race and gender differences in health care service utilization and costs among the Medicare elderly with psychiatric diagnoses. METHODS: The authors employ a 5% sample of Medicare beneficiaries from Tennessee (N = 33,680), and among those with a psychiatric diagnosis (n = 5,339), they examine health care service utilization and costs by race and gender. RESULTS: African Americans had significantly higher rates of diagnosis for dementia, organic psychosis, and schizophrenia, whereas Whites had significantly higher rates for mood and anxiety disorders. White and African American men have higher rates of utilization of emergency and inpatient services and lower rates of outpatient utilization compared to White women and African American women. African American men have significantly higher health care costs. DISCUSSION: The findings suggest that race and gender interact to influence service utilization and preventive care, thereby driving up costs of care, for elderly persons with psychiatric diagnoses. PMID- 11892761 TI - Disease prevention and health promotion in the aging with a traditional system of natural medicine: Maharishi Vedic Medicine. AB - OBJECTIVE: This review focuses on a comprehensive, sophisticated system of natural medicine that appears to hold promise for prevention of chronic diseases and disabilities, loss of independence, suffering, and health care costs often associated with "usual" aging. METHODS: The authors discuss the negative impact of usual aging on our society, with its rapidly growing percentage of elderly, and the challenge of promoting "successful aging." Emphasis is given to research literature suggesting that Maharishi Vedic Medicine (MVM) is particularly effective in retarding usual aging. RESULTS: Proposed mechanisms for the antiaging effects of MVM include reductions in physiological and psychological stress and enhancement of homeostatic and self-repair processes. CONCLUSIONS: The authors conclude that this set of innovative strategies may help society achieve recommended health objectives for disease prevention and health promotion in older adults and that widespread implementation of this self-empowering, prevention-oriented approach in the elderly is feasible, cost effective, and timely. PMID- 11892763 TI - Health of grandmothers: a comparison by caregiver status. AB - OBJECTIVES: In this cross-sectional study, stress, coping, social support, and health were compared in 86 primary caregiver grandmothers, 85 partial/supplemental caregiver grandmothers in multigenerational homes, and 112 noncaregiver grandmothers. Whether support and coping reduced effects of stress on the physical and mental health of grandmother caregivers, including mediating and moderating effects, was examined. METHODS: A convenience sample completed a mailed questionnaire that included measures of stress, health, support, and coping. RESULTS: Primary caregivers reported worse self-assessed health, but partial/supplemental caregivers reported a tendency toward more depression and more instrumental support. Noncaregivers reported the least stress and less active and avoidant coping. Coping and subjective support added to the variance of depression and self-rated health. Subjective support and avoidant coping mediated between stress and health, and active coping moderated the effects of stress on health. DISCUSSION: Implications of these findings relative to the health of grandmothers by caregiver status are discussed. PMID- 11892764 TI - Inequitable access: Medicare+Choice program fails to serve rural America. PMID- 11892765 TI - Comments on regulatory and contractor reform legislation. PMID- 11892767 TI - Board to review modifications to sentinel event procedures. PMID- 11892769 TI - Board votes to increase time frame for submitting root cause analysis. PMID- 11892768 TI - Board of Commissioners affirms support for Sentinel Event Policy. PMID- 11892770 TI - Lessons learned: wrong site surgery. PMID- 11892771 TI - Inpatient suicides: recommendations for prevention. PMID- 11892772 TI - Enjoying goal-directed action: the role of regulatory fit. AB - We propose that the fit between an action's strategic orientation and the actor's regulatory state can influence the amount of enjoyment the action provides. In two studies using different methods of manipulating regulatory states and of gauging action evaluations, high regulatory fit increased participants' anticipations of action enjoyability. In a third study, high regulatory fit increased participants' enjoyment of perceived success at, and willingness to repeat a novel laboratory task, and these effects were independent of participants' actual success on the task. Across the three studies, participants in a regulatory state oriented toward accomplishment experienced eagerness related actions more favorably than vigilance-related actions, whereas participants in a regulatory state oriented toward responsibility experienced vigilance-related actions more favorably than eagerness-related actions. These findings' implications for understanding task interest and motivation are discussed. PMID- 11892773 TI - Object name learning provides on-the-job training for attention. AB - By the age of 3, children easily learn to name new objects, extending new names for unfamiliar objects by similarity in shape. Two experiments tested the proposal that experience in learning object names tunes children's attention to the properties relevant for naming--in the present case, to the property of shape -and thus facilitates the learning of more object names. In Experiment 1, a 9 week longitudinal study, 17-month-old children who repeatedly played with and heard names for members of unfamiliar object categories well organized by shapeformed the generalization that only objects with ith similar shapes have the same name. Trained children also showed a dramatic increase in acquisition of new object names outside of the laboratory during the course of the study. Experiment 2 replicated these findings and showed that they depended on children's learning both a coherent category structure and object names. Thus, children who learn specific names for specific things in categories with a common organizing property--in this case, shape--also learn to attend to just the right property- in this case, shape--for learning more object names. PMID- 11892774 TI - Where did 1850 happen first--in America or in Europe? A cognitive account for a historical bias. AB - A professor of history at The Hebrew University noted that his students were often surprised to learn that some event in America happened at about the same time as another in Europe, because the American event seemed to them to have happened more recently. We confirmed the validity, of this anecdotal observation experimentally, and offer an explanation. We discuss how this bias may be an effect of judgment, rather than memory. We then show experimentally that students like those who demonstrated the bias regarded America as the New World, as opposed to Europe's Old World. Our theoretical account, based on judgment by representativeness, posits that if one category is deemed more X than another (e.g., American history is deemed more "recent" than European history), then its members will be judged more X than members of the other, ceteris paribus. Hence, an American historical event will appear more recent than a contemporaneous European event. PMID- 11892775 TI - Forgetting to remember: the functional relationship of decay and interference. AB - Functional decay theory proposes that decay and interference, historically viewed as competing accounts of forgetting, are instead functionally related. The theory posits that (a) when an attribute must be updated frequently in memory, its current value decays to prevent interference with later values, and (b) the decay rate adapts to the rate of memory updates. Behavioral predictions of the theory were tested in a task-switching paradigm in which memory for the current task had to be updated every few seconds, hundreds of times. Reaction times and error rates both increased gradually between updates, reflecting decay of memory for the current task. This performance decline was slower when updates were less frequent, reflecting a decrease in the decay rate following a decrease in the update rate. A candidate mechanism for controlled decay is proposed, the data are reconciled with practice effects, and implications for models of executive control are discussed. PMID- 11892776 TI - Attending to the big picture: mood and global versus local processing of visual information. AB - Two experiments employed image-based tasks to test the hypothesis that happier moods promote a greater focus on the forest and sadder moods a greater focus on the trees. The hypothesis was based on the idea that in task situations, affective cues may be experienced as task-relevant information, which then influences global versus local attention. Using a serial-reproduction paradigm, Experiment 1 showed that individuals in sad moods were less likely than those in happier moods to use an accessible global concept to guide attempts to reproduce a drawing from memory. Experiment 2 investigated the same hypothesis by assessing the use of global and local attributes to classify geometricfigures. As predicted, individuals in sad moods were less likely than those in happier moods to classify figures on the basis of globalfeatures. PMID- 11892777 TI - Influence of past experience on perceptual grouping. AB - We used primed matching to examine the microgenesis of perceptual organization for familiar (upright letters) and unfamiliar (inverted letters) visual configurations that varied in the connectedness between their line components. The configurations of upright letters were available for priming as early as 40 ms, irrespective of connectedness between their line components. The configurations of connected inverted-letter primes were also available this early, but the configurations of disconnected inverted letters were not available until later These results show that past experience contributes to the early grouping of disconnected line segments into configurations. These findings suggest an interactive model of perceptual organization in which both image-based properties (e.g., connectedness) and input from object memories contribute to perceptual organization. PMID- 11892778 TI - As seen by the other...: Perspectives on the self in the memories and emotional perceptions of Easterners and Westerners. AB - The experiment reported investigated the phenomenological consequences of Easterners' and Westerners'perspectives on the self Two findings are consistent with the notion that Asians are more likely than Westerners to experience the self from the perspective of the generalized other First, Eastern participants were more likely than Western participants to have third-person (as opposed to first-person) memories when they thought about situations in which they would be at the center of a scene. Second, Easterners and Westerners engaged in different sorts of projections when they read the emotional expressions of other people. Westerners were more biased than Easterners toward egocentric projection of their own emotions onto others, whereas Easterners were more biased than Westerners toward relational projection, in which they projected onto others the emotions that the generalized other would feel in relation to the participant. Implications for how phenomenological experiences could reinforce different Eastern and Western ideologies about the self and the group are discussed. PMID- 11892780 TI - Second thoughts versus second looks: an age-related deficit in reflectively refreshing just-activated information. AB - Age-related deficits in memory are greater as encoding and retrieval tasks require more reflective (self-generated or executive) processing. One problem in developing more specific models of age-related changes in cognition is that the tasks studied tend to be complex and vary in the combinations of component cognitive processes they recruit. Here we report an age-related deficit in one of the most elementary, but critical, components of reflection: refreshing a just activated representation. Impairment in such a process potentially has a wide ranging impact on all higher-order cognition. PMID- 11892779 TI - Chimpanzees (Pan troglodytes) recognize spatial and object correspondences between a scale model and its referent. AB - In the present study, the contributions of spatial and object features to chimpanzees' comprehension of scale models were examined. Seven chimpanzees that previously demonstrated the ability to use a scale model as an information source for the location of a hidden item were tested under conditions manipulating the feature correspondence and spatial-relational correspondence between objects in the model and an outdoor enclosure. In Experiment 1, subjects solved the task under two conditions in which one object cue (color or shape) was unavailable, but positional cues remained. Additionally, performance was above chance under a third condition in which both types of object cues, but not position cues, were available. In Experiment 2, 2 subjects solved the task under a condition in which shape and color object cues were simultaneously unavailable. The results suggest that, much like young children, chimpanzees are sensitive to both object and spatial-relational correspondences between a model and its referent. PMID- 11892781 TI - Caffeine reduces time-of-day effects on memory performance in older adults. AB - For most older adults, memory performance depends on the time of day, with performance being optimal early in the morning and declining during the late afternoon hours. In the present study, we asked whether this decline could be ameliorated by a simple stimulant, caffeine. Adults over the age of 65 who considered themselves "morning types" were tested twice over an interval of 5 to 11 days, once in the morning and once in the late afternoon. Participants ingested either coffee with caffeine or decaffeinated coffee at both sessions. Participants who ingested decaffeinated coffee showed a significant decline in memory performance from morning to afternoon. In contrast, those who ingested caffeine showed no decline in performance from morning to afternoon. The results suggest that time-of-day effects may be mediated by nonspecific changes in level of arousal. PMID- 11892782 TI - Sexual and romantic jealousy in heterosexual and homosexual adults. AB - Several theorists have claimed that men are innately more upset by a mate's sexual infidelity and women are more upset by a mate's emotional infidelity because the sexes faced different adaptive problems (for men, cuckoldry; for women, losing a mate's resources). The present work examined this theory of jealousy as a specific innate module in 196 adult men and women of homosexual and heterosexual orientations. As in previous work, heterosexuals' responses to a forced-choice question about hypothetical infidelity yielded a gender difference. However no gender differences were found when participants recalled personal experiences with a mate's actual infidelity. Men and women, regardless of sexual orientation, on average focused more on a mate's emotional infidelity than on a mate's sexual infidelity. Responses to hypothetical infidelity were uncorrelated with reactions to actual infidelity. This finding casts doubt on the validity of the hypothetical measures used in previous research. PMID- 11892783 TI - Absolute pitch and tempo in mothers' songs to infants. AB - We examined the relative stability of pitch, tempo, and rhythm in maternal speech and singing to prelinguistic infants. Mothers were recorded speaking and singing to their infants on two occasions separated by 1 week or more. The pitch level and tempo of identical utterances were highly variable across the 1-week period, but these features were virtually unchanged in song repetitions. Rhythmic patterning was largely maintained in speech, as in song. Mothers' accurate reproduction of their sung performances can be considered a form of absolute pitch and absolute tempo. PMID- 11892784 TI - Color-based motion processing is stronger in infants than in adults. AB - One hallmark of vision in adults is the dichotomy between color and motion processing. Specifically, areas of the brain that encode an object's direction of motion are thought to receive little information about object color We investigated the development of this dichotomy by conducting psychophysical experiments with human subjects (2-, 3-, and 4-month-olds and adults), using a novel red-green stimulus that isolates color-based input to motion processing. When performance on this red-green motion stimulus was quantified with respect to performance on a luminance (yellow-black) standard, we found stronger color-based motion processing in infants than in adults. These results suggest that color input to motion areas is greater early in life, and that motion areas then specialize to the adultlike state by reweighting or selectively pruning their inputs over the course of development. PMID- 11892785 TI - The affective consequences of expected and unexpected outcomes. AB - How do people feel about unexpected positive and negative outcomes? Decision affect theory (DAT) proposes that people feel displeasure when their outcomes fall short of the counterfactual alternative and elated when their outcomes exceed the counterfactual alternative. Because disconfirmed expectations provide a counterfactual alternative, DAT predicts that bad outcomes feel worse when unexpected than when expected, yet good outcomes feel better when unexpected than when expected. Consistency theories propose that people experience displeasure when their expectations are disconfirmed because the disconfirmation suggests an inability to predict. According to consistency theories, both good and bad outcomes feel worse when unexpected than when expected. These two theoretical approaches were tested in three studies. The results consistently support DAT PMID- 11892786 TI - Perceptual organization overcomes the effects of local surround in determining simultaneous lightness contrast. AB - Lightness induction can occur on the basis of the immediate surround of a region (local interactions) and also on the basis of global factors of perceptual organization. The experiments reported in this article used novel displays that made it possible to differentiate the contributions of these two kinds offactors. The experiments demonstrated, for the first time, that when higher-level factors act contemporaneously with lower-level factors, the contrast effect induced by the global-organization principle of perceptual belongingness overcomes the effect due to retinal lateral inhibition. PMID- 11892787 TI - Intramolecular binding of a proximal PPII helix to an SH3 domain in the fusion protein SH3Hck : PPIIhGAP. AB - SH3 domains are a conserved feature of many nonreceptor protein tyrosine kinases, such as Hck, and often function in substrate recruitment and regulation of kinase activity. SH3 domains modulate kinase activity by binding to polyproline helices (PPII helix) either intramolecularly or in target proteins. The preponderance of bimolecular and distal interactions between SH3 domains and PPII helices led us to investigate whether proximal placement of a PPII helix relative to an SH3 domain would result in tight, intramolecular binding. We have fused the PPII helix region of human GAP to the C-terminus of Hck SH3 and expressed the recombinant fusion protein in Escherichia coli. The fusion protein, SH3Hck : PPIIhGAP, folded spontaneously into a structure in which the PPII helix was bound intramolecularly to the hydrophobic crevice of the SH3 domain. The SH3Hck : PPIIhGAP fusion protein is useful for investigating SH3: PPII helix interactions, for studying concepts in protein folding and design, and may represent a protein structural motif that is widely distributed in nature. PMID- 11892788 TI - Implications of SH3 domain structure and dynamics for protein regulation and drug design. AB - SH3 Domains provide interesting targets for investigations of protein structure and dynamics because of their compact size and importance for signal transduction. The present review summarizes recent research investigating SH3 domain structure and dynamics, the discovery of novel SH3 domains, the role of SH3 domains in disease, and progress in targeting SH3 domains for the development of novel therapeutics. Particular emphasis is placed on the unfolding/refolding characteristics of SH3 domains and the potential importance of these processes for regulation of signal transduction. PMID- 11892789 TI - Repair of oxidative DNA damage: mechanisms and functions. AB - Cellular genomes suffer extensive damage from exogenous agents and reactive oxygen species formed during normal metabolism. The MutT homologs (MutT/MTH) remove oxidized nucleotide precursors so that they cannot be incorporated into DNA during replication. Among many repair pathways, the base excision repair (BER) pathway is the most important cellular protection mechanism responding to oxidative DNA damage. The 8-oxoG glycosylases (Fpg or MutM/OGG) and the MutY homologs (MutY/MYH) glycosylases along with MutT/MTH protect cells from the mutagenic effects of 8-oxoG, the most stable and deleterious product known caused by oxidative damage to DNA. The key enzymes in the BER process are DNA glycosylases, which remove different damaged bases by cleavage of the N glycosylic bonds between the bases and the deoxyribose moieties of the nucleotide residues. Biochemical and structural studies have demonstrated the substrate recognition and reaction mechanism of BER enzymes. Cocrystal structures of several glycosylases show that the substrate base flips out of the sharply bent DNA helix and the minor groove is widened to be accessed by the glycosylases. To complete the repair after glycosylase action, the apurinic/apyrimidinic (AP) site is further processed by an incision step, DNA synthesis, an excision step, and DNA ligation through two alternative pathways. The short-patch BER (1-nucleotide patch size) and long-patch BER (2-6-nucleotide patch size) pathways need AP endonuclease to generate a 3' hydroxyl group but require different sets of enzymes for DNA synthesis and ligation. Protein-protein interactions have been reported among the enzymes involved in BER. It is possible that the successive players in the repair pathway are assembled in a complex to perform concerted actions. The BER pathways are proposed to protect cells and organisms from mutagenesis and carcinogenesis. PMID- 11892790 TI - Slow inactivation in voltage-gated sodium channels: molecular substrates and contributions to channelopathies. AB - Slow inactivation in voltage-gated sodium channels is a biophysical process that governs the availability of sodium channels over extended periods of time. Slow inactivation, therefore, plays an important role in controlling membrane excitability, firing properties, and spike frequency adaptation. Defective slow inactivation is associated with several diseases of cell excitability, such as hyperkalemic periodic paralysis, myotonia, idiopathic ventricular fibrillation and long-QT syndrome. These associations underscore the physiological importance of this phenomenon. Nevertheless, our understanding of the molecular substrates for slow inactivation is still fragmentary. This review covers the current state of knowledge concerning the molecular underpinnings of slow inactivation, and its relationship with other biophysical processes of voltage-gated sodium channels. PMID- 11892791 TI - Signal integration and the specificity of insulin action. AB - Insulin is a potent metabolic hormone essential for the maintenance of normal circulating blood glucose level in mammals. The physiologic control of glucose homeostasis results from a balance between hepatic glucose release (glycogenolysis and gluconeogenesis) and dietary glucose absorption versus skeletal muscle and adipose tissue glucose uptake and disposal. Disruption of this delicate balance either through defects in insulin secretion, liver glucose output, or peripheral tissue glucose uptake results in pathophysiological states of insulin resistance and diabetes. In particular, glucose transport into skeletal muscle and adipose tissue is the rate-limiting step in glucose metabolism and reduction in the efficiency of this process (insulin resistance) is one of the earliest predictors for the development of Type II diabetes. Importantly, recent studies have directly implicated an impairment in insulin receptor signal transduction as the prime mechanism for peripheral tissue insulin resistance. In this review, we have focused on recent developments in our understanding of the molecular mechanisms and signal transduction pathways that insulin utilizes to specifically regulate glucose uptake. The detailed understanding of these events will provide a conceptual framework for the development of new therapeutic targets to treat this chronic and debilitating disease process. PMID- 11892792 TI - Coordinately expressed chorion genes of Bombyx mori: is developmental specificity determined by secondary structure recognition? AB - Short inverted repeat sequences have been observed in the DNA flanking the 5' and 3' termini of a pair of coordinately expressed chorion structural genes of the silkmoth Bombyx mori. When superhelical cloned DNA containing the two chorion genes is digested with S1 nuclease, several sites are specifically cleaved including those around the centers of the putative cruciform structures resulting from the short inverted repeat sequences. The possible implication of conformational changes in the DNA surrounding the chorion structural genes in the process of determination of the developmental specificity and the coordinate transcriptional regulation of these genes is discussed. PMID- 11892793 TI - X-ray absorption near edge structure (XANES) for CO, CN and deoxyhaemoglobin: geometrical information. AB - We use the recently developed multiple scattering theory to give a quantitative analysis of the X-ray absorption near edge structure (XANES) of haemoglobin and some of its substituents. We demonstrate that the XANES may contain information not provided by the extended X-ray absorption fine structure (EXAFS) part of the spectrum about the coordination geometry around the Fe atom, and in particular discuss the sensitivity of the XANES to the orientation of the CN group in HbCN. The anisotropy of the system leads to a strong dependence of the calculated spectrum on the polarisation of the X-rays. We show how this effect can be exploited in further XANES structural studies. PMID- 11892794 TI - Developmental changes in translatable mRNAs for the cerebral enolase isozymes alphaalpha and gammagamma. AB - Using the rabbit reticulocyte cell-free translation system we have estimated during ontogenesis the proportions of in vitro translatable alpha and gamma brain enolase mRNAs, which are two minor mRNA species. No polypeptide precursor to these enzyme subunits appears to be synthesized during translation in vitro. During brain development, the changes in translatable alpha and gamma mRNA content seem to parallel those of the corresponding antigens. The proportion of each of the enolase mRNAs is highest in adult mouse brain. Mechanisms controlling alpha and gamma antigen expression are discussed. In order to prepare the specific cDNA probes, purification of alpha and gamma mRNAs was undertaken. PMID- 11892795 TI - Biosynthesis of the sperm receptor during oogenesis in the mouse. AB - During their growth phase, mouse oocytes synthesize and secrete three different glycoproteins, called ZP1, 2 and 3, that constitute the extracellular coat, or zona pellucida, of the oocyte. One of these glycoproteins, ZP3, exhibits properties expected for a sperm receptor. We have now used rabbit antisera that recognize ZP3 to immunoprecipitate [35S]methionine-labeled, intracellular precursors of this glycoprotein from growing oocytes cultured in vitro in the presence or absence of tunicamycin, a drug that prevents addition of N-linked oligosaccharides to nascent polypeptide chains. Electrophoretic analyses of these immunoprecipitates, as well as of immunoprecipitates digested with endo-beta-N acetylglucosaminidase H (Endo H), indicate that ZP3 is synthesized as a 44,000 mol. wt. polypeptide chain to which either three or four high-mannose-type oligosaccharides are added, resulting in 53,000 and 56,000 mol. wt. ZP3 precursors, respectively. The latter species are converted to mature ZP3 (mol. wt. approximately 80,000) by processing of the high-mannose-type oligosaccharides (Endo H-sensitive) to complex-type oligosaccharides (Endo H-insensitive) prior to ZP3 secretion. The evidence presented reveals that the extreme heterogeneity of mature ZP3, with respect to both mol. wt. and isoelectric point, is partly a consequence of the N-linked oligosaccharides and not the polypeptide chain itself. PMID- 11892796 TI - Monoclonal antibodies displaying a novel species specificity for the primate transformation-related protein, p53. AB - SV40 large T antigen associates with a cellular phosphoprotein, p53, in virus transformed cells. We have raised three new monoclonal antibodies, PAb1101, PAb1102 and PAb1103, to this cellular protein, derived from SV40-transformed human fibroblasts. These define at least two non-overlapping determinants on human p53 that are in different areas of the molecule from those recognised by previously available antibodies. Unlike those antibodies, PAb1102 and PAb1103 do not react with rodent p53. PAb1101 reacts far more weakly with rodent p53 than with primate p53. All three antibodies show a preference for binding to the large T-associated form of p53, an effect that is particularly marked with PAb1102. The novel specificity of these antibodies allows further probing of the nature and function of the large T/p53 complex in human cells. PMID- 11892797 TI - Map location of transcripts from Torulopsis glabrata mitochondrial DNA. AB - Unique transcripts for cytochrome b, ATPase subunits 6 and 9, cytochrome oxidase subunits 2 and 3 and S and L rRNA have been mapped by the S1 protection technique to the circular 19-kbp mitochondrial DNA (mtDNA) of the yeast Torulopsis glabrata. In contrast, a number of transcripts have been detected for the mosaic cytochrome oxidase subunit 1 gene with the largest being approximately 5000 nucleotides and the mature message having a length of 1760 nucleotides. Despite the presence in T. glabrata mtDNA of a sequence that hybridizes to the variant 1 gene of Saccharomyces cerevisiae mtDNA we have not detected a transcript of this region. Neither have we detected co-transcripts of adjacent genes in RNA from either glucose-repressed or derepressed cells. However, by comparison of RNA species from the two growth conditions, we have found that the ATPase subunit 6 transcript is lower in amount relative to other species in preparations from glucose-repressed cells. This information, together with the observation of separate transcripts and the knowledge that there are several species of mitochondrial RNA which can be capped by the guanylyl transferase catalysed addition of GMP, suggests that each of the genes investigated in the present study is separately transcribed. PMID- 11892798 TI - Two specific markers for neural differentiation of embryonal carcinoma cells. AB - Two multipotential embryonal carcinoma (EC) cell lines, 1003 and 1009, can be induced to form preferentially neural derivatives in vitro. Synthesis of specific proteins during neural differentiation was followed by two-dimensional gel electrophoresis. The comparison of protein patterns obtained with neural and non neural derivatives of these EC cell lines indicates that two changes are specific for the neural pathway: (i) the appearance of a new beta-tubulin isoform and (ii) the accumulation of the brain isozyme of creatine phosphokinase already present in small amounts in EC stem cells. These changes were found to take place early in the course of differentiation and to occur even when neurite outgrowth was prevented. PMID- 11892800 TI - A bovine papilloma virus vector with a dominant resistance marker replicates extrachromosomally in mouse and E. coli cells. AB - We describe the construction of a bovine papilloma virus-based vector (pCGBPV9) which contains a dominant selectable marker and replicates autonomously in both mouse and Escherichia coli cells. This vector contains the complete bovine papilloma virus genome, a ColE1 replication origin and a dominant selectable marker conferring resistance to kanamycin in bacteria and G418 in eukaryotic cells. A high number of G418R colonies are obtained after transfer of pCGBPV9 into mouse C127 cells. These G418R colonies contain vector DNA which replicates autonomously at approximately 10-30 copies per cell. The molecules are in most cases unrearranged and can be rescued into E. coli cells by bacterial transformation. PMID- 11892799 TI - Analysis of vertebrate gap junction protein. AB - A new method for the purification of gap junctions is described which depends on the extraction of cell monolayers or tissue homogenates with Triton X-100. The major band on SDS-polyacrylamide gel electrophoresis (PAGE) of junctional preparations from a variety of vertebrate sources has an apparent mol. wt. of 16,000 (16 K). Further evidence for the junctional origin of the 16 K protein is provided by the results of four different experimental approaches. (i) The junctions form a sharp band in potassium iodide density gradients at 1.195 g/cm3 and the 16 K protein is the only detectable band in fractions of this bouyant density. (ii) The junctions are progressively solubilised by increasing concentrations of SDS (in the range 0.1-0.5%) and the dissolution of the junctional structure, observed by electron microscopy, parallels the release of the 16 K protein. (iii) Glutaraldehyde fixation of intact junctions cross-links the 16 K protein. (iv) The recoverable amount of the 16 K protein correlates with known changes in gap junctional area in the regenerating weanling rat liver after partial hepatectomy and in V79 cell cultures exposed to 4beta-phorbol 12 myristate 13-acetate. PMID- 11892802 TI - Isolation, molecular and functional properties of the C-terminal domain of colicin A. AB - Partial proteolytic digestion of colicin A with bromelain allowed the isolation of a 20-kd fragment. This fragment has been purified to homogeneity and its molecular properties have been studied. The sequence of the 54 N-terminal amino acid residues has been determined by automated Edman degradation. This sequence is identical to that of the predicted amino acid sequence of the 20-kd C-terminal part of the colicin A polypeptide deduced from the nucleotide sequence of the caa gene. This polypeptide can produce channels in phospholipid planar bilayers of the same size as those formed by colicin A. However, the voltage-dependence for opening and closing was drastically altered in the peptide fragment channels. The latter, in contrast to colicin A channels, remained open over a wide range of voltage. Large negative potentials were required to close the peptide fragment channels although opening took place in the same voltage range as for colicin A ionic pores. PMID- 11892801 TI - Inverse control of prolactin and growth hormone gene expression: effect of thyroliberin on transcription and RNA stabilization. AB - The hypothalamic tripeptide thyroliberin (TRH) regulates prolactin (PRL) and growth hormone (GH) synthesis inversely by modulating the levels of their specific mRNA. Changes in mRNA levels could involve both transcriptional and posttranscriptional events. To examine further these possibilities, we have investigated the effect of TRH on the biosynthesis and degradation of PRL and GH RNA in a rat pituitary tumor cell line. Newly synthesized PRL and GH RNA sequences were quantified in nuclear and cytoplasmic fractions by hybridization of 3H-labelled RNA to immobilized plasmid DNA containing either PRL or GH cDNA sequences. Steady-state levels of specific RNA were estimated by RNA blot hybridization. The results indicate that TRH increases in a rapid but transient manner the transcription of the PRL gene, and suggest that it does not alter the processing and the transport to the cytoplasm. In contrast, after a lag-time, TRH seems to induce a long-lasting inhibition on GH, as well as on overall gene transcription. Furthermore, we observed an effect of TRH on mRNA stability. TRH significantly increases the half-life of PRL mRNA. Our results also support the hypothesis that TRH decreases the half-life of GH mRNA. Such post-transcriptional action of TRH amplifies and prolongs the regulations exerted at the transcriptional level. PMID- 11892803 TI - Alteration of vimentin intermediate filament expression during differentiation of human hemopoietic cells. AB - We describe the alterations of vimentin intermediate filament (IF) expression in human hemopoietic committed precursors as they differentiate into mature cells of the erythroid, granulomonocytic, megacaryocytic and lymphoid lineages. A double labelling fluorescence procedure was used to identify hemopoietic cells expressing lineage-specific antigens and to decorate the vimentin IF network. Whereas very early progenitors from each lineage expressed vimentin, the density and organization of the network differed strikingly as the cells matured on a given pathway. T lymphocytes, monocytes and granulocytes retained vimentin expression at all stages of maturation. In contrast, megakaryoblasts lose vimentin expression at a very early stage of differentiation, erythroblasts at variable steps between the committed erythroid cell and the red cell. Finally, B lymphocytes tend to lose vimentin expression later when they mature into plasma cells. PMID- 11892804 TI - Accuracy of DNA polymerase-alpha in copying natural DNA. AB - The fidelity of DNA polymerase-alpha from calf thymus (9S enzyme) in copying bacteriophage phi174am16 DNA in vitro has been determined from the frequency of production of different revertants. In the self-priming reaction we were able to measure the frequencies of base pairing mismatches during the course of replication on biasing the ratios of deoxynucleoside triphosphates. The frequency of dGTP:T, dGTP:G and dATP:G mismatches were 7.6 x 10(-5), 4.4 x 10(-5) and 2.8 x 10(-5), respectively, at equal concentrations of the deoxynucleoside triphosphates. dCTP:A, dGTP:A, dCTP:T and dTTP:T mismatches were below the limit of detection (<5 x 10(-6)). A synthetic dodecamer primer with a 3' end covering the first two bases of the amber codon was used to determine the misinsertion frequency of the first nucleotide incorporated. This gave a misinsertion frequency of 1.5 x 10(-4) for the dGTP:T mismatch, which is slightly higher than that observed from the pool bias studies. Further, it showed no sensitivity to biasing the nucleotide pool, suggesting a different mechanism for the incorporation of the first nucleotide. These data do not support 'energy-relay' like models for achieving high accuracy in eukaryotes. The observed misinsertion frequencies were corrected for mismatch repair of the heteroduplexes during the transfection experiments by parallel experiments using a mismatched primer. This was synthesized to have the same G:T mismatch as produced in the preceding experiment. PMID- 11892805 TI - The lysis function of RNA bacteriophage Qbeta is mediated by the maturation (A2) protein. AB - Complete or partial cDNA sequences of the RNA bacteriophage Qbeta were cloned in plasmids under the control of the lambdaP(L) promoter to allow regulated expression in Escherichia coli harbouring the gene for the temperature-sensitive lambdaCI857 repressor. Induction of the complete Qbeta sequence leads to a 100 fold increase in phage production, accompanied by cell lysis. Induction of the 5' terminal sequence containing the intact maturation protein (A2) cistron also causes cell lysis. Alterations of the A2 cistron, leading to proteins either devoid of approximately 20% of the C-terminal region or of six internal amino acids, abolish the lysis function. Expression of other cistrons in addition to the A2 cistron does not enhance host lysis. Thus, in Qbeta, the A2 protein, in addition to its functions as maturation protein, appears to trigger cell lysis. This contrasts with the situation in the distantly related group I RNA phages such as f2 and MS2 where a small lysis polypeptide is coded for by a region overlapping the end of the coat gene and the beginning of the replicase gene. PMID- 11892806 TI - Monoclonal antibodies to human interferon-gamma: production, affinity purification and radioimmunoassay. AB - Human interferon-gamma (IFN-gamma) purified to electrophoretic homogeneity by a cation exchange h.p.l.c., was used for the development of monoclonal antibodies. Following immunization, spleen lymphocytes of two mice showing the highest binding and neutralizing titers were isolated, fused with NSO mouse myeloma cells and cloned. The screening of hybridomas was based on precipitation of the immune complexes with a second antibody and recovery of the biological activity of IFN gamma from the precipitate. Twenty nine independent hybridomas secreting antibodies specific to IFN-gamma were obtained. Twelve out of these 29 hybridomas produced antibodies that neutralized the antiviral activity of pure as well as crude IFN-gamma. Moreover, IFN-gamma obtained by various induction procedures was neutralized as well, indicating that these various IFN-gamma subtypes are immunologically cross-reactive. Immune precipitation of partially purified 125I labelled IFN-gamma by several monoclonal antibodies revealed two protein bands of 26,000 and 21,000 daltons. Immunoaffinity chromatography of IFN-gamma gave a 50 fold purification to a specific activity > or = 4 x 10(7) units/mg. Two of the monoclonal antibodies were found suitable for a sensitive and rapid double antibody solid-phase radioimmunoassay, allowing the detection of IFN-gamma at concentrations of at least 4 ng/ml (150 units/ml) within 8 h. PMID- 11892807 TI - Torsional stress induces left-handed helical stretches in DNA of natural base sequence: circular dichroism and antibody binding. AB - Above a threshold of torsional stress, the c.d. spectrum of covalently closed circular DNA of natural base sequence acquires a Z-like contribution and antibodies raised against Z-DNA are bound. Mapping of the antibody binding sites by electron microscopy reveals sites which correlate with stretches enriched in alternating purine-pyrimidine sequences and GC base pairs. PMID- 11892808 TI - H-2 hemizygous mutants from a heterozygous cell line: role of mitotic recombination. AB - Variants that no longer express an entire H-2 haplotype were readily isolated, by immunoselection with antisera directed against the haplotype, from an H-2b/H-2d heterozygous Friend leukemia cell line carrying a Robertsonian translocation of the chromosomes bearing the H-2 genetic region. These variants can be denoted as being of the phenotype H-2b- H-2d+ or H-2b+ H-2d-. Some of the H-2b- H-2d+ variants: (1) lack the restriction enzyme fragments characteristic of the missing H-2b haplotype, as assessed by Southern blot analysis; (2) express more cell surface H-2d antigens than wild-type cells, as assessed by flow microfluorimetry; and (3) appear to have become homozygous for the more active H-2d-linked allele at the Glyoxalase I locus. These variants thus seem to have lost genetic material corresponding to the H-2b haplotype and may have gained genetic material corresponding to the H-2d haplotype. These results are consistent with the possibility that these variants were generated by mitotic recombination. PMID- 11892810 TI - Subcellular localization of viroids in highly purified nuclei from tomato leaf tissue. AB - Approximately 95% of the viroid RNA which is present in potato spindle tuber viroid (PSTV)-infected tomato plant leaf issue, is associated with the nucleolar fraction obtained from purified nuclei. Viroids were released from the nucleolar fraction by increasing the ionic strength of the medium to 0.66 suggesting that viroid RNA is present in these subnuclear components in a protein-nucleic acid complex. A purification procedure for nuclei from leaf tissue had to be newly developed; it involves two Percoll density centrifugations as final steps. The nuclei were sonicated and the sonicate fractionated into fractions either highly enriched in nucleoli or in broken chromatin and ribonucleoprotein particles. The viroid content in the different samples was determined by gel electrophoresis. Depending upon the progress of the disease, viroid copy numbers between 200 and 10,000 per cell were observed in homogenized tissue, purified nuclei and in the nucleolar fraction. In chloroplasts, practically no viroids were detected. The results are discussed in the light of current hypotheses about the replication, pathogenicity and origin of viroids. PMID- 11892809 TI - Activation of sarcoplasmic reticular Ca2+ transport ATPase by phosphorylation of an associated phosphatidylinositol. AB - Approximately 1 mol phosphatidylinositol phosphate is formed per mol isolated Ca2+ transport ATPase when the enzyme is incubated with ATP/Mg2+. The phosphorylation of this enzyme-associated phosphatidylinositol represents the alkylphosphate formation described earlier. The phosphatidylinositol phosphate has been found in the hydrophobic core of the enzyme. A complex of phosphatidylinositol phosphate with protein can be extracted with acidic chloroform/methanol. The protein behaves like proteolipid during chromatography on Sephadex LH 60 and binds the radioactively labelled phosphatidylinositol phosphate. The phosphorylation of approximately 1 mol phosphatidylinositol per 100,000 g protein correlates with an enhancement of the Ca2+ transport ATPase activity which is due to an approximately 7-fold enhanced affinity for Ca2+ and an approximately 2-fold enhanced maximal turnover. PMID- 11892811 TI - Structure of D-DNA: 8-fold or 7-fold helix? AB - We have shown that both right- and left-handed uniform helical models (RU and LU models) could be built to give satisfactory agreement with the fibre diffraction data of poly[d(I-C)] in the D-form. Atomic coordinates of these two models are reported in the present work. Molecular transforms of these two models, as well as of the recently published Hoogsteen base-paired 7-fold helical structure of Drew and Dickerson, are given. In view of the work of Drew and Dickerson, attention is drawn to the presence of clear 004 and 008 reflections in the diffraction patterns of poly[d(I-C)] and poly[d(A-T)]. The available data strongly suggest an 8-fold helical structure for the D-form of DNA. PMID- 11892813 TI - Influence of transformation by Rous sarcoma virus on the amount, phosphorylation and enzyme kinetic properties of enolase. AB - Using chicken embryo fibroblasts infected with the NY68 transformation-defective temperature-sensitive mutant of Rous sarcoma virus, the phosphorylation and enzyme kinetic properties of enolase have been studied before, and at different stages after, the onset of transformation. A method for purification of enolase was developed, which minimized dephosphorylation. Two enolase (EC 4.2.1.11) isoenzymes were separated by isoelectric focussing revealing that it was the gammagamma form (pI 5.2-6.7) which had become phosphorylated at tyrosine residues after transformation. The phosphorylation of enolase in tyrosine occurred slowly after shift to the permissive temperature, rising from undetectable levels in phenotypically normal cells, to < 10% of the total phosphoamino acid after 3 h, and reaching 30-50% of the total phosphoamino acid by 16 h. Interestingly, the fraction of phosphorylated enolase molecules declined during transformation from 8% in normal cells to 5% by 16 h after temperature shift, due to a 3- to 5-fold increase in the total amount of enolase present in the transformed cultures. Although transformation had no apparent effect on the K0.5 of enolase (26 +/- 4 microM for 2-phosphoglycerate), its specific activity was reduced by about one third. PMID- 11892812 TI - Growth factor-binding proteases in the murine submaxillary gland: isolation of a cDNA clone. AB - The submaxillary gland of the adult male mouse contains a number of serine proteases, several of which are involved in the proteolytic processing of precursors to growth factors and other biologically active polypeptides. Here we report the isolation and identification of a cDNA clone corresponding to one of the proteases, the type B of the epidermal growth factor-binding protein. A pronounced sequence homology was found between the predicted activation peptide of this protease and the NH2-terminal extension of the nerve growth factor alpha subunit, suggesting that the latter protein has an uncleaved activation peptide attached to its NH2 terminus. PMID- 11892814 TI - Analysis of sequences conferring autonomous replication in baker's yeast. AB - A method is presented for rapid sequencing and mapping of elements which support autonomous replication in yeast. The strategy relies on a novel phage M13 vector which allows detection of ARS (autonomously replicating sequence) function in cloned fragments. Deletion mapping of an ARS element linked to the HO gene of Saccharomyces cerevisiae has identified a 57-bp region 3' to the gene, which is essential for autonomous replication. This region shows sequence homology to other ARS elements. PMID- 11892815 TI - Integration of Ecogpt and SV40 early region sequences into human chromosome 17: a dominant selection system in whole cell and microcell human-mouse hybrids. AB - The dominant selectable gene, Ecogpt, has been introduced, by the calcium phosphate precipitation technique, into normal human fibroblasts, along with the SV40 early region genes. In one transfectant clone, integration of these sequences into human chromosome 17 was demonstrated by the construction of human mouse somatic cell hybrids, selected for by growth in medium containing mycophenolic acid and xanthine. A whole cell hybrid, made between the human transfectant and a mouse L cell, was used as donor of the Ecogpt-carrying human chromosome 17 to 'tribrids' growing in suspension, made by whole cell fusion between a mouse thymoma cell line, and to microcell hybrids made with a mouse teratocarcinoma cell line. Two tribrids contained karyotypically normal human chromosomes 17 and a small number of other human chromosomes, while a third tribrid had a portion of the long arm of chromosome 17 translocated to mouse as its only human genetic material. Two independent microcell hybrids contained a normal chromosome 17 and no other human chromosome on a mouse teratocarcinoma background. These experiments demonstrate the ability to construct human-mouse somatic cell hybrids using a dominant selection system. By applying this approach it should be possible to select for a wide range of different human chromosomes in whole cell and microcell hybrids. In particular, transfer of single human chromosomes to mouse teratocarcinoma cells will allow examination of developmentally regulated human gene sequences after differentiation of such hybrids. PMID- 11892816 TI - Differential regulation of HLA-DR mRNAs and cell surface antigens by interferon. AB - Human interferons-alpha, -beta and -gamma enhance HLA-DR mRNAs in all the human lymphoblastoid and melanoma cell lines studied. The increase concerns both alpha and beta chain mRNAs. Moreover, we show that immune interferon-gamma preferentially enhances class II MHC mRNA. This effect of IFN-gamma on the synthesis of alpha and beta HLA-DR chains has been also analysed by immunoprecipitation. It is abolished by a monoclonal antibody directed against human IFN-gamma. The effect of interferon on the cell surface level of HLA-DR molecules does not always correspond to the enhancement of HLA-DR mRNA. Our experiments suggest that this discrepancy between the enhancement of HLA-DR mRNA and cell surface antigen might be due to a constitutively high level of the corresponding antigens on several of the human cells studied. PMID- 11892817 TI - Somatic variants of the level of expression of a cell surface antigen. AB - Somatic variants with constitutive changes in the expression of the cortical thymocyte differentiation antigen HTA 1 were derived from the T-cell leukemia line Molt 4 using monoclonal antibody NA1/34 and the fluorescent activated cell sorter. Cells with the highest and lowest fluorescence were sorted and expanded. After several cycles, high expressor variants, with 10- to 12-fold increased surface HTA 1 and low expressors, with a level of one third relative to the wild type, were obtained. The stability of the high expressors was improved by cloning. In spite of the large differences in the level of HTA 1 between the various mutants and the wild-type, the expression of HLA-A,B,C or its increase following interferon-alpha stimulation, remained unchanged. Although in HTA 1 there was only a trace of beta2-microglobulin (beta2m) detected by lactoperoxidase labelling of the high expressor variants, significant levels of both beta2m and HTA 1 light chain beta(t) were observed in gels stained with Coomassie blue. The physiological association of beta(t) with HTA 1 was confirmed by the substantial increase of beta(t) which parallels the increase of HTA 1. PMID- 11892818 TI - The opine synthase genes carried by Ti plasmids contain all signals necessary for expression in plants. AB - Signals necessary for in vivo expression of Ti plasmid T-DNA-encoded octopine and nopaline synthase genes were studied in crown gall tumors by constructing mutated genes carrying various lengths of sequences upstream of the 5' initiation site of their mRNAs. Deletions upstream of position -294 did not interfere with expression of the octopine synthase gene while those extending upstream of position -170 greatly reduced the gene expression. The estimated size of the octopine synthase promoter is therefore 295 bp. The maximal length of 5' upstream sequences involved in the in vivo expression of the nopaline synthase gene is 261 bp. Our results also demonstrated that Ti plasmid-derived sequences contain all signals essential for expression of opine synthase genes in plants. Expression of these genes, therefore, is independent of the direct vicinity of the plant DNA sequences and is not activated by formation of plant DNA and T-DNA border junction. PMID- 11892819 TI - Transcription from the SV40 early-early and late-early overlapping promoters in the absence of DNA replication. AB - Transcription for a hybrid SV40 promoter-beta globin coding sequence recombinant initiates from both early-early (EE) and late-early (LE) SV40 start sites (EES and LES) in the absence of DNA replication. The 72-bp repeat is essential to potentiate the elements of the two overlapping EE and LE promoters (EEP and LEP). Two current models, which can account for the EE to LE shift in RNA chain initiation during the SV40 replication cycle, are that LE transcription is linked to replication and occurs on newly replicated DNA molecules or that there are two promoter elements, a stronger EEP and a weaker LEP, T antigen repressing the EEP late in infection. Our results support the second model. A 5'-TATTTAT-3' to 5' TATCGAT-3' mutation in the putative SV40 TATA box decreases transcription from EES, increases transcription from LES, and inhibits DNA replication. Therefore, this element acts as a classical TATA box for transcription, and yet is also important for DNA replication. PMID- 11892821 TI - Adverse events in the 'medical' ward. PMID- 11892820 TI - Cytoplasmic regulation of 5S RNA genes in nuclear-transplant embryos. AB - In the normal development of Xenopus laevis, genes for oocyte-type and somatic type 5S RNAs are both expressed in late blastulae. Estimates of rates of synthesis indicate that the oocyte-type genes (5Sooc) undergo at least a 100-fold reduction in transcriptional activity between the end of oogenesis and the late blastula stage, and at least a further 20-fold reduction during gastrulation. When neurula nuclei, with inactive 5Sooc genes, were transplanted to enucleated eggs, the resulting nuclear-transplant embryos showed a reactivation of 5Sooc genes in blastulae and a subsequent inactivation after this stage. This effect is not explicable by a differential stability of the two types of 5S RNA. We conclude that egg or early embryo cytoplasm must contain components which can continuously regulate 5S gene expression, and that the mechanism by which 5Sooc genes are developmentally inactivated does not persist through mitosis in early embryos. These results have been obtained by a new procedure in which oocyte- and somatic-type 5S RNAs are separated in a 4 M urea-15% acrylamide gel. PMID- 11892822 TI - The benefits of using clinical pathways for managing acute paediatric illness in an emergency department. AB - The aim of this study was to provide an evaluation of the overall effectiveness of using a number of clinical pathways in treating common acute paediatric conditions in an emergency department. This was a before and after study conducted on the effectiveness of three clinical pathways (gastroenteritis, asthma, and croup) in the emergency department of the Children's Hospital at Westmead, conducted over two separate yearly periods January to December 1996 and January to December 1999 representing before and after the introduction of clinical pathways in the emergency department. The main outcomes of the effectiveness of the pathways, namely admission to an in-patient bed, length of hospital stay and re-presentation after discharge from the ED were compared. Other outcomes of interest such as parental satisfaction and patient waiting times were also presented. Any deviation from a key clinical pathway process was reported. A total of 2854 children were managed by a clinical pathway compared to 2680 children managed before clinical pathways were introduced. The admission rate was reduced by threefold (9.1% compared to 23.6%) with a twofold reduction in length of hospital stay (32.7 h compared to 17.5 h). In 3.6% of children using a clinical pathway an unscheduled medical visit or re-presentation to the emergency department occurred after discharge, compared to 4.9% before the use of clinical pathways. No adverse events were reported in these children. In 76 cases deviation from a clinical pathway process was reported. High parental satisfaction was reported for clinical pathways throughout the study. Clinical pathways in this emergency department allowed rapid stabilisation of children, reducing admission rate, with a shortened length of hospital stay and few patients re-presenting after discharge and were well accepted by parents. PMID- 11892823 TI - Hospital stay and discharge outcomes after knee arthroplasty. AB - The impact of shorter hospital lengths of stay on patient outcomes at discharge from acute care after knee arthroplasty was investigated in a prospective observational outcome study at three Melbourne public hospitals during a 5-month period from October 1999 to March 2000. The participants were 105 consecutive patients (35 at each hospital), with a mean age of 71 years. Outcome measures were length of stay, destination (home or rehabilitation) and functional mobility at discharge from the acute care facility. During the study period mean hospital length of stay across the three hospitals was 6.5 days, more than 30% less than the Victorian average for the preceding year. This was associated with high rates of discharge to rehabilitation facilities (mean 64%), with rates varying between the three hospitals (97%, 57% and 40%). However, in each hospital, one-third of this group had already achieved a level of independent functional mobility adequate for discharge home, highlighting an apparent influence of non-clinical factors on discharge decisions, including pressure to decrease length of stay, hospital policy and availability of a rehabilitation bed. Ways of achieving discharge directly home for a greater number of patients following knee arthroplasty and of determining optimal length of stay are discussed. PMID- 11892825 TI - An evaluation of outcome after severe trauma. AB - There is a perception that patients with severe trauma either die soon after the trauma or survive. This study evaluated 123 patients with severe trauma until 6 months after discharge from the Intensive Care Unit (ICU). At baseline, the median Injury Severity Score was 29 (interquartile range 20-38) and the median Acute Physiology and Chronic Health Evaluation II score was 13 (interquartile range 9-15). Injuries related to road transport accounted for 79% (97/123) of the injuries and the patients were predominantly male (66%) and young (75% < 38 years of age). The median stay in ICU was 11 days. Ten per cent of the patients (12/123) died during the study period, but one-quarter of the deaths were in hospital after discharge from ICU and one-quarter of the deaths occurred after discharge from hospital. It is impossible to carefully document the mortality due to severe trauma unless the survivors of ICU are reviewed after discharge from hospital. PMID- 11892824 TI - A prospective, physician self-reported adverse incident audit on a general medical unit. AB - There is some evidence that physician self-reporting is an efficient and effective way of collecting data on adverse incidents in health care. This study tested a simple prospective adverse incident audit, self-reported by physicians, on a general medical unit. A total of 158 reports were collected over a 6-month period covering a wide range of quality issues, including, but not limited to, safety issues. One-third of reported incidents occurred within 48 h of hospitalization. One-half of incidents were associated with harm or inconvenience to patients. Reported incidents fell into easily classifiable groups, and the data was used as a platform for a coordinated approach to quality improvement within the department. It is concluded that this technique is an easily implementable addition to the more traditional methods used for quality improvement within general medicine. PMID- 11892826 TI - A quality assurance activity to improve discharge communication with general practice. AB - The aim of this study was to describe and evaluate a method of improving the completion of discharge summary information. As a quality assurance activity, a self-carbonating form for both the patient record and discharge letter was introduced to a general practice casualty (GPC) in Sydney, Australia, involving approximately 40 community GPs. Evaluation involved assessment of the change associated with this intervention on a cohort of GPs using logistic regression analysis, clustering at the level of the treating GP. The main outcome measure was completion of a discharge letter to the patient's usual GP. Provision of discharge letters increased from 30% prior to the intervention to 90% following it (P< 0.0005; OR 11.9; 95% CI 8.6-16.4). It was concluded that solutions to problems of quality are best developed at the system level. The use of a self carbonating form to simultaneously generate a discharge letter and the medical record might be used by other similar services with good effect. PMID- 11892827 TI - Use of clinical audit for revalidation: is it sufficiently accurate? AB - In order to provide better patient care, clinicians will be subject to revalidation and re-certification. This may be partially based on existing and ongoing data collection, yet many units fail to incorporate mechanisms that validate the data that may be used. The accuracy of audit data was evaluated in a unit that has been using commercially available audit software for over 10 years. A total of 655 consecutive surgical admissions were documented over a 6-month period and errors in data collection and entry were gathered and analyzed. An overall accuracy of 90.5% was confirmed but examination of the data found them to be open to misinterpretation. Moreover, 13% of errors were made during a single week when locum staff were involved. The study highlights the fallibility of data collection during audit, and urges caution if using such data when judging performance-related issues as part of the process of appraisal. PMID- 11892828 TI - Ensuring accuracy of clinical data is only part of the audit process. PMID- 11892829 TI - Eradicating Helicobacter pylori in patients with a past history of peptic ulcer: is the juice worth the squeeze? AB - The efficacy of Helicobacter pylori eradication for H. pylori associated duodenal ulcer disease is beyond dispute. However, little attention has been paid to how feasible it is in primary care to undertake H. pylori eradication for patients with a past history of peptic ulcer disease. Patients identified by computer search of three general practices with a documented history of peptic ulcer disease were invited to attend for H. pylori testing and eradication therapy if H. pylori positive. A total of 101 patients were identified from a combined practice size of 24,780 of whom 34 were eligible for testing. Twenty-one per cent (seven patients) declined testing, 3% (one patient) refused therapy and another 21% (seven patients) were H. pylori negative. Helicobacter pylori eradication was highly successful (95%) and had an effect on prescription and reconsultation rates at 12 months. Eradicating H. pylori in patients with a past history of peptic ulcer disease not associated with non-steroidal anti-inflammatory drug ingestion is a challenging goal in general practice. PMID- 11892830 TI - Quality management and the Emergency Services Enhancement Program. AB - Since the introduction of the Emergency Services Enhancement Program (ESEP) in Victoria in 1995, improvements have been demonstrated in the indicators relating to Emergency waiting times, ambulance bypass rates and inpatient bed access block. This study focuses on staff perceptions of changes in these indicators, factors perceived to influence performance improvements and the extent to which ESEP is perceived to have contributed to overall patient care. A questionnaire was directed at four focus groups within each of the hospitals participating in ESEP. These were Chief Executive Officers, Emergency Department Directors and Nurse Unit Managers, bed coordinators and personnel from the Emergency Department floor. A total of 101 staff responded. Emergency Department staff were generally accurate in their perceptions of performance changes. The most important factors effecting the changes were perceived to be changes in staff profile, management of patient flow through the department, changes in administrative policies and changes in work practices. Staff perceived that patient care has improved by 10% since 1995 and that ESEP has contributed 8% of this improvement. Staff have perceived improvements in ESEP performance indicators consistent with actual changes. The possible mechanisms by which these changes have occurred are presented and discussed. Factor analysis indicated that changes perceived to be most likely to result in improvements were: changes in staff profile (seniority), managing the flow of patients through emergency departments, changing administrative policies, changes in work practices and changes in staff numbers. Improvements in patient care were considered partly due to ESEP. In addition, ESEP has raised awareness of quality management issues. PMID- 11892831 TI - Rational prescribing for childhood pneumonia. AB - The aim of this study was to assess the management and prescribing patterns for community acquired pneumonia in children in a provincial hospital setting and, further, to discover the evidence on which treatment choices were made and ascertain the need for management guidelines. The method employed was to obtain the relevant data by a retrospective audit of the case notes of children admitted with pneumonia to Wairau Hospital, Blenheim, New Zealand. The findings indicated that there were 12 different treatment regimens employed. There appeared to be no rational basis for the choices made and no microbiological evidence to support the frequent use of broad spectrum antibiotics. No difference in clinical efficacy was found between the main regimens used. It is concluded that a simple management protocol for childhood pneumonia using a narrow spectrum antibiotic initially would be as effective, more logical and cheaper. PMID- 11892832 TI - Aspects of the Pharmaceuticals Benefits Scheme. PMID- 11892833 TI - Phenomenon of quality and health-care. PMID- 11892834 TI - Insulators coupled to a minimal bidirectional tet cassette for tight regulation of rAAV-mediated gene transfer in the mammalian brain. AB - Recombinant AAV is increasingly becoming the vector of choice for many gene therapy applications in the CNS, due to its lack of toxicity and high level of sustained expression. With recent improvements in the generation of pure, high titer vector stocks, the regulation of gene expression is now a key issue for successful translation of gene therapy-based treatments to the clinic. The level of the transgene protein may need to be maintained within a narrow therapeutic window for the successful treatment of human disease. The doxycycline responsive system directs a dose-responsive, tightly regulated level of gene expression and has been used successfully in transgenic mouse models. Here, we have optimized an autoregulatory, bidirectional doxycyline responsive cassette specifically for use in rAAV. We minimized the size of the cassette and decreased the basal leakiness of the system, leading to tight regulation in the rat PMID- 11892835 TI - Dominant negative c-jun gene transfer inhibits vascular smooth muscle cell proliferation and neointimal hyperplasia in rats. AB - We previously reported that activator protein-1 (AP-1), containing c-Jun, is rapidly activated in balloon-injured artery. Therefore, we examined the role of c Jun in vascular smooth muscle cell (SMC) proliferation, by using in vitro and in vivo gene transfer techniques. (1) Serum (2%) stimulation significantly increased AP-1 DNA binding activity in aortic SMCs, followed by the increase in both 3H thymidine incorporation and cell number. Aortic SMCs were infected with recombinant adenovirus containing TAM67, a dominant negative c-Jun lacking transactivation domain of wild c-Jun (Ad-DN-c-Jun), to specifically inhibit AP-1. Ad-DN-c-Jun significantly inhibited serum-induced SMC proliferation, by inhibiting the entrance of SMC into S phase. (2) The effect of DN-c-Jun was examined on balloon injury-induced intimal hyperplasia in rats. Before balloon injury, DN-c-Jun was transfected into rat carotid artery using the hemagglutinating virus of Japan-liposome method. In vivo transfection of DN-c-Jun significantly inhibited vascular SMC proliferation in the intima and the media and subsequently prevented intimal thickening at 14 days after balloon injury. We obtained the first evidence that DN-c-Jun gene transfer prevented vascular SMC proliferation in vitro and in vivo, and c-Jun was involved in balloon injury induced intimal hyperplasia. Thus, AP-1 seems to be the new therapeutic target for treatment of vascular diseases. PMID- 11892836 TI - Recombinant adeno-associated virus vector-transduced vascular endothelial cells express the thrombomodulin transgene under the regulation of enhanced plasminogen activator inhibitor-1 promoter. AB - We were able to facilitate plasminogen activator inhibitor 1 (PAI-1) promoter activity approximately by 14-fold using an enhancer element. This enhanced PAI-1 promoter has a strong basal activity, comparable to CAG promoter activity, and has a response similar to the PAI-1 promoter with respect to TGFbeta 1 and TNFalpha stimulation. The characteristics of the enhanced PAI-1 promoter are thought to be suited to timely and tissue-specific expression of anticoagulant molecules in the vascular cells. Thus, we developed recombinant adeno-associated virus (rAAV) vectors using the enhanced PAI-1 promoter and were successful in transducing vascular endothelial cells to express the thrombomodulin transgene under the regulation of the enhanced PAI-1 promoter in vitro. Thromobomodulin transgene expression driven by the enhanced PAI-1 promoter in rAAV vector transduced cultured endothelial cells was between 600- and 1000-fold higher than constitutive thrombomodulin gene expression in cultured human umbilical vein endothelial cells and was up-regulated by TGFbeta1 and TNFalpha stimulation which may down-regulate endogenous thrombomodulin gene expression in endothelial cells. The brain vascular endothelial cells of Mongolian gerbils could also be transduced by the same rAAV vector in vivo. Transduction of endothelial cells by rAAV vectors to express enhanced PAI-1 promoter-driven transgenes may be a useful gene therapy approach for vascular diseases. PMID- 11892837 TI - Therapy of lung metastases through combined vaccination with carcinoma cells engineered to release IL-13 and IFN-gamma. AB - TS/A spontaneous mouse mammary adenocarcinoma cells were engineered to release interferon-gamma (IFN-gamma), a Th1 cytokine (TS/A-IFNgamma) and interleukin-13 (IL-13), a Th2 cytokine (TS/A-IL13). Mice bearing lung micrometastases induced by parental TS/A cells received repeated subcutaneous vaccinations with TS/A-IFN gamma admixed with TS/A-IL13 engineered cells. This combined treatment cured up to 75% of mice, whereas vaccinations with either TS/A-IFNgamma or TS/A-IL13 alone cured only 20-40% of mice. Combined TS/A-IL13 and TS/A-IFNgamma therapeutic vaccinations elicited a reactive infiltrate of CD4+ and CD8+ lymphocytes in lung metastases and an increased production of IFN-gamma in the spleen and lung, suggesting a shift of the immune response toward the Th1 type. The type of infiltrating cells along with the lack of efficacy in T cell-deficient mice point to a major role of T cells. In conclusion, no antagonism but a synergistic and effective definitive cure stems from the combined vaccination with tumor cells engineered to release a Th1 and a Th2 cytokine. PMID- 11892838 TI - Cell cycle arrest is sufficient for p53-mediated tumor regression. AB - p53 gene therapy can induce tumor regression, but the low efficacy of in vivo gene transfer has greatly hampered the mechanistic analysis of this antitumoral activity. We therefore used a p53-null human NSCLC cell line in which we reintroduced the wild-type p53 gene under control of a tetracycline-dependent promoter. P53 induction provokes cell cycle arrest in G0/G1 and G2/M phase, an up regulation of p21, a down-regulation of cyclin B1 and appearance of senescence features without down-regulation of human telomerase reverse transcriptase. No detectable morphological changes of apoptosis nor procaspase-3 activation are observed. In subcutaneous tumors grafted in nude mice, the induction of p53 expression leads to a complete and longlasting tumor regression in 28 days which is associated with cell cycle arrest, but not detectable apoptosis nor inhibition of angiogenesis. These results show that irreversible cell cycle arrest is sufficient to elicit tumor regression after p53 gene transfer in p53-deficient tumor cells. PMID- 11892839 TI - Degenerated pIX-IVa2 adenoviral vector sequences lowers reacquisition of the E1 genes during virus amplification in 293 cells. AB - A critical issue for E1-deleted adenoviral vectors manufactured from 293 cells is the emergence of replication-competent adenovirus (RCA). These contaminants arise through homologous recombination between identical sequences framing the E1 locus displayed by 293 cells, and the vector backbones. Modified recombinogenic sequences (syngen) were thus introduced within the vector backbone, and virus viability and RCA emergence were assessed. Syngen#1 is a synthetic sequence displaying silent point mutations in the pIX and IVa2 coding regions. A side by side comparison of Ad5CMV/p53 (E1-deleted adenovirus expressing the p53 tumor suppressor gene) and AVdeltaE1#1CMV/p53 (with syngen#1 in place of wild-type sequences) demonstrated a normal productivity for the modified construct. The altered sequences did not impair p53-mediated apoptosis in a model tumor cell line. Most importantly, a statistically significant decrease in terms of RCA occurrence could also be demonstrated. Degenerescence of the recombinogenic sequences could be further accentuated by modifying noncoding pIX region (syngen #2), with no effect on virus productivity and stability. We concluded that these vector modifications constitute a feasible strategy to reduce RCA emergence during amplification in 293 cells. This approach could also be applied to decrease reincorporation of the E1 genes during amplification of deltaE1deltaE4 vectors in 293/E4-trans-complementing cells. PMID- 11892840 TI - Bypassing tumor-specific and bispecific antibodies: triggering of antitumor immunity by expression of anti-FcgammaR scFv on cancer cell surface. AB - We have developed a novel immunostimulatory molecule against tumor cells, composed of an anti-FcgammaRIII (CD16) scFv fused to the platelet-derived growth factor receptor (PDGFR) transmembrane region. This fusion molecule was stably expressed on the tumor cell surface and retained the ability of the parental antibody to bind soluble CD16. Tumor cells expressing anti-CD16 scFv triggered the release of IL-2 by Jurkat-CD 16/gamma cells and of TNFalpha by monocytes when co-cultured with these cells. Furthermore, NK cells could kill scFv-transfected HLA+ class I H1299 lung carcinoma tumor cells, but not the parental cells, indicating that anti-CD16 scFv tumor expression prevents the killer inhibitory receptor (KIR)-mediated inhibition of NK cell cytotoxicity. This anti-CD16 scFv tumor expression also enhanced tumor phagocytosis by IFNgamma-activated macrophages, a mechanism known to induce a protective long-term adaptative immunity to tumors. In vivo Winn tests performed in SCID mice showed that the expression of anti-CD16 scFv on tumor cells, but not of the negative control anti phOx scFv, prevented tumor cell growth. Thus, expression of FcR antibodies or other FcR-specific ligands on tumor cells represents a novel and potent antibody based gene therapy approach, which may have clinical applications in cancer PMID- 11892841 TI - Development of a Langerhans cell-targeted gene therapy format using a dendritic cell-specific promoter. AB - Langerhans cells (LC), which are a skin-specific member of the dendritic cell (DC) family of antigen presenting cells, play critical roles in the initiation of cellular immune responses in the skin. We developed a LC-targeted gene therapy format in this study, aimed at the establishment of in situ protocols for genetic manipulation of LC function. Dectin-2 is a unique C-type lectin that is expressed selectively by DC, including epidermal LC. A 3.2 kb 5' flanking fragment isolated from the mouse dectin-2 gene, termed the dectin-2 promoter (pDec2), exhibited significant transcriptional activities in epidermal-derived DC lines of the XS series, but not in any of the tested non-DC lines. When pDec2-driven luciferase gene (pDec2-Luc) or enhanced green fluorescence protein gene (pDec2-EGFP) was delivered to mouse skin using the gene gun, expression of the corresponding gene product was observed in the epidermal compartment almost exclusively by the IA+ population (ie LC). LC in the gene gun-treated sites showed features of mature DC and they migrated to the draining lymph node, suggesting that LC-targeted gene expression may lead to the development of immune responses. In fact, EGFP specific cellular immune responses became detectable after gene gun-mediated delivery of pDec2-EGFP plasmid. These results introduce a new concept that LC function can be genetically manipulated in situ by the combination of gene gun mediated DNA delivery and a DC-specific promoter. PMID- 11892843 TI - A highly efficient and consistent method for harvesting large volumes of high titre lentiviral vectors. AB - Lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein (VSV G) are emerging as the vectors of choice for in vitro and in vivo gene therapy studies. However, the current method for harvesting lentivectors relies upon ultracentrifugation at 50,000 g for 2 h. At this ultra-high speed, rotors currently in use generally have small volume capacity. Therefore, preparations of large volumes of high-titre vectors are time-consuming and laborious to perform. In the present study, viral vector supernatant harvests from vector-producing cells (VPCs) were pre-treated with various amounts of poly-L-lysine (PLL) and concentrated by low speed centrifugation. Optimal conditions were established when 0.005% of PLL (w/v) was added to vector supernatant harvests, followed by incubation for 30 min and centrifugation at 10,000 g for 2 h at 4 degrees C. Direct comparison with ultracentrifugation demonstrated that the new method consistently produced larger volumes (6 ml) of high-titre viral vector at 1 x 10(8) transduction unit (TU)/ml (from about 3,000 ml of supernatant) in one round of concentration. Electron microscopic analysis showed that PLL/viral vector formed complexes, which probably facilitated easy precipitation at low-speed concentration (10,000 g), a speed which does not usually precipitate viral particles efficiently. Transfection of several cell lines in vitro and transduction in vivo in the liver with the lentivector/PLL complexes demonstrated efficient gene transfer without any significant signs of toxicity. These results suggest that the new method provides a convenient means for harvesting large volumes of high-titre lentivectors, facilitate gene therapy experiments in large animal or human gene therapy trials, in which large amounts of lentiviral vectors are a prerequisite. PMID- 11892842 TI - Induction of molecular chimerism by gene therapy prevents antibody-mediated heart transplant rejection. AB - In order for xenotransplantation to become a clinical reality, and fulfill its promise of overcoming shortages of human organs and tissues, rejection mediated by the host's immune system must first be overcome. In primates, preformed natural antibodies that bind the carbohydrate antigen Galalpha1-3Galbeta1-4GIcNAc R (alphaGal), which is synthesized by UDP galactose:beta-D-galactosyl-1,4-N acetyl-D-glucosaminide alpha(1-3)galactosyltransferase (E.C. 2.4.1.151) or simply alphaGT, mediate rigorous rejection of transplanted pig organs and tissues. In alphaGT knockout mice (GT0 mice), which like humans contain in their serum antibodies that bind alphaGal, expression of a retrovirally transduced alphaGT in bone marrow-derived cells is sufficient to prevent production of alphaGal reactive antibodies. Here, we demonstrate that reconstitution of lethally irradiated GT0 mice with alphaGT-transduced bone marrow cells from GT0 littermates prevents antibody-mediated rejection of cardiac transplants from wild type mice. These data suggest that gene therapy can be used to induce immunological tolerance to defined antigens and thereby overcome transplant rejection. PMID- 11892844 TI - Antimicrobial peptides from the skin of the Japanese mountain brown frog, Rana ornativentris. AB - Six peptides with antimicrobial activity were isolated from an extract of freeze dried skin of the Japanese mountain brown frog Rana ornativentris. Two structurally related peptides (brevinin-20a GLFNVFKGALKTAGKHVAGSLLNQLKCKVSGGC, 11 nmol/g dried tissue, and brevinin-20b GIFNVFKGALKTAGKHVAGSLLNQLKCKVSGEC, 170 nmol/g) belong to the brevinin-2 family, previously identified in Asian and European, but not North American, Ranid frogs. Four peptides (temporin-10a FLPLLASLFSRLL.NH2, 13 nmol/g; temporin-10b FLPLIGKILGTI L.NH2, 350 nmol/g; temporin-10c FLPLLASLFSRLF.NH2, 14 nmol/g; and temporin-10d FLPLLASLFSGLF.NH2, 8 nmol/g) are members of the temporin family first identified in the European common frog Rana temporaria but also found in the skins of North American Ranids. The brevinin-2 peptides showed broad-spectrum activity against the gram-positive bacterium, Staphylococcus aureus, the gram-negative bacterium, Escherichia coli and the yeast Candida albicans, whereas the temporins showed potent activity only against S. aureus. The brevinins and temporins belong to the class of cationic antimicrobial peptides that adopt an amphipathic alpha-helical conformation but it is significant that temporin-10d, which lacks a basic amino acid residue, is still active against S. aureus (minimum inhibitory concentration=13 microM compared with 2 microM for temporin-10a). This suggests that strong electrostatic interaction between the peptide and the negatively charged phospholipids of the cell membrane is not an absolute prerequisite for antimicrobial activity. PMID- 11892845 TI - Engineering the prion protein using chemical synthesis. AB - In recent years, the technology of solid-phase peptide synthesis (SPPS) has improved to the extent that chemical synthesis of small proteins may be a viable complementary strategy to recombinant expression. We have prepared several modified and wild-type prion protein (PrP) polypeptides, of up to 112 residues, that demonstrate the flexibility of a chemical approach to protein synthesis. The principal event in prion disease is the conformational change of the normal, alpha-helical cellular protein (PrPc) into a beta-sheet-rich pathogenic isoform (PrP(Sc)). The ability to form PrP(Sc) in transgenic mice is retained by a 106 residue 'mini-prion' (PrP106), with the deletions 23-88 and 141-176. Synthetic PrP106 (sPrP106) and a His-tagged analog (sPrP106HT) have been prepared successfully using a highly optimized Fmoc chemical methodology involving DCC/HOBt activation and an efficient capping procedure with N-(2 chlorobenzyloxycarbonyloxy) succinimide. A single reversed-phase purification step gave homogeneous protein, in excellent yield. With respect to its conformational and aggregational properties and its response to proteinase digestion, sPrP106 was indistinguishable from its recombinant analog (rPrP106). Certain sequences that proved to be more difficult to synthesize using the Fmoc approach, such as bovine (Bo) PrP(90-200), were successfully prepared using a combination of the highly activated coupling reagent HATU and t-Boc chemistry. To mimic the glycosylphosphatidyl inositol (GPI) anchor and target sPrP to cholesterol-rich domains on the cell surface, where the conversion of PrPc is believed to occur, a lipophilic group or biotin, was added to an orthogonally side-chain-protected Lys residue at the C-terminus of sPrP sequences. These groups enabled sPrP to be immobilized on either the cell surface or a streptavidin-coated ELISA plate, respectively, in an orientation analogous to that of membrane-bound, GPI-anchored PrPc. The chemical manipulation of such biologically relevant forms of PrP by the introduction of point mutations or groups that mimic post-translational modifications should enhance our understanding of the processes that cause prion diseases and may lead to the chemical synthesis of an infectious agent. PMID- 11892846 TI - Spermidine as a potential biosynthetic precursor to the 1,5 diazabicyclo[4:3:o]nonene residue in the efrapeptins. AB - Efrapeptins are a group of microheterogeneous polypeptide antibiotics produced by the fungus Tolypocladium niveum, which are potent inhibitors of mitochondrial F1 ATPase. Efrapeptins contain an unusual 1,5-diazabicyclo[4:3:0]nonene (DBN) residue at the C-terminus. This study is driven by the hypothesis that the DBN residue could, in principle, arise by oxidative cyclization of a spermidine moiety. Electrospray ionization mass spectrometry of the peptide antibiotics 'elvapeptins' from T niveum establishes the presence of a C-terminal spermidine residue. Conversion of elvapeptins to efrapeptins by CuCl/pyridine demonstrates the transformation of the spermidine residue to the 1,5-diazabicyclo[4:3:0]nonene system by oxidative cyclization. PMID- 11892847 TI - An in vivo study of novel bioactive peptides that inhibit the growth of Escherichia coli. AB - We have created a system in which synthetically produced novel bioactive peptides can be expressed in vivo in Escherichia coli. Twenty thousand of these peptides were screened and 21 inhibitors were found that could inhibit the growth of E. coli on minimal media. The inhibitors could be placed into one of two groups, 1 day inhibitors, which were partially inhibitory, and 2-day inhibitors, which were completely inhibitory. Sequence analysis showed that two of the most potent inhibitors were actually peptide-protein chimeras in which the peptides had become fused to the 63 amino acid Rop protein which was also contained in the expression vector used in this study. Given that Rop is known to form an incredibly stable structure, it could be serving as a stabilizing motif for these peptides. Sequence analysis of the predicted coding regions from the next 10 most inhibitory peptides showed that four of the 10 peptides contained one or more proline residues either at or very near the C-terminal end of the peptide which could act to prevent degradation by peptidases. Collectively, based on what we observed in our screen of synthetic bioactive peptides that could prevent the growth of E. coli and what has been learned from structural studies of naturally occurring bioactive peptides, the presence of a stabilizing motif seems to be important for small peptides, if they are to be biologically active. PMID- 11892849 TI - Conformational comparison of mu-selective endomorphin-2 with its C-terminal free acid in DMSO solution, by 1H NMR spectroscopy and molecular modeling calculation. AB - In order to make clear the structural role of the C-terminal amide group of endomorphin-2 (EM2, H-Tyr-Pro-Phe-Phe-NH2), an endogenous mu-receptor ligand, in the biological function, the solution conformations of endomorphin-2 and its C terminal free acid (EM2OH, H-Tyr-Pro-Phe-Phe-OH), studied using two-dimensional 1H NMR measurements and molecular modeling calculations, were compared. Both peptides were in equilibrium between the cis and trans isomers around the Tyr-Pro omega bond in a population ratio of approximately/= 1:2. The lack of significant temperature and concentration dependence of NH protons suggested that the NMR spectra reflected the conformational features of the respective molecules themselves. Fifty possible 3D structures for the each isomer were generated by the dynamical simulated annealing method under the proton-proton distance constraints derived from the ROE cross-peaks. These energy-minimized conformers, which were all in the phi torsion angles estimated from J(NHCalphaH) coupling constants within +/- 30 degrees, were then classified in groups one or two according to the folding backbone structures. All trans and cis EM2 conformers adopt an open conformation in which their extended backbone structures are twisted at the Pro2-Phe3 moiety. In contrast, the trans and cis conformers of EM2OH show conformational variation between the 'bow'-shaped extended and folded backbone structures, although the cis conformers of its zwitterionic form are refined into the folded structure of the close disposition of C- and N-terminal groups. These results indicate clearly that the substitution of carboxyl group for C-terminal amide group makes the peptide flexible. The conformational requirement for mu-receptor activation has been discussed based on the active form proposed for endomorphin-1 and by comparing conformational features of EM2 and EM2OH. PMID- 11892848 TI - Solution structures of the N-terminal domain of histone H4. AB - Histones, nuclear proteins that interact with DNA to form nucleosomes, are essential for both the regulation of transcription and the packaging of DNA within chromosomes. The N-terminal domain of histone H4 contains four acetylation sites at lysine residues and may play a separate role in chromatin structure from the remainder of the H4 chain. We performed circular dichroism and NMR characterization of both native (H4NTP) and acetylated (Ace-H4NTP) peptides containing N-terminal acetylation domain of histone H4 for various pH environments. Data from CD and NMR suggested that H4NTP exhibited a pH-dependent conformational change, whereas the Ace-H4NTP is insensitive to pH change. However, both peptides showed a defined structural form at acidic pH environments. The solution structure for Ace-H4NTP shows two structurally independent regions comprising residues of Leu10-Gly13 and Arg19-Leu22, demonstrating relatively well-defined turn-type structures. Our results suggest that N-terminal acetylated region of H4 prefers an extended backbone conformation at neutral pH, however, upon acetylation, the regions containing lysine residues induce structural transition, having defined structural form for its optimum function. PMID- 11892850 TI - Helix-stabilizing effects of the pentapeptide KIFMK and its related peptides on the sodium channel inactivation gate peptides. AB - We have previously found by NMR and CD spectroscopic studies that the helical content of the sodium channel inactivation gate-related peptide (Ac-GGQDIFMTEEQK NH2; MP-1A) in 80% trifluoroethanol solutions was increased by adding a pentapeptide, KIFMK. In order to study in further detail whether the presence of the IFM motif and the two lysine residues is a prerequisite for stabilizing the helical conformation, we examined interactions between various oligopeptides (RIFMR, KIFMTK, KIQMK, KAFAK, KIIIK) and MP-1A and its related peptides; that is, MP-2A in which Phe was replaced by Gln, MP-1MMA in which Thr was replaced by Met, MP-1TA in which Thr was removed from MP-1A, and MP-1A' in which L-Phe was replaced by D-Phe. It was found that the IFM motif was absolutely necessary in both the oligopeptide and the inactivation gate peptide. This finding means that hydrophobic interactions are operative between KIFMK and MP-1A. In contrast, KIFMK destabilized the helical structure of MP-1MMA, MP-1TA, and MP-1A', showing that the conformation around the IFM motif in the inactivation gate peptides is an important factor. It was concluded that the IFM motif and the two Lys residues are a prerequisite for effectively stabilizing the alpha-helix of MP-1A. PMID- 11892851 TI - Solid-phase synthesis of peptides containing the spin-labeled 2,2,6,6 tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC). AB - 2,2,6,6-Tetramethylpiperidine-1-oxyl-4-amino4-carboxylic acid (TOAC) is a nitroxide spin-labeled, achiral Calpha-tetrasubstituted amino acid recently shown to be not only an effective beta-turn and 3(10)/alpha-helix promoter in peptides, but also an excellent rigid electron paramagnetic resonance probe and fluorescence quencher. Here, we demonstrate that TOAC can be effectively incorporated into internal positions of peptide sequences using Fmoc chemistry and solid-phase synthesis in an automated apparatus. PMID- 11892852 TI - Structure-function studies on the amphibian peptide brevinin 1E: translocating the cationic segment from the C-terminal end to a central position favors selective antibacterial activity. AB - Brevinin 1E, which has the sequence FLPLLAGLAANFLPKIFCKITRKC, is an antimicrobial peptide isolated from the skin secretions of the European frog Rana esculenta. Both the linear and the disulfide-bridged forms have relatively broad-spectrum antibacterial as well as hemolytic activities. The antibacterial and hemolytic activities and biophysical properties of synthetic peptides corresponding to brevinin 1E and its analog in which the segment CKITRKC has been transposed to a central location resulting in the sequence FLPLLAGLCKITRKCAANFLPKIF have been investigated. Our studies indicate that the analog peptide has antibacterial activity comparable with brevinin 1E, but with considerably reduced hemolytic activity. The linear variant of the analog has no hemolytic activity, unlike the linear form of brevinin 1E. The biological activities can be explained on the basis of relative affinities for anionic and zwitterionic lipids. A cluster of cationic amino acids flanked on one side by a hydrophobic stretch of amino acids and another side composed of apolar amino acids appears to favor preferential antibacterial activity. PMID- 11892853 TI - Parental influence on sibling caregiving for people with severe mental illness. AB - Adult siblings of people with serious mental illness are increasingly being called upon to serve as caregivers for their loved ones. The present study investigated 111 adults' reports of their relationships with their afflicted siblings and with their parents in an attempt to explain well siblings' reports of: (1) current caregiving, (2) hypothetical caregiving willingness, and (3) future intention to care for their brother or sister with mental illness. Results of hierarchical multiple regression analyses indicate that perceived sibling need, sibling affection, reciprocity with ill siblings, felt obligation toward parents, and parental requests for help with caregiving are associated with current sibling caregiving. Findings also suggest that adults' beliefs about their ill siblings' need for assistance and their parents' need for assistance are related to future sibling caregiving intentions. The implications of these findings for researchers and mental health professionals are discussed. PMID- 11892854 TI - The effects of preparing parents for child psychotherapy on accuracy of expectations and treatment attendance. AB - This study used an experimental design to evaluate the effectiveness of two procedures designed to inform parents about the workings of child therapy, increase the accuracy of their expectations for their child's treatment, and thereby improve their attendance rates. The informational materials explained the importance of parental involvement, how play is used in therapy, confidentiality, and the importance of persisting with treatment until goals are met. Participants were 149 parents or primary caretakers of children aged 3-10 years old. The findings were that: (1) The combination of a brochure and videotape increased the accuracy of parental expectations; (2) the brochure alone had no effect; (3) parents with more accurate expectations had higher rates of treatment utilization on 2 of 7 indices of attendance; and (4) the preparation procedures did not improve attendance rates. Implications for understanding and improving parental utilization of child therapy services are discussed. PMID- 11892855 TI - An investigation of reasonable workplace accommodations for people with psychiatric disabilities: quantitative findings from a multi-site study. AB - Despite the requirement of many employers to provide accommodations in the workplace for individuals with disabilities under Section 504 of the Rehabilitation Act of 1973, the preponderance of accommodations that have been described in the literature concern physical rather than psychiatric disabilities. This study was an exploratory, descriptive, longitudinal, multi site investigation of reasonable workplace accommodations for individuals with psychiatric disabilities involved in supported employment programs. We discuss the functional limitations and reasonable accommodations provided to 191 participants and the characteristics of 204 employers and 22 service provider organizations participating in the study. Implications for service providers and administrators in supported employment programs are discussed. PMID- 11892856 TI - Satisfaction with clinical case management services of patients with long-term psychoses. AB - Outpatients with long-term psychotic disorders from two clinical case management programs were interviewed one-on-one to determine their satisfaction with specific aspects of such services. Patients answered standardized questionnaires and two open-ended satisfaction questions. The close follow-up provided by these programs led us to expect high patient satisfaction, and most patients were very satisfied with their clinical case managers. Lower functioning patients, followed by more intensive clinical case management, were equally satisfied to those followed by a less intensive program. However, many patients revealed dissatisfaction with explanations about clinical treatment and services. PMID- 11892857 TI - Does rehabilitation meet the needs of care and improve the quality of life of patients with schizophrenia or other chronic mental disorders? AB - The effectiveness of a rehabilitation intervention (Boston University Model) was investigated in a one-year prospective naturalistic study among 35 clients with mainly psychotic or affective disorders and dependent on mental health care with at least one hospital admission in the past five years. Rehabilitation was successful in goal-attainment after 1 year (46% fully, 34% partly). Although rehabilitation did not make clients less dependent upon care, it decreased the number of needs and had a positive effect on the match between care needed and care provided. No evidence was found for a significant effect of rehabilitation clients' quality of life and functioning, although social functioning became more in line with the seriousness of psychiatric impairment. PMID- 11892858 TI - The integration of treatment and rehabilitation in psychiatric practice and services: a case study of a community mental health center. AB - After briefly reviewing the relationship of psychosocial rehabilitation to psychiatric practice, the authors recommend a renewed commitment of psychiatrists to bridge and integrate psychiatric treatment with psychosocial rehabilitation in practice and in the organization of services. They use the case example of an urban, community mental health center to illustrate a strategy for achieving greater integration of these two, relatively independent fields of professional practice. The Center's strategy for integration includes (1) center-wide planning, (2) structuring the medical staff office to support the task of integration, (3) establishing a model of practice and principles of care that supports both domains of intervention, (4) educating medical staff about psychosocial rehabilitation, (5) inter-disciplinary team building, including a definition and discussion of professional roles, (6) expanding services research on psychosocial rehabilitation, and (7) advocating in alliance with rehabilitation colleagues for expanded psychosocial rehabilitation services and their integration with treatment. By taking initiative to forward the integration of treatment and rehabilitation, psychiatrists better serve seriously ill patients and more effectively define their own work and roles. PMID- 11892859 TI - Benign breast disease. AB - This article describes 25 years of clinical experience in the setting of a Breast Health Center devoted to benign and malignant disease of the breast. During this period, more than 100,000 patients have been evaluated and treated for a wide variety of breast problems. This experience has provided an extraordinary opportunity to evaluate the natural history of benign breast disease and the frequent observation that the relevant medical literature does not reflect the entire spectrum of these conditions or the appropriate treatment. Most textbooks on breast disease emphasize breast cancer and the late manifestations of benign breast disease that often require surgical treatment. More than 180,000 cases of breast cancer occur each year in the United States. The number of women with benign breast disease is far greater and can be counted in the millions. For these patients, the well-trained primary care physician can provide appropriate evaluation and treatment, including appropriate recommendations for referral. For most patients with breast symptomatology, the goal is relief of symptoms and resolution of the problem. To accomplish this requires a contemporary knowledgebase combined with adequate time spent with the patient. PMID- 11892860 TI - Breast augmentation and breast reduction. AB - Breast enhancement through augmentation improves not only the woman's physical appearance but also contributes to her psychologic well-being. With the current emphasis placed on women's breasts in the media, it is not surprising that small breasted women feel inadequate. Recent US Federal Drug Administration approval of saline implants has given them a new image, and more women are seeking breast augmentation. (27) As long as the woman understands that this operation has associated risks, a physician-patient relationship may be developed resulting in many years of happiness and increased self-esteem. The shape, contour, and size of a woman's breasts are permanently altered by augmentation mammaplasty and breast reduction. Although each case is unique, the outcome of each procedure should result in a satisfied patient. PMID- 11892861 TI - Breast cancer: an introduction to the problem. AB - Breast cancer seemed to be an unsolved problem for much of the last century. An unchanging death rate from 1930 to 1960 occurred in the face of enormous changes in diagnosis and treatment. This article discusses many of the treatments that have been applied, including hormonal manipulation, supervoltage radiotherapy, chemotherapy, and the recently accepted use of tamoxifen as postsurgical treatment. PMID- 11892862 TI - Current status of screening mammography. AB - The results of RCTs conducted around the world indicate that screening mammography can substantially reduce death rates from breast cancer among women aged 40 years and over. Compelling evidence suggests that annual screening should be more effective than screening offered every 1 to 2 years. Annual screening beginning at age 40 years is now recommended by the American Cancer Society, the American Medical Association, and the American College of Radiology* Based on Swedish studies, it is likely that screening mammography can reduce breast cancer deaths by at least 50%. Screening mammography is highly cost-effective and can be performed at acceptable levels of radiation risk and rates of false-positive biopsies. By recommending screening mammography to their patients, the primary care physician can have a pivotal role in reducing the death rate from a major disease of women, similar to the effectiveness of screening for carcinoma of the cervix. PMID- 11892863 TI - Nonpalpable breast lesions: biopsy methods and patient management. AB - Mammography has become a major, if not the best available, diagnostic tool for the early detection of breast cancer. Screening has progressed substantially from the anecdotes of physicians in the early 1970s, that is, the assumption that "if I can't feel it, it's not there." Although controversy continues regarding the earliest age at which screening mammography truly lowers the death rate from breast cancer, the fact that mammography detects breast cancer years before it might be discovered as a mass in the breast cannot be challenged. Mammographic techniques have improved to the point at which smaller and smaller areas of suspicion can be identified, and mammographers have gained greater experience in the interpretation of these minute radiographic abnormalities. The ability to detect these changes has inevitably led to an increase in procedures designed to explain them. The incurred costs, both emotional and economic, of patient recalls for positive mammographic findings are considerable. Regardless of whether the physician practices medicine as a patient advocate or exercises politically correct and cost-effective mandates, the management of nonpalpable breast lesions requires the correlation of cognitive and procedural skills and cooperation among physicians and reflects the technical achievements of contemporary medicine. PMID- 11892864 TI - Risk factors for breast cancer. AB - Breast cancer is multifaceted, and multiple risk factors most likely contribute to each case of the disease. Through further elucidation of highly penetrant autosomal dominant mutations and, perhaps more importantly, weaker polygenic influences, rational therapies to treat or prevent malignancy may develop. Determining the nature and sequence of genetic changes in premalignant breast tissue may offer the greatest opportunity to alter the process of breast cancer development. Perhaps the most difficult challenge is to understand the environmental risk factors that predispose to breast cancer. Although endogenous factors such as hormonal influence on breast cancer risk have been established, this information has not greatly affected our ability to prevent or significantly reduce the risk of disease. National and regional collaborative efforts are needed to fund research directed at defining how the environment and lifestyle factors affect the risk of cancer development. PMID- 11892865 TI - Contemporary management of breast cancer. AB - Breast cancer is a heterogenous disease with significant variations in biologic potential, ranging from small, low-grade, DCIS discovered mammographically with essentially no impact on patient survival to rapidly growing, palpable, locally advanced invasive breast cancer with clinically palpable nodal metastasis. The current challenge is to identify the clinical, pathologic, and molecular factors that determine the biologic potential of a particular breast cancer. Although size, nodal status, histologic grade, age, surgical margin, and hormone receptor status of breast cancer are the most important prognostic factors, the focus of research must be beyond these factors to other nonspecific prognostic information. Bone marrow micrometastasis may be an important factor to help predict outcome (7a) and the complement of sentinel node biopsy, bone marrow analysis, and primary tumor features may allow physicians to better select therapy. With increased understanding of the individual molecular events that control the invasive potential of a particular cancer, practitioners should be better able to predict more accurately which patients have little risk of recurrent disease or metastasis and would be best served by surgery alone versus patients who have a high risk of recurrent and metastatic disease and who should receive multimodality care. PMID- 11892866 TI - The treatment of ductal carcinoma in situ of the breast. AB - As screening mammography has become more frequently used to screen asvmptomatic women, the diagnosis of ductal carcrinoma in situ (DCIS) has become commonplace. Its treatment remains contentious, ranging from mastectomy to local excision alone. The goal of treatment for DCIS is breast conservation, however, as many as 25% of women with this diagnosis may require mastectomy. Although no clear selection criteria have been adopted to subdivide patients into groups best treated by either mastectomy or local excision with or without radiation therapy, many patients with DCIS are candidates for local excision alone, if the biology of the disease is favorable, the size is small, and the margins are negative. Radiation therapy added to local excision decreases the likelihood of recurrence; however, if there is recurrence when first radiation is employed, the patient's only remaining choice often is mastectomy. PMID- 11892867 TI - Adjuvant therapy in breast cancer. AB - The new millennium ushers in an exciting time in the treatment of early stage breast cancer. Although worldwide incidence statistics have not changed significantly in the past decade, mortality rates have declined. This change seems to be related to public health initiatives that increase early detection and awareness and to an increase in the efficacy of adjuvant treatments, including advances in chemotherapy and the emergence of biologic treatments. Physicians from many specialties participate in multidisciplinary tumor boards. Moreover, patients, families, and advocacy groups have taken on new responsibilities, offering encouragement and support for clinical and basic research. PMID- 11892868 TI - Breast reconstruction. AB - This article reviews the most common procedures utilized by plastic surgeons for breast reconstruction. It discusses the different types of autologous reconstruction and modifications of each. When autologous tissue is not an option, reconstruction using an implant is explored. The other breast may require surgery in order to gain symmetry. Surgical options for the opposite breast include breast augmentation, mastopexy, or breast reduction. PMID- 11892869 TI - Nipple discharge. AB - By performing a thoughtful evaluation including a detailed history and careful physical examination, appropriate studies can be selected to allow the practitioner to arrive at the correct diagnosis of nipple discharge in a nonmorbid, expeditious, and inexpensive manner. This article has presented a simple, cost-effective, minimally morbid algorithm for the evaluation of nipple discharge. PMID- 11892870 TI - Breast cancer in pregnancy. AB - Breast cancer in pregnancy is likely to become more common because more women have been waiting to bear children until they are in their 40s. This article presents an overview of pregnancy-associated breast cancer and a review of surgical, chemotherapeutical, and radiation principles as they pertain to pregnancy. PMID- 11892871 TI - Psychosocial issues in breast cancer. AB - The breast cancer experience can bring with it a broad range of emotional sequelae for the woman, her spouse/partner, and her family. Often, patients bear these emotional burdens silently and miss opportunities for the physician and health care team to offer recommendations and interventions for her and her family. The proactive physician evaluating these potential difficulties and using the strength a good relationship with the patient in making appropriate referrals for counseling or support group does much to help the woman embrace as full a recovery as possible. Recommendations presented by the woman's physician are most often heeded. The physician must intervene on behalf of the woman to ensure that psychosocial support needs are met. Physicians are encouraged to adopt this orientation and to use an integrated approach in caring for the woman with breast cancer. (6) PMID- 11892872 TI - The diagnostic evaluation. AB - Diagnostic evaluation is performed to confirm the diagnosis and extent of the disease. The evaluation begins with a careful history and physical examination. Major changes have occurred in the diagnostic evaluation of patients. Frequently the diagnosis is suspected because of an abnormal mammogram or ultrasound, and image-guided biopsies have all but replaced open biopsy. For the primary care physician, the best strategy is to refer the patient to a specialist or a multidisciplinary breast center. PMID- 11892873 TI - Medicolegal considerations in the diagnosis of breast cancer. AB - From the standpoint of the obstetrician, gynecologist, failure to diagnose breast cancer is a significant medicolegal issue in terms of the number of claims initiated and the indemnity awards paid to successful plaintiffs. The incidence of breast cancer may continue to increase. Without quality care and good risk management on the part or health care providers, claims for failure to diagnose cancer in a timely manner will also increase. The challenge for the health care provider is to formulate a plan that promotes early detection and treatment while allowing for independent clinical judgment. Ideally, the plan should be written and followed for every patient in whom complaints of a breast mass have been documented. It is incumbent upon the physician and office staff to create an atmosphere in which patient complaints are not minimized, the limitations of mammography are recognized. follow-up procedures are in place and strictly followed, and a definitive diagnosis within 4 to 6 weeks of the initial presentation is the ultimate goal. Following these suggestions will improve the quality of health care for the patient and significantly decrease the likelihood of litigation alleging a failure to diagnose breast cancer. In the event breast cancer is diagnosed and a lawsuit is brought for failure to diagnose in a timely fashion, the best defense a physician can have is to be able to demonstrate that the patient's complaints were taken seriously, a consistent treatment plan was followed and documented, and the patient received appropriate care. PMID- 11892874 TI - Breastfeeding. AB - A large and growing body of scientific evidence suggests that breastfeeding provides immediate and long-lasting health advantages for the mother and her infant. In the United States, breastfeeding rates currently are the highest recorded in 30 years, although premature weaning owing to the largely avoidable problems of breast pain and concern about adequate milk supply is still common. The advantages of breastfeeding will be more widely appreciated when all health care professionals acquire competence in evidence-based lactation management strategies. These strategies include helping women to position and attach their newborns correctly, encouraging frequent and effective feedings at the breast from birth onward, teaching new parents the signs of adequate milk intake, and providing the resources for promoting breastfeeding without the competition of commercial product promotion. PMID- 11892875 TI - Common breastfeeding problems. AB - Breastfeeding is a learned art. It also is a physiological, psychological, and social event involving two individuals. Many factors impact the breastfeeding experience. This article outlines proper management, good counseling skills, anticipatory guidance, and ongoing support, which can facilitate positive outcomes in breastfeeding. PMID- 11892876 TI - Inflammation of the breast. AB - The primary care physician usually is the first person to see patients complaining of breast pain or nipple discharge. The diagnosis of lactational mastitis is evident because of the history The major consideration is prompt and effective treatment and close follow-up evaluation. Failure to respond to appropriate therapy should suggest abscess formation, and prompt intervention is required. Any diagnosis of mastitis in a patient who is not lactating should be viewed with suspicion. Although several benign and non-life-threatening conditions have been discussed herein, inflammatory breast cancer must always be considered. PMID- 11892879 TI - A fish hook and liver disease: revisiting an old enemy. AB - Vibrio vulnificus is an uncommon but potentially devastating pathogen. Early recognition with prompt antimicrobial therapy and surgical treatment are key factors for a favorable outcome. Patients with diseases of the liver represent the group at highest risk of infection. However, clinicians are often unaware of underlying liver disease in these patients at the time of presentation. We present a case of fulminant V. vulnificus infection in a patient with previously undiagnosed liver disease. PMID- 11892878 TI - Unexpected leukocytosis in a preoperative patient. PMID- 11892880 TI - Widespread ST-segment depression in the electrocardiogram of a 39-year-old woman with chest pain. PMID- 11892881 TI - First 100 cases of gamma knife radiosurgery in Louisiana: analysis of demographics and early results. AB - Louisiana's first ever Leksell Gamma Knife was commissioned at Louisiana State University Health Sciences Center (LSUHSC) in Shreveport in January 2000. Between January 2000 and January 2001,113 patients with various indications were treated using the 201-source Co-60 Leksell model "B" Gamma Knife (Elekta Instruments, Atlanta, Georgia) at LSUHSC-Shreveport. Sixty-three patients were female and 50 were male. The patient age ranged between 13 and 87 years (mean age = 57 years). Fifty-eight (51.3%) patients received radiosurgery as the first line of treatment for their disease, while 55 (48.7%) had previous operations or radiation therapy. The median Karnofsky Performance Score of the patients was 80 (range = 70 to 100). Cerebral metastases were the main indication for radiosurgery at our center accounting for 35% of the patients, while meningioma, arteriovenous malformation, trigeminal neuralgia, and primary central nervous system malignant tumors were the other indications. PMID- 11892882 TI - High dose rate afterloading 192 iridium prostate brachytherapy. AB - Since the evolution of prostate-specific antigen testing for prostate cancer, men have been diagnosed with earlier staged disease. Earlier staged cancer is amenable to local therapy. Over the last decade, new technology has evolved in the treatment of localized prostate cancer. High dose rate afterloading 192 Iridium prostate brachytherapy is a new innovative technique utilizing state of the art equipment and highly specialized computers to achieve precise treatment of prostate cancer with radiation. We present our initial experience with this innovative technique. PMID- 11892884 TI - ECG of the month Here we go, again! ECG of the month. Here we go, again! Atrial tachycardia. PMID- 11892885 TI - Benign paroxysmal positional vertigo. AB - Benign paroxysmal positional vertigo is a disorder of the peripheral vestibular system characterized by brief episodes of vertigo precipitated by head movements in certain planes. It is one of the most common causes of vertigo. Displaced otolithic debris in the posterior semicircular canal is the proposed mechanism of this disorder and explains the success of repositioning procedures in treating these patients. The pathophysiology, diagnosis, and treatment options are discussed. PMID- 11892883 TI - Hydrocephalus. AB - In treating patients with hydrocephalus, cerebral ventricular shunts and their complications are often encountered by the primary care provider. Caring for these patients can provoke anxiety and doubt in those who have had little exposure in dealing with shunts. Becoming familiar with the clinical findings, etiology, and treatment of hydrocephalus, as well as the signs and symptoms of cerebral ventricular shunt complications, will aid the primary care physician in the management, treatment, referral, and continuing care of these patients. PMID- 11892886 TI - Preattentive perception of multiple illusory line-motion: a formal model of parallel independent-detection in visual search. AB - The phenomenon referred to as illusory line-motion (ILM; O. Hikosaka, S. Miyauchi, & S. Shimojo, 1993a) has been described as a measure of the local facilitation of attention gradient. However, J. Kawahara, K. Yokosawa, S. Nishida, and T. Sato (1996) have demonstrated a spatially parallel search for an "odd man out" in the ILM direction. Apart from showing preattentive ILM perception in terms of an analogy between line-motion and apparent motion, the authors examined whether ILM perception is possible without attention from another point of view. Four experiments revealed that the ILM target can be detected in parallel without invoking attentional facilitation and invalidated the possible contribution of attentional set in parallel ILM search. Participants were able to correctly detect the ILM target among multiple nontargets, even when the line orientation was changed from trial to trial. The authors' independent detection model predicted ILM search performance well on several occasions. These findings strongly support a preattentive and stimulus-driven explanation of ILM perception. PMID- 11892887 TI - Monaural loudness adaptation for middle-intensity middle-frequency signals: the importance of measurement technique. AB - Using the Simple Adaptation technique (SA) and the Ipsilateral Comparison Paradigm (ICP), the authors studied monaural loudness adaptation to a middle intensity [60 dB(A)] tone at signal frequencies of 250, 1000, and 4000 Hz in the left and right ears. Adaptation effects were absent when the SA procedure was used. However, they were observed uniformly across all frequency values with the ICP, a result that challenges the assertion in the literature, on the basis of SA measures, that loudness adaptation for middle-intensity signals occurs only at frequencies above 4000 Hz. The ICP features periodic intensity modulations (+/-10 dB relative to the base signal) to accommodate listeners' needs for referents by which they can gauge subtle changes in the loudness of the adapting tone, a key component that is missing in the SA method. Adaptation effects in this investigation were similar in both ears, supporting the equal susceptibility assumption common in loudness adaptation studies. PMID- 11892888 TI - Melatonin and sleep qualities in healthy adults: pharmacological and expectancy effects. AB - The impact of expectancy on melatonin's effects on sleep qualities was investigated. Both the pharmacological dose of 6 mg of melatonin and the expectation of receiving melatonin were predicted to improve subjective ratings of sleep qualities. The balanced placebo design varied 2 factors within-subjects: actual treatment and expected treatment. Adults (N = 53; 21 men and 32 women) between the ages of 26 and 71 years were administered either 6 mg of melatonin or a placebo for 8 nights. An instructional manipulation directed participants' expectations. Participants rated their nightly sleep experiences. Results revealed that feelings upon awakening differed between genders and that expecting melatonin increased ratings of sleep continuity. Most important, high ratings of "grogginess/tiredness" were associated with receiving melatonin, regardless of expectancy, as well as with receiving placebo when melatonin was expected. Overall, the findings underscore the need to consider expectancy and gender differences in research on melatonin and sleep experiences. PMID- 11892889 TI - Anxiety and repression in attention and retention. AB - High- and low-anxious college students (as determined by scores on the Taylor Manifest Anxiety Scale; A. W. Bendig, 1956) and repressors (low anxiety and high scores on the Marlowe-Crowne Social Desirability Scale; D. P. Crowne & D. Marlowe, 1964) were compared on 3 cognitive tasks. High-anxious participants more often spelled the negative emotional meaning of ambiguous homophones (e.g., pane/pain) and forgot more of their free associations to emotional cue words than did the low-anxious participants. The repressors also detected the emotional meaning of the homophones but unlike the anxious, the repressors did recall their associations to the emotional words. In a working memory task using nonemotional items, the moderately anxious participants recalled fewer words than did the low- and high-anxious participants. The results confirm that both trait anxiety and repression affect information processing at a variety of stages but not in the same way. Repressors were sensitive to, and retentive of, negative emotional stimuli. PMID- 11892890 TI - The influence of learning style and cognitive ability on recall of names and faces in an older population. AB - The present study examined whether, for older adults, a verbal or imagery cognitive style is associated with recall of names and faces learned in an experimental condition. Cognitive abilities that are represented in current models of face recognition and name recall were also examined. Those abilities included picture naming, verbal fluency (i.e., naming items within a given category), vocabulary comprehension, visual memory, and the learning of unassociated word pairs. Fifty older adults attempted to learn first and last names of 20 student actors and actresses pictured on videotapes (40 names total). On average, participants learned the most first names, followed by last names, and the fewest full names. The greater the number of responses on a questionnaire associated with an imagery cognitive style, the more the names of faces were correctly identified by participants. There was no significant relationship between a verbal cognitive style and the number of names and faces recalled. As for cognitive abilities, all of the abilities measured--with the exception of vocabulary comprehension--were significantly associated with the number of names and faces learned. A regression analysis indicated that the best predictor of successful name-face learning was the participants' ability to learn and recall 5 unrelated word pairs. When that cognitive measure was deleted from the regression analysis, delayed visual memory and verbal fluency were the next best predictors of the older adults' ability to learn names and faces. PMID- 11892891 TI - The nature of declarative and nondeclarative knowledge for implicit and explicit learning. AB - Using traditional implicit and explicit artificial-grammar learning tasks, the author investigated the similarities and differences between the acquisition of declarative knowledge under implicit and explicit learning conditions and the functions of the declarative knowledge during testing. Results suggested that declarative knowledge was not predictive of or required for implicit learning but was related to consistency in implicit learning performance. In contrast, declarative knowledge was predictive of and required for explicit learning and was related to consistency in performance. For explicit learning, the declarative knowledge functioned as a guide for other behavior. In contrast, for implicit learning, the declarative knowledge did not serve as a guide for behavior but was instead a post hoc description of the most commonly seen stimuli. PMID- 11892892 TI - Adenocarcinoma of the colon presenting as polymyositis: case report. AB - The relationship between polymyositis and malignancy is well known. Several types of tumors can be complicated with myositis and/or more frequently dermatomyositis. It has been suggested that tumors of the large bowel are rarely complicated by myositis. We describe a patient with adenocarcinoma of the colon presenting as polymyositis and review the available literature. PMID- 11892893 TI - Comparative antimicrobial spectrum and activity of BMS284756 (T-3811; a desfluoroquinolone) tested against an international collection of staphylococci and enterococci, including in vitro test development and intermethod comparisons. AB - The study was initiated to determine the in vitro activity and MIC/disk test comparisons of BMS284756, a new des-fluoro(6)-quinolone, against isolates of staphylococci and enterococci from the SENTRY Antimicrobial Surveillance Program, 2000. Isolates were tested by reference broth microdilution and standardized disk diffusion methods. Against 3,789 strains of gram-positive cocci from the SENTRY Program (2000), the BMS284756 MIC90 and percentage susceptible at < or = 2 and < or = 4 microg/ml were: Staphylococcus aureus (4 microg/ml; 89.3 and 97.1%), coagulase-negative staphylococci (CoNS; 4 microg/ml; 86.1 and 96.0%) and enterococci (> 4 microg/ml; 62.0 and 76.2%). Also tested were selected staphylococci (300 strains) and enterococci (102 strains) by two standardized methods. The activity of BMS284756 was highly correlated with oxacillin resistance among staphylococci. Oxacillin-susceptible staphylococci were all inhibited by BMS284756 at < or = 0.5 microg/ml, whereas oxacillin-resistant strains required inhibitory concentrations of > or = 1 microg/ml. Excellent correlation was observed between the MIC and 5-microg disk zone diameter for staphylococci and enterococci (r=0.91 to 0.93). Among vancomycin-susceptible enterococci, 67% of Enterococcus faecalis, 25% of E. faecium, and 76% of other Enterococcus spp. isolates were inhibited by BMS284756 at < or = 2 microg/ml. All vancomycin-resistant enterococci (VRE; 11 E. faecalis and 15 E. faecium) were inhibited by > or = 2 microg/ml of BMS284756. Among the non-VRE, non-faecium enterococcal isolates (n=64), 62% were inhibited by < or = 0.5 microg/ml. BMS284756 showed excellent activity against oxacillin-susceptible staphylococci and moderate activity against enterococci other than VRE and E. faecium. Acceptable correlations were observed between MIC and disk test results for both tested genus groups. PMID- 11892894 TI - Moxifloxacin sensitivity of respiratory pathogens in the United Kingdom. AB - The in vitro activity of moxifloxacin and comparator agents against respiratory isolates from a range of geographically distinct centres around the United Kingdom was investigated in the following study. Clinical isolates of Streptococcus pneumoniae (n = 257), Haemophilus influenzae (n = 399) and Moraxella catarrhalis (n = 253) were obtained between March 1998 and April 1999 from nine centres in the United Kingdom. Sensitivity was determined by testing each isolate for its minimum inhibitory concentration (MIC) by agar dilution. Against Streptococcus pneumoniae moxifloxacin and grepafloxacin were the most active (MIC90 = 0.25 mg/l). Trovafloxacin and sparfloxacin were the next most active (MIC90 = 0.5 mg/l) followed by levofloxacin and ciprofloxacin. MIC90 values of the six fluoroquinolones versus H. influenzae ranged from <0.0039 mg/l to 0.0625 mg/l and from <0.0039 mg/l to 0.5 mg/l for M. catarrhalis. The rank order of activity of the fluoroquinolones versus H. influenzae was moxifloxacin = trovafloxacin = grepafloxacin = sparfloxacin > ciprofloxacin > levofloxacin. Against M. catarrhalis the lowest MIC90 was that of grepafloxacin at 0.0625 mg/l followed by moxifloxacin, sparfloxacin, levofloxacin and ciprofloxacin. Trovafloxacin demonstrated the highest MIC90 at 0.5 mg/l. These results demonstrate that moxifloxacin has superior in vitro activity against respiratory tract pathogens than any other comparator quinolones available for clinical use. PMID- 11892895 TI - Antimicrobial resistance in gram-negative isolates from European intensive care units: data from the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) programme. AB - Susceptibility data were collected for 6243 gram-negative isolates from 29 European ICUs participating in the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Programme (1997-2000). The most commonly isolated bacteria were Pseudomonas aeruginosa (22.5%), Escherichia coli (19.8%), Klebsiella pneumoniae (10.4%), and Enterobacter cloacae (7.7%). The incidence of extended-spectrum beta-lactamase-producers was higher in Turkish, Russian and Italian ICUs (27.9-39.6%) than in other countries (2.5-10.8%). The frequency of AmpC-cephalosporin hyper-producers was 16.8-55.4%. Meropenem was more active against Proteus mirabilis than imipenem (99.0% versus 88.8% susceptibility, respectively). Against Acinetobacter baumannii, meropenem (79.6% susceptible) and imipenem (82.2%) were more active than comparators (34.3-51.6%). Meropenem and imipenem exhibited good activity against P. aeruginosa (76.1% and 68.2%, respectively; but with inter-country variation). Ciprofloxacin resistance in E. coli and K. pneumoniae increased and needs close monitoring. Meropenem (98.2 99.8% susceptibility) and imipenem (88.8-99.4%) remained potent against important species of gram-negative bacteria from European ICUs actively using meropenem. PMID- 11892896 TI - Immunomodulating activity of quinolones: review. AB - Fluorinated quinolones exert their bactericidal activity by inhibiting bacterial type II topoisomerases. At therapeutic concentrations, quinolones superinduce interleukin-2 (IL-2) and interferon-gamma production by mitogen-activated human peripheral blood T lymphocytes. At the molecular level, a stronger activation of the nuclear factor AP-1 ('activator protein-1') is observed in cells incubated with ciprofloxacin, resulting in enhanced cytokine gene transcription. Several cytokine and immediate early (e.g., c-fos and c-jun) mRNAs are upregulated by ciprofloxacin, possibly reflecting a mammalian stress response. In cultures with murine splenocytes, quinolones enhance IL-3 and granulocyte-macrophage colony stimulating factor (GM-CSF) synthesis. The stimulation of these hematopoietic growth factors prolongs survival of mice with depressed bone marrow and prevents experimental antiphospholipid syndrome (APS). In contrast, quinolones inhibit both human and mouse monocytic IL-1 and TNF-alpha synthesis, an effect that is beneficial in rat experimental type II collagen induced arthritis and LPS-induced septic chock in mice. The intriguing immunomodulatory activities of fluoroquinolones warrant future investigations with new tailored derivatives. PMID- 11892897 TI - A 10-year survey of Mycobacterium tuberculosis isolates in Pavia and their drug resistance: a comparison with other Italian reports. AB - A retrospective review was made of the bacteriological and medical records of patients with culture-confirmed pulmonary tuberculosis who attended the IRCCS San Matteo Polyclinic of Pavia, between 1990 and 2000. Altogether, 279 patients were included in the survey: 220 new cases and 59 prior treatment cases. Resistance to at least one drug, and resistance to both isoniazid and rifampicin (MDR) were more common among previously treated patients than among new cases (86.4% vs. 34.1%, and 44% vs. 5.9%, respectively). While the frequency of resistance to any drug showed no variation in the period examined, a trend toward a progressive decrease in the frequency of primary MDR-TB was observed (from 11.9% in 1990-1992 to 1.3% in 1998-2000). The level of resistance observed in our study suggests that all isolates of Mycobacterium tuberculosis should be tested for drug susceptibility, especially when obtained from patients who report a previous episode of the disease. PMID- 11892898 TI - Staphylococcus aureus and Escherichia coli adhesion to human cells is reduced by sub-MICs of gemifloxacin. AB - This study was designed to investigate the capacity of subinhibitory concentrations of the newly developed fluoroquinolone antibiotic gemifloxacin to interfere with the mechanism of bacterial adhesion. Human buccal epithelial cells were incubated with Staphylococcus aureus and Escherichia coli, and grown in the presence of serial dilutions of gemifloxacin from 1/2 MIC to 1/128 MIC. A significant decrease in the adhesion of both S. aureus and E. coli was observed from 1/2 MIC to 1/32 MIC. Morphological changes including filamentous forms of E. coli and cluster formation and swelling of S. aureus were also observed, mainly from 1/2 MIC to 1/8 and 1/16 MIC. These findings are discussed in terms of dose effect relationships and the interpolation of this pharmacodynamic data with the pharmacokinetics curve of gemifloxacin. PMID- 11892899 TI - Experimental studies on synergism between aminoglycosides and the antimicrobial antiinflammatory agent diclofenac sodium. AB - The antiinflammatory agent diclofenac sodium (Dc) exhibited remarkable antibacterial effects both in vitro and in vivo. Fifteen different bacteria sensitive to Dc as well as to a number of common antibiotics were tested for synergistic effects in vitro. Disc diffusion test with Dc and aminoglycosides assessed by stringent computation showed clear-cut synergism. Synergism between Dc and streptomycin (Sm) was found to be statistically significant (p < or = 0.01) when compared with their individual effects. By the checkerboard assessment procedure, the fractional inhibitory concentration (FIC) index of this combination was found to be 0.49, confirming synergism. The mouse protective capacity of this combination was then evaluated in vivo against S. typhimurium as the virulent infecting bacterium, and the size of bacterial load determined from infected autopsied animals. Statistical analysis by Student's 't' test suggested this drug combination is highly synergistic; synergism was also noted between Dc and other aminoglycosides. PMID- 11892900 TI - Penetration of piperacillin/tazobactam (4 g/500 mg) into synovial tissue. AB - The degree of penetration of an antibiotic into the infected site is an important criterion for therapeutic success. This is particularly true for bone and joint infections. The association of piperacillin and tazobactam has been widely used in the treatment of serious infections including bone infections, but no study has been devoted to the subject of its diffusion into synovial tissue. Our objective was to quantify piperacillin/tazobactam synovial tissue penetration and to estimate the efficacy of the association against the microorganisms usually encountered in joint infections. In an open-label study, 6 subjects with similar age, weight, height and creatinine clearance, who were undergoing elective total hip replacement, received a single, parenteral, 4 g/500 mg dose of piperacillin/tazobactam. Plasma and synovial tissue samples were collected and analyzed by a validated HPLC method. The mean concentrations of piperacillin and tazobactam 1.5 h after the initiation of infusion were 69.9 +/- 4.9 microg/mL and 7.7 +/- 0.3 microg/mL, respectively, in plasma and 37.1 +/- 2.1 microg/g and 2.8 +/- 0.4 microg/g, respectively, in synovial tissue. The synovial tissue/plasma ratios were 0.5 +/- 0.0 for piperacillin and 0.4 +/- 0.0 for tazobactam. The piperacillin/tazobactam ratios were 9.1:1 in plasma and 13.5:1 in synovial tissue. The concentrations achieved in synovial tissue are above the MICs of most of the susceptible pathogens usually involved in joint infections, which suggests that the piperacillin/tazobactam combination should be effective in the treatment of most joint infections caused by susceptible microorganisms. PMID- 11892901 TI - Questionnaire survey of perioperative antibiotic prophylaxis in Italian surgical departments. AB - Correct antibiotic prophylaxis reduces the incidence of postoperative infections. 600 questionnaires on perioperative antibiotic prophylaxis were sent to Italian Surgical Departments. Each questionnaire included a series of 17 multi-choice questions concerning the specific approach of the department to: organization, type, timing, duration, auditing of prophylaxis. 435 departments (72.5%) responded to the questionnaire; 50 of these were blank, so 385 out of 435 (88.5%) were suitable for statistical evaluation. Results were as follows: 90.5% of departments perform some form of prophylaxis under the control, in 90.5% of cases, of surgeons; 89.3% differentiate antibiotics according to class of operation; 67.4% give the antibiotic preoperatively and prefer i.v. injection (61.0%), mostly in the ward (56.2%); in 33.3% of cases the prophylaxis is standard (more than 2 doses), but 55.8% of Italian surgeons do not give a boost dose in operations longer than 3 h; 54.2% of patients receive a cephalosporin (mostly III generation), with a rotation of molecules in 53.9% of cases; 71.7% of departments register the incidence of infections, but only 43.2% control the patients 30 days after surgery; finally, 54.2% of departments work together with a bacteriology laboratory active 24 hours, while in 81.7% of cases the hospital has an Infection Committee which meets together usually without a programmed date (60.3%). In conclusion, antibiotic prophylaxis in Italian Surgery Departments appears adequate, even though some problems still remain regarding time-dose duration-schedule, rotation of molecules, excess of cephalosporins, availability of a 24-h bacteriological laboratory and infection surveillance after discharge. PMID- 11892902 TI - Antibiotic prescribing for dental conditions: a community-based study in southern Italy. AB - The aim of this study was to investigate for which conditions antibiotics are being used in community dental practice, and which clinical features represent the most common reason for an antibacterial approach to the treatment of dental conditions. The study was carried out from November 1998 to June 1999. Dentists were selected according to the different areas of southern Italy, from a list provided by the Italian Society of Dentists. Out of 87 selected dentists, 33 agreed to participate and filled in 1615 questionnaires for each therapeutic intervention ending with antibiotic treatment. Analysis of data indicated that alveolar-gingival abscesses were the most commonly treated infection, accounting for 23.6% of total treatments, followed by acute periodontitis (20.6%) and disodontiasis of the 3rd molar (18.5%). Parenteral antibiotics were chosen in 7.8% of cases. Penicillins were the most commonly used group, 40.1% of total treatments, followed by macrolides (30.2%) and cephalosporins (13.4%). Moreover, penicillins were widely used for post-surgery therapy (52.1%) and disodontiasis of the 3rd molar (50.8%), while macrolides were the most commonly used group for gingivitis (44.1%) and parodontal diseases (55.0%). The choice of parenteral antibiotics was related to severe general symptoms (odds ratios [OR], 4.4; 95% CI: 2.2-9.0), pain (OR, 2.7; 95% CI: 1.2-6.1) and lymphonodal involvement (OR, 6.4; 95% CI: 2.7-15.1). In conclusion, our study demonstrates that antibiotic treatment is often based on the eradication of as many microorganisms as possible, and on the clinical assessment of the patients, rather than on any knowledge of the pathogens involved. PMID- 11892904 TI - Netilmicin effect on urinary retinol binding protein (RBP) and N-acetyl-beta-D glucosaminidase (NAG) in preterm newborns with and without anoxia. AB - The study aim was to evaluate urinary excretion of Retinol Binding Protein (RBP), compared with urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG), in preterm infants with anoxia and netilmicin treatment. Urinary RBP and NAG were evaluated in 83 preterm newborns divided in 4 groups: 37 healthy preterm newborns (controls); 14 with neonatal anoxia; 16 treated with ampicillin + netilmicin; 16 with neonatal anoxia and treated with ampicillin + netilmicin. RBP was determined by an automated nephelometric technique and NAG by a colorimetric method on 5-h urine samples in the first week of life. Results showed that urinary excretion of RBP (average from first week values) was 1.06+/-0.67 g/mol creatinine (mean +/- SD) in controls, 1.99+/-1.41 in antibiotic-treated newborns, 3.99+/-4.57 in anoxic newborns and 3.75+/-3.48 in anoxic newborns under antibiotic treatment. When gestational age was not considered, a marked effect of anoxia (P<0.001) and a borderline effect of netilmicin (P<0.059) on RBP excretion were detected by ANOVA. However when gestational age was also considered by analysis of covariance, it appeared as the strongest predictor of RBP excretion (P<0.001), while the effect of netilmicin was no longer significant (P=0.181). The effect of anoxia persisted, although less remarkable (P=0.010). Conversely anoxia did not affect urinary NAG excretion, which was rather correlated with gestational age and netilmicin administration. The authors conclude that RBP and NAG urinary excretion may be used to discriminate between neonatal anoxia and netilmicin treatment, respectively as etiologic factors of renal tubular damage in the newborn. PMID- 11892903 TI - Three-times weekly teicoplanin in the outpatient treatment of acute methicillin resistant staphylococcal osteomyelitis: a pilot study. AB - Treatment of osteomyelitis requires prolonged hospital stay, lengthy antibiotic therapy and adequate surgical debridement. Outpatient parenteral antibiotic therapy (OPAT) is a new approach to reduce patient discomfort and hospital costs. Teicoplanin, a glycopeptide antibiotic with a long half-life (72 hours), is one of the most useful drugs for OPAT. We performed a pilot study to assess the safety and efficacy of three-times weekly teicoplanin in the treatment of methicillin-resistant (MR) acute staphylococcal osteomyelitis. Ten patients with acute post-traumatic osteomyelitis were enrolled. Pathogens were MR Staphylococcus aureus (5 patients) and MR coagulase-negative staphylococci (5 patients). After a loading dose of 400 mg b.i.d. for 3 days, patients were treated with an intravenous dose of 1000 mg on Mondays and Wednesdays and with a 1200 mg dose on Fridays. Teicoplanin trough levels were maintained within a 10 to 20 mg/L range. If hardware removal had been possible at enrollment, treatment was carried out for at least 4 weeks. If, on the contrary, hardware removal had not been possible, teicoplanin was administered as suppressive therapy until hardware removal. Treatment was successfully performed in 9 out of 10 patients, whereas in one patient only improvement was achieved. Side effects were not recorded. Three times weekly teicoplanin seems to be a valuable option in the treatment of acute MR staphylococcal osteomyelitis. Further studies are warranted in order to better define the role of this new administration schedule in this field. PMID- 11892905 TI - Retrospective comparison of single-agent chemotherapy with weekly 5-fluorouracil or weekly irinotecan in previously treated patients with metastatic colorectal cancer. AB - This study is a retrospective analysis of response, toxicity and freedom from progression of two single-agent chemotherapy regimens in patients with previously treated metastatic colorectal cancer. Thirty-five patients with histologically confirmed measurable metastatic colorectal cancer received chemotherapy after failure of first-line 5-fluorouracil (5-FU) and leucovorin treatment. The median age was 61 years. Twenty-seven patients had liver metastases, 6 had local recurrence, 1 had retroperitoneal lymph node metastases and 1 had lung metastases. Eighteen patients received weekly 2600 mg/m2 5-FU and 17 patients received weekly 125 mg/m2 irinotecan (CPT-11). Treatment was given until disease progression. Total number of cycles was 202 for 5-FU and 248 for CPT-11. The relative dose intensity was 1.0 for 5-FU and 0.84 for CPT-11. No grade 3-4 toxicity was registered in patients who received 5-FU. Grade 3-4 toxicity rates were as follows in those who received CPT-11: vomiting 1 (5.9%) patient in 1 cycle, diarrhea 3 (17.7%) patients in 3 cycles and neutropenia in 3 (17.7%) patients in 3 cycles. No patients manifested febrile neutropenia. Two patients (11.8%) needed hospital admission because of toxicity: 1 for vomiting and 1 for diarrhea. No objective responses were observed in the 5-FU group of patients. Three patients (17.6%) who received CPT-11, achieved partial response with a median duration of 8 months. Stable disease was registered in 3 (17.6%) and 9 (52.9%) patients in 5-FU and CPT-11 groups respectively (p=0.05). Median time to progression was 3.3 months for patients who received 5-FU and 4.2 months for those treated with CPT-11 (not significant). One-year survival was 22.2% and 54.3% respectively (p=0.05). CONCLUSION: Weekly chemotherapy with CPT-11 is tolerated with acceptable toxicity and leads to a better response rate than weekly high dose 5-FU. It also significantly improves survival but does not prolong freedom from progression. PMID- 11892906 TI - Brucellosis in the etiology of febrile neutropenia: case report. AB - Brucellosis is one of the leading diseases in the differential diagnosis of fever of unknown origin in some parts of the world. It can lead to treatment failure because of slow growth in blood cultures and late appearance of signs and symptoms in patients with febrile neutropenia who were unresponsive to empirical antibiotic treatment. During the last year in our oncology unit adjuvant chemotherapy was given to 3 patients with breast (n=1) and stomach cancer (n=2) and febrile neutropenia was seen after the first course of chemotherapy (cyclophosphamide, methotrexate, 5-fluorouracil, etoposide, Adriamycin, and cisplatin) in all 3 patients. Cefepime and amikacin were commenced but the fever continued. Prior to antifungal treatment, the patients were re-evaluated because of the history of unpasteurized milk ingestion without overt signs and symptoms. Serum agglutination tests of brucellosis were performed and were 1:640 in two patients and 1:320 in the third. Brucella melitensis was identified only in one case although multiple blood cultures were taken from all 3 patients. Empiric antibiotic treatment was stopped and streptomycin 1 g/day (10 days), doxycycline 200 mg/day (28 days), trimethoprim 320 mg and sulfamethoxazole 1600 mg/day (28 days) were given. Although neutropenia continued, fever subsided in 3 days. Due to high incidence of brucellosis in some geographic areas, especially in the Middle East, brucellosis should be kept in mind in the differential diagnosis of febrile neutropenia. PMID- 11892907 TI - Respiratory distress due to endotracheal aspergilloma in an immunocompromised lymphoma patient: case report. AB - A 54-year old woman, intensively treated for aggressive, relapsed lymphoma had symptoms of severe dyspnea and hoarseness. The diagnosis of endotracheal aspergilloma was made by sputum culture, bronchoscopy and biopsy. The lesions consisted of endotracheal aspergilloma associated with tracheal obstruction due to the mass effect. The patient improved dramatically after removal of the mass. PMID- 11892908 TI - Long-term complete remission of oral cancer after anti-neoplastic chemotherapy as single treatment modality: role of local chemotherapy. AB - The impact of intra-arterial local chemotherapy on squamous cell carcinomas of the oral cavity is doubtful when considering long-term survival, especially in cases of nodal involvement. But even in patients with strictly local disease it is not possible to determine the effect of intra-arterial chemotherapy because it is mainly used as a neoadjuvant treatment modality. In the present paper, long term courses of two patients are described who refused any further treatment after one cycle of intra-arterial chemotherapy with cisplatin followed by systemic chemotherapy with 5-fluorouracil and one cycle of intra-arterial chemotherapy with high-dose cisplatin, respectively. The aim of the paper is to demonstrate the potential of local chemotherapy in responders. The impact of this treatment modality in incurable patients is discussed, too. This may offer a point in favor of use of intra-arterial chemotherapy in combination treatment regimens. PMID- 11892909 TI - Delivering novel targets and antibiotics from genomics. AB - Recent advances in DNA sequencing technology have made it possible to elucidate the sequences of the entire genomes of pathogenic bacteria and concomitant advancements in bioinformatic tools have driven comparative studies of these genome sequences. These evaluations are significantly increasing our ability to make valid considerations of the limitations and advantages of particular targets based on their predicted spectrum and selectivity. In addition, developments in gene-essentiality technologies amenable to pathogenic organisms liave enabled new genes and gene products critical to bacterial growth and pathogenicity to be uncovered at an unprecedented rate. This review will describe how aspects of the above capabilities are impacting the discovery and characterization of known and novel antibacterial targets using specific examples taken from a variety of important, diverse bacterial processes. PMID- 11892910 TI - THF carbapenems. Wyeth Ayerst. AB - CL-191121 is one of three new tetrahydrofuranyl (THF) carbapenems, which possess stability to renal dihydropeptidase and beta-lactamases [162484], [387167], [418244] and have minimal binding to penicillin-binding proteins (PBPs) [163188]. Of the series of THF carbapenems, CL-191121 had the best activity against gram positive organisms, particularly against Enterococcus faecalis. It demonstrated moderate oral activity against an Escherichia coli infection in mice and was 20 fold more efficacious than imipenem [253186]. As the effective oral dose (ED50) was 11- to 14-fold higher than the effective subcutaneous dose, a series of bis double ester prodrugs (of the aminomethyl-THF 1beta-methyl-carbapenems) was prepared, which showed substantially improved oral bioavailability in the mouse model (EU50 = 0.42 microg/ml following a single oral dose obtained for the most potent compound) [258726]. OCA-983 is a prodrug derivative of CL-191121 sharing potent inhibitory activity against class A and class C beta-lactamases [324244], although development of this compound is no longer being pursued. PMID- 11892911 TI - Oritavancin. Eli Lilly & Co. AB - Oritavancin (LY-333328), a lead glycopeptide from a series targeted at vancomycin resistant bacteria, especially enterococci, is under development by Eli Lilly for the potential treatment of bacterial infections. It entered phase III trials in the US in January 2001 [396223] and the company expects to submit an NDA by 2003 [396478]. Oritavancin has been reported to have activity comparable to that of vancomycin and teicoplanin (Aventis Pharma AG), but retains activity against glycopeptide-resistant bacterial strains [407519]. The bactericidal activity of oritavancin suggests that it may prove useful as a single agent therapy in the treatment of antibiotic-resistant enterococci. Although its mechanism of action is unclear, dimerization of the glycosyl portion stabilizing D-Ala-D-Ala binding has been suggested [337644]. PMID- 11892912 TI - Novel therapies in the treatment of systemic lupus erythematosus. AB - This review highlights the advances made in the use of biological and immunomodulatory agents in systemic lupus erythematosus. Although there have been disappointments (eg, anti-CD40 ligand, DNAse), it is clear that DHEA and mycophenolate mofetil will have a place in the management of the disease. In our opinion, leflunomide and LJP-394 are the most promising therapies currently, under study. PMID- 11892913 TI - Angiogenesis: a therapeutic target in arthritis. AB - A variety of pharmacological strategies are being subjected to clinical trial to inhibit neovascularization of solid tumors. Increased angiogenesis is also a key component of synovitis and bone modeling in arthritis. Molecular mechanisms and pathological consequences of blood vessel growth in arthritis are now being elucidated. Preclinical studies of angiogenesis inhibitors in animal models of inflammatory arthritis support the hypothesis that inhibition of neovascularization may reduce inflammation and joint damage. Clinical data are consistent with these models being predictive of efficacy in rheumatoid arthritis. However, controlled studies of specific anti-angiogenic agents in human arthritis remain limited. Further studies are required to demonstrate that pharmacological agents can effectively inhibit articular angiogenesis, and ameliorate inflammation and subsequent joint damage. Potential toxicity of angiogenesis inhibitors in reproduction, growth and development and wound repair may be circumvented by short-term or local application, or by targeting molecular mechanisms that are specific to pathological rather than physiological angiogenesis. PMID- 11892914 TI - Arthritic diseases: melanocortin type 3 receptor agonists as potential therapeutics. AB - Gouty arthritis is currently treated with drugs that have an array of side effects. Therefore, identification of novel endogenous targets for drug development may have beneficial properties ACTH4-10, a heptapeptide fragment derived from the hormone adrenocorticotrophin (ACTH) modulates the inflammatory response in a corticosterone-independent manner, via agonism at melanocortin type 3 receptors (MC3-R) expressed on peritoneal macrophages. MC3-R agonists inhibit cytokine formation and subsequent neutrophil migration, while antagonists abrogate these effects. Together, these data highlight MC3-R as a potential therapeutic target and suggest that small molecule agonists directed at MC3-R with more specific actions, may be potentially novel therapeutics for treating this pathology. PMID- 11892915 TI - VX-745. Vertex Pharmaceuticals. AB - VX-745, a lead anti-inflammatory candidate, small-molecule inhibitor of mitogen activated protein kinase (MAPK), is under development by Vertex Pharmaceuticals Inc in association with Kissei Pharmaceutical Co Ltd for the potential treatment of rheumatoid arthritis (RA) [214928]. VX-745 was introduced by Vertex as a potential antiinflammatory drug for the treatment of RA in a pilot phase II trial initiated in November 1999 [346067]. In June 2000, phase II trials were still ongoing [371819] and in January 2001, Vertex initiated a randomized, double blind, placebo-controlled phase II trial in adult patients with RA, with the objective of evaluating clinical response rates, self-reported patient health assessments and pharmacodynamic markers of drug activity [395083]. During the 33rd Annual Meeting of the American Chemical Society in May 2000, VX-745 was reported to be active against several isotypes of p38 MAPK, including p38alpha, p38beta and p38gamma [368149]. The targeting of p38 MAPK by VX-745 was associated with the suppression of the release of inflammatory mediators, including interleukin (IL)-1beta and tumor necrosis factor (TNF)alpha, known to be implicated in exacerbating the pathophysiology of RA [273648], [368149], [371548], [372054], [408713]. PMID- 11892916 TI - Overview: from heparin to low molecular weight heparin: beyond anticoagulation. PMID- 11892917 TI - The potential of erythropoietin and related strategies to stimulate erythropoiesis. AB - During the 15 years since the cloning of the erythropoietin (EPO) gene, the recombinant hormone has become an extraordinarily successful therapeutic for the treatment of renal and several nonrenal anemias. Considerable insight was gained into the adverse consequences of chronic reduction in hemoglobin levels on quality of life, physiological functions and survival. Precise cost-benefit relationships, however, are still lacking and target hemoglobin levels remain controversial. Nevertheless, given the wide application and potential of erythropoietic stimulation, there is growing interest in alternative but related strategies, including the development of second generation molecule EPO-mimetics, modulators of receptor activity and EPO gene therapy. PMID- 11892918 TI - Antiplatelet therapies: platelet GPIIb/IIIa antagonists and beyond. AB - Accumulating evidence supports the critical role of platelet involvement in arterial thrombosis, and argues for the development of more efficacious, yet safe, antiplatelet therapies. Aspirin continues to be used routinely for the management of acute myocardial infarction, unstable angina and secondary prevention of ischemic events. Nevertheless, adverse clinical outcomes still occur. Newer-generation drugs such as clopidogrel (an adenosine-diphosphate receptor antagonist) and intravenous glycoprotein (GP)IIb/IIIa antagonists (inhibitors of platelet aggregation irrespective of the stimulus) have demonstrated significant clinical benefit. This review will discuss the role of the aforementioned antiplatelet therapies in thrombotic disorders as well as future directions in the field. PMID- 11892919 TI - YM-337. Yamanouchi. AB - Yamanouchi in collaboration with Protein Design Labs (PDL), is developing YM-337, a humanized antibody (C4G1) for the potential treatment of coronary artery disease, including myocardial infarction, arterial thrombosis and ischemic stroke. The antibody is based on a Yamanouchi murine antibody specific for the glycoprotein (GP) IIb/IIIa receptor found on human platelets 11942421. By June 1999, Yamanouchi was carrying out phase II trials on YM-337 in high-risk percutaneous transluminal coronary angioplasty (PTCA) patients in Europe and the US [326891]. These were ongoing in May 2001 [409092], [411763]. Phase II trials for the potential treatment of ischemic stroke were also underway by this time in Europe and the US [411763]. By 1998, Yamanouchi and PDL had an agreement for the development of a SMART C4G1. Yamanouchi holds exclusize worldwide rights to YM 337 and is responsible for all clinical trials and obtaining regulatory approval. As part of the collaboration, PDL has received milestone payments for humanization of the antibody and is entitled to receive royalties on product sales [274540]. PMID- 11892920 TI - Darbepoetin alfa. Amgen. AB - Amgen has launched darbepoetin alfa, synthetic recombinant novel erythropoiesis stimulating protein (NESP), for the treatment of anemia associated with renal disease. The drug was approved by the European Commission in June 2001, under the tradename Aranesp, for the treatment of anemia in chronic kidney failure including patients on and not yet on dialysis. The company launched the drug on day one of its approval in the following countries: Germany, Sweden, Denmark, Portugal, the Netherlands, the UK and Austria. Roll out of the drug in Italy, Greece and France will follow as soon as pricing and reimbursement issues are resolved [412240], [412357]. By January 2001, the product was still under review and the company anticipated approval during the first half of 2001 in both the US and Europe [396802]. Launch in the US had originally been scheduled for 2000 [387293], [396802] and Japanese launch is planned for 2004 or 2005 [405915]. In January 2001, Amgen reported that the first pivotal trial of darbepoetin alfa in treating oncology patients with anemia was successful [396526], [396802]. The company anticipated a phase III trial in US patients in 2001 for the oncology indication 13977941. In the fourth quarter of 2000, darbepoetin alfa entered phase I trials in Japan. Japanese development of darbepoetin alfa was being conducted by Kirin Brewery (under the research code KRN-321) [396653]. In January 2001, Genesis Pharma licensed the rights to distribute, market and sell darbepoetin alfa for the treatment of anemia, in Greece and Cyprus [396437]. MegaPharm Ltd signed an agreement with Amgen in February 2001, granting MegaPharm certain exclusive rights to distribute, market and sell darbepoetin alfa in Israel [398897]. In February 2000, Merrill Lynch predicted that darbepoetin alfa sales will be in excess of $1.4 billion at maturity [355817]. In October 2000, darbepoetin alfa annual sales were predicted to hit $3 billion [387293]. PMID- 11892921 TI - Overview: pharmacogenomics of psychiatric disorders--separating the dreams from the realities. PMID- 11892922 TI - Rodent models of anxiety-like behaviors: are they predictive for compounds acting via non-benzodiazepine mechanisms? AB - An overview of the preclinical literature on the effects of the most studied clinically effective or putative non-benzodiazepine anxiolytics in existing animal models of anxiety-like behavior indicates that, with the exception of 5 HT1A agonists, these compounds display highly variable effects. Because these drugs have been shown (selective 5-HT reuptake inhibitors) or proposed (CCK(B) and CRF antagonists) to have a different spectrum of therapeutic activity in anxiety disorders than benzodiazepines, agents mainly used against generalized anxiety disorder, the screening of such compounds clearly requires the validation of new techniques that may model aspects of these conditions. PMID- 11892923 TI - Glutamate receptor ligands as anxiolytics. AB - The glutamatergic system has received considerable attention over recent years as a potential target for anxiolytic drugs. In spite of the pronounced anxiolytic like effects of competitive and non-competitive antagonists of NMDA receptors in animal models of anxiety, these substances can not be regarded as potential anxiolytic drugs, mainly due to their side-effect profiles (eg, ataxia, myorelaxation, impairment of learning and memory processes and psychotomimetic effects). Antagonists and partial agonists of the glycine, receptor inhibit function of the NMDA receptor complex and evoke in animals an anxiolytic-like response. Although data concerning anti-anxiety-like effects of glycine, receptor antagonists are not very promising, studies are underway to develop new, brain penetrating agents devoid of side effects. Further developments are necessary to more fully elucidate the possible involvement of AMPA/kainate receptors in anxiety. The recent discovery of metabotropic glutamate receptors, which modulate the function of the glutamatergic system, offers new hope for discovery of a new generation of anxiolytics. MPEP, a highly selective, brain penetrable, noncompetitive mGlu5 receptor antagonist, evokes anxiolytic-like effects in several animal models of anxiety, remaining remarkably free of side effects. LY 354740, a selective brain-penetrable group II mGlu receptor agonist, evokes marked anxiolytic-like effects in animal models of anxiety. LY-354740 causes mild sedation in mice, does not disturb motor coordination and has no potential to cause dependence. Therefore mGlu receptor ligands may become the anxiolytics of the future, free from the side effects characteristic of benzodiazepines. PMID- 11892924 TI - Gepirone. Organon. AB - Gepirone, a pyridinyl piperazine 5-HT1A receptor agonist, has been developed by Fabre-Kramer as an antidepressant. Bristol-Myers Squibb (BMS) outlicensed the compound to Fabre-Kramer in 1993 and is no longer involved in its development [337393]. In May 1998, NV Organon (a subsidiary of Akzo Nobel) licensed the rights to the drug product for further development and marketing from Fabre Kramer and, by October 1999, had submitted the drug for approval in the US [347133]. In December 2000, the company expected US and European launches in 2002 and 2003, respectively [402686]. Mechanism of action studies have demonstrated that gepirone, compared to buspirone, possesses a much greater selectivity for 5 HT1A receptors over dopamine D2 receptors. Long-term studies have shown that gepirone has a differential action at presynaptic (agonist) and post-synaptic (partial agonist) 5-HT1A receptors. However, further studies are still required to determine the relative contribution of pre- and post-synaptic 5-HT1A receptors to the therapeutic action of gepirone and related compounds. In March 2001, according to Schroder Salomon Smith Barney, Akzo Nobel targeted peak sales of Euro 300 million for gepirone [409013]. This amount was reiterated in an April 2001 report by HSBC Securities, which stated that gepirone was expected to achieve this figure in 2009 or 2010 [409014]. PMID- 11892925 TI - Overview: changing times: developing cancer drugs in genomeland. PMID- 11892926 TI - Mutant p53: the loaded gun. AB - Alterations in the p53 gene are the most common genetic defects found in tumors so far. Taking into account that p53 is a powerful inducer of cell death it is not surprising that the abolition of its function occurs almost universally during tumor development. There are several features of p53 inactivation in tumors which are quite unique. Firstly, mutations occur at high frequency in the p53 gene, ie, around 50% of human tumors carry p53 mutations. Secondly, mutations are largely of the same type, ie, 87% of them are point missense mutations resulting in a substitution of one amino acid residue. Thirdly, the majority of mutations occur in the DNA binding domain of p53. Finally, mutant p53 proteins accumulate at high levels in tumor cells. Can we take advantage of p53 mutations in tumor cells to selectively kill them? Is this the Achilles heel of tumors that can be exploited for novel non-toxic anticancer therapy? In this review the possible approaches toward reactivation of mutant p53 in tumors will be discussed. PMID- 11892927 TI - Su-6668. SUGEN. AB - SUGEN is developing SU-6668, a tyrosine kinase inhibitor that inhibits three distinct growth factor receptor targets, for the potential treatment of cancer [304530]. The compound is in phase I trials in the UK and US [321260], [374505]. A report in January 2001 stated that phase I/II trials for hematological and solid tumors were expected to commence shortly thereafter [395657]. In May 2001, phase I data from a dose-escalation study conducted at UCLA were presented at the 37th ASCO meeting in San Francisco, CA. By that time, 74 patients had been enrolled in this study which aimed to determine the toxicities of SU-6668 when delivered to fed and fasting patients. SU-6668 was administered orally either once or twice-daily at doses of 100 to 2400 mg/m2 to patients diagnosed as having advanced malignancies. Accrual in phase I is continuing to define the toxicities of doses > 200 mg/m2 twice-daily [409984], [411418]. In December 1998, SUGEN filed an IND with the FDA for the clinical testing of this compound with oral and iv formulations [310237]. In November 1998, SUGEN entered into a collaboration with the Cancer Research Campaign (CRC) to conduct a phase I trial of SU-6668 at the Royal Marsden Hospital, UK using an iv formulation [304530]. After the first six patients had been treated, the trial was halted owing to problems with the iv formulation. Some toxicities, including nausea, vomiting, fatigue and tumor pain were observed [413538]. No licensing agreement as involved between the CRC and SUGEN for this trial [408572]. PMID- 11892928 TI - Flavopiridol. National Cancer Institute. AB - Flavopiridol is a synthetic flavonoid inhibitor of cyclin-dependent kinases, which is under development by Aventis Pharma (formerly Hoechst Marion Roussel) and the National Cancer Institute (NCI) for the potential treatment of cancer and proliferative disorders. By May 2001, the product was in phase IIa trials and had achieved proof-of-concept in phase I/IIa trials as a monotherapy. At this time, Aventis expected a global submission to take place in 2003 [409257]. By July 1999, the compound had entered phase II trials for gastric cancer and leukemia, and phase I/II trials for esophageal tumor and non-small cell lung cancer (NSCLC) [277372], [325929], [331850]. Phase II trials for colon and renal cancer [411684], [411769] and phase I trials for prostate cancer [279466] have also been reported. Analysts Merrill Lynch predicted in September and November 2000 that the product would be launched by 2003, with sales of EUR 50 million in that year, rising to EUR 100 million in 2004 [383742], [391426]. In April 1999, ABN Amro predicted annual sales of DM 100 million in 2002 [328676]. PMID- 11892929 TI - Combinatorial biosynthesis in microorganisms as a route to new antimicrobial, antitumor and neuroregenerative drugs. AB - Combinatorial biosynthesis utilizes the genes of biosynthetic pathways that produce microbial products to create novel chemical structures. The engineering of mondular polyketide synthase (PKS) genes has been the major focus of this effort and has led to the production of analogs of macrolide antibiotics like the erythromycins and their derived ketolides, and of the immunosuppressive macrolide FK-520 (Fujisawa Pharmaceutical Co Ltd). Approaches to making analogs of the promising antitumor compounds known as epothilones are also being explored. Lead compounds for further study have resulted and routes to analogs of other pharmacologically important compounds have been established. To facilitate this work, many new tools for manipulating and studying the multifunctional PKSs have been developed including the development of Escherichia coli as a PKS expression last. These developments have resulted in faster ways of engineering PKS to produce new compounds for the development of chemotherapeutic agents from natural products. PMID- 11892930 TI - Telithromycin. Aventis Pharma. AB - The ketolide telithromycin (HMR-3647; Ketek), a derivative of clarithromycin, has been launched by Aventis Pharma (formerly Hoechst Marion Roussel) for the treatment of respiratory tract infections with gram-positive or gram-negative cocci, Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, intracellular pathogens, atypical microorganisms, toxoplasma or anaerobic bacteria. By May 2001, filings in the US and EU had been completed and a filing in Japan was expected to take place in the fourth quarter of 2001. In July 2001, telithromycin was granted marketing authorization by the EC for the treatment of community-acquired respiratory tract infections, including those caused by bacteria resistant to commonly used antibiotics. In October 2001, the product was launched in Germany. In March 2000, telithromycin was submitted to the US FDA and the EMEA, under the EU centralized approval procedure, for approval for the treatment of community-acquired pneumonia (CAP), acute sinusitis, acute exacerbations of chronic bronchitis and tonsillitis/pharyngitis. The company had expected to launch the product in early 2001. The CPMP issued a positive opinion for all four indications on April 23 2001. In September 2001, the company indicated that it expected the product to be launched in Japan in 2002. The FDA's Anti-infectives Advisory Committee was due to review telithromycin for all the submitted indications on January 29 2001; however, this was postponed. This postponement was thought to be at Aventis' request in order to discuss the potential for a resistant pneumococcal infection labeling which would boost product sales. The revised date for the meeting was April 26 2001, at which the Anti-Infective Drugs Advisory Committee of the FDA recommended approval of telithromycin for the treatment of CAP in patients 18 years of age or older. The committee failed to recommend approval for the use of the drug for the remaining three indications for which it was filed, citing concerns over potential cardiovascular risk and liver toxicity; at this time, the company was in active discussions with the FDA regarding approval of the remaining three indications. An approvable letter for CAP, acute bacterial exacerbations of chronic bronchitis and acute bacterial sinusitis was received by the company in June 2001; Aventis also received a non-approvable letter for the treatment of tonsillitis/pharyngitis at this time. In April 1999, ABN Amro predicted annual sales of DM 50 million in 2001, rising to DM 100 million in 2002. In February 1999, Lehman Brothers estimated a 70% probability that this ketolide would come to market. The analysts also estimated a launch date of 2001, with peak sales of US $700 million in 2009. Analysts Merrill Lynch predicted in September 200, that the product would be launched by 2001, with sales of euro 50 million in that year, rising to euro 284 million in 2004. Deutsche Bank predicted in August 2001, that sales of the product would reach euro 5 million in 2001, rising to euro 300 million in 2005. Analysts at Merrill Lynch predicted in November 2001, that the product would be resubmitted in the US in mid-2002, and would make sales of US $5 million in 2001, rising to US $250 million in 2004. PMID- 11892931 TI - HMR-3562. Aventis Pharma. AB - Aventis Pharma (formerly Hoechst Marion Roussel) is investigating the ketolides HMR-3562 and HMR-3787 as potential antibacterial agents. The compounds belong to a series of 2-fluoroketolides. They have exhibited potency against a range of gram-positive bacteria and other respiratory tract pathogens including Haemophilus influenzae in vitro and in vivo. PMID- 11892932 TI - T-3811. Toyama/Bristol-Myers Squibb. AB - Toyama and Bristol-Myers Squibb (BMS) are developing the des-F(6)-quinolone T 3811 (BMS-284756) as a potential treatment for bacterial infections, including respiratory and urinary tract infections, and skin and soft tissue infections. The drug is in phase II trials in the US and Europe, and an injectable formulation began phase I trials in the US in April 2000. Phase I trials commenced in Japan in September 1999. Phase III trials were expected to begin outside Japan in 2000. T-3811 is expected to be administered as an oral or injectable once-a-day formulation. Preclinical studies suggests it has a better side-effect profile than commonly associated with other quinolones on the market. Bristol-Myers Squibb has acquired worldwide development and marketing rights, with the exception of Japan, South Korea, Taiwan and China. In December 2001, Morgan Stanley predicted that T-3811 could have peak sales potential exceeding $500 million if the positive clinical data released at this year's ICAAC continued, and that BMS was on track for filing in 3Q02. PMID- 11892933 TI - Targeting the chemokine system for multiple sclerosis treatment. AB - Multiple sclerosis (MS) is the most common demyelinating disorder of the human central nervous system (CNS). The typical pathological hallmark of active MS is the presence of inflammatory foci disseminated in the CNS. It is believed that the composition of inflammatory infiltrates is determined in part by the spectrum of chemokines produced in a focus of inflammation. Numerous studies suggest chemokine involvement in MS pathogenesis. Interfering with chemokine-chemokine receptor interactions may potentially lead to prevention and/or amelioration of CNS inflammatory processes. Initial studies to obtain 'proof-of-principle' used neutralizing antibodies in small animal models of MS. The subsequent generation of chemokine receptor inhibitors were modified chemokine peptides. At present, the development of small molecule antagonists to chemokine receptors is the dominant approach. Current evidence suggests that chemokines and their receptors are promising targets for effective treatment of MS and other CNS inflammatory and autoimmune diseases. PMID- 11892934 TI - Endothelium as a pharmacological target. AB - Over the last few years, the increasing knowledge of the endothelium has highlighted its integral role in a number of pathologies. Endothelial cells are pivotally involved in the recruitment and adhesion of leukocytes and platelets, and they express adhesion molecules and growth factors. This review highlights the recent advances made in the understanding of the endothelium and discusses the endothelium as a potential target in a variety of diseases, including cardiovascular diseases, cancer and inflammatory diseases. PMID- 11892935 TI - Etanercept. Immunex. AB - Immunex has developed and launched etanercept, a soluble TNF receptor (TNFR) fusion protein, for the treatment of rheumatoid arthritis (RA). It has also been developed for various TNF-mediated conditions such as congestive heart failure, endometriosis and multiple sclerosis. Etanercept has been launched as a second line agent in the US for the treatment of moderate-to-severe RA and can be used in conjunction with methotrexate in patients unresponsive to methotrexate alone. It is also available in the EU. In 2000, it was in phase III trials for psoriatic arthritis and an NDA filing for this indication was expected for the first half of 2001. In July 2001, the sBLA was filed, and in September 2001, the FDA granted the sBLA Priority Review status. As of January 2001, etanercept was in phase III trials for congestive heart failure, with sNDA filing expected in 2002; however, by March 2001, these had been halted, as it did not appear that statistical significance would be reached for the efficacy endpoints. Further data analysis was being undertaken at this time, before a final decision was taken. In April 2001, Merrill Lynch reported that development for this indication was to be halted. Sales for the drugs first full quarter on the market in 1999 were US $59.7 million. By November 1999 the drug had made sales of US $500 million; Immunex expected the drug to generate over US $2 billion in annual sales by 2004. In September 2000, Merrill Lynch reported that if sales of the drug continued at the present rate then it is likely that demand would temporarily outstrip supply in 2001. Resolution of the supply issue was expected by 2002. Also in September 2000, Merrill Lynch lowered their estimate of sales in 2001 from US $1 billion to $927 million. In the long-term, Merrill Lynch believed that the drug has the potential to exceed US $5 billion in sales in the US. In April 2001, Merrill Lynch predicted that etanercept prescribed for RA would generate sales of US $71 in 2002 rising to US $600 million in 2005. In October 2001, Morgani Stanley reported that Enbrel continues to be the primary source of revenue of Immunex (US $198.1 million). It was also reported that if launched for CHF, an estimated peak year revenue was likely to be US $500 million. The company maintains a website containing additional information about etanercept at http://www.enbrelinfo.com. PMID- 11892936 TI - An update of neuroprotectants in clinical development for acute stroke. AB - Neuroprotectants are drugs designed to treat stroke by preserving ischemic neurons in the penumbra. Despite numerous studies over the past ten years, no such drug has yet shown clinical efficacy. This article reviews those trials completed since 1999, including assessments of drugs that modify receptors or ion flow, block leukocyte adhesion receptors or stabilize membranes. In addition, ongoing trials and early trials using novel mechanisms of action, such as hypothermia and antioxidants, are discussed. PMID- 11892937 TI - UK-315716/UK-240455. Pfizer. AB - Pfizer is developing the atropoisomeric quinoxalinediones, UK-315716 and UK 240255, from a series of NMDA/glycine antagonists for the potential treatment of stroke. Both are compounds are undergoing clinical trials. UK-315716 has improved aqueous solubility and in vivo efficacy over UK-240455, and a lower projected clinical dose of 200 mg compared to 500 mg with UK-240455. PMID- 11892938 TI - KB-R7943. Kanebo. AB - Kanebo is investigating KB-R7943, a Na+/Ca2+ ion exchange inhibitor, for the potential treatment of ischemia and reperfusion injury. It inhibited the outward Na+/Ca2+ exchange current (iNCX) more potently than the inward current under unidirectional flow conditions; however, inward and outward current were inhibited equally under bidirectional conditions. The drug was a competitive inhibitor to external calcium, and the inhibition was reversible with a recovery t1/2 of about 30 s. The mammalian Na+/Ca2+ exchanger forms a multigene family of homologous proteins comprising three isoforms, NCX1, NCX2 and NCX3. By examining chimeric constructs between NCX1 and NCX3 expressed in CCL39 cells, it has been demonstrated that it is the conserved internal repeat regions (alpha-1 and alpha 2) of the exchanger that are critical for the drug's action. PMID- 11892939 TI - Overview: a molecule for all occasions. PMID- 11892940 TI - Overview: pro-inflammatory cytokines in cerebrovascular ischemia. PMID- 11892941 TI - Cladribine. Ortho Biotech Inc. AB - Cladribine, an adenosine deaminase inhibitor, has been developed and launched by Ortho Biotech in collaboration with The Scripps Research Institute for the treatment of several neoplasms, including acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, cutaneos T-cell lymphoma, hairy-cell leukemia and non-Hodgkin's lymphoma. It was first launched in the US in February 1993. Ortho Biotech and The Scripps Research Institute have since been developing the compound for its potential use in multiple sclerosis (MS). In 1997, Ortho filed air NDA in the US for the use of cladribine in the treatment of relapsing-remitting and secondary progressive MS. An FDA drug advisory committee was planning to meet in January 1999 to discuss the NDA. However, Ortho cancelled the meeting. Following an FDA inspection during December 1998 and January 1999, the Scripps Clinic received a warning letter from the FDA in April 1999 regarding violations in the clinical studies of cladribine for MS, and Ortho withdrew the NDA after concluding that further clinical studies would be necessary. Cladribine has been known since the 1960s as an intermediate for the synthesis of 2 deoxynucleotides and its potential for the treatment of leukemia was disclosed in 1984. The Scripps Research Institute and the Johnson & Johnson group hold several patents claiming preparation methods (US 05208327), and additional indications, such as multiple sclerosis (WO-09316706) and rheumatoid arthritis (US-05310732). The associated patent, WO-09323508, is the only one among those patents that claims the use of unmodified cladribine for the treatment of leukemia, but it focuses particularly on a specific form of the disease, chronic myelogenous leukemia. Analysts at UBS Warburg predicted in October 2001, that the product would make US sales of $50 million in 2004 for its MS indication. PMID- 11892942 TI - Citicoline. Ferrer Internacional. AB - Citicoline was originally developed and launched by Ferrer for the treatment of stroke, and is now also being investigated for the potential treatment of Alzheimer's disease (AD). In the US, the compound is being developed by Interneuron for the treatment of stroke. A US launch had been rescheduled for 2002 although a decision on future US development of citicoline was intended to be made in conjunction with Takeda, Interneuron's US licensee. Takeda had decided not to pursue development by December 2000 and was in negotiations with Interneuron for another product candidate. Interneuron stated at this time that it would explore other partnership opportunities for citicoline. In 1993, Interneuron licensed exclusive marketing and manufacturing rights to citicoline in the US and Canada from Ferrer. By September 1997, a patent application had been filed worldwide by Interneuron for the use of citicoline in the reduction of cerebral infarct volume, and in September 1998, US-05801160 was issued for citicoline relating to the protection of brain tissue from cerebral infarction following ischemic stroke. In December 1999, US rights to the commercialization of citicoline were licensed to Takeda. PMID- 11892943 TI - Losigamone. Dr Willmar Schwabe. AB - Losigamone is a potential antiepileptic under development by Schwabe, which is currently in phase III clinical trials. By August 2000, Schwabe was seeking suitable partners to collaborate in the completion of the phase III trials and further development. The exact mode of action of losigamone is unclear but it does not involve specific binding of GABA, flunitrazepam or t-butyl bicyclophosphorothionate (TBPS) to their receptors. Data concerning the interaction of losigamone with GABA-A receptor channels, however, are inconsistent; it does not significantly modify the GABAergic inhibitory postsynaptic potentials (IPSPs) in hippocampal slices, although it potentiates GABA-induced chloride influx in primary spinal cord neuron cultures. Another suggested possible mechanism of action for losigamone is K+ channel activation. PMID- 11892944 TI - Overview: synthetic gene delivery: another quest for the Holy Grail? PMID- 11892946 TI - INGN-201. Introgen Therapeutics. AB - In April 2001, Aventis and Introgen signed a letter of intent to restructure their collaboration arrangement, giving Introgen responsibility for the worldwide development of all p53 programs under the existing collaboration and obtain exclusive worldwide commercial rights to p53-based gene therapy products, including INGN-201. In February 2001, Introgen was awarded US-06194191 for the commercial production of adenovirus vectors. In November 2000, the company and the University of Texas System were issued with US-06143290, entitled 'Recombinant p53 adenovirus methods and compositions', further solidifying the company's current p53 patent portfolio. In June 2000, the University of Texas System was awarded US-06069134, entitled, 'Methods and compositions comprising DNA damaging agents and p53.' This is the second US patent to be issued that is equivalent to WO-09528948. In May 1998, US-05747469, entitled 'Methods and compositions comprising DNA damaging agents and p53', was awarded to the Board of Regents of the University of Texas. This patent covers the use of the p53 gene in combination with chemotherapeutic agents, radiation therapies or other agents, which damage the DNA of cancer cells. It is one of several intellectual properties licensed to Introgen through an agreement with the MD Anderson Cancer Center. PMID- 11892945 TI - ONYX-015. Onyx Pharmaceuticals. AB - ONYX-015 (CI-1042), an adenovirus modified selectively to replicate in and kill cells that harbor p53 mutations, is under development by Onyx Pharmaceuticals for the potential treatment of various solid tumors, including head and neck, gastrointestinal and pancreatic tumors. It is a recombinant adenovirus that carries a loss-of-function mutation at the E1B locus, the product of which is a 55 kDa protein that binds to and inactivates the p53 tumor suppressor protein. Wild-type adenoviruses must disable this gene before viral replication can occur. This, the ONYX-015 adenovirus will leave normal cells unaffected. Mutations in the p53 tumor suppressor gene are the most common type of genetic abnormality in cancer, occurring in more than half of all major cancer types. Thus, these cells are susceptible to the virus, which will readily replicate and cause cell death. ONYX-015 is in ongoing phase III trials for the treatment of recurrent head and neck cancer, phase II trials for colorectal, ovary, pancreas and mouth tumors, and phase I trials for digestive disease, esophagus and liver tumors. Onyx Pharmaceuticals was granted US-05677178 covering methods for the treatment of p53 related cancers in October 1997. The patent specifically covers the use of modified adenoviruses and other DNA viruses, which lack viral proteins that bind to p53, for the treatment of cancer patients whose tumors lack p53 function. The USPTO awarded Onyx Pharmaceuticals US-05846945 in December 1998, covering methods for treating cancer using replicating adenoviral therapy in combination with chemotherapy. In April 1999, the company also received EP-094910177.8 covering the technology in Europe. PMID- 11892947 TI - Neurocognitive developmental disorders: a real challenge for developmental neuropsychology. PMID- 11892948 TI - Atypical brain development: a conceptual framework for understanding developmental learning disabilities. AB - This article presents ideas that are, in part, a response to the ambiguity in the neurological research on learning disorders, the growing awareness that developmental disabilities are typically nonspecific and heterogeneous, and the growing scientific literature showing that comorbidity of symptoms and syndromes is the rule rather than the exception. This article proposes the term atypical brain development (ABD) as a unifying concept to assist researchers and educators trying to come to terms with these dilemmas. ABD is meant to serve as an integrative concept of etiology, the expression of which is variable within and across individuals. ABD does not itself represent a specific disorder or disease. It is a term that can be used to address the full range of developmental disorders that are found to be overlapping much of the time in any sample of children. Although similar in spirit to the older term of minimal brain dysfunction (MBD), in that it closely links neurology with behavioral difficulties, ABD as proposed here differs in several ways. In support of the ABD conceptual framework, first, we consider the ABD concept in terms of its superiority to the older notion of MBD. Second, we provide a brief review of the burgeoning literature on the overlap of the various developmental disabilities. Third, we review some of the scientific literature that supports the ABD concept. Our sole purpose in proposing this concept is to initiate dialogue and debate on several critical issues across a wide variety disciplines. Hence, this article is not intended to be a definitive statement of a rigid perspective. It reflects neither a nonmalleable philosophical position, nor any type of condemnation of other perspectives. It does, however, reflect a data-based and philosophical trend visible in the field of learning disabilities, as well as the broader area of childhood developmental disorders. PMID- 11892949 TI - Genetic and environmental influences on orthographic and phonological skills in children with reading disabilities. AB - Data from identical and fraternal twins were analyzed to estimate the proportions of genetic and environmental influences on group deficits in accuracy and, when available, speed for printed word recognition and for related skills in phonological decoding (PD), orthographic coding (OC), and phoneme awareness (PA). In addition, bivariate genetic analyses were employed to estimate the degree of common genetic influence on group deficits across these different reading and language skills. About half of the group deficits in each of the skills were due to genetic influences, and the genetic origins were largely shared among the measures (r(g) = .53 - .99), except for those between OC and PA (r(g) = .28 - .39). Implications of the results are discussed for models of reading disability and remediation. PMID- 11892950 TI - The sensory basis of reading problems. AB - Learning to read is much more difficult than learning to speak. Most children teach themselves to speak with little or no difficulty. Yet a few years later when they come to learn to read they have to be taught how to do it; they do not pick up reading by themselves. This is because we speak in words and syllables, but we write in phonemes. Syllables do not naturally break down into the sounds of letters and letter units (i.e., phonemes) because these do not correspond to physiologically distinct articulatory gestures (Liberman, Shankweiler, & Studdert Kennedy, 1967). Alphabetic writing was only invented when people realized that syllables could be artificially divided into smaller acoustically distinguishable phonemes that could be represented by a small number of letters. But these distinctions are arbitrary cultural artifacts, and their mastery was originally confined to a select social class. And until about 100 years ago it did not matter much if the majority of people could not read; the acquisition of reading probably had no serious disadvantages. Reading requires the integration of at least two kinds of analysis (Castles & Coltheart, 1993; Ellis, 1984; Manis, Seidenberg, Doi, McBride-Chang, & Petersen, 1996; Morton, 1969; Seidenburg, 1993). First, the visual form of words, the shape of letters, their order in words, and common spelling patterns, which is termed their orthography, has to be processed visually. Their orthography yields the meaning of familiar words very rapidly without needing to sound them out. But for unfamiliar words, and all words are fairly unfamiliar to the beginning reader, the letters have to be translated into the speech sounds (i.e., phonemes) that they stand for, and then those sounds have to be melded together in inner speech to yield the word and its meaning. Reading exclusively by the phonological route is more time consuming than if words can be accessed directly without requiring phonological mediation. PMID- 11892951 TI - Developmental pathways of children with and without familial risk for dyslexia during the first years of life. AB - Comparisons of the developmental pathways of the first 5 years of life for children with (N = 107) and without (N = 93) familial risk for dyslexia observed in the Jyvaskyla Longitudinal study of Dyslexia are reviewed. The earliest differences between groups were found at the ages of a few days and at 6 months in brain event-related potential responses to speech sounds and in head-turn responses (at 6 months), conditioned to reflect categorical perception of speech stimuli. The development of vocalization and motor behavior, based on parental report of the time of reaching significant milestones, or the growth of vocabulary (using the MacArthur Communicative Development Inventories) failed to reveal differences before age 2. Similarly, no group differences were found in cognitive and language development assessed by the Bayley Scales of Infant Development and the Reynell Developmental Language Scales before age 2.5. The earliest language measure that showed lower scores among the at-risk group was maximum sentence length at age 2. Early gross motor development had higher correlation to later language skills among the at-risk group rather than the control children. The most consistent predictor of differential development between groups was the onset of talking. Children who were identified as late talkers at age 2 were still delayed at the age 3.5 in most features of language related skills-but only if they belonged to the group at familial risk for dyslexia. Several phonological and naming measures known to correlate with reading from preschool age differentiated the groups consistently from age 3.5. Our findings imply that a marked proportion of children at familial risk for dyslexia follow atypical neurodevelopmental paths. The signs listed previously comprise a pool of candidates for early predictors and precursors of dyslexia, which await validation. PMID- 11892952 TI - What framework should we use for understanding developmental disorders? AB - The neuropsychology of dyslexia has made great strides in the last decade. In particular, a consensus views dyslexia as a developmental disorder with a basis in the brain and in the genes, where the interaction of genetic and environmental factors is taken for granted. However, problems in defining the phenotype continue to bedevil research. The main conceptual problems can be expressed in three main questions: (a) Is dyslexia based on a specific brain abnormality or is it merely part of a continuum of atypical brain development? (b) When can we speak of comorbidity? (c) Why does so much individual variability occur? These questions can be tackled in a common framework that takes into account simultaneously three levels: the biological, the cognitive, and the behavioral. PMID- 11892953 TI - From the Centers for Disease Control and Prevention. Tuberculosis morbidity among U.S.-born and foreign-born populations--United States, 2000. AB - In collaboration with all state health departments, CDC conducts public health surveillance for tuberculosis (TB). This report summarizes data from the national TB surveillance system for 2000 and compares them with data from 1992-1999. During 2000, a total of 16,377 cases (5.8 cases per 100,000 population) of TB were reported to CDC from the 50 states and the District of Columbia (DC), representing a 7% decrease from 1999 and a 39% decrease from 1992, when the number of cases and case rate most recently peaked in the United States. However, the case rate among foreign-born persons remains at least seven times higher than among U.S.-born persons. To address the high rate, CDC is collaborating with public health partners to implement TB control initiatives among recent international arrivals and residents along the border between the United States and Mexico and to strengthen TB programs in countries with a high incidence of TB disease. PMID- 11892955 TI - From the Centers for Disease Control and Prevention. Progress toward elimination of perinatal HIV infection--Michigan, 1993-2000. AB - In 1994, the U.S. Public Health Service (PHS) issued guidelines for maternal and neonatal zidovudine (ZDV) use to reduce perinatal human immunodeficiency virus (HIV) transmission. These guidelines recommend maternal ZDV use during the second and third trimesters of pregnancy and during labor and delivery (L&D) and administration of ZDV to the neonate for the first 6 weeks of life. In 2001, PHS updated 1995 guidelines for routine HIV counseling and voluntary testing of pregnant women. The Michigan Department of Community Health (MDCH) requires reporting of all children who are perinatally exposed to HIV and follows up these children to monitor their infection status and record demographic, clinical, and laboratory characteristics of infected children. The reporting of perinatally HIV exposed children enables MDCH to monitor the effectiveness of public health efforts to prevent perinatal HIV transmission and assists the targeting of prevention programs and activities. This report summarizes surveillance data collected through December 31, 2001, on children born to HIV-infected women in Michigan during 1993-2000. The report highlights rapid adoption of PHS guidelines that resulted in the reduction of perinatally acquired HIV infection to historically low levels in Michigan. Improving levels of prenatal care (PNC) for HIV-infected pregnant women, especially substance users, and routine HIV counseling and voluntary testing for all pregnant women are needed to further reduce perinatal HIV infection. PMID- 11892956 TI - From the Centers of Disease Control and Prevention. Use of assisted reproductive technology--United States, 1996 and 1998. AB - Since 1983, when the first infant was conceived from in vitro fertilization (IVF) in the United States, the use of IVF and related procedures (assisted reproductive technology [ART]) has increased substantially. In 1998, an estimated 0.7% of the 3.9 million births were the result of ART. ART patients are more likely to deliver multiple infants than women who conceive without treatment, and these multiple-infant births are associated with increased risks for pregnancy complications, premature delivery, low birth-weight infants, and long-term disability among surviving infants. This report examines state-specific use of ART in 1996 and 1998 and provides data on ART live-born and multiple-infant birth rates in 1998. Findings indicate that the use of ART is increasing in most states and that more than half the infants born as a result of these procedures are multiple births. These high-risk births contribute disproportionately to health care costs and might negatively affect maternal and child health outcomes, particularly in states where large numbers of ART procedures are performed. PMID- 11892957 TI - Defining parkinsonism in the Canadian Study of Health and Aging. AB - This study sought an operational definition of parkinsonism in elderly people (n = 2,914) who underwent a clinical examination in the Canadian Study of Health and Aging (CSHA). Parkinsonism was defined as having two of the following features: (1) bradykinesia of face or limbs, (2) resting tremor, (3) rigidity, and (4) abnormality of gait and posture. The association of parkinsonism with other parkinsonian-related features (prior diagnosis of Parkinson's disease, use of drugs with extrapyramidal side effects, and use of antiparkinsonian medications) and variables not expected to be related to parkinsonism (stroke and Hachinski score > 5) was determined. Parkinsonism was identified in 337 people (11.6%). It was significantly more likely with other parkinsonian-related characteristics, and was not associated with a history of stroke, but was slightly higher among those subjects with a Hachinski score > 5. Posture and gait abnormalities were significantly associated with other parkinsonian-related variables, but were also more common among subjects with stroke-related features. When the gait and posture disturbance category was excluded as a parkinsonian sign, the narrower definition was more specific but less sensitive in detecting cases with a clinical diagnosis of Parkinson's disease. Despite limitations, the approach presented in this article is a valid method to operationalize parkinsonism from the dataset. PMID- 11892958 TI - Assessing hypertension in the Canadian Study of Health and Aging. AB - We investigated the self-report hypertension variables in the CSHA, recorded in the screening questionnaire and the Self-Administered Risk Factor (SARF) questionnaire. The two questions showed high agreement (phi coefficient 0.83). Each was modestly but significantly associated with other simultaneous reports of heart disease and stroke, and with subsequent mortality. Only the SARF asked questions about treatment; controlling for treatment effects, five-year survival was longest among those with no hypertension and no treatment (mean survival time 1,645 days; 95% CI 1,632 to 1,658), and shortest for those with no reported hypertension who were receiving "antihypertensive" medications presumably prescribed for other cardiovascular disease (mean survival time 1,496 days; 95% CI 1,457 to 1,535). The SARF questions incorporating high blood pressure and treatment appear preferable to assess the risks associated with hypertension. PMID- 11892959 TI - Development and validation of an indicator of support for community-residing older Canadians. AB - Lack of social support is an important risk factor for disability, psychiatric illness, cognitive impairment, institutionalization, and mortality. Social networks are also important for the caregiving and emotional support that elderly people need to allow them to function well in the community. This article details the development and validation of an index of the instrumental support available to older community residents in the Canadian Study of Health and Aging (CSHA). Preliminary item review, internal consistency, and exploratory factor analysis were carried out on a random half of the sample. The second half of the sample was used for cross-validation; internal consistency, exploratory factor analysis, and item response theory analysis were carried out. The final scale had six items; alpha internal consistency was 0.76 and IRT reliability was 0.85. A one factor solution was most easily interpretable. IRT analyses showed that the scale was homogeneous and that most items were highly discriminating. The instrumental support scale also had a high correlation with size of social network; it was related to marital status and gender, and predicted institutionalization between the two phases of the study. PMID- 11892960 TI - Use of the chronic disease score to measure comorbidity in the Canadian Study of Health and Aging. AB - Most older adults have multiple chronic diseases. Consideration of these conditions can improve the performance of statistical models in epidemiological analyses. The Chronic Disease Score (CDS) is a measure of comorbidity derived from medication usage, which may have some advantages over measures derived from other sources. The calculation of the CDS from data contained in the Canadian Study of Health and Aging (CSHA) is described. This measure can be used to estimate comorbidity within the CSHA database. PMID- 11892961 TI - Effects of screening errors and differential mortality on the estimation of the incidence of dementia in the Canadian Study of Health and Aging. AB - The Canadian Study of Health and Aging produced an estimate of the incidence of dementia among elderly Canadians by following up, after 5 years, the undemented found in an initial prevalence survey. Initial and follow-up estimates could be biased by false-negative error in the screening tool used for subjects living in the community, and by erroneous classification of subjects who died in the interim. Here, we use a deterministic model to quantify those possible biases. We conclude that, using the estimates of the errors from control samples, the incidence among community subjects would be overestimated by 15%, and the incidence among the institutional subjects would be underestimated by 37%. The overall incidence would be underestimated by 14%. Most of the bias can be attributed to inaccuracies in the classification of deaths. PMID- 11892962 TI - Linkage of the Canadian Study of Health and Aging to provincial administrative health care databases in Nova Scotia. AB - The Canadian Study of Health and Aging (CSHA) was a cohort study that included 528 Nova Scotian community-dwelling participants. Linkage of CSHA and provincial Medical Services Insurance (MSI) data enabled examination of health care utilization in this subsample. This article discusses methodological and ethical issues of database linkage and explores variation in the use of health services by demographic variables and health status. Utilization over 24 months following baseline was extracted from MSI's physician claims, hospital discharge abstracts, and Pharmacare claims databases. Twenty-nine subjects refused consent for access to their MSI file; health card numbers for three others could not be retrieved. A significant difference in healthcare use by age and self-rated health was revealed. Linkage of population-based data with provincial administrative health care databases has the potential to guide health care planning and resource allocation. This process must include steps to ensure protection of confidentiality. Standard practices for linkage consent and routine follow-up should be adopted. The Canadian Study of Health and Aging (CSHA) began in 1991-92 to explore dementia, frailty, and adverse health outcomes (Canadian Study of Health and Aging Working Group, 1994). The original CSHA proposal included linkage to provincial administrative health care databases by the individual CSHA study centers to enhance information on health care utilization and outcomes of study participants. In Nova Scotia, the Medical Services Insurance (MSI) administration, which drew the sampling frame for the original CSHA, did not retain the list of corresponding health card numbers. Furthermore, consent for this access was not asked of participants at the time of the first interview. The objectives of this study reported here were to examine the feasibility and ethical considerations of linking data from the CSHA to MSI utilization data, and to explore variation in health services use by demographic and health status characteristics in the Nova Scotia community cohort. PMID- 11892963 TI - Disability and frailty among elderly Canadians: a comparison of six surveys. AB - The first wave of the Canadian Study of Health and Aging (CSHA) constituted a large health survey of a representative sample of elderly Canadians. Other Canadian surveys from the same era provided equivalent figures, and the present report compares the results of 6 surveys on a variety of health indicators. Agreement was close on self-reported chronic health conditions, adequate for several indicators of functional limitation, but was lower for overall self ratings of the impact of health problems on day-to-day life. Using the CSHA data to compare alternative operational definitions of frailty, a definition based on ADL limitations appeared to offer an underestimate; addition of IADL questions or cognitive limitations provided figures that appeared more plausible. Survey estimates of chronic health conditions appear consistent, as are estimates of certain ADL disabilities. Care must be taken with interpreting more subjective reports, while prevalence of frailty varies considerably according to the definition used. PMID- 11892964 TI - Estimating the prevalence of dementia in elderly people: a comparison of the Canadian Study of Health and Aging and National Population Health Survey approaches. AB - The Canadian Study of Health and Aging (CSHA) and the National Population Health Survey (NPHS) collected data on the prevalence of dementia in differing fashions. The CSHA used a two-stage method with objective testing and expert judgment, and the NPHS used self-report and proxy data. The present report compares estimates of prevalence and the methodology for ascertainment in the two surveys. The more detailed approach of the CSHA offers the more valid means of estimating prevalence and providing data on subtypes, and can be used in naturalhistory studies. TheNPHSmeasures, including a self/proxy report of diagnosed dementia and a derived cognitive measure, are not sufficiently valid for useful inferences to be made. However, the NPHS method can be improved through supplementation with data on functional disability, providing age group-specific point estimates closer to the CSHA's estimates of cognitive impairment and dementia from the community sample. Future waves of the NPHS may wish to include objective cognitive function measures as a cost-efficient and more accurate method of estimating the prevalence of the dementia syndrome without attempting to estimate the prevalence of particular causes of that syndrome. PMID- 11892965 TI - Reliability and validity of questions about exercise in the Canadian Study of Health and Aging. AB - Regular exercise in elderly people has beneficial health effects. We examined exercise frequency and intensity from the Canadian Study of Health and Aging Risk Factor Questionnaire (RFQ). The reliability and validity of these two questions individually, and when combined to form a scale, are reported. Agreement between the self-administered RFQ and an interviewer-administered Add-on Study was examined using intraclass correlations, which were 0.80 for frequency (95% CI 0.77-0.82, p < .001) and 0.75 for intensity (95% CI 0.71-0.78, p = .012). Individuals reporting high levels of exercise frequency, intensity, and a combination of the two showed a smaller proportion of adverse health markers than those reporting no regular exercise. Predictive validity assessed by Cox proportional hazards modeling of mortality showed that the high and moderate levels of frequency, intensity, and combined exercise groups differed significantly (all p < .001) from the no exercise group. We have found that these exercise questions, though simple, appear reliable and valid. The finding that even comparatively crude exercise questions can demonstrate an important relationship to death suggests that the signal for exercise is a strong one, and future studies should seek to better examine mechanisms by which exercise benefit is conferred. PMID- 11892966 TI - Cognitive impairment, no dementia: concepts and issues. AB - This article reviews the concept of mild cognitive impairment in groups of people whose cognitive impairment does not warrant a diagnosis of dementia (cognitive impairment, no dementia; CIND). Problems with the application of existing sets of criteria to the Canadian Study of Health and Aging (CSHA) data sets are addressed and a procedure for identifying a subgroup presumed "at risk" for developing dementia is presented. Application of an informant's report of changes in cognitive functioning and neuropsychologists' ratings of mild to severe deficits in any of eight cognitive domains results in approximately half of the CIND cases being identified as "at risk." The rationale for the collection of specific information related to CIND in CSHA-2 is provided. A minority of people identified with CIND at CSHA-2 showed only memory impairment, and most demonstrated cognitive loss over the preceding five-year interval. This article provides a conceptual basis for procedures to identify people with cognitive impairment most likely to decline to dementia. PMID- 11892967 TI - Study sampling in the Canadian Study of Health and Aging. AB - The Canadian Study of Health and Aging drew representative samples of people aged 65 or over from the community and institutions across Canada. The sample was designed to provide regional and national prevalence estimates for dementia by age and sex. Thirty-six sampling areas were used in a stratified cluster design with optimal allocation; sampling weights were developed to provide population estimates. The sample included 9,008 people aged 65 or over from the community, and 1,255 from institutions. This report describes the sampling procedures, the methods used to recruit people to the study and participation rates, the characteristics of the resulting sample, and the way in which sample weights should be used. PMID- 11892968 TI - Evaluating screening tests for dementia and cognitive impairment in a heterogeneous population in the presence of verification bias. AB - This article reviews two potentially serious sources of error in the evaluation of screening tests, namely, verification bias and the influence of demographic covariates. It demonstrates how to deal with these problems statistically. Verification bias arises when not all subjects receive a definitive diagnosis following a screening test. If only a small proportion of those who screen negative are sent for diagnosis, the calculated test sensitivity is an overestimate and the calculated specificity an underestimate. The methodology outlined in this article may be extended to psychological and medical screening tests in general. PMID- 11892969 TI - Measurement of the influence of the physical environment on adverse health outcomes: technical report from the Canadian Study of Health and Aging. AB - A paucity of information exists to characterize the relationship between the health status of elderly people and their physical environment. The Canadian Study of Health and Aging (CSHA) is a multicenter study of the distribution of dementia among community-dwelling and institutionalized Canadians aged 65 years and older. The study also provides the opportunity to examine issues such as the physical environment which may be related to the health of elderly people. Six items were used to assess the cleanliness, neatness, and maintenance of the inside and outside of the homes of 8,134 community-dwelling individuals. Data were also obtained to evaluate cognition, physical health, and functional capacity. Five years after the original survey, information pertaining to subsequent institutionalization and/or mortality was obtained. A significant relationship was found between classification of physical environment and the outcomes of institutionalization and mortality. The likelihood of both adverse outcomes was notably higher for individuals living in a "less than ideally maintained environment" compared to an "ideally maintained environment." Limitations of the six items used to assess the physical environment and ways in which to improve the sensitivity of the items, consequently avoiding measurement bias, are discussed. PMID- 11892970 TI - Contribution of self-reported health ratings to predicting frailty, institutionalization, and death over a 5-year period. AB - Cross-sectional data from Phase 1 of the Canadian Study of Health and Aging was used to examine the relationship between two self-report health measures: "How would you say your health is these days?"(HEALTH) and "How much do your health troubles stand in the way of your doing the things you want to do?"(TROUBLE). The contribution of these measures to predictive models for institutionalization and mortality is examined, using linked data from Phases 1 and 2. Their relationship to a proposed frailty measure is also examined. At CSHA-1, a majority of respondents perceived that they were in good health and did not feel that their health problems interfered with their preferred activities. At all frailty levels, a majority of both males and females rated their health as "very good" or "pretty good." As frailty increased, health problems increasingly interfered with normal activities. Logistic regression of the longitudinal data indicated that, despite their correlation, HEALTH and TROUBLE cannot act as proxies for each other. They appear to predict independently; adding one to the other significantly improved prediction of institutionalization and mortality. PMID- 11892971 TI - The Canadian Study of Health and Aging: organizational lessons from a national, multicenter, epidemiologic study. AB - The Canadian Study of Health and Aging was a large, multidisciplinary, national core study--with a number of "add-on" investigations--of the epidemiology of dementia and the health of older people. This structure was a fiscally prudent way to balance between mandated and investigator-initiated inquiry. In hindsight, several important features of the study would be repeated. Future studies might profitably consider a longer funding period for analysis, and a more strategic approach to in-depth, supplementary studies. PMID- 11892972 TI - Data collected in the Canadian Study of Health and Aging. AB - The Canadian Study of Health and Aging collected data focusing on the epidemiology of dementia, using interviews and questionnaires, clinical and neuropsychological examinations, physical measurements and blood collection, and access to public records such as death certificates, from people 65 and over in community (N = 9,008) institutional settings (N = 1,255). The study produced 12 data sets, including community health interviews, clinical and neuropsychological assessments, risk factor questionnaires, and caregiver interviews. This report describes the data collection and processing procedures, summarizes the content of each data set, and outlines the information collected in sufficient detail to permit its suitability for secondary analyses to be scrutinized. PMID- 11892973 TI - Study organization in the Canadian Study of Health and Aging. AB - The Canadian Study of Health and Aging was a complex undertaking that faced management challenges not encountered by smaller-scale projects. The study followed 10,263 elderly people in 18 study centers spanning six time zones; it was administered in two languages, and over 70 investigators were involved. The data collected from each participant were not fixed, but varied according to the results of earlier testing. The data could include a screening interview, a self completed risk factor questionnaire, an interview with a relative, a clinical examination, neuropsychological testing, blood samples, and neuroimaging. This report describes the approach taken to organize the study, to track participants, and to monitor adherence to the study protocol. It also describes the human organizational aspects, including systems for staff training, for communicating among study centers, and for coordinating the publication of results. The discussion proposes some guiding principles for administering multicenter studies. PMID- 11892974 TI - Correlates of nonparticipation in the Canadian Study of Health and Aging. AB - Correlates of nonparticipation in the community interview component of the Canadian Study of Health and Aging and their impact on bias in the results were analyzed. Characteristics of study subjects, their habitats, and encouragement techniques were analyzed to identify correlates of variation in response rates across the 18 study centers. Refusal rates from 14% to 41% varied by age, gender, city size, number of subjects and length of time for enrollment, and method of approach. Cognitively impaired subjects had higher refusal rates which affected prevalence estimates. At one study site, efforts to "convert" subjects who initially refused to participate in the survey were successful with 26% of those who were recontacted. PMID- 11892975 TI - A recommended method for obtaining the age at onset of dementia from the CSHA database. AB - In studies of dementia, the age at onset (AAO) of the disease is often described without indicating how it was obtained. We used the "CAMDEX algorithm," an ad hoc procedure, to compute the AAO of dementia from the CSHA database. An AAO could be calculated for 983 of 1,132 subjects with dementia. A similar procedure (the "clinical algorithm") was used to calculate a second AAO, which was compared to that obtained by the first algorithm. The CAMDEX and clinical algorithms produced mean AAOs of dementia of 80.5 years (SD = 9.2 years, n = 983) and 79.6 years (SD = 9.7 years, n = 829), respectively. The sample correlation coefficient between the CAMDEX and clinical algorithms was .899 while the intraclass correlation coefficient, ICC(2,1), was .898. This method could prove useful for researchers using the CSHA data who need an AAO for those subjects with dementia. PMID- 11892976 TI - An overview of the Canadian Study of Health and Aging. AB - The Canadian Study of Health and Aging is a multicenter, population-based cohort study of dementia with a sample of 10,263 participants aged 65 or over. Field work began in 1991, and a follow-up study was undertaken in 1996-97. The present article describes the origins and objectives of the study, provides an overview of its design, organization, and data collection methods, and offers a brief summary of the main results. PMID- 11892977 TI - Reliable individual change scores on the 3MS in older persons with dementia: results from the Canadian Study of Health and Aging. AB - We examined the degree of interrater agreement on the Modified Mini-Mental State Examination (3MS), administered both in the home and at the clinical examination, to determine the boundaries of reliable individual changes for 257 community dwelling older persons who received a diagnosis of dementia at CSHA-1. Individual score differences were approximately normally distributed (mean of differences 0.2; SD 8.0; 95% confidence interval -16 to 16). The intraclass correlation coefficient was 0.85. Except for the language of testing, there was no relationship between score differences and the determinants investigated (i.e., age, education, type and severity of dementia). This study provides evidence that, in a time frame compatible with no change in cognition, the discrepancy between repeat 3MS scores can be as large as +/- 16. These limits represent the range of variability consistent with no change and should be considered when interpreting individual change scores. PMID- 11892978 TI - Measuring psychological well-being in the Canadian Study of Health and Aging. AB - The Canadian Study of Health and Aging (CHSA) provided an opportunity to examine the positive aspects of aging. CHSA-2 included the 18-item Ryff multidimensional measure of well-being, which taps six core theoretical dimensions of positive psychological functioning. The measure was administered to 4,960 seniors without severe cognitive impairment or dementia at CSHA-2. Intercorrelations across scales were generally low. At the same time, the internal consistency reliability of each of the 6 subscales was not found to be high. Confirmatory factor analyses provide support for a 6-factor model, although some items demonstrate poor factor loadings. The well-being measures in CSHA-2 provide an opportunity to examine broad, descriptive patterns of well-being in Canadian seniors. PMID- 11892979 TI - Imputation of missing dates of death or institutionalization for time-to-event analyses in the Canadian Study of Health and Aging. AB - Data from the Canadian Study of Health and Aging (CSHA) allow investigators to study patterns and predictors of mortality and institutional placement in a well characterized, population-based cohort of elderly Canadians. However, it is impossible to study the timing of these events if the date of occurrence is missing. This technical article describes a procedure for imputing missing dates of death or institutionalization. The first step consists in identifying and correcting dates that are inconsistent with other available dates on which we know the event has or has not occurred. A missing date for an event is then replaced by the middle of a range of plausible dates for its occurrence. This constitutes a valuable addition to the CSHA data since it precludes the loss of information that results from discarding subjects with missing occurrence dates in time-to-event analyses. PMID- 11892980 TI - Estimating antemortem cognitive status of deceased subjects in a longitudinal study of dementia. AB - There was a five-year delay between the two waves of the Canadian Study of Health and Aging during which 2,982 participants died. Their cognitive status before death should be taken into account in estimating the incidence of dementia in the cohort. Information concerning antemortem cognitive status was available from death certificates and from an interview with a close relative of the decedent at the CSHA-2 follow-up. The interview included a direct question on whether the person had been diagnosed with dementia and questions covering cognitive signs and symptoms from which we formed an algorithm to predict probability of dementia. These sources of information were validated using a small sample of study participants who died within five months of undergoing the CSHA clinical examination. Sensitivity of the death certificate and the question regarding diagnosis of dementia was low (33% and 44%), although their specificity was very high. Accordingly, we combined these with the predictive algorithm to form an overall estimate of the probability of antemortem dementia. This raised the sensitivity to 82% (specificity 93%). PMID- 11892981 TI - Molecular biology of stress responses. PMID- 11892982 TI - Overexpression of Hsp25 in K1735 murine melanoma cells enhances susceptibility to natural killer cytotoxicity. AB - In the present study we used a murine melanoma model to investigate the effect of the 25-kDa heat shock protein (Hsp25) on natural killer (NK) cytotoxicity. The melanoma lines K1735-C123 (low metastatic potential) and K1735-M2 (high metastatic potential) were transfected with hsp25 and a control plasmid. Highly purified interleukin (IL)-2-stimulated DX-5+ NK cells showed enhanced lysis of Hsp25-overexpressing K1735-C123 targets in comparison with controls. In contrast, there was no difference in susceptibility to lysis by purified IL-2-stimulated DX 5+ NK cells between Hsp25-overexpressing and control-transfected K1735-M2 targets. Fluorescence-activated cell sorter analysis revealed that Hsp25 is displayed on the cell surface independently of Hsp25 overexpression and metastatic phenotype. Thus, surface localization of Hsp25 does not correlate with the target cell susceptibility to killing. To sum up, a cytoplasmic overexpression of Hsp25 is associated with an increased susceptibility to lysis by DX-5+ NK cells in the low-metastatic murine melanoma model investigated. PMID- 11892983 TI - Molecular characterization of a Bombyx mori protein disulfide isomerase (bPDI). AB - We have isolated a complementary deoxyribonucleic acid clone that encodes the protein disulfide isomerase of Bombyx mori (bPDI). This protein has a putative open reading frame of 494 amino acids and a predicted size of 55.6 kDa. In addition, 2 thioredoxin active sites, each with a CGHC sequence, and an endoplasmic reticulum (ER) retention signal site with a KDEL motif were found at the C-terminal. Both sites are typically found in members of the PDI family of proteins. The expression of bPDI messenger ribonucleic acid (mRNA) was markedly increased during ER stress induced by stimulation with calcium ionophore A23187, tunicamycin, and dithiothreitol, all of which are known to cause an accumulation of unfolded proteins in the ER. We also examined the tissue distribution of bPDI mRNA and found pronounced expression in the fat body of insects. Hormonal regulation studies showed that juvenile hormone, insulin, and a combination of juvenile hormone and transferrin (although not transferrin alone) affected bPDI mRNA expression. A challenge with exogenous bacteria also affected expression, and the effect peaked 16 hours after infection. These results suggest that bPDI is a member of the ER-stress protein group, that it may play an important role in exogenous bacterial infection of the fat body, and that its expression is hormone regulated. PMID- 11892984 TI - Heat shock protein 70 is a potent activator of the human complement system. AB - According to new hypotheses, extracellular heat shock proteins (Hsps) may represent an ancestral danger signal of cellular death or lysis-activating innate immunity. Recent studies demonstrating a dual role for Hsp70 as both a chaperone and cytokine, inducing potent proinflammatory response in human monocytes, provided support for the hypothesis that extracellular Hsp is a messenger of stress. Our previous work focused on the complement-activating ability of human Hsp60. We demonstrated that Hsp60 complexed with specific antibodies induces a strong classical pathway (CP) activation. Here, we show that another chaperone molecule also possesses complement-activating ability. Solid-phase enzyme-linked immunosorbent assay was applied for the experiments. Human Hsp70 activated the CP independently of antibodies. No complement activation was found in the case of human Hsp90. Our data further support the hypothesis that chaperones may messenger stress to other cells. Complement-like molecules and primitive immune cells appeared together early in evolution. A joint action of these arms of innate immunity in response to free chaperones, the most abundant cellular proteins displaying a stress signal, may further strengthen the effectiveness of immune reactions. PMID- 11892986 TI - Acute stress-induced tissue injury in mice: differences between emotional and social stress. AB - Emotional stress affects cellular integrity in many tissues including the heart. Much less is known about the effects of social stress. We studied the effect of emotional (immobilization with or without cold exposure) or social (intermale confrontation) stress in mice. Tissue injury was measured by means of the release of enzyme activities to blood plasma: lactate dehydrogenase (LDH), creatine kinase (CK), aspartate transaminase (AST), and alanine transaminase (ALT). Tape immobilization increased all these activities in the plasma. AST-ALT ratio was also increased in these animals. Electrophoretic analysis of CK isoenzymes showed the appearance of CK-MB. These results indicate that the heart was injured in immobilized mice. Analysis of LDH isoenzymes and measurement of alpha hydroxybutyrate dehydrogenase (HBDH) activity suggests that other tissues, in addition to the heart, contribute to the increase in plasma LDH activity. Restraint in small cylinders increased plasma LDH, CK, AST, and ALT activities, but to lower levels than in tape immobilization. Because the decrease in liver glycogen and the increase in plasma epidermal growth factor (EGF) were also smaller in restraint than in the tape-immobilization model of emotional stress, we conclude that the former is a less intense stressor than the latter. Cold exposure during the restraint period altered the early responses to stress (it enhanced liver glycogen decrease, but abolished the increase in plasma EGF concentration). Cold exposure during restraint enhanced heart injury, as revealed by the greater increase in CK and AST activities. Intermale confrontation progressively decreased liver glycogen content. Plasma EGF concentration increased (to near 100 nM from a resting value of 0.1 nM) until 60 minutes, and decreased thereafter. Confrontation also affected cellular integrity in some tissues, as indicated by the rise in plasma LDH activity. However, in this type of stress, the heart appeared to be specifically protected because there was no increase in plasma CK activity, and both AST and ALT increased, but the AST-ALT ratio remained constant. Habituation to restraint (1 h/d, 4 days) made mice resistant to restraint-induced tissue injury as indicated by the lack of an increase in plasma LDH, CK, AST, or ALT activities. Similar general protection against homotypic stress-induced injury was observed in mice habituated to intermale confrontation. PMID- 11892987 TI - The eukaryote chaperonin CCT is a cold shock protein in Saccharomyces cerevisiae. AB - The eukaryotic Hsp60 cytoplasmic chaperonin CCT (chaperonin containing the T complex polypeptide-1) is essential for growth in budding yeast, and mutations in individual CCT subunits have been shown to affect assembly of tubulin and actin. The present research focused mainly on the expression of the CCT subunits, CCTalpha and CCTbeta, in yeast (Saccharomyces cerevisiae). Previous studies showed that, unlike most other chaperones, CCT in yeast does not undergo induction following heat shock. In this study, messenger ribonucleic acid (mRNA) and protein levels of CCT subunits following exposure to low temperatures, were examined. The Northern blot analysis indicated a 3- to 4-fold increase in mRNA levels of CCTalpha and CCTbeta genes after cold shock at 4 degrees C. Interestingly, Western blot analysis showed that cold shock induces an increase in the CCTalpha protein, which is expressed at 10 degrees C, but not at 4 degrees C. Transfer of 4 degrees C cold-shocked cells to 10 degrees C induced a 5-fold increase in the CCTalpha protein level. By means of fluorescent immunostaining and confocal microscopy, we found CCTalpha to be localized in the cortex and the cell cytoplasm of S. cerevisiae. Localization of CCTalpha was not affected at low temperatures. Co-localization of CCT and filaments of actin and tubulin was not observed by microscopy. The induction pattern of the CCTalpha protein suggests that expression of the chaperonin may be primarily important during the recovery from low temperatures and the transition to growth at higher temperatures, as found for other Hsps during the recovery phase from heat shock. PMID- 11892985 TI - Overexpression of apolipoprotein J in human fibroblasts protects against cytotoxicity and premature senescence induced by ethanol and tert butylhydroperoxide. AB - Human diploid fibroblasts (HDFs) exposed to subcytotoxic stresses under H2O2, tert-butylhydroperoxide (t-BHP), and ethanol (EtOH) undergo stress-induced premature senescence (SIPS) characterized by many biomarkers of HDFs replicative senescence. Among these biomarkers are a growth arrest, an increase in the senescence-associated beta-galactosidase activity, a senescent morphology, an overexpression of p21waf-1 and the subsequent inability to phosphorylate pRb, the presence of the common 4977-bp mitochondrial deletion, and an increase in the steady-state level of several senescence-associated genes such as apolipoprotein J (apo J). Apo J has been described as a survival gene against cytotoxic stress. In order to study whether apo J would be protective against cytotoxicity SIPS and replicative senescence in human fibroblasts, a full-length complementary deoxyribonucleic acid of apo J was transfected into WI-38 HDFs and SV40 transformed WI-38 HDFs. The overexpression of apo J resulted in an increased cell survival after t-BHP and EtOH stresses at cytotoxic concentrations. In addition, when WI-38 HDFs were exposed to 5 subcytotoxic stresses with EtOH or t-BHP, in conditions that were previously shown to induce SIPS, a lower induction of 2 biomarkers of SIPS was observed in HDFs overexpressing apo J. No effect of apo J overexpression was observed on the proliferative life span of HDFs, even if apo J overexpression triggered osteonectin (SPARC) overexpression, which was shown to decrease the mitogenic potential of platelet-derived growth factor but not of other common growth-inducing conditions. Apo J senescence-related overexpression is proposed to have antiapoptotic rather than antiproliferative effects. PMID- 11892988 TI - Xenopus small heat shock proteins, Hsp30C and Hsp30D, maintain heat- and chemically denatured luciferase in a folding-competent state. AB - In this study we characterized the chaperone functions of Xenopus recombinant Hsp30C and Hsp30D by using an in vitro rabbit reticulocyte lysate (RRL) refolding assay system as well as a novel in vivo Xenopus oocyte microinjection assay. Whereas heat- or chemically denaturated luciferase (LUC) did not regain significant enzyme activity when added to RRL or microinjected into Xenopus oocytes, compared with native LUC, denaturation of LUC in the presence of Hsp30C resulted in a reactivation of enzyme activity up to 80-100%. Recombinant Hsp30D, which differs from Hsp30C by 19 amino acids, was not as effective as its isoform in preventing LUC aggregation or maintaining it in a folding-competent state. Removal of the first 17 amino acids from the N-terminal region of Hsp30C had little effect on its ability to maintain LUC in a folding-competent state. However, deletion of the last 25 residues from the C-terminal end dramatically reduced Hsp30C chaperone activity. Coimmunoprecipitation and immunoblot analyses revealed that Hsp30C remained associated with heat-denatured LUC during incubation in reticulocyte lysate and that the C-terminal mutant exhibited reduced affinity for unfolded LUC. Finally, we found that Hsc70 present in RRL interacted only with heat-denatured LUC bound to Hsp30C. These findings demonstrate that Xenopus Hsp30 can maintain denatured target protein in a folding competent state and that the C-terminal end is involved in this function. PMID- 11892989 TI - Differential accumulation of U14 snoRNA and hsc70 mRNA in Chinese hamster cells after exposure to various stress conditions. AB - We have previously characterized the unique organization of the U14 small nucleolar ribonucleic acid (snoRNA) gene in Chinese hamster HA-1 cells. The single copy of the hsc70/U14 gene is the only source for the production of both U14 snoRNA species and hsc70 messenger ribonucleic acid (mRNA) in these cells. Here we report that the accumulations of U14 snoRNA and hsc70 mRNA are different in response to various stress conditions, although both of them are transcribed in a single primary transcript. Heat shock induced an increased accumulation of both U14 snoRNA and hsc70 mRNA. On the other hand, exposure to sodium arsenite or azetidine induced an increased accumulation of hsc70 mRNA, but did not lead to a concomitant increase in the level of U14 snoRNA. Under normal growth conditions, the variations in the levels of U14 snoRNA and hsc70 mRNA, in the different phases of the cell cycle, are correlated. The increased expression of U14 snoRNA and hsc70 mRNA, and the hsc70 protein induced specifically by heat shock suggest that they participate in the repair process of heat-induced damage to macromolecular complexes involved in the synthesis and processing of ribosomal RNA. PMID- 11892990 TI - The effect of hyperthermia on the induction of cell death in brain, testis, and thymus of the adult and developing rat. AB - Stressful stimuli can elicit 2 distinct reactive cellular responses, the heat shock (stress) response and the activation of cell death pathways. Most studies on the effects of hyperthermia on the mammalian nervous system have focused on the heat shock response, characterized by the transient induction of Hsps, which play roles in repair and protective mechanisms. This study examines the effect of hyperthermia on the induction of cell death via apoptosis, assayed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and active caspase 3 cytochemistry, in the adult rat brain, testis, and thymus. Results show that a fever-like increase in temperature triggered apoptosis in dividing cell populations of testis and thymus, but not in mature, postmitotic cells of the adult cerebellum. These differential apoptotic responses did not correlate with whole-tissue levels of Hsp70 induction. We further investigated whether dividing neural cells were more sensitive to heat-induced apoptosis by examining the external granule cell layer of the cerebellum at postnatal day 7 and the neuroepithelial layers of the neocortex and tectum at embryonic day 17. These proliferative neural regions were highly susceptible to hyperthermia induced apoptosis, suggesting that actively dividing cell populations are more prone to cell death induced by hyperthermia than fully differentiated postmitotic neural cells. PMID- 11892991 TI - Hsp90, not Grp94, regulates the intracellular trafficking and stability of nascent ErbB2. AB - The benzoquinone ansamycin geldanamycin (GA) stimulates proteasome-mediated degradation of plasma membrane-associated ErbB2, a receptor tyrosine kinase. Drug sensitivity is mediated by ErbB2's kinase domain and occurs subsequent to the disruption of Hsp90 interaction with this domain. Full-length ErbB2 is efficiently processed via the endoplasmic reticulum (ER) and Golgi network, so that at steady state most of the detectable protein is plasma membrane associated. However, previous studies have also demonstrated the GA sensitivity of newly synthesized ErbB2, normally a minor component of the total cellular pool of the kinase. Drug sensitivity of nascent ErbB2 is distinguished by 2 characteristics--protein instability and inability to traverse the ER. As nascent ErbB2 can associate with both cytoplasmic Hsp90 and its ER luminal homolog Grp 94, also a GA-binding protein, the purpose of this study was to examine the relative contributions of the cytoplasmic and ER luminal domains of ErbB2 to the GA sensitivity of the nascent kinase. By studying the drug sensitivity of ErbB2/DK, a construct lacking ErbB2's cytoplasmic kinase domain, and by examining the activity of a GA derivative that preferentially binds Hsp90, we conclude that both the stability and the maturation of nascent ErbB2 are regulated by its cytoplasmic, Hsp90-interacting domain. PMID- 11892992 TI - Thermal acclimation and stress in the American lobster, Homarus americanus: equivalent temperature shifts elicit unique gene expression patterns for molecular chaperones and polyubiquitin. AB - Using homologous molecular probes, we examined the influence of equivalent temperature shifts on the in vivo expression of genes coding for a constitutive heat shock protein (Hsc70), heat shock proteins (Hsps) (Hsp70 and Hsp90), and polyubiquitin, after acclimation in the American lobster, Homarus americanus. We acclimated sibling, intermolt, juvenile male lobsters to thermal regimes experienced during overwintering conditions (0.4 +/- 0.3 degrees C), and to ambient Pacific Ocean temperatures (13.6 +/- 1.2 degrees C), for 4-5 weeks. Both groups were subjected to an acute thermal stress of 13.0 degrees C, a temperature shift previously found to elicit a robust heat shock response in ambient acclimated lobsters. Animals were examined after several durations of acute heat shock (0.25-2 hours) and after several recovery periods (2-48 hours) at the previous acclimation temperature, following a 2-hour heat shock. Significant inductions in Hsp70, Hsp90, and polyubiquitin messenger RNA (mRNA) levels were found for the ambient-acclimated group. Alternatively, for the cold-acclimated group, an acute thermal stress over an equivalent interval resulted in no induction in mRNA levels for any of the genes examined. For the ambient acclimated group, measurements of polyubiquitin mRNA levels showed that hepatopancreas, a digestive tissue, incurred greater irreversible protein damage relative to the abdominal muscle, a tissue possessing superior stability over the thermal intervals tested. PMID- 11892993 TI - The neurobiology of dopamine signaling. AB - The biochemistry of synaptic transmission, especially the neurobiology of dopamine signaling, is discussed. PMID- 11892995 TI - The relationship between the reference value of erythrocyte sedimentation rate and geographical factors. AB - In order to provide a scientific basis for a unified standard of the reference value of erythrocyte sedimentation rate (ESR) in China, the reference value of healthy people's ESR has been collected. The relationship between the reference value of ESR and geographical factors is examined in this paper. Altitude is the most important factor affecting the reference value of ESR, which decreases with increasing altitude; the relationship is quite significant. The method of stepwise regression analysis was used to deduce two regression equations: Y1 = 12.08 - 0.00222X1 + 0.00114X5 +/- 2.95, Y2 = 18.81 - 0.00323X1 + 0.239X4 +/- 4.70. If the geographical factor value of a particular area in China is known, the reference value of ESR of Chinese people can be calculated by means of the regression equations. Furthermore, the dependence on geographical factors in China can be classified as six districts: Qingzang, Southwest, Northwest, Southeast, North and Northeast. PMID- 11892994 TI - Liver lipid composition and antioxidant enzyme activities of spontaneously hypertensive rats after ingestion of dietary fats (fish, olive and high-oleic sunflower oils). AB - Hypertension is associated with greater than normal lipoperoxidation and an imbalance in antioxidant status, suggesting that oxidative stress is important in the pathogenesis of this disease. Although many studies have examined the effect of antioxidants in the diet on hypertension and other disorders, less attention has been given to the evaluation of the role of specific dietary lipids in modulating endogenous antioxidant enzyme status. Previously, we have described that liver antioxidant enzyme activities may be modulated by consumption of different oils in normotensive rats. The purpose of the present study was to examine the effects of feeding different lipidic diets (olive oil, OO, high-oleic acid sunflower oil, HOSO, and fish oil, FO) on liver antioxidant enzyme activities of spontaneously hypertensive rats (SHR). Plasma and liver lipid composition was also studied. Total triacylglycerol concentration increases in plasma and liver of animals fed on the HOSO and OO diets and decreases in those fed on the FO diet, relative to rats fed the control diet. The animals fed on the oil-enriched diet show similar hepatic cholesterol and phospholipid contents, which are higher than the control group. Consumption of the FO diet results in a decrease in the total cholesterol and phospholipid concentration in plasma, compared with the high-oleic-acid diets. In liver, the FO group show higher levels of polyunsaturated fatty acids (PUFA) of the (n - 3) series, in relation to the animals fed on the diets enriched in oleic acid. Livers of FO-fed rats, compared with those of OO- and HOSO-fed rats showed: (i) significantly higher activities of catalase, glutathione peroxidase and Cu/Zn superoxide dismutase; (ii) no differences in the NADPH-cytochrome c reductase activity. The HOSO diet had a similar effect on liver antioxidant enzyme activities as the OO diet. In conclusion, it appears that changes in the liver fatty acid composition due mainly to n - 3 lipids may enhance the efficiency of the antioxidant defence system and may yield a benefit in the hypertension status. The two monounsaturated fatty acids oils studied (OO and HOSO), with the same high content of oleic acid, but different content of natural antioxidants, had similar effects on the antioxidant enzyme activities studied. PMID- 11892996 TI - Fusion and infection of influenza and Sendai viruses as modulated by dextran sulfate: a comparative study. AB - We have directly compared the effect of two types of dextran sulfate with distinct molecular weights (500 kDa and 5 kDa) on the fusion activity and infectivity of both Sendai and influenza viruses, two lipid-enveloped viruses that differ in their routes of entry into target cells. To correlate membrane merging and infectivity MDCK cells were used as targets for the viruses in both approaches. In either case pronounced inhibition of virus-cell interactions by dextran sulfate was only observed at low pH, even though Sendai virus fuses maximally at pH 7.4. Although membrane merging could not be fully abolished, the inhibitory effect was always greater when the higher molecular weight dextran sulfate was used. The presence of this residual fusion activity, that could not be reduced even with high concentrations of agent, suggests that a limited number of binding sites for dextran sulfate may exist on the viral envelopes. The compounds also inhibited fusion of bound virions, and all results could be reproduced using erythrocyte ghosts as target membranes in the fusion assay, instead of MDCK cells. In agreement with these observations only the infectivity of influenza virus (which requires a low pH-dependent step to enter target cells) was affected by dextran sulfate, again the higher molecular weight compound showing a more pronounced inhibitory effect. PMID- 11892997 TI - Ionic charge, hydrophobicity and tryptophan fluorescence of the folate binding protein isolated from cow's milk. AB - A high-affinity folate binding protein was isolated and purified from cow's milk by a combination of cation exchange chromatography and methotrexate affinity chromatography. Chromatofocusing studies revealed that the protein possessed isoelectric points in the pH-interval 8-7. Polymers of the protein prevailing at pH values close to the isoelectric points seemed to be more hydrophobic than monomers present at pH 5.0 as evidenced by hydrophobic interaction chromatography and turbidity (absorbance at 340 nm) in aqueous buffer solutions (pH 5-8). Ligand binding seemed to induce a conformation change that decreased the hydrophobicity of the protein. In addition, Ligand binding quenched the tryptophan fluorescence of folate binding protein suggesting that tryptophan is present at the binding site and/or ligand binding induces a conformation change that affects tryptophan environment in the protein. There was a noticeable discordance between the ability of individual folate analogues to compete with folate for binding and the quenching effect. PMID- 11892998 TI - Mouse beta-mannosidase: cDNA cloning, expression, and chromosomal localization. AB - Beta-mannosidase is an exoglycosidase involved in the degradation of N-linked oligosaccharides moieties of glycoproteins. Lack of beta-mannosidase activity leads to the lysosomal disorder beta-mannosidosis (MIM 248510). We have isolated and sequenced the gene encoding the mouse beta-mannosidase. Comparison of the deduced amino acid sequence of mouse, human, bovine, and goat beta-mannosidase showed 64%, identity, reflecting a high degree of evolutionary conservation. Analysis of a multiple tissue northern blotting revealed a major transcript of about 3.7 kb in all tissues examined. The northern analysis also demonstrates that there is differential tissue mRNA expression. The mouse beta-mannosidase gene (Bmn) was mapped to the distal end of Chromosome (Chr) 3, in a region that is homologous with a segment of human Chr 4 containing the orthologous human gene. PMID- 11892999 TI - Interleukin-6 enhances transforming growth factor-alpha mRNA expression in macrophage-like human monocytoid (U-937-1) cells. AB - We have previously reported that the human monocytoid cell line U-937-1 constitutively expresses transforming growth factor-alpha (TGF-alpha) and that the steady-state levels of TGF-alpha mRNA as well as TGF-alpha protein release increase when U-937-1 cells are differentiated towards monocytes/macrophages. Interleukin-6 (IL-6), which has been shown to have growth-stimulatory effects on a number of cell types, has recently been shown to enhance TGF-alpha expression in keratinocytes. In the present study we investigated whether TGF-alpha expression in macrophage-like cells could be regulated by IL-6 using U-937-1 cells as a model system of monocyte/macrophage differentiation. U-937-1 cells were differentiated with retinoic acid (RA), vitamin D3 (Vit-D3) or phorbol-12 myristate-13-acetate (PMA) for 4 days and were then treated with human recombinant IL-6 (1000 IU/ml) for up to 24 hr. Northern blot analysis revealed that cells differentiated with PMA, inducing the phenotype of a secretory macrophage, markedly increased their TGF-alpha mRNA levels (2.7-fold) when treated with IL-6; the response was maximal at 6 hr and remained high at 12 hr. The expression of the TGF-alpha gene was accompanied by release of TGF-alpha protein into the cell culture medium, irrespective of differentiating agent, as demonstrated by enzyme-linked immunosorbent assay (ELISA), as well as by surface expression of pro-TGF-alpha as determined by indirect immunofluorescent cytometry. However, the superinduction of the TGF-alpha gene by IL-6 in cells differentiated with PMA was not accompanied by any increase in TGF-alpha protein release or pro-TGF-alpha surface expression. We conclude that since IL-6 causes increased steady-state levels of TGF-alpha mRNA in macrophage-like cells, it may prime these cells for production of this growth factor. Furthermore, we have shown that the IL-6 receptor complex is functional in U-937-1 cells induced to differentiate towards a secretory macrophage by treatment with PMA. PMID- 11893000 TI - Real-time measurement of intracellular reactive oxygen species using Mito tracker orange (CMH2TMRos). AB - We have investigated a novel method to monitor real changes of intracellular ROS by the use of CMH2TMRos (a reduced form of MitoTracker orange) in Swiss 3T3 fibroblasts. Arachidonic acid induced a rapid increase of CMTMRos fluorescence with a maximal elevation at 120-150 sec, which was determined by scanning every 10 sec with a confocal microscope. The fluorescence increase by arachidonic acid was completely inhibited by 2-MPG but not by catalase, indicating a major contribution of superoxide to the oxidation of CMH2TMRos. Incubation with glucose oxidase, exogenous H2O2, KO2 and lysophosphatidic acid also increased the CMTMRos fluorescence, which was blocked by 2-MPG. These results suggested that CMH2TMRos is a useful fluorophore for real-time monitoring of intracellular ROS and also indicated that CMH2TMRos detects primarily superoxide in cells even though the fluorophore can be oxidized by both superoxide and H2O2. PMID- 11893001 TI - Antimalarial drugs exacerbate rat liver microsomal lipid peroxidation in the presence of oxidants. AB - The study was undertaken to evaluate the effect of prior treatment of rats with the antimalarial drugs amodiaquine (AQ) mefloquine (MQ) and halofantrine (HF) on rat liver microsomal lipid peroxidation in the presence of 1 mM FeSO4, 1 mM ascorbate and 0.2 mM H2O2 (oxidants). Ingestion of alpha-tocopherol, a radical chain-breaking antioxidant was also included to assess the role of antioxidants in the drug treatment. In the presence of oxidants AQ, MQ and HF elicited 288%, 175% and 225% increases in malondialdehyde (MDA) formation while the drugs induced 125%, 63% and 31% increases in the absence of oxidants respectively. Similarly, AQ, MQ and HF induced lipid hydroperoxide formation by 380%, 256%, 360% respectively in the presence of oxidants and 172%, 136% and 92% in the absence of exogenously added oxidants respectively. a-tocopherol reduced AQ, MQ and HF-induced MDA formation by 40%, 55% and 52% respectively and lipid hydroperoxide formation by 53%, 59% and 54% respectively. Similarly, alpha tocopherol attenuated the AQ, MQ and HF-induced MDA formation by 49%, 51% and 51% in the presence of oxidants and lipid hydroperoxide formation by 61%, 62% and 47% respectively. The results indicate that rat liver microsomal lipid peroxidation could be enhanced by antimalarial drugs in the presence of reactive oxygen species and this effect could be ameliorated by treatment with antioxidants. PMID- 11893002 TI - Expression of winged bean basic agglutinin in Spodoptera frugiperda insect cell expression system. AB - In this paper we report the successful expression of the winged bean basic agglutinin (WBA I) in insect cells infected with a recombinant baculovirus carrying the WBA I gene and its characterization in terms of its carbohydrate binding properties. The expressed protein appears to have a lower molecular weight than the native counterpart which is consistent with the lack of glycosylation of the former. Moreover, the expressed protein maintains its dimeric nature. Hence, a role for glycosylation in modulation of dimerization of WBA I is ruled out unlike Erythrina corallodendron (EcorL). Despite this the protein is active, with its sugar specificity unaltered. PMID- 11893003 TI - Stress-induced changes in ubiquinone concentration and alternative oxidase in plant mitochondria. AB - We have investigated the influence of stress conditions such as incubation at 4 degrees C and incubation in hyperoxygen atmosphere, on plant tissues. The ubiquinone (Q) content and respiratory activity of purified mitochondria was studied. The rate of respiration of mitochondria isolated from cold-treated green bell peppers (Capsicum annuum L) exceeds that of controls, but this is not so for mitochondria isolated from cold-treated cauliflower (Brassica oleracea L). Treatment with high oxygen does not alter respiration rates of cauliflower mitochondria. Analysis of kinetic data relating oxygen uptake with Q reduction in mitochondria isolated from tissue incubated at 4 degrees C (bell peppers and cauliflowers) and at high oxygen levels (cauliflowers) reveals an increase in the total amount of Q and in the percentage of inoxidizable QH2. The effects are not invariably accompanied by an induction of the alternative oxidase (AOX). In those mitochondria where the AOX is induced (cold-treated bell pepper and cauliflower treated with high oxygen) superoxide production is lower than in the control. The role of reduced Q accumulation and AOX induction in the defense against oxidative damage is discussed. PMID- 11893004 TI - Leukodystrophy and CSF purine abnormalities associated with isolated 3 methylcrotonyl-CoA carboxylase deficiency. AB - We report the first case of isolated biotin resistant 3-methylcrotonyl-CoA carboxylase (MCC) deficiency in Argentina. The diagnosis was established at 14 months of age by urinary organic-acid analysis and confirmed by enzyme assay in fibroblasts. The patient suffered from severe psychomotor retardation, hypotonia, areflexia, and failure to thrive, and died unexpectedly at 3 years 4 months of life. Brain MRI at 14 months showed signals of the white matter on cerebral T2 weighted, which were indicative of confluent and multiple foci of leukodystrophy, a pattern not previously described in this entity. In addition, high levels of oxypurines were detected in cerebrospinal fluid. This might be related to energetic consequences of the enzyme deficiency in the brain. This case extends the phenotype of isolated MCC deficiency in infancy and suggests this entity should be considered to be one of the possible causes of "metabolic leukodystrophies." PMID- 11893005 TI - Quantification of the electroencephalographic theta/alpha ratio for the assessment of portal-systemic encephalopathy following implantation of transjugular intrahepatic portosystemic stent shunt (TIPSS). AB - The aim of the study was the quantification of metabolically caused electroencephalographic changes of portal-systemic encephalopathy, a prototype of hepatic encephalopathy. We examined 12 patients with liver cirrhosis before and after implantation of a transjugular intrahepatic portosystemic stent shunt (TIPSS) by means of quantitative digital electroencephalography (EEG). One month after TIPSS implantation, all patients showed an increase in the power of the theta frequency band as well as a decrease in the power of the alpha frequency band. To reduce the error variance, we formed the quotient of the relative power of the theta and alpha frequency band. Theta/alpha quotient values over 0.7 indicate a general change of the EEG with a sensitivity of 93% and a specificity of 87%. The results we have to hand indicate a correlation between the albumin concentration and the theta/alpha quotient 1 and 3 months after TIPSS. No significant correlation was revealed with regard to the Child-Pugh score or the liver function parameters cholinesterase, bilirubin, and prothrombin time. Neither the arterial ammonia concentration nor the performance in the psychometric test showed significance in relation to the theta/alpha quotient. Substances with a high albumin bond and potential neurotoxicity may--in the case of lower albumin levels--be absorbed with increased frequency in the CNS and may be responsible for the observed EEG change. PMID- 11893006 TI - Correlations between cerebral glucose metabolism and neuropsychological test performance in nonalcoholic cirrhotics. AB - Many cirrhotics have abnormal neuropsychological test scores. To define the anatomical-physiological basis for encephalopathy in nonalcoholic cirrhotics, we performed resting-state fluorodeoxyglucose positron emission tomographic scans and administered a neuropsychological test battery to 18 patients and 10 controls. Statistical parametric mapping correlated changes in regional glucose metabolism with performance on the individual tests and a composite battery score. In patients without overt encephalopathy, poor performance correlated with reductions in metabolism in the anterior cingulate. In all patients, poor performance on the battery was positively correlated (p < 0.001) with glucose metabolism in bifrontal and biparietal regions of the cerebral cortex and negatively correlated with metabolism in hippocampal, lingual, and fusiform gyri and the posterior putamen. Similar patterns of abnormal metabolism were found when comparing the patients to 10 controls. Metabolic abnormalities in the anterior attention system and association cortices mediating executive and integrative function form the pathophysiological basis for mild hepatic encephalopathy. PMID- 11893007 TI - Increased expression of "peripheral-type" benzodiazepine receptors in human temporal lobe epilepsy: implications for PET imaging of hippocampal sclerosis. AB - Increased binding sites for "peripheral-type" benzodiazepine receptor (PTBR) ligands have been described in a wide range of neurological disorders including both human and experimental epilepsy. This study was undertaken to assess PTBR expression in relation to the presence of hippocampal sclerosis in human temporal lobe epilepsy (TLE). For this purpose, hippocampal CA1 subfields were dissected from surgical samples from patients with therapy-refractive TLE with (n = 5) or without (n = 2) hippocampal sclerosis and from age-matched nonepileptic postmortem controls (n = 5). PTBR expression was assessed by immunohistochemistry and reverse-transcription polymerase chain reaction. Receptor sites were evaluated using an in vitro binding assay and the selective PTBR ligand [3H]PK11195. Epileptic patients with hippocampal sclerosis showed increases in PTBR binding sites, immunoreactivity, and mRNA expression compared to both nonsclerotic TLE patients and postmortem nonepileptic controls. Induction of PTBR expression and binding sites were directly correlated with the presence of hippocampal sclerosis and the accompanying reactive gliosis. PMID- 11893008 TI - Respiratory chain complex-I defect mimicking myasthenia. AB - In a 67-year-old woman with ptosis, double vision, dysphagia, ambiguous Tensilon tests, normal acetylcholine-receptor antibodies, normal thymus, and repeatedly abnormal responses to low-frequency repetitive stimulation, ocular myasthenia was suspected. Pyridostigmin was ineffective, but corticosteroids improved the abnormalities. Despite this therapy, lower-limb weakness developed. Reevaluation disclosed abnormal increase of serum lactate during slight exercise, myogenic electromyography, ragged-red fibers, reduced oxidative enzyme staining and abnormally shaped and structured mitochondria on muscle biopsy, and a respiratory chain complex-I defect on biochemical investigation of the muscle homogenate. Respiratory chain disorder due to complex-I defect with abnormal decremental response to low-frequency repetitive stimulation was diagnosed. It is concluded that respiratory chain disorders due to a complex-I defect may mimic ocular myasthenia clinically, electrophysiologically, and even therapeutically. PMID- 11893009 TI - Cessation of low-dose gonadotropin releasing hormone agonist therapy followed by high-dose gonadotropin stimulation yields a favorable ovarian response in poor responders. AB - PURPOSE: This study is a prospective nonrandomized study to determine the effect of a new protocol of controlled ovarian hyperstimulation (COH) using low doses and a half-period of gonadotropin releasing hormone agonist (GnRHa) followed by high doses of gonadotropin in patients who were supposed to be poor responders to standard long protocols of GnRHa administration. METHODS: From Dec 1996 to Nov 1998, 50 patients who were classified as "poor responders" were scheduled for 52 cycles of a modified controlled ovarian hyperstimulation protocol. They were categorized into 3 groups: a group of poor responders to COH in the previous IVF or IUI cycles, a group with elevated Day 3 FSH levels, and a group over the age of 40 years. All patients received GnRH agonist from the midluteal phase of the previous cycle to the onset of menstruation in the next cycle. Then high doses of gonadotropins (HMG/FSH) were given. The patients then had standard courses of in vitro fertilization and embryo transfer (IVF-ET) or transfallopian embryo transfer (TET). RESULTS: Six of the 52 cycles of the modified protocols were cancelled because of poor ovarian response. One premature ovulation was noted before ovum retrieval was performed. In the other 45 cycles, an average of 6.3 mature oocytes were retrieved. The total pregnancy rate and implantation rate were 20.5 and 11.5%, respectively. CONCLUSIONS: The low dose and half duration of GnRHa therapy lessened the suppression of the response of the ovaries to COH compared with the regular long protocol of GnRHa down regulation therapy. This resulted in a low cancellation rate (11.8%), a favorable embryo implantation rate (11.5%), and an acceptable clinical pregnancy rate (20.5%). PMID- 11893010 TI - The effect of ease of transfer and type of catheter used on pregnancy and implantation rates in an IVF program. AB - PURPOSE: To test the effects of type of embryo transfer catheter, transfer difficulty, and observations after the transfer procedure on pregnancy and implantation rates in an IVF programme. METHODS: Patients were prepared for IVF using standard protocols. Embryo transfer was performed using either Edwards Wallace or TDT catheter. The difficulty of transfer was graded by a clinician and biologist. Blood observed inside the catheter after the transfer procedure was scored as endometrial damage. Pregnancy and implantation rates were scored. RESULTS: Type of embryo transfer catheter and the observation of blood did not significantly affect pregnancy and implantation rates when transfer was performed by a single operator. CONCLUSIONS: In the hands of experienced, skilled operators, neither choice of transfer catheter and difficulty of transfer nor observations of blood on the transfer catheter caused any significant reduction in outcome to the patient. PMID- 11893011 TI - Vascular endothelial growth factor concentrations in follicular fluid obtained from IVF-ET patients: a comparison of hMG, clomiphene citrate, and natural cycle. AB - PURPOSE: Evaluation of vascular endothelial growth factor (VEGF) concentrations of follicular fluid (FF) in accordance with an ovarian stimulation protocol and ovarian response. METHODS: The subjects of this study were 38 patients undergoing IVF-ET in our hospital. They were divided into three groups according to ovarian stimulation protocols; Group 1: hMG cycles (n = 19), Group 2: clomiphene citrate cycles (n = 10), Group 3: natural cycles (n = 9). They reclassified into three groups according to the number of oocytes harvested. VEGF concentration was measured in FF, employing ELISAs. RESULTS: Group 1 shows lower VEGF concentrations in FF than Group 2 or Group 3. Excluding high responders from Group 1, no difference was found among these three groups. As for the reclassified groups, group of highest number of oocytes havested showed lowest VEGF concentrations in FF. CONCLUSIONS: VEGF concentrations in FF might negatively correlate with the number of follicles, irrespective of the ovulation induction protocol. PMID- 11893012 TI - Expression and function of 3beta hydroxisteroid dehydrogenase (3beta HSD) type II and corticosteroid binding globulin (CBG) in granulosa cells from ovaries of women with and without endometriosis. AB - PURPOSE: To investigate the secretion of progesterone (P4) and corticosteroid binding globulin (CBG) by granulosa luteal cells (GC) as well as the mRNA levels of CBG and 3beta hydroxisteroid dehydrogenase (3beta HSD), in women with and without endometriosis in vivo and in vitro. METHODS: Prospective study in a private, university-affiliated assisted reproduction unit, including women with severe endometriosis (n = 14) or without the disease (n = 20) undergoing in vitro fertilization/intracytoplasmic sperm injection and embryo transfer. GC were obtained from each follicle aspirated, pooled for each patient, and follicular and blood contaminating leukocytes depleted through immunomagnetic purification. Secreted P4 and CBG, and mRNA for both 3beta HSD and CBG were determined in vivo and in vitro using RIA and reverse transcription followed by competitive polymerase chain reaction (cRT-PCR). RESULTS: The pattern of expression of 3beta HSD and CBG mRNAs in vivo and in vitro was similar in both groups. Also, GC from patients with endometriosis produced equal amounts of P4 and CBG than controls without the disease, either in freshly isolated cells or in 24-h cultures. CONCLUSIONS: The GC function in terms of 3beta HSD and CBG mRNA expression and P4/CBG secretion does not seem to be altered in patients with endometriosis in comparison with those without this condition. PMID- 11893013 TI - Improvement of development of vitrified two-cell mouse embryos by vero cell coculture. AB - PURPOSE: The purpose of this study was to determine the developmental potential of two-cell mouse embryos resulting from vitrification could increased using monolayer of Vero cells. METHODS: Two-cell mouse embryos were divided into vitrified and nonvitrified groups. Embryos in the vitrified group were frozen with a combination of 10% ethylene glycol, 30% ficoll, and 0.5% M sucrose (EFS10) as cryoprotectants, and thawed rapidly with 0.5 M sucrose. The survived embryos were cultured either with Vero cells monolayer or in T6 medium. Accordingly the embryos of the nonvitrified group were also cultured. The rates of the development in all the groups were daily determined and statistically compared. At the end of the cultivation period, several expanded blastocysts from each group were stained with ethidium bromide and the mean number of the blastomers were counted and statistically compared. RESULTS: After 4 days of culture, the developmental potential of vitrified-thawed embryos were significantly reduced in Vero cell-free medium, and the mean cell number of embryos reaching the expanded blastocyst stage were also lower than that of nonvitrified embryos. With exception of last day of culture, Vero cell coculture, resulted in a significant increase in the rate of development of vitrified-thawed embryos as well as improved the mean cell number of expanded blastocysts. On the other hand, the mean cell number of expanded blastocysts of nonvitrified group was significantly improved in coculture group. However, the rate of embryo development except for the first day of culture was similar to that of medium alone. CONCLUSIONS: The developmental potential of vitrified-thawed embryos appears to be retarded in conventional medium and Vero cell monolayer is capable to eliminate the postthaw deleterious effect of vitrification during the first 3 days of cultivation, but not for a longer period. PMID- 11893014 TI - A prospective novel method of determining ovarian size during in vitro fertilization cycles. AB - PURPOSE: Recently ovarian volume has been touted as a means to evaluate ovarian reserve in assisted reproductive technology cycles. In this study, a novel method of determining ovarian size was evaluated and compared to the standard three dimensional ovarian volume measurement during in vitro fertilization (IVF). METHODS: This prospective observational study consisted of 60 consecutive patients undergoing baseline transvaginal ultrasonography for IVF from July to August, 1999. The main outcome measures were mean ovarian size and mean ovarian volume. RESULTS: The patients' ages ranged from 23 to 43 years with a mean age of 33.86 +/- 4.5 years. The mean ovarian size was 2.19 +/- 0.4 cm (range 1.40-3.40). The mean ovarian volume was 5.02 +/- 2.7 cm3 (range 1.71-16.5 cm3). By linear regression there was a 90% correlation between the two methods of ovarian measurement (r = 0.90, p < 0.01). CONCLUSIONS: These results demonstrated a strong correlation between these two methods of determining ovarian size. Mean ovarian diameter measured in the largest sagittal plane is a good estimation of ovarian volume and may be used to quickly assess ovarian status prior to undergoing IVF. PMID- 11893015 TI - Tubo-ovarian abscess presenting as pneumoperitoneum. AB - BACKGROUND: Tubo-ovarian abscess (TOA), a serious complication of pelvic inflammatory disease, often require the antibiotic administration, surgical resection or the transvaginal aspiration. Pneumoperitoneum is often associated with the bowel perforation. We reported one case with TOA and pneumoperitoneum that have been mistaken for a perforated bowel with concomitant adnexal mass. CASE: A 30-year-old diabetic Chinese woman was transferred for diffused abdominal pain, mild fever, nausea, and low-grade fever for 5 days. The sonography revealed a 5-cm adnexal mass. The chest X-rays revealed the pneumoperitoneum. Under the impression of bowel perforation and concomitant adnexal cyst, the emergent laparotomy was performed and the TOA was resected. No evidence of gastrointestinal perforation was present. Culture studies showed Escherichia coli without other bacteria flora. The postoperative course was uneventful. CONCLUSION: We concluded that, beside the bowel perforation, TOA should be considered when a diabetic woman presents with pneumoperitoneum and adnexal mass. PMID- 11893016 TI - First report on pregnancies happening in the tube, inside the uterus, and in the cervix. PMID- 11893017 TI - Improved development of very-poor-quality human preembryos by coculture with human fallopian ampullary cells. AB - PURPOSE: To determine whether a confluent culture of fallopian ampullary epithelial cells, taken from women at the end of their reproductive life, is capable of rescuing very-poor-quality preembryos from cleavage arrest and/or degeneration. METHODS: Human preembryos. rejected for transfer or freezing because of very poor quality, and arrested within 24 h of cleavage, were cultured for 5 days in medium alone or over a confluent culture of fallopian ampullary epithelia] cells. Morphological criteria were utilized to assess preembryo degeneration and stage of development. RESULTS: The described coculture rescued preembryos from degeneration, enhancing development to the blastocyst stage 2.2 fold, compared with cultures in medium alone. Furthermore, fully expanded and hatching blastocysts were observed only under coculture conditions. CONCLUSIONS: Very-poor-quality human preembryos may be rescued from degeneration, and their growth and development dramatically improved, when cocultured with a confluent culture of fallopian ampullary epithelial cells. PMID- 11893018 TI - Motivation to join and benefits from participation in parent mutual aid organizations. AB - Findings from the Parent Mutual Aid Organizations (PMAOs) in the Child Welfare Demonstration Project are presented with particular emphasis on parents' motivation to join PMAOs and the benefits from their involvement. Members' perceptions from qualitative investigations are compared with a controlled outcome study. PMAO members show less loneliness, more enthusiasm about their lives, and improved self-esteem and confidence. PMAOs result in substantially less reliance on child protection professionals, fewer child placements, and cost savings for the host agency. PMID- 11893019 TI - Questioning strategies in interviews with children who may have been sexually abused. AB - This article examines the number and types of questions employed in clinical and computer-assisted interviews with children referred for sexual abuse evaluation. This research was part of a larger study to assess the efficacy of a computer assisted protocol in the evaluation of child sexual abuse allegations. Interviews of 47 girls and 29 boys, ages 5 to 10 years, referred to a multidisciplinary clinic for sexual abuse assessment, were analyzed. A coding system was developed from interview transcripts. Nine types of questions were defined. Results indicate that during the initial interview children were asked an average of 195 questions (SD = 92) and that more than 85% of interviewer queries were open ended. The majority of children who disclosed did so in response to focused questions. Findings suggest that many children are able to describe sexual abuse with careful questioning that includes nonleading but focused inquiry. Implications for practice and interviewing guidelines are discussed. PMID- 11893020 TI - Beyond termination: length of stay and predictors of permanency for legally free children. AB - Despite growing awareness of the complex needs of legally free children in child welfare care, relatively little empirical investigation has been done of these children and their experiences. This article reports the results of a study of length of stay and predictors of permanency in a sample of 1,366 legally free children in state custody in Washington state. A Cox proportional-hazards event history model was developed to explore the effects of gender, race, and ethnicity, and age at initial placement, on the likelihood of achieving legalized adoption or guardianship. Results indicate that older children, boys, and African American children were all significantly less likely to achieve a permanent outcome than were Caucasian children. Hispanic children, in the other hand, were significantly more likely to achieve a permanent outcome. The implications of these findings for permanency planning practice and policy development are discussed. PMID- 11893021 TI - Development and characterization of a novel, graded model of clip compressive spinal cord injury in the mouse: Part 1. Clip design, behavioral outcomes, and histopathology. AB - In order to take advantage of various genetically manipulated mice available to study the pathophysiology of spinal cord injury (SCI), we adapted an extradural clip compression injury model to the mouse (FEJOTA mouse clip). The dimensions of the modified aneurysm clip blades were customized for application to the mouse spinal cord. Three clips with different springs were made to produce differing magnitudes of closing force (3, 8, and 24 g). The clips were calibrated regularly to ensure that the closing force remained constant. The surgical procedure involved a laminectomy at T3 and T4, followed by extradural application of the clip at this level for 1 min to produce SCI. Three injury severities (3, 8, and 24 g), sham (passage of dissector extradurally at T3-4), and transection control groups were examined (n = 12/group). Quantitative behavioural assessments using the Basso, Beattie, and Bresnahan (BBB; H > 46; df = 4; p < 0.001; Kruskal-Wallis one-way ANOVA) and inclined plane (IP; F = 123; df = 4; p < 0.0001; two-way repeated measures ANOVA) tests showed a significant graded increase in neurological deficits with increasing severity of injury. By day 14, the motor recovery of the mice plateaued. Qualitative examination of the injury site morphology indicated that microcystic cavitation, degenerating axons, and robust astrogliosis were characteristic of the murine response to clip compressive SCI. Morphometric analyses of H&E/Luxol Fast Blue stained sections at every 50 microm from the injury epicenter indicated that with greater injury severity there was a progressive decrease in residual tissue (F = 220, df = 3; p < 0.0001; two-way ANOVA). In addition, statistically significant differences were found in the amount of residual tissue at the injury epicenter between all of the injury severities (p < 0.05, SNK test). This novel, graded compressive model of SCI will facilitate future studies of the pathological mechanisms of SCI using transgenic and knockout murine systems. PMID- 11893022 TI - Development and characterization of a novel, graded model of clip compressive spinal cord injury in the mouse: Part 2. Quantitative neuroanatomical assessment and analysis of the relationships between axonal tracts, residual tissue, and locomotor recovery. AB - A detailed examination of the histopathological features of the clip compression injury in mice was performed to understand the relationships between neurological function and existing pathology of the spinal cord. Adult, female CD1 mice underwent three grades of extradural clip compression injury (3-g, 8-g, and 24-g FEJOTA mouse clips), transection, and sham injury at T3-4. Quantitative behavioural assessments were performed for 4 weeks following SCI. After 4 weeks, Fluoro-Gold was introduced caudal to the SCI site, at T9, and was retrogradely transported for 5 days to the origin of spared axons through the injury site. Counts of retrogradely labeled neurons in the brain-stem, midbrain, and sensory motor cortex indicated that the number of intact descending axons that traversed the lesion decreased with increasing injury severity (F > 28; df = 4; p < 0.0001; one-way ANOVA). Independent linear correlation analyses were performed between indices of neurological recovery (BBB and IP test), counts of retrogradely labeled neurons and morphometric assessments of normal residual tissue at the injury epicenter. The BBB test correlated strongly with the amount of residual tissue at the injury epicenter (R = 0.945, df = 28, p < 0.0001). Counts of neurons retrogradely labeled with Fluoro-Gold were also strongly correlated with the BBB scores. The extrapyramidal (raphespinal, reticulospinal, vestibulospinal, and rubrospinal) tracts had Pearson correlation coefficients (R) of 0.814, 0.812, 0.813, and 0.747, respectively (df = 28, p < 0.0001). The pyramidal (corticospinal) tract had a correlation of R = 0.747, df = 28, p < 0.0001 with the BBB scores. The IP scores also correlated strongly with the persistence of extrapyramidal (raphespinal, reticulospinal, vestibulospinal and rubrospinal) tracts with correlation coefficients of 0.801, 0.782, 0.790, and 0.836, respectively (df = 28, p < 0.0001). These data indicate that the counts of retrogradely labeled neurons at the origin of distinct descending motor pathways are predictors of the variance of the functional recovery measured by the BBB and IP tests following spinal cord injury. In addition, we provide a detailed neuroanatomical study of clip compression injury in mice that can be used to study the molecular mechanisms of SCI in knockout and transgenic mice. PMID- 11893023 TI - A mouse model of acute ischemic spinal cord injury. AB - Mice models of spinal cord injury (SCI) should improve our knowledge of the mechanisms of injury and repair of the nervous tissue. They represent a powerful tool for the development of therapeutic strategies in the fields of pharmacological, cellular, and genetic approaches of neurotrauma. We demonstrate here that the photochemical graded ischemic spinal cord injury model, described in rats, can be successfully adapted in mice, in a reliable and reproducible manner. Following the intravenous injection of Rose Bengal, the translucent dorsal surface of the T9 vertebral laminae of C57BL/6 female mice was irradiated with a 560-nm wavelength-light (3-8 min depending on the experimental group). Animals were sacrificed at 1 day or 7 days after injury. Functional tests were performed daily for motor, sensory, autonomic, and reflex responses. Lesion histopathology was assessed for lesion length, percentage of residual white matter, and astrocytic reactivity. Experimental groups demonstrated a functional deficit, which was correlated to the increase of the irradiation time and, therefore, to the severity of the injury. Histopathological and immunocytochemical data were reliable morphological measurements characterizing the degree of injury, which were strongly correlated to the severity of the functional impairment. Despite differences in the mechanism of injury, the wound healing response described in other traumatic SCI mice models was confirmed (no cavitation and, conversely, the formation of a dense connective tissue matrix). In this context, the precise understanding of the mechanisms of healing response after SCI in mice and of neurochemical kinetics appear to be crucial in the development of therapeutic strategies of CNS repair. Thus, the possible use of an increasing collection of transgenic mice offers a new dimension for experimental research in this area. The ischemic photochemical model of SCI in mice represents a relevant model that can play a key role in this new era of neurotrauma research. PMID- 11893025 TI - Compensatory locomotor adjustments of rats with cervical or thoracic spinal cord hemisections. AB - The accurate measurement of behavioral compensation after CNS trauma, such as spinal cord injury, is important when assessing the functional effects of injury and treatment in animal models. We investigated the locomotor abilities of rats with unilateral thoracic or cervical spinal cord injuries using a locomotor rating (BBB) scale, reflex tests, and quantitative kinetic measurements. The BBB rating scale indicated that thoracic spinal hemisected (TH) rats had more severely affected hindlimbs compared to cervical spinal hemisected (CH) and sham operated animals. Kinetic measurements revealed that CH and TH animals moved with different ground reaction force patterns, which nevertheless shared some similarities with each other and with the gait patterns of rats with different unilateral CNS lesions. Uninjured rats typically had an equal distribution of their body weight over the forelimbs and hind limbs, and used their forelimbs predominantly for braking while using their hind limbs mostly for propulsion. CH rats bore more weight on their hind limbs than their forelimbs, while TH animals bore more weight on their forelimbs than their hind limbs. Neither CH nor TH rats used the forelimb ipsilateral to the spinal hemisection for net braking or propulsion. The hindlimb contralateral to the hemisection was placed on the ground prematurely during the stride cycle for both CH and TH animals. The altered kinetics of the locomotor pattern in hemisected animals resulted in changes in the oscillations of total body potential and kinetic energies. These two forms of energy oscillate synchronously in intact locomoting rats, but were asynchronous during parts of the stride cycle in spinal hemisected animals. We conclude that rats develop a general compensatory response for unilateral CNS lesions, which may help stabilize the animal during locomotion. PMID- 11893024 TI - Intrathecal administration of epidermal growth factor and fibroblast growth factor 2 promotes ependymal proliferation and functional recovery after spinal cord injury in adult rats. AB - We have shown previously that epidermal growth factor (EGF) plus fibroblast growth factor (FGF2) expands the neural precursor cells in the ependyma of the normal adult rat spinal cord in vivo. To investigate the therapeutic effect of these factors on spinal cord injury (SCI), we administered EGF, FGF2, EGF plus FGF2, or artificial cerebrospinal fluid (aCSF) intrathecally (15 ng/h of EGF or FGF2) for 3 or 14 days after mild (2.4-g) or moderate (20-g) clip compression injury at T1 in adult rats. Histological and functional assessments were used to evaluate the therapeutic effects. The EGF plus FGF2 group, which received these agents for 14 days, showed better functional recovery than the aCSF group 42 days after moderate SCI (p < 0.05). At 14 days, the EGF plus FGF2 group showed a much greater expansion of ependymal cells and astrocytes compared to the other groups, and there was evidence for extensive migration of ependymal cells into the surrounding injured cord. These mitogens did not significantly enhance nestin expression in the ependymal layer or alter the expansion of oligodendrocyte precursor cells or microglia/macrophages, and dividing cells did not show the neuron-specific marker NeuN except immediately adjacent to the ependyma. The exact mechanism for improved functional recovery after EGF plus FGF2 is not known. PMID- 11893026 TI - Mechanisms of glutamate release in the rat spinal cord slices during metabolic inhibition. AB - Glutamate toxicity is a viable hypothesis to explain the expanding tissue degeneration occurring after traumatic or ischemic spinal cord injury. One important component in this process is the acute, excessive release of glutamate. In the current communication, the glycolytic inhibitor iodoacetate was used to induce metabolic inhibition in spinal cord slices and thereby provide an in vitro model to study the mechanisms of pathological glutamate release in the spinal cord. The evoked glutamate release was not Ca2+-dependent. Exclusion of NaCl reduced the evoked release of endogenous glutamate by 56%, while excluding Na+ increased release. Glutamate release was also reduced by the PLA2 inhibitors indomethacin (40%), arachidonyltrifluoromethyl ketone (45%) and 4-bromophenacyl bromide (36%). Blocking reverse glutamate transport by preincubation with 1 mM dihydrokainic acid reduced evoked release by 41%. However, when the dihydrokainic acid and arachidonyltrifluoromethyl ketone treatments were combined, no additive effect of the two substances was seen. These findings suggest that glutamate is released by three mechanisms from the energy compromised spinal cord: (1) in response to cellular swelling, most likely by the regulatory volume decrease, (2) by PLA2-mediated breakdown of the cell membrane and diffusion of glutamate down its concentration gradient, and (3) through reversal of the glutamate transporter. PMID- 11893027 TI - Locomotor performance of the rat after neonatal repairing of spinal cord injuries: quantitative assessment and electromyographic study. AB - In spinal cord injuries, various attempts have been made to reconstruct neural connections once disrupted. To improve current procedures and develop therapeutic methodologies it appears important to compare these reconstructive attempts via a standardized quantification of any ensuing functional recovery with parallel correlations to any potentially repaired neural connections. We have reported previously a quantitative assessment of neural connections across the graft site of rats whose spinal cord segments were neonatally replaced with embryonic spinal cord segments or a peripheral nerve section under comparable conditions. Using this same experimental model the present study assessed locomotor performance quantitatively using an open field locomotor scale at various postoperative intervals from day 0 to 5 weeks postinjury. To examine hind-forelimb coordination in further detail, electromyography was employed to record simultaneously from all four limbs during locomotion. Half of the rats whose spinal cord segments were repaired by replacement with embryonic homologous structures acquired virtually normal locomotor function, with a delay of five days compared with that of sham-operated rats. Detailed analysis revealed an abnormality in ankle joint movement and the stability of trunk during locomotion. Electromyography revealed that the pattern of locomotion in these rats was similar to controls. Grafted segments joined with the host spinal cord without gliosis at the host-graft interface. The remaining rats with an embryonic tissue graft showed various grades of hind-forelimb coordination. Gliosis and cavity were observed at the host-graft interface. The rats whose spinal cord was repaired by periphral nerve graft lacked hind-forelimb coordination despite the achievement of weight supported steps. It appears likely that the grade of locomotor performance depends on quantity and quality of reestablished neural connections across the graft. PMID- 11893028 TI - Amino acid concentrations in the blood of the jugular vein and peripheral artery after traumatic brain injury: decreased release of glutamate into the jugular vein in the early phase. AB - The gross behavior of excitatory amino acids in patients with traumatic brain injury (TBI), including uptake, transport, metabolism, and clearance, was investigated by analysis of the levels of 41 amino acids in the blood of the jugular vein (JV), which is the primary venous drainage conduit of the brain, and a peripheral artery. Blood samples from the JV and a peripheral artery of eight patients with TBI were collected at 6 h, 6 to 24 h, and over 24 h after TBI, and analyzed using high performance liquid chromatography. Blood samples from 101 normal subjects were also measured. The levels of glutamate (Glu), gamma aminobutyric acid (GABA), aspartate, glutamine, and cystine deviated from the normal range, and were considered pathological. The level of Glu in the JV was significantly lower than that in the artery (p < 0.05), and the level of GABA in the JV was significantly higher than that in the artery (p < 0.01), but the other three amino acids showed no significant differences. Significantly chronological changes in the difference between the blood levels in the JV and artery were observed for Glu. Measurement of the Glu level in the JV and artery may indicate gross metabolic change in the brain following TBI. PMID- 11893029 TI - Tissue microarrays: a powerful tool for high-throughput analysis of clinical specimens: a review of the method with validation data. AB - With recent advances in the knowledge of human molecular genetics, new gene-based disease mechanisms are emerging in many areas of medicine. The study of new prognostic and diagnostic markers in large numbers of clinical specimens is an important step in translating the new findings from basic science to clinical practice. The recently developed tissue microarray technology allows parallel molecular profiling of clinical samples at the DNA, RNA, and protein level. This technique enables pathologists to perform large-scale analyses using immunohistochemistry, fluorescence in situ hybridization, or RNA in situ hybridization substantially faster and at markedly lower costs compared with the conventional approach. This article provides a short review of this technology, focuses on several technical aspects of tissue microarray construction, and addresses the validity of the tissue microarray results for clinical research by reviewing data from recent literature along with the authors' own data. PMID- 11893030 TI - MIB-1 expression in cervical Papanicolaou tests correlates with dysplasia in subsequent cervical biopsies. AB - Ki-67 nuclear antigen is present in proliferating cells. MIB-1 antibody, raised to the recombinant part of the Ki-67 antigen, is a widely used biologic marker to assess cell proliferation. Ki-67 expression is normally observed in parabasal and basal cells in the cervix. With increasing severity of dysplasia, MIB-1 labeling is seen in cells of the superficial layers of cervical epithelium, which are exfoliated. The purpose of this study was to determine the sensitivity and specificity of presence of MIB-1-positive cells in Papanicolaou tests for predicting cervical neoplasia, condyloma, or both, on follow up. Using microwave antigen retrieval method, 49 air-dried cervical smears in two-smear cases were evaluated with immunostaining with MIB-1 monoclonal antibody. Presence of MIB-1 positivity was arbitrarily set at > or = 4 MIB-1 immunoreactive cells in each smear. The degree of positive staining was correlated with the cytologic diagnoses, subsequent colposcopy-directed biopsies, endocervical curettage, and/or cytologic follow ups. Follow-up findings correlated with cytology in 33 cases (67%), with MIB-1 positivity in 35 cases (71%). Three cases with positive follow ups were missed by cytology but detected by MIB-1 staining, and three cases were missed by MIB-1 but detected by cytology. Both cytology and MIB-1 staining failed to detect a subsequent cervical lesion in two cases, and in six cases each, an abnormal finding was not substantiated on follow ups. MIB-1 immunostaining is a powerful technique for evaluating gynecologic smears and is as equally sensitive and specific as cervical cytology. It is able to identify cervical disease overlooked by cytologic screening; therefore, it may serve as an adjunct and complimentary tool to cervical cytology. PMID- 11893031 TI - Evaluation of numeric alterations of chromosomes 1 and 17 by in situ hybridization in invasive breast carcinoma with clinicopathologic parameters. AB - Breast cancer is a genetically complex disease and is frequently associated with nonrandom chromosomal alterations. The occurrence of aberrations involving chromosomes 1 and 17 in malignant tissues of breast cancer patients has not been studied systematically. The numeric aberrations of chromosomes 1 and 17 were detected by nonisotopic in situ hybridization on paraffin-embedded tissue sections from 44 invasive breast carcinomas (42 cases available for chromosome 17) and were correlated with clinicopathologic parameters, patients' survival, p53, and c-erbB-2 proteins. Chromosome 17 and 1 aneuploidy were observed in the majority of breast carcinomas with equal percentages of polysomy and monosomy for chromosome 17 and predominance of polysomy for chromosome 1. Monosomy of chromosome 17 was significantly associated with positive lymph nodes and negative estrogen receptor (ER) immunohistochemical expression. Patients with chromosome 17 monosomy were at greater risk of death. Ductal carcinoma displayed a greater percentage of chromosome 1 polysomy than lobular ones. A statistically significant association was demonstrated between chromosome 1 polysomy and higher nuclear grade. Patients with chromosome 1 aneuploidy were at greater risk of death, and especially those with ER negativity. Aneuploid patients with c-erbB-2( )/PR(-) phenotype demonstrated lower survival rates. These data suggest a possible susceptibility of chromosome 17 to losses and gains and chromosome 1 to gains. Chromosome 17 monosomy and chromosome 1 aneuploidy may be useful prognostic markers in breast cancer patients. PMID- 11893032 TI - Immunocytochemical expression of epidermal growth factor receptor in myoepithelial cells of the breast. AB - The immunohistochemical expression and distribution of epidermal growth factor receptor (EGFr) in mammary myoepithelial cells (MECs) in normal tissue, benign epithelial proliferative lesions, and in situ carcinoma was performed. Results of the current study demonstrated that MECs stained constantly and strongly for EGFr, creating an outer continuous ring surrounding the epithelium of ducts and acini in healthy, in proliferative epithelial lesions, and in in situ carcinoma, both of ductal and lobular type. Foci of microinvasion were easily appreciated for the complete loss of EGFr immunostaining. Epidermal growth factor receptor expression in normal epithelia ranged from negative to weakly positive; it was positive in hyperplasia, whereas it was not constantly negative in in situ carcinoma. In conclusion, immunohistochemical staining for EGFr is diagnostically useful for MEC identification. The specific EGFr in MECs leads the authors to suggest that its expression may be related to the recently recognized high specialized paracrine function by which the MECs exert the natural mechanical and functional role in the juxtaposition between epithelium and stoma. PMID- 11893033 TI - CD44 associates with EGFR and erbB2 in metastasizing mammary carcinoma cells. AB - Type I receptor tyrosine kinases, including the epidermal growth factor receptor (EGFR) and erbB2, have been implicated in mammary carcinoma growth and metastasis. Recent evidence suggests that type I receptor signaling may be mediated by the CD44 family of transmembrane glycoproteins that also have been implicated in mammary tumor progression. Here, the authors tested whether CD44, EGFR, and erbB2 interacted and colocalized with one another in four mammary carcinoma cell lines (MCF-7, MDA-MB-231, MDA-MB-435, and MDA-MB-436) and in cytology samples obtained from patients with metastatic breast cancer. CD44 constitutively colocalized and coimmunoprecipitated with erbB2 and EGFR in all four mammary carcinoma cell lines. CD44 also colocalized with erbB2 and EGFR in all cytology samples expressing erbB2. CD44 colocalized with EGFR in cells from only 1 of 16 erbB2-negative cytology samples. These data indicate that CD44-EGFR erbB2 protein complexes occur in a high proportion of metastatic mammary carcinomas and suggest that CD44-type I receptor colocalization may be a novel prognostic marker for aggressive mammary cancers. PMID- 11893034 TI - Comparison of HER-2/neu oncogene amplification detected by fluorescence in situ hybridization in lobular and ductal breast cancer. AB - BACKGROUND: Abnormal expression of the HER-2/neu oncogene, a tyrosine kinase-type transmembrane growth factor receptor localized to chromosome 17q, has been associated with poor prognosis and the prediction of therapy response in invasive breast cancer. The comparative incidence and significance of HER-2/neu gene amplification for lobular and ductal breast cancer have not been previously characterized. DESIGN: Formalin-fixed, paraffin-embedded primary breast cancer tissue sections from 71 women diagnosed with invasive lobular carcinoma were tested for HER-2/neu gene amplification by fluorescence in situ hybridization (FISH) method using the Ventana unique sequence probe (Ventana Medical Systems, Tucson, AZ). A series of 106 cases of invasive ductal carcinoma was similarly processed and tested. Lymph node status was available for 155 (88%) of the 177 cases and 82 (46%) were lymph node-negative (LN-) and 73 (41%) were lymph node positive (LN+). Patients were treated for a mean of 65 months (range 1-169 months). RESULTS: 9 of 71 (13%) cases of lobular cancer featured HER-2/neu gene amplification, whereas 51 (48%) of 106 cases of ductal cancer showed amplification (P < 0.0001). On univariate analysis of combined lobular and ductal cases, HER-2/neu gene amplification detected by FISH predicted disease-related death (P < 0.0001). HER-2/neu gene amplification also predicted disease-related death in lobular cases alone (P = 0.003), LN+ lobular cases separately (P = 0.019), and LN- and LN+ ductal cases separately and alone (P < 0.0001). Multivariate analysis of the lobular group alone revealed that LN+ status (P = 0.015) and stage (P = 0.01) were independent predictors of disease-related death, and HER-2/neu gene amplification reached near significance (P = 0.086). In the ductal carcinoma group alone, HER-2/neu gene amplification (P = 0.03), lymph node status (P = 0.0001), tumor stage (P = 0.0001), and tumor grade (P = 0.044) were independent predictors of overall disease survival. CONCLUSIONS: HER-2/neu gene amplification detected by FISH was identified at a significantly lower rate in lobular compared with ductal breast cancer. HER-2/neu gene amplification when present in lobular breast cancer is a significant adverse prognostic factor. PMID- 11893035 TI - Role for microphthalmia transcription factor in the diagnosis of metastatic malignant melanoma. AB - Microphthalmia transcription factor (Mitf), a protein critical for the embryonic development and postnatal viability of melanocytes, is a master lineage regulator and modulates extracellular signals. Recently, an anti-Mitf antibody, D5, was shown to be both a sensitive and a specific marker of epithelioid melanoma. Those data suggested that Mitf expression was specific for melanocytic differentiation because it was not detected in six different carcinoma types. To broaden the spectrum of melanomatous and nonmelanomatous tumors tested with this antibody, the authors investigated the sensitivity and specificity of the D5 anti-Mitf antibody in 36 metastatic melanomas and 102 nonmelanomatous tumors. Twenty-nine of 36 (81%) melanomas examined were reactive for D5. Of these, 29 of 32 (91%) epithelioid melanomas were reactive, whereas all 4 melanomas with spindle cell morphology were nonreactive. Six of 102 (5.8%) nonmelanomatous tumors (1 breast carcinoma, 1 malignant mixed mullerian tumor, 2 renal cell carcinomas, and 2 leiomyosarcomas) were reactive for D5. All melanomas were reactive for S-100 protein (a criteria for inclusion in the study), whereas 8 of 102 (7.8%) of nonmelanomatous tumors were reactive. Twenty-five of 36 melanomas (69%) were reactive for HMB-45, whereas no nonmelanomatous tumors were reactive. Twenty-one of 28 (75%) melanomas were reactive for Melan-A. Reactivity for Melan-A was detected in 5 of 102 (4.9%) nonmelanomatous tumors. As with D5, HMB-45 and Melan A failed to detect all spindle-cell melanomas. Results of this study confirm that D5 is a sensitive marker for epithelioid melanomas; however, the authors found D5 neither quite as sensitive nor quite as specific as in previous studies. Nevertheless, the authors believe that positive staining for D5 when taken in clinical, histologic, and immunohistochemical context may be diagnostically useful. In particular, D5 is helpful in the evaluation of epithelioid neoplasms, but not spindle-cell tumors. PMID- 11893036 TI - Immunocytochemical increased evidence of inducible nitric oxide synthase, tumor necrosis factor-alpha, and adrenocorticotropic hormone in human peritumoral lymph nodes. AB - In the current study, mesenteric and peritumoral lymph nodes surgically removed from patients with colon-rectum cancer were studied. Morphologic and immunocytochemical investigations demonstrated that mesenteric (control) and peritumoral lymph nodes of a same patient showed the same morphologic structure, but a different immunocytochemical pattern. Indeed, an increased immunoreactivity to anti-inducible nitric oxide synthase, anti-tumor necrosis factor-alpha, and anti-adrenocorticotropic hormone antibodies in the lymphatic tissue of peritumoral lymph nodes compared with mesenteric lymph nodes was observed. These findings suggest that in colon-rectum cancer, the pathologic event induces an increased expression of the molecules involved in the processes of inflammation and carcinogenesis that occurs earlier than the appearance of morphologic modifications. PMID- 11893037 TI - Immunohistochemical screening of mismatch repair genes hMLH1, hMSH2, and hMSH6 in dysplastic lesions of the colon. AB - Immunohistochemical decrease in staining for mismatch repair proteins may be seen in either microsatellite instability or inactivation (methylation) of mismatch repair proteins. Both are features of the malignant phenotype in a range of colorectal neoplasms. Expression of mismatch repair proteins in dysplastic lesions in ulcerative colitis (UC) has not been studied extensively. The authors analyzed protein expression of hMLH1, hMSH2, and hMSH6 in 18 cases of dysplasia associated lesion or mass (DALM) in patients with UC and 10 sporadic adenomas. Immunohistochemical studies revealed adequate staining in almost all of the cases. A significant decrease in protein expression was seen in 2 DALM and 2 sporadic adenomas. The authors conclude that immunohistochemical studies of mismatch repair proteins can be applied to dysplastic lesions in UC with adequate staining results. Decreased wild type expression of hMLH1, hMSH2, and hMSH6 is not a feature of DALM in the setting of UC. PMID- 11893038 TI - Expression of maspin in pancreatic neoplasms: application of maspin immunohistochemistry to the differential diagnosis. AB - Maspin is a unique member of the serpin family, which inhibits tumor invasion and metastasis of the human breast and prostate cancers. Immunohistochemical evaluation of the maspin expression in pancreatic neoplasms has never been performed. The authors examined the expression of maspin in various pancreatic neoplasms to investigate its usefulness as an adjunct diagnostic marker. Formalin fixed, paraffin-embedded tissue sections from 107 pancreatic neoplasms were immunostained with anti-maspin antibody using an EnVision+ System. Maspin was expressed in all ductal adenocarcinomas, of which 78.9% (30/38) were high expressors and 21.1% (8/38) were low expressors. All 13 intraductal papillary mucinous tumors and 11 of the 13 mucinous cystic tumors reacted to maspin with stronger expressions in carcinomatous lesions. In contrast, acinar cell carcinoma, pancreatic endocrine tumors, solid-pseudopapillary tumors, and serous cystadenomas demonstrated no maspin expression. In addition, nonneoplastic pancreatic parenchyma and chronic pancreatitis lacked expression. The current study suggests that maspin may be of importance in the pathobiology of pancreatic neoplasms with epithelial origin, especially pancreatic tumors that are composed of mucin-producing cells. It may be useful in separating ductal adenocarcinoma from acinar cell carcinoma, pancreatic endocrine tumor, solid-pseudopapillary tumor, and chronic pancreatitis, especially in needle biopsy specimens. PMID- 11893039 TI - Calretinin and inhibin are useful in separating adrenocortical neoplasms from pheochromocytomas. AB - Most adrenocortical neoplasms and pheochromocytomas can be diagnosed by a combination of clinical findings and morphologic features. Occasionally, however, this histologic differential diagnosis requires ancillary tests, such as immunohistochemistry. Both tumors are generally negative for epithelial markers but express synaptophysin. Inhibin and chromogranin are used for the diagnosis of adrenocortical neoplasms and pheochromocytomas, respectively. Both antigens, however, are expressed focally and may be completely negative, particularly in small biopsies. The authors investigated the potential value of adding calretinin to inhibin in the differential diagnosis of these tumors. Fifty-five primary adrenal neoplasms including 33 adrenocortical tumors (21 adenomas and 12 carcinomas), 22 pheochromocytomas, and 7 healthy adrenal glands were examined immunohistochemically for the expression of calretinin and inhibin. Inhibin was demonstrated in 24 (73%) adrenocortical neoplasms. When calretinin was added, the number of tumors staining positively for the two markers alone or in combination increased to 31 (94%). Both antigens showed a focal pattern of distribution in many cases. None of the pheochromocytomas reacted for any of these two markers. Healthy adrenal gland showed a distinct positive and negative pattern of immunoreactivity for both antigens in cortex and medulla, respectively. There were no differences between staining patterns of calretinin and inhibin in healthy adrenal cortex, adrenocortical adenomas, and adrenocortical carcinomas. The authors conclude that the addition of calretinin to inhibin increases the sensitivity of the diagnosis of adrenocortical neoplasms. When used together, they are highly specific and sensitive for the differential diagnosis of these tumors from pheochromocytomas. These markers, however, do not distinguish between benign and malignant adrenocortical neoplasms. PMID- 11893040 TI - Hydroa-like cutaneous T-cell lymphoma: a clinicopathologic and molecular genetic study of 16 pediatric cases from Peru. AB - Hydroa-like cutaneous T-cell lymphoma (hydroa-like CTCL) is an unusual pediatric malignancy with a poor prognosis. An impressive cutaneous rash characterized by edema, blisters, ulcers, crusts, and scars, resembling hidroa vacciniforme, is seen mainly on the face and sometimes on the extremities. The lesion consists of lymphomatous T-cell infiltration of the skin and subcutis with variable exocytosis and angiocentricity. It has been also called edematous, scarring vasculitic panniculitis and hydroa-like lymphoma. An association with Epstein Barr virus has been suggested. The differential diagnosis includes other cutaneous lymphomas, particularly the cutaneous nasal type T/natural killer-cell lymphoma, mycosis fungoides, precursor T-cell lymphoblastic lymphoma, nonspecific peripheral T-cell lymphoma, cutaneous anaplastic large cell lymphoma, and subcutaneous panniculitic T-cell lymphoma. Other differential diagnoses are inflammatory dermatopathies and panniculitides. Based on a series of 16 such cases referred to the Institute of Neoplastic Diseases, the objective of this report is not only to provide a better clinicopathologic understanding of this entity but also a reappraisal of it as a malignancy. The male/female frequency ratio was 1:1. The median age was 10 years old. All cases showed predominant facial involvement with edema, blisters, ulcers, crusts, and scars. Chemotherapy and/or radiotherapy had little or no benefit. The prognosis was usually dismal. The lymphoma extended from the epidermis to the subcutis, with frequent angiocentric and periadnexal array. Lymphoma cells were mostly of intermediate size with dense hyperchromatic nuclei, inconspicuous nucleoli, and infrequent mitosis. A scanty and variable inflammatory background was found. The lymphoma cells displayed T-cell cytotoxic phenotype. In addition, they were negative for the natural killer cell antigens CD56 and CD57. Epstein-Barr virus in situ hybridization was positive in the six cases in which it was assayed. T-cell receptor gamma (TCRgamma) displayed monoclonal-type rearrangement in four cases studied. Our findings indicate that hydroa-like CTCL is an independent clinicopathologic entity that affects children. Consequently, it should be considered an independent subset of CTCLs and be included as such in the classification of neoplastic diseases of the lymphoid tissues. PMID- 11893041 TI - Morphologic, immunohistochemical, immunologic, ultrastructural, and time-related study of herpes simplex virus type 1-infected cultured human fibroblasts. AB - Membrane glycoproteins of enveloped animal viruses are synthesized, processed, and transported inside infected cells. Expression of viral glycoproteins on the surface of viral particles and host cells are essential for many biologic functions. In the case of herpes simplex virus, the glycoprotein molecules may act as nucleation points for virus assembly and budding at the nuclear membrane. The temporal distribution of herpes simplex virus type 1 particles and glycoproteins in cultured human fibroblasts was studied by titration plaque assay, immunoblots, immunofluorescence light microscopy, and immunogold cryosection electron microscopy to describe the virus-cell interactions. These concordant analyses revealed significant release of infectious viral particles to the medium at 6 hours postinfection, that the capacity of the host cells to make infectious viral particles was complete at 18 hours postinfection, and that the infection brought time-related modifications of tubulin, cell morphology, and viral glycoproteins. The data presented is in accord with the theory of envelopment at the nuclear membranes containing immature glycoproteins followed by multiple deenvelopments and reenvelopments of the virus particles during the transport and maturation in the endoplasmic reticulum and the Golgi complex. PMID- 11893042 TI - Is immunostaining for Helicobacter pylori superior to the special stain thiazine in detecting small numbers of H. pylori in gastric biopsies? AB - Helicobacter pylori is implicated in the pathogenesis of gastritis and duodenal ulcers, gastric lymphoma of the mucosa-associated lymphoid tissue (MALT) type, and gastric adenocarcinoma. Eradication of H. pylori with antibiotic therapy therefore is essential, not only for the successful treatment of active gastritis, but also for the treatment and prevention of the MALT lymphoma. It has been suggested recently that immunostaining for H. pylori is more sensitive than special stains for the detection of the organism in the gastric biopsies after triple therapy. Fifty-five endoscopic mucosal biopsies from 38 patients, including 18 treated with H. pylori eradication therapy, were selected for immunostaining because they were either negative or contained rare H. pylori organisms by thiazine stain. Formalin-fixed, paraffin-embedded tissue sections were immunostained for H. pylori using a polyclonal antibody using standard immunoperoxidase technique. The results were compared with those obtained with the thiazine stain. Detection of H. pylori by immunostaining was easier and less time-consuming than by thiazine stain. There was complete agreement between immunostaining and thiazine stain in 48 (87%) cases. Of the 7 discordant cases, 3 (42%) were positive for H. pylori with thiazine only and 4 (48%) with immunostaining only. Given the nature of the selection of the study sample (absent to rare by thiazine stain), the discordance most likely represents a sampling error. The authors concluded that immunostaining for H. pylori did not appear to be more sensitive than special stains. Three cases with bacterial clumps were diagnosed previously as positive for H. pylori, but identified correctly as negative using both staining methods. Pathologists, however, should balance the added cost to patients of immunostaining against the time saved by the easier screening of the immunostained slides and the possibility of false positive results when special stains are interpreted by inexperienced pathologists. PMID- 11893043 TI - Double-label confocal laser-scanning microscopy, image restoration, and real-time three-dimensional reconstruction to study axons in the central nervous system and their contacts with target neurons. AB - The current double tracing-double confocal laser-scanning method was developed to reconstruct identified nerve fibers and their contacts with identified target neurons in the rat brain in three dimensions. It intends to fill the gap between conventional light microscopic and electron microscopic neuroanatomic tracing. The steps involved are as follows: (1) injection of two neuroanatomic tracers- Phaseolus vulgaris leucoagglutinin (PHA-L) to label fibers innervating a particular brain area and Neurobiotin to label prospective target neurons in that area; (2) immunofluorescence detection of the labeled fibers (fluorophore Cy5, infrared emission), together with fluorochromated avidin detection of the taken up Neurobiotin (Cy2 or Alexa 488; green emission); (3) acquisition of Z-series of confocal images at high magnification with a laser-scanning microscope using the laser lines 488 nm and 647 nm; and (4) computer-processing and three-dimensional reconstruction of the labeled fibers and the presumed target dendrites. Rotation on the computer of the three-dimensional reconstructed fibers and dendrites along all three spatial axes enabled the authors to determine whether "true" or "false" contacts occur. In a true contact no space was present between the apposing structures, whereas a false contact consisted of two differently stained structures close to each other but separated by a narrow, optically empty space. One important phenomenon in the three-dimensional reconstruction of double stained structures that needed correction was "twin image mismatch"--i.e., the observation that a three-dimensional reconstruction of a small test object (double-stained on purpose) produced two slightly shifted objects, each associated with its particular fluorochrome. To measure the actual twin image mismatch of the confocal instrument and to obtain accurate correction factors the authors took in each session in which they obtained image series of the real experiments, with both laser wavelengths Z-series of images of multifluorescent microspheres (500-nm diameter) and of thin, double-stained fibers. Given the small dimensions of the structures of interest, i.e., synaptic contacts, it is necessary in this type of research that the optical characteristics of the imaging system--e.g., the alignment errors and chromatic aberration that produce twin image mismatch--be precisely known. PMID- 11893044 TI - Nonvital teeth: a list of questions. PMID- 11893045 TI - Reliability of in vitro microleakage tests: a literature review. AB - PURPOSE: The literature contains conflicting data about in vitro microleakage evaluations and their usefulness and reliability. No standardization has yet been established. Here we consider features of published studies that might affect the results of the in vitro microleakage tests. MATERIALS AND METHODS: We reviewed 144 in vitro microleakage studies, published in 14 international reviews between 1992 and 1998, which comprised 917 sets or groups of experiments. The published studies were entered in a database and compared using selected literature criteria: sample, cavities, restoration procedures, thermocycling and mechanical cycling, evaluation method. RESULTS: The methods employed vary widely. The most frequent methodological choices (%) were (1) specimen storage after extraction: duration (unspecified, 59.2), medium (distilled or deionized water, 33.8), temperature (unspecified, 52.2), additives (none, 47.0); (2) aging method (79.1): duration before aging (< 24 h, 35.9); medium and temperature of storage before aging (distilled or deionized water, 26.8; 37 degrees C, 54.3); (3) medium of cycling (tap water, 50.5), number of cycles ([250-500], 34.6), number of baths (2, 84.0), bath temperature (5 degrees C to 55 degrees C, 60.6), immersion dwell time (30 s, 44.3); (4) tracer: type (basic fuchsin, 40.7), time of immersion (after thermocycling and/or mechanical cycles, 64.1), immersion duration (basic fuchsin: 24 h, 59.5); assessment of dye penetration of sections (91.7): direction (perpendicular, 88.5), number (1, 47.1). CONCLUSION: The great variability in the methods used in these 144 studies prevented meta-analysis and comparison of the results, thus reducing the value of these methods. PMID- 11893046 TI - Tensile strength of human dentin as a function of tubule orientation and density. AB - PURPOSE: This study investigated the ultimate tensile strength (UTS) of human dentin as a function of tubule orientation and density. MATERIALS AND METHODS: Slabs of dentin (ca 0.7 mm thick) were obtained from human third molars by either transversally or longitudinally sectioning the crowns with an Isomet saw. The slabs were gently trimmed to reduce the central area of the coronal dentin to a cross-sectional area of approximately 0.5 mm2. The longitudinally sectioned slabs were either trimmed from the mesial and distal sides or from the occlusal and pulpal aspects to permit the tensile load to be applied either parallel or perpendicular to the tubule orientation. The transversally sectioned specimens were obtained at several distances from the pulp and were used to evaluate the effects of tubule density. The trimmed specimens were tested in tension on a Kratos testing machine at 0.5 mm/min. After failure, the UTS of each was calculated and expressed in MPa. The fractured ends of the transversally sectioned specimens were viewed under SEM to calculate the number of tubules per mm2 at the site of fracture, and its relation with the UTS was investigated by regression analysis. RESULTS: The UTS of dentin is higher when the load is applied perpendicular to the tubule orientation (80 +/- 13 MPa) than when applied parallel to tubule orientation (58 +/- 11 MPa, p < 0.05). There was a tendency for dentin to be weaker as the number of tubules at the site of fracture increased, although this relationship was not statistically significant (R2 = 0.051, p > 0.05). CONCLUSION: The UTS of dentin is dependent on the tubule direction. Dentin tends to be weaker as the number of tubules per area increases. PMID- 11893047 TI - The influence of tubule density and area of solid dentin on bond strength of two adhesive systems to dentin. AB - PURPOSE: To determine the correlation between the tubule density (TD) and the area occupied by solid dentin (ASD) with the bond strength of one conventional and one self-etching adhesive system to dentin. MATERIALS AND METHODS: The crown of extracted human third molars was transversally sectioned with a diamond saw to expose either superficial, middle, or deep dentin. The three groups of dentin surfaces were randomly divided and bonded with either Clearfil Liner Bond 2V (LB) or Prime & Bond 2.1 (PB) adhesive systems according to manufacturer's directions. Resin composite buildup crowns (10.0 mm high) were incrementally constructed on the bonded surfaces and the teeth stored in water at 37 degrees C. After 24 h of storage, the teeth were vertically, serially sectioned in both x and y directions to obtain several bonded sticks of approximately 0.7 mm2 cross-sectional area. Each stick was tested in tension in a EMIC DL-500 tester at 0.5 mm/min until failure. After testing, the dentin side of the fractured specimen was gently abraded with a 1000-grit SiC paper, etched with 37% phosphoric acid for 15 s and allowed to air dry. SEM micrographs at 1000x and 4000x magnification were taken to permit calculation of the TD (number of tubules/mm2) and ASD (% of total area) at the site of fracture. Correlation between TD and ASD with the bond strength data was performed by linear regression. All statistical analysis was done with alpha = 0.05. RESULTS: Overall bond strength (MPa) for LB was 26.0 +/- 10.2, and 42.6 +/- 15.2 for PB. There was a significant direct relationship between bond strength and ASD for both materials (r2 = 0.20, p < 0.05 and r2 = 0.66, p < 0.01, respectively for LB and PB). PB bond strength dropped significantly as the TD increased (r2 = 0.63, p < 0.05), while LB was not sensitive to TD (r2 = 0.05, p > 0.05). Mean bond strength of PB was significantly higher than LB for both superficial and middle dentin (p < 0.05), while there was no significant difference for deep dentin (p > 0.05). CONCLUSION: Regional variations in TD and ASD may modify bond strength of both conventional and self-etching adhesive systems. Bonding sites with larger ASD seem to yield higher bond strengths regardless of the type of adhesive system used. PMID- 11893048 TI - Effect of dentin conditioners on wet bonding of 4-META/MMA-TBB resin. AB - PURPOSE: By altering either ferric chloride concentration in 10% citric acid (1% ferric chloride = 10-1; 5% ferric chloride = 10-5; 10% ferric chloride = 10-10) or conditioning periods with an aqueous mixture of 1% citric acid and 1% ferric chloride (1-1), the influence of dentin substrate on bond strength and hybridized dentin in wet bonding of 4-META/MMA-TBB resin was examined. MATERIALS AND METHODS: Dentin surfaces of fresh bovine incisors were conditioned either with 10 1, 10-5, or 10-10 mixtures for 10 s (10-1-10s, 10-5-10s, 10-10-10s groups) or with a 1-1 mixture for 5, 10, 30 or 60 s (1-1-5s, 1-1-10s, 1-1-30s, 1-1-60s groups). Rinsed, demineralized dentin samples were kept wet, primed with 5% 4 META in acetone for 60 s, and bonded with 4-META/MMA-TBB resin. Bonded specimens were trimmed to a mini-dumbbell shape for tensile testing. The cross sections of bonded specimens were modified with HCl and NaOCl in order to assess the hybrid layer. The fractured surfaces of specimens and the hybridized dentin were investigated with SEM. RESULTS: No significant difference (p > 0.01) in tensile strength was identified between 10-1-10s and 10-5-10s groups (30 MPa), 10-10-10s and 1-1-5s groups (15 MPa), and the three groups conditioned by 1-1-10s, -30s and -60s (40 MPa). The thickness of the hybrid layer increased with increasing either ferric chloride or conditioning periods. CONCLUSION: The concentration of ferric chloride in 10% citric acid for wet bonding must be less than 5% in order to provide a reliable bond. When applied from 10 to 60 s, the 1-1 conditioner provided hybridized dentin with reliable tensile bond strength. The thickness of the hybrid layer did not influence the tensile bond strength. PMID- 11893049 TI - Microtensile bond strength evaluation of three adhesive systems in cervical dentin cavities. AB - PURPOSE: To evaluate the influence of the orientation of dentinal tubules on the bonding performance, microtensile bond strengths (MTBS) to dentin in cervical dentin cavities were measured for three current bonding systems. MATERIALS AND METHODS: Wedge-shaped cervical cavities (5 x 5 x 3 mm) prepared in extracted human premolars were treated with one of the three adhesive systems, Gluma One Bond, UniFil Bond and Mega Bond in combination with a hybrid-type resin composite. After storage in water for 24 h, the teeth were sectioned longitudinally through the restoration for determination of MTBS, either at the coronal or at the apical wall of the lesion. In an additional group, either the coronal or the apical walls were coated with vaseline prior to adhesive bonding and insertion of the resin composite, and then MTBS was measured. RESULTS: MTBSs (mean +/- SD, MPa) to coronal and apical dentin were 35.1 +/- 19.1 and 16.4 +/- 7.9 for Gluma One Bond, 37.7 +/- 11.5 and 18.6 +/- 8.6 for UniFil Bond, and 27.0 +/- 15.2 and 30.3 +/- 17.0 for Mega Bond, respectively. MTBS to the coronal wall was higher than to the apical wall (p < 0.05) with Gluma One Bond and UniFil Bond, whereas no difference was found with Mega Bond. With all three systems, vaseline coating had no effect on the bond strength (p > 0.05), indicating that the wall-to-wall contraction stresses exerted in the noncoated group had no influence on the bond strength generated. CONCLUSION: In cervical dentin cavities, apart from the individual adhesive's bonding capacity, the dentinal tubular orientation may have an influence on the bond strength. PMID- 11893050 TI - Six-month clinical evaluation of two dentin adhesives applied on dry vs moist dentin. AB - PURPOSE: Acid-etched dentin has been described to easily collapse when it is dried with air after being rinsed with water under laboratorial "ideal" conditions. Manufacturers of modern dentin adhesives recommend leaving dentin moist, regardless of the solvent used in their proprietary dentin adhesives. Nevertheless, many clinicians still dry the cavity preparation after rinsing off the etching gel to check for the enamel frosted aspect. The null hypothesis to be tested in this clinical study was that drying dentin with air upon rinsing off the acid would not result in lower short-term retention rates than when the cavity was left visibly moist. MATERIALS AND METHODS: Upon Internal Review Board approval, thirty-five patients were enrolled in this study. A total of 128 restorations divided into four groups were inserted and evaluated at baseline: (A) Prime & Bond NT, an acetone-based adhesive, applied on moist dentin; (B) Prime & Bond NT applied on dentin dried with air for 3 to 4 s; (C) Single Bond, an ethanol- and water-based adhesive, applied on moist dentin; (D) Single Bond applied on dentin dried with air for 3 to 4 s. A microfilled composite resin was used for all restorations. RESULTS: At 6 months after initial placement, 119 restorations (a 93% recall rate) were re-evaluated. Retention rates at 6 months were 97% (one failure) for Single Bond/moist dentin and 100% for the remaining three groups; however, they were not significantly different (retention rate vs dentin adhesive; retention rate vs moisture). CONCLUSION: Dentin substrate in noncarious lesions may be less sensitive to variations in dentin moisture than the "ideal" dentin substrate used under laboratory conditions. PMID- 11893051 TI - Clinical procedure for luting glass-fiber posts. AB - PURPOSE: This report presents clinical cases in which a self-activating dual-cure adhesive system was used in combination with proprietary self-curing resin cement for bonding a translucent glass-fiber post. The clinical luting procedure of a self-activating bonding/self-curing resin cement system in combination with a glass-fiber post based on fiber-reinforced technology is documented. MATERIALS AND METHODS: The glass-fiber post was placed into a root canal preparation under clinical conditions and documented. A few restored teeth were extracted after 1 week and processed for SEM observations to determine the bonding mechanism to root dentin under clinical conditions. RESULTS: The SEM observations clearly showed that the bonding mechanism between Excite Dual Cure and root dentin was based on hybrid layer, resin tag, and adhesive lateral branch formation. CONCLUSION: The luting procedure described is simple and easy, and can be proposed as a daily clinical technique. PMID- 11893052 TI - Test methods to evaluate adhesive strength to tooth structure. PMID- 11893053 TI - Factors influencing the reductive cleavage of C-x multiple bonds in their reactions with meal-meal multiple bonds (X = C, N, O, S). AB - Though metal-metal multiple bonds of the transition elements are redox active, their reactivity towards C-X multiple bonds (X = C, N, O, S) vary greatly depending principally on: 1. The coordination geometry of the metal. 2. The oxidation state of the metal and the electronic configuration of the M-M bond. 3. The nature of the attendant ligands. Specific examples of C-X multiple bond activation at dimolybdenum and ditungsten centers are presented that illustrate the importance of these factors. Evidence is presented to support the view that reductive cleavage of a C-X multiple bond can be considered to be equivalent to an intramolecular redox reaction within a [M2CX] "cluster complex," for which the frontier orbital energies of the C-X and M-M multiple bonds are of paramount importance. Some applications of these C-X reductive cleavage reactions toward organic synthesis are described. PMID- 11893054 TI - The Clostridium cellulovorans cellulosome: an enzyme complex with plant cell wall degrading activity. AB - Cellulose comprises a major portion of biomass on the earth, and the turnover of this material contributes to the CO2 cycle. Cellulases, which play a major role in the turnover of cellulosic materials, have been found either as free enzymes that work synergistically, or as an enzyme complex called the cellulosome. This review summarizes some of the general properties of cellulosomes, and more specifically, the properties of the Clostridium cellulovorans cellulosome. The C cellulovorans cellulosome is an extracellular enzyme complex with a molecular weight of about 1 x 10(6), and is comprised of at least ten subunits. The major subunit is the scaffolding protein CbpA, with a molecular weight of 189,000. This nonenzymatic subunit contains a cellulose binding domain (CBD) that binds the cellulosome to the substrate, nine conserved cohesins or enzyme binding domains, and four conserved surface layer homologous (SLH) domains. It is postulated that the SLH domains help to bind the cellulosome to the cell surface. The cellulosomal enzymes include cellulases (family 5 and 9 endoglucanases and a family 48 exoglucanase), a mannanase, a xylanase, and a pectate lyase. The cellulosome is capable of converting Arabidopsis and tobacco plant cells to protoplasts. One of the endoglucanases, EngE, contains three tandemly repeated SLHs at its N-terminus, and therefore appears capable of binding to the scaffolding protein CbpA as well as to the cell surface. Cellulosomes can attack crystalline cellulose, but the free cellulosomal enzymes can attack only soluble and amorphous celluloses. Nine genes for the cellulosome are found in a gene cluster cbpA-exgS-engH-engK-hbpA-engL-manA-engM-engN. Other cellulosomal genes such as engB, engE, and engY are not linked to the major gene cluster or to each other. By determining the structure and function of the cellulosome, we hope to increase the efficiency of the cellulosome by genetic engineering techniques. PMID- 11893055 TI - Biomimetic selectivity. AB - Synthetic organic chemistry normally achieves selectivity by manipulation of the intrinsic reactivity of the substrate, but enzyme use is quite a different principle. The geometry of the enzyme-substrate complex determines enzymatic selectivity, completely overwhelming any normal selective reactivities. Biomimetic chemistry aims to imitate the enzymatic style. Some early approaches used attached reagents or templates to direct photochemical and free radical processes, with a combination of geometric and reactivity control. Recent work uses a mimic of the enzyme class cytochrome P-450 to achieve the selective hydroxylations of steroids with complete domination by the geometry of the catalyst-substrate complex. PMID- 11893056 TI - Heterogeneous catalysis on the atomic scale. AB - Information about the elementary processes underlying heterogeneous catalysis may be obtained by investigating well-defined single crystal surfaces. The success of this "surface science" approach for "'real" catalysis can be demonstrated, for example, with ammonia synthesis. The progress of catalytic reactions can be observed on an atomic scale by applying scanning tunneling microscopy and other surface physical techniques, as is shown with different examples in this paper: CO oxidation on a Pt(111) surface proceeds preferentially along the boundaries between adsorbed O and CO patches. Ru is practically inactive for the same reaction under lower pressure conditions but is transformed into RuO2 under atmospheric pressure conditions, where part of the surface Ru atoms function as coordinatively unsaturated sites (cus). In contrast, in the hydrogen oxidation reaction on Pt(111), an autocatalytic reaction step comes into prominence, and is responsible for the formation of propagating concentration patterns on the surface as a characteristic of nonlinear dynamics. Additionally, the limits of the concept of thermal equilibrium in surface rate processes are explored by applying ultrafast (femtosecond) laser techniques. PMID- 11893057 TI - Stereocontrol in radical polymerization. AB - The stereospecific radical polymerization of vinyl esters, methacrylates, and alpha-substituted acrylates was studied. Fluoroalcohols, as a solvent, have remarkable effects on the stereoregularity of the radical polymerizations of vinyl acetate, vinyl pivalate, and vinyl benzoate, affording polymers rich in syndiotacticity, heterotacticity, and isotacticity, respectively. This method was successfully applied to the polymerization of methacrylates to give syndiotactic polymers. The steric repulsion between the entering monomer and the chain-end monomeric unit bound by the solvent through hydrogen bonding is important for the stereochemical control in these systems. Lewis acid catalysts, such as lanthanide trifluoromethanesulfonates and zinc salts, were also effective for the stereocontrol during the radical polymerization of methyl methacrylate, to reduce the syndiotacticity and alpha-(alkoxymethyl)acrylates to synthesize isotactic and syndiotactic polymers. Radical polymerization of the methacrylates bearing a bulky ester group, such as the triphenylmethyl methacrylate derivatives, gave highly isotactic polymers, as in the case of anionic polymerization. In addition, the control of one-handed helical conformation was attained in the radical polymerization of 1-phenyldibenzosuberyl methacrylate using chiral neomenthanethiol or cobalt(II) complexes as an additive. PMID- 11893058 TI - Template-directed ligation: from DNA towards different versatile templates. AB - A systematic approach evaluating template-directed ligation reactions has now resulted in a simple outline for a two-stage replication cycle. This cycle builds on an efficient method for reading the information encoded in DNA into an amplified translation product. It is further demonstrated that the translation product strand is capable of catalyzing the synthesis of the original DNA strand. We propose that this cycle represents just one of many possible solutions; other chemical ligation or polymerization reactions could be accommodated with different templates. In that context, a new template, derived by modest changes to the DNA backbone, has been developed and has been shown to hybridize under reaction conditions different than those accessible to DNA. Therefore, the conceptual groundwork has been laid for extending this approach to encoding and reading stored information in molecules other than the natural biopolymers at the densities found in biology. PMID- 11893059 TI - Donepezil hydrochloride: a treatment drug for Alzheimer's disease. AB - The role of the cholinergic system with respect to cognitive deficits characteristic of Alzheimer's disease (AD) has led to a number of studies focusing on the development of acetylcholinesterase (AChE) inhibitors as a drug for treating this disease. The earliest known AChE inhibitors, namely, physostigmine and tacrine, performed poorly in clinical trials (e.g., poor oral activity, brain penetration, and hepatotoxic liability). Studies were then focused on finding a new type of acetylcholinesterase inhibitor that would overcome the disadvantages of these two compounds. Donepezil hydrochloride inaugurates a new class of AChE inhibitors with longer and more selective action and with manageable adverse effects. PMID- 11893060 TI - Cofactor diversity in biological oxidations: implications and applications. AB - Until recently, it was generally believed that enzymatic oxidation and reduction requires the participation of either a nicotinamide (NAD(P)+) or a flavin (FAD, FMN), in agreement with the existence of NAD(P)/H-dependent dehydrogenases/reductases and flavoprotein dehydrogenases/reductases/oxidases. However, during the past 20 years, the unraveling of the enzymology of the oxidation and reduction of C1-compounds by bacteria has led to the discovery of many new redox cofactors, some of them discussed here as they have a wider physiological significance than just enabling enzymatic C1-conversions to occur. A good example is the quinone cofactors, encompassing PQQ (2,7,9-tricarboxy-1H pyrrolo[2,3-f]-quinoline-4,5-dione), TTQ (tryptophyl tryptophanquinone), TPQ (topaquinone), LTQ (lysyl topaquinone), and several others whose structures have still to be elucidated. Another example is mycothiol (1-O-(2'-[N-acetyl-L cysteinyl]amido-2'-deoxy-alpha-D-glucopyranosyl)-D-myo-inosoitol), the counterpart of glutathione, once thought to be a universal coenzyme. Because these novel cofactors assist in reactions that can also be catalyzed by already known enzyme "classic cofactor" combinations, and first indications suggest that the chemistry of the reactions is not unique, one may wonder about the evolutionary background for this cofactor diversity. However, as will be illustrated by examples, from a practical point of view the diversity is beneficial, as it has increased the arsenal of enzymes suitable for application. PMID- 11893061 TI - Polymer surface science. AB - Molecular level studies of the structure and mechanical properties of polymer surfaces have been carried out by sum frequency generation (SFG) surface vibrational spectroscopy and atomic force microscopy (AFM). The surfaces of different grades of polyethylene and polypropylene have been characterized including during the glass transition and when mechanically stretched. Copolymers that have hard and soft segments with different glass transition temperatures show phase separation, an effect of hydrogen bonding between the hard and soft segments, that influences their adhesive and friction properties. AFM and SFG show that low surface energy additives migrate to the surface and alter the surface mechanical properties. Polymers, where the chemical nature of the end groups is different from the backbone, show surface segregation of the hydrophobic part of the chain in air and the hydrophilic part in water. Likewise, in miscible polymer blends, surface segregation of the more hydrophobic component in air and the more hydrophilic component in water is observed. This area of surface science requires increased attention because of the predominance of polymers as structural materials and as biomaterials. PMID- 11893062 TI - High-throughput single molecule screening of DNA and proteins. AB - We report a novel imaging technology for real time comprehensive analysis of molecular alterations in cells and tissues appropriate for automation and adaptation to high-throughput applications. With these techniques it should eventually be possible to perform simultaneous analysis of the entire contents of individual biological cells with a sensitivity and selectivity sufficient to determine the presence or absence of a single copy of a targeted analyte (e.g., DNA region, RNA region, protein), and to do so at a relatively low cost. The technology is suitable for DNA and RNA through sizing or through fluorescent hybridization probes, and for proteins and small molecules through fluorescence immunoassays. This combination of the lowest possible detection limit and the broadest applicability to biomolecules represents the final frontier in bioanalysis. The general scheme is based on novel concepts for single molecule detection (SMD) and characterization recently demonstrated in our laboratory. Since minimal manipulation is involved, it should be possible to screen large numbers of cells in a short time to facilitate practical applications. This opens up the possibility of finding single copies of DNA or proteins within single biological cells for disease markers without performing polymerase chain reaction or other biological amplification. PMID- 11893063 TI - Tryptophan synthase: a multienzyme complex with an intramolecular tunnel. AB - Tryptophan synthase is a classic enzyme that channels a metabolic intermediate, indole. The crystal structure of the tryptophan synthase alpha2beta2 complex from Salmonella typhimurium revealed for the first time the architecture of a multienzyme complex and the presence of an intramolecular tunnel. This remarkable hydrophobic tunnel provides a likely passageway for indole from the active site of the alpha subunit, where it is produced, to the active site of the beta subunit, where it reacts with L-serine to form L-tryptophan in a pyridoxal phosphate-dependent reaction. Rapid kinetic studies of the wild type enzyme and of channel-impaired mutant enzymes provide strong evidence for the proposed channeling mechanism. Structures of a series of enzyme-substrate intermediates at the alpha and beta active sites are elucidating enzyme mechanisms and dynamics. These structural results are providing a fascinating picture of loops opening and closing, of domain movements, and of conformational changes in the indole tunnel. Solution studies provide further evidence for ligand-induced conformational changes that send signals between the alpha and beta subunits. The combined results show that the switching of the enzyme between open and closed conformations couples the catalytic reactions at the alpha and beta active sites and prevents the escape of indole. PMID- 11893064 TI - Hydratases involved in nitrile conversion: screening, characterization and application. AB - The discovery of new enzymes with greater activity and specificity opens new, simple routes for synthetic processes, and consequently, new methods to solve environmental problems. A number of nitrile-related enzymes have been screened over the past few years for use in developing synthetic applications. Microbial nitrile hydratase (NHase) has great potential as a catalyst in organic chemical processing because the enzyme can convert nitriles to the corresponding higher value amides under mild conditions, and has now been applied to the industrial productions of acrylamide and nicotinamide. Particularly, the former production is the first successful example of a bioconversion process for the manufacture of a commodity chemical. The characterization of the enzyme at the molecular level has provided new insights into how the molecular structure determines the enzyme function, and how the regulatory system controls the expression of the enzyme genes to improve the enzyme and the NHase-dependent process. PMID- 11893065 TI - Bioorganic chemistry a la baguette: studies on molecular recognition in biological systems using rigid-rod molecules. AB - Initial studies using rigid-rod molecules or "baguettes" to address bioorganic topics of current scientific concern are reported. It is illustrated how transmembrane oligo(p-phenylene)s as representative model rods can be tuned to recognize lipid bilayer membranes either by their thickness or polarization. The construction of otherwise problematic hydrogen-bonded chains along transmembrane rods yields "proton wires," which act by a mechanism that is central in bioenergetics but poorly explored by means of synthetic models. Another example focuses on multivalent ligands assembling rigid-rod cell-surface receptors into transmembrane dynamic arene arrays. The potassium transport mediated by these ligand-receptor complexes provides experimental support for the potential biological importances of the controversial cation-pi mechanism. More complex supramolecular architecture is portrayed in the first artificial beta-barrels. It is shown how programmed assembly of toroidal rigid-rod supramolecules in detergent-free water permits control of diameter of the chemical nature of their interior. Reversed rigid-rod beta-barrels are assembled to function as self assembled ionophores, ion channel models, and transmembrane nanopores. The potential of future intratoroidal chemistry is exemplified by encapsulation and planarization of beta-carotene in water and the construction of transmembrane B DNA at the center of a second-sphere host-guest complex a al baguette. PMID- 11893066 TI - New approaches to developing lithium polymer batteries. AB - The electrochemical and physical-chemical properties of two families of lithium ion conducting membranes, i.e., the blends between high molecular weight poly(ethylene oxide) with a lithium salt commonly named "polymer electrolytes" and the gels of liquid solutions in a polymer matrix commonly named "gel electrolytes," are repoted and discussed. Particular attention is devoted to the newly developed approach of dispersing ceramic powders at the nanoscale particle dimension into the two types of membranes. This leads "nanocomposite" membranes having unique features, such as improved transport and interfacial properties in the case of the polymer electrolytes and enhanced liquid retention capability in the case of the gel electrolytes. Finally, the use of the gel electrolytes for the development of new-design, plastic-like, lithium-ion batteries is illustrated. PMID- 11893067 TI - Self and nonself recognition of chiral catalysts: the origin of nonlinear effects in the amino-alcohol catalyzed asymmetric addition of diorganozincs to aldehydes. AB - Asymmetric addition of dialkylzincs to aldehydes in the presence of (2S)-3-exo (dimethylamino)isoborneol [(S)-DAIB] exhibits various nonclassical phenomena. The enantiomeric excess (ee) of the alkylation product, obtained with partially resolved DAIB, is much higher than that of the chiral amino alcohol, while the rate decreases considerably as the ee of DAIB is lowered. The asymmetric amplification effects reflect the relative turnover numbers of two enantiomorphic catalytic cycles, where an essential feature is the reversible homochiral and heterochiral dimerization of the coexisting enantiomeric DAIB-based Zn catalysts. The interplay between the thermodynamics of the monomer/dimer equilibration and the kinetics of alkylation reaction strongly affect the overall profile of asymmetric catalysis. The self and nonself recognition of the chiral Zn catalysts is a general phenomenon when (S)-DAIB is mixed with its enantiomer, diastereomer, or even an achiral beta-amino alcohol. The degree of nonlinearity is highly affected not only by the structures and purity of catalysts but also by various reaction parameters. The salient features have been clarified on the basis of molecular weight measurements, NMR and X-ray crystallographic studies of organozinc complexes, and kinetic experiments, as well as computer-aided quantitative analysis. PMID- 11893068 TI - The role of radicals in enzymatic processes. AB - Research on the mechanism of action of coenzyme B12, adenosylcobalamin, as a graduate student introduced the author to the field of organic free radicals in enzymology. Twenty years later, related work on S-adenosylmethionine (SAM) as a "poor man's coenzyme B12" was initiated in a detailed analysis of the mechanism of action of lysine 2,3-aminomutase (LAM). The interconversion of L-lysine and L beta-lysine is catalyzed by LAM, which requires SAM, pyridoxal-5'-phosphate (PLP), and a [4Fe-4S] cluster as coenzymes. The mechanism of this reaction has been delineated as a radical isomerization, in which radical formation is initiated by the [4Fe-4S]-dependent cleavage of the SAM into methionine and the 5'-deoxyadenosyl radical. The mechanism of this process is discussed, together with the role of this radical in hydrogen abstraction from lysine to initiate the substrate radical isomerization. The chemistry underlying the functions of SAM, PLP, and [4Fe-4S] in the action of LAM is novel in all respects, except for the formation of a lysine-PLP aldimine at the active site. Of the four free radicals in the mechanism, three have been characterized by EPR spectroscopy. In the suicide inactivation of adenosylcobalamin-dependent dioldehydrase (DDH) by glycolaldehyde, the formation of cob(II)alamin and 5'-deoxyadenosine is accompanied by the conversion of glycolaldehyde to cis-ethanesemidione radical at the active site. The cis-ethanesemidione radical has been characterized by EPR spectroscopy. Its exceptional stability at the active site is the basis for the inactivation of DDH by glycolaldehyde. PMID- 11893069 TI - Weak interactions and molecular recognition in systems involving electron transfer proteins. AB - Electrostatic interactions and other weak interactions between amino acid side chains on protein surfaces play important roles in molecular recognition, and the mechanism of their intermolecular interactions has gained much interest. We established that charged peptides are useful for investigating the molecular recognition character of proteins and their molecular interaction induced structural changes. Positively charged lysine peptides competitively inhibited electron transfer from reduced cytochrome f (cyt f or cytochrome c (cyt c) to oxidized plastocyanin (PC), due to neutralization of the negatively charged site of PC by formation of PC-lysine peptide complexes. Lysine peptides also inhibited electron transfer from cyt c to cytochrome c peroxidase. Likewise, negatively charged aspartic acid peptides interacted with the positively charged sites of cytfand cyt c, and competitively inhibited electron transfer from reduced cytfor cyt c to oxidized PC and from [Fe(CN)6]4- to oxidized cyt c. Changes in the geometry and a shift to a higher redox potential of the active site Cu of PC on oligolysine binding were detected by spectroscopic and electrochemical measurements, owing to the absence of absorption in the visible region for lysine peptides. Structural and redox potential changes were also observed for cyt f and cyt c by interaction with aspartic acid peptides. PMID- 11893070 TI - The biosynthesis of acarbose and validamycin. AB - The studies reported here have established the biosynthetic origin of the mC7N units of acarbose and validamycin from sedo-heptulose 7-phosphate, and have identified 2-epi-5-epi-valiolone as the initial cyclization product. The deoxyhexose moiety of acarbose arises from glucose with deoxythymidyl-diphospho-4 keto-6-deoxy-D-glucose (dTDP-4-keto-6-deoxy-D-glucose) as a proximate intermediate. However, despite the identical origin of the aminocyclitol moieties in acarbose and validamycin A, the pathways of their formation seem to be substantially different. Validamycin A formation involves a number of discrete ketocyclitol intermediates, 5-epi-valiolone, valienone, and validone, whereas no free intermediates have been identified on the pathway from 2-epi-5-epi-valiolone to the pseudodisaccharide moiety of acarbose. The stage is now set for unraveling the mechanism or mechanisms by which the two components of the pseudodisaccharide moieties of acarbose and validamycin are uniquely coupled to each other via a nitrogen bridge. PMID- 11893071 TI - The sclerophytin A adventure. AB - The structural designation A originally made by Sharma and Alam to sclerophytin A was considered to be ambiguous and so notably strained relative to B that the latter was targeted for de novo synthesis (Scheme 1). Our two successful routes began with (5S)-(d-menthyloxy)-2(5H)-furanone and involved the application of cycloaddition, Claisen ring expansion, transannular oxymercuration, and 1,2 carbonyl transposition tactics to arrive at B. It was immediately apparent from polarity considerations and spectroscopic data that the antileukemic marine metabolite in question was in need of more deep-seated structural revision. Following close re-examination of an acquired authentic sample by advanced NMR techniques, the strong inference was made that sclerophytin A actually lacked a second oxygen bridge and was in reality the triol C. This conclusion was unequivocally confirmed by diverting an advanced intermediate generated earlier into a short sequence beginning with regiocontrolled dihydroxylation and terminating with configurational inversion at the secondary carbinol center. The status of other members of this series is also presented. PMID- 11893072 TI - Asymmetric autocatalysis and the origin of chiral homogeneity in organic compounds. AB - The discovery and development of asymmetric autocatalysis, in which the structures of the chiral catalyst and the chiral product are the same, are described. Chiral 5-pyrimidyl, 3-quinolyl, and 5-carbamoyl-3-pyridyl alkanols act as highly enantioselective asymmetric autocatalysts in the enantioselective addition of diisopropylzinc to the corresponding aldehydes, such as pyrimidine-5 carbaldehyde. 2-Alkynyl-5-pyrimidyl alkanol with an enantiomeric excess (ee) of >99.5% automultiplies practically perfectly as an asymmetric autocatalyst in a yield of >99% and >99.5% ee. Asymmetric autocatalysis with an amplification of ee has thus been realized. Consecutive asymmetric autocatalysis starting with chiral 2-alkynylpyrimidyl alkanol of only 0.6% ee amplifies its ee significantly, and yields itself as the product with >99.5% ee. The reaction of pyrimidine-5 carbaldehyde and diisopropylzinc in the presence of chiral initiators with low ee's, such as secondary alcohol, amine, carboxylic acid, mono-substituted [2.2]paracyclophane, and chiral primary alcohols due to deuterium substitution, regulates the absolute configuration of the resulting pyrimidyl alkanols, and the ee of the resulting pyrimidyl alkanol is much higher than that of the chiral initiator. Leucine and [6]helicene with very low ee's, which are known to be induced by circularly polarized light (CPL), also serve as chiral initiators to produce pyrimidyl alkanol with higher ee's. Overall, the process represents the first correlation between the chirality of CPL and an organic compound with very high ee. Chiral inorganic crystals, such as quartz and sodium chlorate, act as chiral inducers in the asymmetric autocatalysis of pyrimidyl alkanol. The process correlates for the first time ever the chirality of inorganic crystals with an organic compound with very high ee. PMID- 11893073 TI - Secondary metabolism in simulated microgravity. AB - We have studied microbial secondary metabolism in a simulated microgravity (SMG) environment provided by NASA rotating-wall bioreactors (RWBs). These reactors were designed to simulate some aspects of actual microgravity that occur in space. Growth and product formation were observed in SMG in all cases studied, i.e., Bacillus brevis produced gramicidin S (GS), Streptomyces clavuligerus made beta-lactam antibiotics, Streptomyces hygroscopicus produced rapamycin, and Escherichia coli produced microcin B17 (MccB17). Of these processes, only GS production was unaffected by SMG; production of the other three products was inhibited. This was determined by comparison with performance in an RWB positioned in a different mode to provide a normal gravity (NG) environment. Carbon source repression by glycerol of the GS process, as observed in shaken flasks, was not observed in the RWBs, whether operated in the SMG or NG mode. The same phenomenon occurred in the case of MccB17 production, with respect to glucose repression. Thus, the negative effects of carbon source on GS and beta lactam formation are presumably dependent on shear, turbulence, and/or vessel geometry, but not on gravity. Stimulatory effects of phosphate and the precursor L-lysine on beta-lactam antibiotic production, as observed in flasks, also occurred in SMG. An almost complete shift in the localization of produced MccB17 from cells to extracellular medium was observed when E. coli was grown in the RWB under SMG or NG. If a plastic bead was placed in the RWB, accumulation became cellular, as it is in shaken flasks, indicating that sheer stress favors a cellular location. In the case of rapamycin, the same type of shift was observed, but it was less dramatic, i.e., growth in the RWB under SMG shifted the distribution of produced rapamycin from 2/3 cellular:1/3 extracellular to 1/3 cellular:2/3 extracellular. Stress has been shown to induce or promote secondary metabolism in a number of other microbial systems. RWBs provide a low stress SMG environment, which, however, supports only poor production of MccB17, as compared to production in shaken flasks. We wondered whether the poor production in RWBs under SMG is due to the low level of stress, and whether increasing stress in the RWBs would raise the amount of MccB17 formed. We found that increasing shear stress by adding a single Teflon bead to the RWB improved MccB17 production. Although shear stress seems to have a marked positive effect on MccB17 production in SMG, addition of various concentrations of ethanol to RWBs (or to shaken flasks) failed to increase MccB17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the RWB were much more resistant to the growth- and production inhibitory effects of ethanol than cells growing in shaken flasks. With respect to S. hygroscopicus, addition of Teflon beads to the RWB reversed the inhibition of growth, but rapamycin production was still markedly inhibited, and the distribution did not revert back to a preferential cellular site. PMID- 11893074 TI - Immortalization of bovine umbilical vein endothelial cells: a model for the study of vascular endothelium. AB - Endothelial cells perform a large array of physiological functions that are influenced by their cellular heterogeneity in the different vascular beds. Vein endothelial cells isolated from the umbilical cords are commonly used to study vascular endothelium. Primary cultures of these cells, however, have low proliferative capacity and a limited life span. We have immortalized bovine umbilical vein endothelial cells (BUVEC) by transfection with an expression vector containing the human papillomavirus type 16 E6E7 oncogenes. Expression of E6E7 extended the life span of BUVEC from 40 to more than 1-20 cell replication cycles with no signs of senescence. Four immortalized clones were isolated and found to maintain endothelial cell properties, such as the uptake of acetylated low density lipoprotein, the expression of the von Willebrand protein, the binding of endothelial cell-specific lectins and proliferative responses to the specific endothelial cell mitogen, vascular endothelial growth factor. Moreover, clone BVE-E6E7-1, like its wild-type counterparts, expressed prolactin mRNA and decreased its proliferation in response to the anti-angiogenic 16-kDa fragment of prolactin. This clone showed little signs of genetic instability as revealed by centrosome and chromosome number analysis. Thus, immortalized E6E7 BUVEC cell lines retain endothelial cell characteristics and could facilitate studies to investigate the action of regulatory factors of vascular endothelium. Moreover, being the first non-human umbilical vein endothelial cell lines, their use should provide insights into the mechanisms governing species-related heterogeneity of endothelial cells. PMID- 11893075 TI - Pathogenic Mycobacterium avium remodels the phagosome membrane in macrophages within days after infection. AB - As part of their strategy for intracellular survival, mycobacteria prevent maturation of the phagosomes in which they reside inside macrophages. The molecular basis for this inhibition is only now beginning to emerge, by way of the molecular characterisation of the phagosome membrane when it encloses virulent mycobacteria. Our own work has shown that at 15 days after the phagocytic uptake of Mycobacterium avium by mouse bone marrow-derived macrophages, the phagosome membrane is depleted about 4-fold for cell surface derived membrane glycoconjugates, labelled by exogalactosylation, in comparison to the membrane of early endosomes with which it continues to interact. Here we asked whether this depletion occurred at early or late stages after infection. We found that only about half of the depletion had occurred at about 5 hours after the beginning of phagocytic uptake, with the remainder becoming established thereafter, with a half-time of about 2.5 days. Phagosomes became depleted in relation to early endosomes with which they continued to exchange membrane constituents. Early endosomes themselves became gradually depleted by about 30% during the 15-day post-infection period. In contrast, late endosomes/lysosomes remained unchanged, with a concentration of surface-derived glycoconjugates between that of early endosomes and of phagosomes at day 15 post infection. In view of the slowness of the post-infection change of phagosome membrane composition, we proposed that this change did not play a role in preventing maturation immediately after phagosome formation, but rather correlated with the process of maintaining the phagosomes in an immature state. PMID- 11893077 TI - Antisense CD11b integrin inhibits the development of a differentiated monocyte/macrophage phenotype in human leukemia cells. AB - Macrophage-like development of myeloid leukemia cells which can be induced by agents such as phorbol esters (TPA) is accompanied by integrin expression and cell adhesion. Thus, in differentiating myeloid leukemia cells CD11b is predominantly expressed which can associate with CD18 to form the functional heterodimeric integrin Mac-1. To elucidate the role of cell adhesion during macrophage-like differentiation, we transfected human U937 myeloid leukemia cells with a vector containing the CD11b gene in antisense orientation. Expression of the CD11b antisense gene in stably transfected U937 cells (as-CD11b cells) resulted in an attenuated response to TPA. As-CD11b cells demonstrated poor adhesion to solid substrate upon TPA treatment in contrast to U937 control cells. Constitutive expression of c-myc in as-CD11b transfectants was higher than in control cells and failed to be repressed by TPA treatment. Moreover, unlike control cells, antisense transfectants failed to induce expression of early response genes such as c-jun and the redox factor ref-1 upon TPA stimulation. Consequently, the induction of monocytic differentiation markers such as the activity of alpha-naphthyl acetate esterase, the capacity to reduce nitroblue tetrazolium and the expression of the vimentin gene was much lower in antisense transfectants than in control U937 cells. According to the failure to undergo a monocytic differentiation program, TPA treatment of as-CD11b cells resulted in a progressively increasing amount of apoptotic cells whereas the differentiated population of U937 control cells remained alive. Taken together, these data suggest that the integrin-mediated (particularly CD11b-mediated) adhesion of myeloid leukemia cells in the course of induced monocytic differentiation is crucial for cell attachment, development of a monocytic phenotype and subsequent survival. PMID- 11893076 TI - Molecular complexes that contain both c-Cbl and c-Src associate with Golgi membranes. AB - Cbl is an adaptor protein that is phosphorylated and recruited to several receptor and non-receptor tyrosine kinases upon their activation. After binding to the activated receptor, Cbl plays a key role as a kinase inhibitor and as an E3 ubiquitin ligase, thereby contributing to receptor down-regulation and internalization. In addition, Cbl translocates to intracellular vesicular compartments following receptor activation. We report here that Cbl also associates with Golgi membranes. Confocal immunofluorescence staining of Cbl in a variety of unstimulated cells, including CHO cells, revealed a prominent perinuclear colocalization of Cbl and a Golgi marker. Both the prominent Cbl staining and the Golgi marker were dispersed by brefeldin A. Subcellular fractionation of CHO cells demonstrated that about 10% of Cbl is stably associated with membranes, and that Golgi-enriched membrane fractions produced by isopycnic density centrifugation and free-flow electrophoresis are also enriched in Cbl, relative to other membrane fractions. The membrane-bound Cbl was hyperphosphorylated and it co-immunoprecipitated with endogenous Src. By immunofluorescence, some Src colocalized with Cbl and Golgi markers, and Src, like Cbl, was present in the Golgi-enriched fraction prepared by sequential density centrifugation and free-flow electrophoresis. Transfection of an activated form of Src, but not wild-type Src, increased the amount of Src that co immunoprecipitated with Cbl, and increased the intensity of Cbl staining on the Golgi. This result, together with the increased tyrosine phosphorylation of the membrane-associated Cbl, suggests that Golgi-associated Cbl could be part of a molecular complex that contains activated Src. The localization and interaction of Src and Cbl at the Golgi and the regulation of the interaction of Cbl with Golgi membrane suggest that this complex may contribute to the regulation of Golgi function. PMID- 11893078 TI - The role of fetal and adult hepatocyte extracellular matrix in the regulation of tissue-specific gene expression in fetal and adult hepatocytes. AB - We explored the effect of extracellular matrix (ECM) produced by fetal and adult hepatocytes on tissue-specific gene expression and proliferation of fetal and adult hepatocytes. Adult hepatocytes ECM strongly induced expression of both albumin and HNF-4 in adult hepatocytes. In contrast, fibroblast ECM reduced the expression of mRNAs for albumin and alpha-fetoprotein in fetal hepatocytes. Adult hepatocytes ECM also increased the activity of liver-specific enzymes of adult hepatocytes (DPP IV and glucose-6-phosphatase) in both fetal and adult hepatocytes, while fetal hepatocyte-derived ECM increased activity of the fetal hepatocyte enzyme GGT in fetal hepatocytes. Fibroblast ECM was inhibitory for the activity of all enzymes assayed. Removal of heparin chains from the various matrices by pretreatment of the ECM with heparinase resulted in reduction of glucose-6-phosphatase and DPP IV in adult hepatocytes. Removal of chondroitin sulfate chains from fetal hepatocyte-derived ECM resulted in loss of induction of GGT in the fetal cells. Fetal hepatocytes proliferated best on adult hepatocyte derived ECM. Adult hepatocytes showed only modest proliferation on both fetal and adult hepatocytes ECM and their growth was inhibited by fibroblast ECM. In conclusion, adult hepatocyte ECM better supports the expression of adult genes, whereas fetal hepatocyte ECM induced expression of fetal genes. Fibroblast derived-ECM was inhibitory for both proliferation and tissue-specific gene expression in fetal and adult hepatocytes. The data support a role for heparan sulfate being the active element in adult ECM, and chondroitin sulfate being the active element in fetal ECM. PMID- 11893079 TI - DNA catenations that link sister chromatids until the onset of anaphase are maintained by a checkpoint mechanism. AB - Treatment of Allium cepa meristematic cells in metaphase with the topoisomerase II inhibitor ICRF-193, results in bridging of the sister chromatids at anaphase. Separation of the sisters in experimentally generated acentric chromosomal fragments was also inhibited by ICRF-193, indicating that some non-centromeric catenations also persist in metaphase chromosomes. Thus, catenations must be resolved by DNA topoisomerase II at the metaphase-to-anaphase transition to allow segregation of sisters. A passive mechanism could maintain catenations holding sisters until the onset of anaphase. At this point the opposite tension exerted on sister chromatids could render the decatenation reaction physically more favorable than catenation. But this possibility was dismissed as acentric chromosome fragments were able to separate their sister chromatids at anaphase. A timing mechanism (a common trigger for two processes taking different times to be completed) could passively couple the resolution of the last remaining catenations to the moment of anaphase onset. This possibility was also discarded as cells arrested in metaphase with microtubule-destabilising drugs still displayed anaphase bridges when released in the presence of ICRF-193. It is possible that a checkpoint mechanism prevents the release of the last catenations linking sisters until the onset of anaphase. To test whether cells are competent to fully resolve catenations before anaphase onset, we generated multinucleate plant cells. In this system, the nuclei within a single multinucleate cell displayed differences in chromosome condensation at metaphase, but initiated anaphase synchronously. When multinucleates were treated with ICRF-193 at the metaphase-toanaphase transition, tangled and untangled anaphases were observed within the same cell. This can only occur if cells are competent to disentangle sister chromatids before the onset of anaphase, but are prevented from doing so by a checkpoint mechanism. PMID- 11893080 TI - Joint participation of mitochondria and sarcoplasmic reticulum in the formation of tubular aggregates in gastrocnemius muscle of CK-/- mice. AB - Tubular aggregates are specific subcellular structures that appear in skeletal muscle fibres under different pathological conditions. The origin of the tubular aggregates is generally ascribed to proliferating membranes of sarcoplasmic reticulum. There are, however, histochemical indications for the presence of mitochondrial enzymes in tubular aggregates suggesting contribution of mitochondria to the genesis of tubular aggregates. In this study we used an immunocytochemical detection technique to assess participation of mitochondria and of sarcoplasmic reticulum in derivation of tubular aggregates. The fast skeletal muscle fibres (m. gastrocnemius) of mice bearing the double invalidation for both the mitochondrial and the cytosolic isoforms of creatine kinase (CK), an enzyme involved in energetics of muscle cells, were employed as a model muscle with tubular aggregates (Steeghs et al., Cell 89, 93-103, 1997). Immunogold labelling of the bc1 complex, a specific integral protein of the inner mitochondrial membrane, provided strong signals in both the mitochondria and tubular aggregates but not in other ultrastructural components of muscle fibres. A similar strong immunogold signal was obtained when labelling for SERCA1, a specific enzyme of the sarcoplasmic reticulum membrane, in regions of typical occurrence of the sarcoplasmic reticulum and in tubular aggregates. In double labelling experiments, we found simultaneous labelling of tubular aggregates with both the bc1 and SERCA1 antibodies. It is concluded, that in CK-/- mouse both the inner mitochondrial membrane and the membrane of the sarcoplasmic reticulum participate in the formation of tubular aggregates. PMID- 11893081 TI - Immunocytochemical localization of allergenic proteins from mature to activated Zygophyllum fabago L. (Zygophyllaceae) pollen grains. AB - Zygophyllum fabago L. (Zygophyllaceae) can be found in the Middle East, in North Africa and in the arid zones of the Mediterranean region. It easily establishes itself in new regions, and is considered an invasive plant. They undergo ambophilous pollination, as there is a relationship between this type of pollination and its allergenic incidence. A combination of transmission electron microscopy with immunocytochemical methods was used to localize allergenic proteins during hydration and activation processes. Germination was induced in vitro for 1,2,4,6, and 30 min. The activated proteins reacting with antibodies present in human sera from allergenic patients are found in the cytoplasm, intine, exine and exudates from the pollen grains. The activation time plays an important role on the labelling intensity. Labelling of allergenic proteins was abundant at 1 and 2 min of activation, and decreased at 4 and 6 min. The rapid activation and release of the allergenic proteins appears to be the main cause of allergenic activity of Z. fabago pollen grains. PMID- 11893082 TI - Conservation of the gene structure and membrane-targeting signals of germ cell specific lamin LIII in amphibians and fish. AB - Targeting of nuclear lamins to the inner nuclear membrane requires CaaX motif dependent posttranslational isoprenylation and carboxyl methylation. We previously have shown that two variants of lamin LIII (i.e., LIII and LIIIb) in amphibian oocytes are generated by alternative splicing and differ greatly in their membrane association. An extra cysteine residue (as a potential palmitoylation site) and a basic cluster in conjunction with the CaaX motif function as secondary targeting signals responsible for stable membrane association of lamin LIIIb. cDNA sequencing and genomic analysis of the zebrafish Danio rerio lamin LIII uncovers a remarkable conservation of the genomic organization and of the two secondary membrane-targeting signals in amphibians and fish. The expression pattern of lamin LIII genes is also conserved between amphibians and fish. Danio lamin LIII is expressed in diplotene oocytes. It is absent from male germ cells but is expressed in Sertoli cells of the testis. In addition, we provide sequence information of the entire coding sequence of zebrafish lamin A, which allows comparison of all major lamins from representatives of the four classes of vertebrates. PMID- 11893083 TI - Distinct cell type-specific expression of scaffolding proteins EBP50 and E3KARP: EBP50 is generally expressed with ezrin in specific epithelia, whereas E3KARP is not. AB - The ezrin/radixin/moesin (ERM) proteins are regulated microfilament membrane linking proteins. Previous tissue localization studies have revealed that the three related proteins show distinct tissue distributions, with ezrin being found predominantly in polarized epithelial cells, whereas moesin is enriched in endothelial cells and lymphocytes. EBP50 and E3KARP are two related scaffolding proteins that bind to the activated form of ERM proteins in vitro, and through their PDZ domains to the cytoplasmic domains of specific membrane proteins, including the Na+/H+ exchanger isoform (NHE3) present in kidney proximal tubules and the beta2-adrenergic receptor. Using specific antibodies to EBP50 and E3KARP for localization in murine tissues, we find that the cellular distribution of EBP50 and E3KARP is mutually exclusive. Epithelial cells expressing ezrin generally co-express EBP50, such as intestinal epithelial cells, gastric parietal cells, the epithelial cells of the kidney proximal tubule, the terminal bronchiole of the lung, and in mesothelia. This correlation is not absolute as cells of the mucous epithelium of the stomach and in the renal corpuscle, express ezrin but no detectable EBP50, whereas the bile canaliculi of hepatocytes express EBP50 and not ezrin. E3KARP has a restricted tissue distribution with the highest expression being found in lung. It is largely colocalized with moesin and radixin, especially in the alveoli of the lung, as well as being highly enriched in the renal corpuscle. These results document a preference for co-expression of EBP50, but not E3KARP, with ezrin in polarized epithelia. These results place constraints on the physiological roles that can be proposed for these scaffolding molecules. PMID- 11893084 TI - DNA-containing extracellular 50-nm particles in the ileal Peyer's patch of sheep. AB - Follicles of the ileal Peyer's patch are sites of B cell proliferation and of diversification of the primary immunoglobulin repertoire in ruminants. We demonstrate here that 50-nm carbonic anhydrase-reactive particles released in the intercellular space in the follicle-associated epithelium of the ileal Peyer's patch of lambs contain DNA protected with a detergent-resistant membrane. We named these particles DiCAPs (DNA in carbonic anhydrase particles). DiCAPs can be purified from a suspension collected from ileal Peyer's patch follicles by sedimentation in a sucrose gradient. The DiCAP membrane is resistant to several ionic and non-ionic detergents alone, but can be disrupted by a combination of Triton X-100 and proteinase K. Differential nuclease treatment of purified DiCAPs indicates that they contain DNA. Digestion of DiCAP DNA with six-base pair restriction enzymes produces smears, suggesting that individual DiCAPs contain unique sequences. Nonetheless, the size of DiCAP DNA is smaller (approximately 16 kb) than that of lamb genomic DNA. Polymerase chain reaction and sequence analysis of DiCAP DNA reveals the presence of light and heavy chain variable genes as well as housekeeping genes. The data demonstrate the presence of DNA in these extracellular particles, and suggest a role of DiCAPs in transfer of DNA between cells within the ileal Peyer's patch. This raises the possibility of a novel form of communication between cells mediated by nucleic acids. PMID- 11893085 TI - Interaction of gamma-COP with a transport motif in the D1 receptor C-terminus. AB - Truncations at the carboxyl termini of G protein-coupled receptors result in defective receptor biogenesis and comprise a number of inherited disorders. In order to evaluate the structural role of the C-terminus in G protein-coupled receptor biogenesis, we generated a series of deletion and substitution mutations in the dopamine D1 receptor and visualized receptor subcellular localization by fusion to a green fluorescent protein. Alanine substitutions of several hydrophobic residues within the proximal C-terminus resulted in receptor transport arrest in the ER. Agonist binding and coupling to adenylyl cyclase was also abolished. In contrast, substitutions conserving C-terminal hydrophobicity produced normal cell surface receptor expression, binding, and stimulatory function. A mechanism for the role of the C-terminus in D1 receptor transport was investigated by searching for candidate protein interactions. The D1 receptor was found to co-precipitate and associate in vitro directly with the gamma-subunit of the COPI coatomer complex. In vitro pull-down assays confirmed that only the D1 C terminus is required for COPI association, and that identical mutations causing disruption of receptor transport to the cell surface also disrupted binding to COPI. Furthermore, conservative mutations in the D1 C-terminus restored COPI association just as they restored cell surface transport. These results suggest that association between the coatomer complex and hydrophobic residues within the proximal C-terminus of the D1 receptor may serve an important role in receptor transport. PMID- 11893087 TI - The interactive effects of infant activity level and fear on growth trajectories of early childhood behavior problems. AB - The current study examined the interactive effects of infant activity level and fear on growth trajectories of behavior problems in early childhood (age 4 to 8 years) using maternal ratings. The sample was drawn from the National Longitudinal Survey of Youth (NLSY) and included children who were between 1 and 11 months in 1986. Findings suggested that boys characterized by high activity level and low levels of fear in infancy escalated in both externalizing and internalizing symptoms. Also, boys characterized by high fear and low activity level increased in internalizing symptoms and these effects seemed to be specific to depression rather than anxiety. Temperament did not predict escalation in externalizing symptomatology for girls, but low levels of fear predicted increases in internalizing symptoms. There was also evidence for a decline in depression specific symptoms for girls characterized by high fear and low activity in infancy. These findings suggest the importance of examining interactive models of temperament risk and considering gender specific pathways to behavior problems. PMID- 11893086 TI - The calcium-binding protein p54/NEFA is a novel luminal resident of medial Golgi cisternae that traffics independently of mannosidase II. AB - A new Golgi resident, p54, has been demonstrated in several eukaryotic species and in multiple organs. Based on Triton X-114 partition, carbonate extraction and trypsin protection assays, p54 behaved as an extrinsic membrane protein, facing the luminal compartment. p54 was purified by two-dimensional electrophoresis and identified by matrix-assisted laser desorption ionization/time-of-flight (MALDI TOF) mass spectrometry as NEFA, a calcium-binding protein (Barnikol-Watanabe et al., 1994, Biol. Chem. Hoppe Seyler, 375, 497-512). By immunofluorescence, p54/NEFA essentially colocalized with the medial Golgi marker mannosidase II, and did not overlap with the cis-Golgi marker p58, nor with the trans-Golgi network (TGN) marker TGN38. By immuno-electron microscopy, p54/NEFA localized in the medial cisternae and in Golgi-associated vesicles. p54/NEFA remained associated with mannosidase II despite Golgi disruption by nocodazole, caffeine, or, to some extent, potassium depletion (a new procedure to induce Golgi disassembly), but the two markers rapidly dissociated upon brefeldin A treatment and at metaphase, and reassociated upon drug removal and at the end of anaphase. Since p54/NEFA is a peripheral luminal membrane constituent, its distinct trafficking from the transmembrane marker mannosidase II suggests a novel Golgi retention mechanism, by strong association of this soluble protein with another integral transmembrane resident. PMID- 11893088 TI - Response decision processes and externalizing behavior problems in adolescents. AB - Externalizing behavior problems of 124 adolescents were assessed across Grades 7 11. In Grade 9, participants were also assessed across social-cognitive domains after imagining themselves as the object of provocations portrayed in six videotaped vignettes. Participants responded to vignette-based questions representing multiple processes of the response decision step of social information processing. Phase 1 of our investigation supported a two-factor model of the response evaluation process of response decision (response valuation and outcome expectancy). Phase 2 showed significant relations between the set of these response decision processes, as well as response selection, measured in Grade 9 and (a) externalizing behavior in Grade 9 and (b) externalizing behavior in Grades 10-11, even after controlling externalizing behavior in Grades 7-8. These findings suggest that on-line behavioral judgments about aggression play a crucial role in the maintenance and growth of aggressive response tendencies in adolescence. PMID- 11893089 TI - Prediction of peer-rated adult hostility from autonomy struggles in adolescent family interactions. AB - Observed parent-adolescent autonomy struggles were assessed as potential predictors of the development of peer-rated hostility over a decade later in young adulthood in both normal and previously psychiatrically hospitalized groups of adolescents. Longitudinal, multireporter data were obtained by coding family interactions involving 83 adolescents and their parents at age 16 years and then obtaining ratings by close friends of adolescents' hostility at age 25 years. Fathers' behavior undermining adolescents' autonomy in interactions at age 16 years were predictive of adolescents-as-young-adults' hostility, as rated by close friends at age 25 years. These predictions contributed additional variance to understanding young adult hostility even after accounting for concurrent levels of adolescent hostility at age 16 years and paternal hostility at this age, each of which also significantly contributed to predicting future hostility. Results are discussed as highlighting a pathway by which difficulties attaining autonomy in adolescence may presage the development of long-term difficulties in social functioning. PMID- 11893090 TI - Parent alcoholism and the leaving home transition. AB - Although they have received little empirical attention, departures from the parental home play a significant role in demarcating the transition from adolescence to young adulthood. The current study examined the extent to which various features of young adults' experiences of leaving home differed for children of alcoholic (COAs) versus nonalcoholic parents, what adolescent precursors might account for noted differences and what indicators of young adult adjustment are related to the leaving home experience. A total of 227 young adults drawn from a high-risk, community sample of COAs and matched controls were interviewed at ages 18-23 years regarding their prior leaving home experiences. COAs showed greater difficulties in negotiating this transition, fewer positive feelings about the transition, and different reasons for leaving home as compared to participants without an alcoholic parent. Moreover, adolescent risk behaviors, family conflict, and family disorganization (assessed prior to this transition) each partly accounted for COAs' risk for difficulty in the leaving home transition. Although certain aspects of the leaving home transition were uniquely related to young adult adjustment, future research is still needed to more comprehensively understand the implications for young adult development associated with such individual differences in the leaving home transition. PMID- 11893091 TI - Male and female offending trajectories. AB - This paper uses a latent class modeling approach to examine gender related variations in offending trajectories from adolescence to young adulthood. This approach is applied to data gathered over the course of a longitudinal study of 896 New Zealand children studied from birth to age 21 years. The analysis identified five trajectory groups: a group of low-risk offenders, three groups of adolescent-limited offenders who varied in the timing of the onset of offending (early, intermediate, and late onset), and a group of chronic offenders. Identical offending trajectories applied for males and females. However, probabilities of trajectory group membership varied with gender, with females being more likely to exhibit low-risk or early onset adolescent-limited offending and males later onset and chronic offending. Examination of social, family, and individual factors associated with these trajectories suggested the presence of a series of common etiological factors relating to family functioning and early adjustment that discriminated between trajectory groups. These risk factors appeared to operate in a similar fashion for both males and females. Implications of these findings for trajectory theories of offending are discussed. PMID- 11893092 TI - Males on the life-course-persistent and adolescence-limited antisocial pathways: follow-up at age 26 years. AB - This article reports a comparison on outcomes of 26-year-old males who were defined several years ago in the Dunedin longitudinal study as exhibiting childhood-onset versus adolescent-onset antisocial behavior and who were indistinguishable on delinquent offending in adolescence. Previous studies of these groups in childhood and adolescence showed that childhood-onset delinquents had inadequate parenting, neurocognitive problems, undercontrolled temperament, severe hyperactivity, psychopathic personality traits, and violent behavior. Adolescent-onset delinquents were not distinguished by these features. Here followed to age 26 years, the childhood-onset delinquents were the most elevated on psychopathic personality traits, mental-health problems, substance dependence, numbers of children, financial problems, work problems, and drug-related and violent crime, including violence against women and children. The adolescent onset delinquents at 26 years were less extreme but elevated on impulsive personality traits, mental-health problems, substance dependence, financial problems, and property offenses. A third group of men who had been aggressive as children but not very delinquent as adolescents emerged as low-level chronic offenders who were anxious, depressed, socially isolated, and had financial and work problems. These findings support the theory of life-course-persistent and adolescence-limited antisocial behavior but also extend it. Findings recommend intervention with all aggressive children and with all delinquent adolescents, to prevent a variety of maladjustments in adult life. PMID- 11893093 TI - Spatial but not verbal cognitive deficits at age 3 years in persistently antisocial individuals. AB - Previous studies have repeatedly shown verbal intelligence deficits in adolescent antisocial individuals, but it is not known whether these deficits are in place prior to kindergarten or, alternatively, whether they are acquired throughout childhood. This study assesses whether cognitive deficits occur as early as age 3 years and whether they are specific to persistently antisocial individuals. Verbal and spatial abilities were assessed at ages 3 and 11 years in 330 male and female children, while antisocial behavior was assessed at ages 8 and 17 years. Persistently antisocial individuals (N = 47) had spatial deficits in the absence of verbal deficits at age 3 years compared to comparisons (N = 133), and also spatial and verbal deficits at age 11 years. Age 3 spatial deficits were independent of social adversity, early hyperactivity, poor test motivation, poor test comprehension, and social discomfort during testing, and they were found in females as well as males. Findings suggest that early spatial deficits contribute to persistent antisocial behavior whereas verbal deficits are developmentally acquired. An early-starter model is proposed whereby early spatial impairments interfere with early bonding and attachment, reflect disrupted right hemisphere affect regulation and expression, and predispose to later persistent antisocial behavior. PMID- 11893095 TI - A longitudinal analysis of patterns of adjustment following peer victimization. AB - This study examined the effects of being victimized by peers on children's behavioral, social, emotional, and academic functioning. We assessed an ethnically diverse sample of 2,064 first, second, and fourth graders and followed them over 2 years, locating 1,469 of the participants at the follow-up. Correlation and partial correlation analyses revealed that prior victimization predicted externalizing, internalizing, and social problems 2 years later for the sample as a whole. However, not all victimized children experienced the same types of outcomes; instead, there was heterogeneity in children's responses to victimization. Using cluster analysis, we identified eight outcome patterns that represented different patterns of functioning. These were labeled as externalizing, internalizing, symptomatic, popular, disliked, absent, low achieving, and high achieving. Discriminant function analyses revealed that the symptomatic, externalizing, and disliked patterns were systematically related to victimization. Moreover, significant gender and age differences in the severity of effects were obtained. The discussion highlights the complexity of victimization effects. PMID- 11893094 TI - Preschool children with disruptive behavior: three-year outcome as a function of adaptive disability. AB - A significant discrepancy between intelligence and daily adaptive functioning, or adaptive disability (AD), has been previously found to be a associated with significant psychological morbidity in preschool children with disruptive behavior (DB). The utility of AD as a predictor of later developmental risks was examined in a 3-year longitudinal study of normal (N = 43) and DB preschool children. The DB children were grouped into those with AD (DB+AD; N = 28) and those without AD (DB-only; N = 98). All children were followed with annual evaluations to the end of second grade. Both DB groups demonstrated substantial and pervasive psychological and educational morbidity at 3-year follow-up. In comparison to DB-only children, DB+AD children had more symptoms of attention deficit hyperactivity disorder (ADHD) and conduct disorder (CD), more severe and pervasive behavior problems at home, more parent-rated externalizing and internalizing, and lower academic competence and more behavioral problems at school. Parents of DB+AD children also reported greater parenting stress than did parents in the other groups. A significant contribution of AD to adverse outcomes in the DB group remained on some measures even after controlling for initial severity of DB. AD also contributed significantly to CD symptoms at follow-up after controlling for initial DB severity and initial CD symptoms. The results corroborate and extend earlier findings of the utility of AD as a risk indicator above severity of DB alone. They also imply that AD in the context of normal intellectual development may arise from both the deficient self-regulation associated with ADHD and from disrupted parenting. with exposure to kindergarten moderating these adverse effects. PMID- 11893096 TI - Peer to peer sexual harassment in early adolescence: a developmental perspective. AB - The goal of this study was to examine sexual harassment in early adolescence. Available data indicate that peer to peer sexual harassment is prevalent in high school and is associated with psychosocial problems for both victims and perpetrators. For the present study, we adopted a developmental contextual model to examine the possibility that this behavior develops during the late elementary and middle school years and is linked to the biological and social changes that occur at this time. Youths from Grades 6-8 (N = 1,213) enrolled in seven elementary and middle schools in a large south-central Canadian city were asked to report on their sexual harassment behaviors with same- and cross-gender peers; their pubertal development, and the gender composition of their peer network. The results revealed that cross-gender harassment was distinct from same-gender harassment, increased in frequency from Grade 6 to Grade 8, and was linked to pubertal maturation and participation in mixed-gender peer groups. The implications of a developmental contextual model for understanding the emergence of this problematic behavior in adolescence are discussed. PMID- 11893097 TI - Presidential address. The best of the Midwest. PMID- 11893098 TI - Senior surgical residents can accurately interpret trauma radiographs. AB - Surgical residents routinely interpret radiographic studies during the evaluation of trauma patients, which directs further evaluation and invasive procedures. Official interpretations--"post-reading"--of radiographs by radiologists may be delayed by hours or even days. Trauma surgeons frequently act on their impressions before "official" readings are available. It has been demonstrated that surgical residents can accurately perform and interpret trauma ultrasound examinations. The purpose of this study was to evaluate the ability of senior surgery residents to interpret basic trauma radiographs. Interpretations of trauma radiographs (cervical spine, chest, pelvis, and CT of the brain) were recorded prospectively by the senior surgery resident present during trauma evaluations. These interpretations were compared with the findings of the radiologist as obtained from the official radiology report. Differing results were divided into clinically significant and clinically nonsignificant findings using defined criteria. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were determined. Interpretations of trauma radiographs by senior residents achieved an accuracy of 100 per cent for cervical spine radiographs, 95.9 per cent for chest radiographs, 98.0 per cent for pelvis radiographs, and 97.9 per cent for CT of the head. In aggregate senior residents interpreted trauma radiographs with 97.9 per cent accuracy. Differences that were considered clinically significant according to preset criteria occurred in 2.1 per cent of observations. We conclude that senior general surgical residents can accurately interpret trauma radiology, including CT of the brain. These results suggest that institutional policies for post-reading of trauma radiology should be reassessed. PMID- 11893099 TI - Nonoperative management of blunt splenic injuries: factors influencing success in age >55 years. AB - Historically poor success rates of nonoperative management of splenic injuries in elderly patients have led to recommendations for operative intervention in patients more than 55 years of age. Recent studies are in opposition to earlier recommendations revealing equal success rates of nonoperative management of splenic injuries in all age groups. A retrospective chart review was performed to assess factors related to the successful management of splenic injuries in patients over 55 years of age at a Level I trauma center. Thirty-seven patients over 55 presented with blunt splenic injuries during the 5-year study period. Thirteen patients were taken immediately to the operating room on the basis of clinical findings and/or abdomen/pelvis CT results. Nonoperative management was attempted in 24 patients on the basis of CT findings. Nonoperative management was successful in 15 patients (62.5%) and failed in eight patients (33.3%). Patients who failed nonoperative management had significantly higher American Association for the Surgery of Trauma splenic injury grade and associated pelvic free fluid. There were no deaths related to complications from failed nonoperative management. We conclude that nonoperative management of blunt splenic injuries in patients over 55 may be attempted. Patients with higher-grade injuries and pelvic free fluid are at greater risk for failure. Patients with these two findings must be monitored closely. The physicians caring for elderly patients with high-grade splenic injuries and free fluid in the pelvis must use clinical judgment regarding the need and timing of operative management. PMID- 11893100 TI - Factors affecting the outcome of patients with splenic trauma. AB - This is a report of 546 consecutive patients with penetrating and blunt splenic trauma seen over a 17 1/2-year period (1980-1997). The etiology of the splenic injuries and the associated mortality rates were: blunt injuries 45 of 298 (15%), gunshot wounds 48 of 199 (24%), and stab wounds four of 49 (8%). The overall mortality rate was 97 of 546 (18%). The most significant risk factors for death were all associated with major blood loss: transfusion requirements > or = 6 units of blood, low initial operating room blood pressure, associated abdominal vascular injuries, and performance of a thoracotomy. The two most important organs injured in conjunction with the spleen that were significant predictors of postoperative infectious complications were colon and pancreas. The need for splenectomy was most significantly correlated with higher grades of splenic injury especially grades IV and V. The evolution in management of blunt splenic trauma has led to a significant improvement in splenic preservation and avoidance of laparotomy for many patients. Operative splenic salvage is reduced in patients subjected to laparotomy who are candidates for nonoperative treatment. Improved results with splenic injury should be obtained by rapid control of bleeding. This may require more liberal criterial in selecting patients with splenic trauma for early operative treatment. PMID- 11893101 TI - Burns with multiple trauma. AB - The purpose of this study was to determine the incidence, mechanisms, and outcomes of management in patients with multisystem trauma and associated burn injury. A retrospective review was performed of patients admitted with combined burns and trauma from 1990 through 1999. Mechanism of injury, extent of burns, associated injuries, Injury Severity Score (ISS), and patient outcomes were identified. There were 2,845 burn and 19,418 trauma admissions. Fifty-six patients (2.0% and 0.29% respectively) had combined burns and trauma. Mean ISS was 21.7 and average percentage total body surface area was 16.2. Associated injuries included fractures in 32, complex soft-tissue injury in 20, head injury in 11, and abdominal trauma in seven. Mechanism of injury was industrial in 19, motor vehicle accident in 16, house fire in 13, high voltage in six, and other in three. Skin grafting was required in 33 of 56 patients (59%). Six of 56 patients died. Mean ISS was 19.0 in survivors compared with 46.2 in nonsurvivors. The combination of burns with multiple system trauma is uncommon. Fractures are the most frequent associated injury, and the majority of patients will require skin grafting in their burn treatment. Outcomes with appropriate management are favorable and are primarily dependent on the degree of associated trauma. PMID- 11893102 TI - The clinical presentation and operative management of nodular and diffuse substernal thyroid disease. AB - Patients with substernal thyroid disease, defined by the presence of enlarged thyroid tissue below the plane of the thoracic inlet, were identified from a prospective database maintained for patients who have undergone thyroidectomy at our institution since 1990. Substernal thyroid disease was present in 116 (30%) of 381 patients, anterior mediastinal in 109 (94%), and posterior mediastinal in seven (6%). Indications for surgery included compressive symptoms in 75 (65%) patients, an abnormal fine-needle biopsy in 45 (39%), progressive thyroid enlargement in 41 (35%), thyrotoxicosis in 11 (10%), and superior vena cava syndrome in two (1.7%). A median sternotomy and thoracotomy were performed in one patient each for a primary intrathoracic goiter. In all other patients thyroidectomy was accomplished through a cervical incision. Parathyroid autotransplantation was performed in 41 (37%) patients with retrosternal disease compared with 57 (22%) with disease confined to the neck (P < 0.01). Twenty-five patients (22%) had malignancy; four of these had unresectable disease. Postoperative complications included transient hypocalcemia (n = 46), transient hoarseness (n = 7), recurrent laryngeal nerve injury (n = 1), and wound infection (n = 1). One patient died from aspiration pneumonia. In summary, substernal thyroid disease is typically present in the anterior mediastinum and with rare exceptions can be resected through a cervical incision. Parathyroid devascularization is more common with resection of a substernal goiter and autotransplantation can prevent permanent hypoparathyroidism. PMID- 11893103 TI - Primary hyperaldosteronism in the era of laparoscopic adrenalectomy. AB - Laparoscopic adrenalectomy has been recommended as the standard method for removing an aldosteronoma. To assess our surgical experience with primary hyperaldosteronism in the era of laparoscopic adrenalectomy a 6-year retrospective review of 30 consecutive patients was done. The 20 men and 10 women ranged in age from 35 to 78 with a mean of 51.2 years. All patients were hypertensive and hypokalemic with a mean serum potassium of 2.9 +/- 0.32 (standard deviation) mmol/L. Serum aldosterone was elevated in 28 of 30 (94%) patients and normal in the remaining two. Serum renin was suppressed in all patients. CT correctly localized the tumor in all 30 patients. Twenty-eight patients had histologically documented adenomas and two had associated cortical hyperplasia on pathology. Mean adenoma size was 2.0 +/- 1.12 cm. Twenty-four patients underwent left laparoscopic adrenalectomies, whereas right laparoscopic adrenalectomies were performed in five. One was converted to an open left adrenalectomy. Mean operative time was 183 minutes. The mean hospital stay for laparoscopic adrenalectomy was 2.2 days. The patients were followed from one to 63 months (mean 26.1 months). Twenty-nine of 30 (95%) patients were rendered normokalemic. The remaining patient takes a potassium-wasting diuretic. Persistent hypertension was present in 10 of 30 (33%) patients. Blood pressure in nine of 10 patients was controlled with less medical therapy. The other patient's blood pressure remained difficult to control despite multiple medications. Duration of hypertension before surgery was a significant risk factor for persistent hypertension (P < 0.05). Gender (P > 0.05) and age (P > 0.05) at the time of surgery were not statistically significant predicators for persistent hypertension. There were two reported trocar site hernias. We conclude that primary hyperaldosteronism due to aldosterone-producing tumors can be diagnosed and accurately localized with preoperative measurements of serum aldosterone, renin, and CT scanning. Laparoscopic adrenalectomy is safe and effective for the treatment of primary hyperaldosteronism with minimal associated morbidity and a short hospital stay. Hypokalemia may be cured by surgical treatment, although persistent hypertension still occurs. Duration of hypertension before surgery is a risk factor for persistent hypertension whereas age and sex are not. PMID- 11893104 TI - Prognostic factors associated with resectable carcinoma of the esophagus. AB - A retrospective review of esophagectomy for esophageal carcinoma between 1982 and 1999 was performed. Two hundred twenty-two patients (mean age 61.7 years) underwent esophagectomy: 128 transhiatal, 74 Ivor Lewis, and 20 abdominal. Most tumors were adenocarcinoma (65%); the majority were in the lower third or cardia (78%). Excluding operative mortality the one-, 3-, and 5-year survival rates were 67, 39, and 31 per cent (median survival, 16.3 months) respectively. The hospital mortality rate was 6.8 per cent. Through univariate analysis race other than white, history of weight loss, poor or moderate differentiation (P = 0.05), full thickness invasion (P = 0.02), positive lymph nodes (P < 0.01), Ivor Lewis esophagectomy (P = 0.02), intraoperative blood transfusion (P = 0.01), and tumor location in the upper or middle third in node-positive patients (P = 0.02) were associated with a poorer survival. Adjuvant therapy improved survival for patients with positive lymph nodes (P < 0.01). In multivariate analysis positive lymph nodes, tumor location, intraoperative blood transfusion, and adjuvant therapy were independent predictors of survival. To optimize survival esophagectomy for esophageal carcinoma should be performed without blood transfusion, and node-positive patients should receive multimodal therapy. PMID- 11893105 TI - Distal pancreatectomy: does the method of closure influence fistula formation? AB - The appropriate closure of the pancreatic remnant after distal pancreatectomy is still debated. Suture techniques, stapled closure, and pancreaticoenteric anastomosis all have their supporters. In this study we have reviewed our data from distal pancreatectomy to determine whether the type of remnant closure or underlying pathologic process had any relation to postoperative fistula formation. We performed a retrospective chart review of patients undergoing distal pancreatectomy at our institution between 1993 and 2001. The charts were reviewed for morbidity and mortality. These were then related to the type of closure of the pancreatic stump. From 1993 to 2001 a total of 86 patients underwent distal pancreatectomy. Data were available on 85 patients. Indications for surgery were pancreatic tumor (69%), pancreatitis (14%), trauma (7%), and extra pancreatic disease (9%). Pancreatic fistula occurred in 14 per cent (N = 12), intra-abdominal abscess in 8 per cent (N = 7), and wound infection in 2 per cent (N = 2). There was no mortality in the series. The incidence of pancreatic fistula formation was not related to method of closure of the pancreatic remnant nor to the underlying pathologic process. Postoperative pancreatic fistulas will close spontaneously even without total parenteral nutrition. PMID- 11893106 TI - Evaluation of vascular injury in penetrating extremity trauma: angiographers stay home. AB - The debate over the use of diagnostic angiography (DA) to exclude arterial injury in penetrating extremity trauma (PET) continues. This review evaluates our current protocol for PET and identifies indications for DA. Patients presenting to our urban Level I trauma center between January 1997 and September 2000 with PET were included. Demographic data, emergency department (ED) course, and patient follow-up were reviewed. ED evaluation directed by physical examination (PE) included Doppler pressure indices (DPI) and DA if indicated. A total of 538 patients had PET injuries. Twenty (4%) patients with hard signs of vascular injury were taken to the operating room. Ninety-one (17%) patients without vascular compromise underwent operative procedures or were admitted for other injuries. One hundred twenty-three (23%) patients with nonproximity wounds were discharged. Four DAs were performed for abnormal DPI with no change in management. Three hundred patients with a negative PE and normal DPI were discharged from the ED. Follow-up was available on 51 per cent of these patients (range 1-49 months) with no missed injuries identified. We conclude that PE with DPI is an appropriate way to identify significant vascular injuries from PET. Patients with normal PE and DPI can be safely discharged. DA is only indicated for asymptomatic patients with abnormal DPI. PMID- 11893107 TI - Carotid artery restenosis: an ongoing disease process. AB - Recurrence of carotid artery stenosis after primary endarterectomy is a well known entity. The treatment and optimal management of the disease process, however, is a matter of ongoing debate. We retrospectively reviewed carotid endarterectomies for recurrent disease performed at a community hospital over the past 21 years to evaluate the outcome of surgical intervention. Eighty-two recurrences occurred in 1648 carotid endarterectomies. Females had a slightly higher recurrence rate as compared with males, and the majority of patients had risk factors in the form of hypertension, peripheral vascular disease, or cigarette smoking. All endarterectomies were repaired with a patch angioplasty by either a vein or a prosthetic graft. One patient died secondary to complications of coronary artery disease. None of the patients developed any postoperative neurological event or permanent nerve damage. A subgroup of 11 patients with recurrent carotid artery stenosis with contralateral occlusion underwent 14 endarterectomies with no neurological complications. In conclusion occlusive carotid disease is an ongoing phenomenon, and continued surveillance is recommended. Surgical treatment of recurrent disease is a safe option. Endarterectomies for recurrent carotid disease in the presence of contralateral occlusion can be performed safely. PMID- 11893108 TI - Outcome analysis of pancreaticoduodenectomy at a community hospital. AB - There is an ongoing debate about the proposed regionalization of pancreaticoduodenectomies. The purpose of our study is to demonstrate that good outcomes can be achieved in a well-managed low-volume community hospital. We retrospectively analyzed pathologic findings, morbidity, mortality, and one-year survival in 32 patients who underwent pancreaticoduodenectomy at Providence Hospital over a 10-year period and compared these results with data collected at Johns Hopkins, and the Mayo Clinic. The patients had a mean age of 68.5 +/- 2.96 years; 56.3 per cent were female and 71.9 per cent were white. Overall in our series 90.6 per cent of specimens were found to be malignant, which is statistically higher than the 68 per cent at Johns Hopkins (P = 0.013) and not significantly different from Mayo Clinic (76%). The 30-day mortality rate at Providence Hospital was 3.1 per cent, which is not statistically different from Johns Hopkins (1.3%) and Mayo Clinic (3.6%). One-year survival rate at Providence Hospital was 59.4 per cent, which is significantly different from 79 per cent at Johns Hopkins (P = 0.016). The one-year survival rate at Providence Hospital is higher than an approximately 50 per cent average reported nationally. The postoperative complication rate was 62.5 per cent; the most common complication was delayed early gastric emptying (28.1%). A statistical difference in morbidity exists between Providence Hospital and Johns Hopkins (P = 0.027) but not between Providence Hospital and Mayo Clinic (46%). The higher rate of malignant disease treated in the population at Providence Hospital may contribute to a higher complication rate and lower one-year survival rate than the reported rates at Johns Hopkins because of the poorer health of cancer patients. However, statistical analysis of mortality rates for pancreaticoduodenectomy at Providence Hospital show no difference from mortality rates at Johns Hopkins and Mayo Clinic. Therefore in low-volume community hospitals pancreaticoduodenectomy can be performed safely as evidenced by a comparable low mortality rate and a high one-year survival rate. PMID- 11893109 TI - Evaluation of pulmonary arterial catheter parameters utilizing intermittent pneumatic compression boots in congestive heart failure. AB - The use of intermittent pneumatic compression boots to reduce the risk of deep venous thrombosis is contraindicated in patients with congestive heart failure (CHF) due to a theoretical increase in venous return to the heart and exacerbation of heart failure. This study evaluates intermittent pneumatic compression effects on pulmonary artery catheter parameters in CHF patients. We conducted a prospective within-patient study of CHF patients monitored by pulmonary artery catheterization. Hemodynamic variables were assessed with and without the use of intermittent pneumatic compression boots. A sample size of 18 patients was calculated a priori to obtain an 80 per cent power to detect a mean difference of 10 per cent. Twenty patients were studied; no patient suffered hemodynamic instability during the application of pneumatic compression; no statistically significant change in any hemodynamic parameters was noted. A trend toward decreasing mean arterial blood pressure (P = 0.057), pulmonary artery wedge pressure (P = 0.065), and systemic vascular resistance (P = 0.08) was observed. None were clinically significant. The application of intermittent pneumatic compression to the feet of patients in CHF does not significantly alter central hemodynamic parameters in CHF patients. This study suggests that intermittent pneumatic compression may be used in CHF patients for venous thromboembolic risk reduction. PMID- 11893110 TI - Laparoscopic ventral hernia repair: are there comparative advantages over traditional methods of repair? AB - Recent studies have noted advantages of laparoscopic over open repair of ventral hernias. Because few reports have involved comparison with traditional repair we report a comparison between laparoscopic and open approaches. We retrospectively reviewed the records of patients undergoing ventral hernia repair over a 28-month period. Patients were grouped into three categories: laparoscopic repair with mesh, open repair with mesh, and open repair without mesh. There were 295 ventral hernia repairs and there was no difference in age, gender, operative complications, or hospital stay between the groups. Mesh and defect size was greater in the laparoscopic group. The overall postoperative complication rate was greater in the open group with mesh. Yet when specific wound complications were analyzed there was no difference between the groups. Furthermore a death occurred in the laparoscopic group from an unrecognized bowel injury. The recurrence rate was greatest in the open repair without mesh group. Finally hospital cost was greatest in the laparoscopic group and third-party reimbursement was better for the open techniques. We were unable to demonstrate a significant advantage to laparoscopic ventral hernia repair. Although many patients with large fascial defects were well served with this approach it may not be a better option for these patients. PMID- 11893111 TI - Fast track video-assisted thoracic surgery. AB - Video-assisted thoracic surgery (VATS) has been advocated as one of the primary diagnostic modalities for suspicious pulmonary nodules and diffuse interstitial lung disease. The aim of our study was to evaluate the cost and safety of VATS lung wedge resection(s) as an "overnight" hospital admission. We retrospectively reviewed all 37 charts of patients who underwent VATS wedge resections for these indications from August 1999 to April 2001. There was a slight female predominance with mean age of 56.8 years (range 33-88). Eighteen patients had interstitial disease and 19 patients had pulmonary nodules. The duration of chest tube drainage was one day in the majority (92%). Length of hospital admission was overnight in 70 per cent whereas 22 per cent remained two days. This latter group from the earlier period of the trial had characteristics identical to those of an overnight stay. This creates a potential overnight stay in 87 per cent. Five complications occurred in three patients, which extended the length of stay. No mortality was reported. The overall hospital charges for the overnight-stay VATS were nearly half the charges for the open thoracotomy counterpart. Diagnostic VATS wedge biopsy is a cost effective and safe procedure allowing an overnight hospital stay in the majority of cases. PMID- 11893112 TI - The impact of rural clinical placement on student nurses' employment intentions. AB - Commonwealth Government health policy and professional organisations have indicated that successful recruitment and retention strategies are crucial to address the shortage of health professionals in rural and remote areas. This research study aimed to evaluate the effectiveness of a Clinical Placement Support Scheme for nursing students as a recruitment strategy for rural and remote health care services, and to develop an increased awareness of the employment opportunities available in these areas. The population consisted of final year Bachelor of Nursing students enrolled in either a rural or metropolitan clinical placement in 2000. A pre-post test survey design was used. Analysis of pre- and post-test data found a 12% increase (to 89%) in the number of students intending to seek employment in a rural setting, compared to a 5% increase (to 46%) in students who undertook a metropolitan placement. One-third of the students who chose a rural placement had no previous experience of a rural lifestyle and over half of these students indicated their intention to work in a rural setting following their clinical placement. These results support the theory that undergraduate rural clinical experience can have a positive influence on the recruitment of health professionals to rural areas. PMID- 11893113 TI - The postmodern heart: a discourse analysis of a booklet on pacemaker implantation. AB - This paper provides a deconstructive reading of an education booklet aimed at patients who may be scheduled for permanent cardiac pacemakers, and argues that pacemaker implantation is being constructed as a routine, uncomplicated procedure. The way language and image in the booklet convey a dominant preferred perspective on pacemaker implantation will be questioned. Some of the major issues explored are: the patient is stereotyped as a male patriarch; the fundamental right of informed choice is omitted; the ownership of decision-making is given to the physician; and the cardiac pacemaker is portrayed as remarkable and trouble-free. Implications for the meaning of these dominant perspectives for nursing practice are explored. PMID- 11893114 TI - From apprentices to academics: are nurses catching up? AB - In comparison to other disciplines, Australian nursing has only come relatively lately to academia. Traditionally, academic qualifications were not viewed as necessary for nurses. The movement of nursing education to the tertiary sector has seen many changes from the traditional apprenticeship model and the characteristics of nurse-academics reflect these. The researchers identified changes that have occurred in the last five years in nurse-academics' qualifications, academic rank and links between them. It is clear that the goalposts for nurse academics have moved, with a master's degree now standard for Lecturer Level B and a doctorate for Level D. Other findings show a strong link between movement (transfer), increased qualifications and promotion. Females were more likely than males to have increased their qualifications and to be promoted. In terms of academic qualifications in the whole system, female nurse-academics have caught up with their counterparts in the former CAE sector. Male nurse academics have parity with female nurse-academics but not with males in the system generally. The study shows the great strides that Australian nurse academics have made in the five years preceding the end of the old millenium but illustrates that they and female academics generally have not yet caught up to their male colleagues. PMID- 11893115 TI - Nursing and the 21st century: what's happened to leadership? AB - This paper has stemmed from several conversations between the authors over recent times about the current state of leadership in nursing. The authors believe a lack of effective leadership has much to do with the times in which we live. As a result, this paper explores leadership against a backdrop of social and political change and seeks to revise the concept in light of 21st century developments. The demise of leadership is discussed and ideas about the emergence of new leadership brought to light. Finally the need for nurses and nursing to shake off the depressive effects of economic rationalism, join forces and unite for a cause is recommended. PMID- 11893116 TI - Nurses' views on competency indicators for Australian nursing. AB - This paper reports on a project commissioned by the Australian Nursing Council Inc that sought to develop an approach to the maintenance of continuing competence in nursing broadly acceptable to nurses in all states and territories. This project involved extensive consultation with nurses, consumers and key stakeholders on appropriate competence indicators. Findings suggest that a majority of nurses support the development of competence indicators but most are confused about the nature of competence. PMID- 11893117 TI - Health promotion in the changing face of the hospital landscape. PMID- 11893118 TI - Our interest is swelling: a collaborative venture to establish the world's first free public screening for lymphoedema. AB - This article describes the collaborative processes involved in the implementation of a free public health screening program for people at risk of lymphoedema following the removal of lymph nodes during surgery to control breast, prostate and other cancers, or injury. The planning phase of the program is described with emphasis on the need to secure a well situated venue, the commitment of a cohort of key health professionals, service club and lay volunteers, and the need to carefully target and publicise the event widely. The implementation phase requires careful consideration of the physical layout of the event, the direction and management of the flow of human traffic, information and equipment requirements, and recognition that screening programs place people in vulnerable positions. Effective communication skills are essential, as is a knowledge of where people can be referred should the need arise. A budget is provided together with discussion regarding the success of the program and recommendations for future consideration such as the need to target men to attend screening and for long term follow up of the outcomes. PMID- 11893119 TI - Scholars we are! PMID- 11893120 TI - Ageing of the nursing workforce: implications for acute care services. PMID- 11893121 TI - Targetting out of the workforce nurses: a promising local recruitment strategy. AB - A successful pilot program to attract and orient 'out of the workforce' nurses to a hospital outreach acute and post acute care service is described. Three new staff members attended a one-month education program prior to commencing supervised clinical activity. Survey and focus group methods were used to evaluate the initiative. Principal lessons learned were that preceptoring and preceptor support is central to a positive outcome for new and existing staff. We also learned that a tendency to underestimate the complexity of the post acute care nursing role in the initial advertising was unhelpful. PMID- 11893122 TI - Detection of a gabor patch superimposed on an illusory contour. AB - Interactions between visual stimuli have been found to be specific to the spatial frequency, orientation and phase of the interacting stimuli. We asked if there are any interactions between luminance-defined Gabor patches and Kanizsa-type illusory contours. In psychophysical experiments we studied whether induction of a vertical illusory line affects detection thresholds for a Gabor patch superimposed on this line and whether these effects depend on the orientation, spatial frequency and phase of the Gabor elements. Employing a 2AFC method with a staircase procedure we measured contrast detection thresholds and varied the orientation, spatial frequency and phase of the test Gabor patch and the separation between the two pacmen in four experimental series. The results show that in a situation where the two inducers generate perception of an illusory line, the contrast detection of the Gabor patch is facilitated relative to a control condition where the rotated pacmen do not induce illusory contours. This facilitation was more pronounced for test Gabor signals that were collinear to the illusory line, but the observer's performance was not altered by changes in the spatial frequency or phase of the Gabor stimuli. With increasing spatial separation of the two pacmen (and, consequently, with a decreasing support ratio), the difference between performance in the test and control conditions diminished. From the data obtained we cannot infer that we have measured some neural interactions between Gabor patches and Kanizsa-type illusory contours, and nor can we draw a unique conclusion about what causes the facilitation of detection of the test Gabor patch in the experimental situation that allows induction of the illusory line. We discuss possible mechanisms of the facilitation, such as contextual influences or a reduction of uncertainty about spatial location of the test Gabor patch. PMID- 11893123 TI - Contrast discrimination and choice reaction times at near-threshold pedestals. AB - Choice reaction times (CRTs) to contrast differences were measured and compared with contrast increment thresholds obtained from concurrently measured psychometric functions at pedestal contrasts in the vicinity of detection threshold. Contrast discrimination functions had a classical dipper shape. The main finding was that CRTs were shorter at low pedestal contrasts but longer at higher pedestal contrasts compared to detection, reflecting the behaviour of increment thresholds. Even when equalized for response accuracy, CRTs varied with pedestal contrast in a similar manner to the contrast increment thresholds. The finding that CRTs and contrast increment thresholds depended on pedestal contrast in a similar manner suggests that both share a common origin. This common origin is proposed to lie in the variability of the sensory effect which determines the variability of the information accumulation process, which in turn affects the response criterion and contrast increment thresholds. At low pedestals, a decrease in variability lowers thresholds and results in a lower response criterion, thereby accelerating reaction times. At high pedestals, increasing signal-dependent noise inflates variability and thus raises thresholds and the response criterion, which results in slower CRTs. PMID- 11893124 TI - Occlusion junctions do not improve stereoacuity. AB - Occlusion geometry gives rise to interocular shifts in the positions of binocularly viewed contour junctions. Since these shifts do not give rise to normal binocular disparities, they have been called 'pseudodisparities'. Previous work has shown that the unmatched contour segments of a partially occluded contour at occlusion junctions can be used to recover the geometry of the occluding surface through the construction of 'illusory' contours. Here, experiments were performed to determine whether such junction shifts could enhance stereoscopic depth detection when the relative disparity between the contours was below threshold. Our results showed that stereoscopic depth detection does not improve when pseudodisparity is present. We conclude that the visual system is less sensitive to pseudodisparity than to conventional disparity information. We suggest that the primary role of pseudodisparity is to overcome conditions of camouflage. PMID- 11893125 TI - Characterizing visual performance fields: effects of transient covert attention, spatial frequency, eccentricity, task and set size. AB - We investigated whether transient covert attention would differentially affect 'performance fields' (shape depicted by percent correct performance at particular locations in the visual field) for orientation discrimination, detection and localization tasks, while manipulating a number of visual factors. We found that although attention improved overall performance, it did not affect performance fields. Two patterns were observed regardless of the presence of a local post mask, the stimulus orientation, or the task. A horizontal-vertical anisotropy (HVA) became more pronounced as spatial frequency, eccentricity and set size increased. A vertical meridian asymmetry (VMA) became more pronounced as spatial frequency and eccentricity increased. We conclude that performance fields are determined by visual, rather than by transient attentional, constraints. PMID- 11893126 TI - The role of vergence in the perception of distance: a fair test of Bishop Berkeley's claim. AB - Binocular eye movements were measured while subjects perceived the wallpaper illusion in order to test the claim made by Bishop Berkeley in 1709 that we perceive the distance of nearby objects by evaluating the vergence angles of our eyes. Four subjects looked through a nearby fronto-parallel array of vertical rods (28-35 cm away) as they binocularly fixated a point about 1 meter away. The wallpaper illusion was perceived under these conditions, i.e. the rods appeared farther away than their physical location. We found that although binocular fixation at an appropriate distance was needed to begin perceiving the wallpaper illusion (at least for naive observers), once established, the illusion was quite robust in the sense that it was not affected by changing vergence. No connection between the apparent localization of the rods and vergence was observed. We conclude that it is unlikely that vergence, itself, is responsible for the perceived distance shift in the wallpaper illusion, making it unlikely that vergence contributes to the perception of distance as Bishop Berkeley suggested. We found this to be true even when vergence angles were relatively large (more than 2 deg), the region in which the control of vergence eye movements has been shown to be both fast and effective. PMID- 11893127 TI - Contrast dependency of VEPs as a function of spatial frequency: the parvocellular and magnocellular contributions to human VEPs. AB - The present study investigated the contrast dependency of visual evoked potentials (VEPs) elicited by phase reversing sine wave gratings of varying spatial frequency. Sixty-five trials were recorded for each of 54 conditions: 6 spatial frequencies (0.8, 1.7, 2.8, 4.0, 8.0 and 16.0 c deg(-1)) each presented at 9 contrast levels (2, 4, 8, 11, 16, 23, 32, 64 and 90%). At the lowest spatial frequency, the waveform contained mainly one peak (P1). For spatial frequencies up to 8 c deg(-1), P1 had a characteristic magnocellular contrast response: it appeared at low contrasts, increased rapidly in amplitude with increasing contrast, and saturated at medium contrasts. With increasing spatial frequency, an additional peak (N1) gradually became the more dominant component of the waveform. N1 had a characteristic parvocellular contrast response: it appeared at medium to high contrasts, increased linearly in amplitude with increasing contrast, and did not appear to saturate. The data suggest the contribution of both magnocellular and parvocellular responses at intermediate spatial frequencies. Only at the lowest and highest spatial frequencies tested did magnocellular and parvocellular responses, respectively, appear to dominate. PMID- 11893128 TI - Identifying patient preoperative risk factors and postoperative adverse events in administrative databases: results from the Department of Veterans Affairs National Surgical Quality Improvement Program. AB - BACKGROUND: The Department of Veterans Affairs (DVA) National Surgical Quality Improvement Program (NSQIP) employs trained nurse data collectors to prospectively gather preoperative patient characteristics and 30-day postoperative outcomes for most major operations in 123 DVA hospitals to provide risk-adjusted outcomes to centers as quality indicators. It has been suggested that routine hospital discharge abstracts contain the same information and would provide accurate and complete data at much lower cost. STUDY DESIGN: With preoperative risks and 30-day outcomes recorded by trained data collectors as criteria standards, ICD-9-CM hospital discharge diagnosis codes in the Patient Treatment File (PTF) were tested for sensitivity and positive predictive value. ICD-9-CM codes for 61 preoperative patient characteristics and 21 postoperative adverse events were identified. RESULTS: Moderately good ICD-9-CM matches of descriptions were found for 37 NSQIP preoperative patient characteristics (61%); good data were available from other automated sources for another 15 (25%). ICD-9 CM coding was available for only 13 (45%) of the top 29 predictor variables. In only three (23%) was sensitivity and in only four (31%) was positive predictive value greater than 0.500. There were ICD-9-CM matches for all 21 NSQIP postoperative adverse events; multiple matches were appropriate for most. Postoperative occurrence was implied in only 41%; same breadth of clinical description in only 23%. In only four (7%) was sensitivity and only two (4%) was positive predictive value greater than 0.500. CONCLUSION: Sensitivity and positive predictive value of administrative data in comparison to NSQIP data were poor. We cannot recommend substitution of administrative data for NSQIP data methods. PMID- 11893129 TI - The frequency and effects of cytochrome P450 (CYP) 2C9 polymorphisms in patients receiving warfarin. AB - BACKGROUND: Warfarin sodium (warfarin) is commonly prescribed in surgical practice. Warfarin use is complicated by an unpredictable dose response that may be due in part to genetically determined differences in metabolic capacity. To better understand the interaction between genotype and response to warfarin therapy, we determined the frequency and functional effects of polymorphisms of the predominant cytochrome P450 subfamily responsible for warfarin metabolism (eg, CYP2C9) in an ethnically defined U.S. patient population. DESIGN: Patients requiring chronic anticoagulation with warfarin sodium (warfarin) were recruited over an 11-month period (June 1999 through May 2000) from the inpatient and outpatient divisions of a tertiary care medical center in this prospective observational study. Clinical and demographic information was collected and CYP2C9 genotype was determined. RESULTS: One hundred fifty-three patients receiving warfarin therapy for at least four weeks and comprising two ethnic groups [33 African Americans (22%) and 120 Caucasians (78%)] were genotyped. The mean weekly warfarin dose (+/-SEM) for all patients [36.9 (+/- 1.5) mg] was not influenced by gender [85 males (56%), 68 females (44%)] or ethnicity (p>0.05 for both), but was significantly affected by age (p = 0.006 for weight adjusted warfarin dose). The frequencies of CYP polymorphisms were as follows: 2C9*2 (24/153) 15.7%, 2C9*3 (23/153) 15.0%. There were no gender differences in polymorphism frequency (CYP2C9*2 frequency = (13/ 85) 15.3% in males, (12/68) 17.6% in females, p=0.74; CYP2C9*3 frequency = (15/85) 17.6% in males and (8/68) 11.8% in females, p = 0.38). CYP polymorphisms were much less common in African Americans than Caucasians [(5/33) 15.2% versus (47/120) 39.2%, respectively p = 0.05)]. Patients with CYP polymorphisms (2C9*2, 2C9*3) had significantly lower warfarin doses compared to patients with wild-type genotypes [30.6 (+/- 2.5) mg versus 40.1 (+/- 1.7) mg, p = 0.0021] . Stepwise backward regression analysis suggested a moderate ability to predict warfarin dose based on CYP genotype (r2 = 0.26), p < 0.01). CONCLUSIONS: CYP2C9 polymorphisms are common, associated with significant reductions in warfarin dose, and partly account for interpatient variability in warfarin sensitivity. As interactions between genetic factors and other variables that influence warfarin effect are more completely understood, CYP analysis may prove a useful adjunct for increasing the safety and efficacy of this agent. PMID- 11893130 TI - Thyroglossal duct: a review of 55 cases. AB - BACKGROUND: Thyroglossal duct remnants are the most common midline neck masses in childhood but can be found in adults and the elderly. Sistrunk's procedure, with dissection of the tract and removal of the hyoid bone, is accepted as the main operation of choice. STUDY DESIGN: Fifty-five patients were treated from January 1994 to November 2000, and these were studied. There were 29 men and 26 women, with a median age of 17 years. Diagnosis was clinical, with 13 cases of fistula and 42 of cyst. Size varied from 1.0 to 4.0 cm, with an average of 2.5 cm. Six patients presented with local abscess. RESULTS: All the patients underwent Sistrunk's procedure. Serum collection occurred in three patients as complication. In one patient papillary carcinoma was identified in the cyst. Total thyroidectomy was not performed. There was only one recurrence, managed with a second operation. CONCLUSIONS: We concluded that the diagnosis of thyroglossal duct is clinical. Sistrunk's procedure carries low rates of complications (9.08%) and recurrence (1.82%). Antibiotic therapy is avoidable as a rule and hospital stay is short. PMID- 11893131 TI - Does breast tumor location influence success of sentinel lymph node biopsy? AB - BACKGROUND: Controversy exists regarding the influence of sentinel lymph node (SLN) mapping technique or patient variables on the success rate of SLN mapping. We undertook a prospective study in a single institution series to evaluate multiple variables that could adversely affect SLN identification rates. STUDY DESIGN: Data were collected on 174 patients who underwent 177 SLN mapping procedures followed by axillary dissection from October 1996 through January 2000. Patient demographics, body mass index (BMI), biopsy method, tumor size, palpability, and location were recorded. SLNs were identified by blue dye only (n = 31), Tc-99m sulfur colloid only (n = 34), or combined techniques (n = 112). Data were analyzed by logistic regression analysis and expressed as the probability of failure to map the SLN. RESULTS: SLNs were identified successfully in 150 of 177 procedures (85%) with a false negative rate of 3.7%. Mapping success reached 93% using combination blue dye and isotope. Variables found to adversely affect SLN mapping success and the odds ratio of failure (OR) included lower inner quadrant (LIQ) location (OR 35.6), blue dye only (OR 42.4), BMI >30 and upper outer quadrant (UOQ) location (OR 14.6), and nonpalpable UIQ location (OR 25). LIQ location adversely affects mapping success independent of technique, tumor size, or obesity. Obesity and nonpalpability were adverse factors when tumors were located in the UOQ and UIQ, respectively. Age, biopsy technique, and tumor diameter did not affect SLN mapping success. CONCLUSIONS: SLN mapping success is influenced by technique and tumor location, with best results achieved using combined techniques and for lesions located in quadrants other than the LIQ. Obesity and tumor palpability influence success in the context of tumor location. PMID- 11893132 TI - Early complications after Ivor Lewis subtotal esophagectomy with two-field lymphadenectomy: risk factors and management. AB - BACKGROUND: Esophageal resection represents a major surgical and physiologic insult carrying major morbidity and mortality. We present the results of esophagectomy in a specialist unit with emphasis on early complications and their management. STUDY DESIGN: From January 4, 1990 through January 6, 2000, 228 patients have undergone Ivor Lewis subtotal esophagectomy with two-field lymphadenectomy for malignancy under the care of one surgeon. The median age was 64 years (range 39 to 77 years), with a male to female ratio of 2.3:1 and a predominance of adenocarcinoma (n = 146) compared with squamous cell carcinoma (n = 75) and other tumors (n = 7). Detailed prospective data were collected on preoperative status, operative parameters, and postoperative complications. RESULTS: Median ICU stay was 1 day (range 1 to 47 days) and the median postoperative hospital stay in patients surviving surgery (n= 219) was 13 days (range 9 to 159 days). There were 119 separate postoperative complications occurring in 45% of patients (102 of 228), comprising predominantly pulmonary morbidity. Major respiratory complications (17%) were significantly associated with poor preoperative spirometry (p = 0.002) and a history of smoking (p = 0.03). Seven percent of patients (16 of 228) suffered cardiovascular or thromboembolic complications. Major surgical complications occurred in 10% of patients (22 of 228) including mediastinal leaks in 4%. Isolated anastomotic leaks (2%) were successfully treated conservatively in all cases; extensive leaks from ischemic gastric conduits (1%) or gastrotomy dehiscence (1%) underwent further exploration and either local repair or resection and exclusion. Reoperation for hemostasis was required in 3% (6 of 228) and only 1% of patients (2 of 228) developed chyle leaks. Thirty-day mortality was 2%, rising to 4% for in-hospital mortality. The nine fatalities were significantly older (p = 0.02) than those who survived and 67% (6 of 9) had suffered primary surgical complications. CONCLUSIONS: Overall morbidity after radical esophagectomy is high, but early recognition and aggressive management of complications can minimize subsequent mortality. Concentration of facilities and surgical expertise in specialist units together with more careful patient selection can decrease mortality further. PMID- 11893133 TI - Factors associated with conversion to laparotomy in patients undergoing laparoscopic appendectomy. AB - BACKGROUND: Laparoscopic appendectomy (LA) has been increasingly adopted for its advantages over the open technique, but there is a possibility of conversion to open appendectomy (OA) if complications occur or the extent of inflammation prohibits successful dissection. This study aimed to identify the preoperative predictors for conversion from laparoscopic to open appendectomy. STUDY DESIGN: Medical records of 705 consecutive patients who underwent surgery for suspected appendicitis were reviewed retrospectively. LA was attempted in 595 patients by 25 different surgeons. Factors evaluated were age, gender, body mass index, previous abdominal surgery, previous appendicitis attack, pain, nausea, vomiting, fever, duration of symptoms, local or diffuse tenderness, leukocyte count and surgeon's experience in LA. RESULTS: Conversion to OA occurred in 58 patients (9.7%). The most common reason for conversion was dense adhesions due to inflammation, followed by localized perforation and diffuse peritonitis. Based on 261 patients evaluated by CT scan preoperatively, significant factors in the final multivariate analysis associated with conversion to OA were age > or = 65 [Odds ratio (OR) = 3.78, 95% CI:1.11-12.84], diffuse tenderness on physical examination (OR = 11.32, 95% CI: 1.32-96.62), and a surgeon with less experience in LA (< or = 10 operations, OR = 3.38, 95% CI:1.02-11.17). The presence of significant fat stranding associated with fluid accumulation, inflammatory mass or localized abscess in CT scan also significantly increased the possibility of conversion (OR = 5.60, 95% CI:2.48-12.65). CONCLUSIONS: Identifying the potential factors for conversion preoperatively may assist the surgeons in making decisions concerning the management of patients with appendicitis and in the judicious use of LA. PMID- 11893134 TI - Nonparasitic splenic cysts: pathogenesis, classification, and treatment. AB - BACKGROUND: Nonparasitic splenic cysts (NPSCs) are uncommon lesions of the spleen, many being reported in anecdotal fashion. Early classifications of this disorder have been based on the presence or absence of an epithelial lining, indicating either a congenital or traumatic etiology. This criterion has led to confusion and mistaken reporting because the lining alone is not a reliable criterion. STUDY DESIGN: Over a 28-year period, the author has observed and studied 23 patients with NPSC. Special attention has been given to the role of trauma in the history, the nature (or absence) of a cyst lining, the gross pathology, and the preferred method of treatment. RESULTS: NPSC present as lesions with a very characteristic gross appearance and lining. The trabeculated interior can be lined with epidermoid, transitional, or mesothelial epithelium. Desquamation of the lining can lead to a spurious diagnosis, but careful search usually discloses the lining remnant. Although most NPSC in this series were treated by open partial splenectomy, the more recent approach by laparoscopic techniques offers great promise. CONCLUSIONS: A new classification of NPSC is offered, based on characteristic gross findings. NPSC are of congenital origin, with a lining derived from mesothelium. Trauma does not play a primary role in pathogenesis. Cysts that are symptomatic or over 5 cm in diameter should be removed by partial splenectomy or near-total cystectomy "decapsulation," either by the open or laparoscopic approach. PMID- 11893136 TI - What's new in general surgery: trauma and critical care. PMID- 11893135 TI - Gynecologic abnormalities in surgically treated women with stage II or III rectal cancer. AB - BACKGROUND: The objective of this article is to review the incidence and management of gynecologic abnormalities in women undergoing surgery for rectal cancer. STUDY DESIGN: We performed a retrospective chart review utilizing the Johns Hopkins Tumor Registry and Pathology database. Eighty-six female patients who underwent abdominal surgery between 1985 and 1996 for Stage II or Stage III rectal cancer were identified. Data gathered included: patient demographics, history, intraoperative findings and complications, cancer stage and histology, adjuvant treatments, and followup. Specific attention was focused on the diagnosis, management, and followup of concurrent gynecologic problems. RESULTS: At the time of surgery, nineteen women (22%) had previously undergone hysterectomy and bilateral salpingo-oophorectomy. Of the remaining 67 patients, 25 (37%) were found to have gynecologic abnormalities at the time of surgery, 15 (22%) underwent adnexectomy or hysterectomy or both. Forty-two women (63%) had normal internal genitalia. Of the 61 peri- and postmenopausal women, nine underwent bilateral oophorectomy for therapeutic reasons. No prophylactic oophorectomies were performed in any of the patients. CONCLUSION: Incidental pathologic findings necessitating gynecological procedures are common in patients undergoing surgery for rectal cancer. These findings are frequently suboptimally assessed and managed in the pre-, intra-, and postoperative periods. Colorectal surgeons operating on women with Stage II and III rectal cancer should be cognizant of the high likelihood of identifying incidental gynecologic pathology and be prepared for definitive management of the pathology. The utilization of prophylactic oophorectomy in postmenopausal women undergoing surgery for rectal cancer is currently not optimal; preoperative discussion should address this option. PMID- 11893137 TI - Ethics and philosophy lecture: surgery...Is it an impairing profession? PMID- 11893138 TI - Overview of bariatric surgery. PMID- 11893139 TI - Management of dyspnea at the end of life: relief for patients and surgeons. PMID- 11893140 TI - Intravascular hemolysis from a Clostridium perfringens liver abscess. PMID- 11893141 TI - Endoscopically assisted "components separation technique" for the repair of complicated ventral hernias. PMID- 11893142 TI - An improved, inexpensive, quick, and easily learned technique for closure of all large abdominal trocar wounds after laparoscopic procedures. PMID- 11893143 TI - Managing pediatric solid organ injury. PMID- 11893144 TI - Risk factors for vancomycin-resistant Enterococcus (VRE) infection in colonized patients with cancer. AB - To determine the risk factors for vancomycin-resistant Enterococcus (VRE) infection in colonized patients with cancer, we conducted a case-control study. According to multivariate analysis, the only significant factors were neutropenia (< 500 cells/mm3) for more than 1 week and the use of oral vancomycin. Therefore, colonized neutropenic patients with cancer who have previously used oral vancomycin are most prone to VRE infection. PMID- 11893145 TI - Outbreak of burkholderia cepacia in the adult intensive care unit traced to contaminated indigo-carmine dye. AB - We report an unusual cluster of Burkholderia cepacia in patients. Environmental cultures identified indigo-carmine dye used in enteral feeding as the reservoir. Compared with the controls, the cases were significantly more likely to have received tube feedings tinted with this dye. This outbreak was terminated with the removal of the dye from hospital inventory. PMID- 11893146 TI - The cost of antibiotic resistance: effect of resistance among Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudmonas aeruginosa on length of hospital stay. AB - To assess the effect of antimicrobial resistance on length of hospital stay, a case-control study compared infections due to four nosocomial pathogens. Significantly increased lengths of stay were associated with infections due to methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamase producing Klebsiella pneumoniae, and carbapenem-resistant Acinetobacter baumannii or Pseudomonas aeruginosa. Infections with resistant pathogens are associated with prolonged hospitalization. PMID- 11893147 TI - Nasal carriage of methicillin-susceptible and methicillin-resistant Staphylococcus aureus among paramedics in the Sedgwick County emergency medical service in Wichita, Kansas. PMID- 11893148 TI - Efficacy of alcohol-based hand sanitizers against fungi and viruses. PMID- 11893149 TI - Nosocomial outbreak of Kluyvera cryocrescens bacteremia. PMID- 11893150 TI - Would active surveillance cultures help control healthcare-related methicillin resistant Staphylococcus aureus infections? PMID- 11893151 TI - The best hospital practices for controlling methicillin-resistant Staphylococcus aureus: on the cutting edge. AB - OBJECTIVE: A performance improvement task force of Rhode Island infection control professionals was created to develop an epidemiologic model of statewide consistent infection control practices that could reduce the spread of methicillin-resistant Staphylococcus aureus (MRSA). DESIGN: This model encompasses screening protocols, isolation techniques, methods of cohorting positive patients, decolonization issues, postexposure follow-up, microbiology procedures, and standardized surveillance methodologies. These "best practice guidelines" include three categories of recommendations that define priority levels based on the availability of scientific data. SETTING: From 1995 through 2000, several Rhode Island hospitals experienced a fivefold increase in nosocomial acquisition of MRSA PARTICIPANTS: Rhode Island infection control professionals are a highly interactive group in the unique position of sharing patients and ultimately experiencing similar trends and problems. INTERVENTION: The task force collaborated on developing the best hospital infection control practices to prevent and control the spread of MRSA in Rhode Island. RESULTS: The task force met with local infectious disease physicians and representatives from the Rhode Island Department of Health, the Hospital Association of Rhode Island, and Rhode Island Quality Improvement Partners. Discussions identified numerous and diverse MRSA control practices, issues of consensus, and approaches to resolving controversial methods of reducing the spread of MRSA. The guidelines regarding the best hospital practices for controlling MRSA were finalized 8 months later. CONCLUSION: These guidelines were distributed to all chief executive officers of Rhode Island hospitals by the Rhode Island Department of Health in December 2001. They were issued separate and apart from any regulations, with the intent that hospitals will adopt them as best hospital practices in an attempt to control MRSA. PMID- 11893152 TI - An outbreak of Staphylococcus aureus in a pediatric cardiothoracic surgery unit. AB - OBJECTIVE: To investigate an outbreak of Staphylococcus aureus surgical-site infections. DESIGN: Case-control study. SETTING: Pediatric cardiothoracic surgery service of a tertiary-care university medical center. METHOD: Molecular typing was used to identify healthcare workers who carried the epidemic strain. RESULTS: Three children acquired surgical-site infections caused by a single strain of S. aureus. Fourteen (25%) of the staff members in the operating room and 17 (11%) on nursing units carried the epidemic strain (P = .01). A case-control study identified 4 healthcare workers who were associated statistically with the outbreak, 2 of whom (a cardiothoracic surgeon and a perfusionist) carried the epidemic strain in their nares. The surgeon also carried the epidemic strain on his hands. Each staff member who carried the epidemic strain was treated with mupirocin; those carrying the strain on their hands were required to wash their hands with chlorhexidine. The surgeon was not allowed to perform surgery until 2 of his hand cultures did not grow S. aureus. CONCLUSIONS: Only three children were infected with the epidemic strain, but it was disseminated widely among staff who cared for children who underwent cardiothoracic surgery. No additional cases were identified after staff members who carried the epidemic strain were decolonized. Both classic epidemiologic methods and molecular typing techniques were necessary to identify the source and extent of this outbreak. PMID- 11893153 TI - The effectiveness of influenza vaccine against influenza a (H3N2) virus infections in nursing homes in Niigata, Japan, during the 1998-1999 and 1999-2000 seasons. AB - OBJECTIVE: To evaluate the effectiveness of influenza vaccines against influenza like illness (ILI) among nursing home residents. DESIGN: Prospective, nonrandomized, cohort study. SETTING: Nine nursing homes during the 1998-1999 influenza season and 11 nursing homes during the 1999-2000 influenza season in Niigata Prefecture, Japan. PARTICIPANTS: Six hundred ninety-nine residents and 440 healthcare workers (HCWs) during the first season, and 930 residents and 517 HCWs during the second season, with vaccination rates ranging from 0% to 97.7%. RESULTS: Overall, ILI decreased from 24.3% during the 1998-1999 season to 8.8% during the 1999-2000 season. Multivariate analysis adjusted for several factors, including gender, age, underlying diseases, and resident and HCW vaccination rates, failed to demonstrate clear individual protection of residents (relative risk [RR], 1.42; P = .2 for the first season; RR, 0.95; P = .9 for the second season). However, vaccination rates of 60% or greater for residents and HCWs reduced the risk of ILI, and also could prevent outbreaks during the 2 seasons. Highly impaired activities of daily living and chronic respiratory diseases were significantly associated with increased ILI. CONCLUSIONS: A high vaccination rate for both residents and HCWs may reduce the risk of ILI and institutional outbreaks in nursing homes. PMID- 11893154 TI - A quality management project in 8 selected hospitals to reduce nosocomial infections: a prospective, controlled study. AB - OBJECTIVE: To reduce the number of nosocomial infections (NIs) in surgical patients by a quality management approach. DESIGN: Prospective, controlled study in 8 medium-sized hospitals during a 26-month period. SETTING: Four study hospitals and 4 control hospitals. METHODS: In two 10-month intervention periods, 4 external physicians introduced quality circles and ongoing surveillance in the 4 study hospitals. There were three 8-week observation periods in all 8 hospitals with the same physicians before, during, and after the intervention periods. RESULTS: During the first observation period, almost identical overall incidence densities were found for the study hospitals and the control hospitals. During the course of the study, the overall incidence density decreased significantly in the study hospitals (risk ratio [RR], 0.74; 95% confidence interval [CI 95], 0.59 to 0.94) and nonsignificantly in the control hospitals (RR, 0.90; CI 95 0.70 to 1.16). With the use of a Cox regression model to evaluate the impact of the intervention periods while taking into account the distribution of risk factors for NI in both groups, a significant risk reduction (RR, 0.75; CI 95, 0.58 to 0.97) was observed after the first intervention period when comparing study and control hospitals. At the end of the study (ie, after the second intervention period), the difference between the study hospitals and the control hospitals was not significant (RR, 0.78; CI 95, 0.60 to 1.01). This was due to no further improvement at the end of the study in the study hospitals and a decrease in the control hospitals. CONCLUSION: This study demonstrates that NI rates can be significantly reduced by appropriate intervention methods in hospitals that are interested in quality management activities. However, continuous intense efforts are necessary to maintain these improvements. PMID- 11893155 TI - Assessment of and intervention for the misuse of aldehyde disinfectants in Japan. AB - A survey of 145 Japanese hospitals revealed the use of 4 inappropriate aldehyde disinfection methods in 5.5% to 16.8%. Following education, there was discontinuation of 3 of these practices in 26 of 28 affected centers, but only 4 of 18 affected centers stopped the use of formaldehyde vapor cabinets. There is a need to inform hospitals about inappropriate disinfection methods that pose chemical risks to patients and staff. PMID- 11893156 TI - Central venous catheter-associated bloodstream infections in pediatric oncology home care. AB - Fifty-two pediatric oncology patients with central venous catheters (CVCs) who received home care services were studied. Gram-negative organisms were responsible for a greater proportion of CVC-associated bloodstream infections in pediatric oncology patients receiving home care than in hospitalized pediatric oncology patients. PMID- 11893157 TI - Evaluation of mass-transfer coefficient of free fall--cascade-aerator. AB - The mass-transfer coefficient of a free-fall cascade-aerator unit of 15 million litres per day was evaluated for its efficiency in the removal of a class of volatile organics, the trihalomethanes (THMs). These compounds are carcinogenic and occur as a result of chlorination of natural waters. Due to the volatile nature of the THMs, the efficiency of aeration as a potential technique for their removal has been studied. The principle behind aeration is gas-transfer, according to which the gas-liquid interface is hypothesized to consist of a gas and liquid film through which gas is transferred by molecular diffusion until equilibrium is attained. The overall mass transfer coefficient (K(L)) of the aerator considering oxygen as the reference compound, was found to be 29.3 hr(-1) for THMs. PMID- 11893158 TI - Assessment of treatment alternatives for laboratory cod wastewater: a practical comparison with emphasis on cost and performance. AB - This study focused on investigation of treatment alternatives for COD wastewater from academic laboratories, using a number of technologies including chemical reduction/precipitation, ion exchange and adsorption by chitosan. Results showed that high concentrations of 375 mg l(-1) chromium, 1,740 mg l(-1) mercury and 993 mg l(-1) silver in COD wastewater can be reduced to 2.34 mg l(-1), 3.65 mg l(-1) and 1.89 mg l(-1) respectively, by the chemical reduction/precipitation process. Results from ion exchange at a flowrate of 20 ml min(-1) showed breakthrough effluent concentrations obtained at 0.59 mg l(-1) chromium, 3.92 microg l(-1) mercury and 0.65 mg l(-1) silver corresponding to 75.6 l at 63 hr, 40.8 l at 34 hr and 33.6 l at 28 hr respectively. Kinetic and isotherm studies revealed that chitosan can adsorb Cr6+, Hg2+ and Ag+ ions most effectively at a flowrate of 20 ml min(-1) and the optimum pH for feed solution is 4. Chitosan column experiments indicated that average effluent concentrations at breakthrough point for chromium, mercury and silver are 0.76 mg l(-1), 6.04 mg l(-1) and 0.51 mg l(-1) respectively with throughput volumes and retention times of 120 l at 100 hr, 60 l at 50 hr and 48 l at 40 hr. Results of solidification experiments for chemical sludge and residual chitosan based on compressive strength and metal leachability tests showed, that the acceptable ranges of the solidification parameters were: sludge/cement = 0.1-1.0 (weight/weight), water/cement = 0.5-0.6 (weight/weight) and sand/cement = 0.5-3.0 (weight/weight). Operating cost per litre of COD wastewater treated, based on the current prices in Thailand was found to be Baht 19.95 for the chemical reduction/precipitation process, Baht 96.35 for ion exchange treatment and Baht 18.29 for chitosan adsorption. PMID- 11893159 TI - An environmentally sound method for disposal of both ash and sludge wastes by mixing with soil: a case study of Bangkok plain. AB - This study reported the test done on ash-sludge mixture for amendment of soil in pot experiments. Ash-sludge mixture ratio studies revealed that 1:5 fly ash sludge mixture and 1:10 bottom ash-sludge mixture were the optimum mixture ratio that minimized toxic element and provided sufficient nutrients. Experiments indicated that ash-sludge mixtures is more suitable for amendment of acid soil than neutral soil which can increase soil pH and reduce available heavy metal toxicity. The maximum heavy metal adsorption occurred in a pH range of 4 to 6 for all soil studied. The finding also revealed that fly ash application seemed more effective than bottom ash, due to its higher loading rate and metal contents. Heavy metal toxicity was monitored using seed germination test. Marigold and tomato seeds were the two crops selected for this test. Seed germination test result shows that percentage of seed germination increased in pot experiments with sludge only and ash-sludge mixtures. In addition, higher percentages of seed germination were observed to vary with longer incubation time (1-8 weeks). After week 12 of the incubation period, percentage of seed germination began to decline, as a result of reduced soil pH and release of toxic heavy metals. PMID- 11893160 TI - Changes in soil properties of abandoned shrimp ponds in southern Thailand. AB - Chemical soil properties between active shrimp ponds and abandoned ones on the Bangkok soil series were compared, at Ranote District, Songkhla Province in southern Thailand. Soil samples were collected at depth intervals of 0-10, 10-20, 20-30, 30-40 and 40-50 cm from pond bottoms at the same ponds used in a former study conducted in 1994, for a total of 6 ponds with 3 sampling sites for each pond. These ponds were active during the previous study in 1994, abandoned in 1996 and investigated by this study in 1999. All the samples were analyzed for exchangeable Ca, Mg, K and Na electrical conductivity (EC), organic matter, S, P and pH, and statistically compared with the analytical results of the previous study. An increase in amounts of Ca, Mg, K, Na and EC in the abandoned ponds as compared with the active ones by 1.3-3.4, 1.4-2.1, 7.0-30.0, 1.2-6.3 and 1.3-10.9 times respectively was observed. That more of these elements were gained than lost each time the seawater was introduced into the ponds, is explained by the 'Element Input/Output Consideration' as proposed herein. Furthermore, a decrease in organic matter, S and P was also observed in the abandoned pond soils, and attributed to the absence of shrimp food and shrimp excreta following the cessation of shrimp raising activities. An unexpected decline in the soil pH of the abandoned ponds was found as well. Aerobic decomposition of organic matter during the absence of shrimp raising activities caused by soil microorganisms triggering SO2 and H2SO4 formation probably played a more significant role than the increase in the amounts of the basic elements (Ca, Mg, K, and Na) eventually reducing soil pH in the abandoned ponds. The significant depletion of the amounts of organic matter in the abandoned pond soils also supports this observation. PMID- 11893161 TI - Metal levels in raccoon tissues: differences on and off the Department of Energy's Savannah River Site in South Carolina. AB - Levels of arsenic, cadmium, chromium, copper, lead, manganese, selenium, and strontium88 were examined in heart, kidney, muscle, spleen and liver of raccoons (Procyon lotor) from four areas on the Department of Energy's Savannah River Site (SRS), including near a former reactor cooling reservoir and a coal ash basin, and from public hunting areas within 15 km of the site. Mercury is mentioned briefly because it is discussed more fully in another paper. We test the hypotheses that there are no differences in metal levels between raccoons on SRS and off the SRS (off-site), and among different locations on the SRS. Although raccoons collected off-site had significantly lower levels of mercury and selenium in both the liver and kidney, there were few consistencies otherwise. There were significantly higher levels of cadmium in liver of on-site compared to off-site raccoons, and significantly higher levels of chromium and strontium88 in kidney of on-site compared to off-site raccoons. Copper and manganese were highest in the liver; cadmium, lead, mercury and selenium were highest in the liver and kidney; chromium was highest in the spleen and muscle; arsenic was highest in the heart, and strontium88 was slightly higher in the kidney than other organs. Where there were significant differences on site, chromium, manganese were highest in raccoon tissues from Steel Creek; arsenic, lead and selenium were highest in the Ash Basin; cadmium was highest at Upper Three Runs; and strontium88 was highest at Upper Three Runs and Steel Creek. The patterns were far from consistent. PMID- 11893162 TI - Derivation of nutrient guidelines for streams in Victoria, Australia. AB - Human induced increases to nutrient concentrations in streams have led to many agencies developing strategies and criteria for nutrient reduction. National or statewide guidelines are generally inappropriate, due to the natural variability in stream ecosystems within political boundaries. This study used an extant aquatic macroinvertebrate-based regionalisation for the state of Victoria, Australia, as the basis for defining regions of relatively homogeneous environmental character. This enabled the selection of ecologically-based regional reference sites and subsequent characterisation of the nutrient status of these sites. Using an extensive biological and nutrient data base for streams across the State, we calculated 50th and 75th percentile concentrations for reference sites within each region. Using these percentiles in conjunction with 'impact and recovery' studies, we defined nutrient guidelines for each region. Although the nutrient data largely supported the biological regionalisation, patterns in the nutrient data did require some minor modifications for the nutrient regions. Relatively unimpacted regions with reference sites in very good to excellent-condition were assigned guidelines largely based on the 75th percentiles. The more impacted regions, where 'best available' reference sites were of poorer quality, were assigned guidelines based largely on the 50th percentiles. Professional judgement and known extents of impacts across each region provided important contributions to the decision-making process. The derived guideline concentrations are comparable to several cited in the literature and are proposed for use in monitoring, assessment and restoration targets. PMID- 11893163 TI - Transcription factors in cardiogenesis: the combinations that unlock the mysteries of the heart. AB - Heart formation is one of the first signs of organogenesis within the developing embryo and this process is conserved from flies to man. Completing the genetic roadmap of the molecular mechanisms that control the cell specification and differentiation of cells that form the developing heart has been an exciting and fast-moving area of research in the fields of molecular and developmental biology. At the core of these studies is an interest in the transcription factors that are responsible for initiation of a pluripotent cell to become programmed to the cardiac lineage and the subsequent transcription factors that implement the instructions set up by the cells commitment decision. To gain a better understanding of these pathways, cardiac-expressed transcription factors have been identified, cloned, overexpressed, and mutated to try to determine function. Although results vary depending on the gene in question, it is clear that there is a striking evolutionary conservation of the cardiogenic program among species. As we move up the evolutionary ladder toward man, we encounter cases of functional redundancy and combinatorial interactions that reflect the complex networks of gene expression that orchestrate heart development. This review focuses on what is known about the transcription factors implicated in heart formation and the role they play in this intricate genetic program. PMID- 11893164 TI - Tetraspan vesicle membrane proteins: synthesis, subcellular localization, and functional properties. AB - Tetraspan vesicle membrane proteins (TVPs) are characterized by four transmembrane regions and cytoplasmically located end domains. They are ubiquitous and abundant components of vesicles in most, if not all, cells of multicellular organisms. TVP-containing vesicles shuttle between various membranous compartments and are localized in biosynthetic and endocytotic pathways. Based on gene organization and amino acid sequence similarities TVPs can be grouped into three distinct families that are referred to as physins, gyrins, and secretory carrier-associated membrane proteins (SCAMPs). In mammals synaptophysin, synaptoporin, pantophysin, and mitsugumin29 constitute the physins, synaptogyrin 1-4 the gyrins, and SCAMP1-5 the SCAMPs. Members of each family are cell-type-specifically synthesized resulting in unique patterns of TVP coexpression and subcellular colocalization. TVP orthologs have been identified in most multicellular organisms, including diverse animal and plant species, but have not been detected in unicellular organisms. They are subject to protein modification, most notably to phosphorylation, and are part of multimeric complexes. Experimental evidence is reviewed showing that TVPs contribute to vesicle trafficking and membrane morphogenesis. PMID- 11893165 TI - Dynamic changes and the role of the cytoskeleton during the cell cycle in higher plant cells. AB - In higher plant cells microtubules (MTs) show dynamic structural changes during cell cycle progression and play significant roles in cell morphogenesis. The cortical MT (CMT), preprophase band (PPB), and phragmoplast, all of which are plant-specific MT structures, can be observed during interphase, from the late G2 phase to prophase, and from anaphase to telophase, respectively. The CMT controls cell shape, either irreversibly or reversibly, by orientating cellulose microfibril (CMF) deposition in the cell wall; the PPB is involved in determining the site of division; and the phragmoplast forms the cell plate at cytokinesis. The appearance and disappearance of these MT structures during the cell cycle have been extensively studied by immunofluorescence microscopy using highly synchronized tobacco BY-2 cells. Indeed, these studies, together with visualization of MT dynamics in living plant cells using the green fluorescent protein, have revealed much about the modes of MT structural organization, for example, of CMTs at the M/G1 interphase. The microfilaments which also show dynamic changes during the cell cycle, being similar to MTs at particular stages and different at other stages, appear to play roles in supporting MTs. In this article, we summarize our ongoing research and that of related studies of the structure and function of the plant cytoskeleton during cell cycle progression. PMID- 11893166 TI - Sexual dimorphism in the central nervous system of marsupials. AB - It is now evident that gonadal steroids, acting within a limited critical period during fetal or neonatal life, bring about sexual differentiation of both the reproductive tract and the central nervous system (CNS) in eutherians. This results in structural dimorphism in several regions of the brain and spinal cord and the programming of future patterns of adult reproductive behavior. At birth the CNS of marsupials is very underdeveloped and debate continues as to the importance of hormones in its sexual differentiation. Nevertheless, some sexually dimorphic regions have been identified, including the lateral septal nucleus in the hypothalamus and the spinal nucleus of the bulbocavernosus and dorsolateral nucleus in the spinal cord, but interestingly not the cremasteric nucleus, which is dimorphic in eutherians. To date, no apparent sex differences in estrogen and androgen receptor-immunoreactive structures have been detected in the marsupial brain; however, higher levels of aromatase activity during early development in male opossums have been reported. Sex differences have been identified in the localization of cholecystokinin-immunoreactive structures in the marsupial brain indicating that the expression of this neuropeptide is differentially regulated in each sex. A sex difference also exists in the density of arginine vasopressin immunoreactive fibers. Arguments continue as to whether sexually dimorphic behavior in marsupials, as in eutherians, is largely predetermined by hormones acting on the CNS early in development or if it is entirely dependent on the adult steroid hormonal environment. PMID- 11893167 TI - Signaling to gene activation and cell death by tumor necrosis factor receptors and Fas. AB - Tumor necrosis factor (TNF) receptors and Fas elicit a wide range of biological responses, including cell death, cell proliferation, inflammation, and differentiation. The pleiotropic character of these receptors is reflected at the level of signal transduction. The cytotoxic effects of TNF and Fas result from the activation of an apoptotic/necrotic program. On the other hand, TNF receptors, and under certain conditions also Fas, exert a proinflammatory function that results from the induction of several genes. In this context, the transcription factor nuclear factor-kappa B (NF-kappaB) plays an important role. NF-kappaB is also important for the induction of several antiapoptotic genes, which explains at least partially why several cell types can only be killed by TNF in the presence of transcription or translation inhibitors. It is the balance between proapoptotic and antiapoptotic pathways that determines whether a cell will finally die or proliferate. A third signal transduction pathway that is activated in response to TNF is the mitogen-activated protein kinase cascade, which plays an important role in the modulation of transcriptional gene activation. PMID- 11893168 TI - Neuroactive steroid mechanisms and GABA type A receptor subunit assembly in hypothalamic and extrahypothalamic regions. AB - Gonadal- and neuronal-derived steroids are capable of altering brain functions through two basic mechanisms: slow (genomic) and rapid (novel nongenomic membrane) types of activities. The genomic activities that are circumscribed to the numerous neuronal and glial expressed receptor actions involve transcriptional processes regulated largely by classical steroids. On the other hand, rapid nongenomic activities are linked to the stereoselective interactions of potent neuroactive steroids. It appears that both of these steroid mechanisms can be successfully evoked at the ligand-gated heteroligomeric GABA type A receptor. However, the precise structural prerequisites and type of molecular steroid interactions implicated in this neuronal target have not been fully investigated. This article reviews the most common subunits (alpha, beta, and gamma) of the native GABA type A receptor involved in signaling pathways of slow and rapid steroidal mechanisms. Different beta-containing compositions (alpha1beta1-3gamma2) are necessary for the slow type of mechanism, whereas different alpha-containing constructs (alpha2-6beta 1/2 gamma2/2L) are linked to the rapid type. Because of the major role played by neuroactive steroids in GABA dependent neuroendocrine and sociosexual events, distinction of the specific subunit combination is essential not only for elucidating neuronal communicative expressions during such events but also for elucidating their potential neuroprotective role in neurodegenerative disorders. PMID- 11893169 TI - Children of parents with unipolar depression: a comparison of stably remitted, partially remitted, and nonremitted parents and nondepressed controls. AB - This study reports on 122 families with a depressed parent at baseline and matched nondepressed control families. The 10-year course of depression in parents was characterized as stably-, partially-, or non-remitted. At the 10-year follow-up, children of stably-remitted parents had more psychological distress, physical problems, and disturbance than children of controls. Unexpectedly, children of stably-remitted parents had as much distress and disturbance as children of partially- or non-remitted parents. Stably-remitted families emphasized independence less, and organization more, in comparison to controls at 10 years; partially- and non-remitted families were less cohesive and more conflicted than controls. More severe initial or current parental depression was associated with poorer child adaptation, and family functioning explained children's outcomes above and beyond parents' depression. Children living with parents treated for depression are at risk for problems irrespective of the parent's course, perhaps due to poor family functioning. PMID- 11893170 TI - The relationship of adolescent personality and family environment to psychiatric diagnosis. AB - This study examines the relationship of adolescent personality and family environment to psychiatric diagnosis in 170 adolescents admitted to a psychiatric inpatient unit. Patients were administered the Child Assessment Schedule (CAS), the family Environment Scale (FES), and the Millon Adolescent Personality Inventory (MAPI). Adolescent personality and/or family environment were related to 1) major depression, conduct disorder, and attention deficit/hyperactivity disorder in both boys and girls, 2) oppositional defiant disorder, post traumatic stress disorder and overanxious disorder in girls, and 3) dysthymic disorder and alcohol use in boys. The study empirically shows the relationship of both personality and family environment in psychiatric diagnoses. PMID- 11893171 TI - Day length associates with activity level in children living at 60 degrees north. AB - The associations between day length and activity, rest-activity rhythm, and psychiatric symptoms were studied. Sixty-six healthy children participated in the study during one year. They were monitored for 72 consecutive hours with belt worn activity monitors (actigraphs) to obtain objective data on their activity levels during the day and night. In addition, the parents filled out the Child Behavior Checklists. It was found, that the mean total and day and night time activity levels were increased and the relative circadian amplitude blunted with the longer day length. It was concluded that day length was associated with activity level and rest-activity rhythm and this association may reflect the seasonal changes in these parameters. PMID- 11893173 TI - Solution-phase synthesis of aminooxy peptoids in the C to N and N to C directions. AB - [structure: see text] Aminooxy peptoids, which are potential peptidomimetics, were synthesized by a stepwise monomer assembly. Ns-protected N-substituted aminooxyacetate tert-butyl esters were used as a monomer in both the C to N and the N to C directions. Submonomer synthesis of aminooxy peptoids is also described. PMID- 11893174 TI - Novel synthesis of 5,6-dihydro-4H-thieno[3,2-b]pyrrol-5-ones via the rhodium(II) mediated wolff rearrangement of 3-(thieno-2-yl)-3-oxo-2-diazopropanoates. AB - [reaction: see text] Treatment of thioaryolketene S,N-acetals 12 with Hg(OAc)(2) followed by addition of 2-diazo-3-trimethylsilyloxy-3-butenoic acid alkyl esters 15 in CH(2)Cl(2) at room temperature gave 3-(3-alkylamino-5-arylthieno-2-yl)-3 oxo-2-diazopropanoates 16 in good yields. Subsequent reactions of 16 with a catalytic amount of Rh(2)(OAc)(4).2H(2)O in benzene at reflux afforded a mixture of 5,6-dihydro-4H-thieno[3,2-b]pyrrol-5-ones 18 and the corresponding enols 19 in excellent yields. PMID- 11893172 TI - Assessment of anxiety and depression in adolescents with mental retardation. AB - This report examines the concurrent validity of different informant and self report assessment instruments of psychopathology, both general and specific for anxiety and/or depression, in referred mentally retarded adolescents with a depressive and anxiety disorders, according to DSM IV criteria. A consecutive, unselected sample of 50 mildly and moderate mentally retarded adolescents (29 males and 21 females, aged 11.8 to 18 years, mean age 15.1) were assessed using standardized assessment techniques: Psychopathology Instrument for Mentally Retarded Adults (PIMRA) (informant version) (total score, affective and anxiety subscales), Child Behavior Checklist (CBCL) (informant version) (total score, internalizing and externalizing scores, anxiety-depression scale), Zung Self Rating Depression Scale and Zung Self-Rating Anxiety Scale. Patterns of correlation among measures were calculated. PIMRA and CBCL total scores were closely intercorrelated. Internalizing and externalizing scores of CBCL were not intercorrelated, but they both correlated with CBCL and PIMRA total scores. Anxiety measures were positively correlated; they correlated with PIMRA and CBCL total scores, as well as with the internalizing score of CBCL. Depression measures were not correlated; their correlation with more general measures of psychopathology was weak. Clinical implications of these findings are discussed. PMID- 11893175 TI - Synthesis of oligonucleotides containing thiazole and thiazole N-oxide nucleobases. AB - [reaction: see text] The thiazole C-nucleoside analogue was synthesized by the Hantzsch cyclization method to form the thiazole ring and was then converted to the thiazole N-oxide C-nucleoside analogue by peracid oxidation of the heterocycle nitrogen. Incorporation of the thiazole and thiazole N-oxide phosphoramidites into DNA was successful though significant deoxygenation of the N-oxide occurred during DNA assembly. The mechanism proposed for the reduction of the thiazole N-oxide to thiazole involves the formation of an N-oxide phosphite ester. PMID- 11893176 TI - Protons as the triggers to regulate hydrogen-bonding receptors. AB - [reaction: see text] The protonation of alkylamines in two novel receptors results in intramolecular host-guest associations between the resulting ammonium ions and crown ether macrocycles. These interactions result in conformational changes of the receptors and prevent them from acting as hydrogen bond complements for uracil and carboxylate guest species. PMID- 11893177 TI - A formal total synthesis of (+/-)-cephalotaxine using sequential N-acyliminium ion reactions. AB - [reaction: see text] Novel synthesis of cephalotaxine 1 based on tertiary N acyliminium ion chemistry starting from alkynylamide 2 was achieved. The key steps include the preparation of pyrroloisoquinoline 4 from alkynylamide 2, the ring expansion of pyrroloisoquinoline 4 to pyrrolobenzazepine 12, and the construction of cyclopentapyrrolobenzazepine ring system 6, all of which are derived from N-acyliminium ion intermediates. PMID- 11893178 TI - Ruthenium(II) porphyrin catalyzed formation of (Z)-4-alkyloxycarbonyl- methylidene-1,3-dioxolanes from gamma-alkoxy-alpha-diazo-beta-ketoesters. AB - [reaction: see text] Ruthenum(II) porphyrins and dirhodium(II) acetate catalyze cyclization of gamma-alkoxy-alpha-diazo-beta-ketoesters to (Z)-4 (alkyloxycarbonylmethylidene)-1,3-dioxolanes selectively (ca. 68% yield) with no formation of 3(2H)-furanones. Reacting a diazo ketoester with [Ru(II)(TTP)(CO)] [H(2)TTP = meso-tetrakis(p-tolyl) porphyrin] in toluene afforded a ruthenium carbenoid complex, which has been isolated and spectroscopically characterized. A mechanism involving hydrogen atom migration from the C-H bond to the ruthenium carbenoid is proposed. PMID- 11893179 TI - Turn formation initiated by a bissulfoximine motif: synthesis and structural investigation. AB - [reaction: see text] Bissulfoximines containing heterochiral SulfCO-SulfCO units may be used as molecular templates, which induce U-shaped conformations in peptidelike structures. The synthesis of such molecules has been investigated, and the structural requirements of this new turn motif have been identified. PMID- 11893180 TI - A biosynthetic proposal for ring formation in the antitumor agent halichomycin. Asymmetric synthesis of the AB-carbon backbone of halichomycin. AB - [reaction: see text] A biosynthetic proposal for ring formation in the antitumor agent halichomycin is presented in which macrocyclization of the putative prehalichomycin intermediate 1 is the first step. Compound 2 then undergoes dehydration to the alpha-keto N-acylimine 3 followed by tandem nucleophilic addition of the C(16)-hydroxyl to form the hemimacrolactam. A stereospecific Michael ring closure and enol protonation complete C-ring assembly. So far, synthetic efforts toward 1 have resulted in 8. PMID- 11893181 TI - In search of high stereocontrol for the construction of cis-disubstituted cyclopropane compounds. Total synthesis of a cyclopropane-configured urea-PETT analogue that is a HIV-1 reverse transcriptase inhibitor. AB - [reaction: see text] A new azetidine-ligated dirhodium(II) catalyst that possesses a l-menthyl ester attachment provides significant diastereocontrol and high enantiocontrol for the formation of cis-cyclopropane products from reactions of substituted styrenes with diazo esters. PMID- 11893182 TI - Addition of organozinc species to cyclic 1,3-diene monoepoxide. AB - [reaction: see text] The reaction of organozinc reagents (ZnEt(2), ZnPh(2)) with cyclic 1,3-diene monoepoxides in the presence of CF(3)COOH gave the cis-addition products. Lewis acids such as (CF(3)CO(2))(2)Zn and ZnCl(2) mediated the nucleophilic addition of ZnEt(2) to cyclooctadiene monoepoxide with high stereoselectivity. PMID- 11893183 TI - Regioselective synthesis of the bridged tricyclic core of Garcinia natural products via intramolecular aryl acrylate cycloadditions. AB - [reaction: see text] Two different routes to the tricyclic core of Garcinia derived natural products are described. The first approach is based on a tandem Claisen/Diels-Alder rearrangement and delivers the desired lactone 14. The second approach, employing a Wessely oxidation/Diels-Alder protocol, leads to the same caged heterocycle, albeit with modified constitution. PMID- 11893184 TI - Self-assembly of molecular prisms via an organometallic "clip". AB - [reaction: see text] Under the appropriate conditions, the combination of two tritopic pyridyl ligands with three metal-containing molecular "clips" spontaneously generates supramolecular coordination cages with trigonal prismatic frameworks. PMID- 11893185 TI - Photoreduction of benzophenones by amines in room-temperature ionic liquids. AB - [reaction: see text] The amine-mediated photoreduction of benzophenones in room temperature ionic liquids was investigated. Unlike the analogous reaction in organic solvents, the photoreduction produces mainly the corresponding benzhydrol in most cases. Because the reaction consumes only 1 equiv of amine and the solvent can be easily recycled, the photoreduction allows a very clean method for the conversion of benzophenones to benzhydrols. PMID- 11893186 TI - Topological equivalences between organic and coordination polymer crystal structures: an organic ladder formed with three-connected molecular and supramolecular synthons. AB - [structure: see text] Crystal engineering of an organic ladder can be achieved with a T-shaped molecule, 4,4-bis(4'-hydroxyphenyl)-1-cyclohexanol, having three hydroxyl functionalities that can form O-H...O hydrogen-bonded helices. The topology of this network structure finds a parallel in three-connected coordination polymers. PMID- 11893187 TI - Cooperative C(60) binding to a porphyrin tetramer arranged around a p-terphenyl axis in 1:2 host-guest stoichiometry. AB - [structure: see text] Porphyrin tetramer 1 was newly designed and synthesized to construct a novel cooperative [60]fullerene (C(60)) binding system. Compound 1 has a p-terphenyl axis, which is expected to act as a scaffold for a guest binding information transducer. In toluene, 1 can bind 2 equiv of C(60) to produce a 1:2 1/C(60) complex with association constants of 5800 M(-1) (K(1)) and 2000 M(-1) (K(2)). These values are significantly greater than those for control porphyrin dimers such as 2 and 3. PMID- 11893188 TI - Preparation of alpha-sulfenyl enones by thermal fragmentation of beta-sulfenyl enol triflates. AB - [reaction: see text] The synthetic scope and mechanism of the fragmentation of cyclic beta-sulfenyl enol triflates to give alpha-sulfenyl enones are described. This transformation is the central step in a mild, functional group-tolerant method for preparing alpha-sulfenyl enones. PMID- 11893189 TI - Rhodium-catalyzed synthesis of cyclohexenones via a novel [4 + 2] annulation. AB - [reaction: see text] In the presence of a rhodium catalyst, 4-alkynals react with alkynes to furnish cyclohexenones, presumably via metalacycle 1; as far as we are aware, this is the first example of the generation of this class of compounds through such a transformation. In view of the easy accessibility of 4-alkynals (alkynylmetal + alpha,beta-unsaturated aldehyde) and alkynes, this [4 + 2] cyclization constitutes an interesting new approach to functionalized cyclohexenones. PMID- 11893190 TI - Pectenotoxin-2 synthetic studies. 1. Alkoxide precoordination to [Rh(NBD)(DIPHOS 4)]BF(4) allows directed hydrogenation of a 2,3-dihydrofuran-3-ol without competing furan production. AB - [reaction: see text] The alkoxide-directed hydrogenation shown is reported as a key step in a concise synthesis of a fully functionalized precursor to the C29 C40 F/G sector of pectenotoxin-2. PMID- 11893191 TI - Synthesis of marine sponge bisindole alkaloids dihydrohamacanthins. AB - [structure: see text] A convergent synthesis of the marine sponge bisindole alkaloids dihydrohamacanthins is described. The synthesis centers on the construction of 3,5- and 3,6-linked pyrazinones and their reduction to the requisite piperazinones with sodium cyanoborohydride. PMID- 11893192 TI - Synthesis and self-photooxygenation of alkenyl-linked [60]fullerene derivatives. A regioselective ene reaction. AB - [reaction: see text] Alkenyl-linked C(60) derivatives undergo self photooxygenation regioselectively, by the preferential abstraction of allylic hydrogens on the fullerene side of the double bond. PMID- 11893193 TI - A concise, modular synthesis of chiral peraza-macrocycles using chiral aziridines. AB - [reaction: see text] Novel chiral peraza-macrocycles were synthesized from chiral aziridines as a common building block. Efficient syntheses of chiral [26]-N(6), [12]-N(4), [9]-N(3), and [14]-N(4) systems were accomplished. PMID- 11893194 TI - Lewis acid promoted annulation of p-quinoneimines by allylsilanes: a facile entry into benzofused heterocycles. AB - [reaction: see text] Lewis acid promoted addition of allyltriisopropylsilane to p quinoneimines afforded different benzofused heterocycles in moderate to good yields depending on the conditions employed. PMID- 11893196 TI - First fused perpendicular hybrid tetrathiafulvalene (TTF) dimers: a new strategy in pi-extended and rigidified TTF. AB - [structure: see text] The synthesis, theoretical calculations, and crystallographic and electrochemical properties of fused perpendicular tetrathiafulvalene (TTF) dimers incorporating both a TTF unit and a quinonoid pi extended TTF are described as a new strategy for obtaining pi-extended, rigidified, and sulfur-rich analogues of TTF. PMID- 11893195 TI - Mutational analysis of the enterocin favorskii biosynthetic rearrangement. AB - [reaction: see text] A mutational analysis of the enterocin biosynthesis genes revealed that the putative oxygenase and the methyltransferase gene products EncM and EncK, respectively, jointly catalyze a biosynthetic Favorskii-like rearrangement. Inactivation of either gene terminated enterocin production and caused the accumulation of four nonrearranged, nonmethylated polyketides. The structure elucidation of the new wailupemycins E-G is reported. PMID- 11893197 TI - Domino processes as a tool for recovering substandard reactions. Synthesis and use of nitroacetic acid esters and amides. AB - [reaction: see text] Elusive nitroacetic acid esters and amides were obtained through a halogen exchange reaction of the corresponding bromoacetic acid derivatives with polymer-supported nitrite anion. The process is flawed by a side product catalyzed degradation of the products. Domino processes turned out to be a powerful tool for overcoming such drawbacks, converting a substandard reaction into an efficient multicomponent preparation of 4-hydroxy-4,5-dihydroisoxazoles. PMID- 11893198 TI - Asymmetric total synthesis of (+)-fostriecin. AB - [structure: see text] The title compound, a potent protein phosphatase inhibitor and anticancer agent, was prepared by an efficient, multiconvergent asymmetric synthesis. Key transformations include a ring forming olefin metathesis leading to the alpha,beta-unsaturated lactone and creation of the triene moiety via Suzuki cross-coupling. PMID- 11893199 TI - Copper-catalyzed coupling of aryl iodides with aliphatic alcohols. AB - [reaction: see text] A simple and mild method for the coupling of aryl iodides and aliphatic alcohols that does not require the use of alkoxide bases is described. The reactions can be performed in neat alcohol. For more precious alcohols, the etherification was carried out in toluene as solvent using 2 equiv of alcohol. Additionally, the cross-coupling of an optically active benzylic alcohol with an unactivated aryl halide was demonstrated to proceed with complete retention of configuration. PMID- 11893200 TI - A new route to C-aryl glycosides. AB - [reaction: see text] A six-step route for de novo synthesis of C-aryl glycosides based on cycloaddition of an aryl nitrile oxide with 4-pentyn-2-ol has been developed. PMID- 11893202 TI - A new catalytic cross-coupling approach for the synthesis of protected aryl and heteroaryl amidines. AB - [reaction: see text] A new method for the synthesis of protected benzamidines is described. The commercially available 1,3-bis(tert-butoxycarbonyl)-2-methyl-2 thiopseudourea guanidylation reagent, after SEM-protection, functions as an amidine-forming cross-coupling partner under Liebeskind-Srogl conditions. In the presence of copper(I) thiophenecarboxylate (CuTC), the palladium-catalyzed cross coupling of the SEM-protected thiopseudourea reagent with boronic acids affords fully protected benzamidines in good to excellent yield (40-91%). PMID- 11893201 TI - Heteroaromatic thioether-boronic acid cross-coupling under neutral reaction conditions. AB - [reaction: see text] Pi-deficient heteroaromatic thioethers undergo efficient palladium-catalyzed cross-coupling with boronic acids mediated by copper(I) thiophene-2-carboxylate. PMID- 11893203 TI - Regioselective synthesis of bifunctional macrolides for probing ribosomal binding. AB - [structure: see text] Bifunctional macrolides projecting an anchor group to the right or left spatial position of the lactone ring were synthesized. The regioselectivity of the key [3 + 2] cycloaddition process was controlled by the remote cladinose group attached to the C-3 position. These conformationally constrained molecules were employed as molecular probes to study the ribosomal binding sites of bifunctional macrolide antibiotics. PMID- 11893204 TI - Studies toward the synthesis of (-)-zampanolide: preparation of N-acyl hemiaminal model systems. AB - [structure: see text] Synthesis of N-acyl hemiaminal model systems related to the side chain of the antitumor natural product zampanolide is reported. Key steps involve oxidative decarboxylation of N-acyl-alpha-amino acid intermediates, followed by ytterbium triflate mediated solvolysis. Evidence for stabilization of the N-acyl hemiaminal moiety in model compounds by an intramolecular hydrogen bonding network is described. PMID- 11893205 TI - Synthesis, conformational analysis, and bioassay of 9,10-didehydroepothilone D. AB - [reaction: see text] 9,10-Didehydroepothilone D was synthesized, its conformation was studied, and its tubulin polymerization and antiproliferative activity were compared with that of epothilone D and certain analogues. PMID- 11893207 TI - Preparation, properties, and synthetic potentials of novel boronates in a fluorous version (fluorous boronates). AB - [reaction: see text] A series of boronic acids were attached to a fluorous tag by esterification. Functional transformations of these boronates together with the fluorous Suzuki coupling reaction illustrated their usefulness in fluorous-phase techniques. PMID- 11893206 TI - P-chirogenic binaphthyl-substituted monophosphines: synthesis and use in enolate vinylation/arylation reactions. AB - [reaction: see text] New phosphine ligands possessing both axial chirality and a chirogenic phosphorus center were prepared from (R)-2-bromo-2'-N,N (dimethylamino)-1,1'-binaphthyl (1) via a simple Li-halogen exchange protocol. The asymmetric vinylation of a ketone enolate with (R,R(P))-2-(tert butylphenylphosphino)-2'-N,N-(dimethylamino)-1,1'-binaphthyl (2a) afforded the coupling product with good enantiomeric excess. PMID- 11893208 TI - Capture-ROMP-release: application for the synthesis of O-alkylhydroxylamines. AB - [reaction: see text] A new capture-ROMP-release method for chromatography-free purification of N-hydroxysuccinimde Mitsunobu reactions is described. The Mitsunobu reaction captures a variety of alcohols onto a norbornenyl N hydroxysuccinimide monomer. Subjection of the resulting crude reaction mixture to ROM-polymerization generates a polymer that can be precipitated with methanol and filtered from the Mitsunobu byproducts. Treatment of the polymer with hydrazine releases the substrate from the water-soluble polymer, producing a variety of O alkylhydroxylamines with good purity. PMID- 11893209 TI - Radical cascade reaction with 1,4-dienes and 1,4-enynes using 2 (iodomethyl)cyclopropane-1,1-dicarboxylate as a homoallyl radical precursor: one step synthesis of bicyclo[3.3.0]octane derivatives. AB - [reaction: see text] Radical cascade reaction with various 1,4-dienes and 1,4 enynes using dimethyl 2-(iodomethyl)cyclopropane-1,1-dicarboxylate as a homoallyl radical precursor smoothly proceeds through an iodine atom transfer mechanism to give functionalized bicyclo[3.3.0]octane derivatives in good yields. PMID- 11893210 TI - Regio- and diastereoselective reduction of nonenolizable alpha-diketones to acyloins mediated by indium metal. AB - [reaction: see text] Alpha-diketones are efficiently reduced with indium metal in methanol-water in the presence of NH(4)Cl, LiCl, or NaCl to give regio- and diastereoselectively the corresponding acyloins in good to excellent yield. The cleavage of the acyloins under Pb(OAc)(4)/MeOH-PhH condition provides a convenient and regioselective access to highly functionalized cyclopentane carboxaldehydes, potential building blocks in organic syntheses. PMID- 11893211 TI - Synthesis of 2,3-anti-3,4-anti and 2,3-anti-3,4-syn propionate motifs: a diastereoselective tandem sequence of Mukaiyama and free-radical-based hydrogen transfer reactions. AB - [reaction: see text] Reported herein is a strategy employing a Mukaiyama reaction in tandem with a hydrogen transfer reaction for the elaboration of 2,3-anti-3,4 anti and 2,3-anti-3,4-syn propionate motifs. The mode of complexation is controlled through monodentate or chelate pathways for the Mukaiyama reaction to give access to either syn or anti aldol products, precursors of the free-radical reduction reaction. Boron Lewis acid is used to control the free-radical reaction through the exocyclic pathway. PMID- 11893212 TI - Asymmetric total synthesis of (-)-callystatin A employing the SAMP/RAMP hydrazone alkylation methodology. AB - [structure: see text] The asymmetric total synthesis of (-)-callystatin A has been achieved. The key steps generating the stereogenic centers rely on the asymmetric alpha-alkylation of aldehydes or ketones exploiting the SAMP/RAMP hydrazone alkylation methodology, as well as an enzymatic enantioselective reduction of a 3,5-dioxocarboxylate. For the construction of the alkene moieties, highly selective Wittig or Horner-Wadsworth-Emmons reactions were employed. PMID- 11893213 TI - New efficient construction of the ABC core of the taxoids via a sequence of consecutive cobalt(I)-mediated [2 + 2 + 2] and [4 + 2] cyclizations. AB - [reaction: see text] A new and efficient sequence of two consecutive cyclizations, a cobalt(I)-mediated [2 + 2 + 2] cyclotrimerization and a Diels Alder reaction, is proposed to allow the formation of the ABC core of the taxoids in 65% overall yield, starting from an acyclic polyunsaturated precursor. PMID- 11893214 TI - Stereoselective construction of eight-membered carbocycles by brook rearrangement mediated [3 + 4] annulation. AB - [reaction: see text] A newly developed strategy for eight-membered carbocycles via [3 + 4] annulation that involves the combination of beta-substituted acryloylsilanes and enolates of cycloheptenone is described. A unique feature of this annulative approach is its capacity to generate, in two steps, eight membered ring systems containing useful functionalities for further synthetic elaboration from readily available three- and four-carbon components. PMID- 11893215 TI - Polymer-supported O-methylisourea: a new reagent for the O-methylation of carboxylic acids. AB - [reaction: see text] A solid-supported O-methylisourea reagent has been prepared in one step starting from commercially available solid-supported carbodiimide. The isourea reagent has been successfully used for the preparation of methyl esters from the corresponding carboxylic acids. The crude products obtained after resin filtration and solvent evaporation are generally obtained in >98% purity. PMID- 11893216 TI - Mechanochemical synthesis of a novel C(60) dimer connected by a silicon bridge and a single bond. AB - [reaction: see text] A novel C(60) dimer connected by a silicon bridge and a single bond was synthesized by the mechanochemical solid-state reaction employing a high-speed vibration milling technique and fully characterized by the (1)H and (13)C NMR, APCI mass, and UV-vis spectroscopy. The presence of the electronic interaction between the two C(60) cages was demonstrated by the electrochemical method. PMID- 11893218 TI - Chiral 2,2'-bipyridine-type N-monoxides as organocatalysts in the enantioselective allylation of aldehydes with allyltrichlorosilane. AB - [reaction: see text] The Sakurai-Hosomi-type allylation of aromatic and heteroaromatic aldehydes can be catalyzed by the new heterobidenate bipyridine monoxide PINDOX with high enantioselectivities. The sterochemical outcome is mainly controlled by the axial chirality in PINDOX, which in turn is determined by the annulated terpene units. PMID- 11893217 TI - Practical Os/Cu-cocatalyzed air oxidation of allyl and benzyl alcohols at room temperature and atmospheric pressure. AB - [reaction: see text] A new protocol for the oxidation of primary and secondary allyl and benzyl alcohols at room temperature and using 1 atm of air is described. The procedure uses low loadings of copper salts and osmium tetroxide, which is activated with quinuclidine and prereduced with an alkene. Chemoselectivity for allyl and benzyl alcohols is very high, no overoxidation is observed, and the reaction takes place under neutral conditions. PMID- 11893219 TI - Six novel antimycotics. AB - We have reviewed six new antimycotic agents which have potential applications for human cutaneous and mucosal diseases. Information on these six drugs was obtained via an English language search of PubMed through the US National Library of Medicine. The antimycotic agents reviewed include rilopirox, lanoconazole, NND 502, butenafine, eberconazole and voriconazole. Rilopirox is a synthetic pyridone derivative, related to ciclopirox, with a fungicidal action. Rilopirox is a hydrophobic, topical agent with potential application in mucosal candida infections, tinea versicolor and seborrheic dermatitis. Lanoconazole, an imidazole, is a topical agent with potential application in tinea infections and cutaneous candidiasis. The drug has been available for clinical use in Japan since 1994 and once-daily application to affected areas is recommended. In addition to its antifungal effect, animal data suggest that application of lanoconazole 0.5 or 1% cream is associated with accelerated wound healing. NND 502, a stereoselective analog of lanoconazole, is a topical agent with potential application in tinea pedis infection. NND-502 appears to be more effective in inhibiting ergosterol biosynthesis than lanoconazole or bifonazole and clinical trials comparing these agents are awaited. Butenafine is the first member of a new class of antifungals, the benzylamine derivatives, and has been approved for topical use in Japan (since 1992) and the US. Butenafine has a potent fungicidal action and the drug has been shown to be effective in multiple clinical trials in patients with tinea pedis, tinea corporis and tinea cruris. Butenafine has also been reported to exert an anti-inflammatory action after topical application and this may offer potential benefit over other topical antifungal agents. Eberconazole, an imidazole derivative, is a topical antifungal agent that has been shown to be effective in clinical trials in patients with tinea infections. Preliminary data indicate that the eberconazole is effective against some triazole-resistant yeasts such as Candida krusei and Candida glabrata. Voriconazole is an azole antifungal derivative of fluconazole. The drug is available in both oral and parenteral formulations. Oral voriconazole 200mg twice daily has been effective in treating oropharyngeal candidiasis and apergillosis in immunocompromised patients. After 12 weeks' treatment, a similar dosage of the drug elicited a positive response in 69% of nonimmunocompromised patients with invasive aspergillosis. PMID- 11893220 TI - Diagnosis and management of uncommon cutaneous cancers. AB - Uncommon types of cutaneous cancers are mainly cited in the literature as case reports and their etiology, pathogenesis and prognosis have to be surmised because of their rarity. Within this group exist the rare and also the unusual, for instance a relatively common carcinoma arising in a strange circumstance. Initial management of such tumors involves taking a history and performing a thorough examination, allowing a diagnosis to be made. These tend to follow one of three patterns: the lesion is confidently recognized; the lesion is unknown but a likely diagnosis can be made and; the lesion is unknown. It is within the latter two groupings that the uncommon cutaneous cancers exist. A biopsy is then performed to confirm the diagnosis. For large lesions a punch or incisional biopsy is taken which must include a portion of normal skin at the lesion edge. If the lesion is small enough to allow direct closure, an excision biopsy is performed, with a minimum margin of 2mm. Shave biopsy can be employed to confirm a diagnosis, but care must be taken that the subsequent management of the lesion is not adversely affected. With the rare tumors diagnosis can be difficult and there may not be enough tissue in a biopsy for a definitive diagnosis. The whole lesion may therefore need to be excised to obtain a confident diagnosis with further surgical treatment planned as required. Once the diagnosis is established a decision on the method of treatment can be made. The limited literature would suggest that rare skin tumors are unresponsive to radiotherapy and chemotherapy, so the mainstay of treatment is surgery. When there are no established guidelines for the treatment of a particular rare tumor a pathologist can usually provide advice as to the probable nature of the lesion. This allows surgical treatment to be positioned into one of three main groups: lesions that behave like basal cell carcinomas (BCC); lesions that behave like squamous cell carcinomas (SCC) and; lesions that behave like soft tissue sarcomas (STS). It is helpful to have a management plan into which each variety of rare tumor can be fitted, giving guidance as to the best and most appropriate management. Grouping treatment into BCC, SCC or STS-like treatment has been useful. As more becomes known regarding these malignancies their subsequent management can be adjusted accordingly. Currently, it would appear wise to treat each under a broader subgroup. PMID- 11893221 TI - Leishmaniasis: recognition and management with a focus on the immunocompromised patient. AB - Leishmaniasis is a protozoan disease whose clinical manifestations depend both on the infecting species of Leishmania and the immune response of the host. Transmission of the disease occurs by the bite of a sandfly infected with Leishmania parasites. Infection may be restricted to the skin in cutaneous leishmaniasis (CL), to the mucous membranes in mucosal leishmaniasis or spread internally in visceral leishmaniasis (VL). In the last 2 decades, leishmaniasis, especially VL, has been recognized as an opportunistic disease in immunocompromised patients, particularly those infected with HIV. Leishmaniasis is characterized by a spectrum of disease phenotypes that correspond to the strength of the host's cell-mediated immune response. Both susceptible and resistant phenotypes exist within human populations. Clinical cutaneous disease ranges from a few spontaneously-healing lesions, to diffuse external or internal disease, to severe mucous membrane involvement. Spontaneously-healing lesions are associated with positive antigen-specific T cell responsiveness, diffuse cutaneous and visceral disease with T cell non-responsiveness, and mucocutaneous disease with T cell hyperresponsiveness. Current research is focused on determining the extent to which this spectrum of host response is genetically determined. In endemic areas, diagnosis is often made on clinical grounds alone including: small number of lesions; on exposed areas; present for a number of months; resistant to all types of attempted treatments; and usually no pain or itching. Multiple diagnostic techniques are available. When evaluating treatment, the natural history of leishmaniasis must be considered. Lesions of CL heal spontaneously over 1 month to 3 years, while lesions of mucocutaneous and VL rarely, if ever, heal without treatment. Consequently, all the latter patients require treatment. Therapy is not always essential in localized CL, although the majority of such patients are treated. Patients with lesions on the face or other cosmetically important areas are treated to reduce the size of the resultant scar. In addition, the species of parasite should be identified so that infection with Leishmania braziliensis and Leishmania panamensis can be treated to reduce the risk of development of mucocutaneous disease. Treating patients with Leishmania and HIV co-infection requires close monitoring for effectiveness of treatment, especially because of the high relapse rates. Proven treatments include: antimonials, pentamidine, amphotericin B, interferon with antimony. Treatments where current clinical experience is too limited include: allopurinol, ketoconazole, itraconazole, immunotherapy, rifampin, dapsone, localized heat, paromomycin ointment and cryotherapy. Investigational treatments include: WR6026, liposomal amphotericin and miltefosine. In addition, vaccines for leishmaniasis are being investigated in clinical trials. PMID- 11893222 TI - Laser hair removal: guidelines for management. AB - Laser-assisted hair removal is the most efficient method of long-term hair removal currently available. Several hair removal systems have been shown to be effective in this setting: ruby laser (694nm), alexandrite laser (755nm), diode laser (800nm), intense pulsed light source (590 to 1200nm) and the neodymium:yttrium-aluminium-garnet (Nd:YAG) laser (1064nm), with or without the application of carbon suspension. The parameters used with each laser system vary considerably. All these lasers work on the principle of selective photothermolysis, with the melanin in the hair follicles as the chromophobe. Regardless of the type of laser used multiple treatments are necessary to achieve satisfactory results. Hair clearance, after repeated treatments, of 30 to 50% is generally reported 6 months after the last treatment. Patients with dark colored skin (Fitzpatrick IV and V) can be treated effectively with comparable morbidity to those with lighter colored skin. Although there is no obvious advantage of one laser system over another in terms of treatment outcome (except the Nd:YAG laser, which is found to be less efficacious, but more suited to patients with darker colored skin), laser parameters may be important when choosing the ideal laser for a patient. Adverse effects reported after laser-assisted hair removal include erythema and perifollicular edema, which are common, and crusting and vesiculation of treatment site, hypopigmentation and hyperpigmentation (depending on skin color and other factors). Most complications are generally temporary. The occurrence of hypopigmentation after laser irradiation is thought to be related to the suppression of melanogenesis in the epidermis (which is reversible), rather than the destruction of melanocytes. Methods to reduce the incidence of adverse effects include lightening of the skin and sun avoidance prior to laser treatment, cooling of the skin during treatment, and sun avoidance and protection after treatment. Proper patient selection and tailoring of the fluence used to the patient's skin type remain the most important factors in efficacious and well tolerated laser treatment. While it is generally believed that hair follicles are more responsive to treatment while they are in the growing (anagen) phase, conflicting results have also been reported. There is also no consensus on the most favorable treatment sites. PMID- 11893223 TI - Sweet's syndrome: a review of current treatment options. AB - Sweet's syndrome was originally described in 1964 by Dr Robert Douglas Sweet as an 'acute febrile neutrophilic dermatosis'. The syndrome is characterized by pyrexia, elevated neutrophil count, painful red papules, nodules, plaques (which may be recurrent) and an infiltrate consisting predominantly of mature neutrophils that are diffusely distributed in the upper dermis. In addition to skin and mucosal lesions, Sweet's syndrome can also present with extra-cutaneous manifestations. Sweet's syndrome can be classified based upon the clinical setting in which it occurs: classical or idiopathic Sweet's syndrome, malignancy associated Sweet's syndrome and drug-induced Sweet's syndrome. Systemic corticosteroids have been considered the 'gold standard' for the treatment of patients with Sweet's syndrome; in addition, treatment with topical and/or intralesional corticosteroids may be effective as either monotherapy or adjuvant therapy. However, spontaneous resolution of the symptoms and lesions has occurred in several patients with Sweet's syndrome for whom disease-specific therapeutic intervention was not initiated and in some of the patients with drug-induced Sweet's syndrome after withdrawal of the dermatosis-causing medication. Oral therapy with either potassium iodide or colchicine typically results in rapid resolution of Sweet's syndrome symptoms and lesions; therefore, in patients with Sweet's syndrome who have a potential systemic infection or in whom corticosteroids are contraindicated, it is reasonable to initiate treatment with these agents as a first-line therapy. Indomethacin, clofazimine, dapsone, and cyclosporine have also been effective therapeutic agents for managing Sweet's syndrome. However, indomethacin and clofazimine appear less effective than corticosteroids, potassium iodide, and colchicine. Appropriate initial and follow up laboratory monitoring is necessary when treating with either dapsone or cyclosporine because of the potential for severe adverse drug-associated effects. Systemic antibacterials with activity against Staphylococcus aureus frequently result in partial improvement of Sweet's syndrome lesions when they are impetiginized or secondarily infected. In some patients with dermatosis associated bacterial infections, organism-sensitive specific systemic antibacterials have been helpful in the management of their Sweet's syndrome. Although patients with hematologic malignancy-associated Sweet's syndrome often receive cytotoxic chemotherapy agents and antimetabolic drugs for the treatment of their underlying disorder, these agents are seldom used solely for the management of the symptoms and lesions of Sweet's syndrome. The treatment of patients with Sweet's syndrome with either etretinate or interferon-alpha have been reported as single case reports; both patients had improvement of not only their Sweet's syndrome lesions, but also their associated hematologic disorder. PMID- 11893224 TI - Cutaneous drug reaction case reports: from the world literature. AB - Skin disorders are the most common adverse reactions attributed to drugs. Any skin disorder can be imitated, induced or aggravated by drugs. To help you keep up-to-date with the very latest skin reactions occurring with both new and established drugs, this section of the journal brings you information selected from the adverse drug reaction alerting service Reactions Weekly. The following case reports are selected from the very latest to be published in the world dermatology literature. Any claim of a first report has been verified by a search of AdisBase (a proprietary database of Adis International) and Medline. Each case report is assessed for seriousness using the FDA MedWatch definition of serious (patient outcome is: death; life-threatening; hospitalization;disability; congenital anomaly; or requires intervention to prevent permanent impairment or damage). PMID- 11893225 TI - Mechanisms of antimicrobial resistance: their clinical relevance in the new millennium. AB - Antimicrobials show selective toxicity. Suitable targets for antimicrobials to act at include the bacterial cell wall, bacterial protein and folic acid synthesis, nucleic acid metabolism in bacteria and the bacterial cell membrane. Acquired antimicrobial resistance generally can be ascribed to one of five mechanisms. These are production of drug-inactivating enzymes, modification of an existing target, acquisition of a target by-pass system, reduced cell permeability and drug removal from the cell. Introduction of a new antimicrobial into clinical practice is usually followed by the rapid emergence of resistant strains of bacteria in some species that were initially susceptible. This has reduced the long-term therapeutic value of many antimicrobials. It used to be thought that antibacterial resistance was mainly a hospital problem but now it is also a major problem in the community. Organisms in which resistance is a particular problem in the community include members of the Enterobacteriaceae, including Salmonella spp. and Shigella spp., Mycobacterium tuberculosis, Streptococcus pneumoniae, Haemophilus influenzae and Neisseria gonorrhoeae. Multi resistant Gram-negative rods, methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci are major causes of concern in the hospital setting. Prevalence of antibacterial resistance depends both on acquisition and spread. Decreasing inappropriate usage of antimicrobials should lessen the rate of acquisition, and spread can be minimised by sensible infection control measures. PMID- 11893226 TI - Issues of adherence to immunosuppressant therapy after solid-organ transplantation. AB - Nonadherence to immunosuppressant therapy constitutes a major barrier to post transplant care. Failure of transplant recipients to take prescribed drugs properly may not only be a significant obstacle to optimal graft function but it may also result in decreased quality of life and productivity, increased morbidity and healthcare cost, and death. Despite the obvious importance of adherence to immunosuppressant therapy, nonadherence is frequent among transplant recipients, with rates ranging from 2 to 68%. This manuscript briefly discusses several issues concerning adherence to immunosuppressant therapy of solid-organ transplant recipients; presents a literature review concerning adherence to immunosuppressant therapy by solid-organ transplant recipients; and suggests strategies that may be used to enhance medication adherence. Although many of the studies have results that conflict concerning factors associated with immunosuppressive nonadherence, most of the investigators concluded that nonadherent behaviour is usually not predictable. Because of possible adverse events, emphasis should be placed on increasing medication adherence in all transplant recipients. PMID- 11893227 TI - Management of fibromyalgia: what are the best treatment choices? AB - Fibromyalgia still represents an enigma to modern medicine and the aetiopathogenesis is far from explored. The management of patients with fibromyalgia is thus mostly based on empirical research, and only a few controlled studies have been performed. Basic drug therapy rests on the administration of amitriptyline and conventional analgesics. Such therapy should be initiated only after careful patient information and delineation of therapeutic goals are provided. Any drug therapy should be administered in combination with physical treatment and cognitive behavioural therapy. Because of the appearing contours of pathogenic mechanisms, hopefully a number of new drugs will be available to the patients with this complex pain syndrome in the near future. PMID- 11893228 TI - Effect of antiepileptic drugs on cognitive function in individuals with epilepsy: a comparative review of newer versus older agents. AB - Several 'new' antiepileptic drugs (AEDs), i.e. oxcarbazepine, vigabatrin, lamotrigine, zonisamide, gabapentin, tiagabine, topiramate and levetiracetam have been introduced into clinical practice within the last decade. Most of these new drugs are at least as effective as the 'old' AEDs [phenytoin, phenobarbital (phenobarbitone), valproic acid (sodium valproate) and carbamazepine] and, in general, they seem to be better tolerated than the old drugs. The new AEDs might have less influence on cognitive functions but the aspect has not been systematically studied. Neuropsychological testing has been the major method of objectively examining cognitive function related to the use of AEDs but a number of methodological problems blur the results. Alteration of cognition might reflect a chronic adverse effect of AEDs but the negative effects of the drugs are only one of several factors that may influence cognition. In addition, subjective complaints about cognitive deficits (e.g. memory problems or attention) may also reflect other aspects of adverse effects than those concerning specific cognitive functions (e.g. mood and anxiety). This review focuses on studies of the cognitive effects of the new AEDs, and in particular on studies that compare cognitive effects of the old and new drugs. In general, the new AEDs seem to display no or minor negative cognitive effects. In studies in which new AEDs have been compared with old AEDs, there was a tendency in favour of the new AEDs in some of the studies. PMID- 11893230 TI - Drotrecogin alfa (activated). AB - Drotrecogin alfa (activated), recombinant human activated protein C, inhibits coagulation and inflammation and promotes fibrinolysis in patients with severe sepsis. 850 patients with severe sepsis treated with intravenous drotrecogin alfa (activated) 24 microg/kg/h for 96 hours had a significantly greater reduction in 28-day all-cause mortality (24.7%) than 840 placebo recipients (30.8%) in a randomised, double-blind, placebo-controlled study. The drug was associated with a 19.4% reduction in the relative risk of death at 28 days compared with placebo. Baseline characteristics of and pre-existing conditions in patients with sepsis appeared to have no effect on the efficacy of drotrecogin alfa (activated). A significantly greater reduction in median percentage change from baseline plasma D-dimer levels (a coagulation marker) was seen with drotrecogin alfa (activated) treatment than with placebo on study days 1 to 7 in patients with severe sepsis. On study days 1, 4, 5, 6 and 7, a significantly greater median reduction in interleukin-6 levels (an inflammation marker) from baseline was seen with drotrecogin alfa (activated) treatment than placebo. Drotrecogin alfa (activated) was associated with an increased incidence of serious bleeding events during the infusion period [2.4% vs 1.0% with placebo; p = 0.024] and the 28-day study period (3.5 vs 2.0%; p = 0.06) of the efficacy trial. This increase was primarily related to procedure-related events; there were no significant differences between the treatment groups in nonprocedure-related serious bleeding events. The most frequent site of bleeding was the gastrointestinal tract. With the exception of bleeding events, there were no clinically significant differences between treatment groups in the efficacy trial in the incidence of adverse events. Of the 210 deaths in patients with severe sepsis treated with drotrecogin alfa (activated) 24 microg/kg/h in the efficacy trial, four deaths due to haemorrhage and one due to cerebral oedema were possibly related to the study drug. PMID- 11893229 TI - Drugs affecting homocysteine metabolism: impact on cardiovascular risk. AB - Elevated total plasma homocysteine has been established as an independent risk factor for thrombosis and cardiovascular disease. A strong relationship between plasma homocysteine levels and mortality has been reported in patients with angiographically confirmed coronary artery disease. Homocysteine is a thiol containing amino acid. It can be metabolised by different pathways, requiring various enzymes such as cystathionine beta-synthase and methylenetetrahydrofolate reductase. These reactions also require several co-factors such as vitamin B6 and folate. Medications may interfere with these pathways leading to an alteration of plasma homocysteine levels. Several drugs have been shown to effect homocysteine levels. Some drugs frequently used in patients at risk of cardiovascular disease, such as the fibric acid derivatives used in certain dyslipidaemias and metformin in type 2 (non-insulin-dependent) diabetes mellitus, also raise plasma homocysteine levels. This elevation poses a theoretical risk of negating some of the benefits of these drugs. The mechanisms by which drugs alter plasma homocysteine levels vary. Drugs such as cholestyramine and metformin interfere with vitamin absorption from the gut. Interference with folate and homocysteine metabolism by methotrexate, nicotinic acid (niacin) and fibric acid derivatives, may lead to increased plasma homocysteine levels. Treatment with folate or vitamins B6 and B12 lowers plasma homocysteine levels effectively and is relatively inexpensive. Although it still remains to be demonstrated that lowering plasma homocysteine levels reduces cardiovascular morbidity, surrogate markers for cardiovascular disease have been shown to improve with treatment of hyperhomocystenaemia. Would drugs like metformin, fibric acid derivatives and nicotinic acid be more effective in lowering cardiovascular morbidity and mortality, if the accompanying hyperhomocysteinaemia is treated? The purpose of this review is to highlight the importance of homocysteine as a risk factor, and examine the role and implications of drug induced modulation of homocysteine metabolism. PMID- 11893233 TI - Alfuzosin: a review of the therapeutic use of the prolonged-release formulation given once daily in the management of benign prostatic hyperplasia. AB - Alfuzosin, a quinazoline derivative, is a selective and competitive alpha(1) adrenoceptor antagonist. It distributes preferentially in the prostate, compared with plasma, and decreases the sympathetically controlled tone of prostatic smooth muscle. As a result lower urinary tract symptoms suggestive of benign prostatic hyperplasia (BPH) are improved. The once-daily formulation of alfuzosin contains inactive barrier layers which have been added to the planar surfaces of compressed tablets. Drug release is sustained over 20 hours with a near constant dissolution rate between 2 and 12 hours. Mean values for area under the plasma concentration-time curve over 24 hours (AUC(24)) were similar after administration of prolonged-release alfuzosin 10mg once daily and immediate release alfuzosin 2.5mg three times daily. Likewise, similar AUC(24) values were reported when prolonged-release alfuzosin 10mg once daily and sustained-release alfuzosin 5mg twice daily were compared. These data suggest that these alfuzosin regimens provide similar average systemic exposure. Data from short- (3 months) and long-term (up to 12 months) clinical trials show that the prolonged-release formulation of alfuzosin controls the symptoms associated with BPH as effectively as immediate-release alfuzosin 2.5mg three times daily and clinical improvement is maintained for up to 1 year. Improvements in International Prostate Symptom Score, maximum urinary flow rate and quality-of-life index were improved to a similar extent in patients treated with immediate- or prolonged-release alfuzosin and improvements were statistically significant compared with placebo. Prolonged release alfuzosin 10mg is well tolerated and the overall incidence of adverse events is similar to that seen with placebo. The once-daily formulation of alfuzosin 10mg caused fewer vasodilatory adverse events than immediate-release alfuzosin 2.5mg three times daily and caused only slight decreases in systolic and diastolic blood pressure which were not clinically significant and did not differ significantly from those with placebo. No dosage titration is required. The incidence of ejaculatory disorders was <1%. CONCLUSION: Prolonged-release alfuzosin 10mg once daily controls symptoms associated with BPH throughout a 24 hour dosage interval as effectively as immediate-release alfuzosin 2.5mg three times daily but with fewer vasodilatory adverse events. A nonblind extension study showed that clinical benefits were maintained for up to 1 year and the once daily 10mg formulation continued to be well tolerated, particularly in terms of cardiovascular effects and sexual function. Thus, for the medical management of men with BPH, prolonged-release alfuzosin 10mg is an effective, well tolerated and convenient treatment option. PMID- 11893235 TI - Botulinum toxin B: a review of its therapeutic potential in the management of cervical dystonia. AB - Botulinum toxins are well known as the causative agents of human botulism food poisoning. However, in the past two decades they have become an important therapeutic mainstay in the treatment of dystonias including cervical dystonia, a neurological disorder characterised by involuntary contractions of the cervical and/or shoulder muscles. The toxins inhibit acetylcholine release from neuromuscular junctions, producing muscle weakness when injected into dystonic muscles. Data from three double-blind, randomised, placebo-controlled trials demonstrate that botulinum toxin B effectively reduces the severity, disability and pain of cervical dystonia. In two of the trials, mean Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS)-Total score at week 4 (primary efficacy measure) after botulinum toxin B 10 000U was reduced by 11.7 (25%) or 11 (21%) compared with baseline. These changes were significantly greater than those obtained with placebo [4.3 (10%) or 2 (4%)] and were generally similar in patients who were responsive or resistant to botulinum toxin A. Statistically significant benefits compared with placebo were also evident for a range of other efficacy parameters including TWSTRS-Severity, -Pain and -Disability subscales, patient- assessed pain and patient-/physician-assessed global improvement ratings. In another trial, the percentage of patients with botulinum toxin A resistant or -responsive cervical dystonia who had a > or =20% improvement in the TWSTRS-Total score between baseline and week 4 was significantly higher with botulinum toxin B 2500 to 10 000U (58 to 77%) than with placebo (27%). Overall, botulinum toxin B was generally well tolerated. The most frequently reported treatment-related adverse events were dry mouth and dysphagia. Most adverse events in patients receiving botulinum toxin B were mild or moderate; no serious adverse events or laboratory abnormalities were associated with the use of botulinum toxin B and, where reported, no patients discontinued from any of the clinical trials as a result of adverse events. CONCLUSIONS: Botulinum toxin B has shown clinical efficacy in patients with cervical dystonia at doses up to 10 000U and is generally well tolerated. Its efficacy extends to patients who are resistant to botulinum toxin A. Although the potential for secondary resistance to botulinum toxin B remains unclear, it may occur less than with botulinum toxin A because methods for manufacturing commercially available botulinum toxin B do not include lyophilisation and the product does not require reconstitution before use. As injection with botulinum toxin is generally considered the treatment of choice for patients with cervical dystonia, botulinum toxin B should be considered a potential treatment option in this setting. PMID- 11893237 TI - Initiation of apoptosis in cells exposed to medium from the progeny of irradiated cells: a possible mechanism for bystander-induced genomic instability? AB - Genomic instability and bystander effects have recently been linked experimentally both in vivo and in vitro. The aim of the present study was to determine if medium from irradiated cells several passages distant from the original exposure could initiate apoptosis in unirradiated cells. Human keratinocytes (from the HPV-G cell line) were irradiated with 0.5 Gy or 5 Gy gamma rays. Medium was harvested at each passage up to the 7th passage (approximately 35 population doublings) postirradiation and transferred to unirradiated keratinocytes. Intracellular calcium levels, mitochondrial membrane potential, and the level of reactive oxygen species were all monitored for 24 h after medium transfer. Rapid calcium fluxes (within 30 s), loss of mitochondrial membrane potential, and increases in reactive oxygen species (from 6 h after medium transfer) were observed in the recipient cells. There was no significant difference between medium conditioned by cells irradiated with 0.5 or 5 Gy. The effect of medium from progeny was the same as the initial effect reported previously and did not diminish with increasing passage number. The data suggest that initiating events in the cascade that leads to apoptosis are induced in unirradiated cells by a signal produced by irradiated cells and that this signal can still be produced by the progeny of irradiated cells for several generations. PMID- 11893234 TI - Paroxetine: an update of its use in psychiatric disorders in adults. AB - Paroxetine is a selective serotonin reuptake inhibitor (SSRI), with antidepressant and anxiolytic activity. In 6- to 24-week well designed trials, oral paroxetine 10 to 50 mg/day was significantly more effective than placebo, at least as effective as tricyclic antidepressants (TCAs) and as effective as other SSRIs and other antidepressants in the treatment of major depressive disorder. Relapse or recurrence over 1 year after the initial response was significantly lower with paroxetine 10 to 50 mg/day than with placebo and similar to that with imipramine 50 to 275 mg/day. The efficacy of paroxetine 10 to 40 mg/day was similar to that of TCAs and fluoxetine 20 to 60 mg/day in 6- to 12-week trials in patients aged > or =60 years with major depression. Paroxetine 10 to 40 mg/day improved depressive symptoms to an extent similar to that of TCAs in patients with comorbid illness, and was more effective than placebo in the treatment of dysthymia and minor depression. Paroxetine 20 to 60 mg/day was more effective than placebo after 8 to 12 weeks' treatment of obsessive-compulsive disorder (OCD), panic disorder, social anxiety disorder (social phobia), generalised anxiety disorder (GAD) and post-traumatic stress disorder (PTSD). Improvement was maintained or relapse was prevented for 24 weeks to 1 year in patients with OCD, panic disorder, social anxiety disorder or GAD. The efficacy of paroxetine was similar to that of other SSRIs in patients with OCD and panic disorder and similar to that of imipramine but greater than that of 2'chlordesmethyldiazepam in patients with GAD. Paroxetine is generally well tolerated in adults, elderly individuals and patients with comorbid illness, with a tolerability profile similar to that of other SSRIs. The most common adverse events with paroxetine were nausea, sexual dysfunction, somnolence, asthenia, headache, constipation, dizziness, sweating, tremor and decreased appetite. In conclusion, paroxetine, in common with other SSRIs, is generally better tolerated than TCAs and is a first line treatment option for major depressive disorder, dysthymia or minor depression. Like other SSRIs, paroxetine is also an appropriate first-line therapy for OCD, panic disorder, social anxiety disorder, GAD and PTSD. Notably, paroxetine is the only SSRI currently approved for the treatment of social anxiety disorder and GAD, which makes it the only drug of its class indicated for all five anxiety disorders in addition to major depressive disorder. Thus, given the high degree of psychiatric comorbidity of depression and anxiety, paroxetine is an important first-line option for the treatment of major depressive disorder, OCD, panic disorder, social anxiety disorder, GAD and PTSD. PMID- 11893236 TI - The radiation-induced bystander effect for clonogenic survival. AB - It has long been accepted that the radiation-induced heritable effects in mammalian cells are the result of direct DNA damage. Recent evidence, however, suggests that when a cell population is exposed to a low dose of alpha particles, biological effects occur in a larger proportion of cells than are estimated to have been traversed by alpha particles. Experiments involving the Columbia University microbeam, which allows a known fraction of cells to be traversed by a defined number of alpha particles, have demonstrated a bystander effect for clonogenic survival and oncogenic transformation in C3H 10T(1/2) cells. When 1 to 16 alpha particles were passed through the nuclei of 10% of a C3H 10T(1/2) cell population, more cells were unable to form colonies than were actually traversed by alpha particles. Both hit and non-hit cells contributed to the outcome of the experiments. The present work was undertaken to assess the bystander effect of radiation in only non-hit cells. For this purpose, Chinese hamster V79 cells transfected with hygromycin- or neomycin-resistance genes were used. V79 cells stably transfected with a hygromycin resistance gene and stained with a nuclear dye were irradiated with the charged-particle microbeam in the presence of neomycin-resistant cells. The biological effect was studied in the neomycin resistant V79 cells after selective removal of the hit cells with geneticin treatment. PMID- 11893238 TI - Induction of adaptive response by low-dose radiation in RIF cells transfected with Hspb1 (Hsp25) or inducible Hspa (Hsp70). AB - An adaptive response results in a reduced effect of a high challenging dose of a stressor after a smaller, inducing dose has been applied a few hours earlier. Radiation-induced fibrosarcoma (RIF) cells did not show an adaptive response, i.e. a reduced effect from a high challenging dose (2 Gy) of a radiation after a priming dose (1 cGy) had been applied 4 or 7 h earlier, but cells of a thermoresistant clone (TR) derived from RIF cells did. Since the expression of inducible Hspa (also known as Hsp70) and Hspb1 (also known as Hsp25) was different in these two cell lines, the role of inducible Hspa and Hspb1 in the adaptive response was examined. When RIF cells were transfected with inducible Hspa or Hspb1, both radioresistance measured by clonogenic assays and a reduction of apoptosis were detected. The adaptive response was also acquired by these two cell lines. The inducible Hspa transfectant showed a more pronounced adaptive response than the Hspb1 transfectant. Based on these results, it appears that inducible Hspa and Hspb1 are at least partly responsible for the induction of the adaptive response in these cells. Moreover, when inducible Hspa or Hspb1 was transfected into RIF cells, co-regulation of the two genes was detected. Heat shock factor (Hsf) was found to be at least partially responsible for the induction of the adaptive response in these cells. PMID- 11893239 TI - Comet assay measurements of DNA damage in cells by laser microbeams and trapping beams with wavelengths spanning a range of 308 nm to 1064 nm. AB - DNA damage induced in NC37 lymphoblasts by optical tweezers with a continuous wave Ti:sapphire laser and a continuous-wave Nd:YAG laser (60-240 mW; 10-50 TJ/m2; 30-120 s irradiation) was studied with the comet assay, a single-cell technique used to detect DNA fragmentation in genomes. Over the wavelength range of 750-1064 nm, the amount of damage in DNA peaks at around 760 nm, with the fraction of DNA damage within the range of 750-780 nm being a factor of two larger than the fraction of DNA damage within the range of 800-1064 nm. The variation in DNA damage was not significant over the range of 800-1064 nm. When the logarithm of damage thresholds measured in the present work, as well as values reported previously in the UV range, was plotted as a function of wavelength, a dramatic wavelength dependence became apparent. The damage threshold values can be fitted on two straight lines, one for continuous-wave sources and the other for pulsed sources, irrespective of the type of source used (e.g. classical lamp or laser). The damage threshold around 760 nm falls on the line extrapolated from values for UV-radiation-induced damage, while the data for 800-1064 nm fall on a line that has a different slope. The change in the slope between 320 and 340 nm observed earlier is consistent with a well-known change in DNA-damaging mechanisms. The change observed around 780 nm is therefore suggestive of a further change in the mechanism(s). The data from this work together with our previous measurements provide, to the best of our knowledge, the most comprehensive view available of the DNA damage produced by microfocused light. PMID- 11893240 TI - A critical stage in spermatogenesis for radiation-induced cell death in the medaka fish, Oryzias latipes. AB - To ensure the high-fidelity transmission by reproductive cells of genetic information from generation to generation, cells have evolved surveillance systems to eliminate genomic lesions by inducing cell suicide and/or DNA repair. In this report, gamma-ray-induced cell death was investigated using the medaka fish, Oryzias latipes, because of the ease with which the differentiation stages of its spermatogenic cells can be identified. After 4.75 Gy gamma irradiation, the maximum rate of death of spermatogonial stem cells was observed at 18 h, and that of differentiating spermatogonia was at 12 h, followed by a peak in the extent of DNA fragmentation detected by the TUNEL assay. Dose-response curves for the death rate showed an obvious increase in the death rate for early differentiating spermatogonia even after 0.11 Gy irradiation, whereas there were no such increases for spermatogonial stem cells and late-differentiating spermatogonia. In the male germ cells of this fish, the stage during spermatogenesis most sensitive to radiation-induced cell death is in early differentiating spermatogonia, the immediate descendants of the stem cells. These spermatogonia may have a rigorous surveillance system for genomic lesions induced in spermatogonial stem cells. PMID- 11893241 TI - Angiotensin II blockade reduces radiation-induced proliferation in experimental radiation nephropathy. AB - Total-body irradiation or renal irradiation is followed by a well-defined sequence of changes in renal function leading eventually to renal failure. Previous studies in a rat model have shown that inhibition of angiotensin converting enzyme or blockade of angiotensin II receptors can prevent the structural and functional changes that occur after renal irradiation, and that these interventions are particularly important between 3 and 10 weeks after irradiation. We have now shown that in the same rat model, total-body irradiation induces proliferation of renal tubular cells (i.e., an increase in the number of cells staining positive for proliferating cell nuclear antigen) within 5 weeks after irradiation. Treatment with an angiotensin II receptor blocker delays this radiation-induced tubular proliferation and decreases its magnitude. Renal radiation also induces proliferation of glomerular cells, but the relative increase in glomerular proliferation is not as great as that seen in renal tubular cells, and the increase is not delayed or decreased by treatment with an angiotensin II receptor blocker. We hypothesize that angiotensin II receptor blockers exert their beneficial effect in radiation nephropathy by delaying the proliferation (and hence the eventual mitotic death) of renal tubular cells that have been genetically crippled by radiation. PMID- 11893242 TI - Topical antioxidant vitamins C and E prevent UVB-radiation-induced peroxidation of eicosapentaenoic acid in pig skin. AB - Eicosapentaenoic acid protects against UV-radiation-induced immunosuppression and photocarcinogenesis, but it is also prone to oxidative degradation, which may reduce or abolish its beneficial effects. The protective effect of topically applied vitamin E, vitamin C, or both against UVB-radiation-induced lipid peroxidation in the presence of eicosapentaenoic acid was investigated using an ex vivo pig skin model. Changes in the bioavailability of both antioxidants induced by UV radiation were studied in different skin compartments. The UVB radiation dose used (25 kJ/m2) was similar to that required to induce immunosuppression in BALB/c mice. Exposure of pig skin with an epidermal eicosapentaenoic acid content of 1.0 +/- 0.3 mol% to UVB radiation resulted in an 85% increase of epidermal lipid peroxidation (P < 0.005). Topical application of vitamin E or vitamin C 60 min prior to UVB irradiation resulted in a major increase in both antioxidants in the stratum corneum and viable epidermis (P < 0.05). Vitamin E and vitamin C completely protected against UVB-radiation-induced lipid peroxidation (P < 0.005), but compared to vitamin E, a 500-fold higher vitamin C dose was needed. UVB irradiation induced a vitamin E consumption of up to 100% in the stratum corneum and viable epidermis, and a vitamin C consumption of only 21% in the stratum corneum. Simultaneously applied vitamin E and vitamin C also completely protected against UVB-radiation-induced lipid peroxidation (P < 0.05), and lower antioxidant doses were needed compared to vitamin E or vitamin C alone. In the presence of vitamin C, epidermal vitamin E was more stable upon UVB irradiation (P < 0.05), suggesting interaction between vitamin E and vitamin C. In conclusion, topically applied vitamin E and/or vitamin C efficiently protect against UVB-radiation-induced lipid peroxidation in the presence of eicosapentaenoic acid. The beneficial biological effects of eicosapentaenoic acid may therefore be improved if vitamin E and/or vitamin C are present in sufficient amounts. The ex vivo pig skin model provides a useful tool for assessing short term biochemical effects related to UVB radiation, without the use of living experimental animals. PMID- 11893243 TI - Skin cancer after X-ray treatment for scalp ringworm. AB - Some 2,224 children given X-ray therapy for tinea capitis (ringworm of the scalp) have been followed for up to 50 years to determine cancer incidence, along with a control group of 1,380 tinea capitis patients given only topical medications. The study found a relative risk (RR) of 3.6 (95% confidence interval, 2.3-5.9) for basal cell skin cancer (BCC) of the head and neck among irradiated Caucasians (124 irradiated cases and 21 control cases), in response to a scalp dose of about 4.8 Gy. No melanomas of the head and neck have been seen, and only a few squamous cell carcinomas. About 40% of irradiated cases have had multiple BCCs, for a total of 328 BCCs. Although 25% of both the irradiated and control groups are African-American, only 3 skin cancers have been seen among them, all in the irradiated group, indicating the importance of susceptibility to UV radiation as a cofactor. Light complexion, severe sunburning and North European ancestry were predictive of BCC risk in the irradiated group, but chronic sun exposure was not. Children irradiated at young ages had the highest BCC risk. The RR for BCC risk is approximately constant with time since exposure, suggesting that risk will probably last for a lifetime. PMID- 11893245 TI - Bone tumorigenesis induced by alpha-particle radiation: mapping of genetic loci influencing predisposition in mice. AB - The present study was carried out to determine the extent to which genetic factors modify the incidence of radiation-induced bone tumorigenesis in mice, and to map putative susceptibility genes. We conducted a genome-wide linkage analysis in a cohort of 47 interstrain backcrossed mice. After the mice were injected with the bone-seeking alpha-particle-emitting radionuclide (227)Th, 21 of the mice developed osteosarcomas. Two loci, one on chromosome 7 close to D7Mit145 and a second on chromosome 14 (D14Mit125), exhibited suggestive linkage to osteosarcoma predisposition, with LOD scores of 1.37 and 1.05, respectively. The LOD score increased considerably when interaction between these two loci was taken into account (LOD = 3.48). Nine of 12 mice inheriting a susceptibility allele at both loci developed osteosarcomas after (227)Th injection, compared to only four osteosarcomas in 18 animals that did not inherit either of the susceptibility alleles. Variance component analysis revealed that these genetic factors determine approximately one-fifth of the total incidence of osteosarcomas. This study demonstrates the presence of a genetic component that modulates predisposition to radiation-induced osteosarcoma. PMID- 11893244 TI - Cancer incidence among Swedish patients exposed to radioactive thorotrast: a forty-year follow-up survey. AB - Thorotrast is an alpha-particle-emitting radiological contrast medium that caused chronic exposure to internal alpha-particle radiation when it was administered systemically. Cancer incidence in 432 Swedish patients exposed to Thorotrast was evaluated by computerized linkage of the cohort with the Swedish Cancer Register. Standardized incidence ratios (SIRs) were calculated as the ratio of observed cases in the cohort to expected cases in the general population. A total of 170 cancers occurring in 152 individuals were reported, whereas only 57 cases were expected. The SIR was significantly increased for cancer at all sites (3.0), with the largest excesses noted for primary liver and gallbladder cancer (SIR = 39.2). Other significantly elevated risks were observed for liver cancer not specified as primary, small intestine cancer, stomach cancer, leukemia, kidney cancer, CNS tumors, and pancreatic cancer. Among women, there was a significantly increased risk for lung cancer, based on a small number. Our results show that cumulative radiation exposure is directly related to carcinogenesis in the liver and gallbladder, which is consistent with earlier findings. In addition, there may be a relationship between radiation exposure and the development of other solid tumors. PMID- 11893246 TI - The response of tissue-equivalent proportional counters to heavy ions. AB - The paper presents a theoretical model for the response of a tissue-equivalent proportional counter (TEPC) irradiated with charged particles. Heavy ions and iron ions in particular constitute a significant part of radiation in space. TEPCs are used for all space shuttle and International Space Station (ISS) missions to estimate the dose and radiation quality (in terms of lineal energy) inside spacecraft. The response of the tissue-equivalent proportional counters shows distortions at the wall/cavity interface. In this paper, we present microdosimetric investigation using Monte Carlo track structure calculations to simulate the response of a TEPC to charged particles of various LET (1 MeV protons, 2.4 MeV alpha particles, 46 MeV/nucleon 20Ne, 55 MeV/nucleon 20Ne, 45 MeV/nucleon 40Ar, and 1.05 GeV/nucleon 56Fe). Data are presented for energy lost and energy absorbed in the counter cavity and wall. The model calculations are in good agreement with the results of Rademacher et al. (Radiat. Res. 149, 387-389, 1998), including the study of the interface between the wall and the sensitive region of the counter. It is shown that the anomalous response observed at large event sizes in the experiment is due to an enhanced entry of secondary electrons from the wall into the gas cavity. PMID- 11893247 TI - Involvement of TP53 in apoptosis induced in human lymphoblastoid cells by fast neutrons. AB - We investigated the involvement of TP53 in apoptosis induced by fast neutrons in cells of three human B-lymphoblast cell lines derived from the same donor and differing in TP53 status: TK6 (wild-type TP53), WTK1 (mutant TP53) and NH32 (knockout TP53). Cells were exposed to X rays or to fast neutrons at doses ranging from 0.5 to 8 Gy. Apoptosis was determined by measurements of the sub-G0 /G1-phase DNA content and by the externalization of phosphatidylserine. Fast neutrons induced extensive apoptosis in TK6 cells, as shown by the formation of hypodiploid particles, the externalization of phosphatidylserine, and the activation of caspases. In contrast, cell death was triggered at a significantly lower rate in cells lacking functional TP53. However, TP53-independent cell death also expressed the morphological and biochemical hallmarks of apoptosis. Proliferation tests and clonogenic assays showed that fast neutrons can nevertheless kill WTK1 and NH32 cells efficiently. The absence of functional TP53 only delays radiation-induced cell death, which is also mediated by caspases. These results indicate that fast-neutron irradiation activates two pathways to apoptosis and that the greater relative biological effectiveness of fast neutrons reflects mainly an increase in clonogenic cell death. PMID- 11893248 TI - Relative biological effectiveness of 144 keV neutrons in producing dicentric chromosomes in human lymphocytes compared with 60Co gamma rays under head-to-head conditions. AB - The RBE for neutrons was assessed in a head-to-head experiment in which cultures of lymphocytes from the same male donor were irradiated simultaneously with 144 keV neutrons and with 60Co gamma rays as the reference radiation and evaluated using matched time, culture conditions, and the end point of chromosomal aberrations to avoid potential confounding factors that would influence the outcome of the experiment. In addition, the irradiation time was held constant at 2 h for the high-dose groups for both radiation types, which resulted in rather low dose rates. For the induction of dicentric chromosomes, the exposure to the 144 keV neutrons was found to be almost equally as effective (yield coefficient alpha(dic) = 0.786 +/- 0.066 dicentrics per cell per gray) as that found previously for irradiation with monoenergetic neutrons at 565 keV (alpha(dic) = 0.813 +/- 0.052 dicentrics per cell per gray) under comparable exposure and culture conditions (Radiat. Res. 154, 307-312, 2000). However, the values of the maximum low-dose RBE (RBE(m)) relative to 60Co gamma rays that were determined in the present and previous studies show an insignificant but conspicuous difference: 57.0 +/- 18.8 and 76.0 +/- 29.5, respectively. This difference is mainly due to the difference in the alpha(dic) value of the 60Co gamma rays, the reference radiation, which was 0.0138 +/- 0.0044 Gy(-1) in the present study and 0.0107 +/- 0.0041 Gy(-1) in the previous study. In the present experiment, irradiations with 144 keV neutrons and 60Co gamma rays were both performed at 21 degrees C, while in the earlier experiment irradiations with 565 keV neutrons were performed at 21 degrees C and the corresponding reference irradiation with gamma rays was performed at 37 degrees C. However, the temperature difference between 21 degrees C and 37 degrees C has a minor influence on the yield of chromosomal alterations and hence RBE values. The large cubic PMMA phantom that was used for the gamma irradiations in the present study results in a larger dose contribution from Compton-scattered photons compared to the mini-phantom used in the earlier experiments. The contribution of these scattered photons may explain the large value of alpha(dic) for gamma irradiation in the present study. These results indicate that the yield coefficient alpha(dic) for 144 keV neutrons is similar to the one for 565 keV neutrons, and that modification of the alpha(dic) value of the low-LET reference radiation, due to changes in the experimental conditions, can influence the RBE(m). Consequently, alpha(dic) values cannot be shared between cytogenetic laboratories for the purpose of assessment of RBM(m) without verification of the comparability of the experimental conditions. PMID- 11893249 TI - Suitability of FISH painting techniques for the detection of partial-body irradiations for biological dosimetry. AB - Peripheral blood was irradiated with 2, 3, 4 or 5 Gy of X rays and was mixed with nonirradiated blood at five different dilutions to simulate partial-body irradiations. Analysis by FISH was performed using whole-chromosome painting probes for chromosomes 1, 4 and 11 in combination with a pancentromeric probe. Chromosome aberrations affecting the painted fraction were classified according to the PAINT nomenclature; other unstable aberrations affecting the unpainted material were also recorded. To evaluate the suitability of painting for dose assessment in partial-body irradiations, the ability of the u test and a proposed s test to detect the expected overdispersion and the similarity between the real doses and the doses estimated using Dolphin's approach were considered. For short term biodosimetry, compared with solid-stained dicentric analyses, the suitability of FISH painting techniques for the detection of partial-body exposures is reduced, because of the decrease in the frequency of aberrations detected by FISH and in the number of cells with two or more aberrations. For reconstruction of past doses, when only complete apparently simple translocations in cells free of unstable aberrations were considered, the detection of the overdispersion and the accuracy of dose estimations were dramatically reduced. In a partial-body exposure, as the original dose increased, the whole-body dose estimated a long time after irradiation would tend to be lower, and the difference from the original dose would tend to be greater. PMID- 11893250 TI - Choosing metaphases for biological dosimetry by fluorescence in situ hybridization (FISH). AB - Data are presented for a subset of lymphocytes characterized by FISH as missing painted chromosomal material. These lymphocytes occur in both control and irradiated subjects. These cells have a much greater frequency of one-way translocations than cells in which all of the painted chromosomal material is present. Their presence contributes to interindividual variability in control translocation yields. These cells do not appear to be more prevalent in persons exposed to high radiation doses. It is suggested that their exclusion when selecting cells for analysis may improve the sensitivity of FISH as a biological dosimeter at low doses. Mechanisms for the production of these one-way translocations in vivo are also discussed, with a proposal that their variable frequency in individuals may be consistent with exposure to chemical clastogens. PMID- 11893251 TI - The micronucleus and G2-phase assays for human blood lymphocytes as biomarkers of individual sensitivity to ionizing radiation: limitations imposed by intraindividual variability. AB - As part of a program to assess the applicability of the micronucleus (MN) and G2 phase assays as biomarkers of cancer susceptibility, we investigated the inter- and intraindividual variations of these end points. For the MN assay, unstimulated blood cultures from 14 healthy donors were exposed in vitro to 3.5 Gy 60Co gamma rays; for the G2-phase assay, PHA-stimulated cell cultures were irradiated with a dose of 0.4 Gy 60Co gamma rays in the G2 phase of the cell cycle. Two of the 14 volunteers were assayed 9 times over a period of 1 year. The repeat experiments revealed that the intraindividual variability was not significantly different from the interindividual variability for both the G2 phase and MN assays. Since the intraindividual variability determines the reproducibility of the assay, our results highlight the limitations of these end points in detecting reproducible differences in radiation sensitivity between individuals within a normal population. For example, one donor of the population was identified as being radiosensitive (based on the 90th percentile criterion) but turned out to be normal when the assay was repeated twice. We conclude that the determination of individual radiosensitivity with these two cytogenetic assays is unreliable when based on one blood sample. PMID- 11893252 TI - Early induction of CDKN1A (p21) and GADD45 mRNA by a low dose of ionizing radiation is due to their dose-dependent post-transcriptional regulation. AB - Previous studies have shown that induction of some genes by low-dose radiation has a different dependence on the time after irradiation than induction by high doses. To examine the mechanisms underlying this phenomenon, we investigated the changes in the time course of the rates of transcription of genes in cells of the human myeloblastic leukemia cell line ML-1 by a nuclear run-on assay. It is possible that the more rapid induction of the mRNA of the CDKN1A and GADD45 genes after exposure to 50 cGy of X rays than after 20 Gy is due to a lower level of stabilization of the mRNA of these genes after 50 cGy. In addition, our results show that 50 cGy of X rays increases the transcription rates of the CDKN1A and GADD45 genes, with a maximum induction at 0.5 to 1 h after irradiation, much earlier than the maximum accumulation of stabilized TP53 protein. We suggest the involvement of BRCA1 protein in the early induction of transcription of these two genes. PMID- 11893253 TI - Feasibility of assessing the carcinogenicity of neutrons among neutron therapy patients. AB - Nuclear workers, oil well loggers, astronauts, air flight crews, and frequent fliers can be exposed to low doses of neutrons, but the long-term human health consequences of neutron exposure are unknown. While few of these exposed populations are suitable for studying the effects of neutron exposure, patients treated with neutron-beam therapy might be a source of information. To assess the feasibility of conducting a multi-center international study of the late effects of neutron therapy, we surveyed 23 cancer centers that had used neutron beam therapy. For the 17 responding institutions, only 25% of the patients treated with neutrons (2,855 of 11,191) were alive more than 2 years after treatment. In a two-center U.S. pilot study of 484 neutron-treated cancer patients, we assessed the feasibility of obtaining radiotherapy records, cancer incidence and other follow-up data, and of estimating patient organ doses. Patients were treated with 42 MeV neutrons between 1972 and 1989. Applying a clinical equivalence factor of 3.2 for neutrons, total average organ doses outside the treatment beam ranged from 0.14 to 0.29 Gy for thyroid, 0.40 to 2.50 Gy for breast, 0.63 to 2.35 Gy for kidney, and 1.12 to 1.76 Gy for active bone marrow depending upon the primary cancer treatment site. We successfully traced 97% of the patients, but we found that patient survival was poor and that chemotherapy was not confirmable in a quarter of the patients. Based on our findings from the international survey and the feasibility study, we conclude that a large investigation could detect a fivefold or higher leukemia risk, but would be inadequate to evaluate the risk of solid cancers with long latent periods and therefore would likely not be informative with respect to neutron-related cancer risk in humans. PMID- 11893255 TI - Tissue engineering and ENT surgery. AB - Tissue engineering is the development of biological substitutes for the repair and regeneration of damaged tissues. We explain the principles of this emerging field of biotechology. The present and potential applications of tissue engineering technologies in ENT surgery are then reviewed. PMID- 11893254 TI - NF-kappa B DNA-binding activity in embryos responding to a teratogen, cyclophosphamide. AB - BACKGROUND: The Rel/NF-kappaB transcription factors have been shown to regulate apoptosis in different cell types, acting as inducers or blockers in a stimuli- and cell type-dependent fashion. One of the Rel/NF-kappaB subunits, RelA, has been shown to be crucial for normal embryonic development, in which it functions in the embryonic liver as a protector against TNFalpha-induced physiological apoptosis. This study assesses whether NF-kappaB may be involved in the embryo's response to teratogens. Fot this, we evaluated how NF-KappaB DNA binding activity in embryonic organs demonstrating differential sensitivity to a reference teratogen, cyclophosphamide, correlates with dysmorphic events induced by the teratogen at the cellular level (excessive apoptosis) and at the organ level (structural anomalies). RESULTS: The embryonic brain and liver were used as target organs. We observed that the Cyclophosphamide-induced excessive apoptosis in the brain, followed by the formation of severe craniofacial structural anomalies, was accompanied by suppression of NF-kappaB DNA-binding activity as well as by a significant and lasting increase in the activity of caspases 3 and 8. However, in the liver, in which cyclophosphamide induced transient apoptosis was not followed by dysmorphogenesis, no suppression of NF-kappaB DNA-binding activity was registered and the level of active caspases 3 and 8 was significantly lower than in the brain. It has also been observed that both the brain and liver became much more sensitive to the CP-induced teratogenic insult if the embryos were exposed to a combined treatment with the teratogen and sodium salicylate that suppressed NF-kappaB DNA-binding activity in these organs. CONCLUSION: The results of this study demonstrate that suppression of NF-kappaB DNA-binding activity in embryos responding to the teratogenic insult may be associated with their decreased resistance to this insult. They also suggest that teratogens may suppress NF-kappaB DNA-binding activity in the embryonic tissues in an organ type- and dose-dependent fashion. PMID- 11893256 TI - Parapharyngeal space tumours: an 18 year review. AB - Parapharyngeal space tumours account for only 0.5 per cent of all head and neck tumours. Due to their inherent location, they present with varied non-specific signs and symptoms, resulting in a delay in diagnosis and unnecessary procedures, such as a 'tonsillectomy' or 'incision and drainage' of a 'quinsy'. Thirty-one patients, operated on over an 18-year period (1981 to 1998), in the Department of Otolaryngology, Singapore General Hospital, are presented. Their ages ranged from 21 to 86 years, with a mean of 52 years, with equal sex distribution. The commonest aetiology was a deep lobe of parotid tumour (44 per cent), followed by neurilemmomas (18 per cent), there was only one paraganglioma. The transcervical and transparotid approaches were the commonest used. The mean surgical time was three hours, mean hospital stay was 5.3 days and post-operative complications were minimal. The average follow-up time was 5.6 years. Although parapharyngeal space tumours are uncommon, recognizing them would enable the correct sequence of investigations, instead of unnecessary procedures resulting in an increased morbidity for the patient. PMID- 11893258 TI - An observational study of the management of traumatic tympanic membrane perforations. AB - Controversies of how best to treat fresh tympanic membrane perforations have always existed. While some otolaryngologists prefer the paper patch method, others prefer modified myringoplasty. A prospective study is needed to investigate the most effective and least expensive management of this common ear trauma. This study examined prospectively, in three sections, a group of patients with a cellophane patch (n = 6), another group with a gentamicin ointment seal (n = 15) and a control group (n = 9) with a gentamicin plug placed at the distal end of the external auditory cavity. Successful healing of the traumatic tympanic membrane perforations was achieved in 50 per cent of the cellophane seal group, 86.7 per cent of the gentamicin ointment seal group and 77.8 per cent of the control group. This study shows that the management of a fresh tympanic membrane perforation should be limited to cleaning the traumatized ear and preventing infection. PMID- 11893257 TI - Vinegar treatment in the management of granular myringitis. AB - To compare the therapeutic efficacy in the management of granular myringitis, 15 patients with chronic granular myringitis were treated with antibiotic ear drops that were used twice to four times a day, and another 15 patients were treated with daily irrigation of the external canal with dilute vinegar solution. All patients treated with dilute vinegar solution had resolution of their original otorrhoea within three weeks, whereas two-thirds of patients recovered within three weeks when treated with antibiotic ear drops. The disadvantages of dilute vinegar therapy were canal irritation with pain and dizziness. When the therapeutic efficacy was compared statistically, a dry ear was attained in the dilute vinegar-treated group at six weeks and six months in the antibiotic ear drop treated group (p<0.01). These results suggest that very low pH therapy using dilute vinegar solution is definitely effective in the management of granular myringitis. PMID- 11893259 TI - Functional endoscopic sinus surgery in the treatment of massive polyposis in asthmatic patients. AB - The association between asthma and sinonasal disease has been known for years. Effective treatment of sinonasal disease, which is one of the factors that exacerbate asthma, may also improve and stabilize the asthmatic condition. This study examines the outcome of functional endoscopic sinus surgery (FESS) on asthmatic patients with massive nasal polyposis. Thirty-four asthmatic patients were included in the study. All were operated on in our department and were analysed for pre-operative data regarding their asthma and sinonasal disease. A questionnaire regarding subjective evaluation of asthma and sinonasal status was presented to the patients, and objective evaluations, including nasal endoscopy and spirometry, were performed. Follow-up endoscopy revealed satisfactory results in 88 per cent, with positive correlation to the patients' subjective assessment of nasal status. No such correlation was found with regard to subjective and objective assessment of asthma: a small group of patients had completely clean sinonasal cavities with no perceived improvement in their asthmatic condition. The use of prednisolone and bronchodilators was significantly reduced post operatively. However, in a subgroup of 13 patients followed at the asthma clinic, who had adequate pre-operative and post-operative data, there was no difference in their pre- and post-operative asthma condition. Seven had minimal improvement and in six there was a definite worsening of their asthma; nevertheless, nasal breathing and quality of life improved in most patients. The mean follow-up was 2.1 years. Thus, we conclude that in this study FESS does not improve asthma, but does improve the quality of the life of the patient. PMID- 11893262 TI - Applicability of the adult comorbidity evaluation - 27 and the Charlson indexes to assess comorbidity by notes extraction in a cohort of United Kingdom patients with head and neck cancer: a retrospective study. AB - The term comorbidity stands for disease processes that co-exist and are not related to the index disease being studied. Comorbidity in cancer has been shown to be a major determinant in treatment selection and survival. Patients with head and neck cancer can have significant comorbidity owing to the high incidence of tobacco and alcohol abuse. No studies to date have addressed this problem in head and neck cancer patients in the United Kingdom. The applicability of the adult comorbidity evaluation - 27 index (ACE-27) and the Charlson index (CI) to assess the comorbidity burden by retrospective notes review is studied here. Retrospective data collection and completion of a comorbidity index in a United Kingdom setting is feasible. We conclude that the pre-assessment visit is a useful time to record comorbidity and as a significant amount of information required for grading relates to historical items, this is best done using a self administered patient questionnaire. PMID- 11893261 TI - Computed tomography and ultrasonographic evaluation of metastatic cervical lymph nodes with surgicoclinicopathologic correlation. AB - The detection of cervical lymph nodal metastasis and carotid artery invasion by metastatic lymph nodes is an important issue in the management of head and neck malignancies. This study compared the evaluation of metastasis by palpation, ultrasonography (USG) and computed tomography (CT) in patients with known head and neck malignancies. Twenty-five consecutive patients with head and neck malignancy were prospectively evaluated for the presence of cervical lymphadenopathy and carotid artery invasion. All patients underwent clinical examination (palpation), USG and CT examination. A modified CT criteria was employed which yielded acceptable results for the detection of metastatic nodes. Radical neck dissection was performed for 26 neck sides, and the results of pre operative evaluation were confirmed by the surgical and histopathological findings. Palpation, ultrasound and CT have comparable sensitivity in the determination of metastasis involving cervical lymph nodes. Thus palpation should be employed as the primary method of assessment of secondaries in the neck. However, palpation is less sensitive than CT and USG in the detection of carotid artery involvement, hence the clinical suspicion of arterial invasion should be confirmed by either CT or USG which have similar accuracy in the detection of carotid artery invasion. PMID- 11893260 TI - Second malignancies in early stage laryngeal carcinoma patients treated with radiotherapy. AB - A retrospective review of 240 patients with T1/T2 squamous cell carcinomas of the larynx was performed. Seventy-two per cent had glottic primaries, 27 per cent had supraglottic tumours and one per cent had subglottic disease. Sixty-nine per cent presented with T1 disease and 31 per cent had T2 staged tumours. All patients were treated with definitive radiotherapy between 1973 and 1997. With a median follow-up of 68 months, 68 patients (28 per cent) have developed 72 other cancers. Ten of 68 presented with synchronous primaries (15 per cent). Thirty per cent of glottic patients and 25 per cent of the supraglottic/subglottic patients developed second cancers. The most frequent second malignancy was lung cancer: 28/72 (39 per cent). Fifteen patients developed second head and neck cancers (21 per cent). Other second primary sites included oesophagus (eight), prostate (six), colorectal (five), breast (two) and others (eight). The median time from radiotherapy until the development of a second cancer was 31 months. The Kaplan Meier survival estimate at five years was significantly less for those patients developing second cancers (55 per cent) compared to those not developing second malignancies (70 per cent), (p<0.05). The median survival from the development of a second cancer was 14 months. More died as a result of a second cancer (41 patients) than their primary laryngeal cancer (40 patients). Second cancers are common and deadly in patients with early stage laryngeal carcinoma. PMID- 11893263 TI - How I do it; securing a full thickness skin graft. AB - The authors present a novel way of securing a full thickness skin graft (Wolfe graft) that we believe is not only time saving but also helps to create a dressing that exerts an equal pressure over all areas of the graft. PMID- 11893264 TI - Spherical foreign bodies in the oesophagus removed by balloon angiographic catheter. AB - Two children aged three and seven years presented to the department of Otolaryngology with total dysphagia following the accidental swallowing of a steel ball bearing and a plastic ball. These rare spherical foreign bodies were removed successfully by oesophagoscopy under general anaesthesia using an innovative method involving a balloon angiographic catheter. PMID- 11893265 TI - Computerized tomography is not reliable in the diagnosis of brainstem infection. AB - The case of a 17-year-old girl who presented with a two-day history of absolute dysphagia secondary to a bulbar palsy due to a pre-pontine abscess is described. Rigid oesophagoscopy was normal and a neurology consultation suggested a central cause for her dysphagia. However the diagnosis was delayed because a computed tomography (CT) scan of her brain and brainstem was reported as normal. A subsequent magnetic resonance image (MRI) scan revealed a pre-pontine abscess. CT scanning is not as reliable as MRI in the diagnosis of infective lesions of the brainstem/brain, especially early in the course of the infection. PMID- 11893266 TI - Long-term follow-up after laser-induced endotracheal fire. AB - The objective of this presentation is to outline long-term complications and their management in contrast to acute measures after endotracheal laser-induced fire. This case focuses on a 56-year-old patient in whom an endotracheal fire occurred during CO2 laser surgery. Despite local swelling and evidence of acute lung injury, the patient was extubated the following day under single-shot cortisone and inhalation of dispersed adrenaline under assisted spontaneous breathing. Wound healing was assessed by regular flexible bronchoscopy and spirometry. Fourteen weeks after uneventful recovery, the patient presented with acute inspiratory stridor, related to a tracheal stenosis 2.5 cm distal to the glottic level. After tracheal end-to-end anastomosis, further follow-up was uneventful. Early extubation under ITU conditions avoided the need for tracheostomy and its sequelae. However, tracheal stenosis did not become apparent before week 14. While in acute management of laser-induced endotracheal fire a conservative approach was established successfully, the risk of further long-term complications implies the need for a prolonged follow-up regime even in cases of less extensive burns. PMID- 11893267 TI - Lemierre's syndrome as a consequence of acute supraglottitis. AB - Lemierre's syndrome comprises internal jugular vein thrombosis following oropharyngeal sepsis and is a rare and serious condition. It is most commonly caused by the anaerobe Fusobacterium necrophorum and typically presents as metastatic sepsis to the lungs and joints. Thrombosis is demonstrated by computed tomography (CT) of the neck, and it is routinely treated with intravenous antibiotics and anti-coagulation. We describe a case of Lemierre's syndrome following acute supraglottitis. The clinical features were of retrograde intracranial thrombosis, rather than the more usual metastatic sepsis. PMID- 11893268 TI - First case of full-thickness palatal necrotizing sialometaplasia. AB - Necrotizing sialometaplasia is an uncommon, benign, self-limiting condition which can stimulate malignancy. The commonest site of occurrence is the hard palate. We report the first case with full thickness palatal involvement. The clino pathological features of this condition are discussed. PMID- 11893269 TI - Cowden's disease: a rare cause of oral papillomatosis. AB - Cowden's disease is a rare autosomal dominant condition with characteristic mucocutaneous papillomatous lesions. These lesions are mucocutaneous markers for increased risk of malignancies in the thyroid, breast and the gastrointestinal tract. We discuss the case of a 50-year-old female patient who presented with oral and cutaneous papillomoas and a past history of breast malignancy. Important management aspects of these patients are considered. PMID- 11893270 TI - Synovial sarcoma in the retropharyngeal space. AB - Synovial sarcoma is an aggressive mesenchymal tumour, rarely occurring in the head and neck. Management guidelines are by extrapolation of management of sarcomas in the extremities. We present a case involving the retropharyngeal space in a 20-year-old male. Analysis of more data on head and neck synovial sarcoma is necessary to make meaningful management recommendations. PMID- 11893271 TI - Bilateral cortical blindness: an unusual complication following unilateral neck dissection. AB - We present the case of a 50-year-old man who developed bilateral cortical blindness and confusion following a seemingly uneventful right-sided radical neck dissection. Computed tomography (CT) scans confirmed bilateral occipital lobe infarctions. To our knowledge, there are no previously documented reports of this clinical event following head and neck surgical procedures. Although this is a rare occurrence, otolaryngologists should be aware of this potential post operative complication. The possible aetiologies of this condition are discussed. PMID- 11893272 TI - 'Horseshoe-shaped' post-operative alopecia following lengthy head and neck surgery. AB - We report a case of 'horseshoe-shaped' pressure-induced post-operative alopecia following a lengthy head and neck procedure. Post-operative hair loss is rare and to our knowledge has only previously been found in fields of surgery where careful head positioning is not an inherent part of the procedure. In these cases there has been a single area of hair loss from the central occipital area and per operative pressure effects of the head resting on the operating table have been postulated as the likely cause. The case presented shows an area of hair loss closely corresponding to the shape of the head rest used during a long procedure. This strongly supports the theory that prolonged pressure is the likely cause. The mechanism of pathogenesis is discussed together with a suggested strategy for its avoidance. The single most important aspect of prevention of this complication of surgery is the knowledge of its existence and aetiology. PMID- 11893273 TI - Auto-immune thyroiditis presenting as a thyroglossal tract swelling. AB - Both thyroglossal cysts and aberrant thyroid tissue may present as a mid-line neck swelling. We report a case of autoimmune thyroiditis presenting as a thyroglossal swelling in a middle-aged woman. This very rare finding is discussed with reference to the optimal management of thyroglossal tract swellings. PMID- 11893274 TI - The effect of treatment of twin-twin transfusion syndrome on the diagnosis-to delivery interval. AB - No randomised controlled trials of treatment of twin-to-twin transfusion syndrome (TTTS) exist. Since severely preterm neonatal survival has increased over time, survival as an outcome measure is confounded by improvements in neonatal care. The diagnosis-to-delivery interval is a measure of success of treatment independent of improvements in neonatal care. We wished to evaluate whether treatment of TTTS is associated with a lengthening of the diagnosis-to-delivery interval. MEDLINE search was performed supplemented by careful reference review. All TTTS series were included where the following information on each patient was available: survival, fetal demise, gestational age at diagnosis and diagnosis-to delivery interval in days. INCLUSION CRITERIA: gestational age at diagnosis < 29 weeks and diagnosis by ultrasound in the absence of maternal symptoms. Cases undergoing multiple types of treatment were excluded. Eight publications met inclusion criteria and included the following cases: controls (n = 16), amnioreduction (n = 61), septostomy (n = 12), and fetoscopic laser occlusion of chorioangiopagus vessels (n = 51). There was no difference in the diagnosis-to delivery interval, overall survival, at least one survivor, or number of fetal deaths between the four groups. Logistic regression using at least one survivor as the dependent variable revealed a positive association with gestational age at diagnosis and with diagnosis-to-delivery interval, a negative correlation with fetal death, and no correlation with treatment group. We conclude that there is no difference in diagnosis-to-delivery interval or survival for any treatment for TTTS compared to expectant management. The lack of significance appears to be due to small sample sizes. PMID- 11893275 TI - Rejoinder to meta-analysis of twin-twin transfusion by Skupski et al. AB - Assessment of treatment options in twin-twin transfusion should involve a detailed interrogation of the placenta and fetuses prior to treatment, stratification according to response to amnioreduction and careful analysis of quality of survival. Amnioreduction and laser coagulation are not alternative and equivalent modalities for the treatment of twin-twin transfusion. Randomized trials may not be ethical without prior therapeutic/diagnostic amnioreduction. PMID- 11893276 TI - Hypertensive disease in twin pregnancies: a review. AB - Reports over the past seventy years show that twin gestations lead to an increased risk of hypertensive disorders. Numerous studies discuss the incidence of hypertensive disease in twin versus singleton gestations, as well as effects of parity, race, age, income level, smoking, zygosity and heritability on this condition. The range of relative risk of gestational hypertension, preeclampsia and eclampsia for twin compared to singleton gestations is 1.2 to 2.7, 2.8 to 4.4 and 3.4 to 5.1 respectively. Parity, African-American ethnicity, and young maternal age are all factors that increase the relative risk of acquiring hypertensive disease to 4.0, 1.8 and 1.5 in mothers of twin gestations. Factors such as maternal smoking, income level and zygosity have a negligible effect on the relative risk of acquiring hypertensive disease in twin gestations. In addition to twin mothers exhibiting a higher incidence of hypertensive disease compared to their singleton counterparts, they also exhibit an earlier onset of hypertensive disease at both 35 and 37 weeks of gestation comparatively. Uric acid levels measured at 30-31 weeks of gestation in twin mothers predicted the onset of preeclampsia with a sensitivity of 73% and a specificity of 74%. The range of risks presented in the literature is wide and the therapies avocated are diverse. We therefore decided to summarize the risks in a comparative fashion and to review current therapeutic strategies for the convenience of clinicians who confront increasing numbers of multiple pregnancies. The tables bring all recent published risks together in the first comparative analysis in which the data has been converted to relative risks and confidence intervals. Because the literature is relatively silent on specific management of hypertensive disease in twin pregnancies, general management recommendations for singleton gestations should be used by practitioners caring over twin gestations. PMID- 11893277 TI - Twinning and birth weight in the Israeli Jewish versus Muslim maternities. AB - Ethnicity differences account for genetic, environmental, lifestyle, and reproductive variables, influencing the rate of twinning (Nylander, 1981). Frequently, ethnic differences correlate with variable perinatal care leading to differences in outcome. Free access to antenatal care, and to facilities for delivery and neonatal care is available for the entire population in Israel, and therefore differences attributed to levels of medical care are practically negligible. We previously evaluated the overall relationship between ethnicity and outcome in a population-based cohort of mothers of twins (Goldman et al., 2001). However, the overall comparison may have masked some differences that could be present. The purpose of this study was to evaluate whether ethnicity is associated with differences in perinatal outcome in randomly selected, matched controlled Israeli Jewish and Muslim mothers of twins. PMID- 11893278 TI - On the standardisation of the twinning rate. AB - In many studies the twinning rate, being strongly dependent on maternal age (and parity), has been standardised according to the maternal age distribution. The direct method requires very informative twinning data for the target population. The indirect method is used when the data for the target population is not sufficiently informative or when the target population is small. We have earlier introduced an alternative indirect technique for standardising the twinning rate. Our technique requires even less of the twinning data. Besides maternal age, parity is an influential factor, and should, if possible, be taken into account. In this study we present the traditional standardisation methods based on both maternal age and parity, we propose a new direct standardisation method and we develop our standardisation methods so that they take into account both maternal age and parity. We apply these standardisation methods to data from Finland, 1953 1964, from St. Petersburg, Russia, 1882-92, from Canada 1952-1967, and from Denmark, 1896-1967. These methods all give results very similar to those for the Finnish data, but the effect of parity is strongest with the direct methods. This may be due to the fact that, among extramarital maternities, parity has a strongly increasing effect on the twinning rate. This may be attributed to a higher reproduction capacity among unmarried mothers. Standardisations of the Canadian and the Danish data also give reliable results. With the St. Petersburg data, however, the different standardisations show notable discrepancies. These discrepancies are compared with Allen's findings. PMID- 11893279 TI - Gene-environment interaction effects on behavioral variation and risk of complex disorders: the example of alcoholism and other psychiatric disorders. AB - There have been few replicated examples of genotype x environment interaction effects on behavioral variation or risk of psychiatric disorder. We review some of the factors that have made detection of genotype x environment interaction effects difficult, and show how genotype x shared environment interaction (GxSE) effects are commonly confounded with genetic parameters in data from twin pairs reared together. Historic data on twin pairs reared apart can in principle be used to estimate such GxSE effects, but have rarely been used for this purpose. We illustrate this using previously published data from the Swedish Adoption Twin Study of Aging (SATSA), which suggest that GxSE effects could account for as much as 25% of the total variance in risk of becoming a regular smoker. Since few separated twin pairs will be available for study in the future, we also consider methods for modifying variance components linkage analysis to allow for environmental interactions with linked loci. PMID- 11893280 TI - The assessment of peer selection and peer environmental influences on behavior using pairs of siblings or twins. AB - Many studies have found strong peer correlations for a variety of problem behaviors that begin in adolescence (e.g. substance use). Such correlations are commonly attributed to peer influences, but could also be explained by selective ('assortative') friendship: the tendency for those with similar patterns of behavior to become friends. Here we show how, under certain assumptions, cross sectional data from pairs of siblings or twins and their peers may be used to resolve the contributions of peer selection and reciprocal peer environmental influences to peer resemblance. We performed power calculations to determine necessary sample sizes for rejecting with 80% power, at the 5% significance level, the hypothesis of only peer selection effects, or only reciprocal peer environmental effects. A false hypothesis of only selective friendship effects was always easier to reject than a false hypothesis of only reciprocal peer environmental influences. Limitations of these simulations, including uncertainty about the most appropriate way to model peer selection, are discussed. PMID- 11893281 TI - Why do identical twins differ in personality: shared environment reconsidered. AB - While heritability studies show that most of the variance in adult personality can be attributed to genetic or so-called nonshared environmental influence, this does not mean that shared events lack importance for the development of later personality differences. We studied the relationship between Big Five personality differences in monozygotic (MZ) twins at age 29, and life stressors at age 6 to 15, using prospective data from 26 MZ pairs studied from birth onwards. A positive significant correlation was found between stressors in childhood and early adolescence, and intrapair personality differences in Agreeableness, Openness, Conscientiousness, and five-factor profiles. We note that the effects of shared events are labeled "nonshared" environment when the effect is to make siblings more different. Case examples illustrate the relationship between stress and personality differences, and provide hypotheses for further studies in larger samples. PMID- 11893282 TI - Russian twin studies: colleagues, controversies, case studies and current events. AB - The 2001 honoring of Russian twin researcher, Dr. Inna V. Ravich-Shcherbo, at the Tenth International Congress on Twin Studies (ISTS), in London, brought timely recognition to an international colleague. It also marked an occasion for reflecting on the course of twin studies in Russia. Historical trends and current accomplishments are examined with an eye toward future developments. Next, the distinguished careers of Russian monozygotic twins, Drs. Alexander and Andrew Fingelkurts, exemplify twin research findings on intelligence and occupational choice, and illuminate the status of twin studies and other scientific research in their country, are told. Their life histories are followed by the initially tragic, but ultimately heartwarming, story of young twins, Max and Andy, whose physical disabilities were overcome through the efforts of an empathic war veteran. The recent Moscow Summer School, the first in a three-time lecture series, encouraged crucial academic exchange among scientific investigators and students from around the world. Final thoughts are that much can be learned and much can be accomplished, given that we continue to nurture the twin-related resources available to us. PMID- 11893284 TI - [P300 clinical utility in major depression]. AB - INTRODUCTION: Event related potentials are an objective parameter reflecting cognitive functions. Among the event related potentials, the P300 component is viewed as a measure of stimulus evaluation time and it can provide a rough estimate of the time required for perceptual processing. Impairment in cognitive processing, psychomotor retardation and abnormally amplitude and longer P300 latency have been found in depressive patients. METHODS: To evaluate the influence of visual and auditory stimuli on the P300 latency we studied 595 patients with major depression. The experimental tasks applied were, first, a series of 300 auditory stimuli: 85% were tones of 1,000 Hz, and 15% were tones of 2,000 Hz, and second, a series of 300 visual stimuli; 85% were black circles on a white background and 15% were black squares on a white background. RESULTS: The results shown an increase of P300 latency in depressive patients during auditory and visual tasks. DISCUSSION: These results are consistent with an impairment in brain function with cortical hypo activity in depressive patients that is associated with cognitive deficit processing. These results determine the clinical utility of P300 in patients with major depression. PMID- 11893286 TI - [Incidence of anorexia nervosa in a community mental health services for children and adolescents]. AB - INTRODUCTION: In the last few years, a huge social importance is being given to eating disorders and specially to anorexia nervosa. Sometimes, the actual epidemiology doesn't correspond with that voice of alarm. MATERIAL AND METHODOLOGY: In this article are reviewed a sample of 3,849 clinical histories (aged 0 to 16 years) collected in 9 years in a public children and adolescents mental health unit.Objective. To see the incidence of the eating disorders (with their characteristics of sex and age) and the progression of cases of anorexia. RESULTS: Show that the percentage of eating disorders (including anorexia) compared with the rest of psychopathologies is very low, that the cases of eating disorders have risen in the last 4 years and that the cases of anorexia nervosa haven't risen in the last decade. DISCUSSION: Given the discrepancy with other authors who found several cases, we look for an interpretation of our data. PMID- 11893287 TI - [Cognitive constructs related to depression in children]. AB - Hopelessness, anhedonia and automatic thoughts have all been related to major depression and depressive symptoms. Few research has been done on these constructs in children, specially together. AIM: The aim of this study was to translate into Spanish and validate the hopelessness scale for children, the pleasure scale for children, the automatic thoughts scale and the Kovak's child depression inventory, and then to relate the cognitive constructs to depressive symptoms in a sample of normal children. METHODS: Children were recruited from the Federal Primary School Mexican Air Forces of Mexico City. All Children from the 3rd to 6th grades were included. RESULTS: 256 children were included, 53% male and 46% female, mean age 9.8 1.15 years. Internal consistencies for instruments were as follows: anhedonia scale 0.91, hopelessness scale 0.63, automatic thoughts 0.87 and depression 0.80. The factor structures of the translated versions were similar to those reported by their original authors. Interinstruments correlations ranged from 0.17 to 0.39. Automatic thoughts and depressive symptoms reached the highest significant value (r= 0.39). 6th graders rated significantly lower in all instruments than their 5th and 3rd peers. CONCLUSIONS: All measures studied demonstrate moderate to high internal reliability and the factor structures predicted. Depressive symptoms among normal children related weakly but significantly to the cognitive constructs studied, just as seen in normal adults. PMID- 11893285 TI - [Spanish version of the personality diagnostic questionnaire-4+ (PDQ-4+)]. AB - INTRODUCTION: The Personality Diagnostic Questionnaire is one of the most frequently used instruments for diagnosing personality disorders (PD). Its last version (PDQ-4+) combines the easy applicability of a questionnaire with the control of state symptoms interference of an interview. The aim of this paper is to present preliminary results on the Spanish version of the PDQ-4+. METHOD: 159 psychiatric outpatients were assessed using the questionnaire of PDQ-4+. Among them, a sample of 47 were also evaluated using the short interview of clinical significance that completed the PDQ-4+. RESULTS: Results obtained were very similar to that from previous research, indicating similar psychometric properties of the PDQ-4+ Spanish version, to English, Italian, Chinese and Norway versions. In general, the internal consistency was acceptable; the obsessive compulsive PD scale showing the lowest Cronbach's alpha. The high prevalence of PD obtained with the PDQ-4+ questionnaire was dramatically reduced when the clinical significance scale was added. These results are in more agreement with previous epidemiological studies. DISCUSSION: Overall, the PDQ-4+ questionnaire seems a good screening instrument in clinical use. Best results are obtained when the clinical significance scale is also administered. PMID- 11893288 TI - [The Spanish version of the Manchester Nurse Rating Scales]. AB - INTRODUCTION: There is no Nursing-staff Mania Rating Scale adapted to the Spanish language. The present work is the result of the adaptation we have carried out. METHODOLOGY: The scale has been translated and the back-translation compared with the original version. It has been applied on 670 occasions to a sample of 59 patients. Simultaneously the patients were rated with the Manic Interpersonal Interaction Scale, two versions of the Visual Analogue Scale and the Numeric Rating Scale. RESULTS: The coefficient Alpha of Cronbach is 0.94. The interrater's correlation coefficient for the total of the scale is 0.84. The factorial analysis gives a single factor that explains 65% of the variance. The Area under the ROC Curve is 0,87. CONCLUSIONS: The scale shows excellent psychometric properties, with great homogeneity of the scale items and good discriminate properties between bipolar patient and those affected by other psychoses. PMID- 11893289 TI - [Validation of a version in Spanish of the Yale-Brown Obsessive-Compulsive Scale]. AB - INTRODUCTION: The objective of this study is to validate a version in Spanish of the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS). METHOD: We administered to 20 patients with obsessive-compulsive disorder (OCD) the Maudsley Obsessive Compulsive Inventory (MOI) and a version in Spanish of the Y-BOCS. The application of the last one was performed by an interviewer, filmed, and then watched and rated by other 4 observers. In addition, the opinion of the treating psychiatrist about the severity of the patient's symptoms was registered. The interviewer and the observers were named raters. RESULTS: The mean scores of the Y-BOCS assigned by the 4 observers were from 18.35 to 19.55; the maximum scores, from 34 to 35; and the minimum scores, from 1 to 5. The Pearson's correlations between raters were significant for all cases (r>0.86 and p<0.001). For all raters, Y-BOCS scores significantly correlated with MOI scores (p<0.001) and the opinion of the treating psychiatrist (p<0.05). We did not find significant correlation between the last one and MOI scores. The Cronbach's coefficients ranged from 0.8789 to 0.9488. CONCLUSIONS: The Spanish version of the Y-BOCS is reliable, homogeneous and valid for quantification of the obsessive-compulsive symptoms, and has more correlation with the opinion of the treating psychiatrist than the MOI. PMID- 11893290 TI - [The father image in male substance users]. AB - INTRODUCTION: Due to the fact that various research studies have shown that drug users usually have a father with negative characteristics, it is convenient to investigate this phenomenon and take it into consideration in addict treatment efforts. METHODOLOGY: This is a qualitative study that explores the fatherly perception among male substance users. The sample included 25 users who asked for treatment at Youth Integration Center and 25 non-users. Both groups were subjected to projective tests. RESULTS: The father image among substance users emphasizes negative characteristics; in contrast, non-users present a basically positive father perception, considering him as the most valuable family member. CONCLUSIONS: These data show that substance users relate less with the fatherly figure than non-users. They perceive their father as a devaluated member that occupies a secondary place in the family and sometimes does not even have an affective role. The findings of this investigation can be applied in individual psychotherapy, facilitating the revaluation and reflection of the fatherly figure among substance users. It would be convenient to create the necessary conditions to restitute the father role in family therapy and counseling groups for relatives. The findings must consider the size of the sample and limitations of qualitative research methology. PMID- 11893291 TI - [Depressive pseudodementia: Diagnostic frontiers]. AB - In the limits between dementia and depression we find a clinical entity in permanent controversy; Depressive Pseudodementia. Depressive pseudodementia is defended by some authors and criticised by others and set us in the diagnostic frontier palced between the cognitive and the affective, between neurology and psychiatry. The entity is not well delimited and reaches diagnostic relevance clinical, evolution and response aspects. In the present issue, we realize a reflection on the term depressive pseudodementia with a review of the scientific literature with special attention in clinical and diagnostic subjets. We conclude that depressive pseudodementia although considering diagnostic limitations is still a valid term in clinical practice, and it eases the approximation, diagnosis and treatment of patients with mixed symptoms of cognitive and depressive type. PMID- 11893293 TI - [First Ibero-American Sumit on Family Medicine: pooliong forces, sharing experiences]. PMID- 11893292 TI - [Cortico-orbital frontal functions and psychophysiological features of DSM-IV personality disorders]. AB - The purpose of the study was to analyze the construct validity of DSM-IV categories and clusters for personality disorders, using computerized neuropsychological measures exploring frontal executive functions, such as vigilance or sustained attention, mental flexibility, planning and concept formation (Stroop, CPT, WCST); and psychophysiological records of heart rate and skin electric response to experimental stress, and recovery slope in both measures. The sample consisted of 138 participants (66 males and 72 females), with ages between 17 and 65 years, which received a diagnosis of any personality disorder, according to DSM-IV criteria. The results exclusively confirm, to some extent, the construct validity of cluster A, mainly based on neuropsychological deficits, but differences between categories were much more diffuse. PMID- 11893294 TI - [Cardiovascular risk and glucose metabolism: agreements and discrepancies between the WHO-85 and ADA-97 classifications]. AB - AIM. To identify the differences between coronary heart disease risk in patients with altered basal glucemia (ABG), oral glucose intolerance (OGI) and type II diabetes mellitus according to the WHO-85 and ADA-97 diagnostic classifications, in an adult population at high risk for diabetes mellitus. DESIGN: Descriptive, cross-sectional, multicenter study.Setting. Seven primary health care centers in Spain.Patient. 970 persons considered the population at risk for type II diabetes mellitus. MEASURES: Participants were classified according to the criteria of the WHO-85 (normal, OGI, diabetes) and the ADA-97 system (normal, ABG, diabetes). The following variables were recorded: age, sex, smoking habit, body mass index, systolic blood pressure, diastolic blood pressure, basal glucemia, glucemia 2 h after an oral glucose tolerance test, HbA1c, microalbmuniuria, total cholesterol, HDL, LDL and triglycerides. Coronary heart disease risk was calculated with the 1998 table developed by Wilson et al. on the basis of the Framingham study. RESULTS: A total of 970 participants were studied. Mean age was 58.6 #+ 12.4 years; 453 were men (46.7%) and 517 were women (53.3%). Our analysis showed that cardiovascular disease risk factors were less frequent in normal subjects, and that their prevalence was higher in persons with diabetes (according to both WHO and ADA classifications). There were no significant differences in coronary heart disease risk or different risk factors between analogous groups in the two classification systems (normal, OGI/ABG or diabetes). Coronary heart disease risk in persons with different types of alterations in glucose metabolism was 11.3% in normal subjects, 14% in persons with OGI and 27.3% in persons with diabetes according to the WHO-85 system, and 11.4% in normal subjects, 15.7% in persons with ABG and 29.5% in persons with diabetes according to the ADA-97 system. CONCLUSIONS: The greater the alteration in carbohydrate metabolism, the greater the coexistence of risk factors and the estimated risk of coronary heart disease. There were no significant differences in the presence of cardiovascular risk factors, or in the relationship between carbohydrate metabolism and coronary heart disease risk, between analogous stages identified with one classification system or the other. PMID- 11893296 TI - [Life-styles determining the oral health of adolescents in Vitoria (Gasteiz):and evaluation]. AB - OBJECTIVE: To assess the factors determining oral health in adolescents in school at Vitoria (Gasteiz).Design. Descriptive epidemiological study. PARTICIPANTS: Randomised representative sample of 1,380 students from public and private schools. Setting. Vitoria (Gasteiz). MAIN MASUREMENTS AND RESULTS: A questionnaire on bucco-dental health was self-administered. 41.9% of the adolescents brushed their teeth three times or more a day (2.2 ( 0.98 times a day), with greater frequency in girls. 64% never used dental floss, and 30% did so only at times. 88.1% did not complement their oral hygiene with mouth-washes. Consumption of commercial cakes and pastries was 0.74 ( 1.12 times a day; and of sweets, 1.58 ( 3.15 times a day. 81.9% of the adolescents valued their mouth and dental health the same as the health of other organs of their bodies. 63.5% did not know whether any compound was added to the water as a preventive measure against dental disease; and 83.2% of those who thought something was added did not know what compound it was. 66.7% had been to the dentist during the previous year; 28.2% had not been for over a year. CONCLUSION: The oral health habits of adolescents in Vitoria (Gasteiz) show deficiencies that could be corrected through Health Education Programmes and promotion of the use of preventive dental services. PMID- 11893297 TI - [Tackling tobacco dependency among a Health District's employees]. AB - OBJECTIVE: To find the view of health and ancillary staff about tobacco consumption in Health Centres, smokers' profiles, and their degree of motivation to give up. DESIGN: Observational and crossover study.Setting. Primary Care district in the city of Barcelona (Eixample District). PARTICIPANTS: All the health and ancillary staff in the District (n=932). MEASUREMENTS: Personal questionnaire with anonymous reply on: personal details, tobacco habit, view on the function of example, compliance with regulations in force, and their attitude to tobacco in the work-place. MAIN RESULTS: 52% replied; 68.4% were women, with average age 46.9 (SD 9.9). 26.5% smoked; 43.8% had never smoked and 29.6% were ex smokers. 87.5% thought example an important function; 68.1%, that prohibition signs were insufficient; 72.8%, that legal regulations were not complied with; 77.2% thought space should be reserved for smokers; and 61.2% were annoyed that there was smoking in the centre. 63.9% of the health staff counselled smoker patients as a matter of course. There was greater prevalence of smokers among clerical staff, nurses and the "others" group. Smokers consumed 16.6 cigarettes per day (SD 9.5), had been smoking for 23.2 years (SD 9.6). 54.3% had tried at least once to give it up. 59.1% had high dependency and 76.7% had high motivation. 64.8% would participate in activities to stop smoking and 72.4% thought that their employers should provide assistance. CONCLUSIONS: Those surveyed thought that example was an important function, signs were insufficient and the regulations in force were not respected. Half the smokers would be interested in taking part in activities to help them give up and think the company should help them. PMID- 11893298 TI - [Cell anomalies in the cervix and subsequent pre-cancerous lesions in a Health Area]. AB - OBJECTIVE: To determine the frequency of epithelial anomalies in cervical smears during the three years from August 1997 to July 2000 and of subsequent pre cancerous lesions that are histologically confirmed, with differentiation also being made between health care levels. DESIGN: Retrospective descriptive study. SETTING: Anatomy Pathology referral Laboratory. Talavera de la Reina Health Area. PARTICIPANTS: 5,712 cervical smears and 70 biopsies.Intervention. Review of path reports. MAIN MEASUREMENTS AND RESULTS: 308 (5.4%) smears showed anomalies that were not attributed to a benign process. 209 (68%) of these were followed up, 139 (45%) of them through repetition of the smear and 70 (23%) through biopsy. Cell and cytology correlated in 42 women (30.2%), and cell-histology in 27 (38.6%) (0.47%, n = 5,712). 2,874 of the smears in the Area were taken in Primary Care, and 127 (22.2%) cell anomalies and 6 (0.1%) pre-cancerous lesions were found at this health care level. CONCLUSIONS: The 5.4% frequency of anomalies is consistent with that in other population groups with an incidence of cervical cancer similar to ours. The 0.47% pre-cancerous lesions confirmed histologically is very low in comparison with the anomalies, which suggests that this intervention needs to be improved. This could be done by increasing biopsies for cell anomalies and screening of women at risk. Interpretation of smear reports, more information to the women and follow-up of the findings could all be improved, too. PMID- 11893299 TI - [Are COX-2 inhibitors indicated for patients at risk?]. PMID- 11893300 TI - [The family doctor and pregnancy monitoring in the various Autonomous Communities]. PMID- 11893301 TI - [Palliative care. Multidimensional assessment of patients and their families]. PMID- 11893302 TI - [Why do alkaline phosphatases increase?]. PMID- 11893303 TI - [Advanced tools for searching for medical information on the web]. PMID- 11893304 TI - [The home care service: a resource for people with Alzheimer's disease?]. PMID- 11893305 TI - [Ivermectin in scabies treatment]. PMID- 11893306 TI - [The next stage: motivation, resources and cooperation to respond to the population's health needs]. PMID- 11893307 TI - [Clinical managers and line managers in Primary Care]. PMID- 11893309 TI - [Toxic oil syndrome and Paracetamol]. PMID- 11893310 TI - [The epidemic of heart failure]. PMID- 11893311 TI - [Physiology is back in the Cath Lab! Should we abandon angiography in the assessment of intermediate coronary stenosis?]. PMID- 11893312 TI - [Heart failure mortality in Spain, 1977-1998]. AB - INTRODUCTION AND OBJECTIVES: Heart failure is now the third leading cause of cardiovascular death in developed countries and is also an important cause of morbidity and hospitalization that now represents the main cause of admissions among the elderly. In this study we present heart failure mortality trends in Spain developing over the last 20 years. METHODS: Data on deaths due to heart failure were obtained from files supplied by the Spanish National Institute for Statistics. We present age-adjusted specific mortality rates over time analyzed by sex and geographic area. Poisson regression models were used to estimate trends. RESULTS: Heart failure is responsible for 4 to 8% of all-cause mortality in men and women, and for 12 to 20% of cardiovascular mortality overall, the the highest rates seen among the elderly and in Andalusia. The lowest rates are found in the Basque Country and some provinces of Castilla-Leon. Rates have tended to decrease over the last 20 years, but the rate of decrease has been slower in women, such that their mortality began to exceeded that of men from 1990 onwards. Mortality among the elderly has not changed significantly but the total number of deaths and morbidity are both increasing. CONCLUSIONS: Because the Spanish population is aging, we can foresee that chronic heart failure will require greater attention in the future. PMID- 11893313 TI - [External and internal electrical cardioversion: comparative, prospective evaluation of cell damage by means of troponin I]. AB - INTRODUCTION AND OBJECTIVES: In this study we measured the concentrations of cardiac troponin I (cTnI) and several biochemical markers of myocardial damage after elective external cardioversion or internal cardioversion by specific catheters or automatic defibrillators. MATERIAL AND METHODS: Biochemical markers were analyzed prospectively for 30 consecutive patients after electrical cardioversion. Concentrations of cTnI, myoglobin, creatine kinase (CK), CK-MB and the MB/CK ratio were determined in samples before cardioversion and 2, 8 and 24 h later. The shock energy ranged from 50 to 360 joules (235 106 joules) in external cardioversions and from 3 to 37 joules (15 8 joules) in internal cardioversions. RESULTS: We detected abnormal concentrations of CK, myoglobin, CK-MB and MB/CK in 33% of the patients after external cardioversion. The concentrations of cTnI remained within normal limits at all times, with no elevations detected. Whereas no abnormal concentration of any biochemical marker was detected in any patient who required internal cardioversion for atrial fibrillation, two patients who underwent external cardioversion from an automatic defibrillator did have abnormal concentrations of CK-MB, myoglobin, and even of cTnI. CONCLUSIONS: The concentration of cTnI remained below the detection limit after external cardioversion, even though the other more non-specific markers changed. No enzyme alteration was detected in patients who underwent internal cardioversion of atrial fibrillation. PMID- 11893314 TI - [Surgery ablation of atrial fibrillation with epicardial and endocardial biauricular radiofrequency: initial experience]. AB - BACKGROUND: Atrial fibrillation is frequent in surgical patients with cardiac valvulopathies. Radiofrequency energy applied by means of surgical probes permits the reproduction of atriotomies described in the maze surgical procedure for the ablation of atrial fibrillation in a fast, safe and efficient way. This study presents our initial experience in treatment of chronic atrial fibrillation through radiofrequency performed in patients with surgical cardiac valvulopathies. PATIENTS AND METHOD: From June to November 2000, 10 patients, with surgical indications of valvulopathy, were intraoperatively treated through radiofrequency for its atrial fibrillation. Ablations were performed in the right auricle from the epicardium before starting extra corporeal circulation, and in the left auricle from the endocardium, while under circulation. Radiofrequency was applied through a surgical multielectrode probe. RESULTS: Eight patients (80%) presented some type of postoperative arrhythmia, with relapse of paroxysmal fibrillation in 3 patients and flutter in another one. At discharge, none of the patients presented relapse of chronic atrial fibrillation. There was no in hospital mortality. After a mean follow-up of 3 months (range 1-6), 8 patients (80%) have recovered and maintained sinus rhythm. Only one patient has re established echocardiographic biatrial contraction. CONCLUSIONS: Intraoperative radiofrequency has allowed us to perform the auricular lesions, in both auricles, in a simple way, with an initial effectiveness of 80%. Epicardial ablation of the right auricle was simple and safe. Although no patient presented relapse of chronic atrial fibrillation at hospital discharge, postoperative arrhythmias have continued to be the main postsurgical problem. PMID- 11893315 TI - [Cardiac troponin I in perioperative myocardial infarction after coronary artery bypass surgery]. AB - BACKGROUND AND AIMS: Myocardial infarction after coronary artery bypass grafting is a serious complication and one of the most common causes of perioperative morbidity and mortality. The present study was designed to determine the relevance of serum cardiac troponin I as a specific diagnostic marker for perioperative myocardial infarction. METHODS: A cohort of 64 patients undergoing coronary artery bypass grafting was enrolled for prospective study. Postoperative blood samples were extracted and analyzed for total creatine kinase (CK), CKMB and cardiac troponin I activity. Perioperative infarction was defined as the development of new Q waves in the postoperative electrocardiogram together with congruent regional wall motion abnormalities in the echocardiogram and CK values greater than 400 IU/l with MB fraction greater than 40 IU/l. RESULTS: Perioperative infarction occurred in 12 patients. Higher cardiac troponin I values were observed in patients experiencing perioperative myocardial infarction than in those without infarction (p < 0.001). Cardiac troponin I values higher than 12 ng/ml 10 h after release of the aortic clamp best detected the presence of perioperative myocardial infarction, with an area under the characteristic receiver operating curve of 0.91 (95% CI, 0.82-0.97), a sensitivity of 90.9%, and a specificity of 88.5%. The mean stay in the intensive care unit was significantly longer for patients who suffered perioperative myocardial infarction (6.5 8.6 days) than for patients without perioperative infarction (4.7 7.5 days) (p < 0.005). CONCLUSIONS: Cardiac troponin I elevation appears to be an early, specific marker for the diagnosis of perioperative myocardial infarction after coronary artery bypass grafting. PMID- 11893316 TI - [Clinical utilization of the coronary pressure wire]. AB - INTRODUCTION: Earlier studies have established the value of coronary pressure wires for diagnosing and monitoring the treatment of patients with coronary artery disease. In this study we demonstrated their usefulness in the daily clinical practice of a catheterization laboratory. MATERIAL AND METHODS: A retrospective study of the use of pressure wires in our laboratory between October 1998 and November 2000. The pressure wire was inserted whenever the interventional cardiologist considered it to be indicated. In all cases, pressures were recorded with a Waveguide Cardiometrics 0.014 guide (Endosonics) and hyperemia was induced by intracoronary adenosine. RESULTS: Two hundred fifty three lesions were studied in 190 patients. Indications were functional evaluation of lesions of intermediate severity for 82% (9% intrastent restenoses); guidance of balloon PTCA for 5%; and fulfillment of a research protocol for 13%. Twenty-six percent of lesions considered to be of moderate severity based on angiography were treated as a consequence of the pressures measured by the wire. A decision to begin or continue a procedure was based on wire pressures in 24% and intervention was avoided in 60%. No major complications attributable to the wire were observed. A lesion was dissected in one patient (0.5%) but it was treated without consequences. Twenty pressure wires (11%) failed to work properly during the procedure, fourteen of them (7%) before insertion. The wire could not be advanced across the lesion in one case. CONCLUSIONS: The pressure wire is useful in the daily clinical practice of a catheterization laboratory. Its most common indication is the evaluation of lesions of intermediate or unknown severity, and use is associated with few complications. PMID- 11893317 TI - [Unified three-dimensional images of myocardial perfusion and coronary angiography]. AB - INTRODUCTION AND OBJECTIVES: In everyday clinical practice, the cardiologist needs to integrate anatomical and functional information from patients with coronary artery disease. The aim of this study is to present a way to unify, in three-dimensional images, anatomical information from coronary angiography with physiological information from myocardial perfusion scintigraphy. METHODS: Three patients with one vessel disease (left anterior descending, right coronary and left circumflex arteries, respectively) scheduled for percutaneous coronary revascularization were selected. Two-dimensional angiographic images were obtained before and after revascularization. 99mTc-tetrofosmin was administered during coronary occlusion and tomographic images corresponding to the occlusion were detected after coronary dilatation. Control rest scintigraphic images were obtained after two days. The three-dimensional coronary tree from coronary angiography was superposed on the epicardial contours of the myocardial perfusion images following a method of our own. RESULTS: A correct three-dimensional reconstruction of myocardial contour and coronary tree was achieved for each patient. The three-dimensional unified images showed excellent concordance between the extent of perfusion defects and the anatomic distribution of the occluded vessel. CONCLUSIONS: Three-dimensional unification of myocardial perfusion images and coronary angiography is technically possible. This technology integrates anatomical and functional information to facilitate the cardiologist's decision-making and so improve coronary patient management. PMID- 11893318 TI - [Aortopulmonary window: clinical assessment and surgical results]. AB - INTRODUCTION AND OBJECTIVES: Aortopulmonary septal defect is an uncommon congenital cardiac anomaly. To date, approximately 300 cases have been reported. We present our experience, emphasizing the importance of early correction to avoid irreversible pulmonary hypertension. PATIENTS AND METHOD: Between 1979 and 2000, seven patients underwent surgical repair of this heart defect in our hospital. Two had type I (proximal), 4 had type II (distal) and 1 had type III (complete). Complex associated cardiac anomalies were present in 4 cases: type A interruption of the aortic arch in 2 cases, hypoplastic aortic arch in 1 and transposition of great arteries with ventricular septal defect in 1. Four cases (57%) were diagnosed by echocardiography. In all patients diagnoses were confirmed by cardiac catheterization. Patient records were reviewed retrospectively, with special attention to clinical, echocardiographic and hemodynamic data as well as surgical characteristics. RESULTS: No intraoperative deaths occurred. The patient with associated transposition of great arteries died 22 days after surgery as a result of severe pulmonary hypertension. The remaining patients are asymptomatic without treatment after a mean follow-up period of 69 months. CONCLUSIONS: Even though aortopulmonary septal defect is a rare anomaly, it should be considered whenever the course of complex congenital heart disease includes early cardiac failure and pulmonary hypertension. Repair before 6 months will prevent irreversible damage of pulmonary vessels. PMID- 11893320 TI - [Molecular genetics of cardiomyopathies]. AB - In the last decade our understanding of cardiac pathophysiology has experienced significant advances linked to major advances in molecular genetics. Although many genes are associated today with cardiac diseases, the genetics of both hypertrophic cardiomyopathy and dilated cardiomyopathy have generated great interest. The familial nature of the disease in some patients has been very useful in this regard. In addition, there are also excellent experimental models to study the implications of the genetic abnormalities. Altogether the study of the molecular genetics of the cardiomyopathies should provide not only prognostic information but also new therapeutic alternatives. PMID- 11893319 TI - [Physiologic evaluation of coronary circulation. Role of invasive and non invasive techniques]. AB - For many years, the evaluation of the extent and severity of coronary artery disease has been mainly anatomical, carried out by coronary angiography. However, this technique has methodological limitations and interobserver variability is considerable. Quantification of coronary reserve with pressure guidewires and intracoronary Doppler now provides more precise physiologic evaluation of coronary circulation. Myocardial perfusion single proton emission computed tomography and echocardiography, combined with stress and/or pharmacological challenge testing, though they are only semiquantitative techniques, also offer appropriate complements to coronary angiography in the functional evaluation of coronary patients. The aim of this paper is to discuss the clinical value of these techniques. PMID- 11893321 TI - [Swinging heart]. PMID- 11893322 TI - [Risk factors associated with endocarditis without underlying heart disease]. AB - Infective endocarditis (IE) pathogenesis has changed in the last decades and there is an increasing number of patients without predisposing heart condition. The aim of this study is to asses the clinical features of these non-drug addict patients affected with IE without underlying heart disease and to identify the potential risk factors. From 196 cases of IE, 49 (25% of the series) occurred in patients without underlying heart disease. A presumed portal of entry was identified in the majority (26 cases). The most frequent were digestive (6 cases), haemodialysis (6 cases) and central venous catheters (4 cases). Right heart valves were more often affected (29 vs 6%; p < 0.01). The distribution of the causative microorganism showed a higher proportion of Staphylococcus (57 vs 30%). Despite a similar in-hospital complication rate and a similar need of surgery during the active phase, their prognosis is better than in those with underlying heart disease. PMID- 11893323 TI - [Giant left main coronary aneurysm without associated coronary lesions]. AB - Left main coronary artery aneurysms are very infrequent (0.1%) and the majority are related to atherosclerotic obstructive lesions. Only a few isolated cases without associated coronary lesions have been reported until now. The management of these patients is not well established, thus both conservative and surgical treatments have been postulated. We present a case of giant left main coronary artery aneurysm with a diameter of 27.7 x 18.6 mm (the biggest reported until now) without coronary tree associated stenosis. It was treated conservatively with oral anticoagulation only and the 5 years evolution has been favourable. PMID- 11893324 TI - [Left ventricular free wall rupture during dobutamine stress echocardiography]. AB - Dobutamine stress echocardiography is associated with a very low rate of serious complications, lower than 0.5% (death, infarction or sustained ventricular tachycardia). We report the case of a 75 year-old female patient that suffered a fatal left ventricular free wall rupture during a dobutamine stress echocardiography after acute myocardial infarction. PMID- 11893325 TI - [Highly aggressive early prosthetic endocarditis by S. epidermidis]. AB - Prosthetic valve endocarditis is considered to be 15% of all infectious endocarditis in developed countries, more frequently during the first 45 days after surgery. Between 45 and 60% of patients with prosthetic valve endocarditis present periannular involve. The aortic valve injury and early symptoms onset after surgery are related with a higher power of aggressive prosthetic endocarditis invasion. We present the case of a patient affected with early aortic prosthetic valve endocarditis by S. epidermidis with a high aggressive and proliferating course, accompanied by fistula to left atrial, severe aortic regurgitation and left atrial roof rupture detected at the time of surgery, along with interventricular membranous septal defect. PMID- 11893326 TI - [Out-of-hospital cardiac arrest]. PMID- 11893328 TI - Microbial minimalism: genome reduction in bacterial pathogens. AB - When bacterial lineages make the transition from free-living or facultatively parasitic life cycles to permanent associations with hosts, they undergo a major loss of genes and DNA. Complete genome sequences are providing an understanding of how extreme genome reduction affects evolutionary directions and metabolic capabilities of obligate pathogens and symbionts. PMID- 11893329 TI - Opioid tolerance-in search of the holy grail. AB - Tolerance, one of several behavioral adaptations to prolonged opioid treatment, has long been explained by desensitization of opioid receptor signaling and loss of surface receptors. However, recent evidence presents an alternative hypothesis, suggesting that receptor internalization could in fact reduce tolerance in vivo. PMID- 11893330 TI - Regulatory nascent peptides in the ribosomal tunnel. AB - Accumulating evidence for nascent-peptide-mediated regulation of translation suggests that all nascent peptides do not necessarily interact with the ribosome in a similar manner. Recent studies have helped to elucidate the exit route of the nascent chain and its interactions with the ribosome. PMID- 11893331 TI - The TRP channels, a remarkably functional family. AB - TRP cation channels display an extraordinary assortment of selectivities and activation mechanisms, some of which represent previously unrecognized modes for regulating ion channels. Moreover, the biological roles of TRP channels appear to be equally diverse and range from roles in pain perception to male aggression. PMID- 11893332 TI - Structural organization of bacterial RNA polymerase holoenzyme and the RNA polymerase-promoter open complex. AB - We have used systematic fluorescence resonance energy transfer and distance constrained docking to define the three-dimensional structures of bacterial RNA polymerase holoenzyme and the bacterial RNA polymerase-promoter open complex in solution. The structures provide a framework for understanding sigma(70)-(RNA polymerase core), sigma(70)-DNA, and sigma(70)-RNA interactions. The positions of sigma(70) regions 1.2, 2, 3, and 4 are similar in holoenzyme and open complex. In contrast, the position of sigma(70) region 1.1 differs dramatically in holoenzyme and open complex. In holoenzyme, region 1.1 is located within the active-center cleft, apparently serving as a "molecular mimic" of DNA, but, in open complex, region 1.1 is located outside the active center cleft. The approach described here should be applicable to the analysis of other nanometer-scale complexes. PMID- 11893333 TI - A regulated two-step mechanism of TBP binding to DNA: a solvent-exposed surface of TBP inhibits TATA box recognition. AB - The TATA box binding protein TBP plays a universally important role in eukaryotic nuclear transcription. By mutagenesis, we have discovered a solvent-exposed surface of the structured TBP core domain that is important for inhibition of the DNA binding and DNA-bending activities of full-length wild-type TBP. Full-length wild-type TBP initially binds the TATA box to form an unstable complex containing unbent DNA, and then it slowly forms a stable complex containing bent DNA. TFIIB greatly accelerates formation of a bent TFIIB-TBP-TATA box complex, and the inhibitory DNA binding surface of TBP contributes to the cooperativity of binding to TFIIB. Using TBP and TFIIB, we show that TBP can bind the TATA box through a regulated two-step mechanism, involving a transition from unbent complex to bent complex. PMID- 11893334 TI - The ribosomal exit tunnel functions as a discriminating gate. AB - Translation of SecM stalls unless its N-terminal part is "pulled" by the protein export machinery. Here we show that the sequence motif FXXXXWIXXXXGIRAGP that includes a specific arrest point (Pro) causes elongation arrest within the ribosome. Mutations that bypass the elongation arrest were isolated in 23S rRNA and L22 r protein. Such suppressor mutations occurred at a few specific residues of these components, which all face the narrowest constriction of the ribosomal exit tunnel. Thus, we suggest that this region of the exit tunnel interacts with nascent translation products and functions as a discriminating gate. PMID- 11893335 TI - Trypanosome mitochondrial 3' terminal uridylyl transferase (TUTase): the key enzyme in U-insertion/deletion RNA editing. AB - A 3' terminal RNA uridylyltransferase was purified from mitochondria of Leishmania tarentolae and the gene cloned and expressed from this species and from Trypanosoma brucei. The enzyme is specific for 3' U-addition in the presence of Mg(2+). TUTase is present in vivo in at least two stable configurations: one contains a approximately 500 kDa TUTase oligomer and the other a approximately 700 kDa TUTase complex. Anti-TUTase antiserum specifically coprecipitates a small portion of the p45 and p50 RNA ligases and approximately 40% of the guide RNAs. Inhibition of TUTase expression in procyclic T. brucei by RNAi downregulates RNA editing and appears to affect parasite viability. PMID- 11893336 TI - A Ca(2+) switch aligns the active site of calpain. AB - Ca(2+) signaling by calpains leads to controlled proteolysis during processes ranging from cytoskeleton remodeling in mammals to sex determination in nematodes. Deregulated Ca(2+) levels result in aberrant proteolysis by calpains, which contributes to tissue damage in heart and brain ischemias as well as neurodegeneration in Alzheimer's disease. Here we show that activation of the protease core of mu calpain requires cooperative binding of two Ca(2+) atoms at two non-EF-hand sites revealed in the 2.1 A crystal structure. Conservation of the Ca(2+) binding residues defines an ancestral general mechanism of activation for most calpain isoforms, including some that lack EF-hand domains. The protease region is not affected by the endogenous inhibitor, calpastatin, and may contribute to calpain-mediated pathologies when the core is released by autoproteolysis. PMID- 11893337 TI - AtPIN4 mediates sink-driven auxin gradients and root patterning in Arabidopsis. AB - In contrast to animals, little is known about pattern formation in plants. Physiological and genetic data suggest the involvement of the phytohormone auxin in this process. Here, we characterize a novel member of the PIN family of putative auxin efflux carriers, Arabidopsis PIN4, that is localized in developing and mature root meristems. Atpin4 mutants are defective in establishment and maintenance of endogenous auxin gradients, fail to canalize externally applied auxin, and display various patterning defects in both embryonic and seedling roots. We propose a role for AtPIN4 in generating a sink for auxin below the quiescent center of the root meristem that is essential for auxin distribution and patterning. PMID- 11893338 TI - Regulation of Frizzled by fat-like cadherins during planar polarity signaling in the Drosophila compound eye. AB - Planar polarity is evident in the coordinated orientation of ommatidia in the Drosophila eye. This process requires that the R3 photoreceptor precursor of each ommatidium have a higher level of Frizzled signaling than its neighboring R4 precursor. We show that two cadherin superfamily members, Fat and Dachsous, and the transmembrane/secreted protein Four-jointed play important roles in this process. Our data support a model in which the bias of Frizzled signaling between the R3/R4 precursors results from higher Fat function in the precursor cell closer to the equator, which becomes R3. We also provide evidence that positional information regulating Fat action is provided by graded expression of Dachsous across the eye and the action of Four-jointed, which is expressed in an opposing expression gradient and appears to modulate Dachsous function. PMID- 11893339 TI - Expression of constitutively active CREB protein facilitates the late phase of long-term potentiation by enhancing synaptic capture. AB - Restricted and regulated expression in mice of VP16-CREB, a constitutively active form of CREB, in hippocampal CA1 neurons lowers the threshold for eliciting a persistent late phase of long-term potentiation (L-LTP) in the Schaffer collateral pathway. This L-LTP has unusual properties in that its induction is not dependent on transcription. Pharmacological and two-pathway experiments suggest a model in which VP16-CREB activates the transcription of CRE-driven genes and leads to a cell-wide distribution of proteins that prime the synapses for subsequent synapse-specific capture of L-LTP by a weak stimulus. Our analysis indicates that synaptic capture of CRE-driven gene products may be sufficient for consolidation of LTP and provides insight into the molecular mechanisms of synaptic tagging and synapse-specific potentiation. PMID- 11893340 TI - A TRP channel that senses cold stimuli and menthol. AB - A distinct subset of sensory neurons are thought to directly sense changes in thermal energy through their termini in the skin. Very little is known about the molecules that mediate thermoreception by these neurons. Vanilloid Receptor 1 (VR1), a member of the TRP family of channels, is activated by noxious heat. Here we describe the cloning and characterization of TRPM8, a distant relative of VR1. TRPM8 is specifically expressed in a subset of pain- and temperature-sensing neurons. Cells overexpressing the TRPM8 channel can be activated by cold temperatures and by a cooling agent, menthol. Our identification of a cold sensing TRP channel in a distinct subpopulation of sensory neurons implicates an expanded role for this family of ion channels in somatic sensory detection. PMID- 11893341 TI - Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. AB - The first structure of a flavivirus has been determined by using a combination of cryoelectron microscopy and fitting of the known structure of glycoprotein E into the electron density map. The virus core, within a lipid bilayer, has a less ordered structure than the external, icosahedral scaffold of 90 glycoprotein E dimers. The three E monomers per icosahedral asymmetric unit do not have quasiequivalent symmetric environments. Difference maps indicate the location of the small membrane protein M relative to the overlaying scaffold of E dimers. The structure suggests that flaviviruses, and by analogy also alphaviruses, employ a fusion mechanism in which the distal beta barrels of domain II of the glycoprotein E are inserted into the cellular membrane. PMID- 11893343 TI - The relation between managed care market share and the treatment of elderly fee for-service patients with myocardial infarction. AB - PURPOSE: To determine if greater managed care market share is associated with greater use of recommended therapies for fee-for-service patients with acute myocardial infarction. SUBJECTS AND METHODS: We examined the care of 112,900 fee for-service Medicare beneficiaries aged > or = 65 years who resided in one of 320 metropolitan statistical areas and who were admitted with an acute myocardial infarction between February 1994 through July 1995. Use of recommended medical treatments and 30-day survival were determined for areas with low (<10%), medium (10% to 30%), and high (>30%) managed care market share. RESULTS: After adjustment for severity of illness, teaching status of the admission hospital, and area characteristics, areas with high levels of managed care had greater use of beta-blockers (relative risk [RR] for greater use = 1.18; 95% confidence interval [CI]: 1.06 to 1.29) and aspirin at discharge (RR = 1.05; 95% CI: 1.02 to 1.07), but less appropriate coronary angiography (RR = 0.93; 95% CI: 0.86 to 1.01) and reperfusion (RR = 0.95; 95% CI: 0.85 to 1.03) when compared with areas with low levels of managed care. CONCLUSIONS: Medicare beneficiaries with fee-for service insurance who resided in areas with high managed care activity were more likely to have received appropriate treatment with beta-blockers and aspirin, and less likely to have undergone coronary angiography following admission for myocardial infarction. Thus, the effects of managed care may not be limited to managed care enrollees. PMID- 11893342 TI - Potential cost savings of erythropoietin administration in end-stage renal disease. AB - BACKGROUND: In a Department of Veterans Affairs randomized controlled trial, a lower dose of recombinant human erythropoietin (epoetin) was shown to attain target hematocrit levels when administered subcutaneously compared with intravenously. Since epoetin is expensive, optimizing the therapeutic effect of epoetin using a strategy that includes subcutaneous administration could lead to substantial cost savings. METHODS: We used an economic cost projection model to estimate potential savings to the Medicare End-Stage Renal Disease Program that could occur during a transition from intravenous to subcutaneous administration of epoetin among hemodialysis patients. Data included clinical results from the Department of Veterans Affairs randomized controlled trial, the 1998 Centers for Medicare and Medicaid Services' End-Stage Renal Disease Core Indicators Survey, and the 1997-1998 Medicare claims files. In sensitivity analyses, we varied the expected dose reductions (10% to 50%) and the proportion of patients (25% to 100%) who switched to subcutaneous administration. RESULTS: Medicare cost savings were estimated at $47 to $142 million annually as 25% to 75% of hemodialysis patients who received epoetin intravenously switched to subcutaneous administration while reducing the dose by 32%. A minimal reduction (10%) in epoetin dose would result in Medicare cost savings of an estimated $15 to $44 million annually. CONCLUSION: Administering epoetin subcutaneously would provide substantial cost savings to Medicare. For the transition to occur, consensus among stakeholders is needed, especially among patients whose treatment satisfaction and health-related quality of life would be most affected. PMID- 11893344 TI - Clinical presentation of genetically defined patients with hypokalemic salt losing tubulopathies. AB - PURPOSE: Hypokalemic salt-losing tubulopathies (Bartter-like syndromes) comprise a set of clinically and genetically distinct inherited renal disorders. Mutations in four renal membrane proteins involved in electrolyte reabsorption have been identified in these disorders: the furosemide-sensitive sodium-potassium-chloride cotransporter NKCC2, the potassium channel ROMK, the chloride channel ClC-Kb, and the thiazide-sensitive sodium-chloride cotransporter NCCT. The aim of this study was to characterize the clinical features associated with each mutation in a large cohort of genetically defined patients. PATIENTS AND METHODS: The phenotypic characteristics of 65 patients with molecular defects in NKCC2, ROMK, ClC-Kb, or NCCT were collected retrospectively. RESULTS: ROMK and NKCC2 patients presented with polyhydramnios, nephrocalcinosis, and hypo- or isosthenuria. Hypokalemia was less severe in the ROMK patients compared with the NKCC2 patients. In contrast, NCCT patients had hypocalciuria, hypomagnesemia, and marked hypokalemia. While this dissociation of renal calcium and magnesium handling was also observed in some ClC-Kb patients, a few ClC-Kb patients presented with hypercalciuria and hypo- or isosthenuria. CONCLUSIONS: ROMK, NKCC2, and NCCT mutations usually have uniform clinical presentations, whereas mutations in ClC-Kb occasionally lead to phenotypic overlaps with the NCCT or, less commonly, with the ROMK/NKCC2 cohort. Based on these results, we propose an algorithm for the molecular diagnosis of hypokalemic salt-losing tubulopathies. PMID- 11893345 TI - A randomized, placebo-controlled, double-blind, flexible-dose study of fluoxetine in the treatment of women with fibromyalgia. AB - PURPOSE: To assess the efficacy of fluoxetine in the treatment of patients with fibromyalgia. SUBJECTS AND METHODS: Sixty outpatients (all women, aged 21-71 years) with fibromyalgia were randomly assigned to receive fluoxetine (10-80 mg/d) or placebo for 12 weeks in a double-blind, parallel-group, flexible-dose study. The primary outcome measures were the Fibromyalgia Impact Questionnaire total score (score range, 0 [no impact] to 80) and pain score (score range, 0 10). Secondary measures included the McGill Pain Questionnaire, change in the number of tender points, and total myalgic score. RESULTS: In the intent-to-treat analysis, women who received fluoxetine (mean [+/- SD] dose, 45 +/- 25 mg/d) had significant (P = 0.005) improvement in the Fibromyalgia Impact Questionnaire total score compared with those who received placebo, with a difference of -12 (95% confidence interval [CI]: -19 to -4). They also had significant (P = 0.002) improvement in the Fibromyalgia Impact Questionnaire pain score (difference, -2.2 [95% CI: -3.6 to -0.9]), as well as in the Fibromyalgia Impact Questionnaire fatigue (P = 0.05) and depression (P = 0.01) scores and the McGill Pain Questionnaire (P = 0.01), when compared with subjects who received placebo. Although counts for the number of tender points and total myalgic scores improved more in the fluoxetine group than in the placebo group, these differences were not statistically significant. CONCLUSIONS: In a 12-week, flexible-dose, placebo controlled trial, fluoxetine was found to be effective on most outcome measures and generally well tolerated in women with fibromyalgia. PMID- 11893346 TI - Clinical prediction of deep venous thrombosis using two risk assessment methods in combination with rapid quantitative D-dimer testing. AB - PURPOSE: The optimal approach to diagnosing deep venous thrombosis is not entirely clear. In this prospective cohort study, we aimed to evaluate the yield of two methods of assessing the pretest probability of deep venous thrombosis-the treating physician's implicit assessment and the Wells score, a validated prediction rule that incorporates signs, symptoms, and the presence or absence of an alternative diagnosis-used in isolation and in combination with D-dimer measurement. SUBJECTS AND METHODS: We studied 278 patients who were referred for suspicion of deep venous thrombosis. All patients were stratified into groups of low, moderate, or high risk of deep venous thrombosis on the basis of the clinical assessment and Wells score, and underwent rapid quantitative D-dimer testing (with a cutoff of 500 microg/mL), ultrasound examination, and follow-up for the occurrence of venous thromboembolism. RESULTS: Eighty-two patients (29%) had a deep venous thrombosis. The accuracy of both methods was good (area under the receiver operating characteristic curve = 0.72), despite only fair agreement at the level of individual patients (weighted kappa = 0.31; 95% confidence interval [CI]: 0.23 to 0.40). The negative predictive value of D-dimer measurement was 96% (95% CI: 91% to 100%). When restricted to patients with low pretest probability, the negative predictive value of D-dimer measurement was 100% (95% CI: 96% to 100%) with the use of the Wells score and 96% (95% CI: 88% to 100%) with the physician's assessment. Our results were unchanged in analyses restricted to patients with proximal deep venous thrombosis or outpatients. CONCLUSION: Clinical assessment to stratify a patient's likelihood of having deep venous thrombosis should be taught to physicians. PMID- 11893347 TI - Antimicrobial therapy of gram-negative bacteremia at two university-affiliated medical centers. AB - PURPOSE: To describe antimicrobial prescribing practices and patient outcomes associated with the treatment of aerobic gram-negative rod bacteremia at two university-affiliated medical centers. SUBJECTS AND METHODS: All adult patients with gram-negative bacteremia (N = 326) who were at Stanford and University of California, San Francisco (UCSF) Hospitals from September 1, 1996 through August 31, 1997 were evaluated via retrospective review of medical records. RESULTS: Most patient characteristics were similar between institutions; however, patients at Stanford were more likely to have had a diagnosis of bone marrow transplantation, liver failure, or poor nutritional status, while more patients at UCSF had solid organ transplant, diabetes, pulmonary disease, or hypotension. The bacteriology was similar at both sites, with Escherichia coli the predominant pathogen (139 [43%] of 326). The majority of episodes were community acquired (67% [218/326]). Patients at Stanford were more likely to have been treated empirically with aminoglycosides (28% vs. 7%, P <0.001) and noncephalosporin beta lactams (31% vs. 11%, P <0.001), while patients at UCSF were more likely to have received cephalosporins (62% vs. 29%, P <0.001) and fluoroquinolones (21% vs. 11%, P = 0.02). These patterns continued for definitive therapy. Overall mortality was 60 (19%) of 326. Several risk factors were associated with 14-day mortality, including severity of illness, neutropenia, diabetes mellitus, use of vasopressors, and empiric use of a noncephalosporin beta-lactam. CONCLUSION: Prescribing practices for the treatment of gram-negative bacteremia differed significantly in the two institutions despite similar patients and pathogens. PMID- 11893348 TI - Improving oral presentation skills with a clinical reasoning curriculum: a prospective controlled study. AB - PURPOSE: The oral case presentation is an essential part of clinical medicine, but teaching medical students to present clinical data remains difficult. Presentation skills depend on the ability to obtain, process, and organize patient data. Clinical reasoning is fundamental to the development of these skills. We compared a clinical reasoning curriculum with standard ward instruction for improving presentation skills and clinical performance. SUBJECTS AND METHODS: Between October 1998 and May 1999, 62 third-year medical students at three hospitals were assigned to a 4-week clinical reasoning curriculum (n = 27) or a control group (n = 35) that underwent routine instruction. The curriculum consisted of four 1-hour group sessions and 1 hour of individual videotaped instruction, and taught students to use the principles of clinical reasoning, such as generation and refinement of diagnostic hypothesis, interpretation of diagnostic tests, and causal reasoning, to determine data for inclusion in the oral presentation. We videotaped students presenting two standardized case histories; one at baseline and a second 4 weeks later. Two independent evaluators who were blinded to the group assignments reviewed the videotapes and scored them for presentation quality and efficiency, and general speaking ability. RESULTS: Mean (+/- SD) presentation times at baseline were similar in the two groups (intervention group: 8 +/- 2 minutes; control group: 8 +/- 2 minutes; P = 0.74). Presentation time in students who were taught clinical reasoning decreased by 3 +/- 2 minutes, but increased by 2 +/- 2 minutes in control students. The difference in the changes between the groups was statistically significant (mean difference = 4 minutes; 95% confidence interval [CI]: 3 to 5 minutes; P <0.001). Presentation quality scores at baseline were similar in both groups (intervention group: 17 +/- 8 points; control group: 20 +/- 7 points; P = 0.11). Students who were taught the clinical reasoning curriculum had an improvement of 9 +/- 6 points in the quality of their presentations, while control students had an improvement of 2 +/- 7 points (on a scale of 4-36). The difference in the changes between the groups was statistically significant (mean difference = 4 points; 95% CI: 1 to 7 points; P = 0.04). CONCLUSION: A clinical reasoning curriculum, in combination with video-based individual instruction, improves the efficiency and quality of oral presentations, and may augment clinical performance. PMID- 11893349 TI - Predicting pulmonary complications after nonthoracic surgery: a systematic review of blinded studies. AB - PURPOSE: To determine the performance of variables commonly used in the prediction of postoperative pulmonary complications in patients undergoing nonthoracic surgery. METHODS: We conducted a systematic review of the literature in English, using MEDLINE (1966-2001), manual searches of identified articles, and contact with content experts. All studies reporting independent and blinded comparisons of preoperative or operative factors with postoperative pulmonary complications were included. Two reviewers independently abstracted inclusion and exclusion criteria, study designs, patient characteristics, predictors of interest, and the nature and occurrence of postoperative pulmonary complications. RESULTS: Seven studies fulfilled the inclusion criteria. The definition of postoperative pulmonary complications differed among studies, and the incidence of postoperative pulmonary complications varied from 2% to 19%. Of the 28 preoperative or operative predictors that were evaluated in the 7 studies, 16 were associated significantly with postoperative pulmonary complications, although only 2 (duration of anesthesia and postoperative nasogastric tube placement) were significant in more than one study. The positive (2.2 to 5.1) and negative (0.2 to 0.8) likelihood ratios for these 16 variables suggest that they have only modest predictive value. Neither hypercarbia nor reduced spirometry values were independently associated with an increased risk of postoperative pulmonary complications. CONCLUSION: Few studies have rigorously evaluated the performance of the preoperative or operative variables in the prediction of postoperative pulmonary complications. Prospective studies with independent and blinded comparisons of these variables with postoperative outcomes are needed. PMID- 11893350 TI - Rapid suppression of alcohol withdrawal syndrome by baclofen. PMID- 11893351 TI - Squeezing more cost and care out of dialysis: our patients would pay the price. PMID- 11893352 TI - Genetic forms of renal potassium and magnesium wasting. PMID- 11893353 TI - Treatment trials of unexplained symptom syndromes: time for larger and longer trials to address the complexities. PMID- 11893354 TI - It's in the air: managed care and spillover effects. PMID- 11893355 TI - Atypical hemolytic uremic syndrome caused by infection with a non-Shiga toxin producing strain of Escherichia Coli. PMID- 11893356 TI - The new era in managing type 2 diabetes. PMID- 11893357 TI - Nonconvulsive status epilepticus. PMID- 11893359 TI - Hyporeninemic hypoaldosteronism associated with Sjogren's syndrome. PMID- 11893360 TI - Valsalva retinopathy and optic nerve drusen in a patient with cystic fibrosis. PMID- 11893361 TI - Hypoalbuminemia as a risk factor for over-anticoagulation. PMID- 11893362 TI - Integrating geriatrics and subspecialty internal medicine: results of a survey on patient care practices, training, attitudes, and research. PMID- 11893363 TI - Hospital outcomes in major teaching, minor teaching, and nonteaching hospitals in New York state. AB - PURPOSE: The possible benefit that hospital teaching status may confer in the care of patients with cardiovascular disease is unknown. Our purpose was to determine the effect of hospital teaching status on in-hospital mortality, use of invasive procedures, length of stay, and charges in patients with myocardial infarction, heart failure, or stroke. SUBJECTS AND METHODS: We analyzed a New York State hospital administrative database containing information on 388 964 consecutive patients who had been admitted with heart failure (n = 173 799), myocardial infarction (n = 121 209), or stroke (n = 93 956) from 1993 to 1995. We classified the 248 participating acute care hospitals by teaching status (major, minor, nonteaching). The primary outcomes were standardized in-hospital mortality ratios, defined as the ratio of observed to predicted mortality. RESULTS: Standardized in-hospital mortality ratios were significantly lower in major teaching hospitals (0.976 for heart failure, 0.945 for myocardial infarction, 0.958 for stroke) than in nonteaching hospitals (1.01 for heart failure, 1.01 for myocardial infarction, 0.995 for stroke). Standardized in-hospital mortality ratios were significantly higher for patients with stroke (1.06) but not heart failure (1.0) or myocardial infarction (1.06) in minor teaching hospitals than in nonteaching hospitals. Compared with nonteaching hospitals, use of invasive cardiac procedures and adjusted hospital charges were significantly greater in major and minor teaching hospitals for all three conditions. The adjusted length of stay was also shorter for myocardial infarction in major teaching hospitals and longer for stroke in minor teaching hospitals. CONCLUSION: Major teaching hospital status was an important determinant of outcomes in patients hospitalized with myocardial infarction, heart failure, or stroke in New York State. PMID- 11893365 TI - Beta-blockers are associated with lower C-reactive protein concentrations in patients with coronary artery disease. AB - PURPOSE: C-reactive protein is an important risk factor for coronary artery disease, and plasma concentrations are lowered by treatment with pravastatin and aspirin. We examined whether other cardiovascular drugs that are used in the treatment of ischemic heart disease affect C-reactive protein concentrations. SUBJECTS AND METHODS: Plasma C-reactive protein concentration was measured by high sensitivity immunonephelometric assay in 333 consecutive patients with stable angina and confirmed coronary artery disease who underwent diagnostic angiography. RESULTS: Patients prescribed beta-blockers had significantly lower mean C-reactive protein concentrations than did patients in whom these were not prescribed (by 1.2 mg/L, or 40% difference in geometric mean concentration; P <0.001). This association remained significant (P = 0.03) after excluding patients with contraindications to the use of beta-blockers, and adjusting for the probability of beta-blocker therapy (propensity score) and other clinical predictors of C-reactive protein concentration, including body mass index, high density lipoprotein cholesterol level, family history of coronary artery disease, and angiographic severity. No differences among types or dosages of beta-blockers were evident. CONCLUSION: Beta-blockers may affect C-reactive protein concentrations. Randomized studies are required to confirm these findings. PMID- 11893364 TI - Fluorine-18 fluorodeoxyglucose positron emission tomography, gallium-67 scintigraphy, and conventional staging for Hodgkin's disease and non-Hodgkin's lymphoma. AB - PURPOSE: To compare fluorine-18 fluorodeoxyglucose positron emission tomography (PET) and gallium scanning with each other and with conventional staging, for patients with Hodgkin's disease or non-Hodgkin's lymphoma. SUBJECTS AND METHODS: Fifty patients had PET, gallium scanning, and conventional staging of newly diagnosed or progressive Hodgkin's disease or non-Hodgkin's lymphoma. Disease sites, stage, and treatment plans were assessed retrospectively. RESULTS: Positron emission tomography and gallium scanning each upstaged 14% of patients (n = 7). Management was altered by PET in 9 cases (18%) and by gallium scanning in 7 (14%, P = 0.6). Disease was evident in 117 sites in 42 patients. The case positivity rate for conventional assessment was 90%; for PET, 95%; for gallium scanning, 88%; for conventional assessment plus PET, 100%; and for conventional assessment plus gallium scanning, 98%. Site positivity rates for conventional assessment were 68%; for PET, 82%; for gallium scanning, 69% (conventional vs. PET, P = 0.01; conventional vs. gallium scanning, P = 0.9; PET vs. gallium scanning, P = 0.01); for conventional assessment plus PET, 96%; and for conventional assessment plus gallium scanning, 94%. Positron emission tomography and gallium scanning were entirely concordant in 31 patients; in the other 19 patients, PET identified 25 sites missed by gallium scanning, whereas gallium scanning identified 10 sites missed by PET. CONCLUSION: In this retrospective study, PET demonstrated a higher site positivity rate than did gallium scanning, with similar case positivity rates. These data support the use of PET in place of gallium scanning for the staging of patients with Hodgkin's disease or non Hodgkin's lymphoma. PMID- 11893366 TI - Severe hypertriglyceridemia with insulin resistance is associated with systemic inflammation: reversal with bezafibrate therapy in a randomized controlled trial. AB - PURPOSE: To determine whether hypertriglyceridemia is associated with systemic inflammation, which may contribute to the increased cardiovascular risk in patients who have hypertriglyceridemia. In addition, we investigated whether fibrates reverse this inflammatory state. PATIENTS AND METHODS: Serum lipid levels, body mass index, insulin resistance, and inflammatory parameters were compared between 18 patients who had severe hypertriglyceridemia without cardiovascular disease and 20 normolipidemic controls. We measured the ex vivo production capacity of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 after whole-blood stimulation with lipopolysaccharide, as well as circulating levels of C-reactive protein and fibrinogen. A randomized controlled trial was conducted to determine whether bezafibrate (400 mg administered daily for 6 weeks) affected these parameters in hypertriglyceridemic patients. RESULTS: When compared with normolipidemic controls, hypertriglyceridemic patients had significantly lower high-density lipoprotein (HDL) cholesterol and higher triglyceride levels, body mass index, and insulin resistance. In addition, hypertriglyceridemic patients had a significantly higher production capacity of TNF-alpha (mean difference, 11 700 pg/mL; 95% confidence interval [CI]: 7800 to 15,700 pg/mL]) and IL-6 (mean difference, 20,400 pg/mL; 95% CI: 7800 to 32,900 pg/mL), and higher levels of C-reactive protein (mean difference, 0.8 mg/L; 95% CI: 0.1 to 2.4 mg/L) and fibrinogen (mean difference, 0.8 g/dL; 95% CI: 0.3 to 1.3 g/dL). Bezafibrate therapy significantly increased HDL cholesterol levels, reduced triglyceride and insulin resistance levels, and reduced production capacity of TNF-alpha and IL-6, as well as levels of C-reactive protein and fibrinogen. CONCLUSION: Systemic inflammation is present in patients who have the clinical phenotype that is associated with severe hypertriglyceridemia, and may contribute to the increased risk of cardiovascular disease in these patients. Bezafibrate has anti-inflammatory effects in these patients. PMID- 11893367 TI - Improvement in spine bone density and reduction in risk of vertebral fractures during treatment with antiresorptive drugs. AB - PURPOSE: To estimate how much the improvement in bone mass accounts for the reduction in risk of vertebral fracture that has been observed in randomized trials of antiresorptive treatments for osteoporosis. METHODS: After a systematic search, we conducted a meta-analysis of 12 trials to describe the relation between improvement in spine bone mineral density and reduction in risk of vertebral fracture in postmenopausal women. We also used logistic models to estimate the proportion of the reduction in risk of vertebral fracture observed with alendronate in the Fracture Intervention Trial that was due to improvement in bone mineral density. RESULTS: Across the 12 trials, a 1% improvement in spine bone mineral density was associated with a 0.03 decrease (95% confidence interval [CI]: 0.02 to 0.05) in the relative risk (RR) of vertebral fracture. The reductions in risk were greater than predicted from improvement in bone mineral density; for example, the model estimated that treatments predicted to reduce fracture risk by 20% (RR = 0.80), based on improvement in bone mineral density, actually reduce the risk of fracture by about 45% (RR = 0.55). In the Fracture Intervention Trial, improvement in spine bone mineral density explained 16% (95% CI: 11% to 27%) of the reduction in the risk of vertebral fracture with alendronate. CONCLUSION: Improvement in spine bone mineral density during treatment with antiresorptive drugs accounts for a predictable but small part of the observed reduction in the risk of vertebral fracture. PMID- 11893368 TI - Effects of adjustment for referral bias on the sensitivity and specificity of single photon emission computed tomography for the diagnosis of coronary artery disease. AB - PURPOSE: Referral bias, in which the result of a diagnostic test affects the subsequent referral for a more definitive test, influences the accuracy of noninvasive tests for coronary artery disease. This study evaluates the effect of referral bias on the apparent accuracy of single photon emission computed tomography (SPECT). METHODS: Over a 10-year period, 14,273 patients without known coronary artery disease underwent stress SPECT. Coronary angiography was performed within 3 months after the stress test in 1853 patients (13%). The apparent sensitivity, specificity, and likelihood ratios of SPECT were determined in these patients, and then adjusted for referral bias using two different formulas. RESULTS: The overwhelming majority (95%) of patients who underwent angiography had abnormal SPECT images. Apparent values for test indices were a sensitivity of 98%, a specificity of 13%, a likelihood ratio for a positive test of 1.1, and a likelihood ratio for a negative test of 0.15. Test indices adjusted for referral bias (using the two methods) were a sensitivity of 65% or 67%, a specificity of 67% or 75%, a likelihood ratio for a positive test of 2.0 or 2.7, and a likelihood ratio for a negative test of 0.44 or 0.52. CONCLUSION: Referral bias has a marked effect on the apparent accuracy of stress SPECT for the diagnosis of coronary disease. Adjustment for referral bias yields estimates for sensitivity and specificity and likelihood ratios that better reflect the accuracy of the technique. PMID- 11893369 TI - N-3 polyunsaturated fatty acids in coronary heart disease: a meta-analysis of randomized controlled trials. AB - PURPOSE: Observational studies have shown an inconsistent association between n-3 polyunsaturated fatty acids and the risk of coronary heart disease. We investigated the effects of dietary and non-dietary (supplemental) intake of n-3 polyunsaturated fatty acids on coronary heart disease. SUBJECTS AND METHODS: We searched the literature to identify randomized controlled trials that compared dietary or non-dietary intake of n-3 polyunsaturated fatty acids with a control diet or placebo in patients with coronary heart disease. Studies had to have at least 6 months of follow-up data, and to have reported clinical endpoint data. We identified 11 trials, published between 1966 and 1999, which included 7951 patients in the intervention and 7855 patients in the control groups. RESULTS: The risk ratio of nonfatal myocardial infarction in patients who were on n-3 polyunsaturated fatty acid-enriched diets compared with control diets or placebo was 0.8 (95% confidence interval [CI]: 0.5 to 1.2, P = 0.16; Breslow-Day test for heterogeneity, P = 0.01), and the risk ratio of fatal myocardial infarction was 0.7 (95% CI: 0.6 to 0.8, P <0.001; heterogeneity P >0.20). In 5 trials, sudden death was associated with a risk ratio of 0.7 (95% CI: 0.6 to 0.9, P <0.01; heterogeneity P >0.20), whereas the risk ratio of overall mortality was 0.8 (95% CI: 0.7 to 0.9, P <0.001; heterogeneity P >0.20). There was no difference in summary estimates between dietary and non-dietary interventions of n-3 polyunsaturated fatty acids for all endpoints. CONCLUSION: This meta-analysis suggests that dietary and non-dietary intake of n-3 polyunsaturated fatty acids reduces overall mortality, mortality due to myocardial infarction, and sudden death in patients with coronary heart disease. PMID- 11893370 TI - Metabolic causes and prevention of ventricular fibrillation during acute coronary syndromes. AB - The mechanisms leading to ventricular fibrillation that occur during acute myocardial ischemia are ill understood. Whether primary ventricular fibrillation is due to a transient imbalance of electrolytes, an alteration of membrane permeability, electrical re-entry phenomena, or other factors, one overriding influence is the development of regional myocardial energy crises. Acute alteration in the balance of substrate supply may lead, during greatly reduced blood flow, to instability of myocardial electrical conduction with the development of re-entry circuits. An immediate response to the angor animi and initial symptoms of an acute coronary syndrome is a rapid and marked increase in catecholamine release, which leads to adipose tissue lipolysis with an acute increase in plasma free fatty acid concentrations, suppression of insulin activity, and a reduction in glucose uptake by the myocardium. The utilization of free fatty acids instead of glucose by the ischemic myocardium could precipitate regional oxygen or energy crises. Prevention therefore should focus on minimizing the catecholamine response and providing the myocardium with an optimum supply of energy substrates. Since catecholamines are inotropic, the aim should be to redress the imbalance of substrate availability by controlling adipose lipolysis with reduction of plasma free fatty acid concentrations, increasing the availability of glucose, or both. Other approaches include inhibition of acylcarnitine transport and manipulation of fatty acid intermediaries. To combat primary ventricular fibrillation, preventive treatment must be established within 6 to 10 hours of the onset of ischemia. There is already experimental and clinical evidence that antilipolytic drugs decrease the incidence of ventricular fibrillation, but their potential has not been explored extensively. PMID- 11893371 TI - Cases from the Osler Medical Service at Johns Hopkins University. Hodgkin's disease with Pel-Ebstein fevers. PMID- 11893372 TI - Assessing hospital outcomes: it's time to get inside the black box. PMID- 11893373 TI - Fish and N-3 fatty acids for the prevention and treatment of coronary heart disease: nutrition is not pharmacology. PMID- 11893374 TI - Fluorine-18 fluorodeoxyglucose positron emission tomography for lymphoma: incorporating new technology into clinical care. PMID- 11893375 TI - Adjusting for bias in diagnostic reports. PMID- 11893376 TI - Aeromonas jandaei cellulitis and bacteremia in a man with diabetes. PMID- 11893377 TI - Postinfantile giant cell hepatitis associated with long-term elevated transaminase levels in treated Graves' disease. PMID- 11893378 TI - Anorexia nervosa and the risk of sudden death. PMID- 11893379 TI - Severe Clostridium difficile colitis: the role of intracolonic vancomycin? PMID- 11893380 TI - Atorvastatin-induced reversible positive antinuclear antibodies. PMID- 11893381 TI - Hepatitis B virus-associated aplastic anemia followed by myelodysplastic syndrome. PMID- 11893382 TI - Control of life-threatening pulmonary bleeding with activated recombinant factor VII. PMID- 11893383 TI - Inhaled fluticasone and zafirlukast in persistent asthma. PMID- 11893385 TI - Drug-induced hyperkalemia. PMID- 11893387 TI - Mentoring in medicine: keys to satisfaction. PMID- 11893388 TI - Immunomodulatory activity of acidic xylans in relation to their structural and molecular properties. AB - The structure/function relationship of two acidic heteroxylan types, the arabino (glucurono)xylan from corn cobs (AGX) and 4-O-methylglucuronoxylans (GXs) from beechwood and three medicinal herbs (Rudbeckia, Altheae, and Mahonia), has been studied. The effect of the molecular mass of AGX, as well as the content and distribution of the 4-O-methylglucuronic acid side chains in GXs on the immunological activity of these xylans was characterized by their biological response in the mitogenic and comitogenic thymocyte in vitro tests. Depolymerization of AGX by ultrasonication resulted in unequivocal decrease of the immunomodulatory activity, whereas already a short treatment by endo-beta-1,4 xylanase brought about a significant increase in its activity when applied in the highest dose. In the case of the GX samples, neither the uronic acid content nor the distribution pattern of the uronic acid side chains was found to be determinant for the expression of their immunomodulatory activity. PMID- 11893389 TI - A molecular description of the gelation mechanism of curdlan. AB - The gelation of aqueous suspensions of the polysaccharide curdlan has been studied by dynamic rheological measurements, differential scanning calorimetry, and low-resolution time-domain 1H-NMR. Gel formation from several samples, each originating from a curdlan fraction of differing molecular weight, has been observed in order to further clarify the nature of observed phenomena by monitoring their dependence on degree of polymerisation. The results from the complementary techniques described here, in addition to those in existing literature, both for curdlan and for other ss-(1,3) glucans, have been used to build up a consistent framework for the interpretation of results. Broadly, this involves the plasticisation and dissolution of dried material on heating, the time-dependent annealing of native (as biosynthesised) structures, and the trapping of imperfectly formed pseudo-equilibrium states on re-cooling, in concert with the creation of microfibrils and network formation. PMID- 11893390 TI - Salt-induced refolding in different domains of partially folded bovine serum albumin. AB - In our earlier communication on urea denaturation of bovine serum albumin (BSA), we showed significant unfolding of domain III along with domain I prior to intermediate formation around 4.6-5.2 M urea based on the binding results of domain specific ligands:chloroform, bilirubin and diazepam for domains I, II and III, respectively. Here, we present our results on the salt-induced refolding of the two partially folded states of BSA obtained at 4.5 M urea and at pH 3.5, respectively. Both these states were characterized by significant unfolding of both domains I and III as indicated by decreased binding of chloroform and diazepam, respectively. Salt-induced stabilization of partially folded states of BSA was accompanied by nearly complete refolding of both domains I and III as the binding isotherms of chloroform and diazepam obtained in presence of approximately 1.0 M KCl were nearly identical to that obtained with native BSA at pH 7.4. From these observations, it can be concluded that the anion binding sites on serum albumin are not only confined to domain III (C-terminal region) but few sites are also present on domain I (or N-terminal region) of the protein. PMID- 11893392 TI - Catalytic and spectroscopic characterisation of a copper-substituted alcohol dehydrogenase from yeast. AB - Yeast alcohol dehydrogenase (Y-ADH) is widely studied for its biotechnological importance and various attempts to improve its catalytic properties have been made. In this paper, a catalytically active metal-substituted Y-ADH was prepared in vitro by substituting one zinc atom with copper. EPR and Raman spectroscopy suggest that copper maintains the same co-ordination geometry as zinc in native Y ADH. The active Cu-ADH shows lower substrate affinity and lower specific activity (SA) than native ADH, but greater than a previously obtained Co-ADH. Furthermore, Cu-ADH maintains its catalytic efficiency in a wider pH range than native enzyme. PMID- 11893391 TI - HIV-1 encoded virus protein U (Vpu) solution structure of the 41-62 hydrophilic region containing the phosphorylated sites Ser52 and Ser56. AB - Degradation of the HIV receptor CD4 by the proteasome, mediated by the HIV-1 protein Vpu, is crucial for the release of fully infectious virions. To promote CD4 degradation Vpu has to be phosphorylated on a motif DSGXXS, which is conserved in several signalling proteins known to be degraded by the proteasome upon phosphorylation. Such phosphorylation is required for the interaction of Vpu with the ubiquitin ligase SCF-beta-TrCP that triggers CD4 degradation by the proteasome. In the present work, we used two peptides of 22 amino acids between residues 41 and 62 of Vpu. Vpu41-62 was predicted to form an alpha-helix-flexible alpha-helix including the phosphorylation motif DS52GNES56 and Vpu_P41-62 was phosphorylated at the two sites Ser52 and Ser56. We analysed the conformational change induced by the phosphorylation of this peptide on the residues Ser52 and Ser56. Homo- and heteronuclear NMR techniques were used to assess the structural influence of phosphorylation. The spectra of the free peptides, Vpu_P41-62 and Vpu41-62, in both H2O (at pH 3.5 and 7.2) and a 1:1 mixture of H2O and trifluoroethanol were completely assigned by a combined application of several two-dimensional proton NMR methods. Analysis of the short- and medium-range NOE connectivities and of the secondary chemical shifts indicated that the peptide segment (42-49) shows a less well-defined helix propensity. The Vpu_P41-62 domain of residues 50-62 forms a loop with the phosphate group pointing away, a short beta-strand and a flexible extended 'tail' of residues 60-62. Residues 50-60 exhibit alpha-proton NMR secondary chemical shift changes from random coil toward more beta-like structure with the combined (temperature, solvent and pH) NMR and molecular calculation experiments. Differences in this molecular region 50-62 suggest that conformational changes of Vpu_P play an important role in Vpu_P induced degradation of CD4 molecules. PMID- 11893393 TI - Interaction between polylysine monolayer and DNA at the air-water interface. AB - The interaction of a polylysine amphiphile, which consists of a poly-L- or -D lysine (1L or 1D) segment and two long alkyl chains at the C-terminus, with polynucleotides was studied with respect to the highly organized structure of polylysine assemblies on water. The results of surface pressure-area isotherm measurement showed that both of 1L and 1D formed stable monolayers on water in a neutral pH region. The secondary structure of polylysine segment for the surface monolayer was examined by means of circular dichroism and Fourier transform infrared spectroscopies. The helical structure was retained even at neutral pH, at which polylysine has been known to form a complete random coiled conformation in bulk solution. Protonated, positively charged and coiled 1L monolayer could interact electrostatically with guest polyanions including DNA in the subphase, and at the same time the conformation of the polylysine segment was converted from a random coil to an alpha-helix. Deprotonated, helical monolayers did not interact with single stranded polyadenylic acid, but with double stranded DNA. Double stranded DNA was found to interact more strongly with right-handed 1L monolayer than left-handed 1D monolayer. An obvious difference in the melting temperatures for these complexes was observed and discussed on the basis of difference in the interaction mode. PMID- 11893394 TI - Identification and characterization of poly-3-hydroxybutyrate granule-associated protein, PGA12 and PGA16 in Zoogloea ramigera I-16-M. AB - Poly-3-hydroxybutyrate (PHB) granules of Zoogloea ramigera I-16-M contained two major PHB granule-associated proteins (PGA12 and PGA16) as revealed by sodium dodecyl sulfate-polyacrylamide gel elecrophoresis. N-terminal amino acid sequences of these proteins were determined. The genes encoding these proteins were cloned and sequenced. The structural genes of PGA12 and PGA16 were 351 and 447 bp long, which encode polypeptides with deduced molecular masses of 12.3 and 16.0 kDa, respectively. PGA12 and PGA16 were expressed in Escherichia coli. PHB granules were isolated from cells of recombinant strains of E. coli JM109, which harbored and expressed the PHB-synthetic genes of Ralstonia eutropha H16 and PGA12 or PGA16. These PHB granules contained PGA12 or PGA16 as a major protein. The presence of pga12 or pga16 did not affect the amount of PHB synthesized in E. coli. PGA12 and PGA16 bound to crystalline and amorphous PHB granules. PMID- 11893396 TI - Food Safety in Europe (FOSIE): risk assessment of chemicals in food and diet: overall introduction. PMID- 11893397 TI - Hazard identification by methods of animal-based toxicology. AB - This paper is one of several prepared under the project "Food Safety In Europe: Risk Assessment of Chemicals in Food and Diet" (FOSIE), a European Commission Concerted Action Programme, organised by the International Life Sciences Institute, Europe (ILSI). The aim of the FOSIE project is to review the current state of the science of risk assessment of chemicals in food and diet, by consideration of the four stages of risk assessment, that is, hazard identification, hazard characterisation, exposure assessment and risk characterisation. The contribution of animal-based methods in toxicology to hazard identification of chemicals in food and diet is discussed. The importance of first applying existing technical and chemical knowledge to the design of safety testing programs for food chemicals is emphasised. There is consideration of the presently available and commonly used toxicity testing approaches and methodologies, including acute and repeated dose toxicity, reproductive and developmental toxicity, neurotoxicity, genotoxicity, carcinogenicity, immunotoxicity and food allergy. They are considered from the perspective of whether they are appropriate for assessing food chemicals and whether they are adequate to detect currently known or anticipated hazards from food. Gaps in knowledge and future research needs are identified; research on these could lead to improvements in the methods of hazard identification for food chemicals. The potential impact of some emerging techniques and toxicological issues on hazard identification for food chemicals, such as new measurement techniques, the use of transgenic animals, assessment of hormone balance and the possibilities for conducting studies in which common human diseases have been modelled, is also considered. PMID- 11893398 TI - Methods of in vitro toxicology. AB - In vitro methods are common and widely used for screening and ranking chemicals, and have also been taken into account sporadically for risk assessment purposes in the case of food additives. However, the range of food-associated compounds amenable to in vitro toxicology is considered much broader, comprising not only natural ingredients, including those from food preparation, but also compounds formed endogenously after exposure, permissible/authorised chemicals including additives, residues, supplements, chemicals from processing and packaging and contaminants. A major promise of in vitro systems is to obtain mechanism-derived information that is considered pivotal for adequate risk assessment. This paper critically reviews the entire process of risk assessment by in vitro toxicology, encompassing ongoing and future developments, with major emphasis on cytotoxicity, cellular responses, toxicokinetics, modelling, metabolism, cancer related endpoints, developmental toxicity, prediction of allergenicity, and finally, development and application of biomarkers. It describes in depth the use of in vitro methods in strategies for characterising and predicting hazards to the human. Major weaknesses and strengths of these assay systems are addressed, together with some key issues concerning major research priorities to improve hazard identification and characterisation of food-associated chemicals. PMID- 11893399 TI - Hazard characterisation of chemicals in food and diet. dose response, mechanisms and extrapolation issues. AB - Hazard characterisation of low molecular weight chemicals in food and diet generally use a no-observed-adverse-effect level (NOAEL) or a benchmark dose as the starting point. For hazards that are considered not to have thresholds for their mode of action, low-dose extrapolation and other modelling approaches may be applied. The default position is that rodents are good models for humans. However, some chemicals cause species-specific toxicity syndromes. Information on quantitative species differences is used to modify the default uncertainty factors applied to extrapolate from experimental animals to humans. A central theme for extrapolation is unravelling the mode of action for the critical effects observed. Food can be considered as an extremely complex and variable chemical mixture. Interactions among low molecular weight chemicals are expected to be rare given that the exposure levels generally are far below their NOAELs. Hazard characterisation of micronutrients must consider that adverse effects may arise from intakes that are too low (deficiency) as well as too high (toxicity). Interactions between different nutrients may complicate such hazard characterisations. The principle of substantial equivalence can be applied to guide the hazard identification and hazard characterisation of macronutrients and whole foods. Macronutrients and whole foods must be evaluated on a case-by-case basis and cannot follow a routine assessment protocol. PMID- 11893400 TI - Mathematical modelling and quantitative methods. AB - The present review reports on the mathematical methods and statistical techniques presently available for hazard characterisation. The state of the art of mathematical modelling and quantitative methods used currently for regulatory decision-making in Europe and additional potential methods for risk assessment of chemicals in food and diet are described. Existing practices of JECFA, FDA, EPA, etc., are examined for their similarities and differences. A framework is established for the development of new and improved quantitative methodologies. Areas for refinement, improvement and increase of efficiency of each method are identified in a gap analysis. Based on this critical evaluation, needs for future research are defined. It is concluded from our work that mathematical modelling of the dose-response relationship would improve the risk assessment process. An adequate characterisation of the dose-response relationship by mathematical modelling clearly requires the use of a sufficient number of dose groups to achieve a range of different response levels. This need not necessarily lead to an increase in the total number of animals in the study if an appropriate design is used. Chemical-specific data relating to the mode or mechanism of action and/or the toxicokinetics of the chemical should be used for dose-response characterisation whenever possible. It is concluded that a single method of hazard characterisation would not be suitable for all kinds of risk assessments, and that a range of different approaches is necessary so that the method used is the most appropriate for the data available and for the risk characterisation issue. Future refinements to dose-response characterisation should incorporate more clearly the extent of uncertainty and variability in the resulting output. PMID- 11893401 TI - Assessment of intake from the diet. AB - Exposure assessment is one of the key parts of the risk assessment process. Only intake of toxicologically significant amounts can lead to adverse health effects even for a relatively toxic substance. In the case of chemicals in foods this is based on three major aspects: (i) how to determine quantitatively the presence of a chemical in individual foods and diets, including its fate during the processes within the food production chain; (ii) how to determine the consumption patterns of the individual foods containing the relevant chemicals; (iii) how to integrate both the likelihood of consumers eating large amounts of the given foods and of the relevant chemical being present in these foods at high levels. The techniques used for the evaluation of these three aspects have been critically reviewed in this paper to determine those areas where the current approaches provide a solid basis for assessments and those areas where improvements are needed or desirable. For those latter areas, options for improvements are being suggested, including, for example, the development of a pan-European food composition database, activities to understand better effects of processing on individual food chemicals, harmonisation of food consumption survey methods with the option of a regular pan-European survey, evaluation of probabilistic models and the development of models to assess exposure to food allergens. In all three areas, the limitations of the approaches currently used lead to uncertainties which can either cause an over- or underestimation of real intakes and thus risks. Given these imprecisions, risk assessors tend to build in additional uncertainty factors to avoid health-relevant underestimates. This is partly done by using screening methods designed to look for "worst case" situations. Such worse case assumptions lead to intake estimates that are higher than reality. These screening methods are used to screen all those chemicals with a safe intake distribution. For chemicals with a potential risk, more information is needed to allow more refined screening or even the most accurate estimation. More information and more refined methods however, require more resources. The ultimate aims are: (1) to obtain appropriate estimations for the presence and quantity of a given chemical in a food and in the diet in general; (2) to assess the consumption patterns for the foods containing these substances, including especially those parts of the population with high consumption and thus potentially high intakes; and (3) to develop and apply tools to predict reliably the likelihood of high end consumption with the presence of high levels of the relevant substances. It has thus been demonstrated that a tiered approach at all three steps can be helpful to optimise the use of the available resources: if relatively crude tools - designed to provide a "worst case" estimate - do not suggest a toxicologically significant exposure (or a relevant deficit of a particular nutrient) it may not be necessary to use more sophisticated tools. These will be needed if initially high intakes are indicated for at least parts of the population. Existing pragmatic approaches are a first crude step to model food chemical intake. It is recommended to extend, refine and validate this approach in the near future. This has to result in a cost-effective exposure assessment system to be used for existing and potential categories of chemicals. This system of knowledge (with information on sensitivities, accuracy, etc.) will guide future data collection. PMID- 11893402 TI - The contribution of epidemiology. AB - Epidemiologic studies directly contribute data on risk (or benefit) in humans as the investigated species, and in the full food intake range normally encountered by humans. This paper starts with introducing the epidemiologic approach, followed by a discussion of perceived differences between toxicological and epidemiologic risk assessment. Areas of contribution of epidemiology to the risk assessment process are identified, and ideas for tailoring epidemiologic studies to the risk assessment procedures are suggested, dealing with data collection, analyses and reporting of both existing and new epidemiologic studies. The dietary habits and subsequent disease occurrence of over three million people are currently under observation worldwide in cohort studies, offering great potential for use in risk assessment. The use of biomarkers and data on genetic susceptibility are discussed. The paper describes a scheme to classify epidemiologic studies for use in risk assessment, and deals with combining evidence from multiple studies. Using a matrix approach, the potential contribution to each of the steps in the risk assessment process is evaluated for categories of food substances. The contribution to risk assessment of specific food substances depends on the quality of the exposure information. Strengths and weaknesses are summarized. It is concluded that epidemiology can contribute significantly to hazard identification, hazard characterisation and exposure assessment. PMID- 11893403 TI - The FEMA GRAS assessment of pyrazine derivatives used as flavor ingredients. Flavor and Extract Manufacturers Association. AB - This is the fifth in a series of safety evaluations performed by the Expert Panel of the Flavor and Extract Manufacturers Association (FEMA). In 1993, the Panel initiated a comprehensive program to re-evaluate the safety of more than 1700 GRAS flavoring substances under conditions of intended use. Elements that are fundamental to the safety evaluation of flavor ingredients include exposure, structural analogy, metabolism, pharmacokinetics and toxicology. Flavor ingredients are evaluated individually taking into account the available scientific information on the group of structurally related substances. Scientific data relevant to the safety evaluation of the use of pyrazine derivatives as flavoring ingredients is evaluated. PMID- 11893404 TI - Negligible changes in piglet serum clinical indicators or organ weights due to dietary single-cell long-chain polyunsaturated oils. AB - Single-cell oils are currently included in human infant formula as sources of the long-chain polyunsaturates (LCP) docosahexaenoic acid (DHA) and arachidonic acid (AA) in many countries, but have not yet been approved for use in the USA. We prepared four bovine-milk-based formulas with AA/DHA=0, 34/17, 68/34 and 170/85 (mg per 100 kcal formula) provided by two commercial single-cell oils. These levels correspond approximately to 0, 1, 2 and 5 times the concentrations used in infant formulas and, due to greater consumption of formula per unit body weight, resulted in daily consumption of approximately 0, 3, 6 and 16 times those anticipated for human infants. All other dietary fat (47% of calories) was provided by a vegetable oil blend used in commercial human infant formulas. Domestic piglets were allowed to nurse with the sow for 24 h after parturition, then removed to individual cages and maintained on one of the four diets. At 30 days of age the piglets were sacrificed, and serum collected and organs weighed. With litters treated as a blocked variable, no significant differences among groups were found by analysis of variance for the following serum assays: alkaline phosphatase, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), creatinine, albumin, glucose, cholesterol, triglycerides, and total protein. No significant differences were found for hematocrit or body weight. No significant differences were found among groups for weights of liver, brain, heart, lung, spleen, kidneys or lung, analyzed as absolute weight and as a fraction of body weight. Hematoxylin/eosin liver sections examined by light microscopy showed no abnormalities as evaluated by an independent pathologist. DHA content in liver and heart and AA content in heart showed significant dose-related accumulation (P<0.05) and confirmed enhanced tissue accretion of DHA and AA from both oils. We conclude that single-cell oils in formula consumed for 1 month in amounts up to 16-fold greater than proposed for human infants in the USA did not result in clinical chemistry or histopathologic indications of toxic effects in neonatal pigs. PMID- 11893405 TI - Safety evaluation of phytosterol esters. Part 7. Assessment of mutagenic activity of phytosterols, phytosterol esters and the cholesterol derivative, 4-cholesten-3 one. AB - Phytosterol esters are phytosterols derived from vegetable oils following esterification to fatty acids. When phytosterols are added to foods, they inhibit the absorption of dietary and endogenous cholesterol and thereby reduce blood cholesterol concentrations. As part of a comprehensive programme of safety assessment, the mutagenic potential of phytosterols and phytosterol esters has been assessed in a bacterial mutation assay and an in vitro chromosome aberration assay. In addition, an in vitro mammalian cell gene mutation assay and two in vivo mutagenicity studies, namely rat bone marrow micronucleus and liver unscheduled DNA synthesis (UDS) assays, were conducted on phytosterol esters only. Phytosterols and phytosterol esters did not show any evidence of mutagenic activity in any of these assays. A breakdown product of cholesterol is 4 cholesten-3-one and thus the amount of 4-cholesten-3-one in the gut may increase following supplementation of foods with phytosterol-esters. 4-cholesten-3-one had been previously reported as mutagenic but, due to various shortcomings, these data could not be used to assess the mutagenic activity of 4-cholesten-3-one. The mutagenic activity of 4-cholesten-3-one and its major faecal by-products, 5beta cholestan-3-one, was assessed in two in vitro assays, a bacterial mutation assay and an in vitro chromosome aberration assay. Neither 4-cholesten-3-one nor 5beta cholestan-3-one showed evidence of mutagenic activity in these assays. PMID- 11893406 TI - Lack of effects of vitamin E on aluminium-induced deficit of synaptic plasticity in rat dentate gyrus in vivo. AB - Aluminium (Al), has the potential to be neurotoxic in humans and animals, and is present in many manufactured foods and medicines and is also added to drinking water for purification purposes. Our previous study demonstrated that chronic Al exposure induced deficits of both long-term potentiation (LTP) and long-term depression (LTD) of excitatory postsynaptic potential (EPSP) and population spike (PS) in rat dentate gyrus (DG) of hippocampus in vivo (Wang et al., 2001). The purpose of the present study was to investigate whether the Al-induced impairment of synaptic plasticity could be reversed by dietary supplementation with vitamin E (Vit E; alpha-tocopherol). Neonatal Wistar rats were exposed to Al from parturition throughout life by drinking 0.3% aluminium chloride (AlCl3) solution or a diet supplemented with Vit E at 500 microg/g/day with 0.3% AlCl3. The input/output (I/O) function, EPSP and PS were measured in DG area of adult rats (80-100 days of age) in response to stimulation applied to the lateral perforant path. The results showed that: (1) chronic Al exposure reduced the amplitudes of both EPSP LTP (control: 130.4+/-3%, n=7; Al-exposed: 110+/-2%, n=9, P<0.001) and PS LTP (control: 241+/-19%, n=7; Al-exposed: 130+/-7%, n=9, P<0.001) significantly. Vit E had no significant effects on the Al-induced deficits of EPSP LTP (Al-exposed: 110+/-2%, n=9; Al-exposed+Vit E: 112+/-2%, n=8, P>0.05) and PS LTP (Al-exposed: 130+/-7%, n=9; Al-exposed+Vit E: 129+/-4%, n=8; P>0.05); (2) the amplitudes of EPSP LTD (control: 84+/-4%, n=7; Al-exposed: 92+/-7%, n=9, P<0.01) and PS LTD (control: 81+/-4%, n=7; Al-exposed: 98+/-5%, n=9, P<0.001) were also decreased by Al treatment. The impaired EPSP LTD (Al-exposed: 92+/-7%, n=9; Al-exposed+Vit E: 93+/-4%, n=8, P>0.05) and PS LTD (Al-exposed: 98+/-5%, n=9; Al-exposed+Vit E: 94+/-6%, n=8, P>0.05) were also not significantly affected by Vit E treatment. It was suggested that dietary supplementation with Vit E did not reverse the impairment of synaptic plasticity induced by Al in DG in vivo. PMID- 11893407 TI - Effect of T-2 toxin on in vivo lipid peroxidation and vitamin E status in mice. AB - The effects of an acute administration of T-2 toxin on vitamin E status and the corresponding degree of lipid peroxidation, as determined by the plasma and organ content of malondialdehyde (MDA), was studied in mice. The effects of T-2 toxin administration on the body weight and weights of liver, spleen and thymus were also assessed. T-2 toxin was administered in doses ranging from 1 to 6.25 mg/kg body weight, depending on the experiment, while the dietary content of vitamin E ranged from near 0 to 5000 IU/kg. There was a significant decrease in vitamin E content of plasma after the administration of the toxin with the concentrations remaining low for periods as long as 48-72 h. MDA content of liver increased significantly after 24-48 h of toxin administration in contrast to the controls. However, MDA levels returned to the control range after 72 h. The concentrations of MDA in liver were inversely related to the vitamin E content of the diet, and were always higher for the toxin-treated animals (significant linear regression between MDA content of liver and the log10 of vitamin E content of the diet). Weights of spleen and thymus decreased after T-2 toxin administration; however, the weight of liver either increased or did not change in the different experiments. In conclusion, T-2 toxin treatment of mice increased lipid peroxidation in the liver as measured by MDA production. This process was maximal after 48 h of T-2 challenge, and decreased thereafter. Plasma alpha-tocopherol levels decreased as soon as 6 h after the toxin challenge, while MDA did not increase until there was a severe depletion of vitamin E. These changes were accompanied by decrease in weight of spleen and thymus. PMID- 11893409 TI - Pathological evaluation, clinical chemistry and plasma cholecystokinin in neonatal and young miniature swine fed soy trypsin inhibitor from 1 to 39 weeks of age. AB - The potential toxicity of dietary soy trypsin inhibitor (TI) was evaluated in neonatal miniature swine. From 1 to 6 weeks of age, two groups of male piglets were artificially reared in an Autosow and automatically fed either TI or control liquid diet. From 6 to 39 weeks of age, these two groups were fed either TI or control chow diet. A third group, sow control (SC), suckled from birth to 6 weeks of age, were also weaned to control chow from 6 to 39 weeks of age. Clinical chemistry and plasma cholecystokinin (CCK) determined at 6, 18, 30 and 39 weeks of age, and serum amylase activity with gross and histopathological analyses of major organs at 6 and 39 weeks of age are reported. TI had no effect on plasma CCK, serum amylase activity, or numerous clinical chemistry values. TI-fed piglets had a larger relative liver weight at 6 weeks of age. Relative pancreas weight decreased with age but was not affected by TI. Gross and histopathological analyses of major organs, except the spleen, were within normal limits. Increased incidence of extramedullary hematopoiesis was noted in the spleen of the TI group at 6 but not at 39 weeks of age. There was no consistent pattern in immunohistochemical foci for secretin, gastrin releasing polypeptide or CCK, and no change in DNA, RNA, mitotic index or nuclear density of pancreatic cells. At 6 weeks of age, TI increased pancreatic protein and amylase activity but not trypsin or chymotrypsin activity. None of the effects suggested that this dose of TI was toxic to either the neonatal or sexually mature miniature male swine. PMID- 11893408 TI - The Autosow raised miniature swine as a model for assessing the effects of dietary soy trypsin inhibitor. AB - Toxicological effects of dietary soy trypsin inhibitor (TI) were assessed in male miniature swine, a model chosen for its similarities to human digestive physiology and anatomy. The TI preparation was extracted from defatted raw soy flour. From 1 through 5 weeks of age, piglets were automatically fed either a TI liquid diet [Autosow TI group (ASTI)] or a control liquid diet [Autosow control group (ASC)]. From 6 to 39 weeks of age, these animals received either swine chow and TI or swine chow and control article. The TI diets were formulated to contain a TI activity of approximately 500 mg TI/100 g dry matter. A sow control (SC) group suckled from birth to 6 weeks of age and then fed as the ASC group with swine chow plus control article from 6 to 39 weeks of age. The SC piglets grew faster than ASC piglets during postnatal weeks 1 and 2; however, the ASC piglets were significantly heavier than the SC piglets (P=0.001) at 6 weeks of age. Compared with the ASC group, TI caused a moderate decrease in feed consumption and a moderate but reversible decrease in growth from 2 to 5 weeks of age, but not thereafter. Some control and TI-fed Autosow-reared piglets had loose stools until 6 weeks of age; the effect was significantly greater in the TI-fed group. Otherwise, all swine were active and had normal appearance and behavior. PMID- 11893410 TI - Protective effect of Platycodi radix on carbon tetrachloride-induced hepatotoxicity. AB - The protective effects of a Platycodi radix (Changkil: CK), the root of Platycodon grandiflorum A. DC (Campanulaceae) on carbon tetrachloride (CC14) induced hepatotoxicity and the possible mechanisms involved in this protection were investigated in mice. Pretreatment with CK prior to the administration of CC14 significantly prevented the increased serum enzymatic activities of alanine and aspartate aminotransferase in a dose-dependent manner. In addition, pretreatment with CK also significantly prevented the elevation of hepatic malondialdehyde formation and the depletion of reduced glutathione content in the liver of CC14-intoxicated mice. However, hepatic reduced glutathione levels and glutathione S-transferase activities were not affected by treatment with CK alone. CC14-induced hepatotoxicity was also essentially prevented, as indicated by a liver histopathologic study. The effects of CK on the cytochrome P450 (P450) 2E1, the major isozyme involved in CC14 bioactivation were also investigated. Treatment of mice with CK resulted in a significant decrease of P450 2E1 dependent p-nitrophenol and aniline hydroxylation in a dose-dependent manner. CK showed antioxidant effects in FeCl2-ascorbate-induced lipid peroxidation in mice liver homogenate and in superoxide radical scavenging activity. Our results suggest that the protective effects of CK against CC14-induced hepatotoxicity possibly involve mechanisms related to its ability to block P450-mediated CC14 bioactivation and free radical scavenging effects. PMID- 11893411 TI - The in vitro antimutagenic activity of Triphala--an Indian herbal drug. AB - A study to evaluate an antimutagenic potential of water, chloroform and acetone extracts of Triphala has been made in an Ames histidine reversion assay using TA98 and TA100 tester strains of Salmonella typhimurium against the direct-acting mutagens, 4-nitro-o-phenylenediamine (NPD) and sodium azide, and the indirect acting promutagen, 2-aminofluorene (2AF), in the presence of phenobarbitone induced rat hepatic S9. A combination drug 'Triphala' - a composite mixture of Terminalia bellerica, T. chebula and Emblica officinalis, has been used in traditional system of medicine for the treatment of many malaises, such as heart ailments and hepatic diseases. The drug was sequentially extracted with water, acetone and chloroform at room temperature. The study revealed that water extract was ineffective in reducing the revertants induced by the mutagens. The results with chloroform and acetone extracts showed inhibition of mutagenicity induced by both direct and S9-dependent mutagens. A significant inhibition of 98.7% was observed with acetone extract against the revertants induced by S9-dependent mutagen, 2AF, in co-incubation mode of treatment. Various spectroscopic techniques, namely 1H-NMR, normal 13C-NMR, distortionless enhancement by polarization transfer (DEPT-90 and DEPT-135), UV and IR, are under way to identify the polyphenolic compounds from an acetone extract. PMID- 11893412 TI - Toxicity of Australian essential oil Backhousia citriodora (Lemon myrtle). Part 1. Antimicrobial activity and in vitro cytotoxicity. AB - The antimicrobial and toxicological properties of the Australian essential oil, lemon myrtle, (Backhousia citriodora) were investigated. Lemon myrtle oil was shown to possess significant antimicrobial activity against the organisms Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, methicillin-resistant S. aureus (MRSA), Aspergillus niger, Klebsiella pneumoniae and Propionibacterium acnes comparable to its major component-citral. An in vitro toxicological study based on the MTS (3-(4,5-dimethylthiazol-2-yl)-5 (3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) cytotoxicity assay was performed. In vitro cytotoxicity testing indicated that both lemon myrtle oil and citral had a very toxic effect against human cell lines: HepG2 (a hepatocarcinoma derived cell line); F1-73 (a fibroblast cell line derived from normal skin) and primary cell cultures of human skin fibroblasts. Cytotoxicity IC50 (50% inhibitory concentration) values ranged from 0.008 to 0.014% (w/v) at 4 h to 0.003-0.012% (w/v) at 24 h of exposure. The no-observed-adverse-effect level (NOAEL) for lemon myrtle oil was calculated as 0.5 mg/l at 24 h exposure and the RfD (reference dose) was determined as 0.01 mg/l. A product containing 1% lemon myrtle oil was found to be low in toxicity and could potentially be used in the formulation of topical antimicrobial products. PMID- 11893413 TI - Attenuation of bacterial lipopolysaccharide-induced hepatotoxicity by betaine or taurine in rats. AB - The effects of betaine or taurine on hepatotoxicity induced by lipopolysaccharide (LPS) were examined in adult male SD rats. Rats were provided with drinking water containing either 1% betaine or taurine for 2 weeks prior to challenge with LPS (5 mg/kg, iv). Supplementation with betaine or taurine protected the animals from induction of LPS hepatotoxicity as measured by changes in aspartate aminotransferase (AST), alanine aminotransferase (ALT) activities and total bilirubin levels in serum, and hepatic glutathione contents. LPS challenge increased serum TNF-alpha and nitrate/nitrite in rats, which were reduced by betaine or taurine intake. Taurine depletion induced by supply of drinking water containing 3% beta-alanine for 7 days did not enhance the LPS-induced hepatic damage or the decrease in hepatic glutathione level. The results indicate that intake of betaine or taurine attenuates the LPS-induced hepatotoxicity resulting from activation of Kupffer cells. PMID- 11893414 TI - Hypocalcaemia in parental and F1 generation rats treated with sodium fluoride. AB - The potential of sodium fluoride (NaF) to affect serum cations was assessed in the parent (P) and F1 generation rats. The sperm-positive pregnant experimental female rats received 40 mg NaF/kg body weight from day 6 of gestation either up to 21 days of lactation or only up to gestation followed by withdrawal of the treatment during lactation. On day 21 of lactation, blood samples were collected from P and F1 generation rats, allowed to clot and centrifuged at 1000 g for 10 min to obtain serum for analysis of various cations. Statistically significant increases in the concentrations of sodium and potassium in the serum of P and F1 generation rats were observed in the NaF-treated group; however, calcium and phosphorus concentrations were significantly lower than their vehicle control. Withdrawal of NaF treatment during lactation caused significant recovery in sodium, potassium and phosphorus concentrations in P and F1 generation rats as compared with NaF-treated animals. Although statistically significant recovery was not observed, the calcium concentration in P and F1 generation rats was comparatively higher on withdrawal of NaF treatment than in the NaF-treated group. It is concluded that the exposure of 40 mg NaF/kg body weight in pregnant female rats caused significant alterations in cationic concentration which recovered significantly (except calcium) on withdrawal of the treatment. PMID- 11893415 TI - IgG and IgE antibody responses following exposure of Brown Norway rats to trimellitic anhydride: comparison of inhalation and topical exposure. AB - A variety of chemicals can cause sensitisation of the respiratory tract and occupational asthma, including certain acid anhydrides, diisocyanates and reactive dyes. As yet, no well-validated methods are available for the toxicological evaluation of the respiratory sensitising potential of chemicals. One approach which has been explored recently is the evaluation of induced IgE responses or cytokine expression patterns in rats or mice following topical exposure to chemical. Thus, it has been demonstrated that topical exposure of rodents to respiratory sensitising chemicals, but not to contact allergens, causes a dose-dependent and time-related increase in the concentration of total IgE. Using the reference respiratory allergen trimellitic anhydride (TMA), we have considered here the influence of route of exposure on the nature of induced immune responses. Specific IgG and IgE antibody responses and changes in total serum concentration of IgE have been measured following exposure of Brown Norway (BN) starin rats to TMA by topical administration or by inhalation. Exposure to TMA by both routes resulted in the stimulation of specific IgG and IgE antibody, although responses were considerably more vigorous after dermal exposure. Topical treatment also provoked marked and sustained increases in total serum IgE levels, whereas exposure via the respiratory tract stimulated a more transient elevation of this immunoglobulin in a minority of animals which reached statistical significance only at the highest dose group. The lesser vigour of the immune response following inhalation exposure is likely to be related to the considerably lower total antigenic dose which is delivered by this route. Nevertheless, these results show that the nature of immune response with respect to antibody isotype profile provoked by topical administration of TMA is qualitatively comparable with that stimulated by inhalation exposure to the same chemical. For the purposes of hazard assessment and identification of potential chemical respiratory allergens as a function of induced changes in serum IgE concentration, however, the evidence is that topical administration of test material is the preferred route of exposure. PMID- 11893416 TI - Vomitoxin (deoxynivalenol)-mediated inhibition of nuclear protein binding to NRE A, an IL-2 promoter negative regulatory element, in EL-4 cells. AB - Interleukin-2 (IL-2) gene expression is superinduced by the trichothecene mycotoxin vomitoxin (VT, deoxynivalenol) in primary and cloned murine T cells-an activity that relates to this toxin's capacity to inhibit protein synthesis. Binding of the transcription factor ZEB (NIL-2-a) to the negative regulatory element NRE-A plays a critical role in silencing IL-2 expression in the EL-4 cell line, a murine Th1-like lymphoma. The objective of this study was to test the hypothesis that VT impairs NRE-A-binding activity in the EL-4 T cells model. Electrophoretic mobility shift assay (EMSA) in control EL-4 cells revealed a slower migrating band, previously identified as ZEB, and a faster migrating band. VT inhibited NRE-A binding activity in unstimulated EL-4 cells as evidenced by the concentration-dependent reduction of both bands with as little as 50 ng/ml of the toxin being inhibitory. Specificity of NRE-A binding for the two bands was verified by demonstrating (1) competition with excess unlabeled NRE-A probe and absence of competition with mutant NRE-A or unrelated (NF-kappaB) probes and (2) EMSA supershift using antibody specific for ZEB. NRE-A binding activity was also reduced when cells were treated with phorbol 12-myristate 13-acetate (PMA) plus ionomycin (ION). Cotreatment of VT potentiated PMA+ION-mediated reduction of the NRE-A binding activity in concentration-dependent fashion. VT-mediated reduction of NRE-A binding was observed in PMA+ION-stimulated cells as early as 2 h and was still detectable after 24 h. Competitive RT-PCR analysis of mRNA indicated that VT did not reduce ZEB transcript levels. Western analysis of nuclear extracts revealed that, although VT exposure decreased ZEB protein expression, the effects were minimal, required high VT concentrations (500-1000 ng/ml) and were not qualitatively consistent with large concomitant decreases in NRE-A binding activity. Furthermore, VT at levels up to 1000 ng/ml had no effect on ZEB expression in whole cell lysates, suggesting that VT did not selectively inhibit translation of this negative transcription factor. Taken together, these data indicate that the capacity of VT to up-regulate IL-2 expression may relate, in part, to its capacity to decrease ZEB binding to the negative regulatory element within this cytokine's promoter. The inability to associate decreased NRE-A binding with impaired ZEB transcription or translation suggests that post translational modification of this negative transcription factor may be requisite for VT's down-regulatory effects. PMID- 11893417 TI - Organophosphorus pesticides markedly inhibit the activities of natural killer, cytotoxic T lymphocyte and lymphokine-activated killer: a proposed inhibiting mechanism via granzyme inhibition. AB - We have previously found that diisopropyl methylphosphonate, an organophosphorus by-product generated during sarin synthesis in the Tokyo sarin disaster, significantly inhibited natural killer (NK) and cytotoxic T lymphocyte (CTL) activities. In the present study, to investigate whether organophosphorus pesticides (OPs) also affect NK and CTL activities, we firstly examined the effect of five OPs on human NK activity, and then the effect of Dimethyl 2,2 dichlorovinyl phosphate (DDVP), an OP on murine splenic NK, CTL and lymphokine activated killer (LAK), and human LAK activities in vitro. To explore the underlying mechanism of decreased NK activity, we also investigated the effect of 4-(2-aminoethyl) benzenesulfonyl fluoride-HCl (p-ABSF), an inhibitor of serine proteases on NK, LAK and CTL activities, and the effect of DDVP on the activity of granzymes (serine proteases). We found that OPs significantly decreased human NK activity in a dose-dependent manner, but the degree of decrease in NK activity differed among the OPs investigated, and that DDVP significantly decreased NK, LAK and CTL activities in a dose-dependent manner, but the degree of decrease in these activities differed. p-ABSF showed a similar inhibitory pattern to DDVP, and had an additive inhibitory effect with DDVP on NK, LAK and CTL activities. We also found that DDVP significantly inhibited granzyme activity in a dose dependent manner. These findings indicate that OPs significantly decrease NK, LAK and CTL activities in vitro via granzyme inhibition. PMID- 11893418 TI - Development of anti-CD4 MAb hu5A8 for treatment of HIV-1 infection: preclinical assessment in non-human primates. AB - The anti-CD4 MAb 5A8 is a potent inhibitor of CD4-mediated infection of HIV-1. CD4 is obligatory for infection with primary HIV-1 isolates. Humanized 5A8 (hu5A8) was constructed to reduce the potential immunogenicity and enhance the in vivo half-life when used in humans. hu5A8 is a molecularly engineered human IgG4 antibody retaining the binding and functional properties of the murine version of 5A8 (mu5A8). This humanized MAb has been shown to be very effective in inhibiting HIV-1 infection of human CD4+ T cells and macrophages in vitro and to reduce viral load in rhesus monkeys chronically infected with simian immunodeficiency virus (SIV). 5A8 was evaluated in a good laboratory practice (GLP)-compliant tissue cross-reactivity study on human (three donors/37 tissues) and rhesus monkey (two donors/37 tissues) tissues. hu5A8 bound to the surface of human T cells and macrophages, but only to T cells from rhesus monkeys. The antibody did not cross react with other tissues. The highly identical staining patterns of hu5A8 in human and rhesus monkey tissues support the use of rhesus monkeys as a preclinical model for humans. In a GLP-compliant safety study in rhesus monkeys, weekly administration of hu5A8 at 5 mg/kg or 25 mg/kg for 8 weeks was shown to be safe and well tolerated in all monkeys. Although hu5A8 induced anti-hu5A8 antibody response in healthy rhesus monkeys, it was not immunogenic in chimpanzees. Together, the results from these preclinical studies support the studies of the anti-HIV-1 effect of hu5A8 in HIV-1 infected individuals. PMID- 11893419 TI - Genistein and methoxychlor modulate the activity of natural killer cells and the expression of phenotypic markers by thymocytes and splenocytes in F0 and F1 generations of Sprague-Dawley rats. AB - The isoflavone genistein (GE) and methoxychlor (MXC) have been shown to be estrogenic in both in vitro and in vivo experimental systems. The objective of the present study was to evaluate the effects of GE and MXC on the immune system in adult and developing rats and the potential interaction between these compounds in their immunomodulatory actions. Timely pregnant Sprague-Dawley rats were exposed to GE (300 or 800 ppm), MXC (800 ppm), or their combinations in feed starting on day 1 of gestation. The offspring were exposed to these chemicals gestationally and lactationally. Immunological evaluation was performed on postnatal day 22. In F0 females, exposure to GE had no effect on the percentages of thymocyte subsets, but caused a significant decrease in the absolute thymus weight at the 800-ppm dose level. In the spleen, GE did not affect the activity of natural killer cells but induced changes in the percentages of splenic T lymphocyte subsets. Exposure to MXC produced no effect on the immune parameters examined except for a decrease in the percentage of CD4+CD8- thymocytes. Additionally, minimal interaction between GE and MXC was observed. In F(1) males, both GE and MXC decreased the percentage of CD4+CD8- thymocytes, but only GE increased spleen natural killer cell activity. MXC in combination with 300 ppm GE, but not separately, produced significant decreases in the absolute weights of thymus and spleen. In F1 females, GE decreased the percentage of CD4+CD8- thymocytes, increased the percentage of CD4+CD8+ thymocytes, and decreased the activity of spleen natural killer cells. In contrast, MXC increased the percentages of spleen natural killer cells and CD8+ T cells. Overall, the results demonstrate that both GE and MXC can modulate the immune system with greater effects observed in developing rats. Moreover, male and female rats have differential responses to these compounds. A lack of interaction between these two estrogenic chemicals in modulating these immune parameters indicates that their effects on the immune system might involve other mechanisms in addition to the estrogen receptors. PMID- 11893421 TI - Synthesis, characterisation, thermal and explosive properties of 4,6 dinitrobenzofuroxan salts. AB - Two new initiatory molecules, e.g. rubidium and cesium salts of 4,6 dinitrobenzofuroxan (DNBF) have been prepared by reacting sodium salt of 4,6 dinitrobenzofuroxan (DNBF) with rubidium nitrate and cesium nitrate, respectively, at 60 degrees C in aqueous medium. The characterisation of compounds by IR, (1)H-NMR, elemental analysis and metal content is described along with some of the evaluated thermal and explosive properties. The results indicate that cesium salt of DNBF (Cs-DNBF) appears promising initiatory and may suitably replace potassium salt of DNBF (K-DNBF), being used currently in initiatory compositions. PMID- 11893420 TI - BALB/c mice orally pretreated with diclofenac have augmented and accelerated PLNA responses to diclofenac. AB - The nonsteroidal anti-inflammatory drug (NSAID) diclofenac (DF) is associated with idiosyncratic hepatotoxicity and several other distinct hypersensitivity reactions. The mechanism(s) are unknown but evidence suggests both cell-mediated and antibody-mediated immune effector systems may be involved. In the present studies, the immunostimulating potential of DF was evaluated using the direct and TNP-Ficoll (trinitrophenyl (TNP)-Ficoll) popliteal lymph node assays (PLNA). These assays were conducted in naive mice, T-cell-deficient mice, or in mice that had been pretreated with a single oral dose of DF. In naive mice, DF induced a dose-, and time-dependent reaction in the direct PLNA. A significant increase in popliteal lymph node (PLN) weight and PLN cellularity was detected 7 days after the injection of 0.50 and 0.75 mg DF, whereas 0.25 mg DF produced no observable effect. With 0.75 mg, there was a rapid accumulation of cells in the PLN between days 5 and 6, with maximum PLN cellularity observed between days 7 and 10. The immunostimulating effects of DF were significantly attenuated in T-cell-deficient mice. In the TNP-Ficoll PLNA conducted in naive mice, DF caused a dose-dependent increase in PLN cellularity on day 7 with a time-dependent increase in anti-TNP antibody forming cells (AFCs) in the PLN; the reaction was dominated by IgM anti TNP AFCs from day 4 through day 7, but IgG1 anti-TNP AFCs and IgG3 anti-TNP AFCs were detected beginning on day 5 and day 6, respectively. Relative to mice pretreated with vehicle (ddH2O), mice orally pretreated with DF had a significantly greater increase in PLN weight 5 days following the injection of 0.25 mg DF and a significantly greater increase in PLN weight and cellularity 4 days following the injection of 0.50 mg DF. Oral pretreatment with DF had no observable effect on the direct PLN reaction induced following the footpad injection of the irrelevant drugs, D-penicillamine (D-PEN) or streptozotocin. When 0.50 mg DF was co-injected with TNP-Ficoll, mice orally pretreated with DF, compared to vehicle-pretreated mice, and had a significantly greater increase in IgM anti-TNP AFCs on day 4, and a significant increase in both IgG1 and IgG3 anti TNP AFCs on day 7. Additionally, IgG1 anti-TNP AFCs were detected in the PLN of DF-pretreated mice as early as day 4. No differences in anti-TNP AFCs were detected when orally pretreated mice were injected with 0.50 mg D-PEN. Collectively, these results demonstrated that DF (i) is an immunostimulating drug that induced a dose-, time- and T-cell-dependent PLN reaction in naive mice, (ii) provided non-cognate help that produced antibody against co-injected TNP-Ficoll, and (iii) mice orally pretreated with DF had DF-specific increased responsiveness in the direct PLNA, which (iv) resulted in accelerated and augmented AFC production against co-injected TNP-Ficoll. These novel findings suggest that oral administration of DF may result in primed T cells that respond with footpad injection. Thus, the oral pretreatment modification of the PLNA should be further explored as a possible alternative to hypersensitivity testing with drugs administered via the oral route. Additional studies with other compounds known to produce hypersensitivity reactions are needed. PMID- 11893423 TI - Destruction of carbon disulfide in aqueous solutions by sonochemical oxidation. AB - Carbon disulfide (CS(2)) is toxic to animals and aquatic organisms, and can also decompose to carbonyl sulfide (OCS) and hydrogen sulfide (H(2)S) in aqueous environment. The kinetics of the sonochemical degradation of aqueous CS(2) was studied in a batch reactor at 20kHz and 20 degrees C, and the effects of process parameters (e.g. concentration, ultrasonic intensity, irradiating gas) investigated. The concentrations of unbuffered CS(2) solutions used were (6.4 7.0) x 10(-4), 10.5 x 10(-4) and (13.2-13.6) x 10(-4)M and the intensities were varied from 14 to 50W. The reaction rate was found to be zero-order and the rate constant for the degradation at 20 degrees C and 14W in air was 21.1 microM/min using the largest initial concentration range studied. At the same initial concentration range but at 50W (39.47W/m(2)) the degradation rate of CS(2) was 46.7 microM/min, more than two times that at 14W (11.04W/m(2)). The rate of CS(2) sonochemical degradation in the presence of the different gases was in the order of He>air> or =N(2)O>Ar; the rate with helium was found to be about three times that of argon. The formation of sulfate (SO(4)(2-)) as reaction product with air as the irradiating gas was enhanced in the presence of hydrogen peroxide (H(2)O(2)) and inhibited in the presence of 1-butanol. The sonochemical oxidation of CS(2) may prove to be an efficient and environmentally benign way for the removal of this hazardous pollutant from natural water and wastewater. PMID- 11893422 TI - Adsorption of p-chlorophenol from aqueous solutions on bentonite and perlite. AB - The adsorption of p-chlorophenol (p-CP) from aqueous solutions on bentonite and perlite was studied. These materials are available in large quantities in Bulgaria. Model solutions of various concentrations (1-50 mgdm(-3)) were shaken with certain amounts of adsorbent to determine the adsorption capacity of p-CP on bentonite and perlite as well. The influence of several individual variables (initial adsorbate concentration, adsorbent mass) on the rate of uptake of the studied compound on the adsorbent was determined by carrying out experiments at different contact times using the batch adsorber vessel designed according to the standard tank configuration. Rapid adsorption was observed 20-30 min after the beginning for every experiment. After that, the concentration of p-CP in the liquid phase remained constant. The adsorption equilibrium of p-CP on bentonite and perlite was described by the Langmuir and the Freundlich models. A higher adsorption capacity was observed for bentonite (10.63 mgg(-1)) compared to that for perlite (5.84 mgg(-1)). PMID- 11893424 TI - Gas pollutants removal in a single- and two-stage ejector-venturi scrubber. AB - The absorption of SO(2) and NH(3) from the flue gas into NaOH and H(2)SO(4) solutions, respectively has been studied using an industrial scale ejector venturi scrubber. A statistical methodology is presented to characterise the performance of the scrubber by varying several factors such as gas pollutant concentration, air flowrate and absorbing solution flowrate. Some types of venturi tube constructions were assessed, including the use of a two-stage venturi tube. The results showed a strong influence of the liquid scrubbing flowrate on pollutant removal efficiency. The initial pollutant concentration and the gas flowrate had a slight influence. The use of a two-stage venturi tube considerably improved the absorption efficiency, although it increased energy consumption. The results of this study will be applicable to the optimal design of venturi-based absorbers for gaseous pollution control or chemical reactors. PMID- 11893425 TI - Effect of dissolved oxygen concentration on nitrate removal from groundwater using a denitrifying submerged filter. AB - A unidirectional submerged filter system was employed to purify groundwater contaminated with nitrate by biological denitrification. The influence of the concentration of dissolved oxygen (DO) in the process was tested using ethanol, methanol and sucrose as carbon sources. Inorganic-nitrogen removal, growth of the biofilm, platable denitrifying bacteria and nitrate reducing bacteria in biofilm were studied. With regard to the type of electron donor used, the presence of oxygen decreased the removal efficiency of inorganic nitrogen and caused an increase of nitrite concentration in the treated water. These negative effects depended on utilised carbon source. Biological denitrification with alcohols such as ethanol and methanol was less affected by DO than with sucrose. The development of the biofilm was also influenced by the DO concentration as excess O(2) caused reduced biofilm growth. These biofilms developed in oxygen presence had a smaller bacterial density and a lower denitrifying bacteria versus nitrate reducing bacteria ratio, which led to an unfavorable inorganic nitrogen removal and presence of nitrite in the treated water. All these effects are more pronounced when sucrose is used as carbon source. PMID- 11893426 TI - Evaluation of microbial stability of simulated solid and liquid waste forms using a refined biofilm formation method. AB - A refined biofilm formation method was used to evaluate the stability of a simulated liquid waste form containing a simulated liquid waste (salts) and cement in three different proportions, and a simulated solid waste form containing a simulated solid waste (resin) and cement in three different proportions. The experimental samples of all the simulated liquid waste forms showed evidence of microbial growth on them after 3 days of evaluation as indicated by substantial increase in sulfate production, and exhibited considerable instability to microbial degradation as indicated by substantial leaching of calcium. The experimental samples of all the simulated solid waste forms showed evidence of inhibition of growth of Thiobacillus thiooxidans for about 18 days, after which the growth of the microbe became evident in two out of three. Within the growth inhibition period, the differences between experimental and control samples were minor. After the growth of T. thiooxidans became evident, comparatively higher degradations were observed for the experimental samples of the resin containing solid waste forms. PMID- 11893427 TI - Development of a novel electro-dialysis based technique for lead removal from silty clay polluted soil. AB - In this study, a novel electro-dialysis based technique has been developed and used to treat a silty clay soil polluted by lead. The effect of chemical reagents, i.e. tap water at pH 4 (reagent 1) and sodium acetate at pH 5 (reagent 2), on enhancing electro-dialysis extraction of lead from the tested soil was examined. Specimens were prepared by mixing soil with 1000 ppm of lead and were compacted in the dialysate at predetermined dry density and moisture content. Then, specimens were subjected to a predetermined level of current. In the dialysate compartment, anions and cations were removed by charge transport in opposite directions to the anodic and cathodic cells. Meanwhile, in the anodic and cathodic cells, ion concentrations were increased. Thus, concentrated electrolyte streams were produced in alternating cells and cleaned soils were obtained in the dialysate. Both soil pH and lead concentrations were uniformly distributed within the compacted soil specimen during testing. Total lead removal of 80 and 92% was obtained for reagents 1 and 2, respectively. The high removal efficiency was attributed to the separation of electrode reactions from the soil and inclusion of ion selective membranes (ISM), which restrict the movement of counter charged species. PMID- 11893428 TI - Brewery wastewater treatment in a fluidised bed bioreactor. AB - A hydrodynamic characteristic performance of a three phase fluidised bed bioreactor has been studied with brewery wastewater. The influence of operating parameters, such as phase hold up, phase mixing, aspect ratio and superficial gas velocity, on an aerobic biodegradation in a bioreactor of 0.16 m i.d. and 2.7m in height, was analysed. A low-density (960 kg/m(3)) support particle with an internal interstice was employed. The particle and liquid loading were varied in order to determine the effect of phase hold up on bed homogeneity. The ranges in which particle loading and bed height affect fluidisation, and consequently chemical oxygen demand (COD) reduction, were determined. The distributor used in this work was designed such that fluid flow pattern similar to that of a draft tube was induced in the reactor. The low-density particles enabled cost effective operation at a relatively low gas superficial velocity (2.5 cm/s). Aspect ratio significantly influenced the overall bed homogeneity, and the optimum aspect ratio was 10, with volume of the support particles being 21% of the reactor volume. PMID- 11893430 TI - Monocyte phagocytosis as a reliable parameter for predicting early-onset sepsis in very low birthweight infants. AB - BACKGROUND: Septic complications lead to a high mortality rate in very low birthweight infants (VLBWI). Therefore, prognostic markers for the development of sepsis attach importance to start an efficient therapy as early as possible. AIMS: Functional and phenotypical variables of blood monocytes in the cord and peripheral blood were investigated to evaluate the parameters for predicting early-onset and late-onset sepsis (nosocomial infections). STUDY DESIGN: In a prospective study, 25 VLBWI were investigated. METHODS: In the cord blood taken immediately after birth, the capacity of the monocytes to phagocytose non opsonized E. coli bacteria by flow cytometry and the ex-vivo production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta and IL-6 (enzyme-linked immunoassay (ELISA)) after lipopolysaccharide (LPS) stimulation were measured. At the third day, the HLA-DR expression on the monocytes (flow cytometry) and the LPS-induced cytokine production were measured from the peripheral blood. RESULTS: Five VLBWI already developed early septic complications after 24-72 h, while the other three infants had late-onset sepsis 10-18 days after birth. The prognostic significance for early-onset sepsis was highest for the decreased monocyte phagocytic capacity (sensitivity and specificity: 100%) and for the LPS-induced formation of TNF-alpha and IL-1 beta in cord blood. Moreover, in septic VLBWI, the HLA-DR expression on the monocytes was lowered on day 3 after birth. The prognostic significance for late-onset sepsis was highest for TNF-alpha and IL-1 beta levels in the peripheral blood on the third day after birth. CONCLUSIONS: The determination of phagocytosis in the cord blood seems to be a reliable parameter for predicting early-onset sepsis and offers the possibility for a forward start of an antibiotic therapy. PMID- 11893431 TI - Pulmonary prostacyclin is associated with less severe respiratory distress in preterm infants. AB - BACKGROUND: Prostacyclin (PGI(2)) and thromboxane A(2) (TxA(2)) may take part in lung pathology; high concentrations of PGI(2) may protect newborn rabbits against hyperoxic lung injury, and TxA(2) may participate in the development of bronchopulmonary dysplasia (BPD). AIMS: To examine in small preterm infants, the relationship between pulmonary PGI(2) and TxA(2) and respiratory distress during the early postnatal period. METHODS: The stable metabolite of prostacyclin, 6 keto-prostaglandinF(1 alpha), and that of thromboxane A(2), thromboxane B(2), were quantified by radioimmunoassays in 284 samples of tracheal aspirates from 48 infants (GA: 27.4+/-2.1 week, BW 959+/-334 g) during the first 12 postnatal days. RESULTS: Mean concentration of 6-keto-prostaglandinF(1 alpha) was 414+/-31 pg/ml (mean+/-S.E.M.), and of thromboxane B(2) was 418+/-37 pg/ml. Correlations existed between 6-keto-prostaglandinF(1 alpha) and gestational age, birth weight, and the initial arterial-alveolar oxygenation ratio. Negative correlations existed between 6-keto-prostaglandinF(1 alpha) and both mean inspiratory oxygen and duration of mechanical ventilation. Indomethacin treatment was associated with lower pulmonary 6-keto-prostaglandinF(1 alpha), but not with lower TxB(2). Thromboxane B(2) correlated positively with gestational age, birth weight, and initial arterial-alveolar oxygenation ratio, and inversely with duration of mechanical ventilation. CONCLUSIONS: In preterm infants, higher pulmonary 6-keto prostaglandinF(1 alpha) was associated with less severe respiratory distress and with maturity, whereas thromboxane B(2) was associated more strongly with maturity than with respiratory distress. PMID- 11893432 TI - Immunological detection of 4-hydroxynonenal protein adducts in developing pontine and Purkinje neurons and in karyorrhexis in pontosubicular neuronal necrosis. AB - Four-hydroxynonenal (HNE) has been proposed as an important marker of radical induced lipid peroxidation. The principal objective of this study was to assess the occurrence of lipid peroxidation in normal perinatal brain and brains with one form of pontosubicular neuronal necrosis (PSN). Immunochemical studies using an antibody against HNE-modified protein were performed in controls aged from 20 weeks of gestation to 64 years, and patients with PSN. Immunohistochemical study showed developmental and aging changes of positive staining in Purkinje cells and pontine neurons (27 weeks-7 months, 50 and 64 years). In addition, karyorrhectic cells in pontine nuclei with PSN were positively stained. Immunoblotting revealed that a 75-kDa protein, which is speculated to be mitochondrial complex-1 protein, was the most intensely expressed among multiple immunoreactive proteins. Our results identified the presence of oxidative stress in the perinatal neuron, and this oxidative stress may contribute to some forms of karyorrhectic death. PMID- 11893433 TI - Comparison of single and repeated antenatal corticosteroid therapy to prevent neonatal death and morbidity in the preterm infant. AB - INTRODUCTION: In the case of threatened preterm delivery, repeat administration of antenatal corticosteroids is a common practice in women who have not delivered 7-14 days after the first course of corticosteroids. However, the benefits of this policy as compared to single-course administration have not been proven. AIM: Our purpose was to compare neonatal death and morbidity after repeat antenatal courses of corticosteroids with neonatal death and morbidity after a single course. METHODS: We performed a cohort study with matched controls. Neonates treated with repeat antenatal courses of corticosteroids were matched with neonates treated with a single course. Matching criteria were sex, single or multiple gestation, route of delivery, gestational age at delivery and year of birth. Intrauterine growth-restricted infants were matched separately. We excluded neonates with congenital malformation and neonates with an unknown number of antenatal corticosteriod courses. Outcome measures were the incidences of neonatal death, respiratory distress syndrome, intraventricular haemorrhage and necrotizing enterocolitis. RESULTS: From the neonates treated with two or three courses of antenatal corticosteroids, 56 appropriate grown neonates and 24 intrauterine growth-restricted neonates could be matched. The incidences of neonatal death, respiratory distress syndrome, intraventricular haemorrhage and necrotizing enterocolitis did not show statistically significant differences after single and repeat courses of corticosteroids. Appropriate grown and intrauterine growth-restricted neonates showed the same results. CONCLUSION: From our study, it can be concluded that in preterm neonates, repetition of antenatal corticosteroids seems not to improve neonatal outcome. PMID- 11893434 TI - Massage therapy by mothers and trained professionals enhances weight gain in preterm infants. AB - BACKGROUND: The method of "massage therapy" has consistently shown increased weight gain in preterm infants. The weight gain was apparent during massages administered by professionals. AIMS: To replicate the results of increased weight gain in the course of "massage therapy" in preterm infants, and utilize a new, cost-effective application of this method by comparing maternal to nonmaternal administration of the therapy. STUDY DESIGN: Random cluster design. SUBJECTS: The study comprised 57 healthy, preterm infants assigned to three groups: two treatment groups--one in which the mothers performed the massage, and the other in which a professional female figure unrelated to the infant administered the treatment. Both these groups were compared to a control group. RESULTS: Over the 10-day study period, the two treatment groups gained significantly more weight compared to the control group (291.3 and 311.3 vs. 225.5 g, respectively). Calorie intake/kg did not differ between groups. CONCLUSIONS: Mothers are able to achieve the same effect size as that of trained professionals, allowing cost effective application of the treatment within the neonatal intensive care unit. PMID- 11893435 TI - Antenatal supplementation of antioxidant vitamins to reduce the oxidative stress at delivery--a pilot study. AB - BACKGROUND: Preterm infants are at increased risk of oxidative stress and free radical mediated diseases partly related to deficient antioxidant state. The purpose of this study was to investigate if maternal supplementation of antioxidant vitamins prior to delivery would reduce the oxidative stress in the mothers and their infants. STUDY DESIGN: A pilot case-control study. PATIENTS AND METHODS: Five mothers at risk of preterm delivery between 30 and 36 weeks were given a daily oral dose of betacarotene 20 mg, vitamin E 167.8 mg and vitamin C 1000 mg until delivery. Plasma levels of MDA and vitamins A, E and beta-carotene were measured prior to treatment in mothers and at delivery in both mothers and neonates. Seven mother-infant pairs comparable in gestation and birthweight acted as controls. RESULTS: In the supplemented group, median maternal plasma MDA at delivery was significantly lower compared to the pretreatment level (1.9 vs. 3.2 micromol/l, p=0.04) and it was also lower than the control group (1.9 vs. 3.65 micromol/l, p<0.001). In the supplemented group, median maternal plasma vitamin E at delivery was significantly higher than the levels prior to treatment (46 vs. 31 micromol/l, p=0.007) in the same group and those at delivery in the control group (46 vs. 30 micromol/l, p=0.03). There was a trend of lower plasma MDA and higher vitamin E at birth in infants born to supplemented mothers, but it did not reach statistical significance. CONCLUSION: A short supplementation of multiple antioxidant vitamins to a small sample of preterm pregnant women reduced the oxidative stress at delivery in mothers and probably in their neonates. Larger studies probably using larger doses are needed to evaluate the efficacy of this strategy. PMID- 11893436 TI - Maternal serum urea resistant alkaline phosphatase in Down syndrome pregnancy. AB - BACKGROUND: The normal levels of alkaline phosphatase (AP) activity in maternal serum are virtually the same as those observed in Down syndrome (DS) pregnancies at 14-20 weeks' gestation. Using urea inhibition of AP, we observed an atypical AP isoenzyme in the neutrophils of mothers with trisomy 21 fetuses. AIM: To assess the use of urea as a selective inhibitor of serum AP in order to seek a possible diagnostic difference between normal and DS pregnancies. STUDY DESIGN AND SUBJECTS: Serum AP samples from 24 DS pregnancies and 204 control cases were examined at 12-22 weeks' gestation with and without 2.5 M urea AP inhibition at 18 degrees C for 2 h. The levels of AP activity obtained without urea and the percentage urea AP inhibition were analyzed in the two groups. RESULTS: Without urea treatment, no significant difference of total alkaline phosphatase activity levels was detected between the 204 normal controls and the 24 DS samples. Using 2.5 M urea AP inhibition, after 120 min of exposure at 18 degrees C, the residual activity, as a percentage of initial values of AP, showed significantly higher resistance in the DS samples (> or = 50 IU/l of total AP activity) at 15-22 weeks' gestation. However, at 12-14 weeks (< or = 45 IU/l of total AP activity), no significant difference was found between the DS and control cases. CONCLUSION: Serum urea resistant alkaline phosphatase in DS pregnancies showed a significant difference only at 15-22 weeks' gestation, compared with normal controls. PMID- 11893437 TI - sCD31/sPECAM-1 levels in breast milk and sera of mother-infant pairs in the early postpartum period. AB - Immunomediators seem to have a central role in the immune system of both human milk and newborn infants. CD31/PECAM-1 is an adhesion molecule, member of Ig gene superfamily, mediating cell-cell adhesion in both homophilic and heterophilic ways. Levels of the soluble form of PECAM-1 (sPECAM-1) were evaluated on the 2nd and 5th day postpartum in breast milk and serum paired samples from 20 lactating women as well as in time-matched serum from their single, term, healthy neonates. Concentrations of sPECAM-1 in breast milk (median, range) on both the 2nd (2.05 ng/ml, 0.0-7.2) and 5th day postpartum (0.89 ng/ml, 0.0-3.6) were about 10 and 20 times lower than those (mean +/- SD) in controls (healthy adults) (19.83 +/- 5.17, p<7 x 10(-8)), showing a significant fall from the 2nd to the 5th day postpartum (p<0.0005). Maternal serum sPECAM-1 values (mean +/- SD) were significantly lower on the 2nd day postpartum (14.21 +/- 5.15 ng/ml) than those in controls (p<0.002), but reached control values on the 5th day postpartum after a significant rise (p<0.0075). Neonatal serum sPECAM-1 values with no significant difference between the 2nd (14.4 +/- 4.11 ng/ml) and 5th day of life (14.54 +/- 4.99 ng/ml) were significantly lower than those in controls (p<0.002). Values of sPECAM-1 in milk and sera of lactating mothers and their neonates on the 2nd day postpartum depended on the mode of delivery, being significantly lower after caesarean section (p<0.034, p<0.0075 and p<0.035, respectively). In conclusion, our findings in the early postpartum period demonstrate that: (a) sPECAM-1 is present in human milk in low and decreasing concentrations; (b) the shedding of sPECAM-1 is an established component of the neonatal immune system from birth, though in lower concentrations than in adults, possibly reflecting its immaturity; and (c) the mode of delivery has a significant effect on sPECAM-1 values in milk and sera of lactating mothers and their neonates; the lower values after caesarean section may reveal a deranged endothelial homeostasis. PMID- 11893438 TI - The influence of ciclosporine A on the vasoactive effects of serotonin in in vitro perfused human umbilical arteries. AB - BACKGROUND: Pregnancy is feasible in organ-transplanted women, but little is known about possible effects of ciclosporine A on the circulation in the fetus and placenta. AIM: To investigate the influence of ciclosporine A (CsA) on the vasoactive effects of serotonin in human umbilical arteries. STUDY DESIGN AND SUBJECTS: In vitro perfusion was performed in umbilical cord segments from seven organ-transplanted patients on CsA based immunosuppression and in 17 cords from uncomplicated pregnancies. Serotonin was administered in stepwise increasing concentrations from 10(-10) to 10(-5) M. In preparations from normal pregnancies, serotonin 10(-7) M, was administered before and 30 min after the start of a continuous CsA infusion (1.0 mg/l). The influence of CsA 0.1 or 1.0 mg/l on the basal, unstimulated perfusion pressure was investigated in separate experiments. OUTCOME MEASURES: Changes in perfusion pressure due to constrictory or dilatatory responses. RESULTS: In all preparations from the organ-transplanted patients, serotonin induced a constrictory response that was non-significantly lower than that observed in the control group. The frequency of a dilatatory response preceding the vasoconstriction was 3/7 and 12/17 (non-significant) in the CsA treated and control groups, respectively. In the experiments with CsA administration, a non-significant increase in the constrictory serotonin response was observed as compared to the control experiments. CsA did not alter the basal, unstimulated perfusion pressure. CONCLUSION: CsA did not have any significant influence on the vasoactive effect of serotonin in human umbilical arteries perfused in vitro. PMID- 11893439 TI - Interleukin-16 in tracheal aspirate fluids of newborn infants. AB - BACKGROUND: It is currently unknown if interleukin (IL)-16 exists in the lungs of ventilated infants, and because the predominant cells in the airways of infants with CLD are CD4+ macrophages, we hypothesized that IL-16 plays a role as a pro inflammatory mediator in lung inflammation. AIMS: To examine if IL-16, a chemoattractant for CD4+ cells, is detectable in airway secretions of ventilated newborns. Its presence may be associated with lung inflammatory responses. STUDY DESIGN: Cohort cross-sectional study. SUBJECTS: Thirty-four mechanically ventilated newborn infants. MAIN OUTCOME MEASURES: Tracheal fluid (TF) specimens collected during the first month of life were examined for cell differentials determined from cytospin slides and supernatant was analyzed by ELISA for IL-16. RESULTS: Eighty-three cross-sectional tracheal fluid (TF) specimens were analyzed. Eleven of the 27 preterm but none of the 7 term infants developed chronic lung disease (CLD). IL-16, ranging from 203 to 42,073 pg/ml, was detected in 16 of the 46 specimens obtained from CLD infants, 1 of the 30 specimens from 16 non-CLD preterm and 2 of the 7 specimens from 7 term infants (p<0.001). Leukocyte counts (median 16.6 vs. 2.0 x 10(-9)/l, p<0.0001) and percentage neutrophils (median 93% vs. 73%, p<0.001) were higher in IL-16 positive specimens. CONCLUSION: IL-16 is detectable within the airway secretions of ventilated newborn infants and its presence is associated with a neutrophilic infiltration. Further studies are required to investigate its role in chronic inflammation in CLD. PMID- 11893440 TI - Maternal anxiety in late pregnancy: effect on fetal movements and fetal heart rate. AB - AIM: To determine whether maternal state and trait anxiety levels affect fetal movements or fetal heart rate (FHR) in the third trimester. SUBJECTS: Forty-one healthy pregnant nulliparous women not on medication and with a singleton pregnancy. STUDY DESIGN: Maternal anxiety was assessed using the Spielberger State- Trait Anxiety Inventory (Form Y) at 36 gestational weeks. The fetuses of the women were examined at 37-40 gestational weeks with ultrasound observation of fetal movements and cardiotocography (CTG). The results of the fetal examinations were compared between women with low and high anxiety scores (low scores being defined as scores below the median and high scores as scores equal to or above the median of the study population), and correlation analyses between anxiety scores and the outcome variables were performed. OUTCOME MEASURES: The presence and duration (expressed as a percentage of the total examination time) of FHR patterns A, B, C, and D, the percentage duration of fetal movements in each FHR pattern, baseline FHR and FHR variability in each FHR pattern. RESULTS: The presence of FHR patterns A, B, C, and D, the duration of FHR patterns A, B, and C, FHR variability in FHR patterns A, B, and C, baseline FHR and the percentage duration of fetal movements in each FHR pattern did not differ between women with low and high state and trait anxiety scores. In fetuses with FHR pattern D, the duration of FHR pattern D increased with increasing maternal trait anxiety scores, (rho=0.88; p=0.008), and FHR variability in FHR pattern D increased with maternal state and trait anxiety scores (r=0.86, p=0.01; r=0.96, p=0.001). CONCLUSION: Maternal anxiety does not seem to affect fetal movements or baseline FHR in late pregnancy, but there is a possible association between maternal anxiety and the duration of FHR pattern D and FHR variability in FHR pattern D. PMID- 11893441 TI - Growth and bone mineralization of young adults weighing less than 1500 g at birth. AB - BACKGROUND: Preterm infants are at risk for suboptimal growth and bone mineralization compared to infants born at term but long-term outcomes into early adulthood are unclear. AIMS: To determine (1) if growth and nutrition in the first year of life significantly predict the outcomes measured at adulthood and (2) whole body and regional bone mineral content (BMC) of young adults who were born preterm and weighing <1500 g. STUDY DESIGN AND SUBJECTS: In this descriptive follow-up study, subjects were born preterm and weighing <1500 g (n=25, 17.2+/ 1.2 years of age) and originally participated in a 1-year follow-up study of infant growth or subjects born at term (n=25, 17.3+/-1.4 years of age). OUTCOME MEASURES: In the preterm group, relationships of growth and nutrition in the first year of life with adult anthropometry and BMC were identified using correlation and regression analysis. Birth groups were compared for measurements of anthropometry and whole body and regional BMC obtained at adulthood using t tests. RESULTS: After correcting for the effects of bone area using regression, rate of weight gain had a positive relationship and days to regain birth weight a negative relationship to adult BMC. Young adults, born preterm, were significantly shorter with lower whole body BMC than of those born at term, but BMC was appropriate for size. CONCLUSIONS: Growth early in life predicts subsequent attainment of growth and bone mass. Premature birth results in lower attainment of height achieved by young adult age but bone mass is appropriate for body size. PMID- 11893443 TI - Bilateral cholesteatoma and habitual sniffing. AB - OBJECTIVE: To examine the clinical findings of acquired bilateral cholesteatoma with special reference to incidence of habitual sniffing and sniff-related negative middle ear pressure. METHODS: Eighty-eight fresh cases of unilateral cholesteatoma and 33 fresh cases of bilateral cholesteatoma, which were operated on at Department of Otolaryngology, Hyogo College of Medicine, were examined in this study. Responses to a detailed questionnaire were obtained from the patients concerning about the habit of habitual sniffing to relieve aural symptoms such as aural fullness, autophonia or hyperacusis. The same questionnaire was obtained from unilateral cholesteatoma patients to compare the incidence of habitual sniffing with that of patients with bilateral cholesteatoma. We measured the negative middle ear pressure at the time of sniffing by using TTAG (tubo-tympano aerodynamic graphy, Nagashima Co. Ltd, Tokyo). We also compared the positive percentage of the sniff test in bilateral cholesteatoma with in unilateral cholesteatoma and normal controls. Sniff test was performed in 30 patients with bilateral cholesteatoma, 20 patients with unilateral cholesteatoma and 20 normal controls. RESULTS: In 33 patients with bilateral cholesteatoma (66 ears), 57 ears had the pars flaccida type (86.4%) and 9 ears had the pars tensa type (13.6%). Cholesteatoma of pars flaccida type were predominant in bilateral cholesteatoma. The rate of habitual sniffing of bilateral cholesteatoma (23/33, 69.7%) was significantly higher than that of unilateral cholesteatoma (21/88, 23.9%). The incidence of positive sniff test in bilateral cholesteatoma (19/30, 63.3%) was significantly higher than in unilateral one (6/20, 30%) and normal control (3/20, 15%). CONCLUSIONS: Habitual sniffing was closely related to the pathogenesis of bilateral cholesteatoma, especially in cases with bilateral pars flaccida type. PMID- 11893442 TI - Auditory-evoked cerebral oxygenation changes in hypoxic-ischemic encephalopathy of newborn infants monitored by near infrared spectroscopy. AB - Recent neuronal activation studies have demonstrated the presence of regional cerebral blood flow (rCBF) increases in response to neuronal activation in normal newborns. In the present study, using near infrared spectroscopy (NIRS), we evaluated the evoked cerebral blood oxygenation (CBO) changes in hypoxic-ischemic encephalopathy (HIE) of newborns. We studied 20 normal newborns and 22 HIE newborns; mild HIE (n=9), moderate HIE (n=7), and severe HIE (n=6). The babies were from 1 to 3 days postdelivery. We measured the concentration changes of deoxyhemoglobin (Deoxy-Hb), oxyhemoglobin (Oxy-Hb), and total hemoglobin (Total Hb) induced by auditory stimulation in the frontal lobes. The normal and HIE groups showed different Oxy-Hb and Total-Hb responses. In normal newborns, 19 out of 20 normal subjects showed increases of Oxy-Hb and Total-Hb, whereas 14 out of 22 subjects showed decreases of Oxy-Hb and Total-Hb during the stimulation (chi(2)=19.95, p<0.001). In addition, there was a strong negative correlation between HIE severity and changes of Total-Hb (r=-0.73, p<0.001). These results suggest that infants with HIE have decreased rCBF in the frontal lobes during auditory stimulation. PMID- 11893444 TI - Hearing loss and tinnitus in Meniere's disease. AB - OBJECTIVES: To characterize hearing loss, tinnitus and associative factors in Meniere's disease. METHODS: From our vertigo database consisting of 1356 patients, we retrieved 243 patients with Meniere's disease (MD). RESULTS: Hearing loss was the initial symptom in 13% of cases. Altogether 64% (n=133) of the patients stated that their hearing was reduced during the vertigo attack. The hearing deteriorated more likely during the vertigo attack if the vertigo attack was very intense [r(206)=0.19]. Tinnitus was the initial symptom in 5% of cases. The tinnitus was mild in 38% (n=90), moderate in 32% (n=76) and severe in 30% (n=72) of patients. The intensity of tinnitus correlated with the occurrence of drop attacks [r(237)=0.29], vertigo provoked by head positioning [r(235)=0.25], by physical activity [r(230)=0.33], or by pressure changes [r(239)=0.27]. CONCLUSION: Prolonged disease causes deterioration of hearing. Intense tinnitus is common in MD and is more often seen in late stage of the disease. PMID- 11893445 TI - Effect of background noise on perception of English speech for Japanese listeners. AB - OBJECTIVES: To avoid the accidents induced by confusions of non-native speech perception in the presence of noise at work, this study examined the effect of background noise on the discrimination of English speech for Japanese listeners. METHODS: Normal hearing Japanese subjects, who have English learning experience to the level of college graduates, were tested with the CID W-22 word list in quiet settings and in the presence of white, low-frequency weighted (pink noise), or aircraft noise. RESULTS: The discrimination scores were reduced as S/N ratio decreased, and more adversely affected by background white noise than pink or aircraft noise. The individual variability of English word discrimination scores for Japanese listeners was much larger as the S/N ratio decreased. The typical confusions among phoneme were /m/, /n/, and /s/. CONCLUSIONS: The results of this study suggested that ineffective use of speech cues and difference in composition of English phonemes from Japanese could be responsible for degraded speech discrimination of non-native speech in noise. Moreover, it was expected that Japanese workers with age related hearing loss at higher frequencies might have great difficulty in perceiving English speech sounds in the presence of noise. PMID- 11893446 TI - Experimental otitis media with effusion induced by leukotriene D4. AB - OBJECTIVE: inflammatory mediators such as prostaglandins (PGs), leukotrienes (LTs), and platelet-activating factor (PAF) have been identified in human middle ear effusions (MEEs), as well as in the experimentally induced MEEs of animals. However, the exact function of LTs in the middle ear cavity is still unknown. In this study, the role of LTs was investigated using an experimental model in which OME was induced by LTD4. METHODS: to examine whether leukotrienes (LTs) induce otitis media with effusion (OME), 2x10(-6), 2x10(-5), 1x10(-4) and 2x10(-4) M of LTD4 was injected into the rat ear. The severity of OME was assessed based on the histological findings. The concentrations of IL1-beta, TNF-alpha, and GRO/CINC-1 in MEE were also measured. Additionally the therapeutic efficacy of a specific LTs antagonist, pranlukast on experimental OME was investigated. RESULTS: all ears developed middle ear effusion (MEE) within 24 h and about 50% of the animals continued to demonstrate MEE for 14 days in the 1x10(-4) and 2x10(-4) M groups. The cytokine levels seemed to correspond well with the persistence of OME. The oral administration of a specific LTs antagonist, pranlukast, was found to alleviate the experimental OME. CONCLUSION: these findings suggest that LTs appear to plays an important role in the pathogenesis of OME. PMID- 11893447 TI - Autoclaving the ossicles provides safe autografts in cholesteatoma. AB - OBJECTIVE: The choice of the graft in ossicular chain reconstruction during middle ear surgery for cholesteatoma is a subject still discussed on. In order to clarify the discussion of reuse of the autologous ossicles obtained during middle ear surgery for cholesteatoma, we evaluated the probability of residual disease histologically and the safety of the ossicles after autoclavization, the most promoting alternative method to eradicate residual cholesteatoma and infection on them. METHODS: The specimens used in this study were eroded twenty-seven ossicles (22 incuses, 5 malleoli) which were removed from the 27 consecutive patients operated because of cholesteatomatous middle ear disease. They were grouped as follows: Group 1, Fifteen ossicles examined histopathologically directly. Group 2, Five ossicles autoclaved for 20 min at 134 degrees C and then examined histopathologically. Group 3, Five ossicles autoclaved for 20 min at 134 degrees C after mechanical surface cleaning by a fine diamond drill, examined histopathologically. Group 4, Two ossicles removed from two different patients were placed in their mastoid cavities in order to be examined after access in the second-look operation. While one ossicle was only autoclaved, the other was mechanically cleaned by a drill before autoclavization (for 20 min at 134 degrees C). The ossicles were examined histopathologically after the removal at the second stage operation performed 12 months later. RESULTS: In Group 1, all ossicles showed evidence of periosteal thickening. Additional findings were surface cholesteatoma or epithelia in 13 ossicles, surface inflammation in 12 ossicles, granulation tissue in 10 ossicles, osteitis in six ossicles. In Group 2, all five ossicles had preserved their lamellar structure but, no vital cells were seen. The lacunes that had the osteocytes was almost completely empty. The inflammatory cells were eliminated from the ossicles. In Group 3, ossicles were found well preserved with their lamellar structures and contours, with empty lacunes and eliminated inflammatory cells. In Group 4, in two ossicles of this group the lacunes were replaced by the new migrated viable osteocytes with evidence of new bone formation and neovascularisation. No new inflammatory focus or epithelia were found on the surfaces of the ossicles. The shape and the contour of the ossicles remained unchanged. CONCLUSION: In cholesteatoma surgery, ossicles with minimal erosion and adequate thickness can be used after autoclavization. In this study, it was observed histopathologically that the autoclaving autologous ossicles before ossiculoplasty in cholesteatomatous middle ear is a safe and reliable method. PMID- 11893448 TI - Vertical eye-movement oscillation with a frequency double that of lateral linear acceleration in patients with long-standing unilateral vestibular loss. AB - OBJECTIVE: To examine our hypothesis that an unusual floating or tilting sensation claimed by patients with long-standing unilateral vestibular loss might be attributed to incomplete central compensation in the otolith system. METHODS: Seven patients who were with or without symptoms for 6-101 months after intratympanic gentamicin therapy for unilateral endolymphatic hydrops were sinusoidally exposed to lateral linear acceleration, and their compensatory eye movements were compared with those of 18 normal controls, using electro oculography (EOG). The subjects, secured firmly in the chair of a linear accelerator (sled), were oscillated at three different G-loadings of 0.1 (0.11 Hz), 0.2 (0.16 Hz) and 0.3 G (0.19 Hz), respectively. During displacement, they gazed at a real (visible) or imaginary target which was located on the wall (earth-fixed), or moved on the wall in sync with the sled (body-fixed). RESULTS: Vertical EOG (V-EOG) with a frequency two-fold that of the stimulus frequency was characteristically evoked in all patients, but in none of the normal controls. One of the doubled V-EOG was larger in amplitude than the other, when the stimulus acceleration was directed from the affected ear to the intact ear in those patients whose symptoms still remained. Such a directional difference tended to be greater with the imaginary target than with the visible target. In the horizontal EOG, there was no marked difference between the patients and normal controls. Nobody reported tilt perception but only horizontal translation during the sled displacement, except one patient who was examined twice; a roll tilt was perceived in the first examination when she still had symptoms, but no tilt sensation in the second one when she no longer had symptoms. CONCLUSION: It was suggested that incomplete compensation in the otolith system could cause unusual sensations even after long-standing unilateral vestibular loss, and that V-EOG induced by a sled might be helpful in evaluating functional recovery. PMID- 11893449 TI - The role of free oxygen radicals in noise induced hearing loss: effects of melatonin and methylprednisolone. AB - The aim of this study was to investigate the role of cochlear damage caused by free oxygen radicals occurring as a result of exposure to noise and to determine the prophylactic effects of melatonin and methylprednisolone. Fifty male albino guinea pigs were randomly divided into five groups. All groups were exposed to 60 h of continuous wide band noise at 100+/-2 dB, except group I. Group I was not exposed to noise or treated with drugs. Group II was exposed to noise and not treated with drugs. Group III was exposed to noise and treated with melatonin. Group IV was exposed to noise and treated with methylprednisolone. Group V was exposed to noise and treated with melatonin and methylprednisolone. A high dose of 40 mg/kg methylprednisolone and/or 20 mg/kg melatonin were administered intramuscularly 24 h before exposure to noise, immediately before noise exposure and once a day until noise exposure was completed. Just after the noise ended, guinea pigs were decapitated. Venous blood was obtained into tubes with EDTA and it was used to measure activity levels of plasma malondialdehyde, erythrocyte glutathione peroxidase and the cochlear tissue malondialdehyde. After the noise ended, in comparison group II with I; it was found that the malondialdehyde activity of the plasma and tissue had increased, the erythrocyte glutathione peroxidase activity levels had decreased and consequently, hearing thresholds had increased (P<0.01). A significant difference was found in the malondialdehyde and erythrocyte glutathione peroxidase activity levels between groups II and III (P<0.01) and the hearing thresholds exhibited a parallel trend (P<0.05). The hearing threshold and malondialdehyde activity levels obtained from groups IV and V were found to be similar to those of group II (P>0.05). As a conclusion, we suggest that the use of methlyprednisolone in order to prevent the cochlear damage caused by noise does not provide sufficient prophylaxy, however the use of melatonin provides a more effective prophylaxy, thus being a promising alternative. PMID- 11893450 TI - Neoglottic formation from posterior pharyngeal wall conserved in surgery for hypopharyngeal cancer. AB - OBJECTIVE: To describe a new treatment modality of hypopharyngeal cancer consisting of total laryngectomy plus partial pharyngectomy (TLPP) conserving the posterior wall of the pharynx vertically for voice restoration. METHODS: Review of hospital charts, TLPP was undertaken in 15 of 54 patients. Surgical modalities of reconstruction subsequent to TLPP were indicated on the basis of the width of posterior pharyngeal wall conserved during surgery. Posterior pharyngeal walls of width 3 cm or larger were sutured in primary closure. If the width of posterior wall was less than 3 cm, a free forearm flap or free jejunal flap was patched to the wall. Tracheo-esophageal shunt with a voice prosthesis was performed 3 weeks after surgery. RESULTS: The Kaplan-Meier method indicated no difference in survival rate between patients with TLPP (46.4%) and the remaining patients (47.4%). Nine of 15 patients with TLPP (two patients with primary closure, three with free forearm flap, and four with free jejunal flap) were examined for voice restoration and fluoroscopy of the neopharynx. Eight of the nine patients, in whom more than 2 cm of the posterior pharyngeal wall had been conserved, demonstrated a good speech rating, maximum phonation time and neoglottic formation by the posterior pharyngeal wall. CONCLUSION: The combination of conservation of the posterior pharyngeal wall, patch graft and a voice prosthesis is a useful method that offers sufficient quality of phonation without deterioration of survival rate for patients with hypopharyngeal cancer. PMID- 11893451 TI - Excessive post-tonsillectomy hemorrhage requiring ligature of the external carotid artery. AB - Ligature of the external carotid artery (LECA) is the method of choice in patients with excessive post-tonsillectomy hemorrhage. This retrospective study was undertaken to evaluate the incidence, characteristics and warning signs of excessive post-tonsillectomy hemorrhage. BASIC PROCEDURES: Between January 1988 and December 2000, a total of 25 patients had to be treated by LECA. Tonsillectomy had been previously performed in seven patients at our department (group A) and in 18 patients elsewhere (group B). MAIN FINDINGS: LECA was performed in most cases 6 (group A) and 11 days (group B) after tonsillectomy. There was one case with lethal outcome. A total of 12 patients (group B) had been operated by two surgeons. PRINCIPAL CONCLUSIONS: Excessive bleeding following tonsillectomy may occur as delayed bleeding, abrupt and require immediate LECA and blood transfusion. Prior recurrent episodes of bleeding can be a warning sign. Anatomical vascular abnormalities have to be considered. Inpatient policy in these underestimated cases was life-saving. PMID- 11893452 TI - Abnormalities of molecular regulators of proliferation and apoptosis in carcinoma of the oral cavity and oropharynx. AB - OBJECTIVE: Abnormalities in genes regulating cell proliferation and death may affect disease outcome in squamous cell carcinoma (SCC) of the head and neck. METHODS: Proliferative activity (Histone H3 in-situ-hybridization (HISH) labeling index (LI)) and the genes and/or gene products of Cyclin D-1, c-erbB-2, Bcl-2, p21, and p53, were investigated in 35 patients with SCC of the oral cavity and oropharynx, previously studied for p27 expression. RESULTS: Overexpression or very low expression of Cyclin D-1 was associated with unfavorable disease outcome and shorter time-to-recurrence. High c-erbB-2 expression was significantly associated with shorter overall survival and was synergistic with low p27 expression. Bcl-2, HISH LI, p21 expression, and p53 mutation and protein analysis were not significantly predictive, but there were trends suggesting shorter disease-free/overall survival for patients with undetectable Bcl-2, high HISH, and mutant p53. CONCLUSIONS: Several cell proliferation and death regulators appeared to predict disease outcome. Limited evidence of cooperativeness among regulators was also seen. PMID- 11893453 TI - Significance of epidermal growth factor receptor and tumor associated tissue eosinophilia in the prognosis of patients with nasopharyngeal carcinoma. AB - OBJECTIVE: The expression of epidermal growth factor receptor (EGFR) is one of the indicators, which can predict the malignant potential of squamous cell carcinoma of the head and neck (HNSCC). On the other hand, previous histological studies have proved tumor-associated tissue eosinophilia (TATE) to be a favorable prognostic indicator for HNSCC. We studied the prognostic significance of co expression of EGFR and TATE. METHODS: We examined the expression of EGFR and TATE in 53 patients with nasopharyngeal carcinoma (NPC). Biopsy specimens were subjected to immunohistochemical-staining for EGFR expression and Luna-staining for TATE. EGFR staining was considered negative when immuno-stained cells were less than 25% in a field. TATE was divided into four grades as grade 0 for 0-2 eosinophils in a high power field, grade 1 for 3-9, grade 2 for 10-29, and grade 3 for 30 or more. RESULTS: In terms of TATE expression, 27 patients were classified as grade 0, 12 as grade 1, six as grade 2, and eight as grade 3. Forty four patients were EGFR positive and nine were negative. We found no statistical significance in the distribution of EGFR positivity and TATE grades. Among EGFR positive patients, 5 year survival rates were significantly better in TATE positive (grades 1, 2, 3) patients than in TATE-negative (grade 0) patients (P=0.0139). CONCLUSION: Eosinophils may be activated in the tumor tissue, in which the expression of EGFR is up-regulated. This suggests that the activated eosinophils in EGFR-positive tumors resulted in better prognoses. TATE infiltration and EGFR expression may be closely related to the malignant potential of NPC, and co-expression of TATE and EGFR may be an important prognostic factor. PMID- 11893454 TI - Feline immunodeficiency virus-mediated gene therapy of middle ear mucosa cells. AB - HYPOTHESIS: To investigate the feasibility of gene therapy of the middle ear mucosa using a novel vector. BACKGROUND: Given present medications are unable to affect chronic otitis media, cholesteatoma, or tympanic membrane perforation, newer methods of treatment like gene therapy for these diseases must be explored. These genes can then be used to alter cytokines in the middle ear, slow or stop cholesteatoma growth, or improve tympanic membrane perforation healing. Feline immunodeficiency virus (FIV), a new lentiviral vector has been found to have greater than 90% efficiency in transfecting epithelial cells. Therefore, in vivo gene therapy of middle ear mucosa cells was attempted. METHODS: Twenty microliter of 5x10(5) vectors per ml FIV carrying the gene for green fluorescence protein (GFP) was introduced into the middle ears of Sprague-Dawley rats via a bulla approach. RESULTS: Expression of the GFP gene was observed in the middle ear mucosa cells at 1 week post-inoculation indicating transfection. CONCLUSION: Gene therapy of the middle ear is feasible with a FIV-based vector. PMID- 11893455 TI - Progressive hearing loss in an infant in a neonatal intensive care unit as revealed by auditory evoked brainstem responses. AB - We herein report a case of a 14-month-old infant who revealed a progressive hearing loss by repeated auditory evoked brainstem responses (ABR) during his 1 year stay in the neonatal intensive care unit (NICU). He was born prematurely with asphyxia, hyperbilirubinemia and respiratory distress. During his 1 year stay in the NICU he was under constant mechanical ventilation. Repeated ABRs over this year initially showed normal waves but subsequently demonstrated progressive hearing impairment (HI) leading finally to no responses. Possible causes of this progressive deafness (PF) include the multiple problems of asphyxia, hyperbilirubinemia and pulmonary disorders. PMID- 11893457 TI - A rare clival and sellar fracture with pneumatocephalus. AB - We present a case of clival and sellar complex fracture produced by an indirect mechanism. This previously healthy patient had an occipital trauma followed by epistaxis. CT showed a clival and sellar fracture with pneumatocephalus. The probable fracture mechanism was contre-coup injury, linked to cerebral shock-wave transmission. This type of fracture is generally observed in the anterior part of the skull base, in a low resistance area. Severe osteoporosis probably accounted for the unusual fracture site in this patient. A mechanism of direct clival transmission is discussed, together with the usual complications of sphenoid injuries. PMID- 11893456 TI - Spontaneous external auditory canal cholesteatoma complicated by rheumatoid arthritis--case report and review of the literature. AB - A 63-year-old woman with rheumatoid arthritis sought medical assistance for dull and chronic pain in her left ear two and half years after her initial diagnostic examination. Otoscopic examination revealed that the posteroinferior wall of the bony external ear canal was eroded and that the small cavity was filled with squamous debris. The condition was diagnosed as external auditory canal cholesteatoma (EACC). The existence of EACC might suggest complications of bone disease, aging cerumen gland, or a low migratory rate of the epithelium. PMID- 11893458 TI - Conservative management of a case of cervical esophagus perforation with mediastinal abscess and bilateral pleural effusion. AB - Perforation of the cervical esophagus is a serious circumstance. Mediastinitis secondary to esophageal perforations is associated with high mortality. There is a lack of consensus on the optimal treatment for this condition. We present a case of conservative treatment in an 82-year-old woman with cervical esophagus rupture associated with mediastinal abscess and bilateral pleural effusion resulting from dilatation of a malignant esophageal stricture. Conservative treatment consisted on broad-spectrum intravenous antibiotic therapy, antireflux measures and gastrostomy was satisfactory. Treatment of the esophageal perforation should be individualized to the circumstances of each patient. Advances in antibiotic and nutritional therapy, early institution of treatment and observance of the indications, made possible a more frequent use of a conservative therapeutic approach. PMID- 11893459 TI - Arteriovenous malformation of the neck presented during delivery. AB - Vascular anomalies in the head and neck area are not infrequent and, in most cases, are congenital in origin. We present a congenital arteriovenous malformation (AVM) of the neck that manifested in a young woman during delivery. Imaging findings on ultrasonography (US), computed tomography (CT) and angiography were inconclusive and the diagnosis was reached by magnetic resonance imaging. A total excision of the mass was performed. PMID- 11893460 TI - Malignant lymphoma in the head and neck associated with benign lymphoepithelial lesion of the parotid gland. AB - Lymphoepithelial lesion is a benign lymphoproliferative disease occasionally arises in the salivary glands, but association with malignant diseases or autoimmune diseases has also been discussed. We herein present three cases of malignant lymphoma arose in the parotid gland and the lacrimal gland, following parotid surgery for benign lymphoepithelial lesion (BLEL) of the parotid gland. Two cases had mucosa associated lymphoid tissue (MALT) lymphoma in the parotid gland; one arose in the ipsilateral parotid gland as a recurrent swelling, and the other arose in the contralateral parotid gland of the previous BLEL surgery. The third case of malignant lymphoma arose in the lacrimal gland on the ipsilateral side, and the following contralateral parotid gland remained BLEL. All three patients were female, and one patient had a past history of Sjogren's syndrome and Hashimoto's disease. All three patients were treated by chemotherapy and one patient received additional radiotherapy. To follow-up lymphoproliferative diseases in the salivary glands such as BLEL, careful observation should be made on the same gland, other major salivary glands, and other organs in the head and neck, especially in females with autoimmune diseases. PMID- 11893461 TI - Hypervascular parapharyngeal schwannoma: an unusual case. AB - This report describes a 13-year-old girl who had a right pulsatile neck mass of the parapharyngeal space. We examined the patient with computerized tomography and angiography preoperatively and a heterogeneous, hypervascular mass was detected on her right neck. Intraoperative findings and the postoperative histopathologic diagnosis showed that this mass was a schwannoma that originates from cervical sympathetic chain and the superior thyroid artery supplied the mass. After careful scrutiny of English literature, this clinical manifestation is an unusual event. PMID- 11893462 TI - Metastatic testicular seminoma--a case report. AB - We present the case of a 42-year-old male who presented with a hot, tender swelling in the left supraclavicular fossa. He was pyrexial on presentation with a mildly elevated leucocyte count of 12.4x10(9)/l. Clinical examination, including full ear, nose and throat assessment, proved unremarkable. The medical history revealed that 2 years earlier the patient had been diagnosed with a testicular seminoma for which he underwent a right inguinal orchidectomy and abdominal radiotherapy. CT scan highlighted a 6 cm para-laryngeal mass, of mixed attenuation, with an adjacent region of inflammation. Overall appearance was suggestive of an infective mass. Ultrasound-guided fine needle aspiration cytology revealed a metastatic seminomatous deposit. Imaging of the chest and abdomen revealed this as the only site of metastasis. He is currently undergoing chemotherapy, and is responding well. We review the pathology of testicular seminoma. This case highlights the myriad of pathologies that may present as a lump in the neck. PMID- 11893463 TI - Have you noticed that surgery has changed?. The laparoscopic approach to the spleen. PMID- 11893464 TI - Current treatment strategies for multiple myeloma. AB - Until 1990, the melphalan-prednisone regimen was the standard treatment for multiple myeloma (MM). The role of alpha-interferon still remains controversial, both in induction therapy and in maintenance therapy. Over the last 10 years, there has been considerable improvement in the treatment of MM. In patients under 65 years of age, high-dose therapy with autografting has clearly demonstrated an advantage over conventional treatment. Bisphosphonates have proved very useful in reducing skeletal events. More recently, an old drug, thalidomide, has shown surprising efficacy in patients with advanced MM. Future trends include the extension of high-dose therapy to older patients and the use of immunotherapy in induction and/or maintenance therapy. PMID- 11893465 TI - Beta adrenergic blockers in chronic congestive cardiac failure: a call for action. AB - Antagonism of adverse neuro-endocrine responses is the current paradigm by which chronic congestive cardiac failure is treated. Several recent large-scale randomized, controlled trials have confirmed the benefits of beta-adrenergic blocking agents in this regard. In light of the present heart failure 'epidemic', appropriate organization of services is mandatory to ensure that as many patients as possible can benefit from this life-saving therapy. PMID- 11893466 TI - Prevention of tumor spread by matrix metalloproteinase-9 inhibition: old drugs, new concept. AB - In the following review we will discuss some familiar drugs with potential use as adjuvant anti-neoplastic agents. We will highlight their unfamiliar property of being able to inhibit matrix metalloproteinase-9 activity, which has recently been proven to play a key role in cancer spread. These drugs' high-safety profile may allow clinicians to construct new anti-cancer protocols that may prove to be less toxic alternatives to those commonly used. PMID- 11893467 TI - Oral clarithromycin enhances gallbladder emptying induced by a mixed meal in healthy subjects. AB - Background: In humans, erythromycin has been demonstrated to accelerate gallbladder emptying due to its motilin-like effects on the gastrointestinal tract. Recently, it was shown that clarithromycin, another macrolide, used for the eradication of Helicobacter pylori infection, also stimulated gastrointestinal motility in the fasting state. We conducted a comparative study on the effects of a single oral dose of clarithromycin and of erythromycin on gallbladder emptying in healthy subjects. Methods: Gallbladder emptying variables (residual volume, ejection fraction, area under emptying curve) were measured by ultrasound in 21 healthy subjects (11 males, 10 females, mean age 42.5+/-10.6 years). A test meal (14 g fat, 425 kcal) was ingested 30 min after a single oral dose (500 mg) of either clarithromycin or erythromycin, and the measurements were repeated the following day with the other drug (cross-over double-blind study). A control group consisting of 12 subjects (seven males, five females, mean age 50.7+/-8.2 years) was used to evaluate gallbladder emptying following the same test meal without drug administration. Differences between groups were analyzed using two-tailed Student's t-test for unpaired observations. Results: Gallbladder emptying at 60, 75, and 90 min was greater after erythromycin (P<0.05 at 90 min) and clarithromycin than it was in controls. The ejection fraction was significantly greater after clarithromycin (76.5%) and erythromycin (79.7%) than it was in controls. Gallbladder refilling occurred earlier after clarithromycin than after erythromycin. Conclusions: The prokinetic effect of clarithromycin on the gallbladder appears to be of similar amplitude but of shorter duration than that of erythromycin. PMID- 11893468 TI - Insulin resistance and metabolic parameters in type 2 diabetic patients. AB - Background: The importance of insulin resistance in type 2 diabetes mellitus has been generally accepted. Only very few data about the degree of insulin resistance in a representative group of type 2 diabetic patients are available. The aim of this study was to ascertain the degree of insulin resistance and its relation to metabolic parameters in type 2 diabetic patients. Methods: We studied 96 type 2 diabetic patients. The inclusion criteria were type 2 diabetes according to WHO criteria and HbA(1c) between 6.8% and 10.5%. Insulin resistance was estimated in a euglycemic hyperinsulinemic clamp. Blood parameters like lipids, insulin, glucose, fatty acids, and leukocytes were also studied. Results: The insulin sensitivity of 71 of the type 2 diabetic patients was markedly lower than that of the controls. Twenty-five diabetic patients had an M(c) value within the range of the controls. The M(c) values, as a measure of insulin resistance, of the diabetic patients were between 0.3 and 5.2 mg/(kg min insulin), whereas the M(c) range of the controls was from 2.6 to 10.8 mg/(kg min insulin). Conclusions: Approximately 75% of the type 2 diabetic patients were insulin resistant. Hence, type 2 diabetes mellitus was not equivalent to insulin resistance in every case. PMID- 11893469 TI - Selected cytokines and soluble forms of cytokine receptors in coronary artery disease. AB - Background: Cytokines may play a role in the pathogenesis of atherosclerosis and coronary artery disease (CAD). Methods: We examined serum concentrations of selected pro- (TNFalpha, IL-2) and anti-inflammatory (IL-10) cytokines, and soluble forms of TNF receptors (sTNFR 1 and sTNFR 2) by ELISA in 45 patients with stable exertional angina (group 1), 32 patients with unstable angina (group 2), and 20 healthy subjects (group C). Results: Serum concentrations of both TNFalpha (group 1, 18.3; group 2, 17.2 pg/ml; P<0.001) and IL-10 (group 1, 46.1; group 2, 41.5 pg/ml; P<0.05) were significantly higher in patients with CAD than in group C (8.3 and 14.3 pg/ml, respectively). sTNFR 1 serum level was higher in group 1 (1399.6; P<0.05) than in healthy volunteers (1093.9 pg/ml). In turn, the serum level of IL-2 was significantly higher in unstable patients than it was in groups 1 and C (89.4, 59.8 and 52.8 pg/ml, respectively). In group 1, both TNFalpha and IL-2 correlated with serum lipids. Conclusions: Patients with CAD, irrespective of the form of the disease, have higher serum levels of pro- and anti inflammatory cytokines than control subjects. Increased concentrations of IL-2 in unstable angina may suggest additional immunologic activation. The pro inflammatory cytokine levels seem to be related to lipid disturbances. PMID- 11893470 TI - Gallbladder and gastric emptying: relationship to cholecystokininemia in diabetics. AB - Background: Impaired gastrointestinal and gallbladder motility, as a complication of long-lasting diabetes mellitus, has been ascribed to the possible development of autonomic neuropathy, although the intervention of hormonal factors may not completely be excluded. In this regard, cholecystokinin (CCK), a gut hormone known to regulate pancreatic exocrine secretion, gallbladder contraction, and bowel motility in response to a meal, is impaired in patients with diabetes mellitus. This prompted us to evaluate the relationship between the plasma levels of CCK and gallbladder and gastric emptying in neuropathy-free diabetic patients treated with insulin (group A) or with oral hypoglycemic agents (group B) under basal conditions and in response to a standard test meal. Methods: Plasma CCK was measured by radioimmunoassay. Gastric and gallbladder emptying were evaluated ultrasonographically. Results: Plasma CCK levels were significantly lower in both groups of diabetics than in healthy controls during a fast and in response to a standard meal. However, meal ingestion was able to evoke a pattern of CCK response in both groups of diabetic patients similar to that seen in controls. Fasting gallbladder volume was higher in patients with diabetes than in controls, whereas the percentage of emptying was lower in patients of both groups. Gastric final emptying time was significantly longer in both groups of diabetics than in controls. Conclusion: This study shows that patients with diabetes have lower plasma levels of CCK, which may explain their relatively hypotonic gallbladder and reduced gastric motility. PMID- 11893471 TI - Sarcoidosis associated with interferon-alpha therapy for chronic hepatitis B. AB - Sarcoidosis is one of the possible rare complications of interferon-alpha (IFN alpha) therapy. Only a few reports have been published on this disease, and these have been associated with the treatment of malignant diseases, essential thrombocytosis, and chronic hepatitis C. We report on a 64-year-old man with chronic hepatitis B (HBsAg, HBeAg, HBV DNA-positive) who was treated with recombinant IFN-alpha-2b (5 MU three times weekly) for 28 weeks. Tolerance to treatment was very good; only a mild flu-like syndrome appeared. Twelve months after completing the therapy, a chest X-ray was performed that revealed bilateral hilar masses, and high-resolution computed tomography (HRCT) of the chest indicated the presence of lymphadenopathy of the anterior and middle mediastinum. Therefore, a right-sided thoracoscopy was performed with excision of a 27-mm lymph node and a histological diagnosis of sarcoidosis was made. No medication for sarcoidosis was indicated. Complete normalization of mediastinal lymphadenopathy (verified on HRCT and chest X-ray) was confirmed 1 year following the thoracoscopy. To our knowledge, this is the first case wherein occurrence of sarcoidosis in a chronic hepatitis B patient treated with IFN-alpha is described. We suppose that IFN-alpha, as a potent stimulator of T-helper 1 (Th1) immune responses, may trigger the compartmentalized Th1 reaction that has been shown to take place in sarcoidosis. PMID- 11893472 TI - High CA 19-9 levels in benign biliary tract diseases. Report of four cases and review of the literature. AB - For years, CA 19-9 has been proposed as a marker for epithelial-type gastrointestinal cancers, even though it is well known that its diagnostic specificity is low. Here we describe cases of extremely high CA 19-9 levels in benign biliary tract diseases. The first case involved a 77-year-old male patient with choledocholithiasis and jaundice who was found to have CA 19-9 levels of 98,628 UI/ml. The second case was a 63-year-old male patient with autoimmune cholangitis and a CA 19-9 level of 250 IU/ml. The third case was a 74-year-old male patient with cholelithiasis and choledocholithiasis who developed acute cholangitis. CA 19-9 levels were elevated to 14,950 UI/ml during the episode. The fourth case involved a 73-year-old man with biliary colic and jaundice following an acute open cholecystectomy procedure who had a transient 100-fold increase in CA 19-9 (2230 IU/ml). PMID- 11893473 TI - Sensitivity of the criteria used to diagnose adult still's disease in internal medicine practice. a study of 17 cases. AB - Our aim was to compare the sensitivity of various diagnostic criteria for adult Still's disease in 17 patients with established adult Still's disease who were followed in a department of internal medicine over a mean period of 7 years. The median age of the 17 patients was 27 years and the sex ratio M:F was 1:4. The patients had essentially systemic manifestations with fever (n=14), inflammation and leukocytosis (n=16), and moderate liver dysfunction (n=13) that disappeared after a mean of 21 days (range 8-48 days). In the first week of hospitalization, the sensitivities of the criteria proposed by Yamaguchi, Reginato, and Kahn were 94, 18, and 23%, respectively. One month later, the sensitivity was 100% for Yamaguchi's diagnostic criteria versus only 88% for Reginato's and Kahn's criteria. This study confirms that Yamaguchi's diagnostic criteria are more sensitive and are met earlier than Reginato's and Kahn's criteria in patients followed in internal medicine. PMID- 11893474 TI - Aspergillus spondylodiskitis in a patient with chronic obstructive pulmonary disease. AB - Invasive aspergillosis is a well-known complication in immunocompromised patients. There are only a few reports of invasive aspergillosis in non immunocompromised patients. We describe a 72-year-old female patient with clinical signs of spondylodiskitis occurring 4 months after what had appeared to be successful treatment of pulmonary aspergillosis. The patient used inhalation corticosteroids on a daily basis because of chronic obstructive pulmonary disease (COPD). Spondylodiskitis of the intervertebral disc Th11 and Th12 with involvement of both adjacent vertebral bodies was confirmed by magnetic resonance imaging. Histopathological examination revealed the presence of septate hyphae, indicative of Aspergillus species. Subsequently, evidence of Aspergillus spondylodiskitis was obtained by amplification of Aspergillus-DNA with a specific polymerase chain reaction method. Aspergillus spondylodiskitis after pulmonary aspergillosis is only very rarely encountered. Patients with COPD, managed with short-term courses of systemic corticosteroids or with high-dose corticosteroid inhalation therapies, are considered non-immunocompromised but might be at risk of developing invasive aspergillosis. PMID- 11893475 TI - Upper gastrointestinal evaluation of asymptomatic patients with iron-deficiency anemia after a negative colonoscopy. PMID- 11893476 TI - The authors reply. PMID- 11893477 TI - Soy isoflavones' osteoprotective role in postmenopausal women: mechanism of action. AB - Ovarian hormone deficiency is a major risk factor for osteoporosis. Current therapies emphasize the use of antiresorptive agents, such as estrogen, calcitonin, and bisphosphonates. These therapies are associated with certain risks and side effects making compliance a major obstacle. Recent findings suggest that a class of synthetic and naturally occurring compounds, selective estrogen receptor modulators, e.g. raloxifene and soy isoflavones can offer attractive alternatives. Evidence for bone-sparing effects of isoflavones relies mainly on animal findings supported by a limited number of human studies. These observations suggest that isoflavones exert their effects on bone by stimulating bone formation and at the same time suppressing bone resorption. However, the precise osteoprotective mechanism of isoflavones remains uncertain and awaiting further clarification. From a clinical point of view, larger and longer duration studies are warranted to enable us to draw clear conclusions in regards to the role of isoflavones on bone. PMID- 11893478 TI - Effects of copper and ceruloplasmin on iron transport in the Caco 2 cell intestinal model. AB - Previous studies have implicated copper proteins, including ceruloplasmin, in intestinal iron transport. Polarized Caco2 cells with tight junctions were used to examine the possibilities that (a) ceruloplasmin promotes iron absorption by enhancing release at the basolateral cell surface and (b) copper deficiency reduces intestinal iron transport. Iron uptake and overall transport were followed for 90 min with 1 &mgr;M 59Fe(II) applied to the apical surface of Caco2 cell monolayers. Apotransferrin (38 &mgr;M) was in the basolateral chamber. Induction of iron deficiency with desferrioxamine (100 &mgr;M; 18 h) markedly increased uptake and overall transport of iron. Uptake increased from about 20% to about 65% of dose, and overall 59Fe transport from <1% to 60% of dose. On the basis of actual iron released into the basal chamber (measured with bathophenanthroline), transport increased 8-fold. Desferrioxamine pretreatment reduced cellular Fe by 55%. The addition of freshly isolated, enzymatically active human ceruloplasmin to the basolateral chamber during absorption had no effect on uptake or transport of iron by the cells. Unexpectedly, pretreatment with three different chelators of copper (18 h), which reduced cellular levels about 40%, more than doubled iron uptake and raised overall transport to 20%. This was so, whether or not cells were also made iron deficient with desferrioxamine. Acute addition of 1 &mgr;M Cu(II) to the apical chamber had no significant effect upon iron uptake, retention, or transport in iron deficient or normal cells, in the presence of absence of ascorbate. We conclude that intestinal absorption of Fe(II) is unlikely to depend upon plasma ceruloplasmin, and that cuproproteins involved in this form of iron transport must be binding copper tightly. PMID- 11893479 TI - Improved growth of lipoprotein lipase deficient kittens by feeding a low-fat, highly digestible diet small star, filled. AB - Adult domestic cats homozygous with a naturally occurring Gly412Arg LPL gene mutation are good models for the study of LPL deficiency. Previous studies report that homozygous LPL deficient kittens have reduced growth rates and develop subnormal body fat mass. It was hypothesized in the present study that homozygote kittens would have normal growth if provided a standard low fat, highly digestible diet at weaning and that their body fat would be increased by provision of a diet high in protein. When fed a nutritionally complete, 10% fat, purified or commercial extruded diet, the body weights of homozygous (n = 24), heterozygous (n = 25) and normal (n = 16) kittens were determined at birth, 2, 3, 4, 6, 12 and 18 weeks of age. Male homozygote kittens from homozygote dams had reduced weight gains (p < 0.05) compared to normal males at 2, 3 and 4 weeks. Female heterozygotes and homozygotes from homozygote and heterozygote dams had reduced weight gains (p < 0.05) compared to normal females at 2, 3, 4 and 6 weeks. By 6 weeks for males and 18 weeks for females, genotype related differences in weight gain were not observed. At 30 weeks, homozygotes and heterozygotes were given either a 60 or 30% (dry matter) protein diet for two months. As indicated by deuterium dilution estimation of body composition, cats eating the 30% protein diet (n = 12) tended to have a lower increase in lean body mass (p = 0.057) and a greater increase in fat mass (p = 0.092) compared to cats eating the 60% protein diet (n = 12). Increase in lean body mass among homozygotes tended to be not as great as that observed in heterozygotes (p = 0.057). Poor postweaning gains previously reported in homozygotes probably reflected inappropriate selection of diet for this genotype. The high protein diet increased the rate of lean body mass development but not body fat mass. PMID- 11893480 TI - The effect of chitosan and other polycations on tight junction permeability in the human intestinal Caco-2 cell line(1). AB - Chitosan is a polycationic compound widely employed as dietary supplement and also present in pharmaceutical preparations. Although it has been approved for human consumption, its possible side effects have not been widely investigated and the available data in the literature are still controversial. Several polycationic substances have been shown to affect tight junction permeability in epithelial cell models in vitro. In this study we have compared the effects of chitosan and other polycations (polyethylenimine, poly-L-lysines of different molecular weights) on the integrity of tight junctions and of the actin cytoskeleton in the human intestinal Caco-2 cell line. We have measured trans epithelial electrical resistance and paracellular passage of the extracellular marker inulin, and we have localized F-actin and tight junctional proteins (ZO1 and occludin) in cell monolayers treated with various concentrations of each polycation. Fluorescent poly-L-lysines were also employed to determine their association with the cell monolayer. Our results indicate that all polycations investigated are able to induce a reversible increase in tight junction permeability. This effect is concentration and energy dependent, affected by the extracellular concentration of divalent cations (calcium, magnesium and manganese) and it is associated with morphological changes in the F-actin cytoskeleton, as well as in the localization of tight junctional proteins. Chitosan, in particular, was the only cationic polymer that displayed an irreversible effect on tight junctions at the highest concentration tested (0.01%). These results indicate that oral ingestion of chitosan may have more widespread health effects by altering intestinal barrier function, thus allowing the entrance into the circulation of potentially toxic and/or allergenic substances. PMID- 11893481 TI - HPLC and GC/MS determination of deuterated vitamin K (phylloquinone) in human serum after ingestion of deuterium-labeled broccoli. AB - The ability to intrinsically label plant constituents with stable isotopes has the potential to advance the study of vitamin K-absorption and metabolism in humans. Broccoli, a primary food source of phylloquinone (VK-1), was grown hydroponically using 31 atom % deuterium oxide in order to label VK-1 within the food matrix. Deuterium-labeled broccoli (115 g; 168 &mgr;g VK-1) was fed to one male subject in a single serving. Multiple serum samples were drawn throughout the subsequent 24-hr period. Reversed-phase HPLC was used to extract and purify VK-1 in both broccoli and serum. Ion abundances of the deuterium-labeled and unlabeled (endogenous) VK-1 were determined using GC/MS in negative chemical ionization mode. No sample derivatization was required. Endogenous VK-1 produced isotopomers from m/z 450 to m/z 453. The labeled VK-1 isotopomers in the broccoli were from m/z 452 to m/z 467, with the most abundant isotopomer being m/z 458 (14.1% of total labeled VK-1). The GC/MS chromatograms from serum revealed both endogenous VK-1 and VK-1 derived from the deuterium-labeled broccoli. The profile of labeled VK-1 isotopomers in serum was identical to the VK-1 isotopomer profile in labeled broccoli, indicating that no deuterium was lost due to exchange either in the body or in sample preparation. At 4 hr following broccoli intake, there was an 81.1% enrichment of phylloquinone in serum; labeled VK-1 was no longer detectable in serum at 24 hr. Use of isotope labeled vegetables enables one to discriminate exogenous intake of VK-1 from endogenous pools and ultimately to determine bioavailability of VK-1 from foods. PMID- 11893482 TI - Effects of stabilized rice bran, its soluble and fiber fractions on blood glucose levels and serum lipid parameters in humans with diabetes mellitus Types I and II. AB - Stabilized rice bran (SRB), a source of complex carbohydrates, tocols, gamma oryzanols, and polyphenols, was treated with carbohydrases and heat to yield two fractions, rice bran water solubles (RBWS), and rice bran fiber concentrates (RBFC). Stabilized rice bran and its fractions were fed for 60 days to insulin dependent and noninsulin-dependent diabetes mellitus (IDDM = Type I and NIDDM = Type II) subjects to determine possible effects on serum hemoglobin, carbohydrate and lipid parameters. The Type I subjects (n = 22, 26, and 20) fed Stabilized rice bran, rice bran water solubles, and rice bran fiber concentrates plus AHA Step-1 diet reduced glycosylated hemoglobin 1%, 11%, and 10%, respectively. The fasting serum glucose levels were also reduced significantly (P < 0.01) with stabilized rice bran (9%), rice bran water solubles (29%), and rice bran fiber concentrates (19%).The Type II subjects (n = 31, and 26) fed rice bran water solubles and rice bran fiber concentrates plus AHA Step-1 diet had decreased levels of glycosylated hemoglobin (15% and 11%) and fasting glucose (33% and 22%; P < 0.001), respectively. Serum insulin levels were increased (4%) with rice bran water solubles in both types of diabetes. The reduction of glycosylated hemoglobin and a slight increase in insulin levels indicate that consumption of rice bran water solubles can control blood glucose levels in human diabetes. Serum total cholesterol, LDL-cholesterol, apolipoprotein B, and triglycerides levels were reduced with rice bran fiber concentrates in the Type I (10, 16, 10, 7%) and Type II groups (12, 15, 10, 8%), respectively. These results indicate that rice bran water solubles significantly reduces hyperglycemia (P < 0.01), whereas rice bran fiber concentrates reduces hyperlipidemia (P < 0.05) in both types of diabetes. Therefore, these natural products can be used as nutritional supplements for the control of both types of diabetes mellitus in humans. PMID- 11893483 TI - Chromosome and expression mechanisms: a year dominated by histone modifications, transitory and remembered. PMID- 11893484 TI - Deciphering gene expression regulatory networks. AB - In the past year, great strides have been made in our understanding of the regulatory networks that control gene expression in the model eukaryote Saccharomyces cerevisiae. The development and use of a number of genomic tools, including genome-wide location and expression analysis, has fueled this progress. In addition, a variety of computational algorithms have been devised to mine genomic sequence for conserved regulatory motifs in co-regulated genes. The recent description of the genetic network controlling the cell cycle illustrates the tremendous potential of these approaches for deciphering gene expression regulatory networks in eukaryotic cells. PMID- 11893485 TI - Protein dynamics in the nuclear compartment. AB - The classic view of a transcriptional initiation complex is that of an assembly of factors with many protein-protein contacts, leading to a multi-component complex whose existence is a result of the stabilizing influence of the many intermolecular interactions. Recent findings from protein mobility experiments in living cells indicate that many kinds of nuclear factors move rapidly and exchange quickly with multiple targets. Two countervailing views of factor/regulatory site interactions emerge from the current literature. PMID- 11893486 TI - Histone modifications in transcriptional regulation. AB - Covalent modifications of the amino termini of the core histones in nucleosomes have important roles in gene regulation. Research in the past two years reveals these modifications to consist of phosphorylation, methylation and ubiquitination, in addition to the better-characterized acetylation. This multiplicity of modifications, and their occurrence in patterns and dependent sequences, argues persuasively for the existence of a histone code. PMID- 11893487 TI - Coactivators in transcription initiation: here are your orders. AB - Coactivators are diverse and multifunctional proteins that act downstream of DNA binding activators to stimulate transcription. Recent studies elucidate the temporal sequence in which coactivators are recruited to target promoters, and how their enzymatic properties and molecular interactions culminate in transcriptional initiation. PMID- 11893488 TI - Chromatin elongation factors. AB - As RNA polymerase II leaves a gene promoter to transcribe the coding region, it faces a major obstacle - nucleosomes tightly wrapped into chromatin. Mechanisms to deal with this obstacle clearly exist in cells, as transcription through chromatin is very efficient in vivo, whereas nucleosomal templates pose a considerable problem for polymerase progression in reconstituted in vitro systems. Advances in our understanding of transcriptional elongation through chromatin have been made possible recently by the identification of several accessory factors that assist polymerase in the process. Insights into the function of these factors have been gained by a combination of yeast genetics and biochemical studies in mammalian systems. PMID- 11893489 TI - Histone H2A variants H2AX and H2AZ. AB - Two of the nucleosomal histone families, H3 and H2A, have highly conserved variants with specialized functions. Recent studies have begun to elucidate the roles of two of the H2A variants, H2AX and H2AZ. H2AX is phosphorylated on a serine four residues from the carboxyl terminus in response to the introduction of DNA double-strand breaks, whether these breaks are a result of environmental insult, metabolic mistake, or programmed process. H2AZ appears to alter nucleosome stability, is partially redundant with nucleosome remodeling complexes, and is involved in transcriptional control. PMID- 11893490 TI - ChIPs of the beta-globin locus: unraveling gene regulation within an active domain. AB - Recent studies of beta-globin gene expression have concentrated on the analysis of factor binding and chromatin structure within the endogenous locus. These studies have more precisely defined the extent and nature of the active chromosomal domain and the elements that organize it. Surprisingly, the beta globin locus control region (LCR), although critical for high-level gene expression, plays little role in the overall architecture of the active locus. Analysis of the effects of targeted deletion of the beta-globin LCR, along with emerging knowledge of the behavior of the erythroid transcription factor NF-E2, leads to a new perspective on factor binding and LCR function. PMID- 11893492 TI - Breaking through to the other side: silencers and barriers. AB - The establishment and restriction of transcriptionally inactive regions in the nucleus is mediated by silencer and barrier elements. Silencer-bound proteins recruit additional factors to establish the silenced domain during the S-phase of the cell cycle but, contrary to previous models, DNA replication is not a pre requisite for the establishment. Characteristically, silenced domains contain hypoacetylated histones and recent data have identified residue-specific methylation of histone H3 as an additional signature that distinguishes active regions from inactive ones. Peaks of acetylated histones demarcate the boundaries between these regions and recruitment of HAT activities provides a mechanism to restrict the spread of heterochromatin. PMID- 11893491 TI - Heterochromatin: new possibilities for the inheritance of structure. AB - Significant portions of the eukaryotic genome are heterochromatic, made up largely of repetitious sequences and possessing a distinctive chromatin structure associated with gene silencing. New insights into the form of packaging, the associated histone modifications, and the associated nonhistone chromosomal proteins of heterochromatin have suggested a mechanism for providing an epigenetic mark that allows this distinctive chromatin structure to be maintained following replication and to spread within a given domain. PMID- 11893493 TI - Gene silencing, cell fate and nuclear organisation. AB - Although advances in molecular biology have allowed us to identify and describe many of the events associated with turning genes on, much less attention has generally been focussed on the related process of gene silencing. This is surprising as heritable gene inactivation plays an important role in determining cell lineage fates during development and defining their temporal order. Recent advances in the area of chromatin and chromosome organisation may have an impact on our understanding of cellular differentiation. PMID- 11893494 TI - Histone methylation in transcriptional control. AB - Over the past year or so, methylation of histones has come to be recognised as a major player in the regulation of gene activity. This notion follows the discovery of lysine and arginine methyltransferases and proteins that recognise the methyl-lysine 'mark' on histones. Methylated histones have been implicated in heterochromatic repression, promoter regulation and the propagation of a repressed state via DNA methylation. PMID- 11893495 TI - Programming off and on states in chromatin: mechanisms of Polycomb and trithorax group complexes. AB - Polycomb and trithorax group proteins are evolutionarily conserved chromatin components that maintain stable states of gene expression. Recent studies have identified and characterized several multiprotein complexes containing these transcriptional regulators. Advances in understanding molecular activities of these complexes in vitro, and functional domains present in their subunits, suggest that they control transcription through multistep mechanisms that involve nucleosome modification, chromatin remodeling, and interaction with general transcription factors. PMID- 11893496 TI - X-chromosome inactivation and the search for chromosome-wide silencers. AB - X-chromosome inactivation leads to divergent fates for two homologous chromosomes. Whether one X remains active or becomes silenced depends on the activity of Xist, a gene expressed only from the inactive X and whose RNA product 'paints' the X in cis. Recent work argues that Xist RNA itself is the acting agent for initiating the silencing step. Xist RNA contains separable domains for RNA localization and chromosome silencing. While no Xist RNA-interacting factors have been identified, a growing collection of chromatin alterations have been identified on the inactive X, including variant histone H2A composition and histone H3 methylation. Some or all of these changes may be critical for chromosome-wide silencing. As none of the silencing proteins identified so far is unique to X chromosome inactivation, the specificity must partly reside in Xist RNA whose spread along the X orchestrates general silencing factors for this specific task. PMID- 11893497 TI - RNAi: nature abhors a double-strand. AB - In organisms as diverse as nematodes, trypanosomes, plants, and fungi, double stranded RNA triggers the destruction of homologous mRNAs, a phenomenon known as RNA interference. RNA interference begins with the transformation of the double stranded RNA into small RNAs that then guide a protein nuclease to destroy their mRNA targets. PMID- 11893498 TI - Biotechnologies and therapeutics: chromatin as a target. AB - As alterations in gene expression underlie a considerable proportion of human diseases, correcting such aberrant transcription in vivo is expected to provide therapeutic benefit to the patient. Attempts to control endogenous mammalian genes, however, face a significant obstacle in the form of chromatin. Aberrant gene repression can be alleviated by using small-molecule inhibitors that exert nucleus-wide effects on chromatin-based repressors. Genome-wide chromatin remodeling also occurs during cloning via nuclear transfer, and causes the deregulation of epigenetically controlled genes. Regulation of genes in vivo can be accomplished via the use of designed transcription factors - these result from a fusion of a designed DNA-binding domain based on the zinc finger protein motif to a functional domain of choice. PMID- 11893499 TI - DNA replication and chromatin. AB - The study of DNA replication in eukaryotic chromosomes has revealed a multitude of different regulatory levels. Nuclear and chromosomal location as well as chromatin structure may affect the activity of replication origins and their modulation during development. PMID- 11893500 TI - Proteomics of the nucleolus: more proteins, more functions? PMID- 11893501 TI - AMOP, a protein module alternatively spliced in cancer cells. AB - This article describes a new extracellular domain--AMOP, for adhesion-associated domain in MUC4 and other proteins. This domain occurs in putative cell adhesion molecules and in some splice variants of MUC4. MUC4 splice variants are overexpressed in several tumours; in particular, they are highly expressed in pancreatic carcinomas but not in normal pancreas. The presence of AMOP in cell adhesion molecules could be indicative of a role for this domain in adhesion. PMID- 11893502 TI - A second catalytic domain in the Elp3 histone acetyltransferases: a candidate for histone demethylase activity? AB - A new subfamily of two-domain histone acetyltransferases (HATs) related to Elp3 has been identified. In addition to a HAT domain in the C terminus, these proteins have an N-terminal domain similar to the catalytic domain of S adenosylmethionine radical enzymes. Two-domain organization is preserved in evolution, suggesting that both enzymatic activities are functionally or mechanistically coupled and directed towards highly conserved substrates. The functional implications of this similarity and a possible role for Elp3-related proteins as histone demethylases are discussed. PMID- 11893508 TI - Protein recognition by cell surface receptors: physiological receptors versus virus interactions. AB - Protein-protein recognition is a major kind of receptor-ligand interaction: a living cell receives external signals to adapt to the environment through cell surface receptors. On opposing cell surfaces, such recognition bears distinct features: it is a multivalent, reversible and avidity-driven process. The affinity between each individual contacting pair is low. Viruses might take advantage of this low affinity to invade a host cell by evolving a stronger binding affinity to the surface receptors than that associated with physiological ligands. Structural data appear to support this notion. PMID- 11893509 TI - The MUC family: an obituary. AB - Mucins are glycoproteins that are common on the surfaces of many epithelial cells; they are deemed to mediate many interactions between these cells and their milieu. Several of these mucins form the mucus layer that is found in many hollow organs. The biophysical properties of mucins are related to their extensive O linked glycosylation rather than directly to their polypeptide sequences. Despite the frequent absence of sequence homology, many human genes encoding mucins have been named MUC followed by a number, unjustly suggesting the existence of one large gene family. In this article, it is suggested that the mucin genes be renamed according to their sequence homologies. PMID- 11893510 TI - Circular proteins--no end in sight. AB - Circular proteins are a recently discovered phenomenon. They presumably evolved to confer advantages over ancestral linear proteins while maintaining the intrinsic biological functions of those proteins. In general, these advantages include a reduced sensitivity to proteolytic cleavage and enhanced stability. In one remarkable family of circular proteins, the cyclotides, the cyclic backbone is additionally braced by a knotted arrangement of disulfide bonds that confers additional stability and topological complexity upon the family. This article describes the discovery, structure, function and biosynthesis of the currently known circular proteins. The discovery of naturally occurring circular proteins in the past few years has been complemented by new chemical and biochemical methods to make synthetic circular proteins; these are also briefly described. PMID- 11893511 TI - Conservation of amino acid transporters in fungi, plants and animals. AB - When comparing the transporters of three completely sequenced eukaryotic genomes- Saccharomyces cerevisiae, Arabidopsis thaliana and Homo sapiens--transporter types can be distinguished according to phylogeny, substrate spectrum, transport mechanism and cell specificity. The known amino acid transporters belong to five different superfamilies. Two preferentially Na(+)-coupled transporter superfamilies are not represented in the yeast and Arabidopsis genomes, whereas the other three groups, which often function as H(+)-coupled systems, have members in all investigated genomes. Additional superfamilies exist for organellar transport, including mitochondrial and plastidic carriers. When used in combination with phylogenetic analyses, functional comparison might aid our prediction of physiological functions for related but uncharacterized open reading frames. PMID- 11893512 TI - Iron hydrogenases--ancient enzymes in modern eukaryotes. AB - The distribution of [Fe]-hydrogenases was once thought to be limited to a small number of bacteria and a few peculiar hydrogen-producing anaerobic eukaryotes. However, it is now clear that [Fe]-hydrogenases are more widely distributed among eukaryotes than reports of hydrogen production have suggested. Indeed, genes bearing the hallmark signatures of [Fe]-hydrogenases are found both in our own genome and in the genomes of other higher eukaryotes. At present, the functions of most of these new proteins remain unknown; it is not even known whether they can all make hydrogen. Radical new hypotheses have suggested that hydrogenases played a key role in the formation of the eukaryotic cell. These unique enzymes have thus moved from the margins of eukaryotic biology to become the focus of intense speculation and interest. This article summarizes current knowledge of their distribution, evolution and biochemistry. PMID- 11893513 TI - Mechanism of the F(1)F(0)-type ATP synthase, a biological rotary motor. AB - The F(1)F(0)-type ATP synthase is a key enzyme in cellular energy interconversion. During ATP synthesis, this large protein complex uses a proton gradient and the associated membrane potential to synthesize ATP. It can also reverse and hydrolyze ATP to generate a proton gradient. The structure of this enzyme in different functional forms is now being rapidly elucidated. The emerging consensus is that the enzyme is constructed as two rotary motors, one in the F(1) part that links catalytic site events with movements of an internal rotor, and the other in the F(0) part, linking proton translocation to movements of this F(0) rotor. Although both motors can work separately, they must be connected together to interconvert energy. Evidence for the function of the rotary motor, from structural, genetic and biophysical studies, is reviewed here, and some uncertainties and remaining mysteries of the enzyme mechanism are also discussed. PMID- 11893514 TI - Getting the most from PSI-BLAST. AB - Most biologists now conduct sequence searches as a matter of course. But how do we know that a relationship predicted by a homology search is a true, rather than false, hit with the same score? Many biologists design their own experiments with exquisite care yet still assume that results from programs with more than 20 adjustable parameters are 100% reliable. This article explains some of the key steps in getting the most from PSI-Blast, one of the most popular and powerful homology search programs currently available. PMID- 11893516 TI - More than scratching the surface: mitogen receptors as transcription factors? PMID- 11893517 TI - Cortisol, 11 beta-hydroxysteroid dehydrogenase type 1 and central obesity. PMID- 11893518 TI - Current approaches and new advances in endocrine hypertension. PMID- 11893522 TI - New clues about vitamin D functions in the nervous system. AB - Accumulating data have provided evidence that 1 alpha,25 dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] is involved in brain function. Thus, the nuclear receptor for 1,25-(OH)(2)D(3) has been localized in neurons and glial cells. Genes encoding the enzymes involved in the metabolism of this hormone are also expressed in brain cells. The reported biological effects of 1,25-(OH)(2)D(3) in the nervous system include the biosynthesis of neurotrophic factors and at least one enzyme involved in neurotransmitter synthesis. 1,25-(OH)(2)D(3) can also inhibit the synthesis of inducible nitric oxide synthase and increase glutathione levels, suggesting a role for the hormone in brain detoxification pathways. Neuroprotective and immunomodulatory effects of this hormone have been described in several experimental models, indicating the potential value of 1,25 (OH)(2)D(3) pharmacological analogs in neurodegenerative and neuroimmune diseases. In addition, 1,25-(OH)(2)D(3) induces glioma cell death, making the hormone of potential interest in the management of brain tumors. These results reveal previously unsuspected roles for 1,25-(OH)(2)D(3) in brain function and suggest possible areas of future research. PMID- 11893523 TI - Sex with two SOX on: SRY and SOX9 in testis development. AB - Although gonads are not required for development or survival, defects in gonadal development undoubtedly have a profound influence on affected individuals. Recent complementary studies in the fields of cytology, biochemistry and molecular genetics have revealed that normal gonad development involves an exquisitely regulated network of gene expression and protein-protein interactions. The initial event of gonadogenesis, in both males and females, involves the formation of a bipotential primordium. A Y chromosome then activates the male-specific pathway. The demonstration that mutations in the SOX proteins, SRY and SOX9, are responsible for disorders associated with male-to-female sex reversal showed dramatically that SRY and SOX9 have an essential role in male sex differentiation. This was emphasized when it was shown that female mice carrying transgenes that encode these proteins developed as males. SRY and SOX9 proteins have been characterized extensively and aspects of their function and regulation are now known. PMID- 11893524 TI - GnRH neuronal development: insights into hypogonadotrophic hypogonadism. AB - Pulsatile secretion of the hypothalamic decapeptide gonadotrophin-releasing hormone (GnRH) regulates activity of the pituitary-gonadal reproductive axis. Defects of this neuroendocrine axis necessarily result in hypogonadotrophic hypogonadism. In many vertebrate species studied, the main population of GnRH neurones originates extracranially within the olfactory system. In humans, both olfactory and GnRH systems are affected in Kallmann's syndrome--resulting in isolated hypogonadotrophic hypogonadism (IHH) combined with anosmia (loss of sense of smell). Familial IHH is also caused by other genetic conditions, which prevent GnRH from activating luteinizing hormone/follicle-stimulating hormone release from pituitary gonadotrophs. However, many cases of IHH have no defined chromosomal abnormality and, in the absence of pedigree analysis, studying the biological mechanisms controlling migration of GnRH neurones through the olfactory system into the developing central nervous system might reveal additional genetic pathways that play a role in the pathogenesis of IHH. PMID- 11893525 TI - Regulation of cholesterol supply for mineralocorticoid biosynthesis. AB - In addition to intracellular cholesterol synthesis, plasma low- and high-density lipoproteins (LDL and HDL, respectively) are the major potential sources of a cholesterol precursor for steroid synthesis in all steroidogenic tissues. LDL- and HDL-cholesterol are taken up by cells through entirely distinct mechanisms. In the case of aldosterone production in the zona glomerulosa of the adrenal cortex, it has been assumed in the past that LDL is the major supplier of cholesterol. However, recent developments, in particular the discovery of the scavenger receptor class B type I for HDL and the characterization of its properties, have questioned this view. In fact, the nature of the challenging factor (angiotensin II or adrenocorticotropic hormone) appears to determine which pool of cholesterol is preferentially mobilized and which pathway (LDL receptor endocytosis or selective uptake through the HDL receptor) is regulated. PMID- 11893526 TI - Mechanisms in tissue-specific regulation of estrogen biosynthesis in humans. AB - In humans, aromatase P450, which catalyses conversion of C(19)-steroids to estrogens, is expressed in several tissues, including gonads, brain, adipose tissue, skin and placenta, and is encoded by a single-copy gene (CYP19); however, this does not hold true for all species. The human gene is approximately 130 kb and its expression is regulated, in part, by tissue-specific promoters and by alternative splicing mechanisms. Using transgenic mouse technology, it was observed that ovary-, adipose tissue- and placenta-specific expression of human CYP19 is directed by relatively small segments of DNA within 500 bp upstream of each of the tissue-specific first exons. Thus, the use of alternative promoters allows greater versatility in tissue-specific regulation of CYP19 expression. Characterization and identification of transcription factors and crucial cis acting elements within genomic regions that direct tissue-specific expression will contribute to improved understanding of the regulation of CYP19 expression in the tissues that synthesize estrogens under both physiological and pathophysiological conditions. PMID- 11893533 TI - The academic career as a platform for launching a company. PMID- 11893534 TI - Overcoming drug-resistant yeast infections. PMID- 11893527 TI - Neuropeptides as growth factors for normal and cancerous cells. AB - Neuropeptides are molecular messengers that regulate multiple functions in the central nervous system and in the periphery via G-protein-coupled receptors. These signaling peptides have also been identified as potent cellular growth factors for normal cells and they participate in autocrine/paracrine stimulation of tumor cell proliferation and migration. Recent studies on the signaling pathways activated by mitogenic neuropeptides revealed previously unsuspected connections and complexities, including the realization that these receptors not only stimulate the synthesis of conventional second messengers but also induce tyrosine phosphorylation cascades. A major task for the future will be to identify all the contributing molecules, define their functional importance and elucidate the spatiotemporal relationships of this complicated signaling network. As our understanding of the role of neuropeptides in cancer increases, novel possibilities for translational research are emerging for improving the diagnosis and treatment of the disease. PMID- 11893535 TI - Viral Trojan horse for combating tuberculosis. PMID- 11893536 TI - A 'C' change for hepatitis treatment. PMID- 11893539 TI - The way to a powerful business plan. PMID- 11893540 TI - Gene patents: are they socially acceptable monopolies, essential for drug discovery? -- reply. PMID- 11893541 TI - Finding new antibiotics: the power of computational methods. PMID- 11893542 TI - Resecting the neck of the medicine bottle. PMID- 11893543 TI - Metabolic profiling: pathways in drug discovery. PMID- 11893544 TI - Cellular platforms for HTS: three case studies. AB - The field of cell-based screening is expanding rapidly as innovations in target selection and instrumentation increase the number of targets that can be efficiently screened in cellular formats. Cell-based screens can be configured to provide a broad range of data on chemical compound activity, mechanism of action and drugability. However, the decision to pursue a cell-based approach should not be made lightly, as cell-based assays can be challenging to implement in the high throughput screening (HTS) laboratory. In this review, we describe three case studies in which targets were successfully interrogated in cell-based HTS, and highlight the necessary steps to ensure the validity of these screens. PMID- 11893545 TI - Metabolic profiling of cell growth and death in cancer: applications in drug discovery. AB - Metabolic profiling using stable-isotope tracer technology enables the measurement of substrate redistribution within major metabolic pathways in living cells. This technique has demonstrated that transformed human cells exhibit profound metabolic shifts and that some anti-cancer drugs produce their effects by forcing the reversion of these metabolic changes. By revealing tumor-specific metabolic shifts in tumor cells, metabolic profiling enables drug developers to identify the metabolic steps that control cell proliferation, thus aiding the identification of new anti-cancer targets and screening of lead compounds for anti-proliferative metabolic effects. PMID- 11893546 TI - The impact of microwave-assisted organic chemistry on drug discovery. AB - Microwave-assisted organic synthesis (MAOS) is rapidly becoming recognized as a valuable tool for easing some of the bottlenecks in the drug discovery process. This article outlines the basic principles behind the technology and summarizes the areas in which microwave technology has made an impact, to date. PMID- 11893549 TI - Inherited sodium channelopathies: models for acquired arrhythmias? PMID- 11893550 TI - beta(1)-Receptors increase cAMP and induce abnormal Ca(i) cycling in the German shepherd sudden death model. AB - We studied the role of beta-adrenergic receptor subtype signaling to cAMP and calcium in the genesis of catecholamine-dependent arrhythmias in German shepherd dogs that develop lethal arrhythmias at ~5 mo of age. There were three major findings in this study: 1) isoproterenol induces similar increases in cAMP in afflicted and control dogs exclusively through beta(1)-receptors (not beta(2)), 2) cells from afflicted dogs display prolonged relaxation kinetics at long cycle lengths and large frequent spontaneous calcium oscillations (and aftercontractions) with little increase in calcium transient amplitude in response to beta(1)-receptor agonists, and 3) beta(2)-receptor agonists induce a similar marked increases in calcium transient and twitch amplitude, with only rare spontaneous calcium oscillations in afflicted and control cells. These results indicate that catecholamines provide inotropic support to canine cardiomyocytes through distinct beta(1)- and beta(2)-receptor pathways with differing requirements for cAMP. The propensity to develop arrhythmias is not induced by beta(2)-receptors (or a rise in calcium alone), but rather occurs in the context of beta(1)-receptor activation of the cAMP-dependent pathway. PMID- 11893551 TI - Purkinje involvement in arrhythmias after coronary artery reperfusion. AB - Previous studies have indicated that the endocardium may be responsible for a large portion of ventricular tachycardia (VT) seen with reperfusion of ischemic myocardium. To evaluate the role of the Purkinje system in nonreentrant VT arising from the endocardium after reperfusion, the anterior descending coronary artery was occluded for 20 min and then reperfused in 23 dogs after instrumentation of the risk zone with 21 multipolar plunge needles. VT with focal Purkinje origin was defined as a focal endocardial VT with Purkinje potentials recorded before the earliest endocardial myopotential. A total of 19 VTs (mean cycle length 214 +/- 2 ms) were observed with 11 (58%) having focal Purkinje origin. Fifty-eight percent of the VTs degenerated to ventricular fibrillation, with occurrences of two or more independent foci per complex (seen in 7 of 11 compared with 1 of 8 nonsustained VTs). In conclusion, these data show that Purkinje tissue may be important in the genesis of reperfusion VT. PMID- 11893552 TI - Apoptosis in the left ventricle of chronic volume overload causes endocardial endothelial dysfunction in rats. AB - The hypothesis is that chronic increases in left ventricular (LV) load induce oxidative stress and latent matrix metalloproteinase (MMP) is activated, allowing the heart to dilate in the absence of endothelial nitric oxide (NO) and thereby reduce filling pressure. To create volume overload, an arteriovenous (A-V) fistula was placed in male Sprague-Dawley rats. To decrease oxidative stress and apoptosis, 0.08 mg/ml nicotinamide (Nic) was administered in drinking water 2 days before surgery. The rats were divided into the following groups: 1) A-V fistula, 2) A-V fistula + Nic, 3) sham operated, 4) sham + Nic, and 5) control (unoperated); n = 6 rats/group. After 4 wk, hemodynamic parameters were measured in anesthetized rats. The heart was removed and weighed, and LV tissue homogeneates were prepared. A-V fistula caused an increase in heart weight, lung weight, and end-diastolic pressure compared with the sham group. The levels of malondialdehyde (MDA; a marker of oxidative stress) was 6.60 +/- 0.23 ng/mg protein and NO was 6.87 +/- 1.21 nmol/l in the LV of A-V fistula rats by spectrophometry. Nic treatment increased NO to 13.88 +/- 2.5 nmol/l and decreased MDA to 3.54 +/- 0.34 ng/mg protein (P = 0.005). Zymographic levels of MMP-2 were increased, as were protein levels of nitrotyrosine and collagen fragments by Western blot analysis. The inhibition of oxidative stress by Nic decreased nitrotyrosine content and MMP activity. The levels of tissue inhibitor of metalloproteinase-4 mRNA were decreased in A-V fistula rats and increased in A-V fistula rats treated with Nic by Northern blot analysis. TdT-mediated dUTP nick end labeling-positive cells were increased in A-V fistula rats and decreased in fistula rats treated with Nic. Acetylcholine and nitroprusside responses in cardiac rings prepared from the above groups of rats suggest impaired endothelial dependent cardiac relaxation. Treatment with Nic improves cardiac relaxation. The results suggest that an increase in the oxidative stress and generation of nitrotyrosine are, in part, responsible for the activation of metalloproteinase and decreased endocardial endothelial function in chronic LV volume overload. PMID- 11893553 TI - AT(1) and AT(2) receptor expression and blockade after acute ischemia-reperfusion in isolated working rat hearts. AB - We assessed ANG II type 1 (AT(1)) and type 2 (AT(2)) receptor (R) expression and functional recovery after ischemia-reperfusion with or without AT(1)R/AT(2)R blockade in isolated working rat hearts. Groups of six hearts were subjected to global ischemia (30 min) followed by reperfusion (30 min) and exposed to no drug and no ischemia-reperfusion (control), ischemia-reperfusion and no drug, and ischemia-reperfusion with losartan (an AT(1)R antagonist; 1 micromol/l), PD 123319 (an AT(2)R antagonist; 0.3 micromol/l), N(6)-cyclohexyladenosine (CHA, a cardioprotective adenosine A(1) receptor agonist; 0.5 micromol/l as positive control), enalaprilat (an ANG-converting enzyme inhibitor; 1 micromol/l), PD 123319 + losartan, ANG II (1 nmol/l), or ANG II + losartan. Compared with controls, ischemia-reperfusion decreased AT(2)R protein (Western immunoblots) and mRNA (Northern immunoblots, RT-PCR) and impaired functional recovery. PD-123319 increased AT(2)R protein and mRNA and improved functional recovery. Losartan increased AT(1)R mRNA (but not AT(1)R/AT(2)R protein) and impaired recovery. Other groups (except CHA) did not improve recovery. The results suggest that, in isolated working hearts, AT(2)R plays a significant role in ischemia-reperfusion and AT(2)R blockade induces increased AT(2)R protein and cardioprotection. PMID- 11893554 TI - Overexpression of alcohol dehydrogenase exacerbates ethanol-induced contractile defect in cardiac myocytes. AB - Alcoholic cardiomyopathy is characterized by impaired ventricular function although its toxic mechanism is unclear. This study examined the impact of cardiac overexpression of alcohol dehydrogenase (ADH), which oxidizes ethanol into acetaldehyde (ACA), on ethanol-induced cardiac contractile defect. Mechanical and intracellular Ca(2+) properties were evaluated in ventricular myocytes from ADH transgenic and wild-type (FVB) mice. ACA production was assessed by gas chromatography. ADH myocytes exhibited similar mechanical properties but a higher efficiency to convert ACA compared with FVB myocytes. Acute exposure to ethanol depressed cell shortening and intracellular Ca(2+) in the FVB group with maximal inhibitions of 23.3% and 23.4%, respectively. Strikingly, the ethanol-induced depression on cell shortening and intracellular Ca(2+) was significantly augmented in the ADH group, with maximal inhibitions of 43.7% and 40.6%, respectively. Pretreatment with the ADH inhibitor 4 methylpyrazole (4-MP) or the aldehyde dehydrogenase inhibitor cyanamide prevented or augmented the ethanol-induced inhibition, respectively, in the ADH but not the FVB group. The ADH transgene also substantiated the ethanol-induced inhibition of maximal velocity of shortening/relengthening and unmasked an ethanol-induced prolongation of the duration of shortening/relengthening, which was abolished by 4-MP. These data suggest that elevated cardiac ACA exposure due to enhanced ADH expression may play an important role in the development of alcoholic cardiomyopathy. PMID- 11893555 TI - Oxytocin does not directly affect vascular tone in vessels from nonpregnant and pregnant rats. AB - Recent evidence suggests oxytocin (OT) may regulate vascular tone. OT and its receptor (OTR) have been identified in the rat heart and great vessels. Expression of OT and OTR is increased in some tissues during pregnancy. We hypothesized that the OT/OTR system may be a physiological regulator of vascular tone and mediate the decreased vascular resistance noted during pregnancy. Using a wire myograph system, we measured changes in vascular tone in response to OT in small mesenteric arteries, uterine arcuate arteries, and thoracic aorta from nonpregnant and pregnant rats. Additionally, we used reverse transcriptase polymerase chain reaction (RT-PCR) to measure mRNA for OTR in these vascular tissues. Although OTR mRNA was identified by RT-PCR, OT did not elicit a vasodilatory effect in any of the vessels studied. High concentrations of OT (>10(-8) M) caused vasoconstriction that was eliminated by a specific vasopressin V(1a) receptor antagonist. Although it may have an indirect effect in regulation of peripheral resistance, we conclude that OT is unlikely to play a direct role in the physiological regulation of vascular tone. PMID- 11893556 TI - Role of ADP-ribose in 11,12-EET-induced activation of K(Ca) channels in coronary arterial smooth muscle cells. AB - We recently reported that cADP-ribose (cADPR) and ADP-ribose (ADPR) play an important role in the regulation of the Ca(2+)-activated K(+) (K(Ca)) channel activity in coronary arterial smooth muscle cells (CASMCs). The present study determined whether these novel signaling nucleotides participate in 11,12 epoxyeicosatrienoic acid (11,12-EET)-induced activation of the K(Ca) channels in CASMCs. HPLC analysis has shown that 11,12-EET increased the production of ADPR but not the formation of cADPR. The increase in ADPR production was due to activation of NAD glycohydrolase as measured by a conversion rate of NAD into ADPR. The maximal conversion rate of NAD into ADPR in coronary homogenate was increased from 2.5 +/- 0.2 to 3.4 +/- 0.3 nmol*(-1) *mg protein(-1) by 11,12-EET. The regioisomers of 8,9-EET, 11,12-EET, and 14,15-EET also significantly increased ADPR production from NAD. Western blot analysis and immunoprecipitation demonstrated the presence of NAD glycohydrolase, which mediated 11,12-EET activated production of ADPR. In cell-attached patches, 11,12-EET (100 nM) increases K(Ca) channel activity by 5.6-fold. The NAD glycohydrolase inhibitor cibacron blue 3GA (3GA, 100 microM) significantly attenuated 11,12-EET-induced increase in the K(Ca) channel activity in CASMCs. However, 3GA had no effect on the K(Ca) channels activity in inside-out patches. 11,12-EET produced a concentration-dependent relaxation of precontracted coronary arteries. This 11,12 EET-induced vasodilation was substantially attenuated by 3GA (30 microM) with maximal inhibition of 57%. These results indicate that 11,12-EET stimulates the production of ADPR and that intracellular ADPR is an important signaling molecule mediating 11,12-EET-induced activation of the K(Ca) channels in CASMCs and consequently results in vasodilation of coronary artery. PMID- 11893557 TI - Depressed transient outward potassium current density in catecholamine-depleted rat ventricular myocytes. AB - The effect of catecholamine depletion (induced by prior treatment with reserpine) was studied in Wistar rat ventricular myocytes using whole cell voltage-clamp methods. Two calcium-independent outward currents, the transient outward potassium current (I(to)) and the sustained outward potassium current (I(sus)), were measured. Reserpine treatment decreased tissue norepinephrine content by 97%. Action potential duration in the isolated perfused heart was significantly increased in reserpine-treated hearts. In isolated ventricular myocytes, I(to) density was decreased by 49% in reserpine-treated rats. This treatment had no effect on I(sus). The I(to) steady-state inactivation-voltage relationship and recovery from inactivation remained unchanged, whereas the conductance-voltage activation curve for reserpine-treated rats was significantly shifted (6.7 mV) toward negative potentials. The incubation of myocytes with 10 microM norepinephrine for 7-10 h restored I(to), an effect that was abolished by the presence of actinomycin D. Norepinephrine (0.5 microM) had no effect on I(to). However, in the presence of both 0.5 microM norepinephrine and neuropeptide Y (0.1 microM), I(to) density was restored to its control value. These results suggest that the sympathetic nervous system is involved in I(to) regulation. Sympathetic norepinephrine depletion decreased the number of functional channels via an effect on the alpha-adrenergic cascade and norepinephrine is able to restore expression of I(to) channels. PMID- 11893558 TI - Intramyocardial pressure measurements in the stage 18 embryonic chick heart. AB - Intramyocardial pressure (IMP) and ventricular pressure (VP) were measured in the trabeculating heart of the stage 18 chick embryo (3 days of incubation). Pressure was measured at several locations across the ventricle using a fluid-filled servo null system. Maximum systolic and minimum diastolic IMP tended to be greater in the dorsal wall than in the ventral wall, but transmural distributions of peak active (maximum minus minimum) IMP were similar in both walls. Peak active IMP near midwall was similar to peak active VP, but peak active IMP in the subepicardial and subendocardial layers was four to five times larger. These results suggest that the passive stiffness of the dorsal wall is greater than that of the ventral wall and that during contraction the inner and outer layers of both walls generate more contractile force and/or become less permeable to flow than the middle part of the wall. Measured pressures likely correspond to regional variations in wall stress that may influence morphogenesis and function in the embryonic heart. PMID- 11893559 TI - Cellular glutathione peroxidase deficiency and endothelial dysfunction. AB - Cellular glutathione peroxidase (GPx-1) is the most abundant intracellular isoform of the GPx antioxidant enzyme family. In this study, we hypothesized that GPx-1 deficiency directly induces an increase in vascular oxidant stress, with resulting endothelial dysfunction. We studied vascular function in a murine model of homozygous deficiency of GPx-1 (GPx-1(-/-)). Mesenteric arterioles of GPx-1(-/ ) mice demonstrated paradoxical vasoconstriction to beta-methacholine and bradykinin, whereas wild-type (WT) mice showed dose-dependent vasodilation in response to both agonists. One week of treatment of GPx-1(-/-) mice with L-2 oxothiazolidine-4-carboxylic acid (OTC), which increases intracellular thiol pools, resulted in restoration of normal vascular reactivity in the mesenteric bed of GPx-1(-/-) mice. We observed an increase of the isoprostane iPF(2alpha) III, a marker of oxidant stress, in the plasma and aortas of GPx-1(-/-) mice compared with WT mice, which returned toward normal after OTC treatment. Aortic sections from GPx-1(-/-) mice showed increased binding of an anti-3-nitrotyrosine antibody in the absence of frank vascular lesions. These findings demonstrate that homozygous deficiency of GPx-1 leads to impaired endothelium-dependent vasodilator function presumably due to a decrease in bioavailable nitric oxide and to increased vascular oxidant stress. These vascular abnormalities can be attenuated by increasing bioavailable intracellular thiol pools. PMID- 11893560 TI - Influence of vascular dimension on gender difference in flow-dependent dilatation of peripheral conduit arteries. AB - To assess the influence of initial diameter on the gender difference in flow dependent dilatation (FDD) of the conduit artery, we measured radial artery internal diameter (echotracking), flow (Doppler) and total blood viscosity in 24 healthy (25 +/- 0.8 yr) men and women during reactive hyperemia (RH) and during a gradual hand skin heating (SH). At baseline, mean diameter (men, 2.76 +/- 0.09 vs. women, 2.32 +/- 0.07 mm, P < 0.05), flow (men, 21 +/- 4 vs. women, 10 +/- 1 ml/min, P < 0.05), and blood viscosity (men, 4.13 +/- 0.07 vs. women, 3.92 +/- 0.13 cP, P < 0.05) were higher in men but mean shear stress (MSS) was not different between groups. During RH, the percent increase in diameter was lower in men (men, 9 +/- 1 vs. women, 13 +/- 1%, P < 0.05). This difference was suppressed after correction for baseline diameter. During SH, the increase in diameter with flow was higher in women (P < 0.01). However, the increase in MSS was higher in women because of their smaller diameter at each level of flow (P < 0.01) and there was no difference between groups for the increase in diameter at each level of MSS. These results demonstrate in a direct manner that initial diameter influences the magnitude of FDD of conduit arteries in humans by modifying the value of the arterial wall shear stress at each level of flow and support the interest of the heating method in presence of heterogeneous groups. PMID- 11893561 TI - Biphasic effects of cell volume on excitation-contraction coupling in rabbit ventricular myocytes. AB - We studied the effects of osmotic swelling on the components of excitation contraction coupling in ventricular myocytes. Myocyte volume rapidly increased 30% in hyposmotic (0.6T) solution and was constant thereafter. Cell shortening transiently increased 31% after 4 min in 0.6T but then decreased to 68% of control after 20 min. In parallel, the L-type Ca(2+) current (I(Ca-L)) transiently increased 10% and then declined to 70% of control. Similar biphasic effects on shortening were observed under current clamp. In contrast, action potential duration was unchanged at 4 min but decreased to 72% of control after 20 min. Ca(2+) transients were measured with fura 2-AM. The emission ratio with excitation at 340 and 380 nm (f(340)/f(380)) decreased by 12% after 3 min in 0.6T, whereas shortening and I(Ca-L) increased at the same time. After 8 min, shortening, I(Ca-L), and the f(340)/f(380) ratio decreased 28, 25, and 59%, respectively. The results suggest that osmotic swelling causes biphasic changes in I(Ca-L) that contribute to its biphasic effects on contraction. In addition, swelling initially appears to reduce the Ca(2+) transient initiated by a given I(Ca-L), and later, both I(Ca-L) and the Ca(2+) transient are inhibited. PMID- 11893562 TI - Integration of cornea and cardiorespiratory afferents in the nucleus of the solitary tract of the rat. AB - We determined the activity of neurons within the nucleus of the solitary tract (NTS) after stimulation of the cornea and assessed whether this input affected the processing of baroreceptor and peripheral chemoreceptor inputs. In an in situ, unanesthetized decerebrate working heart-brain stem preparation of the rat, noxious mechanical or electrical stimulation was applied to the cornea, and extracellular single unit recordings were made from NTS neurons. Cornea nociceptor stimulation evoked bradycardia and an increase in the cycle length of the phrenic nerve discharge. Of 90 NTS neurons with ongoing activity, corneal stimulation excited 51 and depressed 39. There was a high degree of convergence to these NTS neurons from either baroreceptors or chemoreceptors. The excitatory synaptic response in 12 of 19 baroreceptive and 10 of 15 chemoreceptive neurons was attenuated significantly during concomitant electrical stimulation of the cornea. This inhibition was GABA(A) receptor mediated, being blocked by pressure ejection of bicuculline. Thus the NTS integrates information from corneal receptors, some of which converges onto neurons mediating reflexes from baroreceptors and chemoreceptors to inhibit these inputs. PMID- 11893563 TI - TNF-alpha and myocardial matrix metalloproteinases in heart failure: relationship to LV remodeling. AB - The cytokine tumor necrosis factor (TNF)-alpha has been causally linked to left ventricular (LV) remodeling, but the molecular basis for this effect is unknown. Matrix metalloproteinases (MMPs) have been implicated in cardiac remodeling and can be regulated by TNF-alpha. This study tested the central hypothesis that administration of a TNF-alpha blocking protein would prevent the induction of MMPs and alter the course of myocardial remodeling in developing LV failure. Adult dogs were randomly assigned to the following groups: 1) chronic pacing (250 beats/min, 28 days, n = 12), 2) chronic pacing with concomitant administration of a TNF-alpha blocking protein (TNF block) using a soluble p75 TNF receptor fusion protein (TNFR:Fc; administered at 0.5 mg/kg twice a week subcutaneously, n = 7), and 3) normal controls (n = 10). LV end-diastolic volume increased from control with chronic pacing (83 +/- 12 vs. 118 +/- 10 ml, P < 0.05) and was reduced with TNF block (97 +/- 9 ml, P < 0.05). MMP zymographic levels (92 kDa, pixels) increased from control with chronic pacing (36,848 +/- 9,593 vs. 87,247 +/- 12,912, P < 0.05) and was normalized by TNF block. Myocardial MMP-9 and MMP-13 levels by immunoblot increased with chronic pacing relative to controls (130 +/- 10% and 118 +/- 6%, P < 0.05) and was normalized by TNF block. These results provide evidence to suggest that TNF-alpha contributes to the myocardial remodeling process in evolving heart failure through the local induction of specific MMPs. PMID- 11893564 TI - Role of myocardium and endothelium in coronary vascular smooth muscle responses to hypoxia. AB - Hypoxia triggers a mechanism that induces vasodilation in the whole heart but not necessarily in isolated coronary arteries. We therefore studied the role of cardiomyocytes (CM), smooth muscle cells (SMC), and endothelial cells (EC) in coronary responses to hypoxia (PO(2) of 5-10 mmHg). In an attempt to determine the factor(s) released in response to hypoxia, we inhibited the contribution of adenosine, ATP-sensitive K(+) channels, prostaglandins, and nitric oxide. Isolated rat septal artery segments without (-T) and with a layer of cardiac tissue (+T) were mounted in a double wire myograph, and constriction was induced. Hypoxia induced a decrease in isometric force of 21% and 61% in -T and +T segments, respectively (P < 0.05). EC removal increased the relaxation to hypoxia in -T segments to 33% but had the same effect in +T segments (61%). Only one of the inhibitors, the adenosine antagonist in +T segments, partially affected the relaxation due to hypoxia. The role of adenosine is thus limited and other mechanisms have to contribute. We conclude that hypoxia induces a relaxation of SMC that is augmented by the presence of CM and blunted by the endothelium. A single mediator does not induce those effects. PMID- 11893565 TI - Role of FKBP12.6 in cADPR-induced activation of reconstituted ryanodine receptors from arterial smooth muscle. AB - cADP ribose (cADPR) serves as second messenger to activate the ryanodine receptors (RyRs) of the sarcoplasmic reticulum (SR) and mobilize intracellular Ca(2+) in vascular smooth muscle cells. However, the mechanisms mediating the effect of cADPR remain unknown. The present study was designed to determine whether FK-506 binding protein 12.6 (FKBP12.6), an accessory protein of the RyRs, plays a role in cADPR-induced activation of the RyRs. A 12.6-kDa protein was detected in bovine coronary arterial smooth muscle (BCASM) and cultured CASM cells by being immunoblotted with an antibody against FKBP12, which also reacted with FKBP12.6. With the use of planar lipid bilayer clamping techniques, FK-506 (0.01-10 microM) significantly increased the open probability (NP(O)) of reconstituted RyR/Ca(2+) release channels from the SR of CASM. This FK-506 induced activation of RyR/Ca(2+) release channels was abolished by pretreatment with anti-FKBP12 antibody. The RyRs activator cADPR (0.1-10 microM) markedly increased the activity of RyR/Ca(2+) release channels. In the presence of FK-506, cADPR did not further increase the NP(O) of RyR/Ca(2+) release channels. Addition of anti-FKBP12 antibody also completely blocked cADPR-induced activation of these channels, and removal of FKBP12.6 by preincubation with FK-506 and subsequent gradient centrifugation abolished cADPR-induced increase in the NP(O) of RyR/Ca(2+) release channels. We conclude that FKBP12.6 plays a critical role in mediating cADPR-induced activation of RyR/Ca(2+) release channels from the SR of BCASM. PMID- 11893566 TI - Phase I and phase II of short-term mechanical restitution in perfused rat left ventricles. AB - We examined the contributions of the Ca(2+) channels of the sarcolemma and of the sarcoplasmic reticulum to electromechanical restitution. Extrasystoles (F(1)) were interpolated 40-600 ms following a steady-state beat (F(0)) in perfused rat ventricles paced at 2 or 3 Hz. Plots of F(1)/F(0) versus the extrasystolic interval consisted of phase I, which occurred before relaxation of the steady state beat, and phase II, which occurred later. Phase I exhibited a period of enhanced left ventricular pressure development that coincided with action potential prolongation. Phase I was eliminated by -BAY K 8644 (100 nM) and FPL 64176 (150 nM), augmented by 3 microM thapsigargin plus 200 nM ryanodine and unaffected by KN-93 and KB-R7943. Phase II was accelerated by the Ca(2+) channel agonists and by isoproterenol but was eliminated by thapsigargin plus ryanodine. The results suggest that phase I of electromechanical restitution is caused by a transient L-type Ca(2+) current facilitation, whereas phase II represents the recovery of the ability of the sarcoplasmic reticulum to release Ca(2+). PMID- 11893567 TI - Mechanical alternans and restitution in failing SHHF rat left ventricles. AB - We examined mechanical alternans and electromechanical restitution in normal and failing rat hearts. Alternans occurred at 5 Hz in failing versus 9 Hz in control hearts and was reversed by 300 nM isoproterenol, 6 mM extracellular Ca(2+), 300 nM -BAY K 8644, or 50 nM ryanodine. Restitution curves comprised phase I, which was completed before relaxation of the steady-state beat, and phase II, which occurred later. Phase I action potential area and developed pressure ratios were significantly reduced in the failing versus control hearts. Phase II was a monoexponential increase in relative developed pressure as the extrasystolic interval was increased. The plateau of phase II was significantly elevated in failing hearts. Thapsigargin (3 microM) plus ryanodine (200 nM) potentiated phase I to a significantly greater extent in control versus failing hearts and abolished phase II in both groups. The results suggest that both regulation of Ca(2+) influx across the sarcolemma and Ca(2+) release by the sarcoplasmic reticulum may contribute to altered excitation-contraction coupling in the failing spontaneously hypertensive heart failure prone rat heart. PMID- 11893568 TI - Ventricular remodeling and diastolic myocardial dysfunction in rats submitted to protein-calorie malnutrition. AB - The effects of protein-calorie malnutrition (PCM) on heart structure and function are not completely understood. We studied heart morphometric, functional, and biochemical characteristics in undernourished young Wistar rats. They were submitted to PCM from birth (undernourished group, UG). After 10 wk, left ventricle function was studied using a Langendorff preparation. The results were compared with age-matched rats fed ad libitum (control group, CG). The UG rats achieved 47% of the body weight and 44% of the left ventricular weight (LVW) of the CG. LVW-to-ventricular volume ratio was smaller and myocardial hydroxyproline concentration was higher in the UG. Left ventricular systolic function was not affected by the PCM protocol. The myocardial stiffness constant was greater in the UG, whereas the end-diastolic pressure-volume relationship was not altered. In conclusion, the heart is not spared from the adverse effects of PCM. There is a geometric alteration in the left ventricle with preserved ventricular compliance despite the increased passive myocardial stiffness. The systolic function is preserved. PMID- 11893569 TI - Increased coronary perfusion augments cardiac contractility in the rat through stretch-activated ion channels. AB - The role of stretch-activated ion channels (SACs) in coronary perfusion-induced increase in cardiac contractility was investigated in isolated isometrically contracting perfused papillary muscles from Wistar rats. A brief increase in perfusion pressure (3-4 s, perfusion pulse, n = 7), 10 repetitive perfusion pulses (n = 4), or a sustained increase in perfusion pressure (150-200 s, perfusion step, n = 7) increase developed force by 2.7 +/- 1.1, 7.7 +/- 2.2, and 8.3 +/- 2.5 mN/mm(2) (means +/- SE, P < 0.05), respectively. The increase in developed force after a perfusion pulse is transient, whereas developed force during a perfusion step remains increased by 5.1 +/- 2.5 mN/mm(2) (P < 0.05) in the steady state. Inhibition of SACs by addition of gadolinium (10 micromol/l) or streptomycin (40 and 100 micromol/l) blunts the perfusion-induced increase in developed force. Incubation with 100 micromol/l N(omega)-nitro-L-arginine [nitric oxide (NO) synthase inhibition], 10 micromol/l sodium nitroprusside (NO donation) and 0.1 micromol/l verapamil (L-type Ca(2+) channel blockade) are without effect on the perfusion-induced increase of developed force. We conclude that brief, repetitive, or sustained increases in coronary perfusion augment cardiac contractility through activation of stretch-activated ion channels, whereas endothelial NO release and L-type Ca(2+) channels are not involved. PMID- 11893570 TI - Acetaminophen and low-flow myocardial ischemia: efficacy and antioxidant mechanisms. AB - In the current study, the cardioprotective efficacy of 0.35 mmol/l acetaminophen administered 10 min after the onset of a 20-min period of global, low-flow myocardial ischemia was investigated. Matched control hearts were administered an equal volume of Krebs-Henseleit physiological buffer solution (vehicle). In separate groups of hearts, the concentration-dependent, negative inotropic properties of hydrogen peroxide and the ability of acetaminophen to attenuate these actions, as well as the effects of acetaminophen on ischemia-reperfusion mediated protein oxidation, were studied. Acetaminophen-treated hearts regained a significantly greater fraction of baseline, preischemia control function during reperfusion than vehicle-treated hearts. For example, contractility [rate of maximal developed pressure in the left ventricle (+/-dP/dt(max))] after 10 min of reperfusion was 109 +/- 24 and 42 +/- 9 mmHg/s (P < 0.05), respectively, in the two groups. The corresponding pressure-rate products were 1,840 +/- 434 vs. 588 +/- 169 mmHg*beats*min(-1) (P < 0.05). Acetaminophen attenuated peroxynitrite mediated chemiluminescence in the early minutes of reperfusion (e.g., at 6 min, corresponding values for peak light production were approximately 8 x 10(6) counts/min for vehicle vs. <4 x 10(6) counts/min for acetaminophen, P < 0.05) and the negative inotropic effects of exogenously administered hydrogen peroxide (e.g., at 0.4 mmol/l hydrogen peroxide, pressure-rate products were approximately 1.0 x 10(4) and 3.8 x 10(3) mmHg*beats*min(-1) in acetaminophen- and vehicle treated hearts, respectively, P < 0.05). Ischemia-mediated protein oxidation was reduced by acetaminophen. The ability of acetaminophen to attenuate the damaging effects of peroxynitrite and hydrogen peroxide and to limit protein oxidation suggest antioxidant mechanisms are responsible for its cardioprotective properties during postischemia-reperfusion. PMID- 11893571 TI - Indexes of diastolic RV function: load dependence and changes after chronic RV pressure overload in lambs. AB - Diastolic function is a major determinant of ventricular performance, especially when loading conditions are altered. We evaluated biventricular diastolic function in lambs and studied possible load dependence of diastolic parameters [minimum first derivative of pressure vs. time (dP/dt(min)) and time constant of isovolumic relaxation (tau)] in normal (n = 5) and chronic right ventricular (RV) pressure-overloaded (n = 5) hearts by using an adjustable band on the pulmonary artery (PAB). Pressure-volume relations were measured during preload reduction to obtain the end-diastolic pressure-volume relationship (EDPVR). In normal lambs, absolute dP/dt(min) and tau were lower in the RV than in the left ventricle whereas the chamber stiffness constant (b) was roughly the same. After PAB, RV tau and dP/dt(min) were significantly higher compared with control. The RV EDPVR indicated impaired diastolic function. During acute pressure reduction, both dP/dt(min) and tau showed a relationship with end-systolic pressure. These relationships could explain the increased dP/dt(min) but not the increased tau value after banding. Therefore, the increased tau after banding reflects intrinsic myocardial changes. We conclude that after chronic RV pressure overload, RV early relaxation is prolonged and diastolic stiffness is increased, both indicative of impaired diastolic function. PMID- 11893572 TI - Effects of gestational age on myocardial blood flow and coronary flow reserve in pressure-loaded ovine fetal hearts. AB - To test the hypothesis that coronary flow and coronary flow reserve are developmentally regulated, we used fluorescent microspheres to investigate the effects of acute (6 h) pulmonary artery banding (PAB) on baseline and adenosine enhanced right (RV) and left ventricular (LV) blood flow in two groups of twin ovine fetuses (100 and 128 days of gestation, term 145 days, n = 6 fetuses/group). Within each group, one fetus underwent PAB to constrict the main pulmonary artery diameter by 50%, and the other twin served as a nonbanded control. Physiological measurements were made 6 h after the surgery was completed; tissues were then harvested for analysis of selected genes that may be involved in the early phase of coronary vascular remodeling. Within each age group, arterial blood gas values, heart rate, and mean arterial blood pressure were similar between control and PAB fetuses. Baseline endocardial blood flow in both ventricles was greater in 100 than 128-day fetuses (RV: 341 +/- 20 vs. 230 +/- 17 ml*min(-1)*100 g(-1); LV: 258 +/- 18 vs. 172 +/- 23 ml*min(-1)*100 g(-1), both P < 0.05). In both age groups, RV and LV endocardial blood flows increased significantly in control animals during adenosine infusion and were greater in PAB compared with control fetuses. After PAB, adenosine further increased RV blood flow in 128-day fetuses (from 416 +/- 30 to 598 +/- 33 ml*min(-1)*g(-1), P < 0.05) but did not enhance blood flow in 100-day animals (490 +/- 59 to 545 +/- 42 ml*min(-1)*100 g(-1), P > 0.2). RV vascular endothelial growth factor and Flk 1 mRNA levels were increased relative to controls (P < 0.05) in 128 but not 100 day PAB fetuses. We conclude that in the ovine fetus, developmentally related differences exist in 1) baseline myocardial blood flows, 2) the adaptive response of myocardial blood flow to acute systolic pressure load, and 3) the responses of selected genes involved in vasculogenesis to increased load in the fetal myocardium. PMID- 11893573 TI - Ischemic cardiomyopathy in pigs with two-vessel occlusion and viable, chronically dysfunctional myocardium. AB - A chronic left anterior descending coronary artery (LAD) stenosis leads to the development of hibernating myocardium with severe regional hypokinesis but normal global ventricular function after 3 mo. We hypothesized that two-vessel occlusion would accelerate the progression to hibernating myocardium and lead to global left ventricular (LV) dysfunction and heart failure. Pigs were instrumented with a fixed 1.5-mm constrictor on the proximal LAD and circumflex arteries. After 2 mo, there were no overt signs of right-heart failure and triphenyl tetrazolium chloride infarction was trivial (1.4 +/- 0.1% of the LV). Compared with shams, regional function [myocardial systolic excursion (DeltaWT); 2.1 +/- 0.3 vs. 4.6 +/- 0.4 mm, P < 0.05] and resting perfusion (0.90 +/- 0.13 vs. 1.32 +/- 0.09 ml small middle dot min(-1) small middle dot g(-1), P < 0.05) were reduced, consistent with hibernating myocardium. Pulmonary systolic (45.9 +/- 3.3 vs. 36.5 +/- 2.2 mmHg, P < 0.05) and wedge pressures (19.1 +/- 1.6 vs. 11.2 +/- 0.9 mmHg, P < 0.05) were increased with global ventricular dysfunction (ejection fraction 43 +/- 2 vs. 50 +/- 2%, P < 0.05). Early LV remodeling was present with increased cavity size and mass. Reductions in sarcoplasmic reticulum Ca(2+)-ATPase and phospholamban were confined to the dysfunctional LAD region with no change in calsequestrin. Thus combined stenoses of the LAD and circumflex arteries accelerate the development of hibernating myocardium and result in compensated heart failure. PMID- 11893575 TI - Enhancement of reperfusion injury by elevation of microvascular pressures. AB - Elevated venous pressure can be associated with severe tissue injury. Few links, however, between venous hypertension and tissue damage have been established. We examined here the effects of micropressure elevation on the outcome of venular occlusion/reperfusion in the mesenteric microvasculature of male Wistar rats. One hour of venular occlusion (diameter approximately 50 microm) by micropipette occlusion followed by reperfusion were carried out with sham surgery without occlusion as control. Leukocyte rolling, adhesion, and migration, oxygen radicals detected by dichlorofluorescein (DCF), and parenchymal cell death detected by propidium iodide (PI) were recorded simultaneously in the same vessel at a location upstream of the occlusion site with elevated micropressure and at a downstream location with low micropressure. The number of rolling, adhering, and migrating leukocytes increased on the upstream side of the occlusion to a higher level than downstream of the occlusion site. During occlusion, DCF intensity on the venular endothelium was greater on the upstream side than in the downstream side, but there were no differences during reperfusion. The number of PI-positive cells adjacent to the venules increased significantly compared with controls, and it remained greater on the upstream higher-pressure side than the downstream side. Leukocyte adhesion and transvascular migration in postcapillary venules as well as parenchymal cell death could be significantly reduced by the hydroxyl radical scavenger dimethylthiourea. Microhemorrhages of blood cells into the mesentery interstitium were observed only on the upstream side of the occlusion. These results indicate that an elevation of the venular blood pressure during occlusion/reperfusion exacerbates the inflammatory cascade and tissue injury. Venous occlusion may constitute an important mechanism for tissue injury. PMID- 11893574 TI - Preconditioning blocks cardiocyte apoptosis: role of K(ATP) channels and PKC epsilon. AB - The aims of this study were to determine whether preconditioning blocks cardiocyte apoptosis and to determine the role of mitochondrial ATP-sensitive K(+) (K(ATP)) channels and the protein kinase C epsilon-isoform (PKC-epsilon) in this effect. Ventricular myocytes from 10-day-old chick embryos were used. In the control series, 10 h of simulated ischemia followed by 12 h of reoxygenation resulted in 42 +/- 3% apoptosis (n = 8). These results were consistent with DNA laddering and TdT-mediated dUTP nick-end labeling (TUNEL) assay. Preconditioning, elicited with three cycles of 1 min of ischemia separated by 5 min of reoxygenation before subjection to prolonged simulated ischemia, markedly attenuated the apoptotic process (28 +/- 4%, n = 8). The selective mitochondrial K(ATP) channel opener diazoxide (400 micromol/l), given before ischemia, mimicked preconditioning effects to prevent apoptosis (22 +/- 4%, n = 6). Pretreatment with 5-hydroxydecanoate (100 micromol/l), a selective mitochondrial K(ATP) channel blocker, abolished preconditioning (42 +/- 2%, n = 6). In addition, the effects of preconditioning and diazoxide were blocked with the specific PKC inhibitors Go-6976 (0.1 micromol/l) or chelerythrine (4 micromol/l), given at simulated ischemia and reoxygenation. Furthermore, preconditioning and diazoxide selectively activated PKC-epsilon in the particulate fraction before simulated ischemia without effect on the total fraction, cytosolic fraction, and PKC delta isoform. The specific PKC activator phorbol 12-myristate 13-acetate (0.2 micromol/l), added during simulated ischemia and reoxygenation, mimicked preconditioning to block apoptosis. Opening mitochondrial K(ATP) channels blocks cardiocyte apoptosis via activating PKC-epsilon in cultured ventricular myocytes. Through this signal transduction, preconditioning blocks apoptosis and preserves cardiac function in ischemia-reperfusion. PMID- 11893577 TI - N-methyl-D-aspartate-induced vasodilation is mediated by endothelium-independent nitric oxide release in piglets. AB - N-methyl-D-aspartate (NMDA) elicits pial arteriolar dilation that has been associated with neuronal nitric oxide (NO) production. However, endothelial factors or glial P-450 epoxygenase products may play a role. We tested whether NMDA-induced pial vasodilation 1) primarily involves NO diffusion from the parenchyma to the surface arterioles, 2) involves intact endothelial function, and 3) involves a miconazole-sensitive component. Arteriolar diameters were determined using closed cranial window-intravital microscopy in anesthetized piglets. NMDA (10-100 microM) elicited virtually identical dose-dependent dilations in paired arterioles (r = 0.94, n = 15). However, NMDA- but not bradykinin (BK)-induced dilations of arteriolar sections over large veins were reduced by 31 +/- 1% (means +/- SE, P < 0.05, n = 4) compared with adjacent sections on the cortical surface. Also, 100 microM NMDA increased cerebrospinal fluid levels of NO metabolites from 3.7 +/- 1.0 to 5.3 +/- 1.2 microM (P < 0.05, n = 6). Endothelial stunning by intracarotid injection of phorbol 12,13 dibutyrate did not affect NMDA-induced vasodilation but attenuated vascular responses to hypercapnia and BK by approximately 70% (n = 7). Finally, miconazole (n = 6, 20 microM) pretreatment and coapplication with NMDA did not alter vascular responses to NMDA. In conclusion, NMDA appears to dilate pial arterioles exclusively through release and diffusion of NO from neurons to the pial surface in piglets. PMID- 11893576 TI - H(2)O(2) opens mitochondrial K(ATP) channels and inhibits GABA receptors via protein kinase C-epsilon in cardiomyocytes. AB - Oxygen radicals and protein kinase C (PKC) mediate ischemic preconditioning. Using a cultured chick embryonic cardiomyocyte model of hypoxia and reoxygenation, we found that the oxygen radicals generated by ischemic preconditioning were H(2)O(2). Like preconditioning, H(2)O(2) selectively activated the epsilon-isoform of PKC in the particulate compartment and increased cell viability after 1 h of hypoxia and 3 h of reoxygenation. The glutathione peroxidase ebselen (converting H(2)O(2) to H(2)O) and the superoxide dismutase inhibitor diethyldithiocarbamic acid abolished the increased H(2)O(2) and the protection of preconditioning. PKC activation with phorbol 12-myristate 13 acetate increased cell survival; the protection of preconditioning was blocked by epsilonV(1-2), a selective PKC-epsilon antagonist. Similar to preconditioning, the protection of PKC activation was abolished by mitochondrial K(ATP) channel blockade with 5-hydroxydecanoate or by GABA receptor stimulation with midazolam or diazepam. In addition, PKC, mitochondrial ATP-sensitive K(+) (K(ATP)) channels, and GABA receptors had no effects on H(2)O(2) generated by ischemic preconditioning before prolonged hypoxia and reoxygenation. We conclude that H(2)O(2) opens mitochondrial K(ATP) channels and inhibits GABA receptors via activating PKC-epsilon. Through this signal transduction, preconditioning protects ischemic cardiomyocytes. PMID- 11893578 TI - Actin cytoskeletal modulation of pressure-induced depolarization and Ca(2+) influx in cerebral arteries. AB - The objective of this study was to examine the role of the actin cytoskeleton in the development of pressure-induced membrane depolarization and Ca(2+) influx underlying myogenic constriction in cerebral arteries. Elevating intraluminal pressure from 10 to 60 mmHg induced membrane depolarization, increased intracellular cytosolic Ca(2+) concentration ([Ca(2+)](i)) and elicited myogenic constriction in both intact and denuded rat posterior cerebral arteries. Pretreatment with cytochalasin D (5 microM) or latrunculin A (3 microM) abolished constriction but enhanced the [Ca(2+)](i) response; similarly, acute application of cytochalasin D to vessels with tone, or in the presence of 60 mM K(+), elicited relaxation accompanied by an increase in [Ca(2+)](i). The effects of cytochalasin D were inhibited by nifedipine (3 microM), demonstrating that actin cytoskeletal disruption augments Ca(2+) influx through voltage-sensitive L-type Ca(2+) channels. Finally, pressure-induced depolarization was enhanced in the presence of cytochalasin D, further substantiating a role for the actin cytoskeleton in the modulation of ion channel function. Together, these results implicate vascular smooth muscle actin cytoskeletal dynamics in the control of cerebral artery diameter through their influence on membrane potential as well as via a direct effect on L-type Ca(2+) channels. PMID- 11893579 TI - Attenuation of neutrophil-mediated myocardial ischemia-reperfusion injury by a calpain inhibitor. AB - Calpains are ubiquitous neutral cysteine proteases. Although their physiological role has yet to be clarified, calpains seem to be involved in the expression of cell adhesion molecules. Therefore, we hypothesized that a selective calpain inhibitor could attenuate polymorphonuclear (PMN) leukocyte-induced myocardial ischemia-reperfusion (I/R) injury. We examined the effects of the calpain inhibitor Z-Leu-Leu-CHO in isolated ischemic (20 min) and reperfused (45 min) rat hearts perfused with PMNs. Z-Leu-Leu-CHO (10 and 20 microM, respectively) significantly improved left ventricular developed pressure (LVDP) (P < 0.01) and the maximal rate of development of LVDP (P < 0.01) compared with I/R hearts perfused without Z-Leu-Leu-CHO. In addition, Z-Leu-Leu-CHO significantly reduced PMN adherence to the vascular endothelium and subsequent infiltration into the postischemic myocardium (P < 0.01). Moreover, Z-Leu-Leu-CHO significantly inhibited expression of P-selectin on the rat coronary microvascular endothelium (P < 0.01). These results provide evidence that Z-Leu-Leu-CHO significantly attenuates PMN-mediated I/R injury in the isolated perfused rat heart to a significant extent via downregulation of P-selectin expression. PMID- 11893580 TI - Manipulation of chloride flux affects histamine-induced contraction in rabbit basilar artery. AB - Cl(-) efflux induces depolarization and contraction of smooth muscle cells. This study was undertaken to explore the role of Cl(-) flux in histamine-induced contraction in the rabbit basilar artery. Male New Zealand White rabbits (n = 16) weighing 1.8-2.5 kg were euthanized by an overdose of pentobarbital sodium. The basilar arteries were removed for isometric tension recording. Histamine produced a concentration-dependent contraction that was attenuated by the H(1) receptor antagonist chlorpheniramine (10(-8) M) but not by the H(2) receptor antagonist cimetidine (3 x 10(-6) M) in normal Cl(-) Krebs-Henseleit bicarbonate solution (123 mM Cl(-)). The histamine-induced contraction was reduced by the following manipulations: 1) inhibition of Na(+)-K(+)-2Cl(-) cotransporter with bumetanide (3 x 10(-5) and 10(-4) M), 2) bicarbonate-free HEPES solution to disable Cl( )/HCO exchanger, and 3) blockade of Cl(-) channels with the use of niflumic acid, 5-nitro-2-(3-phenylpropylamino) benzoic acid, and indoleacetic acid 94 R-(+) methylindazone. In addition, substitution of extracellular Cl(-) (10 mM) with methanesulfonate acid (113 mM) transiently enhanced histamine-induced contraction. Manipulation of Cl(-) flux affects histamine-induced contraction in the rabbit basilar artery. PMID- 11893581 TI - Time-dependent transients in an ionically based mathematical model of the canine atrial action potential. AB - Ionically based cardiac action potential (AP) models are based on equations with singular Jacobians and display time-dependent AP and ionic changes (transients), which may be due to this mathematical limitation. The present study evaluated transients during long-term simulated activity in a mathematical model of the canine atrial AP. Stimulus current assignment to a specific ionic species contributed to stability. Ionic concentrations were least disturbed with the K(+) stimulus current. All parameters stabilized within 6-7 h. Inward rectifier, Na(+)/Ca(2+) exchanger, L-type Ca(2+), and Na(+)-Cl(-) cotransporter currents made the greatest contributions to stabilization of intracellular [K(+)], [Na(+)], [Ca(2+)], and [Cl(-)], respectively. Time-dependent AP shortening was largely due to the outward shift of Na(+)/Ca(2+) exchange related to intracellular Na(+) (Na) accumulation. AP duration (APD) reached a steady state after approximately 40 min. AP transients also occurred in canine atrial preparations, with the APD decreasing by approximately 10 ms over 35 min, compared with approximately 27 ms in the model. We conclude that model APD and ionic transients stabilize with the appropriate stimulus current assignment and that the mathematical limitation of equation singularity does not preclude meaningful long-term simulations. The model agrees qualitatively with experimental observations, but quantitative discrepancies highlight limitations of long-term model simulations. PMID- 11893582 TI - Reversibility of electrophysiological changes induced by chronic high-altitude hypoxia in adult rat heart. AB - Recent studies indicate that regression of left ventricular hypertrophy normalizes membrane ionic current abnormalities. This work was designed to determine whether regression of right ventricular hypertrophy induced by permanent high-altitude exposure (4,500 m, 20 days) in adult rats also normalizes changes of ventricular myocyte electrophysiology. According to the current data, prolonged action potential, decreased transient outward current density, and increased inward sodium/calcium exchange current density normalized 20 days after the end of altitude exposure, whereas right ventricular hypertrophy evidenced by both the right ventricular weight-to-heart weight ratio and the right ventricular free wall thickness measurement normalized 40 days after the end of altitude exposure. This morphological normalization occurred at both the level of muscular tissue, as shown by the decrease toward control values of some myocyte parameters (perimeter, capacitance, and width), and the level of the interstitial collagenous connective tissue. In the chronic high-altitude hypoxia model, the regression of right ventricular hypertrophy would not be a prerequisite for normalization of ventricular electrophysiological abnormalities. PMID- 11893583 TI - Preconditioning prevents alterations in cardiac SR gene expression due to ischemia-reperfusion. AB - We have previously shown that ischemic preconditioning (IP) improves cardiac performance and sarcoplasmic reticulum (SR) function in hearts subjected to ischemia-reperfusion (I/R). In this study, we examined the effect of IP on I/R induced changes in gene expression for SR proteins such as the Ca(2+) release channel, Ca(2+) pump ATPase, phospholamban, and calsequestrin in the isolated rat heart. Normal isolated rat hearts exposed to three brief cycles of IP (5-min ischemia and 5-min reperfusion) exhibited a significant decrease in the transcript levels of SR genes. Nonpreconditioned I/R hearts when subjected to 30 min ischemia and 30-min reperfusion showed a marked decrease in mRNA levels for the SR proteins compared with normal hearts; this decrease was attenuated by preconditioning. Although hearts subjected to Ca(2+) paradox (CP) have been shown to exhibit intracellular Ca(2+) overload and SR dysfunction like those in I/R hearts, virtually nothing is known regarding the effect of CP on cardiac SR gene expression. Accordingly, CP (5-min Ca(2+)-free perfusion and 30-min reperfusion with normal medium) was observed to produce dramatic changes in SR gene expression, and the heart failed to contract; these alterations were attenuated by IP. Our results show that 1) both I/R and CP depress SR gene expression in the normal heart, 2) IP attenuates I/R- and CP-induced depression in cardiac function and SR gene expression, and 3) intracellular Ca(2+) overload may play a role in depressing SR gene expression in both I/R and CP hearts. PMID- 11893584 TI - Hypoxia in the thymus: role of oxygen tension in thymocyte survival. AB - Our previous studies using oxygen microelectrodes showed that the thymus is grossly hypoxic under normal physiological conditions. We now have investigated how oxygen tension affects the thymus at the cellular and molecular level. Adducts of the hypoxia marker drug pimonidazole accumulated in foci within the cortex and medulla and at the corticomedullary junction, consistent with the presence of widespread cellular hypoxia in the normal thymus. Hypoxia-associated pimonidazole accumulation was decreased but not abrogated by oxygen administration. Genes previously reported to be induced by hypoxia were expressed at baseline levels in the normal thymus, indicating that physiological adaptation to hypoxia occurred. Despite changes in thymus size and cellularity, thymic PO(2) did not change with age. Combined assays for hypoxia and cell death showed that hypoxia achieved using either hypoxic gas mixtures or high-density culture in normoxia decreased spontaneous thymocyte apoptosis in vitro. Taken together, these data suggest that regulatory mechanisms exist to maintain thymic cellular hypoxia in vivo and that oxygen tension may regulate thymocyte survival both in vitro and in vivo. PMID- 11893585 TI - Long-term inhibition of nitric oxide synthesis increases arterial thrombogenecity in rat carotid artery. AB - Reduced activity of endothelial nitric oxide (NO) may be involved in thrombus formation on atherosclerotic plaques, a major cause of acute coronary syndrome. However, mechanisms of such increase in arterial thrombogenecity have not been fully understood. We previously reported that long-term inhibition of NO synthesis by administration of N(G)-nitro-L-arginine methyl ester (L-NAME) causes hypertension and activates vascular tissue angiotensin-converting enzyme (ACE) activity. We used this model to investigate the mechanism by which long-term impairment of NO activity increases arterial thrombogenecity. We observed cyclic flow variations (CFVs), a reliable marker of platelet thrombi, after the production of stenosis of the carotid artery in rats treated with L-NAME for 4 wk. The thrombin antagonist argatroban suppressed the CFVs. The CFVs were detected in rats receiving L-NAME plus hydralazine but not in rats receiving L NAME plus an ACE inhibitor (imidapril). Treatment with the ACE inhibitor imidapril, but not with hydralazine, prevented L-NAME-induced increases in carotid arterial ACE activity and attenuated tissue factor expression. These results suggest that long-term inhibition of endothelial NO synthesis may increase arterial thrombogenecity at least in part through angiotensin II-induced induction of tissue factor and the resultant thrombin generation. These data provide a new insight as to how endothelial NO exhibits antithrombogenic properties of the endothelium. PMID- 11893587 TI - Long-chain fatty acids increase basal metabolism and depolarize mitochondria in cardiac muscle cells. AB - The effects of long-chain (LC) fatty acids on rate of heat production (heat rate) and mitochondrial membrane potential (DeltaPsi) of intact guinea pig cardiac muscle were investigated at 37 degrees C. Heat rate of ventricular trabeculae was measured with microcalorimetry, and DeltaPsi was monitored in isolated ventricular myocytes with either JC-1 or tetramethylrhodamine ethyl ester (TMRE). Methyl-beta-cyclodextrin was used as fatty acid carrier. Application of 400 microM oleate or linoleate increased resting heat rate by approximately 30% and approximately 25%, respectively. When LC fatty acid was supplied as sole metabolic substrate, resting heat rate was decreased by 3-mercaptopropionic acid. In TMRE-loaded myocytes, neither 40-80 microM oleate nor 40 microM linoleate affected DeltaPsi. At a higher concentration (400 microM) both oleate and linoleate increased TMRE fluorescence by approximately 20% of maximum, obtained using 2,4-dinitrophenol (100 microM), indicating a depolarization of the inner mitochondrial membrane. We conclude that LC fatty acids, at sufficiently high concentration, increase heat rate and decrease DeltaPsi in intact cardiac muscle, consistent with a protonophoric uncoupling action. These effects may contribute to the high metabolic rate after reperfusion of postischemic myocardium. PMID- 11893588 TI - Validation of OPS imaging for microvascular measurements during isovolumic hemodilution and low hematocrits. AB - Orthogonal polarization spectral (OPS) imaging is a new technique that can be used to visualize the microcirculation with reflected light. It uses hemoglobin absorption to visualize the red blood cells (RBCs). Thus the method could fail at low hematocrit (Hct). The aim of this study was to validate OPS imaging for quantitative measurements of diameter and functional capillary density (FCD) under conditions of hemodilution of varying degrees to achieve a wide range of Hcts. The validation was performed in the dorsal skinfold chamber of nine awake Syrian golden hamsters. Measurements of vessel diameter and FCD were performed off-line using Cap-Image on video sequences captured using OPS imaging and standard intravital fluorescence microscopy at baseline, 85, 70, 55, and 40% of the initial Hct. For hemodilution, isovolumic exchange of blood for 6% Dextran 60 was performed. Bland-Altman plots for the vessel diameter and FCD show good agreement between the two methods for both parameters at all studied Hcts. As expected, there was a systematic bias of approximately 4 microm in the diameter measurements since the RBC column was measured and not the intravascular diameter. In conclusion, OPS imaging can be used to measure diameter and FCD at a wide range of Hcts. PMID- 11893586 TI - Cerebral microvascular changes in permeability and tight junctions induced by hypoxia-reoxygenation. AB - Cerebral microvessel endothelial cells that form the blood-brain barrier (BBB) have tight junctions (TJ) that are critical for maintaining brain homeostasis and low permeability. Both integral (claudin-1 and occludin) and membrane-associated zonula occluden-1 and -2 (ZO-1 and ZO-2) proteins combine to form these TJ complexes that are anchored to the cytoskeletal architecture (actin). Disruptions of the BBB have been attributed to hypoxic conditions that occur with ischemic stroke, pathologies of decreased perfusion, and high-altitude exposure. The effects of hypoxia and posthypoxic reoxygenation in cerebral microvasculature and corresponding cellular mechanisms involved in disrupting the BBB remain unclear. This study examined hypoxia and posthypoxic reoxygenation effects on paracellular permeability and changes in actin and TJ proteins using primary bovine brain microvessel endothelial cells (BBMEC). Hypoxia induced a 2.6-fold increase in [(14)C]sucrose, a marker of paracellular permeability. This effect was significantly reduced (~58%) with posthypoxic reoxygenation. After hypoxia and posthypoxic reoxygenation, actin expression was increased (1.4- and 2.3-fold, respectively). Whereas little change was observed in TJ protein expression immediately after hypoxia, a twofold increase in expression was seen with posthypoxic reoxygenation. Furthermore, immunofluorescence studies showed alterations in occludin, ZO-1, and ZO-2 protein localization during hypoxia and posthypoxic reoxygenation that correlate with the observed changes in BBMEC permeability. The results of this study show hypoxia-induced changes in paracellular permeability may be due to perturbation of TJ complexes and that posthypoxic reoxygenation reverses these effects. PMID- 11893589 TI - Contribution of laminar myofiber architecture to load-dependent changes in mechanics of LV myocardium. AB - The ventricular myocardium consists of a syncytium of myocytes organized into branching, transmurally oriented laminar sheets approximately four cells thick. When systolic deformation is expressed in an axis system determined by the anatomy of the laminar architecture, laminar sheets of myocytes shear and laterally extend in an approximately radial direction. These deformations account for ~90% of normal systolic wall thickening in the left ventricular free wall. In the present study, we investigated whether the changes in systolic and diastolic function of the sheets were sensitive to alterations in systolic and diastolic load. Our results indicate that there is substantial reorientation of the laminar architecture during systole and diastole. Moreover, this reorientation is both site and load dependent. Thus as end-diastolic pressure is increased and the left ventricular wall thins, sheets shorten and rotate away from the radial direction due to transverse shearing, opposite of what occurs in systole. Both mechanisms of thickening contribute substantially to normal left ventricular wall function. Whereas the relative contributions of shear and extension are comparable at the base, sheet shear is the predominant factor at the apex. The magnitude of shortening/extension and shear increases with preload and decreases with afterload. These findings underscore the essential contribution of the laminar myocardial architecture for normal ventricular function throughout the cardiac cycle. PMID- 11893590 TI - Increased expression of alternatively spliced dominant-negative isoform of SRF in human failing hearts. AB - Serum response factor (SRF) has been shown to play a key role in cardiac cell growth and muscle gene regulation. To understand the role of SRF in heart failure, we compared its expression pattern between control and failing human heart samples. Western blot analysis of control samples showed expression of four different isoforms of SRF, with ~67-kDa full-length SRF being the predominant isoform. Interestingly, in failing hearts we found robust expression of a low molecular-mass (~52 kDa) SRF isoform, accompanied by decreased expression of full length SRF. By RT-PCR and Southern blot analyses, we characterized this ~52-kDa SRF isoform as being encoded by an alternatively spliced form of SRF lacking exons 4 and 5 of the SRF primary RNA transcript (SRF-Delta4,5 isoform). We cloned SRF-Delta4,5 cDNA and showed that overexpression of this isoform into cells inhibits SRF-dependent activation of cardiac muscle genes. These results suggest that expression of SRF-Delta4,5 in failing hearts may in part contribute to impaired cardiac gene expression and consequently to the pathogenesis of heart failure. PMID- 11893591 TI - Mathematical analysis of dynamics of cardiac memory and accommodation: theory and experiment. AB - Decreasing the slope of the dynamic, but not conventional, restitution curves is antifibrillatory. Cardiac memory/accommodation underlies the difference. We measured diastolic interval (DI) and action potential duration (APD) in epicardial, endocardial, and Purkinje tissue from eight dogs. Consecutive 100 stimulus trains were given to study transitions between basic cycle lengths (BCL) ranging from 400 to 1,300 ms. (DI,APD) pairs aligned immediately on the line DI + APD = BCL (64/67) or oscillated (3/67). The shifting effect of up to 10 extrastimuli on restitution curves was also measured. These curves were fit with the equation APD = alpha + beta exp(-DI/tau), where alpha is asymptote, beta is drop, and tau is time constant. Linear regression of the parameters against the number of extrastimuli showed that premature and postmature stimuli decreased and increased alpha and beta and increased and decreased tau, respectively. Analysis of a mathematical model treating memory as an exponentially decreasing shift of restitution curves shows that oscillatory DI,APD is expected with large DeltaBCL, steep restitution slope, or increased cardiac accommodation. The model explains phase shifts and suggests a common mechanism for Purkinje and myocardial electrical alternans. PMID- 11893592 TI - Gap junction-dependent and -independent EDHF-type relaxations may involve smooth muscle cAMP accumulation. AB - We have compared the mechanisms that contribute to endothelium-derived hyperpolarizing factor (EDHF)-type responses induced by ACh and the Ca(2+) ionophore A-23187 in the rabbit iliac artery. Relaxations to both agents were associated with ~1.5-fold elevations in smooth muscle cAMP levels and were attenuated by the adenylyl cyclase inhibitor 2',5'-dideoxyadenosine (DDA) and potentiated by the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). Mechanical responses were inhibited by coadministration of the Ca(2+) activated K(+) channel blockers apamin and charybdotoxin, both in the absence and presence of IBMX, but were unaffected by blockade of ATP-sensitive K(+) channels with the sulphonylurea glibenclamide. Relaxations and elevations in cAMP evoked by ACh were abolished by 18alpha-glycyrrhetinic acid, which disrupts gap junction plaques, whereas the corresponding responses to A-23187 were unaffected by this agent. Consistently, in "sandwich" bioassay experiments, A-23187, but not ACh, elicited extracellular release of a factor that evoked relaxations that were inhibited by DDA and potentiated by IBMX. These findings provide evidence that EDHF-type relaxations of rabbit iliac arteries evoked by ACh and A-23187 depend on cAMP accumulation in smooth muscle, but involve signaling via myoendothelial gap junctions and the extracellular space, respectively. PMID- 11893593 TI - 20-HETE contributes to the acute fall in cerebral blood flow after subarachnoid hemorrhage in the rat. AB - This study examined the effects of blocking the formation of 20 hydroxyeicosatetraenoic acid (20-HETE) on the acute fall in cerebral blood flow after subarachnoid hemorrhage (SAH) in the rat. In vehicle-treated rats, regional cerebral blood flow (rCBF) measured with laser-Doppler flowmetry fell by 30% 10 min after the injection of 0.3 ml of arterial blood into the cisterna magna, and it remained at this level for 2 h. Pretreatment with inhibitors of the formation of 20-HETE, 17-octadecynoic acid (17-ODYA; 1.5 nmol intrathecally) and N-hydroxy N'-(4-butyl-2-methylphenyl)formamidine (HET0016; 10 mg/kg iv), reduced the initial fall in rCBF by 40%, and rCBF fully recovered 1 h after induction of SAH. The concentration of 20-HETE in the cerebrospinal fluid rose from 12 +/- 2 to 199 +/- 17 ng/ml after SAH in vehicle-treated rats. 20-HETE levels averaged only 15 +/- 11 and 39 +/- 13 ng/ml in rats pretreated with 17-ODYA or HET0016, respectively. HET0016 selectively inhibited the formation of 20-HETE in rat renal microsomes with an IC(50) of <15 nM and human recombinant CYP4A11, CYP4F2, and CYP4F3 enzymes with an IC(50) of 42, 125, and 100 nM, respectively. These results indicate that 20-HETE contributes to the acute fall in rCBF after SAH in rats. PMID- 11893595 TI - Renin. PMID- 11893594 TI - Acute exercise reduces the response to colon distension in T(5) spinal rats. AB - Individuals with spinal cord injuries above thoracic level 6 (T(6)) experience life-threatening bouts of hypertension, termed autonomic dysreflexia (AD). AD is mediated by peripheral alpha-adrenergic receptor supersensitivity as well as a reorganization of spinal pathways controlling sympathetic preganglionic neurons. A single bout of dynamic exercise may be a safe therapeutic approach to reduce the severity of AD because mild-to-moderate dynamic exercise reduces postexercise alpha-adrenergic receptor responsiveness, lowers postexercise sympathetic nerve activity, and reduces the postexercise response to stress. Therefore, this study was designed to test the hypothesis that mild-to-moderate dynamic exercise attenuates the postexercise response to colon distension (mechanism to elicit AD). To test this hypothesis, six male Wistar rats (406 +/- 23 g), 5 wk post-T(5) spinal cord transection, were instrumented with an arterial catheter. After recovery, the response to graded colon distension (10, 30, 50, and 80 mmHg, in random order) was determined before and after a single bout of mild-to-moderate dynamic exercise (9-12 m/min, 0% grade for 40 min). After exercise, the pressor response to graded colon distension was significantly attenuated (preexercise change: 2 +/- 1, 9 +/- 1, 14 +/- 1, and 24 +/- 2 vs. postexercise change: 2 +/- 1, 2 +/- 1, 9 +/- 1, and 12 +/- 3 mmHg). Thus acute exercise is a safe, therapeutic approach to reduce the severity of AD in paraplegic subjects. PMID- 11893596 TI - The natriuretic peptide system in eels: a key endocrine system for euryhalinity? AB - The natriuretic peptide system of a euryhaline teleost, the Japanese eel (Anguilla japonica), consists of three types of hormones [atrial natriuretic peptide (ANP), ventricular natriuretic peptide (VNP), and C-type natriuretic peptide (CNP)] and four types of receptors [natriuretic peptide receptors (NPR) A, -B, -C, and -D]. Although ANP is recognized as a volume-regulating hormone that extrudes both Na(+) and water in mammals, ANP more specifically extrudes Na(+) in eels. Accumulating evidence shows that ANP is secreted in response to hypernatremia and acts to inhibit the uptake and to stimulate the excretion of Na(+) but not water, thereby promoting seawater (SW) adaptation. In fact, ANP is secreted immediately after transfer of eels to SW and ameliorates sudden increases in plasma Na(+) concentration through inhibition of drinking and intestinal absorption of NaCl. ANP also stimulates the secretion of cortisol, a long-acting hormone for SW adaptation, whereas ANP itself disappears quickly from the circulation. Thus ANP is a primary hormone responsible for the initial phase of SW adaptation. By contrast, CNP appears to be a hormone involved in freshwater (FW) adaptation. Recent data show that the gene expression of CNP and its specific receptor, NPR-B, is much enhanced in FW eels. In fact, CNP infusion increases (22)Na uptake from the environment in FW eels. These results show that ANP and CNP, despite high sequence identity, have opposite effects on salinity adaptation in eels. This difference apparently originates from the difference in their specific receptors, ANP for NPR-A and CNP for NPR-B. VNP may compensate the effects of ANP and CNP for adaptation to respective media, because it has high affinity to both receptors. On the basis of these data, the authors suggest that the natriuretic peptide system is a key endocrine system that allows this euryhaline fish to adapt to diverse osmotic environments, particularly in the initial phase of adaptation. PMID- 11893597 TI - Enhanced vascular effects of the Ca(2+) channel agonist Bay K 8644 in pregnant rabbits. AB - Hemodynamic studies were performed to determine if blunting of vascular pressor responsiveness to vasoconstrictors during pregnancy may be due to impaired L-type voltage-dependent calcium channels (L-VDCC). Bay K 8644 (BAY), an L-VDCC agonist, was infused in pregnant and nonpregnant anesthetized rabbits (10, 20, 40, and 60 microg/kg) and pregnant and nonpregnant conscious, chronically instrumented (conscious) rabbits (10, 25, and 50 microg/kg). BAY infusions resulted in greater elevation of mean arterial pressure in both anesthetized pregnant (n = 6) vs. nonpregnant (n = 6) (P < 0.05) and conscious pregnant (n = 10) vs. nonpregnant (n = 10) rabbits (P < 0.05). Fractional increase over baseline of total peripheral resistance index was greater in pregnant (36 +/- 5 to 78 +/- 14%) vs. nonpregnant rabbits (14 +/- 4 to 52 +/- 6%) (P < 0.02). Cardiac output index did not differ. There was a single high-affinity L-VDCC antagonist aortic binding site with similar number and affinity in pregnant (n = 7) and nonpregnant (n = 7) rabbits. In conclusion, stimulation of L-VDCC induces greater pressor responses in pregnant rabbits with heightened peripheral vasoconstriction. This does not appear to be due to a change in L-VDCC receptor parameters. PMID- 11893598 TI - Postexercise alpha-adrenergic receptor hyporesponsiveness in hypertensive rats is due to nitric oxide. AB - We tested the hypothesis that a single bout of dynamic exercise produces a postexercise hypotension (PEH) and alpha(1)-adrenergic receptor hyporesponsiveness in spontaneously hypertensive rats (SHR). The postexercise alpha(1)-adrenergic receptor hyporesponsiveness is due to an enhanced buffering of vasoconstriction by nitric oxide. Male (n = 8) and female (n = 5) SHR were instrumented with a Doppler ultrasonic flow probe around the femoral artery. Distal to the flow probe, a microrenathane catheter was inserted into a branch of the femoral artery for the infusion of the alpha(1)-adrenergic receptor agonist phenylephrine (PE). A microrenathane catheter was inserted into the descending aorta via the left common carotid artery for measurements of arterial pressure (AP) and heart rate. Dose-response curves to PE (3.8 x 10(-3) - 1.98 x 10( 2)microg/kHz) were generated before and after a single bout of dynamic exercise. Postexercise AP was reduced in male (13 +/- 3 mmHg) and female SHR (18 +/- 7 mmHg). Postexercise vasoconstrictor responses to PE were reduced in males due to an enhanced influence of nitric oxide. However, in females, postexercise vasoconstrictor responses to PE were not altered. Results suggest that nitric oxide- mediated alpha(1)-adrenergic receptor hyporesponsiveness contributes to PEH in male but not female SHR. PMID- 11893599 TI - Multiple dilator pathways in skeletal muscle contraction-induced arteriolar dilations. AB - To determine whether nitric oxide (NO), adenosine (Ado) receptors, or ATP sensitive potassium (K(ATP)) channels play a role in arteriolar dilations induced by muscle contraction, we used a cremaster preparation in anesthetized hamsters in which we stimulated four to five muscle fibers lying perpendicular to a transverse arteriole (maximal diameter approximately 35-65 microm). The diameter of the arteriole at the site of overlap of the stimulated muscle fibers (the local site) and at a remote site approximately 1,000 microm upstream (the upstream site) was measured before, during, and after muscle contraction. Two minutes of 4-Hz muscle stimulation (5-15 V, 0.4 ms) produced local and upstream dilations of 19 +/- 1 and 10 +/- 1 microm, respectively. N(omega)-nitro-L arginine (10(-4) M; NO synthase inhibitor), xanthine amine congener (XAC; 10(-6) M; Ado A(1), A(2A), and A(2B) receptor antagonist), or glibenclamide (Glib; 10( 5) M; K(ATP) channel inhibitor) superfused over the preparation attenuated the local dilation (by 29.7 +/- 12.7, 61.8 +/- 9.0, and 51.9 +/- 14.9%, respectively), but only XAC and Glib attenuated the upstream dilation (by 68.9 +/ 6.8 and 89.1 +/- 6.4%, respectively). Furthermore, only Glib, when applied to the upstream site directly, attenuated the upstream dilation (48.1 +/- 9.1%). Neither XAC nor Glib applied directly to the arteriole between the local and the upstream sites had an effect on the magnitude of the upstream dilation. We conclude that NO, Ado receptors, and K(ATP) channels are involved in the local dilation initiated by contracting muscle and that both K(ATP) channels and Ado receptor stimulation, but not NO, play a role in the manifestation of the dilation at the upstream site. PMID- 11893600 TI - Cytokine-mediated downregulation of vasopressin V(1A) receptors during acute endotoxemia in rats. AB - The reduced pressure response to vasopressin during acute sepsis has directed our interest to the regulation of vasopressin V(1A) receptors. Rats were injected with lipopolysaccharide for induction of experimental gram-negative sepsis. V(1A) receptor gene expression was downregulated in the liver, lung, kidney, and heart during endotoxemia. Inasmuch as the concentrations of proinflammatory cytokines such as interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma were highly increased during sepsis, the influence of these cytokines on V(1A) receptor expression was investigated in primary cultures of hepatocytes and in the aortic vascular smooth muscle cell line A7r5. V(1A) receptor expression was downregulated by the cytokines in a nitric oxide-independent manner. Blood pressure dose-response studies after injection of endotoxin showed a diminished responsiveness to the selective V(1) receptor agonist Phe(2),Ile(3),Orn(8) vasopressin. Our data show that sepsis causes a downregulation of V(1A) receptors and suggest that this effect is likely mediated by proinflammatory cytokines. We propose that this downregulation of V(1A) receptors contributes to the attenuated responsiveness of blood pressure in response to vasopressin and, therefore, contributes to the circulatory failure in septic shock. PMID- 11893601 TI - Behavioral dysfunction, brain oxidative stress, and impaired mitochondrial electron transfer in aging mice. AB - Behavioral tests, tightrope success, and exploratory activity in a T maze were conducted with male and female mice for 65 wk. Four groups were defined: the lower performance slow males and slow females and the higher performance fast males and fast females. Fast females showed the longest life span and the highest performance, and slow males showed the lowest performance and the shortest life span. Oxidative stress and mitochondrial electron transfer activities were determined in brain of young (28 wk), adult (52 wk), and old (72 wk) mice in a cross-sectional study. Brain thiobarbituric acid reactive substances (TBARS) were increased by 50% in old mice and were approximately 15% higher in males than in females and in slow than in fast mice. Brain Cu,Zn-superoxide dismutase (SOD) activity was increased by 52% and Mn-SOD by 108% in old mice. The activities of mitochondrial enzymes NADH-cytochrome c reductase, cytochrome oxidase, and citrate synthase were decreased by 14-58% in old animals. The cumulative toxic effects of oxyradicals are considered the molecular mechanism of the behavioral deficits observed on aging. PMID- 11893602 TI - Actions of a novel synthetic natriuretic peptide on hemodynamics and ventricular function in the dog. AB - Dendroaspis natriuretic peptide (DNP) is a recently discovered peptide with structural similarity to known natriuretic peptides. DNP has been shown to possess potent renal actions. Our objectives were to define the acute hemodynamic actions of DNP in normal anesthetized dogs and the acute effects of DNP on left ventricular (LV) function in conscious chronically instrumented dogs. In anesthetized dogs, DNP, but not placebo, decreased mean arterial pressure (141 +/ 6 to 109 +/- 7 mmHg, P < 0.05) and pulmonary capillary wedge pressure (5.8 +/- 0.3 to 3.4 +/- 0.2 mmHg, P < 0.05). Cardiac output decreased and systemic vascular resistance increased with DNP and placebo. DNP-like immunoreactivity and guanosine 3',5'-cyclic monophosphate concentration increased without changes in other natriuretic peptides. In conscious dogs, DNP decreased LV end-systolic pressure (120 +/- 7 to 102 +/- 6 mmHg, P < 0.05) and volume (32 +/- 6 to 28 +/- 6 ml, P < 0.05) and LV end-diastolic volume (38 +/- 5 to 31 +/- 4 ml, P < 0.05) but not arterial elastance. LV end-systolic elastance increased (6.1 +/- 0.7 to 7.4 +/- 0.6 mmHg/ml, P < 0.05), and Tau decreased (31 +/- 2 to 27 +/- 1 ms, P < 0.05). The effects on hemodynamics, LV function, and second messenger generation suggest synthetic DNP may have a role as a cardiac unloading and lusitropic peptide. PMID- 11893603 TI - Trigeminal reflex regulation of the glottis depends on central glycinergic inhibition in the rat. AB - In an unanesthetized decerebrate in situ arterially perfused brain stem preparation of mature rat, strychnine (0.05-0.2 microM) blockade of glycine receptors caused postinspiratory glottal constriction to occur earlier, shifting from early expiration to inspiration. This resulted in a paradoxical inspiratory related narrowing of the upper airway. Stimulation of the trigeminal ethmoidal nerve (EN5; 20 Hz, 100 micros, 0.5-2 V) evoked a diving response, which included a reflex apnea, glottal constriction, and bradycardia. After strychnine administration, this pattern was converted to a maintained phrenic nerve discharge and a reduced glottal constriction that was interrupted intermittently by transient abductions. The onset of firing of postinspiratory neurons shifted from early expiration into neural inspiration in the presence of strychnine, but neurons maintained their tonic activation during EN5 stimulation, as observed during control. Inspiratory neurons that were hyperpolarized by EN5 stimulation in control conditions were powerfully excited after loss of glycinergic inhibition. Thus the integrity of glycinergic inhibition within the pontomedullary respiratory network is critical for the coordination of cranial and spinal motor outflows during eupnea but also for protective reflex regulation of the upper airway. PMID- 11893604 TI - Reduced endogenous GABA-mediated inhibition in the PVN on renal nerve discharge in rats with heart failure. AB - One characteristic of heart failure (HF) is increased sympathetic activation. The paraventricular nucleus (PVN) of the hypothalamus (involved in control of sympathetic outflow) has been shown to have increased neuronal activation during HF. This study examined the influence of endogenous GABA input (inhibitory in nature) into the PVN on renal sympathetic nerve discharge (RSND), arterial blood pressure (BP), and heart rate (HR) in rats with HF induced by coronary artery ligation. In alpha-chloralose- and urethane-anesthetized rats, microinjection of bicuculline (a GABA antagonist) into the PVN produced a dose-dependent increase in RSND, BP, and HR in both sham-operated control and HF rats. Bicuculline attenuated the increase in RSND and BP in HF rats compared with control rats. Alternatively, microinjection of the GABA agonist muscimol produced a dose dependent decrease in RSND, BP, and HR in both control and HF rats. Muscimol was also less effective in decreasing RSND, BP, and HR in HF rats than in control rats. These results suggest that endogenous GABA-mediated input into the PVN of rats with HF is less effective in suppressing RSND and BP compared with control rats. This is partly due to the post-release actions of GABA, possibly caused by altered function of post-synaptic GABA receptors in the PVN of rats with HF. Reduced GABA-mediated inhibition in the PVN may contribute to increased sympathetic outflow, which is commonly observed during HF. PMID- 11893605 TI - Proteasome inhibitors induce heat shock response and increase IL-6 expression in human intestinal epithelial cells. AB - In previous studies, the heat shock response, induced by hyperthermia or sodium arsenite, increased interleukin (IL)-6 production in intestinal mucosa and cultured human enterocytes. A novel way to induce the heat shock response, documented in other cell types, is treatment with proteasome inhibitors. It is not known if proteasome inhibition induces heat shock in enterocytes or influences IL-6 production. Here we tested the hypothesis that treatment of cultured Caco-2 cells, a human intestinal epithelial cell line, with proteasome inhibitors induces the heat shock response and stimulates IL-6 production. Treatment of Caco-2 cells with one of the proteasome inhibitors MG-132 or lactacystin activated the transcription factor heat shock factors (HSF)-1 and -2 and upregulated cellular levels of the 72-kDa heat shock protein HSP-72. The same treatment resulted in increased gene and protein expression of IL-6, a response that was blocked by quercetin. Additional experiments revealed that the IL-6 gene promoter contains a HSF-responsive element and that the IL-6 gene may be regulated by the heat shock response. The present results suggest that proteasome inhibition induces heat shock response and IL-6 production in enterocytes and that IL-6 may be a heat shock-responsive gene, at least under certain circumstances. The observations are important considering the multiple biological roles of IL-6, both locally in the gut mucosa and systemically, and considering recent proposals in the literature to use proteasome inhibitors in the clinical setting to induce the heat shock response. PMID- 11893606 TI - Blood flows and nutrient uptakes in growth-restricted pregnancies induced by overnourishing adolescent sheep. AB - To establish physiological mechanisms for fetal growth restriction in pregnant adolescent ewes we studied uterine, fetal, and uteroplacental metabolism in ewes offered a high (n = 12) or moderate (n = 10) dietary intake. High intakes decreased placental (226 vs. 414 g, P < 0.001) and fetal weight (3,323 vs. 4,626 g, P < 0.01). Uterine blood flow was reduced absolutely (-36%) but proportional to conceptus weight; umbilical blood flow was reduced absolutely (-37%) and per fetal weight (-15%). Uterine oxygen uptake was decreased per conceptus weight ( 14%); there was no change in fetal weight oxygen consumption. Uteroplacental oxygen consumption and clearance were reduced proportional to weight. Similar changes were measured for glucose fluxes and fetal glucose concentration; fetal insulin concentration was reduced. In this model of fetal growth restriction, therefore, maintenance of fetal weight-specific glucose and oxygen consumption rates are producing relative hypoglycemia and hypoxemia. This indicates that increased fetal glucose clearance and/or insulin sensitivity may be operating as compensatory mechanisms to preserve normal fetal metabolism while fetal growth is sacrificed. PMID- 11893607 TI - Hemodynamic and autonomic correlates of postexercise hypotension in patients with mild hypertension. AB - We investigated the interplay of neural and hemodynamic mechanisms in postexercise hypotension (PEH) in hypertension. In 15 middle-aged patients with mild essential hypertension, we evaluated blood pressure (BP), cardiac output (CO), total peripheral resistance (TPR), forearm (FVR) and calf vascular resistance (CVR), and autonomic function [by spectral analysis of R-R interval and BP variabilities and spontaneous baroreflex sensitivity (BRS)] before and after maximal exercise. Systolic and diastolic BP, TPR, and CVR were significantly reduced from baseline 60-90 min after exercise. CO, FVR, and HR were unchanged. The low-frequency (LF) component of BP variability increased significantly after exercise, whereas the LF component of R-R interval variability was unchanged. The overall change in BRS was not significant after exercise vs. baseline, although a significant, albeit small, BRS increase occurred in response to hypotensive stimuli. These findings indicate that in hypertensive patients, PEH is mediated mainly by a peripheral vasodilation, which may involve metabolic factors linked to postexercise hyperemia in the active limbs. The vasodilator effect appears to override a concomitant, reflex sympathetic activation selectively directed to the vasculature, possibly aimed to counter excessive BP decreases. The cardiac component of arterial baroreflex is reset during PEH, although the baroreflex mechanisms controlling heart period appear to retain the potential for greater opposition to hypotensive stimuli. PMID- 11893608 TI - Unloading arterial baroreceptors causes neurogenic hypertension. AB - We developed a new model to examine the role of arterial baroreceptors in the long-term control of mean arterial pressure (MAP) in dogs. Baroreceptors in the aortic arch and one carotid sinus were denervated, and catheters were implanted in the descending aorta and common carotid arteries. MAP and carotid sinus pressure (CSP) averaged 104 +/- 2 and 102 +/- 2 mmHg (means +/- 1 SE), respectively, during a 5-day control period. Baroreceptor unloading was induced by ligation of the common carotid artery proximal to the innervated sinus (n = 6 dogs). MAP and CSP averaged 127 +/- 7 and 100 +/- 3 mmHg, respectively, during the 7-day period of baroreceptor unloading. MAP was significantly elevated (P < 0.01) compared to control, but CSP was unchanged. Heart rate and plasma renin activity increased significantly in response to baroreceptor unloading. Removal of the ligature to restore normal flow through the carotid resulted in normalization of all variables. Ligation of the carotid below a denervated sinus (n = 4) caused a significant decrease in CSP but no systemic hypertension. These results indicate that chronic unloading of carotid baroreceptors can produce neurogenic hypertension and provide strong evidence that arterial baroreceptors are involved in the long-term control of blood pressure. PMID- 11893609 TI - Periodic arousal from hibernation is necessary for initiation of immune responses in ground squirrels. AB - Golden-mantled ground squirrels (Spermophilus lateralis) undergo seasonal hibernation during which core body temperature (T(b)) values are maintained 1-2 degrees C above ambient temperature. Hibernation is not continuous. Squirrels arouse at approximately 7-day intervals, during which T(b) increases to 37 degrees C for approximately 16 h; thereafter, they return to hibernation and sustain low T(b)s until the next arousal. Over the course of the hibernation season, arousals consume 60-80% of a squirrel's winter energy budget, but their functional significance is unknown and disputed. Host-defense mechanisms appear to be downregulated during the hibernation season and preclude normal immune responses. These experiments assessed immune function during hibernation and subsequent periodic arousals. The acute-phase response to bacterial lipopolysaccharide (LPS) was arrested during hibernation and fully restored on arousal to normothermia. LPS injection (ip) resulted in a 1-1.5 degrees C fever in normothermic animals that was sustained for > 8 h. LPS was without effect in hibernating squirrels, neither inducing fever nor provoking arousal, but a fever did develop several days later, when squirrels next aroused from hibernation; the duration of this arousal was increased sixfold above baseline values. Intracerebroventricular infusions of prostaglandin E(2) provoked arousal from hibernation and induced fever, suggesting that neural signaling pathways that mediate febrile responses are functional during hibernation. Periodic arousals may activate a dormant immune system, which can then combat pathogens that may have been introduced immediately before or during hibernation. PMID- 11893610 TI - Aging and assessment of physiological strain during exercise-heat stress. AB - The purpose of this study was to evaluate the physiological strain index (PSI) for different age groups during exercise-heat stress (EHS). PSI was applied to three different databases. First, from young and middle-age men (21 +/- 2 and 46 +/- 5 yr, respectively) matched (n = 9 each, P > 0.05) for maximal aerobic power. Subjects were heat acclimated by daily treadmill walking for two 50-min bouts separated by 10-min rest for 10 days in a hot-dry environment [49 degrees C, 20% relative humidity (RH)]. The second database involved a group (n = 8) of young (YA) and a group (n = 7) of older (OA) men (26 +/- 1 and 69 +/- 1 yr, respectively) who underwent 16 wk of aerobic training and two control groups (n = 7 each) who were matched for age to YA and OA. These four groups performed EHS at 36 degrees C, 40% RH on a cycle ergometer for 60 min at 60% maximal aerobic power before and after training. The third database was obtained from three groups of postmenopausal women and a group of 10 men. Two groups of women (n = 8 each) were undergoing hormone replacement therapy, estrogen or estrogen plus progesterone, and the third group (n = 9) received no hormone replacement. Subjects were over 50 yr and performed the same EHS: exercising at 36 degrees C, 40% RH on a cycle ergometer for 60 min. PSI assessed the strain for all three databases and reported differences were significant at P < 0.05. This index rated the strain in rank order, whereas the postacclimation and posttraining groups were assessed as having less strain than the preacclimation and pretraining groups. Furthermore, middle-aged women on estrogen replacement therapy had less strain than estrogen + progesterone and no hormone therapy. PSI evaluation was extended for men and women of different ages (50-70 yr) during acute EHS, heat acclimation, after aerobic training, and inclusive of women undergoing hormone replacement therapy. PMID- 11893611 TI - Hypertension in L-NAME-treated diabetic rats depends on an intact sympathetic nervous system. AB - We demonstrated previously that induction of diabetes in rats that were treated chronically with the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) causes a severe, progressive increase in mean arterial pressure. This study tested the role of the sympathetic nervous system in that response. Rats were instrumented with chronic artery and vein catheters and assigned randomly to four diabetic groups pretreated with vehicle (D), L-NAME (D+L), the alpha(1)- and beta-adrenergic receptor antagonists terazosin and propranolol (D+B), or L-NAME, terazosin, and propranolol (D+LB). After baseline measurements were taken, rats were pretreated; 6 days later, streptozotocin was administered and 3 wk of diabetes ensued. D+L rats had a marked, progressive increase in arterial pressure that by day 20 was approximately 60 mmHg greater than in D rats. The pressor response to L-NAME was significantly attenuated in diabetic rats cotreated with adrenergic blockers. During week 1 of diabetes, plasma renin activity (PRA) increased and then returned to control levels in D rats. PRA increased progressively in D+L rats, and chronic adrenergic receptor blockade restored the biphasic renin response in D+LB rats. These results suggest that the sympathetic nervous system may be involved in the hypertensive response to onset of diabetes in L-NAME-treated rats, possibly through control of renin secretion. PMID- 11893612 TI - ATP-stimulated Ca(2+)-activated K(+) efflux pathway and differentiation of human placental cytotrophoblast cells. AB - The aim of this study was to determine whether extracellular ATP ([ATP](o)) stimulated a Ca(2+)-activated K(+) efflux in trophoblast cells that was dependent on extracellular Ca(2+) ([Ca(2+)](o)). Cytotrophoblast cells, isolated from human placenta, were examined following 18 h (relatively undifferentiated) and 66 h (multinucleate cells) of culture. Potassium efflux was measured using (86)Rb as a trace marker. Intracellular Ca(2+) ([Ca(2+)](i)) was examined by microfluorometry using fura 2. [ATP](o) significantly increased (86)Rb efflux to a peak that declined to control (18-h cells) or an elevated plateau (66-h cells) and was inhibited by 100 nM charybdotoxin. Removing [Ca(2+)](o) significantly reduced (86)Rb efflux in both groups as did application of 150 microM GdCl(3). [ATP](o) significantly increased [Ca(2+)](i) in both groups of cells. The response was reduced by removing [Ca(2+)](o) and applying 150 microM GdCl(3). For both (86)Rb efflux and microfluorometry experiments, the response to [ATP](o) was more dependent on [Ca(2+)](o) in 66-h cells compared with 18-h cells (approximately 70% greater). Cytotrophoblast cells exhibit an [ATP](o)-stimulated Ca(2+) activated K(+) efflux. The dependency of this pathway on [Ca(2+)](o) is greater in the 66-h multinucleate syncytiotrophoblast-like cells, suggesting that the mechanism for Ca(2+) entry may be altered during differentiation of trophoblast cells. PMID- 11893613 TI - Loss of circadian organization of sleep and wakefulness during hibernation. AB - We investigated circadian and homeostatic regulation of nonrapid eye movement (NREM) sleep in golden-mantled ground squirrels during euthermic intervals between torpor bouts. Slow-wave activity (SWA; 1-4 Hz) and sigma activity (10-15 Hz) represent the two dominant electroencephalographic (EEG) frequency components of NREM sleep. EEG sigma activity has a strong circadian component in addition to a sleep homeostatic component, whereas SWA mainly reflects sleep homeostasis [Dijk DJ and Czeisler CA. J Neurosci 15: 3526-3538, 1995; Dijk DJ, Shanahan TL, Duffy JF, Ronda JM, and Czeisler CA. J Physiol (Lond) 505: 851-858, 1997]. Animals maintained under constant conditions continued to display circadian rhythms in both sigma activity and brain temperature throughout euthermic intervals, whereas sleep and wakefulness showed no circadian organization. Instead, sleep and wakefulness were distributed according to a 6-h ultradian rhythm. SWA, NREM sleep bout length, and sigma activity responded homeostatically to the ultradian sleep-wake pattern. We suggest that the loss of sleep-wake consolidation in ground squirrels during the hibernation season may be related to the greatly decreased locomotor activity during the hibernation season and may be necessary for maintenance of multiday torpor bouts characteristic of hibernating species. PMID- 11893614 TI - Glomerulotubular balance, dietary protein, and the renal response to glycine in diabetic rats. AB - The glomerular filtration rate (GFR) normally increases during glycine infusion, which is a test of "renal reserve." Renal reserve is absent in diabetes mellitus. GFR increases after protein feeding because of increased tubular reabsorption, which reduces the signal for tubuloglomerular feedback (TGF). Dietary protein restriction normalizes some aspects of glomerular function in diabetes. Renal micropuncture was performed in rats 4-5 wk after diabetes was induced by streptozotocin to determine whether renal reserve is lost as a result of altered tubular function and activation of TGF, whether 10 days of dietary protein restriction could restore renal reserve, and whether this results from effects of glycine on the tubule. TGF activation was determined by locating single-nephron GFR (SNGFR) in the early distal tubule along the TGF curve. The TGF signal was determined from the ionic content of the early distal tubule. In nondiabetic rats, SNGFR in the early distal tubule increased during glycine infusion because of primary vasodilation augmented by increased tubular reabsorption, which stabilized the TGF signal. In diabetic rats, glycine reduced reabsorption, thereby activating TGF, which was largely responsible for the lack of renal reserve. In protein-restricted diabetic rats, the tubular response to glycine remained abnormal, but renal reserve was restored by a vascular mechanism. Glycine affects GFR directly and via the tubule. In diabetes, reduced tubular reabsorption dominates. In low-protein diabetes, the vascular effect is enhanced and overrides the effect of reduced tubular reabsorption. PMID- 11893615 TI - Decreased antigen-induced eicosanoid release in conjugated linoleic acid-fed guinea pigs. AB - This study investigated the capacity of conjugated linoleic acids (CLA) to reduce ex vivo antigen-induced release of eicosanoids in a type I hypersensitivity model. Guinea pigs were fed a diet containing 0.25% safflower oil (control) or 0.25% CLA [43% trans (t)10, cis (c)12; 41% c9, t11/t9, c11 18:2] for 2 wk before and during sensitization to ovalbumin (OVA). Lungs, tracheas, and bladders were incubated in physiological saline solution (PSS) for 1 h (basal mediator release) and challenged with OVA (0.01 g/l PSS) for 1 h (mediator release in response to antigen). Eicosanoids were quantified by HPLC/tandem mass spectrometry or enzyme immunoassay. CLA feeding resulted in no change in basal release but decreased eicosanoid release from sensitized tissues in response to antigen challenge in the following manner: thromboxane B(2), 6-keto-prostaglandin (PG)F(1alpha), PGF(2alpha), PGD(2), PGE(2) by 57-75% in lung, 45-65% in trachea, and 38-60% in bladder; and leukotriene C(4)/D(4)/E(4) by 87, 90, and 50% in lung, trachea, and bladder, respectively. These data indicate that feeding CLA reduces lipid-derived inflammatory mediators produced by this type I hypersensitivity model. PMID- 11893616 TI - Effects of litter size on sympathetic activity in young adult rats. AB - Rearing animals in small litters induces a permanent increase in body weight and body fat. To determine whether changes in sympathoadrenal activity contribute to this effect, litter size was adjusted the day after birth and maintained until weaning at 21 days. Sympathetic nervous system (SNS) activity was measured in adult animals using [(3)H]norepinephrine ([(3)H]NE) turnover in peripheral tissues. Although litter size was without effect on [(3)H]NE turnover in chow-fed animals, acceleration of [(3)H]NE turnover by dietary sucrose was completely abolished in heart and attenuated in interscapular brown adipose tissue and kidney of rats reared in small litters. Body and epididymal fat-pad weights were heavier in rats reared in small litters; however, weight gain in response to dietary enrichment with sucrose did not differ as a function of litter size. Thus litter size alters dietary activation of the SNS, and this effect presumably reflects changes in central nervous system regulation. PMID- 11893617 TI - Temperature dependency of force loss and Ca(2+) homeostasis in mouse EDL muscle after eccentric contractions. AB - The goals of this study were first to determine the effect of temperature on the force loss that results from eccentric contractions in mouse extensor digitorum longus (EDL) muscles and then to evaluate a potential role for altered Ca(2+) homeostasis explaining the greater isometric force loss observed at the higher temperatures. Isolated muscles performed five eccentric or five isometric contractions at either 15, 20, 25, 30, 33.5, or 37 degrees C. Isometric force loss, caffeine-induced force, lactate dehydrogenase (LDH) release, muscle accumulation of (45)Ca(2+) from the bathing medium, sarcoplasmic reticulum (SR) Ca(2+) uptake, and resting muscle fiber free cytosolic Ca(2+) concentration ([Ca(2+)](i)) were measured. The isometric force loss after eccentric contractions increased progressively as temperature rose; at 15 degrees C, there was no significant loss of force, but at 37 degrees C, there was a 30-39% loss of force. After eccentric contractions, caffeine-induced force was not affected by temperature nor was it different from that of control muscles at any temperature. Loss of cell membrane integrity and subsequent influx of extracellular Ca(2+) as indicated by LDH release and muscle (45)Ca(2+) accumulation, respectively, were minimal over the 15-25 degrees C range, but both increased as an exponential function of temperature between 30 and 37 degrees C. SR Ca(2+) uptake showed no impairment as temperature increased, and the eccentric contraction-induced rise in resting fiber [Ca(2+)](i) was unaffected by temperature over the 15-25 degrees C range. In conclusion, the isometric force loss after eccentric contractions is temperature dependent, but the temperature dependency does not appear to be readily explainable by alterations in Ca(2+) homeostasis. PMID- 11893618 TI - Acute intrarenal infusion of ANG II does not stimulate immediate early gene expression in the kidney. AB - ANG II is capable of stimulating expression of immediate early genes such as egr 1 and c-fos in a variety of cultured cells, including cells of renal origin. To investigate whether ANG II can stimulate early growth response gene expression in vivo, we studied the effects of acute renal artery infusion of low-dose ANG II (2.5 ng small middle dot kg(-1) small middle dot min(-1)) or vehicle on the renal expression of c-fos and egr-1 genes in rats. ANG II infusion for 30 or 240 min decreased renal vascular conductance by approximately 13 and 8%, respectively, compared with the vehicle group. Expression of the early growth response genes c fos and egr-1 was analyzed using Northern blot hybridization. No significant upregulation of c-fos or egr-1 mRNA levels was detected in rats that received ANG II for either 30 or 240 min, compared with the vehicle groups. We conclude that ANG II, at doses that cause significant physiological effects, does not increase the renal expression of c-fos or egr-1 genes over periods of up to 4 h in vivo. PMID- 11893619 TI - Volume expansion during acute angiotensin II receptor (AT(1)) blockade and NOS inhibition in conscious dogs. AB - The responses to AT(1)-receptor blockade (candesartan 1 mg/kg) and to concomitant volume expansion (saline 35 ml/kg for 90 min) with and without nitric oxide synthase (NOS) inhibition (N(G)-nitro-L-arginine methyl ester 30 microg small middle dot kg(-1) small middle dot min(-1)) were investigated in separate experiments in normal dogs. AT(1) blockade decreased arterial pressure (106 +/- 4 to 96 +/- 5 mmHg) and increased glomerular filtration rate (GFR) by 17% and sodium excretion threefold. NOS inhibition increased arterial pressure (103 +/- 3 to 116 +/- 3 mmHg) and decreased GFR by 21% and reduced sodium excretion by some 80%. Volume expansion increased arterial pressure significantly in all series involving this procedure, most pronounced during combined AT(1) blockade and NOS inhibition (21 +/- 4 mmHg). Volume expansion during AT(1) blockade elicited marked natriuresis (26 +/- 11 to 274 +/- 55 micromol/min) that was severely reduced by concomitant NOS inhibition (10 +/- 3 to 45 +/- 11 micromol/min), but still much larger than that seen with volume expansion during NOS inhibition alone (2 +/- 1 to 23 +/- 7 micromol/min). Volume expansion during AT(1) blockade increased GFR (+30%), less so during combined AT(1) blockade and NOS inhibition (+13%), but it did not increase GFR significantly (P = 0.07) during NOS inhibition alone. Plasma ANG II increased greater than sevenfold with AT(1) blockade and doubled with NOS inhibition (paired t-test, P < 0.05), whereas it decreased by 50-80% during volume expansion irrespective of pretreatment, i.e., during NOS inhibition, volume expansion did not generate subnormal plasma ANG II concentrations. In conclusion, 1) acute AT(1) blockade leads to hyperfiltration, natriuresis, and hyperresponsiveness to volume expansion, 2) these responses are >85% inhibitable by unspecific NOS inhibition, and 3) NOS inhibition alone is followed by increases in plasma ANG II, hypofiltration, and severe antinatriuresis that may be counterbalanced but not overwhelmed by volume expansion. Thus NOS inhibition virtually abolishes the volume expansion natriuresis, at least in part, due to the lack of appropriate inhibition of the renin-angiotensin-aldosterone system. PMID- 11893621 TI - Sympathetic control of the cardiovascular response to acute hypoxemia in the chick embryo. AB - In response to an acute hypoxemic insult, the mammalian fetus shows a redistribution of the cardiac output in favor of the heart and brain. Peripheral vasoconstriction contributes to this response and is partly mediated by the release of catecholamines. Two mechanisms of catecholamine release in the fetus are reported: 1) neurogenic sympathetic stimulation and 2) a nonneurogenic mechanism via a direct effect of hypoxemia on chromaffin tissues. In the present study, the effects of sympathetic blockade on plasma catecholamine release and cardiac output distribution in response to acute hypoxemia were studied in the chick embryo at different stages of incubation. Only at the end of the incubation period, sympathetic blockade markedly attenuated the increase in plasma catecholamine concentrations and resulted in a greater fraction of the cardiac output distributed to the carcass. However, these effects did not prevent a significant increase in cardiac output to the brain and heart during acute hypoxemia. These data imply that in the chick embryo the contribution of neurogenic mechanisms to the catecholaminergic response to acute hypoxemia becomes greater by the end of the incubation period. PMID- 11893620 TI - Volume expansion during NOS substrate donation with L-arginine: regulatory offsetting of renal response? AB - The responses to infusion of nitric oxide synthase substrate (L-arginine 3 mg.kg( 1).min(-1)) and to slow volume expansion (saline 35 ml/kg for 90 min) alone and in combination were investigated in separate experiments. L-Arginine left blood pressure and plasma ANG II unaffected but decreased heart rate (6 +/- 2 beats/min) and urine osmolality, increased glomerular filtration rate (GFR) transiently, and caused sustained increases in sodium excretion (fourfold) and urine flow (0.2 +/- 0.0 to 0.7 +/- 0.1 ml/min). Volume expansion increased arterial blood pressure (102 +/- 3 to 114 +/- 3 mmHg), elevated GFR persistently by 24%, and enhanced sodium excretion to a peak of 251 +/- 31 micromol/min, together with marked increases in urine flow, osmolar and free water clearances, whereas plasma ANG II decreased (8.1 +/- 1.7 to 1.6 +/- 0.3 pg/ml). Combined volume expansion and L-arginine infusion tended to increase arterial blood pressure and increased GFR by 31%, whereas peak sodium excretion was enhanced to 335 +/- 23 micromol/min at plasma ANG II levels of 3.0 +/- 1.1 pg/ml; urine flow and osmolar clearance were increased at constant free water clearance. In conclusion, L-arginine 1) increases sodium excretion, 2) decreases basal urine osmolality, 3) exaggerates the natriuretic response to volume expansion by an average of 50% without persistent changes in GFR, and 4) abolishes the increase in free water clearance normally occurring during volume expansion. Thus L arginine is a natriuretic substance compatible with a role of nitric oxide in sodium homeostasis, possibly by offsetting/shifting the renal response to sodium excess. PMID- 11893622 TI - IP(3)-induced tension and IP(3)-receptor expression in rat soleus muscle during postnatal development. AB - The present study was designed to examine whether changes in Ca(2+) release by inositol-1,4,5-trisphosphate (IP(3)) in 8-, 15-, and 30-day-old rat skeletal muscles could be associated with the expression of IP(3) receptors. Experiments were conducted in slow-twitch muscle in which both IP(3)-induced Ca(2+) release and IP(3)-receptor (IP(3)R) expression have been shown to be larger than in fast twitch muscle. In saponin-skinned fibers, IP(3) induced transient contractile responses in which the amplitude was dependent on the Ca(2+)-loading period with the maximal IP(3) contracture being at 20 min of loading. The IP(3) tension decreased during postnatal development, was partially inhibited by ryanodine (100 microM), and was blocked by heparin (20-400 microg/ml). Amplification of the DNA sequence encoding for IP(3)R isoforms (using the RT-PCR technique) showed that in slow-twitch muscle, the type 2 isoform is mainly expressed, and its level decreases during postnatal development in parallel with changes in IP(3) responses in immature fibers. IP(3)-induced Ca(2+) release would then have greater participation in excitation-contraction coupling in developing fibers than in mature muscle. PMID- 11893623 TI - Cholecystokinin selectively affects presympathetic vasomotor neurons and sympathetic vasomotor outflow. AB - Cholecystokinin (CCK) is a potential mediator of gastrointestinal vasodilatation during digestion. To determine whether CCK influences sympathetic vasomotor function, we examined the effect of systemic CCK administration on mean arterial blood pressure (MAP), heart rate (HR), lumbar sympathetic nerve discharge (LSND), splanchnic sympathetic nerve discharge (SSND), and the discharge of presympathetic neurons of the rostral ventrolateral medulla (RVLM) in alpha chloralose-anesthetized rats. CCK (1-8 microg/kg iv) reduced MAP, HR, and SSND and transiently increased LSND. Vagotomy abolished the effects of CCK on MAP and SSND as did the CCK-A receptor antagonist devazepide (0.5 mg/kg iv). The bradycardic effect of CCK was unaltered by vagotomy but abolished by devazepide. CCK increased superior mesenteric arterial conductance but did not alter iliac conductance. CCK inhibited a subpopulation (approximately 49%) of RVLM presympathetic neurons whereas approximately 28% of neurons tested were activated by CCK. The effects of CCK on RVLM neuronal discharge were blocked by devazepide. RVLM neurons inhibited by exogenous CCK acting via CCK-A receptors on vagal afferents may control sympathetic vasomotor outflow to the gastrointestinal tract vasculature. PMID- 11893624 TI - Glycerokinase activity in brown adipose tissue: a sympathetic regulation? AB - The effect of brown adipose tissue (BAT) sympathetic hemidenervation on the activity of glycerokinase (GyK) was investigated in different physiological conditions. In rats fed a balanced diet, the activity of the enzyme was approximately 50% lower in BAT-denervated pads than in intact, innervated pads. In rats adapted to a high-protein, carbohydrate-free diet, norepinephrine turnover rates and BAT GyK activity were already reduced, and BAT denervation resulted in a further decrease in the activity of the enzyme. Cold acclimation of normally fed rats at 4 degrees C for 10 days markedly increased the activity of the enzyme. Cold exposure (4 degrees C) for 6 h was insufficient to stimulate BAT GyK, but the activity of the enzyme was already increased after 12 h of cold exposure. The cold-induced BAT GyK stimulation was completely blocked in BAT denervated pads. The data indicate that an adequate sympathetic flow to BAT is required for the maintenance of normal levels of GyK activity and for the enzyme response to situations, such as cold exposure, which markedly increase BAT sympathetic flow. PMID- 11893625 TI - Temperature-dependent expression of sarcolemmal K(+) currents in rainbow trout atrial and ventricular myocytes. AB - Temperature has a strong influence on the excitability and the contractility of the ectothermic heart that can be alleviated in some species by temperature acclimation. The molecular mechanisms involved in the temperature-induced improvement of cardiac contractility and excitability are, however, still poorly known. The present study examines the role of sarcolemmal K(+) currents from rainbow trout (Oncorhynchus mykiss) cardiac myocytes after thermal acclimation. The two major K(+) conductances of the rainbow trout cardiac myocytes were identified as the Ba(2+)-sensitive background inward rectifier current (I(K1)) and the E-4031-sensitive delayed rectifier current (I(Kr)). In atrial cells, the density of I(K1) is very low and the density of I(Kr) is remarkably high. The opposite is true for ventricular cells. Acclimation to cold (4 degrees C) modified the two K(+) currents in opposite ways. Acclimation to cold increases the density of I(Kr) and depresses the density of I(K1). These changes in repolarizing K(+) currents alter the shape of the action potential, which is much shorter in cold-acclimated than warm-acclimated (17 degrees C) trout. These results provide the first concrete evidence that K(+) channels of trout cardiac myocytes are adaptable units that provide means to regulate cardiac excitability and contractility as a function of temperature. PMID- 11893626 TI - Electrophysiological properties of rainbow trout cardiac myocytes in serum-free primary culture. AB - A low-density primary culture of trout ventricular myocytes in serum-free growth medium was established and maintained for up to 10 days at 17 degrees C. The myocytes retained their normal rod shaped morphology, capacitive surface area of the sarcolemma (SL), and contractile quiescence. However, sarcolemmal cation currents changed significantly, some permanently, some transiently, after 8-10 days of culture. TTX-sensitive sodium current (I(Na)) and Ba(2+)-sensitive background inward rectifier potassium current (I(K1)) were permanently depressed to 24-28% of their control density measured in freshly isolated myocytes. In contrast, L-type calcium current (I(Ca)) was only transiently downregulated; after 2-3 days in culture, the density of the current was 32% of the control and recovered to the control value after 8-10 days in culture. The changes in membrane currents were reflected in the shape of the action potential (AP). After 2-3 days in culture, maximal overshoot potential and resting potential were significantly reduced, and the durations of the AP at 50 and 90% repolarization were significantly increased. These changes became significantly more pronounced after 8-10 days of culture, with the exception of AP duration at 50% repolarization level. The shortening of the early plateau phase may reflect an additional change to an outward current, presumably the rapid component of the delayed rectifier (I(Kr)). Although the present findings indicate that fish cardiac myocytes can be maintained in serum-free primary culture for at least 10 days at 17 degrees C, some but not all of the electrophysiological characteristics of the myocytes change markedly during culture. The changes in ion currents were not due to loss of sarcolemmal membrane and therefore are likely to represent altered expression of cation currents as an adaptive response to culture conditions. PMID- 11893627 TI - Brief food restriction increases FA oxidation and glycogen synthesis under insulin-stimulated conditions. AB - To determine the effects of brief food restriction on fatty acid (FA) metabolism, hindlimbs of F344/BN rats fed either ad libitum (AL) or food restricted (FR) to 60% of baseline food intake for 28 days were perfused under hyperglycemic hyperinsulinemic conditions (20 mM glucose, 1 mM palmitate, 1,000 microU/ml insulin, [3-(3)H]glucose, and [1-(14)C]palmitate). Basal glucose and insulin levels were significantly lower (P < 0.05) in FR vs. AL rats. Palmitate uptake (34.3 +/- 2.7 vs. 24.5 +/- 3.1 nmol/g/min) and oxidation (3.8 +/- 0.2 vs. 2.7 +/- 0.3 nmol.g(-1).min(-1)) were significantly higher (P < 0.05) in FR vs. AL rats, respectively. Glucose uptake was increased in FR rats and was accompanied by significant increases in red and white gastrocnemius glycogen synthesis, indicating an improvement in insulin sensitivity. Although muscle triglyceride (TG) levels were not significantly different between groups, glucose uptake and total preperfusion TG concentration were negatively correlated (r(2) = 0.27, P < 0.05). In conclusion, our results show that under hyperglycemic-hyperinsulinemic conditions, brief FR resulted in an increase in FA oxidative disposal that may contribute to the improvement in insulin sensitivity. PMID- 11893628 TI - cAMP and in vivo hypoxia induce tob, ifr1, and fos expression in erythroid cells of the chick embryo. AB - During avian embryonic development, terminal erythroid differentiation occurs in the circulation. Some of the key events, such as the induction of erythroid 2,3 bisphosphoglycerate (2,3-BPG), carbonic anhydrase (CAII), and pyrimidine 5' nucleotidase (P5N) synthesis are oxygen dependent (Baumann R, Haller EA, Schoning U, and Weber M, Dev Biol 116: 548-551, 1986; Dragon S and Baumann R, Am J Physiol Regulatory Integrative Comp Physiol 280: R870-R878, 2001; Dragon S, Carey C, Martin K, and Baumann R, J Exp Biol 202: 2787-2795, 1999; Dragon S, Glombitza S, Gotz R, and Baumann R, Am J Physiol Regulatory Integrative Comp Physiol 271: R982 R989, 1996; Dragon S, Hille R, Gotz R, and Baumann R, Blood 91: 3052-3058, 1998; Million D, Zillner P, and Baumann R, Am J Physiol Regulatory Integrative Comp Physiol 261: R1188-R1196, 1991) in an indirect way: hypoxia stimulates the release of norepinephrine (NE)/adenosine into the circulation (Dragon et al., J Exp Biol 202: 2787-2795, 1999; Dragon et al., Am J Physiol Regulatory Integrative Comp Physiol 271: R982-R989, 1996). This leads via erythroid beta adrenergic/adenosine A(2) receptor activation to a cAMP signal inducing several proteins in a transcription-dependent manner (Dragon et al., Am J Physiol Regulatory Integrative Comp Physiol 271: R982-R989, 1996; Dragon et al., Blood 91: 3052-3058, 1998; Glombitza S, Dragon S, Berghammer M, Pannermayr M, and Baumann R, Am J Physiol Regulatory Integrative Comp Physiol 271: R973-R981, 1996). To understand how the cAMP-dependent processes are initiated, we screened an erythroid cDNA library for cAMP-regulated genes. We detected three genes that were strongly upregulated (>5-fold) by cAMP in definitive and primitive red blood cells. They are homologous to the mammalian Tob, Ifr1, and Fos proteins. In addition, the genes are induced in the intact embryo during short-term hypoxia. Because the genes are regulators of proliferation and differentiation in other cell types, we suggest that cAMP might promote general differentiating processes in erythroid cells, thereby allowing adaptive modulation of the latest steps of erythroid differentiation during developmental hypoxia. PMID- 11893629 TI - Leptin secretion and hypothalamic neuropeptide and receptor gene expression in sheep. AB - Peripheral and hypothalamic mechanisms underlying the hyperphagia of lactation have been investigated in sheep. Sheep were fed ad libitum and killed at 6 and 18 days of lactation; ad libitum-fed nonlactating sheep were killed as controls. Despite increased food intake, lactating ewes were in negative energy balance. Lactation decreased plasma leptin and adipose tissue leptin mRNA concentrations. OB-Rb gene expression, determined by in situ hybridization, was increased in the hypothalamic arcuate nucleus (ARC) and ventromedial hypothalamic nucleus (VMH) at both stages of lactation. Neuropeptide Y (NPY) was increased by lactation in both the ARC and dorsomedial hypothalamus (DMH), although increased gene expression in the DMH was only apparent at day 18 of lactation. Gene expression was decreased for cocaine- and amphetamine-regulated transcript (CART) in the ARC and VMH and for proopiomelanocortin in ARC during lactation. Agouti-related peptide gene expression was increased in the ARC, and melanocortin receptor expression was unchanged in both the ARC and VMH with lactation. Thus the hypoleptinemia of lactation may activate NPY orexigenic pathways and attenuate anorexigenic melanocortin and CART pathways in the hypothalamus to promote the hyperphagia of lactation. PMID- 11893630 TI - Brain vasopressin and sodium appetite. AB - Intraventricular injections of vasopressin (VP) and antagonists with varying degrees of specificity for the VP receptors were used to identify the action of endogenous brain VP on 0.3 M NaCl intake by sodium-deficient rats. Lateral ventricular injections of 100 ng and 1 microg VP caused barrel rotations and a dramatic decrease in NaCl intake by sodium-deficient rats and suppressed sucrose intake. Intraventricular injection of the V(1)/V(2) receptor antagonist [d(CH(2))(5)(1),O-Et-Tyr(2),Val(4), Arg(8)]VP and the V(1) receptor antagonist [d(CH(2))(5)(1),O-Me-Tyr(2),Arg(8)]VP (MeT-AVP) significantly suppressed NaCl intake by sodium-deficient rats without causing motor disturbances. MeT-AVP had no effect on sucrose intake (0.1 M). In contrast, the selective V(2) receptor antagonist had no significant effect on NaCl intake. Last, injections of 100 ng MeT-AVP decreased mean arterial blood pressure (MAP), whereas 100 ng VP elevated MAP and pretreatment with MeT-AVP blocked the pressor effect of VP. These results indicate that the effects produced by 100 ng MeT-AVP represent receptor antagonistic activity. These findings suggest that the effect of exogenous VP on salt intake is secondary to motor disruptions and that endogenous brain VP neurotransmission acting at V(1) receptors plays a role in the arousal of salt appetite. PMID- 11893631 TI - L-selectin is required for fMLP- but not C5a-induced margination of neutrophils in pulmonary circulation. AB - To study the role of L-selectin in neutrophil (PMN) margination and sequestration in the pulmonary microcirculation, maximally active concentrations of C5a (900 pmol/g) and N-formylmethionyl-leucyl-phenylalanine (fMLP; 0.34 pmol/g) were injected into the jugular vein of wild-type or L-selectin-deficient C57BL/6 mice. In wild-type mice administered C5a or fMLP, 92 +/- 1% and 34 +/- 9%, respectively, of peripheral blood PMN were trapped mostly in the pulmonary circulation as determined by immunohistochemistry and myeloperoxidase activity. In wild-type mice treated with F(ab')(2) fragments of the L-selectin monoclonal antibody MEL-14 or in L-selectin-deficient mice, C5a-induced neutropenia was not significantly reduced, but the decrease in peripheral PMN in response to fMLP was completely abolished, indicating that L-selectin is necessary for fMLP- but not C5a-induced pulmonary margination. Immunostained lung sections of fMLP- or C5a treated mice showed sequestered neutrophils in alveolar capillaries with no evidence of neutrophil aggregates. We conclude that chemoattractant-induced PMN margination in the pulmonary circulation can occur by two separate mechanisms, one of which requires L-selectin. PMID- 11893632 TI - Commentary: the role of anaesthesia in interventional radiology. PMID- 11893633 TI - Review article: methodological standards in radiographer plain film reading performance studies. AB - The objectives of this paper are to raise awareness of the methodological standards that can affect the quality of radiographer plain-film reading performance studies and to determine the frequency with which these standards are fulfilled. Multiple search methods identified 30 such studies from between 1971 and the end of June 1999. The percentage of studies that fulfilled criteria for the 10 methodological standards were as follows. (1) Performance of a sample size calculation, 3%; (2) definition of a normal and abnormal report, 97%; (3) description of the sequence of events through which films passed before reporting, 94%; (4) analysis of individual groups of observers within a combination of groups, 50% (5) appropriate choice of reference standard, 80%; (6) appropriate choice of arbiter, 57%; (7) appropriate use of a control, 22%; (8) analysis of pertinent clinical subgroups, e.g. body areas, patient type, 44%; (9) availability of data for re-calculation, 59%; and (10) presentation of indeterminate results, 69%. These findings indicate variation in the application of the methodological standards to studies of radiographer's film reading performance. Careful consideration of these standards is an essential component of study quality and hence the validity of the evidence base used to underpin radiographic reporting policy. PMID- 11893634 TI - Performance of sodium iodide based (18)F-fluorodeoxyglucose positron emission tomography in the characterization of indeterminate pulmonary nodules or masses. AB - The purpose of this study was to document the accuracy of (18)F fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) with sodium iodide detectors in characterizing indeterminate lung nodules or masses and in identifying additional extra-lesional findings. 50 consecutive patients without a confident diagnosis of malignancy on CT underwent (18)FDG PET with and without attenuation correction. The diagnosis of malignancy was made using visual diagnostic criteria, and tumour-to-blood pool ratios were calculated. The final diagnosis was established by surgery, biopsy or long-term follow-up. Any additional findings made at PET were recorded and similarly verified. Using blinded visual diagnostic criteria for the differentiation of malignant from benign nodules, sodium iodide PET achieved a sensitivity of 91% (30 of 33 cases), a specificity of 88% (15 of 17 cases), a positive predictive value for malignancy of 94% (30 of 32 cases) and a negative predictive value of 83% (15 of 18 cases). False positives occurred with active tuberculosis and sarcoidosis. False negatives were a 3 cm bronchoalveolar carcinoma, a 1.3 cm sarcoma metastasis and a 1 cm carcinoma. Use of tumour-to-blood pool ratios did not improve performance. PET suggested the presence of nodal or distant metastases in 13 of 33 patients with a malignant pulmonary lesion. These PET findings were confirmed in 11 patients. These results indicate that sodium iodide PET is an accurate tool for the characterization of indeterminate pulmonary masses or nodules and simultaneously provides non-invasive staging information that can alter patient management in up to one-third of such patients. Performance of sodium iodide PET is comparable with reported results for PET scanners using other detector materials. PMID- 11893635 TI - Outcomes following unilateral uterine artery embolisation. AB - Uterine artery embolisation has been described as successful only when both arteries are embolised. However, results in patients with one congenitally absent or previously ligated artery are unknown. Women suffering from symptomatic uterine myomata were treated at a university teaching hospital, a community hospital and an outpatient surgery centre. Retrospective review of patient response to embolisation was assessed by chart review and questionnaire. Uterine and dominant fibroid size response was assessed by comparing pre- and post embolisation ultrasound examinations. This study analysed three patient groups within the general population: those who underwent unilateral embolisation because of technical failure, those who ultimately underwent bilateral embolisation after initial technical failure and those who underwent unilateral embolisation because of an absent uterine artery. 12 patients underwent unilateral embolisation, 4 of whom underwent this procedure because of an absent uterine artery. Three of these four patients had a congenitally absent uterine artery arising from the internal iliac artery and all three experienced successful outcomes. The fourth patient had a previously ligated internal iliac artery and her symptoms worsened after the procedure. Eight patients had unilateral embolisation due to technical failure. Five of these patients underwent a subsequent procedure during which the contralateral uterine artery was embolised. Four of these five patients had successful outcomes and one was lost to follow-up. Another of the eight patients suffered an arterial injury leading to technical failure, and was lost to follow-up. Of the two remaining patients with unilateral technical failure, only one had a successful outcome. This study concluded that patients who undergo unilateral embolisation for technical reasons should be offered a second embolisation procedure shortly after the initial procedure. Patients with a congenitally absent uterine artery may respond with similar success to those who underwent bilateral embolisation. In contrast, the patient with a previously ligated internal iliac artery failed. The numbers in this study are too small for statistical analysis and subsequent studies should be performed to confirm these findings. PMID- 11893636 TI - Iodixanol in paediatric gastrointestinal imaging: safety and efficacy comparison with iohexol. AB - Iodixanol (Visipaque) is a dimeric, non-ionic iodinated contrast medium that is isotonic with blood at all clinically relevant concentrations. Iodixanol was compared in a randomized, double blind, parallel group, phase III multicentre trial with a monomeric, non-ionic contrast medium, iohexol (Omnipaque), at two concentrations assessing safety, tolerability and radiographic efficacy during contrast enhanced gastrointestinal radiography examinations of children. 154 children entered the trial; 152 formed the safety population and 147 the efficacy population. All examinations were performed following standard departmental practice. Children were assigned into either a high or low concentration group (iodixanol, 150 mgI ml(-1) and 320 mgI ml(-1) vs iohexol, 140 mgI ml(-1) and 300 mgI ml(-1)). The primary outcome measure for efficacy was the overall quality of visualization, which was assessed using a 100 mm visual analogue scale (VAS). The secondary efficacy variables assessed were quality of contrast opacification, mucosal coating and overall quality of diagnostic information. Safety evaluation involved patient follow-up for at least 48 h. Taste acceptance was also assessed. There was no statistically significant difference between the two contrast media with regard to the primary and secondary efficacy variables assessed, although higher ratings were observed for iodixanol. The 100 mm VAS score overall was 86 mm for iodixanol and 82 mm for iohexol (95% confidence interval -2.56, 10.42). The frequency of adverse events was lower for patients receiving iodixanol. Adverse events, mainly diarrhoea, occurred in 12 patients (16.2%) in the iodixanol group and 28 patients (35.9%) in the iohexol group. This reached statistical significance (p=0.006). Overall, iodixanol is well suited for examinations of the gastrointestinal tract, giving good efficacy results and fewer adverse events than iohexol. PMID- 11893637 TI - Assessment of agreement between general practitioners and radiologists as to whether a radiation exposure is justified. AB - The objective of this study was to assess agreement between General Practitioners (GPs) and Consultant Radiologists as to whether a radiation exposure is justified and whether a request conforms to the Royal College of Radiologists (RCR) guidelines. Three GPs and three Consultant Radiologists were asked to review 100 requests for plain film imaging from GPs and to state whether the request justified a radiation exposure and whether the request conformed to RCR guidelines. It was discovered that there is greater agreement between radiologists than between GPs; this is a consistent pattern. The best agreement was between two Consultant Radiologists using the RCR guidelines. The poorest was between GPs using the request form details. It is suggested that the guidelines should be symptom-based to improve efficacy. PMID- 11893638 TI - A method to obtain the same levels of CT image noise for patients of various sizes, to minimize radiation dose. AB - The aim of this study was to develop a method of obtaining the same levels of CT image noise for patients of various sizes to minimize radiation dose. Two CT systems were evaluated regarding noise characteristics using phantoms and dosimetric measurements. Both CT systems performed well at dose levels used in normal clinical imaging, but only one was found to be suitable for low radiation dose applications. The CT system with the lowest noise level was used for further detailed studies. A simple strategy for manual selection of patient-specific scan parameters, considering patient size and required image quality, was implemented and verified on 11 volunteers. Images were obtained with at least the prescribed image quality at significantly reduced radiation dose levels compared with standard scan parameters. Depending on the diameter of the tomographic section, i.e. size of the subject, the dose levels could be reduced to 1-45% of the radiation dose with standard scan parameters (120 kV, 250 mAs, 10 mm). The results indicate a general potential for dose reduction in CT for slim patients. For tissue volume determination, large dose reductions can be achieved by adjusting the scan parameters for each individual. The concept of patient specific scan parameters could be fully automated in the CT system design, but would require the scan to be specified in terms of image quality rather than X ray tube load. PMID- 11893639 TI - Potential improvements in the therapeutic ratio of prostate cancer irradiation: dose escalation of pathologically identified tumour nodules using intensity modulated radiotherapy. AB - The potential of intensity modulated radiotherapy (IMRT) to improve the therapeutic ratio in prostate cancer by dose escalation of intraprostatic tumour nodules (IPTNs) was investigated using a simultaneous integrated boost technique. The prostate and organs-at-risk were outlined on CT images from six prostate cancer patients. Positions of IPTNs were transferred onto the CT images from prostate maps derived from sequential large block sections of whole prostatectomy specimens. Inverse planned IMRT dose distributions were created to irradiate the prostate to 70 Gy and all the IPTNs to 90 Gy. A second plan was produced to escalate only the dominant IPTN (DIPTN) to 90 Gy, mimicking current imaging techniques. These plans were compared with homogeneous prostate irradiation to 70 Gy using dose-volume histograms, tumour control probability (TCP) and normal tissue complication probability (NTCP) for the rectum. The mean dose to IPTNs was increased from 69.8 Gy to 89.1 Gy if all the IPTNs were dose escalated (p=0.0003). This corresponded to a mean increase in TCP of 8.7-31.2% depending on the alpha/beta ratio of prostate cancer (p<0.001), and a mean increase in rectal NTCP of 3.0% (p<0.001). If only the DIPTN was dose escalated, the TCP was increased by 6.4-27.5% (p<0.003) and the rectal NTCP was increased by 1.8% (p<0.01). In the dose escalated DIPTN IMRT plans, the highest rectal NTCP was seen in patients with IPTNs in the posterior peripheral zone close to the anterior rectal wall, and the lowest NTCP was seen with IPTNs in the lateral peripheral zone. The ratio of increased TCP to NTCP may represent an improvement in the therapeutic ratio, but was dependent on the position of the IPTN relative to the anterior rectal wall. Improvements in prostate imaging and prostate immobilization are required before clinical implementation would be possible. Clinical trials are required to confirm the clinical benefits of these improved dose distributions. PMID- 11893640 TI - Accuracy of set-up of thoracic radiotherapy: prospective analysis of 24 patients treated with radiotherapy for lung cancer. AB - In thoracic radiotherapy, a number of factors hinder the use of portal films and electronic portal imaging devices for measuring field placement errors (FPEs). The aim of this study was to assess the accuracy of treatment set-up using simulator check films (SCFs) in radiotherapy for lung cancer. Prospective evaluation was performed on 24 patients. During their radiotherapy, patients returned to the simulator weekly for a minimum of four SCFs, for which the parameters from the original simulator planning film were set, positioning being achieved without fluoroscopy. A total of 96 SCFs were taken. FPEs in left-right (L-R) and superior-inferior (S-I) direction, as well as coronal rotational errors, were measured. The mean absolute FPE was 0.35 cm in the L-R axis and 0.43 cm in the S-I axis. Statistically, the FPEs in the S-I direction were greater than those in the L-R direction (p<0.001). A margin of 0.93 cm between the clinical target volume and the planning target volume would cover 95% of FPEs in the L-R direction, whilst a margin of 1.13 cm is needed for this degree of certainty in the S-I direction. Mean coronal rotational error was 1.6 degrees. Systematic errors were greater than random errors. This study demonstrated that the FPEs were within clinical tolerance (< or = 0.7 cm) in 84.9% of the measurements. The planning margins used in our clinical practice compare favourably with the FPEs in this study. PMID- 11893641 TI - Short communication: anomalous image quality phantom scores in magnification mammography: evidence of phase contrast enhancement. AB - Anomalously high image quality scores were noted for images of the Leeds TORMAM phantom obtained using magnification mammography. Comparison of optical density profiles of fibre features in the images with non-magnified images and images previously obtained using an in-line phase contrast geometry showed the presence of phase contrast enhancement in the magnification images. The effect on the phantom score is particularly marked for this design of phantom owing to its use of fibres, which tend to enhance well. A large proportion of the phantom score is associated with fibrous features. It is concluded that direct comparison of TORMAM phantom scores from magnified images with those from non-magnified images is not valid due to the different balance of physical mechanisms forming the two kinds of image. PMID- 11893642 TI - Case report: abdominal cocoon associated with tuberculous pelvic inflammatory disease. AB - Abdominal cocoon is a rare acquired condition in which there is encapsulation of the small bowel by a fibrous membrane. The authors describe a case wherein an organism was identified for the first time. The clinical, pathological and radiological features of this unusual disease are reviewed. Peritoneal encapsulation, a related subject, is also discussed. PMID- 11893643 TI - Case report: retrocaval ureter: MR appearances. AB - We present a case of retrocaval ureter and its MR appearances. This is the first case in reported literature. The findings on i.v. urography are correlated with the MRI findings. So far CT has been the procedure of choice to confirm the diagnosis of retrocaval ureter. However, we believe MRI is likely to replace CT in the diagnosis of retrocaval ureter. PMID- 11893644 TI - Case report: Kasabach-Merritt syndrome: a review of the therapeutic options and a case report of successful treatment with radiotherapy and interferon alpha. AB - We describe the successful treatment of a neonate with Kasabach-Merritt syndrome who received local irradiation and interferon alpha therapy after failure of corticosteroid treatment. A male neonate, born after an uneventful pregnancy, had a huge haemangioma involving the upper right cervical region as well as severe thrombocytopenia. He was treated with corticosteroids, interferon alpha and radiotherapy. Prednisolone therapy (5 mg kg(-1) day(-1)) was started at 41 days of age. No therapeutic effect was observed after 2 weeks. At this time the tumour size had increased dramatically, platelet counts had decreased progressively and coagulation abnormalities had developed. Because corticosteroid therapy had been ineffective and the child was in a life-threatening condition, irradiation was delivered up to a total dose of 9.5 Gy in five fractions. Simultaneously, prednisolone therapy was slowly decreased and interferon alpha therapy (3 million U m(-2) day(-1)) was started and continued for 6 weeks. After irradiation with 9.5 Gy and beginning interferon alpha therapy, the tumour decreased in size and coagulation parameters normalized within 4 weeks. 6 months later, platelet counts and coagulation parameters were still normal. The tumour had further decreased in size. No acute severe side effects were observed. Radiation therapy combined with interferon alpha treatment is an alternative treatment modality when high dose corticoid steroid therapy has been ineffective in patients with Kasabach-Merritt syndrome, despite the risks of growth delay and secondary malignancy. In children showing no response to corticosteroids, radiotherapy and/or interferon alpha should be considered in Kasabach-Merritt syndrome. PMID- 11893645 TI - Pictorial review: adult intussusception--a CT diagnosis. AB - Intussusception, usually thought of as a childhood condition, may be encountered in adults as well, and is then more often associated with underlying pathology. While the condition is mostly unsuspected clinically, as patients present with non-specific abdominal pain that is often of long duration, CT findings are characteristic. Examples are shown of intussusception both in the small bowel and colon. Awareness of these findings allows the radiologist to make the correct diagnosis. PMID- 11893646 TI - Case of the month: congenital unilateral proptosis. PMID- 11893647 TI - Reproducibility, repeatability, correlation and measurement error. PMID- 11893649 TI - Genetic Epidemiology of COPD. PMID- 11893651 TI - Oligonucleotide microarray analysis of lung adenocarcinoma in smokers and nonsmokers identifies GPC3 as a potential lung tumor suppressor. PMID- 11893652 TI - Differential gene expression of sFRP-1 and apoptosis in pulmonary emphysema. PMID- 11893653 TI - Asthma genetics. AB - Asthma is the most common chronic childhood disease in developed nations and is a complex disease that has high social and economic costs. Asthma and its associated intermediate phenotypes are under a substantial degree of genetic control. Identifying the genes underlying asthma offers a means of better understanding its pathogenesis, with the promise of improving preventive strategies, diagnostic tools, and therapies. A number of chromosomal regions containing genes influencing asthma and atopy have been identified consistently by different groups, and a role for several candidate genes has been established. PMID- 11893654 TI - Genetic analysis of antigen-induced airway manifestations of asthma using recombinant congenic mouse strains. PMID- 11893655 TI - The genetic predisposition to interstitial lung disease: functional relevance. PMID- 11893656 TI - Sarcoidosis: association with human leukocyte antigen class II amino acid epitopes and interaction with environmental exposures. PMID- 11893657 TI - Mutations in the surfactant protein C gene associated with interstitial lung disease. PMID- 11893658 TI - Roger S. Mitchell lecture. Uses of expression microarrays in studies of pulmonary fibrosis, asthma, acute lung injury, and emphysema. AB - Expression microarrays are a powerful tool that could provide new information about the molecular pathways regulating common lung diseases. To exemplify how this tool can be useful, selected examples of informative experiments are reviewed. In studies relevant to asthma, the cytokine interleukin-13 has been shown to produce many of the phenotypic features of this disease, but the cellular targets in the airways and the molecular pathways activated are largely unknown. We have used microarrays to begin to dissect the different transcriptional responses of primary lung cells to this cytokine. In experiments designed to identify global transcriptional programs responsible for regulating lung inflammation and pulmonary fibrosis, we performed microarray experiments on lung tissue from wild-type mice and mice lacking a member of the integrin family know to be involved in activation of latent transforming growth factor (TGF) beta. In addition to identifying distinct cluster of genes involved in each of these processes, these studies led to the identification of novel pathways by which TGF-beta can regulate acute lung injury and emphysema. Together, these examples demonstrate how careful application and thorough analysis of expression microarrays can facilitate the discovery of novel molecular targets for intervening in common lung diseases. PMID- 11893660 TI - Allelic imbalance demonstrated by microsatellite analysis of lung samples from patients with familial pulmonary fibrosis. PMID- 11893659 TI - Mapping susceptibility genes for the induction of pulmonary fibrosis in mice. PMID- 11893661 TI - Overexpression of matrix metalloproteinase-7 in pulmonary fibrosis. PMID- 11893662 TI - Identification of differentially expressed genes in a monkey model of allergic asthma by microarray technology. PMID- 11893663 TI - Molecular and physiologic evidence for 5'CpG island methylation of the endothelin B receptor gene in lung cancer. PMID- 11893664 TI - Epidemiologic study of the genetics and environment of asthma, bronchial hyperresponsiveness, and atopy. PMID- 11893665 TI - Partial pneumonectomy enhances melanoma metastasis to mouse lungs. PMID- 11893666 TI - Proteomic analysis of mouse lung neoplasia. PMID- 11893667 TI - Microarray identifies cyclo-oxygenase-2-dependent modulation of insulin-like growth factor binding protein-3 in non-small cell lung cancer cells. PMID- 11893668 TI - Integrin-mediated regulation of connexin 43 expression by alveolar epithelial cells. PMID- 11893669 TI - Familial pulmonary fibrosis in the United States. PMID- 11893671 TI - Use of oligonucleotide microarrays to analyze gene expression patterns in pulmonary fibrosis reveals distinct patterns of gene expression in mice and humans. PMID- 11893670 TI - Identifying fibrosis susceptibility genes in two strains of inbred mice. PMID- 11893672 TI - Towards an effective gene therapy for idiopathic pulmonary fibrosis: anti inflammation, antifibrosis, and regeneration. PMID- 11893673 TI - Induction of cyclo-oxygenase-2 in non-small cell lung cancer cells by adenovirus vector information. PMID- 11893674 TI - Acute lung injury does not impair adenoviral-mediated gene transfer to the alveolar epithelium. PMID- 11893675 TI - Adenovirally mediated expression of urokinase receptor binding site on integrin alpha-chain blocks adhesion and migration of human lung fibroblasts. PMID- 11893676 TI - Culturing alveolar type I, alveolar type II, and mixed cell populations by altering the extracellular matrix promotes changes in intercellular signaling mechanisms. PMID- 11893677 TI - Enhancement of adenovirus-mediated gene transfer in lungs and epithelial cells by EGTA. PMID- 11893678 TI - Pseudomonas-epithelial cell interactions dissected with DNA microarrays. PMID- 11893680 TI - Thomas A. Neff lecture. Application of expression profiling to the developing lung: identification of putative regulatory networks controlling matrix production. AB - If we hope to repair damaged lung tissue associated with a variety of acquired and developmental diseases, we must first gain a full appreciation of normal lung development. As an approach, we have utilized Affymetrix (Santa Clara, CA) high density, oligonucleotide-based microarrays to generate an expression profile of the entire process of rodent lung development, which will be made publicly available. Our initial results were internally consistent and correlated closely with those generated with standard expression techniques such as Northern hybridization. We have verified known expression of genes, found other genes with previously unsuspected expression during lung development, as well as uncovered many expressed sequence tags whose role in lung development awaits further study. Data mining reveals close relationships of expression profiles between specific genes, suggesting novel regulatory relationships. In the future, application of these methods to the study of gene-targeted mice with abnormal lung development should uncover pathways of airway and alveolar development. Ultimately, expression profiling of diseased lungs might allow us to understand why the lung fails to repair, and strategies to influence repair might become apparent. PMID- 11893679 TI - Pseudomonas aeruginosa-human airway epithelial cell interaction: effects of iron on inflammation and apoptosis. PMID- 11893681 TI - Transcriptional profiling of non-small cell lung cancer using oligonucleotide microarrays. PMID- 11893682 TI - Enhancement of alveolar epithelial beta(2)-adrenergic receptor function via gene transfer. PMID- 11893683 TI - Parker B. Francis Lecture. Genetics and gene expression in pulmonary hypertension. PMID- 11893684 TI - Bone morphogenetic protein receptor 2 mutations in pulmonary hypertension. PMID- 11893686 TI - Giles F. Filley Lecture. Genetics and gene expression in lymphangioleiomyomatosis. AB - Lymphangioleiomyomatosis (LAM) is a disease of unknown etiology that is characterized by the proliferation of abnormal smooth muscle cells (LAM cells) in the lung, which leads to cystic parenchymal destruction and progressive respiratory failure. Recent evidence suggests that the proliferative and invasive nature of LAM cells may be due, in part, to somatic mutations in the TSC2 gene, which has been implicated in the pathogenesis of tuberous sclerosis complex. Here, we describe the clinical and molecular characteristics of LAM, as well as the efforts now under way to understand the genetic and biochemical factors that lead to progressive pulmonary destruction and, ultimately, to lung transplantation or death. PMID- 11893685 TI - Linkage analysis in a large family with primary pulmonary hypertension: genetic heterogeneity and a second primary pulmonary hypertension locus on 2q31-32. PMID- 11893687 TI - Microsatellite mutational analysis of endothelial cells within plexiform lesions from patients with familial, pediatric, and sporadic pulmonary hypertension. PMID- 11893688 TI - Pulmonary cysts consistent with lymphangioleiomyomatosis are common in women with tuberous sclerosis: genetic and radiographic analysis. PMID- 11893689 TI - The genetics of innate immunity. AB - Despite the tremendous interindividual variability in the response to toxins, we simply do not understand why certain people have disease develop when challenged with toxic agents, and why others remain healthy. To address this concern, we investigated whether the TLR-4 gene (toll-like receptor [TLR]4), which has been shown to affect lipopolysaccharide (LPS) responsiveness in mice, underlies the variability in airway responsiveness to inhaled LPS in humans. Here we show that common, cosegregating missense mutations (Asp299Gly and Thr399Ile) in the extracellular domain of the TLR4 receptor are associated with a significantly blunted response to inhaled LPS in 83 humans. Although in vitro findings confirm these in vivo observations, our results in humans also indicate that genes other than TLR4 may be playing a role in the biological response to LPS. To pursue this possibility, we studied genetically diverse inbred strains of mice, as well as recombinant inbred strains of mice, and have found that although TLR4 is clearly important in directing the biological response to LPS, additional genes are clearly involved in determining the physiologic and biological response to LPS in mammals. PMID- 11893690 TI - Genetic polymorphisms associated with susceptibility and outcome in ARDS. PMID- 11893691 TI - Microarray analysis indicates that pulmonary edema fluid from patients with acute lung injury mediates inflammation, mitogen gene expression, and fibroblast proliferation through bioactive interleukin-1. PMID- 11893692 TI - Acute lung injury: functional genomics and genetic susceptibility. AB - Initiated by numerous factors, acute lung injury is marked by epithelial and endothelial cell perturbation and inflammatory cell influx that leads to surfactant disruption, pulmonary edema, and atelectasis. This syndrome has been associated with a myriad of mediators including cytokines, oxidants, and growth factors. To better understand gene-environmental interactions controlling this complex process, the sensitivity of inbred mouse strains was investigated following acute lung injury that was induced by fine nickel sulfate aerosol. Measuring survival time, protein and neutrophil concentrations in BAL fluid, lung wet-to-dry weight ratio, and histology, we found that these responses varied between inbred mouse strains and that susceptibility is heritable. To assess the progression of acute lung injury, the temporal expression of genes and expressed sequence tags was assessed by complementary DNA microarray analysis. Enhanced expression was noted in genes that were associated with oxidative stress, antiprotease function, and extracellular matrix repair. In contrast, expression levels of surfactant proteins (SPs) and Clara cell secretory protein (ie, transcripts that are constitutively expressed in the lung) decreased markedly. Genome-wide analysis was performed with offspring derived from a sensitive and resistant strain (C57BL/6xA F(1) backcrossed with susceptible A strain). Significant linkage was identified for a locus on chromosome 6 (proposed as Aliq4), a region that we had identified previously following ozone-induced acute lung injury. Two suggestive linkages were identified on chromosomes 1 and 12. Using haplotype analysis to estimate the combined effect of these regions (along with putative modifying loci on chromosomes 9 and 16), we found that five loci interact to account for the differences in survival time of the parental strains. Candidate genes contained in Aliq4 include SP-B, aquaporin 1, and transforming growth factor-alpha. Thus, the functional genomic approaches of large gene set expression (complementary DNA microarray) and genome-wide analyses continue to provide novel insights into the genetic susceptibility of lung injury. PMID- 11893693 TI - Lipopolysaccharide stimulation of the human neutrophil: an analysis of changes in gene transcription and protein expression by oligonucleotide microarrays and proteomics. PMID- 11893694 TI - Lipopolysaccharide-induced neutrophil gene expression under in vivo or in vitro conditions. PMID- 11893695 TI - Role of keratinocyte growth factor in regulating lipogenesis in alveolar type II cells: a gene-profiling approach. PMID- 11893696 TI - Up-regulation of angiogenic factor expression in hypoxia-treated mouse lung demonstrated by DNA array technique. PMID- 11893697 TI - Identification of novel target proteins in Th2-dominated allergic inflammatory responses using complementary DNA representational difference analysis and complementary DNA microarrays. PMID- 11893699 TI - Secretion-competent mouse tracheal epithelial cell culture from the genetically altered mouse: pathway analysis via gene array. PMID- 11893698 TI - Subtractive hybridization analysis of Pneumocystis carinii gene activation induced by interaction with lung epithelial cells and matrix. PMID- 11893700 TI - Use of spotted oligonucleotide arrays for large-scale analysis of mammalian gene expression. PMID- 11893701 TI - Gene expression in an in vitro model of epithelial repair. PMID- 11893702 TI - Genetic and environmental risk factors in beryllium sensitization and chronic beryllium disease. PMID- 11893703 TI - Microarray analysis of gene expression in the embryonic lung. PMID- 11893704 TI - Quantitative trait loci that regulate susceptibility to both butylated hydroxytoluene-induced pulmonary inflammation and lung tumor promotion in CXB recombinant inbred mice. PMID- 11893705 TI - Primary pulmonary hypertension is predominantly a hereditary disease. PMID- 11893706 TI - Hypoxia up-regulates mouse vascular endothelial growth factor D promoter activity in rat pulmonary microvascular smooth-muscle cells. PMID- 11893707 TI - Specific bone morphogenic protein receptor II mutations found in primary pulmonary hypertension cause different biochemical phenotypes in vitro. PMID- 11893708 TI - Pulmonary gene expression profiles of spontaneously hypertensive rats exposed to environmental tobacco smoke. PMID- 11893709 TI - Interleukin-18 expression in cystic fibrosis lungs. PMID- 11893710 TI - Intracellular adhesion molecule Gly241Arg polymorphism has no impact on ARDS or septic shock in community-acquired pneumonia. PMID- 11893711 TI - Impact of cytokine gene polymorphisms on outcomes of coronary artery bypass graft surgery. PMID- 11893712 TI - Tumor necrosis factor gene polymorphisms and the variable presentation and outcome of community-acquired pneumonia. PMID- 11893713 TI - Roles of interleukin-13 and interferon-gamma in lung inflammation. PMID- 11893714 TI - High and low inflammatory response phenotypes in 101 normal human subjects. PMID- 11893715 TI - Impaired clearance of apoptotic cells from cystic fibrosis airways. PMID- 11893716 TI - A search for linkage to atopic asthma in candidate regions in a Danish population. PMID- 11893718 TI - Regulation of vascular smooth muscle cell growth and adhesion by paired-related homeobox genes. PMID- 11893717 TI - Application of expression microarrays to the investigation of fetal lung development in a glucocorticoid receptor knockout mouse model. PMID- 11893719 TI - Gene therapy of mucus hypersecretion in experimental asthma. PMID- 11893720 TI - Mutations in DNAI1 (IC78) cause primary ciliary dyskinesia. PMID- 11893721 TI - Mode of action of RNA/DNA oligonucleotides: progress in the development of gene repair as a therapy for alpha(1)-antitrypsin deficiency. AB - We describe a technology developed for the site-specific correction of a single base carried on an episome or chromosome in prokaryotic and eukaryotic cells. Critical to the development of this technology as a therapeutic device for treating genetic disorders, like alpha(1)-antitrypsin deficiency, is the establishment of a standardized assay to study its mode of action and structure activity relationships (SARs). To this end, a positive-selection system in Escherichia coli has been developed to assess RNA/DNA oligonucleotide (RDO) directed repair activity. We demonstrate that RDO-directed repair requires the concerted action of the two following repair proteins: the pairing protein RecA; and the mismatch recognition protein, MutS. SAR studies demonstrate that the RDO molecule is functionally asymmetric. The RNA-containing strand enables strand pairing and stabilization of the molecule, and the DNA-containing strand confers the information transfer. PMID- 11893722 TI - Hyperplasia of pulmonary artery smooth muscle cells is causally related to overexpression of the serotonin transporter in primary pulmonary hypertension. PMID- 11893723 TI - Recombinant adeno-associated virus gene therapy for cystic fibrosis and alpha(1) antitrypsin deficiency. PMID- 11893725 TI - Gene expression profiles in pulmonary hypertension. PMID- 11893724 TI - Cyclooxygenase-2 overexpression, using an integrin-targeted gene delivery system (the LID vector), inhibits fibroblast proliferation in vitro and leads to increased prostaglandin E(2) in the lung. PMID- 11893726 TI - Pulmonary genetics, genomics, and gene therapy: conference summary. PMID- 11893728 TI - Reciprocal recruitment of DRIP/mediator and p160 coactivator complexes in vivo by estrogen receptor. AB - Two functionally distinct classes of coactivators are recruited by liganded estrogen receptor, the DRIP/Mediator complex and p160 proteins, although the relative dynamics of recruitment is unclear. Previously, we have shown a direct, estradiol-dependent interaction between the DRIP205 subunit of the DRIP complex and the estrogen receptor (ER) AF2 domain. Here we demonstrate the in vivo recruitment of other endogenous DRIP subunits to ER in response to estradiol treatment in MCF-7 cells. To explore the relationship between DRIP and p160 coactivators, we examined the kinetics of coactivator recruitment to the ER target promoter, pS2, by chromatin immunoprecipitation. We observed a cyclic association and dissociation of coactivators with the promoter, with recruitment of p160s and DRIPs occurring in opposite phases, suggesting an exchange between these coactivator complexes at the target promoter. PMID- 11893729 TI - Androgen receptor-interacting protein 3 and other PIAS proteins cooperate with glucocorticoid receptor-interacting protein 1 in steroid receptor-dependent signaling. AB - Androgen receptor (AR)-interacting protein 3 (ARIP3/PIASxalpha) is a coregulator capable of modulating transcriptional activity of various steroid receptors. We have characterized functional regions of ARIP3 and studied its interaction with the glucocorticoid receptor (GR)-interacting protein 1 (GRIP1). We find that the potential zinc-binding domain is critical for ARIP3 to function as a coactivator; the deletion of amino acids 347-418 or the mutation of the conserved cysteines 385 and 388 to serines converts ARIP3 to a transcriptional repressor from AR dependent minimal promoters and abolishes its ability to activate GR. By contrast, mutations in the two LXXLL motifs of ARIP3 have relatively minor effects on its ability to regulate AR or GR function. ARIP3 is able to interact with different regions of GRIP1, but the strongest interaction is detected with the C-terminal region (amino acids 1122-1462) of GRIP1. The interaction of ARIP3 with the latter GRIP1 domain or full-length GRIP1 and the ability of ARIP3 to cooperate with GRIP1 in the regulation of AR- or GR-dependent transcription are dependent on the ARIP3 zinc-binding region. We also find a strong synergism between GRIP1 and two other PIAS family members, Miz1 and PIAS1. Taken together, our results suggest that PIAS proteins and GRIP1 interact functionally in transcriptional regulation. PMID- 11893730 TI - Heterogeneous nuclear ribonucleoprotein A1 interferes with the binding of the human T cell leukemia virus type 1 rex regulatory protein to its response element. AB - The human T cell leukemia virus, type 1 (HTLV-1), Rex protein mediates the nuclear export of unspliced and incompletely spliced viral mRNAs. This post transcriptional activity is dependent in part on the binding of this protein to cis-regulatory sequences termed the Rex-response element (XRE). We have proposed previously that the decreased functionality exhibited by Rex in human lymphoblastoid Jurkat T cells may be linked to alterations in the Rex/XRE interactions. The analysis of the ribonucleoprotein complexes formed between Jurkat nuclear proteins and XRE-RNA led to the identification of a 36-kDa protein as heterogeneous nuclear ribonucleoprotein (hnRNP) A1. In vitro binding assays revealed that hnRNP A1 proteins were found to interfere with the binding of Rex to XRE, whereas nuclear extracts depleted of these proteins were unable to disrupt Rex-XRE complexes. Furthermore, A1 proteins from Jurkat cells were acting in a concentration-dependent manner, suggesting that the amount of these RNA binding proteins is a critical parameter in controlling Rex activity. We indeed observed a lower level of hnRNP A1 in in vitro HTLV-1-transformed virus-producing T cells than that detected in Jurkat cells. Likewise, overexpression of hnRNP A1 proteins in 293T cells and in Jurkat cells led to a decrease in the expression of a reporter gene dependent on Rex/XRE interactions. Such a decrease was not observed when the expression of the same reporter gene by cells overexpressing hnRNP A1 was dependent on the interactions of human immunodeficiency virus Rev protein with the Rev-response element. These findings indicate that hnRNP A1 by competing with Rex for the formation of REX-XRE complexes is specifically involved in the modulation of the post-transcriptional activity of Rex. PMID- 11893731 TI - Interleukin-13 gene expression is regulated by GATA-3 in T cells: role of a critical association of a GATA and two GATG motifs. AB - Using a transgenic approach, we studied the role of GATA-3 in T cells. As previously shown, enforced GATA-3 expression in transgenic mice inhibits Th1 differentiation of CD4 T cells, but unexpectedly, both type 1 (interferon gamma) and type 2 (interleukin (IL)-4 and IL-13) cytokine genes were activated in the transgenic CD8 T cells. Because IL-13 gene expression was highly enhanced in vivo by GATA-3 expression, we studied the human and the mouse IL-13 gene promoters and found an evolutionary-conserved association of a consensus GATA binding site and two GATG motifs. We showed that efficient GATA-3 binding to this regulatory sequence required these three motifs and that the affinity of the GATA zinc fingers for this association was five times higher than for the consensus GATA binding site alone. Transfections in a T cell line or transactivation by GATA-3 showed that the combination of the three sites was required for full transcriptional activity of the IL-13 gene promoter. Finally we showed that this association of binding sites causes a very high sensitivity of the IL-13 gene promoter to small variations in the level of GATA-3 protein. Altogether, these results indicate an important role of GATA-3 in CD8 cytokine gene expression and demonstrate that a critical network of GATA binding sites highly modulates GATA-3 activity. PMID- 11893732 TI - Intracellular localization and preassembly of the NADPH oxidase complex in cultured endothelial cells. AB - The phagocyte-type NADPH oxidase expressed in endothelial cells differs from the neutrophil enzyme in that it exhibits low level activity even in the absence of agonist stimulation, and it generates intracellular reactive oxygen species. The mechanisms underlying these differences are unknown. We studied the subcellular location of (a) oxidase subunits and (b) functionally active enzyme in unstimulated endothelial cells. Confocal microscopy revealed co-localization of the major oxidase subunits, i.e. gp91(phox), p22(phox), p47(phox), and p67(phox), in a mainly perinuclear distribution. Plasma membrane biotinylation experiments confirmed the predominantly (>90%) intracellular distribution of gp91(phox) and p22(phox). After subcellular protein fractionation, approximately 50% of the gp91(phox) (91-kDa band), p22(phox), p67(phox), and p40(phox) pools and approximately 30% of the p47(phox) were present in the 1475 x g ("nucleus-rich") fraction. Likewise, approximately 50% of total NADPH-dependent O(2)() production (assessed by lucigenin (5 microm) chemiluminescence) was found in the 1475 x g fraction. Co-immunoprecipitation studies and measurement of NADPH-dependent reactive oxygen species production (cytochrome c reduction assay) demonstrated that p22(phox), gp91(phox), p47(phox), p67(phox), and p40(phox) existed as a functional complex in the cytoskeletal fraction. These results indicate that, in contrast to the neutrophil enzyme, a substantial proportion of the NADPH oxidase in unstimulated endothelial cells exists as a preassembled intracellular complex associated with the cytoskeleton. PMID- 11893734 TI - Biochemical characterization of the core structure of alpha-synuclein filaments. AB - Intracellular filamentous aggregates comprised of alpha-synuclein such as Lewy bodies and glial cytoplasmic inclusions are the defining hallmarks of a subset of neurodegenerative diseases including Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. We have analyzed biochemical and structural properties of alpha-synuclein filaments assembled in vitro or extracted from brains of patients with multiple system atrophy and found that both types of filaments are insoluble to detergents and partially resistant to proteinase K digestion. Immunoelectron microscopy and immunoblot analysis showed that both amino and carboxyl termini of alpha-synuclein in in vitro assembled filaments were degraded by proteinase K treatment, whereas the central portion of alpha synuclein is resistant to proteinase K and retains filamentous structures. Protein sequencing and mass spectrometric analyses of the proteinase K-resistant, minimal fragment of 7 kDa revealed that amino acid residues 31-109 of alpha synuclein constitute the core unit of the filaments. These observations suggest that the central half of the alpha-synuclein polypeptide, containing five tandem repeats as well as a part of the carboxyl-terminal acidic region, forms the core structure of alpha-synuclein filaments, which is coated by the amino- and carboxyl-terminal portions at the periphery. PMID- 11893733 TI - Activation of the murine type II transforming growth factor-beta receptor gene: up-regulation and function of the transcription factor Elf-3/Ert/Esx/Ese-1. AB - Previous studies demonstrated that differentiation of mouse embryonal carcinoma cells leads to transcriptional up-regulation of the mouse type II transforming growth factor-beta receptor (mTbetaR-II) gene. To elucidate the molecular mechanisms regulating transcription of this gene, we isolated the 5'-flanking region of the mTbetaR-II gene and characterized its expression in F9 differentiated cells. Analysis of mTbetaR-II promoter/reporter gene constructs demonstrates that two conserved Ets-binding sites play an important role in the activity of the mTbetaR-II promoter. Importantly, we present evidence that mElf 3, a member of the Ets family, plays a key role in the activation of the mTbetaR II promoter. Northern blot analysis reveals that the steady-state levels of mTbetaR-II mRNA increase in parallel with those of mElf-3 mRNA during the differentiation of F9 embryonal carcinoma cells. We also demonstrate that mElf-3 contains one or more domains that influence its binding to DNA. Finally, we report that a single amino acid substitution in the transactivation domain of mElf-3 reduces its ability to transactivate and elevates its steady-state levels of expression. In conclusion, our data argue that mElf-3 plays a key role in the regulation of the mTbetaR-II gene, and Elf-3 itself is regulated at multiple levels. PMID- 11893735 TI - Metabolic channeling of carbamoyl phosphate, a thermolabile intermediate: evidence for physical interaction between carbamate kinase-like carbamoyl phosphate synthetase and ornithine carbamoyltransferase from the hyperthermophile Pyrococcus furiosus. AB - Two different approaches provided evidence for a physical interaction between the carbamate kinase-like carbamoyl-phosphate synthetase (CKase) and ornithine carbamoyltransferase (OTCase) from the hyperthermophilic archaeon Pyrococcus furiosus. Affinity electrophoresis indicated that CKase and OTCase associate into a multienzyme cluster. Further evidence for a biologically significant interaction between CKase and OTCase was obtained by co-immunoprecipitation combined with formaldehyde cross-linking experiments. These experiments support the hypothesis that CKase and OTCase form an efficient channeling cluster for carbamoyl phosphate, an extremely thermolabile and potentially toxic metabolic intermediate. Therefore, by physically interacting with each other, CKase and OTCase prevent the thermodenaturation of carbamoyl phosphate in the aqueous cytoplasmic environment. PMID- 11893736 TI - Importance of the histidine ligand to coenzyme B12 in the reaction catalyzed by methylmalonyl-CoA mutase. AB - Methylmalonyl-CoA mutase is an adenosylcobalamin (AdoCbl)-dependent enzyme that catalyzes the rearrangement of methylmalonyl-CoA to succinyl-CoA. The crystal structure of this protein revealed that binding of the cofactor is accompanied by a significant conformational change in which dimethylbenzimidazole, the lower axial ligand to the cobalt in solution, is replaced by His-610 donated by the active site. The contribution of the lower axial base to the approximately 10(12) fold rate acceleration of the homolytic cleavage of the upper axial cobalt-carbon bond has been the subject of intense scrutiny in the model inorganic literature. In contrast, trans ligand effects in methylmalonyl-CoA mutase and indeed the significance of the ligand replacement are poorly understood. In this study, we have used site-directed mutagenesis to create the H610A and H610N variants of methylmalonyl-CoA mutase and report that both mutations exhibit both diminished activity (5,000- and 40,000-fold, respectively) and profoundly weakened affinity for the native cofactor, AdoCbl. In contrast, binding of the truncated cofactor analog, adenosylcobinamide, lacking the nucleotide tail, is less impaired. The catalytic failure of the His-610 mutants is in marked contrast to the phenotype of the adenosylcobinamide-GDP reconstituted wild type enzyme that exhibits only a 4-fold decrease in activity, although His-610 fails to coordinate when this cofactor analog is bound. Together, these studies suggest that His-610 may: (i) play a structural role in organizing a high affinity cofactor binding site possibly via electrostatic interactions with Asp-608 and Lys-604, as suggested by the crystal structure and (ii) play a role in catalyzing the displacement of dimethylbenzimidazole thereby facilitating the conformational change that must precede cofactor docking to the mutase active site. PMID- 11893737 TI - Specific modulation of Kex2/furin family proteases by potassium. AB - Kex2 protease is the prototype for a family of proteases responsible for endoproteolytic cleavage at multi-basic motifs in the eukaryotic secretory pathway. Here we demonstrate that potassium ion can act as a modulator of Kex2 activity with an apparent affinity of approximately 20 mm. Other monovalent cations (Li(+), Na(+), etc.) display similar effects, but affinities are all over 20-fold lower. Potassium ion binding stimulates turnover at physiologically relevant Lys-Arg cleavage sites but reduces turnover with at least one incorrect sequence. Furthermore, the mammalian Kex2 homolog furin displays similar effects. In contrast, the neuroendocrine homolog PC2 is inhibited by potassium ion with all substrates examined. The pre-steady-state behavior of Kex2 is also altered upon binding of potassium ion, with opposite effects on acylation and deacylation rates. These biochemical data indicate that potassium ion concentration may function as a regulator of processing protease specificity and activity in the eukaryotic secretory pathway, with such enzymes potentially encountering compartments high in potassium ion caused by the action of antiporters such as yeast NHX1 (VPS44) or the mammalian NHE7. PMID- 11893738 TI - Histone acetylation in vivo at the osteocalcin locus is functionally linked to vitamin D-dependent, bone tissue-specific transcription. AB - The accessibility of regulatory elements in chromatin represents a principal rate limiting parameter of gene transcription and is modulated by enzymatic transcriptional co-factors that alter the topology of chromatin or covalently modify histones (e.g. by acetylation). The bone-specific activation and 1,25 dihydroxyvitamin D(3) enhancement of osteocalcin (OC) gene transcription are both functionally linked to modifications in nucleosomal organization. The initiation of tissue-specific basal transcription is accompanied by the induction of two DNase I hypersensitive sites, and this chromatin remodeling event requires binding of the key osteogenic factor RUNX2/CBFA1 to the OC promoter. Here, we analyzed the acetylation status of histones H3 and H4 when the OC gene is active (in osteoblastic ROS17/2.8 cells) or inactive (in fibroblastic ROS24/1 cells) using chromatin immunoprecipitation assays. We find that acetylated histone H3 and H4 proteins are associated with the OC promoter only when the gene is transcriptionally active and that the acetylation status is relatively uniform across the OC locus under basal conditions. Acetylation of H4 at the OC gene is selectively increased following vitamin D(3) enhancement of OC transcription, with the most prominent changes occurring in the region between the vitamin D(3) enhancer and basal promoter. Thus, our results suggest functional linkage of H3 and H4 acetylation in specific regions of the OC promoter to chromatin remodeling that accompanies tissue-specific transcriptional activation and vitamin D enhancement of OC gene expression. These findings provide mechanistic insights into bone-specific gene activation within a native genomic context in response to steroid hormone-related regulatory cues. PMID- 11893739 TI - RANTES-mediated chemokine transcription in astrocytes involves activation and translocation of p90 ribosomal S6 protein kinase (RSK). AB - RANTES (regulated on activation normal T cell expressed and secreted) (> or =10 ng/ml) stimulates the induction of KC and other chemokines in astrocytes. Elements of the signal transduction pathway controlling this response were identified. RANTES induced phosphorylation of MEK, ERK1/2, p90 ribosomal S6 kinases (RSK), and cAMP-response element-binding protein (CREB) in astrocytes. U0126, a pharmacological inhibitor of MEK, blocked the phosphorylation of the downstream elements ERK, RSK, and CREB, inhibited chemokine synthesis, and reduced transcription from a KC promoter construct. Dominant negative mutants of RSK or CREB blocked the transcription driven by the KC promoter. Finally, RANTES treatment induces nuclear translocation of phosphorylated RSK in astrocytes. This novel role for RSK in signaling chemokine responses and synthesis in astrocytes may contribute to the amplification mechanisms responsible for prolonging inflammatory responses in the central nervous system. PMID- 11893740 TI - Interactions between fission yeast mRNA capping enzymes and elongation factor Spt5. AB - Elongating RNA polymerase II is targeted by macromolecular assemblies that regulate mRNA synthesis and processing. The capping apparatus is the first of the assemblies to act on the nascent pre-mRNA. Although recruitment of the capping enzymes to the transcription complex is dependent on phosphorylation of the C terminal domain of the Rpb1 subunit of polymerase II (Pol-II), there may be additional levels of control that coordinate capping with elongation. Here we show that the triphosphatase (Pct1) and guanylyltransferase (Pce1) enzymes of the fission yeast capping apparatus bind independently to the elongation factor Spt5. The C-terminal domain of the 990-amino acid Schizosaccharomyces pombe Spt5 protein, composed of repeats of a nonapeptide motif (consensus sequence TPAWNSGSK), is necessary and sufficient for binding to the capping enzymes in vivo (in a two-hybrid assay) and in vitro. As few as four nonamer repeats suffice for Spt5 binding to Pct1 in vitro, whereas six repeats are required for Spt5 binding to Pce1. A 116-amino acid fragment of the guanylyltransferase Pce1 suffices for binding to the Spt5 C-terminal domain (CTD) but not for binding to the Pol-II CTD. Pct1 and Pce1 can bind simultaneously to the Spt5 CTD in vitro. We find that Spt5 is essential for viability of S. pombe and that it interacts in vivo with S. pombe Spt4 via a central domain distinct from the Spt5 CTD. We suggest that Spt5-induced arrest of elongation at promoter proximal positions ensures a temporal window for recruitment of the capping enzymes. PMID- 11893741 TI - The unfolding pathway of leech carboxypeptidase inhibitor. AB - The unfolding and denaturation curves of leech carboxypeptidase inhibitor (LCI) were elucidated using the technique of disulfide scrambling. In the presence of thiol initiator and denaturant, the native LCI denatures by shuffling its native disulfide bonds and transforms into a mixture of scrambled species. 9 of 104 possible scrambled isomers of LCI, amounting to 90% of total denatured LCI, can be distinguished. The denaturation curve that plots the fraction of native LCI converted into scrambled isomers upon increasing concentrations of denaturant shows that the concentration of guanidine thiocyanate and guanidine hydrochloride required to reach 50% of denaturation is 2.4 and 3.6 m, respectively. In contrast, native LCI is resistant to urea denaturation even at high concentration (8 m). The LCI unfolding pathway was defined based on the evolution of the relative concentration of scrambled isoforms of LCI upon denaturation. Two populations of scrambled species suffer variations along the unfolding pathway. One accumulates as intermediates under strong denaturing conditions and corresponds to open or relaxed structures, among which the beads-form isomer is found. The other population shows an inverse correlation between their relative abundances and the denaturing conditions and should have another kind of non native structure that is more compact than the unfolded state. The rate constants of unfolding of LCI are low when compared with other disulfide-containing proteins. Overall, the results presented in this study show that LCI, a molecule with potential biotechnological applications, has slow kinetics of unfolding and is highly stable. PMID- 11893742 TI - Functional up-regulation of HERG K+ channels in neoplastic hematopoietic cells. AB - Kv1.3 channels regulate proliferation of normal lymphocytes, but the role of voltage-gated potassium channels in transformed hematopoietic cells is not known. We examined transcripts for Kv1.3, h-erg, h-eag, and BEC1 genes in primary lymphocytes and leukemias and in several hematopoietic cell lines. Surprisingly, BEC1, formerly thought to be brain-specific, was present in all the primary leukemias examined, in resting peripheral blood lymphocytes, and in proliferating activated tonsillar cells, lymphocytes from Sjogren's patients, and Epstein-Barr virus-transformed B-cells. Only h-erg mRNA was up-regulated in the cancer cells, but this was not due to proliferation per se, because it was not elevated in any of the proliferating noncancerous lymphocyte types examined. Nor did h-erg transcript levels correlate with the B-cell subset, because it was elevated in immature neoplastic B-CLL cells (CD5(+)) and in a CD5(-) Burkitt's lymphoma cell line (Raji) but not in Sjogren's syndrome cells (enriched in CD5(+) B-cells) or Epstein-Barr virus-transformed B-cells, which are mature CD5(-) B-cells. The protein and whole cell current levels roughly corresponded with the amount of mRNA expressed in three hematopoietic cell lines: CEM (an acute lymphoblastic leukemic line), K562 (a chronic myelogenous leukemic line), and U937 (an acute promyelocytic leukemic line). The selective HERG channel blocker, E-4031, reduced proliferation of CEM, U937, and K562 cells, and this appears to be the first direct evidence of a functional role for the HERG current in cancer cells. Selective up-regulation of h-erg appears to occur in neoplastic hematopoietic cells, thus providing a marker and potential therapeutic target. PMID- 11893743 TI - Structural identification of 2'- and 3'-O-acetyl-ADP-ribose as novel metabolites derived from the Sir2 family of beta -NAD+-dependent histone/protein deacetylases. AB - The Sir2 (silent information regulator 2) family of histone/protein deacetylases has been implicated in a wide range of biological activities, including gene silencing, life-span extension, and chromosomal stability. Their dependence on beta-NAD(+) for activity is unique among the known classes of histone/protein deacetylase. Sir2 enzymes have been shown to couple substrate deacetylation and beta-NAD(+) cleavage to the formation of O-acetyl-ADP-ribose, a newly described metabolite. To gain a better understanding of the catalytic mechanism and of the biological implications of producing this molecule, we have performed a detailed enzymatic and structural characterization of O-acetyl-ADP-ribose. Through the use of mass spectrometry, rapid quenching techniques, and NMR structural analyses, 2' O-acetyl-ADP-ribose and 3'-O-acetyl-ADP-ribose were found to be the solution products produced by the Sir2 family of enzymes. Rapid quenching approaches under single-turnover conditions identified 2'-O-acetyl-ADP-ribose as the enzymatic product, whereas 3'-O-acetyl-ADP-ribose was formed by intramolecular transesterification after enzymatic release into bulk solvent, where 2'- and 3'-O acetyl-ADP-ribose exist in equilibrium (48:52). In addition to (1)H and (13)C chemical shift assignments for each regioisomer, heteronuclear multiple-bond correlation spectroscopy was used to assign unambiguously the position of the acetyl group. These findings are highly significant, because they differ from the previous conclusion, which suggested that 1'-O-acetyl-ADP-ribose was the solution product of the reaction. Possible mechanisms for the generation of 2'-O-acetyl ADP-ribose are discussed. PMID- 11893744 TI - Cyclic AMP-induced forkhead transcription factor, FKHR, cooperates with CCAAT/enhancer-binding protein beta in differentiating human endometrial stromal cells. AB - Decidual transformation of human endometrial stromal (ES) cells requires sustained activation of the protein kinase A (PKA) pathway. In a search for novel transcriptional mediators of this process, we used differential display PCR analysis of undifferentiated primary ES cells and cells stimulated with 8-bromo cAMP (8-Br-cAMP). We now report on the role of forkhead homologue in rhabdomyosarcoma (FKHR), a recently described member of the forkhead/winged-helix transcription factor family, as a mediator of endometrial differentiation. Sustained 8-Br-cAMP stimulation resulted in the induction and nuclear accumulation of FKHR in differentiating ES cells. Immunohistochemical studies revealed that endometrial stromal expression of FKHR in vivo is confined to decidualizing cells during the late secretory phase of the cycle and coincides with the expression of CCAAT/enhancer-binding protein beta (C/EBPbeta). Reporter gene studies showed that FKHR potently enhances PKA-dependent activation of the tissue-specific decidual prolactin (dPRL) promoter, a major differentiation marker in human ES cells. Transcriptional augmentation by FKHR was effected through functional cooperation with C/EBPbeta and binding to a composite FKHR C/EBPbeta response unit in the proximal promoter region. Furthermore, FKHR and C/EBPbeta were shown to interact directly in a glutathione S-transferase pull down assay. These results provide the first evidence of regulated expression of FKHR and demonstrate that FKHR has an integral role in PKA-dependent endometrial differentiation through its ability to bind and functionally cooperate with C/EBPbeta. PMID- 11893745 TI - Interaction of the eukaryotic elongation factor 1A with newly synthesized polypeptides. AB - eEF1A, the eukaryotic homologue of bacterial elongation factor Tu, is a well characterized translation elongation factor responsible for delivering aminoacyl tRNAs to the A-site at the ribosome. Here we show for the first time that eEF1A also associates with the nascent chain distal to the peptidyltransferase center. This is demonstrated for a variety of nascent chains of different lengths and sequences. Interestingly, unlike other ribosome-associated factors, eEF1A also interacts with polypeptides after their release from the ribosome. We demonstrate that eEF1A does not bind to correctly folded full-length proteins but interacts specifically with proteins that are unable to fold correctly in a cytosolic environment. This association was demonstrated both by photo-cross-linking and by a functional refolding assay. PMID- 11893746 TI - Nuclear localization signal of murine CMP-Neu5Ac synthetase includes residues required for both nuclear targeting and enzymatic activity. AB - 5-N-Acetylneuraminic acid (Neu5Ac) is the major sialic acid derivative found in animal cells. As a component of cell surface glycoconjugates, Neu5Ac is pivotal to numerous cellular recognition and communication processes including host parasite interactions. A prerequisite for the synthesis of sialylated glycoconjugates is the activation of Neu5Ac to cytidine-monophosphate N acetylneuraminic acid (CMP-Neu5Ac). The reaction is catalyzed by CMP-Neu5Ac synthetase (syn), which, for unknown reasons, resides in the nucleus. Sequence analysis of the cloned murine CMP-Neu5Ac synthetase identified three clusters of basic amino acids (BC1-BC3) that might function as nuclear localization signals (NLS). In the present study chimeric protein and mutagenesis strategies were used to show that BC1 and BC2 are active NLS sequences when attached to the green fluorescent protein (enhanced GFP), but only BC2 is necessary and sufficient to mediate the nuclear import of CMP-Neu5Ac synthetase. Site-directed mutations identified the residues K(198)RXR to be essential for nuclear transport and Arg(202) to be necessary to complete the transport process. Cytoplasmic forms of CMP-Neu5Ac synthetase generated by single site mutations in BC2 demonstrated that (i) enzyme activity is independent of nuclear localization, and (ii) Arg(199) and Arg(202) are involved in both nuclear transport and synthetase activity. Comparison of all known and predicted CMP-sialic acid synthetases reveals Arg(202) and Gln(203) as highly conserved in evolution and critically important for optimal synthetase activity but not for nuclear localization. Combined, the data demonstrate that nuclear transport and enzyme activity are independent functions that share some common amino acid requirements in CMP-Neu5Ac synthetase. PMID- 11893747 TI - Environment and mobility of a series of fluorescent reporters at the amino terminus of structurally related peptide agonists and antagonists bound to the cholecystokinin receptor. AB - Fluorescence is a powerful biophysical tool for the analysis of the structure and dynamics of proteins. Here, we have developed two series of new fluorescent probes of the cholecystokinin (CCK) receptor, representing structurally related peptide agonists and antagonists. Each ligand had one of three distinct fluorophores (Alexa(488), nitrobenzoxadiazolyl, or acrylodan) incorporated in analogous positions at the amino terminus just outside the hormone's pharmacophore. All of the probes bound to the CCK receptor specifically and with high affinity, and intracellular calcium signaling studies showed the chemically modified peptides to be fully biologically active. Quenching by iodide and measurement of fluorescence spectra, anisotropy, and lifetimes were used to characterize the response of the fluorescence of the probe in the peptide receptor complex for agonists and antagonists. All three fluorescence indicators provided the same insights into differences in the environment of the same indicator in the analogous position for agonist and antagonist peptides bound to the CCK receptor. Each agonist had its fluorescence quenched more easily and showed lower anisotropy (higher mobility of the probe) and shorter lifetime than the analogous antagonist. Treatment of agonist-occupied receptors with a non hydrolyzable GTP analogue shifted the receptor into its inactive low affinity state and increased probe fluorescence lifetimes toward values observed with antagonist probes. These data are consistent with a molecular conformational change associated with receptor activation that causes the amino terminus of the ligand (situated above transmembrane segment six) to move away from its somewhat protected environment and toward the aqueous milieu. PMID- 11893748 TI - A model for the stoichiometric regulation of blood coagulation. AB - We have developed a model of the extrinsic blood coagulation system that includes the stoichiometric anticoagulants. The model accounts for the formation, expression, and propagation of the vitamin K-dependent procoagulant complexes and extends our previous model by including: (a) the tissue factor pathway inhibitor (TFPI)-mediated inactivation of tissue factor (TF).VIIa and its product complexes; (b) the antithrombin-III (AT-III)-mediated inactivation of IIa, mIIa, factor VIIa, factor IXa, and factor Xa; (c) the initial activation of factor V and factor VIII by thrombin generated by factor Xa-membrane; (d) factor VIIIa dissociation/activity loss; (e) the binding competition and kinetic activation steps that exist between TF and factors VII and VIIa; and (f) the activation of factor VII by IIa, factor Xa, and factor IXa. These additions to our earlier model generate a model consisting of 34 differential equations with 42 rate constants that together describe the 27 independent equilibrium expressions, which describe the fates of 34 species. Simulations are initiated by "exposing" picomolar concentrations of TF to an electronic milieu consisting of factors II, IX, X, VII, VIIa, V, and VIIII, and the anticoagulants TFPI and AT-III at concentrations found in normal plasma or associated with coagulation pathology. The reaction followed in terms of thrombin generation, proceeds through phases that can be operationally defined as initiation, propagation, and termination. The generation of thrombin displays a nonlinear dependence upon TF, AT-III, and TFPI and the combination of these latter inhibitors displays kinetic thresholds. At subthreshold TF, thrombin production/expression is suppressed by the combination of TFPI and AT-III; for concentrations above the TF threshold, the bolus of thrombin produced is quantitatively equivalent. A comparison of the model with empirical laboratory data illustrates that most experimentally observable parameters are captured, and the pathology that results in enhanced or deficient thrombin generation is accurately described. PMID- 11893749 TI - Protein levels of Escherichia coli thioredoxins and glutaredoxins and their relation to null mutants, growth phase, and function. AB - Levels of Escherichia coli thioredoxin 1 (Trx1), Trx2, glutaredoxin 1 (Grx1), Grx2, and Grx3 have been determined by novel sensitive sandwich enzyme-linked immunosorbent assay. In a wild type strain, levels of Trx1 increased from the exponential to the stationary phase of growth (1.5-fold to 3400 ng/mg), as did levels of Grx2 (from approximately 2500 to approximately 8000 ng/mg). Grx3 and Trx2 levels were quite stable during growth ( approximately 4500 and approximately 200 ng/mg, respectively). Grx1 levels decreased from approximately 600 ng/mg at the exponential phase to approximately 285 ng/mg at the stationary phase. A large elevation of Grx1 (20-30-fold), was observed in null mutants for the thioredoxin system whereas levels of the other redoxins in all combinations of examined null mutants barely exceeded a 2-3-fold increase. Measurements of thymidine incorporation in newly synthesized DNA suggested that mainly Grx1 and, to a lesser extent, Trx1 contribute to the reduction of ribonucleotides. All glutaredoxin species were elevated in catalase-deficient strains, implying an antioxidant role for the glutaredoxins. Trx1, Trx2, and Grx1 levels increased after exposure to hydrogen peroxide and decreased after exposure to mercaptoethanol. The levels of Grx2 and Grx3 behaved exactly the opposite, suggesting that the transcription factor OxyR does not regulate their expression. PMID- 11893750 TI - Physical interaction with human tumor-derived p53 mutants inhibits p63 activities. AB - The p53 tumor suppressor gene is the most frequent target for genetic alterations in human cancers, whereas the recently discovered homologues p73 and p63 are rarely mutated. We and others have previously reported that human tumor-derived p53 mutants can engage in a physical association with different isoforms of p73, inhibiting their transcriptional activity. Here, we report that human tumor derived p53 mutants can associate in vitro and in vivo with p63 through their respective core domains. We show that the interaction with mutant p53 impairs in vitro and in vivo sequence-specific DNA binding of p63 and consequently affects its transcriptional activity. We also report that in cells carrying endogenous mutant p53, such as T47D cells, p63 is unable to recruit some of its target gene promoters. Unlike wild-type p53, the binding to specific p53 mutants markedly counteracts p63-induced growth inhibition. This effect is, at least partially, mediated by the core domain of mutant p53. Thus, inactivation of p53 family members may contribute to the biological properties of specific p53 mutants in promoting tumorigenesis and in conferring selective survival advantage to cancer cells. PMID- 11893751 TI - Yeast DNA damage-inducible Rnr3 has a very low catalytic activity strongly stimulated after the formation of a cross-talking Rnr1/Rnr3 complex. AB - The ribonucleotide reductase system in Saccharomyces cerevisiae includes four genes (RNR1 and RNR3 encoding the large subunit and RNR2 and RNR4 encoding the small subunit). RNR3 expression, nearly undetectable during normal growth, is strongly induced by DNA damage. Yet an rnr3 null mutant has no obvious phenotype even under DNA damaging conditions, and the contribution of RNR3 to ribonucleotide reduction is not clear. To investigate the role of RNR3 we expressed and characterized the Rnr3 protein. The in vitro activity of Rnr3 was less than 1% of the Rnr1 activity. However, a strong synergism between Rnr3 and Rnr1 was observed, most clearly demonstrated in experiments with the catalytically inactive Rnr1-C428A mutant, which increased the endogenous activity of Rnr3 by at least 10-fold. In vivo, the levels of Rnr3 after DNA damage never reached more than one-tenth of the Rnr1 levels. We propose that heterodimerization of Rnr3 with Rnr1 facilitates the recruitment of Rnr3 to the ribonucleotide reductase holoenzyme, which may be important when Rnr1 is limiting for dNTP production. In complex with inactive Rnr1-C428A, the activity of Rnr3 is controlled by effector binding to Rnr1-C428A. This result indicates cross-talk between the Rnr1 and Rnr3 polypeptides of the large subunit. PMID- 11893752 TI - Integrin activation involves a conformational change in the alpha 1 helix of the beta subunit A-domain. AB - The ligand-binding region of integrin beta subunits contains a von Willebrand factor type A-domain: an alpha/beta "Rossmann" fold containing a metal ion dependent adhesion site (MIDAS) on its top face. Although there is evidence to suggest that the betaA-domain undergoes changes in tertiary structure during receptor activation, the identity of the secondary structure elements that change position is unknown. The mAb 12G10 recognizes a unique cation-regulated epitope on the beta(1) A-domain, induction of which parallels the activation state of the integrin (i.e. competency for ligand recognition). The ability of Mn(2+) and Mg(2+) to stimulate 12G10 binding is abrogated by mutation of the MIDAS motif, demonstrating that the MIDAS is a Mn(2+)/Mg(2+) binding site and that occupancy of this site induces conformational changes in the A-domain. The cation-regulated region of the 12G10 epitope maps to Arg(154)/Arg(155) in the alpha1 helix. Our results demonstrate that the alpha1 helix undergoes conformational alterations during integrin activation and suggest that Mn(2+) acts as a potent activator of beta(1) integrins because it can promote a shift in the position of this helix. The mechanism of beta subunit A-domain activation appears to be distinct from that of the A-domains found in some integrin alpha subunits. PMID- 11893753 TI - Evaluation of the role of phosphatidylserine translocase activity in ABCA1 mediated lipid efflux. AB - The following two theories for the mechanism of ABCA1 in lipid efflux to apolipoprotein acceptors have been proposed: 1) that ABCA1 directly binds the apolipoprotein ligand and then facilitates lipid efflux and 2) that ABCA1 acts as a phosphatidylserine (PS) translocase, increasing PS levels in the plasma membrane exofacial leaflet, and that this is sufficient to facilitate apolipoprotein binding and lipid assembly. Upon induction of ABCA1 in RAW264.7 cells by cAMP analogues there was a moderate increase in cell surface PS as detected by annexin V binding, whereas apoAI binding was increased more robustly. Apoptosis induced large increases in annexin V and apoAI binding; however, apoptotic cells did not efflux lipids to apoAI. Annexin V did not act as a cholesterol acceptor, and it did not compete for the cholesterol acceptor or cell binding activity of apoAI. ApoAI binds to ABCA1-expressing cells, and with incubation at 37 degrees C apoAI is co-localized within the cells in ABCA1 containing endosomes. Fluorescent recovery after photobleaching demonstrated that apoAI bound to ABCA1-expressing cells was relatively immobile, suggesting that it was bound either directly or indirectly to an integral membrane protein. Although ABCA1 induction was associated with a small increase in cell surface PS, these results argue against the notion that this cell surface PS is sufficient to mediate cellular apoAI binding and lipid efflux. PMID- 11893755 TI - A ubiquitously expressed human hexacoordinate hemoglobin. AB - We have identified a new human hemoglobin that we call histoglobin because it is expressed in a wide array of tissues. Histoglobin shares less than 30% identity with the other human hemoglobins, and the gene contains an intron in an unprecedented location. Spectroscopic and kinetic experiments with recombinant human histoglobin indicate that it is a hexacoordinate hemoglobin with significantly different ligand binding characteristics than the other human hexacoordinate hemoglobin, neuroglobin. In contrast to the very high oxygen affinities displayed by most hexacoordinate hemoglobins, the biophysical characteristics of histoglobin indicate that it could facilitate oxygen transport. The discovery of histoglobin demonstrates that humans, like plants, differentially express multiple hexacoordinate hemoglobins. PMID- 11893754 TI - Normal assembly of 60 S ribosomal subunits is required for the signaling in response to a secretory defect in Saccharomyces cerevisiae. AB - A secretory defect leads to transcriptional repression of both ribosomal protein and rRNA genes in yeast. To elucidate the mechanism of the signaling, we previously isolated rrs mutants that were unable to respond to a secretory defect, and we cloned RRS1 encoding a nuclear protein that was required for ribosome biogenesis (Tsuno, A., Miyoshi, K., Tsujii, R., Miyakawa, T., and Mizuta, K. (2000) Mol. Cell. Biol. 20, 2066-2074). We identified duplicated genes encoding ribosomal protein L11, RPL11B as a wild-type allele complementing the rrs2 mutation, and RPL11A in two-hybrid screening using RRS1 as bait. Rpl11p was copurified with Rrs1p in immunoprecipitation analysis. Ultracentrifugation analysis revealed that Rrs1p associated fairly tightly with 60 S preribosomal subunits. These results suggest that signaling in response to a secretory defect requires the normal assembly of 60 S ribosomal subunits including Rrs1p and Rpl11p. PMID- 11893756 TI - Modification of late membrane permeability in avian reovirus-infected cells: viroporin activity of the S1-encoded nonstructural p10 protein. AB - Infection of chicken embryo fibroblasts by avian reovirus induces an increase in the permeability of the host plasma membrane at late, but not early, infection times. The absence of permeability changes at early infection times, as well as the dependence of late membrane modification on both viral protein synthesis and an active exocytic route, suggest that a virus-encoded membrane protein is required for avian reovirus to permeabilize cells. Further studies revealed that expression of nonstructural p10 protein in bacterial cells arrested cell growth and enhanced membrane permeability. Membrane leakiness was also observed following transient expression of p10 in BSC-40 monkey cells. Both its permeabilizing effect and the fact that p10 shares several structural and physical characteristics with other membrane-active viral proteins indicate that p10 is an avian reovirus viroporin. Furthermore, the fusogenic extracellular NH(2)-terminal domain of p10 appears to be dispensable for permeabilizing activity, because its deletion entirely abolished the fusogenic activity of p10, without affecting its ability to associate with cell membranes and to enhance membrane permeability. Similar properties have reported previously for immunodeficiency virus type I transmembrane glycoprotein gp41. Thus, like gp41, p10 appears to be a multifunctional protein that plays key roles in virus-host interaction. PMID- 11893757 TI - Why did some ichthyosaurs have such large eyes? AB - Many species of extinct marine ichthyosaurs had much larger eyes for their body size than would be expected of extant marine mammals and reptiles. Sensitivity to low light at great depth for the deep-diving genus Ophthalmosaurus has recently been suggested as the reason for the large eyes of these animals. Here, we discuss the implications for vision at such depths and consider other optical factors determining eye size. We suggest that the large eyes of ichthyosaurs are more likely to be the result of simultaneous selection for both sensitivity to low light and visual acuity. The importance of the evolutionary history of extant marine mammals and extinct ichthyosaurs is discussed, as are ecological factors driving both acuity and sensitivity. PMID- 11893758 TI - Hyperosmotic and thermal stresses activate p38-MAPK in the perfused amphibian heart. AB - We assessed the activation of p38-MAPK (mitogen-activated protein kinase) by osmotic and thermal stresses in the isolated perfused amphibian (Rana ridibunda) heart. Hyperosmotic stress induced the rapid activation of the kinase. In particular, in the presence of 0.5 mol l(-1) sorbitol, p38-MAPK was maximally phosphorylated (by approximately twelvefold) at 15 min, while excess of NaCl (206 mmol l(-1) final concentration) or KCl (16 mmol l(-1) final concentration) stimulated a less potent activation, maximised (by approximately eightfold and fourfold) within 2 min and 30 s, respectively, relative to control values. The effect of all three compounds examined was reversible, since the kinase phosphorylation levels decreased upon reperfusion of the heart with normal bicarbonate-buffered saline. Conversely, hypotonicity did not induce any p38-MAPK activation. Furthermore, both hypothermia and hyperthermia induced considerable phosphorylation of the kinase, by four- and 7.5-fold, respectively, relative to control values. Immunohistochemical studies elucidated the localisation pattern of phospho-p38-MAPK and also revealed enhanced atrial natriuretic peptide (ANP) immunoreactivity in osmotically stressed hearts. Interestingly, SB 203580 (1 micromol l(-1)) not only completely blocked the activation of p38-MAPK by all these interventions, but also abolished the enhanced ANP immunoreactivity induced by 0.5 mol l(-1) sorbitol. These findings indicate the possible involvement of ANP in the mechanisms regulating responses under such stressful conditions. PMID- 11893759 TI - Insect egg deposition induces Pinus sylvestris to attract egg parasitoids. AB - Plant volatiles released in response to feeding insects are known to attract enemies of the feeding herbivores. In this study, egg deposition by a herbivorous insect was shown to induce a gymnosperm plant to emit volatiles that attract egg parasitoids. Odour from twigs of Pinus sylvestris laden with egg masses of the pine sawfly Diprion pini attracts the eulophid egg parasitoid Chrysonotomyia ruforum. Volatiles released from pine twigs without diprionid eggs are not attractive. Oviposition by the sawfly onto pine needles induces not only a local response in pine needles laden with eggs but also a systemic reaction. Needles without eggs but adjacent to those bearing diprionid eggs also release the volatiles that attract the egg parasitoid. The elicitor of the attractive volatiles was shown to be present in the oviduct secretion coating the eggs of D. pini. When pine twigs are treated with jasmonic acid, a well-known plant wound signal, they emit volatiles that attract the egg parasitoid. These results show, for the first time, that a gymnosperm plant is able to attract parasitoids as soon as a herbivore has deposited its eggs on it. Thus, the plant appears to defend itself against herbivores prior to being damaged by feeding larvae. PMID- 11893761 TI - Heart rate and energy expenditure of incubating wandering albatrosses: basal levels, natural variation, and the effects of human disturbance. AB - We studied the changes in heart rate (HR) associated with metabolic rate of incubating and resting adult wandering albatrosses (Diomedea exulans) on the Crozet Islands. Metabolic rates of resting albatrosses fitted with external HR recorders were measured in a metabolic chamber to calibrate the relationship between HR and oxygen consumption (V(O(2))) (V(O(2))=0.074 x HR+0.019, r(2)=0.567, P<0.001, where V(O(2)) is in ml kg(-1) min(-1) and HR is in beats min(-1)). Incubating albatrosses were then fitted with HR recorders to estimate energy expenditure of albatrosses within natural field conditions. We also examined the natural variation in HR and the effects of human disturbance on nesting birds by monitoring the changes in HR. Basal HR was positively related to the mass of the individual. The HR of incubating birds corresponded to a metabolic rate that was 1.5-fold (males) and 1.8-fold (females) lower than basal metabolic rate (BMR) measured in this and a previous study. The difference was probably attributable to birds being stressed while they were held in the metabolic chamber or wearing a mask. Thus, previous measurements of metabolic rate under basal conditions or for incubating wandering albatrosses are likely to be overestimates. Combining the relationship between HR and metabolic rate for both sexes, we estimate that wandering albatrosses expend 147 kJ kg(-1) day(-1) to incubate their eggs. In addition, the cost of incubation was assumed to vary because (i) HR was higher during the day than at night, and (ii) there was an effect of wind chill (<0 degrees C) on basal HR. The presence of humans in the vicinity of the nest or after a band control was shown to increase HR for extended periods (2-3 h), suggesting that energy expenditure was increased as a result of the disturbance. Lastly, males and females reacted differently to handling in terms of HR response: males reacted more strongly than females before handling, whereas females took longer to recover after being handled. PMID- 11893760 TI - Insect midgut K(+) secretion: concerted run-down of apical/basolateral transporters with extra-/intracellular acidity. AB - In lepidopteran larvae, three transport mechanisms are involved in the active and electrogenic K(+) secretion that occurs in the epithelial goblet cells of the midgut. These consist of (i) basolateral K(+) channels, allowing K(+) entry from the haemolymph into the cytosol, (ii) apical electrogenic K(+)/2H(+) antiporters, which are responsible for secondary active extrusion of K(+) from the cell into the gut lumen via the goblet cavity and (iii) apical V-ATPase-type proton pumps. The latter energize apical K(+) exit by building up a large, cavity-positive electrical potential that drives the antiporters. Net K(+) secretion (I(K)) can be measured as short-circuit current (I(sc)) across the in vitro midgut mounted in an Ussing chamber. We investigated the influence of protons on the transepithelial I(K) and the partial reactions of the basolateral K(+) permeability (P(K)) and the apical, lumped 'K(+) pump' current (I(P)) at various extra- and intracellular pH values. In particular, we wanted to know whether increased cellular acidity could counteract the reversible dissociation of the V ATPase into its V(1) and V(o) parts, as occurs in yeast after glucose deprivation and in the midgut of Manduca sexta during starvation or moulting, thus possibly enhancing K(+) transport. When intact epithelia were perfused with high-[K(+)] (32 mmol l(-1)) salines with different pH values, I(K) was reversibly reduced when pH values fell below 6 on either side of the epithelium. Attempts to modify the intracellular pH by pulsing with NH(4)(+) or propionate showed that intracellular acidification caused a reduction in I(K) similar to that obtained in response to application of external protons. Treatment with azide, a well known inhibitor of the mitochondrial ATP synthase, had the same effect as pulsing with ammonium or propionate with, however, much faster kinetics and higher reversibility. Breakdown of the basolateral or apical barrier using the antibiotic nystatin allowed the intracellular pH to be clamped to that of the saline facing the nystatin-treated epithelial border. Cell acidification achieved by this manipulation led to a reduction in both apical I(P) and basolateral P(K). The transepithelial I(K) showed an approximately half-maximal reduction at external pH values close to 5 in intact tissues, and a similar reduction in I(P) and P(K) values was seen at an intracellular pH of 5 in nystatin-permeabilised epithelia. Thus, the hypothesized V(1)V(o) stabilization by cell acidity is not reflected in the pH-sensitivity of I(P). Moreover, all components that transport K(+) are synchronously inhibited below pH 6. The significance of our findings for the midgut in vivo is discussed. PMID- 11893762 TI - Visual control of host pursuit in the parasitoid fly Exorista japonica. AB - The tachinid fly Exorista japonica is a parasitoid of many kinds of lepidopterous larvae. After encountering a suitable host, the fly pursues the crawling larva on foot using visual cues to guide it. To investigate the visual control of host pursuit, we observed and videotaped pursuits of a host, the common armyworm Mythimna separata, for frame-by-frame analysis. Observation was performed in sunlight and under illumination from a fluorescent lamp. The fly pursued hosts discontinuously with a repeated stop-and-run motion. During a run, its movements consisted of rotation, forward translation and sideways translation. Rotation during a run was positively correlated with the angular position of the host's head. The direction of translation depended on the angular position of the host's head. Forward translation was negatively correlated with the visual angle subtended by the host. These results suggest that the fly orients and walks towards the leading edge of a moving target. There was little difference in the results between sunlight and illumination from a fluorescent lamp. PMID- 11893763 TI - Antagonistic control of fluid secretion by the Malpighian tubules of Tenebrio molitor: effects of diuretic and antidiuretic peptides and their second messengers. AB - Fluid secretion by insect Malpighian tubules is controlled by haemolymph-borne factors. The mealworm Tenebrio molitor provides the first known example of antagonistic interactions between endogenous neuropeptides acting on Malpighian tubules. The two corticotropin-releasing-factor (CRF)-related diuretic peptides previously isolated from Tenebrio molitor, Tenmo-DH(37) and Tenmo-DH(47), were found to stimulate Tenebrio molitor tubules in vitro in a dose-dependent manner with EC(50) values of 0.12 nmol l(-1) and 26 nmol l(-1) respectively. However, no synergistic or additive effect was observed when these two peptides were tested simultaneously. We then investigated antagonism between second messengers: dose response curves were constructed for stimulation of Tenebrio molitor tubules by cyclic AMP and their inhibition by cyclic GMP. When both cyclic nucleotides were included in the bathing Ringer, the stimulatory effect of cyclic AMP was neutralised by cyclic GMP. Similarly, the stimulatory effect of Tenmo-DH(37) was reversed on addition of an antidiuretic peptide (Tenmo-ADF), which was recently isolated from Tenebrio molitor and acts via cyclic GMP. The cardioacceleratory peptide CAP(2b), originally isolated from Manduca sexta, also increases intracellular cyclic GMP levels and inhibited fluid secretion by Tenebrio molitor tubules, with an EC(50) value of 85 nmol l(-1). This inhibitory effect was reversed by Tenmo-DH(37). Endogenous diuretic and antidiuretic peptides, effective at low concentrations and acting via antagonistic second messengers, have the potential for fine control of secretion rates in the Malpighian tubules of Tenebrio molitor. PMID- 11893765 TI - Myofibrillar protein isoform expression is correlated with synaptic efficacy in slow fibres of the claw and leg opener muscles of crayfish and lobster. AB - In the crayfish and lobster opener neuromuscular preparations of the walking legs and claws, there are regional differences in synaptic transmission even though the entire muscle is innervated by a single excitatory tonic motor neuron. The innervation of the proximal fibres produced larger excitatory postsynaptic potentials (EPSPs) than those of the central fibres. The amplitudes of the EPSPs in the distal fibres were intermediate between those of the proximal and central regions. These differences in EPSP amplitudes were correlated with differences in short-term facilitation between the three regions. When given a 10- or 20-pulse train of stimuli, the proximal fibres showed greater short-term facilitation initially, often followed by a maximization of short-term facilitation towards the end of a train. In contrast, the central fibres showed a linear increase in short-term facilitation throughout a stimulus train. The distal fibres showed intermediate short-term facilitation compared with the other two regions. Analysis of myofibrillar isoforms showed that levels of troponin-T(1) (TnT(1)), a 55 kDa isoform expressed in slow-tonic (S(2)) fibres, were correlated with synaptic properties. Proximal fibres had the highest levels of TnT(1), with lower levels in distal fibres; central fibres lacked TnT(1), which is characteristic of slow-twitch (S(1)) fibres. In addition, differences in troponin-I isoforms correlated with TnT(1) levels between the proximal, central and distal regions. The correlation between slow fibre phenotype and strength of innervation suggests a relationship between synaptic structure and expression of troponin isoforms. PMID- 11893764 TI - Physiological responses of Houbara bustards to high ambient temperatures. AB - Desert birds often experience a scarcity of drinking water and food and must survive episodes of high ambient temperature (T(a)). The physiological mechanisms that promote survival during extended periods of high T(a) have received little attention. We investigated the physiological responses of wild-caught and captive reared Houbara bustards, Chlamydotis macqueenii, to T(a) values ranging from below 0 degrees C to 55 degrees C, well above those in most previous studies of birds. Captive-reared Houbara bustards (mass 1245+/-242 g, N=7, mean +/- S.D.) in summer have a resting metabolic rate (RMR) of 261.4 kJ day(-1), 26 % below allometric predictions, and a total evaporative water loss (TEWL) at 25 degrees C of 25.8 g day(-1), 31 % below predictions. When T(a) exceeded body temperature (T(b)), the dry heat transfer coefficient decreased, a finding supporting the prediction that birds should minimize dry heat gain from the environment at high T(a) values. Houbara bustards withstand high T(a) values without becoming hyperthermic; at 45 degrees C, T(b) was on average 0.9 degrees C higher than at 25 degrees C. RMR and TEWL of captive-bred Houbara bustards were 23 % and 46 % higher in winter than in summer, respectively. Captive-reared Houbara bustards had a 17 % lower RMR and a 28 % lower TEWL than wild-born birds with similar genetic backgrounds. Differences in body composition between wild-caught and captive-reared birds were correlated with differences in physiological performance. PMID- 11893766 TI - Ion transport across posterior gills of hyperosmoregulating shore crabs (Carcinus maenas): amiloride blocks the cuticular Na(+) conductance and induces current noise. AB - Split gill lamellae and gill cuticles of shore crabs (Carcinus maenas) adapted to 10 per thousand salinity were mounted in a modified Ussing-type chamber. With NaCl saline on both sides, split gill lamellae generated a short-circuit current (I(sc)) of -301+/-16 microA cm(-2) at a conductance (G(te)) of 40+/-2 mS cm(-2). The net influxes of Na(+) and Cl(-) were 8.3+/-2.6 and 18.2+/-2.7 micromol cm(-2) h(-1), respectively. External amiloride (100 micromol l(-1)) reduced G(te) to approximately 50 % of the original value at unchanged I(sc); Cl(-) fluxes remained unaffected, whereas Na(+) fluxes were markedly reduced by 70-80 %. The I(sc) in the presence of external amiloride was almost completely inhibited by internal ouabain. At a clamp voltage of 50 mV (outside-positive), a positive current was measured at unchanged G(te). Under these conditions, amiloride reduced the current and conductance at half-maximal concentrations of 3.6 and 2.0 micromol l(-1), respectively. At outside-positive voltages, but not under short circuit conditions, external amiloride induced Lorentzian components in the power density spectra. The amiloride-dependent changes in the corner frequency (linear) and of the low-frequency plateau ('bell-shaped') were as expected for channel blockade by amiloride with pseudo-first-order kinetics. With an outside-positive clamp voltage of 50 mV across isolated cuticles, a positive cuticular current (I(cut)) of 25 188+/-3791 microA cm(-2) and a cuticular conductance (G(cut)) of 547+/-76 mS cm(-2) were measured. External amiloride reduced I(cut) and G(cut) at half-maximal concentrations of 0.7 and 0.6 micromol l(-1), respectively. Amiloride-induced current-noise analysis gave similar results to those observed with split gill lamellae. Ion-substitution experiments with isolated cuticles further support inhibition by external amiloride of the cuticular Na(+) conductance of shore crab gills and not amiloride-sensitive transporters (Na(+) channels or Na(+)/H(+) antiports) in the apical membrane. PMID- 11893767 TI - Mechanics of the respiratory system in the newborn tammar wallaby. AB - We investigated whether the mechanical properties of the respiratory system represent a major constraint to spontaneous breathing in the newborn tammar wallaby Macropus eugenii, which is born after a very short gestation (approximately 28 days, birth mass approximately 380 mg). The rate of oxygen consumption (V(O(2))) through the skin was approximately 33 % of the total V(O(2)) at day 1 and approximately 14 % at day 6. The mass-specific resting minute ventilation (E) and the ventilatory equivalent (VE/(O(2))) were approximately the same at the two ages, with a breathing pattern significantly deeper and slower at day 1. The mass-specific compliance of the respiratory system (C(rs)) did not differ significantly between the two age groups and was close to the values predicted from measurements in eutherian newborns. Mass specific respiratory system resistance (R(rs)) at day 1 was higher than at day 6, and also higher than in eutherian newborns. Chest distortion, quantified as the degree of abdominal motion during spontaneous breathing compared with that required to inflate the lungs passively, at day 1 was very large, whereas it was modest at day 6. We conclude that, in the tammar wallaby at birth, the high resistance of the respiratory system and the distortion of the chest wall greatly reduce the mechanical efficiency of breathing. At this age, gas exchange through the skin is therefore an important complement to pulmonary ventilation. PMID- 11893768 TI - Shell clamping behaviour in the limpet Cellana tramoserica. AB - The behaviour of clamping the shell against the substratum may play an important role in the limpet adhesion mechanism because friction generated by this behaviour resists dislodgement by shear forces. This paper describes the development of an apparatus to analyse limpet clamping activity in relation to known forces, including simulated wave activity and predator attack. The results show that Cellana tramoserica clamps its shell in a closely regulated manner consistent with an active role in the limpet adhesion mechanism. Limpets clamped sharply for several seconds in response to single disturbances such as tapping the shell. In response to more continuous disturbance simulating a concerted predator attack, limpets clamped tightly for several minutes. In response to lifting forces applied to the shell, limpets clamped at a set proportion of the lifting force, even if the lift force was a highly dynamic wave profile. This behaviour has implications for numerical models that attempt to describe limpet adhesion because it shows that limpets cannot be represented by a simple mechanical analogue and that the clamping behaviour must be accounted for if useful predictions are to be drawn. PMID- 11893769 TI - Template-matching describes visual pattern-recognition tasks in the weakly electric fish Gnathonemus petersii. AB - Several insects use template-matching systems to recognize objects or environmental landmarks by comparing actual and stored retinal images. Such systems are not viewpoint-invariant and are useful only when the locations in which the images have been stored and where they are later retrieved coincide. Here, we describe that a vertebrate, the weakly electric fish Gnathonemus petersii, appears to use template-matching to recognize visual patterns that it had previously viewed from a fixed vantage point. This fish is nocturnal and uses its electrical sense to find its way in the dark, yet it has functional vision that appears to be well adapted to dim light conditions. We were able to train three fish in a two-alternative forced-choice procedure to discriminate a rewarded from an unrewarded visual pattern. From its daytime shelter, each fish viewed two visual patterns placed at a set distance behind a transparent Plexiglas screen that closed the shelter. When the screen was lifted, the fish swam towards one of the patterns to receive a food reward or to be directed back into its shelter. Successful pattern discrimination was limited to low ambient light intensities of approximately 10 lx and to pattern sizes subtending a visual angle greater than 3 degrees. To analyze the characteristics used by the fish to discriminate the visual training patterns, we performed transfer tests in which the training patterns were replaced by other patterns. The results of all such transfer tests can best be explained by a template-matching mechanism in which the fish stores the view of the rewarded training pattern and chooses from two other patterns the one whose retinal appearance best matches the stored view. PMID- 11893770 TI - Discrimination of closed shapes by two species of bee, Apis mellifera and Megachile rotundata. AB - In the present study, the performance of two bee species, the honeybee Apis mellifera and the leaf-cutter bee Megachile rotundata, in discriminating among various closed (convex) shapes was examined systematically for the first time. Bees were trained to each of five different shapes, a disc, a square, a diamond and two different triangles, all of the same area, using fresh bees in each experiment. In subsequent tests, the trained bees were given a choice between the learned shape and each of the other four shapes. Two sets of experiments were conducted with both species. In the first, solid black shapes were presented against a white background, thus providing a high luminance contrast. In the second, the shapes carried a random black-and-white pattern and were presented 5 cm in front of a similar pattern, thus producing motion contrast, rather than luminance contrast, against the background. The results obtained with the solid shapes reveal that both bee species accomplish the discrimination, although the performance of the honeybee is significantly better than that of the leaf-cutter bee. Furthermore, the effectiveness of the various shapes differs between the two species. However, in neither species is the discrimination performance correlated with the amount of overlap of the black areas contained in the various pairs of shapes, suggesting that, in our experiments, shape discrimination is not based on a template-matching process. We propose that it is based on the use of local parameters situated at the outline of the shape, such as the position of angles or acute points and, in particular, the position and orientation of edges. This conclusion is supported by the finding that bees of both species accomplish the discrimination even with the patterned shapes. These shapes are visible only because of the discontinuity of the speed of image motion perceived at the edge between the shape and the background. PMID- 11893771 TI - Regulation of xenobiotic and bile acid metabolism by the nuclear pregnane X receptor. AB - The nuclear pregnane X receptor (PXR; NR1I2) is an integral component of the body's defense mechanism against chemical insult (chemoprotection). PXR is activated by a diverse array of lipophilic chemicals, including xenobiotics and endogenous substances, and regulates the expression of cytochromes P450, conjugating enzymes, and transporters involved in the metabolism and elimination of these potentially harmful chemicals from the body. Among the chemicals that bind and activate PXR is the toxic bile acid lithocholic acid; activation of PXR, in turn, protects against the severe liver damage caused by this bile acid.Thus, PXR serves as a physiological sensor of lithocholic acid and perhaps other bile acids and coordinately regulates genes involved in their detoxification. Interestingly, both the antibiotic rifampicin and the herbal antidepressant St. John's wort activate PXR and have anticholestatic properties, which suggests that more potent, selective PXR agonists may be useful in the treatment of biliary cholestasis or other diseases characterized by the accumulation of bile acids or other toxins in the liver. PMID- 11893772 TI - Properties of triglyceride-rich and cholesterol-rich lipoproteins in the remnant like particle fraction of human blood plasma. AB - An immunoassay procedure that quantifies remnant-like particle (RLP) cholesterol in human blood plasma has shown considerable promise as a clinically applicable risk marker for atherosclerotic disease. The lipoproteins included in this assay include not only certain TG-rich lipoproteins [all particles containing apolipoprotein B-48 (apoB-48) and a fraction of those containing apoB-100] but also a very small proportion of plasma cholesterol-rich lipoproteins. The TG-rich lipoprotein component of RLP has been partially characterized, but relatively little is known about the component cholesterol-rich lipoproteins. We have further characterized the properties of the TG-rich component that is included in RLP in which about 25% of the particles contain apoB-48 and the remainder apoB 100. We show that the cholesterol-rich component is comprised mainly of beta migrating LDLs that contain predominantly apoB-100. ApoE found in the LDL fraction of RLP resides on pre-beta lipoproteins that lack apoA-I as well as apoB. The TG-rich component of RLP is responsible for increased RLP-cholesterol concentrations associated with hypertriglyceridemia. By contrast, the cholesterol rich component is a major contributor to plasma RLP-cholesterol in individuals with low plasma TG. Our results suggest that particle heterogeneity in the RLP fraction is likely to affect the ability of RLP-cholesterol concentration to predict atherosclerotic risk. RLP-cholesterol concentrations in individuals with low plasma TG may not have the same clinical significance as they do in those with hypertriglyceridemia. PMID- 11893773 TI - ApoE-dependent sterol efflux from macrophages is modulated by scavenger receptor class B type I expression. AB - Macrophages express a number of proteins involved in sterol efflux pathways, including apolipoprotein E (apoE) and scavenger receptor class B type I (SR-BI). We have investigated a potential interaction between these two sterol efflux pathways in modulating overall macrophage sterol flux. We utilized an experimental system in which we increased expression of each of these proteins to a high physiologic range in order to perform our evaluation. We show that in apoE expressing cells, a 4-fold increase in SR-BI expression leads to reduction of sterol and phospholipid efflux. SR-BI-mediated reduction in sterol efflux was only observed in cells that expressed endogenous apoE. In J774 cells that did not express apoE, a similar increase in SR-BI level led to increased sterol efflux. The divergent response of sterol efflux after increased SR-BI was maintained in the presence of a number of structurally diverse extracellular sterol acceptors. Increased SR-BI expression also enhanced sterol efflux to exogenously added apoE. Investigation of a potential mechanism for reduced efflux in apoE-expressing cells indicated that SR-BI expression reduced macrophage apoE by accelerating the degradation of newly synthesized apoE. This led to decreased secretion of apoE and reduced the fraction of apoE sequestered on the cell surface. Thus, enhanced SR-BI expression in macrophages can reduce the cellular level and secretion of apoE by accelerating degradation of the newly synthesized protein. This reduction of endogenous apoE is accompanied by reduced sterol efflux from macrophages. PMID- 11893774 TI - Estrogen receptor-mediated repression of human hepatic lipase gene transcription. AB - Estrogen replacement therapy in women decreases hepatic lipase (HL) activity, which may account for the associated increase in HDL cholesterol. To investigate whether estrogen decreases HL transcription, transient cotransfection assays with HL promoter and estrogen receptor-alpha (ERalpha) expression constructs were performed in HepG2 cells. 17beta-estradiol (E(2)) decreased transcription driven by the -1557/+41 human HL promoter by up to 50% at 10(-7) M. Mutation of ERalpha by deletion of its transactivation domains or ligand-binding domain eliminated E(2)-induced repression of the promoter, whereas deletion of the DNA-binding domain of ERalpha resulted in a 7-fold activation by E(2). The E(2)-induced repression was maintained after mutation of a potential estrogen-response element in the promoter. The region of estrogen responsiveness was localized to -1557/ 1175 of the HL promoter by deletion analysis. Mutation of an AP-1 site at -1493 resulted in a partial loss of E(2)-induced repression, similar to that caused by deletion of nucleotides -1557 to -1366. Gel shift assays with nuclear extracts from E(2)-treated HepG2 cells stably expressing ERalpha demonstrated an increase in binding to an AP-1 consensus oligonucleotide. The AP-1 activator, phorbol 12 myristate 13-acetate, inhibited the HL promoter by greater than 50%. Collectively, the data suggest that estrogen represses the transcription of the HL gene, possibly through an AP-1 pathway. PMID- 11893775 TI - LCAT facilitates transacylation of 17 beta-estradiol in the presence of HDL3 subfraction. AB - It has been shown that estrogens need to be metabolized to their hydrophobic estrogen ester derivatives to act as antioxidants in lipoproteins. Data suggest that 17beta-estradiol (E(2)) becomes esterified in LCAT-induced reactions and the esters are transported from HDL particles to LDL and VLDL particles by a CETP dependent mechanism. In the present study we have further investigated the regulation of E(2) esterification by LCAT and focused on the importance of HDL structure and composition in the esterification process. Isolated LDL, HDL(2), HDL(3), and reconstituted discoidal HDL (rHDL) were incubated with labeled E(2), with and without purified LCAT, at 37 degrees C for 24 h. After purification of the lipoprotein fractions, there was a significant peak of radioactivity representing esterified estradiol attached to HDL(3) and rHDL, but HDL(2) and LDL contained only trace amounts of labeled estradiol ester. TLC analysis confirmed that the radioactivity migrated in a position corresponding to that of 17beta E(2) 17-monoester standard. The amount of radioactivity associated with HDL(3) and rHDL representing esterified E(2) was significantly increased by addition of purified LCAT. However, only limited increases of radioactivity were observed in HDL(2) and LDL. In conclusion, HDL subfractions differ in their potential to regulate estradiol esterification by LCAT. PMID- 11893776 TI - Structural and functional consequences of missense mutations in exon 5 of the lipoprotein lipase gene. AB - Missense mutations in exon 5 of the LPL gene are the most common reported cause of LPL deficiency. Exon 5 is also the region with the strongest homology to pancreatic and hepatic lipase, and is conserved in LPL from different species. Mutant LPL proteins from post-heparin plasma from patients homozygous for missense mutations at amino acid positions 176, 188, 194, 205, and 207, and from COS cells transiently transfected with the corresponding cDNAs were quantified and characterized, in an attempt to determine which aspect of enzyme function was affected by each specific mutation. All but one of the mutant proteins were present, mainly as partially denatured LPL monomer, rendering further detailed assessment of their catalytic activity, affinity to heparin, and binding to lipoprotein particles difficult. However, the fresh unstable Gly(188)-->Glu LPL and the stable Ile(194)-->Thr LPL, although in native conformation, did not express lipase activity. It is proposed that many of the exon 5 mutant proteins are unable to achieve or maintain native dimer conformation, and that the Ile(194)-->Thr substitution interferes with access of lipid substrate to the catalytic pocket. These results stress the importance of conformational evaluation of mutant LPL. Absence of catalytic activity does not necessarily imply that the substituted amino acid plays a specific direct role in catalysis. PMID- 11893777 TI - Replication of linkage of familial hypobetalipoproteinemia to chromosome 3p in six kindreds. AB - Familial hypobetalipoproteinemia (FHBL) is a genetically heterogeneous condition characterized by very low apolipoprotein B (apoB) concentrations in plasma and/or low levels of LDL-cholesterol (LDL-C) with a propensity to developing fatty liver. In a minority of cases, truncation-specifying mutations of the apoB gene (APOB) are etiologic, but the genetic basis of most cases is unknown. We previously reported linkage of FHBL to a 10 cM region on 3p21.1-22 in one kindred. The objectives of the current study were to identify other FHBL families with linkage to 3p and to narrow the FHBL susceptibility region on 3p. Six additional FHBL kindreds unlinked to the APOB region on chromosome 2 were genotyped with polymorphic markers spanning a region of approximately 13 cM on chromosome 3. Quantitative linkage analyses indicated that the FHBL in these families was linked to 3p21.1-22. Haplotype analysis identified several meiotic crossover events, allowing us to narrow the critical region from 10 cM to 2.0 cM, between markers D3S2407 and D3S1767. PMID- 11893778 TI - Associations of LPL and APOC3 gene polymorphisms on plasma lipids in a Mediterranean population: interaction with tobacco smoking and the APOE locus. AB - We conducted a cross-sectional study in a Spanish population (n = 1,029) to investigate associations between the LPL and APOC3 gene loci (LPL-HindIII, LPL S447X, and APOC3-SstI) and plasma lipid levels and their interaction with APOE polymorphisms and smoking. Carriers of the H(-) or the X447 allele had higher levels of HDL cholesterol (HDL-C), and lower levels of TG, after adjustment for age, body mass index, alcohol, smoking, exercise, and education (P < 0.01). The APOC3 polymorphism presented additive effects to the LPL variants on TG and HDL-C levels in men, and on TG in women. The most and the least favorable haplotype combinations were H(-)/X447/S1 and H(+)/S447/S2, respectively. These combinations accounted for 7% and 5% of the variation in HDL-C and TG in men, and 3% and 4% in women. There was a significant interaction between APOE and LPL variants and HDL C levels in both genders (P < 0.05). The increases in HDL-C observed for the rare alleles were higher in epsilon4 than in epsilon3 subjects, and absent in epsilon2 individuals. This effect was modulated by smoking (interaction HindIII-APOE smoking, P = 0.019), indicating that smoking abolished the increase in HDL-C levels observed in epsilon4/H(-) subjects. Understanding this gene-gene environmental interaction may facilitate preventive interventions to reduce coronary artery disease risk. PMID- 11893779 TI - In vivo contribution of LCAT to apolipoprotein B lipoprotein cholesteryl esters in LDL receptor and apolipoprotein E knockout mice. AB - Previous studies have indicated that LCAT may play a role in the generation of cholesteryl esters (CE) in plasma apolipoprotein B (apoB) lipoproteins. The purpose of the present study was to examine the quantitative importance of LCAT on apoB lipoprotein CE fatty acid (CEFA) composition. LCAT(-/-) mice were crossed into the LDL receptor (LDLr)(-/-) and apoE(-/-) background to retard the clearance and increase the concentration of apoB lipoprotein in plasma. Plasma free cholesterol was significantly elevated but total and esterified cholesterol concentrations were not significantly affected by removal of functioning LCAT in either the LDLr(-/-) or apoE(-/-) mice consuming a chow diet. However, when functional LCAT was removed from LDLr(-/-) mice, the CEFA ratio (saturated + monounsaturated/polyunsaturated) of plasma LDL increased 7-fold because of a 2 fold increase in saturated and monounsaturated CE, a 40% reduction in cholesteryl linoleate, and a complete absence of long chain (>18 carbon) polyunsaturated CE (20:4, 20:5n-3, and 22:6n-3), from 29.3% to 0%. Removal of functional LCAT from apoE(-/-) mice resulted in only a 1.6-fold increase in the CEFA ratio, due primarily to a complete elimination of long chain CE (7.7% to 0%). Our results demonstrate that LCAT contributes significantly to the CEFA pool of apoB lipoprotein and is the only source of plasma long chain polyunsaturated CE in these mice. PMID- 11893780 TI - Stereochemistry of the peroxisomal branched-chain fatty acid alpha- and beta oxidation systems in patients suffering from different peroxisomal disorders. AB - Phytanic acid (3,7,11,15-tetramethylhexadecanoic acid) is a branched-chain fatty acid derived from dietary sources and broken down in the peroxisome to pristanic acid (2,6,10,14-tetramethylpentadecanoic acid) via alpha-oxidation. Pristanic acid then undergoes beta-oxidation in peroxisomes. Phytanic acid naturally occurs as a mixture of (3S,7R,11R)- and (3R,7R,11R)-diastereomers. In contrast to the alpha-oxidation system, peroxisomal beta-oxidation is stereospecific and only accepts (2S)-isomers. Therefore, a racemase called alpha-methylacyl-CoA racemase is required to convert (2R)-pristanic acid into its (2S)-isomer. To further investigate the stereochemistry of the peroxisomal oxidation systems and their substrates, we have developed a method using gas-liquid chromatography-mass spectrometry to analyze the isomers of phytanic, pristanic, and trimethylundecanoic acid in plasma from patients with various peroxisomal fatty acid oxidation defects. In this study, we show that in plasma of patients with a peroxisomal beta-oxidation deficiency, the relative amounts of the two diastereomers of pristanic acid are almost equal, whereas in patients with a defect of alpha-methylacyl-CoA racemase, (2R)-pristanic acid is the predominant isomer. Furthermore, we show that in alpha-methylacyl-CoA racemase deficiency, not only pristanic acid accumulates, but also one of the metabolites of pristanic acid, 2610-trimethylundecanoic acid, providing direct in vivo evidence for the requirement of this racemase for the complete degradation of pristanic acid. PMID- 11893781 TI - Effect of hydrogenated and saturated, relative to polyunsaturated, fat on immune and inflammatory responses of adults with moderate hypercholesterolemia. AB - Consumption of diets high in hydrogenated fat/trans fatty acids has been shown to have an adverse affect on lipoprotein profiles with respect to cardiovascular disease risk. Dietary fat and cholesterol play an important role in the regulation of immune and inflammatory responses shown to be involved in atherogenesis. We investigated the effects of diets containing hydrogenated fat on cellular immune response and production of inflammatory cytokines in human subjects with moderately elevated cholesterol levels (LDL cholesterol >130 mg/dl). In a double blind cross-over study, 19 subjects consumed three diets, 30% of calories as fat, of which two thirds were provided as soybean oil, soybean oil based stick margarine, or butter for 32 days, each in a randomized order. Production of proinflammatory mediators, prostaglandin (PG)E(2), interleukin (IL) 1beta, IL-6, and tumor necrosis factor alpha (TNF-alpha); delayed type hypersensitivity (DTH) response, in vitro lymphocyte proliferation, and production of IL-2 were determined. Production of IL-6 and TNF-alpha was significantly higher after consumption of stick margarine diet compared with soybean oil diet. IL-1beta and TNF-alpha production correlated positively with ratios of total cholesterol to HDL cholesterol (r = 0.499, P < 0.001 and r = 0.291, P = 0.04, respectively). There was no significant difference in DTH response, lymphocyte proliferation, or levels of IL-2 and PGE(2) produced among three groups. Our results indicate that consumption of a diet high in hydrogenated fat does not adversely affect cellular immunity but increases production of inflammatory cytokines that have been associated with the pathophysiology of atherosclerosis. PMID- 11893782 TI - LDL particle subclasses in hypercholesterolemia. Molecular determinants of reduced lipid hydroperoxide stability. AB - Hypercholesterolemia is characterized by elevated plasma levels of LDL in which the cholesteryl ester (CE)-rich LDL subclasses of light and intermediate density (LDL1+2 and LDL3, respectively) typically predominate. The molecular mechanisms implicated in oxidation of LDL particle subclasses in hypercholesterolemia are indeterminate. Lipid hydroperoxides (LOOH), primary oxidation products in LDL, are implicated in atherogenesis. LOOH formation was evaluated in light (LDL1+2), intermediate (LDL3), and dense (LDL4+5) LDL subclasses from hypercholesterolemic (HC) subjects (n = 7) during copper-mediated oxidative stress, and compared with that in corresponding subclasses from normolipidemic subjects (n = 7). HC LDL subclasses were distinguished by lower polyunsaturated phospholipid-alpha tocopherol ratios (P < 0.02), lower contents of phosphatidyl choline (PC)16:0 18:0/18:2 and PC16:0-18:0/20:4+22:6 (P < 0.002), and higher surface phospholipid free cholesterol ratios (P < 0.04). The LDL3, LDL4, and LDL5 subclasses in HC subjects displayed low-core polyunsaturated CE-alpha-tocopherol ratios (P < 0.05), despite similar PUFA CE content. These physicochemical differences did not modify the oxidative susceptibility of HC LDL but underlie the marked instability of cholesterol linoleate hydroperoxides in HC LDL1+2, LDL3, and LDL4 subclasses. Elevated concentrations of large, CE-rich, light, and intermediate LDL subclasses (LDL1+2, LDL3) in hypercholesterolemia may therefore act as an abundant proatherogenic source of highly unstable LOOH in the arterial wall. PMID- 11893783 TI - Role of ether-linked lysophosphatidic acids in ovarian cancer cells. AB - Naturally occurring alkyl- and alkenyl-lysophosphatidic acids (al-LPAs) are detected and elevated in ovarian cancer ascites compared with ascites from non malignant diseases. Here we describe the biological functions and signaling properties of these ether-linked LPAs in ovarian cancer cells. They are elevated and stable in ovarian cancer ascites, which represents an in vivo environment for ovarian cancer cells. They stimulated DNA synthesis and proliferation of ovarian cancer cells. In addition, they induced cell migration and the secretion of a pro angiogenic factor, interleukin-8 (IL-8), in ovarian cancer cells. The latter two processes are potentially related to tumor metastasis and angiogenesis, respectively. Al-LPAs induced diverse signaling pathways in ovarian cancer cells. Their mitogenic activity depended on the activation of the G(i/o) protein, phosphatidylinositol-3 kinase (PI3K), and mitogen-activated protein (MAP) kinase kinase (MEK), but not p38 mitogen activated protein kinase (MAP kinase). S473 phosphorylation of protein kinase B (Akt) by these lipids required activation of the G(i/o) protein, PI3K, MEK, p38 MAP kinase, and Rho. However, T308 phosphorylation of Akt stimulated by al-LPAs did not require activation of p38 MAP kinase. On the other hand, cell migration induced by al-LPAs depended on activities of the G(i/o) protein, PI3K, and Rho, but not MEK. These data suggest that ether-linked LPAs may play an important role in ovarian cancer development. PMID- 11893784 TI - Altered activities of anti-atherogenic enzymes LCAT, paraoxonase, and platelet activating factor acetylhydrolase in atherosclerosis-susceptible mice. AB - We examined whether the putative anti-atherogenic enzymes LCAT, paraoxonase (PON), and platelet-activating factor acetylhydrolase (PAF-AH) are impaired in 8 week old atherosclerosis susceptible apolipoprotein E (apoE)(-/-) and LDL receptor (LDLr)(-/-) mice and whether plasma concentrations of bioactive oxidized phospholipids accumulate in plasma. ApoE(-/-) mice had reduced (28%) LCAT activity and elevated lysophosphatidylcholine and bioactive oxidized phospholipids (1-palmitoyl-2-oxovaleryl-sn-glycero-3-phosphocholine and 1 palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine) compared with controls on the chow diet. Elevated oxidized phospholipids and reduced LCAT activity may, in part, contribute to spontaneous lesions in these mice on a chow diet. A Western diet decreased LCAT activity further (50% of controls) and PON activity was decreased 38%. The LDLr(-/-) mice showed normal LCAT activity on chow diet and little accumulation of oxidized phospholipids. On a Western diet, LDLr(-/-) mice had reduced LCAT activity (21%), but no change in PON activity. All genotypes had reduced PAF-AH activity on the Western diet. ApoE(-/-) and LDLr(-/-) mice, but not controls, had elevated plasma bioactive oxidized phospholipids on the Western diet. We conclude that impairment of LCAT activity and accumulation of oxidized phospholipids are part of an early atherogenic phenotype in these models. PMID- 11893785 TI - Heritability of plasma noncholesterol sterols and relationship to DNA sequence polymorphism in ABCG5 and ABCG8. AB - The plasma concentrations of cholesterol precursor sterols and plant sterols vary over a 5- to 10-fold range among normolipidemic individuals, and provide indices of the relative rates of cholesterol synthesis and fractional absorption. In the present study, we examined the relative contributions of genetic and environmental factors to variation in the plasma concentrations and sterol cholesterol ratios of five noncholesterol sterols, including the 5alpha-saturated derivative of cholesterol (cholestanol), two precursors in the cholesterol biosynthesis pathway (desmosterol and lathosterol), and two phytosterols (campesterol and sitosterol). Plasma sterol concentrations were highly stable in 30 individuals measured over a 48 week period. Regression of offspring sterol levels on the parental values indicated that plasma levels of all five noncholesterol sterols were highly heritable. Analysis of monozygotic and dizygotic twin pairs also indicated strong heritability of all five sterols. Two common sequence variations (D19H and T400K) in ABCG8, an ABC half-transporter defective in sitosterolemia, were associated with lower concentrations of plant sterols in parents, and in their offspring.Taken together, these findings indicate that variation in the plasma concentrations of noncholesterol sterols is highly heritable, and that polymorphism in ABCG8 contributes to genetic variation in the plasma concentrations of plant sterols. PMID- 11893786 TI - Identification of metabolites from type III F2-isoprostane diastereoisomers by mass spectrometry. AB - F(2)-isoprostanes (F(2)-iPs) are prostaglandin (PG)-like products of non enzymatic free radical-catalyzed peroxidation of arachidonic acid that are now widely used as indices of lipid peroxidation in vivo. Knowledge of the metabolic fate of F(2)-iPs in vivo is still scant, despite its importance for defining their overall formation and biological effects in vivo. Type III F(2)-iPs, which are diastereoisomers of cyclooxygenase-derived PGF(2alpha), may be metabolized through the pathways of PG metabolism. We therefore studied the in vitro metabolism of eight synthetic Type III F(2)-iP diastereoisomers in comparison with PGF(2alpha). We used gas chromatography-mass spectrometry and high performance liquid chromatography-electrospray-tandem mass spectrometry for structural identification of metabolites formed after incubation of the various compounds with isolated rat hepatocytes. PGF(2alpha) was metabolized to several known products, resulting from a combination of beta-oxidation, reduction of Delta(5) and/or Delta(13) double bonds, and 15-OH oxidation, plus other novel products deriving from conjugation with taurine of PGF(2alpha) and its metabolites. Of the eight F(2)-iP diastereoisomers, some were processed similarly to PGF(2alpha), whereas others showed peculiar metabolic profiles according to specific stereochemical configurations. These data represent the first evidence of biodegradation of selected Type III F(2)-iP isomers other than 8-epi PGF(2alpha), through known and novel pathways of PGF(2alpha) metabolism. The analytical characterization of these products may serve as a basis for identifying the most significant products formed in vivo. PMID- 11893787 TI - Coupled assay of sphingomyelin and ceramide molecular species by gas liquid chromatography. AB - This study reports a single-step analysis of the molecular species of endogenous ceramides and of the ceramide moiety of sphingomyelins in biological samples, using gas liquid chromatography (GLC). Silylated sphingomyelins were quantitatively converted to monosilylated ceramide upon injection into GLC, whereas the free ceramides were di-silylated on the primary and secondary alcohol function, as confirmed by mass spectrometry. The reproducible shift of the retention times between the mono- and di-silylated derivatives enables simultaneous quantification of the variety of sphingomyelin and ceramide molecular species. Overlapping diacylglycerols were first removed by a mild alkaline treatment of the lipid extract. The lowest detection limit (5 pmol) did not allow for identification of free ceramides in human plasma, but 17 molecular species of ceramides derived from sphingomyelins were quantified, from NC16:0 up to NC24:1. By contrast, three major free ceramides (NC16:0, NC24:0, and NC24:1) were quantified in HEPG2 and Chinese hamster ovary (CHO) cells. Upon induction of apoptosis in CHO cells by C6-ceramide, we could follow the disappearance of the C6-ceramide, its partial conversion to C6-sphingomyelin, and the prominent increase of NC16:0 ceramide. Thus, our method represents a unique procedure of simultaneous analysis of sphingomyelin and ceramide molecular species able to monitor the variation of the different pools in biological samples. PMID- 11893788 TI - UV analysis of Amadori-glycated phosphatidylethanolamine in foods and biological samples. AB - Maillard reactions are among the most important of the chemical and oxidative changes occurring in food and biological samples that contribute to food deterioration and to the pathophysiology of human disease. Although the association of lipid glycation with this process has recently been shown, the number of lipid glycation products in food and biological materials has not been clear. In this study, we synthesized the Amadori products derived from the glycation of phosphatidylethanolamine (PE), i.e., Amadori-PEs. Dioleoyl PE was incubated with glucose and lactose for 15 days, and the resultant Amadori-PEs were purified and isolated using solid phase extraction followed by HPLC. With this procedure, essentially pure (>98% purity) Amadori-PEs glycated with glucose (Glc-PE) and with lactose (Lac-PE) were obtained and used as standards in the subsequent studies. To determine the presence of Amadori-PEs in food and biological samples, the carbonyl group of Amadori-PEs was ultraviolet (UV) labeled with 3-methyl-2-benzothiazolinone hydrazone, and the labeled Amadori-PEs were analyzed with normal phase HPLC-UV (318 nm). The detection limit was 4.5 ng (5 pmol) for Glc-PE and 5.3 ng (5 pmol) for Lac-PE. Among the several food samples examined, infant formula and chocolate contained a high amount of both Glc-PE and Lac-PE over wide concentration ranges, such as 1.5-112 microg/g. Testing biological materials showed Amadori-PE (Glc-PE) was detectable in rat plasma. PMID- 11893789 TI - The prescription-drug problem. PMID- 11893790 TI - Exercise capacity and mortality among men referred for exercise testing. AB - BACKGROUND: Exercise capacity is known to be an important prognostic factor in patients with cardiovascular disease, but it is uncertain whether it predicts mortality equally well among healthy persons. There is also uncertainty regarding the predictive power of exercise capacity relative to other clinical and exercise test variables. METHODS: We studied a total of 6213 consecutive men referred for treadmill exercise testing for clinical reasons during a mean (+/-SD) of 6.2+/ 3.7 years of follow-up. Subjects were classified into two groups: 3679 had an abnormal exercise-test result or a history of cardiovascular disease, or both, and 2534 had a normal exercise-test result and no history of cardiovascular disease. Overall mortality was the end point. RESULTS: There were a total of 1256 deaths during the follow-up period, resulting in an average annual mortality of 2.6 percent. Men who died were older than those who survived and had a lower maximal heart rate, lower maximal systolic and diastolic blood pressure, and lower exercise capacity. After adjustment for age, the peak exercise capacity measured in metabolic equivalents (MET) was the strongest predictor of the risk of death among both normal subjects and those with cardiovascular disease. Absolute peak exercise capacity was a stronger predictor of the risk of death than the percentage of the age-predicted value achieved, and there was no interaction between the use or nonuse of beta-blockade and the predictive power of exercise capacity. Each 1-MET increase in exercise capacity conferred a 12 percent improvement in survival. CONCLUSIONS: Exercise capacity is a more powerful predictor of mortality among men than other established risk factors for cardiovascular disease. PMID- 11893791 TI - Prevalence of impaired glucose tolerance among children and adolescents with marked obesity. AB - BACKGROUND: Childhood obesity, epidemic in the United States, has been accompanied by an increase in the prevalence of type 2 diabetes among children and adolescents. We determined the prevalence of impaired glucose tolerance in a multiethnic cohort of 167 obese children and adolescents. METHODS: All subjects underwent a two-hour oral glucose-tolerance test (1.75 g [DOSAGE ERROR CORRECTED] of glucose per kilogram of body weight), and glucose, insulin, and C-peptide levels were measured. Fasting levels of proinsulin were obtained, and the ratio of proinsulin to insulin was calculated. Insulin resistance was estimated by homeostatic model assessment, and beta-cell function was estimated by calculating the ratio between the changes in the insulin level and the glucose level during the first 30 minutes after the ingestion of glucose. RESULTS: Impaired glucose tolerance was detected in 25 percent of the 55 obese children (4 to 10 years of age) and 21 percent of the 112 obese adolescents (11 to 18 years of age); silent type 2 diabetes was identified in 4 percent of the obese adolescents. Insulin and C-peptide levels were markedly elevated after the glucose-tolerance test in subjects with impaired glucose tolerance but not in adolescents with diabetes, who had a reduced ratio of the 30-minute change in the insulin level to the 30 minute change in the glucose level. After the body-mass index had been controlled for, insulin resistance was greater in the affected cohort and was the best predictor of impaired glucose tolerance. CONCLUSIONS: Impaired glucose tolerance is highly prevalent among children and adolescents with severe obesity, irrespective of ethnic group. Impaired oral glucose tolerance was associated with insulin resistance while beta-cell function was still relatively preserved. Overt type 2 diabetes was linked to beta-cell failure. PMID- 11893792 TI - Changes in mitochondrial DNA as a marker of nucleoside toxicity in HIV-infected patients. AB - BACKGROUND: Nucleoside analogues can induce toxic effects on mitochondria by inhibiting the human DNA polymerase gamma. The toxic effects can range from increased serum lactate levels to potentially fatal lactic acidosis. We studied changes in mitochondrial DNA relative to nuclear DNA in the peripheral-blood cells of patients with symptomatic, nucleoside-induced hyperlactatemia. METHODS: Total DNA was extracted from blood cells. A nuclear gene and a mitochondrial gene were quantified by real-time polymerase chain reaction. Three groups were studied: 24 controls not infected with the human immunodeficiency virus (HIV), 47 HIV-infected asymptomatic patients who had never been treated with antiretroviral drugs, and 8 HIV-infected patients who were receiving antiretroviral drugs and had symptomatic hyperlactatemia. The patients in the last group were studied longitudinally before, during, and after antiretroviral therapy. RESULTS: Symptomatic hyperlactatemia was associated with marked reductions in the ratios of mitochondrial to nuclear DNA, which, during therapy, averaged 68 percent lower than those of non-HIV-infected controls and 43 percent lower than those of HIV infected asymptomatic patients never treated with antiretroviral drugs. After the discontinuation of antiretroviral therapy, there was a statistically significant increase in the ratio of mitochondrial to nuclear DNA (P=0.02). In the patients followed longitudinally, the decline in mitochondrial DNA preceded the increase in venous lactate levels. CONCLUSIONS: Mitochondrial DNA levels are significantly decreased in patients with symptomatic, nucleoside-related hyperlactatemia, an effect that resolves on the discontinuation of therapy. PMID- 11893793 TI - Image in clinical medicine. Hypercarotenemia. PMID- 11893794 TI - Outcomes of reference pricing for angiotensin-converting-enzyme inhibitors. AB - BACKGROUND: In January 1997, reference pricing for angiotensin-converting-enzyme (ACE) inhibitors for patients 65 years of age or older was introduced in British Columbia, Canada. For medications within a specific class, insurance covers the cost up to the reference price, and patients pay the extra cost of more expensive medications. Although reference pricing may reduce the costs of prescription drugs, there is concern that patients may switch to less effective medications or stop treatment. METHODS: We analyzed data from the Ministry of Health on all 37,362 residents of British Columbia who were 65 or older and were enrolled in the provincial health insurance program, received ACE inhibitors priced higher than the reference price of $27 a month in 1996, and were potentially affected by the new policy. We identified 5353 residents who switched to an ACE inhibitor not subject to cost sharing during the first six months and compared them with 27,938 residents who received only ACE inhibitors subject to cost sharing. RESULTS: Reference pricing for ACE inhibitors was not associated with changes in the rates of visits to physicians, hospitalizations, admissions to long-term care facilities, or mortality. The probability of stopping antihypertensive therapy decreased as compared with the probability before the change in policy (relative risk, 0.76; 95 percent confidence interval, 0.65 to 0.89). Eighteen percent of patients who had been prescribed ACE inhibitors subject to cost sharing switched to lower-priced alternatives. As compared with patients who did not switch, those who did had a moderate transitory increase in the rates of visits to physicians (rate ratio, 1.11; 95 percent confidence interval, 1.07 to 1.15) and hospital admissions through the emergency room (rate ratio, 1.19; 95 percent confidence interval, 0.99 to 1.42) during the two months after switching, but not subsequently. CONCLUSIONS: We found little evidence that when reference pricing for ACE inhibitors was introduced in British Columbia, patients stopped treatment for hypertension or that health care utilization and costs increased. PMID- 11893795 TI - Compliance among pharmacies in California with a prescription-drug discount program for Medicare beneficiaries. AB - BACKGROUND: Several states have developed prescription-drug discount programs for Medicare beneficiaries. In California, Senate Bill 393, enacted in 1999, requires pharmacies participating in the state Medicaid program (Medi-Cal) to charge customers who present a Medicare card amounts based on Medi-Cal rates. Because Medicare beneficiaries may not be accustomed to presenting their Medicare cards at pharmacies, we assessed the compliance of pharmacies with Senate Bill 393. METHODS: Fifteen Medicare beneficiaries who received special training and acted as "standardized patients" visited a random sample of pharmacies in the San Francisco Bay area and Los Angeles County in April and May 2001. According to a script, they asked for the prices of three commonly prescribed drugs: rofecoxib, sertraline, and atorvastatin. The script enabled us to determine whether and when, during their interactions with pharmacists or salespeople, the discounts specified in Senate Bill 393 were offered. Pharmacies at which the appropriate discounts were offered were considered compliant. RESULTS: The patients completed visits to 494 pharmacies. Seventy-five percent of the pharmacies complied with the prescription-drug discount program; at only 45 percent, however, was the discount offered before it was specifically requested. The discount was offered at 91 percent of pharmacies that were part of a chain, as compared with 58 percent of independent pharmacies (P<0.001). Compliance was higher in the San Francisco Bay area than in Los Angeles County (84 percent vs. 72 percent, P=0.004) and was higher in high-income than low-income neighborhoods (81 percent vs. 69 percent, P=0.002). A Medicare beneficiary taking all three drugs would have saved an average of $55.70 per month as compared with retail prices (a savings of 20 percent). CONCLUSIONS: Discounts required under California's prescription-drug discount program for Medicare beneficiaries offer substantial savings. Many patients, however, especially those who use independent pharmacies or who live in low-income neighborhoods, may not receive the discounts. PMID- 11893796 TI - Clinical practice. Barrett's Esophagus. PMID- 11893797 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 8-2002. A 56-year-old woman with a persistent left-sided pleural effusion. PMID- 11893798 TI - Survival of the fittest--more evidence. PMID- 11893799 TI - Childhood obesity and a diabetes epidemic. PMID- 11893800 TI - Controlling spending for prescription drugs. PMID- 11893801 TI - A new element in the mechanism of asthma. PMID- 11893802 TI - Surgery for emphysema. PMID- 11893803 TI - Radiotherapy for breast cancer. PMID- 11893804 TI - Immunology series: vaccines. PMID- 11893805 TI - Persistent pulmonary hypertension of the newborn. PMID- 11893806 TI - Case 32-2001: interstitial pneumonitis and rheumatoid arthritis. PMID- 11893807 TI - Chronic thromboembolic pulmonary hypertension. PMID- 11893808 TI - Nosocomial spread of linezolid-resistant, vancomycin-resistant Enterococcus faecium. PMID- 11893809 TI - Exposing Mr Butts' tricks of the trade. Introduction. PMID- 11893810 TI - Marketing to America's youth: evidence from corporate documents. AB - OBJECTIVE: To evaluate the claim that the tobacco industry does not market its products to youth. DESIGN: The data for this study come from tobacco industry documents collected from the tobacco industry's document websites, presently linked at http://www.tobaccoarchives.com. The websites were searched using "request for production" (RFP) codes, specified keyword searches, and serendipitous terms identified in document citations found with RFP and keyword searches. RESULTS: Industry documents show that the cigarette manufacturers carefully monitored the smoking habits of teenagers over the past several decades. Candid quotes from industry executives refer to youth as a source of sales and as fundamental to the survival of the tobacco industry. The documents reveal that the features of cigarette brands (that is, use of filters, low tar, bland taste, etc), packaging (that is, size, colour and design), and advertising (that is, media placements and themes and imagery) were developed specifically to appeal to new smokers (that is, teenagers). Evidence also indicates that relevant youth oriented marketing documents may have been destroyed and that the language used in some of the more recent documents may have been sanitised to cover up efforts to market to youth. CONCLUSIONS: The tobacco industry's internal documents reveal an undeniable interest in marketing cigarettes to underage smokers. The industry's marketing approaches run counter to and predicate methods for tobacco prevention: (1) keep the price of the product high; (2) keep product placements and advertising away from schools and other areas with a high volume of youth traffic; (3) make cigarette advertising (that is, themes and visual images) unappealing to youth; (4) make product packaging unappealing to youth; and (5) design the product so it is not easy to inhale. PMID- 11893811 TI - The dark side of marketing seemingly "Light" cigarettes: successful images and failed fact. AB - OBJECTIVE: To understand the development, intent, and consequences of US tobacco industry advertising for low machine yield cigarettes. METHODS: Analysis of trade sources and internal US tobacco company documents now available on various web sites created by corporations, litigation, or public health bodies. RESULTS: When introducing low yield products, cigarette manufacturers were concerned about maintaining products with acceptable taste/flavour and feared consumers might become weaned from smoking. Several tactics were employed by cigarette manufacturers, leading consumers to perceive filtered and low machine yield brands as safer relative to other brands. Tactics include using cosmetic (that is, ineffective) filters, loosening filters over time, using medicinal menthol, using high tech imagery, using virtuous brand names and descriptors, adding a virtuous variant to a brand's product line, and generating misleading data on tar and nicotine yields. CONCLUSIONS: Advertisements of filtered and low tar cigarettes were intended to reassure smokers concerned about the health risks of smoking, and to present the respective products as an alternative to quitting. Promotional efforts were successful in getting smokers to adopt filtered and low yield cigarette brands. Corporate documents demonstrate that cigarette manufacturers recognised the inherent deceptiveness of cigarette brands described as "Light"or "Ultra-Light" because of low machine measured yields. PMID- 11893812 TI - How cigarette design can affect youth initiation into smoking: Camel cigarettes 1983-93. AB - CONTEXT: Internal industry documents may shed light on how cigarettes are designed to promote youth smoking. OBJECTIVE: To determine changes in the design of Camel cigarettes in the period surrounding the "Smooth Character" advertising campaign and to assess the impact of these changes on youth smoking. DATA SOURCES: Internal documents made available through the document website maintained by RJ Reynolds, manufacturer of Camel cigarettes. STUDY SELECTION: Electronic searches using keywords to identify relevant data. DATA EXTRACTION: A web based index search of documents targeting "smoothness" or "harshness" and "younger adult smokers" ("YAS") or "first usual brand younger adult smokers" ("FUBYAS") in the 10 year period surrounding the introduction of the "Smooth Character" campaign was used to identify Camel related product design research projects. A snowball methodology was used: initial documents were identified by focusing on key words, codes, researchers, committees, meetings, and gaps in overall chronology; a second set of documents was culled from these initial documents, and so on. DATA SYNTHESIS: Product design research led to the introduction of redesigned Camel cigarettes targeted to younger adult males coinciding with the "Smooth Character" campaign. Further refinements in Camel cigarettes during the following five year period continued to emphasise the smoothness of the cigarette, utilising additives and blends which reduced throat irritation but increased or retained nicotine impact. CONCLUSIONS: Industry competition for market share among younger adult smokers may have contributed to the reversal of a decline in youth smoking rates during the late 1980s through development of products which were more appealing to youth smokers and which aided in initiation by reducing harshness and irritation. PMID- 11893814 TI - Cigarette filter ventilation is a defective design because of misleading taste, bigger puffs, and blocked vents. AB - OBJECTIVE: To review tobacco industry documents on filter ventilation in light of published studies and to explore the role of filter ventilation in the design of cigarettes that deliver higher smoke yields to smokers than would be expected from standard machine smoked tests (Federal Trade Commission (FTC), International Organization for Standardization (ISO)). DATA SOURCES: Searched from November 1999 to November 2000 internet databases of industry documents (www.pmdocs.com, www.rjrtdocs.com, www.lorillarddocs.com, www.bw.aalatg.com, www.cdc.gov/tobacco/industrydocs, www.tobaccodocuments.org, www.tobaccopapers.org, www.hlth.gov.bc.ca/Guildford, www.cctc.ca/ncth/Guildford, www.cctc.ca/ncth/Guildford2) for documents related to filter ventilation. Documents found dated from 1955 through 1994. STUDY SELECTION: Those documents judged to contain the most relevant information or data on filter ventilation related to cigarette taste and compensatory smoking, while also trying to avoid redundancy from various documents deriving from the same underlying data. DATA SYNTHESIS: Filter ventilation is a crucial design feature creating three main problems for lower tar cigarettes as measured by official smoking machine testing. Firstly, it misleadingly makes cigarettes taste lighter and milder, and, therefore, they appear less dangerous to smokers. Secondly, it promotes compensation mainly by facilitating the taking of larger puffs. Thirdly, for very heavily ventilated cigarettes (that is, > 65% filter air dilution), behavioural blocking of vents with lips or fingers is an additional contributor to compensatory smoking. These three effects are found in industry research as well as published research. CONCLUSIONS: Filter ventilation is a dangerous, defective technology that should be abandoned in less hazardous nicotine delivery systems. Health interested groups should test cigarettes in a way that reflects compensatory smoking. Lower tar (vented filter) cigarettes should be actively countermarketed. PMID- 11893815 TI - Cigarettes with defective filters marketed for 40 years: what Philip Morris never told smokers. AB - BACKGROUND: More than 90% of the cigarettes sold worldwide have a filter. Nearly all filters consist of a rod of numerous ( > 12 000) plastic-like cellulose acetate fibres. During high speed cigarette manufacturing procedures, fragments of cellulose acetate that form the mouthpiece of a filter rod become separated from the filter at the end face. The cut surface of the filter of nearly all cigarettes has these fragments. In smoking a cigarette in the usual manner, some of these fragments are released during puffing. In addition to the cellulose acetate fragments, carbon particles are released also from some cigarette brands that have a charcoal filter. Cigarettes with filters that release cellulose acetate or carbon particles during normal smoking conditions are defective. OBJECTIVE: Specific goals were to review systematically the writings of tobacco companies to: (a) identify papers that would document the existence of defective filters; (b) characterise the extent of the defect; (c) establish when the defect became known; (d) determine whether the defect exists on cigarettes marketed currently; (e) assess the prevalence of the defect on cigarettes manufactured by different companies; (f) define whether the knowledge of the defect had been withheld by the tobacco company as confidential and not disclosed publicly; and (g) ascertain the feasibility of correcting or preventing the defect. METHODS: Document searches utilised databases of the scientific literature, medical journals, chemical abstracts, US Patents, Tobacco Abstracts, papers presented at tobacco meetings and court documents. RESULTS: Sixty one documents of Philip Morris, Inc were selected for study because they disclosed specifically the "fall out" of cellulose acetate filter fibres and, for cigarettes with charcoal filters, carbon particles from cigarette filters. The term "fall-out" was defined in 1985 laboratory protocols of Philip Morris, Inc. as "loose fibers (or particles) that are drawn out of the filter during puffing of the cigarette". As early as 1957, the health concern of inhaling cellulose acetate fibres released from cigarette filters was addressed by Philip Morris, Inc. A 1962 document reported the results of laboratory tests conducted by Phillip Morris, Inc that compared the "fall-out" of cellulose acetate fibres from the filters of their cigarettes (Marlboro) and cigarettes of their competitor (Liggett & Meyers). A 1997 overview by Phillip Morris of documents addressing the "fallout of carbon particles and cellulose acetate fibers from filters" stated that they were "essentially routine reports" of cigarette filter assays, and referenced a "Filter Fallout" memo written in 1961-more than 40 years ago. Most likely these tests are being conducted presently as illustrated by a 1999 report that details the revisions of the "fall-out" protocol of Phillip Morris, Inc and reports the results of tests that measured the discharge of cellulose acetate fibres and silica gel from beta cigarettes with a new type of filter. Our analysis of the "fall-out" tests results presented in the 61 "fall-out" documents showed that filter fibres and carbon particles were discharged from the filters of all types of cigarettes tested. These cigarette types (n = 130) included both coded cigarettes and popular brand name cigarettes. No publications were found in the scientific literature of the "fall-out" studies. Thus, the results of the "fall out" studies are thought to have been withheld as confidential to Philip Morris, Inc. We have identified also other companies that have tested recently cigarettes for defective filters. In addition, our searches have shown that simple, expedient, and inexpensive technologies for decontaminating cigarette filters of loose cellulose acetate fibres and particles from the cut surface of the filter have been developed and described in 1997 and 1998 US patents. What is more important is that these patents also define methods for preventing or reducing the broken plastic-like fibres that arise during cigarette making. Many US patents (n = 607; 1957 to 2001) have been awarded for cigarette filters. Some of these inventions describe novel materials and unique filtration schemes that would eliminate or minimise the discharge of filter materials into mainstream smoke. CONCLUSIONS: We have shown that: (a) the filter of today's cigarette is defective; (b) Philip Morris, Inc has known of this filter defect for more than 40 years; (c) the existence of this filter defect has been confirmed by others in independent studies; (d) many methods exist to prevent and correct the filter defect, but have not been implemented; and (e) results of investigations substantiating defective filters have been concealed from the smoker and the health community. The tobacco industry has been negligent in not performing toxicological examinations and other studies to assess the human health risks associated with regularly ingesting and inhaling non-degradable, toxin coated cellulose acetate fragments and carbon microparticles and possibly other components that are released from conventional cigarette filters during normal smoking. The rationale for harm assessment is supported by the results of consumer surveys that have shown that the ingestion or inhalation of cigarette filter fibres are a health concern to nearly all smokers. PMID- 11893816 TI - Tax, price and cigarette smoking: evidence from the tobacco documents and implications for tobacco company marketing strategies. AB - OBJECTIVE: To examine tobacco company documents to determine what the companies knew about the impact of cigarette prices on smoking among youth, young adults, and adults, and to evaluate how this understanding affected their pricing and price related marketing strategies. METHODS: Data for this study come from tobacco industry documents contained in the Youth and Marketing database created by the Roswell Park Cancer Institute and available through http:// roswell.tobaccodocuments.org, supplemented with documents obtained from http://www.tobaccodocuments.org. RESULTS: Tobacco company documents provide clear evidence on the impact of cigarette prices on cigarette smoking, describing how tax related and other price increases lead to significant reductions in smoking, particularly among young persons. This information was very important in developing the industry's pricing strategies, including the development of lower price branded generics and the pass through of cigarette excise tax increases, and in developing a variety of price related marketing efforts, including multi pack discounts, couponing, and others. CONCLUSIONS: Pricing and price related promotions are among the most important marketing tools employed by tobacco companies. Future tobacco control efforts that aim to raise prices and limit price related marketing efforts are likely to be important in achieving reductions in tobacco use and the public health toll caused by tobacco. PMID- 11893817 TI - The cigarette pack as image: new evidence from tobacco industry documents. AB - OBJECTIVES: To gain an understanding of the role of pack design in tobacco marketing. METHODS: A search of tobacco company document sites using a list of specified search terms was undertaken during November 2000 to July 2001. RESULTS: Documents show that, especially in the context of tighter restrictions on conventional avenues for tobacco marketing, tobacco companies view cigarette packaging as an integral component of marketing strategy and a vehicle for (a) creating significant in-store presence at the point of purchase, and (b) communicating brand image. Market testing results indicate that such imagery is so strong as to influence smoker's taste ratings of the same cigarettes when packaged differently. Documents also reveal the careful balancing act that companies have employed in using pack design and colour to communicate the impression of lower tar or milder cigarettes, while preserving perceived taste and "satisfaction". Systematic and extensive research is carried out by tobacco companies to ensure that cigarette packaging appeals to selected target groups, including young adults and women. CONCLUSIONS: Cigarette pack design is an important communication device for cigarette brands and acts as an advertising medium. Many smokers are misled by pack design into thinking that cigarettes may be "safer". There is a need to consider regulation of cigarette packaging. PMID- 11893818 TI - How the tobacco industry built its relationship with Hollywood. AB - OBJECTIVE: To describe the development of the relationship between the tobacco industry and the entertainment industry. METHODS: Review of previously secret tobacco industry documents available on the internet. RESULTS: Both the entertainment and tobacco industries recognised the high value of promotion of tobacco through entertainment media. The 1980s saw undertakings by four tobacco companies, Philip Morris, RJ Reynolds (RJR), American Tobacco Company, and Brown and Williamson to place their products in movies. RJR and Philip Morris also worked to place products on television at the beginning of the decade. Each company hired aggressive product placement firms to represent its interests in Hollywood. These firms placed products and tobacco signage in positive situations that would encourage viewers to use tobacco and kept brands from being used in negative situations. At least one of the companies, RJR, undertook an extensive campaign to hook Hollywood on tobacco by providing free cigarettes to actors on a monthly basis. Efforts were also made to place favourable articles relating to product use by actors in national print media and to encourage professional photographers to take pictures of actors smoking specific brands. The cigar industry started developing connections with the entertainment industry beginning in the 1980s and paid product placements were made in both movies and on television. This effort did not always require money payments from the tobacco industry to the entertainment industry, suggesting that simply looking for cash payoffs may miss other important ties between the tobacco and entertainment industries. CONCLUSIONS: The tobacco industry understood the value of placing and encouraging tobacco use in films, and how to do it. While the industry claims to have ended this practice, smoking in motion pictures increased throughout the 1990s and remains a public health problem. PMID- 11893819 TI - Tobacco related bar promotions: insights from tobacco industry documents. AB - OBJECTIVES: To examine the tobacco industry's use of bar promotions, including their target groups, objectives, strategies, techniques, and results. DESIGN: Over 2000 tobacco industry documents available as a result of the Master Settlement Agreement were reviewed on the internet at several key web sites using keyword searches that included "bar", "night", "pub", "party", and "club". The majority of the documents deal with the US market, with a minor emphasis on Canadian and overseas markets. RESULTS: The documents indicate that bar promotions are important for creating and maintaining brand image, and are generally targeted at a young adult audience. Several measures of the success of these promotions are used, including number of individuals exposed to the promotion, number of promotional items given away, and increased sales of a particular brand during and after the promotion. CONCLUSION: Bar promotions position cigarettes as being part of a glamorous lifestyle that includes attendance at nightclubs and bars, and appear to be highly successful in increasing sales of particular brands. PMID- 11893820 TI - Tobacco Institute lobbying at the state and local levels of government in the 1990s. AB - OBJECTIVE: To describe variation in Tobacco Institute (TI) lobbying expenditures across states and test whether these expenditures vary in relationship to measures of tobacco control activity at the state level. INDEPENDENT VARIABLE: Data for this study came from the TI's State Activities Division (SAD) annual budgets for the years 1991-97, excluding 1993. These data include budgetary information pertaining to state and local lobbying activity and special projects reported by state. DEPENDENT VARIABLES: The following measures of state tobacco control activity during the period 1991 to 1997 were considered: (1) American Stop Smoking Intervention Study (ASSIST) funding; (2) voter initiatives to raise cigarette taxes; (3) cigarette excise tax level; (4) workplace smoking restrictions; (5) the intensification of smoke-free air laws covering private worksites, government worksites, and restaurants; (6) the intensification of strength of sales to minors laws; (7) the intensification of strength of laws that punish minors for possessing, purchasing, and/or using cigarettes; (8) state status as a major grower of tobacco; (9) partisan control of state government, 1996; and (10) an overall composite index reflecting a state's strength of tobacco control, combining cigarette prices with workplace and home smoking bans. RESULTS: The overall annual budget for the TI declined steadily during the 1990s, from $47.7 million in 1991 to $28.1 million by 1996. The proportion of the TI's budget allocated to the SAD remained relatively stable at about 30%. TI expenditures for lobbyists were highest in California where tobacco control activity has been strong for the past decade. We found significant associations between TI SAD expenditures and cigarette excise tax levels, the status of a state as a recipient of federal ASSIST funds, and changes in the strength of statewide laws that penalise minors for possessing, purchasing, and/or using cigarettes. We found little or no association between state and local lobbying budgets of the TI and changes in statewide smoke-free air laws, although we did find evidence of TI special project expenditures earmarked to specific states and localities to resist clean indoor air legislation/regulations (that is, Maryland and New York City). We found no significant correlation between TI lobbying expenditures and sales to minors' laws, status as a major producer of tobacco, or partisan control of state government. CONCLUSIONS: The findings from this study support the hypothesis that in the 1990s tobacco control activities such as raising cigarette excise taxes and participation in ASSIST attracted TI resources to undermine these efforts. PMID- 11893821 TI - Failed promises of the cigarette industry and its effect on consumer misperceptions about the health risks of smoking. AB - BACKGROUND: In January 1954, US tobacco manufacturers jointly sponsored an advocacy advertisement entitled "A Frank Statement to Cigarette Smokers" which appeared in 448 newspapers in 258 cities reaching an estimated 43 245 000 Americans. The advertisement questioned research findings implicating smoking as a cause of cancer, promised consumers that their cigarettes were safe, and pledged to support impartial research to investigate allegations that smoking was harmful to human health. OBJECTIVE: To examine (1) the extent to which cigarette companies fulfilled the promises made to consumers in the 1954 "Frank Statement", and (2) the effect of these promises on consumer knowledge, beliefs, and smoking practices. METHODS: This study reviews statements made since 1954 by the tobacco companies individually and collectively through the Tobacco Institute and Tobacco Industry Research Committee/Council for Tobacco Research on the subject of smoking as a cause disease, and the industry's pledge to support and disclose the results of impartial research on smoking and health. Many of the industry documents evaluated in this study were obtained from a collection consisting of 116 documents entitled the "Statement of Defendants' Misrepresentations" prepared by attorneys representing the state of Connecticut in the Medicaid litigation against the tobacco industry in 1998. In addition, we searched for corroborating material from tobacco industry documents collected from the tobacco industry's document websites. In order to contrast industry statements on smoking and health with what smokers' actually believed about smoking we reviewed reports of public polling data on smokers' knowledge and beliefs about smoking and disease gathered from tobacco industry sources and from surveys conducted by public health researchers. RESULTS: Analysis of public statements issued by the tobacco industry sources over the past five decades shows that the companies maintained the stance that smoking had not been proven to be injurious to health through 1999. The public statements of the tobacco industry are in sharp contrast to the private views expressed by many of their own scientists. The tobacco documents reveal that many scientists within the tobacco industry acknowledged as early as the 1950s that cigarette smoking was unsafe. The sincerity of the industry's promise to support research to find out if smoking was harmful to health and to disclose information about the health effects of smoking can also be questioned based upon the industry's own documents which reveal: (1) scepticism about the scientific value of the smoking and health research program established by the industry; and (2) evidence that research findings implicating smoking as a health problem were often not published or disclosed outside the industry. Industry documents also show that the companies knew that their own customers were misinformed about smoking and health issues. CONCLUSION: It is clear that the cigarette companies failed to fulfill the promises made to consumers in the 1954 "Frank Statement" advertisement. The failure of cigarette manufacturers to honour these promises has resulted in a public that even today remains misinformed about the health risks of smoking. PMID- 11893822 TI - Autoantibodies to cardiolipin and beta-2-glycoprotein-I in coronary artery disease patients with and without hypertension. AB - BACKGROUND: Autoantibodies to cardiolipin (aCL) and beta2-glycoprotein-I (beta2GPI) are considered proatherogenic. Elevated levels of both antibodies have been reported in hypertension. Nonetheless, there are no data regarding an association between these autoantibodies and hypertension in coronary artery disease. METHODS: The levels of aCL and anti-beta2GPI antibodies were measured in patients having coronary artery disease with (n = 82) or without (n = 36) hypertension, in association with other major risk factors for coronary artery disease. RESULTS: The levels of aCL and anti-beta2GPI antibodies were (OD at 405 nm) 0.23 +/- 0.14 and 0.22 +/- 0.12 in the normal blood pressure group, as opposed to 0.24 +/- 0.12 and 0.20 +/- 0.12 in the hypertensive group, respectively (p = 0.67; 0.42). No significant difference in either antibody levels was found between hypertensive patients with normal and abnormal blood pressure measurements. The presence of additional risk factors did not affect antibody levels in normotensive patients. However, in the hypertensive group, the presence of smoking was associated with significantly decreased anti-beta2GPI antibody levels, whereas no change was found in aCL. Further, patients who had hypertension, smoking and hypercholesterolemia, had significantly decreased anti beta2GPI antibody levels compared with patients without any of these risk factors (0.13 +/- 0.04 versus 0.23 +/- 0.13, respectively; p = 0.02). CONCLUSION: Hypertension is not associated with modification of aCL and anti-beta2GPI antibody levels in coronary artery disease patients. However, there are elevated anti-beta2GPI antibody levels in patients without conventional risk factors compared with patients with these risk factors. PMID- 11893823 TI - Outcome and cost-effectiveness of cardiopulmonary resuscitation after in-hospital cardiac arrest in octogenarians. AB - CONTEXT: Octogenarians are the fastest growing segment of the population and little is known about the results of cardiopulmonary resuscitation (CPR) after in hospital cardiac arrest in this population. OBJECTIVE: We sought to investigate the clinical benefit and cost-effectiveness of CPR after in-hospital cardiac arrest in octogenarians. MAIN OUTCOME MEASURE: Years of life saved. DESIGN: Effectiveness data were obtained from a review of 91,372 hospital discharges from January 1st, 1993 until June 30th, 1996. Cardiac arrest was reported in 956 patients. The study group consisted of 474 patients > or = 80 years old. CPR costs included equipment and training, physician and nursing time and medications. Post-CPR expenses included in-hospital true cost, repeat hospitalizations, physician office visits, nursing home, rehabilitation, and chronic care hospital costs. Life expectancy of the patients who were still alive at the end of the study was estimated from census data. A utility of 0.8 was used to calculate quality-adjusted-life years saved (QALYS). We used a societal perspective for analysis. RESULTS: The study population was 86 +/- 4.8 years old (range 80-103), and 42% were male. Fifty-four patients (11%) were discharged alive, 35 to a chronic care facility and 19 to their home. Assuming that a cardiac arrest without CPR has 100% mortality, 12 octogenarians required treatment with CPR in order to save one life to hospital discharge. Similarly, 29 octogenarian patients with cardiac arrest have to be treated with CPR to net one long-term survivor (mean survival 21 months, with a range from 9 to 48 months). The cost-effectiveness ratio, after estimating the life expectancy of octogenarian survivors, was USD 50,412 per year of life saved, and USD 63,015 per QALYS. However, a utility of 0.5 yielded a cost of USD 100,825 per QALYS. CONCLUSION: In comparison with other life-saving strategies, CPR in octogenarians is effective. The favorable cost-effectiveness ratio is highly dependent on the patients' preference for quality rather than quantity of life, as expressed by the utility assumptions. PMID- 11893824 TI - Plasma levels of brain natriuretic peptide increase in patients with idiopathic bilateral atrial dilatation. AB - Idiopathic bilateral atrial dilatation (IBAD) is an extremely rare anomaly and is usually associated with atrial fibrillation. Plasma levels of atrial natriuretic peptide (ANP) have been shown to increase in patients with atrial fibrillation. However, secretion of ANP and brain natriuretic peptide (BNP) in patients with IBAD remains unclear. We investigated the clinical features of 9 patients with IBAD and 16 age- and sex-matched patients with lone atrial fibrillation (LAF). Plasma levels of ANP and BNP were measured, and echocardiographic parameters were followed. Left (LAV) and right atrial volumes (RAV) were significantly higher in patients with IBAD than in patients with LAF (both p < 0.01). There were no differences between patients with IBAD and LAF in other echocardiographic parameters. The percent increases in LAV and RAV in patients with IBAD exceeded those in patients with LAF (both p < 0.01). Plasma levels of BNP and the BNP/ANP ratios in patients with IBAD were significantly higher than those in patients with LAF (both p < 0.01), but there was no significant difference in plasma levels of ANP. Regarding the clinical course of the patients with IBAD compared with those with LAF, the atrial volume increased gradually, and plasma levels of BNP were significantly higher. These findings suggested that IBAD was not only influenced by long-term atrial fibrillation, but also by subclinical left ventricular dysfunction. PMID- 11893825 TI - Gender gap in coronary artery disease: comparison of the extent, severity and risk factors in men and women aged 45-65 years. AB - This retrospective study aimed to characterize coronary artery disease (CAD) and its risk factors among relatively young women, as compared to men in a similar age group. Confirmed cases of CAD were compared regarding their medical background, performance and outcome of coronary artery procedures, physical profile and lifestyle information. The study population included 179 women and 270 men aged 45-65 years who were hospitalized during the study period 1990-1995 in the Hadassah Medical Centers. Significantly more women presented with histories of prior myocardial infarction and a higher number of vessels occluded by 80% or more and required percutaneous transluminal coronary angioplasty for 3 or more arteries, and the women had a higher incidence of risk factors such as diabetes, hypertension and hypercholesterolemia than their male counterparts. PMID- 11893826 TI - A new guidewire technique for difficult cardiac catheterization. AB - PURPOSE: To propose a simple guidewire technique to resolve the problem of difficulty in advancing catheters or guidewires during vascular and cardiac catheterization. METHODS: A 0.035-inch J-tip guidewire was reshaped manually at the distal 7- to 8-cm portion, including the flexible part and 2-3 cm of the stiff part, into zigzag shape. With this modification, the guidewire became much more trackable and could be advanced to the right position. RESULTS: In the past 2 years, this 'zigzag wire technique' was applied in cardiac catheterization with high success rates for cases with difficulty in advancing either catheters or guidewires. This wire technique was applied in peripheral arterial tortuosity or stenosis (18 cases, with success in 17), aortic aneurysm (successful in all 7 cases), aortic stenosis (11 cases, successful in 8), and dilated right heart chambers (successful in all 6 cases). No complications resulted from the use of this guidewire technique. CONCLUSION: This simple method of guidewire modification would be very helpful in difficult vascular and cardiac catheterization when difficulty in advancing catheters or wires was encountered. PMID- 11893827 TI - Angiotensin-converting enzyme insertion/deletion polymorphism does not influence the restenosis rate after coronary stent implantation. AB - BACKGROUND: Experimental studies have shown an activation of the angiotensin converting enzyme (ACE) system as a response to endothelial injury. Recent publications have elucidated the hypothesis that the ACE gene polymorphism may influence the level of late luminal loss after coronary stent implantation. It is still unclear whether the polymorphism of the angiotensin gene is a major predictor of the extent of neointimal hyperplasia. In this multicenter study, we therefore tested the relationship between the ACE gene polymorphism and the restenosis rate after coronary stent implantation. METHODS: As a substudy of the optimization with intracoronary ultrasound (ICUS) to reduce stent restenosis (OPTICUS) study, we analyzed ACE serum levels and the ACE gene polymorphism in 154 patients at 9 different centers. All patients underwent elective coronary stent implantation in a stenosis of a major coronary vessel. Balloon inflations were repeated until a satisfactory result was achieved in on-line quantitative coronary angiography or ICUS fulfilling the OPTICUS study criteria. After follow up of 6 months, all patients underwent reangiography under identical projections as the baseline procedure. A blinded quantitative analysis of the initial procedure as well as the follow-up examinations were performed by an independent core laboratory. ACE gene polymorphism and ACE serum activity were measured at the 6-month follow-up in a double-blinded setting. RESULTS: With respect to the ACE gene polymorphism, there were three subgroups: DD genotype (48 patients), ID (83 patients) and II (23 patients). The subgroups did not differ in regard to age, gender, extent of coronary artery disease, stenosis length, initial degree of stenosis or degree of stenosis after stent implantation. In all, 39 patients (25.3%) had significant restenosis: 12 DD patients (25.0%), 18 ID patients (21.7%) and 9 II patients (39.1%) (odds ratio 2.164, 95% confidence interval 0.853-5.493). We obtained the following results for ACE serum levels: 0.53 micromol/l/s in the DD subgroup, 0.29 micromol/l/s in the ID subgroup and 0.09 micromol/l/s in the II subgroup (p < 0.001). Multivariate logistic regression analysis of the influence of ACE gene polymorphism on the restenosis rate after coronary stent implantation adjusted for lesion length (>12 mm), ACE inhibitor or hydroxymethylglutaryl coenzyme A reductase (CSE) inhibitor treatment, age, male gender, diabetes mellitus, hypertension, high cholesterol, family history, smoking and three-vessel disease did not uncover any statistic significance. CONCLUSIONS: In contrast to other study groups, we were unable to disclose that the DD polymorphism of the ACE gene was associated with a higher rate of restenosis after coronary stent implantation in this multicenter study. In addition, patients with higher ACE serum levels did not show a higher restenosis rate in this trial. We conclude that the pathogenesis of restenosis is a multifactorial process involving various genetic and nongenetic factors. PMID- 11893828 TI - Dipyridamole myocardial perfusion tomography in patients with severe aortic stenosis. AB - Patients with aortic stenosis (AS) may have classic angina pectoris. The safety of exercise testing in adults with AS is controversial and, in fact, exercise testing in such patients is considered to be contraindicated especially in severe aortic stenosis (SAS). Furthermore, exercise testing has low specificity in uncovering coronary artery disease (CAD) in patients with AS, because the baseline ECG is frequently abnormal. We wished to assess the safety and diagnostic accuracy of dipyridamole stress myocardial perfusion tomography (DMPT) in the detection of CAD in patients with SAS. METHODS: The study included 30 patients with SAS (mean aortic valve area 0.57 +/- 0.09 cm(2)). All patients underwent dipyridamole myocardial perfusion scintigraphy (SPECT), coronary arteriography and catheterization, as well as Doppler echocardiography. Myocardial perfusion tomography was applied with (99m)Tc hexakis-2 methoxyisobutyl isonitrile (MIBI) by a single day rest-dipyridamole infusion protocol. Hemodynamic, electrocardiographic and clinical responses were compared with those of 50 control patients without AS. RESULTS: Hemodynamic responses during dipyridamole stress tests demonstrated no significant differences between the controls and the AS patients in the following parameters: systolic blood pressure, heart rate, rate-pressure product or incidence of headache, chest pain, dyspnea, flushing and dizziness. A reversible perfusion defect was observed in 10 patients with DMPT. The existence of coronary lesions was determined by coronary arteriography in 8 of 10 patients (sensitivity 100%, specificity 91%). CONCLUSION: The results showed that DMPT is well tolerated, even by patients with SAS and is of high diagnostic value in assessing CAD. PMID- 11893830 TI - Arrhythmogenic right ventricular dysplasia masquerading as peripartum cardiomyopathy with atrial flutter, advanced atrioventricular block and embolic stroke. PMID- 11893829 TI - Long-term follow-up with stress echocardiograms of patients with Kawasaki's disease. AB - Patients with a history of Kawasaki's disease (KD), particularly those not treated with intravenous gamma-globulin, are at risk of coronary artery aneurysms and later obstruction. Twenty-eight patients with a history of KD (4 had coronary artery aneurysms) were examined with stress echocardiograms. Fourteen patients received gamma-globulin < or =10 days of the onset, 8 patients received gamma globulin >10 days and 6 received no gamma-globulin. The mean age at diagnosis was 7.2 +/- 4.1 years; the median follow-up was 8.0 +/- 7.4 years. All tests were negative. Using a binomial model, a power of 0.80, a sensitivity of each test of 80% and assuming uniform risk, the individual rate of failure to detect was <7%. At least 640 patients in each group would be needed to detect a difference of 3.5% vs. 7.0% and 184 in each group would be needed to detect a difference of 1.5% vs. 7.5%. We conclude that the probability of an abnormal stress echo in asymptomatic patients with a history of KD is at most 7% and that a more precise determination of the risk of an abnormal stress echo in KD requires a much larger study. PMID- 11893831 TI - Sinus of Valsalva aneurysm diagnosed by transesophageal echocardiography. Case report and short review of published data. PMID- 11893832 TI - Massive pulmonary artery thrombosis, pulmonary hypertension and untreated atrial septal defect. PMID- 11893833 TI - Apolipoprotein E among Korean Alzheimer's disease patients in community-dwelling and hospitalized elderly samples. AB - This study evaluated the association of the apolipoprotein E epsilon4 allele (epsilon4) with Alzheimer's disease (AD) and investigated the relationships of epsilon4 with clinical characteristics of AD in both community-dwelling and hospitalized Korean elderly populations. The apolipoprotein E genotypes were compared between a group of 52 community-dwelling patients with AD (C-AD), a group of 52 hospitalized patients with AD (H-AD) and a group of 52 healthy controls matched to C-AD. Clinical characteristics and scores on several assessment scales were compared between C-AD and H-AD as well as between two subgroups within each AD group: those with epsilon4 and those without epsilon4. For H-AD, the assessment scales were administered again 6 months later, and the temporal course of AD was compared between the subgroups. The frequency of epsilon4 was significantly higher in both AD groups than in controls. Between C AD and H-AD, no differences were found in epsilon4 frequency, while symptoms of dementia were significantly severer in H-AD than in C-AD. Within each AD group, no differences were observed in terms of clinical characteristics and scores on the assessment scales between the subgroups. Furthermore, the subgroups of H-AD showed no difference in the rate of progression. In conclusion, there was a significant association of epsilon4 with AD. However, no relationships were found between epsilon4 and clinical characteristics in either C-AD or H-AD. Furthermore, the rate of progression did not vary with respect to the presence or absence of epsilon4 in H-AD. The severer symptoms observed in the hospitalized sample than in those at large might be explained by factors other than the impact of epsilon4. PMID- 11893834 TI - The domain-specific, stage-limited impact of the apolipoprotein E epsilon-4 allele on cognitive functions in Alzheimer's disease. AB - To examine the impact of the APOE epsilon4 allele on the cognitive functions of Alzheimer's disease (AD) patients, we administered the eight neuropsychological tests from the Consortium to Establish a Registry for Alzheimer's Disease Neuropsychological Assessment Battery to 118 Korean AD patients. The impact of the APOE epsilon4 allele was significant in the Word List Recall Test (WLRT) and the Word List Recognition Test (WLRcT) only, and its impact was confined to the very mild AD (VMAD) patients (F = 7.65, d.f. = 2, p < 0.01 for WLRT; F = 3.27, d.f. = 2, p = 0.04 for WLRcT). In the VMAD group, the performance on the two tests of the APOE-epsilon4-positive patients was poorer than that of the APOE epsilon4-negative patients. Our findings suggest that the impact of the APOE epsilon4 allele on cognitive functions in AD may be domain specific and confined to the early stage of AD. PMID- 11893835 TI - Contributions of other brain pathologies in dementia with lewy bodies. AB - The clinical picture with its pathological correlate was analysed in 16 patients fulfilling consensus criteria for dementia with Lewy bodies (DLB). The cases were part of a larger cohort (n = 200) of patients within a prospective longitudinal study of dementing disorders. Six cases exhibited not only Lewy bodies (LBs) but also other brain pathologies such as Alzheimer changes, multiple infarcts or complete and incomplete white matter infarcts. Degeneration of the nucleus basalis of Meynert and substantia nigra was also seen. The 10 cases without LBs all had Alzheimer changes. In 7 cases, these changes were combined with mainly incomplete frontal white matter infarcts. However, the degeneration of brain stem nuclei was less pronounced in these cases. Symptoms such as fluctuations in cognition, falls and episodic confusion appeared in association with arterial hypotension, which developed during the course of dementia in almost all the 16 cases. The majority of the cases were treated with neuroleptics and other potentially hypotensive medication. This study shows that multiple and different pathological features may contribute to a clinical symptom constellation as in DLB. The case study approach reveals the complexity of the clinico-pathological relationships in dementia that might otherwise be lost in the analysis of larger group data. PMID- 11893836 TI - Characteristics of two telephone screens for cognitive impairment. AB - We studied 56 subjects, 30 patients with a clinical diagnosis of Alzheimer's disease (AD) and 26 healthy controls, using two telephone screens for cognitive impairment, a self-report interview referred to as the TELE and the Telephone Interview for Cognitive Status (TICS). The sensitivity and specificity of the TELE to differentiate AD patients from healthy controls was 90.0 and 88.5% and those of the TICS were 86.7 and 88.5%, respectively. When receiver operator characteristic curves were constructed, the area under the curve for the TELE was 96.0% (SE 2.4%) and for the TICS 90.3% (SE 4.2%). Pearson's correlation between the TELE and the Mini-Mental State Examination (MMSE) was 0.87 (p < 0.0001) and between the TICS and the MMSE 0.86 (p < 0.0001). The correlation between the TELE and the sum of the boxes of the Clinical Dementia Rating scale (CDR-SB) was -0.71 (p < 0.0001) and -0.75 between the TICS and the CDR-SB (p < 0.0001). These results indicate that both screens are sensitive and specific instruments for differentiating AD patients from healthy controls and have a strong correlation with face-to-face measures of cognitive function. PMID- 11893837 TI - Cognitive impairment and mortality among nonagenarians: the Danish 1905 cohort survey. AB - Cognitive impairment has been associated with increased mortality. Most studies, however, have only included small numbers, if at all, of the very old. In a large nationwide survey of all Danes born in 1905 and still alive in 1998, where the baseline examination was conducted, we examined the impact of cognitive impairment on mortality over a 2-year period. No cognitive impairment was defined as a score of 24-30 points on the Mini Mental State Examination, mild cognitive impairment was defined as a score of 18-23 points, and severe impairment was defined as a score of 0-17 points. Cox regression analysis was applied to adjust for a number of known and suspected factors known or suspected of being associated with cognition and mortality (e.g. sociodemographic factors, sex, smoking, alcohol consumption, depressive symptoms, and physical abilities), and yielded hazard ratios (95% confidence interval) of 1.24 (1.00-1.55) for mildly impaired and 1.73 (1.37-2.20) for severely impaired Danes compared to individuals with no impairment. Cognitive impairment predicts mortality among the very old, even after controlling for most known predictors of mortality. PMID- 11893838 TI - Individual quality of life factors distinguishing low-burden and high-burden caregivers of dementia patients. AB - Dementia patient (n = 72) and caregiver characteristics and individual quality of life (IQoL) factors distinguishing low- and high-burden caregivers were evaluated. Measures included patient cognitive, functional and behavioural status, and caregiver burden, well-being, social support appraisal and IQoL. The caregivers were divided by median split into low- and high-burden groups. In the high-burden group daughters were over-represented, psychological morbidity was higher, QoL was lower, the patients were more behaviourally disturbed, and there was a trend towards more negative appraisal of informal social support. Of the many QoL factors elicited from caregivers, only 'time for self' and 'finances' differed significantly between the groups. A need for more time away from the patient is a major QoL concern for highly burdened caregivers, and a perceived lack of adequate informal support and/or financial constraints are contributory factors. PMID- 11893839 TI - Cognitive deficits and polymorphism of apolipoprotein E in Alzheimer's disease. AB - The aim of this study was to test the relationship between apolipoprotein E (ApoE) genotypes and patterns of cognitive deficits in Alzheimer's disease (AD). All subjects were diagnosed as probable AD patients on the basis of the DSM-IV and NINCDS-ADRDA criteria. Each subject was examined for (1) ApoE genotype, (2) general level of mental activity (Global Deterioration Scale and Mini-Mental State Examination) and (3) cognitive functions by means of a battery of neuropsychological tests. On the basis of ApoE genotype, patients were subdivided into two groups: the first group consisted of patients with at least one epsilon4 allele (epsilon4+ group), while the second one consisted of patients without the epsilon4 allele (epsilon4- group). Our results showed that several cognitive processes depended on the ApoE genotype. In early stages of AD, patients from the epsilon4+ group had greater deficits in delayed recall of new information. On the other hand, working memory appeared to be more impaired in the epsilon4- group of patients. Independent of the genotype, both groups showed similar impairment of learning ability without, however, deficits in remote memory. PMID- 11893840 TI - Increased serum levels of CD95 in Alzheimer's disease. AB - There is growing evidence for a role of apoptosis in Alzheimer's disease (AD). Recent findings suggest an increased susceptibility of lymphocytes to apoptosis in AD. To prove the hypothesis of systemic alterations in the apoptotic balance in AD, serum and cerebrospinal fluid levels of soluble CD95, which is known to mediate apoptosis, were measured. In the serum, AD patients exhibited significantly higher levels of CD95 than the controls (p = 0.017), suggesting an involvement of peripheral markers in AD. PMID- 11893842 TI - Effect of surgical menopause on cognitive functions. AB - To investigate the effect of estrogen deficiency on cognitive function in surgically menopausal women, a prospective study was conducted at the University Hospital in Assiut, Egypt, during the period of July 1997 to August 1999. The study included 35 women subjected to total abdominal hysterectomy and bilateral salpingo-oophorectomy for nonmalignant causes. They were subjected to cognitive assessment by Mini-Mental State Examination (MMSE), Wechsler Memory Scale (WMS) subtests, and measurement of auditory Event-Related Potentials (ERPs) and serial serum estradiol levels determination. Eighteen age- and education-, body-weight- and parity-matched control women were recruited for comparison. A significant decline in MMSE, WMS subtests (digit span, visual memory, logical memory and mental control) and prolongation of P300 of ERP latency was observed in the patient group at 3 and 6 months postoperatively. These changes were not observed in the control group. A significant correlation was found between serum estradiol level and mental control subtest score and P300 latency in patients preoperatively. Patients who had a drop of estrogen level >50% had more cognitive function decline. Rapid decline in estrogen level following surgical menopause was associated with a deleterious effect on cognitive function. Such observations may contribute to more understanding of the age-related cognitive decline in females. PMID- 11893841 TI - Effects of rivastigmine on cognitive function in dementia with lewy bodies: a randomised placebo-controlled international study using the cognitive drug research computerised assessment system. AB - This study was designed to assess the effects of rivastigmine (Exelon) on the cognitive functioning of patients suffering from dementia with Lewy bodies. This was a prospective, multi-centre, randomised, double-blind, placebo-controlled exploratory study conducted at sites in the UK, Spain and Italy. The treatment period was 20 weeks with a 3-week posttreatment follow-up. The primary outcome measures were the Cognitive Drug Research (CDR) computerised assessment system and the Neuropsychiatric Inventory. Testing was conducted prior to dosing and then again at weeks 12, 20 and 23. Analysis of the data from the 92 patients who completed the study identified a significant pattern of benefits of rivastigmine over placebo on the CDR system. These benefits were seen on tests of attention, working memory and episodic secondary memory. Taking attention for example, patients given placebo showed a significant deterioration from predosing scores at 12 and 20 weeks, whereas patients on rivastigmine performed significantly above their predosing levels. These effects were also large in magnitude, the decline under placebo at week 12 being 19%, while the improvement under rivastigmine was 23%. The clinical relevance of this 23% improvement was that it took the patients 33% towards being normal for their age on this assessment of attention. These benefits to cognitive function were accompanied by a significant improvement of the other primary outcome measure, the Neuropsychiatric Inventory. Three weeks after discontinuation of rivastigmine, most parameters of cognitive performance returned to predrug levels. PMID- 11893843 TI - [Regelsberger's intravenous oxygen therapy--an interpretation of results in practice from a biochemical and physiological point of view]. AB - Results of intravenous (i.v.) application of medical oxygen described by users are interpreted by means of biochemical and physiological knowledge. It is shown that i.v. oxygen therapy produces changes in metabolism which can also occur independently of the course of illness. Apart from the general improvement in oxygen availability, i.v. oxygen therapy causes eosinophilia, which can be valued as an increase in undetermined cellular immunological resistance. Furthermore, rheological qualities of the blood as well as diuresis are improved, the release of oxygen into the tissue is increased, and the blood pH is normalized. Compared with hyperbaric oxygen therapy, i.v. oxygen therapy seems to have less side effects. Application is less complicated, less expensive but probably of higher efficacy. This statement does not apply to issues related to diving medicine. PMID- 11893844 TI - [Distant healing and diabetes mellitus. A pilot study]. AB - BACKGROUND: The Institut fur Grenzgebiete der Psychologie und Psychohygiene, Freiburg (IGPP) in cooperation with the Abteilung Naturheilkunde, University Hospital, Zurich investigated whether Distant Healing has a beneficial effect on patients with diabetes mellitus regarding the state of the disease and quality of life. OBJECTIVE: The goal of the pilot study was to observe the progression of the disease with various medical and psychological measures and to explore which of them might be sensitive for measuring possible treatment effects. PATIENTS AND METHODS: 14 diabetic patients were observed for a period of 16 weeks. Within this time they underwent a treatment of 4 consecutive weeks (weeks 9-12) by 5 experienced and trustworthy healers each. Patients were informed about the duration of the treatment but not about the time point of its beginning. Patients and healers never met and there was no contact between researchers and patients during the study period. RESULTS: With regard to medical parameters, reduction in fructosamine level was observed during the healing period, increasing fructosamine level after the end of the healing period. Sensitivity, measured only at the beginning and at the end of the study period, decreased significantly. The other parameters showed some significant changes but there was no correlation to the Distant Healing intervention. Regarding the psychological data, only improvements were observed. CONCLUSIONS: The results indicate the possibility that a Distant Healing intervention could have certain effects on patients with diabetes mellitus. PMID- 11893845 TI - [Non-reciprocal social exchange is a health risk: a medical sociological research model]. AB - Contrary to a reductionist disease concept prevailing in molecular medicine, a systemic disease concept is emphasised in this sociopsychosomatic approach towards analysing disease as a result of disturbed social exchange among people. More precisely, violations of the norms of social reciprocity in core social roles in adult life, in particular in the work role, are assumed to trigger stressful experience with adverse long-term consequences for health. The model of effort-reward imbalance at work provides an illustrative case of nonreciprocal social exchange. It defines distinct conditions in which an imbalance between high efforts spent and low rewards received in turn is maintained by the workers. In this model social rewards are analysed in terms of money, esteem and promotion prospect including job security. The contribution summarises results from 6 international prospective and cross-sectional epidemiological investigations testing the model of effort-reward imbalance at work with regard to different health indicators (especially coronary heart disease, hypertension, depression, alcohol dependence). In all instances an elevated risk of illness is observed among those who experience non-reciprocal social exchange in terms of effort reward imbalance at work, compared to people who are free from this type of stressful experience. Results are derived from logistic regression analysis adjusting for the effects of potential confounders. In view of this evidence the medical sociological approach outlined here may be helpful in contrasting a reductionist concept of disease with a systemic concept centred around the sociopsychosomatic aspects of human health and disease. PMID- 11893848 TI - Pathophysiology of chronic urticaria. AB - Chronic urticaria includes several different subsets with distinct pathophysiologies, and with important implications for investigation and treatment. Chronic 'idiopathic' urticaria represents a special challenge, which, until recently, was not taken up by dermatological or immunological investigators. However, it has now emerged that at least 30% of patients possess histamine-releasing autoantibodies against Fc epsilon R1, or less commonly IgE itself. These autoantibodies are causative. Recent work implicates complement activation in most cases. Functional (i.e. histamine releasing) autoantibodies are specific to chronic urticaria. However, immunoreactive (non-histamine releasing) anti-Fc epsilon R1 autoantibodies can be found in sera of patients with physical urticarias and with autoimmune connective tissue and bullous diseases. The reason for the occurrence of this disease type in some individuals but not others is unclear. One possibility is the development, in genetically predisposed persons, of autoantibodies by molecular mimicry - perhaps against lipopolysaccharide of Helicobacter pylori, an organism frequently infecting the upper gastrointestinal tract of chronic urticaria patients. PMID- 11893849 TI - Treatment of rheumatoid arthritis by TNF-blocking agents. AB - Chimeric, humanized and fully human monoclonal antibodies directed against tumor necrosis factor-alpha as well as TNF receptor constructs can be administered relatively safely during long-term use for the treatment of rheumatoid arthritis (RA). Their therapeutic efficacy in patients refractory to treatment with conventional disease-modifying drugs was proven in large clinical trials and their ability to slow the progression of disease was demonstrated radiographically. The insights into the pathophysiology of RA provided by the beneficial effects of blocking proinflammatory cytokines will lead to further drug development for this destructive autoimmune disease. PMID- 11893846 TI - Validation of questionnaires from several medical fields regarding the constitution of patients. AB - INTRODUCTION: To consider or determine an individual's 'warm and cold' constitution is common part of traditional Chinese medicine (TCM), Indian medicine (Ayurveda), anthroposophic medicine and classical natural medicine. OBJECTIVE: Common psychometric characteristics of 4 questionnaires should be examined. METHODS: 110 cancer patients and 110 non-cancer patients were asked to answer these questionnaires twice 4 weeks apart. Subsequent validation of the questionnaires included psychometrical qualities of item acceptability, sensitivity to change, construct validity (homogeneity), re-test reliability and group comparison. RESULTS: The Ayurveda questionnaire was considered suitable after reducing the questions from 22 to 8 (internal construct validity kappa = 0.64). The TCM questionnaire was considered unsuitable in our model because of the absence of construct validity. The questionnaire on anthroposophic medicine was considered a suitable and reliable tool with alpha = 0.57 (Cronbach's alpha) and r = 0.61 (re-test reliability) after reducing the questions from 6 to 4. The questionnaire on sensitivity to temperatures based on classical natural medicine was considered suitable after reducing the questions from 10 to 8 (alpha = 0.58; r = 0.73). Cancer and non-cancer patients differ in only one item, i.e. in their sensitivities to cold. CONCLUSION: The questionnaires for anthroposophic medicine, Ayurveda medicine and sensitivity to temperatures are in part suitable and reliable for determining individual constitutions according to their medical philosophy. The questionnaire based on traditional Chinese medicine failed these criteria. A difference between patients with or without malignant disease was only observed for sensitivity to temperatures. The anthroposophic hypothesis that cancer either changes an individual's thermic constitution or is caused by such a change could not be confirmed in this study. PMID- 11893850 TI - cDNA cloning, biological and immunological characterization of the alkaline serine protease major allergen from Penicillium chrysogenum. AB - BACKGROUND: Penicillium chrysogenum (Penicillium notatum) is a prevalent airborne Penicillium species. A 34-kD major IgE-reacting component from P. chrysogenum has been identified as an alkaline serine protease (Pen ch 13, also known as Pen n 13 before) by immunoblot and N-terminal amino acid sequence analysis. METHODS: In the present study, Pen ch 13 was further characterized in terms of cDNA cloning, protein purification, enzymatic activity, histamine release and IgE cross reactivity with alkaline serine protease allergens from two other prevalent fungal species--P. citrinum (Pen c 13) and Aspergillus flavus (Asp fl 13). RESULTS: A 1,478-bp cDNA (Pen ch 13) that encodes a 398-amino-acid alkaline serine protease from P. chrysogenum was isolated. This fungal protease has pre- and pro-enzyme sequences. The previously determined N-terminal amino acid sequence of the P. chrysogenum 34-kD major allergen is identical to that of residues 116-125 of the cDNA. Starting from Ala116, the deduced amino acid sequence (283 residues) of the mature alkaline serine protease has a calculated molecular mass of 28.105 kD with two cysteines and two putative N-glycosylation sites. It has 83 and 49% sequence identity with the alkaline serine proteases from P. citrinum and A. fumigatus, respectively. The recombinant Pen ch 13 was recovered from inclusion bodies and isolated under denaturing condition. This recombinant protein reacted with IgE antibodies in serum from an asthmatic patient and with monoclonal antibodies (PCM8, PCM10, PCM39) that reacted with the 34-kD component from P. chrysogenum. The N-terminal amino acid sequence of the purified native Pen ch 13 is identical to that determined previously for the 34 kD major allergen in crude P. chrysogenum extracts. The purified native Pen ch 13 has proteolytic activity with casein as the substrate at pH 8.0. This enzymatic activity was inhibited by phenylmethylsulfonyl fluoride or diethylpyrocarbonate. Pen ch 13 was also able to degrade gelatin and collagen but not elastin. Basophils from 5 asthmatic patients released histamine (12-73%) when exposed to the purified Pen ch 13. In ELISA (enzyme-linked immunosorbent assay) experiments, IgE for Pen ch 13 was able to compete with purified Pen ch 13, Pen c 13 or Asp fl 13 in a dose-related manner. CONCLUSIONS: These results demonstrated that the 34 kD major allergen of P. chrysogenum is an alkaline serine protease. These results also indicated that atopic patients primarily sensitized by either of these prevalent fungal species may develop allergic symptoms by exposure to other environmental fungi due to cross-reacting IgE antibodies against this protease. PMID- 11893852 TI - Identification of cross-reactive proteins amongst different Curvularia species. AB - BACKGROUND: Curvularia lunata is an important inhalant allergen. The present study was undertaken to investigate the shared IgG- and IgE-binding components among seven Curvularia species prevalent in the aerospora. METHODS: Seven different Curvularia species were grown in a semisynthetic medium for 13 days. The extracts were analyzed by SDS-PAGE, immunoblot and ELISA/immunoblot inhibition using sera from C. lunata-positive patients and anti-C. lunata rabbit serum. RESULTS: Different Curvularia species showed 11-19 protein bands on SDS PAGE. Proteins of 12, 20, 31, 45, 53, 78 and 97 kD were present in all the species. Eight out of 98 nasobronchial patients exhibited positive skin tests to C. lunata and to at least five Curvularia species. ELISA using these sera showed IgE binding with Curvularia species. Immunoblot using pooled anti-C. lunata sera from patients showed 5-12 allergenic proteins. Proteins of 12, 31, 45, 53 and 78 kD showed IgE binding in Curvularia species. Antibodies against C. lunata detected 6-14 antigenic proteins on immunoblot. Proteins of 31, 45 and 53 kD showed IgG binding in all the species. Proteins of 31 and 53 kD showed complete IgE/IgG binding inhibition. IgE/IgG ELISA inhibition showed dose-dependent inhibition in Curvularia species. C. lunata extract required 0.17 and 0.11 microg of protein for 50% IgE and IgG inhibition, respectively. C. clavata and C. pallescens required 10 times more protein to exhibit the same inhibition and other species required similar protein levels as those required by C. lunata. CONCLUSIONS: A high degree of cross-reactivity was observed between C. lunata and the six other Curvularia species tested. C. lunata and C. senegalensis shared maximum allergenic and antigenic components. PMID- 11893851 TI - Identification of continuous, allergenic regions of the major shrimp allergen Pen a 1 (tropomyosin). AB - BACKGROUND: Crustaceans and mollusks are a frequent cause of allergic reactions. The only major allergen identified in shrimp is the muscle protein tropomyosin; at least 80% of shrimp-allergic subjects react to tropomyosin. Furthermore, tropomyosin is an important allergen in other crustaceans such as lobsters, crabs and mollusks, as well as other arthropods such as house dust mites and cockroaches, and has been implied as the cause of clinical cross-sensitivity among invertebrates. In contrast, vertebrate tropomyosins are considered non allergenic. OBJECTIVE: The basis of the allergenicity of proteins has not yet been resolved. Thus, tropomyosin molecules provide an excellent opportunity to study the relationship between protein structure and allergenicity. The aim of the current study was to identify the IgE-binding regions of Pen a 1 and compare these regions with homologous sequences in other allergenic and non-allergenic tropomyosins. METHODS: Forty-six overlapping peptides (length: 15 amino acids; offset: 6 amino acids) spanning the entire Pen a 1 molecule were synthesized and tested for IgE antibody reactivity with sera from 18 shrimp-allergic subjects to identify the IgE-binding regions of shrimp tropomyosin. RESULTS: Based on the frequency and intensity of the IgE reactivities, five major IgE-binding regions were identified. All five major IgE-binding regions were 15-38 amino acids long. The major IgE-binding regions identified were: region 1: Pen a 1 (43-57); region 2: Pen a 1 (85-105); region 3: Pen a 1 (133-148); region 4: Pen a 1 (187-202), and region 5: Pen a 1 (247-284). In addition, 22 peptides were categorized as minor IgE-binding regions, and 12 peptides did not bind any IgE antibodies. No substantial differences in amino acid group composition in the five IgE-binding regions compared to the whole molecule were detected. Sequence identities and similarities of the Pen a 1 IgE-binding regions with homologous regions of allergenic arthropod tropomyosins were as high as 100%, whereas identities and similarities with homologous vertebrate sequences ranged from 36 to 76% and 53 to 85%, respectively. CONCLUSION: Five major IgE-binding regions of the allergenic shrimp tropomyosin, Pen a 1, were identified which are positioned at regular intervals of approximately 42 amino acids (7 heptads), suggesting a relationship with the repetitive coiled-coil structure of the tropomyosin molecule. The high degree of similarity between Pen a 1 IgE-binding regions and homologous sequences in invertebrate tropomyosins and the lower percentage of similarity with homologous regions of vertebrate tropomyosins supports a structural basis for cross-reactivity of allergenic tropomyosins. PMID- 11893853 TI - Identification and characterization of Che a 1 allergen from Chenopodium album pollen. AB - BACKGROUND: Pollinosis to Chenopodium album has been reported, but no data are available on its allergenic proteins. METHODS: An allergen from C. album pollen has been isolated by means of gel permeation and reverse-phase high-performance liquid chromatography. Molecular characterization was achieved by concanavalin A reaction, mass spectrometry, Edman degradation and cDNA sequence. Antigenic analyses were performed by immunoblotting, ELISA, and ELISA inhibition, using sera from allergic patients, two Ole e 1-specific monoclonal antibodies and an Ole e 1-specific polyclonal antiserum. RESULTS: The isolated allergen, Che a 1, is a glycoprotein of molecular mass 17.088 kD and 143 amino acid residues, whose sequence exhibits 27-45% identity with known members of the Ole e 1-like protein family. 77% of sera from patients allergic to chenopod pollen were reactive to Che a 1. No correlation was found between the IgE reactivities to Che a 1 and Ole e 1, the major allergens from olive pollen, and both allergens display low, although detectable, IgE and IgG cross-reactivities. CONCLUSIONS: Che a 1, a relevant allergen from chenopod pollen, is structurally related to the Ole e 1 like protein family, but exhibits significant differences on its polypeptide sequence that could explain its different antigenic behavior and limited cross reactivity. PMID- 11893854 TI - Soluble complement receptor type 1 protects rats from lethal shock induced by anti-Crry antibody following lipopolysaccharide priming. AB - BACKGROUND: In rats primed with a trace amount of lipopolysaccharide (LPS), acute lethal shock is induced following the injection of monoclonal antibody against a membrane inhibitor of complement (anti-Crry). Administration of cobra venom factor to exhaust complement before the LPS priming can prevent the lethal reaction. Therefore, we evaluated whether soluble complement receptor type 1 (sCR1), which inhibits complement reaction, can interfere with lethal shock when administered after LPS priming or even after anti-Crry injection. METHODS: sCR1 was administered intravenously before or after the administration of anti-Crry, and the effects on blood pressure and acute lethality were determined. RESULTS: Administration of sCR1 could rescue rats from lethal shock even when it was administered after anti-Crry injection, which immediately causes a blood pressure decrease leading to lethal shock. CONCLUSION: sCR1 may be an effective treatment for acute shock involving complement activation. PMID- 11893855 TI - Trimellitic anhydride-induced cellular infiltration into Guinea pig lung varies with age but not gender. AB - BACKGROUND: In humans the incidence of asthma changes with age and gender. Immature guinea pigs have been used to model the allergic response to the occupational allergen trimellitic anhydride (TMA) where exposure to adults is paramount. We hypothesized that the TMA-induced allergic response in immature guinea pigs was similar to mature animals, regardless of gender. METHODS: Sexually immature and mature female and male guinea pigs were sensitized intradermally with TMA. Three weeks after sensitization they were challenged intratracheally with TMA conjugated to guinea pig serum albumin (TMA-GPSA) or GPSA as a control. Twenty-four hours later cell infiltration into the lung was determined. TMA-specific IgG(1) and IgG(2) were measured in plasma and the complement activation product C3a was measured in the bronchoalveolar lavage fluid. RESULTS: In control animals, numbers of eosinophils and neutrophils varied with age and gender. The TMA-GPSA- induced cellular infiltration was similar in all age/gender groups. However, neutrophils in the lung tissue increased only in immature animals. IgG antibodies differed between groups but did not account for differences in cell infiltration. C3a correlated with the extent of cell infiltration in all groups except mature females. CONCLUSIONS: TMA-induced neutrophilia differs with age. TMA-induced changes in eosinophils and macrophages did not vary with age or gender. The relationship between complement activation and inflammation in mature females differs from that in the other groups, suggesting mediators of the response may change with age and gender. Effects of age and gender need to be considered in animal models of the allergic response. PMID- 11893857 TI - Comparison of IgE antibody concentrations to pyromellitic dianhydride-modified laminin and human serum albumin in sera of exposed workers with respiratory complaints. AB - BACKGROUND: Acid anhydrides are used in a variety of industrial branches because of their highly reactive anhydride groups. Adverse effects of anhydride exposure include toxic/irritative effects as well as IgE- and IgG-mediated respiratory disorders. It was the aim of this study to examine the usefulness of conjugates of pyromellitic dianhydride (PMDA) and laminin, a respiratory tract protein, for the in vitro diagnosis of sensitization to PMDA. METHODS: Sera of PMDA-exposed workers (n = 9) and nonexposed controls (n = 8) were tested with an enzyme allergosorbent test (EAST). Human laminin and human serum albumin (HSA) were derivatized with PMDA and used as solid-phase in a commercial assay. RESULTS: Seven out of 9 exposed workers revealed elevated IgE antibody concentrations (>0.35 kU/l) against laminin-PMDA whereas 5 of 9 subjects had elevated IgE antibody concentrations to HSA-PMDA. All 9 workers had elevated IgE antibody concentrations in at least one of the two tests. Of the 4 workers who complained of shortness of breath, 3 were positive for laminin-PMDA and 2 for HSA-PMDA. All of the nonexposed subjects were negative (<0.35 kU/l) for laminin-PMDA. CONCLUSION: PMDA-modified laminin could provide an additional diagnostic tool for the detection of sensitization to PMDA. PMID- 11893856 TI - Hydrogen peroxide reverses IL-5 afforded eosinophil survival and promotes constitutive human eosinophil apoptosis. AB - BACKGROUND: Eosinophils play a central role in the induction and perpetuation of allergic inflammatory responses. The present study was performed to investigate the effects of reactive oxygen intermediates on constitutive apoptosis as well as on interleukin (IL)-5 afforded human eosinophil survival. METHODS: Peripheral blood eosinophils were isolated by CD16-negative selection to >99% purity and were cultured for 48 h. The number of apoptotic eosinophils in the culture was assessed by flow cytometric analysis of relative DNA content in propidium-iodide stained cells, annexin-V binding or by morphological analysis. Apoptosis was confirmed by the appearance of a typical ladder pattern in the DNA fragmentation assay by agarose gel electrophoresis. RESULTS: Exogenous H(2)O(2) reversed IL-5 afforded eosinophil survival by inducing apoptosis. Constitutive eosinophil apoptosis was inhibited by a reduction of intracellular levels of H(2)O(2) by catalase. Exogenous H(2)O(2) increased the rate of constitutive apoptosis. CONCLUSIONS: Our results suggest that H(2)O(2) may play a role in the downregulation of eosinophilic inflammation by inducing eosinophil apoptosis. PMID- 11893858 TI - Atopic phenotype in children is associated with decreased virus-induced interferon-alpha release. AB - BACKGROUND: Interferon-alpha (IFN-alpha) production in humans is an early event in the nonspecific cellular response to viruses and mediates a wide range of antiviral and immunoregulatory activities. Little is known about the role of IFN alpha in allergic disease. METHODS: In the present study, we performed a retrospective comparative analysis of 88 children with and without an atopic phenotype for virus-induced IFN-alpha production in blood cultures. RESULTS: We were able to demonstrate that patients with allergic asthma (aA) produced significantly lower amounts of virus-induced IFN-alpha than healthy children and patients with nonallergic asthma (naA). Furthermore, the number of eosinophils in atopic children as a marker for allergic inflammation correlated negatively with the IFN-alpha level in blood cultures. Additionally, we found differences between aA and naA patients with respect to the capacity to produce IFN-gamma. Although atopy is thought to be associated with a Th2 cytokine response, in our study, IFN gamma release was not reduced in the allergic children. In contrast, patients with allergic rhinitis showed a significant increase in IFN-gamma release compared to naA patients. CONCLUSIONS: In our study, an early atopic phenotype was related to a reduction in virus induced IFN-alpha release from blood cultures. Thus, after further prospective evaluation, the IFN-alpha level may serve as an additional in vitro marker for the definition of atopy in children. PMID- 11893860 TI - Possible role of nitric oxide and adrenomedullin in bipolar affective disorder. AB - Nitric oxide (NO) has been implicated to play a role in the pathogenesis of depressive disorders. Adrenomedullin (AM) induces vasorelaxation by activating adenylate cyclase and also by stimulating the release of NO. AM immune reactivity is present in the brain, consistent with a role as neurotransmitter. Therefore, it is suggested that these two molecules may play a role together in the brain. We aimed to examine AM and NO in bipolar affective disorder (BPAD). Forty-four patients with BPAD and 21 healthy control subjects were included in this study. DSM-IV diagnosis of bipolar affective disorder (type I, manic episodes) was independently established by two psychiatrists and the Turkish version of the Bech-Rafaelson Mania Scale was administered. Also, a semistructured form was used to ascertain several sociodemographic and clinical variables of the patients. AM and NO were studied in plasma. The mean value of plasma NO levels in the BPAD group of 46.58 +/- 13.97 micromol/l was significantly higher than that of controls (31.81 +/- 8.14 micromol/l) (z = -4.15, p = 0.000). Mean plasma AM levels were found to be increased in patients with BPAD (35.13 +/- 5.26 pmol/l) compared to controls (16.22 +/- 3.02 pmol/l) (z = -6.16, p = 0.000). AM levels of BPAD patients were approximately 2-fold higher than controls. AM levels were positively correlated with the duration of hospitalization for the current episode and negatively correlated with the total duration of illness. Both NO and AM may have a pathophysiological role in BPAD (type I, manic episodes) and the clinical symptomatology and prognosis of BPAD. PMID- 11893859 TI - Intravenous ulinastatin therapy for Stevens-Johnson syndrome and toxic epidermal necrolysis in pediatric patients. Three case reports. AB - BACKGROUND: More effective therapy is needed for the treatment of Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). The clinical efficacy of intravenous ulinastatin therapy was investigated in 3 Japanese pediatric patients with SJS or TEN. METHODS: Ulinastatin was given to 1 pediatric SJS patient and 2 pediatric TEN patients within 7 days (patient 1; SJS), 6 days (patient 2; TEN), or 4 days (patient 3; TEN) after the onset of the skin rash. Ulinastatin was administered intravenously at a dose of 7,500 U/kg/day (maximum dose: 300,000 U/day). No corticosteroids were given. After the skin lesions resolved, the ulinastatin dose was reduced to between 2,500 and 5,000 U/kg/day as maintenance therapy and then the drug was withdrawn. RESULTS: Erythema, fatigue, and fever improved within 12-36 h of starting the ulinastatin infusion, and the skin lesions resolved completely after 4-7 days of ulinastatin therapy. None of the patients had cutaneous or ocular sequelae. No patient developed secondary infection or relapse and ulinastatin therapy caused no side effects. CONCLUSION: Ulinastatin dramatically reduced the febrile period with no adverse effects and was very safe in this study. Ulinastatin appears to be a useful and effective therapy for controlling SJS and TEN without sequelae. PMID- 11893861 TI - Change in the equilibrium of the levels of free and bound forms of alpha1 microglobulin and ulinastatin in patients with mood disorders. AB - We have previously found that the relationship between the urinary content of alpha1-microglobulin (alpha1M) and ulinastatin (UT) in patients with mood disorders (MD) differs from that in age-matched healthy subjects. Similar results were obtained for the relationship between the content of free forms of alpha1M and UT in serum, and that between the ratio of the content of free form to the total (free + bound forms) content (F/T ratio) of alpha1M and UT in serum. As for the content of alpha1M and UT in serum, statistical differences were not observed between MD patients and healthy subjects with regard to the respective total content of alpha1M and UT nor the F/T ratio of alpha1M, whereas the F/T ratio of UT was significantly (p < 0.05) lower in MD patients than in healthy subjects. Furthermore, the serum cortisol content was higher in MD patients than in healthy subjects, and a positive correlation between cortisol content and F/T ratios of UT was demonstrated in the present investigation. These results suggest that the equilibrium of free and bound forms of UT in serum, but not of alpha1M, tends to decrease the free-form level of UT in MD patients. The efficiency of cortisol, which might increase the free-form level of UT, markedly deteriorated in MD patients. PMID- 11893862 TI - Opioid addiction changes cerebral blood flow symmetry. AB - Changes in regional cerebral blood flow (rCBF) due to long-term abuse of opioids such as heroin or morphine are not yet fully understood in humans. The goal of the present study was to investigate rCBF alterations in a large sample of long term opioid addicts in comparison to healthy controls. We investigated 21 opioid dependent subjects, who were currently abusing heroin or were enrolled in a methadone or morphine maintenance program, and 36 healthy controls with (99m)Tc HMPAO single photon emission computed tomography. We found a decrease in rCBF in most regions of interest in patients in comparison to controls. Long-term opioid dependence seems to decrease prefrontal CBF in particular. A right-greater-than left CBF asymmetry in healthy subjects was reversed in patients. This change in CBF symmetry could reflect the different emotional status of opioid-dependent patients. Our findings are in line with neuropsychological investigations indicating a correlation of mood states with lateralization of hemispheric activation patterns. PMID- 11893863 TI - Short-term cognitive improvement in schizophrenics treated with typical and atypical neuroleptics. AB - OBJECTIVE: Atypical neuroleptics seem to be more beneficial than typical ones with respect to long-term neuropsychological functioning. Thus, most studies focus on the long-term effects of neuroleptics. We were interested in whether atypical neuroleptic treatment is also superior to typical drugs over relatively short periods of time. METHODS: We studied 20 schizophrenic patients [10 males, mean age 35.5 years, mean Brief Psychiatric Rating Scale (BPRS) score at entry 58.9] admitted to our hospital with acute psychotic exacerbation. Nine of them were treated with typical and 11 with atypical neuroleptics. In addition, 14 healthy drug-free subjects (6 males, mean age 31.2 years) were enrolled in the study and compared to the patients. As neuropsychological tools, a divided attention test, the Vienna reaction time test, the Benton visual retention test, digit span and a Multiple Choice Word Fluency Test (MWT-B) were used during the first week after admission, within the third week and before discharge (approximately 3 months). RESULTS: Patients scored significantly worse than healthy controls on nearly all tests (except Vienna reaction time). Clinical ratings [BPRS and Positive and Negative Symptom Scale for Schizophrenia (PANSS)] improved markedly (p < 0.01), without a significant difference between typical and atypical medication. Clinical improvement (PANSS total score) correlated with less mistakes on the Benton test (r = 0.762, p = 0.017) and an improvement on the divided attention task (r = 0.705, p = 0.034). Neuropsychological functioning (explicit memory, p < 0.01; divided attention, p < 0.05) moderately improved for both groups under treatment but without a significant difference between atypical and typical antipsychotic drugs. CONCLUSIONS: Over short periods of time (3 months), neuropsychological disturbances in schizophrenia seem to be moderately responsive to both typical and atypical neuroleptics. PMID- 11893864 TI - Plasma catecholamines and selective slow wave sleep deprivation. AB - The present study evaluated the effect of slow wave sleep (SWS) deprivation on plasma levels of catecholamines in healthy male volunteers. Eleven volunteers spent 4 nights in the sleep laboratory (2 nights of habituation and 2 further nights); during the latter, 1 night served as control, and in the other, SWS deprivation was performed. Blood was drawn at 30-min intervals. SWS was reduced by 86%; no sleep stage 4 was observed during the SWS-deprived nights. SWS reduction was found not to correlate with catecholamine levels. However, epinephrine levels were found to be sensitive to sleep fragmentation. The time interval between arousals in the SWS-deprived night as well as the difference in sleep efficiency were related to increases in epinephrine levels (p < 0.01 and p < 0.025, respectively). These results support the view that continuity rather than the duration of SWS is important for the recuperative value of sleep. PMID- 11893866 TI - The cyclic alternating pattern decreases as a consequence of total sleep deprivation and correlates with EEG arousals. AB - The aim of this study was to assess the sensitivity of the cyclic alternating pattern (CAP) parameters to the increased need for recuperation that follows one night of sleep deprivation. Nine normal male subjects were recorded for 3 nights [adaptation, baseline (BSL) and recovery (REC)]. BSL and REC nights were separated by a 40-hour sleep deprivation. Recovery after sleep deprivation was characterized by a decrease in stage 1, stage 2 and sleep latency, and an increase in slow wave sleep (SWS) and the sleep efficiency index, while rapid eye movement (REM) and non-REM (NREM) time did not differ as compared to BSL sleep. CAP parameters were significantly affected by sleep deprivation, and the main results were the following: (1) the CAP rate (time spent in CAP phases/NREM time) significantly decreased from 38.85 to 31.47%, and (2) within phase A subtypes, there was a significant decrease in A3 subtypes (from 38.11 to 19.57). Furthermore, the number of arousals scored according to American Sleep Disorders Association rules strongly correlated with A3 subtypes during both the BSL (r = 0.79) and REC nights (r = 0.95). These results indicate that recuperative processes after sleep deprivation are also associated with a lesser arousal instability as defined by the reduction of the CAP rate, which is strongly correlated with EEG arousals. PMID- 11893865 TI - Examination of GABAergic and dopaminergic compounds in the acquisition of nicotine-conditioned hyperactivity in rats. AB - In rats, a distinct environment repeatedly paired with nicotine (0.421 mg/kg base, s.c.) comes to evoke an increase in activity in the absence of any drug. This hyperactivity indicates a Pavlovian-conditioned association between the environment and nicotine. We investigated whether a dopamine D(1) receptor antagonist (SCH-23390), a D(2)/D(3) antagonist (eticlopride) or a GABA(B) agonist (baclofen) would prevent the acquisition of nicotine-conditioned hyperactivity. In saline-pretreated rats, acute nicotine suppressed activity during the conditioning phase (i.e. environment-nicotine pairings); chronic nicotine stimulated activity. Pretreatment with SCH-23390 (0.01 mg/kg, i.p.) attenuated the activating effects of nicotine without affecting controls. Eticlopride (0.03 0.07 mg/kg, i.p.) and baclofen (0.625 and 1.25 mg/kg, i.p.) did not affect nicotine-induced activity in a selective manner. Regardless of the pretreatment drug, rats acquired the environment-nicotine association as indexed in a drug free test. The inability of SCH-23390 to block the acquisition of nicotine conditioned locomotor activity is notable because in past research SCH-23390 blocked expression of the learned association. PMID- 11893867 TI - A study on gender and age differences in sleep spindles. AB - In the present work, gender differences in sleep spindle topography were examined in 40 subjects. Their median age was 32 years (range 22-49 years). Spindles were detected from 3,306,060 s of visually scored stage 2 sleep EEG by a previously validated automatic fuzzy detector at 1-second intervals. A total of 271,168 spindles were found from the six EEG channels analyzed. Females showed a significantly higher percentage of spindles in the left frontal channel than males (Fp1-A2; p = 0.026). To confirm that this difference was gender and not age related, the subjects were divided into two age groups. No significant differences in spindle activity of the frontal channels were found between the groups. However, the interindividual spindle variability seemed to be at least as large as that stemming from gender. PMID- 11893868 TI - Changes in brain complexity during valproate treatment in patients with partial epilepsy. AB - OBJECTIVE: The effect of valproate (VPA) on human electroencephalography (EEG) was studied using nonlinear dynamics analysis to investigate changes in brain complexity. METHODS: We propose a spatial linear mode complexity (SLMC) measure to quantify the complexity of spatial linear modes in multichannel EEGs. Nine patients with complex partial seizures who had not previously been exposed to antiepileptic drugs (AEDs) were included in this study. Eighteen-channel EEG data were collected before and after VPA therapy. Changes in brain complexity were examined using the proposed SLMC measure, which reflects brain complexity. Fifteen normal, healthy subjects were included as a control group. To compare SLMC with spectral analysis, we performed spectral analysis within the conventional frequency bands. RESULTS: Spectral analysis showed that the patient group had decreased relative power of the alpha2 band in the T7, P3, O1 and C4 leads before VPA treatment and an increased relative theta power in the O1 lead relative to the control group. However, no significant changes occurred in any lead at any frequency band after VPA treatment. The mean SLMC value was significantly lower in the patient group before treatment than in the control group (p = 0.026). The average SLMC value for all patients increased after treatment and neared that of the control group, although statistical significance was not attained (p = 0.074). CONCLUSIONS: These results suggest that epilepsy patients have interictal abnormalities that are demonstrated by reduced brain complexity, and that VPA partially reverses this trend. Nonlinear analysis of EEG data may be useful in evaluating the effect of AEDs. PMID- 11893870 TI - Substance P and affective disorders: new treatment opportunities by neurokinin 1 receptor antagonists? AB - Substance P (SP) is a neuropeptide which is abundant in the periphery and the central nervous system, where it is colocalized with other neurotransmitters such as serotonin or dopamine. SP has been proposed to play a role in the regulation of pain including migraine and fibromyalgia, asthma, inflammatory bowel disease, emesis, psoriasis as well as in central nervous system disorders. This review summarizes our current knowledge of the role of SP in the pathogenesis of neuropsychiatric disorders with special emphasis on affective disorders including bipolar disorders. It also reviews current treatment approaches with neurokinin 1 receptor antagonists which appear to be promising drugs for the future treatment of affective disorders. PMID- 11893871 TI - Sleep and sleep-wake manipulations in bipolar depression. AB - In the last 30 years, it has been convincingly demonstrated that sleep in major depression is characterized by disturbances of sleep continuity, a reduction of slow wave sleep, a disinhibition of REM sleep including a shortening of REM latency (i.e. the time between sleep onset and the occurrence of the first REM period) and an increase in REM density. Furthermore, manipulations of the sleep wake cycle like total or partial sleep deprivation or phase advance of the sleep period have been proven to be effective therapeutic strategies for patients with unipolar depression. The database concerning sleep and sleep-wake manipulations in bipolar disorder in comparison is not yet as extensive. Studies investigating sleep in bipolar depression suggest that during the depressed phase sleep shows the same stigmata as in unipolar depression. During the hypomanic or manic phase, sleep is even more curtailed, though subjectively not experienced as disturbing by the patients. REM sleep disinhibition is present as well. An important issue is the question, whether sleep-wake manipulations can also be applied in patients with bipolar depression. Work by others and our own studies indicate that sleep deprivation and a phase advance of the sleep period can be used to treat bipolar patients during the depressed phase. The risk of a switch into hypomania or mania does not seem to be more pronounced than the risk with typical pharmacological antidepressant treatment. For patients with mania, sleep deprivation is not an adequate treatment--in contrast, treatment strategies aiming at stabilizing a regular sleep-wake schedule are indicated. PMID- 11893872 TI - Neurobiological findings before and during successful lithium therapy of a patient with 48-hour rapid-cycling bipolar disorder. AB - 48-hour rapid cycling is a very rare form of bipolar disorder, characterized by regular periodic changes of mood from one day to the other. We report on a patient who suffered from a 48-hour rapid cycling without a history of bipolar disorder before the abrupt onset of his rapid mood cycles. We present polysomnographic and neuroendocrine findings and the clinical course based on daily self-ratings of mood. Treatment with lithium carbonate effectively reduced the amplitude of the mood cycles. With plasma levels between 0.8 and 1.0 mmol, almost complete remission occurred. An overview on previous reports on the therapeutic effect of mood stabilizers in this rare form of bipolar disorder is presented. PMID- 11893873 TI - Clinical relevance and treatment possibilities of bipolar rapid cycling. AB - Bipolar rapid cycling (RC) is defined as 4 or more affective episodes within 1 year. It has been postulated that RC is related to a poor response to lithium, to the same extent as mixed episodes or other atypical symptoms of the illness. This article reviews the current status of alternative pharmacological or otherwise supportive therapies of RC. Biological parameters and characteristics of the illness associated with RC like gender prevalence in women, hyperthyroidism, catecholamine-O-methyltransferase allele, the influence of sleep, different subtypes of bipolar disorder and the risk of antidepressant-induced cycling will be discussed in detail. PMID- 11893874 TI - Comparison of long-term monitoring methods for bipolar affective disorder. AB - Long-term monitoring is a clinical necessity in bipolar affective disorder. The most important requirements are usability and value for clinical decisions. Four methods of long-term monitoring - (1) the Adjective Mood Scale by von Zerssen, (2) Kraepelin's early life charts, (3) the NIMH Life Chart Method by Leverich and Post and (4) the Social Rhythm Metric by Monk - were compared regarding the acceptance by patients, the time commitment needed for documentation and training as well as the usability for the clinician regarding psychotherapy and decision support in pharmacotherapy. The Adjective Mood Scale is easiest to learn, Kraepelin's life chart offers the best graphical presentation, the NIMH life charts offer the best clinical decision support and the Social Rhythm Metric integrates a psychotherapeutic approach. PMID- 11893875 TI - Bupropion as add-on strategy in difficult-to-treat bipolar depressive patients. AB - Bupropion, a selective norepinephrine and dopamine reuptake inhibitor, has been suggested for the treatment of bipolar depression, not only because of its efficacy, but also because of a probably lower risk of inducing switches to hypomania or mania. Most studies on bupropion treatment in bipolar patients have been performed in moderately ill out-patients. In contrast, we report on a sample of difficult-to-treat, predominantly severely ill, co-morbid, psychotic or therapy-refractory bipolar depressive in-patients. In this open and prospective study, 13 patients were treated with bupropion as an add-on strategy mainly to other antidepressants and to various mood stabilizers. Our data support the idea that bupropion is a first-line antidepressant in the treatment of severe bipolar depression. Eight of 13 patients showed a >50% reduction of Montgomery-Asberg Depression Scale ratings within 4 weeks. Co-medication with drugs commonly used in treatment-resistant bipolar disorder including venlafaxine, clozapine, lithium, topiramate and sodium valproate was safe in our small sample. While adhering to the suggestion of Goren and Levin not to exceed a daily dose of 450 mg of bupropion when treating bipolar depressed patients, we did not observe any switch from depression to hypomania or mania. PMID- 11893876 TI - Clozapine as add-on medication in the maintenance treatment of bipolar and schizoaffective disorders. A case series. AB - Atypical neuroleptics are increasingly used in the treatment of bipolar and schizoaffective disorders. Currently, numerous controlled short-term studies are available for clozapine, olanzapine, risperidone or quetiapine, but long-term data are still missing. Three patients (2 with bipolar disorder, 1 with schizoaffective disorder) are described who showed a marked reduction of affective symptomatology after clozapine had been added to mood stabilizer pretreatment. The patients were seen once a month before and after the introduction of clozapine for at least 6 months. Treatment response was evaluated using different rating scales (IDS, YMRS; GAF; CGI-BP) and the NIMH Life Chart Methodology. All patients showed a marked improvement after the add-on treatment with clozapine had been initiated. Clozapine was tolerated well with only transient and moderate weight gain and fatigue as only side effects. This case series underlines the safety and efficacy of clozapine as add-on medication in the treatment of bipolar and schizoaffective disorders. PMID- 11893877 TI - Carcinogen-induced site-specific mutagenesis and genetic susceptibility in squamous cell carcinoma of the head and neck. AB - Tobacco and alcohol have been identified as the most important risk factors for squamous cell carcinomas of the head and neck (HNSCC). Especially for tobacco, some of the carcinogen-induced mutations that trigger transformation have been identified. Much attempt has been made to show a relationship between the mutations of growth regulatory genes, i.e. tumor suppressor- or oncogenes, and the exposure to specific exogeneous mutagens. One of the best examined genes - harboring the most DNA damages (DNA adducts) - caused by xenobiotics is p53. This tumor suppressor gene is an important regulator of cell cycle and apoptosis and is mutated in 40-60% of all HNSCC. Further studies link specific mutations of the ras oncogene to definite carcinogens. The question of why these mutations lead to cancer in some smokers but not in others, i.e. the individual's susceptibility to external carcinogens, remains unclear. One possibility is the individual's ability (or non-ability) of detoxifying carcinogenic xenobiotics or repairing the DNA damages they caused. There are several known polymorphisms of enzymes involved in detoxification as well as in DNA repair - which might explain the interindividual variable susceptibility to HNSCC in smokers and drinkers. However, the results of several examined polymorphisms are diverse or even controversial. The diversity might be due to ethnical differences, study populations, tumor localizations as well as intratumoral heterogeneity. In this review, we attempt to discuss the carcinogen-specific mutations as well as the genetic polymorphisms, which may transfer an enhanced susceptibility to suffer HNSCC. PMID- 11893878 TI - Management of germ cell tumors in children: approaches to cure. AB - The introduction of cisplatinum chemotherapy and current advances in the surgical treatment have resulted in a dramatic improvement of the prognosis of children with malignant germ cell tumors (GCT). Cisplatinum chemotherapy generally results in sufficient systemic tumor control, but local relapses may still occur in patients who did not receive adequate local treatment. Therefore, the therapeutic consideration must take into account age, primary site of the tumor, and its histology. In gonadal tumors, there is a high chance of primary complete resection since these tumors tend to be encapsulated, and particularly testicular GCT are often detected at a low tumor stage. In contrast, a primary complete resection may be impossible in large nongonadal tumors such as sacrococcygeal or mediastinal GCT. In these tumors, a neoadjuvant or preoperative chemotherapy after clinical diagnosis by imaging and evaluation of tumor markers significantly facilitates complete resection on delayed surgery. In addition, the impact of chemotherapy on local tumor control may be enhanced by locoregional hyperthermia. In most intracranial GCT complete resection is impossible and may be associated with significant morbidity. Nevertheless, biopsy is essential for diagnosis in nonsecreting tumors. In intracranial GCT, radiotherapy significantly contributes to local tumor control, and doses are stratified according to histology. These general considerations have been integrated into national and international cooperative treatment protocols. In most current protocols, treatment is stratified according to an initial risk assessment that includes the parameters age, site, histology, stage, completeness of resection and the tumor markers alpha(1)-fetoprotein (AFP) and human choriogonadotropin (beta-HCG). With such modern protocols overall cure rates above 80% can be achieved. Moreover, the previously high-risk groups may now expect a favorable prognosis with this risk adapted treatment, whereas an increasing number of low-risk patients are treated expectantly or with significantly reduced chemotherapy. As current biologic studies reveal distinct genetic patterns in childhood GCT, it can be expected that further combined clinical and genetic studies will be valuable for risk assessment of childhood GCT. PMID- 11893879 TI - Hereditary premenopausal breast cancer. AB - Less than 1% of breast cancers occur between the age of 20 and 30 years, but more than 50% of breast cancers under the age of 30 years are hereditary. Breast cancer mainly occurs sporadically, however, in 5 to maximally 10% of cases a genetic predisposition is present. Mutations in the already sequenced tumor suppressor genes BRCA1 and BRCA2 account for 60-70% of these hereditary breast cancers. The chromosomal location of BRCA1 is 17q21 and that of BRCA2 is 13q12 13. Screening procedures and possible prevention strategies for women with mutations in the BRCA1 and BRCA2 genes are discussed. They include the use of tamoxifen. PMID- 11893880 TI - Functional magnetic resonance imaging - basics and applications in oncology. AB - Functional MR imaging (F-MRI) permits to visualize active processes evolving in the human brain. The shifts in blood oxygenation following local neuronal activation are used for generating an MR signal. Specific procedural problems have to be solved when using F-MRI in patients with brain neoplasms. The relationship between perirolandic tumors and the adjacent motor areas has been studied most intensively. When the resection of tumors in the motor area is considered, functional imaging can help the surgeon to avoid or at least to minimize postoperative functional deficits. Overall, F-MRI study results are useful in at least one crucial preoperative decision step for about 90% of brain tumor patients. Experience with functional imaging in radiotherapy of brain neoplasms is still limited. Presently, F-MRI cannot yet replace intraoperative brain mapping. PMID- 11893881 TI - Use of hematopoietic growth factors in elderly patients receiving cytotoxic chemotherapy. AB - Myelosuppression is a common side effect in elderly patients undergoing chemotherapy. Neutropenia and anemia cause considerable morbidity, may increase mortality, and can result in a worse outcome of treatment in elderly patients compared to younger patients with comparable type and stage of disease. The availability and proven efficacy of hematopoietic growth factors such as granulocyte colony-stimulating factor (G-CSF) or granulocyte-macrophage colony stimulating factor (GM-CSF) and erythropoietin (EPO) have had a considerable impact on supportive care in cancer patients: Several randomized trials have demonstrated a reduction of neutropenia and the frequency of severe infections in elderly patients treated with G-CSF following myelotoxic chemotherapy compared with patients without growth factor support. Both for G-CSF and for recombinant human erythropoietin (rHu-EPO) several studies have demonstrated the safety and effectiveness of these molecules in elderly patients with regard to increasing hemoglobin concentrations, improving quality of life (rHu-EPO), and neutrophil recovery. Although a positive effect of the use of growth factors on overall survival in elderly cancer patients is not yet proven, a reduction of chemotherapy-induced side effects could clearly be shown. The National Comprehensive Cancer Network (NCCN) of cancer centers has recommended that all patients aged 70 years and older treated with CHOP or cytotoxic chemotherapy of comparable intensity should receive prophylactic G-CSF administration, and that the hemoglobin concentration be maintained at >or=12 g/dl in elderly patients undergoing chemotherapy. PMID- 11893882 TI - Analysis of minimal sample volumes from head and neck cancer by laser scanning cytometry. AB - BACKGROUND: The increasing diversity in therapeutic strategies in head and neck oncology is dependent on the development of equally appropriate diagnostic tools. A growing number of diagnostic procedures is intended to be performed on an out patient basis. In this context, analyses of hypocellular specimens such as fine needle aspirate biopsies (FNABs) or swabs are very important: There are minimal side-effects, and they can be analysed within hours. MATERIAL AND METHODS: Laser scanning microscopy (LSC) is a microscope-based method combining the advantages of flow cytometry and image analysis: In addition to the fluorescence data of each individual cell, its morphology can be documented by re-staining with a conventional cytological staining. Any cell can then be re-localised in the microscope for direct observation. FNABs and swabs are incubated in PBS, erythrocytes are lysed, and cells are mounted on slides. After fixation in ethanol, cells are stained for cytokeratin by indirect immunolabelling and for DNA by propidium iodide. Analysis by LSC is performed to determine the ploidy of the epithelial cells. For immunophenotyping of peripheral blood in cancer patients by LSC 20 microl full blood are stained for CD antigens by direct immunolabelling and for DNA by 7-aminoactinomycin D. RESULTS: FNABs and swabs were taken from 150 malignancies of different sites in total; all specimens yielded sufficient cells (>5,000). 30 tumours of the parotid gland were analysed in detail: Out of 9 malignant tumours 8 showed aneuploidy, whereas all 21 benign tumours were diploid. Immunophenotyping in 23 tumour patients showed a significant reduction of lymphocytes in the peripheral blood as compared to healthy individuals. CONCLUSIONS: Further studies have to be performed to validate the analysis of hypocellular specimens by LSC and to determine its role in routine clinical work. Its potential is most evident in tumours that are not accessible for open biopsy such as those of the parotid gland or the larynx. PMID- 11893883 TI - Gemcitabine monotherapy as second-line treatment in cisplatin-refractory transitional cell carcinoma - prognostic factors for response and improvement of quality of life. AB - OBJECTIVES: i) To evaluate objective response, toxicity, and quality of life (QoL) of gemcitabine monotherapy as second-line treatment in patients with cisplatin-refractory, metastatic transitional cell carcinoma (TCC). ii) To assess prognostic parameters for response to treatment and for improvement of QoL parameters. PATIENTS AND METHODS: 30 patients were prospectively enrolled in this open-label, nonrandomized multicenter phase II trial. Patients received up to 6 courses of gemcitabine monotherapy (1,250 mg/m(2) on day 1 and 8 of a 21-day course). 28 of 30 patients were available for response evaluation. RESULTS: Objective response (OR) was seen in 3/28 (11%) of patients (2 complete remissions, 1 partial remission). The mean time to progression (TTP) was 4.9 +/- 3.5 months and mean disease-specific survival time was 8.7 +/- 4.7 months. 13 of 28 patients did not progress (OR + 10 stable diseases), and TTP (8.0 +/- 2.7 months, p < 0.001) as well as survival time (10.2 +/- 3.8 months, p < 0.05) differed significantly from those who showed progressive disease within 18 weeks of treatment. Pain values significantly improved in the group of responders from 4.3 +/- 1.9 to 5.8 +/- 1.3 points (p < 0.05). Response to cisplatin pretreatment was the best prognosticator for the response to gemcitabine. CONCLUSIONS: Gemcitabine monotherapy as second-line treatment is justified in patients with metastatic TCC who are refractory to cisplatin treatment. Patients with initially OR to cisplatin benefit most from second-line treatment. QoL remains stable during treatment, and pain improves especially in patients with bone metastases. PMID- 11893884 TI - Radiochemotherapy for anal carcinoma - effectivity and late toxicity. AB - BACKGROUND: Radiochemotherapy of anal carcinoma is an organ-sparing approach with a high curative potential. The purpose was to evaluate the effectivity and late toxicity for patients treated with radiochemotherapy in our department. PATIENTS AND METHODS: During 1990-2000, 27 patients with anal carcinoma were treated at the Universitatsklinikum Mannheim. The median follow-up time was 23 months (max. 68 months). Before treatment, 23 patients were colostomy-free. Patients were treated according to 3 different protocols (Cummings n = 8, EORTC n = 5, RTOG n = 14). Acute toxicity was scored according to the RTOG/EORTC scale, and late toxicity according to the LENT/SOMA scale. RESULTS: 25 patients completed the therapy. One patient died due to leukopenic sepsis, and 1 patient interrupted therapy. 4 patients had a tumor relapse (2 patients immunosuppressed, 1 T4 tumor and 1 recurrence at the field-margin), one underwent abdomino- perineal resection. This resulted in a disease-free survival of about 80% and colostomy free survival of 90% at 5 years. Total doses < 50 Gy and immunosuppression resulted in higher recurrence rates. Most patients suffered from acute toxicity grades 2 (n = 7) and 3 (n = 19) and late toxicity grades 1 (n = 7) and 2 (n = 4). CONCLUSIONS: Radiochemotherapy for anal carcinoma is an effective therapy with acceptable toxicity. Immunosuppressed patients have a higher risk for tumor recurrences and should be monitored more closely. PMID- 11893885 TI - Weekly irinotecan in a patient with metastatic colorectal cancer on hemodialysis due to chronic renal failure. AB - BACKGROUND: The cytotoxic treatment of patients suffering from advanced or metastatic cancer undergoing hemodialysis due to chronic renal failure still remains a problem, since for those patients pharmacokinetic and pharmacodynamic data on most cytotoxic agents are lacking. CASE REPORT: We report a 45-year-old male who suffered from chronic renal failure and was diagnosed with stage-3 colorectal cancer (CRC) in February 2000. After surgical removal of the tumor an adjuvant chemotherapy of dose-reduced i.v. bolus 5-fluorouracil and folinic acid was begun (Mayo protocol). Due to excessive gastrointestinal toxicity, therapy was discontinued after the first cycle. In April 2000 liver metastases were diagnosed. The patient was then put on a weekly schedule of dose-reduced CPT-11 (50 mg/m(2), 80 mg total). No hematological or non-hematological toxicity grade 3/4 was observed. Due to excellent tolerability and lack of severe side effects the dose was increased up to 80 mg/m(2) (140 mg total) weekly. A dose escalation to 100 mg/m(2) (180 mg total) resulted in severe diarrhea (grade 4). Within 2 months of treatment the patient achieved a lasting partial remission until April 2001 (12 months). A significant progression of hepatic metastases required an alternative treatment regimen beginning in July 2001 (HAI, hepatic artery infusion). CONCLUSION: This case report demonstrates the feasibility and efficacy of a weekly treatment with dose-reduced CPT-11 in a patient with metastatic CRC on hemodialysis due to chronic renal failure. PMID- 11893886 TI - Intramedullary cavernous angiomas of the spinal cord in the pediatric age group: a pediatric series. AB - The authors have reviewed available data from 7 pediatric patients with intramedullary spinal cord cavernous angioma (ISCCA) reported in the literature, and added from their own series 2 pediatric patients, for a total of 9 patients. This group of pediatric patients' clinical presentation, course, management and outcome were compared to their adult counterparts as reported in the literature. In contrast to adults, children with symptomatic ISCCA do not show a gender imbalance and the thoracic spinal cord is not predominantly involved. Pediatric patients commonly present with an acute episode and rapid deterioration. A more favorable outcome has been reported in children as compared to adults in the face of relatively similar presenting deficits. As in adults, magnetic resonance imaging (MRI) remains the diagnostic and postoperative test of choice. Complete resection affords the best chance for cure. Symptomatic children with ISCCA characteristically present with an acute deficit and rapid deterioration. MRI of the entire neuraxis is recommended for lesion multiplicity. An attempt at total resection and long-term MRI follow-up are recommended. PMID- 11893887 TI - Predictors of outcome following traumatic brain injury in young children. AB - The relationship between clinical and neuroimaging variables and multiple outcome measures was examined in a longitudinal, prospective study of 60 children less than 6 years of age who sustained either inflicted or noninflicted traumatic brain injury. Hierarchical multiple regression indicated that the modified Glasgow Coma Scale score, the duration of impaired consciousness and the number of intracranial lesions visualized on CT/MRI accounted for a significant amount of the variance in the Glasgow Outcome Scale (GOS), cognitive and motor scores at baseline, 3- and 12-month evaluations. Inflicted brain injury adversely affected both GOS and cognitive outcomes. Pupillary abnormalities were associated with poorer motor outcome. Neither age at injury nor the Injury Severity Score accounted for significant variability in outcomes. PMID- 11893888 TI - Cerebral salt wasting syndrome following brain injury in three pediatric patients: suggestions for rapid diagnosis and therapy. AB - The association between hyponatremia and intracranial pathology has been well described. When accompanied by natriuresis, hyponatremia has most commonly been attributed to inappropriate secretion of antidiuretic hormone. However, there is growing evidence to suggest that many of these patients may actually have cerebral mediated salt losses, a disorder referred to as the cerebral salt wasting syndrome (CSWS). While this syndrome has been reasonably well described in adults, data regarding CSWS in pediatric-aged patients remains sparse. Since fluid management of these disorders is different, it is important that the clinician be able to rapidly differentiate between them. We report three cases of CSWS in acutely brain-injured children and comment on the role that early quantitation of urine volume and urine sodium concentration had in rapidly establishing the correct diagnosis. PMID- 11893889 TI - Pediatric craniopharyngiomas: clinicomorphological study of 189 cases. AB - OBJECTIVE: To understand the pathological changes in pediatric craniopharyngiomas in order to determine the diagnosis and operative strategy and to prevent damage to the hypothalamus. METHODS: A total of 189 cases of pediatric craniopharyngiomas from 1990 to 1998 were reviewed and analyzed based on computerized tomography scans, magnetic resonance imaging and operations. RESULTS: Of the 189 cases, 187 (98.9%) were cystic tumors and calcification could be seen in 176 cases (93.1%). Two cases were solid tumors (1.1%), and calcification occurred in only 1 of these. There was a gliosis layer between the wall of the tumor and the hypothalamus. CONCLUSION: Cystic changes and calcification are the pathological features of pediatric craniopharyngiomas. There are some special relationships between the tumors and stalk. This is the basis for the total removal of pediatric craniopharyngiomas. PMID- 11893890 TI - Robot-assisted endoscopic intrauterine myelomeningocele repair: a feasibility study. AB - BACKGROUND: Animal experiments have suggested that the intrauterine environment causes secondary injury to the congenitally dysplastic spinal cord. This in turn suggests that early closure of the myelomeningocele sac might prevent secondary injury and therefore improve neurologic outcome. This study was designed to examine the technical feasibility of performing intrauterine myelomeningocele repair using a robot-assisted endoscopic system in an animal model. METHODS: Six fetal sheep underwent creation and repair of a full-thickness skin lesion using the da Vinci system. RESULTS: With the device's advanced articulated instruments and three-dimensional optics, it was possible to endoscopically repair the induced skin defects. CONCLUSION: We conclude that, with the recent evolution in robotics and minimally invasive techniques, intrauterine endoscopic surgery has become a realistic goal that promises to reduce the associated risks of fetal surgery and extend the indications for its use. PMID- 11893891 TI - Cervical hematomyelia: a rare entity in a neonate with cesarean section and surgical recovery. AB - Spinal cord injury with or without trauma has been reported in the perinatal period. The prognosis depends primarily on diagnosis of the level, extent and nature of the lesion, established by correlations between clinical, imaging and electrophysiological data. A 25-day-old boy with normal birth weight delivered at term by cesarean section was transferred to Inonu University Turgut Ozal Medical Center because of respiratory distress and brachial diplegia. A suspicious medullary lesion on cervical computerized tomography was confirmed as an intramedullary lesion extending from C3 to D1 on magnetic resonance imaging (MRI). Emergent surgery consisting of exposure of the lesion site and interlaminar direct puncture of the lesion under fluoroscopy revealed that the pathology was an intramedullary hematoma. The partial evacuation of the lesion with direct puncture, the patient's neurological improvement and close follow-up of the patient with ultrasonography, electrophysiology and MRI are discussed in the light of recent literature. PMID- 11893892 TI - Growing fractures of the orbital roof. A report of two cases and a review. AB - Growing fractures rarely arise in the skull base. Only six cases of orbital roof growing fractures were found in the relevant literature. We report two such cases. The first case was a 2-year-old girl who had progressive proptosis for 6 months following a mild head injury 1 year previously. The second case was a 9 year-old girl with a history of injury at the age of 3 months. She developed eye deviation and proptosis for 1 year. Computed tomography scan is excellent for demonstrating bony defects in the orbital roof, while magnetic resonance imaging is more sensitive in showing the intraorbital extension of a leptomeningeal cyst. Both patients were operated successfully and proptosis disappeared postoperatively. The exact pathophysiology of growing fractures is still debated in the literature, but a dural laceration along a fracture line is noted in all cases, and frontobasal brain injury seems to play an important role in the pathogenesis of the fracture growth. Growing fractures of the orbital roof should be suspected if ocular symptoms appear in a child who had sustained a head injury several months or years before. PMID- 11893893 TI - Acute hydrocephalus following a Chiari I decompression. AB - Complications of posterior cranial fossa decompression for Chiari malformations are uncommon. We present three patients who developed infratentorial supracerebellar hygromas causing acute hydrocephalus after posterior cranial fossa decompression. PMID- 11893894 TI - Cervical intramedullary gliofibroma in a child: a case report and review of the literature. AB - A rare case of intramedullary gliofibroma in a 12-year-old girl is reported. MRI revealed an intramedullary tumor with two different intensities, and these portions showed different staining patterns after gadolinium contrast enhancement, with faint enhancement at the periphery and strong enhancement in the central portion. Intraoperatively, the tumor had a relatively clear margin to the spinal cord and was composed of two portions of different consistencies, one soft in the periphery and the other elastic and firm in the central portion. This tumor was grossly, totally removed. The patient made a full recovery and there was no recurrence 2 years and 9 months after operation. There have only been 7 previous case reports of spinal intramedullary gliofibroma. Gliofibroma is usually a relatively benign astroglial variant. Recognition of such a variant is clinically important in considering the extent of the operation and postoperative adjuvant therapy. PMID- 11893895 TI - Gliomatosis cerebri. PMID- 11893896 TI - Regulation of endothelin-1 production in deoxycorticosterone acetate- salt treated endothelial cells. AB - The aim of this study was to investigate the direct effect of deoxycorticosterone acetate (DOCA) on the expression of endothelin-1 (ET-1) mRNA and production of ET 1 peptides in cultured bovine carotid endothelial cells (BCEC) and human umbilical vein endothelial cells (HUVEC). The mineralocorticoid, DOCA, was administered to BCEC and HUVEC with or without additional salt (200 mmol/l). The ET-1 mRNA was elevated to 150% and 180% after a 24-hour incubation with DOCA (5 micromol/l) in BCEC and HUVEC, respectively. Intracellular content of ET-1 peptides of BCEC and HUVEC was increased from 21.66 +/- 0.3 to 23.64 +/- 0.19 fmol/10(5) cells and from 8.38 +/- 0.82 to 11.26 +/- 0.91 fmol/10(5) cells, respectively, in the DOCA-conditioned medium. Also, DOCA treatment for 24 h increased the secretion of ET-1 peptide from 1.62 +/- 0.17 to 5.33 +/- 0.67 fmol/10(5) cells in BCEC and from 0.95 +/- 0.08 to 3.56 +/- 0.36 fmol/10(5) cells in HUVEC. In DOCA-salt-treated endothelium, regulation of ET-1 gene was similar to that with DOCA alone. The DOCA-induced increase in expression of ET-1 mRNA and the ET-1 peptide level were both diminished in dexamethasone (DEX, 10 nmol/l)- or mifepristone (RU486, 0.5 micromol/l)-treated endothelium. These results suggest that DOCA directly increased the ET-1 mRNA expression in endothelium and could be mediated in part by a glucocorticoid receptor pathway which was independent of salt and hyperosmolarity. PMID- 11893897 TI - Effects of SM-20550, a new Na(+)/H(+) exchange inhibitor, on survival and infarct size in rat myocardial infarction. AB - The effect of a novel potent Na(+)/H(+) exchange inhibitor, SM-20550 [N (aminoiminomethyl)-1,4-dimethyl-1H-indole-2-carboxamide methanesulfonic acid], on survival after myocardial infarction was studied. Anesthetized rats underwent occlusion of the coronary artery (30 min) followed by reperfusion (14 days). SM 20550 was administered intravenously before ischemia (1-day treatment group) or before ischemia and on the 2 days following (3-day treatment group). The infarct size was significantly reduced on the 14th day after surgery by approximately 17 and 20% in 1- and 3-day treatment groups, respectively. The survival rate on day 14 was significantly enhanced in both treatment groups (96%) compared with the vehicle-treated control group (70%). These results suggest that SM-20550 improved survival after myocardial infarction, at least due to its antinecrotic effect. PMID- 11893898 TI - Acetylcholine-induced contractions in the perforating branch of the human internal mammary artery: protective role of the vascular endothelium. AB - The effect of acetylcholine (ACh) on the isolated, nonprecontracted perforating branch of the human internal mammary artery (HIMA) was investigated. ACh induced concentration-dependent contractions of nonprecontracted rings with denuded endothelium (pEC(50) = 6.72 +/- 0.02, E(max) = 88.8% of contractions induced by phenylephrine, 10(-5) mol/l) and was without effect on arterial segments with intact endothelium. An inhibitor of nitric oxide synthase, N(G)-monomethyl-L arginine (L-NMMA), or indometacin, a cyclooxygenase inhibitor, had no effect on acetylcholine-induced contractions of rings of the perforating branch of HIMA with denuded endothelium (pEC(50) = 6.76 +/- 0.03 and 6.62 +/- 0.05, respectively). In the presence of indometacin, ACh did not evoke contractions of arterial segments with intact endothelium. In contrast, in the same type of preparations ACh induced contractions in the presence of L-NMMA (E(max) = 34%). The muscarinic receptor antagonists atropine (no selectivity), pirenzepine (M(1)), methoctramine (M(2)), and p-fluoro-hexahydro-sila-difenidol (M(1)/M(3)) competitively antagonized the response to ACh. The pA(2) values were 9.60 +/- 0.10, 6.99 +/- 0.02, 6.37 +/- 0.17, and 8.02 +/- 0.06, respectively. In conclusion, the results obtained indicate that secretion of nitric oxide from vascular endothelium may protect the perforating branch of HIMA against the contractile effects of ACh. On the basis of differential antagonist affinity, it can be suggested that the muscarinic receptors involved in the ACh-induced contractions of the isolated perforating branch of the HIMA are predominantly of the M(3) subtype. PMID- 11893899 TI - Effects of Y-27632, a Rho/Rho kinase inhibitor, on leukotriene D(4)- and histamine-induced airflow obstruction and airway microvascular leakage in guinea pigs in vivo. AB - Recent in vitro studies have shown that the Rho/Rho kinase pathway is involved in the mechanism of not only airway smooth muscle contraction but also vascular endothelial permeability caused by certain stimuli. This suggests that Rho/Rho kinase inhibitors may become useful agents against asthma via reduction of increased airway microvascular leakage, one of the main features of this disease. Thus, we wanted to know the in vivo effect of Y-27632, a selective Rho kinase inhibitor, on airway microvascular leakage caused by leukotriene D(4) (LTD(4)) and histamine, potent mediators of allergic airway inflammation, by comparing its effect against airflow obstruction. For comparison, the effects of procaterol, a beta(2)-adrenoceptor agonist, on these responses were also studied. Tracheostomized guinea pigs were given either aerosolized Y-27632 (3 or 15 mmol/l), procaterol (6 micromol/l) or vehicle (0.9% NaCl) for 5 min under spontaneous breathing. After being mechanically ventilated, the animals were given intravenous Evans blue dye 15 min after the end of inhalation. One minute later, either 2 nmol/kg LTD(4), 300 nmol/kg histamine or vehicle was administered intravenously. After measurements of lung resistance (R(L)) for 6 min, the lungs of animals were taken out, and the amount of extravasated Evans blue dye was examined as an index of leakage. Inhaled Y-27632 dose-dependently attenuated increases in R(L) caused by LTD(4) and histamine. The degree of inhibition was almost similar between 15 mmol/l Y-27632 and 6 micromol/l procaterol. By contrast, only 15 mmol/l, but not 3 mmol/l, Y-27632 partially reduced LTD(4) induced leakage. Histamine-induced Evans blue dye extravasation was not inhibited by 15 mmol/l Y-27632. Procaterol significantly inhibited the dye extravasation caused by either LTD(4) or histamine. These results suggest that Y-27632 is not a useful agent in attenuating airway microvascular leakage which is seen in asthma, although it is potent in inhibiting airflow obstruction. PMID- 11893900 TI - Effect of captopril on urinary kallikrein, blood pressure and myocardial hypertrophy in diabetic spontaneously hypertensive rats. AB - We investigated the total urinary kallikrein levels, left-ventricular wall thickness and mean arterial blood pressure of nontreated and captopril-treated diabetic and nondiabetic spontaneously hypertensive rats. The mean arterial blood pressure was significantly elevated in diabetic spontaneously hypertensive rats as compared to nondiabetic spontaneously hypertensive rats. Captopril treatment caused a significant reduction in the arterial blood pressure of both nondiabetic and diabetic spontaneously hypertensive rats. The left-ventricular wall thickness was also significantly reduced in diabetic and nondiabetic spontaneously hypertensive treated with captopril as compared to nontreated diabetic and nondiabetic spontaneously hypertensive rats. The total urinary kallikrein levels were significantly raised in captopril-treated diabetic and nondiabetic spontaneously hypertensive rats against the values obtained from nontreated diabetic and nondiabetic spontaneously hypertensive rats. These results indicate that blood pressure reduction and left ventricular wall regression with captopril treatment might be due to enhanced renal kallikrein formation. The significance of these findings is discussed. PMID- 11893902 TI - Influence of estrogen and/or progesterone on isolated ovariectomized rat uterus. Responsiveness to Ang II. AB - Steroids hormones can influence several functions of the uterus, including agonist-induced contractions. The aim of this work is to study the influence of hormonal replacement on angiotensin II (Ang II) and losartan responsiveness on the isolated rat uterus. The Ang II pD(2) values are: 9.69 +/- 0.07 for vehicle treated animals, 8.85 +/- 0.06 for estrogen-treated animals, 10.12 +/- 0.03 for progesterone-treated animals and 8.90 +/- 0.03 for estrogen-and-progesterone treated animals. The losartan pD(2)' values are: 8.43 +/- 0.03 for vehicle treated animals, 8.21 +/- 0.03 for estrogen-treated animals, 7.83 +/- 0.05 for progesterone-treated animals and 8.70 +/- 0.09 for estrogen-and-progesterone treated animals. There is not a correlation between Ang II pD(2) and losartan pD(2)' values, suggesting that the hormones affect Ang II and losartan binding by different mechanisms in rat uterus. PMID- 11893901 TI - Effect of alpha-adrenoceptor agonists and antagonists on imipramine-induced antinociception in the rat formalin test. AB - In this study, the effect of central administration of the alpha-adrenoceptor agents on the antinociception induced by imipramine in the formalin test has been investigated. Intraperitoneal (IP) administration of different doses of imipramine (10-80 mg/kg) and intracerebroventricular (ICV) administration of the alpha(2)-adrenoceptor agonist clonidine (0.05-0.8 microg/rat) elicited a dose dependent antinociception in the both phases of the test. Furthermore, different doses of clonidine (0.05-0.2 microg/rat) increased the antinociception induced by imipramine (10 and 20 mg/kg). The alpha(2)-adrenoceptor antagonist yohimbine (2 microg/rat, ICV) reduced the response of a low dose imipramine (10 mg/kg, IP) plus different doses of clonidine (0.05, 0.1 and 0.2 microg/rat, ICV), but did not alter the response induced by higher doses of imipramine (20 and 40 mg/kg) alone or in combination with clonidine. Yohimbine by itself elicited no effect. The alpha(1)-adrenoceptor agonist phenylephrine (0.07-1.5 microg/rat) induced antinociception in both phases of the formalin test, but did not alter the imipramine-induced antinociception. The alpha(1)-adrenoceptor antagonist prazosin neither elicited antinociception nor altered the imipramine response. Yohimbine (2 microg/ rat, ICV) in combination with prazosin (0.5 microg/rat, ICV) caused more inhibition of the response of imipramine or imipramine plus clonidine. Therefore, it is concluded that alpha(2)-adrenoceptor mechanism may be involved in the imipramine-induced antinociception. PMID- 11893903 TI - Role of G(s) proteins in hypoxic constriction of sheep pulmonary artery rings. AB - The introduction of hypoxia is well known to cause contraction of pulmonary artery rings in vitro. Despite intensive studies, the cellular mechanisms of hypoxic pulmonary vasoconstriction are still not well defined. In this study, we aimed to determine the contribution of G(S) proteins in hypoxia-induced vasoconstriction in large-diameter sheep pulmonary arteries using cholera toxin (CT). Hypoxia caused further contractions in serotonin but not in NaF precontracted pulmonary artery rings. However, hypoxic vasoconstriction due to lowering of pO(2) from 97 to 5 mm Hg was totally abolished by preincubation with CT in serotonin-precontracted arteries. These preliminary results indicate that signal transduction mediated by G(s) proteins may be an important mechanism in the hypoxic vasoconstriction of isolated pulmonary arteries of sheep. PMID- 11893904 TI - The ISOBM TD-7 Workshop on hCG and related molecules. Towards user-oriented standardization of pregnancy and tumor diagnosis: assignment of epitopes to the three-dimensional structure of diagnostically and commercially relevant monoclonal antibodies directed against human chorionic gonadotropin and derivatives. AB - The ISOBM TD-7 hCG Workshop was established to characterize the molecular epitope structure and specificities of a panel of diagnostically relevant monoclonal antibodies (MAbs) directed against human chorionic gonadotropin (hCG) and its derivatives, and to consider how this information could be used to improve comparability of immunoassay results for these analytes. In this multicenter study, 27 MAbs have been characterized in detail as to their main and fine specificities by direct binding-, competitive- and sandwich-RIA, -ELISA, BIAcore and Western blotting. Antigens used in the study included the upcoming first WHO reference reagents for immunoassay, i.e. nick-free hCG (hCG), nicked hCG (hCGn), hCG alpha-subunit (hCGalpha), hCG beta-subunit (hCGbeta), nicked hCG beta-subunit (hCGbetan), hCG beta-core fragment (hCGbetacf), synthetic peptides of hCGbeta C terminal peptide (hCGbetaCTP), and homologous hormones, luteinizing hormone (LH) and subunits (LHbeta) from various species. Correct classification of blinded internal controls demonstrated the reliability of the MAb referencing approach. Three-dimensional molecular epitope assignment was possible in many instances by comparing immunoreactivity of the ISOBM MAbs (n = 27) to a large panel of MAbs (n = 18) previously well characterized in the Innsbruck (P.B.) and Paris (J.M.B.) laboratories. All three major antibody specificities (alpha, n = 1; beta, n = 21; alphabeta, n = 5) were represented in the TD-7 MAb panel. HCGbeta MAbs could further be subdivided into (i) those recognizing hCGbeta only (epitopes: beta(6), n = 1; beta(7), n = 2; beta(14), n = 1) and (ii) those recognizing hCGbeta + hCG (beta1, beta2, beta4, beta5, n = 10; beta8 and beta9, n = 9). Members of the latter group were specific either for hCG + hCGbeta + hCGbetacf (beta1, n = 3) or hCG + hCGbeta + hCGbetaCTP (beta8, n = 6; beta9, n = 1) or in addition to hCG + hCGbeta + hCGbetacf recognized hLH/hLHbeta to a minor (beta2, n = 3; beta4, n = 3) or similar degree (beta5, n = 1). Epitopes were (i) located on the first and third loops protruding from the cystine knot of hCGbeta (beta2-beta6, aa hCGbeta20-25 and 68-77), (ii) presumably centered around the knot itself (beta1), or (iii) on hCGbetaCTP (epitope beta8 = hCGbeta141-144, beta9 = hCGbeta113-116). The ISOBM panel of MAbs represents all major epitope specificities suitable for the design of specific sandwich immunoassays. High analyte variability in serum and urine during the course of pregnancy and tumor development favors certain epitope combinations. For routine diagnostic purposes, assays recognizing a broad spectrum of hCG/hCGbeta variants such as hCG + hCGn + hCGbeta + hCGbetan + hCGbetacf + -CTPhCG + -CTPhCGbeta may be useful. Low cross-reactivity against related glycoprotein hormones (e.g. hLH) and their derivatives is mandatory. These criteria are best met by combinations of MAbs directed against epitopes located around the cystine knot (beta1) and against those encompassing the top of loops 1 and 3 on hCGbeta (beta2, beta4). The first WHO reference reagents for immunoassay of hCG and hCG-related molecules being prepared by the IFCC should facilitate characterization of what assays for 'hCG' are measuring. The next step towards improving between-laboratory comparability of measurements of hCG/hCG derivatives in pregnancy and oncology is provided by results of this TD-7 Workshop. PMID- 11893905 TI - S-100 protein serum levels in patients with benign and malignant diseases: false positive results related to liver and renal function. AB - S-100 serum concentrations were analyzed in 39 healthy people, 130 patients with benign diseases and 304 patients with malignancies, including 49 patients with locoregional diseases and 255 with advanced diseases. S-100 was determined by a commercial immunoluminometric assay, and 0.20 ng/ml was considered to be the upper limit of normality. In none of the healthy people was S-100 higher than 0.2 ng/ml. Slightly high S-100 concentrations were found in 33 out of 130 patients (25%) with benign diseases (mean 0.21 +/- 0.45 ng/ml). Significantly higher S-100 serum levels were found in patients with liver cirrhosis (63%, 10/16) (p = 0.024) or renal failure (45%, 8/18) (p = 0.03) than in patients with other benign diseases or in healthy people. Abnormal S-100 serum levels were found in 68 of the patients (22.5%) with malignancy (mean 1.01 +/- 5.9 ng/ml). The highest S-100 concentrations were found in patients with malignant melanomas (p = 0.001). Excluding melanoma patients, the S-100 serum levels in patients with malignancies were not related to tumor origin or stage but were clearly related to the site of metastasis, with patients with liver metastases showing higher values than patients with metastases without liver involvement (p = 0.02). No statistical differences were found among patients with liver cirrhosis, primary liver cancer or liver metastases. In conclusion, S-100 is a useful marker for melanoma, but abnormal levels of this tumor marker may be found in benign and malignant diseases associated with liver or renal injury. PMID- 11893906 TI - Frequent alterations of the beta-catenin protein in cancer of the uterine cervix. AB - Cancer of the uterine cervix is still the leading cause of death among women with cancer in developing countries. Although infections with human papillomavirus are necessary, other molecular alterations that are needed at the cellular level for development of these tumors remain largely unknown. Beta-catenin is a key regulator located within the Wnt signaling cascade whose alterations constitute an important event in colon carcinogenesis. In many malignancies increased levels of the beta-catenin protein have been found, associated with its nuclear and/or cytoplasmic accumulation. To search for possible alterations of this pathway we examined the expression and localization of the beta-catenin protein in tumors from the uterine cervix and cell lines derived from them. Beta-catenin was found accumulated in the cytoplasm and/or nuclei of 12 out of 32 samples. In accordance, increased levels of this protein were observed in 9 out of 20 tumors analyzed. Importantly, PCR-SSCP and sequence analysis showed no mutations in exons 3, 4 and 6 of the beta-catenin gene. Our findings indicate that alterations of beta-catenin are frequent in these tumors and suggest that they may play an important role in the development of cancer of the uterine cervix. They also indicate that higher protein levels and abnormal localization may result from several different mechanisms. PMID- 11893907 TI - De novo expression of the Muc2 gene in pancreas carcinoma cells is triggered by promoter demethylation. AB - It has been established that mucin-producing variants of different subtypes of pancreatic carcinomas, including the intraductal papillary and ductal mucinous tumors, have usually a more favorable prognosis. Intraductal papillary and ductal mucinous tumors have also been shown to ectopically express the intestinal mucin gene MUC2. The mechanism of the de novo expression of this gene in tumors may have potential implications for the modulation of its behavior. We studied, therefore, the mechanism of the de novo expression of MUC2 in pancreas carcinoma cells in vitro. The MUC2 gene promoter is methylated in the nonexpressing pancreatic cell line PANC-1 and is not methylated in the expressing cell line BxPC-3. The promoter is silenced by methylation as shown by reporter expression assays. De novo expression of MUC2 in PANC-1 cells is triggered by treating the cells with a pharmacological inhibitor of DNA methylation (5-aza-2' deoxycytidine). There was no decrease or loss of expression of the methyltransferase DNMT1 in the MUC2-producing cells. These data show that the de novo expression of the MUC2 gene in pancreas carcinoma cells is associated with promoter demethylation. They warrant further investigations on the relationship between MUC2 promoter demethylation in pancreatic cancer and the prognosis of carcinoma patients. PMID- 11893908 TI - Increased sensitivity to nicotine-induced seizures in mice heterozygous for the L250T mutation in the alpha7 nicotinic acetylcholine receptor. AB - alpha7 Nicotinic acetylcholine receptors (nAChRs) are sparsely distributed throughout the peripheral and central nervous systems. Several studies have suggested that central alpha7 nicotinic receptors may influence sensitivity to nicotine-induced seizures in mice. In order to investigate the effect of alpha7 nAChRs on seizure sensitivity, we tested heterozygous mice with a threonine for leucine substitution at position 250 (L250T) within the channel domain, which is known to increase current amplitude and decreases desensitization of the channel. We show that administration of low doses of nicotine to these mutant mice increased the sensitivity to nicotine-induced seizures and the mortality rate. EEG recordings showed high amplitude rhythmic activity during tonic-clonic seizures. Pretreatment with the alpha7 nicotinic receptor antagonist methyllycaconitine inhibited the seizures induced by nicotine. These findings further suggest an important role for alpha7 nAChRs in the nicotine-induced seizures model of epilepsy. PMID- 11893909 TI - Attenuation of SAH-induced cerebral vasospasm by a selective ECE inhibitor. AB - CGS 26303, a dual inhibitor of endothelin-converting enzyme-1 (ECE-1) and neutral endopeptidase 24.11, was previously shown to prevent and reverse vasospasm in an experimental model of subarachnoid hemorrhage (SAH). However, reversal of the vasospastic response was not very efficacious. This study was designed to examine the effects of a highly selective ECE-1 inhibitor, CGS 35066, on SAH-induced cerbrovasospasm. Experimental SAH was induced in New Zealand white rabbits by injecting autogenous blood into cisterna magna and CGS 35066 was injected i.v. twice daily, either at 1 h (prevention protocol) or 24 h (reversal protocol) after SAH. Treatment with CGS 35066 significantly attenuated basilar arterial narrowing at a dose of 1 mg/kg in both protocols. These findings provide support for the use of selective ECE-1 inhibitors for the treatment of SAH-induced vasospasm even after the process of arterial narrowing has begun. PMID- 11893910 TI - Birth insult and stress interact to alter dopamine transporter binding in rat brain. AB - This study investigated whether mild birth complications (C-section birth, C section + 15 min global anoxia) interact with stress at adulthood to modulate levels of [3H]WIN 35428 binding to dopamine transporters (DAT) in rat brain. Without stress, adult C-sectioned rats showed increased DAT binding in the dorsal striatum and nucleus accumbens core compared to vaginal birth, while anoxic rats showed increased DAT binding in cingulate and infralimbic cortices. Stress at adulthood had differential effects on DAT binding in the three birth groups. Thus, after repeated tail pinch stress at adulthood, DAT binding was significantly lower in the nucleus accumbens in both the C-section group and the anoxic group, compared to vaginal birth. It is concluded that a history of birth complications can alter the manner in which DAT is regulated by stress in the adult rat brain. PMID- 11893911 TI - Remodeling of somotasensory hand representations following cerebral lesions in humans. AB - There is evidence of reorganization of somatotopic maps following cortical lesions in mammals such as monkeys, raccoons and rats. However, there has been a striking lack of research on somatosensory plasticity following cerebral damage in adult humans. We describe two individuals with left hemisphere damage who misperceive the locations of tactile stimuli whose presence or absence they can readily detect. We find that the mislocalizations preserve the relative topography of pre-lesion experiences, resulting in shifted and compressed representations of the hand surfaces. These results not only provide evidence for systematic remodeling of somatotopic maps in humans, they also reveal that the systematic changes in cortical topography that have been documented using electrophysiological methods may give rise to similarly systematic changes in somatosensory perception itself. PMID- 11893912 TI - Therapeutic window for nicotinamide following transient focal cerebral ischemia. AB - The therapeutic window with the neuroprotectant nicotinamide (NAm) was tested in a model of stroke. Either 2, 4 or 6 h after the onset of transient (2 h) focal cerebral ischemia, Wistar rats received either saline or NAm (500 mg/kg). Sensory and motor behavioral scores and weight of the animals were obtained before surgery, and 2 h, 3 and 7 days after stroke onset. Cerebral infarct volumes were measured on day 7 after sacrifice. NAm given 4 or 6 h after stroke onset significantly (p<0.05) reduced the cerebral infarction and improved the behavioral scores, respectively, compared to saline-injected animals. There was a non-significant improvement in weight gained by NAm-treated rats at 3 and 7 days following stroke compared to the saline-injected controls. PMID- 11893913 TI - Aspirin inhibits stress-induced increase in plasma glutamate, brain oxidative damage and ATP fall in rats. AB - The precise mechanisms by which stress induces brain damage are still being elucidated. The high-output, inducible isoform of nitric oxide (NO) synthase (iNOS) is expressed in rat brain after immobilisation stress and its inhibition protects against cell damage in this condition. We have hereby explored some mechanisms involved in iNOS expression and studied the effects of aspirin, a NSAID with neuroprotective actions, in this model. Acute (6 h) stress exposure in rats caused brain expression of iNOS, an increase in plasma glutamate and brain TNF-alpha, induction of oxidative indicators in brain and a fall in brain ATP levels. Prior administration of aspirin (10 mg/kg i.p.) inhibited all these effects caused by stress, suggesting possible therapeutic implications of this drug in this condition. PMID- 11893915 TI - Electrical stimulation enhances the survival of axotomized retinal ganglion cells in vivo. AB - In view of recent reports on the survival promotion of damaged spiral ganglion cells and motoneurons by electrical stimulation, we hypothesized that an electrical stimulation of the cut optic nerve (ON) may promote the survival of axotomized retinal ganglion cells (RGCs) in vivo. To test this hypothesis, we examined 1 week after ON transection the RGC densities in the retinas with or without electrical stimulation. The densities of surviving RGCs in the retinas with the electrical stimulation increased as compared with those without the electrical stimulation. We concluded that electrical stimulation of the ON enhances the survival of axotomized RGCs in vivo, probably due to electrical activation of their soma. PMID- 11893916 TI - Rapid mapping of finger representations in human primary somatosensory cortex applying neuromagnetic steady-state responses. AB - We analyzed somatosensory evoked steady-state fields in order to localize finger representations in the hand area of the primary somatosensory cortex (S1). Using a 122-channel whole-head neuromagnetometer we recorded in six healthy subjects neuromagnetic responses to high frequency electrical stimuli delivered simultaneously to digit I, II, III and V at 22, 24, 27 and 30 Hz, respectively, and to transient stimulation of each single digit with a frequency of 3 Hz. Responses were averaged separately for each digit and were modeled by single equivalent current dipoles. Both conditions yielded the typical somatotopic finger representations within S1 hand area. Dipole locations did not differ significantly between the transient and the steady-state stimulation. Therefore, simultaneous high-frequency stimulation of the digits seems to be a reliable method for rapid and detailed mapping of the S1 hand area. This procedure has potential advantages over recording of transient responses. With simultaneous steady-state stimulation the measurement times are reduced to 2 min for mapping the whole hand area. Because of this our method probably increases spatial accuracy and permits repeated short interval recordings, e.g. in experiments studying short term plasticity. PMID- 11893914 TI - State-dependent cross-inhibition between anionic GABA(A) and glycine ionotropic receptors in rat hippocampal CA1 neurons. AB - Using whole-cell patch-clamp recording of acutely isolated rat hippocampal CA1 neurons, we studied the state-dependent cross-inhibition between anionic GABA(A) and glycine (Gly) ionotropic receptors. Co-application of relatively high concentrations of GABA and Gly produced a total current (IGABA + Gly) much smaller than the linear summation of the two individual responses. The mutual inhibition between IGABA and IGly was voltage-independent. However, no current inhibition was observed with co-application of low concentrations of these two agonists. The hippocampal neurons lacking Gly response (IGly) showed no current inhibition with GABA current (IGABA), whereas the neurons losing GABA response (IGABA) (after complete rundown of IGABA) showed no current inhibition with IGly. The current inhibition was also absent in the presence of Gly or GABA(A) receptor antagonists, strychnine or bicuculline, respectively. The results indicate that cross-inhibition between GABA(A) and Gly receptors is state-dependent and a direct receptor-receptor interaction might enable the cross-inhibition to occur. PMID- 11893917 TI - Basic fibroblast growth factor induces TGF-beta release in an isoform and glioma specific manner. AB - The aim of the current study was to determine whether basic fibroblast growth factor (bFGF), regulates the release of transforming growth factor-beta1 (TGF beta1) from C6 glioma cells. The results of the study show that bFGF (2, 5 and 10 ng/ml) dose dependently induced the release of TGF-beta1 from C6 glioma cells, with the 10 ng/ml dose inducing a 2- to 3-fold increase of TGF-beta1 levels. This effect was evident as early as 6 h following treatment, with maximal levels observed at 18 h. The effect of bFGF was largely on latent TGF-beta1, and was isoform specific, as bFGF had no effect on TGF-beta2 release. The bFGF effect on TGF-beta1 was also glioma specific, as no such stimulatory effect was observed in rat cortical astrocytes. PMID- 11893918 TI - Induction of Cdk5 activity in rat skeletal muscle after nerve injury. AB - Cyclin-dependent kinase 5 (Cdk5) was originally identified as a serine/threonine kinase and subsequently demonstrated to play a critical role in the development of CNS. We recently reported the novel function of Cdk5 in the neuregulin signaling pathway during the development of neuromuscular junction (NMJ). Here, we report the regulation of Cdk5 and p35 in rat skeletal muscle after nerve injury. Northern blot analysis revealed that Cdk5 and p35 transcripts were up regulated in muscle after nerve denervation. The temporal profiles for the regulation of Cdk5 and p35 transcripts were different, suggesting that these changes in gene transcription might be regulated by different mechanism. Our finding on the ability of tetrodotoxin to induce p35 transcript in muscle suggested that electrical activity could regulate p35 expression. In addition to the induction of mRNA expression, the total Cdk5 and p35-associated kinase activity in muscle increased prominently after nerve denervation. Taken together, our findings suggest that Cdk5 and p35 may play important physiological roles in muscle regeneration following nerve injury. PMID- 11893920 TI - Role of human frontal and supplementary eye fields in double step saccades. AB - Transcranial magnetic stimulation (TMS) was applied over the frontal eye field (FEF) and the supplementary eye field (SEF) regions before execution of the first saccade in a double step task. When applied over the FEF, stimulation increased the percentage error in amplitude of the contralateral second saccade as compared to no stimulation. This was due to an interference with retinotopic but not craniotopic gain calculation. Stimulation of the SEF region interfered with saccade ordering. Thus, FEF might participate in target memorization whereas SEF is confirmed to code order information for sequential saccades even in this paradigm of only two consecutive movements. PMID- 11893919 TI - Feeling touches in someone else's hand. AB - Cerebral damage may induce a delusional belief so that patients claim that their limbs contralateral to the side of the lesion belong to someone else (somatoparaphrenia). This disorder, which is not due to a general delirium, is frequently accompanied by the inability to feel tactile sensations in the 'non belonging' part of the body. We report the unique case of a patient with somatoparaphrenia in whom dense tactile imperception in the left hand dramatically recovered when she was instructed to report touches delivered to her niece's hand, rather than to her own hand. We suggest that, through this verbal instruction, the mismatch between the patient's belief about the ownership of her left hand and her ability to perceive touch on it was transiently recomposed. This is evidence that apparently elementary deficits, such as hemianesthesia, and selective delusional behavior, such as somatoparaphrenia, may both originate from an impairment of the body image. PMID- 11893921 TI - Distribution of fibroblast growth factor receptors and their co-localization with vasopressin in the choroid plexus epithelium. AB - Increasing evidence indicates that basic fibroblast growth factor (FGF2), a well known mitogen, can regulate the synthesis and secretion of peptide hormones. FGF2 has also been recently shown to inhibit cerebrospinal fluid (CSF) formation and increase the number of dark choroid plexus epithelial cells. These latter FGF2 actions could be mediated by vasopressin (VP) synthesized by and released from choroidal epithelium. The present study was therefore designed to determine whether VP co-localizes with fibroblast growth factor receptors (FGFRs) in the choroid plexus epithelium. Using confocal laser-scanning microscopy, we demonstrated the apical (CSF-facing) distribution of FGFRs in epithelial cells. FGFRs were expressed on clusters of VP-positive cells, with the intensity of FGFR immunopositive staining varying from one group of cells to the other. These observations are in line with the possible mediatory role of VP in the FGF2 dependent inhibition of CSF secretion and the induction of dark epithelial cells. PMID- 11893924 TI - A LIM-homeodomain code for development and evolution of forebrain connectivity. PMID- 11893922 TI - Role of Ca(v) 2.3 (alpha1E ) Ca2+ channel in ischemic neuronal injury. AB - We investigated the role of the Ca(v)2.3 (alpha1E) channel in ischemic neuronal injury using Ca(v)2.3 mutant mice. In focal ischemia model with a complete occlusion of the middle cerebral artery in vivo, infarct at 24 h was significantly larger in Ca(v)2.3 mutant mice compared with that in wild-type controls. In vitro Ca2+ imaging studies using hippocampal slices revealed that oxygen-glucose deprivation induced a [Ca2+]i increase in the hippocampal CA1 region more vigorously in Ca(v)2.3 mutant mice than in wild-type controls, and that tetrodotoxin or bicuculline application abolished the difference between the genotypes. These results suggest that the Ca(v)2.3 channel plays a protective role in ischemic neuronal injury by a mechanism in which GABAergic neuronal actions are involved. PMID- 11893925 TI - Addictive drugs modify excitatory synaptic control of midbrain dopamine cells. PMID- 11893927 TI - The APACHE II score is unreliable to diagnose necrotizing pancreatitis on admission to hospital. AB - INTRODUCTION: The APACHE II score is highly recommended worldwide for the assessment of severe pancreatitis (interstitial and necrotizing), and a score of at least eight points on admission to the hospital is said to indicate severe pancreatitis. AIM: To evaluate this assumption and to check whether an APACHE II score of at least eight points really indicates necrotizing pancreatitis as shown by contrast-enhanced computed tomography (CT). METHODOLOGY: This study included 326 patients with a first attack of acute pancreatitis and is part of a prospective study on the natural course of acute pancreatitis. All patients underwent contrast-enhanced CT within 72 hours of admission. The following parameters for the severity of the disease were used: respiratory and renal failure according to the Atlanta classification; indication for dialysis, ventilation, and surgery; time spent in intensive care unit and total hospital stay; Ranson score adjusted for cause; Imrie score; and Balthazar score (CT). RESULTS: Of the 326 patients, 262 (80%) had interstitial pancreatitis and 64 (20%) had necrotizing pancreatitis. In 74 (28%) of the 262 patients with interstitial pancreatitis, the APACHE II score was at least eight points, indicating severe pancreatitis (overestimation of the disease), whereas the score was less than eight in 41 (64%) of 64 patients with necrotizing pancreatitis (underestimation). Sensitivity was 36%; specificity was 72%; the positive predictive value was 24%; and the negative predictive value was 82%. CONCLUSION: The evaluation of sensitivity, specificity, and positive and negative predictive value for all APACHE II score points showed that there was not a "golden" cutoff to detect necrotizing pancreatitis. We conclude that the APACHE II score on admission to the hospital is unreliable to diagnose necrotizing pancreatitis. PMID- 11893928 TI - Acute pancreatitis in five European countries: etiology and mortality. AB - INTRODUCTION: In recent years, many advances have been made in the diagnosis and treatment of acute pancreatitis that have lead to a significant reduction in both morbidity and mortality; however, knowledge of the etiology and of the relation between etiology and mortality is far from complete. AIM: To obtain a more comprehensive view of the etiology and mortality of acute pancreatitis in Europe than has been given by previous single-center studies. METHODOLOGY: The study comprised 1,068 patients in five European countries who were admitted to hospitals for acute pancreatitis from January 1990 to December 1994. Data for each patient were collected on a standardized form. RESULTS: Of the 1,068 patients (692 men, 376 women; mean age, 52.8 years; range, 10-95 years), 589 had edematous pancreatitis, and 479 the necrotic form. Cholelithiasis (37.1%) and alcohol (41.0%) were the most frequent etiologic factors. In Germany, cholelithiasis and alcohol occurred with similar frequency (34.9 and 37.9%, respectively); in Hungary, alcohol predominates over cholelithiasis (60.7 vs. 24.0%); in France, a small predominance of alcohol was seen (38.5 vs. 24.6%); and in Greece and Italy, there was a clear predominance of cholelithiasis over alcohol (71.4 vs. 6.0% and 60.3 vs. 13.2%, respectively). The differences in the frequency of cholelithiasis and alcohol between Greece and Italy and the other countries were statistically significant (p < 0.01). Eighty-three patients (7.8%) died of acute pancreatitis; 77 (16.1%) had necrotic disease and 6 (1.0%) edematous. There was no statistically significant difference in mortality among the etiologic groups, and no relation was found between mortality and age. CONCLUSION: Both cholelithiasis and alcohol were main etiologic factors in the more northern countries studied, whereas cholelithiasis alone predominated in the more southern ones. Mortality was high for necrotic pancreatitis; it was similar among the various etiologic groups, and there was no relationship between mortality and age. PMID- 11893929 TI - Glucosamine-induced beta-cell dysfunction: a possible involvement of glucokinase or glucose-transporter type 2. AB - INTRODUCTION: The mechanism for beta-cell dysfunction induced by glucosamine has not yet fully been investigated previously. AIM: To investigate the effects of glucosamine on insulin release or gene expression related to glucose metabolism in rat islets cultured with glucosamine for 24 hours. METHODOLOGY: After islets were cultured with glucosamine or diazoxide, we measured glucose- or arginine induced insulin release by using radioimmunoassay (RIA) and gene expressions by semiquantitative polymerase/chain reaction. RESULTS: Coculture with glucosamine inhibited 27 mmol/L glucose-induced insulin release with no effects on 20 mmol/L arginine-induced insulin release. Coculture with diazoxide did not restore the impaired glucose-induced insulin release. In glucosamine-cultured islets, glucose transporter type 2 or glucokinase mRNA expression decreased, whereas hexokinase mRNA increased. Phosphofructokinase-A, pyruvate dehydrogenase E1alpha, or pyruvate carboxylase mRNA was not affected by the addition of glucosamine. Pancreatic and duodenal homeobox-1, preproinsulin, or p21 (induced by oxidative stress) mRNA expression did not change, whereas uncoupling protein 2 mRNA, which plays an important role in thermogenesis, decreased in glucosamine-cultured islets. CONCLUSION: These data imply that glucosamine impairs glucose-induced insulin release probably through the inhibition of glucose metabolism. PMID- 11893930 TI - Reduction of immunogenicity by cyclophosphamide pretreatment and culture on xenograft survival of monkey pancreatic islets transplanted into rats. AB - INTRODUCTION: Islet cell transplantation offers a new approach for the treatment of diabetes. Before transplantation, immunomodulation procedures are conducted to reduce the immunogenicity of the pancreatic islets. AIM: To study the effect of cyclophosphamide pretreatment and culture on the xenograft survival of monkey pancreatic islets. METHODOLOGY: Islets were isolated from normal monkeys and transplanted under the renal capsule of the rats. The grafts were removed 7 days after transplantation and processed for histologic study to examine graft survival. Islets from normal monkeys without cyclosporine treatment were destroyed in 7 days. These islets required cyclosporine (30 mg/kg) to prevent rejection. In the second group, the donor monkeys were treated with cyclophosphamide (60 mg/kg body weight) -4 and -2 days before harvesting of the islets. On day 0, the islets were isolated and cultured in RPMI-1640 medium for 7 days in 95% air and 5% CO2. After culture, the islets were transplanted into the rats. The grafts were removed and processed for histologic study to examine graft survival. RESULTS: The pretreated and cultured islets required 15 mg/kg cyclosporine to prevent rejection. Half of the usual dose of cyclosporine is enough to prevent rejection if the donor monkeys were pretreated and the islets cultured. CONCLUSION: This study shows that donor pretreatment and culture reduces immunogenicity of the xenotransplanted primate islets into rats. PMID- 11893931 TI - A novel nude mouse model of liver metastasis and peritoneal dissemination from the same human pancreatic cancer line. AB - INTRODUCTION: Recently, several mice models have been used for investigating cancer metastasis. However, there are no metastatic and peritoneal dominated variants from the same parental cell line. AIM AND METHODOLOGY: To elucidate the mechanisms of metastasis, we established highly liver metastatic and peritoneal disseminated models in nude mice, and then characterized several factors related to metastasis in these cells. We established a series of well-characterized sublines that showed metastatic potentials to different organ sites of nude mice. Two sublines were selected sequentially from the parental pancreatic cancer cell line, HPC-4, resulting in a highly liver metastatic cell line, HPC-4H4, and a highly peritoneal disseminated cell line, HPC-4P4a. Using these three cell lines, we investigated several biologic properties and mRNA levels of differentially expressed genes involved in cancer metastasis. RESULTS: The tumorigenicity, the motile activity, and the adhesive activity of metastatic sublines were higher than those of parental HPC-4 cells. Macroscopic and microscopic findings and the DNA ploidy pattern were the same among the three cell lines. In addition, HPC-4H4 cells expressed clearly higher levels of vascular endothelial growth factor and IL-8 expression than did HPC-4P4a cells. In fluorescence-activated cell sorter analysis of adhesion molecules, the expression of integrin-alpha2 was enhanced in HPC-4 cells, integrin-alphavbeta5 was enhanced in HPC-4H4 cells, and integrin alpha3 was enhanced in HPC-4P4a cells. Osteopontin, vascular endothelial growth factor, and hepatocyte growth factor were among the genes that were upregulated in HPC-4H4 cells compared with HPC-4P4a cells. HPC-4P4a cells did not metastasize to the liver by intrasplenic injection. Conversely, HPC-4H4 cells metastasized remarkably to the peritoneum by intraabdominal injection. CONCLUSION: These sublines are the first reported liver metastatic and peritoneal disseminated models derived from the same parental cell lines. The results of our study suggest that the process of hematogenous metastasis is not the same as that of peritoneal dissemination. PMID- 11893932 TI - Expression of ROCK-1 in human pancreatic cancer: its down-regulation by morpholino oligo antisense can reduce the migration of pancreatic cancer cells in vitro. AB - INTRODUCTION: Invasion and metastasis of cancer cells require cell motility and adhesion. The small GTPase Rho and one of its effector molecules ROCK regulate cytoskeleton and actomyosin contractility, and play a crucial role in cell adhesion and motility. Results of previous studies showed that the elevated activity of ROCK-1, one of the isomers of ROCK kinases, led to an increase in the activity of invasion and metastasis of cancer cell lines. AIM: To investigate the importance of ROCK-1 in cancer invasion and metastasis. METHODOLOGY: We investigated the expression of ROCK-1 in two cancer cell lines and 31 human pancreatic tissues (21 pancreatic cancers [PC] and 10 histologically normal tissues) by immunoblotting and immunohistochemistry. We also examined by haptotaxis assay whether the migratory activity of PC cells could be suppressed by treatment with the morpholino antisense oligonucleotide in vitro. RESULTS: The expression of ROCK-1 was found in 18 of 21 PC tissues (85.7%), but not in normal pancreatic tissues by immunoblotting and immunohistochemistry. Antisense oligo against ROCK-1 significantly inhibited the haptotaxis of Panc-1 in a dose dependent manner, compared with the control oligo. CONCLUSION: These results suggest that ROCK-1 may contribute to pancreatic cancer cell invasion and/or metastasis by facilitating cancer cell migration. PMID- 11893933 TI - Emergency pancreatoduodenectomy in nontrauma patients. AB - INTRODUCTION: In recent years, the safety of pancreatoduodenectomy has improved, with a low mortality rate and reduced morbidity in institutions with extensive experience in the procedure. The indication for pancreatoduodenectomy has been expanded. Emergency pancreatoduodenectomy has mainly been performed for abdominal trauma. AIMS: To discuss possible indications for emergency pancreatoduodenectomy in nontrauma patients. METHODOLOGY: A series of 417 consecutive pancreatic head resections performed between November 1993 and August 2000 was reviewed for emergency interventions. Indications and outcome of emergency pancreatoduodenectomies were analyzed. RESULTS: The prevalence of emergency pancreatoduodenectomy was 1%. Two patients had duodenopancreatic complications after endoscopic and surgical interventions, and two patients had otherwise uncontrollable bleeding from a penetrating duodenal ulcer and an ampullary tumor. In all four patients, emergency pancreatoduodenectomy was carried out without local complications but with a high morbidity. One patient died after surgery. CONCLUSION: We conclude that emergency pancreatoduodenectomy may be considered, under exceptional circumstances, by surgeons experienced in pancreatic resections, but unfavorable perioperative conditions should be included in the preoperative planning and risk assessment of such patients. PMID- 11893934 TI - Penetration of meropenem and cefepim into pancreatic tissue during the course of experimental acute pancreatitis. AB - INTRODUCTION: Recent data from experimental and clinical studies suggest that the antibiotics showing good penetration into the pancreas may reduce mortality by preventing pancreatic infection, which is the most important prognostic factor in acute pancreatitis. AIM: To determine and compare pancreatic tissue concentrations of meropenem and cefepime at different stages of acute necrotizing pancreatitis in an animal model that has been shown to closely mimic severe human pancreatitis. METHODOLOGY: Acute necrotizing pancreatitis was induced in rats by a standardized intraductal infusion of glycodeoxycholic acid and intravenous cerulein. Six hours (n = 30) and 48 hours (n = 30) after induction of pancreatitis, the rats were randomized to receive an intravenous 20 mg/kg injection of either meropenem or cefepime. Blood and the head of the pancreas were collected for determining antibiotic concentrations by high-performance liquid chromatography. RESULTS: Meropenem concentrations in the pancreas at 6 hours of acute pancreatitis increased significantly and decreased at 48 hours of the disease, but were still higher than that in controls. Concentrations of cefepime in necrotic pancreatic tissue were significantly low either during the initial or later phase, but lower in latter, in which the necrosis was more evident. Tissue/serum concentration ratios of meropenem were significantly higher than those of cefepime. However, tissue concentrations of both antibiotics are much higher than the minimum inhibitory concentration values for the common microorganisms involved in pancreatic infections. CONCLUSION: Although both antibiotics penetrate into the necrotic tissue in sufficient therapeutic concentrations, penetration of meropenem is much better than cefepime. However, good tissue penetration may not solely indicate efficacy of that antibiotic. Therefore, further experimental and clinical studies are needed to determine the therapeutic and prognostic efficacy of these agents. PMID- 11893935 TI - Inhibitory effects of octreotide on luminal cholecystokinin-releasing factor, plasma cholecystokinin, and pancreatic secretion in conscious rats. AB - INTRODUCTION: Luminal cholecystokinin-releasing factor (LCRF), purified from rat intestinal secretions, is an intraluminal regulator of cholecystokinin (CCK) secretion during bile and pancreatic juice diversion. AIM: Because the LCRF content was not influenced by intravenous administration of atropine or somatostatin, the inhibitory effect of a potent somatostatin analog octreotide on LCRF content, the plasma CCK level, and pancreatic secretion was examined. METHODOLOGY: Rats were prepared with bile and pancreatic cannulae and two duodenal cannulae and with an external jugular vein cannula. After 1.5-hour basal collection, bile and pancreatic juice was diverted for 2 hours, during which octreotide was infused intravenously or into the duodenal lumen. The changes in pancreatic secretion were recorded for 2 hours, and the plasma CCK level and LCRF content 2 hours after the treatment were measured. RESULTS: Bile and pancreatic juice diversion significantly increased pancreatic secretion and plasma CCK and LCRF levels. Intravenous infusion of octreotide (0.2 and 0.5 nmol/kg/hour) inhibited all parameters. Intraduodenal infusion of a lower dose of octreotide (33 nmol/kg/hour) inhibited pancreatic secretion, but did not inhibit CCK release or LCRF content. The higher doses (100 and 300 nmol/kg/hour) inhibited all parameters. CONCLUSION: Intravenous and intraduodenal administrations of octreotide inhibited CCK release and LCRF content and pancreatic secretion induced by bile and pancreatic juice diversion. PMID- 11893937 TI - Parathyroid hormone-related protein as a novel tumor marker in pancreatic adenocarcinoma. AB - INTRODUCTION: Parathyroid hormone-related protein (PTHrP) can act as an oncoprotein to regulate the growth and proliferation of many common malignancies, including pancreatic cancer. Previous studies have shown that PTHrP is produced by human pancreatic cancer cell lines, can be shown in the cytoplasm and nucleus of paraffin-embedded pancreatic adenocarcinoma tumor specimens, and is secreted into the media of cultured pancreatic adenocarcinoma cells. We hypothesized that PTHrP could serve as a tumor-marker for growth of pancreatic cancer in vivo. AIM AND METHODOLOGY: To test this hypothesis, we used an orthotopic model developed in our laboratory of the PTHrP-producing human pancreatic cancer line, BxPC-3. This tumor was stably transduced with green fluorescence protein (GFP) to facilitate visualization of tumor growth and metastases. At early (5 weeks) and late (13 weeks) time points after surgical orthotopic implantation, serum PTHrP was measured and primary and metastatic tumor burden was determined for each mouse by assessing GFP expression. RESULTS: By 5 weeks after surgical orthotopic implantation (early group), the mean serum PTHrP level was 33.3 pg/mL. In contrast, by 13 weeks after surgical orthotopic implantation (late group), the mean serum PTHrP level increased to 158.5 pg/mL. These differences were highly significant (p < 0.001, Student t test). Numerous metastatic lesions were readily visualized by GFP in the late group. Serum PTHrP levels measured by immunoassay correlated with primary pancreatic tumor weights and serum calcium levels (p <0.01). PTHrP levels were not detectable (<21 pg/mL) in any of the 10 control mice with no tumor. Western blotting of BxPC-3-GFP tumor lysates confirmed the presence of PTHrP. BxPC-3-GFP tumor tissue stained with antibody to PTHrP. CONCLUSION: These results indicate that PTHrP can serve as a tumor marker in animal models of pancreatic cancer and may be a useful tumor marker for clinical pancreatic adenocarcinoma. PMID- 11893936 TI - Role of CCK-A receptor for pancreatic function in mice: a study in CCK-A receptor knockout mice. AB - INTRODUCTION: The cholecystokinin (CCK) family of peptides and receptors is present throughout the brain and gastrointestinal tract. The CCK receptors can be pharmacologically subdivided into two subtypes: CCK-A and CCK-B. CCK-A receptor is enriched in the pancreas of mice. AIMS: To determine pancreatic functions in a CCK-A receptor deficient mouse mutant generated by gene targeting in embryonic stem cells. The targeting vector contained lacZ and neo insertions in exon 2. METHODOLOGY: To examine exocrine functions, amylase release from the dispersed acini in vitro was examined. In the in vivo study, the mixture of bile-pancreatic juice was collected, and amylase, bicarbonate, and bile acid outputs were determined after the administration of various stimulants. The cystic duct of the gallbladder and the pylorus were ligated to exclude the involvement of gallbladder contraction and gastric acid. Pancreatic enzyme content was measured, and histologic examinations by HE and lacZ staining were conducted. To examine endocrine functions, oral glucose tolerance test (2 g/kg) was determined. RESULTS: The body weight, pancreatic wet weight, and enzyme content in the pancreas were similar among the three genotypes. Amylase release in vivo and in vitro and bicarbonate secretion in vivo were not stimulated by CCK-8 in CCK-AR ( /-) mice, whereas the responses to other stimulants were substantial in (-/-) mice. Administration of secretin did not increase bicarbonate secretion regardless of genotype. A normal glucose tolerance was observed in (-/-) mice. Acinar cells, islets, and duct cells were stained by lacZ, and HE staining revealed no pathologic findings. CONCLUSION: The CCK-A receptor is important for pancreatic exocrine secretion, but not essential for maintaining glucose concentration and pancreatic growth in mice. PMID- 11893938 TI - Differences in the expression of glutathione S-transferases in normal pancreas, chronic pancreatitis, secondary chronic pancreatitis, and pancreatic cancer. AB - INTRODUCTION: In our previous study, glutathione S-transferase-pi (GST-pi), a phase II drug metabolizing enzyme, was found to be expressed in pancreatic ductal and ductular cells but not acinar cells of the normal pancreas, chronic pancreatitis, and secondary pancreatitis caused by pancreatic cancer. A greater percentage of the cells expressing GST-pi was shown in the islets of chronic pancreatitis specimens compared with the normal pancreas and secondary pancreatitis. AIMS AND METHODOLOGY: To examine whether the increased number of islet cells expressing GST-pi and the absence in the acinar cells are compensated for by other GST isozymes, we investigated the expression of GST-alpha and GST-mu in the same specimens. RESULTS: Unlike the distribution of GST-pi, the distribution of GST-alpha and GST-mu in islets did not show marked differences between the three groups. However, in four of 18 primary chronic pancreatitis specimens, more islet cells (approximately 25%) expressed GST-alpha than in the normal pancreas and secondary chronic pancreatitis (both approximately 10%). The reactivity of cancer cells to GST-alpha, GST-mu, and GST-pi was similar to the ductal cells in the normal pancreas, chronic pancreatitis, and secondary chronic pancreatitis. Contrary to the expression of GST-pi, no statistically significant differences were found in the distribution of GST-alpha and GST-mu in the normal pancreas, chronic pancreatitis, and secondary chronic pancreatitis. CONCLUSION: The expression of the other GSTs does not compensate for the variation of expression of GST-pi. There was no specimen in each group that did not express at least one GST isozyme in islet, acinar, and ductal cells. PMID- 11893939 TI - Mast cells may not play a crucial role in the pathogenesis of experimental closed duodenal loop-induced pancreatitis in rats. AB - INTRODUCTION: Ws/Ws rats have a small deletion of the c-kit gene and are deficient in both mucosal-type mast cells and connective tissue-type mast cells. AIM: To investigate the role of pancreatic mast cells in the development of experimental closed duodenal loop (CDL)-induced pancreatitis using Ws/Ws rats. METHODOLOGY: Pancreatitis was induced by the CDL technique for 5 and 12 hours, and the subsequent ascites volume, wet pancreatic weight, pancreatic myeloperoxidase activities, and serum amylase levels were evaluated. The pancreatic tissue damage was also evaluated histologically. RESULTS: The CDL technique induced equally severe ascites, pancreatic edema and hyperemia, and hyperamylasemia in the Ws/Ws versus the control (+/+) rats. The microscopic mucosal damage score was also equivalent in the Ws/Ws and control (+/+) rats, and there were no significant differences in mucosal myeloperoxidase activity between the Ws/Ws and control (+/+) rats. CONCLUSION: These results indicate that mast cells may not be crucial for the development of CDL-induced pancreatitis. PMID- 11893941 TI - Intraductal papillary mucinous cystadenocarcinoma of the pancreas with intrapancreatic perineural invasion. PMID- 11893940 TI - Troglitazone stimulates pancreatic growth in normal rats. AB - INTRODUCTION: Peroxisome proliferator-activated receptors (PPARs) are a family of ligand-activated nuclear transcription factors belonging to the nuclear hormone receptors. Troglitazone, a specific ligand for PPAR-gamma is shown to regulate not only lipids and glucose metabolism, but also cell cycle, differentiation, and apoptosis. AIM: To examine the effect of chronic oral administration of troglitazone on the age-related changes of insulin resistance, plasma CCK levels, and pancreatic growth in normal rats. METHODOLOGY: A troglitazone-rich diet (0.2%) was given from 12 to 28 weeks of age or from 12 or 28 weeks of age to 72 weeks of age. RESULTS: Fasting serum glucose concentrations in control rats increased progressively with age, which was almost completely prevented by troglitazone treatment. Serum insulin concentrations and pancreatic insulin content in the control rat markedly increased at 28 weeks of age but decreased at 72 weeks of age. These parameters in troglitazone-treated rats remained at nearly the same concentrations at all ages. Insulin concentration relative to DNA in the control rats increased with age, whereas in the troglitazone-treated rats it remained at nearly the same concentrations throughout the observation periods and was significantly lower than that in the controls. Insulin resistance in control rats showed a great increase at 72 weeks of age, whereas it was nearly the same at all ages in troglitazone-treated rats and was significantly lower than those in the control rats. Plasma cholecystokinin concentrations in control rats slightly but insignificantly increased with age, whereas pancreatic weight decreased age-dependently when corrected for body weight. Although troglitazone treatment appeared to decrease plasma cholecystokinin concentrations compared with those in the control rats, it significantly increased pancreatic weight and prevented age-dependent decrease. Troglitazone treatment significantly increased pancreatic protein and DNA contents, but the protein per DNA ratio, an indicator of cellular size, remained at nearly the same concentrations at all ages. The contour of the islets in the control rats at 72 weeks of age was somewhat irregular with structural disarrangement and fibrosis. Moreover, the islets were separated into small sections (cluster) by fibrosis. Troglitazone treatment prevented or reversed these age-related changes of the islets to those in rats at 12 weeks of age. CONCLUSION: Our results indicate that troglitazone stimulates pancreatic growth in the normal rat not only by reducing insulin resistance and improving glucose metabolism, but also by suppressing fibrosis of the islets. PMID- 11893942 TI - The silent killer: fatal abscess as a late complication in chronic calcifying pancreatitis, painless for 11 years. PMID- 11893943 TI - Early changes of the expression of cathepsin B mRNA during cerulein supramaximal stimulation. PMID- 11893944 TI - Anterior segmentectomy of the pancreatic head for islet cell tumors. PMID- 11893946 TI - Hand washing for health-care workers in domestic care settings. AB - Hand washing is recognized as the single most important practice in reducing the spread of infection. Infection problems, such as an increase in antibiotic resistant organisms, have now made apparent the need to return to such basic practice. Although optimum facilities and practices for hand washing in an inpatient setting have been described, these are rarely available for health-care workers providing increasingly more complex care to patients and clients in their own homes. Therefore community staff need to adopt a risk assessment approach and strategies to ensure that adequate decontamination of their hands is achieved in order that care can be given with the minimal risk of cross-infection between patients, clients and staff. PMID- 11893945 TI - Health visiting under fire. Or is it? PMID- 11893947 TI - The case for researching the history of community nursing practice. AB - Despite a flurry of interest in the 1980s, the adoption of a positivist, objective, scientific paradigm for nursing research has led to a rejection of the study of nursing history as a valid pursuit in recent years. In this article, it is argued that this is a precarious situation. By not examining the history of the profession, nursing -- and in particular community nursing -- undermines its efforts to validate itself within the wider health-care arena. Nursing must learn from the mistakes of the past, as well as the successes, but do so in a critical way that does not romanticize its history. PMID- 11893948 TI - Pelvic floor reeducation for stress incontinence: comparing three methods. AB - Stress urinary incontinence is a common problem among women of all ages but may resolve with pelvic floor reeducation in many cases. Compliance to a regimen of pelvic floor muscle exercises is poor and many devices have been produced to make exercising these muscles more effective and interesting. This article describes a study in which two such devices -- vaginal cones and pressure biofeedback -- were compared with pelvic floor exercises alone. The results show that there is no statistically significant difference between the three modalities; all treatments produced significant improvement in symptoms and quality of life scores. PMID- 11893949 TI - Enuresis services: carers' knowledge and attitudes. AB - The research described in this article examines parents' and carers of children with bedwetting (enuresis) problems' attitudes to, and knowledge of, enuresis and enuresis services in southern Derbyshire. A specialist enuresis service was already running in this area and the continence nurse advisor (children's services) had noted a high number of referrals complaining about lack of knowledge prior to attending clinic and a lack of clarity and continuity of advice given by other health professionals. The specialist continence service needed to be needs-led and validated as an appropriate and adequate service. Following a literature search and pilot study, a questionnaire was sent to parents of children who had been referred to the clinic in the previous year. Analysis was performed using the SPSS statistical package. Results showed a high value being given to the specialist nurse-led service by parents and children and a high amount of conflicting, non-evidence based advice being given to parents and carers from various health professionals prior to referral to the specialist nurse. The results of the study led to the formulation of recommendations to improve services available to children with enuresis, their parents and carers. PMID- 11893950 TI - Nursing management of constipation in housebound older people. AB - In this article two algorithms are suggested for use by district nurses in the management of constipation in dependent older people at home. Prescribing for this group of patients requires a complex assessment of medical and social factors as 24-hour supervision is not always available and local health and social service resources may impact on the level of care that is available in terms of the provision of food and drink and the taking of medicines. PMID- 11893951 TI - Intermittent self-catheterisation in urinary tract reconstruction. PMID- 11893952 TI - Is a blueprint for care of older people enough? PMID- 11893953 TI - Nursing care is given too narrow a definition. PMID- 11893954 TI - Women's health in prison must be improved. PMID- 11893955 TI - Case 57; inappropriate relationships. Nurse who befriended intimately a teenage patient. PMID- 11893956 TI - Accurate selection of compression and antiembolic hosiery. AB - The use of compression and antiembolic hosiery is widespread in hospital and community settings, but there is wide variation in almost every aspect of selection and prescribing practice. In some localities pharmacists or appliance officers are responsible for selecting a suitable stocking for patients, whereas in others it may be left to medical staff. Anecdotally, there is a lack of knowledge about when and what is safe to prescribe relative to the underlying disorder. This is compounded by discrepancy between the level of compression provided by stockings made according to the European method, and that prescribable under the UK Drug Tariff in primary care. This can result in clinically serious variation in what the patient is given and what his/her condition needs. A particular problem arises when the patient's care is continued following discharge from hospital. In this case not only may an inferior quality product be supplied (meeting the Drug Tariff specifications), but also the compression 'strength' can be unintentionally lower than expected or prescribed. A simple method of prescribing according to condition and pressure has been developed and is outlined here. PMID- 11893957 TI - Poverty as a practice issue for learning disability nurses. AB - People who have a learning disability are vulnerable to poverty. Given that poverty has a negative impact on mental and physical health, this should be an area of concern for learning disability nurses. Historically, however, this is an area of practice which has not received much attention. It is important that learning disability nurses are aware of the impact which poverty can have on the lives of their clients and that they incorporate this understanding into their practice. Interventions which can be implemented at the level of individual clients, groups and society to tackle poverty and its effects are proposed. It is concluded that, at present, little is known about the extent to which such a perspective underpins practice or about the extent to which poverty is addressed in pre- and post- registration educational courses. The need for further research and development is noted. PMID- 11893958 TI - Down's syndrome: experiences of mothers from different cultures. AB - The purpose of this study is to explore the lived experiences of mothers from different cultural backgrounds who have a child with Down's syndrome. Today, in multicultural Britain, cultural awareness and understanding are necessary if relevant care is to be offered. People from different cultures have different health beliefs, understanding and expectations. In order to try to understand these differences, a small selected sample of mothers from three different cultures who had given birth to a child diagnosed with Down's syndrome were interviewed in their own homes. When their responses were analysed they demonstrated their comparative isolation and the influence of cultural beliefs, and also that despite much research and study there is still a lack of respect and value in the way they had been treated. The author of this article addresses the requirement of more sensitivity and empathy to parents' needs. PMID- 11893959 TI - Discussing cardiopulmonary resuscitation with patients. AB - The British Medical Association and the Royal College of Nursing acknowledge that patients should be involved in the decision-making process regarding cardiopulmonary resuscitation (CPR) in order to gain compliance with any decision made on their behalf. In the past, it was apparent that decisions concerning the eligibility of patients for CPR were being made arbitrarily and older people were being treated unfairly in respect of their consideration for this intervention. This article reviews literature associated with patient involvement in decision making regarding CPR and 'do not resuscitate' orders and how an assessment and rehabilitation unit promoted this activity as usual practice. In order to enhance partnership with patients and promote best possible practice in respect of CPR decision making the authors suggest a strategy that includes (1) evaluating documentation (2) development of a patient information leaflet and (3) an education programme for healthcare personnel. PMID- 11893960 TI - Presentation skills for nurses: how to prepare more effectively. AB - Many clinical nurses are changing the way they deliver care for the benefit of their patients. In order to share ideas, knowledge and opinions more widely, nurses need to either speak or write within the public arena. For all nurses, presentation skills are not just useful, they are essential, whether for sharing practice, influencing colleagues or seeking a new job. Such skills can be learnt, building on characteristics most nurses already possess. Effective presentations usually require learning to control nervousness and preparing properly. This article provides a framework for preparation and suggestions for ways to achieve calm and confidence. PMID- 11893961 TI - Legal aspects of consent 14: organ removal, retention, storage. AB - Case Scenario: Sarah was born with a congenital heart condition. Unfortunately, the subsequent operation proved unsuccessful and Sarah died. Sarah's parents were asked if they would agree to a post mortem being performed to assist in research so that in future such conditions could be successfully operated upon. The parents agreed and subsequently they were notified that the body was available for disposal. They decided upon a cremation. Several years later, following an inquiry into the pathology services of the hospital, they were notified by the Chief Executive's department that Sarah's heart, lungs, liver and other organs had been retained. The parents were shocked. What is the law? PMID- 11893962 TI - Hospital league tables: a help or hindrance? PMID- 11893963 TI - Pulmonary embolism diagnosis. What role for nuclear medicine. PMID- 11893964 TI - Ventilation/perfusion scan and dead space in pulmonary embolism: are they useful for the diagnosis? AB - The diagnostic strategy for pulmonary embolism, based on the mismatch of the ventilation/perfusion scan, was developed some 30 years ago on the following assumption: since the disorder involves the pulmonary vessels, it was surmised that in the embolized regions lung alveoli are unperfused or poorly perfused but well ventilated. Hence, it was inferred that this disorder was characterized, unlike parenchymal disease, by ventilation/perfusion mismatch in the affected lung zones and by an obvious increase of wasted ventilation, i.e., dead space. As matter of fact, experimental evidence on the redistribution of ventilation away from the vascular occluded lung had been already obtained in the early 60s of the last century. More recently, the behavior of regional pulmonary ventilation (V(A)) and blood flow (Q) in patients with acute pulmonary embolism (APE) has been studied by applying the multiple inert gas elimination technique (MIGET). It has been shown that the development of lung units with high V(A)/Q ratio (those with relative prevalence of perfusion obstruction) is accompanied by substantial redistribution of ventilation away from these units. Furthermore, radioisotopic techniques, used to visualize the topographic distributions of V(A) and Q in the same patients studied by MIGET, have shown reduced or absent V(A) in the embolized regions. This may occur by different mechanisms in the various stages of APE: bronchoconstriction mediated by local hypocapnia, atelectasis (occasionally hemorrhagic) related to alteration of surfactant production, bronchiolar obstruction and pulmonary infarction ascribed to degenerative and/or necrotic changes secondary to insufficient blood flow. In dogs and humans alike, the dead space measured by MIGET does not increase and that obtained from CO2 increases far less than the amount of unperfused lung in APE thus confirming a substantial redistribution of ventilation away from the embolized lung zones. Taken together, all these observations provide the pathophysiological explanation of the unacceptedly low level of sensitivity for the diagnostic strategy of APE based on the mismatch of the ventilation/perfusion scan. PMID- 11893965 TI - Assessing the clinical probability of pulmonary embolism. AB - Clinical assessment is a cornerstone of the recently validated diagnostic strategies for pulmonary embolism (PE). Although the diagnostic yield of individual symptoms, signs, and common laboratory tests is limited, the combination of these variables, either by empirical assessment or by a prediction rule, can be used to express a clinical probability of PE. The latter may serve as pretest probability to predict the probability of PE after further objective testing (posterior or post-test probability). Over the last few years, attempts have been made to develop structured prediction models for PE. In a Canadian multicenter prospective study, the clinical probability of PE was rated as low, intermediate, or high according to a model which included assessment of presenting symptoms and signs, risk factors, and presence or absence of an alternative diagnosis at least as likely as PE. The prevalence of PE in the low, intermediate, and high pretest probability categories was 3, 28, and 78%, respectively. This model relies heavily on the clinician's subjective judgement as to whether an alternative diagnosis is as likely as or more likely than PE, and, as such, it can be hardly standardized. Furthermore, the inherent complexity of the model may limit its applicability in daily clinical practice. Recently, a simple clinical score was developed to stratify outpatients with suspected PE into groups with low, intermediate, or high clinical probability. Logistic regression was used to predict parameters associated with PE. A score =/<4 identified patients with low probability of whom 10% had PE. The prevalence of PE in patients with intermediate (score 5-8) and high probability (score > or = 9) was 38 and 81%, respectively. As opposed to the Canadian model, this clinical score is standardized. The predictor variables identified in the model, however, were derived from a data base of emergency ward patients. This model may, therefore, not be valid in assessing the clinical probability of PE in inpatients. In the PISA-PED study, a clinical diagnostic algorithm was developed which rests on the identification of three relevant clinical symptoms and on their association with electrocardiographic and/or radiographic abnormalities specific for PE. Among patients who, according to the model, had been rated as having a high clinical probability, the prevalence of proven PE was 97%, while it was 3% in those with low probability. The prevalence of PE in patients with intermediate clinical probability was 41%. These results underscore the importance of incorporating the standardized reading of the electrocardiogram and of the chest radiograph into the clinical evaluation of patients with suspected PE. The interpretation of these laboratory data, however, requires experience. Future research is needed to develop standardized models, of varying degree of complexity, which may find application in different clinical settings to predict the probability of PE. PMID- 11893966 TI - Current role of lung scintigraphy in pulmonary embolism. AB - The pivotal role of lung scintigraphy in the diagnosis of pulmonary embolism (PE) has been questioned in recent years due to the introduction of spiral computed tomography. However, the scintigraphic results used for comparisons are often those of the authoritative PIOPED (Prospective Investigation of Pulmonary Embolism Diagnosis) study, carried out in the 1980s. Pulmonary scintigraphy has progressed from those years both in the methodological and interpretative fields, although perhaps too slowly. Results better than those of PIOPED's have been presented by study groups who used: 1) perfusion-only approach, 2) SPET imaging; 3) new interpretative criteria; 4) different prediction rules to integrate clinical and scintigraphic probabilities of PE. These advances are still insufficiently recognised by the nuclear medicine community, possibly due to a sort of PIOPED-based "cultural globalisation". This paper reviews the actual advantages and limitations of nuclear medicine techniques, the diagnostic role of scintigraphy within the diagnostic algorithms proposed by international working groups and scientific societies and the results obtained from SPET imaging in the diagnosis of PE. PMID- 11893967 TI - Venous thromboembolic disease: CT evaluation. AB - Helical and multidetector CT has proven to be a valuable imaging modality for both pulmonary embolism and deep venous thrombosis. This paper will review the sensitivity and specificity of CT and discuss diagnostic algorithms utilizing CT and more established imaging technologies. PMID- 11893968 TI - MRA for diagnosis of venous thromboembolism. AB - Direct imaging of pulmonary embolism (PE) and deep vein thrombosis (DVT) with CT, and potentially with MR, will continue to replace V/Q scintigraphy. Venous imaging with MR far detecting DVT is used in a few centers, and their published accuracy figures are impressive. Recent studies of MR pulmonary angiography for PE reported that sensitivity of MRA was 85-100%, specificity 95-96%, but this data must be confirmed in other centers and patient populations. MR has advantages compared with CT, which make it worthwhile to continue MR development. Ionizing radiation and iodinated contrast material are not used. Imaging the pulmonary arteries and then imaging whichever venous region is of clinical interest is practical in a single examination. Repeated examinations can be performed safely. New contrast materials will facilitate the practicality and accuracy of the MR technique and perfusion imaging may increase sensitivity. MR also has disadvantages compared with CT. It does not image effectively the non vascular compartment of the lungs. It is more expensive, patient monitoring is more cumbersome, and a routine technique, which embodies all of MR's potential advantages, has not been packaged and tested. Accordingly, helical CT is a realistic option in clinical management of patients with suspected PE in most centers, while clinical application of MR is limited to centers with appropriate MR expertise and technology. However, MR has a number of fundamental characteristics that make it a potentially ideal modality for evaluating patients with suspected acute venous thromboembolic disease and further clinical research with MRA is warranted. PMID- 11893969 TI - Diagnosis of deep venous thrombosis. A review of radiologic, radionuclide, and non-imaging methods. AB - Accurate diagnosis of deep venous thrombosis (DVT) can be achieved through appropriate use of the various imaging and non-imaging techniques currently available in clinical practice. This paper summarizes the roles of imaging techniques such as duplex ultrasound, contrast and radionuclide venography, and magnetic resonance imaging, as well as clinical prediction models and D-dimer testing, in the evaluation of patients with suspected DVT. Recent data examining the prognostic value of several of these tests, alone and in combination, are also reviewed. The associations of testing for DVT and pulmonary embolism, and the controversies which sometimes surround them, are also briefly examined. PMID- 11893971 TI - Acidic drinks and referred otalgia. PMID- 11893973 TI - Effectiveness of perioperative bupivacaine infiltration in tonsillectomy patients. AB - PURPOSE: To determine the effectiveness of perioperative local anesthesia in tonsillectomy patients. DESIGN: Two-group prospective, blinded, randomized study. Patients in one group received a local infiltration of 10 mL 0.5% bupivacaine solution, and in the other group they were infiltrated with 10 mL of normal saline. RESULTS: Thirty-four adults agreed to participate in the study over a 2 year period, with 20 patients returning completed questionnaire forms. Regarding pain intensity over a 10-day period, oral analgesic use after surgery, and oral intake after surgery, there were no differences between the two groups, with P values of .25, .32, and .34, respectively. CONCLUSION: The effect of perioperative local marcain infiltration in tonsillectomy patients was similar to placebo. PMID- 11893972 TI - Nasal and otologic effects of experimental respiratory syncytial virus infection in adults. AB - PURPOSE: Upper respiratory tract viral infections continue to cause substantial patient morbidity and complications including sinusitis, otitis media, and pneumonia. This study was conducted to more clearly define the extent and frequency of nasal and otologic effects of respiratory syncytial virus infection in healthy adult volunteers. MATERIALS AND METHODS: Thirty-two healthy, susceptible adult volunteers were cloistered for a 9-day period. During this time, subjects underwent nasal inoculation with respiratory syncytial virus. Monitoring included daily self-assessment of general health, as well as nasal and otologic symptoms. Objective measurements of nasal and otologic function included expelled nasal secretion weight, saccharin-dye mucociliary clearance, sonotubometry, tympanometry, and physical examination. A nasal lavage was done each morning, and fluids were submitted for viral detection. Twenty-one days after viral inoculation, all subjects had convalescent blood samples drawn for assay of virus-specific antibody titres. RESULTS: Eighteen subjects (56%) became infected with the challenge as determined by either viral shedding (47%) or antigen detection (41%) from the nose or a 4-fold rise in virus-specific antibody titer (34%). Infected subjects more frequently reported adverse nasal (congestion, rhinorrhea) and general symptoms (fever, malaise, illness). By day 6, only 46% of infected subjects had normal middle ear pressure (> -100 mm H2O). Nasal secretion also increased substantially after infection. No patterned changes in mucociliary clearance or sonotubometry occurred, and no subjects developed otitis media. CONCLUSIONS: Nasal inoculation of healthy, susceptible adult volunteers with respiratory syncytial virus results in detectable infection in only about half of the subjects challenged. Infected subjects experience substantial symptoms and signs of a viral upper respiratory tract illness. As in our previous studies using rhinovirus and influenza A virus, respiratory syncytial virus disrupted the maintenance of normal middle ear pressures. These findings provide further support for the relationship between viral upper respiratory tract infections and otitis media. PMID- 11893975 TI - Septoplasty: long-term evaluation of results. AB - OBJECTIVE: The goal of this study was to ascertain how accurately the surgeons' selection criteria for septoplasty, which largely relies on clinical judgement alone, is able to anticipate patients' long-term satisfaction. STUDY DESIGN AND SETTING: A 10-year retrospective study was undertaken in a tertiary care otorhinolaryngology center. METHODS: All patients who underwent septoplasty at the same otorhinolaryngology department in the past 2 to 10 years were mailed a questionnaire to evaluate their perception of the procedure's results. Clinical files were reviewed to establish the initial type of septal deformity. Because some surgeons used rhinomanometry to support their decision to recommend surgery, its capability of predicting patients' satisfaction was simultaneously assessed. RESULTS: Subjects with anterior septal deformities were shown to benefit the most from septoplasty. Selection of patients on clinical grounds alone does carry, however, a considerable risk of patient dissatisfaction with end results. Nevertheless, despite a very strong correlation between anterior septal deviations and increased nasal resistance, preoperative rhinomanometry data failed to prove useful in predicting the long-term surgical outcome. CONCLUSIONS: Surgeons' appreciation of the types of septal deviation that do benefit from surgical correction falls short of desirable. Patients' satisfaction, however, did not improve if rhinomanometry was included in the preoperative evaluation. PMID- 11893974 TI - Reinke's edema and risk factors: clinical and histopathologic aspects. AB - PURPOSE: The purpose of this study was to evaluate retrospectively the distribution of histologic damage and its correlation with various risk factors in a group of patients affected by Reinke's edema. MATERIALS AND METHODS: The study subjects comprised 125 patients with bilateral Reinke's edema consecutively presenting at the Department of Otorhinolaryngology, "La Sapienza" University, Rome. The patients were divided into 4 groups according to Hellquist, Lundgren, and Olofsson's histologic classification and were then further categorized according to the number of cigarettes they smoked daily. Average exposure to cigarette smoke, occupation, habitual voice use, and gastroesophageal reflux were also considered. RESULTS: Fifty-two patients did not exhibit histologic lesions, 64 patients were histologically classified as Group 1 (epithelial hyperplasia and/or keratosis with or without mild dysplasia), and 6 patients exhibited moderate dysplasia (Group 2). In 3 patients, histologic examination showed evidence of unilateral carcinoma in situ (Group 3). Forty-four patients suffered recurrences within the first 2.5 years. Both daily cigarette consumption and duration of exposure to cigarette smoke were found to influence the severity of the histologic lesion. An association with gastroesophageal reflux was observed in 4 patients (3.2%). Prolonged vocal abuse did not prove to be a noteworthy factor in our study. CONCLUSIONS: The main risk factor for Reinke's edema and for its recurrence is tobacco use. Our study results showed that the clinical manifestation of this disease is related to the number of cigarettes smoked daily and the duration of exposure to smoke. Longer durations of exposure to cigarette smoke result in higher degrees of histologic damage. PMID- 11893976 TI - Leech in the epiglottis: an unusual discovery in our times. PMID- 11893977 TI - Follicular dendritic cell tumor: an aggressive head and neck tumor. AB - OBJECTIVE: To characterize the clinicopathologic features of head and neck follicular dendritic cell (FDC) tumor and report the experience of this entity at our institution. STUDY DESIGN: Two case presentations are compared with a retrospective analysis of all published head and neck cases. SETTING: A tertiary academic medical center. RESULTS: Thirty four cases of FDC tumor of the head and neck cases have been published. Twenty five occurred in the cervical lymph nodes, 4 in the tonsils, 2 in the palate, 1 in the pharynx, 1 in the parapharyngeal region, and 1 in the thyroid gland. Patient ages ranged from 13 to 73 years (mean, 38), and there was a roughly equal number of men and women. Patients were treated with surgery (17), surgery and chemotherapy (8), and surgery and radiation (9). After the primary treatment, 12 patients had no evidence of disease, whereas 5 were incurable. Ten tumors recurred locally and 3 distally. Of these 13 patients who suffered recurrences, 4 had no evidence of disease after secondary treatment, 6 were alive with disease, and one was lost to follow up. Two patients died after recurrence. We add 2 unique cases to the 9 previously reported extranodal cases, 1 in the tonsil and 1 in the parotid gland. CONCLUSION: FDC tumor is a rare malignant neoplasm that can present in the head and neck region in both lymph nodes and extranodal sites. Because of their rarity, these tumors are probably underrecognized by both clinicians and pathologists. Distinct light microscopic, immunohistochemical, and ultrastructural features do exist, however, and are reviewed. Surgery has been the mainstay of treatment and should include diligent control of surgical margins. The role of adjuvant therapy remains controversial. Although originally considered to be a low-grade malignancy, our review suggests both high recurrence rates and metastatic potential. We believe that FDC tumor should be viewed and treated as a moderately aggressive head and neck tumor. PMID- 11893978 TI - Otorhinolaryngologic manifestations in Chiari malformation. AB - The Chiari malformation causes herniation of the cerebellar amygdalae through the foramen magnum, resulting in the descent of the brain stem and/or traction on the lower cranial pairs. It is important for otolaryngologists to recognize Chiari malformations as part of the differential diagnosis of balance disorders, because patients may initially exhibit symptoms related to the vestibular system, including ataxia, nystagmus, or vertigo. We report 2 cases. PMID- 11893979 TI - Recurrent ameloblastoma presenting in the temporal fossa. AB - Recurrent ameloblastoma presenting in the temporal fossa is a rarely reported clinical entity. This report presents a case of temporal recurrence that occurred 30 years after the initial surgical treatment. A 45-year-old woman had a history of multiple enucleations for recurrences of her right mandibular ameloblastoma. The tumor was finally resected and reconstructed with an autogenous rib graft, after which the lesion reappeared in the temporal fossa 10 years later. We hypothesized that this temporal ameloblastoma is an implantation lesion seeded via the intraoral wound during the previous resection and reconstruction. PMID- 11893980 TI - Leiomyosarcoma of the sinonasal tract: a case report and literature review. AB - Leiomyosarcoma of the paranasal sinuses is uncommon and has an aggressive clinical behavior. Only 28 cases have been described in the literature, and, of these, only 3 patients treated with surgery had a disease-free survival. In this report, we describe the clinical history of a 57-year-old woman with a leiomyosarcoma of the paranasal sinuses successfully treated with a combination of surgery, radiation therapy, and chemotherapy. We obtained a total response with ifosfamide, doxorubicin, dacarbazine, and epirubicin. This case report emphasizes the possibility of treating this aggressive tumor with chemotherapy in accordance with the same therapy used in the treatment of uterine leiomyosarcomas. PMID- 11893981 TI - Subglottic plasmacytoma: a case report and review of the literature. PMID- 11893982 TI - Parotid abscess caused by Salmonella enteriditis in a patient with parotid masses. PMID- 11893983 TI - Electrical therapy in patients with congestive heart failure introduction. PMID- 11893984 TI - Measuring social anxiety and obsessive-compulsive spectra: comparison of interviews and self-report instruments. AB - The present report analyzes the agreement between the interview and the self report formats of the instruments Structured Clinical Interview for Social Anxiety Spectrum (SCI-SHY) and Structured Clinical Interview for Obsessive Compulsive Spectrum (SCI-OBS), already validated, in three psychiatric patient samples and controls. Thirty patients (10 with obsessive-compulsive disorder [OCD], 10 with social anxiety disorder [SAD], 10 with recurrent unipolar depression in remission) and 20 control subjects (10 university students, 10 ophthalmologic patients) were assessed using the SCI-SHY, the SCI-OBS, and the self report version of the two instruments. Agreement between the two versions was very good for the seven SCI-OBS domains (with intraclass correlation coefficients [ICCs] ranging from 0.80 to 0.96) and the four SCI-SHY domains (ICCs from 0.74 to 0.90). When items were analyzed individually, subjects tended to under-report some phobia-related problems in the interview. The total number of items endorsed in the SCI-SHY, but not in the SCI-OBS, was affected by the order of administration: when the SCI-SHY interview was administered first, subjects reported a median of five more symptoms; when the self-report was administered first, there was no significant difference in the number of symptoms endorsed in the two formats. However, this difference is not clinically important, given the large number of items comprising the instruments, and might be explained by the fact that subjects are likely to overemphasize occasional symptoms or behaviors when they are asked by the interviewer to answer a long series of "new" questions as accurately as possible. Given the high agreement between domain scores in the two formats of the instruments and the fact that scores are virtually identical when the self-report is administered first, we recommend the use of the self report versions in clinical and research settings. PMID- 11893986 TI - Clinical and research measures of grief: a reconsideration. AB - Bereavement-induced grief and psychological intervention are important social issues and worthy of greater attention from researchers and clinicians. Here, we review currently available measures of grief and discusses the differentiation of normal grief reaction from pathological grief and major depression. Finally, we propose future directions for research on the development of new grief measures and the effects of normal and pathological grief on psychological and physical health. PMID- 11893985 TI - Predictors of entering a long-term drug treatment study of panic disorder. AB - Three hundred thirty-three consecutive individuals with panic and agoraphobic like symptoms were evaluated as part of the screening for entry in a long-term imipramine treatment study of panic disorder with agoraphobia. The present report compares three subgroups of potential subjects--those who entered the study (n = 139, 41.7%), those who were rejected from participation based on exclusion criteria (n = 161, 48.3%), and those who qualified to enter but refused participation (n = 33, 10.0%)-in order to characterize pretreatment attrition and address two specific questions: What variables other than inclusion and exclusion criteria predict the likelihood of entering the trial?, and What variables significantly predict the likelihood of refusal to participate? Logistic regression analysis revealed that the likelihood of entering the trial was influenced significantly and independently by a number of variables that included extroversion, anxiety, and work satisfaction and performance measures. The refused and rejected groups differed significantly on a number of variables that were not a priori exclusionary criteria, such as a history of substance abuse other than alcohol and current use of alcohol to decrease anxiety, but none of these variables emerged as independent, significant predictors of refusing behavior as a separate category of nonparticipation. Findings suggest the presence of factors common to both the rejected and the refused groups that significantly influence pretreatment attrition and caution that categorizing pretreatment attrition into refused and rejected categories may not always be clear cut. Research and clinical implications are briefly discussed. PMID- 11893987 TI - Predictors of outcome in a 27-year follow-up of patients with borderline personality disorder. AB - Sixty-four patients with borderline personality disorder (BPD) were followed up for a mean of 27 years. Outcome was assessed using the Diagnostic Interview for Borderlines, Revised (DIB-R), the Global Assessment of Functioning (GAF), the Symptom Check List-90 (SCL-90), and the Social Adjustment Scale (SAS-SR). Subjects were also administered two self-report measures of childhood experience: the Parental Bonding Index (PBI) and the Developmental Experiences Questionnaire (DEQ). DIB-R scores and GAF scores at 15 years were significant predictors of all these measures of long-term outcome. In contrast, reports of parenting quality and of childhood abuse or trauma had no relationship to outcome. PMID- 11893988 TI - Teasing history and eating disorder features: an age- and body mass index-matched comparison of bulimia nervosa and binge-eating disorder. AB - The study examined whether a history of being teased about physical appearance is associated with differential patterns of current symptomatology in patients with bulimia nervosa (BN) as compared to patients with binge-eating disorder (BED). Thirty-two adult female patients with BN were compared with an age- and body mass index (BMI)-matched group of 32 adult female patients with BED. A battery of established instruments were used to assess physical appearance-related teasing history, current eating disorder features, body dissatisfaction, and psychological functioning (depression and self-esteem). BN patients reported a significantly higher frequency of having been teased about weight and size (WST) but a similar frequency of having been teased about general appearance (GAT) as the age- and BMI-matched BED patients. The two study groups did not differ significantly on current eating disorder features, body dissatisfaction, or psychological functioning, with the one exception that BN patients were characterized by significantly greater dietary restraint. Correlational analyses conducted separately within the BN and BED study groups revealed few significant associations between teasing history and eating disorder features in either study group. For the BN group, neither WST nor GAT was significantly associated with eating disorder features or body dissatisfaction, but both were significantly associated with lower self-esteem. For the age- and BMI-matched BED group, WST was not associated with eating disorder features, body dissatisfaction, or psychological functioning, but GAT was associated with higher dietary restraint and depression. Although physical appearance related teasing history was not associated with most eating disorder features for either study group, it appears that the association of different forms of teasing with psychological functioning may vary depending on the type of eating disorder symptomatology. PMID- 11893989 TI - Anorectic family dynamics: temperament and character data. AB - Modern psychobiologic research conceptualizes personality as a complex adaptive system involving a bidirectional interaction between heritable neurobiologic dispositions (temperament) and social learning (character). In this study, we evaluated temperament and character traits of patients with anorexia nervosa and their mothers and fathers, and we analyzed the correlation of temperament and character traits among family members in anorectic families. Finally, we tested the ability of the Temperament and Character Inventory (TCI) to discriminate between normal controls and anorectic subjects, their parents, and their families. Temperament and character features of 50 restricter anorectic patients and their parents (23 fathers and 25 mothers) were analyzed and then compared with a control group of 60 women and their 20 fathers and 20 mothers using the TCI. Data suggest that both temperament and character factors are involved in anorexia nervosa (AN). Anorectic individuals were high in harm avoidance (HA), low in novelty seeking (NS), and high in persistence (P) ("obsessive temperament type"). Their character was remarkable for low self-directedness (SD). Their mothers were distinguished by low SD. The fathers were high in HA, but also low in P, and high in reward dependence (RD). Again, they were low in SD. The anorectic family had low SD as a common denominator observed in all family members. This finding indicates that the psychopathology of AN extends beyond obsessiveness, but combines obsessiveness with low character development. None of the above temperament and character profiles is pathognomic of restricter anorectics. The observation that both temperament and character have an important role in the etiopathogenesis of AN has important treatment ramifications. The TCI was useful in discriminating between normal controls and anorectic subjects, their parents, and the whole anorectic family. PMID- 11893991 TI - Impulsive personality traits in male pedophiles versus healthy controls: is pedophilia an impulsive-aggressive disorder? AB - Pedophilia is characterized by sexual attraction to prepubescent children. Despite the extensive literature documenting the pervasive and pernicious effects of childhood sexual abuse, there is surprisingly little psychiatric literature on pedophilia and its etiology remains enigmatic. In recent years, the psychiatric literature on the phenomenology, neurobiology, and treatment of impulsive aggressive disorders has grown significantly. As some investigators have conceptualized pedophilia as an impulsive-aggressive disorder, it is of interest whether recent advances in the study of impulsive-aggressive disorders might shed light on pathological mechanisms underlying pedophilia. In the following study, 20 male subjects with a DSM-IV diagnosis of pedophilia, heterosexual type were recruited from an outpatient facility for sexual offenders and compared to 24 demographically similar control subjects. Groups were compared on three personality instruments--the Millon Clinical Multiaxial Inventory-II (MCMI-II), the Temperament and Character Inventory (TCI), and the Dimensional Assessment of Personality Impairment-Questionnaire (DAPI-Q)--to assess for select impairment in impulsive-aggressive personality traits. Pedophiles showed severe and pervasive personality impairment relative to controls. Although there was evidence of impulsivity, the findings do not suggest a predominance of impulsive-aggressive traits, and in fact provide evidence of inhibition, passive-aggression, and harm avoidance. The notion of "compulsive-aggression" in pedophilia is proposed. PMID- 11893990 TI - Personality profiles of patients with dissociative trance disorder in Singapore. AB - This report studies the personality of individuals who suffer from dissociative trance disorder and examines whether the personality profiles could predict the individual's frequency of trance states. A total of 58 cases were given the Eysenck Personality Questionnaire (EPQ) and their personal information harvested from the case notes and through subsequent interviews. The subjects were also reassessed 1 year later to obtain their frequency of trance states. For both sexes, there were lower extraversion scores and higher psychoticism, neuroticism, and lie scores in the sample compared to the Singapore norms. Of the 47 subjects traced, total episodes of trances that occurred over the 1-year period was positively correlated with neuroticism and negatively with extraversion scores. The high lie scores in individuals with dissociative trance disorder could be a reflection of their concern of how others perceive them. The motivation could be that of restoration of self-esteem or "face." The profiles in the EPQ could be used to predict the individual's frequency of trance states. Subjects with personality traits like nervousness, excitability, and emotional instability were more likely to have a higher frequency of trance states. PMID- 11893992 TI - Neuropsychiatric systemic lupus erythematosus, age, and the neurodevelopmental model: evidence in support of the Weinberger hypothesis. AB - We set out to describe neurologic and psychiatric syndromes secondary to systemic lupus erythematosus (NPSLE) in terms of age of onset and to compare such findings to those predicted by Weinberger's "windows of vulnerability" hypothesis of the neurodevelopmental pathogenesis of mental disorders. A search of the literature provided 767 NPSLE diagnoses in 511 patients. The particular NPSLE diagnoses and the age at which they occurred were compiled. We then determined ranges, medians, and means of age of onset of the various syndromes. We compared these to the ranges and means of age of onset of psychiatric disorders postulated in the "windows" model. The type of NPSLE varied with age and the most striking finding was an 11.3-year difference between the mean onset of psychosis (30.4) and depressed mood (41.1). The mean ages of onset and the ranges were similar to those proposed by Weinberger both absolutely and in temporal relation to each other. We conclude that NPSLE syndromes vary in age of onset in a manner consistent with, and in support of, the "windows of vulnerability" neurodevelopmental model of mental disorders. PMID- 11893993 TI - Predicting recidivism in delinquent adolescents from psychological and psychiatric assessment. AB - An important but sparse body of research focuses on psychiatric disorders and/or psychological characteristics as predictors of recidivism in delinquent adolescents. Through detailed psychiatric and psychological assessment in 64 adjudicated male juvenile delinquents, the current study investigated the long term relationships between psychopathology, intellectual functioning, and recidivism. Psychiatric diagnosis was based on the results of a semistructured interview conducted both with delinquents and their parents. After 2 years, information on recidivism was retrieved from the official records for the 64 subjects who underwent an initial assessment and the 36 who did not but who were adjudicated during the same period. The statistical model of future offending was able to explain 44.0% of the variance through the combination of (1) diagnosis of conduct disorder, (2) absence of major depression, and (3) low verbal IQ as scored on a standard intelligence test. It is concluded that specific aspects of psychiatric and psychological assessment can help to predict recidivist offending in delinquent adolescents. Future research should focus on the possibilities of implementing these findings in the development of prevention and rehabilitation programs. PMID- 11893994 TI - Obsessive-compulsive disorder with poor insight. AB - Although a diagnosis of obsessive-compulsive disorder (OCD) can be made with the specification "poor insight" (PI), this subtype remains understudied. To investigate the subtype, 78 OCD patients were characterized by degree of insight, reevaluated after treatment, and compared with 20 schizophrenics with OCD (OCD+S). At the pretreatment assessments in OCD patients, 28 subjects with poor or delusional insight (PI; 36%) were distinguished from 50 subjects with fair or good insight (GI; 64%) using the insight question of the Yale-Brown Obsessive Compulsive Scale (Y-BOCS). Compared to the OCD+S group, OCD patients were less likely to have PI, whereas OCD PI patients showed a similar degree of functional impairment as that observed in the OCD+S. After a 6-month combination of clomipramine with cognitive-behavioral treatment, 14 of 25 OCD PI patients no longer fell in the PI category, which was associated with reduced OCD severity and depressive status. Schizotypal personality disorder (SPD) was more common in patients whose insight remained poor even after the treatment. OCD patients demonstrate a range of insight with PI accompanied by significant dysfunction. Comorbid SPD in PI patients may be associated with worse prognosis. PMID- 11893996 TI - Two landmark achievements. PMID- 11893995 TI - Cloninger's Tridimensional Personality Questionnaire: psychometric properties and construct validity in Taiwanese adults. AB - Despite the wide use of the Tridimensional Personality Questionnaire (TPQ) in Western populations as a useful tool integrating both genetic and environmental influences on personality, some of its constructs remain questionable. In this study, we examined the psychometric properties of the TPQ and its relationship with aggression in Taiwanese adults. The subjects were 201 Taiwanese adults of wide ranges in age and educational level. Subjects were assessed using a Chinese version of the TPQ and the Brown-Goodwin Aggression Inventory. The internal consistency of the Chinese version of the TPQ scales is found to be mostly in the acceptable range except for the reward dependence (RD) scale and its subscales. The results of factor analysis of the 12 TPQ subscales partially support the four factor model rather than the original three-factor model. The construct validity of the novelty seeking (NS) and harm avoidance (HA) dimensions is supported by the findings that the NS is negatively correlated with age, the NS1 subscale is slightly negatively correlated with all the HA subscales, and the NS is positively correlated with aggression. We conclude that both the HA and NS scales of the TPQ are cross-culturally robust, while the RD scale needs to be refined. PMID- 11893997 TI - The American Society of Pain Management Nurses practice analysis: role delineation study. AB - A role delineation study was carried out by the American Society of Pain Management Nurses (ASPMN) to determine the activities performed by nurses involved in pain management. Demographic characteristics of pain management nurses and differences in practice based on educational preparation and specialty affiliation were identified. The role delineation survey was developed based on Nursing Intervention Classifications. The survey consisted of 91 activity statements. Respondents rated the frequency of performing each activity and the importance of the activity. A Mean Activity Index score was calculated for each item. All members of the ASPMN, the nursing special interest group (SIG) of the American Pain Society, and the pain SIG of the Oncology Nursing Society, as well as random samples of members of the National Association of Orthopaedic Nurses, Hospice and Palliative Nurses Association, American Association of Nurse Anesthetists, American Association of Critical Care Nurses, and American Academy of Nurse Practitioners were surveyed. Seven hundred and sixty-one questionnaires were analyzed (24.8% response rate). Demographic characteristics of the sample mirrored those of nursing as a whole in the United States. The activities with the highest Mean Activity Index were primarily those involving assessment of pain. Nurses with diploma and Associate Degree preparation rated assessment and nonpharmacologic management higher and communication and collaboration lower than nurses with higher educational preparation. Activities varied somewhat as a reflection of respondents' nursing specialty. The results of the study will be used to refine standards of care, develop nursing education curricula, develop research priorities, and develop a blueprint for a certification examination in pain management nursing. PMID- 11893998 TI - The assessment of discomfort in dementia protocol. AB - Many people with dementia are unable to clearly or consistently verbalize pain symptoms. The Assessment of Discomfort in Dementia (ADD) Protocol is a systematic tool that can be used by nurses to make a differential assessment and treatment plan for both physical pain and affective discomfort experienced by people with dementia. This ADD Protocol is based on the assumption that behaviors associated with dementia are symptoms of unmet physiologic and/or nonphysiologic needs. Steps of the process include a physical assessment, a review of the patient's history, an assessment of affective needs including environmental stress and sensoristasis, and the administration of analgesics. The ADD Protocol is unique in that it uses "as needed" analgesics as a part of the assessment process. The dualistic perspective, in which physical and affective needs are considered in tandem, is an essential and innovative feature of the ADD Protocol. PMID- 11893999 TI - Patients' behavior at the time of injury: effect on nurses' perception of pain level and subsequent treatment. AB - This study examined whether nurses who currently provide direct patient care would perceive and treat pain differently in patients whose pain resulted from activities involving different levels of socially acceptable behavior. Clinical vignettes (differing only with respect to information provided about the patient's behavior at the time of injury) were randomly distributed to all medical/surgical, critical care, and emergency room nurses at this institution. Nurses were asked to record their perception of the patient's pain level and to indicate how much morphine should be given. Nurses (n = 167) did not differ significantly in their mean pain ratings (p =.86) or in the average amount of morphine (p =.80) given to the 2 patients. However, medical/surgical nurses indicated that they would give significantly less morphine to the patient who was presented as engaging in less socially acceptable behavior at the time of injury (p =.03). Overall, significantly more nurses correctly increased the next dose of morphine for the patient who was presented as having injured him or herself while participating in more socially acceptable behavior (p =.003). Thus, although the level of social acceptability of the patient's behavior at the time of injury did not significantly affect the average pain rating or morphine dose that the nurse would have reportedly given, there is evidence that the nurses would have been less aggressive in ensuring adequate pain treatment in the patient exhibiting less socially acceptable behavior. PMID- 11894000 TI - A comparison of the effectiveness of an opioid analgesic and a nonpharmacologic intervention to improve incentive spirometry volumes. AB - The purpose of this study was to compare the effects of opioid analgesics and relaxation as interventions for incentive spirometry in postcardiovascular surgery patients. With the use of incentive spirometry change scores from pre- to posttreatment, 72 patients received either relaxation, opioid analgesia, or no intervention. Pre- to posttreatment differences in achieved spirometry volumes were significantly different for the 3 groups (F[2, 69] = 7.32; p =.002), whereas no difference in spirometry volume was found between the opioid and relaxation groups. Relaxation was as effective as opioid analgesia in permitting postcardiovascular surgical patients to deep breathe. PMID- 11894002 TI - Current and future perspectives in advanced bladder cancer: is there a new standard? AB - The methotrexate/vinblastine/doxorubicin/cisplatin (MVAC) regimen has been the standard treatment in patients with locally advanced and metastatic urothelial cancer for the past 15 years. The minimal or moderate survival benefit-depending on prognostic features-and the severe toxicity associated with the MVAC regimen have made the search for new drugs and drug combinations of utmost importance to increase efficacy and/or decrease toxicity. In this respect, the taxanes and gemcitabine are promising new drugs. Paclitaxel and docetaxel as single agents have yielded overall response rates of 7% to 56%, depending on whether the patients have received prior chemotherapy for metastatic disease. The combination of paclitaxel and cisplatin has been explored in three studies with a total of 104 evaluable patients, a pooled overall response (OR) rate of 61%, and a complete response (CR) rate of 20%. There are two studies of docetaxel and cisplatin with a total of 91 evaluable patients, an OR rate of 54%, and a CR rate of 16%. The OR rate for paclitaxel and carboplatin in six studies was 43%, with a CR rate of 13%; however, the reported median survival was only 8.5 to 9.5 months. The OR rate for single-agent gemcitabine based on five studies was 26%, with a CR rate of 9%, which was apparently independent of whether the patients had received prior chemotherapy. The OR rate for gemcitabine and cisplatin in four phase II studies ranged from 41% to 57%, with a CR rate of 15% to 22% and a median survival of 12.5 to 14.3 months. Based on the encouraging results for the combination of gemcitabine and cisplatin (GC), a randomized phase III trial comparing GC and MVAC was begun in late 1996. This study of 405 randomized patients showed that the two regimens were associated with similar response rates, time to progression, and overall survival, whereas GC was associated with less toxicity than MVAC. On the basis of this superior risk-benefit ratio, the GC regimen should be favored as a new standard treatment in patients with locally advanced and metastatic urothelial cancer. Other promising combinations include gemcitabine and paclitaxel, with or without cisplatin, and the combination of ifosfamide, paclitaxel, and cisplatin. The triple combination of gemcitabine, paclitaxel, and cisplatin has yielded an OR rate of 78%, a CR rate of 28%, and a median survival of 24 months. An international phase III trial comparing this triple combination with GC in patients with locally advanced and metastatic urothelial cancer has now been initiated. PMID- 11894004 TI - Novel gemcitabine-containing triplets in the management of urothelial cancer. AB - Chemotherapy has been the cornerstone of treatment of advanced urothelial cancer. For a decade, the combination regimen of methotrexate/vinblastine/doxorubicin/cisplatin has been considered the standard for these patients. The need for improved efficacy and reduced toxicity of a predominantly palliative therapy has propelled efforts for new drug development. Of the newly identified agents with documented activity, both gemcitabine and paclitaxel have been evaluated with a platinum and have been incorporated into multiagent chemotherapy combinations. Phase II data from two gemcitabine-based triplets are currently available. Combination gemcitabine/paclitaxel/cisplatin and gemcitabine/paclitaxel/carboplatin have high levels of activity with overall and complete response rates of 76% and 26%, respectively, for the former and 68% and 32%, respectively, for the latter combination. The role of gemcitabine-based multiagent combinations compared with standard therapy awaits evaluation in prospectively randomized trials. PMID- 11894003 TI - Gemcitabine doublets in advanced urothelial cancer. AB - Gemcitabine was identified as an active agent in the treatment of urothelial cancer early in its clinical development. A gemcitabine/cisplatin regimen has been shown to lead to comparable survival in a phase III comparison to methotrexate/vinblastine/doxorubicin/cisplatin in the metastatic setting with less toxicity. Nonetheless, cisplatin-related toxicity is not inconsequential. Renal insufficiency limits wide applicability and long-term survival remains poor. A number of additional doublet combinations have thus been investigated. Substitution of carboplatin for cisplatin is feasible but leads to an apparent lower complete response rate. Likewise, combinations of gemcitabine and a taxane are feasible, but with somewhat discouraging response rates. A combination of doxorubicin and gemcitabine has been reported to lead to a 36% complete response rate, but this has not been confirmed. Combinations with targeted therapeutic agents such as the epidermal growth factor receptor inhibitors and trastuzumab have great potential, but the clinical studies have not yet been completed. PMID- 11894005 TI - Gemcitabine-based combination treatment of pancreatic cancer. AB - Since the introduction of gemcitabine (Gemzar; Eli Lilly and Company, Indianapolis, IN), pancreatic cancer may no longer be regarded as a completely chemotherapy-resistant tumor. Good treatment tolerability and a low incidence of side effects are clear advantages of single-agent gemcitabine and enable its integration into combination regimens. Currently, the most widely used regimens involve combination partners such as 5-fluorouracil (5-FU), cisplatin, and docetaxel. Combinations of gemcitabine with cisplatin or 5-FU appear comparably active and tolerable. Comparative analysis of multiple phase II studies performed with gemcitabine/cisplatin showed response rates in the range of 11.4% to 58% and median survival times of 7.4 to 10 months, whereas various gemcitabine/5-FU-based regimens achieved response rates of 3.7% to 25% and median survival times of 4.4 to 10.3 months. In view of the great variety of schedules and inconclusive treatment results, an optimal regimen for the combination of 5-FU and gemcitabine still needs to be defined. Although the combination of gemcitabine with docetaxel has demonstrated activity, data showing a clear survival benefit are not yet available. It is also premature to evaluate the activity of combinations with irinotecan or oxaliplatin. Four-drug regimens indicate a possible improvement in treatment outcome. However, their application may be limited to selected patients with good performance status. PMID- 11894006 TI - Brief communication: use of the multitargeted antifolate pemetrexed (Alimta) in genitourinary cancer. AB - Pemetrexed (Alimta, LY231514) is a novel, multitargeted antifolate that is broadly active in a wide variety of solid tumors, including genitourinary malignancies. This agent has also shown clinically relevant activity in combination with other agents, including gemcitabine (Gemzar; Eli Lilly and Company, Indianapolis, IN). Further investigation is warranted in advanced disease and adjuvant settings. PMID- 11894007 TI - Future directions for gemcitabine in the treatment of genitourinary cancer. AB - Because of an annual morbidity of 225,000 patients and mortality of over 56,000 patients per year in the United States from metastatic genitourinary malignancies, there is a great need for new systemic agents. With its activity and low toxicity, gemcitabine has begun to play a growing role in genitourinary cancer treatment and clinical trials. Substantial activity has been reported for gemcitabine combinations in the treatment of bladder cancer (median survival in one study of nearly 20 months) and for gemcitabine alone or in combinations in testicular cancer patients. Lower (but real) levels of activity have also been observed for gemcitabine combinations in renal carcinoma (17% response rate) and for monotherapy in hormone-refractory prostate cancer (7%). These data suggest the need for further trials of gemcitabine alone or in combinations in genitourinary cancer patients. PMID- 11894008 TI - Seminars in oncology. Introduction. PMID- 11894009 TI - Epidermal growth factor receptor family in lung cancer and premalignancy. AB - Lung cancer, like many other epithelial malignancies, is thought to be the outcome of genetic and epigenetic changes that result in a constellation of phenotypic abnormalities in bronchial epithelium. These include morphologic epithelial dysplasia, angiogenesis, increased proliferative rate, and changes in expression of cell surface proteins, particularly overexpression of epidermal growth factor receptor (EGFR) family proteins. The EFGR family is a group of four structurally similar tyrosine kinases (EGFR, HER2/neu, ErbB-3, and ErbB-4) that dimerize on binding with a number of ligands, including EGF and transforming growth factor alpha. Epidermal growth factor receptor overexpression is pronounced in virtually all squamous carcinomas and is also found in > or = 65% of large cell and adenocarcinomas. It is not expressed in situ by small cell lung carcinoma. Overexpression of EGFR is one of the earliest and most consistent abnormalities in bronchial epithelium of high-risk smokers. It is present at the stage of basal cell hyperplasia and persists through squamous metaplasia, dysplasia, and carcinoma in situ. Recent studies of the effect of inhibitors of receptor tyrosine kinases suggest that patterns of coexpression of multiple members of the EGFR family could be important in determining response. Intermediate endpoints of such trials could include monitoring of phosphorylation levels in signal transduction molecules downstream of the receptor dimers. These trials represent a new targeted approach to lung cancer treatment and chemoprevention that will require greater attention to molecular endpoints than required in past trials. PMID- 11894010 TI - Overview of rationale and clinical trials with signal transduction inhibitors in lung cancer. AB - Most cellular proto-oncogenes encode proteins that participate in signaling pathways by which cells receive and execute instructions that lead to mitogenesis, differentiation, lineage determination, cell migration, extracellular matrix production, and apoptosis, among others. These proto oncogene protein products include transmembrane receptor tyrosine kinases and receptor substrates, serine/threonine kinases, receptor adaptor molecules, low molecular-weight GTPases, and transcription factors that, when overexpressed or mutationally activated, can lead to cell transformation and tumor progression. The large number of oncogenic protein tyrosine kinases plus the rare presence of phosphotyrosine in nontransformed cells argue persuasively that tyrosine phosphorylation and activation of signaling molecules downstream from receptor tyrosine kinases are critical events in growth control and transformation and are, therefore, rational targets for anticancer molecular therapies. We will review some of the more recent treatment strategies in non-small cell and small cell lung cancer targeted to dysregulated signaling pathways that are causally associated with tumor maintenance and progression. PMID- 11894011 TI - Epidermal growth factor receptors as a target for cancer treatment: the emerging role of IMC-C225 in the treatment of lung and head and neck cancers. AB - Epidermal growth factor receptor is one of four receptors critical to cellular proliferation, differentiation, and survival, and is widely expressed in malignant tissue, particularly in squamous cell carcinoma of the head and neck. Expression has been associated with malignant progression, inhibition of apoptosis, neoplastic angiogenesis, enhanced metastatic potential, and both chemoresistance and radioresistance. IMC-C225 is a chimeric monoclonal antibody that targets extracellular epidermal growth factor receptor; it has shown both in vitro and in vivo antitumor activity in tumor cells lines expressing epidermal growth factor receptor, including heightened radiation response in vitro in cultured human squamous cell carcinoma and enhancement of taxane- and platinum induced cytotoxicity in non-small cell lung cancer xenografts. In A431 head and neck squamous cell xenografts, IMC-C225 administered both before and after radiation therapy yields a radiation enhancement factor of 3.62, attributable to both tumor necrosis and antiangiogenesis. In phase I pharmacokinetic studies, IMC C225 has a long half-life, lending itself to convenient weekly administration. It has shown a favorable toxicity profile, limited primarily to allergic and dermatologic reactions, the latter characterized by a self-limited, sterile, acneiform rash. Anaphylaxis is rare. Standard treatment entails a loading dose of 400 mg/m(2) at week 1, followed by a maintenance dose of 250 mg/m(2) weekly. An ongoing phase III international multicenter, randomized study in locally advanced squamous cell carcinoma of the head and neck is evaluating therapeutic radiation therapy, either alone or in conjunction with IMC-C225. In a pilot trial, six of nine patients with platinum-exposed squamous cell carcinoma of the head and neck exhibited objective response. In an ongoing phase II trial in patients with stable or progressive disease on platinum-based therapy, the preliminary response rate is approximately 20%, far higher than one would expect with standard salvage regimens. The Eastern Cooperative Oncology Group has completed a placebo controlled phase III registration trial assessing cisplatin 100 mg/m(2) every 4 weeks with or without IMC-C225. Three separate phase II trials in non-small cell lung cancer have been launched: one trial tests IMC-C225 in combination with standard paclitaxel/carboplatin; another integrates IMC-C225 into the gemcitabine/carboplatin combination in treatment-naive patients; and a third trial evaluates IMC-C225 in combination with docetaxel 75 mg/m(2) every 3 weeks in the second-line setting. PMID- 11894012 TI - ZD1839, a selective epidermal growth factor receptor tyrosine kinase inhibitor, alone and in combination with radiation and chemotherapy as a new therapeutic strategy in non-small cell lung cancer. AB - The epidermal growth factor receptor is overexpressed in a majority of non-small cell lung cancers and has been associated with a poor prognosis. Preclinical studies have shown that ZD1839, an oral anilinoquinazoline, targets the epidermal growth factor receptor-associated tyrosine kinase, reversibly inhibiting critical downstream signaling and resulting in cancer cell growth arrest. Potent antitumor effects have been observed in human lung tumor xenograft models. Preclinical studies have shown additive to synergistic effects when ZD1839 is combined with radiation or chemotherapy in colon, head and neck, and non-small cell lung cancers. Phase I clinical trials have shown modest dose-related toxicity, and antitumor activity has been reported in a variety of malignancies including lung cancer. Future studies will certainly combine ZD1839 with chemotherapy or radiation. ZD1839 also may be effective as a chemoprevention agent because premalignant lesions often overexpress epidermal growth factor receptor. PMID- 11894013 TI - Therapeutic potential of ABX-EGF: a fully human anti-epidermal growth factor receptor monoclonal antibody for cancer treatment. AB - Overexpression of the epidermal growth factor receptor (EGFR) has been observed in a wide variety of human cancers and is associated with a poor clinical prognosis. In many cases, growth of the tumor cells is dependent on EGFR-mediated signals, because inhibition of binding of factors to the EGFR leads to cell death. Using XenoMouse technology, a fully human EGFR-specific monoclonal antibody, ABX-EGF, with high affinity (5 x 10(-11) mol/L) has been generated. ABX EGF blocks binding of both epidermal growth factor and transforming growth factor alpha to the EGFR, inhibits tyrosine phosphorylation of the EGFR, and inhibits cellular proliferation. In vivo, ABX-EGF not only blocks formation of human epidermoid carcinoma A431 xenografts in athymic mice, but also mediates therapeutic elimination of established tumors and acts cooperatively with chemotherapeutics in mediating tumor regression. These observations provide a strong basis for the development of ABX-EGF as a therapeutic agent for human solid tumors that overexpress EGFR. PMID- 11894014 TI - HER2/neu expression in malignant lung tumors. AB - Despite intensive treatment efforts, the prognosis for lung cancer is very poor; less than 15% of patients survive 5 years. Trastuzumab, a monoclonal antibody targeting the HER2/neu protein receptor, is effective in the treatment of metastatic breast cancer and may be useful in the treatment of non-small cell lung cancer (NSCLC). Using the HercepTest (Dako; Carpenteria, CA), 25% of NSCLC show 2+ or greater HER2/neu expression, but only 6% to 8% of NSCLC tumors have 3+ overexpression. Positive HER2/neu expression is most often seen in adenocarcinomas compared with squamous cell carcinomas or large cell carcinomas, and is rarely seen in small cell lung cancer. As determined by fluorescence in situ hybridization analysis, the high degree of HER2/neu gene expression and gene amplification seen in breast cancer is lower in NSCLC. Polysomy is the cause of increased HER2/neu expression in most NSCLC. Prospective clinical studies with trastuzumab in lung cancer are ongoing. Future studies in NSCLC need to include immunohistochemistry and fluorescence in situ hybridization analysis to determine the method of choice for evaluating clinically relevant HER2/neu-positive tumors. PMID- 11894015 TI - Trastuzumab in the treatment of non-small cell lung cancer. AB - Trastuzumab is a humanized monoclonal antibody that binds to human epidermal growth factor-2 (HER2) and is approved by the US Food and Drug Administration for the treatment of advanced breast cancer that overexpresses HER2/neu protein. Preclinical data suggests a role for trastuzumab in the treatment of non-small cell lung cancer (NSCLC). HER2 protein is overexpressed in 20% to 66% of resected NSCLC tumors, and has been shown to predict poor patient outcome in multiple series. Experiments with NSCLC cell lines show that HER2 overexpression increases chemoresistance, invasiveness, and metastatic potential of the cells. In mouse xenograft experiments, trastuzumab halts tumor growth and is synergistic with cytotoxic chemotherapy. Ongoing phase II trials are showing that trastuzumab can be added to standard chemotherapy in the treatment of patients with advanced NSCLC without additional toxicity, and with promising efficacy. Whether trastuzumab will show a clear benefit for patients with NSCLC, either alone or in combination with established chemotherapy, remains to be proven in phase III testing. PMID- 11894016 TI - Angiogenesis inhibitors in clinical development for lung cancer. AB - The use of tumor angiogenesis as a therapeutic target is based on extensive literature showing the dependence of tumors on the process of angiogenesis for growth, invasion, and metastasis. Seminal work performed by Folkman three decades ago determined that tumors beyond the size of approximately 2 mm require angiogenesis for subsequent growth and development. This basic hypothesis stimulated research in the field of angiogenesis and has resulted in the identification of factors that both enhance and inhibit this "angiogenic switch." The intent of this article is to present data on several angiogenesis inhibitors that are currently undergoing clinical evaluation in cancer patients. These agents may be particularly useful in the treatment of lung cancer, both as adjunctive therapy in early-stage or locally advanced disease, as well as in combination strategies with platinum-based therapy in metastatic disease. Although angiogenesis inhibitors have been in clinical trials for the past decade, there has been a shift in recent years towards the development of more mechanism-based and receptor-targeted agents. Interestingly, no antiangiogenic agent has been approved as such for use in cancer, perhaps because of the challenges involved in the clinical development of these novel agents. These include the potential requirement for long-term administration, difficulties in deriving biologically efficacious doses in early clinical trials, and the inability to use tumor regression as a primary endpoint in phase II trials. PMID- 11894017 TI - Matrix metalloproteinases and matrix metalloproteinase inhibitors in lung cancer. AB - Preclinical studies have provided evidence that matrix metalloproteinases (MMPs), a family of zinc-containing proteolytic enzymes, facilitate tumor invasion, the establishment of metastases, and the promotion of tumor-related angiogenesis. Matrix metalloproteinase inhibitors (MMPIs) have been shown to inhibit tumor growth and dissemination in preclinical models. Not all lung cancers express the MMPs believed to be most important in promoting the neoplastic process, and there are conflicting reports regarding the prognostic significance of MMPs in lung cancer. However, it is possible that these observations are because of limitations in the procedures for measuring MMPs. Many investigators believe that MMPs are universally involved in tumor progression; this hypothesis was the basis for initiating seven phase III MMPI trials in lung cancer. Four studies were closed at completion of the predefined accrual goal, and three were closed early. There were no significant differences in survival in a non-small cell lung cancer prinomastat study, and in a small cell lung cancer marimastat trial. The results of the remaining five studies have not been reported. At this point it appears that MMPIs will probably not play a major role in the treatment of advanced lung cancer patients. PMID- 11894018 TI - Preclinical and clinical studies of docetaxel and exisulind in the treatment of human lung cancer. AB - Lung cancer is the leading cause of cancer death in the United States. The majority of patients with non-small cell lung cancers present with inoperable disease because of the presence of metastases to regional lymph nodes or other metastatic sites. About one third of patients have stage IV disease with metastases to distant organs at the time of diagnosis. The prognosis for these patients is very poor. With best supportive care the median survival is only 4 months and the 1-year survival rate is 10% to 15%. Current chemotherapy combinations improve the survival and quality of life for patients with advanced non-small cell lung cancer. With two-drug combinations, median survival is increased to 8 months or more and 1-year survival is increased to 35% to 40%. Still, complete response rates are low and more than 80% of patients die within 1 year of diagnosis. The improvements created by current therapies led to studies of chemotherapy in the second-line setting. Docetaxel has been shown to improve survival of patients who failed platinum-based chemotherapy and was approved by the U.S. Food and Drug Administration for therapy in this setting. However, response rates were very low and survival very short. Therefore, new therapies are urgently needed. Exisulind is a novel oral anticancer agent that holds promise for the treatment of patients with advanced non-small cell lung cancer. Exisulind was originally developed as a chemoprevention agent for colorectal cancer. Preclinical studies showed that exisulind could prevent polyp formation and inhibit the growth of colorectal cancers. Subsequent preclinical studies showed that exisulind also inhibited the growth of human breast, prostate, and lung cancers. Phase I clinical studies showed that twice-daily oral doses could be given safely and would provide peak concentrations that were equivalent to those required for in vitro effects. These observations lead to the studies of the combination of exisulind and docetaxel in preclinical and clinical studies in human lung cancer described in this article. PMID- 11894019 TI - Genetic and immunologic therapies for lung cancer. AB - The poor overall survival of lung cancer patients treated with conventional therapies (chemotherapy, radiation therapy, and surgery) mandate novel approaches to treatment. Two novel approaches to treat lung cancer include gene therapy and immunologic therapy. Both treatments have preclinical data suggesting potential clinical use. In gene therapy, the identification of specific genes critical to the development of carcinogenesis has offered the opportunity to target these genes or their products for treatment. One possible gene therapy strategy that has been pursued in phase I and II lung cancer trials is to replace nonfunctional tumor suppressor genes such as mutated or deleted p53 genes with wild-type p53 genes by adenoviral gene transfer (Ad-p53). Transduction of the tumors has been accomplished with direct intratumoral injection or broncheoalveolar lavage. These studies have identified a potential role for radiosensitization of previously radiation-resistant local tumors by combining Ad-p53 with radiation or possibly chemoradiation. Another novel strategy that may allow systemic treatment of lung cancers is immunologic therapies. Immunotherapies have focused on augmenting the immune response to cancer by passive strategies (e.g., antivascular endothelial growth factor) or active nonspecific (e.g., interferon), or by specific (e.g., anti-idiotype therapy) strategies. These novel strategies are currently in clinical trials and will potentially allow additional therapeutic options for patients resistant to conventional therapies. PMID- 11894021 TI - Determinants of response and resistance to cytotoxics. AB - There is an underlying feeling in the oncology community that chemotherapy has reached a therapeutic "glass ceiling" in non-small cell lung cancer, and that we will never attain the acceptable survival rates that are just beyond our reach. Multiple clinical trials have tested doublets, triplets, and sequential chemotherapy in what is often regarded as a futile attempt to break through this ceiling. Recent large randomized trials have not shown that any of these combinations is superior to any of the others. Nevertheless, the analysis of DNA and RNA from patients' serum can permit us to assess genes that may be specific targets of certain cytotoxic agents and others that may be markers of multidrug resistance. With this information, we will hopefully be able to create guidelines for individualized chemotherapy. With this in mind, the Spanish Lung Cancer Group has designed a trial to test the involvement of various genes in resistance to gemcitabine and cisplatin. With the gene corral that will emerge from this trial, we will create an up-front diagnostic test for selecting the most appropriate gemcitabine plus cisplatin regimen for the treatment of non-small cell lung cancer. PMID- 11894022 TI - New agents, new treatment, and antiemetic therapy. AB - With the introduction of newer antineoplastic agents, the challenge for supportive care is enlarging. For the most part, these targeted therapies are given in addition to more classic anticancer drugs, either in combination or in sequence, underscoring the need for prevention of emesis and attention to all treatment toxicities. Strategies for preventing these side effects need to be investigated while appropriately addressing more familiar problems such as emesis. Two directions for improving emetic control are clear. First is the development of newer antiemetics that enhance current control rates, and second is the proper use of existing agents in all emetic settings. This report outlines the rationale behind the development of neurokinin type 1 antagonists, and reviews consensus recommendations in the prevention of acute and delayed emesis. PMID- 11894023 TI - Meaningful survival in lung cancer patients. AB - Lung cancer is diagnosed at an advanced stage in a majority of patients. Recent advances made in the treatment of lung cancer have resulted in a prolongation in survival and an improvement in quality of life. The majority of lung cancer patients experience multiple symptoms, which result from both the cancer and its treatment. Fatigue and anemia cause significant morbidity and impaired quality of life among patients with lung cancer. They also contribute to a suboptimal response to treatment modalities such as radiation, decreased performance status, and poor patient compliance with treatment. The impact of anemia is frequently under-recognized. Improvements in survival made with multimodality therapy can only be meaningful when combined with interventions to improve symptoms and overall quality of life. Prompt recognition of these problems and early intervention are an integral part of cancer therapy. In this review, we discuss the impact of anemia and the treatment options that could contribute to a truly meaningful survival in lung cancer patients. PMID- 11894020 TI - Tirapazamine: prototype for a novel class of therapeutic agents targeting tumor hypoxia. AB - Preclinical models in vitro and in vivo have shown that tumor hypoxia alters the malignant cell phenotype, selecting for p53 mutations, stimulating angiogenesis and metastasis, and markedly reducing the efficacy of both radiotherapy and chemotherapy. Similarly, clinical studies measuring pretreatment tumor oxygen status confirm that the presence of hypoxia confers a negative impact on local control, disease-free survival, and overall survival. Despite these data and extensive past research efforts, the promise of developing selective hypoxic-cell sensitizers has been largely unfulfilled. In contrast, tirapazamine is the rationally designed prototype for a new class of therapeutic agents targeting tumor hypoxia: hypoxic cytotoxins. Tirapazamine is bioreductively activated in hypoxic cells and has been shown to potentiate the cytotoxicity of radiation and a number of chemotherapeutic drug classes, in particular platinum compounds and taxanes. This article reviews the preclinical and clinical development of tirapazamine, as well as current trials in non-small cell lung cancer designed to provide proof of principle for this new category of cancer therapeutics. PMID- 11894024 TI - Bisphosphonates in the treatment of bone metastases. AB - Bone metastases are one of the most common problematic complications of advanced cancers. In addition to causing significant pain, bone metastases often result in fractures and debilitation. Stimulation of osteoclast activity by factors secreted by tumor cells is believed be the primary mechanism of bone destruction. Bisphosphonates inhibit osteoclast-related bone resorption, and have become standard therapy in the treatment of hypercalcemia of malignancy and postmenopausal osteoporosis. More recently, bisphosphonates have been shown to decrease pain and skeletal fractures associated with bone metastases. Structural changes in bisphosphonates influence their relative potency as well as other potentially beneficial effects such as inhibition of tumor growth factors, alteration of adhesion molecules, and apoptosis of tumor cells. PMID- 11894026 TI - Bacterial translocation or lymphatic drainage of toxic products from the gut: what is important in human beings? PMID- 11894027 TI - The important role of operations in the management of anaplastic thyroid carcinoma. AB - BACKGROUND: Anaplastic thyroid carcinoma is a rare and highly lethal neoplasm. We investigated whether operations have an impact on the survival of patients who have anaplastic carcinoma without distant metastasis. METHODS: Between 1989 and 1999, 40 consecutive patients with anaplastic carcinoma, without distant metastasis at the time of presentation or during local treatment, were reviewed. The cumulative survival rates and 1-year survival rates were compared. RESULTS: Eleven patients had a small focus of anaplastic carcinoma in a differentiated carcinoma, and 29 patients had ordinary anaplastic thyroid carcinoma. Surgical debulking was performed in 26 patients. Radiotherapy was used for 31 patients and chemotherapy for 19 patients. The 1-year survival rates of the patients with incidental anaplastic carcinoma, ordinary anaplastic carcinoma who underwent operations, and ordinary anaplastic carcinoma who did not undergo operations were 73%, 60%, and 21%, respectively. A significantly higher cumulative survival rate was observed in patients with incidental anaplastic carcinoma than in those with ordinary anaplastic carcinoma. A significantly better outcome was obtained by surgical debulking of ordinary anaplastic carcinoma. CONCLUSIONS: Patients with incidental anaplastic carcinoma tended to have a good outcome, but some had a poor prognosis. Surgical debulking improved the outcome of patients with ordinary anaplastic carcinoma. PMID- 11894028 TI - Distribution of lymph node micrometastasis in pN0 well-differentiated thyroid carcinoma. AB - BACKGROUND: The role of lymph node dissection in the treatment of differentiated thyroid carcinoma remains controversial, and the benefit of therapy is debatable. This study was designed to identify the precise localization of lymph node micrometastases (LNMM) and map their cervical involvement in relation with the tumor location within the thyroid gland. METHODS: A total of 2551 cervical lymph nodes were obtained from 80 patients with well-differentiated thyroid cancer. They were diagnosed as clear lymph nodes by hematoxylin and eosin stain and then examined immunohistochemically with cytokeratins (AE1/AE3) for evidence of micrometastases. RESULTS: Forty-two patients out of 80 (53%) had LNMM. Forty eight patients (60%) had the tumor confined to only one third of 1 of the 2 lobes of the thyroid gland or isthmus. The frequencies and locations of LNMM in patients were 50% (3/6) in the deep upper cervical nodes, with tumors localized in the upper third; 31% (5/16) in the paraglandular nodes, with tumors affecting the middle third; 63% (12/19) in the paratracheal nodes, with tumors affecting the lower third of the thyroid lobe; and 71% (5/7) in the pretracheal nodes in the isthmus-located tumor. All the LNMM occurred on the ipsilateral side of the tumor. CONCLUSIONS: When thyroid carcinoma is located in the upper third of the thyroid lobe, the LNMM are found in the direction of upward lymphatic flow. When the tumor is located in the lower third or isthmus, LNMM are directed downward. In addition, early thyroid carcinoma micrometastases do not cross the midline but remain on the ipsilateral side of the tumor. PMID- 11894029 TI - Parathyroidectomy in familial hypercalcemia with clinical characteristics of primary hyperparathyroidism and familial hypocalciuric hypercalcemia. AB - BACKGROUND: Familial primary hyperparathyroidism is associated with tumor susceptibility syndromes, which are unrelated to mutations in the calcium receptor gene. This study describes parathyroidectomy in a kindred with hypercalcemia due to a heterozygous point mutation in the calcium receptor gene. METHODS: Seventeen family members were studied, and postoperative follow-up averaged 5.1 years. RESULTS: Radical parathyroid resection with total parathyroid remnants of 10 to 20 mg or total parathyroidectomy with autotransplantation normalized the serum calcium and parathyroid hormone values in 12 family members. Persistent hypercalcemia was noted in 3 of 5 patients subjected to less radical procedures. Diffuse to nodular hyperplasia and microscopic findings, interpreted incorrectly as a single adenoma, were found. Weight of the parathyroid tissue increased with the age of the patients (P <.05), and almost one third of them (29%) had 1 to 3 atypically located glands. There were no patients with recurrent hypercalcemia during follow-up. CONCLUSIONS: The heterozygous inactivating mutation of the calcium receptor gene of this family is accompanied by mild increases in parathyroid gland x weight and diffuse parathyroid hyperplasia with possibly secondary genetic events causing nodule formation. Radical parathyroid resection is advocated in this hypercalcemic disorder, which may represent an intermediary stage between primary hyperparathyroidism and familial hypocalciuric hypercalcemia. PMID- 11894030 TI - Screening for primary hyperparathyroidism before thyroid surgery: A prospective study. AB - BACKGROUND: Misdiagnosed primary hyperparathyroidism (PHPT) during thyroid surgery may lead to a difficult reoperation. Because PHPT is often asymptomatic, calcium measurements have been recommended before thyroid surgery, but no study has focused on the results of a prospective PHPT screening. METHODS: The prospective study of 748 patients consisted of 2-step screening of calcium measurement in all patients (normal range, 2.2 to 2.6 mmol/L, 8.8 to 10.4 mg/dL). If the calcium level was greater than 2.49 mmol/L (9.9 mg/dL), parathyroid hormone level (PTH; normal range, 11 to 65 pg/mL) and second calcium measurements were obtained. Positive screening was defined by 2 calcium levels greater than 2.49 mmol/L (9.9 mg/dL) and PTH level greater than 49 pg/mL. In patients with negative screening, we evaluated the number of parathyroid incidentalomas. In patients with positive screening, we rated parathyroid adenomas discovered as "easily accessible" or "requiring specific dissection." We assumed that the former could have been incidentally found by a surgeon unaware of calcium value. The cost estimation was based on French national health system databases. RESULTS: In the 9 patients with positive screening, 9 had parathyroid adenomas, 3 of them requiring specific dissection. In the 739 patients with negative screening, 12 had surgical incidentalomas and 2 had postoperative PHPT diagnosis. CONCLUSIONS: Our screening was not exhaustive, but it avoided a reoperation for missed PHPT in 3 patients. Population screening cost less than 3 reoperations. Other strategies, more exhaustive and/or cost-effective, should be investigated. PMID- 11894031 TI - Closure of lacerations and incisions with octylcyanoacrylate: a multicenter randomized controlled trial. AB - BACKGROUND: Most lacerations and surgical incisions are closed with sutures or staples. Octylcyanoacrylate tissue adhesive (OCA) was recently approved for use in the United States. We compared the cosmetic appearance of lacerations and incisions repaired with OCA versus standard wound closure methods (SWC). METHODS: A multicenter randomized clinical trial including patients with simple lacerations or surgical incisions was conducted at 10 clinical sites. Patients were randomly assigned to treatment with OCA or SWC. Follow-up was performed at 1 week and at 3 months to determine infection rates and cosmetic outcome. RESULTS: Eight hundred fourteen patients with 924 wounds (383 traumatic lacerations, 235 excisions of skin lesions or scar revisions, 208 minimally invasive surgeries, and 98 general surgical procedures) were enrolled. Groups were similar in baseline characteristics. Wound closure with OCA was faster than with SWC (2.9 vs 5.2 minutes, P <.001). At 1 week infection rates were similar (OCA, 2.1% vs SWC, 0.7%; P =.09) and fewer OCA wounds were erythematous (18% vs 36%, P <.001). There were no differences in wound dehiscence rates (OCA, 1.6% vs SWC, 0.9%; P =.35). At 3 months there was no difference in the percent of wounds with optimal appearance (OCA, 82% vs SWC, 83%; P =.67). CONCLUSIONS: Repair of traumatic lacerations and surgical incisions with OCA is faster than with SWC, and cosmetic outcome is similar at 3 months. PMID- 11894032 TI - Comparison between periareolar and peritumoral injection of radiotracer for sentinel lymph node biopsy in patients with breast cancer. AB - BACKGROUND: The technique of sentinel lymph node (SLN) biopsy in patients with breast cancer varies among reports, and the optimal method remains to be established, particularly with regard to the site of radiotracer injection. The aim of this study was to compare periareolar and peritumoral injection of radiotracer in detecting SLN in patients with breast cancer. METHODS: Patients with T1-2 breast cancer (n = 155) were enrolled in this study. In phase 1 (n = 62), SLN biopsy was performed by using peritumoral injection of blue dye alone followed by backup axillary lymph node dissection. In phase 2, SLN biopsy was performed by using peritumoral injection of blue dye and peritumoral (group A, n = 41) or periareolar (group B, n = 52) injection of technetium 99m tin colloid. RESULTS: In phase 1, the detection rate of SLN was 81% and the false-negative rate was 5.6%, indicating our skill in SLN biopsy. In phase 2, the success rate of lymphoscintigraphy was significantly (P <.001) higher in group B (90%) than in group A (51%). The mean ex vivo radioactivity of SLN in group B (117 counts per second; range, 5 to 900) was also significantly (P <.05) higher than in group A (51 counts per second; range, 8 to 260). In addition, the detection rate of SLN was significantly (P <.05) higher in group B (100%) than in group A (90%). CONCLUSIONS: Periareolar injection of radiotracer for SLN biopsy is superior to peritumoral injection because of its simplicity, achieving a high success rate in lymphoscintigraphy and SLN detection. PMID- 11894033 TI - Early versus delayed carotid endarterectomy after a nondisabling ischemic stroke: a prospective randomized study. AB - BACKGROUND: Although many retrospective and a few prospective studies have analyzed the outcome of early and delayed carotid endarterectomy (CEA) after a recent minor or nondisabling stroke (ie, a minimal and stabilized focal neurologic deficit of acute onset persisting for more than 24 hours and not leading to a handicap or to a significant impairment of daily living activities), the optimal timing of surgery remains uncertain. The purpose of this study was to prospectively compare the perioperative death and stroke rates of CEA performed within 30 days (early group) or more than 30 days (delayed group) after a nondisabling ischemic stroke in patients with carotid bifurcation disease. METHODS: During a 4-year period, of 86 patients experiencing a minor stroke, 45 were randomized to undergo early CEA and 41 to undergo delayed CEA. All patients underwent preoperative cerebral computed tomography, duplex ultrasonographic scanning and angiography of the supra-aortic trunks. All CEAs were carotid eversion endarterectomies and were performed by the same surgeon, using deep general anesthesia, with continuous electroencephalographic monitoring for the selective shunting. The perioperative death and stroke rates were compared between the 2 groups. RESULTS: No perioperative deaths occurred in either group. No recurrent strokes occurred during the waiting period in the delayed group. The incidence of perioperative stroke was comparable in the 2 groups (1 of 45, 2% vs 1 of 41, 2%). The mean follow-up was 23 months (range, 6 to 50 months). Survival rates after 1, 2, and 3 years were similar in the 2 groups. CONCLUSIONS: Early CEA after a nondisabling ischemic stroke can be performed safely with perioperative mortality and stroke rates comparable with those of delayed CEA. The timing of surgery does not seem to influence the benefit of the CEA. PMID- 11894034 TI - Preoperative selective portal vein embolization before hepatectomy for liver metastases: long-term results and impact on survival. AB - BACKGROUND: Some patients cannot undergo curative surgical procedures for liver metastases because of the risk of severe postoperative hepatic failure, which stems from a too-small future remaining liver (FRL). Preoperative portal vein embolization (PVE) is an effective means of creating hypertrophy of the FRL, thus permitting safe hepatic resection. The aim of this retrospective study was to investigate the long-term results of this technique. METHODS: Sixty-eight patients underwent PVE. Of those, 60 (88%) subsequently underwent hepatic resection. Indication for PVE was an estimated FRL ratio (assessed by volumetric computed tomography) of less than 30%. However, if the patient had undergone multiple courses of chemotherapy, the threshold was 40%. The origin of the primary neoplasm was colorectal in 41 patients (68%); in the remaining 19 (32%), the primary neoplasms originated at other sites. RESULTS: Mean growth of the estimated FRL measured by computed tomography 1 month after PVE was 13%. Major complications after hepatectomy occurred in 27% of the patients, and the operative mortality rate was 3%. For the 60 patients who underwent PVE followed by hepatic resection, the 5-year overall survival rate and the disease-free survival rate were 34% and 24%, respectively. The 5-year overall survival rate and the disease-free survival rate of patients with colorectal metastases only were 37% and 21%, respectively. CONCLUSIONS: The long-term survival rate after PVE followed by resection is comparable with the survival rate obtained after resection without preoperative PVE. The 5-year survival rate of patients undergoing PVE followed by hepatectomy justifies the use of this technique. This technique thus increases the suitability of resection as a treatment choice for patients with liver metastases. PVE should number among the therapeutic options available to every hepatic surgeon. PMID- 11894035 TI - Surgical resection combined with chemotherapy for advanced hepatocellular carcinoma with tumor thrombus: report of 19 cases. AB - BACKGROUND: Prognosis of hepatocellular carcinoma (HCC) with tumor thrombus in the main portal vein (MPV), inferior vena cava (IVC), or extrahepatic bile duct (EBD) treated by conventional therapies has been considered poor. This study aimed to evaluate the efficacy of hepatic arterial infusion chemotherapy after surgical resection as an adjuvant therapy or as a treatment for intrahepatic recurrence of HCC with tumor thrombus in MPV, IVC, or EBD. METHODS: Nineteen patients with HCC and tumor thrombus in the MPV, IVC, or EBD who underwent hepatectomy with thrombectomy were reviewed retrospectively. RESULTS: The overall 3-year survival rate was 48.5%. Two patients with postoperative residual tumor thrombus died within 6 months owing to rapid progression of the residual tumor thrombus. Five patients survived more than 5 years after their operations. Tumors disappeared completely in 3 patients after hepatic arterial infusion chemotherapy with a combination of cisplatinum and 5-fluorouracil, and the longest survival period was 17 years and 11 months in a patient with EBD thrombus. CONCLUSIONS: If hepatic reserve is satisfactory, an aggressive surgical approach combined with chemotherapy seems to be of benefit for patients having HCC with tumor thrombus in the MPV, IVC, or EBD. PMID- 11894036 TI - Extent of liver resection influences the outcome in patients with cirrhosis and small hepatocellular carcinoma. AB - BACKGROUND: The long-term outcome after resection of hepatocellular carcinoma (HCC) is influenced by parameters related to the tumor and the underlying liver disease. However, the extent of the resection, which can be limited or anatomical (including the tumor and its portal territory), is controversial. METHODS: Among 64 Child-Pugh A patients with cirrhosis who underwent curative liver resection for small HCC (< or = 4 cm) between 1990 and 1996, 34 patients underwent limited resection with a margin width of at least 1 cm, and 30 patients underwent anatomic resection of at least 1 liver segment with complete removal of the portal area containing the tumor. The 2 groups were comparable in terms of epidemiologic and pathologic parameters. The major end points were: (1) in hospital mortality and morbidity; (2) overall and disease-free survival; and (3) rate and topography of recurrence. RESULTS: The 30-day mortality (6% vs 7%) and morbidity (52% vs 47%) rates after limited and anatomic liver resection were not statistically different. The 5- and 8-year overall survival rates after limited versus anatomic resection were, respectively, 35% versus 54% (P <.05) and 6% versus 45% (P <.05). The 5- and 8-year disease-free survival rates were, respectively, 26% versus 45% and 0% versus 21% (P <.05). Local recurrence was more frequently observed after limited resections than after anatomic resections (50% vs 10%, P <.05). CONCLUSIONS: In patients with cirrhosis and a small HCC, anatomic resection achieves better disease-free survival than limited resection without increasing the postoperative risk. Therefore, anatomical resection should be the treatment of choice and considered as the reference surgical treatment compared with other treatments. PMID- 11894037 TI - Relationship between skill and outcome in the laboratory-based model. AB - BACKGROUND: Recent attention has been directed at developing quantitative assessments of surgical skill. This study aims to demonstrate whether objectively measuring differences in manual dexterity has an impact on a simulated surgical procedure. METHODS: Six general surgical trainees performed 5 polytetrafluoroethylene graft to artery anastomoses on a vascular model by using a standardized technique. Manual dexterity was objectively measured with (1) electromagnetic motion analysis: trackers applied to the backs of hands recorded and analyzed both hand movements and procedural time and (2) 4-parameter evaluation of the final product. Outcome parameters were assessed by (1) rate of anastomotic leakage and (2) smallest cross-sectional area of the anastomosis. RESULTS: The 2 objective measures of manual dexterity correlated closely (Pearson coefficient, 0.423; P <.02). Trainees with better manual dexterity scores produced better outcome measures. Those with better motion analysis scores produced anastomoses that leaked less (Pearson coefficient, 0.514; P <.01) and those with higher global evaluation scores had a larger anastomotic cross sectional area (Pearson coefficient, 0.495; P <.01). Time taken for the procedure did not appear to influence either outcome measure. CONCLUSIONS: There is a significant correlation between objective measures of manual dexterity and the outcome measures in this model. This suggests that the outcome of a procedure can be predicted by measuring surgical skill. PMID- 11894038 TI - Impaired balance of type I and type III procollagen mRNA in cultured fibroblasts of patients with incisional hernia. AB - BACKGROUND: Recent findings of an impaired protein ratio of type I to type III procollagen showed a disturbed collagen metabolism in incisional hernia development. We analyzed the type I and type III procollagen messenger RNA to investigate whether these findings represent the altered extracellular matrix or a primary defect at the transcriptional level. METHODS: We examined cultured skin fibroblasts of patients with incisional or recurrent incisional hernia in comparison with those without any previous incision (control) and those with a skin scar without clinical appearance of a hernia (scar). Immunohistochemical detection of a lowered protein ratio of type I and type III collagen in the hernia skin tissue leads to mRNA expression analysis. The procollagen mRNA and the ratio of type I to type III procollagen mRNA are detected by reverse transcriptase-polymerase chain reaction and Northern blot analysis, the collagens type I and III by Western blot analysis. RESULTS: Reverse transcriptase polymerase chain reaction revealed an increase of type I procollagen mRNA in the incisional and recurrent hernia (0.90 +/- 0.04 and 1.19 +/- 0.04, respectively) compared with stable scar (0.54 +/- 0.02) or healthy tissue (0.43 +/- 0.01). The obvious rise of type III procollagen mRNA to 4.13 +/- 0.04 for incisional, 6.02 +/- 0.03 for recurrent hernia, 2.29 +/- 0.04 for stable scar, and 1.72 +/- 0.03 for the healthy tissue showed a significantly decreased ratio of type I to type III procollagen mRNA in the hernia patients as compared with the controls (P <.01). By Western blot analysis, an increase of type I and type III collagen protein and a significant rise in the corresponding ratio in the recurrent hernia group were detected. CONCLUSIONS: The altered synthesis of type I and type III collagen in cultured skin fibroblasts suggests a disorder of collagen metabolism, at least in patients with recurrent hernia. Hence, a basically impaired wound healing process is likely to contribute to the unsatisfactory results of incisional hernia repair. PMID- 11894039 TI - Granulation tissue regression induced by musculocutaneous advancement flap coverage. AB - BACKGROUND: Clinical experience suggests that granulation tissue may be inhibited by coverage with a musculocutaneous flap. We hypothesized that coverage of an open wound with a musculocutaneous flap would result in regression and apoptosis of the wound's granulation tissue. METHODS: In the first experiment, 32 rats underwent excisional wounding; 16 underwent musculocutaneous flap coverage of their granulation tissue on postwounding day 8, and then 16 rats (8 controls + 8 flaps) were killed on both postwounding days 10 and 12 (2 and 4 days after the flap procedure, respectively). In the second experiment, 18 rats were wounded, and on postwounding day 5 the rats underwent flap coverage (n = 6), wound edge release/mobilization (the first step of the flap procedure) without flap coverage (n = 6), or dressing change only (n = 6); all rats were killed on postwounding day 6 (24 hours after the secondary intervention). Apoptosis was quantified with the terminal deoxynucleotidyl transferase-mediated nick-end labeling assay. RESULTS: Placement of a musculocutaneous flap over an 8-day-old excisional wound in the first experiment increased the apoptotic rate in the granulation tissue from 0% to 1% (controls) to 5% to 10% at both 2 and 4 days after flap coverage (P <.05). Cell population density decreased 50% in the flap-covered granulation tissue compared with the controls (P <.05). In the second experiment, circumferential release of the granulation tissue resulted in an equivalent increase in granulation tissue apoptosis over controls compared to that induced by the full flap procedure. CONCLUSIONS: Coverage of established granulation tissue with a musculocutaneous flap resulted in histologic regression of the wound's granulation tissue after 2 to 4 days of flap coverage and induced at least a 5-fold increase in the apoptotic rate of the granulation tissue. Releasing the wound edge increased granulation tissue apoptosis to a level equivalent to that produced by the musculocutaneous flap procedure, suggesting that alteration of the wound's mechanical environment is responsible for the acute induction of apoptosis in this model. PMID- 11894040 TI - Premenopausal women deposit more collagen than men during healing of an experimental wound. AB - BACKGROUND: From a post hoc analysis a hypothesis was generated that women deposit more collagen in a surrogate test wound than men. The purpose of this study has been to verify this hypothesis prospectively in a controlled study. METHODS: Post hoc analyses were done on 37 volunteers (study A). The prospective trial included 47 smoking volunteers (study B). Outcome measures were deposition levels of collagen (hydroxyproline) and protein during a period of 10 days in subcutaneously implanted tubes of expanded polytetrafluoroethylene. RESULTS: The mean increments of collagen deposition levels in women as compared with men were 56% (P <.01) in study A and 74% (P <.001) in study B. The mean increase in the ratio collagen/total protein was 74% (P <.001) and 69% (P <.001), indicating that the increase was specific for collagen. CONCLUSIONS: The studies show that deposition in a miniature subcutaneous test wound of collagen, but not noncollagenous protein, is promoted in women as compared to men. These findings may relate to the observation in some reports indicating higher rates of compromised postoperative wound healing in men. PMID- 11894041 TI - P38 mitogen-activated protein kinase inhibition attenuates ischemia-reperfusion injury of the rat liver. AB - BACKGROUND: Several studies have implicated the mitogen-activated protein kinase (MAPK) signal pathway in non-hepatic organ ischemia-reperfusion injury. However, the role of p38 MAPK in hepatic ischemia-reperfusion injury remains unclear. This study investigated the role of p38 MAPK in hepatic ischemia-reperfusion injury. METHODS: Male Sprague-Dawley rats were divided into 4 groups (sham, FR-only, control, and FR-treated groups). The animals in the control and FR-treated groups were subjected to 30 minutes of warm ischemia with congestion of the gut. The FR only and FR-treated groups received FR167653 (FR), which is a novel p38 MAPK inhibitor. The serum levels of aspartate transaminase, alanine transaminase, lactate dehydrogenase, tumor necrosis factor-alpha (TNF-alpha), and interleukin 1beta (IL-1beta) were measured (each, n = 6). Liver tissue blood flow was measured at pre-ischemia, end-ischemia, and 30, 60, 90, and 120 minutes after reperfusion (each, n = 4). The liver tissues in the control and FR-treated groups were excised for p38 MAPK and c-Jun N-terminal kinase (JNK) analyses and histopathology (each, n = 4). RESULTS: Serum levels of aspartate transaminase, alanine transaminase, lactate dehydrogenase, TNF-alpha, and IL-1beta were significantly lower in the FR-treated group than in the control group, and liver tissue blood flow was significantly higher in the FR-treated group than in the control group. Histopathologically, tissue damage was milder in the FR-treated group than in the control group. Both p38 MAPK and JNK were markedly phosphorylated after 30 minutes of reperfusion, and FR inhibited the phosphorylation of p38 MAPK without affecting the JNK. CONCLUSIONS: FR decreased serum TNF-alpha and IL-1beta levels and liver injury associated with the inhibition of p38 MAPK activation. These results suggest that inhibiting the activation of p38 MAPK may attenuate warm ischemia-reperfusion injury of the liver. PMID- 11894042 TI - Salvage of Swedish adjustable gastric band after explantation of an infected access port. PMID- 11894043 TI - Acalculous eosinophilic cholecystitis from herbal medicine: a review of adverse effects of herbal medicine in surgical patients. PMID- 11894044 TI - Who is Hawkeye? PMID- 11894045 TI - Supernumerary intravagal parathyroid hyperplasia. PMID- 11894046 TI - Can instrument contamination lead to tumor implantation? PMID- 11894047 TI - Imaging of endoleaks. AB - The endovascular repair of abdominal aortic aneurysms is gaining widespread acceptance worldwide. It relies on the exclusion of the aneurysm sac from arterial pressure/blood flow to reduce the pressure within it and therefore prevent the fatal complication of rupture. The presence of an endoleak is clear evidence that communication between the native circulation and the aneurysm sac persists. Unfortunately, direct measurement of the sac pressure is not a practical or safe method for routine detection or follow-up of endoleaks. Therefore, a fast, safe, sensitive, and reproducible method must be available. Although many imaging modalities have been and continue to be investigated, computed tomography angiography remains the gold standard. This article describes the various modalities used for the detection of endoleaks and discusses their imaging characteristics. PMID- 11894048 TI - Troubleshooting techniques for abdominal aortic aneurysm endograft placement: when things go wrong. AB - Since the Food and Drug Administration approved the Ancure (Guidant Corp, Menlo Park, CA) and AneuRx (Medtronic Corp, Minneapolis, MN) abdominal aortic aneurysm stent-grafts, there has been a tremendous increase in the number of stent-graft implantations, both in the United States and worldwide. Successful stent-graft deployment requires complex interventional skills and accurate preprocedure planning. Even the most skilled operator may experience intraoperative difficulties during graft deployment. There are intraprocedural complications that are common to all stent-grafts, as well as specific complications that are unique to the particular stent-graft being implanted. The first step to managing an intraprocedural complication is to be able to quickly recognize that a deployment difficulty has occurred. The interventionalist must then be ready to use common troubleshooting techniques to rectify or avoid complications of stent graft delivery. PMID- 11894049 TI - Diagnosis and management of type 2 endoleaks after endovascular aneurysm repair. AB - Endovascular repair is a major treatment advance in patients with large infrarenal abdominal aortic aneurysms. Since the FDA approved two commercial devices 2.5 years ago, over 40,000 patients have undergone this procedure in the United States. Although we have learned a great deal, more than a few mysteries relating to the long-term performance of these devices remain. This results in never-ending surveillance protocols searching for graft failure and aneurysm expansion. One of the especially contentious issues is the management of type 2 endoleaks. Unlike other endoleaks that are related to problems with the graft and/or fixation, this type of leak occurs in patients with properly functioning devices. This is why so much controversy exists about whether or not these patients must be treated. Some advocate "watchful-waiting" intervention only when there is aneurysm expansion. Others routinely treat patients with type 2 endoleaks in an attempt to prevent expansion. As with most controversial topics, if you look carefully, there is more agreement than disagreement between the two groups. In this review, we will first describe the methods used for endoleak diagnosis and treatment. We will then review our current endoleak treatment algorithm and explain its rationale for use. PMID- 11894050 TI - Postoperative management: type I and III endoleaks. AB - The purpose of this article is to help the reader understand the importance of imaging findings and treatment strategies for type I and III endoleaks. Although the appearance of these leaks on computed tomography can be somewhat unremarkable and similar in appearance to type II endoleaks, it is critically important for the treating physician to make the correct diagnosis, as these endoleak types signify an incompletely treated aneurysm. Once the diagnosis of a type I or III endoleak is made, the next step in treatment is to identify the cause of the endoleak. Incomplete initial graft expansion, further arterial dilation, endograft migration, component separation, and tears within the graft fabric are all possible causes of type I and III endoleaks. A combination of computed tomography, plain film radiography, and diagnostic angiography may be necessary to make the diagnosis and identify the underlying cause of the complication. Once all of these factors have been determined, a decision has to be made of whether the endoleak can be treated through additional endovascular means or if endovascular therapy has failed for the patient, making open surgical revision necessary to treat the aneurysm. Illustrative cases of all endoleak types and their treatments are the focus of this article. PMID- 11894051 TI - Postoperative management: buttock claudication and limb thrombosis. AB - As a result of endovascular repair of abdominal aortic aneurysms and the necessary associated adjunctive procedures, postoperative buttock claudication and limb thrombosis are complications that every physician who implants stent grafts should be able to recognize and treat. Whereas the presenting complaints of these complications can be quite obvious, the treatment of them may be not so simple. Studies have shown that 28% of patients who underwent embolization of one or both hypogastric arteries develop buttock claudication. Yet 78% of these affected patients spontaneously resolve their symptoms. Strategies to both minimize and successfully treat this complication are obviously of the utmost importance. Likewise, limb thrombosis can be easy to recognize, but treatment strategies and methods to limit this complication can be quite complex and remain somewhat controversial. One center was able to reduce their limb thrombosis rate from 17% to 0% through the use of intravascular ultrasound and aggressive adjunctive stenting. The purpose of this article is to first review the data concerning these complications and then to discuss treatment strategies that are designed to minimize and treat the actual complication. PMID- 11894052 TI - Techniques for hypogastric artery embolization. PMID- 11894055 TI - Slim pickings for silicon specialists. PMID- 11894054 TI - Applications matter. PMID- 11894056 TI - Helping hands for Arab science. PMID- 11894057 TI - Beyond the cloning debate. PMID- 11894059 TI - Geneticists get steamed up over public access to rice genome. PMID- 11894060 TI - NASA urged to play waiting game on Hubble's retirement. PMID- 11894062 TI - Radiologist in the picture for top job at NIH. PMID- 11894063 TI - Manchester merger to spawn research giant. PMID- 11894065 TI - Formidable catalogue puts army of ants online. PMID- 11894068 TI - Neutrino physics: picking up the pieces. PMID- 11894066 TI - Genomics firm aims to fill Asian gene gap. PMID- 11894069 TI - Arab science: blooms in the desert. PMID- 11894071 TI - Favouritism in physics? PMID- 11894070 TI - In risk assessment, one has to admit ignorance. PMID- 11894075 TI - Grammar: the barest essentials. PMID- 11894076 TI - Developmental biology: senseless motion. PMID- 11894077 TI - Cosmology: maintaining the standard. PMID- 11894079 TI - Signal transduction: molecular ticket to enter cells. PMID- 11894080 TI - Oceanography: an extra dimension to mixing. PMID- 11894081 TI - Neurobiology: the bitter-sweet taste of amino acids. PMID- 11894084 TI - Instant neural control of a movement signal. AB - The activity of motor cortex (MI) neurons conveys movement intent sufficiently well to be used as a control signal to operate artificial devices, but until now this has called for extensive training or has been confined to a limited movement repertoire. Here we show how activity from a few (7-30) MI neurons can be decoded into a signal that a monkey is able to use immediately to move a computer cursor to any new position in its workspace (14 degrees x 14 degrees visual angle). Our results, which are based on recordings made by an electrode array that is suitable for human use, indicate that neurally based control of movement may eventually be feasible in paralysed humans. PMID- 11894082 TI - Cell biology: a new view of photoreceptors. PMID- 11894085 TI - Sonography: dizygotic twin survival in early pregnancy. AB - It has been suggested that losses of twin conceptuses in very early pregnancy are high, and that for every liveborn twin pair there are a further 10-12 twin pregnancies that end up as a singleton birth. Here we show that in a group of women who had double-ovulated and conceived, the probability of the second egg also becoming fertilized and developing is 20-30% - which is comparable to the probability of conception and survival of a single conceptus. We conclude that the presence of one embryo does not affect the development of its twin. PMID- 11894086 TI - A velocity dipole in the distribution of radio galaxies. AB - The motion of our Galaxy through the Universe is reflected in a systematic shift in the temperature of the cosmic microwave background-because of the Doppler effect, the temperature of the background is about 0.1 per cent higher in the direction of motion, with a correspondingly lower temperature in the opposite direction. This effect is known as dipole anisotropy. If our standard cosmological model is correct, a related dipole effect should also be present as an enhancement in the surface density of distant galaxies in the direction of motion. The main obstacle to finding this signal is the uneven distribution of galaxies in the local supercluster, which drowns out the small cosmological signal. Here we report a detection of the expected cosmological dipole anisotropy in the distribution of galaxies. We use a survey of radio galaxies that are mainly located at cosmological distances, so the contamination from nearby clusters is small. When local radio galaxies are removed from the sample, the resulting dipole is in the same direction as the temperature anisotropy of the microwave background, and close to the expected amplitude. The result therefore confirms the standard cosmological interpretation of the microwave background. PMID- 11894087 TI - Electrical discharge from a thundercloud top to the lower ionosphere. AB - For over a century, numerous undocumented reports have appeared about unusual large-scale luminous phenomena above thunderclouds and, more than 80 years ago, it was suggested that an electrical discharge could bridge the gap between a thundercloud and the upper atmosphere. Since then, two classes of vertically extensive optical flashes above thunderclouds have been identified-sprites and blue jets. Sprites initiate near the base of the ionosphere, develop very rapidly downwards at speeds which can exceed 107 m s-1 (ref. 15), and assume many different geometrical forms. In contrast, blue jets develop upwards from cloud tops at speeds of the order of 105 m s-1 and are characterized by a blue conical shape. But no experimental data related to sprites or blue jets have been reported which conclusively indicate that they establish a direct path of electrical contact between a thundercloud and the lower ionosphere. Here we report a video recording of a blue jet propagating upwards from a thundercloud to an altitude of about 70 km, taken at the Arecibo Observatory, Puerto Rico. Above an altitude of 42 km-normally the upper limit for blue jets and the lower terminal altitude for sprites-the flash exhibited some features normally observed in sprites. As we observed this phenomenon above a relatively small thunderstorm cell, we speculate that it may be common and therefore represent an unaccounted for component of the global electric circuit. PMID- 11894088 TI - Formation of isomorphic Ir3+ and Ir4+ octamers and spin dimerization in the spinel CuIr2S4. AB - Inorganic compounds with the AB2X4 spinel structure have been studied for many years, because of their unusual physical properties. The spinel crystallographic structure, first solved by Bragg in 1915, has cations occupying both tetrahedral (A) and octahedral (B) sites. Interesting physics arises when the B-site cations become mixed in valence. Magnetite (Fe3O4) is a classic and still unresolved example, where the tendency to form ordered arrays of Fe2+ and Fe3+ ions competes with the topological frustration of the B-site network. The CuIr2S4 thiospinel is another example, well known for the presence of a metal-insulator transition at 230 K with an abrupt decrease of the electrical conductivity on cooling accompanied by the loss of localized magnetic moments. Here, we report the determination of the crystallographic structure of CuIr2S4 below the metal insulator transition. Our results indicate that CuIr2S4 undergoes a simultaneous charge-ordering and spin-dimerization transition-a rare phenomenon in three dimensional compounds. Remarkably, the charge-ordering pattern consists of isomorphic octamers of Ir83+S24 and Ir84+S24 (as isovalent bi-capped hexagonal rings). This extraordinary arrangement leads to an elegant description of the spinel structure, but represents an increase in complexity with respect to all the known charge-ordered structures, which are typically based on stripes, slabs or chequerboard patterns. PMID- 11894089 TI - Onset of Asian desertification by 22 Myr ago inferred from loess deposits in China. AB - The initial desertification in the Asian interior is thought to be one of the most prominent climate changes in the Northern Hemisphere during the Cenozoic era. But the dating of this transition is uncertain, partly because desert sediments are usually scattered, discontinuous and difficult to date. Here we report nearly continuous aeolian deposits covering the interval from 22 to 6.2 million years ago, on the basis of palaeomagnetic measurements and fossil evidence. A total of 231 visually definable aeolian layers occur as brownish loesses interbedded with reddish soils. This new evidence indicates that large source areas of aeolian dust and energetic winter monsoon winds to transport the material must have existed in the interior of Asia by the early Miocene epoch, at least 14 million years earlier than previously thought. Regional tectonic changes and ongoing global cooling are probable causes of these changes in aridity and circulation in Asia. PMID- 11894090 TI - Strong emission of methyl chloride from tropical plants. AB - Methyl chloride is the largest natural source of ozone-depleting chlorine compounds, and accounts for about 15 per cent of the present atmospheric chlorine content. This contribution was likely to have been relatively greater in pre industrial times, when additional anthropogenic sources-such as chlorofluorocarbons-were absent. Although it has been shown that there are large emissions of methyl chloride from coastal lands in the tropics, there remains a substantial shortfall in the overall methyl chloride budget. Here we present observations of large emissions of methyl chloride from some common tropical plants (certain types of ferns and Dipterocarpaceae), ranging from 0.1 to 3.7 microg per gram of dry leaf per hour. On the basis of these preliminary measurements, the methyl chloride flux from Dipterocarpaceae in southeast Asia alone is estimated at 0.91 Tg yr-1, which could explain a large portion of missing methyl chloride sources. With continuing tropical deforestation, natural sources of chlorine compounds may accordingly decrease in the future. Conversely, the abundance of massive ferns in the Carboniferous period may have created an atmosphere rich in methyl chloride. PMID- 11894091 TI - A Jurassic mammal from South America. AB - The Jurassic period is an important stage in early mammalian evolution, as it saw the first diversification of this group, leading to the stem lineages of monotremes and modern therian mammals. However, the fossil record of Jurassic mammals is extremely poor, particularly in the southern continents. Jurassic mammals from Gondwanaland are so far only known from Tanzania and Madagascar, and from trackway evidence from Argentina. Here we report a Jurassic mammal represented by a dentary, which is the first, to our knowledge, from South America. The tiny fossil from the Middle to Late Jurassic of Patagonia is a representative of the recently termed Australosphenida, a group of mammals from Gondwanaland that evolved tribosphenic molars convergently to the Northern Hemisphere Tribosphenida, and probably gave rise to the monotremes. Together with other mammalian evidence from the Southern Hemisphere, the discovery of this new mammal indicates that the Australosphenida had diversified and were widespread in Gondwanaland well before the end of the Jurassic, and that mammalian faunas from the Southern Hemisphere already showed a marked distinction from their northern counterparts by the Middle to Late Jurassic. PMID- 11894092 TI - Bacterial growth and primary production along a north-south transect of the Atlantic Ocean. AB - The oceanic carbon cycle is mainly determined by the combined activities of bacteria and phytoplankton, but the interdependence of climate, the carbon cycle and the microbes is not well understood. To elucidate this interdependence, we performed high-frequency sampling of sea water along a north-south transect of the Atlantic Ocean. Here we report that the interaction of bacteria and phytoplankton is closely related to the meridional profile of water temperature, a variable directly dependent on climate. Water temperature was positively correlated with the ratio of bacterial production to primary production, and, more strongly, with the ratio of bacterial carbon demand to primary production. In warm latitudes (25 degrees N to 30 degrees S), we observed alternating patches of predominantly heterotrophic and autotrophic community metabolism. The calculated regression lines (for data north and south of the Equator) between temperature and the ratio of bacterial production to primary production give a maximum value for this ratio of 40% in the oligotrophic equatorial regions. Taking into account a bacterial growth efficiency of 30%, the resulting area of net heterotrophy (where the bacterial carbon demand for growth plus respiration exceeds phytoplankton carbon fixation) expands from 8 degrees N (27 degrees C) to 20 degrees S (23 degrees C). This suggests an output of CO2 from parts of the ocean to the atmosphere. PMID- 11894093 TI - Hearing visual motion in depth. AB - Auditory spatial perception is strongly affected by visual cues. For example, if auditory and visual stimuli are presented synchronously but from different positions, the auditory event is mislocated towards the locus of the visual stimulus-the ventriloquism effect. This 'visual capture' also occurs in motion perception in which a static auditory stimulus appears to move with the visual moving object. We investigated how the human perceptual system coordinates complementary inputs from auditory and visual senses. Here we show that an auditory aftereffect occurs from adaptation to visual motion in depth. After a few minutes of viewing a square moving in depth, a steady sound was perceived as changing loudness in the opposite direction. Adaptation to a combination of auditory and visual stimuli changing in a compatible direction increased the aftereffect and the effect of visual adaptation almost disappeared when the directions were opposite. On the other hand, listening to a sound changing in intensity did not affect the visual changing-size aftereffect. The results provide psychophysical evidence that, for processing of motion in depth, the auditory system responds to both auditory changing intensity and visual motion in depth. PMID- 11894094 TI - Embryonic assembly of a central pattern generator without sensory input. AB - Locomotion depends on the integration of sensory information with the activity of central circuitry, which generates patterned discharges in motor nerves to appropriate muscles. Isolated central networks generate fictive locomotor rhythms (recorded in the absence of movement), indicating that the fundamental pattern of motor output depends on the intrinsic connectivity and electrical properties of these central circuits. Sensory inputs are required to modify the pattern of motor activity in response to the actual circumstances of real movement. A central issue for our understanding of how locomotor circuits are specified and assembled is the extent to which sensory inputs are required as such systems develop. Here we describe the effects of eliminating sensory function and structure on the development of the peristaltic motor pattern of Drosophila embryos and larvae. We infer that the circuitry for peristaltic crawling develops in the complete absence of sensory input; however, the integration of this circuitry into actual patterns of locomotion requires additional information from the sensory system. In the absence of sensory inputs, the polarity of movement is deranged, and backward peristaltic waves predominate at the expense of forward peristalsis. PMID- 11894095 TI - Cbl-CIN85-endophilin complex mediates ligand-induced downregulation of EGF receptors. AB - Cbl is a multi-adaptor protein involved in ligand-induced downregulation of receptor tyrosine kinases. It is thought that Cbl-mediated ubiquitination of active receptors is essential for receptor degradation and cessation of receptor induced signal transduction. Here we demonstrate that Cbl additionally regulates epidermal growth factor (EGF) receptor endocytosis. Cbl rapidly recruits CIN85 (Cbl-interacting protein of 85K; ref. 6) and endophilins (regulatory components of clathrin-coated vesicles) to form a complex with activated EGF receptors, thus controlling receptor internalization. CIN85 was constitutively associated with endophilins, whereas CIN85 binding to the distal carboxy terminus of Cbl was increased on EGF stimulation. Inhibition of these interactions was sufficient to block EGF receptor internalization, delay receptor degradation and enhance EGF induced gene transcription, without perturbing Cbl-directed receptor ubiquitination. Thus, the evolutionary divergent C terminus of Cbl uses a mechanism that is functionally separable from the ubiquitin ligase activity of Cbl to mediate ligand-dependent downregulation of receptor tyrosine kinases. PMID- 11894096 TI - The endophilin-CIN85-Cbl complex mediates ligand-dependent downregulation of c Met. AB - Ligand-dependent downregulation of tyrosine kinase receptors is a critical step for modulating their activity. Upon ligand binding, hepatocyte growth factor (HGF) receptor (Met) is polyubiquitinated and degraded; however, the mechanisms underlying HGF receptor endocytosis are not yet known. Here we demonstrate that a complex involving endophilins, CIN85 and Cbl controls this process. Endophilins are regulatory components of clathrin-coated vesicle formation. Through their acyl-transferase activity they are thought to modify the membrane phospholipids and induce negative curvature and invagination of the plasma membrane during the early steps of endocytosis. Furthermore, by means of their Src-homology 3 domains, endophilins are able to bind CIN85, a recently identified protein that interacts with the Cbl proto-oncogene. Cbl, in turn, binds and ubiquitinates activated HGF receptor, and by recruiting the endophilin-CIN85 complex, it regulates receptor internalization. Inhibition of complex formation is sufficient to block HGF receptor internalization and to enhance HGF-induced signal transduction and biological responses. These data provide further evidence of a relationship between receptor-mediated signalling and endocytosis, and disclose a novel functional role for Cbl in HGF receptor signalling. PMID- 11894097 TI - Involvement of receptor-interacting protein 2 in innate and adaptive immune responses. AB - Host defences to microorganisms rely on a coordinated interplay between the innate and adaptive responses of immunity. Infection with intracellular bacteria triggers an immediate innate response requiring macrophages, neutrophils and natural killer cells, whereas subsequent activation of an adaptive response through development of T-helper subtype 1 cells (TH1) proceeds during persistent infection. To understand the physiological role of receptor-interacting protein 2 (Rip2), also known as RICK and CARDIAK, we generated mice with a targeted disruption of the gene coding for Rip2. Here we show that Rip2-deficient mice exhibit a profoundly decreased ability to defend against infection by the intracellular pathogen Listeria monocytogenes. Rip2-deficient macrophages infected with L. monocytogenes or treated with lipopolysaccharide (LPS) have decreased activation of NF-kappaB, whereas dominant negative Rip2 inhibited NF kappaB activation mediated by Toll-like receptor 4 and Nod1. In vivo, Rip2 deficient mice were resistant to the lethal effects of LPS-induced endotoxic shock. Furthermore, Rip2 deficiency results in impaired interferon-gamma production in both TH1 and natural killer cells, attributed in part to defective interleukin-12-induced Stat4 activation. Our data reflect requirements for Rip2 in multiple pathways regulating immune and inflammatory responses. PMID- 11894103 TI - Cold atoms and quantum control. AB - This overview prefaces a collection of Insight review articles on the physics and applications of laser-cooled atoms. I will cast this work into a historical perspective in which laser cooling and trapping is seen as one of several research directions aimed at controlling the internal and external degrees of freedom of atoms and molecules. PMID- 11894104 TI - Bose-Einstein condensation of atomic gases. AB - The early experiments on Bose-Einstein condensation in dilute atomic gases accomplished three long-standing goals. First, cooling of neutral atoms into their motional ground state, thus subjecting them to ultimate control, limited only by Heisenberg's uncertainty relation. Second, creation of a coherent sample of atoms, in which all occupy the same quantum state, and the realization of atom lasers - devices that output coherent matter waves. And third, creation of a gaseous quantum fluid, with properties that are different from the quantum liquids helium-3 and helium-4. The field of Bose-Einstein condensation of atomic gases has continued to progress rapidly, driven by the combination of new experimental techniques and theoretical advances. The family of quantum degenerate gases has grown, and now includes metastable and fermionic atoms. Condensates have become an ultralow-temperature laboratory for atom optics, collisional physics and many-body physics, encompassing phonons, superfluidity, quantized vortices, Josephson junctions and quantum phase transitions. PMID- 11894106 TI - Quantum encounters of the cold kind. AB - Since the introduction of laser-cooling techniques for neutral atoms in the early 1980s, the study of collisional interactions between atoms and molecules has been extended to the regime of ultracold temperatures. With nanokelvin temperatures now attainable, our ability to probe the interactions, both experimentally and theoretically, has also progressed. Understanding of the subtle and often highly quantum-mechanical effects that are manifest at such low energies has advanced to the point where new precision measurements are matched by highly accurate theoretical calculations. Low-energy phenomena such as Bose-Einstein condensation and the photoassociation of atoms into bound molecules are now accurately described with no free parameters. PMID- 11894105 TI - Nonlinear and quantum atom optics. AB - Coherent matter waves in the form of Bose-Einstein condensates have led to the development of nonlinear and quantum atom optics - the de Broglie wave analogues of nonlinear and quantum optics with light. In nonlinear atom optics, four-wave mixing of matter waves and mixing of combinations of light and matter waves have been observed; such progress culminated in the demonstration of phase-coherent matter-wave amplification. Solitons represent another active area in nonlinear atom optics: these non-dispersing propagating modes of the equation that governs Bose-Einstein condensates have been created experimentally, and observed subsequently to break up into vortices. Quantum atom optics is concerned with the statistical properties and correlations of matter-wave fields. A first step in this area is the measurement of reduced number fluctuations in a Bose-Einstein condensate partitioned into a series of optical potential wells. PMID- 11894098 TI - RICK/Rip2/CARDIAK mediates signalling for receptors of the innate and adaptive immune systems. AB - The immune system consists of two evolutionarily different but closely related responses, innate immunity and adaptive immunity. Each of these responses has characteristic receptors-Toll-like receptors (TLRs) for innate immunity and antigen-specific receptors for adaptive immunity. Here we show that the caspase recruitment domain (CARD)-containing serine/threonine kinase Rip2 (also known as RICK, CARDIAK, CCK and Ripk2) transduces signals from receptors of both immune responses. Rip2 was recruited to TLR2 signalling complexes after ligand stimulation. Moreover, cytokine production in Rip2-deficient cells was reduced on stimulation of TLRs with lipopolysaccharide, peptidoglycan and double-stranded RNA, but not with bacterial DNA, indicating that Rip2 is downstream of TLR2/3/4 but not TLR9. Rip2-deficient cells were also hyporesponsive to signalling through interleukin (IL)-1 and IL-18 receptors, and deficient for signalling through Nod proteins-molecules also implicated in the innate immune response. Furthermore, Rip2-deficient T cells showed severely reduced NF-kappaB activation, IL-2 production and proliferation on T-cell-receptor (TCR) engagement, and impaired differentiation to T-helper subtype 1 (TH1) cells, indicating that Rip2 is required for optimal TCR signalling and T-cell differentiation. Rip2 is therefore a signal transducer and integrator of signals for both the innate and adaptive immune systems. PMID- 11894107 TI - Optical frequency metrology. AB - Extremely narrow optical resonances in cold atoms or single trapped ions can be measured with high resolution. A laser locked to such a narrow optical resonance could serve as a highly stable oscillator for an all-optical atomic clock. However, until recently there was no reliable clockwork mechanism that could count optical frequencies of hundreds of terahertz. Techniques using femtosecond laser frequency combs, developed within the past few years, have solved this problem. The ability to count optical oscillations of more than 1015 cycles per second facilitates high-precision optical spectroscopy, and has led to the construction of an all-optical atomic clock that is expected eventually to outperform today's state-of-the-art caesium clocks. PMID- 11894108 TI - Quantum information processing with atoms and photons. AB - Quantum information processors exploit the quantum features of superposition and entanglement for applications not possible in classical devices, offering the potential for significant improvements in the communication and processing of information. Experimental realization of large-scale quantum information processors remains a long-term vision, as the required nearly pure quantum behaviour is observed only in exotic hardware such as individual laser-cooled atoms and isolated photons. But recent theoretical and experimental advances suggest that cold atoms and individual photons may lead the way towards bigger and better quantum information processors, effectively building mesoscopic versions of 'Schrodinger's cat' from the bottom up. PMID- 11894099 TI - An amino-acid taste receptor. AB - The sense of taste provides animals with valuable information about the nature and quality of food. Mammals can recognize and respond to a diverse repertoire of chemical entities, including sugars, salts, acids and a wide range of toxic substances. Several amino acids taste sweet or delicious (umami) to humans, and are attractive to rodents and other animals. This is noteworthy because L-amino acids function as the building blocks of proteins, as biosynthetic precursors of many biologically relevant small molecules, and as metabolic fuel. Thus, having a taste pathway dedicated to their detection probably had significant evolutionary implications. Here we identify and characterize a mammalian amino-acid taste receptor. This receptor, T1R1+3, is a heteromer of the taste-specific T1R1 and T1R3 G-protein-coupled receptors. We demonstrate that T1R1 and T1R3 combine to function as a broadly tuned L-amino-acid sensor responding to most of the 20 standard amino acids, but not to their D-enantiomers or other compounds. We also show that sequence differences in T1R receptors within and between species (human and mouse) can significantly influence the selectivity and specificity of taste responses. PMID- 11894109 TI - p53 Gene mutation and genetic instability in superficial multifocal esophageal squamous cell carcinoma. AB - Tumor multicentricity is occasionally observed in esophageal squamous cell carcinoma (SCC). We studied five surgically resected superficial multifocal esophageal SCCs for p53 gene mutation and genetic instability, using DNA extracted from microdissected areas. A total of 38 target areas (TAs) were analyzed in SCC, dysplasia, basal cell hyperplasia (BCH) and normal squamous epithelium. Analysis of the replication error (RER) at 10 microsatellite loci showed microsatellite instability in all TAs, as well as in normal squamous epithelium. p53 gene mutation was identified in 28.9% (11/38 TAs). All cases showed a common missense mutation in exon 8 at codon 273 (CGT-->CAT, Arg-->His), which was DNA contact mutation in the S10 beta strand. In association with microsatellite alterations, 7 of 9 TAs with p53 mutation in exon 8 at codon 273 also showed loss of heterozygosity (LOH) of p53 gene. LOH of p53 gene was detected in 83.8% (31/37 TAs). LOH at D2S123 on 2p16 near MSH2 gene and at D3S1611 on 3p22 near MLH1 gene was detected in 65.4% (17/26) and 71.4% (10/14) TAs, respectively. Frequencies of LOH at p53 and D2S123 were similar in non cancerous areas and SCCs. LOH of p53 and D2S123 were found in 50% (5/10 TAs) of non-cancerous areas and 60% (9/15 TAs) of SCCs. Our results suggest that genetic instability induces esophageal tumor multicentricity, and that p53 gene contact mutation together with LOH are early events of the multistage carcinogenesis of multifocal primary esophageal SCC. PMID- 11894110 TI - Effects of herbal preparation Equiguard on hormone-responsive and hormone refractory prostate carcinoma cells: mechanistic studies. AB - The Equiguard is a dietary supplement comprised of standardized extracts from nine herbs, respectively, Herba epimedium brevicornum Maxim (stem and leaves), Radix morindae officinalis (root), Fructus rosa laevigatae michx (fruit), Rubus chingii Hu (fruit), Schisandra chinensis (Turz.) Baill (fruit), Ligustrum lucidum Ait (fruit), Cuscuta chinensis Lam (seed), Psoralea corylifolia L. (fruit), and Astragalus membranaceus (Fisch.) Bge (root). This proprietary product, formulated according to Chinese traditional medicinal concepts, is aimed at restoring harmony in the of the kidney, an organ which Chinese medicinal principles consider to be vital for invigorating as well as maintaining balance of the entire urological system. As the prostate is an integral component of the urological system, we performed in vitro studies to test the effects of ethanol extracts of Equiguard to modulate prostate growth and gene expression. These studies used prostate cancer cells mimicking the androgen dependent (AD) and androgen-independent (AI) states of prostate carcinogenesis. Results show that Equiguard significantly reduced cancer cell growth, induced apoptosis, suppressed expression of the androgen receptor (AR) and lowered intracellular and secreted prostate specific antigen (PSA), and almost completely abolished colony forming abilities of prostate cancer cells. These data support the interpretation that this herbal formulation contains ingredients that collectively may be efficacious in preventing or treating AD and AI prostate carcinoma. The anti-prostatic activities of Equiguard may stem from its complex composition capable of targeting multiple signal transduction/metabolic pathways, to effectively correct, counteract or circumvent the impaired or dysfunctional mechanisms accompanying different stages of prostate carcinogenesis. PMID- 11894112 TI - A comparative guide to gene prediction tools for the bioinformatics amateur. AB - Several hundred programs using different algorithms have been designed to predict individual coding features within any genomic sequence, but none of these tools covers all aspects of a gene or is 100% accurate in its prediction. Automated simultaneous processing of the results from a number of these programs minimizes the chance of a false positive prediction and quickly generates integrated data. We report here on the analysis of two known genes in 5 and 25 kb segments of genomic sequence using four genome annotation packages, NIX, RUMMAGE, Genotator and EMBOSS. Gene predictions were confirmed using cDNA sequences and a comparison was made between the packages. This study showed a similarity in the ability of NIX, RUMMAGE and Genotator to predict well-characterised genes and basic structures, but poor exon prediction for a small, 3 exon gene. However, the BLAST subprograms of all three packages correctly identified the 3 exons. In addition, EST BLAST subprograms identified a previously undescribed, possible 5' untranslated exon for the smaller gene and a number of putative alternatively spliced exons in the larger gene. Overall, NIX was found to be the most user friendly package, in terms of easy access to databases and the interactive graphical display of results. PMID- 11894111 TI - Clinical relevance of fusion images using (18)F-2-fluoro-2-deoxy-D-glucose positron emission tomography in local recurrence of rectal cancer. AB - Local recurrence of rectal cancer is a critical issue. Anatomical images, such as computed tomography (CT) or magnetic resonance imaging (MRI), are sometimes insufficient for preoperative evaluation. A useful modality for diagnosis of local recurrence of rectal cancer is (18)F-2-fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET), but it does not give adequate information about anatomy. To utilize the advantages of these techniques, this study was done to validate the accuracy of using fused images and their usefulness in the decision making for surgical intervention of local recurrence of rectal cancer by directly comparing the fused images with resected specimens. PET and CT/MRI were performed for patients suspected of local recurrence of rectal cancer (n=4). PET image data were re-calculated to fit CT/MRI images and manually superimposed on the anatomical images. Fusion images were compared with resected specimens. Radical operation was carried out for three patients. Fusion images provided information on precise tumor location, and extent of tumor invasion as well as the diagnosis of tumor recurrence. All patients underwent curative operation with negative surgical margins, and the information provided by the fusion images was confirmed by comparison with resected specimens. In all cases, preoperative evaluation of tumor recurrence with fusion images provided more useful clinical information for the management of patients than the anatomical images alone. PET images, when combined with MRI or CT, may prove to be a useful adjunct in the management of patients being evaluated for resection of local recurrence of rectal cancer. PMID- 11894113 TI - Fetal ontogeny and tumor expression of the early thymic antigen UN1. AB - UN1 antigen (Ag), a 100-120 kDa sialoglycoprotein, was initially identified on immature thymocytes (CD3(dim)), a small subpopulation of CD4(+) peripheral blood T-lymphocytes, on leukemic T-cell lines and in fetal thymus. Biochemical analysis of the Ag has identified molecular features that are characteristic of cell membrane-associated mucin-like glycoproteins. To investigate the biological role and the potential usefulness of the Ag, we have more extensively studied the pattern of UN1 Ag expression in a panel of fetal tissues, at different gestational ages, and on adult normal and tumor specimens. In the fetal samples examined by immunohistochemistry, including intestine, liver, lung and adrenal gland, we found that UN1 Ag is widely expressed during early stages of fetal development and down-regulated during ontogenesis. Very poor or not detectable expression of UN1 Ag was found at late gestational age. Immunohistochemical, Western blot and flow cytometric analysis of a panel of normal adult tissues and benign lesions failed to find Ag expression, whereas UN1 Ag was highly detectable in a variety of cancer specimens from breast, lung, gastrointestinal, gynaecological malignancies and melanomas. Based on these data UN1 Ag, for the wide expression on fetal tissues, the down-regulation during ontogeny and the re expression in cancer cells, may be considered a novel oncofetal Ag of interest for biological investigation and clinical applications. PMID- 11894115 TI - An evaluation of the prognostic significance of vascular endothelial growth factor in node positive primary breast carcinoma. AB - Angiogenesis is intimately related to the growth and progression of tumours and must be induced to facilitate growth beyond a minimum size. It has been implicated in the development of metastases and survival in breast carcinoma. VEGF is a cytokine that plays an important role in angiogenesis. Its expression is increased in solid tumours during induction of angiogenesis and it has been implicated as a prognostic marker in patients with node negative breast carcinoma. We studied VEGF expression, in a series of patients with node positive breast carcinoma and examined histopathological parameters of the tumour and the prognostic value of VEGF expression. Specimens from 108 cases of node positive breast cancer were stained for VEGF using an antibody suitable for use on formalin fixed tissue. VEGF staining was cytoplasmic and was scored by intensity and the percent positive cells. Patients with positive VEGF staining (n=48) were compared with patients with negative VEGF staining (n=60). Demographic criteria were similar in both groups. Only one (12%) patient with lobular carcinoma and one (14%) patient with medullary carcinoma expressed VEGF compared with 46 (49%) patients with ductal carcinoma (NOS). DCIS was present in 60 tumours. There was a strong correlation between staining in DCIS and the adjacent invasive tumours. There was no significant association between VEGF staining and T stage, tumour size or the number of positive lymph nodes. VEGF expression had no prognostic significance either for disease-free or overall survival in patients with node positive disease. This study failed to support a role for VEGF as a prognostic marker in patients with node positive breast carcinoma. PMID- 11894114 TI - Expression of PLAG1 and HMGIC proteins and fusion transcripts in radiation associated pleomorphic adenomas. AB - Extensive cytogenetic investigations of pleomorphic adenomas of the salivary glands have unequivocally demonstrated that they are cytogenetically monoclonal and are characterized by a high frequency of tumor specific chromosome abnormalities involving in particular chromosome bands 3p21, 8q12 and 12q14-15. Here we show that two radiation-associated and cytogenetically polyclonal adenomas without gross rearrangements of these breakpoints show simultaneous overexpression of the PLAG1 and HMGIC genes, i.e. the target genes of the 8q12 and 12q14-15 rearrangements in sporadic adenomas. In addition, one of the tumors expressed a cryptic CTNNB1-PLAG1 fusion transcript. Our findings strongly suggest that identical or very similar molecular mechanisms are operating during adenoma tumorigenesis irrespective of whether the tumors are cytogenetically polyclonal or whether they have non-random, tumor specific abnormalities. Cytogenetically polyclonal adenomas are thus most likely also of monoclonal origin. PMID- 11894116 TI - Bax-induction alone is sufficient to activate apoptosis cascade in wild-type Bax bearing K562 cells, and the initiation of apoptosis requires simultaneous caspase activation. AB - It has been reported that expression of Bax by Tet-On system induces apoptosis in Jurkat cells. The parental Jurkat cells have mutation of Bax gene and do not express Bax protein. Wild-type Bax-bearing cells express endogenous Bax protein and it is still unclear whether overexpression of Bax alone can sufficiently induce apoptosis in these cells in the absence of any cytotoxic stimulus. To investigate this, wild-type Bax-bearing K562 cells were transfected with Tet-On Bax-inducible system (pTet-On and pTRE-Bax plasmids), and Bax-inducible stable cell lines were established. Overexpression of Bax in wild-type Bax-bearing K562 cells without any cyctotoxic signal resulted in increase of apoptotic cells, caspase-3 activation, mitochondrial release of cytochrome c, and mitochondrial membrane potential change. Western blotting and confocal microscopy revealed that overexpression of Bax was detected in mitochondria. A pancaspase inhibitor, zVAD fmk, which has no effect on mitochondrial cytochrome c release and mitochondrial membrane potential change inhibited the apoptotic events in the presence of overexpressed Bax in mitochondria. These findings suggest that Bax protein, when present above a threshold level, is sufficient to trigger an apoptosis cascade, and its initiation requires simultaneous caspase activation probably not mediated by mitochondrial cytochrome c release and mitochondrial membrane potential change in K562 cells. PMID- 11894117 TI - Expression of interleukin-10 is inversely correlated with distant metastasis of renal cell carcinoma. AB - Interleukin-10 (IL-10) is an immunomodulative cytokine produced by T cells, B cells, and monocytic cells. The significance of IL-10 and IL-10R expression in renal cell carcinoma (RCC) has not been well characterized. In this study, we assessed the correlation between IL-10 gene expression and its pathological significance in 44 RCC specimens. The patients with IL-10 gene expression showed a decreased incidence of metastasis compared to those without IL-10 gene expression. The expression of IL-10 was correlated with vascular endothelial growth factor (VEGF)-A gene expression. These findings suggest that IL-10 gene expression plays some roles in the metastasis of RCC. PMID- 11894118 TI - Alternately spliced luteinizing hormone/human chorionic gonadotropin receptor mRNA in human breast epithelial cells. AB - Luteinizing hormone (LH)/human chorionic gonadotropin (hCG) bind to a common transmembrane glycoprotein receptor, which is a member of the G protein-coupled receptor family. In human, the LH/hCG receptor gene is composed of 11 exons and 10 introns and its coding region is over 60 kb long. Human chorionic gonadotropin is a glycoprotein hormone containing a cystine-knot folding motif that is found in peptide growth factors known to activate the expression of homeogenes. In the present work we present evidence that hCG down-regulates all the three transcripts of HOXA1 at early stages of hCG treatment in the immortalized human breast epithelial cells (MCF-10F), whereas HOXA1-S1, the largest transcript, as well as HOXA1-S3, the smallest transcript, were up-regulated in the cancer cell lines MDA-MB-231 and MCF-7, respectively. This divergent reaction of hCG was associated with the pattern of LH/hCG receptors and splicing forms expression in human breast epithelial cells. MCF-10F cells expressed the full-length (1191 bp) compared with the cancer-derived cells MCF-7 and MDA-MB-231 that was weakly or not expressed. Isoform 1 (1117 bp) was silent in MCF-10F and expressed weakly in the cancer cells. The isoforms 2 (1006 bp) and 3 (932 bp) of LH/hCG gene receptor were silent in all the cell lines, whereas isoforms 4 (892 bp), 6 (626 bp) and 7 (441 bp) were silent in MCF-10F cells and expressed in the cancer cell lines. Instead isoform 5 (707 bp) showed in the three cell lines the strongest expression in MCF-10F cells. This difference in the expression of alternate splicing of LH/hCG receptor mRNA among the MCF-10F, MCF-7 and MDA-MB-231 cells, may explain the divergent response of these cells to HOXA1 activation by hCG. PMID- 11894119 TI - Molecular cloning and characterization of bovine bgl-1, a novel family member of WD-40 repeat-containing lethal giant larvae tumor suppressor genes. AB - In this study, we have isolated a bovine homologue bgl-1 of lethal giant larvae (lgl) tumor suppressor oncogene from bovine brain by RT-PCR using primers designed based on the conserved sequences for lgl family members. The sequence analysis showed that the bgl-1 encodes a 1,036 amino acid polypeptide with the molecular weight of approximately 112 kDa containing a domain characteristic of WD-40 proteins. The amino acid sequence of bgl-1 showed a homology of 98.3 and 87.3% identity to that of mouse and human, respectively. Northern blot analysis showed that bgl-1 was highly expressed in brain, ovary and testis, with moderate expression in liver, uterus, lung and kidney. This suggests that the bgl-1 may play essential roles in each of these organs. The complementation analysis revealed that the bovine bgl-1 partially restored the Na+ tolerance in the absence of yeast lgl homologue, suggesting that bgl-1 is a bovine homologue of the lgl family. PMID- 11894120 TI - Antisense anti-MDM2 mixed-backbone oligonucleotides enhance therapeutic efficacy of topoisomerase I inhibitor irinotecan in nude mice bearing human cancer xenografts: In vivo activity and mechanisms. AB - Antisense oligonucleotides have been investigated as anticancer agents administered alone or in combination with conventional chemotherapeutics. In the present study, we demonstrated synergistic effects between anti-MDM2 antisense oligonucleotides and the clinically used anticancer agent irinotecan, using nude mouse models of human colon cancers (LS174T and DLD-1). Surprisingly, a 5-base mismatch oligonucleotide also showed similar effects. To elucidate the underlying mechanisms, in vitro and in vivo pharmacokinetic and pharmacodynamic studies were performed. In LS174T cells, the antisense oligonucleotide, but not the mismatch oligonucleotide, specifically inhibited MDM2 expression, resulting in a significant increase in irinotecan-associated p53 activation and p21 induction. In DLD-1 cells, the antisense oligonucleotide specifically inhibited MDM2 expression, resulting in a significant increase in irinotecan-associated p21 induction although mutant p53 levels remained unchanged. Both oligonucleotides increased tissue uptake of irinotecan and the conversion of irinotecan to its active metabolite SN-38. These results suggest that oligonucleotides have a role in irinotecan metabolism and action, providing a basis for future development of antisense oligonucleotides as a sensitizer for irinotecan-based therapy. PMID- 11894121 TI - Lamin B, caspase-3 activity, and apoptosis induction by a combination of HMG-CoA reductase inhibitor and COX-2 inhibitors: a novel approach in developing effective chemopreventive regimens. AB - Apoptosis plays a central role in tumor development and it has been hypothesized that lack/failure of apoptosis leads to the development of tumors, including colon tumors. Thus, induction of apoptosis in tumor cells is an effective approach to the regulation of tumor growth. It has been shown by us and other investigators that various chemopreventive agents induce apoptosis and inhibit tumor growth. Identification of agents or combinations of agents that induce tumor cell apoptosis guides the development of novel agents for colon cancer treatment. Experiments were designed to assess the effectiveness of lovastatin, a 3-hydroxy-3-methyl glutaryl-CoA reductase inhibitor, and celecoxib a cyclooxygenase-2 inhibitor, individually or in combination on the induction of apoptosis in human HT-29 colon cancer cells. In addition, we studied the modulatory effect of lovastatin and celecoxib on lamin B levels, caspase-3 activity and expression in relationship to apoptosis in colon cancer cell lines. HT-29 cells exposed to various subtoxic levels of lovastatin or celecoxib or a combination of both were analyzed for apoptosis (by DAPI method), caspase-3 expression (immunoblot analysis) and caspase-3 activity (fluorimetric method). We found that: i) pretreatment with lovastatin (5-30 microM) induces apoptosis in HT 29 cells significantly only at high concentrations (> or = 20 microM) but not at low dose levels; ii) similarly, pretreatment with celecoxib produced apoptosis in colon cancer cells at high concentrations only (> or = 75 microM); iii) caspase-3 protein expression was moderately altered by the treatment with lovastatin or celecoxib at lower concentrations; however, a significant increase (1.6 to 4 fold) in caspase-3 expression and activity was found in HT-29 cells exposed with 20-25 microM lovastatin and/or 5-125 microM celecoxib and iv) importantly, in tumor cells exposed to low doses of (5 or 10 microM) lovastatin, combined with 25 75 microM of celecoxib, apoptosis induction rose 2.5 to 10-fold, caspase-3 expression was 2.3 to 8-fold higher, and enzyme activities were 1.5 to 5.5-fold elevated. This effect was highly synergistic and dose-dependent. Lamin B levels were significantly increased in a dose-dependent manner in cells treated with lovastatin but no such effect was observed with celecoxib. These results indicate that agents with different modes of action when applied in combinations will induce apoptosis synergistically by enhancing caspase-3 activities. These findings further support the hypothesis that HMGCo-R and COX-2 activities play important roles in apoptosis and regulation of apoptosis by selective agents such as lovastatin and celecoxib would provide effective strategies for the prevention of colon cancer. PMID- 11894122 TI - Fibroblasts capture cathepsin D secreted by breast cancer cells: possible role in the regulation of the invasive process. AB - Breast cancer cells oversecrete the lysosomal peptidase cathepsin D as a pro enzyme. In this study, we assessed the effect of media conditioned by MRC-5 fibroblasts or MCF-7/6 breast cancer cells on cathepsin D (CD) production and secretion by these two cell types. We also considered the influence of estrogens and matrix components (type I or type IV collagen, or Matrigel) on the expression and activity of CD produced by breast cancer cells of different invasive potentials (MCF-7/AZ, MCF-7/6, MDA-MB-231). In our system, fibroblasts conditioned medium does not significantly affect CD levels produced and secreted by the MCF-7/6 cells. However, we found that fibroblasts are able to capture the pro-CD secreted by these tumor cells by a mannose 6-phosphate-dependent process. We also found a positive correlation between the proportion of extracellular CD levels and the invasive potentials of the tumor cell types considered. If estrogens are able to upregulate CD production and secretion by receptor-positive cells, it is not the case of extracellular matrix components. On the other hand, our results indicate that matrix components are able to influence the distribution of the different CD forms in and out of the cells. Our data suggest that tumor fibroblasts, by enhancing their intracellular CD levels, could assist cancer cells in the digestion of extracellular matrix during the invasion of tissues. PMID- 11894123 TI - Expression of p57(KIP2) in hepatocellular carcinoma: relationship between tumor differentiation and patient survival. AB - Cyclins, cyclin-dependent kinases (CDK), and CDK inhibitors (CKI) are frequently altered in neoplasm. p57(KIP2) is a member of the KIP (kinase inhibitory protein) family of CKI and is a potential tumor suppressor gene. p27(KIP1) is the most extensively studied KIP family member with respect to the clinical significance in human neoplasms. However, the clinical significance of p57(KIP2) expression in patients with human cancers, including hepatocellular carcinoma (HCC), remains unknown. This study examined whether p57(KIP2) expression has any impact on clinical behavior of HCC including prognosis. We examined an expression of p57(KIP2) by immunohistochemistry in 101 cases of various liver diseases, including 59 HCC. The p57(KIP2) expression in HCC was analyzed in association with the pathohistologic stage, differentiation, proliferating cell nuclear antigen (PCNA) expression status and several histopathologic factors of possible prognostic value, and patient survival. Immunohistochemical analysis revealed the frequent loss of p57(KIP2) in HCC, especially in moderately and poorly differentiated HCC. By Kaplan-Meier analysis, overall survival was significantly correlated with p57(KIP2) expression and PCNA, and multivariate analysis showed p57(KIP2) was an independent prognostic factor. When the status of p57(KIP2) and PCNA were combined, cases positive for p57(KIP2) and with a low expression of PCNA had a significantly better prognosis than those negative for p57KIP2 and/or with a high expression of PCNA. These data indicate that loss of p57(KIP2) is a frequent event in HCC, especially in poorly differentiated HCC, suggesting that p57(KIP2) might play a role in the differentiation of HCC. In addition, p57(KIP2) expression status is an independent prognostic factor for patients with HCC, and the loss of p57(KIP2) is correlated with poor prognosis. New therapeutic options might be provided by the combination of the loss of p57(KIP2) and expression of PCNA. PMID- 11894124 TI - Expression of WRCH1 in human cancer and down-regulation of WRCH1 by beta estradiol in MCF-7 cells. AB - Secreted-type glycoprotein WNTs bind to seven-transmembrane-type WNT receptors encoded by Frizzled genes (FZD1-FZD10) to transduce signals to the beta-catenin- TCF pathway, the JNK pathway, or the Ca(2+)-releasing pathway. Wrch1 gene is a down-stream target gene of Wnt1 in C57MG cells, and encodes a Cdc42-related GTPase with the potential to activate the JNK pathway. Here, we isolated human WRCH1 cDNAs (accession no. AB074878) from gastric cancer cell lines OKAJIMA, MKN7, MKN28, MKN45, MKN74, and KATO-III, all of which showed a nucleotide substitution (343 C-->T) without amino-acid substitution compared with WRCH1 cDNA isolated by another group. WRCH1 gene, consisting of at least 3 exons, was mapped to human chromosome 1q42.11-q42.3 by using bioinformatics. WRCH1 mRNA was more highly expressed in corpus callosum, hippocampus, cerebral cortex, and also in several parts within adult brain than in other normal tissues including stomach, pancreas, and placenta. Amounts of WRCH1 mRNA in 40 human cancer cell lines were lower than that in normal stomach, pancreas, or placenta. WRCH1 mRNA was significantly up-regulated in 4 cases of primary kidney tumors, 1 case each of primary colon, gastric, breast, ovarian, and uterus cancer. On the other hand, WRCH1 mRNA was significantly down-regulated in 6 cases of colon tumors, 2 cases of primary kidney cancer and breast cancer. Expression of WRCH1 mRNA was down regulated by beta-estradiol in MCF-7 cells. This is the first report on comprehensive expression analyses on WRCH1 mRNA. PMID- 11894125 TI - Molecular cloning and expression of proto-oncogene FRAT1 in human cancer. AB - FRAT1 and FRAT2 genes, clustered in human chromosome 10q24, are human homologues to mouse proto-oncogene Frat1, which promotes carcinogenesis through activation of the WNT - beta-catenin - TCF signaling pathway. FRAT1 and FRAT2 mRNAs are up regulated together in a gastric cancer cell line TMK1, and also in 2 out of 10 cases of primary gastric cancer. Here, we isolated FRAT1 cDNA (AB074890), which showed two amino-acid substitutions (Gln57X and His58Asp) compared with human FRAT1 cDNA previously reported by another group (U58975). The Gln57-His58 FRAT1 allele isolated in this study was also identified in human genome draft sequences. FRAT1 mRNA was almost ubiquitously expressed in human pancreatic cancer cell lines. Expression level of FRAT1 mRNA was relatively higher in esophageal cancer cell lines TE2, TE3, TE4, a cervical cancer cell line SKG-IIIa, and breast cancer cell lines MCF-7 and T-47D. Expression level of FRAT1 mRNA was not significantly changed after all-trans retinoic-acid treatment in NT2 cells with the potential of neuronal differentiation. Expression of FRAT1 mRNA in MCF-7 cells derived from breast cancer was down-regulated by beta-estradiol. This is the first report on isolation of FRAT1 cDNA derived from the more common FRAT1 allele, and also on regulation of FRAT1 mRNA in human cancer cells. PMID- 11894126 TI - Scarff-Bloom-Richardson (SBR) grading: a pleiotropic marker of chemosensitivity in invasive ductal breast carcinomas treated by neoadjuvant chemotherapy. AB - The Scarff-Bloom-Richardson (SBR) grade, an important prognostic factor in breast cancer, was also associated with cell proliferation, a consistent indicator of response to chemotherapy. The determination of an association between SBR grade and responsiveness would be clinically useful. We explored the influence of SBR grade on response to neoadjuvant chemotherapy in patients with invasive ductal breast carcinoma. The present study centered on 431 patients registered onto one of four prospective phase II trials. SBR grading was performed according to the Elston method on needle core biopsies prospectively collected prior to treatment from 290 patients and on residual tumour at surgery from 171 patients. The post operative grades were then compared with those obtained at diagnosis. Univariate and multivariate analysis were used to evaluate the significance of SBR grade on response to neoadjuvant chemotherapy. Both statistical analysis revealed that SBR grade III tumours responded better to neoadjuvant treatment than SBR grade I (p<10(-6)). None of the other patient and tumour characteristics tested correlated with response. Moreover, tumour responsiveness was significantly related to changes of the SBR grade (p=7 x 10(-3)). As a conclusion, we showed that SBR grade is a strong predictive factor of response to induction chemotherapy in breast cancer, independently of the type of regimen used. The association between evolution of the histological grade following chemotherapy and response to treatment may prove valuable for clinicians as they make their decision regarding patient therapy. PMID- 11894127 TI - Decrease in growth factor receptors after treatment with serine protease inhibitor ONO-3403. AB - FOY-305 is a synthetic serine protease inhibitor and ONO-3403 is a derivative with a higher protease-inhibitory activity. The growth-suppressive effects of ONO 3403 were more prominent in Ha-ras-transformed NIH3T3 (ras-NIH) cells than in non transformed NIH3T3 cells. After treatment of ras-NIH cells with ONO-3403 at 100 200 microg/ml, the percentage of cells found in G(1) phase decreased and, concomitantly, that in S phase increased. Molecular events caused by ONO-3403 were investigated by Western blot analysis using anti-phosphotyrosine antibody. The results showed a marked decrease in the tyrosine phosphorylation level of a 180-kDa protein after treatment with ONO-3403. This 180-kDa phosphotyrosine containing molecule which was tentatively designated pY-p180 might be platelet derived growth factor (PDGF) receptor since addition of PDGF to serum-starved NIH3T3 cells induced a marked tyrosine phosphorylation of the same size within 5 min. This was further confirmed by immunoprecipitation of cell extract with anti PDGF-receptor antibody followed by Western blot analysis using anti phosphotyrosine antibody. Treatment of T.Tn human esophageal carcinoma cells with ONO-3403 caused also decrease in pY-p180, which appeared to be epidermal growth factor receptor. Thus, ONO-3403 may induce growth suppression by down-regulation of cell surface growth factor receptors. PMID- 11894128 TI - Deficient R-smad/smad4 complex formation in fibroblasts growth-stimulated by TGF beta 1. AB - Although most cell types are inhibited by TGF-beta 1 some mesenchymal cells are growth-stimulated by TGF-beta 1. Here, using two cell models, in which the normal untransformed cells (WI38 human embryo and NIH3T3 mouse fibroblasts) are stimulated to proliferate by TGF-beta 1 and their SV40 Large T-transformed counterparts are inhibited, we show that the two former cells are deficient in smad2-smad4 and smad3-smad4 complex formation, whereas these complexes are present in the two latter cells. In addition, endogenous smad4 levels in total cell extracts are lower in WI38 and NIH3T3 fibroblasts than in their Large T expressing counterparts. In WI38 fibroblasts smad4 is, at best, inefficiently translocated to the nucleus. TGF-beta 1-mediated growth-stimulation of these normal fibroblasts does not appear to involve the presently known smads, but this situation is reversed in the Large T-transformed cells inhibited by TGF-beta 1. PMID- 11894129 TI - B7.1 immunogene therapy effectively activates CD(4+) tumor-infiltrating lymphocytes in the central nervous system in comparison with B7.2 gene therapy. AB - The B7 gene utilizing immunogene therapy is one of the most common methods against tumor growth. However, there is no known study that investigated the difference between B7.1 and B7.2 with regard to B7 gene therapy in the central nervous system (CNS). Therefore, to clarify the difference, we established B7.1 or B7.2 gene transduced tumor cells originating from the murine T cell lymphoma cell line EL4 (EL4-B7.1 or EL4-B7.2). First, we observed the survival time after intracranial inoculation of parent (IC-wt) or genetically modified tumor cells. All mice in control groups (IC-wt or IC-mock) were dead within 16 days. While there was significant survival elongation in the B7.2 modified group (IC-B7.2, p=0.0002), all mice in this group were dead of tumor growth within 22 days. On the other hand, 60% of mice inoculated with EL4-B7.1 (IC-B7.1) survived more than 120 days (p<0.0001). Second, to shed light on the anti-tumor immune response in situ, we tried to analyze CD(4+) tumor-infiltrating T lymphocytes (CD(4+) TIL). To purify and analyze CD(4+) TIL, we had to deplete F4/80(+) microglia because of the CD4 expression. In terms of activation marker expression in CD(4+) TIL, a small population was activated (CD25, 9.8%; CD69, 15.8%) in the control group (IC wt). In contrast, the activation marker positive CD4+ TIL percentage both in IC B7.1 (CD25, 25.1%; CD69, 40.1%) and IC-B7.2 (CD25, 16.2%; CD69, 28.3%) appeared to reflect the survival curve in both groups. These findings strongly suggest that, in the CNS, B7.1 gene therapy could effectively introduce CD(4+) TIL activation compared with B7.2 gene therapy. This is the first study clearly describing the difference between B7.1 gene therapy and B7.2 gene therapy in the CNS in terms of the activation status of CD(4+) TIL in situ. PMID- 11894130 TI - In vivo studies of the anti-tumor effects of a human prolactin antagonist, hPRL G129R. AB - Previously we demonstrated that a mutated human prolactin (hPRL) with a single amino acid substitution at position 129 (hPRL-G129R) was able to inhibit human breast cancer cell proliferation via the induction of apoptosis. In this study, we report the in vivo anti-tumor effects of hPRL-G129R in nude mice bearing human breast cancer xenografts (T-47D and MCF-7). In an effort to prolong the half-life of the proteins, hPRL or hPRL-G129R were formulated with either growth factor reduced Matrigel or into slow-releasing pellets (custom made 5 mg/5 day release). Initially, nude mice inoculated (s.c.) with T-47D human breast cancer cells were treated with either hPRL or hPRL-G129R formulated with Matrigel. At the end of the 7-week study, it was found that hPRL significantly stimulated the in vivo growth of T-47D xenografts (mean tumor volume, 202 +/- 62 mm(3) as compared to 124 +/- 31 mm(3) in control mice), whereas hPRL-G129R inhibited the tumor growth (mean tumor volume, 79+/-32 mm3). The inhibitory effects of hPRL-G129R were further confirmed in a second experiment using nude mice bearing MCF-7 human breast cancer xenografts and treated with slow-releasing pellets containing hPRL G129R. Based on these results, we believe that hPRL-G129R can be used to improve the outcome of human breast cancer treatment in the near future. PMID- 11894131 TI - Actinomycin D-mediated sensitization of AIDS-Kaposi's sarcoma cells to Fas mediated apoptosis: involvement of the mitochondrion-dependent pathway. AB - Fas engagement rapidly induces formation of the death-inducing signaling complex (DISC) that consists of Fas, FADD and pro-caspase-8. Activated caspase-8 at the DISC directly activates downstream caspases, resulting in induction of apoptosis of the independent mitochondria. In this study, we have obtained evidence demonstrating that Fas-mediated apoptosis in AIDS-KS cells takes place in a mitochondria-dependent manner. FADD and pro-caspase-8 were detected in immunoprecipitates with anti-Fas antibody in anti-Fas mAb (CH-11)-treated Hut 78, a typical Fas-sensitive cell line. On the other hand, DISC formation by CH-11 was markedly reduced in AIDS-KS cells. In addition, CH-11-induced activation of caspase-8-like protease in AIDS-KS cells was much less pronounced compared with that in Hut 78; however, a caspase-8 inhibitor, zIETD-fmk, completely blocked the apoptosis. Further, a caspase-9 inhibitor, zLEHD-fmk, markedly inhibited Fas mediated apoptosis in AIDS-KS cells. Several apoptotic stimuli induce mitochondria activation allowing cytochrome c release from the mitochondria. In the apoptosome, cytochrome c and Apaf-1 activate caspase-9 which subsequently leads to the activation of caspase-3. In AIDS-KS cells, CH-11 triggered cytochrome c release, an event which was inhibited by zIETD-fmk. Further, a caspase-3 inhibitor, zDEVD-fmk completely inhibited the apoptosis. Altogether, the present data provide evidence that the Fas signal in AIDS-KS cells is preferentially transduced through the mitochondria-dependent pathway, which is initiated by caspase-8 activation. PMID- 11894132 TI - Osteopenia in survivors of Wilms tumor. AB - Diminished bone mass (osteopenia) is recognized increasingly as a consequence of therapy in survivors of cancer in childhood. It has been reported in two small series of survivors of Wilms tumor. The objectives of this study were to explore, in a larger sample of such subjects, the prevalence of osteopenia and a possible relationship between osteopenia of the lumbar spine and abdominal irradiation. All survivors of Wilms tumor (n=49) in a single institution were considered eligible for study. Thirty-one agreed to participate; the non-participants were not notably different in their demographic characteristics and diseases/treatment experience. Information was obtained about prior treatment, and usual diet, sun exposure and physical activity. Bone mineral content was measured by dual energy X-ray absorptiometry, and biochemical markers of bone turnover, calciotropic hormones and minerals were assessed in a single blood sample. By Z-scores of whole body bone mineral content, 8 subjects were osteopenic. This was unrelated to milk intake or sun exposure and was not more common in the lumbar spine of those who had been irradiated (15/31 subjects). Physical activity correlated positively with bone mineral density Z-scores (p<0.005). Normal bone formation was reflected in normal blood levels of osteocalcin. C-telopeptide levels, reflecting bone resorption, were high but approximately correlated inversely with maturity. Low serum magnesium and parathyroid hormone levels were detected in a minority of subjects. Osteopenia is present in a large minority (27%) of survivors of Wilms tumor, and an imbalance of bone turnover (with excessive resorption) may be common. Irradiation does not appear to play a causal role. It is possible that a subtle renal tubular defect exists in these individuals; a prospect worthy of further exploration. PMID- 11894133 TI - Lack of NQO1 induction in human tumor cells is not due to changes in the promoter region of the gene. AB - DT-diaphorase is a two-electron reducing enzyme that is an important detoxifying enzyme and activator of bioreductive antitumor agents. Expression of the DT diaphorase gene, NQO1, appears to be transcriptionally regulated, and the gene is induced by a wide variety of compounds. We showed that 1,2-dithiole-3-thione can selectively increase DT-diaphorase activity in human and murine tumors, and that this enhanced the antitumor activity of bioreductive antitumor agents. However, we found that DT-diaphorase activity was not increased in some human tumor cell lines after treatment with the inducer, and this appeared to be due to a lack of increased transcription. To determine if this lack of increased transcription was due to a mutation in the promoter region of the NQO1 gene in these cells, we sequenced approximately 2000 bases of the NQO1 promoter region from non-induced cells and compared these with sequences from human HT29 colon cancer cells, which showed significant increases in NQO1 transcription, and the sequence reported for human liver cells in Genbank. Sequence analysis showed no changes in the sequences of the major transcriptional elements, XRE, CAT box, ARE, AP1 site or AP2 site, in the tumor cells compared with the Genbank sequences. The only major change was a deletion of a 20 base repeat region approximately 400 bases 5' to the XRE element in all the cells, including the HT29 cells, compared with the sequence reported in Genbank. There were also several insertions of a single base in various parts of the sequences which occurred in most, or all, of the cell lines compared with the reported sequence, and a small number of single base changes, insertions or deletions that occurred in a single cell line. However, these changes did not appear to correlate with differences in induced transcription of the NQO1 gene. These results suggest that the differences in transcription of the NQO1 gene after treatment with DT-diaphorase inducers was not due to alterations in the promoter region of the gene. PMID- 11894134 TI - A possible role for IVIg in the treatment of soft tissue sarcoma: a clinical case and an experimental model. AB - A patient with a malignant peripheral nerve sheath tumor (MPNST) was treated with IVIg for multiple sclerosis. Her MPNST course was remarkably longer and more indolent than expected; she achieved a disease-free interval (DFI) of 30 months. Seven other patients, who were not treated by IVIg, had a relatively aggressive course (median DFI 3 months). These results led us to examine the effect of IVIg on the growth of sarcoma in vitro and in vivo in an experimental model of MCA bearing mice. When added to MCA-105 sarcoma cell cultures, IVIg produced a dose dependent inhibitory effect on [(3)H]-thymidine incorporation. The maximal inhibitory effect was at a concentration of 50 mg/ml IVIg. Cell cycle analysis revealed a hypodiploid peak at the lower fluorescence values which appeared in the samples treated with IVIg. These results demonstrate that the anti proliferative activity results from an apoptotic effect of IVIg on the tumor cells. In a second set of experiments, we evaluated the capability of IVIg, when administered orally or subcutaneously, to inhibit the growth of MCA-105 sarcoma lung metastases. A decrease in the mean lung weight was observed in the mice that were treated by s.c. or oral administration, the latter being more effective. A possible role for IVIg in the treatment of MPNST and other soft tissue sarcomas is suggested. PMID- 11894135 TI - Dominant-negative activity of a Brca1 truncation mutant: effects on proliferation, tumorigenicity in vivo, and chemosensitivity in a mouse ovarian cancer cell line. AB - In order to generate an in vitro mouse model for the study of human ovarian cancers, we compared the effects of a truncated Brca1 mutant expression on cellular phenotype with those of a full-length sense and antisense Brca1 expression in the ID-8 mouse epithelial ovarian cancer cell line. The examined cellular processes include proliferation, tumorigenicity in syngeneic mice in vivo and sensitivity/resistance to several cytotoxic drugs. We found that the expression of a spontaneous truncated Brca1 mutant in ID-8 cells which contain two endogenous wild-type Brca1 alleles led to a dominant-negative effect of Brca1, demonstrated by an increase in tumorigenicity in vivo and in chemosensitivity. Expression of a truncated Brca1 mutant in a mouse epithelial ovarian cancer cell line could thus provide a powerful in vitro model for the study of human BRCA1-related ovarian tumorigenesis. PMID- 11894136 TI - Ceramide involvement in homocamptothecin- and camptothecin-induced cytotoxicity and apoptosis in colon HT29 cells. AB - Topoisomerase I inhibitors of the camptothecin (CPT) family have emerged as potent clinical chemotherapeutic agents in first-line treatment of solid colorectal cancer and in second-line for 5-fluorouracil resistant patients. CPT and homocamptothecin (hCPT), derivative with enhanced lactone stability, induced growth inhibition in HT29 cells via p53-independent apoptosis. hCPT- and CPT induced apoptosis was dependent on caspase-3 but not caspase-1. We report here substantial evidence that ceramide, resulting from de novo pathway or catabolism modulation, acted as a second messenger of these antitumor drugs in HT29 cells and leads to the activation of caspase-3. In addition, hCPT and CPT may favor ceramide signaling by disturbing sites of synthesis (Golgi) and trafficking of glucosylceramide from Golgi to lipid droplets. This work contributes to the understanding of the mechanism of action of CPTs, and suggests that inhibitors of glycosylation or activators of de novo metabolism could be of clinical interest in enhancing the effects of CPTs. PMID- 11894139 TI - JMM Past and Present. PMID- 11894137 TI - Lack of protein kinase C (PKC)-beta and low PKC-alpha, -delta, -epsilon, and zeta isozyme levels in proliferating human melanoma cells. AB - Protein kinase C (PKC), a calcium and phospholipid-dependent kinase, has been implicated in carcinogenesis of melanocytic cells. However, its role in melanoma cell growth remains controversial. We therefore investigated the growth dependence of PKC isozyme expression in human normal melanocytes and melanoma cells. Logarithmic and stationary growth phases in culture were clearly distinguished by nuclear cell staining with the proliferation marker Ki-67. PKC beta I and -beta II were expressed exclusively in normal melanocytes but not in melanoma cells, whereas PKC-gamma was not found in any of the cultures studied. Low PKC-delta, -epsilon and -zeta mRNA levels were detected by RT-PCR in proliferating melanoma cells and higher in confluent non-proliferating cells, whereas levels of PKC-alpha mRNA remained rather stable. Subcellular fractionation and immunoblotting revealed accordingly low expression of PKC alpha, -delta, -epsilon, and -zeta in the logarithmic growth phase of melanoma cells, with subsequent increase of expression and of membrane association in the stationary phase. Only weak differences were detected between the growth phases in normal melanocytes for the respective PKC isozymes, except for membrane associated PKC-beta I and -beta II which were clearly elevated in confluent melanocyte cultures. These data suggest that certain PKC isozymes are involved in the intracellular signalling that regulates melanoma cell proliferation, and may function as suppressors of tumour cell growth. PMID- 11894138 TI - Estrogen and atherosclerosis. PMID- 11894140 TI - Apoptosis and chemoresistance in transgenic cancer models. AB - Multidrug resistance remains an unresolved problem in clinical oncology. Over a decade ago genes encoding cellular efflux pumps were shown to confer resistance to a broad spectrum of biochemically unrelated anticancer drugs even before the compounds reached their intracellular targets. More recently it has become apparent that many drugs induce a common apoptotic program, such that mutations in this program can also produce multidrug resistance. However, a thorough evaluation of the contribution of apoptotic defects to this "postdamage" drug resistant phenotype is technically complicated, and this has led to uncertainty about the overall significance of apoptosis in therapy-induced cell death. For example, correlative analyses using patient specimens are limited by unknown background mutations in the biopsy material, and assays using cancer cell lines can be biased by unphysiological conditions. We sought to circumvent these restrictions by utilizing a tractable transgenic cancer model to examine the impact of apoptosis on treatment outcome. Here we discuss potential caveats of cell culture based assays, highlight features of genetically engineered mice as potential model systems, and describe a tractable transgenic mouse model to study drug responses in a series of primary lymphomas with genetically defined lesions treated at their natural site. PMID- 11894141 TI - Immunoregulatory functions of interleukin 18 and its role in defense against bacterial pathogens. AB - Interleukin-18 (IL-18) is a proinflammatory cytokine that belongs to the IL-1 cytokine family, due to its structure, receptor family and signal transduction pathways. Similarly to IL-1beta, IL-18 is synthesized as a precursor requiring caspase-1 for cleavage into an active IL-18 molecule. However, with regard to its capacity to induce the production of Th1 cytokines and to enhance cell-mediated cytotoxicity, IL-18 is also related to IL-12. Produced mainly by antigen presenting cells, IL-18 is a pleiotropic factor involved in the regulation of both innate and acquired immune responses, playing a key role in autoimmune, inflammatory, and infectious diseases. This review summarizes recent advances in the understanding of IL-18 structure, processing, receptor expression, and immunoregulatory functions and emphasizes the critical role of this cytokine in bacterial infections. It focuses on the participation of this cytokine in the defense against intracellular bacteria, including Listeria, Shigella, Salmonella, and Mycobacterium tuberculosis. Since this cytokine may be particularly useful in immunoprophylactic and immunotherapeutic interventions in which the cellular response is most desirable, the potential therapeutic aspects of IL-18 is also discussed. PMID- 11894142 TI - Replication-selective viruses for cancer therapy. AB - Advances in our understanding of the molecular basis of cancer and the availability of technology to genetically engineer viruses have led to the development of replication-competent viruses to treat cancer. In theory, replication-selective viruses offer several appealing properties as biological agents for cancer therapy: they kill tumor cells selectively, and their replication leads to amplification of their oncolytic potential. Most preclinical experiments in tissue culture and in animal models support this notion. Clinical data on the first generation of replication-selective viruses are now rapidly accruing. The therapeutic index, and ultimately the clinical outcome, will depend on a complex balance between host and viral factors. This review discusses strategies to kill cancer cells based on our understanding of their molecular defects and the progress being made using replication-competent viruses for tumor therapy. We focus our discussion on a replication-selective adenovirus called ONYX-015 that has recently demonstrated encouraging results in clinical trials PMID- 11894143 TI - Coronary artery wall atherosclerosis in relation to the estrogen receptor 1 gene polymorphism: an autopsy study. AB - Estrogen receptors (ESR) 1 and 2 are expressed in the normal and atherosclerotic arteries mediating the atheroprotective action of estrogen to artery wall cells. Whether variants of these receptor genes associate with autopsy-verified coronary artery wall atherosclerosis is not known. This study investigated whether variants of the ESR1 gene are associated with autopsy-verified coronary artery wall atherosclerosis and thrombosis. Coronary arteries were taken from 300 white Finnish male autopsy cases aged 33-69 years included in the Helsinki Sudden Death Study. Areas of coronary wall covered with fatty streaks, fibrotic, calcified, and complicated lesions were measured using computer-assisted planimetry and related to ESR1 PvuII genotypes (P/P, P/p, and p/p) determined by PCR. The mean area of complicated lesions of three major coronaries and the presence of coronary thrombosis were significantly associated with the ESR1 genotype in men aged 53 years or older (median age as a cut off point). No such association was found in men aged under 53 years. After adjusting for age and body mass index the men aged 53 years or over with P/p and P/P genotype had areas of complicated lesions on average two- and fivefold larger than subjects with the p/p genotype. The age and body mass index adjusted odds ratios for coronary thrombosis were 6.2 for P/p and 10.6 for P/P compared to men with the p/p genotype. After additional adjustment for diabetes and hypertension the ESR1 genotype persisted as an independent predictor of complicated lesions ( P=0.007) and coronary thrombosis. In conclusion, the ESR1 gene is a potential candidate behind the pathogenesis of acute coronary events. PMID- 11894144 TI - Effect of ecNOS polymorphisms and coronary artery disease upon exhaled nitric oxide. AB - Exhaled nitric oxide (eNO) is thought to arise principally from the airway epithelium. NO regulates smooth muscle tone, and abnormal activity of NO synthase has been implicated in coronary artery disease (CAD). Polymorphisms of endothelial constitutive NO synthase (ecNOS) may affect NO generation and be associated with CAD. It was hypothesised that a polymorphism, such as the ecNOS intron 4 polymorphism (ecNOS4a), affects the levels of eNO via airway epithelial NOS. eNO levels were measured in 53 patients with ischaemic chest pain who had previously been genotyped for ecNOS polymorphisms, with sample enrichment for the ecNOS4a allele. Subjects were also assessed for two other ecNOS polymorphisms (T 786C substitution in the promoter region, and G5557T in exon 7), variably associated with vascular disease. Those homozygous for the 'a' allele (ecNOS4a/a) had a lower mean eNO (9.0 ppb) than those who were heterozygous (ecNOS4a/b, 13.6 ppb), who in turn had a lower level than those homozygous for the wild-type ecNOS4b/b (16.1 ppb). No association of eNO levels was found with the other polymorphisms. Levels of eNO remained significantly lower in the ecNOSa/a subjects than in the ecNOSa/b and ecNOSb/b subjects, even when controlled for angiographic CAD, and smoking habit. In addition, all subjects with CAD had a significantly lower mean eNO (12.1 ppb) than subjects without angiographic CAD (19.9 ppb). In this selected population low levels of eNO were thus associated with presence of the ecNOS4a allele and also with CAD. PMID- 11894145 TI - Inhibition of duck hepatitis B virus replication by intrahepatic expression of an antiviral resistance gene. AB - Resistance genes coding for inhibitors of hepadnaviral replication, such as ribozymes, antisense RNA, and dominant negative mutants have been shown to be effective in transfected hepatoma cells. In vivo studies, however, are not available to date. Here we expanded the use of the duck hepatitis B virus (DHBV) model for studying antiviral resistance genes in vivo. Animals were experimentally infected by intravenous injection of DHBV-positive serum in ovo. The use of recombinant human adenovirus type 5 and avian adenovirus CELO for gene transfer was evaluated. Adenovirus type 5 transduced more than 95% and CELO less than 1% of embryonic hepatocytes in vivo. Adenovirus type 5 interfered with DHBV replication (viral cross-talk), but this effect was moderate and did not preclude analysis of specific antiviral effects. Thus adenoviral transfer of a dominant negative mutant prior to DHBV infection (intracellular immunization) yielded 100 fold suppression of viral replication compared to the green fluorescent protein marker gene. Neither gene was toxic. These data demonstrate that a prototype anthepadnaviral resistance gene is functional in vivo. Duck embryos represent a useful model for evaluating gene therapeutic strategies in vivo without the need for large scale preparations of gene delivery vehicles. PMID- 11894146 TI - Pediatric cardiologists: counselors versus enforcers. PMID- 11894149 TI - Frequency-domain analysis of the QRS complex after treatment of childhood cancer with anthracycline cytostatics. AB - Long-term cardiac complications, occurring several years after completion of anticancer treatment, may develop from subclinical myocardial damage induced during cardiotoxic therapy. The aim of this study was to evaluate the usefulness of frequency-domain signal-averaged ECG analysis of the QRS complex for assessing the cardiotoxicity of anthracycline cytostatics. Altogether, 172 signal-averaged electrocardiography (SAECG) registrations were performed in 50 repeatedly evaluated oncologic patients. These registrations were performed 0.2-15 years after completion of anthracycline therapy for childhood cancer. The control group consisted of 120 healthy children and young volunteers; in 20 of these controls, SAECGs were performed repeatedly. Using gliding window fast Fourier transformation within the QRS complex, values area ratio (AR) 60-120 Hz/0-120 Hz were calculated in X, Y, and Z lead. Area ratio of patients after anthracycline therapy was significantly higher than those in control group in X lead. Differences in frequency content in the QRS complex between patients and controls might signal an initial stage of anthracycline-induced myocardial damage. PMID- 11894147 TI - Myocardial performance and baroreceptor reflexes in preterm neonates: an echocardiographic evaluation using the tilt-table test. AB - The hemodynamic consequence of head-up position in preterm infants is not known, so we used the tilt-table test to assess changes in myocardial performance and baroreceptor reflexes. Twenty-five preterm infants with gestational age (GA) (mean +/- SD) 31 +/- 2.9 weeks (range 25-35.5 weeks), birth weight of 1612 +/- 642 g (range 520-3260 g) were studied in supine, 30 degrees, and 60 degrees head up positions. GA had a significant effect on heart rate (HR) (p = 0.007), systolic blood pressure (SBP) (p = 0.03), left ventricular (LV) cavity dimensions (p = 0.001), cardiac output (CO) (p = 0.001), LV ejection time (LVETc) (p = 0.05), and end systolic wall stress (ESWS) (p = 0.003). An inverse relationship was seen between velocity of circumferential fiber shortening (VcFs) and ESWS (slope b = -0.019 +/- 0.008, p = 0.003). Results of tilt tests showed that at supine, 30 degrees, and 60 degrees, respectively, HR was 162 +/- 10.5, 162 +/- 9, and 164 +/-12 (p = NS); SBP (mmHg) was 73 +/- 11, 72.5 +/- 9.5, and 78 +/- 10 (p = NS); CO (L/kg/min) was 0.4 +/- 0.16, 0.4 +/- 0.15, and 0.43 +/- 0.16 (p = NS); ESWS (g/cm2) was 38.7 +/- 8.3, 40.9 +/- 9.9, and 43.4 +/- 10.7 (p = NS); and VcFs (circ/sec) was 1.35 +/- 0.3, 1.28 +/- 0.4, and 1.26 +/- 0.2 (p = NS). LV filling pattern as seen by early/late atrial Doppler flow velocity ratio did not change with tilt (p = NS). Myocardial performance improved with increasing GA. No significant differences in myocardial performance were found between baseline and head-up tilt positions. PMID- 11894148 TI - Tracheal compression by elongated aortic arch in patients with congenitally corrected transposition of the great arteries. AB - We investigate the tracheal compression by aortic arch in patients with congenitally corrected transposition of the great arteries. Fourteen patients whose ascending aorta was connected to descending aorta on the contralateral side were divided into two groups according to the severity of tracheal compression on lateral angiogram: group 1 (stenosis > or = 50%, eight patients) and group 2 (stenosis < 50%, six patients). We compared the following variables between the groups: aortic size, lateral displacement of ascending aorta, retrosternal space, and contour of aortic arch. Spiral computed tomography (CT) was done in five patients of group 1. The ratio between measured and normal diameters of ascending aorta of group 1 was significantly larger than that of group 2 (1.57 +/- 0.14 compared to 1.19 +/- 0.26, p = 0.014). There was linear correlation between the tracheal compression and aortic size (r = 0.69, p = 0.001). The ascending aorta was positioned more laterally and the aortic arch was located more posteriorly in group 1. In four patients with decreased tracheal compression after surgery, aortic size decreased and the ascending aorta restored its anterior position. Spiral CT showed transversely oriented aortic arch and severe tracheal compression in the anteroposterior direction by aortic arch. The tracheal compression by aortic arch in corrected transposition is common when the descending aorta is located on the contralateral side of the ascending aorta. Aortic size, posterior position, elongation, and end-on appearance of the aortic arch are the useful predictors of tracheal compression. PMID- 11894150 TI - Second-line treatment of fetal supraventricular tachycardia using flecainide acetate. AB - Digoxin has been an effective treatment for fetal supraventricular tachycardia (SVT), but second-line therapy remains more controversial. Thirty-seven cases of fetal SVT were identified that received digoxin as first-line therapy. Seventeen fetuses (46%) converted to and maintained normal sinus rhythm. Flecainide was used in 13/15 patients requiring second-line therapy; 12/13 (92%) converted to sinus rhythm. Of seven hydropic fetuses, five required second-line therapy and were then successfully converted with flecainide. The improved efficacy of flecainide was statistically significant with a p value <0.01. Complete follow-up was available in 13 digoxin-treated and in 12 second-line therapy infants. Prolonged or multiple drug therapy for postnatal arrhythmia management was required in 3/13 (23%) patients in the digoxin group and in 8/12 (67%) patients requiring second-line therapy. This demonstrated a correlation between the need for second-line fetal therapy and more complex postnatal management with a p value of 0.003. Digoxin remains an effective first-line therapy in the treatment of fetal SVT. Flecainide is an effective second-line therapy, especially in the face of fetal hydrops. Use of second-line therapy in fetal SVT is a predictor of complex postnatal course, and these patients should be followed more closely. PMID- 11894151 TI - Propofol does not modify the hemodynamic status of children with intracardiac shunts undergoing cardiac catheterization. AB - Immobility and cardiovascular stability are required for cardiac catheterization. Pediatric patients need a type of sedation that also allows spontaneous ventilation without supplemental oxygen. Propofol has been adequate in hemodynamically stable patients with congenital heart disease undergoing cardiac catheterization. However, mild systemic hypotension caused by propofol may increase a preexisting right-to-left shunt. The aim of this study is to evaluate, in pediatric patients scheduled for cardiac catheterization, the effects of propofol on systemic and pulmonic circulations. Fifteen patients aged 18 months to 9 years were studied. After a fast of 4-6 hours for solid food, the patient arrived at the cardiac catheterization suite, where an IV catheter was placed. Usual monitoring was used. For sedation, without supplemental oxygen, patients received 1 mg/kg of fentanyl followed by propofol (1-2 mg/kg) titrated to immobility during preparation of the groin. A continuous infusion of propofol (100 mg/kg/min) was also started to obtain immobility during the procedure. Hemodynamic data, including systemic venous, pulmonary artery and vein, aortic saturations, and pressures, were recorded; Qp and Qs were calculated. The same set of data was re-corded 4 minutes after discontinuation of propofol and when the patient was responding to tactile stimuli. Despite lower pressures during propofol infusion, as compared with those pressures measured after discontinuation of propofol, the extent of the intracardiac shunt remained unchanged. Propofol seems to be an adequate sedative agent for pediatric patients undergoing cardiac catheterization, including those with intracardiac shunts. PMID- 11894152 TI - Increase in diameter of ventricular septal defect and membranous septal aneurysm formation during the infantile period. AB - Serial changes in diameters of ventricular septal defect (VSD), and in the formation patterns of membranous septal aneurysm (MSA), were evaluated using two dimensional (2-D) echocardiography during the infantile period. We studied 65 patients with VSD, who were classified into three groups by clinical feature. The patients with severe respiratory symptoms and significant failure to thrive were classified as group A (n = 15). The patients with no respiratory symptoms and mild or moderate failure to thrive were classified as group B (n = 38). Group C had no or trivial clinical symptoms and no need for surgical repair (n = 12). Group A showed rapid enlargement of VSD during early infancy. In this group, the mean diameter of VSD at age 5 months was 75% larger than the mean diameter at age 1 month (an increase of 5.5 to 9.6 mm; p<0.05). The other two groups (B, C) showed a slower enlargement as compared with group A. The rapid enlargement of VSD in group A seemed to induce the rapid increase in pulmonary blood flow with severe clinical symptoms. We suggest that careful observation of the changes in VSD diameters by periodic echocardiography is important in predicting the clinical course and in deciding the operation indication. PMID- 11894153 TI - Signal-averaged electrocardiography in children with anthracycline-induced cardiomyopathy. AB - The aim of the present study was to determine if signal-averaged ECG of patients with anthracycline-induced left ventricular dysfunction could differentiate between patients with anthracycline-induced cardiotoxicity and those without. Sixteen children with anthracycline-induced cardiomyopathy, aged 6.5 to 15.5 years (anthracycline dose = 198-737 mg/m2), and 31 patients aged 5.0 to 16.7 years, who received anthracyclines without evidence of left ventricular dysfunction (anthracycline dose = 120-517 mg/m2), were studied with signal averaged ECG. The two groups were comparable in age, body surface area, and time since completion of chemotherapy. Signal averaged ECG parameters of the patients were compared with data obtained from 530 healthy children. These parameters were converted to z-scores to account for growth-related changes in signal averaged ECG recordings. Z-scores for filtered QRS duration and low amplitude terminal signal < 40 microV were significantly lower (p = 0.002 and p = 0.015, respectively), and Z-score for root mean square voltage of the last -30 ms of filtered QRS tended to be higher (p = 0.06) in patients with left ventricular dysfunction. Filtered QRS duration lower than -1.5 SD was found in 4 of 16 patients with left ventricular dysfunction and in only 1 of 31 patients without (p < 0.05) yielding a sensitivity of 25% and a specificity of 97% to detect left ventricular dysfunction. Only 1 patient had late potentials; his left ventricular function was normal. Left ventricular mass index tended to be lower in patients with left ventricular dysfunction (p = 0.07), whereas left ventricular diastolic diameter was similar in the two groups. The mechanism that accounted for the difference in signal averaged-ECG between the two groups of patients could be linked with the decrease in left ventricular mass in patients with left ventricular dysfunction. In conclusion, children with left ventricular dysfunction following anthracycline therapy have a SA ECG different from those without left ventricular dysfunction, which is mainly characterized by a lower filtered QRS duration. A prospective study is needed in order to determine if this modification of SA ECG recordings precedes alteration of left ventricular function, and, therefore, if it could help in early detection of cardiac toxicity of anthracyclines. PMID- 11894154 TI - Left ventricular functions in children with chronic pulmonary diseases. AB - We evaluated left ventricular (LV) functions in 32 patients with chronic pulmonary diseases (CPDs) and compared them to 16 healthy controls. Echocardiographic assessment of both systolic and diastolic parameters of LV was performed for all patients and controls. LV systolic functions were found to be preserved in mild states, but with evident hypoxia the aortic acceleration and deceleration times decreased. LV diastolic functions were impaired in cases with pulmonary hypertension as reflected by a decreased ratio of peak flow velocity of early filling to peak flow velocity of late filling. PMID- 11894155 TI - Flow pattern of the superior caval vein in children after closure of atrial septal defect: a comparison of catheter therapy with open-heart surgery. AB - To compare the flow pattern of the superior caval vein (superior vena cava; SVC) after catheter therapy with that after open-heart surgery, we examined the flow velocity curves of the SVC in 10 pediatric patients who underwent catheter closure of atrial septal defect (ASD) and in 20 pediatric patients who underwent open-heart surgery. We used pulsed Doppler examinations to record the velocity curves. On the velocity curve of the SVC in children after open-heart surgery, the velocity in systole was lower than that in diastole, which is contrary to the normal pattern. The velocity in systole increased more than that in diastole during inspiration, which also differs from the normal pattern. In contrast, the velocity curve of the SVC in patients after catheter therapy showed a normal pattern. The change of flow velocity during respiration became smaller after treatment in the surgery group, although it did not change after treatment in the catheter group. These results might indicate less reserve ability in the right atrium in patients who undergo open-heart surgery. ASD closure by catheter is useful not only for its economy and noninvasiveness but also for its potential to contribute conservation of right atrial function. PMID- 11894156 TI - Exercise testing and 24-hour ambulatory blood pressure monitoring in children with Williams syndrome. AB - The aim of the study was to assess workload capacity and blood pressure (BP) response to treadmill exercise and 24-hour BP monitoring in children with Williams syndrome. Seventeen children were examined (8 males and 9 females) whose mean age was 13.8 +/- 3.6 years. Six patients were on antihypertensive therapy. Each patient underwent clinical examination and measurement of BP at rest, during exercise, and during 24-hour monitoring. Two-dimensional echocardiogram and echo Doppler of renal arteries were performed. The test was stopped for muscular fatigue or reduced cooperation. The patients, when compared to a population of healthy children, had reduced total time of exercise (7.3 +/- 1.9 vs 14.3 +/- 2.6 min, p < 0.001) and, at the same workload, increased heart rate (167 +/- 19 vs 145 +/- 16 beats/min, p < 0.001) and increased maximum systolic BP (146 +/- 27 vs 128 +/- 12 mmHg, p = 0.01). Ambulatory blood pressure measurement values showed higher systolic blood pressure both during daytime and nighttime. Our study confirms that children and adolescents with Williams syndrome are at high risk for hypertension, probably related to the alterations of large arteries. The data relating to the synthesis of elastin may have a direct relationship to the compliance of the arterial system, leading to hypertension. PMID- 11894157 TI - Isolated noncompaction of ventricular myocardium associated with fatal ventricular fibrillation. AB - A female infant with isolated noncompaction of ventricular myocardium who developed ventricular tachyarrhythmia is described. Wolff-Parkinson-White syndrome was shown by electrocardiography. At 9 months of age, the patient suddenly developed cardiac arrest. Electrocardiography following resuscitation with DC cardioversion demonstrated sinus rhythm without delta wave. The QT interval was normal. Frequent premature ventricular captures caused ventricular fibrillation. DC cardioversion was necessary to terminate frequent attacks of ventricular fibrillation until the introduction of beta blockers and lidocaine. Two-dimensional echocardiogram confirmed the diagnosis of isolated non-compaction of ventricular myocardium. Three months later, the patient died of ventricular fibrillation during respiratory syncytial viral infection. PMID- 11894158 TI - Around PediHeart; prenatal counseling. PMID- 11894160 TI - Balloon valvuloplasty for critical pulmonary valve stenosis in a premature infant. AB - Successful balloon valvuloplasty for critical pulmonary valve stenosis is described in an 800-g infant. A modified catheter was required to cross the valve in the smallest child known to undergo this procedure. PMID- 11894159 TI - Rhabdomyoma in the fetus: illustration of tumor growth during the second half of gestation. AB - Rhabdomyomata are the most common cardiac tumors in childhood and are associated with tuberous sclerosis. These tumors tend to regress in the first years of life. Little is known about their intrauterine growth pattern. We describe three fetuses with cardiac rhabdomyomata and illustrate the tumor growth by serial echocardiographic views during the second half of gestation. Tumor growth is proportional to cardiac growth and tends to be somewhat slower toward the end of pregnancy. PMID- 11894161 TI - Pseudoaneurysm of the right ventricular outflow tract complicating balloon dilatation for tetralogy of fallot. AB - Balloon dilatation is one of the treatment options in symptomatic infants with tetralogy of Fallot and hypoplastic pulmonary annulus and pulmonary artery. A balloon dilatation was performed on a 28-day-old infant with tetralogy of Fallot with an appropriate balloon. The patient developed two pseudoaneurysms on the right ventricular outflow tract after the procedure which were diagnosed when the patient was admitted for total correction at 20 months of age. This case demonstrates an unusual but potentially life-threatening long-term complication of this procedure. PMID- 11894163 TI - Balloon pericardiotomy for recurrent pericardial effusions following fontan revision. AB - Percutaneous balloon pericardiotomy (PBP) has been used in patients with recurrent pericardial effusions in which the chest and pericardium had not been previously entered and therefore concerns about adhesions were minimal. Few reports have depicted PBP for patients with recurrent pericardial effusions following open-heart surgery, in which adhesions may play a greater role. We report a successful creation of a pericardial window using PBP in a child with recurrent pericardial effusions after a Fontan revision. This technique offers a simpler approach to the treatment of recurrent pericardial effusions after open heart surgery in patients with 8 congenital heart disease. PMID- 11894162 TI - Functional pulmonary atresia in neonatal Marfan's syndrome: successful treatment with inhaled nitric oxide. AB - Functional pulmonary atresia is characterized by a structurally normal pulmonary valve not opening during right ventricular ejection. We report this rare condition in a premature newborn of a twin pregnancy, in which fetal echocardiography findings were consistent with critical pulmonary stenosis. After birth, features of neonatal Marfan's syndrome were noted. Echocardiography showed a morphologically normal but immobile pulmonary valve with continuous regurgitation. Right ventricular pressure was subsystemic. In this case, initial treatment with nitric oxide, followed by pharmacological duct closure, was successful. Differentiating between anatomic and functional pulmonary valve atresia may be difficult. The echocardiographic criteria are discussed. PMID- 11894164 TI - Fatal pulmonary hypertension associated with an atypical case of Hunchinson Gilford progeria. AB - We report the first autopsied case of an incomplete type of Hunchinson-Gilford progeria associated with fatal pulmonary hypertension. Histopathologic findings revealed abnormal deposition of collagen and elastic fibers, as well as cellular proliferation, at intima of the coronary arteries, pulmonary small arteries, and arterioles. These changes could be underlying conditions of the association of the two diseases. PMID- 11894165 TI - Around PediHeart: prenatal counseling--part two. PMID- 11894166 TI - Quantitation of localized abnormal deformation in asymmetric nonobstructive hypertrophic cardiomyopathy: a velocity, strain rate, and strain Doppler myocardial imaging study. AB - We report a case of a 10-year-old child with nonobstructive hypertrophic cardiomyopathy in whom two-dimensional echocardiography showed asymmetric septal hypertrophy with a localized thickening in the mid-septal segment. Systolic regional longitudinal motion and deformation indices were quantified by the new ultrasound-based parameters velocity, strain rate, and strain. Regional longitudinal myocardial function indices were normal for the basal and apical septal segments. The deformation parameters strain rate and strain (not the regional velocity profile) were abnormal only in the hypertrophied mid-septal segment with myofibril disarray. The concepts and advantages and clinical implications behind this quantitative approach to localizing and quantifying areas of abnormal deformation related to such myocardial disarray in localized hypertrophy are discussed. PMID- 11894172 TI - Abstracts of the 46th Annual meeting of the GTH (Gesellschaft fur Thrombose- und Hamostaseforschung) February 20-23, 2002, Erfurt. PMID- 11894167 TI - Severe pulmonary hypertensive vascular disease in two newborns with aneurysmal vein of galen. AB - Arteriovenous malformation of the vein of Galen (AVG) is a rare entity in the newborn with a high morbidity and mortality. We present two cases of fatal AVG with persistent pulmonary hypertension of the newborn and significant pulmonary hypertension documented by autopsy histopathology. The pathophysiology is reviewed and a proposed mechanism of the association between AVG and pulmonary hypertension is discussed. PMID- 11894174 TI - [Percutaneous revascularization of multivessel coronary disease using stents - a multicenter, prospective study]. AB - BACKGROUND AND OBJECTIVE: Symptomatic patients with multivessel coronary disease (MVD) benefit from both coronary artery bypass grafting (CABG) and percutaneous coronary angioplasty (PTCA). The >>German Angioplasty Bypass Investigation<< (GABI-I) trial randomized patients to one of these treatment strategies between 1986 and 1991. In order to evaluate the impact of current technology, in particular coronary stents, the GABI-II trial was initiated, which in 1996 and 1997 prospectively enrolled patients according to the initial GABI-I criteria. PATIENTS AND METHODS: Into the study 136 consecutive patients (108 men, 28 women; 63 +/- 12 years) were included. Patients from GABI-I served as controls. RESULTS: A mean of 2.1 +/- 0.5 vessels were treated per patient (vs. 1.9 +/- 0.5 vessels in the PTCA arm of GABI-I) and 63 % of the lesions were covered with stents. With respect to the primary endpoint less patients remained with a CCS class III or IV in GABI-II after 12 months (1,5 % vs. 8 % in the PTCA arm of GABI-I, p<0,01). No patient required emergency or urgent bypass operation in GABI-II (vs. 9 % in GABI I, p < 0.01). After 12 months, 8 % of the patients were sent for bypass surgery (CABG) vs. 21 % in GABI-I (p < 0.001), and 20 % (vs. 23 % in GABI-I) of the patients underwent Re-PTCA. The percentage of patients without reinterventions was 72 % vs. 56 % in GABI-I (p < 0.01), but remained lower compared to patients randomized to CABG in GABI-I (94 %, p < 0.001). CONCLUSION: PTCA in patients with MVD is still associated with a higher reintervention rate as compared with CABG. However, in contrast to angioplasty a decade ago, PTCA in conjunction with stents significantly lowered the need for subsequent revascularization, which was mainly driven by the reduced necessity for bypass surgery. PMID- 11894175 TI - [Umbilical metastasis of a gallbladder carcinoma: "Sister Mary Joseph's nodule"]. AB - HISTOLOGY AND CLINICAL FINDINGS: A 63-year-old man presented with a firm and crusting umbilical mass which had developed within several weeks. The tumor was 1.5 cm in diameter. There were no other diseases in the patient's history. Physical examination was normal except the umbilical tumor. INVESTIGATIONS: Histologic examination revealed a malignant epithelial tumor corresponding to a metastasis of an adenocarcinoma which could be a carcinoma of the gallbladder or the pancreas. Initially the following clinical examination of the patient showed no pathological findings: gastroscopy and coloscopy gave no hint for a primary carcinoma. Computed tomography of the abdomen revealed a thickened gallbladder wall with an irregular intraluminal contour suspicious of a gallbladder carcinoma. During surgery the gallbladder showed a clearly thickened wall. There were numerous carcinomatous lesions on the peritoneum. A cholecystectomy was performed and the umbilical metastasis was excised. DIAGNOSIS, TREATMENT AND COURSE: Histologic examination revealed a metastatic adenocarcinoma of the gallbladder with tumorous infiltration of the liver tissue and a solitary metastasis in the skin (>>Sister Mary Joseph's nodule<<). During wound healing there were no complications and the patient was demitted 10 days after surgery. Following a recovery phase at home a palliative chemotherapy was planned. Because the patient's general condition worsened he was readmitted to the hospital. He presented with a considerable anemia and an icterus. Sonography revealed a hilar tumor measuring 4 cm in diameter. The intrahepatic bile ducts were congested. A stent was inserted by endoscopic retrograde pancreaticocholangiography. During the following weeks the patient's general condition worsened rapidly and he died 9 months after the skin metastasis had occurred. CONCLUSION: A rapidly developing umbilical mass (>>Sister Mary Joseph's nodule<<) is suspicious of an underlying metastatic adenocarcinoma. The necessary examination and therapy including excision of the umbilical metastasis is warranted without delay. PMID- 11894176 TI - [Puumala virus infection (nephropathia epidemica) as different diagnosis of acute renal failure]. AB - Puumala virus infection (nephropathia epidemica) as different diagnosis of acute renal failure. HISTORY: A 34-year old patient presented in reduced status with a sudden onset of fever, headache, backpain, abdominal pain, mild diarrhea, nausea with vomiting, and blurred vision. Within a few days an acute renal failure developed. INVESTIGATIONS: On admittance there was thrombocytopenia of 27/nl, CRP of 109 mg/l and proteinuria of 5 g/l, moderate glucosuria and erythrocyturia of 250/microliter. Renal biopsy showed acute hemorrhagic interstitial nephritis. DIAGNOSIS, THERAPY AND FOLLOW UP: Diagnosis of nephropathia epidemica was proven by puumala-virus IgM- and later IgG-antibodies. Hantaan-antibodies were negative. Maximum serum creatinine of 640 micromol/l and urea of 30.5 mmol/l developed on the 5(th) day after admission. Without specific therapy the patient recovered fast and there were no persisting abnormalities during a 2-year follow up. CONCLUSION: In young patients with acute renal failure of unknown origin with the above symptoms hantavirus-infection with the subtypes puumala and dobrava should be considered in Central Europe. PMID- 11894178 TI - [Sonographic diagnosis of thrombosis]. PMID- 11894177 TI - [Treatment of chronic hepatitis C with pegylated alpha-interferons]. PMID- 11894179 TI - [Forced labour and public health under national socialism: specialized literature and research perspectives]. PMID- 11894180 TI - [How to treat hyperuricemia and gout attacks in compensated kidney failure?]. PMID- 11894181 TI - [Tumor marker determination errors]. PMID- 11894182 TI - [What measures are necessary in the demonstration of Clostridium difficile toxin?]. PMID- 11894183 TI - [Pneumonia diagnosis in the practice on the test-bench: how much diagnosis is necessary?]. PMID- 11894184 TI - [The dilemma of violence in psychiatry]. PMID- 11894185 TI - [Violent behaviour of patients in institutions]. AB - OBJECTIVE: The assessment of violence risk has impact on psychiatrists' decisions not only in forensic psychiatry but concerns many aspects of civil psychiatric units, too. Within the last decade, many studies on violence have been published. The results are misinterpreted sometimes. METHOD: A review about studies on in patient violence is given. RESULTS: In-patient violence and violence in the community should strictly be separated concerning predictors and predictions. Predictors of community violence both for persons with major mental disorders and persons without mental illness are criminal history, male gender, younger age, and substance abuse. Psychopathological and clinical variables play a minor role. In contrast, in-patient violence is closely related to the severity of psychopathological symptoms. More detailed determinations of risk-related symptoms yield conflicting results due to inevitable problems of sample selection. CONCLUSIONS: The assessment of violence risk has to consider the different risk constellations for violence in institutions and in the community. Predictions of violence by individual risk variables are limited because situation-related factors are important, too. PMID- 11894186 TI - [Employment situation of psychiatric in-patients with schizophrenic or affective disorders]. AB - OBJECTIVE: This study is on employment on the competitive as well as the sheltered work market of psychiatric in-patients suffering from schizophrenia or affective disorders. Furthermore it investigates the importance of characteristics of disorder and of educational and vocational background as predictors of occupation. METHODS: A sample of 251 in-patients participated in a structured interview during their hospitalization. RESULTS: 43 % of in-patients suffering from schizophrenia and 46 % suffering from affective disorders were employed on the competitive work market during the year before index hospitalization. Many were without any occupation too because sheltered employment was rarely used and mostly by schizophrenics (17 % vs 4 % of persons with affective disorders). Predictors of employment are past vocational experiences, education, length of past hospitalization, psychiatric diagnosis and sex. CONCLUSIONS: Results raise the question of deficits in providing with sheltered employment opportunities for the mentally ill and in particular for people suffering from affective disorders. Alternatives to these deficits are discussed. PMID- 11894187 TI - [Difficulties in the evaluation of depressive symptoms in patients with dementia]. AB - Depressive symptoms are often found in dementia (up to 86 %). Therefore, adequate treatment is necessary. Depressive symptoms appear significantly more often in vascular dementia than in dementia in Alzheimer's disease, but severity and profile of depressive symptomatology are independent of the etiology of dementia. The aim of this review is to help clinicians to select appropriate psychometric instruments to identify and measure depressive symptomatology in dementia. Frequently used scales are described, e.g. the Dementia Mood Assessment Scale and the Cornell Scale for Depression in Dementia - specific scales developed for rating severity of depression in dementia - and the Geriatric Depression Screening Scale. Methodological problems and limitations of psychopathological assessment in dementia caused by restriction of self-report, information gathered by collateral sources, manifestations of age and interference of somatic symptoms are discussed. PMID- 11894189 TI - [The round table in Karlsruhe - help for adequate housing of chronic psychic patient]. PMID- 11894188 TI - [The Tubingen questionnaire of treatment satisfaction]. AB - OBJECTIVE: Development of an instrument to assess patient satisfaction with inpatient psychiatric treatment as one component of quality of care. METHODS AND RESULTS: The TUBB 2000 has been developed in four steps (n = 80; n = 111; n = 104; n = 135) and covers the inpatients' satisfaction with 22 areas of psychiatric care. Three factors were extracted: "atmosphere", "quality of treatment", "autonomy". The relationship to the therapeutic staff was especially important for the patients. Intervention variables had no significant impact on patients' satisfaction. Answers to change oriented questions have been less variable than to a state oriented questions. CONCLUSION: The construct patients' satisfaction implies many methodological pitfalls. The TUBB 2000 can provide useful change sensitive data of patients' satisfaction with the quality of care in a psychiatric unit. It may help to increase quality of inpatient care in psychiatry. PMID- 11894190 TI - ["REGRATIO" - A vocational reintegration project for psychiatric patients]. AB - OBJECTIVE: Theoretical background and first results of a vocational rehabilitation program for unemployed persons with chronic psychiatric disabilities will be presented. The aim of the program is job-placement in the regular labour market. METHODS: After a 3-month preparatory period the participants were given the possibility of "training on-the-job" in the areas of social services, catering, and care taking activities in 12-months' rehabilitation reintegration courses in co-operation with a social service centre. In addition, a 6-months aftercare period will be included. RESULTS: More than one third of 34 participants (38 %) were placed in a regular employment. CONCLUSIONS: The results support the importance of vocational rehabilitation programs as a supplementary component of the care system of chronic mentally ill patients. PMID- 11894192 TI - [A case of kleptomania?]. AB - We report on a case of kleptomania with manifestation as deceptive ordering by catalogue. The patient is additionally suffering from severe agoraphobia. PMID- 11894191 TI - [In-patient hypnotherapeutic trauma exposure for posttraumatic stress disorder: a case report]. AB - This paper describes the treatment of a patient with the diagnoses of a borderline personality disorder and posttraumatic stress disorder (DSM-IV and ICD 10) within the setting of a psychiatric ward spezializing in depression. For purposes of controlled re-exposure to the patient's trauma, a hypnotherapeutic method was chosen. A significant reduction of symptoms, in particular the intrusions and the hyperarousal, was observed. Of great importance in the successful outcome of this case is the integration of hypnotherapy into a multi dimensional treatment concept including group therapy, physical therapy and anxiety-reducing self-management therapy. This approach facilitated the development of trust and security in the patient required for the hypnotherapeutic intervention and minimized the splitting tendencies specific to borderline patients. Further discussion is centered on the difficulties arising in the context of an emergency ward setting with its high intensity atmosphere encompassing the danger of retraumatization of this special group of patients. Finally several aspects of the clinical implications of this method are addressed. PMID- 11894193 TI - Primary Care: Is It the Setting to Address Sleep Disorders? PMID- 11894194 TI - Symptom-Based Prevalence of Sleep Disorders in an Adult Primary Care Population. AB - The prevalence of sleep disorders in a primary care physician practice in Moscow, Idaho, was studied between February 7, 1997, and February 6, 1998. This primary care clinic visit population was surveyed for this 1-year period. Every patient above the age of 18 years who visited the Moscow Clinic in this time period was either approached by our on-site researcher during the patient's clinic visit or contacted via mail. Out of a total of 1249 adult patients who met with our on site researcher during their clinic visit, 962 (77.0%) completed questionnaires and were interviewed for symptoms of sleep disorders. An additional 292 patients completed mailed questionnaires, resulting in a total of 1254 participants in the study. The percentages of patients in our sample reporting symptoms of the following sleep disorders were insomnia (32.3%), obstructive sleep apnea syndrome (23.6%), and restless legs syndrome (29.3%). This study demonstrates the need for heightened awareness and subsequent diagnosis and treatment of sleep disorders in the primary care population. PMID- 11894195 TI - Characteristics of Sleep-Related Obstructive Respiratory Disturbances in Childhood. AB - Sleep-related obstructive respiratory disturbances in childhood differ significantly from the adult's obstructive sleep apnea syndrome (OSAS). In contrast to adults, in children with OSAS the disturbance of the macrostructure of sleep, the increase of the number of apneas and hypopneas, and the diminution of oxygen saturation are not so prominent. Restlessness of the sleep, as reflected by movement arousals together with cortical (electroencephalograph recorded) arousals, is important. The combination of clinical symptoms and polysomnographic parameters is necessary to diagnose OSAS in children. PMID- 11894196 TI - Familial Aggregation and Segregation Analysis of Snoring and Symptoms of Obstructive Sleep Apnea. AB - To investigate possible modes of inheritance that would explain familial aggregation in obstructive sleep apnea (OSA), familial correlation and segregation analyses were performed on data derived from 584 pedigrees with 2019 cases enrolled in the Tucson Epidemiologic Study of Obstructive Airways Disease (TESOAD) who were at least 10 years of age and who had information pertaining to snoring and daytime sleepiness. Data were obtained from the 9th (May 1984 to October 1985) and 12th (February 1990 to October 1992) surveys of the TESOAD, which is a random, stratified sample of the non-Hispanic Caucasian population of Tucson, Arizona. A snoring phenotype was considered present if it occurred on at least some nights. A "sleep apnea" phenotype was constructed if participants snored and experienced daytime sleepiness. Familial correlations for snoring showed significant mother-child and sibling correlations but not father-child correlations. For sleep apnea, significant parent-daughter but not parent-son or sibling correlations were observed. Segregation analyses for snoring with regressive familial effects and sibling, age, and obesity covariates showed no evidence for mendelian transmission. However, additional familial effects were present that suggested phenotype aggregation from polygenic or environmental factors, or both. For the sleep apnea phenotype, similar segregation analyses indicated that mendelian dominant or codominant models were possible. However, the analyses also suggested that a nongenetic model fit the data as well. In addition, consistent with the familial correlations, specific maternal- and sibling-related effects remained even after inclusion of age, gender, and obesity covariates. These data support the concept that inheritable or shared environmental factors contribute to the development of OSA and that maternal components may be more important than paternal ones. PMID- 11894197 TI - Human Sleep Apneas and Animal Diving Reflexes: The Comparative Link. AB - Adaptations to survive periods of limited access to oxygen should have been favored along the evolution of vertebrates. Paradigmatic examples of this adaptation are the diving animals, which can sustain prolonged and repetitive periods of anoxia. These animals support what would be considered a severe gas imbalance in their internal environment thanks to three main strategies: increased oxygen stores, resistance to asphyxia, and reduced metabolic expenditure during the apneic intervals. However, diving animals developed their abilities from very old life-sustaining responses that should have been used on many other occasions. Humans with sleep apneas perhaps share many physiological adaptations with diving animals. We review here the extent of such similarities and offer clear evidence of its existence and suggest possible research lines that could improve the clinical knowledge about this condition. PMID- 11894199 TI - SITE-SPECIFIC VERSUS DIFFUSE TREATMENT/PRESENTING SEVERITY OF OBSTRUCTIVE SLEEP APNEA. PMID- 11894198 TI - Another Wall to Tear Down: Instructions from Berlin. PMID- 11894200 TI - Maxillomandibular Advancement (MMA) in a Site-Specific Treatment Approach for Obstructive Sleep Apnea: A Surgical Algorithm. AB - Maxillomandibular advancement (MMA) is the most successful acceptable surgical treatment, excluding tracheostomy, for obstructive sleep apnea syndrome (OSAS). Nevertheless, the indications for and staging of MMA, with respect to the many procedures available, are unsettled and often limited to severe OSAS, dentocraniofacial deformities, and when other surgeries have failed. An algorithm is presented that defines the indications for MMA in an expanded role, based on a site-specific approach, according to proposed principles that include general goals and guidelines for governing the surgical treatment of OSAS. PMID- 11894201 TI - Tongue Base Reduction with Radiofrequency Tissue Ablation: Preliminary Results after Two Treatment Sessions. AB - Over the last few years, different surgical techniques for the treatment of obstructive sleep apnea syndrome have been developed. While new methods for the treatment of velopharyngeal obstruction turned out to be safe and effective, treatment of hypopharyngeal obstruction due to tongue base hypertrophy has remained, in many aspects, an unsolved problem. Surgical techniques for partial resection of the tongue base (midline glossectomy, lingualplasty) are effective but very invasive procedures requiring temporary tracheotomy, and have high postoperative morbidity. A maxillofacial approach showed significant reduction in the Respiratory Disturbance Index (RDI), especially when bimaxillar osteotomies are performed. Along with the concerns of postoperative morbidity, these techniques require general anesthesia and hospitalization. Tongue base reduction with temperature-controlled radiofrequency tissue ablation was introduced in 1998, and has proven to be a safe and simple procedure. Significant reduction in RDI has been shown in the majority of the treated patients. This procedure does not require general anesthesia and has low postoperative morbidity. For curative results, tongue base reduction with radiofrequency requires multiple treatment sessions. The goal of the present study is to investigate the beneficial effect of increased amount of energy applied per treatment session, reducing the number of treatment sessions per patient. By delivering increased amounts of energy, a similar cure and/or responder rate with 2 treatment sessions, rather than 5 to 6 as published in earlier studies, was accomplished. Postoperative morbidity was similar to previously published results although there was a slight increase in postoperative complications. Apart from the reduced number of treatment sessions needed per patient, we also demonstrated the beneficial effect of prophylactic use of antibiotics in our study group. This more aggressive treatment scheme appears to be well tolerated by the patients and may be an effective means of reducing the overall number of treatments. PMID- 11894203 TI - Improvement of CPAP Therapy by a Self-Adjusting System. AB - The first generation of Auto CPAP devices caused respiratory arousal by apnoes, hypopnoeas, incomplete obstructions and pressurechanges. The new, second generation of CPAP devices which is based on forced oscillation technique will change the pressure with slower velocity and before the respiratory arousal reaction will occur (1, 9, 10). Fifty patients with severe sleep apnoea (AHI 66+/ 26 /h) were treated with both, constant- CPAP (continous positive airway pressure) or Auto CPAP under polysomnographic control in a randomised order. The Auto CPAP based on forced oscillation technique reduced the number of apnoeas and hypopnoeas as did most of the other Auto CPAP systems to AHI 2.5+/-5.9 /h (p<0.05). In comparison to Auto CPAP of the first generation it also decreased the number of respiratory arousal reactions caused by apnoeas and hypopnoeas. However there is still a significant difference to number of arousal detected with constant CPAP (p<0.01). In conclusion although the new generation of Auto CPAP reduced the number of respiratory arousals compared to first generation, we did not find a therapeutical benefit for patients with severe SAS. PMID- 11894202 TI - Effect of Jaw and Head Position on Airway Resistance in Obstructive Sleep Apnea. AB - This study evaluated whether changes in jaw and neck position caused substantial airway resistance (Raw) changes in normal controls and obstructive sleep apnea (OSA) subjects. Subject groups included 12 male nocturnal polysomnographically diagnosed OSA patients and 16 healthy male control subjects. Raw was assessed plethysmographically and measured under the following conditions: neutral head posture with 0/3, 1/3, 2/3, or 3/3 of the subjects maximum forward jaw position; normal jaw (0/3 forward) with fully flexed, extended, right or left rotated head position. Both groups showed a similar significant decrease in Raw upon jaw protrusion. OSA patients showed a significantly higher baseline (normal jaw, neutral head posture). Raw and both subject groups also had a clear increase in their airway resistance with flexion and to a lesser extent with neck rotation and extension. These data document that airway resistance can be significantly influenced by head and jaw positioning with protrusion of the jaw reducing Raw and flexing the neck increasing Raw. PMID- 11894204 TI - Physical Exercise as an Adjunct Therapy in Sleep Apnea-An Open Trial. AB - BACKGROUND: The aim of this study was to determine in an open trial if physical exercise in sleep apnea patients is safe and/or influences respiratory disturbance index (RDI). METHODS: After being treated 3 months or more with nasal CPAP for moderate to severe sleep apnea syndrome, eleven patients (1 female, 10 male, mean age 52.2 years) began a six-month period of supervised physical exercise twice a week, 2 hours each time. Before and after this period a Polysomnography without CPAP was recorded, along with a bicycle exercise test with lactate profile, echocardiography, body-weight, and body-height measurement. RESULTS: No adverse effects or cardiopulmonary problems were observed. There was no significant change in body weight with physical training; no significant difference in either min SaO2 nor mean SaO2; and no significant improvement in fitness. No adverse cardiopulmonary effects or problems were observed. There was a decrease of the RDI from 32.8 to 23.6 (p < 0.05), without a significant change in the REM-sleep portion of total sleep time (TST), NREM sleep, or TST. CONCLUSIONS: A prescription for mild to moderate exercise is safe in the management of sleep apnea, and, even in the absence of a fitness improvement, there occurred a decrease in RDI without a change in sleep architecture. PMID- 11894205 TI - Diagnostic Approaches to Childhood Obstructive Sleep Apnea Hypopnea Syndrome. AB - Obstructive sleep apnea hypopnea syndrome (OSAHS) is a common sleep disorder in adults that is increasingly recognized in children, affecting 1 to 3% of children. Children experience a spectrum of severity related to the degree of upper airway obstruction, the duration of the disease, and the presence or absence of hypoxemic episodes. Failure to diagnose and treat OSAHS can result in serious, but generally reversible consequences for the child including impaired growth, neurocognitive and behavioral dysfunction, and cardiorespiratory failure. Even mild OSAHS appears linked to reversible health consequences. Adenotonsillar hypertrophy is the major predisposing factor for OSAHS in childhood. However, enlarged tonsils and adenoids can be a normal finding in young children and are not diagnostic for OSAHS. The identification of children with OSAHS is often difficult because affected children may have no signs or symptoms when awake. Furthermore, clinical assessment cannot reliably distinguish between simple snoring and OSAHS. Adenotonsillectomy is the most common therapy for OSAHS in children, but surprisingly, only a small percentage of children undergo any diagnostic testing prior to surgery. Thus, the challenge is to develop new diagnostic strategies that effectively screen, identify, and treat children most likely to benefit from specific treatment. PMID- 11894206 TI - [MR-angiography and duplex-ultrasonography: predictive reliability for angiographically determined internal carotid artery stenosis >/= 70-99%]. AB - PURPOSE: A prospective study was undertaken to evaluate whether digital subtraction angiography (DSA) which is still associated with a substantial morbidity can be replaced by less invasive diagnostic modalities such as duplex scanning (DS) and magnetic resonance angiography (MR-A) for the detection of angiographically defined internal carotid artery (ICA) stenosis >/= 70 %. PATIENTS AND METHODS: A total of 47 patients with suspected severe ICA stenosis underwent examination of their carotid arteries using duplex studies, MR-A and DSA. According to the study protocol, the arteriographic diameter reduction (DR) >/= 70 % which had to be predicted by DS and MR-A was determined following the NASCET criteria. RESULTS: Stroke rate following DSA amounted to 2.1 %. In 94 carotid arteries studied by DSA 34 times a DR >/= 70 % was found. Using ROC curve for determining optimal discriminant value, duplex-derived peak systolic velocity (PSV) >/= 250 cm/s provided a sensitivity of 94.1 %, a specificity of 80 %, a positive predictive value (PPV) of 72.7 % and a negative predictive value (NPV) of 96 % to characterise an ICA stenosis >/= 70 %. Due to an inadequate PPV, PSV failed to suffice as the sole preoperative diagnostic modality even if different PSV velocity cut points were applied. On the other hand, end diastolic velocity (EDV) >/= 150 cm/s provided a PPV of 100 % thereby identifying 16/34 ICA stenoses >/= 70 % in our study. MR-A showed a sensitivity of 91.2 %, a specificity of 88.3 %, a PPV of 81.6 %, and a NPV of 94.6 % to predict an ICA stenosis >/= 70 %. CONCLUSION: In our series, both duplex-derived PSV as well as MR-A provided high sensitivity to detect surgically relevant ICA stenosis. However, to select patients for surgery inclusion of EDV proved to be important due to a high PPV and may spare conventional angiography half of patients with stenosis exceeding 70 %. PMID- 11894208 TI - [Injury of polyester grafts by vascular clamps]. AB - Protected vascular clamps are not new. Clamp associated damage of human arteries has already been published over 20 years ago. The necessity of protective clamps seems to have been forgotten. In our explant archive (230 explants) we have observed an accumulation of graft ruptures in the groin (13 of 25 ruptures). We presume a multifactorial process. Clamp damage could be part of it. The aim of this study is to prove the clamp induced damage of polyester vascular grafts and to examine whether protected clamps can reduce this. METHOD: Five unprotected (Aesculap(R) FB512R, FB502, FB517, Ulrich CC1235, CV3535) and 5 protected vascular clamp types (Aesculap(R) FB667, FB668, Edwards(R) - formally Baxter(R) - Fogarty(R) CV5050, CV5201, Edwards(R) Cosgrove(R) CV1033) were tested. A longitudinal burst test was performed after maximal clamp closure on 6 different, multifilament polyester yarns of 2 different vascular grafts manufacturers (B. Braun(R), Edwards(R)). RESULTS: The yarn tests with protected clamps showed no difference to those of the unclamped yarns. After clamping with unprotected vascular clamps the stress-strain-diagrams differed significantly. The mean, maximum burst strength was up to 75 % lower. Video documentation revealed filament ruptures. Damage of the yarn surface was seen on a simple woven graft in scanning electron microscopy (SEM). DISCUSSION: The application of unprotected vascular clamps on polyester vascular grafts is common in Germany (56 %). The observed damage of multifilament polyester yarns makes it necessary to re consider the use of unprotected vascular clamps. The benefit for biological vessels has already been shown. PMID- 11894209 TI - [Late complications in the aorta and iliac artery following open aortic surgery]. AB - Late complications following conventional repair of abdominal aortic surgery were measured in 304 patients. Further operations were necessary in 4.6 % of the patients after an average time of 8 years. Indications for further operation were: Suture line aneurysms (6), endoleak (2), kinking of the prosthesis (2), occlusion (2), infection (2). In one of these cases an aortoduodenal fistula was observed. There was no lethality in the redo operations. It can be shown that vascular follow up operations are necessary after conventional aortic surgery. In comparison to endovascular surgery, however, they are less frequent and can be performed with acceptable security for the patient. PMID- 11894210 TI - [Endovascular therapy in combination with conventional vascular surgery for the treatment of peripheral arterial obliterative disease]. AB - INTRODUCTION: In a prospective trial we analysed the results and the management of complications following a combined endovascular and vascular surgical therapy of peripheral occlusive arterial disease (POAD). PATIENTS AND METHODS: From November 1998 until January 2001 a total number of 61 patients with 64 extremities were included in this study. By preoperative angiography 19 patients had stenoses at three levels, 42 patients suffered from stenoses at two levels. The iliac axis was recanalized by intraluminal angioplasty (ITA) plus stent placement under general anesthesia. Simultaneously an infrainguinal bypass reconstruction and a local thrombendarterectomy (TEA) rsp. were performed. Intra- and postoperative complications and the patency rates as assessed by colour doppler ultrasound and angiography were analysed. RESULTS: The rate of conversion from endovascular to conventional surgery was 12.5 %. In 56 cases the endovascular therapy of the iliac axis was successful. In 28 patients a distal bypass was implanted, in 25 patients a local TEA was performed. Intraoperatively 6 dissections (10.7 %) were noted, dislocation of stents were seen in 4 patients (7.1 %), perforations occurred in 2 patients (3.6 %). Both perforations and 5/6 dissections were detected intraoperatively and were treated by endovascular means without complications. The early postoperative patency rate was 98.2 %, the secondary patency rate was 100 % and the cumulative patency rate after two years was 98.2 %. DISCUSSION: ITA and stent placement in the iliac axis can be established quickly and safely by the vascular surgeon. Intraoperative complications can be managed by endovascular means in most cases. Stent dislocation is avoidable in most cases. The complication rate after such combined endovascular therapy and conventional vascular surgery is determined by the surgical but not the endovascular part. Excellent early results and low complication rates lead to the conclusion that endovascular therapy in combination with conventional vascular surgery seems to be a reasonable supplement to the therapeutic options for the treatment of POAD. PMID- 11894211 TI - [Combination of endovascular procedures with open vascular surgery]. AB - The combination of open surgery with endoluminal catheter techniques offers new chances in the treatment of multi-staged arterial occlusive disease. We report our experiences in 205 patients. In n = 125 patients TEA or bypass were undertaken in the groin with simultaneous catheter intervention proximally or distally. In 31 patients the operation was carried out above knee and in 49 cases below knee. 94 patients with an open operation in the groin and simultaneous iliac catheter interventions are in the follow up. For these interventions the combined secondary patency rate was 77 % after three years. PMID- 11894212 TI - [Primary and secondary aortoiliac reconstructions in patients with coexistent horseshoe and pelvic kidney]. AB - The coexistence of dystopic kidney and abdominal aortic aneurysm or aortoiliac occlusive arterial disease is uncommon (0.12-0.15 %) and constitutes a technical challenge to vascular surgeons. Between 1997 and 1999 we performed 249 aortoiliac reconstructions and encountered four patients (1.6 %) with dystopic kidneys. The aortoiliac reconstructions were performed successfully in all patients while maintaining renal blood supply. Aortoiliac reconstructions in patients with horseshoe or pelvic kidney show similar results as regular reconstructions on the condition that optimal preoperative diagnosis and operative technique are used considering atypical renal vessels. Nevertheless, pelvic kidneys can lead to serious complications as we describe in this report. On the basis of our patients anatomy, embryology and surgical management of this entity. PMID- 11894213 TI - [False aneurysm of the popliteal artery after implantation of an alloplastic bypass: surgical treatment]. AB - The false aneurysm of the popliteal artery is a rare complication in bypass surgery after the implantation of an alloplastic prosthesis. We saw this complication in 0.7 % of our patients. We report on 6 cases successfully treated by surgery. The false aneurysm developed within 2 weeks up to 44 months following the operation. We describe the symptoms, the details of diagnosis and of surgical treatment. In most cases the false aneurysm is symptomatic and should be operated immediately. PMID- 11894214 TI - [Ambulatory vascular exercise training in Dortmund]. AB - Peripheral arterial occlusive disease is chronic and progressive. One of the reasons is lack of movement. The pain-free walking distance can be increased permanently through walking exercises described in the guidelines of the German Society for Vascular Training. Form and order of the training are described. The pleasure in movement and preservation of the own activity increases the motivation of the participants. PMID- 11894215 TI - [Patency of surgically revised ePTFE-dialysis access grafts]. AB - Native av-fistulas are the access of first choice for long-term hemodialysis. However, a large number of patients require an alternative vascular access, e. g. ePTFE grafts. Patency of ePTFE grafts is inferior to that of native av-fistulas. PURPOSE: To analyse the effectiveness of surgical revisions of occluded straight ePTFE dialysis access grafts. METHODS: Retrospective review of all upper arm dialysis access procedures from 1/94 to 8/99. RESULTS: Redo surgery was performed in 67 patients. Av-fistula dysfunction was caused by venous anastomotic stenoses (22 %), outflow occlusion (9 %), arterial anastomotic stenoses/inflow occlusion (12 %), and intragraft stenoses (6 %). 9 grafts had to be revised due to infection or perigraft hematoma (14 %). In 37 % the cause of graft occlusion could not be identified. Neither the cause of occlusion nor the type of treatment correlated with patency after revision. 6- and 12-months primary patency after surgery were 29 % and 11 %. 59 shunts required up to 12 revisions to maintain patency. Thus, secondary 1 yr-patency after revision was 29 %. CONCLUSION: Patency after redo surgery is disappointing. However, with repeated procedures ePTFE grafts remain open > 1 year in 29 % of the patients. PMID- 11894216 TI - [Acceptance of computer-assisted surgery planning in visceral (abdominal) surgery]. AB - The acceptance of computer assisted surgical planning was studied by polling 500 German surgeons. 500 questionnaires were sent, from which 105 have been returned, from which 102 could be analysed. The questionnaire consisted of 14 questions relating to the current practice of surgery planning as well as to desired improvements. In general, the computer assisted surgical planning was assessed as a promising tool in the future. 82 % of the surgeons expected a benefit by computer assisted surgical planning. Better assessment of resectability, better training of residents and more safety of surgical procedures were mentioned in the majority. Computer assisted surgical planning should focus on 3D visualization of spatial relations between lesions and vascular structures, on facilities for the preoperative trial of resection strategies and on the documentation of the planned procedure. The current version of a research prototype (Surgery-Planner) is presented which is dealing with these goals. PMID- 11894217 TI - [Mobile autonomous robots-Possibilities and limits]. AB - Besides industrial robots, which today are firmly established in production processes, service robots are becoming more and more important. They shall provide services for humans in different areas of their professional and everyday environment including medicine. Most of these service robots are mobile which requires an intelligent autonomous behaviour. After characterising the different kinds of robots the relevant paradigms of intelligent autonomous behaviour for mobile robots are critically discussed in this paper and illustrated by three concrete examples of robots realized in Lubeck. In addition a short survey of actual kinds of surgical robots as well as an outlook to future developments is given. PMID- 11894218 TI - [Management of aneurysms of peripancreatic arteries--2 case reports]. PMID- 11894219 TI - [Arterial port-catheter dislocation into the duodenum]. AB - Arterial port-catheter dislocations into the duodenum are rare complications. We report on a 64-year-old male with sigmoid-resection for carcinoma, left hemihepatectomy and arterial port-catheter implantation into the common hepatic artery for liver metastases in both lobes and partial dislocation of the port system into the duodenum. Therefore, the port-catheter system had to be explanted after 8 cycles of cytostatic therapy and the duodenum had to be closed by a suture. Six months after surgery the patient felt well, a new singular metastasis in the right liver lobe could be treated successfully by laserinduced thermotherapy. At present the patient has no evidence for another metastasis. PMID- 11894220 TI - [Unusual complications of the Peutz-Jeghers-syndrome in two consecutive generations of the same family]. AB - The Peutz-Jeghers syndrome is an autosomal dominant inherited disease, characterized by the presence of hamartomatous polyposis of the gastrointestinal tract and perioral mucocutaneous pigmentation. The incidence of surgical complications in these patients is relatively rare, and correlates with the size and location of the polyps. We report on two complications of the Peutz-Jeghers syndrome which occurred in two generations of the same family. There was a perforation and an invagination of the small intestine. Both cases were treated by resection of the small intestine. PMID- 11894223 TI - Trends in vision and hearing among older Americans. PMID- 11894222 TI - FOXP2 is not a major susceptibility gene for autism or specific language impairment. AB - The FOXP2 gene, located on human 7q31 (at the SPCH1 locus), encodes a transcription factor containing a polyglutamine tract and a forkhead domain. FOXP2 is mutated in a severe monogenic form of speech and language impairment, segregating within a single large pedigree, and is also disrupted by a translocation in an isolated case. Several studies of autistic disorder have demonstrated linkage to a similar region of 7q (the AUTS1 locus), leading to the proposal that a single genetic factor on 7q31 contributes to both autism and language disorders. In the present study, we directly evaluate the impact of the FOXP2 gene with regard to both complex language impairments and autism, through use of association and mutation screening analyses. We conclude that coding region variants in FOXP2 do not underlie the AUTS1 linkage and that the gene is unlikely to play a role in autism or more common forms of language impairment. PMID- 11894224 TI - Trends in causes of death among the elderly. PMID- 11894225 TI - The oral health of older Americans. PMID- 11894226 TI - The changing profile of nursing home residents: 1985-1997. PMID- 11894227 TI - Robert Burns: toothache and other pains. PMID- 11894230 TI - Treasures of the College. PMID- 11894231 TI - Nurses in front line against influenza in indigenous communities. PMID- 11894232 TI - Tourniquets pose infection risk. PMID- 11894233 TI - UK flu crisis worsens. PMID- 11894234 TI - Private health insurance: should you join? PMID- 11894235 TI - Child's recovery gives birth to the Cancer Natural Therapy Foundation. PMID- 11894236 TI - Addressing Medicare spending. PMID- 11894237 TI - Prevention is better than cure. PMID- 11894238 TI - Learning Buteyko's breathing method changed my life. PMID- 11894240 TI - Make quality a reality in aged care. PMID- 11894239 TI - Gaughan v Bedfordshire Health Authority. PMID- 11894241 TI - Latex allergy is not to be sneezed at. PMID- 11894242 TI - Burn wounds: assessment and first aid treatment. PMID- 11894244 TI - Perioperative nursing in the third millennium. PMID- 11894245 TI - What's new in the world of the perioperative nurse surgeon's assistant. PMID- 11894251 TI - What's in a name? PMID- 11894253 TI - Skin diseases in older people. PMID- 11894254 TI - Rural health research: have we turned the corner? PMID- 11894255 TI - Researching the rural-metropolitan health differential using the 'social determinants of health'. AB - Recent research indicates that the health status of rural people is inferior to that of people living in metropolitan Australia. This paper summarises the rural metropolitan health differential and turns to the field of research being called the social determinants of health for explanations of rural health inequalities. The paper explores the ways in which psychosocial factors can interact with material, behavioural and sociocultural factors to contribute to health outcomes. It suggests that the concepts of place and rurality may be useful in future research on the determinants of population health. Further research issues are identified that need to be addressed if we are to understand the complexities of rural health disadvantage. PMID- 11894256 TI - Rural health research in Canada: at the crossroads. AB - Rural health research in Canada is at the crossroads. Jolted by the establishment of the Canadian Institutes of Health Research, rural health researchers are trying hard to overcome past benign neglect and the lack of cohesion and collaboration within the rural health research community. Although there is considerable catching-up to do, rural health research in Canada has a firm foundation. Backed by a growing network of rural health research centres, researchers are searching for ways to work together in order to advance rural health research and the health and wellbeing of rural Canadians. PMID- 11894257 TI - Nursing research in practice: the case study revisited. AB - This paper makes reference to some of the many issues and problems surrounding nursing research in the context of the rural and remote areas of Australia. The focus is on the use of case studies as an appropriate methodology for small-scale research projects and for students preparing for examinations at higher degree levels. Throughout this paper, the term rural will be used to denote rural and remote area nursing. This is for convenience and does not deny the special qualities of rural or remote area nursing. PMID- 11894258 TI - An assessment of funding to support rural and remote health research in Australia. AB - A systematic search was undertaken to ascertain the nature, source and extent of funding awarded to research projects that were directed specifically at aspects of rural health over the past decade. Comment is also made on the challenge of obtaining such information directly from databases. The sources investigated were the conventional research funding bodies, hospital trusts and foundations, university funding schemes and government sources. The results of these searches revealed a crude average of 3 million dollars per year from conventional research funding with the remaining sources adding a similar amount in total. Analysis of the data using a framework modified from the Strategic Review of Health and Medical Research in Australia shows that funding is concentrated in the health services and public health areas with a preponderance of funding being directed towards the description of conditions and interventions. Significant levels of funding have been directed towards the National Health Priority Areas. PMID- 11894259 TI - The emergence of rural health research in Australia. AB - The late 1980s saw a renewed interest in rural health in its own right with State jurisdictions establishing defined rural health units. There was recognition that health professionals in rural communities often needed training to fulfil their roles in rural and remote communities. Simultaneously, the number of regional universities increased. A number of such universities grouped to form a consortium and were funded through the Australian Health Ministers' Advisory Council (AHMAC) to form the Australian Rural Health Research Institute. Consequently, a series of conferences specifically addressing rural health research needs occurred, and the Australian Journal of Rural Health was established. Through both of these avenues a trend towards more research presentations and publications can be seen. The National Health and Medical Research Council, under the auspices of AHMAC, has provided an overview of the state of rural health research in Australia and developed recommendations, which form the framework for a future strategy for rural health research. These recommendations have been endorsed by AHMAC, which has now funded the initiation of the implementation of the strategy. PMID- 11894260 TI - Systematic overview of rural health research in Australia published in the serial literature. AB - It is acknowledged that research outcomes need to be made available to other researchers, practitioners and consumers if the benefits of such research are to be disseminated as quickly as possible to other researchers and translated to influence practice and produce new products. If strategies are to be developed on future research directions, an analysis of current output will provide baseline data as well as a map for analysing gaps and overlaps in the research effort. This paper provides an overview of rural health research output in serial publications from 1990. It highlights the content of such literature, the relationship to the National Health Priority Areas and the types of research produced. The overview showed that there was a yearly increase in the absolute number of defined research articles produced over the period. There was also an increase in the relative number of rural health articles that could be defined as research articles (as opposed to commentary-type articles). Forty per cent of research articles are in the National Health Priority Areas and more than 30% address indigenous health specifically. Fifty per cent of articles address public health or health services issues. These data provide a general picture of the type and content of Australian rural health research that has been reported in the serial literature over the decade. This can be used to inform future strategies, provide the baseline for future targets, as well as giving a general picture of the gaps and overlaps from which more specific data can be compiled. PMID- 11894261 TI - Nurses can help protect Medicare. PMID- 11894262 TI - Caring for kids on remand. PMID- 11894263 TI - Crisis. What crisis? The myth of Medicare chaos. PMID- 11894264 TI - Seeking lessons in medication errors. PMID- 11894265 TI - Aromatherapy for health professionals. PMID- 11894266 TI - St Luke's specialises in palliative care education. PMID- 11894267 TI - Side position best. PMID- 11894268 TI - Back to back again. PMID- 11894270 TI - Nappy prevents premmie posture problems. PMID- 11894269 TI - The physiology of pain. AB - To manage pain effectively, clinicians need to have an understanding of the processes involved in the facilitation of pain transmission from the site of injury to the CNS and processes that augment or inhibit that transmission. Significant advances in our under-standing of the complex and integrated communication system involved in pain transmission have supported the use of a veritable armory of pharmacological and non-pharmacological methods to reduce the individuals' perception of pain. Comprehensive assessment and reassessment of pain remains the single most important determinant in good pain management. Through an integration of the fundamental knowledge regarding the physiology of pain transmission and the skills required for assessment of the multidimensional nature of an individual's pain, the nurse plays a central role in ensuring timely and effective strategies are employed for optimal pain management. PMID- 11894271 TI - Nursing in a foreign country. PMID- 11894272 TI - Addressing aboriginal health inequalities. PMID- 11894273 TI - Creating a culture of zero tolerance. PMID- 11894274 TI - Silence on violence no longer. PMID- 11894275 TI - Nurses teaching nurses. PMID- 11894276 TI - Haemochromatosis: work and social implications. AB - Hereditary haemochromatosis is prevalent in our society and requires vigorous and lifelong treatment. Diagnosis of the disorder may be delayed or missed due to its nonspecific symptoms and diverse manifestations. This can have lifelong consequences for the sufferer. Early diagnosis prevents severe organ damage and premature death and leads to a normal productive life. Screening family members will enable early diagnosis of those with the HFE gene mutation. The stigma attached to having a genetic condition may have consequences for job opportunities and for life and health insurance. Education of the community (including workplaces and employers) as well as health service providers will not only increase the likelihood of early diagnosis of haemochromatosis but will ease the social consequences of suffering from this disorder. PMID- 11894277 TI - Perspectives on living with ovarian cancer: young women's views. AB - Ovarian cancer is the fourth leading cause of cancer-related deaths in women. Ovarian cancer, and its treatment, has a considerable effect on the quality of life of women diagnosed with the disease. Young women diagnosed with ovarian cancer must confront life-threatening illness at a time when many are in the midst of raising children, maintaining a household, and actively engaging in work and career activities. Very little has been reported about the perspectives of young women regarding their experiences with ovarian cancer. This article reports data from 39 women 45 years of age or less concerning the impact of ovarian cancer and its treatment as well as the availability of support. At the time of the study, the women were, on average, 38 years of age and approximately two thirds were married and had children. About half of the women were working. The most frequently identified problems included side effects (n = 25), fear of recurrence (n = 25), and difficulty sleeping (n = 25). On average, women reported experiencing 10.4 problems since diagnosis. Of those who experienced problems, less than 50% perceived they had received adequate help. Approximately two-thirds of these women experienced a lifestyle change. Quality of life was rated significantly lower following their experience with ovarian cancer. Implications for oncology nurses emerge in areas of assessment, referral, and patient teaching. PMID- 11894278 TI - A community intervention plan to prevent skin cancer in male golfers. PMID- 11894280 TI - The human genome project--what will we do with it? PMID- 11894279 TI - [Reinforcing the educational mandate on pain of the ACIO (Canadian Association of Oncology Nurses)]. PMID- 11894281 TI - Dealing with missing data: a common problem in nursing research. PMID- 11894283 TI - Immigrants and equitable health-care delivery in rural areas. AB - In response to settlement patterns in Australia, most immigrant specialist services and programs have been developed in metropolitan locations and large provincial cities. However, immigrants have also settled in smaller numbers in country locations. It is of concern, therefore, to consider how responsive and equitable health-care services can be delivered in country regions when immigrants do not reach the critical mass that would warrant the development of specialist services. This paper draws on a consultation conducted in South Australia to propose a way forward in linking country health services with local immigrant communities and immigrant specialist services in cities. PMID- 11894282 TI - Testing control of radiation-induced diarrhea with a psyllium bulking agent: a pilot study. AB - Sixty cancer patients who were undergoing radiation therapy to the pelvis of at least 4,000 cGy in 20 fractions over four weeks were randomized to take or not take Metamucil. Results were analyzed for the presence of radiation-induced diarrhea in two groups: patients taking Metamucil (n = 30) or not taking Metamucil (n = 30). The Murphy Diarrhea Scale was developed to assist in the synthesis of data collected in daily patient-reported diaries. Results were analyzed using ANOVA F-tests. Metamucil significantly decreased the incidence (p = 0.049) and severity (p = 0.030) of diarrhea and showed a strong trend in reducing the use of anti-diarrhea medication (p = 0.062). According to this pilot study, Metamucil was an effective method of controlling radiation-induced diarrhea. Results of this pilot study have implications for clinical practice and nursing research. PMID- 11894284 TI - Patterns of use of complementary health services in the south-west of Western Australia. AB - The objective of this research was to identify patterns of complementary health service usage by rural Western Australians in the south-west of the State. Complementary health providers identified by health users included homeopaths, chiropractors, naturopaths, acupuncturists, faith healers, herbalists, reiki or energy workers, counsellors, physiotherapists, osteopaths, podiatrists and reflexologists. More than half of the health provider usage in the region was with complementary therapists and the remainder with medical doctors. The main reason identified for using complementary therapists was their level of skills and the main reason identified for not using them was a lack of knowledge about what their services could provide. PMID- 11894285 TI - Structures of prevention: a safe-sex/safe-injecting audit of Mount Alexander Shire, a methodological pilot. AB - This paper reports the findings of a methodological pilot project that examined the availability of safe-sex and safe-injecting products in a rural shire in central Victoria. The study mapped the distribution and audited the availability of condoms, water-based lubricants, needles/syringes and needle disposal units to provide baseline data on access to and the availability of products that are directly related to the prevention of the human immunodeficiency virus, sexually transmissible infections and blood-borne viruses. Clear disparities were observed in the availability and pricing of safe sex/safe use materials between regional centres and outlying areas in the Shire. The findings make it clear that an emphasis on overcoming structural impediments to the adoption of safe sex/safe injecting practices is required. PMID- 11894286 TI - Health beliefs and perceptions of women presenting or not presenting for mammographic screening in a rural health setting. AB - Breast cancer continues to be a major health concern among Australian women. Recently, free mammography screening has been offered as a joint State and Commonwealth initiative to enable early detection. This program has particular significance in rural areas where access to health-care facilities, particularly those of a specialist nature, is limited. Attendance for screening is critical to the success of this type of program. Several lines of evidence suggest that health beliefs play a major role in compliance with recommended health behaviour. The present study investigated the role of five health beliefs: response efficacy, seriousness, concern, susceptibility and barriers to the likelihood of attending the North Coast Breast Screening Program in northern NSW. A questionnaire that measured these health beliefs was completed by 127 women who attended breast screening and 185 women who knew about the service but had not attended. Compared with those who had not undergone mammography, those presenting for screening were more health conscious, more likely to have had a mammography previously, more aware that mammography reduced the risk of developing severe breast cancer and less concerned about having a mammogram. A number of barriers to having a mammogram were also identified. The present study raises a number of concerns regarding the level of knowledge about breast self-examination and mammography as important preventive health measures among women in a rural setting. PMID- 11894287 TI - A nurse practitioner-led farmers' health service: setting up and evaluating a UK project. AB - The farming community in the UK have significant unmet health-care needs that have traditionally remained invisible. The project that is the subject of this paper is an attempt to improve access to health care for farmers and make their needs visible. It is based in an upland area of England and involves two nurse practitioners (NPs) and two support workers who provide health care for the farming community. The main targets of the project are farm accidents, mental health and occupational diseases. The NPs visit farms, on request, for consultations and also attend auction marts, agricultural shows and other farmers' meetings. Evaluation is by a telephone follow-up interview with a sample of clients seen by the NP, who will be compared with a group from a similar area who do not receive the service. A farm accident survey is underway, case notes are being audited and the NPs are keeping a reflective diary as further sources of data. PMID- 11894288 TI - Mechanics of an educational exchange. AB - Rural general practitioners occasionally exchange practices internationally. The process of exchanging is a complicated one, which involves multiple bureaucarcies. Exchanging is not for the anxious or impatient. We exchanged with a family physician in Canada and had to arrange registration, provider numbers and immigration for our Canadian colleague, as well as for ourselves in Canada. In addition, there are many personal details (cars, house, etc.) that must also be sorted out. PMID- 11894289 TI - Is Australian rural practice changing? Findings from the National Rural General Practice Study. AB - The National Rural General Practice Study (NRGPS) was the first comprehensive national study covering rural and remote general practitioners throughout Australia. It was undertaken in 1996-1997 and drew on data from existing sources such as the Australian Bureau of Statistics and the Australian Institute of Health and Welfare, together with a postal survey of general practitioners in rural and remote areas. There was a 75% response rate to the survey, which covered professional issues, personal and social issues, personal background, patient issues, recruitment and retention programs and changing health services. Overall, the study findings confirmed those of previous individual State-based studies in the early 1990s and showed that there had been some changes since those previous studies. In particular, access to continuing medical education has improved, the rural medical workforce appears to be ageing, the proportion of women rural doctors is increasing and the projected length of stay in rural practice is decreasing. Whereas in the early 1990s the projection for rural doctor numbers was continuing decline, the NRGPS projected overall numbers in rural practice as staying approximately the same over the next 5 years. In the light of these trends, the challenge is to implement targeted initiatives that improve the recruitment and retention of rural and remote general practitioners. PMID- 11894290 TI - Interpractitioner communication: telephone consultations between rural general practitioners and specialists. AB - The results of a study that sought to investigate the utility of and satisfaction with telephone consultations from the perspective of general practitioners and specialists are reported. Semi-structured interviews with rural general practitioners and specialists were used to elicit information about their most recent telephone consultations. The telephone was found to be an important means of communication for rural practitioners, primarily in terms of organising referrals. General practitioners tended to called specialists who they knew and appeared to have fairly well-formed networks of specialists who they called for most of their concerns. Trust is an important element of interpractitioner communication as it increases understanding and confidence in the reliability of the information exchanged. Good working relationships ensure that rural general practitioners have an accessible source of acceptable specialist support. PMID- 11894291 TI - The patient transit assistance scheme: a consumer's perspective. AB - This article presents findings from State-wide research in Queensland on experiences with the Patient Transit Assistance Scheme (PTAS) from the point of view of patients with a haematological malignancy who travelled for specialist treatment. The findings indicate that the PTAS is of crucial importance to patients and their families and goes some way to buffering the financial hardship incurred from relocation. However, there are also strong indications that there is insufficient community awareness of the scheme, with the consequence that it is not appropriately accessed by those in need. The results confirm previous research that points to the hardship, both financial and emotional, associated with the experience of relocation for specialist treatment. PMID- 11894293 TI - Looking forward to the challenges. PMID- 11894292 TI - Non-government organisations to combine for work on chronic disease in Aboriginal peoples, Torres Strait Islanders and rural and remote populations. AB - As a result of the National Health Priority Area Report on Cardiovascular Health and in particular, its remote and indigenous section, a consortium of five organisations ran a national workshop in Townsville in October 1999 on heart disease in Aboriginal peoples, Torres Strait Islanders and rural and remote populations. One of the priority areas identified at this workshop was the need for a more coordinated approach to chronic diseases and for the formation of an alliance of non-government organisations (NGOs) to work towards this and to undertake a lobbying and advocacy role. A meeting of a wide range of NGOs working in chronic disease, led by the National Heart Foundation of Australia, was held in Sydney in May 2000. At the Sydney meeting it was agreed that an alliance of NGOs could be formed for the development of a chronic disease strategy for Aboriginal peoples, Torres Strait Islanders and rural and remote populations. The NGOs drafted a 'Statement of Intent', which would inform their work on both heart disease and on broader work to address chronic preventable disease in the target populations. There is a considerable amount of procedural work to be done before the proposed alliance becomes a reality but the prospect of closer collaboration between the NGOs working in chronic disease has much to offer, especially for the population groups that were the focus of the Townsville workshop. This 'alliance' initiative comes at a time when there are national and State/Territory moves on broader aspects of what could become a 'national chronic disease prevention and management strategy'. PMID- 11894294 TI - Measles campaign a success. PMID- 11894295 TI - Sweet solution for easing pain in babies. PMID- 11894296 TI - Nursing near a war zone. PMID- 11894297 TI - Nurse practitioners: defining the achievement. PMID- 11894298 TI - Supporting Koori health workers. PMID- 11894299 TI - Nurses unaware of side effects of psychotropic medications in the aged. PMID- 11894300 TI - The impact of nursing practice on sleep in the older person. PMID- 11894301 TI - Cancer in the elderly a neglected area. PMID- 11894302 TI - Learning a new language. PMID- 11894303 TI - Withholding pain relief is a serious issue. PMID- 11894304 TI - Where is the evidence for rural Medicare abuse? PMID- 11894306 TI - Lethal licorice. AB - The human body's response to extreme stress is complex and this patient's experience illustrates just how traumatic it can be both physiologically and psychologically. While Alan's experience was extreme and unusual, it serves to emphasise the importance of diet and health maintenance particularly for the cardiac patient. PMID- 11894307 TI - AIDS destroys children's lives in Africa. PMID- 11894308 TI - Bev Dark: an isolated and disciplined training. PMID- 11894309 TI - Carolyn Gray: burning the candle at both ends. PMID- 11894310 TI - Maria Beisner: a lifetime of caring. PMID- 11894312 TI - Caesarean section does not guarantee satisfaction. PMID- 11894313 TI - Call to screen pregnant women for HIV. PMID- 11894311 TI - Isabel Walsh: a letter to an old friend. PMID- 11894314 TI - Postnatal depression in aboriginal communities project. PMID- 11894315 TI - Nursing and the law. The facts. PMID- 11894318 TI - How to develop informative, interesting, and organized conference presentations. PMID- 11894317 TI - Gout. PMID- 11894320 TI - The cancer journey: bridging art therapy and museum education. AB - This paper describes the application of art therapy in assisting cancer patients to visually express their cancer experience on several levels--physically, psychosocially, and spiritually. The art therapy/museum education program was developed in 1996 at the McMichael Canadian Art Collection in partnership with Toronto-Sunnybrook Regional Cancer Centre-Bayview Support Network. To date, the program has run for three rounds with 16 sessions in each round. The facilitator for all three rounds was a trained art therapist. The program provided an unique opportunity for an arts institution to serve the community at large by offering an artistic outlet in a peaceful, rural setting in contrast to a sterile hospital environment. The specific goals of the program and the general therapeutic benefits of art therapy are described. In addition, the effectiveness of an existential/phenomenological approach in not only serving the cancer population, but also bridging the two diverse disciplines--art therapy and museum education- is explored. It is suggested that an existential therapeutic approach promotes the confrontation and acceptance of death that is necessary in order to lead a more meaningful life. Moreover, a phenomenological approach promotes the act of "seeing" as an essential ingredient in gaining objectivity and bringing unconscious thoughts into consciousness. The importance of social and emotional support in the way of art therapy in addition to medical care is emphasized. Through art therapy, cancer patients are encouraged to discover ways to face pain and misfortune and be creative in their "art of living." PMID- 11894321 TI - An innovative art therapy program for cancer patients. AB - Art therapy is a healing art intended to integrate physical, emotional, and spiritual care by facilitating creative ways for patients to respond to their cancer experience. A new art therapy program was designed to provide cancer patients with opportunities to learn about the McMichael Canadian Art Collection and to explore personal feelings about their cancer experience through combined gallery and studio components. The role of the facilitator was to assist in the interpretation of a participant's drawing in order to reveal meaning in the art. This paper presents patients' perspectives about the new art therapy program. Content analysis of participant feedback provided information about the structure, process, and outcomes of the program. Evaluation of the art therapy/museum education program demonstrated many benefits for cancer patients including support, psychological strength, and new insights about their cancer experience. PMID- 11894319 TI - Providing care to patients with pancreatic cancer: a retrospective chart review. AB - Pancreatic cancer may be considered rare, yet in Canada it is the fourth leading cause of death by cancer in the elderly. This study was conducted in a large tertiary centre to determine the symptoms experienced by patients and the response by health professionals in providing supportive care. This paper reports the results of a retrospective review of health records from patients diagnosed with pancreatic cancer (n = 99). Results indicate that pain, nausea, vomiting, and anorexia were frequently reported. There was a lack of consistency in the documentation of nursing care and little evidence of an organized, planned approach for care delivery. The role of the interdisciplinary health care team and its members in managing this devastating disease and its impact on patient quality of life was difficult to ascertain. The development of an integrated approach to the care of patients with pancreatic cancer is presented. PMID- 11894325 TI - Get them while they're young. PMID- 11894330 TI - Collaboration is the key to providing continuity of care. PMID- 11894332 TI - Re-evaluating the role of the clinical nurse in minimising health care related infection. PMID- 11894322 TI - Undergraduate recruitment is a major concern. PMID- 11894334 TI - Reducing the risk of SIDS: evidence-based practice. PMID- 11894335 TI - A quality approach to medication management. PMID- 11894336 TI - Vivian Bullwinkel donates diaries to the Australian War Memorial. PMID- 11894333 TI - Medicare undermined by high drug use, intervention and rorting. PMID- 11894337 TI - I.V. fluid therapy. Part 1. Water balance and hydration assessment. PMID- 11894339 TI - Safety worries turn older women off prenatal tests. PMID- 11894338 TI - Campaigning for quality health care. PMID- 11894340 TI - Ending the silence on violence. PMID- 11894341 TI - Give the public system a fair go. PMID- 11894342 TI - Private health incentives should be poured into Medicare. PMID- 11894343 TI - Breast milk is sweet enough. PMID- 11894344 TI - Pressure ulcers--prevention is the key. PMID- 11894345 TI - The perennial pressure sore revisited. PMID- 11894346 TI - A healing success--RDNS Wound Assessment Clinic. PMID- 11894347 TI - Preventing institution--acquired pressure ulcers. PMID- 11894348 TI - Clinical and corporate governance--salvation or just jargon? PMID- 11894350 TI - A new millennium of challenges. PMID- 11894349 TI - Caring for a person with dementia in acute hospitals. AB - Nursing a person with dementia with an acute illness in a hospital setting is a great challenge. This requires negotiating complex interpersonal relationships, and balancing a multiplicity of tasks and roles within social, political and economic constraints. A concerted effort by health professionals and hospital administration to provide appropriate staff education, a multidisciplinary approach, liaison with family carers and access to experts in psychogeriatrics as well as suitable ward environment are positive steps to improving care for a very vulnerable group of patients. PMID- 11894351 TI - Non-nursing duties are eroding our status. PMID- 11894352 TI - Australian nurses are not using wound care research. PMID- 11894353 TI - Risk assessment for pressure ulcers: a comparison of two tools. PMID- 11894354 TI - Getting the sponge count right. PMID- 11894355 TI - I.V. fluid therapy. Part 2. I.V. fluid selection. AB - I.v. fluid selection depends on the estimated fluid loss, the primary fluid compartment involved, the patient's underlying problem and the physiological and haemodynamic impact of the i.v. solution. Clinically, the most important problem is intravascular fluid volume deficit, which is associated with hypotension, inadequate tissue oxygenation and hypoperfusion of essential organs. Intravascular volume resuscitation is therefore of primary importance. Crystalloid solutions have the disadvantage of only small amounts remaining in the IVS whereas colloids are known as plasma volume expanders due to predominantly remaining in the IVS in the presence of an intact capillary endothelium. Managing i.v. fluid administration requires close observation of the patient's subtle responses that may indicate states of fluid depletion or overload. Understanding the physiological principles of the body's fluid distribution in relation to the clinical assessment of the patient's hydration status, together with knowledge of the selected i.v. solution's properties, will enable the nurse to provide quality nursing care and improve patient outcomes. PMID- 11894356 TI - United action for aged care. PMID- 11894360 TI - Aussies nurse East Timor back to health. PMID- 11894361 TI - Aged care under siege. PMID- 11894362 TI - Clinical nursing: the core of our profession. PMID- 11894363 TI - 21st century directions in nurse education at RMIT University. PMID- 11894364 TI - Moral philosopher looks at euthanasia. Interview by Anne Sarzin. PMID- 11894366 TI - Clinical practice is integral to education. PMID- 11894368 TI - Forensic nursing. Formalising a new role or recognising existing practice? PMID- 11894372 TI - Nursing behind razor wire: a question of ethics. PMID- 11894369 TI - Understanding the special needs of the patient with scleroderma. PMID- 11894373 TI - Australia needs more indigenous nurses. PMID- 11894374 TI - The resident classification system: accountability or control? PMID- 11894377 TI - Modern drug treatment options for rheumatoid arthritis. PMID- 11894378 TI - The anxiety of learning. Interview by Diane L. Coutu. AB - Despite all the time, money, and energy that executives pour into corporate change programs, the stark reality is that few companies ever succeed in genuinely reinventing themselves. That's because the people at those companies rarely master the art of transformational learning--that is, eagerly challenging deeply held assumptions about a company's processes and, in response, altering their thoughts and actions. Instead, most people just end up doing the same old things in superficially tweaked ways. Why is transformational learning so hard to achieve? HBR senior editor Diane Coutu explores this question with psychologist and MIT professor Edgar Schein, a world-renowned expert on organizational development. In sharp contrast to the optimistic rhetoric that permeates the debate on corporate learning and change, Schein is cautious about what companies can and cannot accomplish. Corporate culture can change, he says, but this kind of learning takes time, and it isn't fun. Learning is a coercive process, Schein argues, that requires blood, sweat, tears, and a certain level of anxiety to achieve the desired effect. In this article, he describes two basic types of anxiety--learning anxiety and survival anxiety--that drive radical relearning in organizations. Schein's theories spring from his early research on how American prisoners of war in Korea had been brainwashed by their captors. He cites the parallels between the "coercive persuasion" tactics the Chinese communists used to control their prisoners (isolating powerful ones and overseeing all communications) and the corporate boot camps that American companies use to indoctrinate their managers. Indeed, heavy socialization is back in style in U.S. corporations today, Schein says, even if no one is calling it that. PMID- 11894379 TI - Predicting the unpredictable. AB - The collective behavior of people in crowds, markets, and organizations has long been a mystery. Why, for instance, do employee bonuses sometimes lead to decreases in productivity? Why do some products generate tremendous buzz, seemingly out of nowhere, while others languish despite multimillion-dollar marketing campaigns? How could a simple clerical error snowball into a catastrophic loss that bankrupts a financial institution? Traditional approaches like spreadsheet and regression analyses have failed to explain such "emergent phenomena," says Eric Bonabeau, because they work from the top down, trying to apply global equations and frameworks to a particular situation. But the behavior of emergent phenomena, contends Bonabeau, is formed from the bottom up--starting with the local interactions of individuals who alter their actions in response to other participants. Together, the myriad interactions result in a group behavior that can easily elude any top-down analysis. But now, thanks to "agent-based modeling," some companies are finding ways to analyze--and even predict--emergent phenomena. Macy's, for instance, has used the technology to investigate better ways to design its department stores. Hewlett-Packard has run agent-based simulations to anticipate how changes in its hiring strategy would affect its corporate culture. And Societe Generale has used the technology to determine the operational risk of its asset management group. This article discusses emergent phenomena in detail and explains why they have become more prevalent in recent years. In addition to providing real-world examples of companies that have improved their business practices through agent-based modeling, Bonabeau also examines the future of this technology and points to several fields that may be revolutionized by its use. PMID- 11894380 TI - Do you have a well-designed organization? AB - For most companies, organization design is neither a science nor an art; it's an oxymoron. Organizational structures evolve in fits and starts, shaped more by politics than by policies. Although most executives can sense when their organization designs are not working well, few take meaningful action, partly because they lack a practical framework to guide them. The authors of this article provide just such a framework; they present nine tests that can be used to either evaluate an existing organization design or create a new one. Four "fit" tests offer an initial screen: The market advantage test asks whether a design directs sufficient management attention to the company's sources of competitive advantage; the parenting advantage test determines whether the design gives enough attention to the corporate-level activities that provide real value to the company; the people test shows whether the design reflects the employees' strengths; and the feasibility test looks at constraints that may impede implementation. Five "good design" tests can help a company refine its prospective design. The specialist cultures test ensures that there's sufficient insulation for units that need to be different from the prevailing culture; the difficult-links test determines whether a design offers solutions for potentially problematic unit-to-unit links; the redundant-hierarchy test asks whether the design has too many parent levels; the accountability test looks at whether every unit has suitable controls; and the flexibility test ensures that the design lets the company adapt to change. Once a design is altered, the tests should be repeated. Organizational decisions are inevitably complex, and tweaking one part of the design may produce unanticipated consequences elsewhere. PMID- 11894381 TI - The trouble I've seen. AB - David James is a professional crisis manager. For almost 30 years, his job has been to rescue companies on the brink of bankruptcy. By the time he's called in, it's usually too late to save much for the shareholders. In almost every case, however, there is still a lot to salvage: Nearly all the companies James has managed continue to operate in some form. More than 2 billion Pounds have been repaid to banks and unsecured trade creditors. And more than 30,000 jobs have been saved. The key to preserving value, James has found, is to resist the urge to try to generate cash from operating the business. In most cases, these companies have taken on far too much debt to ever sell their way out. Indeed, trying to expand operations beyond what the market would bear was what got them into trouble in the first place. James argues against waiting until the company is dead to break it up. A more effective course is to sell off valuable assets while the company is still a going concern. In vivid and sometimes hair-raising detail, James recounts how he has discovered valuable assets in unexpected places, salvaging everything from airlines to soft-drink makers to Britain's Millennium Dome. He has a routine for accomplishing this, which involves locking up the checkbook on day one and, more often than not, firing the senior management. His is a cautionary tale for top executives who, it is clear, should be concentrating their efforts on never needing to call on him in the first place. PMID- 11894382 TI - Everything I know about business I learned from monopoly. AB - How do game designers approach their work? Perhaps in the same way that managers should. Here, the author, an expert in board-game design and the world's foremost authority on Monopoly, translates six tenets of game design into management principles. Three tenets focus on giving players the right level of structure. First, design simple and unambiguous rules: That also holds true in business; people engage most when responsibilities, objectives, and evaluation criteria are clear. Second, avoid frustrating the casual player. Just as not every game player aspires to be a grand master, not every employee wants to think like an executive. Third, establish a rhythm so that players know intuitively whether they are at the beginning, middle, or end of the game. Managers can also engineer such shifts of momentum and motivation for workers. Three more principles focus on providing entertainment. The most important is to tune into what's happening off the board. For many people, the real joy of a great game--or a great job- comes from the larger social experience surrounding it. Another key is to offer chances to come from behind. Even struggling employees want to believe, "The odds may be stacked against me, but just one great stroke and I'm right back in it." Finally, managers, like game designers, should provide outlets for latent talents. Games themselves can be useful in the workplace. For instance, an afternoon of game playing builds relationships and increases an organization's social capital. And simulation games can sharpen employees' business judgment. Managers may come to appreciate that games succeed depending on how well designed they are--and that many design challenges have their equivalents in the art of management. PMID- 11894383 TI - The 2002 HBR list. Breakthrough ideas for today's business agenda. AB - In the last year, war broke out, the economy took a nosedive, and an alarming number of businesses went belly up. Thus it is with some urgency that we bring you this year's list of the seven best new business ideas to help you find your way through these complex times. History Returns. Many thought that the fall of the Berlin Wall marked the beginning of a new world order, one in which history didn't matter. But September 11, 2001, put an end to that theory. And it raised serious questions about globalization, security, and strategy. Enter the Everyday Leader--at Last. CEOs are used to getting all the glory, but leaders outside the limelight--middle managers and tempered radicals--are beginning to receive the attention they deserve. The Internet Is Not About You. The real value of the Internet may lie less in connecting individuals than in connecting databases, servers, and devices--a web not of people but of machines. Mind Your Behavior. Behavioral scientists are beginning to be able to accurately predict the ways individuals and crowds will respond to stimuli, and the implications for business are profound. Don't Delight Your Customers Away. Companies have wooed and coddled customers for too long. The truth is, people delight in being teased and are repelled by those who try too hard to befriend them. Games Are for Losers. Financial game playing and verbal politicking are seriously damaging businesses. Honesty is more than an admirable virtue; it's the foundation of every lasting enterprise. Three Cheers for Creativity (Sometimes). Creativity and best-practice replication are fundamentally different undertakings. Managers who mix the two achieve creativity that is merely incremental and replication that is sadly inept. PMID- 11894384 TI - The virtue matrix. Calculating the return on corporate responsibility. AB - Executives who want to make their organizations better corporate citizens face many obstacles: If they undertake costly initiatives that their rivals don't embrace, they risk eroding their company's competitive position. If they invite government oversight, they may be hampered by costly regulations. And if they adopt wage scales and working conditions that prevail in the wealthiest democracies, they may drive jobs to countries with less stringent standards. Such dilemmas call for clear, hard thinking. To aid in that undertaking, Roger Martin introduces the virtue matrix--a tool to help executives analyze corporate responsibility by viewing it as a product or service. The author uses real-life examples to explore the forms and degrees of corporate virtue. He cites Aaron Feuerstein, CEO of Malden Mills, a textile company whose plant was destroyed by fire in 1995. Rather than move operations to a lower-wage region, Feuerstein continued to pay his idled workforce and rebuilt the plant. Unlike the typical CEO of a publicly held corporation, who is accountable to hundreds or thousands of shareholders, Feuerstein was free to act so generously because he had only a few family members to answer to. But as Martin points out, corporations don't operate in a universe composed solely of shareholders. They can be subject to pressure from citizens, employees, and political authorities. The virtue matrix provides a way to assess these forces and how they interact. Martin uses it to examine why the public clamor for more responsible corporate conduct never seems to abate. Another issue the author confronts is anxiety over globalization. Finally, Martin applies the virtue matrix to two crucial questions: What are the barriers to increasing the supply of corporate virtue? And what can companies do to remove those barriers? PMID- 11894385 TI - The hidden challenge of cross-border negotiations. AB - Cultural differences can influence business negotiations in unexpected ways, as many a hapless deal maker has learned. But the differences extend well beyond surface behaviors, such as proper table manners and the exchange of business cards--and even beyond deeper cultural characteristics, such as attitudes about relationships and deadlines. Indeed, there's another, equally treacherous aspect to cross-border negotiation: the ways that people from different regions come to agreement, or the processes involved in negotiations. Decision-making and governance processes can vary widely from culture to culture, not only in terms of legal technicalities but also in terms of the behaviors and core beliefs that drive them. Numerous promising deals have failed because people ignored or underestimated the powerful differences in process across cultures. In this article, James Sebenius offers ways in which negotiators can prepare for such cultural differences. A useful approach, he says, is to map out the decision making process--including who's involved, what formal and informal roles people play, and how a resolution is actually reached. With that knowledge, you can design a strategy that anticipates obstacles before they arise. Governance and decision-making processes can take devilishly unexpected forms as you cross borders. But by designing your strategy and tactics so that you're reaching all the right people, you increase your chances of striking a sustainable deal. Those negotiations that might otherwise have failed because people ignored or underestimated powerful disparities in process will, in the end, yield a meaningful yes. PMID- 11894386 TI - Making sense of corporate venture capital. AB - Large companies have long sensed the potential value of investing in external start-ups, but more often than not, they fail to get it right. Remember the dash to invest in new ventures in the late 1990s and the hasty retreat when the economy turned? This article presents a framework that will help a company decide whether it should invest in a particular start-up by first understanding what kind of benefit might be realized from the investment. The framework--illustrated with examples from Intel, Lucent, and others--explains why certain types of corporate VC investments proliferate only when financial returns are high, why other types persist in good times and in bad, and why still others make little sense in any phase of the business cycle. The framework describes four types of corporate VC investments, each defined by its primary goal--strategic and financial--and by the degree of operational linkage between the start-up and the investing company. Driving investments are characterized by a strong strategic rationale and tight operational links. Enabling investments are also made primarily for strategic reasons, but the operational links are loose. Emergent investments, which are characterized by tight operational links, have little current--but significant potential--strategic value. Passive investments, offering few potential strategic benefits and only loose operational links, are made primarily for financial reasons. Passive corporate VC investments dry up in a down economy, but enabling and driving investments usually have more staying power. That's because their potential returns are primarily strategic, not financial. In other words, they can foster business growth. Emergent investments may make sense even in a weak market because of their potential strategic value- that is, their ability to help companies identify and spark the growth of future businesses. PMID- 11894387 TI - [Telepathology--applications of an emerging technology]. AB - Due to the development of modern telecommunication by means of high speed data transfer either by normal telephone lines or the internet we are faced with new possibilities and chances in pathology. Instead of mailing slides, today images can be transferred electronically within seconds around the world, a second opinion of a highly skilled specialist can be available within the same day or even the same hour. Beside teleconsultation, telediagnostics is the most sophisticated application of telepathology. This means the evaluation of freshly prepared cryosections obtained during surgery by a pathologist using a remotely controlled microscope in a location deprived of pathological expertise. Since the technology applied has become very reliable, problems encountered by using this technology are mainly caused by either microscopic or macroscopic sampling errors. A correct diagnosis can not be obtained if the images mailed are not containing the necessary information. The same is true for telediagnostics, if the biopsy taken for cryosection does not contain the relevant lesion, a correct diagnosis will not be possible. However, these problems are not specific for telepathology, but are encountered in routinely processed specimens as well. Thus, protocols need to be developed assuring sufficient and reproducible image sampling for teleconsultation. PMID- 11894388 TI - [Unnatural causes of death--what the pathologist must know]. AB - The lecture explains the current legal position in Germany and is intended to help pathologists make decisions. I. The term "unnatural death" is a technical term within the meaning of section 159 German Code of Criminal Procedure. Even "indications"--again a legal term--constitute a statutory duty to make an immediate report. If such causes for suspicion arise at autopsy, the pathologist is confronted with an ethical and legal dilemma. With his report he can trigger a chain of events that is no longer under his control, and may extend well beyond the original request for a medical diagnosis. II. Starting with the relevant definitions for the subject, the lecturer focuses on the special case of unexpected death in connection with medical intervention. Where does the pathologist's responsibility end? Which information channels must be observed? The pathologist's rights, duties and degree of discretion are discussed. III. Finally, the lecturer addresses the "matter of the body" which must be examined by the responsible state prosecutor as a consequence of the pathologist's report. PMID- 11894390 TI - [Atherosclerosis--search for a new entity]. AB - Atherogenesis is a disease of middle-sized and large caliber blood vessels that can be divided into three major phases. The initial lesions of early atherosclerosis are characterized by the adhesion and subendothelial emigration of blood-borne monocytes, which differentiate into macrophages and provide the morphological basis for the formation of foam cells and fatty streak lesions. These lesions are found in most children and teenagers in industrialized nations. The next key event in atherogenesis is the proliferation of smooth muscle cells within the intima and media, resulting in the gradual compromise of the vessel lumen. Myofibroblastic cells also contribute to lesion growth through the production of excessive amounts of extracellular matrix. Such lesions are clinically silent unless progression to the next phase continues: the lesions degenerate, forming a mostly necrotic "lipid core" consisting of extracellular lipid, cholesterol crystals, inflammatory cells and necrotic debris. A fibrous cap is formed which prevents the interaction of blood cells, particularly of platelets with the highly proaggregatory material found in the lipid core. However, continuous inflammatory activity and/or heightened mechanical stress (i.e. in hypertension) tends to weaken the fibrous caps. Eventually, plaque rupture ensues, platelets aggregate, and the lesions become clinically manifest in such dramatic events as myocardial infarction, stroke, or mesenteric ischemia. Research into lesion formation and progression is limited by the fact that lesions develop in silence over many decades and that animal models only incompletely model the situation in humans. Most currently debated concepts accept the "response to injury" hypothesis formulated by the late Russell Ross and the multifactorial nature of atherogenesis. The discussion today circles around the relative contributions of low density lipoproteins (oxidized or enzymatically modified LDL?), the immune response (adaptive or innate?), infectious agents (CMV, chlamydia pneumoniae?), and/or hereditary factors, to name only a few of the most widely debated concepts. Irrespective of the outcome of this pathomechanistic discussion, the knowledge of established risk factors (hypercholesterolemia, hypertension, diabetes, smoking, etc.) and protective interventions (lifestyle changes, physical exercise, "healthy" diets, effective dietary and pharmacological control of hyperglycemia, blood pressure or hyperlipidemia) has helped to define atherosclerosis as a "new entity" that has little to do with the archaic concept of a "degenerative" vessel disease. PMID- 11894389 TI - [The significance of the autopsy--now and in the future]. AB - The significance of the autopsy is reflected in the quality control of medical diagnosis and therapy, as well as in the social relevance of the autopsy findings for the individual, family and society in general ("individual" and social epidemiology). Contemporary statistics from research of the literature and internet will be presented and interpreted in a modern context. A pre-requisite for a quality control function is an adequately high autopsy rate, which should be a factor taken into consideration for the registration und certification of a clinic or hospital. There are some valuable studies cited which assess the significance of autopsy as a means of quality control in clinical surgery, oncology and general internal medicine. There is, on the other hand, a decline of autopsy rates worldwide, but especially in the Federal Republic of Germany since the beginning of the 90ties. In addition, the causes will be demonstrated and discussed briefly. The attitude of medical colleagues and health insurance companies, as well as public opinion and politics, all influence the significance of and attitude to the autopsy, including the readiness to create and maintain a transparent legal and cost-covering framework. A spectrum of the differing opinions, obtained from various discussions, will be given. In addition, the necessary legal and financial regulations to facilitate and sustain the autopsy service at a sufficiently high rate (at least 30%, based on statistical considerations) will be discussed in the context of desirable short- and long term solutions. PMID- 11894391 TI - [Vasculitis--aspect of cellular and molecular pathogenesis]. AB - Vasculitis, inflammation of the vessel wall, can be observed in acute and chronic inflammatory processes, in vascular rejection of allogeneic transplants and can be encountered as primary vasculitis of arterial, arteriolar, capillary and venular vessels. Although the numerous forms of vasculitis and their associations with different diseases result in a multitude of etiologic and pathogenetic factors there are pathogenetic factors common to several vasculitides. These include innate immunity factors, transcription factors such as NFkB, endothelial cytoprotective agents such as NO and anti-neutrophilic cytoplasmic auto antibodies (ANCA). ANCA may be directed against several antigens, in the majority of cases against proteinase 3 and myeloperoxidase. The complex of proteinase 3 and ANCA leads to an increased expression of CD18, CD14 and an elevated synthesis of cytokines and chemokines such as interleukin 1, interleukin 8 in monocytes. In granulocytes generation of reactive oxygen species is found in addition. In both cells apoptosis finally occurs. ANCA may also bind to a surface glycoprotein (gp130) expressed on glomerular and peritubular endothelia in the kidney. Thus the activation of granulocytes, monocytes and endothelial cells by ANCA may be a critical step in the initiation phases of vasculitis. NO is cytoprotective for endothelial cells in small concentrations. Our group has shown in detailed studies that inhibition of endothelial NO synthase is detrimental and enhancement of activity of endothelial NO synthase is beneficial for allogeneic solid organ transplants. The transcription factor complex NFkB is a key regulatory transcription factor for the expression of genes and proteins associated with acute inflammatory processes and endothelialitis. Inhibition of NFkB activity by a decoy-oligonucleotide prevented activation of endothelial calls in reperfusion injury and vascular rejection. The complement system probably plays an essential role in the initiation and propagation phases of vasculitis. Specifically the pneumococcal C-polysaccharide-reactive protein (CRP), synthesized after trauma and infection, can potently activate the complement cascade which leads to an activation of endothelial cells with increased expression of adhesion molecules. The 4 shortly described pathogenetic mechanisms of vasculitis seem to be important and common factors for the generation and maintenance of vascular inflammation; nevertheless these factors are only part of the spectrum of different humoral and cellular responses in vasculitis. The described experimental investigations on endothelial damage and endothelialitis may lead to new therapeutic strategies in vasculitis. PMID- 11894392 TI - [Phenotype--genotype--correlation in ovarian neoplasia]. AB - Ovarian neoplasms display a wide range of phenotypic differentiation patterns. In the recent past, molecular genetic aberrations have been increasingly identified in various types of ovarian tumors. Granulosa cell tumors most often contain numeric chromosomal aberrations (monosomy 22, trisomy 12 and 14). Numeric changes can also be found in benign and borderline epithelial neoplasms, however without demonstrating specific patterns. K-ras mutations are characteristic for mucinous ovarian tumors and for serous borderline (LMP) tumors. In serous LMP tumors they are associated with low level microsatellite instability. Complex chromosomal aberrations are not detected in benign and borderline tumors. Invasive ovarian carcinomas show complex genetic changes. Chromosomal gains at 3q26, 8q24 and 20q13 apparently represent early lesions, whereas loss of material of chromosomes 4, 13, 16, 18 and X is associated with tumor progression and poor prognosis. The main targets of chromosomal changes are regulatory genes of cell proliferation and apoptosis (e.g. p16, cyclin D1, Rb, p53, myc, bcl-2) and members of the signaling cascade of tyrosine kinase receptors (e.g. Her-2/neu, dab-2, K-ras, PI3 K, PTEN). The genetic alterations of ovarian neoplasms described so far apparently correlate with the different level of aggressiveness. However, they do not fully explain the spectrum of phenotypic variability of these tumors. PMID- 11894393 TI - Anti-CD20 treatments and the lymphocyte membrane: pathology for therapy. AB - Therapeutic antibodies directed against the CD20 surface protein of B-lymphocytes are efficient means of controlling the growth and survival of malignant B lymphocytes. The mechanisms of anti-CD20 antibody action remain in great part unexplained. However, we show that incubation of CD20+ cells with therapeutic antibodies such as Rituximab results in the redistribution of the CD20 marker in plasma membrane rafts. Rafts are specialized membrane organizations of sphingolipids and cholesterol in the plasma membrane outer leaflet that serve as signalling platforms in lymphocytes and other cells, and allow transmembrane propagation of most receptor-mediated extracellular signals. This redistribution of CD20 to rafts is not acutely toxic to lymphoma cells, but leads to long lasting perturbations of transmembrane signalling and contributes to the progressive elimination of B-lymphoma cells. The accumulation of CD20 in rafts causes downmodulation of raft-associated protein tyrosine kinases and modifies the spatial relationships between raft lipid and protein components. Such modifications of the raft structure and function are likely to cause relatively long-lasting perturbations of the lymphoma cell physiology and contribute to the elimination of the Rituximab-targeted cells. PMID- 11894394 TI - [Multimedia fetal pathology]. AB - Due to the enormous technical developments in recent years in the area of prenatal ultrasound more malformations are detected in greater detail and at an earlier point in time during pregnancy than in the past. As a result, fetuses of increasingly smaller size and with complex diagnoses are presented for autopsy. At the same time, the genetic causes of ever more diseases, malformations and syndromes are being discovered, so that DNA diagnostics, chromosome analysis and, among others, molecular-genetic and biochemical investigations have come increasingly to complement clinical-genetic diagnostics. The increasing clinical requirements also raise the requirements for advanced training in the area of fetal pathology. Multimedia techniques can help satisfy these requirements. Within the framework of routine diagnostics representative findings in exemplary cases can be worked out multimedially. These cases could be presented by CD or shown on the World Wide Web. Tele-pathology can come to play an essential role in advanced training: seminars can be conducted directly or virtually using preserved specimens or direct expert consultations can be conducted for the increasingly complex routine cases. The technical development mentioned above makes it clear that advanced training has to be carried out within a clinical context. Commonly accessible interdisciplinary databases play a central role here. PMID- 11894395 TI - [pathology of the umbilical cord in relation to gestational age:. Findings in 4,267 fetal and neonatal autopsies]. AB - The umbilical cord is the "foetal lifeline". Pathological changes of the umbilical cord may strongly threat foetal life. Otherwise there are many morphological abnormalities without any influence on foetal well-being. There is poor knowledge about many special pathologic-anatomical features of the cord. AIMS: Our aim was to study the incidence and relevance of pathological changes of the umbilical cord, including so-called "cord accidents", in our large autopsy series. METHODS: Our study is a review of 4267 cases, including 1301 aborted foetuses and 2966 stillborns and lifeborns less than 7 days of age, autopsied from 1971 to 1996 in our department. RESULTS: Of 1301 aborted foetuses, 94 (7.2%) had abnormalities of the umbilical cord. Most of them (about 59%) showed torsion, stricture or "Thin-cord-Syndrome". Other important lesions of the cord in aborted foetuses are omphaloceles and constrictions of the cord secondary to Streeter bands. Most of all changes of the cord found in aborted foetuses were lethal factors. Of 2966 stillborn and lifeborn children, which died before day 7 of life, 456 (15.4%) had abnormal findings in relation to umbilical cord. 228 children showed umbilical cord accidents (i.e. true knots, cord prolapse, cord entanglement). Likewise in 228 children, we found especially changes of cord vessels like singular umbilical artery and others, cord oedema, insertion anomalies and other changes. This findings often represent no lethal factors. CONCLUSIONS: Torsion, stricture, and the complex of the thin umbilical cord play the most important role especially in abortion, but is also the most important cause of death of the older foetus in relation to true umbilical cord pathology. PMID- 11894396 TI - [Pathology in Munster]. PMID- 11894397 TI - [Histopathologic breast diagnosis in view of consultant studies of the Mamma Regisstry Fulda]. AB - The Mamma-Registry Fulda, founded in 1976 by the senior co-author, is a personal service for pathologists having problems with histological diagnosis of breast diseases. From nearly 7000 cases filed up to now we selected 1112 consecutive consultations from 1996 to 1999 for this study. The aims were a critical analysis of a "submission-profile" and for each case a comparison of submitters' diagnoses with that of the register to crystallize special fields of problems in histopathological diagnosis and to make a statement about quality standards which was shown in a raster of results. The submitted cases came from pathologists in university institutes (13.9%), city hospitals (49.0%), group practices (24.2%), and single practices (11.6%). The material consisted of selected paraffin-blocs in about two thirds and of slides only in less than one third. The sendings were accompanied by letters with sufficient information on history, clinical background, and gross findings in 72%, and in an additional rate of 17.1% by copies of the histological reports already given by the submitters to their clinicians. The main reasons for consultations were a primarily uncertain diagnosis (45.8%) or the request to affirm a more or less definite diagnosis (40.7%) in cases of rare lesions or differing judgements in the submitting institution. Each diagnosis of the registry was coded in a special diagnostic key. In a raster of results the diagnosis of each case was listed as identical (55.0%) when there was complete agreement between submitter's and register's result, as included (23.9%) when one of the differential diagnoses named by the submitter fitted the register's diagnosis, and as different (6.7%) when there was only agreement about the dignity of differently classified lesions. False positive (2.3%) and false negative (4.5%) diagnoses of submitters were subclassified as clinically irrelevant (2.6%) and relevant (4.2%). The most often missed diagnostic entity in the latter group was tubular carcinoma. Overall the results of the study justify awarding a high standard of quality to histopathological diagnostics. PMID- 11894398 TI - [Predictive and prognostic factors in breast cancer evaluated by immunohistological methods]. PMID- 11894400 TI - [Esophageal, gastric and colorectal tumors]. AB - The new WHO-classification of the tumours of the digestive System replaces the "blue books", and are now dealt with completely in a single book. In addition to the histological features of the lesions, the new classification also contains information on epidemiology, aetiology, endoscopy, genetic susceptibility, molecular genetics, prognosis and predictive factors. The erstwhile mostly black and-white histological photographs have at last again been replaced by numerous colour photographs and supplemented by endoscopic and macroscopic pictures. The changes to the individual tumour classifications are only few. The former classification of the malignant lymphomas of the gastrointestinal tract has now been replaced by the classification of these lesions that has long been in use. New additions are the gastrointestinal tract has long been in use. New additions are the gastrointestinal stroma tumours (GIST) and the gastrointestinal autonomic nerve tumour (GANT). To the epithelial tumours of the oesophagus have now been added the basaloid squamous cell tumours of the vermiform appendix and the colorectum have now been added the serrated adenoma and the small-cell carcinoma. The following new chapter have been included: adenocarcinoma of the oesophagogastric junction, secondary carcinomas of the stomach, secondary tumours of the small bowel and colon, the Peutz-Jeghers syndrome, juvenile polyposis, familial adenomatosis coli, and HNPCC. For the first time the intra-epithelial neoplasias in chronic inflammatory bowel disease have been differentiated, i.e. adenomas distinguished from the dysplasias, while the latter term has now been replaced by the term "intra-epithelial neoplasias". In comparison with the former "blue books", the new WHO-classification, prepared by presentatives of numerous disciplines--for the first time including clinicians--has taken on the character of a text book. PMID- 11894399 TI - [Keratin 5-positive cells in the breast are progenitor cells of glandular and myoepithelial differentiation. A cell biological model as a basis of breast pathology]. AB - In the present computer-assisted labelling study we demonstrate for the first time that in normal breast tissue keratin (K) 5+ progenitor cells give rise to either glandular (K 8/18+) or to myoepithelial cells (sm-actin = SMA+) via intermediary cells (either K 5/6+; K 8/18+ or K 5/6+; SMA+). Furthermore we have conclusive evidence that benign proliferative breast disease lesions are progenitor cell-derived lesions. In contrast, most breast cancers appear to evolve from glandularly differentiated cells. Western blotting experiments which show large amounts of K 5 in benign proliferative breast lesions, but not in ductal carcinoma in situ confirm the immunofluorescence data. Our results pave the way towards a new cell biological model of breast regeneration, benign proliferative breast disease and breast cancer. PMID- 11894402 TI - WHO-classification 2000: exocrine pancreatic tumors. AB - The new WHO-classification is more than a histological description of the tumors of the exocrine pancreas. It is a clinicopathological overview that also covers the most recent genetic data on each tumor-type. New issues concern the possible ductal adenocarcinoma precursor lesions and the further characterization of pancreatic neoplasms showing a ductal phenotype. Pancreatic duct lesions have been discussed as tumor precursors. We will present the recently adopted standard system for these lesions, which distinguishes between three grades of pancreatic intraepithelial neoplasia (PanIN). Molecular studies revealed that PanIN-2 and PanIN-3 lesions represent a distinct step towards invasive carcinoma. Another focus of the review will be the advances that have been made in the further immunocytochemical and molecular characterization of special pancreatic neoplasms showing a ductal phenotype, such as undifferentiated carcinoma, mucinous noncystic (colloid) carcinoma, intraductal papillary mucinous neoplasm, mucinous cystic neoplasm, medullary carcinoma and other rare tumors. PMID- 11894401 TI - [The new WHO classification of liver tumors--what is really new?]. AB - The new WHO (2000) Classification comprises all gastrointestinal tumours including tumours of the liver and intrahepatic bile ducts. The differences between the old edition from 1994 and the new classification will be outlined. A comparison was made with regard to new entities, definitions and tumor subtypes. The eight tumour subgroups of the 1994 classification have been reduced to seven in the 2000 WHO edition, omitting the "unclassified tumors" and changing the group's terms. The 2000 classification shows some minor changes in the ICD-O numbers and in terminology. An important change and hopefully beneficial progress has been achieved by introducing more detailed definitions of epithelial abnormalities, namely liver cell dysplasia and its different subtypes (large cell type dysplasia, small cell type dysplasia, dysplastic nodules low-grade and high grade). The miscellaneous lesions (former called tumour-like lesions) have been reduced in number. In contrast to the 1994 classification which only contained definitions and short descriptions of macroscopic and microscopic findings as well as only sparse clinical information, the 2000 classification offers much more text with new information on epidemiology, aetiology, clinical features, spread, macroscopy, microscopy, genetic susceptibility, and genetics. Some minor differences between the old and new WHO classification can be recognized. Major changes comprise the extended terminology in precancerous epithelial changes, e.g., liver cell dysplasia and reduction of the amount of text. PMID- 11894403 TI - [The man in the ice under special consideration of paleo-pathological evidence]. AB - Dealing with archaeological findings, the physical condition of the 5000-year-old Man in the Ice will be discussed. Several research projects with autoscopical, radiological, computertomographical and laparoscopical methods present an overview of the state of health of the Tyrolean Iceman. The vascular network at the lower brain shows a slight hardening of the arteries. On the right side of the thorax the sixth and seventh ribs are fractured without callus formation. On the left side serial rib fractures can be seen which had healed well. The twelfth ribs are absent on both sides. The Iceman was suffering from arthrosis of the right hip of medium severity. Osteolysis of the little toe is indicative of frostbite. Some ecto- and endoparasites were found (Pulex irritans, Trichuris trichiura). The Iceman has a number of skin markings on various places. The location of these tattoos matches the X-ray findings of discrete to medium arthrotic changes in the corresponding joints. It seems reasonable to assume that the markings were applied for a therapeutic reason. The combination of thickening of the arteries, arthrosis of the joints, the healed fractures and frostbite lesions illustrates the hard life to be endured by the Man in the Ice and by Late Stone Age man in general. PMID- 11894404 TI - [Medicine and pathology in the future]. PMID- 11894405 TI - [The Rudolf Virchow Prize 2001. The role of the oncoprotein beta-catenin ni the progression of colorectal cancers]. AB - Invasion and dissemination of well-differentiated carcinomas are often associated with loss of epithelial differentiation and gain of mesenchymal-like capabilities of dedifferentiated tumor cells at the invasive front. However when analysing central areas of metastases of colorectal carcinomas one finds a regain of the differentiated epithelial growth patterns like in the primary tumor. More than 80% of these tumor have loss of function mutations in the APC tumor suppressor gene, leading to an overexpression of beta-catenine. In its nuclear pool beta catenine acts as a transcription factor and is now considered as one of the main oncogenic proteins in colorectal carcinogenesis. We could define several molecules important for the processes of invasion and dissemination, like MMP-7, uPA, laminin-5, as target genes activated by nuclear beta-catenine. Moreover the characteristic phenotypic changes during tumor progression were associated with distinct expression patterns of beta-catenine and E-cadherin. Nuclear beta catenine was found in dedifferentiated mesenchyme-like tumor cells at the invasive front, but strikingly, like in central areas of the primary tumors, was localized to the membrane and cytoplasm in polarized epithelial tumor cells in the metastases. This was accompanied by changes in the proliferative activity. Based on these data, we postulate that an important driving force for progression of well-differentiated colorectal carcinomas is the specific environment, initiating two transient phenotypic transition processes by modulating intracellular beta-catenine distribution in the tumor cells. PMID- 11894406 TI - [Role of podocyte damage in the pathogenesis of glomerulosclerosis and tubulointerstitial lesions: findings in the growth hormone transgenic mouse model of progressive nephropathy]. AB - The sequence of structural changes terminating in glomerulosclerosis, tubular atrophy and interstitial fibrosis was analyzed in the growth hormone (GH) transgenic mouse (TM) model of progressive renal disease. The investigation was performed in TM expressing the bovine GH gene under the control of the murine metallothionein-1-promoter and non-transgenic controls (CM) of different age groups. The kidneys were studied by light microscopy, transmission and scanning electron microscopy, and were analyzed with stereological methods. Early-stage renal lesions were characterized by glomerular hypertrophy and mesangial expansion. In 7-week-old TM the mean glomerular volume was twice that of age matched CM. The number of endothelial and of mesangial cells per glomerulus was increased in TM vs. CM, while the number of podocytes did not change. The podocytes demonstrated hypertrophy and foot process effacement. Concomitant with an age-related further increase of glomerular size in TM, severe maladaptive podocyte lesions including detachment of podocytes were observed. The resultant denudation of the glomerular basement membrane was associated with severe proteinuria, glomerular hyalinosis, synechia formation and collapse of glomerular capillaries. These lesions progressed to glomerular obsolescence that was associated with atrophy of the adjacent tubule and interstitial fibrosis. The progressive kidney lesions in this model appear to be attributable to a considerable extent to podocyte damage resulting from the limited capacity of this cell type to keep up with progressing overall tuft growth. The findings provide further evidence that mature podocytes are unable for effective cell replication in vivo, and that podocyte damage plays a significant role in the pathogenesis of progressive glomerulosclerosis with tubular atrophy and interstitial fibrosis. PMID- 11894407 TI - [Oncogene amplification and genetic heterogeneity in the metaplasia-dysplasia adenocarcinoma sequence of Barrett esophagus]. AB - AIMS: Information about numerical genomic alterations in the tumorigenesis of Barrett's adenocarcinoma (BCA) is still limited. In order to search for gene amplification and ploidy status, a series of locus-specific DNA probes and associated centromere probes was analysed in the metaplasia-dysplasia adenocarcinoma-sequence. METHODS: Fluorescence in situ hybridisation (FISH) was performed on paraffin sections with locus-specific DNA probes for D7S486, c-myc, cyclin D1, Her-2/neu, 20q13.2 and associated chromosomes 7, 8, 11, 17 and 20. Corresponding areas of intestinal metaplasia (IM, n = 5), low grade dysplasia (LGD, n = 9), high grade dysplasia (HGD, n = 15) and BCA (n = 16) were analysed. RESULTS: Gene amplification of c-myc, Cyclin D1, Her-2/neu and 20q13.2 was observed in 15-35% of BCA. Coincident amplification of genes was also present. Polysomies for all investigated centromere probes were highly prevalent (up to 85%). Gene amplification was also demonstrated in HGD lesions. Polysomies were observed in HGD in high frequency (up to 80%). Extensive genetic heterogeneity was observed in both, BCA and HGD displaying different levels of amplification. None of the samples with LGD showed a locus-specific amplification, but polysomies for all investigated chromosomes were present in 18-48% of LGD. No changes were detected in BCA associated IM and squamous epithelium. CONCLUSIONS: Our data indicate that oncogene amplification of c-myc, cyclin D1, Her-2/neu, and 20q13.2 occurring in BCA and less frequently in HGD is a late event in the tumorigenesis. Polysomies of chromosomes 7, 8, 11, 17 and 20, which were highly prevalent in BCA and HGD occur already at the stage of LGD. This may be a result of an early polyploidization, preceding the later genetic events, such as gene amplification in HGD and BCA. The detection of shared numerical genomic changes and the detected extensive genetic heterogeneity in the metaplasia-dysplasia carcinoma-sequence in Barrett's esophagus supports the hypothesis of a process of multiclonal expansion underlying this progression. PMID- 11894408 TI - [Promoter hypermethylation during tumor progression: quantitative analysis employing kinetic PCR]. AB - The methylation of cytosine residues in the promoter region of tumour-suppressor genes is now widely recognised as an alternative mechanism of gene inactivation in cancer. To address the question whether promoter methylation is an early event in the process of malignant transformation and whether quantitative changes are occurring during clonal evolution we developed a new assay for the measurement of cytosine methylation, which could also be applied to laser-microdissected archival specimens. We first improved the bisulfite conversion of genomic DNA in order to analyse cells isolated from archival tissue sections by laser-assisted microdissection. Analysing the methylation of key regulatory genes in archival specimens (n = 38) we could demonstrate that aberrant promoter hypermethylation is an early event during breast cancer development and changes quantitatively and in a gene-specific manner during progression. This novel methodological approach allows the morphology-guided quantitative analysis of changes in the methylation pattern during the clonal evolution of tumour cells. Quantitative differences detected by this assay will offer new insights into early steps in carcinogenesis and tumour cell heterogeneity. PMID- 11894409 TI - [Identification of new substrates of cyclin-activated kinases in breast cancers]. AB - Mitogenic and growth-inhibitory signals influence cell-cycle progression through their action on a family of cyclin-dependent kinases (cdks). The activity of cdk complexes is regulated in part by the association of a cyclin partner that acts as a positive effector. These cyclin/CDK complexes promote cell cycle progression by the phosphorylation of key substrates. Cyclin D1 and E are frequently overexpressed in breast cancers and cyclin E overexpression has been correlated with a poor prognostic outcome. The in vivo substrates of cyclin E/CDK2 are, however, not well defined. We screened for cyclin E/CDK2 substrates in nuclear extracts of breast cancer cells as well as using a spotted-array protein library with purified active cyclin E/CDK2 complexes that were expressed in and purified from insect cells. Using this technique several potential cyclin E/CDK2 substrates were isolated. Further work is presently underway to identify these substrates and verify their authenticity as in vivo substrates of cyclin E/CDK2. These potential new substrates will help to unravel the highly complex mechanisms of cell cycle control and perhaps offer new targets for the diagnosis and/or treatment of breast cancer patients. PMID- 11894410 TI - [Organization and contents of future research in pathology]. AB - The organisation of research in Pathology in the future will be driven by the pressure for profitableness. This situation is already now leading to a continuous shifting of capacity from "non profitable" research (and teaching) to diagnostics. Since research continues to be a central future goal of the university institutes of Pathology, we have to claim that all our institutes shall have a financially incontestable research unit (at least 1 x BAT1 and 1 x BAT2, 1-2 BATV and consumables for approx. TDM75/year) and appropriate space. Additionally, academically orientated full pathologists and fellows, research assistants on grant money and M.D. students will perform research. One of the central future goals must be increase in motivation of researchers through output orientated individual support and the enhancement of their career expectations. Research planned by politicians ("planned research") can--as "planned economy"- not be successful. Main goal of our future research efforts in the postgenomic era will be further clarification of the molecular basis of diseases. Applied research will be driven by the need for molecular classifications including individual prediction of prognosis, progression and choice and response to therapy. The future face of Pathology entirely depends on the fact, how actively this type of research is done in our labs and, consequently, how well we will be trained to do non-morphologic diagnostics on tissue in the future. PMID- 11894411 TI - [Teaching pathology in the 21st Century]. AB - Medical school teaching undergoes major changes in predictable cycles. Up until the present day, all previous reforms have focused on the improvement of various teaching aspects. Future reforms, however, will be different since they will be based on a different philosophy. That philosophy will be one in which there will no longer be the "teaching aspect" at the center of attention but the "learning aspect". Students will better be prepared how to learn during their medical school education and throughout their entire careers. Worldwide, various models to reform teaching and learning in medical school have been tried. Problem oriented teaching has gained considerable interest. The recently reformed medical school curriculum in Basel focuses on tutorial based problem-oriented teaching and hands-on "problem-oriented learning" in private practice groups as well as various practical courses. Our 3-year experience with this new type of curriculum is very positive. However, at present it is too early to know whether our positive "initial" impression is reflected in improved examinations scores. Experience made in other institutions suggests that such a new type of curriculum is not necessarily accompanied by better test results. Only time will tell. A final critical evaluation of our new "problem-oriented" curriculum can only be made after many years. In case we succeed and students grasp the concept that life-long learning is not only a necessary obligation in medicine but also a natural part of life, all of our efforts will be worthwhile. PMID- 11894413 TI - [Is disease grounded in the mind, the spirit or communication? Pathology as a leading path in learning about disease]. AB - What is the reason, why pathology could command a dominating position as a central teaching discipline in the last 150 years? Furthermore: can this position be maintained in the future? Pathology takes its definitive stand in the philosophic confrontation of "Mind" versus "Matter". It guards the dogma of Morgagni: every disease is reflected by its typical morphological substrate. As long as this dogma is guarded and defended by the discipline of pathology, we will stay at the helm of teaching in medicine. Recently information as a third reality (besides mind and matter) has come into existence. Information is defined here as genomic information specifically the differentially activated functional genomics. In extension to the old dogma of Morgagni we recognize now an extension to "functional" diseases where we do not (yet) find a pathological anatomical manifestation. In these cases "information" is defined as differences in physical states. They can be converted to information in the sense of meaning and purpose by communication. Pathology provides the bridge between functional genomics and morphological manifestations of diseases (phenomics). The future of pathology is based on the amalgamation of the old and reliable Morgagni-Virchow dogma with the new informational dogma. If pathology provides sense, meaning and significance to phenomics, morphology in relation to functional genomics it will retain and even enlarge its influence as the most important teaching discipline in medicine. PMID- 11894414 TI - [What does the medical student learn in pathology in Germany, China and the USA?]. AB - From the point of view of a medical student who studied at a couple of universities in Germany, one semester at a medical school in the USA, and who was involved in an exchange-program with a Chinese university, the importance of pathology as a main part of the medical course is stressed.--In Germany and the USA, pathology is a central subject in the medical course studies. After studying how the human body functions correctly (anatomy, physiology etc.), the medical student learns the pathological basics of diseases. Then clinical training, including electives or rotations, starts, where the student enters clinical settings and learns how to treat these diseases. As medicine is going to be more and more specialized, this will sooner or later also have consequences on the system of medical teaching. In the USA, for example, it is to a certain extent possible to specialize during clinical rotations already. For this reason, pathology is going to be even more important in the future.--In China there are two medical systems that exist independently: Our so-called Western Medicine and Traditional Chinese Medicine. The biggest difference in traditional medicine seems to be that diseases are treated without any (western) knowledge of their pathological background, but with the empirical knowledge of 5000 years of Chinese Medicine (so-called "black-box-system").--In conclusion, pathology is fundamental to the understanding of diseases in humans in Western Medicine. The facts taught are, of course, the same everywhere. There are differences as far as the teaching methods are concerned, both between the different countries and within Germany. Most important is the attitude of both medical students and teaching staff. PMID- 11894412 TI - [Pathology 2001: odysssey and navigation]. AB - The opening of a new millennium might be a suitable moment of looking backward and, moreover, to look forward what will happen to medicine in general and to pathology in particular during the next decades. In the 19th century, the concept of cellular pathology was ushered into medicine explaining diseases by basic cell and tissue injuries. The 20th century was characterized by a tremendous methodical progress in morphology connecting the cellular concept without a gap to present molecular medicine. In this opening address, famous pathologists are represented as examples for the different aspects of scientific and practical pathology, which has continually contributed authoritative information with clinically relevant significance to the understanding of diseases of man. The deciphering of the human genome shall have implications for several medical disciplines including pathology. However, much has to be done to find all the links between genomic abnormalities and the complete causal and formal pathogenesis of known and unknown diseases. Morphological studies carried out by the wide spectrum of methods available in modern pathology are regarded as the best effort to explain the structural aspects of the many human diseases. Taken together, the anatomic concept of disease completed by a genome-wide view shall remain the speciality of scientific medicine, so that basic and diagnostic pathology shall not perish in the future. PMID- 11894415 TI - [A new Approbation Order for teaching medical pathology]. AB - A basic reform and a draft of a new approbation order were developed during the last years to improve medical education in Germany. Aims of this reform are to link the theoretical and practical teaching, to foster the interdisciplinary teaching, to promote case- and problem-based instruction, to reduce written multiple choice examinations, and to promote oral exams. During the discussion about the reform proposals were made to put much more weight on the social and psychological aspects of health care and to reduce the teaching of a science based understanding of disease, for which pathology stands. In the final draft of the reform, however, pathology was maintained as subject of clinical education. Decisive changes now include, that the traditional distinction between general and special pathology will be abolished, and that a case- and problem-based teaching by interdisciplinary clinical-pathological conferences will be fostered. Thus the education will consist of a systematic lecture and practical sessions in pathology, which have to focus on the basic principles of the etiology, pathogenesis and classification of human diseases. Practical clinical aspects of pathology will then be thought by problem-based and interdisciplinary clinical pathological conferences or demonstrations, which start in the 4th year of the curriculum and have to be continued during the practical year's term until the end of the studies. Presently the new approbation order still requires consent of the Bundesrat. It depends on agreement about the limitation for the maximum number of students, the regulation of admittance to the medical education and the cost effects of the reform. There are some indications that solutions of these problems might be achieved during 2001. PMID- 11894416 TI - [Pathology in so-called Problem Oriented Learning (POL)]. AB - Within the evolutionary progress in biomedical research and clinical medicine the medical education plays a derminting role. Problem based learning (PBL) or problem oriented learning (POL), respectively, are main though not quite new topics in the international discussion about modern teaching in medicine. After shortly reviewing the principles of PBL the problems of this concept in general and in the special situation of the Medical Schools of German Universities bound to legal rules and with high numbers of students are discussed with special regard to Pathology representing a substantially central teaching discipline. At the Medical School of the University of Munich since 1997 the so-called Munchner Modell (Munich Model) has been successfully established representing a hybrid curriculum with integration of PBL courses and traditional forms of teaching. The experience with special regard to the participation of Pathology is discussed. IN CONCLUSION: Modern scientific medicine requires an adequate teaching and learning culture. PBL is one main concept possibly to be integrated. In the future Pathology will continuously play a central role in medical teaching because of its imminent potential. Permanent reflecting the usefulness of modern teaching proposals and critical engagement are prerequisites. PMID- 11894417 TI - [Teaching of histopathology on the Internet]. AB - The authors report their experience with the German language histopathology teaching web site www.pathologie-online.de, and discuss its design, web server statistics and future developments. The current status of German language internet resources designed for teaching of pathology at medical schools is reviewed and compared to corresponding English language resources with public access. PMID- 11894418 TI - [Pathology and clinical medicine in the 21st Century]. AB - The major task of pathology is to bridge the gap between basic science and clinical medicine. Consequently, pathology has to keep pace particularly with the progress in molecular biology and genetics and has to transfer the progress of basic research to studies of disease. Pathology has to test and continuously incorporate new methods into its scientific and diagnostic repertoire to improve diagnostic sensitivity and accuracy. The recognition of genetic alterations, particularly associated with tumors and chronic degenerative diseases, and its consequences in correlation with morphology allows the evaluation of changes in the genetic set-up and related cellular functions. This greatly improves the basis for understanding the biological background and clinical behavior of disease, thus adding a dynamic component to the snap-shot picture provided by the histologic slide. Application of genomics and proteomics will have significant impact on diagnostic pathology in the 21st century regarding definition and classification of pathologic processes, assessment of prognosis and guidance of treatment. The modern pathologist has to accept this challenge in order to maintain his position in the center of clinical medicine. However, despite all present and future advances, experience and skills in the morphologic evaluation of disease processes will still remain the "gold standard" in our field and the basis for proper application of these new technologies. When we accept the promising new techniques and tools already at the present early stage of development we may not only improve the clinical relevance of our work but gain invaluable information on the nature of diseases and the basic principles responsible for the complex morphologic pictures we enjoy in our daily work. PMID- 11894419 TI - [Cooperation between clinicians and pathologists]. AB - To the disadvantage of physician in training and quality control the number of post mortem examinations has declined in many Western countries including Germany. The frequency at which surgical or biopsy specimens are investigated by pathologist, on the other hand, appears to increase and clinicians have become increasingly more reliant on a pathologists assessment in terms of diagnosis and treatment strategy. There are some areas where communication between clinician and pathologist could be improved. The first regards the classification of malignancies which should, wherever possible, follow international guidelines such the WHO's. From a clinicians perspective assessment of tumour staging and grading should be based on the patient's prognosis rather than morphological criteria alone. In the case of rare tumours, for which the clinician can be assumed to be ignorant, reference to the literature is helpful. If the surgical or biopsy specimen reveals a malignancy that cannot be classified suggestions for a differential diagnosis can be still helpful. Frequently, at least in my field of speciality, the only tissue accessible is a hepatic or lymphatic metastasis. Determination of the tumour of origin is often essential in deciding on treatment options and every effort including immonocytochemical methods should be made to help to determine the tumour's original site. It spares enormous cost and makes many unpleasant interventional procedures redundant. Further areas where clinicians increasingly need help from pathologists are the diagnosis of inflammatory and infectious diseases and in the usage of molecular genetic methods. PMID- 11894420 TI - [Daily diagnosis by biopsy: quo vadis]. PMID- 11894421 TI - [Cytopathology--current trends]. AB - Modern imaging techniques help to detect minute tumours, even in remote locations of the body. This increases the demand to sample cellular material by minimal invasive procedures, such as fine needle aspiration (FNA). For optimal treatment, tumour cells should be characterized for prognostically relevant features. Primary and metastatic neoplasms, and individual disseminated cells of solid tumours are targets for FNA and exfoliative cytology. Recognition and typing of tumour cells is aided by immunochemistry and PCR based molecular studies. Especially immunostaining requires laboratory procedures adapted to cytology samples. Screening for precancerous lesions, highly successful in cervical cancer prevention, extends into areas such as bladder and lung cancer in selected patient groups. Cell image analysis, especially for DNA ploidy, and chromosomal FISH of interphase cells can have predictive value enhancing the information derived from the purely morphologic assessment. Prognostic tumour cell markers are studied using immunochemistry and PCR on cytological material. Cytology has to foster translational research and test the results from molecular cytogenetics, including comparative genomic hybridisation (CGH) and cDNA microarray techniques on dissociated solid tumour cells. Newly identified genetic abnormalities can be evaluated for their functional relevance and may correlate with distinct structural alterations of the tumour cells. Proper training, dedication to diagnostic cytologic work and molecular studies, and sufficient staffing will enable cytology to get the most for the patient out of limited cell material. This approach helps to distinguish genetically defined tumour subtypes by recognizing properties of the cell gestalt. PMID- 11894422 TI - [Is osteoporosis treatment cost-efficient?]. PMID- 11894423 TI - [Emergency myocardial scintigraphy after successful percutaneous coronary intervention therapy of myocardial infarction]. PMID- 11894424 TI - [Children, fat and cardiovascular diseases]. AB - Children do not normally develop atherosclerosis. However, they do develop fatty streaks in the aorta. These are reversible. During the first years of life dietary fat has an influence on blood lipids, and other traditional risk factors influence vascular function, but the consequences are unknown. As saturated fat has no positive effects, the Danish Nutrition Council recommends that the intake of saturated fat is reduced to 10 energy per cent from the age of 12 months. This can be accomplished with semi-skimmed milk (1.5% fat) instead of full-cream milk. During the first year of life, it is recommended that a teaspoon of fat is added to each serving of home made mashed food or porridge to prevent the diet from being so hypocaloric that it has a negative effect on growth. PMID- 11894425 TI - [Pharmaco-economic evaluation of drug therapy osteoporosis. A literature review]. AB - INTRODUCTION: Interventions against osteoporosis may reduce the incidence of fractures in patients and costs to society, but they also incur additional expenditure and thus call for economic evaluation. The aim of this paper was to evaluate existing literature by applying cost-effectiveness (CEA) and cost utility analyses (CUA) to pharmacological treatment of osteoporosis. MATERIAL AND METHODS: MEDLINE and the reference lists of relevant papers were searched to identify original papers on the subject. Studies were included if they were peer reviewed, written in English or a Scandinavian language, and reported CEA or CUA for a specified pharmacological intervention. RESULTS: Of the 37 identified studies, 16 met the inclusion criteria (ten CUA and six CEA), and 21 studies were excluded. Of the studies examined, 13 studies concerned hormone replacement therapy (HRT), four bisphosphonate, four calcitonin, and four calcium supplementation and/or vitamin D treatment. All were based on simulations of the long-term effects of the intervention with respect to cost and effect. However, the studies varied widely in patient selection and assumptions about duration and effectiveness of intervention, assessment of quality of life, and mortality following hip fracture. DISCUSSION: The published studies rely on limited empirical data as regards the effect of treatment, costs, and adverse effects. Several, however, indicate that some interventions may be cost-effective in high risk groups. PMID- 11894426 TI - [Advanced prehospital treatment of heart arrest by the mobile emergency unit in Aarhus. 1-year survival after out-of-hospital heart arrest--with focus on response time, survival, the given treatment and admission]. AB - INTRODUCTION: The Mobile Emergency Care Unit (MECU) in Arhus includes an experienced anaesthesiologist and a specially trained rescuer. It covers a radius of 25 km from the centre of Arhus with 330,000 inhabitants. Rescue workers in Denmark are permitted to give basic life support and defibrillation. The MECU carriers out advanced cardiac life support in accordance with "The 1998 Guidelines of the European Resuscitation Council". MATERIAL AND METHODS: Data collected by the MECU doctor on a standardised chart and survival data received from the Central Hospital Database were analysed retrospectively. RESULTS: In 1998, 4725 emergency calls were received. Twenty-five per cent of the calls were for trauma, 515 patients had cardiac disease, 158 of whom had cardiac arrest. In 86 patients, death was determined on the spot and no treatment was given. Seventy two patients received advanced cardiac life support. Twenty-five patients were admitted to hospital. Thirteen patients were alive one year later, which gives a survival rate of 52% of the patients admitted to hospital. Of the 25 patients who were resuscitated and admitted to hospital, 21 received defibrillation, 16 were intubated, 19 had adrenaline, 11 lidocaine, and 9 amidarone. Other drugs used were atropine, NaHCO3, sotalol, and CaCl. DISCUSSION: These results illustrate that for patients with out-of-hospital cardiac arrest early treatment with advanced cardiac life support performed by experienced doctors probably had a positive impact on survival, as compared to basic cardiac life support. PMID- 11894427 TI - [Prehospital treatment of patients with acute exacerbation of chronic pulmonary disease. Before and after introduction of a mobile emergency unit]. AB - INTRODUCTION: A Mobile Emergency Care Unit (MECU), manned by an anaesthesiologist and a member of the ambulance crew, was introduced in the city of Arhus in 1997. Endotracheal intubation is not performed by ambulance personnel in Denmark. The aim of this study was to describe the influence of prehospital treatment given by the MECU on the rate of endotracheal intubation, hospitalisation, and survival rate in patients suffering from acute exacerbation of chronic pulmonary disease. MATERIAL AND METHODS: We examined the data registered for patients with chronic pulmonary disease, who called for an emergency ambulance. The study covered two periods of three months: before the introduction of the MECU (September to November 1996) and after (September to November 1997). RESULTS: The study comprised 139 patients (72 patients before, 67 patients after). The MECU attended 57% of the patients. Endotracheal intubation was performed in eight patients: two before and six patients after, four of whom were intubated on the spot. Owing to the treatment given by the MECU on the spot, fewer patients were hospitalised, i.e. 50 patients (75%) versus 67 patients (93%) (p < 0.01). The survival rates were 76% before and 85% after. DISCUSSION: The MECU was a useful supplement to the ambulance service. The MECU intubated patients with acute exacerbation of pulmonary disease in the case of life-threatening respiratory failure, and in less severe cases treated the patients at home. Thus, prehospital treatment by a physician meant fewer admissions to hospital. PMID- 11894428 TI - [Resistance tests in general practice. Validity can be improved by standardized procedures]. AB - INTRODUCTION: Susceptibility testing of bacteria in urine is one of the commonest laboratory tests in general practice in Denmark. It is quick and easy to perform, but recent studies have shown low validity when the test is performed in general practice. If it is to continue as a diagnostic tool in general practice, its quality should be improved. The aim of this study was to investigate the effect of an intervention to improve the quality of susceptibility testing in general practice. MATERIAL AND METHODS: Twenty-three randomly selected general practices took part in the study. The intervention consisted of visits by laboratory technicians who instructed the practitioners in standardised procedures for susceptibility testing. Before and after the intervention, urine specimens containing monocultures of typical uropathogenic bacteria were sent to the practices. The practitioners performed susceptibility testing by the Sensicult and the Iso-Resagar methods, and the validity of the results before and after the intervention was compared. Results from susceptibility testing at the bacteriological laboratory, Odense University Hospital, were used as the gold standard. RESULTS: The median frequency of correct results increased from 82% to 98% for susceptibility testing by the Sensicult method (p = 0.001) and from 90% to 96% by the Iso-Resagar method (p = 0.05). DISCUSSION: The validity of susceptibility testing in general practice improves when preceded by instruction in standardised procedures. PMID- 11894429 TI - [Evaluation of group homes for the mentally ill in the county of Copenhagen--1980 1995. A study of a time-limited stay in the group home assessed by the development of the social situation and duration of hospitalization before and after]. AB - INTRODUCTION: We have assessed the diagnostic and social characteristics of residents, who were living in two small group homes during the period, 1 March 1980-1 August 1994. Almost all the 74 residents had a chronic psychotic disease. They had been living in the group homes for at least three months, on average 19 months. MATERIALS AND METHODS: The investigation was made from case notes and hospital records. The social characteristics were obtained from the first stay in a psychiatric ward, from two years before the patient moved into the group home to the time of moving out and 2, 5, and 10 years thereafter. RESULTS AND DISCUSSION: Almost all the patients ended up with clear symptoms of schizophrenia and were receiving a social pension. The average duration of hospitalisation for this group was reduced from one third to one seventh of the time from 2 years before to 10 years after they had left the group homes. PMID- 11894430 TI - [Standards development--I]. PMID- 11894431 TI - [Standards development--II]. PMID- 11894432 TI - [Peanut allergy]. AB - Four Danish cases of peanut allergy are described. Peanut allergy is increasing in Western countries, and is a known cause of severe reactions, some of them fatal. The prevalence in Denmark is still unknown, but it is probably under diagnosed. PMID- 11894433 TI - [How is the treatment of cerebral apoplexy to be organized?]. PMID- 11894434 TI - [Say the name!]. PMID- 11894435 TI - [Concerning the comment of Kurt Bjerregaard Stage]. PMID- 11894436 TI - [The reality is--unfortunately--"on the other side"]. PMID- 11894437 TI - [Body height among a population of children 7-18 years in the greater Copenhagen, 1981 and 1985. The Osterbro survey of children]. PMID- 11894438 TI - [Blue "marmoration" of legs as an adverse effect of vaccination]. PMID- 11894439 TI - Achalasia: a review of therapeutic options and outcomes. AB - Advances in achalasia has led to the development of new therapeutic options. This review will focus on methodology and outcomes of two established techniques; pneumatic dilation and surgical myotomy; and one new technique, LES injection of botulinum A toxin. PMID- 11894440 TI - HIV and the central nervous system. AB - Neurological complications of HIV infection are common with clinically recognized disorders ultimately affecting between 40% and 75% of patients. The spectrum of neurological disease is broad. This article highlights the common disorders of the central nervous system associated with HIV/AIDS. PMID- 11894441 TI - Homocysteine. AB - Homocystemia is a pathological condition that represents a significant role in cardiovascular morbidity and mortality. The epidemiology, pathophysiology, and clinical diagnostics and treatment aspects are described and discussed. PMID- 11894442 TI - Medical management of inflammatory bowel disease in the new millennium. AB - The medical management of inflammatory bowel disease in the new millennium requires integrating cost concerns with the efficacy and safety profiles of the expanded therapeutic options available in order to achieve optimal patient outcome. PMID- 11894443 TI - Type 2 diabetes management: a comprehensive clinical review of oral medications. AB - Diabetes is a major and growing health problem in the US. An incredible array of different oral medications is now on the market. Clinicians should understand all these new medications and in which clinical picture they will work best. PMID- 11894444 TI - Oxidant injury and antioxidant prevention: role of dietary antioxidants, minerals, and drugs in the management of coronary heart disease (Part II). AB - Antioxidants, trace minerals, and certain amino acids enhance antioxidant defense of the body by improving intracellular redox status, vascular endothelial function, and nitric oxide secretion. Antioxidant supplementation has been suggested for primary and secondary prevention of coronary heart disease. PMID- 11894445 TI - Hemoptysis: diagnosis and management. AB - Hemoptysis is a frightening and potentially life-threatening symptom. However, most cases can be approached effectively with conservative management. Bronchial artery embolization should be attempted when bleeding is refractory to medical therapy, and surgery may be needed in severe hemorrhage. PMID- 11894446 TI - Atrioventricular block revisited. AB - AV blocks, their definitions and significance, are discussed. Type II, second degree AV block is infranodal, whereas 2/3 of Type I with BBB are infranodal, 2:1 AV block is neither Type I nor II block. Infranodal blocks require pacing regardless of symptoms. PMID- 11894447 TI - Outlining the ANF's view on midwifery. PMID- 11894452 TI - Underfunding: are nurses paying the price? PMID- 11894451 TI - Boosting the image of aged care nursing. Interview by Heather Witham. PMID- 11894454 TI - Continence promotion during pregnancy. PMID- 11894455 TI - Nurses speak out about microprem babies. PMID- 11894457 TI - Informed consent and the betrayal of patient's rights. PMID- 11894458 TI - The use of syringe drivers in palliative care. PMID- 11894459 TI - Are you losing sleep over shift work? PMID- 11894460 TI - Women with hepatitis C--facts and figures. PMID- 11894463 TI - The antiseptic debate rages on. PMID- 11894466 TI - Infection control in Australia in the new millennium. PMID- 11894467 TI - Nursing observations prove critical. PMID- 11894468 TI - Achieving quality aged care. PMID- 11894469 TI - Childbirth takes its toll on mothers. PMID- 11894470 TI - Remote and rural nursing: an endangered profession? PMID- 11894471 TI - Nurses hold the key to effective pain relief. PMID- 11894472 TI - Oral glucose not so baby friendly. PMID- 11894473 TI - Alcohol misuse--perspectives from the other side. PMID- 11894474 TI - Interpreting blood glucose levels--yes it is possible! PMID- 11894477 TI - Managing withdrawal in Wodonga. PMID- 11894478 TI - Aged care: time to stop the exodus. PMID- 11894483 TI - Aspects of palliative care: pain management and hydration. PMID- 11894484 TI - Redesigning processes in ambulatory chemotherapy: creating a patient appointment scheduling system: Part 1.. PMID- 11894485 TI - Eating our young. PMID- 11894486 TI - The 2000 Schering Lecture. "Is my mom going to die?" Answering children's questions when a family member has cancer. PMID- 11894487 TI - Self-help groups: oncology nurses' perspectives. AB - During the past decade in North America, the number of self-help groups for cancer patients has grown dramatically. Nurses' knowledge and attitudes about self-help groups could influence their practice behaviours and the information they provide to cancer patients. However, little is known about oncology nurses' views regarding self-help groups. This study used a cross-sectional survey to gather information about knowledge, attitudes, and practice behaviours of Canadian oncology nurses regarding self-help groups. A total of 676 nurses completed the survey (response rate of 61.3%). The respondents had spent, on average, 21.6 years in nursing and 11.6 years in oncology nursing. Results indicated that a large majority of nurses knew about available self-help groups. Approximately one-fifth of the nurses are speaking frequently about self-help groups with patients (20.7%) and are initiating the conversation on a frequent basis (22.0%). Overall, oncology nurses rated self-help groups as helpful with regards to sharing common experiences (79.5%), sharing information (75.6%), bonding (74.0%), and feeling understood (72.0%). The most frequently identified concern regarding the groups was about misinformation being shared (37.9%), negative effects of associating with the very ill (22.1%), and promoting unconventional therapies (21.2%). Implications from the study suggest that oncology nurses would benefit from learning more about the nature of self-help groups and being able to talk with patients about the self-help experience. PMID- 11894488 TI - An evaluation of the breast self-exam (BSE) practices of Asian women. PMID- 11894490 TI - Mental health needs must be addressed. PMID- 11894489 TI - CANO Pain Education Initiative Report--Victoria Conference, October 2000. PMID- 11894494 TI - Dope 'em up and ship 'em out: issues in mental health care. PMID- 11894496 TI - Dealing with the knowledge explosion. PMID- 11894497 TI - Doing it by distance: the option for busy health professionals. PMID- 11894498 TI - Graduate course offers unique counselling experience. PMID- 11894502 TI - Informed consent. PMID- 11894504 TI - Nurses find fear in the workplace. PMID- 11894506 TI - Resuscitating aged care. PMID- 11894507 TI - Epidemiologic and outcome studies of patients who received platelet transfusions in the neonatal intensive care unit. AB - STUDY DESIGN: We conducted a historic cohort study of neonates who received platelet transfusions at the National Institute of Perinatology, Mexico City, from January 1997 to May 2000. We obtained descriptive and outcome data, and assessed demographic and laboratory means of predicting "good candidates" for a future recombinant thrombopoietin (rTpo) trial. RESULTS: A minority of the transfused patients (11.4%) received only one transfusion; the majority (88.6%) received multiple transfusions. Neonates who received one or more platelet transfusions were more likely to die (24.5% mortality) than neonates who received no platelet transfusions (3.7% mortality). Regression analyses indicated that the presence of liver disease was the best predictor of a "good candidate" for rTpo administration. CONCLUSION: The majority of neonates in our institution who receive platelet transfusions receive multiple, not single, transfusions. Receiving any platelet transfusion is a marker for high risk of death. Neonates with liver disease who receive platelet transfusions might be a reasonable group for a phase I rTpo trial. PMID- 11894508 TI - High-frequency oscillation and paralysis stabilize surfactant protein-B- deficient infants. AB - OBJECTIVE: To determine if high-frequency oscillatory ventilation and neuromuscular blockade improve oxygenation and chest radiographic appearance more effectively than high-frequency oscillation alone for surfactant protein-B (SP-B) -deficient infants. STUDY DESIGN: We reviewed medical records and chest radiographs of five SP-B--deficient infants awaiting lung transplantation. Changes in FiO2 and radiographic scores were analyzed with respect to neuromuscular blockade status. RESULTS: FiO2 consistently increased 0.20 (SD 0.11) during high-frequency ventilation without neuromuscular blockade (p = 0.02) and decreased 0.14 (SD 0.11) during high-frequency ventilation with neuromuscular blockade (p = 0.05). Chest radiographic appearance, quantified by an expansion/aeration index, consistently deteriorated without neuromuscular blockade (p = 0.01) and consistently improved with neuromuscular blockade (p = 0.03). Changes in FiO2 correlated with changes in radiograph scores (r = 0.7, p < 0.001). CONCLUSIONS: High-frequency ventilation with neuromuscular blockade optimizes oxygenation for SP-B--deficient infants. This ventilatory strategy should be considered while awaiting the diagnosis of SP-B deficiency or lung transplantation. PMID- 11894509 TI - Spiritual and religious components of patient care in the neonatal intensive care unit: sacred themes in a secular setting. AB - OBJECTIVE: We hypothesized that spiritual distress was a common, unrecognized theme for neonatal intensive care unit (NICU) care providers. STUDY DESIGN: An anonymous questionnaire form assigned to a data table in a relational database was designed. RESULTS: Surveys were completed by 66% of NICU staff. All respondents viewed a family's spiritual and religious concerns as having a place in patient care. Eighty-three percent reported praying for babies privately. Asked what theological sense they made of suffering of NICU babies, 2% replied that children do not suffer in the NICU. Regarding psychological suffering of families, the majority felt God could prevent this, with parents differing (p = 0.039) from nonparents. CONCLUSION: There exists a strong undercurrent of spirituality and religiosity in the study NICU. These data document actual religious and spiritual attitudes and practices and support a need for pastoral resources for both families and care providers. NICU care providers approach difficulties of their work potentially within a religious and spiritual rather than a uniquely secular framework. PMID- 11894510 TI - Prenatal consultation with a neonatologist prior to preterm delivery. AB - OBJECTIVE: Evaluate the usefulness of prenatal consultation with a neonatologist before preterm birth. STUDY DESIGN: A questionnaire was administered to mothers 1 week before home discharge of their preterm infant in a single regional level III neonatal intensive care unit. RESULTS: Sixty-seven mothers completed the questionnaire; 84% indicated the consult was useful and 71% were comforted by the consult. However, mothers < 30 weeks' gestation were less likely to be comforted after the consult compared to those > or = 30 weeks. Most frequently, mothers indicated that they were provided with too little detail about retinopathy of prematurity and intraventricular hemorrhage. CONCLUSIONS: Most mothers delivering a preterm infant describe the prenatal consultation with a neonatologist as useful. The majority of mothers are comforted by the information presented. PMID- 11894511 TI - Improved fluid management utilizing humidified incubators in extremely low birth weight infants. AB - OBJECTIVE: To compare fluid and electrolyte management in extremely low birth weight (ELBW) infants nursed in humidified versus nonhumidified incubators. STUDY DESIGN: Setting--tertiary intensive care nursery. Subjects--all infants with birth weight < 1000 g admitted 1/95 to 1/99 who were treated with incubators and survived for > 96 hours (N = 155). Intervention--retrospective comparison of daily weights, fluid intakes, urine outputs, and serum electrolytes between group 1 (n = 70, nonhumidified incubators, born 1/95 to 1/97) and group 2 (n = 85, humidified incubators, born 1/97 to 1/99) over the first 4 days after birth. RESULTS: Despite similar daily weight losses between groups, group 1 infants received higher fluid intakes, had lower urine outputs, and had a higher incidence of hypernatremia, hyperkalemia, and azotemia (p < 0.05). Although no differences in mortality or the incidence of patent ductus arteriosus, bronchopulmonary dysplasia, or the overall rate of nosocomial infections were observed, the proportion of gram-negative isolates increased significantly (62%, p < 0.05) following the introduction of humidified incubators. CONCLUSIONS: ELBW weight infants nursed in humidified incubators have lower fluid requirements, improved electrolyte balance, and higher urine outputs during the first 4 days after birth compared to those nursed in nonhumidified incubators. PMID- 11894512 TI - Clinical and cost-effectiveness of continuous subcutaneous terbutaline versus oral tocolytics for treatment of recurrent preterm labor in twin gestations. AB - OBJECTIVE: To compare the clinical and cost-effectiveness of treating recurrent preterm labor with continuous subcutaneous terbutaline versus oral tocolytics in twin gestations. STUDY DESIGN: In a retrospective, matched-cohort design, twin pregnancies treated as outpatients with continuous subcutaneous terbutaline were identified from a perinatal database, then matched 1:1 by gestational age at recurrent preterm labor to those receiving oral tocolytics. There were 353 patients per treatment group. A cost model was used to compare antepartum hospital, nursery, and outpatient charges. RESULTS: Infants of the subcutaneous terbutaline group had greater gestational age at delivery, higher birth weights, and less frequent neonatal intensive care unit admission. Charges for antepartum hospitalization and nursery were significantly less in the subcutaneous terbutaline group, while charges for outpatient services were less for the oral group. Mean total estimated charges were US$17,109 less for those receiving subcutaneous terbutaline. CONCLUSION: Improved clinical outcomes and decreased nursery utilization suggest cost-effectiveness of outpatient continuous subcutaneous terbutaline versus oral tocolytics for the treatment of recurrent preterm labor. PMID- 11894513 TI - When combined, early bedside head ultrasound and electroencephalography predict abnormal computerized tomography or magnetic resonance brain images obtained after extracorporeal membrane oxygenation treatment. AB - Definitive neuroimaging of the brain using computerized tomography (CT) or magnetic resonance imaging (MRI) in extracorporeal membrane oxygenation (ECMO) treated infants must be delayed until after this therapy is completed. Bedside head ultrasound (HUS) and electroencephalography (EEG) studies during ECMO, if highly correlated with later definitive neuroimaging, might be used to affect the acute clinical care and early parental counseling of infants with severe cardiorespiratory failure. One hundred and sixty ECMO-treated patients had both bedside EEG and HUS studies performed during ECMO, as well as a later CT or MRI study prior to hospital discharge. There was a significant difference in CT or MRI findings among patients having normal studies on both the HUS and EEG, compared to those having an abnormality on either the HUS or the EEG, and compared to those having abnormalities on both studies. In ECMO-treated infants, the combination of a normal bedside HUS and an EEG without marked abnormalities is highly predictive of normal post-ECMO CT and MRI neuroimaging studies. PMID- 11894515 TI - Saccharomyces cerevisiae sepsis in a 35-week-old premature infant. A case report. PMID- 11894514 TI - When betamethasone and dexamethasone are unavailable: hydrocortisone. AB - OBJECTIVE: To review the literature on alternatives to betamethasone and dexamethasone for enhancement of fetal lung maturity. STUDY DESIGN: A medline search was conducted from 1966 to the present. Trials dealing with enhancement of fetal lung maturity using modalities other than betamethasone or dexamethasone were reviewed. RESULTS: Eight studies met inclusion criteria. Excluding betamethasone and dexamethasone, the most frequently studied glucocorticoids, for fetal lung maturation, are methylprednisolone and hydrocortisone. Methylprednisolone does not cross the placenta. Two grams of hydrocortisone has been shown to improve indices of fetal lung maturity (i.e., L/S ratio) and to improve fetal outcomes compared to no treatment. CONCLUSION: There is limited information about alternatives to betamethasone and dexamethasone for the enhancement of fetal lung maturity in women at risk of preterm delivery. In the absence of these two preferred drugs, hydrocortisone can be given at a dose of 500 mg intravenously every 12 hours for four doses for this indication. PMID- 11894516 TI - Central venous catheter tip in the right atrium: a risk factor for neonatal cardiac tamponade. AB - Fatal cardiac tamponade is a well recognised complication of the use of central venous catheters in neonatal patients. There is controversy over optimum catheter tip position to balance catheter performance against risk of adverse events. We report a series of five cases of tamponade occurring in one neonatal unit over a 4-year period, related to catheter tip placement in the right atrium. Right atrial catheter angulation, curvature or looping (CA) was present in all five cases on plain radiograph. It was frequently seen in other patients over the same period. Review of the literature indicates that CA was present in 6 of the 11 previous cases where the presence or absence of CA can be determined. Where right atrial catheter tip placement is accepted, clinicians should be aware of this characteristic catheter configuration, which is a major risk factor for cardiac tamponade. We recommend that catheter tips should not be placed in the right atrium to avoid risk of tamponade. PMID- 11894517 TI - Extravasation of parenteral alimentation fluid into the renal pelvis--a complication of central venous catheter in a neonate. AB - Many complications of central venous catheters, which include perforation of the vessel walls and extravasation of the infusate into pericardial, pleural, and peritoneal cavities, have been reported. We report an infant with a central venous catheter in inferior vena cava who experienced extravasation of parenteral alimentation fluid into the right renal pelvis secondary to perforation of the renal vein. To our knowledge, this rare complication has not been reported earlier. PMID- 11894518 TI - When is meconium-stained cord actually bile-stained cord? Case report and literature review. AB - It is reasonable to assume that an umbilical cord stained green is due to the baby having passed meconium in utero. We describe a newborn baby in whom there was a delay in diagnosing an imperforate anus because the baby's umbilical cord was stained green with bile and it was assumed that the baby had passed meconium in utero. PMID- 11894520 TI - Cervical neuroblastoma, stage IV-S. PMID- 11894519 TI - Umbilical cord blood gas casebook. Interpreting umbilical cord blood gases, IX. PMID- 11894521 TI - Congenital rickets secondary to untreated maternal renal failure. PMID- 11894522 TI - Acute spontaneous hemorrhage after embolization of brain arteriovenous malformation with N-butyl cyanoacrylate. AB - PURPOSE: To investigate factors predisposing to spontaneous hemorrhage in the early period after embolization of brain arteriovenous malformations (AVMs). METHODS: The authors report 15 cases of spontaneous hemorrhage in a consecutive series of 492 brain AVMs totally or partially treated with intranidal injections of a polymerizing mixture, from 1984 to June 1998. Retrospective analysis of the records was performed with special attention to the angio-architectural features before embolization, details of embolization procedures and induced angiographic modifications. RESULTS: Pre-embolization features of greater incidence were the presence of steal phenomena (87%), multiple feeding arteries (100%), a compact aspect of the nidus (93%), and lobar topography (87%). In most cases the volume of injected glue exceeded 1 ml (80%), venous embolization was significant (67%) and immediate angiographic control showed venous stagnation in and/or around the nidus (80%). Four patients were asymptomatic (incidental discovery on systematic CT at day 3). Among the remaining 11 symptomatic patients, 6 were operated to evacuate an intracerebral hematoma, outcome was good for 4, 5 were left with sequelae and 2 died. CONCLUSION: Spontaneous hemorrhagic complications affected 3.04% of embolized patients. The combination of certain angio-architectural features, significant venous embolization and persistent venous stagnation within the nidus seem to have some predictive value of high risk hemorrhage. In light of this, additional preventive measures must be taken and a neurosurgical team systematically kept on standby. PMID- 11894523 TI - [Endovascular treatment using endoprosthesis and metallic stents for aneurysmal dissection of the intracranial vertebral artery]. AB - Dissecting aneurysms of the intracranial vertebral artery represent a more frequently recognized cause of subarachnoid hemorrhage. The poor natural history of the ruptured dissecting aneurysms indicate a surgical or endovascular treatment. Endovascular treatment usually consists of balloon occlusion of the vertebral artery. This treatment however can lead to hemodynamic complications especially when the treated artery is unique or dominant. We report two cases of ruptured dissecting aneurysms of the intracranial vertebral artery treated by intravascular stent and endosaccular GDC coils. The first patient suffered from bilateral dissection with spontaneous occlusion of the right vertebral artery and development of a pouch on the left side. The second patient suffered from a dissection of the left vertebral artery which was dominant. The stenting-coiling technique was efficient in the two cases. The patients are free of symptoms with a respective follow up of 24 and 8 months. The stenting-coiling association seems to be an interesting therapeutic option in case of intracranial dissecting aneurysms which allows a preservation of the arterial flow and selective occlusion of the aneurysmal pouch. PMID- 11894524 TI - [Exploration of the origin of supra-aortic arterial trunk: can MR angiography with gadolinium injection replace digital angiography?]. AB - Ostial stenosis of the craniocervical vessels are frequently associated with carotid stenosis. Consequently, exploration of the aortic arch is necessary prior to carotid endarterectomy. Contrast-enhanced MR angiography (gRMA) could replace digital substraction angiography (DSA). The goal of this work was to evaluate gRMA for the detection of ostial stenosis of the craniocervical vessels. Twenty two patients with carotid stenosis > 50% on sonography examination prospectively underwent gRMA and DSA. We analyzed the overall quality of each gRMA and the degree of ostial stenosis of the craniocervical vessels (innominate, left carotid, subclavian and vertebral arteries). Thirteen gRMA examination was considered as good quality and 8 as adequate for diagnosis. There was significant correlation between gRMA and DSA for degree of stenosis (k = 0.82, p < 0.0001). gRMA tends to overestimate degree of ostial stenosis, especially for vertebral arteries. We conclude that gRMA is a promising tool but cannot yet be used as a stand-alone procedure for the evaluation of ostial stenosis of the craniocervical vessels. PMID- 11894525 TI - [Study of the middle ear. Value of multiplanar reconstructions in spiral tomodensitometry]. AB - We point out the interest of computed tomographic reconstructions from spiral acquisition--particularly sagittal reconstructions--in the study of middle ear anatomy and adjacent structures: the facial canal and the chorda tympani. The reference reconstructions are axial and coronal reconstructions. So, we demonstrate the superiority of sagittal reconstructions for the visualization of the lateral process of the malleus, the body and long process of the incus, the third portion of the facial canal, and the chorda tympani. For the other structures of the middle ear and the other parts of the facial canal, these sagittal reconstructions are complementary. Besides, the best type of reconstruction to visualize the stapes and the vestibular window is the axial plane parallel to the stapes axis. PMID- 11894526 TI - [Sellar hemangiopericytoma. Report of a case]. AB - Authors report spontaneous necrosis of a sellar hemangiopericytoma, in a 44 year old man, revealed by a right cavernous syndrome. The initial CT scan showed a right sellar and parasellar spontaneous hyperdense process, intensely and homogeneously enhanced by contrast-agent. This process invaded the right cavernous sinus and extended to the sphenoid sinus. Magnetic resonance imaging showed a lesion in isointense on T1-weighted sequence and slightly hyperintense on T2-weighted sequence, with an intense and homogenous enhancement. The follow up CT scan performed 6 months later, showed a total necrosis of the lesion. Angiography showed a slightly vascularised process, with mass effect on the carotid and sylvian vessels. The sellar and parasellar localization of hemangiopericytoma is rare. The spontaneous necrosis is exceptional, but it enabled us to distinguish it from meningioma. PMID- 11894527 TI - [Primary Ewing's sarcoma of the cranial vault. Report of 2 cases]. AB - Primary Ewing sarcoma of the calvarial skull is very rare, found in less than 1% of the cases. Frontal and parietal convexities are the common sites of occurrence. We report two cases of primary Ewing sarcoma of the skull (in 13- and 14-year-old boys. The first tumor involved the right temporal region and exhibited unclear osteolytic appearance on skull x-rays while computed tomography showed an extraaxial enhanced mass and bone sclerosis with spiculated periosteal reaction. In the second case, the tumor was temporoparietal. Plain films demonstrated a large osteolysis and computed tomography revealed extensive bone destruction involving both the inner and outer tables. In both cases, the C scan revealed extraaxial enhanced masses with intracranial and extracranial extension. Radial resection was performed. Adjuvant chemotherapy was given and no recurrence or metastasis has occurred two years later. PMID- 11894528 TI - Hemangioblastoma causing cervical neural foraminal widening. AB - We present the MRI findings in a case of a 24-year-old woman with spinal hemangioblastoma, causing neural foraminal widening by producing a dumbbell mass in the lower cervical region. Hemangioblastomas can very rarely present as an intradural extramedullary lesions and this case is another exceptional pathology which should be considered among the differential diagnosis of enlarged intervertebral foramen due to neoplastic processes. PMID- 11894529 TI - [Simple cyst of the cerebellum. Report of a case]. AB - The authors report a case of simple cyst of the cerebellum documented by magnetic resonance imaging in 42-year-old men with clinical symptoms of expansive cerebellar lesion. No communication with the ventricle, no mural nodule and no enhancement after contrast injection were noted. The postoperative outcome was satisfactory after surgical removal. Histological examination of the cyst wall showed normal cerebellar tissue without epithelial lining neither tumoral features. A brief review of the seventeen cases founded in the literature is given. PMID- 11894530 TI - [Amniotic membrane graft in ocular surface disease. Prospective study with 31 cases]. AB - INTRODUCTION: Amniotic membrane's unique combination of properties including the facilitation of migration of epithelial cells, the reinforcement of basal cellular adhesion and the encouragement of epithelial differentiation [6] together with its ability to modulate stromal scarring and its anti-inflammatory and anti-bacterial activity has led to its use in the treatment of ocular surface pathology as well as an adjunct to stem cell grafts of the corneal limbus [6-4]. We report a prospective study of 30 patients so treated. MATERIAL AND METHODS: We studied 31 eyes of 30 patients subjected to amniotic membrane grafts between September 1999 and May 2000. There were 25 men and 5 women with an average age of 60.1 (range 25-86) years who were followed for a mean of 7.7 (range 4-11) months. 5 groups (A to D) were observed: A: 6 eyes. Small chronic ulcers without limbal involvement. B: 4 eyes. Ulcers of at least 75% corneal area or occupying 75% of the limbus. C: 9 eyes. Corneal burns. D: 8 eyes. Painful bullous corneal dystrophies unresponsive to other treatment. E: 4 eyes. Symblepharons. Amniotic membrane was placed on the corneal lesion, epithelial surface externally [6, 15], trimmed and sutured with interrupted 10/0 nylon, removed at one month. In two patients (11, 12) inflamed conjunctiva was recessed and amnion sutured to the recessed margin. For the bullous dystrophies we removed all the corneal epithelium and either sutured the amnion to peri-limbal conjunctiva (4 eyes) or to the limbus (4 eyes). For the symblepharons the conjunctiva was dissected to reform the fornix which was lined with amniotic membrane, sutured with 8/0 vicryl. Patients were reviewed regularity. RESULTS: Group A: All healed within 15 days, in most with dissolution of the amnion over 2-3 months although some persisted, covered with corneal epithelium. An eye with a Descemetocoele and one with a microperforation both healed. Vision improved more than two lines in 4 of 6 eyes. Group B: 2 of 4 eyes healed, one despite detachment of the membrane after 15 days. One eye was salvaged by tarsorrhaphy over a fresh keratoplasty after perforation of a neuroparalytic ulcer on failure of three successive amnion grafts. The final cornea vascularised despite an amnion graft for a meta-herpetic ulcer. Group C: 2 of 9 eyes had limbal damage in one quadrant but 7 had vessels in at least three-quarters of the circumference. One (15) also had a limbal autograft. 3 of 9 eyes healed satisfactorily with more than 2/10 improvement in acuity in each case. 2 showed further neovascularisation despite surface healing. One old chemical burn healed satisfactorily but vascularisation remained 5 eyes failed to heal with lysis of the graft, the patient who had a limbal autograft developed a vascular pannus, and in 4 eyes neovascularisation progressed to cover the entire cornea. Group D: 3 eyes settled with loss of symptoms but in 5 the graft detached within 15 days. All eyes where the membrane had been sutured to the conjunctiva beyond the limbus failed whilst 3 of 4 in which it had been sutured anterior to the limbus succeeded, leaving a persistent whitish membrane under the epithelium. Group E: We were able to reconstruct the cul de sac in 3 out of 4 eyes. In one patient with recurrent pterygium good ocular movement was restored, previously limited by scarring. One with associated ocular surface damage from a thermal burn failed by scarring of the cul de sac a month after surgery. DISCUSSION: Our best results were in persistent trophic ulcers of the cornea (Groups A and B) with a success rate of 80%, comparable to those of others [49, 37, 38]. The ready availability of amniotic membrane in our facility makes amniotic membrane transplantation the main secondary treatment for such lesions, especially because of the visual improvement we obtained. Because we did not observe any improvement in corneal thickness after this treatment we advise its early use before significant stromal lysis. The technique was not sufficient to control the effect of corneal anaesthesia in two eyes [40] or in chemical burns suggesting that amniotic membrane alone is insufficient to promote corneal healing in the absence of limbal stem cells. Nevertheless, three eyes did benefit. It has been suggested [13] that the anti-apoptotic function of amnion may prevent stem cell loss in such eyes [42], thus it appears logical to offer an amniotic membrane graft first, before stem cell transplantation, which may entrain complications in the donor eye if autografted [43] or because of the rejection risk of an allograft. It may be that an amniotic membrane graft simply becomes a holding procedure allowing time to settle the eye so as to allow secondary procedures to address the underlying cause of further damage. Our treatment of bullous dystrophy only succeeded on confining the graft to within the limbus, 3 out of 4 eyes becoming comfortable. By contrast we found amniotic membrane helpful in reconstructing symblepharons in the absence of local inflammation. CONCLUSION: Amniotic membrane grafting is a simple and straightforward surgical technique which should form part of the therapeutic arsenal for the treatment of ocular surface disease. Indications for the technique need further clarification for it is evident that it cannot correct all secondary pathology associated with limbal destruction. It is certainly preferable to conjunctival advancement and has proved useful in the reconstruction of the cul-de-sac. PMID- 11894531 TI - [Modeling the eye based on simulated refractive surgery]. AB - PURPOSE: To achieve three-dimensional modelizing of the eyeball (morphological and mechanical behavior) in order to simulate the impact of various refractive surgery techniques and to study the normal and pathological states of the eye. METHOD: Rebuilding the ocular shell is based on different kinds of imaging (MRI, ultrasound) including information provided by video topography. Image data are treated using suitable numerized filters that allow automatic segmentations of ocular globus edges. Reconstruction is based on specific mathematical functions (B-splines). The mechanical behavior of a reconstructed model is simulated by solving equations of linearized elasticity with the finitude elements method. RESULTS: Numerous simulations mimmed different refractive surgical techniques and, then validated the model. In addition, simulations of various pathologies allowed us to verify certain clinical hypotheses. CONCLUSION: This work, although still experimental, demonstrates the advantages of such simulations and will allow novice physicians an easier approach to different surgical techniques and will help them understand their effect. Furthermore, it might be useful for simulation of new surgical concepts even before their in vivo evaluation. PMID- 11894532 TI - [Pterygium: surgical treatment]. AB - PURPOSE: The surgical management of pterygium is often complicated by recurrence of disease. The goal of this study was to compare three different surgical techniques used for the treatment of pterygium. METHOD: We propose a prospective and partly retrospective study on 167 cases of pterygium 151 were primary and 16 were recurrent, treated between 1 January, 1995 and 30 June, 1998. The surgical treatment used of 3 different techniques: pterygium excision for 111 eyes, pterygium excision with conjunctival autograft for 29 eyes, pterygium excision with application of mitomycin C for 27 eyes. RESULTS: Immediate results for the three kinds of treatment were good. No serious complications were noted. Long term results show a recurrence rate of 10.3% for conjunctival autograft, 11.1% for mitomycin C application, while the pterygium excision alone had a recurrence rate of 55.9%. CONCLUSION: This study demonstrated that conjunctival autografting and the application of mitomycin C are safe surgical techniques that reduce the probability of recurrence after surgery for pterygium. They can be recommended for young people and patients exposed to sun. Mitomycin therapy is the most appropriate treatment for these cases, because of its simplicity, lower lost and the relative lack of complication. PMID- 11894533 TI - [Optical involvement in multiple sclerosis. Results of a cross-sectional study with 57 patients from Martinique]. AB - INTRODUCTION: The optical disease observed concurrent with multiple sclerosis is poorly known in blacks because of the rare occurrence of this pathology in the black population. The few studies currently available suggest more severe problems in black subjects than in Caucasians. Martinique has subjects who are genetically close or even identical but who acquired multiple sclerosis in two very different endemic areas: some in Martinique and others in France. PATIENTS AND METHODS: We performed an ophthalmological transversal descriptive study in a population of 57 black multiple sclerosis patients living in Martinique, selected according to the Poser criteria. Patients were divided into two groups: an M group (26 cases) made up of patients who had never left Martinique or the West Indies and an FM group (31 patients) made up of patients who had lived at least one year in France between the ages of 5 and 15. RESULTS: At least one episode of retrobulbar optic neuritis was observed in 16 cases (61.5%) in the M group and in 11 cases (31.5%) in the FM group. The number of eyes with either one episode or more of retrobulbar optic neuritis or papillitis was higher in the M group, 26 cases (50%) than in the FM group, 14 cases (22.6%). Multiple sclerosis began by a disease of the optic nerve in 12 cases (46%) in the M group and in 7 cases (22.6%) in the FM group. The mean visual acuity is 20/30 in the M group and 20/20 in the FM group. The mean value of the p 100 wave was 131 ms in the M group and 113 ms in the FM group. DISCUSSION: Multiple sclerosis observed in the black population living in Martinique (M group) is characterized by frequent and severe visual problems. The visual phenotype of the FM group is quite similar to the visual phenotype of Caucasians. More than ethnic and genetic factors, the tropical area of acquisition should have an influence on the visual phenotype. For unknown reasons, the optical problems appear severe when multiple sclerosis is acquired in a low endemic area. PMID- 11894535 TI - [Value of MRI in the treatment of Grave's disease orbital myopathy]. AB - In order to evaluate the predictability of the results in the treatment of myopathy in cases with the clinical signs of muscle involvement, 177 extraocular muscles of 27 cases whose oedematous status was detected by MRI and who were given antiinflammatory treatment according to the data of this method, were studied. The nature of involvement was detected in respect with the signal intensity and thickness of each rectus muscle prior to the treatment and at the end of the sixth month following a three months' application of combined treatment of steroids and irradiation of 2000 rads. When the initial and final results were compared, the signal intensities of four involved recti showed significant decrease at the end of the treatment, as they were evaluated separately or together. Besides the thicknesses of these groups of involved recti which were evaluated separately showed significant decrease. The evaluation of the signal intensities by MRI is a way that enables noninvasive detection of the edema and prediction of the anti-inflammatory treatment's results of dysthyroid myopathy. Therefore a systematic follow up by MRI is recommended for the treatment choice in dysthyroid myopathy. PMID- 11894534 TI - [Enucleation with on-the-table evisceration. Is it a good technique?]. AB - PURPOSE: To evaluate the surgical technique of enucleation followed by an "on-the table evisceration" and placement of a hydroxyapatite orbital implant wrapped by the patient's own sclera for the treatment of blind phthisis painful eyes. PATIENTS AND METHODS: In this single-center retrospective study, 50 consecutive patients undergoing an operation using the same surgical technique, between April 1993 and November 1999, were studied. Patients underwent enucleation, then the eyeball was eviscerated "on the table". The patient's own cleaned sclera was used to wrap a hydroxyapatite orbital implant, the posterior pole of the sclera was placed at the anterior pole of the implant. Conjunctival breakdown, sphere size, conjunctival discharge, the first signs of sympathetic ophthalmia motility, and cosmetic results were analysed. RESULTS: After an average follow-up of 13.3 months few complications were encountered: 4 cases (8%) of inclusion cyst and 3 cases (6%) of discharge. The implant placed had a diameter of 18 mm, 20 mm, 22 mm in, respectively, 48%, 48%, and 4% of the eyes. The prosthesis motility was good, medium, and poor in, respectively, 33 (78.6%) cases, 8 (19%) cases, and 1 (2.4%) case. The prosthesis tolerance was good, medium, and poor in respectively 86%, 10%, and 4% of the cases. CONCLUSION: The surgical technique of enucleation followed by an "on-the-table" evisceration and autologous sclera wrapping a hydroxyapatite implant is an easy procedure. It allows, on phthisis eyeballs, the placement of a large orbital implant for good cosmesis results, without major complications. PMID- 11894536 TI - [Hereditary hyperferritinemia syndrome and cataract]. AB - Hereditary hyperferritinemia cataract is a recently described autosomal dominant syndrome, characterized by bilateral cataracts and elevated level of serum ferritin. PATIENTS: Three members of a family were investigated for cataract and hyperferritinemia. A 30-year-old woman had elevated serum ferritin levels and bilateral cataracts. She was treated for hemochromatosis, but serum iron and transferrin saturation were normal. Her two sons, nine and five years old, also had a high ferritin level and bilateral cataracts. RESULTS: The ferritin level was 1200 micrograms/L in the woman's serum, and respectively, 974 and 965 micrograms/L in the two boys' serum. The mother had a visual acuity of 8/10 in the right eye and 5/10 in the left eye. The cataract comprised fine crystalline cortical opacities, extending axially. The two sons had 7 to 8/10 in both eyes. No other ophthalmic abnormality was noted. These patients were heterozygous for a 16 bp deletion on the L-ferritin gene. DISCUSSION: Ferritin is an iron storage ubiquitous protein present in every cell. In hyperferritinemia cataract syndrome, serum iron and transferrin saturation are normal, and the elevated serum ferritin level is the consequence of an autosomal dominant disorder. The cataract is made up of the accumulation of small opacities disposed radially and more numerous on the outside edges, with relatively good visual acuity. The size of the cataract seems to be correlated to the serum ferritin level. In hemochromatosis, hyperferritinemia is related to increased iron stores and is not associated with cataracts. PMID- 11894537 TI - [Total anterior opacification on the anterior surface of hydrophilic acrylic implanted lens]. AB - We report a case of cloudiness occurring on the anterior surface of a hydrophilic acrylic foldable intraocular lens 1 year following implantation. This was not a case of phimosis of the capsulorhexis; the dondiness seemed secondary to fibroepithelial proliferation. Successful reopening was obtained with YAG laser. No recurrence was observed during the six months of follow-up. PMID- 11894538 TI - [Optical coherence tomography in the follow up of macular edema treatment in retinitis pigmentosa]. AB - Macular edema is frequently responsible for loss of central vision in patients affected with retinitis pigmentosa (RP). This macular edema can be treated with acetazolamide. Our purpose was to evaluate optical coherence tomography (OCT) examination in the follow-up of macular edema in RP. In addition, we tried to evaluate the minimal efficient dose of acetazolamide, using means other than fundus examination or fluorescein angiography. We report the cases of 5 patients affected with typical retinitis pigmentosa and fundus appearance of macular edema. These patients received oral acetazolamide treatment (500 mg/d). The OCT examinations were performed before and during treatment, which allowed us to demonstrate, quantify and monitor the progression of macular edema during treatment. OCT appears to be a useful tool in the follow-up of patients affected with macular edema and RP. This noninvasive examination contributes to improving our strategy in treating patients. PMID- 11894539 TI - [Acute ischemic optical neuropathy which became bilateral in 3 days in Horton disease]. AB - We report a case of a 71-year-old man with sequential visual loss within 3 days due to giant cell arteritis confirmed by a temporal artery biopsy. The anterior ischemic optic neuropathy was associated with cilioretinal artery occlusion on the right eye. He improved after intravenous corticosteroid therapy. Clinical characteristics and treatment are discussed. PMID- 11894540 TI - [Orbital exenteration]. AB - Exenteration of the orbit is a disfiguring and destructive procedure that is usually reserved for treatment of life-threatening orbital malignancy when a less radical operation such as local surgery, chemotherapy, or irradiation are deemed inadequate or have failed. Many methods have been published for managing the socket, but of primary importance is the need to remove all diseased tissue prior to considering any reconstructive efforts. Options include spontaneous granulation, skin grafting, or muscle flaps. This article will describe the development and the indications for this procedure and will outline the surgical techniques and its complications, the various reconstructive efforts, and survival. The success of orbital exenteration depends on recurrence, histological type, tumor size, and tumor-free margins. PMID- 11894541 TI - [Age-related macular degeneration and genetics]. AB - Age related macular degeneration (AMD) is the main cause of blindness after age 55 in western countries. These last past years, several lines of evidence such as familial aggregation or twin studies suggested a genetic component in AMD. However, the late onset of the disease and the fact that AMD is a polygenic and multifactorial disease are the main limiting factors for linkage studies. Gene candidate strategy allowed the exclusion of several genes (VMD2, RDS, TIMP3) and lead to the implication of two genetic factors: the apoE gene (involved in the transport of lipids) and the ABCR gene (involved in Stargardt macular dystrophy). Concerning the apoE gene, a lower frequency of the epsilon 4 alleles carriers was observed in the exudative AMD population compared with controls, supporting the idea of a role of the apoE gene in exudative AMD with drusen. These results, together with ultrastructural studies, suggest that allele epsilon 4 is a protective factor for drusen and thus for AMD. For the ABCR gene, several studies and a large multicentric study definitively show that some heterozygous mutations are predisposing factors for AMD, in a polygenic and multifactorial model. PMID- 11894542 TI - [Evisceration using the Russian doll or the parachute technique]. AB - The authors describe two new techniques of evisceration after resection of the corneal epithelium and limbus with a conservative approach of the posterior lamella of the cornea and anterior sclera after a 360 degrees dissection of the sclera behind the insertion of the oculomotor recti muscles, and with a conservative approach of the recti oculomotor muscles. The technique is called "Parachute" when the posterior sclera is removed and "Russian Doll" when the posterior sclera is not removed and placed behind the orbital implant. The implant is then introduced with "bird cage" forceps. The indications of the technique are those of evisceration when the size of the remaining globe is at least one-third of the volume of a normal eye. The main advantage is to provide safely a large volume for the implant in order to prevent post enucleation socket syndrome. PMID- 11894543 TI - [Limbal dermoid and Goldenhar syndrome. Report of an anatomoclinical study]. AB - Dysembryoplastic neoplasm, the limbal dermoid is a benign congenital tumor involving the outer coat of the eye. This congenital tumor affected the right eye of a 6-year-old girl. The lesion was typically hemispheric, covered with pink skin, located in the inferior and temporal part of the cornea, and devoid of any major consequences except the cosmetic appearance of the eye. There were no other major abnormalities in the clinical examination but a single skin tag could be observed at the homolateral preauricular area without any visible cutaneous fistula next to it. This additional change allowed us to establish the diagnosis of a minor form of Goldenhar syndrome. This malformative syndrome typically includes several changes: dermoid and/or dermolipoma, preauricular tags and/or cutaneous fistulas in the same area, vertebral abnormalities etc. These findings may be limited to dermoid and cutaneous tags, as in our report. Such changes must be looked for in all limbal dermoids. PMID- 11894544 TI - [Conjunctivo-corneal complex choristoma. Report of an anatomoclinical study]. AB - Complex choristomas are very rare ocular tumors. We report a case of a 3-month old infant who was referred to us because of a left congenital conjunctivo corneal protruding mass. Total excision removing 2/3 of the limbus was performed and replaced simultaneously by a limbal autograft. Histopathological examination of the excised choristoma revealed many pilosebaceous follicules in the conjunctival tissue, keratinized epithelium, lacrimal glands, and differentiated cartilage. PMID- 11894545 TI - [Value of sonography in the diagnosis and follow-up of patients with cryptogenic inflammatory bowel diseases in current practice: review of a 10-year experience in a community hospital]. AB - Inflammatory bowel diseases (IBD) are chronic inflammatory diseases of the bowel that are of unknown etiology. These diseases either progress with intermittent flare-ups interrupted by periods of remission or on a chronic active progressive mode. IBDs include Crohn disease (CD) and ulcerative colitis (UC). Clinical and imaging diagnosis often is challenging, hence explaining the frequent time delay between onset of disease and initiation of therapy. Clinical evaluation is characterized by three consecutive steps: consider a diagnosis of IBD; exclude other causes of inflammatory bowel disease; differentiate CD from UC since a definitive curative surgical treatment is available for UC. US is non-invasive, widely available, easy to perform, and relatively inexpensive and thus represents a significant advance in the evaluation of these three steps. The role of US in the evaluation of patients with suspected IBD will be reviewed. PMID- 11894546 TI - [Color Doppler sonography of superficial capillary hemangiomas]. AB - PURPOSE: Retrospective analysis of the morphologic and hemodynamic characteristics of hemangiomas in infants provided by color Doppler sonography. MATERIALS AND METHODS: 94 hemangiomas were studied in 87 children separated into four classes of age (0-3 months, 3-6 months, 6-12 months, > 12 months). Eleven were followed with several examinations. Echogenicity, compared to normal subcutaneous tissues, and degree of vascularization were assessed qualitatively. Resistive index (RI) was measured in 78 cases. RESULTS: All lesions presented as vascularized solid masses: 63% were hypoechoic, 16% hyperechoic and 21% mixed. All hypoechoic and mixed lesions showed higher vascularity than hyperechoic ones. Hyperechoic hemanigomas were more frequent in the fourth class of age. Mean resistive index was significantly higher in the first (0.61 +/- 0.14) and the fourth (0.6 +/- 0.14) classes of age than in the second (0.51 +/- 0.12) (p = 0.01 and < 0.03, respectively), and higher in the hyperechoic group (0.7 +/- 0.13) than in the hypoechoic group (0.53 +/- 0.11), (p < 0.01). Seven of 11 cases moved from hypoechoic to hyperechoic or mixed. Increase of resistive index with age was noted in 9/11 cases. CONCLUSIONS: Superficial hemangiomas have typical but variable gray-scale, spectral and color flow sonographic patterns. These fluctuations are probably related to phases of evolution. PMID- 11894547 TI - [Quantification of hepatic arterial and portal venous flow using ultrasound contrast agents for early detection of liver metastases of colorectal cancers]. AB - PURPOSE: To quantify liver blood flow using US contrast agents and to evaluate arterial and portal changes in control patients and patients with liver metastases. MATERIALS AND METHODS: Twenty eight patients were included in this study, 8 controls (M0) and 20 patients with liver metastases from colon carcinoma (M+). Hepatic blood flow from hepatic artery and portal vein were determined using quantification of enhancement after contrast injection using Power Doppler US. The ratio of enhancement rise from artery and vein allows calculation of Contrast Enhanced Doppler Perfusion Index (CEDPI) as previously described for Doppler Perfusion Index (DPI). RESULTS: A significant difference was noted for CEDPI between controls (0.49 +/- 0.07) and liver with metastases (0.70 +/- 0.12). CONCLUSION: This functional method of evaluation of liver blood flow was easy to perform, and would be valuable for early detection of overt micro-metastases before anatomical changes observed by conventional imaging. This is helpful for accurate staging of colon carcinoma. PMID- 11894548 TI - [Doppler ultrasonography evaluation of paraumbilical vein and portal hypertension]. AB - Portal hypertension is associated to the development of portosystemic collateral veins, particularly the paraumbilical vein. PURPOSE: To evaluate the biometric and hemodynamic characteristics of the portal vessels related to the presence of a patent paraumbilical vein, in the setting of portal hypertension secondary to hepatosplenic schistosomiasis. METHODS: 75 patients with portal hypertension secondary to hepatosplenic schistosomiasis were evaluated by Doppler US. The patients were studied based on the presence (group B) or not (Group A) of a patent paraumbilical vein. The diameter and blood flow velocity of the portal vessels and of the paraumbilical vein were recorded. RESULTS: The paraumbilical vein was detected in 17.33% of patients. The results showed an increase of the diameter of the main and left portal vessels whenever a patent paraumbilical vein was present (portal vein: A = 1.14 +/- 0.29 cm/B = 1.33 +/- 0.16 cm; left branch: A = 0.95 +/- 0.25 cm/B = 1.30 +/- 0.24 cm). The mean blood flow velocity was also increased in the portal trunk (A = 15.96 +/- 6.17 cm/sec/B = 19.82 +/- 6.26 cm/sec) and in the left portal branch (A = 14.77 +/- 4.29 cm/sec/B = 19.92 +/- 6.88 cm/sec). CONCLUSION: The presence of a patent paraumbilical vein is related to significant biometric and hemodynamic variations in the portal venous system, in the setting of portal hypertension secondary to hepatosplenic schistosomiasis. PMID- 11894549 TI - [132 grams of tamoxifen: ultrasonographic and MRI appearance of endometrial carcinoma]. AB - Endometrial carcinoma is a rare iatrogenic complication due to the adverse estrogenic like effect of Tamoxifen on the uterine mucosa. We report the delayed case of an endometrial carcinoma after an unusual twleve year long daily administration of Tamoxifen (cumulative dose = 131 g). Endovaginal contrast ultrasound examination (Levovist, Schering, Germany) and MRI appearances are described. PMID- 11894550 TI - [Splenic nodules and sickle cell anemia]. AB - OBJECTIVE: We report 4 patients with sickle cell anemia presenting with intra splenic benign nodules corresponding to islands of preserved tissue within splenic ferro-calcinosis. MATERIAL AND METHODS: Ultrasound, CT and MRI findings were evaluated and compared to a follow-up study by ultrasound and CT done after 6 to 12 months. RESULTS: Ultrasound showed multiple well-defined rounded nodules appearing hypoechoic compared to the rest of the spleen that was hyperechoic. On CT, the nodules were homogenous, hypodense relative to the spleen, isodense to the liver in 3 cases and hypodense to the liver in 1 case. On MRI, the nodules appeared relatively hyperintense within low-signal-intensity spleens. The ultrasound and CT follow-up study demonstrated no remarkable change. CONCLUSION: In sickle cell patients, intra-splenic benign nodules corresponding to normal splenic tissue may be identified on imaging studies. The differential diagnosis is discussed. PMID- 11894551 TI - [Diagnosis and follow-up of a large intrahepatic portocaval fistula in a newborn]. AB - Portocaval fistulas are rare and only exceptionally discovered in newborns. We report the case of a large portocaval fistula associated with portal hypoperfusion detected at Doppler US imaging in an otherwise asymptomatic 5 week old infant. The patient remained asymptomatic over the following two years. At that time, preoperative angiogram showed a normal portal venous system and the fistula was surgically closed. Postoperative US showed a normal and patent portal system, without evidence of portal hypertension. PMID- 11894552 TI - [Chronic arterial mesenteric ischemia: Doppler color ultrasonography demonstration of collateral flow]. AB - Two cases of atypical mesenteric ischemia where color Doppler US demonstration of the underlying arterial abnormality and collateral supply was possible are presented. Significant stenosis of the celiac axis and thrombosis of the SMA were clearly depicted, along with the presence of collateral arterial supply. Endovascular treatment was successful in both cases. These cases confirm the possibility of detecting collateral flow at Doppler imaging in patients with mesenteric ischemia, both for diagnosis of mesenteric ischemia and endovascular treatment planning. PMID- 11894553 TI - [Warm saline enema for reduction of intestinal invagination under the ultrasonographic guidance: preliminary results apropos of 2 cases]. AB - The authors describe the use of isotonic normal saline enema under US guidance in the treatment of acute uncomplicated intestinal intussusception in infants. The technique was successfully used in two patients aged respectively 18 and 16 months, with 11 and 9-month follow-up. The various steps of this technique and the results are analyzed and discussed. It is a valuable alternative to surgical treatment or hydrostatic reduction with opaque x-ray contrast under fluoroscopic guidance. PMID- 11894555 TI - To Russia with love. PMID- 11894556 TI - Defending the public health system. PMID- 11894559 TI - Nursing information and the use of electronic health records. PMID- 11894558 TI - Emergency contraception still an unknown quantity. PMID- 11894561 TI - Stem cell transplantation in children. PMID- 11894563 TI - Turning points in addiction. Interview by Fiona Armstrong. PMID- 11894562 TI - RN education has found its level. PMID- 11894564 TI - Facing reality: the transition from student to graduate nurse. PMID- 11894567 TI - Nurses retain core position in the multidisciplinary team. PMID- 11894568 TI - Creating Eden for aged care. PMID- 11894569 TI - Nursing in Taiwan. AB - Although 10 years ago there was a crucial nursing shortage, Taiwan is currently experiencing an oversupply due to recent education reforms. Many new junior colleges have now been approved to take nursing courses. Taiwanese nurses are graduating at greater numbers than are leaving the profession, causing an ever increasing surplus. PMID- 11894566 TI - Residents with dementia respond to pampering. PMID- 11894570 TI - Menopause: choices for women. PMID- 11894572 TI - Australia's nurses breaking new ground. PMID- 11894573 TI - A place for complementary health care? PMID- 11894574 TI - Asthma and the Buteyko breathing method. PMID- 11894576 TI - Nursing and naturopathy: the challenge of multiparadigm education. PMID- 11894581 TI - Writing skills: capturing the elusive elements. PMID- 11894582 TI - The utilization and efficiency of the informal caregivers' coping strategies. AB - This study highlights the coping strategies used by informal caregivers whose husbands live with cancer. It also aims at measuring the efficiency of the selected strategies. The convenience sample was composed of 30 informal caregivers. The results indicate that informal caregivers primarily use support, optimism, independence, and facing of the situation. In general, the categories of coping strategies most often used by informal caregivers are considered by them to be the most efficient. The results of the study encourage nurses to identify more regularly the coping strategies used by informal caregivers; to recognize their efficiency and implement interventions likely to improve the informal caregivers' stress management. PMID- 11894579 TI - Do rituals still rule over research evidence? PMID- 11894583 TI - Linking nursing pain assessment, decision-making and documentation. AB - A clinical nurse specialist's (CNS) experience in the development and implementation of a pain assessment and treatment flowsheet (PATF) to enhance the nursing assessment, decision-making, and documentation of pain on a palliative care unit in a community hospital is described in this article. Members of the palliative care interdisciplinary team use the PATF for clinical decision-making in the day-to-day management of patients' pain. The PATF is undergoing revision and re-implementation to promote the utilization of the tool beyond the specialty of palliative care and into the general patient population. PMID- 11894586 TI - Bringing oncology research into the clinical setting: meeting the standards. PMID- 11894585 TI - The Ontario Breast Screening Program -- making a difference through screening and early detection. . PMID- 11894587 TI - The 2000 Helene Hudson Memorial Lecture. Decisional role in seriously ill hospitalized patients near the end of life: the patient's and provider's perspective. AB - Decisions about whether or not to implement life-sustaining therapies are complex and are becoming more so as the ability to prolong life with advanced technologies and care increases. The objectives of this study were: (1) to determine seriously ill hospitalized patients' preferences for decisional role with respect to decisions about life-sustaining treatments, and (2) to determine if providers were aware of patients' preferences. This prospective, descriptive pilot study was conducted at an Ontario teaching hospital. One hundred and seventeen seriously ill adult patients admitted with cancer and non-cancerous conditions participated in a structured interview. Fifty-three nurses and 63 physicians responsible for the care of the participating patients also participated. Patients and providers were asked similar questions about end-of life discussions and preference for decisional responsibility for life-sustaining treatments. Most patients (n = 89, 77%) had thought about end-of-life issues and were willing to discuss these with their physicians and nurses, but few (n = 37, 37%) reported such discussions. Preferences for decisional role varied; most indicated a preference for a shared role (n = 80, 80%) and there were no differences in patients with or without cancer. Generally, both physicians and nurses were not aware of or did not determine accurately patient preferences for decisional role. The findings from this study show that seriously ill hospitalized patients have thought about and are willing to share in discussions about end-of-life care with their providers, yet many have not. PMID- 11894588 TI - The schizophrenic mind. PMID- 11894590 TI - [Eating disorders--a challenge for the next level?]. PMID- 11894589 TI - The man behind a beautiful mind. PMID- 11894591 TI - [Endovascular treatment of aortic aneurysms]. PMID- 11894592 TI - [Treatment of aortic aneurysm--new or old method?]. PMID- 11894593 TI - [Treatment of bulimia nervosa--results from Modum Bads Nervesanatorium]. AB - BACKGROUND: Hospital admission for bulimia nervosa is rather uncommon, but may be indicated in cases of psychiatric comorbidity, long duration of treatment and previous treatment failures. We describe a multicomponent inpatient treatment programme consisting of cognitive-behavioural group and individual therapy, physical training and steps to normalize eating patterns. MATERIALS AND METHODS: All 47 patients treated between 1998 and 2000 were studied. Patients were interviewed and completed self-report instruments at admission and discharge. RESULTS: At discharge, a significant improvement with respect to bulimic as well as general psychiatric symptoms had occurred. INTERPRETATIONS: The results may indicate that improvement occurs even for severe bulimia with personality disorders, and that hospital treatment may be needed to accomplish this kind of change. Follow-up studies are necessary in order to corroborate these findings. PMID- 11894594 TI - [Cerebral computer tomography in subarachnoid hemorrhage]. AB - BACKGROUND: A cerebral CT scan is routinely performed in all patients evaluated for subarachnoidal haemorrhage. Quick and accurate diagnosis is of the utmost importance in such patients, but the accuracy and value of the initial CT scan has not been fully established. MATERIAL AND METHODS: Initial CT scans of 70 patients (45 women) with subarachnoid haemorrhage were reviewed retrospectively. Cerebral CT scans were performed without intravenous contrast and evaluated independently by two observers. RESULTS: CT scans were positive for blood in the cerebrospinal fluid spaces in 64 out of 70 patients (91%). Blood was most frequently seen in basal cisterns (75%), Sylvian Fissure (73%) and cerebral cortical sulci (67%). Evidence of raised intracranial pressure was present in 26 patients (41%). The site of the ruptured aneurysm could be localised by CT with high accuracy only in ruptured aneurysms of the middle cerebral artery. The median time from symptom onset to examination was significantly higher in patients with a normal than a pathological CT scan (87 hours vs. 4 hours, p < 0.001). A lumbar puncture was positive for blood in all six patients with a normal CT scan. INTERPRETATION: This study demonstrates that a lumbal puncture should be performed after a normal cerebral CT scan if subarachnoid haemorrhage is clinically suspected. PMID- 11894596 TI - [Phenytoin poisoning caused by interaction with ticlopidine]. AB - BACKGROUND: Drug interactions may cause diverse clinical symptoms. MATERIAL AND METHODS: A case history. RESULTS: A 58-year-old man was admitted to hospital with suspected stroke. It turned out that his neurological symptoms were caused by phenytoin intoxication. The patient had previously taken phenytoin for epilepsy for nine years without being intoxicated. Hospital records revealed that he had received ticlopidine for two weeks in connection with a coronary stent procedure five to seven weeks earlier. The probable cause of the phenytoin intoxication was interaction between ticlopidine and phenytoin. INTERPRETATION: This case history demonstrates that apparently unrelated drugs may interact and result in serious toxicity. PMID- 11894595 TI - [Endovascular stent-graft repair of abdominal aortic aneurysm]. AB - BACKGROUND: Endovascular stent-graft repair of abdominal aortic aneurysms was introduced in 1991. Long-term results from randomised studies are still not available. Aneurysm ruptures after stent-graft repair have been reported, and there has been a considerable need for redo procedures. MATERIAL AND RESULTS: At Rikshospitalet in Oslo, Norway, 26 stent-graft implantations for abdominal aortic aneurysms were performed during the years 1996-2000, two of which were primary technical failures immediately converted to open repair. During follow-up, two patients have died from unrelated causes after six and 12 months. Eight patients were converted to open repair 8-50 months (median 31) after implantation. Indications for conversion were migration (n = 4), increasing aneurysm diameter (n = 3) and rupture (n = 1). Of the 14 patients still under observation, six have had one or more endovascular interventions (a total of 10) for failing graft. INTERPRETATION: Continuous technology development has been used as an argument to postpone randomised studies, as improved results are expected with new generations of stent-grafts. There is a need for discussion of the strategy for the further use of endovascular repair of abdominal aortic aneurysm. PMID- 11894597 TI - [New guidelines for basic and advanced resuscitation of adults and children]. AB - We present the latest changes in the guidelines for resuscitation from the International Liaison Committee on Resuscitation (ILCOR). Defibrillation performed by non-medical personnel is more strongly endorsed than before. In unintubated patients the ventilation-to-compression ratio should always be 2:15. With a foreign body in the airway of an unconscious patient, compressions should be performed instead of the Heimlich manoeuvre. Amiodarone 300 mg intravenously is the first choice in refractory ventricular fibrillation or ventricular tachycardia in adults. Intubation of children is only recommended when performed by experienced personnel. The use of drugs is less recommended than previously; albumin is not recommended for the newly born. PMID- 11894598 TI - [Eating disorders--how should treatment be organized?]. AB - BACKGROUND: Well controlled normal population studies show no sharp increase in the incidence and prevalence of eating disorders. However, more individuals seem to seek treatment, and there is a need for more precise estimates of the expected patient load at various levels of care. MATERIALS AND METHODS: The total number of potential patients was estimated on the basis of data on the number of women aged 15-44 years, the number of hospitals and outpatient clinics per county in Norway, working hours per year for general practitioners, and the prevalence of eating disorders. RESULTS: In total, about 50,000 Norwegian women may suffer from eating disorders; about 600 may need highly specialized services. At the most, each outpatient clinic may expect about 80 annual referrals for bulimia nervosa and binge eating disorder, and each hospital may expect 10-20 patients with bulimia nervosa. On average, each outpatient clinic and hospital may expect 8-9 referrals of patients with anorexia nervosa. INTERPRETATION: There is a need for increased treatment capacity, better clinical skills and better organization of treatment services for patients with eating disorders. PMID- 11894599 TI - [Vascular surgical patients and patient rights legislation]. AB - BACKGROUND: From 1 January 2001, a new Patients Rights' Act came into force in Norway that regulates many aspects of clinical work, including patients' right to be adequately informed, the right to a second opinion, the right to choose hospital, the right to refuse a blood transfusion, and the terminally ill patient's right to refuse further treatment. This study aims at assessing the impact of the new legislation on given cases in vascular surgery and to clarify in what ways work procedures in a vascular surgical department should be changed in view of the new rules. MATERIAL AND METHODS: Ten cases drawn from our department between autumn 2000 and January 2001 were analysed in relation to the new legislation, each case representing a specific medico-legal issue regulated by the new act. RESULT: By and large, established practice in our department is in line with the new legislation. However, better documentation is required, as well as more emphasis on integrity in relation to competing hospitals. INTERPRETATION: Surgeons should be well informed on regulations directly affecting the practice of surgery. PMID- 11894600 TI - [The dying patient's preferences when end-of-life decisions are made]. AB - BACKGROUND, MATERIAL AND METHODS: In this presentation we discuss end-of-life decisions on the basis of the history of a young patient with motor neuron disease and pneumonia. RESULTS: The patient wanted treatment for an acute pneumonia. Weaning from the respirator turned out to be difficult and the patient's condition was complicated by pneumothorax. He refused further treatment and managed to disconnect the respirator. He even tried self-extubation, but did not succeed, as his muscle strength was impaired. After two days the pneumothorax had spontaneously resorbed and the patient was awake. He refused further treatment and demanded that all therapy be stopped. A meeting was held of the ad hoc established local clinical ethical committee. The patient was conscious, mentally alert and well informed. The respirator was stopped and the orotracheal tube removed. The physician and nurse stayed bedside, ready to intervene if the patient wanted the ventilator treatment re-established, or to give pharmacological treatment for pain, respiratory distress, or anxiety. The patient was calm for five minutes, after that he started to feel respiratory distress. After a short while consciousness was lost, and he died after 30 minutes. INTERPRETATION: Ethical challenges are discussed. The story illustrates the importance of good communication between patient and doctor to facilitate the patient's autonomy. PMID- 11894601 TI - [Nic Waal--the mother of Norwegian child and adolescent psychiatry]. AB - Nic (Caroline) Waal (1905-60) is said to be the mother of Norwegian child and adolescent psychiatry. She was an engaged and engaging personality, outgoing, at times aggressive and at the same time very sensitive. Her sensitivity caused her suffering but it was an asset in her work with patients. She qualified in medicine in Oslo in 1930 and received her psychoanalytic training in Berlin and Oslo. She was certified as a psychoanalyst in 1933, as a psychiatrist in 1951 and -among the first in Norway--as a child psychiatrist in 1953. From 1953 till her untimely death in 1960 she was head of Nic Waal's Institute, a treatment centre and training institute for all child psychiatric professions. Through the training she provided and her published work she has had considerable influence on the development of child psychiatry in Norway and in Scandinavia. She also developed her own method of psychodiagnosis through the study of patients' pattern of muscular tensions. Her institute is still active and expanding, having included family therapy and neuropsychology and neuropsychiatry. PMID- 11894602 TI - [Diagnosis of drug-induced allergic reactions]. PMID- 11894603 TI - [Nicotine dependence--medico-biological aspects]. AB - The nicotine in tobacco products is strongly addictive. This was generally recognised no earlier than in the late 1970s, though it was well known within the international tobacco industry in the early 1960s. Nicotine acts as an addictive substance by binding to acetylcholine receptors and causing the release of dopamine in the brain, though other signalling substances are also important for the action of nicotine in the central nervous system. Withdrawal syndrome is the typical evidence of physical addiction to nicotine. Nicotine addiction can develop rapidly. There are, however, individual differences; genetic predisposition may have a bearing on these differences. PMID- 11894604 TI - [Smoking as a theme in popular health literature 1940-1960]. AB - BACKGROUND: Little is known about the information available to smokers who started smoking in the early days of research on smoking and health. The main topics of investigation in our study were: How much information on the health hazards of smoking was available to non-professionals in Norway between 1940 and 1960? What characterised this information, and did it change in quantity or quality over this period? MATERIALS AND METHODS: Representative samples of health magazines and books from 1940 to 1960 were analysed for information on the health effects of smoking. Frequency and size of articles were registered and a basic statistical analysis of the information performed. RESULTS: There was a slight increase in the amount of literature published on the subject. Religious publications were the most direct in their communication and the most consistent in recommending non-smoking. Compared to other publications, they eagerly speculated on underlying physiological mechanisms. The non-religious publications seem to better reflect the general discussions and disagreements in the field; according to several, only "exaggerated" smoking constituted a threat to health; "moderate consumption" was considered harmless. INTERPRETATION: While religious literature engaged in overt and sometimes undue speculation, the non-religious publications in the sample seemed to reflect a belief in the harmlessness of a "normal" consumption of cigarettes. The debates and disagreements in the scientific community are also more apparent in the non-religious publications. PMID- 11894605 TI - [What messages did the Norwegian tobacco industry communicate to consumers in its advertising?]. AB - BACKGROUND: A Norwegian official report, NOU 2000: 16, Tort liability for the Norwegian tobacco industry, concludes that Norwegian law and judicial practice allows lawsuits against the tobacco industry for damages. A crucial claim in such suits would be that the industry withheld information on health risks and addiction and instead used advertising messages that undermined the information campaigns initiated after the reports on smoking and health from the U.S. Surgeon General and the Norwegian Board of Health in 1964. This article reports a study of differences in tobacco advertising in Norway before and after 1964. MATERIAL AND METHODS: A total of 1,945 photographs of advertisements in two popular weekly family magazines over the 1955 to 1975 period were stored in a database along with information on date of publication, size, copy and message, presumed target group, and characteristics of persons and situations shown. RESULT: Up until 1964, advertisement space per year in these magazines totalled 100 dm2 (11 sq. feet). The amount of advertising space then increased up until 1973, to 2,033 dm2 (219 sq. feet). The share featuring women went up from 33 in the 1955-64 period to 62% in 1965-75, when 51% of advertisements showed women smoking while 31% showed men smoking. Pre-1964 advertisements primarily contained information to smokers on price, type of tobacco, packaging and country of origin; after 1964 the advertisements developed a more universal appeal associating smoking with various social situations marked by style, well-being and comfort. The tobacco was said to be pure, fresh, mild and natural, and the filter was claimed to have protective properties. INTERPRETATION: It is probable that the cognitive dissonance created by more health information on smoking after 1964, i.e. the necessary motivation to quit, was reduced as a result of strategic changes in the amount and content of advertising. PMID- 11894606 TI - [Assisted insemination--a risk factor in perinatal health care]. PMID- 11894607 TI - [Asthma and compliance--is self-care possible?]. PMID- 11894608 TI - [Carisoprodol--one more time]. PMID- 11894609 TI - [Half of the Norwegian applications concerning specialist approval on Iceland are rejected]. PMID- 11894610 TI - [Most of the terminally ill patients want to die at home. Advanced home care services can often replace palliative hospital care]. PMID- 11894611 TI - [Well-founded guidelines must be updated. Continuous updating does not relieve the physician from the obligation to reflect]. PMID- 11894612 TI - [Risk and odds--how to calculate with events]. AB - Categorical scientific data consisting of counts of events are frequently reported as risk or odds figures. Given a certain set of data, odds always differs upwards from risk. The relation between two risk figures may be expressed in various ways, one of which is the risk ratio. In the case of two odds figures, the choice is almost always the odds ratio. With a certain set of data from two groups, the odds ratio is not identical to the risk ratio (except when both are equal to 1). The odds ratio always magnifies the intergroup difference. When assessing published data, one must take care to observe whether reported ratio figures denote risk ratios or odds ratios. PMID- 11894613 TI - [Communication problems in evidence-based medicine]. AB - From a humanistic, social scientific perspective, the most complex task in evidence-based medicine lies in the communication of specialized medical knowledge to non-professionals. Information is never simply the neutral transmission of facts, not even when dealing with scientific knowledge and research. It is always interpreted and evaluated from a particular perspective in a specific context. That information can be neutral is thus a myth. In all medical consultations the process of communication is not just a matter of transmitting information from one who knows to one who does not. Knowledge created and formulated in a scientific context is thus recontextualised first in a clinical situation and then as an interpreted version in people's real lives. Furthermore there are difficulties when practice must be based on current research, in a situation in which no prior clinical experience exists and in which results are interpreted and used regardless of the relative certainty of current evidence. PMID- 11894614 TI - [Palliative cancer care is an important part of urology. A study of the last year of life of patients who died because of urologic cancer]. AB - A total of 330 in-patient episodes at the urology ward, with a mean duration of 9.8 days, were registered in a study of 100 patients who died from urological cancer. Twelve patients spent more than two months of their last year of life at the urology ward. As many as 82% of the admittances were on an emergency basis. A total of 101 operations were performed on 84 patients; 47 patients received palliative radiotherapy. This patient category needs a great deal of palliative care--at short notice--in order to get an optimal quality of life. Although many symptoms could have been alleviated outside hospital, the majority of patients needed specialised urological hospital care during their last year of life. PMID- 11894615 TI - [A study of 20 general practitioners' role in the care of patients with cancer in Uppsala. Detailed specialist information facilitates the determination of support needs]. AB - The aim of this study is to describe the role of the GP in the care of one specified cancer patient per GP and to explore the GP's knowledge about that patient's disease and treatments. A further aim was to evaluate the effects of an extended information routine, including increased information from the specialist clinic to the GP. Twenty GPs were selected for a semi-structured interview about a patient randomised either to an extended GP information routine or to standard information. The results suggest that GPs are commonly involved in the care of cancer patients, particularly in the diagnosis of the disease but also during the period of treatments and follow-up. The information from the specialist clinic to the GP is insufficient in standard care. The extended information routine increased the GPs' knowledge about the disease and treatments and facilitated their possibilities to determine patients' need for support. However, this did not affect the extent of contacts with the patient. PMID- 11894617 TI - [Clinical investigation of cognitive dysfunction by specialists]. AB - The clinical investigation of the cognitively impaired, or demented, patient includes a clinical bedside examination supported by, for instance, structural and functional brain imaging, analyses of biochemical markers, and genetic analyses. Valuable discussions have been held during the last decade between specialists and primary care physicians to define the clinical elements that should be included in bedside examinations carried out by general practitioners. The present paper is focused instead on the bedside investigation performed by the specialist in the field. Emphasis is put on the importance of combined cognitive and neurological assessments to determine symptoms and syndromes that possibly reflect regional pathological changes in the brain. The article also stresses the importance of detecting possible behavioural and psychological signs of dementia, and the importance of collecting information on the premorbid personality of the patient. PMID- 11894616 TI - [Congenital dislocation of the knee in children--unusual condition of unknown etiology]. AB - Congenital dislocation of the knee is a rare condition. It results in an inability to flex the knee. The condition is normally diagnosed clinically during the neonatal period and is verified by X-ray and/or ultrasound. Treatment is preferably non-operative with gradually increased passive flexion of the knee. In certain cases surgery may be justified. In a majority of cases, the children achieve acceptable knee-function after treatment. In the region of Sundsvall Harnosand in Sweden, during the past seven years five children with congenital dislocation of the knee were born, which is a considerable accumulation of cases. This article describes these five children, their condition, possible etiology and strategies of treatment. PMID- 11894618 TI - [Living with dystonia. A questionnaire study among members of the Swedish Dystonia Patient Association]. AB - The dystonias are movement disorders with sustained muscle contractions causing twisting movements or abnormal postures. A patient survey sent to 268 members of the Swedish Dystonia Patient Association was returned by 221 (82%). Most of the patients (81%) were women and the most common form was cervical dystonia (spasmodic torticollis). Median age at onset was 41.6 years. The time from onset of symptoms to correct diagnosis was on average 6 years. 21% had relatives with movement derangements. Injection with Botulinum toxin and physical therapy were ranked as the most effective treatments. Many patients expressed a need for more physical therapy as well as a desire to meet the same physician each time and to receive more intensive psychological support. PMID- 11894619 TI - [Consensus on first-hand treatment of gastrointestinal lymphomas does not exist]. PMID- 11894620 TI - [Rehabilitation in temperate climate for persons with pelvospondylitis. Accessed disease activity and general well-being is gradually improving]. AB - The aim of the study was to investigate the effects on disease activity, range of motion, functional ability, and global wellbeing on persons with Ankylosing Spondylitis (AS) during three weeks' rehabilitation in a tempered climate. Forty eight persons were evaluated twice prior to the rehabilitation and three times after. The results indicated significant improvements on all variables assessed, which lasted three weeks and, in some cases, three months. Global wellbeing continued to improve during follow-up. The results of our study thus indicate that rehabilitation in a tempered climate might be a useful complement to traditional rehabilitation of persons with AS. PMID- 11894621 TI - [Comprehensive quality indicators for health care services]. AB - On behalf of the Swedish government, the National Board of Health and Welfare recently issued a recommended set of 60 quality indicators for broad monitoring of the quality of national health care. This initiative is in concordance with similar international initiatives. The Swedish process, however, is unique in the sense that the professions have developed the indicator and more than 40 national quality registers already monitor most of them. Health care professionals in Sweden have a long-standing tradition of measuring and monitoring results including comparing the quality of different health providers by means of the quality registers. However, the transparency by which these measures are presented, the general understanding of how such data should be interpreted and used in practice is as yet not sufficiently developed. Transparency of data combined with knowledgeable interpretation by health professionals will provide patients with sound information about health care quality and the necessary prerequisites for making comparisons between providers. It will also help guiding managers and politicians making decisions. In order to reach this objective, close co-operation between patient organisations, health care managers and the professionals is needed including a common understanding of the needs and perspectives of all parties. PMID- 11894622 TI - [Medical quality revision within psychiatry. A process which strengthens cooperation]. PMID- 11894623 TI - [How cure the sick health services?]. PMID- 11894624 TI - [Somatization--often a misleading explanation of symptoms difficult to understand]. PMID- 11894625 TI - [Watch out for the somatization trap!]. PMID- 11894626 TI - [Something is missing! Cultural diversity does affect the care]. PMID- 11894627 TI - [Private health services are strongly increasing in Sweden--but naturally the politicians must have some influence]. PMID- 11894629 TI - December 1901: 100 years ago La Medicina del Lavoro issued its first number. Introduction. PMID- 11894628 TI - [Folate and cobalamin in primary health care--time for a symposium!]. PMID- 11894630 TI - Pharmacologic treatment of inner ear: from basic science to the patient. AB - Most of the deafness are of sensorineural origin and are characterized by a loss of hair cells and of spiral ganglion neurons. At the present time, hearing aids are the only treatment. However, in some diseases of the inner ear, pharmacological treatment have been proposed and used successfully. In this paper, we will review some basic science aspects of the biology of the neurosensory structures of the inner ear, in particular of the auditory neurons, that lead to the rationale of some treatments for the inner ear diseases. Developmental studies, neuronal cell culture experiments, and analyses of gene knockout animals reveal a number of growth factors which are important for the rescue and repair of injured auditory neurons in the inner ear. These factors rescue the injured auditory neurons in vivo. Furthermore, perfusion of antioxydant to the cochlea prevented the hearing loss induced by cisplatin. These in vitro and in vivo experiments demonstrate that it is possible to manipulate the neurosensory structures of the inner ear and provide an effective treatment to prevent the degeneration of the neurons. The molecules or drugs can be administered locally to the inner ear through a direct perilymphatic perfusion or through the round window membrane. As an example, we will discuss the treatment of patients suffering from idiopathic sensorineural hearing loss which can be treated successfully by a perfusion through the round window membrane, improving their hearing threshold and their speech discrimination. PMID- 11894631 TI - Present state and future perspectives of computer aided surgery in the field of ENT and skull base. AB - BACKGROUND: CAS technology has emerged in the last 10 years and is more and more used for surgery of the anterior and lateral skull base. Endoscopic and microscopic CAS systems are available. The endoscopic key hole or minimal invasive procedure is used increasingly not only for treatment of inflammatory disease, but also for tumour surgery. The integration of CAS systems into these procedures raises the level of their safety and efficiency. In addition, they allow the distance to the bone border on the CT slices to be previewed and measured. OBJECTIVE: Review of the present state of CAS systems, our own experience as a research centre and as users of the systems, demonstration of new navigational devices, microscope integration and a new registration procedure developed for the "Bernese" frameless optical navigation system (SurgiGATE ORL@1000, medivision, [Synthes, Stratec-medical], CH-4436 Oberdorf). MATERIAL AND METHODS: The optical CAS system with microscope integration (VM 900@1000, Moller Wedel, Germany), new devices and new matching procedure; accuracy tests on a cadaver skull and on the patient's head. RESULTS: The practical accuracy on the cadaver skull with the pointer system has a mean average error of 0.79 mm, and the clinical accuracy is between 0.5 mm and 2 mm. CONCLUSION: CAS-microscope integration, new CAS technical devices and new noninvasive CAS-registration procedures support the surgeon in more minimally invasive surgical procedures of the paranasal sinuses and the skull base. Future perspectives will be discussed. PMID- 11894632 TI - Carcinoembryonic antigen in chronic otitis media: an immunohistochemical study of the middle ear mucosa. AB - Carcinoembryonic antigen (CEA) has an important role as a tumor marker and also as a cell adhesion molecule. The expression and the role of CEA in chronic inflammation of middle ear mucosa has not been studied previously. Immunohistochemical analysis with monoclonal antibodies against human CEA was performed in order to detect CEA in formalin fixed and paraffin embedded biopsy specimens of middle ear mucosa in chronic otitis media. In this study 118 specimens of middle ear mucosa from different regions of the middle ear cleft were analysed. All specimens were taken during surgery of otitis media with and without cholesteatoma. The results showed that CEA expression was present in mucosal samples in chronic otitis media with cholesteatoma, predominantly in squamous epithelium, as well as in the subepithelial layer and among the connective tissue, especially on collagen fibres of fibroproliferative stroma. CEA was seen on endothelial cells of blood vessels. Occasionally it was present on the surface of columnar epithelium of the middle ear mucosa in chronic otitis media without cholesteatoma. The expression patterns of CEA in chronic inflammation of middle ear mucosa suggest potential functional activities similar to adhesion molecules and signal regulatory proteins which will be the subject of further study. PMID- 11894633 TI - Abnormal petrous apex aeration. Review of 12 cases. AB - OBJECTIVE: Abnormal aeration of the petrous bone, which may be responsible for some clinical symptoms, is an incidental radiological finding. It has been reported that people with abnormal aeration at the petrous bone are more prone to develop complications. In an attempt to make clear the clinical importance of a giant air cell at the petrous apex or abnormal aeration of this region, a prospective analysis has been planned. METHOD: 430 temporal bone CTs taken for diagnostic imaging only between 1992-2000 have been reviewed and 12 cases with petrous air cell were selected. Aeration was measured on computer basis. Patients were invited for interview and subjected to complete audilogical and ENG testing Symptoms of the patients were reviewed and the possible link with the radiological findings has been discussed. RESULTS: The internal distance of the air cell was ranging between 1.6 to 2.5 cm at transverse axis and between 2.0 to 4.2 cm at longitudinal axis. It was noted that all patients with large air cell at the apex also had extensive mastoid and temporal bone aeration. The degree of temporal bone aeration and the size of air cell was the same in both sexes. Four patients had balance problems. One patient had sudden hearing loss. Two patients had Bell's palsy. Two patients were asymptomatic. Six patients had normal hearing level. Five patients demonstrated ENG abnormality. CONCLUSIONS: It was found that these patients had some ear symptoms which were usually vague and nonlocalizing. It was unlikely to attribute these symptoms to a giant apical air cell or abnormal aeration of the petrous bone. PMID- 11894634 TI - Comparison between caloric and canal impulse rotatory test. AB - It has been suggested that the rotatory test using videosnystagmoscopy can also be applied for identification of vestibular canal paresis. In this study, we test this hypothesis by comparing the results of a canal impulse rotatory test with those of the caloric test using the method described by Freyss. Our study indicates that only in 38% of all tested patients, the same results between the two tests have been obtained. Furthermore, which the group with abnormal findings is observed separately, in 24% patients only these two test methods lead to the same results. Consequently, the canal impulse rotatory test cannot replace the caloric test. PMID- 11894635 TI - Unknown primary detected by FDG-PET. A review of the present indications of FDG PET in head and neck cancers. AB - To investigate the indications of 18F-2-fluoro-2-Deoxy-D-glucose (FDG) Positron Emission Tomography (PET) in head and neck cancer, the present study focuses on a case of cervical metastatic lymph node, which is not detected by the routine approach. It discusses the potential role of FDG-PET in the detection of unknown primary tumors, lymph node metastasis and post radiation follow-up, and demonstrates the implications of its findings through a few examples. Based on the literature in the field of head and neck oncology, the paper recommends the following uses for FDG-PET: 1. To guide biopsy or even local resection at the initial stage of examining the unknown primary lesions in case of high clinical suspicion 2. A whole body PET in high-risk patients may prevent unnecessary treatment and reduce the number of examinations 3. To monitor tumor response before full-dose irradiation so as not to delay the salvage surgery when applicable 4. To detect residual, recurrent or secondary neoplasm after definitive radiotherapy at least 4 months post-treatment 5. To revise the necessity of neck treatment in case of a negative PET, in the NO necks; and 6. In cases of clinical suspicion for laryngeal cancer recurrence and absence of objective findings before obtaining biopsy. PMID- 11894636 TI - Subglottic hemangioma treated with interferon alpha 2A. AB - Infantile subglottic hemagioma is a rare vascular malformation involving the subglottic larynx and although present from birth, symptoms will not be noted until later in infancy (due to tendency to enlarge). Typically presents with a progressive crouplike illness that begins a few weeks after birth and the infant develops inspiratory stridor, which becomes expiratory as the obstruction increases. Although benign lesions which involute spontaneously, they may demand the attention of an ENT surgeon to maintain a secure airway. We report a case of a 3-month-old female patient (full term, normal delivery) who was referred to us for investigation of respiratory distress. Endoscopic examination revealed a hemangioma at the left lateral wall of the subglottic larynx while the rest of the airway was normal. The infant was treated with a combination of steroids (dexamethazone 0.5 mg x 3 per os) and interferon A-2a (650.000 IU subcutaneously, every other day, for 12 months) and had fast improvement of her symptoms. Repeated endoscopy 3 months after the diagnosis reveal impressive remission of the subglottic hemangioma. Throughout the years, a variety of treatments have been proposed and utilised for subglottic hemangioma. Interfron 2-alpha, drug acting by interference with angiogenesis, is very effective in treating subglottic hemangiomas without the need for tracheostomy. Its side effects are generally not serious. As congenital subglottic hemangiomas have tendency towards spontaneous regression, conservative treatment seems more appropriate compared to more aggressive treatment that carry substantial risks of long term complications. PMID- 11894637 TI - Deans ponder skills drain. PMID- 11894638 TI - Mozambique cholera will affect region. PMID- 11894639 TI - Long-term strategy to retain local nurses. PMID- 11894640 TI - Chickenpox vaccine snapped up instantly. PMID- 11894641 TI - State AIDS training steps up a gear. PMID- 11894642 TI - Ethical human rights and guidelines on HIV. II. PMID- 11894643 TI - Mother-to-fetus HIV transmission during amniocentesis--ethical concerns. PMID- 11894644 TI - The growing burden of heart failure. PMID- 11894645 TI - Monitoring of research--is it practical or only a dream? PMID- 11894646 TI - After the CBE. PMID- 11894647 TI - Recommendations pertaining to the use of viral vaccines: influenza. PMID- 11894648 TI - World Health Organisation 'healthy life expectancy in 191 countries, 1999'--what of the future? PMID- 11894649 TI - Assessing diagnostic accuracy and tympanocentesis skills of South African physicians in management of otitis media. PMID- 11894650 TI - An unusual case of pulmonary oedema. PMID- 11894651 TI - Risk factors for near-fatal asthma--a case-control study in a Western Cape teaching hospital. AB - BACKGROUND AND OBJECTIVES: To evaluate risk factors for asthma death such as access to health care, over-use of beta 2-agonists or under-use of inhaled corticosteroids in the Western Cape (WC) population, using near-fatal asthma (NFA) as a surrogate marker. SUBJECTS AND METHODS: Patients with NFA (cases) admitted to a WC teaching hospital were compared with patients with acute asthma in a case-control study using a structured questionnaire, clinical examination, arterial blood gas measurements, chest radiograph and pulmonary function measurements. RESULTS: Sixteen patients with NFA (cases) and 55 with acute asthma (controls) were prospectively enrolled. Duration of asthma, gender, smoking status and ethnicity were similar. Cases had significantly more previous mechanical ventilation (P < 0.05) and a trend towards more previous intensive care unit (ICU) admissions. No significant differences were found in primary health care variables. CONCLUSION: Our study demonstrates that patients with NFA constitute a significant number of emergency room (ER) admissions for acute asthma (30%) in our population. Similar to other studies, there was a trend for NFA toward more previous ICU admissions and mechanical ventilation. Relative under-use of beta 2-agonists the day before admission and fewer ER visits during the previous year in the NFA group, suggests an impaired perception of the severity of disease or a more rapid onset of symptoms. Negative factors such as inability to access health care or lack of medication supply were similar in both groups. The challenge remains to identify and manage high-risk patients effectively. PMID- 11894652 TI - What is wrong with my patient? How to read an article concerning diagnosis. PMID- 11894653 TI - Prevalence of pre-cancerous lesions and cervical cancer in South Africa--a multicentre study. AB - OBJECTIVES: To describe the age-specific prevalence rates of cancer of the cervix in South African women presenting for screening. DESIGN: A multicentre prevalence survey in 10 geographically defined areas following a common core protocol. Services were located in existing service sites, with the exception of KwaZulu Natal which used a mobile service. Women aged 20 years and above were eligible for inclusion. OUTCOME MEASURES: Age-specific cervical cytologically diagnosed abnormality rates according to the Bethesda classification. RESULTS: During the study 20,603 women participated. Eighty per cent of the sample had never had a Pap smear before and just over 91% had not had a Pap smear in the last 5 years. In this study population 468 women screened (2.42%) were found to have low-grade squamous intra-epithelial lesions (LSIL) and the average age of these women was 33.1 years; 366 (1.8%) had high-grade SIL (HSIL) and these women were statistically significantly older at 37.97 years of age; and 92 women (0.47%) were found to have cytologically diagnosed invasive cancer. These women were significantly older, with an average age of 51.3 years. A clear relationship was found between age and LSIL, with younger women having a high rate of LSIL which decreases with increasing age. A similar but inverse relationship between age and invasive cancer is described, with the rate being low in young women and increasing with increasing age. A clear relationship between HSIL and age is not described in these data. The adequacy rate (satisfactory and satisfactory but limited) of the slides was 95%, and just under 92% of the study sample received their results. Not all women were appropriately referred and it was not possible to assess if women referred for treatment received it. CONCLUSIONS: These data indicate that cancer of the cervix is a common disease and that, similar to other countries, it is a disease of older women. These data give some positive indicators for future screening--older women will present for screening and the majority of women received their results. However, improvements in health system functioning are needed. A uniform national cytology reporting system is required as well as clear guidelines for providers on what action to take based on cytology reports. Linkage between the site of screening and treatment centre is inadequate and requires urgent attention in order to decrease cervical cancer mortality. PMID- 11894654 TI - Staff/bed and staff/patient ratios in South African public sector mental health services. AB - OBJECTIVES: To document staff/bed and staff/patient ratios in public sector mental health services in South Africa. DESIGN: Cross-sectional survey. METHOD: A questionnaire was distributed to provincial mental health co-ordinators requesting numbers of full-time equivalent (FTE) staff who provide mental health care at all service levels; numbers of psychiatric beds; and numbers of patients who attend outpatient departments, clinics and community health centres. The information was supplemented by consultations with mental health co-ordinators in each of the nine provinces. RESULTS: The staff/bed ratio for the country as a whole was 0.3 staff per bed. For the provinces, the staff/bed ratios were as follows: Eastern Cape 0.30, Free State 0.50, Gauteng 0.22, KwaZulu-Natal 0.34, Mpumalanga 0.89, North-West 0.27, Northern Cape 0.26, Northern Province 0.26, and Western Cape 0.59. For the country as a whole, the staff/bed ratios for each category of staff were as follows: total nursing staff 0.25, occupational therapists 0.01, occupational therapy assistants 0.01, social workers 0.01, community health workers 0.00, psychologists 0.00, intern psychologists 0.00, psychiatrists 0.00, psychiatric registrars 0.01, and medical officers 0.00. The ratio of ambulatory psychiatric service staff to daily patient visits (DPV) for the country as a whole was 0.6. CONCLUSIONS: Staff/bed ratios in South African mental health care are low relative to developed countries. Staff/DPV ratios highlight both the need to develop ambulatory care personnel for mental health care, and problems associated with monitoring the delivery and utilisation of mental health services within an integrated health system at primary level. PMID- 11894655 TI - Staff/population ratios in South African public sector mental health services. AB - OBJECTIVE: To document existing staff/population ratios per 100,000 population in South African public sector mental health services. DESIGN: Cross-sectional survey. METHOD: A questionnaire was distributed to provincial mental health co ordinators requesting them to provide the number of full-time equivalent (FTE) staff responsible for mental health care at all service levels. These data were supplemented by consultations with mental health co-ordinators in each of the nine provinces. Population data were obtained from preliminary findings of the 1996 census. RESULTS: The overall staff/population ratio per 100,000 population was 19.5, with an interprovincial range of 5.7-31.5. The staff/population ratios per 100,000 population for selected personnel categories (with the interprovincial ranges in brackets) were as follows: total nursing staff 15.6 (4.4-28.4), occupational therapists 0.4 (0.1-0.8), occupational therapy assistants 0.5 (0.0-1.3), social workers 0.5 (0.1-0.9), community health workers 0.3 (0.0-1.0), psychologists 0.3 (0.0-0.7), intern psychologists 0.3 (0.0-0.7), psychiatrists 0.4 (0.1-0.8), psychiatric registrars 0.4 (0.0-1.2), medical officers 0.4 (0.2-1.3), pharmacists 0.2 (0.1-1.1), and pharmacy assistants 0.2 (0.0-0.6). CONCLUSIONS: Relative to international settings, there are low levels of mental health staff provision in South Africa, and there is a large amount of variability between provinces. There are considerable challenges in monitoring mental health staff resources within an integrated health service. PMID- 11894656 TI - The MRC HIV/AIDS mortality report--South Africa's 'Apocalypse Now'. PMID- 11894657 TI - Stirring up pathologists. PMID- 11894658 TI - Stirring up pathologist. PMID- 11894659 TI - Ethical issues in continuing professional development. PMID- 11894660 TI - The Jaipur paradigm. PMID- 11894661 TI - Proportional tertiary bed distribution. PMID- 11894662 TI - [Comparison of a sulfate-free polyethylene glycol (PEG) solution with a standard solution for colonoscopy preparation]. AB - In order to ameliorate the unpleasant salty taste of a standard PEG-electrolyte lavage solution and thus improving acceptance of colonoscopy cleansing, a new sulfate-free, low sodium PEG-solution has been developed as long as 10 years ago. Comparative studies regarding acceptance showed conflicting results. This study was intended to compare the standard solution L (Cololyt) with the sulfate-free solution S (Colo-Sol) on a prospective basis in a collective of average gastroenterology outpatients. The 144 patients who underwent colonoscopy ingested 1 liter of solutions L and S each on the evening before and another liter of the preferred solution (preference) in the morning of the day of examination. Moreover the taste of both solutions was rated (grading scale 0-10). Tolerance and efficacy were registered as well. Preference and thus acceptance presented very well-balanced. Solution L was preferred as frequent (by 50.7% of patients) as solution S (49.3%) which was originally claimed to be better tasting. The respective preferred solutions were rated equal with mean +/- SD values of 6.6 +/ 2.2 for L and 7.0 +/- 1.9 for S. Efficacy and tolerance did not differ from a clinical point of view. PMID- 11894663 TI - [Hepatitis B virus infection: diagnosis, clinical sequelae, therapy and prevention]. AB - This article gives an overview on the clinical epidemiology of hepatitis B in Switzerland. It considers structure of the hepatitis B virus, serologic diagnosis of hepatitis B and its prevention and treatment. The main conclusions are as follows: 1. Hepatitis B prophylaxis is available. Juveniles should be vaccinated before taking up sexual activity. Pregnant women should be tested and their offspring immunised actively and passively when there is evidence of infection. 2. In cases of acute hepatitis B contact persons should be tested and vaccinated where appropriate. Treatment is not indicated. 3. For the treatment of chronic hepatitis B interferons and lamivudine are currently available. Advantages and shortfalls of the different forms of treatment are discussed. PMID- 11894664 TI - [Why 2 ears?]. AB - Having two ears instead of just one is advantageous in several respects. The danger of deafness is decreased, the auditory field is enlarged, spatial perception is possible on the basis of binaural disparities. Auditory deprivation presents as loss of discrimination over time in the unaided ear of individuals with a unilateral hearing loss. Earedness and ear preference ensure the choice of the dominant ear for definite actions like eavesdropping, telephone or listen. PMID- 11894665 TI - [Painless jaundice. Patient: 46-year-old teacher. Amoxycillin/clavulanic acid induced cholestatic hepatitis]. PMID- 11894666 TI - [Tropical tick bite. 46-year-old chemist. Rickettsiosis, R. africae (African tick bite fever)]. PMID- 11894667 TI - [Asbestos and fibrous materials]. PMID- 11894668 TI - [Photomicrography and diffraction atlas]. PMID- 11894669 TI - [Crystalline free silica]. PMID- 11894670 TI - Dead and forsaken. PMID- 11894671 TI - Domestic disputes. Nearly every state prohibits same-sex marriage. But what are the rights of transsexuals who wed? PMID- 11894673 TI - To clip or to coil? Today patients with cerebral aneurysms face a difficult choice: brain surgery or a less proven alternative. PMID- 11894672 TI - They still don't get it. How can a church that judges so many faithful cover up its own offenses? PMID- 11894674 TI - Sins of the fathers. PMID- 11894675 TI - Bing Crosby had it right. PMID- 11894676 TI - Avoid the four perils of CRM. AB - Customer relationship management is one of the hottest management tools today. But more than half of all CRM initiatives fail to produce the anticipated results. Why? And what can companies do to reverse that negative trend? The authors--three senior Bain consultants--have spent the past ten years analyzing customer-loyalty initiatives, both successful and unsuccessful, at more than 200 companies in a wide range of industries. They've found that CRM backfires in part because executives don't understand what they are implementing, let alone how much it will cost or how long it will take. The authors' research unveiled four common pitfalls that managers stumble into when trying to implement CRM. Each pitfall is a consequence of a single flawed assumption--that CRM is software that will automatically manage customer relationships. It isn't. Rather, CRM is the creation of customer strategies and processes to build customer loyalty, which are then supported by the technology. This article looks at best practices in CRM at several companies, including the New York Times Company, Square D, GE Capital, Grand Expeditions, and BMC Software. It provides an intellectual framework for any company that wants to start a CRM program or turn around a failing one. PMID- 11894677 TI - Getting the truth into workplace surveys. AB - There's no doubt that companies can benefit from workplace surveys and questionnaires. Good surveys accurately home in on the problems the company wants information about. They are designed so that as many people as possible actually respond. And good survey design ensures that the spectrum of responses is unbiased. In this article, the author, a former research scientist at the University of Michigan and currently the president of a survey design firm, explores some glaring failures of survey design and provides 16 guidelines to improve workplace assessment tools. Applied judiciously, these rules will not only make a tangible difference in the quality and usefulness of the data obtained but will also produce an increased response rate. The guidelines--and the problems they address--fall into five areas: content, format, language, measurement, and administration. Here are a few examples: Survey questions should require people to assess observable behavior rather than make inferences; each section should contain a similar number of items and each item should have a similar number of words; words with strong associations to gender, race, or ethnicity should be avoided; the wording in one-third of the questions should be changed so that the desirable answer is a negative one; and response scales should provide a "don't know" or "not applicable" option. Following the guidelines in this article will help you get unbiased, representative, and useful information from your workplace survey. PMID- 11894678 TI - Managing emotional fallout. Parting remarks from America's top psychiatrist. Interview by Diane Coutu. AB - Last fall, the United States was brutally thrust into a new and dangerous world. As the twin towers of the World Trade Center collapsed and the Pentagon burned, the horrible reality of terrorism seared the American consciousness. It touched more than the victims and their families; everyone who sat transfixed before the horrific images on TV lived through the trauma. In a sense, we were all eyewitnesses, and we must all cope with feelings of anger, stress, and anxiety. That poses a huge immediate challenge for business, because it is largely in the workplace--where we spend so many of our waking hours--that we will confront these emotions. And many companies have risen to the challenge, establishing new guidelines for processing mail in light of anthrax fears and organizing stress reduction programs for employees. While the logic of taking such action is incontestable, it raises a much larger question: What responsibility does a company bear for the mental well-being of its work-force? If companies help employees deal with depression and anxiety in the wake of terrorist acts, doesn't that put mental health care on the business agenda? To answer these questions, HBR senior editor Diane Coutu talked with Dr. Steven Hyman, the former director of the National Institute for Mental Health. In this interview, he discusses the implications of coping with tragedy, the resilience of individuals, and the treatment of mental illness. And he suggests that September 11, 2001, may come to be seen as a tipping point--the moment when managers started to think about dealing with mental health issues on a regular basis. PMID- 11894679 TI - Beware the busy manager. AB - Managers will tell you that the resource they lack most is time. If you watch them, you'll see them rushing from meeting to meeting, checking their e-mail constantly, fighting fires--an astonishing amount of fast-moving activity that allows almost no time for reflection. Managers think they are attending to important matters, but they're really just spinning their wheels. For the past ten years, the authors have studied the behavior of busy managers, and their findings should frighten you: Fully 90% of managers squander their time in all sorts of ineffective activities. A mere 10% of managers spend their time in a committed, purposeful, and reflective manner. Effective action relies on a combination of two traits: focus--the ability to zero in on a goal and see the task through to completion--and energy--the vigor that comes from intense personal commitment. Focus without energy devolves into listless execution or leads to burnout. Energy without focus dissipates into aimless busyness or wasteful failures. Plotting these two traits into a matrix provides a useful framework for understanding productivity levels of different managers. Managers who suffer from low levels of both energy and focus are the procrastinators: They dutifully perform routine tasks but fail to take initiatve. Disengaged managers have high focus but low energy: They have reservations about the jobs they are asked to do, so they approach them half-heartedly. Distracted managers have high energy but low focus: They confuse frenetic activity with constructive action. Purposeful managers are both highly energetic and highly focused: These are the managers who accomplish the most. This article will help you identify which managers in your organization are making a real difference--and which just look busy. PMID- 11894680 TI - They're not employees, they're people. AB - In this essay, business thinker Peter Drucker examines the changing dynamics of the workforce--in particular, the need for organizations to take just as much care and responsibility when managing temporary and contract workers as they do with their traditional employees. Two fast-growing trends are demanding that business leaders pay more attention to employee relations, Drucker says. First is the rise of the temporary, or contract, workers; 8 million to 10 million temp workers are placed each day worldwide. And they're not just filling in at reception desks. Today, there are temp suppliers for every kind of job, all the way up to CEO. Second, a growing number of businesses are outsourcing their employee relations to professional employee organizations (PEOs)--third-party groups that handle the ever mounting administrative tasks associated with managing a company's employees. (Managers can easily spend up to one-quarter of their time on employee-related rules, regulations, and paperwork.) Driving these trends, Drucker observes, is the shift from a dependency on manual labor to create wealth and jobs to a dependency on specialization and knowledge. Leaders are increasingly trying to keep up with the needs of many small groups of product or service experts within their companies. Temps and PEOs free up leaders to focus on the business rather than on HR files and paperwork. But if organizations outsource those functions, they need to be careful not to damage relationship with their people in the process, Drucker concludes. After all, developing talent is business's most important task--the sine qua non of competition in a knowledge economy. PMID- 11894681 TI - Are you picking the right leaders? AB - When it comes time to hire or promote, top executives routinely overvalue certain skills and traits while overlooking others. Intuitively, for example, they might seek out team players, people who shine operationally, dynamic public speakers, or those who are demonstrably hungry for greater responsibility. But some attributes that seem like good indicators of leadership potential are, paradoxically, just the reverse. Team players and those who excel operationally often make better seconds in command. Many a great public speaker lacks the subtle one-on-one persuasive powers that a top leader needs. And shows of raw ambition may be more an indicator of ego than of leadership talent. Unfortunately, few organizations have the right procedures in place to produce complete and accurate pictures of their top prospects. Assessments are often based on hearsay, gossip, and casual observation. Many companies spend too much effort trying to develop leaders and not enough effort trying to identify them. A new evaluation process will help you avoid that trap. Candidates are assessed by a group of people who have observed their behavior directly over time and in different circumstances. Using a carefully crafted series of questions, the group can probe a wide range of leadership criteria, including such "soft" attributes as personal integrity, that are difficult to assess. Without such information, senior management will remain vulnerable to misidentifying leadership talent, and the wrong people will continue to make their way up the corporate ladder. PMID- 11894682 TI - A brief introduction to identifying a "good" qualitative study when you see one. PMID- 11894683 TI - Weaving for the future: using rulers and roses. PMID- 11894684 TI - Cancer nursing: weaving the tapestry for our second century. PMID- 11894685 TI - Evaluation of a breast self-examination (BSE) program in a breast diagnostic clinic. AB - The purpose of this study was to evaluate the short-term effectiveness of a breast self-examination (BSE) teaching program on women's knowledge about BSE, proficiency in performing BSE, and motivation to perform BSE. The program was developed for delivery by nurses in a breast diagnostic clinic, a clinic designed to meet the need for expeditious management of breast disease, current information about breast cancer risk, surveillance, and counselling. A convenience sample of 68 women attending the clinic in a regional cancer centre participated in a pre- and five month post-teaching program evaluation. The Toronto Breast Self Examination Instrument was used as the evaluation tool. There were statistically significant changes following the teaching program in the areas of knowledge about the correct technique for performing BSE, proficiency performing BSE, and confidence about finding changes when performing BSE. No significant changes were observed in motivation to practise BSE, although group scores did improve following the education. Participants found the video presentation and the review of BSE information pamphlets by the nurse to be the most helpful components of the BSE teaching program. PMID- 11894686 TI - A comprehensive review of selected cancer websites. AB - With increasing availability of information on the Internet, it makes sense for health care professionals to use this resource to expand the range of resources available to both themselves and to clients. This paper was designed to provide health care professionals with a starting point from which to gather further website oncology resources. An established template for evaluating websites is described and criteria regarding the accuracy of information, site design, and navigation are examined. While the initial phase of this paper evaluated numerous sites, only the top five as noted by the author are presented here. PMID- 11894687 TI - Redesigning processes in ambulatory chemotherapy: creating a patient appointment scheduling system: Part II. PMID- 11894688 TI - Experiences of women with recurrent ovarian cancer. PMID- 11894689 TI - [Abdominal compartment syndrome: a complication still unknown]. PMID- 11894690 TI - [Benign hepatic tumors: diagnostic management and therapeutic outcome]. AB - The number of non-malignant tumors of the liver diagnosis has greatly increased with widespread use of abdominal ultrasonography. Unlike hemangioma and focal nodular hyperplasia does not require surgery as does hepatocellular adenoma due to the risk of fatal complications. Differentiation between these three kinds of benign tumors is of prime importance to avoid unnecessary surgical interventions. Progress in medical imaging facilitates identification of hemangioma and focal nodular hyperplasia, but in case of doubt, histological proof must be obtained. Laparoscopy may be indicated not only for diagnostic but also for therapeutic purposes. PMID- 11894691 TI - [Abdominal compartment syndrome]. AB - Abdominal compartment syndrome (ACS) is defined by the deleterious effects of intraabdominal hypertension (IAH) on the pulmonary, cardiovascular, splanchnic, urinary and central nervous system. Abnormal and sudden increase in the volume of any component of the intraperitoneal or retroperitoneal space (occurRing postoperatively or subsequent to hemorrhagic trauma, referfusion edema, penumoperitoneum, intestinal distention, acute pancreatitis...) causes IAH. Sustained IAH leads to ACS which if left unrecognized or untreated is always fatal. Measurement of urinary bladder pressure is the best validated technique for diagnosis of IAH. It should be used routinely for minimally invasive surveillance of intra-abdominal pressure (IAP) in patients with severe thoraco abdominal trauma or after major abdominal operations. Medical management of IAH is of limited efficacy making expedient surgical decompression the treatment of choice for ACS. Surgical decompression of the abdomen and temporary closure is generally recognized as effective in clinically patent ACS but the pressure threshold indicating the need for decompression remains controversial. No data are available from controlled randomized trials and current guidelines are based on the experience of large trauma centers. The few available prospective clinical series report survival rates in the 38 to 71% range after surgical decompression for ACS. These studies are difficult to compare due to methodological features but it would appear that centers using the lowest pressure threshold for decompensation (< 20 mmHg) have the highest survival rates. Despite the available physiological arguments, indications for prophylactic temporary abdominal coverage (TAC), e.g. in trauma patients or for early decompression in IAH patients without clinical ACS, have not been validated in clinical practice. The potential morbidity of decompression procedures, TAC, and subsequent abdominal wall reconstructions require comparative studies of these treatment options with available pharmacological and non-surgical means to lower IAP. PMID- 11894692 TI - [Anal condyloma: its management is still difficult]. AB - Anal condylomas result from papillomavirus infection, the most common sexually transmitted disease. The principal risk is the development of cancer of the anal canal. The risk of contamination is high, even after a single sexual contact. No specific antiviral treatment is available and no consensus has been reached on the appropriate treatment of anal condylomas. Despite the development of various methods (interferon, imiquimod), electrocoagulation remains the treatment of choice. Finally, regular follow-up and treatment of the partner(s) are essential. PMID- 11894693 TI - [Anteriorly enlarged nephrectomy]. PMID- 11894694 TI - [Extra-mucosa pyloromyotomy for pyloric hypertrophic stenosis]. PMID- 11894695 TI - [Treatment of the body and tail of the pancreas after cephalic duodenopancreatectomy]. PMID- 11894696 TI - [Adrenal hemangioma]. PMID- 11894697 TI - [Hematoma of the abdominal large right muscles: diagnosis and treatment]. AB - Rectus sheath hematoma is an uncommon event, and exception post-trauma hematoma, affects predisposed patients. The typical clinical case combine abdominal pain, a palpable mass, and parietal eccymosis appearing when a patient under anticoagulant therapy coughs. Diagnosis is confirmed by abdominal ultrasonography or CT scan. Surgical treatment is indicated only in complicated forms. PMID- 11894698 TI - [A 61-year-old woman with jaundice after laparoscopic cholecystectomy]. PMID- 11894699 TI - [Public health--violence and sexual abuse]. PMID- 11894700 TI - [Liver cirrhosis mortality. Denmark in an international perspective--what direction are we taking?]. PMID- 11894701 TI - [Genetic defects in insulin signalling proteins. Implications for the pathogenesis of type 2 diabetes]. AB - The pathogenesis of type 2 diabetes mellitus is complex and involves abnormalities in both the action and secretion of insulin. These abnormalities are caused by a complicated interplay between genes and environment. A determination of the genetic defects that predispose to either insulin resistance or decreased insulin secretion is important, as an improved understanding of the underlying molecular mechanisms may be essential for the development of the most effective treatment. This paper focuses on genetic variants identified in genes encoding proteins in the early insulin signalling cascade. Variations frequently occur in these genes, but their effect varies in different populations. This may suggest that the genetic background is a considerable factor and that the synergistic effect of several variants plays a major role. Future, genetic epidemiological studies of large populations are therefore important in order to obtain sufficient statistical power. The paper also discusses recent results that suggests, that insulin itself has an effect on insulin secretion by the beta-cell and that insulin signalling in the CNS plays an important role in the regulation of energy disposal, fuel metabolism, and reproduction. PMID- 11894702 TI - [Magnetic resonance and multiple sclerosis I. Conventional diagnostic techniques]. AB - Conventional magnetic resonance imaging (MRI) is still the standard technique in the diagnosis and follow-up of patients with multiple sclerosis (MS), despite low pathological specificity and poor correlation to the disability. The results of T2- and T1-weighted imaging with and without contrast and the lesion characteristics are presented in this review. The numerous differential diagnoses and the diagnostic MRI criteria are also reviewed. PMID- 11894703 TI - [Magnetic resonance and multiple sclerosis II. New diagnostic techniques]. AB - Conventional magnetic resonance imaging (MRI) techniques have proved important in the diagnosis and in the follow-up in clinical trials of patients with multiple sclerosis (MS). However, these techniques have low specificity for the pathological changes in the MS lesions, and the correlation between conventional MRI and the disability is poor. The last ten years have seen the development of new techniques with improved sensitivity and increased pathological specificity, such as magnetisation transfer imaging (MTI), magnetic resonance spectroscopy (MRS), diffusion-weighted imaging, and functional magnetic resonance imaging (fMRI). These techniques and their contribution to the knowledge about the pathophysiology of MS are described in this review. PMID- 11894704 TI - [Mammographic screening in the county of Copenhagen. Clinical consequences of the first three screening rounds]. AB - INTRODUCTION: Service mammography has been offered biennially to women aged 50-69 years in the municipality of Copenhagen since 1991. The results were compared to breast cancer cases before initiation of screening. The comparison concerns prognostic factors and the treatment-related consequences. MATERIAL AND METHODS: Data from the Copenhagen service mammography screening were linked to data from the DBCG database. RESULTS: Before screening, 16% of breast cancer cases had a tumour size of 10 mm or less, this percentage increased to 41 in the screen detected cases. Sixty per cent of breast cancer cases showed no evidence of metastatic spread to axillary lymph nodes before screening; this percentage increased to 78 per cent in the screen-detected cases. Forty per cent of ductal carcinomas showed a malignancy grade I before screening, compared to 53% in the screen-detected cases. Thirteen per cent were treated with breast conserving therapy before screening, as opposed to 48% in the screen-detected cases. Forty one per cent needed postoperative adjuvant treatment before screening, compared with 21% in the screen-detected cases. DISCUSSION: A marked improvement was seen in the prognostic and treatment-related characteristics of the screen-detected breast cancer cases, as compared to breast cancer cases from the same area before screening was initiated. PMID- 11894705 TI - [Intensive physical therapy after trochanteric femoral fracture. A randomized clinical trial]. AB - INTRODUCTION: This randomised study evaluates the effect of intensive physical therapy on the duration of rehabilitation after hip fracture. MATERIALS AND METHODS: The study comprised 88 patients transferred for rehabilitation after operative treatment for hip fracture. After randomisation, 44 patients received physical therapy 3.6 hours (median) a week, whereas the 44 control patients received physical therapy 1.9 hours a week. The outcome was defined as the duration of physical rehabilitation until the patient was able to 1) walk 50 metres in less than two minutes; 2) manage to climb stairs to the first floor; 3) manage the sit-to-stand transfer; 4) get in and out of bed; 5) manage bathing, dressing, and lavatory visits. RESULTS: In the group randomised to intensive physical therapy, 24 patients dropped out after 15 days (median) whereas 13 patients dropped out of the control group after 22 days. Drop outs were caused by orthopaedic complications, general weakness, and poor co-operation. No difference was found in the duration of physical rehabilitation by analysis per protocol of the patients who completed the trial. DISCUSSION: The considerable number of drop outs suggests that intensive physical therapy may be of limited value in the attempt to reduce the duration of rehabilitation after hip fracture. An altered objective, including enhanced outpatient rehabilitation, may be necessary in order to reduce the length of hospital stay after hip fracture. PMID- 11894706 TI - [Violence in Aarhus over two decades]. AB - INTRODUCTION: Data collected from hospitals and the police have on several occasions proved to be useful in gaining knowledge about violence. The casualty departments, the Institute of Forensic Medicine, and the police in Aarhus have co operated in three earlier studies of inter-person violence, the latest in 1993 1994. The aim of this study was to update knowledge about the incidence and character of violence in Aarhus. MATERIAL AND METHODS: A dynamic cohort study was carried out in the period 1 April 1999 to 31 March 2000. Persons who arrived at the casualty wards or Institute of Forensic Medicine after having been exposed to violence were registered and interviewed. Data from the police were also collected. All results were compared to corresponding data collected in 1981 1982, 1987-1988, and 1993-1994. RESULTS: The number of victims decreased significantly to 1496. The annual incidence rate dropped to 4.6 victims/1000. This decrease in violence was particularly seen among 15-24-year-old males. The extent of violence against women remained unchanged in the years 1999-2000. Foreigners were significantly higher represented among the victims. The character and severity of the violence were unchanged. DISCUSSION: Similar findings with an overall decrease in the incidence of violence and, at the same time, an increase in violence among foreigners have been reported in studies from Norway and Britain. Preventive action should be considered in the groups of 15-24-year-old males and foreigners. PMID- 11894707 TI - [Mammography screening in the county of Copenhagen. Results of the first three screening rounds]. AB - INTRODUCTION: Biennial service mammography screening for breast cancer has been offered to women aged 50-69 years in the municipality of Copenhagen since 1991. We report the results of the first three invitation rounds. MATERIAL AND METHODS: Data were collected from the Copenhagen service mammography screening database and other Danish registers. RESULTS: The average participation rate during the first three invitation rounds was 66%. The breast cancer detection rate was 10/1,000 screened in the first invitation round and 5/1,000 in the consecutive rounds. The probability of a false positive mammography was 6% at the prevalent screen, and this was reduced to 3% at incidence screens. Fifty-two cases of interval cancer were seen after the first invitation round. The expected number was 152, which gives a proportional interval cancer rate of 0.34. The sensitivity was 86% and the specificity 94% after the first round. DISCUSSION: The detection rate of breast cancer was high, especially in the prevalence round. The trend in the incidence of breast cancer at the subsequent rounds was similar to that before screening, which indicates that mammography screening does not lead to any greater over-diagnosis. The rate of false positive mammography was high at the initial screening round, but was acceptable at subsequent rounds, and a false positive mammography does not seem to have affected participation in subsequent rounds. The Copenhagen screening programme conforms to international quality assurance guidelines for process evaluation. PMID- 11894708 TI - [Tuberculous spondylitis--Pott disease--in a 33-year-old man from Greenland]. AB - We report a case where tuberculous spondylitis was diagnosed in a 33-year-old man from Greenland. Very typically for Pott's disease, there was back pain, weight loss, and a characteristic kyphotic deformity. Computed tomography showed destruction of the thoracic vertebrae with paraspinal abscesses. The treatment was antituberculous therapy and surgery. PMID- 11894709 TI - [Pseudarthrosis of the ulna in a patient with neurofibromatosis]. AB - A case of pseudoarthrosis and partial resorption of the ulna in a 12-year-old girl with neurofibromatosis is described. Neurofibromatosis is a common neurocutaneous disease with characteristic osseous lesions. The pseudoarthrosis in this case developed after fracture. The bone most commonly affected is the tibia and involvement of the ulna has been reported in only a few cases. The possible cause of pseudoarthrosis, the radiographic changes, and the treatment are discussed. PMID- 11894710 TI - [Particulate air pollution and risk of ischemic heart disease]. PMID- 11894711 TI - [The Danish Society of Pain Medicine]. PMID- 11894712 TI - [Smoking and long-term oxygen therapy. Indications and contraindications]. PMID- 11894714 TI - [Function of the rheumatoid hand]. PMID- 11894713 TI - [Diagnosis and treatment of early rheumatoid arthritis]. PMID- 11894716 TI - From the chair: sharing our worth. PMID- 11894715 TI - [Fish oil]. PMID- 11894718 TI - From a past chair. Benchmarking implementation task force. PMID- 11894717 TI - Governmental relations. PMID- 11894719 TI - OPL. Myths and legends 101. AB - Libraries are the topic (and occasional font) or numerous myths and legends. Solos find themselves confronting any number of beliefs firmly held by their constituents as gospel-trye, in spite of evidence to the contrary. Librarians themselves are not immune to this condition. Herewith are some of my favorites, in no particular order. PMID- 11894720 TI - CHI services: consumer outreach coordinators at regional medical libraries ready to serve your professional needs! PMID- 11894722 TI - Diabetic nephropathy. Pathogenetic aspects and cardiovascular risk factors. PMID- 11894721 TI - Bisphosphonates for prevention of postmenopausal osteoporosis. AB - Our studies showed that 5 mg alendronate per day was the lowest, most effective dose that persistently prevented bone loss in recently postmenopausal women with normal bone mass. The effect on bone mass and biochemical markers was found comparable to that of commonly recommended regimens of postmenopausal HRT, and 5 mg alendronate per day is suggested as a new option for prevention of postmenopausal osteoporosis. HRT must, however, still be considered the first choice for this indication because of additional beneficial effects on other organ systems. The effect of alendronate was unaffected by bone or fat mass status, but increased with increasing postmenopausal age. The implications were that alendronate stabilized bone mass to a comparable extent in women at particular risk of osteoporosis because of thin body habitus or low bone mass and in healthy postmenopausal women with normal bone mass. Calcium supplementation was insufficient to prevent bone loss and did not add an effect on bone metabolism when combined with alendronate treatment in recently postmenopausal women. The gastrointestinal risk and adverse event profile of 5 mg alendronate per day was comparable to that of placebo, and this dose of alendronate appeared safe for long-term use. Bone loss resumed at a normal postmenopausal rate promptly after withdrawal of alendronate in early postmenopausal women consistent with a substantial underlying natural bone loss during early menopause. Oral ibandronate increased bone mass at all skeletal regions in elderly postmenopausal women with low bone mass, and 2.5 mg ibandronate per day was the lowest dose with this effect. The results are indicative of ibandronate as an option for secondary prevention of postmenopausal osteoporosis, but longer-term phase III trials should be performed before ibandronate can be recommended for this indication. The study showed that 2.5 mg ibandronate per day was efficient for prevention of bone loss and increment in bone mass in a population of women at particular risk of osteoporosis because of low bone mass. There were no differences between 2.5 mg ibandronate per day and placebo in terms of side effects, including complaints from the gastrointestinal tract, and ibandronate appeared safe for longer-term use in this dosing. Bone loss resumed at a normal postmenopausal rate when treatment was withdrawn. The response in bone mass and biochemical markers indicated that 2.5 mg ibandronate per day is equivalent to 10 mg alendronate per day in postmenopausal women. Our studies of two recently developed biochemical markers, urine CTX and serum total OC, showed that bone turnover was lowest in the premenopausal period, where these biochemical markers furthermore revealed a negative association with bone mass. It indicated that increased bone turnover contributes to a small premenopausal bone loss and resulting lowered bone mass. In consistence, a small premenopausal bone loss was observed in some regions of the hip. The biochemical markers increased at the time of menopause, consistent with initiation of the postmenopausal bone loss, and became gradually more negatively associated with bone mass as time past the menopause increased. The biochemical markers were furthermore higher in postmenopausal women with low bone mass, consistent with the characterization of postmenopausal osteoporosis as a condition with increased bone turnover. Our results consistently indicated a central role of increased bone turnover for development of low bone mass and osteoporosis. It is, however, also important to stress that the associations between biochemical markers and bone mass were too weak to allow for a valid individual estimation of bone mass based on biochemical markers. In contrast, the biochemical markers were shown as valid tools for monitoring and prediction of treatment effect of bisphosphonates. CTX, NTX, and total OC revealed the best performance characteristics in this respect. Six months after start of treatment, the level of suppression of these biochemical markers of bone resorption and formation accurately reflected the size of the 1-2 year response in bone mass in groups of women treated with bisphosphonate. This was a clear advance over bone densitometry, which has a precision error in the area of the anticipated yearly bone mass response during bisphosphonate therapy. The relationship was consistent during treatment with alendronate or ibandronate and in younger or elderly postmenopausal women. In individual patients, cut-off values of an about 40% decrease in urine CTX or NTX and an about 20% decrease in total OC validly predicted long-term prevention of bone loss. The sensitivity of prediction was high, but the specificity low. This implicated that the biochemical markers could be used as an exact method to detect "responders" to therapy, whereas "non responders" to bisphosphonate treatment should be detected with bone densitometry in patients who do not reveal a decrease below the cut-off value in the biochemical marker during treatment. However, before such approach can be generally recommended the cut-off values of the biochemical markers should be validated in future clinical trials of bisphosphonate. Postmenopausal osteoporosis develops slowly over many years and mainly becomes a significant individual and socio-economic health problem 1-3 decades after the menopause. Prevention of postmenopausal osteoporosis by bisphosphonates is therefore likely to imply a treatment regimen of at least a decade, as presently recommended for HRT (Consensus Development Statement 1997). However, future cost-effectiveness studies should reveal when bisphosphonate treatment should ideally be initiated. Our studies showed that the bisphosphonates were effective over the range from general recommendation (recently postmenopausal women with normal bone mass) to a reservation for women at particular risk of osteoporosis (elderly women, thin women, or women with osteopenia). Presently available biochemical markers could be used for groupwise and individual monitoring and prediction of treatment response. Most presently available biochemical markers, however, have the drawback of a low specificity. Recent studies of CTX measured in serum are promising, and indicate that this new biochemical marker might have overcome these drawbacks due to a pronounced response to treatment and a low long-term biological variation (Christgau et al. 1998b, Rosen et al. 1998, and 2000). PMID- 11894723 TI - Pharmacodynamic effects of oral contraceptive steroids on biochemical markers for arterial thrombosis. Studies in non-diabetic women and in women with insulin dependent diabetes mellitus. AB - Even small increases in the frequency of thrombotic disease in users of OCs have general health impact because of their widespread use, which is currently expanding to potential risk groups. The present investigations were launched to study the effects of OCs containing 20-40 micrograms of EE combined with the latest developed gonane progestogens on biochemical risk markers within metabolic systems involved in the development of arterial thrombotic disease. The studies included evaluation of carbohydrate and lipid metabolism as well as the haemostatic system and were performed in non-diabetic women and in women with IDDM, who are prone to the development of arterial thrombosis. In the evaluation of the carbohydrate metabolism in non-diabetic women, we found no effect on fasting glucose or insulin and no effect on the insulin response to oral glucose in women using monophasic OCs containing EE combined with DSG or GST. This contrasts the evaluation of triphasic OCs containing EE combined with GST or NGT, which increased fasting insulin and reduced insulin sensitivity without affecting the glucose-effectiveness or the beta-cell function. Impaired glucose tolerance developed in 10% of the women after 6 months. These finding suggest that OCs are able to induce a state of insulin resistance, which should be considered in the prescription for women with potential disturbed insulin sensitivity or reduced beta-cell secretory capacity e.g. women with ovarian hyperandrogenism, obesity, previous GDM or perimenopausal women. We found no change in glycaemic control in 22 women with well-regulated IDDM treated with a monophasic combination of EE and GST for one year and none of the women developed microalbuminuria during treatment. In the women with diabetes we observed an increase in fasting levels of triglycerides, a decrease in LDL-cholesterol, and unchanged concentrations of total cholesterol and HDL-cholesterol during treatment. In non-diabetic women treated with the same compound or an OC containing EE and DSG we found similar changes in triglycerides and total cholesterol, but increased levels of HDL cholesterol and unchanged LDL-cholesterol concentrations. In the women with IDDM there was a negative correlation between daily insulin requirement and HDL cholesterol before and during treatment, but no other statistically significant correlation between estimates of glycaemic control and lipids and lipoproteins were observed. In the non-diabetic women, changes in the haemostatic system included an increase in the procoagulant factors fibrinogen and Factor VIIc; the concentration of active t-PA increased, mainly because of decreased inhibition by PAI-1. The ratio between molecular markers of the activity of the coagulation system and the efficacy of fibrinolysis was unchanged. This was also found in the women with IDDM, who showed evidence of increased fibrin formation and an attenuated fibrinolytic response during treatment. The regulation of the t-PA/PAI system was studied in non-diabetic women in order to elucidate if the effects of OCs are caused by a direct effect on synthesis or clearance of these variables or if they are secondary to changed insulin sensitivity, as described in individuals with atherosclerosis. We found no indications that insulin resistance is involved in the regulation of t-PA and PAI-1 antigen levels, neither before nor during intake of OCs. We showed, however, that the decreased t-PA antigen concentration observed in OC users is caused by reduced synthesis outside the splanchnic circulation. The studies indicate that low-dose OCs containing newer gonane progestogens are able to induce insulin resistance and to impair glucose tolerance. Lipoproteins were not adversely influenced by the OCs neither in the diabetic nor the non-diabetic women; on the contrary, there was a tendency towards increased plasma levels of HDL-cholesterol and decreased LDL-cholesterol which are associated with a decreased risk of atherosclerosis. The changes observed within the haemostatic system were in accordance with a maintained balance between coagulation and fibrinolysis although the rate of fibrin formation may be increased in the women with IDDM. Irrespective of OC use, the interrelationships between metabolic systems in young non-diabetic women are different from those reported in individuals with atherosclerosis or insulin resistance. The effects of OCs on the t-PA/PAI system seem to be mediated by a direct effect on the vessel wall and not by changes in the hepatic clearance. The present findings were obtained in diabetic women without vascular complications, so the conclusion that women with IDDM can use OCs without metabolic alterations of known clinical significance is therefore restricted to those without evidence of diseased vessels. When evaluating the results obtained in the non-diabetic women, it should be remembered that women with recognised risk factors were excluded. The results may therefore be of limited value when evaluating the risk of arterial thrombosis in predisposed populations. In healthy individuals, the present integrated evaluation of biochemical markers does not indicate an increased risk of arterial thrombosis during use of low-dose OCs containing newer gonane progestogens; thus, the findings are in accordance with the recent epidemiological studies on these compounds. The application of relevant biochemical markers facilitate the understanding of the non-reproductive effects of sex steroids which have increasing importance because of their expanding use, not only as contraceptives, but also in the treatment of benign gynaecological disorders, as hormone replacement therapy and as prophylactic agents against specific degenerative conditions. Moreover, they may prove to be helpful in the future identification of women, who have increased susceptibility to the metabolic effects of sex steroids due to genetic predisposition. PMID- 11894724 TI - Cholesterol reduction in patients with lower limb atherosclerotic disease. AB - Patients with peripheral arterial disease (PAD) have a 2-3 times increased risk of death and in the most severe stage, critical peripheral ischaemia, the mortality rate is around 50% within 4-5. This poor survival rate is due to concomitant coronary and cerebrovascular atherosclerotic disease. Among the major risk factors for atherosclerosis are dyslipidaemia, smoking, hypertension and diabetes. Large randomised trials have shown that dyslipidaemia is easily modifiable in both patients with and without established coronary artery disease, with significant reductions in cardiovascular morbidity and mortality. Although none of these trials directly measured peripheral vascular status, there is every indication that conclusions submitted for patients with ischaemic heart disease can be translated to patients with peripheral vascular disease. The object of this review was therefore to divulge current evidence available supporting active treatment of dyslipidaemia in patients with peripheral vascular disease. PMID- 11894725 TI - Two decades of violence. A cohort study from the Danish municipality of Aarhus. AB - INTRODUCTION: Data collected from hospitals and the police has on several occasions proved to be useful in gaining knowledge about violence. The two casualty departments, Institute of Forensic Medicine and the police in Aarhus have co-operated in three earlier studies about interpersonal violence, the latest in 1993-1994. The purpose of this recent study is to reveal updated information about the incidence and character of violence in Aarhus. METHODOLOGY: Dynamic cohort study during the period the 1st of April 1999 to the 31st of March 2000. Persons who after having been exposed to violence arrived at the casualty wards and Institute of Forensic Medicine were registered and interviewed. Also data from the police was collected. All results were compared to corresponding data collected from equally conducted studies in 1981-1982, 1987-1988, and 1993 1994. RESULTS: The number of victims significantly decreased to 1496. The annual incidence rate decreased to 4.6 victims/1000. The decrease in violence was particularly among the 15-24 year old males. The extent of violence against women remained unchanged in 1999-2000. Foreigners were significantly higher represented among victims. The character and severity of violence were unchanged. CONCLUSION: Similar findings with overall decreasing incidence of violence, and on the other hand increasing violence among foreigners have been found in similar studies from Norway and England. Preventive actions should be regarded against the 15-24 year old males and foreigners. PMID- 11894726 TI - A three year population based survey of paediatric mechanical ventilation in east Denmark. AB - BACKGROUND: East Denmark has a population of 396,000 children 0-14 years and a yearly birth rate of 30,000, but at present no paediatric intensive care unit (PICU). OBJECTIVE: To perform a population based survey of paediatric mechanical ventilation with the purpose of providing the background for discussions for or against centralization of paediatric intensive care. METHODS: Case records of children 0-14 years treated with mechanical ventilation from January 1996 to December 1998 were retrospectively reviewed and the following data were obtained: Whether or not the child was settled in East Denmark, date of admission, gender, age, underlying chronic condition(s), acute condition(s) leading to mechanical ventilation, duration of positive pressure ventilation, duration of endotracheal intubation, length of stay in ICU, and outcome. Children undergoing mechanical ventilation because of neonatal problems, cardiac surgery or neurosurgery were excluded. RESULTS: Data were obtained from 197 children of which 123 were boys (p < 0.001 for boys vs girls). Median age at admission to ICU was 30 months. Boys were younger than girls (median age 22 vs 41 months, p = 0.01), but as determined by mortality, duration of positive pressure ventilation, intubation and stay in ICU there were no differences between boys and girls with respect to disease course (p > 0.28). Totally, 86 (44%) had at least one underlying chronic condition. The incidence of disease leading to mechanical ventilation in children in East Denmark was estimated to 1.6/10,000/year. An average of 1.1 child was intubated each day. Taking into account the seasonal variation two beds would be required to give coverage for 85% of ICU days needed for paediatric mechanical ventilation while three beds would cover 98%. Children admitted to referral hospital RH more often had underlying chronic conditions and had more severe courses of disease than children admitted to other hospitals (p < 0.001). Mortality did not differ (p = 0.66). CONCLUSION: The number of children requiring mechanical ventilation in East Denmark is too low to provide the background for establishing an independent PICU. However, since paediatric intensive care is a rare and complicated event further centralization of children undergoing mechanical ventilation in East Denmark should be considered. PMID- 11894727 TI - Intensive physical therapy after hip fracture. A randomised clinical trial. AB - INTRODUCTION: This randomised study evaluates the effect of intensive physical therapy on the duration of rehabilitation following hip fracture. METHODS: Eighty eight patients transferred for rehabilitation after surgical treatment for hip fracture were included in the trial. Forty-four patients were randomised to physical therapy 3.6 hours (median) a week, while the 44 control patients received physical therapy 1.9 hours a week. Outcome was defined as duration of physical rehabilitation until the patient was able to (1) walk 50 metres in less than 2 minutes, (2) manage stair climbing to the first floor, (3) manage sit-to stand transfer, (4) move in and out of bed, (5) manage bathing, dressing and lavatory visits. RESULTS: In the group randomised to intensive physical therapy 24 patients withdrew after 15 days while 13 patients withdrew from the control group after 22 days (median values). Early withdrawal was due to orthopaedic complications, general weakness and poor co-operation. No difference between the two groups was demonstrated in the duration of physical rehabilitation by a per protocol analysis of the patients who completed the trial. DISCUSSION: The considerable drop-out rate suggests that intensive physical therapy may be of limited value when attempting to reduce the duration of rehabilitation following hip fracture. An altered objective including enhanced out-patient rehabilitation may be necessary in order to reduce the length of hospital stay after hip fracture. PMID- 11894728 TI - Combination therapy in the treatment of allergic rhinitis. AB - Allergic rhinitis is a common allergic condition. There are a variety of pharmacologic treatments, including antihistamines, oral decongestants, and intranasal corticosteroids. Leukotrienes cause significant nasal obstruction. Leukotriene receptor antagonists decrease symptoms and improve quality of life in patients with seasonal allergic rhinitis. Similar to antihistamines, antileukotrienes appear to be less efficacious than nasal corticosteroids. Combination therapy of histamine and leukotriene antagonists produces symptomatic improvement as well as improved quality of life. Areas of study for combination antimediator therapy include expanding the initial findings with regard to nasal steroids, investigation of patient preference and compliance, use in perennial allergic rhinitis, and treatment of "one airway," i.e., treatment of concurrent allergic rhinitis and asthma. PMID- 11894729 TI - Intravenous gamma-globulin therapy in bronchial asthma. AB - Two possible uses exist for intravenous gamma-globulin (IVIG) therapy in asthma. First, it has been suggested that high-dose IVIG can serve as an anti inflammatory, immunomodulatory agent in steroid-dependent asthma patients. Second, IVIG can be used as a replacement treatment in those asthma patients with frank hypogammaglobulinemia or more subtle antibody deficiencies. The mechanisms by which IVIG functions are widely different in these two potential uses. Clear characterization of the patients' immune status is pivotal in choosing whether to use IVIG. The assessment should not be limited to simple determination of serum immunoglobulin A (IgA), IgG, IgM, and IgG subclass levels. When clinically warranted, the specific antibody response to active immunization with antigens such as those in Pneumovax may be invaluable in identifying patients with subtle antibody-deficiency disorders. Asthma in those patients may be improved markedly if infection is prevented by antibody-replacement therapy with IVIG. PMID- 11894730 TI - Inflammatory mediators in cystic fibrosis lung disease. AB - Cystic fibrosis (CF) is a complex multisystem disorder caused by mutations in a membrane glycoprotein called the CF transmembrane regulator (CFTR), which has as its major function serving as a Cl- channel. The relationship between defects in CFTR and development of lung disease remains incompletely understood. Chronic lung disease, characterized by persistent infection with a peculiar type of Pseudomonas aeruginosa, bronchiectasis, and airway obstruction is the major cause of morbidity and mortality in CF patients. The inflammatory response to the chronic infection resembles that induced by lipopolysaccharide (LPS) and is mediated primarily by cytokines such as tumor necrosis factor (TNF), interleukin 1 (IL-1), IL-6, and IL-8, whose synthesis is activated by the transcription factor nuclear factor kappa B (NF-kappa B). Large numbers of neutrophils dominate the inflammatory response and excessive concentrations of their products create a vicious cycle that becomes injurious rather than protective and eventually claims the life of the patient. PMID- 11894731 TI - Psychogenic cough in adults: a report of two cases and review of the literature. AB - Psychogenic cough, also known as "habit cough," is a well-documented condition in the pediatric and adolescent population, with numerous cases reported in the medical literature. Many of these patients are strikingly similar in their clinical characteristics and, although the data are limited, a variety of treatment options may be successful in terminating this form of cough. However, psychogenic cough in adults has been reported infrequently and is less well defined. We report two cases of psychogenic cough in adult patients referred to our service for an evaluation of refractory, chronic cough and review the relevant medical literature. Our patients seemingly represent the first cases of psychogenic cough reported in the geriatric population and share clinical features with children, adolescents, and young adults. One case is unique in the sense that the cough responded to a distracter in the form of a throat lozenge, and this patient consumed > or = 20 lozenges/day for approximately 13 years. Psychogenic cough should be considered in adult patients who present with a chronic cough of no obvious organic basis that has failed therapy directed at postnasal drip, asthma, and gastroesophageal reflux. We propose criteria to assist in making a diagnosis of psychogenic cough in adult patients and review the limited information that exists concerning treatment modalities. PMID- 11894732 TI - Prevalence of allergen-specific IgE among patients with chronic fatigue syndrome. AB - The prevalence of atopy among patients having chronic fatigue syndrome (CFS) has been reported to be as high as 80% in published surveys of patients with this syndrome. However, many of the reports relied on self-assessment by patients for the presence of atopy or solely used total immunoglobulin E (IgE) levels to assess the likelihood of atopy. To more critically assess the presence of atopy among patients with CFS, testing was done for total IgE and allergen-specific IgE using the Pharmacia CAP system including 20 common allergens: trees (birch/oak/ash), grass (rye/blue), weeds (common/giant ragweed), molds (Penicillium/Aspergillus/Alternaria), dust mites (Dermatophagoides pteronyssinus/Dermatophagoides farinae), animal dander (cat/dog), and foods (egg white/milk/wheat/corn/peanut/shrimp). Testing of 50 patients having documented CFS indicated that 78% had total IgE < 100 IU/mL, among whom 26% had a positive test for allergen-specific IgE of class I or greater for one or more allergens. Among the 22% of CFS patients having a total IgE > 100 IU/mL, 73% had a positive test for allergen-specific IgE for one or more allergens. The most commonly positive allergens were dust mites (24-26%), whereas molds (0-6%) and foods (0 4%) were rarely positive. The overall frequency of positive results for the presence of allergen-specific IgE among CFS patients was 36%, not significantly different from the normal prevalence of these antibodies in the general population (20-35%). This assessment of the prevalence of allergen-specific IgE antibodies in patients with CFS fails to support a potential association between CFS and atopy. PMID- 11894733 TI - A novel case of mealworm-induced occupational rhinitis in a school teacher. AB - A 47-year-old African American female elementary schoolteacher presented with itchy, watery eyes, rhinorrhea, postnasal drainage, and nasal congestion complicated by recurrent epistaxis for 2 months. She had similar symptoms the previous year from September to May but was symptom free during the summer. Her symptoms began within 1 hour after entering the classroom and improved in the evening at home, on weekends, and vacation. She denied symptoms around dust, freshly cut grass, or pets and had no prior history of underlying allergic rhinitis and asthma. She had a 20-pack-a-year smoking history but quit 1 1/2 years ago. A detailed history of her classroom environment revealed the presence of mealworms that were used to teach the children about life cycles. Physical exam revealed swollen, erythematous nasal turbinates but was otherwise unremarkable. Prick skin testing was positive for oak tree, grasses, feathers, and cockroaches. Mealworm whole body extracts were prepared using standard methodology. Titration intracutaneous skin testing revealed a positive reaction at a 1:1000 concentration associated with a large delayed reaction 8 hours later that persisted for 24 hours. Specific nasal provocation using acoustic rhinometry revealed a dose response change in nasal volume (48% decrease at 1:100; 53% decrease at 1:50) and cross-sectional area (32% decrease at 1:100; 48% decrease at 1:50) in response to mealworm challenge compared with a saline control. Removal of the mealworms from the classroom resulted in complete relief of her symptoms. This is the first reported case of mealworm-induced rhinitis in a schoolteacher. Because mealworm demonstrations are now part of the standard curriculum in public school elementary classrooms in Ohio, it is important that school administrators recognize the sensitizing nature of these insects and their potential for causing allergic rhinitis and asthma in the workplace. PMID- 11894734 TI - Azelastine's inhibition of histamine and tryptase release from human umbilical cord blood-derived cultured mast cells as well as rat skin mast cell-induced vascular permeability: comparison with olopatadine. AB - Mast cells are involved in early and late-phase reactions by releasing vasoactive molecules, proteases, and cytokines. Azelastine and olopatadine are histamine 1 receptor (H-1R) antagonists with antiallergic effects present in the ophthalmic solutions Optivar and Patanol, respectively. Because it is difficult to obtain animal or human conjunctival tissue, we first investigated the effect of these compounds on histamine and tryptase release from cultured human mast cells (CHMCs) grown out of human umbilical cord blood-derived CD34+ cells. Sensitized CHMCs were pretreated with various concentrations of azelastine or olopatadine for 5 minutes. Then, CHMCs were challenged with anti-immunoglobulin E (IgE) and the released mediators were quantitated. The greatest inhibition of mediator release was seen when CHMCs were pretreated with 24 microM of azelastine or 133 microM of olopatadine (2% dilution of azelastine or 5% olopatadine original ophthalmic solutions, respectively). We then studied the drug concentrations that gave optimal results on skin vasodilation induced by the mast cell secretagogue compound 48/80. An intradermal injection of 48/80 in rats, to which Evan's blue had been administered via the tail vein, induced substantial dye extravasation. Pretreatment of the injection site for 5 minutes with either 24 microM of azelastine or 133 microM of olopatadine completely prevented extravasation; this effect was quantitated also by fluorometric assessment of Evan's blue extracted in formamide. Evaluation of skin mast cells from injected sites showed that mast cell degranulation was inhibited greatly. These results indicate that on an equimolar basis, azelastine was a more potent inhibitor than olopatadine of both CHMC and rat skin mast cells activation. PMID- 11894735 TI - Recombinant allergens for immunotherapy. AB - Many of the problems associated with using natural allergenic products for allergy diagnosis and treatment can be overcome using genetically engineered recombinant allergens. Over the past 10 years, the most important allergens from mites, pollens, animal dander, insects, and foods have been cloned, sequenced, and expressed. Allergens have diverse biological functions (they may be enzymes, enzyme inhibitors, lipocalins, or structural proteins). High-level expression systems have been developed to produce recombinant allergens in bacteria, yeast, or insect cells. Recombinant allergens show comparable immunoglobulin E (IgE) antibody binding to natural allergens and show excellent reactivity on skin testing and in in vitro diagnostic tests. Recombinant allergens will enable innovative new strategies for allergen immunotherapy to be developed. These include peptide-based vaccines, engineered hypoallergens with reduced reactivity for IgE antibodies, nucleotide-conjugated vaccines that promote Th1 responses, and the possibility of developing prophylactic allergen vaccines. PMID- 11894736 TI - Common variable immunodeficiency. AB - We present a case report and review of the literature that illustrates many key features of patients with common variable immunodeficiency (CVID). These patients frequently present with repeated infections with a variety of different microorganisms. Recurrent sinopulmonary infections can lead to serious chronic complications such as bronchiectasis, and gastrointestinal infections can result in malabsorption. In addition to serious infection, CVID is associated with a number of comorbid disorders including a variety of autoimmune diseases and neoplasms. Here, we provide an illustrative case report and discuss the primary features and therapy for patients with CVID. PMID- 11894737 TI - Straws in the wind latin writings on asthma between Galen and Cardano. PMID- 11894738 TI - Effects of air pollutants on the allergic response. AB - There is strong evidence that the prevalence and severity of allergy are increasing in many areas of the world. Pollutants have been implicated as one of the important factors responsible for these observations because they have been shown to act at multiple steps in the allergic response. They can increase immunoglobulin E (IgE) synthesis and potentiate the specific IgE response to antigens. Pollutants also are capable of inducing a primary specific IgE response to an antigen and increase allergic airway inflammation. In addition, pollutants can induce airway inflammation or cause nonspecific irritation. Pollutants also are capable of modulating the allergenicity of airborne substances, which possibly may promote allergic sensitization. PMID- 11894739 TI - Mediterranean diet: the past and the present. AB - Mortality statistics from the World Health Organization have provided early evidence that diet in the Mediterranean countries has been affecting the health of the respective populations and, in particular, their coronary health. Keys (1) has taken the lead arguing that the traditional Mediterranean diet has beneficial effects on health. Recent studies, capturing the evidence accumulated over the last three decades, have documented that the traditional Mediterranean diet meets several important criteria for a healthy diet. An attempt to conceptualize the proper diet and to make it able to function has been reported and a score has been developed and evaluated. Studies among the elderly in Greece, Denmark, Australia, Spain and China have shown that the overall Mediterranean dietary pattern was more important for longevity than single nutrients. These findings suggest, therefore, that a Mediterranean diet is associated with longer survival. Two additional questions should be addressed at this time: Is the Mediterranean diet an integral entity, or the sum of identifiable components that can and should be separately considered in the development of guidelines? Is the Mediterranean diet or its major components transferable to populations living far from the Mediterranean area? Answers to these questions would be important for scientific and policy reasons. The dietary patterns that prevail in the Mediterranean area have many common characteristics, most of which stem from the fact that olive oil plays an important role in all of them. Thus, although different regions in the Mediterranean basin have their own diets, it is legitimate to consider them as variants of a single entity, the Mediterranean diet. The Mediterranean diet can be described as the dietary pattern found in the olive growing areas of the Mediterranean region, in the late '50s and early '60s, when the consequences of World War II were overcome, but the fast-food culture had not reached the area yet (2). Olive oil is important both because of its several beneficial properties and because it allows the consumption of large quantities of vegetables and legumes in the form of salads and of cooked foods. Other essential components of the Mediterranean diet are wheat, grapes, and their derived products. Total lipid consumption may be high, around 40% of total energy intake as in Greece, or moderate, around 30% of total energy intake as in Italy. In all instances, however, the ratio of monounsaturated to saturated dietary lipids is much higher than in other places of the world, including northern Europe and North America (3). PMID- 11894740 TI - Nutrition and health: epidemiology of diet, cancer and cardiovascular disease in Italy. AB - Most epidemiological data suggest a protective role for fruits and vegetables in the prevention of several common epithelial cancers, including digestive and major non-digestive neoplasms. The relation between frequency of consumption of vegetables and fruit and cancer and myocardial infarction risk was analysed using data from a series of case-control studies conducted in Italy. For digestive tract cancer, population attributable risks for low intake of vegetables and fruit ranged between 15 and 40%. A selected number of antioxidants showed a significant inverse relation with breast and colorectal cancer risk, although the main components responsible for the favourable effect of a diet rich in vegetables and fruit remain undefined. Fish tends to be another favourable indicator of reduced cancer risk. In contrast, subjects reporting frequent red meat intake showed a relative risk consistently above unity for several common neoplasms. Whole grain food intake was consistently related to reduced risk of several types of cancer, particularly of the upper digestive tract neoplasms. Epidemiological evidence of the relation between fiber and colorectal cancer indicated a possible protections. In contrast, refined grain intake was associated to increased risk of different types of cancer, pointing to a potential role of insulin-like growth factor 1 (IGF-1). A low risk diet for cardiovascular disease includes high consumption of fish, vegetables and fruit, and hence rich in ascorbic acid and other antioxidants, thus sharing several aspects with a favourable diet for cancer. PMID- 11894741 TI - The BRAVO project: screening for childhood obesity in a primary school setting. AB - BACKGROUND AND AIM: The prevalence of childhood obesity is increasing worldwide, raising a number of public health concerns. First, childhood obesity is a strong predictor of adult obesity; second, the low long term success rate and the high social cost of the treatment of obesity suggest that attention should be paid to the prevention of obesity early in childhood. The objective of the present study was to evaluate dietary habits and anthropometric factors in a sample of schoolchildren aged 6-12 years living in Southern Italy in the framework of an ongoing prospective study aimed at childhood obesity prevention. METHODS AND RESULTS: The BRAVO Project is carried out in co-operation with the school staff and is a part of an educational program on nutrition for the schoolchildren and their families. During the first phase of the study, the prevalence of overweight and obesity was evaluated in 363 children adopting the criteria for definition of childhood obesity recently proposed by the International Obesity Task Force (IOTF). The main result of this study was that in our cohort an exceedingly high risk of becoming overweight in adult age was observed for any one-year age class. CONCLUSIONS: The results of the present study confirmed the trend toward an increasing prevalence of childhood obesity observed in other Western countries. Moreover, out data--though preliminary--suggest that the large scale involvement of primary school in screening programs could represent an effective preventive strategy against the increased risk of childhood obesity. PMID- 11894742 TI - The "Food, GI-tract functionality and human health" European Research Cluster, PROEUHEALTH. PMID- 11894743 TI - Dietary and genetic influences on susceptibility or resistance to weight gain on a high fat diet. AB - This paper outlines the concepts and objectives behind the Fifth Framework Programme 'Diet and Obesity', scheduled to run from February 2001 to January 2004 with financial support from the European Commission. The objective is to identify the causes of weight gain on an energy-dense, high fat diet, and apply findings in the diagnosis, prevention and management of obesity. Molecular, physiological, behavioural and clinical approaches will be applied to the study of the mechanisms underlying diet preferences and susceptibility to weight gain, in both humans and laboratory rodents. This strategy is based on the observation that not all individuals habitually eating a high fat diet are obese; some have a similar body mass index (BMI) to low fat consumers despite the consumption of substantially more fat (and energy). The project will investigate why some individuals preferentially select a high fat diet, and how genetic background interacts with diet to confer susceptibility or resistance to obesity. Improved diagnosis of individuals at risk may allow treatment and preventive measures, including advice during pregnancy, to be more targeted. The effect of early life nutrition on dietary preferences, susceptibility to obesity, and the programming of brain signalling systems will be investigated in parallel clinical and rodent studies. PMID- 11894744 TI - Study of the regulation by nutrients of the expression of genes involved in lipogenesis and obesity in humans and animals. AB - Dietary digestible carbohydrates are able to modulate lipogenesis, by modifying the expression of genes coding for key lipogenic enzymes, like fatty acid synthase. The overall objective of the Nutrigene project (FAIR-CT97-3011) was to study the efficiency of various carbohydrates to modulate the lipogenic capacity and relevant gene expression in rat and human species (control and obese subjects) and to understand the underlying molecular mechanisms involved in the regulation of lipogenic genes by carbohydrates. Key cellular mediators (namely SREBP-1c and 2, AMP activated protein kinase, cholesterol content) of the regulation of lipogenic gene expression by glucose and/or insulin were identified and constitute new putative targets in the development of plurimetabolic syndrome associated with obesity. In humans, hepatic lipogenesis and triglyceride synthesis, assessed in vivo by the use of stable isotopes, was promoted by a high carbohydrate diet in non obese subjects, and in non alcoholic steatotic patients, but was not modified in the adipose tissue of obese subjects. Non digestible/fermentable carbohydrates, such as fructans, were shown to decrease hepatic lipogenesis in non obese rats, and to lessen hepatic steatosis and body weight in obese Zucker rats. If confirmed in obese humans, this would allow the development of functional food able to counteract the metabolic disturbances linked to obesity. PMID- 11894745 TI - Dietary habit profile in European communities with different risk of myocardial infarction: the impact of migration as a model of gene-environment interaction. The IMMIDIET Study. AB - The risk of myocardial infarction (MI) is lower in southern than in northern European countries. The lower rate of MI in the Mediterranean regions of Europe suggested a potential role of the traditional Mediterranean diet in the prevention of MI. Unfortunately, in the last 20 years, a tendency to adopt Westernised food habits even in southern regions of Europe is reflected by an increase in the prevalence of obesity. Therefore the impact of diet on MI risk profile among European populations needs to be reconsidered. Genetic risk factors have also been implicated in the development of MI. Genes, indeed, continuously interact with environmental factors in determining the pathogenesis of MI. The aims of the IMMIDIET study are to evaluate: 1. The present dietary habits and the risk profile of three European communities at different risk of MI; 2. The impact of migration on risk factors for MI. Dietary habits and genetic polymorphisms will be evaluated in an Italian, Belgian and British population sample. The historical Italian migration to Belgium and the integration through mixed marriage will be considered as a model of gene-environment interaction. As an index of MI risk profile, factors that are most likely under the combined influence of both dietary and genetic determinants will be investigated. PMID- 11894746 TI - Perspectives for food research and European collaboration in the European Research Area and the new Framework Programme. AB - Since 1987, successive framework programmes have contributed to strengthen European food research through the establishment of networks between research institutions, universities and companies from various European countries. In the FAIR programme (1994-1998), 118 research projects comprising nearly 1,000 participants from the European Union and Associated States have been supported in the food area with a European funding of about [symbol: see text] 108 million. Within the Quality of Life and Management of Living Resources programme (1998 2002), food research is mostly supported within the key action 'food, nutrition and health' with a budget of [symbol: see text] 290 million. After the first four deadlines, 735 eligible research proposals have already been received. Further to their evaluation by a panel of independent experts, 108 proposals have been funded or selected for funding representing a total contribution of about [symbol: see text] 168 million. Among those, several clusters of projects are now running on important topics such as probiotics, coeliac diseases, mycotoxins, GMO, safety and food for the elderly. In addition, technology stimulation measures are largely benefiting SMEs to foster their innovation potential. In January 2000, the European Commission adopted a Communication entitled "Towards the European Research Area (ERA)" with the objective to contribute to developing better framework conditions for research in Europe. On 21 February 2001, the Commission adopted proposals to be submitted to the European Parliament and Council for the next framework programme for research and innovation (2002-2006). The new framework programme that is becoming one of the financial instruments of the ERA aims at catalysing the integration of European research by: strengthening of links between the Community research effort and national and regional research policies; concentrating on a limited number of priority fields or research to which activities at the Union level can add real value. One of the seven priority areas, entitled 'food safety and health risks', is intended to help establish the integrated scientific and technological bases needed to develop a system of production and distribution of safe and healthy food and control food-related risks, relying in particular on biotechnology tools, as well as health risks associated with environmental changes. A total budget of [symbol: see text] 600 million is proposed for this priority. In the priority areas, the new framework programme will work mainly by supporting the development of cooperation within networks of excellence bringing together the best research capabilities in Europe's regions to conduct common research programmes and integrated projects involving public and private partners, with clearly stated scientific and technological objectives. PMID- 11894747 TI - Functional Food Science in Europe. AB - The goal of the Functional Food Science in Europe (FUFOSE) concerted action was to reach consensus on scientific concepts of functional foods in Europe by using the science base that supports evidence that specific nutrients positively affect physiological functions. The outcome proposes "a working definition" of functional foods: foods can be regarded as functional if they can be satisfactorily demonstrated to affect beneficially one or more target functions in the body, beyond adequate nutritional effects, in a way relevant to an improved state of health and well-being and/or reduction of risk of disease. Functional foods must remain foods and they must achieve their effects in amounts normally consumed in a diet. Evidence from human studies, based on markers relating to biological response or on intermediate endpoint markers of disease, could provide a sound scientific basis for messages and claims about the functional food products. Two types of claims are proposed that relate directly to these two categories of markers: Enhanced function claims (type A) and reduced risk of disease claims (type B). A new EU Concerted Action will start with, and build upon, the principles defined within FUFOSE. This project PASSCLAIM will (i) produce a consensus on principles for the scientific substantiation of health related claims for food and food components, (ii) select common criteria for how markers should be identified, validated and used in well-designed studies to explore the links between diet and health and (iii) to evaluate critically the existing schemes which assess the scientific substantiation of claims. PMID- 11894748 TI - Measurements of consumer attitudes and their influence on food choice and acceptability (AIR-CAT). AB - A changing European food market demands insight into consumer attitudes and their influence on food choice and acceptability. This multidisciplinary area needs to bring together scientists from all regions of Europe and with very different scientific backgrounds. The primary objectives of this concerted action have been: to establish a base with state of the art methods for measurements of consumer attitudes; to review and test existing methods in practical applications in collaboration with European food industries; to perform comparative studies between laboratories on food products, where attitudes play different roles for consumer behaviour in the community countries, such as transgenic foods, irradiated foods, foods with different additives, declarations and process technologies, foods with different origin declarations, ecological foods and foods with strong health connotations (such as high-fat foods). The members of the action have published more than 130 publications related to aspects of how consumer attitudes can be measured and how food choice behaviour is related to acceptability, during the last four years. Studies have been conducted in relation to methodological aspects as well as particular studies related to specific food items and regions for food production. The paper will give a brief selection of relevant results from experiments reported through the action. During 2001 a textbook called "Food, People and Society, in a European Perspective", will be published. The book was initiated during the action and is later supported with additional authors. Altogether 29 chapters will cover the whole spectrum of topics from consumer food choice and acceptability to market perspectives and risk analysis. PMID- 11894749 TI - HealthSense: how changes in sensory physiology, sensory psychology and sociocognitive factors influence food choice. AB - HealthSense is a multi-centre shared cost research project in sensory and consumer sciences. This project was initiated in response to changing population demographics, which predict a significant increase in the older age group. The primary objective of HealthSense is to determine accurately the effects of ageing on the perceptual abilities of the sensory systems, and ultimately how age related change determines sensory preferences and food choice. The project is carried out by 24 participating institutes located in 10 different European countries. The project is funded by the European Commission Quality of Life Fifth Framework Programme (QLKI-CT-1999-00010). PMID- 11894750 TI - Compatibility of household budget and individual nutrition surveys: results of the preliminary analysis. AB - The EU-supported project entitled: "Compatibility of household budget and individual nutrition surveys and disparities in food habits" aimed at comparing individualised household budget survey (HBS) data with food consumption values derived from individual nutrition surveys (INS). The present paper provides a brief description of the methodology applied for rendering the datasets at a comparable level. Results of the preliminary evaluation of their compatibility are also presented. A non parametric modelling approach was used for the individualisation (age and gender-specific) of the food data collected at household level, in the context of the national HBSs and the bootstrap technique was used for the derivation of 95% confidence intervals. For each food group, INS and HBS-derived mean values were calculated for twenty-four research units, jointly defined by country (four countries involved), gender (male, female) and age (younger, middle-aged and older). Pearson correlation coefficients were calculated. The results of this preliminary analysis show that there is considerable scope in the nutritional information derived from HBSs. Additional and more sophisticated work is however required, putting particular emphasis on addressing limitations present in both surveys and on deriving reliable individual consumption point and interval estimates, on the basis of HBS data. PMID- 11894751 TI - The FAIR-INNOVATION dissemination project. AB - The goal of the FAIR-INNOVATION dissemination project (FLAIR-FLOW 3) (1997-2000) was to disseminate R&D results from the EU FAIR programme to small and medium sized food enterprises (SMEs), health professionals (HPs) and consumer groups (CGs) in 19 European countries. The dissemination routes were: (i) one-page technical documents on research results; (ii) their reproduction on the internet; (iii) their reproduction in journals Europe-wide; (iv) workshops on results from EU-supported food research programmes; and (v) lectures and poster presentations by FLAIR-FLOW network personnel. Of the 135 one-pagers produced, 62% were targeted at SMEs, 18% at HPs and 5% at CGs. The remaining 15% were on food safety and were common to the three target groups. There were 1047 publications arising from one-pages in trade journals, and over 8000 requests (paper route) were received for follow-up information. These were in addition to 240 k downloads from the FLAIR-FLOW 3 web site (www.flair-flow.com). Initiatives for HPs included specially collated versions of the one-pagers for major conferences, 20 focused workshops, and interaction with the European Federation of Associations of Dieticians. Currently, dissemination is continuing through FLAIR-FLOW 4 (2001 2003) in 24 countries. FLAIR-FLOW 4 is co-ordinated by M. Jean Francois Quillien from INRA-CRIAA (FR) at criaa@rennes.inra.fr. PMID- 11894752 TI - On assessment of success and the IMPACT of FAIR. AB - Early in 2001, the authors were awarded an Accompanying Measure by the Agri-Food Directorate of DG Research to assess the IMPACT of a selected subset of projects recently completed within the FAIR RTD program. We have three aims: To specifically evaluate the impact of 25 projects supported under the EU FAIR Programme. To develop and validate a methodology for assessing the impact of collaborative projects in the food area. To identify any generic messages which could relate to the forthcoming European Research Area (ERA). A series of Workshops are being held with FAIR participants and stakeholders around the Community to facilitate the development of the methodology. At the time of writing meetings have been held in the UK, Ireland and Denmark. Diet and Health will a key driver for innovation in food during the 21st century and will play a major role in both EU and national research. Participation in the HEALFO Food and Health Conference allowed us to meet and discuss IMPACT with an important group of both past and future contributors to EU research. This paper outlines the workshop methodology and outlines some interim findings together with emerging hypotheses. PMID- 11894753 TI - Wine and cardiovascular disease. AB - The objective of the European project FAIR CT 97 3261 "Wine & Cardiovascular Disease" was to have insight into the biological effects and mechanisms of wine constituents as protective factors against cardiovascular disease and to evaluate their clinical relevance in the context of a regular and moderate wine consumption (1, 2). The project aimed at evaluating the effects of red wine polyphenolic extracts (RWPE) on the major components contributing to the pathophysiology of atherosclerosis and thrombosis, namely vascular tone, haemostatic system, oxidative processes, and plaque development, in selected in vitro, ex vivo, and in vivo models (3-6). Evidence was obtained that RWPE might improve vascular function mainly through nitric oxide (NO)-mediated mechanisms, interfere with haemostatic and oxidative processes involved in the progression of vascular damage, and modulate early events of atherosclerosis. These effects were studied on cellular and plasmatic blood components and on vascular function in experimental animal models. A study in 40 healthy volunteers, performed in Barcelona, showed a significant increase in HDL cholesterol (HDL-C) levels and a decreased oxidation of LDL after red wine ingestion (30 g alcohol daily for 4 weeks) as compared to the same amount of alcohol given as spirit. A significant fall was seen, after red wine, in the expression of several markers of cardiovascular risk measured on circulating cell surface or in plasma of volunteers. Finally, a meta-analysis of all available epidemiological studies indicated a significant negative association of moderate wine consumption (150 300 ml daily) with the risk of major cardiovascular events. In conclusion, this FAIR project contributed to the establishment of a biological plausibility to the epidemiological association of moderate wine consumption with prevention of cardiovascular disease. Red wine and RWPE altogether may prove to be beneficial against the major killer of European citizens. PMID- 11894754 TI - Foods for tomorrow: the challenge for nutrition. AB - Diet related diseases, including cardiovascular disease, cancer, osteoporosis, and degenerative changes of ageing are a major cause of morbidity and mortality throughout Europe. There is considerable potential to improve public health by means of diet. This includes changing eating behaviour at the population level, through the development and implementation of nutrition policy and promulgation of dietary guidelines for healthy eating. This must be complemented by measures which modify the nature of the food supply. Europe's food industry has a central role in facilitating dietary change by providing health promoting foods, including novel and functional foods, which are safe, convenient, palatable and acceptable to consumers. The development of policies, programmes and products for improving public health will need to be informed by innovative research which strengthens our fundamental understanding of the relationships between food and physiological functions related to health, as well as of factors which influence food choice and eating behaviour. This is a considerable challenge for nutrition and will require the organisation of research to provide for the integration of appropriate disciplines and expertise, including recent advances achieved in molecular and cell biology and in the analysis of the human genome. PMID- 11894755 TI - Beneficial nutritional properties of olive oil: implications for postprandial lipoproteins and factor VII. AB - Previous research concerning protective cardiovascular properties of olive oil has focussed on the beneficial consequences on blood cholesterol levels of substituting dietary saturated fatty acids with oleic acid. Despite evidence implicating raised circulating triglycerides in the postprandial state in the pathogenesis of atherosclerosis and thrombosis, little research had been conducted to investigate effects of monounsaturated fatty acids on postprandial events. In a case control study of southern (n = 30) versus northern European (n = 30) men, significant differences in postprandial triglyceride and apolipoprotein (apo) B-48 response were observed, with evidence of attenuated and potentially beneficial responses in the Southern Europeans. In a randomised controlled study manufactured foods typical of the Northern European food culture, were used to deliver diets rich in either saturated or monounsaturated fatty acids (from olive oil). During the period of the olive oil enriched diet, LDL-cholesterol levels were 15% lower (p < 0.001) than during the saturated fat diet. Postprandial triglyceride response was shifted towards the profile seen in southern European men and the postprandial activation of factor VII, as well as the production of factor VII antigen, was reduced on the olive oil diet. The study demonstrated significant improvements in biomarkers for cardiovascular disease in subjects osed to high olive oil diets (Southern Europeans) or transferred to such diets in the short term (Northern European volunteers). The study produced novel findings with respect to potential mechanisms by which diets high in monounsaturated fatty acids (MUFA) can reduce population risk of cardiovascular disease. PMID- 11894756 TI - Garlic and health. AB - Garlic is a worldwide cultivated crop species which is well-known for its health beneficial effects. In this paper an overview is given on a number of important plant and health aspects of garlic. Furthermore an outline is presented of a European initiative which is aimed at the development of high quality garlic and the identification of biomarkers in atherosclerosis and cancer in humans for disease prevention. PMID- 11894757 TI - The relative importance of traditional and "modern" foods for Israeli Negev Bedouins. A population in transition. AB - The impact of urbanization on the health and nutritional status of developing populations is an issue of concern worldwide. The Bedouin Arabs in Southern Israel are a traditionally semi-nomadic/nomadic population undergoing a rapid process of urbanization which is accompanied by rising chronic disease rates. We examined the diet of urban and rural (more traditional) Bedouins to determine the relative importance of modern foods and drinks in their daily diet. We found that for main meals both populations rely heavily on traditional foods (available before 1948) but for snacks and drinks many manufactured products are used. These products rich in calories may contribute to the changing disease patterns. Rural areas rely more on traditional milk products which are made non-perishable, while urban populations use more meat products and pre-prepared meals because of the availability of electricity and refrigeration. Traditional foods and drink and the traditional way of eating (eating from a common plate) is still a very important part of the Bedouin way of life. This pattern of eating requires the development of culture-specific dietary assessment methodology to allow quantitative assessment of both traditional and more modern foods consumption and handle the eating practices of the Bedouin society today. PMID- 11894758 TI - Vitamin A, vitamin E and carotenoid status and metabolism during ageing: functional and nutritional consequences (VITAGE PROJECT). AB - Among the nutritional factors contributing to maintain health during ageing, fat soluble vitamins (FSV) are crucial to protect against free radical-generated degenerative processes or impaired efficiency of the immune system. However, no sound scientific evidence is able to confirm specific dietary needs in vitamin A, vitamin E and carotenoids for the healthy elderly. VITAGE project aims at providing such evidence by undertaking studies on male volunteers from 3 European countries, aged between 20-75 years. Biomarkers and variables related to status, metabolism and functions will be measured either in steady-state conditions, or during dietary depletion and repletion in FSV. Original, yet already developed, methodologies will provide clear information about the physiological characteristics of vitamin A, vitamin E and carotenoids. Simultaneously, marketing opportunities for FSV-enriched dietetic foods, specifically designed for the elderly will be determined. The scientific and economical evidence obtained in this project will provide the basis to implement a EU nutritional policy towards the elderly and to develop a new sector of dietetic food products. PMID- 11894759 TI - Towards a strategy for optimal vitamin D fortification (OPTIFORD). AB - The research project "Towards a Strategy for Optimal Vitamin D Fortification (OPTIFORD)" has received financial support from the 5th framework programme of the EC. Vitamin D deficiency is a serious health problem among large population groups in Europe. Subjects segregated from sun exposure for social or religious reasons, or because of disability are at risk for vitamin D deficiency. Vitamin D is essential for proper bone mineralisation and vitamin D deficiency is recognised to be an important risk factor for hip fracture. Fortification and/or supplementation strategies could be effective and inexpensive ways of arresting preventable health consequences. However, fortification policies in Europe differ and this reflects the many unknowns in relation to the strategy of vitamin D fortification of food, particularly concerning the levels achieving optimal effects without toxicity. The overall objective of OPTIFORD is to investigate if fortification of food with vitamin D is a feasible strategy to remedy the insufficient vitamin D status of large population groups in Europe, and to determine at what level fortification should be pitched. An important outcome is to reinforce the scientific basis for recommendations on vitamin D as a nutrient. PMID- 11894760 TI - Increased fruit and vegetable consumption: potential health benefits. AB - In this European Union project, a Core human study was conducted in Ireland, Northern Ireland, Spain, France and The Netherlands. Oxidative and antioxidant status, vegetable and fruit consumption, and carotenoid intake of volunteers from different countries were compared. Response to increased carotenoid intake was determined. Attention was paid to whether the antioxidant capability of beta carotene, lutein and lycopene demonstrated in vitro was apparent in relation to increased oxidation resistance of low-density-lipoprotein (LDL). Other (complementary) studies were undertaken and included determination of: protective effects of carotenoid-rich foods against LDL and DNA oxidative damage; carotenoid absorbability; barriers to increased vegetable consumption; and carotenoid content of fruits and vegetables frequently consumed in Europe. Our results demonstrated that carotenoid supplementation did not increase LDL oxidation resistance. However, increased consumption of carotenoid-rich fruits and vegetables did increase LDL oxidation resistance, and higher plasma concentration of total and specific carotenoids (pre-supplementation) was associated with lower DNA damage. PMID- 11894761 TI - Heterocyclic amines: human carcinogens in cooked food? AB - During the frying of meat and fish, genotoxic heterocyclic amines (HCAs) are formed. The dietary exposure to HCAs may be implicated in the aetiology of human cancer, but there may be other factors in our diet that prevent the genotoxic effects of these compounds. Within the project described here, we plan to identify regional and individual cooking habits that affect HCA-levels in our food. These are determined with a validated analytical method and the exposure to HCAs is estimated by dietary assessment. Biomarker analysis will be employed to estimate recent or long-term exposure to HCAs. In order to identify genetically determined risk factors in humans, cell lines are genetically engineered expressing allelic variants of acetyl- and sulfotransferases implicated in HCA metabolism. Species differences of metabolism and toxicity of HCAs are assessed and the influence of the intestinal microflora on HCA-induced toxicity is evaluated. Dietary constituents that may reduce the genotoxicity of HCAs are screened for potential protective effects in in vitro and in vivo model systems. Finally, we will aim at human intervention studies to investigate if these protective factors are relevant for man. The objectives of this project are to estimate and possibly reduce the exposure levels to HCAs in Europe, to identify populations highly susceptible to HCA toxicity, and to reduce the toxic effects of HCAs by protective factors. PMID- 11894762 TI - Evaluation in human volunteers of the potential anticarcinogenic activities of novel nutritional concepts: prebiotics, probiotics and synbiotics (the SYNCAN project QLK1-1999-00346). AB - Prebiotics is a recent novel food concept that includes food ingredients that are not digested in the human upper intestinal tract and hence arrive in the colon where they are selectively fermented by a limited number of colonic bacteria. Amongst these are bifidobacteria and lactobacilli, which are considered indicators of a well-balanced intestinal flora. Probiotics are bacteria that, while passing through the intestine, may exert specific beneficial effects on the host's physiology. In general, probiotics are members of the group of the lactic acid-producing bacteria. By means of a variety of experimental models it was demonstrated that prebiotic carbohydrates and probiotics consistently reduced processes of carcinogenesis and tumorigenesis. Synergistic chemopreventive actions were observed with combinations of the two, which together are called synbiotics. One of the most important causes of death in the ageing western population is colon cancer, which is typically associated with a western-style diet. On the basis of the available experimental data, an EU-funded research project (the SYNCAN project QLK1-1999-00346) was set up to evaluate whether synbiotics and prebiotics can be added to food without detriment to (and hopefully eventually improving) organoleptic properties. They are, as such, a good vector for importing nutritionally interesting properties into our diet. PMID- 11894763 TI - Functional food ingredients against colorectal cancer. An example project integrating functional genomics, nutrition and health. AB - Functional Food Ingredients Against Colorectal Cancer is one of the first European Union funded Research Projects at the cross-road of functional genomics [comprising transcriptomics, the measurement of the expression of all messengers RNA (mRNAs) and proteomics, the measurement of expression/state of all proteins], nutrition and human health. The goal of Functional Food Ingredients Against Colorectal Cancer is to develop a colon epithelial cell line-based screening assay for nutrients with presumed anti-colorectal carcinogenic properties. Genes involved in colon carcinogenesis are identified at the RNA and protein level, using a variety of methods (subtractive hybridisation, DNA microarray, proteomics) in combination with models for colorectal cancer development (human biopsies, rat model for colorectal carcinogenesis, colorectal cancer epithelial cell lines). Secondly, colorectal cancer epithelial cell lines are selected, in terms of their capacity to undergo gene/protein expression changes representing different phases in the colorectal carcinogenesis. Thirdly, these cell lines are used to determine the effects of nutrients with presumed anti-carcinogenic properties (e.g. resveratrol, flavonoids) on functional genomics-derived endpoints. Once validated against the effects of these nutrients in in vivo animal models and classical biomarkers for colorectal carcinogenesis, these cell line models combined with functional genomics represent useful tools to study colorectal carcinogenesis and screen for nutrients with anti-carcinogenic properties. PMID- 11894764 TI - Early malnutrition and programming of adult degenerative diseases: experimental, epidemiological and preventive studies. PMID- 11894765 TI - Pharyngoesophageal pressure in patients with swallowing disorders. AB - We developed and tested a pressure transducer to correctly determine swallowing pressure at different sites in the pharynx. In normal individuals three pressure peaks were obtained in the mesopharynx, hypopharynx and cervical esophagus, respectively. A patient with central dysphagia demonstrated markedly low mesopharyngeal and hypopharyngeal swallowing pressure. One tongue cancer patient who had undergone right hemiglossectomy, including partial resection of the root of the tongue and bilateral superior neck dissection, had markedly low swallowing pressure in the mesopharynx and vallecula area. Another supraglottic cancer patient treated by supraglottic horizontal partial laryngectomy showed extremely low swallowing pressure in the supraglottic area. Based on our findings, we suggest that measurements using a pressure transducer such as the one described here should be used in combination with radiographic study to diagnose swallowing anomalies correctly. Data obtained with the pressure transducer will allow the clinician to identify the site responsible for postoperative dysphagia as well as its severity, and facilitate planning of reconstructive surgery when required. PMID- 11894766 TI - Factors that affect swallowing-related apnea times in humans. AB - The dynamics of deglutition were studied in relation to potential changes due to aging. Swallowing-related apnea time (SAT) was measured during "dry" and "wet" swallowing in 84 adults without dysphagia to examine if age-related variation of SAT corresponded to changes in deglutition dynamics due to aging and to determine possible significance. Swallowing movements were recorded using a transducer for measuring swallowing pressures. Respiratory flow rates during deglutition were measured with a heat-wave flowmeter as part of phonatory function testing system. Respiratory movements were recorded with a respiration pick-up band set at the diaphragm. Findings demonstrated that SAT did not change with aging but was prolonged in subjects over age 50 years. When topical anesthesia was omitted during study, SAT tended to be shorter during wet swallowing than during dry swallowing in all age groups. Statistically significant differences were observed between wet and dry swallowing SAT in subjects under the age of 39 years. When a topical anesthetic was applied to pharyngeal mucous membranes, SAT was prolonged. Finally, SAT results during wet swallowing determined in a anesthetized dog before and after topical anesthesia of the pharyngeal mucous membranes showed significant prolongation after topical anesthesia. These findings suggest that the sensation of the pharyngeal mucous membrane may be important in producing swallowing-related apnea and controlling its duration. PMID- 11894767 TI - Low-dose intratympanic gentamicin treatment of Meniere's disease. AB - To control attacks of vertigo while preserving both hearing and labyrinthine function, low doses of gentamicin were instilled intratympanically in nine patients with intractable unilateral Meniere's disease. Each patient received six instillations of antibiotic of 4 mg each (total dose, 24 mg). Patients were then followed for 2-4 years. Long-term results of treatment are reported according to the American Academy of Otolaryngology-Head and Neck Surgery 1985 criteria. Of the nine cases, three experienced complete control of vertiginous attacks, while six received substantial control. Post-treatment hearing acuity was unaffected, although disability following treatment became worse in one patient, a 66-year old man. Caloric responses after therapy were absent or severely reduced in three ears, moderately reduced in two ears and unchanged in four ears. In three patients, labyrinthine function was found damaged 4-8 days after administration of the last dose of drug. Overall, intratympanic instillations of low doses of gentamicin in patients with intractable Meniere's disease were found to control vertiginous attacks with less damage to the inner ear function than that reported in the literature. PMID- 11894768 TI - Neural projections from the frontal cortex to the oculomotor nucleus: an anatomical study using retrograde axonal, anterograde axonal and transneuronal transport of wheat germ agglutinin-conjugated horseradish peroxidase in cats. AB - Neural projections from the frontal cerebral cortex to the oculomotor nucleus (3N) were investigated in 1- to 2-year-old cats by retrograde and anterograde axonal and transneuronal transport of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Following injection of WGA-HRP into the 3N area and its surrounding tissues, retrogradely labeled cells were observed in the anterior sigmoid gyrus, ventral bank of the cruciate sulcus, medial and lateral walls and base of the presylvian sulcus, gyrus rectus and gyrus proreus. Following injection of WGA-HRP into these frontal cortical areas, anterogradely labeled nerve terminals were observed in the mesencephalic periaqueductal gray matter (PAG) just overlying the 3N. Only a few terminals were observed within the 3N. Following injection of WGA-HRP into the extraocular muscles of 1-month-old kittens, transneuronally labeled small cells were observed in the PAG just overlying the 3N and in the mesencephalic reticular formation, ventrolateral to it. These small cells may represent intercalated neurons of the cortico oculomotor projections in the cat. PMID- 11894769 TI - Effects of stimulation of the vestibular nuclei on posterior hypothalamic neuron activity in guinea pigs. AB - To clarify the differences among the four main vestibular nuclei in the vestibulo autonomic reflex, we examined the effects of electrical stimulation of superior, lateral, medial and descending vestibular nuclei (SVN, LVN, MVN and DVN) on posterior hypothalamic area (PHA) neurons in the guinea pig. Ipsi- and contralateral SVN stimulation produced excitation in 30% and 25% of the PHA neurons tested, respectively. Twenty percent of the PHA neurons showed an excitatory response to ipsilateral LVN stimulation while 60% of the neurons tested responded to contralateral LVN stimulation, including excitation of 36% and inhibition of 24%. MVN and DVN stimulation produced little change in PHA neuron activity. These findings suggest that vestibular information processed in the SVN and the LVN is conveyed to the hypothalamus and may then contribute to activation of the vestibulo-autonomic reflex. PMID- 11894771 TI - Pupillary responses in normal subjects following auditory stimulation. AB - To clarify pupillary responses of humans following auditory stimuli, we studied both eyes of 61 normal subjects using a computed pupillograph. Unilateral auditory stimulation elicited pupillary dilatation in all cases. Pupillary responses were classified according to duration as being either "long" or "short". The duration of dilatation was 1530 +/- 320 ms (mean +/- SD) in the long lasting group (n = 45) and 850 +/- 250 ms in the short-lasting group (n = 16). The latency time for dilatation was 460 +/- 80 ms. Both eyes of each subject showed the same response. Two drops of 10% guanethidine, a sympathetic blocking agent, were applied to one eye of 3 subjects. Although the early phase of dilatation was barely affected, the late phase was inhibited, as seen in long lasting dilatation. The short-lasting response was unaffected. We conclude that the long-lasting response consists of an early pupillary dilatation due to inhibition of parasympathetic nervous activity and a late dilatation due to excitation of sympathetic activity. The short-lasting response is produced only by inhibition of the parasympathetic component. PMID- 11894770 TI - A 16-year survey of changes in bacterial isolates associated with chronic suppurative otitis media. AB - Bacteria were isolated and cultured from the ears of outpatients with chronic suppurative otitis media (OMC) who attended the otologic clinics of Kyushu University Hospital from 1976 to 1991. Bacterial isolates were analyzed retrospectively to determine their incidences and antibiotic sensitivities. Staphylococcus aureus and Pseudomonas aeruginosa were the most commonly isolated organisms. The incidence of S. aureus infections gradually rose over 16 years, while the incidence of Proteus infections gradually declined. The frequency of isolation of glucose non-fermenting gram-negative rod (NF-GNR) species, fungi and anaerobic bacteria gradually increased. The species of bacteria isolated from OMC cases with acute and chronic infections, cholesteatoma and postoperative infections were compared. Over the 16 years, the isolation of S. aureus increased in frequency. The differences in each type of OMC ceased to exist. S. aureus were quite sensitive to most of the antibiotics tested, whereas Ps. aeruginosa and NF GNR showed poor sensitivity. PMID- 11894772 TI - Ultrastructure of guinea pig stria vascularis processed by rapid freezing and freeze substitution. AB - The rapid-freezing and freeze-substitution method fixes a specimen as if it were prepared before excision. We used this method to study the stria vascularis of guinea pigs using electron microscopy. Findings were essentially the same as those obtained with conventional chemical fixation, although freeze substitution made it possible to observe the membrane structures in a smoother and more linear manner. This method seems to be the procedure of choice for studying the instantaneous movement and behavior of cells. PMID- 11894773 TI - Fine structure of the human cochlear aqueduct: a light and transmission electron microscopic study of decalcified temporal bones. AB - The morphologic features of the human cochlear aqueduct were examined using both light and electron microscopy. The lumen of the cochlear aqueduct was observed to be filled with dense, irregular connective tissue corresponding to dura mater. At the entrance to the cerebrospinal fluid space, the dense connective tissue in the ductal lumen was covered with a thin layer of a few flattened cells, which was contiguous with the arachnoid membrane of the brain. A simple low cuboidal epithelium also separated the perilymphatic space from the lumen of the duct. Our observations confirm the presence of a barrier membrane at the opening to the perilymphatic space, and suggest that no transport occurs in the human cochlear aqueduct. PMID- 11894774 TI - Profiles of resting potentials across the stria vascularis in kanamycin-deafened guinea pigs. AB - The resting potentials of the marginal cells in the stria vascularis of the guinea pig were determined from changes in the combined electrode-tissue resistance of the electrode. The resistance of the electrode was 45.5 +/- 16.0 M omega (n = 20) before penetration of the stria vascularis and 46.7 +/- 17.3 M omega (n = 20) after penetration. The resistance drops across the luminal membrane of the marginal cells were 46.0 +/- 22.6 M omega (n = 12) in kanamycin deafened guinea pigs and 54.5 +/- 33.1 M omega (n = 9) in normal guinea pigs. The endocochlear potential (EP) and resting potentials in the marginal cells were 90.1 +/- 6.0 mV (n = 14) and 70.4 +/- 11.3 mV (n = 14) in kanamycin-deafened guinea pigs and 84.8 + 5.1 mV (n = 29) and 74.7 +/- 11.7 mV (n = 29) in normal guinea pigs. The resting potentials in the marginal cells decreased gradually and were approximately 0 mV around 20 min after anoxia in both kanamycin-deafened and normal guinea pigs. These changes were comparable to those of EP in kanamycin deafened guinea pigs during anoxia. The mechanism of the EP in kanamycin-deafened guinea pigs is discussed. PMID- 11894775 TI - Potassium circulation in the perilymph of guinea pig cochlea. AB - To determine whether any differences exist in potassium circulation between the scala vestibuli and scala tympani, we recorded the change in K+ activity in both scalae of the guinea pig cochlea at the basal and third turns, using a double barrelled, K(+)-sensitive microelectrode after perfusion with artificial perilymph containing 20 mM KCl and 130 mM NaCl. K+ activity increased immediately after the start of perfusion and decreased after its completion. The rates of decrease of K+ activities were approximately 1.0 mEq/l per min in the scala vestibuli of the basal and third turns, also 1.0 mEq/l per min in the scala tympani of the basal turn, and approximately 0.5 mEq/l per min in the scala tympani of the third turn. The rate of decrease of K+ activity in the scala tympani was significantly slower in the third turn than in the basal turn. Blockage of the cochlear aqueduct depressed the rate of decrease of K+ activity in the scala tympani more in the basal turn than in the third turn. These results suggest that there is a difference in potassium circulation between the scala vestibuli and scala tympani, and that the cochlear aqueduct plays an important role in potassium circulation in the perilymph of the scala tympani. PMID- 11894776 TI - The effects of ototoxic drugs on mechano-electric transduction channels in chick cochlear hair cells. AB - The effects of ototoxic drugs on mechano-electrical transduction (MET) currents were investigated in dissociated cochlear hair cells of the chick, using a whole cell patch-electrode voltage-clamp technique. Dihydrostreptomycin (DHSM) and cisplatin (cis-dichlorodiammine platinum II, CDDP) blocked the MET channel in a dose- and voltage-dependent manner. In contrast, acetyl salicylate did not suppress the MET current. At -50 mV, DHSM and CDDP blocked the MET channel with a Hill coefficient of 0.93 and 2.1, respectively. These findings suggest that a single DHSM molecule or more than one CDDP molecule binds to a single MET channel. PMID- 11894777 TI - Voltage-dependent channels in dissociated outer hair cells of the guinea pig. AB - Voltage-dependent channels in outer hair cells (OHCs) dissociated from the guinea pig cochlea were investigated by the use of a whole-cell patch-clamp technique. Two types of K+ current were recorded from OHCs. One was a slowly inactivating K+ current that was activated at a potential more positive than -30 mV. Another is a K+ current that was already activated at resting membrane potential. After suppressing both K+ currents, depolarizing voltage steps elicited a slowly inactivating inward current that was dependent on external Ca2+ and was indicative of an L-type Ca2+ channel in OHCs. Aminoglycoside antibiotics known to be ototoxic selectively inhibited the Ca2+ current. PMID- 11894778 TI - Effects of acoustic overstimulation on cochlear evoked potentials. AB - Guinea pigs were exposed to 2 kHz pure-tone or octave-band pass noise at an intensity of 100 dBSPL for 30 min. The effects of sound exposure on cochlear microphonics (CM) and compound action potential (AP) were studied using a test condition devised to complete the measurement of the sensitivity of both potentials for the frequency from 1 to 7 kHz within several minutes. The loss of CM sensitivity was limited to around 5 dB for all test frequencies in animals exposed either to pure-tone or band noise. In contrast, the loss of AP in both exposure conditions was significantly greater than that of the CM, and the magnitude of the AP losses reflected the frequency characteristics of the exposure sounds. From these observations, the AP is considered to be a more sufficient index than the CM in studying the effects of acoustic overstimulation. PMID- 11894779 TI - Topical applications of trichloroacetic acid as therapy for nasal allergy. AB - A group of 22 patients (27 nostrils) with nasal allergies was treated with 80 w/v trichloroacetic acid applied to the inferior turbinates. Patients were then evaluated prospectively based on both subjective responses and clinical examinations. Allergic symptoms were reduced significantly, especially those involving nasal obstruction and watery rhinorrhea. Nasal airflow resistance also improved after treatment (P < 0.001). Nasal provocation testing revealed a significant decrease in post-treatment responses (P < 0.001). No severe side effects were noted after treatment. Findings demonstrated that local application of trichloroacetic acid is a safe, effective and simple treatment for outpatients with symptomatic nasal allergies. PMID- 11894781 TI - Comparison of vestibulo-ocular reflexes in earth-horizontal and earth-vertical axis rotations. AB - Ten normal subjects and nine patients with peripheral labyrinthine lesions were subjected to sinusoidal rotation about an earth-horizontal (EHA) and earth vertical axis (EVA) using a new device. In normal subjects the gain of the vestibulo-ocular reflex (VOR) in EHA rotation was significantly larger than that of the VOR in EVA rotation. This difference may be attributed to the interaction of the semicircular canals and otolithic organs. Unlike normal subjects, patients with unilateral lesions showed no differences in gain between EHA and EVA rotations toward either the intact or abnormal side. Patients with bilateral lesions also showed no differences in gain. These observations suggest that the interaction of the semicircular canals with the otolithic organs enhancing VOR gain does not occur if one of the otolithic organs is defective in either ear. This does not appear to recover once such a dysfunction develops. PMID- 11894782 TI - Use of a remote controlled radiotherapy afterloading system to manage unresectable, metastatic thyroid cancer in the trachea. AB - We treated a 66-year-old Japanese woman with metastasis from thyroid papillary adenocarcinoma to mediastinal lymph nodes that obstructed the airway by its extensions towards the tracheal carina. Since the patient suffered from severe dyspnea, an endotracheal intubation was inserted and endotracheal radiotherapy was done using a remote controlled afterloading system. The obstruction mass in the trachea almost disappeared and there has been no evidence of regrowth during a follow-up period fo more than 3 years. The patient's management is now detailed. PMID- 11894780 TI - Segregation analysis of IgE responses to Cryptomeria japonica pollen antigen in vivo. AB - The IgE response to Cryptomeria japonica pollen antigen (CPAg) in vivo was determined by radioimmunoassay of the plasma of 525 members from 98 families with known nasal allergies. Based on responses, patients were classified into a non responder or low-responder group (non/low) and a high-responder group. Segregation analysis revealed that the IgE non/low responsiveness to CPAg involved a single dominant trait. The gene frequency was calculated to be 0.44 0.60. The IgE non/low response to CPAg was found to be mediated by CPAg-specific suppressor T cells. These findings demonstrated that the phenotypic variation of IgE responsiveness to CPAg is not due the immune response gene, but rather is mediated by the immune suppression gene for CPAg, via CPAg-specific suppressor T cells. PMID- 11894783 TI - Argyrophilic nucleolar organizer regions as a prognostic indicator of laryngeal carcinomas. AB - Nucleolar organizer regions (NORs) were studied in surgically removed specimens from 76 patients with squamous cell carcinomas of the larynx using an argyrophilic (Ag) staining technique. The mean number of AgNORs per nucleus was 4.3 +/- 1.38 (SD). The mean AgNOR number for T1 or T2 disease was statistically lower than that for T3 or T4 lesions (P < 0.05). The mean AgNOR number was lower in patients with N0 disease than in patients with N1 or N2 or N3 tumors (P < 0.05). There was also a statistically significant difference between the mean AgNOR number for stage II and stage III disease (P < 0.01), for stage III and stage IV disease (P < 0.05), but not for stage I and stage II disease. According to the histological grading, there was a significant difference between the mean AgNOR number for the well-differentiated and moderately differentiated tumors (P < 0.05), and for the moderately differentiated and poorly differentiated types (P < 0.01). These results suggest that the number of AgNORs in the nucleus is a significant indicator of laryngeal carcinomas. PMID- 11894784 TI - A survey of the pattern of glandular distribution in the larynges of human infants. AB - We studied the development of glandular structures and the pattern of distribution of serous and mucous-type glands in the larynges of 77 human infants obtained at autopsy. The subglottic larynx was subjected to quantitative and qualitative analysis, using an image binarization procedure. The areas of mucosa and inner space of the subglottic larynx tended to increase with age. The area occupied by the glands differed considerably in younger infants and the areas of total glands and serous-type glands increased with age. There was no correlation between the area of the mucous glands and age. These findings indicate that local immune function of the secretory glandular system in the larynx of human infants increases with advancing age. PMID- 11894785 TI - Preliminary findings of chromosomal alterations and expression of cell cycle genes in head an neck tumors. AB - The genesis and progression of malignant tumors may be related to certain somatic mutations and the accumulation of multiple chromosomal alterations. Using four freshly resected malignant tumors, we investigated the relationship between chromosomal alteration and expression of cell cycle regulatory genes. Specimens of thyroid hyperplasia and normal thyroid tissue were also investigated. As cell cycle regulating genes, we chose the cdc2 gene that encodes the p34cdc2 protein kinase, a major kinase of the cell cycle, and the RCC1 gene that is essential for coupling between S and M phases. Three of the malignant tumors contained cells with chromosomal alterations, including one polyploid and two aneuploid. The DNA content of cells in thyroid hyperplasia was the same as in the normal gland. The amount of p34cdc2 protein was very low in cells of both normal thyroid and hyperplastic tissue, and grew very slowly as compared with malignant tumors. There was no significant relationship between the amount of RCC1 and ploidy pattern. PMID- 11894786 TI - Carotid body tumor associated with differentiated thyroid carcinoma. AB - We report a case of a carotid body tumor associated with a primary differentiated thyroid carcinoma. A 44-year-old woman presented with a 10-year history of an asymptomatic mass in her neck. Physical examination revealed a pulsatile submandibular mass in her right neck as well as multiple nodules in the thyroid. Magnetic resonance imaging, computed tomography and, in particular, angiography were diagnostic of the carotid body tumor. Slight changes in serum thyroglobulin levels and thyroid scintigraphy led us to suspect thyroid carcinoma. Embolization of the arteries feeding the carotid body tumor was performed, and was followed by tumor resection 24 h later. At surgery, histopathology confirmed the presence of follicular and papillary carcinomas of the thyroid, resulting in concurrent resection of the gland. There were no residual cranial nerve deficits. The patient subsequently received radiotherapy. Diagnosis and surgical management are discussed, together with pathogenetic factors. PMID- 11894787 TI - Immunocytochemical studies of major gap junction proteins in rat salivary glands. AB - We examined protein components of the gap junctions between acinar cells of the parotid, sublingual and submandibular glands of the rat, using type-specific antibodies directed against major gap junction proteins, connexin32 (Cx32) and connexin26 (Cx26). Double-immunofluorescence microscopy revealed that fluorescent spots of both connexins in the parotid and sublingual glands were distributed between the apposed regions of acinar cells. They appeared together, or were co localized. The intensity of the Cx26-associated fluorescent signals was relatively weak in the submandibular glands compared with the other glands and was absent from some acini. When present, these spots were always co-localized with Cx32 immunoreactive positive spots. The results suggest that Cx32 and Cx26 in rat salivary glands are colocalized within the same gap junctional plaques when simultaneously expressed by the same acinar cells. PMID- 11894788 TI - Preventing fractures in large rural centres: sociodemographic sub-groups at risk of osteoporosis from their lifestyle. AB - Middle-aged people living in non-metropolitan Victoria have higher hospitalisation rates from osteoporotic fractures than those in metropolitan areas. This may reflect a higher prevalence of lifestyle risk factors for osteoporosis. One-fifth of Victoria's non-metropolitan population live in 'large rural centres'. The aim of the present study was to identify the sociodemographic sub-groups in a 'large rural centre' at risk of osteoporosis because of their lifestyle. Adults on Ballarat's electoral rolls were invited to complete a questionnaire and have their height and weight measured. A total of 335 eligible people participated in the present study (67% response). The sub-groups at risk of osteoporosis were identified using logistic regression analyses. Among women, being single/separated/divorced/widowed was associated with being underweight and having low dietary calcium. A lack of exercise was associated with not completing high school and smoking with being aged 25-44 years. Among men, low dietary calcium was associated with not completing high school and smoking was associated with being employed in a non-professional/non-managerial occupation. These sub groups must be considered when planning preventative strategies for people in 'large rural centres'. PMID- 11894789 TI - Evaluation of a pilot project to use computers in a rural general practice term. AB - In 1998, the Department of General Practice (Faculty of Medicine and Dentistry, University of Western Australia) ran a pilot project to use computers in a sixth year rural general practice term. Students were provided with a laptop computer to take into rural and remote areas throughout Western Australia during their 4 week clinical attachment. An email mailing list was set up for course participants to share experiences and complete set learning activities specifically related to rural general practice. An evaluation of this pilot project found that students felt less isolated on rural attachments, course outcomes were improved and rural preceptors were more involved in the programme. The development of a teaching and learning programme that involves the use of computers in rural general practice for undergraduate students has the potential to improve the quality of their medical education. PMID- 11894790 TI - Differences in cardiac procedures among patients in metropolitan and non metropolitan hospitals in New South Wales after acute myocardial infarction and angina. AB - An observational cohort study examined the difference in use of cardiac procedures during and after hospital admission for acute chest pain in 47 metropolitan or non-metropolitan hospitals across New South Wales (NSW). There were 3836 patients, represented by 4151 admissions to hospital after acute myocardial infarction (AMI), unstable angina or other angina. Follow up at 22 months was completed on 1695 patients. Patients admitted to metropolitan hospitals had higher rates of most cardiac procedures while in-patients than did patients in non-metropolitan hospitals. Odds ratios (95% confidence intervals) for the use of exercise stress tests, echocardiograms, nuclear studies and coronary angiography were 3.30 (1.38, 7.90), 9.34 (4.07, 21.44), 4.87 (2.08, 11.39) and 68.64 (17.29, 272.49), respectively, for patients with AMI and 1.93 (0.91, 4.12), 5.60 (1.60, 19.57), 3.51 (1.48, 8.33) and 38.57 (9.36, 158.94), respectively, for patients with unstable angina. Rates were similar between hospital types during the 22 months after discharge. The appropriateness of this large variation in resource use between metropolitan and non-metropolitan hospitals requires examination. PMID- 11894791 TI - Meningococcal infections and meningitis: what is new? AB - Meningococcal infection is one of the very few severe bacterial infections, in this era, that still can kill a relatively healthy child within minutes. Fortunately, it is a relatively rare disease. Rural practitioners may see one affected child once every 2-3 years, but once seen they will never forget it. The present article gives some examples of case scenarios along with a brief overview of the problem, with emphasis on early diagnosis, prevention and possible future developments. PMID- 11894792 TI - Learning issues for nurses in renal satellite centres. AB - The introduction of 'satellite' dialysis centres has increased local access to renal services for patients living in rural and remote areas across Australia and is, therefore, consistent with rural health policies. Rural health strategies also aim to maintain and improve the skills of health professionals through regular review of the scope of their practice and evaluation of the education required. Yet, the results of the present national study of nurses working in satellite dialysis centres indicate that, for many, the context of practice influences their ability to access ongoing professional education to support and extend their nursing practice. The present study showed that there was a range of educational backgrounds among nurses working in rural 'satellite' centres with reportedly limited access to specific dialysis-related professional development. In the present paper we report on factors that influence the ability of nurses working in satellite dialysis centres to access ongoing professional education. PMID- 11894793 TI - Selected demographic, social and work characteristics of the Australian general medical practitioner workforce: comparing capital cities with regional areas. AB - The aim of the present study was to compare selected characteristics of the Australian general medical practitioner workforce in capital cities and regional areas. Data were derived from the 1996 Census of Population and Housing. Characteristics included age, sex, full- or part-time work, place of birth and change in residential address. Analyses were performed for each state and territory in Australia, the statistical division containing each capital city and all other statistical divisions in each state and territory. Of the 26,359 general medical practitioners identified, 68% were male. More female than male general medical practitioners were aged < 45 years (74 vs 52%, respectively; P < 0.0001). The proportion of general medical practitioners aged < 35 years was higher in capital cities (30%) than regional areas (24%; P < 0.0001). Overall, 32% of the general medical practitioner workforce was female and almost 50% of those aged < 35 years were female. The proportion of female general medical practitioners was higher in capital cities than regional areas, by up to 30%. While 13% of male general medical practitioners reported part-time work, 42% of females also reported part-time work and these figures were similar in capital cities and regional areas. Approximately 40% of the Australian general medical practitioner workforce was born outside Australia and while fewer migrants have entered in recent years they were more likely to be living in regional areas than the capitals. The census provides useful medical workforce data. The regional workforce tends to be made up of more males and is older than in capital cities. Monitoring trends in these characteristics could help to evaluate initiatives aimed at addressing regional workforce issues. PMID- 11894794 TI - The hidden power of the rural nursing profession. PMID- 11894795 TI - Toward an increased understanding of user demographics in online sexual activities. AB - A large study (N = 7,037) was conducted through the MSNBC website during June 2000 to explore the general profile of people who engage in online sexual activity (OSA) as well as gender differences in OSA. The study explored ways in which the Internet is used for sexual activities, reasons for engaging in OSA, and consequences of use. The study found a relationship between use of OSA and its consequences in users' lives as well as significant gender differences in user profiles. Significant gender differences were found in reasons respondents gave for engaging in OSA and OSA-related behaviors. Overall, these findings increase our understanding of the user profiles of people who engage in OSA and corroborate gender differences in offline research. Implications for further research are discussed. PMID- 11894797 TI - Assessing the role of relationship conflict in sexual dysfunction. AB - Relationship conflict has long been thought to cause, maintain, and influence the therapeutic outcome of sexual problems in the absence of a physical cause. The results of conflict can influence partners' relationship satisfaction, and relationship satisfaction can influence sexual satisfaction. General relationship deficiencies, such as unresolved conflict, undermine the mutual acceptance that is important to healthy sexual functioning. The purpose of this article is to summarize some of the basic empirical findings of studies of conflict patterns in relationships and their role in sex dysfunction and to suggest a model for assessing relationship conflict as a feature of sexual dysfunction. Results from several studies indicate that couples with sexual problems may have conflict management issues and employ distinct conflict-resolution styles compared to satisfied couples. Dysfunctional conflict resolution may be a cause or result of some sexual problems, whereas constructive interaction concerning conflict can add to emotional and sexual intimacy in a couple's relationship. These patterns warrant systematic attention in assessment and intervention in sex therapy. PMID- 11894796 TI - Bupropion-sustained release as a treatment for SSRI-induced sexual side effects. AB - Twenty-four subjects treated with serotonin reuptake inhibitors for a depressive disorder who had new-onset sexual side effects coincident with antidepressant treatment were treated with escalating doses of bupropion SR up to 300 mg daily for 7 weeks. Global response rates were 46% for women and 75% for men. All sexual side effects improved in response to bupropion SR in both men and women with no differential effect on any one sexual side effect. Most of the improvement (more than 50%) occurred within the first 2 weeks and at low dose (100-200 mg/day). When prescribed in an open fashion, bupropion SR appeared to be effective in treating all the major categories of sexual side effects. PMID- 11894798 TI - Split gender identity: problem or solution? Proposed parameters for addressing the gender dysphoric patient. AB - Working with the gender dysphoric patient is complex because of the various clinical issues that arise. One issue that has not been addressed in the psychiatric literature is whether to address the patient with the biologically congruent pronoun or name or with the patient's preferred-gender pronoun or cross gender name. This article presents clinical examples that allow a template to be developed for pronoun use in working with such patients. Whether the clinician uses biologically congruent names and pronouns may depend upon the patient's progress in adopting the cross gender role as well whether family or friends either know or accept such changes. In certain situations, such as meetings with family members, the therapist may address the patient with gender congruent names; whereas on other occasions use cross-gender pronouns or names. PMID- 11894799 TI - Erectile dysfunction and depression: category or dimension? AB - Depression, as a risk factor for erectile dysfunction (ED), has received minimal systematic attention. One-hundred twenty men with ED evaluated in a sexual behaviors clinic were studied. The categorical Diagnostic and Statistical Manual of Mental Disorders (DSM-IV; American Psychiatric Association, 1994) diagnosis of a depressive disorder was found in only 14 subjects (14.7%). Dimensional quantification of depression was measured with the Brief Symptom Inventory (BSI). The BSI data revealed clinically significant elevations of depression and other dysphoric affects. The presence of a comorbid medical diagnosis did not affect the rates of categorical diagnosis of depression or the dimensional levels. The five factors of personality in the NEO-PI were within normal range. The data demonstrates that men with ED are affectively distressed but infrequently meet criteria for categorical DSM-IV depression. PMID- 11894803 TI - Abstracts of the Physiological Society scientific meeting held at University of Bristol, 5-7 September 2001. PMID- 11894802 TI - Abstracts of MEDNET 2001, the 6th World Congress on the Internet in Medicine. Udine, Italy, 29 November-2 December 2001. PMID- 11894804 TI - Science communication--a process of mutual education. PMID- 11894800 TI - Evaluating changes in sexual functioning in depressed patients: sensitivity to change of the CSFQ. AB - Accurately evaluating alterations in sexual functioning requires a validated instrument that measures clinically relevant change over time. One-hundred one depressed patients from 15 Spanish out-patient clinics completed the Changes in Sexual Functioning Questionnaire (CFSQ; Clayton, McGarvey, & Clavet, 1997) at baseline and after 6 months of treatment with fluoxetine, nefazodone, paroxetine, or venlafaxine. Sexual desire/interest showed a nearly substantial floor effect (30% of patients indicated the maximum score) for women in the nefazodone group at baseline and in the paroxetine group at final visit. The percentage of dimensions recording change was greater for women (80%) than for men (20%) in the nefazodone group (improving changes) and greater for men (40%) than for women (20%) in the paroxetine group (worsening changes). Highest effect sizes were found on sexual desire/frequency with improvement in women in the nefazodone group (SES = 0.49), and on orgasm/ejaculation with worsening in men in the paroxetine group (SES = -1.45). In conclusion, the CSFQ is sensitive to bidirectional changes and is appropriate for measuring sexual dysfunction. PMID- 11894805 TI - Gene assessment and sample classification for gene expression data using a genetic algorithm/k-nearest neighbor method. PMID- 11894806 TI - [Relations of human papillomavirus and lesions of the oral mucosa]. AB - Recently there has been an increased interest in the possible role that viruses and especially the Human Papilloma virus (HPV) could play in the etiology of lesions of the oral mucosa. A distinction has to be made between the so-called low-risk types of the virus (HPV-2, 6, 11, 13 and 32) which can be found in benign oral mucosal lesions, and the high-risk type (HPV-16), which predominantly is found in malignant oral mucosal lesions. PMID- 11894808 TI - [Mouth rinses and tooth pastes]. PMID- 11894807 TI - [The role of lactic acid bacteria in nutrition and health]. AB - Lactic acid bacteria possess both positive as well as negative aspects in relation to health. On the one side metabolic activities such as the production of extracellular polysaccharides or lactic acid from carbohydrates, are responsible for plaque formation and demineralisation of enamel. On the other hand, further down in the digestive tract, these groups of micro-organisms can have a beneficial effect by locally influencing the microbial composition, specific metabolic activities or stimulation of the immune response. Lactic acid bacteria also play a major role in food preservation. When plants or animals become food after harvesting or slaughter, endogenous resistance mechanisms such as the immune systems are lost and consequently foods become susceptible to spoilage. Food spoilage can be retarded or prevented by application of certain external factors; for example, storage at low temperatures of fermentation by lactic acid bacteria. Similarly, the mature dentition also consist of non-living tissue and is therefore also subject to 'spoilage' (decay). Lactic acid bacteria now play a negative role. However, the metabolism of lactic acid bacteria in plaque and consequently de- and remineralisation may also be influenced by application of external factors as diet, oral hygiene and fluorides. This paper will discuss the functional similarities and differences of lactic acid bacteria in different areas of the digestive tract. PMID- 11894810 TI - Nutrition and metabolism. PMID- 11894811 TI - Proceedings of the 7th annual conference of the Australian Electrophoresis Society. Sydney, Australia, November 9-10, 2000. PMID- 11894813 TI - 2001 Membership directory. PMID- 11894814 TI - [Exacerbated dyspnea]. PMID- 11894812 TI - HIV-associated lipodystrophy, metabolic complications, and antiretroviral toxicities. PMID- 11894815 TI - [A feverish soldier]. PMID- 11894816 TI - Bibliography. Current world literature. Clinical nephrology. PMID- 11894817 TI - Bibliography. Current world literature. Pathophysiology of hypertension. PMID- 11894818 TI - The 2001 Thomas Hunt Morgan Medal. Yasuji Oshima. PMID- 11894819 TI - The 2001 Genetics Society of America Medal. H. Robert Horvitz. PMID- 11894820 TI - The 2001 George W. Beadle Medal. Gerald R. Fink. PMID- 11894821 TI - [Deep ocean water]. PMID- 11894823 TI - XII European meeting of the French Society of Cardiology. 16-19 January 2002, Paris, France. Abstracts. PMID- 11894824 TI - Abstracts of the 51st annual meeting of the American College of Cardiology. Atlanta, Georgia, USA. March 17-20, 2002. PMID- 11894825 TI - Abstracts of the 3rd International Conference on Kidney and Hypertension. Madrid, Spain, 7-9 February 2002. PMID- 11894826 TI - 2001 Ostomy Wound Management Buyers Guide. PMID- 11894827 TI - Legal aspects of research with human pluripotent stem cells in Germany. PMID- 11894828 TI - Xenotransplantation in the light of animal ethics. PMID- 11894829 TI - Xenotransplantation: informed consent/contract and patient surveillance. PMID- 11894831 TI - Society for Gynecologic Investigation membership list. PMID- 11894830 TI - Umbilical cord blood stem cell transplantation -- ethical problems. PMID- 11894832 TI - Society for Gynecologic Investigation 49th annual meeting. March 20-23, 2002. Los Angeles, California, USA. Abstracts. PMID- 11894834 TI - 53rd Meeting of the German Society for Hygiene and Microbiology. 30 September-4 October 2001. Abstracts. PMID- 11894833 TI - 2001 index issue. Cumulative indexes for volumes 280-289. PMID- 11894835 TI - [Abstracts of the ALFEDIAM meeting. Strasbourg, 2002]. PMID- 11894836 TI - Isolated autoimmune response: definition, analysis of the prevalence in an outpatient rheumatology clinic, relationship to pre-clinical autoimmune disease and infections by hepatotropic viruses. AB - OBJECTIVE: To define the clinical significance of non-organ specific autoantibody positivity in patients in whom routine clinical and laboratory examinations did not detect any disease that might have caused the serological finding. METHODS: Out of 1,120 patients consecutively admitted to an outpatient rheumatology clinic, 28 were referred for the evaluation of an autoantibody positivity unrelated to the clinical status. These patients and 28 sex- and age-matched controls underwent a specific work-up with the aim of detecting any underlying infection or autoimmune disease. RESULTS: Eight of the 28 patients (28.5%) were found to be affected by a previously undetected disease: 3 chronic hepatitis C, 3 Sjogren's syndrome, and 2 autoimmune thyroiditis. The remaining 20 did not show any autoimmune or hepatic disease, although 4 of them showed active infection by HBV (n = 1) or HGV (n = 3) and 15 had had a previous infection by hepatotropic viruses (HBV, CMV or EBV). After a follow-up lasting 6-54 months, none of the last 20 patients developed any autoimmune or chronic hepatic disease. CONCLUSIONS: A diagnostic work-up is necessary in patients presenting with unexpected autoantibody positivity in order to detect an underlying pre-clinical autoimmune disease and/or unexpected hepatic infection. Patients in whom such a work-up fails to point out any condition should be further followed in order to make an early diagnosis of autoimmune disease. PMID- 11894837 TI - Insufficiency fractures in patients with chronic inflammatory joint diseases. AB - OBJECTIVE: To describe the typical sites of stress fractures in the lower extremities and pelvis in rheumatoid patients (rheumatoid arthritis, juvenile chronic arthritis, psoriatic arthritis, ankylosing spondylitis). METHODS: Thirty three patients with 52 stress fractures [mean age 44 years (range 11-73)] were studied at the authors' institution when they were being treated for their rheumatic diseases. Fourteen patients had RA, 9 JCA, 5 PsoA, and 5 SPA. Stress fractures were detected from patient documents and from series radiographs in suspected cases. In some cases magnetic resonance imaging was also performed. RESULTS: One patient presented with 5 fractures, 2 patients with 4 and 3 fractures, and 7 patients with 2 fractures each. Other patients (n = 19) had only one fracture each. The metatarsal (MT) bones were the most common site of involvement. Twenty-five of the 52 fractures were located on MT I-V. The second and third most common sites were thefibula (n = 13) and tibia (n = 6). All fractures of the lower tibia or fibula were associated with valgus malalignment of the ankle. CONCLUSION: If a patient with rheumatic disease experiences sudden and unexplained pain localised in the forefoot, above the ankle, below the knee, or in the pelvis, a stress fracture should be suspected. Patients with severe osteoporosis, high-load corticosteroid or methotrexate therapy, or marked joint deformity are at high risk of developing stress fracture. PMID- 11894841 TI - Negative life events of anxiety disordered children: bad fortune, vulnerability, or reporter bias? AB - This article seeks to examine the nature of negative life events of anxiety disordered children: to what extent are they unique, to what extent are they shared with siblings, and when they are shared, is the impact similar or different? Twenty-five anxiety-disordered children aged 8 to 13 years, referred to a child psychiatric clinic, were compared with matched non-clinical controls, and with their nearest in age nonreferred sibling aged 6 to 13 years on the number of parent-reported stressful life events. Anxiety-disordered children differ significantly from well controls in the number of negative life events reported by their parents over their lifetime, and the year preceding referral. Anxiety disordered children also differ significantly from their non-referred nearest in age sibling in the number of negative life events, both non-shared and shared. The difference in shared events is due to differences in appraisal by the parents of the impact of a shared event on the respective children. The often reported finding that children with anxiety disorders have experienced more negative life events than their healthy peers is partially due to objective differences in the occurrence of these events, but may also reflect heightened vulnerability or reporter bias. PMID- 11894838 TI - Intradermal recombinant hepatitis B vaccine for healthcare workers who fail to respond to intramuscular vaccine. AB - OBJECTIVE: To study the humoral immune responses, safety, and tolerability of intradermal recombinant hepatitis B vaccination in healthcare workers (HCWs) nonresponsive to previous repeated intramuscular vaccination. DESIGN: An open, prospective, before-after trial. SETTING: A tertiary referral hospital and surrounding district health service in Queensland, Australia. PARTICIPANTS: Hospital and community HCWs nonresponsive to previous intramuscular hepatitis B vaccination. METHODS: Intradermal recombinant hepatitis B vaccine was administered every second week for a maximum of 4 doses. Hepatitis B surface antibody (anti-HBs) responses were assessed 2 weeks after each dose. RESULTS: Protective anti-HBs levels developed in 17 (94%) of 18 study subjects. Three doses resulted in seroconversion of all responding subjects and the highest geometric mean antibody concentration. The vaccine was well tolerated. CONCLUSION: More than 90% of previously nonresponsive HCWs responded to intradermal recombinant hepatitis B vaccine with protective anti-HBs levels. PMID- 11894842 TI - Artificial cation-conducting channels: design, synthesis, and characterization. AB - A nonpeptidic, cation-conducting channel has been designed, synthesized, and evaluated. The channel was found to conduct protons and Na+ through phospholipid bilayers. The evaluation of Na+ transport was conducted using a dynamic 23Na-NMR (nuclear magnetic resonance) in phospholipid vesicles and a planar bilayer using a patch clamp amplifier. Several control compounds were prepared to determine which of the structural "modules" were necessary. Experiments using fluorescent residues and fluorescence energy transfer were undertaken to locate the channel within the bilayer and to demonstrate that the channel functions as a monomeric unit. PMID- 11894843 TI - Effects of tumors on inhaled pharmacologic drugs: I. Flow patterns. AB - Lung carcinomas are now the most common form of cancer. Clinical data suggest that tumors are found preferentially in upper airways, perhaps specifically at carina within bifurcations. The disease can be treated by aerosolized pharmacologic drugs. To enhance their efficacies site-specific drugs must be deposited selectively. Since inhaled particles are transported by air, flow patterns will naturally affect their trajectories. Therefore, in Part I of a systematic investigation, we focused on tumor-induced effects on airstreams, in Part II (the following article [p. 245]), particle trajectories were determined. To facilitate the targeted delivery of inhaled drugs, we simulated bifurcations with tumors on carinas using a commercial computational fluid dynamics (CFD) software package (FIDAP) with a Cray T90 supercomputer and studied effects of tumor sizes and ventilatory parameters on localized flow patterns. Critical tumor sizes existed; e.g., tumors had dominant effects when r/R > or = 0.8 for bifurcation 3-4 and r/R > or = 0.6 for bifurcation 7-8 (r = tumor radius and R = airway radius). The findings suggest that computer modeling is a means to integrate alterations to airway structures caused by diseases into aerosol therapy protocols. PMID- 11894840 TI - Effect of geldanamycin on androgen receptor function and stability. AB - In the ligand-binding inactive state, the steroid receptor heterocomplex contains Hsp90, Hsp70, high-molecular weight immunophilins, and other proteins. Hsp90 acts in association with co-chaperones to maintain the native state of the receptor within the cells. It was reported earlier that Hsp90 might not be as important for the androgen receptor (AR) activity as for the glucocorticoid receptor (GR) and the progesterone receptor (PR) activities. We used the Hsp90 inhibitor geldanamycin (GA) to explore the role of Hsp90 in the function of the AR heterocomplex. GA selectively binds to Hsp90 and inhibits its activity, leading to the loss of steroid receptor activity, and frequently, its degradation. In our study, LNCaP prostate cancer cells were treated with GA for 30 minutes or 24 hours, in the presence of mibolerone, a synthetic androgen. GA reduced the androgen-induced AR protein levels to 15% after 24 hours of treatment. Several androgen up-regulated genes, including immunophilin FKBP51 and prostate specific antigen (PSA), were reduced by GA treatment. In cells treated with GA after transfection with a PSA promoter or an androgen response element-driven reporter gene, AR-mediated transactivation of reporter gene expression was reversibly inhibited by GA. Loss of androgen-binding ability and AR levels was attributed to reduced transcription of AR-regulated gene expression. Degradation rate of 35S labeled AR was significantly increased by GA in the presence or absence of mibolerone. GA induced the degradation of AR through the proteasomal pathway. AR in cells treated with proteasomal inhibitor lactacystin, was insoluble in Nonidet P-40 (NP40)-based buffer and could not restore the androgen-binding ability. We report here that GA treatment disrupted both hormone-binding activity and receptor protein stability, resulting in a dramatic loss of androgen-induced gene activation. These results show that Hsp90 activity is important for both the chaperone-mediated folding of the AR into a high-affinity ligand-binding conformation and the functional activity of the AR. PMID- 11894844 TI - Effects of tumors on inhaled pharmacologic drugs: II. Particle motion. AB - Computer simulations were conducted to describe drug particle motion in human lung bifurcations with tumors. The computations used FIDAP with a Cray T90 supercomputer. The objective was to better understand particle behavior as affected by particle characteristics, airflow conditions, and disease-modified airway geometries. The results indicated that increases in particle sizes, breathing intensities and tumor sizes could enhance drug deposition on the tumors. The modeling suggested that targeted drug delivery could be achieved by regulating breathing parameters and designing (selecting physical features of) aerosolized drugs. We present the theoretical work as a step towards improving aerosol therapy protocols. Since modeling describes factors affecting dose, it is complementary to considerations of the molecular aspects of drug formulation and pharmacokinetics. PMID- 11894845 TI - Three-dimensional computer modeling of the human upper respiratory tract. AB - Computer simulations of airflow and particle-transport phenomena within the human respiratory system have important applications to aerosol therapy (e.g., the targeted delivery of inhaled drugs) and inhalation toxicology (e.g., the risk assessment of air pollutants). A detailed description of airway morphology is necessary for these simulations to accurately reflect conditions in vivo. Therefore, a three-dimensional (3D) physiologically realistic computer model of the human upper-respiratory tract (URT) has been developed. The URT morphological model consists of the extrathoracic (ET) region (nasal, oral, pharyngeal, and laryngeal passages) and upper airways (trachea and main bronchi) of the lung. The computer representation evolved from a silicone rubber impression of a medical school teaching model of the human head and throat. A mold of this ET system was sliced into 2-mm serial sections, scanned, and digitized. Numerical grids, for use in future computational fluid dynamics (CFD) simulations, were generated for each slice using commercially available software (CFX-F3D), AEA Technology, Harwell, UK. The meshed sections were subsequently aligned and connected to be consistent with the anatomical model. Finally, a 3D curvilinear grid and a multiblock method were employed to generate the complete computational mesh defined by the cross-sections. The computer reconstruction of the trachea and main bronchi was based on data from the literature (cited herein). The final unified 3D computer model may have significant applications to aerosol medicine and inhalation toxicology, and serve as a cornerstone for computer simulations of air flow and particle-transport processes in the human respiratory system. PMID- 11894846 TI - Cellular function and control of volume-regulated anion channels. AB - Restoration of cell volume after cell swelling in mammalian cells is achieved by the loss of solutes (K+, Cl-, and organic osmolytes) and the subsequent osmotically driven efflux of water. This process is generally known as regulatory volume decrease (RVD). One pathway for the swelling induced loss of Cl- (and also organic osmolytes) during RVD is the volume-regulated anion channel (VRAC). In this review, we discuss the physiological role and cellular control of VRAC. We will first highlight evidence that VRAC is more than a volume regulator and that it participates in other fundamental cellular processes such as cell proliferation and apoptosis. The second part concentrates on the Rho/Rho kinase/myosin phosphorylation cascade and on compartmentalization in caveolae as modulators of the signal transduction cascade that controls VRAC gating in vascular endothelial cells. PMID- 11894847 TI - Caldesmon and smooth-muscle regulation. AB - Smooth muscles exist in the wall of hollow organs in our body and are responsible for controlling the flow of vital fluids that are essential for the normal function of the cardiovascular, respiratory, digestive, and reproductive systems. Many diseases, such as hypertension, asthma, indigestion, and premature birth, may attribute to malfunction of smooth-muscle contraction. It is therefore important to decipher how smooth-muscle contraction is regulated. This review attempts to give a brief overview of current understanding about the molecular mechanisms of smooth-muscle regulation and, in particular, to discuss possible roles of caldesmon in this regulatory process. PMID- 11894848 TI - Replication-related activities establish cohesion between sister chromatids. AB - Replicated sister chromatids are held together from their synthesis in S phase to their separation in anaphase. The process of sister chromatid cohesion is essential for the proper segregation of chromosomes in eukaryotic cells. Recent studies in Saccharomyces cerevisiae have advanced our understanding of how sister chromatid cohesion is established, maintained, and dissolved during the cell cycle. Historical observations have suggested that establishment of cohesion is roughly coincident with replication fork passage. Emerging evidence now indicates that replication fork components, such as PCNA, a novel DNA polymerase, Trf4p/Pol sigma (formerly Trf4p/Pol kappa), and a modified clamp-loader complex, actively participate in the process of the cohesion establishment. Here, we review the molecular events in the chromosome cycle with respect to cohesion. Failure of sister chromatid cohesion results in the aneuploidy characteristic of many birth defects and tumors in humans. PMID- 11894849 TI - Stimulation of lipid peroxidation in vitro in rat brain by the metabolites accumulating in maple syrup urine disease. AB - In this study we investigated the in vitro effects of the metabolites accumulating in maple syrup urine disease on lipid peroxidation in brain of young rats. Chemiluminescence and thiobarbituric acid-reactive substances were measured in brain homogenates from 7- and 30-day-old rats in the presence of 10 mM of the branched-chain amino acids L-leucine, L-isoleucine, or L-valine; their keto acids L-2-ketoisocaproic acid, L-2-keto-3-methylvaleric acid, or L-2-ketoisovaleric acid; or the hydroxy derivatives L-2-hydroxyisocaproic acid, L-2-hydroxy-3 methylvaleric acid, or L-2-hydroxyisovaleric acid separately added to the incubation medium. We observed that all amino acids, keto acids, and hydroxy acids accumulating in this disease stimulate to a variable degree the in vitro parameters of lipid peroxidation tested in homogenates of rat brain. The results indicate a possible participation of oxidative stress in the neuropathology of maple syrup urine disease patients, especially during a crisis, when the metabolites are highly increased, and point to the use of antioxidant drugs as a possible adjuvant therapy in such situations to improve the neurological status of the patients and to prevent sequelae. PMID- 11894850 TI - Traces left on visual selective attention by stimuli that are not consciously identified. AB - Briefly presented information, even if unidentifiable, may speed or delay required responses to following events. It has been assumed that this priming of the motor system may occur without affecting attention to priming and following stimuli. In contrast to this notion, the present study reports that such unidentified stimuli have effects on a physiological indicator of the attentional system. A lateral posterior electroencephalogram component was evoked by laterally presented relevant shapes, reflecting shifts of attention to those shapes. This component was absent, however, when the relevant shape was preceded by a similar shape at the same location, even if this shape was completely masked by metacontrast. The attentional shift evidently became unnecessary in this situation. Thus, unidentifiable information may leave some trace for attention controlled selection of the following event. PMID- 11894852 TI - From the Centers for Disease Control and Prevention. Laboratory-acquired meningococcal disease--United States, 2000. PMID- 11894853 TI - From the Centers for Disease Control and Prevention. Barriers to dietary control among pregnant women with phenylketonuria--United States, 1998-2000. PMID- 11894851 TI - Very happy people. AB - A sample of 222 undergraduates was screened for high happiness using multiple confirming assessment filters. We compared the upper 10% of consistently very happy people with average and very unhappy people. The very happy people were highly social, and had stronger romantic and other social relationships than less happy groups. They were more extraverted, more agreeable, and less neurotic, and scored lower on several psychopathology scales of the Minnesota Multiphasic Personality Inventory. Compared with the less happy groups, the happiest respondents did not exercise significantly more, participate in religious activities significantly more, or experience more objectively defined good events. No variable was sufficient for happiness, but good social relations were necessary. Members of the happiest group experienced positive, but not ecstatic, feelings most of the time, and they reported occasional negative moods. This suggests that very happy people do have a functioning emotion system that can react appropriately to life events. PMID- 11894855 TI - JAMA patient page. Living with sarcoidosis. PMID- 11894854 TI - From the Centers for Disease Control and Prevention. Lyme disease--United States, 2000. PMID- 11894856 TI - What's new in plastic surgery. PMID- 11894857 TI - Laparoscopic splenectomy has become the gold standard in children. AB - Splenectomy is frequently required in children with various hematologic disorders. The reported advantages of laparoscopic splenectomy (LS) include less pain, shorter hospital stay, and improved cosmesis. This report evaluates the outcome of children undergoing LS at a single children's facility. One hundred twelve children underwent LS by the lateral approach between August 1995 and February 2001. Indications for LS were hereditary spherocytosis in 58, idiopathic thrombocytopenic purpura in 21, sickle cell disease in 19, and other conditions in 14. LS alone was completed in 89 children and LS and cholecystectomy (LSC) in 20. Three required conversion to open splenectomy. Accessory spleens were identified in 19. Complications included ileus (four), acute chest syndrome (four), bleeding (two), pneumonia (one), and diaphragm perforation (one). There was no mortality. An accessory spleen was missed in one child with recurrent anemia. Average operative time for LS was 106 minutes and for LSC 135 minutes. Operative time for LS decreased with experience but the difference was not significant. Average length of stay was 1.51 days (range 1-11) and was longer in sickle cell disease (2.47 days) versus hereditary spherocytosis (1.29 days) and idiopathic thrombocytopenic purpura (1.16 days). We conclude that LS is safe and effective in children with hematologic disorders and is associated with minimal morbidity, zero mortality, and a short length of stay. PMID- 11894858 TI - Laparoscopic transabdominal preperitoneal inguinal hernia repair for recurrent inguinal hernia. AB - Laparoscopic transabdominal preperitoneal inguinal herniorrhaphy (TAPP) was attempted on 989 hernias in 708 patients. Of these 137 (14%) hernias were recurrent after conventional anterior repair. Laparoscopic repair was performed on 135 recurrences in 120 patients (the study cohort). There were 119 males and three females with an average age of 59 years (22-83 years). One hundred twelve (83%) were the first recurrences, and 23 (17%) were multiple recurrences. In 90 recurrences (66.7%) the last repair was performed more than 10 years previously. Seventy-seven patients (64%) had a prior or concomitant repair of a contralateral hernia. Direct recurrences and right-side recurrences were more common (73% and 61% respectively). Postoperative complications occurred in 18 repairs (13%). These included 15 hematomas, two seromas, and one urinary retention. Re recurrence occurred in one patient (0.7%) in whom hernia staples were not used. No recurrence has occurred since the hernia staples became available. We conclude that the overall incidence of recurrent inguinal hernia is high (14%). Patients with recurrent hernia have a tendency toward a contralateral hernia (64%). Most recurrences occurred 10 or more years after the previous repair (66.7%). The laparoscopic repair (TAPP) offers a good repair for recurrent inguinal hernia avoiding the scar tissue and with low complication and recurrence rates. PMID- 11894859 TI - The philosophical foundations of Darwinism. PMID- 11894860 TI - Social benefits of a successful biomedical research company: Merck. PMID- 11894861 TI - Charity and strategy: philanthropy's evolving role. PMID- 11894863 TI - Abstracts of the British Pharmacological Society Meeting. 5-7 September 2001. PMID- 11894862 TI - International consensus statement on malignant peripheral nerve sheath tumors in neurofibromatosis. AB - Neurofibromatosis 1 (NF1) is an autosomal dominant tumor predisposition syndrome in which affected individuals have a greatly increased risk of developing malignant peripheral nerve sheath tumors (MPNSTs). These cancers are difficult to detect and have a poor prognosis. Because patients may present to specialists from widely differing disciplines, the association with NF1 is often not appreciated, and there is no cohesive pattern of care. A multidisciplinary group of 33 clinicians and scientists with specialist knowledge in MPNST and NF1 reviewed the current published and unpublished data in this field, and distilled their collective experience to produce a consensus summary on MPNST in NF1. The known clinical, pathological, and genetic information on MPNST in NF1was collated, and a database was established to record information in a uniform manner. Subgroups with a higher risk of developing MPNSTwere identified within the NF1 population. The consortium formulated proposals and guidelines for clinical and pathological diagnosis, surgical management, and medical treatment of MPNST in individuals with NF1.A multidisciplinary team approach to the management of this complex disorder is advocated. Progress can be made by adopting the guidelines proposed by this consortium and by widespread dissemination of standardized information. Collaborative research should be promoted with the aim of harnessing advances in molecular genetics to develop targeted therapies for MPNST in people with NF1. PMID- 11894864 TI - Abstracts of the 46th annual meeting of the German Society of Endocrinology. Gottingen, February 27-March 2, 2002. PMID- 11894865 TI - Abstracts of the British Geriatrics Society autumn meeting. 18-19 October 2001. London, United Kingdom. PMID- 11894866 TI - Effects of sound preconditioning on hearing loss from low or middle-frequency noise exposure. AB - Objective. To explore prior noise exposures or sound conditioning as a moderator of hearing loss produced by traumatic exposure to low or middle-frequency noise. Method. Two experimental groups of guinea-pigs were conditioned using a 0.5 kHz octave band noise (OBN) at 85 dB, 6 h/d for 4 d. The subjects were allowed to recover for 3 d after conditioning. Then the first group was exposed to a 0.5 kHz OBN at 110 dB for 1 h, the second group was exposed to 1 kHz OBN at 110 dB for 1 h. Two control groups received 0.5 kHz and 1 kHz OBN respectively at 110 dB for 1 h without prior sound conditioning. Result. Hearing threshold shifts recorded at 48 h after the high-level noise exposure in conditioned groups and control groups demonstrated that conditioning provided significant protection on hearing threshold shift from low or middle frequency noise exposure. Histological examination revealed significantly less hair cell loss in the conditioned than in the control groups. In addition, malondialdehyde (MDA) levels in red blood cell (RBC) of conditioned groups were significantly lower as compared with that of the control groups. Conclusion. Low-frequency conditioning provided significant protection not only on hearing threshold shift caused by noise of the same frequency, but also on that caused by middle frequency noise. PMID- 11894867 TI - [Age dependency of heart rate variability, blood pressure variability and baroreflex sensitivity]. AB - Objective. To compare the cardiovascular autonomic regulatory function between young and middle-aged male subjects and to assess the effects of aging. Method. Spectral indices of short term heart rate variability (HRV), systolic blood pressure variability (SBPV), and baroreflex sensitivity (BRS) in both supine and upright positions were obtained by AR spectral analysis and the sequential method, respectively, for both groups. Result. HRV spectral indices in both supine and upright, and the BRS in supine were lower in the middle-aged subjects than in the young subjects (P< 0.05, or P< 0.01). From supine to upright position total power (TP), low frequency power (LF) and high frequency power (HF) of HRV spectrum did not show any significant changes in the middle-aged subjects, whereas in the young subjects typical changes were observed. SBPV spectral indices did not show significant differences between the two groups in both supine and upright positions. However, SBPV HF was an exception, being lower in supine position in the middle-aged than in the young subjects (P< 0.01). For the young subjects, in supine position, BRS was correlated with LF and HF of HRV, respectively, and the spectral indices of HRV in supine position were also correlated with their corresponding indices in upright position. For the middle aged subjects these correlations did not exist. Conclusion. In the middle-aged, both HRV and BRS are reduced and the correlations between HRV indices and BRS are not existent. However, the short-term blood pressure variability seems to be not age-dependent. It suggests that different regulatory mechanisms might account for it and that under resting condition the feed forward regulation from HRV to SBPV might be unimportant. PMID- 11894868 TI - [Effects of different training methods on cardiovascular autonomic regulation during bedrest]. AB - Objective. To investigate the effects of different training methods on cardiovascular autonomic regulation under bedrest. Method. 15 healthy male volunteers aged 19-22 participated tests in head-down tilt (HDT) -6 degrees bedrest in order to observe the changes of cardiovascular system under simulated weightlessness. They were divided into control (5 men), hypoxia training (5 men) and Fangsong training (5 men) groups. 24 h dynamic ECG were recorded on the 2nd day of pre-bedrest, on the 3rd, 14th and 18th day of bedrest and on the 7th day of the post- bedrest. All spectra were estimated from entire 24 h HRV, before, during and after Fangsong and hypoxia training by autoregressive (AR) modeling method. Normalized low-frequency (LF%) was a quantitative marker of cardiac sympathetic activity, normalized high-frequency (HF%) reflected the changes in cardiac vagal activity, and LF/HF was considered to be related to sympathovagal balance or sympathetic activity. Result. In control group, LF% and HF% were all significantly reduced (P<0.05), LF/HL showed no significant changed during bedrest. In Fangsong group, HF% increased markedly (P< 0.05), while in hypoxia group, LF% increased markedly (P< 0.05). Conclusion. Fangsong training counteracted markedly the reduction in vagal activity, while hypoxia training counteracted markedly the decrease in sympathetic activity. It was possible that HRV indices could be used to evaluate the efficiency of countermeasures counteracting the adverse effects of weightlessness. PMID- 11894869 TI - [Effects of intermittent +45 degrees head-up-tilt on mechanical parameters of femoral (correction of femural) bone in tail-suspended rats]. AB - Objective. To study the effects of different intermittent +45 degrees head-up tilt (IHUT) on tail-suspended rats. Method. 28 male SD rats were randomly and equally divided into: control group (C), tail-suspended group (S), IHUT 2 h group (H2), and IHUT 4 h group (H4). On the basis of tail-suspension to simulate weightlessness, H rats were given 2 h or 4 h IHUT each day. The effects of 3 wk IHUT on tail-suspended rats were evaluated by measuring physical and mechanical parameters of the femur. Result. Femoral mass (fresh, dry and ash), diameter, and density (fresh, except H2 group) in S, H2 and H4 rats were declined significantly compared to C rats (P<0.01 or P<0.05). Diameter (P<0.05) and density of femur (P<0.01) in H2 rats were significantly improved compared to S rats. Compared to C rats, the strength and stiffness of femora were obviously weakened (P<0.01) in S rats. Elastic load in H2 rats improved markedly (P<0.05) than those in S group; in H4 group, elastic load, maximum load, and bending toughness coefficient improved significantly (P< 0.01 or P<0.05). Conclusion. Using IHUT, mechanical parameters of weight bearing bones of tail-suspended rats can be improved apparently, by extending the exposure time, mechanical parameters can be improved. PMID- 11894870 TI - [Human physiological regulation in the closed suit with no ventilation]. AB - Objective. To study the effects of the Experimental Closed Suit (ECS) with no active ventilation on human physiological functions and its anti-asphyxic role. Method. Five young healthy male subjects, aged 18-21 years, were tested in the ECS with no active ventilation and no oxygen supply (NAVANOS) when the gas regulatory valve switch was placed on A or B position respectively. Result. (1) Under 1 atm air pressure, each subject could endure at least 15 min while they were aware and resting in the ECS with NAVANOS; (2) There was a balancing period or plateau of the oxygen and the carbon dioxide pressure in the vicinity of the mouth in 5 to 7 min after the sealed helmet was closed, and this period lasted to the end of each test; (3) During the 8-10 min balancing period, the oxygen partial pressure in the vicinity of the mouth ranged from 15.0 to 16.4 kpa, which was slightly hypoxic, and the range of the carbon dioxide partial pressure was 5.21 to 6.49 kPa, which was within the CO2 concentration-time curve of human physiological endurance; (4) No significant difference in human physiological parameters was found when the regulatory valve was placed on A or B position. Conclusion. The anti-asphyxic time of ECS with NAVANOS could at least last 15 min under 1 atm air pressure while the subjects were aware and resting. PMID- 11894871 TI - [A study on establishing method in analysing EEG state complexity]. AB - Objective. To establish method in analysing EEG complexity as related to brain function state (Cs) and to examine Cs under conditions of taking rest and doing mental arithmetic, to explore their regular changes. Method. EEG of 10 male subjects aged 26-30 under the conditions of taking rest and doing mental arithmetic were recorded. Cs under the two conditions and their changes related to brain function states were analyzed with a special software for Cs analysis. Result. EEG Cs under mental arithmetic changed significantly. According to these changes, the subjects were divided into two groups: Group A, 8 subjects, their EEG Cs under mental arithmetic were significantly higher than Cs under rest at all electrode sites. The most outstanding areas were temporal region and parieto occipital region. Group B, 2 subjects, their EEG Cs under mental arithmetic were significantly lower than Cs under rest. The most outstanding areas were prefrontal region and temporal region. Conclusion. The result showed that high correlation existed between mental arithmetic and Cs. This method of analysis was useful in studying brain cognitive activities and assessing mental workload. PMID- 11894874 TI - [A heat transfer model for liquid cooling garment (LCG) and its analysis]. AB - Objective. To establish the heat transfer model for liquid cooling garment (LCG) and, basing on this model, to find the relations between the design parameters and heat removing, as well as that between the design parameters and heat transfer efficiency. Method. Heat transfer process of the LCG was analyzed according to engineering facts. Result. The relations between the design parameters and heat removing, and also that between the design parameters and heat transfer efficiency were interelative and the optimal values of the parameters were essential to the design of LCG. Conclusion. The results might be useful in the design of LCG in extra vehicular activity (EVA) space suit. PMID- 11894873 TI - [The development of a static water/gas separator]. AB - Objective. To develop a device for separating water from gas in small flow rate under zero "G". Method. Beginning with the study of surface characteristic of materials, a capillary material was developed according to the requirement and the water/gas separator using this single separating material was designed. Result. The water/gas separator worked well in the range of gas flow below 10.0 L/min and water flow below 10.0 ml/min. No gas was found in the separated water and no water was found in the separated gas. Conclusion. The structure of the separator was reasonable and the water/gas separating method using a single separating material was feasible. PMID- 11894872 TI - [Development of an experimental facility for waste treatment by microorganism]. AB - Objective. To construct an experimental facility for microorganism waste processing, which will be used to recover plant nutrient liquids from plant inedible biomass essential for growth and development of plants. Method. After technical parameters and performance requirements were defined, plan demonstration, drawing design, fabrication, debug and preliminary plant inedible residue-biodegradation tests of microorganisms were conducted. Result. The temperature, stirring speed and gas-supplying flow of bioreactor of the facility were controlled automatically, as well as the pH and dissolved oxygen concentration were measured automatically and controlled manually, testifying that its performance reached the requirements of predetermined technical indexes. The 15-d test showed that the facility ran smoothly, its above-mentioned parameters could be measured and controlled precisely, and the biodegradation rate of lettuce's inedible biomass approximately attained 90%. Conclusion. The facility holding reasonable technical indexes, smooth and dependable performances, is capable of being utilized to biodegrade plant inedible biomass. It is expected that if the above-mentioned parameter combinations are optimized further, the results should be better. PMID- 11894875 TI - [Implementation of a software for acquiring and analysing myoelectric potential signals in ergonomical research of space manual system]. AB - Objective. A software used for acquiring and analyzing signals was developed for ergonomical research on Human Workload in space manual system. Method. As an important part of the whole experimental equipment and being developed in PC, the software is composed of acquisition and display program, analysis and processing program and data files. Result. The software is capable of making realtime acquisition and display of four channels of myoelectric potential signals and one channel of operation signal synchronously. The signals are then analyzed and processed off-line. Conclusion. During the development, its realtime feature was implemented by means of some technical methods, such as that different machine assigned to view-scenery display and signal acquisition, different frequency assigned to EMG signal and operation ones, and different cycle assigned to acquisition, display and storage. Above all, with friendly man-machine interface and high accuracy of data, the software was even reliable. PMID- 11894876 TI - [Changes of mRNA expression of IL-1beta and TNF-alpha in rat brains after repeated exposures to +Gz]. AB - Objective. To study changes of mRNA expression of IL-1beta and TNF-alpha in rat brains after repeated exposures to +Gz. Method. Twenty conscious SD rats served as the subjects. They were randomly divided into 5 groups. Using an animal centrifuge, control rats (n = 4) were exposed to +1 Gz and experimental rats (n = 16) were exposed to +14 Gz three times, each for 45 s with 30 min interval in between. The rat brains were taken 30 min, 6 h, 24 h and 48 h after the last centrifuge run and total RNA was isolated. mRNA expression levels of IL-1beta and TNF-alpha in rat brain were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Result. mRNA expression levels of IL-1beta and TNF-alpha in rat brains taken 30 min, 6 h and 24 h after repeated +Gz exposures were significantly higher than those in control rats, but returned to normal after 48 h. Conclusion. It suggested that mRNA expressions of IL-1beta and TNF-alpha in rat brains can be stimulated by repeated +Gz exposures and the increased expression of IL-1beta and TNF-alpha may play a role in the pathologic course of brain damage induced by +Gz exposures, but the damage is reversible. PMID- 11894878 TI - [Incidence and prevention of space decompression sickness]. AB - To expound the necessity that space decompression sickness (SDCS) should be separated from altitude DCS, comparison was made between the features of space DCS and altitude DCS. The etiology and pathogenesis of the space DCS and altitude DCS were the same, but the features of the rules leading to their incidences (included influencing factors etc.) were different. For the convenience of getting better systemic and definite knowledge about SDCS, and making effective preventive plans and theoretical expositous, SDCS should be taken as an independent professional term. PMID- 11894877 TI - [Changes of glucocorticoid receptor in cerebral and hepatic cytosol during decompression stress injury in rats]. AB - Objective. To observe the changes of glucocorticoid receptor (GR) in cerebral and hepatic cytosol during decompression stress injury in rats. Method. 30 rats were divided into 5 group. They were placed into the compression chamber for compression and decompression. The binding capacity of GR of cerebral and hepatic cytosol were measured by the exchange assay, using 3H dexamethasone as the ligand. Meanwhile, decompression bubbles on pericardial area were measured using Doppler ultrasonic method. Result. The binding capacity of GR of cerebral and hepatic cytosol reduced after decompression stress injury in the animals, especially cerebral cytosol (P<0.01, P<0.05). The result also showed that the binding capacity of cerebral and hepatic GR should have further decreased, if the therapeutic measure had not been used in animals suffered from decompression sickness (DCS). Conclusion. The changes of the binding capacity of GR of cerebral and hepatic cytosol were proved to be related to decompression stress injury, which might be taken as one of the indices for evaluating injury degree of DCS. PMID- 11894879 TI - [Ground-based studies on thermoregulation at simulated microgravity by head-down tilt bed rest]. AB - Comparisons of headward fluid shifts, plasma volume losses, altered vasodilator and vasoconstrictor responses, reduced exercise capacities, etc. between microgravity and head-down tilt (HDT) bedrest suggested that HDT model was suitable for simulating thermoregulation under microgravity. Both microgravity and simulated microgravity by HDT exposures degraded human thermoregulation ability in some aspects. The features of the responses to heat stress on men after HDT exposures were higher increase rate in rectal temperature, attenuated increase rates of skin temperature and body heat conductance, less heat dissipation from core to skin and higher sensitivity of sweating etc. PMID- 11894880 TI - [Changes of cerebral circulation during weightlessness or simulated weightlessness]. AB - The results about studies on changes of the cerebral circulation during weightlessness/simulated weightlessness were reviewed in this paper. The possible influencing mechanism of weightlessness on cerebral circulation and its physiological significance were summarized. It could be concluded that the changes of cerebral circulation were the results of self-regulation of the brain to maintain its normal function, and it might play an important role in the genesis of postflight orthostatic intolerance. PMID- 11894882 TI - Engendering sacrifice: blood, lineage, and infanticide in old French literature. PMID- 11894881 TI - Sine rege, sine principe: Peter the Venerable on violence in twelfth-century Burgundy. PMID- 11894883 TI - Fleet Market to Bloomsbury: T.N.R. Morson's progress. PMID- 11894884 TI - European pharmacies in Colonial India. PMID- 11894885 TI - The Chicago Five: a family group of integrative psychobiologists. AB - The "Chicago Five" was a group of integrative psychobiologists, including Frank A. Beach, Donald O. Hebb, David Krech, Norman R. F. Maier, and Theodore C. Schneirla, all of whom worked with Karl S. Lashley at the University of Chicago during 1929 -1935. Although they went on to careers in diverse fields of psychology, their approaches reflect a set of underlying themes that can be traced to their experiences in Chicago. Nine primary beliefs that, with occasional exceptions, underlie their work are delineated. The term family is proposed to refer to a group of psychologists who share a common professional development in one place within a limited time period and whose later work although it may be diverse, reflects commonalities that may be traced to that experience. PMID- 11894886 TI - The course in the history of psychology: present status and future concerns. AB - The present study provides a turn-of-the-century status report on the teaching of the history of psychology in colleges and universities in the United States. The data indicate that the course is offered regularly in most departments of psychology and is frequently required of majors; these findings are consistent with earlier research. Most instructors teach the course largely out of personal interest and self-taught expertise with their primary teaching and research commitments to other areas of psychology. Few instructors engage in publication of research and scholarship in the history of psychology, although there are 2 journals in the field that provide an outlet for scholarship. The few positions that allow for primary commitment to teaching and research in the history of psychology is a possible cause of concern for the future of the course and for its place in the education of psychologists in the 21st century. PMID- 11894887 TI - From reassurance to irrelevance: adolescent psychology and homosexuality in America. AB - American psychology by the 1920s contained a greater capacity for viewing some homosexual experiences as normal than most current historical literature suggests. Developmental psychologists agreed with psychiatrists that adult homosexuality was pathological, but they also agreed that adolescent sexual development included a homosexual phase. Until the late 1960s, developmental texts reassured parents and teachers that homosexual behavior among adolescents was transitory and quite normal. The psychiatric view of homosexuality as pathology came under attack after the middle of the century and eventually was abandoned. The developmental concern with a transitory homosexual phase disappeared gradually. This trend in psychology suggests underlying social and cultural changes. PMID- 11894888 TI - Computer automated approach to the extraction of epiphyseal regions in hand radiographs. AB - Epiphyseal region is the most sensitive region to developmental changes of the skeletal system. Extraction of this area is the very first step in any computerized image analysis. In this report a fully automated analysis of a hand radiograph resulting in extraction of distal and middle regions of the II, III, and IV phalanx is presented. The processing is performed in 3 stages. First, the trend of background is removed from radiograph to obtain a binary hand mask. At this stage a labeling procedure is necessary to eliminate artifacts (markers). Then, II, III, and IV phalanges are identified in the binary image, and the phalangeal axes are drawn. Finally, the intensity profile along each phalangeal axis is analyzed, and, on its basis, distal and middle regions are located. The presented procedure is designed as a part of currently developed system for automatic bone age assessment; however, it also can be as a preprocessing step in other diseases the diagnoses of which may require a computer assistance. PMID- 11894889 TI - Semiautomatic segmentation of the cochlea using real-time volume rendering and regional adaptive snake modeling. AB - The human cochlea in the inner ear is the organ of hearing. Segmentation is a prerequisite step for 3-dimensional modeling and analysis of the cochlea. It may have uses in the clinical practice of otolaryngology and neuroradiology, as well as for cochlear implant research. In this report, an interactive, semiautomatic, coarse-to-fine segmentation approach is developed on a personal computer with a real-time volume rendering board. In the coarse segmentation, parameters, including the intensity range and the volume of interest, are defined to roughly segment the cochlea through user interaction. In the fine segmentation, a regional adaptive snake model designed as a refining operator separates the cochlea from other anatomic structures. The combination of the image information and expert knowledge enables the deformation of the regional adaptive snake effectively to the cochlear boundary, whereas the real-time volume rendering provides users with direct 3-dimensional visual feedback to modify intermediate parameters and finalize the segmentation. The performance is tested using spiral computed tomography (CT) images of the temporal bone and compared with the seed point region growing with manual modification of the commercial Analyze software. Our method represents an optimal balance between the efficiency of automatic algorithm and the accuracy of manual work. PMID- 11894890 TI - Thresholding histogram equalization. AB - The drawbacks of adaptive histogram equalization techniques are the loss of definition on the edges of the object and overenhancement of noise in the images. These drawbacks can be avoided if the noise is excluded in the equalization transformation function computation. A method has been developed to separate the histogram into zones, each with its own equalization transformation. This method can be used to suppress the nonanatomic noise and enhance only certain parts of the object. This method can be combined with other adaptive histogram equalization techniques. Preliminary results indicate that this method can produce images with superior contrast. PMID- 11894891 TI - Image display for collision avoidance of radiation therapy: treatment planning. AB - Patient treatment in a medical linear accelerator is characterized by many angular and translational movements of the gantry and couch. The direction and orientation of each treatment beam is specified by a set of gantry, turntable, and collimator angles. It is possible that some selected treatment field configurations will result in gantry/couch or gantry/patient collisions that remain undetected during the treatment planning process. In this work, a digital camera has been used to record all the workable gantry/ patient set-up images, and a Windows programming language is used to edit and display these images on a personal computer for the treatment planner to screen the treatment plans. These graphical displays enable the planner to be aware of any potential collision hazards by an actual visualization of each selected gantry/turntable or gantry/patient angle configuration. PMID- 11894892 TI - An automated results notification system for PACS. AB - The purpose of this study was to determine if the interval between an examination being ordered by an Emergency Department physician and his or her review of the report and images could be shortened by notifying the physician that the results were available. This hypothesis was based on work done previously in the Medical Intensive Care Unit that showed that physicians would wait to review results for a time considerably longer than the time required for the radiologist to review the images and provide a preliminary report. The software developments operate properly and show that even simple integration of multiple information systems (PACS, RIS, speech recognition) can provide useful features. Early results indicate that the Emergency Department (ED) physicians prefer the notification system over the previous (travel to check on images and reports) methods. The hypothesized time reductions did occur, although it is not clear that the notification system accounted for all of them. A system for automated notification of radiology results availability has been shown to be possible and practical. To do this automated interaction of 3 systems with a low-level or no electronic integration was required. Although not fully successful for this study, early physician response has been positive, and requests to expand this service hospitalwide now are common. PMID- 11894893 TI - Computerized scientific exhibit utilization: observations from infoRAD at the radiologic society of North America Scientific Assembly. AB - No publication has discussed utilization of computer scientific exhibits (CSE) at national symposia, despite their growing numbers. The hypothesis of this project was that, when given a choice, viewers initially would prefer a more conventional paper presentation of a scientific exhibit over that of an electronic presentation. A nearly identical paper version of the introductory screen to an infoRAD CSE was placed adjacent to the workstation. Utilization of the paper introduction, computer introduction, and both, as well as subsequent behavior, was recorded. Of 67 visitors, initial user choice was 56.7% paper and 43.3% computer. Over the entire time at the exhibit 25.4% only looked at the handout, 25.4% only at the computer, and 49.3% perused both. Only 10.5% completed the entire exhibit, and 0.94% of total registrants visited the CSE. Overall, 74.7% perused the CSE when leaving the exhibit area. Upon arrival, viewers preferred the more conventional paper presentation, confirming the project hypothesis. Surprisingly, about 75% eventually perused at least a portion of the computer presentation. Although a small fraction of Radiologic Society of North America (RSNA) registrants visited the CSE, the findings presented are promising and suggest that CSE presence at national meetings is justifiable, providing a "first step" toward CME outcomes analysis of CSE. Overall, these findings are promising and suggest that computer scientific exhibit presence at national meetings is justifiable. PMID- 11894894 TI - The cloning and expression of the gene encoding organ-specific esterase S from the genome of Drosophila virilis. AB - We have cloned the gene for the esterase S isozymes complex from the genome of Drosophila virilis in pBR322. Esterase S is an enzyme which is specifically synthesized in the ejaculatory bulbs of D. virilis adult males. The gene for the esterase S isozyme complex (estS) has been localized in band 2G5e of chromosome II. Poly(A)+ RNA prepared from ejaculatory bulbs actively hybridizes with this band. A cloned 15-kb fragment of D. virilis DNA (pVE9) also hybridizes with band 2G5e. The area encoding the poly(A)+ RNA is located in the middle part of the cloned fragment whose ends are not transcribed in vivo. Only one poly(A)+ RNA which is 1.9 kb long and complementary to pVE9 DNA can be revealed in the cytoplasm. The mRNA preselected by hybridization to pVE9 DNA was microinjected into the cytoplasm of Xenopus laevis oocytes. In other experiments, the pVE9 DNA itself was microinjected into oocyte nuclei. In both cases, esterase S is synthesized in the oocytes, and the major part of the protein is transported from the oocytes and accumulated in the incubation medium. PMID- 11894895 TI - The topology of the proton translocating F0 component of the ATP synthase from E. coli K12: studies with proteases. AB - The accessibility of the three F0 subunits a, b and c from the Escherichia coli K12 ATP synthase to various proteases was studied in F1-depleted inverted membrane vesicles. Subunit b was very sensitive to all applied proteases. Chymotrypsin produced a defined fragment of mol. wt. 15,000 which remained tightly bound to the membrane. The cleavage site was located at the C-terminal region of subunit b. Larger amounts of proteases were necessary to attack subunit a (mol. wt. 30,000). There was no detectable cleavage of subunit c. It is suggested that the major hydrophilic part of subunit b extends from the membrane into the cytoplasm and is in contact with the F1 sector. The F1 sector was found to afford some protection against proteolysis of the b subunit in vitro and in vivo. Protease digestion had no influence on the electro-impelled H+ conduction via F0 but ATP-dependent H+ translocation could not be reconstituted upon binding of F1. A possible role for subunit b as a linker between catalytic events on the F1 component and the proton pathway across the membrane is discussed. PMID- 11894896 TI - Biosynthesis of caerulein in the skin of Xenopus laevis: partial sequences of precursors as deduced from cDNA clones. AB - The decapeptide caerulein represents one of the main constituents of the skin secretion of Xenopus laevis. Total mRNA was isolated from skin, transcribed into cDNA and inserted via GC-tailing into the plasmid pUC8. Among the transformants, 300 clones were selected at random and screened with a cDNA primed with the synthetic deoxynucleotide d(AGTCCATCCA), which is complementary to the mRNA region coding for the fragment Trp-Met-Asp-Phe of cerulein. Of nine strongly hybridizing clones, three were sequenced and these were found to contain inserts with very similar nucleotide sequences. The cloned cDNAs code for parts of two different caerulein precursors. These contain one or two copies of caerulein and five additional amino acids located between pairs of arginine residues. The extra glycine at the carboxy terminus is considered to serve as the signal for amidation, while the tetrapeptide Phe-Ala-Asp-Gly, linked to the amino end of caerulein in these precursors, must be cleaved by an unusual processing mechanism. PMID- 11894897 TI - Monoclonal antibody to human 66,000 molecular weight plasminogen activator from melanoma cells. Specific enzyme inhibition and one-step affinity purification. AB - Hybridomas producing a monoclonal IgG1 antibody to a human plasminogen-activating enzyme with an apparent mol. wt. of 66,000 (66 K, HPA66) from human melanoma cells were obtained by fusion of NSI-Ag 4/1 mouse myeloma cells with spleen cells from a mouse immunized with a partially purified preparation of the enzyme. Screening for clones of hybridomas producing antibodies to HPA66 was performed with the impure enzyme preparation. A preliminary screening included enzyme linked immunosorbent assay and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting; the final identification was based on inhibition of the enzymatic activity of HPA66 which was complete at high antibody concentrations. No inhibition of three other human and murine plasminogen activators or of plasmin was observed. Employing a one-step affinity procedure with the antibody coupled to Sepharose, HPA66 was purified approximately 200-fold from conditioned medium from the melanoma cells with a yield of 79%. The purified HPA66 was homogeneous as evaluated by SDS-PAGE. Electrophoresis under reducing conditions indicated that it consisted of one polypeptide chain. The binding constant between the antibody and 125I-labelled HPA66 was approximately 2.5 x 10(9) l/mol. The antibody did not bind to a variety of other plasminogen activators, including 52-K and 36-K human enzymes and 48-K and 75-K murine enzymes. Previously, a monoclonal antibody against another enzyme was derived by the sole use of enzyme inhibition for screening. The present study represents a modification of this procedure that can be used when antibody-unrelated inhibitors of the enzyme are present in hybridoma culture fluid. PMID- 11894898 TI - Polymorphism and complexity of the human DC and murine I-A alpha chain genes. AB - A cDNA clone encoding the human B cell alloantigen DC alpha chain (pDCH1) has been used to analyse the structure of the human and murine major histocompatibility complexes by the DNA filter hybridization technique. The pDCH1 probe hybridizes to a single DNA sequence present on chromosome 17 in the mouse genome. A restriction enzyme polymorphism enables us to map this sequence to the I-A subregion. Extensive restriction enzyme polymorphism detected in HLA-DR homozygous typing cells is reminiscent of the DR/MT linkage disequilibrium groups, suggesting that the pDCH1 probe could be useful for haplotype typing in the human population. The HLA-DR region appears more complex than the I region since a second DC-like hybridizing sequence is detected in the human genome in these experiments. PMID- 11894900 TI - The mammalian primase is part of a high molecular weight DNA polymerase alpha polypeptide. AB - The recently discovered eukaryotic primases have been found in tight association with certain DNA polymerase alpha forms. Here I present evidence that the high mol. wt. catalytic polypeptide (125,000) of an apparently homogeneous DNA polymerase alpha from freshly harvested calf thymus contains both polymerase and primase activity. This conclusion derives from the following three facts: (1) the two enzyme activities cannot be separated upon velocity sedimentation in 1.7 M urea, (2) both activities elute at a pI of 5.25 upon chromatofocussing and (3) after SDS-electrophoresis, renaturation of the enzymes in situ and measurement of DNA polymerase and primase activities in the gels, both enzymes have identical mobilities and coincide with the high mol. wt. catalytic subunit of DNA polymerase alpha. PMID- 11894901 TI - The DNA-protein cross: a method for detecting specific DNA-protein complexes in crude mixtures. AB - A method using crude cellular mixtures is described which permits identification of polypeptides and DNA fragments forming specific complexes. Our procedure incorporates elements of both 'Southern' and 'protein' blotting and combines, in two dimensions, the resolving power of a denaturing protein gel with that of an agarose DNA gel. Conditions for 'crossing' have been established using the lambda repressor-operator system: the specific complex can be detected by crossing total protein from bacteria overproducing the repressor with a mixture of total genomic fragments from a lysogen. PMID- 11894899 TI - Sequence and expression of the mouse mammary tumour virus env gene. AB - We have determined the DNA sequence of the envelope gene region of the GR strain of mouse mammary tumour virus. The sequence extends for 3012 nucleotides from the single EcoRI site to beyond the PstI site in the 3' long terminal repeat (LTR) of the provirus. There is a major open reading frame from nucleotides 752 to 2818 which encompasses the entire env gene. This reading frame extends through a polypurine tract and into the LTR. There is another open reading frame from the first nucleotide to position 803, presumably corresponding to the end of the pol gene. The splice acceptor site which generates env mRNA has been mapped experimentally to nucleotide 750. The env gene products, gp52 and gp36, have been positioned on the sequence using the directly determined amino acid sequences of the amino terminus of gp52; and both the amino and carboxyl termini of gp36. The start of gp52 is preceded by a series of 19 uncharged amino acids which could function as a typical signal sequence, but this sequence is only part of a much longer leader peptide. The tetrad Arg-Ala-Lys-Arg is the presumed cleavage site in the gPr73env precursor, and occurs just before the gp36 amino terminus. There are five potential asparagine-linked glycosylation sites which agrees with previous experimental results. The gp36 has two long hydrophobic regions at its amino and carboxy termini, these are suggested to act as a fusion peptide and the trans-membrane anchor, respectively. PMID- 11894902 TI - Evidence for a coupling of synthesis and export of an outer membrane protein in Escherichia coli. AB - We describe a lesion, lamB701-708, affecting the hydrophilic portion of the lambda receptor signal sequence. The C to A transversion of the sixth codon of the signal sequence changes a positively charged arginine to a neutral serine. The phenotype conferred by this alteration is unique among previously described signal sequence mutations. The results suggest an essential role for the charged amino acids of the hydrophilic segment in the initial interaction between a nascent secreted protein and a membrane export site. The results further suggest that synthesis of lambda receptor is coupled to its export. PMID- 11894903 TI - Late transient expression of human hepatitis B virus genes in monkey cells. AB - The expression of human hepatitis B virus (HBV) surface (HBS) and e (HBe) antigens has been studied comparatively in monkey and mouse cell lines co transfected with HBV DNA and the dominant selective marker aminoglycoside 3' phosphotransferase gene. We have found that the kinetics and stability of expression of the HBS gene varies with the cell lines used. Only a late transient expression of both HBS and HBe is observed between 1 and 5 weeks after transfection in monkey kidney Vero cells transfected with the complete HBV genome, while a permanent expression of HBS and HBe is obtained in mouse cells. HBS and HBe are excreted into the cell culture medium. HBe is expressed in cells transfected with the complete HBV genome, but not with isolated HBS gene. In clones of Vero cells transformed with the HBS gene, HBV sequences were rearranged or lost. PMID- 11894904 TI - Binding between the par region of plasmids R1 and pSC101 and the outer membrane fraction of the host bacteria. AB - The binding between par+ and par plasmid DNA to different membrane fractions of Escherichia coli was investigated. Membrane material from cells carrying different Par+ and Par- derivatives of plasmids R1 and pSC101 was isolated and fractionated into an outer and a cytoplasmic membrane fraction. The presence of plasmid DNA in the two membrane fractions was measured either by nick-translation of the membrane-bound DNA, followed by filter-hybridization to homologous DNA, or by filter-hybridization of the membrane-bound DNA to nick-translated homologous purified plasmid DNA. The DNA of par derivatives of plasmids R1 and pSC101 could be detected only in the cytoplasmic membrane fraction, whereas the corresponding par+ plasmid DNA also appeared in the outer membrane material, indicating a specific binding between the R1 and pSC101 partition loci and the bacterial outer membrane. The experiment was then modified by fractionation of the membrane material from cells carrying hybrids between the vector pSF2124 and the par region or the basic replicon region of plasmid R1. The DNA of the membrane fractions were filter-hybridized to nick-translated probes. Again, the par+ region caused hybridization to the outer membrane material. Therefore, we may conclude that controlled partitioning involves binding of DNA to membrane material that has the same density as the outer membrane of the host bacteria. This finding offers a biochemical 'assay' for studies of the molecular biology of plasmid partitioning. PMID- 11894905 TI - Conservation of RNA secondary structures in two intron families including mitochondrial-, chloroplast- and nuclear-encoded members. AB - Two families of fungal mitochondrial introns that include all known sequences have been recognized. These families are now extended to incorporate a plant mitochondrial intron and several introns in chloroplast- and nuclear-encoded rRNA and tRNA precursors. Members of the same family share distinctive sequence stretches and a number of potential RNA secondary structures that would bring these stretches and the intron-exon junctions into relatively close proximity. Using several of these introns which have been extensively studied by either biochemical or genetic means, an attempt is made to integrate the available data into a common picture. PMID- 11894906 TI - Nitrogen control of the nif regulon in Klebsiella pneumoniae: involvement of the ntrA gene and analogies between ntrC and nifA. AB - The ntrC and nifA gene products of Klebsiella pneumoniae are transcriptional activators involved in general nitrogen control and nif-specific regulation, respectively. Multicopy plasmids expressing either ntrC or nifA from a foreign promoter were used to study the relationship between these two genes and ntrA. The nifA product substituted for ntrC product in activation of a number of genes including nifLA, hutUH and genes for arginine and proline utilisation. NtrC could not substitute for nifA in transcriptional activation of the nifHDKY operon. In ntrA- strains, neither the ntrC nor the nifA product functioned to activate transcription of nif promoters. In vitro transcription/translation studies with plasmid clones demonstrated similar levels of expression of ntrC and nifA in ntr+ and ntrA- S-30 extracts. Hence, lack of activator function in an ntrA mutant indicates that both the ntrC and nifA products require a functional ntrA for activity. When expressed from foreign promoters, both the ntrC and nifA products were active in conditions which would normally repress nif expression. Hence, the ntrA product was apparently not limiting in these conditions. PMID- 11894907 TI - Isolation and characterization of a collagen-binding glycoprotein from chondrocyte membranes. AB - A collagen-binding glycoprotein was isolated from purified chick chondrocyte surface membranes by affinity chromatography on type II collagen-Sepharose. The purified glycoprotein has an apparent mol. wt. of 31,000 and binds to native chick collagen types I, II, III, V and M. Although it contains 30% carbohydrates, the majority of which is fucose, it is hydrophobic and soluble only in detergents. The integral membrane protein character of the 31-K protein became apparent from its ability to insert into lecithin vesicles. Liposome-inserted 31 K protein binds 125I-labelled type II collagen in the presence of 0.5 M NaCl, while detergent-solubilized 31-K protein is dissociated from type II collagen by 0.05-0.1 M NaCl. Electron microscopic studies employing the rotary shadowing technique indicate that 31-K protein particles bind to the ends of collagen molecules. We propose that this glycoprotein serves as anchorage site for extracellular collagen to the chondrocyte membrane and thus may be involved in cell-matrix interactions in cartilage. PMID- 11894908 TI - Organization of internucleosomal DNA in rat liver chromatin. AB - A detailed analysis of the length distribution of DNA in nucleosome dimers trimmed with exonuclease III and S1 nuclease suggests that the previously described variation of internucleosomal distance in rat liver occurs, at least for a subset of the nucleosomes, by integral multiples of the helical repeat of the DNA. Results obtained upon digestion of chromatin with DNase II further suggest that lengths of internucleosomal DNA are integral multiples of the helical repeat of the DNA plus approximately 5 bp. Restraints imposed by these features on the arrangement of nucleosomes along the fiber are discussed. PMID- 11894909 TI - Cloning of several cDNA segments coding for human liver proteins. AB - A human cDNA library was constructed using M13 derivative vectors. The simple and rapid procedures for sequencing single-stranded DNA by the dideoxy chain termination method allowed a screening of individual clones directly by DNA sequence analysis. Some of these clones were identified as coding for: serum albumin, alpha1-antitrypsin, retinol-binding protein, prothrombin, haptoglobin, and metallothionein. Furthermore, a clone coding for aldolase B was tentatively identified on the basis of high sequence homology with rabbit muscle aldolase. PMID- 11894910 TI - Internal structural anisotropy of spherical viruses studied with magnetic birefringence. AB - Six so called spherical viruses (four plant and two animal) are shown to exhibit magnetically induced birefringence in solution. They must therefore be magnetically and optically anisotropic. This is attributed to static structural anisotropy of the interiors as neither natural shape nor field-induced deformations are likely causes. Thus at least part of these virus cores have a symmetry differing from that of their capsids. An estimate of the average orientation of the RNA bases is given for the plant viruses: turnip yellow mosaic, bromegrass mosaic, tomato bushy stunt and turnip crinkle. The packing geometry of the nucleic acid/protein cores of adenovirus and probably influenza virus are anisotropic but to an extent that cannot be quantified. PMID- 11894911 TI - IS2 insertion is a major cause of spontaneous mutagenesis of the bacteriophage P1: non-random distribution of target sites. AB - Insertion mutations arising spontaneously in the P1 prophage and affecting vegetative phage reproduction were screened for the presence of insertion sequence 2 (IS2). Filter hybridization identified 28 out of 44 independent insertions as IS2. Their target specificity is not random. A region that amounts to < 2% of the phage genome had trapped 15 of the 28 IS2 elements. However, precise mapping of nine mutants in this hot spot segment revealed no preferred insertion site. Rather, the nine IS2 are distributed over the whole target segment and IS2 are found in both orientations. Sequence data indicate that at least two sequence variants of IS2 participated in mutagenesis of the phage genome. The detectable transposition of IS2 from the host chromosome to the prophage occurs with a frequency of 3 x 10(-5) per cell per generation under the particular experimental conditions. It is concluded that IS2, a natural resident of Escherichia coli K12 strains, is an important agent for spontaneous mutagenesis and exerts this action non-randomly along the genome. PMID- 11894912 TI - The E1b promoter of Ad12 in mouse L tk- cells is activated by adenovirus region E1a. AB - We have investigated the effect of the E1a region of adenovirus on the expression of the E1b transcriptional unit in mouse L tk- cells. To that end we have fused the promoter region of the E1b gene of Ad12 to the coding sequence of the thymidine kinase gene of herpes simplex virus type 1. We found that expression of this fusion gene is completely dependent on the presence of the E1a region. Sequences involved in this regulation were mapped between positions -135 and +11 from the E1b cap site. In addition, we found that the largest E1a protein is essential in this regulation process. PMID- 11894913 TI - Genetic study of a membrane protein: DNA sequence alterations due to 17 lamB point mutations affecting adsorption of phage lambda. AB - Gene lamB encodes the outer membrane receptor for phage lambda in Escherichia coli K12. We have determined the DNA sequence alterations of 17 lamB point mutations which result in resistance to phage lambda h+. The mutations correspond to four phenotypic classes according to the pattern of growth of three phages which use the lambda receptor: lambda h (a one-step host-range derivative of lambda h+), lambda hh* (a two-step host-range derivative of lambda h+) and K10 (another lambdoid phage). Fourteen mutations are of the missense type and correspond to Gly to Asp changes distributed as follows. One class I mutation is at position 382 of the mature lambda receptor. Seven class I* mutations, four of which at least are independent, are at position 401. Six independent class II mutations are at position 151. The three other (class III) mutations are of the nonsense type. They change codons TGG (Trp) into TAG (amber) at positions 120 (two mutations) and 351 (one mutation). Implications of these results for the topological organization of the lambda receptor as well as possible reasons for the limited number of altered sites detected are discussed. PMID- 11894914 TI - Negative dominance in gene lamB: random assembly of secreted subunits issued from different polysomes. AB - lamB is the structural gene for the lambda receptor, an oligomeric outer membrane protein from Escherichia coli K12 involved in phage lambda adsorption. We show that, under certain conditions, in a strain diploid for gene lamB, all the missense lamB mutations conferring lambda resistance that we have tested are dominant with respect to wild-type. We propose a model which allows a quantitative interpretation of the data. It is based on negative complementation at the level of oligomerisation. Wild-type and mutant subunits would assemble at random forming homo- and hetero-oligomers. Only wild-type homo-oligomers would be efficient for phage inactivation. For some classes of missense mutations the hetero-oligomers would have the capacity to bind, but not to inactivate the phage. The model confirms that active lambda receptor is a trimer and implies that for this secreted protein there is no preferential assembly of subunits originating from the same polysome. PMID- 11894915 TI - Prolipoprotein signal peptidase of Escherichia coli requires a cysteine residue at the cleavage site. AB - A signal peptidase specifically required for the secretion of the lipoprotein of the Escherichia coli outer membrane cleaves off the signal peptide at the bond between a glycine and a cysteine residue. This cysteine residue was altered to a glycine residue by guided site-specific mutagenesis using a synthetic oligonucleotide and a plasmid carrying an inducible lipoprotein gene. The induction of mutant lipoprotein production was lethal to the cells. A large amount of the prolipoprotein was accumulated in the outer membrane fraction. No protein of the size of the mature lipoprotein was detected. These results indicate that the prolipoprotein signal peptidase requires a glyceride modified cysteine residue at the cleavage site. PMID- 11894916 TI - Differences among the polyadenylated RNA sequences of human leucocyte populations: an approach to the objective classification of human leukaemias. AB - We have constructed a complementary DNA (cDNA) library representing expressed sequences of the white blood cells from a patient with chronic granulocytic leukaemia. The library was screened by colony hybridization of 32P-labelled cDNAs synthesized from the polyadenylated RNAs of the white blood cells from patients with chronic granulocytic or chronic lymphocytic leukaemia. The autoradiographic patterns were compared and 70 recombinants were selected to comprise a panel which distinguished between these two types of leukaemia. Hybridization of this panel with complementary DNAs transcribed from the polyadenylated RNAs of a variety of normal and neoplastic leucocyte populations showed that the RNA sequences in high abundance in leucocytes from chronic granulocytic leukaemias differ quite radically from those in other leucocytes. The patterns of hybridization seen when this panel was challenged with cDNAs representing the RNAs of normal and leukaemic leucocyte populations were sufficiently different to distinguish clearly the peripheral blood leucocytes of chronic granulocytic leukaemias from other populations of white blood cells, both normal and leukaemic. We suggest that this approach might provide additional markers useful in the classification of the acute leukaemias, especially the undifferentiated leukaemias whose identification by conventional methods is uncertain. PMID- 11894917 TI - Post-transcriptional control of expression of the repA gene of plasmid R1 mediated by a small RNA molecule. AB - Fusions between the repA gene of plasmid R1 (required for autonomous plasmid replication) and the lac genes have been the basis for in vivo studies of regulation of repA expression. Two gene products--the CopA RNA and the CopB protein--act as inhibitors of repA expression. Comparison of the effects of addition in trans of the two Cop functions on transcription and translation of repA-lac gene fusions show that the CopB protein represses transcription of the repA gene, whereas the CopA RNA interferes with the RepA mRNA in such a way that effective translation is inhibited. The CopA RNA does not seem to have a direct effect on the transcription of the repA gene but, as a consequence of the posttranscriptional regulation, transcriptional polarity within the repA gene is observed. It is also shown that the CopA RNA interacts with its target (CopT) only when the region is transcribed to form RepA mRNA. PMID- 11894918 TI - Membrane integration and function of the three F0 subunits of the ATP synthase of Escherichia coli K12. AB - Integration into the cytoplasmic membrane and function of the three F0 subunits, a, b and c, of the membrane-bound ATP synthase of Escherichia coli K12 were analysed in situations where synthesis of only one or two types of subunits was possible. This was achieved by combined use of atp mutations and plasmids carrying and expressing one or two of the atp genes coding for ATP synthase subunits. AU three F0 subunits were found to be required for the establishment of efficient H+ conduction. Subunits a and b individually as well as together were found to bind F1 ATPase to the membrane while subunit c did not. The ATPase activity bound to either of these single subunits, or in pairwise combinations, was not inhibited by N,N'-dicyclohexylcarbodiimide. Also ATP-dependent H+ translocation was not catalysed unless all three F0 subunits were present in the membrane. The integration into the membrane of the subunits a and b was independent of the presence of other ATP synthase subunits. PMID- 11894920 TI - Total purification of a DNA-dependent ATPase and of a DNA-binding protein from human cells. AB - We have purified to near homogeneity the major DNA-dependent ATPase from human cells. The pure enzyme has a mol. wt. of 68,000 and a minimum specific activity of approximately 150 U/mg. When the properties of the pure enzyme are compared with those of a less purified preparation, significant differences are observed both in structure and in function. These can be ascribed to the interaction of the ATPase with a DNA-binding protein (mol. wt. 28,000) that we can also purify to near homogeneity from the same cells and which is present in the less purified preparations of the ATPase. The ability of the less purified ATPase to stimulate DNA polymerase alpha in helicase fashion is probably due to the presence of the DNA-binding protein. PMID- 11894919 TI - Dibutyryl cAMP treatment of neuroblastoma-glioma hybrid cells results in selective increase in cAMP-receptor protein (R-I) as measured by monospecific antibodies. AB - The absolute levels of cAMP-dependent protein kinase (cAMP-dPK) subunits (R-I, R II and C) and cGMP-dependent protein kinase (cGMP-dPK) holoenzyme were studied in neuroblastoma-glioma hybrid cells before and after dibutyryl-cAMP (Bt2cAMP) treatment which results in differentiation of these cells. The levels were determined by two different techniques utilizing antibodies which had been raised against each individual purified protein kinase subunit (or the holoenzyme in the case of the cGMP-dPK). Electrophoretic transfer of samples from SDS polyacrylamide gels to nitrocellulose paper, followed by immunolabeling of protein kinase subunits with their respective antibodies and [125I]Protein A, demonstrated the monospecific nature of the antibodies, and a selective, several fold increase in the R-I subunit in Bt2cAMP-treated cells, with no change in the level of R-II or C subunits. A simple enzyme-linked immunosorbent assay (ELISA) capable of measuring nanogram amounts of the various subunits confirmed the selective increase in the R-I subunit. ELISA assay results also indicated that the R-I subunits present before and after Bt2cAMP treatment are antigenically homologous. In conclusion, the specific, sensitive immunological methods described here demonstrate the capacity of neuroblastoma-glioma hybrid cells to regulate separately the levels of the two distinct subunits (R-I and C) of the Type I cAMP-dPK. PMID- 11894921 TI - Simultaneous expression of mu- and gamma-chain mRNA in cloned murine B-lymphoma cell lines. AB - The expression of immunoglobulin heavy chain genes was studied in five murine B lymphomas known from previous studies to express either mu (38C-13), mu + delta (L10A, K46, BCL1) or gamma chains (A20). The presence of mu- and gamma-mRNAs in these tumors was determined by Northern blot analyses of the total cell poly(A)+ mRNA, using the appropriate 32P-labeled recombinant plasmid probes. In four out of the five lymphomas examined, both mu- and gamma-mRNAs were detected. The mu mRNA appeared as multiple discrete bands of 1.9-3.0 kb. In three out of the four lymphomas, the gamma-mRNA appeared as two bands, a major one of 1.9 and a minor one of 3.9 kb. Three myelomas examined by similar methods did not contain more than one class of heavy chain mRNA. Reexamination of the Ig chains produced by the B-lymphomas which expressed both mu- and gamma-mRNAs revealed that two of them preserved their original phenotype and expressed mu (38C-13) or gamma chains only (A20). In contrast, two of the cell lines previously shown to express mu but not gamma chains (i.e., L10A and K46R) had changed during growth in culture and 'switched' to the production of gamma chains only. These results indicate that, in contrast to myelomas, B-lymphomas possess two classes of mRNA. However, the production of heavy chain mRNA in B-lymphomas is not necessarily accompanied by synthesis of the corresponding polypeptide chains. More studies are necessary to find out whether the expression of 'non-productive' heavy chain mRNA molecules in B-lymphomas is related to the phenomena of 'allelic exclusion' and/or the 'heavy chain switch' which occurs during the maturation of B-cells. PMID- 11894922 TI - Spontaneous high expression of heat-shock proteins in mouse embryonal carcinoma cells and ectoderm from day 8 mouse embryo. AB - When submitted to a heat-shock, mouse embryonal carcinoma (EC) and fibroblast cells show very different behavior. All the EC cells so far analyzed express very high levels of several heat-shock proteins (HSP) in the absence of stress and independent of their origin and culture conditions. In such cells, the 89-kd, 70 kd and 59-kd HSP are the most prominent proteins after actin. In addition, the 89 kd and 59-kd HSP are not stimulated by an arsenite shock in contrast to what is observed with fibroblasts or cells of the parietal yolk sac type. Arsenite induces the synthesis of a 105-kd polypeptide in fibroblasts but not in EC cells. In vitro differentiation of F9 cells induced by retinoic acid and dibutyryl cAMP is accompanied by a decrease in the spontaneous relative abundance of HSP and restores the arsenite-induced synthesis of the 105-kd polypeptide. EC cells are usually believed to be similar to inner cell mass cells of mouse blastocyst. Furthermore, data in the literature together with our own results suggest that the same three HSP are also spontaneously expressed in high amounts in the early mouse embryo. PMID- 11894923 TI - Pausing of RNA polymerase molecules during in vivo transcription of the SV40 leader region. AB - Viral transcription complexes were isolated from SV40-infected cells and incubated in vitro in the presence of [alpha-32P]UTP to allow elongation of the promoter-proximal RNA up to the attenuation sites. The 94 nucleotide attenuated RNA (spanning nucleotides 243-336) was purified, digested with RNase T1 and fingerprinted. The labeled oligonucleotides were then isolated, digested with RNase T2 and their base composition was determined. Based on these analyses 10 consecutive oligonucleotides, spanning residues 259-336, were identified. As the in vivo synthesized oligonucleotides are unlabeled the junctions between labeled and unlabeled oligonucleotides define the in vivo pause sites of RNA polymerase molecules. The characterization of the 10 radioactive spots and their relative intensities allowed the localization of two in vivo pause sites: one at 13-16 nucleotides downstream from the major initiation site presumably at the initial opening of the DNA helix and the second at approximately 40 nucleotides downstream from the major initiation site, just past a GC-rich region of dyad symmetry. It is postulated that pausing of RNA polymerase molecules in the leader region is an essential process in the control of SV40 late transcription. PMID- 11894924 TI - In vitro premature termination in SV40 late transcription. AB - Nuclear extracts and viral transcribing minichromosomes were prepared from SV40 infected cells and incubated in vitro with [alpha-32P]UTP under conditions which allow the elongation of preinitiated RNA chains. Sucrose gradient lysis of the transcription mixtures revealed two populations of SV40-specific RNA: elongating chains that remain associated with the viral minichromosomes, and, at the top of the gradient, small free RNA detached from the template and hybridizing exclusively to the promoter-proximal region of SV40 DNA. This free RNA was shown by polyacrylamide gel electrophoresis to comprise essentially a 94 nucleotide species, which could, however, at high UTP concentration, be elongated a further few nucleotides before terminating. These results thus show that the actively transcribing minichromosomes provide a sytem in which the attenuated RNA can be released from the template. Moreover, this is the first demonstration of specific in vitro termination of polymerase B transcription. The conditions which lead to transcription termination are discussed. PMID- 11894925 TI - Anti-schistosome monoclonal antibodies of different isotypes--correlation with cytotoxicity. AB - Five monoclonal antibodies specific towards Schistosoma mansoni antigens were prepared by fusion of spleen cells of infected and immunized mouse with the murine myeloma NS-1 cells. Three of the five antibodies belonged to the IgG1 class, one was an IgM and the fifth one was an IgE. The IgE monoclonal antibody designated 54.10, induced antigen-specific degranulation of rat basophilic cell line, a property which served as the basis for the screening assay. Its biological function was demonstrated by a specific macrophage activation that led to killing of schistosomula; no such killing was obtained with anti-schistosome antibodies of other classes or with IgE of different antigenic specificity. The second monoclonal antibody of biological significance was an IgG1, designated 27.21 which is reactive in the immunofluorescence staining of surface antigens on intact schistosomula. All three monoclonal antibodies that belonged to the IgG1 class were effective in mediating killing of schistosomula by complement, with the highest effect exerted by 27.21. It is thus apparent that the 27.21 monoclonal antibody is directed against a densely distributed surface antigen on the schistosomula membrane which is possibly involved in the protective immunity. Preliminary data showed that immunoprecipitation with the 27.21 antibodies results in the isolation of three major protein bands, of 60 kd, 50 kd, 19 kd, respectively. PMID- 11894926 TI - Induction of oligodendrocyte-like properties in a primitive hypothalamic cell line by cholesterol, an eye derived growth factor and brain extract. AB - A serum-free medium has been devised which permits proliferation of the mouse primitive nervous cell line F7. When cholesterol, eye-derived growth factor and brain extract are added in this medium for 48 h, 80-90% of oligodendrocyte-like cells are generated. These cells have diminished substrate adhesion. They acquire the capacity to synthesize carbonic anhydrase II and myelin basic protein, two specific proteins of oligodendrocytes. These observations suggest that F7 clonal cell line, which has been previously shown to be a neurophysin cell precursor, is also a precursor for oligodendrocytes, and represents a bipotent stem cell line for both neuronal and glial cell lineages. PMID- 11894927 TI - The promoter region of the arg3 gene in Saccharomyces cerevisiae: nucleotide sequence and regulation in an arg3-lacZ gene fusion. AB - We have determined the DNA sequence for the 5' end of the arg3 gene of Saccharomyces cerevisiae, including part of the coding region and the 200 nucleotides immediately upstream. A promoter-deletion mutant was found to have lost all of the sequence lying normally in front of the gene except for the 33 nucleotides preceding the AUG codon. The role of the 5' domain in initiation and regulation of arg3 transcription was assessed by a gene fusion experiment. The Escherichia coli lacZ gene, was truncated of the eight amino-terminal codons substituted in vitro, on a 2mu plasmid, for the carboxy-terminal and 3'-flanking regions of arg3, leaving only the first 19 proximal codons and approximately 1600 nucleotides of the region preceding arg3 on the yeast chromosome. The fused gene was expressed in phase and was still submitted to the two mechanisms regulating the wild-type arg3 gene: the general, probably transcriptional control of amino acid biosynthesis and the specific, apparently post-transcriptional control mediated by the products of the argR genes. These results suggest a determining role for the 5' end portion of the arg3 messenger in the specific arginine mediated control mechanism. PMID- 11894928 TI - Fusion of DNA region to murine immunoglobulin heavy chain locus corresponds to plasmacytoma-associated chromosome translocation. AB - Murine plasmacytomas frequently exhibit a translocation of the distal region of chromosome 15 to the end of chromosome 12, where the immunoglobulin heavy chain locus resides. A candidate for the DNA across the chromosome fusion point is a cloned region of non-immunoglobulin DNA which in most plasmacytomas has recombined near the alpha heavy chain constant region gene. That the incoming DNA, provisionally designated LyR (lymphoid rearranging) DNA, does derive from chromosome 15 is shown here by blot analysis of DNA from two panels of somatic cell hybrids: hybridomas between an AKR T-lymphoma (Tikaut) and CBA mouse cells with a cytogenetically distinctive chromosome 15, and between mouse and Chinese hamster cells. LyR DNA segregated with chromosome 15 in all lines and the results assign LyR to the distal two thirds of that chromosome. This assignment, together with the previously reported high frequency of recombination between LyR and C(alpha) in plasmacytomas and associated alteration of LyR transcription suggests that translocation activates a LyR gene involved in plasmacytoma oncogenesis. Moreover, LyR rearrangement in certain T-lymphomas, such as the Tikaut line examined here, also implicate that gene in oncogenesis of some T-lymphomas. PMID- 11894929 TI - Comparison of the binding sites for the Escherichia coli cAMP receptor protein at the lactose and galactose promoters. AB - Polyacrylamide gel electrophoresis has been used to visualise and quantitate complexes between the Escherichia coli cyclic AMP receptor protein (CRP) and DNA fragments containing the promoter region of either the E. coli galactose or lactose operons. We show that, although CRP binding to the gal fragment is weaker than binding to the lac fragment, in each case, stable complexes are formed between one dimer of CRP and one molecule of DNA. We have examined the effects of a series of deletions and point mutations in the gal promoter region on CRP binding. From the position of deletions and mutations which prevent the formation of stable complexes, we deduce the location and extent of the sequence at the CRP binding site. We show that it covers approximately the same length of sequence as the binding site at the lac promoter. Unlike the lac site, the gal site contains no palindromic sequence. We discuss the importance of symmetry in the sequence at CRP binding sites and the validity of CRP binding consensus sequences which have been proposed. PMID- 11894930 TI - Molecular characterization of the human red cell Rho(D) antigen. AB - Human red cells of Rh blood groups -D-/-D- ('super-D'), -/- (Rhnull) and normal Rho(D)+ cells were radioactively surface-labeled using the lactoperoxidase 125I method. Polyacrylamide gel electrophoresis in the presence of SDS followed by fluorography showed a strong enrichment of a polypeptide with an apparent mol. wt. of 28,0000-33,000 in the 125I-labeled -D-/-D- membranes. This polypeptide was specifically immune precipitated with anti-Rho(D) antiserum. Treatment of intact cells with trypsin or Pronase did not digest the protein. The Rho polypeptide migrated identically on polyacrylamide gel electrophoresis under reducing and non reducing conditions. It was not phosphorylated after in vitro incubation of red cells with 32P. When whole labeled membranes were solubilized in neutral detergent and applied to lectin-Sepharose columns the Rho(D) polypeptide adsorbed to Ricinus communis lectin but not to wheat germ lectin or Lens culinaris lectin. The purified molecule did not adsorb to R. communis lectin-Sepharose. Treatment of the Rho(D) antigen with endo-N-acetyl glucosaminidase H, endo-beta galactosidase or mild alkali did not lower its apparent mol. wt. PMID- 11894931 TI - Stable transformation of mouse teratocarcinoma stem cells with the dominant selective marker Eco.gpt and retention of their developmental potentialities. AB - Transformation of PCC4 mouse teratocarcinoma stem cells was obtained using a dominant selective marker, the enzyme xanthine-guanine phosphoribosyltransferase (XGPRT), coded by the bacterial Eco.gpt gene placed under the control of the early SV40 genes in the vector pSV2gpt. An average of 20 colonies of transformed cells was obtained, using the calcium phosphate technique, 10 microg DNA vector, no carrier DNA and 1 x 10(6) recipient cells. Five independent Eco.gpt transformed PCC4 cell lines were propagated in selective medium and assayed for XGPRT activity. All of them had the ability to convert [14C]xanthine to xanthine monophosphate. pSV2gpt sequences were present and associated with high mol. wt. cellular DNA. pSV2gpt sequences and XGPRT activity were both conserved in the three clones that were propagated in non-selective medium for 30 generations. The transformed PCC4 cells retained their ability to produce, in host mice, teratocarcinoma tumors composed of embryonal carcinoma and various differentiated tissues. Thus, pSV2gpt can be used as a dominant marker to select teratocarcinoma stem cells co-transformed with genes that are not selectable by themselves. PMID- 11894932 TI - Antibody-linked polymerase assay on protein blots: a novel method for identifying polymerases following SDS-polyacrylamide gel electrophoresis. AB - We describe a method for correlating polymerase activity with a particular polypeptide band in an SDS-polyacrylamide gel which does not require renaturation of the SDS-denatured enzyme. The method involves the following steps: (i) transfer of proteins from an SDS-polyacrylamide gel onto nitrocellulose; (ii) incubation with excess antiserum raised against a partially purified polymerase preparation to link one Fab site of an antibody molecule to the denatured enzyme on the nitrocellulose; (iii) binding of native polymerase to the other Fab site of the antibody molecule in the immune complex to generate a specific polymerase 'sandwich'; (iv) assaying of the nitrocellulose filter for antibody-linked native polymerase activity using an appropriate template and a radioactive substrate followed by treatment with trichloroacetic acid to precipitate in situ the radioactive product. The essential feature of this method is that the use of both non-specific anti-polymerase serum and a partially purified enzyme preparation is sufficient to allow identification of a specific protein following SDS polyacrylamide gel electrophoresis. This antibody-linked polymerase assay has been developed to identify a 130,000-dalton RNA-dependent RNA polymerase from cowpea leaves. Possible applications of this type of assay as a tool for identifying a wide variety of proteins are discussed. PMID- 11894933 TI - The beta2-microglobulin mRNA in human Daudi cells has a mutated initiation codon but is still inducible by interferon. AB - The human Burkitt lymphoma cell line Daudi does not synthesize beta2 microglobulin (beta2m) and lacks the cell surface histocompatibility antigens. The cells, however, contain RNA hybridizing to a cloned human beta2m cDNA probe. cDNA from this Daudi beta2m RNA, was cloned and sequenced. By comparison with cDNA prepared from Ramos cells, which synthesized microglobulin, we determined the sequence of the 20 amino acid long leader peptide of pre-beta2m and show that in Daudi cells the initiator ATG has been mutated to ATC. Although Daudi beta2m RNA cannot be translated, interferon induces the beta2m RNA in Daudi cells as well as in normal human cells. PMID- 11894934 TI - Mouse histocompatibility genes: structure and organisation of a Kd gene. AB - The gene coding for the mouse H-2Kd antigen has been isolated by using a K-locus specific cDNA probe. The complete nucleotide sequence of the gene reveals eight exons separated by seven introns. Transcriptionally important DNA sequences (CCAAT and TATA) precede the first exon. Comparison with other H-2 genes shows extensive homology in exons as well as in introns. Two cDNA clones encoding Kd antigens have been analysed and provide evidence for at least two expressed Kd genes in the DBA/2 mouse. Comparison of the Kd antigen sequence to three other H 2 antigens indicates that gene conversion mechanism(s) act on H-2 genes. Analyses of exon donor and acceptor sites of different H-2 genes and cDNAs show that alternative splicing sites are used by different genes. PMID- 11894936 TI - Blue light inhibits slime mold differentiation at the mRNA level. AB - The influence of blue light on protein synthesis in spherulating Physarum polycephalum microplasmodia was studied using two-dimensional protein separation techniques. The starvation-induced plasmodium-spherule transition proceeds in the dark and is accompanied by the synthesis of 20 major differentiation-specific proteins as revealed by in vivo labelling with [35S]methionine. Three of these proteins are identical with cell wall components with respect to their mol. wts. (35 K, 34 K and 14 K) and isoelectric points. Spherulation is also accompanied by the appearance of 26 prominent differentiation-specific mRNA species translatable in the rabbit reticulocyte cell-free system. Six of the proteins synthesized in vitro co-migrate on two-dimensional gels with proteins labelled in vivo, two of them being cell wall components. Blue light, which inhibits spherulation completely, inhibits also the synthesis of spherule proteins and of spherule specific mRNA activity. Only three protein components are induced by blue light, indicating that illumination does not induce a novel differentiated plasmodial state. PMID- 11894935 TI - Spatial arrangement of proteins within the small subunit of rat liver ribosomes studied by cross-linking. AB - Cross-linking of proteins within the small subunit of rat liver ribosomes by the bifunctional reagent dimethyl 4,7-dioxo-5,6-dihydroxy-3,8-diazadecanbisimidate produced numerous covalently linked protein dimers which could be separated by a combination of ion-exchange chromatography on carboxymethyl cellulose and polyacrylamide gel electrophoresis. The protein components of the dimers were identified electrophoretically after periodate cleavage of the cross-link(s). The analysis revealed 42 cross-linked dimers involving 25 different proteins. Among these, proteins S3, S4 and S20 occurred in combinations with six, eight and seven different proteins, respectively. For proteins S13, S14 and S17 five protein neighbours could be identified, while 13 of the remaining proteins were linked to three or four different protein partners. The involvement of the majority of proteins in the formation of multiple cross-linked dimers implies that a large number of protein-protein interaction sites exist within the ribosomal subunit. A preliminary model illustrating the arrangement of 16 proteins in the small ribosomal subunit is presented and discussed with respect to possible functions, especially in the event of translation initiation. PMID- 11894937 TI - Construction of novel cytochrome b genes in yeast mitochondria by subtraction or addition of introns. AB - The mitochondrial cob-box gene coding for apocytochrome b in yeast has five introns and six exons or two introns and three exons depending on the wild-type strain considered. Some intron mutations in this gene affect not only its expression but also that of another mitochondrial gene: oxi3. To understand better the function of introns in gene expression, we have constructed a series of new strains that differ only by the presence or absence of one of the five wild-type introns in the cytochrome b gene, the rest of the mitochondrial and nuclear genome remaining unchanged. All constructions result from in vivo recombination events between rho- donor and rho+ recipient mtDNA. The following genes have been constructed: [see text]. Interestingly, all the genes lead to the synthesis of cytochrome b, while only the genes having the intron bI4 allow the expression of oxi3. A nuclear gene, when mutated, can compensate for the absence of the intron bI4. PMID- 11894938 TI - Observations concerning the discontinuous DNAs of cauliflower mosaic virus. AB - The double-stranded circular DNA encapsidated within cauliflower mosaic virus (CaMV) particles contains three single-stranded discontinuities, two in one strand and one in the other, so that, upon denaturation, three linear single stranded DNAs are produced. Here we show that a fourth much smaller single stranded DNA, termed alpha1, is also present in denatured CaMV DNA preparations. The 5' extremity of alpha1 is identical to that of the alpha-strand, the strand of DNA possessing only one interruption, while its 3' extremity lies just two nucleotides downstream from a major transcription initiation site. We also show that the interrupted strand at each discontinuity sometimes has a single ribonucleotide in place of a deoxyribonucleotide at its 5' extremity. Oligoribonucleotide chains of eight and 10 residues in length have also been detected at the 5' end of one of the discontinuities. These structures are thought to be the vestiges of primers which have not been completely excised prior to encapsidation of the DNA. The possibility that synthesis of the alpha strand occurs by reverse transcription of viral RNA using initiator tRNA(met) as primer is discussed. PMID- 11894940 TI - Specific DNA binding of the cAMP receptor protein within the lac operon stabilizes double-stranded DNA in the presence of cAMP. AB - The effects of varying amounts of cAMP receptor protein (CRP) in the presence and absence of cAMP on the melting and differential melting curves of a 301-bp fragment containing the lac control region in 5 mM Na+ have been investigated. The native 301-bp fragment consists of three cooperatively melting thermalites. At 5 mM Na+, thermalite I (155 bp) has a Tm of 66.4 degrees C and the melting transitions of thermalites II (81 bp) and III (65 bp) are superimposed with a Tm of 61.9 degrees C. The specific DNA target site for CRP and the lac promotor are located within thermalite II. CRP alone exerts no specific effects on the melting of the 301-bp fragment, non-specific DNA binding of CRP resulting in a progressive stabilization of the double-stranded DNA by increasing the number of base pairs melting at a higher Tm in a non-cooperative transition. The cAMP-CRP complex, however, exerts a specific effect with a region of approximately 36 bp, comprising the specific CRP binding site and a neighbouring region of DNA, being stabilized. The appearance of this new cooperatively melting region, known as thermalite IV, is associated with a corresponding decrease in the area of thermalites II/III. The Tm of thermalite IV is 64.4 degrees C, 2.5 degrees C higher than that of thermalites II/III. With two or more cAMP-CRP complexes bound per 301-bp fragment, the stabilization also affects the remaining 110 bp now making up thermalites II/III whose Tm is increased by 1 degrees C to 62.9 degrees C. The implications of these findings for various models of the mode of action of the cAMP-CRP complex are discussed. PMID- 11894939 TI - Expression of human neuronal antigens in a mouse neuroblastoma x human dorsal root ganglion cell hybrid. AB - Clonal mouse neuroblastoma cells were fused with cells from human foetal dorsal root ganglia and several continuously-growing hybrid clones isolated. One hybrid cell line (F2.1D1) containing a number of human chromosomes, was shown to retain the ability to extend neurites in response to dibutyryl cyclic AMP and to express various antigens characteristic of human foetal dorsal root ganglion neurons. The X-chromosome-controlled 12E7 antigen, human Thy-1 and the neuron-specific F12.A2B5 antigen were identified as surface components of the hybrid cells. None of these antigens were detected in the parental neuroblastoma cell line. In addition, using a species-specific monoclonal antibody, the hybrid cells were shown to synthesize human neurofilament protein. This is the first demonstration of the continued expression of a human species- and neuron-specific gene product in a human-mouse somatic cell hybrid. PMID- 11894941 TI - Immunocytochemical localization of the gap junction 26 K protein in mouse liver plasma membranes. AB - Specific binding sites for anti-26 K antibodies directed against the liver gap junction protein (26 K) were localized by immunoelectron microscopy in gap junction plaques purified from hepatic plasma membranes. Using immunofluorescence microscopy we found discrete fluorescent spots on plasma membranes in cross sections of liver tissues after incubation with anti-26 K antibodies. This is consistent with the notion of specific binding to gap junction plaques. Quantitative binding of anti-26 K antibodies was indirectly measured by the protein A-gold technique. We found that urea/detergent-treated, purified gap junction plaques bind 30-fold more anti-26 K antibodies than preimmune serum. Anti-26 K antibodies also bind specifically to native gap junction plaques within hepatic plasma membranes although only about one fifth as efficiently as to purified plaques. Possibly the anti-26 K antibodies raised after injection of SDS denatured 26 K protein into rabbits recognize the cytoplasmic face of urea/detergent-treated plaques better than that of native plaques. Some, if not most, of the vesicular structures in preparations of purified plaques appear to be derived from split gap junction plaques and are probably sheets of gap junction hemichannels. In some vesicles the former cytoplasmic face of the hemichannels is turned outside, other vesicles have the former cell surface turned outside. The anti-26 K antibodies do not recognize any 26 K protein on the sheets of partially split gap junction plaques, on the heterogeneous vesicular structures, or on non-junctional areas of hepatic plasma membranes. These results suggest that the conformation of the 26 K protein in plaques must be different from that of the 26 K protein in earlier biosynthetic steps of plaque assembly. PMID- 11894942 TI - DNA repair dependent NAD+ metabolism is impaired in cells from patients with Fanconi's anemia. AB - In vitro cultivated fibroblasts derived either from patients with Fanconi's anemia (FA) or from healthy probands were analyzed for their DNA repair-dependent NAD+ metabolism. No difference in NAD+ pools was found. NAD+ consumption after cell damage by u.v. irradiation was, however, significantly reduced in FA cells. Several FA cell lines had a lowered ability to transfer ADP-ribose to acid precipitable material. Additionally, a decreased activity of NAD: protein ADP ribosyltransferase was found for three FA cell lines. Our data indicate, that FA is accompanied by a defective NAD+ metabolism during DNA repair. PMID- 11894944 TI - Developmental changes in the methylation of the rat albumin and alpha-fetoprotein genes. AB - We have analyzed methylation of the rat albumin and alpha-fetoprotein (AFP) genes by hydridizing labeled cDNA clones to HpaII and MspI digests of DNA from different stages of development. These CCGG-cutting enzymes distinguish 5 methylcystosine in mCCGG (sensitive to HpaII) and CmCGG (sensitive to MspI). In the liver, the albumin gene is heavily methylated at 18 days gestation and uniformly demethylated in the adult. The AFP gene is also heavily methylated at 18 days gestation, and develops demethylated regions at the 3' half of the gene in the adult. These methylation changes are not observed in other embryonic or adult tissues. We also evaluated expression of these genes by measuring their corresponding mRNAs. The albumin gene is actively transcribed in 18-day fetal liver, when it is heavily methylated, as well as in adult liver, when it is unmethylated. In contrast, the AFP gene is transcribed only in fetal liver, even though it is less methylated in adult liver. These findings suggest that specific methylation changes are associated with changes in gene expression, but that this association is not adequately described by the simple hypothesis that methylation turns genes off. PMID- 11894943 TI - A cellular protein phosphorylated by the avian sarcoma virus transforming gene product is associated with ribonucleoprotein particles. AB - In chick embryo fibroblasts transformed by Rous sarcoma virus (RSV) the tyrosine phosphorylation of a cellular protein of 34,000 daltons mol. wt. (34 kd) is greatly enhanced; this was shown to be catalyzed by the phosphotransferase activity of RSV transforming protein pp60src. We report here that in cytoplasmic extracts of both normal and transformed cells, in the presence of magnesium ions, the majority of the 34-kd protein is associated with large structures and that a fraction of 34 kd appears to be associated with ribonucleoprotein particles (RNPs). In addition, upon u.v. light cross-linking of RNA to protein in normal or transformed cells, an anti-34 kd serum immunoprecipitates RNA fragments of apparent low sequence complexity as detected by T1 fingerprint analysis. Our results indicate that the 34-kd protein may play a role in the cell at the level of RNPs. PMID- 11894945 TI - Sequences involved in the regulated expression of the human interferon-beta1 gene in recombinant SV40 DNA vectors replicating in monkey cells. AB - The human genomic EcoRI fragment of 1.83 kb containing the interferon (IFN) gene IFN-beta1 with 285 nucleotides of 5'-flanking sequences was transfected into monkey kidney CV-1 cells as part of an SV40-pML2 vector. Induction of the monkey cells to produce IFN led to a rapid accumulation of IFN-beta1 RNA whose 5' ends were identical to the IFN-beta1 mRNA of human fibroblasts. This induction occurred with all recombinants tested. Expression from the SV40 late promoter was also seen in non-induced cells. We conclude that the regulation of the IFN-beta1 gene is retained in the replicating episomal SV40 vectors with high copy number, even when the gene is being transcribed from an external promoter. When the 5' flanking sequences were deleted to leave only 40 bp before the presumed cap site of the IFN-beta1 gene, inducible formation of IFN-RNA with authentic 5' ends could still be demonstrated. However, inducibility and expression depended on the position of the deleted IFN-beta1 gene in the vector. We conclude that the sequences around the TATAA box and cap site on the IFN gene are involved in the regulation of its expression. Regulated short-term expression of the human IFN beta1 gene in SV40 vectors provides a defined system in which the structures required to maintain the regulation and the influence of known external transcription signals can be examined. PMID- 11894946 TI - The pattern of expression of chick delta-crystallin genes in lens differentiation and in trans-differentiating cultured tissues. AB - During development of the vertebrate lens, the lens epithelium undergoes a final stage of differentiation into lens fibre cells. Lens fibre cells can also be produced by trans-differentiation from certain extralenticular structures, all of which are of different developmental origin from lens, including embryonic neural retina and retinal pigmented epithelium. Delta-crystallin is the major lens protein in the chick and appears first in development; it is the major product in trans-differentiated retina of younger embryos. In both normal differentiation and trans-differentiation an increase of delta-crystallin coding RNA is detectable in the nucleus of cells prior to their terminal differentiation into lens fibres. The increase in transcription of delta-crystallin genes accompanying final differentiation of lens fibres, appears to take place slightly in advance of an increase in the capacity to process and transport this mRNA to the cytoplasm. PMID- 11894947 TI - Adenovirus cores can function as templates in in vitro DNA replication. AB - Adenovirus cores prepared by gentle disruption of virus by heating at 56 degrees C in the presence of deoxycholate were able to function as templates in an in vitro DNA replication system, allowing both initiation, indicated by the formation of terminal protein-dCMP complex, and elongation of > 300 nucleotides. Using both cores and DNA-protein complexes as templates, it was also demonstrated that novobiocin, an inhibitor of DNA gyrase, inhibited in vitro DNA replication by preventing formation of the initiation complex. PMID- 11894948 TI - Bacteriophage Mu DNA circularizes following infection of Escherichia coli. AB - Mu DNA, isolated from infected cells or minicells, has been shown to be held by proteins in twisted and open circular forms. Circularization does not require protein synthesis in the infected cells. A 64,000-dalton polypeptide is injected into the infected cell with Mu DNA and co-sediments with Mu DNA through sucrose gradients. Circularization of the infecting Mu DNA does not require removal of the Escherichia coli DNA sequences which are attached to both ends of the Mu genome in the viral particle. PMID- 11894949 TI - Altered maturation of sequences at the 3' terminus of 5S gene transcripts in a Saccharomyces cerevisiae mutant that lacks a RNA processing endonuclease. AB - Sequences at the immediate 3' terminus of several eukaryotic primary transcripts, synthesised just before the termination of transcription, are often lost during RNA processing. The rna82.1 mutation in Saccharomyces cerevisiae appears to result in a deficiency of the endonuclease that removes such sequences from certain yeast transcripts. Some small RNAs of rna82.1 cells are a few nucleotides longer than their counterparts in wild-type S. cerevisiae. The 5S rRNAs made during very short pulse-labellings of the mutant have, relative to the mature 121 nucleotide 5S RNA of wild-type cells, an additional 7, 11 or 13 nucleotides at their 3' terminus. These 5S forms reveal sites upon 5S genes where transcription probably terminates in vivo. The extra nucleotides upon 5S RNAs in rna82.1 cells are lost very slowly by sequential removal from the 3' terminus. Through this 3' 5' exonuclease action the total 5S RNA of the mutant possesses several 3' terminal sequences yet is mostly only 0-3 nucleotides longer than in wild-type S. cerevisiae. Just one or two of these 3'-terminal sequences serve as a substrate in vivo for a poly(A) polymerase since a small proportion of rna82.1 5S RNAs terminate in the sequence: CAAUCUUU(A)n. PMID- 11894950 TI - A monoclonal antibody which recognises each of the nuclear lamin polypeptides in mammalian cells. AB - A monoclonal IgM has been characterised which recognises the nuclear lamins in all mammalian cells tested. In immunoblotting experiments using both one- and two dimensional gels it recognises lamins A, B and C. The common antigenic determinant lies on a proteolytic fragment of 46,000 daltons which can be generated from each lamin polypeptide by treatment with chymotrypsin. In immunofluorescence experiments on whole cells and thin frozen sections, the antibody labelled only the nuclear envelope and not the nuclear interior. During mitosis, labelling was found dispersed throughout the cell cytoplasm. By immunoelectron microscopy using the antibody and protein A-gold, only the nucleoplasmic side of the nuclear envelope (the nuclear lamina) was labelled, but there was no labelling of the nuclear pores. PMID- 11894951 TI - Phosphorylation and actin activation of brain myosin. AB - A method is described for obtaining brain myosin that shows significant actin activation, after phosphorylation with chicken gizzard myosin light chain kinase. Myosin with this activity could be obtained only via the initial purification of brain actomyosin. The latter complex, isolated by a method similar to that used for smooth muscle, contained actin, myosin, tropomyosin of the non-muscle type and another actin-binding protein of approximately 100,000 daltons. From the presence of a specific myosin light chain kinase and phosphatase in brain tissue it is suggested that the regulation of actin-myosin interaction operates via phosphorylation and dephosphorylation of myosin. PMID- 11894952 TI - Degrees of relatedness of T-even type E. coli phages using different or the same receptors and topology of serologically cross-reacting sites. AB - The relatedness of a series of T-even like phages which use the Escherichia coli outer membrane protein OmpA as a receptor, and the classical phages T2, T4 and T6 has been investigated. Immunoelectron microscopy and the pattern of phage resistance in bacterial mutants revealed that: (i) phages of this morphology do not necessarily cross-react serologically; (ii) phages using different receptors may bind heterologous IgG everywhere except to the tip (comprising approximately 10% of one fiber polypeptide) of the long tail fibers; (iii) cross-reacting OmpA specific phages may bind heterologous IgG only to the tip of these fibers: (iv) OmpA-specific phages not cross-reacting at the tip of the tail fibers use different receptor sites on the protein. Absence of cross-reactivity appears to reflect high degrees of dissimilarity. A DNA probe consisting of genes encoding the two most distal tail fiber proteins of T4 detected homologies only in DNA from phages serologically cross-reacting at this fiber. Even under conditions of low stringency, allowing the formation of stable hybrids with almost 30% base mismatch, no such homologies could be found in serologically unrelated phages. Thus, in the collection of phages examined, there are sets of very similar and very dissimilar tail fiber genes and even of such gene segments. PMID- 11894953 TI - High affinity binding of alpha-bungarotoxin to the purified alpha-subunit and to its 27,000-dalton proteolytic peptide from Torpedo marmorata acetylcholine receptor. Requirement for sodium dodecyl sulfate. AB - Intact nicotinic acetylcholine receptor (AChR) tightly binds alpha-bungarotoxin. The two toxin-binding sites are presumed to be on the two alpha-subunits, either on or near the ACh-binding sites. Isolated alpha-subunits have been found to maintain weak binding to alpha-bungarotoxin (KD approximately 0.2 microM). We describe here conditions under which the alpha-subunit and a 27,000-dalton proteolytic peptide bound alpha-bungarotoxin with high affinity. The four subunits of Torpedo marmorata AChR, as well as several proteolytic peptides of the alpha-subunit, were first purified by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. We found that the purified alpha-subunit (but not the beta-, gamma- or delta-subunits) and its 27,000-dalton peptide specifically bound 125I-labeled alpha-bungarotoxin with KD approximately 3 and 6 nM, i.e., about two orders of magnitude lower than the intact AChR. Nearly 100% of the sites were recovered. The recovery of this high affinity binding required the presence of SDS (approximately 0.02%) but non-denaturing detergents had a strongly inhibitory effect. Unlabeled alpha-toxins competed with labeled alpha bungarotoxin, alpha-bungarotoxin being more effective than all the other toxins tested. Decamethonium and hexamethonium competed efficiently with alpha bungarotoxin binding but carbamylcholine had only a weak effect. The main immunogenic region of the AChR was only partially preserved since conformation dependent monoclonal antibodies to this region bound the alpha subunit-toxin complexes, but much less efficiently than the intact AChR. We conclude that SDS can be advantageous to the recovery of high toxin binding to the alpha subunit which still has not completely recovered its native conformation. PMID- 11894954 TI - Complete sequence of an HLA-dR beta chain deduced from a cDNA clone and identification of multiple non-allelic DR beta chain genes. AB - At least three polymorphic class II antigens are encoded in the human major histocompatibility complex (HLA): DR, DC and SB. cDNA clones encoding beta chains of HLA-DR antigen, derived from mRNA of a heterozygous B-cell line, were isolated and could be divided into four subsets, clearly distinct from cDNA clones encoding DC beta chains. Therefore, at least two non-allelic DR beta chain genes exist. The complete sequence of one of the DR beta chain cDNA clones is presented. It defines a putative signal sequence, two extracellular domains, a trans-membrane region and a cytoplasmic tail. Comparison with a DC beta chain cDNA clone revealed a homology of 70% between the two beta chains and that the two genes diverged under relatively little selective pressure. A set of amino acids conserved in immunoglobulin molecules was found to be identical in both DR and DC beta chains. Comparison of the DR beta chain sequence with the amino acid sequence of another DR beta chain revealed a homology of 87% and that most differences are single amino acid substitutions. Allelic polymorphism in DR beta chains has probably not arisen by changes in long blocks of sequence. PMID- 11894955 TI - Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster. AB - The distribution patterns of nonhistone chromosomal proteins (NHCP) associated with pulse-labeled RNA were determined by indirect immunofluorescence on salivary gland chromosomes of Drosophila melanogaster using monoclonal antibodies. By staining for two different antigens simultaneously, using antibodies tagged with different fluorescent probes, it became possible to position RNA-associated antigens as well as RNA polymerase B in relation to each other. Three separate staining patterns could be observed with anti-NHCP antibodies, none of which showed a pattern which was identical with that of RNA polymerase B. Furthermore, no correlation with the synthesis of the primary trancript, as monitored by the RNA polymerase B content of chromosomal sites, could be found by following the fluorescence patterns during inactivation of intermolt puffs or activation of early ecdysone-induced puffs. Finally, no strict correlation was observed between puffing activity and the accumulation of a certain antigen in these selected chromosomal sites. PMID- 11894956 TI - The conserved part of the T-region in Ti-plasmids expresses four proteins in bacteria. AB - The T-region of Ti-plasmids expresses four proteins (mol. wts. 74,000, 49,000, 28,000 and 27,000) in Escherichia coli minicells. Promoter activities are determined by sequences within the T-region, and the protein-coding regions map in that part of the T-region which is highly conserved in octopine and nopaline plasmids and which is responsible for shoot and root inhibition when expressed in plant cells. Three of the regions expressed in bacteria correlate with three regions which are transcribed in transformed plant cells; the fourth protein coding region has no corresponding transcript in plants. At least three of the proteins synthesized in E. coli minicells are also expressed in cell-free systems prepared from E. coli and from Agrobacterium tumefaciens; the fourth protein (mol. wt. 49,000) is poorly expressed in both cell-free extracts. The possibility is discussed that the same genes are expressed in Agrobacteria and in transformed plant cells and that in both cases the gene products mediate growth regulatory effects to non-transformed plant cells. PMID- 11894957 TI - Intergeneric transfer and exchange recombination of restriction fragments cloned in pBR322: a novel strategy for the reversed genetics of the Ti plasmids of Agrobacterium tumefaciens. AB - Transmission of ColE1/pMB1-derived plasmids, such as pBR322, from Escherichia coli donor strains was shown to be an efficient way to introduce these plasmids into Agrobacterium. This was accomplished by using E. coli carrying the helper plasmids pGJ28 and R64drd11 which provide the ColE1 mob functions and tra functions, respectively. For example, the broad host-range replication plasmid, pGV1150, a co-integrate plasmid between pBR322 and the W-type mini-Sa plasmid, pGV1106, was transmitted from E. coli to A. tumefaciens with a transfer frequency of 4.5 x 10(-3). As pBR322 clones containing pTiC58 fragments were unable to replicate in Agrobacterium, these clones were found in Agrobacterium only if the acceptor carried a Ti plasmid, thus allowing a co-integration of the pBR322 clones with the Ti plasmid by homology recombination. These observations were used to develop an efficient method for site-specific mutagenesis of the Ti plasmids. pTiC58 fragnents, cloned in pBR322, were mutagenized in vitro and transformed into E. coli. The mutant clones were transmitted from an E. coli donor strain containing pGJ28 and R64drd11 to an Agrobacterium containing a target Ti plasmid. Selecting for stable transfer of the mutant clone utilizing its antibiotic resistance marker(s) gave exconjugants that already contained a co integrate plasmid between the mutant clone and the Ti plasmid. A second recombination can dissociate the co-integrate plasmid into the desired mutant Ti plasmid and a non-replicating plasmid formed by the vector plasmid pBR322 and the target Ti fragment. These second recombinants lose the second plasmid and they are identified by screening for the appropriate marker combination. PMID- 11894958 TI - Identification of sequences involved in the polyadenylation of higher plant nuclear transcripts using Agrobacterium T-DNA genes as models. AB - Sequences in the 3'-untranslated region of two different octopine T-DNA genes were analyzed with regard to their significance in polyadenylation. Poly(A) addition sites were localized precisely by S1 nuclease mapping with T-DNA-derived mRNAs isolated from tobacco. The gene encoding transcript 7' contains two AATAAA hexanucleotides, respectively 119 bp and 170 bp downstream of the TAA stop codon. A single poly(A) site was mapped 24-25 bp downstream of the first AATAAA. Further, we show that a mutant octopine synthase gene, which has lost part of its 3'-untranslated region by deletion, is still active. This mutant gene terminates 19 bp upstream from the major wild-type polyadenylation site. The deletion also removes the AATAAT signal preceding this site. The mutant octopine synthase gene contains a minimum of four different poly(A) sites. The most prominent of these sites is identical to the minor poly(A) site of the wild-type gene, and is preceded by a sequence AATGAATATA. Three other sites are located within the adjacent plant DNA, giving rise to hybrid T-DNA/plant DNA transcripts. The two most distal sites are probably dependent on a motif AATAAATAAA, found 29 bp away from the T-DNA/plant DNA junction. PMID- 11894959 TI - The mitochondnal genome of Aspergillus nidulans contains reading frames homologous to the human URFs 1 and 4. AB - A 2830-bp segment of the mitochondrial genome of the fungus Aspergillus nidulans was sequenced and shown to contain two unidentified reading frames (URFs). These reading frames are 352 and 488 codons in length, and would specify unmodified proteins of mol. wts. 39,000 and 54,000, respectively. The derived amino acid sequences indicate that these genes are equivalent to the human mitochondrial URFs 1 and 4, with 39% amino acid homology for URF1 and 26% for URF4. Both URFs were shown by secondary structure predictions to code for predominantly beta sheeted proteins with strong structural conservation between the fungal and human homologues. Counterparts of mammalian URFs have not previously been identified in non-mammalian genomes, and the discovery that A. nidulans possesses reading frames so closely homologous with URF1 and URF4 shows that these genes are of general functional importance in the mitochondria of diverse species. PMID- 11894960 TI - Multiplicity of constant kappa light chain genes in the rabbit genome: a b4b4 homozygous rabbit contains a kappa-bas gene. AB - We have constructed a genomic library of homozygous b4b4 rabbit DNA in the pJB8 cosmid vector. Clones containing Ckappa-like sequences were screened with a b4 cDNA probe and were characterized by restriction mapping. One of the clones contained a Ckappa sequence different from the b4 allotype normally expressed by the animal. We report here the nucleotide sequence of this gene and show that it probably corresponds to a kappa-bas form of the Basilea allotype. It appears to be a structurally complete gene without any stop codons within the coding region and containing the dinucleotide AG as a splice site acceptor for the J-C junction, just 5' of the coding block. Comparison with the b4 cDNA nucleotide sequence shows a separate evolution of the Ckappa-coding and 3'-untranslated sequences, since the 3'-untranslated regions are more conserved than the coding regions. Genomic blot analysis would suggest that the kappa-bas gene is isotypic in the domestic rabbit population, since it lies within a genomic EcoRI or PstI restriction fragment, which was shown to be common to all homozygous b4, b5, b6 and b9 rabbit DNAs. PMID- 11894961 TI - Patterns of major divergence between the internal transcribed spacers of ribosomal DNA in Xenopus borealis and Xenopus laevis, and of minimal divergence within ribosomal coding regions. AB - We have determined the nucleotide sequences of the two internal transcribed spacers, the adjacent ribosomal coding sequences and the boundary between the external transcribed spacer and the 18S coding sequence in a cloned ribosomal transcription unit from Xenopus borealis. The transcribed spacers differ very extensively from those of X. laevis. Nevertheless, embedded in the internal transcribed spacers are several short sequence elements which are identical between the two species. These conserved elements are laterally displaced by substantial distances in the X. borealis sequence with respect to that of X. laevis. These relative displacements imply that insertions and deletions have played a major role in transcribed spacer divergence in Xenopus. This in turn implies that large regions of the transcribed spacers do not play a sequence specific role in ribosome maturation. In contrast, the sequenced parts of the ribosomal coding regions, which encompass 670 nucleotides, differ at only three points from the corresponding sequences in X. laevis, each by a single substitution. These substitutions are readily accommodated by current models for rRNA higher order structure. PMID- 11894962 TI - The structure of an unusual leghemoglobin gene from soybean. AB - A clone containing an unusual leghemoglobin (Lb) gene was isolated from a soybean DNA library present in Charon 4A phage. DNA sequence analysis revealed that the isolated Lb gene has three intervening sequences (IVS-1, IVS-2 and IVS-3) located in the same positions as those found in other Lb genes. Due to a large increase of IVS-2 and IVS-3, the isolated Lb gene is about twice the size of a normal Lb gene. The coding sequence derived from the DNA sequence corresponds to no known soybean Lb and attempts to find a corresponding mRNA failed. In addition, the 5' flanking sequence of the Lb gene is mutated in two regions which seem to be important for transcription. It is, therefore, tentatively suggested that the isolated Lb gene is non-functional, and consequently is an Lb pseudogene. PMID- 11894963 TI - The DNA sequence of the H-2kb gene: evidence for gene conversion as a mechanism for the generation of polymorphism in histocompatibilty antigens. AB - We have determined the DNA sequence of the H-2Kb gene of the C57B1/10 mouse. Comparison of this sequence with that of the allelic H-2Kd shows surprisingly that the exons have accumulated more mutations than their introns. Moreover, many of these changes in the exons are clustered in short regions or hot spots. Additional comparison of these sequences with the H-2Ld and H-2Db sequences shows that, in several cases, the altered sequence generated at the hot spot is identical to the corresponding region of a non-allelic H-2 gene. The clustered changes are responsible for 60% of the amino acid differences between the H-2Kb and H-2Kd genes and suggest that micro-gene conversion events occurring within the exons and involving only tens of nucleotides are an important mechanism for the generation of polymorphic differences between natural H-2 alleles. PMID- 11894966 TI - New therapeutic approaches to type 1 diabetes: from prevention to cellular or gene therapies. PMID- 11894967 TI - The role of chemotherapy in the nonsurgical management of malignant neuroendocrine tumours. PMID- 11894965 TI - Characterization of an integral membrane glycoprotein associated with the microfilaments of pig intestinal microvilli. AB - An integral membrane glycoprotein of pig intestinal microvilli which exists in two polypeptide forms [mol. wt. 140 K and 200 K as measured by SDS-polyacrylamide gel electrophoresis (SDS-PAGE)] was purified to homogeneity and characterized. The 200-K form is probably a precursor of the 140-K species. We have localized the glycoprotein by electron microscope immunochemistry using specific antibodies and determined its topological organization with respect to the membrane bilayer. Triton X-100 treatments which solubilize most other microvillar membrane glycoproteins from purified, closed, right-side out vesicles do not efficiently extract this protein. The protein can be partially solubilized from the detergent insoluble residue, either by treatment with proteases (trypsin or papain) or by exposure to low ionic strength buffer in the presence of chelating agents and detergents. Once solubilized by papain or trypsin, the protein co-migrates on SDS PAGE with the protein obtained by low ionic strength extraction. However, the form of the protein released by papain does not bind detergents and exhibits hydrophilic properties. Our observations are consistent with the 140-K protein having a small hydrophobic domain that anchors it to the microvillar membrane. The 140-K glycoprotein binds in vitro to a 110-K protein of the core cytoskeleton residue. These observations suggest that the 140-K glycoprotein may be a transmembrane protein which may in vivo provide attachment sites for direct or indirect association with polypeptides of the microvillus cytoskeleton. PMID- 11894964 TI - The origin of replication, oriC, and the dnaA protein are dispensable in stable DNA replication (sdrA) mutants of Escherichia coli K-12. AB - The sdrA224 mutants of Escherichia coli K-12, capable of continued DNA replication in the absence of protein synthesis (stable DNA replication), tolerate inactivation of the dnaA gene by insertion of transposon Tn10. Furthermore, oriC, the origin of E. coli chromosome replication, can be deleted from the chromosome of sdrA mutants without loss of viability. The results suggest the presence of a second, normally repressed, initiation system for chromosome replication alternative to the 'normal' dnaA+ oriC+-dependent initiation mechanism. PMID- 11894968 TI - Ghrelin, not just another growth hormone secretagogue. PMID- 11894969 TI - Corticotrophin-releasing hormone and parturition. PMID- 11894972 TI - Is there an increase in second brain tumours after surgery and irradiation for a pituitary tumour? AB - OBJECTIVE: To assess the incidence of second brain tumours in patients operated and irradiated for pituitary tumours. DESIGN AND PATIENTS: The study base consisted of a consecutive series of 325 patients operated and irradiated for pituitary tumours, excluding patients with acromegaly and Cushing's disease. Comparison was made with the general population from the same catchment area as the patients. The follow-up period started in 1958 and on an individual basis patients were followed from the onset of postoperative irradiation until December 1995, or until date of death, emigration or a second brain tumour diagnosis, whichever occurred first. RESULTS: Three brain tumours (two astrocytomas and one meningioma) were observed, compared with 1-13 expected (standardized incidence ratios (SIR) 2.7; 95% confidence interval (CI) 0.6-7.8). CONCLUSION: The present study gives no firm support for an increased incidence of a second brain tumour in patients operated and irradiated for pituitary tumours. A crude meta-analysis of the present and previously published cohort studies of patients with irradiated pituitary tumours gives an SIR of 6.1 (95% CI 3.16-10.69). Thus, the results of the meta-analysis are in favour of an increased risk for second brain tumours. A genetic trait that predisposes to both pituitary tumours and brain tumours is an alternative causal factor. There is no definite proof that cranial irradiation per se is the causal factor. This question cannot be fully answered until sufficient cohort studies of nonirradiated pituitary tumour patients have been carried out. PMID- 11894971 TI - Relationships between the taqI polymorphism of the dopamine D2 receptor and blood pressure in hyperglycaemic and normoglycaemic Chinese subjects. AB - BACKGROUND: We have previously reported an association of the A2 allele of the dopamine D2 receptor (DRD2) TaqI polymorphism with increased blood pressure in normoglycaemic Chinese subjects, but conversely possibly with decreased indices of obesity. Hypertension is also a common feature of patients with type 2 diabetes, with up to 50% being hypertensive. OBJECTIVE: To compare the relationship between the DRD2 TaqI polymorphism, blood pressure and obesity in Chinese patients with and without fasting hyperglycaemia. METHOD: The DRD2 TaqI polymorphism was determined by PCR-RFLP in 519 normoglycaemic and 471 hyperglycaemic Chinese subjects, of whom 53.2 and 48.8% were hypertensive, respectively. RESULTS: In the normoglycaemic subjects there was a significant increase in mean arterial pressure (P= 0.041) with increasing proportions of the A2 allele, 95 +/- 16, 96 +/- 17 and 100 +/- 17 mmHg for the A1A1, A1A2 and A2A2 genotypes, respectively. However, the relationship was not observed in the subjects with fasting hyperglycaemia either in the total group or in the subgroup who were not receiving blood pressure-lowering medication (n = 383, 97 +/- 15, 98 +/- 14 and 97 +/- 15 mmHg, respectively). When the whole group was divided into those subjects obese by either body mass index or waist-to-hip ratio (n = 484) and those subjects not obese by both these criteria (n= 506), the A1 allele (49.2 vs. 43.8%, P = 0.02) and A1 allele containing genotypes (P = 0.03) were more frequent in the obese subjects. Similar relationships were seen in the normoglycaemic and hyperglycaemic groups separately, although these did not reach significance. CONCLUSIONS: In the normoglycaemic subjects, the A2 allele was associated with increased blood pressure and possibly lower indices of obesity, but in the hyperglycaemic subjects only the possible association with obesity was noted. PMID- 11894970 TI - Cytotoxic T lymphocyte antigen 4 gene polymorphism confers susceptibility to type 1 diabetes in Japanese children: analysis of association with HLA genotypes and autoantibodies. AB - OBJECTIVE: Although the polymorphisms of the cytotoxic T lymphocyte antigen 4 (CTLA4) gene have been shown to be associated with Type 1 diabetes in Caucasians, some conflicting results have been reported among subjects of different ethnic backgrounds. We examined a CTLA4 polymorphism and its relationship to human leucocyte antigen (HLA) genotypes and autoantibodies for glutamic acid decarboxylase 65 (GAD65) and IA-2 in Japanese children with Type 1 diabetes. SUBJECTS AND MEASUREMENTS: The study group consisted of 125 childhood-onset Japanese subjects (50 males, 75 females) with Type 1 diabetes. The CTLA4 A/G polymorphism at position 49 was analysed using a PCR-restriction fragment length polymorphism (PCR-RFLP) method. The HLA-DRB1 and DQB1 genotypes were defined by DNA analysis using PCR-sequence-specific oligonucleotide (PCR-SSO) probes. The GAD65 autoantibody (GAD65Ab) and IA-2 autoantibody (IA-2Ab) titres were measured using radioimmunoassay. RESULTS: The distribution of genotype frequencies differs between subjects with Type 1 diabetes (GG: 46%, AG: 50%, AA: 5%) and controls (GG: 39%, AG: 44%, AA: 17%) (P < 0.01). The frequency of the G allele is higher in the diabetes group than in the controls (P < 0.05). When the subjects were subdivided according to HLA genotype, the two major HLA high-risk groups, with DR9-DQ9 and DR4-DQ4, that are unique to Japanese populations showed no difference in their CTLA4 polymorphism frequencies. Although no association between the CTLA4 polymorphism and the prevalence of GAD65Ab was found, CTLA4 GG subjects that had been newly diagnosed (< 9 months) had significantly higher levels of autoantibodies than AG subjects (P < 0.01). The prevalence and titres of IA-2Ab were not associated with the CTLA4 polymorphism. CONCLUSIONS: The CTLA4 gene might confer a susceptibility to childhood-onset Type 1 diabetes in the Japanese population. The association between this CTLA4 polymorphism and the HLA genotype was similar for both major groups with HLA high-risk alleles. CTLA4 might contribute to the humoral immune response to GAD in newly diagnosed subjects. PMID- 11894973 TI - Currently used growth-promoting treatment of children results in normal bone mass and density. A prospective trial of discontinuing growth hormone treatment in adolescents. AB - BACKGROUND AND AIMS: The need for continued GH replacement in patients with childhood-onset GH deficiency (GHD) into adulthood has been recognized. The consequences of discontinuing GH treatment on bone mineralization in adolescent patients with GHD and short stature were examined over a period of 2 years. PATIENTS: Forty adolescents (aged 16-21 years) treated with GH for more than 3 years and 16 closely matched healthy controls were studied. After a baseline visit, GH treatment was discontinued. The patients were then re-examined with the same protocol after 1 and 2 years. Twenty-one patients had continuing severe GHD into adulthood, while 19 patients were regarded as having sufficient endogenous GH secretion (GHS). RESULTS: At baseline, there were no differences between the groups in total bone mineral content (BMC) or bone mineral density (BMD). After 2 years without GH treatment, BMC increased similarly in the GHD and GHS groups. BMC of the lumbar spine (L2-L4) increased only in the GHD group. Lumbar spine BMD increased in the GHD and the GHS groups. No changes were observed in the femoral neck region. Biochemical measurements showed that carboxy-terminal cross-linked telopeptide of type I collagen (ICTP) and bone specific alkaline phosphates (ALP) were higher in the GHD and GHS groups at baseline compared with controls. Osteocalcin, carboxy-terminal propeptide of type I procollagen (PICP), ICTP and ALP decreased during the 2 years off treatment in both the GHD and GHS groups. PICP was also lower after 2 years in the GHD group compared with both the GHS group and controls. CONCLUSIONS: After discontinuation of GH therapy in adolescents at or near final height, there was a continued increase in BMC and BMD both for adolescents with growth hormone deficiency and for those classified as growth hormone sufficient. These groups did not differ from controls at baseline or after 2 years. In the growth hormone deficiency group, biochemical markers for bone formation decreased to levels below those in the growth hormone sufficient and healthy control groups. Although the number of patients and controls in this study were small, the results indicate that the present treatment of Swedish GH-deficient children to final height results in normal BMD. PMID- 11894974 TI - Adrenocorticotropin stimulation tests in patients with hypothalamic-pituitary disease: low dose, standard high dose and 8-h infusion tests. AB - OBJECTIVES: Low doses of ACTH [1-24] (0.1, 0.5 and 1.0 microg per 1.73 m2) may provide a more physiological level of adrenal stimulation than the standard 250 microg test, but not all studies have concluded that the 1.0 microg is a more sensitive screening test for central hypoadrenalism. Eight-hour infusions of high dose ACTH [1-24] have also been suggested as a means of assessing the adrenals' capacity for sustained cortisol secretion. In this study, we compared the diagnostic accuracy of three low dose ACTH tests (LDTs) and the 8-h infusion with the standard 250 microg test (HDT) and the insulin hypoglycaemia test (IHT) in patients with hypothalamic-pituitary disease. SUBJECTS AND DESIGN: Three groups of subjects were studied. A healthy control group (group 1, n = 9) and 33 patients with known hypothalamic or pituitary disease who were divided into group 2 (n = 12, underwent IHT) and group 3 (n = 21, IHT contraindicated). Six different tests were performed: a standard IHT (0.15 U/kg soluble insulin); a 60 minute 250 microg HDT; three different LDTs using 0.1 microg, 0.5 microg and 1.0 microg (all per 1.73 m2); and an 8-h infusion test (250 microg ACTH [1-24] at a constant rate over 8 h). RESULTS: Nine out of the 12 patients in group 2 failed the IHT. Three out of 12 patients from group 2 who clearly passed the IHT, also passed all the ACTH [1-24] stimulation tests. Seven of the 9 patients who failed the IHT, failed by a clear margin (peak cortisol < 85% of the lowest normal). Two of the 7 also failed all the ACTH [1-24] tests. Five of the 7 patients had discordant results, four passed the 0.1 LDT, one (out of four) passed the 0.5 LDT, none (out of three) passed the 1.0 LDT, two passed the HDT and three passed the 8-h test. Two patients were regarded as borderline fails in the IHT. Both passed the ACTH [1-24] tests, although one was a borderline pass in the 8-h test. Only five out of the 21 patients in group 3 showed discordance between the HDT and the LDTs. One patient passed the HDT and failed the 0.1 LDT, four patients failed the HDT but passed some of the different LDTS. CONCLUSIONS: We conclude that in the diagnosis of central hypoadrenalism, ACTH [1-24] stimulation tests may give misleading results compared to the IHT. The use of low bolus doses of ACTH [1-24] (1.0, 0.5 or 0.1 microg) or a high dose prolonged infusion does not greatly improve the sensitivity of ACTH [1-24] testing. Dynamic tests that provide a central stimulus remain preferable in the assessment of patients with suspected ACTH deficiency. PMID- 11894975 TI - Plasma homocysteine is not a major risk factor for vascular disease in growth hormone deficient adults. AB - OBJECTIVE: Several cardiovascular risk factors have been investigated in patients with adult growth hormone deficiency (GHD) to explain the observed increase in vascular mortality. Plasma homocysteine concentration has been identified recently as an independent risk factor for atherosclerosis. We wished to determine whether plasma homocysteine contributes to cardiovascular risk in adult GHD. METHOD: Plasma homocysteine was measured by fluorescence polarization immunoassay in 45 GH-deficient adults on stable conventional hormone replacement (25M, 20F), age range 23-76 years, and compared with 55 matched controls (30M, 25F), age range 21-77 years. All subjects were free from clinical hypertension, diabetes, ischaemic heart disease and peripheral vascular disease. Blood pressure, body mass index and waist hip ratio were recorded. Serum creatinine and fasting lipids were measured. Serum vitamin B12 and folate levels, important cofactors in the homocysteine metabolic pathways, were also measured. RESULTS: Homocysteine levels were not different in patients and controls (9.75 [7.8-11.6] micromol/l vs. 9.65 [8.3-11.5] micromol/l, respectively, P = 0.88). Serum vitamin B12 was also not different (320.5 [262.0-427.5] pmol/l vs. 313.5 [277.0-460.5] pmol/l, respectively, P = 0.77). Serum folate levels were significantly lower in the patient group (7.05 [5.12-8.27] ng/ml vs. 7.80 [6.52-10.60] ng/ml, respectively, P = 0.03). When separated by gender, in males folate was not significantly different between patients and controls 7.05 [5.17-9.19] vs. 7.65 [6.15-10.22], P = 0.264, whereas in females, folate was significantly lower in patients at 7.05 [4.57-7.75] compared to controls at 8.4 [6.60-12.20], P = 0.01. CONCLUSION: Plasma homocysteine levels are not significantly elevated in GH deficient adults and are unlikely to be a major risk factor for vascular disease in these individuals. PMID- 11894976 TI - Alcohol ingestion decreases both diurnal and nocturnal secretion of leptin in healthy individuals. AB - OBJECTIVE: Neuropeptide Y (NPY) stimulates appetite and increases food intake. Leptin inhibits NPY. It is not known whether alcohol influences any of these factors, but it has been suggested that alcohol stimulates appetite in man. The primary objective of this investigation was to determine whether ingestion of ethanol inhibits leptin secretion in normal subjects. SUBJECTS AND DESIGN: Fourteen healthy, non-obese subjects of both sexes (7 F/7 M) were included. They were divided into two groups (I and II; 8/6). All were investigated on two occasions. On one occasion alcohol was ingested, and on the other drinking water was given. The experiments took place in random order, one week apart. In group I two experiments (A = alcohol; B = water) were performed during the day. In group II the experiments were carried out during the night (C = alcohol; D = water). Each alcoholic drink contained 0.45 g ethanol/kg. The drinks were given at 08.00, 09.30 and 11.00 hours in experiments A and B, and at 18.00, 20.00 and 22.00 hours in experiments C and D. Venous blood samples were collected before, during and after the drinks over periods of 6 h in group I and 14 h in group II. MEASUREMENTS: Serum concentrations of leptin, insulin-like growth factor 1 (IGF 1), IGF binding protein 1 (IGFBP-1), insulin, cortisol, testosterone, ethanol and plasma glucose were determined. RESULTS Group I serum leptin levels declined during the day in both men and women regardless of whether alcohol or water had been administered in the morning. Since leptin levels in general were markedly higher in women than in men, all leptin changes after water/alcohol were transformed to percentage changes to make them comparable between sexes. When the percentage leptin decline over a 6-h period (08.00-24.00 hours) was expressed by a decremental area under curve (AUC08-14), it became evident that alcohol inhibited leptin secretion, inasmuch as the leptin decremental area, obtained after alcohol, was significantly larger than the one obtained after water (124 +/ 17 vs. 57 +/- 8; P < 0.01). Similar insulin and glucose levels were obtained after alcohol and water. Group II serum leptin levels increased after both alcohol and water during the initial part of the night (18.00-20.00 hours). In this period alcohol inhibited the secretion of leptin as shown by the leptin incremental area (AUC18-24) which was 53 +/- 18 after alcohol and 113 +/- 15 after water (P < 0.01). As the ethanol concentration in serum began to fall, its inhibitory effect on leptin gradually disappeared, and when leptin AUCs representing the entire night were determined after alcohol and water, they were not significantly different. Similar insulin, glucose, testosterone, IGF-1 and cortisol levels were found after alcohol and water. The IGFBP-1 level increased, but not significantly so until 6 h after commencing the alcohol ingestion. CONCLUSION: Ingestion of moderate amounts of alcohol has an inhibitory effect on leptin secretion in normal subjects. The effect may be direct rather than indirect, since several factors known to affect leptin are not influenced by alcohol. It is tempting to speculate that alcohol might serve as an appetizer by decreasing leptin secretion, but additional studies are necessary to prove that hypothesis since previous studies have shown that leptin has a long-term rather than an acute effect on hunger. PMID- 11894977 TI - The CAG repeat polymorphism in the androgen receptor gene affects bone density and bone metabolism in healthy males. AB - OBJECTIVE: Bone metabolism and bone density (BD) are influenced by sex hormones. Testosterone (T) action is exerted through the androgen receptor (AR). We investigated the potential impact of the CAG repeat (CAGR) polymorphism within the AR gene on BD and bone metabolism in healthy younger males. PATIENTS AND MEASUREMENTS: The number of CAGRs in 110 healthy men aged 20-50 years was determined by sequence analysis. We assessed BD by the radiation-free method of quantitative ultrasound (QUS) of the phalanges. Serum levels of bone-specific alkaline phosphatase (BAP) and urine secretion of free deoxypyridinoline (DPD, corrected for creatinine), serum levels of sex hormones, body fat content and lifestyle factors were determined. RESULTS: In stepwise multiple regression models controlling for age, body fat content and lifestyle factors, the number of CAGRs was an independent negative predictor of BD (partial r = - 0.286, P = 0.001), whereas it was positively associated with markers of bone turnover (for BAP: partial r = 0.32, P= 0.001; for DPD: partial r = 0-241, P = 0.013). Levels of free T and oestradiol showed an independent and positive association with BD; age contributed significantly to lower BD. Age and free T were negatively associated with markers of bone turnover, whereas oestradiol showed a positive correlation with BAP and DPD. ANOVA in groups according to age and the CAGR length suggested an increased age-dependent bone loss in subjects with a CAGR length of 22-31 compared with 14-21 CAGRs (overall P < 0.01). CONCLUSIONS: A high number of CAG repeats within the androgen receptor gene attenuates testosterone effects on bone density and bone metabolism. This seems to be associated with accelerated age-dependent bone loss. PMID- 11894979 TI - Polycystic ovary syndrome after precocious pubarche: ontogeny of the low birthweight effect. AB - OBJECTIVE: Young girls with precocious pubarche (PP) are at increased risk of developing polycystic ovary syndrome (PCOS), including hyperinsulinism, dyslipidaemia and ovarian hyperandrogenism, particularly if PP itself was preceded by a low birthweight. Resistance to insulin is thought to be a key factor in the pathogenesis of this sequence. We aimed to elucidate the peripubertal ontogeny of the low birthweight effect on hyperinsulinism, dyslipidaemia and ovarian dysfunction after PP. PATIENTS AND DESIGN: We obtained fully longitudinal data from 51 girls with a history of PP and compared normal birthweight (n = 26) with low-birthweight (n = 25) girls (birthweight SD score 0.0 +/- 0-2 vs. - 2.4 +/- 0.2) for measurements obtained at diagnosis of PP (mean age 7.0 years), in early puberty (10.4 years) and after menarche (14.3 years). MEASUREMENTS: Fasting serum lipids and lipoproteins, together with insulin responses to an oral glucose load, were assessed at diagnosis of PP, in early puberty and after menarche; serum gonadotropins were measured in early puberty and after menarche; ovarian function was examined postmenarche. RESULTS: Comparisons of endocrine-metabolic results between normal- and low-birthweight PP girls showed no detectable differences before puberty. The hypertriglyceridaemia and elevated LDL-cholesterol levels characterizing low-birthweight PP girls became detectable by early puberty; reduced insulin sensitivity was not evident until postmenarche, when the tendency to ovarian dysfunction also became obvious. Body mass indices of normal- and low-birthweight subgroups were identical in early puberty and postmenarche. CONCLUSIONS: These longitudinal data show that, in PP girls, the endocrine-metabolic risk conferred by prenatal growth restraint is not readily detectable until puberty or postmenarche, and is not attributable to a higher body mass index. PMID- 11894978 TI - Lack of influence of the androgen receptor gene CAG-repeat polymorphism on sex steroid status and bone metabolism in elderly men. AB - OBJECTIVE: Population means for serum testosterone (T) levels in healthy men decrease with ageing but there is considerable interindividual variability of serum T in elderly men. Ultimate androgen action is mediated through the androgen receptor. Subtle differences in androgen sensitivity might contribute to serum T variability through the T negative feedback regulation. The androgen receptor gene (AR) contains in exon 1 a polymorphic trinucleotide CAG-repeat, whose length modulates androgen receptor action. The aims of the study were to assess the potential contribution of the AR CAG-repeat polymorphism in the interindividual variability of serum T and in the determination of bone metabolism in ambulatory elderly men. DESIGN AND PATIENTS: We used cross-sectional baseline data of a longitudinal study investigating the process of ageing, in particular the changes in hormonal status and bone metabolism, in a cohort of 273 community-dwelling healthy men, between age 71 and 86 years. MEASUREMENTS: AR CAG-repeat length was determined by automated DNA sequencing of exon 1 of the AR gene. Serum T, sex hormone binding globulin, LH and oestradiol were measured by specific immunoassays. Bone mineral density (BMD) was determined by dual energy X-ray absorptiometry. Bone turnover was assessed by measurement of serum bone-specific alkaline phosphatase, serum osteocalcin, serum C-terminal type I procollagen peptide, serum and urinary C-terminal telopeptides of type I collagen and urinary deoxypyridinoline levels, with use of immunoassays. RESULTS: No significant association was found between the AR CAG-repeat length and either total or free T, LH or the androgen sensitivity index (LHxT). BMD measurements at the hip and the forearm were not associated with AR CAG-repeat length and there was no association of this AR polymorphism with any of the biochemical markers of bone turnover. Results were not different after adjustments for age and body mass index. CONCLUSIONS: The findings of the present study do not support the view that in community-dwelling, healthy elderly men the androgen receptor gene CAG repeat polymorphism has a substantial impact on interindividual variability of serum testosterone levels or on the determination of bone turnover and bone mineral density. PMID- 11894980 TI - A placebo-controlled trial of long-term oral combined continuous hormone replacement therapy in postmenopausal women: effects on arterial compliance and endothelial function. AB - OBJECTIVE: To study the effects of long-term combined continuous oral hormone replacement therapy (HRT) on vascular function in healthy postmenopausal women. BACKGROUND: The cardiovascular effects of HRT are controversial. Improvement in vascular function is a proposed mechanism of oestrogen action but there are no long-term controlled human trials in this area. In this study, we examined the effects of HRT on lipid profiles and vascular function, encompassing both biomechanical arterial properties [systemic arterial compliance (SAC) and pulse wave velocity (PWV)] and endothelial function [flow-mediated vasodilation (FMD)]. METHODS: In this 2-year, double-blind, placebo-controlled study, 59 healthy postmenopausal women were randomized to oral combined continuous oestrogen and progesterone [Kliogest, oestradiol (2 mg), norethisterone (1 mg)] or placebo, with end-points measured at baseline, 6 weeks and after 6,12 and 24 months of treatment. RESULTS: Oral combined HRT reduced lipoprotein a [Lp(a)], although other lipid benefits were not observed. There were no significant changes in SAC, PWV or FMD with oral combined HRT, compared to placebo. CONCLUSION: In this long term, randomized placebo-controlled trial, oral continuous HRT with combined oestradiol and norethisterone in healthy postmenopausal women did not improve a spectrum of indices of arterial function compared to placebo. These results suggest that HRT might not be of cardiovascular benefit in postmenopausal women. PMID- 11894981 TI - Seasonal variation in glucocorticoid receptor binding characteristics in human mononuclear leucocytes. AB - OBJECTIVE: Glucocorticoid sensitivity varies between individuals and between tissues in the same individual. Although some of this variation is explained by the activity of the 11 beta-hydroxysteroid dehydrogenase enzymes, the possibility that glucocorticoid receptor sensitivity is modulated remains unexplored. This study examined glucocorticoid receptor binding in leucocytes and assessed the effects of seasonal hormonal variation on receptor binding. PATIENTS AND MEASUREMENTS: Two populations were studied. In the first, 318 healthy subjects were studied over 2 years with a single measurement of receptor binding made on each. In the second study nine healthy male subjects each had receptor binding measurements made at 3-week intervals over 1 year. RESULTS: In both populations there was significant seasonal variation in receptor binding. In the first population Kd for dexamethasone was highest in November and lowest in July (8.37 +/- 0.5 nmol/l vs. 1.58 +/- 0.7, mean +/- SEM P < 0.00001) and the number of receptor sites per leucocyte was highest in January and lowest in June (10 440 +/ 580 vs. 4969 +/- 302, P < 0.00001). In multivariate analysis, climate was the main determinant for both Kd and the number of receptor sites per cell: increases in day length and environmental temperature reduced Kd and the number of receptor sites per cell. Co-incubation with physiological concentrations of melatonin raised Kd without affecting receptor number. Co-incubation with forskolin lowered Kd suggesting that melatonin might act through the ML1 receptor class by inhibiting adenylyl cyclase. No correlations were found with 0900 h plasma cortisol. CONCLUSIONS: The results suggest that the glucocorticoid receptor might be modulated by season. Melatonin might mediate part of these effects. The lack of correlation with cortisol suggests that it is not an important determinant of receptor binding and that leucocyte receptors are regulated differently from central receptors. PMID- 11894982 TI - Malignant thymic carcinoid is not prevented by transcervical thymectomy in multiple endocrine neoplasia type 1. AB - Multiple endocrine neoplasia type 1 (MEN 1) is an autosomal dominant tumour syndrome. It is characterized by primary hyperparathyroidism, pituitary neoplasia and foregut lineage neuroendocrine neoplasia. Malignant thymic carcinoid tumours are an uncommon but important manifestation of MEN 1. Transcervical thymectomy is often advocated as prophylaxis against thymic carcinoids, although there is a paucity of evidence to support the efficacy of this procedure. This is the first report of a malignant thymic carcinoid occurring in an MEN 1 patient following prior parathyroidectomy and transcervical thymectomy. It is concluded that transcervical thymectomy does not reliably provide prophylaxis against thymic carcinoid. PMID- 11894983 TI - Comments on 'Sustained reduction in circulating cholesterol in adult hypopituitary patients given low dose titrated growth hormone replacement therapy: a two-year study' by Florakis et aL (2000). PMID- 11894984 TI - Choice of hormone replacement therapy in young women with ovarian failure. PMID- 11894985 TI - Photosynthesis-irradiance response at physiological level: a mechanistic model. AB - A mechanistic model is developed to present the photosynthetic response of phytoplankton to irradiance at the physiological level. The model is operated on photosynthetic units (PSU), and each PSU is assumed to have two states: reactive and activated. Light absorption that drives a reactive PSU into the activated state results from the effective absorption of the PSU. Transitions between the two states are asymmetrical in rate. A PSU in the reactive state becomes activated much faster than it recovers from the activated state to the reactive one. The turnover time for an activated PSU to transit into the reactive one is defined by the turnover time of the electron transport chain. The present model yields a photosynthesis-irradiance curve (PE-curve) in a hyperbola, which is described by three physiological parameters: effective cross-section (sigmaPSII), turnover time of electron transport chain (tau) and number of PSUs (N). The PE curve has an initial slope of sigmaPSII x N, a half-saturated irradiance of 1/(tau sigmaPSII), and a maximal photosynthetic rate of N/tau at the saturated irradiance. The PE-curve from the present model is comparable to the empirical function based on the target theory described by the Poisson distribution. PMID- 11894986 TI - The influence of selection on the evolutionary distance estimated from the base changes observed between homologous nucleotide sequences. AB - In most studies of molecular evolution, the nucleotide base at a site is assumed to change with the apparent rate under functional constraint, and the comparison of base changes between homologous genes is thought to yield the evolutionary distance corresponding to the site-average change rate multiplied by the divergence time. However, this view is not sufficiently successful in estimating the divergence time of species, but mostly results in the construction of tree topology without a time-scale. In the present paper, this problem is investigated theoretically by considering that observed base changes are the results of comparing the survivals through selection of mutated bases. In the case of weak selection, the time course of base changes due to mutation and selection can be obtained analytically, leading to a theoretical equation showing how the selection has influence on the evolutionary distance estimated from the enumeration of base changes. This result provides a new method for estimating the divergence time more accurately from the observed base changes by evaluating both the strength of selection and the mutation rate. The validity of this method is verified by analysing the base changes observed at the third codon positions of amino acid residues with four-fold codon degeneracy in the protein genes of mammalian mitochondria; i.e. the ratios of estimated divergence times are fairly well consistent with a series of fossil records of mammals. Throughout this analysis, it is also suggested that the mutation rates in mitochondrial genomes are almost the same in different lineages of mammals and that the lineage specific base-change rates indicated previously are due to the selection probably arising from the preference of transfer RNAs to codons. PMID- 11894987 TI - Selective imitation for a private sign system. AB - A distinctive feature of all human languages is the diverse and arbitrary nature of the sign (signifier). This can be interpreted as stating that the mapping between signals and referents is established by convention rather than by functional constraints. This property of the sign provides for a great deal of linguistic flexibility and is a key component of symbolic communication. Game theoretic models to describe signal imitation are investigated with a view to understanding how non-arbitrary (indexical) animal-style signals might 'evolve' culturally into diverse, arbitrary signs. I explore the evolutionary hypothesis that private, arbitrary signs emerge as a result of selective imitation within a socially structured population. Once arbitrary signs have emerged, they contribute towards greater assortative interactions among individuals using a shared sign system. In natural populations, the models for imitation will very often be close kin. Hence, kinship provides one mechanism for the creation of true symbols. An imitation-structured population can support many more sign systems than an equivalent non-structured population and is one in which symbols become the dominant force in assortative interactions. PMID- 11894988 TI - Analysis of a circular code model. AB - A circular code has been identified in the protein (coding) genes of both eukaryotes and prokaryotes by using a statistical method called trinucleotide frequency (TF) method [Arques & Michel (1996). J. theor. Biol. 182, 45-58]. Recently, a probabilistic model based on the nucleotide frequencies with a hypothesis of absence of correlation between successive bases on a DNA strand, has been proposed by Koch & Lehmann [(1997). J. theor. Biol. 189, 171-174] for constructing some particular circular codes. Their interesting method which we call here nucleotide frequency (NF) method, reveals several limits for constructing the circular code observed with protein genes. PMID- 11894989 TI - A cellular oscillator model for periodic pattern formation. AB - In this paper, we present a model for pattern formation in developing organisms that is based on cellular oscillators (CO). An oscillatory process within cells serves as a developmental clock whose period is tightly regulated by cell autonomous or non-autonomous mechanisms. A spatial pattern is generated as a result of an initial temporal ordering of the cell oscillators freezing into spatial order as the clocks slow down and stop at different times or phases in their cycles. We apply a CO model to vertebrate somitogenesis and show that we can reproduce the dynamics of periodic gene expression patterns observed in the pre-somitic mesoderm. We also show how varying somite lengths can be generated with the CO model. We then discuss the model in view of experimental evidence and its relevance to other instances of biological pattern formation, showing its versatility as a pattern generator. PMID- 11894990 TI - Can diving-induced tissue nitrogen supersaturation increase the chance of acoustically driven bubble growth in marine mammals? AB - The potential for acoustically mediated causes of stranding in cetaceans (whales and dolphins) is of increasing concern given recent stranding events associated with anthropogenic acoustic activity. We examine a potentially debilitating non auditory mechanism called rectified diffusion. Rectified diffusion causes gas bubble growth, which in an insonified animal may produce emboli, tissue separation and high, localized pressure in nervous tissue. Using the results of a dolphin dive study and a model of rectified diffusion for low-frequency exposure, we demonstrate that the diving behavior of cetaceans prior to an intense acoustic exposure may increase the chance of rectified diffusion. Specifically, deep diving and slow ascent/descent speed contributes to increased gas-tissue saturation, a condition that amplifies the likelihood of rectified diffusion. The depth of lung collapse limits nitrogen uptake per dive and the surface interval duration influences the amount of nitrogen washout from tissues between dives. Model results suggest that low-frequency rectified diffusion models need to be advanced, that the diving behavior of marine mammals of concern needs to be investigated to identify at-risk animals, and that more intensive studies of gas dynamics within diving marine mammals should be undertaken. PMID- 11894991 TI - A mathematical model of electron transfer within the mitochondrial respiratory cytochromes. AB - A simple mathematical model of electron flow along the mitochondrial respiratory cytochrome assembly and the transfer of electrons to molecular oxygen is presented. First, an expression for the current-voltage relationship for a biological oxygen electrode is derived, and from this the relationship between oxygen consumption rate and oxygen partial pressure is determined. An independent relationship between mitochondrial oxygen partial pressure and oxygen supply rate is then derived. By eliminating oxygen partial pressure from these two expressions, we may obtain a relationship between oxygen supply rate and oxygen consumption rate. This model is then used to investigate the effects of tissue dysoxia, uncoupling of oxidative phosphorylation, increased cellular diffusional resistance and inhomogeneities in oxygen supply on oxygen consumption. It is concluded that each of the above contribute in varying degrees to the phenomenon of "pathological oxygen supply dependency". PMID- 11894992 TI - Multiple asymmetry and concord resolutions of a conflict. AB - A model for a population-game with multiple asymmetry is studied, in which the participants are assumed to be different from one another both in size and in status as owners or non-owners of a territory. Only owners can reproduce, hence natural selection is assumed to operate in favor of the increase of ownership time. Conditions for the evolutionary stability of the Bourgeois Principle of owner-priority, despite difference in body size, are characterized. It is shown that ownership-priority tends to be at least partially replaced by strength priority as the availability of habitats, the expected longevity of potential intruders and the harm inflicted on the loser of an aggressive confrontation decrease, and as the expected longevity of the owner increases. It is further established that the combined effect of all these parameters can be characterized by a single parameter, referred to as the concord coefficient of the population. Finally, when this parameter reaches a certain critical level, only strength priority can prevail. If the concord coefficient decreases below this critical level, no priority-rule can remain stable in the population, in which case aggressive confrontations cannot be avoided, at least in certain situations. In this case, it is shown that aggression emerges first among low-rank individuals. PMID- 11894993 TI - Modelling mammary metabolism in the dairy cow to predict milk constituent yield, with emphasis on amino acid metabolism and milk protein production: model construction. AB - Previous efforts to simulate mammary metabolism have focused on energy, mostly considering amino acids (AA) in aggregate. The main objective of this work was to build a model of mammary metabolism, based on data from arterio-venous difference studies, which considered AA in sufficient detail to predict yields of milk solids. The model contains 19 state variables and considers the removal of 37 metabolites from blood, including 22 AA. It is driven by blood flow and arterial concentrations, and outputs include milk protein, milk lactose, and three classes of milk fat (by chain length). The model was parameterized using a balance version of it and the mean observations from four arterio-venous difference experiments, with a limited number of assumptions, and evaluated against these experiments. In assembling the balance model, milk protein output was not predicted satisfactorily, as some essential AA were not present in quantities great enough to support the rates of milk protein synthesis observed experimentally. Tryptophan showed the greatest deficit, followed by tyrosine plus phenylalanine, methionine, and histidine. In addition, significant quantities of pyruvate were needed to synthesize serine, glycine, and alanine. The supply of alpha-ketoglutarate plus glutamate to synthesize proline and glutamine was provided in part by catabolism of arginine; the remainder was derived from catabolism of other AA and energetic substrates. PMID- 11894995 TI - A theoretical investigation into the direct and indirect effects of state on the risk of predation. AB - As well as there being a direct physical effect of the state (for example fat reserves, or size) of an animal on the risk of being caught by a predator, state also has an effect on predation risk indirectly through changes in behaviour. We present a mathematical model which looks at these two components of the effect of state on predation risk. We focus on two different models, (i) where the animal must achieve a fixed state and its fitness depends on the time at which this state is reached and (ii) where the animal must survive until a fixed time and its fitness depends on its final state. We investigate conditions under which the indirect effect of increased state is to increase or decrease predation risk, and give some numerical illustrations. Under certain conditions in the fixed-state model, the indirect effect of state is to increase predation risk, whereas under certain conditions in the fixed-time model the indirect effect of state is to decrease predation risk. We discuss the implications of our results for empirical investigations into the effect of state on predation risk. PMID- 11894994 TI - The topology of the possible: formal spaces underlying patterns of evolutionary change. AB - The current implementation of the Neo-Darwinian model of evolution typically assumes that the set of possible phenotypes is organized into a highly symmetric and regular space equipped with a notion of distance, for example, a Euclidean vector space. Recent computational work on a biophysical genotype-phenotype model based on the folding of RNA sequences into secondary structures suggests a rather different picture. If phenotypes are organized according to genetic accessibility, the resulting space lacks a metric and is formalized by an unfamiliar structure, known as a pre-topology. Patterns of phenotypic evolution such as punctuation, irreversibility, modularity--result naturally from the properties of this space. The classical framework, however, addresses these patterns by exclusively invoking natural selection on suitably imposed fitness landscapes. We propose to extend the explanatory level for phenotypic evolution from fitness considerations alone to include the topological structure of phenotype space as induced by the genotype-phenotype map. We introduce the mathematical concepts and tools necessary to formalize the notion of accessibility pre-topology relative to which we can speak of continuity in the genotype-phenotype map and in evolutionary trajectories. We connect the factorization of a pre-topology into a product space with the notion of phenotypic character and derive a condition for factorization. Based on anecdotal evidence from the RNA model, we conjecture that this condition is not globally fulfilled, but rather confined to regions where the genotype-phenotype map is continuous. Equivalently, local regions of genotype space on which the map is discontinuous are associated with the loss of character autonomy. This is consistent with the importance of these regions for phenotypic innovation. The intention of the present paper is to offer a perspective, a framework to implement this perspective, and a few results illustrating how this framework can be put to work. The RNA case is used as an example throughout the text. PMID- 11894996 TI - Cooperative boundary populations: the evolution of cooperation on mortality risk gradients. AB - Cooperative or altruistic behavior is known to be vulnerable to destructive exploitation in the absence of spatial segregation and perceptual discrimination on the part of cooperators. In this study, a non-standard, agent-based, spatially explicit model of the evolution of cooperation shows that spatial gradients of increasing individual mortality risk can allow cooperative subpopulations to persist among players randomly matched for one-shot Prisoner's Dilemma. Further, the dynamically stable cooperator population formed on the gradient at the boundary of the survivable non-cooperative range provides ideal conditions for the evolution of discriminating strategies such as tit-for-tat. It is suggested that such gradients may commonly exist at the boundaries of the ranges of existing populations, providing a new basic mechanism for the evolution of cooperation. PMID- 11894997 TI - Genetically modified immunocompetent cells in HIV infection. AB - Even in the era of highly active antiretroviral therapy (HAART), gene therapy (GT) can remain a promising approach for suppressing HIV infection, especially if complemented with other forms of pharmacological and immunological intervention. A large number of vectors and targets have been studied. Here we discuss the potential of genetically treated, antigen-specific immunocompetent cells for adoptive autologous immunotherapy of HIV infection. Cellular therapies with gene modified CD8 and CD4 lymphocytes are aimed at reconstituting the antigen-specific repertoires that may be deranged as a consequence of HIV infection. Even if complete eradication of HIV from the reservoirs cannot be achieved, reconstitution of cellular immunity specific for opportunistic pathogens and for HIV itself is a desirable option to control progression of HIV infection and AIDS pathogenesis better. PMID- 11894998 TI - A phage display selected fab fragment with MHC class I-restricted specificity for MAGE-A1 allows for retargeting of primary human T lymphocytes. AB - The clinical benefit of adoptive transfer of MHC-restricted cytotoxic T lymphocytes(CTL) for the treatment of cancer is hampered by the low success rate to generate antitumor CTLs. To bypass the need for tumor-specific CTL, we developed a strategy that allows for grafting of human T lymphocytes with MHC restricted antigen specificity using in vitro selected human Fab fragments fused to the Fc(epsilon)RI-gamma signaling molecule. Retroviral introduction of a Fab based chimeric receptor specific for MAGE-A1/HLA-A1 into primary human T lymphocytes resulted in binding of relevant peptide/MHC complexes. Transduced T lymphocytes responded to native MAGE-A1/HLA-A1POS target cells by specific cytokine production and cytolysis. Therefore, peptide/MHC-specific Fab fragments represent new alternatives to TCR to confer human T lymphocytes with tumor specificity, which provides a promising rationale for developing immunogene therapies. PMID- 11894999 TI - Retrovirus-mediated transfer of prothymosin gene inhibits tumor growth and prolongs survival in murine bladder cancer. AB - To explore the potential use of prothymosin alpha(ProT), a putative thymic hormone, in gene therapy for bladder cancer, we generated a replication-defective recombinant retroviral vector encoding ProT and tested its antitumor effect on the MBT-2 murine bladder cancer. C3H/HeN mice injected with MBT-2 cells in conjunction with retroviruses encoding ProT exhibited smaller tumor mass, lower tumor incidence and higher survival rate, as well as higher antitumor cytotoxic activities compared with those injected with control viruses. However, such effects were not observed in severe combined immunodeficiency mice, suggesting that ProT exerts antitumor effects through its immunomodulatory activities. Cell growth in monolayer culture and colony formation in soft agar were enhanced in ProT gene-modified MBT-2 clones, and such growth-promoting activities of ProT could be reversed if its nuclear localization signal (NLS) was deleted. To circumvent the proliferation-promoting effect of ProT on tumor cells, a retroviral vector encoding ProT lacking NLS was constructed. Our results showed that retroviruses encoding NLS-deleted ProT was more efficacious than those encoding wild-type ProT in prolonging survival of tumor-bearing mice. This is the first report indicating that ProT, in particular NLS-deleted ProT, delivered by retroviral vectors may be further explored for the treatment of bladder cancer. PMID- 11895000 TI - Intra-arterial administration of a replication-selective adenovirus (dl1520) in patients with colorectal carcinoma metastatic to the liver: a phase I trial. AB - Both replication-incompetent and replication-selective adenoviruses are being developed for the treatment of cancer and other diseases. Concerns have been raised about the safety of intra-vascular adenovirus administration following a patient death on a clinical trial with a replication-defective adenovirus. In addition, the feasibility of vascular delivery to distant tumors has been questioned. dl1520 (ONYX-015) is a replication-selective adenovirus that has previously shown safety and antitumoral activity following intratumoral injection. This is the first report of intra-vascular administration with a genetically engineered, replication-selective virus. A phase I dose-escalation trial was performed in patients with liver-predominant gastrointestinal carcinoma (n = 11 total; primarily colorectal). dl1520 was infused into the hepatic artery at doses of 2 x 10(8)-2 x 10(1)2 particles for two cycles (days 1 and 8). Subsequent cycles of dl1520 were administered in combination with intravenous 5 fluorouracil (5-FU) and leucovorin. No dose-limiting toxicity, maximally tolerated dose or treatment-emergent clinical hepatotoxicity were identified following dl1520 infusion. Mild to moderate fever, rigors and fatigue were the most common adverse events. Antibody titers increased significantly in all patients. Viral replication was detectable in patients receiving the highest two doses. An objective response was demonstrated in combination with chemotherapy in a patient who was refractory to both 5-FU and dl1520 as single agents. Therefore, hepatic artery infusion of the attenuated adenovirus dl1520 was well-tolerated at doses resulting in infection, replication and chemotherapy-associated antitumoral activity. PMID- 11895001 TI - Intravascular injections of a conditional replicative adenovirus (adl118) prevent metastatic disease in human breast carcinoma xenografts. AB - We describe a study showing that the adenovirus adl118, lacking both E1B proteins, very efficiently kills human malignant cells 'in vitro' and 'in vivo'. Since many breast cancer patients do not have metastasis at the time of diagnosis, but finally develop it, we planned to study whether intravascular injections of adl118 could prevent metastatic development. We studied the effects of this mutant adenovirus in an orthotopic model of human breast carcinoma xenografts with the breast MB435-lung 2 cell line, which is highly metastatic in the lungs. In this study, all primary tumors were excised when they reached 50 100 mm3 volume in the animals. After surgery, 10(10) p.f.u. of adl118 was intravenously injected into a random group of animals, either three times during the first week only, or once every week. At death, almost all the control animals showed numerous lung metastases of large size, which were present in only 15-40% of the treated animals, depending on the size of the primary tumor at the time of excision. Overall survival was 50-70 days in control mice, and over 120 days in mice injected with adl118. Concomitant treatment with adl118 and cisplatin did not enhance the antitumor effects of adl118. With these results, we conclude that intravenous injection of conditional replicative adenovirus, after excision of the primary tumor, induces a clear decrease in the metastatic disease, and could be a new strategy in preventing tumor metastasis of breast carcinomas. PMID- 11895002 TI - Inhibition of intimal hyperplasia after balloon injury in rat carotid artery model using cis-element 'decoy' of nuclear factor-kappaB binding site as a novel molecular strategy. AB - The transcription factor, NFkB, plays a pivotal role in the coordinated transactivation of cytokine and adhesion molecule genes involved in atherosclerosis and lesion formation after vascular injury. We hypothesized that synthetic double-stranded DNA with high affinity for NFkB may be introduced as a 'decoy' cis element to bind the transcription factor, and block gene activation, resulting in an effective therapeutic agent for treating intimal hyperplasia. In vivo transfection of NFkB decoy oligodeoxynucleotides (ODN) into balloon-injured rat carotid artery resulted in the inhibition of neointimal formation at 14 days after injury as compared with vessels transfected with scrambled ODN (P < 0.01). It is of importance to note that in the vessels transfected with NFkB decoy ODN, the expression of p53, a pro-apoptotic gene, was upregulated in neointimal area, followed by increased apoptosis at 14 days. In addition, gene expression of ICAM 1 and VCAM-1 was markedly decreased in blood vessels transfected with NFkB decoy ODN compared with scrambled ODN, whereas balloon injury induced ICAM and VCAM expression in the neointimal area. More importantly, the migration of macrophages and T-lymphocytes into the neointima and media was significantly inhibited by NFkB decoy ODN as compared with scrambled ODN. Here, we demonstrated that in vivo transfer of NFkB decoy ODN successfully inhibited neointimal formation after balloon injury, accompanied by (1) induction of apoptosis through p53 upregulation, and (2) inhibition of local inflammatory actions through the downregulation of adhesion molecules. These results suggest that decoy treatment against NFkB provides a new therapeutic strategy to inhibit neointimal hyperplasia after angioplasty. PMID- 11895003 TI - The nuclear pore complex is involved in nuclear transfer of plasmid DNA condensed with an oligolysine-RGD peptide containing nuclear localisation properties. AB - One of the major barriers to efficient gene transfer and expression of nonviral vectors for gene therapy is passage across the nuclear envelope. We have previously shown that an oligolysine-RGD peptide that condenses plasmid DNA and binds to cell surface integrins can mediate increased internalisation of plasmid DNA into cells and synergistic enhancement of gene expression when complexed to a cationic lipid. In this report, we show that this enhancement is due to increased nuclear transfer of the plasmid DNA. We have applied the digitonin-permeabilised cell system that has been well established for the study of the nuclear transport of proteins to examine the nuclear transfer of plasmid DNA. Nuclear transfer of plasmid DNA complexed to an oligolysine-RGD peptide and lipofectamine appears to be an energy-dependent process involving the nuclear pore complex, since it is inhibited at 4 degrees C and by treatment with wheat germ agglutinin or with an antibody to the nuclear pore complex which all block nuclear protein import. In accordance with active nuclear transport, we have shown that all these treatments inhibit expression of a luciferase reporter plasmid in permeabilised cells. Nuclear transfer of pDNA is enhanced in mitotic cells, but cell division is not a prerequisite for transfer. We propose that the oligolysine-RGD peptide acts as a nuclear localisation signal and that the cationic lipid is more important for cell entry and endosome destabilisation than nuclear transfer. PMID- 11895004 TI - Enhanced suicide gene effect by adenoviral transduction of a VP22-cytosine deaminase (CD) fusion gene. AB - The low transduction efficiency of viral and nonviral vectors is a major limitation in tumour gene therapy. The HSV-1 tegument protein VP22 has been shown to exhibit a novel intercellular transport property. VP22 wild-type as well as VP22 fusion proteins efficiently spread from the original expressing cell to numerous neighbouring cells, so that protein transport by VP22 chimaeric polypeptides into the surrounding cells offers a possible compensation for the inadequate gene transfer efficiencies. To improve the therapeutic efficacy of the E. coli cytosine deaminase (CD) suicide gene we made use of the VP22 transport property in CD transducing adenoviral (Ad) vectors. C- and N-terminal fusions of CD linked in-frame with VP22 were generated and cloned into recombinant adenoviral vectors. Following in vitro transduction immunofluorescence analysis of Ad-transduced producer cells coplated with naive cells confirmed that the characteristic foci pattern of central producer and adjoining neighbour cells displaying nuclear staining was retained. After transduction of rat hepatoma cells with adenoviral vectors and subsequent incubation with the prodrug 5-FC, we observed enhanced cell cytotoxicity when comparing the CD-VP22 fusion (Ad-CD VP22) with Ad-vectors expressing the CD gene only (Ad-CD). Thereby employment of Ad-vectors encoding VP22 fusion proteins opens up new possibilities to potentiate the efficiency of suicide gene therapy for the treatment of solid tumours. PMID- 11895005 TI - In vivo gene transfer to the mouse eye using an HIV-based lentiviral vector; efficient long-term transduction of corneal endothelium and retinal pigment epithelium. AB - We have evaluated the transduction profiles of an HIV-based lentiviral vector delivered regionally to ocular tissues in vivo. Following subretinal injection, a green fluorescent protein (GFP) reporter gene was efficiently and stably expressed in retinal pigment epithelial (RPE) cells. Limited transduction of adjacent photoreceptors occurred in newborn mice, but was inefficient in adult animals. Injection of the vector into the anterior chamber resulted in efficient and stable transduction of corneal endothelial cells. Efficient in vivo gene transfer into cells of the corneal endothelium and retinal pigment epithelium by lentiviral vectors may therefore offer a valuable approach to the treatment of disorders of the cornea and outer retina. PMID- 11895006 TI - Microtubule involvement in the intracellular dynamics for gene transfection mediated by cationic liposomes. AB - The effects of microtubule polymerization on liposome-mediated gene transfection were investigated by confocal laser scanning microscopy in target living cells. Both nocodazole and taxol apparently increased the efficiency of gene transfection. Lipofection with fluorescence-labeled cationic liposomes in a COS-7 cell expressing yellow fluorescent protein (YFP)-tagged tubulin revealed that the liposomes were transported along microtubules to lysosomes which are colocalized with the microtubule organizing center (MTOC). Nocodazole disrupted microtubules and produced a uniform distribution of YFP-tagged tubulin in the cytoplasm. Under these conditions, both liposomes and lysosomes were scattered throughout the cytoplasm and they did not colocalize. In the presence of taxol, microtubules were stabilized and several focal regions, like the MTOC, were formed. Lysosomes resided around the nucleus, while liposomes were trapped in microtubules. Under these conditions, neither liposomes nor DNA colocalized with lysosomes. These results demonstrated that the liposome-DNA complexes are transported to lysosomes by a microtubule-mediated pathway, and the effects of nocodazole and taxol on transfection efficiency can be explained by failure of the transport of the liposome-DNA complexes to lysosomes where DNAs are degraded. PMID- 11895007 TI - Quo vadis? PMID- 11895009 TI - Diagnostic implications of stimulus polarity effects on the auditory brainstem response. AB - The purpose of this study was to determine whether clicks presented in rarefaction or condensation modes produce more accurate diagnostic information. Subjects were 20 consecutive patients who were seen at the Mayo Clinic for unilateral acoustic neuromas. The nontumor ear served as a control to minimize intersubject variability in the latencies. A standard audiologic evaluation was followed by an auditory brainstem response (ABR) test for which the stimuli were rarefaction and condensation clicks. Responses were analyzed for the presence of waves I, III, and V; absolute latencies of waves I, III, and V; interpeak intervals I-III, III-V, and I-V; and interaural latency difference for wave V. The results indicated that measures from both polarities were similar in this set of patients and that neither click polarity provided diagnostic advantages over the other. Recommendations are to collect ABRs to both click polarities individually to obtain the full complement of waves on which to base the diagnostic impression. PMID- 11895008 TI - Reversal of hemispheric asymmetry on auditory tasks in children who are poor listeners. AB - We examined hemispheric activation patterns during auditory and visual processing in two groups of children: 13 boys in the age range from 9 to 12 years rated by their parents and teachers as poor listeners and 11 boys in the same age range rated as normal listeners. Three tasks were employed: auditory gap detection, detection of auditory movement, and a control task involving visuospatial discrimination. Electrical activity was recorded from 30 scalp electrodes as participants responded to target stimuli in an event-related potential paradigm. In the visual task, hemispheric activation was relatively symmetric around the midsagittal plane in both groups. In the two auditory tasks, however, hemispheric activation patterns differed significantly between groups. In the normal-listener group, activation was asymmetric to the right hemisphere. In the poor-listener group, however, activation tended toward asymmetry, favoring the left hemisphere. These results suggest that abnormalities in hemispheric lateralization of function may underlie the auditory processing problems of at least some children described as poor listeners. PMID- 11895010 TI - Use of the median method to enhance detection of the mismatch negativity in the responses of individual listeners. AB - The median method was evaluated as an alternative way of expressing the mismatch negativity (MMN). Traditionally, signal averaging has been used to extract these event-related potentials from unwanted background noise. However, mean values are biased by unrejected artifact that skews the relatively small distribution of values on which the MMN is based. Because the median is a more valid measure of central tendency in asymmetric distributions, it may describe MMN data more accurately. Better representation of the signal in the median waveform might enhance detection of the MMN in the responses of individual listeners. Mean and median waveforms were computed from previously recorded MMN data. Visually identified MMNs were validated using area and onset latency criteria. Detectability of the MMN was not improved using median waveforms. Despite this result, a theoretical argument for use of the median is presented. PMID- 11895011 TI - The auditory processing battery: survey of common practices. AB - A survey of auditory processing (AP) diagnostic practices was mailed to all licensed audiologists in the State of Maryland and sent as an electronic mail attachment to the American Speech-Language-Hearing Association and Educational Audiology Association Internet forums. Common AP protocols (25 from the Internet, 28 from audiologists in Maryland) included requiring basic audiologic testing, using questionnaires, and administering dichotic listening, monaural low redundancy speech, temporal processing, and electrophysiologic tests. Some audiologists also administer binaural interaction, attention, memory, and speech language/psychological/educational tests and incorporate a classroom observation. The various AP batteries presently administered appear to be based on the availability of AP tests with well-documented normative data. Resources for obtaining AP tests are listed. PMID- 11895012 TI - Effects of diet and soil on the toxicity of lead in mallards. AB - Captive game-farm mallards (Anas platyrhynchos) were dosed with five No. 4 lead (Pb) shot and placed on a diet of shelled corn or commercial duck food (6 ducks). Half of the ducks on each diet was dosed daily with 10 g of soil. The commercial duck food provided more protection from the effects of Pb toxicity than the 10 g of soil dosed daily. Ducks on corn only died a mean of 7 days after dosing, ducks receiving commercial duck food, with and without soil, survived to the end of the study (21 days after dosing), and ducks receiving a diet of corn and soil survived an average of 17 days. Although ducks receiving corn only dissolved less Pb, excreted less Pb in their feces, and retained less dissolved Pb than the ducks receiving the other three treatments, they retained more dissolved Pb per day than the other ducks. PMID- 11895013 TI - Effects of substrate salinity on early seedling survival and growth of Scirpus robustus Pursh and Spartina alterniflora Loisel. AB - Rooted aquatic plants are being used increasingly to test the toxicity of sediments. However, effects of naturally occurring substrate constituents on most potential test species are not well understood even though their effects could affect the test results. The objective of this study was to determine the effect of substrate salinity (NaCl) on early seedling survival and growth of the emergent macrophytes, Scirpus robustus Pursh and Spartina alterniflora Loisel. Results of four 21- and 28-day toxicity tests, conducted in an artificial sediment, indicated interspecific differences in NaCl sensitivity when based on changes in shoot, root and whole plant dry-weight biomass. Concentrations of 7.8 g NaCl/l and 19.2 g NaCl/l first reduced early seedling biomass of S. robustus and S. alterniflora (P<0.05), respectively, when compared to plants grown in sediment containing no measurable salinity. Seedling survival was not affected at average concentrations of 17.5 g NaCl/l or less for S. robustus and 22.3 g NaCl/l or less for S. alterniflora. The results indicate that substrate salinity is an important consideration in the selection of test species for laboratory phytotoxicity tests conducted with estuarine sediments, particularly if determination of chronic toxicity attributable to anthropogenic contamination is the primary objective. PMID- 11895014 TI - Detection of cytogenetic alterations and blood cell changes in natural populations of carp. AB - A field study was conducted to investigate the appearance of alterations in the peripheral blood cells of wild populations of fish. Two aspects were evaluated: the appearance of cytogenetic effects, measured as increases on micronuclei frequencies, and the appearance of haematological effects by checking changes in the relative proportion of the different blood cell types. For this purpose common carp (Cyprinus carpio) were caught from four areas along a Spanish river. Three areas were located under the influence of chemical industries and/or a nuclear power plant. The fourth was a clean reference area. Flow cytometry was used to quantify the appearance of micronuclei on the same day of sampling and also after two and twelve months. The alterations in the relative proportion of cell types were counted in blood smears stained with Giemsa. Increases in micronuclei frequencies were observed in fish living in supposedly polluted areas. Alterations of the relative proportions of blood cells were manifested as an increase in white blood cells and as a decrease in red blood cells vs. control area. Since accidental spills have not been reported over this period of time, the alterations observed could suggest that fish are suffering chronic effects due to low level contamination associated with the sampled areas. PMID- 11895015 TI - Effects of temperature and salinity on life history of the marine amphipod Gammarus locusta. Implications for ecotoxicological testing. AB - The life history of Gammarus locusta was analysed in the laboratory under the following temperature and salinity combinations: 20 degrees C-33/1000, 15 degrees C-20/1000 and 15 degrees C-33/1000 (reference condition). Life history analysis comprised survival, individual growth, reproductive traits and life table parameters. Compared to 15 degrees C, life history at 20 degrees C was characterised by at least a four-week reduction in the life-span, lower life expectancy, shorter generation time, faster individual growth, anticipation of age at maturity and higher population growth rate. These temperature effects constituted an acceleration and condensation of the life cycle, compared to the reference condition. Concerning salinity effects, with few exceptions, results show that overall this amphipod life history did not differ significantly between the salinity conditions tested. Regarding ecotoxicological testing implications, findings from this study indicate that the range of temperature and salinity conditions acceptable for testing was substantially expanded both for acute and chronic assays. A temperature of 20 degrees C or higher (for a salinity of 33/1000) is suggested for routine chronic sediment toxicity testing with G. locusta, in order to reduce the life cycle and consequently improve cost effectiveness and standardisation. Results also suggest that a multiple-response approach, including survival, growth and reproduction, should be applied in chronic toxicity tests. PMID- 11895016 TI - The Human Genome Project (HGP). PMID- 11895017 TI - The post-genomic era and ecotoxicology. PMID- 11895018 TI - Placing Aboriginal and Torres Strait Islander mortality in an international context. AB - OBJECTIVE: To assess whether the patterns of mortality observed among Indigenous Australians were seen in other countries or sub-populations. Previous reports have indicated that the life expectancy of Indigenous Australians compares unfavourably with that of Indigenous groups in other developed countries, and is similar to that in some developing countries. However, in contrast to many developing countries, low life expectancy of Indigenous Australians is the result of relatively high and early adult mortality, rather than high infant mortality. METHODS: Using routinely available administrative data on age-specific mortality and estimated life expectancy at birth, we compared data for Indigenous Australians (from the Northern Territory, Western Australia and South Australia combined) with corresponding data for 200 countries world-wide, as well as for several population groups of interest, including African Americans, Native Americans, Canadian Natives and New Zealand Maori. RESULTS: Patterns of mortality among Indigenous Australians were markedly different to those of most other populations with available data, with the exception of the Russian Republic. The age-specific mortality rates for Russian males in 1990-95 were almost identical to those of Indigenous males in 1995-97. The similarities among females were less pronounced, but stronger than for any other country. CONCLUSION AND IMPLICATIONS: The dramatic fall in Russian life expectancy has been studied extensively and several important social and contextual factors have been identified. These factors are also relevant for the Indigenous population, and this may help to explain the similar mortality patterns of the two groups. PMID- 11895019 TI - Persistent social class mortality differences in New Zealand men aged 15-64: an analysis of mortality during 1995-97. AB - OBJECTIVE: Social class mortality differences in New Zealand men aged 15-64 years have previously been examined for the periods 1975-77 and 1985-87 using the Elley Irving social class scale. The objective was to repeat these analyses for 1995-97 in order to examine time trends, and to assess current social class patterns of mortality. METHODS: Age-standardised mortality rates were calculated for each social class and a weighted estimate of the social class mortality gradient was obtained. RESULTS: Male mortality declined 21% between 1985-87 and 1995-97, but the social class mortality differences have not diminished and may have even increased. The Relative Index of Inequality has increased from 1.8 in 1975-77 to 2.1 in 1985-87 and 2.3 in 1995-97. Unlike previous analyses, the relative social class mortality gradient was just as strong in the older age groups as in the younger age groups, indicating that the possible increase in social class gradient has largely occurred in the older age groups. CONCLUSIONS: These findings indicate that the potential to address the excess preventable mortality caused by socio-economic factors has not been fully realised in New Zealand. IMPLICATIONS: Social class analyses identify groups in the community that have an excess mortality that is potentially preventable. There are still major social class differences in mortality in New Zealand, and these differences may even have increased. It is important that these patterns are taken into account in public health planning and that further research is conducted to identify the mechanisms by which these differences occur. PMID- 11895020 TI - Particulate air pollution and hospital admissions in Christchurch, New Zealand. AB - AIMS: Winter air pollution in Christchurch is dominated by particulate matter from solid fuel domestic heating. The aim of the study was to explore the relationship between particulate air pollution and admissions to hospital with cardio-respiratory illnesses. METHODS: Particulate air pollution statistics (PM10) were obtained from the Canterbury Regional Council monitoring station in the city. The New Zealand Health Information Service provided data on admissions to the Princess Margaret and Christchurch Hospitals for the period June 1988 through December 1998 for both adults and children with cardiac and respiratory disorders. The relationship between PM10 and admissions was explored using a time series analysis approach controlling for weather variables. Missing values were interpolated from carbon monoxide data for the same time period, as carbon monoxide and PM10 were highly correlated. RESULTS: There was a significant association between PM10 levels and cardio-respiratory admissions. For all age groups combined there was a 3.37% increase in respiratory admissions for each interquartile rise in PM10 (interquartile value 14.8 mcg/m3). There was also a 1.26% rise in cardiac admissions for each interquartile rise in PM10. There was no relationship between PM10 and admissions for appendicitis, the control condition selected. CONCLUSIONS: In keeping with overseas studies, there is evidence in Christchurch of a relationship between ambient particulate levels and admissions with cardiac and respiratory illnesses. The size of the effect is consistent with overseas data, with the greatest impact for respiratory disorders. IMPLICATIONS: These results indicate that measures to control ambient particulate levels have the potential to reduce hospital admissions for cardio respiratory illnesses. PMID- 11895021 TI - A program for research and publication. PMID- 11895022 TI - Social determinants of smoking among parents with infants. AB - OBJECTIVES: To estimate the smoking prevalence among parents of infants and examine these parents' socio-demographic characteristics. METHOD: The sample of all parents of infants (669 mother-father pairs, 90 single parents) was derived from the 1995 Australian Health Survey. Data were collected by face-to-face interview in the respondent's home. Socio-demographic measures include parent's age, family structure, age-left-school, highest post-school qualification, occupation, and family income. RESULTS: The overall rate of smoking among parents was 28.9% (mothers 24.7%, fathers 33.7%). The lowest rate was observed among mothers with a post-school tertiary qualification (7.6%) and the highest among fathers aged 18-24 (49.0%). In 15.4% of two-parent families both parents smoked, but this rate differed markedly by family income (9.9% vs. 29.7% for high and low income families respectively). Multiple logistic regression showed that parents who smoked were more likely to be young, minimally educated, employed in blue collar occupations, and resident in low-income families. CONCLUSIONS AND IMPLICATIONS: Infants in this sample who were exposed to parental smoking were likely to be at increased risk of experiencing higher mortality and morbidity for childhood conditions related to passive smoking; more likely to experience adverse health consequences in adulthood; and may themselves take up smoking in later life. The study results pose serious challenges to our tobacco control efforts and health interventions more generally. No single policy or strategy can adequately address the problem of parental smoking. We need macro/upstream approaches that deal with the degree of social and economic inequality in society, as well as more intermediate approaches that intervene at the level of communities, families and individuals. PMID- 11895023 TI - Evaluation of smoke-free dining in South Australia: support and compliance among the community and restaurateurs. AB - OBJECTIVES: To evaluate the impact of smoke-free dining legislation introduced in South Australia on 4 January 1999. Specifically, to determine changes in public support and support among the restaurant and cafe industry; to determine changes to self-reported experiences of dining out; and to assess compliance with the legislation. METHODS: A series of independent, representative, cross-sectional surveys were undertaken over time. Study 1: 3000+ South Australians, aged 15 and over, were interviewed about opinions of the legislation and about dining out, in an annual household survey in October-November in 1997, 1998 and 1999. Study 2: 500+ owners and managers of public dining venues were interviewed about their opinions about and experiences of the legislation in face-to-face surveys in May June 1999 and July 2000. RESULTS: Study 1: Public support for smoke-free dining increased from 73.4% in 1997 to 84.2% in 1999. In 1999, 60.2% reported the ban had made dining out more enjoyable and 35.1% indicated no difference. Study 2: 88.2-92.3% of restaurants were complying with the legislation at five months and 95.7-99.6% at 18 months. In 2000, 82% of restaurateurs reported that they had spent no money in order to implement the law. CONCLUSIONS AND IMPLICATIONS: Smoke free dining in South Australia is popular and has resulted in increased perceived enjoyment for patrons. The legislative change has apparently been adhered to by both the majority of restaurateurs and customers, and was inexpensive for restaurateurs to implement. Smoke-free dining legislation can be implemented with confidence. PMID- 11895024 TI - Retrospective hospital-based searches for cases of acute flaccid paralysis. AB - OBJECTIVE: Australia had to demonstrate adequate acute flaccid paralysis (AFP) surveillance by achieving a rate of one per 100,000 in children under the age of 15 to fulfil one of the requirements of the Regional Commission for the Certification of Poliomyelitis Eradication to be declared polio free. To increase the ascertainment rate of AFP cases, a hospital search was conducted to identify cases not reported to the active AFP surveillance. METHODS: A computerised search of hospital admissions in New South Wales (NSW) and Western Australia (WA) on ICD 9 codes of Guillain Barre Syndrome (GBS), unspecified encephalitis, poliomyelitis, vaccine-associated paralytic polio (VAPP) and flaccid paralysis was conducted for the period 1995-98. Medical records of cases that were not reported to the active surveillance were reviewed in three hospitals of NSW and two hospitals in WA. RESULTS: Twenty additional cases recorded as GBS and five as transverse myelitis (TM) were identified through the searches, which increased the average four-year AFP rate from 1.0 to 1.4 per 100,000 in children under the age of 15 years in these two states and the overall AFP rate in Australia increased from 0.78 to 1.14. There were no cases of polio or VAPP found. Nine cases of GBS and five of TM reported to the active AFP surveillance were not found in the hospital searches. CONCLUSION: A combination of active surveillance and hospital-based searches increased the investigated AFP rate, which fulfilled one of the certification requirements for Australia to be certified polio free. IMPLICATIONS: Until global certification is achieved, AFP surveillance needs to be improved to identify cases of importation of wild poliovirus. PMID- 11895025 TI - A personal reflection on election issues and lessons. PMID- 11895026 TI - Measuring physical activity in public open space--an electronic device versus direct observation. AB - OBJECTIVE: To evaluate the practicality and validity of using infra-red beam counters (IRBCs) to measure walking path use and overall park use. METHOD: Direct observation was carried out simultaneously with IRBC data collection in five parks on seven non-consecutive days during February-March in 1998 and on matched days in 1999. A second validation study was undertaken in one park in October 1999. RESULTS: The IRBC over-estimated the observed number of people using walking paths by 14% to 78%. When assessed by path volume, the difference between the IRBC and observer ranged from 10% under-estimation to 16% over-estimation. In a more rigorous evaluation of path volume the IRBC under-estimated the observed count by 20%. The extent to which the IRBC equated with the number of observed park users varied from 69% under-estimation to no difference. CONCLUSION: IRBCs are not appropriate for measuring the number of people engaging in physical activity in parks. IMPLICATIONS: IRBCs cannot replace direct observation for the collection of valid data on physical activity participation in parks. Further research is needed to determine settings in which electronic devices such as IRBCs may provide valid data on physical activity participation. PMID- 11895027 TI - Validity and utility of community health workers' estimation of kava use. AB - OBJECTIVE: Estimating illicit substance use in epidemiological studies is challenging, particularly across ethical, cultural and language barriers. While developing the methods for a case-control study of the effects of heavy kava consumption among Aboriginal people in remote Northern Territory (NT), we examined the validity and utility of alternative methods for estimating exposure. METHODS: We assessed the level of agreement between a consensus of Aboriginal health workers in two different communities using interviews conducted with community members and health workers and individuals' self-reported kava consumption. Exposure measures included history of kava use, current kava use and history of heavy use. Agreement between a health worker consensus classification and individuals' self-report was analysed and agreement among several health workers in a consensus classification without self-report was assessed. RESULTS: Health workers concurred about an individual's history of kava use (k=0.83), current use (k=0.43) and also level of use (k=0.33). There was very good agreement between health workers' consensus and self-reported history of use (k=0.77). Agreement among health workers about current kava use was poor (k=0.08), while there was fair agreement between health workers and self-reported heavy kava users (k=0.36). Data from review of clinic patient notes supported agreement between consensus classification and self-reported history and level of use (k=0.39). CONCLUSIONS: Self-reported kava use may be a poor estimate of current use especially when obtained from interviews away from a confidential clinic setting. Consensus classification by knowledgeable Aboriginal health workers provided comprehensive coverage, efficiently and with greater reliability and assisted to identify 'excessive' kava use. PMID- 11895028 TI - Increases in vaccination coverage for children in child care, 1997 to 2000: an evaluation of the impact of government incentives and initiatives. AB - OBJECTIVE: To compare vaccination coverage of children in child care before (1997) and after (2000) implementation of government immunisation strategies including parent/providers incentives and surveillance of vaccination uptake. METHODS: Cross-sectional parent surveys of vaccination coverage for children (<3 years old regularly attending child care) in 47 child care centres and 19 councils operating family day care in metropolitan Melboume. RESULTS: Data were collected for 1,578 (72%) children in 1997 and 1,793 (72%) in 2000. In 2000, 93% were completely immunised, a 9% (95% CI 6%-11%, p<0.001) increase from 1997. Less than 1% of children were unimmunised (0.8% in 1997, 0.5% in 2000). For those >2 years, 94% were completely immunised before their second birthday in 2000 compared with 80% in 1997. Immunisation levels were 10% (95% Cl 6-12) higher in 2000 than in 1997 for those receiving child care benefits compared with a 7% (95% Cl 3-10) increase for families not receiving benefits. In 1997, 8 (17%) child care centres and 4 (21%) councils reported > or = 90% children completely immunised increasing to 33 (70%) and 16 (84%) in 2000 respectively. Fewer families reported delaying immunisations because of minor illness in 2000 (27%) compared to 1997 (44%, p<.001). Updating immunisation data by child care coordinators increased from 51% in 1997 to 98% in 2000. CONCLUSION: A substantial increase in immunisation uptake has been achieved for this population of young children attending child care. This study provides evidence that the increase in vaccination rates is attributable to some extent to increased surveillance of immunisation rates and both parent and provider incentives to immunise. PMID- 11895029 TI - Research should not ignore the power of elite groups. AB - For a long time, public health research has focused on many and diverse types of vulnerable groups in society. Whether drug abusers, prison inmates, smokers, poverty-stricken, mentally ill, HIV positive, or any number of other ailments, the public health system and, in particular, researchers have addressed and reported on people and populations with so-called deficits. Research on these vulnerable groups has been designed, conducted, reported, discussed and analysed, but vulnerable groups continue and, in fact, appear to be increasing in number, size, and range. An alternative approach that begins with entirely different questions is possible and advisable. Public health research could, and in the author's point of view, should, examine the lifestyles and, specifically, decision-making by members of elite groups. Studying up the economic and political ladder calls for examination of how a few reap enormous benefits for themselves, leaving the rest of us to our misfortune. Such decisions as the elite groups make affects resource allocation, use of information, and results in the public health and other social, economic, military, and political sectors in society. PMID- 11895030 TI - Poor health care worker vaccination coverage and knowledge of vaccination recommendations in a tertiary Australia hospital. AB - OBJECTIVES: Guidelines for vaccination of health care workers (HCWs) have been available in Victoria since 1998. We estimated knowledge and attitudes towards vaccination among HCWs as well as self-reported vaccination status in a tertiary adult hospital in Melbourne, Australia. METHODS: A cross-sectional survey was conducted in July 2000 using a telephone questionnaire and proportionate random sampling. RESULTS: Only 18% of 269 HCWs were fully vaccinated. Most (76%) had not heard of or seen current guidelines for HCW vaccination despite a stated belief in the importance of full vaccination (94%) and a willingness to update their vaccination status if necessary (96%). Less than half kept vaccination records (39%). Hepatitis B vaccination (95%) was most commonly completed. However, only half of all HCWs had received influenza vaccination in the past 12 months and other vaccines often had suboptimal coverage. A common reason cited for avoiding vaccination was concern over vaccine side effects (17%). While the hospital staff clinic was an acceptable site for vaccination, improved access was seen as important. CONCLUSIONS: HCW vaccination coverage and knowledge of vaccination requirements were poor. Concerns about vaccine side effects were common. IMPLICATIONS: Adequately resourced HCW vaccination programs that include ongoing education for HCWs and improved access to vaccination are necessary to improve vaccination coverage and reduce the risk of vaccine-preventable diseases among staff and patients. PMID- 11895031 TI - The risk of Ross River and Barmah Forest virus disease in Queensland: implications for New Zealand. AB - OBJECTIVES: To describe the incidence of Ross River (RR) and Barmah Forest (BF) virus disease in Queensland and determine the risk of importation of RR virus from Queensland into New Zealand (NZ) via viraemic travellers. METHODS: Based on routine RR and BF virus notification data of seven major urban tropical and subtropical Queensland populations, incidence rates adjusted for age, sex, season and a baseline level of immunity were used to examine the annual and seasonal risk of disease in the specific populations and selected subgroups. The risk for NZ was determined by estimating the number of infections among major visitor groups travelling from Queensland to NZ, using seroconversion rates. RESULTS: In Queensland, annual rates of RR and BF virus disease ranged between 31.5-288.3 and 3.4-37.4/100,000 person years respectively and increased to between 48.4-423.5 and 3.8-40.4/100,000 person years at risk when adjusted for immunity. Our estimates indicate that more than 100 viraemic travellers may enter NZ from Queensland each year. Estimates were greatest among New Zealanders returning home. CONCLUSIONS AND IMPLICATIONS: Usefulness of notification data could be maximised by presenting more detailed information to the local governments responsible for the control and public health awareness of these pathogens. Given the high number of viraemic persons entering NZ, the abundance of possums and the emergence of Oc. camptorhynchus, transmission of RR virus within NZ is probable. Health authorities should prepare for a virgin soil epidemic of RR virus by initiating serological and clinical surveillance in key areas, enhance public and professional awareness and elevate national resources necessary to invoke emergency vector control and case management. PMID- 11895032 TI - Changes in smoke-free home status in an immigrant Lebanese community in Sydney, Australia. PMID- 11895033 TI - Severe vein damage caused by Temezepam injecting. PMID- 11895034 TI - Robin Hood Index under discussion. PMID- 11895035 TI - Health issues of asylum seekers and refugees. AB - This paper is written on behalf of the West Australian Branch of the Australasian Faculty of Public Health Medicine. As public health physicians, we feel it is important that public health professionals should contribute constructively to address the needs of a socially deprived, marginalised group with high rates of physical and psychiatric morbidity. Depending on the definition, there are between 18 and 48 million asylum seekers and refugees in the world. Most seek protection in neighbouring countries, largely in Africa and Asia, rather than coming to North America, Europe and Australasia. Contrary to popular belief, numbers of successful applications to Australia's humanitarian program have actually fallen. This article attempts to correct misperceptions and misapprehensions about the effect of asylum seekers on the public health. Public health professionals should lobby for changes to Govemment policy that at present leave asylum seekers vulnerable to a cycle of poverty, ill-health and limited access to health services. PMID- 11895036 TI - The WHO classification of tumors of the nervous system. AB - The new World Health Organization (WHO) classification of nervous system tumors, published in 2000, emerged from a 1999 international consensus conference of neuropathologists. New entities include chordoid glioma of the third ventricle, cerebellar liponeurocytoma, atypical teratoid/rhabdoid tumor, and perineurioma. Several histological variants were added, including tanycytic ependymoma, large cell medulloblastoma, and rhabdoid meningioma. The WHO grading scheme was updated and, for meningiomas, extensively revised. In recognition of the emerging role of molecular diagnostic approaches to tumor classification, genetic profiles have been emphasized, as in the distinct subtypes of glioblastoma and the already clinically useful 1p and 19q markers for oligodendroglioma and 22q/INI1 for atypical teratoid/rhabdoid tumors. In accord with the new WHO Blue Book series, the actual classification is accompanied by extensive descriptions and illustrations of clinicopathological characteristics of each tumor type, including molecular genetic features, predictive factors, and separate chapters on inherited tumor syndromes. The 2000 WHO classification of nervous system tumors aims at being used and implemented by the neuro-oncology and biomedical research communities worldwide. PMID- 11895037 TI - Cytonectin expression in Alzheimer disease. AB - Cytonectin is a novel 35,000 molecular weight protein that displays remarkable ion-independent adherence properties. This consigns it to a family of well-known adherence molecules essential for cell communication and the development of 3 dimensional tissue structures. Cytonectin is expressed in a variety of organs and tissues, being evolutionarily conserved from human to avian species. It is hypothesized to serve as a key structural component of the body, and as a "do not attack" signal molecule that prevents tissue destruction by cells of monocyte lineage. This paper describes the properties of cytonectin and its proposed role in normal and disease states. The protein is overexpressed in Alzheimer disease entorhinal cortex as compared to normal age-matched controls. It is also detected in tissues from patients with Down syndrome and leukemia. Its presence in all 3 of these related conditions may prove important to their etiopathogenesis. PMID- 11895038 TI - Expression of phosphatidylserine receptor and down-regulation of pro-inflammatory molecule production by its natural ligand in rat microglial cultures. AB - Exposure of phosphatidylserine (PS), an aminophospholipid normally sequestered in the inner leaflet of plasma membrane, is one of the crucial steps in the recognition and ingestion of apoptotic cells by macrophages. The recognition of PS on apoptotic cells by peripheral macrophages is mediated by a phosphatidylserine-specific receptor (PtdSerR), which has recently been cloned. In spite of the important role of apoptosis in the CNS, the process of apoptotic neuron recognition by microglia is poorly understood. Because recent studies suggest that engagement of PS with a not yet characterized microglial receptor is necessary for apoptotic neuron uptake, we investigated the expression of PtdSer-R and its functional role in neonatal rat brain microglial cultures. Semi quantitative RT-PCR analysis revealed that PtdSerR mRNA was detectable in unstimulated cultures and enhanced in LPS activated microglia. The presence of PS liposomes strongly reduced the release of pro-inflammatory molecules such as nitric oxide, interleukin-1beta, and tumor necrosis factor-alpha by LPS-activated microglia. At variance, the immunoregulatory cytokines interleukin-10 and transforming growth factor-beta1 were moderately decreased or unaffected. The activity of PS-liposomes was mimicked by the PS head group phospho-L-serine, but not by phosphatidylcholine-containing liposomes. Our data suggest that, as for peripheral macrophages, PS through its receptor can modulate microglial activation toward an anti-inflammatory phenotype. PMID- 11895039 TI - 14-3-3 proteins in Lewy bodies in Parkinson disease and diffuse Lewy body disease brains. AB - Several components of Lewy bodies have been identified, but the precise mechanism responsible for the formation of Lewy bodies remains undetermined. The 14-3-3 protein family is involved in numerous signal transduction pathways and interacts with alpha-synuclein, which is a major constituent of Lewy bodies. To elucidate the role of 14-3-3 proteins in neuro-degenerative disorders associated with Lewy bodies, we performed immunohistochemical studies on 14-3-3 in brains from 5 elderly control subjects and from 10 patients with Parkinson disease (PD) or diffuse Lewy body disease (DLBD). In the normal controls, 14-3-3-like immunoreactivity was mainly observed in the neuronal somata and processes in various cortical and subcortical regions. In the PD and DLBD cases, a similar immunostaining pattern was found and immunoreactivity was generally spared in the surviving neurons from the severely affected regions. In addition, both classical and cortical Lewy bodies were intensely immunolabeled and some dystrophic neurites were also immunoreactive for 14-3-3. Our results suggest that 14-3-3 proteins may be associated with Lewy body formation and may play an important role in the pathogenesis of PD and DLBD. PMID- 11895040 TI - Familial Danish dementia: a novel form of cerebral amyloidosis associated with deposition of both amyloid-Dan and amyloid-beta. AB - Familial Danish dementia (FDD) is pathologically characterized by widespread cerebral amyloid angiopathy (CAA), parenchymal protein deposits, and neurofibrillary degeneration. FDD is associated with a mutation of the BRI2 gene located on chromosome 13. In FDD there is a decamer duplication, which abolishes the normal stop codon, resulting in an extended precursor protein and the release of an amyloidogenic fragment, ADan. The aim of this study was to describe the major neuropathological changes in FDD and to assess the distribution of ADan lesions, neurofibrillary pathology, glial, and microglial response using conventional techniques, immunohistochemistry, confocal microscopy, and immunoelectron microscopy. We showed that ADan is widely distributed in the central nervous system (CNS) in the leptomeninges, blood vessels, and parenchyma. A predominance of parenchymal pre-amyloid (non-fibrillary) lesions was found. Abeta was also present in a proportion of both vascular and parenchymal lesions. There was severe neurofibrillary pathology, and tau immunoblotting revealed a triplet electrophoretic migration pattern comparable with PHF-tau. FDD is a novel form of CNS amyloidosis with extensive neurofibrillary degeneration occurring with parenchymal, predominantly pre-amyloid rather than amyloid, deposition. These findings support the notion that parenchymal amyloid fibril formation is not a prerequisite for the development of neurofibrillary tangles. The significance of concurrent ADan and Abeta deposition in FDD is under further investigation. PMID- 11895041 TI - The pathology of the spinal cord in progressive supranuclear palsy. AB - We describe the results of a study of the spinal cord of 5 patients with progressive supranuclear palsy (PSP). Examination of the 6th cervical, 7th thoracic, and 5th lumbar segments revealed variable degree of gliosis and density of neuropil threads (NTs), nerve cell loss, and tau-positive cytoplasmic staining of neurons, some of which was reminiscent of neurofibrillary tangles (NFT). Tau positive neurons were seen at each spinal level and in the 3 zones in which each level was subdivided. Cells with the appearance of NFT predominated in the intermediate zone. Morphometric study revealed 47%, 52%, and 32% decrease in cell numbers in the motor area (lamina IX) at the 3 spinal levels, respectively, and 39% in the intermedio-lateral column. This is the first report describing severe neuronal loss throughout the whole spinal cord in patients with PSP and its results are in keeping with a previous study of the nucleus of Onufrowicz. The reasons why cell loss fails to produce clinical symptoms are analyzed and the clinico-pathological correlations between anatomical changes and dystonia are considered. On the basis of existing data, we conclude that previous suggestions implicating spinal interneurons in the pathogenesis of neck dystonia should not be supported. PMID- 11895042 TI - Differential expression between pilocytic and anaplastic astrocytomas: identification of apolipoprotein D as a marker for low-grade, non-infiltrating primary CNS neoplasms. AB - Fibrillary astrocytoma, the most common primary central nervous system neoplasm, is infiltrating, rapidly proliferating, and almost invariably fatal. This contrasts with the biologically distinct pilocytic astrocytoma, which is circumscribed, often cystic, slowly proliferating, and associated with a favorable long-term outcome. Diagnostic markers for distinguishing pilocytic astrocytomas from infiltrating anaplastic astrocytomas are currently not available. To identify genes that might either serve as markers or explain these distinct biologic behaviors, cDNA microarray analysis was used to compare the expression of 7,073 genes (nearly one quarter of the human genome) between these 2 types of astrocytoma. Messenger RNAs pooled from 3 pilocytic astrocytomas and from 4 infiltrating anaplastic astrocytomas were compared. Apolipoprotein D (apoD), which expressed 8.5-fold higher in pilocytic astrocytomas, showed the greatest level of differential expression and emerged as a potential marker for pilocytic tumors. By immunohistochemistry, 10 of 13 pilocytic astrocytomas stained positively for apoD, while none of 21 infiltrating astrocytomas showed similar staining. ApoD immunostaining was also seen in 9 of 14 of gangliogliomas, 4 of 5 subependymal giant cell astrocytomas (SEGAs), and a single pleomorphic xanthoastrocytomas (PXAs). By in situ hybridization, pilocytic astrocytomas, in contrast with infiltrating astrocytomas, showed widespread increased apoD expression. SAGE analysis using the NCBI database showed a higher level of expression of apoD RNA in pilocytic astrocytoma than in any of the other 94 neoplastic and non-neoplastic tissues in the database. ApoD is associated with decreased proliferation in some cell lines, and is the protein found in highest concentration in cyst fluid from benign cystic disease of the breast. ApoD might play a role in either decreased proliferation or cyst formation in pilocytic astrocytomas, gangliogliomas, SEGAs, and PXAs. PMID- 11895043 TI - Two types of sporadic cerebral amyloid angiopathy. AB - Cerebral amyloid angiopathy (CAA) is a type of beta-amyloidosis that occurs in leptomeningeal and cortical vessels of the elderly. In a sample of 41 CAA cases including 16 Alzheimer disease (AD) cases and 28 controls, we show that 2 types of sporadic CAA exist: The first type is characterized by immunohistochemically detectable amyloid beta-protein (Abeta) in cortical capillaries, leptomeningeal and cortical arteries, arterioles, veins, and venules. It is referred to here as CAA-Type 1. The second type of CAA also exhibits immunohistochemically detectable Abeta deposits in leptomeningeal and cortical vessels, with the exception of cortical capillaries. This type is termed CAA-Type 2. In cases with CAA-Type 1, the frequency of the apolipoprotein E (ApoE) epsilon4 allele is more than 4 times greater than in CAA-Type 2 cases and in controls. CAA-Type 2 cases have a higher epsilon2 allele frequency than CAA-Type 1 cases and controls. The ratio of CAA Type 2 to CAA-Type 1 cases does not shift significantly with respect to the severity of AD-related beta-amyloidosis, with respect to degrees of CAA-severity, or with increasing age. Therefore, CAA-Type 1 is unlikely to be the late stage of CAA-Type 2; rather, they represent 2 different entities. Since both the ApoE epsilon2 and the epsilon4 allele are known to be risk factors for CAA, we can assign the risk factor ApoE epsilon4 to a distinct morphological type of CAA. The ApoE epsilon4 allele constitutes a risk factor for CAA-Type 1 and, as such, for neuropil-associated dyshoric vascular Abeta deposition in capillaries, whereas the e2 allele does not. CAA-Type 2 is not associated with the epsilon4 allele as a risk factor but shows a higher epsilon2 allele frequency than CAA-Type 1 cases and controls in our sample. PMID- 11895044 TI - Commentary on "Synaptic Pathology in Prefrontal Cortex Is Present only with Severe Dementia in Alzheimer Disease" (JNEN 2001; 60;929-36). PMID- 11895045 TI - Hepatitis B vaccination safety. AB - BACKGROUND: Recent studies have suggested that adult hepatitis B vaccination may be associated with adverse reactions. OBJECTIVE: To further examine the relative risk, percentage association, and statistical significance of arthritic, immunologic, and gastrointestinal adverse reactions reported after adult hepatitis B vaccination compared with control vaccines. DESIGN: The Vaccine Adverse Events Reporting System (VAERS) database was analyzed for the incidence of adverse reactions after adult hepatitis B immunization compared with the incidence of adverse reactions reported to VAERS about vaccine control groups. SETTING: The medical and scientific communities have generally accepted that hepatitis B vaccine, a highly purified, genetically engineered single-antigen vaccine, is a safe vaccine. METHODS: The VAERS database was analyzed from 1997 to 2000 for adverse reactions associated with adult hepatitis B vaccination and from 1991 to 2000 for adverse reactions reported about vaccine control groups. RESULTS: The results showed a statistically significant increase in the incidence of adverse reactions reported after adult hepatitis B vaccination when compared with the incidence of adverse reactions reported to VAERS about control vaccines. CONCLUSIONS: Patients and physicians need to be fully informed of the potential adverse reactions associated with hepatitis B vaccination so that together they can make an informed consent decision about the risk versus the benefit. Patients who may have had an associated adverse reaction to hepatitis B vaccine should be made aware that they may be eligible for compensation from the no-fault Vaccine Compensation Act, administered by the US Court of Claims. PMID- 11895046 TI - Effects of Panax ginseng on quality of life. AB - OBJECTIVE: To assess the time-dependent effects of Panax ginseng on health related quality of life (HRQOL) by use of a general health status questionnaire. METHODS: Subjects were randomized in a double-blind manner to P. ginseng 200 mg/d (n = 15) or placebo (n = 15) for 8 weeks. The Short Form-36 Health Survey version 2 (SF-36v2), a validated general health status questionnaire, was used to assess HRQOL at baseline and at 4 and 8 weeks. HRQOL between the groups was compared by use of repeated-measures analysis of covariance. A p value <0.05 was considered statistically significant. RESULTS: There were no significant differences in baseline demographics and SF-36v2 scores between the groups. After 4 weeks of therapy, higher scores in social functioning (P. ginseng 54.9+/-4.6 vs. placebo 49.2+/-6.5; p = 0.014), mental health (P. ginseng 52.2+/-7.7 vs. placebo 47.2+/ 7.3; p = 0.075), and the mental component summary (P. ginseng 51.3+/-7.4 vs. placebo 44.3+/-8.3; p = 0.019) scales were observed in patients randomized to P. ginseng; these differences did not persist to the 8-week time point. The incidence of adverse effects was 33% in the P. ginseng group compared with 17% in the placebo group (p = 0.40). Subjects given P. ginseng (58%) were more likely to state that they received active therapy than subjects given placebo (17%; p < 0.05). CONCLUSIONS: P. ginseng improves aspects of mental health and social functioning after 4 weeks of therapy, although these differences attenuate with continued use. PMID- 11895047 TI - Accuracy of the avosure PT pro system compared with a hospital laboratory standard. AB - OBJECTIVE: To compare international normalized ratio (INR) values obtained using the AvoSure PT Pro point-of-care (POC) system with those obtained using a standard laboratory method. METHODS: Forty-one INR values obtained from the POC system were compared with those obtained from a standard laboratory method. The POC method was evaluated for both laboratory and clinical agreement. To evaluate laboratory agreement, various analyses were used, including mean-squared prediction error (MSE) and mean prediction error (ME), Bland-Altman analysis, correlation, and paired t-test comparing group INR means. For clinical accuracy, discrepant pairs were identified and evaluated to determine whether dosage adjustments would have been needed based on values obtained. RESULTS: The POC system demonstrated modest precision (MSE = 0.147, 95% CI 0.065 to 0.228) and relatively little bias (ME = 0.090, 95% CI -0.025 to 0.205). Bland-Altman analysis also suggested good agreement at average INRs from 2.0 to 3.0. At average INR values >3.0, the POC system consistently overestimated INR. Values obtained with the POC system were significantly correlated with those obtained from the hospital laboratory (r = 0.77; p < 0.001). Similarly, mean +/- SD POC INR did not differ significantly from the laboratory-determined INR (2.45+/-0.59 vs. 2.37+/-0.48, respectively; p = 0.176). Regarding clinical accuracy, the values clinically agreed in 85.4% of the cases. CONCLUSIONS: The AvoSure PT Pro POC system appears to be useful for INR values within the 2.0-3.0 range, but values outside of this range should probably be confirmed with a standard laboratory method. PMID- 11895048 TI - Reimbursement claims analysis of outcomes with carvedilol and metoprolol. AB - OBJECTIVE: To compare resource use and costs in heart failure (HF) patients receiving metoprolol, a selective beta1-receptor blocker, with carvedilol, which blocks beta1-, beta2-, and alpha1-adrenergic receptors, by use of a retrospective reimbursement-claims analysis. METHODS: Resource use and cost data were extracted for patients diagnosed with HF and treated with carvedilol or metoprolol for 6 months after the initiation of the respective therapy, by use of claims submitted to 6 healthcare plans. A modified Charlson index was used to assess comorbidity. Stepwise logistic regression was used to measure the influence of treatment on hospitalization. RESULTS: Claims from 139 carvedilol and 106 metoprolol patients showed that carvedilol patients experienced significantly fewer total hospitalizations (36.0% vs. 62.3%, respectively; p < 0.001) and emergency department visits (23.7% vs. 42.5%, respectively; p = 0.002) and a trend for fewer HF-related (7.9% vs. 14.2%, respectively; NS) and cardiac-related hospitalizations (15.1% vs. 24.5%, respectively; NS). Treatment with carvedilol was associated with a significant decrease in the risk of any hospitalization (adjusted odds ratio 0.35, 95% CI 0.20 to 0.63; p <0.001). Higher pharmacy costs (mean $1677 vs. $1322; p <0.001) and lower total costs (mean $8100 vs. $14475; p = 0.025) were observed in carvedilol-treated compared with metoprolol-treated patients, respectively. CONCLUSIONS: Compared with metoprolol, the more comprehensive adrenergic blockade achieved with carvedilol may translate into greater clinical benefits in patients with HF. Despite higher pharmacy costs, lower total costs were observed in carvedilol-treated patients. PMID- 11895049 TI - Effect of colesevelam on lovastatin pharmacokinetics. AB - OBJECTIVE: To assess potential interactions of colesevelam hydrochloride and lovastatin in healthy volunteers when lovastatin alone was administered with dinner, both lovastatin and colesevelam were administered with dinner, and colesevelam was administered with dinner and lovastatin was administered 4 hours later with a snack. METHODS: A single-center, open-label, 3-period, crossover drug interaction study was performed with 22 healthy volunteers. Blood samples were collected at specified intervals before and after dosing, and plasma concentrations of lovastatin and lovastatin hydroxyacid were measured using a liquid chromatography/mass spectroscopy/mass spectroscopy method. RESULTS: Maximal concentration (Cmax), AUC from time 0 to the last time point measured (AUC0-t), and AUC0-infinity values for lovastatin were 102%, 94%, and 104%, and for lovastatin hydroxyacid were 102%, 91%, and 92%, respectively, of control values when colesevelam and lovastatin were coadministered with dinner. Administration of colesevelam with dinner and lovastatin 4 hours later with a snack resulted in a decreased Cmax and AUC0-t for lovastatin (63% and 37%, respectively; p < 0.05) and an increased Cmax and AUC0-t for lovastatin hydroxyacid (61% and 50%, respectively; p < 0.05), both compared with lovastatin alone administered with dinner. CONCLUSIONS: Colesevelam had no significant effect on lovastatin pharmacokinetics when coadministered with lovastatin at dinner. A split-dosing regimen resulted in alterations in pharmacokinetic parameters for lovastatin and lovastatin hydroxyacid that are likely due to known differences in the pharmacokinetics of lovastatin when administered to patients with meals or in a fasting state. PMID- 11895050 TI - Absorption of colesevelam hydrochloride in healthy volunteers. AB - OBJECTIVE: To assess whether colesevelam hydrochloride is absorbed in healthy volunteers. METHODS: A single-center, open-label, radiolabeled study was performed with 16 healthy volunteers. Subjects were administered non-radiolabeled colesevelam hydrochloride 1.9 g twice daily for 4 weeks, followed by a single dose of [14C]-colesevelam 2.4 g (480 pCi). These subjects continued to receive non-radioactive colesevelam 1.9 g twice daily for 4 days after administration of the radiolabeled dose. Blood, urine, and feces were collected immediately prior to administration of [14C]-colesevelam and at specified intervals after administration. The whole-blood equivalent concentration of colesevelam was calculated using data collected throughout the 96 hours following radiolabeled drug administration. The proportion of [14C]-colesevelam excreted through urine or feces was calculated based on the amount of radioactivity recovered up to 216 hours after the radiolabeled dose. RESULTS: The mean cumulative total recovery of [14C]-colesevelam in urine and feces was 0.05% and 74%, respectively. Excluding 2 subjects for whom cumulative recovery was <25%, the mean cumulative fecal recovery was 82%. The mean maximum whole-blood equivalent concentration of colesevelam was 0.165+/-0.10 microg equiv/g 72 hours after administration of [14C]-colesevelam, which was estimated to be 0.04% of the administered dose. All blood samples contained <4 times the number of background counts (dpm). CONCLUSIONS: The cumulative recovery data in urine and feces are consistent with the conclusion that colesevelam is not absorbed and is excreted entirely through the gastrointestinal system. PMID- 11895051 TI - Health survey data on potentially inappropriate geriatric drug use. AB - BACKGROUND: Studies have suggested that elderly patients do not always use medications appropriately. Investigations that have relied on prescription claim databases or clinical records focus on acquisition or prescription, and hypotheses must be made to assess actual consumption. Population survey data constitute an altemative way to study inappropriate use. OBJECTIVE: To estimate the prevalence of potentially inappropriate use of medications in elderly patients in Quebec based on self-reported use. METHODS: Using a cross-sectional, general population, health survey in which self-reported medication use in the 2 days prior to the survey was recorded, we estimated the prevalence of inappropriate medication use in elderly patients (> or =65 y old) who responded. Two sets of published criteria were used to define inappropriate use: one to assess use of inappropriate drugs, and another to assess concomitant duplications and potential interactions. RESULTS: Of the 3400 patients surveyed, 6.5% had used > or =1 inappropriate drugs, 2.5% had > or =1 occurrences of potentially inappropriate duplication of medications, and 2.7% had > or =1 potential medication interactions. Concomitant use of at least 2 benzodiazepines was reported by 8.5% of respondents using these drugs. Use of > or =1 long-acting benzodiazepines was reported by 4.2% of the sample. CONCLUSIONS: Population health surveys are a useful tool for detecting potentially inappropriate medication use in the elderly. In particular, the high prevalence of inappropriate use of benzodiazepines signals a need for improved detection and intervention in this group. PMID- 11895052 TI - Broth microdilution and E-test for determining fluoroquinolone activity against Streptococcus pneumoniae. AB - OBJECTIVE: To compare broth microdilution and E-test minimum inhibitory concentrations (MICs) of 4 fluoroquinolones against Streptococcus pneumoniae and to determine the effect of these in vitro MIC methods on the calculation of AUC00 24/MIC ratios. METHODS: Levofloxacin, gatifloxacin, moxifloxacin, and gemifloxacin MICs were determined by broth microdilution (incubated in air) and E test (incubated in CO2) for 100 clinical isolates of S. pneumoniae. MIC50, MIC90, and geometric mean MIC were calculated. Steady-state serum concentration-time profiles were simulated for once-daily, oral dosing of levofloxacin 500 mg, gatifloxacin 400 mg, moxifloxacin 400 mg, and gemifloxacin 320 mg. After correcting for protein binding, AUC0-24 of unbound drug was calculated for each regimen, and AUC0-24/MIC ratios were calculated using MIC data from both in vitro methods. Differences in MICs between methods were determined for each agent using the paired t-test (after logarithmic transformation of MICs) and the Wilcoxon signed-rank test. Differences in AUC0-24/MIC ratios were also determined using the paired t-test and the Wilcoxon signed-rank test. The level of significance for all analyses was p < 0.05. RESULTS: Broth microdilution and E-test MICs were within +/- 1 log2 dilution for 94%, 93%, 61%, and 35% of the isolates for levofloxacin, gatifloxacin, moxifloxacin, and gemifloxacin, respectively. Broth microdilution MICs were significantly lower than E-test MICs for all 4 agents (p < 0.001). However, a categorical change in susceptibility was seen for only 1 isolate with gatifloxacin and moxifloxacin (intermediate by broth microdilution, resistant by E-test). AUC0-24/MIC ratios were significantly higher for each regimen when MICs were determined by broth microdilution compared with E-test (p < 0.001). CONCLUSIONS: There is a significant difference in the activity of the newer fluoroquinolones against S. pneumoniae when MICs are determined by broth microdilution and E-test. When evaluating fluoroquinolone activity and pharmacodynamics against this organism, clinicians must be aware that MIC testing methodology may have a significant impact on the results. PMID- 11895053 TI - Stability of furosemide in human albumin solution. AB - OBJECTIVE: To determine the chemical stability of furosemide in human albumin solution over a 28-day period and to assess admixtures for microbiologic contamination. METHODS: Samples were prepared by mixing furosemide injectable solution and 25% human albumin solution in a 1:1 molar ratio. Six bulk containers were prepared and stored in the dark: 3 under refrigeration (approximately 4 degrees C) and 3 at room temperature (approximately 25 degrees C). Study samples were withdrawn from each bulk solution immediately after preparation and at predetermined intervals over the subsequent 28 days. Containers were observed for color change and precipitation against a light and dark background at each sampling interval. Total furosemide concentration was determined using HPLC. Additional vials were prepared and assessed for microbiologic growth at time points corresponding with chemical stability results. RESULTS: A mean of 94.5%+/ 1.33% of the initial furosemide concentration remained after 48 hours at room temperature. Under refrigeration, 100.6%+/-1.02% of the initial concentration remained at 14 days. Beyond these respective time points, <90% of the initial furosemide concentration remained. No bacterial or fungal growth was observed. CONCLUSIONS: When combined with 25% human albumin solution and stored under darkness, furosemide is chemically stable and free of microbiologic contamination for 48 hours at room temperature and 14 days under refrigeration. PMID- 11895054 TI - Efavirenz-induced skin eruption and successful desensitization. AB - OBJECTIVE: To report a patient with efavirenz-induced hypersensitivity syndrome reaction who was successfully desensitized to efavirenz. CASE SUMMARY: A 37-year old HIV-positive white man was placed on efavirenz, amprenavir, stavudine, lamivudine, and didanosine due to virologic failure with a previous regimen. Eight days into treatment, the patient developed a generalized rash and all medication was discontinued. Two weeks later, he was started on efavirenz, stavudine, didanosine, lamivudine, and lopinavir. The next morning, he awoke with red, itchy skin. All of the medications were discontinued. At the HIV Drug Safety Clinic, the patient was successfully restarted on amprenavir, stavudine, didanosine, lamivudine, and lopinavir. A 14-day desensitization to efavirenz was also undertaken; on day 12 of the desensitization, he once again developed a rash. He was treated symptomatically, and the desensitization protocol was extended. Sixteen months following the successful desensitization, he is tolerating full-dose efavirenz in combination with the other antiretroviral agents. DISCUSSION: The incidence of efavirenz-induced hypersensitivity ranges between 10% and 34%. Generally, an erythematous, maculopapular rash with or without fever develops 1-3 weeks after the initiation of therapy. In many patients without systemic manifestations, an attempt should be made to continue therapy and treat the rash symptomatically. If this fails, desensitization with the implicated drug can be tried. CONCLUSIONS: A history of antiretroviral induced hypersensitivity reactions often limits the choices of medications that can be used in subsequent treatment regimens. Desensitization may allow for the continued use of previously restricted medications. PMID- 11895055 TI - Gabapentin for hot flashes in prostate cancer. AB - OBJECTIVE: To report a case of successful treatment of refractory hot flashes with gabapentin in a patient with prostate cancer who was receiving combination antiandrogen and gonadotropin hormone-releasing hormone (GnRH) analog therapy. CASE SUMMARY: A 70-year-old white man with a history of advancing prostate cancer experienced disabling hot flashes from combination therapy with the antiandrogen bicalutamide and the GnRH analog goserelin acetate. He failed to respond to clonidine 0.1 mg twice daily, megestrol acetate 40 mg/d, diethylstilbestrol 1 mg/d, and venlafaxine 25 mg twice daily. The patient was then treated with gabapentin 600 mg once daily, at which time he experienced near-complete resolution of his symptoms. DISCUSSION: This case supports a previous report of the marked improvement in severity and duration of hot flashes associated with antiandrogen or GnRH analog therapy in prostate cancer. The mechanism by which gabapentin reduces hot flashes is unknown. CONCLUSIONS: Hot flashes resulting from antiandrogen or GnRH analog therapy are often difficult to treat and leave many patients disabled. Gabapentin has been shown to markedly reduce the severity, frequency, and duration of these hot flashes. Controlled trials are necessary to evaluate gabapentin against other therapeutic modalities. PMID- 11895057 TI - Neurotoxic syndrome associated with risperidone and fluvoxamine. AB - OBJECTIVE: To report a case of a neurotoxic syndrome in a patient undergoing concomitant treatment with risperidone and fluvoxamine. CASE REPORT: A 24-year old African American woman hospitalized for psychosis was unresponsive to risperidone. Because of obsessive symptoms, low doses of fluvoxamine were added to her treatment regimen. Within 2 days, she developed confusion, diaphoresis, diarrhea, hyperreflexia, and myoclonus, which then progressed to rigidity, fever, and unresponsiveness, requiring endotracheal intubation. Symptoms resolved over 10 days with discontinuation of medication, hydration, and bromocriptine 5 mg 3 times daily. Ultimately, she was treated with olanzapine and fluvoxamine without adverse effects. DISCUSSION: This represents the first reported case of a neurotoxic syndrome secondary to treatment with risperidone and fluvoxamine. Differential diagnosis between neuroleptic malignant syndrome (NMS) and serotonin syndrome (SS) could not be accurately determined because of the overlap of signs and symptoms of both syndromes. NMS and SS may represent different aspects of a more generalized neurotoxic syndrome. This could be an important consideration in formulating treatment for neurotoxic syndromes. CONCLUSIONS: Clinicians should be aware of potentially serious adverse reactions that may occur during concomitant treatment with antipsychotics and selective serotonin-reuptake inhibitors. PMID- 11895056 TI - New-onset seizure associated with quetiapine and olanzapine. AB - OBJECTIVE: To report a case involving a witnessed seizure in a patient receiving concurrent olanzapine and quetiapine. CASE SUMMARY: A 27-year-old white woman was observed to have a seizure while receiving a stable dosage of olanzapine 15 mg/d, with the addition of quetiapine 100 mg in the evening 1 day before the occurrence of the seizure. There were no known risk factors for epilepsy. DISCUSSION: This case reports a new-onset seizure in the context of concurrent olanzapine and quetiapine use. Interpretation is complicated by recent discontinuation of low dose clonazepam. CONCLUSIONS: While uncommon, seizures can occur with non clozapine atypical antipsychotics. Caution is indicated when using these drugs with other agents that may lower the seizure threshold. PMID- 11895058 TI - Breast enlargement induced by D-penicillamine. AB - OBJECTIVE: To report a patient with reversible breast enlargement due to D penicillamine treatment for scleroderma. CASE SUMMARY: A 55-year-old white woman developed enlarged breasts during D-penicillamine therapy for scleroderma. The enlargement regressed following discontinuation of the drug, and planned breast reduction surgery was avoided. DISCUSSION: D-Penicillamine has been used in the treatment of various diseases including scleroderma. Breast enlargement has been previously described as a rare adverse effect of this drug. The enlargement subsided following discontinuation of the medication. CONCLUSIONS: Attention is called to this rare adverse effect in an effort to avoid unnecessary diagnostic or therapeutic procedures, including surgery. PMID- 11895059 TI - Ototoxicity induced by gentamicin and furosemide. AB - OBJECTIVE: To present a case of ototoxicity induced by furosemide and once-daily gentamicin therapy. CASE SUMMARY: A 60-year-old white woman presented to the hospital with community-acquired pneumonia and urinary tract infection. The antibiotic regimen included gentamicin and, after 5 doses, the patient reported profound bilateral hearing loss. A Pure Tone Audiogram suggested moderate to moderately severe sensorineural hearing loss bilaterally. The only risk factors present included her age, elevated temperature, and the use of furosemide. DISCUSSION: Several risk factors may predispose a patient to developing aminoglycoside ototoxicity: the 1555 chromosomal mutation, preexisting disorders of hearing and balance, hypovolemia, bacteremia, liver and renal dysfunction, and the simultaneous administration of other ototoxic medications. The cumulative dose and duration of aminoglycoside therapy are more important than serum concentrations. Administration of an aminoglycoside followed by furosemide may increase the risk of ototoxicity. The aminoglycoside interacts with the cell membranes in the inner ear, increasing their permeability. This theoretically allows the loop diuretic to penetrate into the cells in higher concentrations, causing more severe damage. CONCLUSIONS: Auditory toxicity occurred after only 5 days of gentamicin therapy and 1 dose of furosemide. An aminoglycoside followed by furosemide may increase the risk for ototoxicity. Clinicians need to be aware of the synergistic potential of ototoxic medications. PMID- 11895060 TI - Telithromycin: the first of the ketolides. AB - OBJECTIVE: To review the chemistry, spectrum of activity, pharmacology, clinical efficacy, and safety of telithromycin. DATA SOURCES: A MEDLINE search from 1966 to December 2000 was performed via OVID and PubMed using the following search terms: HMR 3647, HMR3647, Ketek, RU 66647, and telithromycin. An extensive review of retrieved literature, abstracts from international scientific conferences, and minutes from regulatory authority meetings was also performed. DATA EXTRACTION: Medicinal chemistry, in vitro, animal, and human trials were reviewed for information on the antimicrobial activity, clinical efficacy, pharmacology, and safety of telithromycin. DATA SYNTHESIS: Several chemical modifications to the macrolide structure have led to the development of telithromycin, the first ketolide antimicrobial that demonstrates improved activity against penicillin- and macrolide/azalide-resistant Streptococcus pneumoniae due to its unique binding to the ribosomal target site. Although telithromycin may be useful in the treatment of community-acquired respiratory tract infections due to its activity against common typical and atypical pathogens, questions concerning its reliable activity against Haemophilus influenzae need to be addressed. Telithromycin's pharmacokinetics permit once-daily dosing for abbreviated periods and good distribution into lung tissue and phagocytic cells. Clinical and bacteriologic cure rates have been similar to those of comparator agents in human efficacy trials; however, the incidence of adverse gastrointestinal events were generally higher with telithromycin patients. Like other macrolides and many newer fluoroquinolones, telithromycin's ability to prolong the QTc interval is a potential safety issue, especially in elderly patients with predisposing conditions or those who are concurrently receiving drugs that are substrates for CYP2D6 and 3A4. Liver function test elevations demonstrated during clinical trials, although not overtly severe, may warrant monitoring in some patients taking multiple hepatically metabolized/cleared agents. CONCLUSIONS: Telithromycin offers potential advantages over traditional macrolides/azalides for community-acquired respiratory tract infections caused by macrolide-resistant pathogens. Further studies are needed to elucidate its clinical efficacy against H. influenzae, potential drug interactions, and safety in various subpopulations. PMID- 11895061 TI - Postfertilization effect of hormonal emergency contraception. AB - OBJECTIVE: To assess the possibility of a postfertilization effect in regard to the most common types of hormonal emergency contraception (EC) used in the US and to explore the ethical impact of this possibility. DATA SOURCES AND STUDY SELECTION: A MEDLINE search (1966-November 2001) was done to identify all pertinent English-language journal articles. A review of reference sections of the major review articles was performed to identify additional articles. Search terms included emergency contraception, postcoital contraception, postfertilization effect, Yuzpe regimen, levonorgestrel, mechanism of action, Plan B. DATA SYNTHESIS: The 2 most common types of hormonal EC used in the US are the Yuzpe regimen (high-dose ethinyl estradiol with high-dose levonorgestrel) and Plan B (high-dose levonorgestrel alone). Although both methods sometimes stop ovulation, they may also act by reducing the probability of implantation, due to their adverse effect on the endometrium (a postfertilization effect). The available evidence for a postfertilization effect is moderately strong, whether hormonal EC is used in the preovulatory, ovulatory, or postovulatory phase of the menstrual cycle. CONCLUSIONS: Based on the present theoretical and empirical evidence, both the Yuzpe regimen and Plan B likely act at times by causing a postfertilization effect, regardless of when in the menstrual cycle they are used. These findings have potential implications in such areas as informed consent, emergency department protocols, and conscience clauses. PMID- 11895062 TI - Role of race in the pharmacotherapy of heart failure. AB - OBJECTIVE: To review the literature assessing the differences in response to angiotensin-converting enzyme (ACE) inhibitors and beta-blockers in black patients compared with the response in non-black patients in the management of systolic heart failure. DATA SOURCES: A MEDLINE search (January 1966-May 2001) was performed using heart failure, blacks, Negroid race, adrenergic beta antagonists, and angiotensin-converting enzyme inhibitors as key words. English language articles were identified. Additional pertinent articles were identified from review of the references of these articles. STUDY SELECTION AND DATA EXTRACTION: All identified references were reviewed. All articles deemed relevant to the subject of this article were included. DATA SYNTHESIS: It has been suggested that the antihypertensive effect of ACE inhibitors and beta-blockers may be less in black patients than in other racial groups. Retrospective reanalyses of major heart failure trials have suggested that black patients may not realize a significant benefit in morbidity or mortality when heart failure is managed with ACE inhibitors or beta-blockers. It has also been suggested that black patients may respond more favorably than non-black patients to the combination of hydralazine and isosorbide dinitrate. CONCLUSIONS: Published reanalyses of ACE inhibitor and beta-blocker trials in heart failure provide weak data to support a lack of benefit in black patients. The published literature on this topic is limited by its retrospective nature. Firm conclusions regarding the influence of race on effectiveness of ACE inhibitors and beta-blockers cannot be made until prospective trials, with planned analysis of the effect of race, have been performed. PMID- 11895063 TI - Surrogate end points in heart failure. AB - OBJECTIVE: To describe the potential utility of a true surrogate marker of heart failure outcomes, historically investigate the validity of surrogates most commonly evaluated in heart failure, and identify specific end points offering the most potential for future use. DATA SOURCES: A MEDLINE search (1966-June 2001) was completed to identify relevant literature. Additional references were also retrieved from selected articles. Search terms included b-type natriuretic peptide, cardiac remodeling, end-diastolic volume, heart failure, and surrogate end points. DATA SYNTHESIS: By definition, true surrogate end points must predict outcomes associated with disease progression and response to therapy. A validated surrogate measure of mortality would render significant utility in both heart failure drug development and clinical practice. Improvements in traditional functional markers of heart failure, including ejection fraction and exercise capacity, have produced inconsistent results in regard to mortality in a number of clinical trials. Numerous measures of cardiac remodeling and neurohormonal activation, however, have proven to be reliable predictors of disease progression and therapeutic response. These findings have contributed significantly to the current understanding of heart failure pathophysiology and approach to treatment. Specifically, measures such as left-ventricular end-diastolic volume (LVEDV) and, potentially, b-type natriuretic peptide (BNP) concentrations may correlate with disease progression and parallel the mortality reductions observed with angiotensin-converting enzyme inhibitor and beta-blocker therapy. CONCLUSIONS: Currently, LVEDV and plasma BNP offer the greatest potential as surrogate end points in heart failure. Further investigation of these measures is required before routine implementation in drug development and clinical practice can be justified. PMID- 11895064 TI - Treatment of heparin-induced thrombocytopenia. AB - OBJECTIVE: To describe heparin-induced thrombocytopenia (HIT or HIT-2), an immune mediated adverse reaction to heparin or low-molecular-weight heparin. Available treatment options and considerations in developing a therapy approach are discussed. DATA SOURCES: A search of the National Library of Medicine (1992-June 2001) was done to identify pertinent literature. Additional references were reviewed from selected articles. STUDY SELECTION: Articles related to laboratory recognition and treatment options of HIT, including the use of agents in selected clinical conditions, were reviewed and included. CONCLUSIONS: HIT is a rare but potentially severe adverse reaction to heparin that was, until recently, poorly understood and had limited treatment options. Recent advances describing the recognition and clinical manifestations of immune-mediated HIT, including recently available antithrombotic treatment options, have dramatically changed outcomes for patients having this syndrome. PMID- 11895065 TI - Prostaglandin analog treatment of glaucoma and ocular hypertension. AB - OBJECTIVE: To review available data related to the use of prostaglandin analogs (bimatoprost, latanoprost, travoprost, unoprostone) in the management of ocular hypertension and open-angle glaucoma. DATA SOURCES: Primary and review articles were identified from a MEDLINE search (1966-May 2001) and requested information from product manufacturers. STUDY SELECTION AND DATA EXTRACTION: All available information, including that published in articles and abstracts, which was deemed relevant was included in this review. Limited data have been published to date. DATA SYNTHESIS: The prostaglandin analogs appear to be effective, well-tolerated agents for the reduction of intraocular pressure (IOP) in patients with primary open-angle glaucoma and ocular hypertension. This drug class offers an alternative for patients who do not achieve control with another topical antiglaucoma agent or for those with a contraindication to first-line therapy with beta-adrenergic antagonists. Based on preliminary clinical data, bimatoprost, latanoprost, and travoprost appear to be at least as effective as timolol, while the effectiveness of unoprostone is similar or slightly less. Prostaglandin analogs may be used in conjunction with other antiglaucoma medications, although further studies must establish the optimal combination. Whether clinical experience will yield outcomes in favor of one of the prostaglandin analogs remains to be determined. Patients should be educated on adverse events associated with prostaglandin analogs, particularly the potential for changes in the pigmentation of the iris and eyelashes. CONCLUSIONS: Bimatoprost, latanoprost, and travoprost appear to be equivalent to the current standard of therapy in the topical treatment of elevated IOP. Further clinical data published in article versus abstract format is required to better assess potential differences among these 3 agents. PMID- 11895066 TI - Risedronate for the prevention and treatment of corticosteroid-induced osteoporosis. AB - OBJECTIVE: To evaluate the role of risedronate in corticosteroid-induced osteoporosis. DATA SOURCES: Clinical literature was accessed through MEDLINE (1966-February 2001). Key search terms included risedronate, corticosteroid, osteoporosis, and bisphosphonate. DATA SYNTHESIS: Corticosteroid-induced osteoporosis (CIO) is clinically challenging and can lead to fractures. Risedronate, an oral bisphosphonate, has been studied for use in CIO. Trials focusing on the use of risedronate in these patients were reviewed. CONCLUSIONS: Risedronate 5 mg/d increased bone mineral density at lumbar, femoral neck, and trochanter skeletal sites in patients recently initiated on or receiving long term corticosteroid therapy. Further investigation is needed to determine risedronate's effects on fracture prevention. The drug was well tolerated. PMID- 11895067 TI - Granulocyte macrophage-colony stimulating factor and oral mucositis. AB - OBJECTIVE: To evaluate the effect of granulocyte macrophage-colony stimulating factor (GM-CSF) in patients with oral mucositis. DATA SOURCES: Literature was accessed through MEDLINE (1966-June 2000) and bibliographic searches. DATA SYNTHESIS: Published literature assessing the use of GM-CSF to treat or prevent oral mucositis was analyzed. CONCLUSIONS: Reports of GM-CSF improving mucositis have been published; however, because of limitations in these reports, insufficient evidence confirms the benefits of GM-CSF. These preliminary studies provide a rationale to conduct well-designed, double-blind, randomized trials to evaluate the safety and efficacy of GM-CSF in the treatment and/or prevention of oral mucositis induced by chemotherapy and/or radiotherapy. PMID- 11895068 TI - Thalidomide use in pediatric patients. AB - OBJECTIVE: To provide a detailed overview of thalidomide use in pediatric patients. DATA SOURCES: English-language articles were identified through a MEDLINE search (1966-February 2001); key terms included thalidomide, child, graft versus-host disease, cancer, HIV, Crohn's disease, Behcet's disease, and lupus erythematosus. References cited in those articles were also evaluated. DATA SYNTHESIS: Thalidomide appears to be effective in patients with chronic, not acute, graft-versus-host disease (GVHD) and in healing aphthous ulcers in patients with HIV infection. Limited case reports suggest efficacy of thalidomide in the treatment of cutaneous manifestations of Behcet's disease, Crohn's disease, and lupus in children; however, the recurrence of disease is almost universal on drug discontinuation. CONCLUSIONS: Thalidomide should be used as a last resort when all other therapies fail, preferably in male or prepubescent female patients. PMID- 11895069 TI - Acyclovir-induced crystalluria. PMID- 11895070 TI - Pharmacist's role in health care. PMID- 11895071 TI - Pharmacodynamics in the evaluation of drug regimens. AB - We believe, especially in an era of increasing antimicrobial resistance, that the integration of PK/PD models and stochastic techniques provides a powerful new tool to clinicians, researchers, and regulatory agencies alike. By using techniques such as these, we can begin to level the playing field so that PK/PD measures for therapeutic agents can be more accurately compared with greater certainty, and not be limited to discussions of susceptibility method variation or comparisons of MIC results. PMID- 11895072 TI - Cardiac arrhythmia in a newborn infant associated with fluoxetine use during pregnancy. PMID- 11895073 TI - Comment: prophylaxis for RSV hospitalization may not be cost-saving. PMID- 11895074 TI - Amino acids, then and now--a reflection on Sir Hans Krebs' contribution to nitrogen metabolism. AB - H. A. Krebs made an enormous contribution to our knowledge of amino acid metabolism, beginning with his studies on proteolysis in the early 1930s, progressing through his work on urea synthesis to an extensive series of papers on deamination and, then, to work on gluconeogenesis from amino acids. This paper addresses three of Krebs' early contributions-urea synthesis, glutamine metabolism, and D-amino acid oxidase-and relates them to our modern understanding of amino acid metabolism. PMID- 11895075 TI - Exploiting the versatility of human cytochrome P450 enzymes: the promise of blue roses from biotechnology. AB - The cytochrome P450 (P450) enzymes involved in drug metabolism are among the most versatile biological catalysts known. A small number of discrete forms of human P450 are capable of catalyzing the monooxygenation of a practically unlimited variety of xenobiotic substrates, with each enzyme showing a more or less wide and overlapping substrate range. This versatility makes P450s ideally suited as starting materials for engineering designer catalysts for industrial applications. In the course of heterologous expression of P450s in bacteria, we observed the unexpected formation of blue pigments. Although this was initially assumed to be an artifact, subsequent work led to the discovery of a new function of P450s in intermediary metabolism and toxicology, new screens for protein engineering, and potential applications in the dye and horticulture industries. PMID- 11895076 TI - Assessing skeletal muscle glucose metabolism with positron emission tomography. AB - Insulin has a marked effect to stimulate the transport and metabolism of glucose in skeletal muscle in healthy individuals, whereas an impaired response, termed insulin resistance, is a major risk factor for diabetes mellitus and other metabolic diseases. Studies of the molecular physiology of insulin action in skeletal muscle indicate that a principal loci of control resides within the proximal steps of glucose transport and phosphorylation. Deoxyglucose, the metabolism of which is limited to these proximal steps, is widely used for in vitro studies of insulin action on glucose transport. The technologies of PET imaging provide a unique opportunity to carry out similar studies in vivo in human skeletal muscle. In this instance, a short-lived positron labeled tracer, [18F] FDG, can be given at sufficiently high specific activity to image not only glucose uptake, but by dynamic PET imaging, by monitoring the time course of [18F] FDG tissue activity, data can be generated to examine the kinetics of glucose transport and phosphorylation. The experimental procedures of this approach, including an overview of the mathematical modeling, are described in this review, along with some of the key findings of the initial applications of PET for the study of glucose metabolism in human skeletal muscle. PMID- 11895077 TI - Novel approaches to tackling malarial drug resistance using yeast. AB - Yeasts have a justified reputation as one of the world's most versatile organisms. Baker's yeast continues to live up to this recognition by joining the war against malaria. Yeast can now be used to study antifolate drug resistance patterns that depend on the dihydrofolate reductase enzyme (DHFR) from the malaria parasite. PMID- 11895078 TI - Multidimensional NMR methods for protein structure determination. AB - Structural studies of proteins are critical for understanding biological processes at the molecular level. Nuclear magnetic resonance (NMR) spectroscopy is a powerful technique for obtaining structural and dynamic information on proteins and protein-ligand complexes. In the present review, methodologies for NMR structure determination of proteins and macromolecular complexes are described. In addition, a number of recent advances that reduce the molecular weight limitations previously imposed on NMR studies of biomolecules are discussed, highlighting applications of these technologies to protein systems studied in our laboratories. PMID- 11895079 TI - Hsp27: novel regulator of intracellular redox state. AB - Small stress proteins are molecular chaperones that modulate the ability of cells to respond to several types of injuries. In this regard, a protection generated by the expression of mammalian small stress proteins against the cell death induced by oxidative stress has been described. This review summarizes the current knowledge of the protective mechanism generated by the expression of the major mammalian small stress protein Hsp27 in cells exposed to oxidative stress. A possible role of this chaperone protein in the presentation of oxidized proteins to the proteasome degradation machinery is proposed. PMID- 11895080 TI - Mutational analysis of the three conserved valine residues of insulin and a proposal of "isosteric residue". AB - To further understand the role of the three conserved Val residues in insulin, B12Val, B18Val, and A3Val, five insulin mutants-[A3Ser]insulin, [B12Thr]insulin, (desB30)[B12Ser]insulin, [B18Thr] insulin, and [B18Leu]insulin--were obtained by means of site-directed mutagenesis and their receptor-binding activities as well as in vivo biological potencies were measured. The two B18 mutants, [B18Thr]insulin and [B18Leu]insulin, both retained relatively high receptor binding activities (70% and 30% of native porcine insulin, respectively) as well as relatively high in vivo biological potencies. The receptor-binding activities of [B12Thr]insulin and (desB30)[B12Ser]insulin were 5.1% and 0.2%, respectively. However the in vivo biological potency of [B12Thr]insulin was still about 50% of native insulin, whereas that of (desB30)[B12Ser]insulin decreased drastically. The [A3Ser]insulin retained 1.4% of the receptor-binding activity and low in vivo biological potency. These results, together with previous reports showed that when the three conserved Val residues were replaced by residues containing a beta branched side-chain, such as Thr or Ile, the insulin mutants retained higher biological activities than those mutants replaced by other residues. Here we propose that Val, Thr, and Ile are "isosteric residues' because they all contain a beta-branched side-chain. This proposal may have perhaps general significance in protein design and protein engineering. PMID- 11895081 TI - Pressure overload induces heterologous expression of the atrial natriuretic factor (ANF) gene. AB - Several investigations have demonstrated the regional heterogeneity of myocardial phenotype, and hypertrophy may also induce regionally disparate changes. We have utilized the direct DNA injection technique to study regional variations in overload-induced ANF expression. Pressure overload was induced by stenosis of the ascending aorta in canines. ANF promoter reporters were injected into the left ventricle; in different regions including the base, the midwall region, and the apex. Injections were made at different depths to include the epicardial and endocardial layers. The animals were sacrificed 7 days following surgery and the left ventricle harvested for tissue analysis. Under normotensive conditions, ANF reporter expression was similar throughout the heart. PO increased ANF expression and the increases were greater in the endocardium than in the epicardium. PO also significantly increased expression in the midwall and base regions, but not in the apex. It is unknown from these experiments, whether the greater increases in midwall expression are a function of greater wall stress, metabolic demand, or phenotypic differences in the midwall myocytes. These findings do indicate that regional differences in overload-induced changes in gene expression are evident and may be functionally important in determining myocardial response to increased functional demand. PMID- 11895082 TI - Transfer of cationic antibacterial agents berberine, palmatine, and benzalkonium through bimolecular planar phospholipid film and Staphylococcus aureus membrane. AB - Some organic cations are known to be electrophoretically imported into bacterial cells and actively extruded from these cells by multidrug resistance (MDR) pumps. We have studied penetration of plant antimicrobial agents berberine and palmatine and synthetic antiseptic benzalkonium chloride through black planar phospholipid membrane (BLM) and membrane of Staphylococcus aureus cells. Gradients of these cations across BLM generated an electric potential difference. Penetrating anion tetraphenyl borate and phloretin (a plant substance decreasing membrane dipole potential) stimulated this effect. Under optimal conditions, the magnitude of the electric potential was close to theoretical, that is, 60 mV/10-fold cation gradient. Berberine accumulated in S. aureus cells as shown by direct measurement of berberine with a berberine-sensitive electrode. The berberine accumulation was prevented by protonophore CCCP and was stimulated by mutation in the MDR pump NorA. It is concluded that the plant alkaloids and benzalkonium are penetrating cations and substrates of an MDR pump. PMID- 11895084 TI - A listeriosis patient infected with two different Listeria monocytogenes strains. AB - Normally, only one isolate of Listeria monocytogenes from a case of listeriosis is subjected to characterization. Here we show that two isolates from different sites of the body were not the same strain. Such a phenomenon may not have any clinical relevance, although it may confuse the epidemiologist trying to match infection source with infection target. PMID- 11895083 TI - An outbreak of multidrug-resistant Salmonella enterica subsp. enterica serotype Typhimurium, DT104L linked to dried anchovy in Singapore. AB - Multidrug-resistant Salmonella enterica subsp. enterica serotype Typhimurium DT104L was first reported in Singapore from mid-July to mid-October 2000. Salmonella strains isolated from clinical laboratories were submitted to a reference laboratory for serotyping, phage-typing and pulsed-field gel electrophoresis (PFGE) using XbaI restriction endonuclease. An epidemiological investigation was conducted to determine the source of infection and mode of transmission using a structured questionnaire. A total of 33 cases involving mainly infants and toddlers were detected in the 3-month long outbreak. The outbreak strain was of the R-type ACGSTSu, i.e. resistant to ampicillin, chloramphenicol, gentamicin, streptomycin, tetracycline and sulphonamide. PFGE showed all isolates had an indistinguishable pattern, indicating a common source of infection. Consumption of imported dried anchovy was found to be the vehicle of transmission after adjusting for all confounding variables in the case-control study using stepwise logistic regression (OR 25.6; 95% CI 3.9-167.9; P = 0.001). Imported dried seafood should be properly processed, packed, labelled, and thoroughly cooked to prevent transmission of multidrug-resistant S. Typhimurium. PMID- 11895085 TI - Prevalence of Leptospira spp. in various species of small mammals caught in an inner-city area in Switzerland. AB - In order to establish the leptospira carrier rate of small animals in an urban environment, small rodents and shrews were captured in the city of Zurich, Switzerland. Kidney specimens of 190 animals were examined using a leptospira specific PCR assay. Leptospiral DNA was amplified in kidneys of 12.6% of the animals. PMID- 11895086 TI - Molecular subtyping of Staphylococcus aureus from an outbreak associated with a food handler. AB - On 6 May 2000, a staphylococcal food poisoning outbreak occurred at a high school, affecting 10 of the 356 students who attended the breakfast. Twenty-seven Staphylococcus aureus isolates, producing enterotoxin A (SEA), SEB-, or non-SEA E, were recovered from 7 patients, 2 food handlers and left-overs. To investigate the outbreak, we genotyped the isolates by using pulsed-field gel electrophoresis (PFGE) and three PCR-based techniques: inter-IS256 PCR typing, protein A gene (spa) typing, and coagulase gene restriction profile (CRP) analysis. Our results show that PFGE was the most discriminatory technique, whereas the three PCR-based techniques were insufficient in the discriminatory power to distinguish the S. aureus isolates from the outbreak. Based on the enterotoxin-producing types and the results of genotyping, three distinct types of strains (A1111, B2221 and N3221) were designated. Both the A1111 and B2221 strains were found in the specimens from the patients and a hand lesion of a food handler, suggesting that the source of contamination for the outbreak was most likely originated from a food handler. PMID- 11895087 TI - Non-invasive pneumococcal disease and antimicrobial resistance: vaccine implications. AB - We reviewed laboratory data on non-invasive pneumococcal isolates reported from all diagnostic laboratories in Scotland during the period 1988-99. Of 4491 isolates from hospitalized patients, 654 (64.7%) were from sputum, 79 (7.8%) from the nasopharynx and 278 (27.5%) from other superficial sites. The serogroups included in the 23-valent polysaccharide vaccine caused 96.9% of all non-invasive disease in all age groups. The 7-, 9-, and 11-valent conjugated vaccine serogroups were responsible for 87-94%, 85-93%, 74-81% and 75-84% of non-invasive disease respectively in age groups < 2 years, < or = 5 years, > or = 65 years and all ages. The coverage of non-susceptible penicillin and erythromycin non invasive isolates was > 99% and > 95% with the 23-valent polysaccharide and 7-11 valent conjugate vaccines respectively. The eight most common serogroups were 23, 9, 6, 19, 14, 3, 15 and 11 (in descending order). The serogroups associated with antimicrobial resistance in non-invasive disease were similar to those found in invasive disease. The finding of a similar serogroup distribution in both invasive and non-invasive disease (regardless of the site of clinical isolate), is consistent with serogroups colonizing non-sterile sites and having the potential to invade. The availability of conjugated vaccines reinforces the importance of systematic surveillance to determine accurately and regularly the coverage of pneumococcal serogroups and types causing both invasive and non invasive disease. PMID- 11895088 TI - Changing epidemiology of human leptospirosis in New Zealand. AB - The objective was to describe the current epidemiology and trends in New Zealand human leptospirosis, using descriptive epidemiology of laboratory surveillance and disease notification data, 1990-8. The annual incidence of human leptospirosis in New Zealand 1990-8 was 44 per 100,000. Incidence was highest among meat processing workers (163.5/100,000), livestock farm workers (91.7), and forestry-related workers (24.1). The most commonly detected serovars were Leptospira borgpetersenii serovar (sv.) hardjo (hardjobovis) (46.1%), L. interrogans sv. pomona (24.4%) and L. borgpetersenii sv. ballum (11.9%). The annual incidence of leptospirosis declined from 5.7/100,000 in 1990-2 to 2.9/100,000 in 1996-8. Incidence of L. borgpetersenii sv. hardjo and L. interrogans sv. pomona infection declined, while incidence of L. borgpetersenii sv. ballum infection increased. The incidence of human leptospirosis in New Zealand remains high for a temperate developed country. Increasing L. borgpetersenii sv. ballum case numbers suggest changing transmission patterns via direct or indirect exposure to contaminated surface water. Targeted and evaluated disease control programmes should be renewed. PMID- 11895089 TI - Predictors of tuberculin reactivity among prospective Vietnamese migrants: the effect of smoking. AB - We investigated the prevalence and predictors of positive tuberculin skin test (TST) results among prospective Vietnamese migrants. We interviewed and medically screened 1395 Vietnamese people aged over 15 years who had applied to migrate to Australia. Approximately 44% of applicants had an induration of 10 mm or more, and 18.6% had an induration of 15 mm or more. A positive tuberculin skin test at 5 mm, 10 mm and 15 mm of induration cut-points was significantly associated with age (OR 1.01-1.02 per year) and duration of smoking (OR 1.03-1.12 per year). Smoking appears to be an important factor associated with increased susceptibility to mycobacterial infection. It is not yet clear whether the increased tuberculin reactivity associated with smoking reflects an increased risk of tuberculosis among these migrants. PMID- 11895090 TI - Protecting the vaccinating population in the face of a measles epidemic: assessing the impact of adjusted vaccination schedules. AB - We investigated which vaccination schedule gives best protection to the vaccinating population, in case there is a measles epidemic in an area with low vaccine coverage. We considered combinations of an early measles vaccination (none, at 6 months or at 9 months), a measles-mumps-rubella (MMR) vaccination around the first birthday (at either 11 or 14 months), and MMR vaccination at an older age (at either 4 or 9 years). The different estimates on measures of protection (percentage of susceptibles, number of reported cases in an epidemic year, percentage of lifetime spent susceptible) relied on a mathematical model of decline of maternal antibody levels with age, and the impact of that antibody level on seroconversion and immunity. Model parameters were estimated from a Dutch population-based serological survey on measles antibodies. Different measures of protection favoured different vaccination schedules, but dropping the age of second MMR vaccination prevents considerably more cases than an extra early measles vaccination or dropping the age of first MMR vaccination. PMID- 11895091 TI - Seroprevalence of parvovirus B19 in urban Chilean children and young adults, 1990 and 1996. AB - An immunofluorescence test for detecting parvovirus B19 IgG was developed by infecting insect cells with recombinant baculovirus expressing the capsid protein VPI. The test was used to study the prevalence of antibodies in 725 healthy children and young adults living in Santiago, Chile. In total, 248 sera were taken in 1990 and 477 in 1996. The seroprevalence was low in children less than 5 years old (3% in 1990 and 21% in 1996). It rose during school age to a prevalence around 50%, reaching 60% in young adults. No differences were found between genders. There was a statistically significant higher seroprevalence in the low socioeconomic status group in 1990 samples, but this was not observed in 1996. The higher prevalence observed in children less than 5 years of age in 1996 compared with 1990 could be explained by the occurrence of intervening epidemics of parvovirus B19 infection. PMID- 11895092 TI - Tattooing and risk for transfusion-transmitted diseases: the role of the type, number and design of the tattoos, and the conditions in which they were performed. AB - Tattoos have been shown to be associated with transfusion-transmitted diseases (TTDs), particularly hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Very little is known about the association between different categories of tattoos and TTDs. In a cross-sectional study in Brazil, we studied 182 individuals with tattoos and assessed the odds of testing positive for a TTD according to tattoo type, number, design and performance conditions. Major findings were significant associations between an increasing number of tattoos and HBV infection (odds ratio (OR) of 2.04 for two tattoos and 3.48 for > or = 3 tattoos), having a non-professional tattoo and testing positive for at least one TTD (OR = 3.25), and having > or = 3 tattoos and testing positive for at least one TTD (OR = 2.98). We suggest that non-professional tattoos and number of tattoos should be assessed as potential deferral criteria in screening blood donors. PMID- 11895093 TI - Increase in meningococcal disease associated with the emergence of a novel ST-11 variant of serogroup C Neisseria meningitidis in Victoria, Australia, 1999-2000. AB - In the years 1999-2000, there was an increase in the incidence of meningococcal disease in Victoria, largely caused by Neisseria meningitidis serogroup C. This change was associated with a shift in age distribution of cases, with relatively more disease appearing in the 15-29 year age group, and with 40/58 serogroup C isolates in 2000 exhibiting a new macrorestriction pattern (pattern A). Thirty four of 52 pattern A isolates tested displayed the novel phenotype C:2a:P1.4, and were consistently porA VR type P1.7-2,4 by DNA sequencing. Nine of 10 representative pattern A isolates analysed displayed a housekeeping gene allele profile (ST-11) that is characteristic of the electrophoretic type (ET)-15 variant that has caused outbreaks in Canada, the Czech Republic and Greece. Meningococci belonging to the ST-11 complex that were isolated in Victoria prior to 1999 did not display either restriction pattern A or PorA VR type P1.7-2,4. PMID- 11895094 TI - Risk factors in HIV-associated diarrhoeal disease: the role of drinking water, medication and immune status. AB - In a cross-sectional survey of 226 HIV-infected men, we examined the occurrence of diarrhoea and its relationship to drinking water consumption patterns, risk behaviours, immune status and medication use. Diarrhoea was reported by 47% of the respondents. Neither drinking boiled nor filtered water was significantly associated with diarrhoea (OR = 0.5 [0.2, 1.6], 1.2 [0.6, 2.5] respectively), whereas those that drank bottled water were at risk for diarrhoea (OR = 3.0 [1.1, 7.8]). Overall, 47% always or often used at least one water treatment. Of the 37% who were very concerned about drinking water, 62% had diarrhoea, 70% always or often used at least one water treatment. An increase in CD4 count was protective only for those with a low risk of diarrhoea associated with medication (OR = 0.6 [0.5, 0.9]). A 30% attributable risk to diarrhoea was estimated for those with high medication risk compared to those with low medication risk. The significant association between concern with drinking water and diarrhoea as well as between concern with drinking water and water treatment suggests awareness that drinking water is a potential transmission pathway for diarrhoeal disease. At the same time we found that a significant portion of diarrhoea was associated with other sources not related to drinking water such as medication usage. PMID- 11895096 TI - A survey of Nipah virus infection among various risk groups in Singapore. AB - Following the Nipah virus (NV) outbreak in March 1999 in Singapore, a serological survey was undertaken to screen individuals potentially exposed to NV. Blood samples were tested for NV IgM, IgG and neutralizing antibodies. Twenty-two (1.5%) of 1469 people tested had antibodies suggesting NV infection. Although 12 of the 22 infected people (54.6%) were symptomatic, the remaining 10 (45.4%) were clinically well and had no past history of compatible pulmonary or neurological disease. Clinical and serological findings suggested three people had been infected with NV before the outbreak was recognized. All those who were infected were male abattoir workers. None of the people who had contact with horses, and no healthcare workers exposed to infected patients and their specimens had detectable antibodies. This study provides evidence that NV causes asymptomatic infection. All of the antibody positive individuals had direct contact with pigs and there was no evidence of human to human transmission. PMID- 11895097 TI - Serological divergence of Dobrava and Saaremaa hantaviruses: evidence for two distinct serotypes. AB - In order to investigate the serological relationship of Dobrava hantavirus (DOBV, originating from Slovenia) and the Dobrava-like Saaremaa virus (SAAV, recently discovered in Estonia) we analysed 37 human serum samples, 24 from Estonia and 13 from the Balkans, by focus reduction neutralization test (FRNT). Most of the Estonian sera (19), including all sera from Saaremaa island (12), reacted with higher FRNT end-point titres to the local SAAV; the majority of them (15 and 11, respectively), with at least fourfold or higher titres to SAAV than to DOBV. In contrast, out of the 13 sera collected in Slovenia, Bosnia-Herzegovina and Greece, only one reacted more strongly with SAAV (with a twofold higher titre), while 10 of these sera reacted more strongly with the local DOBV (9/10 with fourfold or higher titres). These results indicate that DOBV and SAAV define unique hantavirus serotypes. PMID- 11895095 TI - Modelling death rates for carriers of hepatitis B. AB - Hepatitis B carriers who acquired the infection perinatally die from hepatocellular carcinoma (HCC) and cirrhosis at high rates. Published cohort studies are largely limited to males and are too small to estimate the age specific risk of death. We therefore used routinely collected Hong Kong data to estimate the risks. Deaths were partitioned between carriers and non-carriers, then current life table calculations determined life expectancy and probability of dying from HCC or cirrhosis. HCC is the dominant cause of death for male carriers in middle adulthood with a lifetime risk of 27% for HCC compared to 4% for females. Predicted life expectancy is 72 years for male carriers, compared to 79 years for non-carriers. Female carriers have a life expectancy of 81 years and non-carriers 83 years. This model probably applies to all southern Chinese populations and emigrants with similar life history, and other populations that acquired infection early in life. PMID- 11895098 TI - Pharmacogenetics place in modern medical science and practice. AB - Pharmacogenetic evidence-based treatment strategies will have major implications for all aspects of the product pipeline, including drug discovery, high throughput target screening protocols, lead optimization, and drug formulation to produce series of medicines for a particular disease which will meet the efficacy needs of the majority of patients. The initial proof of principle experiments involves whole genome screening for DNA variants and determination of specific patterns of variants associated with adverse events of marketed products [SNP Print(sm)]. Pharmacogenetics has the potential of changing the pipeline model of drug discovery, clinical development, and mass customization marketing. PMID- 11895099 TI - Comparison of metabolic pharmacokinetics of naringin and naringenin in rabbits. AB - Naringin and naringenin are antioxidant constituents of many Citrus fruits. Naringenin is the aglycone and a metabolite of naringin. In order to characterize and compare the metabolic pharmacokinetics of naringenin and naringin, naringenin was administered intravenously and orally to rabbits, and naringin was administered orally. The concentration of naringenin in serum prior to and after enzymatic hydrolysis was determined by HPLC method. The pharmacokinetic parameters were calculated by using WINNONLIN. The results showed that the absolute bioavailability of oral naringenin was only 4%, whereas after taking the conjugated naringenin into account, it increased to 8%. When naringin was administered orally, only little naringenin and predominantly its glucuronides/sulfates were circulating in the plasma. The ratio of AUC of naringenin conjugates to the total naringenin absorbed into the systemic circulation after oral naringenin was much higher when compared to that after i.v. bolus of naringenin, indicating that extensive glucuronidation/sulfation of naringenin occurred during the first pass at gut wall. Oral dosing of naringin resulted in even higher ratio of AUC of naringenin conjugates to the total naringenin than that after oral naringenin. Our results also showed that there were great differences in pharmacokinetics of naringin and naringenin. Oral naringin resulted in latter Tmax, lower Cmax and longer MRT (mean residence time) for both naringenin and its conjugated metabolites than those after oral naringenin. PMID- 11895100 TI - Interaction of digoxin with antihypertensive drugs via MDR1. AB - The multidrug transporter MDR1 (P-glycoprotein)-mediated interaction between digoxin and 29 antihypertensive drugs of various types was examined by using the MDR1 overexpressing LLC-GA5-COL150 cells, which were established by transfecting MDR1 cDNA into porcine kidney epithelial LLC-PK1 cells. These cells construct monolayers with tight junctions, and enable the evaluation of transcellular transport. The MDR1 was highly expressed on the apical membrane (urine side). The basal-to-apical and apical-to-basal transcellular transport of [3H]digoxin in LLC GA5-COL150 cells was time- and temperature-dependent. The basal-to-apical transport of [3H]digoxin was markedly increased, whereas the apical-to-basal transport was decreased in LLC-GA5-COL150 cells, compared with the host LLC-PK1 cells, suggesting that [3H]digoxin was a substrate for MDR1. Most of the Ca2+ channel blockers used here markedly inhibited basal-to-apical transport and increased apical-to-basal transport. Exceptions were diltiazem, nifedipine and nitrendipine, which hardly showed inhibitory effects on transcellular transport of [3H]digoxin. Alpha-blocker doxazosin and beta-blocker carvedilol also inhibited transcellular transport of [3H]digoxin, but none of the angiotensin converting enzyme inhibitors and AT1 angiotensin II receptor antagonists used here were active. These observations will promote understanding of the digoxin drug interactions resulting from their actions on MDR1, and which may aid in avoiding these unexpected effects of digoxin. PMID- 11895101 TI - Influence of sildenafil on central dopamine-mediated behaviour in male rats. AB - Two experiments were performed to evaluate the effects of sildenafil on spontaneous or dopamine agonist-induced behaviour in male rats. Data obtained in experiment 1 show that oral administration of the drug, at 1 mg/kg, significantly increased the occurrence of penile erections, anogenital self-grooming and homosexual mounting in grouped sexually-experienced, but not inexperienced, animals. In experiment 2, pre-treatment with sildenafil (0.5, 1 or 10 mg/kg) dose dependently modified several behavioural signs, centrally evoked by the D2/D3 dopamine agonists, 7-OH-DPAT or B-HT 920 (both at 0.1 mg/kg), in experimentally naive male rats. While sildenafil at 1 mg/kg significantly increased the number of penile erection and stretching-yawning episodes induced by 7-OH-DPAT or B-HT 920, at 10 mg/kg it elicited low stereotyped behaviour, antagonizing stretching yawning and sedation in 7-OH-DPAT treated rats. Discussion centres on the modulatory activity of sildenafil on central dopaminergic pathways and, possibly, on nitric oxide production. PMID- 11895102 TI - Dose-dependent induction of endogenous antioxidants in rat heart by chronic administration of garlic. AB - The inhibitory property of garlic on free radical generation and lipid peroxidation has been reported in a number of in vitro studies. However, the in vivo effects of chronic garlic intake on the antioxidant milieu of heart has not been reported. Therefore, the present study was designed to investigate the effect of chronic garlic homogenate administration on myocardial endogenous antioxidants and lipid peroxidation at five different dosage levels (125, 250, 500, 1000 and 2000 mg/kg; B, C, D, E, F groups respectively). Garlic homogenate was administered orally to Wistar albino rats (150-200 gms) of either sex 6 days/week for 30 days. Myocardial TBARS (Thiobarbituric acid reactive substances) and antioxidants such as SOD (Superoxide Dismutase), catalase, GPx (glutathione peroxidase) and GSH (Reduced Glutathione) were estimated and histopathological changes were observed. Group F was excluded after 55% mortality occurred in 15 days. TBARS levels were significantly lower in groups B, C and D than that of control group (A). Catalase was increased significantly in groups C, D and E, whereas SOD increased significantly in groups B, C and D but decreased in group E. Significant increase in GSH in group E and significant reduction in GPx activity in group B were observed. Histopathological studies showed marked focal myocytolysis in group E. These results showed that chronic garlic intake dose dependently augmented endogenous antioxidants, which might have important direct cytoprotective effects on the heart, especially in the event of oxidant stress induced injury. PMID- 11895104 TI - Effects of phenytoin and/or vitamin K2 (menatetrenone) on bone mineral density in the tibiae of growing rats. AB - In this study, we investigated 1) whether the administration of phenytoin induced bone loss; and 2) whether menatetrenone could prevent bone loss induced by phenytoin. For this purpose, we previously developed a procedure to measure the bone mineral density using a conventional X-ray absorptiometry method. A long termed administration of phenytoin (20 mg/kg per day for 5 weeks) produced bone loss in the tibiae of growing rats. The values of bone mineral density (BMD) were significantly decreased in the tibial diaphysis and metaphysis in the phenytoin treated group. In this period, we measured the serum level of vitamin K-dependent protein, osteocalcin, a marker of bone formation. The serum level of osteocalcin showed a decrease in the phenytoin-treated group compared with the vehicle treated group. Combined administration of menatetrenone (30 mg/kg in diet per day) with phenytoin for 5 weeks prevented the reduction of BMD, and the level of osteocalcin was slightly increased. Thus, it is suggested that long-termed phenytoin exposure may inhibit bone formation concomitantly with insufficient vitamin K, which, at least in part, contributed to bone loss in rats. Finally, these findings implicated the therapeutic usefulness of menatetrenone on a moderate degree of bone abnormality such as drug-induced osteopenia. PMID- 11895103 TI - Molecular characteristics of tissue-bound semicarbazide-sensitive amine oxidase (SSAO) in guinea pig tissues. AB - Various mammalian tissues contain a tissue-bound amine oxidizing enzyme distinct from mitochondrial outer membrane enzyme, monoamine oxidase (MAO, EC 1.4.3.4), termed semicarbazide-sensitive amine oxidase (SSAO, EC 1.4.3.6). An increase in SSAO activity was found in patients suffering from vascular disorders such as diabetes and diabetic complications. It has previously been shown that 2 bromoethylamine (2-BEA) is a potent, and selective suicidal inhibitor of tissue bound SSAO. The aim of this study was to investigate the interaction of this suicidal SSAO inhibitor with the tissue-bound enzyme in guinea pig lung, kidney, stomach, and heart homogenates. The conditions necessary for this inhibitor to titrate the concentrations of this enzyme were also determined. 2-BEA appears to interact with SSAO, as reported previously for this enzyme from different sources, in a manner consistent with an irreversible, "suicide" reaction. Because of this property, 2-BEA could be used to titrate the concentrations of SSAO active centers in these tissues under the appropriate conditions employed. Although some possible non-specific binding of the inhibitor to sites other than the active center of the enzyme, metabolism of this inhibitor and/or presence of enzyme subtypes was hypothesized, the molecular characteristics of SSAO in these tissues (Km, Vmax values, enzyme efficiencies, approximate enzyme concentrations, and molecular turnover numbers) towards the substrate kynuramine (0.1 mM) at pH 7.4 and 37 degrees C have been estimated. PMID- 11895105 TI - Nicotine increases the expression of high affinity nerve growth factor receptors in both in vitro and in vivo. AB - Impairment in nerve growth factor (NGF)-mediated support to basal forebrain cholinergic neurons may represent an initial insult to certain neural cells in Alzheimer's disease (AD). High affinity NGF receptor (TrkA) levels are decreased in AD brains as compared to age-matched control brains. One of the approaches suggested for the treatment of AD exploits the ability of small molecular substances to enhance the expression of endogenous growth factors and/or their receptors. The purpose of this study was to determine whether treatment with nicotine in both in vitro and in vivo settings would increase the neural expression of TrkA receptors. Using a differentiated PC12 neuronal-like system, chronic nicotine treatment increased cell surface TrkA receptor expression. Nicotine's action was blocked by co-treatment with either the non-competitive nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine or with the alpha7 nAChR-selective antagonist methyllycaconitine. Surprisingly, certain low doses of mecamylamine alone also increased TrkA receptor levels. Rats prepared with chronic indwelling intravenous catheters were continuously infused with nicotine to deliver a total dose of 12 mg/kg over 24 hr. This treatment resulted in a significant 44% increase in TrkA receptor expression in the hippocampus. As in the cell experiments, mecamylamine also increased hippocampal TrkA receptor expression. In fact, the ratio of the maximal mecamylamine response to the maximal nicotine response that was measured in vitro, i.e., 0.43 was remarkably similar to that for the in vivo experiment, i.e., 0.47. Since in our previous studies the increase in TrkA expression produced by nicotine was shown to be related to its cytoprotective actions, these results suggest that nicotine's neuroprotective actions might also be mediated through the drug's interaction with central alpha7 nAChRs and subsequent increase in TrkA receptor expression. PMID- 11895106 TI - Type IV collagen prevents amyloid beta-protein fibril formation. AB - The potential of targeting through molecular therapeutics the underlying amyloid beta-protein (A beta) fibrillogenesis causing the initiation and progression of Alzheimer's disease (AD) offers an opportunity to improve the disease. Type IV collagen (collagen IV) is localized in senile plaques in patients with AD. By using thioflavin T fluorescence spectroscopy and electron microscopy, we found that collagen IV inhibited A beta1-40 (A beta40) fibril formation. The critical concentration of collagen IV for this inhibition was 5 microg/mL. Circular dichroism data indicate that collagen IV prevents formation of a beta-structured aggregate of A beta40. These studies demonstrated that collagen IV is apparently a potent inhibitor of A beta fibril formation. PMID- 11895107 TI - Peroxisome proliferator-activated receptor gamma reduces the growth rate of pancreatic cancer cells through the reduction of cyclin D1. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) forms a heterodimeric DNA-binding complex with the retinoid X receptor (RXR) and regulates the transcription of its target genes. Activation of PPARgamma has been shown to induce G1 arrest and to inhibit cell growth of human pancreatic carcinoma cell lines. The purpose of the present study was to examine the effect of ligand activation of PPARgamma and RXR on cell growth and on the expression of G1 cyclins in a pancreatic cancer cell line PANC-1, which expresses PPARgamma at high levels. Troglitazone, a specific ligand for PPARgamma, was found to cause a reduction in the growth rate and induced G1 cell cycle arrest and this effect was additive with that of 9-cis retinoic acid (9-cis RA), a ligand for RXR. Of the G1 cyclins tested, troglitazone specifically reduced the expression of cyclin D1 mRNA and the corresponding protein and this effect was also additive with 9-cis RA. These results suggest that the activation of PPARgamma together with RXR may be useful for the suppression of pancreatic cancer cell growth through the reduction in cyclin D1 levels. PMID- 11895108 TI - The action of dietary phytochemicals quercetin, catechin, resveratrol and naringenin on estrogen-mediated gene expression. AB - Hepatic expression of apolipoprotein (apo) II is in part modulated by estrogen mediated stabilization of its mRNA. This stabilization is due to the estrogen regulated mRNA stabilizing factor (E-RmRNASF) expressed in the liver in response to estrogen (Ratnasabapathy, 1995, Cell. Mol. Biol. Res, 41: 583-594). E-RmRNASF protects the RNA from targeted endonucleolytic degradation. The hepatic expression of E-RmRNASF is modulated by certain estrogenic and antiestrogenic nonsteroidal environmental xenobiotics (Ratnasabapathy et al. 1997, Biochem. Pharmacol., 53: 1425-1434). To determine whether dietary phytochemicals purported to prevent hormone-dependent breast and prostate cancers, and atherosclerosis, acted via the estrogen-cell-signaling pathway, roosters were administered increasing doses up to 1 mmole/kg of resveratrol, quercetin, catechin or naringenin parenterally and tested for hepatic expression of E-RmRNASF. Besides estrogen, the expression of E-RmRNASF in the liver was stimulated by resveratrol and catechin, indicating these agents to be estrogenic. A lack of E-RmRNASF expression was seen with the roosters treated with the vehicle, naringenin or quercetin. To determine whether the agents exerted partial agonistic or antagonistic effects, roosters were administered combinations of estrogen and increasing doses of the above phytochemicals. Resveratrol showed agonistic activity at all concentrations (10-1000 micromol/kg) tested. Catechin showed partial agonistic activity, while quercetin and naringenin appeared to be antagonistic. PMID- 11895109 TI - Development of the lung of the brushtail possum, Trichosurus vulpecula. AB - The developing lung of the brushtail possum, Trichosurus vulpecula, was studied by light microscopy, and transmission electron microscopy was used to study the morphology of the conducting airways in the adult. Bronchi did not extend beyond the hilus of each of the six lobes of the lung, and lobules were supplied by major bronchioles. By 105 days post partum, bronchi and bronchioles were fully formed, coinciding with the emergence of mucosal associated lymphoid tissue (MALT), which preceded alveolar maturation by approximately 20 days. In the adult lung, goblet cells were rarely observed in the mucosal epithelium of bronchi, whereas Clara cells were present in the mucosa of all airways, increasing proportionately as the conducting and respiratory portions narrowed distally. Although the airways of the possum lung have a poorly developed mucociliary blanket, this may be compensated for by the large numbers of Clara cells and adequate supply of MALT. PMID- 11895110 TI - Study of the three-dimensional geometry of the central conducting airways in man using computed tomographic (CT) images. AB - Clinical research on the deposition of inhaled substances (e.g. inhaled medications, airborne contaminants, fumes) in the lungs necessitates anatomical models of the airways. Current conducting airway models lack three-dimensional (3D) reality as little information is available in the literature on the distribution of the airways in space. This is a limitation to the assessment or predictions of the particle deposition in relation to the subject's anatomy. Detailed information on the full topology and morphology of the airways is thus required to model the airway tree realistically. This paper presents the length, diameter, gravity, coronal and sagittal angles that together describe completely the airways in 3D space. The angle at which the airways branch out from their parent (branching angle) and the rotation angle between successive bifurcation planes are also included. These data are from the study of two sets of airways computed tomography (CT) images. One CT scan was performed on a human tracheobronchial tree cast and the other on a healthy male volunteer. The airways in the first nine generations of the cast and in the first six conducting generations of the volunteer were measured using a computer-based algorithm. The data contribute to the knowledge of the lung anatomy. In particular, the spatial structure of the airways is shown to be strongly defined by the central airways with clear angular lobar patterns. Such patterns tend to disappear with a mean gravity, coronal and sagittal angles of 90 degrees in each generation higher than 13-15. The mean branching angle per generation appears independent of the lobe to which the airways belong. Non-planar geometry at bifurcation is observed with the mean (+/- SD) bifurcation plane rotation angle of 79 +/- 410 (n = 229). This angle appears constant over the generations studied. The data are useful for improving the 3D realism of the conducting airway structure modelling as well as for studying aerosol deposition, flow and biological significance of non-planar airway trees using analytical and computational flow dynamics modelling. PMID- 11895111 TI - Differential regional brain growth and rotation of the prenatal human tentorium cerebelli. AB - Folds of dura mater, the falx cerebri and tentorium cerebelli, traverse the vertebrate endocranial cavity and compartmentalize the brain. Previous studies suggest that the tentorial fold has adopted an increasingly important role in supporting the increased load of the cerebrum during human evolution, brought about by encephalization and an adaptation to bipedal posture. Ontogenetic studies of the fetal tentorium suggest that its midline profile rotates inferoposteriorly towards the foramen magnum in response to disproportionate growth of the cerebrum. This study tests the hypothesis that differential growth of the cerebral and cerebellar components of the brain underlies the inferoposterior rotation of the tentorium cerebelli during human fetal development. Brain volumes and tentorial angles were taken from high-resolution magnetic resonance images of 46 human fetuses ranging from 10 to 29 gestational weeks. Apart from the expected increases of both supratentorial and infratentorial brain volumes with age, the results confirm previous studies showing a significant relative enlargement of the supratentorial volume. Correlated with this enlargement was a rotation of the midline section of the tentorium towards the posterior cranial base. These findings support the concept that increases of supratentorial volume relative to infratentorial volume affect an inferoposterior rotation of the human fetal tentorium cerebelli. These results are discussed in the context of the role played by the tentorium cerebelli during human evolution and underline implications for phylogenetic and ontogenetic models of encephalization. PMID- 11895112 TI - Three-dimensional reconstructions of the Achilles tendon insertion in man. AB - The distribution of type II collagen in sagittal sections of the Achilles tendon has been used to reconstruct the three-dimensional (3D) shape and position of three fibrocartilages (sesamoid, periosteal and enthesis) associated with its insertion. The results showed that there is a close correspondence between the shape and position of the sesamoid and periosteal fibrocartilages--probably because of their functional interdependence. The former protects the tendon from compression during dorsiflexion of the foot, and the latter protects the superior tuberosity of the calcaneus. When the zone of calcified enthesis fibrocartilage and the subchondral bone are mapped in 3D, the reconstructions show that there is a complex pattern of interlocking between pieces of calcified fibrocartilage and bone at the insertion site. We suggest that this is of fundamental importance in anchoring the tendon to the bone, because the manner in which a tendon insertion develops makes it unlikely that many collagen fibres pass across the tissue boundary from tendon to bone. When force is transmitted to the bone from a loaded tendon, it is directed towards the plantar fascia by a series of highly orientated trabeculae that are clearly visible in 3D in thick resin sections. PMID- 11895113 TI - Big endothelin-1 but not endothelin-1 is present in the smooth muscle stroma of the prostate gland of the rat. AB - Immunohistochemical techniques were employed to localize the presence of endothelins in the mature rat prostate gland. Immunoreactivity for big endothelin 1 but not endothelin-1 was observed in the fibromuscular stroma of the rat prostate gland. No immunoreactivity was seen in the glandular epithelium. Double staining procedures showed big endothelin-1 immunoreactivity to be co-localized with alpha-actin immunoreactivity. The stroma of the prostate gland also contained nerve fibres coursing through it which are immunopositive for tyrosine hydroxylase. These results suggest that big endothelin-1 but not endothelin-1 is co-localized with alpha-actin in the smooth muscle cells of the rat prostate gland. This implies that endothelin-1 is synthesized on demand from big endothelin-1 in the fibromuscular stroma of the rat prostate. PMID- 11895115 TI - The ramification and connections of retinal fibres in layer 7 of the domestic chick optic tectum: a golgi impregnation, anterograde tracer and GABA-immunogold study. AB - Layer 7 is one of the retinorecipient layers of the avian optic tectum. However, little information is available about the neuronal organization of this layer and its implications for visual function. Golgi impregnation was used to investigate the retinal input to and the neuronal architecture of layer 7 of the chick optic tectum, which forms a narrow band between the two cell-dense layers 6 and 8. Anterograde tracers were also used to investigate the afferent and efferent connections of layer 7, in both the light and the electron microscope, together with GABA immunogold labelling. Three types of radial neuron were defined according to the origin and course of their axons. The perikarya of these neurons were situated in tectal layers 10-11. The principal dendrites of these radial neurons ascended to the tectal surface and gave rise to dendritic side-branches in layer 7. These dendritic side-branches received asymmetric synapses from the terminations of retinal fibre arborisations. Type 2 radial neurons, whose axons arose from the deep pole of the perikaryon or occasionally from a basal dendrite, were shown to project to the nucleus isthmi pars magnocellularis, which has previously been demonstrated to be GABAergic and to project to glomerulus-like complexes in tectal layers 4-5. In these layers, the dendritic branches of layer 13 neurons that project to the nucleus rotundus have previously been shown to receive retinal fibre input. Therefore, the retinal input to layer 7 may be able to modulate the transmission of information to the visual thalamus, by way of a feed-back loop to layers 4-5 of the tectum involving the nucleus isthmi pars magnocellularis. PMID- 11895114 TI - Brief and repeated noise exposure produces different morphological and biochemical effects in noradrenaline and adrenaline cells of adrenal medulla. AB - Exposure to stressful stimuli is known to activate the peripheral sympathetic nervous system and the adrenal gland. In this study, we evaluated the effects of single or repeated bouts of exposure to a readily measurable stressful stimulus (loud noise) on the catecholamine content and ultrastructure of the rat adrenal medulla. In particular, we measured tissue levels of dopamine, noradrenaline, adrenaline and metabolites. In parallel studies, we evaluated the fine ultrastructure of catecholamine cells, including a detailed study of catecholamine granules and a morphometric analysis of adrenaline and noradrenaline medullary cells. Animals were exposed either to a single (6 h) session of loud (100 dBA) noise, or to this noise stimulus repeated every day for 21 consecutive days. There was a marked correlation between biochemical indexes of catecholamine activity and the ultrastructural morphometry of specific catecholamine granules. Exposure to loud noise for 6 h induced a parallel increase in dopamine, noradrenaline, adrenaline and their metabolites, a polarization and an increased numerical density of noradrenaline and adrenaline granules in the cells. After repeated noise exposure, noradrenaline levels were significantly higher than in controls, and adrenaline decreased significantly. In addition, adrenaline cells also exhibited ultrastructural alterations consisting of wide homogeneous cytoplasmic areas and large, pale vesicles. PMID- 11895117 TI - Monolateral hypoplasia of the motor vagal nuclei in a case of sudden infant death syndrome. AB - During the development of motor vagal nuclei (MVN), the neuroblasts of the myeloencephalic basal plate migrate in the dorsolateral direction to form the dorsal motor vagal nucleus (DMVN) and ventrolaterally to form the ventral motor vagal nucleus (VMVN). Those neuroblasts that remain close to the median sulcus will form the hypoglossal nucleus. In support of the congenital origin of the alteration of the MVN in sudden infant death syndrome (SIDS), we report the case of an 8-month-old female child who was found dead in her cot. The neuropathological assessment revealed that the medullary triangle of the 4th ventricle floor was asymmetric, owing to the presence of three prominences to the left side of the median sulcus. The medial prominence corresponded to the hypoglossal nucleus, which showed a marked increase in the number of large neurons; the intermediate prominence corresponded to the DMVN whose large neurons were reduced and were recognizable mainly at the level of the medial fringe; the lateral prominence corresponded to the solitary nucleus. The left solitary tract showed a reduction of the transverse diameter. Also, the left VMVN showed marked reduction in the number of neurons. Inflammatory and astrocytic reactions were absent. We suggest that in SIDS cases the hypocellularity of the MVN and the increased number of neurons of the hypoglossal nucleus are intimately related, indicating a congenital alteration due to incomplete migration of the vagal neuroblasts with abnormality of the autonomic cardio-respiratory control. PMID- 11895116 TI - Mechanisms of changes to the liver pigmentary component during the annual cycle (activity and hibernation) of Rana esculenta L. AB - The present study was performed to elucidate the mechanisms responsible for the changes of melanin content/ distribution we had previously discovered in the liver parenchyma of Rana esculenta during natural hibernation. Melanomacrophagic component response was analysed using morphocytochemical methods. The results demonstrated that during the prehibernation period (October-November) the melanomacrophages reach the highest proliferative activity (BrdU, PCNA labelling) which is accompanied by an evident melanosynthesis (dopa-oxidase activity). In contrast, after hibernation, the decrease of liver pigmentation was the consequence of a partial cell loss by apoptotic mechanisms (TUNEL labelling, pyknosis-karyorhexis) accompanied by a decrease of melanosome content by autophagy and low melanosynthetic activity. On the basis of these findings, there is evidence that liver melanomacrophages represent a metabolically (melanin synthesis/degradation) and cytokinetically (proliferation/ death) active cell population during the annual cycle of the frog. The results are also discussed in relation to the functional synergism between hepatocytes and pigment cells in the adaptation to environmental changes. PMID- 11895118 TI - Versatility and specificity in flavoenzymes: control mechanisms of flavin reactivity. AB - Flavoenzymes are characterized by their remarkable versatility and strict specificity. The former can be grasped when flavoenzymes are treated as a whole, while the latter refers to each flavoenzyme in which the broad versatility of flavin is specifically controlled. The versatility stems from the variety of the redox, ionic, and electronic states that the flavin ring system can adopt. Versatility of flavoenzymes is reflected in their classification, which has generally been based on substrates and reactions catalyzed. A different classification is presented according to the number of electrons transferred in the reductive and oxidative half reactions. Specificity of each flavoenzyme is understood in terms of the regulatory mechanism of the broad reactive potentiality of flavin. The elements of this regulatory mechanism include hydrogen-bonding network, electrostatic effect, charge-transfer interaction, positioning between a substrate/ligand and flavin, and modulation of resonance hybridization, each of which is explained with relevant examples provided mainly by studies from the author's group. PMID- 11895119 TI - Chemistry and chemical biology of taxane anticancer agents. AB - Taxol (paclitaxel) and Taxotere (docetaxel) are currently considered to be among the most important anticancer drugs in cancer chemotherapy. The anticancer activity of these drugs is ascribed to their unique mechanism of action, i.e., causing mitotic arrest in cancer cells, leading to apoptosis through inhibition of the depolymerization of microtubules. Although both paclitaxel and docetaxel possess potent antitumor activity, treatment with these drugs often results in a number of undesirable side effects, as well as multidrug resistance (MDR). Therefore, it has become essential to develop new anticancer agents with superior pharmacological properties, improved activity against various classes of tumors, and fewer side effects. This paper describes an account of our research on the chemistry of paclitaxel and taxoid anticancer agents at the biomedical interface, including: 1. The structure-activity relationship (SAR) study of taxoids leading to the development of the "second-generation" taxoids, which possess exceptional activity against drug-resistant cancer cells expressing the MDR phenotype. 2. Development of fluorinated taxoids to study the bioactive conformation of paclitaxel and photoaffinity labeling taxoids for mapping of the drug-binding domain on both microtubules and P-glycoprotein. 3. The synthesis of novel macrocyclic taxoids for the investigation into the common pharmacophore for microtubule stabilizing anticancer agents. PMID- 11895120 TI - Reflections on the discovery of nature's pathways to vitamin B12. AB - The chronology of the discoveries along the pathway of vitamin B12 biosynthesis is reviewed from a personal perspective, including discussion of the most recent finding that two pathways to B12 exist--one aerobic and one anaerobic--which differ mainly in the ring contraction mechanisms which convert porphyrin to corrin. PMID- 11895121 TI - The chemistry and biological function of natural marine toxins. AB - Studies on ciguatera fish poisoning led to clarification of the absolute stereochemistry of ciguatoxin, gambierol, gambieric acids, and maitotoxin. Anisotropic NMR reagents and fluorometric chiral HPLC reagents were effectively used together with synthesis of partial structures. Structures of 16 ciguatoxin congeners were successfully elucidated by FAB/MS/MS using samples of 5 microg or less. Stereochemical assignments were also achieved on dinophysistoxin-1, pectenotoxins, yessotoxins, polycavernoside-A, azaspiracid, and prymnesins. The toxins possessed poly-cyclic-ether structures and originated from unicellular algae. Biological functions are briefly described. PMID- 11895122 TI - 1- , 2-, and 3-dimensional polyphenylenes--from molecular wires to functionalised nanoparticles. AB - Polyphenylenes, ranging from one-dimensional wire-like conjugated polymers to two dimensional disc-shaped polyaromatic hydrocarbons and three-dimensional sphere like dendrimers, have been prepared using methods that allow synthetic control of their molecular and supramolecular order in order to optimise their physical, especially optical and electrical, properties. 1D-conjugated polymers can be used as emitting materials in LEDs, with their colours tuned so as to give emission across the whole visible spectrum. Their supramolecular order can be manipulated by attachment of bulky sidechains to suppress aggregation, and by formation of rod-coil block copolymers. 2D polycyclic aromatic hydrocarbons form stable columnar mesophases with high charge carrier mobilities. Their size, shape, and substitution patterns can be altered so as to maximise their intra- and intercolumnar order. 3D polyphenylene dendrimers can be prepared in ways that enable control of their shape, and their surfaces can be selectively functionalised in ways that permit them to act as functionalised nanoparticles. PMID- 11895123 TI - Ultrafast dynamics of myoglobin probed by time-resolved resonance Raman spectroscopy. AB - Recent experimental work carried out in this laboratory on the ultrafast dynamics of myoglobin (Mb) is summarized with a stress on structural and vibrational energy relaxation. Studies on the structural relaxation of Mb following CO photolysis revealed that the structural change of heme itself, caused by CO photodissociation, is completed within the instrumental response time of the time resolved resonance Raman apparatus used (approximately 2 ps). In contrast, changes in the intensity and frequency of the iron-histidine (Fe-His) stretching mode upon dissociation of the trans ligand were found to occur in the picosecond regime. The Fe-His band is absent for the CO-bound form, and its appearance upon photodissociation was not instantaneous, in contrast with that observed in the vibrational modes of heme, suggesting appreciable time evolution of the Fe displacement from the heme plane. The band position of the Fe-His stretching mode changed with a time constant of about 100 ps, indicating that tertiary structural changes of the protein occurred in a 100-ps range. Temporal changes of the anti Stokes Raman intensity of the v4 and v7 bands demonstrated immediate generation of vibrationally excited heme upon the photodissociation and decay of the excited populations, whose time constants were 1.1 +/- 0.6 and 1.9 +/- 0.6 ps, respectively. In addition, the development of the time-resolved resonance Raman apparatus and prospects in this research field are described. PMID- 11895124 TI - Brain oxidative stress--analytical chemistry and thermodynamics of glutathione and NADPH. AB - Oxidative stress occurs in the brain due to stroke, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, trauma, aging and other conditions. Analysis of the effects of oxidative stress can involve quantitation of brain GSH, GSSG, NADPH and NADP. Reliable and rapid assays have been developed for these compounds and will be presented in detail. The assays have been used to analyze the effects of brain oxidative stress. Thermodynamic calculations can be performed to find the observed electrochemical potentials of the GSSG/GSH and the NADP/NADPH couples during oxidative stress. The biochemical consequences of these thermodynamic changes in the cell will be discussed as well as the defense mechanisms available to the cell to recover from oxidative stress. PMID- 11895125 TI - DNA damage and repair in the brain after cerebral ischemia. AB - In experimental models of brain injury of the ischemia-reperfusion type, there is a period of time in which the formation of oxidative damage exceeds its repair. Simultaneously, the expression of immediate early genes is induced to activate the expression of late effector genes. Drugs that reduce the need to repair during this transient period of time also attenuate neuronal death after brain injury. An example discussed in this review is the activator protein-1 (AP-1), the product of the c-fos gene and other immediate early genes. What is the effect of a delayed expression of these genes in relationship to the process of cell death? This short period presents a window of opportunity to study the effects of oxidative damage on gene expression in the brain and specific deficiencies in gene repair that have been associated with particular neurological disorders. PMID- 11895126 TI - Oxytosis: A novel form of programmed cell death. AB - Extensive nerve cell death occurs during the development of the central nervous system as well as in episodes of trauma and in neurodegenerative disease. The mechanistic details of how these cells die are poorly understood. Here we describe a unique oxidative stress-induced programmed cell death pathway called oxytosis, and outline pharmacological approaches which interfere with its execution. Oxidative glutamate toxicity, in which exogenous glutamate inhibits cystine uptake through the cystine/glutamate antiporter leading to a depletion of glutathione, is used as an example of oxytosis. It is shown that there is a sequential requirement for de novo macromolecular synthesis, lipoxygenase activation, reactive oxygen species production, and the opening of cGMP-gated channels which allow the influx of extracellular calcium. The translation initiation factor elF2alpha plays a central role in this pathway by regulating the levels of glutathione. Finally, examples are given in which the reduction in glutathione, the production of reactive oxygen species, and calcium influx can be experimentally manipulated to prevent cell death. Data are reviewed which suggest that oxytosis may be involved in nerve cell death associated with nervous system trauma and disease. PMID- 11895127 TI - Production of reactive oxygen species from aggregating proteins implicated in Alzheimer's disease, Parkinson's disease and other neurodegenerative diseases. AB - The deposition of abnormal protein fibrils is a prominent pathological feature of many different 'protein conformational' diseases, including some important neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), motor neurone disease and the 'prion' dementias. Some of the fibril-forming proteins or peptides associated with these diseases have been shown to be toxic to cells in culture. A clear understanding of the molecular mechanisms responsible for this toxicity should shed light on the probable link between protein deposition and cell loss in these diseases. In the case of the beta amyloid (Abeta), which accumulates in the brain in AD, there is good evidence that the toxic mechanism involves the production of reactive oxygen species (ROS). By means of an electron spin resonance (ESR) spin-trapping method, we have shown recently that solutions of Abeta liberate readily detectable amounts of hydroxyl radicals upon incubation in vitro followed by the addition of small amounts of Fe(II). We have also obtained similar results with alpha-synuclein, which accumulates in Lewy bodies in PD. Our data suggest that hydrogen peroxide accumulates during Abeta or alpha-synuclein incubation and that this is subsequently converted to hydroxyl radicals, on addition of Fe (II), by Fenton's reaction. Consequently, we now support the idea that one of the fundamental molecular mechanisms underlying the pathogenesis of cell death in AD, PD, and possibly some other protein conformational diseases, could be the direct production of ROS during formation of the abnormal protein aggregates. This hypothesis suggests a novel approach to the therapy of this group of diseases. PMID- 11895128 TI - Dopamine thioethers in neurodegeneration. AB - Dopamine oxidation is proposed to be a significant contributor to mesostriatal dopaminergic neurodegeneration, although the mechanisms are not fully resolved. Recent results from in vitro and in vivo models have suggested that some products from mercapturic acid pathway (MAP) metabolism of oxidized dopamine (DA) may contribute to dopaminergic neurodegeneration, and that at least one product of this pathway, 5-S-cysteinyldopamine (Cys-DA), is elevated in patients with advanced Parkinson's disease (PD). Here we review recent findings on MAP enzymes and their products in rodent brain and in diseased regions of brain from patients with mesostriatal dopaminergic neurodegeneration. We also review the current data and our recent findings on the neurobiological activity of MAP metabolites of oxidized DA. We conclude that human striatum has limited enzymatic capacity for mercapturate formation, that levels of MAP products of oxidized DA are significantly elevated in PD patients with advanced dopaminergic neurodegeneration but not in patients with less severe degeneration, and that Cys DA interferes with trafficking of DA in vitro and in vivo. These results indicate that Cys-DA may interfere with DAtrafficking in patients with advanced dopaminergic neurodegeneration. PMID- 11895129 TI - Toxic metals and oxidative stress part I: mechanisms involved in metal-induced oxidative damage. AB - Toxic metals (lead, cadmium, mercury and arsenic) are widely found in our environment. Humans are exposed to these metals from numerous sources, including contaminated air, water, soil and food. Recent studies indicate that transition metals act as catalysts in the oxidative reactions of biological macromolecules therefore the toxicities associated with these metals might be due to oxidative tissue damage. Redox-active metals, such as iron, copper and chromium, undergo redox cycling whereas redox-inactive metals, such as lead, cadmium, mercury and others deplete cells' major antioxidants, particularly thiol-containing antioxidants and enzymes. Either redox-active or redox-inactive metals may cause an increase in production of reactive oxygen species (ROS) such as hydroxyl radical (HO.), superoxide radical (O2.-) or hydrogen peroxide (H2O2). Enhanced generation of ROS can overwhelm cells' intrinsic antioxidant defenses, and result in a condition known as "oxidative stress". Cells under oxidative stress display various dysfunctions due to lesions caused by ROS to lipids, proteins and DNA. Consequently, it is suggested that metal-induced oxidative stress in cells can be partially responsible for the toxic effects of heavy metals. Several studies are underway to determine the effect of antioxidant supplementation following heavy metal exposure. Data suggest that antioxidants may play an important role in abating some hazards of heavy metals. In order to prove the importance of using antioxidants in heavy metal poisoning, pertinent biochemical mechanisms for metal induced oxidative stress should be reviewed. PMID- 11895131 TI - Oxidative stress in neurodegeneration: mechanisms and therapeutic perspectives. AB - Neural tissue is especially sensitive to oxidative stress, which is considered a prominent factor in both acute and chronic neurodegenerative diseases and traumatic brain insults. On this basis, therapeutical strategies centered on antioxidants and on drugs able to scavenge excess free radicals and to re establish the redox equilibrium, have been proposed for treatment of several brain pathologies. The present paper shortly summarizes the main sources of free radical production in the brain and reviews some of the recent data on mechanisms of cellular transduction through which free radicals are believed to damage cells and, eventually, to bring them to death. Some of the most promising therapeutical perspectives for treatment of oxidative stress in neurodegeneration, are then considered. Their choice is the result of a selection, that is unavoidably due to the enormous amount of the literature data, based on personal evaluation as well as on the personal experimental experience of the author. Four main categories of possible therapeuticals are considered: inhibitors of antioxidant enzymes, endogenous antioxidants and their precursors, vitamins and related compounds, other natural antioxidants from fruits and vegetables. Some theoretical and practical issues relevant to the adoption of antioxidant therapies for neurodegeneration are highlited, with particular reference to the fact that a basal production of free radicals must be maintained in the brain due to the host of essential cellular functions subserved by them. In this connection, it seems advisable that future antioxidant strategies for neurodegeneration are based on mixtures of agents able to modulate multiple mechanisms of free radical production and scavenging, without dangerously hampering essential physiological defense based on free radical cellular signaling. PMID- 11895132 TI - Role of flavonoids in oxidative stress. AB - Flavonoids are a group of naturally occuring compounds which are widely distributed in nature. Epidemiological evidence suggests an inverse relationship between dietary intake of flavonoids and cardiovascular risk. The biological activities of flavonoids are related to their antioxidative effects. But a number of studies have found both anti and prooxidant effects for many of these compounds. This review article presents the synthetic pathways of flavonoids and discusses the structure-activity relationships between, xanthine oxidase inhibitive activities and their chemical structures, between the antioxidant and prooxidant activities and the chemical structure. Then we will show the antioxidant properties of new flavonoids in a few models. In these compounds one or two di-tert-butylhydroxyphenyl (DBHP) groups replace the catechol moiety at the position 2 of the benzopyrane heterocycle. New structures are compared with quercetin and BHT in an LDL-oxidation system, in protecting cultured bovine aortic endothelial cells against mO-LDL cytotoxicity and on myocardial functional recovery during reperfusion after 30 min global ischemia in isolated rat hearts. PMID- 11895130 TI - The chemistry of transition metals in relation to their potential role in neurodegenerative processes. AB - Cells rely on several transition metals to regulate a wide range of metabolic and signaling functions. The diversity and efficiency of their physiological functions are derived from atomic properties that are specific to transition metals, most notably an incomplete inner valence subshell. These properties impart upon these elements the ability to fluctuate among a variety of positively charged ionic forms, and a chemical flexibility that allows them to impose conformational changes upon the proteins to which they bind. By this means, transition metals can serve as the catalytic centers of enzymes for redox reactions including molecular oxygen and endogenous peroxides. This review addresses the consequences of the aberrant translocation of the redox-capable essential transition elements, iron, copper, and manganese, upon the brain with an emphasis on uncontrolled and deleterious oxidative events. The potential of metal-protein interactions in facilitating such events, and their association with the physiologically redox-inert metals zinc and aluminum, are related to their postulated contribution to the pathology of neurodegeneration. PMID- 11895134 TI - Regional anesthesia and analgesia: their role in postoperative outcome. AB - Increasing insight into the mechanisms of perioperative physiologic responses and the resultant effects on patient outcome suggests that some responses may be detrimental to long-term recovery. Thus, initial belief in the adaptive "wisdom" of the body has been supplanted by the concept that a "stress-free" perioperative period may minimize detrimental physiologic responses and resultant morbidity. The perioperative use of neuraxial- or regional anesthesia and analgesia have profound inhibitory effects on the body's response to surgery compared to the same operation performed during general anesthesia alone. Increasing evidence has emerged suggesting that such afferent nociceptive blockade may improve a variety of postoperative morbidity parameters and improve surgical outcome. We review the clinical evidence from studies examining the effects of regional anesthesia and analgesia on postoperative morbidity in specific physiologic systems. PMID- 11895133 TI - From cocaine to ropivacaine: the history of local anesthetic drugs. AB - In 1850, about three centuries after the conquest of Peru by Pizzaro, the Austrian von Scherzer brought a sufficient quantum of coca leaves to Europe to permit the isolation of cocaine. As suggested by his friend Sigmund Freud, descriptions of the properties of the coca prompted the Austrian Koller to perform in 1884 the first clinical operation under local anesthesia, by administration of cocaine on the eye. The use of cocaine for local and regional anesthesia rapidly spread throughout Europe and America. The toxic effects of cocaine were soon identified resulting in many deaths among both patients and addicted medical staff. Local anesthesia was in a profound crisis until the development of modern organic chemistry which led to the synthesis of pure cocaine in 1891. New amino ester local anesthetics were synthesized between 1891 and 1930, such as tropocaine, eucaine, holocaine, orthoform, benzocaine, and tetracaine. In addition, amino amide local anesthetics were prepared between 1898 and 1972 including nirvaquine, procaine, chloroprocaine, cinchocaine, lidocaine, mepivacaine, prilocaine, efocaine, bupivacaine, etidocaine, and articaine. All of these drugs were ostensibly less toxic than cocaine, but they had differing amounts of central nervous system (CNS) and cardiovascular (CV) toxicity. Bupivacaine is of special interest because of its long duration of action and history of clinical application. Synthesized in 1957, the introduction of bupivacaine on the market in 1965 paralleled the progressive and cumulative reports of CNS and CV toxicity, leading to the restriction of its use and the identification of a special therapy-resistant CV toxicity. Numerous experimental studies were conducted to identify the fine cellular mechanism of this toxicity, which refines our understanding of the action of local anesthetics. The identification of optically active isomers of the mepivacaine family led to the selection of ropivacaine, a pure S-(-) enantiomer, whose toxicology was selectively and extensively studied before its introduction on the market in 1996. During the rapid and extensive use of ropivacaine in the clinic, unwanted side-effects have been found to be very limited. PMID- 11895135 TI - Alpha2-adrenergic receptor agonists as analgesics. AB - Alpha2-adrenergic receptor agonists are analgesic agents, and the alpha2 adrenergic agonist clonidine has been used in clinical studies for regional analgesia after intrathecal administration. We review here recent developments concerning the structure activity relationships of a new class of potent alpha2 adrenergic agonists and their use as analgesic agents. The effect of structure upon cardiovascular side-effects is also monitored, such as the prolongation of the QT portion of the cardiac action potential. PMID- 11895136 TI - Nerve injury associated with regional anesthesia. AB - Neural damage is a possible consequence of general anesthesia, central nervous system blockade, and regional anesthesia. Dainage may be caused by ischaemic and mechanical or chemical factors, which may occur either alone or in combination. Neural damage may be secondary to prolonged and severe arterial hypotension compromising blood supply to the cord, a spinal haematoma whose main etiological factor is a coagulation abnormality, an intraneural injection, and peripheral neuropathy related to perioperative positioning. Mechanical trauma by the needle bevel is an important factor contributing to neuropathy. Neurological complications may also result from a direct neurotoxic effect of local anesthetic agents which is concentration and dose-dependent. A better understanding of these mechanisms will provide a reliable basis for the development of improved pharmaceutical therapy. PMID- 11895137 TI - The enantiomers: revolution or evolution. AB - The use of single stereoisomers are gaining popularity in the world of anesthesiology. The reduced costs of production have made these compounds available for clinical application. The majority of drugs used in anesthesiology such as ketamine, isoflurane, etomidate, atracurium, bupivacaine and ropivacaine have an asymmetric carbon, and are still used primarily as racemic mixtures (1:1 mixture of R and S enantiomers). Among local anesthetics, the S enantiomers often have favorable biological properties. This insight has led to the development of ropivacaine. Ropivacaine is the first local anesthetic marketed as pure S-(-) enantiomer. Its pharmacodynamic and pharmacokinetic profile is similar to that of bupivacaine, but in vitro and in vivo studies have shown that ropivacaine is less cardiotoxic. Clinical data suggests that ropivacaine has a greater margin of safety than bupivacaine, which is necessary for further expanding the application of regional anesthesia. The time has come for the use of single enantiomers in regional and general anesthesia PMID- 11895138 TI - The cardiotoxicity of local anesthetics: the place of ropivacaine. AB - Central and regional block procedures have a well-defined role as safe and effective methods in modern anesthesia and analgesia with long-acting local anesthetics. Recent studies have shown that the incidence of intoxication by these drugs is a rare but catastrophic event. As classic neuronal sodium channel inhibitors, local anesthetics block peripheral fast voltage-gated sodium channels on neuronal axons, and these drugs have a particularly high level of activity in the CNS and the cardiovascular system. CNS-toxicity follows a two-stage process, whereby at lower concentrations inhibitory neurons are blocked first resulting in generalized convulsions, and at higher concentrations a global CNS depression can be seen. Although seizures are an impressive clinical syndrome, they can often be treated safely without permanent damage. More important is the cardiotoxicity of these drugs, which can be divided into indirect cerebrally mediated and a direct myocardial component. Like CNS-toxicity in general, indirect cardiotoxicity demonstrates an initial stimulating effect, followed by a depressive component at higher concentrations. Direct myocardial actions are comprised of negative chronotropic, dromotropic and inotropic effects. For dromotropy, stereoselectivity was found. The S-(-)-isomers of the longacting local anesthetics were less delayed compared to racemic mixtures and the R-(+) enantiomers. For inotropy, no stereospecific depression of this parameter was noted between isomers of ropivacaine or bupivacaine, but bupivacaine produced a significantly greater depression of LV pressure than ropivacaine, mepivacaine, or lidocaine. Pharmacokinetic differences in lipophilicity of local anesthetics correlate well with the depression mitochondrial ATP-synthesis in fast metabolizing cells. Intracellular ATP-level may be involved in contractility and resuscitation of cardiomyocytes, as be proven by in-vitro and in-vivo data. Therefore the use of pure optical S-(-)-isomers of local anesthetics may help to reduce these rare but catastrophic events. Presently, ropivacaine appears to be the safest long-acting local anesthetic. PMID- 11895139 TI - Clinical application of ropivacaine in obstetrics. AB - Ropivacaine is a new long-acting local anesthetic which is a pure (S)-(-) enantiomer, with an efficacy profile similar to that of bupivacaine. Compared in equal doses, ropivacaine shows more separation between sensory and motor blockade than bupivacaine. Moreover, ropivacaine has a lower systemic toxicity than bupivacaine. In obstetrics, ropivacaine and bupivacaine have been compared for Cesarean section and for epidural pain relief during labor and delivery. For Cesarean section, both drugs provide similar analgesia when given in equal doses, but motor block is less pronounced with ropivacaine. Neonatal outcome as determined by Apgar scores and Neurological Adaptive Capacity Scores (NACS) is also similar. For epidural pain relief during labor and delivery, both drugs are equally effective, either when given alone or in combination with opioids; a meta analysis of six studies showed that compared to bupivacaine, the use of ropivacaine is associated with significantly less motor block and instrumental deliveries. PMID- 11895140 TI - Clinical application of ropivacaine for the upper extremity. AB - Ropivacaine, the S-(-)-enantiomer of N-(2,6-dimethylphenyl)-1-propyl-2 piperidinecarboxamide is a new long-acting local anesthetic. This review demonstrates that it is effective in brachial plexus anesthesia. It is at least as efficient as bupivacaine in terms of quality, duration of analgesia, anesthesia, and motor block. It could have some advantages over bupivacaine in terms of onset time of sensory and motor block, but this remains controversial. In single-shot brachial plexus block, it is equipotent to bupivacaine and has a similar pharmacokinetic profile. Its minimal effective concentration is 0.5%, and the benefit of increasing its concentration to 0.75 or 1% remains debatable. Its use during continuous brachial plexus block has been much less studied, and conflicting results involving efficacy during continuous interscalene block and inefficacy during continuous axillary block have been obtained. Further investigations are required to assess its efficacy during such block. Because of lower CNS and cardiac toxicity, ropivacaine is safer than bupivacaine. It would be thus the preferred local anesthetic for brachial plexus blockade when long lasting anesthesia and analgesia is required. PMID- 11895141 TI - Clinical application of ropivacaine for the lower extremity. AB - Ropivacaine is a new amide local anaesthetic, which is the first commercially available in its category as a pure S-(-) enantiomer. In most recent studies, ropivacaine exhibited a very close pharmacodynamic profile to equipotent doses of bupivacaine. Concentrations of 0.5%, 0.75% and 1% (5, 7.5 and 10 mg/mL, respectively) ropivacaine are used for intraoperative anaesthesia, while the concentration of 0.2% (2 mg/mL) is preferred for postoperative analgesia, either alone or in combination with opioids and/or clonidine. Ropivacaine is responsible for excellent postoperative analgesia following epidural and peripheral perineural injections, using single-shot injections and continuous infusions. Differential sensory/motor block is only apparent at low concentrations (0.2% and less). A significant amount of recent literature focuses on its use for peripheral blocks of the lower limbs, i.e. sciatic and femoral nerve blocks. The primary benefit of ropivacaine is its lower toxicity, mainly lower cardiotoxicity, following accidental intravascular injection. This higher therapeutic index leads to an improved safety profile as compared with potent local anaesthetics such as racemic bupivacaine. For that reason, ropivacaine is a good choice for both intraoperative and postoperative regional anaesthesia and analgesia. PMID- 11895142 TI - Mushroom bodies and post-mating behaviors of Drosophila melanogaster females. AB - After mating, Drosophila melanogaster females lay substantially more eggs and mate rarely. Central to these changes is Sex peptide (SP), a male peptide transferred into the female during copulation. Injected into virgins, SP induces the same post mating response as observed after mating. In this study we investigated the role of the mushroom body (MB) in the SP response system. The SP response of females with either chemically ablated or mutant MBs was analyzed. After injection of SP, females with chemically ablated MBs reduce their receptivity and increase their ovulation and oviposition to the level of females with intact MBs. Virgin females with ablated MBs, however, show a constitutively elevated oviposition rate. Hence in untreated females, MBs are not implicated in the SP-induced reduction of receptivity and increase of ovulation. However, they depress the oviposition rate of virgins. Thus, SP has two functions for oviposition: it de-represses the MB-dependent block on the egg laying activity of virgins and additionally stimulates oviposition. SP-injected mushroom body miniature (mbm) females lay fewer eggs, ovulate less frequently, and mate more often than wild-type females. A model of the putative role of MBs and the gene product of mbm in SP-induced oviposition is presented. PMID- 11895143 TI - Generation of a semi-dominant mutation with temperature sensitive effects on both locomotion and phototransduction in Drosophila melanogaster. AB - A novel Drosophila mutant named Tripped-and-fell (Taf) was isolated in a F1 screen for dominant temperature sensitive paralytics. Recombination mapping using multiply marked chromosomes and P elements have pinpointed the locus of Taf to polytene band 93 on the right arm of the third chromosome (3R). When exposed to restrictive temperatures, both Taf heterozygotes and homozygotes paralyzed; however, homozygotes paralyzed at lower temperatures and took longer to recover than heterozygotes. There are also positive correlations between recovery time from paralysis and both duration and temperature of exposure. Electroretinograms (ERGs) revealed that both homozygotes and heterozygotes have a grossly normal light response at 22 degrees C, but at 37 degrees C, the ERGs from both homozygotes and heterozygotes are unable to maintain a normal sustained depolarization and have a reduced off-transient potential. The severity of the ERG repolarization phenotype is greater in homozygotes than in heterozygotes. PMID- 11895144 TI - Extra ocular photic entrainment in Drosophila melanogaster. AB - The ability to anticipate the daily changes occurring in the photic environment, by adjusting their physiology and behavior accordingly, should provide living organisms with a selective advantage. Organisms could thus sample the relevant entraining stimuli at early dawn and/or late dusk. At these times of the day the principal spectral components of sunlight reaching the Earth's surface consist of relatively low levels of irradiance in the blue and red parts of the visible spectrum. Biological circadian systems could entrain to twilight conditions by sampling either one or both of these components. Previous studies, based on the characterization of phase response curves, following brief exposures to relatively high intensity light of varying wavelength, demonstrated the particular sensitivity of the Drosophila circadian system to blue light. In this study, we addressed the capacity of the circadian system of Drosophila to respond to long periods of exposure to red light. Thus flies were initially exposed to 12 h : 12 h LD cycles with full spectrum white light. Following a 1-4 h phase shift, the flies were exposed to LD cycles with red light. Our results suggest that, under these experimental conditions, red light of wavelength between 650-700 nm, can function as an entraining stimulus. Furthermore, analysis of the circadian locomotor activity profiles in visually impaired flies, suggests that in wild type flies locomotor activity is triggered by the circadian clock at key times during the day. Once triggered, the whole cycle (i.e. onset, peak and offset) of locomotor activity occurring both at dawn and dusk can proceed autonomously. However, the occurrence of a lights-off signal (typically at dusk) before the autonomous cessation of locomotor activity, leads to a light-driven termination of such activity. In addition, so1 (eyeless) mutant flies show the presence of a single evening locomotor activity peak during the whole circadian day, suggesting that in wild type flies the morning and evening activity peaks may be under separate control. PMID- 11895145 TI - Immunonutrition. PMID- 11895146 TI - Impact of fish oil enriched total parenteral nutrition on DNA synthesis, cytokine release and receptor expression by lymphocytes in the postoperative period. AB - A prospective randomized study on sixty patients was conducted to investigate the effects of a fish oil containing total parenteral nutrition (TPN) regimen in the postoperative period on lymphocyte subset distribution, proliferation, cytokine production and interleukin-2 receptor (IL-2R) expression. Patients who underwent large bowel surgery were divided into three groups. Nineteen patients received TPN with fish oil (0.2 g/kg body weight per day) plus soybean oil (1.0 g/kg per day), twenty patients received soybean oil (1.2 g/kg per day), and twenty-one patients who were on a fat-free regimen served as the control group. Natural killer (NK) cells, total, B-, T-, T4-, T8-lymphocytes, proliferation of lymphocytes, in vitro production of IL-2, IFN-gamma, TNF-alpha, and IL-2R expression were measured. Fish oil administration did not affect subset distribution and proliferation of lymphocytes. Production of interleukin-2 (IL 2), interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) was augmented, and IL-2R expression less enhanced compared with the controls. It is concluded that administration of 0.2 g/kg per day fish oil after a moderate surgical stress is not immunosuppressive, but enhances the production of IFN gamma, TNF-alpha and possibly IL-2. PMID- 11895147 TI - Nutrition and HIV infection. AB - Infection by the human immunodeficiency virus (HIV) is characterized by progressive destruction of the immune system, which leads to recurrent opportunistic infections and malignancies, progressive debilitation and death. Malnutrition is one major complication of HIV infection and is recognized as a significant prognostic factor in advanced disease. Malnutrition is multifactorial and poorly treated during the course of HIV. Even if a standardized approach to the management of active weight loss has not been well established, early nutritional intervention is important in HIV infected patients to maximize gain of lean body mass. From early in the era of highly active antiretroviral therapy (HAART), an initial decreased incidence of malnutrition was noted only in western countries while a variety of changes in the distribution of body fat and associated metabolic abnormalities have been recognized under the banner of lipodystrophy. PMID- 11895148 TI - Immune-modulatory actions of arginine in the critically ill. AB - Current trials of immune-enhancing diets suggest several beneficial clinical effects. These products are associated with a reduction in infectious risk, ventilator days, ICU and hospital stay. However, methodological weaknesses limit the inferences we can make from these studies. Furthermore, improvements in outcomes were largely seen in surgical patients and in patients who tolerated critical amounts of formula. We propose that the beneficial findings cannot easily be extrapolated to other patient populations since there is suggestion from clinical trials that the sickest patients, especially those with severest appearances of sepsis, shock and organ failure may not benefit or may even be harmed. In these conditions we hypothesize that systemic inflammation might be undesirably intensified by immune-enhancing nutrients like arginine in critically ill patients. In this paper, we review the purported effects of arginine on the immune system and organ function to understand the scientific rationale for its inclusion into enteral feeding products. We conclude that patients with the most severe appearances of the systemic inflammatory response syndrome should not receive immune-enhancing substrates which may aggravate systemic inflammation and worsen clinical outcomes. PMID- 11895149 TI - Immunonutrition in patients after multiple trauma. AB - Severe trauma threatens the life of the victim, both directly and indirectly via immunological dysregulation during the subsequent clinical course. Inflammatory or infectious episodes may complicate the clinical course and ultimately result in sepsis and multiple organ failure, which have mortality rates of up to 80%. Immunomodulatory intervention aims to ameliorate the early hyperinflammatory phase (systemic inflammatory response syndrome, SIRS) to avoid the development of sepsis. One of the immunomodulation strategies is enteral feeding supplemented with specific nutrients, such as glutamine, n-3-polyunsaturated fatty acids, and nucleotides ('immunonutrition'), because changes in the GALT (gut-associated lymphoid tissue) immune response may contribute to intestinal dysfunction and increase susceptibility to post injury gut-derived sepsis. In a prospective, randomized, double-blind, controlled study in twenty-nine patients suffering severe trauma we were able to show that immunonutrition (arginine, n-3-fatty acids, and nucleotides) significantly reduces the number of SIRS days per patient, and also lowers the multiple organ failure (MOF) score on day 3 and days 8-11 (P<0.05). Other studies have reported a reduction in septic complications and MOF rates, shortened hospital stay, and reduction in the use of antibiotics in patients randomized to the immune-enhancing diet. This improved clinical outcome was reflected in a reduction in hospital costs. In the recovery period after trauma (1-72 h after injury) a limitation of the inflammatory response of immunocompetent cells must be achieved as quickly as possible (<72 h). The only strategy available to clinicians caring for trauma patients is immunonutrition, and this should be strongly considered as a rational approach improving immune function and reducing septic complications in critically ill or injured patients. PMID- 11895150 TI - Nucleotides as semiessential nutritional components. AB - Dietary nucleotides are required nutrients for some tissues under certain circumstances. A lack of dietary nucleotides negatively influences protein synthesis in both the liver and the small intestine of rats. Ribosome degradation has been observed as being among the mechanisms responsible for this effect. Dietary nucleotides can also modulate gene expression by interaction with specific transcription factors, in both the liver and the small intestine. PMID- 11895151 TI - Glutamine depletion impairs cellular stress response in human leucocytes. AB - During sepsis and major trauma the blood glutamine (Gln) level is reduced. The administration of Gln can improve the outcome of these patients. However, the mechanism of this beneficial effect of Gln is poorly understood. In the course of critical illness leucocytes are confronted with cytotoxic inflammatory mediators. To protect themselves against these factors, cells express heat shock proteins (HSP). Previous studies have shown that the expression of the major inducible HSP (HSP70) is improved by high Gln concentrations above 4 mM. In this study we investigated whether Gln depletion, such as observed during critical illness, has an effect on HSP70 expression. Human lymphocytes exposed for 2 h to 42 degrees C showed a 3-fold increase in HSP70 expression (P<0.01). A preceding Gln starvation period over 3 days had no influence on this increase. However, when Gln is reduced during the stress response, HSP70 expression is impaired. A reduction of Gln from 0.5 mM (physiological) to 0.125 mM (pathological) led to a 40% lower HSP70 level (P<0.002). In contrast, increasing Gln concentrations (up to 2 mM) had only minor stimulatory effects (about 15%). This Gln-dependency of heat mediated HSP70 expression was observed in resting as well as proliferating lymphocytes. Our data indicate that during periods of reduced plasma Gln levels the stress response of human lymphocytes is impaired. Thus, Gln may be essential to minimize the susceptibility of leucocytes to cytotoxic inflammatory mediators. This is a new aspect of the protective effect of Gln supplementation in critically ill patients. PMID- 11895152 TI - Pyruvate kinase type M2: a crossroad in the tumor metabolome. AB - Cell proliferation is a process that consumes large amounts of energy. A reduction in the nutrient supply can lead to cell death by ATP depletion, if cell proliferation is not limited. A key sensor for this regulation is the glycolytic enzyme pyruvate kinase, which determines whether glucose carbons are channelled to synthetic processes or used for glycolytic energy production. In unicellular organisms pyruvate kinase is regulated by ATP, ADP and AMP, by ribose 5-P, the precursor of the nucleic acid synthesis, and by the glycolytic intermediate fructose 1,6-P2 (FBP), thereby adapting cell proliferation to nutrient supply. The mammalian pyruvate kinase isoenzyme type M2 (M2-PK) displays the same kinetic properties as the pyruvate kinase enzyme from unicellular organisms. The mammalian M2-PK isoenzyme can switch between a less active dimeric form and a highly active tetrameric form which regulates the channeling of glucose carbons either to synthetic processes (dimeric form) or to glycolytic energy production (tetrameric form). Tumor cells are usually characterized by a high amount of the dimeric form leading to a strong accumulation of all glycolytic phosphometabolites above pyruvate kinase. The tetramer-dimer ratio is regulated by ATP, FBP and serine and by direct interactions with different oncoproteins (pp60v-src, HPV-16 E7). In solid tumors with sufficient oxygen supply pyruvate is supplied by glutaminolysis. Pyruvate produced in glycolysis and glutaminolysis is used for the synthesis of lactate, glutamate and fatty acids thereby releasing the hydrogen produced in the glycolytic glyceraldehyde 3-phosphate dehydrogenase reaction. PMID- 11895153 TI - Glutamine: essential for immune nutrition in the critically ill. AB - Critically ill patients on intensive care units are at an increased risk of sepsis, which is a major cause of mortality in these patients. Recent evidence suggests that impairment of the functioning of the immune system contributes to the development of sepsis in such patients. In particular, monocytes show reduced expression of HLA-DR antigen, associated with impaired antigen presenting capability and decreased phagocytic activity; lymphocytes show decreased proliferation in response to mitogens and T-helper cells show a shift in the Th1/Th2 ratio consistent with impaired immunity. The amino acid glutamine becomes conditionally essential in the critically ill, yet such patients frequently have a marked deficiency of glutamine; the reasons for this are still unclear. Glutamine is required by the cells of the immune system both as a primary fuel and as a carbon and nitrogen donor for nucleotide precursor synthesis. In vivo studies have demonstrated that glutamine is essential for optimal immune cell functioning for monocytes, lymphocytes and neutrophils. A number of trials of patients fed by the enteral or parenteral route have shown improved infectious morbidity when supplemented with glutamine. However, the exact mechanism of glutamine action in these patients remains to be determined. PMID- 11895154 TI - Fatty acids and lymphocyte functions. AB - The immune system acts to protect the host against pathogenic invaders. However, components of the immune system can become dysregulated such that their activities are directed against host tissues, so causing damage. Lymphocytes are involved in both the beneficial and detrimental effects of the immune system. Both the level of fat and the types of fatty acid present in the diet can affect lymphocyte functions. The fatty acid composition of lymphocytes, and other immune cells, is altered according to the fatty acid composition of the diet and this alters the capacity of those cells to produce eicosanoids, such as prostaglandin E2, which are involved in immunoregulation. A high fat diet can impair lymphocyte function. Cell culture and animal feeding studies indicate that oleic, linoleic, conjugated linoleic, gamma-linolenic, dihomo-gamma-linolenic, arachidonic, alpha linolenic, eicosapentaenoic and docosahexaenoic acids can all influence lymphocyte proliferation, the production of cytokines by lymphocytes, and natural killer cell activity. High intakes of some of these fatty acids are necessary to induce these effects. Among these fatty acids the long chain n-3 fatty acids, especially eicosapentaenoic acid, appear to be the most potent when included in the human diet. Although not all studies agree, it appears that fish oil, which contains eicosapentaenoic acid, down regulates the T-helper 1-type response which is associated with chronic inflammatory disease. There is evidence for beneficial effects of fish oil in such diseases; this evidence is strongest for rheumatoid arthritis. Since n-3 fatty acids also antagonise the production of inflammatory eicosanoid mediators from arachidonic acid, there is potential for benefit in asthma and related diseases. Recent evidence indicates that fish oil may be of benefit in some asthmatics but not others. PMID- 11895155 TI - Regulatory potential of n-3 fatty acids in immunological and inflammatory processes. AB - Over the last few years immunonutrition has gained increasing importance. Among other compounds lipids, especially n-3 polyunsaturated fatty acids, were shown to influence the immune response. The anti-inflammatory effects they exert can be induced by free fatty acids, triglyceride fatty acids, after incorporation into the membrane phopspholipid bilayer or following metabolism to eicosanoids. n-3 Fatty acids influence inflammatory cell activation processes from signal transduction to protein expression even involving effects at the genomic level. n 3 Fatty acid-mediated mechanisms decreased cytokine-induced adhesion molecule expression, thereby reducing inflammatory leucocyte-endothelium interactions and modified lipid mediator synthesis, thus influencing the transendothelial migration of leucocytes and leucocyte trafficking in general. Even the metabolic repertoire of specific immunocompetent cells such as cytokine release or proliferation is modified by n-3 fatty acids. Beyond this they regulate lipid homeostasis shifting the metabolic pathways towards energy supply thus optimizing the function of immune cells. Due to the regulatory impact on different processes of inflammatory and immune cell activation n-3 fatty acids provide positive effects on various states of immune deficiencies and diseases with a hyperinflammatory character, among which selected examples are presented. PMID- 11895156 TI - Parenteral nutrition with n-3 lipids in sepsis. AB - Dietary supplements of n-3 fatty acids have long been used to influence chronic inflammatory disorders. Recent studies with an immune-enhancing diet partly based on n-3 fatty acids report beneficial effects in patients with acute hyper inflammatory diseases, such as the sepsis syndrome or adult respiratory distress syndrome (ARDS). The possible suppression of exaggerated leucocyte activity, the improvement of microcirculatory events, as well as the opportunity to administer intravenous lipids enriched in n-3 fatty acids signal the possibility of a combination of parenteral caloric support and pharmacological intervention. Using parenteral administration of fish oil-based lipids, a new rapid and highly effective anti-inflammatory agent may allow the option to alter the immune status in hyper-inflammatory diseases such as sepsis and ARDS. PMID- 11895157 TI - n-3 fatty acids in psoriasis. AB - Increased concentrations of free arachidonic acid (AA) and its proinflammatory metabolites have been observed in psoriatic lesions. Replacement of arachidonic acid by alternative precursor polyunsaturated fatty acids (PUFA), especially eicosapentaenoic acid (EPA), which can be metabolized via the same enzymatic pathways as AA, might be a therapeutic option in psoriasis. However the results of studies evaluating the therapeutic benefit of dietary fish oil have been conflicting and not clearly dose-dependent. To overcome the slow kinetics and limited availability of oral supplementation, we have performed three studies to assess the efficacy and safety of an intravenously administered fish oil derived lipid emulsion on different forms of psoriasis. Patients received daily infusions of either an n-3 fatty acid-based lipid emulsion (Omegaven) or a conventional n-6 lipid emulsion (Lipoven) in different time and dose regimens. In addition to an overall assessment of the clinical course of psoriasis, EPA- and AA-derived neutrophil 5-lipoxygenase (LO)--products, thromboxane (TX) B2/B3, PAF and plasma free fatty acids were investigated. Treatment with n-3 fatty acids resulted in a considerably higher response rate than infusion of n-6 lipids. A more than 10 fold increase in neutrophil EPA-derived 5-LO product formation was noted in the n 3 group, accompanied by a rapid increase in plasma-free EPA within the first days. In conclusion, intravenous n-3-fatty acid administration causes reduction of psoriasis, which may be related to changes in inflammatory eicosanoid generation. The rapidity of the response to intravenous n-3 lipids exceeds by orders of magnitude the hitherto reported kinetics of improvement of psoriatic lesions upon use of oral supplementation. PMID- 11895158 TI - Immunomodulation by perioperative administration of n-3 fatty acids. AB - It has been increasingly reported that administration of n-3 fatty acids is beneficial in patients with inflammatory processes. This effect is most likely caused by different biological characteristics, including an immunomodulating effect of the products derived from n-3 fatty acids through eicosanoid metabolism. The aim of this study was to investigate the effect of perioperative administration of n-3 fatty acids on inflammatory and immune responses as well as on the postoperative course of patients with extended surgical interventions of the abdomen. In particular, the effect of n-3 fatty acids on interleukin-6 release and on granulocyte/monocyte function (HLA-DR expression) was studied. There was a downregulation of the inflammatory response, and, simultaneously, a smaller postoperative immune suppression in the n-3 fatty acid group. In addition, we observed shorter postoperative periods in the intensive care unit and on the regular medical wards as well as lower rates of severe infections. The results suggest that perioperative administration of n-3 fatty acids may have a favourable effect on outcome in patients with severe surgical interventions by lowering the magnitude of inflammatory response and by modulating the immune response. PMID- 11895159 TI - Glutamine supplementation in bone marrow transplantation. AB - An increasing number of clinical investigations have focused on supplementation of specialized enteral and parenteral nutrition with the amino acid glutamine. This interest derives from strong evidence in animal models and emerging clinical data on the efficacy of glutamine administration following chemotherapy, trauma, sepsis and other catabolic conditions. Glutamine has protein-anabolic effects in stressed patients and, among many key metabolic functions, is used as a major fuel/substrate by cells of the gastrointestinal epithelium and the immune system. These effects may be particularly advantageous in patients undergoing bone marrow transplantation (BMT), who exhibit post-transplant body protein wasting, gut mucosal injury and immunodeficiency. Studies to date indicate that enteral and parenteral glutamine supplementation is well tolerated and potentially efficacious after high-dose chemotherapy or BMT for cancer treatment. Although not all studies demonstrate benefits, sufficient positive data have been published to suggest that this nutrient should be considered as adjunctive metabolic support of some individuals undergoing marrow transplant. However, BMT is a rapidly evolving clinical procedure with regard to the conditioning and supportive protocols utilized. Thus, additional randomized, double-blind, controlled clinical trials are indicated to define the efficacy of glutamine with current BMT regimens. PMID- 11895160 TI - Impact of n-3 fatty acid supplemented parenteral nutrition on haemostasis patterns after major abdominal surgery. AB - In various diseases n-3 fatty acids exert anti-inflammatory properties. These effects seem to be related to the uptake and incorporation of eicosapentaenoic acid (EPA) into the cellular substrate pool after dietary intake of EPA, which is contained in fish oils (FO). In the state of inflammation EPA is released to compete with arachidonic acid (AA) for metabolism at the cyclo-oxygenase and the 5-lipoxygenase level. The metabolites of EPA have less inflammatory and chemotactic potency than the substances derived from AA. In addition to positive effects, early studies pointed towards prolonged bleeding times after dietary intake of n-3 fatty acids. This study was undertaken to address the issue of potential coagulation disturbances associated with postoperative parenteral FO administration. This was a prospective, randomised, double blinded clinical trial, carried out in two operative intensive care units (13 and 16 beds) in a university hospital. Forty-four patients undergoing elective major abdominal surgery participated in the trial. Patients were randomly assigned to receive total parenteral nutrition (TPN) supplemented with either soybean oil (SO, Lipovenoess 10% PLR; 1.0 g/kgBW per day; n = 20) for five days or with a combination of FO and SO (FO, Omegaven; 0.2 g/kgBW per day plus SO, Lipovenoes 10% PLR; 0.8 g/kgBW per day, n = 24), respectively. Blood samples were taken preoperatively (day -1), prior to (day 1) during (days 2-5) and after TPN (day 6). The coagulation parameters thromboplastin time (Quick), activated partial thromboplastin time (aPTT), fibrinogen and antithrombin III were measured. To differentially assess activation levels of extrinsic and intrinsic coagulation pathway, factors VIIa and XIIa were quantified. Moreover platelet function was determined by resonance thrombography. Baseline values of coagulation and platelet function were comparable in both groups, but coagulation activity dropped after surgery. Over the observation period of 6 days, however, physiological levels were regained. No clinically significant differences were observed between the SO- and SO + FO- group. These findings suggest that infusion of fish oil in doses up to 0.2 g/kgBW per day is safe regarding coagulation and platelet function. PMID- 11895161 TI - Body-weight changes are clearly reflected in plasma concentrations of leptin in female mink (Mustela vison). AB - The mink is a seasonal breeder with a propensity for seasonal fatness, and it is very responsive to changes in energy supply. The objectives of the present study were first, to validate a multi-species leptin assay for mink (Mustela vison) plasma, and second, to evaluate how plasma leptin and insulin concentrations responded to energy restriction and body-weight loss and refeeding with restoration of body reserves. The study was performed with six very fat yearling females (initial mean body weight 1,451 (SD 119) g, i.e. approximately 300 g more than for a female in normal body condition). The animals were fed in restricted amounts (about 35 % metabolizable energy requirement for maintenance) in order to reach a very lean body condition. The target weight of 800 g was reached after about 1 month of restriction. The animals were then refed ad libitum until almost complete weight recovery. Blood samples were taken on days 1, 24, 34 (end of restriction), 44, 55 and 71 (end of experiment) and analysed for plasma concentrations of leptin and insulin. Three females were mated on day 44. Leptin and insulin concentrations mirrored each other and clearly reflected changes in body weight. Significant (P<0.001) Pearson correlation coefficients of 0.75 (leptin-insulin), 0.72 (leptin-body weight) and 0.59 (insulin-body weight) were found. Two of the three females that were mated gave birth to normal litters. It was concluded that the leptin assay yielded acceptable results for animals with body weight:fat content within the range investigated here, and that plasma leptin reflected body fat mass. PMID- 11895162 TI - Increased phagocytosis and production of reactive oxygen species by neutrophils during magnesium deficiency in rats and inhibition by high magnesium concentration. AB - Recent studies underline the importance of the immunoinflammatory processes in the pathology of Mg deficiency. Neutrophils possess a superoxide anion-generating NADPH oxidase and its inappropriate activation may result in tissue damage. The aim of the present study was to assess the effect of experimental Mg deficiency in the rat on polymorphonuclear leucocytes (PMN) activity and the role of increasing extracellular Mg. Weaning male Wistar rats were fed either a Mg deficient or a control diet for 8 d. In Mg-deficient rats, the characteristic inflammatory response was accompanied by a marked increase in the number of PMN. Higher plasma interleukin 6 and NO concentrations and increased lipid peroxidation in the heart were found in Mg-deficient rats as compared with control rats. As shown by chemiluminescence studies, basal neutrophil activity from Mg-deficient rats was significantly elevated when compared with neutrophils from control rats. Moreover, the chemiluminescence of PMN from Mg-deficient rats was significantly higher than that of control rats following phorbol myristate acetate or opsonized zymosan activation. PMN from Mg-deficient rats also showed an increased activity of phagocytosis in comparison with neutrophils from control animals. Increasing extracellular Mg concentration in the incubating medium of PMN (0.8 v. 8.0 mM) decreased the chemiluminescence activity of PMN from control rats following opsonized zymosan activation. Chemiluminescence activities of PMN from Mg-deficient rats following phorbol myristate acetate or opsonized zymosan challenge were also decreased by high extracellular Mg concentration. From this work, it appears that PMN activation is an early consequence of Mg deficiency and that high extracellular Mg concentration inhibits free radicals generation. PMID- 11895163 TI - Niacin (nicotinic acid) in non-physiological doses causes hyperhomocysteineaemia in Sprague-Dawley rats. AB - Niacin (nicotinic acid) in its non-physiological dose level is known to be an effective lipid-lowering agent; its potential risk as a therapeutic agent, however, has not been critically considered. Since niacin is excreted predominantly as methylated pyridones, requiring methionine as a methyl donor, the present study was undertaken to examine whether metabolism of the amino acid is altered in the presence of large doses of niacin. Male Sprague-Dawley rats were given a nutritionally adequate, semi-synthetic diet containing niacin at a level of either 400 or 1000mg/kg diet (compared to 30mg/kg in the control diet) for up to 3 months. Supplementation with niacin (1,000 mg/kg diet) for 3 months resulted in a significant increase in plasma and urinary total homocysteine levels; this increase was further accentuated in the presence of a high methionine diet. The hyperhomocysteineaemia was accompanied by a significant decrease in plasma concentrations of vitamins B6 and B12, which are cofactors for the metabolism of homocysteine. The homocysteine-raising action of niacin, in particular, has an important toxicological implication, as hyperhomocysteineaemia is considered to be an independent risk factor for arterial occlusive disease. The niacin-associated change in homocysteine status may be an important limiting factor in the use of this vitamin as a lipid-lowering agent. PMID- 11895164 TI - Interrelationships between dairy product intake, microflora metabolism, faecal properties and plasmid dissemination in gnotobiotic mice. AB - We previously described the effects of intake of dairy products on plasmid dissemination in the digestive tract of gnotobiotic mice associated with human faecal flora (HFF) and found that yoghurt, heat-treated yoghurt (HTY) and milk reduced population levels of transconjugants compared with findings in mice fed a standard mouse diet. In the case of lactose intake, transconjugants were not detected. The aim of the present study was to assess the possible interrelationships between these observations and other variables (bacterial ecology, pH, moisture, enzyme activities, short-chain fatty acid (SCFA) contents, lactic acid contents). Much of the interest of the present comparison lies in the fact that the animals were homogeneous in terms of age, gender, food and intestinal microflora, owing to the gnotobiotic mouse model maintained in sterile isolators. We observed no variation in SCFA and lactic acid contents or in the population levels of strictly anaerobic strains of Bacteroides and Bifidobacterium, and of the facultative anaerobic recipient Escherichia coli PG1 strain. The main modifications were the reduction of population levels of transconjugants in mice receiving yoghurt, HTY and milk, and concomitantly an increase of beta-galactosidase and a decrease of beta-glucosidase activities, compared with control mice fed a standard diet. Total inhibition of plasmid transfer was observed in HFF mice consuming lactose, and concomitantly the two enzyme activities (beta-glucosidase and beta-galactosidase) were increased, compared with the findings in control mice fed a standard diet. In axenic mice consuming lactose, plasmid transfer occurred, beta-galactosidase was not detected and beta-glucosidase was decreased. It is therefore proposed that these two enzyme activities influence plasmid transfer and persistence of transconjugants in the digestive tract of HFF associated mice. When both activities were increased there was a total inhibition of plasmid transfer (case of lactose intake). When beta-galactosidase increased and beta-glucosidase decreased (case of yoghurt, HTY and milk), plasmid transfer occurred at a lower efficiency than in the control group, resulting in lower population levels of transconjugants. PMID- 11895165 TI - Net energy value of two low-digestible carbohydrates, Lycasin HBC and the hydrogenated polysaccharide fraction of Lycasin HBC in healthy human subjects and their impact on nutrient digestive utilization. AB - The metabolizable energy content of low-digestible carbohydrates does not correspond with their true energy value. The aim of the present study was to determine the tolerance and effects of two polyols on digestion and energy expenditure in healthy men, as well as their digestible, metabolizable and net energy values. Nine healthy men were fed for 32 d periods a maintenance diet supplemented either with dextrose, Lycasin HBC (Roquette Freres, Lestrem, France), or the hydrogenated polysaccharide fraction of Lycasin HBC, at a level of 100 g DM/d in six equal doses per d according to a 3 x 3 Latin square design with three repetitions. After a 20 d progressive adaptation period, food intake was determined for 12d using the duplicate meal method and faeces and urine were collected for 10 d for further analyses. Subjects spent 36 h in one of two open circuit whole-body calorimeters with measurements during the last 24h. Ingestion of the polyols did not cause severe digestive disorders, except excessive gas emission, and flatulence and gurgling in some subjects. The polyols induced significant increases in wet (+45 and +66% respectively, P<0.01) and dry (+53 and +75 % respectively, P<0.002) stool weight, resulting in a 2% decrease in dietary energy digestibility (P<0.001). They resulted also in significant increases in sleeping (+4.1%, P<0.03) and daily energy expenditure (+2.7 and +2.9% respectively, P<0.02) compared with dextrose ingestion. The apparent energy digestibility of the two polyols was 0.82 and 0.79 respectively, their metabolizable energy value averaged 14.1 kJ/g DM, and their net energy value averaged 10.8 kJ/g DM, that is, 35 % less than those of sucrose and starch. PMID- 11895166 TI - Acute effects of ingestion of black tea on postprandial platelet aggregation in human subjects. AB - Results of population studies suggest that black tea can reduce cardiovascular risk. Effects of black-tea polyphenols to reduce platelet aggregability may help to explain any benefits. Given that black tea is often consumed with and after meals, and man spends much of his life in the postprandial state, the objective of the present study was to investigate the acute effects of ingestion of black tea on postprandial platelet aggregation ex vivo. Twenty healthy participants had platelet aggregation and blood lipids assessed before and 4 h after the ingestion of 50 g dairy fat on two occasions in random order, corresponding to black tea or hot water. Black tea or hot water (one cup) was consumed immediately following the dairy fat, then after 15 and 30 h. Platelet aggregation ex vivo was assessed in platelet-rich plasma in response to three concentrations of collagen (0.2, 0.6, 20 microg/ml) and ADP (2, 4, 8 microM). Urinary concentrations of 4-O methylgallic acid were used as an indicator that tea polyphenols were absorbed. Serum total cholesterol and triacylglycerol concentrations increased significantly 4 h after ingesting the dairy fat, but there was no significant difference between black tea and hot-water treatments on the cholesterol or triacylglycerol responses. Urinary 4-O-methylgallic acid concentrations were significantly increased following ingestion of black tea (P=0.0001) but not water. Black tea in comparison to hot water did not inhibit collagen or ADP induced postprandial platelet aggregation. The results of this study do not support the suggestion that reduced postprandial platelet aggregability contributes to any benefits of black tea on cardiovascular risk. PMID- 11895167 TI - Pancreatic islet insulin secretion and metabolism in adult rats malnourished during neonatal life. AB - Pancreatic islets were isolated from rats that had been nursed by dams fed with a control or an 8.7% protein diet during the first 12 d of the lactation period. Glucose-induced insulin secretion from islets in the 8.7% protein group was reduced 50%. The islet insulin and DNA content were similar, whereas the pancreatic insulin content was reduced by 30 % in the rats fed 8.7 % protein. In order to elucidate the mechanism responsible for the attenuation of insulin secretion, measurements were performed of the activity of several islet enzymes that had previously been supposed to be involved in the coupling of glucose stimulation to insulin secretion. Islet glucose oxidation was unaffected, but glucose-stimulated hydrolysis of phosphatidylinositol was reduced by one-third in the islets of rats fed 8.7% protein. The activity of mitochondrial glycerophosphate dehydrogenase was similar in islets of rats fed the 8.7% protein diet and those fed the control diet. The activity of Ca-independent phospholipase A2 was increased fourfold in the islets of rats fed 8.7% protein. It is concluded that impairment of glucose-induced insulin secretion in rats fed a low-protein diet may be caused by attenuation of islet phosphatidylinositol hydrolysis, and it is tentatively suggested that the increased activity of Ca-independent phospholipase A2 in islets of rats fed a low-protein diet may participate in the stimulation of apoptosis. PMID- 11895168 TI - Effect of eicosapentaenoic acid ethyl ester v. oleic acid-rich safflower oil on insulin resistance in type 2 diabetic model rats with hypertriacylglycerolaemia. AB - The purpose of the present study was to test whether hyperlipidaemia and insulin resistance in type 2 diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats can be improved by dietary supplementation with purified eicosapentaenoic acid (EPA) or oleic acid (OA). Male OLETF rats were fed powdered chow (510 g fat/kg) alone (n 8) or chow supplemented with 10 g EPA- (n 8) or OA- (n 8) rich oil/kg per d from 5 weeks until 30 weeks of age. An oral glucose tolerance test and hyperinsulinaemic euglycaemic clamp was performed at 25 and 30 weeks of age. EPA supplementation resulted in significantly (P<0.05) reduced plasma lipids, hepatic triacylglycerols, and abdominal fat deposits, and more efficient in vivo glucose disposal compared with OA supplementation and no supplementation. OA supplementation was associated with significantly increased insulin response to oral glucose compared with EPA supplementation and no supplementation. Inverse correlation was noted between glucose uptake and plasma triacylglycerol levels (r -086, P<0.001) and abdominal fat volume (r -0.80, P<0.001). The result of oral glucose tolerance test study showed that the rats fed EPA tended to improve glucose intolerance, although this was not statistically significant. Levels of plasma insulin at 60 min after glucose was significantly increased in rats fed OA compared with the other two groups. The results indicate that long-term feeding of EPA might be effective in preventing insulin resistance in diabetes-prone rats, at least in part, due to improving hypertriacylglycerolaemia. PMID- 11895169 TI - Inconsistencies in bioelectrical impedance and anthropometric measurements of fat mass in a field study of prepubertal children. AB - The present study examined the consistency of bioelectrical impedance analysis (BIA) and anthropometric measurements in body composition analysis in a field study of prepubertal children using a representative group of 2286 5-7-year-old children from Kiel, north-west Germany. Body composition was assessed using anthropometric measures (A; four skinfolds) and BIA. Various published algorithms (according to Lohmann (1986) and Deurenberg et al. (1990) for A, Kushner et al. (1992), Schaefer et al. (1994) and Wabitsch et al. (1996) for BIA and Goran et al. (1996) for a combined approach) were used to estimate body composition. Using A resulted in a sum of four skinfolds varying between age-dependent median values of 24.0 and 28.2 mm in boys and 30.5 and 33.3 mm in girls. When fat mass (FM) was calculated from A, age- and algorithm-dependent differences in median values were observed, with values varying between 8.5 and 14.6% for boys and 1.11 and 14.9% for girls. Using different algorithms (Lohmann (1986) v. Deurenberg et al. (1990)) only minor inconsistencies were observed. BIA-derived resistance index (height2/resistance) varied between 18.8 and 24.4 cm2/omh for boys and 17.1 and 19.0cm2/ohm for girls. Using four different algorithms to estimate FM from BIA data resulted in high intra-individual variances in percentage FM (from 13.8 to 33.4) as well as in the prevalence of overweight (from 14.7 to 98.4% for boys and from 42.3 to 98.5 % for girls). Data obtained using the different BIA algorithms showed some, or even marked, inconsistencies as well as systematic deviations (an overestimation of FM at low percentage FM, Schaefer et al. (1994) v. Wabitsch et al. (1996)). When comparing BIA with A, BIA systematically overestimated FM. The differences between the results were influenced by BMI, gender and height. Considerable inconsistencies were observed at low BMI (<10th percentile) for girls and for small children. Although the within-observer as well as between observer CV for both techniques are acceptable, we recommend caution in relation to the algorithms used for data analysis. The use of an interchange table of percentage FM derived from different algorithms for different percentile groups of skinfold thicknesses is recommended. PMID- 11895170 TI - Anthropometric measurements in the elderly: age and gender differences. AB - In clinical practice and epidemiological surveys, anthropometric measurements represent an important component of nutritional assessment in the elderly. The anthropometric standards derived from adult populations may not be appropriate for the elderly because of body composition changes occurring during ageing. Specific anthropometric reference data for the elderly are necessary. In the present study we investigated anthropometric characteristics and their relationship to gender and age in a cross-sectional sample of 3,356 subjects, randomly selected from an elderly Italian population. In both sexes, weight and height significantly decreased with age while knee height did not. The BMI was significantly higher in women than in men (27.6 SD 5.7 v. 26.4 SD 3.7; P<0.001) and it was lower in the oldest than in the youngest subjects (P<0.05) of both genders. The 75th year of age was a turning point for BMI as for other anthropometric measurements. According to BMI values, the prevalence of malnutrition was lower than 5 % in both genders, whereas obesity was shown to have a higher prevalence in women than in men (28% v. 16%; P<0.001). Waist circumference and waist: hip ratio values were higher for the youngest men than for the oldest men (P<0.05), whereas in women the waist: hip ratio values were higher in the oldest women, suggesting that visceral redistribution in old age predominantly affects females. In conclusion, in the elderly the oldest subjects showed a thinner body frame than the youngest of both genders, and there was a more marked fat redistribution in women. PMID- 11895171 TI - Minor importance of de novo lipogenesis on energy expenditure in human. PMID- 11895172 TI - Decreased expression of the vitamin C transporter SVCT1 by ascorbic acid in a human intestinal epithelial cell line. AB - Vitamin C (ascorbic acid) is an essential nutrient that is involved in a number of cellular processes. However, unlike most mammals, man is unable to synthesize vitamin C and it must therefore be acquired from the diet. Absorption of vitamin C is achieved by two transporters, SVCTI and SVCT2, recently cloned from rat and human kidney. SVCT1 is thought to be the predominant transporter in the intestine. Vitamin C supplements are increasingly common, thus contributing to an increased dietary load, and therefore the aim of the present study was to investigate the effect of high doses of ascorbic acid on SVCT1 expression. Using the Caco-2 TC7 cell model of small intestinal enterocytes, we measured the effects of ascorbic acid (4.5 mg/ml culture medium) on L-[14C]ascorbic acid uptake and SVCT1 expression (determined by reverse transcription-polymerase chain reaction). Ascorbic acid uptake was decreased significantly in Caco-2 TC7 cells exposed to ascorbate for 24 h (-50%, P<0.0005). Expression of SVCT1 was also significantly reduced by exposure to elevated levels of ascorbate for 24h (-77%, P<0.005). Taken together these results suggest that high-dose supplements might not be the most efficient way of increasing the body pool of vitamin C. PMID- 11895173 TI - Learning power of single behavioral units in acquisition of a complex spatial behavior: an observational learning study in cerebellar-lesioned rats. AB - By combining an observational spatial learning paradigm with a cerebellar lesion that blocks the acquisition of new spatial strategies, it is possible to separate a complex spatial behavior into its fundamental units to study which relationships among units have to be maintained so that the entire behavior might be acquired. Normal rats were first allowed to observe demonstrator rats performing single explorative behaviors (circling, extended searching, direct finding), then were hemicerebellectomized and, finally, tested in the Morris water maze. In spite of the cerebellar lesion, the observer rats displayed exploration abilities that closely matched the previously observed behaviors. These results indicate that the single facets that form the strategy repertoire can be independently acquired. PMID- 11895174 TI - Sensitivity to the locomotor stimulant effects of ethanol and allopregnanolone is influenced by common genes. AB - Allopregnanolone is a neuroactive steroid that, like ethanol (EtOH), has stimulant, anxiolytic, ataxic, and depressant effects. Two experiments tested the hypothesis that sensitivity to the locomotor stimulant effects of these drugs is influenced by a common set of genes. Sensitivity to the locomotor stimulant effects of allopregnanolone was determined in 24 BXD recombinant inbred (RI) strains. Strain means were positively correlated with extant means for EtOH stimulation in 20 of the same strains. Quantitative trait locus (QTL) analysis provisionally identified many loci, including several known to influence sensitivity to EtOH. Sensitivity to allopregnanolone was also measured in FAST and SLOW mice, which were selectively bred for differential locomotor response to EtOH, to determine whether selection has also altered allopregnanolone sensitivity. FAST mice were more sensitive to the stimulant effects of allopregnanolone compared with SLOW mice. These data suggest that sensitivity to the locomotor stimulant effects of these drugs is influenced by common genes. PMID- 11895175 TI - Glutamatergic transmission in the nucleus of the solitary tract modulates memory through influences on amygdala noradrenergic systems. AB - The authors examined whether glutamate release from the vagus nerve onto the nucleus of the solitary tract (NTS) is one mechanism by which the vagus influences memory and neural activity in limbic structures. Rats trained to drink from a spout were given a footshock (0.35 mA) on Day 5 after approaching the spout. Phosphate-buffered saline or 5.0, 50.0, or 100.0 nmol/0.5 microl glutamate was then infused into the NTS. Glutamate (5.0 or 50.0 nmol) significantly enhanced memory on the retention test. In Experiment 2, this effect was attenuated by blocking noradrenergic receptors in the amygdala with propranolol (0.3 microg/0.5 microl). Experiment 3 used in vivo microdialysis to determine whether footshock plus glutamate (50.0 nmol) alters noradrenergic output in the amygdala. These treatments caused a significant and long-lasting increase in amygdala noradrenergic concentrations. The results indicate that glutamate may be one transmitter that conveys the effects of vagal activation on brain systems that process memory. PMID- 11895176 TI - Ethanol-induced conditioned taste aversion in 15 inbred mouse strains. AB - This study used a genetic correlational strategy to characterize the neurobiological basis of ethanol's (0, 2, or 4 g/kg) aversive effects as indexed by conditioned taste aversion. Substantial strain differences in taste aversion and hypothermia were observed, but the genetic correlation between these phenotypes was not significant. However, significant genetic correlations were observed between taste aversion and ethanol-related behaviors measured in previous studies, including home-cage ethanol preference (r = .68) and ethanol withdrawal severity (r = -.69). Strains showing stronger taste aversion tended to show lower ethanol preference and higher withdrawal severity. This pattern of findings is consistent with previous studies suggesting a commonality in neurobiological mechanisms underlying these phenotypes. These results do not support the hypothesis that ethanol-induced taste aversion is mediated by the drug's rewarding properties. PMID- 11895177 TI - The effect of amiloride on operantly conditioned performance in an NaCl taste detection task and NaCl preference in C57BL/6J mice. AB - A 2-response operant taste discrimination procedure, modified to assess taste sensitivity in water-restricted C57BL/6J mice, revealed a detection threshold of 0.065 M sodium chloride. Amiloride increased the threshold by approximately 1 log10 unit. These results are the first to demonstrate the necessity of the amiloride-sensitive taste transduction pathway in the normal detection of low concentrations of sodium chloride in mice and provide a functional context in which to evaluate electrophysiological findings. Two-bottle preference tests performed with these mice and additional naive mice revealed only marginal, if any, effects of amiloride on salt intake behavior, highlighting the importance of considering the relative attributes and limitations of different behavioral assays of taste function. PMID- 11895178 TI - Renewal of drug seeking by contextual cues after prolonged extinction in rats. AB - Contextual stimuli associated with drug exposure can modulate various effects of drugs, but little is known about their role in relapse to drug seeking. Using a renewal procedure, the authors report that drug-associated contextual stimuli play a critical role in relapse to drug-seeking previously maintained by a heroin cocaine mixture (speedball). Rats were trained to self-administer speedball, after which drug-reinforced behavior was extinguished over 20 days in the self administration context or in a different context. On the test day, rats exposed to the drug-associated context, after extinction in a different context, reliably renewed drug seeking. The authors suggest that the renewal procedure can be used to study mechanisms underlying relapse to drug seeking elicited by drug associated contextual stimuli. PMID- 11895179 TI - Reinforcer devaluation abolishes conditioned cue preference: evidence for stimulus-stimulus associations. AB - In the conditioned cue preference (CCP) task, the subject is presented with a cue paired with food reward, resulting in a preference for the paired cue when allowed to choose later. To clarify the learning involved, the authors devalued the reinforcer after training by inducing a taste aversion to the food. In five 30-min sessions, rats were confined in 1 arm of a radial arm maze and presented with food. These reinforced sessions alternated with 5 unreinforced sessions in a nonadjacent arm. Devaluation was then accomplished in 1 group by inducing taste aversion; controls received either saline or unpaired lithium chloride treatment. When tested later, both the saline group and the unpaired group preferred the previously reinforced arm, but the devalued group showed aversion to it. Thus, CCP is mediated by the stimulus-reinforcer association; when the reinforcer is devalued, the preference is also abolished. PMID- 11895180 TI - Neuronal correlates of conditioned inhibition of the eyeblink response in the anterior interpositus nucleus. AB - Conditioned inhibition (CI) of the rat eyeblink response and the neuronal correlates of CI in the cerebellar interpositus nucleus were examined in the present study. In Experiment 1, CI was established with a novel, 3-group design. In Experiment 2, neuronal activity in the anterior interpositus nucleus was recorded during CI training and testing. Each rat was given 2 training phases and then tested for CI with summation and retardation tests. Rats given CI training showed behavioral inhibition compared with rats in 2 control groups. Neuronal activity in the anterior interpositus nucleus correlated with behavioral responding during discrimination training and during the summation test. The results suggest that neurons in the cerebellar anterior interpositus nucleus may participate in the acquisition or expression of CI. PMID- 11895181 TI - Medial prefrontal cortex and pavlovian conditioning: trace versus delay conditioning. AB - Pavlovian eyeblink (EB) conditioning was studied in both trace and delay paradigms in rabbits (Oryctolagus cuniculus) with either medial prefrontal cortex (mPFC) lesions or sham lesions. mPFC lesions of prelimbic cortex (Brodmann's Area 32) retarded EB conditioning in the trace but not the delay paradigm. However, this effect was significant only when the conditioned stimulus (CS) was 500 rather than 100 ms in duration. Lesions of the anterior cingulate cortex (Area 24) did not affect EB conditioning in a trace paradigm. Accompanying CS-evoked heart rate slowing was attenuated under all conditions by the mPFC lesions, although this result was not always statistically significant. PMID- 11895182 TI - Olfactory-mediated fear-potentiated startle. AB - Recently, R. Richardson, A. Vishney, and J. Lee (1999) reported that ambient odor cues that were previously paired with footshock potentiate the acoustic startle response in rats. The authors of the present study extend those findings by using a discrete 4-s amyl acetate odor paired with footshock to address several parametric issues that might be important for using odorants as conditioned stimuli (CSs) in this paradigm. Amyl acetate (5%) had no significant effect on startle in untrained rats but did potentiate startle in rats that received 1, 2, 5, or 10 odor-shock pairings. Fear-potentiated startle decreased but was still significant up to 40 days after conditioning and could be measured in test trials separated by as little as 30 s. The magnitude of potentiated startle decreased with decreasing concentrations of amyl acetate (5%-5 x 10-9%). The anxiolytic compound buspirone (10 mg/kg) significantly attenuated olfactory-mediated fear potentiated startle. PMID- 11895183 TI - A connectionist model of septohippocampal dynamics during conditioning: closing the loop. AB - Septohippocampal interactions determine how stimuli are encoded during conditioning. This study extends a previous neurocomputational model of corticohippocampal processing to incorporate hippocamposeptal feedback and examines how the presence or absence of such feedback affects learning in the model. The effects of septal modulation in conditioning were simulated by dynamically adjusting the hippocampal learning rate on the basis of how well the hippocampal system encoded stimuli. The model successfully accounts for changes in behavior and septohippocampal activity observed in studies of the acquisition, retention, and generalization of conditioned responses and accounts for the effects of septal disruption on conditioning. The model provides a computational, neurally based synthesis of prior learning theories that predicts changes in medial septal activity based on the novelty of stimulus events. PMID- 11895185 TI - Effects of hippocampal N-methyl-D-aspartate infusion on locomotor activity and prepulse inhibition: differences between the dorsal and ventral hippocampus. AB - Prepulse inhibition (PPI) of the acoustic startle response and open-field locomotor activity were measured after bilateral infusion of N-methyl-D-aspartate into the ventral (0.10, 0.25, 0.50 microg/side) and dorsal (0.10, 0.25, 0.50, 0.70 microg/side) hippocampus of Wistar rats. Dose-dependent hyperactivity and disruption of PPI--behavioral effects related to psychotic symptoms--were observed after ventral infusions but were virtually absent after dorsal infusions. This functional dorsal-ventral difference might be related to the different connections of the dorsal and ventral hippocampus with the amygdala, nucleus accumbens, and prefrontal cortex, which have been implicated in the regulation of locomotor activity and PPI. Hippocampal overactivity has been associated with schizophrenia. The findings suggest that overstimulation of the ventral hippocampal projections may contribute to behavioral outcomes related to psychotic symptoms. PMID- 11895186 TI - Extensive cytotoxic lesions of the rat retrosplenial cortex reveal consistent deficits on tasks that tax allocentric spatial memory. AB - Despite the connections of the retrosplenial cortex strongly suggesting a role in spatial memory, the lesion data to date have been equivocal. Whether subjects are impaired after retrosplenial lesions seems to depend on whether the lesions were aspirative or excitotoxic, with the latter failing to produce an impairment. A shortcoming of previous excitotoxic lesion studies is that they spared the most caudal part of the retrosplenial cortex. The present study thus used rats with extensive neurotoxic lesions of the retrosplenial cortex that encompassed the entire rostrocaudal extent of this region. These rats were consistently impaired on several tests that tax allocentric memory. In contrast, they were unimpaired on an egocentric discrimination task. Although the lesions did not appear to affect object recognition, clear deficits were found for an object-in-place discrimination. The present study not only demonstrates a role for the retrosplenial cortex in allocentric spatial memory, but also explains why previous excitotoxic lesions have failed to detect any deficits. PMID- 11895184 TI - Role of the rodent hippocampus in paired-associate learning involving associations between a stimulus and a spatial location. AB - The ability of rats with control or hippocampal lesions to learn an object-place, odor-place, or object-odor paired-associate task was assessed in a cheeseboard maze apparatus. The data indicate that rats with hippocampal lesions were significantly impaired, compared with controls, in learning both the object-place and the odor-place paired-associate tasks. However, rats with hippocampal lesions learned the object-odor paired-associate task as readily as did controls. The data suggest that the rodent hippocampus is involved in paired-associate learning when a stimulus must be associated with a spatial location. However, the hippocampus is not involved in paired-associate learning when the association does not involve a spatial component. PMID- 11895187 TI - The entorhinal cortex-nucleus accumbens pathway and latent inhibition: a behavioral and neurochemical study in rats. AB - Latent inhibition (LI) refers to the decrease in conditioned response produced by the repeated nonrein-forced preexposure to the to-be-conditioned stimulus. Experiment I investigated the effects of electrolytic lesions of the entorhinal cortex on LI in a conditioned emotional response procedure. Entorhinal cortex lesions attenuated LI. Experiments 2 and 3 investigated whether this attenuation of LI could result from a modification in nucleus accumbens (NAcc) dopamine (DA) release. Rats with entorhinal cortex lesions displayed normal spontaneous and amphetamine-induced locomotor activity, as well as normal basal and amphetamine induced release of DA within the NAcc (assessed by microdialysis). Taken together, these results show that entorhinal cortex lesions disrupt LI in a way that is unlikely to be due to an alteration of DA release within the NAcc. PMID- 11895188 TI - Study of eosinophil activation in nasal mucosa in patients with perennial nasal allergy: effects of CO2 laser surgery. AB - Carbon dioxide (CO2) laser surgery has been shown to be clinically effective in the treatment of nasal allergy. To investigate the mechanisms of eosinophil infiltration and activation underlying the therapeutic effects of CO2 laser surgery, we examined changes in the cytological profile of nasal mucosa after surgery. Twenty-two patients with perennial nasal allergy against house-dust mites underwent two or three rounds of laser surgery at 1-month intervals on an outpatient basis. The following parameters were evaluated at each visit: (i) improvement of clinical symptoms (nasal obstruction, rhinorrhea, and sneezing), (ii) percentage of infiltrating eosinophils in nasal mucosa, and (iii) the degree of EG2+ cells and intracellular adhesion molecule 1 (ICAM-1) expression by immunocytochemistry. All clinical symptoms significantly decreased after surgery. Significant reductions in eosinophil infiltration (p < 0.01) and the percentage of EG2+ cells (p < .005) were observed also. However, the degree of ICAM-1 expression in epithelial cells was not changed. These results suggest that CO2 laser surgery partially reduced the allergic reactions, leading to improvement of clinical symptoms. PMID- 11895189 TI - Chromametric assessment of nasal mucosal color and its application in patients with nasal allergy. AB - The color of the nasal mucosa was evaluated quantitatively in patients with nasal allergy and was compared with that in normal subjects. The chromaticity of the inferior turbinate was measured with the aid of a chromometer coupled with a light projection unit. Chromaticity was indicated by two parameters: the x and y values, which indicate the proportions of the red and green components of a color, respectively. Subjects included patients with Japanese cedar pollinosis (CP; n = 70) and perennial allergic rhinitis (PAR; n = 49), and normal subjects (n = 60). Mucosal blood flow in the inferior turbinate also was measured with a laser Doppler flowmeter in normal subjects. In normal subjects, the x value ranged between 0.4013 and 0.4539, and they value ranged between 0.3065 and 0.3379. The averages of the x and y values were 0.4264 +/- 0.0119 and 0.3204 +/- 0.0067, respectively. Age and sex were not related to the chromaticity value. Application of 0.1% epinephrine (0.12 mL) by aerosol spray decreased the x value significantly, in association with a significant decrease in blood flow in normal subjects. The x value of chromaticity in PAR patients was significantly lower than that in normal subjects; this showed the reduced red component of the color. There was no significant difference in the y value between PAR patients and the normal group. In the CP group, the x and y values were not significantly different from values in normal subjects. However, the x value in the first week after the onset of symptoms was significantly greater than that in the normal group. It subsequently decreased to the normal level in the second week and fell to a significantly lower level thereafter. This study found that the chromaticity of the nasal mucosa reflects local blood flow and that the circulatory dynamics in CP patients changes with time luring the pollinosis season. PMID- 11895190 TI - Endoscopic management of cerebrospinal fluid leaks. AB - To examine the diagnosis and treatment of patients with cerebrospinal fluid (CSF) rhinorrhea, a cohort of 36 patients treated between 1993 and 2000 were examined to consider the role of imaging and other diagnostic tests such as the presence of beta-transferrin in nasal secretion. The etiology of the condition was considered and was found to be congenital in 7 patients and acquired in 29 patients of which in 15 patients it was traumatic, in 12 patients it occurred spontaneously, and in 2 patients it was associated with Wegener's granulomatosis. Where it was possible to obtain nasal secretion, beta-transferrin proved a highly sensitive and specific test and imaging included computed tomography (CT), CT cisternography, and magnetic resonance imaging (MRI) of which fine detail coronal CT and MRI proved the most helpful. In six patients neither imaging nor beta transferrin could be used to confirm the diagnosis in which case intrathecal fluorescein was used. Repair was performed endoscopically in all cases with one exception where the defect was felt to be too large for this technique. Middle turbinate mucosa, cartilage, and fascia were the preferred repair materials in the anterior skull base whereas dermalfat was preferentially used in the sphenoid. The overall success rate for an endoscopic approach was 94% although in three cases a second endoscopic procedure was required to produce closure and external approaches were used in two additional patients. The use of a diagnostic algorithm is helpful in both confirming the presence of CSF rhinorrhea and the optimum approach. In the vast majority of cases an endoscopic repair will be successful and it avoids many of the complications associated with craniotomy, particularly in a young population. Therefore, it is our preferred option, although surgeons must be prepared for alternative procedures should these prove necessary. PMID- 11895191 TI - Topical antibiotic, antifungal, and antiseptic solutions decrease ciliary activity in nasal respiratory cells. AB - This study was designed to investigate whether topical solutions, as they are used in the treatment of selected cases of rhinosinusitis, influence nasal mucociliary clearance. The objective of this study was to evaluate the effects of the following topical solutions on the ciliary beat frequency (CBF) of nasal respiratory cells: ofloxacin as an antibiotic; Betadine and hydrogen peroxide (H2O2) as antiseptic; and amphotericin B, itraconazole, and clotrimazole as antifungal solutions. Differences are described between effects of each of these substances and we clarify whether ciliotoxic effects are dose dependent and if they can be reduced or eliminated by diluting the concentration of the applied solution. In a prospective study, nasal respiratory cells were harvested from healthy individuals and CBF was measured while cells were perfused with cell culture medium and the respective topical solution, using a Dvorak/Stotler exposure chamber. Controls were perfused with cell culture medium only. Video interference contrast microscopy was used to monitor CBF. A decrease of CBF occurred after application of all topical solutions used in this study. Except for clotrimazole, all solutions showed significantly stronger effects at high concentrations compared with diluted solutions. Our results indicate that topical application of antifungal, antibiotic, and antiseptic solutions may cause a marked impairment of mucociliary clearance. The strong dose dependence of these ciliotoxic effects indicates the need for a careful selection of the adequate concentration when using topical treatment in the nose. PMID- 11895192 TI - Posttransplant lymphoproliferative disorder of the nasopharynx. AB - Posttransplant lymphoproliferative disorder (PTLD) is a serious complication of organ transplantation. Although the precise etiology is unknown, the Epstein-Barr virus and immunosuppressive agents appear to he risk factors. The presentation PTLD is diverse, including many patients with symptoms of the head and neck, which may make diagnosis difficult. We present a patient who had undergone renal transplantation referred for recurrent sinusitis. She was found to have PTLD of the nasopharynx. Three cases of head and neck PTLD treated at our institution are described. Although PTLD is uncommon in the general community, the incidence has continued to increase as more patients undergo transplants and clinical presentations of PTLD should be familiar to the otolaryngologist. PMID- 11895194 TI - Acoustic rhinometry in the dog: a novel large animal model for studies of nasal congestion. AB - The aim of this project was to develop and pharmacologically characterize an experimental dog model of nasal congestion in which nasal patency is measured using acoustic rhinometry. Solubilized compound 48/80 (0.3-3.0%) was administered intranasally to thiopental anesthetized beagle dogs to elicit nasal congestion via localized mast cell degranulation. Compound 48/80-induced effects on parameters of nasal patency were studied in vehicle-treated animals, as well as in the same animals pretreated 2 hours earlier with oral d-pseudoephedrine or chlorpheniramine. Local mast cell degranulation caused a close-related decrease in nasal cavity volume and minimal cross-sectional area (Amin) together with a highly variable increase in nasal secretions. Maximal responses were seen at 90 120 minutes after 48/80 administration. Oral administration of the adrenergic agonist, d-pseudoephedrine (3.0 mg/kg), significantly antagonized all of the nasal effects of compound 48/80 (3.0%). In contrast, oral administration of the histamine H1 receptor antagonist chlorpheniramine (10 mg/kg) appeared to reduce the increased nasal secretions but was without effect on the compound 48/ 80 induced nasal congestion (i.e., volume and Amin). These results show the effectiveness of using acoustic rhinometry in this anesthetized dog model. The observations that compound 48/80-induced nasal congestion was prevented by d pseudoephedrine pretreatment, but not by chlorpheniramine, suggest that this noninvasive model system may provide an effective tool with which to study the actions of decongestant drugs in preclinical investigations. PMID- 11895193 TI - Cetirizine decreases interleukin-4, interleukin-5, and interferon-gamma gene expressions in nasal-associated lymphoid tissue of sensitized mice. AB - Although the action of cetirizine dihydrochloride (cetirizine), a potent histamine H1 receptor antagonist, has been well known, its effect on the cytokine profiles in the nasal immune inductive site has not been elucidated yet. We studied the effect of cetirizine on the cytokine profiles in the nasal-associated lymphoid tissue (NALT), which is a principal mucosal lymphoid tissue of the respiratory tract in rodents. Two different doses of cetirizine were given intraorally for 5 days before the nasal challenge of ovalbumin in sensitized mice. The sensitized group was given normal saline instead of cetirizine, and the nonsensitized group had no sensitization or medication. The cytokine gene expressions in the NALT taken from the mice were investigated with real-time quantitative reverse-transcription polymerase chain reaction. The effect of cetirizine on the allergic symptom score, histamine threshold, and the eosinophil count in the nasal septal mucosa were examined also. Compared with the normal mice, the sensitized mice showed significantly increased levels of interleukin (IL)-4 and IL-5 gene expression although the increase of interferon (INF)-gamma gene expression was not significant. In the cetirizine groups, the levels of expression of IL-4, IL-5, and INF-gamma in the NALT were significantly decreased compared with the sensitized group. The cetirizine groups also showed decreased allergic symptom score, histamine threshold, and eosinophil count in the nasal septal mucosa compared with the sensitized group. In conclusion, cetirizine reduced the levels of expression of IL-4, IL-5, and INF-gamma in the NALT of ovalbumin-sensitized mice. Cetirizine also reduced the acute allergic symptom, histamine sensitivity, and eosinophil count in the nasal septal mucosa. PMID- 11895195 TI - Endonasal endoscopic treatment of parasellar arachnoid cyst: report of a case. AB - A 40-year-old man presented with intractable headache of 5-year duration and a 1 month history of intermittent cerebrospinal fluid (CSF) rhinorrhea. Magnetic resonance imaging showed a cystic lesion with signal characteristics similar to that of CSF. The patient underwent endonasal endoscopic surgery of the sphenoid sinus and the fistula was reinforced with facia, muscle cartilage, and posterior septal flap while performing cystocisternostomy. The postoperative course was uneventfiul CSF leakage stopped, and headache improved. Postoperative imaging revealed total collapse of the cyst cavity. Based on our findings, endonasal endoscopic treatment of the sellar and parasellar arachnoid cysts, if presenting into the sphenoid sinus, could be an acceptable minimally invasive alternative to the conventional modalities. PMID- 11895196 TI - Histologic and immunologic observations of viral-induced rhinosinusitis in the mouse. AB - Viral upper respiratory infection is one of the most common diagnoses made in primary care offices. Although symptoms resolve within 1 week for many patients, a percentage develops rhinosinusitis, and many of these patients are treated with antibiotics. We have developed a model of viral rhinosinusitis using intranasal inoculation of reovirus into mice that were then killed on postinoculation days 2, 4, 7, 10, 14, or 21 and heads were embedded in paraffin for histological and immunohistochemical analyses. Reovirus-like immunoreactivity was noted in the septa and paranasal sinus mucosa in mice as early as day 2, with peak intensity seen on day 4, and scant staining seen on day 7. Complete absence of viral staining was seen by day 10, which corresponded with increased intracellular adhesion molecule 1 immunostaining in the nose. By day 10, a large mucosal influx of B cells was observed, with a moderate influx of macrophages and smaller influx of T cells. By day 14, there was a peak in the number of B cells with a corresponding, but less pronounced peak in T cells, while macrophages began to decline at this point. By day 21, the panel of immune markers returned to near normal levels. The results of this study suggest that the immune system continues to produce a response as long as 2 weeks after clearance of viral antigens. One proposed mechanism for this phenomenon is that local factors such as cytokines are released continually after infection, even in the absence of persistent viruses or bacteria. PMID- 11895197 TI - Analysis of the adult chronic rhinosinusitis working definition. AB - In 1997, the Task Force on Rhinosinusitis produced a working definition for adult chronic rhinosinusitis. The diagnostic criteria grew out of expert consensus opinion and were "intended to serve as a unifying starting point for further research." The validity of these criteria has not been tested yet. This article summarizes our experience with these diagnostic criteria. All patients presenting for evaluation of rhinosinusitis were routinely questioned as to the presence of a number of sinonasal symptoms, including those contained within the Task Force's working definition. The presence or absence of nasal purulence on anterior nasal examination was noted also. Patients who subsequently had surgery to address their chronic rhinosinusitis were felt to be the "gold standard" and comprised the study population. All patients included in the study had symptoms for > 12 weeks, evidence of rhinosinusitis that was discovered by computed tomography (CT), and inflammation found on analysis of pathology specimens. These patients' symptom questionnaires and nasal examinations were reviewed. Fifty-seven patients met the criteria for inclusion in the study. Nasal obstruction/blockage and facial congestion/fullness were the most common symptoms. The average numbers of major and minor symptoms were 3.2 and 2.8, respectively. Patients with polyps and with previous nasal or sinus surgery differed little from the overall population. Of the 57 patients in the study, 50 patients met the diagnostic criteria set forth by the Task Force for a sensitivity of 87.7%. Diagnosing chronic rhinosinusitis without radiological imaging or nasal endoscopy is difficult. The criteria set forth by the Task Force provide a relatively sensitive working definition of chronic rhinosinusitis. Further work is underway to assess the specificity and predictive value of these criteria. PMID- 11895198 TI - Arsenic trioxide in multiple myeloma: rationale and future directions. AB - Multiple myeloma remains an incurable malignancy with a median survival that does not exceed 3 years. At least one third of patients with multiple myeloma fail to respond to induction chemotherapy, and those who initially achieve remission eventually relapse and require additional therapy. Recent reports demonstrating the efficacy of arsenic trioxide in acute promyelocytic leukemia have prompted a revival in the clinical use of this compound. The achievement of clinical responses marked by molecular conversion of the malignant phenotype and remissions in patients who had failed to respond to multiple courses of conventional chemotherapy provided the impetus to explore its use in multiple myeloma. Properties that favor the use of arsenic trioxide are its ability to target selectively malignant cells for apoptosis through enhancementof reactive oxygen species, to induce differentiation, and to inhibit angiogenesis. Multiple events involved in the pathogenesis of multiple myeloma coincide with pathways targeted by arsenic trioxide, and early results have suggested that clinical responses and safety in patients are promising with advanced disease. PMID- 11895199 TI - Advances in allogeneic stem cell transplantation: directing graft-versus-leukemia at solid tumors. AB - Allogeneic stem cell transplantation was originally developed as a method to rescue hematopoietic function following high dose "myeloablative" therapy in the treatment of hematological malignancies. In the first two decades of its use, dose-intensive chemotherapy alone was credited with curing those patients who achieved sustained remission following this procedure. However, more recently investigators have come to recognize that antineoplastic effects mediated by immunocompetent donor T-cells transplanted with the stem cell allograft can be induced against hematological malignancies. Indeed, this graft-vs-leukemia (GVL) or graft-vs-tumor (GVT) effect is now felt to represent the principal modality required to sustain durable remissions of hematological malignancies following this approach. The powerful and potentially curative nature of the GVT effect in hematological cancers has recently lured oncologists into exploring the therapeutic potential of allogeneic stem cell transplantation as an investigational approach for treatment-refractory solid tumors. We review here the development and early clinical results of allogeneic stem cell transplantation as potential immunotherapy for solid tumors. PMID- 11895200 TI - Feasibility of thermal ablation of lytic vertebral metastases with radiofrequency current. PMID- 11895201 TI - Which results justify the means in prostate cancer treatment? PMID- 11895202 TI - An evidence-based analysis of the management of localized prostate cancer. AB - PURPOSE: Patients with localized prostate cancer and their doctors face complex trade-offs when deciding on treatment. In this study, we modify the "number needed to treat" method to compare radical prostatectomy with radiotherapy. METHODS: A MEDLINE search was performed to identify all studies of radical protatectomy or radiotherapy for prostate cancer. Number needed to treat formulas were modified to account for not only survival but also for complications and their utilities. RESULTS: The unadjusted number needed to treat value for overall survival was 6 favoring prostatectomy (six patients have to undergo prostatectomy to have one more 10-year survivor than if they had undergone radiotherapy). Radiotherapy patients were 4 years older than prostatectomy patients. Because overall survival is strongly linked to patients' age and overall health, the numbers needed to treat for disease-specific and distant metastasis-free survival were analyzed to minimize patient selection bias. The unadjusted number needed to treat values for disease-specific and distant metastasis-free survival were 14 and 18, respectively. When number needed to treat is adjusted for complications and utilities, its value for overall survival is 14, disease-specific survival is -25, and distant metastasis free survival is -22, these last two favoring radiotherapy. CONCLUSIONS: Utility-adjusted numbers needed to treat for prostatectomy and radiotherapy are greatly influenced by the likelihood of complications and the utility loss ascribed to them. When literature-reported values are used, radiotherapy is superior, but the differences in outcomes are small. With prostate-specific antigen screening and refined treatment methods, these values will change, and the modified number needed to treat can be used to evaluate, report and compare results. The consequences of treatment in terms of both survival and quality of life determine patient choice and physician recommendations. PMID- 11895203 TI - Vascular endothelial growth factor enhances endothelial cell survival and tumor radioresistance. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is an important mediator of endothelial cell proliferation and survival. The purpose of the present studies was to investigate the role of VEGF in the tumor response to ionizing radiation. METHODS: Two ras-transformed murine fibrosarcoma cell lines, VEGF+/+ and VEGF-/- were exposed to ionizing radiation (0, 1, 3, 5, 7 or 9 Gy) in vitro, and clonogenic survival was determined. VEGF+/+ and VEGF-/- xenografts were generated in athymic nude mice and then treated with ionizing radiation (ten 5-Gy fractions = 50 Gy). Mean fractional tumor volume was used to evaluate treatment efficacy. To determine whether VEGF enhances tumor radioresistance by targeting endothelial cells, we performed clonogenic survival assays with human umbilical vein endothelial cells. Surviving fractions were calculated after treatment with ionizing radiation (5 Gy) and recombinant hVEGF165 (0, 1, 10, and 100 ng/mL). To determine whether VEGF neutralization enhances tumor radiosensitivity, we employed anti-VEGF165 monoclonal antibody to treat human tumor xenografts. Tumors were exposed to ionizing radiation (four 5-Gy fractions = 20 Gy) and treated with anti-VEGF antibody (0, 5, and 25 microg/kg in four intraperitoneal doses). Mean fractional tumor volume was used to evaluate treatment efficacy. To elucidate the molecular mechanism contributing to the observed anti-VEGF/ionizing radiation interaction, we exposed human umbilical vein endothelial cells to ionizing radiation (5 Gy) in the presence of anti-VEGF antibody (1 microg/mL). Sodium dodecyl sulfate polyacrylamide gel electrophoresis of cell lysates was probed for mitogen-activated protein kinase (MAPK) and MAPK kinase (MEK1/MEK2). RESULTS: The in vitro radiosensitivities of the VEGF+/+ and VEGF-/- clones were equivalent (D0 = 146 vs 149). However, the VEGF+/+ xenografts were more resistant to the cytotoxic effects of ionizing radiation than the VEGF-/- xenografts. VEGF+/+ xenografts demonstrated a faster doubling time (4.5 vs 6.0 days) and a shorter growth delay (15 vs 23 days) than VEGF-/- xenografts. The surviving fraction of human umbilical vein endothelial cells after exposure to ionizing radiation was significantly enhanced in the presence of VEGF (6.4% vs 12.5%). Western blot analysis demonstrated that stimulation of MAPK and MEK1/MEK2 was abrogated after exposure to anti-VEGF antibody. DISCUSSION: These findings represent the first genetic evidence that factors other than inherent tumor cell radiosensitivity are important determinants of radiocurability. Antitumor strategies targeting VEGF and other endothelial cell survival mechanisms may be used to enhance the cytotoxic effects of radiotherapy. PMID- 11895204 TI - Dose/volume relationship of late rectal bleeding after external beam radiotherapy for localized prostate cancer: absolute or relative rectal volume? AB - PURPOSE: The purpose of this study was to analyze predictors of late rectal bleeding after external-beam radiotherapy for localized prostate cancer, with a focus on the volume of rectum irradiated. MATERIALS AND METHODS: One hundred twenty-eight patients were treated with external-beam radiotherapy at the Cleveland Clinic Foundation between January 1998 and June 1999. Conformal radiotherapy (CRT) was used to deliver 78 Gy at 2 Gy per fraction in 76 cases, and short-course intensity-modulated radiotherapy (SCIM-RT) was used to deliver 70 Gy at 2.5 Gy per fraction in 52 cases. All contours were determined by one physician. The rectum was outlined from 1 cm above the target structures to 1 cm below the target structures. The entire volume of the rectum, along with the outer rectal wall, was included. All cases had detailed planning parameters that specifically determined the rectal volume receiving the prescription dose (VrPr), that is, 78 Gy for CRT and 70 Gy for SCIM-RT, and the percent of rectal volume receiving the prescription dose (%VrPr). The RTOG scales were used to evaluate late toxicity. The median follow-up was 24 months for all cases (range, 3-34 months), 21 months for SCIM-RT cases (range, 11-26 months), and 28 months for CRT cases (range, 3-34 months). RESULTS: To date, five patients have had grade 1 late rectal toxicity (one CRT case and four SCIM-RT cases), one patient had grade 2 late rectal toxicity (CRT), and three patients had grade 3 late rectal toxicity (all CRT cases). Because of the low number of events, the analysis was performed with all patients experiencing rectal bleeding grouped together. The actuarial rectal bleeding rates at 18 and 24 months were 6% and 8%, respectively. The actuarial rectal bleeding rates at 24 months were identical (8%) for both SCIM-RT and CRT. A multivariate analysis of the following parameters was performed to determine independent predictors of rectal bleeding: age (continuous variable), race (Caucasian vs African American), coverage of seminal vesicles (yes vs no), adjuvant androgen deprivation (yes vs no), technique (CRT vs SCIM-RT), Radiation Therapy Oncology Group acute rectal toxicity score (continuous variable), VrPr (continuous variable in cubic centimeters), and %VrPr (continuous variable). Only the VrPr (cubic centimeter) was an independent predictor of rectal bleeding; %VrPr was not. With different cut-off levels being used, a VrPr of 15 cm3 was significant on univariate analysis; the actuarial rectal bleeding rates at 24 months for patients with a VrPr < or = 15 cm3 versus a VrPr > 15 cm3 were 5% versus 22%, respectively. CONCLUSION> In our study sample, which included both conformal and intensity-modulated radiotherapy patients, the volume of rectum receiving the prescribed radiation dose (the equivalent of 78 Gy) was an independent predictor of late rectal bleeding. The percent of rectal volume receivingthe full dose was not. Using actual volume rather than percent volume also avoids the dependence on the extent of rectal volume contours. We recommend 15 cm3 as the cut-off of the rectal volume not to exceed the prescription dose. The rectal bleeding rate at 2 years for cases with < 15 cm3 receiving the full dose was only 5%. PMID- 11895206 TI - High-dose-rate remote afterloading intracavitary brachytherapy for the treatment of extrahepatic biliary duct carcinoma. AB - PURPOSE: The purpose of this study was to determine whether a dose response exists for extrahepatic bile duct carcinoma (EBDC) when treated with increasingly higher radiation doses delivered via a combination of external beam radiation (EBRT) and high dose rate intracavitary brachytherapy (HDRIB). To establish the best tolerated dose of HDRIB. METHODS AND MATERIALS: Eighteen patients with pathologically proven, locoregional but unresectable or incompletely resected EBDC were studied from 1991-1998 in this phase I/II trial. All patients received EBRT, delivered via megavoltage photons at standard fractionation schedules, for a total dose of 45 Gy. The HDRIB was delivered using the nucleotron HDR remote afterloading unit with a 10 Ci Ir192 source. Each treatment of HDRIB delivered 7 Gy at 1 cm depth. The first group of eight patients received one treatment of HDRIB (Group 1, total dose = 52 Gy). The second group of six patients received two weekly treatments (Group 2, total dose = 59 Gy). The last group of four patients received three weekly treatments of HDRIB (Group 3, total dose = 66 Gy). HDRIB was delivered once weekly concomitant with the EBRT. Acute adverse reactions were evaluated after for each group of patients before escalating to the next higher dose level of HDRIB. RESULTS: The median follow up time for all 18 patients was 15 months. The median survival for all 18 patients was 12.2 months (range 2 to 79.6 months). Overall two-year survival was 27.8%. Three patients (16.7%) had survival of more than 5 years. Dose response is suggested by the median survival of the three groups (9, 12.2, and 20.3 months for Group 1, 2, and 3, respectively), although this did not reach statistical significance. Complete or partial response (>50% reduction in tumor size) was seen in 25% of patients receiving total of 52 Gy compared to 80% of patients (5 patients in Group 2 and 3 patients in Group 3) receiving greater than 59 Gy (P = 0.05). No patients developed Grade 4 complications. One patient in Group 2 developed Grade 3 toxicity after second treatment of HDRIB. CONCLUSION: High dose rate brachytherapy of 21 Gy in three divided weekly treatments, plus 45 Gy of external beam radiation is well tolerated. A dose response is shown with significant increase of PR and CR rate for dose >59 Gy. This modality of treatment appears to be safe and effective for inoperable extrahepatic biliary duct carcinoma. PMID- 11895205 TI - I-125 versus Pd-103 for low-risk prostate cancer: morbidity outcomes from a prospective randomized multicenter trial. AB - PURPOSE: The purpose of this study was to test the hypothesis that the shorter half-life of Pd-103 versus I-125 results in a shorter duration of radiation related symptoms after prostate brachytherapy. METHODS: As of February 2000, 110 of a planned total of 380 patients with 1997 American Joint Commission clinical stage T1c-T2a prostatic carcinoma (Gleason grade 2-6, prostate-specific antigen, 4-10 ng/mL) had been randomly assigned to implantation with I-125 (144 Gy, TG-43) or Pd-103 (125 Gy, NIST-99). Isotope implantation was performed by standard techniques, using a modified peripheral loading pattern. Treatment-related morbidity was monitored by mailed questionnaires, using standard American Urologic Association (AUA) and Radiation Therapy Oncology Group criteria at 1, 3, 6, 12, and 24 months. Use of alpha-blockers to relieve obstructive symptoms was not controlled for but was noted at each follow-up point. All patients reported here have a minimum 1-year follow-up. Randomization was carried out at a central enrollment office where eligibility criteria were confirmed and the patient assigned by computerized random number generator to one of the two treatment arms. Patients were assigned to 95 blocks of four. Most statistical comparisons shown here are by Student's unpaired t-test at specific follow-up times, as indicated in the figure legends. Additionally, considering the patients' scores change overtime, repeated measures were incorporated in a mixed model assuming an unstructured covariance matrix. RESULTS: Patients in each arm were well matched by preimplant prostate volume, AUA score, and age. The AUA scores peaked at the 1 month point for both isotopes and then gradually declined. The difference was greatest at 6 months, when I-125 patients had a mean AUA score of 16 (+/- 8), compared with 11 (+/- 10) for the Pd-103 patients. By 12 months, mean AUA scores for the Pd-103 patients had decreased to 12 (+/- 9), compared with 13 (+/- 8) for the I-125 patients. At 6 months after implantation, 41% of Pd-103 patients were still taking alpha-blockers, versus 44% of I-125 patients. The differences between isotopes were more marked in patients with a low pretreatment AUA score or smaller preimplant transrectal ultrasonography volume. Results of the mixed model, incorporating repeated measures for each patient, showed that the effect of isotope choice on AUA score depended on time. This effect was further dependent on baseline AUA score, but not on transrectal ultrasonography volume or on age. Urinary and rectal morbidity was generally low, typically grade 1 or 2. There was a trend to greater morbidity with I-125 than with Pd-103, most markedly at the 6-month time point. DISCUSSION: Patients treated with Pd-103 recovered from their radiation-induced prostatitis sooner than I-125 patients. It appears that patients with minimal pretreatment urinary obstructive symptoms are the most likely to experience implant-related exacerbations of their symptoms and are the most likely to benefit from the more rapid half-life of Pd-103 rather than I-125. PMID- 11895207 TI - Perineural invasion is not predictive of biochemical outcome following prostate brachytherapy. PMID- 11895208 TI - The endocrinology of menstruation--a role for the immune system. AB - The human endometrium displays characteristic features, both structural and functional, across the menstrual cycle. It is the sex steroid hormones, oestrogen and progesterone, that drive the endometrium through the different phases of the cycle. Oestrogen and progesterone act sequentially to regulate cellular concentrations of their respective receptors, this interaction initiates gene transcription. Thereafter a cascade of local events prepares the endometrium for implantation, but in the absence of pregnancy, progesterone withdrawal leads to menstruation and cyclic repair. Withdrawal of progesterone from an oestrogen progesterone primed endometrium is the initiating event for the cascade of molecular and cellular interactions that result in menstruation. Progesterone withdrawal first affects cells with progesterone receptors. Early events in the menstrual process are vasoconstriction and cytokine up-regulation. The activation of lytic mechanisms is a later event and involves cells that may lack progesterone receptors, for example, uterine leucocytes and epithelial cells. Hence progesterone withdrawal results in a local increase of inflammatory mediators and the enzymes responsible for tissue breakdown. The total complex of local factors implicated in normal menstrual and aberrant menstrual bleeding are yet to be fully defined. PMID- 11895209 TI - Graves' disease, thyroid nodules and thyroid cancer. PMID- 11895210 TI - Are we in danger from an epidemic of Cushing's syndrome? Nature knows its course. PMID- 11895212 TI - Colorectal neoplasia in acromegaly. PMID- 11895211 TI - Risk of colorectal neoplasia in acromegaly: an independent view. PMID- 11895213 TI - Screening colonoscopy for acromegaly in perspective. PMID- 11895214 TI - Prevalence and incidence of hypopituitarism in an adult Caucasian population in northwestern Spain. AB - OBJECTIVE: To determine the prevalence and incidence of hypopituitarism in the general population. POPULATION: The study population comprised an average population sample of 146,000 adult inhabitants in South Galicia (northwestern Spain). The Medical Register of the General Hospital of Vigo ensured virtually complete case ascertainment for diagnosed hypopituitarism in this sample population. Only patients residing in the study area were included. The diagnosis of hypopituitarism was based on baseline and hormonal dynamic tests. DESIGN: The study comprised two cross-sectional surveys, the first from January to December 1992 and the second from January to December 1999, together with a longitudinal survey performed between January 1993 and December 1999. MAIN RESULTS: In the first survey the prevalence of hypopituitarism was 29/100,000 (CI, 19.88-37.72), without sex differences. In the second survey, the prevalence observed was higher than in the first, 45.5/100,000 (CI, 34.92-56.08). In the second survey, which included almost all cases registered in the first study, the cause of hypopituitarism was a pituitary tumour in 61%, a non-pituitary tumour in 9% and a non-tumour cause in 30%. Around 50% of patients had 3-5 pituitary hormonal deficiencies, with LH/FSH being the most prevalent. Patients with tumour-induced hypopituitarism showed a tendency to suffer GH deficiency more frequently than those due to non-tumour causes. In the longitudinal study with a population of 1,020,764 people-years of observation, the average annual incidence rate of hypopituitarism was 4.21 cases/100,000 (CI, 2.95-5.47), with this incidence being similar for both sexes. The annual incidence of hypopituitarism remained stable during the study period. CONCLUSION: We present for the first time data on the prevalence and incidence of hypopituitarism in the general adult population. These patients showed a tendency to suffer LH/FSH deficiency as the most prevalent hormone deficit. Furthermore, patients with hypopituitarism due to a tumour or its treatment showed a greater tendency to suffer GH deficiency than those with a non-tumour cause. These data may be useful for producing a rational programme for patients suffering from this condition and also for comparison with future data in our country and elsewhere in the world. PMID- 11895215 TI - Low individualized growth hormone (GH) dose increased renal and cardiac growth in young adults with childhood onset GH deficiency. AB - OBJECTIVE: In childhood onset GH deficiency (GHD) a reduction in left ventricular mass (LV-mass) and impairment of systolic function as well an impairment in glomerular filtration rate (GFR) has been shown. The aim of the present study was to assess if a low GH dose resulted in an improvement in morphological and functional parameters of these organs. DESIGN AND PATIENTS: Eleven patients with childhood onset GHD were investigated before and after 10 months of GH treatment at a dose of 1.5 IU/day (range 1-2), corresponding to 0.02 IU/kg/day or 7 microg/ kg/day. The GH dose resulted in a serum IGF-I level in the normal range in all but one patient. MEASUREMENTS: Doppler echocardiography of the heart and ultrasound examination of the kidneys was performed. Glomerular filtration rate (GFR) was estimated with iohexol clearance and urinary proteinuria was measured with 24-h urinary samples collected for analyses of albumin, alpha-1 microglobulin, IgG and albumin/creatinine clearance ratio. Body composition was measured by bioelectric impedance analysis. RESULTS: L V-mass index increased significantly after GH treatment (P = 0.04), and there was a clear trend for a positive correlation between the increase in serum IGF-I and the increase in LV mass index, although it did not reach significance (r= 0.57, P = 0.07). GH treatment did not increase cardiac fractional shortening. Kidney length increased significantly (P = 0.02) with an average increase of 1 cm (range - 0.5-1.5 cm). No significant changes in median GFR or serum creatinine were recorded. Three patients with subnormal GFR before GH treatment normalized after 10 months of treatment. Urine analysis showed no abnormalities before or after GH treatment. A significant decrease in percentage fat mass was recorded (P = 0.03). CONCLUSION: A low individualized GH dose to adults with childhood onset GHD resulted in an increase in LV-mass index and kidney length. Re-establishing GH treatment with a low dose in this patient group can lead to a further somatic maturation of these organs, probably not accomplished previously. PMID- 11895216 TI - Effect of long-term administration of recombinant human growth hormone (rhGH) on plasma erythropoietin (EPO) and haemoglobin levels in anaemic patients with adult GH deficiency. AB - OBJECTIVE: We investigated the effect of recombinant human GH (rhGH) on erythropoietin (EPO) and haemoglobin (Hb) concentrations in anaemic patients with adult GH deficiency. PATIENTS AND DESIGN: rhGH was administrated in 8 patients with adult GH deficiency, three males and five females, aged from 24 to 69 years, mean (+/- SD) of 48.8 +/- 16.4 years, for 1 year by means of continuous subcutaneous infusion (CSI) at a flow rate of 0.036 U/kg/day using a portable syringe pump. Blood samples were obtained in the morning after an overnight fast every week for 1 month, followed by each month before and after the start of rhGH administration. RESULTS: Mean (+/- SE) plasma GH levels increased from 0.24 +/- 0.09 microg/l to 2.32 +/- 0.23 microg/l 1 week after the start of rhGH administration to maintain a steady state. Plasma IGF-I levels increased from 70.1 +/- 13.8 microg/l to 282.8 +/- 70.6 microg/l 1 week after the start of rhGH administration to maintain the steady state. Plasma EPO levels increased from 25.9 +/- 2.6 IU/l to 37.6 +/- 4.2 IU/l and 34.3 +/- 3.6 IU/l at 1 week and 2 weeks after the start of rhGH administration, respectively, and then decreased gradually to 14-9 +/- 2.1 IU/l at 10 months after the start of rhGH administration. Reticulocyte counts increased from 0.88 +/- 0.06% to 1.49 +/- 0.21% at 1 week. Hb concentrations increased from 103 +/- 5 g/l to 106 +/- 5 g/l at 2 weeks after the start of rhGH administration, and then increased gradually to reach the normal range. CONCLUSIONS: We conclude that EPO secretion was stimulated in the initial 2 weeks after the start of CSI of rhGH in anaemic patients with adult GH deficiency. Increased Hb concentrations after long-term administration of rhGH might be explained by direct stimulatory effects of rhGH and IGF-I on erythroid cells, which was accompanied by suppressed EPO secretion, in combination with a more generalized indirect impact of rhGH on physical activety. These findings suggest a beneficial effect of rhGH replacement in anaemic patients with adult GH deficiency. PMID- 11895217 TI - Effect of digoxin on the somatotroph responsiveness to growth hormone-releasing hormone (GHRH) alone or combined with arginine in normal young volunteers. AB - BACKGROUND: The activity of the GH/IGF-I axis, known to play a major role in myocardial structure and function, has been reported to be altered in patients with chronic heart failure. AIM AND DESIGN OF THE STUDY: In order to evaluate the possibility that clinically used cardioactive drugs may exert neuroendocrine influences on somatotroph secretion, we studied the effects of pretreatment with enalapril (20 mg/day orally for 3 days), furosemide (20 mg i.v. as a bolus at -5 minutes) or digoxin (0.25 mg orally 4x/day for 3 days) on the GH response to growth hormone-releasing hormone (GHRH) (1.0 microg/ kg i.v. as a bolus at 0 minutes) in 12 healthy male adults (age [mean +/- SEM] 30.2 +/- 1.4 years; BMI 22.7 +/- 0.7 kg/ m2). In a subgroup of 8 subjects the same study was performed testing the GH response to GHRH + arginine (ARG; 0.5 g/kg i.v. from 0 to + 30 minutes). RESULTS The GH response to GHRH (1,304.1 +/- 248-5 microg/l/h) was not modified by enalapril (1,368.7 +/- 171.2 microg/l/h) or by furosemide (1,269.3 +/ 185.2 microg/l/h) but was significantly blunted by digoxin (613.6 +/- 73.2 microg/l/h, P < 0.05). On the other hand digoxin, enalapril and furosemide did not modify the GH response to GHRH +ARG. CONCLUSIONS: Digoxin, but not enalapril or furosemide, inhibits the GH response to GHRH in normal subjects. The blunting effect of digoxin on the GHRH-induced GH response is counteracted by arginine. These findings show that digoxin possesses an inhibitory effect on somatotroph secretion that may be mediated at the hypothalamic level. PMID- 11895218 TI - The contribution of IGF-I to skeletal integrity in postmenopausal women. AB - OBJECTIVES: The pathogenic role of the decline in serum concentrations of IGF-I in postmenopausal osteoporosis is not fully elucidated. We investigated the associations among IGF-I, bone mineral density (BMD), ultrasound parameters and prevalence of vertebral fractures in postmenopausal women. DESIGN: A cross sectional study. PATIENTS: One hundred and fifty-four ambulatory postmenopausal women (61 +/- 7 years) referred for osteoporosis screening. MEASUREMENTS: IGF-I was measured by radioimmunoassay and BMD using dual-energy X-ray absorptiometry. Broadband ultrasound attenuation (BUA) and speed of sound (SOS) at calcaneus were measured by a quantitative ultrasound system. RESULTS: IGF-I was significantly lower in osteoporotic subjects and correlated positively with BMD, BUA and SOS. After adjusting for age, years since menopause and body mass index, IGF-I accounted for 8.5% of the variance at lumbar spine BMD, 4.6% at femoral neck and 7.1% at calcaneal BUA. BUA was associated with IGF-I independently of BMD. IGF-I was lower in women with vertebral fractures (91 +/- 39 microg/l vs. 114 +/- 44 microg/l; P = 0.003). The osteoporosis densitometric criteria (t-score < or = 2.5 SD) was the most strongly independent associated variable with prevalent vertebral fractures [odds ratio (OR): 3.3 (1.4-7.6)], followed by IGF-I levels below 75th percentile [OR: 3 (1-8.8)]. CONCLUSIONS: Our study shows that IGF-I is strongly associated with bone mineral density and reflects aspects of bone quality. The contribution of IGF-I to skeletal integrity in postmenopausal women is clinically relevant. PMID- 11895219 TI - Obesity, rather than menstrual cycle pattern or follicle cohort size, determines hyperinsulinaemia, dyslipidaemia and hypertension in ageing women with polycystic ovary syndrome. AB - OBJECTIVE: The aim of this study was to investigate if ageing women with polycystic ovary syndrome (PCOS) who gained regular menstrual cycles differed from women who continued to menstruate irregularly with regard to risk factors for developing diabetes mellitus and atherosclerosis. DESIGN AND PATIENTS: In the original study of a population of 346 PCOS patients, defined in the past as having oligo- or amenorrhoea and elevated LH concentrations, we had sent out a questionnaire to investigate changes in the pattern of their menstrual cycles while ageing. From this cohort of patients, a significantly older group of 53 women (mean age: 41.3 years, range: 33.3-49.4) who were not using oral contraceptives or other hormones visited the outpatient clinic. These women did not differ from the non-participating group in BMI, ethnic origin, the proportion with regular menstrual cycles by age group, parity or the use of clomiphene citrate or gonadotrophins in the past. MEASUREMENTS: A physical examination and a transvaginal ultrasound were performed. The size of the follicle cohort was determined by counting the number of small follicles in the ovaries. Thirty-four women were also willing to give two fasting blood samples for measuring their glucose, insulin and lipid status. RESULTS: Forty-one of the 53 (77.4%) women had a regular menstrual cycle (shorter than 6 weeks) and 12 (22.6%) had an irregular cycle (longer than 6 weeks). The body mass index (BMI), waist: hip ratio (WHR), systolic blood pressure (SBP), diastolic blood pressure (DBP) and prevalence of diabetes (1-9%) and hypertension (11.3%) did not differ between the two menstrual cycle groups. Also, the fasting glucose, insulin, glucose/insulin ratio, total cholesterol, HDL-c, and LDL-c concentrations did not show any significant difference between the two groups. Instead, these parameters all were significantly higher in women with a BMI > 27 kg/M2 compared to women with a BMI < or = 27 kg/m2. Regularly menstruating PCOS women were older (P < 0.01), showed less follicles in their ovaries (n = 48, P < 0.01) and had lower androgens (n = 34, P < 0.05) than the irregularly menstruating women. Logistic regression analysis showed a second significant influence, after age, of the BMI on the menstrual cycle pattern (age, P < 0.01; BMI, P < 0.05). If age was excluded from the analysis, only the follicle count significantly predicted the menstrual cycle pattern (P < 0.02). CONCLUSIONS: We conclude that hyperinsulinaemia, dyslipidaemia and hypertension in our population of ageing women with polycystic ovary syndrome are not related to the menstrual cycle pattern but rather to obesity. Age and the size of the follicle cohort are the main factors determining the menstrual cycle pattern in ageing women with polycystic ovary syndrome, although an association with the BMI was also found. PMID- 11895220 TI - Normal VLDL metabolism despite altered lipoprotein composition in type 1 diabetes mellitus. AB - OBJECTIVES: Patients with type 1 diabetes are at increased risk of cardiovascular disease, which may be related to abnormal lipid metabolism. Secretion and clearance of VLDL apolipoprotein B100 (apoB) are important determinants of plasma lipid concentrations and are known to be influenced by hormones, including insulin and growth hormone. PATIENTS: This study examined overnight VLDL apoB metabolism and VLDL composition in six lean patients with type 1 diabetes during euglycaemia (controlled by a varying insulin infusion) and in six age-, sex- and BMI-matched control subjects. METHODS: VLDL apoB kinetics were determined using a primed constant 1-13C leucine infusion, and VLDL apoB enrichment was measured by gas-chromatography mass-spectrometry. Fasting lipid profile, IGF-I, IGFBP-3, overnight GH profiles and free insulin concentrations were also assessed. RESULTS: Fasting concentrations of triglycerides (TG), total cholesterol (TC), HDL-cholesterol (HDL-C) and LDL-cholesterol (LDL-C) were similar in both groups. The VLDL apoB secretion and metabolic clearance rates were not significantly different between the two groups, but the VLDL-TGNLDL apoB and the VLDL-CNLDL apoB ratios were significantly increased in those with diabetes (P < 0.02 and P < 0.03, respectively). Total IGF-I concentrations were similar between the two groups; however, the GH area under the curve and free insulin concentrations were increased in patients with type 1 diabetes (GH: diabetes: 94.8 +/- 15.1 vs. controls: 45.6 +/- 10-6, mU/L/h, P < 0.04; free insulin: diabetes: 78.4 +/- 5.0 vs. controls: 28.3 +/- 3.26, pmol/l, P < 0.001). IGFBP-3 concentrations were lower in diabetic patients (diabetes: 2,454.2 +/- 68.7 vs. controls: 3,219.4 +/- 76.4, ng/ml, P < 0.001). In the control group overnight GH secretion correlated negatively with fasting TC (P < 0.01) and LDL-C (P < 0.03) concentrations, whereas free insulin concentrations correlated positively with fasting TG concentrations (P < 0.009). No significant correlations were found in the patients with diabetes. CONCLUSION: This study suggests that in euglycaemic conditions patients with type 1 diabetes mellitus have normal VLDL apoB kinetics but altered VLDL composition. The altered VLDL composition may be associated with accelerated atherogenesis. We speculate that the disrupted hormonal balance and, in particular, the increased GH secretion might be responsible for the compositional changes of VLDL particles in type 1 diabetes mellitus. PMID- 11895221 TI - An ultrasensitive assay revealed age-related changes in serum oestradiol at low concentrations in both sexes from infancy to puberty. AB - OBJECTIVE: Intensive studies of oestrogen receptors have suggested extragonadal functions of oestrogen. However, the in vivo extragonadal functions of oestradiol remain unclear because of the lack of an adequate assay system at low concentrations. In this study, we assessed the usefulness of a new ultra sensitive assay for children. METHODS: Serum oestradiol was measured with an ultrasensitive assay (assayable concentration: 5-1,835 pmol/l: ESTR-US-CT, CIS biointernational, France). Intra- and interassay coefficients of variation at low concentrations (< 36.7 pmol/l) were 8.2 +/- 6.8 (0.1-31.2)% and 8.3 +/- 3.7 (7.5 12.9)%, respectively. SUBJECTS: Sera from 88 healthy children (55 males and 33 females; 1 month to 16 years old) and 31 patients who underwent gonadal suppression therapy were analysed. RESULTS: Age-related changes were observed in both sexes. Serum oestradiol concentrations in childhood decreased slightly compared to those in infancy, then increased at puberty. Most prepubertal children showed oestradiol concentrations lower than 36.7 pmol/l. A study on patients who underwent gonadal suppression therapy revealed oestradiol changes within low concentrations, depending on the stage of the therapy. CONCLUSIONS: The new assay was considered precise enough for the assessment of oestradiol secretion at low concentrations in childhood. Age-related changes in serum oestradiol suggested gonadal activity in the prepubertal period. This assay could be a powerful tool for investigating novel oestradiol functions in vivo. PMID- 11895222 TI - Dehydroepiandrosterone sulphate is increased and dehydroepiandrosterone-response to corticotrophin-releasing hormone is decreased in the hyperthyroid state compared with the euthyroid state. AB - OBJECTIVE: Dehydroepiandrosterone (DHEA) and DHEA-sulphate (S) have been suggested to play protective roles in many pathological states, some of which are observed in hyperthyroidism. If DHEA and DHEA-S levels change in hyperthyroidism, they might participate as a possible causative link with such pathophysiological changes in hyperthyroidism. However, the CRH-ACTH-DHEA system in hyperthyroidism has not been clearly defined. We examined plasma levels of DHEA and DHEA-S together with ACTH and cortisol in both hyperthyroid (Hyper) and euthyroid states (Eu). METHODS: Eighteen patients (5 men and 13 women, aged 46.9 +/- 2.8 years) with Graves' disease were studied before treatment and again in the euthyroid state following treatment with methimazole. A 100 microg hCRH stimulation test and a low-dose (0.5 microg) 1-24 ACTH stimulation test were performed on separate days. Basal levels and A area under the response curve (AUC) were compared between Hyper and Eu. RESULTS: DHEA-S was higher in Hyper than in Eu. However, basal DHEA did not differ between Hyper and Eu. The ratio of DHEA to DHEA-S was lower in Hyper than in Eu. AAUC of DHEA during a CRH test was lower in Hyper than in Eu. However, AAUC of DHEA during an ACTH test was similar in both Hyper and Eu. Basal ACTH was higher in Hyper than in Eu. In both CRH and ACTH tests, AAUC of cortisol response was lower in Hyper than in Eu, although the basal cortisol level was not different. CONCLUSION: The balance of the conversion between DHEA-S and DHEA in the hyperthyroid state favoured DHEA-S. Similar to cortisol, the DHEA response in the CRH test in hyperthyroidism seemed to be insufficiently compensated for by increased ACTH, although the DHEA response to low-dose ACTH was similar in the hyperthyroid and euthyroid states. Increased DHEA-S might play some role in the pathological states in many organs in hyperthyroidism. PMID- 11895223 TI - HLA-DRB3*0101 is associated with Graves' disease in Jamaicans. AB - OBJECTIVES: Graves' disease is associated with different human leucocyte antigen (HLA) genes in different populations. This studywasdesigned to examinethe HLA class II associations with Graves' disease in Jamaicans. PATIENTS: One hundred and six Jamaicans with Graves' disease and 104 controls. DESIGN: Oligotyping for HLA-DRB1, DRB3, DQA1 and DQB1 alleles was performed using the polymerase chain reaction sequence specific oligonucleotide probe (PCR-SSOP) technique. RESULTS The frequency of HLA-DRB3 *0101 was increased significantly in the patients compared to controls (38.7% vs. 19.2%; RR = 2.72; Pc < 0.015). The protective alleles for Graves' disease were DRB1 *0901 (0.9% vs. 20.2%; RR = 0.04; Pc < 0.001), DRB1*1001 (0.0% vs. 11%; RR = 0.0%; Pc < 0.01) and DRB4 *0101 (0.0% vs. 12.5%; RR = 0.0; Pc < 0.05). A high female to male ratio of Graves' disease, 25 :1, was observed. Other associated autoimmune diseases were rare and no significant HLA class II associations were found with clinical markers of disease. CONCLUSIONS: Jamaican patients with Graves' disease share the DRB3 *0101 susceptible allele and the DRB4 *01 protective allele but not the susceptible haplotype DRB1 *0301, DRB3*0101, DQA1*0501 with Caucasians. PMID- 11895224 TI - Postpartum thyroid dysfunction and postpartum depression: are they two linked disorders? AB - OBJECTIVE: Postpartum has been considered as a period of risk for developing postpartum depression (PD) by some but not all authors, and this PD has been linked with postpartum thyroid dysfunction (PPTD). The major aim of this study was to evaluate the relation between the presence of PPTD and PD. DESIGN AND PATIENTS: Six hundred and forty-one healthy Caucasian women recruited between their 36th week of pregnancy and fourth day postpartum underwent clinical and laboratory evaluation and were checked again at 1 (n = 605), 3 (n = 552), 6 (n = 574), 9 (n = 431), and 12 (n = 444) months postpartum. MEASUREMENTS: At baseline and at each clinical evaluation, Beck Depression Inventory (BDI) was administered to screen PD. The definitive diagnoses of PD was performed by a psychiatrist according to the DSM-III-R criteria. At each visit, we determined serum free T4 and TSH concentrations. Thyroperoxidase and thyroglobulin antibodies were determined only in patients with abnormal hormone concentrations. Postpartum thyroiditis (PPT) was considered to be present in women with overt or subclinical transient hyperthyroidism between 1 and 3 months postpartum and/or overt or subclinical hypothyroidism between 3 and 6 months postpartum. RESULTS: Fifty-six women developed postpartum thyroid dysfunction (PPTD), corresponding to an incidence rate of 11%: 45 with PPT [incidence rate 7.8%; confidence interval (CI) 5.6-10%], eight with Graves' disease (incidence rate 1.5%; CI 0.5-2.5%) and three with nonpalpable toxic thyroid adenoma (incidence rate 0.5%; CI 0-1.5%). Five hundred and eighty of the evaluated women (incidence rate 90.5%; CI 95% 88.2 92.8) presented BDI scores below 21 and therefore the PD diagnoses was excluded. In 50 cases (incidence rate 7.8%; Cl 95% 5.7-9.8), we detected a BDI score over 21 in some evaluations, but the PD diagnosis was not confirmed. Another 11 (incidence rate 1.7%; CI 95% 0.7-2.7) were diagnosed as having PD and required psychiatric treatment. None of the PPTD was diagnosed as having PD. The BDI scores frequency over 21 was similar between healthy women and those with PPTD. Patients with a previous history of depression developed PD more often (P < 0.0001). One hundred and ninety women breast fed their babies for more than 2 months, without observing a higher PD rate or BDI scores over 21 (P = 0.5). CONCLUSIONS: We found a general PD incidence rate of 1.7% in our group of patients. This figure is not higher in women with hormone abnormalities caused by PPTD. Women with a past history of depression present a higher risk of PD while those who breast fed did not have an increased risk. PMID- 11895225 TI - Chronic hypocalcaemia due to selective skeletal resistance to parathyroid hormone. AB - A 71-year-old man was referred for evaluation of asymptomatic hypocalcaemia dating back at least 20 years. There were no somatic abnormalities and Chvostek and Trousseau signs were negative. Serum total calcium varied from 1.88 to 2.03 mmol/l, albumin 37-44 g/l, phosphate 0.54-1.12 mmol/l and ionized calcium 1-1.13 mmol/l. Serum intact PTH levels were 69 and 55 ng/l (10-65), 25-OHD was 40 nmol/l (2.25-107.5) and 1,25-(OH)2D was 54.6 nmol/l (39-156). Serum and urine magnesium and creatinine clearance were normal. Twenty-four-hour urine calcium was 2.15 mmol and calcium/creatinine ratio 0.07. TM phosphate (maximal rate of tubular reabsorption of phosphate in mmol/l glomerular filtrate (GF)) was 0.84 mmol/l GF (0.80-1.34). Bone formation and resorption markers were normal. Bone mineral densities measured by dual-energy X-ray absorptiometry (DEXA) were within normal limits at the hip, forearm and lumbar spine. Infusion of 200 units of synthetic 1 34 PTH was associated with a rise in urinary cyclic AMP from 43 mmol/l GF to 344 mmol/l GF and TM phosphate fell from 0.93 to 0.76 mmol/l GF; 1-34 PTH infusions of 300 units twice daily for 5 days were associated with an increase in serum 1,25-(OH)2D from 80.6 to 114.4 pmol/l but no increase in serum calcium. This is a most unusual case of chronic hypocalcaemia similar to that reported by Frame et al. resulting from isolated skeletal resistance to PTH that is not related to renal insufficiency, osteomalacia or a magnesium-deficient state. These two cases appear to represent a new variant of pseudohypoparathyroidism ?type III. PMID- 11895226 TI - Management of Nelson's syndrome: observations in fifteen patients. PMID- 11895227 TI - Peak cortisol response to insulin tolerance test: how reproducible is it really? PMID- 11895229 TI - Evaluating the effects of research in medical education. PMID- 11895228 TI - Hypothalamo-pituitary-adrenal axis testing: nothing is sacred and caution in interpretation is needed. PMID- 11895230 TI - Modernising regulation in the health professions. PMID- 11895231 TI - The daily grind--use of log books and portfolios for documenting undergraduate activities. PMID- 11895232 TI - Qualitative methods and guideline implementation--a heady mixture? PMID- 11895233 TI - What do students actually do on an internal medicine clerkship? A log diary study. AB - BACKGROUND: There are limited data on the amount of time students spend on teaching and learning while on internal medicine clerkships, and existing data suggest a wide international variation. Community-based teaching of internal medicine is now widespread; but its strengths and weaknesses compared to traditional hospital based teaching are still unclear. AIM: To determine the proportion of time students spend on different activities on an internal medicine clerkship, and to determine whether this differs in general practice and in hospital. In addition we aimed to determine students' views on the educational value and enjoyment of various activities. METHODS: Prospective completion of log diaries recording student activities. Each student was asked to complete the diary for two separate weeks of their internal medicine clerkship: one week of general practice-based teaching and one week of hospital-based teaching. RESULTS: The response rate was 68% (88/130). Students spent approximately 5.5 h per day on teaching and learning activities in both environments, with more time (50 min vs. 30 min, P = 0.007) on unsupervised interaction with patients in hospital than in general practice, and more time (53 min vs. 21 min, P < 0.001) undergoingassessment in general practice than in hospital. Standard deviations were wide, demonstrating the heterogeneous nature of the data. Students perceived supervised interaction with patients and teaching by doctors as the most educational activities in both environments, but found it even more educationally valuable and enjoyable in general practice than in hospital (mean score for educational value: 4.27 in general practice, 3.88 in hospital, P = 0.048; mean score for enjoyment 4.13 in general practice, 3.66 in hospital, P = 0.03). CONCLUSIONS: Students greatly value interactions with patients, perceiving these as both educational and enjoyable. Curriculum planners must continue to place patient-based learning at the centre of undergraduate medical education. The heterogeneity of the data suggests that individual students have very different experiences, despite apparently similar timetables. PMID- 11895234 TI - 'Holding it lightly': the co-operative inquiry group: a method for developing educational materials. AB - SETTING: The purpose of this research was to adapt the World Health Organization's educational programme Mental Disorders in Primary Care for South African general practitioners. AIM: This paper describes how to organise and facilitate a co-operative inquiry group as a form of participatory action research aimed at developing or adapting educational materials. Specific quality criteria for this type of action research are defined. The experience of our own co-operative inquiry and the lessons learnt are discussed. CONCLUSION: In the field of medical education participatory action research methodology is relatively new. This article shows how the co-operative inquiry group can be used effectively to develop educational materials. It is intended to encourage and support others in using similar methods of action research in their own settings. PMID- 11895235 TI - Sheffield Assessment Instrument for Letters (SAIL): performance assessment using outpatient letters. AB - CONTEXT: Well-designed assessments of performance are urgently required for training doctors, and to provide indicators of the quality of practice. OBJECTIVES: To design an assessment process that uses routine outpatient letters, and to evaluate its validity, feasibility, reliability (reproducibility and discrimination) and potential educational impact. PARTICIPANTS: All 26 paediatric registrars in North Trent attending annual assessment panel in 1999 participated. STUDY DESIGN: An assessment instrument (SAIL) was developed from a consensus framework for good practice in written communication. It comprises an 18-point checklist and a global rating scale. Three judges applied the instrument to 260 letters from the routine clinical practice of the 26 participants. Results We achieved consensus on good practice in written communication. This was in keeping with the published literature. All participants completed the assessment. Scoring took 3-6 min per judge per letter. The reliability coefficient in this test situation is 0.72. Modelling predicts that a coefficient of 0.8 (the threshold for high-stakes judgements about performance) can be achieved with more judges or letters. The assessment results are well suited to formative feedback. CONCLUSIONS: SAIL uses letters as a face valid indicator of written communication performance. The instrument is feasible to use, and produces reliable results when applied to paediatric registrars to inform the annual Record of In-Training Assessment (RITA). Feedback from the assessment should help doctors to improve their written communication. Its use may extend to other specialities and settings including revalidation. PMID- 11895236 TI - Portfolios as a learning tool in obstetrics and gynaecology undergraduate training. AB - CONTEXT: We developed a structured portfolio for medical students to use during their obstetrics and gynaecology undergraduate training. The main objective was to support the learning process of the students. We also wanted feedback information to enhance teaching. METHODS: The study population consisted of 91 medical students who completed the portfolio during their training course. The portfolio consisted of a 28-page A5-size booklet. The students entered all the clinical procedures they had performed and all the deliveries they had attended. After each group session, they answered questions about what they had learned and evaluated the performance of the teacher. They also indicated their general evaluation of the course and the portfolio itself. The teachers listed the 13 most important skills to be learned during the course. The students were asked to evaluate their own development on a scale of 0-5 before and after the course. A content analysis was performed on all the texts the students produced, and all quantitative variables were coded. RESULTS: The amount of text written in the portfolio correlated (P < 0.001, F-value 4.2) with success in the final exam. In addition to acting as a logbook, use of the portfolio enhanced the learning process during the course. Students' attitudes towards the portfolio were mainly positive. Students appreciated the departmental interest in their learning process. CONCLUSION: Portfolios support the personal and professional development of medical students. A portfolio clarifies the learning goals and helps students to monitor how these goals are achieved. A portfolio encourages constant self reflection. PMID- 11895237 TI - Does observation add to the validity of the long case? PMID- 11895239 TI - Building collegiality: the real value of problem-based learning. PMID- 11895238 TI - A schematic approach to diagnosing and resolving lecturalgia. AB - BACKGROUND: The lecture is a much used and much criticized teaching method. Lecturalgia (painful lecture) is a frequent cause of morbidity for both teachers and learners. The etiology of lecturalgia is multifactorial and multiple lecturing pathologies frequently coexist. The 'Clinical Presentation' curriculum at the University of Calgary encourages the use of 'schemes' that provide a scaffolding for learning and a starting point for approaching (clinical) problems. Thus far this approach has not been used to tackle teaching or learning problems. AIM: Our aim in this paper was to devise a schematic approach to diagnosing lecturing problems and to make evidence-based recommendations on how to resolve lecturing problems. We have suggested that causes of lecturalgia can be divided into three categories: poor judgement; poor organization; and poor delivery. Our proposed scheme is based upon these three categories that are then subcategorized. RESULTS: We have reviewed the medical education literature in an attempt to provide evidence-based recommendations for the remediation of lecturing problems within each subcategory. CONCLUSION: Where trial evidence is lacking we have made recommendations that are consistent with cognitive theory or expert opinion. Finally, where expert opinion does not exist, we have taken the liberty (literary license) of providing nonexpert opinion! PMID- 11895240 TI - Tales from tooting: reflections on the first year of the MBBS graduate entry programme at St George's Hospital Medical School. PMID- 11895241 TI - Students of the past...doctors of the future. PMID- 11895242 TI - Imaginative writing. PMID- 11895243 TI - Why does so much depend upon a red wheelbarrow? What is the point of a medical writing option? PMID- 11895244 TI - Learning to look: developing clinical observational skills at an art museum. AB - CONTEXT: Clinical diagnosis involves the observation, description, and interpretation of visual information. These skills are also the special province of the visual arts. We describe an educational collaboration between a medical school and an art museum, designed for the purpose of developing student skills in observation, description, and interpretation. OBJECTIVES: In the programme, medical students first examine painted portraits, under the tutelage of art educators and medical school faculty. Then, the students examine photographs of patients' faces and apply the same skills. CONCLUSION: This programme, well received by students and faculty, appeared to help the students not only in improving their empirical skills in observation, but also in developing increased awareness of emotional and character expression in the human face. PMID- 11895245 TI - Internationalization of undergraduate medical studies: promoting clinical tourism or academic development? PMID- 11895246 TI - Summer in Durham: abroad thoughts from home. PMID- 11895247 TI - The value of silence. PMID- 11895248 TI - Managed care. PMID- 11895249 TI - Competence and the laying of blame. PMID- 11895250 TI - The assessment of poorly performing doctors: the development of the assessment programmes for the General Medical Council's Performance Procedures. AB - BACKGROUND: Modernization of medical regulation has included the introduction of the Professional Performance Procedures by the UK General Medical Council in 1995. The Council now has the power to assess any registered practitioner whose performance may be seriously deficient, thus calling registration (licensure) into question. Problems arising from ill health or conduct are dealt with under separate programmes. METHODS: This paper describes the development of the assessment programmes within the overall policy framework determined by the Council. Peer review of performance in the workplace (Phase 1) is followed by tests of competence (Phase 2) to reflect the relationship between clinical competence and performance. The theoretical and research basis for the approach are presented, and the relationship between the qualitative methods in Phase 1 and the quantitative methods in Phase 2 explored. CONCLUSIONS: The approach is feasible, has been implemented and has stood legal challenge. The assessors judge and report all the evidence they collect and may not select from it. All their judgements are included and the voice of the lay assessor is preserved. Taken together, the output from both phases forms an important basis for remediation and training should it be required. PMID- 11895251 TI - The General Medical Council's Performance Procedures: the development and implementation of tests of competence with examples from general practice. AB - OBJECTIVE: This paper describes the development of the tests of competence used as part of the General Medical Council's assessment of potentially seriously deficient doctors. It is illustrated by reference to tests of knowledge and clinical and practical skills created for general practice. SUBJECTS AND TESTS: A notional sample of 30 volunteers in 'good standing' in the specialty (reference group), 27 practitioners referred to the procedures and four practitioners not referred but who were the focus of concern over their performance. Tests were constructed using available guidelines and a specially convened working group in the specialty. METHODS: Standards were set using Angoff, modified contrasting group and global judgement methods, as appropriate. RESULTS: Tests performed highly reliably, showed evidence of construct validity, intercorrelated at appropriate levels and, at the standards employed, demonstrated good separation of reference and referred groups. Likelihood ratios for above and below standard performance based on competence were large for each test. Seven of 27 doctors referred were shown not to be deficient in both phases of the performance assessment. PMID- 11895252 TI - Training the assessors for the General Medical Council's Performance Procedures. AB - From July 1997, the General Medical Council (GMC) has had the power to investigate doctors whose performance is considered to be seriously deficient. Assessment procedures have been developed for all medical specialties to include peer review of performance in practice and tests of competence. Peer review is conducted by teams of at least two medical assessors and one lay assessor. A comprehensive training programme for assessors has been developed that simulates the context of a typical practice-based assessment and has been tailored for 12 medical specialties. The training includes the principles of assessment, familiarization with the assessment instruments and supervised practice in assessment methods used during the peer review visit. High fidelity is achieved through the use of actors who simulate third party interviewees and trained doctors who role play the assessee. A subgroup of assessors, selected to lead the assessment teams, undergo training in handling group dynamics, report writing and in defending the assessment report against legal challenge. Debriefing of assessors following real assessments has been strongly positive with regard to their preparedness and confidence in undertaking the assessment. PMID- 11895253 TI - The reproducibility of assessing radiological reporting: studies from the development of the General Medical Council's Performance Procedures. AB - OBJECTIVES: To investigate the reproducibility of peer ratings of consultant radiologists' reports, as part of the new General Medical Council (GMC) Performance Procedures. DESIGN: An evaluation protocol was piloted, used in a blocked, balanced, randomized generalizability analysis with three blocks of three judges (raters), each rating 30 reports from 10 radiologists, and re-rated to estimate intrarater reliability with conventional statistics (kappa). SETTING: Rating was performed at the Royal College of Radiologists. Volunteers were sampled from 23 departments of radiology in university teaching and district general hospitals. PARTICIPANTS: A nationally drawn non-random sample of 30 consultant radiologists contributing a total of 900 reports. Three trained and six non-trained judges were used in the rating analysis. RESULTS: A protocol was generated that was usable by judges. Generalizable results would be obtained with not less than three judges all rating the same 60 reports from a radiologist. CONCLUSIONS: Any assessment of performance of technical abilities in this field will need to use multiple assessors, basing judgements on an adequate sample of reports. PMID- 11895254 TI - Senior health professional's perceptions of variations in medical practice: a qualitative and quantitative study. AB - Following the introduction of the UK General Medical Council's regulations on the handling of poor medical performance, an interview and survey study was carried out among senior health professionals in the National Health Service (NHS). The aims of the study were to explore the respondents' perceptions of poor medical performance and to seek their experience of handling poorly performing doctors. Sixteen interviews were held face to face, followed by 28 telephone interviews. Subsequently, using similar questions to those in the interview schedule, a survey was carried out among senior health professionals across the NHS. Interview results identified a number of barriers to resolving poor performance, such as the unwillingness of some doctors to seek advice and the protective culture which prevented complaints being made against doctors. Survey respondents had high standards by which they judged poor performance, but they were more hesitant about considering poor consultation skills as being of the same significance as poor technical skills. However, problems with communication skills were the most frequently reported type of poor performance. The new arrangements for handling NHS doctors whose performance is perceived to be poor have much to do to overcome the barriers to effective action expressed by the respondents in this study. PMID- 11895255 TI - Good Medical Practice: comparing the views of doctors and the general population. AB - OBJECTIVES: To assess the views of the general public and general practitioners (GPs) on the General Medical Council's Good Medical Practice. METHODS: A large national consumer survey organization (MORI) was commissioned to conduct a survey of a representative sample of the general public using quota sampling methods with randomly selected sampling points across Great Britain. A similar survey was carried out of GPs across Great Britain. Questions were asked of the general public about their perceptions on medical practitioners keeping their knowledge sufficiently up to date to do their job, on complaints being properly investigated, and about doctors being removed from the General Medical Council register if they failed to perform a specifically defined duty. The GPs were asked the same questions about duties. The general public was also asked whether they worked in the National Health Service (NHS), about any recent experiences as a patient in the NHS and whether they had ever complained about a doctor. RESULTS: A total of 1949 people were interviewed in their homes along with 199 GPs. Over one-third either worked in the NHS or had friends or relatives who worked in the NHS. Eleven per cent felt they had reason to complain about a doctor although only 6% had made a formal or informal written or verbal complaint. The general public and GPs held similar views on reasons for doctors being removed from the register, although the public's threshold was somewhat lower than the doctors. CONCLUSIONS: GPs and the general public have similar views on good medical practice, although the degree of concern shown by the public is greater than that registered by doctors. Both groups show similar patterns of variation in their perception of the importance of some duties compared with others. PMID- 11895256 TI - UK doctors' attitudes to the General Medical Council's Performance Procedures, 1997-99. AB - OBJECTIVES: The UK General Medical Council's Performance Procedures were introduced in 1997. This study aimed to assess the changing knowledge and attitudes about the procedures in British doctors at the time of their introduction and in the following 2 years. METHODS: Three questionnaire surveys, of separate representative samples of 800 UK doctors, were carried out in November of 1997, 1998 and 1999. The surveys assessed awareness of Good Medical Practice, attitudes to the Performance Procedures, agreement with Duties of a Doctor as a basis for disciplinary procedures, and attitudes to the Performance Procedures. RESULTS: Although awareness of the procedures increased over the period 1997-99, there was no concurrent increase in agreement with the core principles of the procedures, the Duties of a Doctor, which are spelled out in Good Medical Practice. Of 12 separate attitudes to the procedures, changes were found in eight over the time period, all but two of which were negative, and not in support of the procedures. Nevertheless many doctors were changing their practice as a result of the procedures, and that proportion increased during the period 1997-99. CONCLUSIONS: Although doctors became more aware of the procedures, that increasing awareness was not accompanied by an increasing agreement with the procedures' underlying principles or their wider implications. PMID- 11895257 TI - Good medical practice: the duties of a doctor registered with the General Medical Council. PMID- 11895258 TI - The General Medical Council's Performance Procedures: peer review of performance in the workplace. AB - The General Medical Council procedures to assess the performance of doctors who may be seriously deficient include peer review of the doctor's practice at the workplace and tests of competence and skills. Peer reviews are conducted by three trained assessors, two from the same speciality as the doctor being assessed, with one lay assessor. The doctor completes a portfolio to describe his/her training, experience, the circumstances of practice and self rate his/her competence and familiarity in dealing with the common problems of his/her own discipline. The assessment includes a review of the doctor's medical records; discussion of cases selected from these records; observation of consultations for clinicians, or of relevant activities in non-clinicians; a tour of the doctor's workplace; interviews with at least 12 third parties (five nominated by the doctor); and structured interviews with the doctor. The content and structure of the peer review are designed to assess the doctor against the standards defined in Good Medical Practice, as applied to the doctor's speciality. The assessment methods are based on validated instruments and gather 700-1000 judgements on each doctor. Early experience of the peer review visits has confirmed their feasibility and effectiveness. PMID- 11895259 TI - Another mirror shattered? Tobacco industry involvement suspected in a book which claims that nicotine is not addictive. PMID- 11895260 TI - Pattern of alcohol drinking and stroke: a comment on Mazzaglia et al. PMID- 11895261 TI - Alcohol consumption and stroke--the difficulties in giving responsible advice. PMID- 11895262 TI - Alcohol and stroke: how generalizable are the current research findings? PMID- 11895263 TI - No positive tests for syphilis in 6 years of observation among heroin drug users in north-eastern Italy. PMID- 11895264 TI - Average volume of alcohol consumption, patterns of drinking and mortality among young Europeans in 1999. PMID- 11895265 TI - Dutch cannabis policy evaluated. PMID- 11895266 TI - Alcohol problems in Ecuador: prevention activities. PMID- 11895267 TI - Smoking as a risk factor for Alzheimer's disease: contrasting evidence from a systematic review of case-control and cohort studies. AB - AIMS: To investigate the risk of Alzheimer's disease (AD) associated with smoking. DESIGN: Meta-analyses of case-control and cohort studies. DATA SOURCE: Index Medicus-Medline (1966-April 2000) and PsycINFO (1984-April 2000) databases were systematically consulted for the retrieval of references. This search was supplemented by manual search of relevant references quoted by other studies and reviews. STUDY SELECTION: Irrelevant abstracts and articles were identified by one of the authors. These papers were retrieved and examined by at least two of the authors, who initially assessed them for the relevance of the exposure (smoking), outcome (AD) and study-design (case-control or cohort study). DATA EXTRACTION: Two reviewers rated independently the quality of selected papers. Whenever possible, raw data were extracted and the crude odds ratio (OR) calculated using the Cornfield method. The pooled risk ratios were estimated using a fixed-effects model. FINDINGS: Twenty-one case-control studies reported data on 5323 subjects. The estimated pooled odds ratio (OR) was 0.74 [95% confidence interval (CI) = 0.66-0.84]. In another analysis incorporating ORs adjusted for confounding variables (such as age, sex, schooling and alcohol use), the pooled odds ratio was 0.82 (95% CI = 0.70-0.97). Finally, in a analysis that included only the four case-control studies that used matched design the pooled odds ratio was 0.82 (95% CI = 0.53-1.27). Eight cohort studies reported data on 43 885 people at risk-the overall relative risk (RR) of AD among ever smokers was 1.10 (95% CI = 0.94-1.29). Restricting the analysis to the two cohort studies that described the number of subjects who were smokers at baseline and later developed AD produced a RR of 1.99 (95% CI = 1.33-2.98). CONCLUSIONS: Case control and cohort studies produce conflicting results as to the direction of the association between smoking and AD. Survival bias and other methodological problems associated with case-control studies may partly explain this difference. Access to information collected by ongoing follow-up studies may contribute to clarify the role of smoking in AD. If new results confirm that smoking is associated with increased risk of AD, then smoking prevention and cessation should become public health priorities in the fight against dementia. PMID- 11895268 TI - Effects of depression and social integration on the relationship between alcohol consumption and all-cause mortality. AB - AIMS: This study was designed to assess the potentially confounding influences of social integration and depression on the form of the relationship between alcohol consumption and all-cause mortality. DESIGN, PARTICIPANTS AND MEASUREMENT: Respondents from the 1984 US National Alcohol Survey (N = 5177) were followed by searching the National Death Index (NDI) through 1995; 540 were identified as deceased. Predictor variables in a Cox proportional hazards model included gender, ethnicity, marital status, income, smoking, age and alcohol consumption (volume and patterns). Two social variables and their interactions with alcohol consumption were added, the Center for Epidemiological Studies Depression (CES-D) scale and an eight-item social isolation scale. FINDINGS: The J-shaped risk curve for all-cause mortality by volume was approximated for men but not significantly for women. In addition heavy drinking occasions independently contributed to mortality in men. Low social integration (bottom 12%) had no significant effects on mortality or on the relationship between alcohol consumption and mortality curve. Inclusion of the interaction between alcohol consumption and depression proved significant for heavy male drinkers (> six drinks on average per day) and for female former drinkers with heavy drinking occasions. In both cases, the respective subgroup, which additionally was depressed, had about four times the risk of a life-time abstainer. CONCLUSIONS: The relationship of alcohol consumption to 11-year all-cause mortality in a general population indicated little confounding effect of social isolation, but revealed important interactions with depression for heavy male drinkers and heavy female ex drinkers. PMID- 11895269 TI - A prospective study of mortality among drug misusers during a 4-year period after seeking treatment. AB - AIMS: The opportunity to study deaths as they occur within the framework of a prospective cohort study is relatively uncommon. This study investigates deaths among drug misusers over a 4-year period, with specific attention to the circumstances and causes of death, and risk factors for mortality. The study also critically examines the recording of drug-related deaths. DESIGN, SETTING, PARTICIPANTS: Prospective cohort study of 1075 drug misusers recruited to 54 treatment programmes during 1995. MEASUREMENTS: Data derived from interviews conducted with clients at intake, death certificates and post-mortem examinations. FINDINGS: The annual mortality rate was 1.2%, about six times higher than that for a general, age-matched population. Fourteen per cent of the deaths were due to self-inflicted injuries, accidents or violence and 18%, were due to medical causes. The majority of deaths (68%) were associated with drug overdoses. Opiates were the drugs most commonly detected during post-mortem examinations. In the majority of cases, more than one drug was detected. Polydrug use and, specifically, heavy drinking, and use of benzodiazepines and amphetamines, were identified as risk factors for mortality. Anxiety and homelessness were also predictive of increased mortality. CONCLUSIONS: We suggest that drug misusers and those working with drug misusers need to be more alert to the risks of polydrug use, including the combined use of alcohol with illicit drugs. The study revealed inconsistencies in the recording of drug-related deaths on death certificates. The routine recording of all substances detected during toxicological examination would improve the accuracy of death certification. PMID- 11895270 TI - Alpha2-adrenergic agonists in opioid withdrawal. AB - OBJECTIVES: This paper presents the main findings of a systematic (Cochrane) review of the effectiveness of alpha2-adrenergic agonists in managing opioid withdrawal. DESIGN: The original systematic review included controlled trials that compared alpha2-adrenergic agonists with another form of treatment (or placebo) in participants who were primarily opioid-dependent. MAIN FINDINGS: Ten studies compared a treatment regime based on an alpha2-adrenergic agonist with one based on reducing doses of methadone. Withdrawal intensity is similar to, or marginally greater with alpha2-adrenergic agonists, but signs and symptoms of withdrawal occur and resolve earlier in treatment. Participants stay in treatment longer with methadone. The likelihood of completing withdrawal is similar, or slightly less, with clonidine or lofexidine. Clonidine is associated with more adverse effects than reducing doses of methadone. Three studies compared the alpha2-adrenergic agonists, clonidine and lofexidine. Lofexidine does not reduce blood pressure to the same extent as clonidine, but is otherwise similar to clonidine. CONCLUSIONS: Participants stay in treatment longer with methadone regimes, which may provide greater opportunity for psychosocial intervention. Methadone regimes may be preferable for withdrawal in outpatient settings where the risk of relapse to heroin use is high. The use of methadone may also facilitate transfer to maintenance treatment should completion of withdrawal become unlikely. For those who are well prepared for withdrawal and seeking earlier resolution of withdrawal symptoms, alpha2-adrenergic agonist treatment may be preferred. Clonidine and lofexidine appear equally effective for inpatient settings, but the lower incidence of hypotension makes lofexidine more suited to use in outpatient settings. PMID- 11895271 TI - Acceptability and availability of pharmacological interventions for substance misuse by British NHS treatment services. AB - AIMS: Despite their potential advantages, many of the pharmacological interventions available to treat substance misuse are controversial and their acceptability within the United Kingdom (and other countries) has only recently begun to be investigated. DESIGN: A questionnaire mailed to British National Health Service (NHS) alcohol and drug treatment services asked respondents to rate the acceptability and availability of 11 pharmacological interventions for substance misuse employed to relieve withdrawal, reduce the likelihood of relapse and opiate overdose and substitute pharmaceuticals for illicit drugs. PARTICIPANTS: A sample of NHS substance misuse services (n = 265) listed in one or more directories of services in England, Wales and Scotland. FINDINGS: Substitute methadone for opiate addiction, substitute benzodiazepines for benzodiazepine-dependent patients, lofexidine for opiate detoxification, naltrexone for opiate relapse prevention and acamprosate for alcohol relapse prevention were widely acceptable and available interventions. Another subset of medications-buprenorphine for opiate detoxification, take-home naloxone for overdose prevention and substitute prescribing of levo-alpha-acetyl-methadol (LAAM), heroin and dexamphetamine-garnered less support, but the majority of participants rated even these therapies as acceptable. Ultra-rapid detoxification under sedation was the intervention rated as least acceptable to, and was one of the two least frequently available from, responding NHS services. CONCLUSIONS: Differences among specific medications notwithstanding, a wide range of harm reduction and abstinence-orientated interventions were acceptable to and available from NHS services. Acceptance and availability are probably limited by a combination of practical, economic, safety, efficacy and theoretical considerations. PMID- 11895272 TI - The effects of drug abuse prevention at school: the 'Healthy School and Drugs' project. AB - AIMS: To examine the effects of the 'Healthy School and Drugs' project, a Dutch school-based drug prevention project that was developed in the late 1980s and disseminated during the 1990s. This programme is currently being used by 64-73% of Dutch secondary schools and it is estimated that at least 350000 high school students receive this intervention each year. DESIGN, SETTING AND PARTICIPANTS: A quasi-experimental study in which students of nine experimental (N = 1156) schools were compared with students of three control schools (N = 774). The groups were compared before the intervention, 1 year later, 2 years later and 3 years later. MEASUREMENTS: Self-report measures of tobacco, alcohol and marijuana use, attitudes towards substance use, knowledge about substances and self efficacy. FINDINGS: Some effects on the use of tobacco, alcohol and cannabis were found. Two years after the intervention, significant effects could still be shown on alcohol use. Effects of the intervention were also found on knowledge, but there was no clear evidence for any effects on attitude towards substance use and on self-efficacy. CONCLUSIONS: This study shows the Healthy School and Drugs project as implemented in Holland may have some effect on drug use in the children exposed to it. PMID- 11895273 TI - Risk factors for symptom exacerbation among treated patients with substance use disorders. AB - AIMS: The purpose of this study was to develop an index of risk factors to identify patients prospectively with substance use disorders whose substance use symptoms exacerbate during or shortly after treatment, and to identify characteristics of care that may reduce the likelihood of exacerbation. DESIGN, SETTING, PARTICIPANTS: On the basis of data obtained from a nation-wide outcomes monitoring system, a group of 2809 treated patients experienced an exacerbation of their substance use symptoms. These patients were matched on baseline substance abuse problems with 5618 patients who remained stable or improved. MEASUREMENTS AND FINDINGS: Risk factors for substance use symptom exacerbation included younger age, non-married status and residential instability; long-term use of drugs, prior arrests, prior alcohol treatment, alcohol and drug abuse or dependence diagnoses, cocaine abuse or dependence and more severe self-rated drug problems; and psychiatric problems. High-risk patients who obtained a longer episode of mental health care were less likely to experience an exacerbation of symptoms. CONCLUSIONS: Clinicians can identify at treatment entry patients whose substance use symptoms are likely to exacerbate and, by providing these patients a longer duration of care, may reduce the likelihood of symptom exacerbation. PMID- 11895274 TI - The urge to smoke depends on the expectation of smoking. AB - AIMS: An earlier study (Dols et al. 2000) suggested that cue-induced urge to smoke depends on the expectation of smoking. The present study tried to replicate the findings under stringently controlled conditions. DESIGN: A 2 (context) x 2 (cues) x 6 (trial) within-subject design. Each smoker entered two different contexts; one context predicted the future occurrence of smoking (i.e. one puff of a cigarette) and one context predicted the non-occurrence of smoking. In each context smokers were exposed to smoking cues (i.e. cigarettes and lighter) or not. SETTING: Laboratory at Maastricht University. PARTICIPANTS: Thirty-two daily smokers, smoking at least five cigarettes a day for at least 2 years. MEASUREMENTS: Participants reported their urge to smoke in each context in the presence and absence of smoking cues using a computerized visual analogue scale (VAS). FINDINGS: The results revealed that the urge to smoke was higher in a context in which smoking was expected relative to a context in which it was not expected. As in the previous study the urge-inducing effect of smoking cues was larger in the smoking context than in the non-smoking context. Moreover, smoking cues did not have a significant effect in the non-smoking context. CONCLUSIONS: It was shown that smoking cues elicit craving due mainly to a generated expectation of the occurrence of smoking and less due to salience or long history of associative learning. Theoretical and practical implications of the results are discussed. PMID- 11895275 TI - The Brazilian Association for the Study of Alcohol and Other Drugs. AB - The Brazilian Association for the Study of Alcohol and other Drugs [Associacao Brazileira de Estudos do Alcool e outras Drogas (ABEAD)] is a non-profit entity with the purpose of encouraging discussions and exchanges on advances in the prevention, treatment and research of addition. It currently brings together 840 professionals from different areas, such as Psychiatry, Social Assistance, Nursing, Psychology, Sociology, Law, in addition to community leaders, thus having an innovative and dynamic character. It organises annual national congresses and regional meetings hosting foreign guests. The society has a style that is innovative and dynamic. After more than 20 years of work, ABEAD exerts significant national influence and its members have been invited to participate in the main decisions regarding issues linked to drug abuse in Brazil. Among other achievements, it has contributed to the proposals on prevention and treatment policies and the standardization of the different levels of assistance provided by public health-care. In addition, its members have worked closely with the initiatives made by the government and medical entities in the search for a consensus on the treatment of the different types of dependence, including alcohol dependence and smoking. ABEAD's goal for the future is to broaden its participation in the national drug policy arena and to intensify the work with other medical organizations and with the community. PMID- 11895276 TI - Another 'gold standard' shattered? Re-opening the 'done deal' of conflict of interest disclosure. PMID- 11895277 TI - The effect of removable partial dentures on periodontal health of abutment and non-abutment teeth. AB - BACKGROUND: A removable partial denture (RPD) is a common treatment available for restoration of partially edentulous ridges. Longitudinal studies indicate that RPDs have been associated with increased gingivitis, periodontitis, and abutment mobility. METHODS: A total of 205 patients with RPDs participated in this study. There were 80 males and 125 females aged 38 to 89, with 123 maxillary and 138 mandibular RPDs. Patients were wearing existing RPDs for different periods ranging from 1 to 10 years. A two-part questionnaire was devised for this study. In the first part, patients answered questions on gender; age; smoking habits; denture age; denture wearing habits; mouth odor; and problems with food accumulating under the denture base, on the outside surface of the denture, and on the outside surface of remaining teeth after eating. The Kennedy classification, material, denture support, denture base shape, and number of teeth in contact, number of existing clasps, and occlusal rests were categorized. The quality of denture construction was also evaluated. In the second part of the questionnaire, baseline recordings of plaque (PI), gingival (GI), and calculus (CI) indexes were made, and Tarbet index (TI), as well as probing depth (PD), gingival recession (GR), and tooth mobility (TM) were measured, both on abutment and non-abutment teeth. RESULTS: Significant differences (P <0.01) were noted for PI, Cl, GI, PD, TM, and GR between abutment and non-abutment teeth, with abutment teeth showing more disease. CONCLUSIONS: RPD design plays an important role in the state of the periodontium. Appropriate design and good oral hygiene may decrease the appearance of periodontal disease. PMID- 11895278 TI - Cytokine regulation of gingival fibroblast lysyl oxidase, collagen, and elastin. AB - BACKGROUND: Systemic therapy with cyclosporin A, phenytoin, and nifedipine modulates cytokine levels in human gingival tissues. Functional relationships between altered cytokine levels and gingival extracellular matrix production are partially characterized. The present study investigates in cultured human gingival fibroblasts the regulation of lysyl oxidase, alpha-1 type I collagen, and elastin by selected cytokines that are elevated in drug-induced gingival overgrowth tissues. METHODS: Normal human gingival fibroblasts were cultured and then treated with selected cytokines: interleukin (IL)-1beta, IL-6, platelet derived growth factor (PDGF)-BB, and basic fibroblast growth factor (bFGF or FGF 2). Cells were harvested at intervals, and changes in lysyl oxidase enzyme activity, and in mRNA levels of lysyl oxidase, alpha-1 type I collagen, and elastin were determined. RESULTS: bFGF reproducibly and significantly decreased human gingival fibroblast lysyl oxidase and alpha-1 type I collagen mRNA levels in a dose- and time-dependent manner; 1 nM bFGF reduced lysyl oxidase and collagen mRNA levels to 53% and to less than 10% of control after 48 hours of treatment. Interestingly, bFGF downregulated lysyl oxidase enzyme activity by 10% to 20%. IL-1, IL-6, and PDGF-BB did not significantly regulate lysyl oxidase enzyme activity, or alpha-1 type I collagen, elastin, and lysyl oxidase mRNA levels under the conditions tested. CONCLUSIONS: Previous studies have shown that modulated levels of bFGF occur in gingiva as a result of certain pharmacologic therapies. The present study suggests that modulated levels of bFGF likely influence gingival connective tissue metabolism. PMID- 11895279 TI - Smoking and complications of endosseous dental implants. AB - BACKGROUND: The purpose of this study was to compare the incidence of the complications and survival rate related to dental implants among smokers and non smokers, and to evaluate the influence of smoking by analyzing data of 959 implants placed in 261 patients during the years 1995 to 1998. METHODS: Patients were divided into 3 groups: non-smokers, mild smokers (up to 10 cigarettes per day) and heavy smokers (more than 10 cigarettes per day); smokers were divided into 2 subgroups according to duration of smoking (less or more than 10 years). Complications included minor (spontaneous implant exposure), major (spontaneous implant exposure requiring surgical intervention), and implant failure. The influence of smoking was analyzed for the type of implant cover screw and immediate versus late implantation. RESULTS: The overall failure rate was 2% for non-smokers and 4% for all smokers. Minor and major complications were found in higher percentages (46%) in the smoking groups than in the non-smoking group (31%). A significantly higher incidence of complications was found among smokers who received dental implants with high cover screws (63%) compared to those who received dental implants with flat cover screws (27%). CONCLUSIONS: This study establishes a relationship between implant complications and smoking, implant type (external or internal hex), and time of implantation as significant factors. A higher incidence of complications was found in the smoking group, especially in implants that had a high cover screw. Most complications will not lead to failures. Immediate implants failed less frequently than non-immediate implants. Limiting or reducing smoking habits will decrease complications of endosseous dental implants. PMID- 11895280 TI - Combined periodontal regenerative technique in human intrabony defects by collagen membranes and anorganic bovine bone. A controlled clinical study. AB - BACKGROUND: Combined periodontal regenerative technique (CPRT) is a surgical procedure that combines the use of barrier membranes with a filling material in the treatment of periodontal defects. The effectiveness of CPRT has been evaluated in many studies in comparison to GTR with membranes alone, but conflicting results have been obtained by different clinicians, particularly in the treatment of intrabony defects. The aim of the present study was to compare CPRT to GTR with collagen membranes in the treatment of human intrabony defects characterized by a relevant 1-wall component. METHODS: Thirty-four (34) healthy, non-smoking patients affected by moderate to severe chronic periodontitis participated in this study. Each patient had good oral hygiene and at least 1 radiographically detectable intrabony defect > or = 4 mm, with a 1-wall component of at least 50% of the defect, involving 2 tooth surfaces or more with a probing depth (PD) > or = 6 mm. Seventeen (17) subjects were randomly assigned to the test group and underwent CPRT by anorganic bovine bone and a collagen membrane, and 17 randomly assigned to the control group who received GTR with a collagen membrane alone. Pre- and post-therapy clinical parameters (probing depth [PD]; clinical attachment level [CAL]; gingival recession [GR]) and intrasurgical parameters (depth of intraosseous component [IOC]; level of the alveolar crest [ACL]) were compared between test and control groups 1 year after treatment. Vertical bone gain (VBG) from the base of the defect to the cemento-enamel junction was also evaluated in both groups. RESULTS: At the 1-year examination, clinical and intrasurgical parameters showed statistically significant changes within each experimental group from baseline. A statistically greater CAL gain was reported in the test group (P<0.05), whereas the control group exhibited more GR and alveolar crest resorption at a statistically significant level (P<0.01). VBG was significantly greater (P<0.01) at test sites (5.23 +/- 1.30 mm) compared to controls (3.82 +/- 1.28 mm). CONCLUSIONS: The results suggest that the use of CPRT may be preferred when bioabsorbable membranes are used to treat intrabony defects characterized by unfavorable architecture. PMID- 11895281 TI - Comparison of conventional periodontal maintenance versus scaling and root planing with subgingival minocycline. AB - BACKGROUND: Alternative regimens using subgingival antimicrobials compared to conventional periodontal maintenance (PM) may lead to more efficient protocols. The purpose of this study was to evaluate treatment time and clinical and radiographic outcomes in 2 periodontitis cohorts, one receiving conventional PM and the other receiving scaling and root planing (SRP) and multiple doses of subgingival minocycline. METHODS: Moderate to advanced chronic periodontitis patients were concurrently treated with either: 1) scaling and root planing and 4 subgingival doses of minocycline microspheres in all > or = 5 mm pockets over a 6 month period (RP/M; n = 24 patients); or 2) conventional 3-month periodontal maintenance (PM; n = 24 patients). Clinical and radiographic measurements, including probing depth (PD), clinical attachment level (CAL), and interproximal bone height (BH), were analyzed in 2 premolar/molar interproximal > or = 5 mm pockets at baseline and 1 year using paired t tests, analysis of variance, chi square analysis, and correlation coefficients. RESULTS: Baseline clinical and radiographic data were similar between RP/M and PM patients. Probing depths showed greater mean improvement in RP/M (0.9 +/- 0.1 versus 0.4 +/- 0.1 mm, P = 0.02), with 25% of subjects in RP/M gaining > or = 2 mm compared to 4.2% in PM (differences were statistically significant). The mean loss in bone height and percent subjects losing bone height were less in RP/M (0.05 +/- 0.05 mm; 12.5%) than PM (0.09 +/- 0.08 mm; 16.7%), but bone height differences were not statistically significant. A subset of RP/M molar furcation sites responded with similar PD reduction and no BH loss over 1 year. While cross-sectional RP/M data between CAL and BH, or PD and CAL were highly correlated, changes over 1 year were not correlated among any of these parameters. CONCLUSIONS: Scaling and root planing and subgingival minocycline in experimental sites took little time (<5 minutes/appointment), but resulted in more probing depth reduction and less frequent bone height loss than conventional periodontal maintenance. PMID- 11895282 TI - The effects of periodontal therapy on intracrevicular prostaglandin E2 concentrations and clinical parameters in pregnancy. AB - BACKGROUND: The increase in circulating levels of progesterone during pregnancy stimulates production of prostaglandins, especially prostaglandin E2, possibly resulting in pregnancy gingivitis. The purpose of this study is to evaluate the influence of prostaglandin E2 concentrations on gingival tissues in pregnancy and to assess its relationship to clinical parameters. METHODS: This study evaluates the effects of periodontal treatment on clinical indices including plaque index, gingival index, probing depth, and gingival crevicular fluid prostaglandin E2 levels of 22 pregnant women in their first, second, and third trimesters. Initial periodontal therapy consisting of scaling, root planing, and oral hygiene instruction was performed at the beginning of the first trimester and repeated each trimester. Prostaglandin E2 concentrations in gingival crevicular fluid were determined using a commercially available enzyme immunoassay kit. The statistical tests used were paired sample test and correlation analysis. RESULTS: The results of the study show that periodontal therapy has resulted in an improvement in clinical parameters (P<0.05). There is also a statistically significant decrease in levels of prostaglandin E2 at the second and third trimesters following periodontal therapy (P <0.001). The correlation between prostaglandin E2 concentrations and clinical parameters is found to be non-significant (P >0.05). CONCLUSIONS: Our data indicate that levels of prostaglandin E2 in gingival crevicular fluid may be used as a marker of gingival inflammation in order to determine the effects of periodontal therapy in pregnancy. Periodontal therapy that is performed throughout the entire pregnancy period may help prevent the threat of pregnancy gingivitis. PMID- 11895283 TI - The effect of sociocultural status on periodontal conditions in pregnancy. AB - BACKGROUND: The effects of sociocultural status on periodontal conditions in pregnant women have been reported by a number of researchers and there have been speculations about the effects of hormonal changes, patients' systemic health, and socio-cultural characteristics on periodontal health during pregnancy. METHODS: This study evaluates the periodontal condition of 61 pregnant women at their first, second, and third trimesters, and the relation between the demographic (age, professional level, education) and clinical variables (previous periodontal care, frequency of tooth brushing). The clinical indices, including plaque index, gingival index, and probing depth measurements were repeated at the first, second, and third trimesters. The statistical tests used were stepwise analysis and paired sample test. RESULTS: The results of the study showed that the plaque index, gingival index, and probing depth scores increased gradually in the first, second, and third trimesters, although oral hygiene instructions were given to the entire study population. The level of statistical significance was established at P <0.05. When the clinical parameters and demographic variables were compared, only educational level and periodontal care seemed to be statistically significant (P<0.05). CONCLUSIONS: Our clinical index scores were related to the educational level of the study population. When the educational level of the study group decreased, the plaque, gingival index, and probing depth scores contrarily increased. Also non-attendance for previous periodontal care increased the scores of plaque index and probing depth. In view of the results of our study, it might be suggested that simple preventive oral hygiene programs may help maintain healthy gingiva during pregnancy. PMID- 11895284 TI - Dentin morphology and permeability after brushing with different toothpastes in the presence and absence of smear layer. AB - BACKGROUND: The purpose of this study was to evaluate the morphology and permeability of dentin after brushing with different toothpastes in the presence and absence of smear layers. METHODS: Dentin discs were prepared from extracted third molars. Dentin permeability was measured using a hydraulic pressure apparatus working at 70 cm H2O pressure. Dentin was treated with 0.5 M EDTA for 5 minutes and washed to remove a smear layer and to establish the maximum permeability of each dentin disc, which was expressed as 100%. A new smear layer was then created on the upper surface using a #400 carbide paper under water for 30 seconds. Dentin permeability of the smear layer-covered dentin was measured and expressed as a percentage of the maximum permeability of that specimen, permitting each specimen to serve as its own control. Each sample was then brushed by a mechanical device under water for 3 minutes with constant pressure of 250 g using a medium toothbrush and permeability remeasured. Finally, 1 of 5 different toothpastes was applied on the dentin and brushed for 3 minutes. In another group, the same procedures were performed with the exception of the smear layer production, so it was possible to calculate changes in the permeability of dentin with open tubules following brushing. Scanning electron microscopic (SEM) examination of dentin was obtained before and after treatments with brushing and toothpastes. RESULTS: Dentin permeability was reduced by brushing procedures when the smear layer was absent, but it was increased when the smear layer was present. Toothpaste application reduced dentin permeability when no smear layer was present on the top of the surface, but modified and increased permeability of samples with smear layers. SEM observations demonstrated the presence of dentifrice particles on dentin surfaces and inside dentinal tubules, and this may be responsible for the observed reductions in permeability. Smear plugs produced during dentin brushing were not removed by the dentifrices. CONCLUSIONS: Dentin permeability and morphology are significantly affected by toothbrushing and by the type of dentifrice used. The presence of smear plugs in the dentin may decrease severity. PMID- 11895285 TI - Evaluation of pluronic polyols as carriers for grafting materials: study in rat calvaria defects. AB - BACKGROUND: Pluronic polyols are a family of non-ionic surfactants currently used as drug carriers for antibiotic, anti-inflammatory, and anti-neoplastic agents. Therapeutic administration of non-ionic surface-active agents is known to facilitate early collagen synthesis and microcirculation, thus promoting wound healing. The purpose of this study was to determine the in vivo effects of pluronic polyols combined with either an allograft or an alloplast on the healing of critical-sized calvarial defects. METHODS: One hundred fifty (150) adult (95 to 105 days old) male Sprague-Dawley rats weighing between 375 and 425 g were randomly and evenly assigned to each of 15 separate treatment groups and anesthetized, and 8 mm calvarial critical-sized defects were created. Pluronic F 68 (F-68) or pluronic F-127 (F-127) was administered either topically or systemically and in conjuction with demineralized bone powder (DBP), tricalcium phosphate (TCP), or non-grafted controls. Pluronic polyols are easily mixed with either DBP or TCP to improve handling ease. Calvaria were harvested at 12 weeks postsurgery and evaluated histomorphometrically, by contact radiography with subsequent densitometric analysis, through energy spectrometry utilizing a scanning electron microscope, and by fluorescent microscopy. RESULTS: There was a significant difference in the percentage of bone fill among the control, TCP, and DBP only groups, P <0.05. The only significant difference within any of these groups was between the TCP control and TCP plus systemic F-127, P<0.05. CONCLUSIONS: Although there were isolated differences, the overall trend was that the pluronic polyol and the mode of administration did not result in a significant change in bone wound healing as measured by the percentage of bone fill. Pluronic polyols may be considered as carriers for osseous graft materials. PMID- 11895286 TI - Comparison of platelet-rich plasma, bovine porous bone mineral, and guided tissue regeneration versus platelet-rich plasma and bovine porous bone mineral in the treatment of intrabony defects: a reentry study. AB - BACKGROUND: A combination of platelet-rich plasma (PRP), bovine porous bone mineral (BPBM), and guided tissue regeneration (GTR) has been shown to be effective in promoting reduction in probing depth, gain in clinical attachment, and defect fill in intrabony periodontal lesions. The individual role played by PRP, BPBM, and GTR in this combined therapy is unclear and needs to be elucidated. The purpose of this study was to compare the clinical effectiveness of 2 regenerative techniques for intrabony defects in humans: a combination of PRP/BPBM/GTR versus a combination of PRP/BPBM. METHODS: Twenty-one patients participated in the study. Using a split-mouth design, interproximal bony defects were surgically treated with either a combination of PRP/BPBM/GTR or PRP/BPBM. The primary outcomes of the study included changes in probing depth, attachment level, and defect fill as revealed by reentry surgeries at 6 months post treatment. RESULTS: At 6 months postoperatively, clinical examination of the treated defects revealed that both treatment modalities resulted in significant probing depth reduction and clinical attachment gain compared to baseline values. Probing depth improvement was 3.98 +/- 1.02 mm on buccal and 3.94 +/- 0.94 mm on lingual sites for the PRP/BPBM group and 4.19 +/- 0.88 mm on buccal and 4.21 +/- 0.92 mm on lingual sites for the PRP/BPBM/GTR group. Gain in clinical attachment was 3.78 +/- 0.72 mm on buccal and 3.84 +/- 0.76 mm on lingual sites for the PRP/BPBM group and 4.12 +/- 0.78 mm on buccal and 4.16 +/- 0.83 mm on lingual sites for the PRP/BPBM/GTR group. Reentry surgeries revealed similar defect fill for both treatment groups (PRP/BPBM group: 4.82 +/- 1.34 mm on buccal and 4.74 +/ 1.30 mm on lingual sites; PRP/BPBM/GTR group: 4.96 +/- 1.28 mm on buccal and 4.78 +/- 1.32 mm on lingual sites). None of the differences between the 2 treatment groups was statistically significant. CONCLUSIONS: The results of this study show that both combinations of PRP/BPBM/GTR and PRP/BPBM are effective in the treatment of intrabony defects present in patients with advanced chronic periodontitis. The results also suggest that GTR adds no clinical benefit to PRP/BPBM. Further studies are necessary to assess the individual role played by PRP and BPBM in the clinical outcome achieved with their combination. PMID- 11895287 TI - Influence of nicotine administration on different implant surfaces: a histometric study in rabbits. AB - BACKGROUND: This study investigated the influence of implant surface on osseointegration around titanium implants inserted in the tibiae of rabbits administered with nicotine. METHODS: Thirty-two (32) New Zealand rabbits were included in the study. After anesthesia, the tibia surface was exposed and 2 screw-shaped commercially available pure titanium implants 7.0 mm in length and 3.75 mm in diameter were placed bilaterally. A total of 128 implants were inserted: 64 blasted with Al2O3 particles (Group 1) and 64 with a machined surface finish (Group 2). The animals were randomly assigned to 1 of 4 treatment subgroups, and daily subcutaneous injections of nicotine were administered: A) saline solution; B) 0.37 mg/kg; C) 0.57 mg/kg; and D) 0.93 mg/kg. In order to label regenerated bone, a 2% calcein green solution was administered by intramuscular injection at 0, 7, and 15 days after implant insertion. After 42 days, the animals were sacrificed and undecalcified sections were prepared. The degree of bone contact with the implant surface, the bone area, and the intensity of bone labeling were measured into the limits of the implant threads. RESULTS: Statistical analysis (2-way ANOVA) revealed no significant difference regarding the effect of nicotine on bone healing around the implants (P>0.05). However, a significant influence of the implant surface on the degree of bone-to-implant contact was detected in groups C (30.13 +/- 4.97 and 37.85 +/- 8.85, for machined and Al2O3-blasted surfaces, respectively) and D (27.79 +/- 3.93 and 33.13 +/- 8.87, for machined and Al2O3-blasted surfaces, respectively) (P<0.05). CONCLUSIONS: Although nicotine administration may not statistically influence bone healing around titanium implants, implant surface design may enhance osseointegration after nicotine administration. PMID- 11895288 TI - The effects of varying degrees of allograft decalcification on cultured porcine osteoclast cells. AB - BACKGROUND: Demineralized freeze-dried bone allograft (DFDBA) is widely used in periodontal therapy as a scaffold for new bone formation in periodontal defects. It is demineralized, theoretically, to expose osteoinductive or osteoconductive bone matrix proteins that should facilitate osteogenesis. The degree of DFDBA demineralization varies between tissue banks and may affect clinical regeneration. A 2% residual calcium level in DFDBA has been shown to result in the highest alkaline phosphatase activity levels in cultured human periosteal cells and is optimally osteoinductive or osteoconductive for new bone formation. The purpose of this study was to evaluate the effect of 4 different residual calcium levels in commercially available DFDBA samples on porcine osteoclast activity as measured by resorption on calcium phosphate-coated disks. METHODS: Bone marrow was harvested from the femurs of 3-week-old farm pigs and cultured for 3 weeks. Hematopoietic stem cells were allowed to differentiate into mature active polykaryons displaying genuine osteoclast characteristics. The osteoclast cells displayed a dense actin band inside the margins of the cytoplasm under light microscopy. Culture media was decanted and collagenase added to free the attached cells. Equal cell samples were pipetted onto calcium phosphate-coated disks in 24-well plates. DFDBA samples with 1.44%, 2.41%, and 5.29% residual calcium; FDBA (30% residual calcium); and control cultures without allograft samples were prepared and all samples incubated for 1 week. Cells were fixed and stained for tartrate-resistant acid phosphatase (TRAP), Oregon Green 488 phalloidin, a stain for cytoskeletal proteins, and counterstained with propidium iodide. Specimens were examined by light and fluorescence microscopy using epi illumination. Calcium phosphate disks were then rinsed in 5% sodium hypochlorite to remove adherent osteoclasts, and substrate surface changes were measured by white light interferometry and image analysis. RESULTS: A higher yield of TRAP positive cells was produced without DFDBA; however, resorptive activity appears to be significantly increased in the presence of 2.41% residual calcium as compared to all other experimental groups (P<0.0065). CONCLUSION: In this in vitro model, porcine osteoclasts show significantly more resorptive activity as measured on calcium phosphate-coated disks in the presence of 2.41% residual calcium in DFDBA than in other DFDBA residual calcium levels. PMID- 11895289 TI - Evaluation of frozen subepithelial connective tissue grafts to increase the zone of attached gingiva. Report of 5 cases. AB - BACKGROUND: In the past decade, gingival tissue has been regenerated using subepithelial connective tissue grafts (SECT). This report presents the use of frozen subepithelial connective tissue (FSECT) for increasing the zone of attached keratinized gingiva (KG). METHODS: Five cases, 4 females and 1 male, aged 55 to 73 years, presented with inadequate attached KG. The SECT grafts were harvested from the palatal flap during conventional flap surgery. All grafts were frozen immediately. Five weeks to 10 months later, the grafts were placed in areas in which patients had inadequate KG. RESULTS: In all cases, healing was uneventful. All cases were followed up 3 months to 1.5 years. An increase of 1 to 3 mm of KG was achieved. CONCLUSIONS: These cases show that the FSECT graft is useful in treating mucogingival problems. Further randomized controlled trials over long periods of time are necessary to establish whether this procedure offers long-term benefits to patients. PMID- 11895290 TI - A critical assessment of interleukin-1 (IL-1) genotyping when used in a genetic susceptibility test for severe chronic periodontitis. AB - BACKGROUND: This review addresses the ability of a commercially available genetic susceptibility test to determine the risk of developing severe chronic periodontitis. The test is used to detect the simultaneous occurrence of allele 2 at the IL-1A+4845 and IL-1B+3954 loci. If both of these polymorphisms are present, patients are referred to as being genotype-positive and considered predisposed to becoming afflicted with severe chronic periodontitis. A basic premise of this test is the assumption that individuals who are genotype-positive produce increased amounts of IL-beta in response to microbial lipopolysaccharides, which allegedly predisposes them to an exaggerated inflammatory response and an increased incidence of chronic periodontitis. METHODS: Controlled clinical trials were selected that evaluated the ability of the genetic test to predict which patients were susceptible to bleeding upon probing, periodontitis, peri-implantitis, and tooth loss. RESULTS: Comparison of results from test (genotype-positive) and control groups (genotype-negative) revealed that there is ambiguity with regard to predicting which patients will manifest elevated sub-gingival levels of IL-beta. Similarly, it is questionable if the test is able to forecast which individuals will demonstrate an increased occurrence of bleeding upon probing, diminished clinical attachment, decreased osseous support, or loss of teeth. CONCLUSIONS: There are many unanswered questions concerning the utility of detecting allele 2 at the IL-1A+4845 and IL IB+3954 loci to foretell which patients will develop severe chronic periodontitis. Therefore, clinicians must cautiously interpret results obtained with the commercially available genetic susceptibility test before they alter maintenance schedules or treatment regimens of symptomatic or asymptomatic patients. PMID- 11895291 TI - Siderophores of the human pathogenic fluorescent pseudomonads. AB - Bacteria need a sufficient supply of iron in ionic form for their metabolism. When living in an environment where this is not possible (as in the soil due to the presence of highly unsoluble ferric oxide hydrates, or in living organisms where iron is bound to peptidic chelators) Fe3+ complexing compounds, called siderophores, are produced. The siderophores of Pseudomonas aeruginosa, a dangerous opportunistic human pathogen, and of related potentially pathogenic species will be presented. PMID- 11895292 TI - Siderotyping--a powerful tool for the characterization of pyoverdines. AB - Tools for the identification of bacteria are of great importance especially for taxonomical and medical purposes. In the case of fluorescent pseudomonads a quick and unambiguous identification is possible by methods that are referred to as "siderotyping". All of them are based upon the characterization of the bacterial siderophores or the receptors expressed for the uptake of these compounds. Different microbiological and bioanalytical tests that are accurate, rapid and easy to use will be described. PMID- 11895293 TI - Multidrug efflux in Pseudomonas aeruginosa: components, mechanisms and clinical significance. AB - Pseudomonas aeruginosa is an opportunistic human pathogen characterized by an intrinsic resistance to multiple antimicrobial agents and the ability to develop high-level (acquired) multidrug resistance during antibiotic therapy. Much of this resistance is promoted by highly homologous three-component efflux systems of broad substrate specificity, of which four have been identified to date. These include MexA-Mexs-OprM and MexX-MexY-OprM, which are expressed constitutively in wild type cells and, thus, provide for intrinsic multidrug resistance, and MexC MexD-OprJ and MexE-MexF-OprN, whose expression so far has only been seen in acquired multidrug resistant mutant strains. Additional homologues of these efflux systems are identifiable in the recently released genome sequence, though their roles, if any, in antimicrobial efflux are unknown. These tripartite pumps are composed of an integral cytoplasmic membrane drug-proton antiporter of the resistance-nodulation-cell division (RND) family of exporters, a channel-forming outer membrane efflux protein (or outer membrane factor [OMF]) and a periplasmic membrane fusion protein (MFP) that links the other two. In addition to a number of antimicrobials of clinical significance, these pumps also export dyes, detergents, disinfectants, organic solvents and acylated homoserine lactones involved in quorum-sensing. While the natural functional of these pumps remains undefined, the fact that they contribute to antimicrobial resistance in P. aeruginosa makes them reasonable targets for therapeutic intervention. PMID- 11895294 TI - Structural biology of bacterial iron uptake systems. AB - Numerous bacterial proteins are involved in microbial iron uptake and transport and considerable variation has been found in the uptake schemes used by different bacterial species. However, whether extracting iron from host proteins such as transferrin, lactoferrin or hemoglobin or importing low molecular weight iron chelating compounds such as heme, citrate or siderophores, Gram-negative pathogenic bacteria typically employ a specific outer membrane receptor, a periplasmic binding protein and two inner membrane associated proteins: a transporter coupled with an ATP-hydrolyzing protein. Often, studies have shown that proteins with similar function but little amino acid sequence homology are structurally related. Elucidation of the structures of the Escherichia coli outer membrane siderophore transport proteins FepA and FhuA have provided the first insights into the conformational changes required for ligand transport through the bacterial outer membrane. The variations between the structures of the prototypical periplasmic ferric binding protein FbpA from Neisseria and Haemophilus influenzae and the unusual E coli periplasmic siderophore binding protein FhuD reveal that the different periplasmic ligand binding proteins exercise distinct mechanisms for ligand binding and release. The structure of the hemophore HasA from Serratia marcescens shows how heme may be extracted and utilized by the bacteria. Other biochemical evidence also shows that the proteins that provide energy for iron transport at the outer membrane, such as the TonB ExbB-ExbD system, are structurally very similar across bacterial species. Likewise, the iron-sensitive gene regulatory protein Fur is found in most bacteria. To date, no structural information is available for Fur, but the structure for the related protein DxtR has been determined. Together, these three dimensional structures complement our knowledge of iron transport systems from other pathogenic bacteria, including Pseudomonas aeruginosa, which has a number of homologous iron uptake proteins. More importantly, the current structures for iron transport proteins provide rational starting points for design of novel antimicrobial agents. PMID- 11895295 TI - Siderophore-antibiotic conjugates used as trojan horses against Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is a dangerous opportunistic bacterium responsible for frequently lethal hospital (nosocomial) infections. It endangers especially severely injured patients suffering from large wounds or severe burns, as well as persons whose immune system is weakened. An extremely critical situation exists for patients suffering from mucoviscidosis (cystic fibrosis), when P. aeruginosa infects the bronchial tubes. P. aeruginosa is resistant against many disinfecting agents and, more important, an increasing number of strains especially from hospital isolates have become highly resistant against most antibiotics. The low permeability of the outer membrane and an active export mechanism for low molecular weight substances are the main reasons for the resistance. In addition, beta-lactamase activity affects treatment with beta-lactam antibiotics. An approach to overcome the problem of resistance lies in the synthesis of antibiotics conjugated with compounds active as siderophores. In this way the transport ways for iron complexes into the cell can be used ("Trojan Horse strategy"), and the presence of large substituents reduces the export and the beta-lactamase activity. The results obtained with natural (pyoverdins) and synthetic (mainly catecholate) siderophores will be reviewed. PMID- 11895296 TI - Magnetic resonance imaging of patients with epilepsy. AB - Magnetic resonance imaging (MRI) is the radiological investigation of choice for the evaluation of patients with epilepsy. It is able to detect and characterize the structural origin of seizures, and significantly influences treatment planning and prognosis. The indications for MRI, protocols used for MRI in epilepsy and the relevant imaging anatomy are discussed. The major categories of epileptogenic lesions which result in chronic seizures are reviewed and illustrated. Mesial temporal sclerosis is emphasized, reflecting its major importance as a cause of medically intractable epilepsy. The role of MRI in the planning and assessment of epilepsy surgery is considered. PMID- 11895297 TI - Ultrasound of renal transplantation. AB - The most effective primary treatment of chronic renal failure is renal transplantation. A significant improvement in lifestyle and family life in conjunction with it being an extremely cost-effective procedure has resulted in an intense monitoring and imaging programme to help ensure a successful outcome. Ultrasound, both grey-scale and colour-flow Doppler, are useful monitoring techniques when interpreted in the clinical context, and in the delineation of peri-transplant collections, some of which can be drained under ultrasound guidance. After the early post-operative period it can also be utilized in the diagnosis of chronic vascular complications including transplant artery stenosis and arteriovenous fistula, although it is of limited use in the diagnosis of chronic rejection. This article will discuss the role of ultrasound in all its guises and how its efficacy in both the early transplant period in the monitoring of graft dysfunction and in the detection of the more chronic conditions including transplant artery stenosis and arteriovenous fistulae. A more limited role for ultrasound also exists in the long-term follow-up of patients and to aid the detection of complications including susceptibility to malignancy. PMID- 11895298 TI - Radiological features of epiploic appendagitis and segmental omental infarction. AB - Epiploic appendagitis and segmental omental infarction are more frequently encountered with the increased use of abdominal ultrasound and Computed tomography (CT) in the radiological assessment of the patient who presents clinically with acute abdominal pain. Recognition of specific imaging abnormalities enables the radiologist to make the correct diagnosis. This is important, as the appropriate management of both conditions is often conservative. Follow-up imaging features correlate with clinical improvement. PMID- 11895300 TI - A pilot evaluation of the R2 image checker system and users' response in the detection of interval breast cancers on previous screening films. AB - AIM: Breast screening radiologists are under considerable pressure to maximize cancer detection and minimize the number of interval cancers. This study used screening films, previously reported as normal, taken before the development of an interval cancer. The aim was to determine the sensitivity and specificity of the R2 Image Checker System and the responses of users in detecting abnormalities in these previous films. MATERIALS AND METHODS: The first R2 Image Checker System in the U.K. was installed in the Canterbury Assessment Centre for evaluation purposes. System performance was compared with the standard protocol for assessing interval cancers used in the NHS Breast Screening Programme (NHSBSP) Three readers examined the previous films blind before switching on R2 and compared the prompts generated with the location of the subsequent interval cancer. RESULTS: The previous screening films for 104 cases generated 134 R2 prompts on the subsequent cancer side and 109 on the non-cancer side. Readers classified 29 cancers as false-negative or minimal signs and R2 correctly prompted in 15 of these cases (52%). At least one reader rejected the correct prompt in five cases. CONCLUSION: From this preliminary study it appears that R2 overprompts normal areas and underprompts some cancers in these difficult cases. A full multi-centre evaluation is needed to assess the possible contribution of R2 to the NHSBSP. PMID- 11895299 TI - Predicting invasion in mammographically detected microcalcification. AB - AIM: To identify pre-operative factors which predict presence of invasive disease within mammographically detected malignant microcalcification. MATERIALS AND METHODS: A retrospective analysis was undertaken of 116 serial stereotactic core needle biopsies (SCNBs) performed on malignant mammographic calcification. Final surgical pathology was correlated with pre-operative features (clinical, radiological and core histology) in an attempt to predict the presence of an invasive component. RESULTS: Thirty-eight clusters contained invasive carcinoma. The sensitivity of SCNB for invasion was 55%. Clinical features, calcium morphology and cluster size were not shown to be predictive of invasive disease. Ductal carcinoma in situ (DCIS) of high grade on core histology and increasing number of calcifications were predictive of increased risk of invasion (high grade core biopsy DCIS and > 40 calcifications 48% invasive at surgical histology; high grade core biopsy DCIS and < 40 calcifications 15% invasive; non high grade core biopsy DCIS 0% invasive). CONCLUSIONS: Identification of those clusters diagnosed as DCIS by percutaneous biopsy which are likely to harbour an invasive component is possible. It would seem reasonable to consider staging the axilla at therapeutic surgery in these patients. PMID- 11895301 TI - Clinical validity of a normal pulmonary angiogram in patients with suspected pulmonary embolism--a critical review. AB - AIM: To determine the validity of a normal pulmonary angiogram in the exclusion of pulmonary embolism (PE), based on the safety of withholding anticoagulant therapy in patients with a normal pulmonary angiogram. MATERIALS AND METHODS: A review of English reports published between 1965 and April 1999 was carried out. Eligible articles described prospective studies in patients with suspected PE and a normal pulmonary angiogram, who remained untreated and were followed-up for a minimum of 3 months. Articles were evaluated by two authors, using pre-defined criteria for strength of design. End points consisted of fatal and non-fatal recurrent thromboembolic events. A sensitivity analysis was performed, by removing one study at a time from the overall results and by comparing pre- and post-1990 publications. RESULTS: Among 1050 patients in eight articles included in the analysis, recurrent thromboembolic events were described in 18 patients (1.7% 95% CI: 1.0-2.7%). These were fatal in three patients (0.3% 95% CI: 0.02 0.7%). The recurrence rate of PE decreased from 2.9% (95% CI: 1.4-6.8%) before 1990 to 1.1% (95% CI: 0.5-2.2%) after 1990. CONCLUSION: It would appear that the ability to exclude PE by angiography has improved over the years, as indicated by recurrence rate of PE. The low recurrence rate of PE supports the validity of a normal pulmonary angiogram for the exclusion of PE. PMID- 11895302 TI - Achilles tendon (TA) size and power Doppler ultrasound (PD) changes compared to MRI: a preliminary observational study. AB - AIM: To assess whether abnormal Achilles tendon (TA) magnetic resonance imaging (MRI) and spectral ultrasound (US) features have associated development of microvascular power Doppler (PD) flow. MATERIALS AND METHODS: In a prospective, controlled and blinded study six patients with TA symptoms were compared to five with other ankle abnormalities. Two radiologists independently measured the mean maximal anteroposterior diameter on MRI and conventional US (categorized as normal <6 mm, mild 6.1-10 mm, moderate 1.1-1.5 cm and severely enlarged > 1.6 cm), assessed morphology and studied the vessels using power Doppler. They formed a consensus over discrepancies. Sonography of the contralateral side within 24 h was used as a control. RESULTS: Twenty-one tendons in six women and five men, aged 45-77 years (mean 57.6 years), were examined, 12 tendons were of normal US morphology and size (< 6 mm), and did not exhibit PD's flow (interobserver agreement K > 0.74). Of the 12 tendons studied by MRI five were normal, seven tendons were enlarged, five of which had a proportionate increase in PD flow at the margin on the deep surface and four also had vessels in the centre of the tendon. All five of these tendons had high signal on T2-weighting (T2W). Of the two mildly enlarged tendons of intermediate signal on T1 and T2W, one showed PD flow and the other did not. CONCLUSIONS: In patients with TA disease power Doppler ultrasound shows proliferation of vessels in enlarged, abnormal tendons demonstrated on MRI and standard ultrasound, in the absence of definite tears. PMID- 11895303 TI - Apparent paradoxical vault changes with middle cranial fossa arachnoid cysts- implication for aetiology. AB - Three cases of middle cranial fossa arachnoid cyst with paradoxical bone changes in the adjacent vault are described, namely, a small middle cranial fossa and pneumosinus dilatans. This association is unusual and unique. The existing literature is reviewed and the probable aetiological factors discussed. PMID- 11895305 TI - Enterotubal fistulae secondary to tuberculosis: report of three cases and review of literature. PMID- 11895306 TI - Pulmonary arteriovenous malformations. PMID- 11895304 TI - The therapeutic barium enema revisited. PMID- 11895307 TI - Transcatheter treatment of profunda femoris aneurysm. PMID- 11895308 TI - Survey of intussusception reduction in England, Scotland and Wales: how and why we could do better. PMID- 11895309 TI - Re: Herron et al. Clin Radiol 2000; 55: 82-98. PMID- 11895310 TI - The public library of science and "open access" to the scientific literature. PMID- 11895311 TI - Bioavailability of selenium from raw or cured selenomethionine-enriched fillets of Atlantic salmon (Salmo salar) assessed in selenium-deficient rats. AB - The bioavailability of Se from raw and cured selenomethionine-enriched (Se enriched) salmon fillets was assessed in Se-deficient male albino rats (Mol: Wist). A low-Se Torula yeast feed was supplemented with 0, 50, 100, 150 or 200 microg Se/kg as sodium selenite or as Se from raw or cured Se-enriched salmon. The diets were fed to weanling rats for 10 and 30 d. Bioavailability of Se was assessed by metabolic balance, Se accumulation in femur, muscle, liver and plasma, and induction of Se-dependent glutathione peroxidase (EC 1.11.1.9; GSHpx) in plasma as response parameters. Except for the metabolic balance results, the slope-ratio method was used when calculating Se bioavailability from raw or cured Se-enriched fish fillets (test food) relative to sodium selenite (standard). The data for fractional apparent absorption and fractional retention showed differences (P<0.05) among all three Se sources in the order raw salmon > cured salmon > selenite. At 10 d, Se from raw and cured Se-enriched fish fillets tended to be more bioavailable than selenite. This was supported by the observations for Se accumulation in femur and muscle and induction of GSHpx activity. At 30 d, all response parameters showed a higher bioavailability of Se from raw and cured Se enriched fish fillets compared with selenite. Differences (P<0.05) in Se accumulation in muscle at 10 and 30d, and differences (P<0.05) in fractional apparent absorption and fractional retention suggested that curing salmon altered the utilisation of Se. The experimental results showed that enrichment of fish fillets with selenomethionine yields fillets with high Se bioavailability. PMID- 11895312 TI - Urinary excretion of catechin metabolites by human subjects after red wine consumption. AB - Little is known about flavonoid metabolism and excretion in man. In the present study, the urinary excretion of a major flavonoid in wine, catechin, and its metabolites, were measured after nine human subjects each consumed 120 ml red wine (RW) on one day and de-alcoholized red wine (DRW) on a separate day. Both the RW and DRW contained 120 (SEM 3) micromol catechin (35 mg). GC-MS analyses of the trimethylsilylated derivatives of catechin and 3' and 4' methylcatechin were performed before and after hydrolysis of conjugates by beta-glucuronidase and sulfatase. Baseline urine samples collected prior to wine consumption contained 0.013 (SEM 0.005) micromol catechin and metabolites. During the 8 h period following consumption of RW and DRW, 6.6 (SEM 0.9) and 5.3 (SEM 0.6) micromol catechin and metabolites were excreted in 893 (SEM 94) and 740 (SEM 101) ml urine respectively. This corresponded to 3.0-10.3% of the dose after RW and 2.1-8.2% of the dose after DRW. The amount of catechin and metabolites excreted in urine was 20% higher after RW compared with DRW (P=0.06). Catechin in all urine samples was present as metabolites and there were no differences in the proportions of individual metabolites after RW and DRW. As with other flavonoids, the fate of most ingested catechin is not yet known. PMID- 11895313 TI - Chronic oral administration of rhamnogalacturonan-II dimer, a pectic polysaccharide, failed to accelerate body lead detoxification after chronic lead exposure in rats. AB - Lead is a ubiquitous heavy metal and its toxicity remains an important public health issue. In previous work, we reported that ingestion of rhamnogalacturonan II dimer (dRGII), a pectic polysaccharide, may decrease intestinal absorption and status of Pb in rats. Here, we evaluated the potential detoxifying effect of different doses of dRGII after chronic oral Pb exposure in rats. For this purpose, six groups of ten male Wistar rats weighing 150g were treated as follows: group A received a semi-purified control diet for 6 weeks; groups B, C, D, E and F received the same diet plus 3 mg Pb (as acetate) for 3 weeks. Group B was then killed. Groups C, D, E, and F continued to receive the semi-purified control diet containing 0, 2, 6 or 18g dRGII/kg diet for 3 additional weeks. During the last 5 d, a Pb conventional balance study was performed. Rats were then anaesthetized and tissues were sampled for Pb and essential minerals assay. The results showed that residual Pb in the added dRGII was not available for absorption. However, the added dRGII failed to induce any significant increase in faecal or urinary Pb excretion. Consequently, at the end of the study the intestinal Pb absorption and balance remained unchanged in the animals receiving the different doses of dRGII. In line with this, we showed that dRGII administration was not effective in decreasing tibia or kidney Pb levels in rats. In conclusion, Pb complexed by dRGII in fruits and vegetables and fruit juice is thus mostly unavailable for intestinal absorption. However, the addition of dRGII after chronic Pb exposure does not help Pb detoxification. PMID- 11895314 TI - Dietary iodine intake and urinary iodine excretion in a Danish population: effect of geography, supplements and food choice. AB - I deficiency diseases remain a health problem even in some developed countries. Therefore, measurement of I intake and knowledge about food choice related to I intake is important. We examined I intake in 4649 randomly selected participants from two cities in Denmark (Copenhagen and Aalborg) with an expected difference in I intake. I intake was assessed both by a food frequency questionnaire and by measuring I in casual urine samples. I excretion was expressed as a concentration and as estimated 24-h l excretion. Further, subgroups with low I intake were recognized. I intake was lower in Aalborg than in Copenhagen for all expressions, and lower than recommended in both cities if I intake from supplements was not included. Milk was the most important I source, accounting for about 44% of the I intake, and milk (P<0.001) and fish (P=0.009) intake was related to I excretion in a multiple linear regression model. Thus, risk groups for low I intake were individuals with a low milk intake, those with a low intake of fish and milk, those not taking I supplements and those living in Aalborg where the I content in drinking water is lower. Even individuals who followed the advice regarding intake of 200-300 g fish/week and 0.5 litres milk/d had an intake below the recommended level if living in Aalborg. PMID- 11895315 TI - Correlates of plasma homocysteine, cysteine and cysteinyl-glycine in respondents in the British National Diet and Nutrition Survey of young people aged 4-18 years, and a comparison with the survey of people aged 65 years and over. AB - Plasma total homocysteine (tHcy), cysteine and cysteinyl-glycine were measured in a representative sample of 922 young people aged 4-18 years, participating in the National Diet and Nutrition Survey in mainland Britain in 1997. Both tHcy and cysteine increased markedly with age; cysteinyl-glycine less so. Neither tHcy nor cysteine differed between genders; cysteinyl-glycine was higher in males. tHcy concentrations were lowest in the winter; cysteine and cysteinyl-glycine varied only slightly with season. In respondents aged >15 years, tHcy was higher in smokers, but in respondents aged 7-11 years, tHcy was higher in those whose mothers smoked. tHcy was inversely correlated with serum folate, serum vitamin B12 and vitamin B6 status, but neither cysteine nor cysteinyl-glycine shared these relationships. The relationships between tHcy and B-vitamin status indices ran parallel with those of the 65 years and over survey, but at much lower tHcy concentrations for any given B-vitamin concentration. Age-adjusted tHcy was not correlated with anthropometric indices, blood pressure, haematology, plasma creatinine, urea or cholesterol, but was directly correlated with fasting triacylglycerol. We conclude that disease-risk indices, like tHcy and perhaps cysteine, if established during early life, may be modulated by diet and lifestyle, thereby providing an opportunity for public health intervention. PMID- 11895316 TI - Nutritional status, brain development and scholastic achievement of Chilean high school graduates from high and low intellectual quotient and socio-economic status. AB - The objective of the present study was to investigate the inter-relationships between nutritional status (past and current nutrition), brain development, and scholastic achievement (SA) of Chilean high-school graduates from high and low intellectual quotient (IQ) and socio-economic status (SES) (mean age 18.0 (SD 0.9) years). Results showed that independently of SES, high-school graduates with similar IQ have similar nutritional, brain development and SA variables. Multiple regression analysis between child IQ (dependent variable) and age, sex, SES, brain volume (BV), undernutrition during the first year of life, paternal and maternal IQ (independent variables) revealed that maternal IQ (P<0.0001), BV (P<00387) and severe undernutrition during the first year of life (P<0.0486), were the independent variables with the greatest explanatory power for child IQ variance (r2 0.707), without interaction with age, sex or SES. Child IQ (P<0.0001) was the only independent variable that explained both SA variance (r2 0.848) and academic aptitude test variance (r2 0.876) without interaction with age, sex or SES. These results confirm the hypotheses formulated for this study that: (1) independently of SES, high-school graduates with similar IQ have similar variables of nutritional status, brain development and SA; (2) past nutritional status, brain development, child IQ and SA are strongly and significantly inter-related. These findings are relevant in explaining the complex interactions between variables that affect IQ and SA and can be useful for nutritional and educational planning. PMID- 11895317 TI - Evidence in support of a concept of reductive stress. PMID- 11895318 TI - Food choice ideologies: the modern manifestations of normative and humanist views of the world. AB - Two studies examined whether everyday food choice motives (FCMs) and abstract values constitute food choice ideologies (FCIs), whether these ideologies reflect the same normativism-humanism polarity as Tomkins' theory suggests to reflect ideologies in general, and whether various dietary groups endorse FCIs in different ways. In Study 1, 82 female participants filled in the Food Choice Questionnaire, a short version of Schwartz's Value Survey, and Tomkins' Polarity Scale. The results reflected four FCIs: ecological ideology (EI), health ideology (HI), pleasure ideology (PI) and convenience ideology (CI). Study 2 (N=144) replicated the results for ecological and health ideologies but not for pleasure and convenience ideologies. In both studies, EI, which was typical for vegetarians, was associated with a humanist view of the world, whereas HI was related to a normative view of the world. The results suggest that food choice has become a new site where one expresses one's philosophy of life. PMID- 11895319 TI - Effect of sucrose and safflower oil preloads on short term appetite and food intake of young men. AB - The effects of carbohydrate and fat on satiety have been examined primarily through meal composition studies. The purpose of this study was to compare the effects of pure sucrose and safflower oil, isovolumetric beverage preloads, on appetite (measured every 15 minutes by visual analogue scales) and food intake 60 minutes later. Young men consumed 0, 418, 836 and 1254 kJ of sucrose in the first two experiments and these same doses of safflower oil in the third. Finally, the largest doses of sucrose and safflower oil were compared. Sucrose, but not safflower oil, suppressed average appetite compared with control. In experiment 2, food intake was reduced (p<0.05) by 518 kJ after the 418 and 836 kJ preloads and by 1129 kJ after the 1254 kJ sucrose preload. Only the 1254 kJ dose of safflower oil significantly suppressed food intake by 480 kJ in the third experiment. When the 1254 kJ doses were compared directly, sucrose suppressed food intake by 653 kJ compared with control where as safflower oil did not. It is concluded that, in the short-term, sucrose produces a dose dependent reduction in appetite and food intake that is greater than that produced by safflower oil. PMID- 11895320 TI - Effects of dietary restraint on flavour-flavour learning. AB - Flavour preference learning in 21 restrained and 21 unrestrained females was explored using an evaluative conditioning paradigm. Each participant was exposed to an adapted version of the procedure used by Johnsrude et al., (1999, Learning & Motivation30, 250-264). During conditioning, participants sampled 10 instances each of three novel flavours presented in a semi-randomized order. After sampling a flavour they were instructed to eat a sweet according to three different flavour-reinforcement contingencies. One flavour was accompanied by a sweet on 90% of trials and was presented alone on 10% of the trials, while the remaining flavours were rewarded at ratios 50% : 50% and 10% : 90%, respectively. The conditioning phase occurred in conjunction with a counting task requiring continuous use of working memory, and was immediately followed by the participants making hedonic ratings of each flavour. Very few participants showed awareness of the purpose of the experiment or the specific reward contingencies. Despite this, the ratings given by the unrestrained eaters were highly correlated with the reward ratio experienced during conditioning. In contrast, restrained eaters exhibited no evidence for evaluative learning. These findings may explain the equivocal nature of results from previous studies of positive flavour-flavour learning (FFL) and may offer a novel theoretical context within which to study dietary restraint. PMID- 11895321 TI - Body image and weight consciousness among South Asian, Italian and general population women in Britain. AB - Italians in Britain have low rates of coronary heart disease while South Asians have high rates, which correspond to a tendency to central abdominal fat deposition and overweight. World variations in attitudes to body size are thought to be related to economic security. This cross-sectional study employed a range of measures including photographic silhouettes of known BMI to investigate the attitudes of 259 South Asian, Italian and general population women (aged 20-42 years) towards body size. Migrants are compared with British-born minority members. Our results indicate that although migrant South Asians were less happy with their weight than migrant Italians, fewer had tried to lose weight in the past or had experienced external pressures to change their bodies. More migrant South Asians than Italians or general population women equated one of the four largest shapes (BMI 28-38) with health and successful reproduction. All groups wanted to resemble one of the two thinnest shapes, equating them with longevity, likelihood of marriage and job success. British-born South Asians generally showed a considerable degree of convergence towards general population women's negative attitudes to large body size, but British-born Italians' attitudes were significantly more negative even than general population women. The study's conclusions were that South Asian health beliefs are an important focus of resistance to slimness. The tendency of migrant South Asians to equate large size with health contrasts with the opposing views of Italian and general population women. British-born South Asians' views are modifying from those of migrants, but significant differences remain when compared with general population women and British-born Italians. Present differences in economic security offer only a partial explanation; South Asian attitudes may be explained by economic insecurity in the past. PMID- 11895322 TI - Factor-analytic structure of food preferences in four-year-old children in the UK. AB - Studying the correlative structure of young children's food preferences may help us to understand the influences on the development of eating habits. We investigated patterns of food preferences in 214 same-sex twin pairs aged 4 to 5 years. Mothers completed a questionnaire of their children's likes and dislikes for 94 common foods. The proportion of children having tried each food, and their liking for it, were examined. These two measures were strongly positively correlated (r=0.61). Principal components analysis was used to examine the inter relationships among preferences for 76 foods that had been tried by at least 75% of the children. Four factors emerged, interpreted from food factor loadings as "Vegetables", "Desserts", "Meat and Fish", and "Fruit", explaining 24% of the variance. Thus, children tended consistently to like or dislike foods within each of these factor categories, independently of patterns of liking in the other factors. Factor loadings were unrelated to average liking for or likelihood of exposure to these foods. The distribution of factor loadings refutes the common assumption that simple sensory properties such as sweetness, saltiness or fattiness predict food preferences. Instead, a preference structure may develop for more complex multimodal sensory syntheses, which could have both biological and learnt bases. Future work comparing the food preference structure of mono- and dizygotic twins should help determine the relative contribution of genetic and environmental influences. PMID- 11895323 TI - Body weight, body-weight concerns and eating styles in habitual heavy users and non-users of artificially sweetened beverages. AB - This study investigated reported body weight, concerns about body weight and eating styles in habitual heavy users (consume>825 ml/day) and habitual non-users of artificially sweetened beverages (ASBs). Groups of habitual heavy users (N=51) and non-users (N=69) were compared on measures of weight using self-reported body mass index (BMI), and measures of weight concern and eating style using the Dutch Eating Behaviors Questionnaire (DEBQ), the Yale Eating Patterns Questionnaire (YEPQ), and the Eating Disorder Inventory (EDI). Habitual heavy users reported higher body weights (BMI), greater concerns about weight and greater tendencies toward certain eating styles, when compared to non-users. Using logistic regression, 82% of respondents were correctly classified as heavy or non-users of ASBs using body mass index and body-weight concerns. Associations between a heavy use of ASBs and certain eating styles can be explained by association with high body weights and concerns about body weight. PMID- 11895324 TI - Predictors of maternal child-feeding style: maternal and child characteristics. AB - We investigated relationships among maternal and child characteristics, and two aspects of maternal child-feeding styles that may place daughters at risk for developing problems with energy balance. Participants included 104 overweight (BMI> or =25) and 92 non-overweight (BMI<25) mothers and their 5-year-old, non Hispanic, White daughters. Child-feeding styles included (a) restriction of daughters' intake of energy-dense snack food, and (b) pressure to eat more food. Predictors of child-feeding styles included measures of (1) maternal investment in weight and eating issues, including dietary restraint and weight concern, (2) child adiposity, (3) maternal perceptions of the child as underweight or overweight, and (4) maternal concern for child weight. Mothers reported using more restrictive feeding practices when they were invested in weight and eating issues, when they perceived daughters as overweight, when they were concerned about daughters' weight, and when daughters were heavier. Mothers reported using more pressure in child feeding when daughters were thinner, and when mothers perceived daughters as underweight. Further analyses examined whether relationships among child-feeding styles were different for overweight and non overweight mothers. Overweight mothers' child-feeding styles appeared to be influenced by observable child weight characteristics, concerns for the child's weight status, and mothers' own history of overweight. Non-overweight mothers' child-feeding styles appeared to be influenced by distorted perceptions of and concerns for children, as well as distorted self-perceptions. PMID- 11895325 TI - Stability of body image and body image dissatisfaction in American college students over about the last 15 years. AB - It is widely assumed that body image dissatisfaction is increasing, particularly in females. We examined data from comparable samples, University of Pennsylvania introductory psychology students, over a span of about 15 years (1983-1984 versus 1995-1998). Ratings of current and ideal body figure were obtained using silhouettes, along with self-reported height and weight. While males always had a much smaller discrepancy between current and ideal than females, levels of dissatisfaction and gender differences in satisfaction have remained the same in these samples. This finding contrasts with the conclusion of a meta-analysis by Feingold and Mazzella in 1998 (Psychological Science 9 (3), 190-195), which indicates an increased difference in body image satisfaction between men and women over the last two decades. Possible accounts for this difference in results are discussed. PMID- 11895326 TI - High fibre breakfast cereals reduce fatigue. PMID- 11895327 TI - Experimental comparison of different techniques to measure saliva. PMID- 11895328 TI - Paradoxical effects of a high sucrose diet: high energy intake and reduced body weight gain. PMID- 11895329 TI - Water ingestion improves subjective alertness, but has no effect on cognitive performance in dehydrated healthy young volunteers. PMID- 11895331 TI - The activation of the adipose tissue associated with lymph nodes during the early stages of an immune response. AB - The effects of repeated local immune challenges with lipopolysaccharide (LPS) over 24 h on basal and noradrenaline-stimulated lipolysis and the development of sensitivity to interleukin-4 and tumour necrosis factor-alpha in adipocytes associated with lymph nodes were studied in adult guinea-pigs. Properties characteristic of perinodal adipocytes appeared in adipocytes at least 10 mm from the locally stimulated popliteal lymph node within 12 h, and in other node containing depots over 24 h. All effects appeared first in perinodal adipocytes and spread as though in response to signals emanating from the enclosed lymph node. The popliteal depot was more completely activated than the mesenteric, but its maximum rate of lipolysis/100 adipocytes was lower. None of the pre treatments in vivo, nor incubation with cytokines in vitro modulated lipolysis in adipocytes from the nodeless perirenal depot. The sensitivity of the perinodal adipocytes to cytokines changed within 3 h of immune stimulation, preceding detectable increases in lipolysis. Cytokine-stimulated and noradrenaline stimulated lipolysis sum, suggesting separate pathways. We conclude that sustained local activation of a single popliteal lymph node recruits additional adipocytes in node-containing depots only. Signals spread from lymph nodes to surrounding adipocytes, but the time courses of activation of adipocytes and their maximum responses differ between the mesenteric and popliteal depots. PMID- 11895330 TI - Comparison of the functional properties of murine dendritic cells generated in vivo with Flt3 ligand, GM-CSF and Flt3 ligand plus GM-SCF. AB - Flt3 ligand (FL) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are important growth factors for dendritic cells (DC). Substantial numbers of DC can be generated in vivo following the administration of either factor. We sought to extend our knowledge of the functional properties of these cells including their ability to prime naive CD8(+) T cells. In addition, we compared the nature of the DC generated in vivo with the single cytokines to those generated with the combination of FL+polyethylene glycol-modified GM-CSF (pGM-CSF). Treatment with FL+pGM-CSF yielded greater numbers of both CD11b(low) and CD11b(high) DC than with either cytokine alone, and these DC were more efficient at antigen (Ag) capture. The FL+pGM-CSF-generated CD11b(low) DC lacked expression of CD8alpha. Following treatment with LPS in vivo, all DC subsets upregulated CD40, CD80, CD86, and MHC class II expression, but surprisingly Ag capture was not downregulated and some DC subsets retained expression of intracellular MHC class II vesicles. Thus, even after activation in vivo with LPS, DC retained Ag capture properties of immature DC, and Ag presentation/costimulation properties of mature DC. Though all DC subsets stimulated CD4(+) T cell proliferation equivalently, FL generated DC were more efficient at priming Ag-specific CD8(+) cytolytic T cells than DC generated with either pGM-CSF alone or FL+pGM-CSF, and CD11b(high) DC were more efficient at priming CD8(+) T cells than CD11b(low) DC. PMID- 11895332 TI - Demonstration of biological activity of CD40 ligand (CD154) in the domestic cat. AB - The interaction between CD40L (CD154) on T cells and CD40 on antigen-presenting cells induces expression of accessory molecules that facilitate immune activation. Therefore, CD40L may have utility as an adjuvant for the development of potent antigen-specific immune responses following vaccination. As there was no information about the feline homologue of CD40L or its function, we generated stable cell lines expressing cDNAs encoding the feline CD40L homologue. As a preliminary to investigating the use of CD40L as an adjuvant for vaccination of the domestic cat, we tested the biological activity of the feline cytokine molecule in vitro. We demonstrated that cells expressing feline CD40L induced proliferation of feline peripheral blood mononuclear cells (PBMC) and that purified B cells could be induced to proliferate in response to feline CD40L. PMID- 11895333 TI - The effects of IL-6 and IL-10 and their specific antibodies in the acute inflammatory responses induced by carrageenan in the mouse model of pleurisy. AB - This study evaluates the effects of intrapleural (i.pl.) injection of interleukin (IL-6) and IL-10 and their specific antibodies on the early (4 h) and late (48 h) inflammatory responses caused by carrageenan (Cg) injected into the mouse pleural cavity. The i.pl. injection of IL-6, 5 min prior to Cg, reduced in a dose dependent and significant manner, the exudation and total and differential leukocyte migration according to assessment in both the early (4 h) and the late (48 h) phases of Cg inflammatory response (P<0.01). Intrapleural injection of IL 10, 5 min prior to i.pl. injection of Cg, resulted in a significant inhibition of the early phase (4 h) (P<0.01), but had no significant effect in relation to the late (48 h) phase of Cg response. The antibodies anti-IL-6 (given i.pl. 30 min prior to Cg) caused a significant decrease in both total and differential leukocyte influx, but significantly increased exudation according to assessment 4 h after pleurisy induction by Cg (P<0.01). In contrast, anti-IL-10 antibody caused graded and marked increase of both total and differential leukocyte influx and also increased fluid leakage as assessed 4 h after Cg injection (P<0.01). In the late phase (48 h) these antibodies increased the inflammatory parameters (anti-IL-6) studied or had no effect (anti-IL-10). Taken together, the current results confirm and extend previous data from the literature by showing that IL-6 and IL-10 regulate several signs of inflammatory response, here characterized by marked inhibition of polymorphonuclear cell influx and blockage of fluid leakage to the site of Cg-induced pleurisy in the mouse. PMID- 11895334 TI - The healing skin wound: a novel site of action of the chemokine C10. AB - To gain insight into the molecular mechanisms underlying the wound repair process, we searched for genes that are regulated by skin injury. For this purpose we generated a subtractive cDNA library from normal mouse back skin and 1 day full-thickness excisional wounds. One of the differentially expressed genes encodes the chemokine C10. Using Northern blotting, RNase protection assay and Western blotting, we confirmed the injury-induced expression of C10 at the mRNA and protein level. Maximal levels of C10 mRNA and protein were seen at day 1 after wounding, and expression levels subsequently declined. In situ hybridization and immunohistochemistry revealed expression of C10 in macrophages of the clot and the granulation tissue as well as in keratinocytes of the epidermis and the hair follicles at the wound edge. Since C10 is a potent chemoattractant for macrophages, our results suggest that this chemokine contributes to the strong macrophage influx observed in the healing skin wound. PMID- 11895335 TI - Possible correlation between high levels of IL-18 in the cord blood of pre-term infants and neonatal development of periventricular leukomalacia and cerebral palsy. AB - Periventricular leukomalacia (PVL) is a neonatal white matter damage of the brain of pre-term infants that often leads to cerebral palsy (CP). At present, diagnosis of PVL can be done by magnetic resonance imaging (MRI) and ultrasonography only when the infant is at least one week of age. No biochemical methods are available to identify high-risk infants at birth. Cytokines are usually not present in the cord blood but recently an elevation of IL-6 and TNF alpha levels has been reported in amniotic fluid, cord blood and brain sections of infants with white matter damages. Levels of interleukin-18 (IL-18), a pleiotropic cytokines expressed in the brain and many other tissues, are highly sensitive to pathophysiological changes to raise the possibility that IL-18 may provide a useful indicator of PVL. The cord blood from 17 pre-term infants with PVL, 38 pre-term infants without PVL, and 30 normal full-term infants were retrospectively analysed for IL-18, IL-6, IL-1beta, and TNF-alpha. The possible factors involved in alteration of IL-18 concentration in relation to PVL and CP were examined. IL-18 is undetectable in the cord blood of normal full-term infants. However, high levels of IL-18 exist in the cord blood samples obtained from pre-term infants who neonatally developed PVL followed by CP. For pre-term infants under 35 weeks of gestation, seven out of eight showing more than 200 pg/ml of IL-18 (87.5%) developed PVL neonatally, with five of them subsequently developing CP. In contrast, only five out of 38 pre-term infants with less than 100 pg/ml of IL-18 (13.2%) developed PVL. For pre-term infants with less than 30 weeks of gestation, eight out of nine showing more than 100 pg/ml of IL-18 (88.9%) developed PVL, with six of these eight (75%) developing CP later. In conclusion, the presence of high levels of IL-18 in the cord blood of the pre term infants is correlated with the incidence of PVL and CP and may provide a prognostic marker applicable at birth. PMID- 11895337 TI - Obstetric brachial plexus palsy. PMID- 11895338 TI - Obstetrical brachial plexus palsy. Prediction of outcome in upper root injuries. AB - Thirty obstetrical brachial plexus palsies involving the upper roots were retrospectively reviewed. There were 20 C5-C6 palsies and ten C5-C6-C7 palsies in which recovery of C7 occurred by the end of the first month. Recovery of elbow flexion at 3 months, C7 involvement and high birthweight were the best early predictors of outcome, but all were unreliable when used separately. In combination, recovery of elbow flexion and birthweight predicted the final outcome reasonably satisfactorily, particularly when elbow flexion at 9 months, and not 3 months was considered (risk of error = 13%). Brachial plexus reconstruction may therefore be justified when there was initial C7 involvement associated with increased birthweight and poor elbow flexion at 6-9 months. PMID- 11895339 TI - Assessment of the method and timing of repair of a brachial plexus traction injury in an animal model for obstetric brachial plexus palsy. AB - A Sunderland type IV traction injury to the C6 root of adult sheep or newborn lamb brachial plexus was used as a model for obstetric traction injury to the C5 root in humans. In one experimental cohort the injury was created and repaired using interfascicular nerve autografts or coaxially aligned freeze-thawed skeletal muscle autografts in a group of adult sheep and in a group of newborn lambs. In a second cohort a similar injury was created and repaired either immediately or after a delay of 30 days, using either interfascicular nerve autografts or coaxially aligned freeze-thawed skeletal muscle autografts in four groups of six newborn lambs. In all cases both functional and morphometric indices of nerve regeneration were poorer in the injured and repaired nerves than in normal nerves. In lambs the method of repair made no difference and no significant differences were found for any of the indices of nerve function or morphology. In sheep the use of muscle grafts was associated with a poorer outcome than the use of nerve autografts. Where a delay of 30 days had elapsed between injury and repair, the results using nerve autografts were not significantly different. Where freeze-thawed muscle autografts had been used, the maturation of the regenerated nerve fibres after delay was significantly poorer than after immediate repair. The electrophysiological variables CV(max) and jitter, which may be applied clinically, were found to be good discriminators of recovery in all of the animals and in respect of all procedures. PMID- 11895340 TI - Surgical correction of supination deformity in children with obstetric brachial plexus palsy. AB - We present a series of 40 children who were operated on for supination contracture following severe obstetric brachial plexus palsy. Surgery was done at an average age of 7 years and the mean postoperative follow-up was 4 years. In the 23 cases treated by an open or closed radial osteotomy, the mean intraoperative derotation was 78 degrees, the immediate postoperative position was 29 degrees pronation and it stabilized at follow-up at 17 degrees pronation. Biceps rerouting was performed in 17 cases without any recurrence of supination deformity and the final position was 22 degrees pronation. Some active forearm rotation was obtained in a few cases. These surgical corrections are part of an overall treatment plan and allow the "begging hand" to be corrected to a more functional and less noticeable position. PMID- 11895341 TI - Juxta-epiphyseal fractures of the base of the proximal phalanx of the fingers in children and adolescents. AB - A series of 34 juxta-epiphyseal fractures of the base of the proximal phalanx of the fingers of children and adolescents are presented. The pattern of injury appeared identical in all these fractures, with a lateral angulation force separating a small triangular metaphyseal fragment from the base of the phalanx on the side of angulation and the fracture line then continuing through the metaphysis, 1-2 mm distal to the growth plate. Fractures were classified into two types according to the degree of displacement. Type 1 fractures (n=18) were mildly displaced and were all successfully treated with closed reduction and splinting. Type 2 fractures (n=16) were severely displaced and problems with obtaining an adequate reduction and long-term residual deformities were encountered. One patient with a severely displaced fracture required open reduction and Kirschner-wire fixation because of flexor tendon entrapment at the fracture site. Another five cases required Kirschner-wire fixation after closed manipulation in order to maintain the reduction. The remaining 10 patients with Type 2 fractures were treated by closed reduction and splinting, and two patients healed with malunion causing a "pseudo-claw" deformity. PMID- 11895342 TI - Condylar advancement osteotomy for correcting condylar malunion of the finger. AB - Unicondylar fractures of proximal and middle phalanges of the finger can unite with intra-articular malunion, which may result in joint pain, stiffness and deformity. There is currently no satisfactory technique of corrective osteotomy for these fractures. Extra-articular osteotomies often do not give good results and existing techniques of intra-articular osteotomy through the healed fracture site are technically difficult due to the small bone fragment, difficulty achieving stable fixation and the risk of avascular necrosis. We propose a different method of intra-articular correction with a longitudinal osteotomy and advancement of the malunited condyle. The "condylar advancement osteotomy" can overcome problems encountered with the other techniques. Excellent results were obtained in six patients. PMID- 11895344 TI - Percutaneous cannulated screw fixation of acute scaphoid waist fracture. AB - This prospective study assessed the outcome of percutaneous cannulated screw fixation in 49 of 60 acute scaphoid fractures. The union rate was 100% (mean time for radiological union at 12 weeks). There were no early or mid-term complications and all achieved an excellent functional recovery. PMID- 11895345 TI - Genetic susceptibility in Dupuytren's disease: lack of association of a novel transforming growth factor beta(2) polymorphism in Dupuytren's disease. AB - The genes involved in the pathogenesis of Dupuytren's disease have yet to be identified. In this study, we tested for an association between Dupuytren's disease (DD) and a novel insertion polymorphism within the 5'-untranslated region (5'-UTR), of the TGFbeta(2) gene. DNA samples from 179 DD patients and 187 ethnically matched controls were examined. There was no statistically significant difference in TGFbeta(2) allele frequency distributions between cases and controls for the TGFbeta(2) polymorphism. PMID- 11895343 TI - Unstable scaphoid fracture nonunion: a medium-term study of anterior wedge grafting procedures. AB - A retrospective review of 37 patients with scaphoid fracture nonunions treated by interpositional bone grafting and internal fixation was conducted at an average follow-up of 5.7 years. Solid radiographic union was achieved in 35 cases. Preexisting avascular necrosis was a major adverse factor for achievement of union and satisfactory outcome. Based on the modified Mayo wrist-scoring system, 15 patients had an excellent result, 11 had a good result, four had a fair result and seven had a poor result. Patients with preexisting degenerative changes had a significantly worse clinical outcome. The vast majority of the patients had satisfactory correction of scaphoid length and the associated dorsal intercalated segment instability (DISI). Although 30 patients showed radiographic evidence of mild or moderate degenerative changes at their latest follow-up, there was no significant progression of arthrosis and the scaphoid nonunion advanced carpal collapse deformity did not progress after healing of the fracture nonunion. PMID- 11895346 TI - Dupuytren's contracture and sarcoma. AB - In order to study possible connections between Dupuytren's contracture and sarcoma we analysed the records of 18 patients who developed sarcoma 5 years or more after surgery for Dupuytren's contracture. We found an increased frequency of fibrosarcoma and malignant fibrous histiocytoma, but these patients did not differ from the other patients in the study group. Our analysis suggests that neither smoking, diabetes nor cancer syndromes can explain why patients with Dupuytren's contracture have a higher incidence of sarcoma. PMID- 11895347 TI - Comparative study of "staples versus sutures" in skin closure following Dupuytren's surgery. AB - This prospective, randomized controlled trial assessed the use of staples for closure of the palmar skin following Dupuytren's surgery. Although staples were significantly quicker to insert than sutures, patients experienced significantly more pain on removal of staples. There was no difference in the cosmetic appearance of the wound in the two groups. We recommend use of staples for palmar wound closure following long procedures. PMID- 11895348 TI - Outcome after corrective osteotomy for malunited fractures of the distal end of the radius. AB - This retrospective study evaluated the outcome of corrective osteotomy for malunited distal radial fractures and investigated the influence of the radiological result on the clinical outcome. Twenty-nine patients underwent corrective osteotomy for malunited, dorsally tilted fractures of the distal radius and 20 underwent corrective osteotomy for malunited, palmarly angulated distal radial fractures. All were surveyed at an average of 18 months after surgery and assessed for: pain; grip strength; range of motion; radial tilt; radial inclination; and ulnar variance. Postoperative radial tilt, radial inclination and ulnar variance were significantly improved by corrective osteotomy. Patients with no, or only minor residual deformity after corrective osteotomy had significantly better results than those with gross residual deformity. PMID- 11895349 TI - Radiolunate arthrodesis in the rheumatoid wrist: a retrospective clinical and radiological longterm follow-up. AB - A retrospective study was performed to investigate the clinical and radiological results of radiolunate arthrodesis in the rheumatoid wrist. Ninety-one wrists in 78 patients were assessed at a mean follow-up of 60 months. Most patients were pain-free and content with the overall result. In 68 wrists the carpus had been repositioned or maintained in neutral or slightly ulnar alignment and no further translation occurred. Midcarpal dislocation occured in ten and midcarpal rotation in 13 wrists. The midcarpal joint underwent further arthritic destruction in 34 wrists and secondary arthrosis in 32 wrists. In 25 wrists the midcarpal joint space remained unchanged. Radiolunate arthrodesis can successfully be performed in wrists even with advanced destruction. In cases with fixed carpal collapse, anatomical repositioning of the lunate and restoration of carpal height should not be attempted as this causes midcarpal dislocation or rotation or precipitates secondary arthrosis. PMID- 11895350 TI - The arterial system of the first intermetatarsal space and its influence in toe to-hand transfer: a report of 53 long-pedicle transfers. AB - A study of the arterial system in the first intermetatarsal space was performed during 53 toe-to-hand or vascularized tissue of the toe transfers with long arterial pedicles. The first dorsal metatarsal artery was used as a single pedicle in 25 transfers and the first plantar metatarsal artery in 21: both arteries were taken simultaneously in seven transfers. In 11 transfers, the dorsalis pedis artery and the perforating branch were absent, although this did not exclude the presence of a first dorsal metatarsal artery which arose from the plantar system by means of an ascending vascular branch in seven cases. PMID- 11895352 TI - Nerve conduction studies for carpal tunnel syndrome: essential prelude to surgery or unnecessary luxury? AB - Although carpal tunnel syndrome is a relatively trivial condition, controversy surrounds the use of nerve conduction studies, and whether they are essential to make the diagnosis, or as a prelude to surgery. This is partly due to the lack of a generally agreed definition of the condition, and failure to recognize that the patient's first priority is rapid relief of symptoms. If nerve conduction studies do not contribute to achieving that aim it would be better not to do them. Supporters of routine preoperative nerve conduction studies ignore their shortcomings, which include lack of standardization, absence of population-based reference intervals, and lack of sensitivity and specificity. Only a controlled trial, in which patients are randomized to receive treatment either with or without nerve conduction studies, will determine whether they improve the outcome in patients with a firm clinical diagnosis of carpal tunnel syndrome. PMID- 11895351 TI - Upper extremity infections following common carp fish (cyprinus carpio) handling. AB - Upper extremity infection caused by aquatic pathogens on fish is a well recognized clinical entity. We report five consecutive cases of upper extremity infections, ranging from a simple localized reaction to a life-threatening systemic illness, which developed after handling common carp fish (Cyprinus carpio). In four cases, infection occurred following a penetrating injury by either the bones or the fin spines of the fish. Vibrio vulnificus was isolated from wound aspirates in four cases. Early broad-spectrum antibiotic therapy is mandatory. Deterioration in the clinical condition or a poor response to conservative treatment requires a meticulous surgical drainage and excision of both infected and necrotic tissues. PMID- 11895353 TI - The carpal detachment injury of the triangular fibrocartilage complex. AB - Triangular fibrocartilage complex (TFCC) injuries were suspected clinically in 22 wrists of 21 patients, but arthrography and MRI assessments of this structure were normal. As conservative therapy for 2 months did not improve their symptoms, wrist arthroscopy was then performed. Although no abnormalities of the TFCC and ligaments were observed, meniscus homologue-like tissue which arose from TFCC was riding on the articular surface of the triquetrum. After resection of this soft tissue with a shaver and a punch, the symptoms disappeared in all cases. The arthroscopic findings suggested that a portion of TFCC that was originally attached to the ulnar side of the triquetrum had become detached. PMID- 11895354 TI - Treatment of soft tissue injuries to the dorsum of the metacarpophalangeal joint (Boxer's knuckle). AB - We retrospectively reviewed the surgical treatment for 16 cases of traumatic soft tissue injury to the metacarpophalangeal joint (Boxer's knuckle). A history of trauma was present in all cases and there was an associated extensor tendon dislocation in seven cases. Eight cases were initially treated conservatively, but their symptoms persisted. Intraoperative findings included rupture of the extensor hood or joint capsule in all cases. Surgical closure of the rupture of the joint capsule resulted in a successful outcome in all cases. We consider that conservative treatment of this injury may not be effective when the joint capsule of the metacarpophalangeal joint is ruptured. We recommend arthrography of the metacarpophalangeal joint to assist in the decision as to whether to proceed with surgical or conservative treatment. PMID- 11895355 TI - Functional outcome following salvage of failed trapeziometacarpal joint arthroplasty. AB - The outcomes in 12 patients who underwent revision surgery for a failed trapeziometacarpal joint arthroplasty were assessed. Multiple procedures were common (an average of 4.5 per patient), and associated with an overall complication rate of 27%. However, after an average follow-up of 5 years, nine of the 12 patients reported improved function and ability to complete normal daily tasks. Most patients were satisfied with their level of pain relief, their grip and pinch strength, and their overall final result. The subjective outcome was less satisfactory in those involved in workers' compensation litigation. All seven attempted scaphoid-thumb metacarpal fusions failed. PMID- 11895357 TI - Synovial chondromatosis of the metacarpophalangeal joint. AB - We describe a case of synovial chondromatosis in a metacarpophalangeal joint with invasion of local structures. The degree of local tissue involvement was not demonstrated on the preoperative MRI scans. This case highlights that synovial chondromatosis can be invasive and that even the best preoperative imaging may not demonstrate this. PMID- 11895356 TI - Spongy hydroxyapatite in hand surgery--a five year follow-up. AB - Twenty-two patients were treated for enchondromas or cysts of the hand by curettage and complete filling of the defect with hydroxyapatite. Postoperatively no complications occurred and marginal osseous integration was radiologically complete after 6 to 8 weeks. Patients were re-examined after a minimum of 5 years after operation and the functional and aesthetic results were excellent in all patients. Radionuclide imaging showed identical bone activity to that in the contralateral healthy hand and there were no signs of inflammation. PMID- 11895359 TI - Re: The sequelae of reflex sympathetic dystrophy. PMID- 11895360 TI - The study of single piece metacarpophalangeal implants. PMID- 11895362 TI - Treatment of neuropathy with autogenous saphenous vein wrapping. PMID- 11895365 TI - Genetic and immunologic considerations in autism. AB - According to recent epidemiological surveys, autistic spectrum disorders have become recognized as common childhood psychopathologies. These life-lasting conditions demonstrate a strong genetic determinant consistent with a polygenic mode of inheritance for which several autism susceptibility regions have been identified. Parallel evidence of immune abnormalities in autistic patients argues for an implication of the immune system in pathogenesis. This review summarizes advances in the molecular genetics of autism, as well as recently emerging concerns addressing the disease incidence and triggering factors. The neurochemical and immunologic findings are analyzed in the context of a neuroimmune hypothesis for autism. Studies of disorders with established neuroimmune nature indicate multiple pathways of the pathogenesis; herein, we discuss evidence of similar phenomena in autism. PMID- 11895366 TI - Presenilin-1 protects against neuronal apoptosis caused by its interacting protein PAG. AB - Mutations in the presenilin-1 (PS-1) gene account for a significant fraction of familial Alzheimer's disease. The biological function of PS-1 is not well understood. We report here that the proliferation-associated gene (PAG) product, a protein of the thioredoxin peroxidase family, interacts with PS-1. Microinjection of a plasmid expressing PAG into superior cervical ganglion (SCG) sympathetic neurons in primary cultures led to apoptosis. Microinjection of plasmids expressing wild-type PS-1 or a PS-1 mutant with a deletion of exon 10 (PS1dE10) by themselves had no effect on the survival of primary SCG neurons. However, co-injection of wild-type PS-1 with PAG prevented neuronal death, whereas co-injection with the mutant PS-1 did not affect PAG-induced apoptosis. Furthermore, overexpression of PAG accelerated SCG neuronal death induced by nerve growth factor deprivation. This sensitizing effect was also blocked by wild type PS-1, but not by PS1dE10. These results establish an assay for studying the function of PS-1 in primary neurons, reveal the neurotoxicity of a thioredoxin peroxidase, demonstrate a neuroprotective activity of the wild-type PS-1, and suggest possible involvement of defective neuroprotection by PS-1 mutants in neurodegeneration. PMID- 11895367 TI - High molecular weight complexes of mutant superoxide dismutase 1: age-dependent and tissue-specific accumulation. AB - Mutations in the cytosolic enzyme, superoxide dismutase 1, have been identified as the cause of motor neuron disease in a subset of cases of familial amyotrophic lateral sclerosis. It has been postulated that the injurious property of mutant enzyme resides in its propensity to aggregate or its propensity to catalyze deleterious, copper-mediated, chemistries. Aggregates of SOD1 have been identified, histologically, in neurons and astroglia of the spinal cords of SOD1 linked FALS patients and in transgenic mice that express these mutant proteins. In the present study, we have employed a technique used in detecting and quantifying aggregates of mutant huntingtin (cellulose acetate filtration) to examine the molecular characteristics of mutant SOD1 in three previously characterized transgenic mouse models of FALS. We show that the brains and spinal cords of these mice accumulate mutant SOD1 complexes that can be trapped by cellulose acetate filtration. The relative abundance of these structures increases dramatically with age. Although expressed to the same level in nonnervous tissues, mutant SOD1 was not found in high molecular weight structures. We conclude that some aspect of the biology of neural tissues (in a setting of declining motor neuron function) predisposes to the accumulation of high molecular weight complexes of mutant SOD1. PMID- 11895368 TI - Neurobehavioral and electroencephalographic abnormalities in Ube3a maternal deficient mice. AB - Angelman syndrome (AS), characterized by motor dysfunction, mental retardation, and seizures, is caused by several genetic etiologies involving chromosome 15q11 q13, including mutations of the UBE3A gene. UBE3A encodes UBE3A/E6-AP, a ubiquitin-protein ligase, and shows brain-specific imprinting, with brain expression predominantly from the maternal allele. Lack of a functional maternal allele of UBE3A causes AS. In order to understand the causal relationship between maternal UBE3A mutations and AS, we have constructed a mouse model with targeted inactivation of Ube3a. The inactive allele contains a lacZ reporter gene for analysis of brain-specific imprinting. Maternal, but not paternal, transmission of the targeted allele leads to beta-galactosidase activity in hippocampal and cerebellar neurons. Maternal inheritance of the Ube3a mutant allele also causes impaired performance in tests of motor function and spatial learning, as well as abnormal hippocampal EEG recordings. As predicted from the dependence of UBE3A mediated ubiquitination of p53 on HPV E6 protein, our maternal-deficient mice show normal brain p53 levels. PMID- 11895369 TI - Marmoset fine B cell and T cell epitope specificities mapped onto a homology model of the extracellular domain of human myelin oligodendrocyte glycoprotein. AB - Aberrant association of autoantibodies with myelin oligodendrocyte glycoprotein (MOG), an integral membrane protein of the central nervous system (CNS) myelin, has been implicated in the pathogenesis of multiple sclerosis (MS). Sensitization of nonhuman primates (Callithrix jacchus marmosets) against the nonglycosylated, recombinant N-terminal domain of rat MOG (residues 1-125) reproduces an MS-like disease in which MOG-specific autoantibodies directly mediate demyelination. To assess the interrelationship between MOG structure and the induction of autoimmune CNS diseases and to enable structure-based rational design of therapeutics, a homology model of human MOG(2-120) was constructed based on consensus residues found in immunoglobulin superfamily variable-type proteins having known structures. Possible sites for posttranslational modifications and dimerization have also been identified and analyzed. The B cell and T cell epitopes have been identified in rat MOG-immunized marmosets, and these sequences are observed to map primarily onto accessible regions in the model, which may explain their ability to generate potent antibody responses. PMID- 11895370 TI - Destabilization of neuronal calcium homeostasis by factors secreted from choroid plexus macrophage cultures in response to feline immunodeficiency virus. AB - The choroid plexus contains a major reservoir of macrophages poised for efficient delivery of virus and neurotoxins to the brain after infection by lentiviruses such as human or feline immunodeficiency virus (FIV). However, their contribution to neurotoxicity is poorly understood. Medium from FIV-infected, choroid plexus macrophages applied to cultured feline cortical neurons induced a small acute calcium rise followed by either a delayed calcium deregulation (41%) or swelling and bursting (23%). NMDA glutamate receptor blockade prevented the acute calcium increase and antagonists to the IP(3) receptor, voltage-gated calcium channels and sodium channels suppressed both the acute and late increases. Analysis of intracellular calcium recovery in toxin-treated neurons after a brief exposure to glutamate, revealed a decrease in the rate and extent of recovery. The apparent diverse pharmacological contributions to intracellular calcium destabilization may be due to the ability of macrophage toxins to interfere with recovery of intracellular calcium homeostasis. PMID- 11895371 TI - Neuropathological and behavioral consequences of adeno-associated viral vector mediated continuous intrastriatal neurotrophin delivery in a focal ischemia model in rats. AB - Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) were continuously delivered to the striatum at biologically active levels via recombinant adeno-associated viral (rAAV) gene transfer 4-5 weeks prior to 30 min of middle cerebral artery occlusion (MCAO). The magnitude of the deficits in a battery of behavioral tests designed to assess striatal function was highly correlated to the extent of ischemic damage determined by unbiased stereological estimations of striatal neuron numbers. The delivery of neurotrophins lead to mild functional improvements in the ischemia-induced motor impairments assessed 3 5 weeks after the insult, in agreement with a small but significant increase of the survival of dorsolateral striatal neurons. Detailed phenotypic analysis demonstrated that the parvalbumin-containing interneurons were spared to a greater extent by the neurotrophin treatment as compared to the projection neurons, which agreed with the specificity for interneuron transduction by the rAAV vector. These data show the advantage of the never previously performed combination of precise quantification of the ischemia-induced neuropathology along with detailed behavioural analysis for assessing neuroprotection after stroke. We observe that intrastriatal delivery of NGF and BDNF using a viral vector system can mitigate, albeit only moderately, neuronal death following stroke, which leads to detectable functional sparing. PMID- 11895372 TI - Ethanol-induced caspase-3 activation in the in vivo developing mouse brain. AB - Recently several methods have been described for triggering extensive apoptotic neurodegeneration in the developing in vivo mammalian brain. These methods include treatment with drugs that block NMDA glutamate receptors, drugs that promote GABA(A) neurotransmission, or treatment with ethanol, which has both NMDA antagonist and GABAmimetic properties. A single intoxication episode induced by any of these agents is sufficient to cause widespread neurodegeneration throughout many brain regions. The cell death process transpires rapidly from early to late stages within several hours. As the neurons die, they become TUNEL positive and show, by both light and electron microscopy, all of the classical morphological characteristics of apoptosis. In the present study, using immunocytochemical methods, we document that ethanol intoxication of 7-day-old infant mice causes a widespread pattern of caspase-3 activation corresponding to the pattern of apoptotic neurodegeneration that is occurring simultaneously. PMID- 11895373 TI - Cisplatin-induced apoptosis of DRG neurons involves bax redistribution and cytochrome c release but not fas receptor signaling. AB - Cisplatin causes apoptosis of DRG neurons in vitro and in vivo that can be prevented by high dose NGF. Design of a neuronal rescue strategy for patients receiving cisplatin will be facilitated by knowledge of the mechanism by which cisplatin causes DRG death. Inhibition of the fas receptor/ligand interaction prevents apoptosis in certain cancer cell lines treated with DNA damaging agents, including cisplatin. We demonstrated that killing curves from mice lacking a functional fas receptor and wild-type controls were not different over a wide range of therapeutically relevant concentrations. However, cisplatin treatment of DRG caused redistribution of cytosolic bax and mitochondrial release of cytochrome c. Bax redistribution was prevented by high dose NGF. This demonstrates for the first time that cisplatin does not signal for death via the fas pathway, but it does initiate the mitochondrial stress pathway in neurons and that NGF blocks death upstream of bax redistribution. PMID- 11895374 TI - Ca(2+) influx through AMPA or kainate receptors alone is sufficient to initiate excitotoxicity in cultured oligodendrocytes. AB - Oligodendrocytes are vulnerable to excitotoxic insults mediated by AMPA receptors and by low and high affinity kainate receptors, a feature that is dependent on Ca(2+) influx. In the current study, we have analyzed the intracellular concentration of calcium [Ca(2+)](i) as well as the entry routes of this cation, upon activation of these receptors. Selective activation of either receptor type resulted in a substantial increase (up to fivefold) of [Ca(2+)](i), an effect which was totally abolished by the non-NMDA receptor antagonist CNQX or by removing Ca(2+) from the culture medium. Blockade of voltage-gated Ca(2+) channels with La(3+) or nifedipine, reduced the amplitude of the Ca(2+) current triggered by AMPA receptor activation by approximately 65%, but not that initiated by low and high affinity kainate receptors. In contrast, KB-R7943, an inhibitor of the plasma membrane Na(+)-Ca(2+) exchanger, solely attenuated the rise in [Ca(2+)](i) by approximately 25% due to activation of low affinity kainate receptors. However, oligodendroglial death by glutamate receptor overactivation was largely unaffected in the presence of La(3+) or KB-R7943. These findings indicate that Ca(2+) influx via AMPA and kainate receptors alone is sufficient to initiate cell death in oligodendrocytes, which does not require the entry of calcium via other routes such as voltage-activated calcium channels or the plasma membrane Na(+)-Ca(2+) exchanger. PMID- 11895375 TI - Systemic increase of oxidative nucleic acid damage in Parkinson's disease and multiple system atrophy. AB - 8-hydroxy-2'-deoxyguanosine (8-OHdG) or 8-hydroxyguanosine (8-OHG), a product of oxidized DNA or RNA, is a good marker of oxidative cellular damage. In this study, we measured the 8-OHdG/8-OHG levels in the serum and cerebrospinal fluid (CSF) of patients with Parkinson's disease (PD) and multiple system atrophy (MSA). Compared to age-matched controls, the mean levels of serum 8-OHdG/8-OHG were significantly higher in PD (P < 0.0001). Although no gender differences were observed in the controls, the mean values of serum 8-OHdG/8-OHG were significantly higher in female PD cases (P < 0.005) than in male patients. 8 OHdG/8-OHG levels in CSF were also increased significantly in patients with PD and MSA, however, their relative values were generally much lower than those in the serum. Together with previous studies showing increased peripheral 8-OHdG levels in Alzheimer's disease and amyotrophic lateral sclerosis, the data presented here suggest that systemic DNA/RNA oxidation is commonly observed in neurodegenerative diseases. Our results also imply that female patients with PD show higher levels of oxidative stress, which may explain the faster progression of this disease in females. PMID- 11895376 TI - Neurofilament-immunoreactive neurons in Alzheimer's disease and dementia with Lewy bodies. AB - The cortical neurons thought to be selectively affected in dementia with Lewy bodies (DLB) are those containing nonphosphorylated 200-kDa neurofilament (NF) protein. As these neurons are largely spared in Alzheimer's disease (AD), DLB and AD may impact on different cortical neuronal populations. The present study quantifies the NF-containing neurons in frontal and temporal cortex of 8 AD, 8 DLB, and 8 control cases. Formalin-fixed paraffin-embedded tissue was immunohistochemically stained with antibodies against nonphosphorylated and phosphorylated NF. Immunoreactive neurons were quantified by areal fraction analysis and corrected for cortical volume. As expected, nonphosphorylated and phosphorylated NF accumulated in the pathological hallmarks of AD and DLB. However, rather than a decrease in NF-containing neurons, a doubling of this population was observed in DLB, compared with AD and controls. This increased number of cortical NF-containing neurons reveal novel widespread cortical changes, beyond those explained by Lewy body formation, that are specific for DLB. PMID- 11895377 TI - Sodium currents in striatal neurons from dystonic dt(sz) hamsters: altered response to lamotrigine. AB - Dystonic mutant dt(sz) hamsters are a model for paroxysmal dystonia. Handling/stress provoke the dystonic attacks. This phenomenon subsedes with maturation, but can be reinvoked when these animals receive sodium channel blockers such as lamotrigine, suggesting a dysfunction of striatal sodium channels. Voltage-gated fast sodium currents (I(Na(+))) were studied in acutely isolated striatal neurons from healthy and dt(sz) hamsters in whole-cell voltage clamp recordings. The action of lamotrigine was tested on (a) current/voltage relationship, (b) kinetics, and (c) steady-state inactivation and activation. Under control conditions, properties of I(Na(+)) were not different between healthy and dt(sz) neurons. With lamotrigine, however, (a) peak currents were significantly less depressed by the drug in neurons from dt(sz) hamsters as compared to healthy cells, and (b) the steady-state inactivation curve shift of I(Na(+)) was less pronounced in dt(sz) neurons. The results suggest that in dt(sz) hamsters, fast sodium currents in striatal neurons are more resistant to blockade. This sodium channel alteration might be causal for a functional imbalance between input and output structures of the basal ganglia under conditions of compromised I(+)(Na). PMID- 11895378 TI - A presenilin 1 mutation associated with familial frontotemporal dementia inhibits gamma-secretase cleavage of APP and notch. AB - A novel presenilin 1 mutation, insR352, associated with a frontal temporal dementia phenotype has been identified (E. A. Rogaeva et al., 2001, Neurology 57, 621-625). This mutation does not increase Abeta42 levels, but instead acts as dominant negative presenilin, decreasing amyloid beta protein (Abeta) production by inhibiting gamma-secretase cleavage of the Abeta precursor. The distinct clinical phenotype associated with this mutation suggests that chronic partial inhibition of gamma-secretase activity may result in neurodegeneration. PMID- 11895379 TI - Structural instability, multiple stable states, and hysteresis in periphyton driven by phosphorus enrichment in the Everglades. AB - Periphyton is a key component of the Everglades ecosystems. It is a major primary producer, providing food and habitat for a variety of organisms, contributing material to the surface soil, and regulating water chemistry. Periphyton is sensitive to the phosphorus (P) supply and P enrichment has caused dramatic changes in the native Everglades periphyton assemblages. Periphyton also affects P availability by removing P from the water column and depositing a refractory portion into sediment. A quantitative understanding of the response of periphyton assemblages to P supply and its effects on P cycling could provide critical supports to decision making in the conservation and restoration of the Everglades. We constructed a model to examine the interaction between periphyton and P dynamics. The model contains two differential equations: P uptake and periphyton growth are assumed to follow the Monod equation and are limited by a modified logistic equation. Equilibrium and stability analyses suggest that P loading is the driving force and determines the system behavior. The position and number of steady states and the stability also depend upon the rate of sloughing, through which periphyton deposits refractory P into sediment. Multiple equilibria may exist, with two stable equilibria separated by an unstable equilibrium. Due to nonlinear interplay of periphyton and P in this model, catastrophe and hysteresis are likely to occur. PMID- 11895380 TI - The metapopulation dynamics of an infectious disease: tuberculosis in possums. AB - An SEI metapopulation model is developed for the spread of an infectious agent by migration. The model portrays two age classes on a number of patches connected by migration routes which are used as host animals mature. A feature of this model is that the basic reproduction ratio may be computed directly, using a scheme that separates topography, demography, and epidemiology. We also provide formulas for individual patch basic reproduction numbers and discuss their connection with the basic reproduction ratio for the system. The model is applied to the problem of spatial spread of bovine tuberculosis in a possum population. The temporal dynamics of infection are investigated for some generic networks of migration links, and the basic reproduction ratio is computed-its value is not greatly different from that for a homogeneous model. Three scenarios are considered for the control of bovine tuberculosis in possums where the spatial aspect is shown to be crucial for the design of disease management operations. PMID- 11895381 TI - Neutral evolution in spatially continuous populations. AB - We introduce a general recursion for the probability of identity in state of two individuals sampled from a population subject to mutation, migration, and random drift in a two-dimensional continuum. The recursion allows for the interactions induced by density-dependent regulation of the population, which are inevitable in a continuous population. We give explicit series expansions for large neighbourhood size and for low mutation rates respectively and investigate the accuracy of the classical Malecot formula for these general models. When neighbourhood size is small, this formula does not give the identity even over large scales. However, for large neighbourhood size, it is an accurate approximation which summarises the local population structure in terms of three quantities: the effective dispersal rate, sigma(e); the effective population density, rho(e); and a local scale, kappa, at which local interactions become significant. The results are illustrated by simulations. PMID- 11895382 TI - Spatial mosaic and interfacial dynamics in a Mullerian mimicry system. AB - Uncovering why spatial mosaics of mimetic morphs are maintained in a Mullerian mimicry system has been a challenging issue in evolutionary biology. In this article, we analyze the reaction diffusion system that describes two-species Mullerian mimicry in one- and two-dimensional habitats. Due to positive frequency dependent selection, a local population first approaches the state where one of the comimicking patterns predominates, which is followed by slow movement of boundaries where different patterns meet. We then analyze the interfacial dynamics of the boundaries to find whether a stable cline is maintained and to obtain the wave speed if the cline is unstable. The results are: (1) In a spatially uniform habitat the morph with greater base fitness spreads both in one and two species system. (2) The strength of cross-species interaction determines whether the mimetic morph clines of model and mimic species coalesce into the same geographical region or pass through each other. The joint wave speed of clines decreases by increasing the number of comimicking species in the mimicry ring. (3) In spatial heterogeneous habitats, stable clines can be maintained due to the balance between the base fitness gradient and the biased gene flow by negative curvature of boundary. This allows the persistence of a spatial mosaic even if one of the morphs is in every place advantageous over the other. A balanced cline is also maintained if there is a gradient in the population density. (4) A new advantageous morph occurring at a local region is doomed to go to extinction in a finite time if the "radius" of initial distribution is below a threshold. Possible applications to the heliconiine butterfly mimicry ring, heterozygous disadvantage systems of chromosomal rearrangement and hybrid zone, the third phase of Wright's Shifting Balance theory, and cytoplasmic incompatibility are discussed. PMID- 11895383 TI - On Richerson and Boyd's model of cultural evolution by sexual selection. AB - Richerson and Boyd proposed a model of sexual selection in which both the male trait and the female preference are culturally transmitted. We generalize their model by introducing sex-dependent oblique transmission rates and a fairly comprehensive female preference rule. The model differs markedly from the standard genetic models in that the male trait and the female preference are uncorrelated. Hence, there can be no "sexy son" effect to offset the assumed fertility cost to choosy females. Nevertheless, the cultural processes can support a stable polymorphic equilibrium at which the choosy females are present. Also of interest are the cyclical dynamics observed in the neighborhood of the internal equilibrium. PMID- 11895384 TI - Twin studies in behavioral research: a skeptical view. AB - We review in detail two major ongoing research projects that employ samples of twins reared apart (and in one case, twins reared together). The studies attempt, via model fitting, to estimate proportions of genetic and environmental variance for many human traits. We discuss problems concerning the representativeness of samples, the accuracy and reliability of the data, the extent of contact of nominally separated twins, the measurement of selective placement effects, and the particular model-fitting procedures. The two studies agree in their conclusions, but we do not find the conclusions to be convincing. We suggest that no scientific purpose is served by the flood of heritability estimates generated by these studies. PMID- 11895385 TI - The preorganization step in organic reaction mechanisms. Charge-transfer complexes as precursors to electrophilic aromatic substitutions. AB - Metastable (pre-reactive) intermediates, as commonplace transients in simple bimolecular reactions, are usually unobserved (and ignored)-though they provide vital mechanistic insight. Thus, the preequilibrium (charge-transfer) complexes of various aromatic donors with rather typical electron acceptors such as Br(2), NO(+), and NO(2)(+) are examined quantitatively (via their molecular and electronic structures) to reveal surprisingly unorthodox aspects of what is conventionally referred to in organic chemistry textbooks as electrophilic aromatic bromination, nitrosation, and nitration, respectively. PMID- 11895387 TI - Directing abilities of alcohol-derived functional groups in the hydroformylation of olefins. AB - The hydroformylation of allylic and homoallylic alcohols and their derivatives using cationic and neutral rhodium complexes has been examined. The highest diastereoselectivity (87:13) was observed in the reaction of 1-methoxymethoxy-2 methylenecyclohexane. Higher yields and similar selectivities were obtained in the reaction of the TBDMS-protected alcohol. The major diastereomer results from hydroformylation syn to the functional group, which would suggest a directing effect. However, hydroformylation of 3-methylene-1-cyclohexanol derivatives occurs on the face opposite to the directing group in the major isomer. These data, in addition to the results of hydroformylation of 1-methyl-2 methylenecyclohexane, suggest that inherent conformational preferences are of significant importance in determining the product distribution and that the directing power of simple alcohols and their derivatives is moderate at best under the conditions examined in this study. PMID- 11895386 TI - Free-radical-mediated conjugate additions. Enantioselective synthesis of butyrolactone natural products: (-)-enterolactone, (-)-arctigenin, (-) isoarctigenin, (-)-nephrosteranic acid, and (-)-roccellaric acid. AB - Lewis acid-mediated conjugate addition of alkyl radicals to a differentially protected fumarate 10 produced the monoalkylated succinates with high chemical efficiency and excellent stereoselectivity. A subsequent alkylation or an aldol reaction furnished the disubstituted succinates with syn configuration. The chiral auxiliary, 4-diphenylmethyl-2-oxazolidinone, controlled the stereoselectivity in both steps. Manipulation of the disubstituted succinates obtained by alkylation furnished the natural products (-)-enterolactone, (-) arctigenin, and (-)-isoarctigenin. The overall yields for the target natural products were 20-26% over six steps. Selective functionalization of the disubstituted succinates obtained by aldol condensation gave the paraconic acid natural products (-)-nephrosteranic acid (8) and (-)-roccellaric acid (9). The overall yield of the natural products 8 and 9 over four steps was 53% and 42%, respectively. PMID- 11895388 TI - Mechanistic study of the Mitsunobu reaction. AB - The Mitsunobu reaction occurs typically with inversion of configuration in secondary alcohol derivatives. In this paper, a mechanistic explanation for lactonizations of hindered alcohols under Mitsunobu conditions with retention is proposed. This involves the intermediacy of an acyloxyphosphonium salt followed by acyl transfer to the alcohol. PMID- 11895389 TI - An approach to the stereoselective synthesis of syn- and anti-1,3-diol derivatives. Retention of configuration in the Mitsunobu reaction. AB - The Mitsunobu reaction typically proceeds with inversion of configuration at the hydroxyl center. However, with a series of hindered alcohols, the intramolecular version of the Mitsunobu reaction afforded exclusively the product of retention of configuration. A mechanistic rationale for this observation is discussed, wherein this atypical stereochemical outcome is attributed to steric congestion at the reaction center. PMID- 11895391 TI - Concise syntheses of nonracemic gamma-fluoroalkylated allylic alcohols and amines via an enantiospecific palladium-catalyzed allylic substitution reaction. AB - Alpha-fluoroalkylated allyl mesylates reacted with various carboxylates and amines in the presence of tetrakis(triphenylphosphine)palladium(0) catalyst to give the corresponding gamma-fluoroalkylated (E)-allylic alcohol derivatives and amines, respectively, in excellent yields. In almost all cases, no other regio- and stereoisomers were produced. Application of this palladium-catalyzed allylic substitution reaction to various nonracemic mesylates afforded chiral gamma fluoroalkylated allylic alcohol derivatives and amines without any loss of enantiomeric excess through the reaction. PMID- 11895392 TI - Diphenyl disulfide and sodium in NMP as an efficient protocol for in situ generation of thiophenolate anion: selective deprotection of aryl alkyl ethers and alkyl/aryl esters under nonhydrolytic conditions. AB - Aryl methyl ethers, methyl esters, aryl esters, and aryl sulfonates are chemoselectively deprotected under nonhydrolytic conditions by treatment with Ph(2)S(2) (0.6 equiv) and Na (1.6 equiv) in NMP under reflux or at 90 degrees C. Quantitative utilization of the 'PhS' moiety as the effective nucleophilic species represents conservation of atom economy. Other solvents such as HMPA, DMPU, DMEU, and DMF afforded comparable results. Chloro, nitro, aldehyde, alpha,alpha-diketone, and alpha,beta-unsaturated ketone functionalities remain unaffected. The deprotection was found to take place in the order aryl ester > alkyl ester > aryl alkyl ether. Substrates bearing strong electron-withdrawing groups react at a faster rate than those not having such substitution. The differences in rate of reaction has been exploited for selective deprotection for intramolecular competition. An aryl acetate/benzoate is deprotected selectively in preference to a methyl ester or aryl methyl ether. Selective deprotection of a methyl ester is observed in the presence of an aryl alkyl ether. PMID- 11895390 TI - Kulokekahilide-1, a cytotoxic depsipeptide from the cephalaspidean mollusk Philinopsis speciosa. AB - The cytotoxic depsipeptide kulokekahilide-1, which contains two unusual amino acids, 4-phenylvaline and 3-amino-2-methylhexanoic acid, was isolated from the cephalaspidean mollusk Philinopsis speciosa. Structure elucidation of kulokekahilide-1 was carried out by spectroscopic analysis and chemical degradation. The absolute stereochemistry was determined by Marfey analysis for amino acids and chiral HPLC analysis for hydroxy acids. All four stereoisomers of 4-phenylvaline and 3-amino-2-methylhexanoic acid, which were necessary for Marfey analysis, were synthesized by use of the Heck reaction and Evans's method, respectively. Kulokekahilide-1 showed cytotoxicity against P388 murine leukemia cells with an IC(50) value of 2.1 microg/mL. PMID- 11895394 TI - [3 + 2] Annulation of beta-heteroatom-substituted alpha,beta-unsaturated acylsilanes with methyl ketone enolates: scope and investigation of the reaction course. AB - A new route to (Z)-beta-silylacryloylsilanes 10 and the improved conditions for the [3 + 2] annulation using 10 and alkyl methyl ketone enolates are reported. Also, details of investigations defining a reaction course of the [3 + 2] annulation using beta-phenylthio- and beta-trimethylsilyl-acryloylsilanes 1 (X = SPh, SiMe(3)) and alkyl methyl ketone enolates are described. PMID- 11895393 TI - Combining lipase-catalyzed enantiomer-selective acylation with fluorous phase labeling: a new method for the resolution of racemic alcohols. AB - Lipase-catalyzed acylation of racemic alcohols with a highly fluorinated acyl donor allows their kinetic resolution accompanied by the simultaneous enantiomer selective fluorous phase labeling. Both the tagged and the untagged enantiomer can be separated without chromatography by a very efficient partition between a fluorous and an organic phase. The method has been successfully applied to the resolution of typically racemic secondary alcohols of low molecular weight. The fluorous label can be recovered quantitatively. PMID- 11895395 TI - Elongation and contraction of molecular springs. Synthesis, structures, and properties of bridged [7]thiaheterohelicenes. AB - A series of bridged [7]thiaheterohelicenes 3a-c and 4 with a variety of helical pitches have been prepared from racemic and optical pure 2,13 bis(hydroxymethyl)dithieno[3,2-e:3',2'-e']benzo[1,2-b:4,3-b']bis[1]benzothiophene (1) in order to investigate the helical structures in solution. Recrystallizations of (PM)-3a, (PM)-3b, (PM)-3c, and (P)-4 from hexane dichloromethane gave crystals suitable for X-ray crystallography, while recrystallization of (PM)-4 with benzene gave an inclusion complex with a stoichiometry of (PM-4)(4).(C(6)H(6)). X-ray analyses of (PM)-3a-c, (PM 4)(4).(C(6)H(6)), and (P)-4 indicate that the dihedral angles between terminal thiophene rings of the helical framework significantly vary from 22 degrees for 4 to 59 degrees for 3c. This represents as increase of 37 degrees or 168%. Although the (13)C NMR and UV absorption spectra of bridged helicenes 3a-c and unbridged helicene 5 are essentially the same, the molar rotation of 5 is very large compared with those of 3a-c and 4. A red shift (15 nm) in the circular dichroism (CD) spectrum is observed for 4, suggesting that this compound is more planar than 3a-c in solution. In the series of [7]thiaheterohelicenes studied, the minimum helical pitch is 2.70 A for 4. PMID- 11895396 TI - Enantiomerically pure alpha-amino acid synthesis via hydroboration-suzuki cross coupling. AB - The Garner aldehyde-derived methylene alkene 5 and the corresponding benzyloxycarbonyl compound 25 undergo hydroboration with 9-BBN-H followed by palladium-catalyzed Suzuki coupling reactions with aryl and vinyl halides. After one-pot hydrolysis-oxidation, a range of known and novel nonproteinogenic amino acids were isolated as their N-protected derivatives. These novel organoborane homoalanine anion equivalents are generated and transformed under mild conditions and with wide functional group tolerance: electron-rich and -poor aromatic iodides and bromides (and a vinyl bromide) all undergo efficient Suzuki coupling. The extension of this methodology to prepare meso-DAP, R,R-DAP, and R,R-DAS is also described. PMID- 11895398 TI - Five- and six-membered ring opening of pyroglutamic diketopiperazine. AB - A variety of ring-opening reactions of pyroglutamic diketopiperazine at both the five-membered and six-membered rings is described. Mild, basic conditions facilitate nucleophilic attack by amines at the diketopiperazine carbonyls giving pyroglutamides in excellent yield. Reaction with nucleophiles under acidic conditions give bis-glutamate derivatives of 2,5-diketopiperazine (DKP). These reactions provide simple, two-step sequences to pyroglutamides and symmetrical diketopiperazines from commercial pyroglutamic acid with control of product dictated by reaction conditions, catalyst, and nucleophile. PMID- 11895397 TI - Stereodefined synthesis of O3'-labeled uracil nucleosides. 3'-[(17)O]-2'-Azido-2' deoxyuridine 5'-diphosphate as a probe for the mechanism of inactivation of ribonucleotide reductases. AB - Thermolysis of a 2'-[(16)O]-O-benzoyl-[(17)O]-5'-O-(tert-butyldimethylsilyl) O(2),3'-cyclouridine derivative gave the more stable 3'-[(17)O]-O-benzoyl-[(16)O] 5'-O-(tert-butyldimethylsilyl)-O(2),2'-cyclouridine isomer, which was converted into 3'-[(17)O]-2'-azido-2'-deoxyuridine by deprotection and nucleophilic ring opening at C2' with lithium azide. The 5'-diphosphate was prepared by nucleophilic displacement of the 5'-O-tosyl group with tris(tetrabutylammonium) hydrogen pyrophosphate. Model reactions gave (16)O and (18)O isotopomers, and base-promoted hydrolysis of an O(2),2'-cyclonucleoside gave stereodefined access to 3'-[(18)O]-1-(beta-D-arabinofuranosyl)uracil. Inactivation of ribonucleoside diphosphate reductase with 2'-azido-2'-deoxynucleotides results in appearance of EPR signals for a nitrogen-centered radical derived from azide, and 3'-[(17)O]-2' azido-2'-deoxyuridine 5'-diphosphate provides an isotopomer to perturb EPR spectra in a predictable manner. PMID- 11895399 TI - Theoretical and experimental studies of biotin analogues that bind almost as tightly to streptavidin as biotin. AB - We have used a newly developed qualitative computational approach, PROFEC (Pictorial Representation of Free Energy Changes), to visualize the areas of the ligand biotin where modifications of its structure might lead to tighter binding to the protein streptavidin. The PROFEC analysis, which includes protein flexibility and ligand solvation/desolvation, led to the suggestion that the pro 9R hydrogen atom of biotin, which is in alpha-position to the CO(2)(-) group, might be changed to a larger group and lead to better binding with streptavidin and avidin. Free energy calculations supported this suggestion and predicted that the methyl analogue should bind approximately 3 kcal/mol more tightly to streptavidin, with this difference coming exclusively from the relative desolvation free energy of the ligand. The PROFEC analysis further suggested little or no improvement for changing the pro-9S hydrogen atom to a methyl group, and great reduction in changing the ureido N-H groups to N-CH(3). Stimulated by these results, we synthesized 9R-methylbiotin and 9S-methylbiotin, and their binding free energies and enthalpies were measured for interaction with streptavidin and avidin, respectively. In contrast to the calculated results, experiments found both 9-methylbiotin isomers to bind more weakly to streptavidin than biotin. The calculated preference for the binding of the 9R- over the 9S stereoisomer was observed. In addition, 9-methylbiotin is considerably less soluble in water than biotin, as predicted by the calculation, and the 9R isomer is, to our knowledge, thus far the tightest binding analogue of biotin to streptavidin. Subsequently, X-ray structures of the complexes between streptavidin and both 9R- and 9S-methylbiotin were determined, and the structures were consistent with those used in the free energy calculations. Thus, the reason for the discrepancy between the calculated and experimental binding free energy does not lie in unusual binding modes for the 9-methylbiotins. PMID- 11895400 TI - Electrochemical homocoupling of 2-bromomethylpyridines catalyzed by nickel complexes. AB - 2,2'-Bipyridine (bpy) and a series of dimethyl-2,2'-bipyridines were synthesized from 2-bromopyridine and 2-bromomethylpyridines, respectively, using an electrochemical process catalyzed by nickel complexes. The method is simple and efficient, with isolated yields between 58 and 98% according to the structure. We first studied the influence of the presence and the position of the methyl group on the yield, using N,N-dimethylformamide (DMF) or acetonitrile (AN) as the solvent, NiBr(2)bpy as the catalyst, and Zn as the sacrificial anode, in an undivided cell and at ambient temperature. On the basis of a better understanding of the reaction mechanism based on electroanalytical studies, we could improve the dimerization both by substituting the catalyst ligand (bpy) by the reagent itself, i.e., 2-bromomethylpyridine or 2-bromopyridine, and by using Fe instead of Zn as the sacrificial anode. PMID- 11895401 TI - A novel photochemical reaction of 2-alkoxynicotinates to cage-type photodimers. AB - Irradiation of a benzene solution of 2-alkoxynicotinic acid alkyl esters gave cage-type photodimers in good yields, the structure of which was established by X ray single-crystal analysis. The maximum quantum yield was 8.0 x 10(-)(2) when a 5.0 M (almost neat) solution was used. Photolysis of phenyl 2-methoxynicotinate promoted photo-Fries rearrangement to give 1,3- and 1,5-rearranged products. Excimer emission of methyl 2-methoxynicotinate was observed at 77 K. PMID- 11895402 TI - A new type of ionophore family utilizing the cation-olefinic pi interaction: theoretical study of [n]beltenes. AB - The possible utilization of [n]beltenes as a new family of ionophores, which exhibit a cation-olefinic pi type of interaction in contrast to the cation aromatic pi type of interaction exhibited by [n]collarenes, has been investigated using both ab initio calculations and molecular dynamic simulations. Like [n]collarenes, n ethene groups are linked by -CH(2)- linkages in the [n]beltenes. Our calculations indicate that these [n]beltenes exhibit strong binding affinities and high selectivity for alkali metal cations ([5]beltene to Li(+), [6]beltene to Na(+), [7]beltene to Na(+) and K(+), [8]beltene to K(+) and Rb(+), and [9]beltene to Cs(+) and Rb(+)). Compared to [n]collarenes, [n]beltenes are expected to have a finer ion selectivity because their cavity sizes can be varied with integral number n, while that of the former can be varied with an even number n. Suitable substituents could be employed to enhance both the binding and specificity of various sizes of [n]beltenes to different cations, as well as to increase the solubility. PMID- 11895403 TI - The hydrogen bonding and hydration of 2'-OH in adenosine and adenosine 3'-ethyl phosphate. AB - The 2'-OH group has major structural implications in the recognition, processing, and catalytic properties of RNA. We report here intra- and intermolecular H bonding of 2'-OH in adenosine 3'-ethyl phosphate (1), 3'-deoxyadenosine (2), and adenosine (3) by both temperature- and concentration-dependent NMR studies, as well as by detailed endo ((3)J(H,H)) and exocyclic ((3)J(H,OH)) coupling constant analyses. We have also examined the nature of hydration and exchange processes of 2'-OH with water by a combination of NOESY and ROESY experiments in DMSO-d(6) containing 2 mol % HOD. The NMR-constrained molecular modeling (by molecular mechanics as well as by ab initio methods both in the gas and solution phase) has been used to characterize the energy minima among the four alternative dihedrals possible from the solution of the Karplus equation for (3)J(H2',OH) and (3)J(H3',OH) to delineate the preferred orientation of 2'-O-H proton in 1 and 2 as well as for 2'/3'-O-H protons in 3. The NMR line shape analysis of 2'-OH gave the DeltaG(H-bond)(298K) of 7.5 kJ mol(-1) for 1 and 8.4 kJ mol(-1) for 3; similar analyses of the methylene protons of 3'-ethyl phosphate moiety in 1 also gave comparable DeltaG(H-bond)(298K) of 7.3 kJ mol(-1). The donor nature of the 2'-OH in the intramolecular H-bonding in 3 is evident from its relatively reduced flexibility [-TDeltaS++](2'-OH) = -17.9(+/-0.5) kJ mol(-1)] because of the loss of conformational freedom owing to the intramolecular 2'O-H...O3' H-bonding, compared to the acceptor 3'-OH in 3 [-TDeltaS++](3'-OH) = -19.8 (+/- 0.6) kJ mol( 1)] at 298 K. The presence of intramolecular 2'-OH...O3' H-bonding in 3 is also corroborated by the existence of weak long-range (4)J(H2',OH3') in 3 (i.e., W conformation of H2'-C2'-C3'-O3'-H) as well as by (3)J(H,OH) dependent orientation of the 2'- and 3'-OH groups. The ROESY spectra for 1 and 3 at 308 K, in DMSO d(6), show a clear positive ROE contact of both 2'- and 3'-OH with water. The presence of a hydrophilic 3'-phosphate group in 1 causes a much higher water activity in the vicinity of its 2'-OH, which in turn causes the 2'-OH to exchange faster, culminating in a shorter exchange lifetime (tau) for 2'-OH proton with HOD in 1 (tau2'-OH: 489 ms) compared to that in 3 (tau2'-OH: 6897 ms). The activation energy (E(a)) of the exchange with the bound-water for 2'- and 3'-OH in 3 (48.3 and 45.0 kJ mol(-1), respectively) is higher compared to that of 2'-OH in 1 (31.9 kJ mol(-1)), thereby showing that the kinetic availability of hydrated 2'-OH in 1 for any inter- and intramolecular interactions, in general, is owing to the vicinal 3'-phosphate residue. It also suggests that 2'-OH in native RNA can mediate other inter- or intramolecular interactions only in competition with the bound-water, depending upon the specific chemical nature and spatial orientation of other functions with potential for hydrogen bonding in the neighborhood. This availability of the bound water around 2'-OH in RNA would, however, be dictated by whether the vicinal phosphate is exposed to the bulk water or not. This implies that relatively poor hydration around a specific 2'-OH across a polyribonucleotide chain, owing to some hydrophobic microenvironmental pocket around that hydroxyl, may make it more accessible to interact with other donor or acceptor functions for H-bonding interactions, which might then cause the RNA to fold in a specific manner generating a new motif leading to specific recognition and function. Alternatively, a differential hydration of a specific 2'-OH may modulate its nucleophilicity to undergo stereospecific transesterification reaction as encountered in ubiquitous splicing of pre-mRNA to processed RNA or RNA catalysis, in general. PMID- 11895404 TI - Protease-mediated fragmentation of p-amidobenzyl ethers: a new strategy for the activation of anticancer prodrugs. AB - A new anticancer prodrug activation strategy based on the 1,6-elimination reaction of p-aminobenzyl ethers is described. Model studies were undertaken with the N-protected peptide benzyloxycarbonyl-valine-citrulline (Z-val-cit), which was attached to the amino groups of p-aminobenzyl ether derivatives of 1-naphthol and N-acetylnorephedrine. The amide bond that formed was designed for hydrolysis by cathepsin B, a protease associated with rapidly growing and metastatic carcinomas. Upon treatment with the enzyme, the Z-val-cit-p-amidobenzyl ether of 1-naphthol (2) underwent peptide bond hydrolysis with the rapid release of 1 naphthol. The aliphatic Z-val-cit-p-amidobenzyl ether of N-acetylnorephedrine (5) also underwent amide bond hydrolysis, but without the ensuing elimination of N acetylnorephedrine. On the basis of these results, the phenolic anticancer drugs etoposide (6) and combretastatin A-4 (7) were attached to the Z-val-cit-p amidobenzyl alcohol through ether linkages, forming the peptide-drug derivatives 8 and 9, respectively. Both compounds were stable in aqueous buffers and serum and underwent ether fragmentation upon treatment with cathepsin B, resulting in the release of the parent drugs in chemically unmodified forms. The released drugs were 13-50 times more potent than were the prodrug precursors on a panel of cancer cell lines. In contrast, the corresponding carbonate derivative of combretastatin A-4 (13) was unstable in aqueous environments and was as cytotoxic as combretastatin A-4. This result extends the use of the self-immolative p aminobenzyl group for the fragmentation of aromatic ethers and provides a new strategy for anticancer prodrug development. PMID- 11895405 TI - Acid-base equilibria in nonpolar media. 2.(1) Self-consistent basicity scale in THF solution ranging from 2-methoxypyridine to EtP(1)(pyrr) phosphazene. AB - Relative ion-pair basicities Delta(pK)(ip) of 25 substituted aryl and alkyl iminophosphoranes (phosphazenes) and 20 other N-bases (various pyridines, amines, amidines) have been measured in THF medium using the UV-Vis and/or (13)C NMR methods. The Delta(pK)(ip) values were corrected for ion pairing using the Fuoss equation to obtain relative ionic basicities Delta(pK)(alpha). Based on the measurements, a basicity scale ranging from 2-methoxypyridine to EtP(1)(pyrr) and having a total span over 18 pK units has been created. The scale has been anchored to the pK(alpha) value of triethylamine (pK(alpha) = 12.5). The results are compared to pK(a) values in various other solvents and in the gas phase. The pK(alpha) values give better correlations than the pK(ip) values, thus indirectly validating the procedure of correction for ion pairing. The predictability of the basicity together with suitable spectral properties in the UV range make the phenylphosphazenes convenient neutral indicators in the high basicity range where the choice of neutral indicators is very limited. PMID- 11895406 TI - Ab initio and experimental studies on the hetero-Diels-Alder and cheletropic additions of sulfur dioxide to (E)-1-methoxybutadiene: a mechanism involving three molecules of SO(2). AB - Kinetics on the cheletropic addition of sulfur dioxide to (E)-1-methoxybutadiene (1) to give the corresponding sulfolene 2 (2-methoxy-2,5-dihydrothiophene-1,1 dioxide) gave the rate law d[2]/dt = k[1][SO(2)](x)() with x = 2.6 +/- 0.2 at 198 K. Under these conditions, no sultine 3 [(2RS,6RS)-6-methoxy-3,6-dihydro-1,2 oxathiin-2-oxide] resulting from a hetero-Diels-Alder addition was observed, and the cheletropic elimination 2 --> 1 + SO(2) did not occur. Ab initio and DFT quantum calculations confirmed that the cheletropic addition 1 + SO(2) --> 2 follows two parallel mechanisms, one involving two molecules of SO(2) and the transition structure with DeltaG(++) = 18.2 +/- 0.2 kcal/mol at 198 K (exptl); 22.5-22.7 kcal/mol [B3LYP/6-31G(d,p)], the other one involving three molecules of SO(2) with DeltaG(++) = 18.9 +/- 0.1 kcal/mol at 198 K (exptl); 19.7 kcal/mol [B3LYP/6-31G(d,p)]. The mechanism involving only one molecule of SO(2) in the transition structure requires a higher activation energy, DeltaG(++) = 25.2 kcal/mol [B3LYP/6-31G(d,p)]. Comparison of the geometries and energetics of the structures involved into the 1 + SO(2) --> 2, 3 and 1 + 2SO(2) --> 2, 3 + SO(2) reactions obtained by ab initio and DFT methods suggest that the latter calculation techniques can be used to study the cycloadditions of sulfur dioxide. The calculations predict that the hetero-Diels-Alder addition 1 + SO(2) --> 3 also prefers a mechanism in which three molecules of SO(2) are involved in the cycloaddition transition structure. At 198 K and in SO(2) solutions, the entropy cost (TDeltaS(++)) is overcompensated by the specific solvation by SO(2) in the transition structures of both the cheletropic and hetero-Diels-Alder reactions of (E)-1-methoxybutadiene with SO(2). PMID- 11895407 TI - From a biogenetic scenario to a synthesis of the ABC ring of manzamine A. AB - On the basis of a biogenetic proposal for explaining the biogenesis of manzamine A, the cycloaddition of dihydropyridinium salt 26 with diene derivative 5 leads to adducts 27. These adducts, as well as their related and previously described analogues 9, are now shown to be precursors of diene derivatives such as 10, 13, and 28. Treatment of diene 32 with sodium azide resulted in a one-step formation of the tricyclic imino derivative 34. This key intermediate was further transformed into tricyclic derivative 40, which possesses the essential features of the ABC ring of manzamine A. PMID- 11895408 TI - 2-Trimethylsilylethyl sulfides in the von Braun cyanogen bromide reaction: selective preparation of thiocyanates and application to nucleoside chemistry. AB - Mixed 2-(trimethylsilyl)ethyl sulfides were synthesized and used in the von Braun cyanogen bromide reaction for preparing selectively thiocyanates in high yield. We show here that this cleavage reaction is highly selective in methanol in comparison with the reaction of the corresponding non-silyl sulfide analogues. This reaction was applied to the synthesis of nucleosidic thiocyanates such as the new nucleosides 14 and 18 in the search for mechanism-based inhibitors of ribonucleoside diphosphate reductase and bioactive molecules. The selective cleavage is possible for sulfides bearing hydroxyl functions and aromatic rings. The reactions of cyanogen bromide as cyanating and brominating agent were observed for the first time under the same conditions with the naphthoxyhexyl 2 trimethylsilylethyl sulfide 7, which, treated with cyanogen bromide in dichloromethane, led selectively to the p-bromonaphthoxyhexyl thiocyanate 10 in 89% yield. Another reaction induced by cyanogen bromide was observed in dichloromethane with the 2-(trimethylsilylethyl)thio nucleoside 13, which gives the corresponding symmetrical disulfide 21 in good yield. PMID- 11895409 TI - Synthesis of 2,3-disubstituted benzo[b]thiophenes via palladium-catalyzed coupling and electrophilic cyclization of terminal acetylenes. AB - 2,3-Disubstituted benzo[b]thiophenes have been prepared in excellent yields via coupling of terminal acetylenes with commercially available o-iodothioanisole in the presence of a palladium catalyst and subsequent electrophilic cyclization of the resulting o-(1-alkynyl)thioanisole derivatives. I(2), Br(2), NBS, p O(2)NC(6)H(4)SCl, and PhSeCl have been utilized as electrophiles. Aryl-, vinyl-, and alkyl-substituted terminal acetylenes undergo this coupling and cyclization to produce excellent yields of benzo[b]thiophenes. (Trimethylsilyl)acetylene also undergoes this coupling/cyclization process with I(2), NBS, and the sulfur and selenium electrophiles to afford the corresponding 2 (trimethylsilyl)benzo[b]thiophenes. However, cyclization of the silyl-containing thioanisole using Br(2) affords 2,3-dibromobenzo[b]thiophene. PMID- 11895410 TI - Synthesis and structural characterization of cis- and trans-fused 4a,5,6,7,8,8a hexahydro-2H,4H-1,3-benzodithiines and their 2-methyl and 2,2-dimethyl derivatives. AB - Both cis- and trans-fused 4a,5,6,7,8,8a-hexahydro-2H,4H-1,3-benzodithiine together with their 2-methyl and 2,2-dimethyl derivatives were prepared as racemates from the appropriate dithiols obtained via multistep syntheses. The products were characterized by (1)H and (13)C NMR, mass spectrometry, and for two of the cis-fused compounds by X-ray diffraction. (1)H,(1)H vicinal coupling constants indicated that all compounds attain chair-chair conformations as their predominant conformations. All three trans-fused isomers exist in totally biased chair-chair conformations and are essentially conformationally locked, whereas the cis-fused compounds are conformationally mobile and can potentially attain either the S-in or the S-out conformation. The interconversion of the conformers is fast on the NMR time-scale at ambient temperatures, but at 213 K 4ar,5,6,7,8,8ac-hexahydro-1,3-benzodithiine freezes out into a 83:17 mixture of the S-in and S-out forms, respectively. Both 2c-methyl-4ar,5,6,7,8,8ac-hexahydro 1,3-dithiine and the dimethyl derivative adopt almost exclusively the S-in conformer at ambient temperature whereas 2t-methyl-4ar,5,6,7,8,8ac-hexahydro-1,3 dithiine is a 5:1 mixture of the S-out and S-in conformers. PMID- 11895411 TI - Polyfluoroether derivatives via nucleophilic fluorination of glyoxal hydrates with deoxofluor. AB - Various glyoxal hydrates have been reacted with Deoxofluor [(CH(3)OCH(2)CH(2))(2)NSF(3)]. In concentrated solutions of dichloromethane, Deoxofluor (1) efficiently fluorinates a variety of glyoxal hydrates, RCOCHO.H(2)O (R = 4-methoxyphenyl, 3,4-methylenedioxyphenyl, 4-methylphenyl, 4 fluorophenyl, phenyl, 2-thienyl, methyl) (6a-g) to form polyfluoroethers 7a-g and 8a-g as meso and racemic mixtures (approximately 1:1) in good yields. The meso and racemic compounds were separated by flash chromatography and characterized. When the reactant comprised two different glyoxal hydrates, mixed polyfluoroethers (9h-j) were observed as the major products. The yields of the mixed polyfluoroethers depend on the ratio of the two different glyoxal hydrates used. Reactions of some other hydrates, such as hydrindantin dihydrate (10) and 1,1,1,5,5,5-hexafluoro-2,2,4,4-pentanetetrol (11), were also studied with Deoxofluor to give a cyclic polyfluoroether (12) and beta-ketoamine (13), respectively. When the reactions of 6a-d were carried out under very dilute conditions, difluoro aldehydes (14a-d) or tetrafluoroalkanes (15a-d) were formed rather than polyfluoroethers. Reactions of concentrated solutions of nonhydrated glyoxals (16k-m) in methylene chloride with Deoxofluor produced the tetrafluoroalkanes (18k-m) in good yields with only trace amounts of difluoroaldehydes (17k-m) being found. The structures of 7a (meso), 8b (racemic), and 12 have been confirmed by single-crystal X-ray analysis. PMID- 11895412 TI - Metal-mediated carbonyl-1,3-butadien-2-ylation by 1,4-bis(methanesulfonyl)-2 butyne or 1,4-dibromo-2-butyne in aqueous media: asymmetric synthesis of 3 substituted 3-hydroxy-beta-lactams. AB - Metal-mediated 1,3-butadien-2-ylation reactions between 1,4-dibromo-2-butyne or 1,4-bis(methanesulfonyl)-2-butyne and optically pure azetidine-2,3-diones were investigated in aqueous media, offering a convenient asymmetric entry to the potentially bioactive 3-substituted 3-hydroxy-beta-lactam moiety. The diastereoselectivity of the addition reaction was controlled by the bulky chiral auxiliary at C4. However, while the regioselectivity of the process was full, the chemical yield of the addition was a function of the nature of both the metal reagent and the system solvent as well. In addition, 2-azetidinone-tethered 1,3 butadienes can easily be transformed into other functionalities via Diels-Alder reaction. PMID- 11895413 TI - Asymmetric synthesis of axially chiral benzamides and anilides by enantiotopic lithiation of prochiral arene chromium complexes. AB - Axially chiral benzamides and anilides were prepared by enantiotopic lithiation at the distinguished benzylic methyl of prochiral tricarbonylchromium complexes of N,N-diethyl 2,6-dimethylbenzamide (1) and N-methyl-N-acyl 2,6-dimethylaniline (14 and 21) with a chiral lithium amide base followed by electrophilic substitution in good yields with high optical purity. The resulting axially chiral chromium-complexed benzamides and anilides were oxidized under air to give chromium-free axially chiral benzamides and anilides in an enantiomerically active form without axial bond rotation at room temperature. PMID- 11895414 TI - Highly efficient B(C(6)F(5))(3)-catalyzed hydrosilylation of olefins. AB - A convenient and highly efficient method for the Lewis acid-catalyzed trans selective hydrosilylation of alkenes has been developed. The mechanism of this novel protocol operates via direct addition of silylium type species across C=C bond followed by trapping of the resultant carbenium ion with boron-bound hydride. A number of diversely substituted silanes possessing both aryl and alkyl groups at silicon atom were efficiently prepared using this hydrosilylation methodology. The possibility to employ aryl-containing hydrosilanes in this reaction opens broad capabilities for the synthesis of alcohols via a trans selective hydrosilylation/Tamao-Fleming oxidation sequence, complementary to the existing cis-selective hydroboration/oxidation protocol. PMID- 11895415 TI - Synthesis of aza polycyclic compounds derived from pyrrolidine, indolizidine, and indole via intramolecular Diels-Alder cycloadditions of neutral 2-azadienes. AB - A method for the preparation of novel oxaza and diaza polycyclic 9-oxa-4 azaphenanthrene, 5H-pyrido[2,3-a]pyrrolizine, 5H,6H-pyrido[3,2-g]indolizine, and 5H,6H-indeno[2,1-a]indole is described, based on tandem reactions: aza-Wittig reaction of N-vinylic phosphazenes with functionalized aldehydes and an intramolecular aza-Diels-Alder reaction. PMID- 11895416 TI - Remarkable diastereoselectivity in the addition of allylic and unsaturated diorganozinc reagents to beta-(N,N-dialkylamino)-aldehydes. AB - 1,3-Anti amino alcohols 5(a)-18(a) are obtained with high diastereoselectivity by use of diorganozinc reagents in additions to amino aldehydes 2a and 2b. The corresponding Grignard reagents exhibit low to modest diastereoselectivity. The highly diastereoselective zinc-based method makes available a wide range of 4,4 disubstituted cyclohexenone derivatives containing contiguous stereocenters. PMID- 11895417 TI - Gas-phase identity nucleophilic substitution reactions of cyclopropenyl halides. AB - The gas-phase identity nucleophilic substitution reactions of halide anions (X = F, Cl, and Br) with cyclopropenyl halides, X(-) + (CH)(3)X <= => X(CH)(3) + X(-), are investigated theoretically at four levels of theory, B3LYP/6-311+G**, MP2/6 311+G**, G2(+)MP2//MP2/6-311+G**, and G2(+)//MP2/6-311+G**. Four types of reaction paths, the sigma-attack S(N)2, pi-attack S(N)2'-syn, and S(N)2'-anti and sigmatropic 1,2-shift, are possible for all the halides. In the fluoride anion reactions, two types of stable adducts, syn- and anti-1,2-difluorocyclopropyl anions, can exist on the triple-well-type potential energy surface of the identity substitution reactions with rearrangement of double bond (C=C), S(N)2' syn, and S(N)2'-anti processes. The TSs for the sigma-attack S(N)2 paths have "open" (loose) structures so that the ring positive charges are high rendering strong aromatic cyclopropenyl (delocalized) cation-like character. In contrast, in the pi-attack S(N)2' paths, a lone pair is formed at the unsubstituted carbon (C3), which stabilizes the 1,2-dihalocyclopropyl (delocalized) anion-like TS by two strong n(C)-sigma*(C-F) vicinal charge-transfer delocalization interactions. The barrier height increases in the order S(N)2'-anti < sigma-attack S(N)2 < S(N)2'-syn for X = Cl and Br, whereas for X = F the order is changed to S(N)2' anti < S(N)2'-syn < sigma-attack S(N)2 due to the stable difluoro adduct formation. The sigmatropic 1,2-shift (circumambulatory) reactions have high activation barriers and cannot interfere with the substitution reactions. PMID- 11895418 TI - Cp*(2)TiMe(2): an improved catalyst for the intermolecular addition of n-alkyl- and benzylamines to alkynes. AB - Cp(2)TiMe(2) has been found to be a competent catalyst for the intermolecular addition of sterically less demanding n-alkyl- and benzylamines to internal alkynes. In the presence of 2.0-6.0 mol % of the catalyst, hydroamination reactions between n-propyl-, n-hexyl-, benzyl-, p-methoxybenzyl- or 2 phenylethylamine and diphenylacetylene, 3-hexyne or 4-octyne go to completion within 24 h or less at 114 degrees C (oil bath temperature). After subsequent reduction of the initially formed imines with zinc-modified sodium cyanoborohydride in MeOH at 25 degrees C, the corresponding secondary amines can be isolated in excellent yields (>78%). Hydroamination/reduction sequences employing the unsymmetrically substituted alkyne 1-phenylpropyne give access to mixtures of regioisomeric secondary amines. The observed regioselectivity is low. PMID- 11895419 TI - Accurate calculation of aromaticity of benzene and antiaromaticity of cyclobutadiene: new homodesmotic reactions. AB - Newly designed homodesmotic reactions based on radical systems predict an absolute aromaticity of 29.13 kcal/mol for benzene and an absolute antiaromaticity of 40.28 kcal/mol for cyclobutadiene at the MP4(SDQ)/6-31G(d,p) level. PMID- 11895420 TI - Transmetalation of palladium enolate and its application in palladium-catalyzed homocoupling of alkynes: a room-temperature, highly efficient route to make diynes. AB - A novel pathway for the homocoupling reaction has been achieved using a similar protocol as the cross-coupling reaction. Ethyl bromoacetate is chosen to initiate the coupling reaction through oxidative addition to a Pd(0) species, and an PdBr(enolate) intermediate is formed. This intermediate can undergo double transmetalation with an alkynyl copper reagent, and reductive elimination produces a variety of diynes in high yields. PMID- 11895421 TI - A novel free-radical ring contraction of a cyclic carbamate. AB - During a study on iodocyclocarbamation reactions of 2-styryl-4-piperidones, a novel ring contraction was observed. Iodocyclocarbamation of 2-styryl-4 piperidone 3 gave the bicyclic carbamate 4. Reduction of 4 under free-radical conditions effected a stereoselective ring contraction to provide oxazolidinone 6. A three-electron-three-center mechanism is proposed. PMID- 11895424 TI - [The Gram stain]. PMID- 11895425 TI - [Evaluation of the diagnostic reagents which detect group A Streptococcus with the immunochromatographical method]. AB - Dipstick 'Eiken' Strep A was evaluated for this sensitivity and specificity. Dipstick 'Eiken' Strep A had a capacity to detect Group A Streptococcus in 1.5x10(5) cfu/swab. The sensitivity of Dipstick 'Eiken' Strep A was 4 times higher than the sensitivity of the latex agglutination test (Serodirect 'Eiken' Strep A) and was almost the same as the immunochromatography test (TESTPACK Plus STREP A, CLEARVIEW STREP A and ImmunoCard STAT! STREP A TEST). No cross reaction was observed among 27 strains of 25 species microorganisms with Dipstick 'Eiken' Strep A. Dipstick 'Eiken' Strep A was compared with TESTPACK Plus STREP A among throat swabs from 50 patients with pharyngitis. Dipstick 'Eiken' Strep A had a sensitivity of 92.9%, a specificity of 90.5% and an agreement of 91.8%. Dipstick 'Eiken' Strep A is found to be useful diagnostic assay in the clinical laboratories. PMID- 11895426 TI - [Laboratory-based evaluation of a newly developed immunochromatographic test method by using synthesized oligonucleotide-bound protein probes for simultaneous detection of hepatitis B surface antigen (HBsAg) and specific antibodies to Treponema pallidum]. AB - An immunochromatographic test method using synthesized oligonucleotide-bound protein probes was newly developed and evaluated for simultaneous detection of hepatitis B surface antigen (HBsAg) and specific antibodies to Treponema pallidum (TP). The test principle includes, first, antigen-antibody reaction, and secondly, DNA (oligonucleotide)-DNA interaction. The test device is composed of colloidal gold-labeled HBs antibody and TP antigen, and oligonucleotide-labeled HBs antibody and TP antigen. When the test sample contains HBsAg and/or TP specific antibody, the colloidal gold-labeled probes and oligonucleotide-labeled probes will make a sandwich complex with the target. Then, the formed complex migrates and is immobilized by the respective complementary oligonucleotide fixed on the different lines of the membrane. The color development of colloidal gold was visually read after 20 min and/or 60 min incubation, and easily interpretable, positive or negative. When the performance panels of Boston Biomedica Inc. (BBI) for HBsAg and TP-specific antibody, the results indicated; first, the most positive serum and plasma specimens with 1.2 IU/ml of HBsAg were correctly determined as positive, and secondly, all the test results for TP specific antibody were comparable to the results of fluorescent treponemal antibody test (FTA-ABS). However, when the seroconversion panel of BBI for hepatitis B virus (HBV) infection, the seroconversion was delayed 20 to 30 days when compared to HBV DNA detection. Also, when the clinical serum and plasma samples were tested, sensitivity and specificity were estimated to be 87.0% and 100% for HBsAg, and both 100% for TP-specific antibody, respectively. With these results, we can conclude that this newly developed immunochromato-graphic test method will be applicable to simultaneous detection of multiple antigen and/or specific antibody in a single device, and will be expected to be widely applied in a clinical setting. PMID- 11895427 TI - [Study on the detection methods of Helicobacter pylori from clinical specimens]. AB - We have attempted the detection of Helicobacter pylori using biopsy specimen (tunica mucosa vestibulum ventriculi, tunica mucosa corpus ventriculi) and of Helicobacter pylori Specific Antigen (HpSA) from feces of patients with stomach duodenum disease. During 1991-2000, 753 patients had biopsy test for Gram-stained smear, culture, rapid urease test (RUT), and HpSA by ELISA. As a result, Gram stained smear positive were 609 (44.6%) in 1,357 specimens, culture positive were 984 (72.5%) in 1,357 specimens, and RUT positive were 445 (59.4%) in 749 specimens in method with stomach specimens. 85 (94.4%) in 90 specimens with method of HpSA were positive, and we defined that indirect detection method using feces was most significance compared with other invasive direct method using endoscopy. In addition, antimicrobial susceptibility tests were examined with 5 agents for 349 strains of H. pylori, the MICs of amoxicillin were <0.01-0.19 microgram/ml (MIC80: 0.02 microgram/ml), and clarithromicin were <0.01-50 microgram/ml (MIC80: 0.04 microgram/ml). Amoxicillin resistant strain was not observed. PMID- 11895428 TI - The human beta-globin locus control region. AB - The human beta-globin gene locus is the subject of intense study, and over the past two decades a wealth of information has accumulated on how tissue-specific and stage-specific expression of its genes is achieved. The data are extensive and it would be difficult, if not impossible, to formulate a comprehensive model integrating every aspect of what is currently known. In this review, we introduce the fundamental characteristics of globin locus regulation as well as questions on which much of the current research is predicated. We then outline a hypothesis that encompasses more recent results, focusing on the modification of higher order chromatin structure and recruitment of transcription complexes to the globin locus. The essence of this hypothesis is that the locus control region (LCR) is a genetic entity highly accessible to and capable of recruiting, with great efficiency, chromatin-modifying, coactivator, and transcription complexes. These complexes are used to establish accessible chromatin domains, allowing basal factors to be loaded on to specific globin gene promoters in a developmental stage-specific manner. We conceptually divide this process into four steps: (a) generation of a highly accessible LCR holocomplex; (b) recruitment of transcription and chromatin-modifying complexes to the LCR; (c) establishment of chromatin domains permissive for transcription; (d) transfer of transcription complexes to globin gene promoters. PMID- 11895429 TI - Ferritin from the spleen of the Antarctic teleost Trematomus bernacchii is an M type homopolymer. AB - Ferritin from the spleen of the Antarctic teleost Trematomus bernacchii is composed of a single subunit that contains both the ferroxidase center residues, typical of mammalian H chains, and the carboxylate residues forming the micelle nucleation site, typical of mammalian L chains. Comparison of the amino-acid sequence with those available from lower vertebrates indicates that T. bernacchii ferritin can be classified as an M-type homopolymer. Interestingly, the T. bernacchii ferritin chain shows 85.7% identity with a cold-inducible ferritin chain of the rainbow trout Salmo gairdneri. The structural and functional properties indicate that cold acclimation and functional adaptation to low temperatures are achieved without significant modification of the protein stability. In fact, the stability of T. bernacchii ferritin to denaturation induced by acid or temperature closely resembles that of mesophilic mammalian ferritins. Moreover iron is taken up efficiently and the activation energy of the reaction is 74.9 kJ.mol(-1), a value slightly lower than that measured for the human recombinant H ferritin (80.8 kJ.mol(-1)). PMID- 11895430 TI - Dystrobrevin requires a dystrophin-binding domain to function in Caenorhabditis elegans. AB - Dystrobrevin is one of the intracellular components of the transmembrane dystrophin-glycoprotein complex (DGC). The functional role of this complex in normal and pathological situations has not yet been clearly established. Dystrobrevin disappears from the muscle membrane in Duchenne muscular dystrophy (DMD), which results from dystrophin mutations, as well as in limb girdle muscular dystrophies (LGMD), which results from mutations affecting other members of the DGC complex. These findings therefore suggest that dystrobrevin may play a pivotal role in the progression of these clinically related diseases. In this study, we used the Caenorhabditis elegans model to address the question of the relationship between dystrobrevin binding to dystrophin and dystrobrevin function. Deletions of the dystrobrevin protein were performed and the ability of the mutated forms to bind to dystrophin was tested both in vitro and in a two hybrid assay, as well as their ability to rescue dystrobrevin (dyb-1) mutations in C. elegans. The deletions affecting the second helix of the Dyb-1 coiled-coil domain abolished the binding of dystrobrevin to dystrophin both in vitro and in the two-hybrid assay. These deletions also abolished the rescuing activity of a functional transgene in vivo. These results are consistent with a model according to which dystrobrevin must bind to dystrophin to be able to function properly. PMID- 11895431 TI - Binding of Thermomyces (Humicola) lanuginosa lipase to the mixed micelles of cis parinaric acid/NaTDC. AB - The binding of Thermomyces lanuginosa lipase and its mutants [TLL(S146A), TLL(W89L), TLL(W117F, W221H, W260H)] to the mixed micelles of cis-parinaric acid/sodium taurodeoxycholate at pH 5.0 led to the quenching of the intrinsic tryptophan fluorescence emission (300-380 nm) and to a simultaneous increase in the cis-parinaric acid fluorescence emission (380-500 nm). These findings were used to characterize the Thermomyces lanuginosa lipase/cis-parinaric acid interactions occurring in the presence of sodium taurodeoxycholate. The fluorescence resonance energy transfer and Stern-Volmer quenching constant values obtained were correlated with the accessibility of the tryptophan residues to the cis-parinaric acid and with the lid opening ability of Thermomyces lanuginosa lipase (and its mutants). TLL(S146A) was found to have the highest fluorescence resonance energy transfer. In addition, a TLL(S146A)/oleic acid complex was crystallised and its three-dimensional structure was solved. Surprisingly, two possible binding modes (sn-1 and antisn1) were found to exist between oleic acid and the catalytic cleft of the open conformation of TLL(S146A). Both binding modes involved an interaction with tryptophan 89 of the lipase lid, in agreement with fluorescence resonance energy transfer experiments. As a consequence, we concluded that TLL(S146A) mutant is not an appropriate substitute for the wild type Thermomyces lanuginosa lipase for mimicking the interaction between the wild type enzyme and lipids. PMID- 11895432 TI - High affinity binding between laminin and laminin binding protein of Leishmania is stimulated by zinc and may involve laminin zinc-finger like sequences. AB - In the course of trying to understand the pathogenesis of leishmaniasis in relation to extracellular matrix (ECM) elements, laminin, a major ECM protein, has been found to bind saturably and with high affinity to a 67-kDa cell surface protein of Leishmania donovani. This interaction involves a single class of binding sites, which are ionic in nature, conformation-dependent and possibly involves sulfhydryls. Binding activity was significantly enhanced by Zn2+, an effect possibly mediated through Cys-rich zinc finger-like sequences on laminin. Inhibition studies with monoclonals against polypeptide chains and specific peptides with adhesive properties revealed that the binding site was localized in one of the nested zinc finger consensus sequences of B1 chain containing the specific pentapeptide sequence, YIGSR. Furthermore, incubation of L. donovani promastigotes with C(YIGSR)3-NH2 peptide amide or antibody directed against the 67-kDa laminin-binding protein (LBP) induced tyrosine phosphorylation of proteins with a molecular mass ranging from 115 to 130 kDa. These studies suggest a role for LBP in the interaction of parasites with ECM elements, which may mediate one or more downstream signalling events necessary for establishment of infection. PMID- 11895433 TI - Identification of residues critical for activity of the wound-induced leucine aminopeptidase (LAP-A) of tomato. AB - The importance of two putative Zn2+-binding (Asp347, Glu429) and two catalytic (Arg431, Lys354) residues in the tomato leucine aminopeptidase (LAP-A) function was tested. The impact of substitutions at these positions, corresponding to the bovine LAP residues Asp255, Glu334, Arg336, and Lys262, was evaluated in His6-LAP A fusion proteins expressed in Escherichia coli. Sixty-five percent of the mutant His6-LAP-A proteins were unstable or had complete or partial defects in hexamer assembly or stability. The activity of hexameric His6-LAP-As on Xaa-Leu and Leu Xaa dipeptides was tested. Most substitutions of Lys354 (a catalytic residue) resulted in His6-LAP-As that cleaved dipeptides at slower rates. The Glu429 mutants (a Zn2+-binding residue) had more diverse phenotypes. Some mutations abolished activity and others retained partial or complete activity. The E429D His6-LAP-A enzyme had Km and kcat values similar to the wild-type His6-LAP-A. One catalytic (Arg431) and one Zn-binding (Asp347) residue were essential for His6 LAP-A activity, as most R431 and D347 mutant His6-LAP-As did not hydrolyze dipeptides. The R431K His6-LAP-A that retained the positive charge had partial activity as reflected in the 4.8-fold decrease in kcat. Surprisingly, while the D347E mutant (that retained a negative charge at position 347) was inactive, the D347R mutant that introduced a positive charge retained partial activity. A model to explain these data is proposed. PMID- 11895434 TI - Characterization of the self-splicing products of two complex Naegleria LSU rDNA group I introns containing homing endonuclease genes. AB - The two group I introns Nae.L1926 and Nmo.L2563, found at two different sites in nuclear LSU rRNA genes of Naegleria amoebo-flagellates, have been characterized in vitro. Their structural organization is related to that of the mobile Physarum intron Ppo.L1925 (PpLSU3) with ORFs extending the L1-loop of a typical group IC1 ribozyme. Nae.L1926, Nmo.L2563 and Ppo.L1925 RNAs all self-splice in vitro, generating ligated exons and full-length intron circles as well as internal processed excised intron RNAs. Formation of full-length intron circles is found to be a general feature in RNA processing of ORF-containing nuclear group I introns. Both Naegleria LSU rDNA introns contain a conserved polyadenylation signal at exactly the same position in the 3' end of the ORFs close to the internal processing sites, indicating an RNA polymerase II-like expression pathway of intron proteins in vivo. The intron proteins I-NaeI and I-NmoI encoded by Nae.L1926 and Nmo.L2563, respectively, correspond to His-Cys homing endonucleases of 148 and 175 amino acids. I-NaeI contains an additional sequence motif homologous to the unusual DNA binding motif of three antiparallel beta sheets found in the I-PpoI endonuclease, the product of the Ppo.L1925 intron ORF. PMID- 11895435 TI - The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus: a comparative study on the redox properties of the metal clusters present in the dinitrogenase components. AB - The dinitrogenase component proteins of the conventional Mo nitrogenase (MoFe protein) and of the alternative Fe-only nitrogenase (FeFe protein) were both isolated and purified from Rhodobacter capsulatus, redox-titrated according to the same procedures and subjected to an EPR spectroscopic comparison. In the course of an oxidative titration of the MoFe protein (Rc1Mo) three significant S = 1/2 EPR signals deriving from oxidized states of the P-cluster were detected: (1) a rhombic signal (g = 2.07, 1.96 and 1.83), which showed a bell-shaped redox curve with midpoint potentials (Em) of -195 mV (appearance) and -30 mV (disappearance), (2) an axial signal (g(parallel) = 2.00, g perpendicular = 1.90) with almost identical redox properties and (3) a second rhombic signal (g = 2.03, 2.00, 1.90) at higher redox potentials (> 100 mV). While the 'low-potential' rhombic signal and the axial signal have been both attributed to the one-electron oxidized P-cluster (P1+) present in two conformationally different proteins, the 'high-potential' rhombic signal has been suggested rather to derive from the P3+ state. Upon oxidation, the FeFe protein (Rc1Fe) exhibited three significant S = 1/2 EPR signals as well. However, the Rc1Fe signals strongly deviated from the MoFe protein signals, suggesting that they cannot simply be assigned to different P-cluster states. (a) The most prominent feature is an unusually broad signal at g = 2.27 and 2.06, which proved to be fully reversible and to correlate with catalytic activity. The cluster giving rise to this signal appears to be involved in the transfer of two electrons. The midpoint potentials determined were: -80 mV (appearance) and 70 mV (disappearance). (b) Under weakly acidic conditions (pH 6.4) a slightly altered EPR signal occurred. It was characterized by a shift of the g values to 2.22 and 2.05 and by the appearance of an additional negative absorption-shaped peak at g = 1.86. (c) A very narrow rhombic EPR signal at g = 2.00, 1.98 and 1.96 appeared at positive redox potentials (Em = 80 mV, intensity maximum at 160 mV). Another novel S = 1/2 signal at g = 1.96, 1.92 and 1.77 was observed on further, enzymatic reduction of the dithionite-reduced state of Rc1Fe with the dinitrogenase reductase component (Rc2Fe) of the same enzyme system (turnover conditions in the presence of N2 and ATP). When the Rc1Mo protein was treated analogously, neither this 'turnover signal' nor any other S = 1/2 signal were detectable. All Rc1Fe-specific EPR signals detected are discussed and tentatively assigned with special consideration of the reference spectra obtained from Rc1Mo preparations. PMID- 11895436 TI - DNA supercoiling in Escherichia coli is under tight and subtle homeostatic control, involving gene-expression and metabolic regulation of both topoisomerase I and DNA gyrase. AB - DNA of prokaryotes is in a nonequilibrium structural state, characterized as 'active' DNA supercoiling. Alterations in this state affect many life processes and a homeostatic control of DNA supercoiling has been suggested [Menzel, R. & Gellert, M. (1983) Cell 34, 105-113]. We here report on a new method for quantifying homeostatic control of the high-energy state of in vivo DNA. The method involves making small perturbation in the expression of topoisomerase I, and measuring the effect on DNA supercoiling of a reporter plasmid and on the expression of DNA gyrase. In a separate set of experiments the expression of DNA gyrase was manipulated and the control on DNA supercoiling and topoisomerase I expression was measured [part of these latter experiments has been published in Jensen, P.R., van der Weijden, C.C., Jensen, L.B., Westerhoff, H.V. & Snoep, J.L. (1999) Eur. J. Biochem. 266, 865-877]. Of the two regulatory mechanisms via which homeostasis is conferred, regulation of enzyme activity or regulation of enzyme expression, we quantified the first to be responsible for 72% and the latter for 28%. The gene expression regulation could be dissected to DNA gyrase (21%) and to topoisomerase I (7%). On a scale from 0 (no homeostatic control) to 1 (full homeostatic control) we quantified the homeostatic control of DNA supercoiling at 0.87. A 10% manipulation of either topoisomerase I or DNA gyrase activity results in a 1.3% change of DNA supercoiling only. We conclude that the homeostatic regulation of the nonequilibrium DNA structure in wild-type Escherichia coli is almost complete and subtle (i.e. involving at least three regulatory mechanisms). PMID- 11895437 TI - Structural basis for the inhibitory efficacy of efavirenz (DMP-266), MSC194 and PNU142721 towards the HIV-1 RT K103N mutant. AB - The K103N substitution is a frequently observed HIV-1 RT mutation in patients who do not respond to combination-therapy. The drugs Efavirenz, MSC194 and PNU142721 belong to the recent generation of NNRTIs characterized by an improved resistance profile to the most common single point mutations within HIV-1 RT, including the K103N mutation. In the present study we present structural observations from Efavirenz in complex with wild-type protein and the K103N mutant and PNU142721 and MSC194 in complex with the K103N mutant. The structures unanimously indicate that the K103N substitution induces only minor positional adjustments of the three inhibitors and the residues lining the binding pocket. Thus, compared to the corresponding wild-type structures, these inhibitors bind to the mutant in a conservative mode rather than through major rearrangements. The structures implicate that the reduced inhibitory efficacy should be attributed to the changes in the chemical environment in the vicinity of the substituted N103 residue. This is supported by changes in hydrophobic and electrostatic interactions to the inhibitors between wild-type and K103N mutant complexes. These potent inhibitors accommodate to the K103N mutation by forming new interactions to the N103 side chain. Our results are consistent with the proposal by Hsiou et al. [Hsiou, Y., Ding, J., Das, K., Clark, A.D. Jr, Boyer, P.L., Lewi, P., Janssen, P.A., Kleim, J.P., Rosner, M., Hughes, S.H. & Arnold, E. (2001) J. Mol. Biol. 309, 437-445] that inhibitors with good activity against the K103N mutant would be expected to have favorable interactions with the mutant asparagines side chain, thereby compensating for resistance caused by stabilization of the mutant enzyme due to a hydrogen-bond network involving the N103 and Y188 side chains. PMID- 11895438 TI - Increase of the deacylation rate of PBP2x from Streptococcus pneumoniae by single point mutations mimicking the class A beta-lactamases. AB - The class A beta-lactamases and the transpeptidase domain of the penicillin binding proteins (PBPs) share the same topology and conserved active-site residues. They both react with beta-lactams to form acylenzymes. The stability of the PBP acylenzymes results in the inhibition of the transpeptidase function and the antibiotic activity of the beta-lactams. In contrast, the deacylation of the beta-lactamases is extremely fast, resulting in a high turnover of beta-lactam hydrolysis, which confers resistance to these antibiotics. In TEM-1 beta lactamase from Escherichia coli, Glu166 is required for the fast deacylation and occupies the same spatial location as Phe450 in PBP2x from Streptococcus pneumoniae. To gain insight into the deacylation mechanism of both enzymes, Phe450 of PBP2x was replaced by various residues. The introduction of ionizable side chains increased the deacylation rate, in a pH-dependent manner, for the acidic residues. The aspartic acid-containing variant had a 110-fold faster deacylation at pH 8. The magnitude of this effect is similar to that observed in a naturally occurring variant of PBP2x, which confers increased resistance to cephalosporins. PMID- 11895439 TI - Binding of N-acetyl-N '-beta-D-glucopyranosyl urea and N-benzoyl-N '-beta-D glucopyranosyl urea to glycogen phosphorylase b: kinetic and crystallographic studies. AB - Two substituted ureas of beta-D-glucose, N-acetyl-N'-beta-D-glucopyranosyl urea (Acurea) and N-benzoyl-N'-beta-D-glucopyranosyl urea (Bzurea), have been identified as inhibitors of glycogen phosphorylase, a potential target for therapeutic intervention in type 2 diabetes. To elucidate the structural basis of inhibition, we determined the structure of muscle glycogen phosphorylase b (GPb) complexed with the two compounds at 2.0 A and 1.8 A resolution, respectively. The structure of the GPb-Acurea complex reveals that the inhibitor can be accommodated in the catalytic site of T-state GPb with very little change in the tertiary structure. The glucopyranose moiety makes the standard hydrogen bonds and van der Waals contacts as observed in the GPb-glucose complex, while the acetyl urea moiety is in a favourable electrostatic environment and makes additional polar contacts with the protein. The structure of the GPb-Bzurea complex shows that Bzurea binds tightly at the catalytic site and induces substantial conformational changes in the vicinity of the catalytic site. In particular, the loop of the polypeptide chain containing residues 282-287 shifts 1.3-3.7 A (Calpha atoms) to accommodate Bzurea. Bzurea can also occupy the new allosteric site, some 33 A from the catalytic site, which is currently the target for the design of antidiabetic drugs. PMID- 11895440 TI - Involvement of GFA1, which encodes glutamine-fructose-6-phosphate amidotransferase, in the activation of the chitin synthesis pathway in response to cell-wall defects in Saccharomyces cerevisiae. AB - Cell-wall damage caused by mutations of cell-wall-related genes triggers a compensatory mechanism which eventually results in hyperaccumulation of chitin reaching 20% of the cell-wall dry mass. We show that activation of chitin synthesis is accompanied by a rise, from 1.3-fold to 3.5-fold according to the gene mutation, in the expression of most of the genes encoding enzymes of the chitin metabolic pathways. Evidence that GFA1, which encodes glutamine-fructose-6 Phosphate amidotransferase (Gfa1p), the first committed enzyme of this pathway, plays a major role in this process was as follows. Activation of chitin synthesis in the cell-wall mutants correlated with activation of GFA1 and with a proportional increase in Gfa1p activity. Overexpression of GFA1 caused an approximately threefold increase in chitin in the transformed cells, whereas chitin content was barely affected by the joint overexpression of CHS3 and CHS7. Introduction of a gfa1-97 allele mutation in the cell-wall-defective gas1Delta mutant or cultivation of this mutant in a hyperosmotic medium resulted in reduction in chitin synthesis that was proportional to the decrease in Gfa1p activity. Finally, the stimulation of chitin production was also accompanied by an increase in pools of fructose 6-Phosphate, a substrate of Gfa1p. In quantitative terms, we estimated the flux-coefficient control of Gfa1p to be in the range of 0.90, and found that regulation of the chitin metabolic pathway was mainly hierarchical, i.e. dominated by regulation of the amount of newly synthesized GFA1 protein. In the search for the mechanism by which GFA1 is activated in response to cell-wall perturbations, we could only show that neither MCM1 nor RLM1, which encode two transcriptional factors of the MADS box family that are required for expression of cell-cycle and cell-wall-related genes, was involved in this process. PMID- 11895441 TI - Construction and characterization of trifunctional single-chain urokinase-type plasminogen activators. AB - Two chimeric proteins have been constructed. One consists of four parts: a portion of the low molecular mass single-chain urokinase-type plasminogen activator (scu-PA-32K, residues 144-411), a 15-mer linker sequence, the C terminal amino-acid sequence (residues 53-65) of hirudin (Hir), and an RGD sequence derived from the leech protein decorsin, i.e. scu-PA(32 k)-linker-Hir (residues 53-65)-RGD peptide. The other comprises two main segments: scu-PA(32 k) and hirudin into which RGDSP is inserted between its residues 33 and 34, i.e. hirudin (residues 1-33)-RGDSP-hirudin (residues 34-65)-scu-PA(32 k). These two chimeric genes were expressed in Escherichia coli, and the products were purified by Zn2+-chelating Sepharose 4B chromatography and benzamidine Sepharose 6B chromatography. Our results suggested that these two chimeric proteins not only had plasminogen-dependent fibrinolytic activity, but also possessed platelet aggregation inhibitory activity and antithrombin activity. PMID- 11895442 TI - Investigation of a functional requirement for isoprenylation by the human prostacyclin receptor. AB - In the current study, we have established that the human (h) prostacyclin receptor (IP) is isoprenylated in whole cells. Through site directed mutagenesis and generation of the isoprenylation defective hIPSSLC, it was established that while isoprenylation of hIP does not influence ligand binding, it is obligatory for agonist activation of adenylyl cyclase and cAMP generation. Overexpression of GalphaS significantly augmented cAMP generation by the hIP but not by the hIPSSLC. Moreover, GalphaS co-immunoprecipitated with hIP following agonist activation but did not co-immunoprecipitate with hIPSSLC. Whereas hIP mediated concentration-dependent activation of phospholipase C (PLC); the extent of PLC activation by hIPSSLC was impaired compared to hIP. Co-expression of Galphaq significantly augmentated intracellular calcium mobilization by the hIP but not by hIPSSLC. Moreover, whereas Galphaq co-immunoprecipitated with hIP, it failed to co-immunoprecipitate with hIPSSLC. While both the hIP and hIPSSLC underwent agonist-induced internalization, the kinetics and extent of hIPSSLC internalization was impaired compared to hIP. Altering the CAAX motif of the hIP from a farnesyl (-CSLC) to a geranylgeranyl (-CSLL) isoprene acceptor, to generate hIPCSLL, did not affect ligand binding and yielded a receptor that exhibited identical signalling through both Gs- and Gq-coupled effectors to that of hIP. Thus, whereas isoprenylation of hIP does not influence ligand binding, it is functionally imperative in regulating post-receptor events including agonist activation of adenylyl cyclase, for efficient activation of PLC and for receptor internalization. Though the nature of the isoprenoid attached to hIP does not act as a major determinant, the presence of an isoprenoid group, for example farnesyl or geranylgeranyl, is required for functional receptor-G protein interaction and coupling and for efficient agonist- induced receptor internalization. PMID- 11895443 TI - Drug recognition of a DNA single strand break: nogalamycin intercalation between coaxially stacked hairpins. AB - Two DNA hairpin motifs (5'-GCGAAGC-3' and 5'-ACGA AGT-3'), both stabilized by a 5'-GAA loop, have been used to design novel intramolecular double hairpin structures (5'-GCGAAGCACGAAGT-3' and 5'-ACGAAGTGCG AAGC-3') in which coaxial stacking of the two hairpin components generates a double-stranded stem region effectively with a single-strand break in the middle of the sequence at either the TG or CA step between unconnected 3' and 5' terminal bases. We have investigated by NMR the conformation and dynamics of the DNA at the strand break site. We show that mutual stacking significantly enhances the stability of each hairpin. Further, the anthracycline antibiotic nogalamycin binds cleanly to the 5'-TG (5'-CA) site formed by the mutually stacked hairpins despite the break in the sugar-phosphate backbone on one strand. The complex resembles the structure of nogalamycin-DNA complexes with the drug bound at 5'-TG sites in intact duplex sequences, with pi-stacking interactions probably the single dominant stabilizing interaction. PMID- 11895444 TI - The lipase/acyltransferase from Candida parapsilosis: molecular cloning and characterization of purified recombinant enzymes. AB - Candida parapsilosis has been previously shown to produce a lipase (i.e. able to catalyze efficiently the hydrolysis of insoluble lipid esters such as triacylglycerols) that preferentially catalyses transfer reactions such as alcoholysis in the presence of suitable nucleophiles other than water, even in aqueous media with high (> 0.9) water thermodynamic activity. The present work describes the cloning and the overexpression of the gene coding for this enzyme. Two ORFs (CpLIP1 and CpLIP2) were isolated. The deduced 465-amino-acid protein sequences contained the consensus motif (G-X-S-X-G) which is conserved among lipolytic enzymes. Only one of the two deduced proteins (CpLIP2) contained peptide sequences obtained from the purified lipase/acyltransferase. Homology investigations showed that CpLIP2 has similarities principally with 11 lipases produced by C. albicans (42-61%) and the lipase A from Candida antarctica (31%) but not with the other lipases sequenced so far. Both CpLIP1 and CpLIP2 were expressed in Saccharomyces cerevisiae, but only CpLIP2 coded for an active protein. The substrate specificity and the catalytic behavior of purified recombinant CpLIP2, with or without a C-terminal histidine tag, were not changed compared to those of the native lipase. PMID- 11895445 TI - Lipopolysaccharide regions involved in the activation of Escherichia coli outer membrane protease OmpT. AB - OmpT is an integral outer membrane protease of Escherichia coli. Overexpression of OmpT in E. coli and subsequent in vitro folding of the produced inclusion bodies yielded protein with a native-like structure. However, enzymatically active protease was only obtained after addition of the outer membrane lipid lipopolysaccharide (LPS). OmpT is the first example of an enzyme that requires LPS for activity. In this study, we investigated the nature of this activation. Circular dichroism analysis showed that binding of LPS did not lead to large structural changes. Titration of OmpT with LPS and determining the resulting OmpT activity with a fluorimetric assay yielded a dissociation constant of 10-4 m for E. coli K-12 LPS. Determining the dissociation constants for different LPS chemotypes revealed that a fully acylated lipid A part is minimally required for activation of OmpT. The heptose-bound phosphates in the inner core region were also important for activation. The affinity for LPS was not dependent on the concentration of substrate, neither was affinity for the substrate influenced by the concentration of LPS. This indicated that LPS most likely does not act at the level of substrate binding. We hypothesize that LPS induces a subtle conformational change in the protein that is required for obtaining a native active site geometry. PMID- 11895446 TI - Cloning and expression in Pichia pastoris of a blue mussel (Mytilus edulis) beta mannanase gene. AB - Using PCR, cloning and sequencing techniques, a 1.1-kb complementary DNA fragment encoding for a beta-mannanase (mannan endo-1,4-beta-mannosidase, EC 3.2.1.78) has been identified in the digestive gland of blue mussel, Mytilus edulis. The cDNA sequence shows significant sequence identity to several beta-mannanases in glycoside hydrolase family 5. The beta-mannanase gene has been isolated and sequenced from gill tissue of blue mussel and contains five introns. The beta mannanase has been expressed extracellularly in Pichia pastoris using the Saccharomyces cerevisiae alpha-factor signal sequence. The beta-mannanase was produced in a 14-L fermenter with an expression level of 900 mg.L-1. The expression level is strongly affected by the induction temperature. A two-step purification procedure, composed of a combination of immobilized metal ion affinity chromatography and ion exchange chromatography, is required to give a pure beta-mannanase. However, due to post-translational modifications, structural varieties regarding molecular mass and isoelectric point were obtained. The specific activity of the purified recombinant M. edulis beta-mannanase was close to that of the wild-type enzyme. Also pH and temperature optima were the same as for the native protein. In conclusion, P. pastoris is regarded as a suitable host strain for the production of blue mussel beta-mannanase. This is the first time a mollusc beta-mannanase has been characterized at the DNA level. PMID- 11895447 TI - Differential effects of ergosterol and cholesterol on Cdk1 activation and SRE driven transcription. AB - Cholesterol is essential for cell growth and division, but whether this is just a consequence of its use in membrane formation or whether it also elicits regulatory actions in cell cycle machinery remains to be established. Here, we report on the specificity of this action of cholesterol in human cells by comparing its effects with those of ergosterol, a yeast sterol structurally similar to cholesterol. Inhibition of cholesterol synthesis by means of SKF 104976 in cells incubated in a cholesterol-free medium resulted in cell proliferation inhibition and cell cycle arrest at G2/M phase. These effects were abrogated by cholesterol added to the medium but not by ergosterol, despite that the latter was used by human cells and exerted similar homeostatic actions, as the regulation of the transcription of an SRE-driven gene construct. In contrast to cholesterol, ergosterol was unable to induce cyclin B1 expression, to activate Cdk1 and to resume cell cycle in cells previously arrested at G2. This lack of effect was not due to cytotoxicity, as cells exposed to ergosterol remained viable and, upon supplementing with UCN-01, an activator of Cdk1, they progressed through mitosis. However, in the presence of suboptimal concentrations of cholesterol, ergosterol exerted synergistic effects on cell proliferation. This is interpreted on the basis of the differential action of these sterols, ergosterol contributing to cell membrane formation and cholesterol being required for Cdk1 activation. In summary, the action of cholesterol on G2 traversal is highly specific and exerted through a mechanism different to that used for cholesterol homeostasis, reinforcing the concept that cholesterol is a specific regulator of cell cycle progression in human cells. PMID- 11895448 TI - Cloning and expression of cDNA coding for bouganin. AB - Bouganin is a ribosome-inactivating protein that recently was isolated from Bougainvillea spectabilis Willd. In this work, the cloning and expression of the cDNA encoding for bouganin is described. From the cDNA, the amino-acid sequence was deduced, which correlated with the primary sequence data obtained by amino acid sequencing on the native protein. Bouganin is synthesized as a pro-peptide consisting of 305 amino acids, the first 26 of which act as a leader signal while the 29 C-terminal amino acids are cleaved during processing of the molecule. The mature protein consists of 250 amino acids. Using the cDNA sequence encoding the mature protein of 250 amino acids, a recombinant protein was expressed, purified and characterized. The recombinant molecule had similar activity in a cell-free protein synthesis assay and had comparable toxicity on living cells as compared to the isolated native bouganin. PMID- 11895449 TI - Characterization of RNA polymerase III transcription factor TFIIIC from the mulberry silkworm, Bombyx mori. AB - Fractionation of nuclear extracts from posterior silk glands of mulberry silkworm Bombyx mori, resolved the transcription factor TFIIIC into two components (designated here as TFIIIC and TFIIIC1) as in HeLa cell nuclear extracts. The reconstituted transcription of tRNA genes required the presence of both components. The affinity purified TFIIIC is a heteromeric complex comprising of five subunits ranging from 44 to 240 kDa. Of these, the 51-kDa subunit could be specifically crosslinked to the B box of tRNA1Gly. Purified swTFIIIC binds to the B box sequences with an affinity in the same range as of yTFIIIC or hTFIIIC2. Although an histone acetyl transferase (HAT) activity was associated with the TFIIIC fractions during the initial stages of purification, the HAT activity, unlike the human TFIIIC preparations, was separated at the final DNA affinity step. The tRNA transcription from DNA template was independent of HAT activity but the repressed transcription from chromatin template could be partially restored by external supplementation of the dissociated HAT activity. This is the first report on the purification and characterization of TFIIIC from insect systems. PMID- 11895450 TI - Structures of two histidine ammonia-lyase modifications and implications for the catalytic mechanism. AB - Histidine ammonia-lyase (EC 4.3.1.3) catalyzes the nonoxidative elimination of the alpha-amino group of histidine using a 4-methylidene-imidazole-5-one (MIO), which is formed autocatalytically from the internal peptide segment 142Ala-Ser Gly. The structure of the enzyme inhibited by a reaction with l-cysteine was established at the very high resolution of 1.0 A. Five active center mutants were produced and their catalytic activities were measured. Among them, mutant Tyr280- >Phe could be crystallized and its structure could be determined at 1.7 A resolution. It contains a planar sp2-hybridized 144-N atom of MIO, in contrast to the pyramidal sp3-hybridized 144-N of the wild-type. With the planar 144-N atom, MIO assumes the conformation of a putative intermediate aromatic state of the reaction, demonstrating that the conformational barrier between aromatic and wild type states is very low. The data led to a new proposal for the geometry for the catalyzed reaction, which also applies to the closely related phenylalanine ammonia-lyase (EC 4.3.1.5). Moreover, it suggested an intermediate binding site for the released ammonia. PMID- 11895451 TI - A proposal to redefine familial combined hyperlipidaemia -- third workshop on FCHL held in Barcelona from 3 to 5 May 2001, during the scientific sessions of the European Society for Clinical Investigation. PMID- 11895452 TI - Hyperprolactinaemia in patients with heart failure: clinical and immunogenetic correlations. AB - BACKGROUND: Prolactin represents a stimulatory link between the neuroendocrine and immune systems, but its involvement in the neurohumoral adaptations to heart failure (HF) has not been explored. METHODS: We prospectively studied 55 patients (45 males, 10 females, age 48 +/- 7 years) with NYHA Class II/III HF due either to dilated cardiomyopathy (CMP) (n = 33) or ischemic heart disease (IHD) (n = 22). Serum prolactin levels were determined by radioimmunoassay, soluble interleukin-2 receptor (sIL-2R) levels by enzyme-linked immunoassay and HLA-DQ genotyping with PCR. Left ventricular ejection fraction (LVEF) and end-diastolic diameter (LVEDd) were assessed echocardiographically. RESULTS: Hyperprolactinaemia (17.3 +/- 4 ng mL-1 [Group I] vs. 4.64 +/- 2 ng mL-1 [Group II], P < 0.01) was found in 14 patients (8 with IHD, 6 with CMP). The distribution of HLA-DQB1 alleles was compared in the two groups and showed a significant increase in the frequency of *0301 (86% in Group I vs. 45% in Group II, P < 0.05). Histidine at position 30 of the HLA-DQB1 gene was found in 22% of Group II but in none of Group I patients. Furthermore, there was an inverse correlation between the presence of histidine at position 30 and the levels of serum prolactin. Both sIL-2R levels, a marker of T-cell activation, and concanavalin A-stimulated lymphocyte proliferation were lower in Group I patients (561 +/- 106 vs. 804 +/- 109 pg mL-1 and 20.8 +/- 4 vs. 37.3 +/- 5 cpmX103 [3H] thymidine, respectively). LVEF was significantly higher (32 +/- 5%) and LVEDd smaller (62.0 +/- 6 mm) in Group I compared to Group II (25 +/- 4% and 68.0 +/- 5 mm, respectively, P < 0.01) patients. CONCLUSION: Hyperprolactinaemia presents in 25% of patients with HF and may reflect decreased activation of T-lymphocytes associated with relatively preserved LV systolic function which is under immune genetic control at the HLA-DQ locus. PMID- 11895454 TI - Insulin resistance affects the regulation of lipoprotein lipase in the postprandial period and in an adipose tissue-specific manner. AB - AIMS: Insulin is a potent stimulator of adipose tissue lipoprotein lipase (LPL). Logically, the postprandial period is therefore a privileged time of the day for the regulation of LPL by insulin in this tissue. It is not clear to what extent a defect such as insulin resistance could affect this regulation and contribute to postprandial, as well as fasting, hypertriglyceridaemia. The aim of the present protocol was to study the relationship between insulin resistance and LPL in adipose tissue and in plasma, in the particular context of the postprandial period. METHODS: For this study, 26 adult nondiabetic individuals (12 women and 14 men) with a wide range of whole-body insulin-mediated glucose uptake (as assessed with an insulin suppression test) were studied. An abdominal subcutaneous fat biopsy on one occasion, and post-heparin plasma on another occasion, were obtained 4 h into a standardized meal profile administered in the fasting state. RESULTS: Postprandial triglyceride excursions (evaluated by the incremental area under the curve during the metabolic meal profile) were inversely correlated to adipose tissue LPL mRNA levels (rho = -0.43, P < 0.03) as well as to adipose tissue LPL heparin-releasable activity (rho = -0.58, P < 0.01). Steady-state plasma glucose (SSPG) concentrations during the insulin suppression test, a reflection of the degree of insulin resistance, were also negatively correlated to adipose tissue LPL mRNA (rho = -0.50, P < 0.02) and activity (rho = -0.56, P < 0.01). There was no correlation between plasma post heparin LPL activity/mass and postprandial triglycerides nor with insulin resistance. CONCLUSION: Regulation of adipose tissue LPL is significantly affected in insulin-resistant individuals in the postprandial period. This presumed impaired effect of insulin on LPL postprandially could be an important contributor to the atherogenic dyslipidaemia described in insulin resistance syndrome. PMID- 11895453 TI - Dietary epoxy fatty acids are absorbed in healthy women. AB - BACKGROUND: Epoxy fats in the diet may adversely affect human health. There are no data on the absorption of these fats in humans. METHODS: Triglycerides were synthesized containing two U-13C-labelled monoepoxy or diepoxy stearic acid molecules. Apparently healthy women consumed a standardized fatty meal (30 g fat) containing either 20 mg monoepoxy or 25 mg diepoxy fat (n = 6 and n = 7, respectively). Plasma lipid [U-13C]monoepoxy and diepoxy stearate concentrations were determined (0-24 h) by gas chromatography-mass spectrometry. RESULTS: Plasma triglycerides increased from 1.05 +/- 0.12 to 1.83 +/- 0.13 mmol L-1 (n = 6) and from 1.10 +/- 0.19 to 1.41 +/- 0.27 mmol L-1 (n = 7) (both P < 0.001). Plasma [U 13C]monoepoxy and diepoxy stearate levels increased to 0.18 +/- 0.07 micromol L-1 (n = 6) and to 0.08 +/- 0.03 micromol L-1 (n = 7), respectively. Monoepoxy triglyceride was better absorbed than diepoxy triglyceride: 17 +/- 4 vs. 8 +/- 1% of dose (determined from area under curve (plasma 13C) normalized to that of absorbed triglycerides (plasma 12C); P < 0.02 after log transformation). The absorption of monoepoxy- and diepoxy-labelled triglycerides was related to that of normal triglycerides (r = 0.80, P < 0.05 and r = 0.91, P < 0.001, respectively). CONCLUSIONS: Monoepoxy fats are better absorbed than diepoxy fats in women (17 +/- 4 vs. 8 +/- 1% of dose, P = 0.02). This difference in absorption is important when considering the relative toxicity of epoxidized material in the food chain. PMID- 11895455 TI - Sodium-lithium countertransport is increased in normoalbuminuric type 1 diabetes but is not related to other risk factors for microangiopathy. AB - BACKGROUND: It has been reported that sodium-lithium countertransport (Na/Li CT) activity is increased in patients with diabetes mellitus and that this increased Na/Li CT activity is associated with the development of diabetic nephropathy. It is unclear however, whether Na/Li CT is related to other pathophysiological factors in diabetic patients. We studied kinetic parameters of Na/Li CT activity together with other putative risk factors for microangiopathy in normoalbuminuric type 1 diabetic patients and matched control subjects. SUBJECTS AND METHODS: We measured maximum velocity (Vmax) and sodium affinity (Km) of Na/Li CT in 53 diabetic patients and 45 healthy controls. Endothelial function was assessed by monitoring forearm vascular response to intrabrachial infusion of acetylcholine. Blood samples were collected for measurement of HbA1c, glucose, insulin and lipids. Blood pressure was measured intra-arterially. Renal haemodynamics were measured by inulin/p-aminohippurate clearance. Urinary albumin was measured by enzyme-linked immunosorbent assay. Transcapillary escape of albumin (TERalb) was calculated by the disappearance curve of 125I-labelled albumin. RESULTS: Vmax was increased in diabetic patients (779 +/- 36 micromol Li+ h-1 L-1 erythrocytes vs. 623 +/- 35 in controls, P < 0.01), whereas Km was decreased (64 +/- 16 mmol L-1 vs. 76 +/- 27 in controls, P = 0.03). The ratio of Vmax : Km was 12.4 +/- 0.6 in diabetic patients and 8.9 +/- 0.9 in controls (P < 0.001). When comparing diabetic patients in the lowest and highest quartile of Vmax or Km there were no differences in blood pressure, renal haemodynamics, urinary albumin excretion, TERalb, endothelial function, HbA1c, glucose, insulin, or lipid profile. CONCLUSION: Na/Li CT is increased in uncomplicated type 1 diabetes and characterized by an increase in Vmax and a decrease in Km. The increase in Na/Li CT is not associated with changes in endothelial function, degree of metabolic control, blood pressure or renal haemodynamics. PMID- 11895457 TI - Serum angiogenin concentrations in young patients with diabetes mellitus. AB - BACKGROUND: Angiogenin serum levels were measured in a large group of type 1 diabetic young patients, looking at whether increased Angiogenin concentrations are associated with long-term glycemic control and microvascular complications. MATERIALS AND METHODS: Four groups of patients were compared to 223 age- and sex- matched healthy controls: 196 type 1 diabetic patients (age range 3-24 years, onset of diabetes before the age of 12 years; duration of disease longer than 2 years), without microvascular complications were divided into three groups on the basis of age (group 1, n = 37, age < 6 years; group 2, n = 71, age 6-12 years; group 3, n = 88, age > 12 years); 53 adolescents and young adults (age 16.1-29.7 years) with diabetic microvascular complications (background, preproliferative or proliferative retinopathy, albumin excretion rate 20-200 microg min-1) (group 4). RESULTS: Angiogenin serum levels were significantly increased in diabetic pre school and pre-pubertal children, and particularly elevated in pubertal subjects compared with age- and sex-matched controls. Adolescents and young adults with microvascular complications had very high angiogenin concentrations. One-year mean HbA1c values were correlated with angiogenin levels (r = 0.389; p < 0.01). In poorly controlled diabetics (HbA1c > 10%), long-term (2 years) improvement of glycemic control determined a significant reduction of angiogenin concentrations in both pre-school and pre-pubertal children as well as in pubertal youngsters. CONCLUSIONS: Angiogenin serum concentrations are increased in diabetic children even before puberty. Severity of microvascular complications is associated with markedly increased angiogenin serum levels. Long-term tight glycemic control determines a consistent reduction of angiogenin concentrations. PMID- 11895456 TI - A small reduction in glomerular filtration is accompanied by insulin resistance in type I diabetes patients with diabetic nephrophathy. AB - BACKGROUND: Insulin sensitivity and insulin clearance are compromised in end stage renal disease but it has not been fully established whether they are altered in earlier stages of diabetic nephropathy. DESIGN: We studied three groups of patients with type 1 diabetes; 10 with no sign of nephropathy, 11 with albuminuria (> 20 microg min-1) but normal glomerular filtration rate (GFR) and eight with a small reduction in GFR, (43-73 mL min-1 1.73 m-2). The groups were matched for age (range 36-61 years), body mass index (BMI), diabetes duration and glycaemic control. The euglycaemic hyperinsulinaemic clamp technique was utilized to study insulin sensitivity (M-value) and metabolic clearance rate for insulin. Needle biopsies from abdominal subcutaneous fat tissue were obtained to study insulin binding, insulin degradation, insulin-stimulated glucose uptake and anti lipolysis in adipocytes in vitro. RESULTS: Patients with reduced GFR were more insulin-resistant (M-value 5.7 +/- 0.7 mg kg LBM-1 min-1) than those without nephropathy (9.6 +/- 0.7, P = 0.001) and those with only albuminuria (8.9 +/- 1.2, P = 0.044). In all subjects taken together there was a strong association between insulin sensitivity and GFR (r = 0.46, P = 0.012). Patients with reduced GFR displayed no significant difference in insulin clearance (12.2 +/- 1.6 mL kg 1 min-1) compared to controls (13.8 +/- 1.3) but a slightly lower insulin clearance than patients with only albuminuria (16.6 +/- 1.0, P = 0.027). There were no significant differences between patient groups in the adipocyte studies in vitro, i.e. with respect to insulin binding, insulin degradation and the effects of insulin on glucose uptake and lipolysis. This is compatible with humoral factors causing whole-body insulin resistance and in the group with reduced GFR, we found that serum parathyroid hormone, interleukin-6 and tumour necrosis factor-alpha levels were elevated whereas the morning cortisol was decreased. CONCLUSIONS: In type 1 diabetes, the appearance of albuminuria does not seem to alter insulin sensitivity and clearance. A marked insulin resistance but no consistent impairment in insulin clearance seems to accompany progression to a stage with a slight reduction in GFR. These alterations are not accompanied by general defects in insulin target cells. Instead, alterations in the regulation of insulin-antagonistic hormones and cytokines could potentially contribute to the development of insulin resistance in diabetic nephropathy. PMID- 11895458 TI - In vitro determination of active bile acid absorption in small biopsy specimens obtained endoscopically or surgically from the human intestine. AB - BACKGROUND: In the construction of a Kock reservoir for continent urinary diversion, 70 cm of the distal ileum are used. Impaired absorption of bile acids in these patients might cause diarrhoea. Data on the absorption of bile acids in different parts of the human intestine are limited. METHODS: Biopsies were taken during endoscopy from the duodenum, the terminal ileum or the right colon, and during surgery 10, 50, 100 and 150 cm proximally to the ileo-caecal valve using standard endoscopy biopsy forceps. The biopsy specimens were incubated in vitro with radio-labelled taurocholic acid at 37 degrees C for 22 or 45 min The radioactivity was determined using the liquid scintillation technique. RESULTS: A linear increase in the uptake was observed, with increased concentrations of taurocholic acid between 100 and 500 microm in all specimens tested, that represented passive uptake or unspecific binding. The active uptake could be calculated from the intercept of the line representing passive uptake with the ordinate. The active uptake in the terminal ileum was 3-4 times greater than 100 cm proximal to the valve. CONCLUSIONS: The active absorption of bile acids in humans can be determined in small biopsy specimens taken using standard biopsy forceps during endoscopy or surgery. This method is suitable for clinical studies of bile acid absorption. Active uptake of bile acids not only takes place in the very distal part of the ileum but also to a considerable degree 100 cm proximally to the ileo-colonic valve. This should be taken into account when selecting the ileal segment for continent urinary diversion. PMID- 11895459 TI - Decreased trabecular bone biomechanical competence, apparent density, IGF-II and IGFBP-5 content in acromegaly. AB - BACKGROUND: Earlier studies on the effect of excess growth hormone (GH) on trabecular bone have been conflicting. Since insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) in part mediate the effects of GH, the present study aimed to investigate trabecular bone composition of these growth factors in relation to biomechanical properties in acromegaly. MATERIALS AND METHODS: Trabecular bone biomechanical competence (compression test), apparent density (peripheral quantitative computed tomography, pQCT), and bone matrix contents of calcium (HCl hydrolysis) and IGFs (guanidinium-HCl extraction) were measured in iliac crest biopsies from 13 patients with active acromegaly (two women and 11 men, aged 21-61 years) and 21 age- and sex-matched controls (four women and 17 men, aged 23-64 years). RESULTS: Trabecular bone pQCT was reduced in acromegalic patients compared with controls (P = 0.005), as was biomechanical competence (P < 0.05 for all measures). These parameters were significantly positively correlated in both acromegalic patients and controls. The calcium content of trabecular bone was significantly increased in patients compared with controls. No significant differences were found in trabecular bone content of IGF I, IGFBP-3, or osteocalcin. However, IGF-II and IGFBP-5 content was decreased (P < 0.001 and P < 0.05, respectively). CONCLUSIONS: The present study demonstrates reduced trabecular biomechanical competence and apparent density in acromegaly, supporting previous observations of an unfavourable effect of chronic excess GH on the axial skeleton. Furthermore, we demonstrate decreased trabecular bone content of IGF-II and IGFBP-5 in these patients. However, we found no direct causal relationship between trabecular bone density and bone content of IGF system components. PMID- 11895460 TI - Antitumour and toxic effects on Wistar rats of two new platinum complexes. AB - BACKGROUND: Cancer chemotherapy in humans based on metal complexes started at the clinical level in the late 1970s with the use of cisplatin, which forms intra strand cross-links with DNA. METHODS: Two new platinum complexes of cis-geometry with the amino acids inosine (ino) and l-alanine (ala), Pt(ino)2Cl2 and cis [Pt(NH3)2(ala)](NO3), respectively, were synthesized and pure samples were obtained by means of flash chromatography. These complexes were tested on benzo(a)pyrene-induced tumours in Wistar rats to detect their antitumour and toxic effects. RESULTS: There was a statistically significant prolongation of the mean survival time of the animals in the two groups tested (272 +/- 18 days and 246 +/- 26 days, respectively) compared to the control group (195 +/- 22 days) (P < 0.001). Toxic effects included a decrease in leucocyte cell count, mild haemolysis, mild haematuria, mild hepatotoxicity, elevated body temperature and hair loss. All of these effects were reversible after drug discontinuation. CONCLUSIONS: The two new platinum complexes described here appear to have an effective antitumour activity without severe toxicity when tested on Wistar rats. PMID- 11895461 TI - A new insight into aspirin-induced asthma. AB - In about 10% of adult patients with asthma, aspirin and other nonsteroidal anti inflammatory drugs precipitate attacks of dyspnoea. Accumulated evidence shows that these reactions are due to the interference of aspirin-like drugs with arachidonic acid metabolism in the lungs of the sensitive patients: inhibition of cyclooxygenase is accompanied by overproduction of cysteinyl leukotrienes. The mechanisms of these reactions and the characteristic course of aspirin-induced asthma and its management are discussed. PMID- 11895462 TI - Absence of p53 gene mutations in skin fibroblasts derived from patients with systemic sclerosis. PMID- 11895463 TI - Oestrogen attenuates coronary vasoconstriction after angioplasty: role of endothelin-1. AB - BACKGROUND AND AIMS: There were controversies as to whether endothelin-1 is released after coronary angioplasty. We sought to determine whether endothelin-1 is released after coronary angioplasty and whether oestrogen administration can affect coronary vasomotor tone by reducing endothelin-1 concentrations. METHODS: The study was designed to prospectively investigate 24 consecutive patients scheduled for elective coronary angioplasty. Patients were randomized into two groups according to whether they did not (group 1, n = 12) or did (group 2, n = 12) have intracoronary treatment with oestrogen. Quantitative coronary angiography was monitored at baseline, immediately after successful angioplasty, and 15 min after the last deflation. Blood samples for measuring the levels of endothelin-1 were drawn from the ascending aorta and the coronary sinus simultaneously before angioplasty and 15 min after balloon dilatation. RESULTS: The diameters of the coronary artery at the dilated segments were significantly reduced 15 min after dilation compared with those immediately after dilation in group 1 from 3.20 +/- 0.22 to 2.30 +/- 0.23 mm (P < 0.001), respectively. The vasoconstriction was significantly blunted in group 2. The endothelin-1 levels from the coronary sinus rose significantly, by 29%, 15 min after angioplasty in group 1, which was attenuated after administering oestrogen. Significant correlation was found between the changes of coronary vasomotion of the dilated segment and endothelin-1 levels (r = 0.70, P = 0.01). CONCLUSION: Endothelin-1 is released into the coronary circulation after angioplasty, and this vasoactive substance may contribute to the occurrence of vasoconstriction. The vasoconstriction is attenuated by oestrogen by reducing the endothelin-1 levels. This finding provided a new strategy to treat coronary vasoconstriction after angioplasty. PMID- 11895465 TI - The effect of the Interleukin-6-174G > C promoter gene polymorphism on endothelial function in healthy volunteers. AB - AIMS: Atherosclerosis is a chronic inflammatory condition, manifest in its early stages by endothelial dysfunction. Interleukin-6 (IL6) plays a key role in driving this process through stimulation of acute phase protein synthesis. We have examined the effect of the IL6 gene -174G > C promoter polymorphism on endothelial function in a group of healthy subjects. METHODS: 248 adults aged 20 28 years participated. Polymerase chain reaction was performed for the -174G > C polymorphism. Brachial artery diameter was measured at rest and after forearm cuff occlusion by high-resolution ultrasound. Responses were represented as absolute flow mediated dilatation (FMDA). RESULTS: Overall there was a trend towards greater FMDA for genotype CC, P = 0.14. No effect was seen in women; however, in men, following multivariate analysis, there was a significant association between genotype and FMDA, P = 0.04. In addition, a significant detrimental effect of smoking on FMDA was only seen in males of genotype CC (P < 0.05) when compared to nonsmokers of the same genotype. CONCLUSION: IL6-174G > C promoter polymorphism influences endothelial function in healthy male subjects. The detrimental effect of smoking on endothelial function is most clearly seen in men of genotype -174 CC, suggesting a genotype-specific interaction with smoking. PMID- 11895464 TI - Elective coronary angioplasty with 60 s balloon inflation does not cause peroxidative injury. AB - BACKGROUND: The aim of this study was to evaluate the ongoing controversial issue of whether ischemia/reperfusion during elective coronary angioplasty evokes myocardial peroxidative injury. DESIGN: We measured indicators of free radical damage to lipids (free malondialdehyde) and proteins (sulphydryl groups) in coronary sinus blood in 19 patients with stable angina who were undergoing elective angioplasty for isolated stenosis of the proximal left anterior descending coronary artery. Ischemia induced by 60 s balloon inflations was confirmed by lactate washout into coronary sinus after deflation, with immediate and 1 min samples. Peroxidative injury was assessed from washout of (a) malondialdehyde measured directly by high performance liquid chromatography and (b) reduced sulphydryl groups, inverse marker of protein oxidative stress. RESULTS: Mean lactate concentration immediately after each deflation increased by 120-150% of the initial value, confirming ischemia and showing that blood originated largely from the ischemic region. Lack of myocardial production of malondialdehyde was confirmed by (a) no arteriovenous differences in individual basal concentrations (aortic, range 0.33-12.03 nmol mL-1, mean 7.82; coronary sinus blood, range 0.52-15.82 nmol mL-1, mean 8.18), and (b) after deflations, mean concentrations were not significantly different from preocclusion value. There was no decrease in concentration of sulphydryl groups throughout angioplasty. CONCLUSION: Elective coronary angioplasty with 60 s balloon inflations is a safe procedure that does not induce peroxidative myocardial injury as assessed by methods used in the present study. PMID- 11895466 TI - Helicobacter pylori masks differences in homocysteine plasma levels between controls and type 2 diabetic patients. AB - BACKGROUND: Data in the literature have not clarified whether type 2 diabetes mellitus affects homocysteine plasma levels. Different variables able to influence homocysteine could be the cause of these controversial findings. An important but neglected confounding factor is Helicobacter pylori, which has been demonstrated to be a cause of elevated levels of homocysteine and which is prevalent in the Caucasian population, ranging from 30 to 40% incidence. Starting from these findings we wanted to verify whether differences in homocysteine levels exist between a type 2 diabetic population and a control group, taking into account the presence/absence of Helicobacter pylori. DESIGN: The study was carried out on a group of uncomplicated and normotensive type 2 diabetic patients (n = 30, 55.7 +/- 9.7 years) and on a control group (n = 43, 51.2 +/- 11.3 years). On these subjects we evaluated: main parameters of glyco- and lipo metabolic balance, presence of Helicobacter pylori by 13C Urea Breath Test, plasma homocysteine, vitamin B12, folate and genetic polymorphism of methylenetetrahydrofolate reductase. RESULTS: Evaluating the two groups as a whole, significant differences in homocysteine were found when considering Helicobacter pylori presence/absence (14.0 +/- 6.5 vs. 10.6 +/- 4.7 micromol L-1, respectively, P < 0.01) without differences of vitamins and the genetic polymorphism of methylenetetrahydrofolate reductase. The positive interaction found among Helicobacter pylori, diabetes and homocysteine (P = 0.03) taking into account all the other evaluated confounding factors, demonstrates that a significant difference in homocysteine plasma levels exists between diabetics and controls (Helicobacter pylori-negative: diabetics 12.5 +/- 5.6 micromol L-1, controls 9.4 +/- 3.8 micromol L-1; Helicobacter pylori-positive: diabetics 13.6 +/- 5.8 micromol L-1, controls 14.3 +/- 7.0 micromol L-1). CONCLUSIONS: Type 2 diabetes seems to induce per se higher levels of homocysteine, which appears to be one of the factors responsible for the increased risk of vascular damage. PMID- 11895467 TI - Microcirculatory effects of KATP channel blockade by sulphonylurea derivatives in humans. AB - BACKGROUND: Recent investigations have shown that glibenclamide inhibits the opening of vascular ATP-sensitive potassium channels during ischemia. This observation may implicate cardiovascular effects of sulphonylurea derivatives when used under conditions of ischemia in patients with Type 2 diabetes mellitus. In addition to resistance arteries, the (pre) capillary vessels also contain ATP dependent potassium channels. Closure of these channels by sulphonylurea derivatives might affect the development of microvascular disease in Type 2 diabetes mellitus. Therefore, we investigated the microcirculatory effects of sulphonylurea derivatives in Type 2 diabetic patients as compared with healthy volunteers. MATERIALS AND METHODS: Arteriovenous blood flow (skin temperature and laser Doppler flux) and capillary blood cell velocity were measured before and during infusion of four doses of glibenclamide (0.1, 0.3, 1.0 and 3.0 microg min 1 dL-1) into the brachial artery of 14 Type 2 diabetic patients and 13 healthy controls. The experiments included appropriate time control studies. RESULTS: Both skin temperature and laser Doppler flux decreased in response to glibenclamide in healthy volunteers (-7 +/- 2%, P < 0.0005 and -31 +/- 11%, P = 0.001, respectively), but did not change in Type 2 diabetic patients (1 +/- 3%, P = 0.29 and 4 +/- 14%, P = 0.97). However, capillary blood cell velocity decreased in Type 2 diabetic patients (-38 +/- 18%, P = 0.04), but did not change in healthy volunteers (-1 +/- 11%, P = 0.28). CONCLUSIONS: The results of the present study indicate that glibenclamide indeed affects microvascular blood flow. Glibenclamide may induce redistribution of the microvascular skin flow from nutritive flow to arteriovenous shunt flow in Type 2 diabetic patients. Therefore, closure of ATP-dependent potassium channels by glibenclamide possibly plays a role in the development of microangiopathy in Type 2 diabetic patients. PMID- 11895468 TI - Endocrine abnormalities in healthy first-degree relatives of type 2 diabetes patients--potential role of steroid hormones and leptin in the development of insulin resistance. AB - BACKGROUND: First-degree relatives of type 2 diabetes patients are at risk of developing diabetes and they display several metabolic and hormonal perturbations. The interplay between insulin resistance, steroid hormones and circulating leptin is, however, still not fully explored in this group. DESIGN: Thirty-three healthy first-degree relatives of type 2 diabetic patients (relatives; M/F 19/14) were compared to 33 healthy subjects without a family history of diabetes (controls) and the groups were matched for gender, age and body mass index (BMI). We performed euglycaemic hyperinsulinaemic clamps and blood was sampled for hormone analyses. RESULTS: Relatives exhibited decreased insulin sensitivity (index of metabolic clearance rate of glucose; MCRI) but when genders were analysed separately, this difference was significant only in males (11.3 +/- 1.3 vs. 15.0 +/- 1.5 units, means +/- SEM, P = 0.030). In male relatives morning cortisol and testosterone levels were lower, whereas leptin was higher than in male controls (P = 0.018, 0.008 and 0.063, respectively). In male relatives plasma testosterone levels were significantly associated with insulin sensitivity (r = 0.48, P = 0.040). Circulating leptin levels were inversely correlated with insulin sensitivity in all subject groups (r-values -0.49 to 0.66; P < 0.05, except in female control subjects P = 0.063). These associations were present also when age and BMI or waist:hip ratio were included in stepwise multiple regression analyses. CONCLUSION: Male subjects genetically predisposed for type 2 diabetes display several endocrine abnormalities including leptin, cortisol and testosterone levels. Dysregulation of these hormones may be important in the development of insulin resistance and type 2 diabetes. PMID- 11895469 TI - Blood glucose pre-prandial baseline decreases from morning to evening in type 2 diabetes: role of fasting blood glucose and influence on post-prandial excursions. AB - BACKGROUND: To know the relationships between pre- and postprandial blood glucose (BG), i.e. BG profile shape, is a requisite for an appropriate therapy for type 2 diabetic patients. In non diabetic subjects, pre-breakfast, pre-lunch and pre dinner BG are similar, so that BG postprandial excursions are superimposed on a stable BG preprandial baseline. We aimed to clarify: (a) whether BG preprandial baseline is stable also in type 2 diabetes and (b) whether fasting BG (FBG) influences the slope of BG preprandial baseline and the relationships between pre and postprandial BG. DESIGN: We evaluated self-measured BG profiles of 237 type 2 diabetic patients on diet alone (M/F, 152/85; age 58.6 +/- 0.7 years; years from diagnosis 4.8 +/- 0.6; BMI 28.0 +/- 0.3 kg m-2): 536 profiles containing preprandial BG (corresponding HbA1c 6.8 +/- 0.06%) and 208 profiles containing both pre- and postprandial BG (corresponding HbA1c 6.8 +/- 0.09%). The profiles, measured by nurses, of 866 type 2 diabetic patients on diet alone were also considered (corresponding HbA1c 6.7 +/- 0.04%). RESULTS: In self-measured profiles containing only preprandial BG: (i) FBG (6.77 +/- 0.07 mmol L(-1)) is higher than pre-lunch BG (6.09 +/- 0.07 mmol L(-1)), P = 0.0001) and pre-dinner BG (5.84 +/- 0.06 mmol L(-1)), P =0.0001); (ii) the delta value between FBG and pre-dinner BG is correlated with FBG (r = 0.57, P = 0.0001), the highest FBG, the steepest the fall of BG preprandial baseline throughout the day. This trend is confirmed in profiles measured by nurses. In profiles containing both pre- and postprandial BG: (i) there is a trend to preprandial BG fall (P = 0.0001) and to postprandial BG increase (P = 0.0001) from morning to evening; (ii) postprandial excursions are influenced and sometimes masked by the slope of BG preprandial baseline, thus, in profiles with FBG < or = 6.7 mmol L(-1), all postprandial values are higher than FBG (P = 0.0001), whereas in profiles with FBG > 7.8 mmol L(-1), postprandial values are not significantly higher than FBG. CONCLUSION: In type 2 diabetes, the shape of BG profiles changes in relation to FBG, because it deeply influences the slope of BG preprandial baseline on which postprandial excursions are superimposed. Thus, before planning treatment policies, not only the extent of fasting and postprandial hyperglycaemia, but also the shape of profiles should be considered, to safely correct hyperglycaemia without inducing hypoglycaemia. PMID- 11895470 TI - Alcohol consumption alters insulin secretion and cardiac autonomic activity. AB - BACKGROUND: Alcohol may have a cardioprotective effect. One possible mechanism is by modifying insulin resistance/secretion. The aims of this study were: (i) to examine the effect of short-term alcohol consumption on the metabolic control of glucose tolerance; (ii) to study the influence of short-term alcohol consumption on cardiac autonomic activity using spectral analysis of heart rate variability. METHODS: Twenty-one healthy subjects, in a randomized crossover design, either received three units of ethanol daily for 1 week or abstained from ethanol. The control of glucose tolerance was assessed using the intravenous glucose tolerance test with minimal modelling. RESULTS: There was no difference in fasting glucose, fasting insulin or insulin sensitivity between the two groups. Alcohol showed a lower insulin first phase insulin response (no alcohol 659.0 +/- 394.1 SD, alcohol 535.2 +/- 309.1) pmol L-1 min-1, P = 0.027). There was no difference in heart rate or blood pressure but a significant difference in the ratio of high to low frequency spectral power of heart rate variability; (no alcohol 4.55 +/- 3.78, alcohol 8.16 +/- 6.77, P = 0.033). This suggests decreased sympathetic and/or increased vagal modulation of heart rate in the alcohol group. CONCLUSION: The finding of no difference in insulin sensitivity between the two groups contrasts with, but does not entirely contradict, the results of previous epidemiological studies--perhaps suggesting that longer term changes such as liver enzyme induction may be important. The difference in insulin secretion questions the validity of previous studies of the influence of alcohol on insulin sensitivity, where insulin levels were used as a surrogate for insulin resistance. PMID- 11895471 TI - Increased TGFbeta1 plasma level in patients with lung cancer: potential mechanisms. AB - BACKGROUND: Plasma transforming growth factor beta1 (TGFbeta1) levels are elevated in patients with lung cancer. As TGFbeta1 is mainly found in platelets and as nonmalignant pulmonary diseases (NMPD) are frequently associated with lung cancer, we investigated the potential contribution of platelet degranulation and/or of a concomitant NMPD to the increased plasma levels of TGFbeta1 reported in patients with lung cancer. MATERIALS AND METHODS: Blood samples were collected in duplicate from 30 healthy subjects, 14 patients suffering from NMPD and 37 patients with lung cancer. The platelet count was determined and the samples were processed to obtain plasma. One sample was collected in EDTA (EDTA plasma) and the other in a mixture inhibiting platelet degranulation (PIM plasma). TGFbeta1 concentrations and beta-thromboglobulin (betaTG) levels, an index of platelet degranulation, were measured in both plasma samples. RESULTS: TGFbeta1 and betaTG plasma levels measured in PIM plasma were lower than those obtained in EDTA plasma. With respect to PIM plasma, both TGFbeta1 and betaTG levels were higher in patients with lung cancer than those with NMPD and in healthy individuals. In patients with NMPD, only TGFbeta1 levels were increased as compared to healthy controls, betaTG levels being similar. CONCLUSION: Methods for collecting and processing blood samples are critical in determining reliable circulating TGFbeta1 levels. Increased TGFbeta1 plasma levels observed in patients with lung cancer are related, at least partly, to concomitant NMPD and also to platelet degranulation as proved by increased betaTG levels. PMID- 11895472 TI - Angiogenesis extent and expression of matrix metalloproteinase-2 and -9 agree with progression of ovarian endometriomas. AB - BACKGROUND: Changes in angiogenesis and expression of extracellular matrix degrading enzymes have been substantiated in tumour changeover and progression. METHODS: Tissues from 44 biopsies of stage III and IV ovarian endometriomas, and 10 biopsies of normal (control) endometrium were investigated immunohistochemically to count microvessels, and by in situ hybridization to assess the expression of mRNA of matrix metalloproteinase-2 (MMP-2) and MMP-9. Implants of the tissues were investigated in the chick embryo chorioallantoic membrane (CAM) assay to determine their angiogenic capacity. RESULTS: The endometriomas displayed significantly higher counts than normal endometria and the highest values were associated with the deepest invasion level (stage IV). Microvessels localized in close association with ectopic endometrial cells in the form of winding and arborized tubes, often dilated in microaneurysmatic segments. These were absent in normal endometrium. Expression of MMP-2 and MMP-9 mRNA, evaluated as percentages of positive biopsies and intensity of expression, was up regulated in endometriomas and more pronounced in stage IV. MMP-2 and MMP-9 mRNA were also expressed by host stromal cells, including microvascular endothelial cells, fibroblasts and macrophages, whereas the control endometrium showed very little expression of MMP-2 mRNA in a few endothelial cells and no expression of MMP-9 mRNA. Implants from stage IV endometrioma induced a more intense vasoproliferative response than those from stage III, while no vasoproliferative response was induced by the normal endometrium. CONCLUSION: These data suggest that angiogenesis and degradation of extracellular matrix occur together in endometriosis and are more pronounced in stage IV, and that endometriosis cells and some host stromal cell populations co-operate in disease progression. PMID- 11895473 TI - Tumour cell-dendritic cell fusion for cancer immunotherapy: comparison of therapeutic efficiency of polyethylen-glycol versus electro-fusion protocols. AB - BACKGROUND: Fusion of tumour cells with dendritic cells (DC) is a powerful new technology to increase tumour vaccine immunogenicity. The aim of this study was to compare fusion protocols with syngenic DCs with respect to the efficiency of polyethylen-glycol-(PEG) and electric pulse-mediated fusions for induction of protective anti-tumour immune responses. As a model we chose a low immunogenic and metastatic murine mammary carcinoma cell line, which mimics clinically relevant tumour features. METHODS: FACS-staining, chromium release assay, therapeutic immunization, adoptive transfer. RESULTS: We show that the parental line with low cell surface expression of MHC molecules as well as a lacZ transfectant becomes highly immunogenic upon fusion with DCs. This was true for PEG- as well as for electro-fused cells. Immunization with products of DCs and tumour cells cocultivated for 16 h without the fusing agent PEG also caused induction of profound anti-tumour immunity, while this was not the case when using parental tumour cells or their lacZ transfectants as vaccines. Immune protection against the parental tumour cells after vaccination with fused cells was long-lasting and could be transferred via immune spleen cells into immuno incompetent nude (nu/nu) mice. CONCLUSION: Fusion products of DA3(hi) mammary carcinoma cells and DCs produced by an electric pulse were similar to those produced by PEG fusion with regard to vaccine potency in prophylactic antitumour immunization assays in vivo. Therefore, both techniques seem to be promising for clinical application. PMID- 11895474 TI - Phosphorylation and reorganization of vimentin by p21-activated kinase (PAK). AB - BACKGROUND: Intermediate filament (IF) is one of the three major cytoskeletal filaments. Vimentin is the most widely expressed IF protein component. The Rho family of small GTPases, such as Cdc42, Rac and Rho, are thought to control the organization of actin filaments as well as other cytoskeletal filaments. RESULTS: We determined if the vimentin filaments can be regulated by p21-activated kinase (PAK), one of targets downstream of Cdc42 or Rac. In vitro analyses revealed that vimentin served as an excellent substrate for PAK. This phosphorylated vimentin lost the potential to form 10 nm filaments. We identified Ser25, Ser38, Ser50, Ser65 and Ser72 in the amino-terminal head domain as the major phosphorylation sites on vimentin for PAK. The ectopic expression of constitutively active PAK in COS-7 cells induced vimentin phosphorylation. Fibre bundles or granulates of vimentin were frequent in these transfected cells. However, the kinase-inactive mutant induced neither vimentin phosphorylation nor filament reorganization. CONCLUSION: Our observations suggest that PAK may regulate the reorganization of vimentin filaments through direct vimentin phosphorylation. PMID- 11895475 TI - Replication fork block protein, Fob1, acts as an rDNA region specific recombinator in S. cerevisiae. AB - BACKGROUND: The analysis of homologous recombination in the tandemly repeating rDNA array of Saccharomyces cerevisiae should provide useful information about the stability of not only the rDNA repeat but also the abundant repeated sequences on higher eukaryotic genomes. However, the data obtained so far are not yet conclusive, due to the absence of a reliable assay for detecting products of recombination in the rDNA array. RESULTS: We developed an assay method to detect the products of unequal sister-chromatid recombination (marker-duplication products) in yeast rDNA. This assay, together with the circular rDNA detection assay, was used for the analysis. Marker-duplication occurred throughout the rDNA cluster, preferentially between nearby repeat units. The FOB1 and RAD52 genes were required for both types of recombinant formation. FOB1 showed a gene dosage effect on not only the amounts of both recombinants, but also on the copy number of the repeat. However, unlike the RAD52 gene, the FOB1 gene was not involved in homologous recombination in a non-rDNA locus. In addition, the marker-duplication products were drastically decreased in the mre11 mutant. CONCLUSION: Our data demonstrate that FOB1- and RAD52-dependent homologous recombination cause the gain and loss of a few copies of the rDNA unit, and this must be a basic mechanism responsible for amplification and reduction of the rDNA copy number. In addition, FOB1 may also play a role in the copy number regulation of rDNA tandem repeats. PMID- 11895476 TI - Isolation and characterization of a new nucleolar protein, Nrap, that is conserved from yeast to humans. AB - BACKGROUND: The nucleolus is the site of rRNA synthesis and processing in eukaryotic cells, but its composition remains poorly understood. RESULTS: We have identified a novel nucleolar RNA-associated protein (Nrap) which is highly conserved from yeast (Saccharomyces cerevisiae) to human, with homologues in mouse, Drosophila melanogaster, Caenorhabditis elegans, Arabidopsis thaliana, Schizosaccharomyces pombe, and other species. In the mouse, we have found that Nrap is ubiquitously expressed and is specifically localized in the nucleolus. We have also identified splice variants in human and mouse, and defined the intron exon structure of the human Nrap gene. Nrap is inherited into daughter nuclei by associating with the condensed chromosomes during mitosis. RNase treatment of permeabilized cells indicated that the nucleolar localization of Nrap is RNA dependent. The effects of actinomycin D, cycloheximide and 5,6-dichloro-beta-d ribofuranosyl-benzimidazole on Nrap expression and distribution in cultured cells suggest that Nrap is associated with the pre-rRNA transcript. CONCLUSIONS: Nrap is a large nucleolar protein (of more than 1000 amino acids), and is a new class of protein with new structural and functional motifs. Nrap appears to be associated with ribosome biogenesis by interacting with pre-rRNA primary transcript. PMID- 11895477 TI - Regulation of alternative splicing of alpha-actinin transcript by Bruno-like proteins. AB - BACKGROUND: The Bruno-like or CELF proteins, such as mammalian CUGBP1 and Etr-3, Xenopus EDEN-BP, and Drosophila Bruno (Bru), are regulators of gene expression at the post-transcriptional level, and contain three RNA-recognition motifs (RRMs). It has been shown that mammalian CUGBP1 and Etr-3 regulate alternative splicing of cardiac troponin T pre-mRNA via binding to CUG-triplet repeats. RESULTS: Using in vitro selection and UV-crosslinking experiments, we found that zebrafish Bruno like proteins bound to repeat elements of uridine and purine (termed UREs). It is known that non-muscle (NM) and smooth muscle (SM) exons of the rat alpha-actinin gene are used in a mutually exclusive manner. Transfection experiments in mammalian cells showed that zebrafish Brul and Etr-3 induced the muscle-specific splicing of rat alpha-actinin pre-mRNA via binding to the URE at the branch point upstream of the NM exon. In contrast, zebrafish Etr-1 promoted skipping of both the NM and SM exons in a manner which was not dependent on URE-binding. CONCLUSIONS: Our results showed that Bruno-like proteins bind to UREs and regulate the alternative splicing of alpha-actinin pre-mRNA. Members of the Bruno family play multiple roles in splicing regulation. PMID- 11895478 TI - Hypoxia-inducible factor-1alpha induces cell cycle arrest of endothelial cells. AB - BACKGROUND: Hypoxia can induce tissue injury, including apoptosis of endothelial cells. However, little is known about the effects of hypoxia on endothelial cell function. We assessed the effects of hypoxia inducible factor (HIF)-1alpha on the functional characteristics of endothelial cells, particularly on cell cycle regulators, by cationic liposome-mediated transfection of HIF-1alpha-expression vector into the cells. RESULTS: Transfection of the HIF-1alpha gene in endothelial cells resulted in (a) reduced proliferation and detachment of the cells; (b) up-regulation of intracellular p21waf1/cip1 and down-regulation of bcl 2; (c) reduced activities of cyclin-dependent kinase (CDK)-4 and CDK-6; (d) cell cycle arrest at G0/G1 phase; and (e) apoptosis of the cells. CONCLUSIONS: HIF 1alpha can induce cell cycle arrest, resulting in the reduced proliferation and apoptosis of endothelial cells, and the hypoxia-induced cell death may be involved by suppression of anti-apoptotic molecule, bcl-2. PMID- 11895479 TI - Hyperploidy induced by drugs that inhibit formation of microtubule promotes chromosome instability. AB - BACKGROUND: Antimicrotubule drugs (AMDs), such as taxol and vincristine, are the most important addition to the chemotherapeutic armamentarium against human cancers. It has been shown that prolonged AMD treatment induces hyperploidy in G1 checkpoint-defective cancer cells and that these hyperploid cells subsequently undergo apoptosis. However, a fraction of these hyperploid cells are able to survive the prolonged mitotic stress and resume cell-cycle progression. RESULTS: We established hyperploid clones that escaped from cell death after AMD treatment from two glioma cell lines, U251MG and U87MG. Subtractive comparative genomic hybridization (CGH) analysis revealed that clones derived from U87MG mainly had chromosome number changes, but that those from U251MG showed both numerical and structural chromosomal changes. Furthermore, numerous aberrations identified in U251MG clones were remarkably chromosome-specific, which may have been due to clonal selection for cells that have an advantage in growth and/or survival. All clones derived from both cell lines had abnormalities in chromosome segregation, and karyotypes of clones were more heterogeneous than those of parental cells, suggesting that cells having a higher chromosome number are subject to asymmetric chromosome segregation, resulting in a heterogeneous karyotype. All clones derived from U87MG and U251MG increased both centric and acentromeric micronuclei, suggesting the presence of chromosome structural abnormality. CONCLUSIONS: AMD treatment induces hyperploid formation and chromosome instability in checkpoint-deficient cancer cells. PMID- 11895480 TI - Restoration of circadian behavioural rhythms in a period null Drosophila mutant (per01) by mammalian period homologues mPer1 and mPer2. AB - BACKGROUND: Recent molecular studies suggest that mammals and Drosophila utilize similar components to generate circadian (approximately equal to 24 h) rhythms. The first identified circadian clock gene, the period (per) gene, is indispensable for behavioural rhythms in Drosophila and is represented in mammals by three orthologues, the relative roles of which are not known. In this study, we investigated the functional conservation of per by introducing the mouse mPer1 and mPer2 genes, driven by the Drosophila timeless (tim) promoter, into Drosophila melanogaster. RESULTS: Behavioural assays showed that both mPer constructs restored rhythms in per(01) flies that are otherwise arrhythmic due to a lack of endogenous per protein (PER). However, the rhythms restored by mPer2 were generally stronger and differed in periodicity from those restored by mPer1. In rhythmic transgenic flies, mPER proteins were expressed in lateral neurones and/or many cells in optic lobe. In addition, cell culture experiments indicated that the Drosophila PER partner, TIM, can form a complex with each of these two mammalian proteins. CONCLUSIONS: This study demonstrates that both mPer1 and mPer2 can function as clock components, and has implications for a functional analysis of the different per genes. PMID- 11895481 TI - Identification of a novel nonlysosomal sulphatase expressed in the floor plate, choroid plexus and cartilage. AB - BACKGROUND: Sulphated glycosaminoglycans (GAGs) attached to proteoglycan core proteins are implicated in cell adhesion, motility and morphogenesis. Variable sulphation patterns, which are thought to be important for regulating proteoglycan function, are generated by sequential reactions during GAG biosynthesis. However, the mechanism by which such diversity is generated remains unclear. RESULTS: A novel sulphatase, designated RsulfFP1, was isolated from rat embryos by screening for floor plate specific genes. RsulfFP1 and its orthologues show homology with other sulphatases, and have a distinctive hydrophilic insertion. In situ hybridization showed that RsulfFP1 mRNA is strongly expressed in the floor plate, choroid plexus and cartilage in rat embryos. In vitro transfection experiments revealed that the RsulfFP1 protein is localized to the Golgi apparatus and endoplasmic reticulum, and is not present in the lysosomes. It also appears to be localized on the cell surface. CONCLUSIONS: RsulfFP1, a phylogenetically conserved sulphatase, forms a novel subgroup in the sulphatase family. It shows homology with the lysosomal sulphatases involved in GAG degradation. Localization of the RsulfFP1 protein in the Golgi apparatus and on the cell surface, however, suggests that it may play a role in regulating proteoglycan-mediated signalling by the desulphation of GAGs during biosynthesis or after GAGs are presented in the extracellular space. PMID- 11895482 TI - Identification of activity-regulated proteins in the postsynaptic density fraction. AB - BACKGROUND: The postsynaptic density (PSD) at synapses is a specialized submembranous structure where neurotransmitter receptors are linked to cytoskeleton and signalling molecules. Activity-dependent dynamic change in the components of the PSD is a mechanism of synaptic plasticity. Identification of the PSD proteins and examination of their modulations dependent on synaptic activity will be valuable for an understanding of the molecular basis of learning and memory. RESULT: We attempted here to identify proteins in the PSD fraction by two-dimensional (2D) gel electrophoresis and mass spectrometry. About 1.7 x 103 protein spots were detected on 2D gels. A total of 90 spots were identified, containing 47 different protein species. In addition to previously identified PSD proteins such as PSD-95/SAP90, several new proteins were identified in the PSD fraction. They included stomatin-like protein 2 and NIPSNAP1. We also examined activity-dependent modulations of PSD proteins by 2D gel electrophoresis. The spot concentration of G protein beta subunit 5 and NIPSNAP1 increased 2 h after kainate treatment that caused generalized seizures. CONCLUSION: These results indicate that the combination of 2D gel electrophoresis and mass spectrometry is an excellent tool for the identification of activity-regulated PSD proteins. PMID- 11895483 TI - Phosphatidylinositol 3-phosphate 5-kinase is required for the cellular response to nutritional starvation and mating pheromone signals in Schizosaccharomyces pombe. AB - BACKGROUND: Phosphatidylinositol (3,5) bisphosphate, which is converted from phosphatidylinositol 3-phosphate by phosphatidylinositol 3-phosphate 5-kinase, is implicated in vacuolar functions and the sorting of cell surface proteins within endosomes in the endocytic pathway of budding yeast. A homologous protein, SpFab1p, has been found in the fission yeast Schizosaccharomyces pombe, but its role is not known. RESULTS: Here we report that SpFab1p is encoded by ste12+ known as a fertility gene in S. pombe. The ste12 mutant grew normally under stress-free conditions, but was highly vacuolated and swelled at high temperatures and under starvation conditions. In nitrogen-free medium, ste12 cells were arrested in G1 phase, but partially defective in the expression of genes responsible for mating and meiosis. The ste12 mutant was defective both in the production of, and in the response to, mating pheromones. The amount of the pheromone receptor protein Map3p, was substantially decreased in ste12 cells. Map3p was transported to the cell surface, then internalized and eventually transported to the vacuolar lumen, even in the ste12 mutant. CONCLUSION: The results indicate that phosphatidylinositol(3,5)bisphosphate is essential for cellular responses to various stresses and for the mating pheromone signalling under starvation conditions. PMID- 11895484 TI - Meu10 is required for spore wall maturation in Schizosaccharomyces pombe. AB - BACKGROUND: Many genes are meiosis and/or sporulation-specifically transcribed during this process. Isolation and analysis of these genes might help us to understand how meiosis and sporulation are regulated. For this purpose, we have isolated a large number of cDNA clones from Schizosaccharomyces pombe whose expression is up-regulated during meiosis. RESULTS: We have isolated meu10+ gene, which encodes 416 amino acids and bears homology to SPS2 of Saccharomyces cerevisiae. A strain whose meu10+ gene has been deleted forms no viable spores. Thin-section electron micrographs showed that the meu10Delta strain has abnormally formed spore walls, and then they disrupt, allowing cytoplasmic material to escape. The Meu10-GFP fusion protein is localized to the spore periphery, thereafter returned to the cytoplasm after sporulation. Meu10-GFP localization to the spore wall was almost normal in the bgs2Delta or chs1Delta mutants that lack 1,3-beta-glucan or chitin, respectively. In contrast, 1,3-beta glucan is abnormally localized in meu10Delta cells. Meu10 has an N-terminal domain with homology to the mammalian insulin receptor and a C-terminal domain with a transmembrane motif. Mutants whose N-terminal or C-terminal domain was truncated were severely defective for sporulation. CONCLUSIONS: Meu10 is a spore wall component and plays a pivotal role in the formation of the mature spore wall structure. PMID- 11895485 TI - Is detection of endocrine cells in breast adenocarcinoma of diagnostic and clinical significance? PMID- 11895486 TI - Neuroendocrine differentiation and prognosis in breast adenocarcinoma. AB - AIMS: Neuroendocrine differentiation has been detected, and its prognostic value studied, in a number of common human carcinomas. To date there are few detailed studies examining its relevance in breast carcinoma. In this study we evaluate the frequency and prognostic importance of neuroendocrine differentiation in breast adenocarcinoma. METHODS AND RESULTS: The presence of neuroendocrine differentiation, defined as positive reactivity for three markers, neuron specific enolase (NSE), chromogranin A and/or synaptophysin, has been evaluated in 99 patients with primary operable breast cancer using standard immunocytochemical techniques. A consecutive cohort of patients were selected from the Nottingham/Tenovus series. Comprehensive patient and tumour records have been maintained, and patients were followed up according to a defined protocol. Eighteen cases were positive for NSE, 10 for chromogranin A and 13 for synaptophysin. Eleven percent were positive with more than one neuroendocrine marker. No significant association was found between neuroendocrine differentiation and tumour size, grade, stage or the prevalence of vascular invasion. There was no significant difference in either overall or disease-free survival between patients with or without neuroendocrine differentiation. CONCLUSIONS: In this study we confirm that neuroendocrine differentiation can be identified in a subset (10-18%) of human breast carcinomas. This phenomenon appears to have no relationship to established prognostic factors or patient outcome. PMID- 11895487 TI - Secretory carcinoma of the breast: a tumour analogous to salivary gland acinic cell carcinoma? AB - AIMS: Acinic cell-like breast carcinoma is a newly recognized entity, and few acinic cell-like breast carcinoma cases have been reported. All reported acinic cell-like breast carcinomas were counterparts of the solid type of acinic cell carcinoma of the salivary gland. We report here three cases of secretory breast carcinoma with acinic cell differentiation, and discuss the similarity between secretory breast carcinoma and acinic cell carcinoma of the salivary gland. METHODS AND RESULTS: The cases were histologically identical to acinic cell carcinoma of the salivary gland: papillary-cystic type in case 1, a mixture of papillary-cystic, microcystic and follicular type in case 2, and microfollicular type in case 3. Immunohistochemically, the tumour cells were positive for salivary-type amylase, lysozyme, S100 protein and alpha 1-antitrypsin, and negative or less reactive for gross cystic disease fluid protein-15 and oestrogen receptor. All three cases did not reveal metastasis or recurrence. CONCLUSIONS: These cases were typical of secretory breast carcinoma, and were clinically, histologically and immunohistochemically analogous to acinic cell carcinoma of the salivary gland. We emphasize that secretory breast carcinoma and acinic cell carcinoma of the salivary gland may be identical lesions. PMID- 11895488 TI - Intraepidermal cytokeratin 7 immunoreactive cells in the non-neoplastic nipple may represent interepithelial extension of lactiferous duct cells. AB - AIMS: The interpretation of cytokeratin 7 (CK7)-positive cells in the epidermis of the nipple has been controversial. These cells have been described in Paget's disease of the nipple, and they have also been cited as benign 'Toker' cells or as Merkel cells. Having observed CK7+ cells in histologically unremarkable nipple biopsies, we sought to assess the distribution of CK7+ cells in Paget's disease of the nipple and in histologically unremarkable nipple. METHODS AND RESULTS: Representative sections from 37 cases of Paget's disease of the nipple and 32 cases of histologically unremarkable nipple were obtained. The histologically unremarkable nipple sections were taken from prophylactic mastectomies (n=17) and from autopsies of patients who did not have breast cancer (n=15). CK7 immunostaining was performed on sections from formalin-fixed paraffin blocks. Sequential sections were immunostained with antibodies to low-molecular weight cytokeratin-CAM 5.2 and HER-2/neu. CK7+ cells were present in the epidermis around the opening of the lactiferous ducts in Paget's disease (95%) and in histologically unremarkable nipple (45%) cases. CK7+ cells diminished in number with increasing distance from the orifice of the lactiferous ducts. The lactiferous duct epithelium in Paget's disease and in histologically unremarkable nipple was CK7+ in all specimens when this element was present. CAM5.2 immunostaining had a similar but weaker pattern of reactivity. HER-2/neu reactivity was seen in 68% cases of Paget's disease and was negative in all cases of histologically unremarkable nipple. Tumour cells in two cases of Paget's disease were CK7-. In one of these, the underlying breast carcinoma was also CK7 , the only CK7- tumour in this series. In the other case, the normal lactiferous duct was CK7+ and no underlying carcinomatous tissue was available to study. CONCLUSIONS: The presence of CK7+ cells does not equate to Paget's disease of the nipple. Intraepidermal CK7+ cells in the non-neoplastic nipple can be a manifestation of interepithelial extension of benign lactiferous duct cells. The increased presence of CK7+ cells in Paget's disease probably results either from neoplastic transformation of native intraepithelial lactiferous duct cells or form direct extension/migration of neoplastic cells into the nipple. The distribution of CK7 immunoreactive cells in the nipple epidermis can be helpful in the diagnosis of Paget's disease of the nipple. PMID- 11895490 TI - Cotyledonoid hydropic intravenous leiomyomatosis: a new variant leiomyoma. AB - AIMS: We present the histopathological findings of a series of six cases of a benign uterine smooth muscle tumour with an unusual growth pattern. METHODS AND RESULTS: All cases have the appearances of the recently described dissecting (cotyledonoid) leiomyoma. In addition, three of these lesions demonstrate the features of intravenous leiomyomatosis with varying degrees of hydropic degeneration. CONCLUSIONS: This combination of phenotypes has not previously been described within the literature; therefore we propose that these are classified as examples of 'cotyledonoid hydropic intravenous leiomyomatosis', a new variant of unconventional leiomyoma. PMID- 11895489 TI - Value of mesothelial and epithelial antibodies in distinguishing diffuse peritoneal mesothelioma in females from serous papillary carcinoma of the ovary and peritoneum. AB - AIMS: To evaluate the role of mesothelial markers (calretinin, thrombomodulin, cytokeratin 5/6, and CD44H) and carcinoma markers (polyclonal and monoclonal carcinoembryonic antigen, Leu-M1, CA-125 and Ber-EP4) in distinguishing diffuse peritoneal malignant mesothelioma from primary serous papillary adenocarcinoma of the ovary and peritoneum. METHODS AND RESULTS: Paraffin-embedded formalin-fixed blocks from 32 diffuse peritoneal mesotheliomas of epithelial subtype (all females), 20 serous papillary ovarian carcinomas and three primary peritoneal serous papillary carcinomas were studied. Calretinin and Ber-EP4 appeared to be the best positive mesothelial and carcinoma marker, respectively. Nuclear calretinin expression was identified in 28 of 32 malignant mesotheliomas with no nuclear immunoreactivity in the cohorts of serous papillary ovarian and peritoneal carcinomas, thus yielding 88% sensitivity and 100% specificity. Ber EP4 showed 95% sensitivity and 91% specificity for serous papillary ovarian carcinoma. Thrombomodulin, cytokeratin 5/6 and CD44H immunoreactivities were seen in 18 (56%), 17 (53%) and 15 (47%) of peritoneal mesotheliomas, respectively, and in six (30%), five (25%) and five (25%) of the ovarian tumours, respectively. None of the three primary peritoneal serous papillary carcinomas expressed calretinin, thrombomodulin, cytokeratin 5/6 or CD44H. Polyclonal and monoclonal CEA, and Leu-M1 were expressed by two (10%), one (5%) and seven (35%) serous papillary ovarian carcinomas, respectively. None of the serous papillary peritoneal carcinomas expressed polyclonal CEA, monoclonal CEA or Leu-M1. CA-125 was positive in 19 (95%) and two (67%) ovarian and peritoneal carcinomas, respectively, and in eight (25%) peritoneal mesotheliomas. CONCLUSIONS: Calretinin and Ber-EP4 are useful discriminant markers in distinguishing peritoneal mesothelioma in women from serous papillary ovarian and peritoneal carcinoma. The other mesothelial markers (thrombomodulin, cytokeratin 5/6, and CD44H) and carcinoma markers (polyclonal and monoclonal CEA, and Leu-M1) yielded a too low sensitivity for practical use. PMID- 11895491 TI - Expression of angiogenic factors and hypoxia inducible factors HIF 1, HIF 2 and CA IX in non-Hodgkin's lymphoma. AB - AIMS: Angiogenesis in solid tumour pathology is well established but less is known about its role in haematological malignancies. Our study investigated the immunohistochemical expression of a variety of angiogenic and hypoxic factors and microvessel densities on 110 cases of high- and low-grade non-Hodgkin's lymphomas and reactive lymphoid tissues. methods and results: Expression of vascular endothelial growth factor (VEGF) was present in 82 (96%) of the non-Hodgkin's cases and 35 (100%) of the reactive lymphoid tissue cases. Both hypoxia inducible factors 1 alpha and 2 alpha (HIF 1 alpha, 2 alpha) were weakly expressed in the majority of high- and low-grade lymphomas. Carbonic anhydrase IX (CA IX), a HIF inducible membrane-bound enzyme, expression was not abundant with membranous staining being present in seven (8%) of the lymphoma cases and none of the reactive cases. Thymidine phosphorylase (TP) was distributed amongst macrophages and follicular dendritic cells but was not present in the neoplastic population. The vasculature was stained using CD34 which gave rise to a distinct vascular, predominantly paracortical network present in low-grade lymphomas and reactive lymphoid tissue but which was lost in high-grade lymphomas. CONCLUSION: Our results suggest that non-Hodgkin's lymphomas may be less angiogenic and hypoxically driven than most solid tumours, which has implications for possible future therapies. PMID- 11895492 TI - Biliary epithelial trefoil peptide expression is increased in biliary diseases. AB - AIMS: Maintenance of the cellular integrity of the biliary epithelium may involve the production of mucins and mucin-associated peptides. In the luminal gastrointestinal tract, mucins and the mucin-associated trefoil peptides (TFF) are integral to cytoprotection and cellular repair of the mucosa. METHODS AND RESULTS: Samples of normal and diseased human liver tissue were examined using histological and immunohistochemical techniques, for the expression of TFF and mucins. Bile ducts were classified as small, medium or large depending upon the number of biliary epithelial cells. TFF expression was demonstrated in biliary epithelial cells of both normal and diseased liver tissue. TFF expression was greatest in the large bile ducts. In normal liver tissue, expression of at least one TFF was demonstrated in 2-7% of small bile ducts, 5-31% of medium bile ducts and 31-85% of large bile ducts. Seventy-seven percent of large bile ducts secreted mucins and all three TFF concurrently, compared with 3% of medium bile ducts and no small bile ducts. Biliary disease resulted in an increased expression of TFF1 and TFF3 in the medium bile ducts. CONCLUSIONS: The biliary epithelial cells in normal and diseased human liver tissue express TFF, particularly in the larger bile ducts. TFF expression may be up-regulated or induced in biliary diseases as a response to injury, as is seen in epithelial damage elsewhere in the gastrointestinal tract. PMID- 11895494 TI - Matrix metalloproteinase-2 expression correlates with morphological and immunohistochemical epithelial characteristics in synovial sarcoma. AB - AIMS: Synovial sarcoma is a unique mesenchymal tumour characterized by the presence of epithelial differentiation, although the mechanism involved in the epithelial morphology is still unclear. The aim of this study was to evaluate the function of matrix metalloproteinase-2 (MMP-2) in synovial sarcoma, in order to assess whether MMP-2 expression plays an important role in epithelial differentiation, or whether it contributes to a poor clinical outcome. METHODS AND RESULTS: Immunohistochemical stainings for MMP-2, cytokeratins (CKs) 7, 8, 18 and 19, and E-cadherin were performed for 58 (44 monophasic and 14 biphasic) cases of synovial sarcoma, and we compared the expression of these proteins with the histological and clinical findings. MMP-2 and E-cadherin expression was observed in 43 cases (74.1%) and in 18 cases (31.0%), respectively. Expression of these proteins was preferentially observed in the glandular components of biphasic tumours or the epithelioid areas of monophasic tumours. Statistically significant correlations were recognized between MMP-2 expression and E-cadherin expression of biphasic subtype. Moreover, there were statistically significant correlations between monophasic tumours with epithelioid areas and MMP-2 expression or E-cadherin expression. MMP-2 expression was correlated with epithelial differentiation as assessed by CK immunoreactivity. The expression of MMP-2 did not affect the overall survival rate in synovial sarcoma. CONCLUSIONS: MMP-2 expression seemed to have an important role to play in the epithelial differentiation of tumour cells in synovial sarcoma, through remodelling of the extracellular matrix and by changing the cytoskeletal interaction between the extracellular matrix and tumour cells. PMID- 11895493 TI - c-erbB-2 and c-Met expression relates to cholangiocarcinogenesis and progression of intrahepatic cholangiocarcinoma. AB - AIMS: The c-erbB-2 and c-Met proto-oncogenes are important for tumour invasiveness and metastasis in many types of malignant tumours. Previous studies have indicated that these proteins are associated with carcinogenesis in intrahepatic cholangiocarcinoma. In this study, we examined c-erbB-2 and c-Met expression by immunohistochemistry in hepatolithiasis, intrahepatic cholangiocarcinoma and metastatic lymph node, in order to clarify whether these proteins play a role in carcinogenesis and tumour metastasis in intrahepatic cholangiocarcinoma. METHODS AND RESULTS: In hepatolithiasis, the staining for c erbB-2 was positive in 14 of the 23 (61%) cases, while staining for c-Met was positive in eight of the 23 (35%) cases. In intrahepatic cholangiocarcinoma, staining for c-erbB-2 was positive in 45 of the 81 (55%) cases, while staining for c-Met was positive in 28 (35%) cases. The positivity of c-Met staining in intrahepatic cholangiocarcinoma was significantly higher in the differentiated type of cholangiocarcinoma than in the undifferentiated type. In addition, c-Met positive staining had an inverted correlation with tumour size, the presence of perineural invasion and the presence of lymph node metastasis. c-Met staining had a significantly higher positivity in cases at an early stage of intrahepatic cholangiocarcinoma. In contrast, the positivity of c-erbB-2 staining in intrahepatic cholangiocarcinoma was significantly higher in cases with lymph node metastasis than in cases without. In metastatic lymph nodes, the staining for c erbB-2 was positive in 20 of the 25 (80%) cases, while staining for c-Met was positive in six of the 25 (24%) cases. There was no difference in survival between c-erbB-2-positive and negative patients. However, the patients with c-Met positive tumours had a significantly longer survival than those with c-Met negative tumours in the medium survival term. The multivariate analysis showed the presence of lymph node metastasis, lymphatic permeation and histological differentiation to be independent prognostic factors. CONCLUSION: These results indicate that increased c-Met expression participates in cholangiocarcinogenesis and in the early developmental stages of intrahepatic cholangiocarcinoma, while increased c-erbB-2 expression contributes to the development of cholangiocarcinogenesis into an advanced stage associated with tumour metastasis. PMID- 11895495 TI - Primary monotypic epithelioid angiomyolipoma of bone. AB - AIMS: Monotypic epithelioid angiomyolipoma is a distinct and definable variant of angiomyolipoma, composed of monomorphous epithelioid cells that show HMB45 immunoreactivity. Angiomyolipoma, including its morphological variants, belongs to the family of perivascular epithelioid cell tumour. METHODS AND RESULTS: The tumour was examined using immunohistochemical staining and by transmission electron microscopy. Neoplastic cells showed a cytoplasmic granular positivity for HMB45. CONCLUSIONS: Extrarenal angiomyolipomas are rare and, to the best of our knowledge, this is the first reported case of a primary monotypic epithelioid angiomyolipoma of bone in a patient without evidence of tuberous sclerosis. PMID- 11895496 TI - Sclerosing haemangioma of the lung is positive for MIB-1 in cell membrane and cytoplasmic staining pattern. AB - AIMS: MIB-1 is a monoclonal antibody raised against the recombinant part of the Ki67 antigen, which is expressed in the nuclei of all the active part of the cell cycle. Recently, cell membrane and cytoplasmic pattern of staining with MIB-1 in hyalinizing trabecular adenoma of the thyroid was reported. Sclerosing haemangioma of the lung is a tumour of uncertain histogenesis with a usually benign course. The tumour is mainly composed of cuboidal cells that line the papillary or tubular structure and pale cells that are found in the stroma. METHODS AND RESULTS: We examined a cell membrane and cytoplasmic immunoreactive pattern for MIB-1 antibody in sclerosing haemangioma. In all of the five cases examined, distinct cell membrane and cytoplasmic MIB-1 positivity mainly in the pale cells was noted. None of the carcinomas of the lung showed this type of staining with MIB-1. CONCLUSION: This result shows that, although the staining may be a result of cross reaction, cell membrane and cytoplasmic MIB-1 positivity is unique in that it could distinguish sclerosing haemangioma from other malignant epithelial tumours of the lung. PMID- 11895497 TI - Creating digital images of pathology specimens by using a flatbed scanner. PMID- 11895498 TI - Somatic hypermutation of immunoglobulin heavy chain variable region genes in thymic marginal zone B-cell lymphoma of MALT type of a patient with Sjogren's syndrome. PMID- 11895499 TI - Atypical cellular perineurioma. PMID- 11895500 TI - Nodular histiocytic/mesothelial hyperplasia: a process mediated by adhesion molecules? PMID- 11895501 TI - Pulmonary metastases from a chromophobe renal cell carcinoma: 10 years' evolution. PMID- 11895502 TI - Adenocarcinoma arising within a Crohn's-related anorectal fistula: a form of anal gland carcinoma? PMID- 11895506 TI - Dermatology practice in primary health care services: where do we stand in the Middle East? AB - BACKGROUND: There has been a distinct expansion of the primary health care services in the Middle East over the past two decades. As a consequence, the exposure of primary care physicians (PCPs) to skin disorders has increased. However, information is lacking regarding the level of proficiency of PCPs in this field. OBJECTIVE: The purpose of our study is to assess the ability of the primary care physicians, with or without training in dermatology, to identify, diagnose and manage skin disorders. MATERIAL AND METHODS: Physicians at university-hospital primary-care clinics were asked to answer a multiple-choice questionnaire regarding various dermatoses. These were grouped into: common, infrequent and rare. Questions included identification of the correct description of the skin lesion, diagnosis, treatment and the desirability of referral. Demographic characteristics of the physicians were also assessed. RESULTS: Nineteen PCPs were included. The eight PCPs who had had specific training in dermatology showed performance superior to that of the PCPs who did not (P = 0.04). Not surprisingly, PCPs were able to make the correct diagnosis more frequently for the common dermatoses than for the infrequent or rare dermatoses (P = 0.001). On the other hand, when asked to recognize a correct description of the skin lesion, the PCPs were most often correct with rare dermatoses, and least often correct with common dermatoses (P = 0.04). CONCLUSION: PCPs with a short period of specific clinical training in dermatology perform better in identifying, diagnosing and managing skin disorders than those without. Such training for PCPs should be considered to provide more effective delivery of health care. PMID- 11895507 TI - Cicatricial alopecia; a dermatopathologic and immunopathologic study of 33 patients (pseudopelade of Brocq is not a specific clinico-pathologic entity). AB - BACKGROUND: Pseudopelade of Brocq (PB) is a permanent progressive scarring alopecia characterized by numerous alopecic patches localized only in the scalp, that tend to coalesce into larger, irregular plaques with policyclic borders. PB can be considered either the final atrophic stage of several scarring disorders such as lichen planus pilaris (LPP) and discoid lupus erythematosus (DLE) (secondary PB) or an autonomous disease (primary PB). The aim of this study was to assess the incidence of primary vs. secondary PB by a combined histopathological and immunopathological study in a series of patients who fulfilled the clinical diagnostic criteria for PB set forth by Braun Falco et al. METHODS: We studied 33 patients (5 males and 28 females, whose age ranged from 24 to 75 years). The duration of the disease (from onset to biopsy) ranged from 3 months to 8 years. Serum samples were tested for circulating auto-antibodies (antinuclear antibodies anti ENA, anticentromere, anti-Scl70, antithyroid, antigastric parietal cells) circulating immune complexes, total and single fraction (C3, C4) complement activity. The skin biopsies taken from the active advancing margin of the more recent alopecic patch were bisected vertically, one was sent for histopathological examination, and the other for the immunofluorescence studies. RESULTS: In all patients the serum tests above were found to be negative or normal. Histopathologically, 11 biopsies (33.3%) displayed findings typical for LPP whereas seven cases (21.2%) showed typical DLE features. In the remaining 15 cases (45.5%) histopathological findings were not suggestive of any specific dermatosis. DIF investigations showed findings typical of LPP in six cases (18.1%) and typical of DLE in seven cases (21%). In three cases we did not find findings typical of LPP, DLE, or any other specific dermatitis. In 11 cases no immunological deposits could be detected and therefore were classified as negative. CONCLUSION: In conclusion, PB is a type of scarring alopecia of the scalp associated with a peculiar clinical presentation and evolution, which cannot be considered an autonomous nosologic entity because in 66.6% of patients it is the end stage of other inflammatory chronic diseases such as LPP and DLE. It is conceivable that even in those cases in which the histopathological and immunopathological findings did not allow for a specific diagnosis, LPP and DLE were also involved. It is noteworthy that in our study the histopathological and the immunopathological examinations did not conflict and often the results were even coincidental, thus confirming the compatibility of the combined histo-immunopathological approach in the diagnostic evaluation of PB. PMID- 11895508 TI - Subcutaneous pheohyphomycosis in India--a case report and review. AB - BACKGROUND: Pheohyphomycosis is a rare infection caused by dematiaceous fungi, affecting the skin and subcutis, paranasal sinuses, or central nervous system. METHODS: A case of subcutaneous pheohyphomycosis in the lumbar region is reported. The Indian literature on subcutaneous pheohyphomycosis is also reviewed. RESULTS: In India, 23 patients with subcutaneous pheohyphomycosis have been reported, distributed throughout the country in a belt from north to south, sparing the eastern and western regions. The age of the patients ranged from 3 to 60 years, with a male to female ratio of 1.3 : 1. A relatively early age of onset was observed. A history of prior injury was recalled by five patients. The lower extremities were involved in eight cases, upper extremities in five, gluteal region in two, lumbar area and submandibular area in one, face in two and disseminated disease was seen in four cases. Three of these cased died during follow up. Osteomyelitis was observed in two cases, hepatosplenomegaly in one, and lymph node involvement in two. Carcinomatous change developed in a long standing lesion of 33 years. Thirteen species from seven genera of dematiaceous fungi were isolated. Phialophora dermatidis was the most common isolate. CONCLUSIONS: Indian patients with subcutaneous pheohyphomycosis are afflicted at an earlier age. Exophiala dermatidis seems to be associated with more fatalities. Treatment with newer azoles seems promising, and excision alone or combined with azoles is a good therapeutic modality. PMID- 11895509 TI - Cigarette smoking associated with premature facial wrinkling: image analysis of facial skin replicas. AB - BACKGROUND: Despite the obvious relation between smoking and facial wrinkling, grossly undetectable wrinkling and the consequences of smoking on the face have been poorly studied. OBJECTIVE: To assess the risk factor of cigarette smoking on the development of premature facial wrinkling. METHODS: One hundred and twenty three nonsmokers, 160 current smokers, and 67 past smokers, aged 20-69 years, were studied. Cigarette smoking status, weight changes, average sun exposure time (recreational and occupational) in 1 month, and past medical and facial cosmetic surgery were quantified by self-questionnaire. Computerized image analysis of silicone skin replicas was used in addition to clinical visual measurement, and a severity score based on predetermined criteria was assigned to each patient. RESULTS: Current smokers have a higher degree of facial wrinkling than nonsmokers and past smokers. Past smokers who smoked heavily at a younger age show less facial wrinkling than current smokers. In the analysis, which was adjusted for age group, the relative risk of moderate to severe wrinkling for current smokers compared with nonsmokers was 2.72 (confidence interval, CI: 1.32-3.21, P < 0.05). In current smokers, the relative risks associated with more than 19 pack-years and 11-19 pack-years of smoking compared with nonsmokers were 2.93 (CI: 1.14-4.1, P < 0.05) and 1.75 (CI: 1.54-3.67, P < 0.05), respectively. On image analysis of facial skin replicas, the mean values of Ra (arithmetic average roughness), Rz (average roughness), and Rt (distance between the highest and lowest values) of current smokers were higher than those of nonsmokers and past smokers in all age groups. This indicates a strong correlation between cigarette smoking and skin wrinkling. In addition, microscopic superficial wrinkling (Ra and Rt) was noted in current smokers in the younger age group (20-39 years). CONCLUSIONS: This study suggests that attention should be paid to smoking-associated facial wrinkling (not evident from a visual assessment) in young people and added to the list of disorders seemingly caused by smoking. PMID- 11895511 TI - Cutaneous leishmaniasis in Sanliurfa: epidemiologic and clinical features of the last four years (1997-2000). AB - BACKGROUND: Sanliurfa is located in south-east Anatolia, the region with the largest focus of anthroponotic cutaneous leishmaniasis (CL) in Turkey. The present study was designed to determine the epidemiological and clinical patterns of cases of CL in the Sanliurfa area over a period of 4 years (1997-2000). METHODS: 2120 CL cases attending the Harrankapi in this study. RESULTS: There were 790 cases of CL in 1997, and 778 in 1998. The numbers declined to 277 in 1999 and to 275 in 2000. The majority of patients (70%) were less than 20 years of age, with the highest percentage (27%) occurring in the 5- to 9-year age group. Acute CL and chronic CL were observed in 1926 (90.8%) and 63 (3%) patients, respectively. Leishmaniasis recidivans (LR) was found only in 131 (6.2%) patients. Lesions of CL were seen mainly on the exposed parts of the body such as the face and neck (57.5%), upper limbs (32.2%) and lower limbs (10.2%). Ninety-three percent of the patients were residents of central Sanliurfa from the southern and eastern parts of the city, areas with poor housing and low socio economic conditions. CONCLUSION: Although the incidence of CL showed a sharp decline from 1997 to 2000 in Sanliurfa, this does not mean that the disease is on its way to eradication. PMID- 11895510 TI - Scrofuloderma and Sweet's syndrome. AB - BACKGROUND: In recent years, the rare association of Sweet's syndrome with nontuberculous mycobacterial lymphadenitis has been reported. OBJECTIVE: To report the clinical, demographic, and bacteriologic data and association with Sweet's syndrome of 18 patients with scrofuloderma and scrofuloderma-like condition caused by nontuberculous mycobacterial infections seen during the past 7 years (1994-2000). METHODS: In all patients, a biopsy specimen was obtained for histopathologic and microbiologic studies. Patients from whom Mycobacterium tuberculosis and nontuberculous mycobacteria were isolated from the culture of skin biopsy specimens were included. Deep fungal infection was excluded by the lack of a fungal element in histologic section and cultural methods. The patients were treated with antimicrobials or antituberculous drugs according to the causative species. RESULTS: Eighteen cases of scrofuloderma (nine male, nine female; mean age, 36.9 years) were found among 104 patients with cutaneous tuberculosis and nontuberculous mycobacterial cutaneous infections. Sixteen of the 18 cases had lymphadenitis as the underlying focus of scrofuloderma: 15 cases occurred in the cervical group and one case in the inguinal area. One case drained from the soft tissue and one from the paranasal air sinus. Five cases had multiple episodes of Sweet' s syndrome during the course of treatment. Most cases in this group (four of the five) were middle-aged women with cervical lymphadenitis, and the most common species were rapid growers. CONCLUSIONS: Age, sex, and the site of infection may have some influence on the association with Sweet's syndrome in nontuberculous mycobacterial infections. PMID- 11895512 TI - The therapeutic potential of calcipotriol in diseases other than psoriasis. AB - BACKGROUND: Calcipotriol is a vitamin D analog, which has antiproliferative and anti- inflammatory effects, and stimulates terminal differentiation. It has been an established treatment for psoriasis since 1991 in Europe and 1994 in the USA. OBJECTIVE: To assess the clinical efficacy of topical calcipotriol in diseases other than psoriasis. RESULTS: A total of 36 original papers were found describing 21 different diseases in which the clinical use of calcipotriol ointment was described as having an effect. These papers were predominantly case reports (n = 22) and observational studies (n = 5), but nine papers described small randomized controlled trials. CONCLUSIONS: Calcipotriol may generally be effective in diseases characterized by pathogenic elements, such as impaired differentiation or increased keratinocyte proliferation, and activated T lymphocytes. This study of the existing literature suggests that randomized, double-blind, placebo-controlled studies of calcipotriol therapy may be worthwhile in a number of diseases. PMID- 11895513 TI - Lichenoid nail changes as sole external manifestation of graft vs. host disease. AB - A 56-year-old-man who had refractory anemia with an excess of blasts underwent an allogeneic peripheral blood stem cell transplantation (PBSCT) from his brother after preparation with melphalan and fludarabin. He received GvHD (graft-vs.-host disease) prophylaxis with cyclosporine from day -1 at a daily dose of 5 mg/kg of body weight. The daily dosage was tapered gradually from day +20. On post-PBSCT day 68 he developed acute cutaneous GvHD grade 3 and acute gastrointestinal GvHD grade 2-3, which was resolved with a daily dose of 1 mg/kg of body weight of prednisone. The patient was discharged in good clinical condition and without signs of GvHD, and he started tapering his immunosuppressive treatment. By day 160 he developed oral lichen planus-like changes, with several reticulate white lesions on the oral mucosa. A biopsy specimen was microscopically consistent with lichenoid GvHD (Fig. 1). By day 150 after PBSCT, when he was being treated with CsA 100 mg once daily and prednisone 10 mg once daily, his fingernails started to grow abnormally and gradually became dystrophic and painful. Two months later his toenails became similarly affected. Although affecting all finger and toe nails, the lesions were especially important in both thumbs. Physical examination revealed multiple findings on his nails (Fig. 2): thickening, fragility, onycholysis, longitudinal striations, and even pterygium. The micological cultures were negative. A biopsy specimen showed an sparse papillary dermis lymphoid infiltrate with focal exocytosis and presence of isolated multiple necrotic keratinocytes (Fig. 3). These findings were interpreted as a lichenoid GvHD with oral and nail involvement. The patient did not have other associated cutaneous lesions. He did not develop signs or symptoms consistent with hepatic GvHD. In May 2000 thalidomide was added to the immunosuppressive therapy, at a daily dose from 100 to 300 mg according to tolerance (constipation, sedation, ...). The lesions on the oral mucous showed a substantial improvement, but the nail changes remained more or less stable. Thalidomide was discontinued after 7 months because the patient displayed numbness and tingling in the hands and feet consistent with a peripheral neuropathy. Twenty days later he stopped taking thalidomide and the oral lichenoid lesions worsened, resulting in difficulty in eating. He also developed periungueal erythema, swelling and intense pain after minimal trauma. The daily dose of prednisone increased to 20-30 mg with moderate improvement. However, the dose could not be increased because of the secondary immunosuppressive effects. Twenty-three months post-PBSCT the patient remains with intense oral and nail lichenoid lesions. PMID- 11895515 TI - Cutaneous ciliated cyst on the cheek in a male. PMID- 11895514 TI - Prolidase deficiency. PMID- 11895516 TI - Stingray injury in a domestic aquarium. PMID- 11895517 TI - Regression of an acral lentiginous melanoma with an immunotherapy using a Mycobacterium tuberculosis-extracted polysaccharide complex (Tubercin). PMID- 11895518 TI - Three cases of Nattrassia mangiferae (Scytalidium dimidiatum) infection in Singapore. PMID- 11895519 TI - Localized linear IgA disease responding to colchicine. PMID- 11895523 TI - How to make your article more readable. PMID- 11895526 TI - Balancing involvement: employees' experiences of merging hospitals in Sweden. AB - BACKGROUND: The health care of today stands in front of demands on financial and structural changes. New technology and global economy are forces driving on the change process. AIMS: The aim of this study is to describe and broaden the understanding of the employees' experience of being involved in a merger between two health care districts in Sweden. METHODS: This study was carried out from a qualitative approach according to the grounded theory tradition. From a theme guide with specific questions, 31 interviews were carried out with employees working in the health care. FINDINGS: Five categories emerged from the body of interviews: balancing involvement, trust respect, challenge and commitment. Balancing involvement was defined as an overall core category related to the other categories. The categories trust, respect, challenge and commitment were related to subcategories and affected the core category balancing involvement. CONCLUSIONS: The overall findings point to the importance of balancing the employees' involvement in order to reach goal fulfilment change in a merger process. PMID- 11895527 TI - From caring to managing and beyond: an examination of the head nurse's role. AB - AIM: The aim of this study was to depict the essence of what head nurses do, and how they perform their managerial role. METHODS: To achieve this, the work behaviour of 48 head nurses was examined by a semi-structured observation technique for 6 hours each. RESULTS: Results demonstrated that head nurses spent a large proportion of their time in clinical practice, followed by co-ordinating care, operating the unit's functions, and leading staff. Personnel management and quality improvement occupied only a marginal share of head nurses' time. CONCLUSIONS: These results implied that head nurses exhibited a management style orientated to maintenance rather than to re-creation, focusing more on the 'doing' and the 'here and now' aspects of the job than on leading, planning, and proactive problem solving. PMID- 11895529 TI - The psychological well-being of renal peer support volunteers. AB - AIM OF THE STUDY: The purpose of this study was to describe the characteristics of renal peer support volunteers (PSVs) and explore the effects on their psychological well-being from helping others. BACKGROUND: Dialysis patients, transplant patients and family members who become renal PSVs receive special training in empathy, listening, self-awareness and problem solving. The trained renal PSVs offer a unique service to others struggling to learn to live with renal failure because they have faced the same struggles. METHODS: This exploratory study utilized a longitudinal design. The first time for data collection was immediately after the volunteers had completed a Kidney Foundation of Canada training programme. Subsequent interviews were at time intervals of 4, 8 and 12 months after the first interview. Information on the psychological well being of the volunteers was collected at each interview in two different ways: the 38-item Mental Health Inventory (MHI) and open-ended questions. FINDINGS: Thirty-one PSVs completed all four interviews. The average age of the volunteers was 45 years and almost half had a university level of education. They identified themselves as belonging to 12 different ethno-cultural groups. Analysis of the quantitative data from the MHI indicated that the mental health of the PSVs stayed remarkably stable over time. Analysis of the qualitative data from the open-ended questions revealed four major themes which, taken together, showed notable increases in personal growth and well-being for the PSVs over time. CONCLUSION: After participating in a training programme, renal PSVs maintained, and possibly improved, their own well-being by helping others with chronic renal failure. PMID- 11895528 TI - An empirical test of the Nursing Role Effectiveness Model. AB - AIMS OF THE STUDY: This study investigated the propositions depicted in the Nursing Role Effectiveness Model, in which nurse and patient structural variables were expected to influence nurses' role performance, which, in turn was expected to affect patient outcome achievement. RATIONALE/BACKGROUND: Increasingly, nurses are expected to demonstrate their contribution to patient outcome achievement as a basis for evaluating practice and for monitoring improvements in practice. A model was developed that describes nursing practice in relationship to the roles nurses assume in health care, and links patient and system outcomes to nurses' role functions (Nursing Economics 1998: 16, 58-64, 87). RESEARCH METHODS: A cross sectional design was used to collect data on the structure, process, and outcome variables. Data were collected through structured questionnaires and chart audit, involving a total of 372 patients and 254 nurses from 26 general medical-surgical units in a tertiary care hospital. Patient structural variables included medical diagnosis, age, gender and education. Nurse structural variables included educational preparation and length of hospital experience. The unit structural variables included the adequacy of time to provide care, autonomy, and role tension. The quality of nurses' independent role performance was assessed by collecting data from patients on their perception of the quality of nursing care. Nurses' interdependent role performance was assessed by collecting data from nurses on the quality of nurse communication and co-ordination of care. Patient outcomes were assessed through self-report and consisted of the patients' therapeutic self-care ability, functional status, and mood disturbance at the time of hospital discharge. Structural equation modelling was used to test the hypothesized relationships among the structural, process, and outcome variables. RESULTS: Patients viewed nurses' independent role performance more effective on units where nurses reported less autonomy but more time to provide care. The quality of nurse communication was higher on units where nurses had higher education, more autonomy, less hospital experience, and lower role tension. However, the co-ordination of care was more effective on units where nurses had higher education, greater hospital experience, less autonomy and role tension. The three role performance variables were associated with patients' therapeutic self-care ability at hospital discharge. Nurses' independent role performance was associated with better patient functional status and less mood disturbance at hospital discharge. The role performance variables fully mediated the effect of the structural variables on patient outcomes, lending support for the proposition that nurses' role performance explains the relationship between structural variables, such as nurse education and autonomy, and patient outcome achievement. DISCUSSION: The Nursing Role Effectiveness Model provides a well-defined conceptual framework to guide the evaluation of outcomes of nursing care. For the most part the hypothesized relationships among the variables were supported. However, further work is needed to develop an understanding of how nurses engage in their co-ordinating role functions and how we can measures these role activities. PMID- 11895531 TI - Measurement of outpatients' views of service quality in a Finnish university hospital. AB - AIMS OF THE STUDY: To describe the development of service quality in the surgical and medical outpatient departments of a university hospital. RATIONALE: Measurement and documentation of service quality and the use of these data as a basis for improvement are prerequisites for service improvement. METHODS: A survey conducted over a 3-week period using a 12-item questionnaire measuring service quality on a rating scale from 4 to 10 (extremely poor - excellent quality). Nineteen outpatient departments participated at a Finnish university hospital, 7679 voluntary patients visiting the outpatient departments between 1997 and 1999 completed the questionnaires. RESULTS: The questionnaire, which has not been described in use previously, proved to be a good tool enabling systematic access to patient feedback on service quality in outpatient departments. The instrument allowed the detection of strengths of the service provided by a large organization and long-term trends from patients' perspective. Patient evaluations were mainly good and the results improved year by year, although the instrument was built on questions concerning issues that were rated poorest by patients. The poorest ratings were related to access to information and adherence to appointment times. CONCLUSIONS: The service quality instrument offers a means to generate information to be used for service quality improvement in outpatient departments. PMID- 11895530 TI - Promoting a good life among people with Alzheimer's disease. AB - AIM: To illuminate the meaning of offering care and a place to live to people with Alzheimer's disease in a special care unit. RATIONALE: There is a need to gain a deeper understanding about so called 'homelike' care settings, and about how to promote experiences of being at home in residents with Alzheimer's disease. The study is part of a long-term study in a special care unit. METHODS: The study comprises phenomenological hermeneutic interpretation of interviews with 10 care providers. RESULTS: The analyses revealed a number of caring aspects such as, for example, 'viewing dignity and striving to preserve a sense of self in the resident', 'encouraging a sense of belonging', 'offering relief' and 'promoting a sense of power and control in the resident', although integrated and reflected in each other. The caring aspects constituted the themes confirmation, familiarity, communion and agency considered as dimensions of the good life. CONCLUSION: To avoid simplification in which, for example, the furniture from a certain decade become the standard for good care, it seems important to focus upon the meaning of the good life. Care that promotes a good life of people with Alzheimer's disease seemed relationship centred. PMID- 11895532 TI - Effects of massage on pain and anxiety during labour: a randomized controlled trial in Taiwan. AB - AIMS: To investigate the effects of massage on pain reaction and anxiety during labour. BACKGROUND: Labour pain is a challenging issue for nurses designing intervention protocols. Massage is an ancient technique that has been widely employed during labour, however, relatively little study has been undertaken examining the effects of massage on women in labour. METHODS: A randomized controlled study was conducted between September 1999 and January 2000. Sixty primiparous women expected to have a normal childbirth at a regional hospital in southern Taiwan were randomly assigned to either the experimental (n=30) or the control (n=30) group. The experimental group received massage intervention whereas the control group did not. The nurse-rated present behavioural intensity (PBI) was used as a measure of labour pain. Anxiety was measured with the visual analogue scale for anxiety (VASA). The intensity of pain and anxiety between the two groups was compared in the latent phase (cervix dilated 3-4 cm), active phase (5-7 cm) and transitional phase (8-10 cm). RESULTS: In both groups, there was a relatively steady increase in pain intensity and anxiety level as labour progressed. A t-test demonstrated that the experimental group had significantly lower pain reactions in the latent, active and transitional phases. Anxiety levels were only significantly different between the two groups in the latent phase. Twenty-six of the 30 (87%) experimental group subjects reported that massage was helpful, providing pain relief and psychological support during labour. CONCLUSIONS: Findings suggest that massage is a cost-effective nursing intervention that can decrease pain and anxiety during labour, and partners' participation in massage can positively influence the quality of women's birth experiences. PMID- 11895533 TI - Chinese women's perceptions of the effectiveness of antenatal education in the preparation for motherhood. AB - AIM OF THE STUDY: This was an exploratory descriptive study using mixed methodology to investigate Hong Kong Chinese women's perceptions of the effectiveness of antenatal education in their preparation for motherhood. DESIGN: In the first phase, the structure and process of five antenatal classes on the topic of motherhood were observed using an observation guide. In the second phase 11 women who had attended the antenatal classes were interviewed in two focus groups, using a semi-structured interview guide. FINDINGS: In respect to the structure of the classes women revealed that large class sizes and the didactic mode of teaching inhibited learning. While they were satisfied with the date and time of antenatal classes, and the information about self and baby care being provided, they felt unprepared for the demands of motherhood. Further themes identified from the analysis were: anticipating personal needs for antenatal preparation for motherhood, unrealistic preparation for breastfeeding problems, inadequate preparation for baby care, unfulfilled informational needs and conflicting advice from antenatal educators. CONCLUSION: The conclusion highlights Chinese culturally specific changes needed in the content and mode of antenatal education. In addition, recommendations are made for antenatal educators to work within a framework of adult Chinese learning styles in order to meet the educational needs of Chinese women. PMID- 11895534 TI - Nursing the clinic, being there and hovering: ways of caring in a British fertility unit. AB - AIM: To explore the meaning of caring in a fertility unit and to present the findings and discuss their implications for practice. BACKGROUND: Little research has been published internationally on the nature of nursing care in the fertility field. This study was intended to stimulate debate over the nature of care in fertility work. METHODOLOGY: An ethnographic approach, which included part-time participant observation and in-depth focused interviews with staff and patients. Data were also collected using a field and a research diary. Data were analysed using a modified thematic analysis. FINDINGS: The data suggest that caring is strongly linked with nurses and that patients' expectations of nurses may be more practically than emotionally focused. I argue that the organization of nursing work, which I call 'nursing the clinic and the doctor', facilitated this practical approach to caring. I discuss two features of nursing the clinic and the doctor: hovering and being there. CONCLUSIONS: While there may be other factors that influence the practice of this form of non-intimate caring, nursing in this way may be what patients desire and may be congruent with managing emotions. The findings have implications for the discourse on intimacy and caring within fertility nursing as well as in different outpatient settings. PMID- 11895535 TI - Getting evidence into practice: the meaning of 'context'. AB - AIM OF PAPER: This paper presents the findings of a concept analysis of 'context' in relation to the successful implementation of evidence into practice. BACKGROUND: In 1998, a conceptual framework was developed that represented the interplay and interdependence of the many factors influencing the uptake of evidence into practice [Kitson A., Harvey G. & McCormack B. (1998) Quality in Health Care 7, 149]. One of the key elements of the framework was 'context', that is, the setting in which evidence is implemented. It was proposed that key factors in the context of health care practice had a significant impact on the implementation and uptake of evidence. As part of the on-going development and refinement of the framework, the elements within it have undergone a concept analysis in order to provide some theoretical and conceptual rigour to its content. METHODS: Morse's [Morse J.M. (1995) Advances in Nursing Science 17, 31; Morse J.M., Hupcey J.E. & Mitcham C. (1996) Scholarly Inquiry for Nursing Practice. An International Journal 10, 253] approach to concept analysis was used as a framework to review semi-nal texts critically and the supporting research literature in order to establish the conceptual clarity and maturity of 'context' in relation to its importance in the implementation of evidence-based practice. FINDINGS: Characteristics of the concept of context in terms of organizational culture, leadership and measurement are outlined. A main finding is that context specifically means 'the setting in which practice takes place', but that the term itself does little to reflect the complexity of the concept. Whilst the themes of culture and leadership are central characteristics of the concept, the theme of 'measurement' is better articulated through the broader term of 'evaluation'. CONCLUSIONS: There is inconsistency in the use of the term and this has an impact on claims of its importance. The concept of context lacks clarity because of the many issues that impact on the way it is characterized. Additionally, there is limited understanding of the consequences of working with different contexts. Thus, the implications of using context as a variable in research studies exploring research implementation are as yet largely unknown. The concept of context is partially developed but in need of further delineation and comparison. PMID- 11895536 TI - The effects of subgingival calculus on the clinical outcomes of locally-delivered controlled-release doxycycline compared to scaling and root planing. AB - BACKGROUND/AIM: The effect of subgingival calculus on the clinical outcomes of the local delivery of antimicrobials is unknown. This study examines the clinical outcomes of treatment with locally delivered controlled-release doxycycline (DH) or scaling and root planing (SRP) in subsets of adult periodontitis patients with known baseline levels of subgingival calculus. METHODS: The data examined were obtained from 393 patients who participated in 2 multi-center trials. All patients had baseline subgingival calculus levels assessed and were then treated at baseline and month 4 with either DH or SRP. Clinical attachment levels (CAL), pocket depth (PD) and bleeding on probing (BOP) were assessed at baseline and months 1, 2, 4, 5, 6, 8 and 9. RESULTS: Treatment with either DH or SRP resulted in significant statistical and clinical improvements in CAL, PD and BOP. These clinical outcomes were equivalent regardless of the extent of subgingival calculus present at baseline. CONCLUSIONS: The results indicate that the primary clinical effects of these therapies are the result of a disruption and reduction of the subgingival plaque and not the effect of the removal of subgingival calculus and contaminated cementum. PMID- 11895537 TI - A laser Doppler study of gingival blood flow variations following periosteal stimulation. AB - OBJECTIVES: Evaluation of the modifications occurring in human gingival blood flow following periosteal stimulation. MATERIAL AND METHODS: Laser Doppler was used to measure the gingival blood flow (GBF). The reproducibility of the technique was validated by comparing measures made at intervals of 1 week. Sensitivity was verified by recording GBF before and after injection of an anesthetic containing a vasoconstrictor. Finally, 12 patients were subjected to GBF measurements before and 8 days after periosteal stimulation prior to gingival grafting. RESULTS: The laser Doppler accurately measured GBF. Measurements made at day 0 and day 7 were not statistically different (p=0.60). After injection of an anesthesic solution containing vasoconstrictor, the laser Doppler recorded a sharp decrease of the GBF (p=0.04). The patient that underwent periosteal stimulation showed statistically significant increases (p=0.02) in GBF before and 1 week post-stimulation. CONCLUSION: Periosteal stimulation induces significant increases in the GBF after 1 week. PMID- 11895539 TI - IL-8 and TNF-alpha from peripheral neutrophils and acute-phase proteins in periodontitis. AB - OBJECTIVE: In other studies, we have found deviant functions in peripheral neutrophils in periodontitis. The aim here was to study (1) the release of cytokines, IL-8 and TNFalpha, from neutrophils in 15 treated periodontitis patients and pair-matched controls as well as (2) the effects of cigarette smoking. MATERIAL AND METHODS: Cytokines released in the incubation medium from un-stimulated and Fcgamma-R-stimulated neutrophils and some acute-phase reactants were measured with ELISA. RESULTS: Non-smoking patients had trends for lower TNFalpha release compared to non-smoking controls, while corresponding trends were rather similar for Il-8. Smoking had a moderate but inconsistent effect on the release of both cytokines. However, in patients, the ratio between stimulated/un-stimulated release of Il-8 was significantly lowered by smoking (p<0.03). The parameters of inflammation in plasma differed only slightly between patients and controls, indicating that periodontal disease in a quiet phase has a negligible systemic effect with the possible exception for a higher IL-8 level. In contrast, smoking had significant systemic effect on the neutrophil count and IgG levels. CONCLUSIONS: Release of IL-8 and TNF-alpha from peripheral neutrophils and various parameters of inflammation in plasma seem to be affected more by cigarette smoking than periodontal disease. PMID- 11895538 TI - Non-surgical periodontal therapy with adjunctive topical doxycycline: a double blind randomized controlled multicenter study. AB - AIM: Evaluation of the clinical effect of topical application of doxycycline adjunctive to non-surgical periodontal therapy. METHODS: A total of 111 patients suffering from untreated or recurrent moderate to severe periodontitis at 3 different centers (Heidelberg, Frankfurt, Nijmegen) were treated in this double blind split-mouth study. In each patient, 3 different treatment modalities were assigned randomly to 3 test teeth: scaling and root planing alone (SRP), SRP with subgingival vehicle control (VEH), and SRP with subgingival application of a newly developed biodegradable 15% doxycycline gel (DOXI). At baseline, clinical parameters were measured at all single rooted teeth using a reference splint: PlI, PPD, relative attachment level (RAL-V), GI. 3 strata were generated according to baseline PPD: (i) 5-6 mm, (ii) 7-8 mm, (iii) > or =9 mm. Not more than 50% active smokers were allowed to each stratum. 3 and 6 months after therapy re-examination was performed by examiners blinded to baseline data and test sites. The statistical comparison of RAL-V gain and PPD reduction between the treatments was based on a repeated measures ANOVA with correction according to Huynh & Feldt. The comparison of SRP versus DOXI was considered as the main study question. RESULTS: 110 patients finished the 3 months and 108 the 6 months examination. The study did not show adverse effects of VEH or DOXI except for one singular inflammation that occurred 2 months after application of the doxycycline gel. DOXI provided statistically significantly more favorable PPD reduction (SRP: -2.4+/-1.4 mm, VEH: -2.7+/-1.6 mm, DOXI: -3.1+/-1.2 mm; SRP versus DOXI p=0.0001, VEH versus DOXI p=0.0066) and RAL-V gain (SRP: 1.6+/-1.9 mm, VEH: 1.6+/-2.2 mm, DOXI: 2.0+/-1.7 mm; SRP versus DOXI p=0.027, VEH versus DOXI p=0.038) than SRP and VEH after 6 months. CONCLUSIONS: Adjunctive topical subgingival application of a biodegradable 15% doxycycline gel was safe and provided more favorable RAL-V gain and PPD reduction than SRP alone and VEH. Thus, by use of topical doxycycline the threshold for surgical periodontal therapy might be moved toward deeper pockets. PMID- 11895540 TI - Gingival recession in smokers and non-smokers with minimal periodontal disease. AB - BACKGROUND/AIMS: Smoking is a major risk factor for destructive periodontal disease. There is limited information with regard to effects of smoking in subjects with minimal periodontal destruction. The aim of the present investigation was to assess the development of gingival recession in young adult smokers and non-smokers. METHODS: 61 systemically healthy young adults, 19 to 30 years of age completed the final examination. 30 volunteers smoked at least 20 cigarettes per day, whereas 31 subjects were non-smokers. Clinical periodontal conditions were assessed 4x within a time period of 6 months. Site-specific analyses considering the correlated structure of data were performed. RESULTS: At the outset, 50% of subjects presented with gingival recession at 1 or more sites. There was no significant difference in the prevalence of gingival recession between non-smokers and smokers. Severe recession in excess of 2 mm affected about 23% non-smokers but only 7% smokers. Some further gingival recession developed during the 6-month observation period. In a multivariate logistic regression analysis, the risk for recession development appeared not to be influenced by smoking status after adjusting for periodontal probing depth, recession at baseline, tooth brushing frequency, gender, jaw, tooth type and site. CONCLUSIONS: Present data did not support the hypothesis that smokers are at an increased risk for the development of gingival recession. PMID- 11895541 TI - Clinical, radiographic and biochemical assessment of IL-1/TNF-alpha antagonist inhibition of bone loss in experimental periodontitis. AB - OBJECTIVES: To assess clinical, radiographic, and biochemical markers as diagnostic indicators of disease activity by comparing ligature-induced bone loss in the presence or absence of IL-1/TNF-alpha antagonist inhibition of bone loss in a primate model. MATERIAL AND METHODS: 6 animals with a naturally-occurring gingivitis were evaluated over a 6-week time period following the placement of silk ligatures and initiation of a soft diet. Three animals received intrapapillary injections of soluble receptors (blockers), capable of blocking the biologic activity for both IL-1 and TNF-alpha, and 3 animals received vehicle (control) injections. Injections were given 3X per week over the course of the study. Clinical assessments included a gingival index and quantification of gingival crevicular fluid (GCF) levels. Collected GCF samples were then used in the biochemical assessment of pyridinoline (PYD) and bone alkaline phosphatase (BAP). Radiographic assessment was made using computer-assisted subtraction radiography to measure both bone density (CADIA) values and linear changes in crestal bone height. RESULTS: Significant (p<0.01) changes using both radiographic measures occurred between 2 and 4 weeks following initiation of disease in this model. The use of the blockers significantly (p<0.01) reduced the levels of radiographic bone loss by approximately 50% over that found in the control sites. Both biochemical markers showed the greatest increase during the first two weeks of the study with PYD levels increased 35-fold over baseline levels after 1 week. This difference in response was significantly (p<0.05) greater than the levels found in the non-ligated teeth or in the ligated teeth receiving blockers injections. BAP levels showed significant increases in ligated teeth compared to non-ligated teeth, but failed to show any significant differences between animals treated with vehicle and those treated with IL-1/TNF antagonists. In contrast to these radiographic and biochemical effects, there were no significant differences detected between animals treated with antagonists and the control group for any of the clinical measures. CONCLUSIONS: The results of this study demonstrate that both subtraction radiography and PYD crevicular fluid levels can detect relative differences in periodontal disease progression, while BAP crevicular fluid levels cannot. PMID- 11895542 TI - Bone reactions at implants subjected to experimental peri-implantitis and static load. A study in the dog. AB - AIM: The aim of the present experiment was to study peri-implant tissue reactions to lateral static load at implants subjected to experimental mucositis or peri implantitis. MATERIAL AND METHODS: 5 beagle dogs were used. The mandibular premolars were extracted. After 12 weeks, 3 implants were installed in each quadrant of the mandible. In one side, the implants were designed with a SLA surface and in the contralateral side with a turned surface. A plaque control program was initiated. 12 weeks later, the central and posterior implants were connected with an appliance containing an expansion screw. Cotton ligatures were placed around the neck of the anterior and posterior implants in both sides, and the plaque control measures were terminated. Sixteen weeks later the ligatures were removed. After 8 weeks without ligatures, the expansion screws in both sides were activated. Once every 2 week during a 12-week interval, the screws were reactivated. Thus, the model included 3 different experimental sites of each surface group: group M+L (mucositis+load); group P (peri-implantitis); group P+L (peri-implantitis+load). Fluorochrome labels were injected and standardized radiographs obtained. The animals were sacrificed and block biopsies of all implant sites dissected and prepared for histological analysis. RESULTS: It was demonstrated that the lateral static load failed to induce peri-implant bone loss at implants with mucositis and failed to enhance the bone loss at implants with experimental peri-implantitis. The proportion of bone labels and the bone density in the interface zone were significantly higher in group P+L than in group P. CONCLUSION: It is suggested that a lateral static load with controlled forces may not be detrimental to implants exhibiting mucositis or peri-implantitis. PMID- 11895543 TI - Distribution of erm(F) and tet(Q) genes in 4 oral bacterial species and genotypic variation between resistant and susceptible isolates. AB - BACKGROUND: Bacteroides forsythus, Porphyromonas gingivalis and Prevotella intermedia are Gram-negative anaerobic bacteria that are currently considered potential periopathogens. Prevotella nigrescens has recently been separated from P. intermedia and its role in periodontitis is unknown. The erm(F) gene codes for an rRNA methylase, conferring resistance to macrolides, lincosamides and streptogramin B (MLSB), and the tet(Q) gene for a ribosomal protection protein, conferring resistance to tetracycline. The presence of these resistance genes could impair the use of antibiotics for therapy. PURPOSE: The aim of this study was to determine the carriage of erm(F) and tet(Q), and genetic variability of 12 Porphyromonas gingivalis, 10 Prevotella intermedia, 25 Prevotella nigrescens and 17 Bacteroides forsythus isolates from 9 different patient samples. METHODS: We used polymerase chain reaction (PCR) for detecting antibiotic resistance genes, and pulsed-field gel electrophoresis (PFGE) for detecting genetic variability among the isolates. RESULTS: Thirty-one (48%) isolates were resistant to both erythromycin and tetracycline and carried the erm(F) and tet(Q) genes, eight (13%) were tetracycline resistant and carried the tet(Q) gene, 9 (14%) were erythromycin resistant and carried the erm(F) gene, and 12 (19%) isolates did not carry antibiotic resistance genes. PFGE was used to compare isolates from the same patient and isolates from different patient samples digested with XbaI. No association was found between antibiotic resistance gene carriage and PFGE patterns in any species examined. All isolates of the same species from the same patient had highly related or identical PFGE patterns. Isolates of same species from different patients had unique PFGE pattern for each species tested. CONCLUSION: All isolates of the same species from any one patient were genetically related to each other but distinct from isolates from other patients, and 66% of the patients carried antibiotic resistant isolates, which could impair antibiotic therapy. PMID- 11895544 TI - Subgingival microbiota of indigenous Indians of Central America. AB - OBJECTIVE: To define the subgingival microbial profiles of adult subjects from a previously identified rural community of indigenous Indians in Guatemala, Central America. MATERIALS AND METHODS: A full-mouth periodontal examination was performed in 114 adult subjects from 45 families. Plaque samples were collected from both deep and shallow periodontal pockets and checkerboard DNA-DNA hybridization was employed to identify 17 species previously associated with periodontitis or health. RESULTS: Plaque deposits and gingivitis were universal and widespread, and periodontal pocketing > or =5 mm was highly prevalent (84% of subjects). Streptococcus sanguis, Actinomyces naeslundii genospecies 2 and Fusobacterium nucleatum were significantly more prevalent in shallow sites. At the subject level, Actinomyces naeslundii and Peptostreptococcus micros were significantly more prevalent in periodontally-healthy subjects. Actinobacillus actinomycetemcomitans was not detected in any sample. CONCLUSION: There was no association between periodontal disease status and presence of suspected periodontal pathogens. These latter results conflict somewhat with those from treated populations. However, in this population where extensive plaque deposits and gingivitis are universal, the presence of putative pathogens may be more reflective of the local environment. PMID- 11895545 TI - Clinical and microbial evaluation of a histatin-containing mouthrinse in humans with experimental gingivitis: a phase-2 multi-center study. AB - OBJECTIVE: P-113, a 12 amino acid histatin-based peptide, was evaluated in a mouthrinse formulation for safety and efficacy in a phase 2 multi-center clinical study. METHOD: 294 healthy subjects abstained from oral hygiene procedures and self-administered either 0.01% P-113, 0.03% P-113 or placebo mouthrinse formulations twice daily over a 4-week treatment period. During this time, the safety, anti-gingivitis, and anti-plaque effects of P-113 were evaluated. RESULTS: There was a significant reduction in the change from baseline to Day 22 in bleeding on probing in the 0.01% P-113 treatment group of the intent to treat population (p=0.049). Non-significant trends in the reduction of the other parameters were observed in this population (p> or =0.159). A sub-group of subjects which developed significant levels of disease within the four-week timeframe of the study was identified based on baseline gingival index scores > or =0.75. Significant findings were observed for bleeding on probing, gingival index and plaque index within this population (p<0.05). There were no treatment related adverse events, and there were no adverse shifts in supragingival microflora during the study. Significant amounts of the peptide were retained in the oral cavity following rinsing. CONCLUSION: These data suggest that P-113 mouthrinse is safe and reduces the development of gingival bleeding, gingivitis and plaque in the human experimental gingivitis model. PMID- 11895546 TI - Effect of interleukin-1 gene polymorphism in a periodontally healthy Hispanic population treated with mucogingival surgery. AB - OBJECTIVES: A genetic test for susceptibility of periodontal disease has been introduced. A positive test indicates a risk factor for more severe periodontal destruction. The prevalence of genotype positive subjects has been reported around 30%. In a Mexican population, we have found a 26% prevalence of genotype positive individuals. Few studies have reported the response to therapy in these individuals. The purpose of this study was to assess the response to mucogingival surgery in an otherwise periodontally healthy Hispanic population. MATERIALS AND METHODS: 22 subjects (7 male and 15 female) with a mean age of 45 years participated. They were treated 3 years prior for the treatment of Types I and II recession defects using connective tissue grafts. No other active periodontal treatment was required, except for preventive maintenance. A full-mouth clinical evaluation was performed which included assessment of gingival inflammation and measurements of probing pocket depth and clinical attachment levels. Mean values per patient were determined. A finger stick blood sample was collected using specially provided DNA filter paper, let dried, and mailed for processing. RESULTS: Results indicated that 5 out of the 22 subjects were genotype positive. The genotype positive subjects presented the following values: GI 1.13+/-0.17, PPD 2.48+/-0.46, and CAL 3.38+/-0.66. The values for the genotype negative subjects were GI 1.06+/-0.14, PPD 2.38+/-0.31 and CAL 3.11+/-0.53. No statistical significant differences were found when both groups were compared (p>0.05). Furthermore, the treatment of the localized recessions was effective and provided similar amount of coverage in genotype positive and negative subjects. However, more genotype negative subjects showed complete coverage of the recession than genotype positive individuals. CONCLUSIONS: Within the limits of this study it is concluded that (1) periodontal health can be maintained with proper preventive maintenance irrespective of the genotype present, (2) the mean response to mucogingival surgery to cover localized gingival recessions is similar irrespective of the IL-1 periodontal genotype, however, full coverage is achieved more frequently in genotype negative subjects. PMID- 11895547 TI - Primary gingival leiomyosarcoma. A clinicopathological study of 1 case with prolonged survival. AB - BACKGROUND: Leiomyosarcoma is a relatively uncommon mesenchymal tumor that exhibits smooth-muscle differentiation. Only 3 to 10% of leiomyosarcomas arise in the head and neck, the nose and paranasal sinuses, skin and subcutaneous tissue and cervical esophagus being the most common localizations. Most leiomyosarcomas involving the oral tissues primarily affect the maxillary sinus, the maxillary or mandibular bone. A review of the English-language literature since 1908 revealed 30 reported cases of primary leiomyosarcoma of the oral mucosa and soft tissues. MATERIAL AND METHODS: We report on a case of gingival leiomyosarcoma, arising in a 31-year-old female and involving the upper alveolar mucosa. Following the diagnosis of malignant neoplasm on frozen sections and an en-block resection, the tumour was formalin-fixed and paraffin embedded for histological and immunohistochemical examination. RESULTS: Microscopically, the tumor was composed of interlacing fascicles of spindle-shaped cells with elongated, blunt-ended nuclei and eosinophilic cytoplasm, containing PAS-positive granules. Mitoses, both typical and atypical, and scattered necrotic foci were present. Consistent desmin, muscle specific and alpha-smooth muscle-specific, and vimentin immunoreactivity was demonstrated in the tumor cells. The patient is alive and free of disease at a 7-year follow-up. CONCLUSIONS: Intra-oral leiomyosarcomas are exceptionally rare. Accurate diagnosis and treatment is largely based on the careful search of clinical signs indicative of malignancy (e.g., neoplastic bone destruction, wide invasion of adjacent tissues) and intra-operative (frozen sections) examination of the lesion. Though the case reported herein showed an attenuated clinical behavior, prolonged follow-up is mandatory in view of possible tumor relapse. PMID- 11895548 TI - Non-cirrhotic portal hypertension: why is it so common in India? PMID- 11895549 TI - Non-cirrhotic portal fibrosis (idiopathic portal hypertension): experience with 151 patients and a review of the literature. AB - BACKGROUND: Non-cirrhotic portal fibrosis (NCPF), the equivalent of idiopathic portal hypertension in Japan and hepatoportal sclerosis in the United States of America, is a common cause of portal hypertension in India. The clinical features, portographic and histological findings, and management of 151 patients with non-cirrhotic portal fibrosis are presented. METHODS: The disease is diagnosed by the presence of unequivocal evidence of portal hypertension in the definite absence of liver cirrhosis and extrahepatic portal vein obstruction (EHPVO). Retrospective analysis of records of 151 patients with NCPF was analyzed for the clinical presentation, physical findings, laboratory tests, radiological and histological findings, and for the outcome of treatment. RESULTS: The disease is characterized by massive splenomegaly with anemia, preserved liver function and benign prognosis in a majority of patients. Splenoportovenography (SPV) showed massive dilatation of the portal and splenic veins, and the presence of collaterals. Twenty-four (15.9%) patients showed evidence of natural/spontaneous shunts (splenorenal 15, umbilical nine) on SPV; these patients had a lower incidence of variceal bleeding. Liver histology demonstrated maintained lobular architecture, portal fibrosis of variable degree, sclerosis and obliteration of small-sized portal vein radicles, and subcapsular scarring with the collapse of the underlying parenchyma. Piecemeal or hepatocytic necrosis was absent in all histology specimens. Three patients showed nodular transformation along with abnormal liver functions, and may represent late manifestation of NCPF where features are similar to those seen in patients with incomplete septal cirrhosis. In the initial part of the study, surgery (side-to-side lieno-renal shunt) was the preferred modality of treatment, however, endoscopic sclerotherapy or variceal ligation has now become the preferred first line of management of variceal bleeding. CONCLUSIONS: The epidemiological and clinical features of NCPF have more similarity to IPH than has previously been documented. The development of spontaneous shunts tends to protect these patients from variceal bleeding. PMID- 11895551 TI - Characteristic endoscopic and magnified endoscopic findings in the normal stomach without Helicobacter pylori infection. AB - BACKGROUND AND AIMS: The aim of this study was to clarify the endoscopic features of the Helicobacter pylori (H. pylori)-free stomach by examining the arrangement of minute points visible on the corpus. Since these points were clarified by magnifying endoscopy as collecting venules, this finding was termed 'regular arrangement of collecting venules (RAC)'. The findings from more endoscopic studies are presented and the differences between magnified views of the normal and H. pylori-infected corpus and antrum are described in particular. METHODS: The study group consisted of 557 patients who were subjected to endoscopy and checked for H. pylori. The RAC in each patient was assessed. Magnifying endoscopy in 301 patients was used to examine the corpus and in 94 patients to examine the antrum. RESULTS: One hundred and fifty-eight patients had normal stomachs without H. pylori. We diagnosed 389 patients with H. pylori gastritis. In 10 patients H. pylori was not detected, but inflammation was present. Of the 158 patients with H. pylori-negative normal stomachs, 151 had RAC. As a determinant of the normal stomach without H. pylori infection, the presence of RAC had 93.8% sensitivity and 96.2% specificity. All 30 patients with H. pylori-negative normal stomachs had a well-defined ridge pattern (wDRP) on the antrum as observed under magnifying endoscopy. As a determinant of the normal stomach without H. pylori infection, wDRP had a specificity of 100%, but a sensitivity of only 54.5%. CONCLUSIONS: The presence of RAC is characteristic of a normal stomach without H. pylori. Magnified views of the normal antrum were different from that of the normal corpus. PMID- 11895550 TI - Fas (Apo-1/CD95) and Fas ligand interaction between gastric cancer cells and immune cells. AB - BACKGROUND AND AIMS: It has been proposed that the expression of Fas ligand (Fas L) in tumors may play an important role in immune escape. This study was undertaken to test a 'counterattack' theory as a mechanism of immune escape in gastric carcinoma. METHODS: Expression of Fas and Fas L was examined in the human gastric cancer cell lines using reverse transcription-polymerase chain reaction. Cytotoxicity was determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. Apoptosis of target Jurkat cells was examined after coculture with the effector gastric cancer cells in vitro. Immunohistochemical staining was performed for the detection of Fas and FasL in tumor-infiltrating lymphocytes (TIL) and gastric cancer cells in vivo. Apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) method in vitro and in vivo. RESULTS: Fas and FasL mRNA were found to be differentially expressed in gastric cancer cell lines. The coculture experiment showed that apoptosis of Jurkat was induced by a FasL-overexpressing effector gastric cell SNU-484. In a Fas-expressing gastric cell SNU-638, Fas expression was upregulated by the treatment of gamma-interferon in a time- and concentration dependent manner. SNU-638 treated with gamma-interferon was more sensitive to anti-Fas antibody-mediated cytotoxicity than was the control cell line, suggesting an increase of functional Fas in gastric cancer cells. The expression of FasL in gastric cancer cells and of Fas in apoptotic TIL was also detected in vivo. CONCLUSION: The data indicate that the FasL expression of gastric cancer cells supports a 'counterattack theory' in gastric cancer cells and that the upregulation of Fas by IFN-gamma in SNU-638 may accelerate the apoptosis pathway through the Fas and FasL interaction between gastric cancer cells and immune cells. This result is supported by the expression of FasL in gastric cancer cells and apoptotic TIL in vivo. It is implicated that the different biological behaviors of gastric cancer cells could be at least in part explained by Fas and FasL interaction with immune cells. PMID- 11895552 TI - Mediation of transcatheter arterial chemoembolization induced gastric slow-wave dysrhythmia by endogenous prostaglandin. AB - BACKGROUND AND AIMS: In recent years, gastric slow-wave dysrhythmias induced by transcatheter arterial chemoembolization (TACE) have been observed. Enhanced endogenous prostaglandin may be a possible mechanism for the myoelectrical changes. The aim of this study was to evaluate whether the gastric slow-wave dysrhythmias induced by TACE may be mediated by ketoprofen, a prostaglandin synthesis inhibitor. METHODS: Twenty-three patients with hepatocellular carcinoma (HCC) admitted for TACE were enrolled. A follow-up TACE was scheduled to take place 2 months later. During the next admission for TACE, 50 mg of ketoprofen was given intramuscularly 12 h for 3 days, beginning 48 h before TACE, as premedication. Cutaneous electrogastrography (EGG) was performed before and within 24 h after TACE. RESULTS: The results showed that the change in the fasting EGG parameters after TACE without premedication was not statistically significant. However, the postprandial EGG parameters, including the dominant frequency (DF); the percentages of DF in the normal, bradygastric and tachygastric range; along with the dominant frequency instability coefficient, deteriorated significantly after the procedure (P < 0.01). After the follow-up TACE with ketoprofen premedication, neither the fasting nor postprandial EGG parameters in the control group changed significantly. CONCLUSIONS: Gastric slow wave dysrhythmias induced by TACE may be mediated by ketoprofen, a prostaglandin synthesis inhibitor, in HCC patients. However, the improvement in the gastric myoelectrical activity does not eliminate the degree of nausea/vomiting after TACE. PMID- 11895553 TI - Recurrences of hepatocellular carcinoma following initial remission by transcatheter arterial chemoembolization. AB - BACKGROUND AND AIMS: The aim of this study was: (i) to define the characteristics of hepatocellular carcinoma (HCC) associated with recurrences following initial remission by transcatheter arterial chemoembolization (TACE); (ii) to evaluate the patterns of recurrences; and (iii) find a better surveillance method of detecting recurrent HCC. METHODS: Out of 230 consecutive HCC patients who underwent TACE, 77 with initial remission were followed prospectively for at least 12 months. We compared the recurrence rates according to the characteristics of the tumors and analyzed the locations of the recurrent HCC. The diagnostic efficacies of CT scans with serum AFP, angiography and Lipiodol CT scan in detecting recurrent HCC were also evaluated. RESULTS: Recurrent HCC was detected in 40 patients during a median period of 27 months. The recurrence rate of multinodular HCC was higher than the single nodular type. All six patients with portal vein thrombosis recurred. Even though 45% of recurrences were adjacent to original tumors, 63% were separated from them (8% at both). Hepatocellular carcinoma with heterogeneous Lipiodol uptake tended to recur at the site adjacent to the original tumors more frequently than HCC with homogeneous Lipiodol uptake. Only 18 of 40 recurrent HCC were initially detected by serum alpha-fetoprotein (AFP) and CT scans: 19 by angiography and three only by Lipiodol CT scan. CONCLUSION: Our data indicated that HCC of the multinodular type and with portal vein thrombosis recur more frequently following initial remission by TACE. It is also suggested that regular angiography in addition to serum AFP and CT scan may be valuable in detecting recurrent HCC. Other treatment modalities may need to be combined to ablate tumors completely and to therefore reduce recurrences, especially in HCC with heterogeneous Lipiodol uptake. PMID- 11895554 TI - Role of the spleen in liver fibrosis in rats may be mediated by transforming growth factor beta-1. AB - BACKGROUND: The effect of the spleen on the cirrhotic liver is unknown. Transforming growth factor-beta 1 (TGF-beta 1), which plays a crucial role in the matrix production during liver fibrosis, is an inhibitory factor regarding the regeneration of hepatocytes. In this study, we investigated the TGF-beta 1 production in the spleen of cirrhotic rats and the effects of a splenectomy on the healing process from liver fibrosis. METHODS: Thirty-six Wistar male rats were used. Thioacetamide (TAA) was administered intraperitoneally for 24 weeks. The rats underwent either a sham operation (TAA + Sham) or a splenectomy (TAA + SPL). The improvements in liver fibrosis and liver regeneration were investigated 10, 30 and 60 days after the operations in each group. The effect of a splenectomy on the plasma concentration of TGF-beta 1 in the portal vein was investigated by ELISA. The TGF-beta 1 expressions in the spleen were measured using immunohistochemical staining and the degree of such expression was measured using RT-PCR. The activity of TGF-beta 1 in the portal vein of TAA + Sham and TAA + SPL was assessed by the inhibiting effect of rat parenchymal hepatocyte proliferation in primary culture. RESULTS: Liver regeneration (PCNA-labeling index) in the TAA + SPL rats was stimulated more at 10 and 30 days after the operation (P < 0.05) than in the TAA + Sham rats, and the improvement of liver fibrosis (fibrosis rate) in the TAA + SPL rats was higher at 60 days (P < 0.05) than in the TAA + Sham rats. The plasma concentration of TGF-beta1 of the portal vein in TAA + SPL rats was significantly lower than in the TAA + Sham rats for each period. Immunohistochemically, TGF-beta1-positive stained cells were recognized in the spleen macrophages in the red pulp of cirrhotic rats. The plasma of the TAA + Sham rats at 10 and 30 days after the operation was significantly stronger than that of the TAA + SPL rats in inhibiting the proliferation of rat hepatocytes of primary culture. Inhibitory effects were then dose-dependently neutralized by monoclonal TGF-beta 1 antibody. CONCLUSION: Spleen-derived TGF-beta 1 may thus play an inhibitory role in the healing of liver cirrhosis by inhibiting the regeneration of the damaged liver. PMID- 11895555 TI - Hepatic covalent adduct formation with zomepirac in the CD26-deficient mouse. AB - BACKGROUND AND AIMS: Zomepirac (ZP), a non-steroidal anti-inflammatory drug (NSAID), has been reported to cause immune-mediated liver injury. In vivo, ZP is metabolized to a chemically reactive acyl glucuronide conjugate (ZAG) which can undergo covalent adduct formation with proteins. Such acyl glucuronide-derived drug-protein adducts may be important in the development of immune and toxic responses caused by NSAID. We have shown using immunoabsorptions that the 110 kDa CD26 (dipeptidyl peptidase IV) is one of the hepatic target proteins for covalent modification by ZAG. In the present study, a CD26-deficient mouse strain was used to examine protein targets for covalent modification by ZP/metabolites in the liver. METHODS AND RESULTS: The CD26-deficient phenotype was confirmed by immunohistochemistry, flow cytometry analysis, RT-PCR, enzyme assay and immunoblotting. Moreover, by using monoclonal antibody immunoblots, CD26 was not detected in the livers of ZP-treated CD26-deficient mice. Immunoblots using a polyclonal antiserum to ZP on liver from ZP-treated mice showed three major sizes of protein bands, in the 70, 110 and 140 kDa regions. Most, but not all, of the anti-ZP immunoreactivity in the 110 kDa region was absent from ZP-treated CD26 deficient mice. CONCLUSION: These data definitively showed that CD26 was a component of ZP-modified proteins in vivo. In addition, the data suggested that at least one other protein of approximately 110 kDa was modified by covalent adduct formation with ZAG. PMID- 11895556 TI - Therapeutic vaccination in chronic hepatitis B. AB - AIMS: The aim was to test the efficacy of a pre-S2-containing vaccine (Genhevac B) in chronic hepatitis B (CHB). Twenty-five naive patients (22 male, three female; median age 35; range: 6-69 years) with CHB were recruited. The inclusion criteria were: hepatitis B e antigen (HBeAg) positive or HBV-DNA detectable with liquid hybridization; alanine aminotransferase (ALT) is at least 1.5-fold the upper normal limit and histological evidence of chronic hepatitis. METHODS: In the first period, all patients received monthly injections of 20, 40 and 60 microg of the vaccine. One month after the last injection, patients who still had HBV-DNA were divided into two randomly assigned groups. While the patients in the first group and the patients who lost HBV-DNA in the first period continued to receive monthly injections of 20 microg vaccine for a further 6 months, the patients in the second group received 9 MU interferon alpha-2b (Roferon-A), three times per week using the same method as for the first group. Patients were followed up after 12 months without treatment. Response was defined as the loss of HBV-DNA and normalization of ALT. RESULTS: Six of the 25 patients lost HBV-DNA after 3 months. Nine of the remainder were randomly placed in the first group (vaccine-only) and 10 were placed in the second group (vaccine + interferon). End of-treatment response was achieved, overall, 8/15 from the vaccine group and 6/10 from the combination. One patient from each group relapsed during the follow up. Overall, the sustained response (SR) rate was 46% (7/15) in the vaccine group, and 50% (5/10) in the combination group. Histological improvement was achieved in 6/7 SR with vaccine-only and all five with combination treatment, while 1/8 of failures of vaccine and 2/5 of failures of combination improved. CONCLUSIONS: It was concluded that Genhevac-B decreases serum HBV-DNA levels in the majority of patients with CHB and sustained clearance was achieved in some patients. Combination of interferon-alpha with Genhevac-B is effective for the vaccine failures and may increase sustained response compared to interferon-alpha alone. However, the mechanism of action is yet to be explained. PMID- 11895557 TI - Effects of splenectomy on liver volume and prognosis of cirrhosis in patients with esophageal varices. AB - BACKGROUND: Several previous studies have shown that hepatic regeneration after partial hepatic resection accelerates over time once a splenectomy has been performed. This was a retrospective study investigating whether a splenectomy has some beneficial effects for cirrhotic patients with esophageal varices. METHODS: Ninety-three patients underwent either esophageal transection, including splenectomy (splenectomy group), or endoscopic injection sclerotherapy (controls) for esophageal varices. No patient had hepatocellular carcinoma and the grades of their hepatic function were from mild to moderate. The changes in hepatic and splenic functions and liver volume were evaluated, as well as the probability of survival. RESULTS AND CONCLUSIONS: Both plasma white blood cell and platelet counts significantly increased in the splenectomy group compared to the controls (P < 0.05). The proportion of liver volume 1 year after the treatments compared to the volume before the treatments (which was 100%) was 96.4% in splenectomy group and 94.4% in controls. No patient had serious complications, such as severe infection caused by the splenectomy. The two groups showed no statistically significant differences in survival rates throughout this study. Although hypersplenism significantly was improved by splenectomy, no difference in changes in liver volume nor survival probability between the two groups was found. Further studies, such as those with a large number of patients, long-term volumetric analysis, or histopathological examination, are needed to clarify fully the effects of splenectomy on cirrhotic patients. PMID- 11895558 TI - Systemic administration of liposome-encapsulated OK-432 prolongs the survival of rats with hepatocellular carcinoma through the induction of IFN-gamma-producing hepatic lymphocytes. AB - BACKGROUND: OK-432 is known to increase the host antitumor response. We previously reported that systemic administration of OK-432 (OK-Lipo) specifically induced hepatic lymphocytes in mice. Here we aimed to investigate the antitumor effect of OK-Lipo on hepatocellular carcinomas (HCC) in experimental rats. METHODS: Diethylnitrosamine was administered for 12 weeks to all rats (n = 36). Rats were divided into three groups of 12 rats each. One group was injected with OK-Lipo from week 5 (OK-5w group) and another from week 9 (OK-9w group). A control group was injected with saline from week 5 (Non-OK group). At week 13, five rats from each group were used for histological analysis and immunofluorescence assays (surface phenotypic and intracellular cytokine analysis of the mononuclear cells in the liver, spleen and peripheral blood). The remaining rats were observed for the remainder of their survival period. RESULTS: The mean survival times of Non-OK, OK-5w, and OK-9w groups differed significantly (98.0 +/- 5.3 days, 116.0 +/- 5.8 days, and 106.0 +/- 5.4 days, respectively, P < 0.01). Histological examination revealed many apoptotic tumor cells, infiltration of lymphocytes and macrophages in the OK-5w group. The two-color immunofluorescence assay showed that the proportion of natural killer (NK) cells and IFN-gamma-producing cells in the liver were significantly higher in the OK-5w group. CONCLUSIONS: These findings showed that systemic administration of OK-Lipo contributed to prolonging the survival of rats with HCC. OK-Lipo induced NK cells and IFN-gamma-producing cells specifically in the liver and these cells seemed to reduce hepatocarcinogenesis and tumor growth. PMID- 11895559 TI - Case-control study of calcification of the hepatic artery in chronic hemodialysis patients: comparison with the abdominal aorta and splenic artery. AB - BACKGROUND AND AIMS: Studies of the hepatic artery are scarce. We have observed that hepatic artery calcification is very uncommon in patients with hyperparathyroidism that expedites calcification. METHODS: Plain abdominal CT was studied in 221 patients on chronic hemodialysis. Control consisted of 442 sex- and age-matched patients with other diseases. Calcification was graded as a percentage of the entire wall circumference for the aorta, and as a percentage of the entire length of the hepatic and splenic arteries from the celiac trunk to the hilum of each organ. RESULTS: Aortic calcification was seen in 79.2% of male dialysis patients, 22.1% of controls, 74.1% of female dialysis patients and 17.3% of controls (P < 0.0001). Hepatic artery calcification was seen in only 13 dialysis patients. The degree of calcification of the abdominal aorta was correlated with the length of hemodialysis period (P = 0.008), but not with serum calcium, serum phosphate or their product. Although serum parathormone levels were not correlated with calcification, seven of eight dialysis patients with hepatic artery calcification had very high parathormone levels. CONCLUSIONS: The hepatic artery is far less frequently calcified than are the abdominal aorta and splenic artery. This may be a teleologic phenomenon of the liver. PMID- 11895560 TI - The additive effect of opioids and nitric oxide in increasing pentylenetetrazole induced seizure threshold in cholestatic mice. AB - BACKGROUND: Accumulation of endogenous opioids and overproduction of nitric oxide has been reported in cholestatic mice. It is well known that endogenous opioids and nitric oxide alter the susceptibility of experimental animals to different models of seizure. METHODS: The alterations in clonic seizure thresholds, induced by pentylenetetrazole from 1 to 6 days after bile duct ligation, were evaluated in mice. Whether the pretreatment of cholestatic mice with different doses of opioid receptor antagonist naltrexone, or nitric oxide inhibitor N(omega)-nitro-L arginine methyl ester would have changed the clonic seizure threshold was also examined. RESULTS: While in sham-operated mice the clonic seizure threshold was similar to that of the thresholds in unoperated controls, a time-dependent increase in the threshold was observed in cholestatic mice, reaching a peak on day 3 after bile duct ligation and declining partially after the 4th day. Chronic pretreatment with naltrexone (2, 5 and 10 mg/kg, i.p.) reversed the increased threshold in cholestatic mice on day 3 after operation in a dose-dependent manner with the highest doses used restoring the threshold to that of the control animals. A similar reversal of the increased threshold was observed after acute (1, 3 and 10 mg/kg, i.p.) or chronic (10 mg/kg, i.p. for 4 days) pretreatment with N(omega)-nitro-L-arginine methyl ester. Moreover, concurrent administration of doses of N(omega)-nitro-L-arginine methyl ester and naltrexone that each separately induced a partial reversal of increased seizure threshold in cholestasis caused a complete restoring of the threshold to the control level. CONCLUSIONS: Based on these findings, both opioid receptors and nitric oxide may be involved in the dramatic increase in pentylenetetrazole-induced seizure threshold in cholestasis. PMID- 11895561 TI - Gastrointestinal: Clostridium difficile colitis. PMID- 11895562 TI - Hepatobiliary and pancreatic: hepatocellular carcinoma treated with transcatheter arterial chemoembolization. PMID- 11895563 TI - A case of Behcet's disease accompanied by colitis with longitudinal ulcers and granuloma. AB - A 37-year-old female presenting with oral and genital ulcers and erythema nodosum on both arms was diagnosed as having Behcet's disease. The symptoms resolved spontaneously. However, she was admitted to our hospital (Keio University Hospital) several months later because of fever, aphthous ulcers of the oral cavity, lower abdominal pain and frequent diarrhea. A colonoscopic examination revealed multiple ulcers including longitudinal ulcers in the ascending and transverse colon, and histological examination of biopsied specimens demonstrated non-caseating epithelioid granuloma. Treatment with prednisolone and 5 aminosalicylic acid was started, and the patient responded well clinically. One month later, a repeated colonoscopy confirmed that the lesions including longitudinal ulcers had disappeared. In this report, we describe our experience of this rare case of Behcet's disease concomitant with colonic longitudinal ulcers and epithelioid granuloma, and discuss the difficulties in making a differential diagnosis, primarily with regard to Crohn's disease. PMID- 11895565 TI - Gastro-colic fistula complicating primary (Al) amyloidosis. PMID- 11895568 TI - The effects of isatin (indole-2, 3-dione) on pituitary adenylate cyclase activating polypeptide-induced hyperthermia in rats. AB - BACKGROUND: Previous studies have demonstrated that centrally administered natriuretic peptides and pituitary adenylate cyclase-activating polypeptide-38 (PACAP-38) have hyperthermic properties. Isatin (indole-2, 3-dione) is an endogenous indole that has previously been found to inhibit hyperthermic effects of natriuretic peptides. In this study the aim was to investigate the effects of isatin on thermoregulatory actions of PACAP-38, in rats. RESULTS: One microg intracerebroventricular (icv.) injection of PACAP-38 had hyperthermic effect in male, Wistar rats, with an onset of the effect at 2 h and a decline by the 6th h after administration. Intraperitoneal (ip.) injection of different doses of isatin (25-50 mg/kg) significantly decreased the hyperthermic effect of 1 microg PACAP-38 (icv.), whereas 12.5 mg/kg isatin (ip.) had no inhibiting effect. Isatin alone did not modify the body temperature of the animals. CONCLUSION: The mechanisms that participate in the mediation of the PACAP-38-induced hyperthermia may be modified by isatin. The capability of isatin to antagonize the hyperthermia induced by all members of the natriuretic peptide family and by PACAP-38 makes it unlikely to be acting directly on receptors for natriuretic peptides or on those for PACAP in these hyperthermic processes. PMID- 11895567 TI - Universal sequence map (USM) of arbitrary discrete sequences. AB - BACKGROUND: For over a decade the idea of representing biological sequences in a continuous coordinate space has maintained its appeal but not been fully realized. The basic idea is that any sequence of symbols may define trajectories in the continuous space conserving all its statistical properties. Ideally, such a representation would allow scale independent sequence analysis--without the context of fixed memory length. A simple example would consist on being able to infer the homology between two sequences solely by comparing the coordinates of any two homologous units. RESULTS: We have successfully identified such an iterative function for bijective mapping psi of discrete sequences into objects of continuous state space that enable scale-independent sequence analysis. The technique, named Universal Sequence Mapping (USM), is applicable to sequences with an arbitrary length and arbitrary number of unique units and generates a representation where map distance estimates sequence similarity. The novel USM procedure is based on earlier work by these and other authors on the properties of Chaos Game Representation (CGR). The latter enables the representation of 4 unit type sequences (like DNA) as an order free Markov chain transition table. The properties of USM are illustrated with test data and can be verified for other data by using the accompanying web-based tool:http://bioinformatics.musc.edu/~jonas/usm/. CONCLUSIONS: USM is shown to enable a statistical mechanics approach to sequence analysis. The scale independent representation frees sequence analysis from the need to assume a memory length in the investigation of syntactic rules. PMID- 11895569 TI - A comparison of the illness beliefs of people with angina and their peers: a questionnaire study. AB - BACKGROUND: What people believe about their illness may affect how they cope with it. It has been suggested that such beliefs stem from those commonly held within society. This study compared the beliefs held by people with angina, regarding causation and coping in angina, with the beliefs of their friends who do not suffer from angina. METHODS: Postal survey using the York Angina Beliefs Questionnaire (version 1), which elicits stress attributions and misconceived beliefs about causation and coping. This was administered to 164 people with angina and their non-cohabiting friends matched for age and sex. 132 people with angina and 94 friends completed the questionnaire. RESULTS: Peers are more likely than people with angina to believe that angina is caused by a worn out heart (p < 0.01), angina is a small heart attack (p = 0.02), and that it causes permanent damage to the heart (p < 0.001). Peers were also more likely to believe that people with angina should take life easy (p < 0.01) and avoid exercise (p = 0.04) and excitement (p < 0.01). CONCLUSIONS: The beliefs of the peer group about causation and coping in angina run counter to professional advice. Over time this may contribute to a reduction in patient concordance with risk factor reduction, and may help to create cardiac invalids. PMID- 11895570 TI - Prenatal diagnosis of Neu-Laxova syndrome: a case report. AB - BACKGROUND: Neu-Laxova syndrome is a rare congenital abnormality involving multiple systems. We report a case of Neu-Laxova syndrome (NLS) diagnosed prenatally by ultrasound examination. CASE PRESENTATION: A 29-year-old gravida 3, para 2 woman was first seen in our antenatal clinic at 38 weeks' pregnancy. Except for the consanguinity and two previous abnormal stillborn babies her medical history was unremarkable. On ultrasound examination microcephaly, flat forehead, micrognathia, intrauterine growth restriction, generalized edema of the skin, hypoplastic chest, excessive soft tissue deposition of hands and feet, joint contractures and a penis without scrotal sacs were detected. She delivered a 2000 g male fetus. He died five minutes after delivery. Postmortem examination confirmed the diagnosis of Neu-Laxova syndrome. CONCLUSION: Because of the autosomal recessive inheritance of Neu-Laxova syndrome genetic counseling and early-serial ultrasound examination should be performed at risk families. Early diagnosis of the disease may offer termination of the pregnancy as an option. PMID- 11895573 TI - The ten most commonly asked questions about management of congenital heart disease in adults. PMID- 11895574 TI - The athlete's heart: remodeling, electrocardiogram and preparticipation screening. AB - Highly trained athletes show morphologic cardiac changes (ie, athlete's heart) that are the consequence of several determinants, including type of sport, gender, and, possibly, inherited genetic factors. The extent of physiologic cardiac remodeling may occasionally be substantial in highly trained athletes and may raise a differential diagnosis with structural cardiac disease, such as cardiomyopathies. In addition, athletes demonstrate a spectrum of alterations in the 12-lead electrocardiogram (ECG) pattern, including marked increase in precordial R-wave or S-wave voltages, ST segment or T-wave changes, and deep Q waves suggestive of left ventricular hypertrophy, that may raise the possibility of pathologic heart condition, but have also been viewed as a consequence of the cardiac morphologic remodeling induced by athletic conditioning. To evaluate the clinical significance of these abnormal ECGs, the authors compared ECG patterns to cardiac morphology and function (assessed by two-dimensional echocardiography in individual athlete) in a large population of 1005 elite athletes engaged in a variety of sporting disciplines. Forty percent of the athletes had abnormal ECGs, and a subgroup of about 15% showed distinctly abnormal and often bizarre patterns highly suggestive of cardiomyopathies, such as hypertrophic cardiomyopathy, in the absence of pathologic cardiac changes. Such alterations are likely the consequence of athletic conditioning itself and represent another potential component of athlete's heart syndrome. However, such false-positive ECGs represent a potential limitation to the efficacy of routine ECG testing in the preparticipation cardiovascular screening of large athletic populations. PMID- 11895571 TI - Morphological, histochemical, and interstitial pressure changes in the tibialis anterior muscle before and after aortofemoral bypass in patients with peripheral arterial occlusive disease. AB - BACKGROUND: Morphological and electrophysiological studies of ischemic muscles in peripheral arterial disease disclosed evidence of denervation and fibre atrophy. The purpose of the present study is to describe morphological changes in ischemic muscles before and after reperfusion surgery in patients with peripheral occlusive arterial disease, and to provide an insight into the effect of reperfusion on the histochemistry of the reperfused muscle. METHODS: Muscle biopsies were obtained from the tibialis anterior of 9 patients with chronic peripheral arterial occlusive disease of the lower extremities, before and after aortofemoral bypass, in order to evaluate the extent and type of muscle fibre changes during ischemia and after revascularization. Fibre type content and muscle fibre areas were quantified using standard histological and histochemical methods and morphometric analysis. Each patient underwent concentric needle electromyography, nerve conduction velocity studies, and interstitial pressure measurements. RESULTS: Preoperatively all patients showed muscle fibre atrophy of both types, type II fibre area being more affected. The mean fibre cross sectional area of type I was 3,745 microm2 and of type II 4,654 microm2. Fibre type grouping, great variation in fibre size and angular fibres were indicative of chronic dennervation-reinnervation, in the absence of any clinical evidence of a neuropathic process. Seven days after the reperfusion the areas of both fibre types were even more reduced, being 3,086 microm2 for type I and 4,009 microm2 for type II, the proportion of type I fibres, and the interstitial pressure of tibialis anterior were increased. CONCLUSIONS: The findings suggest that chronic ischemia of the leg muscles causes compensatory histochemical changes in muscle fibres resulting from muscle hypoxia, and chronic dennervation-reinnervation changes, resulting possibly from ischemic neuropathy. Reperfusion seems to bring the oxidative capacity of the previously ischemic muscle closer to normal. PMID- 11895575 TI - The case for early statin therapy in acute coronary syndromes. AB - Three large secondary prevention studies have shown that, in patients with a history of cardiovascular disease, statin treatment reduces the risk of further events and lowers overall mortality. In these studies, total mortality was reduced by as much as 30% in high-risk groups and 22% in average-risk groups. However, these studies did not include patients immediately after the coronary event. There are many benefits to early intervention with statin therapy in patients with acute coronary syndromes, including reduction of the risk of a subsequent event, which is highest immediately after the index event. Early treatment may reduce this likelihood in the first months after a coronary event by stabilizing atherosclerotic plaques and improving endothelial function in addition to lowering low-density lipoprotein cholesterol levels. This article reviews the case for early statin therapy in patients with a history of coronary heart disease. Results of clinical studies have now shown early statin therapy to be safe and cost-effective in reducing in-hospital and 6-month mortality. PMID- 11895576 TI - Aldosterone as a mediator in cardiovascular injury. AB - The renin-angiotensin-aldosterone system plays a central role in the development of hypertension and the progression of end-organ damage. Although angiotensin converting enzyme inhibitors and angiotensin II receptor antagonists can initially suppress plasma aldosterone, it is now well established that aldosterone escape may occur, whereby aldosterone levels return to or exceed baseline levels. The classic effects of aldosterone relate mainly to its action on epithelial cells to regulate water and electrolyte balance. However, blood pressure reduction or fluid loss could not account for the results of the Randomized Aldactone Evaluation Study, which showed that a low dose of spironolactone in addition to conventional therapy could decrease the overall risk of mortality by 30% among patients with severe congestive heart failure. The action of aldosterone at nonepithelial sites in the brain, heart, and vasculature is consistent with the presence of mineralocorticoid receptors in these tissues. Aldosterone has a number of deleterious effects on the cardiovascular system, including myocardial necrosis and fibrosis, vascular stiffening and injury, reduced fibrinolysis, endothelial dysfunction, catecholamine release, and production of cardiac arrhythmias. Several studies have now shown vascular and target-organ protective effects of aldosterone receptor antagonism in the absence of significant blood pressure lowering, consistent with a major role for endogenous mineralocorticoids as mediators of cardiovascular injury. The advent of selective aldosterone receptor antagonists such as eplerenone should prove of great therapeutic value in the prevention of cardiovascular disease and associated end-organ damage. PMID- 11895577 TI - Role of controlled septal infarct in hypertrophic obstructive cardiomyopathy. AB - Hypertrophic obstructive cardiomyopathy is a complex and challenging disease. Medical therapy, surgical therapy, and pacing therapy have been used with some success over the years. Nonsurgical septal reduction therapy, also called alcohol septal ablation, has been used recently as a percutaneous catheter-based intervention to improve left ventricular outflow tract obstruction and thereby improve symptoms. The reported results of this procedure have shown impressive reductions in gradient and improvement in symptoms at relatively low risk. The most common complication of the procedure, development of complete heart block requiring a permanent pacemaker, has improved in recent studies with refinements of the procedural technique. Nonsurgical septal reduction therapy has been shown to improve diastolic function, decrease left ventricular hypertrophy and mass, and cause changes at the cellular and molecular level that improve myocardial function. Reported results at 1 year follow-up continue to show benefit, and long term studies are ongoing. PMID- 11895578 TI - Strategies for the prevention of atherosclerotic progression in women. AB - Atherosclerosis is an inflammatory and fibrotic process that begins early in life in females. The pathogenesis involves the oxidation of the low-density lipoprotein molecule aggravated by smoking, hypertension, lipid abnormalities, and hormonal changes. Risks for progression of atherosclerosis can now be tabulated for the female based on age, cholesterol, smoking, high-density lipoprotein, and systolic blood pressure. During the reproductive years, emphasis should be placed on lifestyle changes, but women at increased risk for diabetes should be aggressively treated with lipid-lowering agents. During the menopausal phase of life, an important consideration is the use of hormone replacement along with lifestyle changes, smoking cessation, blood pressure control, and lipid modification. In the female with established coronary heart disease, standard cardiac medications are indicated along with more aggressive approaches to risk factors and target goals for cholesterol, low-density lipoprotein, and triglycerides. The effect of hormone replacement in established coronary artery disease is uncertain. Therefore, strategies for slowing the progression of atherosclerosis should begin during the reproductive years, with particular emphasis in patients during menopause and in patients with coronary heart disease. PMID- 11895579 TI - Occupational safety communication for hazardous goods: the development of a policy in Israel. AB - Israeli policy governing written occupational safety information for carriage and supply of hazardous goods, and procedures for implementation, are described and evaluated for their potential communicative effectiveness, in view of users' linguistic abilities and the language employed. We also consider whether the addressee should include the end-user and the reading-impaired. The evaluation is set in the context of broader Israeli language policy, and comparison is made with communication policies for hazardous goods adopted by the European Union, the UK, and the USA. PMID- 11895580 TI - Safety attitudes and their relationship to safety training and generalised self efficacy. AB - This paper studies safety attitudes, their relationship with safety training behaviour, and generalised self-efficacy. From a sociotechnical perspective, training programs might be used as a mechanism for enhancing attitudes, especially to improve safety and occupational health. Also, self-efficacy allows to enhance training effectiveness. The aim of this paper is to validate a safety attitude scale and to examine its relationship to safety training behaviour and self-efficacy in organisational settings. With data from 140 employees, results show a conceptually meaningful 3-factor solution. Hierarchical multiple regression analysis shows a main effect of safety training behaviour and levels of self-efficacy on safety attitude. Study limitations and its implications on safety training design are also discussed. PMID- 11895582 TI - The damping of off-central impact for selected industrial safety helmets used in Poland. AB - The paper presents a review of head injury criteria and determines an assessment criterion for test results. Parameters characterizing the protection properties of off-central impacted industrial helmets are chosen and the test stands used in the Central Institute for Labour Protection to test those parameters are presented. The idea of damping 2 selected industrial helmets used in Poland is discussed. The test results of the parameters characterizing the protection properties are compared for damped and not damped helmets. PMID- 11895581 TI - Cultural ergonomics in Ghana, West Africa: a descriptive survey of industry and trade workers' interpretations of safety symbols. AB - Globalization and technology transfer have led to the diffusion of risk communications to users from cultures that were not initially viewed as the target users. This study examined industry and trade workers' overall impressions of symbols used to convey varying degrees of hazardousness. Six symbols, including symbols from the American National Standards Institute (ANSI) Z535 Standard (ANSI, 1998) and the International Organization for Standardization (ISO) 3864:1984 Standard (ISO, 1984) were selected. With the exception of the SKULL symbol, results showed wide discrepancies between users' perceptions of the symbols and their intended meanings. Implications for cross-cultural research on warning components and risk communications are discussed. PMID- 11895583 TI - Physical effects of new devices for bricklayers. AB - Bricklaying is a physically demanding job. Bricklayers frequently flex their trunk to pick up bricks and mortar and position these in a wall. The experienced workload is highest working with bricks at 0 to 50 cm from the floor. In this study the effects of 2 devices that have proven to be feasible in practice are evaluated. The 50 cm raise due to the 2 devices is experienced as comfortable, the estimated lumbar compression force was reduced, and observations indicate likewise. It is discussed that the field experiments have many drawbacks. Nevertheless, based on this study in combination with other literature the improvements could be recommended. PMID- 11895584 TI - Ergonomics in practice: physical workload and heat stress in Thailand. AB - This study consists of assessments of the thermal environment and physiological strain in tasks associated with airport, construction, and metal jobs. The number of male and female participants was 108. Environmental heat stress was evaluated with the WBGT index. Physiological strain was evaluated by the relative cardiovascular load (%CVL) based on the measurements of heart rate. Also the increase of body temperature, weight loss, and perceived discomfort were determinated. At work sites the assessments lasted for 2 to 4 hrs for each participant. The mean physiological strain exceeded the level of 30%CVL. Severe peaks (over 60% CVL) were observed in specific tasks being in agreement with perceived discomfort ratings. The increase of body temperature and weight loss in most cases remained within acceptable limits. For the most strenuous tasks, various ergonomic improvements were developed in consultation with workers and managers. PMID- 11895585 TI - Dust emission and efficiency of local exhaust ventilation during cast iron grinding. AB - A method of determining dust emission and efficiency of its removal by means of local exhaust ventilation from machinery has been described. It complies with Standard No. EN 1093-3:1996 (European Committee for Standardization, 1996) and consists in determining air pollution concentrations in the measurement duct used for air removal from the chamber incorporating devices to be tested. The air volume stream that is pumped is measured at the same time. Test results are presented for dust emission and the efficiency of local exhaust ventilation for cast iron grinding by means of manual power tools and a bench-sander. It has been found that application of local exhaust ventilation contributes to a significant reduction of dust emission with efficiency greater than 90%. PMID- 11895586 TI - Integration of ergonomics into hand tool design: principle and presentation of an example. AB - The development of ergonomic tools responds to health protection needs on the part of workers, especially the work related musculoskeletal disorders of the upper limbs and to the development of ergonomic tools to take into account the needs of the factories. Only an ergonomic design process can enable tool manufacturers to meet these requirements. Three factors are involved: integration of ergonomics into the design process, definition of the different ergonomic stages involved, and finally knowledge of the different factors involved in hand tool design. This document examines these 3 elements in more detail and presents briefly a project of research whose main purpose is to integrate ergonomic criteria into a design process. PMID- 11895587 TI - Methods of assessing efficiency of UV-protecting products. AB - A few aspects of skin protection against ultraviolet (UV) light and methods of assessing efficacy of UV-protecting cream or gels have been discussed. UV protection in cream and gels is not only a strategy of the cosmetic industry but a necessity because of human occupational activity and daily habits. PMID- 11895616 TI - Clinical Trials Report. PMID- 11895615 TI - Clinical Trials Report. PMID- 11895588 TI - The cytotoxicity of some organic solvents on isolated hepatocytes in monolayer culture. AB - The cytotoxic effects of volatile and water-insoluble organic solvents (ethylbenzene, tetrachloroethylene, n-hexane) were tested on isolated hepatocytes in monolayer culture by using the 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction assay. All of the tested compounds inhibited metabolic activity of hepatocytes and this effect depended on the concentration of solvents in the incubatory medium. The presence of fetal calf serum in the medium did not change the cytotoxicity of xenobiotics. IC50 values calculated on the basis of the MTT assay indicated that ethylbenzene was more cytotoxic than tetrachloroethylene and n-hexane. Using hepatocyte monolayer culture and the MTT assay to assess cytotoxicity of organic solvents causes many technical problems. It seems that it cannot be used as a rapid, cheap, and credible method. PMID- 11895617 TI - Clinical Trials Report. PMID- 11895619 TI - T cells and effector mechanisms in the pathogenesis of atopic dermatitis. PMID- 11895618 TI - A genetic disorder of lymphocyte apoptosis involving the fas pathway: the autoimmune lymphoproliferative syndrome. AB - Autoimmune lymphoproliferative syndrome (ALPS) is a recently characterized human disorder that typically presents with lymphocyte accumulation in the first few years of life. This is often associated with the development of autoimmunity, most commonly affecting the hematopoietic system. A key laboratory feature is the marked expansion of double-negative (CD4- and CD8-) T cells that express the alpha/beta T-cell receptor. ALPS is associated with defective Fas-mediated lymphocyte apoptosis, and in most patients, this results from a heterozygous mutation in the TNFRSF6 gene encoding Fas. The clinical features of ALPS reveal the importance of the Fas apoptotic pathway in maintaining lymphocyte homeostasis and protecting against autoimmunity and lymphoid malignancy. PMID- 11895620 TI - T cells in drug allergy. AB - In recent years, increasing evidence has indicated an important role for T cells in various drug-induced diseases. A detailed analysis of patients with various drug allergies revealed the existence of drug-specific T cells in the circulation or in eluate from skin infiltration in bullous, pustular, and maculopapular drug eruptions. The drug-specific T cells use the ab-T cell receptor CD4+ or CD8+ and react with drugs acting as haptens (covalently bound to larger molecules, such as penicillins), but also recognize drugs if they are bound only in a labile way to major histocompatibility complex molecules (noncovalent drug presentation). Functional analysis revealed a predominant IL-5 production by drug-specific CD4+ T cells in maculopapular exanthema (MPE) and bullous skin diseases, while patients with acute generalized exanthematous pustulosis have a peculiar T cell subset secreting high amounts of IL-8. Moreover, in MPE CD4+, perforin+ T cells were found in vitro and in immunohistology that had cytotoxic potential and killed keratinocytes in vitro and in vivo. PMID- 11895621 TI - Allergic adverse reactions to sulfonamides. AB - Antimicrobial sulfonamides were the first antimicrobial agents used effectively to treat infectious diseases. However, because they may cause severe adverse drug reactions (ADRs) and because more effective agents have since been developed, sulfonamides now are used for only a few indications in specific groups, such as AIDS patients. Skin reactions, from benign rash to potentially lethal toxidermias, are the most frequent ADRs to sulfonamides. Other major ADRs include acute liver injury, pulmonary reactions, and blood dyscrasias. Although the mechanisms involved have not been fully elucidated, reactive metabolites appear to play a pivotal role. The hydroxylamine and nitroso metabolites of sulfamethoxazole, the most frequently used sulfonamide today, can bind covalently to proteins because of their chemical reactivity, resulting in the induction of specific adverse immune responses. Therefore, changes in the activity of metabolic and detoxification pathways are associated with a greater risk for developing allergic reactions to sulfonamides. Allergies to sulfonamides, particularly sulfamethoxazole (often used in combination with trimethoprim as co trimoxazole), are more frequent in AIDS patients, but the reason for this increased risk is not fully understood. No valid tools are available to predict which patients have a greater risk for developing allergies to sulfonamides. Diagnosis is essential to avoid a possible evolution toward severe reactions and readministration of the offending drug. In patients who absolutely require further treatment, successful desensitization may be achieved. PMID- 11895622 TI - Management of cutaneous drug reactions. AB - Drugs are potent chemicals that often have effects in the body beyond the desired action. These effects may range from mild and expected side effects to dramatic and life-threatening anaphylaxis. Adverse drug reactions account for between 2% and 6% of hospital admissions and may prevent administration of otherwise effective therapeutic agents. Cutaneous and mucocutaneous eruptions are the most common adverse reactions to oral or parenteral drug therapy, and the spectrum ranges from transitory exanthematous rash to the potentially fatal toxic epidermal necrolysis. Different mechanisms, including both immunologic and nonimmunologic, are responsible for cutaneous adverse drug reaction. The treatment of cutaneous drug eruptions essentially rests on accurate history, a thorough physical examination, discontinuation of the offending drug, and supportive care. The management of a cutaneous drug eruption is very much individualized, based on the clinical setting. This review aims to provide a general approach to the patient with a presumed cutaneous drug reaction. PMID- 11895623 TI - Antiepileptic hypersensitivity syndrome: clinicians beware and be aware. AB - Antiepileptic hypersensitivity syndrome is a serious idiosyncratic, non-dose related adverse reaction reported to occur with phenytoin, phenobarbital, carbamazepine, primidone, and lamotrigine. The reaction usually develops 1 to 12 weeks after initiation of therapy with one of the above agents and is recognized by the classic triad of fever, rash, and internal organ involvement. Immediate discontinuation of the suspected anticonvulsant is essential for good outcome. Patients usually are managed supportively with hydration, antihistamines, H(2) receptor blockers, and topical corticosteroids. In severe cases, the use of systemic corticosteroids may be necessary. The use of intravenous immune globulin should be limited to severe cases where Kawasaki disease or idiopathic thrombocytopenic purpura cannot be ruled out. Education of health care professionals and patients is imperative to improving outcomes and prevention of this reaction in the future. PMID- 11895624 TI - Idiopathic anaphylaxis. AB - Anaphylaxis represents the maximal variant of an immediate-type allergic reaction involving the whole organism with manifestations in different organ systems. IgE mediated mast cell and basophil activation is the major pathomechanism; however, immune complex and pseudo-allergic reactions also may lead to the same symptomatology. The most common elicitors are drugs, additives, occupational substances, animal venoms, aeroallergens, and contact urticariogens but also physical factors (cold, heat, ultraviolet light, exercise). When no eliciting factors can be detected, the term "idiopathic anaphylaxis" is used. The diagnosis of idiopathic anaphylaxis is, therefore, a diagnosis of exclusion and may be made only after careful allergy history taking and diagnosis involving in vitro tests. Possible mechanisms underlying the pathophysiology of idiopathic anaphylaxis include undetected diseases (eg, mastocytosis occulta), concomitant anaphylaxis enhancing medication (b-blockers), secretion of histamine-releasing factor from T lymphocytes, autoantibodies against IgE or IgE receptors, and angiotensin II deficiency. One of the many differential diagnoses of anaphylaxis may have been overlooked. The treatment of idiopathic anaphylaxis follows the rules of antianaphylactic therapy. PMID- 11895625 TI - Hymenoptera (apid and vespid) allergy: update in diagnosis and management. AB - Allergic reactions to Hymenoptera stings range from large local reactions to life threatening anaphylaxis. Over the last 20 years, significant progress has been made using venom extracts in the diagnosis and treatment of Hymenoptera allergy. Despite these advances, there is still room for improvement in increasing the sensitivity of venom allergen skin testing. The venom allergic patient with negative skin tests poses special problems in management. It is important to note their increased risk with a subsequent sting. Guidelines to be used in determining the duration of venom immunotherapy are still evolving. Knowledge of the risks of discontinuing venom immunotherapy and risk factors associated with anaphylaxis with subsequent stings are required to form an individualized approach to treatment. PMID- 11895627 TI - Biotechnology and food allergy. AB - The production of genetically modified foods for an increasingly informed and selective consumer requires the coordinated activities of both the companies developing the transgenic food and regulatory authorities to ensure that these foods are at least as safe as the traditional foods they are supplementing in the diet. Although the size and complexity of the food sector ensures that no single player can control the process from seed production through farming and processing to final products marketed in a retail outlet, checks and balances are in place to ensure that transgenic foods will provide a convenient, wholesome, tasty, safe, affordable food source. Ultimately, it is the responsibility of companies developing the genetically modified food to provide relevant data to regulatory agencies, such as the US Department of Agriculture, Environmental Protection Agency, and Food and Drug Administration, to confirm that the transgenic product is reasonably safe for the consumer, as zero risk from allergen sensitization is nonexistent. PMID- 11895628 TI - How much food is too much? Threshold doses for allergenic foods. AB - This review summarizes recent findings and controversies in the area of threshold doses for allergenic foods. Over the years, there have been many clinical reports that ingestion of small amounts of food can elicit IgE-mediated allergic reactions. In exquisitely allergic individuals, the threshold dose for elicitation of such reactions is often considered to be zero. However, some food allergic patients report that they can tolerate small quantities of allergenic food. Are very low quantities hazardous to food-allergic consumers? How much of the offending food is too much? Why is the concept of a threshold level important? There have been very few studies to date on threshold doses for allergenic foods, and more research is needed in this important area. PMID- 11895629 TI - Eosinophilic esophagitis: an emerging clinicopathologic entity. AB - Eosinophilic esophagitis is a clinicopathologic disease characterized by 1) persistent upper intestinal symptoms despite the patient's use of gastric acid blockade and 2) large numbers of eosinophils in the squamous epithelium of the esophagus. This disease is increasing in frequency and the exact etiologic agent(s) remain elusive. The major importance of identifying eosinophilic esophagitis is that affected patients are receiving fundoplication for persistent symptoms when, in fact, corticosteroids or diet restriction is the treatment of choice. PMID- 11895630 TI - Oral tolerance of food. AB - The gut-associated lymphoid tissue is the specialized arm of the immune system responsible for distinguishing innocuous dietary nutrients, self-proteins, and gut flora from pathogens. Oral tolerance is the active immunologic response to innocuous ingested antigens that leads to a systemic tolerance for that antigen. This process holds great promise for dealing with the host of immune disorders that occur when an inappropriate and destructive immune response is induced. Improved therapies for disorders such as autoimmune disease and allergy are being actively investigated. PMID- 11895631 TI - Occupational reactions to foods. AB - The spectrum of occupational diseases most commonly seen in the food industry includes occupational asthma, rhinitis, conjunctivitis, dermatitis, and hypersensitivity pneumonitis. Occupational asthma represents between 3% and 20% of all asthma cases and is the most common form of occupational lung disease. Occupational skin diseases may represent between 10% and 15% of all occupational diseases, and they have significant economic impact. Hypersensitivity pneumonitis affects the food industry, with farmer's lung representing a common form of the disease. Each of these diseases may have serious and potentially irreversible effects on the health of a farmer, food processor, or food preparer, even after removal of the offending exposure. PMID- 11895633 TI - Mast cell tryptase as a proinflamatory mediator in late-phase asthmatic response. PMID- 11895632 TI - Unproven diagnostic and therapeutic techniques. AB - Over the years, there have been many procedures that either have no diagnostic value for any allergic disease or are inappropriate for the diagnosis and treatment of allergy. These procedures fall into the category of unproven diagnostic and therapeutic techniques for allergy. Unfortunately, there are a very limited number of well-controlled investigations examining these various methods. While these tests may provide a superficial appearance of valid test, they have not been shown by controlled clinical trials to be reliable in the diagnosis and treatment of any allergic diseases. After reviewing the data, it is very clear that there is a need for more well-controlled scientific investigations examining all of these techniques. Until that data becomes available for scientific review and critique, these unproven diagnostic and therapeutic techniques should not be used in the evaluation of patients with suspected allergic disease. PMID- 11895741 TI - [Surgery remains the first weapon against cancer: surgery in French cancer plan. The proposals of the group of French oncologists]. PMID- 11895742 TI - [The Livre Blanc on cancer surgery]. AB - CONTEXT: Among the several therapeutic weapons against cancer, surgical oncology has always been a major, and remains, up today. But in that field, outcomes are heavily dependent on the way the surgery is conducted. OBJECTIVES: The National Federation of Cancer Centers, in accordance with all French cancer specialists, tried to define the adequate conditions allowing the best practices for surgical oncology. This work called Livre blanc, includes several proposals to public authorities. METHOD: Both a specific survey within the French cancer centers and an analysis of public databases have been conducted first. Several experts groups of surgeons and oncologists have been organized, in order to discuss and improve the proposals. RESULTS: Different specificities of surgical oncology are described in the Livre Blanc: The state of medical capacities and it's organization, the impact of the good clinical practice on the outcomes, and the lack of recognition of this speciality. Two main proposals are highlighted: the need of setting up a university diploma better adapted to the main speciality, and the coordination of surgical oncology activity within cancer care networks. PMID- 11895750 TI - [Point of view of the French Society for Digestive Surgery on the Livre Blanc on cancer surgery]. PMID- 11895751 TI - [Thoughts and proposals of the French Association of Urology on the Livre Blanc Paper on cancer surgery]. PMID- 11895752 TI - [Proposals for a sub-speciality of gynecological oncology]. PMID- 11895754 TI - Observations on surgical oncology and the Livre blanc. AB - A high standard of surgical care is the most crucial factor in determining outcome for the great majority of patients with solid malignant tumours. Although acknowledged by most experts this fact is insufficiently recognised by the medical community, by health administrators, by the lay public or by political decision-makers. Difficulty in appreciating the importance of quality in surgery may have come about because of the changing role of the surgeon in cancer care. While remaining the most important instrument in curative cancer treatment, surgery has been joined by other modalities active against malignant disease. PMID- 11895756 TI - p21(CIP1/WAF1/SDI1) hypermethylation: an exciting new lead in ALL biology. PMID- 11895757 TI - The t(14;18) defines a unique subset of diffuse large B-cell lymphoma with a germinal center B-cell gene expression profile. AB - Recently we have identified subgroups of de novo primary diffuse large B-cell lymphoma (DLBCL) based on complementary DNA microarray-generated gene expression profiles. To correlate the gene expression profiles with cytogenetic abnormalities in these DLBCLs, we examined the occurrence of the t(14;18)(q32;q21) in the 2 distinctive subgroups of DLBCL: one with the germinal center B-cell gene expression signature and the other with the activated B cell like gene expression signature. The t(14;18) was detected in 7 of 35 cases (20%). All 7 t(14;18)-positive cases had a germinal center B-cell gene expression profile, representing 35% of the cases in this subgroup, and 6 of these 7 cases had very similar gene expression profiles. The expression of bcl-2 and bcl-6 proteins was not significantly different between the t(14;18)-positive and negative cases, whereas CD10 was detected only in the group with the germinal center B-cell expression profile, and CD10 was most frequently expressed in the t(14;18)-positive cases. This study supports the validity of subdividing DLBCL into 2 major subgroups by gene expression profiling, with the t(14;18) being an important event in the pathogenesis of a subset of DLBCL arising from germinal center B cells. CD10 protein expression is useful in identifying cases of DLBCL with a germinal center B-cell gene expression profile and is often expressed in cases with the t(14;18). PMID- 11895758 TI - 5' CpG island hypermethylation is associated with transcriptional silencing of the p21(CIP1/WAF1/SDI1) gene and confers poor prognosis in acute lymphoblastic leukemia. AB - The p21 is a downstream effector of p53/p73 and belongs to the CIP/KIP family of cyclin-dependent kinase inhibitors (CDKIs). It is, therefore, a potential tumor suppressor gene and probably plays an important role in tumor development. Moreover, reduced expression of p21 has been reported to have prognostic value in several human malignancies. In contrast with other CDKIs, mutational inactivation of p21 is infrequent, but gene inactivation by an alternative mechanism seems to be the general pathway. In this study, we analyzed the methylation status of the p21 promoter region using semiquantitative polymerase chain reaction in 124 patients with acute lymphoblastic leukemia (ALL). We observed p21 hypermethylation in bone marrow cells from 41% (51 of 124) of ALL patients. Hypermethylation within promoter strongly correlated with decreased p21 messenger RNA expression in tumoral cells. Clinical, molecular, and laboratory features and complete remission rate did not differ significantly between hypermethylated and normally methylated patients. Estimated disease-free survival (DFS) and overall survival at 7 and 9 years, respectively, were 59% and 65% for healthy patients and 6% and 8% for hypermethylated patients (P =.00001 and P =.006). Multivariate analysis of potential prognostic factors demonstrated that p21 methylation status was an independent prognostic factor in predicting DFS (P =.0001). Our results indicate that the p21 gene is subject to methylation regulation at the transcription level in ALL and seems to be an important factor in predicting the clinical outcome of these patients. PMID- 11895759 TI - Hydroxyurea corrects the dysregulated L-selectin expression and increased H(2)O(2) production of polymorphonuclear neutrophils from patients with sickle cell anemia. AB - Impaired polymorphonuclear neutrophil (PMN) functions during sickle cell anemia (SCA) may have a pathogenic role in the onset of vasoocclusive events. We used flow cytometry to study, in whole blood, the adhesion molecule expression and respiratory burst of PMNs from children with SCA. Three different clinical groups were studied: (1) patients with no history of vasoocclusive events (n = 15); (2) patients with a history of vasoocclusive events (n = 17); and (3) patients receiving hydroxyurea therapy for severe vasoocclusive events (n = 9). Unstimulated PMNs showed decreased L selectin expression and increased H(2)O(2) production whatever the severity of the disease, reflecting PMN activation. This could contribute to endothelial activation reflected by abnormal plasma levels of soluble adhesion molecules (soluble intercellular adhesion molecule-1, sE selectin, and sL selectin). After stimulation with bacterial N-formyl peptides (N formyl-methionyl-leucyl-phenylalanine [fMLP]), PMNs from untreated patients with a history of vasoocclusive events showed dysregulated L selectin shedding and increased H(2)O(2) production. Furthermore, in these patients, tumor necrosis factor priming followed by fMLP stimulation induced an H(2)O(2) production significantly higher than in the other patient groups and controls. These impairments could immobilize PMNs on the endothelium, thereby inducing reduced blood flow and fostering microvascular occlusion and vascular damage. In contrast, children treated with hydroxyurea showed near-normal basal and poststimulation H(2)O(2) production as well as normal L selectin shedding after stimulation but no change in plasma levels of soluble adhesion molecules. To our knowledge, this is the first report showing major qualitative changes of PMN abnormalities upon hydroxyurea treatment in SCA patients. This strongly suggests that PMNs are a primary target of this drug. PMID- 11895760 TI - Factors affecting duration of survival after onset of blastic transformation of chronic myeloid leukemia. AB - We analyzed factors having an impact on response to treatment and survival in 78 consecutive patients with chronic myeloid leukemia (CML) in blastic transformation (BT) referred to the Hammersmith Hospital from January 1995 to December 2000. BT was defined as the presence of at least 30% blasts in blood or marrow or extramedullary blastic deposits. Immunophenotyping of blasts showed 57 myeloid, 19 lymphoid, and 2 biphenotypic. The median age of the patients was 39.1 years (range, 11.3-73.4 years), with 55 males and 23 females. The median survival for all patients after onset of BT was 8.2 months (95% CI, 6.4-10). Patients in lymphoid BT survived longer than those in myeloid BT (median, 11.2 months versus 6.9 months, P =.052). Initial treatment varied; 41 patients received cytotoxic drugs, 8 underwent allogeneic or autologous transplantation procedures, 21 received STI571 (imatinib mesylate, Gleevec), 1 received radiotherapy, and 7 received no therapy. Of the 25 (32%) patients who achieved a "second chronic phase" with first therapy, 6 of 21 (29%) were treated with STI571 and 19 of 50 (38%) were treated with chemotherapy, transplantation, or radiotherapy. Patients who achieved a second chronic phase survived longer than those who did not (median time from onset of BT 12.0 months versus 6.3 months, P =.0004). In multivariate analysis the finding of more than 50% blast cells in the blood and the presence of cytogenetic progression were independent adverse prognostic variables for survival. We conclude that survival after onset of BT has improved in recent years but is still unsatisfactory. We speculate that the combined use of STI571 with cytotoxic drugs may offer additional benefit. PMID- 11895761 TI - Hepatic sinusoidal obstruction after gemtuzumab ozogamicin (Mylotarg) therapy. AB - Gemtuzumab ozogamicin (Mylotarg) targets leukemia cells expressing the CD33 receptor by means of a monoclonal antibody conjugated to a cytotoxic agent, calicheamicin. Treatment of acute myeloid leukemia (AML) with gemtuzumab ozogamicin may result in liver injury. We reviewed the course of 23 patients who were given gemtuzumab ozogamicin for AML that had relapsed after hematopoietic cell transplantation. Liver toxicity was assessed through physical examination, serum tests, histologic examination, and hepatic venous pressure measurements. Liver injury developed in 11 patients after gemtuzumab ozogamicin administration; it was manifested as weight gain, ascites, and jaundice in 7 patients. Seven patients died with persistent liver dysfunction and either multiorgan failure or sepsis at a median of 40 days after gemtuzumab ozogamicin infusion. Portal pressure measurements were elevated in 2 patients. Results of liver histologic examination in 5 patients showed sinusoidal injury with extensive sinusoidal fibrosis, centrilobular congestion, and hepatocyte necrosis. Six patients experienced AML remission that was sustained for at least 60 days after gemtuzumab ozogamicin infusion. In summary, hepatic sinusoidal liver injury developed after gemtuzumab ozogamicin infusion. Histology showed striking deposition of sinusoidal collagen, suggesting that gemtuzumab ozogamicin targets CD33(+) cells residing in hepatic sinusoids as the mechanism for its hepatic toxicity. PMID- 11895762 TI - Comparative analysis of Ig and TCR gene rearrangements at diagnosis and at relapse of childhood precursor-B-ALL provides improved strategies for selection of stable PCR targets for monitoring of minimal residual disease. AB - Immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements are excellent patient-specific polymerase chain reaction (PCR) targets for detection of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL), but they might be unstable during the disease course. Therefore, we performed detailed molecular studies in 96 childhood precursor-B-ALL at diagnosis and at relapse (n = 91) or at presumably secondary acute myeloid leukemia (n = 5). Clonal Ig and TCR targets for MRD detection were identified in 94 patients, with 71% of these targets being preserved at relapse. The best stability was found for IGK-Kde rearrangements (90%), followed by TCRG (75%), IGH (64%), and incomplete TCRD rearrangements (63%). Combined Southern blot and PCR data for IGH, IGK-Kde, and TCRD genes showed significant differences in stability at relapse between monoclonal and oligoclonal rearrangements: 89% versus 40%, respectively. In 38% of patients all MRD-PCR targets were preserved at relapse, and in 40% most of the targets (> or = 50%) were preserved. In 22% of patients most targets (10 cases) or all targets (10 cases) were lost at relapse. The latter 10 cases included 4 patients with secondary acute myeloid leukemia with germline Ig/TCR genes. In 5 other patients additional analyses proved the clonal relationship between both disease stages. Finally, in 1 patient all Ig/TCR gene rearrangements were completely different between diagnosis and relapse, which is suggestive of secondary ALL. Based on the presented data, we propose stepwise strategies for selection of stable PCR targets for MRD monitoring, which should enable successful detection of relapse in most (95%) of childhood precursor-B-ALL. PMID- 11895763 TI - High hepatitis B virus (HBV) DNA viral load as the most important risk factor for HBV reactivation in patients positive for HBV surface antigen undergoing autologous hematopoietic cell transplantation. AB - The risk factors for hepatitis due to hepatitis B virus (HBV) reactivation in patients positive for hepatitis B surface antigen (HBsAg) treated with autologous hematopoietic cell transplantation (HCT) are unknown. We evaluated 137 consecutive patients (23 positive for HBsAg, 37 positive for hepatitis B surface antibody, and 77 negative for HBV) who underwent HCT. Serial serum ALT were measured before transplant and after transplant at 1 to 4 weekly intervals for the first year and then at 2 to 12 weekly intervals thereafter. Before HCT, basic core promoter (T(1762)/A(1764)) and precore (A(1896)) HBV variants were determined in HBsAg-positive and HBV DNA-positive (by polymerase chain reaction assay) patients by direct sequencing and serum HBV DNA quantitation using the Digene Hybrid Capture II assay. Cox proportional hazards analysis was used to assess the association between pretransplantation HBV virologic and host factors and occurrence of hepatitis due to HBV reactivation. After HCT, hepatitis due to HBV reactivation was more common in HBsAg-positive patients than in HBsAg negative patients (hazard ratio, 33.3; 95% confidence interval [CI], 7.35-142.86; P <.0001). HBsAg-positive patients with detectable serum HBV DNA before HCT (on Digene assay) had a significantly higher risk of hepatitis due to HBV reactivation than HBsAg-positive patients with no detectable serum HBV DNA (adjusted hazard ratio, 9.35; 95% CI, 1.65-52.6; P =.012). Thus, we found that hepatitis due to HBV reactivation is common in HBsAg-positive patients undergoing autologous HCT. A high HBV DNA level (>10(5) copies/mL) was the most important risk factor for HBV reactivation, and its lowering by administration of nucleoside analogues before transplantation should be considered. PMID- 11895764 TI - High incidence of Kaposi sarcoma-associated herpesvirus-related non-Hodgkin lymphoma in patients with HIV infection and multicentric Castleman disease. AB - Multicentric Castleman disease (MCD) is a distinct type of lymphoproliferative disorder associated with inflammatory symptoms and interleukin 6 (IL-6) dysregulation. In the context of human immunodeficiency virus (HIV) infection, MCD is associated with Kaposi sarcoma-associated herpesvirus, also called human herpesvirus type 8 (KSHV/HHV8). Within a prospective cohort study on 60 HIV infected patients with MCD, and a median follow-up period of 20 months, 14 patients developed KSHV/HHV8-associated non-Hodgkin lymphoma (NHL): 3 "classic" KSHV/HHV8(+) Epstein-Barr virus-positive (EBV(+)) primary effusion lymphoma (PEL), 5 KSHV/HHV8(+) EBV(-) visceral large cell NHL with a PEL-like phenotype, and 6 plasmablastic lymphoma/leukemia (3/3 KSHV/HHV8(+) EBV(-)). The NHL incidence observed in this cohort study (101/1000 patient-years) is about 15-fold what is expected in the general HIV(+) population. MCD-associated KSHV/HHV8(+) NHL fell into 2 groups, suggesting different pathogenesis. The plasmablastic NHL likely represents the expansion of plasmablastic microlymphoma from the MCD lesion and progression toward aggressive NHL. In contrast, the PEL and PEL-like NHL may implicate a different original infected cell whose growth is promoted by the cytokine-rich environment of the MCD lesions. PMID- 11895765 TI - The effects of postponing prophylactic treatment on long-term outcome in patients with severe hemophilia. AB - To prevent hemophilic arthropathy, prophylactic treatment of children with severe hemophilia should be started before joint damage has occurred. However, treatment is expensive, and the burden of regular venipunctures in young children is high. With the aim of providing information on starting prophylaxis on the basis of individual patient characteristics, the effect of postponing prophylaxis on long term arthropathy was studied in a cohort of 76 patients with severe hemophilia born between 1965 and 1985. The median age at first joint bleed was 2.2 years (range, 0.2-5.8). Prophylaxis was started at a median age of 6 years (interquartile range [IQR], 4-9), and the median annual clotting factor use on prophylaxis was 1750 IU/kg/y (31 IU/kg/wk). Hemophilic arthropathy was measured by the Pettersson score (maximum, 78 points). At a median age of 19 years, the median Pettersson score was 7 points (IQR, 0-17). After 2 decades of follow-up, the Pettersson score was 8% higher (95% confidence interval, 1%-16%) for every year prophylaxis was postponed after the first joint bleed. This effect was independent of age at Pettersson score, age at first joint bleed, and prophylactic dose used. In conclusion, most patients have their first joint bleed after the age of 2 years. Patients who start prophylaxis soon after the first joint bleed show little arthropathy in adulthood. The longer the start of prophylaxis is postponed after the first joint bleed, the higher the risk of developing arthropathy. PMID- 11895766 TI - Efficient gene transfer into human primary blood lymphocytes by surface engineered lentiviral vectors that display a T cell-activating polypeptide. AB - In contrast to oncoretroviruses, lentiviruses such as human immunodeficiency virus 1 (HIV-1) are able to integrate their genetic material into the genome of nonproliferating cells that are metabolically active. Likewise, vectors derived from HIV-1 can transduce many types of nonproliferating cells, with the exception of some particular quiescent cell types such as resting T cells. Completion of reverse transcription, nuclear import, and subsequent integration of the lentivirus genome do not occur in these cells unless they are activated via the T cell receptor (TCR) or by cytokines or both. However, to preserve the functional properties of these important gene therapy target cells, only minimal activation with cytokines or TCR-specific antibodies should be performed during gene transfer. Here we report the characterization of HIV-1-derived lentiviral vectors whose virion surface was genetically engineered to display a T cell-activating single-chain antibody polypeptide derived from the anti-CD3 OKT3 monoclonal antibody. Interaction of OKT3 IgGs with the TCR can activate resting peripheral blood lymphocytes (PBLs) by promoting the transition from G(0) to G(1) phases of the cell cycle. Compared to unmodified HIV-1-based vectors, OKT3-displaying lentiviral vectors strongly increased gene delivery in freshly isolated PBLs by up to 100-fold. Up to 48% transduction could be obtained without addition of PBL activation stimuli during infection. Taken together, these results show that surface-engineered lentiviral vectors significantly improve transduction of primary lymphocytes by activating the target cells. Moreover these results provide a proof of concept for an approach that may have utility in various gene transfer applications, including in vivo gene delivery. PMID- 11895767 TI - Role of the docking protein Gab2 in beta(1)-integrin signaling pathway-mediated hematopoietic cell adhesion and migration. AB - Gab2, a newly identified pleckstrin homology domain-containing docking protein, is a major binding protein of SHP-2 tyrosine phosphatase in interleukin (IL)-3 stimulated hematopoietic cells. Its signaling mechanism remains largely unknown. We report here an important regulatory role for Gab2 in beta(1) integrin signaling pathway that mediates hematopoietic cell adhesion and migration. Cross linking of the beta(1) integrin on Ba/F3 cells induced rapid tyrosine phosphorylation of Gab2 and its association with Syk kinase, SHP-2 phosphatase, and the p85 subunit of phosphatidylinositol (PI)-3 kinase. In addition, Gab2 was also constitutively associated with SHP-1 phosphatase via its C-terminal Src homology 2 domain. Overexpression of the pleckstrin homology domain or a mutant Gab2 molecule lacking SHP-2 binding sites resulted in significant reductions in Ba/F3 cell adhesion and migration. Biochemical analyses revealed that enforced expression of Gab2 mutant molecules dramatically reduced beta(1)-integrin ligation-triggered PI3 kinase activation, whereas Erk kinase activation remained unaltered. Furthermore, transduction of primary hematopoietic progenitor cells from viable motheaten mice with these mutant Gab2 molecules also significantly ameliorated their enhanced migration capacity associated with the SHP1 gene mutation. Taken together, these results suggest an important signaling role for Gab2 in regulating hematopoietic cell adhesion and migration. PMID- 11895769 TI - Notch1 activation increases hematopoietic stem cell self-renewal in vivo and favors lymphoid over myeloid lineage outcome. AB - Hematopoietic stem cells sequentially pass through a series of decision points affecting self-renewal or lineage-specific differentiation. Notch1 receptor is a known modulator of lineage-specific events in hematopoiesis that we assessed in the context of in vivo stem cell kinetics. Using RAG-1(-/-) mouse stems cells, we documented increased stem cell numbers due to decreased differentiation and enhanced stem cell self-renewal induced by Notch1. Unexpectedly, preferential lymphoid over myeloid lineage commitment was noted when differentiation occurred. Therefore, Notch1 affects 2 decision points in stem cell regulation, favoring self-renewal over differentiation and lymphoid over myeloid lineage outcome. Notch1 offers an attractive target for stem cell manipulation strategies, particularly in the context of immunodeficiency and acquired immunodeficiency syndrome. PMID- 11895768 TI - Macrophage colony stimulating factor modulates the development of hematopoiesis by stimulating the differentiation of endothelial cells in the AGM region. AB - Definitive hematopoietic stem cells arise in the aorta-gonad-mesonephros (AGM) region from hemangioblasts, common precursors for hematopoietic and endothelial cells. Previously, we showed that multipotential hematopoietic progenitors and endothelial cells were massively produced in primary culture of the AGM region in the presence of oncostatin M. Here we describe a role for macrophage-colony stimulating factor (M-CSF) in the development of hematopoietic and endothelial cells in AGM culture. The number of hematopoietic progenitors including multipotential cells was significantly increased in the AGM culture of op/op embryos. The addition of M-CSF to op/op AGM culture decreased colony-forming unit (CFU)-GEMM, granulocyte macrophage-CFU, and erythroid-CFU, but it increased CFU M. On the other hand, the number of cells expressing endothelial markers, vascular endothelial-cadherin, intercellular adhesion molecule 2, and Flk-1 was reduced in op/op AGM culture. The M-CSF receptor was expressed in PCLP1(+)CD45(-) cells, the precursors of endothelial cells, and M-CSF up-regulated the expression of more mature endothelial cell markers-VCAM-1, PECAM-1, and E-selectin-in PCLP1(+)CD45(-) cells. These results suggest that M-CSF modulates the development of hematopoiesis by stimulating the differentiation of PCLP-1(+)CD45(-) cells to endothelial cells in the AGM region. PMID- 11895770 TI - Retinoid signaling regulates primitive (yolk sac) hematopoiesis. AB - It is known from nutritional studies that vitamin A is an important factor for normal hematopoiesis, though it has been difficult to define its precise role. The vitamin A-deficient (VAD) quail embryo provides an effective ligand "knockout" model for investigating the function of retinoids during development. The VAD embryo develops with a significant reduction in erythroid cells, which has not been noted previously. Activation of the primitive erythroid program and early expression of the erythroid marker GATA-1 occurs, though GATA-1 levels eventually decline, consistent with the erythropoietic and hemoglobin deficits. However, from its early stages, the GATA-2 gene fails to be expressed normally in VAD embryos. The bone morphogenetic protein (BMP)-signaling pathway regulates GATA-2, and BMP4 expression becomes reduced in the caudal embryonic region of VAD embryos. Adding BMP4 to cultured VAD-derived explants rescues the production of erythroid cells, whereas normal embryos cultured in the presence of the BMP antagonist noggin are defective in primitive hematopoiesis. We find that cell clusters of primitive blood islands undergo an inappropriate program of apoptosis in the VAD embryo, which can explain the deficit in differentiated primitive blood cells. We propose that vitamin A-derived retinoids are required for normal yolk sac hematopoiesis and that an embryonic retinoid-BMP-GATA-2 signaling pathway controls progenitor cell survival relevant to primitive hematopoiesis. PMID- 11895771 TI - Early hematopoietic reconstitution after clinical stem cell transplantation: evidence for stochastic stem cell behavior and limited acceleration in telomere loss. AB - Our inability to purify hematopoietic stem cells (HSCs) precludes direct study of many aspects of their behavior in the clinical hematopoietic stem cell transplantation (HSCT) setting. We indirectly assessed stem/progenitor cell behavior in the first year after HSCT by examining changes in neutrophil telomere length, X-inactivation ratios, and cycling of marrow progenitors in 25 fully engrafted allogeneic HSCT recipients. Donors were sampled once and recipients at engraftment and 2 to 6 months and 12 months after HSCT. Telomere length was measured by an in-gel hybridization technique, X-inactivation ratios were measured by the human androgen receptor assay, and cell cycle status was determined by flow cytometric analysis of pyronin Y- and Hoechst 33342-stained CD34(+)CD90(+) and CD34(+)CD90(-) marrow cells. Compared with their donors, recipients' telomeres were shortened at engraftment (-424 base pairs [bp]; P <.0001), 6 months (-495 bp; P =.0001) after HSCT, and 12 months after HSCT (-565 bp; P <.0001). There was no consistent pattern of change in telomere length from 1 to 12 months after HSCT; marked, seemingly random, fluctuations were common. In 11 of 11 informative recipients, donor X-inactivation ratios were faithfully reproduced and maintained. The proportion of CD34(+)CD90(+) progenitors in S/G(2)/M was 4.3% in donors, 15.7% at 2 to 6 months (P <.0001) after HSCT, and 11.5% at 12 months after HSCT (P <.0001, versus donors; P =.04, versus 2-6 months). Cycling of CD34(+) CD90(-) progenitors was largely unchanged. We infer that (1) HSCT-induced accelerated telomere loss is temporary and unlikely to promote graft failure or clonal hematopoietic disorders and (2) the striking fluctuations in telomere length and variation in pattern of telomere loss reflect stochastic determination of HSC fate after HSCT. PMID- 11895772 TI - Vascular endothelial growth factor receptor Flt-1 negatively regulates developmental blood vessel formation by modulating endothelial cell division. AB - Mice lacking the vascular endothelial growth factor (VEGF) receptor flt-1 die of vascular overgrowth, and we are interested in how flt-1 normally prevents this outcome. Our results support a model whereby aberrant endothelial cell division is the cellular mechanism resulting in vascular overgrowth, and they suggest that VEGF-dependent endothelial cell division is normally finely modulated by flt-1 to produce blood vessels. Flt-1(-/-) embryonic stem cell cultures had a 2-fold increase in endothelial cells by day 8, and the endothelial cell mitotic index was significantly elevated before day 8. Flt-1 mutant embryos also had an increased endothelial cell mitotic index, indicating that aberrant endothelial cell division occurs in vivo in the absence of flt-1. The flt-1 mutant vasculature of the cultures was partially rescued by mitomycin C treatment, consistent with a cell division defect in the mutant background. Analysis of cultures at earlier time points showed no significant differences until day 5, when flt-1 mutant cultures had increased beta-galactosidase(+) cells, indicating that the expansion of flt-1 responsive cells occurs after day 4. Mitomycin C treatment blocked this early expansion, suggesting that aberrant division of angioblasts and/or endothelial cells is a hallmark of the flt-1 mutant phenotype throughout vascular development. Consistent with this model is the finding that expansion of platelet and endothelial cell adhesion molecule(+) and VE cadherin(+) vascular cells in the flt-1 mutant background first occurs between day 5 and day 6. Taken together, these data show that flt-1 normally modulates vascular growth by controlling the rate of endothelial cell division both in vitro and in vivo. PMID- 11895773 TI - The NFY transcription factor functions as a repressor and activator of the von Willebrand factor promoter. AB - Human von Willebrand factor (VWF) gene sequences -487 to +247 function as an endothelial-specific promoter in vitro. Analysis of the activation mechanism of the VWF promoter has resulted in the identification of a number of cis-acting elements and trans-acting factors that regulate its activity. The GATA and Ets transcription factors were shown to function as activators of transcription, whereas NF1 and Oct1 were shown to repress transcription. We have reported the presence of another repressor element in exon 1 that interacted with a protein complex designated "R." In the absence of NF1 binding, inhibition of this interaction resulted in promoter activation in nonendothelial cells. We have now identified the "R" protein complex as the NFY transcription factor. Using DNA methylation interference assay and base substitution mutation analysis, we show that NFY interacts with a novel DNA sequence corresponding to nucleotides +226 to +234 in the VWF promoter that does not conform to the consensus NFY binding sequence CCAAT. The VWF gene does contain a CCAAT element that is located downstream of the TATA box and we show that the NFY factor also interacts with this CCAAT element. Using antibodies specific against the A, B, and C subunits of NFY, we demonstrate that the NFY complexes interacting with the CCAAT sequence have a composition similar to that of the repressor binding to the first exon sequences. The results of mutation analysis and transfection studies demonstrated that the interaction of NFY with the upstream CCAAT element is required for VWF promoter activation. Based on these results, we hypothesize that NFY can function both as a repressor and activator of transcription and its function may be modulated through its DNA binding sequences. PMID- 11895774 TI - Adducin in platelets: activation-induced phosphorylation by PKC and proteolysis by calpain. AB - Adducins are a family of cytoskeletal proteins encoded by 3 genes (alpha, beta, and gamma). Platelets express alpha and gamma adducins, in contrast to red blood cells that express alpha and beta adducins. During platelet activation with thrombin, calcium ionophore A23187, or phorbol 12-myristate 13-acetate, alpha and gamma adducins were phosphorylated by protein kinase C (PKC) as detected by an antibody specific for a phosphopeptide sequence in the highly conserved carboxy terminus. Platelet activation also led to adducin proteolysis; inhibition by calpeptin suggests that the protease was calpain. The kinase inhibitor staurosporine inhibited PKC phosphorylation of adducin and also inhibited proteolysis of adducin. Experiments with recombinant alpha adducin demonstrated that the PKC-phosphorylated form was proteolyzed at a significantly faster rate than the unphosphorylated form. The concentration of adducin in platelets was estimated at 6 microM, similar to the concentration of capping protein. Fractionation of platelets into high-speed supernatant (cytosol) and pellet (membrane and cytoskeleton) revealed a shift of PKC-phosphorylated adducin to the cytosol during platelet activation. Platelet aggregation detected turbidometrically was decreased in the presence of staurosporine and was completely inhibited by calpeptin. Thrombin-induced changes in morphology were assessed by confocal microscopy with fluorescein phalloidin and were not prevented by staurosporine or calpeptin. Our results suggest that regulation of adducin function by PKC and calpain may play a role in platelet aggregation. PMID- 11895775 TI - Human single chain antibodies against heparin: selection, characterization, and effect on coagulation. AB - Heparin, located in mast cells and basophilic granulocytes, is widely used as an anticoagulant. It belongs to a class of linear polysaccharides called glycosaminoglycans (GAGs). Using phage display technology, we have selected 19 unique human antiheparin antibodies. Some antibodies react almost exclusively with heparin, others also react with the structurally related heparan sulfate, and some with chondroitin sulfate. In all cases, sulfate groups are essential for binding. For activity of some antibodies, O-sulfation is more important than N sulfation. Antibodies are reactive with heparin in mast cells. Each antibody showed a defined staining pattern on cryosections of rat kidney, pancreas, and testis. Enzymatic digestion with glycosidases on tissue sections further indicated that the antibodies are specific for GAGs. All antibodies recognize a unique epitope. The effect of the antibodies on heparin as an anticoagulant was also studied. There were 3 antibodies that were very effective inhibitors of heparin action in the activated partial thromboplastin time (APTT) clotting assay, and their effect was related to the amount of heparin bound. Some antibodies reacted strongly with the pentasaccharide, which interacts with antithrombin III. The human antibodies selected represent unique tools to study the structure, location, and function of heparin and related GAGs, and some may be used as blocking agents. PMID- 11895776 TI - A hereditary bleeding disorder of dogs caused by a lack of platelet procoagulant activity. AB - We have discovered a novel canine hereditary bleeding disorder with the characteristic features of Scott syndrome, a rare defect of platelet procoagulant activity. Affected dogs were from a single, inbred colony and experienced clinical signs of epistaxis, hyphema, intramuscular hematoma, and prolonged bleeding with cutaneous bruising after surgery. The hemostatic abnormalities identified were restricted to tests of platelet procoagulant activity, whereas platelet count, platelet morphology under light microscopy, bleeding time, clot retraction, and platelet aggregation and secretion in response to thrombin, collagen, and adenosine diphosphate stimulation were all within normal limits. Washed platelets from the affected dogs demonstrated approximately twice normal clotting times in a platelet factor 3 availability assay and, in a prothrombinase assay, generated only background levels of thrombin in response to calcium ionophore, thrombin, or combined thrombin plus collagen stimulation. While platelet phospholipid content was normal, flow cytometric analyses revealed diminished phosphatidylserine exposure and a failure of microvesiculation in response to calcium ionophore, thrombin, and collagen stimulation. Pedigree studies indicate a likely homozygous recessive inheritance pattern of the defect. These findings confirm the importance of platelet procoagulant activity for in vivo hemostasis and provide a large animal model for studying agonist-induced signal transduction, calcium mobilization, and effector pathways involved in the late platelet response of transmembrane phospholipid movement and membrane vesiculation. PMID- 11895777 TI - Role of the Src family kinase Lyn in TxA2 production, adenosine diphosphate secretion, Akt phosphorylation, and irreversible aggregation in platelets stimulated with gamma-thrombin. AB - Members of the Src family of kinases are abundant in platelets. Although their localization is known, their role(s) in platelet function are not well understood. Lyn is a Src-family kinase that participates in signal transduction pathways elicited by collagen-related peptide; it has also been implicated through biochemical studies in the regulation of von Willebrand factor signaling. Here, we provide evidence that Lyn plays a role in gamma-thrombin activation of platelets. Unlike the wild-type platelets, platelets from Lyn-deficient mice do not undergo irreversible aggregation, produce thromboxane A2, or secrete adenosine diphosphate in response to submaximal gamma-thrombin concentrations that cause secretion-dependent irreversible aggregation. Phosphorylation of Akt, a downstream effector of phosphatidylinositol 3-kinase, also requires a higher concentration of gamma-thrombin in Lyn-deficient platelets than in wild-type platelets. These findings demonstrate that Lyn signaling is required for thrombin induction of secretion-dependent platelet aggregation. Specifically, Lyn is required under these conditions to enable thrombin-induced TxA2 production and adenosine diphosphate secretion, necessary steps in secretion-dependent platelet aggregation. PMID- 11895778 TI - Factor XI deficiency in French Basques is caused predominantly by an ancestral Cys38Arg mutation in the factor XI gene. AB - Inherited factor XI deficiency is an injury-related bleeding disorder that is rare in most populations except for Jews, in whom 2 mutations, a stop mutation in exon 5 (type II) and a missense mutation in exon 9 (type III), predominate. Recently, a cluster of 39 factor XI-deficient patients was described in the Basque population of Southwestern France. In this study, we determined the molecular basis of factor XI deficiency in 16 patients belonging to 12 unrelated families of French Basque origin. In 8 families, a nucleotide 209T>C transition in exon 3 was detected that predicts a Cys38Arg substitution. Four additional novel mutations in the factor XI gene, Cys237Tyr, Tyr493His, codon 285delG, and IVS6 + 3A>G, were identified in 4 families. Expression studies showed that Cys38Arg and Cys237Tyr factor XI were produced in transfected baby hamster kidney cells, but their secretion was impaired. Cells transfected with Tyr493His contained reduced amounts of factor XI and displayed decreased secretion. A survey of 206 French Basque controls for Cys38Arg revealed that the prevalence of the mutant allele was 0.005. Haplotype analysis based on the study of 10 intragenic polymorphisms was consistent with a common ancestry (a founder effect) for the Cys38Arg mutation. PMID- 11895779 TI - Impact of antithrombin deficiency in thrombogenesis: lipopolysaccharide and stress-induced thrombus formation in heterozygous antithrombin-deficient mice. AB - Antithrombin (AT) deficiency is an autosomal disorder associated with venous thromboembolism. However, a diagnosis of homozygous AT deficiency is seldom made. Most patients are heterozygous and have approximately 50% AT activities, and they are at higher risk for the development of thromboembolism. Through gene targeting we generated AT-deficient mice and previously reported that completely AT deficient mice could not survive the prenatal period because of extensive thrombosis in the myocardium and liver sinusoids. In contrast, heterozygous AT deficient mice with 50% AT activities have not shown spontaneous thromboembolic episodes. To demonstrate a thrombotic tendency in heterozygous AT deficiency, we challenged heterozygous AT-deficient mice (AT+/- mice) with the administration of lipopolysaccharide (LPS) or with restraint stress by immobilization. LPS injection markedly induced fibrin deposition in the kidney glomeruli, myocardium, and liver sinusoids in AT+/- mice compared with wild-type mice (AT+/+ mice). Restraint stress tests were performed by placing mice in 50-mL conical centrifuge tubes for 20 hours. Fibrin deposition was observed in the kidney of AT+/+ and AT+/- mice, but AT+/- mice exhibited more extensive fibrin deposition than AT+/+ mice. After prophylactic administration of human AT concentrates to increase plasma AT activities of AT+/- mice, LPS-induced fibrin deposition was effectively prevented. These results suggest that heterozygous AT deficiency is significantly associated with a tendency toward thrombosis formation in the kidney. The AT+/- mouse thus is a useful model for studying the effect of environmental or genetic risk factors on thrombogenesis. PMID- 11895780 TI - Differential surrogate light chain expression governs B-cell differentiation. AB - Surrogate light chain expression during B lineage differentiation was examined by using indicator fluorochrome-filled liposomes in an enhanced immunofluorescence assay. Pro-B cells bearing surrogate light chain components were found in mice, but not in humans. A limited subpopulation of relatively large pre-B cells in both species expressed pre-B cell receptors. These cells had reduced expression of the recombinase activating genes, RAG-1 and RAG-2. Their receptor-negative pre B cell progeny were relatively small, expressed RAG-1 and RAG-2, and exhibited selective down-regulation of VpreB and lambda5 expression. Comparative analysis of the 2 pre-B cell subpopulations indicated that loss of the pre-B cell receptors from surrogate light chain gene silencing was linked with exit from the cell cycle and light chain gene rearrangement to achieve B-cell differentiation. PMID- 11895781 TI - CD4(+) and CD8(+) anergic T cells induced by interleukin-10-treated human dendritic cells display antigen-specific suppressor activity. AB - Interleukin-10 (IL-10)-treated dendritic cells (DCs) induce an alloantigen- or peptide-specific anergy in various CD4(+) and CD8(+) T-cell populations. In the present study, we analyzed whether these anergic T cells are able to regulate antigen-specific immunity. Coculture experiments revealed that alloantigen specific anergic CD4(+) and CD8(+) T cells suppressed proliferation of syngeneic T cells in a dose-dependent manner. The same effect was observed when the hemagglutinin-specific CD4(+) T-cell clone HA1.7 or tyrosinase-specific CD8(+) T cells were cocultured with anergic T cells of the same specificity. Anergic T cells did not induce an antigen-independent bystander inhibition. Suppression was dependent on cell-to-cell contact between anergic and responder T cells, required activation by antigen-loaded DCs, and was not mediated by supernatants of anergic T cells. Furthermore, anergic T cells displayed an increased extracellular and intracellular expression of cytotoxic T-lymphocyte antigen (CTLA)-4 molecules, and blocking of the CTLA-4 pathway restored the T-cell proliferation up to 70%, indicating an important role of the CTLA-4 molecule in the suppressor activity of anergic T cells. Taken together, our experiments demonstrate that anergic T cells induced by IL-10-treated DCs are able to suppress activation and function of T cells in an antigen-specific manner. Induction of anergic T cells might be exploited therapeutically for suppression of cellular immune responses in allergic or autoimmune diseases with identified (auto) antigens. PMID- 11895782 TI - Expression of ABO or related antigenic carbohydrates on viral envelopes leads to neutralization in the presence of serum containing specific natural antibodies and complement. AB - No definitive biologic function has been associated with the human ABO histo blood group polymorphism, or any other terminal carbohydrate differences within or between closely related species. We have experimentally addressed the question of whether viral particles can become glycosylated as determined by the glycosylation (eg, ABO) status of the producer cell and as a result be affected by human serum containing specific natural antibodies (NAbs). Measles virus was produced in cells transfected with cDNA encoding, either human A-transferase, B transferase, an inactive "O-transferase," or a pig alpha1-3galactosyltransferase (alpha1-3GT) synthesizing the Galalpha1-3Gal structure. The viruses were shown to carry the same ABO structures as the cells; that is, A but not B if produced in A type cells, and B but not A if produced in B-type cells. Only O was detected on the virus produced from O-type cells, whereas reduced amounts of O appeared on the A- and B-type viral particles. In addition, the Galalpha1-3Gal structure was transferred onto measles only when grown in human cells expressing this structure. When subjected to human preimmune sera, the A-type, the B-type, and the Galalpha1-3Gal viral particles were partially neutralized in a complement dependent manner. However, the O-type or the Galalpha1-3Gal-negative viral particles were not neutralized. The neutralization appeared to be mediated by specific NAb, as judged by specific inhibition using synthetic A and Galalpha1 3Gal oligosaccharides. Such viral glycosylation may thus partly explain why the ABO antigens and other similar intraspecies as well as interspecies polymorphic carbohydrates have evolved and been maintained over long evolutionary periods. PMID- 11895783 TI - Ex vivo development of functional human lymph node and bronchus-associated lymphoid tissue. AB - We introduce a novel in vivo model of human mucosal immunity, based on the implantation of human fetal bronchial mucosa and autologous peribronchial lymph node (PLN) in the severe combined immunodeficiency (SCID) mouse. In the SCID host, human fetal bronchi implanted alone retain macrophages and mast cells but lose T cells. In contrast, fetal bronchi co-implanted with PLN contain, in addition to macrophages and mast cells, numerous T cells and B cells, often clustered in intramucosal bronchus-associated lymphoid tissue (BALT). Functionally, bronchus-PLN cografts are able to mount robust alphabeta and gammadelta T-cell-mediated immune responses to Pseudomonas aeruginosa and 3,4 epoxy-3-methyl-1-butyl-diphosphate challenges. No other autologous lymphoid organ (bone marrow, thymus, liver) allows for BALT development in co-implanted bronchi, which suggests special ontogenetic and functional relations between extramucosal PLN and intramucosal BALT. Overall, the bronchus-PLN cograft appears as a promising model for human bronchial immune development and function. Our study is the first to document long-term ex vivo maintenance of functional human lymph nodes as native appendices to mucosal tissue. Our results, therefore, suggest a simple strategy for developing similar experimental models of human immune function in other mucosae. PMID- 11895785 TI - Restricted T-cell receptor beta-chain usage by T cells autoreactive to beta(2) glycoprotein I in patients with antiphospholipid syndrome. AB - We recently identified CD4(+) T cells that are autoreactive to beta(2) glycoprotein I (beta(2)GPI) and that promote antiphospholipid antibody production in patients with antiphospholipid syndrome (APS). In this study, T-cell receptor (TCR) beta chains of beta(2)GPI-reactive T cells were examined in 8 beta(2)GPI responders, including 5 patients with APS and 3 healthy subjects, using polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) analysis combined with in vitro stimulation of peripheral blood T cells with recombinant beta(2)GPI. The TCR Vbeta segments that expanded oligoclonally after stimulation with beta(2)GPI varied among responders, but the Vbeta7 and Vbeta8 segments were commonly detected in 6 and 4 beta(2)GPI-responders, respectively. Analysis of the complementarity-determining region 3 sequence of beta(2)GPI reactive T cells revealed limited diversity, and all Vbeta7(+) TCRs had an amino acid motif of TGxxN/Q or minor variations. The Vbeta8(+) TCRs had another motif, PxAxxD/E. Surprisingly, an identical Vbeta7(+) TCRbeta chain was used by beta(2)GPI-reactive T cells in 3 patients with APS. There was no apparent difference in the TCRbeta usage between APS patients and healthy responders. Some of the Vbeta7(+) TCRs with the TGxxN/Q motif detected by PCR-SSCP analysis were also used by beta(2)GPI-specific CD4(+) T-cell clones responsive to an immunodominant epitope containing the major phospholipid-binding site. Depletion of Vbeta7(+) or Vbeta8(+) T cells from the peripheral blood mononuclear cell cultures significantly inhibited in vitro anti-beta(2)GPI antibody production in response to beta(2)GPI. Our results indicate preferential usage of TCRbeta chains by beta(2)GPI-reactive T cells. These TCRbeta chains can be reasonable targets for TCR-based immunotherapy for patients with APS. PMID- 11895784 TI - Human CD38 and CD16 are functionally dependent and physically associated in natural killer cells. AB - CD38, a surface glycoprotein of unrestricted lineage, is an ectoenzyme (adenosine diphosphate [ADP] ribosyl cyclase/cyclic ADP-ribose hydrolase) that regulates cytoplasmic calcium. The molecule also performs as a receptor, modulating cell cell interactions and delivering transmembrane signals, despite showing a structural ineptitude to the scope. CD38 ligation by agonistic monoclonal antibodies induced signals leading to activation of the lytic machinery of natural killer (NK) cells from adults; similar signals could not be reproduced in YT and NKL, 2 CD16(-) human NK-like lines. It was hypothesized that CD38 establishes a functional cooperation with professional signaling molecules of the NK cell surface. The present work answers the question about the molecule exploited by CD38 for signaling in NK cells, using as a model CD16(-) NK lines genetically corrected for CD16 expression. Our results indicate that a functional CD16 molecule is a necessary and sufficient requisite for CD38 to control an activation pathway, which includes calcium fluxes, tyrosine phosphorylation of ZAP70 and mitogen-activated protein kinase, secretion of interferon-gamma, and cytotoxic responses. Fluorescence resonance energy transfer and cocapping experiments also showed a surface proximity between CD38 and CD16. These results were confirmed by using the NKL cell line, in which CD16(+) and CD16(-) variants were obtained without genetic manipulation. Together, our findings show CD38 to be a unique receptor molecule that cannot signal by itself but whose receptor function is rescued by functional and physical associations with a professional signaling structure that varies according to lineage and environment. This molecule is CD16 in NK cells. PMID- 11895786 TI - Persistent numbers of tetramer+ CD8(+) T cells, but loss of interferon-gamma+ HIV specific T cells during progression to AIDS. AB - Although CD8(+) T cells initially suppress human immunodeficiency virus (HIV) replication, cytotoxic T-cell precursor frequencies eventually decline and fail to prevent disease progression. In a longitudinal study including 16 individuals infected with HIV-1, we studied both the number and function of HIV-specific CD8(+) T cells by comparing HLA-peptide tetramer staining and peptide-induced interferon-gamma (IFN-gamma) production. Numbers of IFN-gamma-producing T cells declined during progression to acquired immunodeficiency syndrome (AIDS), whereas the number of tetramer+ T cells in many individuals persisted at high frequencies. Loss of IFN-gamma-producing T cells correlated with declining CD4(+) T-cell counts, consistent with the need of CD4(+) T-cell help in maintaining adequate CD8(+) T-cell function. These data indicate that the loss of HIV specific CD8(+) T-cell activity is not due to physical depletion, but is mainly due to progressively impaired function of HIV-specific CD8(+) T cells. PMID- 11895787 TI - Immunization against murine multiple myeloma with fusions of dendritic and plasmacytoma cells is potentiated by interleukin 12. AB - Fusions of cancer cells and dendritic cells (DCs) are effective in the treatment of animal tumor models and patients with metastatic renal carcinoma. In this study, we have fused DCs with mouse 4TOO plasmacytoma cells. The results demonstrate that vaccination of mice with the fusion cells (FC/4TOO) is associated with induction of antitumor humoral and cytotoxic T lymphocyte (CTL) responses. Immunization with FC/4TOO cells protected mice against tumor challenge. In addition, treatment of established multiple myeloma with FC/4TOO cells was associated with prolongation of survival but not with eradication of disease. As interleukin (IL)-12 potentiates the induction of immune responses, recombinant mouse IL-12 was administered with the FC/4TOO vaccine. Treatment of mice with FC/4TOO and IL-12 was associated with increased CTL activity and T-cell proliferation responses. Treatment with FC/4TOO and IL-12 also resulted in eradication of established disease. These findings demonstrate that immunization with FC/4TOO fusion cells and IL-12 potentiates antitumor immunity and the treatment of murine multiple myeloma. PMID- 11895788 TI - CD43 polarization in unprimed T cells can be dissociated from raft coalescence by inhibition of HMG CoA reductase. AB - Movement of T-lymphocyte cell surface CD43 is associated with both antigen activation of T-cell clones and chemokine induction of T-lymphocyte motility. Here, we demonstrate that CD43 movement away from the site of T-cell receptor ligation occurs in unprimed CD4(+) T cells as well as T-cell clones. The T-cell receptor (TCR)-dependent movement of CD43 in unprimed T cells is associated with a polarized morphology and CD43 accumulation at the uropods of the cells, unlike that reported for primed T cells. The polarization of CD43 has a requirement for Src kinases and occurs in conjunction with lipid raft coalescence. Thymocytes and T-cell hybridomas, cells that have altered responses to TCR activation and lack lipid raft coalescence, do not polarize CD43 as readily as unprimed T cells. The movement of CD43 depends on the cholesterol biosynthetic pathway enzyme 3-hydroxy 3-methylglutaryl coenzyme A (HMG CoA) reductase. Blockade of this enzyme can specifically prevent CD43 redistribution without affecting cell shape polarization. The likely mechanism of this alteration in CD43 redistribution is through decreased protein prenylation because the cholesterol-dependent lipid rafts still coalesce on activation. These findings suggest that the polarization of cell shape, lipid raft coalescence, and CD43 redistribution on T-cell activation have signaling pathway distinctions. Dissecting out the relationships between various stages of molecular redistribution and lymphocyte activation may facilitate fine-tuning of immunologic responses. PMID- 11895789 TI - A cryptic t(5;11)(q35;p15.5) in 2 children with acute myeloid leukemia with apparently normal karyotypes, identified by a multiplex fluorescence in situ hybridization telomere assay. AB - The identification of specific chromosome abnormalities in acute myeloid leukemia (AML) is important for the stratification of patients into the appropriate treatment protocols. However, a significant proportion of diagnostic bone marrow karyotypes in AML is reported as normal by conventional cytogenetic analysis and it is suspected that these karyotypes may conceal the presence of diagnostically significant chromosome rearrangements. To address this question, we have developed a novel 12-color fluorescence in situ hybridization (FISH) assay for telomeric rearrangements (termed M-TEL), which uses an optimized set of chromosome-specific subtelomeric probes. We report here the application of the M TEL assay to 69 AML cases with apparently normal karyotypes or an isolated trisomy. Of the 69 cases examined, 3 abnormalities were identified, all in the normal karyotype group. The first was a t(11;19)(q23;p13), identified in an infant with AML-M4. In 2 other young patients with AML (< 19 years), an apparently identical t(5;11)(q35;p15.5) was identified. Breakpoint mapping by FISH and reverse transcriptase polymerase chain reaction (RT-PCR) analysis confirmed that this was the same t(5;11) as previously identified in 3 children with AML, associated with del(5q) and resulting in the NUP98-NSD1 gene fusion. The t(5;11) was not detected by 24-color karyotyping using multiplex FISH (M FISH), emphasizing the value of screening with subtelomeric probes for subtle translocations. This is the first report of the t(5;11)(q35;p15.5) in association with an apparently normal karyotype, and highlights this as a new, potentially clinically significant chromosome rearrangement in childhood AML. PMID- 11895790 TI - VEGF(165) promotes survival of leukemic cells by Hsp90-mediated induction of Bcl 2 expression and apoptosis inhibition. AB - Similar to endothelial cells (ECs), vascular endothelial growth factor (VEGF) induces Bcl-2 expression on VEGF receptor-positive (VEGFR(+)) primary leukemias and cell lines, promoting survival. We investigated the molecular pathways activated by VEGF on such leukemias, by performing a gene expression analysis of VEGF-treated and untreated HL-60 leukemic cells. One gene to increase after VEGF stimulation was heat shock protein 90 (Hsp90). This was subsequently confirmed at the protein level, on primary leukemias and leukemic cell lines. VEGF increased the expression of Hsp90 by interacting with KDR and activating the mitogen activated protein kinase cascade. In turn, Hsp90 modulated Bcl-2 expression, as shown by a complete blockage of VEGF-induced Bcl-2 expression and binding to Hsp90 by the Hsp90-specific inhibitor geldanamycin (GA). GA also blocked the VEGF induced Hsp90 binding to APAF-1 on leukemic cells, a mechanism shown to inhibit apoptosis. Notably, VEGF blocked the proapoptotic effects of GA, correlating with its effects at the molecular level. Earlier, we showed that in some leukemias, a VEGF/KDR autocrine loop is essential for cell survival, whereas here we identified the molecular correlates for such an effect. We also demonstrate that the generation of a VEGF/VEGFR autocrine loop on VEGFR(+) cells such as ECs, also protected them from apoptosis. Infection of ECs with adenovirus-expressing VEGF resulted in elevated Hsp90 levels, increased Bcl-2 expression, and resistance to serum-free or GA-induced apoptosis. In summary, we demonstrate that Hsp90 mediates antiapoptotic and survival-promoting effects of VEGF, which may contribute to the survival advantage of VEGFR(+) cells such as subsets of leukemias. PMID- 11895791 TI - Molecular follow-up in gastric mucosa-associated lymphoid tissue lymphomas: early analysis of the LY03 cooperative trial. AB - Gastric marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT)-type can regress after anti-Helicobacter pylori treatment. The International Extranodal Lymphoma Study Group, the United Kingdom Lymphoma Group, and the Groupe d'Etude des Lymphomes de l'Adulte have conducted a trial to ascertain whether the addition of chlorambucil is of benefit after anti-H pylori therapy. At the last interim analysis, 105 (55%) of 189 patients had achieved a complete histologic remission after anti-Helicobacter therapy. To further assess the ability of treatment to eradicate the lymphoma clone, we analyzed the gastric biopsies from a subset of the patients by polymerase chain reaction (PCR) targeted to the immunoglobulin heavy chain genes as a molecular marker for minimal residual disease. Sixty-two cases were examined at diagnosis. Fifty-four cases were monoclonal by PCR. Forty-two of these patients achieved histologic complete remission (hCR) after anti-Helicobacter treatment: 34 cases underwent molecular follow-up analysis. Fifteen patients (44%) were in molecular remission with a median follow-up of 2 years after antibiotic treatment and of 1 year after the achievement of hCR. Less than half of the patients with MALT lymphoma can achieve sustained molecular remission after anti-Helicobacter therapy. The presence of molecular disease in the absence of histologic disease does not appear to be associated with histologic relapse, but, given the indolent nature of MALT lymphomas, a longer follow-up is needed. PMID- 11895792 TI - 5-Hydroxytryptamine drives apoptosis in biopsylike Burkitt lymphoma cells: reversal by selective serotonin reuptake inhibitors. AB - Serotonin (5-HT), a well-known neurotransmitter of the central nervous system, has been implicated in diverse aspects of immune regulation. Here we show that 5 HT can efficiently drive programmed cell death in established Burkitt lymphoma (BL) lines that remain faithful to the original biopsy phenotype (group 1). Group 1 BL cells cultured in the presence of 5-HT exhibited marked suppression of DNA synthesis that was accompanied by extensive apoptosis-serotonin-driven apoptosis was complete within 24 hours, was preceded by early caspase activation, and was accompanied by a decline in mitochondrial membrane potential. BL cells that had drifted to a lymphoblastic group 3 phenotype were relatively resistant to these actions of serotonin, and the forced ectopic expression of either bcl-2 or bcl x(L) provided substantial protection from 5-HT-induced apoptosis. 5-HT receptor antagonists (SDZ205-557, granisetron, methysergide) failed to inhibit serotonin induced apoptosis, whereas the selective serotonin reuptake inhibitors (SSRI) fluoxetine (Prozac), paroxetine (Paxil), and citalopram (Celexa)-substantially blocked the monoamine actions. Western blot analysis showed that BL cells expressed protein for the 5-HT transporter, and transport assays confirmed active uptake of serotonin by the cells. Unlike what was suggested for neuronal cells, there was no evidence that intracellular oxidative metabolites were responsible for the 5-HT-induced programmed death of BL cells. These data indicate that serotonin drives apoptosis in biopsylike BL cells after its entry through an active transport mechanism, and they suggest a novel therapeutic modality for Burkitt lymphoma. PMID- 11895793 TI - Spectral karyotyping identifies new rearrangements, translocations, and clinical associations in diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphoma (DLBCL), a histologically well-defined subset of non-Hodgkin lymphoma, is clinically and genetically heterogenous. By G-banding, most cases showed complex hyperdiploid karyotypes and diverse cytogenetic abnormalities that included recurring and nonrecurring translocations, deletions, duplications, and marker chromosomes. While G-banding provided valuable leads to identification of specific rearrangements that enabled gene discovery and clinical correlations, many aberrations remained uncharacterized because of their complexity. The molecular cytogenetic technique spectral karyotyping (SKY), on the other hand, enables complete characterization of all aberrations in a tumor cell karyotype and, hence, precise quantitation of chromosome instability. We report here, for the first time, SKY analysis of a panel of 46 DLBCL cases previously analyzed by G-banding, ascertained at the Memorial Sloan-Kettering Cancer Center. This analysis provided a cytogenetic profile of DLBCL that was characterized by a higher level of instability, qualitatively as well as quantitatively, compared with G-banding. Thus, 551 breakpoints were detected by SKY, in contrast to the 295 by G-banding. Several new recurring breakpoints, translocations, and regions of gain and loss were identified, which included 13 breakpoints not previously identified by G-banding, 10 breakpoints that were underrepresented by G-banding, and 4 previously unrecognized translocations: der(14)t(3;14)(q21;q32), t(1;13)(p32;q14), t(1;7)(q21;q22), and der(6)t(6;8)(q11;q11). We identified new clinical associations involving recurring breakpoints detected by SKY. These studies emphasize the value of SKY analysis for redefinition of chromosomal instability in DLBCL to enhance gene discovery as well as clinical correlation analysis. PMID- 11895794 TI - Acquisition of potential N-glycosylation sites in the immunoglobulin variable region by somatic mutation is a distinctive feature of follicular lymphoma. AB - Most patients with follicular lymphoma (FL) have somatically mutated V genes with intraclonal variation, consistent with location in the germinal center site. Using our own and published sequences, we have investigated the frequency of potential N-glycosylation sites introduced into functional V(H) genes as a consequence of somatic mutation. FL cells were compared with normal memory B cells or plasma cells matched for similar levels of mutation. Strikingly, novel sites were detected in 55 of 70 (79%) patients with FL, compared to 7 of 75 (9%) in the normal B-cell population (P <.001). Diffuse large B-cell lymphoma (DLCL) showed an intermediate frequency (13 of 32 [41%] patients). Myeloma and the mutated subset of chronic lymphocytic leukemia showed frequencies similar to those of normal cells in 5 of 64 (8%) patients and 5 of 40 (13%) patients, respectively. In 3 of 3 random patients with FL, immunoglobulin was expressed as recombinant single-chain Fv in Pichia pastoris, and glycosylation was demonstrated. These findings indicate that N-glycosylation of the variable region may be common in FL and in a subset of DLCL. Most novel sites are located in the complementarity-determining regions. V(H) sequences of nonfunctional V(H) genes contained few sites, arguing for positive selection in FL. One possibility is that the added carbohydrate in the variable region contributes to interaction with elements in the germinal center environment. This common feature of FL may be critical for tumor behavior. PMID- 11895795 TI - Ca(++)-dependent vesicle release from erythrocytes involves stomatin-specific lipid rafts, synexin (annexin VII), and sorcin. AB - Cytosolic Ca(++) induces the shedding of microvesicles and nanovesicles from erythrocytes. Atomic force microscopy was used to determine the sizes of these vesicles and to resolve the patchy, fine structure of the microvesicle membrane. The vesicles are highly enriched in glycosyl phosphatidylinositol-linked proteins, free of cytoskeletal components, and depleted of the major transmembrane proteins. Both types of vesicles contain 2 as-yet-unrecognized red cell proteins, synexin and sorcin, which translocate from the cytosol to the membrane upon Ca(++) binding. In nanovesicles, synexin and sorcin are the most abundant proteins after hemoglobin. In contrast, the microvesicles are highly enriched in stomatin. The membranes of both microvesicles and nanovesicles contain lipid rafts. Stomatin is the major protein of the microvesicular lipid rafts, whereas synexin and sorcin represent the major proteins of the nanovesicular rafts in the presence of Ca(++). Interestingly, the raft proteins flotillin-1 and flotillin-2 are not found in the vesicles but remain in the red cell membrane. These data indicate the presence of different types of lipid rafts in the erythrocyte membrane with distinct fates after Ca(++) entry. Synexin, which is known to be vital to the process of membrane fusion, is suggested to be a key component in the process of vesicle release from erythrocytes. PMID- 11895796 TI - Dehydration response of sickle cells to sickling-induced Ca(++) permeabilization. AB - Interaction of hemoglobin S polymers with the red blood cell (RBC) membrane induces a reversible increase in permeability ("P(sickle)") to (at least) Na(+), K(+), Ca(2+), and Mg(2+). Resulting changes in [Ca(2+)] and [H(+)] in susceptible cells activate 2 transporters involved in sickle cell dehydration, the Ca(2+) sensitive K(+) ("Gardos") channel (K(Ca)) and the acid- and volume-sensitive K:Cl cotransport. We investigated the distribution of P(sickle) expression among deoxygenated sickle cell anemia (SS) RBCs using new experimental designs in which the RBC Ca(2+) pumps were partially inhibited by vanadate, and the cells' dehydration rates were detected as progressive changes in the profiles of osmotic fragility curves and correlated with flow cytometric measurements. The results exposed marked variations in (sickling plus Ca(2+))-induced dehydration rates within populations of deoxygenated SS cells, with complex distributions, reflecting a broad heterogeneity of their P(sickle) values. P(sickle)-mediated dehydration was inhibited by clotrimazole, verifying the role of K(Ca), and also by elevated [Ca(2+)](o), above 2 mM. Very high P(sickle) values occurred with some SS discocytes, which had a wide initial density (osmotic resistance) distribution. Together with its previously shown stochastic nature, the irregular distribution of P(sickle) documented here in discocytes is consistent with a mechanism involving low-probability, reversible interactions between sickle polymers and membrane or cytoskeletal components, affecting only a fraction of the RBCs during each deoxygenation event and a small number of activated pathways per RBC. A higher participation of SS reticulocytes in P(sickle)-triggered dehydration suggests that they form these pathways more efficiently than discocytes despite their lower cell hemoglobin concentrations. PMID- 11895797 TI - Campath-1G causes rapid depletion of circulating host dendritic cells (DCs) before allogeneic transplantation but does not delay donor DC reconstitution. AB - Graft-versus-host disease (GVHD), a major complication after allogeneic transplantation, can be abrogated by the Campath (anti-CD52) monoclonal antibody. The induction of acute GVHD requires host antigens to be presented to donor T cells by antigen-presenting cells (APCs). Recent evidence has suggested that only host APCs can interact with donor T cells in the induction of GVHD. Because CD52 has been reported to be expressed on DCs, we reasoned that pretransplant Campath 1G might have a direct effect on circulating DCs in addition to any effects on donor T cells. Using direct immunostaining, we demonstrated expression of CD52 on DCs and that Campath-1G killed purified DCs in vitro. In vivo Campath also depleted DCs. Twenty-four hours after the first dose of Campath-1G, circulating DCs were reduced by a mean of 79% (range, 44%-96%). By day 0 after 5 doses of Campath-1G and chemoradiotherapy conditioning, DCs became undetectable in 7 of 9 cases, whereas in 6 of 7 patients receiving conditioning therapy without Campath 1G, host DCs were still detectable. The reconstitution of circulating DCs after transplantation was not affected by Campath-1G and in both groups DC1 (CD11c(+)) recovered more rapidly than DC2 (CD11c(-)). Analysis of chimerism confirmed that the DCs recovering after transplantation in patients receiving Campath-1G were of donor origin. We conclude that in vivo Campath-1G causes a rapid depletion of host circulating DCs and that this may, in part, explain the low incidence of acute GVHD. The reconstitution of donor DCs was not delayed, which may be important in preserving immune reconstitution. PMID- 11895798 TI - Infusion of autologous Epstein-Barr virus (EBV)-specific cytotoxic T cells for prevention of EBV-related lymphoproliferative disorder in solid organ transplant recipients with evidence of active virus replication. AB - Epstein-Barr virus (EBV)-associated posttransplantation lymphoproliferative disorders (PTLDs) are a well-recognized complication of immunosuppression in solid organ transplant recipients. The reported therapeutic approaches are frequently complicated by rejection, toxicity, and other infectious pathologies, and overall mortality in patients with unresponsive PTLD remains high. Thus, low toxicity treatment options or, preferably, some form of prophylactic/preemptive intervention are warranted to improve PTLD outcome in this setting. We assessed whether transfer of EBV-specific cytotoxic T lymphocytes (CTLs) generated in vitro from the peripheral blood of allograft recipients receiving immunosuppression could increase EBV-specific killing in vivo without augmenting the probability of graft rejection. Autologous EBV-specific CTLs were generated for 23 patients who were identified as being at risk of developing PTLD through the finding of elevated EBV DNA load. Of the 23 patients, 7 received 1 to 5 infusions of EBV-specific CTLs. CTL transfer was well tolerated, and none of the patients showed any evidence of rejection. An increase of the EBV-specific cytotoxicity was observed after infusion, notwithstanding continuation of immunosuppressive therapy. EBV DNA levels had a 1.5- to 3-log decrease in 5 patients, whereas in the other 2 graft recipients CTL transfer had no apparent stable effect on EBV load. Our data suggest that the infusion of autologous EBV specific CTLs obtained from peripheral blood mononuclear cells recovered at the time of viral reactivation is able to augment virus-specific immune response and to reduce viral load in organ transplant recipients. This approach may, therefore, be safely used as prophylaxis of EBV-related lymphoproliferative disorders in these patients, following a strategy of preemptive therapy guided by EBV DNA levels. PMID- 11895799 TI - Development of pancytopenia with neutralizing antibodies to thrombopoietin after multicycle chemotherapy supported by megakaryocyte growth and development factor. AB - Clinical trials of thrombopoietin (TPO), the central regulator of megakaryocytopoiesis, have revealed few side effects associated with its use. We here report a case of pancytopenia associated with the development of neutralizing antibodies to TPO that occurred in a patient who had undergone multicycle chemotherapy with multiple cycles of subcutaneous administration of pegylated recombinant human megakaryocyte growth and development factor. Samples of the patient's bone marrow showed trilineage hypoplasia with absence of myeloid, erythroid, and megakaryocyte progenitor cells but with elevated endogenous levels of erythropoietin, granulocyte colony-stimulating factor, and stem-cell factor. To our knowledge, this is the first report of an aplastic anemia-like syndrome associated with neutralizing antibodies to TPO. PMID- 11895800 TI - Erythropoietin receptor haploinsufficiency and in vivo interplay with granulocyte macrophage colony-stimulating factor and interleukin 3. AB - Erythropoietin (EPO) and its receptor (EPOR) are critical for definitive erythropoiesis, as mice lacking either gene product die during embryogenesis with severe anemia. Here we demonstrate that mice expressing just one functional allele of the EpoR have lower hematocrits and die more frequently than do wild type littermates on anemia induction. Furthermore, EpoR(+/-) erythroid colony forming unit (CFU-E) progenitors are reduced both in frequency and in responsiveness to EPO stimulation. To evaluate the interaction between EPO and granulocyte-macrophage colony-stimulating factor (GM-CSF) or interleukin 3 (IL 3), GM-CSF(-/-) or IL-3(-/-) mice were interbred with EpoR(+/)(-) mice. Deletion of either GM-CSF or IL-3 also leads to reduction in CFU-E numbers and hematocrits but does not significantly alter steady-state erythroid burst-forming unit numbers. These results suggest EpoR haploinsufficiency and promotion of in vivo erythropoiesis by GM-CSF and IL-3. PMID- 11895801 TI - High frequency of protein Z deficiency in patients with unexplained early fetal loss. AB - The protein Z-protein Z-dependent inhibitor complex is a factor Xa inhibitor. Protein Z deficiencies have recently been described in patients with ischemic stroke. As placenta infarction leads to poor pregnancy outcome, we studied protein Z plasma concentrations in nonthrombotic, nonthrombophilic consecutive patients with unexplained pregnancy wastage. A significant amount of protein Z deficiencies was only found in the early fetal loss group (< 1 mg/L; 44 of 200, P < 10(-4)) and mainly in the case of fetal demise between the beginning of the 10th and the end of the 15th week of gestation (odds ratio, 6.7 [3.1-14.8], P < 10(-3)). These deficiencies were not due to partial vitamin K1 deficiency, and at least some of them were constitutional ones. In women, protein Z deficiency may induce an enhanced risk of severe placental insufficiency soon after the connection of maternal and fetal circulations. PMID- 11895802 TI - O-glycans on human high endothelial CD34 putatively participating in L-selectin recognition. AB - Leukocyte traffic into lymph nodes and sites of inflammation is guided by L selectin. Experiments performed in vitro and with gene-deleted mice suggest that CD34 recognizes L-selectin if decorated by 6-sulfo sialyl Lewis x (sLex) saccharides and the MECA-79 epitope. However, very little is known about glycosylation of human L-selectin ligands. We report here on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiles of N- and O-linked oligosaccharide fractions from human tonsillar endothelial CD34. All detected O-glycans were sialylated; some were also monosulfated or monosulfated and monofucosylated. If a given CD34-glycan may carry all requirements for L-selectin recognition, that is, both 6-sulfo-sLex and MECA-79 epitopes, only one O-glycan fraction, O-9, SA(2)Hex(3)HexNAc(3)- Fuc(1)(SO(3))(1), meets the criteria. A candidate structure is SAalpha2-3Galbeta1 4(Fucalpha1-3)(6-sulfo)GlcNAcbeta1-3Galbeta1-3(SAalpha2-3Galbeta1-4GlcNAcbeta1 6)GalNAc. However, if sulfo sLex glycans are supplemented with separate sulfated, nonfucosylated O-glycans, saccharides in O-6, O-8, or O-9, putatively carrying MECA-79 epitopes, could form multiglycan binding epitopes for L-selectin. PMID- 11895803 TI - Increased soluble and platelet-associated CD40 ligand in essential thrombocythemia and reactive thrombocytosis. AB - CD40 ligand (CD40L) is expressed on activated CD4(+) T lymphocytes and at the activated platelet surface. A circulating soluble form of CD40L (sCD40L) is generated by the way of a proteolytic cleavage. We measured sCD40L in the plasma of either healthy subjects; patients with inflammatory disorders and low, normal, or high platelet count (reactive thrombocytosis); or patients with essential thrombocythemia (ET). A tight correlation was found between the platelet count and plasma sCD40L. ET patients had the highest levels of sCD40L. Platelet associated CD40L was increased in ET and reactive thrombocytosis, conditions associated with increased platelet regeneration. Platelet-associated CD40L was released upon platelet activation. In conclusion, platelets appear as a reservoir of CD40L that may be a major contributor to circulating sCD40L. Platelet associated CD40L may be a potential marker of platelet regeneration. PMID- 11895804 TI - Transient hematologic and clinical effect of E21R in a child with end-stage juvenile myelomonocytic leukemia. AB - E21R is a modified granulocyte macrophage-colony-stimulating factor (GM-CSF) protein which results in antagonism of GM-CSF function via selective binding to the GM-CSF receptor complex. Juvenile chronic myelomonocytic leukemia (JMML) is a rare leukemia where spontaneous proliferation of myeloid and monocytic precursors in patients' bone marrow cultures is dependent on GM-CSF. For patients who progress after systemic chemotherapy, there are no effective therapies. In vitro and in vivo studies in an animal model demonstrating that E21R exerts an antileukemic action prompted us to consider its potential utility in a child with end-stage JMML. E21R was well-tolerated during the 3 courses of subcutaneous treatment. A clear in vivo efficacy was observed after 2 courses of E21R but the disease appeared completely refractory during the third course. This novel therapeutic approach clearly deserves further evaluation in JMML. PMID- 11895805 TI - Expression of cyclic adenosine monophosphate response-element binding protein in acute leukemia. AB - Cyclic adenosine monophosphate response-element binding protein (CREB) is a nuclear protein that regulates expression of genes that control cell proliferation, differentiation, and survival. To analyze CREB expression in leukemia cells, we conducted Western blot analysis of bone marrow cells obtained from patients with acute lymphoblastic leukemia, patients with acute myeloid leukemia, and patients without active leukemia. CREB was expressed at a higher frequency in bone marrow cells from patients with acute lymphoid or myeloid leukemia than in patients with leukemia remission or without leukemia. Our results indicate that CREB expression could be a useful marker for leukemia in patients with acute disease and suggest a role for CREB in leukemogenesis. PMID- 11895806 TI - p16(INK4a) immunocytochemical analysis is an independent prognostic factor in childhood acute lymphoblastic leukemia. AB - We investigated the prognostic value of p16(INK4a) immunocytochemistry (ICC) analysis in 126 cases of newly diagnosed childhood acute lymphoblastic leukemia (ALL). The incidence of negative p16(INK4a) ICC was 38.1% and was more frequent in T-lineage ALL. Overall survival (OS) and event-free survival (EFS) were significantly higher in patients with positive p16(INK4a) ICC than in patients with negative ICC (6 years OS, 90% versus 63%, P =.0014; 6 years EFS, 77.8% versus 55%, P =.0033). The p16(INK4a) ICC remained a significant prognostic factor within the subgroup of B-precursor ALL. Multivariate analysis showed that negative p16(INK4a) ICC was an independent prognostic factor for OS (relative risk [RR], 3.38; P =.02) and EFS (RR, 2.49; P =.018). Sequential study showed that p16(INK4a) expression remained stable during first relapse in most patients. These findings indicate that p16(INK4a) ICC is an independent factor of outcome in childhood ALL. PMID- 11895807 TI - Shame: the elephant in the room. PMID- 11895808 TI - Declining altruism in medicine. PMID- 11895809 TI - Epidemic of cardiovascular disease in South Asians. PMID- 11895810 TI - Time to abandon the "tendinitis" myth. PMID- 11895811 TI - Deprofessionalising doctors? PMID- 11895814 TI - Public less worried about MMR vaccine than many other issues. PMID- 11895812 TI - Woman makes legal history in right to die case. PMID- 11895815 TI - Improved anthrax vaccine is needed, claims report. PMID- 11895816 TI - Children at risk after sperm donor develops late onset genetic disease. PMID- 11895819 TI - Woman may face death penalty in postnatal depression case. PMID- 11895820 TI - Early evidence of ethnic differences in cardiovascular risk: cross sectional comparison of British South Asian and white children. AB - OBJECTIVES: To examine whether British South Asian children differ in insulin resistance, adiposity, and cardiovascular risk profile from white children. DESIGN: Cross sectional study. SETTING: Primary schools in 10 British towns. PARTICIPANTS: British children aged 8 to 11 years (227 South Asian and 3415 white); 73 South Asian and 1287 white children aged 10 and 11 years provided blood samples (half fasting, half after glucose load). MAIN OUTCOME MEASURES: Insulin concentrations, anthropometric measures, established cardiovascular risk factors. RESULTS: Mean ponderal index was lower in South Asian children than in white children (mean difference -0.43 kg/m(3), 95% confidence interval -0.13 kg/m(3) to -0.73 kg/m(3)). Mean waist circumferences and waist:hip ratios were similar. Mean insulin concentrations were higher in South Asian children (percentage difference was 53%, 14% to 106%, after fasting and 54%, 19% to 99%, after glucose load), though glucose concentrations were similar. Mean heart rate and triglyceride and fibrinogen concentrations were higher among South Asian children; serum total, low density lipoprotein, and high density lipoprotein cholesterol concentrations were similar in the two groups. Differences in insulin concentrations remained after adjustment for adiposity and other potential confounders. However, the relations between adiposity and insulin concentrations (particularly fasting insulin) were much stronger among South Asian children than among white children. CONCLUSIONS: The tendency to insulin resistance observed in British South Asian adults is apparent in children, in whom it may reflect an increased sensitivity to adiposity. Action to prevent non-insulin dependent diabetes in South Asian adults may need to begin during childhood. PMID- 11895821 TI - Qualitative study of evidence based leaflets in maternity care. AB - OBJECTIVE: To examine the use of evidence based leaflets on informed choice in maternity services. DESIGN: Non-participant observation of 886 antenatal consultations. 383 in depth interviews with women using maternity services and health professionals providing antenatal care. SETTING: Women's homes; antenatal and ultrasound clinics in 13 maternity units in Wales. PARTICIPANTS: Childbearing women and health professionals who provide antenatal care. INTERVENTION: Provision of 10 pairs of Informed Choice leaflets for service users and staff and a training session in their use. MAIN OUTCOME MEASURES: Participants' views and commonly observed responses during consultations and interviews. RESULTS: Health professionals were positive about the leaflets and their potential to assist women in making informed choices, but competing demands within the clinical environment undermined their effective use. Time pressures limited discussion, and choice was often not available in practice. A widespread belief that technological intervention would be viewed positively in the event of litigation reinforced notions of "right" and "wrong" choices rather than "informed" choices. Hierarchical power structures resulted in obstetricians defining the norms of clinical practice and hence which choices were possible. Women's trust in health professionals ensured their compliance with professionally defined choices, and only rarely were they observed asking questions or making alternative requests. Midwives rarely discussed the contents of the leaflets or distinguished them from other literature related to pregnancy. The visibility and potential of the leaflets as evidence based decision aids was thus greatly reduced. CONCLUSIONS: The way in which the leaflets were disseminated affected promotion of informed choice in maternity care. The culture into which the leaflets were introduced supported existing normative patterns of care and this ensured informed compliance rather than informed choice. PMID- 11895822 TI - Use of evidence based leaflets to promote informed choice in maternity care: randomised controlled trial in everyday practice. AB - OBJECTIVE: To assess the effect of leaflets on promoting informed choice in women using maternity services. DESIGN: Cluster trial, with maternity units randomised to use leaflets (intervention units) or offer usual care (control units). Data collected through postal questionnaires. SETTING: 13 maternity units in Wales. PARTICIPANTS: Four separate samples of women using maternity services. Antenatal samples: women reaching 28 weeks' gestation before (n=1386) and after (n=1778) the intervention. Postnatal samples: women at eight weeks after delivery before (n=1741) and after (n=1547) the intervention. INTERVENTION: Provision of 10 pairs of Informed Choice leaflets for service users and midwives and a training session for staff in their use. MAIN OUTCOME MEASURES: Change in the proportion of women who reported exercising informed choice. SECONDARY OUTCOMES: changes in women's knowledge; satisfaction with information, choice, and discussion; and possible consequences of informed choice. RESULTS: There was no change in the proportion of women who reported that they exercised informed choice in the intervention units compared with the control units for either antenatal or postnatal women. There was a small increase in satisfaction with information in the antenatal samples in the intervention units compared with the control units (odds ratio 1.40, 95% confidence interval 1.05 to 1.88). Only three quarters of women in the intervention units reported being given at least one of the leaflets, indicating problems with the implementation of the intervention. CONCLUSION: In everyday practice, evidence based leaflets were not effective in promoting informed choice in women using maternity services. PMID- 11895823 TI - Trends in demand for emergency ambulance services in Wiltshire over nine years: observational study. PMID- 11895824 TI - After Bristol: putting patients at the centre. PMID- 11895825 TI - Cost effectiveness of continuing professional development in health care: a critical review of the evidence. PMID- 11895827 TI - Abdominal pain in acute infectious mononucleosis. PMID- 11895826 TI - Recent developments in neurology. PMID- 11895828 TI - ABC of clinical electrocardiography. Junctional tachycardias. PMID- 11895829 TI - What action should be taken to prevent spread of vancomycin resistant enterococci in European hospitals? PMID- 11895830 TI - Designing studies to ensure that estimates of test accuracy are transferable. PMID- 11895831 TI - The "redisorganisation" of the NHS. Is mass dysmorphophobia a better term? PMID- 11895832 TI - Effectiveness of guidelines on persistent glue ear in children. Authors' estimates of size of impact are probably excessive. PMID- 11895833 TI - Why general practitioners do not implement evidence. Evidence seems to change frequently. PMID- 11895834 TI - Adult obesity and growth in childhood. Fuel mediated teratogenesis driven by maternal obesity may be responsible for pandemic of obesity. PMID- 11895835 TI - Implications of childhood obesity for adult health. Message on childhood obesity was missed. PMID- 11895836 TI - Combining high quality clinical and interpersonal care. PMID- 11895837 TI - Social deprivation affects outcome of nocturnal enuresis. PMID- 11895839 TI - Price of interferon beta is similar in UK and Australia. PMID- 11895838 TI - Breast screening seems driven by belief rather than evidence. PMID- 11895840 TI - Data on take home naloxone are unclear but not condemnatory. PMID- 11895841 TI - Venous ulcers may be associated with gravitational eczema. PMID- 11895842 TI - Violence may be serious in men with body dysmorphic disorder. PMID- 11895843 TI - Chinese medicines for slimming still cause health problems. PMID- 11895844 TI - Pharmaceutical packaging can induce confusion. PMID- 11895851 TI - Skilled use of DNA polymorphisms as a tool for polygenic cancers. AB - Association studies are assumed to be an efficient method of deciding whether a gene or its variant is important for cancer. Sequencing data on 30 000 human genes suggest that an average gene contains one to two single nucleotide polymorphisms (SNP), and high through-put technologies have become available for fast genotyping. Because no functional data are available for most SNPs, the result of the large-scale genotyping effort will be a huge amount of data of unknown biological significance. We discuss here the approaches in study design and reporting that will reduce the spread of false positive data and optimize scientific progress in the genotyping field. PMID- 11895852 TI - Gene-environment interaction and aetiology of cancer: what does it mean and how can we measure it? AB - One form of defence against cancer development involves a series of genes whose role is to metabolize and excrete potentially toxic compounds and to repair subtle mistakes in DNA. Much laboratory and epidemiological research over the past decade has concentrated on the identification of these genes and an assessment of their role in cancer aetiology. Of particular interest has been whether the risk of cancer associated with a particular environmental exposure differs with respect to functionally different polymorphisms of these genes, i.e. gene-environment interaction. A large number of studies have been conducted for numerous genes and also for all common cancer sites, although results have been very inconsistent and therefore inconclusive. This is partially due to the inadequate sample size of most studies to detect modest effects and the over reporting of positive associations identified in subgroups of the dataset. There is also much confusion about the meaning of "gene-environment interaction", what type of studies should be conducted to study it and also how it should be measured. Furthermore, the very purpose of these studies is not clear; are they attempting to identify high-risk individuals, or are they simply trying to further understand the cancer process? This review will explore these questions and provide some recommendations to help ensure that the next phase of gene environment interaction studies, which are likely to be much larger and based on many more genes, also provide some clearer answers. PMID- 11895853 TI - Flow cytometric analysis of the cell cycle phase specificity of DNA damage induced by radiation, hydrogen peroxide and doxorubicin. AB - We have optimized a flow cytometric DNA alkaline unwinding assay to increase the sensitivity in detecting low levels of DNA damage (strand breaks and alkali labile sites) and to permit the measurement of the extent of DNA damage within each cell cycle compartment. The lowest gamma radiation dose that induced detectable DNA damage in each cell cycle phase of HeLa and CEM cells was 10 cGy. The lowest H(2)O(2) concentration that induced detectable DNA damage in each cell cycle phase was 0.5 microM in HeLa cells, and 1-2.5 TmicroM in CEM cells. For both HeLa cells and CEM cells, DNA damage in each cell cycle compartment increased approximately linearly with increasing doses of gamma radiation and H(2)O(2). Although untreated HeLa and CEM cells in S phase consistently exhibited greater DNA unwinding than did G(1) or G(2) cells (presumably due to DNA strand breaks associated with replication forks), there was no difference between the susceptibility of G(0)/G(1), S and G(2)/M phase cells to DNA damage induced by gamma radiation or H(2)O(2), or in the rate of repair of this damage. In each cell cycle phase, the susceptibility to gamma radiation-induced DNA damage was greater in CEM cells than in HeLa cells. In contrast to the lack of cell cycle phase-specific DNA damage induced by exposure to gamma radiation or H(2)O(2), the cancer chemotherapeutic drug doxorubicin (adriamycin) predominantly induced DNA damage in G(2) phase cells. PMID- 11895854 TI - Human cells deficient in p53 regulated p21(waf1/cip1) expression exhibit normal nucleotide excision repair of UV-induced DNA damage. AB - Cancer development requires the accumulation of numerous genetic changes, which are believed to initiate through the presence of unrepaired lesions in the genome. In the absence of proficient repair, genotoxic agents can lead to crucial mutations of vital cellular genes via replication of damaged DNA. Many cell cycle regulatory proteins are known to modulate the repair capacity and consequently the fate of cells. We and others have recently shown that p53 tumor suppressor gene product is required for efficient global genomic repair (GGR) but not the transcription coupled repair (TCR) of the nucleotide excision repair (NER) sub pathways. In order to discern the nature of the p53 modulation to be direct or indirect through a downstream mediator, we have investigated the processing of UV radiation induced lesions in human colon carcinoma, HCT116 cells expressing wild type p53 but having different p21(waf1cip1) (hereafter p21) genotypes (p21+/+, p21+/-, p21-/-). Following 20 J/m(2) UV, all the three cell lines showed rapid increase in p53 protein but the accompanying increase in the expression of its downstream target protein p21 could only be seen in p21+/+ and p21+/- cells and not in p21-/- cells. Nevertheless, an absence of detectable p21 protein in deficient cells had no demonstrable effect on DNA repair response to UV irradiation, as measured by an immunoassay to detect removal of UV photoproducts from genomic DNA (GGR) and by individual strand specific removal of endonuclease sensitive CPD from a target gene fragment (TCR). Introduction of cytomegalovirus (CMV)-driven luciferase reporter plasmid, UV damaged in vitro, into the un irradiated cells of varying p21 background, revealed a relatively small but statistically significant decrease in the reporter expression in the host p21-/- as compared with p21+/+ and p21+/- HCT116 cells. Super-expression of p21 protein upon reintroduction of p21 expression construct, showed an enhanced recovery of UV damaged reporter activity that was not greatly different from a similar enhancement observed with undamaged plasmid reporter DNA. Taken together, the results indicate that (i) the p21 protein does not have a significant role in the repair of genomic DNA at chromosomal level; (ii) the well-established p53 dependent modulation of NER is distinct and independent of its cell cycle checkpoint function; and (iii) the reproducible enhancing effect of p21 expression observed through host cell reactivation (HCR) of extrachromosomal DNA is mainly attributable to an effect exerted on transcription rather than repair. PMID- 11895855 TI - Quantitative trait locus mapping of susceptibilities to butylated hydroxytoluene induced lung tumor promotion and pulmonary inflammation in CXB mice. AB - We have reported previously [Bauer,A.K. et al. (2001) Exp. Lung Res., 27, 197 216] that the 13 CXB recombinant inbred mouse strains derived from BALB/cByJ and C57BL/6J progenitors vary in their responsiveness to both lung tumor promotion and pulmonary inflammation induced by chronic administration of butylated hydroxytoluene (BHT). Herein we have applied these data, along with markers known to be polymorphic among these strains, to conduct linkage analysis of these susceptibilities. This enabled us to assign provisional quantitative trait loci (QTL) that govern these strain variations in susceptibility as a genetic approach to assessing the influence of inflammation on tumorigenesis. A Chr 15 (39.1-55.6 cM) QTL regulated susceptibility to two-stage carcinogenesis, a protocol in which chronic BHT exposure followed a single urethane injection; a similar QTL on Chr 15 (46.7-61.7 cM) influenced BHT induction of cyclooxygenase-2 (COX-2) expression. A Chr 18 (37-41 cM) QTL modulated both the number of lung tumors induced by 3-methylcholanthrene (MCA) injection with subsequent treatment with BHT as well as BHT-induced ingress of macrophages into airways. Other chromosomal sites that affected either the degree of BHT-elicited macrophage infiltration, Chr 9 (48-61 cM), or COX-2 induction, Chr 10 (59-65 cM), were reported to influence susceptibility to lung tumorigenesis in other strains. The fact that common chromosomal locations regulate both inflammation and carcinogenesis suggests a pathogenic role of inflammatory mediators in tumor development that may be exploited for chemoprevention of lung cancer. PMID- 11895857 TI - Resveratrol enhances the expression of non-steroidal anti-inflammatory drug activated gene (NAG-1) by increasing the expression of p53. AB - Dietary phenolic substances including resveratrol, a stillbene compound, are found in several fruits and vegetables, and these compounds have been reported to have anti-oxidant, anti-inflammatory and antitumorigenic activities. However, the molecular mechanisms underlying the antitumorigenic or chemopreventive activities of these compounds remain largely unknown. The expression of NAG-1 [non-steroidal anti-inflammatory (NSAID) drug-activated gene-1], a member of the transforming growth factor-beta (TGF-beta) superfamily, has been shown to be associated with pro-apoptotic and antitumorigenic activities. Here, we have demonstrated that resveratrol induces NAG-1 expression and apoptosis in a concentration-dependent manner. Resveratrol increases the expression of p53, tumor suppressor protein, prior to NAG-1 induction, indicating that NAG-1 expression by resveratrol is mediated by p53 expression. We also show that the p53 binding sites within the promoter region of NAG-1 play a pivotal role to control NAG-1 expression by resveratrol. Derivatives of resveratrol were examined for NAG-1 induction, and the data suggest that resveratrol-induced NAG-1 and p53 induction is not dependent on its anti-oxidant activity. The data may provide linkage between p53, NAG-1 and resveratrol, and in part, a new clue to the molecular mechanism of the antitumorigenic activity of natural polyphenolic compounds. PMID- 11895856 TI - CagA status of Helicobacter pylori infection and p53 gene mutations in gastric adenocarcinoma. AB - Infection with Helicobacter pylori (H. pylori) increases stomach cancer risk. Helicobacter pylori strains with the cag pathogenicity island (PAI) induce more severe inflammation in the gastric epithelium and are more strongly associated with stomach cancer risk than strains lacking the PAI. We examined whether the prevalence of somatic p53 mutation in gastric adenocarcinoma differed between subjects with and without infection with CagA(+) (a marker for the PAI) H. pylori strains. DNA from 105 microdissected tumor specimens was analyzed for mutation in exons 5-8 of the p53 gene by polymerase chain reaction-based single-strand conformation polymorphism followed by direct DNA sequencing. Enzyme-linked immunosorbent assays for IgG antibodies against H. pylori and CagA were performed on sera collected 2-31 years prior to cancer diagnosis. Tumors from CagA(+) subjects were significantly more likely to have p53 mutations than tumors from CagA(-) subjects (including H. pylori- and H. pylori(+)/CagA(-)): odds ratio = 3.72; 95% confidence interval, 1.06-13.07 after adjustment for histologic type and anatomic subsite of tumor and age at diagnosis and sex of subjects. Mutations were predominantly insertions and deletions (43%) as well as transition mutations at CpG dinucleotides (33%). The data suggest that CagA(+) H. pylori infection, when compared with CagA(-) infection or the absence of H. pylori infection, is associated with a higher prevalence of p53 mutation in gastric adenocarcinoma. PMID- 11895858 TI - Generation and characterization of p53 null transformed hepatic progenitor cells: oval cells give rise to hepatocellular carcinoma. AB - Oval cells are bipotential liver stem cells able to differentiate into hepatocytes and bile duct epithelia. In normal adult liver oval cells are quiescent, existing in low numbers around the periportal region, and proliferate following severe, prolonged liver trauma. There is evidence implicating oval cells in the development of hepatocellular carcinoma, and hence the availability of an immortalized oval cell line would be invaluable for the study of liver cell lineage differentiation and carcinogenesis. A novel approach in the generation of cell lines is the use of the p53 knockout mouse. Absence of p53 allows a cell to cycle past the normal Hayflick limit, rendering it immortalized, although subsequent genetic alterations are thought necessary for transformation. p53 knockout mice were fed a choline-deficient, ethionine-supplemented diet, previously shown to increase oval cell numbers in wild-type mice. The oval cells were isolated by centrifugal elutriation and maintained in culture. Colonies of hepatic cells were isolated and characterized with respect to phenotype, growth characteristics and tumorigenicity. Analysis of gene expression by Northern blotting and immunocytochemistry suggests they are oval-like cells by virtue of albumin and transferrin expression, as well as the oval cell markers alpha fetoprotein, M(2)-pyruvate kinase and A6. Injection into athymic nude mice shows the cell lines are capable of forming tumors which phenotypically resemble hepatocellular carcinoma. Thus, the use of p53 null hepatic cells successfully generated immortalized and tumorigenic hepatic stem cell lines. The results presented support the idea that deleting p53 allows immortalization and contributes to the transformation of the oval-like cell lines. Further, the tumorigenic status of the cell lines is direct evidence for the participation of oval cells in the formation of hepatocellular carcinoma. PMID- 11895859 TI - Apoptosis of squamous cells at different stages of carcinogenesis following 4-HPR treatment. AB - Squamous cell carcinoma (SCC) is the end product of a multistep process characterized by a progression from normal epithelial cells through metaplastic or dysplastic intraepithelial changes that evolve into invasive cancer. Since retinamides have shown promising in vivo anti-tumoral activity, we studied effects and effector mechanisms of the synthetic retinoid N-(4 hydroxyphenyl)retinamide (4-HPR) on squamous cells at progressing stages of tumorigenesis. To this end, an in vitro model of squamous carcinogenesis consisting of normal human keratinocytes, human papilloma virus (HPV) immortalized keratinocytes (UP) and tumorigenic HPV-immortalized/v-Ha-ras transfected keratinocytes (UPR) was used. 4-HPR treatment affected cell growth at doses higher than 1.5 microM. Flow cytometric measurements of DNA content and annexin V revealed that cell growth decrease was mainly due to apoptosis at 4-HPR concentrations of or below 15 microM, and necrosis at higher concentrations. The effects were similar in the three cell types of the in vitro model, as well as in three SCC cell lines, suggesting that sensitivity to 4-HPR is independent of the degree of squamous cell tumorigenesis in the in vitro model. We further investigated whether mitochondrial damage was involved in the course of 4-HPR induced apoptosis. Treatment of squamous cells with the antioxidant L-ascorbic acid inhibited apoptosis, indicating that 4-HPR increases production of free radicals. Measures of mitochondrial membrane potentials showed that 4-HPR induced membrane permeability transition (MPT), and that MPT-inhibitors were able to reduce apoptosis. This indicates that MPT is involved in apoptosis signalling by 4-HPR. Finally, we studied the role of caspases. We found that caspases 8, 9 and 3 participate in 4-HPR-mediated apoptosis of squamous cells, and that MPT is an upstream event that regulates caspase activity. Caspase 8 was activated independently of the Fas-Fas ligand pathway. PMID- 11895860 TI - Nrf2 transactivator-independent GSTP1-1 expression in "GSTP1-1 positive" single cells inducible in female mouse liver by DEN: a preneoplastic character of possible initiated cells. AB - Whether single cells immunohistochemically positive for glutathione S-transferase P1-1 (GSTP1-1) induced in the female mouse liver by DEN (Hatayama et al., Carcinogenesis, 14, 537-538, 1993) are precursor initiated cells of preneoplastic foci, is of importance in chemical hepatocarcinogenesis. Nrf2 transactivates a wide variety of ARE (anti-oxidant response element)-mediated enzymes including GSTP1-1. Quantitative examination revealed that the basal expression of hepatic GSTP1-1 was 60% lower in Nrf2 gene knock-out female mice(-/-) than in wild type females, and that treatment with butyrated hydroxyanisole (BHA) increased by 10 fold GSTP1-1 expression in the liver of wild type female mice but not in knockout female mice(-/-). Despite the lack of Nrf2, GSTP1-1-positive single cells were detected in livers of DEN-treated female(-/-) 3 months after treatment. Subsequent BHA feeding to the positive cell-bearing females for one more week clearly showed that the single cells were detectable with females(-/-) but not with females(+/+,+/-) due to the strong induction of GSTP1-1 in the surrounding hepatocytes. The sensitivity to DEN hepatocarcinogenesis was not significantly different among genotypes. These results demonstrate that Nrf2 is regulatory in normal hepatocytes but not in the single cells positive for GSTP1-1 inducible in the female mouse liver by DEN. The transcriptional distinction observed for the DEN-transformants is suggestive of a preneoplastic character of precursor initiated cells. PMID- 11895861 TI - Effects of genetic background on prostate and taste bud carcinogenesis due to SV40 T antigen expression under probasin gene promoter control. AB - The incidence of prostate carcinomas in African-American men is greater than in white men, indicating genetic factors are involved in risk of this neoplasia. Recently, we have developed a transgenic rat model of prostate cancer, featuring development of malignancies within 15 weeks of age at very high incidence. Male transgenic rats with a Sprague-Dawley genetic background were mated with wild type females of F344, Wistar and ACI strains. F1 male transgenic hybrids with female Wistar and ACI rats had significantly lowered incidences of prostate carcinomas. However, the serum level of testosterone, and expression of the transgene, probasin, and the androgen receptor did not correlate with the strain variation in tumor development. Furthermore, immunohistochemical analysis of the SV40 Tag and the androgen receptor also did not reveal any differences between the strains. The transgenic rats additionally developed taste bud neuroblastomas at 100% incidence and this was suppressed in F1 male transgenic offspring with the ACI, but not the other strains. These results clearly show that genetic background influences prostate carcinogenesis and taste bud tumorigenesis in rats and that the present transgenic rats could provide a good model to identify specific factors. PMID- 11895862 TI - Influence of nitric oxide on the generation and repair of oxidative DNA damage in mammalian cells. AB - We have analysed the effects of endogenously and exogenously generated nitric oxide (NO) in cultured mammalian fibroblasts on: (i) the steady-state (background) levels of oxidative DNA base modifications; (ii) the susceptibility of the cells to the induction of additional DNA damage and micronuclei by H(2)O(2); and (iii) the repair kinetics of various types of DNA modifications. Steady-state levels of oxidative DNA base modifications, measured by means of an alkaline elution assay in combination with the repair endonuclease Fpg protein, were similar in NO-overproducing B6 mouse fibroblasts stably transfected with an inducible NO synthase (iNOS) and in control cells. Increased oxidative damage was only observed after exposure to high (toxic) concentrations of exogenous NO generated by decomposition of dipropylenetriamine-NONOate (DPTA-NONOate). Under these conditions, the spectrum of DNA modifications was similar to that induced by 3-morpholinosydnonimine, which generates peroxynitrite. The repair rate of additional oxidative DNA base modifications induced by photosensitization was not affected by the endogenous NO generation in the iNOS-transfected cells. However, it was completely blocked after pre-treatment with DPTA-NONOate at concentrations that did not cause oxidative DNA damage by themselves. In contrast, the repair of DNA single-strand breaks, sites of base loss (AP sites) and UVB-induced pyrimidine photodimers, was not affected. The endogenous generation of NO in the iNOS-transfected fibroblasts was associated with a protection from DNA single strand break formation and micronuclei induction by H(2)O(2). These results indicate that NO generates cellular DNA damage only inefficiently and can even protect from DNA damage by H(2)O(2), but it selectively inhibits the repair of oxidative DNA base modifications. PMID- 11895863 TI - Effects of dairy products on heterocyclic aromatic amine-induced rat colon carcinogenesis. AB - Heterocyclic aromatic amines (HAA) are initiating agents of colon carcinogenesis in animals and are suspected in the aetiology of human colon cancer. In the context of prevention, it seems interesting to test possible protective compounds, such as fermented milk, against HAA food carcinogens. Male F344 rats were used in a model of HAA-induced colon carcinogenesis. The HAA, 2-amino-3 methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (ratio 1:1:1) were administered in food for a 7 week induction period, with a cumulative dose of 250 mg of the HAA, per kg body weight. Four different diets were given to four rat groups: supplemented with 20% water, 30% non-fermented milk, 30% Bifidobacterium animalis DN-173 010 fermented milk and 30% Streptococcus thermophilus DN-001 158 fermented milk. Fecal mutagenicity was quantified during the induction period. At the end of the treatment, DNA lesion levels were determined in the liver and colon using the number of 8-oxo-7,8-dihydro 2'desoxyguanosine (8-oxodGuo) oxidized bases, "3D Test" and comet assay. The metabolic activity of hepatic and colon cytochrome P450 (CYP450) 1A1 and 1A2 was also evaluated. Aberrant colon crypts were scored, 8 weeks after the last HAA treatment. The results showed that dairy products decreased the incidence of aberrant crypts in rats: 66% inhibition with the milk-supplemented diet, 96% inhibition with the B.animalis fermented milk-supplemented diet and 93% inhibition with the S.thermophilus fermented milk-supplemented diet. Intermediate biomarkers showed that there was a decrease in HAA metabolism, fecal mutagenicity and colon DNA lesions. These results demonstrate the early protective effect of milk in the carcinogenesis process. This effect being more pronounced in the case of milk fermented by lactic acid bacteria. PMID- 11895864 TI - Immunohistochemical detection of a substituted 1,N(2)-ethenodeoxyguanosine adduct by omega-6 polyunsaturated fatty acid hydroperoxides in the liver of rats fed a choline-deficient, L-amino acid-defined diet. AB - Endogenous lipid peroxidation products react with DNA and form exocyclic DNA adducts. The purpose of this study was to investigate the in vivo formation of 7 (2-oxo-heptyl)-substituted 1,N(2)-etheno-2'-deoxyguanosine adduct (Oxo-heptyl varepsilondG), one of the major products from the reaction of 13 hydroperoxyoctadecadienoic acid (13-HPODE) with dG. The monoclonal antibody specific to Oxo-heptyl-varepsilondG was prepared using a chemically synthesized conjugate of Oxo-heptyl-varepsilondG and carrier protein as immunogen. The characterization showed that the obtained antibody (mAb6A3) is specific to the Oxo-heptyl-varepsilondG moiety. Using the novel antibody, the presence of the Oxo heptyl-varepsilondG adduct in vivo was immunohistochemically revealed in the liver of rats fed a choline-deficient, L-amino acid-defined diet, an endogenous carcinogenesis model, for 3 days. In addition, the Oxo-heptyl-varepsilondG formation was confirmed in DNAs treated with peroxidized linoleic acid, arachidonic acid and gamma-linolenic acid, respectively, suggesting that the hydroperoxides of omega-6 polyunsaturated fatty acids could be the potential sources of Oxo-heptyl-varepsilondG formation in vivo. Collectively, the results in this study suggest the first evidence that the formation of Oxo-heptyl varepsilondG, the omega-6 lipid hydroperoxide-mediated DNA adduct, may be a potential biomarker for the early phase of carcinogenesis. PMID- 11895865 TI - Change in the gene expression of hepatic tamoxifen-metabolizing enzymes during the process of tamoxifen-induced hepatocarcinogenesis in female rats. AB - Altered gene expression of the enzymes responsible for tamoxifen metabolism during the process of tamoxifen-induced hepatocarcinogenesis in female Sprague Dawley rats was examined by the RT-PCR method. Treatment of rats with tamoxifen (20 mg/kg body/day) for 52 weeks, but not the 1 day, 2 or 12 week treatments, resulted in the formation of the liver hyperplastic nodules. The gene expression of CYP3A subfamily enzymes, especially CYP3A1, responsible for not only detoxification (N-demethylation) but also activation (alpha-hydroxylation) of tamoxifen, was increased by the tamoxifen treatments for 2 and 12 weeks, whereas after the 52 week treatment, the expression in the induced nodules returned to the control level. The gene expression of SULT2A subfamily sulfotransferases, especially HSTa, responsible for metabolic activation of alpha-hydroxytamoxifen was decreased to a level <20% of the control in the nodules, although no significant change in the expression was observed in the liver of rats treated with tamoxifen for 1 day, 2 or 12 weeks. On the other hand, the gene expression of CYP3A2 and flavin-containing monooxygenase 1 (FMO1), responsible for the N demethylation and N-oxidation, respectively, of tamoxifen was increased in a time dependent fashion up to the 52 week treatment. Although the gene expression of UDP-glucuronosyltransferase(s), which might be responsible for detoxification of tamoxifen, was also increased by the tamoxifen treatment for 2 or 12 weeks, it decreased to the control level in the nodules after the 52 week treatment. The present findings demonstrate that in the early stage of the formation of the liver hyperplastic nodules by tamoxifen, the genes of the enzymes responsible for not only detoxification but also activation of tamoxifen were activated, whereas in the later stage (in the nodules), the genes of the detoxification enzymes, CYP3A2 and FMO1, remained active, but those of the activation enzymes such as CYP3A1 and HSTa were suppressed. PMID- 11895866 TI - Silymarin inhibits growth and causes regression of established skin tumors in SENCAR mice via modulation of mitogen-activated protein kinases and induction of apoptosis. AB - This study reports in vivo therapeutic efficacy of silymarin against skin tumors with mechanistic rationale. 7,12-Dimethylbenz[a]anthracene-12-O-tetradecanoyl phorbol-13-acetate (DMBA-TPA)-induced established skin papilloma (tumor)-bearing SENCAR mice were fed with 0.5% silymarin in AIN-93M-purified diet (w/w), and both tumor growth and regression were monitored during 5 weeks of feeding regimen. Silymarin feeding significantly inhibited (74%, P < 0.01) tumor growth and also caused regression (43%, P < 0.01) of established tumors. Proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling immunohistochemical staining of tumors showed that silymarin decreases proliferation index by 48% (P < 0.001) and increases apoptotic index by 2.5-fold (P < 0.001), respectively. Skin tumor growth inhibition and regression by silymarin were also accompanied by a strong decrease (P < 0.001) in phospho ERK1/2 levels in tumors from silymarin-fed mice compared with controls. In the studies evaluating bioavailability and physiologically achievable level of silymarin (as silibinin) in plasma, skin tumor, skin, liver, lung, mammary gland and spleen, we found 10, 6.5, 3.1, 13.7, 7.7, 5.9 and 4.4 microg silibinin/ml plasma or per gram tissue, respectively. In an attempt to translate these findings to human skin cancer and to establish biological significance of physiologically achievable level, effect of plasma concentration of silibinin was next examined in human epidermoid carcinoma A431 cells. Silibinin treatment of cells in culture at 12.5, 25 (plasma level) and 50 microM doses resulted in 30 74% (P < 0.01-0.001) growth inhibition and 7-42% death of A431 cells in a dose- and time-dependent manner; apoptosis was identified as a cell death response by silibinin. Similar silibinin treatments also resulted in a significant decrease in phospho-mitogen-activated protein kinase/extracellular signal-regulated protein kinase 1/2 (MAPK/ERK1/2) levels, but an up-regulation of stress-activated protein kinase/jun NH(2)-terminal kinase (SAPK/JNK1/2) and p38 mitogen-activated protein kinase (p38 MAPK) activation in A431 cells. The use of MEK1 inhibitor, PD98059, showed that inhibition of ERK1/2 signaling, in part, contributes to silibinin-caused cell growth inhibition. Together, the data suggest that an inhibition of ERK1/2 activation and an increased activation of JNK1/2 and p38 by silibinin could be possible underlying molecular events involved in inhibition of proliferation and induction of apoptosis in A431 cells. These data suggest that silymarin and/or its major active constituent silibinin could be an effective agent for both prevention and intervention of human skin cancer. PMID- 11895867 TI - Lung tumors in A/J mice exposed to environmental tobacco smoke: estimated potency and implied human risk. AB - Directly inhaled tobacco smoke is a recognized human lung carcinogen, and epidemiological studies suggest relative risks of about 1.2-1.4 for nonsmoking spouses of smokers typically exposed to environmental tobacco smoke (ETS). While many individual ETS components have been shown experimentally to induce lung tumors, ETS itself was only recently shown to induce lung tumors in a series of studies in which strain A/J mice were exposed to well-defined ETS atmospheres. Data from these studies indicate that ETS exposure clearly can increase combined malignant and benign lung tumors in multiple experiments involving male and female A/J mice, and thus provide convincing evidence that ETS is a positive mouse carcinogen. Tumorigenic potencies estimated from these A/J mouse bioassay data predict a corresponding range of increased human risk (0.2-0.5%) that overlaps that implied by case-control studies showing increased lung cancer risks in lifelong nonsmokers married to smokers. In A/J mice exposed to a significantly tumorigenic ETS concentration, lung tumors were found to be significantly smaller than those in corresponding control mice, and mice so exposed for 9 months had significantly fewer tumors/animal than mice exposed for 5 months followed by 4 months in filtered ETS-free air. These findings support hypotheses that ETS does not promote growth of spontaneous neoplastic foci in A/J mice, and that ETS induced lung-tumor risk in A/J mice occurs predominantly by genotoxic effects that can be suppressed by reduced cell proliferation associated with chronic, high-level ETS exposure. The results obtained add to evidence that A/J mouse lung tumors induced by ETS provide a relevant biological model of ETS-induced human lung tumors. PMID- 11895868 TI - Mechanism of lovastatin-induced apoptosis in intestinal epithelial cells. AB - We earlier showed that lovastatin potentiated the chemopreventive effects of sulindac against colon neoplasia in a rodent model and augments apoptosis induced by 5-FU and cisplatin in human colon cancer cells. In the present study, we investigated effects of lovastatin in spontaneously immortalized rat intestinal epithelial cells, IEC-18 and their K-ras transformed clones. Lovastatin induced morphologic changes (cell rounding and detachment) and apoptosis that were not influenced by K-ras mutations, but were prevented by geranylgeranyl-pyrophosphate or by mevalonate. Clostridium difficile toxin B, which directly inactivates rho, induced similar morphologic changes and apoptosis. Cycloheximide prevented these effects of lovastatin, but not C. difficile toxin B. Lovastatin decreased the amounts of membrane bound rhoA and rhoB. Cycloheximide and geranylgeranyl pyrophosphate prevented lovastatin induced morphologic changes and apoptosis but did not inhibit lovastatin-induced changes in membrane translocation of rho. Our data suggest that lovastatin induces morphologic changes and apoptosis by inhibiting geranylgeranylation of small GTPases of the rho family and thereby inactivating them. Restoration of membrane translocation of rho is not necessary for preventing lovastatin-induced morphologic changes or apoptosis. PMID- 11895869 TI - Enterococcus faecalis produces extracellular superoxide and hydrogen peroxide that damages colonic epithelial cell DNA. AB - Enterococcus faecalis is a commensal microorganism of the human intestinal tract that produces substantial extracellular superoxide (O(-)(2)), and derivative reactive oxygen species such as H(2)O(2) and hydroxyl radical, through autoxidation of membrane-associated demethylmenaquinone. Because these oxidants may be important as a cause of chromosomal instability (CIN) associated with sporadic adenomatous polyps and colorectal cancer, the ability of E.faecalis to damage eukaryotic cell DNA was examined using the alkaline lysis single cell gel electrophoresis (comet) assay. Both Chinese hamster ovary and HT-29 intestinal epithelial cells showed increased DNA damage after co-incubation with wild-type E. faecalis strain OG1RF, but not a transposon-inactivated mutant with attenuated extracellular O(-)(2) production. E. faecalis-mediated DNA damage was prevented by catalase, but not manganese superoxide dismutase, indicating H(2)O(2) arising from O(-)(2) was the genotoxin. In a rat model of intestinal colonization, OG1RF resulted in significantly higher stool concentrations of H(2)O(2) and 5,5 dimethyl-1-pyrroline N-oxide adducts of hydroxyl and thiyl radicals, as identified by electron spin resonance-spin trapping, compared with rats colonized with a mutant strain having attenuated O(-)(2) production. Using the comet assay, luminal cells from the colon of rats colonized with O(-)(2)-producing E. faecalis showed significantly increased DNA damage compared with control rats colonized with the mutant. These findings suggest a potentially profound role for extracellular free radical production by E. faecalis in promoting CIN associated with sporadic adenomatous polyps and colorectal cancer. PMID- 11895870 TI - The influence of folate and multivitamin use on the familial risk of colon cancer in women. AB - Low intake of folate and methionine and heavy alcohol consumption have been associated with an increased overall risk of colon cancer, possibly related to their role in methylation pathways. We estimated the relative risk (RR) of colon cancer according to a history of colorectal cancer in a first-degree relative and categories of folate, methionine, and alcohol intake in a prospective cohort study of 88,758 women who completed family history and detailed food frequency questionnaires. During 16 years of follow-up, colon cancer was diagnosed in 535 women. The inverse association of folic acid with colon cancer risk was greater in women with a family history. Compared with women who consumed 200 microg or less of folic acid/day, the age-adjusted RR of colon cancer for those who consumed >400 microg/day was 0.81 (95% confidence interval, 0.62-1.07) in women without a family history of colorectal cancer and 0.48 (95% confidence interval, 0.28-0.83) in women with a family history (P for interaction = 0.02). The influence of family history was markedly diminished by use of multivitamins containing folic acid (P for interaction = 0.04). High levels of dietary methionine also reduced the effect of family history (P for interaction = 0.05), whereas moderate to heavy alcohol consumption increased the risk associated with family history (P for interaction = 0.004). Other risk factors for colorectal cancer did not significantly modify the influence of family history. Our results suggest that higher intake of folate and methionine, regular use of multivitamins containing folate, and avoidance of moderate to heavy alcohol consumption may diminish the excess risk of colon cancer associated with a family history of the disease. PMID- 11895871 TI - The relation of reproductive factors to mortality from breast cancer. AB - Young women with breast cancer have been reported to have an increased risk of dying from their disease if they have given birth in <2 years before diagnosis. The prognostic factors associated with the tumors of these women have not been thoroughly studied. We examined the tumors of the women who had a recent birth and compared the tumor characteristics with those of women who were nulliparous or had given birth > or =5 years before diagnosis. A follow-up study was conducted of 1174 women <45 years old whose invasive ductal breast cancer was diagnosed from January 1983 to December 1992 in three counties of western Washington. These women had participated previously in a population-based, case control study. Mean follow-up time was 105.4 months. Histological slides were collected for 79.1% of the tumors and reviewed by the study pathologist. Using immunoperoxidase assays, tumor tissue was tested for prognostic markers for 70.4% of the tumors from the women. Cox proportional hazards models were used to estimate the relative risk of dying from breast cancer associated with reproductive events. Logistic regression was used to obtain estimates of the association between various reproductive factors and tumor characteristics. At the end of follow-up, 48.2% of the women (n = 83) whose last birth occurred in < 2 years of diagnosis had died, compared with 23.3% of nulliparous women (n = 189) and 24.4% of the women (n = 661) whose last birth was > or =5 years before diagnosis. The tumors of the women with a recent birth (<2 years before diagnosis) were more likely to be progesterone receptor negative, odds ratio (OR) = 2.2, 95% confidence interval (CI) = 1.2-3.9, to be p53 positive, OR = 2.6, 95% CI = 1.5-4.7, to be of high histological grade, OR = 5.9, 95% CI = 1.7-20.1, to have high mitotic count, OR = 2.2, 95% CI = 1.4-4.4, to be node positive, OR = 2.1, 95% CI = 1.3-3.5, to have a high S phase fraction, OR = 2.3, 95% CI = 1.1 4.8, and to have a high American Joint Committee on Cancer stage (III+), OR = 2.8, 95% CI 1.3-5.8, compared with the tumors of nulliparous women. After adjusting for tumor characteristics and treatment, the risk of mortality associated with a birth in < 2 years of diagnosis of breast cancer remained an independent predictor of mortality, hazard radio (HR) = 2.7, 95% CI = 1.6-4.3. Our study provides evidence that reproductive factors influence the biological behavior of breast cancer in young women and prognosis. Clinicians need to be aware that women who have delivered a child in < 2 years before diagnosis are at increased risk of having tumors with especially adverse prognostic profiles and have a poorer survival rate than women who are nulliparous or whose last birth was some years in the past. PMID- 11895872 TI - A polymorphism in the CYP17 gene and risk of prostate cancer. AB - Steroid hormones are important in the etiology and progression of prostate cancer, and expression of genes involved in hormone production may alter susceptibility. One such gene is CYP17, which encodes the cytochrome P450c17a enzyme responsible for the biosynthesis of testosterone. A T to C transition (A2 allele) in the 5' promoter region of the gene is hypothesized to increase the rate of gene transcription, increase androgen production, and thereby increase risk of prostate cancer. To test this hypothesis, germ-line DNA samples from a large population-based study of incident prostate cancer cases (n = 590) and controls (n = 538) of similar age without the disease were genotyped. The frequency of the A2 allele was similar in cases and controls. Compared with men with the A1/A1 genotype, the adjusted odds ratio was 0.81 for the A1/A2 and 0.87 for the A2/A2 genotype. Risk estimates did not vary substantially by age or race. However, stratification by family history of prostate cancer revealed that among white men with an affected first-degree relative, homozygotes for the A2 allele had a significant elevation in risk (odds ratio = 19.2; 95% confidence interval, 2.2-157.4) compared with men who were homozygous for the A1 allele (interaction P = 0.0005). These results suggest that the CYP17 A2/A2 genotype predicts susceptibility to prostate cancer in white men with a family history of the disease. It is also possible that CYP17 interacts with other genes that influence risk of familial prostate cancer. PMID- 11895873 TI - Recruiting adolescents into genetic studies of smoking behavior. AB - The goal of this study is to describe the process of establishing a longitudinal cohort to study genetic, psychological, and social predictors of adolescent smoking. Parents of eligible adolescents were approached for their consent via mail. Seventy-two percent of parents (n = 1533 of 2120) provided a response regarding their teens' participation. Among those who provided a response, 75% (1151) agreed to allow their teen to participate in the research yielding an overall parental consent rate of 54%. Compared with parents who consented to their teens' participation, parents who declined were less educated (89% had greater than a high school education compared with 69% of those who did not provide consent), less likely to be Caucasian (68 versus 48%), and less likely to report having ever even experimented with smoking (71 versus 60%). The most frequently reported reasons parents gave for declining consent included lack of interest and confidentiality concerns. A logistic regression model predicting consent to participate revealed a significant race by education interaction, indicating that among Caucasian parents, those with an education beyond high school were over two times more likely to provide consent compared with Caucasian parents with a high school education or less (odds ratio = 2.43; confidence interval = 1.37-4.32, P = 0.003). PMID- 11895874 TI - Overnight urinary isoflavone excretion in a population of women living in the United States, and its relationship to isoflavone intake. AB - Dietary isoflavones are biologically active in humans, but few observational data exist on the relationship between isoflavone intake and excretion in Western populations. We examined associations between self-reported soy intakes and overnight urinary isoflavone excretion in a population-based sample of western Washington State women, and we investigated the usefulness of one versus two overnight urine samples, collected 48 h apart, as a biomarker of intake. Isoflavones (genistein, daidzein, O-desmethylangolensin, and equol) were measured in two overnight urine collections from 363 women recruited from a health maintenance organization. Soy food intakes were assessed using two 1-day diet records completed on each day prior to the urine collections and a food frequency questionnaire (FFQ) that had been completed by 312 of the women with regard to their dietary habits 3.5 years (range, 2-5 years) before the urine collections. Twenty-one percent of the women consumed soy on either day of the diet recall, and 13% and 34% of the women consumed soy at least once a week or at least once a month, respectively, according to the FFQ. Women who consumed soy at either of the two diet recalls or at the FFQ (at least once a week or at least once a month) had a significantly higher urinary excretion of isoflavones than women who did not consume soy (P < 0.01). Among women who consumed soy at either of the two diet recalls or at the FFQ (soy consumed at least once a month), isoflavone intake and excretion correlated significantly (P < 0.01). Excretion of the individual isoflavones correlated significantly between the two urine samples collected 48 h apart (genistein, r = 0.41 and P < 0.001; daidzein, r = 0.30 and P < 0.001; O-desmethylangolensin, r = 0.46 and P < 0.001; equol, r = 0.60 and P < 0.001). Differences between soy consumers and nonconsumers and associations between intakes and excretion remained significant whether one or both urine collections were considered. Measuring isoflavone excretion in one overnight urine collection serves as a biomarker of recent or past isoflavone intake, even in populations whose intake of soy foods is relatively low. PMID- 11895875 TI - Dietary fat, fat subtypes, and breast cancer risk: lack of an association among postmenopausal women with no history of benign breast disease. AB - A recent study among 13,707 postmenopausal women without benign breast disease (BBD) from the Breast Cancer Detection Demonstration Project (BCDDP) cohort found breast cancer risk associated with greater total fat, unsaturated fat, and oleic acid intake. We assessed the associations between cumulative averaged dietary intake from 1980, 1984, 1986, and 1990 with breast cancer risk through 1994 among 44,697 postmenopausal participants without BBD in the Nurses' Health Study (NHS). Multivariate Cox proportional hazard models, with age as the time variable, provided the estimated rate ratios (RRs) with 95% confidence intervals (CIs) from the 14 years of follow-up and the 1,071 breast cancer cases. In the Nurses' Health Study, breast cancer rates over the time period from 1980 to 1994 did not increase significantly with greater total fat [quintile (Q) 5 versus Q1 RR, 0.94; 95% CI, 0.77-1.15], saturated fat (RR(Q5 to Q1), 0.88; 95% CI, 0.70-1.12), unsaturated fat (RR(Q5 to Q1), 1.16; 95% CI, 0.92-1.46), oleic acid (RR(Q5 to Q1), 1.13; 95% CI, 0.81-1.57), linoleic acid (RR(Q5 to Q1), 0.93; 95% CI, 0.74 1.16), trans fatty acid (RR(Q5 to Q1), 0.9184; 95% CI, 0.73-1.13), or energy intake (RR(Q5 to Q1), 0.81; 95% CI, 0.67-0.99). A parallel analysis restricted to the same time period as the BCDDP study did not differ substantially. We found no increase in the rate of breast cancer with greater intake of dietary fat and fat subtypes among postmenopausal women without a history of BBD. PMID- 11895876 TI - Detection of malondialdehyde DNA adducts in human colorectal mucosa: relationship with diet and the presence of adenomas. AB - Colorectal biopsies from normal mucosa of participants in the United Kingdom Flexible Sigmoidoscopy Trial and European Prospective Investigation on Cancer (EPIC; n = 162) were analyzed for the presence of malondialdehyde-deoxyguanosine (M(1)-dG), a DNA adduct derived from lipid peroxidation. The aim was to investigate whether dietary factors can modulate M(1)-dG levels and whether M(1) dG in normal mucosa is a risk factor for colorectal adenomas. Samples were analyzed using a sensitive immunoblot blot assay. This study has shown for the first time that M(1)-dG is present in human colorectal tissue. M(1)-dG levels ranged from undetectable (n = 13) to 12.23 per 10(7) total bases. Mean levels were 4.3 +/- 3 and 4.6 +/- 2.9 per 10(7) total bases in men and women, respectively. In men, there were positive associations of adduct levels with height and age, and inverse associations with body mass index. Legumes, fruit, salad, and whole meal bread were inversely associated with M(1)-dG adducts, whereas consumption of offal, white meat, beer, and alcohol were positively associated with elevated levels. In women, there was an inverse association of the adduct with the ratio of polyunsaturated:saturated fatty acids (P = 0.019) and a weak positive correlation with saturated fat (P < 0.061). When levels of adducts were compared in individuals with and without adenomas, there was a trend for higher levels in individuals presenting with adenomas especially in the highest category of M(1)-dG adducts (P < 0.005). PMID- 11895877 TI - A dose-finding study of aspirin for chemoprevention utilizing rectal mucosal prostaglandin E(2) levels as a biomarker. AB - Epidemiological and experimental evidence indicates that aspirin can protect against colorectal cancer. Aspirin inhibits cyclooxygenase enzymes and blocks prostaglandin (PG) biosynthesis. Using rectal PGE(2) levels as a mucosal biomarker, we sought to determine the optimal aspirin dose that would significantly suppress PGE(2) levels for chemoprevention trials. We conducted a randomized, double-blinded study in 60 subjects with prior sporadic colorectal adenoma(s) and evaluated three aspirin doses (81, 325, and 650 mg) or placebo taken daily for 4 weeks. PGE(2) levels in rectal biopsies performed at baseline and week 4 were analyzed by competitive immunoassay. Plasma salicylate levels, pill counts, and subject calendars were used to assess compliance. The 81-mg aspirin dose significantly suppressed PGE(2) levels relative to placebo (P = 0.005) and did so to an equivalent extent as did higher doses (P > 0.4) in evaluable subjects (n = 55) over a 4-week treatment period. Serum salicylate levels were associated with aspirin dose (P = 0.0002). Pill counts and calendars indicated that >98% of doses were taken by all subjects. No adverse events occurred in this short-term study. The 81-mg daily aspirin dose suppressed PGE(2) levels to an equivalent extent as did the 650-mg dose and supports the use of this dose for chemoprevention trials. PMID- 11895878 TI - Post-study aspirin intake and factors motivating participation in a colorectal cancer chemoprevention trial. AB - We conducted an exploratory, cross-sectional study examining motivators for study participation and post-study aspirin intake in a chemoprevention trial. The parent clinical trial aimed to determine the optimal aspirin dose for colorectal cancer chemoprevention using prostaglandin E(2) as a mucosal biomarker. This trial was randomized and double-blinded in 60 subjects with prior sporadic colorectal adenoma(s) and evaluated three aspirin doses or placebo taken once daily for 4 weeks. A cross-section of 55 evaluable participants who completed the chemoprevention trial were mailed a 16-item, self-administered questionnaire evaluating subject demographics, motivational factors, and health-related behaviors within the framework of Pender's Health Promotion Model (HPM). Forty three (78%) of 55 participants returned the questionnaire. The most important motivators for study participation were altruistic, i.e., a desire to help future generations at risk of colorectal cancer and personal factors including a desire to reduce one's own risk. Nineteen (44%) of 43 respondents reported that they chose to take daily aspirin post-study without knowledge of study results. At a mean follow-up of 17.3 months, 18 of these 19 subjects continued to take aspirin regularly. Regular use of vitamin supplements pre-study was found to correlate with post-study aspirin use (Mann-Whitney U test, U = 154.0; P = 0.04). We demonstrate, for the first time, that participation in a chemoprevention study can influence the decision to continue the study drug, if available, to reduce perceived cancer risk. Continued post-study aspirin intake indicates an impact of study participation on a health-related behavior and underscores the importance of patient education to guide such decision-making. PMID- 11895879 TI - Correlations of blood lead with DNA-protein cross-links and sister chromatid exchanges in lead workers. AB - Levels of sister chromatid exchanges (SCEs), high-SCE frequency cells (HFCs), DNA protein cross-links (DPCs), blood lead (BLL), and zinc protoporphyrin (ZPP) were measured in peripheral blood from three groups. The lead workers were divided into two groups: a high BLL group (> or =15 microg/dl) and a low BLL group (<15 microg/dl). The control subjects were selected from an area that had not been contaminated with lead and had normal BLL and ZPP levels. In addition, exposure to airborne lead was measured for 11 lead workers, and the time-weighted average was shown to range from 0.19 to 10.32 mg/m(3). The BLL levels of 9 of 11 workers were >15 microg/dl, of which, 3 exceeded current exposure limits (> or =40 microg/dl). The BLL levels of all 11 controls were < 15 microg/dl. The average SCE and DPC values for the workers were 6.1 SCEs/cell and 1.9%, which were significantly higher (P < 0.01, Wilcoxon's test) than the value of 5.2 SCEs/cell and 1.1% for the control subjects. Lead workers had significantly higher BLL and ZPP levels than did the controls. Statistically significant increases in DPCs, SCEs, and HFCs were observed for the high-BLL group compared with the control group. The results of this study suggest that DPCs, SCEs, and HFCs are reliable biomarkers for monitoring workers exposed to lead and clearly indicate health effects from occupational exposure to lead. PMID- 11895880 TI - Hierarchical clustering of lung cancer cell lines using DNA methylation markers. AB - Recent analyses of global and gene-specific methylation patterns in cancer cells have suggested that cancers from different organs demonstrate distinct patterns of CpG island hypermethylation. Although certain CpG islands are frequently methylated in many different kinds of cancer, others are methylated only in specific tumor types. Because distinct patterns of CpG island hypermethylation can be seen in tumors from different organs, it seems likely that histological subtypes of cancer within a given organ may exhibit distinct methylation patterns as well. The goal of our study was to determine whether the patterns of CpG island hypermethylation could be used to distinguish between different histological subtypes of lung cancer. We analyzed the methylation status of 23 loci in 91 lung cancer cell lines using the quantitative real-time PCR method MethyLight. Genes PTGS2 (COX2), CALCA, MTHFR, ESR1, MGMT, MYOD1, and APC showed statistically significant differences in the level of CpG island methylation between small cell lung cancer (SCLC) and non-small cell lung cancer cell lines (NSCLC). Hierarchical clustering using a panel consisting of these seven loci yielded two major groups, one of which contained 78% of the SCLC lines. Within this group, a large cluster consisted almost exclusively of SCLC cell lines. Our results show that DNA methylation patterns differ between NSCLC and SCLC cell lines and suggest that these patterns could be developed into a powerful molecular marker to achieve accurate diagnosis of lung cancer. PMID- 11895881 TI - Adrenalectomy abrogates reduction of 12-O-tetradecanoylphorbol-13-acetate-induced extracellular signal-regulated protein kinase activity in the epidermis of dietary energy-restricted SENCAR mice: implications of glucocorticoid hormone. AB - Previous studies in this laboratory demonstrated that dietary energy restriction (DER), a potent inhibitor of skin carcinogenesis, markedly suppressed 12-O tetradecanoylphorbol-13-acetate (TPA)-induced extracellular signal-regulated protein kinase (ERK) activity in mouse epidermis. Intact adrenal glands have been reported to be important in the inhibition of skin tumor promotion by food restriction. We investigated the role of adrenal glands and corticosterone in the DER effect on ERK activity. Female SENCAR mice, either sham operated or adrenalectomized (Adx), were prefed with 40% DER diet for 8-10 weeks and treated with a single TPA treatment or twice weekly TPA after initiation by 7,12 dimethylbenz(a)anthracene (DMBA). ERK activity measured 1 h after the last TPA treatment was significantly inhibited by DER in sham-operated mice, whereas the ERK inhibitory effect of DER was completely abolished in Adx mice. In a parallel study, Adx mice were provided with 60 microg/ml corticosterone in the drinking water to test the hypothesis that corticosterone from the adrenal gland plays a key role in the DER inhibition of ERK in mice with intact adrenal glands. There was an overall elevated plasma corticosterone level in DER mice compared with AL mice. Adx decreased the plasma corticosterone level and abrogated the DER inhibition of ERK activity. Addition of corticosterone in the drinking water restored plasma hormone level and markedly decreased TPA-induced ERK activity in Adx mice (P < 0.05). These results provide strong evidence that intact adrenal glands are essential for the DER inhibition of ERK induction by TPA and that corticosterone plays a critical role in the DER blockage of ERK induction. PMID- 11895882 TI - The expression of a variant prostate-specific antigen in human prostate. AB - Although a splicing variant of prostate specific antigen (PSA-v) mRNA has been described previously, whether its protein (PSA-v or PSA-related protein 1, i.e., PSA-RP1) is actually expressed in human prostate cells remains elusive. We report that PSA-v protein is expressed in prostatic epithelia of both cancerous and benign tissues. Also, secreted PSA-v can be detected in the medium of a prostate cancer (PCa) cell line. Consistently, PSA-v mRNA is exclusively expressed in benign luminal epithelia and cancer cells of the prostate by in situ hybridization. Northern analysis of a cohort of 51 pairs of RNA samples from microdissected tissues showed that PSA-v mRNA levels remained constant in both benign and cancerous tissues, whereas PSA levels declined in cancerous areas. Our result suggests that it would be feasible to develop proper immunoassays for PSA v to test whether PSA-v could have some clinical utility. PMID- 11895883 TI - Relationship of exposure to coke-oven emissions and urinary metabolites of benzo(a)pyrene and pyrene in coke-oven workers. AB - Coke-oven workers are occupationally exposed to a high concentration of polycyclic aromatic hydrocarbons (PAH). r-7,t-8,9,c-10-Tetrahydroxy-7,8,9,10 tetrahydrobenzo(a)pyrene (trans-anti-BaP-tetraol) and 1-hydroxypyrene (1-OHP) are urinary metabolites of benzo(a)pyrene and pyrene, respectively. In this study, we investigated the relationship among individual air exposure to benzene soluble fraction (BSF) of total particulates, as a surrogate marker of ambient PAH exposures, and urinary trans-anti-BaP-tetraol and 1-OHP concentrations in coke oven workers at a steel plant in Taiwan. Fifty-seven subjects, including 41 male workers who work in one coke-oven plant and 16 men (referents) from an administrative area, were studied. The mean trans-anti-BaP-tetraol and 1-OHP concentrations (mean +/- SD) were 0.4 +/- 0.3 nmol/mol creatinine and 9.7 +/- 21.6 micromol/mol creatinine, respectively, in coke-oven workers. These levels were significantly higher than those in referents (0.03 +/- 0.03 nmol/mole creatinine, P < 0.001 and 0.4 +/- 0.2 micromol/mol creatinine, P < 0.01, respectively). Urinary trans-anti-BaP-tetraol concentrations were significantly and positively correlated with individual average BSF and urinary 1-OHP concentrations. That is, the higher the urinary trans-anti-BaP-tetraol concentrations, the more ambient BSF exposure and urinary 1-OHP concentrations (Spearman correlation coefficients r = 0.68 and 0.70, respectively; P < 0.0001; n = 57). These findings suggest that urinary 1-OHP and trans-anti-BaP-tetraol might be considered as potential biomarkers for the assessment of uptake of known PAH carcinogens in the air. PMID- 11895884 TI - Analysis of DNA and hemoglobin adducts and sister chromatid exchanges in a human population occupationally exposed to propylene oxide: a pilot study. AB - Propylene oxide (PO), a simple alkylating agent used in the chemical industry, is weakly genotoxic and induces nasal cavity tumors in rodents on inhalation at high air concentrations. DNA adducts, hemoglobin adducts, and sister chromatid exchanges (SCE) were analyzed as biomarkers of exposure in a group of eight PO exposed workers and eight nonexposed subjects. 1-2-Hydroxypropyladenine (1-HP adenine) in DNA of WBCs was analyzed using a hypersensitive (32)P-postlabeling assay. HP-valine in hemoglobin was measured using gas chromatography/tandem mass spectrometry. Air measurements indicated PO levels in the range of 1-7 ppm. All three biomarkers showed significantly increased levels in the exposed workers. 1 HP-adenine was recorded in seven of the exposed workers (mean 0.66 mol/10(9) mol nucleotides) but was not detected in any of the control subjects. HP-valine was found in all subjects (means of 2.7 and 0.006 pmol/mg globin in exposed workers and controls, respectively). The average frequencies of SCE were 3.7/cell in exposed workers and 2.0/cell in controls, respectively. DNA and hemoglobin adducts were correlated (r = 0.887), as well as DNA adducts and SCE (r = 0.792) and hemoglobin adducts and SCE (r = 0.762). The present study is the first demonstrating PO-DNA adducts in human individuals. It is also the first study indicating cytogenetic effects in humans from PO exposure, although confounding effects from other sources cannot be excluded. PMID- 11895885 TI - Serum insulin-like growth factor I (IGF-I) concentration in men is not associated with the cytosine-adenosine repeat polymorphism of the IGF-I gene. PMID- 11895886 TI - No association between smoking and the presence of tobacco-specific nitrosamine metabolites in ovarian follicular fluid. PMID- 11895887 TI - Correspondence re: Chokkalingam A. et al., Insulin-like growth factors and prostate cancer: a population-based case-control study in China. Cancer Epidemiol. Biomark. Prev., 10: 421-427, 2001. PMID- 11895888 TI - Correspondence re: Weiderpass et al., Alcoholism and risk of cancer of cervix uteri, vagina, and vulva. Cancer Epidemiol. Biomark. Prev., 10: 899-901, 2001. PMID- 11895889 TI - Modulating multidrug resistance: can we target this therapy? PMID- 11895890 TI - A view to a kill: signals triggering cytotoxicity. AB - Tumor immunity requires the participation of lymphocyte effector cells that display powerful processes to destroy malignant cells. Natural killer (NK) cells and CTLs, once activated, use the same lytic processes for mediating target cell death. However, they are triggered through distinctly separate antigen receptors. NK cells are currently known to express at least three families of receptors unrelated to the T cell receptor, i.e., NKG2, KIR, and NCR, to mediate cytotoxicity. This review provides a view to a kill, bringing together a unifying concept for a common signal pathway that dictates lytic function. What emerges is a specific Syk/Zap70 --> PI3K --> Rac --> PAK --> MEK --> ERK signal cascade triggered by target cell recognition, which is responsible for mobilizing the lytic granules containing perforin and granzyme B toward the contacted target cell. PMID- 11895891 TI - Current perspectives on the clinical experience, pharmacology, and continued development of the camptothecins. AB - The camptothecins are a maturing class of anticancer agents. In this article, we review the pharmacology and antitumor activity of the camptothecin analogues that are approved for clinical use and those investigational agents undergoing clinical evaluation. Camptothecin is a naturally occurring cytotoxic alkaloid that has a unique intracellular target, topoisomerase I, a nuclear enzyme that reduces the torsional stress of supercoiled DNA during the replication, recombination, transcription, and repair of DNA. Topotecan and irinotecan are synthetic analogues designed to facilitate parenteral administration of the active lactone form of the compound by introducing functional groups to enhance solubility. They are now well-established components in the chemotherapeutic management of several neoplasms. Topotecan has modest activity in patients treated previously with ovarian and small cell lung cancer and is currently approved for use in the United States as second-line therapy in these diseases. Preliminary evidence of activity against hematological malignancies is also promising. Irinotecan is a prodrug that undergoes enzymatic conversion to the biologically active metabolite 7-ethyl-10-hydroxy-camptothecin. It is presently the treatment of choice when used in combination with fluoropyrimidines as first line therapy for patients with advanced colorectal cancer or as a single agent after failure of 5-fluorouracil-based chemotherapy. Encouraging preliminary results suggest that irinotecan may have an increasing role in the treatment of other solid tumors, including small and non-small cell lung cancer, cervical cancer, ovarian cancer, gastric cancer, and malignant gliomas. Several additional camptothecin analogues are in various stages of clinical development, including 9 aminocamptothecin, 9-nitrocamptothecin, 7-(4-methylpiperazinomethylene)-10,11 ethylenedioxy-20(S)-camptothecin, exatecan mesylate, and karenitecin. Efforts to further optimize therapeutic effectiveness through drug delivery strategies that prolong tumor exposure to these S phase-specific agents, such as improving oral bioavailability through structure modification and innovative formulation approaches, alternative parenteral dosage forms, and administration schedules, are being actively pursued. Combining camptothecins with other anticancer drugs and treatment modalities, as well as gaining a better understanding of the factors contributing to tumor sensitivity and resistance, continues to be the object of considerable interest. PMID- 11895892 TI - Histone deacetylase inhibitors: a new class of potential therapeutic agents for cancer treatment. PMID- 11895893 TI - Approval summary: letrozole in the treatment of postmenopausal women with advanced breast cancer. AB - Letrozole (Femara; Novartis Pharmaceuticals Corp., East Hanover, NJ) is a nonsteroidal inhibitor of aromatase enzyme complex. It inhibits the peripheral conversion of circulating androgens to estrogens. In postmenopausal women, letrozole decreases plasma concentrations of estradiol, estrone, and estrone sulfate by 75-95% from baseline with maximal suppression achieved within 2-3 days of treatment initiation. Suppression is dose related, with doses of >or=0.5 mg giving estrone and estrone sulfate values that were often below assay detection limits. At clinically used dosage, letrozole does not impair adrenal synthesis of glucocorticoids or aldosterone. In 1998, letrozole was approved by the United States Food and Drug Administration (FDA) for the treatment of advanced breast cancer in postmenopausal women, with hormone receptor positive or unknown breast cancer, who had failed one prior antiestrogen treatment (i.e., for "second-line" treatment). Approval was based on two randomized trials comparing tumor RRs of patients receiving 0.5 mg of letrozole, 2.5 mg of letrozole, and either megestrol acetate (MA) or aminoglutethimide. In the megestrol trial, 2.5 mg/day letrozole was superior to 0.5 mg of letrozole and MA (RRs 24, 13, and 16%, respectively), whereas in the aminoglutethimide trial, there was no significant difference in 2.5 mg of letrozole and 0.5 mg of letrozole RRs (20 and 17%). There was a trend toward RR superiority of 2.5 mg of letrozole over aminoglutethimide (P = 0.06). Letrozole (2.5 mg) was the dose chosen for comparison with tamoxifen in the first line setting. In July 2000, a marketing application for first-line letrozole treatment of postmenopausal women with hormone receptor positive or hormone receptor unknown locally advanced or metastatic breast cancer was submitted to the FDA. A single double-blind, double dummy, randomized, and multicenter trial compared 2.5 mg of letrozole to 20 mg of tamoxifen (456 patients/arm). Letrozole was superior to tamoxifen with regard to time to progression (TTP) and objective response rate (RR). The median TTP for letrozole treatment was 9.9 months [95% confidence interval (CI) 9.1-12.2] versus 6.2 months (95% CI 5.8-8.5) for tamoxifen, P = 0.0001, hazard ratio 0.713, (95% CI 0.61-0.84). RR was 32% for letrozole versus 21% for tamoxifen (odds ratio 1.74, 95% CI 1.29-2.34, P = 0.0003). Preliminary survival data (survival data are still blinded) indicate that letrozole is unlikely to be worse than tamoxifen. Both treatments were similarly tolerated. On the basis of these results, the United States FDA approved letrozole tablets, 2.5 mg/day, for first-line treatment of postmenopausal women with hormone receptor-positive or hormone receptor-unknown locally advanced or metastatic breast cancer. The manufacturer made a commitment to provide updated information on survival. PMID- 11895894 TI - Safety and efficacy of the multidrug resistance inhibitor Incel (biricodar; VX 710) in combination with paclitaxel for advanced breast cancer refractory to paclitaxel. AB - PURPOSE: VX-710 (biricodar, Incel) restores drug sensitivity to P-glycoprotein (MDR1) and multidrug resistance-associated protein (MRP1)-expressing cells. This Phase II study evaluated the safety/tolerability, pharmacokinetics, and efficacy of VX-710 plus paclitaxel in women with locally advanced or metastatic breast cancer who were refractory to prior paclitaxel therapy. EXPERIMENTAL DESIGN: Eligible patients had paclitaxel-refractory disease defined as progressive disease after a minimum of two cycles of paclitaxel (weekly or 3-week schedule) or relapsed/progressive disease within 6 months of prior paclitaxel therapy. Patients received 80 mg/m(2) paclitaxel over 3 h starting 4 h after initiation of a 24-h continuous i.v. infusion of 120 mg/m(2)/h VX-710. Cycles were repeated every 3 weeks. RESULTS: Thirty-seven patients received study treatment and 35 were evaluable for response. VX-710 + paclitaxel therapy was generally well tolerated. Myelosuppression was the principal toxicity, with a median nadir ANC cycle 1 of 0.76 x 10(9) cells/liter and a 40% overall incidence of Grade 4 neutropenia. Nonhematological side effects (asthenia, paresthesia, headache, myalgia, nausea, and diarrhea) were generally mild to moderate and reversible. Paclitaxel AUC (16.8 +/- 5.0 microg x h/ml) and clearance (5.1 +/- 1.3 liters/h/m(2)) during the first treatment cycle were comparable with standard 175 mg/m(2) paclitaxel administered in a 3-h schedule. Four patients achieved partial responses (three of the four had progressive disease on prior paclitaxel) with a mean response duration of 5.5 months. CONCLUSIONS: The 11.4% (4 of 35) objective response rate observed in this study suggests that VX-710 can resensitize a subgroup of paclitaxel-refractory patients to paclitaxel. The safety and pharmacokinetics of the VX-710/pacitaxel regimen support further evaluation in breast cancer patients with initial paclitaxel therapy to prevent emergence of the MDR phenotype in recurrent disease. PMID- 11895895 TI - Phase I trial of BCL-2 antisense oligonucleotide (G3139) administered by continuous intravenous infusion in patients with advanced cancer. AB - PURPOSE: To evaluate the safety and pharmacokinetics of BCL-2 antisense oligonucleotide (G3139) administered by prolonged i.v. infusion in patients with advanced cancer. EXPERIMENTAL DESIGN: A total of 35 patients was treated in cohorts of 3-6 with 0.6-6.9 mg/kg/day of BCL-2 antisense oligonucleotide as a continuous infusion for 14 or 21 days. Plasma levels of intact antisense oligonucleotide were measured in all patients. RESULTS: G3139 was generally well tolerated. At the highest dose level examined in this study (6.9 mg/kg/day), fatigue and transient reversible elevations of serum transaminases (grades 2-3) became apparent after >or=7 days of treatment. Both reactions were believed to be drug related. Pharmacokinetic analyses showed that steady-state plasma concentrations of G3139 were reached approximately 10 h after starting the infusion and increased linearly across the range of doses administered or=2 prior regimens). RESULTS: The maximum tolerated dose of the combination was 25 mg/m(2) doxorubicin and 5.25 mg/m(2) topotecan (1.75 mg/m(2)/day x 3). Neutropenia was the dose-limiting toxicity. Attempts to further escalate the dose using 5 microg/kg granulocyte colony-stimulating factor proved unsuccessful because of thrombocytopenia. Among the 17 patients who were evaluable for response, 6 had a partial response, and 4 showed evidence of disease stabilization. The partial responses occurred in patients with small cell lung cancer (3 of 7), non-small cell lung cancer (1 of 6), esophageal adenocarcinoma (1 of 2), and ovarian carcinoma (1 of 1), and it lasted for 3-6 months. Administration of doxorubicin 2 days before topotecan did not alter topotecan pharmacokinetics. Changes in topoisomerase mRNA levels were observed during chemotherapy. CONCLUSIONS: The sequential combination of doxorubicin followed by topotecan is highly active in several chemotherapy refractory long, ovary, and esophageal cancers. Despite significant neutropenia, toxicity is manageable and well tolerated. Phase II trials to further evaluate the efficacy of this promising combination regimen against non-Hodgkin's lymphoma and lung cancer have been initiated. PMID- 11895898 TI - Association of high-dose cyclophosphamide, cisplatin, and carmustine pharmacokinetics with survival, toxicity, and dosing weight in patients with primary breast cancer. AB - This report investigates relationships between the pharmacokinetics and pharmacodynamics of high-dose alkylators used for the treatment of primary breast cancer. Eighty-five women with primary breast cancer involving >or=10 lymph nodes received four cycles of standard-dose chemotherapy followed by a high-dose regimen consisting of: cyclophosphamide (1875 mg/m(2) once daily x 3), cisplatin (165 mg/m(2) given over 72 h), carmustine (600 mg/m(2)), and stem cell transplantation. Dosages were attenuated in patients whose body weight exceeded their calculated ideal weight by >20%. Pharmacokinetics of the high-dose chemotherapeutic agents were evaluated in each patient by collection and analysis of serial blood samples. Area under the concentration time curve (AUC) for cyclophosphamide and carmustine was highly variable (>10-fold inter-patient range) with coefficients of variation > 50%, in contrast to cisplatin exposures (2-fold range; coefficient of variation 12%). The dosing method for overweight patients resulted in significantly lower systemic exposure to cisplatin (P = 0.035). The parent cyclophosphamide clearance on the 1st day of administration was significantly higher in patients who experienced acute cardiac toxicity (n = 5; P = 0.011), whereas carmustine disposition was not found to be different in those developing pulmonary toxicity (n = 25; P = 0.96). Kaplan-Meier analysis (median follow-up of 5.9 years) demonstrated that patients with lower cyclophosphamide AUC (faster parent drug clearance to potentially cytotoxic compounds) survived longer (P = 0.031). Inter-individual differences in the pharmacokinetic disposition of high-dose chemotherapy may explain variability in both response and toxicity. Prospective strategies, which attempt to individualize AUC, should be evaluated in this setting. PMID- 11895899 TI - Phase I clinical and pharmacokinetic study of oral penclomedine (NSC 338720) in adults with advanced solid malignancy. AB - Penclomedine is a synthetic alpha-picoline derivative that has shown antitumor activity both in preclinical development and in Phase I work using an i.v. preparation. The main toxicities seen in those studies were dose dependent and mainly neurocerebellar, with hematological toxicity being far less severe. This Phase I trial of p.o. penclomedine was conducted to potentially alter the toxicity profile and to avoid the neurological side effects seen with i.v. penclomedine. Eligibility criteria included microscopic confirmation of a solid malignancy or lymphoma with a lack of effective anticancer therapy. Twenty patients were enrolled. The median age was 60.5 years, and the median performance status was one. All but one patient had received prior systemic therapy. The starting dose of penclomedine was 200 mg/m(2) p.o. for 5 days, and was escalated according to a traditional Fibonacci sequence until the maximum tolerated dose (MTD) was observed. No treatment-related deaths were observed during the study. The MTD was determined to be 800 mg/m(2) p.o. for 5 days. Dose-limiting toxicities included mainly neurocerebellar symptoms such as ataxia and dysmetria, but neurocortical symptoms, such as confusion, were seen as well. Myelosuppression was less common and resulted in the discontinuation of therapy in only two patients. Pharmacokinetics show that the observed MTD is consistent with the i.v. preparations, and that the bioavailability of p.o. penclomedine is 49 +/- 18%. This regimen can be considered for additional studies in patients with intracranial neoplasms, because good central nervous system penetration is evident. Further development of penclomedine metabolites, such as 4-O demethylpenclomedine, should be considered to minimize dose-limiting neurotoxicity. PMID- 11895900 TI - A Phase I study of bizelesin (NSC 615291) in patients with advanced solid tumors. AB - PURPOSE: To evaluate the toxicities, characterize the pharmacokinetics, and determine the maximum-tolerated dose of bizelesin administered once every 4 weeks. PATIENTS AND METHODS: Patients with advanced solid tumors received escalating doses of bizelesin as an i.v. push every 4 weeks. Pharmacokinetic studies were performed with the first treatment cycle. RESULTS: Nineteen eligible patients received a total of 54 courses of bizelesin at doses ranging from 0.1 to 1 microg/m(2). Dose-limiting toxicity of neutropenia was seen in 2 of 4 patients treated at the 1 microg/m(2) dose level. Nonhematological toxicity was generally mild with maximum toxicity being or=50% cyclin D3-positive lymphoma cells) cyclin D3 expression had a more advanced clinical stage (P = 0.003) and more often had extranodal disease in more than one site (P = 0.007) than patients with low cyclin D3 expression. Patients with high cyclin D3 expression had a significantly lower complete response rate (17% versus 74%; P < 0.001) and a shorter overall survival (3-year survival rate, 18% versus 74%; P < 0.001) than those with low cyclin D3 expression. Multivariate analyses that included cyclin D3 and the International Prognostic Index demonstrated that cyclin D3 expression had independent effects on the complete response rates and overall survival of the patients. In conclusion, high cyclin D3 expression is an independent predictive and prognostic factor associated with poor clinical outcome in patients with diffuse large B-cell lymphoma. PMID- 11895903 TI - Differential expression of growth factors in squamous cell carcinoma and precancerous lesions of the lung. AB - PURPOSE: This study was conducted to evaluate the clinical significance of the localization of epidermal growth factor receptor (EGF-r), transforming growth factor (TGF)-alpha, and erbB-2 in the development, progression and prognosis of squamous cell cancers (SCCs) of the lung. EXPERIMENTAL DESIGN: The localization of EGF-r, TGF-alpha, and erbB-2 was evaluated immunohistochemically in 60 archival specimens of SCC of the lung and in 60 lung specimens without cancer. To clarify the patterns of expression of EGF-r in these tumors, the patterns of expression of EGF-r in cells in culture were monitored after challenging normal human bronchial epithelial and SCC cell lines with either EGF or TGF-alpha at physiological concentrations. RESULTS AND CONCLUSIONS: The expression of EGF-r, erbB-2, and TGF-alpha were significantly higher in SCC and associated precancerous lesions than in the normal bronchial epithelium and hyperplastic lesions of noncancer specimens. A statistically significant stepwise increase in expression from uninvolved bronchial epithelium to precancerous lesions to SCC was observed with EGF-r and TGF-alpha. The localization of EGF-r in the cytoplasm (P = 0.04), but not in the membrane (P = 0.20), of SCCs was significantly associated with poor overall survival of subjects. To demonstrate the biological relevance of cytoplasmic expression of EGF-r, we noted that there was a prompt reduction in the membrane expression and a concomitant increase in cytoplasmic expression of EGF-r after adding either EGF or TGF-alpha to the cell culture medium. Overall, the study identified an involvement of EGF-r and TGF-alpha in the development of SCCs. The prognostic importance of EGF-r expression in the cytoplasm of lung cancer probably is an indication of the prognostic importance of trafficking of the EGF-r receptor between the Golgi apparatus and cell membranes and of internalization of EGF-r after an interaction with one of the EGF-r ligands at the cellular membrane surface. PMID- 11895904 TI - Clinical significance of poor CD3 response in head and neck cancer. AB - PURPOSE: The objective of our investigation was to prospectively study what the implications of an unresponsive CD3 receptor are on clinical outcome in advanced stage head and neck cancer patients. EXPERIMENTAL DESIGN: Lymph node mononuclear cells were purified from cancer patients and stimulated with immobilized anti-CD3 in vitro for 8 days. Two populations were identified, nonresponders (NRs) with [(3)H]thymidine-counts per min (cpm) <3500 and responders (Rs) with cpm >or=3500. NRs and Rs were prospectively followed for a minimum of 24 months, and clinical outcomes were compared. Postoperative complications, length of hospitalization, toxicities associated with chemotherapy or radiation therapy, survival, and disease-free status were measured. RESULTS: Twenty-six patients were followed, of which 19 Rs [[(3)H](X) = 37,819 +/- 24,979 cpm (mean proliferative count +/- SD)] and 7 NRs ([(3)H](X) = 1,375 +/- 1,102 cpm) were identified. There were no phenotypic differences in lymph node T-cell subpopulations (CD3, CD4, CD8, CD28, CD45RO) between groups. There was a 71% (5/7) incidence of recurrent cancer in NRs compared with 16% (3/19) in Rs; the median disease-free interval was significantly less in NRs (P = 0.03). The risk ratio of Rs to develop a recurrent cancer was 0.237 (95% confidence interval, 0.057-0.994), much less than for NRs. CONCLUSIONS: Patients with an unresponsive CD3 receptor as measured by in vitro response to anti-CD3 monoclonal antibodies had a significantly higher incidence of recurrent cancer. Analyses using Cox proportion hazards models demonstrated that CD3 response was the single greatest predictor of reduced disease-free interval. This is the first prospective study to confirm the importance of regional lymph node mononuclear cell CD3 receptor function in head and neck squamous cell carcinoma patients for tumor control. PMID- 11895906 TI - Immunohistochemical assessment of localization and frequency of micrometastases in lymph nodes of colorectal cancer. AB - PURPOSE: Micrometastases are often found in regional lymph nodes of colorectal cancer (CRC). The aim of this study is to examine the extent and distribution of such lymph nodes. EXPERIMENTAL DESIGN: We immunohistochemically assessed localization and frequency of micrometastases in 878 lymph nodes from 98 patients with CRC. The anatomical position of lymph nodes was defined as level 1 to level 3 according to distance from the main tumor. RESULTS: The frequency of micrometastasis increased through observation of the 4-microm-thick lymph node sections, from one to two to five slices. With five slices, micrometastasis was frequently and extensively present in 49.1, 35.7, and 53.3% patients of histologically node-negative patients, node-positive patients at level 1, and node-positive patients at level 2, respectively. We then assessed the value of the presence of micrometastasis in node-negative patients with regard to prognosis, but no significant impact was obtained. To examine the reproducibility of the results obtained with immunohistochemistry, serial sectioning (four consecutive slices at seven different levels) of lymph nodes was additionally performed in lymph nodes initially diagnosed as micrometastasis positive. Immunohistochemical detection revealed that the sectioning level highly affected the results. CONCLUSIONS: Our results indicated frequent presence of micrometastasis in lymph nodes of CRC and that micrometastasis in node-negative CRC patients did not help in predicting the outcome, in part because of the limited reproducibility with immunohistochemistry. PMID- 11895907 TI - Profound dihydropyrimidine dehydrogenase deficiency resulting from a novel compound heterozygote genotype. AB - A familial approach was used to elucidate the genetic determinants of profound and partial dihydropyrimidine dehydrogenase (DPD; EC 1.3.1.2) deficiency in an Alabama family. In 1988, our laboratory diagnosed profound DPD deficiency in a breast cancer patient with grade IV toxicity after cyclophosphamide/methotrexate/5-fluorouracil chemotherapy (R. B. Diasio et al., J. Clin. Investig., 81: 47-51, 1988). We now report the genetic analysis of archived genomic DNA that reveals that the proband was a compound heterozygote for two different mutations, one in each allele: (a) a G to A mutation in the GT 5' splicing recognition sequence of intron 14, which results in a 165-bp deletion (corresponding to exon 14) in the DPD mRNA (DPYD*2A); and (b) a T1679G mutation (now designated DPYD*13), which results in a I560S substitution. Sequence analysis revealed segregation of both mutations with the son and the daughter each inheriting one mutation. Phenotype analysis (DPD enzyme activity) confirmed that both children were partially DPD deficient. Plasma uracil and DPD mRNA levels were found to be within normal limits in both children. We conclude that profound DPD deficiency in the proband resulted from a combination of two mutations (one mutation in each allele) and that heterozygosity for either mutation results in partial DPD deficiency. Lastly, we identified two variant alleles reported previously as being associated with DPD enzyme deficiency [T85C resulting in a C29R substitution (DPYD*9A) and A496G (M166V) in a family member with normal DPD enzyme activity]. These data suggest that both variant alleles are unrelated to DPD deficiency and emphasize the need to perform detailed familial genotypic and phenotypic analysis while characterizing this pharmacogenetic syndrome. PMID- 11895908 TI - The association of p21((WAF-1/CIP1)) with progression to androgen-independent prostate cancer. AB - The molecular events leading to progression toward androgen-independent prostate cancer (AIPC) are not fully understood. The p21((WAF-1/CIP1)) (p21) gene has been identified as a key factor for the regulation of cell growth. The expression of p21 was examined by immunohistochemical studies in 105 prostate cancer samples: (a) 7 of 30 (23%) androgen-dependent tumors; and (b) 36 of 75 (48%) androgen independent tumors stained positive for p21 (P < 0.02). No association was found between p21 expression and p53, bcl-2, and the androgen receptor protein expression in bone metastases of patients with AIPC, whereas there was a significant association with a high Ki-67 index (P < 0.05). In 4 of 43 (9%) cases, tumors displayed a p53-negative, bcl-2-negative, and p21-positive phenotype. A xenograft mouse model of prostate cancer using the androgen responsive MDA PCa 2b prostate cancer cell line was used to study p21 expression after androgen deprivation and at relapse. Androgen deprivation reduced p21 expression to undetectable levels after 14 days. Tumor relapse, defining AIPC, was associated with increased expression of p21 to levels comparable with those found before castration. In this model, p21 expression at relapse was also correlated with a high Ki-67 index. In conclusion, p21 expression is associated with the progression to AIPC. A possible explanation involves a paracrine effect of p21 mediated by the release of mitogenic and antiapoptotic factors. Another explanation involves the regulation of p21 expression by the androgen receptor, which also suggests that p21 may have antiapoptotic function in prostate cancer. PMID- 11895905 TI - Autoimmunity to the M(r) 32,000 subunit of replication protein A in breast cancer. AB - PURPOSE: We sought to identify autoantigens recognized by antibodies in breast cancer patient sera with potential diagnostic or prognostic significance. EXPERIMENTAL DESIGN: Serum from a female breast cancer patient exhibiting a high titer antinuclear antibody was used to screen a HeLa cDNA expression library, leading to the cloning of a cDNA for the M(r) 32,000 subunit of replication protein A (RPA32). RPA32 expression and localization were assayed in autologous tumor by monoclonal antibody staining. A specific ELISA using recombinant protein was used to screen sera from 801 breast cancer patients and 65 controls. RESULTS: A relationship between anti-replication protein A (RPA) antibodies and the ductal breast carcinoma of the proband was suggested by overexpression and aberrant localization of RPA32 in tumor cells as compared with surrounding normal ductal tissue and by the presence of anti-RPA32 antibodies before the diagnosis. The prevalence of anti-RPA32 antibodies was significantly higher (P < 0.01) among breast cancer patients (87 of 801 patients) than among noncancer controls (0 of 65 controls). Similarly, anti-RPA32 antibodies were present in 4 of 39 patients with intraductal in situ carcinoma. No associations were found between anti-RPA antibodies and survival, occurrence of a second tumor, metastases, or antibodies to p53. Reactivity to RPA32 also was detected in sera from 3 of 47 patients with other cancers. CONCLUSIONS: In view of the central role of RPA in DNA replication, recombination, and repair, we suggest that autoimmunity to RPA32 may reflect molecular changes involved in the process of tumorigenesis. The finding of antibodies to RPA32 before diagnosis and their prevalence in in situ carcinoma suggest that they are potentially useful markers of early disease. PMID- 11895909 TI - Methylthioadenosine phosphorylase gene deletions are common in osteosarcoma. AB - PURPOSE: Methylthioadenosine phosphorylase (MTAP) is an enzyme essential in the salvage of cellular adenine and methionine synthesis. The MTAP gene is located in the 9p21 chromosomal region and its loss is frequently associated with deletion of the tumor suppressor genes p15(INK4b) and p16(INK4a). The aim of this study was to investigate the frequency of molecular alterations in MTAP in osteosarcoma. EXPERIMENTAL DESIGN: Samples from patients with high-grade osteosarcoma (n = 96) and three osteosarcoma cell lines (HOS, SaOS-2, and U2OS) were analyzed. Genomic DNA was analyzed for MTAP gene deletions by PCR, RNA expression was measured by semiquantitative reverse transcription-PCR, and the protein levels were measured by immunohistochemistry. RESULT: Deletion of at least one MTAP exon was found in 36 of 96 (37.5%) osteosarcoma patient samples and in one of the three cell lines (HOS). In all cases in which an MTAP gene deletion was observed, there was absence of detectable mRNA and protein. Furthermore, in four osteosarcoma patients, an MTAP deletion which was not evident at diagnosis was detected in subsequent tumor samples. CONCLUSIONS: The MTAP gene is commonly deleted in osteosarcoma patient samples, leading to an absence of mRNA and protein expression; these results indicate that inhibitors of de novo purine synthesis or methionine depletion may be effective as treatments for osteosarcoma patients whose tumors fail to express MTAP. PMID- 11895910 TI - Absence of HER2/neu gene expression in osteosarcoma and skeletal Ewing's sarcoma. AB - PURPOSE: Osteosarcoma (OS) and skeletal Ewing's sarcoma (EWS), the most common pediatric primary bone tumors, are aggressive malignancies with a tendency for early pulmonary metastasis. Advances in therapy have increased the overall 5-year survival to approximately 70%; however, patients who relapse often fail to respond to salvage therapy. Thus, more effective adjuvant chemotherapy is needed for these patients. Several reports have claimed expression of the HER2/neu (c erbB-2) gene in a high percentage of OSs and that its expression is a poor prognostic factor and have advocated monoclonal antibody therapy in those cases. EXPERIMENTAL DESIGN AND RESULTS: To validate the expression of HER2/neu in these tumors, archival cases of OS (n = 66) and EWS (n = 11) from 44 patients were assessed for HER2/neu gene expression by immunohistochemistry (DAKO rabbit polyclonal antibody A0485). Thirty-four cases (44%) demonstrated granular cytoplasmic staining, but none showed the distinct membranous staining characteristic of expression exhibited by breast carcinomas. To validate these immunohistochemical results, reverse transcription-PCR using RNA derived from archival material (n = 48) and several different primer pairs also failed to demonstrate the presence of amplifiable HER2/neu mRNA, although a known HER2/neu positive breast carcinoma showed amplifiable mRNA. CONCLUSIONS: Thus, in contrast to previous reports, our results demonstrate an absence of HER2/neu expression in OSs and EWSs. Our results show that HER2/neu is not expressed by these sarcomas, and we conclude that HER2/neu is therefore not an important prognostic factor and that anti-HER2/neu monoclonal antibody therapy is not appropriate for these patients. PMID- 11895911 TI - The feasibility of using fine needle aspiration from primary breast cancers for cDNA microarray analyses. AB - PURPOSE: Our aims in this pilot study were to determine whether fine needle aspirates (FNAs) provide a sufficient quantity of mRNA for cDNA microarray analysis, produce a set of quality control criteria to accept individual arrays, and determine whether gene expression profiles obtained from FNAs were representative of the source tumor. EXPERIMENTAL DESIGN: Twenty-seven women with breast cancer for treatment with primary surgery had a FNA before and at the time of surgery, and a portion of excised tumor was taken for array analysis. Control experiments were performed using two Ewing's sarcoma xenograft models. mRNA was extracted from the samples and hybridized with the reference (MCF7 cell line) on cDNA microarrays. Statistical methods were applied to identify acceptability criteria for the arrays. RESULTS: Statistical analyses demonstrated that an adequate array could be identified by calculating the SD of the log of fluorescence intensities from the arrays. Using this criterion, only 4 of the 27 patients (15%) had FNA samples suitable for array analysis. Gene expression profiles from the FNAs closely resembled that of the corresponding source tumors and were clearly distinguished from FNAs derived from the xenografts. CONCLUSIONS: SD is a useful quality index for the clinical application of cDNA microarrays. This "proof of principle" study demonstrates that FNAs from primary breast cancers can be used for microarray analysis, although without amplification, it is feasible in only a small proportion of patients. For this to be clinically useful, validated amplification techniques for FNA samples are probably required. PMID- 11895912 TI - Polymorphisms in genes encoding drugs and xenobiotic metabolizing enzymes, DNA repair enzymes, and response to treatment of childhood acute lymphoblastic leukemia. AB - PURPOSE: The most common childhood malignancy, acute lymphoblastic leukemia (ALL), remains the leading cause of cancer-related death in children because of resistant cases in which underlying predisposing factors are poorly understood. The interindividual variation in the activity of xenobiotic metabolizing enzymes that modify individual somatic mutation burden in the context of environmental exposure was shown to modify susceptibility to childhood ALL. Variable DNA repair capacity may further modulate induced DNA lesions. Similarly, differential capacity of ALL patients to process carcinogens and chemotherapeutic drugs could both modify an individual's risk of recurrent malignancy and response to therapy. EXPERIMENTAL DESIGN: We investigated the relationship between the risk of relapse in ALL patients and functional polymorphisms in genes encoding carcinogen metabolizing enzymes, including CYP1A1, CYP2D6, CYP2E1, MPO, GSTM1, GSTT1, GSTP1, NAT1, NAT2, NQO1, as well as DNA-repair enzymes hMLH1, hMSH3, XRCC1, XPF, and APE. Our study included 320 children with ALL, of which 68 relapsed or died because of this disease within 5 years of follow-up. RESULTS: Among children of the latter group, we found that carriers of CYP1A1*2A and NQO1*2 variants had worse disease prognosis according to Kaplan-Meier (P = 0.003) and Cox regression (P 150 nucleotides. The deduced proteins of 44 ORFs showed significant homologies to other proteins present in sequence databases, whereas 14 putative proteins did not. For 29 proteins, we could deduce a putative function. Most of these are related to the basic phage propagation cycle. The phi P27 genome represents a mosaic composed of genetic elements which are obviously derived from related and unrelated phages. We identified five short linker sequences of 22 to 151 bp in the phi P27 sequence which have also been detected in a couple of other lambdoid phages. These linkers are located between functional modules in the phage genome and are thought to play a role in genetic recombination. Although the overall DNA sequence of phi P27 is not highly related to other known phages, the data obtained demonstrate a typical lambdoid genome structure. PMID- 11895954 TI - Binding component of Clostridium perfringens iota-toxin induces endocytosis in Vero cells. AB - Clostridium perfringens iota-toxin is a binary toxin consisting of two individual proteins, the binding component (Ib) and the enzyme component (Ia). Wild-type Ib bound to Vero cells at 4 and 37 degrees C and formed oligomers at 37 degrees C but not at 4 degrees C. The Ib-induced K(+) release from the cells was dependent on the oligomer formation of Ib in the cells, but the oligomer formation did not induce rounding activity or cytotoxicity. After incubation of the cells with recombinant Ib (rIb) at 37 degrees C, the Ib oligomer in the cell became resistant to pronase treatment with time, but the Ib monomer was sensitive to the treatment. Furthermore, treatment of Vero cells with rIb in the presence of bafilomycin, methylamine, or ethylamine resulted in accumulation of the oligomer in the cells but had no effect on K(+) release. Moreover, incubation with Ib plus Ia in the presence of these agents caused no rounding in the cells. These observations suggest that Ib binds to Vero cells, itself oligomerizing to form ion-permeable channels and that the formation of oligomer then induces endocytosis. PMID- 11895956 TI - Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity. AB - Three enterotoxins from the Aeromonas hydrophila diarrheal isolate SSU have been molecularly characterized in our laboratory. One of these enterotoxins is cytotoxic in nature, whereas the other two are cytotonic enterotoxins, one of them heat labile and the other heat stable. Earlier, by developing an isogenic mutant, we demonstrated the role of a cytotoxic enterotoxin in causing systemic infection in mice. In the present study, we evaluated the role of these three enterotoxins in evoking diarrhea in a murine model by developing various combinations of enterotoxin gene-deficient mutants by marker-exchange mutagenesis. A total of six isogenic mutants were prepared in a cytotoxic enterotoxin gene (act)-positive or -negative background strain of A. hydrophila. We developed two single knockouts with truncation in either the heat-labile (alt) or the heat-stable (ast) cytotonic enterotoxin gene; three double knockouts with truncations of genes encoding (i) alt and ast, (ii) act and alt, and (iii) act and ast genes; and a triple-knockout mutant with truncation in all three genes, act, alt, and ast. The identity of these isogenic mutants developed by double crossover homologous recombination was confirmed by Southern blot analysis. Northern and Western blot analyses revealed that the expression of different enterotoxin genes in the mutants was correspondingly abrogated. We tested the biological activity of these mutants in a diet-restricted and antibiotic-treated mouse model with a ligated ileal loop assay. Our data indicated that all of these mutants had significantly reduced capacity to evoke fluid secretion compared to that of wild-type A. hydrophila; the triple-knockout mutant failed to induce any detectable level of fluid secretion. The biological activity of selected A. hydrophila mutants was restored after complementation. Taken together, we have established a role for three enterotoxins in A. hydrophila-induced gastroenteritis in a mouse model with the greatest contribution from the cytotoxic enterotoxin Act, followed by the Alt and Ast cytotonic enterotoxins. PMID- 11895955 TI - Yersinia enterocolitica as a vehicle for a naked DNA vaccine encoding Brucella abortus bacterioferritin or P39 antigen. AB - Brucella is a facultative intracellular parasite that causes brucellosis in animals and humans. The protective immune response against Brucella involves both humoral and cell-mediated immunity. In previous studies, we demonstrated that the T-dominant Brucella antigens bacterioferritin (BFR) and P39 administered either as CpG adjuvant recombinant proteins or as naked-DNA plasmids induced a specific Th1-biased immune response in mice. In order to improve the protection conferred by the BFR and P39 vaccines and to evaluate the additive role of antilipopolysaccharide (anti-LPS) antibodies, we used live attenuated Yersinia enterocolitica serotypes O:3 and O:9 as delivery vectors for naked-DNA plasmids encoding these BFR and P39 antigens. Following two intragastric immunizations in BALB/c mice, the Yersinia vectors harboring a DNA vaccine encoding BFR or P39 induced antigen-specific serum immunoglobulin and Th1-type responses (both lymphocyte proliferation and gamma interferon production) among splenocytes. Moreover, as expected, antibodies recognizing Brucella abortus 544 lipopolysaccharide were detected in O:9-immunized mice but not in O:3-treated animals. Animals immunized with O:9 organisms carrying pCI or with O:9 organisms alone were found to be significantly resistant to infection by B. abortus 544. Our data demonstrated that pCI plasmids encoding BFR or P39 and delivered with live attenuated strains of Yersinia O:3 or O:9 can trigger Th1-type responses. The fact than only O:9 vectors induced a highly significant protective immunity against B. abortus 544 infection pointed out the crucial role of anti-LPS antibodies in protection. The best protection was conferred by a serotype O:9 strain carrying pCIP39, confirming the importance of the P39 T-cell antigen in this mechanism. PMID- 11895957 TI - In vivo clearance of an intracellular bacterium, Francisella tularensis LVS, is dependent on the p40 subunit of interleukin-12 (IL-12) but not on IL-12 p70. AB - To determine the role of interleukin-12 (IL-12) in primary and secondary immunity to a model intracellular bacterium, we have comprehensively evaluated infection with Francisella tularensis LVS in three murine models of IL-12 deficiency. Mice lacking the p40 protein of IL-12 (p40 knockout [KO] mice) and mice treated in vivo with neutralizing anti-IL-12 antibodies survived large doses of primary and secondary LVS infection but never cleared bacteria and exhibited a chronic infection. In dramatic contrast, mice lacking the p35 protein (p35 KO mice) of heterodimeric IL-12 readily survived large doses of primary sublethal LVS infection as well as maximal secondary lethal challenge, with only a slight delay in clearance of bacteria. LVS-immune wild-type (WT) lymphocytes produced large amounts of gamma interferon (IFN-gamma), but p35 KO and p40 KO lymphocytes produced much less; nonetheless, similar amounts of NO were found in all cultures containing immune lymphocytes, and all immune lymphocytes were equally capable of controlling intracellular growth of LVS in vitro. Purified CD4(+) and CD8(+) T cells from both WT and p40 KO mice controlled intracellular growth, even though T cells from WT mice produced much more IFN-gamma than those from p40 KO mice, and p40 KO T cells did not adopt a Th2 phenotype. Thus, while IL-12 p70 stimulation of IFN-gamma production may be important for bacteriostasis, IL-12 p70 is not necessary for appropriate development of LVS-immune T cells that are capable of controlling intracellular bacterial growth and for clearance of primary or secondary LVS infection. Instead, an additional mechanism dependent on the IL-12 p40 protein, either alone or in another complex such as the newly discovered heterodimer IL-23, appears to be responsible for actual clearance of this intracellular bacterium. PMID- 11895958 TI - Coexpression of interleukin-12 chains by a self-splicing vector increases the protective cellular immune response of DNA and Mycobacterium bovis BCG vaccines against Mycobacterium tuberculosis. AB - More effective vaccines against Mycobacterium tuberculosis may contribute to the control of this major human pathogen. DNA vaccines encoding single mycobacterial proteins stimulate antimycobacterial T-cell responses and induce partial protection against M. tuberculosis in animal models. The protective efficacy of these vaccines encoding a single antigen, however, has been less than that afforded by the current vaccine, Mycobacterium bovis bacillus Calmette-Guerin (BCG). The heterodimeric cytokine interleukin-12 (IL-12) potentiates the induction and maintenance of the type 1 helper T-cell response. We have developed a novel self-splicing vector based on the 2A protein of foot-and-mouth disease virus that permits the coordinate expression of both chains of IL-12 (p2AIL12). Coimmunization with this vector and DNA expressing M. tuberculosis antigen 85B or MPT64 enhanced the specific lymphocyte proliferative response and increased the frequency of specific gamma interferon-secreting T cells against the whole protein and a defined CD8(+) T-cell epitope on MPT64. Further, coimmunizing with p2AIL12 significantly increased the protective efficacy of DNA-85 in the lung against an aerosol challenge with M. tuberculosis to the level achieved with BCG. Therefore, codelivery of an IL-12-secreting plasmid may be a potent strategy for enhancing the protective efficacy of vaccines against M. tuberculosis. PMID- 11895959 TI - Preexisting inflammation due to Mycobacterium bovis BCG infection differentially modulates T-cell priming against a replicating or nonreplicating immunogen. AB - Induction of T-cell memory by vaccination ensures long-term protection against pathogens. We determined whether on-going inflammatory responses during vaccination influenced T-cell priming. A preexposure of mice to Mycobacterium bovis BCG impaired their subsequent ability to prime T cells against Listeria monocytogenes. This was characterized by a decrease in L. monocytogenes-specific gamma interferon (IFN-gamma)-secreting CD4(+) and CD8(+) T cells. The intensity of T-cell priming towards L. monocytogenes depended on the extent of L. monocytogenes expansion, and a cessation of this expansion caused by M. bovis BCG induced inflammation resulted in impairment in T-cell priming. A challenge of M. bovis BCG-infected mice with a higher L. monocytogenes dose increased L. monocytogenes survival and restored T-cell priming towards L. monocytogenes. Impairment in T-cell priming towards L. monocytogenes due to M. bovis BCG-induced inflammation resulted in a compromised protective efficacy in the long term after mice were rechallenged with L. monocytogenes. Preexisting inflammation selectively impaired T-cell priming for replicating immunogens as CD8(+) T-cell response to ovalbumin administered as an inert antigen (ovalbumin-archaeosomes) was enhanced by M. bovis BCG preimmunization, whereas priming towards ovalbumin administered as a live immunogen (L. monocytogenes-ovalbumin) was impaired. Thus, depending on the nature of the immunogen, the presence of prior inflammatory responses may either impede or boost vaccine efficacy. PMID- 11895960 TI - Randomized, controlled human challenge study of the safety, immunogenicity, and protective efficacy of a single dose of Peru-15, a live attenuated oral cholera vaccine. AB - Peru-15 is a live attenuated oral vaccine derived from a Vibrio cholerae O1 El Tor Inaba strain by a series of deletions and modifications, including deletion of the entire CT genetic element. Peru-15 is also a stable, motility-defective strain and is unable to recombine with homologous DNA. We wished to determine whether a single oral dose of Peru-15 was safe and immunogenic and whether it would provide significant protection against moderate and severe diarrhea in a randomized, double-blind, placebo-controlled human volunteer cholera challenge model. A total of 59 volunteers were randomly allocated to groups to receive either 2 x 10(8) CFU of reconstituted, lyophilized Peru-15 vaccine diluted in CeraVacx buffer or placebo (CeraVacx buffer alone). Approximately 3 months after vaccination, 36 of these volunteers were challenged with approximately 10(5) CFU of virulent V. cholerae O1 El Tor Inaba strain N16961, prepared from a standardized frozen inoculum. Among vaccinees, 98% showed at least a fourfold increase in vibriocidal antibody titers. After challenge, 5 (42%) of the 12 placebo recipients and none (0%) of the 24 vaccinees had moderate or severe diarrhea (> or = 3,000 g of diarrheal stool) (P = 0.002; protective efficacy, 100%; lower one-sided 95% confidence limit, 75%). A total of 7 (58%) of the 12 placebo recipients and 1 (4%) of the 24 vaccinees had any diarrhea (P < 0.001; protective efficacy, 93%; lower one-sided 95% confidence limit, 62%). The total number of diarrheal stools, weight of diarrheal stools, incidence of fever, and peak stool V. cholerae excretion among vaccinees were all significantly lower than in placebo recipients. Peru-15 is a well-tolerated and immunogenic oral cholera vaccine that affords protective efficacy against life-threatening cholera diarrhea in a human volunteer challenge model. This vaccine may therefore be a safe and effective tool to prevent cholera in travelers and is a strong candidate for further evaluation to prevent cholera in an area where cholera is endemic. PMID- 11895961 TI - Influence of recombination and niche separation on the population genetic structure of the pathogen Streptococcus pyogenes. AB - The throat and skin of the human host are the principal reservoirs for the bacterial pathogen Streptococcus pyogenes. The emm locus encodes structurally heterogeneous surface fibrils that play numerous roles in virulence, depending on the strain. Isolates harboring the emm pattern A-C marker exhibit a strong tendency to cause throat infection, whereas emm pattern D strains are usually recovered from impetigo lesions; as a group, emm pattern E organisms fail to display obvious tissue tropisms. The peak incidence for streptococcal pharyngitis and impetigo varies with season and locale, leading to wide spatial and temporal distances between throat and skin strains. To assess any impact of niche separation on genetic variation, the extent of recombinational exchange between emm pattern A-C, D, and E subpopulations was evaluated. Analysis of nucleotide sequence data for internal portions of seven housekeeping loci from 212 isolates provides evidence of extensive recombination between strains belonging to different emm pattern subpopulations. Furthermore, no fixed nucleotide differences were found between emm pattern A-C and D strains. Thus, despite some niche separation created by distinct epidemiological trends and innate tissue tropisms there is little evidence for neutral gene divergence between throat and skin strains. Maintenance of a relationship between emm pattern and tissue tropism in the face of underlying recombination suggests that tissue tropism is associated with emm or a closely linked gene. PMID- 11895962 TI - Helicobacter pylori uses motility for initial colonization and to attain robust infection. AB - Helicobacter pylori has been shown to require flagella for infection of the stomach. To analyze whether flagella themselves or motility is needed by these pathogens, we constructed flagellated nonmotile mutants. This was accomplished by using both an insertion mutant and an in-frame deletion of the motB gene. In vitro, these mutants retain flagella (Fla(+)) but are nonmotile (Mot(-)). By using FVB/N mice, we found that these mutants had reduced ability to infect mice in comparison to that of their isogenic wild-type counterparts. When these mutants were coinfected with wild type, we were unable to detect any motB mutant. Finally, by analyzing the 50% infectious dose, we found that motility is needed for initial colonization of the stomach mucosa. These results support a model in which motility is used for the initial colonization of the stomach and also to attain full infection levels. PMID- 11895963 TI - Involvement of PhoP-PhoS homologs in Enterococcus faecalis virulence. AB - Eleven PhoP-PhoS homolog pairs were identified by searching the Enterococcus faecalis V583 genome sequence database at The Institute for Genomic Research with the Bacillus subtilis PhoP-PhoS sequences. Each pair appears to be a potential two-component system composed of a response regulator and a sensor kinase. Seven of the homologs were disrupted in E. faecalis strain OG1RF. TX10293, a mutant disrupted in one of these genes (etaR, the first gene of the gene pair designated etaRS), showed delayed killing and a higher 50% lethal dose in a mouse peritonitis model. The predicted EtaR protein sequence showed greatest similarity to LisR of Listeria monocytogenes (77%) and CsrR of Streptococcus pyogenes (70%); EtaS is 53% similar to LisK and 54% similar to CsrS. When grown in vitro, the TX10293 mutant was more sensitive to low pH (pH 3.4) and more resistant to high temperature (55 degrees C) than wild-type OG1RF. In conclusion, many potential two-component systems are identified for E. faecalis, one of which, EtaRS, was shown to be involved in stress response and virulence. PMID- 11895964 TI - Host immune response to Salmonella enterica serovar Typhimurium infection in mice derived from wild strains. AB - Studies of mouse models of endotoxemia and sepsis with gram-negative bacteria have shown that the host response is genetically controlled. Mice infected with the gram-negative bacterium Salmonella enterica serovar Typhimurium exhibit marked genetic differences in disease manifestation, and the wild-derived strain Mus musculus molossinus MOLF/Ei is extremely susceptible to S. enterica serovar Typhimurium. The kinetics of bacterial proliferation within the liver and the spleen and histological examination of tissue sections have suggested that MOLF/Ei mice do not succumb to infection because of overwhelming bacterial growth in the reticuloendothelial organs or massive tissue necrosis, as observed in other Salmonella-susceptible strains. MOLF/Ei mice respond normally to lipopolysaccharide (LPS) in vivo and in vitro, as determined by the production of tumor necrosis factor alpha and spleen cell mitogenesis. However, they have a unique cytokine profile in response to infection compared to that observed for other Salmonella-susceptible mice. There was increased expression of mRNA of the interleukin-1 alpha (IL-1 alpha) and IL-1 beta genes as the infection in the spleens and livers of MOLF/Ei mice progressed. Despite the fact that MOLF/Ei mice have the ability to respond to LPS and the fact that there are significant increases in IL-1 alpha and IL-1 beta mRNA, Nos2 in the spleen is not upregulated and nitrite production by spleen cells is reduced. At the central level, the inflammatory response is characterized by strong upregulation of the inhibitory factor kappa B alpha and Toll-like receptor 2 genes, two genes known to be regulated by LPS and IL-1 in the brain. The high levels of IL-1 expression in the spleens and livers of MOLF/Ei mice may have important implications for the activation of peripheral and central innate immune mechanisms. PMID- 11895965 TI - Evidence that the enterococcal polysaccharide antigen gene (epa) cluster is widespread in Enterococcus faecalis and influences resistance to phagocytic killing of E. faecalis. AB - In previous studies, we cloned a cluster of genes involved in polysaccharide biosynthesis (epa) from Enterococcus faecalis strain OG1RF and showed that this gene cluster mediated synthesis of a polysaccharide in Escherichia coli. Disruption of two open reading frames in the epa gene cluster of OG1RF generated two mutants, TX5179 and TX5180, which were attenuated in a mouse peritonitis model. In the current study, Western blotting was performed with serum from a patient with E. faecalis endocarditis and polysaccharide extracts from OG1RF and the mutants TX5179 and TX5180. OG1RF showed a smear in the high-molecular-weight region and discrete bands in the low-molecular-weight region, which were missing from the mutants; periodate treatment and carbohydrate staining confirmed the polysaccharide nature of this material. In a neutrophil killing assay using OG1RF absorbed normal human serum, the mutants TX5179 and TX5180, respectively, were 50 and 2.4 times more susceptible to killing than wild-type OG1RF (P < or = 0.01). With a fluorescence phagocytosis assay, 2.5 to 3 times more of the mutants were taken up by neutrophils than OG1RF (P < or = 0.001). Finally, with restriction digestion and hybridization under high-stringency conditions, the epa gene cluster of OG1RF (which is also present in the sequenced E. faecalis strain V583) was detected in 12 of 12 other clonally distinct E. faecalis strains tested: a similar polysaccharide pattern was detected for the 12 strains on Western blots using an E. faecalis endocarditis patient serum, and sera from four other patients with E. faecalis endocarditis all reacted with polysaccharide extracts of OG1RF. These results indicate that the epa gene cluster is widespread among E. faecalis and confers some protection against human host defenses. PMID- 11895966 TI - Phase I evaluation of delta virG Shigella sonnei live, attenuated, oral vaccine strain WRSS1 in healthy adults. AB - We conducted a phase I trial with healthy adults to evaluate WRSS1, a live, oral Delta virG Shigella sonnei vaccine candidate. In a double-blind, randomized, dose escalating fashion, inpatient volunteers received a single dose of either placebo (n = 7) or vaccine (n = 27) at 3 x 10(3) CFU (group 1), 3 x 10(4) CFU (group 2), 3 x 10(5) CFU (group 3), or 3 x 10(6) CFU (group 4). The vaccine was generally well tolerated, although a low-grade fever or mild diarrhea occurred in six (22%) of the vaccine recipients. WRSS1 was recovered from the stools of 50 to 100% of the vaccinees in each group. The geometric mean peak anti-lipopolysaccharide responses in groups 1 to 4, respectively, were 99, 39, 278, and 233 for immunoglobulin (IgA) antibody-secreting cell counts; 401, 201, 533, and 284 for serum reciprocal IgG titers; and 25, 3, 489, and 1,092 for fecal IgA reciprocal titers. Postvaccination increases in gamma interferon production in response to Shigella antigens occurred in some volunteers. We conclude that WRSS1 vaccine is remarkably immunogenic in doses ranging from 10(3) to 10(6) CFU but elicits clinical reactions that must be assessed in further volunteer trials. PMID- 11895967 TI - Mucosal or parenteral administration of microsphere-associated Bacillus anthracis protective antigen protects against anthrax infection in mice. AB - Existing licensed anthrax vaccines are administered parenterally and require multiple doses to induce protective immunity. This requires trained personnel and is not the optimum route for stimulating a mucosal immune response. Microencapsulation of vaccine antigens offers a number of advantages over traditional vaccine formulations, including stability without refrigeration and the potential for utilizing less invasive routes of administration. Recombinant protective antigen (rPA), the dominant antigen for protection against anthrax infection, was encapsulated in poly-L-lactide 100-kDa microspheres. Alternatively, rPA was loosely attached to the surfaces of microspheres by lyophilization. All of the microspheric formulations were administered to A/J mice with a two-dose schedule by either the intramuscular route, the intranasal route, or a combination of these two routes, and immunogenicity and protective efficacy were assessed. An intramuscular priming immunization followed by either an intramuscular or intranasal boost gave optimum anti-rPA immunoglobulin G titers. Despite differences in rPA-specific antibody titers, all immunized mice survived an injected challenge consisting of 10(3) median lethal doses of Bacillus anthracis STI spores. Immunization with microencapsulated and microsphere-associated formulations of rPA also protected against aerosol challenge with 30 median lethal doses of STI spores. These results show that rPA can be encapsulated and surface bound to polymeric microspheres without impairing its immunogenicity and also that mucosal or parenteral administration of microspheric formulations of rPA efficiently protects mice against both injected and aerosol challenges with B. anthracis spores. Microspheric formulations of rPA could represent the next generation of anthrax vaccines, which could require fewer doses because they are more potent, are less reactogenic than currently available human anthrax vaccines, and could be self-administered without injection. PMID- 11895968 TI - Construction of a tetR-integrated Salmonella enterica serovar Typhi CVD908 strain that tightly controls expression of the major merozoite surface protein of Plasmodium falciparum for applications in human Vaccine production. AB - Attenuated Salmonella strains are an attractive live vector for delivery of a foreign antigen to the human immune system. However, the problem with this vector lies with plasmid segregation and the low level of expression of the foreign gene in vivo when constitutive expression is employed, leading to a diminished immune response. We have established inducible expressions of foreign genes in the Salmonella enterica serovar Typhi CVD908 vaccine strain using the tetracycline response regulatory promoter. To set up this system, a tetracycline repressor (tetR) was integrated into a defined Delta aroC locus of the chromosome via suicide plasmid pJG12/tetR-neo. To remove the neo gene conferring kanamycin resistance from the locus, a cre expression vector under the control of the tetracycline response promoter was transformed into the clone; expression of the Cre recombinase excised the neo gene and generated the end strain CVD908-tetR. Expression of the luciferase reporter gene in this strain is dependent on the presence of tetracycline in the medium and can be regulated up to 4,773-fold. Moreover, the tightly controlled expression of major merozoite surface protein 1 (MSP1) and parts of Plasmodium falciparum was achieved, and the product yield was increased when the inducible expression system was employed. Inoculation of bacteria harboring plasmid pZE11/MSP1(42) in mice produced the protein in liver and spleen controlled by the inducer. The persistence of the plasmid-carrying bacteria in mice was determined. Peak colonization of both liver and spleen was detected on the third day postinoculation and was followed by a decline in growth curves. After 14 days postinfection, the majority of the bacteria (>90%) recovered from the liver and spleen of the mice retained the plasmid when expression was induced; this clearly indicated that stability of the expression vector in vivo was improved by inducible expression. Establishment of the regulatory system in the vaccine strain may broaden the range of its use by enhancing plasmid stability and expression levels in vivo. Moreover, the availability of the vaccine strain inducibly expressing the entire MSP1 provides possibilities for examining its immunogenicity, particularly the cellular response in animal models. PMID- 11895969 TI - Immunodominant epitopes in Babesia bovis rhoptry-associated protein 1 that elicit memory CD4(+)-T-lymphocyte responses in B. bovis-immune individuals are located in the amino-terminal domain. AB - Babesia bovis rhoptry-associated protein 1 (RAP-1), which confers partial protection against B. bovis challenge, is recognized by antibodies and T lymphocytes from cattle that have recovered from infection and are immune to subsequent challenge. RAP-1 is a 60-kDa protein with an N-terminal (NT) region that contains four cysteine residues conserved among all Babesia RAP-1 family members and a C-terminal (CT) region that contains multiple, degenerate, tandem 23-amino-acid (aa) repeats. To define the location of CD4(+)-T-cell epitopes for vaccine development using a recombinant protein or minigene construct, a series of truncated recombinant RAP-1 proteins and peptides were tested for stimulation of T-cell lines derived from B. bovis-immune cattle. CD4(+)-T-cell lines from three B. bovis-immune cattle with different DRB3 haplotypes responded to the NT region of RAP-1, whereas T cells from only one animal responded weakly to the CT region. T-cell lines from the three individuals recognized two to six NT-region peptides spanning aa 134 to 316 and representing at least four dominant epitopes. Using RAP-1-specific CD4(+)-T-cell clones, two NT-region epitopes, EYLVNKVLYMATMNYKT (aa 187 to 203) and EAPWYKRWIKKFR (aa 295 to 307), and one CT region repeat epitope, FREAPQATKHFL, which is present twice at aa positions 391 to 402 and 414 to 425, were identified. Several peptides representing degenerate repeats of the agonist CT-region peptide FREAPQATKHFL neither stimulated responses of T-cell clones specific for this peptide nor inhibited responses to the agonist peptide. Upon stimulation with specific antigen, T-cell clones specific for NT or CT epitopes produced gamma interferon. The presence of T helper-cell epitopes in the NT domain of RAP-1, which is highly conserved among otherwise antigenically different strains of B. bovis, supports the inclusion of this region in vaccine constructs to be tested in cattle. PMID- 11895970 TI - Susceptibility to experimental cerebral malaria induced by Plasmodium berghei ANKA in inbred mouse strains recently derived from wild stock. AB - The neurological syndrome caused by Plasmodium berghei ANKA in rodents partially mimics the human disease. Several rodent models of cerebral malaria (CM) exist for the study of the mechanisms that cause the disease. However, since common laboratory mouse strains have limited gene pools, the role of their phenotypic variations causing CM is restricted. This constitutes an obstacle for efficient genetic analysis relating to the pathogenesis of malaria. Most common laboratory mouse strains are susceptible to CM, and the same major histocompatibility complex (MHC) haplotype may exhibit different levels of susceptibility. We analyzed the influence of the MHC haplotype on overcoming CM by using MHC congenic mice with C57BL/10 and C3H backgrounds. No correlation was found between MHC molecules and the development of CM. New wild-derived mouse strains with wide genetic polymorphisms were then used to find new models of resistance to CM. Six of the twelve strains tested were resistant to CM. For two of them, F(1) progeny and backcrosses performed with the reference strain C57BL/6 showed a high level of heterogeneity in the number and characteristics of the genetic factors associated with resistance to CM. PMID- 11895972 TI - Stachylysin may be a cause of hemorrhaging in humans exposed to Stachybotrys chartarum. AB - Stachybotrys chartarum is a toxigenic fungus that has been associated with human health concerns such as nasal bleeding in adults and pulmonary hemosiderosis (PH) in infants. Seven of eight strains of S. chartarum isolated from homes of infants with PH in Cleveland, Ohio, and the strain from the lung of an infant with PH in Texas produced stachylysin in tryptic soy broth (TSB), whereas only one out of eight strains isolated from control homes produced stachylysin. However, all strains produced stachylysin when grown on TSB with 0.7% sheep's blood. When stachylysin was injected into Lumbricus terrestis, the erythrocruorin hemoglobin (absorbance peaks at 280 and 415 nm) was released, resulting in a lethal effect. These results support the hypothesis that stachylysin may be one agent responsible for hemorrhaging in humans. PMID- 11895971 TI - Lipoteichoic acids from Lactobacillus johnsonii strain La1 and Lactobacillus acidophilus strain La10 antagonize the responsiveness of human intestinal epithelial HT29 cells to lipopolysaccharide and gram-negative bacteria. AB - Intestinal epithelial cells (IECs) respond to lipopolysaccharide (LPS) from gram negative bacteria in the presence of the soluble form of CD14 (sCD14), a major endotoxin receptor. Since sCD14 is also known to interact with gram-positive bacteria and their components, we looked at whether sCD14 could mediate their effects on human IECs. To this end, we examined the production of proinflammatory cytokines following exposure of the IECs to specific gram-positive bacteria or their lipoteichoic acids (LTAs) in the absence and presence of human milk as a source of sCD14. In contrast to LPS from Escherichia coli or Salmonella enteritidis, neither the gram-positive bacteria Lactobacillus johnsonii strain La1 and Lactobacillus acidophilus strain La10 nor their LTAs stimulated IECs, even in the presence of sCD14. However, both LTAs inhibited the sCD14-mediated LPS responsiveness of IECs. We have previously hypothesized that sCD14 in human milk is a means by which the neonate gauges the bacterial load in the intestinal lumen and liberates protective proinflammatory cytokines from IECs. The present observations suggest that gram-positive organisms, via their LTAs, temper this response and prevent an exaggerated inflammatory response. PMID- 11895973 TI - Mice lacking T and B lymphocytes develop transient colitis and crypt hyperplasia yet suffer impaired bacterial clearance during Citrobacter rodentium infection. AB - The bacterial pathogen Citrobacter rodentium belongs to a family of gastrointestinal pathogens that includes enteropathogenic and enterohemorrhagic Escherichia coli and is the causative agent of transmissible colonic hyperplasia in mice. The molecular mechanisms used by these pathogens to colonize host epithelial surfaces and form attaching and effacing (A/E) lesions have undergone intense study. In contrast, little is known about the host's immune response to these infections and its importance in tissue pathology and bacterial clearance. To address these issues, wild-type mice and mice lacking T and B lymphocytes (RAG1 knockout [KO]) were infected with C. rodentium. By day 10 postinfection (p.i.), both wild-type and RAG1 KO mice developed colitis and crypt hyperplasia, and these responses became more exaggerated in wild-type mice over the next 2 weeks, as they cleared the infection. By day 24 p.i., bacterial clearance was complete, and the colitis had subsided; however, crypt heights remained increased. In contrast, inflammatory and crypt hyperplastic responses in the RAG1 KO mice were transient, subsiding after 2 weeks. By day 24 p.i., RAG1 KO mice showed no signs of bacterial clearance and infection was often fatal. Surprisingly, despite remaining heavily infected, tissues from RAG1 KO mice surviving the acute colitis showed few signs of disease. These results thus emphasize the important contribution of the host immune response during infection by A/E bacterial pathogens. While T and/or B lymphocytes are essential for host defense against C. rodentium, they also mediate much of the tissue pathology and disease symptoms that occur during infection. PMID- 11895974 TI - Local role for tumor necrosis factor alpha in the pulmonary inflammatory response to Mycobacterium tuberculosis infection. AB - The local intrapulmonary role of tumor necrosis factor alpha (TNF-alpha) in a protective host response during acute and chronic infection with Mycobacterium tuberculosis is incompletely understood. To directly assess its role in the intrapulmonary immune response, we compared the responses of transgenic mice with a local pulmonary blockade of TNF-alpha (SPCTNFRIIFc mice) to mice with globally inhibited TNF-alpha (TNFRKO mice) and mice with normal immune systems (control mice). Consistent with previous reports, 100% of TNFRKO mice died by 28 days after aerosol infection, and these mice had markedly increased numbers of bacteria and widespread tissue necrosis in their lungs compared to controls. The median survival time of the SPCTNFRIIFc mice was 142 days, and 75% died by 180 days. Even though the numbers of bacteria in the lungs of the SPCTNFRIIFc mice were marginally increased compared to controls, these mice had a persistent neutrophilic inflammatory response and increased expression of proinflammatory cytokines (interleukin-1 alpha/beta [IL-1 alpha/beta], IL-18, gamma interferon, IL-6, and macrophage migration inhibitory factor) and chemokines (eotaxin, macrophage inflammatory protein 1 alpha/beta, gamma interferon-inducible protein 10, macrophage chemotaxic protein 1, and TCA-3) in their lungs. These studies with the SPCTNFRIIFc mice provide direct evidence for the local importance of TNF alpha in the proper regulation of host defense to M. tuberculosis. The studies also suggest that when the local actions of TNF-alpha are selectively impaired in the lungs, tissue destruction and death ensue, at least in part, due to persistent expression of proinflammatory mediators that would normally be downregulated. PMID- 11895975 TI - Role of gamma interferon in chemokine expression in the ileum of mice and in a murine intestinal epithelial cell line after Cryptosporidium parvum infection. AB - Cryptosporidium parvum is a protozoan parasite that infects intestinal epithelial cells and induces inflammation of the intestine. To better understand the inflammatory process occurring during cryptosporidiosis, we investigated in this study the kinetics of chemokine expression in the mucosa of mice by quantitative reverse transcription-PCR. Our results demonstrate that among the chemokine mRNAs studied, gamma interferon (IFN-gamma)-inducible protein 10 (IP-10), monokine induced by IFN-gamma (MIG), i-TAC, lymphotactin, macrophage inflammatory protein 1 beta (MIP-1 beta), and RANTES mRNAs were strongly up-regulated in infected neonate mice, which correlated with the immunofluorescence staining results showing T-cell and macrophage infiltration in the mucosa. Our in vitro data showed that intestinal epithelial cells infected by C. parvum or stimulated by the proinflammatory cytokines (IFN-gamma, interleukin-1 beta, and tumor necrosis factor alpha) produce a pattern of chemokine secretion similar to that observed in vivo, suggesting that these cells may take part in the initial production of chemokines. In order to identify the chemokines responsible for the recruitment of the inflammatory cells leading to a protective immune response, we compared the patterns of chemokine expression in a healing neonate mouse model and a nonhealing IFN-gamma knockout (GKO) mouse model of cryptosporidiosis. In the absence of IFN-gamma, the chemokine response was altered for IP-10, MIG, i-TAC, RANTES, and MIP-1 beta mRNAs, while the three ELR C-X-C chemokine mRNAs studied (lipopolysaccharide-induced C-X-C chemokine, MIP-2 alpha, and KC mRNAs) were strongly overexpressed. These results are consistent with the neutrophil recruitment observed in the lamina propria of GKO mice at day 9 postinfection but are not consistent with the hypothesis that these cells play an important role in the resolution of the infection. On the contrary, the altered response of chemokines responsible for the recruitment of macrophages and T cells in GKO mice suggests that these two populations may be critical in the development of a protective immune response. PMID- 11895976 TI - Mycobacterial antigens exacerbate disease manifestations in Mycobacterium tuberculosis-infected mice. AB - To control tuberculosis worldwide, the burden of adult pulmonary disease must be reduced. Although widely used, Mycobacterium bovis BCG vaccination given at birth does not protect against adult pulmonary disease. Therefore, postexposure vaccination of adults with mycobacterial antigens is being considered. We examined the effect of various mycobacterial antigens on mice with prior M. tuberculosis infection. Subcutaneous administration of live or heat-treated BCG with or without lipid adjuvants to infected mice induced increased antigen specific T-cell proliferation but did not reduce the bacterial load in the lungs and caused larger lung granulomas. Similarly, additional mycobacterial antigen delivered directly to the lungs by aerosol infection with viable M. tuberculosis mixed with heat-killed Mycobacterium tuberculosis (1:1) also did not reduce the bacillary load but caused increased expression of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6), which was associated with larger granulomas in the lungs. When M. tuberculosis-infected mice were treated with recombinant BCG that secreted cytokines shown to reduce disease in a preinfection vaccine model, the BCG secreting TNF-alpha, and to a lesser extent, IL-2 and gamma interferon (IFN-gamma), caused a significant increase in granuloma size in the lungs. Moreover, treatment of M. tuberculosis-infected mice with recombinant murine TNF-alpha resulted in increased inflammation in the lungs and accelerated mortality without affecting the bacillary load. Taken together, these studies suggest that administration of mycobacterial antigens to mice with prior M. tuberculosis infection leads to immune activation that may exacerbate lung pathology via TNF-alpha-induced inflammation without reducing the bacillary load. PMID- 11895978 TI - Upregulation of thymosin beta-10 by Mycobacterium bovis infection of bovine macrophages is associated with apoptosis. AB - Bovine macrophages underwent apoptosis as a result of infection with a Mycobacterium bovis field strain. Macrophages infected with a multiplicity of infection (MOI) of 25:1 developed chromatin condensation and DNA fragmentation at 4 h and 8 h, respectively, whereas changes in chromatin condensation induced by MOIs of 10:1 and 1:1 required more time and had a reduced number of apoptotic cells. Not only infected macrophages underwent apoptosis, but also uninfected bystander macrophages became apoptotic. Increased differential expression of thymosin beta-10 was identified in M. bovis-infected bovine macrophages by differential display reverse transcriptase PCR. Phagocytosis of latex beads had no effect on the expression of thymosin beta-10, whereas bacterial suspensions upregulated thymosin beta-10 expression, suggesting that M. bovis or mycobacterial products are essential in the process. Heat-inactivated M. bovis induced a slight increase in thymosin beta-10 mRNA, whereas live virulent and attenuated M. bovis organisms increased the gene expression almost twofold. A mouse macrophage cell line (RAW 264.7) overexpressing the bovine thymosin beta-10 transgene had spontaneous apoptosis at a higher rate (66.5%) than parental cells (4.7%) or RAW cells harboring the empty vector (22.8%). The apoptotic rates of the overexpressing cells were significantly higher when compared with both the empty vector transfected (P < 0.01) and parental cells (P < 0.001). Our evidence suggests that upregulation of thymosin beta-10 in M. bovis-infected macrophages is linked with increased cell death due to apoptosis. PMID- 11895977 TI - Specific entry of Helicobacter pylori into cultured gastric epithelial cells via a zipper-like mechanism. AB - Although Helicobacter pylori has generally been considered an extracellular pathogen, a number of in vitro infection experiments and biopsy examinations have shown that it is capable of occasionally entering mammalian host cells. Here, we characterized this entry process by using AGS cells as a host cell model. In gentamicin protection-invasion assays, the number of H. pylori colonies recovered was lower than that for Salmonella enterica serovar Typhimurium X22, Escherichia coli expressing InvA, and Yersinia enterocolitica YO:9 grown at 25 degrees C but higher than that for Neisseria gonorrhoeae VP1 and Y. enterocolitica YO:9 grown at 37 degrees C. At the ultrastructural level, the entry process was observed to occur via a zipper-like mechanism. Internalized H. pylori was bound in tight LAMP 1-containing vacuoles in close association with condensed filamentous actin and tyrosine phosphorylation signals. Wortmannin, a potent inhibitor of phosphatidylinositol 3-kinase, and calphostin C, an inhibitor of protein kinase C, both inhibited the entry of H. pylori in a sensitive and dose-dependent manner; however, the level of entry was enhanced by sodium vanadate, an inhibitor of tyrosine phosphatases and ATPases. Furthermore, the cytokine tumor necrosis factor alpha antagonized the entry of H. pylori into AGS cells. Collectively, these results demonstrate that the entry of H. pylori into AGS cells occurs via a zipper-like mechanism which involves various host signal transduction events. PMID- 11895979 TI - Characterization and transcriptional analysis of gene clusters for a type IV secretion machinery in human granulocytic and monocytic ehrlichiosis agents. AB - Anaplasma (Ehrlichia) phagocytophila and Ehrlichia chaffeensis, the etiologic agents of granulocytic and monocytic ehrlichioses, respectively, are obligatory intracellular bacteria that cause febrile systemic illness in humans. We identified and characterized clusters of genes for a type IV secretion machinery in these two bacteria, and analyzed their gene expression in cell culture and mammalian hosts. Eight virB and virD genes were found in each bacterial genome, and all of the genes were transcribed in cell culture. Although the gene order and orientation were similar to those found in other bacteria, the eight virB and virD genes were clustered at two separate loci in each genome. Five of the genes (virB8, virB9, virB10, virB11, and virD4) were located downstream from a ribA gene. These five genes in both A. phagocytophila and E. chaffeensis were polycistronically transcribed and controlled through at least two tandem promoters located upstream of the virB8 gene in human leukemia cell lines. The virB9 gene of A. phagocytophila was transcriptionally active in peripheral blood leukocytes from human ehrlichiosis patients and experimentally infected animals. Three of the remaining genes (virB3, virB4, and virB6) of both A. phagocytophila and E. chaffeensis were arranged downstream from a sodB gene and cotranscribed with the sodB gene through one or more sodB promoters in human leukocytes. This suggests that transcription of the three virB genes in these two Anaplasma and Ehrlichia spp. is regulated by factors that influence the sodB gene expression. This unique regulation of gene expression for the type IV secretion system may be associated with intracellular survival and replication of Anaplasma and Ehrlichia spp. in granulocytes or monocytes. PMID- 11895981 TI - Early enhanced Th1 response after Leishmania amazonensis infection of C57BL/6 interleukin-10-deficient mice does not lead to resolution of infection. AB - C3H and C57BL/6 mice are resistant to Leishmania major but develop chronic lesions with persistent parasite loads when they are infected with Leishmania amazonensis. These lesions develop in the absence of interleukin-4 (IL-4), indicating that susceptibility to this parasite is not a result of development of a Th2 response. Expression of the cytokine IL-10 during infection could account for the lack of IL-12 expression and poor cell-mediated immunity towards the parasite. Therefore, we tested the hypothesis that IL-10 plays a central role in downmodulating the Th1 response after L. amazonensis infection. Infection of C57BL/6 IL-10-deficient mice indicated that in the absence of IL-10 there was early enhancement of a Th1 response, which was downregulated during the more chronic stage of infection. In addition, although there were 1- to 2-log reductions in the parasite loads within the lesions, the parasites continued to persist, and they were associated with chronic lesions whose size was similar to that of the control lesions. These experiments indicated that L. amazonensis resistance to killing in vivo is only partially dependent on expression of host IL-10. However, IL-10-deficient mice had an enhanced delayed-type hypersensitivity response during the chronic phase of infection, indicating that there were Th1 type effector cells in vivo at this late stage of infection. These results indicate that although IL-10 plays a role in limiting the Th1 response during the acute infection phase, other immunomodulatory factors are responsible for limiting the Th1 response during the chronic phase. PMID- 11895980 TI - Clonal polymorphism of Borrelia burgdorferi strain B31 MI: implications for mutagenesis in an infectious strain background. AB - A major obstacle to studying the functions of particular gene products in the mouse-tick infectious cycle of Borrelia burgdorferi has been an inability to knock out genes in pathogenic strains. Here, we investigated conditions for site directed mutagenesis in B31 MI, the low-passage-number, infectious B. burgdorferi strain whose genome was sequenced. We inactivated several plasmid and chromosomal genes in B31 MI and determined that clones carrying these mutations were not infectious for mice. However, we found extensive heterogeneity among clones and mutants derived from B31 MI based on colony phenotype, growth rate, plasmid content, protein profile, and transformability. Significantly, several B31 MI clones that were not subjected to mutagenesis but that lacked particular plasmids also exhibited defects at various stages in the infectious cycle. Therefore, the high degree of clonal polymorphism within B31 MI complicates the assessment of the contributions of individual genes to the observed phenotypes of the mutants. Our results indicate that B31 MI is not an appropriate strain background for genetic studies in infectious B. burgdorferi, and a well-defined isogenic clone is a prerequisite for targeted mutagenesis. To this end, we derived several wild type clones from B31 MI that were infectious for mice, and gene inactivation was successful in one of these clones. Due to the instability of the genome with in vitro propagation, careful monitoring of plasmid content of derived mutants and complementation of inactivated genes will be crucial components of genetic studies with this pathogen. PMID- 11895982 TI - Vaccination of guinea pigs with DNA encoding the mycobacterial antigen MPB83 influences pulmonary pathology but not hematogenous spread following aerogenic infection with Mycobacterium bovis. AB - Protection of cattle against bovine tuberculosis by vaccination could be an important control strategy in countries where there is persistent Mycobacterium bovis infection in wildlife and in developing countries where it is not economical to implement a tuberculin test and slaughter control program. The main aim of such a vaccination strategy would be to reduce transmission of infection by reducing the lung pathology caused by infection and preventing seeding of the organism to organs from which M. bovis could be excreted. Recent reports of successful DNA vaccination against Mycobacterium tuberculosis in small-animal models have suggested that DNA vaccines act by reducing lung pathology without sensitizing animals to tuberculin testing. We therefore evaluated the ability of vaccines consisting of DNA encoding the mycobacterial antigens MPB83 and 85A to reduce lung pathology and prevent hematogenous spread in guinea pigs challenged with a low dose of aerosolized M. bovis. Vaccination with MPB83 DNA reduced the severity of pulmonary lesions, as assessed by histopathology, and resembled M. bovis BCG vaccination in this respect. However, unlike BCG vaccination, MPB83 DNA vaccination did not protect challenged guinea pigs from hematogenous spread of organisms to the spleen. In contrast, vaccination with antigen 85A DNA, a promising DNA vaccine for human tuberculosis, had no measurable protective effect against infection with M. bovis. PMID- 11895983 TI - Antibodies against a synthetic peptide of SagA neutralize the cytolytic activity of streptolysin S from group A streptococci. AB - Virtually all group A streptococci (GAS) produce streptolysin S (SLS), a cytolytic toxin that is responsible for the beta-hemolysis surrounding colonies of the organisms grown on blood agar. SLS is an important virulence determinant of GAS, and recent studies have identified a nine-gene locus that is responsible for synthesis and transport of the toxin. SLS is not immunogenic; thus, no neutralizing antibodies are evoked during the course of natural infection. In the present study, we show that a synthetic peptide containing amino acid residues 10 to 30 of the putative SLS (SagA) propeptide [SLS(10-30)] coupled to keyhole limpet hemocyanin evoked antibodies in rabbits that completely neutralized the hemolytic activity of the toxin in vitro. Inhibition of hemolysis was reversed by preincubation of the immune serum with soluble, unconjugated peptide, indicating the specificity of the antibodies. In addition, antibodies that were affinity purified over an SLS(10-30) peptide column completely inhibited SLS-mediated hemolysis. The SLS(10-30) antisera did not opsonize group A streptococci; however, when combined with type-specific M protein antisera, the SLS antibodies significantly enhanced phagocytosis mediated by M protein antibodies. Thus, we have shown for the first time that it is possible to raise neutralizing antibodies against one of the most potent bacterial cytolytic toxins known. Our data also provide convincing evidence that the sagA gene actually encodes the SLS peptide of GAS. The synthetic peptide may prove to be an important component of vaccines designed to prevent GAS infections. PMID- 11895984 TI - Immunogenicity of a 26-valent group A streptococcal vaccine. AB - A multivalent vaccine containing amino-terminal M protein fragments from 26 different serotypes of group A streptococci was constructed by recombinant techniques. The vaccine consisted of four different recombinant proteins that were formulated with alum to contain 400 microg of protein per dose. Rabbits were immunized via the intramuscular route at 0, 4, and 16 weeks. Immune sera were assayed for the presence of type-specific antibodies against the individual recombinant M peptides by enzyme-linked immunosorbent assay and for opsonic antibodies by in vitro opsonization tests and indirect bactericidal tests. The 26 valent vaccine was highly immunogenic and elicited fourfold or greater increases in antibody levels against 25 of the 26 serotypes represented in the vaccine. The immune sera were broadly opsonic and were bactericidal against the majority of the 26 different serotypes. Importantly, none of the immune sera cross-reacted with human tissues. Our results indicate that type-specific, protective M protein epitopes can be incorporated into complex, multivalent vaccines designed to elicit broadly protective opsonic antibodies in the absence of tissue-cross reactive antibodies. PMID- 11895985 TI - Identification of a transcytosis epitope on staphylococcal enterotoxins. AB - Staphylococcal enterotoxins (SE) are exoproteins produced by Staphylococcus aureus that act as superantigens and have been implicated as a leading cause of food-borne disease and toxic shock. Little is known about how these molecules penetrate the gut lining and gain access to both local and systemic immune tissues. To model movement in vitro of staphylococcal enterotoxins, we have employed a monolayer system composed of crypt-like human colonic T-84 cells. SEB and SEA showed comparable dose-dependent transcytosis in vitro, while toxic shock syndrome toxin (TSST-1) exhibited increased movement at lower doses. Synthetic peptides corresponding to specific regions of the SEB molecule were tested in vitro to identify the domain of the protein involved in the transcytosis of SE. A toxin peptide of particular interest contains the amino acid sequence KKKVTAQELD, which is highly conserved across all SE. At a toxin-to-peptide ratio of 1:10, movement of SEB across the monolayers was reduced by 85%. Antisera made against the SEB peptide recognized native SEB and also inhibited SEB transcytosis. Finally, the conserved 10-amino-acid peptide inhibited transcytosis of multiple staphylococcal enterotoxins, SEA, SEE, and TSST-1. These data demonstrate that this region of the staphylococcal enterotoxins plays a distinct role in toxin movement across epithelial cells. It has implications for the prevention of staphylococcal enterotoxin-mediated disease by design of a peptide vaccine that could reduce systemic exposure to oral or inhaled superantigens. Since the sequence identified is highly conserved, it allows for a single epitope blocking the transcytosis of multiple SE. PMID- 11895987 TI - Pseudomonas aeruginosa ExoT acts in vivo as a GTPase-activating protein for RhoA, Rac1, and Cdc42. AB - The Pseudomonas aeruginosa protein ExoT is a bacterial GTPase-activating protein (GAP) that has in vitro activity toward Rho, Rac, and Cdc42 GTPases. Expression of ExoT both inhibits the internalization of strain PA103 by macrophages and epithelial cells and is associated with morphological changes (cell rounding and detachment) of infected cells. We find that expression of ExoT leads to the loss of GTP-bound RhoA, Rac1, and Cdc42 in transfected HeLa cells, demonstrating that ExoT has GAP activity in vivo toward all three GTPases. GAP activity is absolutely dependent on the presence of arginine at position 149 but is not affected by whether ExoT is expressed in the absence or presence of other P. aeruginosa type III secreted proteins. We also demonstrate that expression of ExoT in epithelial cells is sufficient to cause stress fiber disassembly by means of ExoT's GAP activity toward RhoA. PMID- 11895988 TI - Cortactin is necessary for F-actin accumulation in pedestal structures induced by enteropathogenic Escherichia coli infection. AB - Cortactin and the translocated intimin receptor, Tir, interacted with each other in pedestal formation in HeLa cells infected with enteropathogenic Escherichia coli (EPEC). Cortactin is shown to be necessary for organizing actin pedestals in response to EPEC, based on the expression of green fluorescent protein-fused cortactin derivatives in HeLa cells. PMID- 11895986 TI - Balance of pro- and anti-inflammatory cytokines correlates with outcome of acute experimental Pseudomonas aeruginosa keratitis. AB - The purpose of this study was to elucidate the expression of pro- and anti inflammatory cytokines in mouse corneas infected with Pseudomonas aeruginosa. Three bacterial strains (invasive, cytotoxic, or CLARE [contact lens-induced acute red eye]) which have recently been shown to produce distinct patterns of corneal disease in the mouse were used. The left mouse (BALB/c) corneas were scarified and infected with 2 x 10(6) CFU of one of the three P. aeruginosa strains, while right eyes served as controls. Animals were examined at 1, 4, 8, 16, and 24 h with a slit lamp biomicroscope to grade the severity of infection. Following examination, eyes were collected and processed for histopathology, multiprobe RNase protection assay for cytokine mRNA, enzyme-linked immunosorbent assay to quantitate cytokine proteins, and myeloperoxidase activity to quantitate polymorphonuclear leukocytes. The kinetics of appearance and magnitude of expression of key cytokines varied significantly in the three different phenotypes of P. aeruginosa infection. The predominant cytokines expressed in response to all three phenotypes were interleukin-1 beta (IL-1 beta), IL-1Ra, and IL-6. In response to the invasive strain, which induced severe corneal inflammation, significantly lower ratios of IL-1Ra to IL-1 beta were present at all time points, whereas corneas challenged with the CLARE strain, which induced very mild inflammation, showed a high ratio of IL-1Ra to IL-1 beta. The outcome of infection in bacterial keratitis correlated with the relative induction of these pro- and anti-inflammatory cytokines, and exogenous administration of recombinant rIL-1Ra (rIL-1Ra) was able to reduce the disease severity significantly. These findings point to the therapeutic potential of rIL-1Ra protein in possible treatment strategies for bacterial keratitis. PMID- 11895989 TI - Recombinant tumor necrosis factor alpha does not inhibit the growth of African trypanosomes in axenic cultures. AB - Mice whose tumor necrosis factor alpha (TNF-alpha) genes were disrupted developed higher levels of parasitemia than wild-type mice following infection with Trypanosoma congolense IL1180 or T. brucei brucei GUTat3.1, confirming the results of earlier studies. To determine whether TNF-alpha directly affects the growth of these and other bloodstream forms of African trypanosomes, we studied the effects of recombinant mouse, human, and bovine TNF-alpha on the growth of two isolates of T. congolense, IL1180 and IL3338, and two isolates of T. brucei brucei, GUTat3.1 and ILTat1.1, under axenic culture conditions. The preparations of recombinant TNF-alpha used were biologically active as determined by their capacity to kill L929 cells. Of five recombinant TNF-alpha lots tested, one lot of mouse TNF-alpha inhibited the growth of both isolates of T. brucei brucei and one lot of bovine TNF-alpha inhibited the growth of T. brucei brucei ILTat1.1 but only at very high concentrations and without causing detectable killing of the parasites. The other lots of mouse recombinant TNF-alpha, as well as human TNF alpha, did not affect the growth of any of the test trypanosomes even at maximal concentrations that could be attained in the culture systems (3,000 to 15,000 U of TNF-alpha/ml of medium). These results suggest that exogenously added recombinant TNF-alpha generally does not inhibit the growth of African trypanosomes under the culture conditions we used. The impact of TNF-alpha on trypanosome parasitemia may be indirect, at least with respect to the four strains of trypanosomes reported here. PMID- 11895990 TI - NK T cells are a source of early interleukin-4 following infection with third stage larvae of the filarial nematode Brugia pahangi. AB - Infection of C57BL/6 mice with the third-stage larvae of Brugia pahangi results in a rapid expansion of NK1.1(+) T cells in the spleen and draining lymph nodes. NK T cells produced interleukin-4 in the spleen within 24 h of infection, and these cells were CD4(-). PMID- 11895991 TI - Extended repertoire of genes encoding variable surface lipoproteins in Mycoplasma bovis strains. AB - A genomic cluster of vsp genes was previously shown to mediate high-frequency phenotypic switching of surface lipoprotein antigens in the bovine pathogen Mycoplasma bovis. This study revealed that field strains of M. bovis possess modified versions of the vsp gene complex in which extensive sequence variations occur primarily in the reiterated coding sequences of the vsp structural genes. These findings demonstrate that there is a vastly expanded potential for antigenic variation within populations of this organism. PMID- 11895992 TI - Human secretory immune response to fatty acid-binding protein fraction from Giardia lamblia. AB - The secretory immune response in humans infected with Giardia lamblia was studied by using saliva samples and an 8-kDa antigen capable of binding fatty acids. This antigen was not recognized by saliva samples from healthy individuals. The antigen may be useful in diagnostic studies of G. lamblia infection. PMID- 11895993 TI - Membrane localization contributes to the in vivo ADP-ribosylation of Ras by Pseudomonas aeruginosa ExoS. AB - Type III-delivered exoenzyme S (ExoS) preferentially ADP-ribosylated membrane associated His(6)HRas, relative to its cytosolic derivative His(6)HRas Delta CAAX. This indicates that the subcellular protein distribution contributes to in vivo ADP-ribosylation by ExoS. PMID- 11895994 TI - Complementation of a Treponema denticola flgE mutant with a novel coumermycin A1 resistant T. denticola shuttle vector system. AB - By using the mutated gyrB gene from a spontaneous coumermycin A1-resistant Treponema denticola, an Escherichia coli-T. denticola shuttle vector that renders T. denticola resistant to coumermycin was constructed. The complete T. denticola flgE gene was cloned into the shuttle vector pKMCou, and the vector was transformed into the T. denticola ATCC 33520 flgE erythromycin-resistant knockout mutant HL210. The coumermycin-resistant transformants were motile and restored FlgE activity. This complementation system should prove useful in studying the virulence factors of T. denticola and uncultivatible pathogenic spirochetes. PMID- 11895995 TI - Gel shift assay of nuclear extracts from Histoplasma capsulatum demonstrates the presence of several DNA binding proteins. AB - A gel shift assay was optimized to detect several general DNA binding proteins from Histoplasma capsulatum strain G217B. The electrophoretic mobility shift assay (EMSA) technique also detected protein(s) recognizing a pyrimidine-rich motif found in several Histoplasma promoters. Establishment of EMSA conditions provides an important framework to evaluate regulation of homeostatic or phase specific genes that may influence virulence in Histoplasma and other dimorphic fungal pathogens. PMID- 11895996 TI - Campylobacter protein glycosylation affects host cell interactions. AB - Campylobacter jejuni 81-176 pgl mutants impaired in general protein glycosylation showed reduced ability to adhere to and invade INT407 cells and to colonize intestinal tracts of mice. PMID- 11895997 TI - Role of Neisseria meningitidis luxS in cell-to-cell signaling and bacteremic infection. AB - Numerous pathogenic bacteria contain luxS, which is required for autoinducer-2 production. Here, we demonstrate that Neisseria meningitidis contains a functional copy of luxS that is necessary for full meningococcal virulence; strains with a luxS deletion are defective for bacteremia, a prerequisite of meningococcal pathogenesis. PMID- 11895998 TI - Molecular biology of thermoregulation. PMID- 11895999 TI - Hindlimb unloading rodent model: technical aspects. AB - Since its inception at the National Aeronautics and Space Administration (NASA) Ames Research Center in the mid-1970s, many laboratories around the world have used the rat hindlimb unloading model to simulate weightlessness and to study various aspects of musculoskeletal loading. In this model, the hindlimbs of rodents are elevated to produce a 30 degrees head-down tilt, which results in a cephalad fluid shift and avoids weightbearing by the hindquarters. Although several reviews have described scientific results obtained with this model, this is the first review to focus on the technical aspects of hindlimb unloading. This review includes a history of the technique, a brief comparison with spaceflight data, technical details, extension of the model to mice, and other important technical considerations (e.g., housing, room temperature, unloading angle, the potential need for multiple control groups, age, body weight, the use of the forelimb tissues as internal controls, and when to remove animals from experiments). This paper is intended as a reference for researchers, reviewers of manuscripts, and institutional animal care and use committees. Over 800 references, related to the hindlimb unloading model, can be accessed via the electronic version of this article. PMID- 11896000 TI - Early contributions of Russian stress and exercise physiologists. AB - In Russia, the free development of scientific ideas was suppressed in 1950 as a result of the actions of the Joint Session of the Academy of Sciences and the Academy of Medical Sciences of the USSR. Hans Selye's theory on the general adaptation syndrome was considered unscientific. From 1956 to 1958, Pjotr Anokhin and Pjotr Gorizontov presented definitive arguments for having the theory accepted by scientists while the significance of hormones in adaptation became a topic of endocrine studies (Boris Aleshin, Igor Eskin, Vassily Komissarenko, Samuel Leites, and Michael Kolpakov). Later, Felius Meerson made essential contributions to the adaptive significance of protein synthesis and stress limiting systems. The area of exercise physiology dealing with acute and chronic adaptation to strong physiological stressors was founded by Leon Orbeli and developed by Aleksei Krestovnikov. Significant contributors to this area were Vladimir Farfel, Nikolai Yakovlev, and Nikolai Zimkin. Although the majority of their publications have remained unknown outside of Russia, it is interesting that many of their results have been "rediscovered" by others. Yakovlev also deserves recognition because he was among the founders of contemporary exercise biochemistry and because his research has provided the foundation for current investigations. Several generations of young scientists have been inspired by the above-mentioned Russian scientists. Today, however, the research activities of scientists are no longer limited by political pressures but by financial resources instead. PMID- 11896001 TI - Activation of human plantar flexor muscles increases after electromyostimulation training. AB - Neuromuscular adaptations of the plantar flexor muscles were assessed before and subsequent to short-term electromyostimulation (EMS) training. Eight subjects underwent 16 sessions of isometric EMS training over 4 wk. Surface electromyographic (EMG) activity and torque obtained under maximal voluntary and electrically evoked contractions were analyzed to distinguish neural adaptations from contractile changes. After training, plantar flexor voluntary torque significantly increased under isometric conditions at the training angle (+8.1%, P < 0.05) and at the two eccentric velocities considered (+10.8 and +13.1%, P < 0.05). Torque gains were accompanied by higher normalized soleus EMG activity and, in the case of eccentric contractions, also by higher gastrocnemii EMG (P < 0.05). There was an 11.9% significant increase in both plantar flexor maximal voluntary activation (P < 0.01) and postactivation potentiation (P < 0.05), whereas contractile properties did not change after training. In the absence of a change in the control group, it was concluded that an increase in neural activation likely mediates the voluntary torque gains observed after short-term EMS training. PMID- 11896002 TI - Pulmonary gas exchange and acid-base state at 5,260 m in high-altitude Bolivians and acclimatized lowlanders. AB - Pulmonary gas exchange and acid-base state were compared in nine Danish lowlanders (L) acclimatized to 5,260 m for 9 wk and seven native Bolivian residents (N) of La Paz (altitude 3,600-4,100 m) brought acutely to this altitude. We evaluated normalcy of arterial pH and assessed pulmonary gas exchange and acid-base balance at rest and during peak exercise when breathing room air and 55% O2. Despite 9 wk at 5,260 m and considerable renal bicarbonate excretion (arterial plasma HCO3- concentration = 15.1 meq/l), resting arterial pH in L was 7.48 +/- 0.007 (significantly greater than 7.40). On the other hand, arterial pH in N was only 7.43 +/- 0.004 (despite arterial O2 saturation of 77%) after ascent from 3,600-4,100 to 5,260 m in 2 h. Maximal power output was similar in the two groups breathing air, whereas on 55% O2 only L showed a significant increase. During exercise in air, arterial PCO2 was 8 Torr lower in L than in N (P < 0.001), yet PO2 was the same such that, at maximal O2 uptake, alveolar arterial PO2 difference was lower in N (5.3 +/- 1.3 Torr) than in L (10.5 +/- 0.8 Torr), P = 0.004. Calculated O2 diffusing capacity was 40% higher in N than in L and, if referenced to maximal hyperoxic work, capacity was 73% greater in N. Buffering of lactic acid was greater in N, with 20% less increase in base deficit per millimole per liter rise in lactate. These data show in L persistent alkalosis even after 9 wk at 5,260 m. In N, the data show 1) insignificant reduction in exercise capacity when breathing air at 5,260 m compared with breathing 55% O2; 2) very little ventilatory response to acute hypoxemia (judged by arterial pH and arterial PCO2 responses to hyperoxia); 3) during exercise, greater pulmonary diffusing capacity than in L, allowing maintenance of arterial PO2 despite lower ventilation; and 4) better buffering of lactic acid. These results support and extend similar observations concerning adaptation in lung function in these and other high-altitude native groups previously performed at much lower altitudes. PMID- 11896003 TI - Gender influence on vasoactive hormones at rest and during a 70 degrees head-up tilt in healthy humans. AB - To evaluate the influence of age and gender on the neuroendocrine control of blood pressure in normal subjects, a 13-min 70 degrees head-up tilt (HUT) was applied after 3 h of recumbency to 109 healthy men and women aged 23-50 yr (age group I) and 51-77 yr (age group II). We found that age and gender had a significant influence on plasma norepinephrine (PNE) concentration at baseline and in the upright position. PNE was significantly higher in older men compared with the younger men and women of both age groups, suggesting a divergent age related activation of the sympathetic nervous system between genders at baseline as well as during a sustained orthostatic challenge. There was no significant influence of age or gender on plasma epinephrine at baseline or during HUT. Plasma renin activity was significantly higher at baseline as well as in the upright position during HUT in elderly men than in women. Age or gender had no influence on plasma vasopressin (PAVP), and, regardless of age, nonhypotensive HUT induced an extremely modest increase in PAVP. The syncopal subjects displayed a hormonal pattern associating increased PNE and a surge in plasma epinephrine and PAVP minutes before syncope during HUT. The orthostatic intolerance appears not to be a feature of healthy aging per se. In healthy subjects, both age and gender modulate markedly the cardiovascular and neuroendocrine responses to an orthostatic challenge and must be taken into consideration, particularly when catecholamine responses are studied. PMID- 11896004 TI - Exercise hyperpnea in chronic heart failure: relationships to lung stiffness and expiratory flow limitation. AB - The changes in breathing pattern and lung mechanics in response to incremental exercise were compared in 14 subjects with chronic heart failure and 15 normal subjects. In chronic heart failure subjects, exercise hyperpnea was achieved by increasing breathing frequency more than tidal volume. The rate of increase in breathing frequency with carbon dioxide output was inversely correlated (r = 0.61, P < 0.05) with dynamic lung compliance measured at rest, but not with static lung compliance either at rest or at maximum exercise. Although decrease in expiratory flow reserve near functional residual capacity in chronic heart failure occurred earlier with exercise than in the normal subjects (P < 0.01), it was not correlated with changes in breathing pattern or occurrence of tachypnea. Tachypnea was achieved in chronic heart failure subjects with an increase in duty cycle because of a greater than normal decrease in expiratory time with exercise. We conclude that in chronic heart failure preexisting increase in lung stiffness plays a significant role in causing tachypnea during exercise. The results of the present study do not support the hypothesis that dynamic compression of the airways downstream from the flow-limiting segment occurring during exercise contributes to hyperpnea. PMID- 11896006 TI - Effects of body position on the ventilatory response following an impulse exercise in humans. AB - The aim of this study was to identify some of the mechanisms that could be involved in blunted ventilatory response (VE) to exercise in the supine (S) position. The contribution of the recruitment of different muscle groups, the activity of the cardiac mechanoreceptors, the level of arterial baroreceptor stimulation, and the hemodynamic effects of gravity on the exercising muscles was analyzed during upright (U) and S exercise. Delayed rise in VE and pulmonary gas exchange following an impulselike change in work rate (supramaximal leg cycling at 240 W for 12 s) was measured in seven healthy subjects and six heart transplant patients both in U and S positions. This approach allows study of the relationship between the rise in VE and O2 uptake (VO2) without the confounding effects of contractions of different muscle groups. These responses were compared with those triggered by an impulselike change in work rate produced by the arms, which were positioned at the same level as the heart in S and U positions to separate effects of gravity on postexercising muscles from those on the rest of the body. Despite superimposable VO2 and CO2 output responses, the delayed VE response after leg exercise was significantly lower in the S posture than in the U position for each control subject and cardiac-transplant patient (-2.58 +/- 0.44 l and -3.52 +/- 1.11 l/min, respectively). In contrast, when impulse exercise was performed with the arms, reduction of ventilatory response in the S posture reached, at best, one-third of the deficit after leg exercise and was always associated with a reduction in VO2 of a similar magnitude. We concluded that reduction in VE response to exercise in the S position is independent of the types (groups) of muscles recruited and is not critically dependent on afferent signals originating from the heart but seems to rely on some of the effects of gravity on postexercising muscles. PMID- 11896005 TI - Double-stranded RNA causes airway hyperreactivity and neuronal M2 muscarinic receptor dysfunction. AB - Viral infection causes dysfunction of inhibitory M2 muscarinic receptors (M2Rs) on parasympathetic nerves, leading to airway hyperreactivity. The mechanisms of M2R dysfunction are incompletely understood. Double-stranded RNA (dsRNA), a product of viral replication, promotes the expression of interferons. Interferon gamma decreases M2R gene expression in cultured airway parasympathetic neurons. In this study, guinea pigs were treated with dsRNA (1 mg/kg ip) on 2 consecutive days. Twenty-four hours later, anesthetized guinea pigs had dysfunctional M2Rs and were hyperresponsive to electrical stimulation of the vagus nerves, in the absence of inflammation. DsRNA did not affect either cholinesterase or the function of postjunctional M3 muscarinic receptors on smooth muscle. M2Rs on the nerves supplying the heart were also dysfunctional, but M2Rs on the heart muscle itself functioned normally. Thus dsRNA causes increased bronchoconstriction and bradycardia via increased release of ACh from the vagus nerves because of loss of M2R function on parasympathetic nerves in the lungs and heart. Production of dsRNA may be a mechanism by which viruses cause dysfunction of neuronal M2Rs and airway hyperreactivity. PMID- 11896007 TI - Sympathetic activity and the heterogenous blood pressure response to exercise training in hypertensives. AB - To test whether changes in sympathetic nervous system (SNS) activity or insulin sensitivity contribute to the heterogeneous blood pressure response to aerobic exercise training, we used compartmental analysis of [3H]norepinephrine kinetics to determine the extravascular norepinephrine release rate (NE2) as an index of systemic SNS activity and determined the insulin sensitivity index (S(I)) by an intravenous glucose tolerance test, before and after 6 mo of aerobic exercise training, in 30 (63 +/- 7 yr) hypertensive subjects. Maximal O2 consumption increased from 18.4 +/- 0.7 to 20.8 +/- 0.7 ml x kg(-1) x min(-1) (P = 0.02). The average mean arterial blood pressure (MABP) did not change (114 +/- 2 vs. 114 +/- 2 mmHg); however, there was a wide range of responses (-19 to +17 mmHg). The average NE2 did not change significantly (2.11 +/- 0.15 vs. 1.99 +/- 0.13 microg x min(-1) x m(-2)), but there was a significant positive linear relationship between the change in NE2 and the change in MABP (r = 0.38, P = 0.04). S(I) increased from 2.81 +/- 0.37 to 3.71 +/- 0.42 microU x 10(-4) x min(-1) x ml(-1) (P = 0.004). The relationship between the change in S(I) and the change in MABP was not statistically significant (r = -0.03, P = 0.89). When the changes in maximal O2 consumption, percent body fat, NE2, and S(I) were considered as predictors of the change in MABP, only NE2 was a significant independent predictor. Thus suppression of SNS activity may play a role in the reduction in MABP and account for a portion of the heterogeneity of the MABP response to aerobic exercise training in older hypertensive subjects. PMID- 11896008 TI - Effect of the cytoskeletal prestress on the mechanical impedance of cultured airway smooth muscle cells. AB - We investigated the effect of the cytoskeletal prestress (P) on the elastic and frictional properties of cultured human airway smooth muscle cells during oscillatory loading; P is preexisting tensile stress in the actin cytoskeleton generated by the cell contractile apparatus. We oscillated (0.1 Hz, 6 Pa peak to peak) small ferromagnetic beads bound to integrin receptors and computed the storage (elastic) modulus (G') and the loss (frictional) modulus (G") from the applied torque and the corresponding bead rotation. All measurements were done at baseline and after cells were treated with graded doses of either histamine (0.1, 1, 10 microM) or isoproterenol (0.01, 0.1, 1, 10 microM). Values for P for these concentrations were taken from a previous study (Wang et al., Am J Physiol Cell Physiol, in press). It was found that G' and G", as well as P, increased/decreased with increasing doses of histamine/isoproterenol. Both G' and G" exhibited linear dependences on P: G'(Pa) = 0.20P + 82 and G"(Pa) = 0.05P + 32. The dependence of G' on P is consistent with our previous findings and with the behavior of stress-supported structures. The dependence of G" on P is a novel finding. It could be attributed to a variety of mechanisms. Some of those mechanisms are discussed in detail. We concluded that, in addition to the central mechanisms by which stress-supported structures develop mechanical stresses, other mechanisms might need to be invoked to fully explain the observed dependence of the cell mechanical properties on the state of cell contractility. PMID- 11896009 TI - Capillary supply of the tibialis anterior muscle in young, healthy, and moderately active men and women. AB - Tibialis anterior muscle biopsies from moderately active men and women (21-30 yr; n = 30) were examined to determine potential gender differences in capillarization. The fiber type proportions [type I (T1) approximately 73%] were unaffected by gender. The men (M) had significantly (P < 0.001) larger fibers than the women (W), with a greater gender effect for type II (T2) fibers (P < 0.001). The M and W had similar capillary densities (CD approximately 390 capillaries/mm2), but the capillaries-to-fiber ratio (C/F) was higher in the M (M = 2.20 +/- 0.35, W = 1.66 +/- 0.32; P < 0.01). Capillary contacts (CC) were higher in T2 than T1 for the M (P < 0.01), but not W, and M had greater CC (P < 0.001). Both fiber area per capillary (FA/C) and fiber perimeter per capillary (FP/C) indicated that T1 fibers had greater capillarization than T2 fibers (P < 0.001). There were no gender differences in T1 FA/C and T2 FA/C or T1 FP/C, but a gender difference existed for T2 FP/C (M = 60.5 +/- 10.9, W = 70.6 +/- 13.4; P < 0.01). The gender difference for C/F could be explained by fiber size; however, the physiological implications of the difference in T2 FP/C remains to be determined. In conclusion, despite gender differences for fiber size, overall, capillarization was similar between the men and women. PMID- 11896010 TI - Regular aerobic exercise and the age-related increase in carotid artery intima media thickness in healthy men. AB - Carotid artery intima-media thickness (IMT), an independent risk factor for stroke, increases with age. Habitual exercise is associated with a lower prevalence of stroke, but it is unclear whether this protective effect could be mediated through a favorable influence on carotid IMT. We examined this possibility using both cross-sectional and intervention approaches. First, 137 healthy men (age 18-77 yr) who were either sedentary or endurance trained were studied. In both groups, carotid IMT and IMT-to-lumen ratio were progressively higher with age (P < 0.05). There were no significant differences in measures of carotid IMT between sedentary and endurance-trained men at any age. Carotid systolic blood pressure increased progressively with age and was related to carotid IMT (r = 0.63, P < 0.01). Second, 18 healthy sedentary subjects (54 +/- 2 yr) were studied before and after 3 mo of endurance training. Carotid IMT, IMT/lumen ratio, and carotid systolic blood pressure did not change with exercise intervention. Our results do not support the hypothesis that regular aerobic exercise exerts its protective effect against stroke by attenuating the age related increase in carotid IMT. This lack of effect on carotid IMT may be due to the apparent inability of habitual exercise to prevent or reduce the age associated elevation in carotid distending pressure. PMID- 11896012 TI - Exercise training alters an anoxia-induced, glibenclamide-sensitive current in rat ventricular cardiocytes. AB - The effect of training on properties of a sarcolemmal ATP-sensitive K+ current (I(K(ATP))) was examined in left ventricular cardiocytes isolated from sedentary (Sed) and trained (Tr) female Sprague-Dawley rats. Whole cell patch-clamp techniques were used to characterize I(K(ATP)), an anoxia-inducible, glibencamide sensitive current. An anoxic condition was induced by superfusing cells with a buffer that was equilibrated with 100% N(2), maintained under a layer of argon, and that contained 2-deoxy-D-glucose. Over a 1-h period of anoxia, 59% of Tr cells and 85% Sed cells expressed I(K(ATP)). In those cells that did express I(K(ATP)), the time to expression of the current during the anoxic period occurred significantly later in cells from the Tr group compared with the Sed. Peak I(K(ATP)) density was significantly lower in the Tr cells compared with the Sed cells. These results indicate that the onset and magnitude of I(K(ATP)) were altered by training. These alterations in I(K(ATP)) may be reflective of processes that contribute to training-induced cardioprotection against ischemia reperfusion damage. PMID- 11896011 TI - Passive tension of rat skeletal soleus muscle fibers: effects of unloading conditions. AB - In this work we studied changes in passive elastic properties of rat soleus muscle fibers subjected to 14 days of hindlimb unloading (HU). For this purpose, we investigated the titin isoform expression in soleus muscles, passive tension fiber strain relationships of single fibers, and the effects of the thick filament depolymerization on passive tension development. The myosin heavy chain composition was also measured for all fibers studied. Despite a slow-to-fast transformation of the soleus muscles on the basis of their myosin heavy chain content, no modification in the titin isoform expression was detected after 14 days of HU. However, the passive tension-fiber strain relationships revealed that passive tension of both slow and fast HU soleus fibers increased less steeply with sarcomere length than that of control fibers. Gel analysis suggested that this result could be explained by a decrease in the amount of titin in soleus muscle after HU. Furthermore, the thick filament depolymerization was found to similarly decrease passive tension in control and HU soleus fibers. Taken together, these results suggested that HU did not change titin isoform expression in the soleus muscle, but rather modified muscle stiffness by decreasing the amount of titin. PMID- 11896014 TI - Neuromuscular fatigue during a long-duration cycling exercise. AB - The effects of prolonged cycling on neuromuscular parameters were studied in nine endurance-trained subjects during a 5-h exercise sustained at 55% of the maximal aerobic power. Torque during maximal voluntary contraction (MVC) of the quadriceps muscle decreased progressively throughout the exercise (P < 0.01) and was 18% less at the end of exercise compared with the preexercise value. Peak twitch torque, contraction time, and total area of mechanical response decreased significantly (P < 0.05) after the first hour of exercise. In contrast, changes in M-wave characteristics were significant only after the fourth hour of the exercise. Significant reductions (P < 0.05) in electromyographic activity normalized to the M wave occurred after the first hour for the vastus lateralis muscle but only at the end of the exercise for the vastus medialis muscle. Muscle activation level, assessed by the twitch interpolation technique, decreased by 8% (P < 0.05) at the end of the exercise. The results suggest that the time course is such that the contractile properties are significantly altered after the first hour, whereas excitability and central drive are more impaired toward the latter stages of the 5-h cycling exercise. PMID- 11896013 TI - Chronic hypoxia upregulates connexin43 expression in rat carotid body and petrosal ganglion. AB - Recent studies have demonstrated that oxygen-sensitive type I cells in the carotid body express the gap junction-forming protein connexin43 (Cx43). In the present study, we examined the hypothesis that chronic exposure to hypoxia increases Cx43 expression in type I cells as well as in chemoafferent neurons in the petrosal ganglion. Immunocytochemical studies in tissues from normal rats revealed diffuse and granular Cx43-like immunoreactivity in the cytoplasm of type I cells and dense punctate spots of immunoreactive product at the margins of type I cells and near the borders of chemosensory cell lobules. Cx43-like immunoreactivity was not detectable in petrosal ganglion neurons from normal animals. After a 2-wk exposure to hypobaric (380 Torr) hypoxia, Cx43 immunostaining was substantially enhanced in and around type I cells. Moreover, chronic hypoxia elicited the expression of Cx43-like immunoreactivity in the cytoplasm of afferent neurons throughout the petrosal ganglion. Quantitative RT PCR studies indicate that chronic hypoxia evokes a substantial increase in Cx43 mRNA levels in the carotid body, along with a marked elevation of Cx43 expression in the petrosal ganglion. Increased Cx43 expression and gap junction formation in type I cells and sensory neurons may contribute to carotid body adaptation during sustained stimulation in extreme physiological conditions. PMID- 11896015 TI - Effect of pregnancy on joint contracture in the rat knee. AB - As there is evidence that ligamentous laxity is affected by the female hormones, we hypothesized that hormonal changes occurring during pregnancy could have a therapeutic role in preventing the development of a joint contracture. Knee joint contractures were created in pregnant and nonpregnant rats. After 2 wk of immobilization, the degree of contracture was measured with structural properties of the medial collateral and anterior cruciate ligaments and the pubic symphysis. Although not statistically significant, there was a general trend toward reduced contracture in pregnant compared with nonpregnant rats. Cutting the posterior capsule significantly decreased contracture for both the pregnant and nonpregnant groups, confirming the contribution of capsular structures to contracture. Ultimate loads of the medial collateral and anterior cruciate ligaments significantly decreased after immobilization compared with control, but there was no significant effect due to pregnancy. Stiffness and ultimate load of the pubic symphysis were not significantly different between pregnant and nonpregnant groups. The trend toward reduced contracture with pregnancy points toward a possible therapeutic role for female hormones in the prevention of postoperative and/or posttraumatic joint contracture. PMID- 11896016 TI - Mechanistic basis for the gas exchange threshold in Thoroughbred horses. AB - The exercising Thoroughbred horse (TB) is capable of exceptional cardiopulmonary performance. However, because the ventilatory equivalent for O2 (VE/VO2) does not increase above the gas exchange threshold (Tge), hypercapnia and hypoxemia accompany intense exercise in the TB compared with humans, in whom VE/VO2 increases during supra-Tge work, which both removes the CO2 produced by the HCO buffering of lactic acid and prevents arterial partial pressure of CO2 (PaCO2) from rising. We used breath-by-breath techniques to analyze the relationship between CO2 output (VCO2) and VO2 [V-slope lactate threshold (LT) estimation] during an incremental test to fatigue (7 to approximately 15 m/s; 1 m x s(-1) x min(-1)) in six TB. Peak blood lactate increased to 29.2 +/- 1.9 mM/l. However, as neither VE/VO2 nor VE/VCO2 increased, PaCO2 increased to 56.6 +/- 2.3 Torr at peak VO2 (VO2 max). Despite the presence of a relative hypoventilation (i.e., no increase in VE/VO2 or VE/VCO2), a distinct Tge was evidenced at 62.6 +/- 2.7% VO2 max. Tge occurred at a significantly higher (P < 0.05) percentage of VO2 max than the lactate (45.1 +/- 5.0%) or pH (47.4 +/- 6.6%) but not the bicarbonate (65.3 +/- 6.6%) threshold. In addition, PaCO2 was elevated significantly only at a workload > Tge. Thus, in marked contrast to healthy humans, pronounced V-slope (increase VCO2/VO2) behavior occurs in TB concomitant with elevated PaCO2 and without evidence of a ventilatory threshold. PMID- 11896017 TI - Effects of hypothyroidism on maximum specific force in rat diaphragm muscle fibers. AB - We hypothesized that 1) hypothyroidism (Hyp) decreases myosin heavy chain (MHC) content per half-sarcomere in diaphragm muscle (Dia(m)) fibers, 2) Hyp decreases the maximum specific force (F(max)) of Dia(m) fibers because of the reduction in MHC content per half-sarcomere, and 3) Hyp affects MHC content per half-sarcomere and F(max) to a greater extent in fibers expressing MHC type 2X (MHC(2X)) and/or MHC type 2B (MHC(2B)). Studies were performed on single Triton X-permeabilized fibers activated at pCa 4.0. MHC content per half-sarcomere was determined by densitometric analysis of SDS-polyacrylamide gels and comparison with a standard curve of known MHC concentrations. After 3 wk of Hyp, MHC content per half sarcomere was reduced in fibers expressing MHC(2X) and/or MHC(2B). On the basis of electron-microscopic analysis, this reduction in MHC content was also reflected by a decrease in myofibrillar volume density and thick filament density. Hyp decreased F(max) across all MHC isoforms; however, the greatest decrease occurred in fibers expressing fast MHC isoforms (approximately 40 vs. approximately 20% for fibers expressing slow MHC isoforms). When normalized for MHC content per half-sarcomere, force generated by Hyp fibers expressing MHC(2A) was reduced compared with control fibers, whereas force per half-sarcomere MHC content was higher for fibers expressing MHC(2X) and/or MHC(2B) in the Hyp Dia(m) than for controls. These results indicate that the effect of Hyp is more pronounced on fibers expressing MHC(2X) and/or MHC(2B) and that the reduction of F(max) with Hyp may be at least partially attributed to a decrease in MHC content per half-sarcomere but not to changes in force per cross bridge. PMID- 11896018 TI - H1-receptor antagonist, tripelennamine, does not affect arterial hypoxemia in exercising Thoroughbreds. AB - It has been suggested that pulmonary injury and inflammation-induced histamine release from airway mast cells may contribute to exercise-induced arterial hypoxemia (EIAH). Because stress failure of pulmonary capillaries and EIAH are routinely observed in exercising horses, we examined whether preexercise administration of an H1-receptor antagonist may mitigate EIAH. Two sets of experiments, placebo (saline) and antihistaminic (tripelennamine HCl at 1.10 mg/kg iv, 15 min preexercise) studies, were carried out on seven healthy, exercise-trained Thoroughbred horses in random order 7 days apart. Arterial and mixed venous blood-gas and pH measurements were made at rest before and after saline or drug administration and during incremental exercise leading to maximal exertion at 14 m/s on 3.5% uphill grade for 120 s. Galloping at this workload elicited maximal heart rate and induced exercise-induced pulmonary hemorrhage in all horses in both treatments, thereby indicating that capillary stress failure related pulmonary injury had occurred. In both treatments, EIAH, desaturation of hemoglobin, hypercapnia, and acidosis of a similar magnitude developed during maximal exertion, and statistically significant differences between the placebo and antihistaminic studies could not be demonstrated. The failure of the H1 receptor antagonist to modify EIAH significantly suggests that pulmonary injury induced histamine release may not play a major role in bringing about EIAH in Thoroughbred horses. PMID- 11896019 TI - Exercise training normalizes altered calcium-handling proteins during development of heart failure. AB - The cardiac sarcoplasmic reticulum calcium-ATPase (SERCA2a), Na+/Ca2+ exchanger (NCX1), and ryanodine receptor (RyR2) are proteins involved in the regulation of myocyte calcium. We tested whether exercise training (ET) alters those proteins during development of chronic heart failure (CHF). Ten dogs were chronically instrumented to permit hemodynamic measurements. Five dogs underwent 4 wk of cardiac pacing (210 beats/min for 3 wk and 240 beats/min for the 4th wk), whereas five dogs underwent the same pacing regimen plus daily ET (5.1 +/- 0.3 km/h, 2 h/day). Paced animals developed CHF characterized by hemodynamic abnormalities and reduced ejection fraction. ET preserved resting hemodynamics and ejection fraction. Left ventricular samples were obtained from all dogs and another five normal dogs for mRNA (Northern analysis, band intensities normalized to glyceraldehyde-3-phosphate dehydrogenase) and protein level (Western analysis, band intensities normalized to tubulin) measurements. In failing hearts, SERCA2a was decreased by 33% (P < 0.05) and 65% (P < 0.05) in mRNA and protein level, respectively, compared with normal hearts; there was only an 8.6% reduction in mRNA and a 32% reduction in protein in exercised animals (P < 0.05 from CHF). mRNA expression of NCX1 increased by 44% in paced-only dogs compared with normal (P < 0.05) but only by 22% in trained dogs (P < 0.05 vs. CHF); protein level of NCX1 was elevated in paced-only dogs (71%, P < 0.05) but partially normalized by ET (33%, P < 0.05 from CHF). RyR2 was not altered in any of the dogs. In conclusion, long-term ET may ameliorate cardiac deterioration during development of CHF, in part via normalization of myocardial calcium-handling proteins. PMID- 11896020 TI - Estimation of lung liquid production in fetal sheep with blue dye dextran and radioiodinated serum albumin. AB - Lung liquid production and reabsorption rates and lung volumes were measured in 99 fetal sheep (119-148 days of gestation) by indicator-dilution methods with the simultaneous use of blue dye dextran (BDD) and radioiodinated serum albumin (RISA). There were no significant differences between rates of lung liquid production or reabsorption by the two methods (n = 71 pairs; paired t-test; Wilcoxon test; ANOVA); this was equally true for rates in milliliters per hour or milliliters per kilogram body weight per hour and was independent of age. Volumes measured by both methods showed a close linear relationship (r = 0.97; for slope P < 0.0001; n = 99), whether expressed as milliliters or milliliters per kilogram body weight. Either method could give the higher volume. Values differed by only approximately 4%, independent of age or parameter (ml or ml/kg body wt; volumes regressed to original volume, or as measured in untreated control hours). However, this small difference was significant by paired t-test or Wilcoxon test when all data were combined irrespective of age; it was not significant after allowance for gestational age (two-way ANOVA). Both indicators showed the same increase in lung volume toward birth and the same fall when related to body weight (slopes significant P = 0.0003-0.0004; r = 0.93). Two-way ANOVA showed that the declines were significant (P = 0.003). The data suggest that 1) there was no significant difference in production or reabsorption rates measured by BDD or RISA, 2) differences in volumes measured by the two indicators were only significant if gestational age was ignored and were too small to have physiological importance, and 3) although BDD and RISA each may have methodological weaknesses, for purposes of measuring lung liquid volumes both are sufficiently accurate and reproducible to obtain meaningful physiological results. PMID- 11896021 TI - Effects of expiratory muscle work on muscle sympathetic nerve activity. AB - We hypothesized that contractions of the expiratory muscles carried out to the point of task failure would cause an increase in muscle sympathetic nerve activity (MSNA). We measured MSNA directly in six healthy men during resisted expiration (60% maximal expiratory pressure) leading to task failure with long [breathing frequency (f(b)) = 15 breaths/min; expiratory time (TE)/total respiratory cycle duration (TT) = 0.7] and short (f(b) = 30 breaths/min; TE/TT = 0.4) TE. Both of these types of expiratory muscle contractions elicited time dependent increases in MSNA burst frequency that averaged +139 and +239%, respectively, above baseline at end exercise. The increased MSNA coincided with increases in mean arterial pressure (MAP) for both the long-TE (+28 +/- 6 mmHg) and short-TE (+22 +/- 14 mmHg) trials. Neither MSNA nor MAP changed when the breathing patterns and increased tidal volume of the task failure trials were mimicked without resistance or task failure. Furthermore, very high levels of expiratory motor output (95% maximal expiratory pressure; f(b) = 12 breaths/min; TE/TT = 0.35) and high rates of expiratory flow and expiratory muscle shortening without task failure (no resistance; f(b) = 45 breaths/min; TE/TT = 0.4; tidal volume = 1.9 x eupnea) had no effect on MSNA or MAP. Within-breath analysis of the short-expiration trials showed augmented MSNA at the onset of and throughout expiration that was consistent with an influence of high levels of central expiratory motor output. Thus high-intensity contractions of expiratory muscles to the point of task failure caused a time-dependent sympathoexcitation; these effects on MSNA were similar in their time dependency to those caused by high intensity rhythmic contractions of the diaphragm and forearm muscles taken to the point of task failure. The evidence suggests that these effects are mediated primarily via a muscle metaboreflex with a minor, variable contribution from augmented central expiratory motor output. PMID- 11896022 TI - Plasma corticosterone response to acute and chronic voluntary exercise in female house mice. AB - Plasma levels of corticosterone (B) respond acutely to exercise in all mammals that have been studied, but the literature contains conflicting reports regarding how chronic activity alters this response. We measured acute and chronic effects of voluntary activity on B in a novel animal model, mice selectively bred for high voluntary wheel running. Female mice were housed with or without wheels for 8 wk beginning at 26 days of age. Wheel-access selection mice had significantly higher B at night 8, day 15, and night 29, compared with wheel-access controls. Elevation of B was an acute effect of voluntary exercise. When adjusted for running in the previous 20 min, no difference between wheel-access selection and control animals remained. No training effect on B response was observed. These results are among the strongest evidence that, in some animals, the acute B response is unaffected by chronic voluntary exercise. In mice without wheels, selection mice had significantly higher B than controls at day 15, night 29, and night 50, suggesting that selection resulted in a modulation of the hypothalamic pituitary-adrenal axis. Growth over the first 4 wk of treatment was significantly and inversely related to average night B levels within each of the four treatment groups. PMID- 11896023 TI - Heat stress increases muscle glycogen use but reduces the oxidation of ingested carbohydrates during exercise. AB - The aim of the present study was to test the hypothesis that the oxidation rate of ingested carbohydrate (CHO) is impaired during exercise in the heat compared with a cool environment. Nine trained cyclists (maximal oxygen consumption 65 +/- 1 ml x kg body wt(-1) x min(-1)) exercised on two different occasions for 90 min at 55% maximum power ouptput at an ambient temperature of either 16.4 +/- 0.2 degrees C (cool trial) or 35.4 +/- 0.1 degrees C (heat trial). Subjects received 8% glucose solutions that were enriched with [U-13C]glucose for measurements of exogenous glucose, plasma glucose, liver-derived glucose and muscle glycogen oxidation. Exogenous glucose oxidation during the final 30 min of exercise was significantly (P < 0.05) lower in the heat compared with the cool trial (0.76 +/- 0.06 vs. 0.84 +/- 0.05 g/min). Muscle glycogen oxidation during the final 30 min of exercise was increased by 25% in the heat (2.07 +/- 0.16 vs. 1.66 +/- 0.09 g/min; P < 0.05), and liver-derived glucose oxidation was not different. There was a trend toward a higher total CHO oxidation and a lower plasma glucose oxidation in the heat although this did not reach statistical significance (P = 0.087 and P = 0.082, respectively). These results demonstrate that the oxidation rate of ingested CHO is reduced and muscle glycogen utilization is increased during exercise in the heat compared with a cool environment. PMID- 11896024 TI - Changes in MCT 1, MCT 4, and LDH expression are tissue specific in rats after long-term hypobaric hypoxia. AB - Little is known about the effect of chronic hypobaric hypoxia on the enzymes and transporters involved in lactate metabolism. We looked at the protein expression of monocarboxylate transporters MCT 1, MCT 2, and MCT 4, along with total lactate dehydrogenase (LDH) and LDH isozymes in skeletal muscle, cardiac muscle, and liver. Expression of these components of the lactate shuttle affects the ability to transport and oxidize lactate. We hypothesized that the expression of MCTs and LDH would increase after acclimation to high altitude (HA). The response to acclimation to HA was, however, tissue specific. In addition, the response was different in whole muscle (Mu) and mitochondria-enriched (Mi) fractions. Heart, soleus, and plantaris muscles showed the greatest response to HA. Acclimation resulted in a 34% increase in MCT 4 in heart and a decrease in MCT 1 (-47%) and MCT 4 (-47%) in plantaris Mu. In Mi fractions, the heart had an increase (+40%) and soleus a decrease (-40%) in LDH. HA also had a significant effect on the LDH isozyme composition of both the Mu and Mi fractions. Mitochondrial density was decreased in both the soleus (-17%) and plantaris (-44%) as a result of chronic hypoxia. We conclude that chronic hypoxia had a tissue-specific effect on MCTs and LDH (that form the lactate shuttle) but did not produce a consistent increase in these components in all tissues. PMID- 11896025 TI - Human neuromuscular fatigue is associated with altered Na+-K+-ATPase activity following isometric exercise. AB - The purpose of this study was to investigate the hypothesis that reductions in Na+-K+- ATPase activity are associated with neuromuscular fatigue following isometric exercise. In control (Con) and exercised (Ex) legs, force and electromyogram were measured in 14 volunteers [age, 23.4 +/- 0.7 (SE) yr] before and immediately after (PST0), 1 h after (PST1), and 4 h after (PST4) isometric, single-leg extension exercise at ~60% of maximal voluntary contraction for 30 min using a 0.5 duty cycle (5-s contraction, 5-s rest). Tissue was obtained from vastus lateralis muscle before exercise in Con and after exercise in both the Con (PST0) and Ex legs (PST0, PST1, PST4), for the measurements of Na+-K+-ATPase activity, as determined by the 3-O-methylfluorescein phosphatase (3-O-MFPase) assay. Voluntary (maximal voluntary contraction) and elicited (10, 20, 50, 100 Hz) force was reduced 30-55% (P < 0.05) at PST0 and did not recover by PST4. Muscle action potential (M-wave) amplitude and area (measured in the vastus medialis) and 3-O-MFPase activity at PST0-Ex were less than that at PST0-Con (P < 0.05) by 37, 25, and 38%, respectively. M-wave area at PST1-Ex was also less than that at PST1-Con (P < 0.05). Changes in 3-O-MFPase activity correlated to changes in M-wave area across all time points (r = 0.38, P < 0.05, n = 45). These results demonstrate that Na+-K+- ATPase activity is reduced by sustained isometric exercise in humans from that in a matched Con leg and that this reduction in Na+ K+-ATPase activity is associated with loss of excitability as indicated by M-wave alterations. PMID- 11896026 TI - Role of capacitative Ca2+ entry in bronchial contraction and remodeling. AB - Asthma is characterized by airway inflammation, bronchial hyperresponsiveness, and airway obstruction by bronchospasm and bronchial wall thickening due to smooth muscle hypertrophy. A rise in cytosolic free Ca2+ concentration ([Ca2+]cyt) may serve as a shared signal transduction element that causes bronchial constriction and bronchial wall thickening in asthma. In this study, we examined whether capacitative Ca2+ entry (CCE) induced by depletion of intracellular Ca2+ stores was involved in agonist-mediated bronchial constriction and bronchial smooth muscle cell (BSMC) proliferation. In isolated bronchial rings, acetylcholine (ACh) induced a transient contraction in the absence of extracellular Ca2+ because of Ca2+ release from intracellular Ca2+ stores. Restoration of extracellular Ca2+ in the presence of atropine, an M-receptor blocker, induced a further contraction that was apparently caused by a rise in [Ca2+]cyt due to CCE. In single BSMC, amplitudes of the store depletion-activated currents (I(SOC)) and CCE were both enhanced when the cells proliferate, whereas chelation of extracellular Ca2+ with EGTA significantly inhibited the cell growth in the presence of serum. Furthermore, the mRNA expression of TRPC1, a transient receptor potential channel gene, was much greater in proliferating BSMC than in growth-arrested cells. Blockade of the store-operated Ca2+ channels by Ni2+ decreased I(SOC) and CCE and markedly attenuated BSMC proliferation. These results suggest that upregulated TRPC1 expression, increased I(SOC), enhanced CCE, and elevated [Ca2+]cyt may play important roles in mediating bronchial constriction and BSMC proliferation. PMID- 11896027 TI - Silver ions induce Ca2+ release from the SR in vitro by acting on the Ca2+ release channel and the Ca2+ pump. AB - Silver nitrate (AgNO3) is a sulfhydryl oxidizing agent that induces a biphasic Ca2+ release from isolated sarcoplasmic reticulum (SR) vesicles by presumably oxidizing critical sulfhydryl groups in the Ca2+ release channel (CRC), causing the channel to open. To further examine the effects of AgNO3 on the CRC and the Ca2+-ATPase, Ca2+ release was measured in muscle homogenates prepared from rat hindlimb muscle using indo 1. Cyclopiazonic acid (CPA) and ruthenium red (RR) were used to inhibit the Ca2+-ATPase and block the CRC, respectively, before inducing Ca2+ release with both AgNO3 and 4-chloro-m-cresol (4-CMC), a releasing agent specific for the CRC. With AgNO3 and CPA, the early rapid rate of release (phase 1) was increased (P < 0.05) by 42% (314 +/- 5 vs. 446 +/- 39 micromol x g protein(-1) x min(-1)), whereas the slower, more prolonged rate of release (phase 2) was decreased (P < 0.05) by 72% (267 +/- 39 vs. 74 +/- 7.7 micromol x g protein(-1) x min(-1)). RR, in combination with AgNO3, had no effect on phase 1 (P > 0.05) (314 +/- 51 vs. 334 +/- 43 micromol x g protein(-1) x min(-1)) and decreased phase 2 (P < 0.05) by 65% (245 +/- 34 vs. 105 +/- 8.2 micromol x g protein(-1) x min(-1)). With 4-CMC, CPA had no effect (P > 0.05) on either phase 1 or 2. With addition of RR, phase 1 was reduced (P < 0.05) by 59% (2,468 +/- 279 vs. 1,004 +/- 87 micromol x g protein(-1) x min(-1)), and RR completely blocked phase 2. Both AgNO3 and 4-CMC fully inhibited Ca2+-ATPase activity measured in homogenates. These findings indicate that AgNO3, but not 4-CMC, induces Ca2+ release by acting on both the CRC and the Ca2+-ATPase. PMID- 11896028 TI - Gender differences in regional body composition and somatotrophic influences of IGF-I and leptin. AB - This study evaluated the arm, trunk, and leg for fat mass, lean soft tissue mass, and bone mineral content (BMC) assessed via dual-energy X-ray absorptiometry in a group of age-matched (approximately 29 yr) men (n = 57) and women (n = 63) and determined their relationship to insulin-like growth factor I (IGF-I) and leptin. After analysis of covariance adjustment to control for differences in body mass between genders, the differences that persisted (P < or = 0.05) were for lean soft tissue mass of the arm (men: 7.1 kg vs. women: 6.4 kg) and fat mass of the leg (men: 5.3 kg vs. women: 6.8 kg). Men and women had similar (P > or = 0.05) values for fat mass of the arms and trunk and lean soft tissue mass of the legs and trunk. Serum IGF-I and insulin-like growth factor binding protein-3 correlated (P < or = 0.05) with all measures of BMC (r values ranged from 0.31 to 0.39) and some measures of lean soft tissue mass for women (r = 0.30) but not men. Leptin correlated (P < or = 0.05) similarly for measures of fat mass for both genders (r values ranging from 0.74 to 0.85) and for lean soft tissue mass of the trunk (r = 0.40) and total body (r = 0.32) for men and for the arms in women (r = 0.56). These data demonstrate that 1) the main phenotypic gender differences in body composition are that men have more of their muscle mass in their arms and women have more of their fat mass in their legs and 2) gender differences exist in the relationship between somatotrophic hormones and lean soft tissue mass. PMID- 11896029 TI - Effects of confinement (110 and 240 days) on neuroendocrine stress response and changes of immune cells in men. AB - The aim of the study was to evaluate the effects of long-term confinement on stress-permissive neuroendocrine and immune responses in humans. Two groups of four male subjects were confined 240 days (group 240) or 110 days (group 110) in two space modules of 100 or 200 m3, respectively. During confinement, none of the volunteers developed psychic stress as could be examined and verified by a current stress test. However, in group 240 but not in group 110, the diurnal rhythm of cortisol secretion was slightly depressed and the urine excretion of norepinephrine significantly increased. The innate part of the immune system became activated as seen by a rise in the number of circulating granulocytes and the enhanced expression of beta2-integrins. In contrast, the ratio of T-helper to T-suppressor cells decreased. All these effects, observed during confinement, were even more pronounced in both groups when values of endocrinological and immunological parameters were compared between before and 1 wk after the end of the confinement period. Hence, return to normal life exerts pronounced effects to a much higher degree, irrespective of how long or under which conditions individuals were confined. Because the delayed-type hypersensitivity skin reaction against recall antigens remained unaffected, it is to be presumed that confinement appears to induce distinct sympathoadrenergic activation and immunological changes but no clinically relevant immunosuppression. PMID- 11896030 TI - MLR stimulation and exercise pressor reflex activate different renal sympathetic fibers in decerebrate cats. AB - Although mesencephalic locomotor region (MLR) stimulation and the exercise pressor reflex have been shown to increase whole nerve renal sympathetic activity, it is not known whether these mechanisms converge onto the same population of renal sympathetic postganglionic efferents. In decerebrate cats, we examined the responses of single renal sympathetic postganglionic efferents to stimulation of the MLR and the exercise pressor reflex (i.e., static contraction of the triceps surae muscles). We found that, in most instances (24 of 28 fibers), either MLR stimulation or the muscle reflex, but not both, increased the discharge of renal postganglionic sympathetic efferents. In addition, we found that renal sympathetic efferents that responded to static contraction while the muscles were freely perfused responded more vigorously to static contraction during circulatory arrest. Moreover, stretch of the calcaneal (Achilles) tendon stimulated the same renal sympathetic efferents as did static contraction. These findings suggest that MLR stimulation and the exercise pressor reflex do not converge onto the same renal sympathetic postganglionic efferents. PMID- 11896031 TI - Estrogen attenuates the cardiovascular and ventilatory responses to central command in cats. AB - Static exercise is well known to increase heart rate, arterial blood pressure, and ventilation. These increases appear to be less in women than in men, a difference that has been attributed to an effect of estrogen on neuronal function. In decerebrate male cats, we examined the effect of estrogen (17beta estradiol; 0.001, 0.01, 0.1, and 1.0 microg/kg iv) on the cardiovascular and ventilatory responses to central command and the exercise pressor reflex, the two neural mechanisms responsible for evoking the autonomic and ventilatory responses to exercise. We found that 17beta-estradiol, in each of the three doses tested, attenuated the pressor, cardioaccelerator, and phrenic nerve responses to electrical stimulation of the mesencephalic locomotor region (i.e., central command). In contrast, none of the doses of 17beta-estradiol had any effect on the pressor, cardioaccelerator, and ventilatory responses to static contraction or stretch of the triceps surae muscles. We conclude that, in decerebrate male cats, estrogen injected intravenously attenuates cardiovascular and ventilatory responses to central command but has no effect on responses to the exercise pressor reflex. PMID- 11896032 TI - On the respiratory function of the ribs. AB - To assess the respiratory function of the ribs, we measured the changes in airway opening pressure (Pao) induced by stimulation of the parasternal and external intercostal muscles in anesthetized dogs, first before and then after the bony ribs were removed from both sides of the chest. Stimulating either set of muscles with the rib cage intact elicited a fall in Pao in all animals. After removal of the ribs, however, the fall in Pao produced by the parasternal intercostals was reduced by 60% and the fall produced by the external intercostals was eliminated. The normal outward curvature of the rib cage was also abolished in this condition, and when the curvature was restored by a small inflation, external intercostal stimulation consistently elicited a rise rather than a fall in Pao. These findings thus confirm that the ribs play a critical role in the act of breathing by converting intercostal muscle shortening into lung volume expansion. In addition, they carry the compression that is required to balance the pressure difference across the chest wall. PMID- 11896033 TI - Muscle pump-dependent self-perfusion mechanism in legs in normal subjects and patients with heart failure. AB - Leg venous pressure markedly falls during upright exercise via a muscle pump effect, creating de novo perfusion pressure. We examined physiological roles of this mechanism in increasing femoral artery blood flow (FABF) and its alterations in chronic heart failure (CHF). In 10 normal subjects and 10 patients with CHF, standard hemodynamic variables, mean ankle vein pressure (MAVP), and FABF with Doppler techniques were obtained during graded upright bicycle exercise. To evaluate a nonspecific blood flow response, normal subjects also performed supine exercise. In normal subjects, MAVP rapidly declined by 45 mmHg and FABF correspondingly increased 5.3-fold without a systemic pressor response during 10 s of light upright exercise at 5 W. Approximately 67% of the blood flow response was attributed to the venous pressure drop-dependent mechanism. In CHF patients, MAVP declined by only 36 mmHg and FABF increased only 1.7-fold during the same upright exercise. The muscle venous pump has an ability to increase FABF at least threefold via the venous pressure drop-dependent mechanism. This mechanism is impaired in CHF patients. PMID- 11896035 TI - Phosphorylation of the regulatory light chains of myosin affects Ca2+ sensitivity of skeletal muscle contraction. AB - The role of phosphorylation of the myosin regulatory light chains (RLC) is well established in smooth muscle contraction, but in striated (skeletal and cardiac) muscle its role is still controversial. We have studied the effects of RLC phosphorylation in reconstituted myosin and in skinned skeletal muscle fibers where Ca2+ sensitivity and the kinetics of steady-state force development were measured. Skeletal muscle myosin reconstituted with phosphorylated RLC produced a much higher Ca2+ sensitivity of thin filament-regulated ATPase activity than nonphosphorylated RLC (change in -log of the Ca2+ concentration producing half maximal activation = approximately 0.25). The same was true for the Ca2+ sensitivity of force in skinned skeletal muscle fibers, which increased on reconstitution of the fibers with the phosphorylated RLC. In addition, we have shown that the level of endogenous RLC phosphorylation is a crucial determinant of the Ca2+ sensitivity of force development. Studies of the effects of RLC phosphorylation on the kinetics of force activation with the caged Ca2+, DM nitrophen, showed a slight increase in the rates of force development with low statistical significance. However, an increase from 69 to 84% of the initial steady-state force was observed when nonphosphorylated RLC-reconstituted fibers were subsequently phosphorylated with exogenous myosin light chain kinase. In conclusion, our results suggest that, although Ca2+ binding to the troponin tropomyosin complex is the primary regulator of skeletal muscle contraction, RLC play an important modulatory role in this process. PMID- 11896034 TI - Physiology of sulfide in the rat colon: use of bismuth to assess colonic sulfide production. AB - Colonic bacteria produce hydrogen sulfide, a toxic compound postulated to play a pathogenetic role in ulcerative colitis. Colonic sulfide exposure has previously been assessed via measurements of fecal sulfide concentration. However, we found that <1% of fecal sulfide of rats was free, the remainder being bound in soluble and insoluble complexes. Thus fecal sulfide concentrations may reflect sulfide binding capacity rather than the toxic potential of feces. We utilized bismuth subnitrate to quantitate intracolonic sulfide release based on observations that bismuth 1) avidly binds sulfide; 2) quantitatively releases bound sulfide when acidified; and 3) does not influence fecal sulfide production by fecal homogenates. Rats ingesting bismuth subnitrate excreted 350 +/- 18 micromol/day of fecal sulfide compared with 9 +/- 1 micromol/day in control rats. Thus the colon normally absorbs approximately 340 micromol of sulfide daily, a quantity that would produce local and systemic injury if not efficiently detoxified by the colonic mucosa. Studies utilizing bismuth should help to clarify the factors influencing sulfide production in the human colon. PMID- 11896036 TI - Dynamic time course of hemodynamic responses after passive head-up tilt and tilt back to supine position. AB - Mechanisms involved in the control of arterial pressure during postural changes were studied by analysis of the dynamic time course of cardiovascular changes during head-up tilt (HUT) and tilt back to supine position (TB). Beat-to-beat values of cardiovascular variables were recorded continuously before, during, and after passive HUT to 30 degrees in seven healthy humans. Left cardiac stroke volume (SV, Doppler ultrasound), mean arterial blood pressure (MAP), heart rate (HR), cardiac output (CO), and total peripheral conductance (TPC) were recorded. During HUT, MAP at the level of the carotid baroreceptors decreased by approximately 5 mmHg. There was a striking asymmetry between the time courses of cardiovascular changes on HUT and on TB. Adjustments generally took up to 30 s after HUT, whereas most changes were completed during the first 10 s after TB. Cardiovascular reflex adjustments of HR and TPC were more symmetrical. After HUT, SV was maintained during the first 4-6 s and then decreased steadily during the next 30 s to a stable level approximately 25% below its pretilt value. However, after TB, SV increased rapidly to its pretilt value in <10 s. This asymmetry in SV dynamics may be explained in part by a more rapid change in left cardiac filling after TB than after HUT. On TB, there must be a rapid inflow of stagnant blood from the legs, whereas venous valves will impede backward filling of veins in the lower body on HUT. In conclusion, we have revealed a characteristic asymmetry in cardiovascular responses to inverse variations in gravity forces in humans. This asymmetry can be explained in part by nonlinear, hydrodynamic factors, such as the one-way effect of venous valves in the lower part of the body. PMID- 11896037 TI - Attenuated hepatosplanchnic uptake of lactate during intense exercise in humans. AB - We evaluated whether the increase in blood lactate with intense exercise is influenced by a low hepatosplanchnic blood flow as assessed by indocyanine green dye elimination and blood sampling from an artery and the hepatic vein in eight men. The hepatosplanchnic blood flow decreased from a resting value of 1.6 +/- 0.1 to 0.7 +/- 0.1 (SE) l/min during exercise. Yet the hepatosplanchnic O2 uptake increased from 67 +/- 3 to 93 +/- 13 ml/min, and the output of glucose increased from 1.1 +/- 0.1 to 2.1 +/- 0.3 mmol/min (P < 0.05). Even at the lowest hepatosplanchnic venous hemoglobin O2 saturation during exercise of 6%, the average concentration of glucose in arterial blood was maintained close to the resting level (5.2 +/- 0.2 vs. 5.5 +/- 0.2 mmol/l), whereas the difference between arterial and hepatic venous blood glucose increased to a maximum of 22 mmol/l. In arterial blood, the concentration of lactate increased from 1.1 +/- 0.2 to 6.0 +/- 1.0 mmol/l, and the hepatosplanchnic uptake of lactate was elevated from 0.4 +/- 0.06 to 1.0 +/- 0.05 mmol/min during exercise (P < 0.05). However, when the hepatosplanchnic venous hemoglobin O2 saturation became low, the arterial and hepatosplanchnic venous blood lactate difference approached zero. Even with a marked reduction in its blood flow, exercise did not challenge the ability of the liver to maintain blood glucose homeostasis. However, it appeared that the contribution of the Cori cycle decreased, and the accumulation of lactate in blood became influenced by the reduced hepatosplanchnic blood flow. PMID- 11896038 TI - Acetaminophen does not affect 24-h body temperature or sleep in the luteal phase of the menstrual cycle. AB - Body temperature and sleep change in association with increased progesterone in the luteal phase of the menstrual cycle in young women. The mechanism by which progesterone raises body temperature is not known but may involve prostaglandins, inducing a thermoregulatory adjustment similar to that of fever. Prostaglandins also are involved in sleep regulation and potentially could mediate changes in sleep during the menstrual cycle. We investigated the possible role of central prostaglandins in mediating menstrual-associated 24-h temperature and sleep changes by inhibiting prostaglandin synthesis with a therapeutic dose of the centrally acting cyclooxygenase inhibitor acetaminophen in the luteal and follicular phases of the menstrual cycle in young women. Body temperature was raised, and nocturnal amplitude was blunted, in the luteal phase compared with the follicular phase. Acetaminophen had no effect on the body temperature profile in either menstrual cycle phase. Prostaglandins, therefore, are unlikely to mediate the upward shift of body temperature in the luteal phase. Sleep changed during the menstrual cycle: on the placebo night in the luteal phase the women had less rapid eye movement sleep and more slow-wave sleep than in the follicular phase. Acetaminophen did not alter sleep architecture or subjective sleep quality. Prostaglandin inhibition with acetaminophen, therefore, had no effect on the increase in body temperature or on sleep in the midluteal phase of the menstrual cycle in young women, making it unlikely that central prostaglandin synthesis underlies these luteal events. PMID- 11896039 TI - Fetal and adult cerebral artery K(ATP) and K(Ca) channel responses to long-term hypoxia. AB - High-altitude long-term hypoxia (LTH) alters cerebral vascular contractile and relaxation responses in both fetus and adult. We tested the hypotheses that LTH mediated vascular responses were secondary to altered K+ channel function and that in the fetus these responses differ from those of the adult. In middle cerebral arteries (MCA) from both nonpregnant adult and fetal (approximately 140 days gestation) sheep, which were either acclimatized to high altitude (3,820 m) or sea-level controls, we measured norepinephrine (NE)-induced contractions and intracellular Ca2+ concentration ([Ca2+]i) simultaneously, in the presence or absence of different K+ channel openers or blockers. In adult MCA, LTH was associated with approximately 20% decrease in NE-induced tension and [Ca2+]i, with a significant increase in Ca2+ sensitivity. In contrast, in fetal MCA, LTH failed to affect significantly NE-induced contraction or [Ca2+]i but significantly decreased the ATP-sensitive K+ (K(ATP)) channel and Ca2+-activated K+ (K(Ca)) channel-mediated relaxation. The significant effect of K(ATP) and K(Ca) channel activators on the relaxation responses and the fact that K+ channels play a key role in myogenic tone support the hypotheses that K+ channels play an important role in hypoxia-mediated responses. These results also support the hypothesis of significant developmental differences with maturation from fetus to adult. PMID- 11896040 TI - Estimate of the subepithelial hydrostatic pressure that drives inflammatory transudate into the airway lumen. AB - Inflammatory diseases of the upper respiratory tract are characterized by flow of plasma filtrate across the epithelium into the airway lumen ("transudation"). Elsewhere, we have proposed that extravasation from microvessels causes edema, and this is associated with elevated subepithelial hydrostatic pressure that drives transudation. To test this hypothesis, we have attempted to block transudation by elevating luminal hydrostatic pressure. We measured the appearance of plasma markers into the lumen of an isolated perfused segment of rat trachea in vivo and found that stimulation of one vagal nerve caused a rapid (half-time approximately 5 min) and nonselective increase in the flow of markers from blood to airway lumen. Leukocyte migration also caused transudation that developed much more slowly (half-time = 2-3 h). In both cases, transudation was blocked by application of luminal hydrostatic pressures. The critical luminal pressure needed to block vagally induced transudation was approximately 4.5 cmH2O, and, to block epithelial transudation induced by leukocyte traffic, it was 3 cmH2O, and we conclude that these are the subepithelial pressures that drive inflammatory transudation into the airway lumen. PMID- 11896041 TI - Regional measurement of canine skeletal muscle blood flow by positron emission tomography with H2(15)O. AB - Positron emission tomography (PET) with H2(15)O was used as an in vivo, relatively noninvasive, quantitative method for measuring regional blood flow to hindlimb skeletal muscle of anesthetized dogs. A hydrooccluder positioned on the femoral artery was used to reduce flow, and high-flow states were produced by local infusion of adenosine. Three to four measurements were made in each animal. Approximately 40 mCi of H2(15)O were injected intravenously, and serial images and arterial blood samples were acquired over 2.5 min. Data analysis was performed by fitting tissue and arterial blood time-activity curves to a modified, single-compartment Kety model. The model equation was also solved on a pixel-by-pixel basis to yield maps of regional skeletal muscle blood flow. After each PET determination, flow was measured with radioactive microspheres. Results of the PET measurements demonstrated that basal flow to hindlimb skeletal muscle was 3.83 +/- 0.36 ml x min(-1) x 100 g(-1) (mean +/- SE). This value was in excellent agreement with the microsphere data, 3.73 +/- 0.32 ml x min(-1) x 100 g(-1) (P = 0.69, not significant). Adenosine infusion resulted in flows as high as 30 ml x min(-1) x 100 g(-1), and the PET and microsphere data were highly correlated over the entire range of flows (r2 = 0.98, P < 0.0001). We conclude that muscle blood flow can be accurately measured in vivo by PET with H2(15)O and that this approach offers promise for application in human studies of muscle metabolism under varying pathophysiological states. PMID- 11896042 TI - Some historical perspectives on thermoregulation. AB - In this paper, selected historical aspects of thermoregulation and fever are presented as background to the application of molecular biology to thermoregulation. Temperature-sensing mechanisms, coordination of thermal information, thermoregulatory circuitry, efferent responses to thermal stimuli, set point mechanisms, and some of the mechanisms and consequences of fever and hyperthermia are highlighted. Neurotransmitters used in thermoregulatory circuits are also discussed. An attempt is made to include information from comparative physiological sources. Possible future avenues of research in the light of recent new technologies are also presented. PMID- 11896043 TI - Invited review: Effects of heat and cold stress on mammalian gene expression. AB - This review examines the effects of thermal stress on gene expression, with special emphasis on changes in the expression of genes other than heat shock proteins (HSPs). There are approximately 50 genes not traditionally considered to be HSPs that have been shown, by conventional techniques, to change expression as a result of heat stress, and there are <20 genes (including HSPs) that have been shown to be affected by cold. These numbers will likely become much larger as gene chip array and proteomic technologies are applied to the study of the cell stress response. Several mechanisms have been identified by which gene expression may be altered by heat and cold stress. The similarities and differences between the cellular responses to heat and cold may yield key insights into how cells, and by extension tissues and organisms, survive and adapt to stress. PMID- 11896044 TI - Invited review: Interplay between molecular chaperones and signaling pathways in survival of heat shock. AB - Heat shock of mammalian cells causes protein damage and activates a number of signaling pathways. Some of these pathways enhance the ability of cells to survive heat shock, e.g., induction of molecular chaperones [heat shock protein (HSP) HSP72 and HSP27], activation of the protein kinases extracellular signal regulated kinase and Akt, and phosphorylation of HSP27. On the other hand, heat shock can activate a stress kinase, c-Jun NH2-terminal kinase, thus triggering both apoptotic and nonapoptotic cell death programs. Recent data indicate that kinases activated by heat shock can regulate synthesis and functioning of the molecular chaperones, and these chaperones modulate activity of the cell death and survival pathways. Therefore, the overall balance of the pathways and their interplay determine whether a cell exposed to heat shock will die or survive and become stress tolerant. PMID- 11896046 TI - Selected contribution: Hyperthermia-induced intestinal permeability and the role of oxidative and nitrosative stress. AB - The purpose of this study was to characterize intestinal permeability changes over a range of physiologically relevant body temperatures in vivo and in vitro. Initially, FITC-dextran (4,000 Da), a large fluorescent molecule, was loaded into the small intestine of anesthetized rats. The rats were then maintained at approximately 37 degrees C or heated over 90 min to a core body temperature of approximately 41, approximately 41.5, or approximately 42.5 degrees C. Permeability was greater in the 42.5 degrees C group compared with the 37, 41, or 41.5 degrees C groups. Histological analysis revealed intestinal epithelial damage in heated groups. Everted intestinal sacs were then used to further characterize hyperthermia-induced intestinal permeability and to study the potential role of oxidative and nitrosative stress. Increased permeability to 4,000-Da FITC-dextran in both small intestinal and colonic sacs was observed at a temperature of 41.5-42 degrees C compared with 37 degrees C, along with widespread intestinal epithelial damage. Administration of antioxidant enzyme mimics or a nitric oxide synthase inhibitor did not reduce permeability due to heat stress, and tissue concentrations of a lipid peroxidation product were not altered by heat stress, suggesting that oxidative and nitrosative stress were not likely mediators of this phenomenon in vitro. In conclusion, hyperthermia produced increased permeability and marked intestinal epithelial damage both in vivo and in vitro, suggesting that thermal disruption of epithelial membranes contributes to the intestinal barrier dysfunction manifested with heat stress. PMID- 11896049 TI - Structure of apolipophorin-III in discoidal lipoproteins. Interhelical distances in the lipid-bound state and conformational change upon binding to lipid. AB - The structure of apolipophorin III in the lipid-bound state and the extent of the conformational change that takes place when the five-helix bundle apolipoprotein binds to a lipoprotein lipid surface were investigated by fluorescence resonance energy transfer in discoidal lipoproteins. Four intramolecular interhelical distances between helix pairs 1-4, 2-4, 3-4, and 5-4 were estimated by fluorescence resonance energy transfer in both the lipid-free and the lipid-bound states. Depending on the helices pairs, the intramolecular interhelical distances increased between 15 and > or = 20 A upon binding of the apolipoprotein to lipid, demonstrating for the first time that binding to lipid is accompanied by a major change in interhelical distances. Using discoidal lipoproteins made with a combination of apolipophorin III molecules containing donor and acceptor groups and apolipophorin III molecules containing neither donor nor acceptor groups, it was possible to obtain information about intermolecular interhelical distances between the helix 4 of one apolipoprotein and the helices 1, 2, 3, and 5 of a second apolipoprotein residing in the same discoidal lipoprotein. Altogether, the estimated intermolecular and intramolecular interhelical distances suggest a model in which the apolipoprotein arranges in pairs of antiparallel and fully extended polypeptide chains surrounding the periphery of the bilayer disc. PMID- 11896047 TI - Selected contribution: Differential expression of stress-related genes with aging and hyperthermia. AB - Aging is associated with a reduced capacity to cope with physiological stress. To study the molecular mechanisms associated with the decline in stress tolerance that accompanies aging, differences in gene expression between young and old Fischer 344 rats under euthermic control conditions or in response to hyperthermic challenge were evaluated using a cDNA array containing 207 stress related genes. In the nonstressed control condition, aging resulted in selective upregulation of stress protein genes and transcripts involved in cell growth, death, and signaling, along with a downregulation of genes involved in antioxidant defenses and drug metabolism. Heat stress resulted in a broad induction of genes in the antioxidant and drug metabolism categories and transcripts involved in DNA, RNA, and protein synthesis for both age groups. Old animals had a robust upregulation of genes involved in cell growth, death, and signaling after heat challenge, along with a blunted expression of stress response genes. In contrast, young animals had a strong induction of stress response genes after hyperthermic challenge. Changes in expression of selected genes were confirmed by RT-PCR analysis. These findings suggest that aging results in altered gene expression in response to heat stress that is indicative of decreased stress protein transcription and increased expression of oxidative stress-related genes. Thus our findings support the postulate that transcriptional changes in response to a physiological challenge such as hyperthermia contribute to the loss of stress tolerance in older organisms. PMID- 11896050 TI - Rat mast cell protease 4 is a beta-chymase with unusually stringent substrate recognition profile. AB - Activated mast cells release a variety of potent inflammatory mediators including histamine, cytokines, proteoglycans, and serine proteases. The serine proteases belong to either the chymase (chymotrypsin-like substrate specificity) or tryptase (trypsin-like specificity) family. In this report we have investigated the substrate specificity of a recently identified mast cell protease, rat mast cell protease-4 (rMCP-4). Based on structural homology, rMCP-4 is predicted to belong to the chymase family, although rMCP-4 has previously not been characterized at the protein level. rMCP-4 was expressed with an N-terminal His tag followed by an enterokinase site substituting for the native activation peptide. The enterokinase-cleaved fusion protein was labeled by diisopropyl fluorophosphate, demonstrating that it is an active serine protease. Moreover, rMCP-4 hydrolyzed MeO-Suc-Arg-Ala-Tyr-pNA, thus verifying that this protease belongs to the chymase family. rMCP-4 bound to heparin, and the enzymatic activity toward MeO-Suc-Arg-Ala-Tyr-pNA was strongly enhanced in the presence of heparin. Detailed analysis of the substrate specificity was performed using peptide phage display technique. After six rounds of amplification a consensus sequence, Leu-Val-Trp-Phe-Arg-Gly, was obtained. The corresponding peptide was synthesized, and rMCP-4 was shown to cleave only the Phe-Arg bond in this peptide. This demonstrates that rMCP-4 displays a striking preference for bulky/aromatic amino acid residues in both the P1 and P2 positions. PMID- 11896048 TI - Characterization of a brain-enriched chaperone, MRJ, that inhibits Huntingtin aggregation and toxicity independently. AB - Molecular chaperones are involved in a wide range of cellular events, such as protein folding and oligomeric protein complex assembly. DnaK- and DnaJ-like proteins are the two major classes of molecular chaperones in mammals. Recent studies have shown that DnaJ-like family proteins can inhibit polyglutamine aggregation, a hallmark of many neurodegenerative diseases, including Huntington's disease (HD). Although most DnaJ-like proteins studied are ubiquitously expressed, some have restricted expression, so it is possible that some specific chaperones may affect polyglutamine aggregation in specific neurons. In this report, we describe the isolation of a DnaJ-like protein MRJ and the characterization of its chaperone activity. Tissue distribution studies showed that MRJ is highly enriched in the central nervous system. In an in vitro cell model of HD, overexpressed MRJ effectively suppressed polyglutamine dependent protein aggregation, caspase activity, and cellular toxicity. Collectively, these results suggest that MRJ has a relevant functional role in neurons. PMID- 11896051 TI - Heterodimerization of somatostatin and opioid receptors cross-modulates phosphorylation, internalization, and desensitization. AB - Heterodimerization has been shown to modulate the ligand binding, signaling, and trafficking properties of G protein-coupled receptors. However, to what extent heterodimerization may alter agonist-induced phosphorylation and desensitization of these receptors has not been documented. We have recently shown that heterodimerization of sst(2A) and sst(3) somatostatin receptors results in inactivation of sst(3) receptor function (Pfeiffer, M., Koch, T., Schroder, H., Klutzny, M., Kirscht, S., Kreienkamp, H. J., Hollt, V., and Schulz, S. (2001) J. Biol. Chem. 276, 14027-14036). Here we examine dimerization of the sst(2A) somatostatin receptor and the mu-opioid receptor, members of closely related G protein-coupled receptor families. In coimmunoprecipitation studies using differentially epitope-tagged receptors, we provide direct evidence for heterodimerization of sst(2A) and MOR1 in human embryonic kidney 293 cells. Unlike heteromeric assembly of sst(2A) and sst(3), sst(2A)-MOR1 heterodimerization did not substantially alter the ligand binding or coupling properties of these receptors. However, exposure of the sst(2A)-MOR1 heterodimer to the sst(2A)-selective ligand L-779,976 induced phosphorylation, internalization, and desensitization of sst(2A) as well as MOR1. Similarly, exposure of the sst(2A)-MOR1 heterodimer to the mu-selective ligand [d-Ala(2),Me Phe(4),Gly(5)-ol]enkephalin induced phosphorylation and desensitization of both MOR1 and sst(2A) but not internalization of sst(2A). Cross-phosphorylation and cross-desensitization of the sst(2A)-MOR1 heterodimer were selective; they were neither observed with the sst(2A)-sst(3) heterodimer nor with the endogenously expressed lysophosphatidic acid receptor. Heterodimerization may thus represent a novel regulatory mechanism that could either restrict or enhance phosphorylation and desensitization of G protein-coupled receptors. PMID- 11896052 TI - Different residues in the GABA(A) receptor alpha 1T60-alpha 1K70 region mediate GABA and SR-95531 actions. AB - Although gamma-aminobutyric acid type A receptor agonists and antagonists bind to a common site, they produce different conformational changes within the site because agonists cause channel opening and antagonists do not. We used the substituted cysteine accessibility method and two-electrode voltage clamping to identify residues within the binding pocket that are important for mediating these different actions. Each residue from alpha(1)T60 to alpha(1)K70 was mutated to cysteine and expressed with wild-type beta(2) subunits in Xenopus oocytes. Methanethiosulfonate reagents reacted with alpha(1)T60C, alpha(1)D62C, alpha(1)F64C, alpha(1)R66C, alpha(1)S68C, and alpha(1)K70C. gamma-Aminobutyric acid (GABA) slowed methanethiosulfonate modification of alpha(1)F64C, alpha(1)R66C, and alpha(1)S68C, whereas SR-95531 slowed modification of alpha(1)D62C, alpha(1)F64C, and alpha(1)R66C, demonstrating that different residues are important for mediating GABA and SR-95531 actions. In addition, methanethiosulfonate reaction rates were fastest for alpha(1)F64C and alpha(1)R66C, indicating that these residues are located in an open, aqueous environment lining the core of the binding pocket. Positively charged methanethiosulfonate reagents derivatized alpha(1)F64C and alpha(1)R66C significantly faster than a negatively charged reagent, suggesting that a negative subsite important for interacting with the ammonium group of GABA exists within the binding pocket. Pentobarbital activation of the receptor increased the rate of methanethiosulfonate modification of alpha(1)D62C and alpha(1)S68C, demonstrating that parts of the binding site undergo structural rearrangements during channel gating. PMID- 11896053 TI - Rac activation induces NADPH oxidase activity in transgenic COSphox cells, and the level of superoxide production is exchange factor-dependent. AB - Transient expression of constitutively active Rac1 derivatives, (G12V) or (Q61L), was sufficient to induce phagocyte NADPH oxidase activity in a COS-7 cell model in which human cDNAs for essential oxidase components, gp91(phox), p22(phox), p47(phox), and p67(phox), were expressed as stable transgenes. Expression of constitutively active Rac1 in "COS(phox)" cells induced translocation of p47(phox) and p67(phox) to the membrane. Furthermore, translocation of p47(phox) was induced in the absence of p67(phox) expression, even though Rac does not directly bind p47(phox). Rac effector domain point substitutions (A27K, G30S, D38A, Y40C), which can selectively eliminate interaction with different effector proteins, impaired Rac1V12-induced superoxide production. Activation of endogenous Rac1 by expression of constitutively active Rac-guanine nucleotide exchange factor (GEF) derivatives was sufficient to induce high level NADPH oxidase activity in COS(phox) cells. The constitutively active form of the hematopoietic-specific GEF, Vav1, was the most effective at activating superoxide production, despite detection of higher levels of Rac1-GTP upon expression of constitutively active Vav2 or Tiam1 derivatives. These data suggest that Rac can play a dual role in NADPH oxidase activation, both by directly participating in the oxidase complex and by activating signaling events leading to oxidase assembly, and that Vav1 may be the physiologically relevant GEF responsible for activating this Rac-regulated complex. PMID- 11896054 TI - X-ray structure of a serine protease acyl-enzyme complex at 0.95-A resolution. AB - Kinetic analyses led to the discovery that N-acetylated tripeptides with polar residues at P3 are inhibitors of porcine pancreatic elastase (PPE) that form unusually stable acyl-enzyme complexes. Peptides terminating in a C-terminal carboxylate were more potent than those terminating in a C-terminal amide, suggesting recognition by the oxy-anion hole is important in binding. X-ray diffraction data were recorded to 0.95-A resolution for an acyl-enzyme complex formed between PPE and N-acetyl-Asn-Pro-Ile-CO2H at approximately pH 5. The accuracy of the crystallographic coordinates allows structural issues concerning the mechanism of serine proteases to be addressed. Significantly, the ester bond of the acyl-enzyme showed a high level of planarity, suggesting geometric strain of the ester link is not important during catalysis. Several hydrogen atoms could be clearly identified and were included within the model. In keeping with a recent x-ray structure of subtilisin at 0.78 A (1), limited electron density is visible consistent with the putative location of a hydrogen atom approximately equidistant between the histidine and aspartate residues of the catalytic triad. Comparison of this high resolution crystal structure of the acyl-enzyme complex with that of native elastase at 1.1 A (2) showed that binding of the N-terminal part of the substrate can be accommodated with negligible structural rearrangements. In contrast, comparison with structures obtained as part of "time resolved" studies on the reacting acyl-enzyme complex at >pH 7 (3) indicate small but significant structural differences, consistent with the proposed synchronization of ester hydrolysis and substrate release. PMID- 11896055 TI - ERK negatively regulates the epidermal growth factor-mediated interaction of Gab1 and the phosphatidylinositol 3-kinase. AB - We have examined the ability of epidermal growth factor (EGF)-stimulated ERK activation to regulate Grb2-associated binder-1 (Gab1)/phosphatidylinositol 3 kinase (PI3K) interactions. Inhibiting ERK activation with the MEK inhibitor U0126 increased the EGF-stimulated association of Gab1 with either full-length glutathione S-transferase-p85 or the p85 C-terminal Src homology 2 (SH2) domain, a result reproduced by co-immunoprecipitation of the native proteins from intact cells. This increased association of Gab1 and the PI3K correlates with an increase in PI3K activity and greater phosphorylation of Akt. This result is in direct contrast to what we have previously reported following HGF stimulation where MEK inhibition decreased the HGF-stimulated association of Gab1 and p85. In support of this divergent effect of ERK on Gab1/PI3K association following HGF and EGF stimulation, U0126 decreased the HGF-stimulated association of p85 and the Gab1 c-Met binding domain but did not alter the EGF-stimulated association of p85 and the c-Met binding domain. An examination of the mechanism of this effect revealed that the treatment of cells with EGF + U0126 increased the tyrosine phosphorylation of Gab1 as well as its association with another SH2-containing protein, SHP2. Furthermore, overexpression of a catalytically inactive form of SHP2 or pretreatment with pervanadate markedly increased EGF-stimulated Gab1 tyrosine phosphorylation. These experiments demonstrate that EGF and HGF-mediated ERK activation result in divergent effects on Gab1/PI3K signaling. HGF-stimulated ERK activation increases the Gab1/PI3K association, whereas EGF-stimulated ERK activation results in a decrease in the tyrosine phosphorylation of Gab1 and a decreased association with the PI3K. SHP2 is shown to associate with and dephosphorylate Gab1, suggesting that EGF-stimulated ERK might act through the regulation of SHP2. PMID- 11896056 TI - Molecular and functional characterization of a murine calcium-activated chloride channel expressed in smooth muscle. AB - To identify the gene products responsible for the calcium-activated chloride current in smooth muscle, reverse transcription-PCR with degenerate primers was performed on mouse intestine and other organs. A new member of the CLCA gene family was identified, mCLCA4, that is expressed preferentially in organs containing a high percentage of smooth muscle cells, including intestine, stomach, uterus, bladder, and aorta. Reverse transcription-PCR using template RNA prepared from mouse bladder and stomach smooth muscle layers dissected free of mucosa yielded mCLCA4-specific bands. In situ hybridization with an mCLCA4 specific probe confirmed prominent expression in smooth muscle of major vessels of the heart but not cardiac muscle. High expression was also detected in the gastrointestinal tract, in bronchioles, and in aortic and lung endothelial cells. Transient expression of mCLCA4 in 293T cells resulted in the appearance of a prominent calcium-activated chloride current. Whole-cell currents activated by ionomycin or methacholine were anion-selective and showed minimal rectification or voltage-dependent gating. Similar to endogenous currents in smooth muscle cells, methacholine-induced currents were transient, and spontaneous transient inward currents were occasionally observed at resting membrane potentials. These results link calcium-activated chloride channels in smooth muscle with a gene family whose members have been implicated in cystic fibrosis, cancer, and asthma. PMID- 11896057 TI - In vitro evidence that the untranslated leader of the HIV-1 genome is an RNA checkpoint that regulates multiple functions through conformational changes. AB - The HIV-1 RNA genome forms dimers through base pairing of a palindromic 6-mer sequence that is exposed in the loop of the dimer initiation signal (DIS) hairpin structure (loop-loop kissing). The HIV-1 leader RNA can adopt a secondary structure conformation that is not able to dimerize because the DIS hairpin is not folded. Instead, this DIS motif is base-paired in a long distance interaction (LDI) that extends the stem of the primer-binding site domain. In this study, we show that targeting of the LDI by either antisense oligonucleotides or specific mutations can induce the conformational switch to a branched multiple hairpin (BMH) structure, and this LDI-to-BMH switch coincides with increased RNA dimerization. Another interesting finding is that the extended LDI stem can resist a certain level of destabilization, indicating that a buffer is created to prevent a premature conformational switch and early dimerization. Because the tRNA(Lys3) primer for reverse transcription anneals to multiple sequence elements of the HIV-1 leader RNA, including sequences in the LDI stem, we tested whether tRNA-annealing can destabilize the LDI stem such that RNA dimerization is triggered. Using a combination of stem-destabilizing approaches, we indeed measured a small but significant effect of tRNA-annealing on the ability of the RNA template to form dimers. This observation suggests that HIV-1 RNA can act as a checkpoint to control and coordinate different leader functions through conformational switches. This in vitro result should be verified in subsequent in vivo studies with HIV-infected cells. PMID- 11896058 TI - Conformational analysis of the androgen receptor amino-terminal domain involved in transactivation. Influence of structure-stabilizing solutes and protein protein interactions. AB - The androgen receptor (AR) is a member of the nuclear receptor superfamily. Sequences within the large amino-terminal domain of the receptor have been shown to be important for transactivation and protein-protein interactions; however, little is known about the structure and folding of this region. In the present study we show that a 344-amino acid polypeptide representing the main determinants for transactivation has the propensity to form alpha-helical structure and that mutations which disrupt putative helical regions alter conformation. Folding of the AR was observed in the presence of the helix stabilizing solvent trifluoroethanol and the natural osmolyte trimethylamine N oxide (TMAO). TMAO resulted in the movement of two tryptophan residues to a less solvent-exposed environment and the formation of secondary/tertiary structure resistant to protease cleavage. Critically, binding to the RAP74 subunit of the general transcription factor TFIIF resulted in extensive protease resistance, consistent with induced folding of the receptor transactivation domain. These data indicate that this region of the AR is structurally flexible and folds into a stable conformation upon interactions with a component of the general transcription machinery. PMID- 11896059 TI - Identification of amino acids important for the catalytic activity of the collagen glucosyltransferase associated with the multifunctional lysyl hydroxylase 3 (LH3). AB - Collagen glucosyltransferase (GGT) activity has recently been shown to be associated with human lysyl hydroxylase (LH) isoform 3 (LH3) (Heikkinen, J., Risteli, M., Wang, C., Latvala, J., Rossi, M., Valtavaara, M., Myllyla, R. (2000) J. Biol. Chem. 275, 36158-36163). The LH and GGT activities of the multifunctional LH3 protein modify lysyl residues in collagens posttranslationally to form hydroxylysyl and glucosylgalactosyl hydroxylysyl residues respectively. We now report that in the nematode, Caenorhabditis elegans, where only one ortholog is found for lysyl hydroxylase, the LH and GGT activities are also associated with the same gene product. The aim of the present studies is the identification of amino acids important for the catalytic activity of GGT. Our data indicate that the GGT active site is separate from the carboxyl terminal LH active site of human LH3, the amino acids important for the GGT activity being located at the amino-terminal part of the molecule. Site-directed mutagenesis of a conserved cysteine at position 144 to isoleucine and a leucine at position 208 to isoleucine caused a marked reduction in GGT activity. These amino acids were conserved in C. elegans LH and mammalian LH3, but not in LH1 or LH2, which lack GGT activity. The data also reveal a DXD-like motif in LH3 characteristic of many glycosyltransferases and the mutagenesis of aspartates of this motif eliminated the GGT activity. Reduction in GGT activity was not accompanied by a change in the LH activity of the molecule. Thus GGT activity can be manipulated independently of LH activity in LH3. These data provide the information needed to design knock-out studies for investigation of the function of glucosylgalactosyl hydroxylysyl residues of collagens in vivo. PMID- 11896060 TI - Molecular determinants of the sensory and motor neuron-derived factor insertion into plasma membrane. AB - The sensory and motor neuron-derived factor (SMDF) is a type III neuregulin that regulates development and proliferation of Schwann cells. Although SMDF has been shown to be a type II protein, the molecular determinants of membrane biogenesis, insertion, and topology remain elusive. Here we used heterologous expression of a yellow fluorescent protein-SMDF fusion protein along with a stepwise deletion strategy to show that the apolar/uncharged segment (Ile(76)-Val(100)) acts as an internal, uncleaved membrane insertion signal that defines the topology of the protein. Unexpectedly, removal of the transmembrane segment (TM) did not eliminate completely membrane association of C-terminal fragments. TM-deleted fusion proteins, bearing the amino acid segment (Ser(283)-Glu(296)) located downstream to the epidermal growth factor-like motif, strongly interacted with plasma membrane fractions. However, synthetic peptides patterned after this segment did not insert into artificial lipid vesicles, suggesting that membrane interaction of the SMDF C terminus may be the result of a post-translational modification. Subcellular localization studies demonstrated that the 40-kDa form, but not the 83-kDa form, of SMDF was segregated into lipid rafts. Deletion of the N-terminal TM did not affect the interaction of the protein with these lipid microdomains. In contrast, association with membrane rafts was abolished completely by truncation of the protein C terminus. Collectively, these findings are consistent with a topological model for SMDF in which the protein associates with the plasma membrane through both the TM and the C-terminal end domains resembling the topology of other type III neuregulins. The TM defines its characteristic type II membrane topology, whereas the C terminus is a newly recognized anchoring motif that determines its compartmentalization into lipid rafts. The differential localization of the 40- and 83-kDa forms of the neuregulin into rafts and non-raft domains implies a central role in the protein biological activity. PMID- 11896061 TI - Association of the human SUMO-1 protease SENP2 with the nuclear pore. AB - SUMO-1 is a small ubiquitin-like protein that can be covalently conjugated to other proteins. A family of proteases catalyzes deconjugation of SUMO-1 containing species. Members of this family also process newly synthesized SUMO-1 into its conjugatable form. To understand these enzymes better, we have examined the localization and behavior of the human SUMO-1 protease SENP2. Here we have shown that SENP2 associates with the nuclear face of nuclear pores and that this association requires protein sequences near the N terminus of SENP2. We have also shown that SENP2 binds to Nup153, a nucleoporin that is localized to the nucleoplasmic face of the pore. Nup153 binding requires the same domain of SENP2 that mediates its targeting in vivo. Removal of the Nup153-interacting region of SENP2 results in a significant change in the spectrum of SUMO-1 conjugates within the cell. Our results suggest that association with the pore plays an important negative role in the regulation of SENP2, perhaps by restricting its activity to a subset of the conjugated proteins within the nucleus. PMID- 11896062 TI - A prenylated p67phox-Rac1 chimera elicits NADPH-dependent superoxide production by phagocyte membranes in the absence of an activator and of p47phox: conversion of a pagan NADPH oxidase to monotheism. AB - Activation of the superoxide-generating NADPH oxidase of phagocytes is the result of the assembly of a membrane-localized flavocytochrome (cytochrome b(559)) with the cytosolic components p47(phox), p67(phox), and the small GTPase Rac. Activation can be reproduced in an in vitro system in which cytochrome b(559) containing membranes are mixed with cytosolic components in the presence of an anionic amphiphile. We proposed that the essential event in activation is the interaction between p67(phox) and cytochrome b(559) and that Rac and p47(phox) serve as carriers for p67(phox) to the membrane. When prenylated, Rac can fulfill the carrier function by itself, supporting oxidase activation by p67(phox) in the absence of p47(phox) and amphiphile. We now show that a single chimeric protein, consisting of residues 1-212 of p67(phox) and full-length Rac1 (residues 1-192), prenylated in vitro and exchanged to GTP, becomes a potent oxidase activator in the absence of any other component or stimulus. Oxidase activation by prenylated chimera p67(phox) (1-212)-Rac1 (1-192) is accompanied by its spontaneous association with membranes. Prenylated chimeras p67(phox) (1-212)-Rac1 (178-192) and p67(phox) (1-212)-Rac1 (189-192), containing specific C-terminal regions of Rac1, are inactive; the activity of the first but not of the second chimera can be rescued by supplementation with exogenous nonprenylated Rac1-GTP. An analysis of prenylated p67(phox)-Rac1 chimeras suggests that the basic requirements for oxidase activation are: (i) a "two signals" membrane-localizing motif present in Rac, comprising the prenyl group and a C-terminal polybasic sequence and (ii) an intrachimeric or extrachimeric protein-protein interaction between p67(phox) and Rac1, causing a conformational change in the "activation domain" in p67(phox). PMID- 11896063 TI - Characterization of the matrilin coiled-coil domains reveals seven novel isoforms. AB - Matrilins constitute a family of four oligomeric extracellular proteins that are involved in the development and homeostasis of cartilage and bone. To reveal their homo- and heterotypic oligomerization propensities, we analyzed the four human matrilin coiled-coil domains by biochemical and biophysical methods. These studies not only confirmed the homo- and heterotypic oligomerization states reported for the full-length proteins but revealed seven novel matrilin isoforms. Specific heterotrimeric interactions of variable chain stoichiometries were observed between matrilin-1 and matrilin-2, matrilin-1 and matrilin-4, and matrilin-2 and matrilin-4. In addition, matrilin-1 formed two different specific heterotetramers with matrilin-3. Interestingly, a distinct heterotrimer consisting of three different chains was formed between matrilin-1, matrilin-2, and matrilin-4. No interactions, however, were observed between matrilin-2 and matrilin-3 or between matrilin-3 and matrilin-4. Both homo- and heterotypic oligomers folded into parallel disulfide-linked structures, although coiled-coil formation was not dependent on disulfide bridge formation. Our results indicate that the heterotypic preferences seen for the matrilin coiled-coil domains are the result of the packing of the hydrophobic core rather than ionic interactions. Mass spectrometry revealed that the concentrations of the individual chains statistically determined the stoichiometry of the heteromers, suggesting that formation of the different matrillin chain combinations is controlled by expression levels. PMID- 11896066 TI - Clinical governance and pathology. AB - This article looks at clinical governance and pathology. Clinical governance should be an important tool in seeking quality improvement within the National Health Service. But how as pathologists should we go about it? PMID- 11896065 TI - Hodgkin's lymphoma: the pathologist's viewpoint. AB - Despite its well known histological and clinical features, Hodgkin's lymphoma (HL) has recently been the object of intense research activity, leading to a better understanding of its phenotype, molecular characteristics, histogenesis, and possible mechanisms of lymphomagenesis. There is complete consensus on the B cell derivation of the tumour in most cases, and on the relevance of Epstein-Barr virus infection and defective cytokinesis in at least a proportion of patients. The REAL/WHO classification recognises a basic distinction between lymphocyte predominance HL (LP-HL) and classic HL (CHL), reflecting the differences in clinical presentation and behaviour, morphology, phenotype, and molecular features. CHL has been classified into four subtypes: lymphocyte rich, nodular sclerosing, with mixed cellularity, and lymphocyte depleted. The borders between CHL and anaplastic large cell lymphoma have become sharper, whereas those between LP-HL and T cell rich B cell lymphoma remain ill defined. Treatments adjusted to the pathobiological characteristics of the tumour in at risk patients have been proposed and are on the way to being applied. PMID- 11896067 TI - New additions to antibody panels in the characterisation of chronic lymphoproliferative disorders. AB - Advances in flow cytometry techniques and the availability of monoclonal antibodies that detect key functional molecules on lymphocytes have contributed greatly to a more precise diagnosis of the chronic lymphoproliferative disorders. In addition to the diagnostic value, the expression of certain markers such as p53 or CD38 provides relevant prognostic information to the clinician. Beyond their diagnostic and prognostic value, immunological markers play a major role in the detection of minimal residual disease, enabling the clinician to estimate more accurately the response to chemotherapy. Those monoclonal antibodies that are relevant to the characterisation of the chronic lymphoproliferative disorders and that could be incorporated in a routine practice are discussed. PMID- 11896068 TI - Increased nm23 immunoreactivity is associated with selective inhibition of systemic tumour cell dissemination. AB - AIMS: In vitro transfection experiments show that the nm23 gene suppresses metastasis, although the evidence from clinical studies is contradictory. The purpose of this study was to investigate whether nm23 selectively influences systemic, pleural, and lymphatic metastasis in non-small cell lung cancer (NSCLC). METHODS: Forty two patients undergoing resection of NSCLC and lymph node sampling were enrolled prospectively. In each case, a bone marrow aspirate, pleural lavage, and lymph nodes were assessed using immunohistochemistry for epithelial antigens and morphology. The intensity of nm23-H1 immunoreactivity of the primary tumour was compared with the internal control of normal bronchial epithelium in 32 cases where available. The microvessel count (MVC) of each tumour was determined using immunohistochemistry for the endothelial cell marker CD34. RESULTS: Tumour cell dissemination was detected in the bone marrow in 18 patients, in the pleura in seven, and in the lymph nodes in 21. Increased immunoreactivity for nm23 was found in the primary tumour in six patients, with none having tumour cells in the bone marrow, compared with 12 of 26 patients who showed nm23 immunoreactivity equal to or less than the control (Fisher's exact test: p = 0.043). This effect was confirmed to be independent of the MVC on multivariate analysis. There was no significant difference in the incidence of pleural or lymphatic tumour cell dissemination between the two groups. CONCLUSION: nm23 appears to be a suppressor of systemic, but not lymphatic, metastasis in primary NSCLC. PMID- 11896070 TI - Site distribution of oesophagogastric cancer. AB - AIMS: It has been suggested that adenocarcinomas of the lower oesophagus and gastric cardia should be reclassified as oesophagogastric junction (OGJ) cancers. This study aimed to define the frequency of OGJ cancers in a geographically defined population of 4.3 million people. METHODS: All cases of oesophageal and gastric cancer occurring in 1993 were identified by the North Western Regional Cancer Registry. A total of 1192 hospital case notes were reviewed and a study group of 1067 patients was defined. Tumour involvement was documented at individual subsites in the oesophagus and stomach, allowing for tumour presence in more than one oesophageal/gastric subsite. RESULTS: There were 627 tumours in men and 440 in women. The tumour was confined to the oesophagus in 281 (26.3%) cases and to the stomach in 454 (42.6%) cases. The tumour encroached upon or crossed the OGJ in 332 (31.1%) cases. Overall, tumours involved the cardia, OGJ, or lower oesophagus in 633 (59.3%) cases; in 179 (18.5%) cases the tumour involved the lower oesophagus but not the OGJ, and in another 122 (11.4%) cases the cardia was involved but not the OGJ. CONCLUSIONS: Oesophagogastric cancers in this population predominantly involve the OGJ, lower oesophagus, and/or cardia. PMID- 11896071 TI - Age related expression of Werner's syndrome protein in selected tissues and coexpression of transcription factors. AB - AIMS: Werner's syndrome (WS) is an uncommon autosomal recessive disease resulting from mutational inactivation of human WRN helicase, Werner's syndrome protein (WRNp). Patients with WS progressively develop a variety of aging characteristics after puberty. The aim of this study was to determine the distribution of WRNp and the expression of the transcription factors regulating WRN gene expression in a variety of human organs in an attempt to understand the WS phenotype. METHODS: Tissue specimens were obtained from 16 controls aged from 27 gestational weeks to 70 years of age and a 56 year old female patient with WS. The distribution of WRNp and the expression of the transcription factors regulating WRN gene expression-SP1, AP2, and retinoblastoma protein (Rb)- were studied in the various human organs by immunohistochemical and immunoblot analyses. RESULTS: In the healthy controls after puberty, high expression of WRNp was detected in seminiferous epithelial cells and Leydig cells in the testis, glandular acini in the pancreas, and the zona fasciculata and zona reticularis in the adrenal cortex. In addition, the SP1 and AP2 transcription factors, which regulate WRNp gene expression, appeared in an age dependent manner in those regions where WRNp was expressed. In controls after puberty, SP1 was expressed in the testis and adrenal gland, whereas AP2 was expressed in the pancreas. CONCLUSIONS: These findings suggest that the age specific onset of WS may be related to age dependent expression of WRNp in specific organs. PMID- 11896072 TI - Workload and stress in consultant medical microbiologists and virologists: a questionnaire survey. AB - AIMS: To document demography, changing workload patterns, job satisfaction, morale, and prevalence of stress and psychological morbidity among UK consultant medical microbiologists and virologists. METHOD: A questionnaire survey of all identified UK practising consultant medical microbiologists and virologists (n= 464). RESULTS: Among 367 respondents (79%), there were 33 virologists and at least 89 single handed consultants. Over half the respondents (58%) were working a 1 : 1 or 1 : 2 on call rota during the week and a similar proportion (51%) at weekends. Of all consultants (including those working part time), 56% were working more than 48 hours weekly. Working more than 48 hours weekly, and being on call 1 : 1 or 1 : 2 at weekends, were both independently associated with increased psychological morbidity. Those on call 1 : 1 or 1 : 2 at weekends were also more likely to have low or very low morale. Female consultants were more likely to have higher stress scores. More than half of the respondents (208 of 363; 57%) were making active financial provision to retire early, and 198 of 363 (55%) did not intend to work beyond the age of 60. CONCLUSIONS: The long hours worked by many consultant microbiologists and virologists are in breach of the European Working Time Directive and are associated with a higher degree of psychological morbidity. For most consultants, the frequency of on call commitments is demanding and job satisfaction and morale have deteriorated. Urgent action is needed, particularly to support those working more than 48 hours each week and those on call at weekends 1 : 1 or 1 : 2. However, a major expansion of the consultant establishment cannot be achieved rapidly, and will be slowed further if early retirements become more frequent. PMID- 11896073 TI - Differential expression of cell cycle and apoptosis related proteins in colorectal mucosa, primary colon tumours, and liver metastases. AB - AIMS: Tumour cell growth results from a disturbance in the balance between the rate of proliferation and cell death. In this study, proteins involved in the regulation of cell cycle arrest and apoptosis were studied as possible factors responsible for uncontrolled cell growth in colorectal cancer. METHODS: The expression of proteins involved in these processes was investigated in 48 metastases from patients with colorectal cancer and compared with eight normal colon mucosa samples and 14 primary tumours. Both primary tumours and metastases were obtained from eight patients. The expression of thymidylate synthase (TS), p53, retinoblastoma protein (Rb), Fas receptor, Fas ligand, bcl-2, mcl-1, bax, and bcl-x was measured using immunohistochemistry. Proliferation was determined by Ki67 staining, whereas apoptosis was assessed by M30 immunostaining, which recognises cleaved cytokeratin 18. RESULTS: In the limited number of cases in which paired comparisons were possible, the expression of TS and Ki67 was significantly higher in metastases than in the matched primary tumour samples (p = 0.014 and 0.016, respectively), whereas Rb expression was lower in metastases than in primary tumours (p = 0.024). Fas receptor expression was high in normal mucosa but absent in primary tumours and metastases, whereas the opposite was seen for p53. The expression of bax, mcl-1, and bcl-x in normal mucosa was more apical than that seen in malignant cells, where a more diffuse expression pattern was seen (p < 0.04). Apoptosis was more abundant in primary tumours than in metastases. CONCLUSIONS: These results demonstrate that proliferation and apoptosis are disturbed during colorectal cancer progression, and this is accompanied by loss of Rb and Fas expression, the accumulation of p53 and TS, and changes in the expression patterns of bax, mcl-1, and bcl-xl. PMID- 11896074 TI - Bcl-2 expression and frequency of apoptosis correlate with morphogenesis of colorectal neoplasia. AB - AIM: To investigate whether the expression of apoptosis and cell proliferation related proteins is related to the macroscopic form of colorectal neoplasia. METHODS: The extent of apoptosis, using the 3' end DNA labelling method, and the immunohistochemical expression of cell proliferation (Ki-67) and apoptosis related proteins (Bcl-2, Bak, and p53) were investigated in 64 colorectal adenomas and 22 early carcinomas extending no further than the upper submucosal region. The specimens were classified into three types of macroscopic form (polypoid, flat, and depressed). RESULTS: The Ki-67 labelling index and the Bak score did not differ significantly among each macroscopic form. In contrast, the apoptotic index and the Bcl-2 score changed significantly according to the macroscopic forms. Compared with polypoid and flat tumours, depressed tumours had a significantly lower apoptotic index (2.84, 2.28, and 1.44, respectively) and a significantly lower Bcl-2 score (3.18, 2.70, and 1.64, respectively). The proliferation/apoptosis ratio was significantly lower in polypoid tumours than in the other two macroscopic forms. The Bcl-2 score became significantly lower as the tumours flattened or took on a depressed form. Immunohistochemical p53 overexpression did not correlate with the macroscopic forms. CONCLUSIONS: These results suggest that differences in both Bcl-2 expression and apoptosis may play an important role in the morphogenesis of colorectal neoplasia. PMID- 11896076 TI - Chlamydia pneumoniae in infrequently examined blood vessels. AB - AIM: To determine the prevalence of Chlamydia pneumoniae DNA in infrequently examined blood vessels. METHODS: Vessels obtained from 15 men and six women at coronary artery bypass surgery were tested by a nested polymerase chain reaction (PCR) assay for C pneumoniae DNA. RESULTS: Chlamydia pneumoniae DNA was detected in four of six atheromatous ascending aorta specimens but in none of eight non atheromatous aorta specimens, in six of 11 atheromatous internal mammary artery specimens but in none of seven non-atheromatous internal mammary artery specimens, in five of seven long saphenous vein specimens showing evidence of disease but in none of 12 specimens without evidence of disease, and in two of three previously grafted veins. Overall, C pneumoniae occurred significantly more often in diseased than in normal vessels (p = < 0.00001). CONCLUSIONS: Chlamydia pneumoniae is often present in diseased areas of arteries, including the internal mammary arteries, and even in diseased areas of veins. It is not present in apparently healthy areas of either type of vessel. PMID- 11896077 TI - Extramedullary myeloid cell tumours localised to the mediastinum: a rare clinicopathological entity with unique karyotypic features. AB - Extramedullary myeloid cell tumour (EMMT) localised to the mediastinum is a rare manifestation of acute myeloid leukaemia, forming less than 4% of all cases of EMMT. In contrast to other types of EMMT, cytogenetic characteristics of this rare entity are relatively unknown. This report describes a patient with EMMT who had evidence of superior vena cava syndrome and normal peripheral blood counts at diagnosis. The results from an initial biopsy specimen were consistent with a diagnosis of mediastinal large B cell lymphoma. A diagnosis of acute myeloid leukaemia was made three months after initial diagnosis by bone marrow examination. Review of the initial biopsy specimen showed strong positivity for myeloperoxidase, revealing that the patient had been initially misdiagnosed as having large B cell lymphoma. Cytogenetic studies revealed a near triploid and near tetraploid karyotype with structural abnormalities in 12 and three metaphases, respectively. Review of the literature showed that a near tetraploid or triploid karyotype is found in most of the reported cases of mediastinal EMMT. Thus, the presence of a near triploid/tetraploid karyotype and mediastinal EMMT may represent a specific subset of EMMT. The biological relevance of this observation is discussed. PMID- 11896079 TI - A case of myoepithelial carcinoma displaying biallelic inactivation of the tumour suppressor gene APC in a patient with familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is an autosomal dominant disorder caused by mutation of the APC gene. It is characterised by the appearance of hundreds to thousands of colorectal adenomas in adolescence and the subsequent development of colorectal cancer. Various extracolonic malignancies are associated with FAP, including desmoids and neoplasms of the stomach, duodenum, pancreas, liver, and brain. We present a family affected by FAP with an exon 14 APC mutation displaying two rare extracolonic lesions, a hepatoblastoma and a myoepithelial carcinoma. The hepatoblastoma was found in a male patient aged 2 years. The second lesion, a myoepithelial carcinoma of the right cheek, was found in a female patient aged 14 years. Inactivation of the normal APC allele was demonstrated in this lesion by loss of heterozygosity analysis, thus implicating APC in the initiation or progression of this neoplasm. This is the first reported case of this lesion in a family affected by FAP. PMID- 11896078 TI - Intermediate grade osteosarcoma and chondrosarcoma arising in an osteochondroma. A case report of a patient with hereditary multiple exostoses. AB - A 40 year old man with hereditary multiple exostoses (HME), affecting predominantly his left proximal tibia, distal femur, and proximal femur, underwent resection of an osteochondroma near the trochanter major of his left proximal femur because of malignant transformation of the cartilaginous cap towards secondary peripheral chondrosarcoma. The patient had a history of a papillary thyroid carcinoma four years previously. At examination of the resected specimen, a third malignant tumour, an intermediate grade osteosarcoma (grade II/IV), was found in the osseous stalk of the osteochondroma. Although no mutations were found in the EXT1 and EXT2 genes, the genes involved in HME, or in exons 5-8 of the p53 gene, the development of three malignancies before the age of 40 suggests that this patient is genetically prone to malignant transformation. PMID- 11896080 TI - Seqenenre Taa II, the violent death of a pharaoh. PMID- 11896081 TI - The bioavailability of mupirocin in nasal secretions in vitro. AB - AIM: To determine the bioavailability of mupirocin in human nasal secretions and to assess whether the contents of nasal secretions interact appreciably with this antibiotic. METHODS: The comparative bioavailability of mupirocin and chlorhexidine in nasal secretions was determined by bioassay after one, four, and eight hours of incubation with pooled secretions from three subjects. The interaction of mupirocin with nasal secretions was characterised by matrix assisted laser desorption time of flight mass spectrometry (MALDI-TOF). RESULTS: MALDI-TOF analysis showed that mupirocin was not absorbed by the main fraction of pooled nasal secretions and should remain active. In bioassay, mupirocin retained 100% of its antistaphylococcal activity in nasal secretions, whereas chlorhexidine was significantly reduced from 100 mg/litre to 1.5 mg/litre and from 1000 mg/litre to 38.5 mg/litre, irrespective of incubation time. CONCLUSIONS: The high bioavailability of mupirocin in nasal secretions results from the lack of appreciable molecular interactions. PMID- 11896082 TI - The effect of fixation and processing on the sensitivity of oestrogen receptor assay by immunohistochemistry in breast carcinoma. AB - AIMS: To investigate the effect of different fixation and processing regimens on the assessment of oestrogen receptor (ER) by immunohistochemistry (IHC). METHODS: The ER results for 420 patients from seven different hospitals in which antigen retrieval and IHC were performed centrally were compared. The intensity of ER positivity was assessed semiquantitatively using the Quick score (range 0-7). The scoring profiles of cases from each different source (hospital) were compared to detect differences in the proportion of cases that were negative (Quick score = 0), moderately positive (Quick score = 1-5), and strongly positive (Quick score = 6-7). RESULTS: There were no significant differences (p = 0.3; kappa(2) test) in the proportion of cases in each category. CONCLUSIONS: None of the fixation or processing regimens had a significant adverse effect on the sensitivity of the ER assessment performed by automated immunohistochemistry. PMID- 11896083 TI - Efficient isolation of campylobacters from stools: what are we missing? PMID- 11896084 TI - Evidence for antibiotic induced Clostridium perfringens diarrhoea. PMID- 11896085 TI - Are all hypotheses generated before data analysis prospective? PMID- 11896086 TI - Dear doctor: we really are not sure what dose of capecitabine you should prescribe for your patient. PMID- 11896087 TI - Cooling off hot flashes. PMID- 11896088 TI - Vive la difference: sex and fluorouracil toxicity. PMID- 11896089 TI - Liver metastases from colorectal cancer: the turning point. PMID- 11896090 TI - Unexpected serious toxicity with chemotherapy and antiangiogenic combinations: time to take stock! PMID- 11896091 TI - Effect of soy phytoestrogens on hot flashes in postmenopausal women with breast cancer: a randomized, controlled clinical trial. AB - PURPOSE: Vasomotor symptoms, such as hot flashes and night sweats, in breast cancer survivors are often worsened by chemotherapy and tamoxifen, and/or the discontinuation of hormone replacement therapy at diagnosis. This study evaluated the acceptability and effectiveness of a soy beverage containing phytoestrogens as a treatment for hot flashes in postmenopausal women with breast cancer. METHODS: A randomized, placebo-controlled, double-blind clinical trial was conducted in postmenopausal women with moderate hot flashes who were previously treated for early-stage breast cancer. Women were stratified for tamoxifen use and randomized to a soy beverage (n = 59) containing 90 mg of isoflavones or to a placebo rice beverage (n = 64). Women recorded the number and severity of hot flashes daily with a daily menopause diary for 4 weeks at baseline and for 12 weeks while consuming 500 mL of a soy or placebo beverage. RESULTS: There were no significant differences between the soy and placebo groups in the number of hot flashes or hot flash scores. However, presumably because of a strong placebo effect, both groups had significant reductions in hot flashes. Mild gastrointestinal side effects were experienced by both groups but occurred with greater frequency and severity with soy. The mean serum genistein concentration at 6 weeks was significantly higher in women who consumed soy (0.61 +/- 0.43 micromol/L) compared with placebo (0.43 +/- 0.37 micromol/L) (P =.02). Overall acceptability and compliance were high and similar in both groups. CONCLUSION: The soy beverage did not alleviate hot flashes in women with breast cancer any more than did a placebo. Future research into other compounds is recommended to identify safe and effective therapies for hot flashes in breast cancer survivors. PMID- 11896092 TI - Neoadjuvant chemotherapy in breast cancer: significantly enhanced response with docetaxel. AB - PURPOSE: To compare the efficacy of neoadjuvant (NA) docetaxel (DOC) with anthracycline-based therapy and determine the efficacy of NA DOC in patients with breast cancer initially failing to respond to anthracycline-based NA chemotherapy (CT). PATIENTS AND METHODS: Patients with large or locally advanced breast cancer received four pulses of cyclophosphamide 1,000 mg/m(2), doxorubicin 50 mg/m(2), vincristine 1.5 mg/m(2), and prednisolone 40 mg (4 x CVAP) for 5 days. Clinical tumor response was assessed. Those who responded (complete response [CR] or partial response [PR]) were randomized to receive further 4 x CVAP or 4 x DOC (100 mg/m(2)). All nonresponders received 4 x DOC. RESULTS: One hundred sixty-two patients were enrolled; 145 patients completed eight cycles of NA CT. One hundred two patients (66%) achieved a clinical response (PR or CR) after 4 x CVAP. After randomization, 50 patients received 4 x CVAP and 47 patients received 4 x DOC. In patients who received eight cycles of CT, the clinical CR (cCR) and clinical PR (cPR) (94% v 66%) and pathologic CR (pCR) (34% v 16%) response rates were higher (P =.001 and P =.04) in those who received further DOC. Intention-to-treat analysis demonstrated cCR and cPR (85% v 64%; P =.03) and pCR (31% v 15%; P =.06). Axillary lymph node examination revealed residual tumor in 33% of patients who received 8 x CVAP and 38% of patients who received further DOC. In patients who failed to respond to the initial CVAP, 4 x DOC resulted in a cCR and cPR rate of 55% and a pCR rate of 2%. Forty-four percent of these patients had residual tumor within axillary lymph nodes. CONCLUSION: NA DOC resulted in substantial improvement in responses to DOC. PMID- 11896093 TI - Elevated serum Her-2/neu level predicts decreased response to hormone therapy in metastatic breast cancer. AB - PURPOSE: To determine the effect of elevation of serum HER-2/neu on response to hormone therapy. PATIENTS AND METHODS: Seven hundred nineteen metastatic patients with estrogen receptor-positive (ER(+)), progesterone receptor-positive, or both or ER status unknown breast cancer were randomized in three independent clinical trials to receive second-line hormone therapy with either megestrol acetate or an aromatase inhibitor (fadrozole or letrozole). An automated enzyme-linked immunosorbent assay specific for the extracellular domain of the HER-2/neu (c erbB-2) oncoprotein product was used to detect serum levels. RESULTS: Two hundred nineteen patients (30%) had elevated serum HER-2/neu protein levels, using the mean + 2 SD (15 ng/mL) from the serum of healthy women as an upper limit. Response to treatment was available for 711 patients. The response rate (complete responses plus partial responses plus stable disease) to endocrine therapy was 45% in 494 patients with non-elevated and 23% in 217 patients with elevated serum HER-2/neu levels (P <.0001). Median duration of treatment response (using the time to progression [TTP] variable for patients who responded) was shorter in the group with elevated serum HER-2/neu levels (11.7 months) compared with the patient group with non-elevated levels (17.4 months). TTP, time to treatment failure, and median survival (17.2 months v 29.6 months) were also significantly shorter in the patients with elevated serum HER-2/neu levels (P <.0001). CONCLUSION: Patients with ER(+) and serum HER-2/neu-positive metastatic breast cancer are less likely to respond to hormone treatment and have a shorter duration of response than ER(+) and serum HER-2/neu-negative patients. Their survival duration is also shorter. PMID- 11896094 TI - Treatment decision making in early-stage breast cancer: should surgeons match patients' desired level of involvement? AB - PURPOSE: To describe desired and actual roles in treatment decision making among patients with early-stage breast cancer, identify how often patients' actual roles matched their desired roles, and examine whether matching of actual and desired roles was associated with type of treatment received and satisfaction. PATIENTS AND METHODS: We surveyed 1,081 women (response, 70%) diagnosed with early-stage breast cancer in Massachusetts or Minnesota about their desired and actual roles in treatment decision making with their surgeon and used logistic regression to assess whether matching of actual to desired roles was associated with type of surgery and satisfaction. RESULTS: Most patients (64%) desired a collaborative role in decision making, but only 33% reported actually having such a collaborative role when they discussed treatments with their surgeons. Overall, 49% of women reported an actual role that matched the desired role they reported, 25% had a less active role than desired, and 26% had a more active role than desired. In adjusted analyses, patients whose reported actual role matched their desired role were no more likely than others to undergo breast-conserving surgery (P >.2), but these women were more satisfied with their treatment choice (83.5% very satisfied; reference) than those whose role was less active than desired (72.9% very satisfied; P =.02) or more active than desired (72.2% very satisfied; P =.005). CONCLUSION: Only approximately half of patients reported an actual role in decision making that matched the desired role they reported. These patients were more satisfied with their treatment choice than other patients, suggesting that women with early-stage breast cancer may benefit from surgeons' efforts to identify their preferences for participation in decisions and tailor the decision making process to them. PMID- 11896095 TI - Clinical characteristics of individuals with germline mutations in BRCA1 and BRCA2: analysis of 10,000 individuals. AB - PURPOSE: To assess the characteristics that correlate best with the presence of mutations in BRCA1 and BRCA2 in individuals tested in a clinical setting. PATIENTS AND METHODS: The results of 10,000 consecutive gene sequence analyses performed to identify mutations anywhere in the BRCA1 and BRCA2 genes (7,461 analyses) or for three specific Ashkenazi Jewish founder mutations (2,539 analyses) were correlated with personal and family history of cancer, ancestry, invasive versus noninvasive breast neoplasia, and sex. RESULTS: Mutations were identified in 1,720 (17.2%) of the 10,000 individuals tested, including 968 (20%) of 4,843 women with breast cancer and 281 (34%) of 824 with ovarian cancer, and the prevalence of mutations was correlated with specific features of the personal and family histories of the individuals tested. Mutations were as prevalent in high-risk women of African (25 [19%] of 133) and other non-Ashkenazi ancestries as those of European ancestry (712 [16%] of 4379) and were significantly less prevalent in women diagnosed before 50 years of age with ductal carcinoma in situ than with invasive breast cancer (13% v 24%, P =.0007). Of the 74 mutations identified in individuals of Ashkenazi ancestry through full sequence analysis of both BRCA1 and BRCA2, 16 (21.6%) were nonfounder mutations, including seven in BRCA1 and nine in BRCA2. Twenty-one (28%) of 76 men with breast cancer carried mutations, of which more than one third occurred in BRCA1. CONCLUSION: Specific features of personal and family history can be used to assess the likelihood of identifying a mutation in BRCA1 or BRCA2 in individuals tested in a clinical setting. PMID- 11896096 TI - Women experience greater toxicity with fluorouracil-based chemotherapy for colorectal cancer. AB - PURPOSE: The toxicity profile of fluorouracil (5-FU)-based chemotherapy given on 5 consecutive days at doses of 370 to 450 mg/m(2) has been well documented. A meta-analysis of six North Central Cancer Treatment Group (NCCTG) cancer control trials involving 786 patients indicated that women treated with this type of regimen experienced more severe stomatitis and leukopenia than men. After these findings, an additional meta-analysis of the toxicity profiles on five NCCTG colorectal cancer treatment trials was undertaken. METHODS: Data for 1,093 women and 1,355 men from 12 different treatment arms were included. The primary end points were the incidence of stomatitis, leukopenia, alopecia, diarrhea, nausea, and vomiting, recorded with standard National Cancer Institute common toxicity criteria. Fisher's exact test was used to compare incidence and severity across sexes, supplemented by Forrest meta-analysis plots and logistic regression. RESULTS: The incidence of four out of six toxicities studied was significantly greater for women than men; the exceptions were severe nausea and vomiting. Overall, almost half of the women compared with a third of the men experienced severe toxicity (P <.0001). Logistic regression confirmed the univariate findings while adjusting for the effects of study, dose, body mass index, and age. The differences were consistent across treatment cycles. Response rates and survival distributions were the same for both sexes. CONCLUSION: This study confirms an earlier finding that women receiving 5-FU-based chemotherapy in a 5-day bolus schedule experience toxicity more frequently and with more severity than men. These data raise the question of whether the recommended initial dose of 5-FU based chemotherapy for women should be lower than that for men. PMID- 11896097 TI - Combined-modality treatment for resectable metastatic colorectal carcinoma to the liver: surgical resection of hepatic metastases in combination with continuous infusion of chemotherapy--an intergroup study. AB - PURPOSE: Despite technical improvements that have minimized the morbidity and mortality of hepatic surgery, the long-term outcome of resection of hepatic metastases of colorectal cancer remains poor, with the majority of patients experiencing treatment failure in the liver. Because arterial chemotherapy regimens targeted to the liver have demonstrated high response rates, an intergroup trial of adjuvant therapy for patients undergoing hepatic resection of liver metastases from colorectal cancer was initiated. PATIENTS AND METHODS: Patients with one to three potentially resectable metastases were randomized preoperatively to receive no further therapy (control arm, 56 patients) or postoperative hepatic arterial floxuridine combined with intravenous continuous infusion fluorouracil (chemotherapy arm, 53 patients). After exclusion of patients identified as ineligible for the planned treatment at the time of surgery, there were 45 control patients and 30 on the chemotherapy arm. The study was powered to evaluate improvement in time to recurrence and hepatic disease free survival, not overall survival. RESULTS: The 4-year recurrence-free rate was 25% for the control arm and 46% for the chemotherapy group (P =.04). The 4-year liver recurrence-free rate was 43% in the control group and 67% in the chemotherapy group (P =.03). The median survival of the 75 assessable patients was 49 months for the control arm and 63.7 months for the chemotherapy arm (P =.60). The median survival of all 109 patients was 47 months for the control arm compared with 34 months for the chemotherapy arm (P =.19) CONCLUSION: These data demonstrate that adjuvant intra-arterial and intravenous chemotherapy was beneficial in prolonging time to recurrence and pre-venting hepatic recurrence after hepatic resection of colorectal cancer. PMID- 11896098 TI - Volume of lymphatic metastases does not independently influence prognosis in colorectal cancer. AB - PURPOSE: To evaluate the prognostic relevance of the volume of nodal metastatic disease in colorectal cancer patients. PATIENTS AND METHODS: One hundred node positive patients with T2 or T3 carcinoma of the colon or rectum after routine histologic examination of the regional nodes were studied. The metastatic tumor was measured with an ocular micrometer, and the tumor volume was determined. RESULTS: The mean lymph node metastatic tumor volume was 5.1 +/- 4.99 mm(3) (range, 0.05 to 83,434 mm(3)). There was only a weak positive correlation with number of nodes involved with metastatic disease and tumor volume in nodes (r =.45). Median follow-up was 39 months (range, 1 to 87 months). The number of nodes was highly predictive of outcome. Individuals with one to three positive nodes had a substantially better survival than individuals with four or more positive nodes (P <.001). The volume of nodal metastatic disease correlated with outcome (P =.019). Patients dying as a result of disease had substantially greater mean metastatic nodal volume than those who were alive (3,705 v 1,783 mm(3); P =.036). However, the total metastatic nodal volume did not, independent of positive nodes or number of positive nodes, predict outcome. Individuals with micrometastatic nodal volume did not have improved survival when compared with individuals with macrometastatic nodal volume (P =.79). CONCLUSION: The number of nodes involved with metastatic tumor, rather the volume of metastatic involvement of the regional lymph nodes, predicts outcome. These results suggest that micrometastatic disease may have a similar prognosis as macrometastatic disease when the same number of lymph nodes are involved with metastatic tumor. PMID- 11896099 TI - Gemcitabine combined with oxaliplatin in advanced pancreatic adenocarcinoma: final results of a GERCOR multicenter phase II study. AB - PURPOSE: Based on preclinical in vitro synergy data, this study evaluated the activity and toxicity of a gemcitabine/oxaliplatin combination in patients with metastatic and locally advanced pancreatic adenocarcinoma. PATIENTS AND METHODS: Previously untreated metastatic and locally advanced unresectable pancreatic adenocarcinoma patients were enrolled onto this multicenter phase II study. Patients received gemcitabine 1,000 mg/m(2) as a 10-mg/m(2)/min infusion on day 1 and oxaliplatin 100 mg/m(2) as a 2-hour infusion on day 2 every 2 weeks. Patients with metastatic disease were treated until evidence of progressive disease, whereas patients with locally advanced disease received six cycles in the absence of progression, followed when appropriate by concomitant radiochemotherapy. RESULTS: Among 64 eligible patients included in eight centers, 30 had locally advanced and 34 had metastatic disease. Response rate for the 62 patients with measurable disease was 30.6% (95% confidence interval, 19.7% to 42.3%), 31.0% for locally advanced and 30.3% for metastatic patients. Among 58 assessable patients, 40% had clinical benefit. Median progression-free survival and median overall survival (OS) were 5.3 and 9.2 months, respectively, with 36% of patients alive at 1 year. Median OS for patients with metastatic disease and locally advanced disease were 8.7 and 11.5 months, respectively. With 574 treatment cycles (median per patient, nine; range, zero to 27), grade 3/4 toxicity per patient was 11% for neutropenia and thrombocytopenia, 14% for nausea or vomiting, 6.2% for diarrhea, and 11% for peripheral neuropathy, with no toxic deaths. CONCLUSION: Palliative effects, response rate, and survival observed with this well-tolerated gemcitabine/oxaliplatin combination deserve additional evaluation. A comparative study of combination therapy versus gemcitabine alone is ongoing. PMID- 11896100 TI - Randomized, open-label, phase III study of a 28-day oral regimen of eniluracil plus fluorouracil versus intravenous fluorouracil plus leucovorin as first-line therapy in patients with metastatic/advanced colorectal cancer. AB - PURPOSE: To compare the efficacy and tolerability of eniluracil (EU)/fluorouracil (5-FU) with that of 5-FU/leucovorin (LV) as first-line therapy for patients with metastatic/advanced colorectal cancer. PATIENTS AND METHODS: This multicenter, randomized, open-label, phase III study (FUMA3008) conducted in the United States and Canada compared the safety and efficacy of EU/5-FU (11.5 mg/m(2)/1.15 mg/m(2) twice daily for 28 days every 35 days) with that of intravenous 5-FU/LV (425 mg/m(2)/20 mg/m(2) once daily for 5 days every 28 days) in patients with previously untreated metastatic colorectal cancer. Overall survival (OS) was the primary end point. RESULTS: A total of 981 patients were randomized and 964 patients received treatment (485 EU/5FU, 479 5FU/LV). Survival for EU/5-FU was not statistically equivalent (but not statistically inferior) to that for 5-FU/LV (hazard ratio, 0.880; 95% confidence interval [CI], 0.75 to 1.03). Median duration of survival was 13.3 months in the EU/5-FU group and 14.5 months in the 5-FU/LV group. Median duration of progression-free survival for EU/5-FU was statistically inferior to that of the control group (20.0 weeks [95% CI, 19.1 to 20.9 weeks] v 22.7 weeks [95% CI, 18.3 to 24.6 weeks]; P =.01). Both treatments were well tolerated. Diarrhea was the most common nonhematologic toxicity in both groups; treatment-related grade 3 or 4 diarrhea occurred in 19% of patients treated with EU/5-FU and 16% of patients receiving 5-FU/LV (P =.354). Grade 3 or 4 granulocytopenia occurred in 5% of EU/5-FU patients and 47% of 5-FU/LV patients. CONCLUSION: Safety profiles of both treatments were acceptable. Although antitumor activity was observed, EU/5-FU did not meet the protocol specified statistical criteria for equivalence to 5-FU/LV in terms of OS. PMID- 11896101 TI - Simplified staging for hepatocellular carcinoma. AB - PURPOSE: The current American Joint Committee on Cancer (AJCC) staging system for hepatocellular carcinoma (HCC) fails to stratify patients adequately with respect to prognosis. PATIENTS AND METHODS: The ability of the currently proposed tumor (T) categories to effectively stratify the survival of 557 patients who underwent complete resection for HCC at four centers was examined. Independent predictors of survival were combined into a new staging system. RESULTS: Using the current AJCC T classification, patients with T1 and T2 tumors had similar 5-year survivals (P =.6). In addition, the survival of patients with multiple bilobar tumors (T4) matched that of T3 patients (P =.5). Independent predictors of death were major vascular invasion (P <.001), microvascular invasion (P =.001), severe fibrosis/cirrhosis of the host liver (P =.001), multiple tumors (P =.007), and tumor size greater than 5 cm (P =.01). Based on our results, a simplified stratification is proposed: (a) patients with a single tumor and no microvascular invasion, (b) patients with a single tumor and microvascular invasion or multiple tumors, none more than 5 cm, and (c) patients with either multiple tumors, any more than 5 cm, or tumor with major vascular invasion (P <.001). Severe fibrosis/cirrhosis had a negative impact on survival within all categories. The survival of patients with lymph node involvement matched that of patients with major vascular invasion (P =.3). CONCLUSION: The current AJCC staging system for HCC is unnecessarily complex. We propose a simplified model of stratification that is based on vascular invasion, tumor number, and tumor size and incorporates the effect of fibrosis on survival. PMID- 11896102 TI - Effect of graded testicular doses of radiotherapy in patients treated for carcinoma-in-situ in the testis. AB - PURPOSE: To determine the effect of radiotherapy in doses 14 to 20 Gy on eradication of carcinoma-in-situ (CIS) testis and on the Leydig cell function. PATIENTS AND METHODS: Forty-eight patients presented with unilateral testicular germ cell cancer and CIS of the contralateral testis. The CIS-bearing testis was treated with daily irradiation doses of 2 Gy, 5 days a week, to a cumulative dose of 20 Gy (21 patients), 18 Gy (three patients), 16 Gy (10 patients), and 14 Gy (14 patients). RESULTS: All patients treated at dose levels 20 Gy to 16 Gy achieved histologically verified complete remission without signs of recurrence of CIS after an observation period of more than 5 years. One of 14 patients treated at dose level 14 Gy had a relapse of CIS 20 months after irradiation. Leydig cell function was examined before and regularly after radiotherapy in 44 of 48 patients. The levels of testosterone were lower after radiotherapy than before. Testosterone showed a stable decrease for more than 5 years after treatment (3.6% per year) without dose dependency. The levels of luteinizing hormone and follicle-stimulating hormone were increased after radiotherapy. The need of androgen substitution therapy was similar at all dose levels. CONCLUSION: Testicular irradiation is a safe treatment at dose level 20 Gy (10 x 2 Gy). Decrease of dose to 14 Gy (7 x 2 Gy) might lead to risk of relapse of CIS. Impairment of hormone production without clinically significant dose dependency is seen in the dose range 14 to 20 Gy. PMID- 11896103 TI - Overexpression of c-met as a prognostic indicator for transitional cell carcinoma of the urinary bladder: a comparison with p53 nuclear accumulation. AB - PURPOSE: The c-met proto-oncogene encodes a receptor tyrosine kinase (Met) and has been shown to play a role in oncogenesis. Given that high titers of hepatocyte growth factor, the specific ligand for Met, are excreted in the urine and tend to reflect disease activity of bladder cancer, we performed this study to examine the clinical significance of Met in human bladder cancer. MATERIALS AND METHODS: We studied the mRNA expression and genomic alteration of c-met in five bladder cancer cell lines. Significance of Met overexpression was then compared with p53 nuclear accumulation (TP53) in primary bladder cancer (n = 142 patients). RESULTS: Expression of c-met mRNA tended to positively correlate with differentiation of cancer cell lines in the absence of point mutation. High expression of Met was found in seven cases (4.9%), low expression in 32 cases (22.5%), and negative expression in 103 cases (72.5%). Expression of Met was positively associated with histologic grade, stage classification, tumor size, and nodular tumor growth (P <.05, respectively); however, it was not related to TP53 status. Factors that predicted disease progression were tumor stage, Met status, and TP53 accumulation (P <.05, respectively). Indicators for poor long term survival were invasive cancer, multiple tumors, and Met overexpression (P =.0006,.01, and.04, respectively). CONCLUSION: The c-met proto-oncogene plays a more important role in the progression of bladder carcinogenesis than p53. Evaluation of Met expression could identify a subset of bladder cancer patients who may require a more intensive treatment strategy. PMID- 11896104 TI - Role of P53 and MDM2 in treatment response of human germ cell tumors. AB - PURPOSE: Testicular germ cell tumors (TGCTs) of adolescents and adults are very sensitive to systemic treatment. The exquisite chemosensitivity of these cancers has been attributed to a high level of wild-type P53. MATERIALS AND METHODS: To clarify the role of P53 in treatment sensitivity and resistance of TGCTs, we performed immunohistochemistry and Western blotting analysis on a series of 39 fresh-frozen primary TGCTs before therapy (unselected series). In a series of formalin-fixed paraffin-embedded TGCTs of patients with fully documented clinical course, including treatment-sensitive (n = 17) and -resistant (n = 18) tumors, P53 status was assessed by immunohistochemistry and mutation analysis. In addition, the involvement of MDM2, a P53 antagonist, was investigated by immunohistochemistry, reverse transcriptase polymerase chain reaction, and in situ hybridization. RESULTS: Immunohistochemistry demonstrated absence of staining for P53 in 36%, 41%, and 17% of the unselected, responding, and nonresponding TGCTs, respectively. Of the positive TGCTs, most tumors, ie, 49%, 41%, and 33%, showed 1% to 10% positive nuclei. This overall low level of P53 was confirmed by Western blotting. Mutation analysis revealed only one silent P53 mutation in one of the responding patients. All embryonal carcinomas were homogeneously positive for MDM2, encoded by the full length mRNA, while a heterogeneous pattern was found for the other histologic components. Amplification of MDM2 was detected in one out of 12 embryonal carcinomas. CONCLUSION: Although our results are in line with previous findings of the presence of wild-type P53 in TGCTs, they show that a high level of P53 does not relate directly to treatment sensitivity of these tumors, and inactivation of P53 is not a common event in the development of cisplatin resistance. PMID- 11896105 TI - Phase I trial of intraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015 (dl1520) given on days 1 through 5 every 3 weeks in patients with recurrent/refractory epithelial ovarian cancer. AB - PURPOSE: Resistance to chemotherapy in ovarian cancer is frequently associated with mutations in the p53 gene. The adenovirus dl1520 (ONYX-015) with the E1B 55 kd gene deleted, allowing selective replication in and lysis of p53-deficient tumor cells, has shown preclinical efficacy against p53-deficient nude mouse human ovarian carcinomatosis xenografts. PATIENTS AND METHODS: We undertook a phase I trial of intraperitoneal dl1520 in patients with recurrent ovarian cancer. Sixteen women with recurrent/refractory ovarian cancer received 35 cycles (median, two cycles) of dl1520 delivered on days 1 through 5 in four dose cohorts: 1 x 10(9) plaque forming units (pfu), 1 x 10(10) pfu, 3 x 10(10) pfu, and 1 x 10(11) pfu. RESULTS: The most common significant toxicities related to virus administration were flu-like symptoms, emesis, and abdominal pain. One patient receiving 1 x 10(10) pfu developed common toxicity criteria grade 3 abdominal pain and diarrhea, which was dose-limiting. The maximum-tolerated dose was not reached at 10(11) pfu, and at this dose level patients did not experience significant toxicity. There was no clear-cut evidence of clinical or radiologic response in any patient. Blood samples were taken for adenovirus DNA and neutralizing antibodies. Polymerase chain reaction data indicating presence of virus up to 10 days after the final (day 5) infusion of dl1520 are suggestive of continuing viral replication. CONCLUSION: This article therefore describes the first clinical experience with the intraperitoneal delivery of any replication competent/-selective virus in cancer patients. PMID- 11896106 TI - Cancer incidence in a population of Jewish women at risk of ovarian cancer. AB - PURPOSE: To evaluate the incidence and clinical characteristics of ovarian and other cancers in a cohort of women at risk of developing ovarian cancer. PATIENTS AND METHODS: The Gilda Radner Ovarian Cancer Detection Program in Los Angeles, CA, was established in 1991 to study the efficacy of screening in the early detection of ovarian cancer. We present findings from a historical cohort of 290 Jewish women who were offered BRCA testing for three common founder mutations (BRCA1 185delAG and 5382insC and BRCA2 6174delT). RESULTS: In 10 years, 17 cancers were observed (1,111 per 100,000 per year), including six breast and eight ovarian or related cancers. A high proportion of cancers of peritoneal origin was observed. The majority (86%) of women with incident breast or ovarian/peritoneal cancer carried a mutation in the BRCA1 gene. The overall cancer incidence among carriers of mutations in the BRCA1 gene was estimated to be 5,450 per 100,000 per year, corresponding to a cumulative incidence of 47.5% at 10 years. In contrast, the cumulative incidence of cancer among noncarriers was 2.5% (P < 10(-8)). After adjustment for sampling, the risks to BRCA1 mutation carriers at 10 years were estimated to be 21% for ovarian/peritoneal/tubal cancer, 16% for breast cancer, and 36% for all cancers. CONCLUSION: The excess risk of breast and ovarian cancer in Jewish women with a family history of ovarian cancer is largely attributable to mutations in BRCA1. Intensive surveillance by use of CA-125 and ultrasound does not seem to be an effective means of diagnosing early-stage ovarian cancer in this high-risk cohort. PMID- 11896107 TI - Phase III evaluation of fluoxetine for treatment of hot flashes. AB - PURPOSE: Hot flashes can be a prominent problem in women with a history of breast cancer. Given concerns regarding the use of hormonal therapies in such patients, other nonhormonal means for treating hot flashes are required. Based on anecdotal information regarding the efficacy of fluoxetine and other newer antidepressants for treating hot flashes, the present trial was developed. PATIENTS AND METHODS: This trial used a double-blinded, randomized, two-period (4 weeks per period), cross-over methodology to study the efficacy of fluoxetine (20 mg/d) for treating hot flashes in women with a history of breast cancer or a concern regarding the use of estrogen (because of breast cancer risk). Eligible patients had to have reported that they averaged at least 14 hot flashes per week; they could have received tamoxifen or raloxifene as long as they were on a stable dose. The major outcome measure was a bivariate construct representing hot flash frequency and hot flash score, analyzed by a classic sums and differences cross-over analysis. RESULTS: Eighty-one randomized women began protocol therapy. By the end of the first treatment period, hot flash scores (frequency x average severity) decreased 50% in the fluoxetine arm versus 36% in the placebo arm. Cross-over analysis demonstrated a significantly greater marked hot flash score improvement with fluoxetine than placebo (P =.02). The results were not adjusted for potential confounding influences, including age and tamoxifen use. The fluoxetine was well tolerated. CONCLUSION: This dose of fluoxetine resulted in a modest improvement in hot flashes. PMID- 11896108 TI - Time spent in hospital in the last six months of life in patients who died of cancer in Ontario. AB - PURPOSE: To describe hospital bed utilization in the final 6 months of life in patients dying of cancer in Ontario, Canada. PATIENTS AND METHODS: Hospital separation records were linked to a population-based cancer registry to identify factors associated with hospitalization in the 203,713 patients who died of cancer in Ontario between 1986 and 1998. RESULTS: Between 1986 and 1998, 5.3% of all acute care beds in Ontario were devoted to the care of cancer patients in the last 6 months of life. The mean time spent in hospital in the last 6 months of life decreased from 34.3 days in 1986 to 22.7 days in 1998. Hospitalization rates increased exponentially during the last month of life. Patients younger than 50 years of age, women, and residents of poorer communities spent significantly longer in hospital than others. Hospitalization rates differed very little among the common solid tumors, but patients with CNS malignancies, the lymphomas, and the leukemias spent significantly longer in hospital than the other groups. There was significant interregional variations in hospitalization that were not explained by differences in case mix. There was a statistically significant inverse correlation between the rate of use of palliative radiotherapy and the hospital bed use in the county in which the patient resided. CONCLUSION: The total time spent in hospital in the last 6 months of life has decreased over the last decade, but acute care hospitals continue to play a large role in the care of patients who are dying of cancer. PMID- 11896109 TI - Phase II Trial of docetaxel and cisplatin combination chemotherapy in patients with squamous cell carcinoma of the head and neck. AB - PURPOSE: To assess the antitumor activity and toxicity of docetaxel plus cisplatin chemotherapy in patients with recurrent or incurable squamous cell carcinoma of the head and neck (SCCHN). PATIENTS AND METHODS: Patients with recurrent or incurable SCCHN were eligible if they were chemotherapy naive or if they had received one prior regimen as neoadjuvant or concurrent treatment with radiation. Patients who had received chemotherapy for recurrence or prior taxanes were ineligible. Patients received docetaxel 75 mg/m(2) and cisplatin 75 mg/m(2) on day 1; cycles were repeated every 21 days. RESULTS: Toxic effects and length of survival were assessable in 36 patients and tumor response was assessable in 32, for whom the overall response rate was 40% (13 of 32) (6% complete response and 34% partial response). Median time to response was 5 weeks, and median duration was 4.9 months. In the intent to treat population (n = 36), median time to disease progression was 4 months. Median survival (n = 36) was 9.6 months, and the 12-month survival rate was 27%. Grade 4 neutropenia was observed in 71% of patients. Two patients (6%) experienced serious fever during grade 4 neutropenia (without documented infection) that required intravenous antibiotics, and an additional four patients had grade 3 infection. Other severe (grades 3 and 4) toxic effects were asthenia (25%), nausea (11%), fever (8%), vomiting (8%), severe hypersensitivity reactions (8%), and diarrhea (8%). Severe stomatitis (grade 3) occurred in only one patient. CONCLUSION: Docetaxel plus cisplatin is an effective regimen with an acceptable safety profile for palliation of recurrent SCCHN. Relative to the standard regimen of cisplatin/fluorouracil, this regimen may offer higher tumor response and survival rates with short outpatient administration and a lower incidence of severe mucosal toxicity. PMID- 11896110 TI - Cisplatin, dacarbazine with or without subcutaneous interleukin-2, and interferon alpha-2b in advanced melanoma outpatients: results from an Italian multicenter phase III randomized clinical trial. AB - PURPOSE: Phase II and III studies have shown that the addition of interleukin-2 (IL-2) and interferon alpha-2b (IFN alpha-2b) in multiagent chemotherapy (CT) for advanced melanoma increases overall response (OR), albeit without clear evidence of an improvement in overall survival (OS). Treatment with high-dose IL-2 can cause severe toxicity and is normally administered in an inpatient setting. We conducted a multicenter prospective randomized clinical trial in outpatients with metastatic melanoma to compare CT with biochemotherapy (bioCT) using immunomodulant doses of IL-2 and IFN alpha-2b. PATIENTS AND METHODS: One hundred seventy-six eligible patients with advanced melanoma were randomized to receive CT (cisplatin and dacarbazine with or without carmustine every 21 days) or bioCT comprising the same CT regimen followed by low-dose subcutaneous IL-2 for 8 days and IFN alpha-2b three times a week, both for six cycles. RESULTS: At a median follow-up of 18 (CT) and 16 (bioCT) months, median OS was 9.5 versus 11.0 months (P =.51), respectively. In the 89 CT-arm patients, 18 ORs (20.2%) (three complete responders [CRs] and 15 partial responders [PRs]) were observed according to World Health Organization criteria. In the 87 bioCT-arm patients, 22 ORs (25.3%) (three CRs and 19 PRs) (P =.70) were recorded. Treatment-related toxicity was fairly similar in both arms. CONCLUSION: The addition of low-dose immunotherapy did not produce a statistically significant advantage in OS, time to progression, or OR. However, the 11-month median OS in the bioCT arm does not differ greatly from the best results with high-dose IL-2-containing regimens reported in the literature. Furthermore, our treatment schedule was carried out on outpatients and had an acceptable level of toxicity. PMID- 11896111 TI - Predictors of smoking initiation and cessation among childhood cancer survivors: a report from the childhood cancer survivor study. AB - PURPOSE: To examine the determinants of smoking behavior among participants in the Childhood Cancer Survivors Study (CCSS). METHODS: This retrospective cohort survey study was conducted among 9,709 childhood cancer survivors. Main outcomes included smoking initiation and cessation. RESULTS: Twenty-eight percent of patients reported ever smoking and 17% reported being current smokers. Standardized to United States population rates, the observed to expected (O/E) ratios and corresponding 95% confidence limits (95% CL) of cigarette smoking were 0.72 (95% CL, 0.69, 0.75) among all survivors and 0.71 (95% CL, 0.68 to 0.74) and 0.81 (95% CL, 0.70, 0.93) among whites and nonwhites, respectively. Significantly lower O/E ratios were present among both males (O/E, 0.73) and females (O/E, 0.70). Factors independently associated with a statistically significant relative risk of smoking initiation included older age at cancer diagnosis, lower household income, less education, not having had pulmonary-related cancer treatment, and not having had brain radiation. Blacks were less likely to start smoking. Survivors who smoked were significantly more likely to quit (O/E, 1.22; 95% CL, 1.15, 1.30). Among ever-smokers, factors associated with the likelihood of being a current smoker included age less than 13 years at smoking initiation, less education, and having had brain radiation; those age less than 3 years at cancer diagnosis were significantly more likely to be ex-smokers. CONCLUSIONS: Although survivors in the CCSS cohort seem to be smoking at rates below the general population, interventions are needed to prevent smoking initiation and promote cessation in this distinct population. PMID- 11896112 TI - Protracted intermittent schedule of topotecan in children with refractory acute leukemia: a pediatric oncology group study. AB - PURPOSE: To determine dose-limiting toxicity (DLT) and maximum-tolerated dose (MTD) of a protracted, intermittent schedule of daily 30-minute infusions of topotecan (TPT) for up to 12 consecutive days, every 3 weeks, in children with refractory leukemia. PATIENTS AND METHODS: Forty-nine children were enrolled onto this phase I trial (24 with acute nonlymphoblastic leukemia [ANLL] and 25 with acute lymphoblastic leukemia [ALL]). TPT dosage was escalated from 2.0 to 5.2 mg/m(2)/d for 5 days and 2.4 mg/m(2)/d from 7 days to the same dose for 9 and 12 days in cohorts of three to six patients when no DLT was identified. TPT pharmacokinetics were studied in 33 children once or twice (first and last doses in patients who received TPT for > 7 days). RESULTS: Seventy assessable courses of TPT were administered to 49 children who had refractory leukemia. DLTs were typhlitis, diarrhea, and mucositis, and the MTD was 2.4 mg/m(2)/d for 9 days in this group of heavily pretreated children. In 33 patients, the median TPT lactone clearance after the first dose was 19.2 L/h/m(2) (range, 9.4 to 45.9 L/h/m(2)) and did not change during the course. There were significant responses (one complete response [CR] and four partial responses [PR] in patients with ANLL and one CR and two PRs in patients with ALL), and all but one were at dosages of TPT given for at least 9 days. CONCLUSION: The MTD was 2.4 mg/m(2)/d for 9 days. Further testing is warranted of TPT's schedule dependence in children with leukemia. PMID- 11896113 TI - Prognostic factors for malignant transformation in monoclonal gammopathy of undetermined significance and smoldering multiple myeloma. AB - PURPOSE: To evaluate the natural history of monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM), identify early predictors of evolution, and assess whether associated conditions correlate with disease progression. PATIENTS AND METHODS: A total of 1,231 consecutive patients with either MGUS (n = 1,104) or SMM (n = 127) diagnosed from July 1975 to March 1998 were included in the study. Cumulative survival probability and cumulative probability of transformation into lymphoproliferative disease were calculated by means of the Kaplan-Meier estimator. Univariate and multivariate Cox models were used to identify possible predictors of malignant evolution. RESULTS: Cumulative transformation probability at 10 and 15 years was 14% and 30%, respectively. At a median follow-up of 65 months (range, 12 to 239 months), 64 MGUS cases (5.8%) evolved to multiple myeloma (MM) (n = 43), extramedullary plasmacytoma (n = 1), primary amyloidosis (n = 1), Waldenstrom's macroglobulinemia (n = 12), non Hodgkin's lymphoma (n = 6), and B-chronic lymphocytic leukemia (n = 1). At a median follow-up of 72 months (range, 12 to 247 months), 25 SMMs (19.7%) evolved to overt MM. A lower evolution risk was observed in MGUS than in SMM (P <.0001). Greater than 5% marrow plasmacytosis, detectable Bence Jones proteinuria, polyclonal serum immunoglobulin reduction, and high erythrocyte sedimentation rate (ESR) were independent factors influencing MGUS transformation. SMM progression correlated with greater than 10% marrow plasma cells, detectable Bence Jones proteinuria, and immunoglobulin (Ig) A isotype. Neither concomitant diseases nor immunosuppression correlated with progression. CONCLUSION: Careful evaluation of marrow plasmacytosis, urinary paraprotein, background immunoglobulins, ESR, and paraprotein isotype might help identify at presentation patients with benign monoclonal gammopathies requiring stricter monitoring. PMID- 11896114 TI - Survival and failure patterns of high-grade gliomas after three-dimensional conformal radiotherapy. AB - PURPOSE: The goal of three-dimensional (3-D) conformal radiation is to increase the dose delivered to tumor while minimizing dose to surrounding normal brain. Previously it has been shown that even escalated doses of 70 to 80 Gy have failure patterns that are predominantly local. This article describes the failure patterns and survival seen with high-grade gliomas given 90 Gy using a 3-D conformal intensity-modulated radiation technique. PATIENTS AND METHODS: From April 1996 to April 1999, 34 patients with supratentorial high-grade gliomas were treated to 90 Gy. For those that recurred, failure patterns were defined in terms of percentage of recurrent tumor located within the high-dose region. Recurrences with more than 95% of their volume within the high-dose region were considered central; those with 80% to 95%, 20% to 80%, and less than 20% were considered in field, marginal, and distant, respectively. RESULTS: The median age was 55 years, and median follow-up was 11.7 months. At time of analysis, 23 (67.6%) of 34 patients had developed radiographic evidence of recurrence. The patterns of failure were 18 (78%) of 23 central, three (13%) of 23 in-field, two (9%) of 23 marginal, and zero (0%) of 23 distant. The median survival was 11.7 months, with 1-year survival of 47.1% and 2-year survival of 12.9%. No significant treatment toxicities were observed. CONCLUSION: Despite dose escalation to 90 Gy, the predominant failure pattern in high-grade gliomas remains local. This suggests that close margins used in highly conformal treatments do not increase the risk of marginal or distant recurrences. Our results indicate that intensification of local radiotherapy with dose escalation is feasible and deserves further evaluation for high-grade gliomas. PMID- 11896115 TI - Adjuvant radiation for stage II-B soft tissue sarcoma of the extremity. AB - PURPOSE: Adjuvant radiation therapy (RT) has been shown to improve local control in patients with high-grade soft tissue sarcoma (STS) of the extremity. This study sought to define the optimal management in patients with stage II-B (high grade, size < or = 5 cm) tumors. PATIENTS AND METHODS: Between July 1982 and December 1998, 204 adult patients with primary stage II-B STS underwent limb sparing surgery with negative microscopic margins. Eighty-eight patients (43%) received RT; 116 (57%) did not. The RT and no-RT groups were balanced with regard to age, site (upper v lower extremity), whether patients had prior unplanned excision, and location (central, i.e., shoulder/groin v non-central). The RT group had more deep tumors (P =.03). Adjuvant RT was delivered with brachytherapy in 60% and external-beam radiation in 40% of patients. RESULTS: With a median follow-up of 67 months, the 5-year local control, distant relapse-free survival, and disease-specific survival rates were 82%, 80%, and 88%, respectively. There was no significant difference in local control between the RT and no-RT groups (84% v 80%, respectively, P =.3). Tumor depth, site, and prior unplanned excision did not correlate with local control. The only independent predictors of poor local control were central tumor location (relative risk [RR] = 3; 95% confidence interval [CI], 2 to 7; P =.005) and age more than 50 years (RR = 6; 95% CI, 2 to 13; P =.001). CONCLUSION: In this retrospective study, adjuvant RT did not significantly improve local control in patients with stage II-B STS of the extremity. The outcome of patients with central tumor location was poor, and efforts to identify the optimal local treatment approach for such patients are warranted. PMID- 11896116 TI - Gemcitabine, carboplatin, and paclitaxel for patients with carcinoma of unknown primary site: a Minnie Pearl Cancer Research Network study. AB - PURPOSE: To evaluate the efficacy and toxicity of the novel chemotherapy combination that includes gemcitabine, carboplatin, and paclitaxel in the treatment of patients with carcinoma of unknown primary site. PATIENTS AND METHODS: One hundred twenty patients were treated with the following regimen, administered every 21 days for a planned four courses: gemcitabine 1,000 mg/m(2) intravenously (i.v.) on days 1 and 8, carboplatin at an estimated area under the concentration-time curve of 5 mg min/mL i.v. on day 1, and paclitaxel 200 mg/m(2) i.v. on day 1. After four courses, stable and responding patients were given weekly paclitaxel 70 mg/m(2) i.v. for 6 weeks for three 8-week courses. All patients had relatively poor prognostic features. Sixty-three patients had well differentiated adenocarcinoma, 56 patients had poorly differentiated carcinoma, and 104 patients had performance status of 0 or 1. RESULTS: Twenty-eight (25%) of 113 assessable patients (95% confidence interval, 22% to 30%) had major objective responses to treatment. Response rates were similar in the two major histologic types. Response rate did not seem to be improved by continued therapy with weekly paclitaxel. The median progression-free survival time was 6 months. Median survival for the entire group was 9 months, and the actuarial survival at 1 and 2 years was 42% and 23%, respectively. CONCLUSION: Combination chemotherapy with gemcitabine, carboplatin, and paclitaxel followed by weekly paclitaxel is an active and tolerable treatment for patients with carcinoma of unknown primary site. The survival seen in this poor-prognosis group of patients in this multicenter community-based trial is notable and similar to other taxane-based regimens for these patients. Study of additional combinations or sequences of newer drugs, as well as the exploration of targeted biologic agents for patients with an identified target in their tumors, is warranted. PMID- 11896117 TI - Dose-finding and pharmacokinetic study of cisplatin, gemcitabine, and SU5416 in patients with solid tumors. AB - PURPOSE: To investigate the feasibility and pharmacokinetics of the combination cisplatin, gemcitabine, and SU5416. PATIENTS AND METHODS: Patients received cisplatin 80 mg/m(2) on day 1, gemcitabine 1,250 mg/m(2) on days 1 and 8, repeated every 3 weeks, and SU5416 (85 and 145 mg/m(2)) intravenously twice weekly. Pharmacokinetics of all three agents, side effects, and antitumor response were investigated in patients with solid tumors amenable to therapy with cisplatin/gemcitabine. RESULTS: In the first cohort of three patients entered at the 85 mg/m(2) dose, no dose-limiting toxicities were observed. In the next cohort (145 mg/m(2)), three patients developed a thromboembolic event. After entry was restricted to patients with low thromboembolic risk, three additional patients enrolled at 145 mg/m(2) developed a thromboembolic event. The dose was then reduced to 85 mg/m(2) in all patients still on the study, and three additional patients were entered on this dose level. In 19 treated patients, eight patients developed nine thromboembolic events (three transient ischemic attacks, two cerebrovascular accidents, and four deep venous thromboses). The most common toxicities observed were those previously reported for SU5416 alone (headache and phlebitis) and for this chemotherapy regimen (nausea, thrombocytopenia, and leucopenia). No significant pharmacologic interaction among the three drugs was observed. Response rates were similar to those expected in the patient population selected for this study. Analysis of variables of the coagulation cascade and of vessel wall activation was performed in three patients and showed significant increases in thrombin generation and endothelial cell perturbation in a treatment cycle-dependent manner. CONCLUSION: The incidence of thromboembolic events, possibly related to the particular regimen tested in this study, discourages further investigation of this regimen. PMID- 11896118 TI - Hepatic drug targeting: phase I evaluation of polymer-bound doxorubicin. AB - PURPOSE: Preclinical studies have shown good anticancer activity following targeting of a polymer bearing doxorubicin with galactosamine (PK2) to the liver. The present phase I study was devised to determine the toxicity, pharmacokinetic profile, and targeting capability of PK2. PATIENTS AND METHODS: Doxorubicin was linked via a lysosomally degradable tetrapeptide sequence to N-(2 hydroxypropyl)methacrylamide copolymers bearing galactosamine. Targeting, toxicity, and efficacy were evaluated in 31 patients with primary (n = 25) or metastatic (n = 6) liver cancer. Body distribution of the radiolabelled polymer conjugate was assessed using gamma-camera imaging and single-photon emission computed tomography. RESULTS: The polymer was administered by intravenous (i.v.) infusion over 1 hour, repeated every 3 weeks. Dose escalation proceeded from 20 to 160 mg/m(2) (doxorubicin equivalents), the maximum-tolerated dose, which was associated with severe fatigue, grade 4 neutropenia, and grade 3 mucositis. Twenty-four hours after administration, 16.9% +/- 3.9% of the administered dose of doxorubicin targeted to the liver and 3.3% +/- 5.6% of dose was delivered to tumor. Doxorubicin-polymer conjugate without galactosamine showed no targeting. Three hepatoma patients showed partial responses, with one in continuing partial remission 47 months after therapy. CONCLUSION: The recommended PK2 dose is 120 mg/m(2), administered every 3 weeks by IV infusion. Liver-specific doxorubicin delivery is achievable using galactosamine-modified polymers, and targeting is also seen in primary hepatocellular tumors. PMID- 11896119 TI - Doxorubicin administration by continuous infusion is not cardioprotective: the Dana-Farber 91-01 Acute Lymphoblastic Leukemia protocol. AB - PURPOSE: Acute doxorubicin-induced cardiotoxicity can be prevented in adults by continuous infusion of the drug, but mechanisms of cardiotoxicity are different in children. We compared cardiac outcomes in children receiving bolus or continuous infusion of doxorubicin. PATIENTS AND METHODS: In a randomized study, children with high-risk acute lymphoblastic leukemia received doxorubicin 360 mg/m(2) in 30-mg/m(2) doses every 3 weeks either by bolus (within 1 hour, n = 57) or by continuous infusion (over 48 hours, n = 64). Echocardiograms obtained before doxorubicin and at longest follow-up times were centrally remeasured, and z scores of cardiac measurements were calculated based on a healthy population. RESULTS: The groups were similar in age, sex distribution, doxorubicin dose, and duration of follow-up. Before treatment, measures of left ventricular (LV) structure and function did not reveal dilated cardiomyopathy and were not statistically different between bolus and continuous-infusion groups. The follow up echocardiograms demonstrated no significant difference between the two groups for any cardiac characteristic, but both groups showed significant abnormalities of LV structure and function compared with normal and with baseline. For example, the mean LV fractional shortening fell by approximately two SD in both groups between the two echocardiograms. LV contractility was depressed in both groups (for bolus patients, median z score = -0.70 SD, P =.006; for continuous-infusion patients, median z score = -0.765, P =.005). Dilated cardiomyopathy and inadequate LV hypertrophy were noted in both groups. Clinical cardiac manifestations and event-free survival did not differ. CONCLUSION: Continuous doxorubicin infusion over 48 hours for childhood leukemia did not offer a cardioprotective advantage over bolus infusion. Both regimens were associated with progressive subclinical cardiotoxicity. Other cardioprotective strategies should be explored. PMID- 11896121 TI - Inhibition of KIT tyrosine kinase activity: a novel molecular approach to the treatment of KIT-positive malignancies. AB - PURPOSE: Activation of the KIT tyrosine kinase by somatic mutation has been documented in a number of human malignancies, including gastrointestinal stromal tumor (GIST), seminoma, acute myelogenous leukemia (AML), and mastocytosis. In addition, paracrine or autocrine activation of this kinase has been postulated in numerous other malignancies, including small-cell lung cancer and ovarian cancer. In this review, we discuss the rationale for and development of KIT tyrosine kinase inhibitors for the treatment of human malignancies. MATERIALS AND METHODS: Studies were identified through a MEDLINE search, review of bibliographies of relevant articles, and review of abstracts from national meetings. RESULTS: Four tyrosine kinase inhibitors that have activity against KIT are currently being used in clinical trials, and one, STI571, has recently been approved by the United States Food and Drug Administration for treating patients with chronic myelogenous leukemia. The role of KIT inhibitors in treating KIT-positive malignancies is reviewed. CONCLUSION: Targeted therapy to inhibit the kinase activity of KIT is a rational approach to the treatment of KIT-positive malignancies. Two key factors are the potency of a given inhibitor and the relative contribution of KIT activation to the growth of the tumor. Given our current understanding of KIT activity in human malignancy, the best candidate diseases for treatment with KIT inhibitors are GIST, mastocytosis, seminoma and possibly some cases of AML. Additionally, KIT inhibitors may play an adjunctive role in diseases such as small-cell lung cancer, in which KIT activation is secondary to ligand binding rather than an acquired mutation. PMID- 11896120 TI - Comparative pharmacokinetic study of continuous venous infusion fluorouracil and oral fluorouracil with eniluracil in patients with advanced solid tumors. AB - PURPOSE: To compare the pharmacokinetics of continuous venous infusion (CVI) fluorouracil (5-FU) with that of oral eniluracil/5-FU and to describe toxicities and clinical activity of prolonged oral administration of eniluracil/5-FU. PATIENTS AND METHODS: A randomized, open-label, cross-over study compared CVI 5 FU to an oral 5-FU/eniluracil combination. Seventeen patients (arm A) were randomly assigned to receive eniluracil/5-FU combination tablets (10:1 mg/m(2) BID for 7 days) during the first study period, followed by 5-FU (300 mg/m(2) CVI for 7 days) during period 2, with a 14-day washout between periods. Sixteen patients (arm B) received treatment in the opposite sequence. In period 3, all patients received eniluracil/5-FU tablets BID for 28 days. Plasma levels of 5-FU during CVI and oral administration were analyzed in periods 1 and 2. Dihydropyrimidine dehydrogenase (DPD) activity was determined by measuring plasma uracil, urinary alpha-fluoro-beta-alanine, and peripheral-blood mononuclear cell (PBMC) DPD activity. RESULTS: There were no grade 3 or 4 toxicities in either arm. Partial responses were observed in three patients. Another three patients had stable disease for > or = 3 months. Eniluracil and 5-FU pharmacokinetics were similar to those observed in previous studies and were unaffected by administration sequence. The mean +/- SD steady-state plasma concentration (C(P)) and area under the curve (AUC)(144-168h) for CVI 5-FU (104 +/- 45 ng/mL and 2,350 +/- 826 ng x h/mL, respectively) were three-fold greater than those for oral 5-FU (38.1 +/- 7.7 ng/mL and 722 +/- 182 ng x h/mL, respectively [P <.00001]). Individual 5-FU concentrations during CVI were highly variable, whereas those after eniluracil/5-FU were very reproducible. DPD activity in PBMCs before each study period was normal. CONCLUSION: Both CVI 5-FU and oral eniluracil/5-FU were well tolerated, with moderate activity in these heavily pretreated patients. However, 5-FU steady-state C(P) and AUCs achieved with oral eniluracil/5-FU were significantly less than with CVI 5-FU. PMID- 11896122 TI - Quality of life at the end of life: how is the solution far away? PMID- 11896123 TI - Multimodal molecular screening is required to improve the sensitivity of MLH1 and MSH2 mutation analysis. PMID- 11896124 TI - Use of trade names of drugs and the scientific content of medical congresses. PMID- 11896125 TI - Following a trial that stopped early: what next for adjuvant hepatic intra arterial iodine-131 lipiodol in resectable hepatocellular carcinoma? PMID- 11896128 TI - Violence against women: a global public health issue! PMID- 11896129 TI - Psychosocial risk factors at the workplace: is there enough evidence to establish reference values? PMID- 11896130 TI - Can lay-led walking programmes increase physical activity in middle aged adults? A randomised controlled trial. AB - STUDY OBJECTIVE: To compare health walks, a community based lay-led walking scheme versus advice only on physical activity and cardiovascular health status in middle aged adults. DESIGN: Randomised controlled trial with one year follow up. Physical activity was measured by questionnaire. Other measures included attitudes to exercise, body mass index, cholesterol, aerobic capacity, and blood pressure. SETTING: Primary care and community. PARTICIPANTS: 260 men and women aged 40-70 years, taking less than 120 minutes of moderate intensity activity per week. MAIN RESULTS: Seventy three per cent of people completed the trial. Of these, the proportion increasing their activity above 120 minutes of moderate intensity activity per week was 22.6% in the advice only and 35.7% in the health walks group at 12 months (between group difference =13% (95% CI 0.003% to 25.9%) p=0.05). Intention to treat analysis, using the last known value for missing cases, demonstrated smaller differences between the groups (between group difference =6% (95% CI -5% to 16.4%)) with the trend in favour of health walks. There were improvements in the total time spent and number of occasions of moderate intensity activity, and aerobic capacity, but no statistically significant differences between the groups. Other cardiovascular risk factors remained unchanged. CONCLUSIONS: There were no significant between group differences in self reported physical activity at 12 month follow up when the analysis was by intention to treat. In people who completed the trial, health walks was more effective than giving advice only in increasing moderate intensity activity above 120 minutes per week. PMID- 11896131 TI - Health inequalities by education and age in four Nordic countries, 1986 and 1994. AB - STUDY OBJECTIVE: To compare the age pattern of educational health inequalities in four Nordic countries in the mid-1980s and the mid-1990s. DESIGN: Cross sectional interview surveys at two points of time. SETTING: Data on self reported limiting longstanding illness, and perceived health were collected from Denmark, Finland, Norway, and Sweden in 1986/87 and in 1994/95. PARTICIPANTS: Representative samples of the non-institutionalised population at 15 years or older. Analyses were restricted to respondents aged between 25 and 75 (n= 23 325 men and 24 184 women). Response rates varied from 73% to 87%. MAIN RESULTS: The age adjusted prevalence of limiting longstanding illness in Finland was 10% higher in men and 6% higher in women than in other Nordic countries in 1986/87 but the gap narrowed by 1994/95. Educational health inequalities were largest in Norway. In 1986/87 the odds ratio (OR) for limiting longstanding illness was 11.25 (95% CI 8.66 to 14.62) among men and 8.23 (95% CI 6.60 to 10.27) among women in the oldest age group (65-74 years old) in Finland when the youngest age group (25-34 years old) was used as the reference category (OR=1.00). The age pattern in Finland was steeper than in Sweden (OR=5.02, 95% CI 3.97 to 6.34 in men and 5.29, 95% CI 4.18 to 6.71 in women) or Norway (OR=6.32, 95% CI 4.06 to 9.84 and 5.45, 95% CI 3.81 to 7.82, respectively). In 1994/95 relative health improved in the oldest age group in Finland (OR=5.80, 95% CI 4.33 to 7.78 in men and 5.94, 95% CI 4.52 to 7.79 in women) and in Norway (OR=4.55, 95% CI 3.01 to 6.88 and 3.96, 95% CI 2.70 to 5.81, respectively) but remained stable in Sweden. The study compared health differences by age in different educational categories and found that in Finland in 1986/87 the health in the oldest age group was poorer for secondary (OR=10.59, 95% CI 5.96 to 18.82) or basic educated (OR=9.76, 95% CI 6.66 to 14.30) men than for men with higher education (OR=5.15, 95% CI 2.59 to 10.22). The difference was not found among women or in other Nordic countries and it diminished among men in Finland in 1994/95. The results of perceived health were broadly similar to the above results of limiting longstanding illness. CONCLUSION: The results suggest that compared with other Nordic countries the comparatively poorer health in Finland is partly attributable to a cohort effect. This may be associated with the lower standard of living in Finland that lasted until the mid-1950s. The cohort effect is also likely to contribute to educational health inequalities among older Finnish men. The results suggest that not only current social policies but also past economic circumstances are likely to affect the overall health status as well as health inequalities. PMID- 11896132 TI - Liberal fortification of foods: the risks. A study relating to Finland. AB - STUDY OBJECTIVE: The free circulation of goods in the European Union (EU) has increased the number of fortified foods available to consumers in Finland. Fortification of foodstuffs with calcium is currently widespread. To assist decision making relating to fortification, the object of this study was to determine whether there might be a risk of excessive levels of calcium. The study was done using the dietary data of adult population by sex and calcium intake deciles, taking account of intakes of calcium from normal diets and from fortified foods. STUDY DESIGN: Calcium intakes for deciles were calculated from dietary data of the population involved in the 1992 FINDIET survey. Calcium fortification was evaluated by considering foodstuffs to which calcium has already been added or which producers might wish to fortify and market in Finland. Situations in which a fortified product replaced its unfortified equivalent were envisaged. Daily calcium intakes (mg) from particular foodstuffs were calculated based on known amounts of calcium (mg) per 100 g of each foodstuff and amounts of each product consumed per day. Total calcium intakes of people in different deciles were calculated by adding amounts of calcium contributed by each fortified product to normal dietary intakes. Calcium intakes were also calculated for consumption by persons of all foodstuffs that are or intended to be fortified with calcium. STUDY PARTICIPANTS: Adults aged 25 to 64 representing populations in four regions of Finland, who recorded food consumption over three day periods in spring 1992. STUDY SETTING: North Karelia, Kuopio Province, Turku-Loimaa, and the capital area Helsinki-Vantaa, Finland. MAIN STUDY RESULTS: The results indicate that the highest 10% of Finnish adult men receive on average 2315 mg calcium per day from diets that do not include fortified foodstuffs. These people whose energy consumption was high consumed substantial amounts of dairy products. Consumption of fortified foods would increase the calcium intake further to levels exceeding the tolerable upper intake level (2500 mg/d). CONCLUSIONS: Liberal addition of calcium to various groups of foodstuffs could increase the calcium intake in the highest decile to levels with potentially untoward health effects. In assessing benefits and possible disadvantages of fortification of foodstuffs, the dietary habits of the population concerned, probable consumption of fortified products and nutritional intakes, especially at extremes, need to be taken into account, by total risk assessment. PMID- 11896133 TI - Sexual assault among North Carolina women: prevalence and health risk factors. AB - STUDY OBJECTIVE: Sexual assault is traumatic at the time it occurs, but it also may have longlasting negative effects on physical health. Much of the research linking specific health problems to sexual assault victimization has used samples from special populations. The goals of this study are to estimate the prevalence of sexual assault in a representative sample of women in North Carolina and examine sexual assault in relation to specific health risk factors for leading causes of morbidity and mortality in women. DESIGN: The North Carolina Behavioral Risk Factor Surveillance System (BRFSS) is a household telephone survey of non institutionalised adults, 18 years of age and older, conducted by random digit dialling. SETTING: This investigation focuses on the study participants in the 1997 survey. PARTICIPANTS: The sample includes 2109 women who responded to the sexual assault questions in the 1997 North Carolina BRFSS interview. MAIN RESULTS: The lifetime prevalence of sexual assault was 19% (95% CI 17% to 20%), of which 73% of victims experienced or were threatened with forced sexual intercourse. Sexual assault victims, particularly victims of forced intercourse or the threat thereof, were more likely to perceive their general health as being fair or poor (OR=2.3, 95% CI 1.5 to 3.4) and were more likely to have suffered poor physical and mental health in the past month (poor physical health, OR=2.1, 95% CI 1.6 to 2.8; poor mental health, OR= 2.6, 95% CI 1.9 to 3.5). After controlling for sociodemographic factors and health care coverage, victims of forced intercourse or the threat thereof were more likely to smoke cigarettes (OR=2.0, 95% CI 1.4 to 2.8), to have hypertension (OR=1.5, 95% CI 1.1 to 2.2), to have high cholesterol (OR=1.7, 95% CI 1.2 to 2.5), and to be obese (OR=1.7, 95% CI 1.3 to 2.4). CONCLUSIONS: This study shows associations between sexual victimization and health risk factors in a non-clinical statewide population of women. Future research should determine whether clinically screening for and identifying a history of sexual victimization among women seen in a variety of health care settings leads to the initiation of effective interventions that help women successfully cope with these violent experiences. There is also a need for further research to investigate the temporal sequence of assaults and subsequent health outcomes by assessing physical health status before and after victimization. PMID- 11896134 TI - Employee control over working times: associations with subjective health and sickness absences. AB - STUDY OBJECTIVE: To investigate the impact of employees' worktime control on health, taking into account other aspects of job control. DESIGN: Analysis of questionnaire data in 1997 and register data on sickness absence during 1996 1998. SETTING: Eight towns in Finland. PARTICIPANTS: 6442 municipal employees (1490 men and 4952 women) representing the staff of the towns studied. Follow up was 17 706 person years. MAIN RESULTS: In women, poor health and psychological distress were more prevalent among those in the lowest quartile of worktime control than those in the highest (after adjustment for potential confounders including other aspects of job control, odds ratios and their 95% confidence intervals for poor health and psychological distress were 1.8 (1.5 to 2.3) and 1.6 (1.3 to 2.0), respectively). Correspondingly, the adjusted sickness absence rate was 1.2 (1.1 to 1.2) times higher in women with low worktime control than in women with high worktime control. In men, no significant associations between worktime control and health were found. These results, obtained from the total sample, were replicable within a homogeneous occupational group comprising women and men. CONCLUSIONS: Exploration of specific aspects of job control provides new information about potentially reversible causes of health problems in a working population. Worktime control is an independent predictor of health in women but not in men. Dissimilarities in the distribution of occupations between men and women are not a probable explanation for this difference. PMID- 11896135 TI - Higher blood pressure among Inuit migrants in Denmark than among the Inuit in Greenland. AB - STUDY OBJECTIVE: Previous studies of blood pressure among the Inuit have given inconsistent results and studies comparing Inuit migrants with those living in traditional Inuit areas are absent. The purpose of the study was to compare the blood pressure of the Inuit in Greenland with that of Inuit migrants in Denmark. DESIGN: Questionnaire, interview, and clinical examination in a cross sectional random population sample. SETTING: A population based survey among Inuit in Greenland and Inuit migrants in Denmark. PARTICIPANTS: 2046 Inuit aged >/or =18, 61% of the sample. MAIN RESULTS: Age and gender adjusted blood pressures were 117/72 mm Hg in Greenland and 127/81 mm Hg among the migrants (p<0.001). In both populations, blood pressure increased with age and body mass index, and was higher among men and non-smokers. In Greenland, blood pressure increased with the level of school education. The associations with Inuit heritage, alcohol, diet, and physical activity were not significant. The difference between the two populations persisted after adjusting for age, gender, body mass index, education, and smoking. Among those who had completed high school, there was no difference between the systolic blood pressure of the two populations while the difference for diastolic blood pressure was much less than for those with less education. CONCLUSIONS: Blood pressure was lower among the Inuit in Greenland than among the Inuit migrants in Denmark but the difference was absent (systolic pressure) or reduced (diastolic pressure) among the better educated. The results suggest that the blood pressure of the Inuit, especially Inuit men, may be responsive to factors related to the modern Western way of life. PMID- 11896136 TI - Trends in head injury mortality among 0-14 year olds in Scotland (1986-95). AB - STUDY OBJECTIVE: To examine the trends in childhood head injury mortality in Scotland between 1986 and 1995. DESIGN: Analysis of routine mortality data from the registrar general for Scotland. SETTING: Scotland, UK. SUBJECTS: Children aged 0-14 years. MAIN RESULTS: A total of 290 children in Scotland died as a result of a head injury between 1986 and 1995. While there was a significant decline in the head injury mortality rate, head injury as a proportion of all injury fatalities remained relatively stable. Boys, and children residing in relatively less affluent areas had the highest head injury mortality rates. Although both these groups experienced a significant decline over the study period, the mortality differences between children in deprivation categories 1-2 and 6-7 persisted among 0-9 year olds, and increased in the 10-14 years age group. Pedestrian accidents were the leading cause of mortality. CONCLUSIONS: Children residing in less affluent areas seem to be at relatively greater risk of sustaining a fatal head injury than their more affluent counterparts. While the differences between the most and least affluent have decreased overall, they have widened among 10-14 year olds. The decline in head injury mortality as a result of pedestrian accidents may be partly attributable to injury prevention measures. PMID- 11896137 TI - Use of acute hospital beds does not increase as the population ages: results from a seven year cohort study in Germany. AB - OBJECTIVES: (1) To compare the number of hospital days used by survivors with those by persons in their last, second last, and third last year of life in relation to age; (2) to analyse lifelong hospital utilisation in relation to life expectancy. DESIGN: Cohort study using a 10% sample (stratified by age and sex) of persons insured by one sickness fund. SETTING: Germany, 1989-1995. SUBJECTS: 69,847 survivors (with a minimum of three more years to live), 1385 persons in last, 1368 in second last, and 1333 in third last year of life. RESULTS: The number of days spent in hospital in the last year of life was lowest for the young (24.2 days under age 25) and the old (23.2 days at age 85+) and was greatest at ages 55-64 (40.6 days). The ratio of days to survivors was highest at age 35-44 (31.0) and fell continuously thereafter to 4.3 at age 85+. Similar patterns were seen for hospital days in the second and third year before death, except that peaks were at 35-44 years (22.5 and 13.7 days respectively). Calculated lifelong number of hospital days increased with age from 54.8 (death at age 20) to 201.0 (age 90). Numbers of hospital days per year of life, averaged over the entire lifespan, were stable at 2.0-2.2 for deaths between age 50 and 90 (and up to 2.7 at age 20). CONCLUSIONS: Lifelong hospital utilisation for persons who die at 50 or later is directly proportional to the number of years lived. These data contradict results from cross sectional studies that suggest an exponential rise in health care costs as longevity increases. They have important implications for projections of future health care expenditure. PMID- 11896138 TI - Psychosocial work environment and myocardial infarction: improving risk estimation by combining two complementary job stress models in the SHEEP Study. AB - OBJECTIVES: Associations between two alternative formulations of job stress derived from the effort-reward imbalance and the job strain model and first non fatal acute myocardial infarction were studied. Whereas the job strain model concentrates on situational (extrinsic) characteristics the effort-reward imbalance model analyses distinct person (intrinsic) characteristics in addition to situational ones. In view of these conceptual differences the hypothesis was tested that combining information from the two models improves the risk estimation of acute myocardial infarction. METHODS: 951 male and female myocardial infarction cases and 1147 referents aged 45-64 years of The Stockholm Heart Epidemiology (SHEEP) case-control study underwent a clinical examination. Information on job stress and health adverse behaviours was derived from standardised questionnaires. RESULTS: Multivariate analysis showed moderately increased odds ratios for either model. Yet, with respect to the effort-reward imbalance model gender specific effects were found: in men the extrinsic component contributed to risk estimation, whereas this was the case with the intrinsic component in women. Controlling each job stress model for the other in order to test the independent effect of either approach did not show systematically increased odds ratios. An improved estimation of acute myocardial infarction risk resulted from combining information from the two models by defining groups characterised by simultaneous exposure to effort-reward imbalance and job strain (men: odds ratio 2.02 (95% confidence intervals (CI) 1.34 to 3.07); women odds ratio 2.19 (95% CI 1.11 to 4.28)). CONCLUSIONS: Findings show an improved risk estimation of acute myocardial infarction by combining information from the two job stress models under study. Moreover, gender specific effects of the two components of the effort-reward imbalance model were observed. PMID- 11896139 TI - Comparing hospital discharge records with death certificates: can the differences be explained? AB - STUDY OBJECTIVE: The quality of mortality statistics is important for epidemiological research. Considerable discrepancies have been reported between death certificates and corresponding hospital discharge records. This study examines whether differences between the death certificate's underlying cause of death and the main condition from the final hospital discharge record can be explained by differences in ICD selection procedures. The authors also discuss the implications of unexplained differences for mortality data quality. DESIGN: Using ACME, a standard software for the selection of underlying cause of death, the compatibility between the underlying cause of death and the final main condition was examined. The study also investigates whether data available in the hospital discharge record, but not reported on the death certificate, influence the selection of the underlying cause of death. SETTING: Swedish death certificates for 1995 were linked to the national hospital discharge register. The resulting database comprised 69 818 people who had been hospitalised during their final year of life. MAIN RESULTS: The underlying cause of death and the main condition differed at Basic Tabulation List level in 54% of the deaths. One third of the differences could not be explained by ICD selection procedures. Adding hospital discharge data changed the underlying cause in 11% of deaths. For some causes of death, including medical misadventures and accidental falls, the effect was substantial. CONCLUSION: Most differences between underlying cause of death and final main condition can be explained by differences in ICD selection procedures. Further research is needed to investigate whether unexplained differences indicate lower data quality. PMID- 11896140 TI - A cancer survival model that takes sociodemographic variations in "normal" mortality into account: comparison with other models. AB - STUDY OBJECTIVES: Sociodemographic differentials in cancer survival have occasionally been studied by using a relative survival approach, where all cause mortality among persons with a cancer diagnosis is compared with that among similar persons without such a diagnosis ("normal" mortality). One should ideally take into account that this "normal" mortality not only depends on age, sex, and period, but also various other sociodemographic variables. However, this has very rarely been done. A method that permits such variations to be considered is presented here, as an alternative to an existing technique, and is compared with a relative survival model where these variations are disregarded and two other methods that have often been used. DESIGN, SETTING, AND PARTICIPANTS: The focus is on how education and marital status affect the survival from 12 common cancer types among men and women aged 40-80. Four different types of hazard models are estimated, and differences between effects are compared. The data are from registers and censuses and cover the entire Norwegian population for the years 1960-1991. There are more than 100 000 deaths to cancer patients in this material. MAIN RESULTS AND CONCLUSIONS: A model for registered cancer mortality among cancer patients gives results that for most, but not all, sites are very similar to those from a relative survival approach where educational or marital variations in "normal" mortality are taken into account. A relative survival approach without consideration of these sociodemographic variations in "normal" mortality gives more different results, the most extreme example being the doubling of the marital differentials in survival from prostate cancer. When neither sufficient data on cause of death nor on variations in "normal" mortality are available, one may well choose the simplest method, which is to model all cause mortality among cancer patients. There is little reason to bother with the estimation of a relative-survival model that does not allow sociodemographic variations in "normal" mortality beyond those related to age, sex, and period. Fortunately, both these less data demanding models perform well for the most aggressive cancers. PMID- 11896141 TI - Milk and coronary heart disease mortality. PMID- 11896142 TI - Kurtoxin, a gating modifier of neuronal high- and low-threshold ca channels. AB - Studies of Ca channels expressed in oocytes have identified kurtoxin as a promising tool for functional and structural studies of low-threshold T-type Ca channels. This peptide, isolated from the venomous scorpion Parabuthus transvaalicus, inhibits low-threshold alpha1G and alpha1H Ca channels expressed in oocytes with relatively high potency and high selectivity. Here we report its effects on Ca channel currents, carried by 5 mm Ba(2+) ions, in rat central and peripheral neurons. In thalamic neurons 500 nm kurtoxin inhibited T-type Ca channel currents almost completely (90.2 +/- 2.5% at -85 mV; n = 6). Its selectivity, however, was less than expected because it also reduced the composite high-threshold Ca channel current recorded in these cells (46.1 +/- 6.9% at -30 mV; n = 6). In sympathetic and thalamic neurons, 250-500 nm kurtoxin partially inhibited N-type and L-type Ca channel currents, respectively. It similarly reduced the high-threshold Ca channel current that remains after a blockade of P-type, N-type, and L-type Ca channels in thalamic neurons. In contrast, kurtoxin facilitated steady-state P-type Ba currents in Purkinje neurons (by 34.9 +/- 3.7%; n = 10). In all cases the kurtoxin effect was voltage dependent and entailed a modification of channel gating. Exposure to kurtoxin slowed current activation kinetics, although its effects on deactivation varied with the channel types. Kurtoxin thus appears as a unique gating-modifier that interacts with different Ca channel types with high affinity. This unusual property and the complex gating modifications it induces may facilitate future studies of gating in voltage-dependent ion channels. PMID- 11896143 TI - The neuronal apoptosis inhibitory protein is a direct inhibitor of caspases 3 and 7. AB - The neuronal apoptosis inhibitory protein (NAIP) was identified as a candidate gene for the inherited neurodegenerative disorder spinal muscular atrophy. NAIP is the founding member of a human protein family that is characterized by highly conserved N-terminal motifs called baculovirus inhibitor of apoptosis repeats (BIR). Five members of the human family of inhibitor of apoptosis proteins including NAIP have been shown to be antiapoptotic in various systems. To date, a mechanism for the antiapoptotic effect of NAIP has not been elucidated. To investigate NAIP function, we found cytoprotection of NAIP-expressing primary cortical neurons treated to undergo caspase-3-dependent apoptosis. The additional treatment of these neurons with the pancaspase inhibitor boc-aspartyl(OMe) fluoromethylketone did not result in increased survival. Similar cytoprotective effects were obtained using HeLa cells transiently transfected with a NAIP N terminal construct and treated to undergo a caspase-3-dependent cell death. To examine whether NAIP inhibits caspases directly, recombinant N-terminal NAIP protein containing BIR domains was overexpressed, purified, and tested for caspase inhibition potential. Our results demonstrate that inhibition of caspases is selective and restricted to the effector group of caspases, with K(i) values as low as approximately 14 nm for caspase-3 and approximately 45 nm for caspase 7. Additional investigations with NAIP fragments containing either one or two NAIP BIRs revealed that the second BIR and to a lesser extent the third BIR alone are sufficient to mediate full caspase inhibition. PMID- 11896144 TI - The NMDA receptor M3 segment is a conserved transduction element coupling ligand binding to channel opening. AB - Ion channels alternate stochastically between two functional states, open and closed. This gating behavior is controlled by membrane potential or by the binding of neurotransmitters in voltage- and ligand-gated channels, respectively. Although much progress has been made in defining the structure and function of the ligand-binding cores and the voltage sensors, how these domains couple to channel opening remains poorly understood. Here we show that the M3 transmembrane segments of the NMDA receptor allosterically interact with both the ligand binding cores and the channel gate. It is proposed that M3 functions as a transduction element whose conformational change couples ligand binding with channel opening. Furthermore, amino acid homology between glutamate receptor M3 segments and the equivalent S6 or TM2 segments in K(+) channels suggests that ion channel activation and gating are both structurally and functionally conserved. PMID- 11896145 TI - Long-term depression in the adult hippocampus in vivo involves activation of extracellular signal-regulated kinase and phosphorylation of Elk-1. AB - Protein kinase cascades likely play a critical role in the signaling events that underlie synaptic plasticity and memory. The extracellular signal-regulated kinase (ERK) cascade is suited well for such a role because its targets include regulators of gene expression. Here we report that the ERK cascade is recruited during long-term depression (LTD) of synaptic strength in area CA1 of the adult hippocampus in vivo and selectively impacts on phosphorylation of the nuclear transcription factor Elk-1. Using a combination of in vivo electrophysiology, biochemistry, pharmacology, and immunohistochemistry, we found the following: (1) ERK phosphorylation, including phosphorylation of nuclear ERK, and ERK phosphotransferase activity are increased markedly, albeit transiently, after the induction of NMDA receptor-dependent LTD at the commissural input to area CA1 pyramidal cells in the hippocampus of anesthetized adult rats; (2) LTD-inducing paired-pulse stimulation fails to produce lasting LTD in the presence of the ERK kinase inhibitor SL327, which suggests that ERK activation is necessary for the persistence of LTD; and (3) ERK activation during LTD results in increased phosphorylation of Elk-1 but not of the transcription factor cAMP response element-binding protein. Our findings indicate that the ERK cascade transduces signals from the synapse to the nucleus during LTD in hippocampal area CA1 in vivo, as it does during long-term potentiation in area CA1, but that the pattern of coupling of the ERK cascade to transcriptional regulators differs between the two forms of synaptic plasticity. PMID- 11896146 TI - Dysfunctional light-evoked regulation of cAMP in photoreceptors and abnormal retinal adaptation in mice lacking dopamine D4 receptors. AB - Dopamine is a retinal neuromodulator that has been implicated in many aspects of retinal physiology. Photoreceptor cells express dopamine D4 receptors that regulate cAMP metabolism. To assess the effects of dopamine on photoreceptor physiology, we examined the morphology, electrophysiology, and regulation of cAMP metabolism in mice with targeted disruption of the dopamine D4 receptor gene. Photoreceptor morphology and outer segment disc shedding after light onset were normal in D4 knock-out (D4KO) mice. Quinpirole, a dopamine D2/D3/D4 receptor agonist, decreased cAMP synthesis in retinas of wild-type (WT) mice but not in retinas of D4KO mice. In WT retinas, the photoreceptors of which were functionally isolated by incubation in the presence of exogenous glutamate, light also suppressed cAMP synthesis. Despite the similar inhibition of cAMP synthesis, the effect of light is directly on the photoreceptors and independent of dopamine modulation, because it was unaffected by application of the D4 receptor antagonist l-745,870. Nevertheless, compared with WT retinas, basal cAMP formation was reduced in the photoreceptors of D4KO retinas, and light had no additional inhibitory effect. The results suggest that dopamine, via D4 receptors, normally modulates the cascade that couples light responses to adenylyl cyclase activity in photoreceptor cells, and the absence of this modulation results in dysfunction of the cascade. Dark-adapted electroretinogram (ERG) responses were normal in D4KO mice. However, ERG b-wave responses were greatly suppressed during both light adaptation and early stages of dark adaptation. Thus, the absence of D4 receptors affects adaptation, altering transmission of light responses from photoreceptors to inner retinal neurons. These findings indicate that dopamine D4 receptors normally play a major role in regulating photoreceptor cAMP metabolism and adaptive retinal responses to changing environmental illumination. PMID- 11896147 TI - Long-lasting potentiation of GABAergic synapses in dopamine neurons after a single in vivo ethanol exposure. AB - The mesolimbic dopamine (DA) system originating in the ventral tegmental area (VTA) is involved in many drug-related behaviors, including ethanol self administration. In particular, VTA activity regulating ethanol consummatory behavior appears to be modulated through GABA(A) receptors. Previous exposure to ethanol enhances ethanol self-administration, but the mechanisms underlying this phenomenon are not well understood. In this study, we examined changes occurring at GABA synapses onto VTA DA neurons after a single in vivo exposure to ethanol. We observed that evoked GABA(A) IPSCs in DA neurons of ethanol-treated animals exhibited paired-pulse depression (PPD) compared with saline-treated animals, which exhibited paired-pulse facilitation (PPF). Furthermore, PPD was still present 1 week after the single exposure to ethanol. An increase in frequency of spontaneous miniature GABA(A) IPSCs (mIPSCs) was also observed in the ethanol treated animals. Additionally, the GABA(B) receptor antagonist (3 aminopropyl)(diethoxymethyl) phosphinic acid shifted PPD to PPF, indicating that presynaptic GABA(B) receptor activation, likely attributable to GABA spillover, might play a role in mediating PPD in the ethanol-treated mice. The activation of adenylyl cyclase by forskolin increased the amplitude of GABA(A) IPSCs and the frequency of mIPSCs in the saline- but not in the ethanol-treated animals. Conversely, the protein kinase A (PKA) inhibitor N-[z-(p bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide significantly decreased both the frequency of spontaneous mIPSCs and the amplitude of GABA(A) IPSCs in the ethanol-treated mice but not in the saline controls. The present results indicate that potentiation of GABAergic synapses, via a PKA-dependent mechanism, occurs in the VTA after a single in vivo exposure to ethanol, and such potentiation might be a key synaptic modification underlying increased ethanol intake. PMID- 11896148 TI - Intrinsic firing dynamics of vestibular nucleus neurons. AB - Individual brainstem neurons involved in vestibular reflexes respond to identical head movements with a wide range of firing responses. This diversity of firing dynamics has been commonly assumed to arise from differences in the types of vestibular nerve inputs to vestibular nucleus neurons. In this study we show that, independent of the nature of inputs, the intrinsic membrane properties of neurons in the medial vestibular nucleus substantially influence firing response dynamics. Hyperpolarizing and depolarizing inputs evoked a markedly heterogenous range of firing responses. Strong postinhibitory rebound firing (PRF) was associated with strong firing rate adaptation (FRA) and occurred preferentially in large multipolar neurons. In response to sinusoidally modulated input current, these neurons showed a pronounced phase lead with respect to neurons lacking strong PRF and FRA. A combination of the hyperpolarization-activated H current and slow potassium currents contributed to PRF, whereas FRA was predominantly mediated by slow potassium currents. An integrate-and-fire-type model, which simulated FRA and PRF, reproduced the phase lead observed in large neurons and showed that adaptation currents were primarily responsible for variations in response phase. We conclude that the heterogeneity of firing dynamics observed in response to head movements in intact animals reflects intrinsic as well as circuit properties. PMID- 11896149 TI - Impaired spatial cognition and synaptic potentiation in a murine model of human immunodeficiency virus type 1 encephalitis. AB - Injection of human immunodeficiency virus type 1 (HIV-1)-infected human monocyte derived macrophages (MDMs) into the basal ganglia of severe combined immunodeficient mice recapitulates histopathologic features of HIV-1 encephalitis (HIVE). Here, we show that the neural damage in HIVE mice extends beyond the basal ganglia and is associated with cognitive impairment. Morris water maze tests showed impaired spatial learning 8 d after MDM injection. Moreover, impaired synaptic potentiation in the hippocampal CA1 subregion was demonstrated at 8 and 15 d. By day 15, post-tetanic, short-term, and long-term potentiation were reduced by 14.1, 29.5, and 45.3% in HIVE mice compared with sham-injected or control animals. Neurofilament (NF) and synaptophysin (SP) antigens were decreased significantly in the CA2 hippocampal subregion of HIVE mice with limited neuronal apoptosis. By day 15, the CA2 region of HIVE mice expressed 3.8- and 2.6-fold less NF and SP than shams. These findings support the notion that HIV-1-infected and immune-competent brain macrophages can cause neuronal damage at distant anatomic sites. Importantly, the findings demonstrate the value of the model in exploring the physiological basis and therapeutic potential for HIV-1 associated dementia. PMID- 11896150 TI - Microtubule-associated protein 1A (MAP1A) and MAP1B: light chains determine distinct functional properties. AB - The microtubule-associated proteins 1A (MAP1A) and 1B (MAP1B) are distantly related protein complexes consisting of heavy and light chains and are thought to play a role in regulating the neuronal cytoskeleton, MAP1B during neuritogenesis and MAP1A in mature neurons. To elucidate functional differences between MAP1B and MAP1A and to determine the role of the light chain in the MAP1A protein complex, we chose to investigate the functional properties of the light chain of MAP1A (LC2) and compare them with the light chain of MAP1B (LC1). We found that LC2 binds to microtubules in vivo and in vitro and induces rapid polymerization of tubulin. A microtubule-binding domain in its NH(2) terminus was found to be necessary and sufficient for these activities. The analysis of LC1 revealed that it too bound to microtubules and induced tubulin polymerization via a crucial but structurally unrelated NH(2)-terminal domain. The two light chains differed, however, in their effects on microtubule bundling and stability in vivo. Furthermore, we identified actin filament binding domains located at the COOH terminus of LC2 and LC1 and obtained evidence that binding to actin filaments is attributable to direct interaction with actin. Our findings establish LC2 as a crucial determinant of MAP1A function, reveal LC2 as a potential linker of neuronal microtubules and microfilaments, and suggest that the postnatal substitution of MAP1B by MAP1A leads to expression of a protein with an overlapping but distinct set of functions. PMID- 11896152 TI - Localization and regulation of the tissue plasminogen activator-plasmin system in the hippocampus. AB - The extracellular protease cascade of tissue plasminogen activator (tPA) and plasminogen has been implicated in neuronal plasticity and degeneration. We show here that unstimulated expression of tPA in the mouse hippocampus is concentrated in the mossy fiber pathway, with little or no expression within the perforant path, the Schaffer collaterals, or neuronal cell bodies. tPA protein is also expressed in vascular endothelial cells throughout the brain parenchyma. Four hours after excitotoxic injury, tPA protein is transiently induced within CA1 pyramidal neurons. The induced CA1 tPA is localized to neurons that survive the injury and is enzymatically active. Within the mossy fiber pathway, injury resulted in decreased tPA protein. In contrast, mossy fiber tPA activity displayed a biphasic character: transient increase at 8 hr, then a decrease by 24 hr after injury. Analysis of plasminogen activator inhibitor-1 (PAI-1) expression showed that PAI-1 antigen is upregulated by 24 hr and could account for the tPA activity downregulation seen at this time point. Plasminogen immunohistochemistry suggested an increase within the mossy fiber pathway after injury. Finally, hippocampal tPA expression among various mammalian species was strikingly different. These results indicate a complex control of tPA protein and enzymatic activity in the hippocampus that may help regulate neuronal plasticity. PMID- 11896151 TI - Estrogen protects against global ischemia-induced neuronal death and prevents activation of apoptotic signaling cascades in the hippocampal CA1. AB - The importance of postmenopausal estrogen replacement therapy in affording protection against the selective and delayed neuronal death associated with cardiac arrest or cardiac surgery in women remains controversial. Here we report that exogenous estrogen at levels that are physiological for hormone replacement in postmenopausal women affords protection against global ischemia-induced neuronal death and prevents activation of apoptotic signaling cascades in the hippocampal CA1 of male gerbils. Global ischemia induced a marked increase in activated caspase-3 in CA1, evident at 6 hr after ischemia. Global ischemia induced a marked upregulation of the proapoptotic neurotrophin receptor p75(NTR) in CA1, evident at 48 hr. p75(NTR) expression was induced primarily in terminal deoxynucleotidyl transferase-mediated UTP nick-end labeling-positive cells, indicating expression in neurons undergoing apoptosis. Global ischemia also induced a marked downregulation of mRNA encoding the AMPA receptor GluR2 subunit in CA1. Caspase-3, p75(NTR), and GluR2 were not significantly changed in CA3 and dentate gyrus, indicating that the ischemia-induced changes in gene expression were cell specific. Exogenous estrogen attenuated the ischemia-induced increases in activated caspase-3 and blocked the increase in p75(NTR) in post-ischemic CA1 neurons but did not prevent ischemia-induced downregulation of GluR2. These findings demonstrate that long-term estrogen at physiological levels ameliorates ischemia-induced hippocampal injury and indicate that estrogen intervenes at the level of apoptotic signaling cascades to prevent onset of death in neurons otherwise "destined to die." PMID- 11896153 TI - Selective blockade of mGlu5 metabotropic glutamate receptors is protective against methamphetamine neurotoxicity. AB - Methamphetamine (MA), a widely used drug of abuse, produces oxidative damage of nigrostriatal dopaminergic terminals. We examined the effect of subtype-selective ligands of metabotropic glutamate (mGlu) receptors on MA neurotoxicity in mice. MA (5 mg/kg, i.p.; injected three times, every 2 hr) induced, 5 d later, a substantial degeneration of striatal dopaminergic terminals associated with reactive gliosis. MA toxicity was primarily attenuated by the coinjection of the noncompetitive mGlu5 receptor antagonists 2-methyl-6-(phenylethynyl)pyridine and (E)-2-methyl-6-styrylpyridine both at 10 mg/kg, i.p.). In contrast, the mGlu1 receptor antagonist 7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (10 mg/kg, i.p.), and the mGlu2/3 receptor agonist (-)-2-oxa-4 aminocyclo[3.1.0]hexane-4,6-dicarboxylic acid (1 mg/kg, i.p.), failed to affect MA toxicity. mGlu5 receptor antagonists reduced the production of reactive oxygen species but did not reduce the acute stimulation of dopamine release induced by MA both in striatal synaptosomes and in the striatum of freely moving mice. We conclude that endogenous activation of mGlu5 receptors enables the development of MA neurotoxicity and that mGlu5 receptor antagonists are neuroprotective without interfering with the primary mechanism of action of MA. PMID- 11896156 TI - Synaptically released glutamate activates extrasynaptic NMDA receptors on cells in the ganglion cell layer of rat retina. AB - NMDA and AMPA receptors (NMDARs and AMPARs) are colocalized at most excitatory synapses in the CNS. Consequently, both receptor types are activated by a single quantum of transmitter and contribute to miniature and evoked EPSCs. However, in amphibian retina, miniature EPSCs in ganglion cell layer neurons are mediated solely by AMPARs, although both NMDARs and AMPARs are activated during evoked EPSCs. One explanation for this discrepancy is that NMDARs are located outside of the synaptic cleft and are activated only when extrasynaptic glutamate levels increase during coincident release from multiple synapses. Alternatively, NMDARs may be segregated at synapses that either are not spontaneously active or yield miniature EPSCs that are too small to detect. In this study, we examined excitatory, glutamatergic synaptic inputs to neurons in the ganglion cell layer of acute slices of rat retina. EPSCs, elicited by electrically stimulating presynaptic bipolar cells, exhibited both NMDAR- and AMPAR-mediated components. However, spontaneous EPSCs exhibited only an AMPAR-mediated component. The effects of low-affinity, competitive receptor antagonists indicated that NMDARs encounter less glutamate than AMPARs during an evoked synaptic response. Reducing glutamate uptake or changing the probability of release preferentially affected the NMDAR component in evoked EPSCs; reducing uptake revealed an NMDAR component in spontaneous EPSCs. These results indicate that NMDARs are located extrasynaptically and that glutamate transporters prevent NMDAR activation by a transmitter released from a single vesicle and limit their activation during evoked responses. PMID- 11896155 TI - Rapid synaptic remodeling by protein kinase C: reciprocal translocation of NMDA receptors and calcium/calmodulin-dependent kinase II. AB - In contrast to the rapid regulation of AMPA receptors, previous evidence has supported the idea that the synaptic density of NMDA-type glutamate receptors is fairly static, modulated only over a long time scale in a homeostatic manner. We report here that selective activation of protein kinase C (PKC) with phorbol esters induces a rapid dispersal of NMDA receptors from synaptic to extrasynaptic plasma membrane in cultured rat hippocampal neurons. PKC activation induced a simultaneous translocation of calcium/calmodulin-dependent kinase II (CaMKII) to synapses but no change in spine number, presynaptic terminal number, or the distribution of AMPA receptors or the synaptic scaffolding protein PSD-95. PKC induced accumulation of CaMKII was dependent on filamentous actin, whereas dispersal of NMDA receptors occurred by a different mechanism independent of actin or CaMKII. Consistent with the decrease in synaptic density of NMDA receptors, phorbol ester pretreatment reduced excitotoxicity. These results reveal a surprisingly dynamic nature to the molecular composition and functional properties of glutamatergic postsynaptic specializations. PMID- 11896154 TI - Expression of a variant form of the glutamate transporter GLT1 in neuronal cultures and in neurons and astrocytes in the rat brain. AB - To identify glutamate transporters expressed in forebrain neurons, we prepared a cDNA library from rat forebrain neuronal cultures, previously shown to transport glutamate with high affinity and capacity. Using this library, we cloned two forms, varying in the C terminus, of the glutamate transporter GLT1. This transporter was previously found to be localized exclusively in astrocytes in the normal mature brain. Specific antibodies against the C-terminal peptides were used to show that forebrain neurons in culture express both GLT1a and GLT1b proteins. The pharmacological properties of glutamate transport mediated by GLT1a and GLT1b expressed in COS-7 cells and in neuronal cultures were indistinguishable. Both GLT1a and GLT1b were upregulated in astrocyte cultures by exposure to dibutyryl cAMP. We next investigated the expression of GLT1b in vivo. Northern blot analysis of forebrain RNA revealed two transcripts of approximately 3 and 11 kb that became more plentiful with developmental age. Immunoblot analysis showed high levels of expression in the cortex, hippocampus, striatum, thalamus, and midbrain. Pre-embedding electron microscopic immunocytochemistry with silver-enhanced immunogold detection was used to localize GLT1b in vivo. In the rat somatosensory cortex, GLT1b was clearly expressed in neurons in presynaptic terminals and dendritic shafts, as well as in astrocytes. The presence of GLT1b in neurons may offer a partial explanation for the observed uptake of glutamate by presynaptic terminals, for the preservation of input specificity at excitatory synapses, and may play a role in the pathophysiology of excitotoxicity. PMID- 11896157 TI - The mitochondrial toxin 3-nitropropionic acid induces striatal neurodegeneration via a c-Jun N-terminal kinase/c-Jun module. AB - Impairments in mitochondrial energy metabolism are thought to be involved in most neurodegenerative diseases, including Huntington's disease (HD). Chronic administration of 3-nitropropionic acid (3-NP), a suicide inhibitor of succinate dehydrogenase, causes prolonged energy impairments and replicates most of the pathophysiological features of HD, including preferential striatal degeneration. In this study, we analyzed one of the mechanisms that could account for this selective 3-NP-induced striatal degeneration. In chronically 3-NP-infused rats, the time course of motor behavioral impairments and histological abnormalities was determined. Progressive alterations of motor performance occurred after 3 d. By histological analysis and terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end-labeling staining, we found a selective neurodegenerescence in the striatum, occurring first in its dorsolateral (DL) part. Activation of c-Jun N-terminal kinase (JNK) was analyzed from brain sections of these rats, using immunocytochemical detection of its phosphorylated form. Activation of JNK occurred progressively and selectively in the DL of the striatum and was followed by c-Jun activation and expression in the same striatal region. To elucidate the role of the JNK/c-Jun module in 3-NP-induced striatal degeneration, we then used primary striatal neurons in culture, in which we replicated neuronal death by application of 3-NP. We found strong nuclear translocation of activated JNK that was rapidly followed by phosphorylation of the transcription factor c-Jun. Overexpression of a dominant negative version of c-Jun, lacking its transactivation domain and phosphorylation sites for activated JNK, completely abolished 3-NP-induced striatal neurodegeneration. We thus conclude that a genetic program controlled by the JNK/c-Jun module is an important molecular event in 3-NP-induced striatal degeneration. PMID- 11896159 TI - Alternative splicing unmasks dendritic and axonal targeting signals in metabotropic glutamate receptor 1. AB - Precise targeting of neurotransmitter receptors to different neuronal compartments is a fundamental step for the establishment and function of synaptic circuitry. Group I metabotropic glutamate receptors, mGluR1 and mGluR5, control glutamatergic neurotransmission by acting both postsynaptically and presynaptically. Four alternatively spliced variants of the mGluR1 gene exist, which differ in their signaling properties and subcellular localization. The present study was undertaken to identify the molecular signals responsible for trafficking of these receptors to different neuronal compartments. Here we report that targeting of mGluR1 to dendrites and axons of transfected retina neurons is controlled by alternative splicing. We have identified in the tail of the receptor a tripeptide motif, which is necessary and sufficient to exclude the splice variant mGluR1b from distal dendrites and to drive it to the axon. This motif, which is present in all the mGluR1 receptors, is masked in mGluR1a by a dominant dendritic signal sequence harbored by the extended C-terminal tail of this splice variant. Furthermore, we show that the identified axonal and dendritic targeting signals are also necessary and sufficient to localize mGluR1b and mGluR1a to the apical and basolateral compartment of Madin-Darby canine kidney cells, respectively, consistent with the existence of common trafficking components for polarized targeting in epithelial cells and neurons. PMID- 11896160 TI - Neuregulin expression at neuromuscular synapses is modulated by synaptic activity and neurotrophic factors. AB - The proper formation of neuromuscular synapses requires ongoing synaptic activity that is translated into complex structural changes to produce functional synapses. One mechanism by which activity could be converted into these structural changes is through the regulated expression of specific synaptic regulatory factors. Here we demonstrate that blocking synaptic activity with curare reduces synaptic neuregulin expression in a dose-dependent manner yet has little effect on synaptic agrin or a muscle-derived heparan sulfate proteoglycan. These changes are associated with a fourfold increase in number and a twofold reduction in average size of synaptic acetylcholine receptor clusters that appears to be caused by excessive axonal sprouting with the formation of new, smaller acetylcholine receptor clusters. Activity blockade also leads to threefold reductions in brain-derived neurotrophic factor and neurotrophin 3 expression in muscle without appreciably changing the expression of these same factors in spinal cord. Adding back these or other neurotrophic factors restores synaptic neuregulin expression and maintains normal end plate band architecture in the presence of activity blockade. The expression of neuregulin protein at synapses is independent of spinal cord and muscle neuregulin mRNA levels, suggesting that neuregulin accumulation at synapses is independent of transcription. These findings suggest a local, positive feedback loop between synaptic regulatory factors that translates activity into structural changes at neuromuscular synapses. PMID- 11896158 TI - Cell cycle proteins exhibit altered expression patterns in lentiviral-associated encephalitis. AB - Cell cycle proteins regulate processes as diverse as cell division and cell death. Recently their role in neuronal death has been reported in several models of neurodegeneration. We have reported previously that two key regulators of the cell cycle, the retinoblastoma susceptibility gene product (pRb) and transcription factor E2F1, exhibit altered immunostaining patterns in simian immunodeficiency virus encephalitis (SIVE). Here we show that E2F1 and the inactivated, hyperphosphorylated form of pRb (ppRb) also exhibit altered immunostaining patterns in human immunodeficiency virus encephalitis (HIVE). Quantification of E2F1 and ppRb staining by immunofluorescent confocal microscopy confirms a significant increase in E2F1 and ppRb in both HIVE and the simian model. This increase in E2F1 and ppRb staining correlates with an increase in the presence of activated macrophages, suggesting a link between changes in cell cycle proteins and the presence of activated macrophages. Changes in ppRb and E2F1 staining in SIVE also correlate with alterations in E2F/DNA binding complexes present in the nuclear and cytoplasmic fractions from both midfrontal cortex and basal ganglia. These findings suggest that changes in cell cycle proteins occur in both HIVE and the simian model and that these changes have functional implications for gene expression in neural cells under encephalitic conditions mediated by macrophage activation or infiltration. PMID- 11896161 TI - Endosomal compartments serve multiple hippocampal dendritic spines from a widespread rather than a local store of recycling membrane. AB - Endosomes are essential to dendritic and synaptic function in sorting membrane proteins for degradation or recycling, yet little is known about their locations near synapses. Here, serial electron microscopy was used to ascertain the morphology and distribution of all membranous intracellular compartments in distal dendrites of hippocampal CA1 pyramidal neurons in juvenile and adult rats. First, the continuous network of smooth endoplasmic reticulum (SER) was traced throughout dendritic segments and their spines. SER occupied the cortex of the dendritic shaft and extended into 14% of spines. Several types of non-SER compartments were then identified, including clathrin-coated vesicles and pits, large uncoated vesicles, tubular compartments, multivesicular bodies (MVBs), and MVB-tubule complexes. The uptake of extracellular gold particles indicated that these compartments were endosomal in origin. Small, round vesicles and pits that did not contain gold were also identified. The tubular compartments exhibited clathrin-coated tips consistent with the genesis of these small, presumably exosomal vesicles. Approximately 70% of the non-SER compartments were located within or at the base of dendritic spines. Overall, only 29% of dendritic spines had endosomal compartments, whereas 20% contained small vesicles. Small vesicles did not colocalize in spines with endosomes or SER. Three-dimensional reconstructions revealed that up to 20 spines shared a recycling pool of plasmalemmal proteins rather than maintaining independent stores at each spine. PMID- 11896162 TI - Versican is upregulated in CNS injury and is a product of oligodendrocyte lineage cells. AB - Chondroitin sulfate proteoglycan (CS-PG) expression is increased in response to CNS injury and limits the capacity for axonal regeneration. Previously we have shown that neurocan is one of the CS-PGs that is upregulated (Asher et al., 2000). Here we show that another member of the aggrecan family, versican, is also upregulated in response to CNS injury. Labeling of frozen sections 7 d after a unilateral knife lesion to the cerebral cortex revealed a clear increase in versican immunoreactivity around the lesion. Western blot analysis of extracts prepared from injured and uninjured tissue also revealed considerably more versican in the injured tissue extract. In vitro studies revealed versican to be a product of oligodendrocyte lineage cells (OLCs). Labeling was seen between the late A2B5-positive stage and the O1-positive pre-oligodendrocyte stage. Neither immature, bipolar A2B5-positive cells, nor differentiated, myelin-forming oligodendrocytes were labeled. The amount of versican in conditioned medium increased as these cells differentiated. Versican and tenascin-R colocalized in OLCs, and coimmunoprecipitation indicated that the two exist as a complex in oligodendrocyte-conditioned medium. Treatment of pre-oligodendrocytes with hyaluronidase led to the release of versican, indicating that its retention at the cell surface is dependent on hyaluronate (HA). In rat brain, approximately half of the versican is bound to hyaluronate. We also provide evidence of a role for CS-PGs in the axon growth-inhibitory properties of oligodendrocytes. Because large numbers of OLCs are recruited to CNS lesions, these results suggest that OLC-derived versican contributes to the inhospitable environment of the injured CNS. PMID- 11896163 TI - cAMP/Ca2+ response element-binding protein function is essential for ocular dominance plasticity. AB - The monocular deprivation model of amblyopia is characterized by a reduction in cortical responses to stimulation of the deprived eye. Although the effects of monocular deprivation on the primary visual cortex have been well characterized physiologically and anatomically, the molecular mechanisms underlying ocular dominance plasticity remain unknown. Previous studies have indicated that the transcription factor adenosine cAMP/Ca(2+) response element-binding protein (CREB) is activated during monocular deprivation. However, it remains unknown whether CREB function is required for the loss of cortical responses to the deprived eye. To address this issue, we used the herpes simplex virus (HSV) to express a dominant negative form of CREB (HSV-mCREB) containing a single point mutation that prevents its activation. Quantitative single-unit electrophysiology showed that cortical expression of this mutated form of CREB during monocular deprivation prevented the loss of responses to the deprived eye. This effect was specific and not related to viral toxicity, because overexpression of functional CREB or expression of beta-galactosidase using HSV injections did not prevent the ocular dominance shift during monocular deprivation. Additional evidence for specificity was provided by the finding that blockade of ocular dominance plasticity was reversible; animals treated with HSV-mCREB recovered ocular dominance plasticity when mCREB expression declined. Moreover, this effect did not result from a suppression of sensory responses caused by the viral infection because neurons in infected cortex responded normally to visual stimulation. These findings demonstrate that CREB function is essential for ocular dominance plasticity. PMID- 11896164 TI - Microglial activation and beta -amyloid deposit reduction caused by a nitric oxide-releasing nonsteroidal anti-inflammatory drug in amyloid precursor protein plus presenilin-1 transgenic mice. AB - 3-4-(2-Fluoro-alpha-methyl-[1,1'-biphenyl]-4-acetyloxy)-3-methoxyphenyl]-2 propenoic acid 4-nitrooxy butyl ester (NCX-2216), a nitric oxide (NO)-releasing derivative of the cyclooxygenase-1-preferring nonsteroidal anti-inflammatory drug (NSAID) flurbiprofen, dramatically reduced both beta-amyloid (Abeta) loads and Congo red staining in doubly transgenic (Tg) amyloid precursor protein plus presenilin-1 mice when administered at 375 ppm in diet between 7 and 12 months of age. This reduction was associated with a dramatic increase in the number of microglia expressing major histocompatibility complex-II antigen, a marker for microglial activation. In contrast, ibuprofen at 375 ppm in diet caused modest reductions in Abeta load but not Congo red staining, suggesting that the effects of this nonselective NSAID were restricted primarily to nonfibrillar deposits. We detected no effects of the cyclooxygenase-2-selective NSAID celecoxib at 175 ppm on amyloid deposition. In short-term studies of 12-month-old Tg mice, we found that the microglia-activating properties of NCX-2216 (7.5 mg small middle dot kg( 1) small middle dot d(-1), s.c.) were present after 2 weeks of treatment. Microglia were not activated by NCX-2216 in non-Tg mice lacking Abeta deposits, nor were microglia activated in Tg animals by flurbiprofen (5 mg small middle dot kg(-1) small middle dot d(-1)) alone. These data are consistent with the argument that activated microglia can clear Abeta deposits. We conclude that the NO generating component of NCX-2216 confers biological actions that go beyond those of typical NSAIDs. In conclusion, NCX-2216 is more efficacious than ibuprofen or celecoxib in clearing Abeta deposits from the brains of Tg mice, implying potential benefit in the treatment of Alzheimer's dementia. PMID- 11896165 TI - Lack of the cell-cycle inhibitor p27Kip1 results in selective increase of transit amplifying cells for adult neurogenesis. AB - The subventricular zone (SVZ) is the largest germinal layer in the adult mammalian brain and comprises stem cells, transit-amplifying progenitors, and committed neuroblasts. Although the SVZ contains the highest concentration of dividing cells in the adult brain, the intracellular mechanisms controlling their proliferation have not been elucidated. We show here that loss of the cyclin dependent kinase inhibitor p27Kip1 has very specific effects on a population of CNS progenitors responsible for adult neurogenesis. Using bromodeoxyuridine and [(3)H]thymidine incorporation to label cells in S phase and cell-specific markers and electron microscopy to identify distinct cell types, we compared the SVZ structure and proliferation characteristics of wild-type and p27Kip1-null mice. Loss of p27Kip1 had no effect on the number of stem cells but selectively increased the number of the transit-amplifying progenitors concomitantly with a reduction in the number of neuroblasts. We conclude that cell-cycle regulation of SVZ adult progenitors is remarkably cell-type specific, with p27Kip1 being a key regulator of the cell division of the transit-amplifying progenitors. PMID- 11896166 TI - Depletion of cholinergic amacrine cells by a novel immunotoxin does not perturb the formation of segregated on and off cone bipolar cell projections. AB - Cone bipolar cells are the first retinal neurons that respond in a differential manner to light onset and offset. In the mature retina, the terminal arbors of On and Off cone bipolar cells terminate in different sublaminas of the inner plexiform layer (IPL) where they form synapses with the dendrites of On and Off retinal ganglion cells and with the stratified processes of cholinergic amacrine cells. Here we first show that cholinergic processes within the On and Off sublaminas of the IPL are present early in development, being evident in the rat on the day of birth, approximately 10 d before the formation of segregated cone bipolar cell axons. This temporal sequence, as well as our previous finding that the segregation of On and Off cone bipolar cell inputs occurs in the absence of retinal ganglion cells, suggested that cholinergic amacrine cells could provide a scaffold for the subsequent in-growth of bipolar cell axons. To test this hypothesis directly, a new cholinergic cell immunotoxin was constructed by conjugating saporin, the ribosome-inactivating protein toxin, to an antibody against the vesicular acetylcholine transporter. A single intraocular injection of the immunotoxin caused a rapid, complete, and selective loss of cholinergic amacrine cells from the developing rat retina. On and Off cone bipolar cells were visualized using an antibody against recoverin, the calcium-binding protein that labels the soma and processes of these interneurons. After complete depletion of cholinergic amacrine cells, cone bipolar cell axon terminals still formed their two characteristic strata within the IPL. These findings demonstrate that the presence of cholinergic amacrine cells is not required for the segregation of recoverin-positive On and Off cone bipolar cell projections. PMID- 11896167 TI - The C domain of netrin UNC-6 silences calcium/calmodulin-dependent protein kinase and diacylglycerol-dependent axon branching in Caenorhabditis elegans. AB - Second messenger systems mediate neuronal responses to extracellular factors that elicit axon branching, turning, and guidance. We found that mutations in Caenorhabditis elegans that affect components of second messenger systems, a G protein subunit, phospholipase Cbeta, diacylglycerol (DAG) kinase, and calcium/calmodulin-dependent protein kinase (CaMKII), have no obvious effect on axon responses to UNC-6 except in animals in which the N-terminal fragment, UNC 6DeltaC, is expressed. In these animals, the mutations enhance or suppress ectopic branching of certain axons. Netrin UNC-6 is an extracellular protein that guides circumferential migrations, and UNC-6DeltaC has UNC-6 guidance activity. We propose that the guidance response elicited by the UNC-6 N-terminal domains involves mechanisms that can induce branching that is sensitive to CaMKII- and DAG-dependent signaling, and that the UNC-6 C domain is required in cis to the N terminal domains to silence the branching and to maintain proper axon morphology. PMID- 11896168 TI - Using optical flow to characterize sensory-motor interactions in a segment of the medicinal leech. AB - Activation of motoneurons innervating leech muscles causes the appearance of a two-dimensional vector field of deformations on the skin surface that can be fully characterized using a new technique (Zoccolan et al., 2001) based on the computation of the optical flow, the two-dimensional vector field describing the point displacements on the skin. These vector fields are characterized by their origin (i.e., the singular point) and by four elementary components that combine linearly: expansion (or compression), rotation, longitudinal shear, and oblique shear. All motoneurons can be classified and recognized according to the components of the deformations they elicit: longitudinal motoneurons give rise almost exclusively to longitudinal negative shear, whereas circular motoneurons give rise to both positive longitudinal shear and significant negative expansion. Oblique motoneurons induce strong oblique shear, in addition to longitudinal shear and negative expansion. Vector fields induced by the contraction of longitudinal, circular, and oblique fibers superimpose linearly. Skin deformations can therefore be attributed rather reliably to the contraction of distinct longitudinal, circular, and oblique muscle fibers. We compared the deformation patterns produced by touching the skin with those produced by intracellular stimulation of P, T, and N cells: vector fields resulting from the activation of P cells were almost identical to those produced by mechanical stimulation. Therefore, motor responses triggered by light or moderate touch are almost entirely mediated by excitation of P cells, with minor contributions from T and N cells. PMID- 11896169 TI - Serotonin release evoked by tail nerve stimulation in the CNS of aplysia: characterization and relationship to heterosynaptic plasticity. AB - Considerable experimental evidence suggests that serotonin (5-HT) at sensory neuron-->motor neuron (SN-->MN) synapses, as well as other neuronal sites, contributes importantly to simple forms of learning such as sensitization and classical conditioning in Aplysia. However, the actual release of 5-HT in the CNS induced by sensitizing stimuli such as tail shock has not been directly demonstrated. In this study, we addressed this question by (1) immunohistochemically labeling central 5-HT processes and (2) directly measuring with chronoamperometry the release of 5-HT induced by pedal tail nerve (P9) shock onto tail SNs in the pleural ganglion and their synapses onto tail MNs in the pedal ganglion. We found that numerous 5-HT-immunoreactive fibers surround both the SN cell bodies in the pleural ganglion and SN axons in the pedal ganglion. Chronoamperometric detection of 5-HT performed with carbon fiber electrodes implanted in the vicinity of tail SN somata and synapses revealed an electrochemical 5-HT signal lasting approximately 40 sec after a brief shock of P9. 5-HT release was restricted to discrete subregions (modulatory fields) of the CNS, including the vicinity of tail SN soma and synapses ipsilateral to the stimulation. Increasing P9 shock frequency augmented the amplitude of the 5-HT signal and, in parallel, increased SN excitability and SN synaptic transmission onto tail MNs. However, the relationship between the amount of 5-HT release and the two forms of SN plasticity was not uniform: SN excitability increased in a graded manner with increased 5-HT release, whereas synaptic facilitation exhibited a highly nonlinear relationship. The development of chronoamperometric techniques in Aplysia now paves the way for a more complete understanding of the contribution of the serotonergic modulatory pathway to memory processing in this system. PMID- 11896171 TI - Activity of thalamic reticular neurons during spontaneous genetically determined spike and wave discharges. AB - This study reports the first intracellular recordings obtained during spontaneous, genetically determined spike and wave discharges (SWDs) in nucleus reticularis thalami (NRT) neurons from the genetic absence epilepsy rats from Strasbourg (GAERS), a model that closely reproduces the typical features of childhood absence seizures. A SWD started with a large hyperpolarization, which was independent of the preceding firing, and decreased in amplitude but did not reverse in polarity up to potentials >/= -90 mV. This hyperpolarization and the slowly decaying depolarization that terminated a SWD were unaffected by recording with KCl-filled electrodes. The prolonged (up to 15 action potentials), high frequency bursts present during SWDs were tightly synchronized between adjacent neurons, correlated with the EEG spike component, and generated by a low threshold Ca(2+) potential, which, in turn, was brought about by the summation of high-frequency, small-amplitude depolarizing potentials. Fast hyperpolarizing IPSPs were not detected either during or in the absence of SWDs. Recordings with KCl-filled electrodes, however, showed a more depolarized resting membrane potential and a higher background firing, whereas the SWD-associated bursts had a longer latency to the EEG spike and a lower intraburst frequency. This novel finding demonstrates that spontaneous genetically determined SWDs occur in the presence of intra-NRT lateral inhibition. The unmasking of these properties in the GAERS NRT confirms their unique association with spontaneous genetically determined SWDs and thus their likely involvement in the pathophysiological processes of the human condition. PMID- 11896170 TI - Episodic bursting activity and response to excitatory amino acids in acutely dissociated gonadotropin-releasing hormone neurons genetically targeted with green fluorescent protein. AB - The gonadotropin-releasing hormone (GnRH) system, considered to be the final common pathway for the control of reproduction, has been difficult to study because of a lack of distinguishing characteristics and the scattered distribution of neurons. The development of a transgenic mouse in which the GnRH promoter drives expression of enhanced green fluorescent protein (EGFP) has provided the opportunity to perform electrophysiological studies of GnRH neurons. In this study, neurons were dissociated from brain slices prepared from prepubertal female GnRH-EGFP mice. Both current- and voltage-clamp recordings were obtained from acutely dissociated GnRH neurons identified on the basis of EGFP expression. Most isolated GnRH-EGFP neurons fired spontaneous action potentials (recorded in cell-attached or whole-cell mode) that typically consisted of brief bursts (2-20 Hz) separated by 1-10 sec. At more negative resting potentials, GnRH-EGFP neurons exhibited oscillations in membrane potential, which could lead to bursting episodes lasting from seconds to minutes. These bursting episodes were often separated by minutes of inactivity. Rapid application of glutamate or NMDA increased firing activity in all neurons and usually generated small inward currents (<15 pA), although larger currents were evoked in the remaining neurons. Both AMPA and NMDA receptors mediated the glutamate-evoked inward currents. These results suggest that isolated GnRH-EGFP neurons from juvenile mice can generate episodes of repetitive burst discharges that may underlie the pulsatile secretion of GnRH, and glutamatergic inputs may contribute to the activation of endogenous bursts. PMID- 11896173 TI - Facilitation of conditioned fear extinction by systemic administration or intra amygdala infusions of D-cycloserine as assessed with fear-potentiated startle in rats. AB - NMDA receptor antagonists block conditioned fear extinction when injected systemically and also when infused directly into the amygdala. Here we evaluate the ability of D-cycloserine (DCS), a partial agonist at the strychnine insensitive glycine-recognition site on the NMDA receptor complex, to facilitate conditioned fear extinction after systemic administration or intra-amygdala infusions. Rats received 10 pairings of a 3.7 sec light and a 0.4 mA footshock (fear conditioning). Fear-potentiated startle (increased startle in the presence vs the absence of the light) was subsequently measured before and after 30, 60, or 90 presentations of the light without shock (extinction training). Thirty non reinforced light presentations produced modest extinction, and 60 or 90 presentations produced nearly complete extinction (experiment 1). DCS injections (3.25, 15, or 30 mg/kg) before 30 non-reinforced light exposures dose-dependently enhanced extinction (experiment 2) but did not influence fear-potentiated startle in rats that did not receive extinction training (experiment 3). These effects were blocked by HA-966, an antagonist at the glycine-recognition site (experiment 4). Neither DCS nor HA-966 altered fear-potentiated startle when injected before testing (experiment 5). The effect of systemic administration was mimicked by intra-amygdala DCS (10 microg/side) infusions (experiment 6). These results indicate that treatments that promote NMDA receptor activity after either systemic or intra-amygdala administration promote the extinction of conditioned fear. PMID- 11896172 TI - Lower sensitivity to stress and altered monoaminergic neuronal function in mice lacking the NMDA receptor epsilon 4 subunit. AB - NMDA receptors, an ionotropic subtype of glutamate receptors (GluRs), play an important role in excitatory neurotransmission, synaptic plasticity, and brain development. They are composed of the GluRzeta subunit (NR1) combined with any one of four GluRepsilon subunits (GluRepsilon1-GluRepsilon4; NR2A-NR2D). Although the GluRzeta subunit exists in the majority of the CNS throughout all stages of development, the GluRepsilon subunits are expressed in distinct temporal and spatial patterns. In the present study, we investigated neuronal functions in mice lacking the embryonic GluRepsilon4 subunit. GluRepsilon4 mutant mice exhibited reductions of [(3)H]MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate] binding and (45)Ca(2+) uptake through the NMDA receptors. The expression of GluRzeta subunit protein, but not GluRepsilon1 and GluRepsilon2 subunit proteins, was reduced in the frontal cortex and striatum of the mutant mice. A postmortem examination in GluRepsilon4 mutant mice revealed that tissue contents of norepinephrine, dopamine, serotonin, and their metabolites were reduced in the hippocampus and that dopamine, as well as serotonin, metabolism was upregulated in the frontal cortex, striatum, hippocampus, and thalamus. To clarify the phenotypical influences of the alteration in neuronal functions, performances in various behavioral tests were examined. GluRepsilon4 mutant mice showed reduced spontaneous locomotor activity in a novel environment and less sensitivity to stress induced by the elevated plus-maze, light-dark box, and forced swimming tests. These findings suggest that GluRepsilon4 mutant mice have dysfunctional NMDA receptors and altered emotional behavior probably caused by changes in monoaminergic neuronal activities in adulthood. PMID- 11896174 TI - Odorant-induced olfactory receptor neural oscillations and their modulation of olfactory bulbar responses in the channel catfish. AB - Peripheral waves (PWs) in the channel catfish are odorant-induced neural oscillations of synchronized populations of olfactory receptor neurons (ORNs) that appear after the initial approximately 500 msec of the response. The mean dominant frequency during the initial 2 sec of PW activity is approximately 28 Hz, declining to approximately 20 Hz in the last sec of a 5 sec stimulus. Recordings of PWs from different regions of a single olfactory lamella and simultaneously from widely separated lamellae within the olfactory organ suggest that PWs are initiated in the sensory epithelium within each olfactory lamella. Simultaneous recordings in vivo from the olfactory organ [electro-olfactogram (EOG) or integrated neural activity], local field potentials (LFPs) from the olfactory bulb (OB), and single and few-unit activity from OB neurons were performed. Cross-correlation analysis of simultaneously recorded odor-induced OB LFPs and either EOG or ORN neural activity showed that oscillations occurring within the OB were lower (<20 Hz) than those of PWs; however, during PW activity, OB LFPs increased both their magnitude and dominant frequencies and became correlated with the PWs. Also during odorant-induced PW activity, the responses of different OB neurons with similar odorant specificity became phase locked to each other and to both the PWs and OB LFPs. PWs are hypothesized to function to strengthen the synaptic transfer of olfactory information at specific glomeruli within the OB. PMID- 11896175 TI - Role of primate substantia nigra pars reticulata in reward-oriented saccadic eye movement. AB - To test the hypothesis that the basal ganglia are related to reward-oriented saccades, we examined activity of substantia nigra pars reticulata (SNr) neurons by using a one-direction-rewarded version of the memory-guided saccade task (1DR). Many SNr neurons changed (decreased or increased) their activity after and before a visual cue (post-cue and pre-cue activity). Post-cue decreases or increases tended to be larger to a contralateral cue. They were often modulated prospectively by the presence or absence of reward, either positively (enhanced in the rewarded condition) or negatively (enhanced in the nonrewarded condition). The positive reward modulation was more common among decreasing type neurons, whereas no such preference was observed among increasing type neurons. The reward contingent decrease in SNr neuronal activity would facilitate rewarded saccades by inducing disinhibition in superior colliculus (SC) neurons. In contrast, the increase in SNr activity would suppress a saccade less selectively (rewarded or nonrewarded) by augmenting inhibition of SC neurons. The post-cue activity was often preceded by anticipatory pre-cue activity. Most typically, post-cue decrease was preceded by pre-cue decrease, selectively when the contralateral side was rewarded. This would reinforce the reward-oriented nature of SNr neuronal activity. The decreases and increases in SNr activity may be derived directly and indirectly, respectively, from the caudate (CD), where neurons show reward-contingent pre-cue and post-cue activity. These results suggest that the CD-SNr-SC mechanism would promote saccades oriented to reward. PMID- 11896176 TI - Stimulus encoding and feature extraction by multiple sensory neurons. AB - Neighboring cells in topographical sensory maps may transmit similar information to the next higher level of processing. How information transmission by groups of nearby neurons compares with the performance of single cells is a very important question for understanding the functioning of the nervous system. To tackle this problem, we quantified stimulus-encoding and feature extraction performance by pairs of simultaneously recorded electrosensory pyramidal cells in the hindbrain of weakly electric fish. These cells constitute the output neurons of the first central nervous stage of electrosensory processing. Using random amplitude modulations (RAMs) of a mimic of the fish's own electric field within behaviorally relevant frequency bands, we found that pyramidal cells with overlapping receptive fields exhibit strong stimulus-induced correlations. To quantify the encoding of the RAM time course, we estimated the stimuli from simultaneously recorded spike trains and found significant improvements over single spike trains. The quality of stimulus reconstruction, however, was still inferior to the one measured for single primary sensory afferents. In an analysis of feature extraction, we found that spikes of pyramidal cell pairs coinciding within a time window of a few milliseconds performed significantly better at detecting upstrokes and downstrokes of the stimulus compared with isolated spikes and even spike bursts of single cells. Coincident spikes can thus be considered "distributed bursts." Our results suggest that stimulus encoding by primary sensory afferents is transformed into feature extraction at the next processing stage. There, stimulus-induced coincident activity can improve the extraction of behaviorally relevant features from the stimulus. PMID- 11896177 TI - Elevated fusiform cell activity in the dorsal cochlear nucleus of chinchillas with psychophysical evidence of tinnitus. AB - Chinchillas with psychophysical evidence of chronic tinnitus were shown to have significantly elevated spontaneous activity and stimulus-evoked responses in putative fusiform cells of the dorsal cochlear nuclei (DCN). Chinchillas were psychophysically trained and tested before and after exposure to a traumatic unilateral 80 dB (sound pressure level) 4 kHz tone. Before exposure, two groups were matched in terms of auditory discrimination performance (noise, and 1, 4, 6, and 10 kHz tones). After exposure, a single psychophysical difference emerged between groups. The exposed group displayed enhanced discrimination of 1 kHz tones (p = 0.00027). Postexposure discrimination of other stimuli was unaffected. It was hypothesized that exposed animals experienced a chronic subjective tone (i.e., tinnitus), resulting from their trauma, and that features of this subjective tone were similar enough to 1 kHz to affect discrimination of 1 kHz objective signals. After psychophysical testing, single-unit recordings were obtained from each animal's DCN fusiform cell layer. Putative fusiform cells of exposed animals showed significantly (p = 0.0136) elevated spontaneous activity, compared with cells of unexposed animals. Putative fusiform cells of exposed animals showed a greater stimulus-evoked response to tones at 1 kHz (p = 0.0000006) and at characteristic-frequency (p = 0.0000009). This increased activity was more pronounced on the exposed side. No increase in stimulus-evoked responses was observed to other frequencies or noise. These parallel psychophysical and electrophysiological results are consistent with the hypothesis that chronic tonal tinnitus is associated with, and may result from, trauma-induced elevation of activity of DCN fusiform cells. PMID- 11896178 TI - Coding and monitoring of motivational context in the primate prefrontal cortex. AB - The prefrontal cortex is involved in acquiring and maintaining information about context, including the set of task instructions and/or the outcome of previous stimulus-response sequences. Most studies on context-dependent processing in the prefrontal cortex have been concerned with such executive functions, but the prefrontal cortex is also involved in motivational operations. We thus wished to determine whether primate prefrontal neurons show evidence of representing the motivational context learned by the monkey. We trained monkeys in a delayed reaction task in which an instruction cue indicated the presence or absence of reward. In random alternation with no reward, the same one of several different kinds of food and liquid rewards was delivered repeatedly in a block of approximately 50 trials, so that reward information would define the motivational context. In response to an instruction cue indicating absence of reward, we found that neurons in the lateral prefrontal cortex not only predicted the absence of reward but also represented more specifically which kind of reward would be omitted in a given trial. These neurons seem to code contextual information concerning which kind of reward may be delivered in following trials. We also found prefrontal neurons that showed tonic baseline activity that may be related to monitoring such motivational context. The different types of neurons were distributed differently along the dorsoventral extent of the lateral prefrontal cortex. Such operations in the prefrontal cortex may be important for the monkey to maximize reward or to modify behavioral strategies and thus may contribute to executive control. PMID- 11896182 TI - Microtubule organization in the green kingdom: chaos or self-order? AB - Plant microtubule arrays differ fundamentally from their animal, fungal and protistan counterparts. These differences largely reflect the requirements of plant composite polymer cell walls and probably also relate to the acquisition of chloroplasts. Plant microtubules are usually dispersed and lack conspicuous organizing centres. The key to understanding this dispersed nature is the identification of proteins that interact with and regulate the spatial and dynamic properties of microtubules. Over the past decade, a number of these proteins have been uncovered, including numerous kinesin-related proteins and a 65 kDa class of structural microtubule-associated proteins that appear to be unique to plants. Mutational analysis has identified MOR1, a probable stabilizer of microtubules that is a homologue of the TOGp-XMAP215 class of high-molecular weight microtubule-associated proteins, and a katanin p60 subunit homologue implicated in the severing of microtubules. The identification of these two proteins provides new insights into the mechanisms controlling microtubule assembly and dynamics, particularly in the dispersed cortical array found in highly polarized plant cells. PMID- 11896183 TI - Musashi: a translational regulator of cell fate. AB - Transcription is thought to have a major role in the regulation of cell fate; the importance of translational regulation in this process has been less certain. Recent findings demonstrate that translational regulation contributes to cell fate specification. The evolutionarily conserved, neural RNA-binding protein Musashi, for example, controls neural cell fate. The protein functions in maintenance of the stem-cell state, differentiation, and tumorigenesis by repressing translation of particular mRNAs. In mammals it might play an important role in activating Notch signalling by repressing translation of the Notch inhibitor m-Numb. PMID- 11896179 TI - Functional interactions between estrogen and insulin-like growth factor-I in the regulation of alpha 1B-adrenoceptors and female reproductive function. AB - The ovarian hormone estradiol (E(2)) and insulin-like growth factor-I (IGF-I) interact in the CNS to regulate neuroendocrine function and synaptic remodeling. Previously, our laboratory showed that 2 d E(2) treatment induces alpha(1B) adrenoceptor expression and promotes IGF-I enhancement of alpha(1)-adrenoceptor potentiation of cAMP accumulation in the preoptic area (POA) and hypothalamus (HYP). This study examined the hypothesis that E(2)-dependent aspects of female reproductive function, including alpha(1B)-adrenoceptor expression and function in the POA and HYP, are mediated by brain IGF-I receptors (IGF-IRs) in female rats. Ovariohysterectomized rats were implanted with a guide cannula aimed at the third ventricle and treated in vivo with vehicle or E(2) daily for 2 d before experimentation. Intracerebroventricular infusions of JB-1, a selective IGF-IR antagonist, were administered every 12 hr beginning 1 hr before the first E(2) injection. Administration of JB-1 during E(2) priming completely blocks hormone induced luteinizing hormone release and partially inhibits hormone-dependent reproductive behavior. Reproductive behavior is restored by intracerebroventricular infusion of 8-bromo-cGMP, the second messenger implicated in alpha(1)-adrenergic facilitation of lordosis. In addition, blockade of IGF-IRs during E(2) priming prevents E(2)-induced increases in alpha(1B)-adenoceptor binding density and abolishes acute IGF-I enhancement of NE-stimulated cAMP accumulation in HYP and POA slices. These data document the existence of a novel mechanism by which IGF-I participates in the remodeling of noradrenergic receptor signaling in the HYP and POA after E(2) treatment. These events may help coordinate the timing of ovulation with the expression of sexual receptivity. PMID- 11896184 TI - Intracellular trafficking of MAN1, an integral protein of the nuclear envelope inner membrane. AB - MAN1 is an integral protein of the inner nuclear membrane that shares the LEM domain, a conserved globular domain of approximately 40 amino acids, with lamina associated polypeptide (LAP) 2 and emerin. Confocal immuofluorescence microscopy studies of the intracellular targeting of truncated forms of MAN1 showed that the nucleoplasmic, N-terminal domain is necessary for inner nuclear membrane retention. A protein containing the N-terminal domain with the first transmembrane segment of MAN1 is retained in the inner nuclear membrane, whereas the transmembrane segments with the C-terminal domain of MAN1 is not targeted to the inner nuclear membrane. The N-terminal domain of MAN1 is also sufficient for inner nuclear membrane targeting as it can target a chimeric type II integral protein to this subcellular location. Deletion mutants of the N-terminal of MAN1 are not efficiently retained in the inner nuclear membrane. When the N-terminal domain of MAN1 is increased in size from approximately 50 kDa to approximately 100 kDa, the protein cannot reach the inner nuclear membrane. Fluorescence recovery after photobleaching experiments of MAN1 fused to green fluorescent protein show that the fusion protein is relatively immobile in the nuclear envelope compared with the endoplasmic reticulum of interphase cells, suggesting binding to a nuclear component. These results are in agreement with the 'diffusion-retention' model for targeting integral proteins to the inner nuclear membrane. PMID- 11896185 TI - F-actin serves as a template for cytokeratin organization in cell free extracts. AB - The microtubule, F-actin, and intermediate filament systems are often studied as isolated systems, yet the three display mutual interdependence in living cells. To overcome limitations inherent in analysis of polymer-polymer interactions in intact cells, associations between these systems were assessed in Xenopus egg extracts. In both fixed and unfixed extract preparations, cytokeratin associated with F-actin cables that spontaneously assembled in the extracts. Time-course experiments revealed that at early time points cytokeratin cables were invariably associated with F-actin cables, while at later time points they could be found without associated F-actin. In extract samples where F-actin assembly was prevented, cytokeratin formed unorganized aggregates rather than cables. Dynamic imaging revealed transport of cytokeratin by moving F-actin as well as examples of cytokeratin release from F-actin. Experimental alteration of F-actin network organization by addition of alpha-actinin resulted in a corresponding change in the organization of the cytokeratin network. Finally, pharmacological disruption of the F-actin network in intact, activated eggs disrupted the normal pattern of cytokeratin assembly. These results provide direct evidence for an association between F-actin and cytokeratin in vitro and in vivo, and indicate that this interaction is necessary for proper cytokeratin assembly after transition into the first mitotic interphase of Xenopus. PMID- 11896186 TI - Evidence for incorporation of free-floating mesothelial cells as a mechanism of serosal healing. AB - Regeneration of the mesothelium is unlike that of other epithelial-like surfaces, as healing does not occur solely by centripetal migration of cells from the wound edge. The mechanism of repair of mesothelium is controversial, but it is widely accepted, without compelling evidence, that pluripotent cells beneath the mesothelium migrate to the surface and differentiate into mesothelial cells. In this study we examined an alternative hypothesis, using in vivo cell-tracking studies, that repair involves implantation, proliferation and incorporation of free-floating mesothelial cells into the regenerating mesothelium. Cultured mesothelial cells, fibroblasts and peritoneal lavage cells were DiI- or PKH26-PCL labelled and injected into rats immediately following mesothelial injury. Implantation of labelled cells was assessed on mesothelial imprints using confocal microscopy, and cell proliferation was determined by proliferating cell nuclear antigen immunolabelling. Incorporation of labelled cells, assessed by the formation of apical junctional complexes, was shown by confocal imaging of zonula occludens-1 protein. Labelled cultured mesothelial and peritoneal lavage cells, but not cultured fibroblasts, implanted onto the wound surface 3, 5 and 8 days after injury. These cells proliferated and incorporated into the regenerated mesothelium, as demonstrated by nuclear proliferating cell nuclear antigen staining and membrane-localised zonula occludens-1 expression, respectively. Furthermore, immunolocalisation of the mesothelial cell marker HBME-1 demonstrated that the incorporated, labelled lavage-derived cells were mesothelial cells and not macrophages as it had previously been suggested. This study has clearly shown that serosal healing involves implantation, proliferation and incorporation of free-floating mesothelial cells into the regenerating mesothelium. PMID- 11896188 TI - Protein phosphatase 4 is required for centrosome maturation in mitosis and sperm meiosis in C. elegans. AB - The centrosome consists of two centrioles surrounded by the pericentriolar material (PCM). In late G2 phase, centrosomes enlarge by recruiting extra PCM, and concomitantly its microtubule nucleation activity increases dramatically. The regulatory mechanisms of this dynamic change of centrosomes are not well understood. Protein phosphatase 4 (PP4) is known to localize to mitotic centrosomes in mammals and Drosophila. An involvement of PP4 in the mitotic spindle assembly has been implicated in Drosophila, but in vivo functions of PP4 in other organisms are largely unknown. Here we characterize two Caenorhabditis elegans PP4 genes, named pph-4.1 and pph-4.2. Inhibition of the function of each gene by RNA-mediated interference (RNAi) revealed that PPH-4.1 was essential for embryogenesis but PPH-4.2 was not. More specifically, PPH-4.1 was required for the formation of spindles in mitosis and sperm meiosis. However, this phosphatase was apparently dispensable for female meiotic divisions, which do not depend on centrosomes. In the cell depleted of pph-4.1 activity, localization of gamma tubulin and a Polo-like kinase homologue to the centrosome was severely disturbed. Immunofluorescence staining revealed that PPH-4.1 was present at centrosomes from prophase to telophase, but not during interphase. These results indicate that PPH-4.1 is a centrosomal protein involved in the recruitment of PCM components to the centrosome, and is essential for the activation of microtubule nucleation potential of the centrosome. Furthermore, chiasmata between homologous chromosomes were often absent in oocytes that lacked pph-4.1 activity. Thus, besides promoting spindle formation, PPH-4.1 appears to play a role in either the establishment or the maintenance of chiasmata during meiotic prophase I. PMID- 11896187 TI - Specific sequences in p120ctn determine subcellular distribution of its multiple isoforms involved in cellular adhesion of normal and malignant epithelial cells. AB - P120 catenin (p120ctn) belongs to the Armadillo family of proteins, which is implicated in cell-cell adhesion and signal transduction. Owing to alternative splicing and multiple translation initiation codons, several p120ctn isoforms can be expressed from a single gene. All p120ctn isoforms share the central Armadillo repeat domain but have divergent N- and C-termini. Little is known about the biological functions of the different isoforms. In this study, we examined the distribution of various p120ctn isoforms and the consequences of their expression in cultured cells of epidermal origin. Immunohistochemical analysis and western blotting revealed that melanocytes and melanoma cells primarily express the long isoform 1A, whereas keratinocytes express shorter isoforms, especially 3A, which localize to cell-cell adhesion junctions in a calcium-dependent manner. The shortest isoform 4A, which was detected in normal keratinocytes and melanocytes, was generally lost from cells derived from squamous cell carcinomas or melanomas. The C-terminal alternatively spliced exon B was present in the p120ctn transcripts in the colon, intestine and prostate, but was lost in several tumor tissues derived from these organs. To test whether p120ctn isoforms serve in distinct biological functions, we transiently transfected the expression constructs into melanoma cells (1205-Lu) and immortalized keratinocytes (HaCaT). Indeed, distinct domains of p120ctn are responsible for its different biological functions. The prominent branching phenotype was induced equally by isoforms 1A, 2A and 3A, whereas the shortest isoform 4A, which was devoid of the N-terminal domain, completely lacked this ability. Also, the exon-B-encoded sequences, as in the isoform 1AB, were sufficient to abolish the branching phenotype as induced by the isoform 1A. The induction of the branching phenotype cosegregated with the nuclear localization of the p120ctn isoforms 1A, 2A and 3A, whereas the isoforms 4A and 1AB, which were excluded from the nucleus, did not induce the branching phenotype. The N-terminal sequences that contain seven out of eight tyrosine residues, recently characterized as potential candidates for phosphorylation by Src kinase, are required for the nuclear localization and for the formation of the branching phenotype. Finally, expression of the p120ctn isoforms, which caused the branching phenotype, was associated with cellular relocalization of E cadherin in HaCaT cells. Collectively, we have identified sequences within the p120ctn N-terminus that are prerequisites for both nuclear localization and the p120ctn-induced branching phenotype. Loss of the cytoplasmic pool of p120ctn from tumor cells suggests an important function for such isoforms in normal cells and tissues. PMID- 11896189 TI - feh-1 and apl-1, the Caenorhabditis elegans orthologues of mammalian Fe65 and beta-amyloid precursor protein genes, are involved in the same pathway that controls nematode pharyngeal pumping. AB - The multigenic family of mammalian Fe65s encodes three highly similar proteins with the same modular organisation: a WW domain and two phosphotyrosine-binding domains. The PTB2 domain of these proteins binds to the cytosolic domains of the Alzheimer's beta-amyloid precursor protein APP and related proteins APLP1 and APLP2, generating a highly redundant system that is hard to dissect by reverse genetics. By searching potential Fe65-like genes in the nematode Caenorhabditis elegans, we identified a single gene, feh-1 (Fe65 homolog-1), encoding a protein with a high sequence similarity to mammalian Fe65s. FEH-1 is also functionally related to mammalian orthologues; in fact its PTB2 domain binds to APL-1, the product of the C. elegans orthologue of APP. Staining with specific antibodies show that the neuromuscular structures of the pharynx are the sites in which FEH 1 is present at highest levels. Expression studies with reporters indicate that the feh-1 gene is also expressed by a subset of the worm neurons. We generated and isolated a deletion allele of feh-1, and the corresponding homozygous mutants arrest as late embryos or as L1 larvae, demonstrating for the first time an essential role for a Fe65-like gene in vivo. The pharynx of homozygous larvae does not contract and the worms cannot feed. Analysis of pharyngeal pumping in heterozygous worms and in feh-1 RNA-interfered worms indicates that dosage of feh 1 function affects the rate of pharyngeal contraction in C. elegans. Interference with apl-1 double-stranded RNA showed a similar effect on pharyngeal pumping, suggesting that FEH-1 and APL-1 are involved in the same pathway. The non redundant system of the nematode will prove useful for studying the basic biology of the Fe65-APP interaction and the molecular events regulated by this evolutionarily conserved system of interacting proteins. PMID- 11896190 TI - Co-regulation of cell adhesion by nanoscale RGD organization and mechanical stimulus. AB - Integrin-mediated cell adhesion is central to cell survival, differentiation and motility. Many cell responses induced by integrins require both receptor occupancy and receptor aggregation, and appear to be regulated by both biochemical and biophysical means. Multidomain extracellular matrix molecules may serve to foster integrin aggregation by presenting local clusters of adhesion ligands, a hypothesis supported by studies with synthetic substrates showing that cell adhesion and migration are enhanced when adhesion ligands are presented in nanoscale clusters. Here, we used a novel synthetic polymer system to present the adhesion ligand GRGDSPK in nanoscale clusters with 1.7, 3.6 or 5.4 peptides per cluster against a non-adhesive background, where the peptide is mobile on a 2 nm polyethylene oxide tether. Average ligand density ranged from 190 to 5270 RGD/microm(2). We used these substrates to study the effects of ligand density and clustering on adhesion of wild-type NR6 fibroblasts, which express alphavbeta3 and alpha5beta1, integrins known to bind to linear RGD peptides. The strength of cell-substratum adhesion was quantified using a centrifugal detachment assay to assess the relative number of cells remaining adherent after a 10 minute application of defined distraction force. An unusual relationship between cell detachment and distraction force at relatively low values of applied force was found on substrates presenting the clustered ligand. Although a monotonic decrease in the number of cells remaining attached would be expected with increasing force on all substrates, we instead observed a peak (adhesion reinforcement) in this profile for certain ligand conditions. On substrates presenting clustered ligands, the fraction of cells remaining attached increased as the distraction force was increased to between 70 and 150 pN/cell, then decreased for higher forces. This phenomenon was only observed on substrates presenting higher ligand cluster sizes (n=3.6 or n=5.4) and was more pronounced at higher ligand densities. Adhesion reinforcement was not observed on fibronectin-coated surfaces. These results support previous studies showing that biophysical cues such as ligand spatial arrangement and extracellular matrix rigidity are central to the governance of cell responses to the external environment. PMID- 11896191 TI - Essential role of human CDT1 in DNA replication and chromatin licensing. AB - Formation of pre-replicative complexes at origins is an early cell cycle event essential for DNA duplication. A large body of evidence supports the notion that Cdc6 protein, through its interaction with the origin recognition complex, is required for pre-replicative complex assembly by loading minichromosome maintenance proteins onto DNA. In fission yeast and Xenopus, this reaction known as the licensing of chromatin for DNA replication also requires the newly identified Cdt1 protein. We studied the role of hCdt1 protein in the duplication of the human genome by antibody microinjection experiments and analyzed its expression during the cell cycle in human non-transformed cells. We show that hCdt1 is essential for DNA replication in intact human cells, that it executes its function in a window of the cell cycle overlapping with pre-replicative complex formation and that it is necessary for the loading of minichromosome maintenance proteins onto chromatin. Intriguingly, we observed that hCdt1 protein, in contrast to other licensing factors, is already present in serum deprived G0 arrested cells and its levels increase only marginally upon re-entry in the cell cycle. PMID- 11896192 TI - Filopodia are conduits for melanosome transfer to keratinocytes. AB - Melanosomes are specialized melanin-synthesizing organelles critical for photoprotection in the skin. Melanosome transfer to keratinocytes, which involves whole organelle donation to another cell, is a unique biological process and is poorly understood. Time-lapse digital movies and electron microscopy show that filopodia from melanocyte dendrites serve as conduits for melanosome transfer to keratinocytes. Cdc42, a small GTP-binding protein, is known to mediate filopodia formation. Melanosome-enriched fractions isolated from human melanocytes expressed the Cdc42 effector proteins PAK1 and N-WASP by western blotting. Expression of constitutively active Cdc42 (Cdc42(V12)) in melanocytes co-cultured with keratinocytes induced a highly dendritic phenotype with extensive contacts between melanocytes and keratinocytes through filopodia, many of which contained melanosomes. These results suggest a unique role for filopodia in organelle transport and, in combination with our previous work showing the presence of SNARE proteins and rab3a on melanosomes, suggest a novel model system for melanosome transfer to keratinocytes. PMID- 11896193 TI - Cytoplasmic dynein-associated structures move bidirectionally in vivo. AB - Intracellular organelle transport is driven by motors that act upon microtubules or microfilaments. The microtubulebased motors, cytoplasmic dynein and kinesin, are believed to be responsible for retrograde and anterograde transport of intracellular cargo along microtubules. Many vesicles display bidirectional movement; however, the mechanism regulating directionality is unresolved. Directional movement might be accomplished by alternative binding of different motility factors to the cargo. Alternatively, different motors could associate with the same cargo and have their motor activity regulated. Although several studies have focused on the behavior of specific types of cargoes, little is known about the traffic of the motors themselves and how it correlates with cargo movement. To address this question, we studied cytoplasmic dynein dynamics in living Dictyostelium cells expressing dynein intermediate chain-green fluorescent protein (IC-GFP) fusion in an IC-null background. Dynein-associated structures display fast linear movement along microtubules in both minus-end and plus-end directions, with velocities similar to that of dynein and kinesin-like motors. In addition, dynein puncta often rapidly reverse their direction. Dynein stably associates with cargo moving in both directions as well as with those that rapidly reverse their direction of movement, suggesting that directional movement is not regulated by altering motor-cargo association but rather by switching activity of motors associated with the cargo. These observations suggest that both plus- and minus-end-directed motors associate with a given cargo and that coordinated regulation of motor activities controls vesicle directionality. PMID- 11896194 TI - Low-energy laser irradiation promotes the survival and cell cycle entry of skeletal muscle satellite cells. AB - Low energy laser irradiation (LELI) has been shown to promote skeletal muscle cell activation and proliferation in primary cultures of satellite cells as well as in myogenic cell lines. Here, we have extended these studies to isolated myofibers. These constitute the minimum viable functional unit of the skeletal muscle, thus providing a close model of in vivo regeneration of muscle tissue. We show that LELI stimulates cell cycle entry and the accumulation of satellite cells around isolated single fibers grown under serum-free conditions and that these effects act synergistically with the addition of serum. Moreover, for the first time we show that LELI promotes the survival of fibers and their adjacent cells, as well as cultured myogenic cells, under serum-free conditions that normally lead to apoptosis. In both systems, expression of the anti-apoptotic protein Bcl-2 was markedly increased, whereas expression of the pro-apoptotic protein BAX was reduced. In culture, these changes were accompanied by a reduction in the expression of p53 and the cyclin-dependent kinase inhibitor p21, reflecting the small decrease in viable cells 24 hours after irradiation. These findings implicate regulation of these factors as part of the protective role of LELI against apoptosis. Taken together, our findings are of critical importance in attempts to improve muscle regeneration following injury. PMID- 11896195 TI - Quantitation and functional characterization of neural cells derived from ES cells using nestin enhancer-mediated targeting in vitro. AB - To gain insight into early events of neurogenesis, transgenic embryonic stem (ES) cells were generated using the enhanced green fluorescence protein (EGFP) reporter gene under the regulatory control of the neural stem cell marker, nestin. The expression of EGFP in undifferentiated ES cells suggested that the onset of endogenous nestin occurred before neurulation. Upon differentiation of ES cells, the EGFP expression became confined to the neural lineage and asynchrony in ES-cell-derived neural differentiation was evident. The EGFP intensity was prominent in the proliferative progenitors and unipolar neurons, whereas downregulation occurred in differentiating bi- and multipolar neurons. This was corroborated quantitatively using flow cytometry where maximal generation of neural progenitors was observed 4-12 days post-plating. The proliferative potential of neural progenitors as well as glia, in contrast to post-mitotic neurons, was also evident by time-lapse microscopy. The functional characterization of progenitors revealed an absence of voltage-activated inward currents, whereas the Na+ current (INa) was detected prior to Ca2+ currents (ICa) in differentiating neurons. Additionally, inhibitory receptor-operated channels could be detected at these early stages of development in bi- and multipolar neurons suggesting that the pre-committed progenitors had retained their intrinsic ability to give rise to functional neurons. This study sheds new light on early events of neurogenesis defining the quantitative and qualitative aspects and demarcating the functional neural cell types from ES cells in vitro. PMID- 11896196 TI - Vascular endothelial cells that express dystroglycan are involved in angiogenesis. AB - We have earlier shown that dystroglycan (DG) is a lamininbinding protein and as such is a cell adhesion molecule. DG is a heterodimer of alpha and beta DG subunits. beta-dystroglycan (betaDG) is the membrane spanning subunit, whereas the alpha subunit is bound to the extracellular domain of betaDG. To study physiological function of the protein, we expressed full-length DG (FL-DG) and the cytoplasmic domain of betaDG (deltabetaDG) in bovine aortic endothelial cells (BAE) and examined their effects on cell adhesion, migration, proliferation and tube formation. FL-DG enhanced adhesion of BAE to laminin-1, whereas deltabetaDG inhibited it. Cell migration was inhibited by overexpressing deltabetaDG in these cells, although it was not affected by FL-DG overexpression. In a proliferation assay, FL-DG decreased the growth rate, and it also caused cells to extensively spread. deltabetaDG caused opposite effects; it increased the growth rate and reduced the cell surface area. In a tube formation assay on Matrigel, FL-DG caused an obvious inhibition, whereas deltabetaDG accelerated tube formation. These results suggest a potential role of endothelial dystroglycan in the control of angiogenesis. Anti-betaDG immunohistochemistry indicated increased expression of DG in vascular endothelial cells within malignant tumors. By contrast, the antibody failed to stain endothelial cells in normal tissues. In cultured BAE, the level of betaDG was low in serum-deprived quiescent cells and was upregulated in proliferating cells. Our results suggest that the expression of DG in endothelial cells is under a dynamic regulation and may play a role in angiogenesis. PMID- 11896197 TI - GIT1 functions in a motile, multi-molecular signaling complex that regulates protrusive activity and cell migration. AB - GIT1 is a multidomain protein that is thought to function as an integrator of signaling pathways controlling vesicle trafficking, adhesion and cytoskeletal organization. It regulates ARF GTPases and has binding domains for paxillin and PIX, which is a PAK-binding protein and an exchange factor for Rac. We show that GIT1 cycles between at least three distinct subcellular compartments, including adhesion-like structures, the leading edge and cytoplasmic complexes. The cytoplasmic structures, which also contain paxillin, PAK and PIX, do not detectably co-localize with endosomal Golgi or membrane markers, suggesting that they represent a novel supramolecular complex. The GIT1 cytoplasmic complexes are motile and tended to move toward the cell periphery where they joined existing adhesions. In retracting regions of the cells, the GIT1 complexes moved away from the disassembling adhesions toward the cell body. Using deletion mutants, we have identified domains that target GIT1 to each of the compartments. Localization to adhesions and the leading edge requires the paxillin-binding domain, which comprises the C-terminal 140 residues (cGIT1), whereas targeting to the cytoplasmic complexes requires the central region that contains ankyrin repeats and the PIX-binding domain. Expression of GIT1 or cGIT, but not nGIT1 in which the paxillin-binding domain is deleted, increases the rate of migration and the size and number of protrusions. The latter are inhibited when GIT1 is co expressed with a kinase-dead PAK, suggesting that the GIT1 interaction with PAK is required for enhanced migration and protrusive activity. Furthermore, GIT1 targets constitutively activated PAK to adhesions and the leading edge via its interaction with paxillin. Since expression of cGIT targets endogenous GIT1 to the leading edge, it appears that the leading edge is the location of GIT1 responsible for these activities. Thus, GIT1 is a component of a motile, multimolecular complex that traffics a set of signaling components to specific locations in the cell where they regulate localized activities. PMID- 11896198 TI - Analysis of Chlamydomonas SF-assemblin by GFP tagging and expression of antisense constructs. AB - Striated fiber assemblin (SF-assemblin or SFA) is the major component of the striated microtubule-associated fibers (SMAFs) in the flagellar basal apparatus of green flagellates. We generated nuclear transformants of Chlamydomonas expressing green fluorescent protein (GFP) fused to the C-terminus of SFA. SFA GFP assembled into striated fibers that exceeded those of wild-type cells in size by several fold. At elevated temperatures (>or=32 degrees C) SFA-GFP was mostly soluble and heat shock depolymerized the SMAFs. C-terminal deletions of 18 or only six residues disturbed the ability of SFA-GFP to polymerize, indicating an important role of the C-terminal domain for fiber formation. The exchange of the penultimate Ser275 with alanine made SFA-GFP highly insoluble, causing aberrant fiber formation and conferring heat stability to the fibers. By contrast, a replacement with glutamic acid increased the solubilty of the molecule, indicating that phosphorylation on Ser275 might control solubility of SFA. In vivo observation of GFP fluorescence showed that SFA-GFP fibers were disassembled during mitosis. In cells overexpressing full-length or truncated SFA-GFP, the amount of wild-type protein was reduced. Elevated temperatures dissolved SFA-GFP fibers and induced the synthesis of SFA, suggesting that cells control both the amount of soluble and polymeric SFA. By expressing constructs consisting of cDNA and genomic DNA for parts of SFA in antiparallel configuration, the amount of SFA was severely reduced. In these strains we observed defects in flagellar assembly, indicating an important role for noncontractile striated roots in the flagella apparatus. PMID- 11896199 TI - The projection domain of MAP2b regulates microtubule protrusion and process formation in Sf9 cells. AB - The expression of microtubule-associated protein 2 (MAP2), developmentally regulated by alternative splicing, coincides with neurite outgrowth. MAP2 proteins contain a microtubule-binding domain (C-terminal) that promotes microtubule assembly and a poorly characterized domain, the projection domain (N terminal), extending at the surface of microtubules. MAP2b differs from MAP2c by an additional sequence of 1372 amino acids in the projection domain. In this study, we examined the role of the projection domain in the protrusion of microtubules from the cell surface and the subsequent process formation in Sf9 cells. In this system, MAP2b has a lower capacity to induce process formation than MAP2c. To investigate the role of the projection domain in this event, we expressed truncated forms of MAP2b and MAP2c that have partial or complete deletion of their projection domain in Sf9 cells. Our results indicate that process formation is induced by the microtubule-binding domain of these MAP2 proteins and is regulated by their projection domain. Furthermore, the microtubule-binding activity of MAP2b and MAP2c truncated forms as well as the structural properties of the microtubule bundles induced by them do not seem to be the only determinants that control the protrusion of microtubules from the cell surface in Sf9 cells. Rather, our data suggest that microtubule protrusion and process formation are regulated by intramolecular interactions between the projection domain and its microtubule-binding domain in MAP2b. PMID- 11896200 TI - Partner telomeres during anaphase in crane-fly spermatocytes are connected by an elastic tether that exerts a backward force and resists poleward motion. AB - As chromosomes move polewards during anaphase in crane-fly spermatocytes, trailing arms commonly stretch backwards for a brief time, as if tethered to their partners. To test that notion, a laser microbeam was used to sever trailing arms and thereby release telomere-containing arm segments (called acentric fragments because they lack kinetochores) from segregating chromosomes. Analysis of the movement of acentric fragments after their release provided clear evidence that previously conjoined partners were indeed tethered at their telomeres and that tethers exerted backward forces that were sufficient to move the fragment across the equator and into the opposite half-spindle. To address concerns that tethers might be artifacts of in vitro cell culture, spermatocytes were fixed in situ, and stretched arms within fixed cells provided strong evidence for tethers in vivo. The substantial resistance that tethers impose on the poleward movement of chromosomes must normally be over-ridden by the poleward 'pulling' forces exerted at kinetochores. In spermatocytes, poleward forces are supplied primarily by the 'traction fibers' that are firmly attached to kinetochores through end-on attachments to the plus ends of kinetochore microtubules. PMID- 11896201 TI - DNA ligase I null mouse cells show normal DNA repair activity but altered DNA replication and reduced genome stability. AB - DNA ligase I is the key ligase for DNA replication in mammalian cells and has also been reported to be involved in a number of recombination and repair processes. Our previous finding that Lig1 knockout mouse embryos developed normally to mid-term before succumbing to a specific haematopoietic defect was difficult to reconcile with a report that DNA ligase I is essential for the viability of cultured mammalian cells. To address this issue, we generated a second Lig1 targeted allele and found that the phenotypes of our two Lig1 mutant mouse lines are identical. Widely different levels of Lig1 fusion transcripts were detected from the two targeted alleles, but we could not detect any DNA ligase I protein, and we believe both are effective Lig1 null alleles. Using foetal liver cells to repopulate the haematopoietic system of lethally irradiated adult mice, we demonstrate that the haematopoietic defect in DNA-ligase-I deficient embryos is a quantitative deficiency relating to reduced proliferation rather than a qualitative block in any haematopoietic lineage. DNA ligase I null fibroblasts from Lig1 mutant embryos showed an accumulation of DNA replication intermediates and increased genome instability. In the absence of a demonstrable deficiency in DNA repair we postulate that, unusually, genome instability may result directly from the DNA replication defect. Lig1 null mouse cells performed better in the survival and replication assays than a human LIG1 point mutant, and we suggest that the complete absence of DNA ligase I may make it easier for another ligase to compensate for DNA ligase I deficiency. PMID- 11896203 TI - HIV-1 envelope glycoprotein 120 increases intercellular adhesion molecule-1 expression by human endothelial cells. AB - Human immunodeficiency virus type 1 (HIV-1) infection is often associated with central nervous system damage and vascular complications. However, the mechanisms of this association are largely unknown. We examined the effect of HIV-1 envelope glycoprotein 120 (gp120) on cell adhesion molecule expression by endothelial cells. We found, for the first time, that both soluble and membrane-bound gp120 could significantly increase the expression of human endothelial intercellular adhesion molecule-1 (ICAM-1) at both mRNA and protein levels, but not vascular cell adhesion molecule-1 and E-selectin. The specificity of gp120-mediated response was demonstrated by blocking experiments using a specific monoclonal antibody against gp120, which successfully abolished the gp120-mediated increase of ICAM-1 expression. Furthermore, there was a significant increase of human monocytic cell line THP-1 adherence onto the gp120-treated endothelial monolayers. This increased cell adhesion was effectively blocked by either anti gp120 or anti-ICAM antibodies. These findings suggest that HIV-1 gp120-mediated endothelial ICAM-1 expression could be one of the important mechanisms of HIV-1 pathogenesis. PMID- 11896205 TI - Frameshift mutations of human gastrin receptor gene (hGARE) in gastrointestinal cancers with microsatellite instability. AB - Gastrointestinal tumors with DNA mismatch repair (MMR) defects show microsatellite instability (MSI) and harbor frameshift mutations in coding mononucleotide repeats of cancer-related genes (targets). We assessed MSI status in 233 sporadic gastrointestinal tumors. We classified as MSI-H (high-frequency microsatellite instability) 15 (10%) of 150 colorectal cancers and 13 (16%) of 83 gastric cancers. We searched for frameshift mutations in a coding poly(T)(8) tract within the gastrin receptor gene (hGARE), which has a potential role in gastrointestinal carcinogenesis. To this purpose, we screened 43 unstable tumors (including 15 hereditary nonpolyposis colorectal cancer cases previously classified as MSI-H), 98 stable tumors, as well as 3 MMR-deficient and 4 MMR proficient gastrointestinal cancer cell lines. We found mutations in 8 (19%) of the 43 MSI-H tumors but in none of the 98 stable cancers. hGARE mutation frequency was similar in gastric (23%) and colorectal cancers, including sporadic (13%) and hereditary (20%) cases. All mutated tumors proved to harbor frameshift mutations in other cancer-related genes that are considered as targets in MSI tumorigenesis. The MMR-deficient and gastrin-sensitive LoVo colorectal cancer cells also showed a hGARE heterozygous frameshift mutation, but expressed only the mutated allele. All detected mutations can be predicted to generate a truncated protein carrying amino acid changes. On the basis of genetic findings, we propose hGARE as a new candidate target gene in MSI tumorigenesis. Functional studies are warranted to elucidate the mechanism by which the hGARE mutation might contribute to gastrointestinal carcinogenesis. PMID- 11896204 TI - Gene mutations in lymphoproliferative disorders of T and NK/T cell phenotypes developing in renal transplant patients. AB - Post-transplantation lymphoproliferative disorder (PT-LPD) is characterized by lymphoid proliferation after organ or bone marrow transplantation. In Western countries, most cases of PT-LPD are B-cell-derived and Epstein-Barr virus associated, in which alterations of c-myc, p53, and N-ras genes might play a role in disease progression. In Japan, PT-LPD of T- and NK/T-cell types are not uncommon in renal transplant patients. Mutations of p53 (exons 4 through 8), K ras (exons 1 and 2), c-kit (exons 11 and 17), and beta-catenin genes (exon 3) in 12 cases of these diseases were analyzed by PCR single strand conformation polymorphism and then by direct sequencing. p53 gene mutations were detected in 5 of 5 cases of peripheral T-cell lymphoma, 3 (60%) of 5 cases of adult T-cell leukemia/lymphoma, and 1 of 2 cases of NK/T cell lymphoma. Twenty-five percent of T and NK/T cell lymphomas showed K-ras mutations. Mutations of c-kit and beta catenin genes were found in 33% of cases. Among a total of 42 substitution mutations, 40 were transitions with involvement of CpG sites in 20 to 30% of cases. Most cases had at least one mutation that changed an amino acid, which might have provided the selection pressure for expansion. These findings suggested that p53 gene mutations might play a central role in development of peripheral T-cell lymphoma including adult T-cell leukemia/lymphoma in renal transplant patients. PMID- 11896206 TI - ABCA1 mRNA and protein distribution patterns predict multiple different roles and levels of regulation. AB - Mutations in ABCA1 cause the allelic disorders familial hypolipoproteinemia and Tangier Disease. To identify where ABCA1 was likely to have a functional role, we determined the cellular and tissue-specific patterns of murine ABCA1 expression. RT-PCR and Western blot analysis on dissected murine tissues demonstrated broad expression of ABCA1 mRNA and protein in many tissues with prominent protein expression in liver, testis, and adrenal tissue. In situ hybridization and immunohistochemistry experiments demonstrated specific patterns of ABCA1 expression at the cellular level, with hepatocytes, the epithelial lining of the digestive system and bladder, the proximal convoluted tubule of the kidney, and Purkinje and cortical pyramidal neurons containing abundant ABCA1 protein. Significant discordance between relative mRNA and protein expression patterns suggests the possibility of post-transcriptional regulation of ABCA1 expression in selected cells or tissues. We also show that ABCA1 protein levels are up regulated specifically in the liver after exposure to an atherogenic diet for 7 days, supporting a major role for the liver in dietary modulation of HDL-C levels. Our observations show that ABCA1 is expressed in a pattern consistent with its role in HDL-C metabolism. Additionally, ABCA1 may have important functional roles in other cell types independent of HDL-C regulation. PMID- 11896207 TI - Promoter methylation and silencing of PTEN in gastric carcinoma. AB - The PTEN/MMAC1/TEP1 gene (phosphatase and tensin homolog deleted on chromosome 10/mutated in multiple advanced cancers/TGF-beta regulated and epithelial cell enriched phosphatase 1), which regulates the signaling pathways of Akt, is a novel tumor suppressor gene implicated in multiple cancers. Because a number of tumor suppressor genes are known to be silenced by aberrant promoter methylation, we examined the methylation status of the 5' CpG islands of PTEN using methylation-specific PCR. The altered expression of PTEN in 310 gastric carcinomas was analyzed by immunohistochemical staining using tissue-array and clinicopathologic profiles related to PTEN expression were characterized. Of 310 consecutive gastric carcinomas, 62 cases (20%) showed expression loss of PTEN. Altered PTEN expression was significantly associated with tumor depth and size, lymphatic invasion, advanced stage, pTNM stage, and patient survival (p < 0.001). The promoter methylation frequency of PTEN was found to be present in 26 (39%) of 66 cases examined, and 19 (73%) of 26 gastric cancer tissues showing promoter methylation exhibited the loss of PTEN expression. Abnormalities in the expression of PTEN significantly correlated with promoter methylation (p < 0.001). In conclusion, silencing of the PTEN gene occurs frequently in gastric carcinoma and aberrant promoter methylation is a major mechanism of silencing of the PTEN gene. The abnormalities of the PTEN gene are associated with tumor progression, metastasis, and survival. PMID- 11896208 TI - Analysis of intrapulmonary vessels and epithelial-endothelial interactions in the human developing lung. AB - The establishment of a sufficiently wide and functional blood-gas interface is of critical importance in lung development, but development of the intrapulmonary vascular system including alveolar capillary vessels still remains unclear. In this study, we first characterized the structural development of the vascular system in accordance with that of airways in human fetal lungs at the pseudoglandular phase (8, 13, and 16 weeks gestation) by examining the immunohistochemical distribution of CD34 and alpha-smooth muscle actin (SMA). Using double immunohistochemistry and 3-dimensional reconstruction techniques, endothelial cells in the developing lung could be classified into two different types according to the characteristics of their adjacent cells (presence or absence of SMA-positive cells) and their distribution (proximal or distal lung parenchyme). Endothelial cells without SMA-positive cells developed into a capillary network surrounding the budding components of distal airways during the mid-pseudoglandular phase before communicating with proximal vessels. We then examined the immunoreactivity of thrombomodulin and von Willebrand factor (vWF) in endothelial cells. Endothelial cells of the capillary network were mainly positive for vWF during the early gestational stages, but altered their phenotypes to those of mature lungs (vWF negative and thrombomodulin positive) during the terminal sac phase. We subsequently determined the immunohistochemical distribution of vascular endothelial growth factor (VEGF). Epithelial cells of the most distal airways were intensely positive for VEGF. These results suggest that VEGF present in airway epithelial cells is involved in the maturation as well as proliferation of capillary endothelial cells. Epithelial-endothelial interactions during lung development are considered very important in the establishment of the functional blood-gas interface. PMID- 11896209 TI - Kinetics of CD11b/CD18 up-regulation during infection with the agent of human granulocytic ehrlichiosis in mice. AB - The agent of human granulocytic ehrlichiosis (aoHGE) is a tick-borne, obligate intracellular, granulocytotropic bacterium able to infect numerous host species. Given its unique niche and the leukopenia often noted with infection, we investigated the effect of acute aoHGE infection on neutrophil activation by evaluating surface expression of the beta 2 integrin CD11b/CD18 in a mouse model using FACS analysis. Infection resulted in neutrophil activation with up regulation of CD11b/CD18 in multiple strains of mice, however, hematologic analysis showed no apparent role for CD11b/CD18 in mediating peripheral leukopenia. Because IFN-gamma is an important cytokine during granulocytic ehrlichiosis and is known to activate leukocytes, we investigated the potential role of IFN-gamma in CD11b/CD18 up-regulation. Neutrophils from IFN-gamma knock out mice became activated during aoHGE infection, however, the kinetics of activation differed from wild-type mice. In addition, activation correlated directly with the presence of bacteria because neutrophils with large intracytoplasmic morula also expressed higher levels of CD11b/CD18. CD11b/CD18 seemed to be critical to early bacterial clearance and killing in vivo because infection of mice with targeted genetic disruption of CD11b/CD18 resulted in an initial increase in bacterial burden compared with wild-type mice. Similarly, in vitro culture of neutrophils from infected CD11b/CD18 knock-out mice resulted in a marked increase in bacterial proliferation compared with congenic controls. The data support crucial roles of CD11b/CD18 and IFN-gamma-mediated cell activation as mechanisms that limit bacterial replication. PMID- 11896210 TI - CD44 is exposed to the extracellular matrix at invasive sites in basal cell carcinomas. AB - We have previously shown, by light microscopy, that the level of expression of CD44 (pan-CD44, CD44v3, CD44v5, and CD44v6) in human basal cell carcinomas is related to growth pattern and invasiveness (Br J Dermatol 1099;140:17-25). We have now studied the fine distribution of these CD44 isoforms in the same tumors using immunoelectron microscopy. Despite the strong differences in the level of expression in tumor areas with different growth patterns, CD44 was consistently found almost exclusively at intercellular surfaces, with a very strong predilection for widened intercellular pouches, ie, identical to the distribution in the normal epidermis. This prevalent distribution corroborates a role for CD44 in maintaining hyaluronan-filled spaces (J Histochem Cytochem 1998;46:241-248). However, the correlation between the presence of CD44 and the presence of such pouches was not absolute, indicating that other factors are involved as well. In contrast to the prevailing literature, we also found a weak but distinct labeling of cell surfaces facing the extracellular matrix. Interestingly, this appeared significantly elevated in the thinnest, most irregular, and usually most peripheral tumor cell strands, where it was associated with tumor cell protrusions and absence of a basal lamina. Thus, the CD44(+) protrusions were in direct contact with the extracellular matrix and apparently represented sites of invasion. The mechanisms that may contribute to a role of CD44 at these sites include binding of extracellular matrix components (notably hyaluronan) and several biologically active factors such as hepatocyte growth factor/scatter factor and matrix metalloproteinase 9. PMID- 11896211 TI - Immortal activated human hepatic stellate cells generated by ectopic telomerase expression. AB - Telomere shortening controls the entry of cells into senescence. Functional expression of the telomerase catalytic subunit (human telomerase reverse transcriptase or hTERT) stabilizes telomere length and extends the life span of various normal human cells. Our aim was to assess the role of telomerase activity and telomere maintenance in regulating the proliferation of activated human hepatic stellate cells (HSCs), to establish an immortal human HSC cell line. Human HSCs were isolated from surgical specimens of normal liver and infected with a retrovirus expressing hTERT. Ectopic expression of hTERT reconstituted telomerase activity and maintained telomere length in human HSCs. Control human HSCs, which were either not infected or infected with a retroviral vector containing only the neomycin resistance gene, showed no detectable telomerase activity and had slightly shortened telomeres. These telomerase-negative HSCs entered a nondividing state after about 9 to 15 passages and senesced. In contrast, telomerase-positive HSCs to date have undergone 69 passages. Telomerase positive HSCs did not undergo oncogenic transformation and exhibit morphologic and functional characteristics of activated HSCs. Microarray and RT-PCR analysis showed that mRNA expression patterns in telomerase-positive HSCs are very similar to those in activated human HSCs. Plating telomerase-positive HSCs on a basement membrane-like matrix reverts them toward a more quiescent phenotype. In conclusion, introduction of hTERT into activated human HSCs immortalizes them and maintains their activated phenotype. This newly developed cell line will be a useful tool to study the cell biology of human HSCs in culture. PMID- 11896212 TI - Infantile dilated X-linked cardiomyopathy, G4.5 mutations, altered lipids, and ultrastructural malformations of mitochondria in heart, liver, and skeletal muscle. AB - Mutations in the Xq28 gene G4.5 lead to dilated cardiomyopathy (DCM). Differential splicing of G4.5 results in a family of proteins called "tafazzins" with homology to acyltransferases. These enzymes assemble fatty acids into membrane lipids. We sequenced G4.5 in two kindreds with X-linked DCM and in two unrelated men, one with idiopathic DCM and the other with DCM of arrhythmogenic right ventricular dysplasia. We examined the ultrastructure of heart, liver, and muscle biopsy specimens in these three DCM types; we used gas chromatography to compare fatty acid composition in heart, liver, and muscle autopsy specimens of two patients of kindred 1 with that of controls. In X-linked DCM, G4.5 had a stop codon (E188X), a nonsense mutation, in kindred 1 and an amino acid substitution (G240R), a missense mutation, in kindred 2. In the two men with isolated DCM, G4.5 was not mutated. Ultrastructural mitochondrial malformations were present in the biopsy tissues of the patients with DCM. Cardiac biopsy specimens of both kindreds with X-linked DCM exhibited greatly enlarged mitochondria with large bundles of stacked, compacted, disarrayed cristae that differed from those of the two types of isolated DCM. Autopsy tissue of patients with X-linked DCM had decreased unsaturated and increased saturated fatty acid concentrations. Seven of 13 published G4.5 missense mutations, including the one presented here, occur in acyltransferase motifs. Impaired acyltransferase function could result in increased fatty acid saturation that would decrease membrane fluidity. Mitochondrial membrane proliferation may be an attempt to compensate for impaired function of acyltransferase. Cardiac ultrastructure separates X-linked DCM with G4.5 mutations from the two types of isolated DCM without G4.5 mutations. Electron microscopy of promptly fixed myocardial biopsy specimens has a role in defining the differential diagnosis of DCM. Mutational analysis of the G4.5 gene also serves this purpose. PMID- 11896213 TI - Nestin as a marker for proliferative endothelium in gliomas. AB - Nestin is one of the intermediate filaments abundantly produced in the developing central nervous system and somites in the embryonic stage. Nestin is also reportedly detected in gliomas/glioblastomas. We retested nestin expression in brain tumors having a range of malignancy grades using immunostaining. The intensity of nestin immunostaining roughly paralleled the malignancy grade of the gliomas. However, many tumors were negative for nestin immunostaining, while nestin immunostaining was invariably detected in tumor endothelium regardless of glioma malignancy grades or brain tumor types. We suspected that angiogenic epithelial cells may express nestin, and we found that nestin was highly positive in bovine aortic endothelial cells in static culture. However, nestin expression decreased when the endothelial cells underwent laminar shear stress flow, under which endothelial cells exhibit differentiated features and a decreased rate of growth. Because nestin is highly expressed in growing endothelial cells, we examined its expression in hemangioblastomas because hemangioblasts are thought to be a precursor for angiogenic epithelial cells. As expected, nestin immunostained strongly in all four samples of hemangioblastomas. We suggest that nestin is not only a marker for neuroepithelial stem cells and glioma cells but also for tumor endothelial cells during rapid growth. PMID- 11896214 TI - NF1 tumor suppressor mRNA is targeted to the cell-cell contact zone in Ca(2+) induced keratinocyte differentiation. AB - SUMMARY: We have previously shown that NF1 (type 1 neurofibromatosis) p21ras GTPase-activating tumor suppressor protein undergoes major relocalization during the formation of cell-cell junctions in differentiating keratinocytes in vitro. This prompted us to study the distribution of NF1 mRNA under the same conditions by in situ hybridization. In differentiating keratinocytes, the NF1 mRNA signal intensified within the cell cytoplasm within the first 0.5 to 2 hours after induction of cellular differentiation. First, the hybridization signal was evenly distributed throughout the cytoplasm. Subsequently, NF1 mRNA was gradually polarized to the cellular periphery at the side of cell-cell junctions and finally disappeared. Reappearance of NF1 mRNA was found in migrating keratinocytes forming a bilayered culture. Disruption of microfibrillar cytoskeleton, but not microtubules, caused a marked change in the subcellular distribution of NF1 mRNA. This data may suggest that intact actin microfilaments are essential for transport of NF1 mRNA to the cell periphery. This is the first study demonstrating that NF1, or any tumor suppressor mRNA, belongs to a rare group of mRNAs not targeted to free polysomes or ribosomes of the rough endoplasmic reticulum. This finding recognizes a potential way for post transcriptional modification of NF1 expression. PMID- 11896215 TI - MR imaging of ligament and tendon injuries of the fingers. AB - Magnetic resonance (MR) imaging can provide important information for diagnosis and evaluation of soft-tissue trauma in the fingers. An optimal imaging technique should include proper positioning, dedicated surface coils, and specific protocols for the suspected abnormalities. Familiarity with the fine anatomy of the normal finger is crucial for identifying pathologic entities. MR imaging is a powerful method for evaluating acute and chronic lesions of the stabilizing articular elements (volar plate and collateral ligaments) of the fingers and thumbs, particularly in the frequently affected proximal interphalangeal and metacarpophalangeal joints. As in other body regions, MR imaging is also useful for depicting traumatic conditions of the extensor and flexor tendons, including injuries to the pulley system. In general, normal ligaments and tendons have low signal intensity on MR images, whereas disruption manifests as increased signal intensity. Radiologists need to understand the full spectrum of finger abnormalities and associated MR imaging findings. PMID- 11896216 TI - Progressive bone and joint abnormalities of the spine and lower extremities in cerebral palsy. AB - Bone and joint changes in cerebral palsy result from muscle spasticity and contracture. The spine and the joints of the lower extremity are most commonly affected. Scoliosis may progress rapidly and may continue after skeletal maturity. Increased thoracic kyphosis and lumbar lordosis, spondylolisthesis, spondylolysis, and pelvic obliquity may accompany the scoliosis. Progressive hip flexion and adduction lead to windswept deformity, increased femoral anteversion, apparent coxa valga, subluxation, deformity of the femoral head, hip dislocation, and formation of a pseudoacetabulum. In the knee, flexion contracture, patella alta, and patellar fragmentation are the most commonly seen abnormalities. Recurvatum deformity can also develop in the knee secondary to contracture of the rectus femoris muscle. Progressive equinovalgus and equinovarus of the foot and ankle are associated with rocker-bottom deformity and subluxation of the talonavicular joint. Early recognition of progressive deformity in patients with cerebral palsy allows timely treatment and prevention of irreversible change. PMID- 11896217 TI - US of ductal carcinoma in situ. AB - Little is known about the ultrasonographic (US) features of ductal carcinoma in situ (DCIS) of the breast because this entity usually manifests as pure mammographic calcifications and is rarely evaluated with US. US findings were recorded in 70 patients with DCIS and then analyzed and correlated with mammographic and histologic findings. A microlobulated mass with mild hypoechogenicity, ductal extension, and normal acoustic transmission was the most common US finding in DCIS. Spiculated margins, marked hypoechogenicity, a thick echogenic rim, and posterior acoustic shadowing at US often suggested the presence of invasion. US performed with a 10-13-MHz transducer and optimal technique can be used as a complement to mammography in detecting and evaluating DCIS of the breast, as it demonstrates breast lesions associated with malignant microcalcifications in most cases. The main benefit of identifying a US abnormality in women with mammographically detected DCIS is to allow the use of US to guide interventional procedures (eg, needle biopsy, needle localization). US may also be helpful in detecting DCIS without calcifications and in evaluating disease extent in women with dense breasts. Nevertheless, further research is needed to delineate the role of US in the evaluation of patients with DCIS. PMID- 11896219 TI - CT and MR imaging features of adnexal torsion. AB - In adnexal torsion, the ovary, ipsilateral fallopian tube, or both twist with the vascular pedicle, resulting in vascular compromise. Unrelieved torsion is likely to cause hemorrhagic infarction as the degree of arterial occlusion increases. Therefore, early diagnosis is important to preserve the affected ovary. Adnexal torsion commonly accompanies an ipsilateral ovarian neoplasm or cyst but can also occur in normal ovaries, usually in children. Although ultrasonography is typically the initial emergent examination, computed tomography (CT) and magnetic resonance (MR) imaging may also be useful diagnostic tools. Common CT and MR imaging features of adnexal torsion include fallopian tube thickening, smooth wall thickening of the twisted adnexal cystic mass, ascites, and uterine deviation to the twisted side. Uncommon imaging findings in adnexal torsion that are specific to hemorrhagic infarction include hemorrhage in the thickened fallopian tube, hemorrhage within the twisted ovarian mass, and hemoperitoneum. Additional imaging findings that can suggest hemorrhagic infarction include eccentric smooth wall thickening exceeding 10 mm in a cystic ovarian mass converging on the thickened fallopian tube and lack of contrast enhancement of the internal solid component or thickened wall of the twisted ovarian mass. Early diagnosis can help prevent irreversible structural damage and may allow conservative, ovary-sparing treatment. PMID- 11896220 TI - Practical MR imaging of female pelvic floor weakness. AB - Pelvic floor weakness is common in middle-aged and elderly parous women and is often associated with stress incontinence, uterine prolapse, constipation, and incomplete defecation. Most patients with incontinence and minimal pelvic floor weakness can be treated based on physical examination and basic urodynamic findings. However, in women with symptoms of multicompartment involvement for whom a complex repair is planned or who have undergone previous repairs, magnetic resonance (MR) imaging can be a useful preoperative planning tool. The MR imaging evaluation is performed with the patient in the supine position, without contrast agents, and within 15 minutes. A multicoil array and a rapid half-Fourier T2 weighted imaging sequence are used to obtain sagittal images while the patient is at rest and during pelvic strain, followed by axial images. On these images, the radiologist identifies the pubococcygeal line (which represents the level of the pelvic floor), the H and M lines (which are helpful for confirming pelvic floor laxity), and the angle of the levator plate with the pubococcygeal line (which is helpful for identifying small bowel prolapse). In the appropriate patient, MR images provide relatively easy three-dimensional conceptualization of the pelvic floor and can significantly influence treatment planning. PMID- 11896221 TI - Management of visceral interventional radiology catheters: a troubleshooting guide for interventional radiologists. AB - Visceral interventional radiology catheters can be difficult to exchange or remove for a variety of reasons. These reasons include exit of the guide wire through the side holes of the catheter, blockage of the catheter, difficulty unlocking the pigtail, retention of the string after catheter removal, migration of the string ahead of the guide wire, catheter fracture, and snaring of an adjacent stent by the pigtail. Secure fixation of the catheter to the skin is important. A technique that allows secure fixation without direct puncture and suturing of the catheter to the skin is recommended. If a catheter falls out or is inadvertently removed, access can occasionally be regained and the catheter can be replaced without repuncture. The timing of catheter removal is based on the clinical condition of the patient and the daily output from the catheter. "Tractography" is a useful study before removal of any catheter that requires a mature tract for removal, particularly cholecystostomy catheters and transpleural catheters. In biliary catheter exchange, the most vital issue is the position of the side holes of the catheter. If an abscess cavity remains large after catheter drainage, the catheter can be repositioned or a second catheter can be placed. PMID- 11896222 TI - Postoperative anatomic and pathologic findings at CT following gastrectomy. AB - Helical computed tomography (CT) is useful in identifying postoperative anatomic changes, complications, and tumor recurrence in gastric cancer patients who have undergone gastrectomy. Postoperative anatomic changes can usually be identified on consecutive CT scans. Complications include anastomotic leakage, duodenal stump leakage, intraabdominal bleeding, wound complications, and other less common complications (postoperative pancreatitis, retention of surgical foreign bodies, diffuse peritonitis). The degree and extent of bowel wall thickening is important in diagnosing tumor recurrence; however, CT lacks specificity. Large or conglomerated lymph node metastases can be easily diagnosed at CT; however, small solitary or focal metastases may not be detected or differentiated from nonmetastatic nodes. Ascites, a common finding with peritoneal seeding in gastrointestinal tumors, is well depicted at CT. Hematogenous metastases from gastric carcinoma are most frequently seen in the liver and are best demonstrated with helical CT performed during the portal venous phase of enhancement (sensitivity >90% for the detection of lesions >1 cm). The sophisticated surgical procedures used in gastrectomy can alter normal anatomy and make image interpretation difficult; thus, familiarity with the appearance of postoperative anatomic changes, complications, and tumor recurrence is essential for accurate CT evaluation of affected patients. PMID- 11896223 TI - MR imaging and CT of vascular anomalies and connections in patients with congenital heart disease: significance in surgical planning. AB - To plan effective management of congenital heart disease, one needs the clearest understanding of the anatomy. Although echocardiography and angiography are the dominant imaging modalities in patients with congenital heart disease, magnetic resonance (MR) imaging and computed tomography (CT) are valuable noninvasive adjuncts. MR imaging and CT are effective in demonstrating the complex cardiovascular morphology present in congenital heart disease, especially the extracardiac morphology. In patients with tetralogy of Fallot with complex pulmonary artery anatomy, MR imaging and CT are useful in demonstrating the pulmonary artery anatomy, along with the significant aortopulmonary collateral vessels. In the heterotaxy syndromes, patients often have unusual atriovenous connections. MR imaging allows accurate identification of the hepatic, systemic, and pulmonary veins and their relationships to both atria. CT and MR are the imaging modalities of choice in a patient who is thought to have a vascular ring. Treatment of aortic coarctation is usually performed on the basis of typical clinical and echocardiographic findings. In patients with atypical clinical or echocardiographic findings, MR imaging and CT yield helpful information that can change the treatment plan. The enhanced preoperative understanding of congenital heart disease provided by MR imaging and CT simplifies surgical decision making and consequently may improve outcome. PMID- 11896226 TI - Imaging of Merkel cell carcinoma. AB - Merkel cell carcinoma (MCC) is a rare, highly malignant cutaneous tumor, primarily of the head and neck, that requires timely diagnosis, adequate staging, and aggressive therapy. MCC tends to be overlooked in the early stage, has a high propensity for invading local and regional nodal basins, and exhibits a high postoperative recurrence rate, with distant thoracic, abdominal, and central nervous system metastases. Conventional radiography and cross-sectional imaging show lesions similar to those originating from other small cell carcinomas. Nuclear medicine procedures such as sentinel node scintigraphy, somatostatin receptor scintigraphy, and positron emission tomography may be used to supplement judicious cross-sectional imaging evaluation, thereby adding diagnostic value in staging and providing therapeutic guidance. Ultimately, however, the diagnosis relies exclusively on pathologic findings at immunohistochemical staining and electron microscopy. The rarity of MCC and the resulting insufficient awareness of this neoplasm often delay correct identification and treatment, which essentially consists of wide-margin surgical excision of the primary tumor and local and regional radiation therapy. To date, clinical information is still insufficient to fully appreciate the role of imaging in MCC management. A better imaging algorithm is expected with increased awareness and improved clinical understanding of this uncommon skin neoplasm. PMID- 11896225 TI - Bronchial carcinoid tumors of the thorax: spectrum of radiologic findings. AB - Bronchial carcinoid tumors are neuroendocrine neoplasms that range from low-grade typical carcinoids to more aggressive atypical carcinoids and therefore demonstrate a wide spectrum of clinical behaviors and histologic features. Typical and atypical bronchial carcinoids have similar imaging features. Because most bronchial carcinoids are located in central airways, radiologic findings are usually related to bronchial obstruction. Central bronchial carcinoids manifest as an endobronchial nodule or hilar or perihilar mass with a close anatomic relationship to the bronchus. The mass is usually a well-defined, round or ovoid lesion and may be slightly lobulated at radiography and computed tomography (CT). Associated atelectasis, air trapping, obstructing pneumonitis, and mucoid impaction may also be seen. Peripheral bronchial carcinoids appear as solitary nodules. Calcification is common and is easily visualized at CT. Bronchial carcinoids demonstrate high signal intensity on T2-weighted and short-inversion time inversion recovery magnetic resonance images. Prognosis of bronchial carcinoids is highly dependent on histologic findings: Atypical carcinoids have certain features that suggest a more aggressive nature. Typical bronchial carcinoids generally have an excellent prognosis, whereas atypical bronchial carcinoids have a worse prognosis. Therefore, understanding the histologic, clinical, and radiologic features of bronchial carcinoids facilitates accurate diagnosis and helps optimize surgical planning. PMID- 11896227 TI - Paleoradiology: advanced CT in the evaluation of nine Egyptian mummies. AB - Axial thin-collimation state-of-the-art spiral computed tomography (CT) was combined with sagittal and coronal reformatting, three-dimensional (3D) reconstruction, and virtual "fly-through" techniques to nondestructively study nine Egyptian mummies. These techniques provided important paleopathologic and historical information about mummification techniques, depicted anatomy in the most informative imaging plane, illustrated the soft-tissue preservation and physical appearance of mummies in superb detail, and generated an intriguing virtual tour through hollow mummified remains without harming the specimens themselves. Images generated with these methods can help archaeologists and Egyptologists understand these fascinating members of mankind and can serve as adjunct visual aids for laypersons who are interested in mummies. CT has emerged as the imaging modality of choice for the examination of Egyptian mummies due to its noninvasive cross-sectional nature and inherently superior contrast and spatial resolution. As multi-detector row CT and postprocessing tools evolve, the capabilities and applications of CT will continue to proliferate, attesting to the expanded versatility and utility of CT as a noninvasive research tool in the multidisciplinary study of Egyptian mummies. PMID- 11896229 TI - From the archives of the AFIP. Benign tumors and tumorlike lesions of the gallbladder and extrahepatic bile ducts: radiologic-pathologic correlation. Armed Forces Institute of Pathology. AB - A diverse spectrum of benign tumors and tumorlike lesions arises from the gallbladder and bile ducts, and despite their diversity, these lesions share common embryologic origins and histologic characteristics. Although these lesions are relatively uncommon, their importance lies in their ability to mimic malignant lesions in these locations. Benign neoplasms are derived from the epithelial and nonepithelial structures that compose the normal gallbladder and bile ducts. The epithelium gives rise to adenomas, cystadenomas, and the unusual condition of biliary papillomatosis. Granular cell tumors, neurofibromas, ganglioneuromas, paragangliomas, and leiomyomas are examples of benign tumors that may originate from nonepithelial structures. Tumorlike lesions are more commonly found in the gallbladder and include xanthogranulomatous cholecystitis, adenomyomatous hyperplasia, cholesterol polyps, and heterotopias. In the clinical setting of a patient with nonspecific abdominal complaints or symptoms of biliary obstruction, the discovery of a gallbladder or bile duct polyp or mass, gallbladder wall thickening, or biliary stricture is most often indicative of malignancy. However, the differential diagnosis should include benign tumors and tumorlike lesions. The preoperative determination of a benign lesion may significantly alter therapy and patient prognosis. PMID- 11896230 TI - Best cases from the AFIP. Juvenile polyposis of the stomach. Armed Forces Institute of Pathology. PMID- 11896232 TI - Medical image segmentation with knowledge-guided robust active contours. AB - Medical image segmentation techniques typically require some form of expert human supervision to provide accurate and consistent identification of anatomic structures of interest. A novel segmentation technique was developed that combines a knowledge-based segmentation system with a sophisticated active contour model. This approach exploits the guidance of a higher-level process to robustly perform the segmentation of various anatomic structures. The user need not provide initial contour placement, and the high-level process carries out the required parameter optimization automatically. Knowledge about the anatomic structures to be segmented is defined statistically in terms of probability density functions of parameters such as location, size, and image intensity (eg, computed tomographic [CT] attenuation value). Preliminary results suggest that the performance of the algorithm at chest and abdominal CT is comparable to that of more traditional segmentation techniques like region growing and morphologic operators. In some cases, the active contour-based technique may outperform standard segmentation methods due to its capacity to fully enforce the available a priori knowledge concerning the anatomic structure of interest. The active contour algorithm is particularly suitable for integration with high-level image understanding frameworks, providing a robust and easily controlled low-level segmentation tool. Further study is required to determine whether the proposed algorithm is indeed capable of providing consistently superior segmentation. PMID- 11896231 TI - Computer-assisted analysis of three-dimensional MR angiograms. AB - The software tools required for postprocessing of magnetic resonance (MR) angiograms include the following functions: data handling, image visualization, and vascular analysis. A custom postprocessing software called Magnetic Resonance Angiography Computer Assisted Analysis (MARACAS) has been developed. This software combines the most commonly used three-dimensional visualization techniques with image processing methods for analysis of vascular morphology on MR angiograms. The main contributions of MARACAS are (a) implementation of a fast method for stenosis quantification on three-dimensional MR angiograms, which is clinically applicable in a personal computer-based system; and (b) portability to the most widespread platforms. The quantification is performed in three steps: extraction of the vessel centerline, detection of vessel boundaries in planes locally orthogonal to the centerline, and calculation of stenosis parameters on the basis of the resulting contours. Qualitative results from application of the method to data from patients showed that the vessel centerline correctly tracked the vessel path and that contours were correctly estimated. Quantitative results obtained from images of phantoms showed that the computation of stenosis severity was accurate. PMID- 11896233 TI - Multiparametric color-encoded brain MR imaging in Talairach space. AB - Clinical magnetic resonance (MR) imaging of the brain is typically performed in the standard three orthogonal planes of the magnet, with little regard to head positioning. Multiple sequences with different imaging parameters are performed, and gray-scale images are obtained and displayed separately. The authors have implemented, and currently advocate, the routine acquisition of coregistered transverse images after roll, yaw, and pitch correction to Talairach space. Talairach, anterior commissure (AC)-posterior commissure (PC) referenced, stereotactic space has been widely embraced by the neuroscience community. This standardization should lead to more reproducible and readily interpretable MR examinations. A method is described to obtain direct AC-PC referenced (Talairach space) MR images. Sample protocols are provided. Coregistered T1-weighted, T1 weighted with contrast material administration, T2-weighted, fluid-attenuated inversion recovery (FLAIR), MR angiography, fractional anisotropy, and functional MR imaging sequences are presented, depicting a wide range of imaging parameters applied to normal brain anatomy in Talairach space. Illustrative examples of pathology are also provided. Color encoding is discussed and exploited to display and integrate multiparameter MR imaging contrast and white-matter-tract direction (anisotropy). The color composites may reduce the number of images needed for review by a factor of three or four and facilitate interpretation. PMID- 11896234 TI - Novel Internet-based tool for correcting apparent sensitivity and specificity of diagnostic tests to adjust for referral (verification) bias. AB - Referral (verification) bias-the selective sampling of a population under evaluation for definitive confirmation of disease status-has been recognized as affecting the measured sensitivity and specificity of diagnostic tests. The authors developed an Internet-based Java applet to correct the apparent (measured) values of sensitivity and specificity of diagnostic tests to adjust for referral (verification) bias. The applet was applied to the diagnosis of coronary artery disease by means of exercise stress testing. Referral rates for coronary arteriography can be adjusted separately for patients with positive and negative test results. The more complicated situation, in which the results are stratified in terms of exercise heart rate, was also investigated. PMID- 11896236 TI - Sea-level fingerprinting as a direct test for the source of global meltwater pulse IA. AB - The ice reservoir that served as the source for the meltwater pulse IA remains enigmatic and controversial. We show that each of the melting scenarios that have been proposed for the event produces a distinct variation, or fingerprint, in the global distribution of meltwater. We compare sea-level fingerprints associated with various melting scenarios to existing sea-level records from Barbados and the Sunda Shelf and conclude that the southern Laurentide Ice Sheet could not have been the sole source of the meltwater pulse, whereas a substantial contribution from the Antarctic Ice Sheet is consistent with these records. PMID- 11896235 TI - Outbreak of poliomyelitis in Hispaniola associated with circulating type 1 vaccine-derived poliovirus. AB - An outbreak of paralytic poliomyelitis occurred in the Dominican Republic (13 confirmed cases) and Haiti (8 confirmed cases, including 2 fatal cases) during 2000-2001. All but one of the patients were either unvaccinated or incompletely vaccinated children, and cases occurred in communities with very low (7 to 40%) rates of coverage with oral poliovirus vaccine (OPV). The outbreak was associated with the circulation of a derivative of the type 1 OPV strain, probably originating from a single OPV dose given in 1998-1999. The vaccine-derived poliovirus associated with the outbreak had biological properties indistinguishable from those of wild poliovirus. PMID- 11896237 TI - Solution of a 20-variable 3-SAT problem on a DNA computer. AB - A 20-variable instance of the NP-complete three-satisfiability (3-SAT) problem was solved on a simple DNA computer. The unique answer was found after an exhaustive search of more than 1 million (2(20)) possibilities. This computational problem may be the largest yet solved by nonelectronic means. Problems of this size appear to be beyond the normal range of unaided human computation. PMID- 11896238 TI - Superconductivity in a spin-ladder cuprate. PMID- 11896239 TI - Chemical ecology: missed opportunities? PMID- 11896240 TI - Appointment pending. Zerhouni seems headed for NIH, and new scrap over stem cells. PMID- 11896241 TI - Appointment pending. A spin-off with vision. PMID- 11896242 TI - Stem cell research. Studies cast doubt on plasticity of adult cells. PMID- 11896243 TI - Graduate training. South Korea scrambles to fill Ph.D. slots. PMID- 11896244 TI - Superconductivity. New observations give stripes theory a lift. PMID- 11896245 TI - India: Academic science gets big boost in budget. PMID- 11896247 TI - Astronomy. Two satellites get new lease on life. PMID- 11896246 TI - Air pollution. Small particles add up to big disease risk. PMID- 11896248 TI - Spain. New cancer center makes a big splash. PMID- 11896250 TI - Astrophysics. Stellar pair whirls in a 5-minute dash. PMID- 11896249 TI - Neuroscience. Neurons weigh options, come to a decision. PMID- 11896251 TI - Public health. U.S. vaccine supply falls seriously short. PMID- 11896252 TI - Public health. Gates Foundation rearranges public health universe. PMID- 11896253 TI - Public health. Dead virus walking. PMID- 11896254 TI - Public health. Is live smallpox lurking in the Arctic? PMID- 11896255 TI - Public health. 'Destructionists' fight to keep a dream alive. PMID- 11896256 TI - Gunter Wachtershauser profile. Between a rock and a hard place. PMID- 11896257 TI - Ecology. Soil fertility and hunger in Africa. PMID- 11896258 TI - Ecology. Seeking new recruits. PMID- 11896260 TI - Planetary disks. A dusty business. PMID- 11896259 TI - Environmental chemistry. Tracking hexavalent Cr in groundwater. PMID- 11896261 TI - Paleontology. East of Eden at the Paleocene/Eocene boundary. PMID- 11896263 TI - Microbiology. A tail of two specifi-cities. PMID- 11896262 TI - Immunology. A perfect mismatch. PMID- 11896264 TI - Plant biology. Resisting attack. PMID- 11896266 TI - Improving the health of the global poor. AB - We analyzed the technical basis for a major global program to reduce disease among the poor. Effective interventions exist against the few diseases which most account for excess mortality among the poor. Achieving high coverage of effective interventions requires a well-functioning health system, as well as overcoming a set of financial and nonfinancial constraints. The annual incremental cost would be between $40 billion and $52 billion by 2015 in 83 low-income and sub-Saharan African countries. Such a program is feasible and would avoid millions of child, maternal, and adult deaths annually in poor countries. PMID- 11896267 TI - Resources required for global tuberculosis control. AB - We estimate that to achieve the World Health Organization's tuberculosis control targets, the 22 high-burden countries (HBCs) that collectively account for approximately 80% of the world's tuberculosis cases require about $1 billion per year during the period 2001 to 2005. A further $0.2 billion per year is needed for low and lower-middle income countries outside the 22 HBCs. There is a resource gap of up to around $300 million per year. Substantial progress in tuberculosis control could be achieved with increased investment that is large in the context of existing spending, but small in the wider context of global health expenditure. PMID- 11896268 TI - Erasing the world's slow stain: strategies to beat multidrug-resistant tuberculosis. AB - Multidrug-resistant tuberculosis (MDR) is perceived as a growing hazard to human health worldwide. Judgments about the true scale of the problem, and strategies for containing it, need to come from a balanced appraisal of the epidemiological evidence. We conclude in this review that MDR is, and will probably remain, a locally severe problem; that epidemics can be prevented by fully exploiting the potential of standard short-course chemotherapy (SCC) based on cheap and safe first-line drugs; and that best-practice SCC may even reduce the incidence of MDR where it has already become endemic. On the basis of the available, imperfect data, we recommend a three-part response to the threat of MDR: widespread implementation of SCC as the cornerstone of good tuberculosis control, improved resistance testing and surveillance, and the careful introduction of second-line drugs after a sound evaluation of cost, effectiveness, and feasibility. PMID- 11896269 TI - Responding to the challenge of communicable disease in Europe. AB - In the 1960s and 1970s, communicable disease seemed a minor threat, but since then the emergence of new infections and the reemergence of old diseases has provoked a renewed focus on European communicable disease surveillance and control. A "network approach" among European countries has been successful in detecting some international outbreaks, but management and funding aspects remain unresolved. Surveillance outside the European Union has faced new challenges as a result of economic and political change following the collapse of communism. Subsequently, innovative international surveillance schemes are currently being implemented in the countries of Central and Eastern Europe and the former Soviet Union. The challenge for surveillance in Europe is to ensure that it has the capacity to meet both the needs of today and the diseases of the future. PMID- 11896270 TI - Extensive fungal diversity in plant roots. PMID- 11896271 TI - Atom-resolved imaging of dynamic shape changes in supported copper nanocrystals. AB - In situ transmission electron microscopy is used to obtain atom-resolved images of copper nanocrystals on different supports. These are catalysts for methanol synthesis and hydrocarbon conversion processes for fuel cells. The nanocrystals undergo dynamic reversible shape changes in response to changes in the gaseous environment. For zinc oxide-supported samples, the changes are caused both by adsorbate-induced changes in surface energies and by changes in the interfacial energy. For copper nanocrystals supported on silica, the support has negligible influence on the structure. Nanoparticle dynamics must be included in the description of catalytic and other properties of nanomaterials. In situ microscopy offers possibilities for obtaining the relevant atomic-scale insight. PMID- 11896272 TI - Lateral hopping of molecules induced by excitation of internal vibration mode. AB - We demonstrate electron-stimulated migration for carbon monoxide (CO) molecules adsorbed on the Pd(110) surface, which is initiated by the excitation of a high frequency (HF) vibrational mode (C-O stretching mode) with inelastic tunneling electrons from the tip of scanning tunneling microscopy. The hopping phenomenon, however, cannot be detected for CO/Cu(110), even though the hopping barrier is lower than in the CO/Pd(110) case. A theoretical model, which is based on the anharmonic coupling between low-frequency modes (the hindered-translational mode related to the lateral hopping) and the HF mode combined with electron-hole pair excitation, can explain why the hopping of CO is observed on Pd(110) but not on Cu(110). PMID- 11896273 TI - Laser-induced selectivity for dimerization versus polymerization of butadiene under pressure. AB - The pressure-induced chemical reaction of liquid butadiene was studied by Fourier transform infrared spectroscopy in a diamond anvil cell. Dimerization was found to occur above 0.7 gigapascal, giving vinylcyclohexene according to a cyclo addiction reaction and only a trace amount of polybutadiene forms. By irradiating the high-pressure sample with a few milliwatts of the 488-nanometer argon+ laser line, the dimerization was completely inhibited, and the rapid formation of pure trans-polybutadiene was observed. The use of different excitation wavelength allows us to emphasize the selectivity of the process and to identify the active role of the 2(1)Ag state in this pressure- and laser-induced chemical reaction. PMID- 11896274 TI - Chromium isotopes and the fate of hexavalent chromium in the environment. AB - Measurements of chromium (Cr) stable-isotope fractionation in laboratory experiments and natural waters show that lighter isotopes reacted preferentially during Cr(VI) reduction by magnetite and sediments. The 53Cr/52Cr ratio of the product was 3.4 +/- 0.1 per mil less than that of the reactant. 53Cr/52Cr shifts in water samples indicate the extent of reduction, a critical process that renders toxic Cr(VI) in the environment immobile and less toxic. PMID- 11896275 TI - Mammalian dispersal at the Paleocene/Eocene boundary. AB - A profound faunal reorganization occurred near the Paleocene/Eocene boundary, when several groups of mammals abruptly appeared on the Holarctic continents. To test the hypothesis that this event featured the dispersal of groups from Asia to North America and Europe, we used isotope stratigraphy, magnetostratigraphy, and quantitative biochronology to constrain the relative age of important Asian faunas. The extinct family Hyaenodontidae appeared in Asia before it did so in North America, and the modern orders Primates, Artiodactyla, and Perissodactyla first appeared in Asia at or before the Paleocene/Eocene boundary. These results are consistent with Asia being a center for early mammalian origination. PMID- 11896276 TI - Natural iron isotope variations in human blood. AB - Isotopic analysis of human blood and liver and muscle tissue indicates that each individual bears a long-term iron (Fe) isotope signature in the blood. Blood and tissue differ slightly in isotopic composition and are depleted by up to 2.6 per mil in 56Fe relative to 54Fe when compared to dietary Fe. The 56Fe/54Fe isotope ratio in the blood of males is, on average, lower by 0.3 per mil than that of females. These results suggest that Fe isotope effects in the blood reflect differences in intestinal Fe absorption between individuals and genotypes. PMID- 11896277 TI - Metabolic activity of subsurface life in deep-sea sediments. AB - Global maps of sulfate and methane in marine sediments reveal two provinces of subsurface metabolic activity: a sulfate-rich open-ocean province, and an ocean margin province where sulfate is limited to shallow sediments. Methane is produced in both regions but is abundant only in sulfate-depleted sediments. Metabolic activity is greatest in narrow zones of sulfate-reducing methane oxidation along ocean margins. The metabolic rates of subseafloor life are orders of magnitude lower than those of life on Earth's surface. Most microorganisms in subseafloor sediments are either inactive or adapted for extraordinarily low metabolic activity. PMID- 11896278 TI - Demographic characteristics and population dynamical patterns of solitary birds. AB - In birds and many other animals, there are large interspecific differences in the magnitude of annual variation in population size. Using time-series data on populations of solitary bird species, we found that fluctuations in population size of solitary birds were affected by the deterministic characteristics of the population dynamics as well as the stochastic factors. In species with highly variable populations, annual variation in recruitment was positively related to the return rate of adults between successive breeding seasons. In stable populations, more recruits were found in years with low return rates of breeding adults. This identifies a gradient, associated with the position of the species along a "slow-fast" continuum of life history variation, from highly variable populations with a recruitment-driven demography to stable, strongly density regulated populations with a survival-restricted demography. These results suggest that patterns in avian population fluctuations can be predicted from a knowledge of life-history characteristics and/or temporal variation in certain demographic traits. PMID- 11896279 TI - Reverse transcriptase-mediated tropism switching in Bordetella bacteriophage. AB - Host-pathogen interactions are often driven by mechanisms that promote genetic variability. We have identified a group of temperate bacteriophages that generate diversity in a gene, designated mtd (major tropism determinant), which specifies tropism for receptor molecules on host Bordetella species. Tropism switching is the result of a template-dependent, reverse transcriptase-mediated process that introduces nucleotide substitutions at defined locations within mtd. This cassette-based mechanism is capable of providing a vast repertoire of potential ligand-receptor interactions. PMID- 11896280 TI - Influence of SHIP on the NK repertoire and allogeneic bone marrow transplantation. AB - Natural killer cell (NK) receptors for major histocompatibility complex (MHC) class I influence engraftment and graft-versus-tumor effects after allogeneic bone marrow transplantation. We find that SH2-containing inositol phosphatase (SHIP) influences the repertoire of NK receptors. In adult SHIP-/- mice, the NK compartment is dominated by cells that express two inhibitory receptors capable of binding either self or allogeneic MHC ligands. This promiscuous repertoire has significant functional consequences, because SHIP-/- mice fail to reject fully mismatched allogeneic marrow grafts and show enhanced survival after such transplants. Thus, SHIP plays an important role in two processes that limit the success of allogeneic marrow transplantation: graft rejection and graft-versus host disease. PMID- 11896281 TI - Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants. AB - T cells that accompany allogeneic hematopoietic grafts for treating leukemia enhance engraftment and mediate the graft-versus-leukemia effect. Unfortunately, alloreactive T cells also cause graft-versus-host disease (GVHD). T cell depletion prevents GVHD but increases the risk of graft rejection and leukemic relapse. In human transplants, we show that donor-versus-recipient natural killer (NK)-cell alloreactivity could eliminate leukemia relapse and graft rejection and protect patients against GVHD. In mice, the pretransplant infusion of alloreactive NK cells obviated the need for high-intensity conditioning and reduced GVHD. NK cell alloreactivity may thus provide a powerful tool for enhancing the efficacy and safety of allogeneic hematopoietic transplantation. PMID- 11896282 TI - Tech.Sight. Analyzing biomolecular interactions. PMID- 11896284 TI - Interaction of exposure concentration and duration in determining acute toxic effects of sarin vapor in rats. AB - Sarin (GB) vapor exposure is associated with both systemic and local toxic effects occurring primarily via the inhalation and ocular routes. The objective of these studies was to develop models for predicting dose-response effects of GB vapor concentrations as a function of exposure duration. Thus, the probability of GB vapor-induced lethality was estimated in rats exposed to various combinations of exposure concentration and duration. Groups of male and female Sprague-Dawley rats were exposed to one of a series of GB vapor concentrations for a single duration (5-360 min) in a whole-body dynamic chamber. The onset of clinical signs and changes in blood cholinesterase activity were measured with each exposure. Separate effective concentrations for lethality in 50% of the exposed population (LC50) and corresponding dose-response slopes were determined for each exposure duration by the Bliss probit method. Contrary to that predicted by Haber's rule, the interaction of LC50 x time (LCT50) values increased with exposure duration (i.e., the CT for 50% lethality in the exposed population and corresponding dose response slope was not constant over time). A plot of log (LCT50) versus log (exposure time) showed significant curvature. Predictive models derived from multifactor probit analysis of results describing the relationship between exposure conditions and probability of lethality in the rat are discussed. Overall, female rats were more sensitive to GB vapor toxicity than male rats over the range of exposure concentration and duration studied. Miosis was the initial clinical sign noted after the start of GB vapor exposure. Although blood cholinesterase activity was significantly inhibited by GB vapor exposure, poor correlation between cholinesterase inhibition and exposure conditions or cholinesterase inhibition and severity of clinical signs was noted. PMID- 11896285 TI - Evaluation of child/adult pharmacokinetic differences from a database derived from the therapeutic drug literature. AB - Pharmacokinetics (PK) of xenobiotics can differ widely between children and adults due to physiological differences and the immaturity of enzyme systems and clearance mechanisms. This makes extrapolation of adult dosimetry estimates to children uncertain, especially at early postnatal ages. While there is very little PK data for environmental toxicants in children, there is a wealth of such data for therapeutic drugs. Using published literature, a Children's PK Database has been compiled which compares PK parameters between children and adults for 45 drugs. This has enabled comparison of child and adult PK function across a number of cytochrome P450 (CYP) pathways, as well as certain Phase II conjugation reactions and renal elimination. These comparisons indicate that premature and full-term neonates tend to have 3 to 9 times longer half-life than adults for the drugs included in the database. This difference disappears by 2-6 months of age. Beyond this age, half-life can be shorter than in adults for specific drugs and pathways. The range of neonate/adult half-life ratios exceeds the 3.16-fold factor commonly ascribed to interindividual PK variability. Thus, this uncertainty factor may not be adequate for certain chemicals in the early postnatal period. The current findings present a PK developmental profile that is relevant to environmental toxicants metabolized and cleared by the pathways represented in the current database. The manner in which this PK information can be applied to the risk assessment of children includes several different approaches: qualitative (e.g., enhanced discussion of uncertainties), semiquantitative (age group-specific adjustment factors), and quantitative (estimation of internal dosimetry in children via physiologically based PK modeling). PMID- 11896286 TI - Chloroform inhalation exposure conditions necessary to initiate liver toxicity in female B6C3F1 mice. AB - Chloroform is a nongenotoxic-cytotoxic carcinogen in rodent liver and kidney, including the female B6C3F1 mouse liver. Because tumors are secondary to events associated with cytolethality and regenerative cell proliferation, these end points are valid surrogates for tumor formation in cancer risk assessments. The purpose of the experiments presented here was to more clearly define the combinations of atmospheric concentration and duration of exposure necessary to induce cytolethality and regenerative cell proliferation in the sensitive female B6C3F1 mouse liver. Female B6C3F1 mice were exposed to chloroform by inhalation for 7 consecutive days using atmospheres of 10, 30, or 90 ppm and selected exposure times of 2, 6, 12, or 18 h/day. Bromodeoxyuridine (BrdU) was given the last 3.5 days via an implanted osmotic pump to label cells in S-phase. Labeled hepatocytes were visualized immunohistochemically, and the labeling index (LI) was determined as the percentage of cells in S-phase. LI was a more sensitive indicator of cellular damage than histopathological examination and is the more conservative end point for use in risk assessments. Significant concentration and exposure time related increases in LI were observed at 30 and 90 ppm but not at any 10-ppm exposure. These data defined an empirical relationship for the combinations of airborne exposure concentration and duration needed to induce cytolethality. These results suggest that concentrations of about 10 ppm or below will not induce hepatotoxicity in these mice regardless of exposure duration. Thus, the rate of production of toxic metabolites and the subsequent rate of cellular damage produced by a continual exposure of approximately 10 ppm chloroform are less than the maximum rates at which hepatocytes can detoxify those metabolites and repair any induced cellular damage. A physiologically based pharmacokinetic (PBPK) dosimetry model was used to compare anticipated responses in mice and humans and predicted that chloroform concentrations of approximately an order of magnitude greater than 10 ppm would be required to induce human liver toxicity. Thus, no safety factor to account for species to species extrapolation should be required in formulating a chloroform inhalation cancer risk assessment based on the dose x time inhalation data presented here. PMID- 11896287 TI - p53 heterozygosity alters the mRNA expression of p53 target genes in the bone marrow in response to inhaled benzene. AB - C57BL/6 Trp53 heterozygous (N5) mice (p53+/- mice) show an increased sensitivity to tumorigenesis following exposure to genotoxic compounds and are being used as an alternate animal model for carcinogenicity testing. However, there is relatively little data regarding the effect of p53 heterozygosity on the genomic and cellular responses of target tissues in these mice to toxic insult, especially under chronic exposure conditions used in carcinogenicity bioassays. We hypothesized that heterozygosity at the p53 locus in p53+/- mice alters the expression of bone marrow p53-regulated genes involved in cell cycle control and apoptosis during chronic genotoxic stress. We used real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) to examine gene expression alterations in bone marrow cells from C57BL/6 p53+/+ and isogenic p53+/- mice chronically exposed for 15 weeks to genotoxic and carcinogenic levels (100 ppm) of inhaled benzene. Examination of mRNA levels of p53-regulated genes involved in cell cycle control (p21, gadd45, and cyclin G) or apoptosis (bax and bcl-2) showed that during chronic genotoxic stress, bone marrow cells from p53+/+ mice expressed significantly higher levels of a majority of these genes compared to p53+/- bone marrow cells. Our results indicate that p53 heterozygosity results in a haploinsufficient phenotype in p53+/- bone marrow cells as evident by significantly altered mRNA levels of key genes involved in the p53-regulated DNA damage response pathway. PMID- 11896288 TI - Modulation of mammary gland development in prepubertal male rats exposed to genistein and methoxychlor. AB - The estrogenic isoflavone genistein is a common dietary component that has been shown to affect reproductive development in experimental animals at high doses. The objective of the present study was to examine interactions of genistein and the hormonally active pesticide methoxychlor on mammary gland development in juvenile rats. Timed-pregnant Sprague-Dawley rats were fed a soy- and alfalfa free diet containing different combinations of genistein (300 and 800 ppm) and methoxychlor (800 ppm). Rats were fed these diets starting on gestation day (GD)1 and continuing through pregnancy and lactation until postnatal day (PND) 22, when the pups were killed. Inguinal mammary glands from both female and male pups were processed as whole-mount preparations for morphometric analysis. The total glandular area and the numbers of branch points, lateral buds, and terminal end buds in the male rats were found to be significantly greater in the groups exposed to methoxychlor than those exposed to genistein only. These effects were not observed in the female rats. In the male rats, methoxychlor had the most prominent effect on elongating the glandular ducts, while genistein enhanced the ductile branching. The 2 compounds in combination promoted the development of alveolar-lobular structure, an effect not observed with either compound alone. Immunostaining for proliferating cell nuclear antigen revealed a high percentage of immunopositive cells in the mammary epithelia of the males exposed to methoxychlor and genistein (800 ppm) compared to the controls. While no significant changes in serum levels of mammotrophic hormones were detected, increased immunostaining for insulin-like growth factor-1 receptor, estrogen receptor alpha, and progesterone receptor in the genistein + methoxychlor group suggested that local factors involved in regulating mammary growth may have played a role in propagating the endocrine effects of these two compounds. These results indicated that the mammary glands of juvenile male rather than juvenile female rats may be more sensitive to certain endocrine-active compounds and that high levels of phytoestrogens have the potential to alter the toxicological behaviors of other hormone mimics. PMID- 11896289 TI - Ranking of allergenic potency of rubber chemicals in a modified local lymph node assay. AB - A modified local lymph node assay (LLNA) with ex vivo tritium thymidine (3H-TdR) labeling of the proliferating lymph node cells was used for determination of the allergenic potency of chemicals used in the production of rubber for latex medical gloves. Fifteen chemicals known to induce contact hypersensitivity reactions in man, including various thiuram, carbamate, and benzothiazole compounds, and one amine were tested. The EC3 (effective concentration inducing a 3-fold increase in proliferation of lymph node cells [Stimulation Index, SI = 3]) was calculated with nonlinear regression analysis, including a bootstrap method for determination of the 5-95% confidence interval of the EC3 value. This procedure identified 14 out of the 15 chemicals tested as sensitizers, while for one chemical, ZDBC, no EC3 could be calculated due to low responses and a lack of a dose-response relationship in the data obtained. The ranking order of the chemicals with increasing EC3 values (and thus decreasing allergenic potency) was found to be in the following order: ZDEC < TMTD < TETD < ZPC < ZDMC < MBTS < PTD < TMTM < MBT < MBI < PTT < ZMBT < TBTD < DEA < ZDBC. Our results indicate that the chemicals of choice for use in the production of natural rubber latex products would be for the thiuram compounds, TBTD; for the carbamates, ZDBC; and for the benzothiazoles, ZMBT. However, one has to be aware that besides potency, the total amount of residual chemical present in the final product is also important for allergy induction. PMID- 11896290 TI - Acetaminophen-induced proliferation of estrogen-responsive breast cancer cells is associated with increases in c-myc RNA expression and NF-kappaB activity. AB - Studies reported here tested the hypothesis that acetaminophen stimulates proliferation of E2-responsive cells by inducing expression of E2-regulated genes. Ribonuclease protection assays compared the effects of acetaminophen and E2 on expression of selected genes (c-myc, c-fos, cyclin D1, bcl-2, bax, gadd45, mcl-1, p53, p21(CIP1/WAF1), and bcl-xL) in E2-responsive breast cancer (MCF-7) and endometrial adenocarcinoma (Ishikawa) cells as well as in E2-nonresponsive (MDA-MB-231) breast cancer cells. Acetaminophen and E2 increased c-myc RNA levels in MCF-7 cells, consistent with a mitogenic activity of these compounds in MCF-7 cells. However, the magnitude and time course of acetaminophen and E2 induction of c-myc differed. Neither acetaminophen nor E2 induced c-myc in MDA-MB-231 cells, whereas E2, but not acetaminophen, weakly induced c-myc expression in Ishikawa cells. Furthermore, in these 3 cell types, the expression patterns of the other genes differed dramatically in response to acetaminophen and to E2, indicating that acetaminophen does not activate ER as a transcription factor in the same manner as does E2. Additionally, gel shift assays demonstrated that in MCF-7 cells, acetaminophen increased NF-kappaB activity approximately 40% and did not alter AP-1 activity, whereas E2 increased AP-1 activity approximately 50% and did not increase NF-B activity. These studies indicate that acetaminophen effects on gene expression and cell proliferation depend more on cell type/context than on the presence of ER. PMID- 11896291 TI - Structural determinants of fluorochemical-induced mitochondrial dysfunction. AB - Perfluorooctanoate (PFOA) and perfluorooctanesulfonate (PFOS) are thought to induce peroxisome proliferation and interfere with mitochondrial metabolic pathways. Direct measurements revealed that PFOA and the unsubstituted sulfonamide of perfluorooctane (FOSA) uncouple mitochondrial respiration by increasing proton conductance. The purpose of this investigation was to characterize structural determinants responsible for the mitochondrial uncoupling effect of several structurally related fluorochemicals. Included in the study were PFOA, PFOS, FOSA, the N-acetate of FOSA (perfluorooctanesulfonamidoacetate, FOSAA), N-ethylperfluorooctanesulfonamide (N-EtFOSA), and the N-ethyl alcohol [2 (N-ethylperfluorooctanesulfonamido)ethyl alcohol, N-EtFOSE] and N-acetic acid (N ethylperfluorooctanesulfonamidoacetate, N-EtFOSAA) of N-EtFOSA. Each test compound was dissolved in ethanol and added directly to an incubation medium containing substrate-energized rat liver mitochondria. Mitochondrial respiration and membrane potential were measured concurrently using an oxygen electrode and a TPP+ -selective electrode, respectively. All of the compounds tested, at sufficiently high concentrations, had the capacity to interfere with mitochondrial respiration, albeit via different mechanisms and with varying potencies. At sufficiently high concentrations, the free acids PFOA and PFOS caused a slight increase in the intrinsic proton leak of the mitochondrial inner membrane, which resembled a surfactant-like change in membrane fluidity. Similar effects were observed with the sulfonamide N-EtFOSE. Another fully substituted sulfonamide, N-EtFOSAA, at high concentrations caused inhibition of respiration, the release of cytochrome c, and high-amplitude swelling of mitochondria. The swelling was prevented by cyclosporin A or by EGTA, indicating that this compound induced the mitochondrial permeability transition. The unsubstituted and mono substituted amides FOSA, N-EtFOSA, and FOSAA all exerted a strong uncoupling effect on mitochondria resembling that of protonophoric uncouplers. Among these compounds, FOSA was a very potent uncoupler of oxidative phosphorylation, with an IC50 of approximately 1 microM. These data suggest that the protonated nitrogen atom with a favorable pKa is essential for the uncoupling action of perfluorooctane sulfonamides in mitochondria, which may be critical to the mechanism by which these compounds interfere with mitochondrial metabolism to induce peroxisome proliferation in vivo. PMID- 11896292 TI - Aluminum chloride induces retinal changes in the rat. AB - We studied rat retinal changes due to aluminum (Al) toxicosis with a transmission electron microscope (TEM) and an energy dispersive X-ray analyzer (EDXA). Normal 4-week-old Wistar Kyoto rats were divided randomly into Al toxicosis and control groups. The Al toxicosis group was injected ip with 0.3 ml of 4% aluminum chloride (AlCl3) per day every day for 16 weeks. The retina was examined with a TEM and EDXA at 8, 12, and 16 weeks after starting injections with AlCl3. There was a statistically significant increase in the serum Al concentration in the Al toxicosis group (p < 0.001). We observed prominent pathologic changes at 16 weeks after the first injections. Thin retinal pigment epithelium (RPE), and disappearance of the photoreceptor outer and inner segments and nuclei were observed. There were high-density irregular granules in the outer and inner plexiform layers and in the inner nuclear layer. We found dense granules in the cells, which remained between the RPE and the inner nuclear layer. EDXA detected Al in the high-density irregular granules in these areas. Al injected ip caused accumulation of Al in the rat retina and the destruction of photoreceptor cells. These findings indicate that Al is toxic to the retina. PMID- 11896293 TI - Disposition of inhaled mercury vapor in pregnant rats: maternal toxicity and effects on developmental outcome. AB - The disposition and toxicity of inhaled elemental mercury (Hg0) vapor for pregnant Long-Evans rats, and potential adverse effects on reproductive outcome were investigated. Rats were exposed to 0, 1, 2, 4, or 8 mg Hg0/m(3) for 2 h/day from gestation day (GD) 6 through GD 15. Maternal toxicity occurred primarily in rats exposed to 4 and 8 mg/m(3) and was manifested as a concentration-related decrease in body weight gain and mild nephrotoxicity. Control rats gained about 13% of their initial body weight during the 10-day exposure. Rats exposed to 4 mg/m(3) Hg0 gained about 7% less than controls, and rats exposed to 8 mg/m(3) Hg0 lost about 17% of their initial body weight during the 10-day exposure period. Maternal kidney weights were significantly increased in the 4 and 8 mg/m(3) concentration groups, and urinalysis revealed increased levels of protein and alkaline phosphatase activity in urine of all Hg0-exposed rats. Dams exposed to 8 mg/m(3) were euthanized in moribund condition on postnatal day (PND) 1. There was no histopathological evidence of toxicity in maternal lung, liver, or kidney of exposed rats at GD 6, GD 15, or PND 1. The incidence of resorptions was significantly increased, litter size and PND 1 neonatal body weights were significantly decreased only in the 8-mg/m(3) group. Total Hg concentrations in maternal tissues increased with increasing number of exposure days and concentration. In general, approximately 70% of Hg was eliminated from maternal tissues during the week following the last exposure (GD 15 to PND 1). Elimination of Hg from maternal brain and kidney was slower than in other tissues, possibly due to higher levels of metallothionein. Total Hg concentrations in fetal tissues increased with increasing number of exposure days and concentration, demonstrating that a significant amount of Hg crossed the placenta. One week after the last exposure, significant amounts of Hg were still present in brain, liver, and kidney of PND 1 neonates. Metallothionein levels in neonatal tissues were not significantly increased by exposure to 4 mg/m(3) Hg0. The total amount of Hg in neonatal brain (ng/brain) continued to increase after termination of inhalation exposure, suggesting a redistribution of Hg from the dam to neonatal brain. These data demonstrate that inhaled Hg0 vapor is distributed to all maternal and fetal tissues in a dose-dependent manner. Adverse effects of Hg on developmental outcome occurred only at a concentration that caused maternal toxicity. PMID- 11896294 TI - Prevention of vitamin A teratogenesis by phytol or phytanic acid results from reduced metabolism of retinol to the teratogenic metabolite, all-trans-retinoic acid. AB - Previous studies in our laboratory showed a synergistic interaction of synthetic ligands selective for the retinoid receptors RAR and RXR in regard to teratogenic effects produced in mice (M. M. Elmazar et al., 2001, TOXICOL: Appl. Pharmacol. 170, 2-9). In the present study the influence of phytol and phytanic acid (a RXR selective ligand) on the teratogenicity of retinol and the RAR-selective ligand all-trans-retinoic acid was investigated by coadministration experiments on day 8.25 of gestation in NMRI mice. Phytol and phytanic acid, noneffective when administered alone, did not potentiate the teratogenicity induced by retinol or all-trans-retinoic acid. On the contrary, phytol and phytanic acid greatly reduced retinol-induced teratogenic effects (ear anotia, tail defects, exencephaly). The effect of phytol on all-trans-retinoic acid teratogenesis was limited (only resorptions and tail defects were reduced). Pharmacokinetic studies in nonpregnant animals revealed that phytol coadministration with retinol reduced plasma levels of retinol and retinyl esters, and drastically reduced the levels of the teratogenic retinol metabolite, all-trans-retinoic acid. Phytanic acid also reduced the oxidative metabolism and teratogenic effects of retinol. These results indicate that phytol and phytanic acid did not synergize with retinol and all-trans-retinoic acid in our mouse teratogenesis model. Instead, phytol and phytanic acid effectively blocked the teratogenic effects of retinol by drastically reducing the metabolic production of all-trans-retinoic acid. Phytol and phytanic acid may be useful for the prevention of vitamin A teratogenicity. PMID- 11896296 TI - Effect of diesel exhaust particle extracts on induction of oral tolerance in mice. AB - We examined the effect of diesel exhaust particle (DEP) extracts on oral tolerance in mice. For this examination, a single DEP sample was consecutively extracted with hexane (HEX-DEP), benzene (BEN-DEP), dichloromethane (DIC-DEP), methanol (MET-DEP), and 1 M ammonia (AMM-DEP). Residues unextracted (UNE-DEP) with the last extraction solvent 1 M ammonia were also used to test their ability to induce oral tolerance. To immunize mice, hen egg lysozyme (HEL) emulsified with an equal volume of CFA was injected sc (day 0). Oral tolerance was induced by feeding 10 mg HEL on days -5, -4, -3, -2, and -1. DEP, each DEP extract, and UNE-DEP were intranasally administered immediately after each feeding of HEL. The results showed that oral administration of HEL markedly suppressed production of anti-HEL IgG antibodies as well as proliferative responses of spleen cells to HEL. The suppression of anti-HEL IgG antibody production and the cell proliferation by the oral antigen was significantly blocked by DEP, DIC-, AMM-, and UNE-DEP. Neither HEX-, BEN-, nor MET-DEP modulated the orally induced suppression of these immune responses. When the levels of anti-HEL IgG2a antibodies and IFN-gamma (Th1 responses) and anti-HEL IgG1 antibodies and IL-4 (Th2 responses) were determined, DEP and DIC-DEP diminished the suppression of both Th1 and Th2 responses observed following oral administration of HEL. In contrast, UNE- and AMM-DEP prevented the reduction of Th1 but not Th2, and Th2 but not Th1 oral tolerance, respectively. Thus, UNE-DEP appears to contain compounds that block induction of Th1 but not Th2 oral tolerance, whereas AMM-DEP have compounds that abrogate induction of Th2 but not Th1 oral tolerance. DIC DEP, as well as DEP, appear to contain components that block induction of both Th1 and Th2 oral tolerance. As oral tolerance is thought to play a critical role in preventing Th1 as well as Th2 food allergy, the blockade of oral tolerance by these DEP extracts suggests that DEP may contain compounds different in hydrophobicity associated with the cause of such adverse immunologic responses to food proteins. PMID- 11896295 TI - Developmental stage-specific effects of perinatal 2,3,7,8-tetrachlorodibenzo-p dioxin exposure on reproductive organs of male rat offspring. AB - Exposure to a relatively low dose of 2,3,7,8-tetrachlorodebenzo-p-dioxin (TCDD) during mid-gestation induces a reduction of ventral prostate weight in rat offspring. Recently we reported that a single administration of TCDD (12.5-800 ng/kg body weight) to pregnant Holtzman rats on gestational day (GD) 15 caused a decrease in androgen receptor (AR) mRNA level in the ventral prostate during the prepubertal period, and we proposed that this reduction of AR mRNA is one of the most sensitive adverse endpoints due to perinatal exposure to TCDD (S. Ohsako et al., 2001, TOXICOL: Sci. 60, 132-143). In the present study, to investigate the mechanism of a decrease in AR mRNA level, we administered TCDD to rats at other developmental stages and compared possible alterations of the male reproductive system. Pregnant Sprague-Dawley rats were given a single oral dose of 1 microg TCDD/kg body weight on GD 15 or GD 18, or male pups born from untreated dams were subcutaneously given a single dose of 1 microg TCDD/kg body weight on postnatal day 2 (PND 2). Offspring exposed on GD 15, GD 18, and PND 2 were sacrificed on PND 70. TCDD exposure on GD 15 resulted in significant decreases in the urogenital complex and ventral prostate weights and urogenital-glans penis length of male rat offspring, but not on GD 18 and PND 2. Testicular and epididymal weights were also lower than control group only in the TCDD-exposed GD 15 group. Anogenital distance was significantly reduced in the TCDD-exposed GD 15 and GD 18 groups, but not in the TCDD-exposed PND 2 group. Semiquantitative RT-PCR analysis showed that AR mRNA levels were decreased in the TCDD-exposed GD 15 group only, and that the constitutive level of cytochrome P450 1A1 (CYP1A1) mRNA in the ventral prostate was not changed by TCDD in any of the exposed groups. No changes in AR mRNA level were detected in the testis or brain in any of the TCDD-exposed groups. These results suggest the presence of a critical window during development with regard to impairments of male reproductive organs by in utero and lactational exposure to a low dose of TCDD. PMID- 11896297 TI - Dose-response modeling of continuous endpoints. AB - A family of (nested) dose-response models is introduced herein that can be used for describing the change in any continuous endpoint as a function of dose. A member from this family of models may be selected using the likelihood ratio test as a criterion, to prevent overparameterization. The proposed methodology provides for a formal approach of model selection, and a transparent way of assessing the benchmark dose. Apart from a number of natural constraints, the model expressions follow from an obvious way of quantifying differences in sensitivity between populations. As a consequence, dose-response data that relate to both sexes can be efficiently analyzed by incorporating the data from both sexes in the same analysis, even if the sexes are not equally sensitive to the compound studied. The idea of differences in sensitivity is closely related to the assessment factors used in risk assessment. Thus, the models are directly applicable to estimating such factors, if data concerning populations to be compared are available. Such information is valuable for further validation or adjustment of default assessment factors, as well as for informing distributional assessment factors in a probabilistic risk assessment. The various applications of the proposed methodology are illustrated by real data sets. PMID- 11896298 TI - Analysis of rodent growth data in toxicology studies. AB - To evaluate compound-related effects on the growth of rodents, body weight and food consumption data are commonly collected either weekly or biweekly in toxicology studies. Body weight gain, food consumption relative to body weight, and efficiency of food utilization can be derived from body weight and food consumption for each animal in an attempt to better understand the compound related effects. These five parameters are commonly analyzed in toxicology studies for each sex using a one-factor analysis of variance (ANOVA) at each collection point. The objective of this manuscript is to present an alternative approach to the evaluation of compound-related effects on body weight and food consumption data from both subchronic and chronic rodent toxicology studies. This approach is to perform a repeated-measures ANOVA on a selected set of parameters and analysis intervals. Compared with a standard one-factor ANOVA, this approach uses a statistical analysis method that has greater power and reduces the number of false-positive claims, and consequently provides a succinct yet comprehensive summary of the compound-related effects. Data from a mouse carcinogenicity study are included to illustrate this repeated-measures ANOVA approach to analyzing growth data in contrast with the one-factor ANOVA approach. PMID- 11896299 TI - Vinyl acetate-induced intracellular acidification: implications for risk assessment. AB - Cancerbioassays have demonstrated the carcinogenic activity of vinyl acetate in rodents. Tumors appear only at the site of contact and mechanistic data suggest that the carcinogenic mechanism involves carboxylesterase-mediated metabolism of vinyl acetate to acetic acid. It has been hypothesized that intracellular formation of acetate causes a reduction of intracellular pH (pH(i)) at noncytotoxic levels, but that prolonged exposure to reduced pH(i) is cytotoxic and/or mitogenic and drives proliferative responses. Coupled with exposure to metabolically formed acetaldehyde at high administered concentrations, nonlinear dose-response curves for epithelial tumors are produced. Freshly isolated rat hepatocytes were used as a model system to test the concept that exposure of cells to vinyl acetate causes a reduction in pH(i). Quantitative fluorescence imaging ratio microscopy showed that exposure of hepatocytes to vinyl acetate concentrations ranging from 10 to 1000 microM caused rapid and sustained reductions of approximately 0.03 to 0.65 pH units. Cellular acidification was rapidly reversed to control pH(i) upon removal of vinyl acetate. There was minimal accumulation of protons during the exposure period, as suggested by minor differences in pH(i) of cells with or without prior exposure to vinyl acetate. The effect of vinyl acetate on pH(i) was attenuated by prior exposure to the carboxylesterase inhibitor bis(p-nitrophenyl)phosphate. These results support the concept that intracellular acidification is a sentinel pharmacodynamic response of cells to vinyl acetate exposure and that pH(i) is an appropriate metric dose for use in quantitative risk assessments of cancer and noncancer human health risk assessment. PMID- 11896300 TI - Electrocardiographic changes during exposure to residual oil fly ash (ROFA) particles in a rat model of myocardial infarction. AB - Epidemiological studies have reported a positive association of short-term increases in ambient particulate matter (PM) with daily mortality and hospital admissions for cardiovascular disease. Although patients with cardiopulmonary disease appear to be most at risk, particulate-related cardiac effects following myocardial infarction (MI) have not been examined. To improve understanding of mechanisms, we developed and tested a model for investigating the effects of inhaled PM on arrhythmias and heart rate variability (HRV), a measure of autonomic nervous system activity, in rats with acute MI. Left-ventricular MI was induced in 31 Sprague-Dawley rats by thermocoagulation of the left coronary artery; 32 additional rats served as sham-operated controls. Diazepam-sedated rats were exposed (1 h) to residual oil fly ash (ROFA), carbon black, or room air at 12-18 h after surgery. Each exposure was immediately preceded and followed by a 1-h exposure to room air (baseline and recovery periods, respectively). Lead-II electrocardiograms were recorded. In the MI group, 41% of rats exhibited one or more premature ventricular complexes (PVCs) during the baseline period. Exposure to ROFA, but not to carbon black or room air, increased arrhythmia frequency in animals with preexisting PVCs. Furthermore, MI rats exposed to ROFA, but not to carbon black or room air, decreased HRV. There was no difference in arrhythmia frequency or HRV among sham-operated animals. These results underscore the usefulness of this model for elucidating the physiologic mechanisms of pollution induced cardiovascular arrhythmias and contribute to defining the specific constituents of ambient particles responsible for arrhythmias. PMID- 11896301 TI - Prediction of skin irritation from organic chemicals using membrane-interaction QSAR analysis. AB - Membrane-interaction (MI) quantitative structure activity relationship (QSAR) analysis was carried out for a training set of 22 hydroxy organic compounds for which the Draize skin irritation scores, PII, had been determined. Significant MI QSAR models were constructed in which skin irritation potency is predicted to increase with (1) increasing effective concentration of the compound available for uptake into phospholipid-rich regions of a cellular membrane, (2) increasing binding of the compound to the phospholipid-rich regions of a cellular membrane, and (3) the chemical reactivity of the compound as reflected by the highest occupied molecular orbital (HOMO) and/or lowest unoccupied molecular orbital (LUMO) of the molecule. Overall, the MI-QSAR models constructed for skin irritation are very similar, with respect to the types of descriptors, to those found for eye irritation. In turn, the skin irritation MI-QSAR models suggest a similar molecular mechanism of action to that postulated for eye irritation from MI-QSAR analysis. Significant and predictive QSAR models cannot be constructed unless test compound-membrane interaction descriptors are computed and used to build the QSAR models. PMID- 11896302 TI - Acute and subchronic mammalian toxicity of naphthenic acids from oil sands tailings. AB - Naphthenic acids are the most significant environmental contaminants resulting from petroleum extraction from oil sands deposits. In this study, a mixture of naphthenic acids isolated from Athabasca oil sands (AOS) tailings pond water was used in acute and subchronic toxicity tests with rodents, in order to assess potential risks posed to terrestrial wildlife. Dosages were chosen to bracket worst-case environmental exposure scenarios. In acute tests, adult female Wistar rats were given single po dosages of naphthenic acids at either 3, 30, or 300 mg per kg body weight (mg/kg), while adult male rats received 300 mg/kg. Food consumption was temporarily suppressed in the high-dose groups of both sexes. Following euthanasia 14 days later, histopathology revealed a significant incidence of pericholangitis in the high-dose group of both sexes, suggesting hepatotoxicity as an acute effect. Other histological lesions included brain hemorrhage in high-dose males, and cardiac periarteriolar necrosis and fibrosis in female rats. In subchronic tests, naphthenic acids were po administered to female Wistar rats at 0.6, 6, or 60 mg/kg, 5 days per week for 90 days. Results again suggested the liver as a potential target organ. The relative liver weight in the high-dose group was 35% higher than in controls. Biochemical analysis revealed elevated blood amylase (30% above controls) and hypocholesterolemia (43% below controls) in high-dose rats. Excessive hepatic glycogen accumulation was observed in 42% of animals in this group. These results indicate that, under worst-case exposure conditions, acute toxicity is unlikely in wild mammals exposed to naphthenic acids in AOS tailings pond water, but repeated exposure may have adverse health effects. PMID- 11896303 TI - Carcinogenicity bioassay of bisphenol A. PMID- 11896304 TI - Transfusion medicine illustrated. Giant pronormoblasts due to parvovirus B19 infection. PMID- 11896306 TI - Analysis of factors associated with low peripheral blood progenitor cell collection in normal donors. AB - BACKGROUND: Predictive factors of the response to rHuG-CSF in normal donors have not been extensively studied. STUDY DESIGN AND METHODS: We analyzed factors influencing CD34+ cell yield in the 1st day of collection in 261 healthy donors from the Spanish National Donor Registry. The median age was 38 years (range, 2 72). The median dose of rHuG-CSF was 10 microg per kg per day (range, 5-20) over 4 days. In 103 donors (40%), <4 x 10(6) per kg CD34+ cells were collected. The variables that were analyzed included age, sex, weight, basal complete blood cell count, dose, type of rHuGCSF and schedule of administration, and maximum WBC count before apheresis. RESULTS: By univariate analysis, the maximum WBC count (<50 vs. >or=50 x 10(9)/L, p = 0.004), advanced age (p = 0.008), and number of daily rHuG-CSF doses (one vs. two; p = 0.01) correlated with the number of CD34+ cells collected. By multivariate analysis, donors age (<38 vs. >or=38 years; p = 0.014) and a single daily dose of rHuG-CSF (p = 0.005) were the two variables that significantly predicted a low CD34+ cell yield. CONCLUSION: Donors' age, with a threshold of 38 years or more, and the rHuG-CSF schedule are the factors that significantly affected CD34+ cell mobilization and collection in healthy donors. PMID- 11896307 TI - Processing of peripheral blood progenitor cell components in improved clean areas does not reduce the rate of microbial contamination. AB - BACKGROUND: Microbial contamination of peripheral blood progenitor cell components (PBPCs) may cause severe complications in immunosuppressed recipients. Therefore, principles of Good Manufacturing Practice (GMP) are applicable for processing of PBPC components to reduce potential risks of contamination. STUDY DESIGN AND METHODS: It was investigated in a retrospective study whether the microbial contamination of PBPC components could be reduced after processing in improved clean areas according to the "Manufacture of Sterile Medicinal Products." Starting in 1994, a total of 1478 autologous and allogeneic PBPC components have been collected and processed into 3149 cryopreservation bags at the Department of Transfusion Medicine. Sterility testing was performed for all bags. Until December 1998, 783 PBPC components were processed at a clean bench only (group I). Thereafter, 695 PBPC components have been processed at a clean bench located in a clean area with an airlock system for personnel and equipment (group II). RESULTS: In group I, 16 of 1555 bags (1.03%) showed positive results in the first sterility testing. In group II, 21 of 1594 bags (1.32%) were positive (p = NS). The clinical follow-up was inconspicuous. CONCLUSION: Microbial contamination of PBPC components could not be reduced by installation of improved clean area conditions. PMID- 11896308 TI - Acquired FV inhibitors: a needless iatrogenic complication of bovine thrombin exposure. AB - BACKGROUND: FV inhibitors are a largely preventable iatrogenic coagulopathy in which the frequency is increasing in clinical practice. STUDY DESIGN AND METHODS: Three cases associated with our institution are reported. A systematic review of the MEDLINE database was performed, and reference lists were reviewed to identify relevant publications. RESULTS: One hundred twenty-six cases of FV inhibitors have been reported in the world's literature. Eighty-seven have been reported in the last decade, of which two thirds are due to exposure to bovine thrombin. Bovine thrombin-associated FV antibodies develop in 40 to 66 percent of cardiac surgery patients and in 20 percent of neurosurgery patients. Thirty-three percent of reported patients developed bleeding complications. Inhibitors persisted on average 2.3 months. Standard coagulation assays do not reliably predict clinical manifestations. Multimodality therapy, including immunosuppression, is useful for treatment of symptomatic patients. CONCLUSIONS: FV inhibitors are a common complication of bovine thrombin exposure that can have devastating clinical consequences. Transfusion medicine specialists and hematologists can play a critical role in reducing the incidence of FV inhibitors by educating the medical community about safer alternative fibrin sealants. PMID- 11896309 TI - A randomized, blinded trial comparing the hemostatic effects of pentastarch versus hetastarch. AB - BACKGROUND: HES solutions provide a sterile, alternative colloidal fluid to albumin solutions and/or plasma in the management of patients who need plasma volume expansion. Solutions of HES are widely accepted internationally but are used only modestly in the United States, largely because of concerns over hemostasis. STUDY DESIGN AND METHODS: A randomized, blinded, two-arm trial comparing the hemostatic effects of pentastarch versus hetastarch when infused in the clinically relevant dose of 90 g of HES dissolved in 1.5 L of saline was conducted. Multiple studies of fibrin clot formation, fibrinogen/fibrinolysis, and platelet (PLT) functions were performed before and on multiple occasions for 70 days following HES infusion. RESULTS: Several significant abnormalities of hemostasis assay results occurred following HES infusions, with hetastarch causing significantly greater abnormalities than pentastarch. Individual clotting proteins and blood PLTs fell modestly because of plasma volume expansion and hemodilution. A fall in excess of that caused by hemodilution was demonstrated for von Willebrand factor antigen plus its associated FVIII and ristocetin cofactor activities. The partial thromboplastin time was prolonged, whereas the thrombin time was shortened. Plt function abnormalities were seen in most subjects to a modest degree. Studies of fibrinolysis were normal. CONCLUSIONS: Solutions of hetastarch produce significant abnormalities of some hemostasis laboratory results when infused at clinically relevant doses, but it is unlikely that the modest hemostatic abnormalities produced at these doses per se would lead to clinical bleeding. Hetastarch causes greater hemostatic abnormalities than pentastarch, and because both HES solutions have comparable plasma volume expanding effects, it is reasonable to prefer pentastarch as a plasma volume expander. PMID- 11896311 TI - In utero development of a warm-reactive autoantibody in a severely jaundiced neonate. AB - BACKGROUND: The fetus and neonate are widely considered to be immunologically immature. However, there are rare case reports of RBC alloantibody and autoantibody development. STUDY DESIGN AND METHODS: This report describes the case of a severely jaundiced full-term boy neonate presenting at birth with an IgG warm-reactive autoantibody. RESULTS: Mother and neonate were both blood group A, D+. The mother had a negative antibody screen at 18 weeks' gestation and a negative DAT and antibody screen at the time of delivery. The neonate was born with a strongly reactive DAT (IgG) and a panreactive eluate. The serum also contained a panreactive antibody, and all crossmatches were incompatible. The neonate had a bilirubin of 12.5 mg per dL at birth, which peaked at 22.5 mg per dL. However, there was no overt evidence of hemolysis, as evidenced by normal serial Hct levels and reticulocyte counts. The neonate responded well to phototherapy and did not require either simple or exchange transfusion. The neonate's warm-reactive autoantibody maintained its original strength of reactivity on follow-up testing performed at 2 weeks and 2 months of age. CONCLUSIONS: This report describes a rare case of apparent in utero RBC autoantibody development. The fetal/neonatal immune response to blood group antigens is reviewed. PMID- 11896310 TI - Clinical significance of RBC alloantibodies and autoantibodies in sickle cell patients who received transfusions. AB - BACKGROUND: The clinical significance of alloimmunization to RBC antigens in sickle cell patients was analyzed by a retrospective review of the records of pediatric and adult sickle cell patients who received transfusions and who were followed over a 10-year period. STUDY DESIGN AND METHODS: Charts of pediatric and adult sickle cell patients followed at Schneider Children's Hospital (SCH) and Long Island Jewish Medical Center between 1989 and 1999 were retrieved. Patients followed at SCH were classified as pediatric, regardless of age. Data on transfusion history, alloimmunization, and transfusion reactions from 1990 were retrieved from computerized blood bank records. Transfusion history, development of alloantibodies and autoantibodies, and transfusion reactions were correlated with clinical evidence of hemolysis or other adverse reactions from the charts. All patients received ABO- and Rh-compatible blood transfusions for which a partial or extended antigen match was not performed. RESULTS: Among pediatric patients, 29 percent developed clinically significant alloantibodies, and 8 percent developed autoantibodies. Seven patients developed delayed hemolytic and/or serologic transfusion reactions, two with hyperhemolysis, two with clinical evidence of hemolysis, and three with serologic evidence only. The two patients with hyperhemolysis had received extended antigen-matched RBC transfusions to provide blood compatible with their existing antibodies. Among adult patients, 47.0 percent developed significant alloantibodies, and 9.7 percent developed autoantibodies. Five incidences of delayed hemolytic and/or serologic transfusion reactions occurred, one with hyperhemolysis and four with serologic evidence only. CONCLUSION: The alloimmunization rate is 29 percent in pediatric and 47 percent in adult sickle cell patients when partial or extended RBC antigen match is not performed. However, the delayed serologic and/or hemolytic transfusion reactions did not result in severe clinical outcome in most instances. The most important adverse event was hyperhemolysis, which may be triggered by a transfusion, but was not prevented by matching for RBC antigens. In most instances, the cause of hyperhemolysis was multifactorial. PMID- 11896312 TI - Three-base deletion and one-base insertion of the alpha(1,4)galactosyltransferase gene responsible for the P phenotype. AB - BACKGROUND: Recently, an alpha(1,4)galactosyltransferase gene that is responsible for synthesis of P(k) (Gb3) was isolated. The P individuals who did not express the P(k), P, and P(1) antigens on RBC membranes were shown to lack the P(k) (Gb3) synthase activity because of multiple distinct mutations in the alpha(1,4)galactosyltransferase gene. STUDY DESIGN AND METHODS: DNA sequences of the P(k) (Gb3) synthase gene in three Japanese individuals with the p phenotype were analyzed. RESULTS: One individual was found to be homozygous for an allele containing a three-base deletion of CTTCTTC to CTTC from bases 237 through 243 in the coding region. The other two individuals were found to be homozygous for an allele containing a single cytosine insertion in a cytosine repeat at positions 1026 through 1029, resulting in a reading frame shift. CONCLUSION: The P blood group phenotype is due to several distinct nonfunctional alleles without any predominant allele. PMID- 11896314 TI - Higher affinity human D MoAb prepared by light-chain shuffling and selected by phage display. AB - BACKGROUND: In blood banks, D MoAbs are routinely used to phenotype donors and patients. However, most D MoAbs do not agglutinate RBCs that weakly express D. The use of higher affinity MoAbs could overcome this problem. In this work, an attempt has been made to increase the affinity of the human clone 43F10, an IgG anti-D, by light (L)-chain shuffling followed by selection using phage display. STUDY DESIGN AND METHODS: PBMNCs of three polyimmunized individuals were used to construct the kappa L-chain repertoire that was recombined with the 43F10 heavy chain in a phagemid vector system (pComb3H, Scripps Institute, La Jolla, CA). L chain-shuffled 43F10-F(ab) phages were selected on intact RBCs and characterized by ELISA, indirect agglutination, and sequence analysis. RESULTS: L-chain shuffling combined with phage display permitted the selection of a 43F10 MoAb variant (p3.17) with improved reactivity with weak D RBCs in agglutination assays. Nucleic acid sequence analysis showed that p3.17 and wild-type (wt) 43F10 L chains are encoded by different VL segments of the Vk1 family and different J segments, thus showing a relatively low degree of homology (86.4%). CONCLUSION: The use of a variant such as p3.17 could permit a further increase of the potency of existing anti-D reagents. The low homology between p3.17 and wt 43F10 sequences further exemplifies the predominant role of the heavy chain in determining the specificity of the anti-D. PMID- 11896313 TI - Insights into the Holley- and Joseph- phenotypes. AB - BACKGROUND: The Dombrock blood group system consists of two antithetical antigens (Do(a) and Do(b)) and three high-incidence antigens (Gregory [Gy(a)], Holley [Hy], and Joseph [Jo(a)]). Hy and Jo(a) have an unusual phenotypic relationship. All Hy- RBCs are Jo(a-), but not all Jo(a-) RBCs are Hy-. The molecular background associated with Hy- and Jo(a-) phenotypes is reported. STUDY DESIGN AND METHODS: DNA from 18 probands with Gy(a+(w)) Hy- Jo(a-) RBCs (Hy- phenotype) and from 13 probands with Gy(a+) Hy+(w) Jo(a-) RBCs (Jo[a-] phenotype) was tested. RESULTS: Sequencing and PCR-RFLP revealed 323 G>T (Gly 108Val) and 378 T>C (silent mutation) changes on a DOB background (HY) associated with the Hy- samples. The sister of the original Hy- proband and the majority of samples had an additional mutation of 898 C>G (Leu300Val) (HY1); others had 898C (300Leu) (HY2). In the Jo(a-) phenotype, there is a 350 C>T (Thr1 17Ile) and a 378 C>T (silent mutation) change on a DOA background (JO). CONCLUSION: The results provide an explanation for the variation in typing results in antibody producers. The ablation of Jo(a) in the Hy- phenotype and the weakening of Hy in the Jo(a-) phenotype may be due to the close proximity of these antigens. The 898 C>G mutation, within the sequence motif for glycosylphosphatidylinositol linkage, may cause reduced efficiency of anchoring the protein to the RBC membrane, thereby weakening the expression of Gy(a) and Do(b). PMID- 11896315 TI - Detection and removal of fat particles from postoperative salvaged blood in orthopedic surgery. AB - BACKGROUND: Although transfusion or return of salvaged shed blood has become popular in major orthopedic procedures, this blood-saving method is still controversial because shed blood may be contaminated with chemical and tissular debris, such as fat particles, which may increase the risk of fat embolism after bone surgery. STUDY DESIGN AND METHODS: In an effort to find an easy, reliable method for determination of both fat particle content and removal from shed blood, analyses of perioperative blood samples were performed with a cell counter (Technicon H3 [H3]) in orthopedic patients undergoing spinal fusion in which postoperative shed blood was collected and returned with a blood collection canister. A screen or surface filter was intercalated in the return line to eliminate microaggregates, fat particles, and/or WBCs. RESULTS: Fat particles in shed blood are clearly detected as a condensed, sigmoidal-shaped area at the right-hand side of the PMN zone in the channel in which the H3 measures particles according to their degree of lobularity. This signal can be reproduced by the addition of animal or vegetable fat to venous blood, but not by the addition of activated platelets or RBC membranes. Fat particles, together with WBCs and microaggregates, in shed blood were effectively removed by surface filters, whereas screen filters were not effective. CONCLUSION: The use of the TH3 seems to be an easy, reliable, and low-cost approach for monitoring fat particle content and removal from postoperative salvaged shed blood in orthopedic procedures. PMID- 11896316 TI - Storage of platelets in additive solutions: effects of magnesium and/or potassium. AB - BACKGROUND: Previous studies indicate that platelet concentrates (PCs) in a platelet additive solution (PAS) containing citrate, acetate, and sodium chloride (PAS-2) show a significantly higher increase of CD62+ platelets than PCs in other brands of PAS containing Mg(2+) and K(+). To investigate whether this difference can be explained by the presence of Mg(2+) and/or K(+) in the storage medium, we performed paired studies comparing storage of PCs in PAS-2 to PAS-2 with either Mg(2+) or K(+) or both in combination. STUDY DESIGN AND METHODS: PCs from pooled buffy coats were prepared in either PAS-2 or PAS-2 with Mg(2+) or K(+) or both in combination (PAS-2 modified). Different volumes of MgCl(2) solution (1 mol/L) and/or KCl solution (1 mol/L) were added to PAS-2 to obtain various concentrations. After preparation and during storage (at Days 3 and 7), pH, pCO(2), pO(2), HCO(3)(-), and CD62 (%) were measured. RESULTS: During 7 days of storage, pH was very stable (6.9-7.2) in all PCs. At Day 7, platelet CD62 expression was 49 percent (PAS-2), 41 percent (PAS-2 with 1.5 mmol/L Mg(2+)), and 38 percent (PAS-2 with 4.5 mmol/L Mg(2+)). With added K(+), at Day 7, expression of CD62 was 55 percent (PAS-2), 39 percent (PAS-2 with 4.5 mmol/L K(+)), and 35 percent (PAS-2 with 9.0 mmol/L K(+)). In PAS-2 modified (PAS-2 with 1.5 mmol/L Mg(2+) and 4.5 mmol/L K(+)) and CPD plasma, the corresponding CD62 values were 23 and 35 percent, respectively. CONCLUSION: The combination of Mg(2+) and K(+) gave significantly (p < 0.05) lower platelet CD62 expression in the storage medium than in PAS-2. The effects of these differences on platelet metabolism and in vivo properties remain to be investigated. PMID- 11896317 TI - Preparation of FFP as a by-product of plateletpheresis. AB - BACKGROUND: To reduce the production costs of single-donor platelets (SDPs), a study was conducted to investigate whether plasma collected as a by-product of plateletpheresis satisfies the quality requirements for FFP without impairing the quality of the SDP component. STUDY DESIGN AND METHODS: Ninety-two donors with platelet (PLT) counts <270 x 10(9) per L underwent plateletpheresis using an automated cell separator (Spectra Apheresis System with the Leukoreduction System [LRS], Gambro BCT, Lakewood, CO). The machine was programmed to collect 3 x 10(11) PLTs in 250 mL of plasma with an additional unit of 350 mL of plasma or 3 x 10(11) PLTs in 250 mL of plasma without additional plasma in 10 procedures. FV and FVIII and residual RBCs, WBCs, and PLTs in the plasma were measured for quality control. RESULTS: FV was 0.87 +/- 0.18 IU per mL, and FVIII was 1.32 +/- 0.48 IU per mL in the plasma components (n = 41). The recovery was 94.1 +/- 5.5 percent for FV and 102.2 +/- 9.5 percent for FVIII when compared with the donors' predonation values. Residual cells were 0.002 +/- 0.009 x 10(9) RBCs per L (n = 30), 12 +/- 6 x 10(9) PLTs per L (n = 30), and 0.32 +/- 0.37 x 10(6) WBCs per L (n = 92). CONCLUSIONS: Using the automated cell separator and special software, it is possible to collect plasma as a by-product of plateletpheresis that meets the properties requested for FFP without impairing the quality of the SDP components. The content of clotting factors is within the requested range for FFP. Residual cell counts are within all European and U.S. specifications for FFP, and the WBC content even satisfies the criteria for WBC-reduced blood components. The collection of FFP as a by-product does not cause any additional costs and thus helps to reduce the costs in preparing blood components. PMID- 11896318 TI - Quantitation of residual WBCs in filtered blood components by high-throughput, real-time kinetic PCR. AB - BACKGROUND: The effort to eliminate transfusion complications associated with WBCs has led to the widespread use of filters able to reduce WBC concentrations to 54 (OR, 1.6; 95% CI, 1.3-2.1), age >66 (OR, 2.7; 95% CI, 2.0-3.5), female sex (OR, 1.6; 95% CI, 1.2-2.0), diabetes with chronic complications (OR, 2.5; 95% CI, 1.3-4.9), and metastatic tumor (OR, 4.9; 95% CI, 2.3-10.5), emergency room admission (OR, 2.3; 95% CI, 1.4-3.8), and greater hospital volume (OR, 4.0; 95% CI, 1.8-8.6). Characteristics independently associated with increased autologous transfusions (n = 574) included white race (OR, 1.7; 95% CI, 1.2-2.4), female sex (OR, 1.4; 95% CI, 1.1-1.8), and greater surgeon volume (OR, 3.5; 95% CI, 1.4 9.1). DISCUSSION: This information can be used to provide informed risk-benefit discussions with patients regarding the risk for blood transfusion as well as to target high-risk patients and institutions for interventions to reduce the risk of exposure to blood components. PMID- 11896334 TI - Pharmacokinetic analysis of plasma-derived and recombinant F IX concentrates in previously treated patients with moderate or severe hemophilia B. AB - BACKGROUND: Hemophilia B is an X-linked bleeding disorder that affects approximately 1 in 25,000 males. Therapy for acute bleeding episodes consists of transfusions of plasma-derived (pd-F IX) or recombinant (r-F IX) concentrates. STUDY DESIGN AND METHODS: A double-blind, two-period crossover study was initiated to assess the pharmacokinetics of pd-F IX and r-F IX and to address patient-specific variables that might influence in vivo recovery. Study product was administered by a single bolus infusion (50 IU/kg) to 43 previously treated patients in the nonbleeding state, and F IX:C levels were measured over a period of 48 hours after infusion. RESULTS: The mean in vivo recovery in the pd-F IX group was 1.71 +/- 0.73 IU per dL per IU per kg compared with 0.86 +/- 0.31 IU per dL per IU per kg with r-F IX (p 0.88. For both the Harrison modified Risser-Ferguson method on AP views and posterior tangent method on lateral cervical views, the mean absolute differences of observers' measurements are small. PMID- 11896377 TI - Long-term effectiveness of bone-setting, light exercise therapy, and physiotherapy for prolonged back pain: a randomized controlled trial. AB - BACKGROUND: Chiropractic manipulation and strenuous exercise therapy have been shown effective in the treatment of nonspecific back pain. Bone-setting, the predecessor of modern manual therapies, still survives in some parts of Finland and was compared with a light exercise therapy and non-manipulative, pragmatic physiotherapy in a year-long randomized controlled trial on patients with long term back pain. METHODS: One hundred fourteen ambulatory patients of working age with back pain for 7 weeks or more were randomly assigned to the therapies, which were offered in up to 10 sessions during a 6-week treatment period. The outcome was measured by the Oswestry Disability Questionnaire. Sick-leaves and visits to health centers were recorded for 1 year before and after the therapy. RESULTS: The Oswestry disability scores improved most in the bone-setting group (P =.02, Kruskall-Wallis test). Visits to health centers for back pain were reduced only in the physiotherapy group (P =.01, Wilcoxon test). Sick-leaves were not significantly different between groups. A secondary analysis based on the use of additional therapies after the intervention showed a possible subgroup with an enhanced effect from bone-setting. CONCLUSIONS: Traditional bone-setting seemed more effective than exercise or physiotherapy on back pain and disability, even 1 year after therapy. PMID- 11896378 TI - The minimum energy hypothesis: a unified model of fixation resolution. AB - OBJECTIVE: To present a new theoretical construct, the Minimum Energy Hypothesis, which explains structural changes observed in the spine concomitant to spinal joint fixation resolution in initial investigations. DESIGN: Theoretical analysis. HYPOTHESIS: A unified theory of manipulative effectiveness is proposed that integrates the fixation and sensory tonus models of manipulation. The theory is based on the fact that the spine will assume a position of minimum internal energy when mechanical equilibrium is achieved. By using a simple mathematical model, it is shown that the fixation model and the sensory tonus models are 2 different aspects of the same theoretical construct. The Minimum Energy Hypothesis predicts that the spine will seek an optimal minimum energy configuration if the constraints preventing it from doing so are removed. Constraints are hypothesized to be joint fixations caused by inflammation in and about the spine and its sequella, muscle spasm, fibroadipose and scar tissue, and ultimately, degeneration. It is further hypothesized that the use of a computerized mechanical manipulative device may resolve such fixations, an example of which is radiographically demonstrable cervical hypolordosis. CONCLUSION: A unified theory of manipulative effectiveness based on the concept of minimum energy to attain mechanical equilibrium is brought forward to explain the results of initial investigations. PMID- 11896379 TI - Incidence of foot rotation, pelvic crest unleveling, and supine leg length alignment asymmetry and their relationship to self-reported back pain. AB - OBJECTIVE: To determine the incidence of pelvic unleveling, foot rotation, and supine leg length alignment asymmetry in a nonclinical population and to examine the validity (sensitivity, specificity, positive and negative predictive values) of these visual tests and their relationship to self-reported back pain. DESIGN: Volunteers answered a questionnaire regarding back pain and were then examined by a chiropractor who was unaware of the status of their back pain. PARTICIPANTS: Seventy-four unscreened volunteers answered the questionnaire. MAIN OUTCOME MEASURES: The association of visual tests with back pain and their validity indices; Visual Analogue Scale ratings. RESULTS: Fifty-one percent (n = 74) of volunteers examined had supine leg length alignment asymmetry (LLA). Pain intensity on a Visual Analogue Scale was significantly higher (P <.001) for those demonstrating supine LLA than for those without LLA. Those with back pain and recurrent back pain were significantly (P <.001) more likely to have supine LLA. The validity indices of the supine leg check showed acceptable levels for sensitivity (74%), specificity (78%), and positive predictive value (82%) [corrected] in recurrent back pain. Findings also indicated a high incidence of supine LLA in volunteers with chronic back pain (85%). CONCLUSION: The results indicated that, in this group of volunteers, the supine leg length alignment check had clinical validity as a stand-alone test for recurring back pain. Further testing on a larger, statistically defined cross-section of the population is recommended. PMID- 11896380 TI - Certification examinations for massage therapists: a psychometric analysis. AB - OBJECTIVE: To describe the components of the Alberta Registered Massage Therapists Society (ARMTS) examination and their psychometric properties. METHODS: All 3 components of the ARMTS examination (knowledge, clinical judgement, and clinical skills) were administered to 112 candidates. The examination consisted of 2 written components (140 multiple-choice questions on basic science knowledge and 60 multiple-choice questions on clinical judgment) and a clinical competency assessment of the following practical skills with standardized patients: (1) taking a case history, (2) assessing physical condition, and (3) treating the condition. All components of the examination were criterion-referenced with the methods of minimum performance level (MPL). RESULTS: The internal consistency reliability coefficients (Cronbach alpha) ranged from 0.60 to 0.88 for all test components. The descriptive statistics, performance levels, and reliability estimates indicate that the examination is functioning well. Concurrent, criterion-related validity evidence was provided by correlations between the examination components that ranged from r = 0.24 (P <.05) to r = 0.78 (P <.01). Factor analysis produced 3 factors: information processing, clinical treatment, and follow-up management. CONCLUSIONS: The results provide evidence of adequate-to-good internal consistency reliability and content validity. Empirical validity evidence based on concurrent, criterion related measures is provided by the correlational analysis. The significant correlations indicate that although performance is related across the examination, the various components do assess unique and independent domains. This is further supported by the results of the factor analysis that provide evidence for discriminant validity of the measures (ie, they discriminate between domains of measurement such as information processing, treatment, and basic knowledge). Taken together, these results indicate that the ARMTS examination has evidence for both reliability and validity. PMID- 11896381 TI - Evolution of foot orthotics--part 1: coherent theory or coherent practice? AB - OBJECTIVE: To present a critical review of the evolution of foot orthotics theory and clinical practice. DATA SOURCES: Several classic publications were consulted because of their overwhelming influence. The work of Merton L. Root and his colleagues in the 1970s was carefully examined. Careful evaluations were performed to determine how faithfully Root's central concepts were subsequently followed. Studies attempting to validate this and other orthotic paradigms were also reviewed. RESULTS: Epidemiologic studies provide strong support for the clinical advantages of orthoses, yet explanations of foot orthotic mechanisms remain elusive. Considerable variability has crept into the literature with respect to Root's core theoretical concepts of how and why to determine the neutral position of the subtalar joint (weight-bearing vs non-weight-bearing, palpation vs range-checking). Numerous studies document poor clinical reliability and validity; indeed, this paradigm appears to favor supination, thereby violating its "neutral" premise. Mechanisms other than those of the classic Root theory must be at work. Accordingly, successes have been achieved with alternate paradigms that use much simpler casting techniques. Although less frequently cited, successes have been gained with various viscoelastic materials that enhance shock-absorption and proprioception, as well as custom-made flexible orthotic designs that emphasize the 3 natural arches of the foot. CONCLUSIONS: The use of foot orthoses is well documented for the treatment of many maladies, yet clinical successes have been achieved both inside and outside of the classic Root paradigm. Clearly, a more complete theoretical understanding of the mechanisms of foot orthotics awaits discovery. PMID- 11896382 TI - Evolution of foot orthotics--part 2: research reshapes long-standing theory. AB - OBJECTIVE: To challenge casual understanding of the causal mechanisms of foot orthotics. Although the classic orthotic paradigm of Merton L. Root and his colleagues is often acknowledged, the research attempting to explain and validate these mechanisms is far less clear in its appraisal. DATA SOURCES: Studies evaluating the relationship of foot type (medial arch height) and use of foot orthoses to the motions of the foot and ankle were compared and contrasted. A search was conducted to evaluate other possible mechanisms of orthotic intervention. RESULTS: Although Root's methods of foot evaluation (subtalar neutral position) and casting (non-weight-bearing) are well referenced, these methods have poor reliability, unproven validity, and are, in fact, seldom strictly followed. We challenge 2 widely held concepts: that excessive foot eversion leads to excessive pronation and that orthotics provide beneficial effects by controlling rearfoot inversion/eversion. Numerous studies show that patterns of rearfoot inversion/eversion cannot be characterized either by foot type or by orthotics use. Rather, subtle control of internal/external tibial rotation appears to be the most significant factor in maintaining proper supination/pronation mechanics. Recent evidence also suggests that proprioceptive influences play a large, and perhaps largely unexplored, role. CONCLUSIONS: Considerable evidence supports the exploration of new theories and paradigms of orthotics use. Investigations of flexible orthotic designs, proprioceptive influences, and the 3-dimensional effects of subtalar joint motion on the entire kinetic chain are areas of research that show great promise. PMID- 11896383 TI - Chronic pain/dysfunction in whiplash-associated disorders. PMID- 11896385 TI - Medically supervised water-only fasting in the treatment of hypertension. PMID- 11896387 TI - Measuring the sacral inclination angle in clinical practice: is there an alternative to radiographs? PMID- 11896389 TI - Haploinsufficiency of NSD1 causes Sotos syndrome. AB - We isolated NSD1 from the 5q35 breakpoint in an individual with Sotos syndrome harboring a chromosomal translocation. We identified 1 nonsense, 3 frameshift and 20 submicroscopic deletion mutations of NSD1 among 42 individuals with sporadic cases of Sotos syndrome. The results indicate that haploinsufficiency of NSD1 is the major cause of Sotos syndrome. PMID- 11896390 TI - Micro RNAs are complementary to 3' UTR sequence motifs that mediate negative post transcriptional regulation. AB - Micro RNAs are a large family of noncoding RNAs of 21-22 nucleotides whose functions are generally unknown. Here a large subset of Drosophila micro RNAs is shown to be perfectly complementary to several classes of sequence motif previously demonstrated to mediate negative post-transcriptional regulation. These findings suggest a more general role for micro RNAs in gene regulation through the formation of RNA duplexes. PMID- 11896391 TI - How antibodies to a ubiquitous cytoplasmic enzyme may provoke joint-specific autoimmune disease. AB - Arthritis in the K/BxN mouse model results from pathogenic immunoglobulins (Igs) that recognize the ubiquitous cytoplasmic enzyme glucose-6-phosphate isomerase (GPI). But how is a joint-specific disease of autoimmune and inflammatory nature induced by systemic self-reactivity? No unusual amounts or sequence, splice or modification variants of GPI expression were found in joints. Instead, immunohistological examination revealed the accumulation of extracellular GPI on the lining of the normal articular cavity, most visibly along the cartilage surface. In arthritic mice, these GPI deposits were amplified and localized with IgG and C3 complement. Similar deposits were found in human arthritic joints. We propose that GPI-anti-GPI complexes on articular surfaces initiate an inflammatory cascade via the alternative complement pathway, which is unbridled because the cartilage surface lacks the usual cellular inhibitors. This may constitute a generic scenario of arthritogenesis, in which extra-articular proteins coat the cartilage or joint extracellular matrix. PMID- 11896392 TI - TLR4, but not TLR2, mediates IFN-beta-induced STAT1alpha/beta-dependent gene expression in macrophages. AB - Toll-like receptor 2 (TLR2) agonists induce a subset of TLR4-inducible proinflammatory genes, which suggests the use of differential signaling pathways. Murine macrophages stimulated with the TLR4 agonist Escherichia coli lipopolysaccharide (LPS), but not with TLR2 agonists, induced phosphorylation of signal transducer and activator of transcription 1alpha (STAT1alpha) and STAT1beta, which was blocked by antibodies to interferon beta (IFN-beta) but not IFN-alpha. All TLR2 agonists poorly induced IFN-beta, which is encoded by an immediate early LPS-inducible gene. Thus, the failure of TLR2 agonists to induce STAT1-dependent genes resulted, in part, from their inability to express IFN beta. TLR4-induced IFN-beta mRNA was MyD88- and PKR (double-stranded RNA dependent protein kinase)-independent, but TIRAP (Toll-interleukin 1 receptor domain-containing adapter protein)-dependent. Together, these findings provide the first mechanistic basis for differential patterns of gene expression activated by TLR4 and TLR2 agonists. PMID- 11896393 TI - Dynamic visualization of a joint-specific autoimmune response through positron emission tomography. AB - In the K/BxN mouse model of rheumatoid arthritis, the transfer of autoantibodies specific for glucose-6-phosphate isomerase (GPI) into naive mice rapidly induces joint-specific inflammation similar to that seen in human rheumatoid arthritis. The ubiquitous expression of GPI and the systemic circulation of anti-GPI immunoglobulin G (IgG) seem incongruous with the tissue specificity of this disease. By using PET (positron emission tomography), we show here that purified anti-GPI IgG localizes specifically to distal joints in the front and rear limbs within minutes of intravenous injection, reaches saturation by 20 min and remains localized for at least 24 h. In contrast, control IgG does not localize to joints or cause inflammation. The rapid kinetics of anti-GPI IgG joint localization supports a model in which autoantibodies bind directly to pre-existing extracellular GPI in normal healthy mouse joints. PMID- 11896394 TI - Role of antigen receptor affinity in T cell-independent antibody responses in vivo. AB - To examine how B cell receptor affinity affects clonal selection in thymus independent type 2 (TI-2) immune responses, we produced mice with antibodies that showed a 40-fold difference in affinity for the hapten (4-hydroxy-3 nitrophenyl)acetyl (NP). The difference in the responses of high- and low affinity B cells to NP-Ficoll was only twofold. However, in competition experiments only the high-affinity B cells responded to antigen. CD19 deficiency increased the affinity threshold of TI-2 responses, whereas Lyn deficiency enhanced clonal expansion but abrogated B cell terminal differentiation. Thus, in TI-2 immune responses, large differences in affinity produce only small differences in the intrinsic ability of B cells to respond to antigen, and selection for high-affinity clones is due to clonal competition during the earliest stages of the response. PMID- 11896395 TI - Cooperation between independent hippocampal synapses is controlled by glutamate uptake. AB - Localized action of released neurotransmitters is the basis for synaptic independence. In the hippocampal neuropil, where synapses are densely packed, it has been postulated that released glutamate, by diffusing out of the synaptic cleft, may also activate postsynaptic receptors at neighboring synapses. Here we show that neighboring excitatory synapses on hippocampal CA1 pyramidal cells can cooperate in the activation of postsynaptic receptors through the confluence of released glutamate, and that this cooperation is controlled by glutamate uptake. Furthermore, glutamate transporters control temporal interactions between transmitter transients originating from the same axon. Thus, cooperative interactions between excitatory synapses are modulated in space and time by glutamate uptake. PMID- 11896396 TI - The information efficacy of a synapse. AB - We provide a functional measure, the synaptic information efficacy (SIE), to assess the impact of synaptic input on spike output. SIE is the mutual information shared by the presynaptic input and postsynaptic output spike trains. To estimate SIE we used a method based on compression algorithms. This method detects the effect of a single synaptic input on the postsynaptic spike output in the presence of massive background synaptic activity in neuron models of progressively increasing realism. SIE increased with increases either in time locking between the input synapse activity and the output spike or in the average number of output spikes. SIE depended on the context in which the synapse operates. We also measured SIE experimentally. Systematic exploration of the effect of synaptic and dendritic parameters on the SIE offers a fresh look at the synapse as a communication device and a quantitative measure of how much the dendritic synapse informs the axon. PMID- 11896397 TI - Voluntary action and conscious awareness. AB - Humans have the conscious experience of 'free will': we feel we can generate our actions, and thus affect our environment. Here we used the perceived time of intentional actions and of their sensory consequences as a means to study consciousness of action. These perceived times were attracted together in conscious awareness, so that subjects perceived voluntary movements as occurring later and their sensory consequences as occurring earlier than they actually did. Comparable involuntary movements caused by magnetic brain stimulation reversed this attraction effect. We conclude that the CNS applies a specific neural mechanism to produce intentional binding of actions and their effects in conscious awareness. PMID- 11896398 TI - Glial cells generate neurons: the role of the transcription factor Pax6. AB - Radial glial cells, ubiquitous throughout the developing CNS, guide radially migrating neurons and are the precursors of astrocytes. Recent evidence indicates that radial glial cells also generate neurons in the developing cerebral cortex. Here we investigated the role of the transcription factor Pax6 expressed in cortical radial glia. We showed that radial glial cells isolated from the cortex of Pax6 mutant mice have a reduced neurogenic potential, whereas the neurogenic potential of non-radial glial precursors is not affected. Consistent with defects in only one neurogenic lineage, the number of neurons in the Pax6 mutant cortex in vivo is reduced by half. Conversely, retrovirally mediated Pax6 expression instructs neurogenesis even in astrocytes from postnatal cortex in vitro. These results demonstrated an important role of Pax6 as intrinsic fate determinant of the neurogenic potential of glial cells. PMID- 11896399 TI - Dendritic spines do not split during hippocampal LTP or maturation. PMID- 11896400 TI - Efficient coding of natural sounds. AB - The auditory system encodes sound by decomposing the amplitude signal arriving at the ear into multiple frequency bands whose center frequencies and bandwidths are approximately exponential functions of the distance from the stapes. This organization is thought to result from the adaptation of cochlear mechanisms to the animal's auditory environment. Here we report that several basic auditory nerve fiber tuning properties can be accounted for by adapting a population of filter shapes to encode natural sounds efficiently. The form of the code depends on sound class, resembling a Fourier transformation when optimized for animal vocalizations and a wavelet transformation when optimized for non-biological environmental sounds. Only for the combined set does the optimal code follow scaling characteristics of physiological data. These results suggest that auditory nerve fibers encode a broad set of natural sounds in a manner consistent with information theoretic principles. PMID- 11896401 TI - Inhibition of p38 MAP kinase by utilizing a novel allosteric binding site. AB - The p38 MAP kinase plays a crucial role in regulating the production of proinflammatory cytokines, such as tumor necrosis factor and interleukin-1. Blocking this kinase may offer an effective therapy for treating many inflammatory diseases. Here we report a new allosteric binding site for a diaryl urea class of highly potent and selective inhibitors against human p38 MAP kinase. The formation of this binding site requires a large conformational change not observed previously for any of the protein Ser/Thr kinases. This change is in the highly conserved Asp-Phe-Gly motif within the active site of the kinase. Solution studies demonstrate that this class of compounds has slow binding kinetics, consistent with the requirement for conformational change. Improving interactions in this allosteric pocket, as well as establishing binding interactions in the ATP pocket, enhanced the affinity of the inhibitors by 12,000 fold. One of the most potent compounds in this series, BIRB 796, has picomolar affinity for the kinase and low nanomolar inhibitory activity in cell culture. PMID- 11896402 TI - Two-metal active site binding of a Tn5 transposase synaptic complex. AB - A synaptic complex of Tn5 transposase with an extended outside end DNA duplex was prepared and crystallized, and its crystal structure was determined in an effort to reveal the role of metal ions in catalysis. Two Mn2+ ions bound to the active site when a single nucleotide of donor DNA was added to the 3' end of the transferred strand. Marked conformational changes were observed in the DNA bases closest to the active site. The position of the metal ions and the conformational changes of the DNA provide insight into the mechanism of hairpin formation and cleavage, and is consistent with a two-metal model for catalysis. PMID- 11896403 TI - Cysteine-rich module structure reveals a fulcrum for integrin rearrangement upon activation. AB - Cysteine-rich repeats in the integrin beta subunit stalk region relay activation signals to the ligand-binding headpiece. The NMR solution structure and disulfide bond connectivity of Cys-rich module-3 of the integrin beta2 subunit reveal a nosecone-shaped variant of the EGF fold, termed an integrin-EGF (I-EGF) domain. Interdomain contacts between I-EGF domains 2 and 3 observed by NMR support a model in which the modules are related by an approximate two-fold screw axis in an extended arrangement. Our findings complement a 3.1 A crystal structure of the extracellular portion of integrin alphaVbeta3, which lacks an atomic model for I EGF2 and a portion of I-EGF3. The disulfide connectivity of I-EGF3 chemically assigned here differs from the pairings suggested in the alphaVbeta3 structure. Epitopes that become exposed upon integrin activation and residues that restrain activation are defined in beta2 I-EGF domains 2 and 3. Superposition on the alphaVbeta3 structure reveals that they are buried. This observation suggests that the highly bent alphaVbeta3 structure represents the inactive conformation and that release of contacts with I-EGF modules 2 and 3 triggers a switchblade like opening motion extending the integrin into its active conformation. PMID- 11896404 TI - Crystal structure of a transition state mimic of the catalytic subunit of cAMP dependent protein kinase. AB - To understand the molecular mechanism underlying phosphoryl transfer of cAMP dependent protein kinase, the structure of the catalytic subunit in complex with ADP, aluminum fluoride, Mg2+ ions and a substrate peptide was determined at 2.0 A resolution. Aluminum fluoride was modeled as AlF3 in a planar geometry; it is positioned 2.3 A from both the donor oxygen of ADP and the hydroxyl group of the recipient Ser residue. In this configuration, the aluminum atom forms a trigonal bipyramidal coordination with the oxygen atoms of the donor and recipient groups at the apical positions. This arrangement suggests that aluminum fluoride mimics the transition state and provides the first direct structural evidence for the in line mechanism of phosphoryl transfer in a protein kinase. PMID- 11896405 TI - Prevalence of putative periodontopathogens from periodontal patients and healthy subjects in Sao Paulo, SP, Brazil. AB - A. actinomycetemcomitans, B. forsythus, P. gingivalis, C. rectus, E. corrodens, P. intermedia, F. nucleatum, and T. denticola were identified from subgingival plaque from 50 periodontal patients and 50 healthy subjects. PCR products from each species showed a specific band and could be used to identify periodontal organisms from clinical specimens. Identical negative or positive results between PCR and culture occurred in 66% (A. actinomycetemcomitans) to 93% (F. nucleatum) of the samples. PCR detection odds ratio values for A. actinomycetemcomitans, B. forsythus, C. rectus, E. corrodens, P. intermedia, and T. denticola were significantly associated with disease having a higher OR values for B. forsythus (2.97, 95% CI 1.88 - 4.70). Cultures showed that A. actinomycetemcomitans, B. forsythus and P. intermedia were associated with periodontitis, however, P. gingivalis, C. rectus, E. corrodens and F. nucleatum were not significantly associated with the disease. PMID- 11896406 TI - Frequency of serum anti-cysticercus antibodies in the population of a rural Brazilian community (Cassia dos coqueiros, SP) determined by Elisa and immunoblotting using Taenia crassiceps antigens. AB - Considering the impact of cysticercosis on public health, especially the neurologic form of the disease, neurocysticercosis (NC), we studied the frequency of positivity of anti-Taenia solium cysticercus antibodies in serum samples from 1,863 inhabitants of Cassia dos Coqueiros, SP, a municipal district located 80 km from Ribeirao Preto, an area considered endemic for cysticercosis. The 1,863 samples were tested by enzyme linked immunosorbent assay (ELISA) using an antigenic extract from Taenia crassiceps vesicular fluid (Tcra). The reactive and inconclusive ELISA samples were tested by immunoblotting. Of the 459 samples submitted to immunoblotting, 40 were strongly immunoreactive to the immunodominant 18 and 14 kD peptides. Considering the use of immunoblotting as confirmatory due to its high specificity, the anti-cysticercus serum prevalence in this population was 2.1%. PMID- 11896408 TI - Association between human parvovirus B19 and arthropathy in Belem, Para, North Brazil. AB - A total of 220 patients with arthropathy were selected in Belem, Para between January 1994 and December 2000, and screened for the presence of human parvovirus B19 IgM and IgG antibodies by enzyme-linked immunosorbent assay (ELISA). A subgroup (n = 132) of patients with high levels of antibodies (either IgM+/IgG+ or IgM-/IgG+) were examined for the presence of DNA by polymerase chain reaction/nested PCR. Recent/active infection (detection of IgM and/or IgG specific antibodies and presence of viral DNA) was identified in 47.7% of the 132 individuals with arthropathy. In our study, women were significantly more affected (59.7%) than men (35.4%) (P = 0.0006). The age group of 11-20 years (84.6%), among female patients, and 21-30 years (42.1%), among male, were those with the highest incidence rates. The analysis of the temporal distribution of B19-associated arthropaties showed a cyclic pattern, with peak incidence rates occuring at 3-5 year intervals. Significant diference (P = 0.01) was observed when comparing both the highest (39.0%) and the lowest (11.0%) seropositivity rates for the years of 1995 and 2000, respectively. The interfalangial joints of hands and feet were mostly affected, with 50.0% and 48.0% of cases among both women and men, respectively. In a smaller proportion, other joints such as those of knee, ankle, pulse and shoulder were affected. As for the duration, symptoms lasted 1 to 5 days in 54.0% of the individuals, whereas in 46.0% of them the disease lasted 6-10 days, if considered the subgroup (n = 63) of patients with recent/active infection by parvovirus B19. In our study, joint clinical manifestations occurred symmetrically. Our results indicate that B19 may be an important agent of arthropathies in our region, and this underscores the need for specific laboratory diagnosis when treating patients suffering from acute arthropathy, mainly pregnant women. PMID- 11896407 TI - Nutritional status in relation to the efficacy of the rhesus-human reassortant, tetravalent rotavirus vaccine (RRV-TV) in infants from Belem, para state, Brazil. AB - The rhesus-human reassortant, tetravalent rotavirus vaccine (RRV-TV) was licensed for routine use in the United States of America but it was recently withdrawn from the market because of its possible association with intussusception as an adverse event. The protective efficacy of 3 doses of RRV-TV, in its lower-titer (4 x 10(4) pfu/dose) formulation, was evaluated according to the nutritional status of infants who participated in a phase III trial in Belem, Northern Brazil. A moderate protection conferred by RRV-TV was related to weight-for-age Z scores (WAZ) greater than -1 only, with rates of 38% (p = 0.04) and 40% (p = 0.04) for all- and- pure rotavirus diarrhoeal cases, respectively. In addition, there was a trend for greater efficacy (43%, p = 0.05) among infants reaching an height-for-age Z-score (HAZ) of > -1. Taking WAZ, HAZ and weight-for-height Z score (WHZ) indices 0.05) if both placebo and vaccine groups are compared. There was no significant difference if rates of mixed and pure rotavirus diarrhoeal cases are compared in relation to HAZ, WAZ and weight-for-height Z-score (WHZ) indices. Although a low number of malnourished infants could be identified in the present study, our data show some evidence that malnutrition may interfere with the efficacy of rotavirus vaccines in developing countries. PMID- 11896409 TI - Phlebotominae fauna in the province of Tucuman, Argentina. AB - American Tegumentary Leishmaniasis is endemic in the province of Tucuman since 1916 where the first Argentinian case of leishmaniasis was reported. An epidemic outbreak took place in the province during 1986-1988, after that the number of cases increased gradually again between 1991-1996. Since October of 1999 sand flies captures were performed at different places associated with current and past transmission or sites identified as risky ones. The collections were carried out with CDC mini light trap and modified Shannon trap. In this study 2338 Phlebotominae were captured being prevalent Lutzomyia neivai (Lu intermedia complex) (97.7%), followed by Lu. migonei. We report Lu. shannoni for the first time in the province. Phlebotominae was abundant in the warm and humid months and showed a peak before the summer rains. Lutzomyia neivai showed a pattern (peridomestic prevalence, anthropophilia, abundance in secondary forest) consistent with other outbreak studies. Thus, the results reinforce this species incrimination as vector of leishmaniasis in the area. Domestic animals close to houses increase its abundance, and so the probable associated risk of human Phlebotominae contact. Further studies should be done to understand the role of each Phlebotominae species in the transmission of leishmaniasis in Tucuman in order to design entomological surveillance strategies. PMID- 11896410 TI - Long term evaluation of etiological treatment of chagas disease with benznidazole. AB - The aim of this article is to present an investigation of cure rate, after long follow up, of specific chemotherapy with benznidazole in patients with both acute and chronic Chagas disease, applying quantitative conventional serological tests as the base of the criterion of cure. Twenty one patients with the acute form and 113 with one or other of the various chronic clinical forms of the disease were evaluated, after a follow up period of 13 to 21 years, for the acute, and 6 to 18 years, for the chronic patients. The duration of the acute as well as the chronic disease, a condition which influences the results of the treatment, was determined. The therapeutic schedule was presented, with emphasis on the correlation between adverse reactions and the total dose of 18 grams, approximately, as well as taking into consideration precautions to assure the safety of the treatment. Quantitative serological reactions consisting of complement fixation, indirect immunofluorescence, indirect hemagglutination, and, occasionally, ELISA, were used. Cure was found in 76 per cent of the acute patients but only in 8 per cent of those with chronic forms of the disease. In the light of such contrasting results, fundamentals of the etiological therapy of Chagas disease were discussed, like the criterion of cure, the pathogenesis and the role of immunosuppression showing tissue parasitism in long standing chronic disease, in support of the concept that post-therapeutic consistently positive serological reactions mean the presence of the parasite in the patient's tissues. In relation to the life-cycle of T. cruzi in vertebrate host, there are still some obscure and controversial points, though there is no proof of the existence of resistant or latent forms. However, the finding over the last 15 years, that immunosuppression brings about the reappearance of acute disease in long stand chronic patients justifies a revision of the matter. Facts were quoted in favor of the treatment of chronic patients. PMID- 11896411 TI - Molecular aspects of hepatic carcinogenesis. AB - Exogenous agents correlated with hepatocellular carcinoma (HCC) have been identified and well characterized. These agents, including the different viruses that cause chronic hepatitis and cirrhosis, can lead to regenerative nodules and dysplastic nodules/adenomatous hyperplasia. These conditions associated with several molecular alterations of hepatocyte ultimately culminate in hepatocellular carcinoma. Recently, there has been a great progress in the identification of somatic and germinative mutations that may be correlated with the development of HCC, justifying a review on the subject. Hence, the factors involved in the process of hepatic carcinogenesis, such as infection by the hepatitis B and C viruses, with a special focus in the molecular alterations described in recent years are discussed herein, pointing out areas potentially relevant for clinical development. PMID- 11896412 TI - Parinaud'S oculoglandular syndrome associated with paracoccidioidomycosis. AB - The authors report one case of Parinaud's oculoglandular syndrome associated with Paracoccidioides brasiliensis infection. No other medical report of this condition was found in the medical literature available at Index Medicus and Medline. The eye involvement has been rather uncommon in paracoccidioidomycosis and this report emphasizes the possibility of this kind of presentation making it also necessary to include paracoccidioidomycosis among the several known causes of Parinaud's oculoglandular syndrome. PMID- 11896413 TI - Natural infection of deroceras laeve (Mollusca: gastropoda) with metastronbylid larvae in a transmission focus of abdominal angiostrongyliasis. AB - Angiostrongylus costaricensis is a nematode parasitic of rodents. Man may become infected by ingestion of the third stage larvae produced within the intermediate hosts, usually slugs from the family Veronicellidae. An epidemiological study carried out in a locality in southern Brazil (western Santa Catarina State) where these slugs are a crop pest and an important vector for A. costaricensis has documented for the first time the natural infection of Deroceras laeve with metastrongylid larvae. This small limacid slug is frequently found amid the folds of vegetable leaves and may be inadvertently ingested. Therefore D. laeve may have an important role in transmission of A. costaricensis to man. PMID- 11896414 TI - Dogs may be a reservoir host for Angiostrongylus costaricensis. AB - Angiostrongylus costaricensis is a parasitic nematode of wild rodents. Several other vertebrate species including man may become infected by ingestion of the third stage larvae produced by the intermediate hosts, usually slugs from the family Veronicellidae. There is a report of the diagnosis of abdominal angiostrongyliasis in Canis familiaris with lesions resembling those found in human disease. As a preliminar evaluation of the adequacy of a canine model for pathogenetic studies, a dog was inoculated with 75 L3 of A. costaricensis. Infection was established and fist stage larvae were found in feces up to 88 days post infection, sometimes in very large numbers (9.5 x 10(4) L1/g). No clinical manifestations or significant lesions were detected. These are indications that dog may play a role as a reservoir host for A. costaricensis. PMID- 11896415 TI - A rapid latex agglutination test for the detection of anti-cysticercus antibodies in cerebrospinal fluid (CSF). AB - Simple and rapid latex-based diagnostic tests have been used for detecting specific antigens or antibodies in several diseases. In this article, we present the preliminary results obtained with a latex agglutination test (LAT) for diagnosing neurocysticercosis by detection of antibodies in CSF. A total of 43 CSF samples were assayed by the LAT: 19 CSF samples from patients with neurocysticercosis and 24 CSF samples from patients with other neurologic disorders (neurosyphilis, n = 8; neurotoxoplasmosis, n = 3; viral meningitis, n = 4, chronic headache, n = 9). The LAT exhibited 89.5% sensitivity and 75% specificity. The use of LAT seems to be an additional approach for the screening of neurocysticercosis with advantage of simplicity and rapidity. Further studies could be performed using purified antigens and serum samples. PMID- 11896416 TI - Effects of finasteride on apoptosis and regulation of the human hair cycle. AB - BACKGROUND: A number of studies have provided evidence that apoptosis is a central element in the regulation of hair follicle regression. In androgenetic alopecia (AGA), the exact location and control of key players in the apoptotic pathways remains obscure. OBJECTIVE: In the present study, we used a panel of antibodies and investigated the spatial and cellular pattern of expression of caspases and inhibitors of apoptosis (IAPs), such as XIAP and FLIP, in men with normal scalp and in men with AGA before and after 6 months of treatment with 1 mg oral finasteride treatment. METHODS AND RESULTS: Constitutive expression of caspases-1, -3, -8, and -9 and XIAP was detected predominantly within the isthmic and infundibular hair follicle area, basilar layer of the epidermis, and eccrine and sebaceous glands. AGA-affected tissues showed an increase in caspase (-1, -3, -6, -9) immunoreactivity with a concomitant decrease in XIAP staining. After 6 months of finasteride treatment, both caspases and XIAP were similar to levels exhibited by normal subjects. Immunoblot analysis was performed to determine antibody specificity and cellular expression of caspases. Purified populations of keratinocytes, melanocytes, dermal papilla, and dermal fibroblasts derived from human hair follicles were cultured in vitro and treated with 0.5 mm staurosporin. Time-course experiments revealed that processing of caspase-3 is a principal event during apoptosis of these hair cell types. CONCLUSION: These data suggest that alterations in levels of caspases and IAPs regulate hair follicle homeostasis. Moreover, finasteride appears to influence caspase and XIAP expression in hair follicle cells thus signaling anagen, active growth in the hair cycle. PMID- 11896417 TI - Skin-related quality of life in HIV-infected patients on highly active antiretroviral therapy. AB - BACKGROUND: The overall health status and survival of HIV-infected patients has changed with the advent of highly active antiretroviral therapy (HAART). With this improved survival, there is a greater urgency to study quality-of-life issues. OBJECTIVE: Our objectives were to measure skin-related quality of life in a cohort of HIV-infected patients and to determine whether the use of highly active antiretroviral therapy is associated with improved skin-related quality of life. METHODS: We assembled a retrospective cohort of patients who were seen in our HIV-Dermatology Clinic at San Francisco General Hospital in June, July, or August of 1996. Eligible subjects were contacted by mail and asked (1) to complete a questionnaire (Skindex) and (2) to have a skin exam. Information on medication use and laboratory parameters was also collected. RESULTS: Of 107 eligible patients, 76 (71%) responded to the questionnaire; 60 patients were examined. Many patients had multiple skin conditions. For most diagnoses (except warts and onychomycosis), there were no consistent differences in Skindex scores of HIV-infected patients compared with scores of patients not known to be infected with HIV. Patients on HAART for longer duration had significantly lower Skindex scores (improved skin-related quality of life) compared with those on HAART for a shorter duration. CONCLUSION: HAART is associated with improved quality of life with regard to HIV-associated skin diseases. PMID- 11896419 TI - Herpetic folliculitis. AB - BACKGROUND: Although herpetic skin infection is very common, herpetic folliculitis is infrequently reported in the literature. It has varied presentations, some of which are clinically atypical requiring histopathological confirmation of follicular involvement. OBJECTIVE: We describe an otherwise healthy young adult male with extensive herpetic sycosis of the beard area, which is a variant of herpetic folliculitis. The diagnosis was confirmed by typical herpetic cytopathic changes in Tzanck smear and positive viral culture for HSV-1. METHOD: This article includes a case report and a literature review of herpetic (simplex and varicella/zoster) folliculitis. CONCLUSIONS: More cases of herpetic folliculitis should be reported to improve our understanding of this disease entity. Physicians should consider herpetic or other viral etiology in patients with folliculitis even if they were healthy, especially if they show resistance to antibacterial and antifungal therapy. PMID- 11896420 TI - Keratoelastoidosis marginalis. AB - BACKGROUND: Keratoelastoidosis marginalis is a rare disease that is a variant of solar elastosis. Long-term ultraviolet radiation exposure and chronic trauma secondary to manual labor are considered to be inciting factors. OBJECTIVE: To outline the clinical and histological features of this disorder. METHODS: We report two cases and review the literature. RESULTS: Both patients had persistence of their disease despite multiple topical treatment regimens. CONCLUSION: Keratoelastoidosis marginalis is a chronic and progressive disease that is difficult to treat. The differential includes focal acral hyperkeratosis and acrokeratoelastoidosis. PMID- 11896418 TI - An unusual case of cutaneous pancreatic fat necrosis. AB - BACKGROUND: Cutaneous pancreatic fat necrosis is a pathognomonic sign for pancreatic disease and usually presents as subcutaneous nodules in the pretibial region. OBJECTIVE: A case of cutaneous pancreatic fat necrosis is presented in which the clinical presentation of diffuse erythema was unusual. This disease is discussed and its possible etiologies are reviewed. METHODS: A MEDLINE search for cases of cutaneous pancreatic fat necrosis presenting as diffuse erythema without nodules was conducted. RESULTS: Diffuse erythema is an unusual presentation of cutaneous pancreatic fat necrosis. CONCLUSION: This may be the first case of cutaneous pancreatic fat necrosis presenting as diffuse erythema. PMID- 11896421 TI - Perianal basal cell carcinoma. AB - BACKGROUND: A case of basal cell carcinoma (BCC) of the perianal region is reported. This tumor is extremely rare in this location and behaves rather innocently. OBJECTIVE: Clinical and histopathologic characteristics of perianal BCC, as well as the choices of treatment, are outlined. CONCLUSION: The tumor should be histologically distinguished from basaloid carcinoma of the anus, which is much more aggressive and metastasizes early, thus requiring a different therapy. PMID- 11896422 TI - A listing of skin conditions exhibiting the koebner and pseudo-koebner phenomena with eliciting stimuli. AB - BACKGROUND: The Koebner phenomenon was first described in 1872. It occurs after a variety of traumatic insults. New examples of koebnerization are reported each decade. OBJECTIVE: We have compiled, for the first time in the dermatologic literature, an extensive list of the cutaneous diseases reported to demonstrate koebnerization, correlated in each case with the precise source of trauma. Cutaneous diseases reported to show a pseudo-Koebner phenomenon and the corresponding mechanism of trauma are also detailed. METHODS: A search and review of the English language literature on MEDLINE was made to identify cutaneous diseases exhibiting the Koebner phenomenon and the correlated mechanism of trauma. RESULTS: Results of our efforts are cataloged in table format. CONCLUSION: We present an extensive list of cutaneous diseases reported to exhibit the Koebner and pseudo-Koebner phenomena, correlating each entry with the mechanism of trauma. PMID- 11896423 TI - Is there an upper age limit for bone marrow transplantation? AB - Advances in hematopoietic cell transplantation have reduced the toxicity of both allogeneic and autologous transplantation. Decisions regarding the feasibility of transplantation should be individualized, and based upon physiological rather than chronological age. PMID- 11896424 TI - Mobilization of peripheral blood stem cells with high-dose cyclophosphamide or the DHAP regimen plus G-CSF in non-Hodgkin's lymphoma. AB - Our study analyzes the mobilization of hematopoietic stem cells after two chemotherapeutic regimens in non-Hodgkin's lymphoma (NHL) patients. The study included 72 patients with NHL (42 follicular and 30 large cells). The mean age was 37 years (range 17-60). Sixty-four patients (88.9%) had stage III-IV disease. Forty-eight patients (66.7%) had bone marrow involvement. Systemic B symptoms were present in 42 patients (58.3%). Mobilization chemotherapy regimens were randomly assigned as DHAP in 38 patients (52.7%) or cyclophosphamide (CPM) (5 g/m(2)) in 34 (47.2%) and the results of 132 procedures were analyzed. At the time of PBSC mobilization, 46 patients (63.9%) were considered to be responsive (complete remission, partial remission or sensitive relapse) and 26 (36.1%) not responsive (refractory relapse or refractory to therapy). Pre-apheresis CD34+ blood cell count and number of previous chemotherapy treatments were used to predict the total number of CD34+ cells in the apheresis product. The mobilizing regimens (CPM or DHAP) were similar in achieving the threshold CD34+ cell yield, for optimal engraftment. Since DHAP was very effective as salvage treatment, we suggest using DHAP as a mobilizing regimen in patients with active residual lymphoma at the time of stem cell collection. PMID- 11896425 TI - Allogeneic transplantation from HLA-matched sibling or partially HLA-mismatched related donors for primary refractory acute leukemia. AB - Allogeneic transplantation is successful in a minority of patients with primary refractory acute leukemia (PRAL). An HLA-matched sibling donor (MSD) is available only in 30-40% of the patients, whereas a partially mismatched related donor (PMRD) is available for most. We compared the outcome of 24 MSD (median age 24 years) and 19 PMRD (median age 34 years; P = 0.04) allograft recipients with PRAL. All MSD patients received non-T cell-depleted marrow whereas all PMRD patients received partially T cell-depleted marrow. All evaluable PMRD patients and 90% of the evaluable MSD patients attained CR. Six patients in each group with recurrent/persistent disease died. Ten PMRD (3-year probability 70%) and 14 MSD (3-year probability 63%) patients died of treatment-related causes. At the last follow-up, three PMRD (18-50 months; 3-year probability 14%) and four MSD (20-166 months; 3-year probability 20%) patients were alive and well. We conclude that allogeneic transplantation is a viable therapeutic option for PRAL. PMRD transplantation is a reasonable alternative in patients with no MSD, and results in similar outcome. In terms of identifying a donor and harvesting cells, a PMRD transplant is significantly quicker than an unrelated donor transplant - a point of great practical importance in the setting of failed induction chemotherapy where time is of the essence. PMID- 11896426 TI - Autologous stem cell transplantation for advanced acute myeloid leukemia. AB - We studied the efficacy of a two-step approach to autologous stem cell transplantation for patients with advanced acute myeloid leukemia. Step 1 consisted of consolidation chemotherapy using cytarabine 2000 mg/m(2) twice daily for 4 days plus etoposide 40 mg/kg by continuous infusion over the same 4 days. Peripheral blood stem cells were collected under granulocyte colony-stimulating factor (G-CSF) stimulation during recovery from this chemotherapy. Step 2, autologous stem cell transplantation, utilized the preparative regimen of oral busulfan 16 mg/kg followed by etoposide 60 mg/kg i.v. During step 1, there were no treatment-related deaths among 28 patients, but two patients did not proceed to transplantation because of failure of mobilization. A median CD34+ dose (x10(6)/kg) of 13.6 was collected. Of 26 patients undergoing autologous transplant, there was one treatment-related death and 12 relapses. With a median follow-up of 5.4 years, 5 year event-free survival (EFS) of all patients entered is 54%. The most important prognostic factor was cytogenetic changes. All seven patients with t(15,17) remained in long-term remission whereas EFS for other patients was 38%. We conclude that this two-step approach to autologous transplantation produces excellent stem cell yields, allows a high percentage of patients to receive the intended therapy, and provides effective treatment. PMID- 11896427 TI - Autotransplantation for advanced lymphoma and Hodgkin's disease followed by post transplant rituxan/GM-CSF or radiotherapy and consolidation chemotherapy. AB - Disease relapse occurs in 50% or more of patients who are autografted for relapsed or refractory lymphoma (NHL) or Hodgkin's disease (HD). The administration of non-cross-resistant therapies during the post-transplant phase could possibly control residual disease and delay or prevent its progression. To test this approach, 55 patients with relapsed/refractory or high-risk NHL or relapsed/refractory HD were enrolled in the following protocol: stem cell mobilization: cyclophosphamide (4.5 g/m(2)) + etoposide (2.0 g/m(2)) followed by GM-CSF or G-CSF; high-dose therapy: gemcitabine (1.0 g/m(2)) on day -5, BCNU (300 mg/m(2)) + gemcitabine (1.0 g/m(2)) on day -2, melphalan (140 mg/m(2)) on day -1, blood stem cell infusion on day 0; post-transplant immunotherapy (B cell NHL): rituxan (375 mg/m(2)) weekly for 4 weeks + GM-CSF (250 microg thrice weekly) (weeks 4-8); post-transplant involved-field radiotherapy (HD): 30-40 Gy to pre transplant areas of disease (weeks 4-8); post-transplant consolidation chemotherapy (all patients): dexamethasone (40 mg daily)/cyclophosphamide (300 mg/m(2)/day)/etoposide (30 mg/m(2)/day)/cisplatin (15 mg/m(2)/day) by continuous intravenous infusion for 4 days + gemcitabine (1.0 g/m(2), day 3) (months 3 + 9) alternating with dexamethasone/paclitaxel (135 mg/m(2))/cisplatin (75 mg/m(2)) (months 6 + 12). Of the 33 patients with B cell lymphoma, 14 had primary refractory disease (42%), 12 had relapsed disease (36%) and seven had high-risk disease in first CR (21%). For the entire group, the 2-year Kaplan-Meier event free survival (EFS) and overall survival (OS) were 30% and 35%, respectively, while six of 33 patients (18%) died before day 100 from transplant-related complications. The rituxan/GM-CSF phase was well-tolerated by the 26 patients who were treated and led to radiographic responses in seven patients; an eighth patient with a blastic variant of mantle-cell lymphoma had clearance of marrow involvement after rituxan/GM-CSF. Of the 22 patients with relapsed/refractory HD (21 patients) or high-risk T cell lymphoblastic lymphoma (one patient), the 2 year Kaplan-Meier EFS and OS were 70% and 85%, respectively, while two of 22 patients (9%) died before day 100 from transplant-related complications. Eight patients received involved field radiation and seven had radiographic responses within the treatment fields. A total of 72 courses of post-transplant consolidation chemotherapy were administered to 26 of the 55 total patients. Transient grade 3-4 myelosuppression was common and one patient died from neutropenic sepsis, but no patients required an infusion of backup stem cells. After adjustment for known prognostic factors, the EFS for the cohort of HD patients was significantly better than the EFS for an historical cohort of HD patients autografted after BEAC (BCNU/etoposide/cytarabine/cyclophosphamide) without consolidation chemotherapy (P = 0.015). In conclusion, post-transplant consolidation therapy is feasible and well-tolerated for patients autografted for aggressive NHL and HD and may be associated with improved progression-free survival particularly for patients with HD. PMID- 11896428 TI - Taurolidine: preclinical evaluation of a novel, highly selective, agent for bone marrow purging. AB - Taurolidine has been shown to have remarkable cytotoxic activity against selected human tumor cells at concentrations that spare normal cells. In this study we have extended this observation and assessed the ability of Taurolidine to purge tumor cells from chimeric mixtures of bone marrow (BM) and neoplastic cells. Normal murine BM and human leukemic (HL-60) or ovarian (PA-1) tumor cell lines were used as models. Exposure of tumor cells to 2.5 mM Taurolidine for 1 h resulted in the complete elimination of viable cells. In contrast, exposure of BM to 5 mMTaurolidine for 1 h reduced CFU-GM, BFU-E and CFU-GEEM colony formation by only 23.0%, 19.6% and 25.2%, respectively. Inhibition of long-term BM culture (LTBMC) growth following a 1 h exposure to 5 mM Taurolidine also was approximately 20% compared to untreated LTBMC. Finally, chimeric cultures were generated from BM and HL-60GR or PA-1GR cells (tumor cells transfected with the geneticin resistance gene). Exposure of these chimeric cultures to 5 mM Taurolidine for 1 h totally eliminated viable cancer cells while minimally reducing viable BM cells. This finding was confirmed by subsequent positive selection for surviving tumor cells with geneticin. These findings reveal that Taurolidine holds promise for use in BM purging. PMID- 11896429 TI - Respiratory syncytial virus infection following hematopoietic stem cell transplantation. AB - Respiratory syncytial virus, one of the most common causes of respiratory infections in immunocompetent individuals, is frequently spread to recipients of HSCT by family members, other patients, and health care workers. In immunosuppressed individuals, progression from upper respiratory tract disease to pneumonia is common, and usually fatal if left untreated. We performed a retrospective analysis of RSV infections in recipients of autologous or allogeneic transplants. The incidence of RSV following allogeneic or autologous HSCT was 5.7% and 1.5%, respectively. Of the 58 patients with an RSV infection, 16 of 21 patients identified within the first post-transplant month, developed pneumonia. Seventy-two percent of patients received aerosolized ribavirin and/or RSV-IGIV, including 23 of 25 patients diagnosed with RSV pneumonia. In this aggressively treated patient population, three patients died of RSV disease, each following an unrelated HSCT. PMID- 11896432 TI - The role of biomedical and psychosocial factors for the prediction of pain and distress in patients undergoing high-dose therapy and BMT/PBSCT. AB - Recent research has shown that cancer patients undergoing bone marrow transplantation (BMT) experience moderate to severe mouth pain due to treatment related mucositis in spite of morphine therapy. Treatment-related emotional distress in BMT patients is also described widely. This study examined several biomedical, psychological and social variables as possible predictors for the intensity of treatment-related mouth pain and anxious mood in 63 cancer patients undergoing BMT or stem cell transplantation (SCT) within a prospective longitudinal design. Biomedical predictors included biomedical risk, mucositis, the mode of transplantation, total body irradiation, age and gender. Psychological predictors were depression (BDI), BMT-related distress, chronic stress and resources in everyday life (KISS), pain-related coping behaviour (KPI 17) and social support (ISSS). Among the social variables we evaluated education, being married and the living situation. Criteria variables were the intensity of mouth pain and anxious mood which were assessed daily by numeric self-rating scales for 24 days after transplantation. Results of stepwise multiple regressions indicated that psychological and social variables were important predictors of mouth pain, besides biomedical variables. Whereas the biomedical variables revealed the most predictive power during the second week after BMT, psychological predictors were more important during the early and late phases of the treatment. Daily anxious mood was best predicted by psychological and social variables. Among the biomedical variables mucositis was most strongly related to mouth pain besides mode of transplantation, risk, TBI and age. Among the psychological variables BMT-related distress was the most important predictor, with resources in private life or at work and pain-related coping modes as further significant predictors. These results imply that relevant predictors should be assessed as high risk factors for an increased vulnerability for treatment-related side-effects before treatment starts indicating an additional offer of psychological treatment in high risk patients. PMID- 11896431 TI - Increased incidence of EBV-associated lymphoproliferative disorders after allogeneic stem cell transplantation from matched unrelated donors due to a change of T cell depletion technique. AB - Here, the influence of T vs T and B cell depletion on the incidence of EBV associated lymphoproliferative disorder (EBV-LPD) after bone marrow transplantation (BMT) from a matched unrelated donor (MUD) is analyzed. From 1982 to 1997 the soy bean agglutinin/sheep red blood cell (SBA/SRBC) method was used for T cell depletion. This technique is well established, but the use of SRBC has a risk of transmitting prions or viruses. Therefore, a new T cell depletion method was introduced, using CD2 and CD3 monoclonal antibodies (CD2/3 method) instead of SRBC. Unfortunately, this led to an unexpected high number of EBV-LPDs in patients receiving transplants from MUDs. SBA depletion was reintroduced and combined with the CD2/3 method (SBA/CD2/3) in this patient population, later replaced by B cell-specific (CD19 and CD22) antibodies (CD3/19/22 method). The number of T (x 10(5)/kg) and B (x 10(5)/kg) cells in the graft was 1.5 +/- 0.8 and 2 +/- 1 (T/B ratio 0.75), 2.2 +/- 2.0 and 41 +/- 21 (ratio 0.055), 5.0 +/- 0.0 and 2 +/- 1 (ratio 2.5), 2.5 +/- 1.2 and 10 +/- 6 (ratio 0.25) using the SBA/SRBC, CD2/3, SBA/CD2/3 and CD3/19/22 techniques, respectively. When B cell depletion was performed (SBA/SRBC, SBA/CD2/3, CD3/19/22) four out of 31 patients (13%) receiving a BMT from a MUD developed an EBV-LPD. Without B cell depletion (CD2/3) this occurred in five out of seven patients (71%) (P < 0.05). A T/B cell ratio in the graft of > or = 0.25 seems sufficient to significantly reduce the incidence of EBV-LPD after BMT from MUDs. PMID- 11896430 TI - Portal vein thrombosis after hematopoietic cell transplantation: frequency, treatment and outcome. AB - Patients who develop veno-occlusive disease (VOD) of the liver may have low plasma levels of the natural anticoagulants protein C and antithrombin III, but large vessel thromboses are not commonly reported in these patients. We reviewed the records of 1847 consecutive patients for evidence of portal vein thrombosis. Eight patients (0.4%) developed portal vein thrombosis (PVT) at a median of day +28 (range 3-58). All patients had clinical evidence of VOD with ascites, a median total serum bilirubin 11.9 mg/dl, and median weight gain from baseline of 7.9%. Median plasma levels of antithrombin III and protein C were low (36% and 21%, respectively). Four patients with PVT died of severe VOD and multi-organ failure, but PVT did not contribute to death. We conclude that PVT is a rare complication of hematopoietic cell transplant and is associated with hepatic VOD. We speculate that PVT resulted from diminished portal venous flow (related to hepatic sinusoidal obstruction to blood flow) and a hypercoagulable state (related to low circulating antithrombin III and protein C levels). Prognosis depended on the severity of the underlying VOD and not PVT per se, suggesting that treatments directed solely toward dissolution of portal vein thrombi should be used with caution in this setting. PMID- 11896433 TI - Radiologically guided fine needle lung biopsies in the evaluation of focal pulmonary lesions in allogeneic stem cell transplant recipients. AB - Lung problems are common in allogeneic stem cell transplant (SCT) recipients. To evaluate the feasibility and diagnostic yield of radiologically guided fine needle lung biopsy (FNLB) in allogeneic SCT recipients with focal pulmonary lesions, a retrospective analysis was carried out. Between 1989 and 1998, radiologists performed a total of 30 FNLBs in 21 allogeneic SCT recipients, guided either by ultrasound (n = 17) or computed tomography (n = 13). The median time from SCT to the first FNLB was 131 days (20-343 days). Prophylactic platelet transfusions were given in 19 procedures (66%). The complications of FNLB included clinically insignificant pneumothorax in four procedures (13%) and self limiting haemoptysis in one case (3%). The first FNLB was suggestive of invasive pulmonary aspergillosis (IPA) in five patients (24%). Additional clinically useful findings of FNLB included Pseudomonas (two patients) and Nocardia (one patient). The final diagnosis of pulmonary lesions was IPA in 14 patients, immunological lung problems in four patients and other in three patients. Radiologically guided FNLB is feasible in allogeneic SCT recipients and has a low complication rate. The diagnostic yield is high especially for IPA. PMID- 11896434 TI - Progressive interstitial fibrosis of the lung in sclerodermoid chronic graft versus-host disease. AB - Sclerodermoid chronic graft-versus-host disease (sGVHD) is a well-known complication in patients with a long history of chronic GVHD. Pulmonary involvement in chronic GVHD presents typically as bronchiolitis obliterans (BO). Pulmonary fibrosis after allogeneic hematopoietic stem cell transplantation (HSCT) is presumed to be caused by the long-term toxicity of the conditioning regimen or the result of lung injury elicited predominantly by viral infections or GVHD. We present two patients with late onset pulmonary fibrosis associated with moderate sGVHD of the skin after HSCT. At the initial diagnosis of chronic GVHD both patients presented with symptoms of interstitial pneumonia. Years later both patients developed moderate to severe interstitial pulmonary fibrosis in association with sGVHD. One patient showed additional clinical and histological signs of BO. While one patient responded to increased immunosuppression including total nodal irradiation (1 Gy), the other patient died due to complications related to pulmonary fibrosis. PMID- 11896435 TI - Lamivudine treatment for reverse seroconversion of hepatitis B 4 years after allogeneic bone marrow transplantation. AB - Reverse seroconversion of hepatitis B virus (HBV) after allogeneic BMT is rare. We present a case of HBV reactivation late after allogeneic BMT which responded well to lamivudine therapy. A 35-year-old woman with CML received an allogeneic BMT. Before BMT, the patient had immunity to HBV, with serum antibodies against hepatitis B surface antigen (HBsAb), and the donor was completely negative for HBV. Four years after BMT, acute hepatitis occurred with a detectable level of HBV-DNA. Lamivudine rapidly reduced transaminase and bilirubin levels, and serum HBV-DNA decreased to negative. Retrospective analysis revealed that there had been a gradual decrease in serum HBsAb titers after BMT. Administration of lamivudine immediately after HBV replication may be more effective than vaccination of hepatitis B surface antigen-negative donors before BMT. PMID- 11896436 TI - Anti-CD20 monoclonal antibody therapy for autoimmune hemolytic anemia following T cell-depleted, haplo-identical stem cell transplantation. PMID- 11896437 TI - Wild-type p53 induced sensitization of mutant p53 TNF-resistant cells: role of caspase-8 and mitochondria. AB - In the present study, we have investigated the mechanisms by which the restoration of wild-type (wt) p53 functions in p53 mutant cells increases their susceptibility to the cytotoxic action of tumor necrosis factor (TNF). Our data indicate that the resistance of p53-mutated cl.1001 cells to TNF-induced cell death was not due to a defect in the expression of TRADD and FADD, yet correlated with a reduced caspase-8 activation as well as a deficient mitochondrial membrane permeabilization. Moreover, cl.1001 cells failed to translocate the mitochondrial AIF and cytochrome c to the nucleus and to the cytosol, respectively, in response to TNF. Sensitization of these cells, following infection with a recombinant adenovirus encoding wtp53, to TNF-induced cytotoxicity resulted in the restoration of caspase-8 cleavage and the reestablishment of mitochondrial signs of apoptosis. These findings suggest that the cross-talk between p53 and TNF induced cell death depends on mitochondria and that the combination of TNF and Adwtp53 may be a potential strategy to sensitize mutant p53 TNF-resistant tumors to the cytotoxic action of this cytokine. PMID- 11896438 TI - Intra-arterial delivery of p53-containing adenoviral vector into experimental brain tumors. AB - Human tumor xenografts established in athymic rat brains were used to determine the feasibility of intravascular delivery of tumor suppressor genes to brain tumors. Both tumor size and number were compared to characterize the effect of tumor burden on tumor transduction efficacy by a control LacZ-containing adenoviral vector. Experiments with tumors grown in vivo for either 3, 5, or 7 days demonstrated that 5-day-old tumors provided the best target for vector infection and transgene expression by this mode of administration. Intra-arterial mannitol facilitated transduction efficiency. Tumor burden did not seem to affect transduction, while tumor location appeared to be an important factor. Based on these results, intra-arterial infusion of a p53-containing adenoviral vector was carried out and resulted in significant retardation of brain tumor growth 3 days after administration. Effects at longer time points were not as significant. These findings indicate that intra-arterial administration of adenoviral vectors containing p53 is efficient and can result in changes in tumor size, but that long-term control of tumor growth may require multiple adenoviral treatments. PMID- 11896439 TI - Enhanced antitumor effect and reduced vector dissemination with fiber-modified adenovirus vectors expressing herpes simplex virus thymidine kinase. AB - There are at least two hurdles confronting the use of the adenovirus (Ad) mediated herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) system for the treatment of cancer. One is inefficient Ad vector-mediated gene transfer into tumor cells lacking the primary receptor, i.e., the coxsackievirus and adenovirus receptor (CAR). The other is hepatotoxicity due to unwanted vector spread into the liver, even when Ad vectors are injected intratumorally. Herein, we present an attractive strategy for overcoming such limitations based on use of a fiber-modified Ad vector containing an RGD peptide motif in the fiber knob. HSVtk-expressing Ad vectors containing mutant fiber (AdRGD-tk) or wild-type fiber (Ad-tk) were injected intratumorally into CAR-negative B16 melanoma cells inoculated into mice, after which GCV was injected intraperitoneally for 10 days. AdRGD-tk showed approximately 25 times more antitumor activity than Ad-tk. Histopathological studies suggested that liver damage in mice injected with AdRGD tk was significantly lower than that in mice injected with Ad-tk. Intratumoral administration of luciferase-expressing Ad vectors containing the mutant fiber (AdRGD-L2) resulted in nearly 40 times more luciferase production in the tumor, but 8 times less production in the liver than the conventional Ad vectors (Ad L2). These results indicate that combination of fiber-modified vectors and a HSVtk/GCV system is a potentially useful and safe approach for the treatment of tumors lacking CAR expression, and that fiber-modified vectors could be of great utility for gene therapy and gene transfer experiments. PMID- 11896440 TI - Comparative assessment of TCRBV diversity in T lymphocytes present in blood, metastatic lesions, and DTH sites of two melanoma patients vaccinated with an IL 7 gene-modified autologous tumor cell vaccine. AB - A phase I clinical trial using autologous, IL-7 gene-modified tumor cells in patients with disseminated melanoma has been recently completed. Although no major clinical responses were observed, increased antitumor cytotoxicity was measured in postvaccine peripheral blood lymphocytes in a subset of treated patients. To analyze the in situ immune response, the T cell receptor beta-chain variable region (BV) repertoire of T cells infiltrating postvaccine lesions was studied in two patients, and compared with that of T cells present in prevaccine ones, in peripheral blood lymphocytes, and, in one patient, in delayed type hypersensitivity (DTH) sites of autologous melanoma inoculum. A relative expansion of T cells expressing few BVs was observed in all postvaccine metastases, and their intratumoral presence was confirmed by immunohistochemistry. Length pattern analysis of the complementarity determining region 3 (CDR3) indicated that the repertoire of T cells expressing some of these BVs was heterogeneous. At difference, CDR3, beta-chain joining region usage, and sequence analysis enabled us to demonstrate, within a T-cell subpopulation commonly expanded at DTH sites and at the postvaccine lesion of patient 1, that both DTH sites contained identical dominant T-cell clonotypes. One of them was also expressed at increased relative frequency in the postvaccine lesion compared to prevaccine specimens. These results provide evidence for immunological changes, including in situ clonally expanded T cells, in metastases of patients vaccinated with IL-7 gene-transduced cells. PMID- 11896441 TI - Clinical trial of E1B-deleted adenovirus (dl1520) gene therapy for hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. The prognosis of HCC is poor and current therapies are largely ineffective. Genetic abnormalities are commonly seen in HCC tumors particularly with inactivation of the p53 tumor suppressor. Gene therapy with E1B-deleted (dl1520) adenovirus could be of therapeutic value as it offers the potential of tumor growth control in patients with p53 mutation. Ten patients with posthepatitis cirrhosis and histologically proven HCC were enrolled into an open label, randomized prospective study. Randomization was to receive either percutaneous ethanol injection (control group) or dl1520. Toxicity and complications in the ethanol group were pain and fever, whereas in the gene therapy group complications were minimal. Grade I-II toxicity fever, stable performance status, and no significant rise in liver enzymes were observed in patients treated with dl1520. Analysis of patients' response to treatment in the gene therapy group showed one patient with a partial response and four patients with progressive disease. In the ethanol treated group two patients had stable disease and three patients showed disease progression. In conclusion, this study showed that the adenovirus was well tolerated, but did not seem to offer significant tumor control. Although only a small number of patients were treated here it appears that more effective vectors are needed to achieve a useful clinical impact. PMID- 11896443 TI - Gene transfer into human prostate adenocarcinoma cells with an adenoviral vector: Hyperthermia enhances a double suicide gene expression, cytotoxicity and radiotoxicity. AB - We have previously developed a recombinant adenovirus containing a fusion gene of Escherichia coli cytosine deaminase (CD) and herpes simplex virus type 1 thymidine kinase (HSV-1 TK) controlled by a cytomegalovirus (CMV) enhancer promoter. This replication-incompetent adenovirus effectively transduced the CD TK gene into human prostate adenocarcinoma DU-145 or PC-3 cells. Interestingly, heat shock at 41 degrees C for 4 hours elevated the level of CD-TK by approximately 5- to 20-fold at a multiplicity of infection (MOI) of 1. Heat enhanced expression of CD-TK promoted cytotoxicity by 23-, 9-, or 47-fold in the presence of 50 microg/mL ganciclovir (GCV), 500 microg/mL 5-fluorocytosine (5 FC), or 50 microg/mL GCV+500 microg/mL 5-FC, respectively, at an MOI of 1. Moreover, there was an increase in radiosensitivity when adenovirus-infected cells were heated at 41 degrees C for 4 hours followed by irradiation in the presence of the prodrugs. Virus+heat+1 microg/mL GCV treatment increased radiosensitivity by a dose-modifying factor (DMF) of 2.2, whereas virus+heat+10 microg/mL 5-FC exposure resulted in a DMF of 2.3. Radiosensitization was clearly enhanced as a result of combined prodrug exposure (DMF=4.4). Our results suggest that the efficiency in expression of suicide genes from an adenoviral vector used for cytotoxic anticancer therapy could be improved by combining heat treatment with radiation therapy. PMID- 11896442 TI - Eradication of osteosarcoma lung metastases following intranasal interleukin-12 gene therapy using a nonviral polyethylenimine vector. AB - The use of adenoviral vectors for therapeutic delivery of genes via pulmonary application poses several problems in terms of immune responses. The purpose of this study was to determine whether polyethylenimine (PEI), a polycationic DNA carrier, can be used to deliver the IL-12 gene into the lungs of mice having microscopic osteosarcoma (OS) lung metastases. Incubation of SAOS-LM6 cells in vitro with PEI containing the murine IL-12 (mIL-12) gene (PEI:IL-12) resulted in expression of both the p35 and p40 subunits of IL-12 mRNA and production of mIL 12 protein. Using our newly developed OS nude mouse model, we demonstrated that treatment of mice using intranasal PEI:IL-12 resulted in significant IL-12 mRNA expression in the lung but not the liver. Furthermore, plasma IL-12 was undetectable after up to 4 weeks of intranasal PEI:IL-12 therapy given twice weekly. No IL-12 expression was seen following intranasal PEI therapy alone. The number of lung metastases in animals that received intranasal PEI:IL-12 twice weekly for 4 weeks starting 6 weeks after tumor inoculation was significantly decreased (median, 11; range, 0-47) compared with those that received PEI alone (median, 89; range, 2 to >200; P=.012). Also, the size of the nodules was significantly smaller in the PEI:IL-12-treated animals, with 90% measuring < or =0.5 mm in diameter compared with 56% in the PEI-alone group. Animals that received PEI alone also had numerous large nodules (3-6 mm) throughout the lungs. Intranasal therapy is a noninvasive way to administer agents and has the advantage of targeting the pulmonary region, resulting in higher concentrations in the tumor area. Additionally, delivery of IL-12 to the lung via the airway using PEI may avoid systemic toxicity. Because OS metastasizes almost exclusively to the lung, this may be a novel approach to the treatment of pulmonary OS metastases. PMID- 11896444 TI - Autocrine costimulation: tumor-specific CD28-mediated costimulation of T cells by in situ production of a bifunctional B7-anti-CEA diabody fusion protein. AB - T cells require two distinct signals for optimal activation, an antigen-specific signal, provided by engagement of the T-cell receptor (TCR) and a second costimulatory signal mediated by engagement of CD28 with members of the B7 family. Although infiltrating T cells are present in many malignancies, they appear to be mostly anergic and do not attack the tumor, presumably because of the absence of activation and/or costimulatory signals. Here we describe a novel strategy for the in situ activation of tumor-specific T cells. We genetically modified T cells to secrete a bifunctional fusion protein, comprising the extracellular portion of B7-1 fused to an anticarcinoembryonic antigen (CEA) diabody. In coculture with CEA(+) tumor cells autocrine and paracrine secretion of B7-alphaCEA provided a potent tumor-specific costimulatory signal to T cells in combination with a recombinant alphaCEAxalphaCD3 bispecific diabody. B7 alphaCEA was also found to strongly enhance survival and tumor-specific activation of T cells expressing an anti-CEA TCRzeta-based chimeric immune receptor (CIR) both when expressed in cis by the T cells themselves as well as in trans, when added to the culture medium. In the absence of costimulatory signals provided by the tumor, our strategy allows T cells to "arm themselves" by the production of tumor-specific costimulatory proteins. Sustained in situ production of such molecules, like the B7-diabody fusion protein may create a favorable local environment for the activation and proliferation of tumor-reactive T cells and increase the tumoricidal activity of immunotherapeutic approaches targeting the TCR pathway. PMID- 11896445 TI - In vivo induction of antitumor immunity and protection against tumor growth by injection of CD154-expressing tumor cells. AB - As they should enhance tumor-specific antigen presentation by dendritic cells, tumor cell lines genetically modified to express CD154 molecules have been used in an attempt to induce protective antitumor immunity. Two murine models were used: the major histocompatibility complex (MHC) class I negative melanoma B16F10 and the MHC class I positive mammary adenocarcinoma TS/A. CD154 or mock transfected B16F10 or TS/A cells were injected subcutaneously into H-2-compatible B6D2 mice. CD154 expression by tumor cells induced a complete rejection (in the TS/A model) or a striking reduction (in the B16F10 model) of modified tumors growth, but also a significant protection against the growth of mock tumor cells injected simultaneously, either mixed with the CD154-expressing tumor cells, or in the other flank of mice. Thirty days after CD154-expressing tumor rejection, splenic lymphocytes from surviving tumor-free mice were able to inhibit tumor proliferation in vitro and significant amounts of IFN-gamma were detected in the sera of these mice. Growth kinetics of mock and CD154-expressing tumors in immunocompetent versus nude mice suggest that T lymphocytes and natural killer cells responses are implicated in this antitumor immunity. The injection of CD154 expressing tumor cell induced an antitumor protective response, both locally and distant from the injection site. The effect was most pronounced in MHC class I expressing TS/A tumor model. PMID- 11896446 TI - Immunotherapy of metastatic melanoma by intratumoral injections of Vero cells producing human IL-2: phase II randomized study comparing two dose levels. AB - BACKGROUND: Systemic IL-2 has shown some activity in metastatic melanoma, but its use is severely limited by toxicity. TG2001 is a product in which the human IL-2 cDNA was incorporated into the genome of Vero cells, a monkey fibroblast cell line. The goal of this intratumorally applied therapy was to create an antitumor immune response stimulated by xeno-antigens and local production of IL-2 in the close vicinity of tumor-specific antigens. TG2001 was reported to have a good safety profile in two previous dose-escalating phase I studies performed in 18 patients with various solid tumors, with encouraging clinical responses in three patients. The objectives of this study were to evaluate the tolerance and incidence of tumor regression in patients with metastatic melanoma, following repeated administration of Vero-IL-2 cells. PATIENTS AND METHODS: This was on open-label, randomized phase II study comparing two doses of Vero-IL-2, 5x10(5) and 5x10(6) cells. Twenty-eight patients with metastatic melanoma were enrolled in the study, 14 in each treatment group. Patients received TG2001 by intratumoral injection on days 1, 3, and 5 every 4 weeks for four cycles, and every 8 weeks thereafter, until evidence of progressive disease (PD). Criteria for patient selection included histologically proven metastatic melanoma, with one tumor accessible for product administration, and at least another tumor site for response assessment. Evaluation included tumor measurements, humoral and T cell-mediated local and systemic immune response, humoral response to Vero cells, adverse events and standard laboratory parameters. RESULTS: None of the patients achieved a confirmed objective response. Stable disease (SD) was seen in six (43%) and eight patients (57%) at the 5x10(5) and the 5x10(6) dose level, respectively. Two patients, one in each group, died during the study (i.e., within 1 month after the last injection) due to PD. Three patients exhibited antibody responses to Vero cells. T-cell immunity, serum cytokine levels and cytokine mRNA expression in tumor biopsies did not show meaningful alterations after therapy, except for a trend toward an increase in intratumoral TH2 cytokine (IL-4 and/or IL-10) levels. The study drug was well tolerated at both dose levels and side effects mainly consisted of injection site pain and erythema, and pyrexia. CONCLUSION: The intratumoral administration of TG2001 was generally well tolerated in patients with metastatic melanoma, and transient disease stabilization was observed in 50% of patients. PMID- 11896448 TI - Methotrexate selection of long-term culture initiating cells following transduction of CD34(+) cells with a retrovirus containing a mutated human dihydrofolate reductase gene. AB - A limitation of successful stem cell gene transfer to hematopoietic stem cells is low transduction efficiency. To overcome this hurdle and develop a gene transfer strategy that might be clinically feasible, retroviral vectors containing a drug resistance gene were utilized to transduce human CD34(+)-enriched cells and select gene-modified cells by drug administration. We constructed a high-titer retroviral vector containing a fusion gene (F/S-EGFP) consisting of a mutated dihydrofolate reductase (DHFR) (Leu22-->Phe22, Phe31-->Ser31; F/S) gene and enhanced green fluorescent protein (EGFP) cDNA. To test whether the fusion gene could function as a selectable marker, transduced CD34(+) cells were assayed in long-term stromal co-cultures with and without addition of methotrexate (MTX). Without MTX exposure, the vector-transduced CD34(+) cells generated 22-50% EGFP(+) cobblestone area forming cells (CAFC) at week 5. By contrast, the vector transduced cells cultured with MTX produced 96-100% EGFP(+) CAFC in four separate experiments. These are the first investigations to demonstrate selection for transduced long-term culture initiating cells using MTX. The DHFR/MTX system holds promise for improving selection of gene-transduced hematopoietic progenitor cells in vivo. PMID- 11896447 TI - Effectiveness of insulin-like growth factor I receptor antisense strategy against Ewing's sarcoma cells. AB - The insulin-like growth factor I receptor (IGF-IR) plays an essential role in the establishment and maintenance of transformed phenotype of Ewing's sarcoma (ES) cells, and interference with the IGF-IR pathways by a neutralizing antibody causes reversal of the malignant potential of this neoplasm. In this paper, we stably transfected an IGF-IR antisense mRNA expression plasmid in an ES cell line to determine the effectiveness of antisense strategies against the in vitro and in vivo growth of ES cells. Doxorubicin sensitivity of TC-71 cells expressing antisense targeted to IGF-IR mRNA was also examined. Cells carrying antisense IGF IR had a reduced expression of the receptor, a modest decrease in cell proliferation, a significant increase in anoikis-induced apoptosis, and a severely reduced ability to form colonies in soft agar. Moreover, TC/AS cells showed a marked reduction in their motility. In vivo, when cells carrying antisense IGF-IR were injected subcutaneously in nude mice, tumor formation was delayed and survival increased. Metastatic ability of ES cells was also significantly reduced. Furthermore, TC/AS clones showed a significantly higher sensitivity to doxorubicin - a major drug in the treatment of ES. These results indicate that inhibiting IGF-IR by antisense strategies may be relevant to the clinical treatment of ES patients by reducing the malignant potential of these cells and enhancing the effectiveness of chemotherapy. PMID- 11896450 TI - X-linked Coffin-Lowry syndrome (CLS, MIM 303600, RPS6KA3 gene, protein product known under various names: pp90(rsk2), RSK2, ISPK, MAPKAP1). AB - The Coffin-Lowry syndrome (CLS) is a syndromic form of X-linked mental retardation characterised in male patients by psychomotor and growth retardation, and various skeletal anomalies. CLS is caused by mutations in a gene located in Xp22.2 and encoding RSK2, a growth-factor regulated protein kinase. Mutations are extremely heterogeneous and lead to premature termination of translation and/or to loss of phosphotransferase activity. No correlation between the type and location of mutation and the clinical phenotype is evident. However, in one family (MRX19), a missense mutation was associated solely with mild mental retardation and no other clinical feature. Screening for RSK2 mutations is essential in most cases to confirm the diagnosis as well as for genetic counseling. PMID- 11896451 TI - DNA methylation, imprinting and cancer. AB - It is well known that a variety of genetic changes influence the development and progression of cancer. These changes may result from inherited or spontaneous mutations that are not corrected by repair mechanisms prior to DNA replication. It is increasingly clear that so called epigenetic effects that do not affect the primary sequence of the genome also play an important role in tumorigenesis. This was supported initially by observations that cancer genomes undergo changes in their methylation state and that control of parental allele-specific methylation and expression of imprinted loci is lost in several cancers. Many loci acquiring aberrant methylation in cancers have since been identified and shown to be silenced by DNA methylation. In many cases, this mechanism of silencing inactivates tumour suppressors as effectively as frank mutation and is one of the cancer-predisposing hits described in Knudson's two hit hypothesis. In contrast to mutations which are essentially irreversible, methylation changes are reversible, raising the possibility of developing therapeutics based on restoring the normal methylation state to cancer-associated genes. Development of such therapeutics will require identifying loci undergoing methylation changes in cancer, understanding how their methylation influences tumorigenesis and identifying the mechanisms regulating the methylation state of the genome. The purpose of this review is to summarise what is known about these issues. PMID- 11896452 TI - Physical and transcriptional map of the critical region for keratolytic winter erythema (KWE) on chromosome 8p22-p23 between D8S550 and D8S1759. AB - Keratolytic winter erythema is an autosomal dominant skin disorder characterised by erythema, hyperkeratosis, and peeling of the skin of the palms and soles, especially during winter. The keratolytic winter erythema locus has been mapped to human chromosome 8p22-p23. This chromosomal region has also been associated with frequent loss of heterozygosity in different types of cancer. To identify positional candidate genes for keratolytic winter erythema, a BAC contig located between the markers at D8S550 and D8S1695 was constructed and sequenced. It could be extended to D8S1759 by a partially sequenced BAC clone identified by database searches. In the 634 404 bp contig 13 new polymorphic microsatellite loci and 46 single nucleotide and insertion/deletion polymorphisms were identified. Twelve transcripts were identified between D8S550 and D8S1759 by exon trapping, cDNA selection, and sequence analyses. They were localised on the genomic sequence, their exon/intron structure was determined, and their expression analysed by RT PCR. Only one of the transcripts corresponds to a known gene, encoding B lymphocyte specific tyrosine kinase, BLK. A putative novel myotubularin-related protein gene (MTMR8), a potential human homologue of the mouse acyl-malonyl condensing enzyme gene (Amac1), and two transcripts showing similarities to the mouse L-threonine 3-dehydrogenase gene and the human SEC oncogene, respectively, were identified. The remaining seven transcripts did not show similarities to known genes. There were no potentially pathogenic mutations identified in any of these transcripts in keratolytic winter erythema patients. PMID- 11896453 TI - Human chromosome 15q11-q14 regions of rearrangements contain clusters of LCR15 duplicons. AB - Six breakpoint regions for rearrangements of human chromosome 15q11-q14 have been described. These rearrangements involve deletions found in approximately 70% of Prader-Willi or Angelman's syndrome patients (PWS, AS), duplications detected in some cases of autism, triplications and inverted duplications. HERC2-containing (HEct domain and RCc1 domain protein 2) segmental duplications or duplicons are present at two of these breakpoints (BP2 and BP3) mainly associated with deletions. We show here that clusters containing several copies of the human chromosome 15 low-copy repeat (LCR15) duplicon are located at each of the six described 15q11-q14 BPs. In addition, our results suggest the existence of breakpoints for large 15q11-q13 deletions in a proximal duplicon-containing clone. The study reveals that HERC2-containing duplicons (estimated on 50-400 kb) and LCR15 duplicons ( approximately 15 kb on 15q11-q14) share the golgin-like protein (GLP) genomic sequence. Through the analysis of a human BAC library and public databases we have identified 36 LCR15 related sequences in the human genome, most (27) mapping to chromosome 15q and being transcribed. LCR15 analysis in non-human primates and age-sequence divergences support a recent origin of this family of segmental duplications through human speciation. PMID- 11896454 TI - Characterisation of the human voltage-gated potassium channel gene, KCNA7, a candidate gene for inherited cardiac disorders, and its exclusion as cause of progressive familial heart block I (PFHBI). AB - Mutations in genes encoding cardiac ion channels and their subunits are responsible for several genetic cardiac disorders. We characterised the human gene KCNA7, encoding the voltage-gated potassium channel Kv1.7 and compared its coding sequence with that of the mouse orthologue, kcna7. Both genes are encoded by two exons separated by a conserved intron, unlike all the other Kv1-family genes that contain intronless coding regions. KCNA7 and kcna7 encode proteins of 456 amino acid residues that share >95% sequence identity, and the mouse channel has biophysical and pharmacological properties closely resembling the ultra rapidly activating delayed rectifier (I(Kur)) in cardiac tissue. Using reverse transcriptase-PCR, KCNA7 mRNA was detected in adult human heart. We determined that KCNA7 resides on chromosome 19q13.3 in a region that also contains the progressive familial heart block I (PFHBI) locus. Direct sequencing of KCNA7's coding sequence in PFHB1-affected individuals revealed no pathogenic sequence changes, but two single nucleotide polymorphisms detected in exon 2 result in amino acid substitutions. These results provide evidence for the exclusion of this candidate as the PFHB1-causative gene, although mutations in regulatory and non-coding regions cannot be excluded. As ion channel-encoding genes have been implicated in a growing number of genetic conditions, the data presented may facilitate further analysis of the role of KCNA7 and its product in the heart. PMID- 11896455 TI - Lack of expression of XIST from a small ring X chromosome containing the XIST locus in a girl with short stature, facial dysmorphism and developmental delay. AB - A 46,X,r(X) karyotype was found in a three and a half year old girl with short stature, facial dysmorphism and developmental delay. The clinical findings were consistent with the phenotype described in a limited number of patients with small ring X chromosomes lacking the XIST locus, a critical player in the process of X chromosome inactivation. Surprisingly, in our patient, fluorescent in situ hybridisation demonstrated that the XIST locus was present on the ring X. However, expression studies showed that there was no XIST transcript in peripheral blood cells, suggesting that the ring X had not been inactivated. This was confirmed by the demonstration that both of the patient's alleles for the androgen receptor gene were unmethylated, and that both of the patient's ZXDA alleles were expressed. The active nature of the ring X would presumably result in overexpression of genes that may account for the developmental delay observed for the patient. Using polymorphic markers along the X chromosome, the ring X was determined to be of paternal origin with one breakpoint in the long arm between DXS8037 and XIST and one in the short arm in Xp11.2 between DXS1126 and DXS991. To attempt to determine why the XIST gene failed to be expressed, the promoter region was sequenced and found to have a base change at the same location as a variant previously associated with nonrandom X chromosome inactivation. This mutation was not seen in over one hundred normal X chromosomes examined; however, it was observed in the paternal grandmother who did not show substantial skewing of X chromosome inactivation. PMID- 11896456 TI - The transcriptional map of the common eliminated region 1 (C3CER1) in 3p21.3. AB - Occurrence of chromosome 3p deletions in a large number of human tumours suggests the existence of uncharted tumour suppressor gene(s). We previously applied a functional assay, named the Elimination test (Et), for the identification of regions containing tumour growth antagonising genes. This resulted in the definition of chromosome 3 common eliminated region 1 (C3CER1) on 3p21.3, which is regularly eliminated from SCID-derived tumours. Systematic genomic sequencing of 11 PAC clones, combined with comparisons of genomic sequence against EST databases and PCR-based cloning of cDNA sequences allowed us to assemble a comprehensive transcriptional map of 1.4 Mb that includes 19 active genes and three processed pseudogenes. We report four novel genes: FYVE and coiled-coil domain containing 1 (FYCO1), transmembrane protein 7 (TMEM7), leucine-rich repeat containing 2 (LRRC2) and leucine zipper protein 3 (LUZP3). A striking feature of C3CER1 is a presence of a cluster of eight chemokine receptor genes. Based on a new analysis of the microcell hybrid-derived panel of SCID tumours we also redefined the centromeric border of the C3CER1. It is now located within LRRC2 gene, which is a relative of RSP-1 (Ras Suppressor Protein 1). The detailed knowledge of gene content in C3CER1 is a prerequisite for functional analysis of these genes and understanding of their possible role in tumorigenesis. PMID- 11896458 TI - A large deletion including most of GJB6 in recessive non syndromic deafness: a digenic effect? AB - Congenital profound deafness has a known genetic origin in more than 50% of all cases. The majority of the non syndromic hearing loss (NSHL) show an autosomal recessive inheritance. Mutations in the GJB2 gene (connexin 26) account for more than 50% of the recessive non syndromic deafness (DFNB1) among 30 loci. Other connexin genes have been more rarely involved and attention was given here to the GJB6 gene (connexin 30). We show that homozygous deletion of a minimal 150 kb region encompassing this gene causes NSHL. More strikingly, association of this deletion in trans of the GJB2 gene 35delG or E47X mutations is also associated with NSHL. PMID- 11896457 TI - A detailed transcriptional map of the chromosome 12p12 tumour suppressor locus. AB - Loss of heterozygosity of the short arm of chromosome 12 is a frequent event in a wide range of haematological malignancies and solid tumours. In previous studies, the shortest commonly deleted region was delimited to a 750-kb interval, defined by the markers D12S89 and D12S358, in pre-B acute lymphoblastic leukaemia patients, suggesting the presence of a tumour suppressor locus. Here we report the construction of a transcriptional map that integrates the data obtained by genomic sequence analysis, EST database search, comparative analysis and exon amplification. We identified seven putative transcriptional units as well as six pseudogenes. Four of these candidate genes were already known: ETV6, encoding an ets-like transcription factor, LRP6, a member of the LDL receptor gene family, BCL-G, a recently identified pro-apoptotic gene and MKP-7, encoding a new member of the dual-specificity phosphatase family. The products encoded by the three new genes identified in this study, LOH1CR12, LOH2CR12 and LOH3CR12, have no clear homology to known proteins. The gene predictions were all confirmed by expression analysis using RT-PCR and Northern blot. This transcriptional map is a crucial step toward the identification of the tumour suppressor gene at 12p12. PMID- 11896459 TI - Somatic mosaicism for a MECP2 mutation associated with classic Rett syndrome in a boy. AB - Rett syndrome is a severe neurodevelopmental disorder that arises from mutations in the X-linked MECP2 gene. It is almost exclusively seen in girls due to the predominant occurrence of the mutations on the paternal X-chromosome, and also the early postnatal lethal effect of the disease causing mutations in hemizygous boys. We identified a boy with features of classic Rett syndrome who is mosaic for the truncating MECP2 mutation R270X. Chromosome analysis showed normal karyotype. These results indicate that a MECP2 mutation associated with Rett syndrome in females could lead to a similar phenotype in males as a result of somatic mosaicism. PMID- 11896460 TI - Relation between tumour necrosis factor polymorphism TNFalpha-308 and risk of asthma. AB - Tumour necrosis factor (TNF) alpha affects immune response and airway inflammation, which are characteristics of asthma. Genetic factors may impact TNFalpha levels, and several polymorphisms in the TNF gene cluster on chromosome 6p21 have been associated with TNFalpha production and potential increased risk of asthma. The present paper evaluates the relation between two single nucleotide polymorphisms (SNPs) in the TNF gene cluster and asthma risk. The SNPs investigated here are guanine (G) to adenosine (A) substitutions in the TNFalpha and lymphotoxin alpha (LTalpha) genes. The TNFalpha SNP is at position -308 in the promoter region (TNFalpha-308), while the LTalpha SNP is in the first intron NcoI recognition sequence (LTalpha-NcoI). (For both SNPs the G allele is denoted as 1, and the A allele 2.) We determined TNFalpha-308 and LTalpha-NcoI genotypes in 511 individuals: 236 asthma cases and 275 non-asthmatic controls. Data were analysed by logistic regression of asthma status on the genotypes and potential confounders. TNFalpha-308*2 was positively associated with asthma, and this relation was strengthened when restricting cases to individuals reporting acute asthma: the adjusted odds ratio (OR) comparing carriers of one or two TNFalpha 308*2 alleles versus none was 1.86 (95% confidence interval (CI)=1.03-3.34, P=0.04). Further restricting the subjects to those with a family history of asthma, and those of European-American ancestry strengthened the association even more: adjusted OR=3.16 (95% CI=1.04-9.66; P=0.04). LTalpha-NcoI*1 was weakly associated with asthma, and analysis of both genes suggests that only the TNFalpha-308*2 allele increases risk of asthma. PMID- 11896461 TI - Polymorphisms in the C-terminal domain of MECP2 in mentally handicapped boys: implications for genetic counselling. AB - Numerous recent reports have proposed that mutations in the C-terminal domain of the MECP2 gene could be a frequent cause of mental retardation in males. We have identified two mutations in this particular domain (S359P and E397K) in two boys who were screened for MECP2 mutations in a series of 23 mentally handicapped boys fitting the clinical description of the previously reported cases. A detailed familial study based on three generations shows that the first mutation (S359P) was also inherited by a healthy cousin thus ruling out its involvement in the etiology of the phenotype of this patient. The second mutation (E397K) was also found in normal individuals. These findings clearly call for a careful consideration of the pathogenicity of the MECP2 mutations identified in sporadic male cases before genetic counselling or prenatal diagnosis is proposed to the corresponding families. PMID- 11896462 TI - A novel conditional Akt 'survival switch' reversibly protects cells from apoptosis. AB - The anti-apoptotic Akt kinase is commonly activated by survival factors following plasma membrane relocalization attributable to the interaction of its pleckstrin homology (PH) domain with phosphatidylinositol 3-kinase (PI3K)-generated PI3,4 P(2) and PI3,4,5-P(3). Once activated, Akt can prevent or delay apoptosis by phosphorylation-dependent inhibition or activation of multiple signaling molecules involved in apoptosis, such as BAD, caspase-9, GSK3, and NF-kappaB and forkhead family transcription factors. Here, we describe and characterize a novel, conditional Akt controlled by chemically induced dimerization (CID). In this approach, the Akt PH domain has been replaced with the rapamycin (and FK506) binding domain, FKBP12, to make F3-DeltaPH.Akt. To effect membrane recruitment, a myristoylated rapamycin-binding domain from FRAP/mTOR, called M-FRB, binds to lipid permeable rapamycin (and non-bioactive synthetic 'rapalogs'), leading to reversible heterodimerization of M-FRB with FKBP-DeltaPH.Akt. Like endogenous c Akt, we show that the kinase activity of membrane-localized F3-DeltaPH.Akt correlates strongly with phosphorylation at T308 and S473; however, unlike c-Akt, phosphorylation and activation of inducible Akt (iAkt) is largely PI3K independent. CID-mediated activation of iAkt results in phosphorylation of GSK3, and contributes to NF-kappaB activation in vivo in a dose-sensitive manner. Finally, in Jurkat T cells stably expressing iAkt, CID-induced Akt activation rescued cells from apoptosis triggered by multiple apoptotic stimuli, including staurosporine, anti-Fas antibodies, PI3K inhibitors and the DNA damaging agent, etoposide. This novel inducible Akt should be useful for identifying new Akt substrates and for reversibly protecting tissue from apoptosis due to ischemic injury or immunological attack. PMID- 11896463 TI - Transduction of human neural progenitor cells using recombinant adeno-associated viral vectors. AB - Human neural progenitor cells (hNPCs) represent an attractive source for cell therapy of neurological disorders. Genetic modification of hNPCs may allow a controlled release of therapeutic proteins, suppress immune rejection, or produce essential neurotransmitters. In search of an effective gene delivery vehicle, we evaluated the efficiency of a recombinant adeno-associated viral (rAAV) vector expressing enhanced green fluorescent protein (CAGegfp). Our study demonstrated that CAGegfp efficiently transduced both proliferating and differentiated hNPCs in vitro. EGFP expression was detected as early as 1 day after exposure to CAGegfp and was detectable for up to 4 months. Following transduction, the growth rate of hNPCs slowed down, but they were still able to differentiate into neurons and glia. Furthermore, CAGegfp-modified hNPCs survived, differentiated and expressed EGFP after transplanting into spinal cord of adult rats. Our results indicated that rAAV vectors might be a useful tool in hNPC-based cell and gene therapy for neurological disorders. PMID- 11896464 TI - Effective single chain antibody (scFv) concentrations in vivo via adenoviral vector mediated expression of secretory scFv. AB - Single chain antibodies (scFv) represent powerful interventional agents for the achievement of targeted therapeutics. The practical utility of these agents have been limited, however, by difficulties related to production of recombinant scFv and the achievement of effective and sustained levels of scFv in situ. To circumvent these limitations, we have developed an approach to express scFv in vivo. An anti-erbB2 scFv was engineered for secretion by eukaryotic cells. The secreted scFv could bind to its target and specifically suppress cell growth of erbB2-positive cells in vitro. Adenoviral vectors expressing the cDNA for the secretory scFv likewise could induce target cells to produce an anti-tumor anti erbB2 scFv. In vivo gene transfer via the anti-erbB2 scFv encoding adenovirus also showed anti-tumor effects. Thus, by virtue of engineering a secreted version of the anti-tumor anti-erbB-2 scFv, and in vivo expression via adenoviral vector, effective concentrations of scFv were achieved. In vivo gene transfer clearly represents a powerful means to realize effective scFv-based approaches. This method will likely have applicability for a range of disorders amenable to targeted therapeutic approaches. PMID- 11896465 TI - Tumour cell radiosensitization using constitutive (CMV) and radiation inducible (WAF1) promoters to drive the iNOS gene: a novel suicide gene therapy. AB - Nitric oxide (NO(*)) has many characteristics including cytotoxicity, radiosensitization and anti-angiogenesis, which make it an attractive molecule for use in cancer therapy. We have investigated the use of iNOS gene transfer, driven by both a constitutive (CMV) and X-ray inducible (WAF1) promoter, for generating high concentrations of NO(*) within tumour cells. We have combined this treatment with radiation to exploit the radiosensitizing properties of this molecule. Transfection of murine RIF-1 tumour cells in vitro with the iNOS constructs resulted in increased iNOS protein levels. Under hypoxic conditions cells were radiosensitized by delivery of both constructs so that these treatments effectively eliminated the radioresistance observed under hypoxic conditions. In vivo transfer of the CMV/iNOS construct by direct tumour injection resulted in a delay (4.2 days) in tumour growth compared with untreated controls. This was equivalent to the effect of 20 Gy X-rays alone. Combination of CMV/iNOS gene transfer with 20 Gy X-rays resulted in a dramatic 19.8 day growth delay compared with controls. Tumours treated with the CMV/iNOS showed large areas of necrosis and abundant apoptosis. We believe that iNOS gene transfer has the potential to be a highly effective treatment in combination with radiotherapy. PMID- 11896466 TI - Adenoviruses with Tcf binding sites in multiple early promoters show enhanced selectivity for tumour cells with constitutive activation of the wnt signalling pathway. AB - Mutation of the adenomatous polyposis coli and beta-catenin genes in colon cancer leads to constitutive activation of transcription from promoters containing binding sites for Tcf/LEF transcription factors. We have constructed adenoviruses with Tcf binding sites in the early promoters, in order to target viral replication to colon tumours. Tcf regulation of the E1A promoter confers a 100 fold selectivity for cells with activated wnt signalling in viral burst and cytopathic effect assays. p300 is a coactivator for beta-catenin, and E1A inhibits Tcf-dependent transcription through sequestration of p300, but mutation of the p300 binding site in E1A leads to a 10-fold reduction in cytopathic effect of all of the Tcf-regulated viruses. When Tcf sites are inserted in the E1A, E1B, E2 and E4 promoters the viruses show up to 100 000-fold selectivity for cells with activated wnt signalling. PMID- 11896467 TI - Antibody-mediated lung endothelium targeting: in vivo model on primates. AB - We have recently provided evidence that angiotensin-converting enzyme (ACE) is a rational target and anti-ACE monoclonal antibodies (mAbs) are suitable molecules for directing gene/drug delivery into the pulmonary endothelium of rodents. As a step towards gene therapy clinical trials using this approach, the present study evaluated the potential of anti-ACE mAbs for in vivo lung endothelium targeting in 10 species of primates. Cross-reactivity of 10 distinct mAbs directed to human ACE with ACE from baboon, macaques, cercopithecus and chimpanzee revealed that the highest binding with ACE from baboon and macaques was with mAb i2H5, from chimpanzee - mAb 9B9, and from human - 9B9 and i2H5. Thereafter, in vivo biodistribution of mAbs i2H5 and 9B9 was estimated in Macaca arctoides. MAb i2H5, which binds to macaque ACE with substantially higher affinity than mAb 9B9, also more effectively accumulates in their lungs than mAb 9B9. Immunospecificity of lung accumulation (mAb/control IgG ratio) was 37 for i2H5 and 0.5 for 9B9. Lung selectivity of i2H5 uptake (lung/blood ratio) was around 10. Therefore mAb i2H5 may be useful for in vivo lung targeting in non-human primates, whereas 9B9 may be most useful in primates that are closer to humans (chimpanzee). A combination of these two mAbs may be particularly useful for human clinical trials of gene/drug therapy for lung disorders such as pulmonary hypertension and lung metastases. PMID- 11896468 TI - Anticancer efficacy of systemically delivered anaerobic bacteria as gene therapy vectors targeting tumor hypoxia/necrosis. AB - A major obstacle in cancer gene therapy is selective tumor delivery. Previous studies have suggested that genetically engineered anaerobes of the genus Clostridium might be gene therapy vectors because of their ability to proliferate selectively in the hypoxic/necrotic regions common to solid tumors. However, the tumor colonization efficiency of the strain previously used was insufficient to produce any antitumor effect. Here we describe for the first time the successful transformation of C. sporogenes, a clostridial strain with the highest reported tumor colonization efficiency, with the E. coli cytosine deaminase (CD) gene and show that systemically injected spores of these bacteria express CD only in the tumor. This enzyme can convert the nontoxic prodrug 5-fluorocytosine (5-FC) to the anticancer drug 5-fluorouracil (5-FU). Furthermore, systemic delivery of 5-FC into mice previously injected with CD-transformed spores of C. sporogenes produced greater antitumor effect than maximally tolerated doses of 5-FU. Since most human solid tumors have hypoxic and necrotic areas this vector system has considerable promise for tumor-selective gene therapy. PMID- 11896469 TI - A tricistronic retroviral vector expressing natural antiangiogenic factors inhibits angiogenesis in vitro, but is not able to block tumor progression in vivo. AB - Angiogenesis, the formation of new blood vessels out of pre-existing capillaries, is essential for tumor progression. Many factors have been identified that are able to inhibit angiogenesis. Here, we report the construction of a tricistronic retroviral vector encoding two inhibitors of angiogenesis expressed in mammals: the N-terminal fragment of rat prolactin (16KrPRL) and a secreted form of human platelet factor 4 (sPF4). When transduced by this retroviral vector, a rat glioblastoma cell line loses its ability of promoting endothelial cell locomotion, the initial step of angiogenesis, and the formation of an endothelial cell tube network. In spite of this encouraging in vitro result, however, the anti-angiogenic vector cannot block glioblastoma progression in animal models. These results suggest that therapeutic strategies aiming to block tumor progression through the inhibition of tumor-associated angiogenesis, should not only provide large numbers of angiogenesis inhibitors, but also target the angiogenic factors produced by tumor cells. Moreover, the data described herein may confirm recent findings from other groups which indicate that in order to successfully counteract tumor progression, drugs inhibiting new blood vessel formation should be employed in combination with traditional anti-tumor strategies, such as chemotherapy or radiotherapy. PMID- 11896470 TI - High expression of transgenes mediated by hybrid retroviral vectors in hepatocytes: comparison of promoters from murine retroviruses in vitro and in vivo. AB - To achieve high transgene expression in the liver, we have compared the reporter gene expression among various murine retroviral long terminal repeats (LTRs) or leader sequences in vitro. Transient reporter gene expression assays revealed the highest gene expression by the polycythemic strain of spleen focus-forming virus (SFFVp) LTR in differentiated hepatocellular carcinoma cell lines, HuH-7 and PLC/PRF/5. However, remarkable difference was not observed among LTRs in other types of human liver tumor cell lines. Essentially the same results were obtained by infecting these cells with a series of retroviral vectors. Repression of transgene expression was observed by the leader sequences from Moloney murine leukemia virus (MoMLV), but not from mouse embryonic stem cell virus (MESV). Strengths of the promoters were further compared in murine hepatocytes in vivo. Although the proportions of genomic integration were almost the same, higher gene expression was observed by the FMEV-type vector, which contained the SFFVp LTR and the MESV leader, in comparison with that by the MoMLV-based vector. Thus, FMEV-type vectors may represent a novel type of vectors for human gene therapy with hepatocytes. PMID- 11896471 TI - Human and rabbit cavernosal smooth muscle cells express Rho-kinase. AB - Rho-kinase is an enzyme involved in the Ca2+-sensitizing pathway in smooth muscle cells. Inhibition of this enzyme has been recently demonstrated to elicit penile erection by relaxing cavernosal smooth muscle. We aimed to investigate the presence and activity of Rho-kinase in human cavernosal smooth muscle. Primary culture of smooth muscle cells from human and rabbit penile corpus cavernosum was developed, and cells showed characteristic myocyte morphology and alpha-actin immunoreactivity. The presence of Rho-kinase was demonstrated by indirect immunofluorescence and Western blotting. A specific inhibitor of Rho-kinase, Y 27632, inhibited in a concentration-dependent manner the kinase activity of the protein immunoprecipitated with anti-Rho-kinase antibody. These results demonstrate for the first time expression and activity of Rho-kinase in human penile cavernosal smooth muscle cells and suggest that these cells can provide a cellular model for the study of enzymes involved in Ca2+-sensitizing pathways. PMID- 11896472 TI - Activators of soluble guanylate cyclase for the treatment of male erectile dysfunction. AB - Soluble guanylate cyclase (sGC) is an important enzyme in corpus cavernosum smooth muscle cells as it is one of the regulators of the synthesis of cGMP. The efficacy of sildenafil (Viagra) in the treatment of male erectile dysfunction indicates the importance of the cGMP system in the erectile response as the increased levels of cGMP induce relaxation of the corpus cavernosum. sGC is physiologically activated by nitric oxide (NO) during sexual stimulation, and its activity can be pharmacologically enhanced by several NO-donors. Agents like YC-1 can also activate sGC after binding to a novel allosteric site in the enzyme, a site different from the NO binding site. YC-1 can relax rabbit cavernosal tissue and it facilitates penile erection in vivo. This review summarizes the enzymology, biochemistry and pharmacology of this novel allosteric site and its relevance for the regulation of penile function. This type of sGC activators represent a new class of compounds with a different pharmacological profile in comparison to the classical NO-donors and they could be beneficial for the treatment of male erectile dysfunction. PMID- 11896473 TI - Human PDE5A gene encodes three PDE5 isoforms from two alternate promoters. AB - Sildenafil improves erectile function by inhibiting the cGMP-catalytic activity of phosphodiesterase type V (PDE5). We used rapid amplification of cDNA Ends polymerase chain reaction (RACE-PCR) to isolate three PDE5 isoforms from human corpus cavernosum. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis on eight human cavernous tissue samples showed that all samples expressed the PDE5A1 at a lower level than the PDE5A2 isoform. Five samples expressed the PDE5A3 isoform at various levels while the other three did not. Analysis on non-penile tissues showed that all tissues expressed the A1 and A2 isoforms while only those that have substantial amounts of smooth muscle expressed the A3 isoform. Cloning and sequencing of the PDE5A gene showed that the isoform-specific 5'-ends of the PDE5 mRNAs are encoded from three alternative first exons arranged in the order of A1-A3-A2. Promoter activities were detected upstream from the A1-specific exon and in the intron preceding the A2-specific exon. The upstream PDE5A promoter is expected to direct the expression of all three PDE5 isoforms while the intronic PDE5A2 promoter only the A2 isoform. Both promoters were upregulated by increasing concentrations of either cAMP or cGMP. Several transcription factor AP2 and Sp1-binding sequences identified in the promoters are likely to be the mediators of cAMP/cGMP-responsiveness. PMID- 11896474 TI - Yohimbine treatment of organic erectile dysfunction in a dose-escalation trial. AB - Yohimbine has had questionable effects in men with organic erectile dysfunction. We conducted this study to better define the population of men responsive to yohimbine, because tobacco was thought to affect a regimen of yohimbine more than other risk factors. We measured nocturnal penile tumescence with the RigiScan monitor, hormone profiles, answers to the Florida Sexual Health Questionnaire, and clinical responses at baseline and after two different doses of yohimbine in 18 nonsmoking men with erectile dysfunction. Of the 18 men, nine (50%) were successful in completing intercourse in more than 75% of attempts. The yohimbine responders were men with less severe erectile dysfunction as manifested by improved increased rigidity on RigiScan testing, higher Florida Sexual Health Questionnaire scores, and slightly higher levels of serum testosterone. Yohimbine is an effective therapy to treat organic erectile dysfunction in some men with erectile dysfunction. PMID- 11896475 TI - Role of penile vascular insufficiency in erectile dysfunction in renal transplant recipients. AB - The objectives of this study were to define the role and haemodynamic features of penile vascular insufficiency in impotent renal transplant recipients (RTR) as well as to establish the possible vascular risk factors for impotence in these patients. A total of 54 RTR (35 impotent and 19 potent) and 21 potent healthy subjects were included in this study. All patients were assessed clinically and by measurement of serum creatinine, serum bilirubin, cyclosporine blood levels, haemoglobin and total serum cholesterol. All subjects were subjected to intracavernous injection of 20 microg prostaglandin E1 followed by colour Duplex sonographic examination. Our results showed that impotent RTR were significantly more likely than potent RTR to have hypertension, diabetes and hypercholesterolaemia (P<0.05). Arterial occlusive disease was identified in 42.9% of impotent RTR. Findings suggestive of veno-occlusive dysfunction were found in 68.6% and 26.3% of impotent and potent RTR, respectively (P=0.003). Unilateral ligation of the internal iliac artery has a negative role on haemodynamic parameters compared to unilateral end-to-side anastomosis to external iliac artery in impotent RTR (P<0.05). Impotent RTR receiving more than one antihypertensive drug showed significant decrease in basal peak systolic velocity (PSV), dynamic PSV, erectile angle and cavernosal artery diameter compared to those receiving one drug (P<0.05). In conclusion, penile vascular insufficiency appears to play a substantial role in the pathogenesis of impotence in transplant patients. Anastomosis of the graft to external iliac artery could preserve the potency to some degree. Antihypertensives should be reduced as much as possible to avoid their negative effects on erectile function. PMID- 11896476 TI - Intracavernous injections for erectile dysfunction in patients with cardiovascular diseases and failure or contraindications for sildenafil citrate. AB - The aim of this study was to evaluate the effectiveness of a progressive program for the treatment of erectile dysfunction in patients with cardiovascular disease in whom sildenafil citrate (Viagra) was not an option. The study population included 106 patients selected from 267 with cardiovascular disease. The intracavernous injection program consisted of three protocols of increasingly complex combinations of vasoactive drugs, papaverine, phentolamine, prostaglandin E1 and atropine sulfate. Patients who failed the first protocol were switched to the second, and those who failed the second were switched to the third. A positive response was defined as an erection sufficient for vaginal penetration. A positive response was achieved on protocol I in 61 of the 106 patients (57.5%); protocol II in 32 of the remaining 45 patients (71.1%); and protocol III in seven of the remaining 13 patients (53.8%); the total success rate was 94.3%. These 100 patients were included in the 1-year follow-up, and 90 reported successful coitus at the end of that period: 79 patients (87.8%) with intracavernous injection and 11 (12.2%) without injection. The remaining 10 patients (10%) dropped out of the program, seven (7.0%) for health or marital reasons and three (3.0%) because of treatment failure. We conclude that a progressive program of intracavernous injections of vasoactive drugs may be a good alternative for the treatment of erectile dysfunction in patients with cardiovascular disease. PMID- 11896477 TI - Intracavernous administration of SIN-1+VIP in an in vivo rabbit model for erectile function. AB - In accordance with the data reporting the identification of nitric oxide synthase (NOS) and vasoactive intestinal polypeptide (VIP) positive nerve fibres in the trabecular meshwork of the corpus cavernosum, we suggest that nitric oxide (NO) and VIP may serve complementary physiological roles in penile erection. Therefore SIN-1 and VIP were administered alone and in combination in an in vivo rabbit model. All rabbits revealed basal pressure values of 5-8 cm H2O intracavernously. In the rabbits intracavernously (i.c.) injected with SIN-1 alone and VIP alone, no adequate erectile responses were observed. Whereas, in the group intracavernously injected with the combination of SIN-1+VIP, erectile responses with mean maximal intracavernous pressure (max. ICP) 52.8 (+/-13.2) cm H2O were noted. These pressure elevations do not statistically diverge (P>0.05) than the ones obtained in the control group administered i.c. injections of the combination of papaverine/phentolamine (mean max. ICP 51 (+/-14.73) cm H2O). Referring to our results, we conclude that the combined use of SIN-1+VIP could play an important role in the physiological treatment of erectile dysfunction. PMID- 11896478 TI - Combined oral therapy with sildenafil and doxazosin for the treatment of non organic erectile dysfunction refractory to sildenafil monotherapy. AB - The purpose of this work was to investigate the efficacy and safety of sildenafil in combination with doxazosin for the treatment of non-organic erectile dysfunction in patients who did not respond to sildenafil. We enrolled 28 patients with non-organic erectile dysfunction, for whom 3 months of sildenafil monotherapy had failed. They were divided in two random and homogeneous groups: 14 were treated with doxazosin (4 mg daily) and sildenafil (100 mg 1 h before sexual intercourse); the other 14 patients received sildenafil and placebo. The results were assessed by means of the IIEF questionnaire before the beginning of the study, after 30 days of therapy and after 60 days. Of the 14 patients treated with doxazosin and sildenafil, 11 (78.6%) showed a statistically significant increase of IIEF; in the placebo group, only one patient (7.1%) recorded a significant IIEF increase. The differences observed in the two groups were statistically very significant (P=0.0016). Blood pressure did not show significant alterations. Side effects were minimal and even present during sildenafil monotherapy. The combination therapy with sildenafil and doxazosin resulted in the safe and effective treatment of men with non-organic erectile dysfunction for whom sildenafil alone had failed. PMID- 11896479 TI - Combination therapy for erectile dysfunction: a randomized, double blind, unblinded active-controlled, cross-over study of the pharmacodynamics and safety of combined oral formulations of apomorphine hydrochloride, phentolamine mesylate and papaverine hydrochloride in men with moderate to severe erectile dysfunction. AB - Oral therapy has become first line treatment for patients with mild to moderate erectile dysfunction (ED). Studies have shown that sildenafil may not be effective in all patients, and has been associated with a variety of adverse effects and an adverse interaction with nitrates and inhibitors of cytochrome P450 enzymes. The objective was to compare the efficacy and safety of three different oral combinations with the highest dose of sildenafil in men with moderate to severe ED. Randomized, double blind, unblinded active-controlled, Phase II study was carried out at three sites in Mexico. After a 4-week placebo run-in period, patients received all four of the following treatments using a 4 way cross-over design: 40 mg phentolamine (PM) +6 mg apomorphine (Apo); 40 mg PM +150 mg papaverine (Pap); 40 mg PM +6 mg Apo +150 mg Pap (Tricombo); 100 mg sildenafil (SC). With the exception of sildenafil tablets, all study medication was blinded. Moderate to severe ED was defined as a less than 50% vaginal penetration success rate during the placebo run-in period. A total of 44 patients were enrolled, of whom 36 completed all four treatment periods. All treatments produced a significant effect in primary efficacy variable (Sexual Encounter Profile) compared to baseline, however, no statistically significant differences were found between treatments. A significant period effect was observed. Also, the four treatments were found not to differ significantly in five out of six secondary efficacy variables. The lowest incidence of treatment-related adverse events (AE) occurred in the 40 mg PM +6 mg Apo group (9.8%), followed by 100 mg SC (15%), and the other two combinations (16.7 and 17.5%, respectively). Nasocongestion and headache were the most frequently reported AE. An oral combination of vasoactive agents may provide an alternative approach to sildenafil. Based on these results a combination of phentolamine and apomorphine warrants further clinical investigation. PMID- 11896482 TI - Sequential administration enhances the effect of apomorphine SL in men with erectile dysfunction. AB - The response to Uprima (apomorphine sublingual, (apo SL)) has been well documented in conventional clinical trials. Apo SL produces a predictable, consistent and durable response across a wide variety of patients. The positive reinforcement of a successful outcome should further support clinical benefit. Apo SL with its rapid onset affords a greater opportunity for spontaneity, which can be an important factor in influencing patient choice. It is recognised that patient counselling and the setting of realistic expectations are vital to a successful outcome. The impact of persisting with sequential treatment on outcome has been calculated from the clinical data. While apo SL is effective de novo in 50% of single doses, additional benefit is observed with repeat dosing. Full benefit may not be achieved until four or more treatments have been taken in an optimal setting. The data also confirm that 3 mg has superior activity. Patients should therefore be encouraged to try a minimum of 4 doses at 3 mg. PMID- 11896483 TI - Appetite suppression based on selective inhibition of NPY receptors. AB - AIM: The aim of this review is to critically assess available evidence that blockade of the actions of NPY at one of the five NPY receptor subtypes represents an attractive new drug discovery target for the development of an appetite suppressant drug. RESULTS: Blockade of the central actions of NPY using anti-NPY antibodies, antisense oligodeoxynucleotides against NPY and NPY receptor antagonists results in a decrease in food intake in energy-deprived animals. These results appear to show that endogenous NPY plays a role in the control of appetite. The fact that NPY receptors exist as at least five different subtypes raises the possibility that the actions of endogenous NPY on food intake can be adequately dissociated from other effects of the peptide. Current drug discovery has produced a number of highly selective NPY receptor antagonists which have been used to establish the NPY Y(1) receptor subtype as the most critical in regulating short-term food intake. However, additional studies are now needed to more clearly define the relative contribution of NPY acting through the NPY Y2 and NPY Y5 receptors in the complex sequence of physiological and behavioral events that underlie the long-term control of appetite. CONCLUSIONS: Blockade of the NPY receptor may produce appetite-suppressing drugs. However, it is too early to state with certainty whether a single subtype selective drug used alone or a combination of NPY receptor selective antagonists used in combination will be necessary to adequately influence appetite regulation. PMID- 11896484 TI - Binge eating disorder: a review. AB - Binge eating disorder (BED) is a new proposed eating disorder in the DSM-IV. BED is not a formal diagnosis within the DSM-IV, but in day-to-day clinical practice the diagnosis seems to be generally accepted. People with the BED-syndrome have binge eating episodes as do subjects with bulimia nervosa, but unlike the latter they do not engage in compensatory behaviours. Although the diagnosis BED was created with the obese in mind, obesity is not a criterion. This paper gives an overview of its epidemiology, characteristics, aetiology, criteria, course and treatment. BED seems to be highly prevalent among subjects seeking weight loss treatment (1.3-30.1%). Studies with compared BED, BN and obesity indicated that individuals with BED exhibit levels of psychopathology that fall somewhere between the high levels reported by individuals with BN and the low levels reported by obese individuals. Characteristics of BED seemed to bear a closer resemblance to those of BN than of those of obesity.A review of RCT's showed that presently cognitive behavioural treatment is the treatment of choice but interpersonal psychotherapy, self-help and SSRI's seem effective. The first aim of treatment should be the cessation of binge eating. Treatment of weight loss may be offered to those who are able to abstain from binge eating. PMID- 11896485 TI - Developmental changes in adipose and muscle lipoprotein lipase activity in the atherosclerosis-prone JCR:LA-corpulent rat. AB - OBJECTIVE: To characterize the developmental changes in adipose and muscle lipoprotein lipase (LPL) activity in the atherosclerosis-prone JCR:LA-corpulent rat, and to test the hypothesis that tissue-specific abnormalities in LPL activity precede the establishment of obesity. DESIGN: Lean (+/?) and obese cp/cp male JCR:LA rats were studied at 4, 5 and 8 weeks of age, that is at the onset of obesity, and at a time when obesity is well established. Assessment was made of plasma variables related to glucose and lipid metabolism and of LPL activity in several adipose depots, skeletal muscles and the heart. RESULTS: At week 4, body weights were identical in both genotypes and began to diverge at week 5. Eight week-old cp/cp rats weighed 35% more than their lean counterparts. Perirenal and epididymal adipose depot weights were also identical in both genotypes at week 4 and began to increase in cp/cp rats at week 5, whereas the subcutaneous depot of 4-week-old cp/cp rats was slightly enlarged. At week 4, the cp/cp rats were hyperinsulinemic (5-fold), hyperleptinemic (30-fold) and hypertriglyceridemic (3 fold) compared to their lean counterparts, and their liver contained twice as much triglyceride. The 4-week-old cp/cp rats displayed 2-7-fold higher LPL specific activity in the various adipose depots compared to lean rats, and enzyme activity remained higher in obese than in lean rats at all subsequent ages. In contrast, LPL activity in the vastus lateralis, gastrocnemius and heart muscles of 4-week-old obese rats was approximately half that observed in lean animals. CONCLUSION: Profound, persistent alterations in the tissue-specific modulation of LPL activity are established in the JCR:LA cp/cp rat prior to the development of frank obesity. The increase in adipose tissue LPL activity and its decrease in muscle tissues are likely to be related to the concomitant alterations in insulinemia and triglyceridemia, respectively. The pre-obesity, tissue-specific alterations in LPL activity may be considered as an integrated adaptation to increased lipid flux aimed at driving lipids toward storage sites and limiting their uptake by triglyceride-laden muscles. PMID- 11896486 TI - Early events involved in the development of insulin resistance in Zucker fatty rat. AB - AIM: To clarify the mechanism by which insulin resistance develops in obesity, Zucker fatty rats (ZFR) and lean litter mates (ZLR) were temporally subjected to oral glucose tolerance tests (OGTT) at 6 and 15 weeks of age. METHOD: As candidates for causative factors of insulin resistance, plasma leptin, free fatty acids (FFA) and tumor necrosis factor (TNF)-alpha levels were evaluated. RESULTS: There was no difference in the body weight between the two groups at 6 weeks of age, but ZFR were significantly heavier than ZLR at 15 weeks of age. At 6 weeks of age, blood glucose levels and area under the curve of glucose (AUCg) during OGTT were not significantly different between the two groups, while plasma insulin levels and area under the curve of insulin (AUCi) in the ZFR group were significantly higher than those in the ZLR group. At 15 weeks of age, the blood glucose levels and AUCg as well as plasma insulin levels and AUCi in the ZFR group during OGTT were significantly higher than those in the ZLR group. The ratio of fasting insulin to glucose in the ZFR group was significantly higher than that in the ZLR group at 6 and 15 weeks of age. Peripheral and portal plasma leptin and FFA levels were significantly higher in ZFR than ZLR both at 6 weeks and 15 weeks of age. Meanwhile, at 6 weeks, plasma TNF-alpha levels and expression of TNF-alpha protein in subcutaneous and visceral fat tissues were similar in both groups; however at 15 weeks, these were significantly higher in the ZFR group than the ZLR group. CONCLUSION: These results suggest that FFA rather than TNF-alpha may play an important role in early events involved in the development of insulin resistance and TNF-alpha accelerates insulin resistance together with FFA in the later stage. PMID- 11896487 TI - Impaired glucose metabolism in the heart of obese Zucker rats after treatment with phorbol ester. AB - OBJECTIVE: To investigate the influence of obesity on the regulation of myocardial glucose metabolism following protein kinase C (PKC) activation in obese (fa/fa) and lean (Fa/?) Zucker rats. DESIGN: Isolated hearts obtained from 17-week-old lean and obese Zucker rats were perfused with 200 nM phorbol 12 myristate 13-acetate (PMA) for different time periods prior to the evaluation of PKC and GLUT-4 translocation. For metabolic studies isolated hearts from 48 h starved Zucker rats were perfused with an erythrocytes-enriched buffer containing increased concentrations (10-100 nM) of PMA. MEASUREMENTS: Immunodetectable PKC isozymes and GLUT-4 were determined by Western blots. Glucose oxidation and glycolysis were evaluated by measuring the myocardial release of 14CO2 and 3H2O from [U-14C]glucose and [5-3H]glucose, respectively. RESULTS: PMA (200 nM) induced maximal translocation of ventricular PKCalpha from the cytosol to the membranes within 10 min. This translocation was 2-fold lower in the heart from obese rats when compared to lean rats. PMA also induced a significant translocation of ventricular GLUT-4 from the microsomal to the sarcolemmal fraction within 60 min in lean but not in obese rats. Rates of basal cardiac glucose oxidation and glycolysis in obese rats were approximately 2-fold lower than those of lean rats. Perfusion with increasing concentrations of PMA (10-100 nM) led to a significant decrease of cardiac glucose oxidation in lean but not in obese rats. CONCLUSION: Our results show that in the heart of the genetically obese Zucker rat, the impairment in PKCalpha activation is in line with a diminished activation of GLUT-4 as well as with the lack of PMA effect on glucose oxidation. PMID- 11896488 TI - Effects of physical training on body composition and organ weights in ovariectomized and hyperestrogenic rats. AB - OBJECTIVE: To investigate whether regular endurance-type exercise can benefit rats submitted to a model of ovariectomy (OVX)-induced obesity with or without estrogen replacement. SUBJECTS: OVX Sprague-Dawley rats were compared to an ovariectomized-estradiol-treated group (OVXE2) and a Sham-operated (Sham) group. Each of these groups were subdivided into a sedentary and a treadmill-trained (8 wk) group. DESIGN AND MEASUREMENTS: An experimental study in which various parameters, including fat depots, blood lipids and several organ weights were measured. RESULTS: Plasma levels of 17beta-estradiol and uterus weights were significantly (P<0.05) lower in OVX compared to Sham and significantly (P<0.01) higher in OVXE2 (hyperestrogenic) compared to Sham rats. Body weights were significantly (P<0.01) different among groups, in the following decreasing order: OVX, Sham and OVXE2. The average daily food intake and food efficiency were significantly (P<0.01) increased in OVX compared to Sham, whereas estradiol treatment diminished this effect (P<0.01). Exercise training did not alter any of the above-mentioned variables in any of the three estrogen groups. Mesenteric and subcutaneous fat weights were significantly (P<0.01) increased by OVX. This increase was abolished by estrogen replacement or by exercise training. Exercise training also decreased fat weights in OVXE2 and Sham rats. OVX resulted in a decrease in the weights of several other tissues (femur, heart, lungs, liver and adrenal glands) while hyperestrogenic replacement resulted in an increase in weight of all measured tissues. Aside from fat depots, exercise training did not affect any of the tissue weights with the exception for an increase in the weight of the plantaris muscle and adrenal glands and a decrease in lung weight in all three estrogen groups. CONCLUSION: In OVX animals, exercise training may bring about positive changes in body composition (ie reduction in fat weights) despite an ovariectomy-induced increase in body weight. PMID- 11896489 TI - The effects of topiramate and sex hormones on energy balance of male and female rats. AB - OBJECTIVE: The effects of topiramate (TPM) on components of energy balance were tested in male and female rats that were (i) left intact, (ii) castrated or (iii) castrated with replacement therapies consisting of testosterone administration in orchidectomized (OCX) rats and of estradiol or progesterone treatments in ovariectomized (OVX) rats. METHODS: TPM was mixed into the diet and administered at a dose of 60 mg per kg of body weight. Male and female rats were treated for 28 and 35 days, respectively. At the end of the treatment period, variables of energy balance and determinants of lipid and glucose metabolism were assessed. RESULTS: TPM reduced energy and fat gains in both male and female rats either in the absence or in the presence of hormone replacement therapies. In male rats, it also decreased food intake, protein gain and energetic efficiency. In female animals, TPM reduced energetic efficiency while it stimulated lipoprotein lipase activity in brown adipose tissue. TPM also reduced plasma glucose and plasma leptin levels in female rats as well as plasma insulin and liver triglycerides in male animals. As expected, castration and sex hormones also strongly influenced energy balance. In male rats, OCX led to a decrease in energy and protein gains that was blocked by treatment with testosterone. In female rats, OVX caused increases in energy, fat and protein gains that were prevented by treatment with estradiol. CONCLUSION: In female rats, the effects of TPM on fat and energy gains were clearly not influenced by the sex hormone status of the rats. In male animals, there was also no interaction of TPM and the status of sex hormones on energy balance, suggesting that OCX and testosterone minimally interfere with the action of TPM on energy balance. The effects of TPM on energy balance were accounted for by a decrease in energetic efficiency, resulting from an effect exerted by the drug on both energy intake and thermogenesis. The present results also suggest that TPM can enhance insulin sensitivity. PMID- 11896490 TI - Sibutramine-dependent brown fat activation in rats: an immunohistochemical study. AB - OBJECTIVES: To evaluate morphological aspects and immunohistochemical markers of brown adipose tissue (BAT) activation following chronic treatment with sibutramine, a novel anti-obesity drug which increases thermogenesis and energy expenditure in mammals, and to establish whether chronic sibutramine treatment induces recruitment of BAT in white adipose tissue (WAT) depots. DESIGN: Adult rats were administered 7 mg/kg/day oral sibutramine for 4 weeks. Body weight was monitored daily. At the end of the 4 weeks rats were perfused with buffered paraformaldehyde solution; interscapular BAT and retroperitoneal and epididymal WAT were carefully dissected for weight and volume measurements and processed for light microscopic studies and immunohistochemistry on paraffin-embedded sections. Where possible, semiquantitative morphometric analyses were performed. RESULTS: Chronic sibutramine treatment determined a significant (about 8%) reduction in body weight. Compared with controls, sibutramine-treated rats showed: (1) interscapular brown adipocytes staining more intensely for uncoupling protein 1 (UCP1), the thermogenic mitochondrial protein; (2) a significantly larger number (about 45%) of brown adipocyte nuclei positive for peroxisome proliferator activated receptor gamma, the transcription factor driving UCP1 expression; (3) surprisingly, a significant reduction (about 30%) in BAT parenchymal noradrenergic nerve staining; and (4) a significant weight and volume reduction of WAT depots, but no significant signs of transdifferentiation of white into brown adipocytes. CONCLUSION: This study confirms the ability of sibutramine to induce weight loss by selective and sustained activation of BAT in rodents without recruitment of brown fat in WAT depots. The parallel findings of a high level of brown adipocyte activation and low parenchymal noradrenergic innervation are discussed and a possible direct effect of sibutramine and/or its active metabolites on peripheral BAT sympathetic nerve terminals is hypothesized. PMID- 11896491 TI - Relationship between reduced serum IGF-I levels and accumulation of visceral fat in Japanese men. AB - OBJECTIVE: To investigate whether the changes in IGF-I concentrations after weight reduction in Japanese overweight men are associated with changes in visceral and subcutaneous fat. DESIGN: Cross-sectional and longitudinal clinical intervention study with exercise education. SUBJECTS: One-hundred and twelve Japanese overweight men aged 30-59 y (body mass index (BMI) 28.4+/-2.5 kg/m(2)) and 33 normal-weight men aged 30-39 y (BMI 22.1+/-1.5 kg/m(2)) at baseline. From the participants, 56 randomly selected overweight men (BMI 28.8+/-2.8) were further enrolled into a 1 y exercise program. MEASUREMENTS: Fat distribution was evaluated by visceral fat (V) and subcutaneous fat (S) areas measured with computed tomography scanning at umbilical levels, metabolic parameters and hormones including insulin, leptin and IGF-I at baseline and after 1 y. RESULTS: In 112 overweight subjects at baseline, insulin (10.5+/-5.0 microU/ml) and leptin (6.4+/-3.7 ng/ml) significantly correlated with both V (r=0.260, P=0.0073; r=0.410, P<0.0001) and S areas (r=0.377, P<0.0001; r=0.613, P<0.0001), respectively. IGF-I (156.8+/-48.7 microU/ml) significantly and negatively correlated with V area (r=-0.242, P=0.0125) and age (r=-0.192, P=0.0480). In normal-weight men aged 30-39 y (n=33) and age-matched subjects (n=30) selected from the 112 overweight men, the serum IGF-I further tightly correlated with V area (r=-0.467, P<0.0001). Visceral fat area and age were independently related to serum IGF-I levels by multiple regression analysis. By intervention with exercise education, 56 overweight subjects showed an increase in daily steps (6224+/-2781 to 7898+/-4141 steps/day) and reduction of BMI (28.8+/-2.8 to 27.7+/ 2.9). deltaIGF-I significantly correlated with deltaV area (r=-0.432, P=0.0009) but not with DeltaS area or deltaBMI. CONCLUSION: The present study indicated a negative correlation between IGF-I levels and visceral fat at baseline as well as an association between the reduction in visceral fat and increase in IGF-I levels after an exercise intervention. PMID- 11896492 TI - Soluble leptin receptor and insulin resistance as determinant of sleep apnea. AB - OBJECTIVE: To investigate the possible associations between sleep apnea syndrome, hyperinsulinemia/insulin resistance and hyperleptinemia in subjects with different degrees of body mass index. DESIGN: To test for the presence or absence of sleep apnea syndrome in association with hyperinsulinemia/insulin resistance and hyperleptinemia. SUBJECTS: Twenty subjects with different body mass index (mean BMI 30.9+/-4.2). MEASUREMENTS: Insulin action and plasma soluble leptin receptor were measured by euglycemic hyperinsulinemic glucose clamp and by ELISA method, respectively. Occurrence of sleep apnea syndrome was assessed by clinical and nocturnal monitoring using a validated sleep apnea recorder. RESULTS: The apnea/hypopnea index (AHI) was positively correlated with plasma soluble leptin receptor (0.76; P<0.001) and negatively with the degree of insulin-mediated glucose uptake (r=-0.73; P<0.001). In a multivariate analysis AHI was associated with plasma soluble leptin receptor and insulin mediated glucose uptake independently of age, gender, BMI, plasma leptin levels and PaCO(2). CONCLUSION: Sleep apnea syndrome is associated with plasma soluble leptin receptor and insulin resistance independently of BMI. PMID- 11896493 TI - Sleeping metabolic rate in relation to body mass index and body composition. AB - OBJECTIVES: To determine whether patterns of sleeping metabolic rate (SMR) are altered in obesity. Specifically to determine the relationship between changes in SMR and body weight, body mass index (BMI, kg/m(2)), and fat-free mass (FFM); and to compare resting metabolic rate (RMR) with SMR during different periods of sleep. SUBJECTS: Eighteen healthy, pre-menopausal, obese (BMI >30, n=9) and non obese (BMI <30, n=9), female subjects (six Caucasians and 12 African-Americans), with an average age of 36 y (range 22-45). MEASUREMENTS: Total energy expenditure (TEE or 24 h EE), metabolic rate (MR), SMR (minimum, average and maximum) and resting metabolic rate (RMR) or resting energy expenditure (REE) measured by human respiratory chamber, and external mechanical work measured by a force platform within the respiratory chamber. Physical activity index (PAL) was derived as TEE/REE. Body composition was determined by dual-energy X-ray absorptiometry (DXA). RESULTS: SMR decreased continuously during sleep and reached its lowest point just before the subject was awakened in the morning by the research staff. Although averages for RMR and SMR were similar, RMR was lower than SMR at the beginning of the sleeping period and higher than SMR in the morning hours. The rate of decrease in SMR was faster with increasing body weight (-0.829, P<0.0001), BMI (correlation factor -0.896, P<0.0001) and FFM (-0.798, P=0.001). The relationship between the slope of SMR decrease and BMI (y=-5 x 10( 6)x(2)+0.0002x-0.0028) is highly significant, with a P-value of <0.0001 and r(2) value of 0.9622. CONCLUSIONS: The rate of decline in metabolic rate during sleep is directly related to body weight, BMI and FFM. Average SMR tends to be lower than RMR in obese subjects and higher than RMR in non-obese subjects. PMID- 11896494 TI - An investigation of the role of oro-sensory stimulation in sugar satiety? AB - OBJECTIVE: To investigate whether oral factors stimulated by the presence of sucrose in the mouth are involved in the suppression of appetite following sucrose ingestion. DESIGN: Ten male and 10 female healthy volunteers participated in four experimental conditions designed to provide differing levels of oro sensory stimulation. Appetite and energy intake from a test meal were measured after subjects chewed and ingested sucrose-containing pastilles over a 10 min period, consumed a sucrose-containing jelly over a 5 min period, consumed a sucrose-containing drink within 2 min and drank plain water within 2 min. The three sucrose-containing preloads were similar in nutrient composition, each containing 251 kJ. RESULTS: Ratings of hunger and fullness did not differ between the four conditions following ingestion of the preloads. However, energy intake from a test lunch was significantly reduced after consuming the pastilles when compared with the plain water and equicaloric sweet drink conditions. CONCLUSION: These results suggest that enhanced oro-sensory stimulation from chewing the sweet food was involved in the suppression of food intake. PMID- 11896495 TI - Comparison of percent body fat estimates using air displacement plethysmography and hydrodensitometry in adults and children. AB - PURPOSE: The purpose of the study was to compare estimates of body density and percentage body fat from air displacement plethysmography (ADP) to those from hydrodensitometry (HD) in adults and children and to provide a review of similar recent studies. METHODS: Body density and percentage body fat (% BF) were assessed by ADP and HD on the same day in 87 adults aged 18-69 y (41 males and 46 females) and 39 children aged 8-17 y (19 males and 20 females). Differences between measured and predicted thoracic gas volumes determined during the ADP procedure and the resultant effects of those differences on body composition estimates were also compared. In a subset of 50 individuals (31 adults and 19 children), reliability of ADP was measured and the relative ease or difficulty of ADP and HD were probed with a questionnaire. RESULTS: The coefficient of reliability between %BF on day 1 and day 2 was 96.4 in adults and 90.1 in children, and the technical error of measurement of 1.6% in adults and 1.8% in children. Using a predicted rather than a measured thoracic gas volume did not significantly affect percentage body fat estimates in adults, but resulted in overestimates of percentage body fat in children. Mean percentage body fat from ADP was higher than percentage body fat from HD, although this was statistically significant only in adults (29.3 vs 27.7%, P<0.05). The 95% confidence interval of the between-method differences for all subjects was -7 to +9% body fat, and the root mean square error (r.m.s.e.) was approximately 4% body fat. In the subset of individuals who were asked to compare the two methods, 46 out of 50 (92%) indicated that they preferred the ADP to HD. CONCLUSION: ADP is a reliable method of measuring body composition that subjects found preferable to underwater weighing. However, as shown here and in most other studies, there are differences in percentage body fat estimates assessed by the two methods, perhaps related to body size, age or other factors, that are sufficient to preclude ADP from being used interchangeably with underwater weighing on an individual basis. PMID- 11896496 TI - A novel method of measuring intra-abdominal fat volume using helical computed tomography. AB - OBJECTIVES: :We present a novel method of scanning for intra-abdominal fat volume by helical computed tomography (CT), and describe the clinical significance of measuring the volumes of intra-abdominal visceral fat (V(vol)) and subcutaneous fat (S(vol)) vs these respective areas determined by conventional slice-by-slice CT at the umbilical level. METHOD: Subjects with obesity or hyperlipidemia (79 men, 74 women) were recruited for this study. We obtained helical CT scans with a tube current of 150 mA, voltage of 120 kV and 2:1 pitch (table speed in relation to slice thickness), starting at the upper edge of the liver and continuing until the pelvis. The intra-abdominal visceral fat volume was measured by drawing a line within the muscle wall surrounding the abdominal cavity. The abdominal subcutaneous fat volume was calculated by subtracting the visceral fat volume from the total abdominal fat volume. By comparison, the intra-abdominal visceral and subcutaneous fat areas were determined at the umbilical level by the established slice-by-slice CT scanning technique. RESULTS: V(vol) was correlated positively with visceral fat area (V(area)) measured by conventional CT in both genders (in men (n=79) V(vol) vs V(area), r=0.81 P<0.0001; in women (n=74) V(vol) vs V(area), r=0.85, P<0.0001). S(vol) also showed a positive correlation with subcutaneous fat area (S(area)) in both genders (in men (n=78) S(vol) vs S(area), r=0.95, P<0.0001; in women (n=74) S(vol) vs S(area), r=0.92, P<0.0001). CONCLUSION: We have reported a novel method for measuring intra-abdominal fat volume by the use of helical CT. PMID- 11896497 TI - Recent weight changes and weight cycling as predictors of subsequent two year weight change in a middle-aged cohort. AB - OBJECTIVE: To evaluate the influence of recent weight changes (weight gain, loss and cycling) on subsequent weight changes. DESIGN: Prospective cohort study with 2 y of follow-up. Data analysis with a polytomous logistic regression model. SUBJECTS: A total of 18 001 non-smoking subjects, 6689 men and 11 312 women, from the general population. MEASUREMENTS: Body height and weight measurements and interview data on lifestyle habits and medical history at baseline. For follow up, self-administered questionnaires for assessment of body weight and incident diseases. RESULTS: Recent changes in body weight, that is weight gain, weight loss and weight cycling, were significant predictors of subsequent weight changes in both men and women after controlling for age, baseline BMI and several lifestyle and behavioural characteristics as potential confounding factors. Weight cycling before baseline was the strongest predictor of subsequent large weight gain (> or =2 kg) with an odds ratio (OR) of 4.84 (95% confidence interval (CI) 3.34-7.02) in men. In women, prior weight loss was the strongest predictor of subsequent large weight gain (OR 4.77; 95% CI 3.63-6.03), followed by weight cycling (OR 3.02; 95% CI 2.15-4.25). CONCLUSION: These data indicate the need for thorough weight history assessment to identify those who are most likely to gain weight. Effective weight control before the development of obesity or after intentional weight loss due to obesity should be a primary goal in the management of obesity. PMID- 11896498 TI - Differential associations of body mass index and adiposity with all-cause mortality among men in the first and second National Health and Nutrition Examination Surveys (NHANES I and NHANES II) follow-up studies. AB - OBJECTIVE: The frequently observed U-shaped relationship between body mass index (BMI; kg/m(2)) and mortality rate may be due to the opposing effects of fat mass (FM) and fat-free mass (FFM) components of BMI on mortality rate. The purpose is to test the hypothesis stated above. DESIGN: Longitudinal prospective cohort studies. The mortality follow-up of the first and second National Health and Nutrition Examination Surveys (NHANES I and NHANES II). SUBJECTS: A total of 10 169 male subjects aged 25-75 who participated in NHANES I and II were selected for analyses. Follow-up continued until 1992. The mean follow-up time was 14.6 y for NHANES I and 12.9 y for NHANES II. Ninety-eight percent of the participants were successfully followed representing a total of 3722 deaths. MEASUREMENTS: Subscapular and triceps skinfolds thickness were used as FM indicators, whereas upper arm circumference was used as a FFM indicator. The Cox proportional hazards model tested the relationships of BMI, FM and FFM with all-cause mortality adjusting for age, smoking status, race and education levels. RESULTS: BMI had a U-shaped relationship with mortality, with a nadir of approximately 27 kg/m(2). However, when indicators of FM and FFM were added to the model, the relationship between BMI and mortality became more nearly monotonic increasing. Moreover, the relationship between FM indicator and mortality was monotonic increasing and the relationship between FFM indicator and mortality was monotonic decreasing. CONCLUSION: These results support the hypothesis that the apparently deleterious effects of marked thinness may be due to low FFM and that, over the observed range of the data, marked leanness (as opposed to thinness) has beneficial effects. PMID- 11896499 TI - Impact of overweight and obesity on health-related quality of life--a Swedish population study. AB - OBJECTIVE: To investigate the impact of overweight and obesity on health-related quality-of-life (HRQL) in the general population in western Sweden. DESIGN: Cross sectional survey. SUBJECTS: A total of 5633 men and women aged 16-64 y born in Sweden. MAIN OUTCOME MEASURES: Scale and summary component scores of the SF-36 Health Survey. RESULTS: Obese men aged 16-34 y rated their HRQL lower than normal weight men did on all four physical health scales of the SF-36 and on two of the four mental health scales. Obese women in the same age group rated their health worse than normal-weight women on three of the physical health scales. Thus, in younger men and women the analysis indicated a clearer negative association between obesity and physical health than between obesity and mental health. Obese women aged 35-64 y rated their health worse on all scales than normal-weight women did, while obese men in this age group rated their health worse on only two SF-36 subscales-physical functioning and general health perception. The massively obese men and women suffered from a poor level of HRQL. CONCLUSION: Not only does the level of obesity affect HRQL, the impact of overweight and obesity also differs by age and sex. The importance of aspects of both physical and mental health should be fully recognised. PMID- 11896500 TI - Obesity prevention: the case for action. PMID- 11896501 TI - Systolic vs diastolic blood pressure and the burden of hypertension. PMID- 11896502 TI - Is there an epidemic of primary aldosteronism? PMID- 11896503 TI - Is aldosterone the missing link in refractory hypertension?: aldosterone-to-renin ratio as a marker of inappropriate aldosterone activity. AB - Use of the random aldosterone-to-renin ratio (ARR) as a reliable marker of inappropriate aldosterone activity has led to primary aldosteronism (PA) being increasingly diagnosed in hypertensive patients. At least 10% of hypertensives have been found to have PA, the majority of whom presumably have bilateral adrenal hyperplasia or idiopathic hyperaldosteronism as an aetiology for PA. Whilst these patients clearly have excess aldosterone activity, they have in common many features that are found in hypertensive patients in general, amongst which include heightened angiotensin II adrenal sensitivity. Whether these individuals belong within the spectrum of 'essential hypertension' is being debated, but is probably irrelevant clinically since they appear to respond favourably to spironolactone treatment. In addition, there is recent evidence suggesting that these patients overexpress a key enzyme involved in aldosterone production, the aldosterone synthase, the activity of which appears to relate to its genotypic variation. PMID- 11896504 TI - Primary aldosteronism, a common entity? the myth persists. AB - Primary aldosterone excess or hyperaldosteronism is an important cause of hypertension which, when associated with an aldosterone secreting adenoma, is amenable to surgical cure. The biochemical hallmarks of the condition are a relative excess of aldosterone production with suppression of plasma levels of renin (a proxy for angiotensin II, the major trophic substance regulating aldosterone secretion). This combination of a high aldosterone and a low renin is however more commonly associated with 'nodular hyperplasia' of the adrenal glands, a condition not improved by surgery and variably responsive to the effects of the mineralocorticoid antagonist, spironolactone. Until recently the prevalence of either form of secondary hypertension has been thought to be low such that few clinicians 'hunted' for it in the absence of hypokalaemia (the traditional clue for the syndrome). This view has been challenged, firstly by the realisation that no more than 50% of such patients will have a low plasma potassium and secondly by the assumption that a 'normal' plasma aldosterone is in fact inappropriately elevated if the renin level is low. A single measurement of the ratio of aldosterone to renin levels is claimed to be highly predictive of patients who will have primary aldosterone excess. This paper examines the logic behind such claims and presents evidence from the literature that an abnormal ratio is simply a different description of the low renin state and that such patients do not necessarily have mineralocorticoid hypertension. Most patients 'discovered' by this test will have what many call low-renin hypertension, a condition not amenable to specific therapy. Claims that they are peculiarly sensitive to the hypotensive effects of spironolactone have not been tested in controlled trials. The test would however be expected to pick up those individuals with true Conn's syndrome but such patients remain too few in number to justify widespread use of an expensive screening test. PMID- 11896505 TI - Systolic vs diastolic blood pressure: community burden and impact on blood pressure staging. AB - Systolic blood pressure (SBP) is a more frequent cardiovascular risk factor than diastolic blood pressure (DBP), and has a greater impact on blood pressure staging, though this can vary with age, sex and country. Therefore this paper compares SBP and DBP in terms of community burden and impact on blood pressure staging, among Spain's middle-aged population. Data were drawn from a cross sectional study on a representative sample of the Spanish population aged 35-64 years. Blood pressure was determined under standardised conditions, and was classified as per WHO-ISH and JNC-VI criteria. Prevalence of SBP > or =140 mm Hg was 34.1%, and that of DBP > or =90 mm Hg, 30.9%. A total of 12% of subjects had isolated systolic hypertension (ISH) and 8.7% had isolated diastolic hypertension (IDH). Of treated hypertensives, 31% had their SBP controlled and 34% their DBP controlled. Of subjects not undergoing antihypertensive drug therapy, 60.8% had congruent SBP and DBP levels, 22.5% were up-staged on the basis of their SBP, and 16.7% were up-staged on the basis of their DBP. SBP alone thus correctly classified JNC-VI staging in 83.3% of subjects vs 77.5% for DBP alone. It was solely among the population >50 years of age, in both sexes, that systolic proved more frequent than diastolic hypertension, ISH greater than IDH prevalence, SBP worse than DBP control, and the percentage of SBP higher than that of DBP up staged subjects. SBP constitutes a greater community burden than does DBP, and has a greater impact on blood pressure staging in Spain's middle-aged population. However, the differential impact of SBP and DBP upon blood pressure burden and staging is favourable to SBP only among subjects >50 years old. These findings are in accordance with recent guidelines on hypertension management. PMID- 11896506 TI - Efficacy of eprosartan in combination with HCTZ in patients with essential hypertension. AB - This randomised, double-blind study was designed to investigate the efficacy of a once-daily (OD) combination of the AT(1) receptor blocker, eprosartan 600 mg, and the thiazide diuretic, hydrochlorothiazide (HCTZ) 12.5 mg, in patients with mild to moderate hypertension (sitting diastolic blood pressure (sitDBP) > or =98 mm Hg and < or =114 mm Hg) not adequately controlled with eprosartan 600 mg OD. A total of 494 patients entered the open-label monotherapy run-in phase, which consisted of eprosartan 600 mg OD for 3 weeks. Patients who responded to monotherapy were not eligible to enter the randomised phase of the study and were withdrawn. The remaining 309 patients were then randomised to either eprosartan 600 mg plus HCTZ 12.5 mg OD or to continue on eprosartan 600 mg OD. In the eprosartan plus HCTZ combination group, both sitDBP and sitting systolic blood pressure (sitSBP) were significantly reduced compared with the eprosartan monotherapy group. In addition, the response rate was higher in the combination group compared with the monotherapy group. There were no significant effects on reduction of sitDBP due to gender, prior use of antihypertensives or baseline severity of hypertension. The tolerability profile for the combination group was similar to that for the monotherapy group. Headache was the most frequent adverse event in both treatment groups. The majority of adverse events were mild to moderate in intensity. In this study of patients who were unresponsive to eprosartan monotherapy for 3 weeks, a combination product of eprosartan 600 mg and HCTZ 12.5 mg was shown to be an effective and well tolerated treatment. PMID- 11896507 TI - Ambulatory blood pressure characteristics in normotensive and treated hypertensive older people. AB - The objective of this study was to determine the normal values and characteristics of 24-h ambulatory blood pressure (ABP) and to describe the ABP level of treated hypertensive subjects in an older Finnish population. ABP was measured in 502 randomly selected subjects aged 64 years or over living in a Finnish municipality (mean age 70 years, range 64-87 years). A total of 211 subjects did not have blood pressure (BP) affecting medication. ABP measurements were taken every 30 min for 24 h, and the day- and night-time periods were diary based. The results were that in untreated subjects, the average office BP was 134/82 +/- 16/9 (s.d.) mm Hg for men and 140/81 +/- 18/8 mm Hg for women. The 24 h average BP was 120/75 +/- 14/8 mm Hg (95th percentile upper limit 145/93 mm Hg) for men and 125/75 +/- 15/7 (95th = 154/89 mm Hg) for women. The daytime averages were 127/78 +/- 12/7 mm Hg (95th = 154/99 mm Hg) and 131/78 +/- 15/7 mm Hg (95th = 158/91 mm Hg) for men and women, respectively. The ABP daytime value of 130/83 mm Hg corresponded best to the office BP value of 140/90 mm Hg. All BP values were significantly higher in the treated hypertensive group compared to the normotensive group. Night-time BP was markedly lower than daytime BP, and no difference in circadian variability was found between the normotensive and hypertensive subjects. Both office and ambulatory BPs were significantly higher in women than in men. This study provides sex-specific normal values for ABP in a 64 to 87-year-old age group. The normal values of ABP were markedly lower than the office BP values. Hypertensives, even when treated, tended to have elevated values. PMID- 11896509 TI - Circadian rhythm of blood pressure is transformed from a dipper to a non-dipper pattern in shift workers with hypertension. AB - Shift workers make great use of health care services because they are associated with increased cardiovascular morbidity and mortality. Whether the circadian rhythm of blood pressure rapidly adapts to shift work is controversial. It is unknown if shift work has adverse effects on blood pressure in patients with hypertension. To evaluate the effects of shift work, we examined 12 male shift workers with untreated hypertension aged 53.6 +/- 2.5 years. Twenty-four hour ambulatory blood pressure monitoring was performed three times as follows: the last day of a 4-day period of day shifts (09.00 to 21.00), the first day of a 4 day period of night shifts (21.00 to 09.00), and the fourth day of night shifts (21.00 to 09.00). Blood pressure at night-time dropped significantly in the day shift workers, showing a dipper pattern. Average differences in blood pressure in the sleep-wake cycle were decreased by 8.5% at the beginning of night shift work showing a non-dipper pattern. After 4 days the pattern was completely reversed to a dipper pattern. The results indicate that the circadian blood pressure pattern is changed from a dipper to a non-dipper pattern on the first day of the night shift and reverses to a dipper pattern within a few days. We suggest that night shift work may have unfavourable effects on blood pressure in patients with hypertension. PMID- 11896508 TI - Impact of an ACE inhibitor and calcium antagonist on microalbuminuria and lipid subfractions in type 2 diabetes: a randomised, multi-centre pilot study. AB - BACKGROUND: Microalbuminuria (MA) is associated with increased cardiovascular risk and lipid abnormalities in people with type 2 diabetes. ACE inhibitors and calcium channel blockers (CCBs) reduce MA and are neutral on total cholesterol and triglycerides. The effect of ACE inhibitors and CCBs on lipid subfractions such as Lp(a), apolipoprotein (apo) A1, apo B, and others, however, is unclear. The current study tests the hypothesis that a fixed-dose combination of an ACE inhibitor, benazepril (B) with the dihydropyridine CCB, amlodipine (A), will further reduce arterial pressure and reduce atherogenic lipid fractions compared to either agent alone. DESIGN: A multicentre, randomised, open-label, parallel group design was used to study 27 participants with type 2 diabetes. Measurements for total cholesterol, high- and low-density lipoprotein (HDL and LDL), triglycerides, apo A1, apo B, Lp(a), MA, arterial pressure and creatinine clearance were obtained at baseline and at 12-week intervals during the 36 week study. RESULTS: Arterial pressure was significantly reduced at 36 weeks in all three groups (P = 0.0078 for A, P = 0.0039 for B, and P = 0.0313 for A+B). MA was lowered in all groups with relatively greater reductions in the B (P < 0.05) and A+B groups (P < 0.03) vs A. An increase in mean HDL-cholesterol from baseline was noted in the B and A+B groups; P < 0.05), but not in the A group. A trend was also observed between the rise in HDL-cholesterol and the reduction in MA in the B and A+B groups. Additionally, only the B group exhibited a decrease in the median value of Lp(a) (P < 0.05). CONCLUSION: These data support the concept that ACE inhibition with B reduces the atherogenic profile by decreasing Lp(a) and increasing HDL-cholesterol, the latter being correlated with reductions in MA. While A+B exhibited similar trends in lipid subfractions and MA as B, this group had the greatest reduction in systolic blood pressure of the three groups. Thus, use of A+B offers the benefits of a decreased atherogenic profile with a higher probably of achieving goal blood pressure as recommended by national guidelines. PMID- 11896510 TI - Impact of calcium antagonists on bleeding time in patients with chronic renal failure. AB - Haemorrhagic diathesis develops in chronic renal failure, in which calcium antagonists are used widely as antihypertensive agents. Although calcium antagonists are reported to impair platelet function, it has not been examined whether calcium antagonists alter bleeding time. The present study was conducted to clarify whether calcium antagonists affect bleeding time in chronic renal failure. Patients with chronic renal failure without and with calcium antagonists were enrolled (n = 156), and bleeding time (Ivy's method) as well as blood parameters (BUN, creatinine, platelet counts, and haemoglobin) were compared in patients with normal and prolonged bleeding time. Among patients not taking calcium antagonists (n = 34), three cases manifested prolonged bleeding time, whereas abnormal bleeding time was observed in 31 patients out of 122. Positive correlations were observed between bleeding time and BUN in both calcium antagonist-untreated (r = 0.46) and -treated groups (r = 0.25). The odds ratio for prolongation of bleeding time in patients taking calcium antagonists was 3.52 (95% CI, 1.01-12.33). In 12 calcium antagonist-treated patients with prolonged bleeding time, the withdrawal of calcium antagonists markedly shortened bleeding time (from 11.3 +/- 0.8 to 5.4 +/- 0.8 min, P < 0.05, n = 12). In contrast, in the additional group (n = 9), the continued treatment with calcium antagonists had no effect on bleeding time (from 11.7 +/- 0.9 to 10.0 +/- 1.0 min). Despite the inhibitory effect of calcium antagonists on bleeding time, no clinically serious events associated with haemorrhagic diathesis developed. In conclusion, calcium antagonists prolong bleeding time in patients with chronic renal failure. The subclinical (laboratory) effect of calcium antagonists however is not necessarily associated with haemorrhagic events of clinical significance. PMID- 11896511 TI - Relation of urinary urea to blood pressure: interaction with urinary sodium. AB - A previous study reported that urinary markers of protein intake are inversely related to blood pressure via unknown mechanisms. In man and rats, protein intake affects renal function and increases renal sodium excretion. The present study investigates the relation between markers of protein intake and blood pressure and the possible role of sodium in this relation. Blood pressure status, overnight urinary urea as index of protein intake, urinary and plasma sodium, and other variables were measured in a population sample of 3705 men and women, aged 25-74 years, without high plasma creatinine. Urinary urea was inversely related to blood pressure and hypertension: in multivariate analyses, 6.5 mmol/h higher urinary urea (about one s.d. in men and women) was related to 4.25 mm Hg lower systolic blood pressure (95% confidence interval = 1.34-8.49), and to 0.65 lower risk of hypertension (95% CI 0.34-0.87). An interaction was found between overnight urinary sodium and the relation of urinary urea to blood pressure: the relation was significant only in persons with overnight urinary sodium above the median. Urinary urea was significantly and inversely also related to plasma sodium. Data confirm an inverse relation to blood pressure of protein intake as measured by urinary urea. The possibility of sodium-related mechanisms is supported by the interaction of urinary sodium with the relation and by the inverse association of urinary urea with plasma sodium. The hypothesis is made that high protein intake could counteract sodium-dependent blood pressure rise via stimulation of renal sodium excretion. PMID- 11896512 TI - More reasons to hold on additional doses of antenatal steroids. PMID- 11896513 TI - Multiple courses of antenatal corticosteroids are associated with early severe lung disease in preterm neonates. AB - OBJECTIVE: Determine whether the increased neonatal mortality following repeated courses of antenatal corticosteroids (ANCS), observed in the Thyrotropin Releasing Hormone (TRH) Trial, was related to confounding maternal risk factors or specific preterm morbidities. STUDY DESIGN: A post hoc analysis of 595 TRH trial neonates, 26 to 32 weeks' gestation, studied the association between > or =3 courses ANCS and mortality. Potential confounding maternal factors and preterm morbidities were evaluated using logistic regression and log likelihood modeling. RESULTS: Mortality was 9.2% after > or =3 courses (13/141) vs. 4.8% after 1 or 2 courses (22/454). This association was not explained by maternal factors, or other common preterm morbidities. However, 15/141 infants receiving > or =3 courses (10.6%) had early severe lung disease (ESLD) with 10 deaths, compared to 16/454 of the 1- to 2-course infants (3.5%) with 7 deaths (odds ratio 3.5, p<0.001). CONCLUSIONS: ESLD, but not maternal risk factors, was associated with increased mortality in preterm infants after > or =3 courses ANCS. PMID- 11896514 TI - Effect of recombinant erythropoietin on "late" transfusions in the neonatal intensive care unit: a meta-analysis. AB - OBJECTIVES: Using the approach of a meta-analysis, we sought to determine whether the administration of recombinant erythropoietin (rEpo) to very low birth weight (VLBW) infants, after the first week of life, results in fewer "late" transfusions. STUDY DESIGN AND METHODS: The guidelines set forth by the Cochrane Neonatal Review Group were used to identify all relevant studies. Medline was searched from January 1990 to November of 2000. Studies that used a randomized, placebo-controlled, and double-masked design were deemed acceptable. RESULTS: Eight studies meet the inclusion criteria. These involved 357 VLBW neonates: 183 rEpo and 174 placebo recipients. The neonates in the rEpo group received fewer erythrocyte transfusions during the study period than did those in the placebo group; the common odds ratio (OR)=0.33; 95% confidence interval (CI) 0.21-0.51. Furthermore, the rEpo effect size was a function of the dose of rEpo administered (p=0.0001). CONCLUSION: A meta-analysis of the most scientifically rigorous studies on this topic indicates that administration of rEpo to VLBW infants reduces "late" erythrocyte transfusions in a dose-dependent manner. PMID- 11896515 TI - The effects of the introduction of a high-nutrient transitional formula on growth and development of very-low-birth-weight infants. AB - OBJECTIVE: The purpose of this study was to determine the impact of introducing a high-nutrient transitional formula (TF) for use after discharge on the growth and development of premature infants. STUDY DESIGN AND METHODS: This was a cohort study of all surviving infants with a birth weight < or =1250 g admitted to the neonatal intensive care unit between January 1, 1995 and December 31, 1996. Infants with major congenital abnormalities were excluded. There were 180 infants discharged, including 66 on TF and 114 on standard formulas for full-term infants. RESULTS: Use of TF started the week before discharge, and increased from 10% in 1995 to 66% in 1996 (p<0.001). Regression analyses controlling for multiple confounders identified TF as a significant contributor to improved weight at 3 months and length at 18 months. Bayley developmental scores were not affected. CONCLUSION: Introduction of a TF for very-low-birth-weight infants resulted in improved growth after discharge. PMID- 11896516 TI - Newborn hearing screening: costs of establishing a program. AB - OBJECTIVE: To evaluate the costs and performance characteristics associated with the start-up phase of Universal Newborn Hearing Screening Programs, one utilizing automated auditory brainstem response (AABR) and the other using transient evoked otoacoustic emissions (TEOAE). STUDY DESIGN: Economic and performance data were collected at the initiation of both screening programs. Data were collected until 1500 newborn infants were screened or until a referral rate for further audiologic evaluation at hospital discharge of less than or equal to 5% was achieved. Data collected included screening pass/fail rates, referral rates and personnel, equipment, and supply utilization. Actual costs of personnel, equipment, and supplies were used. Statistical comparisons of proportions using z statistic with the one-tailed test and an alpha of 0.01 were made. RESULTS: Screening in the AABR program was performed by neonatal nurses, whereas screening in the TEOAE program was performed by master's level audiologists. The average age at initial screen was 29 hours for TEOAE, and 9.5 hours for AABR. Eighty-four percent of infants was screened within 24 hours in the AABR program, in contrast to 35% in the TEOAE program. Throughout the duration of the study, the referral rate at hospital discharge remained approximately 15% for the TEOAE program. The AABR referral rate began at 8% and was less than 4% at the completion of the study. Pre-discharge total costs for initiating and establishing the programs were US$49,316 for TEOAE and US$47,553 for AABR. Cost per infant screened was US$32.23 and US$33.68, respectively. When post-discharge screening and diagnostic evaluation costs were included, the total cost per infant screened was US$58.07 for TEOAE and US$45.85 for AABR. CONCLUSION: AABR appears to be the preferred method for universal newborn hearing screening. AABR was associated with the lowest costs, achieved the lowest referral rates at hospital discharge, and had the quickest learning curve to achieve those rates. PMID- 11896517 TI - Risk score for antenatal bacterial vaginosis: BV PIN points. AB - OBJECTIVE: Develop a clinical risk score to screen for antenatal bacterial vaginosis (BV), irrespective of symptoms. STUDY DESIGN: Cohort study of 913 pregnant women with last menstrual periods between January 30, 1995 and February 22, 1997. BV was evaluated by Nugent-scored vaginal smears (scores of 7 to 10 considered positive) between 24 and 29 weeks' gestation. Forty-four potential risk factors were assessed. RESULTS: 17.8% of women had BV, of whom 22% were screened for BV by the usual care provider. Logistic regression-adjusted analyses found six predictors: vaginal pH>4.5 (OR=11.6, 95% confidence interval [CI] [7.8, 17.2]); black race (OR=1.9, 95% CI [1.3, 2.8]); condom use during pregnancy (OR=1.6, 95% CI [1.0, 2.5]); antenatal BV (OR=1.7, 95% CI [1.0, 2.8]); absence of sperm on smear (OR=1.7, 95% CI [1.0, 2.9]); and no history of sexually transmitted diseases (OR=1.6, 95% CI [1.0, 2.5]). Risk score weights were 5 for an elevated vaginal pH and 1 otherwise. The sensitivity and specificity of screening women with scores > or =4 were both 77%; this would involve screening 33% of patients. CONCLUSION: Approximately 80% of our BV cases were asymptomatic, emphasizing the need for objective risk assessment. Using six factors, clinicians can identify pregnant women at risk for BV. PMID- 11896518 TI - Fetal gender and cocaine exposure as determinants of cord blood gamma-glutamyl transferase activity. AB - OBJECTIVE: The serum activity of the hepatic enzyme gamma-glutamyl transferase (GGT) is elevated in the newborn relative to older age groups. Few reports to date have studied the influence of perinatal factors on neonatal serum GGT and no study has assessed the influence of maternal drug ingestion. STUDY DESIGN: Cord blood was randomly collected from 234 liveborn infants and correlated with a range of maternal and fetal perinatal variables to assess influences on cord blood GGT. RESULTS: Our study showed that the range of cord blood GGT activity in 234 randomly selected term newborns was 22 to 556 IU/l. In a subgroup of 75 newborns, GGT activity was independently influenced by only two of the variables studied - cocaine exposure and fetal gender (p=0.009, r=0.39). Females had a lower GGT than males (95+/-66 vs 130+/-90 IU/l, p<0.001) while GGT activity in cocaine-exposed newborns was lower than in cocaine-nonexposed newborns (96+/-48 vs 142+/-109, p<0.01). Birth weight, race, gestational age, and maternal serum GGT were not found to significantly influence cord blood GGT activity. Maternal GGT was uniformly normal and was not affected by any of the variables tested. CONCLUSION: Our findings demonstrate that the reference range for cord blood GGT activity is wide and appears to be influenced by fetal gender and cocaine exposure. PMID- 11896519 TI - Effect of an evidence-based hand washing policy on hand washing rates and false positive coagulase negative staphylococcus blood and cerebrospinal fluid culture rates in a level III NICU. AB - OBJECTIVE: To determine the effect of implementing an evidence-based hand washing policy on between-patient hand washing compliance and on blood and cerebrospinal fluid (CSF) culture rates in a level III neonatal intensive care unit (NICU). METHODS: An evidence-based hand washing policy, supported by an intensive education program, was introduced in a regional NICU. A total of 2009 preintervention neonates (16,168 patient days) over 17 months were compared to 676 postintervention neonates (5779 patient days) over 6 months. Hand washing compliance and rates of blood and CSF cultures yielding coagulase negative staphylococci (CONS) were compared before and after intervention. RESULTS: Compliance with appropriate between-patient hand washing improved (from 47.4% to 85.4%, p=0.001) after the hand washing policy was introduced. The rate of cultures positive for CONS declined from 6.1+/-2.3 to 3.2+/-1.6 per 1000 patient days (p=0.005). Most of this reduction was attributable to a reduction in false positive cultures, from 4.2+/-2.4 to 1.9+/-1.8 per 1000 patient days (p=0.042), but there was a trend toward decreased true-positive cultures (from 2.1+/-1.2 to 1.2+/-1.0 per 1000 patient days, p=0.074) as well. Potential confounders and demographics factors were similar between the control and intervention subjects. CONCLUSION: Implementation of an evidence-based hand washing policy resulted in a significant increase in hand washing compliance and a significant decrease in false-positive coagulase negative staphylococcal blood and CSF culture rates. Exploratory data analysis revealed a possible effect on true-positive coagulase negative staphylococcal blood and CSF culture rates, but these results need to be confirmed in future studies. PMID- 11896520 TI - QTc interval in infants receiving cisapride. AB - OBJECTIVE: To examine the effect of cisapride on the corrected QT (QTc) interval in infants over a 14-day period. STUDY DESIGN: A prospective cohort study of infants receiving cisapride (0.8 mg/kg per day). Twelve-lead electrocardiograms were obtained before and 3, 5, 7, and 14 days after cisapride initiation. RESULTS: Fifty infants completed the study; none had arrhythmias. Fifteen of 50 infants (30%) developed QTc interval > or =450 msec; QTc interval normalized in 13 of 15 infants. Infants with QTc interval on day 3 > or =2 standard deviations above the mean baseline QTc interval (401+40 msec) were more likely to develop prolonged QTc interval (p<0.0001). CONCLUSION: QTc interval prolongation was noted in 30% of infants. Subsequently, the majority of those infants had QTc interval normalization by day 14 of cisapride therapy. QTc interval 3 days following cisapride initiation may identify infants at risk for transient QTc interval prolongation. With appropriate monitoring, hospitalized infants receiving cisapride may have improved gastrointestinal motility without cardiac morbidity. PMID- 11896521 TI - Uterine contraction pattern as a predictor of the mode of delivery. AB - OBJECTIVE: To determine whether analysis of the frequency or regularity of uterine contractions can distinguish between women delivering vaginally and women requiring Cesarean section for dystocia. STUDY DESIGN: This was a case-control study. Cases meeting the following criteria were identified: nulliparous women in spontaneous labor, singleton pregnancy, cephalic presentation, 37 to 42 weeks' gestation, Cesarean section for dystocia, a minimum of three analyzable hours of electronically archived cardiotocograph (CTG) prior to onset of the second stage. Cases delivered by Cesarean section for dystocia were each matched with two controls delivering vaginally. The peak of each contraction was visually identified and electronically marked on each CTG trace. A moving average (MTIME) and standard deviation (SDTIME) of five interpeak times were calculated for successive 30-minute periods and plotted against cervical dilatation for each group. RESULTS: Overall, both MTIME and SDTIME fell as labor progressed, with these changes being more marked in the vaginal delivery group. In women requiring oxytocin, the rate of fall of MTIME and SDTIME was significantly greater following oxytocin in the group which subsequently delivered vaginally (slope of MTIME -2.71 pre-oxytocin and -28.95 post-oxytocin, p=0.0004; slope of SDTIME 0.44 and -6.44, p=0.0002). No such change was seen in the Cesarean section group. CONCLUSION: As normal labor progresses, there is a shortening of the intercontraction interval and an increase in contraction regularity. A successful response to oxytocin augmentation may be predicted by the change in contraction pattern following treatment. PMID- 11896522 TI - Comparison of capillary blood sampling using an automated incision device with and without warming the heel. AB - OBJECTIVE: To determine if heel warming prior to heelstick increases the volume of blood collected compared to no warming. STUDY DESIGN: Randomized controlled trial involving 100 preterm and term infants requiring capillary blood sampling (CBS), randomized to receive warming (Control) (n=50) or nonwarming (Experimental) (n=50). RESULTS: This sample of infants showed no benefit of warming the heel prior to CBS. Volume of blood, collection time, crying time, and repeat procedures were not different between groups. More infants in the Control group received squeezing during the procedure compared to the Experimental group. CONCLUSION: Heel warming prior to CBS may be an unnecessary technique in preterm and term infants that expends nursing time and hospital financial cost. PMID- 11896523 TI - Tuberculosis skin testing among HIV-infected pregnant women in Miami, 1995 to 1996. AB - OBJECTIVE: Approximately 6000 women deliver annually at Jackson Memorial Hospital in Miami, where 2.4% of women has human immunodeficiency virus (HIV) and 60% is foreign-born. We conducted a retrospective review of prenatal records among HIV infected women to evaluate tuberculin skin testing (TST). STUDY DESIGN: We determined how many women had TSTs placed and read, and the TST results. RESULTS: We identified 207 HIV-infected women, 87% of such women delivering in 1995 to 1996. Most did not know their HIV status before seeking prenatal care (109, 54%) and most (176, 85%) had TSTs done. Of the women, 45 had positive TSTs, 96 had negative TSTs, and 35 were anergic. Most results were not recorded using millimeters of induration. Two women (1%) had active tuberculosis (TB) disease. CONCLUSION: Overall, 21% of all HIV-infected women had positive TSTs and 1% had active TB disease. Focused TB skin testing should be part of routine prenatal care in clinics serving populations at high-risk for TB, such as those with HIV infection and the foreign-born. PMID- 11896524 TI - Effect on sleep position on apnea and bradycardia in high-risk infants. PMID- 11896525 TI - Effect of home monitoring on a high-risk population. AB - A large cohort of infants (8,998) at high risk for sudden and unexpected death was followed with home cardiorespiratory monitoring over a five-year period. These infants included premature infants (23-36 weeks post-conceptual age), SIDS siblings, and infants who experienced an Apparent Life-Threatening Event. The overall SIDS rate in this high-risk population was 0.55/1,000, a rate significantly less than the 0.85 deaths/1,000 reported in the "general population" of Georgia over this same time period. In addition, we report our experience with using home monitors as a diagnostic tool, as well as how monitors can actually be cost-effective. Editorial opinions, and lay press summaries of the CHIME study (JAMA, May 2, 2001) imply that home cardiorespirtory monitors are of little value. Despite the fact that the study never made this claim, many clinicians are now referring to this study as evidence that home monitoring is ineffective and not needed. This article disputes those misconceptions about home cardiorespiratory monitors based on our experience with a large high-risk population of infants. PMID- 11896526 TI - Caudal regression syndrome versus sirenomelia: a case report. AB - We describe a newborn with clinical features of sirenomelia including fused lower limbs with medial position, absence of fibula, anal atresia, bilateral renal agenesis, and a single large umbilical artery. Recent literature describing the etiology of sirenomelia and relationship to caudal regression syndrome is reviewed. PMID- 11896527 TI - Caudal Regression Syndrome in twin pregnancy with type II diabetes. AB - Caudal Regression Syndrome (CRS) is a rare fetal complication of diabetic pregnancy, which can result in long-term neurological, urologic, and orthopedic complications. Although the exact teratogenic mechanism is not known, hyperglycemia appears to play a crucial role as a teratogen, and therefore, stringent control of diabetes preconceptually and in early pregnancy is presumed to reduce the risk of occurrence. We report an unusual case of CRS affecting only one of a set of monozygotic twins, suggesting that as yet, unidentified factors other than hyperglycemia are included in its causation. PMID- 11896528 TI - Neonatal sinovenous thrombosis associated with homozygous thermolabile methylenetetrahydrofolate reductase in both mother and infant. AB - The C677T mutation in 5,10-methylenetetrahydrofolate reductase (MTHFR) predicts substitution of valine for alanine at residue 223 (A223V). This thermolabile form of MTHFR has 50% reduced activity, has been associated with hyperhomocystinemia, and is a described risk factor for thrombosis in adults.(1-3) In addition, it has been associated with birth defects in the infants of affected mothers and with recurrent fetal losses.(4-6) We report the occurrence of sinovenous thrombosis in a newborn infant who presented with seizures. Both infant and mother were subsequently identified as having homozygous C677T alleles for MTHFR. PMID- 11896529 TI - Congenital ductus arteriosus aneurysm presenting with stridor in a newborn. PMID- 11896531 TI - Antiproliferative effect of plant cytokinin analogues with an inhibitory activity on cyclin-dependent kinases. AB - In this study, analogues of olomoucine, a previously described plant cytokinin analogue with cyclin-dependent kinase (CDK) inhibitory activity, were investigated for effect on CDK1 and CDK2 and for effect on cell proliferation. Eight new compounds exhibit stronger inhibitory activity on CDK1 and CDK2 and on cell proliferation than olomoucine. Some active compounds showed low inhibition of proliferation of normal myeloid growth. Improvement of inhibitory activity of known compounds with a C6-benzylamino group was brought about by substitution with one hydroxyl. Also, new C2 substituents associated with inhibitory activity on CDK and on cell proliferation are described. There was a significant correlation between effect on CDK and antiproliferative effect on the KG1 and Molt3 cell lines and on primary human lymphocytes, strongly suggesting that at least part of the antiproliferative effect of cytokinin analogues was due to inhibition of CDK activity. Cytokinin analogues induced apoptosis in a time- and concentration-dependent manner and changes in cell cycle distribution. The antiproliferative and pro-apoptotic effects of plant cytokinin analogues suggest that they are a new class of cytostatic agents and that they may find an application in the chemotherapy of cancer. PMID- 11896532 TI - New drugs for therapy of AML. PMID- 11896533 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF) priming with successive concomitant low-dose Ara-C for elderly patients with secondary/refractory acute myeloid leukemia or advanced myelodysplastic syndrome. AB - Patients with advanced MDS and secondary AML respond poorly to chemotherapy. Granulocyte-macrophage colony-stimulating factor (GM-CSF) can stimulate proliferation of leukemic blasts and sensitize these cells to the cytotoxic effects of S-phase-specific drugs. This is the first report of safety and efficacy of GM-CSF prior to and during cytarabine in a low-dose, intermittent regimen for elderly patients with poor risk acute myelogenous leukemia or myelodysplastic syndrome. Twenty patients, age 68 to 86 years, each received 250 microg/m2 of GM-CSF (Sargramostatin; Immunex, Seattle, WA, USA) subcutaneously (s.c.) or intravenously (i.v.) for 3 days followed by GM-CSF at the same dose and cytarabine 100 mg/m2 i.v. for 3 days. GM-CSF and cytarabine were both administered for 3 days during weeks 2 and 3 followed by a 3-week rest period. Rates of CR and PR were 20% and 40%, respectively. These included clinically significant resolution of cytopenias and transfusion requirements. Many of the responding patients had been heavily pretreated prior to enrollment. One- and 2 year survival estimates are 44% and 19%, respectively. Myelosuppression was the most significant toxicity. Our findings suggest that this novel combination of GM CSF with sequential and concomitant low-dose cytarabine can benefit patients with poor risk myeloid malignancies. PMID- 11896534 TI - The enhanced in vitro hematopoietic activity of leridistim, a chimeric dual G-CSF and IL-3 receptor agonist. AB - The in vitro activity of leridistim was characterized for cell proliferation, generation of colony-forming units (CFU) and differentiation of CD34+ cells. In AML-193.1.3 cells, leridistim exhibited a significant increase in potency compared to rhG-CSF, SC-65303 (an IL-3 receptor agonist) or an equimolar combination of rhG-CSF and SC-65303. CFU-GM assays demonstrated that at 50% of the maximum response, the relative potency of leridistim was 12-fold greater than the combination of rhG-CSF and rhIL-3 and 44-fold more potent than rhG-CSF alone. In multi-lineage CFU assays, a combination of erythropoietin (rhEPO) and leridistim resulted in greater numbers of BFU-E, CFU-GEMM and CFU-Mk than rhEPO alone. Ex vivo culture of peripheral blood or bone marrow CD34+ cells with leridistim substantially increased total viable cells over cultures stimulated with rhG-CSF, SC-65303, or a combination of rhG-CSF and SC-65303. Culture with leridistim, resulted in a greater increase in myeloid (CD15+/CD11b+), monocytic (CD41-/CD14+) and megakaryocytic (CD41+/CD14-) precursor cells without depleting the progenitor pool (CD34+/CD15-/CD11b-). These results demonstrate that leridistim is a more potent stimulator of hematopoietic proliferation and differentiation than the single receptor agonists (rhG-CSF and SC-65303) either alone or combined. These unique attributes suggest that leridistim may enhance hematopoietic reconstitution following myelosuppressive chemotherapy. PMID- 11896536 TI - Apoptosis induction by hypercross-linking of the surface antigen CD5 with anti CD5 monoclonal antibodies in B cell chronic lymphocytic leukemia. AB - We evaluated cells from 24 patients with B cell chronic lymphocytic leukemia (B CLL) to determine apoptosis induced by CD5 hypercross-linking. Following the CD5 hypercross-linking with anti-CD5 monoclonal antibodies (MoAbs), we identified 10 patients where CD5 hypercross-linking induced apoptosis (group A) and 14 patients whose cells were resistant to the anti-CD5 MoAbs (group B). The programmed cell death pathway of the cells from patient group A was caspase-3 and poly (ADP ribose) polymerase (PARP)-dependent, involved a reduction of the mitochondrial transmembrane potential DeltaPsi and a down-regulation of the anti-apoptotic Bcl 2, Mcl-1 and iNOS proteins. Early activation-associated molecules such as CD25 and CD69 were expressed at higher levels than in controls after 6 h of culture with anti-CD5 MoAb. The expression of CD5 and of CD72, the ligand for CD5, were significantly lower in group A compared with group B. Anti-CD20 MoAb had similar activity with anti-CD5 MoAb and the combination of the two MoAbs seemed to be additive. In this study, it is suggested that the cells from some B-CLL patients can be induced into programmed cell death by CD5 hypercross-linking with anti-CD5 MoAbs. PMID- 11896535 TI - Cell cycle progression of chronic lymphocytic leukemia cells is controlled by cyclin D2, cyclin D3, cyclin-dependent kinase (cdk) 4 and the cdk inhibitor p27. AB - B-CLL cells are arrested in G0/early G1 phase of the cell cycle and are characterized by a marked hyporesponsiveness towards a variety of polyclonal B cell activators. We have previously demonstrated that costimulation with CpG-ODN and IL-2 can overcome this proliferative defect. Cyclin D3 is the principal D type cyclin which mediates G1 progression in normal B cells, but in B-CLL cells both cyclin D2 and cyclin D3, were strongly upregulated upon stimulation. Both cyclins were associated with cdk4 but not with cdk6, which is the catalytic partner of D-type cyclins in normal B cells. Moreover, immune complexes consisting of cyclin D2 and cdk4 or cyclin D3 and cdk4 were both functional and phosphorylated the RB protein in vitro. The cell cycle inhibitor p27 plays a pivotal role in cell cycle progression of B lymphocytes and has been shown to be overexpressed in B-CLL cells. P27 was rapidly downregulated in B-CLL cells even when stimulated with a non-CpG-ODN or IL-2 alone, while only moderate regulation could be observed in normal B cells. Taken together, our findings demonstrate that regulation of early cell cycle progression differs between B-CLL cells and normal B cells. These findings do not only contribute to the understanding of B CLL pathophysiology, but might ultimately lead to the identification of new therapeutic targets. PMID- 11896537 TI - Molecular cytogenetic analysis of 10;11 rearrangements in acute myeloid leukemia. AB - MLLT10 (previously called AF10) is a moderately common MLL fusion partner predominantly occurring in acute monoblastic leukemia (AML-M5). 10;11 rearrangements require at least three breaks in order to generate an in-frame MLL MLLT10 fusion as a result of the opposite orientations of both genes on the respective chromosome arms. In this study, we describe a detailed molecular cytogenetic analysis of MLL-MLLT10 positive 10;11 rearrangements in two patients. We observed an as yet unreported chromosomal mechanism with at least four breakpoints, leading to MLL-MLLT10 gene fusion in a 24-year-old male. An inversion of 11q13-q23 with a breakpoint in the MLL gene was followed by an additional break 3' of MLL prior to insertion of the 11q segment into MLLT10. In a second patient, a 37-year-old male with AML-M5b, molecular cytogenetic analysis of an apparent 10;11 reciprocal translocation showed an intrachromosomal inversion of 3'MLLT10followed by a reciprocal translocation between 10p12 and 11q23. Review of the literature showed that all cases were the result of an inversion of either 10p or 11q followed by translocation 10p;11q or insertion of the inverted segment into MLLT10 or MLL. PMID- 11896538 TI - The AFT024 stromal cell line supports long-term ex vivo maintenance of engrafting multipotent human hematopoietic progenitors. AB - The immortalized murine stromal cell line AFT024 has been reported to maintain human hematopoietic progenitors in an undifferentiated state in vitro. In the current studies the beige/nude/xid (bnx) mouse in vivo xenograft model was used to examine the engraftment and multilineage generative potential of human hematopoietic progenitors after 2-3 weeks growth on AFT024 stroma, in comparison to primary stromal monolayers derived from post-natal human bone marrow. Eight to 12 months after transplantation of human CD34+CD38- cells from umbilical cord blood, cultured on AFT024 vs human stroma for 2-3 weeks, the murine bone marrow was harvested and analyzed for the presence of human myeloid and lymphoid cells. The mean percent engraftment of total human hematopoietic cells in the murine marrow was significantly higher after co-cultivation on AFT024 than on human stroma. Human myeloid and lymphoid lineage cells were detected in all mice. However, engraftment of myeloid lineage cells (CD33+), B lymphoid (CD19+), and T lymphoid cells (CD4+and CD8+) were significantly higher after co-cultivation of the human cells on AFT024 than on human stroma, prior to transplantation. Interestingly, the length of time in culture did not significantly affect the engraftment of the myeloid and T lymphoid lineage progenitors, but the percentage of B lymphoid lineage engraftment decreased significantly between 2 and 3 weeks of co-cultivation on both types of stroma. Cells with a primitive phenotype (CD45+/CD34-/CD38- and CD45+/CD34-/lin-) and cells with the capacity to generate secondary human CFU after recovery from the bnx bone marrow were maintained at significantly higher levels during culture on AFT024 stroma than on human stroma. The current studies demonstrate that the AFT024 murine stromal cell line supports the ex vivo survival and maintenance of human hematopoietic progenitors that are capable of long-term multilineage reconstitution for 2-3 weeks ex vivo, to levels superior to those that can be obtained using human stromal cells. PMID- 11896539 TI - Acquired skewing of Lyonization remains stable for a prolonged period in healthy blood donors. AB - The pattern of X-chromosome inactivation (XCIP), or Lyonization, can be used to distinguish monoclonal from polyclonal cell populations in females. However, a skewed XCIP exists in hematopoietic cells in approximately 40% of healthy elderly females, interfering with interpretation of clonality assays. In hematopoiesis, an active stem cell pool is assumed to be present within a larger population of inactive stem cells, with a continuous exchange of cells between the two compartments. The assumption that the active stem cell pool size decreases with age may explain the phenomenon of acquired skewing occurring by chance and predicts the XCIP of this population to fluctuate. This fluctuation should be reflected in the XCIP of peripheral granulocytes. We examined the XCIP for fluctuations in time in peripheral granulocytes, monocytes and T cells of young, middle-aged and elderly healthy females. We used an optimized HUMARA PCR assay that eliminates unbalanced DNA amplification. We found no fluctuations in XCIP in any age group in up to 18 months follow-up. We conclude that acquired skewing arises gradually in life without fluctuations in XCIP and that analysis at multiple time points cannot distinguish monoclonal hematopoiesis from normal, skewed hematopoiesis. PMID- 11896540 TI - Fusion gene transcripts and Ig/TCR gene rearrangements are complementary but infrequent targets for PCR-based detection of minimal residual disease in acute myeloid leukemia. AB - PCR-based monitoring of minimal residual disease (MRD) in acute leukemias can be achieved via detection of fusion gene transcripts of chromosome aberrations or detection of immunoglobulin (Ig) and T cell receptor (TCR) gene rearrangements. We wished to assess whether both PCR targets are complementary in acute myeloid leukemia (AML). We investigated 105 consecutive AML cases for the presence of fusion gene transcripts by reverse transcriptase polymerase chain reaction (RT PCR): AML1-ETO associated with t(8;21), CBFB-MYH11 with inv(16), PML-RARA with t(15;17), BCR-ABL with t(9;22), and MLL-AF4 with t(4;11). In 17 out of 105 AML cases (16%), fusion gene transcripts were found. Ninety-five of these AML patients (13 with fusion gene transcripts) were also investigated for the presence of IGH, IGK, TCRG and TCRD rearrangements by Southern blot and/or PCR heteroduplex analysis and sequencing. In nine out of 95 patients (9.5%), such rearrangements were found. Combined data revealed that only one patient with a fusion gene transcript had a coexistent Ig/TCR rearrangement. The nine AML patients with Ig/TCR rearrangements, as well as five additional AML patients from a previous study were investigated in more detail, revealing that Ig/TCR rearrangements in AML are immature and unusual. The presence of Ig/TCR rearrangements in AML did not correlate with RAG gene expression levels as determined by real-time quantitative PCR. In conclusion, fusion gene transcripts and Ig/TCR rearrangements are infrequent, but complementary MRD-PCR targets in AML. PMID- 11896541 TI - Inhibitors of angiogenesis selectively reduce the malignant cell load in rodent models of human myeloid leukemias. AB - Angiogenesis is essential for growth and metastasis of solid tumors and probably also for hematological malignancies. Angiogenic inhibitors, like endostatin (ES) and PI-88, retard cancer growth. We tested these in mice with juvenile myelomonocytic leukemia (JMML), and in rats with acute myeloid leukemia (BNML). Eight weeks after transplantation and with a continuous drug treatment for the last 4 weeks, the leukemic cell mass decreased from almost 90% of all bone marrow cells to about 15 and 45% with ES, to about 35 and 55% with PI-88, and to about 10 and 25% with ES + PI-88 in the leukemic mice and rats, respectively. The numbers of normal human bone marrow cells transplanted into mice were unchanged by the treatments. The microvessel density in leukemic animals given ES or PI-88 was 10-50% of that in untreated animals. Notably, simultaneous treatment with ES and PI-88 led to a reduction of about 95% in JMML mice and 85% in BNML rats. In vitro proliferation of either JMML or BNML cells was not significantly altered by either drug, demonstrating the selectivity of ES and PI-88 as angiogenic inhibitors. In conclusion, anti-angiogenic therapy may be a valuable adjunct to conventional treatment of leukemia. PMID- 11896542 TI - IL-1beta expression in IgM monoclonal gammopathy and its relationship to multiple myeloma. AB - We have shown that IL-1beta is not detectable in normal plasma cells but is produced by plasma cells from virtually all patients with multiple myeloma (MM). To extend our earlier work, IL-1beta expression was determined in 13 newly diagnosed patients with IgM monoclonal gammopathy. Eleven patients with Waldenstrom's macroglobulinemia (WM) and two patients with IgM MM were investigated for IL-1beta expression by in situ hybridization (ISH). All patients with WM had bone marrow biopsies consistent with the diagnosis, an IgM M-protein in the serum, and subsequently required chemotherapy. Seven of 11 patients with WM had an M-protein >3 g/dl and five patients had bone surveys performed that were negative for osteolytic disease. Two patients were diagnosed with IgM MM because of the presence of significant osteolytic disease on a metastatic bone survey. ISH for kappa, lambda, and IL-1beta expression was performed on bone marrow aspirates from each of the 13 patients. None of the neoplastic cells from the 11 patients with WM showed detectable IL-1beta expression by ISH. However, the neoplastic cells from both patients with IgM MM expressed IL-1beta mRNA at high levels. This aberrant IL-1beta production may explain the presence of bone lesions in the patients with IgM MM. PMID- 11896543 TI - Real-time quantitative PCR assays for deoxycytidine kinase, deoxyguanosine kinase and 5'-nucleotidase mRNA measurement in cell lines and in patients with leukemia. AB - The relative levels of the deoxycytidine kinase (dCK), deoxyguanosine kinase (dGK), and the 5'-nucleotidase (5'-NT) are of importance for the effect of many nucleoside analogues used in the treatment of hematological malignancies. To elucidate dCK, dGK and 5'-NT gene expressions in cell lines and in samples from patients with leukemia, we have established a real-time quantitative PCR (RQ-PCR) method. From the available dCK, dGK and 5'-NT cDNA sequences we designed specific primers and fluorogenic probes for the respective genes. The mRNA of dCK, dGK and 5'-NT was also measured by semi-quantitative RT-PCR, the enzyme activities by a radioactive substrate-based technique and Western blot was used to measure the amount of dCK and dGK protein. A MOLT-4 wild-type and its 9-beta-D arabinofuranosylguanine (Ara-G)-resistant subline was used for the methods comparisons and the RQ-PCR assay was used in 35 samples from pediatric patients with ALL and AML. The results from RQ-PCR for the cell lines were in agreement with the semi-quantitative RT-PCR. The mRNA expression for dCK, dGK and 5'-NT (expressed as the ratio of the respective gene and the reference gene) in pediatric ALL and AML patients showed a large interindividual variability from 0.06 to 2.34, non-detectable to 0.06 and 0.04 to 0.30, respectively. These results show that the quantitative evaluation by RQ-PCR is a valuable tool in the determination of dCK, dGK and 5'-NT mRNA levels in cell lines and in clinical samples which were expressed at various levels. This rapid, convenient and specific method is suitable for further studies of these genes in clinical samples. PMID- 11896544 TI - Transcription-mediated amplification and hybridisation protection assay to determine BCR-ABL transcript levels in patients with chronic myeloid leukaemia. AB - Detection of BCR-ABL transcripts in chronic myeloid leukaemia (CML) is used to confirm the diagnosis and to monitor residual disease. Quantitative techniques are required to predict response to therapy or early relapse. We have evaluated an assay in which transcription-mediated amplification (TMA) of BCR-ABL and ABL transcripts is achieved using reverse transcriptase and RNA polymerase. The products are quantified in the hybridisation protection assay (HPA) using acridinium ester-labelled DNA probes and chemiluminescence. The method is a single tube procedure which uses small amounts of RNA (<500 ng/triplicate analysis), is technically simple (requiring just two waterbaths and a luminometer), rapid (total assay time <4 h) and sensitive (capable of detecting one BCR-ABL-positive K562 cell in the presence of 10(4)-10(5) BCR-ABL-negative cells). BCR-ABL signals from patient RNA samples were quantified relative to known amounts of K562 RNA and normalised to levels of ABL. BCR-ABL/ABL ratios ranged from 0.15 to 1.59 (median 0.65) in RNA from diagnostic blood or bone marrow of 18 CML patients and were < or =0.0001 in 20 normal controls. Sequential samples analysed from six CML patients post-allogeneic bone marrow transplantation who relapsed and received donor lymphocyte infusions showed BCR ABL/ABL ratios which reflected patient status or treatment. A BCR-ABL/ABL ratio of 0.01 served as a useful arbitrary indicator value, with results above and below this value generally correlating with relapse or remission, respectively. PMID- 11896548 TI - Haploidentical 'megadose' CD34+ cell transplants for patients with acute leukemia. PMID- 11896550 TI - Second European Workshop on haploidentical stem cell transplantation (12-14 October 2000 - Perugia, Italy). PMID- 11896551 TI - Workshop on haploidentical stem cell transplantation: Chicago, Illinois, USA, 18 19 November 2000. PMID- 11896552 TI - Haploidentical 'megadose' stem cell transplantation in acute leukemia: recommendations for a protocol agreed upon at the Perugia and Chicago meetings. PMID- 11896555 TI - Application of physiological genomics to the microcirculation. AB - Physiological genomics represents a new challenge in the biological sciences-the quest to define the functions of thousands of genes that will emerge from the sequencing of the human genome and the genomes of other model organisms. Because the attention of the scientific community has focused on this task, new tools that will allow high-efficiency identification of gene function are being developed at remarkable speed. Physiological genomic approaches to understanding integrated systems function are now becoming widely used in many areas of biological research. The availability of genomic information across species has now revealed a striking degree of conservation of both gene order and function, allowing researchers to easily move from model organisms to man in the hunt for gene function. Physiological genomics approaches in the cardiovascular system have focused on disease-based models and the behavior of large vessels. In the microcirculation, genomic studies have largely been confined to the use of single gene knockouts or to the study of angiogenesis. This review summarizes the strategies for physiological genomics that are appropriate to the study of the microcirculation and discusses several key discoveries that have been made by using these approaches. PMID- 11896556 TI - DNA microarrays: their use and misuse. AB - DNA microarray represents one of the major advances in functional genomics. Its ability to study expression of several thousands of genes or even all genes in the entire genome in a single experiment has changed the way in which we address basic biomedical questions. Numerous publications have shown its utility in drug discovery, disease diagnosis, novel gene identification, and understanding complex biological systems. However, there are substantive technical issues associated with the use of this technology that limit the interpretation of microarray data. In this review, we first give an overview of DNA microarray technology and then focus on uncertainty areas of microarray technology that include making microarrays, isolation of RNA and labeling, hybridization and scanning, and data analysis. The center theme of this review is to improve microarray reproducibility by addressing common technical problems. Finally, we briefly summarize microarray's applications in biomedical research. PMID- 11896557 TI - Virally mediated gene transfer to the vasculature. AB - Gene transfer technology provides valuable tools for the study of vascular biology. By using gene transfer, effects of specific gene products can be evaluated in a highly selective manner. In recent years, techniques used for gene transfer have been adapted for applications to blood vessels, including microvessels, both in vitro and in vivo. The purpose of this review is to provide a survey of published work in this field of investigation and to discuss advantages and limitations of current methods used for gene transfer to the vasculature. PMID- 11896559 TI - Use of antisense oligonucleotides: advantages, controls, and cardiovascular tissue. AB - Antisense oligonucleotides are short pieces of synthetic, chemically modified DNA or RNA that are designed to interact by Watson-Crick base pairing with mRNA encoding a targeted protein. During the past 20 years the technology associated with the development of antisense has improved dramatically, and emerging chemistries have made antisense oligonucleotides into powerful and versatile tools to study the function of proteins in living cells. The dramatic increase in novel genomic sequence information that has recently become available has generated enormous opportunities for the development of antisense oligonucleotides capable of altering the expression level of virtually any gene. With this will come a nearly equal opportunity to determine the role of individual proteins in a vast array of cardiovascular disease. The great specificity that these compounds exhibit in vitro suggests that they may also have an exciting future for development into therapeutics useful for the treatment of human disease. This review highlights some of the advances made in the field of antisense research, placing an emphasis on uses and proper controls. PMID- 11896558 TI - Nonviral gene transfer strategies for the vasculature. AB - Major attention has been focused on the development of gene therapy approaches for the treatment of vascular diseases. In this review, we focus on an alternative use of gene therapy: the use of genetic means to study vascular cell biology and physiology. Both viral and nonviral gene transfer strategies have limitations, but because of the overwhelming inflammatory responses associated with the use of viral vectors, nonviral gene transfer methods are likely to be used more abundantly for future applications in the vasculature. Researchers have made great strides in the advancement of gene delivery to the vasculature in vivo. However, the efficiency of gene transfer seen with most nonviral approaches has been exceedingly low. We discuss how to circumvent and take advantage of a number of the barriers that limit efficient gene delivery to the vasculature to achieve high-level gene expression in appropriate cell types within the vessel wall. With such levels of expression, gene transfer offers the ability to alter pathways at the molecular level by genetically modulating the activity of a gene product, thus obviating the need to rely on pharmacological agents and their foreseen and unforeseen side effects. This genetic ability to alter distinct gene products within a signaling or biosynthetic pathway or to alter structural interactions within and between cells is extremely useful and technologically possible today. Hopefully, with the availability of these tools, new advances in cardiovascular physiology will emerge. PMID- 11896563 TI - National Cancer Institute workshop on chemopreventive properties of nonsteroidal anti-inflammatory drugs: role of COX-dependent and -independent mechanisms. AB - A workshop, "Chemopreventive properties of nonsteroidal anti-inflammatory drugs (NSAIDs): Role of COX-dependent and -independent mechanisms," sponsored by the Chemical and Physical Carcinogenesis Branch, Division of Cancer Biology of the National Cancer Institute, was held in Rockville, Maryland, on January 8, 2001. The workshop was composed of two parts: oral presentations by a series of speakers, and a group discussion of preselected topics. PMID- 11896564 TI - In utero exposure to low doses of bisphenol A lead to long-term deleterious effects in the vagina. AB - The origins of the "endocrine disrupter hypothesis" may be traced to reports on adolescent daughters born to women who had taken the highly potent synthetic estrogen, diethylstilbestrol, while pregnant, and who developed a rare form of vaginal cancer and adenocarcinoma. Bisphenol A (BPA) is an estrogenic chemical that is highly employed in the manufacture of a wide range of consumer products. Some observational studies have suggested that the amounts of BPA to which we are exposed could alter the reproductive organs of developing rodents. We examined the influence of BPA at low doses to address the questions of (a) whether in utero exposure affects the vagina of the offspring and (b) which mechanisms cause the toxic effects. Gravid Sprague-Dawley dams were administered either 0.1 (low dose) or 50 mg/kg per day BPA, the no observed effect level, or 0.2 mg/kg per day 17 alpha-ethinyl estradiol by gavage. Striking morphological changes were observed in the vagina of postpubertal offspring leading us to examine vaginal estrogen receptor (ER) expression because BPA binds to the ER alpha, which is important for growth of the vaginal epithelium. We show that the full-length ER alpha is not expressed during estrus in the vagina of female offspring exposed to either dose of BPA when compared to the control group, whereas ER alpha expression does not differ from the control group during the diestrus stage. ER alpha downregulation seems to be responsible for the observed altered vaginal morphology. PMID- 11896565 TI - High telomerase activity correlates with the stabilities of genome and DNA ploidy in renal cell carcinoma. AB - Malignant tumors have telomerase activity, which is thought to play a critical role in tumor growth. However, the relation between telomerase activity and genomic DNA status in tumor cells is poorly understood. In the present study, we examined telomerase activity in 13 clear cell type renal cell carcinomas (CRCCs) with similar clinicopathologic features by telomeric repeat amplification protocol assay (TRAP). Based on TRAP assay results, we divided the CRCCs into two groups: a high telomerase activity group and a low/no telomerase activity group. We then analyzed genomic aberration, DNA ploidy, and telomere status in these two groups by comparative genomic hybridization (CGH), laser scanning cytometry (LSC), and telomere-specific fluorescence in situ hybridization (T-FISH), respectively. CGH showed the high telomerase activity group to have fewer genomic changes than the low/no telomerase activity group, which had many genomic aberrations. Moreover, with LSC, DNA diploid cells were found more frequently in the high telomerase activity group than in the low/no telomerase activity group. In addition, T-FISH revealed strong telomere signal intensity in the high telomerase activity group compared with that of the low/no telomerase activity group. These results suggest that telomerase activity is linked to genomic DNA status and that high telomerase activity is associated with genomic stability, DNA ploidy, and telomere length in CRCC. PMID- 11896566 TI - Development of PIN and prostate adenocarcinoma cell lines: a model system for multistage tumor progression. AB - Existing prostate cancer cell lines have been derived from late stages of human prostate cancer. In this paper, we present two cell lines generated from prostatic intraepithelial neoplasia (PIN), the precursor lesion for prostate adenocarcinoma. Pr-111 and Pr-117 were established from PIN lesions that developed in the C3(1)/Tag transgenic model of prostate cancer. Pr-111 and Pr-117 cells express simian virus 40 large T antigen (SV40 Tag) and are immortalized in culture, distinguishing them from normal prostate cells. The growth rates of these two cell lines are quite different; with Pr-111 cells growing much more slowly (doubling time approximately 40 hours) compared to Pr-117 cells (doubling time approximately 22 hours), and also show significantly different growth rates in different media. Both prostate cell lines express cytokeratin and androgen receptor (AR) with Pr-111 cells demonstrating androgen-dependent growth and Pr 117 cells exhibiting androgen-responsive growth characteristics. Athymic nude mice injected with Pr-111 cells either do not develop tumors or develop tumors after a long latency period of 14 weeks. Pr-117 cells, however, develop tumors by 3 to 6 weeks, suggesting that Pr-117 cells represent a later stage of tumor progression. These two novel cell lines will be useful for studying early stages of prostate tumor development and androgen responsiveness. PMID- 11896567 TI - Transcriptional analyses of Barrett's metaplasia and normal upper GI mucosae. AB - Over the last two decades, the incidence of esophageal adenocarcinoma (EA) has increased dramatically in the US and Western Europe. It has been shown that EAs evolve from premalignant Barrett's esophagus (BE) tissue by a process of clonal expansion and evolution. However, the molecular phenotype of the premalignant metaplasia, and its relationship to those of the normal upper gastrointestinal (GI) mucosae, including gastric, duodenal, and squamous epithelium of the esophagus, has not been systematically characterized. Therefore, we used oligonucleotide-based microarrays to characterize gene expression profiles in each of these tissues. The similarity of BE to each of the normal tissues was compared using a series of computational approaches. Our analyses included esophageal squamous epithelium, which is present at the same anatomic site and exposed to similar conditions as Barrett's epithelium, duodenum that shares morphologic similarity to Barrett's epithelium, and adjacent gastric epithelium. There was a clear distinction among the expression profiles of gastric, duodenal, and squamous epithelium whereas the BE profiles showed considerable overlap with normal tissues. Furthermore, we identified clusters of genes that are specific to each of the tissues, to the Barrett's metaplastic epithelia, and a cluster of genes that was distinct between squamous and non-squamous epithelia. PMID- 11896568 TI - Expression of Bcl-2 family member Bid in normal and malignant tissues. AB - Bid is the only known Bcl-2 family member that can function as an agonist of proapoptotic Bcl-2-related proteins such as Bax and Bak. Expression of the proapoptotic Bcl-2 family protein Bid was assessed by immunoblotting and immunohistochemical methods in normal murine and human tissues, and in several types of human cancers and tumor cell lines. Bid expression in normal tissues varied widely, with prominent Bid immunostaining occurring in several types of short-lived cells (e.g., germinal center B cells, peripheral blood granulocytes, differentiated keratinocytes) and in apoptosis-sensitive cells (e.g., adult neurons). Analysis of Bid expression by immunostaining of 100 colon, 95 ovarian, and 254 prostate cancers, as well as 59 brain tumors and 50 lymphomas, revealed evidence of altered Bid regulation in some types of cancers. Correlations with clinical outcome data revealed association of higher levels of Bid with longer recurrence-free survival in men with locally advanced (T3 stage) prostate cancer (P=0.04). Immunoblot analysis of Bid protein levels in the NCI's panel of 60 human tumor cell lines revealed a correlation between higher levels of Bid and sensitivity to ribonucleotide reductase (RR)-inhibiting drugs (P<0.0005). Overexpression of Bid in a model tumor cell line by gene transfection resulted in increased sensitivity to apoptosis induction by a RR inhibitor. Taken together, these observations suggest a potential role for Bid in tumor responses to specific chemotherapeutic drugs, and lay a foundation for future investigations of this member of the Bcl-2 family in healthy and diseased tissues. PMID- 11896569 TI - Differential expression of critical cellular genes in human lung adenocarcinomas and squamous cell carcinomas in comparison to normal lung tissues. AB - The Atlas human cDNA expression array was used to evaluate gene expression profile changes in the genesis of human lung adenocarcinomas and squamous cell carcinomas. Gene expression changes between adenocarcinomas and squamous cell carcinomas were also analyzed. Of the 588 gene targets, 262 genes were expressed in these tissues and, of these, 45 genes were differentially expressed by at least two-fold in tumor tissues compared to corresponding normal tissues. Semiquantitative reverse-transcriptase polymerase chain reaction was used to confirm gene expression changes. Only those genes that reflected changes in >50% of the analyzed tissues were included in the final analysis. Ultimately, 26 genes were evaluated with 14 genes overexpressed and 12 genes underexpressed compared to matching normal lung tissues. Although similar expression changes were detected in adenocarcinomas and squamous cell carcinomas for most of the genes analyzed, some subtype-specific differences were also found. Genes encoding cell cycle regulators, intracellular signal transducers, cell receptor and adhesion molecules, growth factors, oncogenes, and apoptotic effectors were differentially expressed in this study. These gene expression changes may directly contribute to the initiation or progression of human lung cancer or may be secondary effects of the tumorigenesis process. Regardless, many of these differences may be useful in the diagnosis and/or treatment of this deadly disease. PMID- 11896570 TI - Deregulated expression of the human tumor marker CEA and CEA family member CEACAM6 disrupts tissue architecture and blocks colonocyte differentiation. AB - Human carcinoembryonic antigen (CEA) and the CEA family member CEACAM6 (formerly nonspecific cross-reacting antigen [NCA]) function in vitro, at least, as homotypic intercellular adhesion molecules and, in model systems, can block the terminal differentiation and anoikis of several different cell types. We have recently demonstrated that the increased cell surface levels of CEA and CEACAM6 in purified human colonocytes from freshly excised, well to poorly differentiated colon carcinomas are inversely correlated with the degree of cellular differentiation. Thus, deregulated expression of CEA/CEACAM6 could directly contribute to colon tumorigenesis by the inhibition of terminal differentiation and anoikis. Evidence against this view includes the common observation of increased CEA/CEACAM6 expression as normal colonocytes differentiate in their migration up colonic crypt walls. We report here the direct effects of deregulated overexpression of CEA/CEACAM6, at levels observed in colorectal carcinomas, on the differentiation of two human colonic cell lines, SW-1222 and Caco-2. Stable transfectants of both of these cell lines that constitutively express 10- to 30-fold higher cell surface levels of CEA/CEACAM6 than endogenous levels failed to polarize and differentiate into glandular structures in monolayer or 3D culture or to form colonic crypts in a tissue architecture assay in nude mice. In addition, these transfectants were found to exhibit increased tumorigenicity in nude mice. These results thus support the contention that deregulated overexpression of CEA and CEACAM6 could provide a tumorigenic contribution to colon carcinogenesis. PMID- 11896571 TI - Radiotherapy and antiangiogenic TM in lung cancer. AB - Tetrathiomolybdate (TM) is a potent nontoxic orally delivered copper complexing agent under development for the last several years for the treatment of Wilson's disease. It has been shown to block angiogenesis in primary and metastatic tumors. Therefore, the combination of cytotoxic radiotherapy (RT) and antiangiogenic TM could target both the existing tumor and the tumor microvasculature in a comprehensive strategy. Using a Lewis lung high metastatic (LLHM) carcinoma mouse tumor model, we demonstrate that the combination of TM and RT is more effective than either used as monotherapy. We also show that their therapeutic effects are additive, with no additional toxicity. We show that TM has no significant cytotoxicity in vitro against LLHM tumor cells, further supporting the antiangiogenic mechanism for its action. PMID- 11896574 TI - A positive feedback mechanism in the transcriptional activation of Apaf-1 by p53 and the coactivator Zac-1. AB - p53 exerts its tumor suppressor effects by activating genes involved in cell growth arrest and programmed cell death. The p53 target genes inducing growth arrest are well defined whereas those inducing apoptosis are not fully characterized. Proapoptotic activity of p53 was shown to involve several genes like Bax, Noxa and Puma, which may function in the release of cytochrome c from the mitochondria. Cytochrome c associates with Apaf-1 and caspase 9 to form the apoptosome. Genetic and cellular data indicate that Apaf-1 deficiency abrogates the apoptotic effect of p53 and substitutes for p53 loss in promoting tumor formation. Here we show that Apaf-1, the mammalian homologue of C. elegans CED4, is a direct target of p53 as demonstrated by gel shift analysis of the target site sequence in the presence of p53 and by Apaf-1 promoter-luciferase assays. We also show that the p53 activation of the Apaf-1 luciferase construct can be enhanced by the putative tumor suppressor gene product, Zac-1, a transcription factor that has previously been shown to inhibit cell proliferation. Furthermore, we demonstrate that Zac-1 is a possible direct target of p53 since the sequence upstream to the first coding exon of Zac-1 contains a p53 recognition site and the luciferase construct containing this region is activated by p53. These results suggests the existence of a tightly controlled self amplifying mechanism of transcriptional activation leading to apoptosis by p53. PMID- 11896575 TI - The SH2-containing inositol 5-phosphatase (SHIP)-1 is implicated in the control of cell-cell junction and induces dissociation and dispersion of MDCK cells. AB - Hepatocyte growth factor (HGF) induces the breakdown of cell junction and the dispersion of colonies of epithelial cells, providing a model system for the investigation of the molecular mechanisms of one of the important aspects of tumorogenesis. We have previously reported that the SH2-domain-containing inositol 5'phosphatase (SHIP)-1 binds to c-Met, and potentiated HGF-mediated branching tubulogenesis. In this study, we describe the establishment of MDCK cell lines which express MycHis-tagged SHIP-1 at different levels. Expression of SHIP-1 in MDCK cells at a high level resulted in cell morphology characteristic of an epithelial-mesenchymal like transition; cells lost cortical actin, developed actin stress fibers and gained spontaneous motility without treatment of HGF. When the level of MycHis-tagged SHIP-expression was relatively low, transfectants partially lost cortical actin and phalloidin stained puncta appeared at cell-cell junctions even in the absence of HGF. The treatment of MAP kinase inhibitor, PD98059, did not influence SHIP-1 mediated alteration of adherens-junction of MDCK cells, while, phosphatidylinositol 3 (PI 3)- kinase inhibitor, LY294002, drastically reduced SHIP-1 mediated phenotype. Furthermore, expression of a mutant SHIP-1 lacking catalytic activity in MDCK cells did not alter the cortical actin distribution and HGF-mediated MAP and Akt kinase phosphorylation, but suppressed HGF induced cell dispersion, suggesting that phosphatase activity is important for cytoskeleton rearrangement and cell dispersion. PMID- 11896572 TI - Radiation-induced phosphorylation of Chk1 at S345 is associated with p53 dependent cell cycle arrest pathways. AB - Because DNA damage-inducible cell cycle checkpoints are thought to protect cells from the lethal effects of ionizing radiation, a better understanding of the mechanistic functions of cell cycle regulatory proteins may reveal new molecular targets for cancer therapy. The two major regulatory proteins of G2 arrest are Chk1 and p53. Yet, it is unclear how these two proteins interact and coordinate their functional roles during radiation-induced G2 arrest. To determine Chk1's role in p53-dependent G2 arrest, we used p53 proficient cells and examined expression of G2 arrest proteins under conditions in which G2 arrest was inhibited by the staurosporine analog, UCN-01. We found that UCN-01 inhibited both G1 and G2 arrest in irradiated p53 proficient cells. The arrest inhibition was associated with suppression of radiation-induced expression of both p21 and 14-3-3 sigma -- two known p53-dependent G2 arrest proteins. The suppression occurred despite normal induction of p53 and normal phosphorylation of p53 at S20 and Cdc25C at S216 -- the two known substrates of Chk1 kinase activity. In contrast, we showed that radiation-induced phosphorylation of Chk1 at S345 was associated with binding of Chk1 to p53, p21, and 14-3-3 sigma, and that UCN-01 inhibited S345 phosphorylation. We suggest that DNA damage-induced phosphorylation of Chk1 at S345, and subsequent p53 binding, links Chk1 with p53 downstream responses and may provide a coordinated interaction between DNA damage responses and cell cycle arrest functions. PMID- 11896576 TI - Sp1 as G1 cell cycle phase specific transcription factor in epithelial cells. AB - Sp1 binding sites have been identified in enhancer/promoter regions of several growth and cell cycle regulated genes, and it has been shown that Sp1 is increasingly phosphorylated in G1 phase of the cell cycle. Interactions of Sp1 with proteins involved in control of cell cycle and tumor formation have been reported. Here we show that expression of Sp1 protein predominates in the G1 phase of the cell cycle in epithelial cells. This is achieved by proteasome dependent degradation. Inhibition of endogeneous Sp1 activity by a dominant negative Sp1 mutant was associated with a cell cycle arrest in G1 phase, a strongly reduced expression of cyclin D1, the EGF-receptor and increased levels of p27Kip1. We have thus identified Sp1 as an important regulator of the cell cycle in G1 phase. PMID- 11896577 TI - Centrosome overduplication, increased ploidy and transformation in cells expressing endoplasmic reticulum-associated cyclin A2. AB - Cyclin A2 is predominantly, but not exclusively, localized in the nucleus from G1/S transition onwards. It is degraded when cells enter mitosis after nuclear envelope breakdown. We previously showed that a fusion protein (S2A) between the hepatitis B virus (HBV) surface antigen protein and a non-degradable fragment of human cyclin A2 (Delta152) resides in the endoplasmic reticulum membranes, escapes degradation and transforms normal rat fibroblasts. The present study investigates whether cytoplasmic cyclin A2 may play a role in oncogenesis. We show that the sequestration of non-degradable cyclin A2-Delta152 by a cellular ER targeting domain (PRL-A2) leads to cell transformation when coexpressed with activated Ha-ras. REF52 cells constitutively expressing PRL-A2 are found to have a high incidence of multinucleate giant cells, polyploidy and abnormal centrosome numbers, giving rise to the nucleation of multipolar spindles. Injection of these cells into athymic nude mice causes tumors, even in the absence of a cooperating Ha-ras oncogene. These results demonstrate that, independently of any viral context, an intracellular redistribution of non-degradable cyclin A2 is capable of deregulating the normal cell cycle to the point where it promotes aneuploidy and cancer. PMID- 11896578 TI - Mouse homologue of HOS (mHOS) is overexpressed in skin tumors and implicated in constitutive activation of NF-kappaB. AB - NF-kappaB transcription factor is activated upon ubiquitination and subsequent proteolysis of its inhibitor IkappaB. The phosphorylation-dependent ubiquitination is mediated by SCF E3 ubiquitin ligase. In this study, we identified a novel murine F-box/WD40 repeat-containing protein, mHOS (a homologue of HOS/betaTrCP2). mHOS efficiently binds Skp1 protein (a 'core' component of SCF ubiquitin ligase), and phosphorylated IkappaB(alpha). We found that mHOS associates with SCF-ROC1 E3 ubiquitin ligase activity. We have also observed that mHOS is overexpressed in chemically-induced mouse skin tumors, and its overexpression (but not accelerated IkappaB phosphorylation) coincides with the accelerated degradation of IkappaB in vivo. The role of mHOS in the constitutive activation of NF-kappaB in skin carcinogenesis is discussed. PMID- 11896579 TI - Involvement of intact HPV16 E6/E7 gene expression in head and neck cancers with unaltered p53 status and perturbed pRb cell cycle control. AB - We have identified parameters which define a causal role of HPV16 in head and neck cancer. Twenty-eight tumours which were typed positive for HPV16 DNA, were comprehensively analysed for expression of the viral oncogenes E6 and E7, the status of the p53 gene, and the protein status of pRb and p16(INK4a). In a subset of cases, we have searched for integrated viral DNA, and have determined the genomic status of the E6 gene. Expression of E6/E7 was found in 12 tumours most of which were derived from the oropharynx, whereas p53 mutations were present in 13 tumours from various sites. The tumours either carried p53 mutations but did not express E6/E7, or they did express E6/E7 but were p53-wild-type. Coexistence of E6/E7 expression with a mutated p53 was found in only one case. Strikingly, in most p53-mutated tumours without E6/E7 expression, we found the E6 gene to be disrupted. E6/E7 expression was associated with reduced pRb and overexpressed p16(INK4a). Viral-cellular fusion transcripts were found in two cases. Our data demonstrate that HPV16 DNA-positivity in head and neck cancers is not indicative of a causal role. A causal role of HPV16 in head and neck cancer is defined by: E6/E7 expression, viral integration with an intact E6 gene, and perturbation of pRb cell cycle control. Mostly, the p53 gene is wild-type. PMID- 11896580 TI - Recurrent allelic deletions at mouse chromosomes 4 and 14 in Myc-induced liver tumors. AB - Transgenic mice expressing the c-Myc oncogene driven by woodchuck hepatitis virus (WHV) regulatory sequences develop hepatocellular carcinoma with a high frequency. To investigate genetic lesions that cooperate with Myc in liver carcinogenesis, we conducted a genome-wide scan for loss of heterozygosity (LOH) and mutational analysis of beta-catenin in 37 hepatocellular adenomas and carcinomas from C57BL/6 x castaneus F1 transgenic mice. In a subset of these tumors, chromosome imbalances were examined by comparative genomic hybridization (CGH). Allelotyping with 99 microsatellite markers spanning all autosomes revealed allelic imbalances at one or more chromosomes in 83.8% of cases. The overall fractional allelic loss was rather low, with a mean index of 0.066. However, significant LOH rates involved chromosomes 4 (21.6% of tumors), 14, 9 and 1 (11 to 16%). Interstitial LOH on chromosome 4 was mapped at band C4-C7 that contains the INK4a/ARF and INK4b loci, and on chromosome 14 at band B-D including the RB locus. In man, the homologous chromosomal regions 9p21, 13q14 and 8p21-23 are frequently deleted in liver cancer. LOH at chromosomes 1 and 14, and beta catenin mutations (12.5% of cases) were seen only in HCCs. All tumors examined were found to be aneuploid. CGH analysis of 10 representative cases revealed recurrent gains at chromosomes 16 and 19, but losses or deletions involving mostly chromosomes 4 and 14 generally prevailed over gains. Thus, Myc activation in the liver might select for inactivation of tumor suppressor genes on regions of chromosomes 4 and 14 in a context of low genomic instability. Myc transgenic mice provide a useful model for better defining crosstalks between oncogene and tumor suppressor pathways in liver tumorigenesis. PMID- 11896581 TI - Epidermal Ras blockade demonstrates spatially localized Ras promotion of proliferation and inhibition of differentiation. AB - While important in carcinogenesis, the role of Ras in normal self-renewing tissues such as epidermis is unclear. To address this, we altered Ras function in undifferentiated and differentiating epidermal layers. Ras blockade within undifferentiated basal epidermal cells leads to decreased integrin expression, diminished growth capacity and induction of differentiation. Ras blockade in post mitotic suprabasal epidermis exerts no effect. In contrast, regulated Ras and Raf activation inhibits differentiation. These findings indicate that spatially restricted Ras/Raf signaling divides epidermis into an undifferentiated proliferative compartment and a differentiating post-mitotic compartment and suggest a new role for Ras in tissue homeostasis. PMID- 11896583 TI - Bcl-2 expression delays hepatocyte cell cycle progression during liver regeneration. AB - Bcl-2 is the prototype of a family of genes that prevent apoptosis. However, several reports indicate that Bcl-2 may also act as a cell cycle modulator. In several human tumors, Bcl-2 expression correlates with a more favorable prognosis and lower tumor proliferative activity. We have shown that Bcl-2 expression delays liver tumor development in transgenic mice even when the gene is turned on shortly before the time of tumor development. We hypothesized that Bcl-2 may delay liver tumorigenesis by interfering with hepatocyte proliferation. To test whether Bcl-2 expression may act on hepatocyte replication we studied liver regeneration in Bcl-2 transgenic mice and wild-type littermates. DNA replication was delayed by approximately 8 h in Bcl-2 transgenic mice compared to the timing of the response in wild-type littermates. Cyclin D expression showed no alterations in the regenerating liver of Bcl-2 transgenic mice. In contrast, there was a delay in the expression of p107, cyclin E and in the activity of cyclin E/cdk 2 activity. These results show that Bcl-2 expression delays cell cycle progression in hepatocytes and suggests that it acts at a step involving cyclin E and p107. PMID- 11896582 TI - Retinoic acid inhibits hepatic Jun N-terminal kinase-dependent signaling pathway in ethanol-fed rats. AB - Retinoic acid (RA) supplementation suppresses ethanol-enhanced hepatocyte hyperproliferation in rats; however, little is known about the mechanism(s). Here, we investigated whether RA affects the protein kinase signaling pathways in the liver tissues of rats fed with a high dose of ethanol for a prolonged period of time (6 months). Results show that there were greater levels of phosphorylated Jun N-terminal kinase (JNK) and phosphorylated c-Jun protein, but not total JNK protein, in livers of ethanol-fed rats vs those of controls. Moreover, ethanol feeding to rats increased the levels of phosphorylated mitogen-activated protein kinase kinase-4 (MKK-4) and decreased the levels of mitogen-activated kinase phosphatase-1 (MKP-1) in liver tissue. However, hepatic levels of phosphorylated p38 protein and total-p38 protein were not altered by the ethanol treatment. In contrast, all-trans-RA supplementation at two doses in ethanol-fed rats greatly attenuated the ethanol-induced hepatic phosphorylation of MKK-4, phosphorylated JNK and c-Jun proteins. The level of MKP-1 was increased in ethanol-fed rats supplemented with all-trans-RA. Further, ethanol-induced hepatocyte hyperproliferation, measured by immunostaining for proliferating cell nuclear antigen, were markedly decreased by all-trans-RA supplementation. Interestingly, hepatic apoptosis in the liver of ethanol-fed rats after 6 months of treatment decreased significantly. This decrease of hepatic apoptosis in ethanol-fed rats was prevented by all-trans-RA supplementation in a dose-dependent manner. The results from these studies indicate that restoration of RA homeostasis is critical for the regulation of JNK-dependent signaling pathway and apoptosis in the liver of ethanol-fed rats. PMID- 11896584 TI - Phosphorylation of forkhead transcription factors by erythropoietin and stem cell factor prevents acetylation and their interaction with coactivator p300 in erythroid progenitor cells. AB - The mammalian forkhead transcription factors, FOXO3a (FKHRL1), FOXO1a (FKHR) and FOXO4 (AFX) are negatively regulated by PKB/Akt kinase. In the present study we examined the engagement of forkhead family of transcription factors in erythropoietin (Epo)- and stem cell factor (SCF)-mediated signal transduction. Our data show that all three forkhead family members, FOXO3a, FOXO1a and FOXO4 are phosphorylated in human primary erythroid progenitors. Experiments performed to determine various upstream signaling pathways contributing to phosphorylation of forkhead family members show that only PI-3-kinase pathway is required for inactivation of FOXO3a. Our data also demonstrate that during Epo deprivation FOXO3a interacts with the transcriptional coactivator p300 and such interaction is disrupted by stimulation of cells with Epo. To determine the domains in FOXO3a, mediating its interaction with p300, we performed GST pull-down assays and found that the N-terminus region containing the first 52 amino acids was sufficient for binding p300. Finally, our data demonstrate that FOXO3a and FOXO1a are acetylated during growth factor deprivation and such acetylation is reversed by stimulation with Epo. Thus mammalian forkhead transcription factors are involved in Epo and SCF signaling in primary erythroid progenitors and may play a role in the induction of apoptotic and mitogenic signals. PMID- 11896585 TI - Differences in DNA binding properties between E2F1 and E2F4 specify repression of the Mcl-1 promoter. AB - E2F1 is a potent inducer of apoptosis whereas its relative, E2F4, generally does not promote cell death. Other work from our laboratory has demonstrated that E2F1 can directly bind and represss the Mcl-1 promoter - contributing to E2F1-mediated apoptosis. Here we show that while E2F1 can repress the Mcl-1 promoter, other members of the E2F family (such as E2F4) cannot. Characterization of the Mcl-1 promoter demonstrates that the -143/+10 region is critical for E2F1-mediated downregulation. We demonstrate that the ability of E2F1 to repress the Mcl-1 promoter correlates with its ability to bind within the required -143/+10 region of this promoter. In contrast, E2F4 is unable to bind to the -143/+10 region of the Mcl-1 promoter. We propose that E2F4 is unable to repress the Mcl-1 promoter primarily as a result of insufficient binding to the essential regulatory region. This is the first evidence of DNA binding specificity among E2F family members that results in differential regulation of a naturally occurring promoter. PMID- 11896586 TI - High levels of the onco-protein Gfi-1 accelerate T-cell proliferation and inhibit activation induced T-cell death in Jurkat T-cells. AB - Gfi-1 is a nuclear zinc finger protein with the activity of a transcriptional repressor and the ability to predispose for the development of T-cell lymphoma when expressed constitutively at high levels. Whereas thymic T-cell precursors express endogenous Gfi-1, mature peripheral T-cells lack Gfi-1 but upregulate its expression transiently after antigenic stimulation and activation of Erk1/2 demonstrating a role of Gfi-1 in T-cell activation. Here we show that constitutive expression of Gfi-1 accelerates S phase entry of primary, resting T cells upon antigenic stimulation. In addition, high level Gfi-1 expression inhibits phorbol ester induced G1 arrest and activation induced cell death in Jurkat T-cells. We demonstrate that these effects of Gfi-1 concur with lower absolute levels and hyperphosphorylation of the pocket protein pRb. Moreover, phorbol ester induced expression of the negative cell cycle regulator p21(WAF1) is blocked in the presence of Gfi-1. These findings suggest that Gfi-1 contributes to T-cell lymphomagenesis by overriding a late G1 cell cycle checkpoint which controls activation induced death and S phase entry of T-cells. PMID- 11896587 TI - Role of MAP kinases in UVB-induced phosphorylation of p53 at serine 20. AB - Phosphorylation of the p53 tumor suppressor protein is one of the key regulatory steps in its activation process. Serine 20 phosphorylation of p53 has been shown to be required for the activation of p53 following UV radiation, but the signaling pathway mediating UV-induced phosphorylation is unknown. Here, we determined the role of MAP kinases in UVB-induced phosphorylation and found that JNKs are directly involved in the phosphorylation of p53 at serine 20. In a mouse JB6 epidermal cell line, dominant negative JNK1 abrogated UVB-induced phosphorylation of p53 at serine 20, whereas dominant negative p38 kinase or its inhibitor, SB202190, partially attenuated the phosphorylation. In contrast, dominant negative ERK2 or the MEK1 inhibitor, PD98059, had no effect on p53 phosphorylation at serine 20. Importantly, UVB-activated or active recombinant JNK1/2, or the p38 kinase downstream target, MAPKAPK-2, but not ERKs or p38 kinase, phosphorylated p53 at serine 20 in vitro. Furthermore, phosphorylation of p53 at serine 20 by UVB-activated JNKs and UVB-induced p53-dependent transcriptional activity were suppressed in Jnk1 or Jnk2 knockout (Jnk1(-/-) or Jnk2(-/-)) cells. Additionally, Jnk1(-/-), Jnk2(-/-), and p53-deficient (p53(-/ )) cells, as well as re-introduction of a p53 mutant with substitution of serine 20 to alanine into p53(-/-) cells, were defective for UVB-induced apoptosis. These findings strongly suggest that JNKs are the major direct signaling mediators of UVB-induced p53 phosphorylation at serine 20. PMID- 11896588 TI - Rho regulates p21(CIP1), cyclin D1, and checkpoint control in mammary epithelial cells. AB - The small GTPase Rho is important for cell cycle progression and Ras transformation in fibroblasts. However, it is unclear whether Rho is needed for proliferation in other cell types, and its targets in promoting normal cell cycle progression are unknown. Here, we demonstrate that Rho is required for G1 to S progression in MCF10A mammary epithelial cells, both in response to EGF and in response to oncogenic Ras. We describe two effects of Rho, the repression of p21(CIP1) and the induction of cyclin D1, that may underlie its role in promoting S phase entry. The Rho inhibitor, C3 exotransferase, induced p21(CIP1) both in EGF-stimulated and V12Ras-expressing cells. In addition, C3 blocked EGF stimulated cyclin D1 promoter activity whereas V14RhoA induced the cyclin D1 promoter and cooperated with V12Ras in cyclin D1 induction. Finally, a high proportion of cells co-expressing V14RhoA and V12Ras displayed lobulated, polyploid nuclei that were actively synthesizing DNA. Our results demonstrate that Rho plays a fundamental role in promoting Ras-dependent S phase entry in mammary epithelial cells, whether in response to normal or oncogenic signaling, and indicate that in cells expressing oncogenic Ras, the activation of Rho diminishes p21(CIP1) expression, increases cyclin D1 promoter activity, and uncouples DNA synthesis from mitosis. PMID- 11896589 TI - Deregulated c-Myc prematurely recruits both Type I and II CD95/Fas apoptotic pathways associated with terminal myeloid differentiation. AB - Previously we have reported that deregulated expression of c-myc in normal and leukemic myeloid cells blocked differentiation and, concomitantly, induced p53 independent apoptosis. Here, we show that this morbidity was due to premature recruitment of the Fas/CD95 cell death pathway which normally operates to induce apoptosis at the end of the terminal myeloid differentiation program. Analysis of the regulated components of this pathway revealed that IL6-mediated induction of differentiation resulted in rapid cell surface expression of CD95 receptor. Deregulated c-myc prevented the downregulation of CD95 ligand by maintaining its transcription, but caused premature downregulation of c-FLIP. First, the Type II (mitochondria-dependent, bcl-2-sensitive) and, then, the Type I (mitochondria independent, bcl-2-insensitive) pathway were activated. Stable exogenous c-FLIP expression completely rescued the apoptotic phenotype. Furthermore, when the deregulated c-myc transgene was stably transduced into bone marrow cells from Fas(lpr/lpr) (CD95 receptor mutant) and FasL(gld/gld) (CD95 ligand mutant) mice, cell death was significantly suppressed relative to c-myc-transduced wild type bone marrow cells upon induction of differentiation. These data indicate that c myc-mediated apoptosis associated with blocks in myeloid differentiation is dependent on the Fas/CD95 pathway. Our findings offer important new insights into understanding how deregulated c-myc alters normal blood cell homeostasis, and how additional mutations might promote leukemogenesis. PMID- 11896590 TI - Mutational analysis of the transcriptional activation domains of v-Myb. AB - A minimal transcription activation domain of the v-Myb oncoprotein was initially mapped to a central cluster of charged residues using GAL4-Myb fusion proteins. This region has been proposed to interact directly with the CBP co-activator in animal cells. Regions flanking this central domain of v-Myb are required for transcriptional activation by the native, unfused protein in both mammalian cells and in budding yeast. To identify the critical residues for transcriptional activation, we have now subjected the minimal activation domain and flanking regions including the heptad leucine repeat to random PCR-mediated mutagenesis. We found that the entire region examined can endure extensive substitutions without affecting transcriptional activation by v-Myb in budding yeast. The few mutations that did affect transcriptional activation altered acidic residues within the minimal activation domain or the heptad leucine repeat region, rather than leucine residues. Remarkably, there was a strong concordance between transcriptional activation in animal cells and in budding yeast, even though budding yeast have no known homologue of CBP or related co-activators. In contrast, there was not a strong correlation between transcriptional activation and oncogenic transformation. PMID- 11896591 TI - TGF-beta1 acts as a tumor suppressor of human malignant keratinocytes independently of Smad 4 expression and ligand-induced G(1) arrest. AB - This study examined the role of TGF-beta1 in human keratinocyte malignancy. Two carcinoma-derived human oral keratinocyte cell lines, BICR 31 and H314, were selected on the basis of their known resistance to TGF-beta1-induced G(1) arrest, the presence of wild type TGF-beta cell surface receptors and normal Ras. Smad 4 protein was undetectable in both cell lines, but Smad 2 and Smad 3 were expressed at levels comparable with a fully TGF-beta responsive cell line, and treatment of the cells with TGF-beta1 resulted in the phosphorylation of Smad 2. Treatment with exogenous TGF-beta1 resulted in a failure to induce transcription from an artificial Smad-dependent promoter and a failure to down-regulate c-myc, but resulted in an up-regulation of AP-1 associated genes (Fra-1, JunB and fibronectin). Transient transfection of Smad 4 into BICR 31 restored TGF-beta1 induced growth inhibition and Smad-dependent transcriptional activation. Protracted treatment of cells with exogenous TGF-beta1 resulted in the attenuation of cell growth in vitro. To over-express TGF-beta1, both cell lines were transfected with latent TGF-beta1 cDNA; neutralization studies of conditioned media demonstrated that whilst the majority of the peptide was in the latent form, a small proportion was present as the active peptide. Cells that over-expressed endogenous TGF-beta1 grew more slowly in vitro compared to both the vector-only controls and cells that did not over-express the peptide. Orthotopic transplantation of cells that over-expressed endogenous TGF-beta1 to the floor of the mouth in athymic mice resulted in marked inhibition of primary tumor formation compared to controls. Expression of a dominant-negative TGF-beta type II receptor in cells that over-expressed endogenous TGF-beta1 resulted in enhanced cell growth in vitro and diminished the tumor suppressor effect of the ligand in vivo, indicating that the endogenous TGF-beta1 was acting in an autocrine capacity. The results demonstrate that over-expression of endogenous TGF-beta1 in human malignant oral keratinocytes leads to growth inhibition in vivo and tumor suppression in vitro by mechanisms that are independent of Smad 4 expression and TGF-beta1-induced G(1) arrest. PMID- 11896592 TI - Thioredoxin post-transcriptional regulation by H19 provides a new function to mRNA-like non-coding RNA. AB - Classically, the functional product of coding genes is a protein whose synthesis is directed by an mRNA-template. However, in the last few years several genes yielding an mRNA-like non-coding RNA as a functional product have been identified. In most cases these transcripts are synthesized by the RNA polymerase II, capped, spliced and polyadenylated, like classical mRNA. These latter have non-conserved open reading frames and seem to be untranslated. Consequently, it has been proposed and admitted that these genes act at the RNA level, and are so called 'riboregulators'. H19 belongs to this class of gene and its role remains a matter of debate: for some authors it is an oncogene, for others a tumour suppressor. Here, we demonstrate, using a proteomic approach, that an H19 overexpression in human cancerous mammary epithelial cells stably transfected with genomic DNA containing the entire H19 gene is responsible for positively regulating at the post-transcriptional level the thioredoxin, a key protein of the cellular redox metabolism. Interestingly, this protein accumulates in many cancerous tissues, such as breast carcinomas in which we have also demonstrated an overexpression of the H19 gene. PMID- 11896594 TI - PML NBs associate with the hMre11 complex and p53 at sites of irradiation induced DNA damage. AB - PML nuclear bodies (PML NBs) respond to many cellular stresses including viral infection, heat shock, arsenic and oncogenes and have been implicated in the regulation of p53-dependent replicative senescence and apoptosis. Recently, the hMre11/Rad50/NBS1 repair complex, involved in Double Strand Breaks (DSBs) repair, was found to colocalize within PML NBs, suggesting a role for these nuclear sub domains in the DNA repair signalling pathway. We report here that in normal human fibroblasts, after ionizing radiation (IR), the PML NBs are modified and recognize sites of DNA breaks (ssDNA breaks and DSBs). Eight to 12 h after radiation PML NBs associate with hMre11 Ionizing Radiation-Induced Foci (IRIF), and subsequently with p53 within discrete foci. The PML, hMre11 and p53 colocalizing structures mark sites of DSBs as identified by immunolocalization with anti phosphorylated histone gamma-H2AX. Furthermore, we demonstrate that ionizing radiation induces the stable association of p53 with hMre11 and PML. These results suggest that the PML NBs are involved in the recognition and/or processing of DNA breaks and possibly in the recruitment of proteins (p53 and hMre11) required for both checkpoint and DNA-repair responses. PMID- 11896595 TI - Tumour p53 mutations exhibit promoter selective dominance over wild type p53. AB - The tumour suppressor gene p53 is frequently mutated in human cancer. Tumour derived p53 mutants are usually transcriptionally inactive, but some mutants retain the ability to transactivate a subset of p53 target genes. In addition to simple loss of function, some p53 mutants may be carcinogenic through a dominant negative mechanism. Aiming at a more general classification of p53 mutants into predictive functional categories it is important to determine (i) which p53 mutants are dominant, (ii) what features characterize dominant mutants and (iii) whether dominance is target gene specific. The ability of 71 p53 mutants to inhibit wild type p53 was determined using a simple yeast transcriptional assay. Approximately 30% of the mutants were dominant. They preferentially affect highly conserved amino acids (P<0.005), which are frequently mutated in tumours (P<0.005), and usually located near the DNA binding surface of the protein (P<0.001). Different tumour-derived amino acid substitutions at the same codon usually have the same dominance phenotype. To determine whether the ability of p53 mutants to inhibit wild type p53 is target gene specific, the dominance towards p21, bax, and PIG3 binding sites was examined. Approximately 40% of the 45 mutants examined were dominant for the p21 (17/45) or PIG3 (20/45) responsive elements and 71% (32/45) were dominant for the bax responsive element. These differences are statistically significant (p21 vs bax, P<0.003; bax vs PIG3, P<0.02, Fisher's exact test) and defined a hierarchy of dominance. Finally, we extended the analysis to a group of mutants isolated in BRCA-associated tumours, some of which retained wild type level of transcription in yeast as well as in human cells, but show gain of function in transformation assays. Since transformation assays require transdominant inhibition of the endogenous wild type allele, one possible explanation for the behaviour of the BRCA-associated mutants is that they adopt conformations able to bind DNA alone but not in mixed tetramers with wild type p53. The yeast data do not support this explanation, because all BRCA-associated mutants that behaved as wild type in transcription assay were recessive in dominance assays. PMID- 11896596 TI - Relations between clusters of oxidatively damaged nucleotides and active or open nucleosomes in the rat Nth 1 gene. AB - The distribution of oxidative damage to bases such as 8-hydroxyguanine (8-OH Gua), was determined at the nucleotide level of resolution using the ligation mediated PCR technique. Administration of a renal carcinogen, ferric nitrilotriacetate (Fe-NTA), is known to induce oxidative stress and subsequent formation of 8-OH-Gua in the kidney. Whole genomic DNA was isolated from the rat kidney with or without Fe-NTA treatment and then digested with formamidopyrimidine-DNA glycosylase (Fpg). As a target, we focused on the gene of a DNA repair enzyme for thymine glycol, Nth 1. Cleaved signals were found in exon 1 and exon 3, but not exon 5. Nucleosomes in these regions, enriched in damaged nucleotides, were highly accessible to micrococcal nuclease, especially in the kidney. Taking into account the function of the protein segment encoded by these regions, we discussed the molecular mechanism of the restricted formation of the damaged nucleotides. PMID- 11896597 TI - Localization of the human oxytocin receptor in caveolin-1 enriched domains turns the receptor-mediated inhibition of cell growth into a proliferative response. AB - In this study, we investigated the functional role of the localization of human OTR in caveolin-1 enriched membrane domains. Biochemical fractionation of MDCK cells stably expressing the WT OTR-GFP indicated that only minor quantities of receptor are partitioned in caveolin-1 enriched domains. However, when fused to caveolin-2, the OTR protein proved to be exclusively localized in caveolin-1 enriched fractions, where it bound the agonist with increased affinity and efficiently coupled to Galpha(q/11). Interestingly, the chimeric protein was unable to undergo agonist-induced internalization and remained confined to the plasma membrane even after prolonged agonist exposure (120 min). A striking difference in receptor stimulation was observed when the OT-induced effect on cell proliferation was analysed: stimulation of the human WT OTR inhibited cell growth, whereas the chimeric protein had a proliferative effect. These data indicate that the localization of human OTR in caveolin-1 enriched microdomains radically alters its regulatory effects on cell growth; the fraction of OTR residing in caveolar structures may therefore play a crucial role in regulating cell proliferation. PMID- 11896598 TI - Activation of STAT5 triggers proliferation and contributes to anti-apoptotic signalling mediated by the oncogenic Xmrk kinase. AB - Extensive studies of primary tumors and tumor derived cell lines revealed that inappropriate activation of specific STATs (particularly of STAT3 and STAT5) occurs with high frequency in a wide variety of human cancers. We reported recently that the melanoma inducing EGFR-related receptor Xmrk specifically induces constitutive activation of STAT5 in fish melanoma cells. However, little is known about the role of STAT5 in solid tumours in general and its function in melanoma in particular. Recent examinations suggest that activated STAT signalling participates in oncogenesis by stimulating cell proliferation and preventing apoptosis. As an initial approach to understanding the consequences of Xmrk induced STAT5 signalling we used the well characterized pro B-cell line Ba/F3 as a sensitive system to analyse mitogenic as well as anti-apoptotic signalling. We identified STAT5 activation as being involved in both growth and survival signalling triggered by the Xmrk kinase possibly due to STAT5 induced expression of pim-1 and bcl-x. We also found a new mechanism of activation of STAT5 by receptor tyrosine kinases, whereby direct interaction of the receptor kinase domain with the STAT protein in a phosphotyrosine independent way led to activation of STAT5 in terms of DNA binding and target gene expression. PMID- 11896599 TI - Expression of the FUS domain restores liposarcoma development in CHOP transgenic mice. AB - Fusion proteins created by chromosomal abnormalities are key components of mesenchymal cancer development. The most common chromosomal translocation in liposarcomas, t(12;16)(q13;p11), creates the FUS-CHOP fusion gene. In the past, we generated FUS-CHOP and CHOP transgenic mice and have shown that while FUS-CHOP transgenic develop liposarcomas, mice expressing CHOP, which lacks the FUS domain, display essentially normal white adipose tissue (WAT) development, suggesting that the FUS domain of FUS-CHOP plays a specific and critical role in the pathogenesis of liposarcoma. To test the significance of FUS and CHOP domain interactions within a living mouse, we generated mice expressing the FUS domain and crossed them with CHOP-transgenic mice to generate double-transgenic FUSxCHOP animals. Here we report that expression of the FUS domain restores liposarcoma development in CHOP-transgenic mice. Our results provide genetic evidence that FUS and CHOP domains function in trans for the mutual restoration of liposarcoma. These results identify a new mechanism of tumor-associated fusion genes and might have impact beyond myxoid liposarcoma. PMID- 11896600 TI - p8-deficient fibroblasts grow more rapidly and are more resistant to adriamycin induced apoptosis. AB - p8 is a stress-induced DNA-binding protein, biochemically related to the architectural chromatin binding HMG protein family and whose function is presently unknown. We obtained fibroblast from mice lacking p8 and found that p8 is involved in cell growth regulation and in apoptosis. p8(-/-) mouse embryonic fibroblasts (MEFs) grow more rapidly than p8(+/+) MEFs. This might be explained by the higher intracellular level and activity of the Cdk2 and Cdk4 observed in p8(-/-) MEFs, which in turn may result, at least in part, from the concomitant decrease observed in the amount of cyclin-dependent kinase inhibitor p27. We also report that p8 mRNA expression is strongly activated in fibroblasts after cell growth arrest induced by serum deprivation or confluence. As expected, MEFs expressing p8 arrest their growth more rapidly after serum deprivation than MEFs lacking p8, which strongly suggests that p8 over-expression is implicated in cell growth arrest. On the other hand, p8(+/+) MEFs are more sensitive than p8(-/-) MEFs to the apoptosis induced by adriamycin treatment. p53 might be involved, as p8 expression increases its intracellular amount and trans-activation capacity. Finally, demonstration that p53 is a negative trans-activator of p8 suggests the presence of a complex autoregulatory loop. In conclusion, p8 is a cell growth inhibitor that facilitates apoptosis induced in fibroblasts by DNA damage. PMID- 11896601 TI - c-Src activation by the E5 oncoprotein enables transformation independently of PDGF receptor activation. AB - The E5 oncoprotein of bovine papillomavirus type 1 is a Golgi-resident, hydrophobic polypeptide that can transform immortalized fibroblasts by activating endogenous platelet-derived growth factor receptor beta (PDGF-R). However, the existence of E5 mutants that dissociate transformation from PDGF-R activation implies that there are additional mechanism(s) by which E5 can transform cells. We now show that both wt E5, and transforming E5 mutants that are defective for PDGF-R activation, constitutively activate endogenous c-Src in NIH3T3 cell lines to levels normally associated with acute growth factor stimulation. The ubiquitous Src family protein tyrosine kinase (PTK) Fyn is not activated by these E5 constructs, nor are focal adhesion kinase and endogenous receptor PTKs for insulin, epidermal growth factor, basic fibroblast growth factor and insulin-like growth factor. We further demonstrate that transforming activity of the L26A E5 mutant, which is highly defective for PDGF-R activation, depends on its ability to activate Src. L26A E5 does not transform SYF cells that are deficient for Src, Fyn and Yes, unless Src expression is reconstituted, and does not transform NIH3T3 cells in which Src PTK activity is maintained at a basal level by means of kinase-defective K295R Src overexpression. PMID- 11896602 TI - Membrane-anchored Cbl suppresses Hck protein-tyrosine kinase mediated cellular transformation. AB - The mammalian proto-oncogene Cbl and its cellular homologues in Caenorhabditis elegans (Sli-1) and Drosophila (D-Cbl) are negative regulators of some growth factor receptor signaling pathways. Herein we show that Cbl can negatively regulate another signaling molecule, namely theSrc-family kinase Hck by targeting it for degradation. Hck-mediated cellular transformation of murine fibroblasts is reverted by ectopic expression of a membrane-anchored allele of Cbl as assessed by the cellular morphology, suppression of anchorage independent growth, and an overall reduction in the total tyrosine phosphorylation levels within the cells. The expression of Cbl at the plasma membrane targets both Hck and itself for ubiquitination and degradation, requiring an intact RING finger. Pharmacological inhibition of the proteasome prevents the degradation of Hck correlating with an increase in the phosphotyrosine levels within the cells. Activated Hck and membrane-anchored Cbl are present in similar subcellular localizations and co immunoprecipitate, suggesting that their interaction is required for subsequent ubiquitination and degradation. Interestingly, both constitutively active and kinase-inactive Hck interact with and are targeted for degradation by Cbl. This work illustrates alternate means to regulate Src-family kinases, and suggests that Cbl may be able to suppress many signaling pathways that are activated in various proliferative syndromes including cancer. PMID- 11896603 TI - Cdc25C interacts with PCNA at G2/M transition. AB - Cdc25 activates maturation promoting factor (MPF) and promotes mitosis by removing the inhibitory phosphate from the Tyr-15 of Cdc2 in human cells. In this study, we searched the interacting protein(s) of human Cdc25C using the yeast two hybrid screen and identified proliferating cell nuclear antigen (PCNA) as an interacting partner of Cdc25C. The interaction between Cdc25C and PCNA was confirmed in vitro and in vivo. Co-immunoprecipitation analyses using human T cell line, Jurkat, further revealed that Cdc25C interacted with PCNA transiently when cells began to enter mitosis. Immunofluorescence analysis also showed that Cdc25C and PCNA were transiently co-localized in the nucleus at the beginning of M phase. Together with the previous observations of the interaction between various cdc/cyclin and PCNA, our findings strongly suggested a potential role of PCNA at the G2 to M phase transition of cell cycle. PMID- 11896604 TI - Interference with PDK1-Akt survival signaling pathway by UCN-01 (7 hydroxystaurosporine). AB - 3-Phosphoinositide-dependent protein kinase-1 (PDK1) plays a central role in activating the AGC subfamily of protein kinases. In particular, PDK1 plays an important role in the regulation of Akt/PKB survival pathway by phosphorylating Akt on Thr308. Here we show that UCN-01 (7-hydroxystaurosporine), a drug now in clinical trials and with a unique fingerprint pattern, induced dephosphorylation and inactivation of Akt, resulting in the turn-off of the survival signals and the induction of apoptosis. Further analysis revealed that UCN-01-mediated Akt inactivation was caused by inhibiting upstream Akt kinase PDK1 (IC50=33 nM) both in vitro and from cells, but not by suppressing Akt itself or phosphatidylinositide-3-OH kinase. UCN-01-induced PDK1 inhibition was also observed in in vivo murine and human tumor xenografts. Overexpression of active form of Akt diminished the cytotoxic effects of UCN-01, suggesting that UCN-01 may in part exert its cytotoxicity by inhibiting PDK1-Akt survival pathway. Because UCN-01 has already proved to have potent anti-tumor activity in vivo, PDK1-Akt survival pathway is a new, attractive target for cancer chemotherapy. PMID- 11896605 TI - Glial cell line-derived neurotrophic factor induces proliferative inhibition of NT2/D1 cells through RET-mediated up-regulation of the cyclin-dependent kinase inhibitor p27(kip1). AB - Growth factors of the glial cell line-derived neurotrophic factor (GDNF) family control the differentiation of neuronal cells of the central and peripheral nervous systems. Intracellular signalling of these growth factors is, at least in part, mediated by activation of the RET receptor tyrosine kinase. Here, we demonstrate that GDNF triggering inhibits the proliferation of the embryonal carcinoma cell line NT2/D1. This anti-proliferative effect is accompanied by down regulation of the SSEA-3 antigen, a marker typical of undifferentiated NT2/D1 cells. We show that these effects are mediated by activation of RET signalling. The block of RET by a kinase-deficient dominant negative mutant impairs GDNF dependent growth inhibition, whereas the adoptive expression of a constitutively active RET, the RET-MEN2A oncogene, promotes effects similar to those exerted by GDNF. We show that RET signalling increases the expression of the cyclin dependent kinase inhibitor p27(kip1) in NT2/D1 cells. Both DNA synthesis inhibition and SSEA-3 down-regulation are prevented if p27(kip1) expression is blocked by an antisense construct, which demonstrates that RET-triggered effects are mediated by p27(kip1). PMID- 11896606 TI - The subcellular localization of cyclin dependent kinase 2 determines the fate of mesangial cells: role in apoptosis and proliferation. AB - Apoptosis is closely linked to proliferation. In this study we showed that inducing apoptosis in mouse mesangial cells with ultraviolet (UV) irradiation was associated with increased cyclin A-cyclin dependent kinase (CDK) 2 activity. Inhibiting CDK2 activity with Roscovitine or dominant negative mutant reduced apoptosis. Because apoptosis typically begins in the cytoplasm, we tested the hypothesis that the subcellular localization of CDK2 determines the proliferative or apoptotic fate of the cell. Our results showed that cyclin A-CDK2 was nuclear in proliferating cells. However, inducing apoptosis in proliferating cells with UV irradiation was associated with a decrease in nuclear cyclin A and CDK2 protein levels. This coincided with an increase in protein and kinase activity for cyclin A-CDK2 in the cytoplasm. Translocation of cyclin A-CDK2 also occurred in p53-/- mesangial cells. Finally, we showed that caspase-3 activity was significantly reduced by inhibiting CDK2 activity with Roscovitine. In summary, our results show that apoptosis is associated with an increase in cytoplasmic cyclin A-CDK2 activity, which is p53 independent and upstream of caspase-3. We propose that the subcellular localization of CDK2 determines the proliferative or apoptotic fate of the cell. PMID- 11896607 TI - Silibinin inhibits constitutive and TNFalpha-induced activation of NF-kappaB and sensitizes human prostate carcinoma DU145 cells to TNFalpha-induced apoptosis. AB - Prostate cancer (PCA) is one of the most common invasive malignancies of men in the US, however, there have been limited successes so far in its therapy. Even most potent agents (e.g. TNFalpha) are ineffective in killing human PCA cells possibly due to constitutive activation of NF-kappaB that subsequently activates a large number of anti-apoptotic genes. In such a scenario, strong apoptotic agent TNFalpha, further induces NF-kappaB activation rather than inducing apoptosis. In several recent studies, we have demonstrated both cancer preventive and anti-cancer efficacy of silymarin and its constituent silibinin in a variety of experimental tumor models and cell culture systems. Here we examined whether silibinin is effective in inhibiting constitutive NF-kappaB activation in human PCA cells, which would help in overcoming TNFalpha-insensitivity. Our studies reveal that silibinin effectively inhibits constitutive activation of NF-kappaB in advanced human prostate carcinoma DU145 cells. Consistent with this, nuclear levels of p65 and p50 sub-units of NF-kappaB were also reduced. In the studies assessing molecular mechanism of this effect, silibinin treatment resulted in a significant increase in the level of IkappaBalpha with a concomitant decrease in phospho-IkappaBalpha. Kinase assays revealed that silibinin dose-dependently decreases IKKalpha kinase activity. The effect of silibinin on IKKalpha seemed to be direct as evidenced by the in vitro kinase assay, where immunoprecipitated IKKalpha was incubated with silibinin. This shows that silibinin does not necessarily need an upstream event to bring about its inhibitory effect on IKKalpha and downstream effectors. Additional studies showed that silibinin also inhibits TNFalpha-induced activation of NF-kappaB via IkappaBalpha pathway and subsequently sensitizes DU145 cells to TNFalpha-induced apoptosis. These results indicate that silibinin could be used to enhance the effectiveness of TNFalpha based chemotherapy in advanced PCA. PMID- 11896608 TI - Multiple mutations are common at mouse Aprt in genotoxin-exposed mismatch repair deficient cells. AB - Mismatch repair deficiency is known to contribute to elevated rates of mutations, particularly at mono- and dinucleotide repeat sequences. However, such repeats are often missing from the coding regions of endogenous genes. To determine the types of mutations that can occur within an endogenous gene lacking highly susceptible repeat sequences, we examined mutagenic events at the 2.3 kb mouse Aprt gene in kidney cell lines derived from mice deficient for the PMS2 and MLH1 mismatch repair proteins. The Aprt mutation rate was increased 33-fold and 3.6-20 fold for Mlh1 and Pms2 null cell lines, respectively, when compared with a wild type kidney cell line. For the Pms2 null cells this increase resulted from both intragenic events, which were predominantly base-pairs substitutions, and loss of heterozygosity events. Almost all mutations in the Mlh1 null cells were due to base-pair substitutions. A:T-->G:C transitions (54% of small events) were predominant in the Pms2 null cells whereas G:C-->A:T transitions (36%) were the most common base-pair change in the Mlh1 null cells. Interestingly, 4-9% of the spontaneous mutant alleles in the mismatch repair deficient cells exhibited two well-separated base-pair substitution events. The percentage of mutant alleles with two and occasionally three base-pair substitutions increased when the Pms2 and Mlh1 null cells were treated with ultraviolet radiation (15-21%) and when the Mlh1 null cells were treated with hydrogen peroxide (35%). In most cases the distance separating the multiple base-pair substitutions on a given allele was in excess of 100 base-pairs, suggesting that the two mutational events were not linked directly to a single DNA lesion. The significance of these results is discussed with regards to the roles for the PMS2 and MLH1 proteins in preventing spontaneous and genotoxin-related mutations. PMID- 11896609 TI - Cdc6 requires anchorage for its expression. AB - Fibroblasts need anchorage to extracellular matrix to transit from G1 to S phase, but no longer after oncogenic transformation. Here we report that Cdc6 protein essential for the activation of replication origins requires anchorage or oncogenic stimulation for its execution. Upon anchorage loss, Cdc6 expression is shut off both transcriptionally and post-transcriptionally in a rat fibroblast despite enforced activation of E2F-dependent promoters. However, stimulation of this cell with oncogenic growth factors suppresses this shutoff and concurrently activates Cdk2 and Cdk6/4, thereby overriding the anchorage requirement for the G1-S transition and consequently enabling cells to perform anchorage-independent S phase entry. Analysis with enforced expression of Cdc6 indicates that the G1 cyclin-dependent kinases and Cdc6 constitute major cell cycle targets for the restriction of the G1-S transition by anchorage loss. PMID- 11896610 TI - Feline STK gene expression in mammary carcinomas. AB - The human RON and its mouse homologue stk are members of the MET family of tyrosine kinase receptors. We have previously shown that the RON gene is over expressed in human breast carcinomas. As cat mammary tumours have been proposed as a suitable model for aggressive human breast cancer, we identified the feline stk gene and studied its expression in cat mammary cancer. Feline stk sequences were found highly homologous to the stk and RON gene exons that encode the juxtamembrane and transmembrane domains of the stk and RON receptors. Feline stk specific transcript was detected by RT-PCR in cat lung and in 7/8 feline mammary carcinomas and a synchronous skin metastasis examined. Western blot and immunohistochemical analyses were carried out with an antibody that recognized both the human RON and mouse stk receptors. This antibody specifically detected a 135 Kd feline protein and stained 10/34 mammary carcinoma archival samples. These data show that the pattern of expression and distribution of the stk protein in feline mammary cancer could be superimposed on that of the RON receptor in human breast cancer and suggest that these feline tumours are a suitable model to test innovative approaches to therapy of aggressive human breast carcinomas. PMID- 11896611 TI - Hepatocyte growth factor promotes hepatocarcinogenesis through c-Met autocrine activation and enhanced angiogenesis in transgenic mice treated with diethylnitrosamine. AB - Hepatocyte growth factor (HGF) is a mitogen for hepatocytes, but it is not clear whether HGF stimulates or inhibits hepatocarcinogenesis. We previously reported that HGF transgenic mice under the metallothionein gene promoter developed benign and malignant liver tumors spontaneously after 17 months of age. To elucidate the role of HGF in hepatocarcinogenesis, diethylnitrosamine (DEN) was administered to HGF transgenic mice. HGF overexpression accelerated DEN-induced hepatocarcinogenesis, often accompanied by abnormal blood vessel formation. In this study, 59% of transgenic males (versus 20% of wild-type males) and 39% of transgenic females (versus 2% of wild-type females) developed either benign or malignant liver tumors by 48 weeks (P<0.005, P<0.001, respectively). Moreover, 33% of males and 23% of female transgenic mice developed hepatocellular carcinoma (HCC), while none of the wild-type mice developed HCC (P<0.001, P<0.005, respectively). Enhanced kinase activity of the HGF receptor, Met, was detected in most of these tumors. Expression of vascular endothelial growth factor (VEGF) was up-regulated in parallel with HGF transgene expression. Taken together, our results suggest that HGF promotes hepatocarcinogenesis through the autocrine activation of the HGF-Met signaling pathway in association with stimulation of angiogenesis by HGF itself and/or indirectly through VEGF. PMID- 11896612 TI - Use of signal specific receptor tyrosine kinase oncoproteins reveals that pathways downstream from Grb2 or Shc are sufficient for cell transformation and metastasis. AB - Many human cancers have been associated with the deregulation of receptor tyrosine kinases (RTK). However, the individual contribution of receptor associated signaling proteins in cellular transformation and metastasis is poorly understood. To examine the role of RTK activated signal transduction pathways to processes involved in cell transformation, we have exploited the oncogenic derivative of the Met RTK (Tpr-Met). Unlike other RTKs, twin tyrosine residues in the carboxy-terminal tail of the Met oncoprotein and receptor are required for all biological and transforming activities, and a mutant lacking these tyrosines is catalytically active but non transforming. Using this mutant we have inserted oligonucleotide cassettes, each encoding a binding site for a specific signaling protein derived from other RTKs. We have generated variant forms of the Tpr-Met oncoprotein with the ability to bind individually to the p85 subunit of PI3'K, PLCgamma, or to the Grb2 or Shc adaptor proteins. Variants that recruit the Shc or Grb2 adaptor proteins generated foci of morphologically transformed fibroblast cells and induced anchorage-independent growth, scattering of epithelial cells and experimental metastasis. In contrast, variants that bind and activate PI3'K or PLCgamma failed to generate readily detectable foci. Although cell lines expressing the PI3'K variant grew in soft-agar, these cells were non metastatic. Using this unique RTK oncoprotein model, we have established that Grb2 or Shc dependent signaling pathways are sufficient for cell transformation and metastatic spread. PMID- 11896613 TI - Constitutive expression of the Id-1 promoter in human metastatic breast cancer cells is linked with the loss of NF-1/Rb/HDAC-1 transcription repressor complex. AB - The helix-loop-helix protein Id-1 is a dominant negative regulator of basic helix loop-helix transcription factors, and plays a key role in the control of breast epithelial cell growth, invasion and differentiation. Previous investigations in our laboratory have shown that Id-1 mRNA was constitutively expressed in highly aggressive and invasive human breast cancer cells in comparison to non transformed or non-aggressive cancerous cells, and that this loss of regulation is mediated by a 2.2-kb region of the human Id-1 promoter. Here we show that a 31 bp sequence within this 2.2-kb promoter, located 200 bp upstream of the initiation of transcription, is responsible for the constitutive expression of Id 1 in metastatic human breast cancer cells. Using gel shift experiments, we identified a high molecular weight complex present only in non-aggressive breast cancer cells cultured in serum-free medium and which appear to be necessary for proper Id-1 repression. In contrast, nuclear extracts from highly aggressive and metastatic cell lines do not contain this large molecular weight complex. Using DNA affinity precipitation assays (DAPA), we show that this complex contains SP 1, NF-1, Rb and HDAC-1 proteins. On the basis of these findings, we propose a mechanism for the loss of regulation of Id-1 promoter in invasive and metastatic human breast cancer cells. PMID- 11896614 TI - p16INK4a loss and sensitivity in KSHV associated primary effusion lymphoma. AB - The Kaposi's Sarcoma associated Herpes virus (KSHV) encodes two genes with the potential to affect the activity of the retinoblastoma protein (Rb). Open reading frame (orf) 72 encodes a D type cyclin (kcyc) that can elicit p16INK4a resistant cdk activity and orf73 encodes the latency associated nuclear antigen (LNA) that can bind Rb and neutralize E2F regulation. This indicates that, like papilloma and adenovirus associated malignancies, those associated with KSHV are defective with respect to their Rb pathway. To address this we investigated whether KSHV associated primary effusion lymphoma (PEL) derived cell lines are resistant to growth inhibition by p16INK4a. We provide evidence that ectopic expression of p16INK4a in these cells causes an Rb dependent G1 cell cycle block. Importantly, endogenous p16INK4a expression is not detected in six PEL derived cell lines and four primary PEL samples and examination of the p16INK4a locus shows deletion in two out of six and hypermethylation in four out of six PEL lines. Treatment of the latter with the demethylating agent 5'-aza-2' deoxycytidine leads to re expression of p16INK4a protein. Taken together these results suggest that p16INK4a loss may be a cellular change frequently associated with PEL. They furthermore argue that despite the presence of KSHV DNA and expression of a latent gene program Rb function is intact in PEL. PMID- 11896615 TI - Alternative transcripts of the candidate tumor suppressor gene, WWOX, are expressed at high levels in human breast tumors. AB - The presence of putative tumor-suppressor genes on chromosome 16q23.2-24.1 has been suggested by LOH analysis in several cancer types. This region overlaps with the fragile site FRA16D and the region of homozygous deletions found in several cancer types. The candidate gene WWOX/FOR has been mapped within this region. The mouse homologue of the WWOX protein has been defined as an apoptogenic protein and an essential partner of p53 in cell death, supporting WWOX as a tumor suppressor gene candidate. We performed an expression study of the WWOX/FOR gene in a series of human breast tumors and breast cancer cell lines, and detected reduced expression of the WWOX/FOR transcript in a series of breast cancer cells. Furthermore, identification of two distinct alternative WWOX transcripts expressed at high levels in human tumors suggests an involvement of the WWOX gene in breast cancer progression. PMID- 11896616 TI - Absence of fibroblast growth factor 2 does not prevent tumor formation originating from the RPE. AB - We have analysed the importance of fibroblast growth factor 2 (FGF2) in tumor development. In a transgenic mouse model (Tyrp1-Tag) tumors form in the retinal pigment epithelium (RPE), invade surrounding tissues, and metastasize to lymph node and spleen. To address whether RPE tumor formation is dependent on FGF2, we generated FGF2-deficient mice. Such mice appeared healthy and exhibited no impairment of growth or development. Tyrp1-Tag transgenic mice, which are lacking FGF2 (FGF2-/-) developed RPE tumors that metastasize to spleen and lymph nodes. Tumor growth and survival rate are identical to Tyrp1-Tag transgenic littermates expressing FGF2. Cell lines were isolated from RPE tumors of wild-type and FGF2 deficient mice. They grow in culture, are pigmented and form vascularized tumors, when injected subcutaneously into nude mice of either FGF2-/- or FGF2+/+ genetic background. Kinetics of tumor growth was identical and independent of presence of FGF2. Together, these results demonstrate that FGF2 is not essential for tumor formation of the RPE thus suggesting that tumor growth in general may not be dependent on FGF2. PMID- 11896617 TI - Caspase-9 and Apaf-1 are expressed and functionally active in human neuroblastoma tumor cell lines with 1p36 LOH and amplified MYCN. AB - Important roles have been suggested for caspase-8, caspase-9 and Apaf-1 in controlling tumor development and their sensitivity to chemotherapeutic agents. Methylation and deletion of Apaf-1 and CASP8 results in the loss of their expression in melanoma and neuroblastoma, respectively, while CASP9 localization to 1p36.1 suggests it is a good candidate tumor suppressor. The status of CASP9 and Apaf-1 expression in numerous neuroblastoma cell lines with/without amplified MYCN and chromosome 1p36 loss-of-heterozygosity (LOH) was therefore examined to test the hypothesis that one or both of these genes are tumor suppressors in neuroblastoma. Although CASP9 is included in the region encompassing 1p36 LOH in all neuroblastoma cell lines examined, the remaining CASP9 allele(s) express a functional caspase-9 enzyme. Apaf-1 is also expressed in all neuroblastoma tumor cell lines examined. Thus, the CASP9 or Apaf-1 genes do not appear to function as tumor suppressors in MYCN amplified neuroblastomas. However, approximately 20% of the neuroblastoma cell lines with methylated CASP8 alleles are also highly resistant to staurosporine (STS)- and radiation-induced cell death, presumably because cytochrome c is not released from mitochondria. This suggests that a second, smaller sub-group of MYCN amplified neuroblastoma tumors exists with defect(s) in apoptotic signaling components upstream of caspase-9 and Apaf-1. Since no consistent differences in Bcl-2, Bcl-x(L) or Bax expression were seen in the STS- and radiation-resistant neuroblastomas, it suggests that a unique mitochondrial signaling factor(s) is responsible for the defect in cytochrome c release in this sub-group of tumors. PMID- 11896619 TI - SHP-2 is involved in heterodimer specific loss of phosphorylation of Tyr771 in the PDGF beta-receptor. AB - We have previously shown that the binding site for GTPase activating protein of Ras (RasGAP) in the PDGF beta-receptor, Tyr771, is phosphorylated to a much lower extent in the heterodimeric configuration of PDGF alpha- and beta-receptors, compared to the PDGF beta-receptor homodimer. The decreased recruitment of the RasGAP to the receptor leads to prolonged activation of the Ras/MAP kinase pathway, which could explain the increase in mitogenicity seen upon induction of heterodimers. The molecular mechanism underlying these differences was investigated. We could show that the loss of phosphorylation of Tyr771 was dependent on presence of intact binding sites for the protein tyrosine phosphatase SHP-2 on the PDGF beta-receptor. Thus, in PDGF receptor mutants in which binding of SHP-2 was lost, a higher degree of phosphorylation of Tyr771 was seen, while other phosphorylation sites in the receptor remained virtually unaffected. Thus, SHP-2 appears to play an important role in modulating phosphorylation of Y771, thereby controlling RasGAP recruitment and Ras/MAP kinase signaling in the heterodimeric configuration of the PDGF receptors. PMID- 11896618 TI - A Myb dependent pathway maintains Friend murine erythroleukemia cells in an immature and proliferating state. AB - Friend murine erythroleukemia (MEL) cells are transformed erythroid precursors that are held in an immature and proliferating state but can be induced to differentiate in vivo by treatment with a variety of chemical agents such as N, N hexamethylene bisacetamide (HMBA). To investigate the role of Myb proteins in maintaining MEL cells in an immature and proliferating state we have produced stable transfectants in the C19 MEL cell line that contain a dominant interfering Myb allele (MEnT) under the control of an inducible mouse metallothionein I promoter. When expression of MEnT protein was induced with ZnCl2, the stable transfectants differentiated with kinetics that were similar to wild type C19 MEL cells treated with HMBA, including induction of alpha-globin mRNA expression, assembly of hemoglobin and growth arrest. Expression of endogenous c-myb and c myc was also decreased in response to MEnT. Expression of mad-1 mRNA was rapidly increased in response to expression of MEnT resulting in a shift from predominantly c-Myc/Max complexes to predominantly Mad/Max containing complexes. These results strongly suggest that C19 MEL cells are held in an immature and proliferating state by a pathway that is dependent on Myb activity. PMID- 11896620 TI - Coincident inactivation of 14-3-3sigma and p16INK4a is an early event in vulval squamous neoplasia. AB - The structure and expression of 14-3-3 sigma(sigma) was analysed in squamous carcinomas (SCC) of the vulva and in the vulval pre-malignant lesion vulval intraepithelial neoplasia (VIN). Sequence analysis of the sigma coding region did not detect mutations in any case of SCC or VIN III and loss of heterozygosity (LOH) occurred in only 2 out of 27 informative cases. In contrast to the absence of genetic change, methylation-specific PCR (MSP) analysis revealed dense CpG methylation within the sigma gene in approximately 60% of cases of vulval SCC, but methylation was not detected in matched, normal epithelial tissue. Methylation was associated in all cases with reduced or absent expression of sigma mRNA. There was no correlation between sigma methylation and HPV or p53 status. Analysis of pre-malignant vulval intraepithelial neoplasia (VIN) revealed that sigma methylation was detectable early in neoplastic development. Co incident methylation, accompanied by loss of expression, of sigma and p16INK4a was commonly detected in both SCC and VIN III, suggesting that epigenetic silencing of these two genes is an early and important event in vulval neoplasia. PMID- 11896622 TI - Transcriptional regulation of IGF-I receptor gene expression by novel isoforms of the EWS-WT1 fusion protein. AB - The EWS family of genes is involved in numerous chromosomal translocations that are characteristic of a variety of sarcomas. A recently described member of this group is desmoplastic small round cell tumor (DSRCT), which is characterized by a recurrent t(11;22)(p13;q12) translocation that fuses the 5' exons of the EWS gene to the 3' exons of the WT1 gene. The originally described chimera comprises exons 1-7 of EWS and exons 8-10 of WT1. We have previously reported that the WT1 protein represses the expression of the IGF-I receptor gene, whereas the EWS(1-7) WT1(8-10) fusion protein activates IGF-I receptor gene expression. It has recently become apparent that EWS-WT1 chimeras produced in DSCRT are heterogeneous as a result of fusions of different regions of the EWS gene to the WT1 gene. We have recently characterized additional EWS-WT1 translocations that involve the juxtaposition of EWS exons 7 or 8 to WT1 exon 8, and an EWS-WT1 chimera that lacks EWS exon 6. The chimeric transcription factors encoded by these various translocations differ in their DNA-binding characteristics and their ability to transactivate the IGF-I receptor promoter. These data suggest that the molecular pathology of DSRCT is more complex than previously appreciated, and that this diversity may provide the foundation for predictive genotype-phenotype correlations in the future. PMID- 11896623 TI - Ectopic expression of FGF-3 results in abnormal prostate and Wolffian duct development. AB - To evaluate the effects of FGF-3 expression in the prostate and male reproductive tract, we employed a bitransgenic system to target FGF-3 to these organs. We present a first study that ectopic FGF-3 expression resulted in exuberant hyperplasia of all bigenic prostatic lobes typified by epithelial stratification, cribiform structures and papillary tufts. These cells displayed increased nuclear to-cytoplasmic ratios and bromodeoxyuridine (BrdU) proliferative index but retained relatively uniform nuclear androgen receptor (AR) and the tumor suppressor C-CAM1 staining. Furthermore, the dysmorphogenic prostatic cells also resembled PIN (prostatic intraepithelial neoplasia)-like lesions but did not appear to have invaded the basal lamina. In addition to these phenotypes, profound disorders of the bigenic Wolffian duct derivatives were observed. The bigenic ampullary glands and vas deferens were extremely cystic, hypertrophic and hyperplastic; the enlarged epididymi showed a reduction of spermatozoa and the seminal vesicles exhibited a dramatic reduction of seminal secretions. Because of these severe abnormalities, these infertile males presented with diaphragmatic hernias, hemoperitoneum and many secondary abnormalities at sacrifice. Taken together, we show that ectopic FGF-3 expression severely perturbs normal prostate development and our system should be useful for the analyses of early changes in prostatic hyperplasia. PMID- 11896621 TI - Inositol hexakisphosphate kinase 2 sensitizes ovarian carcinoma cells to multiple cancer therapeutics. AB - We recently identified inositol hexakisphosphate kinase 2 (IP6K2) as a positive regulator of apoptosis. Overexpression of IP6K2 enhances apoptosis induced by interferon-beta (IFN-beta) and cytotoxic agents in NIH-OVCAR-3 ovarian carcinoma cells. In this study, we contrast and compare IFN-beta and radiation-induced death, and show that IP6K2 expression sensitizes tumor cells. Unirradiated NIH OVCAR-3 cells transfected with IP6K2 formed fewer colonies compared to unirradiated vector-expressing cells. IP6K2 overexpression caused increased radiosensitivity, evidenced by decreased colony forming units (CFU). Both IFN beta and radiation induced caspase 8. IFN-beta, but not gamma-irradiation, induced TRAIL in NIH-OVCAR-3 cells. Gamma irradiation, but not IFN-beta, induced DR4 mRNA. Apoptotic effects of IFN-beta or gamma-irradiation were blocked by expression of a dominant negative mutant death receptor 5 (DR5Delta) or by Bcl-2. Caspase-8 mRNA induction was more pronounced in IP6K2-expressing cells compared to vector-expressing cells. These data suggest that overexpression of IP6K2 enhances sensitivity of some ovarian carcinomas to radiation and IFN-beta. IP6K2 may function to enhance the expression and/or function of caspase 8 and DR4 following cell injury. Both IFN-beta and gamma-irradiation induce apoptosis through the extrinsic, receptor-mediated pathway, IFN-beta through TRAIL, radiation through DR4, and both through caspase 8. The function of both death inducers is positively regulated by IP6K2. PMID- 11896624 TI - An endonuclease/ligase based mutation scanning method especially suited for analysis of neoplastic tissue. AB - Knowledge of inherited and sporadic mutations in known and candidate cancer genes may influence clinical decisions. We have developed a mutation scanning method that combines thermostable EndonucleaseV (Endo V) and DNA ligase. Variant and wild-type PCR amplicons are generated using fluorescently labeled primers, and heteroduplexed. Thermotoga maritima (Tma) EndoV recognizes and primarily cleaves heteroduplex DNA one base 3' to the mismatch, as well as nicking matched DNA at low levels. Thermus species (Tsp.) AK16D DNA ligase reseals the background nicks to create a highly sensitive and specific assay. The fragment mobility on a DNA sequencing gel reveals the approximate position of the mutation. This method identified 31/35 and 8/8 unique point mutations and insertions/deletions, respectively, in the p53, VHL, K-ras, APC, BRCA1, and BRCA2 genes. The method has the sensitivity to detect K-ras mutations diluted 1 : 20 with wild-type DNA, a p53 mutation in a 1.7 kb amplicon, and unknown p53 mutations in pooled DNA samples. EndoV/Ligase mutation scanning combined with PCR/LDR/Universal array proved superior to automated DNA sequencing for detecting p53 mutations in colon tumors. This technique is well suited for scanning low-frequency mutations in pooled samples and for analysing tumor DNA containing a minority of the unknown mutation. PMID- 11896625 TI - A non-transgenic mouse model for B-cell lymphoma: in vivo infection of p53-null bone marrow progenitors by a Myc retrovirus is sufficient for tumorigenesis. AB - The c-Myc oncoprotein is strongly implicated in B-cell neoplasms such as human Burkitt lymphomas and mouse plasmocytomas. Transgenic mice in which the myc gene is juxtaposed to an immunoglobulin enhancer (E(mu)-myc) also develop B-cell lymphomas, but relatively late in life. In addition, these neoplasms are invariably clonal, suggesting the involvement of additional mutations. Such mutations frequently affect the p53 tumour suppressor gene or its positive regulator Arf, hinting that inactivation of the p53 pathway might be the second hit required for the progression towards malignancy. However, even tumours arising in E(mu)-myc/Arf-null animals are thought to be clonal. This observation raised doubts whether overexpression of Myc in p53-null B-cell precursors is sufficient for tumorigenesis. To address this question, we have established a new, non-transgenic mouse model of B-lymphoma. This model is based on isolation of primary bone marrow (BM) cells, admixing them with packaging cells producing a Myc-encoding retrovirus (LMycSN), and subcutaneous injection into a host with which BM cells are syngeneic. Predictably, wild type BM cells infected in vivo by LMycSN were not tumorigenic. However, LMycSN-infected p53-null BM cells readily gave rise to B-cell lymphomas composed predominantly of late pro-B/small pre-B cells. In these tumours, heavy chain gene rearrangements were analysed using two independent PCR-based assays. All neoplasms with DJ-rearrangements were found to be polyclonal. This result suggests that inactivation of p53 and overexpression of Myc is all that is necessary for the development of full-fledged B-lymphomas. Our model would also be instrumental in assessing the transforming potential of Myc mutants and in studying cooperation between Myc and other oncogenes. PMID- 11896626 TI - Contribution of cyclin d1 (CCND1) and E-cadherin (CDH1) polymorphisms to familial and sporadic colorectal cancer. AB - The molecular basis for most non-HNPCC familial colorectal cancer cases is unknown, but there is increasing evidence that common genetic variants may play a role. We investigated the contribution of polymorphisms in two genes implicated in the pathogenesis of colorectal cancer, cyclin D1 (CCND1) and E-cadherin (CDH1), to familial and sporadic forms of the disease. The CCND1 870A/G polymorphism is thought to affect the expression of CCND1 through mRNA splicing and has been reported to modify the penetrance of HNPCC. Inactivation of E cadherin is common in colorectal cancer, and truncating germline mutations have been reported to confer susceptibility to colorectal as well as diffuse gastric cancer. The -160A/C CDH1 polymorphism appears to affect expression of CDH1 and may therefore also confer an increased risk. We found a significantly higher frequency of CCND1 870A allele in 206 familial cases compared to 171 controls (P=0.03). Odds ratios in heterozygotes and homozygotes were 1.7 (95% CI: 1.0 2.66) and 1.8 (95% CI: 1.0-3.3) respectively. The difference was accounted for by an over-representation of A allele in non-HNPCC familial cases (P=0.007). Over representation of the CCND1 A allele was also seen in sporadic colorectal cancer cases compared to controls but this did not attain statistical significance (P=0.08). No significant differences between the frequency of CDH1 -160A/C genotypes in familial, sporadic colorectal cancer cases and controls were seen, although a possible association between the low expressing A allele and right sided tumours was detected in familial cases. PMID- 11896627 TI - Gene expression profiling of HeLa cells in G1 or G2 phases. AB - The cell division cycle is regulated through both transcriptional and post transcriptional mechanisms. The altered expression of a number of genes at the mRNA level is known to be essential for progression through the cell cycle, however, a comprehensive gene expression profile of human cells remains to be completed. Here we sought to monitor the differential gene expression of genes after the transition of G2 cells into G1 prior to the restriction point. GeneChip containing microarrays of oligonucleotides corresponding to over 12 000 human genes were employed to profile differential gene expression in G1 and G2. After three independent experiments the resultant data was filtered and a set of genes was compiled based on at least threefold-altered expression, no background noise in determining expression and observation in all experiments. Our analysis identified 154 genes that were elevated in G2 phase of cells as compared to early G1 phase including 15 novel genes. This number included mRNAs whose upregulation is known to occur in G2 phase. Surprisingly only 19 genes were upregulated in G1 phase, among these six genes were novel. Some of these genes are candidates for transition through early G1. This gene inventory for G1 and G2 phases of cell cycle will provide the basis for understanding of cell cycle delay as a result of DNA damage. PMID- 11896629 TI - Planning for the unthinkable. PMID- 11896630 TI - Mass casualty chemical incidents--towards guidance for public health management. AB - The aim of this paper is two-fold. A review of mass casualty chemical incidents occurring naturally or as a result of industrial activities or deliberate release provided an opportunity to consider the problems experienced in medical and public health response. In addition, a literature review of procedures to assist in the management of chemical incidents by medical and public health professionals was conducted, targeted at summarising what tools exist to minimise the impact on public health from such events. We found that most of the large chemical incidents worldwide were unprecedented and unexpected, and as a consequence emergency services, hospitals and public health had access to very little relevant information in the first few hours. Several lessons were highlighted that would aid future response. The review of procedures showed that there are currently no written procedures that are both sufficiently generic and sufficiently detailed to effectively support the public health management of the emergency response to chemical incidents. There is therefore a need to fill this obvious gap and to develop a procedural guide for the emergency management of chemical incidents by public health professionals. This work is now under way. PMID- 11896631 TI - Violence in the community: a health service view from a UK Accident and Emergency Department. AB - A retrospective analysis of information recorded on victims of assault, who attended the Accident and Emergency department of Chorley and South Ribble Hospital over a 1 y period, was performed in order to describe the epidemiology of violent assault. During the year 735 (1.7%) of the patients attending A&E were identified as being victims of assault (71% were male). Victims were predominantly in their late teens and early 20s (median age 23 y, inter-quartile range 17 to 32 y). They attended at weekends (44% on Saturday or Sunday) and predominantly between the hours of 8 pm and 4 am (54%). Minor injuries were the most frequent (43%) while 21% of victims sustained lacerations and 11% had a fracture. The commonest site of injury was to the neck, face and throat (55%). The crude rate of attendance following violent assault for Chorley District was 4.67 per 1000 population per year. Information routinely collected by A&E departments can be used to describe the epidemiology of violence in the community. Further work is required to ascertain the optimal method of collection and dissemination of this information. PMID- 11896632 TI - Can school-related factors predict future health behaviour among young adolescents? AB - Although the prevention of unfavourable health behaviour among young people has high priority in public health, the possibility of finding risk factors at school has not been sufficiently studied. The objective of this study was to find predictors among young pupils for later unfavourable health behaviour, with special focus on school-related factors.A three-year prospective study was started in 1994 including 279 pupils (141 girls and 138 boys) from different socio-economic areas. The pupils answered a comprehensive questionnaire in grade six and grade nine. The non-response rate was negligible. The best predictors for health behaviour among boys and girls in grade nine were factors related to earlier health/health behaviour. The results also indicated that school-related factors could predict future health behaviour, especially in relation to low physical activity among girls. The school has an important role to play in identification of future unfavourable health behaviour among pupils at the school, both directly through recognising school-related risk factors and also indirectly through paying special attention to pupils with unfavourable health/health behaviour. Our findings indicate the need for more research in younger ages, as negative health behaviour already seems to be established at 12 y of age. PMID- 11896633 TI - Adolescent screening for orthopedic problems in high school. AB - BACKGROUND: The effectiveness of orthopedic screening programs for school-age children are still controversial. We conducted a prospective study in order to determine the frequency of undiagnosed orthopedic problems in an adolescent population. OBJECTIVE: To determine the frequency of undiagnosed orthopedic problems in an adolescent population discovered through routine physical examinations carried out by a general pediatrician in a school clinic. METHODS: We examined 2380 adolescents attending a public high school over a 5 y period in order to determine the frequency of undiagnosed orthopedic abnormalities in this age group. RESULTS: Previously undiagnosed orthopedic findings, especially spinal deformities were found in 14.8%. Scoliosis was detected in 1.6% of the entire group with a threefold predominance of girls over boys. Few cases were progressive and needed surgery. Extra spinal orthopedic findings were found in 2.9% of the patients. CONCLUSIONS: Screening programs can identify previously undetected orthopedic abnormalities in the school-age population. We conclude that screening programs for school age children coupled with subsequent follow-up procedures are worthwhile. PMID- 11896634 TI - Low birth weight is associated with elevated serum lipoprotein(a) in white and black American children ages 5-11 y. AB - Nutritional insults experienced by the mother have a life-long imprint on organ size and function of the fetus. Infant low birth weight (LBW) is one of the consequences of such maternal undernutrition. The physiological consequences of nutritional insults can lead to adverse metabolic consequences after birth, including hypertension, diabetes and dyslipidemia. The objective of this study was to determine the association and contribution of LBW to serum concentration of lipoprotein(a) [Lp(a)] in a representative sample of white and black American children aged 5-11 y. Data (n=666) from the Third US National Health and Nutrition Examination Survey were used in this investigation. Racial/ethnic specific trends in mean values of Lp(a) were compared across tertile distribution of birth weight. Multiple linear regression analysis was employed to determine the association of birth weight with Lp(a), controlling for age, sex and sum of four skinfold thicknesses (SUM). A consistent trend of increasing values of Lp(a) with decreasing birth weight emerged for both white and black children (P<0.001). Black children presented with higher values of serum concentrations of Lp(a) at each level of birth weight distributions than white children (P<0.01). Black race/ethnicity was associated with approximately 0.4 mg/dl greater serum concentration of Lp(a) than white, adjusting for birth weight, age, sex and SUM (P<0.001). LBW sub-population in black children appeared to be relevant to elevated Lp(a) concentration, while a similar scenario did not appear in white children. Since interactions between in utero factors and risk exposures after birth are likely, definitive studies evaluating these interactions are warranted. PMID- 11896635 TI - Stomach cancer-related mortality rate is higher in young Japanese women than in men. AB - This study compares stomach cancer-related mortality rates in Japan with those in European and Asian countries and analyzes trends in stomach cancer-related mortality rates according to gender in young Japanese over the period of 1957 1997. From official death certification numbers and population estimates, we obtained stomach cancer-related mortality rate for all ages and various ages according to gender. Japan's ranking compared to other countries in death percentage of all cancers which are attributable to stomach cancer was fourth for both men and women. In Japan and Ireland, total elimination of deaths from stomach cancer in men resulted in increased life expectancy of 0.68 and 0.22 y respectively, whereas the corresponding figures for women were 0.42 and 0.14 y respectively. The sex ratios of stomach cancer-related mortality rates were 0.75, 0.63, 0.80 and 0.94 for 25-29, 30-34, 35-39 and 40-44 y age groups, respectively, in 1997. The sex ratio of relative risk ranged from 0.62 to 0.92 in 25-40 y age groups during the observation period. The life expectancy in 30-34 y age group increased by 0.66 y for men and 0.41 y for women in 1995 after elimination of stomach cancer-related deaths. Our results suggest that stomach cancer-related mortality rates are still high in Japan and young women are at higher risk of stomach cancer-related death relative to young men and that sex ratio is stable or slightly decreased over the 40-y period. It is important to monitor this trend continuously in the next few years. PMID- 11896636 TI - Population based study of hypertension among the elderly in northern India. AB - The study was conducted in the urban and rural areas of Chandigarh during 1998 1999 among 362 elderly subjects above 65 y of age. The sample, selected by stratified random technique, covered a population of 7937 family members in 1882 houses. Methodology comprised of interviews, clinical examination and laboratory investigations such as ECG. The study revealed that 210 (58%) of the elderly were hypertensives. Of the hypertensives, only 39% are on medication. Fewer calories were consumed by the hypertensives. Further, 82.4% of the obese subjects were hypertensives as compared to 52.4% of the non-obese subjects (P<0.001). Only 56 subjects (27.6%) were both diabetic and hypertensive. One third of the elderly had some abnormality in ECG revealing associated coronary heart disease. Forty subjects (11.05%) had complications such as MI and stroke. The prevalence of complications was slightly higher (11.9%) in the hypertensives as compared to the non-hypertensives (9.9%). Thus health education campaigns should be started to increase awareness regarding the risk factors and bring about changes in lifestyle for the control of hypertension and reduction of its complications. PMID- 11896637 TI - Decay of maternally derived measles antibody in central Turkey. AB - The aim of this cross-sectional study was to investigate maternal antibodies in sera of infants below vaccination age and their relation to the immunity status of the mothers. The study group consisted of 184 mothers and their babies aged 0 9 months. Mothers were interviewed to obtain demographic information. Samples of sera were taken from mothers and their babies and tested for measles IgG antibodies. In our study, 174 mothers (94.6%) were immune to measles. Only 78.4% of the 0 month-old infants of seropositive mothers and 26.0% of the 4-9 month-old infants were seropositive. When correlation analysis for antibody titres was made between the seropositive mothers and their seropositive infants, positive correlation was found. It was found that the time for which infants were protected was related to maternal IgG antibody titre. It will be proper to determine the vaccination strategies considering the changing epidemiological characteristics. PMID- 11896638 TI - Beliefs and attitudes of caregivers toward compliance with childhood immunisations in Cameroon. AB - This study analysed factors associated with caregiver compliance and childhood immunisation schedules using a framework based on the Health Belief Model and a cross-sectional questionnaire survey. Participants were caregivers who were enrolling their children (average age 5 y) into class one at randomly selected primary schools in Bamenda, Cameroon. Schools were selected using a stratified random sampling methodology. Seventy-two percent of the 550 participants responded that their children were up-to-date with their immunisations. Perceived susceptibility with an odds ratio (OR)=0.75, perceived severity (OR=0.74), and self-efficacy (OR=1.57) were found to be associated with caregiver compliance to childhood immunisations. Higher level of education and living in an urban location were also found to be associated with increased likelihood of caregivers being up-to-date with their children's immunisations. Results suggest that health service planning should include health education and health promotion programs targeting caregiver compliance with recommended immunisation schedules with resultant improvements in communicable disease control and child health in Cameroon. PMID- 11896641 TI - Dissociating prelexical and postlexical processing of affective information in the two hemispheres: effects of the stimulus presentation format. AB - Using a lexical decision task, the authors investigated whether brain asymmetries in the detection of emotionally negative semantic associations arise only at a perceptually discriminative stage at which lexical analysis is accurate or can already be found at crude and incomplete levels of perceptual representation at which word-nonword discrimination is based solely on guessing. Emotionally negative and neutral items were presented near perceptual threshold in the left and right visual hemifields. Word-nonword discrimination performance as well as the bias to classify a stimulus as a "word" (whether or not it actually is a word) were assessed for a normal, horizontal stimulus presentation format (Experiment 1) and for an unusual, vertical presentation format (Experiment 2). Results show that while the two hemispheres are equally able to detect affective semantic associations at a prelexical processing stage (both experiments), the right hemisphere is superior at a postlexical, perceptually discriminative stage (Experiment 2). Moreover, the findings suggest that only an unusual, nonoverlearned stimulus presentation format allows adequate assessment of the right hemisphere's lexical-semantic skills. PMID- 11896643 TI - Sentence processing strategies in healthy seniors with poor comprehension: an fMRI study. AB - We used fMRI to examine patterns of brain recruitment in 22 healthy seniors, half of whom had selective comprehension difficulty for grammatically complex sentences. We found significantly reduced recruitment of left posterolateral temporal [Brodmann area (BA) 22/21] and left inferior frontal (BA 44/6) cortex in poor comprehenders compared to the healthy seniors with good sentence comprehension, cortical regions previously associated with language comprehension and verbal working memory, respectively. The poor comprehenders demonstrated increased activation of left prefrontal (BA 9/46), right dorsal inferior frontal (BA 44/6), and left posterior cingulate (BA 31/23) cortices for the grammatically simpler sentences that they understood. We hypothesize that these brain regions support an alternate, nongrammatical strategy for processing complex configurations of symbolic information. Moreover, these observations emphasize the crucial role of the left perisylvian network for grammatically guided sentence processing in subjects with good comprehension. PMID- 11896642 TI - Neural correlates of successful and unsuccessful verbal memory encoding. AB - Recent neuroimaging studies suggest that episodic memory encoding involves a network of neocortical structures which may act interdependently with medial temporal lobe (mTL) structures to promote the formation of durable memories, and that activation in certain structures is modulated according to task performance. Functional magnetic resonance imaging (fMRI) was used to determine the neural structures recruited during a verbal episodic encoding task and to examine the relationship between activation during encoding and subsequent recognition memory performance across subjects. Our results show performance-correlated activation during encoding both in neocortical and medial temporal structures. Neocortical activations associated with later successful and unsuccessful recognition memory were found to differ not only in magnitude, but also in hemispheric laterality. These performance-related hemispheric effects, which have not been previously reported, may correspond to between-subject differences in encoding strategy. PMID- 11896644 TI - Tense and syntactic processes in agrammatic speech. AB - This study focuses on inflectional errors in a group of eight, English-speaking people with agrammatism. Subjects were required to provide inflected verbs in declarative sentences and construct negative sentences. The errors made by these speakers are considered in terms of whether they resemble the errors made by children at the Optional Infinitive Stage. Results confirm that the errors differ from those made by children and that a different explanation is required. Explanations offered by Borer and Rohrbacher (1997) and by Friedmann and Grodzinsky (2000) are discussed but are considered inadequate to deal with our data. It is proposed that agrammatic speakers have problems with the implementation of grammar and particularly with syntactic processes such as feature-checking. PMID- 11896645 TI - Competition, inhibition, and semantic judgment errors in Parkinson's disease. AB - Semantic processing errors are symptoms of an up-regulation (schizophrenia) or degradation (Parkinsonism) of dopaminergic pathways. A recent connectionist model attributed errors in the schizophrenic processing of context to increased gain in competitive neural processes. This study extends this "gain hypothesis" by comparing the sensitivity to reduced gain of a simulation of semantic route activation to characteristic semantic judgment errors made by Parkinson's patients in an open search task. Under normal gain conditions, the dominant sense of polysemous words "wins" through competition and lateral inhibition at the word sense level (beta(inh)). For words with very different sense frequencies, decreasing gain by increasing beta(inh) resulted in the dominant word sense winning; however, for words with similar sense frequencies, increasing beta(inh) resulted in the dominant word sense winning only for low to moderate values. At high levels, no clear winner emerged after 200 epochs, with the least dominant sense reaching the maximum activation value. These results are discussed in the context of the Yerkes-Dodson Law, which may provide a theoretical basis for understanding normal and impaired semantic performance in catecholaminergic disorders. PMID- 11896646 TI - Crossmodal temporal order and processing acuity in developmentally dyslexic young adults. AB - We investigated crossmodal temporal performance in processing rapid sequential nonlinguistic events in developmentally dyslexic young adults (ages 20-36 years) and an age- and IQ-matched control group in audiotactile, visuotactile, and audiovisual combinations. Two methods were used for estimating 84% correct temporal acuity thresholds: temporal order judgment (TOJ) and temporal processing acuity (TPA). TPA requires phase difference detection: the judgment of simultaneity/nonsimultaneity of brief stimuli in two parallel, spatially separate triplets. The dyslexic readers' average temporal performance was somewhat poorer in all six comparisons; in audiovisual comparisons the group differences were not statistically significant, however. A principal component analysis indicated that temporal acuity and phonological awareness are related in dyslexic readers. The impairment of temporal input processing seems to be a general correlative feature of dyslexia in children and adults, but the overlap in performance between dyslexic and normal readers suggests that it is not a sufficient reason for developmental reading difficulties. PMID- 11896647 TI - Lateral asymmetry in phonological processing: relating behavioral measures to neuroimaged structures. AB - Recent neuroimaging studies have revealed neuroanatomical centers of activation that appear to be linked specifically to phonological processes. Furthermore, there is evidence that these centers of activation are more likely to be bilaterally represented in women than in men. However, behavioral evidence of such hemispheric asymmetries or of related sex differences is somewhat inconsistent. Two experiments were carried out to determine whether behavioral correlates of the neuroimaging findings could be demonstrated. The first experiment employed a rhyme-matching task used in a recent neuroimaging study, in which two nonsense words were projected successively to the left or right sides of the visual field. The results indicated that the left hemisphere was superior for this task. An analysis of gender effects, however, revealed that the asymmetry was restricted to men. The second experiment employed a lateralized pseudohomophone detection task that was designed to determine whether the results of the first experiment could be confirmed in the context of a phonological task with a simpler structure. This experiment broadly supported the findings of the first experiment. The results support the specialization of the left hemisphere for phonological processing, and the existence of gender differences with respect to these processes. PMID- 11896648 TI - The organization of letter-form representations in written spelling: evidence from acquired dysgraphia. AB - We report on an Italian brain-damaged patient with impaired written spelling. The patient's errors, in different fonts and scripts, consist mainly of letter substitutions (e.g., filo [thread] --> TILO). The results of various tests indicate that letter substitution errors arise because of a deficit in accessing the letter-form representations supporting written spelling. Letter substitutions occurred predominantly between letters with common strokes (e.g., C and G; b and p). Similarities in terms of global letter shape or letter sound were not valid predictors of letter substitution errors. Letter frequency, consonant-vowel status, and letter gemination were factors affecting letter substitution errors. The results of our investigation suggest that information about letter strokes are stored at the level of letter-form representations, and that access to these representations is sensitive to letter frequency. The results further indicate that letter-form representations do not specify whether a letter is a consonant or a vowel, or is a geminate. PMID- 11896649 TI - Semantic representations of word meanings by the cerebral hemispheres. AB - Two priming experiments investigated kind and strength of semantic knowledge underlying known, frontier, and unknown low frequency words. Results from Experiment 1 suggest that known words reflect categorical knowledge, but frontier and unknown words reflect thematic knowledge. Thematic knowledge for frontier words appears to be stronger than that for unknown words. Experiment 2 entailed visual half-field presentation of targets. All facilitory effects were restricted to the lvf/RH, and inhibitory effects to the rvf/LH. Experiment 1 findings were mirrored by the RH. Thematic knowledge appears to precede categorical knowledge for the RH, but the opposite may be true of the LH. Results are also discussed in terms of the RH role in meaning acquisition and metacontrol. PMID- 11896650 TI - Reproducibility of fMRI-determined language lateralization in individual subjects. AB - This study investigated within-subject test-retest reproducibility (i.e., reliability) of language lateralization obtained with fMRI. Nine healthy subjects performed the same set of three different language tasks during two fMRI sessions on separate days (verb generation, antonym generation, and picture naming). A fourth task analysis was added in which the three tasks were analyzed conjointly (combined task analysis, CTA). The CTA targets brain areas that are commonly used in different language tasks, aiming more selectively at language-critical structures. The number of active voxels (i.e., robustness) and calculated lateralization index (LI) were compared across sessions, tasks, subjects, and two a priori defined volumes of interest (classical language regions versus whole hemisphere) for a wide range of statistical thresholds. Robustness and reliability strongly varied between task analyses. The CTA was a robust detector of language-related brain activity, in contrast to the single task approaches. The CTA and verb generation task allowed for reliable calculation of the LI. Higher thresholds yielded a clear increase in left lateralization, which was largest when calculated from active voxels in classical language regions. PMID- 11896651 TI - Effects of sentential context on the processing of unambiguous words by the two cerebral hemispheres. AB - The effect of sentence context on the processing of different aspects of meaning of unambiguous nouns by the two cerebral hemispheres was examined. Participants performed a lexical decision task on target words following two primes, an unambiguous noun preceded by an incomplete sentence. Priming sentences were consistent with either the dominant or the subordinate aspect of meaning of their final unambiguous word. Short and long SOAs were used. A principal finding of this study was that, when compared to unrelated aspects of meaning, for both the short and the long SOAs, the dominant and subordinate aspects of meaning of the unambiguous words were activated regardless of context in both hemispheres. However, the activation of the subordinate aspect of meaning of unambiguous words appears to be more sensitive to sentential context, especially when the unambiguous word is being processed by the left hemisphere. PMID- 11896652 TI - Effect of speech task on intelligibility in dysarthria: a case study of Parkinson's disease. AB - This study assessed intelligibility in a dysarthric patient with Parkinson's disease (PD) across five speech production tasks: spontaneous speech, repetition, reading, repeated singing, and spontaneous singing, using the same phrases for all but spontaneous singing. The results show that this speaker was significantly less intelligible when speaking spontaneously than in the other tasks. Acoustic analysis suggested that relative intensity and word duration were not independently linked to intelligibility, but dysfluencies (from perceptual analysis) and articulatory/resonance patterns (from acoustic records) were related to intelligibility in predictable ways. These data indicate that speech production task may be an important variable to consider during the evaluation of dysarthria. As speech production efficiency was found to vary with task in a patient with Parkinson's disease, these results can be related to recent models of basal ganglia function in motor performance. PMID- 11896653 TI - Recency effects for meaning and form in word selection. AB - Three experiments examined the contribution of phonological availability in selecting words as predicted by interactive activation models of word production. Homophonous words such as week and weak permitted a word's phonological form to be activated on priming trials without selection of its meaning or lemma. Recent production of a homophone failed to significantly increase production of its twin as a sentence completion. However, speakers were significantly more likely to complete a sentence with a recently read or generated unambiguous word. This increase in response probability was unaffected by word frequency. The results constrain the degree to which experience and phonological availability may affect word selection in spoken language production. PMID- 11896654 TI - Novel metaphors appear anomalous at least momentarily: evidence from N400. AB - This study addresses a central question in perception of novel figurative language: whether it is interpreted intelligently and figuratively immediately, or only after a literal interpretation fails. Eighty sentence frames that could plausibly end with a literal, truly anomalous, or figurative word were created. After validation for meaningfulness and figurativeness, the 240 sentences were presented to 11 subjects for event related potential (ERP) recording. ERP's first 200 ms is believed to reflect the structuring of the input; the prominence of a dip at around 400 ms (N400) is said to relate inversely to how expected a word is. Results showed no difference between anomalous and metaphoric ERPs in the early window, metaphoric and literal ERPs converging 300-500 ms after the ending, and significant N400s only for anomalous endings. A follow-up study showed that the metaphoric endings were less frequent (in standardized word norms) than were the anomalous and literal endings and that there were significant differences in cloze probabilities (determined from 24 new subjects) among the three ending types: literal > metaphoric > anomalous. It is possible that the low frequency of the metaphoric element and lower cloze probability of the anomalous one contributed to the processes reflected in the early window, while the incongruity and near-zero cloze probability of the anomalous endings produced an N400 effect in them alone. The structure or parse derived for metaphor during the early window appears to yield a preliminary interpretation suggesting anomaly, while semantic analysis reflected in the later window renders a plausible figurative interpretation. PMID- 11896655 TI - Effects of right and left hemisphere damage on performance of the "Right Hemisphere Communication Battery". AB - A Hebrew adaptation of Gardner and Brownell's (1986) "Right Hemisphere Communication Battery" (HRHCB) was administered to 27 right brain-damaged (RBD) patients, 31 left brain-damaged (LBD) patients, and 21 age-matched normal controls. Both patient groups showed deficits relative to controls and overall there was no difference between the two patient groups. A factor analysis of patients' scores on the HRHCB yielded two interpretable factors, a verbal and a nonverbal one. These factors were not lateralized. Performance of patients on the HRHCB correlated significantly and positively with performance on most tests of basic language functions, measured with a Hebrew adaptation of the "Western Aphasia Battery" (HWAB) and with other cognitive functions measured with standardized neuropsychological tests. There were stronger correlations of HRHCB with subtests of the HWAB in LBD patients and with nonlanguage cognitive tests in RBD patients. In the LBD group, HRHCB subtests' scores correlated negatively with lesion extent in frontal and temporal perisylvian regions. Such localization was not observed in RBD patients. The results argue against selective right hemisphere (RH) involvement in the RHCB, alleged to measure pragmatic aspects of language use, and show, instead, bilateral involvement. The results also argue against a modular organization of these functions of language use, especially in the RH. PMID- 11896656 TI - The relationship between prosody and breathing in spontaneous discourse. AB - This article is concerned with the role of prosody in discourse. Three experiments explored the relationship between inspiration, declination, and syntactic boundaries in normal and RHD participants. Fundamental frequency and intensity were measured at the beginning and end of breath units excised from conversational samples. The results revealed evidence of declination of intensity in all samples measured. However, resetting of fundamental frequency was observed only in the samples of normal participants and then only when a breath coincided with the beginning of a sentence. The results suggest that resetting and declination play separate roles in discourse parsing. PMID- 11896657 TI - Task-related factors in oral motor control: speech and oral diadochokinesis in dysarthria and apraxia of speech. AB - This study was focused on the potential influence of task-related factors on oral motor performance in patients with speech disorders. Sentence production was compared with a nonspeech oral motor task, i.e., oral diadochokinesis. Perceptual and acoustic measures of speech impairment were used as dependent variables. Between-task comparisons were made for subsamples of a population of 140 patients with different motor speech syndromes, including apraxia of speech and cerebellar dysarthria. In a further analysis subgroups were matched for speaking rate. Overall, dysdiadochokinesis was correlated with the degree of speech impairment, but there was a strong interaction between task type and motor speech syndrome. In particular, cerebellar pathology affected DDK to a relatively greater extent than sentence production, while apraxic pathology spared the ability of repeating syllables at maximum speed. PMID- 11896658 TI - Temporal processing and phonological impairment in dyslexia: effect of phoneme lengthening on order judgment of two consonants. AB - The evidence of supporting phonological deficit as a cause of developmental dyslexia has been accumulating rapidly over the past 2 decades, yet the exact mechanisms underlying this deficit remain controversial. Some authors assume that a temporal processing deficit is the source of the phonological disorder observed in dyslexic children. Others maintain that the phonological deficit in dyslexia is basically linguistic, not acoustic, in nature. Three experiments were conducted and tested the impact of the temporal alteration and the impact of complex syllabic structure on consonant order judgments. Thirteen phonological dyslexics (age 10-13) and 10 controls matched for chronologial age were compared on a Temporal Order Judgment (TOJ) task using the succession of two consonants (/p/ /s/) within a cluster. In order to test the possible relevance of the temporal deficit hypothesis, the task also included two additional conditions where either the two stimuli were artificially slowed or two phonological structures were opposed (CCV and CVCV). As expected, the TOJ performance was significantly poorer in dyslexics than in controls. Moreover, in the "slowed speech" condition dyslexics' performance improved to reach the normal controls' level, whereas manipulating the phonological structure complexity provided no significant improvement. Finally dyslexics' performances, especially on the slowed condition, were found correlated with several tests of phonological processing. These results lend support to the general temporal deficit theory of dyslexia. PMID- 11896659 TI - Neuromotor speech deficits in children and adults with spina bifida and hydrocephalus. AB - Acquired cerebellar lesions are associated with motor speech deficits. Spina bifida with hydrocephalus (SBH) is a neurodevelopmental disorder that involves significant dysmorphology of the cerebellum. Videotaped narratives produced by 40 children and adults with SBH and their 40 age-matched controls were coded for three motor speech deficits: dysfluency, ataxic dysarthria (articulatory inaccuracy, prosodic excess, and phonatory-prosodic insufficiency) (Brown, Darley, & Aronson, 1970; Darley, Aronson, & Brown, 1969a), and speech rate. Individuals with SBH had more motor speech deficits than controls. Dysfluency was related to an interaction between chronological age and SBH. Speech rate was related independently to chronological age and SBH. Ataxic dysarthria was related to the biology of SBH, and was associated with both physical phenotype (level of spinal cord lesion) and medical history (number of shunt revisions). The data show that developmental as well as acquired lesions of the cerebellum disrupt motor speech, and add to the developmental role of the cerebellum in the automatization of motor skills, including speech. PMID- 11896660 TI - Assessing resource demands during sentence processing in Parkinson's disease. AB - Patients with Parkinson's disease (PD) have sentence comprehension difficulty, but it is unclear whether this is due to a deficit in grammatical processing or to an executive resource limitation. To assess grammatical processing in PD while minimizing task-related demands, PD patients and healthy control subjects performed a word detection procedure that assesses sensitivity to grammatical agreements in sentences in an "on-line" fashion. With this technique, we found that control subjects and PD patients are equally sensitive to grammatical agreement violations in sentences. A traditional, resource-demanding measure of sentence comprehension was also administered to the same PD patients. In comparison to healthy controls, PD patients were significantly impaired in their relative comprehension of sentences containing object-gap subordinate clauses compared to subject-gap subordinate clauses. Performance on several executive resource measures was also impaired in PD, and this correlated with their comprehension performance. Sensitivity to grammatical agreements with the word detection procedure, in the context of sentence comprehension difficulty on a traditional measure, suggests that PD patients' executive resource limitations contribute to their sentence comprehension difficulty. PMID- 11896661 TI - Reading words and pseudowords: an eye movement study of developmental dyslexia. AB - The pattern of eye movements during reading was studied in 12 developmental dyslexics and in 10 age-matched controls. According to standard reading batteries, dyslexics showed marked reading slowness and prevalently used the sublexical procedure in reading. Eye movements were recorded while they read lists of short and long words or pseudowords. In normal readers, saccade amplitude increased with word length without a concomitant change in the number of saccades; in contrast, the number of saccades increased for long pseudowords. In dyslexics, the eye movement pattern was different. The number of saccades depended on stimulus length for both words and pseudowords while saccade amplitude remained small and constant. The sequential scanning shown by dyslexics for both words and pseudowords appears consistent with the cognitive description of the reading disorder which indicates the preferential use of the sublexical print-to-sound correspondence rules. PMID- 11896663 TI - The Belgian PCB/dioxin incident: analysis of the food chain contamination and health risk evaluation. AB - The Belgian PCB incident occurred at the end of January 1999 when a mixture of polychlorinated biphenyls (PCBs) contaminated with dioxins was accidentally added to a stock of recycled fat used in the production of animal feeds. Although signs of poultry poisoning were noticed by February, 1999, the source and the extent of the contamination were discovered only in May 1999, when it appeared that more than 2500 farms could have been supplied with contaminated feeds. This resulted in a major food crisis, which rapidly extended to the whole country and could be resolved only by the implementation of a large PCB/dioxin food monitoring program. Screening for PCB contamination was based on the determination of the seven PCB markers. When PCB concentrations exceeded the tolerance levels of 0.1 (milk), 0.2 (poultry, bovine, and pig meat), or 1 (animal feed) microg/g fat, dioxins (17 PCDD/Fs congeners) were also determined. At the end of December 1999, the database contained the results of more than 55,000 PCB and 500 dioxin analyses. The study of PCB levels and profiles in contaminated feeds delivered to poultry or pig farms confirmed that the Belgian PCB incident was due to a single source of PCB oil introduced into the food chain at the end of January 1999. This PCB oil had a congeners pattern closely matched to a mixture of Aroclor 1260/1254 in the proportion 75/25. The total amount of PCBs added to recycled fats was estimated at 50 kg (sum of the seven markers) or approximately 150 kg total PCBs, which corresponds to about 100 liters of PCB oil. This PCB mixture contained about 1g TEQ dioxins (more than 90- contributed by PCDFs) and about 2g TEQ dioxin like PCBs. The proportions of PCB 52 and 101 congeners were fairly constant in animal feeds, excluding the possibility of secondary contamination due to fat recycling from contaminated animals. The highest concentrations of PCBs and dioxins were found in poultry and especially in the reproduction animals (hens and chicks), which showed the classical manifestations of chick edema disease. The pigs were also affected but to a lesser extent and no sign of intoxication was observed. The study of PCB/dioxin patterns and of the PCB:dioxin ratios revealed major differences in the metabolism of these compounds by farm animals. Whereas the PCBs:dioxins ratio was fairly constant in all poultry products with a mean value similar to that found in contaminated feeds (50,000), in pigs this ratio was both much higher and more variable (values up to 10,000,000), reflecting a faster elimination of dioxins than PCBs in these animals. These metabolic differences also emerged from the PCB and dioxin patterns which were altered much more in pigs than in poultry. Although the most contaminated food products (chicken meat) had PCB and dioxin levels more than 100 times above maximal recommended values, it is unlikely that this incident could have caused adverse effects in the general population of Belgium. A doubling of the PCB and dioxin burden of the young adult population would require the consumption of, respectively, 10 and 20 highly contaminated meals. In view of the very limited proportion of the poultry chain effectively contaminated during the incident (around 2%), such an extreme scenario was quite improbable for the general population except perhaps for farmers consuming their own products. But even in that case, it would have meant going back to the levels in the 1980s or attaining the body burden of subjects regularly eating contaminated seafood. PMID- 11896664 TI - Brief butyltin exposure induces irreversible inhibition of the cytotoxic function on human natural killer cells, in vitro. AB - Despite mounting evidence on butyltin (BT) contamination and related immunotoxic effects on wildlife, very little is known about BT-associated immunotoxic effects on humans, particularly the effects on human natural killer (NK) lymphocyte function. Our earlier studies demonstrated that in vitro exposure to environmentally relevant concentrations of BTs negatively affect human NK cells and that there are measurable levels of BTs in human blood. In this study we examined whether the inhibition of NK cell cytotoxic function induced by a brief exposure (1 h) to BTs is reversible when the cells are allowed to recover in BT free media for up to 6 days. Standard methods were used in chemical preparation, blood sampling, NK cell isolation, and 51-Chromium release assay. The results revealed that exposure to 300 nM TBT for 1 h caused an approximately 65- decrease in NK cytotoxic function, whether the lymphocytes were given as long as a 6-day recovery period or no recovery period. There was no recovery (nor any further loss) of NK cytotoxic function following removal of the compound. Exposure to 5 microM DBT for 1 h showed a 41% decrease in cytotoxic function with 0-h recovery and an 83% decrease after a 24-h recovery period. Thus, not only is there no significant recovery of NK cytotoxic function when the lymphocytes are allowed to incubate in BT-free medium for up to 6 days but there is additional loss of cytotoxic function. The results indicated that short-term exposure to BTs causes persistent negative effects on NK cell ability to kill cancer cells. PMID- 11896665 TI - Effect of cadmium on the nucleoli of meristematic cells of onion Allium cepa L: an ultrastructural study. AB - This study of the effect of cadmium on nucleolar ultrastructure was carried out with meristematic cell populations of Allium cepa L. Meristems, grown at 25 degrees C, were treated with 10 ppm cadmium chloride. Conventional and silver staining techniques were carried out, and the ultrastructure was analyzed using electron microscopy. Observation showed alterations in the nucleoli of the cells that had been treated with cadmium and this effect varied according to the time of exposure to the metal. After 4h of treatment, nucleolar segregation was observed in interphase, probably because of the effect of cadmium on the synthesis of ribosomal RNA precursors. A decrease in the fibrillar to granular component ratio also occurred in the cells exposed to Cd2+ for 8 h. Some changes were observed in the G1 cells; their chromatin still remained very condensed, and prenucleolus bodies remained scattered within the nucleus. At the same time, there was a large amount of interchromatin granules. These changes produced by cadmium resembled those produced during inhibition of RNA synthesis. The fibrillar bodies, another morphologic feature, resulting from a blocked transcription, were also evidenced. All these observations suggest that one of the ways that cadmium exercises its toxicity is by altering the biosynthesis of the preribosomal RNA precursor. PMID- 11896667 TI - Mercury exposure among residents of a building block in Shiraz, Iran. AB - Exposure to mercury can cause serious multiorgan damage affecting the central nervous system, kidneys, liver, lungs, spleen, bone marrow, and skin. At the end of the summer of 1999, the accidental leakage of 4 liters of mercury from a container into the waterway canals resulted in mass exposure to elemental mercury among the residents of a building block of a residential area of the city of Shiraz, in the south of Iran. One hundred and eleven individuals who experienced exposure to elemental mercury were investigated. Twenty-four-hour measurement of the urine mercury level-revealed a toxic level of more than 20 microg/L in 6 children and 3 adults (including a pregnant woman). Despite normal physical and laboratory (CBC, renal and liver function tests, and urinalysis) findings, dimercaprol was prescribed. One month later during the course of the follow-up the urine mercury level in 6 patients, including the pregnant woman from the same family, was found to be again at a toxic level. The pregnant mother from the same family aborted her fetus; however, due to the lack of equipment for measuring the serum mercury level, it was not possible to confirm the relation between the mercury toxicity and the abortion. This family had kept mercury in their kitchen against health workers' instructions. The attractive physical and chemical properties of mercury could explain the continuity of exposure and poisoning in these 6 cases. It is concluded that prophylactic therapy in the presence of toxic levels of mercury, despite the presence of an asymptomatic state in exposed residents, is effective in preventing the development of signs and symptoms, though instruction of high-risk cases is the best way to combat it. PMID- 11896666 TI - Chemiluminescent detection of induced reactive oxygen metabolite production of human polymorphonuclear leucocytes by anthophyllite asbestos. AB - Incidences of lung cancer and pleural plaque have been reported in relation to exposure to anthophyllite asbestos. To investigate the pathogenic mechanisms of anthophyllite, chemiluminescence (CL) detection of reactive oxygen metabolite (ROM) generation of human polymorphonuclear leucocytes (PMN) stimulated by anthophyllite asbestos was determined and compared with that of other asbestos and mineral fiber samples. When anthophyllite fiber sample was mixed with the luminol-primed PMN, high levels of CL which exhibited a specific time course characterized by two separate peaks were induced. The CL induced by anthophyllite sample was greater than that induced by chrysotile, crocidolite, and amosite asbestos. We further investigated the two peaks of CL using specific inhibitors of signal transduction mechanisms. The two peaks of CL by anthophyllite sample were different in sensitivity to cytochalasin B and genistein; the former relates to the cytoskeleton-dependent mechanism and the latter has been shown to inhibit tyrosine kinase, which resides in the pathway to cause PMN activation. The strong ROM reaction of PMN by anthophyllite suggests that the surface characteristics of the fiber may participate in the pathogenic mechanisms of anthophyllite asbestos. PMID- 11896668 TI - In-vehicle exposure to aldehydes while commuting on real commuter routes in a Korean urban area. AB - This study evaluated in-vehicle exposure to formaldehyde and acetaldehyde on actual commuting routes, while focusing on three factors (transportation mode, passenger car type, and commuting season). A total of 40 passenger car commuters and 20 public bus commuters were recruited. The same commuters participated in both the summer and the winter studies. The transportation mode and passenger car type were found to have little effect on the in-vehicle aldehyde levels. Conversely, the commuting season did influence the in-vehicle aldehyde levels. Meanwhile, the mean formaldehyde-to-acetaldehyde concentration ratios were similar in both the passenger cars and the public buses, plus there were significant correlations (P<0.0001) between formaldehyde and acetaldehyde concentrations for both the passenger cars and the public buses. This study also confirmed that, under Korean commuting conditions, vehicle interiors are an important microenvironment for exposure to formaldehyde and acetaldehyde. The mean in-car concentrations were 20.0 and 8.9 ppb for formaldehyde and acetaldehyde, respectively. Similarly, the mean in-bus concentrations were 21.2 and 9.1 ppb for formaldehyde and acetaldehyde, respectively. Furthermore, the in vehicle formaldehyde levels were higher than those of a previous California study. PMID- 11896669 TI - Gill cellular changes induced by copper exposure in the South American tropical freshwater fish Prochilodus scrofa. AB - The cellular changes in gill tissue induced by exposure to copper were studied in the tropical freshwater fish Prochilodus scrofa, with emphasis on chloride and pavement cells. Damage to gills included epithelial changes such as lifting, rupture, peeling of lamellar epithelium, lamellar fusion, hyperplasia, and cellular hypertrophy. Cell degeneration by necrosis and apoptosis was intense in fish exposed to 25 and 29 microg Cu L-1. Pavement cells showed microridge reduction on their surface. Chloride cells proliferated in the lamellar epithelia close to the onset of the lamellae. However, no changes in total chloride cell density in contact with the water were observed. The chloride cell apical area of fish exposed to copper increased, but only fish exposed to 25 microg Cu L-1 showed significant increase in the chloride cell fractional area. At this water copper concentration, almost 60% of the chloride cells were apoptotic. Necrotic chloride cells increased with copper in water, reaching 70% in fish exposed to 29 microg Cu L-1 (=LC50 calculated for this species). Pavement and chloride cell proliferation and hypertrophy on lamellar epithelia increased the thickness of the water-blood barrier. Our findings suggest severe impairment of ion regulation and gas transfer of fish exposed to copper. PMID- 11896670 TI - Soil contamination detected using bacterial and plant mutagenicity tests and chemical analyses. AB - Soil contaminants are common in industrialized countries, causing widespread contamination directly of soil and indirectly of ground water and food. Among these pollutants particular attention should be paid to soil mutagens and carcinogens due to their potentially hazardous effects on animal populations and human health. The aim of this research was to evaluate the genotoxicity of contaminated soils by means of an integrated chemical/biological approach, using a short-term bacterial mutagenicity test (Ames test), a plant genotoxicity test (Tradescantia/micronucleus test), and chemical analyses. Soil samples were collected in a highly industrialized area in the Lombardy region, in Northern Italy. Soil samples were extracted with water or with organic solvents. Water extracts of soil samples were tested using the Tradescantia genotoxicity test and organic solvent extracts were analyzed for their polycyclic aromatic hydrocarbon (PAH) concentrations and for their mutagenicity with the Ames test. Heavy metal concentrations were also studied. Some soil samples showed mutagenic activity with the Ames test and clastogenicity with the Tradescantia/micronucleus test. The same soils showed high concentrations of genotoxic PAH and heavy metals. PMID- 11896671 TI - Organochlorine exposures and breast cancer risk in New York City women. PMID- 11896674 TI - Biotransformation reactions of five-membered aromatic heterocyclic rings. PMID- 11896675 TI - 4,5-Epoxy-2(E)-decenal-induced formation of 1,N(6)-etheno-2'-deoxyadenosine and 1,N(2)-etheno-2'-deoxyguanosine adducts. AB - trans-4,5-Epoxy-2(E)-decenal reacted with 2'-deoxyadenosine to give 1,N(6)-etheno 2'-deoxyadenosine as well as other 2'-deoxyadenosine adducts. It also reacted with 2'-deoxyguanosine to give 1,N(2)-etheno-2'-deoxyguanosine and other 2' deoxyguanosine adducts. Synthetic trans-4,5-epoxy-2(E)-decenal was quite stable under the reaction conditions that were used. It was not contaminated with 2,3 epoxyoctanal, a potential precursor to the formation of unsubstituted etheno adducts. Furthermore, using a sensitive LC/MS assay, it was possible to show that no 2,3-epoxyoctanal was formed during prolonged incubations of trans-4,5-epoxy 2(E)-decenal. Therefore, trans-4,5-epoxy-2(E)-decenal, a primary product of lipid peroxidation, is a precursor to the formation of 1,N(6)-etheno-2'-deoxyadenosine and 1,N(2)-etheno-2'-deoxyguanosine. There is no need for an additional oxidation step such as would be required if trans,trans-2,4-decadienal or 4-hydroxy-2 nonenal were the lipid hydroperoxide decomposition products that initiated the formation of unsubstituted etheno adducts. These findings provide an important link between a primary product of lipid peroxidation and a mutagenic DNA lesion that has been detected in human tissues. PMID- 11896676 TI - 32P-Postlabeling/polyacrylamide gel electrophoresis analysis: application to the detection of DNA adducts. AB - 32P-Postlabeling analysis is a powerful technique to detect DNA adducts. Polyethylenimine-cellulose TLC plates are generally used to separate (32)P labeled adducts using several different buffers. However, separation by TLC is time-consuming and labor-intensive for a large number of DNA samples. To expedite analyses, nondenaturing 30% polyacrylamide gel electrophoresis (PAGE) has been adapted for the (32)P-postlabeling analysis. The major advantages of this technique are as follows: (a) many DNA samples can be loaded concomitantly on the PAGE with standard markers; (b) DNA adducts can be resolved in only a few hours; and (c) exposure to (32)P during handing can be minimized. To show the usefulness of (32)P-postlabeling/PAGE analysis, the formation of a tamoxifen (TAM)-DNA adduct resulting from O-sulfonation of alpha-hydroxytamoxifen was demonstrated. In addition, to quantify TAM adducts, oligodeoxynucleotides containing diastereoisomers of alpha-(N(2)-deoxyguanosinyl)tamoxifen can be used as standards. The detection limit of this assay for 5 microg of DNA was approximately 7 adducts/10(9) nucleotides. The (32)P-postlabeling/PAGE analysis can also be used to detect DNA adducts derived from benzo[a]pyrene diol epoxide, 2-acetylaminofluorene, and 4-hydroxyequilenin. PMID- 11896677 TI - Reaction of malondialdehyde-DNA adducts with hydrazines-development of a facile assay for quantification of malondialdehyde equivalents in DNA. AB - Malondialdehyde is a ubiquitous product of lipid peroxidation that reacts with DNA to form premutagenic lesions. Principal among them is pyrimido-[1,2 alpha]purin-10(3H)-one (M(1)G). M(1)G has recently been found to be a reactive electrophile in DNA that couples with amines at basic pH or hydroxylamines at neutral pH. We explored the reaction of M(1)G with hydrazines because of the possibility that the latter could act as bifunctional nucleophiles to strip the malondialdehyde equivalent from DNA. Pentafluorophenylhydrazine reacted rapidly with M(1)G to form a hydrazone conjugate. This hydrazone was stable at room temperature and did not cyclize to form the corresponding pyrazole. In contrast, phenylhydrazine and benzylhydrazine reacted with M(1)G to form phenylpyrazole and benzylpyrazole, respectively. Pentafluorobenzylhydrazine reacted rapidly with M(1)G to form pentafluorobenzylpyrazole and dG in near quantitative yield. This reaction formed the basis for a quantitative assay for the presence of M(1)G or M(1)G equivalents in DNA or protein that utilized gas chromatography/negative chemical ionization mass spectrometry. The assay was extended to the oxopropenyl donors, M(1)A, base propenal, and N(epsilon)-3-oxopropenyl-lysine. Analysis of DNA treated with bleomycin demonstrated a linear increase in the level of oxopropenyl groups that plateaued at approximately 1 oxopropenyl group/100 bases at a bleomycin concentration of 200 microM. Parallel analysis of M(1)G in the samples revealed that this adduct represents a small fraction of the total oxopropenyl units generated in DNA by treatment with bleomycin. PMID- 11896678 TI - In vitro inhibition of the enzymatic activity of tumor suppressor FHIT gene product by carcinogenic transition metals. AB - FHIT (Fragile Histidine Triad) is a human tumor suppressor gene. The Fhit protein is believed to inhibit tumor growth by inducing apoptosis through interaction with diadenosine triphosphate (Ap(3)A). The latter is first sequestered and eventually hydrolyzed by Fhit to ADP and AMP. Thus, the balance between the cellular Ap(3)A level and Fhit enzymatic activity may affect cell death or survival. Increasing the Ap(3)A level, e.g., by inhibition of the enzyme, should prevent apoptosis and thus sustain tumorigenesis. To test if certain carcinogenic transition metals could inhibit the enzymatic activity of Fhit, purified human Fhit protein [30 nM in 1.25 mM poly(vinylpyrrolidone)], expressed in and isolated from E. coli, was incubated at pH 6.8 (50 mM HEPES buffer in 150 mM NaCl) with 120 microM Ap(3)A in the presence of 5 mM Mg(II) (activating cation) and 0-100 microM Ni(II), Cu(II), Zn(II), Cd(II), Co(II), Cr(III), As(III), or As(V). The reaction mixtures were analyzed by HPLC. The results revealed a strong inhibitory potential of Cu(II) [0.4], followed by Ni(II) [3.5] >or= Zn(II) [7.0] >> Cr(III) [73] > Cd(II) [98] >> Co(II) [432] [the numbers in brackets are IC(50) values, microM]. As(III) and As(V) had no effect. As revealed by spectrophotometry, mass spectrometry, and gel electrophoresis, the exceptionally strong inhibition by Cu(II) was associated with Fhit dimerization through formation of a disulfide bond. The other metals and also H(2)O(2) and NO did not cause the dimerization. Thus, the effect of Cu(II) must be due to its reaction with Cys-39 bearing the only thiol group in Fhit monomer. Since Cys-39 is not readily accessible in the Fhit molecule, the reaction is most likely facilitated by conformational changes which follow the coordination of Cu(II) by the surface histidines 35, 94, and/or 96. The observed inhibition of Fhit may be mechanistically involved in metal mediated toxicity and carcinogenesis. PMID- 11896679 TI - Pesticide Roundup provokes cell division dysfunction at the level of CDK1/cyclin B activation. AB - To assess human health risk from environmental chemicals, we have studied the effect on cell cycle regulation of the widely used glyphosate-containing pesticide Roundup. As a model system we have used sea urchin embryonic first divisions following fertilization, which are appropriate for the study of universal cell cycle regulation without interference with transcription. We show that 0.8% Roundup (containing 8 mM glyphosate) induces a delay in the kinetic of the first cell cleavage of sea urchin embryos. The delay is dependent on the concentration of Roundup. The delay in the cell cycle could be induced using increasing glyphosate concentrations (1-10 mM) in the presence of a subthreshold concentration of Roundup 0.2%, while glyphosate alone was ineffective, thus indicating synergy between glyphosate and Roundup formulation products. The effect of Roundup was not lethal and involved a delay in entry into M-phase of the cell cycle, as judged cytologically. Since CDK1/cyclin B regulates universally the M-phase of the cell cycle, we analyzed CDK1/cyclin B activation during the first division of early development. Roundup delayed the activation of CDK1/cyclin B in vivo. Roundup inhibited also the global protein synthetic rate without preventing the accumulation of cyclin B. In summary, Roundup affects cell cycle regulation by delaying activation of the CDK1/cyclin B complex, by synergic effect of glyphosate and formulation products. Considering the universality among species of the CDK1/cyclin B regulator, our results question the safety of glyphosate and Roundup on human health. PMID- 11896680 TI - Microsomal activation of dibenzo[def,mno]chrysene (anthanthrene), a hexacyclic aromatic hydrocarbon without a bay-region, to mutagenic metabolites. AB - Metabolically formed dihydrodiol epoxides in the bay-region of polycyclic aromatic hydrocarbons are thought to be responsible for the genotoxic properties of these environmental pollutants. The hexacyclic aromatic hydrocarbon dibenzo[def,mno]chrysene (anthanthrene), although lacking this structural feature, was found to exhibit considerable bacterial mutagenicity in histidine dependent strains TA97, TA98, TA100, and TA104 of S. typhimurium in the range of 18-40 his(+)-revertant colonies/nmol after metabolic activation with the hepatic postmitochondrial fraction of Sprague-Dawley rats treated with Aroclor 1254. This mutagenic effect amounted to 44-84% of the values determined with benzo[a]pyrene under the same conditions. The specific mutagenicity of anthanthrene in strain TA100 obtained with the cell fraction of untreated animals was 6 his(+)-revertant colonies/nmol and increased 2.7-fold after treatment with phenobarbital and 4.5 fold after treatment with 3-methylcholanthrene. To elucidate the metabolic pathways leading to genotoxic metabolites, the microsomal biotransformation of anthanthrene was investigated. A combination of chromatographic, spectroscopic, and biochemical methods allowed the identification of the trans-4,5-dihydrodiol, 4,5-oxide, 4,5-, 1,6-, 3,6-, and 6,12-quinones, and 1- and 3-phenols. Furthermore, two diphenols derived from the 3-phenol, possibly the 3,6 and 3,9 positional isomers, as well as two phenol dihydrodiols were isolated. Three pathways of microsomal biotransformation of anthanthrene could be distinguished: The K-region metabolites are formed via pathway I dominated by monooxygenases of the P450 1B subfamily. On pathway II the polynuclear quinones of anthanthrene are formed. Pathway III is preferentially catalyzed by monooxygenases of the P450 1A subfamily and leads to the mono- and diphenols of anthanthrene. The K-region oxide and the 3-phenol are the only metabolites of anthanthrene with strong intrinsic mutagenicity, qualifying them as ultimate mutagens or their precursors. From the intrinsic mutagenicity of these two metabolites and their metabolic formation, the maximal mutagenic effect was calculated. This demonstrates the dominating role of pathway III in the mutagenicity of anthanthrene under conditions where it exhibits the strongest bacterial mutagenicity. PMID- 11896681 TI - The regioselectivity of glutathione adduct formation with flavonoid quinone/quinone methides is pH-dependent. AB - In the present study, the formation of glutathionyl adducts from a series of 3',4'-dihydroxy flavonoid o-quinone/p-quinone methides was investigated with special emphasis on the regioselectivity of the glutathione addition as a function of pH. The flavonoid o-quinones were generated using horseradish peroxidase, and upon purification by HPLC, the glutathionyl adducts were identified by LC/MS as well as (1)H and (13)C NMR. The major pH effect observed for the glutathione conjugation of taxifolin and luteolin quinone is on the rate of taxifolin and luteolin conversion and, as a result, on the ratio of mono- to diglutathione adduct formation. With fisetin, 3,3',4'-trihydroxyflavone, and quercetin, decreasing the pH results in a pathway in which glutathionyl adduct formation occurs in the C ring of the flavonoid, being initiated by hydration of the quinone and H(2)O adduct formation also in the C ring of the flavonoid. With increasing pH, for fisetin and 3,3',4'-trihydroxyflavone glutathione adduct formation of the quinone occurs in the B ring at C2' as the preferential site. For quercetin, the adduct formation of its quinone/quinone methide shifts from the C ring at pH 3.5, to the A ring at pH 7.0, to the B ring at pH 9.5, indicating a significant influence of the pH and deprotonation state on the chemical electrophilic behavior of quercetin quinone/quinone methide. Together the results of the present study elucidate the mechanism of the pH-dependent electrophilic behavior of B ring catechol flavonoids, which appears more straightforward than previously foreseen. PMID- 11896682 TI - 1,3-Dinitrobenzene metabolism and protein binding. AB - 1,3-Dinitrobenzene is a testicular toxicant, which produces a lesion in the seminiferous tubules of the rat. In the present study, we investigated which subcellular fractions of the seminiferous tubules are capable of 1,3 dinitrobenzene metabolism and protein adduct formation. Subcellular fractions of the liver were used as positive controls and to further investigate potentially important binding proteins. Microsomes, cytosol, and mitochondria prepared from each tissue were incubated with 200 microM [(14)C]1,3-dinitrobenzene and 2 mM NADH or NADPH. Since nitroreduction is an oxygen sensitive metabolic pathway, incubations were carried out in the presence and absence of oxygen. Under anaerobic conditions, 1,3-dinitrobenzene was metabolized to nitroaniline and/or nitrophenylhydroxylamine. Metabolite formation was inhibited under aerobic conditions, suggesting the presence of an oxygen-dependent redox-cycle. For the seminiferous tubules, no metabolites were generated under aerobic conditions. In the absence of oxygen, only the mitochondria produced 1,3-dinitrobenzene metabolites. For the liver, under anaerobic conditions, all three subcellular fractions produced 1,3-dinitrobenzene metabolites with the microsomes containing the greatest activity. However, under aerobic conditions, only the microsomes generated metabolites. One-dimensional gel electrophoresis demonstrated that protein adduct formation within the liver and seminiferous tubule subcellular fractions correlated with metabolite formation. Addition of GSH to seminiferous tubule mitochondrial incubations decreased the amount of (14)C-labeled protein. Moreover, when seminiferous tubule mitochondria were incubated with 1,3 dinitrobenzene at an increased protein concentration, radioactive labeling of a 54 kDa protein became more prominent. Two-dimensional gel electrophoresis of liver mitochondrial protein incubated with [(14)C]1,3-dinitrobenzene and NADPH yielded three predominantly radiolabeled proteins of the same approximate size (54 kDa). Amino acid sequencing identified each of these proteins as rat mitochondrial aldehyde dehydrogenase. PMID- 11896684 TI - Formation of cyclic deoxyguanosine adducts from omega-3 and omega-6 polyunsaturated fatty acids under oxidative conditions. AB - The discovery of the cyclic 1,N(2)-propanodeoxyguanosine adducts of acrolein (Acr), crotonaldehyde (Cro), and t-4-hydroxy-2-nonenal (HNE) as endogenous DNA lesions from lipid peroxidation has raised questions regarding the role of different types of fatty acids as sources for their formation. In this study, we carried out reactions at pH 7 and 37 degrees C with deoxyguanosine 5' monophosphate and omega-3 polyunsaturated fatty acids (PUFAs), including docosahexaenoic acid (DHA), linolenic acid (LNA), and eicosapentaenoic acid (EPA); or omega-6 PUFAs, including linoleic acid (LA) and arachidonic acid (AA), each in the presence of ferrous sulfate. The formation of Acr, Cro, and HNE derived 1,N(2)-propanodeoxyguanosine adducts (Acr-, Cro-, and HNE-dG) in the incubation mixture was determined by reversed-phase HPLC analysis. The results showed that Acr and Cro adducts are primarily derived from omega-3 PUFAs, although Acr adducts are also formed, to a lesser extent, from oxidized AA and LA. HNE-dG adducts were detected exclusively in incubations with AA. The kinetics of the formation of these adducts was determined during incubations for 2 weeks and 5 days. The rate of Acr adduct formation was about 5-10-fold that of Cro adducts, depending on the type of PUFAs, and the rate of formation of HNE adducts from AA was also considerably slower than that of Acr adducts. Unlike other cyclic adducts, the formation of Acr adducts was independent of types of PUFAs, but its yield was proportional to the number of double bonds in the fatty acid. Only one of the isomeric Acr adducts was detected, and its stereoselective formation is consistent with that observed previously in vivo. Two previously unknown cyclic adducts, one derived from pentenal and the other from heptenal, were also detected as products from omega-3 and omega-6 fatty acids, respectively. This study demonstrated the specificity for the formation of the cyclic adducts of Acr, Cro, and HNE and other related enals by oxidation of omega 3 and omega-6 PUFAs. These results may be important for the understanding of the specific roles of different types of fatty acids in tumorigenesis. PMID- 11896683 TI - 1,3-Dinitrobenzene metabolism and GSH depletion. AB - Previous work demonstrated that the mitochondrial fraction of rat seminiferous tubules is capable of metabolizing 1,3-dinitrobenzene, using NADPH as a cofactor. Moreover, 1,3-dinitrobenzene treatment of rat tubules caused a decrease in mitochondrial GSH levels. In situ mitochondrial metabolism of 1,3-dinitrobenzene may have caused this depletion through the production of reactive oxygen intermediates, generating oxidative stress and/or one or more metabolites of 1,3 dinitrobenzene which reacted nonenzymatically with GSH. The goal of this study is to investigate which of these two potential mechanisms may have caused the observed GSH depletion. Liver microsomes, known to rapidly metabolize 1,3 dinitrobenzene, generated the superoxide anion radical when incubated with 1,3 dinitrobenzene and NADPH. However, with the seminiferous tubule mitochondria, no oxygen radicals were detected. Hence, the aforementioned GSH depletion is unlikely due to the production of reactive oxygen intermediates from in situ mitochondrial metabolism of 1,3-dinitrobenzene. To investigate the ability of 1,3 dinitrobenzene metabolites to deplete seminiferous tubule mitochondrial GSH, mitochondria were incubated with 1,3-dinitrobenzene and NADPH. Loss of GSH correlated with the appearance of the 1,3-dinitrobenzene metabolites, nitrophenylhydroxylamine and nitroaniline. Subsequent investigation demonstrated that the metabolites, nitrosonitrobenzene, known to react nonenzymatically with nonprotein sulfhydryls, and nitrophenylhydroxylamine both oxidized seminiferous tubule mitochondrial GSH. Further studies suggested that nitrophenylhydroxylamine could deplete GSH via a free radical mechanism. In aqueous solution, this metabolite was shown to exist in equilibrium with a radical form, thought to be the hydronitroxide radical. The addition of GSH eliminated the signal, implying that the radical reacted nonenzymatically with GSH. In conclusion, the data in this study suggest that the decrease in mitochondrial GSH observed in DNB-treated seminiferous tubules is due to the formation of NPHA and NNB and not reactive oxygen intermediates. PMID- 11896685 TI - Characterization of nucleoside adducts of cis-2-butene-1,4-dial, a reactive metabolite of furan. AB - Furan is a hepatic toxicant and carcinogen in rodents. Its microsomal metabolite, cis-2-butene-1,4-dial, is mutagenic in the Ames assay. Consistent with this observation, cis-2-butene-1,4-dial reacts with 2'-deoxycytidine, 2' deoxyguanosine, and 2'-deoxyadenosine to form diastereomeric adducts. HPLC analysis indicated that the rate of reaction with deoxyribonucleosides was dependent on pH. At pH 6.5, the relative reactivity was 2'-deoxycytidine > 2' deoxyguanosine > 2'-deoxyadenosine whereas it was 2'-deoxyguanosine > 2' deoxycytidine > 2'-deoxyadenosine at pH 8.0. Thymidine did not react with cis-2 butene-1,4-dial. The primary 2'-deoxyguanosine and 2'-deoxyadenosine reaction products were unstable and decomposed to secondary products. NMR and mass spectral analysis indicated that the initial 2'-deoxyadenosine and 2' deoxyguanosine reaction products were hemiacetal forms of 3-(2'-deoxy-beta-D erthyropentafuranosyl)-3,5,6,7-tetrahydro-6-hydroxy-7-(ethane-2''-al)-9H imidazo[1,2-alpha]purine-9-one (structure 2) and 3-(2'-deoxy-beta-D erythropentafuranosyl)-3,6,7,8-tetrahydro-7-(ethane-2''-al)-8-hydroxy-3H imidazo[2,1-i]purine (structure 4), respectively. These adducts resulted from the addition of cis-2-butene-1,4-dial to the exo- and endocyclic nitrogens of 2' deoxyadenosine and 2'-deoxyguanosine. The data provide support for the hypothesis that cis-2-butene-1,4-dial is an important genotoxic intermediate in furan induced carcinogenesis. PMID- 11896686 TI - Modified guanines representing O(6)-alkylation by the cyclophosphamide metabolites acrolein and chloroacetaldehyde: synthesis, stability, and ab initio studies. AB - Alkylation of DNA by acrolein and/or chloroacetaldehyde may result in the mutations that lead to the therapy-induced leukemia associated with cyclophosphamide (and ifosfamide) treatment. O(6)-(n-Propanalyl)guanine (O(6) PAG) and O(6)-(ethanalyl)guanine (O(6)-EAG) were synthesized for use as authentic standards in investigations of DNA alkylation by acrolein and chloroacetaldehyde, respectively. Preparation of the O-methyl oximes of these aldehydes aided in confirming the structural assignments of O(6)-PAG and O(6)-EAG. HPLC was used to study the stability of O(6)-PAG under a variety of conditions. The decomposition of O(6)-PAG was attributed to an alpha,beta-elimination reaction resulting in the formation of guanine and acrolein. In 0.1 M phosphate-DMSO (9:1), O(6)-PAG (1-10 mM) had a half-life of approximately 1 h (pH 7.4, 37 degrees C). In 0.05 M Tris DMSO (9:1), the apparent half-life of O(6)-PAG (1-10 mM) was approximately 16 h (pH 7.4, 37 degrees C). The increased lifetime under the latter conditions was attributed to a reversible reaction between Tris and the aldehydic functionality of O(6)-PAG to give a more stable oxazolidine. Under conditions similar to those that would be used for hydrolysis of DNA [0.1 M HCl-DMSO (98:2), pH 1.3, 70 degrees C, 30 min], there was an estimated 10-35% loss of O(6)-PAG. Under the same conditions, O(6)-EAG had apparent half-lives of 6.6 h (phosphate-DMSO) and 2.5 days (Tris-DMSO) and the estimated loss at pH 1.3 over 30 min (70 degrees C) was 15-20%. Ab initio quantum chemical calculations were used to understand the energy factors that underlie the occurrence of O- versus N-alkylations as well as possible, subsequent intramolecular cyclizations. Simulations of the free energies of reactions between acrolein and guanine indicated that N-alkylation was favored over O(6)()-alkylation and that cyclizations to tautomers were most favorable if they involved the N-1 or NH(2) positions. PMID- 11896687 TI - Delineating novel metabolic pathways of DPC 963, a non-nucleoside reverse transcriptase inhibitor, in rats. Characterization of glutathione conjugates of postulated oxirene and benzoquinone imine intermediates by LC/MS and LC/NMR. AB - The metabolic activation of (S)-5,6-difluoro-4-cyclopropylethynyl-4 trifluoromethyl-3,4-dihydro-2(1H)-quinazolinone, DPC 963, in rats was investigated by identifying and characterizing the GSH and mercapturic acid conjugates excreted in the bile and urine, respectively. The structures of these adducts, which were unequivocally elucidated by LC/MS/MS and NMR experiments, revealed the existence of at least three distinct metabolic pathways leading to these products. One of the pathways, which has been described previously, involves the activation of the acetylene group after an initial hydroxylation on the methine carbon of the cyclopropyl ring. Metabolite M1 was demonstrated to be formed via this pathway after an enzymatic addition of GSH across the triple bond of the substituted acetylene. The second pathway, also previously described, leads to diastereoisomeric GSH adducts M3 and M4 after the formation of a highly reactive oxirene intermediate. This postulated oxirene subsequently rearranges to an alpha, beta-unsaturated cyclobutenyl ketone intermediate capable of undergoing a 1,4-Michael addition with a nucleophile such as GSH. In addition to these pathways, DPC 963 was found to undergo a metabolic activation previously undescribed for structural analogues of this compound. It is postulated that an oxidative defluorination mediated by cytochrome P450 leads to the formation of a putative benzoquinone imine intermediate which subsequently reacts with GSH to form two aromatic ring-substituted regioisomeric conjugates, M5 and M6. In addition to forming the GSH adducts, the benzoquinone imine was also found to be reduced to its unreactive hydroquinone metabolite, which was excreted as the glucuronide conjugate in rat bile. Studies with induced rat microsomes, cDNA expressed rat P450 isozymes, and polyclonal antibodies against rat P450 clearly demonstrated that the rat P450s 3A1/3A2 were responsible for the formation of postulated oxirene and benzoquinone intermediates. PMID- 11896688 TI - UVA light-induced DNA cleavage by isomeric methylbenz[a]anthracenes. AB - UVA light-induced DNA single strand cleavage by a set of 12 monomethyl substituted benz[a]anthracenes (MBAs) along with their parent compound, benz[a]anthracene (BA), and the potent carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA), was studied. On the basis of the relative DNA single strand photocleavage efficiency of the fourteen compounds, they are divided into three groups: (1) strong DNA cleavers, 4-MBA, 5-MBA, 6-MBA, 8-MBA, 9-MBA, 10-MBA, and BA; (2) medium DNA cleavers, 1-MBA, 2-MBA, 3-MBA, and 11-MBA; and (3) weak DNA cleavers, 7-MBA, 12-MBA, and DMBA. The relative DNA photocleavage efficiency parallels very well with the energy gap between the highest-occupied-molecular-orbital (HOMO) and the lowest-unoccupied-molecular-orbital (LUMO) of each MBA, indicating that the DNA cleavage is related to their excited-state properties. The 7 and 12 positions of BA are two unique sites. Methyl substitution at either 7 or 12 (or both) positions lowers the HOMO-LUMO gap and greatly diminishes the DNA photocleavage efficiency. UVA light-induced photodegradation of selected MBAs reveals that methyl substitution at either 7 or 12 (or both) positions greatly enhances the degradation rate. Photodegradation of 7-MBA, 12-MBA, and DMBA yields products that are much less effective in mediating DNA cleavage. Photodegradation of other MBAs, exemplified by 5-MBA, yields a photooxidation product 5-MBA-7,12 quinone which is relatively stable under light and is a stronger DNA photocleaver than 5-MBA itself. The higher efficiency of DNA photocleavage for MBAs with methyl substitution at positions other than 7 or 12 is due, at least in part, to the formation of 7,12-quinone. Light-induced DNA single strand cleavage efficiency for several MBAs parallels the light-induced toxicity observed by other research groups, suggesting that light-induced DNA cleavage of MBAs are the source for phototoxicity. Since some PAHs such as coal tar are used commercially as creams, therapeutic agents, or ointments, or those roofers and asphalt workers that are subject to contamination with PAHs, the combination of PAHs and light (in the skin) may present a greater health risk to humans. PMID- 11896689 TI - Redox properties of Met(35) in neurotoxic beta-amyloid peptide. A molecular modeling study. AB - The beta-amyloid peptide (betaAP) is the principal component of plaque associated with the pathology of Alzheimer's disease. Part of its neurotoxicity appears to correlate with the ability of the peptide to reduce Cu(II) and form free radicals. Both processes are dependent on the presence and oxidizability of Met(35) in the C-terminus of the peptide but no mechanistic details on the reactions leading to Met oxidation are known. On the basis of previous studies with model peptides, we hypothesize that a one-electron oxidation of Met(35) in betaAP is facilitated through a neighboring group effect. Complexed to Cu(II) and/or in a lipid-mimicking environment, the solution structure of betaAP includes a large alpha-helical part. The solution NMR structure of betaAP1-40 in aqueous SDS micelles reveals an alpha-helix between residues 27 and 36, containing Met(35). In this helical C-terminus of betaAP, the peptide bond C=O group C-terminal of Ile(31) is located very close to the Met(35) sulfur and could stabilize a Met(35) sulfide radical cation through formation of an (S-O) three electron bond. In the present paper, we have computationally validated this hypothesis using Langevin dynamics methods to determine the collision frequency of the Met(35) thioether sulfur and the oxygen atoms of several peptide bonds in the betaAP sequence. Nanosecond time scale computations were carried out for four distinct betaAP congeners, betaAP26-40, betaAP26-36, betaAP26-40(Ile(31)Pro), betaAP40-26, and their respective Met(35)-sulfur-centered cation radicals. Here, betaAP26-40, betaAP26-40(Ile(31)Pro) and betaAP40-26 are representative fragments of the full length betaAP1-42 or betaAP42-1 sequence, respectively, whereas betaAP26-36 represents a unique betaAP sequence for which biological data are available. Initial structures of betaAP26-40, betaAP26-40(Ile(31)Pro), and betaAP26-36 were selected to be identical to that of the betaAP26-40 or betaAP26 36 sequence in full-length betaAP1-40. As the structures of betaAP40-26 and betaAP42-1 are not known, various initial conformations such as alpha-helix and antiparallel beta-sheet were selected for betaAP40-26. Our computational results show that betaAP26-40, representative for the same sequence in full-length betaAP1-42, has the highest tendency to form (S-O) bonds between Ile(31)C=O and Met(35)S. We conclude that native betaAP1-42 has a higher tendency to support Met(35) oxidation through (S-O) bond formation, consistent with the experimental observation that betaAP1-42 is more neurotoxic compared to the other investigated sequences. PMID- 11896690 TI - Identification of a new mutagenic polychlorinated biphenyl derivative in the Waka River, Wakayama, Japan, showing activation of an aryl hydrocarbon receptor dependent transcription. AB - Water samples from the Waka River, which runs through an area housing many chemical industry facilities in Wakayama, Japan, have been found to show significant mutagenicity, especially without a mammalian metabolic activation system (S9 mix) in the Salmonella typhimurium YG1024 strain. Mutagens in the river water were adsorbed to 3 kg of blue cotton, extracted with methanol/ammonia, and separated by several low- and high-pressure liquid chromatography steps with reversed-phase columns. One mutagen (0.6 mg), accounting for 50% of the total mutagenicity of the adsorbed materials, was isolated. On the basis of the mass, high-resolution mass, (1)H NMR and (13)C NMR spectra, the chemical was determined to have a polychlorinated biphenyl skeleton with nitro and amino substitution groups. Well-designed chemical synthesis of the putative mutagen revealed it to be 4-amino-3,3'-dichloro-5,4'-dinitrobiphenyl. This novel compound exerted strong mutagenicity without the S9 mix, inducing 66,000 and 140,000 revertants/nmol in S. typhimurium TA98 and YG1024, respectively. Moreover, this polychlorinated biphenyl derivative was proven to activate the human aryl hydrocarbon receptor-mediated transcription in a lac Z reporter gene assay with an efficiency almost the same as that of beta naphthoflavone, well-known to be a synthetic aryl hydrocarbon receptor agonist. It is possible that the mutagen is formed unintentionally via postemission modification of drainage water containing parent chemicals, such as 3,3' dichlorobenzidine or 3,3'-dichloro-4,4'-dinitrobiphenyl, which are known to be raw materials in the manufacture of polymers and dye intermediates in chemical plants. PMID- 11896691 TI - Optimization of the workup procedure for the analysis of 8-oxo-7,8-dihydro-2' deoxyguanosine with electrochemical detection. AB - The artifactual generation of the biomarker for oxidative stress, 8-oxo-7,8 dihydro-2'-deoxyguanosine (8-oxodG), during the workup procedure for its analysis is a difficult problem to solve, and the responsible factors are unclear. Here, peroxide removal and other antioxidant procedures during workup were compared using a limited amount of rat liver (50 mg) as starting material, with subsequent hydrolysis of 50 microg of DNA. A cold (0 degrees C) high salt GTC (4 M guanidine thiocyanate) nonphenol DNA extraction method was developed where DNA is quickly isolated. GSH (reduced glutathione) generated artifactual formation of 8-oxodG during the workup procedure, whereas H(2)O(2) removal using catalase, Fe(3+) removal and passivation using desferal, peroxide removal using glutathione peroxidase, ebselen and a peroxidase mimic lowered the 8-oxodG levels, all identifying peroxides as the responsible oxidants. Desferal was more protective when excluding Mg(2+) and Ca(2+) from buffers but was found to disturb the electrochemical detector when repeatedly injected five to six times, even at 100 microM. Addition of the OH(*) scavenger ethanol in all steps at 2% v/v had no protective effect. Zn(2+) was found necessary for efficient DNA hydrolysis using nuclease P(1), which was poor below 37 degrees C. Use of water substitutes was tested but inhibited DNA hydrolysis completely. H(2)(18)O could, however, work for mass spectrometry methods. Long-term (38 days) storage of 0.5% v/v Triton X 100 generated more 8-oxodG than Tween 20 when incubated with free dG. The cold GTC DNA extraction method was used for analysis of freshly isolated human lymphocytes/monocytes from 60 healthy men using catalase and TEMPO as antioxidants, giving a background level of 0.074 +/- 0.027 8-oxodG/10(5) dG (or 16 8-oxodG/10(8) nucleotides or 1943 8-oxodG/nuclei) which is probably the lowest value obtained yet. No increase with age was seen. Oxidation of dG to 8-oxodG during workup was found to fit a mathematically defined curve, and a calculated background level of 0.047 8-oxodG/10(5) dG was obtained. To obtain more reliable results it is recommended that control samples are included during the workup procedure, having an equal amount of cells (or DNA) as the exposed samples. PMID- 11896692 TI - Modified immunoenriched (32)P-HPLC assay for the detection of O(4)-ethylthymidine in human biomonitoring studies. AB - Increased excretion of ethylated DNA bases has been reported in the urine of cigarette smokers. To study DNA ethylation in the target organs of smokers, an immunoenriched (32)P-postlabeling assay for O(4)-ethylthymidine (O(4)-etT) was developed. O(4)-etT-3'-monophosphate (O(4)-etT-3'P) was synthesized, purified, and characterized by LC-MS, ESI-MS, and NMR. DNA was enzymatically digested to 2' deoxynucleoside-3'-monophosphate followed by immunoprecipitation of O(4)-etT-3'P using specific monoclonal antibodies. The immunoconjugate was washed by filtration, and O(4)-etT-3'P was recovered by ethanol treatment. The enriched O(4)-etT-3'P was labeled with [gamma-(32)P]ATP in the presence of T4 polynucleotide kinase at pH 6.8 to yield its 5'-labeled monophosphate and was subsequently resolved on RP-HPLC and detected with online detection of radioactivity. Adduct recovery was >80%, and the detection limit was approximately 500 amol. To further validate the method, O(4)-etT levels were determined in calf thymus DNA treated with N-ethyl-N-nitrosourea, and a dose dependent formation of O(4)-etT was observed. Furthermore, O(4)-etT was found to be present in the cells obtained from the lower respiratory tract by sputum induction of two out of four smokers but not in three nonsmokers. O(4)-etT is a poorly repaired promutagenic DNA lesion; thus, it could be of potential use for biomonitoring smoking-related DNA damage. Our improved assay was found to be sufficiently sensitive and specific to detect O(4)-etT in surrogate cells from cigarette smoke exposed humans. PMID- 11896693 TI - Conformational changes of a benzo[a]pyrene diol epoxide-N(2)-dG adduct induced by a 5'-flanking 5-methyl-substituted cytosine in a (Me)CG double-stranded oligonucleotide sequence context. AB - Mutations in p53 genes are one of the most common genetic alterations in human cancers. A disproportionate number of mutations are found in certain codons of the p53 gene, mostly at CpG dinucleotide sequences, which are highly methylated in human tissues. The reactivities of the mutagenic metabolite of benzo[a]pyrene, the bay region diol epoxide r7,t8-dihydroxy-t9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene (BPDE), to yield adducts with guanine at the exocyclic amino group (e.g., trans-anti-BPDE-N(2)-dG, or G*), are enhanced when the cytosine in CpG sequences in DNA is methylated at its 5-position ((Me)CpG). However, methylation may also affect the characteristics of these adducts, and we have therefore investigated whether adduct conformations are different in double stranded DNA in methylated (Me)CpG* and in unmethylated CpG* sequence contexts in the oligonucleotide model system duplex 5'-d(CCAT[(5X)C]GCTACC).d(GGTAGCGATGG) with X = H or -CH(3). The (-)-trans-adduct exhibits a striking conformational change from a minor groove structure external to the DNA duplex in the unmethylated CpG* sequence, to an intercalative conformation in the (Me)CG* sequence context. In contrast, the conformation of the stereoisomeric (+)-trans adduct is predominantly of the minor groove type in both the methylated and unmethylated sequences. These results indicate that methylation of CpG sequences may affect not only chemical reactivities of chemically reactive intermediates with DNA, but also the conformational properties of the DNA adducts formed. Thus, both factors must be considered in evaluating the effects of cytosine methylation in CpG sequences on the biological consequences of the DNA adducts formed. PMID- 11896694 TI - DNA tandem lesions containing 8-oxo-7,8-dihydroguanine and formamido residues arise from intramolecular addition of thymine peroxyl radical to guanine. AB - Exposure of aerated aqueous solutions of dinucleoside monophosphates bearing both a pyrimidine base and a guanine residue to ionizing radiation leads to the formation of 8-oxo-7,8-dihydroguanine/formamido tandem lesions (8-oxodG/dF). Recent evidence for the formation of the latter damage within isolated DNA emphasized the possible biological relevance of this class of lesions. Therefore, an extensive mechanistic study of the formation of 8-oxodG/dF was carried out with thymine and guanine containing dinucleoside monophosphates (dGpdT and dTpdG). First, the peroxyl radical-induced degradation of guanine within dGpdT and dTpdG was studied in order to assess the possibility of intramolecular electron transfer between guanine and thymine peroxyl radicals. Then, the formation of a series of tandem lesions involving a formamido residue, thymine glycols, 8-oxodG, and oxazolone was monitored within aerated aqueous solutions of dTpdG and dGpdT exposed to gamma-radiation. The absence of formation of tandem lesions other than 8-oxodG/dF in significant yield led to us propose a new mechanism involving addition of the thymine peroxyl radical to the guanine moiety. This received support from (18)O labeling experiments. PMID- 11896695 TI - Molecular metal sulfide cluster model for substrate binding to oil-refinery hydrodesulfurization catalysts. AB - Reaction between [(eta5-Cp')3Mo3S4]+ and [Ni(1,5-cod)2] (Cp' = methylcyclopentadienyl; 1,5-cod = 1,5-cyclooctadiene) in THF at ambient temperature yielded a coordinatively unsaturated cubane-like cluster cation, [(eta5-Cp')3Mo3S4Ni]+. The ligand sphere at the Ni atom could be saturated by coordinating dimethyl sulfide, diethyl sulfide, di(tert-butyl) sulfide, tetrahydrothiophene, thiochroman-4-ol, 1,4-dithiane, pyridine, quinoline, or 4,4' bipyridine. The products structurally model a mode of substrate coordination on proposed binding sites of heterogeneous MoNi sulfide hydrotreating catalysts. No stable coordination compounds could be isolated for thiophene derivatives. X-ray crystal structures are reported for the ligand-bridged dicluster compounds [[(eta5-Cp')3Mo3S4Ni]2(mu-C4H4S2)][pts]2 (C4H8S2 = 1,4-dithiane) and [[(eta5 Cp')3Mo3S4Ni]2(mu-bipy)][pts]2 (bipy = 4,4'-bipyridine). PMID- 11896696 TI - A simultaneous reduction, substitution, and self-assembly reaction under hydrothermal conditions afforded the first diiodopyridine copper(I) coordination polymer. AB - A simultaneous reduction of copper(II) to copper(I) by pyridinecarboxylate and the substitution of carboxylato groups by iodo nucleophiles in a self-assembly process under hydrothermal conditions afforded a new iodine-inclusion coordination polymer [CuI(C5H3NI2)*1/2I2] 1. The synthetic studies of the substitution process produced a new supramolecular compound [IC5H3NCOOH] 2 and revealed that the catalytic properties of copper ions in redox and substitution reactions under hydrothermal conditions are attractive. Crystal data for [CuI(C5H3NI2)*1/2I2]: triclinic, space group P1; cell dimensions a = 4.216(1) A, b = 11.254(2) A, c = 12.196(2) A, alpha = 80.34(3) degrees, beta = 88.44(3) degrees, gamma = 83.10(3); V = 566.2(2) A(3), Z = 2. Crystal data for [IC(5)H(3)NCOOH]: monoclinic, space group P2(1)/c; cell dimensions a = 5.041(1) A, b = 17.313(2) A, c = 8.639(1) A, beta = 95.042(2) degrees; V = 751.02(13) A(3), Z = 4. PMID- 11896697 TI - Oxidative degradation of beta-diketiminate ligand in copper(II) and zinc(II) complexes. AB - Copper(II) and zinc(II) complexes supported by a popular beta-diketiminate ligand (1(-), 2-mesitylamino-4-mesitylimino-2-pentene), [CuII(1)(AcO)] and [[ZnII(1)]2(mu-MeO)(mu-AcO)], have been demonstrated to undergo an oxidative degradation to give a ketone diimine derivative (2) under aerobic conditions. The crystal structures of the mononuclear copper(II) and dinuclear zinc(II) complexes of the beta-diketiminate ligand as well as the copper(II) complex of the modified ligand have been determined by X-ray crystallographic analysis. Mechanism for the oxidative degradation reaction of the beta-diketiminate ligand is also discussed. PMID- 11896698 TI - Evidence of desulfurization in the oxidative cyclization of thiosemicarbazones. Conversion to 1,3,4-oxadiazole derivatives. AB - The addition of pyridine-2-carbaldehyde 4N-methylthiosemicarbazone (C8H10N4S) to an aqueous solution of copper(II) nitrate yields [[Cu(C8H9N4S)(NO3)]2] (1). This complex consists of centrosymmetric dinuclear entities containing square pyramidal copper(II) ions bridged through the sulfur thioamide atoms. The oxidation of 1 with KBrO3 or KIO3 gives rise to a compound with formula [[Cu(C8H8N4O)(H2O)2(SO4)]2]*2H2O (2) (C8H8N4O = 2-methylamino-5-pyridin-2-yl 1,3,4-oxadiazole). The structure of 2 is made up of centrosymmetric dimers where the copper(II) ions exhibit a distorted octahedral coordination and are connected by the oxadiazole moiety. The metal ions in 2 can be removed by addition of K4[Fe(CN)6], and then the oxadiazole ligand can be isolated and recrystallized as (C8H8N4O)*3H2O (3). PMID- 11896699 TI - Pressure-controlled plasticity of Cu(H2O)6(2+) complex and phase transition in Cs2Cu(ZrF6)2*6H2O. AB - The X-band EPR study of a polycrystalline Cs2Cu(ZrF6)2*6H2O demonstrates a feature of plasticity of the Jahn-Teller Cu(H2O)6 complex in the crystal lattice of this compound. The temperature- and pressure-induced evolution of the spectra shows that the copper complex is extremely sensitive to these factors, which due to the ferroelastic properties of the compound studied modify the internal tetragonal and orthorhombic strains acting on the complex. It is supported by the analysis of the temperature dependencies of the principal values of the g-factor under various pressures, indicating that the complex varies its shape adapting it to the varied conditions. A pressure-induced phase transition is discovered. PMID- 11896700 TI - Modulating the reduction potential of mononuclear cobalt(II) complexes via selective deprotonation of tris[(2-benzimidazolyl)methyl]amine. AB - Remote site deprotonation of the coordinated tripodal ligand, tris((2 benzimidazolyl)methyl)amine, was examined using electronic spectroscopy and electrochemistry techniques. The solid-state structures [CoH(3)1(tba)(NCS)]+ and [CoH(2)1(tba)(NCS)] are reported. These complexes crystallized in the triclinic space group P1 [a = 13.3043(2) A, b = 13.8019(2) A, c = 14.1322(2) A, alpha = 63.6670(10) degrees, beta = 68.0590(10) degrees, gamma = 81.8960 degrees; Z = 2] and the monoclinic space group P2(1)/n [a = 15.3530(9) A, b = 11.0645(6) A, c = 19.1319(10) A, beta = 105.6750(10) degrees; Z = 4], respectively. Preliminary results suggest that selective and reversible deprotonation of coordinated benzimidazolyl ligands can tune the reduction potential of several isostructural cobalt(II) complexes. PMID- 11896701 TI - The first designed syntheses of bis-dimetal molecules in which the bridges are diamidate ligands. AB - The first deliberate syntheses of molecules in which pairs of quadruply bonded Mo2 units are bridged by N,N'-diarylterephthaloyldiamidate (aryl = Ph, m-CF3Ph) ligands are described. The addition of neutral N,N'-diarylterephthaloyldiamide to 2 equiv of [Mo2(DAniF)3(MeCN)2]+ (DAniF = N,N'-di-p-anisylformamidinate) followed by the introduction of excess H3CO- in MeCN results in the formation of (DAniF)3Mo2[(C6H5)NC(O)C6H4(O)CN(C6H5)]Mo2(DAniF)3 (1) and (DAniF)3Mo2[[(m CF3)C6H5]NC(O)C6H4(O)CN[(m-CF3)C6H5]]Mo2(DAniF)3 (2). The DeltaE1/2 for the oxidation of each Mo2 unit is greater for these terephthaloyldiamidate-bridged molecules (approximately 100 mV) than for the analogous terephthalate-bridged compound (approximately 60 mV). Variation in the nature of the substituents on the diamidate nitrogen atoms offers a means to fine-tune the oxidation potentials of the Mo2 units. PMID- 11896702 TI - Reduction of [VO2(ma)2]- and [VO2(ema)2]- by ascorbic acid and glutathione: kinetic studies of pro-drugs for the enhancement of insulin action. AB - To shed light on the role of V(V) complexes as pro-drugs for their V(IV) analogues, the kinetics of the reduction reactions of [VO2(ma)2]- or [VO2(ema)2]- (Hma = maltol, Hema = ethylmaltol), with ascorbic acid or glutathione, have been studied in aqueous solution by spectrophotometric and magnetic resonance methods. EPR and 51V NMR studies suggested that the vanadium(V) in each complex was reduced to vanadium(IV) during the reactions. All the reactions studied showed first-order kinetics when the concentration of ascorbic acid or glutathione was in large excess and the observed first-order rate constants have a linear relationship with the concentrations of reductant (ascorbic acid or glutathione). Potentiometric results revealed that the most important species in the neutral pH range is [VO2(L)2]- for the V(V) system where L is either ma- or ema-. An acid dependence mechanism was proposed from kinetic studies with varying pH and varying maltol concentration. The good fits of the second order rate constant versus pH or the total concentration of maltol, and the good agreement of the constants obtained between fittings, strongly supported the mechanism. Under the same conditions, the reaction rate of [VO2(ma)2]- with glutathione is about 2000 times slower than that of [VO2(ma)2]- with ascorbic acid, but an acid dependence mechanism can also be used to explain the results for the reduction with glutathione. Replacing the methyl group in maltol with an ethyl group has little influence on the reduction rate with ascorbic acid, and the kinetics are the same no matter whether [VO2(ma)2]- or [VO2(ema)2]- is reduced. PMID- 11896704 TI - Reactivity of the heterometallic clusters [HMCo3(CO)12] and [Et4N][MCo3(CO)12] (M = Fe, Ru) toward phosphine selenides, including selenium transfer. Crystal structures of [HRuCo3(CO)7(mu-CO)3(mu-dppy)], [MCo2(mu3-Se)(CO)7(mu-dppy)], and [RuCo2(mu3-Se)(CO)7(mu-dppm)] [dppy = Ph2(2-C5H4N)P, dppm = Ph2PCH2PPh2]. AB - The reactivity of [HMCo3(CO)12] and [Et4N][MCo3(CO)12] (M = Fe, Ru) toward phosphine selenides such as Ph3PSe, Ph2P(Se)CH2PPh2, Ph2(2-C5H4N)PSe, Ph2(2 C4H3S)PSe, and Ph2[(2-C5H4N)(2-C4H2S)]PSe has been studied with the aim to obtain new selenido-carbonyl bimetallic clusters. The reactions of the hydrido clusters give two main classes of products: (i) triangular clusters with a mu3-Se capping ligand of the type [MCo2(mu3-Se)(CO)(9-x)L(y)] resulting from the selenium transfer (x = y = 1, 2, with L = monodentate ligand; x = 2, 4, and y = 1, 2, with L = bidentate ligand) (M = Fe, Ru) and (ii) tetranuclear clusters of the type [HMCo3(CO)12xL(y)] obtained by simple substitution of axial, Co-bound carbonyl groups by the deselenized phosphine ligand. The crystal structures of [HRuCo3(CO)7(mu-CO)3(mu-dppy)] (1), [MCo2(mu3-Se)(CO)7(mu-dppy)] (M = Fe (16) or Ru (2)), and [RuCo2(mu3-Se)(CO)7(mu-dppm)] (12) are reported [dppy = Ph2(2 C5H4N)P, dppm = Ph2PCH2PPh2]. Clusters 2, 12, and 16 are the first examples of trinuclear bimetallic selenido clusters substituted by phosphines. Their core consists of metal triangles capped by a mu3-selenium atom with the bidentate ligand bridging two metals in equatorial positions. The core of cluster 1 consists of a RuCo3 tetrahedron, each Co-Co bond being bridged by a carbonyl group and one further bridged by a dppy ligand. The coordination of dppy in a pseudoaxial position causes the migration of the hydride ligand to the Ru(mu-H)Co edge. In contrast to the reactions of the hydrido clusters, those with the anionic clusters [MCo3(CO)12]- do not lead to Se transfer from phosphorus to the cluster but only to CO substitution by the deselenized phosphine. PMID- 11896703 TI - Ruthenium complexes of analogues of the antitumor antibiotic streptonigrin. AB - The complexes Ru(L1-CH3)(CO)3Cl, RuL2(CO)2Cl2, and RuL3(CO)2Cl2 (L1= 6-methoxy 5,8-quinolinedione, L2 = 7-amino-6-methoxy-5,8-quinolinedione, L3 = 6,6' dimethoxycarbonyl-2,2'-bipyridine) were prepared by reaction of L1-L3 with the tricarbonyldichlororuthenium(II) dimer. L1-L3 act as bidentates through the ortho oxygen atoms, the pyridyl nitrogen and the adjacent quinone oxygen, and the bipyridyl nitrogens, respectively. RuL3(CO)2Cl2 is characterized by X-ray crystallography. 15N NMR correlation spectra give upfield shifts of around 60 ppm for the pyridyl nitrogens that are coordinated to the metal, while 13C NMR correlation spectra give a downfield shift of 10 ppm for the quinone carbonyl group that is coordinated to the metal. The electrochemistry of RuL2(CO)2Cl2 is examined, and the implications for the formation of metal complexes of the antitumor antibiotic streptonigrin, which cleaves DNA in the presence of metal ions, are discussed. PMID- 11896705 TI - Nickel complexes of bidentate [S2] and [SN] borato ligands. AB - Nickel(II) complexes of the monoanionic borato ligands [Ph2B(CH2SCH3)2] (abbreviated Ph2Bt), [Ph2B(CH2S(t)Bu)2] (Ph2Bt(tBu)), [Ph2B(1 pyrazolyl)(CH2SCH3)], and [Ph2B(1-pyrazolyl)(CH2S(t)Bu)] have been prepared and characterized. While [Ph2Bt] formed the square planar homoleptic complex, [Ph2Bt]2Ni, the larger [S2] ligand with tert-butyl substituents, [Ph2BttBu], yielded an unexpected organometallic derivative, [Ph2Bt(tBu)]Ni(eta2-CH2SBut), resulting from B-C bond rupture. The analogous thiametallacycle derived from the [S3] ligand, [PhB(CH2S(t)Bu)3] (PhTt(tBu)), has been structurally authenticated (Schebler, P. J.; Mandimutsira, B. S.; Riordan, C. G.; Liable-Sands, L.; Incarvito, C. D.; Rheingold, A. L. J. Am. Chem. Soc. 2001, 123, 331). The [SN] borato ligands formed exclusively the cis stereoisomers upon reaction with Ni(II) sources, [Ph2B(1-pyrazolyl)(CH2SR)]2Ni. Analysis of the Ni(II/I) reduction potentials by cyclic voltammetry revealed a approximately 600 mV anodic shift upon replacement of two thioether donors ([Ph2Bt]2Ni) with two pyrazolyl donors ([Ph2B(1-pyrazolyl)(CH2SCH3)]2Ni) consistent with the all thioether environment stabilizing the lower oxidation state of nickel. PMID- 11896706 TI - Synthesis, structure, and fluorescence of the novel cadmium(II)-trimesate coordination polymers with different coordination architectures. AB - Three novel complexes, Cd3tma2*13H2O (1), Cd3tma2*dabco*2H2O (2), and Cd3Htma3*8H2O (3) (tma = trimesate), of cadmium(II)-trimesate coordination polymers are obtained from hydrothermal reaction. 1 (C18H32O25Cd3) crystallizes in the monoclinic C2/c space group [a = 18.985(2) A, b = 7.3872(6) A, c = 20.432(2) A, = 97.1660(10), and Z = 4]. 2 (C24H22N2O14Cd3) crystallizes in the monoclinic P2(1)/c space group [a = 10.1323(2) A, b = 19.5669(5) A, c = 13.15880(10) A, = 108.9810(10), and Z = 4]. 3 (C27H28O26Cd3) belongs to the trigonal P31c space group [a = 15.7547(3) A, b = 15.7547(3) A, c = 7.93160(10) A, and Z = 2]. The Cd(II) centers in the three complexes are bridged by tma ligands in the coordination fashion of unidentate, bridging unidentate, bidentate, chelating bis-bidentate, chelating/bridging bis-bidentate, or chelating/bridging bidentate to form the T-shaped molecular bilayer motif for 1, chicken-wire-like motif for 2, and honeycomb-like porous structure for 3, respectively, in which the T-shaped molecular bilayer motif and chicken-wire-like motif are further interlinked in interdigitating or alternating fashion to construct the different coordination architectures. These three complexes exhibit strong fluorescent emission bands at 355 nm (lambda(ex) = 220 nm) for 1, 437 nm (lambda(ex) = 365 nm) for 2, and 353 nm (lambda(ex) = 218 nm) for 3 in the solid state at room temperature. PMID- 11896707 TI - (Eta5-pentamethylcyclopentadienyl)rhodium and -iridium complexes with weakly and strongly coordinating anions: isolation and first X-ray molecular structures of the tris(solvent) complexes [(C5Me5)M(acetone)2(H2O)][BF4]2 (M = Rh, Ir). AB - Several novel pentamethylcyclopentadienyl complexes of general formula [(C5Me5)IrL3][BF4]2 were prepared including the tris(solvent) precursors [(C5Me5)M(acetone)2(H2O)][BF4]2 (M = Rh, Ir) (1a,b). The X-ray molecular structures of 1a,b were determined at low temperature. Complexes 1a,b are isostructural, and both compounds crystallize in the monoclinic space group P2(1)/c with a = 10.157(3) A, b = 14.038(9) A, c = 16.335(2) A, beta = 99.73(2) degrees, and Z = 4 for 1a and with a = 10.107(9) A, b = 13.994(16) A, c = 15.996(34) A, beta = 99.61(12) degrees, and Z = 4 for 1b. The coordinated water molecule is hydrogen bonded to both BF4(-) anions. Reaction of 1a,b with pyridine (py) afforded the related tris(pyridine) complexes [(C5Me5)M(eta1-(N)-py)3][BF4]2 (M = Rh, Ir) (2a,b). Complex 2b was characterized by X-ray crystallography, monoclinic space group P2(1)/c with a = 8.665(3) A, b = 19.687(7) A, c = 18.408(5) A, beta = 94.17(3) degrees, and Z = 4. Moreover, we prepared the novel neutral compounds (C5Me5)M(eta2-NO3)(eta1-NO3) (M = Rh, Ir) (4a,b) where the anions are bonded to the metal center instead of a coordinating solvent as confirmed by X-ray study on the iridium complex 4b. The latter crystallizes in the orthorhombic space group Pcab with a = 13.032(4) A, b = 14.370(11) A, c = 14.839(18) A, and Z = 8. PMID- 11896708 TI - Two new terpyridine dimanganese complexes: a manganese(III,III) complex with a single unsupported oxo bridge and a manganese(III,IV) complex with a dioxo bridge. Synthesis, structure, and redox properties. AB - Two new terpyridine dimanganese oxo complexes [Mn(2)(III,IV)(mu O)(2)(terpy)(2)(CF(3)CO(2))(2)](+) (3) and [Mn(2)(III,III)(mu O)(terpy)(2)(CF(3)CO(2))(4)] (4) (terpy = 2,2':6,2' '-terpyridine) have been synthesized and their X-ray structures determined. In contrast to the corresponding mixed-valent aqua complex [Mn(2)(III,IV)(mu O)(2)(terpy)(2)(H(2)O)(2)](3+) (1), the two Mn atoms in 3 are not crystallographically equivalent. The neutral binuclear monooxo manganese(III,III) complex 4 exhibits two crystallographic forms having cis and trans configurations. In the cis complex, the two CF(3)CO(2)(-) ligands on each manganese adopt a cis geometry to each other; one CF(3)CO(2)(-) is trans to the oxygen of the oxo bridge while the second is cis. In the trans complex, the two coordinated CF(3)CO(2)(-) have a trans geometry to each other and are cis to the oxo bridge. The electrochemical behavior of 3 in organic medium (CH(3)CN) shows that this complex could be oxidized into its corresponding stable manganese(IV,IV) species while its reduced form manganese(III,III) is very unstable and leads by a disproportionation process to Mn(II) and Mn(IV) complexes. Complex 4 is only stable in the solid state, and it disproportionates spontaneously in CH(3)CN solution into the mixed-valent complex 3 and the mononuclear complex [Mn(II)(terpy)(2)](2+) (2), thereby preventing the observation of its electrochemical behavior. PMID- 11896709 TI - Pyrazolyl-bridged iridium dimers. 18.(1) influence of metal-metal bonding on the geometry of diiridium(II) adducts and hydrido-diiridium complexes formed from the diiridium(I) prototype [Ir(mu-pz)(PPh(3))(CO)](2) (pzH = Pyrazole) by dihydrogen addition or protonation. AB - Slow uptake of molecular dihydrogen by the diiridium(I) prototype [Ir(mu pz)(PPh(3))(CO)](2) (1: pzH = pyrazole) is accompanied by formation of a 1,2 dihydrido-diiridium(II) adduct [IrH(mu-pz)(PPh(3))(CO)](2) (2), for which an X ray crystal structure determination reveals that (unlike in 1) the PPh(3) ligands are axial, with the hydrides occupying trans coequatorial positions across the Ir Ir bond (2.672 A). Reaction with CCl(4) effects hydride replacement in 2, affording the monohydride Ir(2)H(Cl)(mu-pz)(2)(PPh(3))(2)(CO)(2) (3) in which Ir Ir = 2.683 A. At one metal center, H is equatorial and PPh(3) is axial, while at the other, Cl is axial as is found in the symmetrically substituted product [Ir(mu-pz)(PPh(3))(CO)Cl](2) (4) (Ir-Ir = 2.754 A) that is formed by action of CCl(4) on 1. Treatment of 1 with I(2) yields the diiodo analogue 5 of 4, which reacts with LiAlH(4) to afford the isomorph Ir(2)H(I)(mu-pz)(2)(PPh(3))(2)(CO)(2) (6) of 3 (Ir-Ir = 2.684 A). Protonation (using HBF(4)) of 1 results in formation of the binuclear cation Ir(2)H(mu-pz)(2)(PPh(3))(2)(CO)(2)(+) (7: BF(4)(-) salt), which shows definitive evidence (from NMR) for a terminally bound hydride in solution (CH(2)Cl(2) or THF), but 7 crystallizes as an axially symmetric unit in which Ir-Ir = 2.834 A. Reaction of 7 with water or wet methanol leads to isolation of the cationic diiridium(III) products [Ir(2)H(2)(mu-OX)(mu pz)(2)(PPh(3))(2)(CO)(2)]BF(4) (8, X = H; 9, X = Me). PMID- 11896711 TI - Preparation and structural characterization of (Me(3)SiNSN)(2)Se, a new synthon for sulfur-selenium nitrides. AB - The reaction of (Me(3)SiN)(2)S with SeCl(2) (2:1 ratio) in CH(2)Cl(2) at -70 degrees C provides a route to the novel mixed selenium-sulfur-nitrogen compound (Me(3)SiNSN)(2)Se (1). Crystals of 1 are monoclinic and belong the space group P2(1)/c, with a = 7.236(1) A, b = 19.260(4) A, c = 11.436(2) A, beta = 92.05(3) degrees, V = 1592.7(5) A(3), Z = 4, and T = -155(2) degrees C. The NSNSeNSN chain in 1 consists of Se-N single bonds (1.844(3) A) and S=N double bonds (1.521(3) 1.548(3) A) with syn and anti geometry at the N=S=N units. The N-Se-N bond angle is 91.8(1) degrees. The EI mass spectrum shows a molecular ion with good agreement between the observed and calculated isotopic distributions. The (14)N NMR spectrum exhibits two resonances at -65 and -77 ppm. Both (13)C and (77)Se NMR spectra show single resonances at 0.83 and 1433 ppm, respectively. The reaction of 1 with an equimolar amount of SeCl(2) produces 1,5-Se(2)S(2)N(4) (2) in a good yield, and that of (Me(3)SiNSN)(2)S with SCl(2) affords S(4)N(4) (3), but the reactions of (Me(3)SiNSN)(2)Se with SCl(2) and (Me(3)SiNSN)(2)S with SeCl(2) result in the formation of a mixture of 2 and 3. A likely reaction pathway involves the intermediate formation of E(2)N(2) fragments (E = S, Se). PMID- 11896710 TI - DFT investigation of structural, electronic, and catalytic properties of diiron complexes related to the [2Fe](H) subcluster of Fe-only hydrogenases. AB - Hydrogenases catalyze the reversible oxidation of dihydrogen to protons and electrons. The structures of two Fe-only hydrogenases have been recently reported [Peters, J. W.; Lanzilotta, W. N.; Lemon, B. J.; Seefeldt, L. C. Science 1998, 282, 1853-1858. Nicolet, Y.; Piras, C.; Legrand, P.; Hatchikian, E. C.; Fontecilla-Camps, J. C. Structure 1999, 7, 13-23], showing that the likely site of dihydrogen activation is the so-called [2Fe](H) cluster, where each Fe ion is coordinated by CO and CN(-) ligands and the two metals are bridged by a chelating S-X(3)-S ligand. Moreover, the presence of a water molecule coordinated to the distal Fe2 center suggested that the Fe2 atom could be a suitable site for binding and activation of H(2). In this contribution, we report a density functional theory investigation of the structural and electronic properties of complexes derived from the [(CO)(CH(3)S)(CN)Fe(II)(mu-PDT)Fe(II)(CO)(2)(CN)](-1) species, which is related to the [2Fe](H) cluster observed in Fe-only hydrogenases. Our results show that the structure of the [2Fe](H) cluster observed in the enzyme does not correspond to a stable form of the isolated cluster, in the absence of the protein. As a consequence, the reactivity of [(CO)(CH(3)S)(CN)Fe(II)(mu-PDT)Fe(II)(CO)(2)(CN)](-1) derivatives in solution may be expected to be quite different from that of the active site of Fe-only hydrogenases. In fact, the most favorable path for H(2) activation involves the two metal atoms and one of the bridging S atoms and is associated with a very low activation energy (5.3 kcal mol(-1)). The relevance of these observations for the catalytic properties of Fe-only hydrogenases is discussed in light of available experimental and theoretical data. PMID- 11896712 TI - Analysis of paramagnetic NMR spectra of triple-helical lanthanide complexes with 2,6-dipicolinic acid revisited: a new assignment of structural changes and crystal-field effects 25 years later. AB - Variable-temperature (1)H and (13)C NMR measurements of the D(3)-symmetrical triple-helical complexes [Ln(L1-2H)(3)](3)(-) (L1 = pyridine-2,6-dicarboxylic acid; Ln = La-Lu) show evidence of dynamic intermolecular ligand-exchange processes whose activation energies depend on the size of the metal ion. At 298 K, the use of diastereotopic probes in [Ln(L3-2H)(3)](3)(-) (L3 = 4-ethyl pyridine-2,6-dicarboxylic acid) shows that fast intramolecular P <==> M interconversion between the helical enantiomers occurs on the NMR time scale. Detailed analyses of the paramagnetic NMR hyperfine shifts according to crystal field independent techniques demonstrate the existence of two different helical structures, one for large lanthanides (Ln = La-Eu) and one for small lanthanides (Ln = Tb-Lu), in complete contrast with the isostructurality proposed 25 years ago. A careful reconsideration of the original crystal-field-dependent analysis shows that an abrupt variation of the axial crystal-field parameter A(0)2 parallels the structural change leading to some accidental compensation effects that prevent the detection of structural variations according to the classical one-nucleus method. Crystal structures in the solid state and density functional theory calculations in the gas phase provide structural models that rationalize the paramagnetic NMR data. A regular triple-helical structure is found for small lanthanides (Ln = Tb-Lu) in which the terdentate chelating ligands are rigidly tricoordinated to the metals. A flexible and distorted structure is evidenced for Ln = La-Eu in which the central pyridine rings interact poorly with the metal ion. The origin of the simultaneous variation of structural parameters and crystal-field and hyperfine constants near the middle of the lanthanide series is discussed together with the use of crystal-field-independent techniques for the interpretation of paramagnetic NMR spectra in axial lanthanide complexes. PMID- 11896713 TI - Synthesis and structural characterization of 2,6-dimesitylphenyl complexes of scandium, ytterbium, and yttrium. AB - The molecular structures of a number of 2,6-dimesitylphenyl-based (2,6 dimesitylphenyl = Dmp) complexes of the group 3 elements scandium and yttrium as well as of the lanthanide element ytterbium are reported. Reaction of 1 equiv of DmpLi with 1 equiv of MCl(3) (M = Sc, Yb, Y) in tetrahydrofuran at room temperature followed by crystallization from toluene/hexanes at -30 degrees C produces DmpMCl(2)(THF)(2) (M = Sc: 1; M = Yb: 2) and DmpMCl(2)(THF)(3) (M = Y: 3), respectively. The one-pot reaction of DmpLi with 1 equiv of YbCl(3) in tetrahydrofuran at room temperature followed by addition of 1 equiv of KO(t)Bu produces the heterobimetallic monoalkoxide complex DmpYb(THF)(O(t)Bu)(mu Cl)(2)Li(THF)(2) (4), which was crystallized from toluene/tetrahydrofuran (20:1) at -30 degrees C. Crystal data for 1: monoclinic, P2(1)/n; T = 203 K; a = 10.178(3) A; b = 15.468(3) A; c = 20.132(5) A; beta = 101.85(3) degrees; V = 3102.0(17) A(3); Z' = 4; D(calcd) = 1.228 g cm(-3); R(1) = 5.89%. Crystal data for 2: monoclinic, P2(1)/n; T = 173 K; a = 10.2447(7) A; b = 15.5683(12) A; c = 20.0979(14) A; beta = 101.749(4) degrees; V = 3238.3(5) A(3); Z' = 4; D(calcd) = 1.485 g cm(-3); R(1) = 4.32%. Crystal data for 3: monoclinic, P2(1)/n; T = 203 K; a = 15.950(3) A; b = 11.865(2) A; c = 18.254(3) A; beta = 92.323(3) degrees; V = 3451.9(10) A(3); Z' = 4; D(calcd) = 1.327 g cm(-)(3); R(1) = 4.43%. Crystal data for 4: triclinic, P1; T = 193 K; a = 10.2252(2) A; b = 11.3497(2) A; c = 18.5814(2) A; alpha = 98.7353(6) degrees; beta = 102.8964(6) degrees; gamma = 94.8058(5) degrees; V = 2062.09(5) A(3); Z' = 2; D(calcd) = 1.375 g cm(-3); R(1) = 4.56%. The molecular structures of 1-3 feature monomeric complexes with distorted trigonal-bipyramidal (1 and 2) or octahedral (3) coordination geometry about the metal atom, with the two chlorine atoms occupying the axial positions. 4 represents the first example of an alkoxide derivative of a terphenyl lanthanide complex. The molecular structure of the ate complex 4 exhibits a heavily distorted trigonal-bipyramidal coordination polyhedron about the ytterbium atom, with one of the mu-chlorine atoms and the oxygen atom of the tetrahydrofuran ligand representing the axial positions of the trigonal bipyramidal arrangement. A terminal alkoxide ligand is another main feature of the molecular structure of complex 4. PMID- 11896714 TI - Quantitative reactivity model for the hydration of carbon dioxide by biomimetic zinc complexes. AB - A quantitative structure-reactivity relationship has been derived from the results of B3LYP/6-311+G calculations on the hydration of carbon dioxide by a series of zinc complexes designed to mimic carbonic anhydrase. The reaction mechanism found is general for all complexes investigated. The reaction exhibits a low (4-6 kcal/mol) activation energy and is exothermic by about 8 kcal/mol. The calculations suggest an equilibrium between Lipscomb and Lindskog intermediates. The effectiveness of the catalysis is a function of the nucleophilicity of the zinc-bound hydroxide and the nucleofugicity of the zinc-bound bicarbonate. Hydrogen bridging of the bicarbonate to NH moieties in the ligands also plays an important role. PMID- 11896715 TI - Thermodynamics, kinetics, and mechanism of the stepwise dissociation and formation of Tris(L-lysinehydroxamato)iron(III) in aqueous acid. AB - pK(a) values for the hydroxamic acid, alpha-NH(3)(+), and epsilon-NH(3)(+) groups of L-lysinehydroxamic acid (LyHA, H(3)L(2+)) were found to be 6.87, 8.89, and 10.76, respectively, in aqueous solution (I = 0.1 M, NaClO(4)) at 25 degrees C. O,O coordination to Fe(III) by LyHA is supported by H(+) stoichiometry, UV-vis spectral shifts, and a shift in nu(CO) from 1648 to 1592 cm(-1) upon formation of mono(L-lysinehydroxamato)tetra(aquo)iron(III) (Fe(H(2)L)(H(2)O)(4)(4+)). The stepwise formation of tris(L-lysinehydroxamato)iron(III) from Fe(H(2)O)(6)(3+) and H(3)L(2+) was characterized by spectrophotometric titration, and the values for log beta(1), log beta(2), and log beta(3) are 6.80(9), 12.4(2), and 16.1(2), respectively, at 25 degrees C and I = 2.0 M (NaClO(4)). Stopped-flow spectrophotometry was used to study the proton-driven stepwise ligand dissociation kinetics of tris(L-lysinehydroxamato)iron(III) at 25 degrees C and I = 2.0 M (HClO(4)/NaClO(4)). Defining k(n) and k(-n) as the stepwise ligand dissociation and association rate constants and n as the number of bound LyHA ligands, k(3), k(-3), k(2), k(-2), k(1), and k(-1) are 3.0 x 10(4), 2.4 x 10(1), 3.9 x 10(2), 1.9 x 10(1), 1.4 x 10(-1), and 1.2 x 10(-1) M(-1) s(-1), respectively. These rate and equilibrium constants are compared with corresponding constants for Fe(III) complexes of acetohydroxamic acid (AHA) and N methylacetohydroxamic acid (NMAHA) in the form of a linear free energy relationship. The role of electrostatics in these complexation reactions to form the highly charged Fe(LyHA)(3)(6+) species is discussed, and an interchange mechanism mediated by charge repulsion is presented. The reduction potential for tris(L-lysinehydroxamato)iron(III) is -214 mV (vs. NHE), and a comparison to other hydroxamic acid complexes of Fe(III) is made through a correlation between E(1/2) and pFe. PMID- 11896716 TI - Tribromohydroxyphosphonium hexafluorometalate: synthesis, spectroscopic characterization, and crystal structure of Br(3)POH(+)AsF(6)(-). AB - The reaction of tribromophosphine oxide in the superacidic systems XF/MF(5) (X = H, D; M = As, Sb) leads to tribromohydroxyphosphonium hexafluorometalates. The structure was successfully elucidated in the case of tribromohydroxyphosphonium hexafluoroarsenate. Br(3)POH(+)AsF(6)(-) crystallizes in the orthorhombic space group Pnma (No. 53 with a = 1292.5(1) pm, b = 871.6(1) pm, and c = 845.0(1) pm) with four formula units per cell. The Raman, IR, (1)H NMR, and (31)P NMR spectra of Br(3)POX(+)MF(6)(-) (X = H, D; M = As, Sb) are reported. PMID- 11896717 TI - Synthesis, structure, and magnetic properties of tetranuclear cubane-like and chain-like iron(II) complexes based on the N(4)O pentadentate dinucleating ligand 1,5-bis[(2-pyridylmethyl)amino]pentan-3-ol. AB - The tetranuclear complexes [Fe(4)(pypentO)(pym)(3)(Oac)(NCS)(3)] x 1.5EtOH (1), [Fe(4)(pypentO)(pym)(Oac)(2)(NCS)(2)(MeO)(2)(H(2)O)] x H(2)O (2), [Fe(2)(pypentO)(NCO)(3)](2) (3), and [Fe(2)(pypentO)(N(3))(3)](2) (4) have been prepared, and their structure and magnetic properties have been studied (pypentOH = 1,5-bis[(2-pyridylmethyl)amino]pentan-3-ol, pymH = 2-pyridylmethanol). The X ray diffraction analysis of 1 (C(43)H(53)N(10)O(7.5)S(3)Fe(4), monoclinic, P2(1)/n, a = 11.6153(17) A, b = 34.391(17) A, c = 14.2150(18) A, beta = 110.88(5) degrees, V = 5305(3) A(3), Z = 4) and 2 (C(31)H(45)N(7)O(10)S(2)Fe(4), monoclinic, C2/c, a = 19.9165(17) A, b = 21.1001(12) A, c = 21.2617(19) A, beta = 104.441(10) degrees, V = 8652.7(12) A(3), Z = 8) showed a Fe(4)O(4) cubane-like arrangement of four iron(II) atoms, four mu(3)-O bridging ligands, one (1) or two (2) syn-syn bridging acetates. The X-ray diffraction analysis of 3 (C(40)H(46)N(14)O(8)Fe(4), monoclinic, P2(1)/c, a = 11.7633(18) A, b = 18.234(3) A, c = 10.4792(16) A, beta = 99.359(18) degrees, V = 2217.7(6) A(3), Z = 2) and 4 (C(34)H(46)N(26)O(2)Fe(4), monoclinic, P2(1)/c, V = 4412.4(10) A(3), a = 23.534(3) A, b = 18.046(2) A, c = 10.4865(16) A, beta = 97.80(2) degrees, Z = 4) showed a zigzag bis-dinuclear arrangement of four iron(II) cations, two mu(2)-O bridging pypentO ligands, four mu(2)-N-cyanato bridging ligands (3) or four end on azido bridging ligands (4): they are the first examples of cyanato and azido bridged discrete polynuclear ferrous compounds, respectively. The Mossbauer spectra of 1 are consistent with four different high-spin iron(II) sites in the Fe(4)O(4) cubane-type structure. The Mossbauer spectra of 3 are consistent with two high-spin iron(II) sites (N(5)O and N(4)O). Below 190 K, the Mossbauer spectra of 4 are consistent with one N(5)O and two N(4)O high-spin iron(II) sites. The temperature dependence of the magnetic susceptibility was fitted with J(1) approximately 0 cm(-1), J(2) = -1.3 cm(-1), J(3) = 4.6 cm(-1), D = 6.4 cm( 1), and g = 2.21 for 1; J(1) = 2.6 cm(-1), J(2) = 2.5 cm(-1), J(3) = - 5.6 cm( 1), D = 4.5 cm(-1), and g = 2.09 for 2; J(1) = 1.5 cm(-1), J(2) = 0.2 cm(-1), D = - 5.6 cm(-1), D' = 4.5 cm(-1), and g = 2.14 for 3; and J(1) = - 2.6 cm(-1), J(2) = 0.8 cm(-1), D= 6.3 cm(-1), D' = 1.6 cm(-1), and g = 2.18 for 4. The differences in sign among the J(1), J(2), and J(3) super-exchange interactions indicate that the faces including only mu(3)-OR bridges exhibit ferromagnetic interactions. The nature of the ground state in 1-3 is confirmed by simulation of the magnetization curves at 2 and 5 K. In the bis-dinuclear iron(II) compounds 3 and 4, the J(2) interaction resulting from the bridging of two Fe(2)(pypentO)X(3) units through two pseudo-halide anions is ferromagnetic in 3 (X = mu(2)-N-cyanato) and may be either ferro- or antiferromagnetic in 4 (X = end-on azido). The J(1) interaction through the central O(alkoxo) and pseudo-halide bridges inside the dinuclear units is ferromagnetic in 3 (X = mu(2)-N-cyanato) and antiferromagnetic in 4 (X = end-on azido). In agreement with the symmetry of the two Fe(II) sites in complexes 3 and 4, D (pentacoordinated sites) is larger than D' (octahedral sites). PMID- 11896718 TI - Formation of double cubanes [Sn(7)(NR)(8)] in the reactions of pyridyl and pyrimidinyl amines with Sn(NMe(2))(2): a synthetic and theoretical study. AB - In contrast to the reactions of Sn(NMe(2))(2) with unfunctionalized primary amines (RNH(2)), which yield the simple imido Sn(II) cubanes [SnNR](4), the reactions of 2-pyridyl or 2-pyrimidinyl amines give the mixed-oxidation-state Sn(II)/Sn(IV) double cubanes [Sn(7)(NR)(8)]. In addition to [Sn(7)[2-N(5 Mepy)](8)] x 2thf (1 x 2thf) (py = pyridine) and [Sn(7)[2-N(pm)](8)] x 0.33thf (2 x 0.33thf) (pm = pyrimidine), which were communicated previously, the syntheses and structures of the new complexes [Sn(7)[2-N(4-Mepm)](8)] x 2thf (3 x 2thf), [Sn(7)[2-N(4,6-Me(2)pm)](8)] x 4thf (4 x 4thf), [Sn(7)[2-N(4-Me-6-MeO-pm)](8)] (5), and [Sn(7)[2-N(4-MeO-6-MeO-pm)](8)] (6) are reported. Model DFT calculations on the reactions of Sn(NMe(2))(2) with 2-pmNH(2) or PhNH(2), producing the cubanes [Sn[2-N(pm)]](4) and [SnNPh](4) (respectively), and the corresponding double cubanes [Sn(7)[2-N(pm)](8)] and [Sn(7)(NPh)(8)], show that the presence of intramolecular Sn...N bonding which spans the cubane halves of the complexes is crucial to the formation of the double-cubane structure. PMID- 11896719 TI - Characteristics and properties of metal-to-ligand charge-transfer excited states in 2,3-bis(2-pyridyl)pyrazine and 2,2'-bypyridine ruthenium complexes. Perturbation-theory-based correlations of optical absorption and emission parameters with electrochemistry and thermal kinetics and related Ab initio calculations. AB - The absorption, emission, and infrared spectra, metal (Ru) and ligand (PP) half wave potentials, and ab initio calculations on the ligands (PP) are compared for several [L(n)()Ru(PP)](2+) and [[L(n)Ru]dpp[RuL'(n)]](4+) complexes, where L(n) and L'(n) = (bpy)(2) or (NH(3))(4) and PP = 2,2'-bipyridine (bpy), 2,3-bis(2 pyridyl)pyrazine (dpp), 2,3-bis(2-pyridyl)quinoxaline (dpq), or 2,3 bis(2pyridyl)benzoquinoxaline (dpb). The energy of the metal-to-ligand charge transfer (MLCT) absorption maximum (hnu(max)) varies in nearly direct proportion to the difference between Ru(III)/Ru(II) and (PP)/(PP)(-) half-wave potentials, DeltaE(1/2), for the monometallic complexes but not for the bimetallic complexes. The MLCT spectra of [(NH(3))(4)Ru(dpp)](2+) exhibit three prominent visible-near UV absorptions, compared to two for [(NH(3))(4)Ru(bpy)](2+), and are not easily reconciled with the MLCT spectra of [[(NH(3))(4)Ru]dpp[RuL(n)]](4+). The ab initio calculations indicate that the two lowest energy pi orbitals are not much different in energy in the PP ligands (they correlate with the degenerate pi orbitals of benzene) and that both contribute to the observed MLCT transitions. The LUMO energies calculated for the monometallic complexes correlate strongly with the observed hnu(max) (corrected for variations in metal contribution). The LUMO computed for dpp correlates with LUMO + 1 of pyrazine. This inversion of the order of the two lowest energy pi orbitals is unique to dpp in this series of ligands. Configurational mixing of the ground and MLCT excited states is treated as a small perturbation of the overall energies of the metal complexes, resulting in a contribution epsilon(s) to the ground-state energy. The fraction of charge delocalized, alpha(DA)(2), is expected to attenuate the reorganizational energy, chi(reorg), by a factor of approximately (1 - 4alpha(DA)(2) + alpha(DA)(4)), relative to the limit where there is no charge delocalization. This appears to be a substantial effect for these complexes (alpha(DA)(2) congruent with 0.1 for Ru(II)/bpy), and it leads to smaller reorganizational energies for emission than for absorption. Reorganizational energies are inferred from the bandwidths found in Gaussian analyses of the emission and/or absorption spectra. Exchange energies are estimated from the Stokes shifts combined with perturbation--theory-based relationship between the reorganizational energies for absorption and emission values. The results indicate that epsilon(s) is dominated by terms that contribute to electron delocalization between metal and PP ligand. This inference is supported by the large shifts in the N-H stretching frequency of coordinated NH(3) as the number of PP ligands is increased. The measured properties of the bpy and dpp ligands seem to be very similar, but electron delocalization appears to be slightly larger (10-40%) and the exchange energy contributions appear to be comparable (e.g., approximately 1.7 x 10(3) cm(-1) in [Ru(bpy)(2)dpp](2+) compared to approximately 1.3 x 10(3) cm(-1) in the bpy analogue). PMID- 11896720 TI - Structure and dynamics of the lincomycin-copper(II) complex in water solution by (1)H and (13)C NMR studies. AB - The copper(II) complex of lincomycin in water solution at pH = 7.15 was characterized by (1)H and (13)C NMR and UV-vis spectroscopy. A 1:1 complex is formed in these conditions. The temperature dependence of spin-lattice relaxation rates was measured, showing that all protons behave in a similar fashion and slow exchange conditions prevail. The spin-lattice relaxation rate enhancements were interpreted by the Solomon-Bloembergen-Morgan theory. Reorientational dynamics of the complex was approximated by evaluating the motional correlation time of free lincomycin in water solution. The observed proton and carbon relaxation rate enhancements allowed us to calculate copper-proton and copper-carbon distances that were used for building a molecular model of the complex. The obtained data provide an interpretation of the relatively high stability constant. PMID- 11896721 TI - Homologous series of redox-active, dinuclear cations [M(2)(O(2)CCH(3))(2)(pynp)(2)](2+) (M = Mo, Ru, Rh) with the bridging ligand 2-(2 pyridyl)-1,8-naphthyridine (pynp). AB - A homologous series of dinuclear compounds with the bridging ligand 2-(2-pyridyl) 1,8-naphthyridine (pynp) has been prepared and characterized by X-ray crystallographic and spectroscopic methods. [Mo(2)(O(2)CCH(3))(2)(pynp)(2)][BF(4)](2) x 3CH(3)CN (1) crystallizes in the monoclinic space group P2(1)/c with a = 15.134(5) A, b = 14.301(6) A, c = 19.990(6) A, beta = 108.06(2) degrees, V = 4113(3) A(3), and Z = 4. [Ru(2)(O(2)CCH(3))(2)(pynp)(2)][PF(6)](2) x 2CH(3)OH (2) crystallizes in the monoclinic space group C2/c with a = 14.2228(7) A, b = 20.3204(9) A, c = 14.1022(7) A, beta = 95.144(1) degrees, V = 4059.3(3) A(3), and Z = 4. [Rh(2)(O(2)CCH(3))(2)(pynp)(2)][BF(4)](2) x C(7)H(8) (3) crystallizes in the monoclinic space group C2/c with a = 13.409(2) A, b = 21.670(3) A, c = 13.726(2) A, beta = 94.865(2) degrees, V = 3973.9(8) A(3), and Z = 4. A minor product, [Rh(2)(O(2)CCH(3))(2)(pynp)(2)(CH(3)CN)(2)][BF(4)][PF(6)] x 2CH(3)CN (4), was isolated from the mother liquor after crystals of 3 had been harvested; this compound crystallizes in the triclinic space group, P1 with a = 12.535(3) A, b = 13.116(3) A, c = 13.785(3) A, alpha = 82.52(3) degrees, beta = 77.70(3) degrees, gamma = 85.76(3) degrees, V = 2193.0(8) A(3), and Z = 2. Compounds 1-3 constitute a convenient series for probing the influence of the electronic configuration on the extent of mixing of the M-M orbitals with the pi system of the pynp ligand. Single point energy calculations performed on 1-3 at the B3LYP level of theory lend insight into the bonding in these compounds and allow for correlations to be made with electronic spectral data. Although purely qualitative in nature, the values for normalized change in orbital energies (NCOE) of the frontier orbitals before and after reduction are in agreement with the observed differences in reduction potentials as determined by cyclic voltammetry. PMID- 11896722 TI - Ruthenium-manganese complexes for artificial photosynthesis: factors controlling intramolecular electron transfer and excited-state quenching reactions. AB - Continuing our work toward a system mimicking the electron-transfer steps from manganese to P(680)(+) in photosystem II (PS II), we report a series of ruthenium(II)-manganese(II) complexes that display intramolecular electron transfer from manganese(II) to photooxidized ruthenium(III). The electron transfer rate constant (k(ET)) values span a large range, 1 x 10(5)-2 x 10(7) s( 1), and we have investigated different factors that are responsible for the variation. The reorganization energies determined experimentally (lambda = 1.5 2.0 eV) are larger than expected for solvent reorganization in complexes of similar size in polar solvents (typically lambda approximately 1.0 eV). This result indicates that the inner reorganization energy is relatively large and, consequently, that at moderate driving force values manganese complexes are not fast donors. Both the type of manganese ligand and the link between the two metals are shown to be of great importance to the electron-transfer rate. In contrast, we show that the quenching of the excited state of the ruthenium(II) moiety by manganese(II) in this series of complexes mainly depends on the distance between the metals. However, by synthetically modifying the sensitizer so that the lowest metal-to-ligand charge transfer state was localized on the nonbridging ruthenium(II) ligands, we could reduce the quenching rate constant in one complex by a factor of 700 without changing the bridging ligand. Still, the manganese(II)-ruthenium(III) electron-transfer rate constant was not reduced. Consequently, the modification resulted in a complex with very favorable properties. PMID- 11896723 TI - Chiral monomeric and homochiral dimeric copper(II) complexes of a new chiral ligand, N-(1,2-bis(2-pyridyl)ethyl)pyridine-2-carboxamide: an example of molecular self-recognition. AB - Reaction of Cu(ClO(4))(2) x 6H(2)O with a racemic mixture of the novel chiral ligand N-(1,2-bis(2-pyridyl)ethyl)pyridine-2-carboxamide (PEAH) affords only the homochiral dimeric copper(II) complexes [Cu(2)((R)()PEA)(2)](ClO(4))(2) and [Cu(2)((S)()PEA)(2)](ClO(4))(2) in a 1:1 ratio. The phenomenon of molecular self recognition is also observed when a racemic mixture of the monomeric copper(II) complex [Cu((R(S))()PEA)(Cl)(H(2)O)] is converted into the homochiral dimeric species [Cu(2)((R(S))()PEA)(2)](ClO(4))(2) via reaction with Ag(+) ion. This is the first report of direct conversion of a racemic mixture of a chiral monomeric copper(II) complex to a mixture of the homochiral dimers. PMID- 11896725 TI - Anisole hydrogenation with well-characterized polyoxoanion- and tetrabutylammonium-stabilized Rh(0) nanoclusters: effects of added water and acid, plus enhanced catalytic rate, lifetime, and partial hydrogenation selectivity. AB - Following a comprehensive look at the arene hydrogenation literature by soluble nanocluster catalysts, six key, unfulfilled goals in nanocluster arene hydrogenation catalysis are identified. To begin to address those six goals, well characterized polyoxoanion- and tetrabutylammonium-stabilized Rh(0) nanoclusters have been synthesized by the reduction of the precisely defined precatalyst [Bu(4)N](5)Na(3)[(1,5-COD)Rh small middle dotP(2)W(15)Nb(3)O(62)] with H(2) in propylene carbonate solvent. These Rh(0) nanoclusters are characterized by their stoichiometry of formation, transmission electron microscopy, and the two rate constants which characterize their mechanism of formation; previous studies in our laboratories have provided additional characterization of polyoxoanion stabilized Rh(0) nanoclusters. Propylene carbonate solutions of the Rh(0) nanoclusters catalyze the hydrogenation of anisole (methoxybenzene) under mild conditions (22-78 degrees C, 30-40 psig H(2)). Proton donors such as water or HBF(4) small middle dotEt(2)O are discovered to affect both nanocluster formation and nanocluster arene hydrogenation catalysis. Under identical conditions, the Rh(0) nanoclusters are 10-fold more active than a commercially available, oxide supported 5% Rh/Al(2)O(3) catalyst of the same average metal-particle size. A series of lifetime experiments shows that the Rh(0) nanoclusters are capable of at least 2600 total turnovers (TTO), a lifetime significantly longer than the approximately 100 TTO often seen for nanocluster arene hydrogenation catalysts, and a lifetime slightly better than the prior record of 2000 TTO for a literature nanocluster system. The present polyoxoanion-stabilized Rh(0) nanoclusters also display a record, albeit modest, 30% selectivity for the partial hydrogenation of anisole to 1-methoxycyclohexene with an overall yield of up to 8% at higher temperatures. In comparison to the 5% Rh/Al(2)O(3) catalyst, the polyoxoanion stabilized nanoclusters yield a 4.7-fold higher maximum yield of 1 methoxycyclohexene. Finally, the seven main findings of the present work are summarized, including how they address five of the aforementioned six main goals in nanocluster arene hydrogenation. PMID- 11896724 TI - Metal-induced cyclization of thiosemicarbazones derived from beta-keto amides and beta-keto esters: open-chain and cyclized ligands in zinc(II) complexes. AB - The reactions of Zn(OAc)(2) with acetoacetanilide, methyl acetoacetate, o acetoacetanisidide, and ethyl 2-methylacetoacetate thiosemicarbazones (HTSC(1), HTSC(2), HTSC(3), and HTSC(4), respectively) were explored in methanol. With HTSC(1), HTSC(2), and HTSC(3), following isolation of the corresponding zinc(II) thiosemicarbazonates [Zn(TSC(x))(2)] (x = 1, 2, 3), the mother liquors afforded pyrazolonate complexes [ZnL(1)(2)(H(2)O)] (HL(1) = 2,5-dihydro-3-methyl-5-oxo-1H pyrazole-1-carbothioamide) that had been formed by cyclization of the corresponding TSC(-). The reaction of HTSC(4) with zinc(II) acetate gave only the pyrazolonate complex [ZnL(2)(2)(H(2)O)] (HL(2) = 2,5-dihydro-3,4-dimethyl-5-oxo 1H-pyrazole-1-carbothioamide). All compounds were studied by IR and NMR spectroscopy, and HTSC(3), [Zn(TSC(3))(2)] x DMSO, [ZnL(1)(2)(H(2)O)] x 2DMSO, and [ZnL(2)(2)(H(2)O)] x 2DMSO were also studied by X-ray diffractometry, giving a thorough picture of the cyclization process. In preliminary tests of the effects of HL(1) and [ZnL(1)(2)(H(2)O)] on rat paw inflammatory edema induced by carrageenan, HL(1) showed antiinflammatory activity. PMID- 11896726 TI - Quantifying the electronic effect of substituted phosphine ligands via molecular electrostatic potential. AB - Values of the molecular electrostatic potential minimum (V(min)) corresponding to the lone pair region of several substituted phosphine ligands (PR(3)) have been determined at the DFT level. The V(min) value is proposed as a quantitative measure of the electronic effect of the PR(3) ligands. Good linear correlation between V(min) and Tolman electronic parameter of PR(3) has been obtained. V(min) is also proportional to the pK(a) values of the conjugate acids of PR(3), viz., [PR(3)H](+). Further, the DeltaE values of the reaction Ni(CO)(3) + PR(3) --> Ni(CO)(3)PR(3) and ScH(3) + PR(3) --> ScH(3)PR(3) are also linearly proportional to the V(min) values. However, if there is a strong metal to phosphorus pi-back bonding, the DeltaE and V(min) do not fit to a line. It is also found that the standard reduction potential as well as the enthalpy change corresponding to the electrochemical couple eta-Cp(CO)(PR(3))(COMe)Fe(+)/eta-Cp(CO)(PR(3))(COMe)Fe(0) is linearly proportional to the V(min) values of PR(3). These correlations suggest that V(min) is a quantitative measure of the sigma-donating ability of the phosphine. It is hoped that, in phosphine-metal coordination chemistry, the V(min) based electronic parameter could be more advantageous than nu-CO and pK(a) based electronic parameters as it solely represents the inherent electronic property of the ligand. PMID- 11896727 TI - Mechanistic information from pressure acceleration of hydride formation via proton binding to a cobalt(I) macrocycle. AB - The effect of pressure on proton binding to the racemic isomer of the cobalt(I) macrocycle, CoL(+) (L = 5,7,7,12,14,14-hexamethyl-1,4,8,11-tetraazacyclotetradeca 4,11-diene), has been studied for a series of proton donors using pulse radiolysis techniques. The second-order rate constants for the reaction of CoL(+) with proton donors decrease with increasing pK(a) of the donor acid, consistent with a reaction occurring via proton transfer. Whereas the corresponding volumes of activation (DeltaV) are rather small and negative for all acids (proton donors) with pK(a) values below 8.5, significantly larger negative activation volumes are found for weaker acids (pK(a) > 9.5) containing OH groups as proton donors. In the latter case, the observed DeltaV for these protonation reactions show a correlation with the reaction volumes (DeltaV degrees (ion)) for the ionization of the weak acids with a slope of 0.44, indicating that bond dissociation of the weak acid molecule bound to the metal center proceeds approximately halfway at the transition state along the reaction coordinate in terms of volume changes. PMID- 11896729 TI - Chelated hydrazido(3-)rhenium(V) complexes: on the way to the nitrido-M(V) core (M = Tc, Re). AB - Neutral and asymmetrical hydrazido(3-)rhenium(V) heterocomplexes of the type [Re(eta(2)-L(4))(L(n))(PPh(3))] (eta(2)-L(4) = NNC(SCH(3))S; H(2)L(1) = S-methyl beta-N-((2-hydroxyphenyl)ethylidene)dithiocarbazate, 1, H(2)L(2) = S-methyl beta N-((2-hydroxyphenyl)methylidene)dithiocarbazate, 2) are prepared via ligand exchange reactions in ethanolic solutions starting from [Re(V)(O)Cl(4)](-) in the presence of PPh(3) or from [Re(V)(O)Cl(3)(PPh(3))(2)]. The distorted octahedral coordination sphere of these compounds is saturated by a chelated hydrazido group, a facially ligated ONS Schiff base, and PPh(3). Reduction-substitution reactions starting from [NH(4)][Re(VII)O(4)] in acidic ethanolic mixtures containing PPh(3) and H(2)L(n) (or its dithiocarbazic acid precursor H(3)L(4)) produce another example of chelated hydrazido(3-) rhenium(V) derivative, namely [Re(eta(2)-L(4))Cl(2)(PPh(3))(2)], 3. On the contrary, the N-methyl-substituted dithiocarbazic acid H(2)L(3) reacts with perrhenate to give the known nitrido complex [Re(N)Cl(2)(PPh(3))(2)]. Rhenium(V) complexes incorporating the robust eta(2)-hydrazido moiety represent key intermediates helpful for the comprehension of the reaction pathway which generates nitridorhenium(V) species starting from oxo precursors. An essential requirement for the stabilization of such chelated hydrazido-Re(V) units is the triple deprotonation at the hydrazine nitrogens, thereby providing efficient pi-electron circulation in the resulting five membered ring. The thermal stability of these units is affected by the nature of the anchoring donor, thione sulfur ensuring stronger chelation than nitrogen and oxygen. The eta(2)-hydrazido complexes are characterized by conventional physicochemical techniques, including the X-ray crystal structure determination of 1 and 3. PMID- 11896728 TI - Thermodynamic studies on the recognition of flexible peptides by transition-metal complexes. AB - Strong and selective binding to a trihistidine peptide has been achieved employing Cu(2+)-histidine interactions in aqueous medium (25 mM HEPES buffer, pH 7.0). When the pattern of cupric ions on a complex matched with the pattern of histidines on the peptide, a strong and selective binding was observed. UV-vis spectroscopic studies show that the cupric ions coordinate to the histidines of the peptides. Thermodynamic studies reveal that the binding process is enthalpy driven over the entire range of working temperature (25-40 degrees C). An enthalpy-entropy compensation effect was also observed. PMID- 11896730 TI - Synthesis, structures, and magnetic properties of heterodimetal bis(mu hydroxo)chromium(III)nickel(II) complexes with Tpa derivatives having sterically bulky substituents. AB - A series of heterodinuclear bis(mu-hydroxo)chromium(III)nickel(II) complexes was newly prepared: [(phen)(2)Cr(mu-OH)(2)Ni(tpa)](ClO(4))(3) x 0.5H(2)O (1), [(phen)(2)Cr(mu-OH)(2)Ni(Me-tpa)](ClO(4))(3) x 2H(2)O (2), [(phen)(2)Cr(mu OH)(2)Ni(Me(2)-tpa)](ClO(4))(3) x 2H(2)O (3), and [(phen)(2)Cr(mu-OH)(2)Ni(Me(3) tpa)](ClO(4))(3) x 3H(2)O (4), where phen is 1,10-phenanthroline and tpa, Me-tpa, Me(2)-tpa, and Me(3)-tpa are tris(2-pyridylmethyl)amine, [(6-methyl-2 pyridyl)methyl]bis(2-pyridylmethyl)amine, bis[(6-methyl-2-pyridyl)methyl](2 pyridylmethyl)amine, and tris[(6-methyl-2-pyridyl)methyl]amine, respectively. X ray crystallography revealed that the structures of 1-4 resemble one another having an edge-shared bioctahedral structure with a Cr(mu -OH)(2)Ni unit (crystal data: 1 x C(2)H(5)OH, triclinic, P1, a = 13.179(4) A, b = 13.685(4) A, c = 14.260(4) A, alpha = 84.95(2) degrees, beta = 77.65(1) degrees, gamma = 90.21(2) degrees, V = 2502(1) A(3), Z = 2, R = 0.103, R(w) = 0.097; 2 x C(2)H(5)OH, triclinic, P1, a = 13.214(2) A, b = 13.657(2) A, c = 14.417(3) A, alpha = 95.205(5) degrees, beta = 102.583(4) degrees, gamma =90.720(3) degrees, V = 2527.3(8) A(3), Z = 2, R = 0.090, R(w) = 0.122; 3 x C(2)H(5)OH, triclinic, P1, a = 13.276(2) A, b =13.696(2) A, c = 14.454(2) A, alpha = 95.640(3) degrees, beta = 102.821(4) degrees, gamma = 90.174(3) degrees, V = 2549.5(6) A(3), Z = 2, R= 0.087, R(w)= 0.119; 4, triclinic, P1, a = 10.8916(9) A, b = 14.268(2) A, c = 17.522(2) A, alpha = 84.498(9) degrees, beta = 74.313(7) degrees, gamma = 72.402(7) degrees, V = 2498.6(5) A(3), Z = 2, R = 0.060, R(w)= 0.088). Chromium and nickel ions are coordinated by two phen's and Me(n)-tpa, respectively, to complete a distorted octahedral coordination sphere. Introduction of the 6-methyl group(s) onto the pyridyl group(s) results in the elongation of the Ni-N bond distances due to an unfavorable steric interaction between the methyl group and the bridging hydroxide group: systematic elongation of the Ni-N bond distances and the Cr ...Ni separations accompanied by an increase in the Cr-O-Ni angles was observed as the number of the methyl groups increases. Variable-temperature magnetic susceptibility measurements of 1-4 (4.2-300 K) indicated that magnetic interactions between Cr(III) and Ni(II) ions are systematically modulated from a very weak antiferromagnetic interaction to a ferromagnetic interaction as the number of the methyl groups increases; the exchange integrals J's for 1-4 are estimated to be -1.4, +0.0, +4.1, and +7.4 cm(-1), respectively. The magneto structural relationship is discussed in terms of the change in the magnetic orbital energies of nickel(II) centers arising from the change in the Ni-N bond distances. PMID- 11896731 TI - In situ formation of ruthenium catalysts for the homogeneous hydrogenation of carbon dioxide. AB - A total of 44 different phosphines were tested, in combination with [RuCl(2)(C(6)H(6))](2) and three other Ru(II) precursors, for their ability to form active catalysts for the hydrogenation of CO(2) to formic acid. Half (22) of the ligands formed catalysts of significant activity, and only 6 resulted in very high rates of production of formic acid. These were PMe(3), PPhMe(2), dppm, dppe, and cis- and trans-Ph(2)PCH=CHPPh(2). The in situ catalysts prepared from [RuCl(2)(C(6)H(6))](2) and any of these 6 phosphine ligands were found to be at least as efficient as the isolated catalyst RuCl(O(2)CMe)(PMe(3))(4). There was no correlation between the basicity of monophosphines (PR(3)) and the activity of the catalysts formed from them. However, weakly basic diphosphines formed highly active catalysts only if their bite angles were small, while more strongly basic diphosphines had the opposite trend. In situ (31)P NMR spectroscopy showed that trans-Ru(H)(2)(dppm)(2), trans-RuCl(2)(dppm)(2), trans-RuHCl(dppm)(2), cis Ru(H)(O(2)CH)(dppm)(2), and cis-Ru(O(2)CH)(2)(dppm)(2) are produced as the major metal-containing species in reactions of dppm with [RuCl(2)(C(6)H(6))](2) under catalytic conditions at 50 degrees C. PMID- 11896732 TI - Single- and double-cubane clusters in the multiple oxidation states [VFe(3)S(4)](3+,2+,1+). AB - A new series of cubane-type [VFe(3)S(4)](z)() clusters (z = 1+, 2+, 3+) has been prepared as possible precursor species for clusters related to those present in vanadium-containing nitrogenase. Treatment of [(HBpz(3))VFe(3)S(4)Cl(3)](2)(-) (2, z = 2+), protected from further reaction at the vanadium site by the tris(pyrazolyl)hydroborate ligand, with ferrocenium ion affords the oxidized cluster [(HBpz(3))VFe(3)S(4)Cl(3)](1)(-) (3, z = 3+). Reaction of 2 with Et(3)P results in chloride substitution to give [(HBpz(3))VFe(3)S(4)(PEt(3))(3)](1+) (4, z = 2+). Reaction of 4 with cobaltocene reduced the cluster with formation of the edge-bridged double-cubane [(HBpz(3))(2)V(2)Fe(6)S(8)(PEt(3))(4)] (5, z = 1+, 1+), which with excess chloride underwent ligand substitution to afford [(HBpz(3))(2)V(2)Fe(6)S(8)Cl(4)](4)(-) (6, z = 1+, 1+). X-ray structures of (Me(4)N)[3], [4](PF(6)), 5, and (Et(4)N)(4)[6] x 2MeCN are described. Cluster 5 is isostructural with previously reported [(Cl(4)cat)(2)(Et(3)P)(2)Mo(2)Fe(6)S(8)(PEt(3))(4)] and contains two VFe(3)S(4) cubanes connected across edges by a Fe(2)S(2) rhomb in which the bridging Fe-S distances are shorter than intracubane Fe-S distances. Mossbauer (2-5), magnetic (2-5), and EPR (2, 4) data are reported and demonstrate an S = 3/2 ground state for 2 and 4 and a diamagnetic ground state for 3. Analysis of (57)Fe isomer shifts based on an empirical correlation between shift and oxidation state and appropriate reference shifts results in two conclusions. (i) The oxidation 2 --> 3 + e(-) results in a change in electron density localized largely or completely on the Fe(3) subcluster and associated sulfur atoms. (ii) The most appropriate charge distributions are [V(3+)Fe(3+)Fe(2+)(2)S(4)](2+) (Fe(2.33+)) for 1, 2, and 4 and [V(3+)Fe(3+)(2)Fe(2+)S(4)](3+) (Fe(2.67+)) for 3 and [V(2)Fe(6)S(8)(SEt)(9)](3+). Conclusion i applies to every MFe(3)S(4) cubane-type cluster thus far examined in different redox states at parity of cluster ligation. The formalistic charge distributions are regarded as the best current approximations to electron distributions in these delocalized species. The isomer shifts require that iron atoms are mixed-valence in each cluster. PMID- 11896733 TI - Nanoclusters in catalysis: a comparison of CS(2) catalyst poisoning of polyoxoanion- and tetrabutylammonium-stabilized 40 +/- 6 A Rh(0) nanoclusters to 5 Rh/Al(2)O(3), including an analysis of the literature related to the CS(2) to metal stoichiometry issue. AB - It is crucial in metal particle catalysis to know the true number of catalytically active surface sites; without this knowledge it is impossible (i) to know the true turnover frequency (TOF, i.e., the moles of product/(moles of active metal atoms x time)); (ii) to know for certain whether a (quantitatively) better catalyst has been made-on a per-active-metal-atom basis; (iii) to know the amount of active sites remaining in a deactivated catalyst; and (iv) to know how many active sites have been regenerated in a reactivated catalyst. For this reason, herein we report the first quantitative, more complete and fundamental study of nanocluster catalyst poisoning using the preferred CS(2) method with polyoxoanion- and tetrabutylammonium-stabilized Rh(0) nanoclusters; 5% Rh/Al(2)O(3) is also examined as a valuable comparison point. Both catalysts are examined under essentially identical conditions and while catalyzing a prototype reaction, cyclohexene hydrogenation. A number of control studies are also reported to be sure that the kinetic method used to follow the CS(2) poisoned hydrogenation reaction is reliable, to test for H(2) gas-to-solution mass transfer limitations, to test for reversibility in the CS(2) poisoning, and to test for loss of the volatile CS(2). The results allow 10 previously unavailable insights and conclusions, including the first quantitative comparison of the active-site corrected TOF for a nanocluster catalyst (in this case Rh(0) nanoclusters) to its supported heterogeneous counterpart (the 5% Rh(0) on Al(2)O(3)). The results show that the nanocluster surface Rh(0) is between 2.3 and 23 times more active on a per-active-metal-atom basis. Overall, the results introduce to the transition-metal nanocluster area the catalyst poisoning methodology necessary for the determination of the number of active metal sites. The important literature of CS(2) catalyst poisoning studies is also cited and discussed with a focus on the previously neglected issue of the exact poison/metal stoichiometry ratio. Significantly, the single metal crystal plus CS(2) literature provides evidence that the CS(2)/metal ratio probably lies between 1/1.5 and 1/10 in most cases. The data presented herein suggest that the CS(2)/Rh ratio for the Rh(0) nanoclusters is very likely within this range and for certain is <1/17. PMID- 11896734 TI - Filling a void: isolation and characterization of tetracarboxylato dimolybdenum cations. AB - Dimolybdenum tetracarboxylato cations have been prepared and structurally characterized for the first time. The reactions of the new, quadruply bonded compound, Mo(2)(TiPB)(4), where TiPB = 2,4,6-triisopropylphenyl carboxylate, with NOPF(6) and NOBF(4) give the ionic compounds [Mo(2)(TiPB)(4)]PF(6) and [Mo(2)(TiPB)(4)]BF(4), respectively. Each product crystallizes in space group P2(1)/n and displays an elongation of the Mo-Mo bond of 0.060 and 0.068 A, respectively, over that of the parent compound (2.076(1) A). Each complex displays a characteristic EPR signal, showing hyperfine coupling to the spin active isotopes (95)Mo and (97)Mo, with g( parallel) = g( perpendicular) = 1.936, that is consistent with the presence of an unpaired electron. Electronic spectroscopy indicates the expected red shift in the delta --> delta(*) transition for the cations, due to the loss of exchange energy in going from the two-electron to one-electron system. We have also obtained a small amount of crystalline [Mo(2)(O(2)CC(4)H(9))(4)]PF(6) from the reaction of Mo(2)(O(2)CC(4)H(9))(4) with AgPF(6). This product crystallizes in the space group C2/c, and the Mo-Mo bond is elongated by 0.063 A over that of the neutral parent compound. These ionic compounds have the first isolated and characterized [Mo(2)(O(2)CR)(4)](+) cationic species. PMID- 11896736 TI - Structures of Sb(OC(6)H(3)Me(2)-2,6)(3) and Sb(OEt)(5) x NH(3): the first authenticated monomeric Sb(OR)(n) (n = 3, 5). AB - The first monomeric antimony alkoxides, Sb(OC(6)H(3)Me(2))(3) (1) and Sb(OEt)(5) x NH(3) (2), have been crystallographically characterized. The former adopts a trigonal pyramidal geometry, while the latter is octahedral about antimony; hydrogen bonding between NH(3) and SbOEt groups in Sb(OEt)(5) small middle dotNH(3) creates a one-dimensional lattice arrangement. Reaction of pyridine with SbCl(5) in EtOH/hexane yields the salt [Hpy(+)](9)[Sb(2)Cl(11)(5)(-)][Cl(-)](4) (3), which has also been crystallographically characterized. Crystallographic data: 1, C(24)H(27)O(3)Sb, a = 10.9080(2), b = 11.9660(2), c = 17.7260(4) A, alpha = 109.740(1) degrees, monoclinic P2(1)/c (unique axis a), Z = 4; 2, C(10)H(28)NO(5)Sb, a = 7.7220(1), b = 19.0700(2), c = 21.6800(3) A, beta = 93.4960(7) degrees, monoclinic P2(1)/c, Z = 8; 3, C(45)H(54)Cl(15)N(9)Sb(2), a = 13.4300(2), b = 14.4180(2), c = 17.4180(3) A, alpha = 82.7650(7), beta = 77.5570(7), gamma = 70.7670(7) degrees, triclinic P1, Z = 2. PMID- 11896735 TI - P-O donor action from carboxylate anions with phosphorus in the presence of hydrogen bonding. A model for phosphoryl-transfer enzymes. AB - A series of phosphorus compounds (1-3) containing anionic carboxylate groups were synthesized by treatment of the respective neutral precursor acid forms B-D with amines, which also served to introduce hydrogen-bonding interactions. The compounds, subjected to X-ray structure analysis, resulted in hexacoordinated anionic phosphoranates 1A and 1B, a pseudo-trigonal-bipyramidal anionic phosphine (2), and a trigonal-bipyramidal anionic phosphine oxide (3). The structures revealed that P-O donor coordination was present in all members of the anionic series 1-3 and resulted in stronger interactions than existed in the precursor neutral acid forms B-D as measured by the presence of shorter P-O distances. Evaluation of the energies of the donor interactions relative to the energies of the hydrogen bonds that were present showed that the donor energies now exceeded the hydrogen bond strengths. (31)P chemical shifts indicated that the basic coordination geometries were retained in solution. Both 1A and 1B are chiral and exist as racemates. The results suggest that mechanisms of phosphoryl-transfer enzymes should benefit by taking into account donor interactions at phosphorus by residues at active sites in addition to the inclusion of hydrogen bonding. Reference is made to specific phosphoryl-transfer enzymes. PMID- 11896737 TI - Insertion of a bis(phosphine)platinum group into the S-S bond of Mn(2)(CO)(7)(mu S(2)). AB - The reaction of Mn(2)(CO)(7)(mu-S(2)), 1, with Pt(PPh(3))(2)(PhC(2)Ph) yielded the new complex, Mn(2)(CO)(6)Pt(PPh(3))(2)(mu(3)-S)(2), 3, by loss of CO and insertion of a Pt(PPh(3))(2) group into the S-S bond of 1. Complex 3 was characterized crystallographically and was found to consist of an open Mn(2)Pt cluster with one Mn-Mn bond, 2.8154(14) A, one Mn-Pt bond, 2.9109(10) A, and two triply bridging sulfido ligands. Compound 3 reacts with CO to form adduct Mn(2)(CO)(6)(mu-CO)Pt(PPh(3))(2)(mu(3)-S)(2), 4. Compound 4 also contains an open Mn(2)Pt cluster with two triply bridging sulfido ligands but has only one metal metal bond, Mn-Mn = 2.638(2) A. Under nitrogen, compound 4 readily loses CO and reverts back to 3. PMID- 11896738 TI - Rhenium bipyridine complexes for the recognition of glucose. AB - Bipyridine ligands containing pendant methyl, amino, and amino-boronic acid groups were synthesized. Coordination complexes of these ligands with rhenium were prepared straightforwardly and in good yield. The fluorescence behavior of the Re complexes was studied as a function of pH and exposure to various concentrations of glucose. The methyl bipyridine complex showed no change in fluorescence with pH, the amino derivative showed a rapid decrease from low pH to neutral, and the amino-boronate derivative showed little change from pH 4 to 10. Fluorescence quenching was observed at high pH as expected on the basis of a photoinduced electron transfer (PET) signaling mechanism. This behavior can be explained on the basis of the first oxidation and reduction potentials of these complexes. Glucose testing showed a significant dependence on the solvent system used. In pure methanol, the rhenium boronate complex exhibited a 55% fluorescence intensity increase upon increasing glucose concentration from 0 to 400 mg/dL. However, in 50 vol % methanol/phosphate buffered saline, none of the complexes showed significant response in the glucose range of physiological interest. PMID- 11896739 TI - Preparative and structural studies on the carbonyl cyanides of iron, manganese, and ruthenium: fundamentals relevant to the hydrogenases. AB - The reaction of cyanide, carbon monoxide, and ferrous derivatives led to the isolation of three products, trans- and cis-[Fe(CN)(4)(CO)(2)](2)(-) and [Fe(CN)(5)(CO)](3)(-), the first two of which were characterized by single crystal X-ray diffraction. The new compounds show self-consistent IR, (13)C NMR, and mass spectroscopic properties. The reaction of trans-[Fe(CN)(4)(CO)(2)](2)(-) with Et(4)NCN gives [Fe(CN)(5)(CO)](3)(-) via a first-order (dissociative) pathway. The corresponding cyanation of cis-[Fe(CN)(4)(CO)(2)](2)(-), which is a minor product of the Fe(II)/CN(-)/CO reaction, does not proceed at measurable rates. Methylation of [Fe(CN)(5)(CO)](3)(-) gave exclusively cis [Fe(CN)(4)(CNMe)(CO)](2)(-), demonstrating the enhanced nucleophilicity of CN(-) trans to CN(-) vs. CN(-) trans to CO. Methylation has an electronic effect similar to that of protonation as determined electrochemically. We also characterized [M(CN)(3)(CO)(3)](n)(-) for Ru (n = 1) and Mn (n = 2) derivatives. The Ru complex, which is new, was prepared by cyanation of a [RuCl(2)(CO)(3)](2) solution. PMID- 11896740 TI - Redox-related chemical shift perturbations on backbone nuclei of high-potential iron-sulfur proteins. PMID- 11896741 TI - Valence delocalization in a mixed-oxidation divanadium (IV, V) complex electrogenerated from its structurally characterized divanadium (V) analogue with a tridentate (ONO) ligand. PMID- 11896743 TI - Dietary polyunsaturated fatty acids: impact on cancer chemotherapy and radiation. AB - Preclinical studies have shown that certain polyunsaturated fatty acids may actually enhance the cytotoxicity of several antineoplastic agents and the anticancer effects of radiotherapy. These effects are possibly mediated by incorporation of the polyunsaturated fatty acids into cancer cell membranes, thus altering the physical and functional properties. In addition, certain polyunsaturated fatty acids may also reduce or prevent some of the side effects of these therapies, and administering antioxidants to prevent polyunsaturated fatty acid-induced oxidative stress may further enhance the impact of chemotherapy and radiation. PMID- 11896744 TI - Sulfur in human nutrition and applications in medicine. AB - Because the role of elemental sulfur in human nutrition has not been studied extensively, it is the purpose of this article to emphasize the importance of this element in humans and discuss the therapeutic applications of sulfur compounds in medicine. Sulfur is the sixth most abundant macromineral in breast milk and the third most abundant mineral based on percentage of total body weight. The sulfur-containing amino acids (SAAs) are methionine, cysteine, cystine, homocysteine, homocystine, and taurine. Dietary SAA analysis and protein supplementation may be indicated for vegan athletes, children, or patients with HIV, because of an increased risk for SAA deficiency in these groups. Methylsulfonylmethane (MSM), a volatile component in the sulfur cycle, is another source of sulfur found in the human diet. Increases in serum sulfate may explain some of the therapeutic effects of MSM, DMSO, and glucosamine sulfate. Organic sulfur, as SAAs, can be used to increase synthesis of S-adenosylmethionine (SAMe), glutathione (GSH), taurine, and N-acetylcysteine (NAC). MSM may be effective for the treatment of allergy, pain syndromes, athletic injuries, and bladder disorders. Other sulfur compounds such as SAMe, dimethylsulfoxide (DMSO), taurine, glucosamine or chondroitin sulfate, and reduced glutathione may also have clinical applications in the treatment of a number of conditions such as depression, fibromyalgia, arthritis, interstitial cystitis, athletic injuries, congestive heart failure, diabetes, cancer, and AIDS. Dosages, mechanisms of action, and rationales for use are discussed. The low toxicological profiles of these sulfur compounds, combined with promising therapeutic effects, warrant continued human clinical trails. PMID- 11896745 TI - Alternative therapies for type 2 diabetes. AB - Type 2 diabetes is a chronic metabolic disease that has a significant impact on the health, quality of life, and life expectancy of patients, as well as on the health care system. Exercise, diet, and weight control continue to be essential and effective means of improving glucose homeostasis. However, lifestyle management measures may be insufficient or patient compliance difficult, rendering conventional drug therapies (i.e., oral glucose-lowering agents and insulin injection) necessary in many patients. In addition to adverse effects, drug treatments are not always satisfactory in maintaining euglycemia and avoiding late stage diabetic complications. As an alternative approach, medicinal herbs with antihyperglycemic activities are increasingly sought by diabetic patients and health care professionals. Commonly used herbs and other alternative therapies, less likely to have the side effects of conventional approaches for type 2 diabetes, are reviewed. PMID- 11896746 TI - Effective treatment of seborrheic dermatitis using a low dose, oral homeopathic medication consisting of potassium bromide, sodium bromide, nickel sulfate, and sodium chloride in a double-blind, placebo-controlled study. AB - BACKGROUND: Topical over-the-counter remedies exist to aid in the control of seborrheic dermatitis and chronic dandruff on a superficial level. Low-dose systemic oral nickel and bromide therapy has shown promise in providing improvement and eventual clearing of the disease. OBJECTIVE: The purpose of this study was to further evaluate the effect of an orally administered low-dose, homeopathic mineral therapy (Potassium bromide 1X, Sodium bromide 2X, Nickel sulfate 3X, Sodium chloride 6X) on seborrheic dermatitis and chronic dandruff. METHODS: Forty-one patients with seborrheic dermatitis and/or chronic dandruff were assigned to one of two treatment groups: Active (containing the medication) or placebo (vehicle). Study medication was administered in a placebo-controlled, randomly-selected, double-blind study for 10 weeks. At the end of 10 weeks all patients crossed over to the active medication, under a different label for an additional 10 weeks in an open study format. RESULTS: Twenty-nine patients completed the 10-week blinded portion of the study. After 10 weeks of treatment, the disease state of the active patients improved significantly over that of the placebo patients (p<0.04). The placebo patients' condition before and after crossover to active treatment was also evaluated, showing significant improvement (p<0.01) 10 weeks after crossing over to active medication. CONCLUSION: Oral therapy using a low-dose homeopathic preparation combining Potassium bromide 1X, Sodium bromide 2X, Nickel sulfate 3X, and Sodium chloride 6X, provides significant improvement in seborrheic dermatitis and dandruff after 10 weeks of dosing. PMID- 11896747 TI - Undecylenic acid. Monograph. PMID- 11896748 TI - Pygeum africanum (Prunus africanus) (African plum tree). Monograph. PMID- 11896749 TI - The clinical significance of recognizing distinct morphologic and biologic features of hereditary breast cancer. PMID- 11896750 TI - International consensus conference. Image-detected breast cancer: state of the art diagnosis and treatment. PMID- 11896751 TI - Acceleration of chromosomal instability of BRCA1-associated hereditary breast cancers by p53 abnormality. AB - Chromosomal instability (CIN) of BRCA1-associated hereditary breast cancers was studied by fluorescence in situ hybridization of chromosomes 1, 11, and 17. CIN values (the percentage of cells with a nonmodal chromosome) were obtained for each of three chromosomes, and their average was finally used as the CIN value of the sample. BRCA1-associated tumors showed a significantly (p = 0.0089) higher CIN value than normal breast tissues (24.3 +/- 4.3, n = 7, vs. 9.2 +/- 1.1, n = 6, mean +/- SE). In addition, BRCA1-associated tumors with positive p53 immunostaining showed a significantly (p = 0.0018) higher CIN value than those with negative p53 immunostaining (35.7 +/- 3.5, n = 3, vs. 15.8 +/- 1.3, n = 4). These results demonstrate that a loss of BRCA1 function results in the growth of breast cancers with CIN, and this phenotype is further accelerated by p53 abnormality. PMID- 11896752 TI - Local-regional breast cancer recurrence: prognostic groups based on patterns of failure. AB - The purpose of this study was to determine the outcome of breast cancer patients sustaining local-regional failure as their first site of relapse in an effort to group patients into prognostic categories. Between January 1970 and December 1992, over 4,000 patients with breast cancer were treated at our facilities with mastectomy or conservative surgery with radiation therapy (CS + RT). Two hundred thirteen patients sustained local-regional relapse without evidence of distant metastasis as their first site of failure, and they served as the population base for this study. The 213 patients with local-regional recurrence of disease were distributed as follows: 68 patients relapsed in the ipsilateral breast following CS + RT within 5 years of original diagnosis (EARLYBR). Fifty-one patients relapsed in the ipsilateral breast after 5 years from original diagnosis (LATEBR). Thirty-five patients relapsed in the chest wall within 5 years following mastectomy (EARLCW). Eighteen patients relapsed in the chest wall later than 5 years following mastectomy, and 41 patients failed in the regional lymphatics following mastectomy or CS + RT (REGREC). Patients with breast relapses were generally treated with salvage mastectomy, and patients with chest wall or regional nodal relapses were treated with radiation to the chest wall, regional nodes, or both. Systemic therapy at the time of local-regional relapse was highly individualized, ranging from observation to tamoxifen to high-dose chemotherapy with transplantation. With a median follow-up of 14 years, the overall 10-year survival for all 213 patients was 61%, and the 10-year distant metastasis-free rate was 59%. Patients with a LATEBR had a relatively favorable prognosis with a 5-year postrelapse distant metastasis rate of 80%. Patients with EARLYBR and LATECW had a similar prognosis, with a 5-year postrelapse distant metastasis rate of 61% and 65%, respectively. Patients with an EARLCW had a 5 year distant recurrence-free rate following a local relapse of 42%. Ten-year survivals from original diagnosis were 62% and 50%, respectively, and distant metastasis-free survival rates were 56% and 52%, respectively. Patients suffering REGREC following mastectomy or CS + RT carried a poor prognosis with a 10-year survival of 33% and a 10-year distant metastasis-free rate of 30%. Patients sustaining local-regional relapse as a first site of failure may be divided into prognostic groups. Patients with LATEBR have a relatively favorable prognosis. Patients with EARLYBR and CWREC have a poorer prognosis with a distant metastatic rate of approximately 50% within 5 years of local-regional relapse. Patients with REGREC have the poorest prognosis. Placing patients with breast cancer and local regional relapse into these prognostic categories may be helpful in decision making regarding the role of systemic therapy at the time of local-regional relapse. PMID- 11896753 TI - Age and body mass index may increase the chance of failure in sentinel lymph node biopsy for women with breast cancer. AB - Age and body mass index (BMI) have been shown to correlate with an increased incidence of failure in identifying a sentinel lymph node (SLN). Mapping senior, overweight adults is common; therefore, the relationship of patient age and BMI on SLN biopsy success is essential. This study examines the mapping failures as they relate to age and BMI. From April 1994 to May 1999, patients underwent an injection of radiocolloid (450 mci) and blue dye (5 cc) prior to SLN biopsy. SLN biopsy failure was defined as lymph nodes being unidentifiable by blue dye or having an in vivo node radiocolloid count of less than 3:1 over background count. BMI was measured as (weight in pounds)(703)/(height in inches)(2); 1,356 patients were attempted for SLN mapping, and 54 failed (3.98%). The radioactive node count was inversely proportional to age ( p < 0.0001). The radioactive node count decreased by a mean of 34 counts per node with each additional year ( p < 0.001). The estimated odds ratios for success were 0.945 for age and 0.946 for BMI. Therefore, every increase of 1 year of age or one unit of BMI decreased the odds of success by approximately 5%. The mean BMI was 29.54 in failed patients and was 26.42 in successful mapping patients ( p = 0.042). Surgeons should be aware that node counts will decrease with increasing age and that increased age and BMI are potential risk factors for SLN mapping failure. However, increased age and/or BMI alone do not appear to be contraindications for SLN biopsy in older or overweight patients. PMID- 11896754 TI - Toxicity of antiestrogens. AB - The object of this article is to review briefly the preclinical and clinical safety of some antiestrogens. Tamoxifen, toremifene, droloxifene, and idoxifene are polyphenylethylene antiestrogens, whereas the pure antiestrogen, ICI 182,780 or faslodex, as well as raloxifene, is of a different structure. Tamoxifen has been shown to be genotoxic in several studies. It induces unscheduled DNA synthesis in rat hepatocytes and micronuclei in MCL-5 a cells in vitro. Tamoxifen also induces aneuploidy in rat liver in vivo and chromosome aberrations and micronuclei in mouse bone marrow. Toremifene has also shown to be genotoxic, but to a far lower extent, by inducing micronuclei in MCL-5 a cells in vitro and by inducing aneuploidy in rat liver in vivo. Tamoxifen has been shown to be hepatocarcinogenic in the rat in at least four independent long-term studies. The initiation of tumors in the rat is the result of metabolic activation by cytochrome P450 isoenzymes to an electrophile(s) that binds irreversibly to DNA. The other antiestrogens have not been shown to be carcinogenic in rodents. In several independent clinical studies, the risk of endometrial cancer has increased among tamoxifen-treated women. After reviewing the available data, the International Agency for Research on Cancer concluded that there was sufficient evidence to show that tamoxifen is a class I human carcinogen. The increased risk for endometrial cancer occurs predominantly among women who are 50 years old or older and who have been treated with tamoxifen. It is not yet clear whether the uterine tumor formation is a result of genetic mechanisms, analogous to those seen in the rat liver or due to the estrogen agonist action of tamoxifen. However, the other antiestrogens with a more or less similar intrinsic estrogenic potential have not been shown to be carcinogenic in humans. PMID- 11896755 TI - Sentinel lymph node mapping following neoadjuvant chemotherapy for breast cancer. AB - The purpose of this study was to evaluate the feasibility of sentinel lymph node mapping in patients undergoing neoadjuvant chemotherapy for breast carcinoma prior to lumpectomy or mastectomy and sentinel lymph node mapping followed by complete axillary dissection. A retrospective analysis of 14 patients from February 1998 to July 2000 with stage I to stage IIIB breast cancer diagnosed by core biopsy underwent neoadjuvant chemotherapy (doxorubicin/cyclophosphamide) prior to definitive surgery, including lumpectomy or mastectomy and sentinel lymph node mapping, followed by full axillary dissection. Thirteen of 14 patients had successful sentinel lymph node identification (93%), and all 14 underwent full axillary dissection. An average of 2.2 sentinel nodes and a median of 16 axillary lymph nodes (including sentinel nodes) were found per patient. Of the 13 patients in whom a sentinel lymph node was identified, 10 were positive for metastases (77%). Only 4 of the 10 had further axillary metastases (40%). Three patients had negative sentinel lymph nodes shown by hematoxylin and eosin and cytokeratin stainings and had no axillary metastases (0% false negative). The single patient in whom a sentinel lymph node could not be identified had stage IIIA disease with extensive lymphatic tumor emboli. Sentinel lymph node mapping is feasible in neoadjuvant chemotherapy breast cancer patients and can spare a significant number of patients the morbidity of full axillary dissection. Further study to evaluate sentinel lymph node mapping in this patient population is warranted. PMID- 11896756 TI - Role of expression of cell cycle inhibitor p27 and MIB-1 in predicting lymph node metastasis in male breast carcinoma. AB - Tumor expression of the proliferation marker (MIB-1) and the cell cycle-related protein (p27) may predict the biologic behavior of various human tumors. The purpose of this study was to evaluate the role of p27 and MIB-1 expression in predicting lymph node metastasis in male breast carcinomas (MBCs). We studied 67 patients with invasive MBC who had undergone modified radical mastectomy. Pathologic lymph node status was correlated with the p27 protein and the MIB-1 proliferation index. These factors were studied immunohistologically by standard methods. Men in this study ranged from 36 to 92 years of age (mean, 63 years); 43 (64%) were T1 lesions, and 24 (36%) were T2 lesions. Twenty-nine patients (43%) had positive nodes. p27 was expressed in 43 tumors (64%) and MIB-1 in 13 tumors (19.4%). Tumors with positive p27 showed positive lymph nodes in 10 cases (23%). In contrast, p27-negative tumors had positive lymph nodes in 18 cases (75%). Tumors positive for MIB-1 show positive lymph nodes in 11 cases (85%). However, when MIB-1 was negative, only 16 patients (30%) had positive lymph nodes. Multivariate logistic regression analysis confirmed the utility of MIB-1 overexpression in predicting lymph node metastasis ( p < 0.0006). Also, decreased p27 protein expression strongly correlates with lymph node metastasis ( p < or = 0.0001). Furthermore, when p27 was negative and MIB-1 was positive, 100% of the patients had positive lymph nodes. In contrast, when p27 was positive and MIB-1 was negative, only 12% of patients had positive lymph nodes. The reduced expression of the p27 protein and the overexpression of the MIB-1 proliferation index in this study show a significant correlation in predicting lymph nodes metastasis in MBCs. PMID- 11896757 TI - Premenarchal athletic injury to the breast bud as the cause for asymmetry: prevention and treatment. AB - Some variation in breast size is normal and is common in most women. When this variation becomes large and appreciable asymmetry develops--greater than a one cup size difference--the asymmetry often disrupts the patient's life. The etiology of most breast asymmetries is unknown; however, current theories on causes include endocrine, iatrogenic, and traumatic injury. The Tulane University Plastic Surgery Service recently evaluated two cases of breast asymmetries that developed after traumatic injury to the breast bud while the body was under increased physical stress. Both girls sustained injuries at approximately 10 to 11 years old (Tanner Stages I-II) while participating in gymnastics. PMID- 11896759 TI - Benign phyllodes tumor of the male breast. PMID- 11896758 TI - Male ductal carcinoma in situ presenting as bloody nipple discharge: a case report and literature review. AB - Male breast carcinoma accounts for 1% of all diagnosed breast carcinoma. Pure ductal carcinoma in situ in men is extremely rare. Unfortunately, male breast cancer is often diagnosed at a late stage because of the minimal awareness of presenting symptoms by the patient and sometimes by the health care provider. Because of this late presentation, the overall prognosis is less favorable. This case is presented to emphasize the importance of recognizing bloody nipple discharge as a clinical sign of male ductal carcinoma in situ and an opportunity for early diagnosis. PMID- 11896760 TI - Breast lipoma. PMID- 11896761 TI - Surviving breast cancer. PMID- 11896762 TI - Contrast-enhanced magnetic resonance imaging has a high sensitivity for detection of breast lesions. PMID- 11896763 TI - How the gut senses its content. PMID- 11896764 TI - Interaction of cblA/adhesin-positive Burkholderia cepacia with squamous epithelium. AB - A highly transmissible strain of Burkholderia cepacia from genomovar III carries the cable pilin gene, expresses the 22 kDa adhesin (cblA +ve/Adh +ve), binds to cytokeratin 13 (CK13) and is invasive. CK13 is expressed abundantly in the airway epithelia of cystic fibrosis (CF) patients. We have now investigated whether binding of cblA +ve/Adh +ve B. cepacia to CK13 potentiates bacterial invasion and epithelial damage using bronchial epithelial cell cultures differentiated into either squamous (CK13-enriched) or mucociliary (CK13-deficient) epithelia. Three different B. cepacia isolates (cblA +ve/Adh +ve, cblA +ve/Adh -ve and cblA ve/Adh -ve) showed minimal binding to mucociliary cultures, and did not invade or cause cell damage. In contrast, the cblA +ve/Adh +ve isolate, but not others, bound to CK13-expressing cells in squamous cultures, caused cytotoxicity and stimulated IL-8 release within 2 h. By 24 h, this isolate invaded and migrated across the squamous culture, causing moderate to severe epithelial damage. A specific antiadhesin antibody, which blocked the initial binding of the cblA +ve/Adh +ve isolate to CK13, significantly inhibited all the pathologic effects. Transmission electron microscopy of squamous cultures incubated with the cblA +ve/Adh +ve isolate, revealed bacteria on the surface surrounded by filopodia by 2 h, and within the cells in membrane-bound vesicles by 24 h. Bacteria were also observed free in the cytoplasm, surrounded by intermediate filaments containing CK13. These findings suggest that binding of B. cepacia to CK13 is an important initial event and that it promotes bacterial invasion and epithelial damage. PMID- 11896765 TI - The Haemophilus ducreyi cytolethal distending toxin activates sensors of DNA damage and repair complexes in proliferating and non-proliferating cells. AB - Cytolethal distending toxins (CDTs) block proliferation of mammalian cells by activating DNA damage-induced checkpoint responses. We demonstrate that the Haemophilus ducreyi CDT (HdCDT) induces phosphorylation of the histone H2AX as early as 1 h after intoxication and re-localization of the DNA repair complex Mre11 in HeLa cells with kinetics similar to those observed upon ionizing radiation. Early phosphorylation of H2AX was dependent on a functional Ataxia Telangiectasia mutated (ATM) kinase. Microinjection of a His-tagged HdCdtB subunit, homologous to the mammalian DNase I, was sufficient to induce re localization of the Mre11 complex 1 h post treatment. However, the enzymatic potency was much lower than that exerted by bovine DNase I, which caused marked chromatin changes at 106 times lower concentrations than HdCdtB. H2AX phosphorylation and Mre11 re-localization were induced also in HdCDT-treated, non proliferating dendritic cells (DCs) in a differentiation dependent manner, and resulted in cell death. The data highlight several novel aspects of CDTs biology. We demonstrate that the toxin activates DNA damage-associated molecules in an ATM dependent manner, both in proliferating and non-proliferating cells, acting as other DNA damaging agents. Induction of apoptotic death of immature DCs by HdCDT may represent a previously unknown mechanism of immune evasion by CDT-producing microbes. PMID- 11896766 TI - Distinct protein patterns associated with Listeria monocytogenes InlA- or InlB phagosomes. AB - Internalization of Listeria monocytogenes into non-phagocytic cells is mediated by the interactions between the two bacterial invasion proteins InlA (internalin) and InlB and their cellular surface receptors E-cadherin and c-Met. To get an insight into all the cellular components necessary for uptake and early intracellular life, we undertook a global proteomic characterization of the early listerial phagosome in the human epithelial cell line LoVo. First, we proceeded to an immunocytochemical characterization of intracellular marker recruitment to phagosomes containing latex beads coated with InlA or InlB. E-cadherin and c-Met were, as expected, rapidly recruited to the phagosomal formation site. Phagosomes subsequently acquired the early endosomal antigen 1 (EEA1) and the lysosomal associated membrane protein 1 (LAMP1), while presenting a more delayed enrichment of the lysosomal hydrolase cathepsin D. Early phagosomes devoid of lysosomal, endoplasmic reticulum and Golgi enzymatic activities could then be isolated by subcellular fractionation of LoVo cells. Two-dimensional gel electrophoresis (2DPAGE) revealed differences between the protein profiles of InlA- or InlB phagosomes and those of early/late endosomes or lysosomes of naive LoVo cells. One major protein specifically recruited on the InlB-phagosomes was identified by mass spectrometry as MSF, a previously reported member of the septin family of GTPases. MSF forms filaments that co-localize with the actin cytoskeleton in resting cells and it is recruited to the entry site of InlB-coated beads. These results suggest that MSF is a putative effector of the InlB-mediated internalization of L. monocytogenes into host cells. PMID- 11896767 TI - Overexpression of the Leishmania amazonensis Ca2+-ATPase gene lmaa1 enhances virulence. AB - A gene for a Ca2+-transporting ATPase (lmaa1) from the trypanosomatid parasite Leishmania (mexicana) amazonensis was overexpressed in two clones of L. amazonensis differing in their virulence. RNA and protein expression of the gene was increased in transfectants, as was the infectivity of transfectants versus parental types in both mouse and in vitro macrophage infection experiments. The virulence of the almost avirulent clone was enhanced such that it was more virulent than the parental 'virulent' clone. Growth of the parasites in culture as promastigotes, after isolation from mouse lesions, indicated that transfection led to improved survival of promastigotes during the stationary phase of culture. As it is in this culture phase that infective metacyclic forms develop, the key role of the Lmaa1 protein may be in metacyclogenesis. The protein may be important in the synthesis and trafficking of new proteins through the secretory pathway, as we demonstrate, using a green fluorescent protein hybrid and by immunofluorescence, that the Lmaa1 protein is located in the endoplasmic reticulum in promastigotes and amastigotes of L. amazonensis. PMID- 11896768 TI - 3D in-vivo optical skin imaging for topographical quantitative assessment of non ablative laser technology. AB - BACKGROUND: A new method for treating facial rhytides and acne scars with nonablative laser and light source techniques has recently been introduced. Given the inherent limitations of photographic and clinical evaluation to assess subtle changes in rhytides and surface topography, a new noninvasive objective assessment is required to accurately assess the outcomes of these procedures. OBJECTIVE: The purpose of this study was to measure and objectively quantify facial skin using a novel, noninvasive, In-vivo method for assessing three dimensional topography. This device was used to quantify the efficacy of five treatment sessions with the 1064 nm QS Nd:YAG laser for rhytides and acne scarring, for up to six months following laser treatment. METHODS: Two subjects undergoing facial rejuvenation procedures were analyzed before and after therapy using a 30-mm, three-dimensional microtopography imaging system (PRIMOS, GFM, Teltow, Germany). The imaging system projects light on to a specific surface of the skin using a Digital Micromirror Device (DMD Texas Instruments, Irving, TX) and records the image with a CCD camera. Skin Surface microtopography is reconstructed using temporal phase shift algorithms to generate three-dimensional images. Measurements were taken at baseline, at various times during the treatment protocol, and then at three and six-month follow-up visits. Silicone skin replicas (FLEXICO, Herts, England) were also made before and after the laser treatment protocol for comparison to In-vivo acquisition. RESULTS: Skin roughness decreased by 11% from baseline after three treatment sessions in the wrinkles subject, while a 26% improvement of skin roughness was recorded by 3D In-vivo assessment six months following the fifth treatment session. The subject with acne scarring demonstrated a 33% decrease in roughness analysis after three treatment sessions by 3D In-vivo assessment. A 61% improvement in surface topography was recorded 3-months following the fifth treatment session, which was maintained at the 6-month follow-up. CONCLUSION: Three-dimensional In-vivo optical skin imaging provided a rapid and quantitative assessment of surface topography and facial fine lines following multiple treatment sessions with a 1064-nm QS Nd:YAG laser, correlating with clinical and subjective responses. This imaging technique provided objective verification and technical understanding of nonablative laser technology. Wrinkle depth and skin roughness decreased at the three and six-month follow-up evaluations by 3D In-vivo assessment, indicating ongoing dermal collagen remodeling after the laser treatment protocol. Future applications may include comparison of nonablative laser technology, optimization of treatment regimens, and objective evaluation of other aesthetic procedures performed by dermatologists. PMID- 11896769 TI - Minocycline-induced hyperpigmentation of the tongue: successful treatment with the Q-switched ruby laser. AB - BACKGROUND: Minocycline-induced hyperpigmentation (MIH) is a benign condition that may persist for years despite abrogation of therapy. The Q-switched ruby laser (QSRL) has been successful in removing such lesions from the skin. To date there is no documentation of QSRL or any laser being used to treat lingual hyperpigmentation associated with minocycline therapy. OBJECTIVE: Long-term follow-up results are reported for the use of QSRL to treat lingual hyperpigmentation. The literature is reviewed comparing the use of different laser systems on MIH. METHODS: A 26-year-old woman with pigment changes of the tongue and buccal mucosa due to long-term minocycline therapy was treated with four consecutive sessions with QSRL (694 nm, 20-nsec pulse duration, and 6.5 mm spot size) at 3.6-4.0 J/cm2. RESULTS: A 90% resolution was achieved after three treatments. After the final treatment the lesions were completely gone. There were no side effects reported. No new pigment was detected at follow-up. CONCLUSION: Treatment with the QSRL is a safe and effective strategy for treating hyperpigmentation of the tongue associated with minocycline therapy. PMID- 11896770 TI - Dilute povidone-iodine solutions inhibit human skin fibroblast growth. AB - BACKGROUND: Povidone-iodine solutions are widely used and highly effective antiseptics. Although commonly used at full strength, this concentration appears to be toxic to the cells involved in wound healing. Few systematic studies of povidone-iodine toxicity have been reported. The effects of various dilutions of 10% povidone-iodine solution on the growth of human diploid fibroblasts were assessed using in vitro cell culture. OBJECTIVE: The purpose of this study was to systematically evaluate the toxicity of povidone-iodine on living cells using an in vitro model. METHODS: Adult skin fibroblasts and fetal lung fibroblasts were subcultivated at various seeding densities of 3000-10,000 cells/cm2 and grown in polystyrene tissue culture flasks under an atmosphere containing 5% O2, 5% CO2, and 90% N2. Cells were grown in a medium containing various concentrations of povidone-iodine (1%, 0.1%, 0.025%, 0.01%, and 0%). Cell attachment was reduced by 0.1% and 1% povidone-iodine in our initial studies; subsequent experiments were performed by changing the medium to contain the povidone-iodine 24 hours after seeding. Growth curves were performed by counting triplicate cultures every 48 hours for 250-300 hours. RESULTS: Fibroblast growth was progressively retarded at 0.01% and 0.025%, and totally inhibited by 0.1% and 1% povidone-iodine solutions. Partial recovery of cell growth after limited exposure of cultures to dilute solutions of povidone-iodine was noted. CONCLUSION: This study shows that even dilute solutions of povidone-iodine are toxic to human fibroblasts. The results indicate that caution should be used when povidone-iodine is placed on an open wound, and that prolonged contact with viable uncontaminated tissue should be avoided. PMID- 11896771 TI - Adult-onset facial colloid milium successfully treated with the long-pulsed Er:YAG laser. AB - BACKGROUND: Adult colloid milium is a rare cutaneous deposition disorder that frequently involves areas of chronic sun exposure, especially the face and dorsal hands. Attempts to remove these lesions are generally unsuccessful, but dermabrasion has been reported to be effective. OBJECTIVE: To present an effective therapeutic alternative to dermabrasion for facial colloid milium. METHODS: A 41-year-old man with extensive facial colloid milium underwent full face resurfacing with a long-pulsed Er:YAG laser (9.8 J/cm2, 5 mm spot, 10-msec pulse duration). Additional passes were performed over the areas of dense colloid milium to achieve approximately 80% ablation of lesions.results. At the 7-month follow-up there was no scarring, textural changes, dyspigmentation, or clinical evidence of recurrence. CONCLUSION: Long-pulsed Er:YAG laser should be considered an effective alternative to dermabrasion for facial colloid milium. PMID- 11896772 TI - Nd:YAG laser (1064 nm) irradiation for lower extremity telangiectases and small reticular veins: efficacy as measured by vessel color and size. AB - BACKGROUND: Laser treatment of lower extremity telangiectases and small reticular veins has remained difficult because of vessel color, diameter, depth, and associated high-pressure flow. Traditionally, larger-caliber blue leg veins do not respond well to laser treatment. Nd:YAG laser (1064 nm) irradiation is absorbed by oxyhemoglobin and reduced hemoglobin and is associated with greater depth of penetration than other previously studied vascular lasers. OBJECTIVE: To evaluate a millisecond contact-cooled 1064 nm Nd:YAG laser for the treatment of telangiectases and small reticular veins. METHODS: Twenty-one lower extremity sites, with Fitzpatrick skin types I-IV, received two laser treatments separated by a 4 to 6-week period. Blue and red vessels, ranging in size from 0.25 to 4.0 mm were treated. Pulse durations of 10-50 msec were utilized at fluences of 90 187 J/cm2. Three months after the last treatment, patients were evaluated for vessel improvement and complications. RESULTS: Seventy-one percent of lower extremity vessels had improvement graded as significant. All vessel colors and sizes were successfully treated. The only complication at 3 months was postinflammatory hyperpigmentation. CONCLUSION: 1064 nm Nd:YAG laser irradiation with associated contact cooling is a safe and effective treatment for telangiectases and small reticular veins of the lower extremities. PMID- 11896773 TI - A side-by-side comparative study of 1064 nm Nd:YAG, 810 nm diode and 755 nm alexandrite lasers for treatment of 0.3-3 mm leg veins. AB - BACKGROUND: Laser and intense pulsed light device treatments of leg veins have generally yielded disappointing results. Use of longer wavelengths, longer pulse widths, and better cooling devices have recently sparked renewed interests in these methods. OBJECTIVE: To prospectively compare, side by side, a 3-msec cryogen spray-equipped 755 nm alexandrite, a sapphire window cooled super-long pulse 810 nm diode, and a variable pulse width, cryogen spray-equipped 1064 nm Nd:YAG laser for the treatment of 0.3-3mm leg veins. METHODS: Thirty female volunteers, skin types I-V, age 32-67 years with comparable sets of leg veins were treated with the Nd:YAG laser and either the diode laser, alexandrite laser, or both. In most patients two to three sets of comparable sites were treated. Treatment parameters varied with each laser and according to the size of veins being treated. Patients were examined 1 week after each treatment and at 1, 2, and 3 months after the last treatment. Pre- and posttreatment 35mm photographs were taken. Improvement was judged by two experienced physicians both visually on patients and by comparison of pre- and posttreatment photographs. Results were graded as percent resolution, in five groups, 0%, 0-25%, 25-50%, 50-75%, and 75 100%. RESULTS: In the 22 patients completing the study, 36 leg vein sites were treated with the Nd:YAG laser, 18 leg vein sites were treated with the diode laser, and 12 leg vein sites were treated with the alexandrite laser. Greater than 75% improvement was observed at 88% of the Nd:YAG laser-treated sites, 29% of the diode laser-treated sites, and 33% of the alexandrite laser-treated sites. Greater than 50% improvement was observed at 94% of the Nd:YAG laser-treated sites, 33% of the diode laser-treated sites, and 58% of the alexandrite laser treated sites. Less than 25% improvement was observed at 6% of the Nd:YAG laser treated sites, 39% of the diode laser-treated sites, and 33% of the alexandrite laser-treated sites. Pain during treatment was variably perceived by patients, but occasionally sufficient for patients to decline further treatment. Posttreatment purpura and telangiectatic matting were a significant drawback for the alexandrite laser. Transient hemosiderin pigmentation, as seen with sclerotherapy, was common with larger vessels. CONCLUSION: The cryogen spray equipped 1064 nm Nd:YAG laser was remarkably effective and safe for the treatment of 0.3-3 mm leg veins. The use of topical anesthesia may be needed for some patients. The super-long-pulse 810 nm diode laser gave unpredictable results. Additional refinements of fluence and pulse width could improve its performance. The 3-msec, 755 nm alexandrite laser at fluences of 60-70 J/cm2 and an 8 mm spot can be effective, but inflammatory response, purpura, and matting limit its usefulness. Longer pulse widths might decrease these problems. For leg vein treatment, the 1064 nm wavelength is very safe for type V skin, the 810 nm wavelength at super-long pulse widths of 400-1000 msec is very safe for type IV and marginal for type V skin, and the 755 nm wavelength is limited to nontanned type I-III skin. PMID- 11896774 TI - Double-blind, half-face study comparing topical vitamin C and vehicle for rejuvenation of photodamage. AB - BACKGROUND: Aging of the population, in particular the "baby boomers," has resulted in increased interest in methods of reversal of photodamage. Non invasive treatments are in high demand, and our knowledge of mechanisms of photodamage to skin, protection of the skin, and repair of photodamage are becoming more sophisticated and complex. OBJECTIVE: The objective of this study is to determine if the topical use of a vitamin C preparation can stimulate the skin to repair photodamage and result in clinically visible differences, as well as microscopically visible improvement. METHODS: Ten patients applied in a double blind manner a newly formulated vitamin C complex having 10% ascorbic acid (water soluble) and 7% tetrahexyldecyl ascorbate (lipid soluble) in an anhydrous polysilicone gel base to one-half of the face and the inactive polysilicone gel base to the opposite side. Clincial evaluation of wrinkling, pigmentation, inflammation, and hydration was performed prior to the study and at weeks 4, 8, and 12. Two mm punch biopsies of the lateral cheeks were performed at 12 weeks in four patients and stained with hematoxylin and eosin, as well as in situ hybridization studies using an anti-sense probe for mRNA for type I collagen. A questionnaire was also completed by each patient. RESULTS: A statistically significant improvement of the vitamin C-treated side was seen in the decreased photoaging scores of the cheeks (P = 0.006) and the peri-oral area (P = 0.01). The peri-orbital area improved bilaterally, probably indicating improved hydration. The overall facial improvement of the vitamin C side was statistically significant (P = 0.01). Biopsies showed increased Grenz zone collagen, as well as increased staining for mRNA for type I collagen. No patients were found to have any evidence of inflammation. Hydration was improved bilaterally. Four patients felt that the vitamin C-treated side improved unilaterally. No patient felt the placebo side showed unilateral improvement. CONCLUSION: This formulation of vitamin C results in clinically visible and statistically significant improvement in wrinkling when used topically for 12 weeks. This clinical improvement correlates with biopsy evidence of new collagen formation. PMID- 11896775 TI - A composite nasolabial flap for an entire ala reconstruction. AB - BACKGROUND: It is difficult to reconstruct an entire alar defect. We report a case of an entire alar reconstruction carried out in two stages with a composite nasolabial flap lined with retroauricular skin and supported with the attached conchal cartilage. OBJECTIVE: A composite nasolabial flap was applied for reconstruction of entire alar defect in this patient. METHODS: The lining skin for the flap was obtained from retroauricular region acceptable to the patient, and the conchal cartilage was used simultaneously for support and as the attachment for the lining skin. The flap was then replaced and secured. Half a year later, the flap was flipped and transferred to the alar defect as a second step. RESULTS: The final shape and texture were satisfactory. All procedures were performed under local anesthesia. CONCLUSION: Our design of composite flap has been successfully utilized to repair an entire alar defect with cosmetically and functionally good results, minimizing the donor area and the resulting operation scars. This composite nasolabial flap is thought to be the best choice of a flap for an entire alar defect reconstruction. PMID- 11896776 TI - Localized melanosis in an immunocompromised patient with local metastatic melanoma. AB - BACKGROUND: Melanosis is a rare, but well recognized late complication of some cases of metastatic melanoma. The precise pathogenesis is unclear. Most described cases report diffuse metastases and diffuse melanosis. OBJECTIVE: To present a case of localized melanosis secondary to locally metastatic melanoma without evidence of systemic involvement in a renal transplant patient. METHODS: Case report and literature review. RESULTS: We describe a case of localized melanosis in an immunocompromised patient without evidence of systemic metastases. The patient underwent limb perfusion and her immunosuppressive therapy has been stopped. The melanosis has not regressed; however, there has been no systemic involvement and her transplanted kidney is still functioning well. CONCLUSION: To our knowledge this is the first report of this phenomenon in an immunocompromised host. PMID- 11896777 TI - Treatment of face veins with a cryogen spray variable pulse width 1064 nm Nd:YAG Laser: a prospective study of 17 patients. AB - BACKGROUND: Through the 1990s laser treatment of leg veins has been a challenge. Newer, longer wavelength lasers with variable pulse width have shown promising results for both telangiectasia and reticular leg veins. Experience with these lasers for treatment of facial telangiectasia and periorbital reticular veins is scant. OBJECTIVE: To our knowledge this is the first prospective study to evaluate a variable pulse width, cryogen spray-equipped 1064 nm Nd:YAG laser for the treatment of facial telangiectasia and larger periorbital reticular veins. METHODS: Seventeen volunteers, ages 32-67 years, skin types I-IV, with facial telangiectasia and reticular periorbital/temporal veins were treated once with the Nd:YAG laser at fluences of 125-150 J/cm2 with a 6 mm spot size and pulse widths of 25 msec for small vessels and 75-100 msec for reticular veins. Patients were examined 1 week and 1 month after the treatment. Pre- and posttreatment 35mm photographs were taken. Improvement was judged by two experienced physicians, both visually on patients and by comparison of pre- and posttreatment photographs. Results were graded as percent resolution, in five groups: 0%, 0 25%, 25-50%, 50-75%, and 75-100%. RESULTS: All 17 patients completed the study. Thirty-two sites were treated (24 cheek, nose, and chin telangiectases, and 8 periorbital reticular veins). Greater than 75% improvement was observed in 97% of the treated sites. Greater than 50% improvement was observed in 100% of the treated sites. Notably, 100% of the facial reticular veins treated have essentially 100% resolution. Pain during treatment was variably perceived by patients, but occasionally sufficient for patients to express reservations about additional future treatment. Transient erythema and edema were common, but fine crusting was rare. Small areas of purpura were also quite rare. CONCLUSION: The cryogen spray-equipped 1064 nm Nd:YAG laser was remarkably effective and safe for skin type I-IV patients. It is an excellent tool for treatment of facial telangiectasia with minimal risk of purpura. For the first time we appear to have a simple, safe, and effective treatment for 1-2 mm periorbital reticular veins. The use of topical anesthesia may be needed for some patients. PMID- 11896778 TI - Dermatologic surgery and the pregnant patient. AB - BACKGROUND: Because of concerns about potential harm to the mother or fetus, dermatologic surgeons are frequently hesitant to perform cutaneous surgery on pregnant patients. OBJECTIVE: To review the relevant physiologic changes during pregnancy, appropriate preparation for and timing of procedures, and drug safety. METHODS: A literature review was performed of dermatologic and nondermatologic journals discussing physiology, surgery, and drug safety in the pregnant patient. RESULTS: Special positioning is required for the pregnant patient during surgery. Low doses of most local anesthetics with epinephrine as well as nitrous oxide less than 50% are safe to use during pregnancy. Sedatives and opioids are potential teratogens and should be avoided. Safe antibiotics to use during skin surgery in pregnancy include penicillins, cephalosporins, and nonestolate erythromycin. If necessary, lymph node dissections under general anesthesia in the pregnant melanoma patient should occur during the second trimester. CONCLUSION: With appropriate preparation, safe and successful cutaneous surgery can be performed on the pregnant patient. PMID- 11896779 TI - The corset platysma repair: a technique revisited. AB - BACKGROUND: Platysma banding along with excess submental adipose tissue and sagging skin can lead to an aged appearance. Several methods for improving neck and submental contours exist, including neck liposuction, bilateral platysma plication, midline platysma plication with transection of distal fibers, necklift with skin excision, and botulinum toxin injection for platysma relaxation. With the current interest in minimally invasive procedures, surgeons and patients are searching for techniques that produce maximal improvement with minimal intervention. OBJECTIVE: To present a modified technique for maximizing neck contouring, discuss possible complications of the procedure, and describe appropriate candidates for the corset platysmaplasty. METHODS: We performed a retrospective analysis of 10 consecutive patients who underwent neck liposuction with concomitant corset platysmaplasty at our institution. RESULTS: All 10 patients achieved good to excellent submental and jawline contouring, determined by both physician and patient assessment, with no visible platysma banding at 6 months follow-up. No major complications were noted. CONCLUSION: Use of corset platysmaplasty is a safe and effective method for neck rejuvenation. This variation of platysmaplasty can be used in conjunction with neck liposuction to maximize jawline and neck contour enhancement. PMID- 11896780 TI - A liquid adhesive bandage for the treatment of minor cuts and abrasions. AB - BACKGROUND: Octyl-2-cyanoacrylate is U.S. Food and Drug Administration (FDA) approved for the closure of incisions and lacerations. In animal studies, a more flexible formulation of octyl-2-cyanoacrylate suitable for cuts and abrasions produced faster healing of partial thickness wounds than traditional bandages. OBJECTIVE: To evaluate the effectiveness of a more flexible octyl-2-cyanoacrylate liquid adhesive bandage for the treatment of minor cuts and abrasions. METHODS: One hundred sixty-two volunteers with recent minor cuts or abrasions were recruited and randomized to treatment with either liquid adhesive bandage (LAB) or a control device (Band-Aid brand adhesive bandage, sheer, 2.5 cm). The primary efficacy criterion was complete healing at day 12. Secondary efficacy criteria were the ability of patients to properly apply LAB, and the ability of LAB to stop bleeding, to reduce pain, and to remain on the wound. RESULTS: At day 12 there was no statistical difference between the number of completely healed wounds in the LAB and the bandage-treated patients (P =.493). The ability of patients, as rated by investigators, to effectively apply the LAB device and the bandage was not significantly different (P =.165). Only the LAB provided significant hemostasis (P =.0001) and pain relief (P =.002). CONCLUSION: In this randomized, controlled trial, the LAB was as effective as the control at promoting healing as measured by complete healing at day 12. The LAB was easy to use and gave rapid control of bleeding and pain, forming a film that stayed on wounds well. PMID- 11896781 TI - Prognostic factors for metastasis in squamous cell carcinoma of the skin. AB - BACKGROUND: Squamous cell carcinoma (SCC) of the skin exhibits a significant propensity to metastasize. A number of variables have been reported to influence the tendency of SCC to metastasize. Because of the increasing incidence of skin cancer, it is becoming increasingly important to identify those neoplasms which are biologically more aggressive. We report 25 cases of metastatic SCC and compare them to 175 cases of nonmetastasizing SCC treated during the same period. OBJECTIVE: To characterize tumors with the greatest tendency to metastasize. METHODS: A tumor registry from the Dermatologic Surgery Unit at the Medical University of South Carolina was accessed to obtain records on 200 patients diagnosed with invasive SCC managed by Mohs surgery from 1988 to 1998. A retrospective analysis was conducted. The characteristics of patients with metastatic SCC and those with nonmetastatic SCC were compared using the chi squared test and Fisher's exact test. RESULTS: Of 200 tumors, 25 (12.5%) metastasized. Size, Clark's level, degree of differentiation, the presence of small tumor nests, infiltrative tumor strands, single-cell infiltration, perineural invasion, acantholysis, and recurrence all correlated strongly with metastasis. Location, ulceration, inflammation, and Breslow depth did not correlate with the development of metastasis. CONCLUSION: Patients with tumors that exhibit certain clinical and histologic features are more likely to metastasize and need close follow-up to detect recurrence and metastasis early, allowing for appropriate life-saving intervention. Sentinel lymph node biopsy should be considered in patients with high-risk SCC. PMID- 11896782 TI - Factors that influence healing in chronic venous ulcers treated with cryopreserved human epidermal cultures. AB - BACKGROUND: Cryopreserved epidermal cultures (CEC) offer an "off the shelf" treatment for chronic wounds. These cultures are derived from neonatal foreskin and grow rapidly in vitro to form epidermal sheets. They do not require a biopsy from the patient, an advantage compared to autografts. They seem to act as a biological dressing, stimulating epithelialization from the wound edges and adnexae, probably through growth factor release. OBJECTIVE: To summarize our recent experience with the use of CEC in chronic venous leg ulcers and to determine the factors that influence healing in chronic venous ulcers treated with CEC. PATIENTS AND METHODS: A single arm, open label study including a total of 11 patients with documented venous ulcers was performed. The study involved the application of cryopreserved epidermal cultures every other week to nonhealing leg ulcers for a total of 12 weeks or until complete healing of the ulcer. RESULTS: A total of 11 patients with one or more leg ulcers were treated. The average age was similar in healed and unhealed groups. Seven patients completely healed after an average of 4.14 CEC applications. Four patients did not heal after a total of 12 CEC applications. CONCLUSION: Predictors for failure to heal after CEC application in our patients were long wound duration, wound size, presence of lipodermatosclerosis, and history of failed prior split thickness skin grafts. PMID- 11896783 TI - Interview with Dr. Hugo Partsch (by Jose Antonio Olivencia). AB - Is it necessary to write an introduction for a man whose name is familiar to every phlebologist and angiologist, and who is currently the President of the International Union of Phlebology? (Figure 1) The list of Professor Hugo Partsch's publications (more than 360) and his memberships in scientific committees and advisory boards would fill several pages. In the early 1970s I was reading, with great interest, his articles in "VASA," and several of them have become part of my basic medical knowledge. In all these years, the flow of numerous publications has not diminished, now covering the entire spectrum of angiology. Just by looking to his enormous scientific production, the average reader would probably picture the author as a "typical textbook scientist" caught in a strange world consisting of vessels, either too wide or too narrow. Anyone who has met Hugo Partsch will agree that the contrary is true. It must be assumed that most readers have already seen him as a regular guest speaker at international meetings. It is still a mystery to me how he finds time for all his scientific work, at the same time overseeing the University Clinic in Vienna and traveling from one Congress to the next. Readers will know him as a highly respected scientist, as a fair and diplomatic discussant that speaks several languages, and last but not least, an untiring skier and dancer. Hugo Partsch has helped to bring European Angiology to the United States, and using his broad knowledge and Viennese charm, to merge the diverse studies and interests of phlebology, angiology, lymphology and dermatology.He was born in Vienna in 1938 where he completed all of his studies. He graduated in 1962 and served three years as a resident in southern Austria. In 1965 he continued his residency at the Wilhelmine Spital; at this hospital he would become the head of the dermatological department in 1987. Starting his training in dermatology in 1966, he became a senior registrar in 1970. He acquired the Venia Legendi in 1978 and received the title of Professor of Dermatology at the University of Vienna in 1985. He is the father of three children and has four grandchildren. One could simply define Hugo Partsch as Professor of Dermatology in Vienna. But he is the only dermatologist, to my knowledge, having done so much in the fields of thrombosis, lymphology, peripheral neuropathies, phlebology and angiology in general. PMID- 11896785 TI - Ultrapulse CO2 used for the successful treatment of basal cell carcinomas found in patients with basal cell nevus syndrome. AB - BACKGROUND: Basal cell nevus syndrome (BCNS) is an inherited condition marked by multiple basal cell carcinomas (BCCs) associated with several other abnormalities. Various treatment modalities have been used to eradicate these tumors. However, recurrences and scarring limit their use. OBJECTIVE: To evaluate the treatment of multiple BCCs associated with BCNS. METHODS: We describe three cases of BCNS in which multiple BCCs were effectively treated with ultrapulse CO2 laser. Postoperative Mohs micrographic surgical sections (thin sections looking for residual tumor) verified complete histologic clearance of the tumors. RESULTS: All three patients were effectively treated with ultrapulse CO2 laser. Minimal scarring was noted at follow-up. CONCLUSION: Ultrapulse CO2 laser can be used to effectively treat small BCCs in low-risk areas associated with BCNS with minimal posttreatment scarring. PMID- 11896784 TI - Trichilemmal carcinoma in an African American. AB - BACKGROUND: Trichilemmal carcinoma is an uncommon cutaneous neoplasm found primarily on the sun-exposed skin of the elderly. OBJECTIVE: To report a case of trichilemmal carcinoma presenting as a forehead nodule in an African American patient. METHODS: Case report and review of the literature. RESULTS: Our patient is the first reported case in the English language literature of trichilemmal carcinoma in an African American individual with skin phototype VI. CONCLUSION: The occurrence of trichilemmal carcinoma in our African American patient suggests the diagnosis of trichilemmal carcinoma be added to the differential diagnosis of a nodule on the head region in this patient population. PMID- 11896786 TI - Milia en plaque: a case report and review of the literature. AB - BACKGROUND: Milia en plaque (MEP) is an unusual entity with a distinctive clinicohistologic appearance. Optimal treatment is unestablished, particularly for MEP located on difficult anatomic areas. OBJECTIVE: To illustrate by a case report the clinical presentation and management of MEP. METHODS: A 35-year-old woman with numerous tiny cysts within an slightly erythematous base in unilateral periorbital distribution is described. Histology revealed epidermal cysts arranged within the entire dermis. RESULTS: Treatment with chemical exfoliating agents, manual extraction, and topical photodynamic therapy resulted in partial improvement. CONCLUSION: Dermatologists should become familiar with this peculiar condition, being significantly rare possibly because of underreporting or misdiagnosis. New treatment procedures must be tried to achieve a successful cosmetic result with minimal risks. PMID- 11896787 TI - Hypertrophic scars after therapy with CO2 laser for treatment of multiple cutaneous neurofibromas. AB - BACKGROUND: CO2 laser surgery is a treatment modality for cutaneous neurofibromas. OBJECTIVE: Hypertrophic and atrophic scars can result from treatment with CO2 laser surgery. We present a case of cutaneous neurofibromatosis that developed hypertrophic scars postoperatively. METHODS: Continuous wave CO2 laser surgery therapy was applied to the patient. RESULTS: Hypertrophic scars developed 2 months after therapy. CONCLUSION: With a preliminary test treatment the patient is able to see the expected result. PMID- 11896788 TI - Lessons on dermoscopy. PMID- 11896804 TI - The relationship between levels of mood, interest and pleasure and 'challenging behaviour' in adults with severe and profound intellectual disability. AB - BACKGROUND: Research on affective disorders in adults with intellectual disability (ID) suggests that depression may not present a 'classic picture' in individuals with severe and profound ID, but may include challenging behaviours, which are referred to as 'atypical symptoms', such as self-injury, aggression and irritability. The aim of the present study was to explore whether there is an association between constructs relating closely to the core symptoms of depression and challenging behaviours in adults with severe and profound ID. METHOD: Mood and levels of interest and pleasure were measured in 53 adults with severe or pro-found ID using the Mood, Interest and Pleasure Questionnaire (MIPQ). RESULTS: Two groups of adults were identified based on MIPQ scores: (1) a 'low mood' group (lowest score = 12); and (2) a comparison group (highest scoring = 12). The groups were clearly differentiated on the MIPQ (P < 0.0001), but were comparable on age, gender and medication use. The Challenging Behaviour Interview showed no difference between the two groups in self-injury, aggression or disrupting the environment. A secondary analysis revealed that participants who showed challenging behaviour scored significantly lower on the MIPQ than those who did not show challenging behaviour. CONCLUSIONS: Possible reasons for these results and considerations for future studies are discussed. PMID- 11896805 TI - Changes in explicit memory associated with early dementia in adults with Down's syndrome. AB - BACKGROUND: A modified version of the Selective Reminding Test (SRT) (Buschke 1973) was used to examine the changes in memory that occur with early-stage dementia of the Alzheimer's type (DAT) in adults with intellectual disability (ID) and Down's syndrome (DS), and to compare these changes to those occurring with 'normal' ageing. METHOD: Hierarchical linear modelling analyses showed steep declines in the performance of participants who had met the criteria for the onset of DAT. Non-demented participants also showed declines in performance which were related to their age. However, the absolute magnitude of these declines was consistent with a 'normal' ageing pattern and not with undetected dementia. RESULTS: In analysing the specific memory components that are compromised, the present authors found that participants with early-stage DAT showed severely diminished long-term storage and retrieval processing abilities compared to their non-demented peers. Notably, these declines preceded other symptoms of dementia, in most cases by more than a full year and sometimes by as much as 3 years. CONCLUSIONS: Thus, the present results clearly confirm that memory processes are affected during early dementia in adults with DS, and that the SRT has promise as a clinical tool. PMID- 11896806 TI - Feasibility, reliability and validity of the Spanish version of Psychiatric Assessment Schedule for Adults with Developmental Disability: a structured psychiatric interview for intellectual disability. AB - BACKGROUND: Over 30% of people with intellectual disability (ID) have a comorbid psychiatric disorder. However, there are few assessment instruments available for international use and cross-cultural validation studies of these instruments are rare. The aim of the present study was to standardize the Spanish version of the Psychiatric Assessment Schedule for Adults with Developmental Disability (PAS-ADD 10), a semi-structured interview for people with ID. METHODS: After a conceptual translation, feasibility (i.e. applicability, acceptability and practicality) and reliability analyses were carried out. The predictive validity of the PAS-ADD-10 CATEGO-5 codings was also examined (i.e. positive and negative predictive values). Four independent raters with wide-ranging experience in quantitative evaluation and psychiatric assessment of ID evaluated a sample of 80 subjects with ID and borderline intellectual functioning at the AFANAS occupational centre in Jerez, Southern Spain. The ICD-10 codes were used for psychiatric diagnosis. RESULTS: The practicality of the PAS-ADD-10 is limited because of the need for previous standardization of SCAN interviews. Nevertheless, its overall feasibility was judged adequate by raters and the PAS-ADD-10 was considered extremely useful for training. Test-retest and inter-rater reliability kappa values were moderate to high. The CATEGO coding showed limited validity because of overdiagnosis of anxiety disorders and underdiagnosis of mood and psychotic disorders (positive predictive value = 74%, negative predictive value = 76%). CONCLUSIONS: The PAS-ADD-10 is a useful tool for standard psychiatric assessment of people with ID; however, CATEGO codings show low validity and a series of modifications should be considered before this instrument is used extensively in Spain. In this regard, a study on the clinical usefulness of the PAS-ADD-10 in patients with ID and severe mental disorders has been undertaken. PMID- 11896807 TI - Psychometric evaluation of a Swedish version of the Reiss Screen for Maladaptive Behavior. AB - BACKGROUND: People with intellectual disability (ID) are afflicted by mental health problems to a greater extent than other individuals, and the coexistence of ID and mental health problems involves diagnostic as well as treatment difficulties. METHODS: A Swedish version of the Reiss Screen for Maladaptive Behavior (RSMB), an instrument used for identification of mental health problems in people with intellectual disability (ID) was evaluated in terms of inter-rater agreement, internal consistency, item grouping and criterion validity based on a random sample and a clinical group of adults with ID. RESULTS: The Swedish version of the RSMB had moderate-to-low inter-rater agreement on specific items and good internal consistency. The total score was considered to be a fairly reliable measure of a positive or negative result on the RSMB. A principal component analysis yielded seven interpretable components. A close resemblance in sets of items between the Swedish version and the original version of RSMB was found for three subscales: Aggressive Behaviour, Avoidance Disorder and Depression (Behavioural Signs). The outcome of the criterion validity analysis indicated a higher rate of false negatives than false positives. CONCLUSIONS: The potential influence of concurrent psychopharmacological treatment is discussed. It is concluded that the Swedish version of the RSMB can be used as intended by staff as a primary screening device for the identification of mental health problems among people with ID in a Swedish setting. PMID- 11896808 TI - Intellectual disability amongst people on probation: prevalence and outcome. AB - BACKGROUND: Previous research has suggested that people with intellectual disability (ID) can be found in the criminal justice system. However, little is currently known about those supervised in the community on probation orders. METHODS: Ninety people on probation in south-east England were screened using the Learning Disabilities in the Probation Service (LIPS) screening tool, which was designed to be used by probation officers to identify those with possible ID. The LIPS incorporates two measures of cognitive functioning: (1) the Quick Test (QT); and (2) the Clock Drawing Test (CDT). RESULTS: Seven per cent of participants were found to have QT IQs and CDT scores 1.6 SD below the mean. When compared to those who appeared to be functioning within the normal range, those with possible ID were found to be significantly younger. However, no differences were found between the two groups in terms of the final outcome of the probation order. CONCLUSIONS: It seems likely that the probation service contains a significant minority of people with ID. Despite the fact that no difference was found to exist in terms of outcome, people with ID or borderline ID are likely to have a number of support needs which could affect the success of their time on probation. PMID- 11896809 TI - Behaviour/mental health problems in young adults with intellectual disability: the impact on families. AB - BACKGROUND: The present authors studied the impact of dual diagnosis [i.e. intellectual disability (ID) and mental disorder] in young adults on their mothers' perceived levels of stress and decisions about placement. METHODS: The mothers of 103 young adults with severe ID were interviewed using a 2-3-h in depth protocol of measures designed to assess their child's adaptive functioning, maladaptive behaviour, mental health problems and negative impact on the family, as well as their own thoughts on out-of-home placement. The Scales of Independent Behavior--Revised Problem Behavior Scale assessed problem behaviours and the Reiss Screen assessed mental disorder. RESULTS: These measures were highly correlated (r = 0.64), but tapped some different domains of maladaptive behaviour and proved to be most predictive when employed together. Behaviour and/or mental health (B/MH) problems significantly predicted the mothers' perceived negative impact of the young adult on the family, even after controlling for other young adult characteristics. These problems also predicted the family's steps toward seeking out-of-home placement, as did better young adult health and the mother's higher educational attainment; stress did not predict additional variance in placement once these variables were accounted for. CONCLUSIONS: The discussion focuses on the implications for service provision to families of young adults with B/MH problems. PMID- 11896810 TI - Mental health services and young people with intellectual disability: is it time to do better? AB - BACKGROUND: There is evidence that the mental health needs of children with disabilities are inadequate. The aim of the present study was to determine the extent of specialist health service use during adolescence by a group of individuals with intellectual disability (ID) and mental health problems. METHOD: The study population consisted of 80 young people with ID, who were examined in childhood and adolescence for psychiatric and behaviour disorder. These young people were interviewed again in early adult life for the presence of psychiatric and behaviour disorder. Evaluation questionnaires were used during the follow-up study to assess service use from adolescence. RESULTS: The key finding was that the great majority (64%) of subjects with persistent challenging behaviour from childhood into adult life and those with an established childhood psychiatric disorder received no specialist mental health care. CONCLUSIONS: The development of mental health services for this vulnerable group with complex psychiatric and behaviour disorders has been poor for a number of reasons, including lack of recognition at the primary care level and insufficient numbers of trained professionals within specialist services. PMID- 11896811 TI - Mental health of primary family caregivers with children with intellectual disability who receive a home care programme. AB - BACKGROUND: The aims of the present study were to describe the change in mental health over time in a group of family caregivers with a child with intellectual disability (ID) and to explore the effect of a home care service on the psychological well-being of the caregiver. METHODS: The authors identified children with ID who received home care services in the southern part of Taiwan. A total of 46 primary family caregivers (age range = 21-65 years) were recruited for the present study. The study design was a quasi-experimental follow-up analysis. The children with ID and their families regularly received home-based care. The 12-item version of the Chinese Health Questionnaire (CHQ) was used to evaluate the subjects' mental health at three time points: (1) baseline, (2) 3 months and (3) 9 months. The validity and reliability of the CHQ have been tested in Taiwan. The Generalized Estimating Equation was used to conduct longitudinal data analyses. RESULTS: The authors found that the family caregivers showed a significant improvement in their mental health by month 9. CONCLUSIONS: The preliminary findings of this study accredit the effect of home care services and suggest that home care services are necessary for family caregivers. PMID- 11896812 TI - People with intellectual disability, sensory impairments and behaviour disorder: a case series. AB - BACKGROUND: Sensory impairments (SIs) are more prevalent in people with intellectual disability (ID). Both conditions lead to higher rates of emotional and behavioural problems than in the general population. The identification of psychiatric disorders in this group can be difficult, particularly in those with severe ID and limited communication skills. METHOD: The present paper presents a series of 18 case reports of individuals with ID, SI and behavioural problems. RESULTS: The majority of cases were young male caucasians with congenital rubella syndrome and autistic spectrum disorder, referred because of self-injurious behaviour (SIB) or aggression. Nine cases were treated with autidepressants, five underwent environmental changes and two had medication reduced. All showed some improvement. CONCLUSIONS: The benefits of comprehensive assessments, the use of standardized assessment tools and trials of treatments are discussed in the context of making psychiatric diagnoses. PMID- 11896813 TI - Behavioural aspects of Pollitt syndrome: a 32-year follow-up of a case described by R. J. Pollitt and colleagues in 1968. AB - In 1968, R. J. Pollitt and colleagues described a syndrome characterized by abnormally brittle, sulphur-deficient hair (trichothiodystrophy), intellectual disability (ID) and growth retardation. One of the two siblings originally described by the above authors has recently been re-assessed by the present authors following a referral for advice about ritualistic behaviours. Her current clinical features are described, and the literature concerning trichothiodystrophy and ID is summarized. PMID- 11896814 TI - The Edinburgh Principles with accompanying guidelines and recommendations. AB - A panel of experts attending a 3-day meeting held in Edinburgh, UK, in February 2001 was charged with producing a set of principles outlining the rights and needs of people with intellectual disability (ID)and dementia, and defining service practices which would enhance the supports available to them. The Edinburgh Principles, seven statements identifying a foundation for the design and support of services to people with ID affected by dementia, and their carers, were the outcome of this meeting. The accompanying guidelines and recommendations document provides an elaboration of the key points associated with the Principles and is structured toward a four-point approach: (1) adopting a workable philosophy of care; (2) adapting practices at the point of service delivery; (3) working out the coordination of diverse systems; and (4) promoting relevant research. It is expected that the Principles will be adopted by service organizations world-wide, and that the accompanying document will provide a useful and detailed baseline from which further discussions, research efforts and practice development can progress. PMID- 11896815 TI - Oral manifestations in 101 Cambodians with HIV and AIDS. AB - BACKGROUND: The HIV/AIDS epidemic in Cambodia has become a major problem in the last 7-8 years, mainly because in this formerly war-stricken country the socioeconomic situation is only slowly improving. Since only very few studies have been published to date on the oral health status of Cambodian HIV/AIDS patients, it was the purpose of the present investigation to study oral manifestations in Cambodian patients with HIV disease. METHODS: One hundred one Cambodian patients with HIV infection or AIDS were examined for the presence of oral manifestations in one medical center in Phomh Penh, Cambodia. RESULTS: Sixty three men and 38 women with a median age of 32 years were examined (age range 7.5 63.5 years). Of these patients, 42.6% were smokers, 46.5% of men were heavy drinkers and 90.5% of men were promiscuous compared with 5.3% of women. The most frequent AIDS-defining diseases were wasting syndrome (54.5%), Pneumocystis carinii pneumonia (PcP) (19.8%) and tuberculosis (18.8%). Puritic papular eruption, a common cutaneous manifestation in HIV-infected patients, was seen in 17.8% of patients. Candida-associated infections of the oral cavity were most common. Among the patients, 52.5% revealed pseudomembranous candidiasis and 35.6% had bilateral hairy leukoplakia. Only 10% of patients had no oral lesions. Also common were necrotising ulcerative gingivo-periodontal diseases (27.7%). CONCLUSION: The general health status of 101 Cambodian patients with HIV infection and AIDS was poor, and they demonstrated a large number of oral manifestations. Antiretroviral therapy is presently not available and only a fraction of patients receives antimycotic treatment (25.7%). HIV infection and the AIDS epidemic in Cambodia have become a serious problem and patients urgently need adequate diagnosis and antiretroviral therapies. PMID- 11896816 TI - Oral candidiasis as a clinical marker related to viral load, CD4 lymphocyte count and CD4 lymphocyte percentage in HIV-infected patients. AB - BACKGROUND: High viral load is currently considered to be one of the main indicators of the progression of HIV-induced immunodepression, but few studies have analysed its relationship to the presence of oral candidiasis (OC). The aim of this cross-sectional study is to analyse the relationship between viral load, total CD4 lymphocyte count, and percentage of CD4 lymphocytes to the occurrence of OC. METHODS: The present cross-sectional study included 156 HIV-infected patients seen at a clinic for sexually transmitted diseases and HIV. We assessed the presence or absence of OC, and microbiological samples were obtained from the palatine mucosa and dorsal tongue for a smear stained with KOH (potassium hydroxide) and culture on Sabouraud's dextrose agar in all patients. Viral load was determined by quantification of viral RNA in peripheral blood with a minimum detectable level of 500 RNA copies/ml. CD4+ counts/CD4+ percentage were categorized as <200/<14%, 200-499/14-28%, and >500/>29%, and HIV viral loads were categorized as <500, 500-10,000, >10,000 copies/ml. RESULTS: Thirty-eight percent (37.8%) of the patients had OC. Patients with CD4+ lymphocyte counts below 200 x 10(6)/l and CD4+ percentages below 14% showed a significantly higher frequency of OC (57.9% and 48.0%, respectively). Patients with a viral load over 10,000 copies/ml also had OC more frequently (44.8%). In the multiple logistic regression analysis, OC showed a statistically significant association with high viral load [>10,000 vs <500, odds ratio (OR)=11.4], low percentage of CD4+ lymphocytes (<14% vs >28%, OR=5), and injection drug use (IDU vs heterosexual transmission, OR=10.2). In HIV-infected patients, high viral load was associated with more frequent OC, regardless of CD4+ lymphocyte level. CONCLUSIONS: These findings suggest that oral candidiasis could be a useful clinical marker of patients with high viral load. In view of these results, emphasis should be placed on the importance of systematic examination of the oral cavity in all medical follow-up examinations of HIV-infected patients. PMID- 11896817 TI - The clinical relevance of epithelial dysplasia in the surgical margins of tongue and floor of mouth squamous cell carcinoma: an analysis of 37 patients. AB - BACKGROUND: The clinical relevance of the presence of epithelial dysplasia in the margins of surgically removed oral squamous cell carcinoma is still unclear. METHOD: In a retrospective study, the presence of mild or moderate epithelial dysplasia in the surgical margins of tongue and floor of mouth squamous cell carcinoma was examined histologically. Patients with tumor cells within 0.5 cm of the surgical margins were excluded. Also patients with severe dysplasia were excluded, as this is usually regarded as carcinoma in situ. Patients that received postoperative irradiation were also excluded. Only patients who completed a follow-up period of five years were included. All together, a total number of 37 patients fulfilled the inclusion criteria. RESULTS: Epithelial dysplasia was observed in 7 out of the 37 patients. Five of these patients, and two of the 30 patients with no dysplasia, had a local recurrence (P < 0.01). CONCLUSION: The presence of mild or moderate epithelial dysplasia in the margins of surgically removed oral squamous cell carcinoma carries a significant risk for the development of local recurrence. However, it should be noted that this study was of a retrospective nature and that the group of patients with epithelial dysplasia in the surgical margins was rather small. On the other hand, the inclusion criteria were somewhat strict, by limiting the oral subsite to tongue/floor of mouth, by excluding patients in whom tumors cell were found within 0.5 cm of the surgical margins and by excluding patients who received postoperative radiotherapy, amongst others. PMID- 11896818 TI - Prognostic role of p21WAF1 expression in areca quid chewing and smoking associated oral squamous cell carcinoma in Taiwan. AB - BACKGROUND: Alterations in p21WAF1 protein expression have been observed in a wide variety of human cancers by immunohistochemistry, and both decreased and increased levels of p21WAF1 protein expression have been shown to correlate with poor prognosis. METHOD: To examine the relation between p21WAF1 protein expression and prognosis in oral squamous cell carcinomas (SCCs), we performed an immunohistochemical study with antip21WAF1 antibody on 43 oral SCCs. Immunostaining results were then correlated with p53 protein levels, clinicopathological parameters of the tumors and overall patient survival. RESULTS: Of the 43 patients, 31 (72%) had tumors with positive p21WAF1 nuclear staining and 27 (63%) had tumors with p53 nuclear staining. There was no significant correlation between p21WAF1 and p53 protein expressions and both mutant p53-containing oral SCCs overexpressed p21WAF1 protein. In addition, no significant correlation was found between the p21WAF1 expression and the patients' age, sex, oral habit, cancer location, or primary tumor TNM status at the time of initial presentation. The Kaplan-Meier analysis showed a significant correlation between p21WAF1 protein overexpression and poor patient overall survival (P = 0.049). When p53 and p21WAF1 were evaluated together, the 5-year overall survival was lowest in p53(+)-p21WAF1(+) patients and highest in p53(-) p21WAF1(-) patients (P = 0.057). CONCLUSION: Combined evaluation of p21WAF1 and p53 expressions may be useful in estimating the prognosis of patients with oral SCCs in Taiwan. PMID- 11896819 TI - Intra-epithelial CD8+ T cells and basement membrane disruption in oral lichen planus. AB - BACKGROUND: We investigated basement membrane (BM) disruption and the distribution of mast cells (MCs) and T cell subsets, in oral lichen planus (OLP) and normal buccal mucosa (NBM) using immunohistochemistry. In OLP, there were increased numbers of tryptase+ MCs in areas of BM disruption (P < 0.05). METHOD: We identified clusters of intraepithelial CD8+ T cells in OLP, specifically in regions of BM disruption. The number of intraepithelial CD8+ T cells in regions of BM disruption was significantly greater than in regions of BM continuity (P < 0.05). RESULTS: There were comparable numbers of lamina propria CD8+ T cells in regions of BM disruption and BM continuity. The number of CD4+ T cells in the epithelium and lamina propria of OLP lesions did not vary between regions of BM disruption and BM continuity. CONCLUSION: These data suggest a role for MCs in epithelial BM disruption in OLP. CD8+ T cells may migrate through BM breaks to enter the OLP epithelium. PMID- 11896820 TI - Association between vascular endothelial growth factor (VEGF) expression and tumor angiogenesis in ameloblastomas. AB - BACKGROUND: Expression of vascular endothelial growth factor (VEGF), a major angiogenic factor, and microvessel density (MVD), assessed by the use of anti CD34 antibody, were immunohistochemically examined in benign and malignant ameloblastomas, as well as tooth germs, to clarify the possible role of angiogenesis in epithelial odontogenic tumors. METHODS: Specimens of 5 tooth germs, 35 benign ameloblastomas and 5 malignant ameloblastomas were examined by immunohistochemistry using anti-VEGF and CD34 monoclonal antibodies. RESULTS: Immunoreactivity for VEGF was detected in both normal and neoplastic odontogenic epithelial cells, and weakly in microvessels near odontogenic epithelial cells, suggesting that this angiogenic factor acts on endothelial cells via a paracrine mechanism in odontogenic tissues. Both benign and malignant ameloblastomas showed elevated VEGF expression as compared to tooth germs. VEGF expression was low in keratinizing cells in acanthomatous ameloblastomas and granular cells in granular cell ameloblastomas, and acanthomatous ameloblastomas showed the lowest VEGF reactivity among the subtypes of ameloblastomas. MVD in both benign and malignant ameloblastomas was higher than that in tooth germs, indicating increased demands for blood in the neoplastic tissues. CD34-positive microvessels in follicular ameloblastomas were numerous and small, whereas those in plexiform ameloblastomas were scattered and dilated. MVD tended to depend on VEGF expression levels in both benign and malignant ameloblastomas. CONCLUSIONS: VEGF was considered to be an important mediator of angiogenesis in these epithelial odontogenic tumors, and up-regulation of VEGF might be associated with neoplastic or malignant changes of odontogenic epithelial cells. PMID- 11896821 TI - Adhesion of a chondrocytic cell line (USAC) to fibronectin and its regulation by proteoglycan. AB - BACKGROUND: Chondrocytes produce various extracellular matrices during chondrogenesis. Fibronectin and proteoglycan are major extracellular matrix proteins in cartilage tissue, but the interactions between them are not clear. METHODS: Recently, we succeeded in establishing a cell line (USAC) with phenotypes of chondrocytes from a human osteogenic sarcoma of the mandible. Using this cell line, cell adhesion to fibronectin, the effect of proteoglycan on the cell adhesion and expression of integrin alpha5beta1 were investigated. RESULTS: Cells immediately adhered to fibronectin and then spread. Proteoglycan inhibited cell adhesion to fibronectin dose-dependently, whereas collagen did not. The expression of both mRNAs of alpha5 and beta1 subunits was detected 12 h after treatment with proteoglycan, but the expression of beta1 subunit mRNA had diminished by 24 h after treatment. CONCLUSIONS: These findings suggest that proteoglycan might modulate signal transduction from fibronectin by decreasing the expression of alpha5beta1 integrin. PMID- 11896822 TI - Immunohistochemical study of interleukin-1beta and interleukin-1 receptor antagonist in an antigen-induced arthritis of the rabbit temporomandibular joint. AB - BACKGROUND: In temporomandibular joint (TMJ) arthritis, there is limited knowledge of the relationship between interleukin-1beta (IL-1beta) and interleukin-1 receptor antagonist (IL-1ra), as well as the source of these cytokines. We investigated the development of an antigen-induced arthritis in the rabbit TMJ immunohistochemically. METHODS: Unilateral TMJ arthritis was induced in 32 adult New Zealand White rabbits. From 6 h to 12 weeks after induction of arthritis, topology of IL-1beta and IL-1ra were observed. RESULT: The acute stage of induced arthritis lasted for one week after induction, thereafter it became chronic. In the early phase of the acute stage, infiltrating inflammatory cells, as well as synovial cells, produced IL-1beta and IL-1ra. In the late phase of the acute stage, the main source of these cytokines was subsynovial fibroblasts. In this phase of arthritis, IL-1beta and IL-1ra did not appear to be produced by synovial cells. From the early to intermediate phase of the chronic stage, proliferating synovial cells produced IL-1beta and IL-1ra. In this phase of the arthritis, these cytokines were also observed in a cluster formation in chondrocytes. CONCLUSION: This arthritis model shows a staging of the joint inflammation process with time. IL-1beta and IL-1ra are produced by a certain kind of cells depending on the stage of inflammation. PMID- 11896823 TI - Gingival fine needle aspiration cytology in acute leukemia. AB - BACKGROUND: Oral findings in acute leukemia (AL) are common and could be the presenting feature of the disease, namely, gingival enlargement, ulceration, bleeding, and infection. Gingival enlargement in AL is either due to leukemic infiltration, or due to reactive hyperplasia. To differentiate between them a biopsy is required, but being highly contraindicated, biopsy has been substituted in this study by fine needle aspiration cytology (FNAC). METHOD: Gingival FNAC was performed on different sites in the upper and lower gingiva. Each site represents an interdental papilla, which was selected according to the clinical presentation, i.e., being enlarged or not. Seventy-two adult AL patients received a cytological and clinical examination in this study, and the cases were classified and categorized according to the French-American-British (FAB) criteria. RESULTS: Twenty-one cases were diagnosed as being infiltrated, 16 with gingival enlargement, 4 with no evidence of enlargement. In one case the infiltration affected the alveolar mucosa of an edentulous patient. In six cases the ginigva was enlarged without being infiltrated (reactive hyperplasia). Leukemic gingival enlargement was seen mostly in patients with acute myeloid leukemia, particularly M4 and M5 subtypes; however, two patients with acute mixed lineage leukemia (AMLL) were both affected with leukemic infiltration. CONCLUSION: FNAC was shown to be a simple, non-traumatic and useful diagnostic procedure for screening leukemic infiltration in gingival tissues in AL patients. PMID- 11896824 TI - Expression of ICAM-1 and E-selectin in gingival tissues of smokers and non smokers with periodontitis. AB - BACKGROUND: Tobacco smoking affects systemic concentrations of soluble intercellular adhesion molecule (ICAM)-1, but its effect on local expression of adhesion molecules in gingival tissue has not been studied previously. METHODS: E selectin and ICAM-1 expression on small blood vessel endothelia in gingival biopsies obtained from smokers (n=17) and non-smokers (n=17) with periodontitis was examined with immunohistochemistry. Blood vessels were identified with monoclonal antibody for von Willebrand's factor. RESULTS: A significantly larger number of vessels were observed in inflamed tissues of non-smokers than smokers (P<0.05). The number and proportion of vessels expressing both ICAM-1 and E selectin was greater in sites with inflammation compared to non-inflamed sites in both smokers and non-smokers (P<0.05). The proportion of the total number of vessels expressing ICAM-1 in non-inflamed sites was greater in non-smokers compared with smokers (P<0.05). CONCLUSIONS: These results suggest that the inflammatory response in smokers with periodontitis may not be accompanied by an equivalent increase in vascularity. Reduced ICAM-1 expression in non-inflamed areas of smokers could reflect a systemic effect of tobacco smoking on ICAM-1 independent of inflammation. PMID- 11896825 TI - ABO blood group antigens in oral mucosa. What is new? AB - Histo-blood group ABH (O) antigens are major alloantigens in humans. These antigens are widely distributed in human tissues and undergo changes in expression during cellular differentiation and malignant development. The ABH antigens have been characterized as terminal disaccharide determinants which represent secondary gene products. They are synthesized in a stepwise fashion from a precursor by the action of different glycosyltransferases. In non keratinized oral mucosa, a sequential elongation of the carbohydrates is associated with differentiation of epithelial cells, resulting in expression of precursors on basal cells and A/B antigens on spinous cells. Reduction or complete deletion of A/B antigen expression in oral carcinomas has been reported, a phenotypic change that is correlated with invasive and metastatic potential of the tumours and with the mortality rates of the patients. Disappearance of the antigens is ascribed to the absence of A or B transferase gene expression. Several studies have shown that loss of A and B antigen expression is associated with increased cell motility, invasion in matrigel, and tumourigenecity in syngenic animals. In vivo studies of human oral wound healing show similarly decreased expression of A/B antigens on migrating epithelial cells. Some studies suggest that the relationship between expression of blood group antigens and cell motility can be explained by different degrees of glycosylation of integrins. Changes in ABO expression in tumours have, in some cases, been due to the A/B gene promoter, although little is known about the regulation of A, and B expression, in normal tissue. PMID- 11896826 TI - Antioxidant enzyme levels in oral squamous cell carcinoma and normal human oral epithelium. AB - BACKGROUND: The antioxidant enzymes (manganese- and copper-zinc-containing superoxide dismutases, catalase and glutathione peroxidase) limit cell injury induced by reactive oxygen species. The purpose of the study was to determine whether human oral squamous cell carcinomas have altered antioxidant enzyme levels. This study is the first to undertake this task in human oral mucosa and squamous cell carcinoma. METHODS: Semiquantitative immunohistochemistry was used to examine 26 archived oral squamous cell carcinoma biopsies. Fourteen well differentiated and 12 poorly differentiated tumors were examined, as were 12 specimens of oral mucosa. All sections were reviewed by two oral and maxillofacial pathologists, and image analysis of the immunostained sections was performed using NIH Image. Antioxidant enzyme staining intensities were compared in the different groups by Duncan's multiple range test. RESULTS: In general, mucosal basal cells displayed lower antioxidant enzyme levels than spinous cells, and primary tumor cells displayed lower antioxidant enzyme staining intensities than did their normal cell counterparts. Moreover, poorly differentiated tumor cells showed lower antioxidant enzyme staining intensities than well differentiated tumor cells. Manganese-containing superoxide dismutase staining intensities were, however, higher in well-differentiated oral squamous cell carcinomas than their normal cells of origin. CONCLUSIONS: Detection of antioxidant enzymes may be a useful future marker in the molecular diagnosis of the oral cancer. Moreover, it may be possible to not only monitor the effectiveness of chemopreventive and therapeutic strategies in oral cancer using these enzymes, but to monitor tumor recurrence. PMID- 11896827 TI - Immunolocalization of c-Fos and c-Jun in human oral mucosa and in oral squamous cell carcinoma. AB - BACKGROUND: Studies have addressed the relevance of c-Jun and c-Fos proteins in cancer development. In the present study, the expression of c-Jun and c-Fos, the major components of transcription factor activator protein (AP1), were evaluated to determine possible alterations to these factors in oral squamous cell carcinoma (OSCC). METHODS: Fifteen cases of normal oral mucosa and 20 cases of OSCC were retrieved from the Archives of the Surgical Pathology Service at the University of Sao Paulo. The samples of normal oral mucosa or OSCC originated from different oral mucosal sites. Tissues were submitted for immunohistochemical analysis to detect c-Jun and c-Fos proteins. The OSCC was classified as well, intermediate or poorly differentiated. RESULTS: The results showed that both c Jun and c-Fos are expressed in normal oral mucosa and in OSCC. In normal mucosa, immunoreactivity for c-Jun was detected in the cytoplasm of the upper basal layers, while in OSCC, c-Jun was detected in the nuclei of the cells. C-Fos expression was observed in the nuclei of cells, both in normal mucosa and in OSCC, but its expression varied according to the cell layer in normal mucosa, and the differentiation of OSCC. CONCLUSIONS: The nuclear expression of c-Jun in OSCC, in contrast to its cytoplasmic expression in normal oral mucosa, indicates that c-Jun may have a role in the development of oral cancer. In contrast, the absence of both c-Jun and c-Fos in poorly differentiated carcinoma might be useful in understanding the cell cycle events important in uncontrolled cell growth. PMID- 11896828 TI - The mRNA expression of inducible nitric oxide synthase in DMBA-induced hamster buccal-pouch carcinomas using reverse transcription-polymerase chain reaction. AB - BACKGROUND: Three isoforms of NO synthase (NOS) have been identified: endothelial NOS, neuronal NOS, and inducible NOS (iNOS). The enhanced expression of iNOS, at the protein level, has been reported previously in certain chemically induced oral carcinomas in hamster buccal-pouch mucosa, however, the corresponding expression of iNOS, at the mRNA level, has not yet been demonstrated. The purpose of the present study is to assess the iNOS mRNA expression level in 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster buccal-pouch carcinomas using a reverse transcription-polymerase chain reaction (RT-PCR). METHODS: Thirty-three outbred, young (six-weeks old), male, Syrian golden hamsters (Mesocricetus auratus) were randomly divided into one experimental group (13 animals) and two control groups (10 animals each). The pouches of a group of 13 animals of the experimental group were bilaterally painted with a 0.5% DMBA solution three times a week for 12 weeks. Each animal of one control group was similarly treated with mineral oil only, while the other control group of 10 animals remained untreated throughout the experiment. RESULTS: Areas of dysplasia and squamous-cell carcinomas, with a 100% tumor incidence, developed for all of the DMBA-treated buccal pouches. The mineral oil-treated and untreated pouches had no obvious changes. A band of 499-bp, corresponding to iNOS mRNA, was present in all the DMBA-treated hamster buccal-pouch mucosa animals, but not in the untreated animals or the animals treated with mineral oil. Upon direct sequencing, this 499 bp band was confirmed to be part of the iNOS gene. CONCLUSIONS: This study demonstrated that increased iNOS mRNA expression could contribute to the mechanism for experimentally induced oral carcinogenesis. PMID- 11896829 TI - Cytokeratin expression patterns in jaw cyst linings with metaplastic epithelium. AB - BACKGROUND: Cytokeratin (CK) expression patterns have been studied in numerous intact and diseased oral tissues. However, CK expression in metaplastic squamous cells has not been explored in depth and the origin of metaplastic epithelial linings of the jaw cysts has not been sufficiently investigated. METHODS: We examined CK expression in 46 postoperative maxillary cysts (POMCs) which were lined with pseudostratified columnar cells only, columnar and squamous cells, and squamous cells only, in 13, 30 and 3 cases, respectively. RESULTS: The expression of CK8, CK13 and CK18 were observed in 39, 9 and all 43 of the columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13, and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The patterns of expression of CK13 and CK18 observed were CK18(+)-CK13(-) in 10 metaplastic linings, CK18(+)-CK13(+) in 16, and CK18(-)-CK13(+) in 7. The expression of CK13- and CK18-mRNA was generally correlated with level of protein expressed. CK18-mRNA expression was observed by in situ hybridization, not only in the 26 metaplastic linings which were positive for CK18 protein, but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings, although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed inversely to the CK18-mRNA. CONCLUSIONS: These results indicate that CK18-mRNA is preserved through metaplasia, although the protein expression decreased. Metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression. PMID- 11896830 TI - Interobserver and intraobserver variability in the clinical assessment of oral lichen planus. AB - BACKGROUND: In 1978, a clinical definition of OLP was formulated by the WHO. To date, the validation results of this clinical definition have not been published. The aim of this study was to evaluate interobserver and intraobserver variability in the clinical assessment of oral lichen planus (OLP). METHODS: Four clinicians examined a set of 159 clinical pictures of a white lesion in a group of 60 patients. Each reviewing examiner was asked to apply the WHO definition of OLP from 1978, and to categorise each case as either: (i) diagnostic of OLP, (ii) other definable lesion, or (iii) leukoplakia. After three months, each of the four reviewing clinicians was given the clinical pictures of 45 randomly retrieved cases from the original 60. Interobserver and intraobserver variability were assessed by calculation of unweighted kappa statistics. RESULTS: Interobserver agreement varied from 0.43 (moderate) to 0.77 (substantial), while the intraobserver agreement varied from 0.62 (substantial) to 0.92 (good). CONCLUSIONS: Although the clinical WHO definition of OLP seems to be more reproducible than the histopathological one, there is still a significant amount of subjectivity in using this definition. A set of clinical and histopathological diagnostic criteria with good interobserver and intraobserver agreements (kappa values > 0.8) is very important in enabling reproducible and reliable studies on OLP to be performed. PMID- 11896831 TI - Prevalence of fluconazole-resistant strains of Candida albicans in otherwise healthy outpatients. AB - BACKGROUND: In contrast to the immunosuppressed patient population, the prevalence of fluconazole-resistant strains of Candida albicans among healthy individuals has not been extensively studied. METHODS: Candida species were cultured form 50 healthy outpatients with clinical signs of oral candidiasis. Following one week of the recommended fluconazole regimen, post-treatment cultures were obtained. Both pre- and post-treatment yeasts were identified and in vitro susceptibility testing was performed using the NCCLS M-27A method. Strains were further differentiated using established cDNA probes. RESULTS: Forty four patients (88%) had positive C.albicans cultures prior to treatment. Antifungal susceptibility testing of these strains demonstrated no in vitro resistance to fluconazole. At post-treatment evaluation, eight patients (18%) had persistent signs of infection and 10 patients (23%) had positive Candida sp. cultures despite no clinical signs of infection. DNA analysis confirmed that the same C. albicans strain was present both in the pre-treatment and the post treatment cultures. CONCLUSIONS: Our results showed that the presence of fluconazole-resistant strains of C.albicans does not appear to be prevalent among healthy outpatients furthermore, in vitro antifungal susceptibility testing does not always predict successful therapy in these patients. PMID- 11896832 TI - Salivary levels of vascular endothelial growth factor (VEGF) in recurrent aphthous ulceration. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a multifunctional angiogenic cytokine involved in angiogenesis and wound healing. Its presence in recurrent aphthous ulceration has not been reported to date. The aim of this study was to investigate the association of salivary levels of VEGF with various stages of recurrent aphthous ulceration (RAU). METHODS: VEGF levels were determined in a group of 27 age and sex-matched healthy controls and in 30 patients with minor and major RAU grouped into the three stages: (I) early active stage, (II) active stage, and (III) remission period. VEGF levels (pg/ml; mean +/ SD) in unstimulated whole saliva were determined by enzyme immunoassay. RESULTS: Patients with major RAU - stages I and II - had decreased VEGF values (765 +/- 458 and 341 +/- 109, respectively) when compared both to healthy controls (1652 +/- 567; P < 0.01) and to stage III major RAU (1524 +/- 784; P < 0.005). CONCLUSION: Salivary VEGF levels seemed to be associated with ulcer development in major RAU, showing stage-dependent alterations during the course of this disorder. PMID- 11896833 TI - The excretion of cephem antibiotics into saliva is inversely associated with their plasma protein-binding activities. AB - BACKGROUND: The excretion of medicated drugs into saliva may disturb the oral environment and antibiotic excretion into saliva appears to be regulated by many factors that have not been fully explored. METHODS: Excretion of four cephem antibiotics into saliva was examined in healthy volunteers and rats, using high performance liquid chromatography, and the relationship between excretion levels and plasma protein-binding activities of the antibiotics was investigated. RESULTS: Following addition of 50 microgram/ml of each antibiotic to human plasma, protein binding rates (PBRs) of cefuzonam (CZON, molecular weight (MW): 535.58), cefotaxime (CTX, MW: 477.45), flomoxef (FMOX, MW: 518.45) and cefozopran (CZOP, MW: 551.99) were 87.8 +/- 1.2, 70.8 +/- 0.8, 36.2 +/- 0.5 and 8.3 +/- 0.3%, respectively. In rat plasma, PBRs of the four antibiotics were 94.0 +/- 0.5, 62.1 +/- 1.4, 54.0 +/- 0.8 and 6.0 +/- 0.8%, respectively. Similar PBRs were observed when the antibiotic concentration was increased to 100 and 200 microgram/ml. CZOP was most rapidly excreted into saliva and had the highest concentration in saliva among the tested antibiotics, while the plateau level of CZON was the lowest. The excreted levels of each antibiotic in saliva, when locally perfused through the rat facial artery, were inversely associated with each PBR. Similarly, the ratios of antibiotic concentration in saliva to rat plasma were almost constant for each antibiotic, revealing an inverse relationship with PBRs. CONCLUSION: These results appear to indicate that low molecular weight antibiotics are excreted into saliva through passive diffusion, inversely relating to their PBRs, and that high concentrations of antibiotics in the saliva have the potential to change the oral ecological environment. PMID- 11896834 TI - Whole stimulated salivary flow in patients with chronic hepatitis C virus infection. AB - BACKGROUND: Salivary gland disorders have been included among the extra-hepatic manifestations related to HCV infection. METHODS: The whole stimulated salivary flow rate (spitting technique) was studied in 74 HCV infected patients to evaluate salivary gland dysfunction. RESULTS: The salivary flow of the patients with chronic HCV infection was similar to that of the healthy controls. The association between subjective xerostomia salivary flow was seen to be very weak. No significant associations were found between salivary flow and age, sex, risk factor of acquired infection, ALT, AST, GGT, ALP values, time lapsed since the diagnosis or HCV-RNA detection in saliva. CONCLUSIONS: Although the functional repercussion of hepatitis C related lymphocytic sialoadenitis remains unclear, we did not find a significant reduction in the whole stimulated salivary flow in HCV infected patients. PMID- 11896835 TI - Oral pemphigus vulgaris occurring during pregnancy. AB - There have been few reports describing the occurrence of pemphigus vulgaris (PV) during pregnancy. The patient described in this case report is interesting because the PV that developed during her pregnancy was confined to her mouth. It has been suggested that prompt treatment with systemic steroids prevents development of PV in cutaneous tissues. In this case, early control of the condition is believed to have eliminated the need for high dose steroids throughout the remainder of the pregnancy. In addition, this therapeutic approach could have contributed to the birth of a baby free of PV. Resolution of the presenting oral symptoms allowed the mother to resume a normal diet, allaying her anxiety about the possible effects of poor nutritional intake on foetal development. Aspects of clinical management considered in this report include the choice of immunosuppressive therapy and the multidisciplinary care involving both dental and obstetric specialists. PMID- 11896836 TI - Consequences of oral rehabilitation on dyskinesia in adults with Down's syndrome: a clinical report. AB - The aim of this article is to demonstrate that the presence of orofacial dyskinesia is often owing to underlying facial dysmorphology in persons with Down's syndrome. A series of cases is presented where orofacial dyskinesia was successfully treated by therapy establishing occlusal stability. The diagnosis of dyskinesia owing to dysmorphology should be precluded before any link with the degree of intellectual disability or neurological deficit is presumed. A multidisciplinary approach may be necessary to diagnose and treat these patients. PMID- 11896837 TI - Dentists' perceptions of dentine hypersensitivity and knowledge of its treatment. AB - The aim of the present investigation was to determine by questionnaire, UK dentists' perception of Dentine Hypersensitivity (DH) and knowledge of its treatment. A total of 403 questionnaires were sent to a selected group of UK dentists who had either inquired about further postgraduate education or had attended a course at the Eastman Dental Institute/Hospital. A total of 181 of 403 dentists (44.9%) (118M; 36F, 17 no response, mean age 38.2 years [s.d. 8.97]) returned the questionnaire. About 92.8% (n=168) of responding dentists claimed to see patients with DH in their practice. According to the dentists' replies at least one of four of their patients suffered from the complaint. About 71.8% (n=130) of dentists reported that DH was a severe problem in at least 10% of their patients and that pain from DH lasted no more than 4 weeks. Most of responding dentists claimed to be asked about DH by their patients and stated that they offered advice or treatment to their patients. Nearly 87.3% (n=158) of responding dentists provided a wide range of treatment options/advice which included both in-office and over-the-counter (OTC) products. Popular responses included desensitizing pastes/gels, Topical F(-) varnishes and toothpastes/rinses/gels, advice on atraumatic tooth brushing, dentine bonding agents (DBA), glass-ionomer cements (GIC) and other unspecified restorations. Of the various in-office treatments Duraphat was the most cited choice of varnish/primer options. Sensodyne toothpaste was the most popular of the specified OTC products. Most dentists appeared to understand the aetiological features associated with DH and provided a wide range of factors including the effects of incorrect tooth brushing, dietary acids as well as the possible influence on non-dental topics such as stress. Most responding dentists believed that their advice on DH was generally effective although they did highlight that certain aspects on the condition were lacking such as appropriate scientific information including the prevention of DH and its treatment. The results from the present study highlight several discrepancies in the perception and knowledge of the treatment of DH between dentists and their patients. The results from this study are, however, reasonably consistent with those previously reported by Dutch investigators. The results from this study also highlighted a need for guidelines on the aetiology, prevention and treatment of DH for both dentists and their patients. The reported average frequency and duration of discomfort from DH by the responding dentists appeared to be consistent with the available literature. PMID- 11896838 TI - Dentine hypersensitivity in subjects recruited for clinical trials: clinical evaluation, prevalence and intra-oral distribution. AB - Relatively few studies have reported on the frequency, distribution and severity of dentine hypersensitivity (DH) in subjects recruited for clinical trials of desensitizing agents. Potential subjects (n= 48 M, 81 F, mean age 35.1 years) for inclusion into such a study were screened to determine the extent of the problem. 117 subjects (41 M, 76 F) mean age 24.9 years were clinically examined. Evaluation by questionnaire indicated that the prevalence of DH was proportionately higher in the 20-29.9 years (34.9%), and 30-39.9 years groups (33.3%), respectively. Sensitivity to cold was the main presenting symptom. Tactile (probe) and cold air (dental air syringe) stimuli were used to clinically evaluate DH. Of the teeth eligible for evaluation 1561/3136 (49.8%) responded to either one or both of the test stimuli; 274/3136 (8.7%) responded to tactile only stimulation, 779/3136 (24.8%) to thermal only stimulation and 508/3136 (16.2%) to both tactile and thermal stimulation. Of those teeth responding to the stimuli, 477 (30.6%) were premolars, 437 (28%) incisors, 415 (26.8%) molars and 232 (14.9%) canines. The results agree with those of previously reported studies in that DH is most frequently observed on premolars and that proportionately more teeth are sensitive to evaporative than to tactile stimulation. Furthermore it would appear from the results of the study that tactile is less effective than thermal/evaporative stimulation in the evaluation of DH. PMID- 11896839 TI - Influence of removable partial denture on periodontal indices and microbiological status. AB - Thirty patients (19 men and 11 women) were provided with a removable partial denture (RPD) and assigned randomly to two groups: 15 patients were called back twice a year for plaque control, reinforcement of instructions, denture hygiene control and professional prophylaxis; the other 15 were not called back. The 30 patients were examined after 2-3 weeks following the end of the prosthetic treatment, after 1 and 2 years. At each examination, the following parameters were recorded [gingival inflammation, plaque index (Pl I), tooth mobility, attachment level, pocket depth] and a bacteriological examination of subgingival plaque was carried out. Few differences appeared between the two groups; the values observed show a relatively low level of hygiene and but little motivation with regard to prophylaxis techniques. PMID- 11896840 TI - Severe tooth loss among UK adults--who goes for oral rehabilitation? AB - A random probability sample of 2667 United Kingdom (UK) addresses was selected in a multistage sampling process. Participants were interviewed about their oral health status--number of teeth possessed and denture status. In addition information was collected about their socio-demographic characteristics (age, gender, social class and income level) and dental service factors--type of service used and difficulty accessing national health service (NHS) care. The response rate was 70%. Six percent (107) claimed they had less than 20 teeth but did not use a denture. Variations in this practice were apparent in relation to a number of socio-demographic factors: age (P < 0.05), gender (P < 0.01) and social class (P < 0.001) but not service related factors. In regression analysis, social class, gender and age emerged as important predictors of this practice. For example, those from lower social classes were approximately half as likely to use dentures despite experiencing considerable tooth loss (OR=0.53, 95% CI 0.34, 0.83), having controlling for other factors. More than one in 20, in Britain claim they have experienced considerable tooth loss but are without resource to a denture. Socio-demographic factors rather than service related factors are associated with this practice, particularly social class. PMID- 11896841 TI - Relationship between the flow of bolus and occlusal condition during mastication- computer simulation based on the measurement of characteristics of the bolus. AB - The purpose of the present study was to clarify the relationship between the flow of a bolus and occlusal condition during mastication. First, the characteristics of a bolus under mastication was measured in subjects having different occlusal conditions. Secondly, the flow of a bolus between the upper and lower first molars under mastication was simulated using finite element non-linear dynamic analysis. Measurement of the elasticity of the bolus clarified the phenomenon of its communition. The measurement of the viscosity of the bolus clarified the phenomenon of its mixing with saliva. In addition, a relationship between the elasticity and the viscosity of the bolus at the point of just before swallowing was investigated. The flow of the bolus under mastication was revealed to vary according to the occlusal condition. These results suggest a close relationship between the occlusal condition, the flow of the bolus and its characteristics. PMID- 11896843 TI - An investigation into the transverse and impact strength of "high strength" denture base acrylic resins. AB - A range of materials, often marketed as high strength resins is available. These materials are often expensive options to conventional heat-cured acrylic resin. The aim of this study was to investigate transverse and impact strength of five "high strength" acrylic resin denture base materials. A conventional heat-cured acrylic resin was used as a control. Specimens were prepared as specified in the International Standard Organization (ISO 1567: 1988) and British standards for the Testing of Denture Base Resins (BS 2487: 1989) and the British Standard Specification for Orthodontic resins (BS 6747: 1987) for transverse bend and impact testing. The impact strength was measured using a Zwick pendulum impact tester and the transverse bend strength measured using a Lloyds Instruments testing machine. The results showed that Metrocryl Hi, Luctitone 199 and N.D.S. Hi all had an impact strength which was significantly higher than the control. For the modulus of rupture, there was a significant difference between Sledgehammer and the other groups. There was no significant difference between the other groups and the control. For the modulus of elasticity, Sledgehammer produced the highest value followed by the control. The remaining four materials had a modulus of elasticity less than the control. PMID- 11896842 TI - Mechanical properties of resin cements with different activation modes. AB - Dual-cured cements have been studied in terms of the hardness or degree of conversion achieved with different curing modes. However, little emphasis is given to the influence of the curing method on other mechanical properties. This study investigated the flexural strength, flexural modulus and hardness of four proprietary resin cements. Materials tested were: Enforce and Variolink II (light , self- and dual-cured), RelyX ARC (self- and dual-cured) and C & B (self-cured). Specimens were fractured using a three-point bending test. Pre-failure loads corresponding to specific displacements of the cross-head were used for flexural modulus calculation. Knoop hardness (KHN) was measured on fragments obtained after the flexural test. Tests were performed after 24 h storage at 37 degrees C. RelyX ARC dual-cured showed higher flexural strength than the other groups. RelyX ARC and Variolink II depended upon photo-activation to achieve higher hardness values. Enforce showed similar hardness for dual- and self-curing modes. No correlation was found between flexural strength and hardness, indicating that other factors besides the degree of cure (e.g. filler content and monomer type) affect the flexural strength of composites. No statistical difference was detected in the flexural modulus among the different groups. PMID- 11896844 TI - A physiotherapeutic approach to craniomandibular disorders: a case report. AB - This is a case report of a 19-year-old female who presented with a unilateral weakness of the right masseter muscle evidenced by electromyographic examination. The presence of a mandibular deviation to the right during opening because of this weakness was treated with neuromuscular electrical stimulation (NMES). After the physiotherapeutic treatment, the electrical activity of the right masseter muscle increased during function and the mandibular deviation disappeared. Electromyography (EMG) can have a useful role in the determination of the muscular profile, and for evaluating therapeutics. PMID- 11896845 TI - An index for analysing the stability of lateral excursions. AB - Five lateral excursions on both left and right sides of 20 healthy subjects were recorded by Gnatho-hexagraph, which was an opto-electronic system with six degrees of freedom. The incisor point, the working condylar point (WCP) and the balancing condylar point (BCP) were analysed. When the incisor point moved 1, 2, 3, 4, 5 mm on its path, the incisor point and corresponding condylar points were analysed. The linear distances from the intercuspal position (ICP) to each analysed point at each analysed position were calculated as Linear distance. The distance between five lateral excursions in x, y, z, space coordinates at each analysed position of three analysed points were calculated as Dx, y, z, s. To evaluate the stability of lateral excursions, the Index S was defined as the equation (S=D/Linear distance * 100) and calculated. From the result of Index S, the WCP showed significantly less stability than the other two analysed points (P < 0.05). The incisor point showed best stability, especially in upper-lower direction. The BCP also showed good stability. Index S revealed the characteristic movement in x, y, z, space coordinates at each analysed point and it showed clearly the stability of lateral excursions. PMID- 11896846 TI - Immunolocalization of vimentin and alpha-smooth muscle actin in dysfunctional human temporomandibular joint disc samples. AB - The expression of vimentin and alpha-smooth muscle (alpha-SM) actin was examined in 10 human temporomandibular joint (TMJ) disc samples, with internal derangement and in two control specimens, in order to evaluate the phenotypic characteristics of TMJ disc cells in relationship to histological findings. This was accomplished by means of monoclonal antibodies specific for vimentin and alpha-SM actin and immunocytochemical technique. The study, revealed that every disc cell constantly expressed vimentin. Scattered alpha-SM actin positive cells could be appreciated in normal TMJ discs and tissues with minor pathological findings. In TMJ discs with severe alterations, i.e. tears and clefts, almost fibroblast-like cells, fibrochondrocytes and chondrocyte-like cells were strongly immunolabelled by anti alpha-SM actin antibody. According to these findings it can be assumed that vimentin is expressed by all disc cell populations and it appears not to be influenced by any disease condition of the disc; on the other hand the up regulation alpha-SM actin immunolabelling seems to be correlated to histopathological findings of tears and clefts. Cells, with a contractile phenotype, close to such defects, could be involved in disc tissue contraction and repair. The plasticity of disc cell populations which evolve towards a different phenotype when subjected to action of macro- and micro-environmental factors is also supported. PMID- 11896847 TI - Effect of calcium supplementation on bone dynamics of the maxilla, mandible and proximal tibia in experimental osteoporosis. AB - The effect of calcium supplementation on the bone dynamics in the hard palate of the molar region (maxilla), mandible and proximal tibia in experimental osteoporotic rats was examined. Ninety ovariectomized (OVX) and 45 sham-OVX Wistar female rats were used in this study. All the rats received surgical operation at 6 weeks of age. Ovariectomized rats were fed on a low calcium diet (0.02%) for 12 weeks post-operation, and then randomly divided into the two following groups. One group was fed on high calcium diet (2.30%) (OVX-HCa) and the other group was remained on the low calcium diet (OVX-LCa). Sham-OVX rats were fed on regular calcium diet (1.15%) during the experimental period (Sham OVX). Histomorphological analysis was carried out from 12 to 32 weeks post operation. On undecalcified thin section, bone volume, eroded surface, osteoid surface and bone formation rate were calculated for cortical bone of the maxilla, and for cancellous bone of the mandible and proximal tibia. In the OVX-LCa group, compared with the Sham-OVX group, decrease of the bone volume and increase of the bone resorption and formation parameters were detected throughout the observation periods. In the OVX-HCa group, compared with the OVX-LCa group, increase of the bone volume and temporarily increased parameters of bone formation at 1 week after feeding on high calcium diet were observed in the maxilla, but these changes were not observed in the mandible and proximal tibia. Moreover, the bone resorption and formation parameters in the maxilla, mandible and proximal tibia in the OVX-HCa group became equivalent to the Sham-OVX levels with the passage of time. PMID- 11896848 TI - A survey of amalgam restorations in a south-western Nigerian population. AB - The present survey assessed the proportion of replacement restorations in comparison with new restorations because of primary caries in a setting where the caries experience had been reported to be low but probably on the increase. Also the relative importance of the main reasons for the replacements among other things was established. A total of 488 amalgam restorations were surveyed. About 25% of all restorations were replacements of failed restorations. The main reason for the replacement was bulk amalgam fracture, which accounted for 47.1% of all restorations. These show a reversal of what had been reported in settings where the caries experience was high but now is on the decrease. The importance of bitewing radiograph of all failed restorations was emphasized in order to minimize under or over treatment. The median age of failed restorations was 5.0 years. More than half (60.3%) of replaced restorations had failed during the first 5 years of use. PMID- 11896849 TI - Oral hygiene habits, denture cleanliness, presence of yeasts and stomatitis in elderly people. AB - The purpose of this study was to determine oral hygiene habits, denture cleanliness, presence of yeasts and denture stomatitis in elderly people. Seventy complete denture wearers were investigated clinically and mycologically. Subjects were evaluated according to, presence of denture stomatitis, presence of yeasts, denture cleanliness, frequency of denture brushing and denture cleaning methods. Swabs were taken from the palate investigated mycologically in order to identify the yeast colonies. No statistical relationship was found between denture stomatitis and frequency of denture brushing and denture cleaning methods. However, there was a statistically significant relationship between denture stomatitis, yeasts' presence and denture cleanliness. PMID- 11896850 TI - The 1990s: a time for change and opportunity in Finnish mental health services. PMID- 11896851 TI - Integration of theory and practice in learning mental health nursing. AB - This article describes an action research project that aimed at a better integration of theory and practice in the education of mental health nursing students. Two partners, an institute of nursing and health care and a university hospital, collaborated to develop a new educational programme for mental health nursing. The blocks of theoretical studies were implemented simultaneously with practical training, and the theory content was taught by nursing teachers as well as by nurse practitioners who worked on the teaching wards. In addition, the students had their own personal nurse-preceptors on the wards. The nurse managers were responsible for the educational level of the teaching wards and the director of nursing planned the teaching arrangements together with the nursing teachers. In all, the project involved over 50 different actors and several researchers. The results are encouraging: all the participants - students, preceptors, nurse managers and nursing teachers - found the project rewarding and they want to continue to develop and improve the level of teaching and learning in mental health nursing education. All the participants grew and developed professionally during the project. PMID- 11896852 TI - The resources of parents with a child in psychiatric inpatient care. AB - The purpose of this study was to analyse and describe the resources of parents with a child in psychiatric inpatient care. The factors associated with parental coping were also assessed. The data were collected from 19 hospitals with a child psychiatry unit. At the time of data collection, all the parents of the children in psychiatric inpatient care in these hospitals were invited to participate in the study (N = 268). The method of data collection was a questionnaire including open-ended questions. The response percentage was 30% (N = 79). The data were analysed using SPSS software. The replies to the open-ended questions were analysed utilizing data-based content analysis. The findings indicated that the parents received more emotional than instrumental support. Few got financial support. The parents got support from their spouses, families, friends and fellow workers and the health care personnel. Nearly half of the parents wanted more support from health care personnel. Of the different kinds of social support reported, emotional support, support in the care and rearing of the child in inpatient care, love and acceptance correlated most strongly with parental coping. PMID- 11896853 TI - Patient initiatives in psychiatric care concerning shame in the discussion in co operative team meetings. AB - The aim of this paper is to describe patients' experiences of shame and the way in which this experience is discussed in co-operative team meetings in acute psychiatric care. As an experience, shame is described as a painful and ugly feeling, which results in personal devaluation, isolation and a feeling of inferiority compared to others. This paper is based on 11 videotaped episodes of co-operative team meetings in two psychiatric units. The study approach was narrative. Shame was found to be the core narrative. The narrative of shame was laid out as an experience, and shame caused difficulties in the patients' daily lives and finally led to feelings of difference and loneliness. When the psychiatric patients and significant others tried to speak about the experience of shame with the professionals, the professionals either did not respond or changed the topic of discussion by asking questions about the patient's daily life and rationalizing the experience of shame. The participants did not share a common objective in the co-operative team meetings. Further research is needed to find new ways of co-operative team discussion and to develop the co-operative team meetings towards a patient-orientated model. PMID- 11896854 TI - Problems of young people in community psychiatric care. AB - The purpose of this study was to describe the problems of young people who need professional help and who have been referred to community psychiatric care. The aim was to develop both the type of care and the system of referral. The research data comprised all referrals (n = 184) to the outpatients' youth psychiatric clinic of one university hospital in 1997, and documentation related to the adolescents' interviews on admission. The data were analysed by both deductive and inductive content analysis. Young people's problems could be identified in all central spheres of life as difficulties in relation to the self, school, parents, peers, dating and the future. The difficulties were interrelated and varied in their degree of seriousness. When assessing young people's difficulties and need for care, focus should be made on the inner world of the young person, as well as on their behaviour within the different spheres of life. Only in this way is it possible to get a full picture of the adolescent's individual situation and need for help. PMID- 11896856 TI - Perspectives of collaboration/non-collaboration in a mental health inpatient setting. AB - Research investigating collaboration in health care has mainly taken place in general settings or has focused on the professional viewpoint. The initial aim of this study was an attempt to illuminate and explore 'collaboration' from the perspective of a person with a clinical diagnosis of schizophrenia using a hermeneutic-phenomenological approach. Through in-depth conversation, it became apparent that, rather than describing good examples of collaboration, the participant spontaneously drew attention to episodes of 'non-collaboration' and the difficulties he had experienced in joint decision-making in an inpatient mental health setting. Incorporated are extracts from a reflective journal to reveal the prejudices and preunderstandings of the researcher, who is a mental health nurse. The need for further research encompassing the views of those with a diagnosis of severe and enduring mental illness is underlined. PMID- 11896855 TI - Developing and testing methods for improving patient-oriented mental health care. AB - There is a trend in nursing policy and practice towards allowing patients to participate actively in their treatment and the services they use. The author concludes, based on an earlier qualitative study, that psychiatric patients in a hospital environment want a more active role in their treatment and that psychiatric nurses also aim at more active patient participation. In mental health care, professionals often define the needs of patients in terms of their own expertise and tend to overlook the variety of the service users' needs. The need to improve and empower psychiatric patients is a considerable challenge to present-day nursing. The purpose of this paper is to describe the theoretical background of tools used to assess patients' involvement in mental health care and the process by which credibility can be determined in rating the panel phase of instrument construction. The instruments, based on a qualitative study using grounded theory, produced a model of patient initiatives in psychiatric nursing. Content validity refers to the determination of the substantive representativeness or relevance of the items of an instrument. Face validity has been defined as validity conferred by a lay persons' acceptance that a procedure, statement or instrument appears to be sound or relevant. In the panel rating, the raters were experts. They evaluated the readability, consistency and content validity of the instruments' items. In the instrument for nurses, the content validity was 0.84 and consistency 0.91; this corresponds to 0.91 and 0.95, respectively, in the instrument for patients. PMID- 11896857 TI - Responding to a violent incident: physical restraint or anger management as therapeutic interventions. AB - By finding more feasible alternatives to managing aggression which can be effectively used in health care settings, as well as expanding nursing knowledge on the detrimental effects of restraints, forensic clinicians can use their creativity and resources to enhance the quality of life for their patients using beneficial therapeutic alternatives to restraint. A closer examination of alternatives to managing aggressive behaviour leads the author to the introduction of a case study, promoting non-touch interventions. The importance of this paper lies in reorienting forensic clinicians away from the perceived ideologies of physical restraint as a primary intervention, moving towards the productive use of anger management. PMID- 11896858 TI - Anorexia nervosa and culture. AB - Anorexia nervosa is currently considered a disorder confined to Western culture. Its recent identification in non-Western societies and different subcultures within the Western world has provoked a theory that Western cultural ideals of slimness and beauty have infiltrated these societies. The biomedical definition of anorexia nervosa emphasizes fat-phobia in the presentation of anorexia nervosa. However, evidence exists that suggests anorexia nevosa can exist without the Western fear of fatness and that this culturally biased view of anorexia nervosa may obscure health care professionals' understanding of a patient's own cultural reasons for self-starvation, and even hinder their recovery. PMID- 11896859 TI - Quality of treatment following police detention of mentally disordered offenders. AB - There is a growing interest in patients' perspectives in relation to the treatment and management of mentally disordered offenders at all levels of service delivery. This paper reports on a group of mentally ill individuals who were admitted to a place of safety following detention by the police. The principal purpose of the study was to determine the quality of care that the patient experienced under section 136 of the Mental Health Act (1983) from police detention to hospital admission. The major focus concerned the anticipated disparity between the non-professional police interactions and those of the mental health professional. The method employed was a grounded theory analysis of audio-taped interviews utilizing a three-level coding of data. The results indicated a general dissatisfaction with the quality of care and treatment from both police and professionals, but with the former being viewed as acceptable whilst the latter was considered unacceptable. The patients made a series of recommendations for detention in places of safety. PMID- 11896860 TI - Care systematization in psychiatric nursing within the psychiatric reform context. AB - The aim of this study was to approach care systematization in psychiatric nursing in two psychiatric disorder patients who attended 'Nossa Casa', Sao Lourenco do Sul, RS, Brazil. Nossa Casa services psychiatric patients in the community, focussing on: (i) permanence in their environment, allowing patients to remain close to their families and social spheres; (ii) integral attendance to meet individual needs; (iii) respecting individual differences; (iv) rehabilitation practices; and (v) social reinsertion. Concepts and assumptions of the psychiatric reform and the Irving's nursing process were used as theoretical methodological references to elaborate this systematization. A therapeutic project for the psychiatric patient was elaborated, in accordance with the interdisciplinary proposal accepted by Nossa Casa. Interdisciplinary team intervention, guided by a previously discussed common orientation and defined through an individualized therapeutic project, allowed for an effective process of psychosocial rehabilitation. The authors concluded that a therapeutic project based on the mentioned premises leads to consistent, comprehensive, dialectical and ethical assistance in mental health, thereby reinstating the citizenship of psychiatric patients. PMID- 11896861 TI - Othering and psychiatric nursing. AB - This paper is a theoretical exploration of the concept of Othering in relation to psychiatric nursing. The concept of Othering is examined by working the dualisms of same/other in relation to the dualisms of east/west, man/woman and reason/unreason. Othering is also examined in relation to the construction of knowledge and reality. The role of psychiatry and implications for nursing practice are discussed. Othering is found to be a complex problem which is an inevitability of nursing practice. Possible solutions to the problem of Othering are presented. PMID- 11896862 TI - An action research project in a night shelter for rough sleepers. AB - From October 1999 to June 2000, an action research project was undertaken in a homeless night shelter called Jimmy's. This project was grounded in user consultation and sought the involvement of staff and management to institute tangible improvements in service delivery using the Power Audit. A brief overview of Jimmy's is given, then this research is placed in local and national context by describing policy development in homelessness. A brief description is given of the research methodology and a short description of the Power Audit. Following this, the lives and experiences of the guests (Jimmy's residents) are conveyed using ethnography. This tells of the development of relationships with guests, staff and management without which the project could not have succeeded. Finally, an overview is provided of the interview content and the practical changes made. PMID- 11896863 TI - Legal, social, cultural and political developments in mental health care in the UK: the Liverpool black mental health service users' perspective. AB - Documentary evidence suggests that attitudes among local health and social services professionals towards the concept of user involvement in health and social care remain deeply polarized, a position characterized by commentators simultaneously as praise and damnation. Perhaps user involvement in health and social care will enhance, and it appears to resonate with the logic of, participatory democracy, in localities where the centralization of power has posed questions as to the nature and purpose of local governance in public services provision. The problems experienced by Britain's black and ethnic minorities within the mental health system have been the subject of exhaustive social inquiry. This essay attempts to explore the way in which legal, social, cultural, and political developments interface with mental health care practice in the UK, in order to assist those responsible for mental health services provision to deliver services that are in line with the Government's expectation of a modernized mental health service that is safe, sound, and supportive. An exploration of these developments within the European, national (UK), and local (Liverpool) contexts is undertaken. An appropriate local response to national priorities will ostensibly cut a swathe through the barriers confronted by the ethnic minority mental health service user in the cross-cultural context, an important prerequisite for the implementation of genuine user involvement. PMID- 11896864 TI - 'Partnership': a co-operative inquiry between Community Mental Health Nurses and their clients. 2. The nurse-client relationship. AB - This paper describes the output of a co-operative enquiry between Community Mental Health Nurses and their clients. Two nurses and two clients volunteered to participate as co-researchers and co-subjects with two facilitators in a co operative inquiry group. The subject of the inquiry, agreed by the group, was the relationship between the nurse and client. The description of the nurse-client relationship and the influences on it is extremely rich and potentially useful with implications for clinical and managerial practice. Despite the limitations of the study, co-operative inquiry appears to be a useful vehicle for contributing to an increased understanding of the nurse-client relationship with potential for further research and development. Unsurprisingly, when both the relationship characteristics and the health care context are appropriate there seems to be more positive outcomes for both nurse and client. Within the current health policy context there are increased external demands with the potential to disrupt this relationship. Nurses need to be aware of the potential effects of these so that the relationship and the client do not suffer and health service managers need to be aware of the contribution their behaviour may have on the nurse-client relationship. PMID- 11896865 TI - Comparative randomized study: administration of natural and synthetic surfactant to premature newborns with respiratory distress syndrome. AB - BACKGROUND: The respiratory distress syndrome (RDS) in premature newborns has been etiologically correlated to immature lungs and specifically with surfactant deficiency. Exogenous administration of surfactant is nowadays considered to be the treatment of choice. In this paper we attempt a comparison of clinical results from the administration of natural Alveofact and synthetic Exosurf surfactants in premature newborns with respiratory distress syndrome. METHODS: The study subjects were 92 premature newborns who had been hospitalized in the Department of Neonatology, of the University of Crete. A total of 42 subjects received synthetic surfactant and 50 subjects received natural surfactant. The surfactant was administered in one to three doses, depending on respiratory support requirements. RESULTS: The time of administration was a little longer for the natural surfactant group. The duration of mechanical ventilatory support, requiring oxygen, the duration of hospitalization and the percentage of increase of arterial alveolar partial pressure oxygen ratio (a/APO2) were slightly higher for the synthetic surfactant group. The mortality rate during the neonatal period (28th day) was higher for the synthetic surfactant group than for the natural surfactant group (38.1 vs 24%). A similar tendency was noticed also as regards to complications, e.g. pneumothorax (11.2 vs 5.2%; relative risk (RR) 0.27) intraventricular hemorrhage (34.6 vs 21.1%; RR 0.61), septicemia (11.5 vs 5.2%; RR 0.46) and bronchopulmonary dysplasia (12.5 vs 2.8%; RR 0.22). CONCLUSION: The use of natural surfactant seems to offer more advantages in comparison with its synthetic counterpart. PMID- 11896866 TI - Changes of the physiological parameters of very low-birthweight infants with chronic lung disease treated with dexamethasone. AB - BACKGROUND: Dexamethasone is widely used for the treatment of chronic lung disease (CLD), but its mechanism of action is still not clearly understood. METHODS: Respiratory status, bodyweight, blood pressure, urine volume, fluid intake and nutrient intake were investigated in 12 infants with CLD during treatment with dexamethasone. RESULTS: The mean gestational age of the patients was 26.3 +/- 2.5 weeks and their mean birthweight was 807 +/- 232 g. Treatment with dexamethasone was started at a mean age of 41 +/- 23 days. The ventilatory index (VI) improved after treatment was started. Blood pressure and urine volume increased significantly after treatment, but weight gain was poor during this time. Fluid and nutrient intake did not change before and after treatment. The degree of improvement in the VI after treatment was significantly correlated with an increase in urine volume. CONCLUSIONS: The results suggest that a rise in blood pressure as a result of dexamethasone treatment and the subsequent diuretic effect of this rise may play a role in improvement in respiratory status. PMID- 11896868 TI - Increased endothelin in infants of pre-eclamptic mothers. AB - OBJECTIVE: Endothelin (ET), a vasoactive mediator, like others which originate from the endothelium or circulating cells, may participate in myocardial injury. In full-term neonates of mild pre-eclamptic mothers (NMPM), it identifies minor myocardial damage missed by other biochemical markers. The present study was designed to determine the diagnostic value of ET concentrations in NMPM. METHODS: Seventeen NMPM were studied (10 boys and seven girls), and 17 healthy full-term (nine boys and eight girls) were selected for a control group. Birthweights in NMPM and control group were 2950 g (2.300-3.850) and 3.350 g (2.650-4.000), respectively. The ET measured by the radioimmunoassay method, Troponin T (TnT) were measured by enzyme-linked immunosorbent assay method and creatine kinase (CK MB) determined in chemical 1 analysis. RESULTS: Serum ET and TnT concentrations in NMPM (3.32 pg/mL vs 0.74 ng/mL) were significantly higher than the control group (0.82 pg/mL vs 0.10 ng/mL) (P < 0.01 and < 0.001). CONCLUSION: Our study demonstrated high levels of ET, cardiac TnT in NMPM, presumably associated with myocardial damage in NMPM. PMID- 11896867 TI - Preliminary study on DNA damage in non breast-fed infants. AB - BACKGROUND: There are many advantages of human milk for infants, including protection against cancer development and the advantages have been emphasized in several studies. In this study, infants fed by human milk has been compared with those fed by cow's milk concerning DNA damage. METHODS: The level of genetic damage in the peripheral blood lymphocytes of infants who were fed mainly by cow's milk and breast milk has been studied by sister chromatid exchange (SCE) analysis, which is a sensitive measurement of chromosomal damage. Each group consisted of 30 infants, whose ages ranged from 9 to 12 months. RESULTS: A significant increase (P < 0.0001) was found in the frequencies of SCE of infants not breast-fed (n = 30, mean SCE/cell +/- SD: 8.66 +/- 1.15) compared to those who were breast-fed (n = 30, mean SCE/cell +/- SD: 4.93 +/- 0.82). CONCLUSION: To our knowledge, there has been no published study investigating SCE ratio regarding DNA damage in infants not breast-fed. Molecular mechanism of DNA damage caused by the absence of human milk protection is a subject of future investigations. PMID- 11896869 TI - Longitudinal evaluation of anthracycline cardiotoxicity by signal-averaged electrocardiography in children with cancer. AB - BACKGROUND: The aim of the present study was to investigate the detection of anthracycline cardiotoxicity by signal-averaged electrocardiography (SAE) in children with cancer. METHODS: There were 29 patients with a cumulative anthracycline (ATC) dose of 75-600 mg/m2. None of them had congestive heart failure. Patients underwent SAE just before (detection of chronic cardiotoxicity) and just after (detection of acute cardiotoxicity) ATC therapy. Echocardiography and Holter electrocardiography were performed at the same time. The rates of abnormal SAE, echocardiography, and electrocardiogram findings were calculated and compared for every 100 mg/m2 of ATC. RESULTS: The SAE showed a significantly higher detection rate for acute cardiotoxicity was at a cumulative ATC dose of less than 400 mg/m2 when compared with other methods (P < 0.05). The lowest dose at which acute cardiotoxicity was detected by SAE was 117.3 mg/m2. The detection of chronic cardiotoxicity by SAE was significantly higher at a cumulative ATC dose of 100-400 mg/m2 when compared with other methods (P < 0.05), and the lowest value showing toxicity was 373.3 mg/m2. The lowest ATC dose causing chronic cardiotoxicity was significantly lower in patients less than 2-years-old (120.0 +/- 28.3 mg/m2) than in the other age groups (P < 0.05). CONCLUSIONS: Acute and chronic ATC cardiotoxicity were detected by SAE at lower cumulative doses compared with other methods. The technique of SAE was a potentially useful method for detection of cardiotoxicity among those investigated and it provides useful information on subclinical cardiac dysfunction in patients receiving ATC therapy. PMID- 11896870 TI - Retinal neurodevelopmental assessment with electroretinogram in patients with biliary atresia. AB - BACKGROUND: To evaluate the effect of n-3 polyunsaturated fatty acid (PUFA) deficiency on the development of retinal function in children with biliary atresia (BA), we examined serum fatty acid levels and performed electororetinogram (ERG) in patients with BA. METHODS: The study group was composed of one male and four female BA patients (8-14 years) with serum bilirubin levels ranging from 0.40 to 1.48 mg/dL. All of the subjects were born as full-term infants. The fatty acid composition of total lipids in serum was analyzed by gas chromatography before the Kasai operation, approximately 10 months after the Kasai operation, and at the time of the ERG study. The ERG was recorded using corneal contact lens electrodes. RESULTS: Two of the five patients showed decreased levels of arachidonic acid and docosahexaenoic acid (DHA) before and after the operation, but no abnormal findings on ERG were detected in these patients. The other three patients had decreased levels of alpha-linolenic acid or DHA after the operation, but again, no abnormalities were found on ERG. CONCLUSION: These results suggest that insufficiencies of DHA and other n-3 PUFA in full-term infants might not have an influence on later ERG results. PMID- 11896871 TI - Attenuated nitrergic inhibitory neurotransmission to interstitial cells of Cajal in the lower esophageal sphincter with esophageal achalasia in children. AB - BACKGROUND: Esophageal achalasia (EA) is a rare disease in children, the etiology and pathogenesis of which remain controversial. Previous studies have suggested that a specific class of interstitial cells of Cajal (ICC) act as mediators in nitrergic inhibitory neurotransmission in the lower esophageal sphincter (LES). The aim of this investigation is to clarify the status of ICC and nitrergic inhibitory neurons in the LES of EA using immunohistochemistry. METHODS: Specimens were obtained from two patients with EA (aged 6 and 10 years) and two patients with esophageal carcinoma (aged 56 and 63 years) not involving the lower esophagus as controls. Immunohistochemistry was used to study the distribution of ICC and nitrergic inhibitory neuron. RESULTS: The LES contains the c-kit positive ICC in the muscle layers, which form close relationships with nitric oxide synthase (NOS)-containing nerve fibers in the controls. The distribution of ICC was almost the same between samples with EA and controls. However, such nerve fibers were absent in EA with a longer duration of the symptoms, but were reduced in a shorter duration. CONCLUSIONS: Decreased nitrergic inhibitory neurotransmission to ICC in LES is a possible cause of sphincter achalasia in pediatric patients with EA. The decrease in NOS-positive neurons of patients with achalasia may be gradual, which may account for the long duration of symptoms prior to treatments. Further advancement of esophageal motility damage was suspected in pediatric EA. PMID- 11896872 TI - Bodyfat percentage in girls increased steadily with age and percentile rank of body mass index. AB - BACKGROUND: The purpose of the present study was to determine the body composition of elementary school children by bioelectrical impedance analysis (BIA) method with a subject in a standing position. The method is frequently used in Japan. The other aim was to evaluate the relationship between the body composition and percentile rank of the body mass index (BMI) in Japanese children. METHODS: The number of subjects were 1042 children (530 boys and 512 girls aged from 6- to 12-years-old) from an elementary school. The bioelectrical impedance (BI) in the standing position was measured late in the morning before lunch. The fat percentage was derived from the body density according to the formula of Brozek et al. Each percentile value of BMI for each age and sex was determined from the frequency table of height and weight published by the Ministry of Education in Japan. RESULTS: The fat percentage in both boys and girls was significantly correlated with the BMI, however, girls showed a closer linear relation than boys. The fat percentage in girls increased steadily with age and percent rank of the BMI. The fat percentage in boys was scattered in a wide range at each percentile rank of the BMI. CONCLUSIONS: The fat percentage measured by the BIA in the standing position is closely associated with the percentile rank of the BMI in elementary school girls. For boys, it will be necessary to compare data among different types of BI measurement methods. PMID- 11896874 TI - Evaluation of the need for routine preoperative latex allergy tests in children. AB - BACKGROUND: Recently, there was a great increase in allergic reactions to latex and this brought relatively more concern to the latex allergy. In this prospective study we aimed to identify the frequency of latex allergy in preoperative patients, and tried to clarify whether it is necessary to perform latex allergy tests routinely in the preoperative period or not. METHODS: A total of 188 children, aged 1-14, who were admitted for various operations, were randomly included in this study and of them, 181 completed the study. Latex specific history was taken from all patients. Latex skin prick tests, challenge tests with latex gloves, total IgE and latex specific IgE measurements were performed. RESULTS: Of 181 children, two (1.1%) had positive latex skin prick tests. Latex challenge tests were negative in all children. Latex specific IgE was positive in 12 children (6.6%) as class II or higher, but no patient had allergic reactions in operations. History of repeated operations was a risk factor for latex sensitization. The risk was higher in the presence of both history of repeated operations and history of allergic disease. However, the risk was not higher in patients with the history of only allergic disease, compared to ones who had a history of neither repeated operations nor allergic disease. CONCLUSION: We conclude that routine preoperative latex allergy tests seem to be not necessary because of no allergic reactions during operation in spite of the sensitization of 6.6% detected by latex specific IgE. However, this should be investigated in larger studies. PMID- 11896873 TI - Psychosomatic disorders in children: an emerging challenge to health care in Japan. AB - BACKGROUND: Japan has been witnessing a considerable rise in the number of children presenting with psychosomatic disorders. Thus, there is a mounting demand for pediatricians to be trained in the caring of children with psychosomatic disorders. METHOD: A questionnaire based-survey was conducted to investigate the average number of working hours for the first year (Postgraduate Year 1 (PGY1)) pediatric residents at Keio University Hospital. The same survey was conducted retrospectively with doctors with at least 10 years of experience in child health care. Another survey was conducted at our 27 affiliated hospitals with respect to the number of outpatients and patients presenting with psychosomatic disorders. RESULTS: The survey showed that, in the ward for preschool and school-age children, PGY1 residents spent on average 78.9 h per week. Time required for care of psychosomatic disorders was more than 30% of the total amount of time allotted for information acquisition, interviews and consultations, as well as conferences and discussions. There was virtually no time spent on the care for psychosomatic disorders in our pediatric residency program 10 years ago. It is an implication of the survey that the apparent number of patients with psychosomatic problems is not dependent on the size or location of the hospital. CONCLUSION: General pediatricians will be spending more time than ever before in the caring of children with psychosomatic disorders. Moreover, pediatricians trained for child health care for psychosomatic disorders are short of supply or non-existent, even in the major affiliated hospitals. PMID- 11896875 TI - Intracavernous application of diazepam: an alternative route of the seizure treatment--an experimental study in rabbits. AB - BACKGROUND: There is a general need to terminate seizures as soon as possible using anticonvulsant drugs via an intravenous (i.v.) route, but it is often difficult to achieve a secure i.v. line during the seizure, especially in children. However, it has been demonstrated that high volumes of fluid can be injected into the corpora cavernosa. The purpose of this study was to evaluate the absorption properties of diazepam (DZ) after intracavernous (i.c.) administration and whether therapeutically significant plasma concentrations can be obtained or not. METHODS: Diazepam was administered to rabbits using both the i.v. and i.c route with a dose of 1 mg/kg. Blood samples were collected from the saphenous vein for a time period of 30 min. The levels of DZ in the blood were analyzed by high performance liquid chromatography and their blood profiles were obtained and compared. RESULTS: The insertions of the needle using this method were successful in all cases. The average time of inserting the needle was less than 5 s. There was no statistically significant difference between the i.v. and i.c. administration with regard of the blood DZ levels. Within 48 h after the experiment, none of the animals demonstrated any evidence of infection or disability. CONCLUSION: Our results demonstrated that commercially available parenteral DZ can be absorbed rapidly by the i.c. route in rabbits. Further studies are needed on the feasibility of this method before it is evaluated in humans. PMID- 11896876 TI - Decreased frequency of seizures in infantile spasms associated with lissencephaly by human herpes virus 7 infection. PMID- 11896877 TI - Localization-related epilepsy mimicking epilepsy with myoclonic absence in a patient with pachygyria. PMID- 11896879 TI - Two cases of diabetes mellitus type 1 accompanied by hallucinatory-delusional state. PMID- 11896878 TI - Severe heart failure due to ductal constriction caused by maternal indomethacin. PMID- 11896880 TI - Case of incidentally diagnosed vitamin D deficiency rickets: a review of literature from Japan and a proposal for reintroduction of vitamin D2. PMID- 11896881 TI - Detection of abnormal union of pancreaticobiliary junction by magnetic resonance cholangiopancreatography in a girl with acute pancreatitis. PMID- 11896882 TI - Tuberculous osteomyelitis of the sternum in an infant: a case report and review of the literature. PMID- 11896883 TI - Case report of fetus-in-fetu diagnosed in a neonate with trisomy 21. PMID- 11896884 TI - Atlanto-axial subluxation (Grisel's syndrome) associated with mumps. PMID- 11896885 TI - Foreword--Mizoribine: a new immunosuppressive drug for kidney diseases and rheumatic diseases. PMID- 11896886 TI - Mizoribine: mode of action and effects in clinical use. AB - Mizoribine is a new immunosuppressive drug and was authorized by the Japanese Government in 1984. The strong immunosuppressive activity of mizoribine was already demonstrated in various animal models, in renal transplantation and in steroid-resistant nephrotic syndrome. Recently, the remarkable clinical advantages of an imidazole for adults with rheumatoid arthritis, lupus nephritis and other rheumatic diseases were reported. Mizoribine is an imidazole nucleoside and the metabolites, MZ-5-P, exerts its activity through selective inhibition of inosine monophosphate synthetase and guanosine monophosphate synthetase, resulting in the complete inhibition of guanine nucleotide synthesis without incorporation into nucleotides. Thus, mizoribine is superior to azathioprine, in that it may not cause damages to normal cells and normal nucleic acid. PMID- 11896887 TI - Mizoribine as an effective combined maintenance therapy with prednisolone in child-onset systemic lupus erythematosus. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is one of the major collagen diseases in childhood. However, the pathogenesis of this disease still remains unknown. The disease is known as a chronic inflammatory disease. Since oral and intravenous corticosteroid therapy has been introduced into the treatment of SLE, the prognosis of patients has improved significantly. However, it has now become clear that there are limitations in the effectiveness, as well as adverse reactions when corticosteroids therapy is administered for a long-term period. Therefore, we have been attempting to improve the maintenance therapy of child onset SLE. METHODS: We have proposed and tested a new type of combination therapy using prednisolone (PSL) and mizoribine (MZR) in pediatric patients with SLE for maintenance therapy after the induction of remission. RESULTS: Our results showed that this combination therapy is more effective than the previous regimen. In addition, no significant side-effects were observed in our study. CONCLUSION: This combination therapy is still not perfect. Efforts should be continued to establish an optimal therapy regimen for child-onset SLE. PMID- 11896888 TI - Mizoribine in the treatment of rheumatoid arthritis and juvenile idiopathic arthritis. AB - BACKGROUND: Mizoribine (MZR), isolated from culture medium of the mold, is a novel immunosuppressant developed in Japan. It has been used in patients with renal transplantation, lupus nephritis, nephrotic syndrome and rheumatoid arthritis (RA). OBJECTIVES: To review MZR in regards to mechanism of action, pharmacokinetics, efficacy and safety in the treatment of rheumatoid RA and juvenile idiopathic arthritis (JIA). RESULTS: The drug MZR inhibits both humoral and cellular immunity in RA patients. It is completely excreted in the urine within 24 h, which contributes to the safety of MZR. A series of multicenter studies indicated that MZR was effective and safe in the treatment of RA. In JIA, however, there are only a few case reports reporting its efficacy and safety. CONCLUSION: A double-blinded multicenter study is needed to establish the efficacy, safety and indication of MZR in the treatment of JIA. PMID- 11896889 TI - Nephrotic syndrome and mizoribine in children. AB - BACKGROUND: The treatment of nephrotic syndrome (NS) has recently made dramatic progress. The ultimate purpose of treatment is that patients can lead a normal, disease-free life with no adverse effects from treatment. However, clear treatment guidelines remain to be established in the children with frequent relapsing NS (FRNS) and steroid-resistant NS (SRNS). The frequent use or large dose of steroids may lead to a serious adverse effect. Immunosuppressive drugs including cyclophosphamide and cyclosporin have been used to induce lasting remission, thereby sparing patients from further exposure to steroids. However, these drugs have both acute and chronic side-effects. Mizoribine is a relatively safe and effective immunosuppressant. The report discusses the role of mizoribine in FRNS and SRNS in children. METHODS: The papers in experimental and clinical studies about mizoribine in NS were reviewed. Our experiences of mizoribine were also added. RESULTS: Mizoribine has been reported as an effective and safe drug for patients with FRNS. However, the efficacy of mizoribine has differed among various reports, depending on the dose given. There have yet to be any conclusive reports on the effects of mizoribine in SRNS in children. CONCLUSIONS: Better results might be obtained if the doses of mizoribine increased in children with FRNS. When 5 mg/kg was used, no serious adverse effects were seen, therefore this dose may be safe and effective. An investigation of appropriate, effective and safe doses of mizoribine should be examined in the future. In patients with SRNS, large doses of mizoribine of more than 5 mg/kg might be effective, while the combined use of mizoribine and cyclosporin or methylprednisolone pulse therapy might also be even more effective. Further studies are called for to examine the use of large doses of mizoribine. PMID- 11896890 TI - Mizoribine treatment for childhood IgA nephropathy. AB - BACKGROUND: There is currently no established therapy for childhood IgA nephropathy (IgAN). Mizoribine, a newly developed immunosuppressive agent characterized as a safe and well-tolerated drug, has been widely used in diverse conditions. Our preliminary study demonstrated that mizoribine could reduce the amount of proteinuria in children with IgAN. The present study was conducted to confirm this finding. METHODS: Ten children with IgAN (median age 13.5 years) of moderate histological severity were enrolled. None of them had been previously treated by immunosuppressants. Mizoribine was administered orally for a median period of 20.5 months. We compared the urinary protein excretion expressed as the ratio of urinary protein to urinary creatinine (UP/UC) and the hematuria evaluated as the level of occult blood by dip-stick (OB score). Renal histology was also examined in three patients using paired biopsy specimens obtained both before and after treatment. We performed blood examinations regularly to monitor the toxicity and plasma concentration of mizoribine. RESULTS: The median observation period was 44.5 months, consisting of a median 13.0 months before therapy, 20.5 months during therapy and 12.0 months after therapy. Significant reductions in both UP/UC and OB score were induced by mizoribine (P < 0.05). Renal mesangial proliferation was also improved. Plasma peak levels of mizoribine varied from 0.30 microg/mL to 1.23 microg/mL and were not associated with its effectiveness. No adverse effects were observed during the therapy, although a slight decrease in leukocyte count was noted. CONCLUSION: Mizoribine can be an alternative drug for childhood IgAN with moderate severity because it results in a significant reduction of proteinuria and hematuria with histological improvement and causes far fewer complications compared to the conventional immunosuppressants. PMID- 11896891 TI - Role of mizoribine in renal transplantation. AB - Renal transplantation is the optimal form of therapy for children and adolescents with end-stage renal disease. Usually histocompatibility differences exist between donor and recipient, so it is necessary to modify or suppress the immune response to enable the recipient to accept a graft. Calcineurin inhibitors (CNI), which include cyclosporin (CsA) and tacrolimus (FK506), give many benefits on the outcome after renal transplantation, but have some toxic effects, especially nephrotoxicity. Therefore, inhibitors of purine synthesis revived as newer generation of more specific inhibitors, mizoribine (MZ) and mycophenolate mofetil (MMF). The Japanese pediatric renal transplantation clinical study group attempted to reduce and then discontinue steroid administration in combination with another three immunosuppressive drugs, CsA, MZ and anti-lymphocyte globulin (ALG). This study showed good clinical results. Mizoribine is an effective immunosuppressive drug in human renal transplantation. However, it is not as popular as other inhibitors of purine synthesis, such as azathioprine (AZA) and MMF, because MZ has been used mainly in Japan and infrequently in other countries. However, MZ is a more useful immunosuppressive drug than AZA, when it is used in combination with CNI. PMID- 11896893 TI - Volume 18 Acknowledgments. PMID- 11896892 TI - Freedom to Be. PMID- 11896894 TI - The global initiative for chronic obstructive lung disease: Gold standards and the Asia-Pacific perspective. PMID- 11896895 TI - Pulmonary hypertension in patients with chronic obstructive pulmonary disease: recent advances in pathophysiology and management. AB - In patients with chronic obstructive pulmonary disease (COPD), pulmonary hypertension (PH) is associated with a worse prognosis. Recently, information has been increasing concerning the cellular and molecular aspects of the pathophysiology of PH in COPD. The most striking finding is the role of vascular endothelial cells and endogenous mediators released by these cells. Endothelial cell-dependent relaxation is impaired in COPD patients with PH. Moreover, vascular remodelling in these patients is mainly responsible for irreversible PH in advanced COPD. Smoking cessation will slow down the progression of the disease process and may prevent the development of PH in COPD. The timing of initiation of long-term oxygen therapy is important for the effective management of PH in COPD. Research on therapeutic agents for the effective treatment of PH is still needed in the management aspect of patients with COPD. This review focuses on the recent advances in our understanding of the pathophysiology and treatment of PH in COPD. PMID- 11896896 TI - Increased circulating levels of soluble Fas ligand are correlated with disease activity in patients with fibrosing lung diseases. AB - OBJECTIVE: The Fas-Fas ligand (FasL) pathway is one of the important apoptosis signalling molecule systems. We previously determined that this pathway may be involved in the pathogenesis of fibrosing lung diseases. In the present study, we evaluated the clinical significance of the levels of soluble forms of Fas (sFas) and FasL (sFasL) in serum from patients with fibrosing lung diseases. METHODOLOGY: We measured sFas, sFasL, KL-6 (a measure of alveolar type II cell damage), surfactant protein D (SP-D), and surfactant protein A (SP-A) levels in serum from 35 patients with idiopathic pulmonary fibrosis (IPF), 17 patients with interstitial pneumonia associated with collagen vascular diseases (CVD-IP), and 13 normal healthy controls using enzyme-linked immunosorbent assays (ELISA). RESULTS: The serum levels of sFasL were significantly increased in patients with active IPF and CVD-IP, compared with those with inactive disease and controls. There was no significant difference in sFasL levels between patients with inactive disease and controls. Serum sFasL levels were significantly correlated with lactate dehydrogenase and KL-6 levels in IPF. The decrease in sFasL levels following corticosteroid therapy was not correlated with the clinical course of IPF. There was no significant difference in serum sFas levels between IPF or CVD IP patients and controls. CONCLUSIONS: Although further studies need to be performed on a large number of patients with histologically proven IPF or CVD-IP, it would seem that serum sFasL levels may reflect the activity of IPF and CVD-IP. PMID- 11896897 TI - Interleukin-1beta and tumour necrosis factor-alpha increase microvascular leakage in the guinea pig trachea. AB - OBJECTIVE: Airway microvascular leakage is considered to be an important component of airway inflammation in asthma. In the present study we examined the effect of interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNFalpha) on airway microvascular leakage in vivo. METHODOLOGY: Tracheal Evans blue extravasation was examined in an isolated tracheal segment, in anaesthetized mechanically ventilated guinea pigs. Baseline tracheal microvascular leakage was measured in five animals. As a control group for aerosol challenge, the isolated tracheal segment (n = 5) underwent saline aerosol challenge. To test whether a combination of IL-1beta (10 ng/mL) and TNFalpha (100 ng/mL) induced Evans blue extravasation, the trachea was exposed to an aerosol of these cytokines (n = 5). As a positive control the tracheal segment was challenged with histamine aerosol (5 x 10(-2) mol) (n = 3). All aerosol challenges were for 1 min. RESULTS: TNFalpha and IL-1beta aerosol challenge significantly increased Evans blue extravasation (28.9 +/- 1.6 microg/g wet tissue, mean +/- SE) compared to saline challenge (13.8 +/- 3.0 microg/g; P < 0.05). Tracheal dye extravasation without aerosol challenge, was not significantly different from saline-challenged animals (17.5 +/- 2.9 and 13.8 +/- 3.0 microg/g, respectively). Histamine significantly increased Evans blue extravasation (50.1 +/- 4.8 microg/g; P < 0.05) compared to saline challenge. CONCLUSION: Pro-inflammatory cytokines, TNFalpha and IL-1beta are able to induce significant microvascular leakage in the guinea pig trachea. PMID- 11896898 TI - Adenovirus vector-mediated transfer of 9 kDa granulysin induces DNA fragmentation in HuD antigen-expressing small cell lung cancer murine model cells. AB - OBJECTIVE: Granulysin is a tumoricidal molecule secreted by cytotoxic T cells (CTL) and natural killer (NK) cells, that induces apoptotic cell death in tumour cells. It has been demonstrated that small cell lung cancer (SCLC) cell lines are susceptible to NK cells and lymphokine activated killer (LAK) cells, and HuD antigen is assumed to be a target molecule on SCLC cells for host cellular immunity. METHODOLOGY: In order to understand the mechanism of sensitivity of SCLC to cellular immunity, we evaluated granulysin-induced apoptosis using mouse adenocarcinoma Colon 26 (Colon 26/HuD) cells transfected with the 9 kDa active form of granulysin using an adenovirus vector as a murine model of SCLC cells. RESULTS: Adenovirus vector-mediated transfer of 9 kDa granulysin increased DNA fragmentation in Colon 26/HuD cells 2.5-fold and suppressed Colon 26/HuD proliferation by 21% on day 3 (P < 0.05 for each value) compared with the control adenovirus vector transfer. In contrast, adenovirus vector-mediated transfer of 9 kDa granulysin did not increase DNA fragmentation nor suppress the proliferation of Colon 26 parent cells. CONCLUSIONS: The sensitivity of HuD-expressing tumour cells to granulysin is likely to partially explain the susceptibility of SCLC to cell-mediated immunity. PMID- 11896899 TI - Budesonide reduces sensitivity and reactivity to inhaled mannitol in asthmatic subjects. AB - OBJECTIVE: The aim of the study was to investigate whether treatment using inhaled corticosteroids decreases airway responsiveness to inhaled mannitol in asthmatic subjects. METHODOLOGY: Before treatment or a change in treatment with inhaled corticosteroids, 18 asthmatic subjects had measurements of lung function and airway sensitivity to mannitol taken and they completed a self-administered questionnaire on asthma symptoms. The procedure was repeated 6-9 weeks after taking 800-2400 microg/day of budesonide. RESULTS: There were significant reductions in airway sensitivity (provoking dose to induce a 15% fall in FEV1 (PD15)) and airway reactivity measured by the response dose ratio (RDR; final percentage fall FEV1/total dose of mannitol administered). The PD15 (Gmean (95%CI)) increased from 78 mg (51, 117) before treatment to 289 mg (202, 414) following treatment (P < 0.001). All subjects had a significant increase beyond the repeatability of 0.9 doubling doses with seven subjects becoming unresponsive. There was a 4.2 (3.4, 4.9)-fold improvement in the RDR with the value before the treatment period 0.18 (0.12, 0.28) decreasing to 0.04 (0.03, 0.08) following treatment (P < 0.001). These improvements were associated with significant improvements in lung function and symptom severity. CONCLUSION: Treatment with the inhaled corticosteroid budesonide caused a decrease in airway sensitivity and reactivity to inhaled mannitol and this was associated with expected improvements in lung function and symptoms. PMID- 11896900 TI - Pulmonary diffusion capacity in patients with systemic lupus erythematosus. AB - OBJECTIVE: The aim of this study was to clarify the characteristics of pulmonary function tests (PFT), especially carbon monoxide diffusion capacity (DLCO), and their correlation with clinical features and immunological findings in patients with systemic lupus erythematosus (SLE). METHODOLOGY: Vital capacity (VC) and DLCO were analysed retrospectively in 110 sequential Japanese SLE patients with active disease between 1985 and 1999. In 38 patients, serial measurements of PFT were also assessed during high-dose corticosteroid therapy. RESULTS: DLCO was reduced in 52 patients (47%) and a restrictive impairment of PFT was observed in nine patients (8%). The prevalence of pulmonary fibrosis was 13%. Reduced DLCO was frequently observed, even in patients with neither pulmonary fibrosis nor a restrictive pattern. No correlation between immunological data and reduced DLCO was found, except for the presence of anti-RNP. Patients with Raynaud's phenomenon showed a higher prevalence of DLCO impairment than those without this phenomenon. Although immunological parameters improved significantly after the corticosteroid therapy, no significant change in the level of DLCO was observed. CONCLUSIONS: Impairment of DLCO was frequently observed in patients with SLE who had no clinical respiratory abnormalities. DLCO impairments were correlated with Raynaud's phenomenon clinically, and the presence of anti-RNP immunologically. No significant correlation was found between impairment of DLCO and disease activity of SLE. PMID- 11896901 TI - Decline in FEV1 in patients with PiZ alpha-1-antitrypsin deficiency: the Australian experience. AB - OBJECTIVE: Alpha-1-antitrypsin (alpha1antitrypsin) deficiency is a rare hereditary disorder which characteristically presents with emphysema at an early age. The aim of the present study was to determine whether the rate of decline of lung function in alpha1antitrypsin-deficient subjects in Australia was similar to that found elsewhere. METHODOLOGY: Patients registered with the Australian Alpha 1-Antitrypsin Replacement Program were studied. All patients (n = 50) had a serum alpha1antitrypsin concentration of < 0.3 g/L and had had spirometry measured over at least 2 years. They were compared with a group of normal volunteers (hospital staff, n = 107) with normal alpha1antitrypsin levels and phenotypes and with no clinical history of lung disease. All had spirometry measured for periods ranging from 2 to 6 years. The rate of decline of forced expiratory volume in 1 s (FEV1) for each subject was calculated by least squares linear regression using FEV1 against the time from entry into the study. RESULTS: The group mean (+/- SD) rate of decline in FEV1 was significantly greater (P < 0.01) in the alpha1antitrypsin deficient patients (88 +/- 71 mL/year) than for the normal controls (-15 +/- 48 mL/year). There was no difference in decline in FEV1 when the data was analysed for gender and for index versus non-index cases. CONCLUSION: The results confirm previous reports of an accelerated rate of decline of FEV1 in patients with alpha1antitrypsin deficiency. Our results indicate that the rate of decline of lung function in alpha1antitrypsin deficient subjects in Australia is similar to that found in reported series from elsewhere. PMID- 11896902 TI - Extrathoracic staging of non-small cell bronchogenic carcinoma: relationship of the clinical evaluation to organ scans. AB - OBJECTIVE: The aim of this study was to investigate the value of systemic evaluation of extrathoracic extension of non-small cell lung cancer and to assess the value of the clinical evaluation in detecting extrathoracic metastases. METHODOLOGY: The study included 90 patients [87 men, three women; mean age 57.5 years (range 28-76)] with potentially resectable non-small cell carcinoma. Fifty two were squamous cell carcinomas and 38 were adenocarcinomas. Organ-specific and non-organ-specific clinical findings suggesting metastases were analysed and computed tomographic scans of the brain and abdomen and whole-body bone scanning were performed in all patients. RESULTS: Extrathoracic metastases were detected in 23 (25.5%) of 90 patients. The metastases were located in the following areas: brain (n = 12, 13.3%); bone (n = 9, 10%); liver (n = 5, 5.6%); and adrenal gland (n = 5, 5.5%). Histological analysis revealed metastases in 21.1% (11/52) of the squamous cell carcinomas and 31.6% (12/38) of the adenocarcinomas (P > 0.05). Eleven (47.8%) of the 23 patients with extrathoracic metastases had no organ specific clinical findings suggesting metastases. Eight patients with squamous cell carcinomas were intrathoracic T1N0 stage and in two (25%) of these patients extrathoracic metastases were detected. These patients had no organ-specific or non-organ-specific clinical factors suggesting metastases. CONCLUSIONS: Evaluation of extrathoracic extension should be routinely performed in all patients with newly diagnosed lung cancer. This approach will prevent many unnecessary thoracotomies. PMID- 11896903 TI - Clinical, laboratory findings and microbiologic characterization of bronchiectasis in Thai patients. AB - OBJECTIVE: The aim of the present study was to characterize the clinical features, underlying disease states, laboratory findings and microbiological characterization of bronchiectasis in Thai patients. METHODOLOGY: For a 2-year period all consecutive patients diagnosed with bronchiectasis at Phramongkutklao Hospital, Bangkok, Thailand, were recruited. Data including history, physical examination, underlying disease and laboratory studies were carefully reviewed and recorded. RESULTS: Fifty patients diagnosed with bronchiectasis were enrolled. Their mean age was 58 years. The most common background aetiology was tuberculosis. Six per cent of the patients were diagnosed as having diffuse panbronchiolitis. Normal chest radiographs were found in 10%. The common organisms isolated were Pseudomonas aeruginosa (20%), Haemophilus influenzae (14%), Klebsiella pneumoniae (14%) and Streptococcus pneumoniae (6%). Non tuberculous mycobacteria which included Mycobacterium kansasii and Mycobacterium chelonae were found in 6%. CONCLUSION: We report the characteristics of bronchiectasis in Thai patients. The most common identifiable aetiology was tuberculosis. PMID- 11896904 TI - Sample survey of drug-resistant tuberculosis in Henan, China, 1996. AB - BACKGROUND: There is little reliable data on the global drug resistance to tuberculosis (TB) as most of the existing data is based upon biased samples, is not standardized or was obtained using poor techniques. For this reason, the World Health Organization (WHO) and the International Union Against Tuberculosis and Lung Disease (IUATLD) developed a global project on anti-TB drug resistance surveillance (DRS) in 1994. China joined this project in 1995 and the province of Henan was selected as the first site for collection of representative samples to survey the prevalence of drug-resistant TB. METHODOLOGY: Standard drug susceptibility testing by the proportion method against streptomycin (S), isoniazid (H), rifampicin (R), and ethambutol (E) was performed with Mycobacterium tuberculosis isolated from 916 new cases and 456 previously treated cases. Treatment outcome of these patients has been evaluated according to the regimens and drug susceptibility patterns. RESULTS: Drug resistance among new cases to any drug was found to be 43.0% and any resistance: S, 32.5%; H, 31.0%; R, 20.7%; and E, 10.3%. Drug resistance among previously treated cases to any drug was 68.2% and any resistance: S, 52.2%; H, 49.3%; R, 48.3%; and E, 20.4%. The cure rate for new cases was 43.3% and 29.4% for previously treated cases. The poor cure rate resulted mainly from a high defaulter rate. CONCLUSION: Drug resistant TB was found to be highly prevalent in Henan and the cure rate remained poor. The results strongly indicated that Henan should take immediate action to improve the cure rate of patients through expansion of the introduction of the directly observed treatment short-course strategy. PMID- 11896905 TI - Regression of bilateral bronchiectasis with inhaled steroid therapy. AB - Bronchiectasis is defined as pathological and permanent dilatation of the bronchial tree. Affected patients suffer from chronic sputum production and usually slowly progressive airway destruction as a result of continued airway infection and inflammation. Regression of bilateral bronchiectasis has never been reported in the English literature. We report the case of a 60-year-old woman with longstanding progressive idiopathic bilateral bronchiectasis whose respiratory symptoms, including sputum, rapidly disappeared after commencement of inhaled budesonide. Repeat computed tomography assessment 40 months after commencement of inhaled steroid therapy, showed partial regression of bronchial dilation and resolution of small airways sepsis. In the absence of other possible explanations for the partial resolution of the bronchiectasis, the present case suggests a possible benefit of inhaled steroid therapy in bronchiectasis. PMID- 11896906 TI - A case of extrapleural empyema. AB - A 49-year-old man with diabetes mellitus and alcoholic liver cirrhosis presented with dyspnoea and fever. A chest computed tomography scan revealed three areas of loculated pleural effusion. Initially, the patient was thought to have an intrapleural empyema and was treated with intravenous antibiotics and closed drainage. However, as he did not improve, he was then treated with open drainage. During open drainage, the patient was diagnosed to have an extrapleural empyema and improved following open drainage treatment. PMID- 11896927 TI - Some hints concerning the shape of T-cell receptor structures. AB - Several models are proposed for T-cell antigen receptor (TCR) assembly and structure. However, there is little experimental data favouring directly either one or the other(s). The minimal complex appears to be composed of a TCRalphabeta/CD3deltaepsilon,gammaepsilon/zeta2 structure but at the cell membrane, multimers of this minimal structure may be formed. Quantitative cytofluometry has suggested three CD3epsilon chains for two TCRbeta (or TCRdelta) chains/complex. Such data should be repeated with monoclonal antibodies (MoAb) against extracellular (EC) parts of CD3delta or CD3gamma chains. In the present review, we have compared the TCR/CD3 assembly of pre-TCR, TCRgammadelta and TCRalphabeta containing complexes, and analysed the reactivity of antibodies (Abs) against the EC part of CD3delta chains. Our data suggest an alternative assembly pathway and structure of TCR/CD3 complexes. PMID- 11896928 TI - A pTalpha-negative subpopulation of CD25+ TN thymocytes revealed by a transgenic marker. AB - We have recently generated 5'lambda5-huTAC mice, which express the human CD25 (huTAC) gene under the control of the 5'-flanking region of the mouse lambda5 gene. The huTAC-transgene was expressed in pre-B cells but neither in mature B cells nor in T cells of these mice. In this report we demonstrate that the transgene is also transiently expressed by adult CD25+ CD3-CD4-CD8- (triple negative, TN) thymocytes and in fetal thymocytes. The huTAC+, in contrast to the huTAC- subpopulation of the CD44+CD25+ TN cells, was unexpectedly found not to express the pTalpha-gene. Still the huTAC+CD44+CD25+ TN cells reconstituted the development of both alphabeta and gammadelta lineage cells equally efficiently as the pTalpha-expressing huTAC- fraction, demonstrating that this pTalpha-negative subpopulation contained precursors for both T-cell lineages. Single cell reverse transcription-polymerase chain reaction (RT-PCR) experiments demonstrated that also in normal mice only a fraction of CD44+CD25+ and CD44-CD25+ TN cells expressed this gene. Taken together, these data indicate that huTAC transgene expression revealed a truly pTalpha-negative fraction of the CD44+CD25+ TN cells. The observation that not all precursors in the CD25+ TN population express the pTalpha-gene has important implications for the understanding of early T-cell development and T-cell lineage commitment. PMID- 11896929 TI - The role of an exposed loop in the alpha2 domain in the mouse MHC class I H-2Dd molecule for recognition by the monoclonal antibody 34-5-8S and the NK-cell receptor Ly49A. AB - Natural killer (NK) cells express major histocompatibility complex (MHC) class I specific inhibitory receptors. The region mediating the protective effect of the MHC class I molecule H-2Dd (Dd), recognized by the inhibitory receptor Ly49A, has been mapped to the alpha1/alpha2 domains. Here we have focused on an exposed loop in the N-terminal part of the alpha2 domain, which constitutes a major structural motif that differs between Dd (strong binding to Ly49A) and Db (weak binding to Ly49A at best). We mutated the residues 103, 104 and 107 in Dd to the corresponding amino acids in Db. The Dd mutant molecule retained the ability to be stabilized by a Dd-binding peptide. However, the mutation totally abolished the recognition by the conformational dependent monoclonal antibody (MoAb) 34-5 8S, known to inhibit the interaction between Dd and Ly49A. In addition, there was a marked impairment of the binding to Ly49A as evaluated by the ability of tetramers of the Dd mutant molecule to bind to Ly49A-transfected reporter cells and spleen cells. These results demonstrate that the introduced changes at positions 103, 104 and 107 directly or indirectly affect the epitopes for the MoAb 34-5-8S and the Ly49A receptor. PMID- 11896931 TI - Human cytomegalovirus (HCMV)-infected endothelial cells and macrophages are less susceptible to natural killer lysis independent of the downregulation of classical HLA class I molecules or expression of the HCMV class I homologue, UL18. AB - A number of reports have suggested that human cytomegalovirus (HCMV)-infected fibroblasts are resistant to natural killer (NK) lysis, and that the HCMV-encoded human leucocyte antigen (HLA) class I homologue UL18 may be responsible for this effect. While fibroblasts are easy to infect in vitro, their role in HCMV pathogenesis in vivo is unclear. Here, we have established systems to address NK recognition of infected endothelial cells and macrophages, two important HCMV cellular reservoirs in vivo. The HCMV-infected endothelial cells exhibited increased resistance to NK killing, and, in most experiments, infected macrophages demonstrated a decreased susceptibility to NK lysis. Infection with the mutant HCMV strain RV670, lacking the genes US1-9 and US11 that are responsible for downregulation of HLA class I molecules, also led to decreased NK susceptibility. Furthermore, reduced NK susceptibility was independent of the expression of the HLA class I homologue UL18, since cells infected with the UL18Delta HCMV strain were also less susceptible to NK killing. These results suggest that HCMV-induced resistance to NK cytotoxicity in endothelial cells and macrophages is independent of known pathways that interfere with the expression of cellular HLA class I A, B and C surface antigens and the HCMV encoded class I homologue UL18. PMID- 11896930 TI - Extensive CDR3H length heterogeneity exists in bovine foetal VDJ rearrangements. AB - Analysis of seven variable-diversity-joining (VDJ) gene rearrangements in B splenocytes from a 125-day-old bovine foetus revealed an extensive heavy-chain complementarity-determining region 3 (CDR3H) length variation (9-56 codons). Indeed, the global CDR3H size spectratyping of foetal VDJ rearrangements substantiated such an extensive heterogeneity and was comparable with that noted in peripheral B lymphocytes of adult cattle. These observations are in contrast to species such as humans with extensive germline combinatorial capability where shorter CDR3H length is noted early during B-cell development. Exceptionally long CDR3H (as in adult cattle) was noted in two foetal VDJ rearrangements encoded by a single germline VH gene. Further, two VH genes (gl.110.20 and BF2B5) were preferentially expressed in the foetal VDJ rearrangements. The DH gene-encoded CDR3H region of foetal VDJ rearrangements is remarkable for repetitive GGT (glycine) and TAT (tyrosine) codons that favour the recruitment of somatic hypermutations. It appears that closely related germline DH genes, preferentially used in the hydrophilic reading frame, encode varying CDR3H lengths early during B-cell ontogeny in cattle. A comparison of germline and expressed VH genes, especially in the CDR1 and CDR2, confirms that somatic hypermutations contribute to immunoglobulin (Ig)M antibody diversification in cattle. The biased nucleotide base use and high occurrence of 'hot-spot' triplet (AGPy; AG pyrimidine base) in the CDRs predisposes to somatic hypermutations. Overall, these observations suggest that extensive CDR3H length heterogeneity, including the generation of exceptionally long CDR3H (up to 56 amino acids), and somatic hypermutations contribute to IgM antibody diversification in cattle. The extensive CDR3H length heterogeneity early during the B-cell development may compensate for constraints imposed on antibody diversification owing to the limited germline sequence diversity of genetic elements in cattle. PMID- 11896932 TI - Identification of two Ikaros-like transcription factors in lamprey. AB - The jawless Agnatha (lampreys and hagfishes) represent the phylogenetically oldest order of vertebrates that are believed to lack the adaptive immune system of jawed vertebrates. In order to search for molecular markers specific for cellular components of the adaptive immune system in lampreys, we used the polymerase chain reaction (PCR) to identify genes for transcription factors of the Ikaros family in genomic DNA and cDNA libraries from two species of lampreys, Petromyzon marinus and Lampetra fluviatilis. The mammalian Ikaros-like family of transcription factors consists of five members, Ikaros, Helios, Aiolos, Eos and Pegasus, of which the first three appear to be essential for lymphocyte development. Two different Ikaros-like genes, named IKLF1 and IKLF2, were identified in lamprey. They both have the conserved exon-intron structure of seven exons and show alternative splicing like their counterparts in jawed vertebrates. The genes code for predicted proteins of 589 and 513 amino acid residues, respectively. The proteins contain six highly conserved zinc finger motifs that are 83-91% identical to the mammalian members of the Ikaros-like family. The remaining parts of the sequences are, however, mostly unalignable. Phylogenetic analysis based on the alignable segments of the sequences does not identify the orthologous gene in jawed vertebrates but rather shows equidistance of the lamprey Ikaros-like factors to each other and to Ikaros, Helios, Aiolos and Eos. Expression studies by reverse transcription (RT)-PCR and in situ hybridization (ISH), however, provide evidence for moderate expression in presumed lymphoid tissues like the gut epithelium and for high levels of expression in the gonads, especially in the ovary. PMID- 11896934 TI - Degranulation of primary and secondary granules in adherent human neutrophils. AB - The aim of this study was to investigate whether neutrophil adhesion to extracellular matrix proteins like fibronectin, fibrinogen, and albumin influence the release proteins from primary and secondary granules of neutrophils stimulated by phorbol-myristate-acetate (PMA) and formyl-methionyl-leucyl phenylalanine (f-MLP). Isolated granulocytes plated on wells coated with fibronectin, fibrinogen, and albumin were stimulated with f-MLP (10-7 mol/l), PMA (10-9 mol/l), Mn2+ (5 mmol/l), or combinations of these stimuli, and the degree of adhesion to protein-coated surfaces and the amount of granule proteins released was quantified during 90 min of incubation. PMA, in combination with Mn2+, induced a maximum release of approximately 80% of the intracellular content of lactoferrin and human neutrophil lipocalin (HNL) and 15-20% of the myeloperoxidase (MPO) content regardless of the protein used. PMA or f-MLP alone induced 30-40% release of lactoferrin and HNL depending on the protein that the cells were plated on. Adhesion and release of lactoferrin and HNL were quantitatively related when induced by PMA and PMA plus Mn2+, but not by f-MLP. The mean release of lactoferrin and HNL showed a significant negative relationship to the viability of the cells. In conclusion, adhesion modulates neutrophil degranulation, but it is not always quantitatively related or related in time. PMID- 11896933 TI - IFN-gamma regulates murine interferon-inducible T cell alpha chemokine (I-TAC) expression in dendritic cell lines and during experimental autoimmune encephalomyelitis (EAE). AB - Murine interferon-inducible T cell alpha chemokine (I-TAC) is a potent non-ELR Cys-X-Cys (CXC) chemokine that predominantly attracts activated T lymphocytes and binds to the receptor CXCR3. Using semiquantitative reverse transcription polymerase chain reaction (RT-PCR) we analysed murine I-TAC expression in two different progenitor dendritic cell (DC) lines, MTHC-D2 and JAWS II which were exposed to various cytokines, and Con A-activated splenocytes from a panel of knockout mice. Analysis of the progenitor DC lines and Con A cultures demonstrated that murine I-TAC is primarily regulated by interferon (IFN)-gamma via interferon regulatory factor (IRF)-1. It has been proposed that I-TAC may have a role in autoimmune diseases such as multiple sclerosis (MS). Because I-TAC appears to be secreted from antigen-presenting cells (APCs) and attracts activated T cells, we examined the level of murine I-TAC mRNA in the central nervous system (CNS) of wild-type and IFN-gamma-receptor knockout (IFN-gammaR-/-) mice with myelin oligodendrocyte glycoprotein (MOG)35-55 peptide-induced experimental autoimmune encephalomyelitis (EAE). Peak I-TAC expression was detected in wild-type mice on day 14 when the mice begin to recover, whereas very low levels of I-TAC were detected in the CNS of IFN-gammaR-/- mice which develop severe EAE and die. The expression characteristics of murine I-TAC suggest an important mediator of immune cell communication that could augment vaccines and autoimmune therapies. PMID- 11896935 TI - Contribution of HLA class I allele expression to CD8+ T-cell responses against Epstein-Barr virus. AB - Most immune responses to viral infections involve CD8+ T cells recognizing viral peptides of typically 9-10 amino acids in the groove of major histocompatibility complex (MHC) class I. Importantly, CD8+ T-cell responses appear to focus on few viral epitopes, a phenomenon termed immunodominance. While the understanding of this phenomenon has been based largely on experimental mice models, it is imperative to evaluate its contribution in humans, as the design of peptide-based vaccines may be influenced by immunodomination. Here, we present evidence that immunodominance can be detected among Epstein-Barr virus (EBV) epitopes associated with two of the most frequent class I alleles in Western Europe, human leucocyte antigen (HLA)-A2 and HLA-B7. CD8+ T-cell responses to HLA-A2-associated EBV epitopes were significantly reduced in individuals coexpressing HLA-B7. The impairment of HLA-A2-associated responses correlated with a dominant response to an HLA-B7 epitope. The data demonstrate a hierarchy in the human cellular immune response to immunodominant EBV epitopes presented by separate HLA class I alleles. This may have implications for EBV vaccine development as well as for the interpretation of isolated analysis of immunodominant responses to EBV. PMID- 11896936 TI - Binding of the Galanthus nivalis agglutinin to thymocytes reveals alterations in surface glycosylation during T-cell development. AB - Surface binding of the Galanthus nivalis agglutinin (GNA) to thymocyte subsets has been studied in pigs and rodents by multicolour flow cytometry. In all the species examined, analogous staining profiles have been recorded. Counter staining with anti-CD3epsilon, anti-CD4 and anti-CD8 monoclonal antibodies (MoAb) revealed that a significant increase of the GNA targets on the cell surface occurred during early thymocyte differentiation and reached its maximum at the level of the CD3loCD4+CD8+ small cortical thymocyte. This was followed by a decrease in the GNA binding capacity upon terminal maturation to the single positive thymocytes. PAGE analysis has revealed a dominant GNA-binding glycoprotein (molar mass approx. 90 kDa) present on thymocyte plasma membranes and absent on the surface of splenic lymphocytes, although both the whole cell lysates from both organs contained GNA ligands of the same size. Our findings are in agreement with previous data showing that immature thymocytes differ from their mature counterparts and peripheral T lymphocytes in the surface glycosylation pattern, and support the hypothesis that lectin-glycoprotein interaction plays a significant role in the cell-to-cell crosstalk in the thymic cortex. PMID- 11896937 TI - Specific proliferative and antibody responses of premature infants to intestinal colonization with nonpathogenic probiotic E. coli strain Nissle 1917. AB - The aim of this study was to analyze the influence of oral administration of E. coli Nissle 1917 on the systemic humoral and cellular immunity in premature infants. Thirty-four premature infants were colonized with E. coli Nissle 1917 in a randomized, placebo-controlled blinded clinical trial. Stool samples of infants were analyzed repeatedly for the presence of the administered strain. The proliferative response to bacterial antigens of E. coli origin was measured in whole blood of 34 colonized infants and 27 noncolonized controls. E. coli colonization induced a significant increase in the proliferation of blood cells cultivated with bacterial components of E. coli Nissle 1917 and another E. coli strain in colonized infants as compared with noncolonized controls. Significantly higher amounts of specific anti-E. coli Nissle 1917 antibodies (Ab) of immunoglobulin (Ig)A isotype and nonspecific polyclonal IgM were found in the blood of colonized infants compared to noncolonized placebo controls. We concluded that the oral application of E. coli Nissle 1917 after birth significantly stimulates specific humoral and cellular responses and simultaneously induces nonspecific natural immunity. PMID- 11896939 TI - Ultrasound and left-handedness: a sinister association? PMID- 11896938 TI - The effect of LPS on neutrophils from patients with high risk of type 1 diabetes mellitus in relation to IL-8, IL-10 and IL-12 production and apoptosis in vitro. AB - Innate immunity includes neutrophil inflammatory function, tissue destruction and regulatory cytokine production. Programmed cell death (apoptosis) is postulated to be a key mechanism for neutrophil elimination during inflammation. The aim of the present study was to evaluate the neutrophil apoptosis in relation to IL-8, IL-10 and IL-12 production in vitro by neutrophils of patients suffering from diabetes mellitus (DM)1 and the first-degree relatives of patients with DM1. The early stage of neutrophils apoptosis was assessed morphologically, and the later stage by DNA-binding dye propidium iodide, both after treatment with lipopolysaccharide (LPS), insulin or anti-CD95 antibody (Ab) as stimulators. CD16 (FcgammaRIII) receptor expression was also evaluated. Production of IL-8, IL-10, and IL-12 cytokine was evaluated in supernatant after neutrophil incubation for 21 h in culture medium alone, in medium in the presence of LPS, insulin or anti CD95 antibody (Ab). Cytokine concentrations were measured by enzyme-linked immunosorbent assay (ELISA) method using commercially available kits. Our study demonstrates that LPS inhibits the early stage of apoptosis (as evaluated morphologically) of healthy donors' neutrophils. The LPS-dependent early apoptosis inhibition of neutrophil of patients with DM1 or in prediabetics was decreased in comparison with control. The later stage of apoptosis of neutrophils treated in vitro with anti-CD95 Ab of patients suffering from DM1 was decreased in comparison with prediabetics and healthy donors (propidium iodide (PI) staining). LPS-induced production of anti-apoptotic cytokines IL-8, IL-10 by neutrophils of prediabetic and patients with DM1 was increased. The formyl methionyl-leucyl-phenylalanine (fMLP)-induced proapoptotic reactive oxygen intermediates (ROI) production was significantly higher in DM1 patients. We have concluded that neutrophils from prediabetic and diabetic patients demonstrated the misbalance in anti-apoptotic IL-8 and IL-10 cytokine and proapoptotic ROI production. LPS-dependent IL-12 overproduction by neutrophils is responsible for the switch in T helper Th1/Th2 balance to Th1 and in this way may participate in inflammation and autoimmune DM1 progression. PMID- 11896940 TI - The role of ultrasound scanning on the labor ward. PMID- 11896941 TI - Uterine and fetal cerebral Doppler predict the outcome of third-trimester small for-gestational age fetuses with normal umbilical artery Doppler. AB - OBJECTIVE: To assess the value of different admission tests in predicting the outcome of small-for-gestational age (SGA) fetuses with normal Doppler waveforms in the umbilical artery. METHODS: Criteria for admission into this retrospective study included: singleton pregnancy with a birth weight < 10th centile; absence of severe maternal complications; no evidence of fetal anomalies on the sonogram; normal umbilical artery Doppler; and availability of complete follow-up. At the first antenatal sonogram classifying the fetus as SGA, Doppler analysis of the uterine and middle cerebral arteries was performed and amniotic fluid volume was assessed. Outcome variables included adverse perinatal outcome (perinatal death, severe morbidity) and emergency Cesarean section for fetal distress. RESULTS: Two hundred and thirty-one pregnancies were included in the study. The mean +/- standard deviation birth weight and gestational age at delivery were 2222 +/- 502 g and 37.3 +/- 2.9 weeks, respectively. In 37 cases (16%), an emergency Cesarean section was performed. There was one intrauterine death and three fetuses delivered by emergency Cesarean section developed severe morbidity. Logistic regression demonstrated that abnormal velocimetry of the uterine arteries and fetal middle cerebral artery were independently correlated with the occurrence of Cesarean section. CONCLUSIONS: SGA fetuses with normal umbilical artery Doppler waveforms and abnormal uterine arteries and fetal middle cerebral artery waveforms have an increased risk of developing distress and being delivered by emergency Cesarean section. Particularly when both uterine and fetal cerebral waveforms are altered at the same time, the risk is exceedingly high (86%) and delivery as soon as fetal maturity is achieved seems advisable. On the other hand, when both vessels have normal waveforms, the chances of fetal distress are small (4%) and expectant management is the most reasonable choice. PMID- 11896942 TI - First-trimester placental volume as a marker for chromosomal anomalies: preliminary results from an unselected population. AB - OBJECTIVE: To compare first-trimester placental volume in chromosomally abnormal and normal pregnancies. METHODS: Placental volumes were routinely recorded at the time of nuchal translucency thickness measurement at 10-13 weeks of gestation. This was done using customized three-dimensional ultrasound equipment and measurements were then converted to the placental quotient (placental volume/fetal crown-rump length). The possible difference in placental quotient between chromosomally normal and abnormal pregnancies was examined. RESULTS: A total of 2863 pregnancies was evaluated, including 17 with major chromosomal defects (nine cases of trisomy 21, four of trisomy 18, two of trisomy 13, and one each of Turner syndrome and 48,XXY + 21). The median placental quotient in the chromosomally abnormal group (0.67) was significantly lower than that in the normal fetuses (0.98). In nine of the 17 affected pregnancies the quotient was below the 10th centile of the normal range. CONCLUSIONS: Assessment of placental volume may prove to be useful in first-trimester risk assessment for chromosomal anomalies. PMID- 11896943 TI - Short-term safety of celocentesis for the mother and the fetus. AB - OBJECTIVE: Celocentesis offers the potential for prenatal diagnosis from as early as 6 weeks of gestation. The aim of this study was to examine the short-term safety of celocentesis. METHODS: Eligible for the study were pregnant women with single live fetuses at 6-10 weeks of gestation, requesting pregnancy termination for social indications. At presentation, the patients were asked if they were willing to undergo celocentesis and in those women who agreed the procedure was performed at the time of the initial scan. A second scan was carried out just before termination to measure fetal crown-rump length and heart rate. RESULTS: Four hundred and forty-seven women requested termination of pregnancy and 108 of these agreed to have celocentesis. There were no significant differences between the groups in maternal age, prevalence of primigravidas, cigarette smokers, existence of uterine fibroids, the median fetal crown-rump length or the interval between the initial scan (or celocentesis) at presentation and the pregnancy termination. At the time of termination, ultrasound examination demonstrated fetal death in five (4.7%) of the celocentesis group and nine (2.7%) of the controls (odds ratio, 1.804; 95% confidence interval, 0.5912-5.504). In all other cases, there was normal fetal growth and there were no significant differences between the groups in fetal crown-rump length. CONCLUSIONS: The procedure related fetal loss associated with celocentesis may be approximately 2%. PMID- 11896944 TI - Prenatal detection of ductal-dependent congenital heart disease: how can things be made easier? AB - OBJECTIVE: To improve the detection of ductal dependence in fetuses with severe anomalies of the outflow tracts by observing, with directional power Doppler, reverse flow through the aortic arch or ductus arteriosus in a transverse view of the upper mediastinum. METHODS: A slight cranial move of the ultrasound beam from the three-vessel view allows the transverse view of the aortic arch and ductus arteriosus to be visualized simultaneously. This view is orthogonal to the fetal body axis and parallel to the plane of the four-chamber view. In normal fetuses, directional power Doppler interrogation at this level identifies forward flow in both oblique vessels. RESULTS: We examined 43 fetuses with cardiac defects. In five of the cases, there was reversed flow in the aortic arch or ductus arteriosus in addition to severe anomalies of the outflow tracts, including four with hypoplastic left ventricle and one with pulmonary atresia. CONCLUSIONS: Prenatal detection of reversed flow in the aortic arch or ductus arteriosus is associated with complex congenital heart disease with major diminution of forward flow to the corresponding great vessels. PMID- 11896945 TI - The 'tulip sign': a sonographic clue for in-utero diagnosis of severe hypospadias. AB - OBJECTIVE: To describe a unique sonographic sign for prenatal detection of severe penoscrotal hypospadias. METHODS: Ultrasound findings of hypospadias diagnosed in the second trimester of pregnancy in seven patients were compared with postnatal clinical features. All patients were recruited from routine sonographic examinations performed for various obstetric indications. RESULTS: In six of the seven cases with hypospadias, a severe form of peno-scrotal hypospadias was found. In all six cases, a unique ultrasound feature was observed represented by extreme ventral angulation of the penis, with or without chordee, in a form resembling a tulip flower. This 'tulip' is formed by the ventrally bent penis located between the two scrotal folds. In all six patients, the postnatal pictures of the newborns' genitalia corresponded perfectly to the prenatal sonograms. Associated anomalies occurred in two cases, with Nager syndrome in one case and mild renal hydronephrosis in the other case. CONCLUSIONS: The 'tulip sign' is a specific ultrasonic finding of severe hypospadias. The recognition of this may help to distinguish between severe hypospadias and other genital abnormalities (e.g. ambiguous genitalia). PMID- 11896946 TI - Transvaginal sonography of the uterine cervix prior to labor induction. AB - OBJECTIVES: To compare the Bishop score and transvaginal sonographic measurement of cervical length for predicting the mode of delivery following medically indicated induction of labor in term patients. METHODS: The study was conducted prospectively in 179 women who required medically indicated induction of labor. Inclusion criteria were singleton pregnancy, gestational age > 37 weeks of amenorrhea, cephalic presentation and intact fetal membranes. Cervical length was measured upon arrival in the labor room but was not considered when choosing the induction procedure. Two receiver-operating characteristic curves were plotted to calculate the best threshold value for the Bishop score and for cervical length for predicting the risk of Cesarean section. RESULTS: Fifty-three women (29.6%) had a Cesarean section. The Bishop score was not predictive of the delivery mode, although Cesarean section for failure to progress was more frequent when the Bishop score was < or = 5. Among the women with a Bishop score > 5, the cervical length was not predictive of the induction outcome. However, among the women with a Bishop score < or = 5, a cervical length < 26 mm was associated with a lower Cesarean section rate (20.6 vs. 42.9%; P = 0.006). Furthermore, the interval between the beginning of cervical ripening and delivery was shorter in the case of a short cervix (11.01 +/- 6.7 vs. 18.55 +/- 7.07 h; P < 10(-5)). CONCLUSION: The length of the uterine cervix, measured by transvaginal sonography, is a better predictor of the risk of Cesarean section than the Bishop score after induction of labor for medical reasons. In women with an unfavorable Bishop score, a cervical length of < 26 mm is associated with a lower risk of Cesarean section and a shorter duration of labor. PMID- 11896947 TI - Intrapartum fetal head position I: comparison between transvaginal digital examination and transabdominal ultrasound assessment during the active stage of labor. AB - OBJECTIVE: To test the null hypothesis that no correlation exists between transvaginal digital and the gold standard technique of transabdominal suprapubic ultrasound assessments of fetal head position during labor. A secondary objective was to compare the performance of attending physicians vs. senior residents in depicting fetal head position by transvaginal digital examination in comparison with ultrasound, respectively. METHODS: Consecutive patients in active labor at term with normal singleton cephalic-presenting fetuses were included. All participants had ruptured membranes, cervical dilation > or = 4 cm and fetal head at ischial spine station -2 or lower. Transvaginal sterile digital examinations were performed by either senior residents or attending physicians and followed immediately by transverse suprapubic transabdominal ultrasound assessments. Examiners were blinded to each other's findings. Power-analyses dictated number of subjects required. Statistical analyses included Chi-square, Cohen's Kappa test and logistic regression analysis. P < 0.05 was considered statistically significant. RESULTS: One hundred and two patients were studied (n = 102). In only 24% of patients (n = 24), transvaginal digital examinations were consistent with ultrasound assessments (P = 0.002, 95% confidence interval, 16-33). Logistic regression revealed that cervical effacement (P = 0.03) and ischial spine station (P = 0.01) significantly affected the accuracy of transvaginal digital examination. Parity, gestational age, combined spinal epidural anesthesia, cervical dilation, birth weight and examiner experience did not significantly affect accuracy of the examination. The accuracy of the transvaginal digital exams was increased to 47% (n = 48) (95% confidence interval, 37-57) when fetal head position at transvaginal digital examination was recorded as correct if reported within +/- 45 degrees of the ultrasound assessment. The rate of agreement between the two assessment methods for attending physicians vs. residents was 58% vs. 33%, respectively (P = 0.02) with the +/- 45 degrees analysis. CONCLUSIONS: Using ultrasound assessment as the gold standard, our data demonstrate an overall high rate of error (76%) in transvaginal digital determination of fetal head position during active labor, consistent with the null hypothesis. Attending physicians exhibited an almost two-fold higher success rate in depicting correct fetal head position by physical examination vs. residents in the +/- 45 degrees analysis. Intrapartum ultrasound increases the accuracy of fetal head position assessment during active labor and may serve as an educational tool for physicians in training. PMID- 11896948 TI - Intrapartum fetal head position II: comparison between transvaginal digital examination and transabdominal ultrasound assessment during the second stage of labor. AB - OBJECTIVE: To test the null hypothesis that no correlation exists between transvaginal digital examination compared with the gold standard technique of transabdominal suprapubic ultrasound assessment of fetal head position during the second stage of labor. A secondary objective was to compare the performance of attending physicians vs. senior residents in depicting fetal head position by transvaginal digital examination in comparison with ultrasound assessment. METHODS: Consecutive patients in the second-stage of labor at term with normal singleton cephalic-presenting fetuses and ruptured membranes were included. Transvaginal digital examinations were performed by either attending physicians or senior residents and were followed immediately by transverse suprapubic transabdominal sonographic assessments performed by a single sonographer. Examiners were blinded to each other's findings. Power analysis dictated sample size. Exact binomial confidence intervals around observed rates were compared with chi 2 and Cohen's kappa-tests. Logistic regression was applied. P < 0.05 was considered significant throughout. RESULTS: One hundred and twelve patients were studied. The absolute error of transvaginal digital examinations was recorded in 65% of patients (95% confidence interval, 56-74%). Parity, pelvic station, combined spinal epidural anesthesia, length of first or second stages of labor, use of oxytocin augmentation, gestational age, mode of delivery, birth weight, and examiner experience did not significantly affect examination accuracy. Stratification, when the transvaginal digital examination was recorded as correct if occurring within +/- 45 degrees of the ultrasound assessment, reduced the error of the transvaginal digital examinations to 39% (95% confidence interval, 30-49%). Independent variables again did not affect examination accuracy in this assessment modality. Rates of agreement between the two methods for attending physicians compared with residents were not significantly different. The overall degrees of agreement were 40% (95% confidence interval, 26-55%) and 68% (95% confidence interval, 53-80%) (kappa = 0.25 and 0.30) for the absolute agreement and +/- 45 degrees assessment modalities, respectively, for attending physicians, and 31% (95% confidence interval, 20-44%) and 55% (95% confidence interval, 42 68%) (kappa = 0.14 and 0.12) for senior residents. CONCLUSION: Using ultrasound assessment as the gold standard, our data demonstrate a high rate of error (65%) in transvaginal digital determination of fetal head position during the second stage of labor. The performance of senior residents in transvaginal digital examinations did not differ significantly from that of attending physicians. Intrapartum ultrasound increases the accuracy of fetal head position assessment during the second stage of labor. PMID- 11896949 TI - A new guidance system for freehand, obstetric ultrasound-guided procedures. AB - OBJECTIVE: To assess the performance of the UltraGuide 1000 system, and to compare ultrasound-guided freehand mid-trimester amniocentesis with and without the new guidance system. METHODS: One hundred and sixty-nine women referred for mid-trimester genetic amniocentesis were divided into two groups: a control group of 99 women who underwent the procedure by the freehand technique with scored needles and 70 patients who had the procedure carried out with the aid of a guidance system (UltraGuide 1000) with non-scored needles. The procedures were compared for duration, number of punctures and repositionings of the needle, the visibility of the needle during the puncture and the number of bloody taps. RESULTS: The study group had significantly lower rates of reinsertion (none vs. 7.1%), repositioning (7.1% vs. 17.7%), bloody taps (none vs. 6.1%), touching the fetus (5.7% vs. 22.2%) and prolonged duration of the procedure (4.3% vs. 14.4%) compared with the control group. There was one fetal loss in the control group. Non-visibility of the needle before reaching the amniotic sac occurred in 18.6% of cases in the study group and in 38.4% of cases in the control group. CONCLUSIONS: The new guidance system combines the benefits of an attached guide with the flexibility of the 'freehand' technique. Use of the new guidance system for mid-trimester genetic amniocentesis increases needle visibility and lowers the incidence of common complications. PMID- 11896950 TI - Teaching ultrasound-guided invasive procedures in fetal medicine: learning curves with and without an electronic guidance system. AB - OBJECTIVE: To compare the learning curves of inexperienced junior obstetrics/gynecology registrars for ultrasound-guided invasive procedures on a training model, with and without an electronic guidance system. STUDY DESIGN: Four junior registrars performed their first 100 procedures on a training model with a new electronic guidance system, and four other junior registrars performed their first 100 procedures on the same training model without using the guidance system. All procedures were performed using a free-hand technique. We evaluated the quality of the procedure, which we defined as the time spent with the entire needle clearly visualized on the screen over the total duration of the procedure. We constructed learning curves for the eight junior registrars for comparative analysis. RESULTS: Quality of the procedure increased over time for all trainees. The learning curves were significantly steeper for trainees using the electronic guidance system. Trainees using the electronic guidance system performed better in the middle of their learning curve (procedures 25-75). All trainees reached the same level of quality by the end of their 100 procedures. CONCLUSIONS: The automated electronic guidance system helps faster learning but, after 100 procedures on a training model, both groups reached the same level of quality. PMID- 11896951 TI - Down-up sequential separation of the placenta. AB - OBJECTIVE: To characterize the patterns of placental separation during the third stage of labor. METHODS: Continuous real-time ultrasound was performed during the third stage of labor in 101 normal deliveries. The sequence of placental separation was recorded for determining whether the process was multiphasic, the site from which separation commenced and the mode of its progression. RESULTS: Separation in 97 cases was multiphasic. Monophasic separation in which all parts of the placenta appeared to separate simultaneously occurred in two cases only. Pathological prolongation of the third stage precluded determination of separation in two cases. Ninety-two cases had a uterine wall placenta (anterior or posterior); the separation commenced at one pole and progressed sequentially towards the opposite side in 89 of them. The process started at the lower pole (down-up separation) in 83/92 cases (90.2%) and began from the upper pole (up down separation) in only 6/92 cases (6.5%). Nine cases had a fundal placenta; of these the separation was also multiphasic but began sequentially from either the anterior or posterior pole, or simultaneously from both, in 8 (88.9%) cases so that the fundal part was separated last (bipolar separation). CONCLUSIONS: Placental separation is usually an orderly multiphasic phenomenon that begins mostly from the lower pole of the placenta and propagates sequentially upwards. Fundal placentae, however, separate first at their poles with the fundal part being separated last. Recognition of the sequence of events and understanding of the mechanism of placental separation may aid in detecting cases prone to third stage complications and in managing pathological ones. PMID- 11896952 TI - Reproducibility of three-dimensional ultrasound endometrial volume measurements in patients with postmenopausal bleeding. AB - OBJECTIVE: To evaluate the reproducibility of transvaginal three-dimensional (3D) endometrial volume measurement in patients with postmenopausal bleeding and to compare the reproducibility of this technique to that of two-dimensional (2D) endometrial thickness measurement. METHODS: In a prospective, blinded study, transvaginal ultrasound examinations were performed in 51 consecutive patients with postmenopausal bleeding. Three-dimensional volume and 2D thickness measurements were made and intraobserver and interobserver reproducibility of each technique were assessed. RESULTS: The intraobserver correlation of 3D endometrial volume measurement of the first observer was 0.97 and that of the second observer was 0.99. Thus, mean intraobserver correlation was 0.98. The mean interobserver correlation was 0.95 (0.95 vs. 0.96). There was no significant difference in reproducibility at different volume cut-offs. The mean intra- and interobserver correlation of endometrium volume measurements for five patients with endometrial carcinoma did not differ significantly from that for patients without carcinoma (0.98, 0.98 vs. 0.98, 0.95). The intraobserver correlation of 2D endometrial thickness measurements from the first observer was 0.71 and that from second observer 0.87. Thus, mean intraobserver correlation of the endometrial thickness measurements was 0.79. The mean interobserver correlation was 0.76 (0.84 vs. 0.70). CONCLUSIONS: Endometrial volume and thickness measurements by 3D and 2D ultrasound, respectively, show good reproducibility but the reproducibility of 3D ultrasound is better. PMID- 11896953 TI - First-trimester diagnosis of fetal hepatic cyst. AB - Fetal intra-abdominal cysts seen on antenatal sonography pose a diagnostic problem as they may have many etiological origins. We present a case of a hepatic cyst measuring 11 x 7 x 7 mm that was diagnosed at 13 weeks' gestation by transvaginal sonography. The cyst increased in proportion with the growth of the fetus. Ultrasound-guided needle aspiration of the cyst at 22 weeks' gestation helped to clearly identify the formerly displaced gall bladder and demonstrated the intrahepatic location of the cyst. The aspirated fluid was identified as bile. After aspiration the fluid reaccumulated rapidly. Shortly prior to delivery the cyst measured 75 x 44 x 46 mm. At 39 weeks of gestation a female infant was delivered by forceps (3610 g; Apgar 9/10/10 at 1, 5 and 10 min, respectively). Increasing cyst size and concomitant feeding problems prompted surgery on the 14th day postpartum. A large hepatic cyst was partially excised and marsupialized, confirming the prenatal diagnosis. The postoperative course was complicated by cholangitis, septicemia and recurrence of the cyst. Therefore Roux en-Y hepatojejunostomy was performed in the second month of life. The postoperative period was uneventful and the child was doing well at the time of writing. PMID- 11896954 TI - Sonographic diagnosis of spina bifida at 12 weeks: heading towards indirect signs. AB - Retrospective examination of ultrasound images obtained at 12 weeks of gestation in two fetuses with spina bifida demonstrated retraction of the frontal bones, resulting in an acorn-shaped head, the cerebral peduncles appearing parallel to each other. These craniocerebral signs may improve the accuracy of first trimester diagnosis of spina bifida by sonography. PMID- 11896955 TI - Intrauterine death of one twin, with rescue of the other, in twin-twin transfusion syndrome. AB - Single intrauterine death may occur in twin-twin transfusion syndrome. We investigated why the outcome of the surviving twin is fairly good when the donor twin dies first compared with when the recipient twin dies first. A detailed hemodynamic study was performed using Doppler ultrasound in a twin pregnancy affected by twin-twin transfusion syndrome before and after a single intrauterine death that occurred in the donor twin at 26 weeks' gestation. The recipient twin was expected to die due to severe right cardiac failure with functional stenosis of the pulmonary artery 2 days before the cotwin's death. The donor twin's death caused a prompt resolution of cardiac failure and improvement in other indices, including flow velocity waveform patterns of the umbilical vein, the middle cerebral artery and the ductus venosus. A healthy, premature female neonate weighing 1630 g with a hemoglobin concentration of 17.8 g/dL was delivered by Cesarean section following rupture of the fetal membranes 28 days after the episode. Hemorrhaging from the surviving twin to the dead twin that occurred just before or after the cotwin's death may have contributed to the decrease in volume overload in the recipient twin, leading to a prompt amelioration of the critical hemodynamic indices. The early death of the donor twin may thus have played a significant role in improving the status of the recipient twin in this case of twin-twin transfusion syndrome. PMID- 11896956 TI - Cervical ectopic pregnancy: a case report and literature review. AB - We report a case of cervical ectopic pregnancy that was diagnosed using transabdominal ultrasound. Conservative management with methotrexate administration was undertaken and, following a period of heavy bleeding, bilateral uterine artery embolization was performed. Two weeks after presentation, the gestational sac was shown to have reduced in size. We describe the ultrasound findings in this case and discuss those reported in the literature along with the available management options. PMID- 11896957 TI - Transvaginal sonographic examination of the cervix in asymptomatic pregnant women: review of the literature. AB - Different strategies have been developed to refine the prediction of the risk of preterm delivery in asymptomatic patients. Transvaginal sonography has been used for this reason to measure and examine the length and shape of the cervix. In this review, we focus on clinical studies involving transvaginal sonographic assessment of the cervix in asymptomatic women at high risk of preterm delivery and in the general pregnant population. Three ultrasound signs are suggestive of cervical incompetence, namely, dilatation of the internal os, sacculation or prolapse of the membranes into the cervix (with shortening of the functional cervical length) either spontaneously or induced by transfundal pressure, and short cervix in the absence of uterine contractions. Transvaginal sonography has clearly demonstrated that cerclage leads to a measurable increase in cervical length which may contribute to the success of this procedure in reducing the risk of preterm delivery. Several non-randomized interventional studies among patients with cervical incompetence have been published. They have defined a new group of patients requiring cerclage when the women show progressive cervical modifications on transvaginal sonography, while in other studies, cerclage performed on the basis of cervical changes on transvaginal sonography did not prevent premature delivery. One prospective randomized trial in asymptomatic high risk women has shown two benefits of cerclage following indications for transvaginal sonography: (1) it would generate fewer prophylactic cerclages in high-risk women; (2) therapeutic cerclage before 27 weeks may reduce the incidence of premature delivery before 34 weeks. The risk of preterm delivery is inversely correlated with cervical length. Routine transvaginal sonography of the cervix performed between 18 and 22 weeks can help identify patients at risk of preterm delivery. However, given the low prevalence of preterm births, screening would generate either a high false-positive rate or a low sensitivity. One non randomized interventional study among patients with a short cervix on routine ultrasound examination found a lower risk of delivery before 32 weeks in the cerclage group than in the expectant management group. However, to date, there have been no prospective randomized trials in a general population. Although evidence is still lacking, there does appear to be a benefit in performing a cerclage rather than continuing with expectant management in cases with sonographic appearance of cervical incompetence in asymptomatic women at high risk of preterm delivery. Ultrasound can be offered to reduce the indications of cerclage for cases in which the situation is uncertain. Within the general obstetric population, transvaginal sonography might help in the selection of asymptomatic but high-risk women. However, the benefit associated with cerclage for sonographic indication has not been demonstrated. PMID- 11896958 TI - The association of spinal muscular atrophy type II and increased nuchal translucency. PMID- 11896959 TI - Occlusion of umbilical artery using a Guglielmi detachable coil for the treatment of TRAP sequence. PMID- 11896960 TI - Re: Fetal urine production and accuracy when estimating fetal urinary bladder volume. PMID- 11896961 TI - The 'tulip sign': a sonographic clue for in-utero diagnosis of severe hypospadias. PMID- 11896962 TI - Literature review by the ISUOG Bioeffects and Safety Committee. PMID- 11896964 TI - Primary cell culture and morphological characterization of canine dermal papilla cells and dermal fibroblasts. AB - Skin biopsies were taken from female dogs, the primary hair follicles isolated and the dermal papilla dissected. After incubation in supplemented Amniomax complete C100 medium in 24-well culture plates, the dermal papilla cells (DPC) grew to confluence within 3 weeks. Thereafter, they were subcultivated every 7 days. Dermal fibroblast (DFB) cultures were established by explant culture of interfollicular dermis in serum-free medium, where they reached confluence in 10 days. They were subcultivated every 5 days. For immunohistochemistry, cells were grown on cover slips for 24 h, fixed and stained with antibodies against collagen IV and laminin. DPC showed an aggregative growth pattern and formation of pseudopapillae. Intensive staining for collagen IV and laminin could be observed until the sixth passage. DFB grew as branching, parallel lines and showed only weak staining for collagen IV and laminin. PMID- 11896965 TI - Retrospective study: the presence of Malassezia in feline skin biopsies. A clinicopathological study. AB - Malassezia spp. dermatitis, a rare disorder in cats, has previously been associated with immune suppression and internal malignancies. This study evaluates the presence and importance of Malassezia spp. in feline biopsy specimens submitted for histopathological examination. Five hundred and fifty haematoxylin and eosin-stained skin biopsy specimens received for histopathological examination between January 1999 and November 2000 were reviewed. Fifteen (2.7%) submissions contained Malassezia organisms in the stratum corneum of the epidermis or follicular infundibulum. Eleven of 15 cats presented with an acute onset of multifocal to generalized skin lesions. All 11 cats were euthanized or died within 2 months of the onset of clinical signs. Seven cats had dermatopathological changes and clinical signs supportive of paraneoplastic alopecia, and three cats had an interface dermatitis suggestive of erythema multiforme or thymoma-associated dermatosis. Histopathological changes were nonspecific in one cat that was euthanized 2 weeks following onset of severe pruritus and alopecia. In three cats, Malassezia spp. were found in localized sites (two chin, one footpads) and appeared inconsequential to their overall health status. One cat had Malassezia spp. in association with cutaneous demodicosis. These findings suggest that Malassezia yeast in dermatopathological specimens from multifocal or generalized lesions should prompt a thorough clinical work-up for internal neoplasia. PMID- 11896966 TI - Haemangiopericytoma: histological spectrum, immunohistochemical characterization and prognosis. AB - Canine haemangiopericytoma (CHP) is a vascular neoplasm thought to be derived from pericytes. The histological pattern and immunohistochemical profile were studied in 31 CHPs. Twenty-three subjects were followed for 2 years to evaluate the correlation among tumour location, histotype, immunostaining and outcome of the disease. Of the 31 CHPs examined, 20 exhibited a perivascular whorled pattern, 8 were storiform and 3 were epithelioid. All tumours were positive for vimentin and negative for cytokeratin, factor VIII-related antigen, glial fibrillary acidic protein and S-100 protein. Seventeen CHPs were positive for actin and nine co-expressed desmin. Six CHPs were also positive for CD34 antigen. The panel of immunohistochemical markers used confirmed the vascular lineage of CHP and aided in the exclusion of other mesenchymal tumours. Of the 23 dogs submitted to follow-up, 6 had recurrence or metastases of the primary tumour. The epithelioid pattern or a noncutaneous location were associated with a poorer prognosis. PMID- 11896967 TI - Evaluation of topically applied enilconazole for the treatment of dermatophytosis in a Persian cattery. AB - Many Persian catteries have long-standing dermatophyte infections and are particularly difficult to treat. Enilconazole is a topical antifungal agent that has demonstrated good efficacy in recent studies. Twenty-two Persian cats naturally infected with Microsporum canis in a breeding cattery were treated with topical 0.2% enilconazole and monitored for 180 days. The treatments were repeated every 3 days for a total of eight applications. All the cats improved clinically and became culture negative by day 28. By day 180, four cats had developed clinical dermatophytosis and all cats had positive fungal cultures. In this study, topical 0.2% enilconazole was generally well tolerated but may have caused hypersalivation, idiopathic muscle weakness and slightly elevated serum alanine aminotransferase (ALT) concentrations. This study suggests that enilconazole may be used safely with little risk to the young, aged and gravid animals. PMID- 11896968 TI - Use of computerized image analysis to quantify staphylococcal adhesion to canine corneocytes: does breed and body site have any relevance to the pathogenesis of pyoderma? AB - An optimized system of computerized image analysis was used to investigate variations in the adherence of Staphylococcus intermedius to canine corneocytes from four different breed groups and six different anatomical sites. S. intermedius showed significantly greater adherence to the head and neck compared with the dorsum, but adherence to the limb, axilla and groin did not differ from other sites. Furthermore, there was significantly greater adherence of S. intermedius to corneocytes from the dorsum, forelimb, axilla and groin of Boxers and Bull Terriers than Spaniels and Hounds. S. intermedius, and also Pseudomonas aeruginosa, exhibited abundant adherence, which was significantly greater than Staphylococcus aureus, Streptococcus canis, Klebsiella pneumoniae and Escherichia coli. In addition, S. intermedius adherence demonstrated a sigmoid dose-response curve with increasing bacterial concentration. These results suggest that S. intermedius adheres to canine corneocytes by a specific receptor-ligand interaction and adheres to the skin of some breeds more avidly than others. However, variations in adherence between body regions would not account for the predilection sites of canine bacterial pyoderma. PMID- 11896969 TI - Sensitivity patterns to house dust mites and forage mites in atopic dogs: 150 cases. AB - This study investigated intradermal test reactions to extracts of six species of mites in 150 dogs with atopic dermatitis. At least one positive reaction was seen in 120 animals (80%). Dermatophagoides farinae attracted the highest number of positive reactions (108 dogs, 90% of dogs and 72% of atopic dogs showing positive reactions). Positive reactions to other mites were not uncommon, with many dogs testing positive for Dermatophagoides pteronyssinus (32% of dogs tested), Acarus siro (35%), Tyrophagus putrescentiae (30%), Glycyphagus domesticus (27%) and Lepidoglyphus destructor (23%). Sensitivity to D. farinae alone occurred commonly (57% of cases), but multiple sensitivities were seen frequently with the other mites. Cases of sensitivity to only one mite were also seen: D. pteronyssinus (five cases), T. putrescentiae (one case) and G. domesticus (one case). Further studies are needed to appreciate more clearly the precise role played by the different species of mite in canine atopic dermatitis. PMID- 11896970 TI - Intracorneal vacuoles in skin diseases with parakeratotic hyperkeratosis in the dog: a retrospective light-microscopy study of 111 cases (1973-2000). AB - Two recent case reports described a congenital keratinization defect (congenital follicular parakeratosis; CFP) in Rottweiler and Siberian Husky dogs. Skin biopsy specimens revealed marked parakeratosis targeting the hair follicle and numerous intracorneal vacuoles. A retrospective histopathological study was conducted on skin biopsy specimens from 111 dogs with diseases associated with parakeratotic hyperkeratosis to determine whether intracorneal vacuoles were present. Additional criteria evaluated were the size and location of the vacuoles and the degree of parakeratosis. Cases examined included dogs with primary idiopathic seborrhoea, necrolytic migratory erythema (NME), Malassezia dermatitis, zinc responsive dermatosis, hereditary nasal hyperkeratosis of Labrador Retriever dogs, thallotoxicosis and CFP. Thirty-seven cases (37/111, 33%) had intracorneal vacuoles, including nine cases of primary idiopathic seborrhoea (9/29, 31%), 10 cases of NME (10/18, 56%), five cases of Malassezia dermatitis (5/19, 26%), five cases of zinc-responsive dermatosis (5/36, 14%), five cases of hereditary nasal hyperkeratosis (5/5, 100%) and three cases of CFP (3/3, 100%). If present, intracorneal vacuoles were found throughout all layers of the parakeratin. The sizes of intracorneal vacuoles varied among diseases, but large (> 5 microm) vacuoles only were present in CFP. Biopsies with a larger degree of parakeratosis were significantly more likely to have intracorneal vacuoles (P = < 0.001). Based on this study, intracorneal vacuoles are a common finding in many parakeratotic skin diseases of the dog, but large (> 5 microm) vacuoles are found only in CFP. PMID- 11896971 TI - Dermatophyte granulomas caused by Trichophyton mentagrophytes in a dog. AB - Multiple, dermal and subcutaneous nodules developed in a young female Manchester Terrier dog that had a chronic history of superficial dermatophytosis. Skin biopsy specimens of the nodules revealed granulomatous inflammation in the deep dermis and subcutis with branching fungal organisms. Cultures of multiple biopsy specimens from the nodules all yielded Trichophyton mentagrophytes. The lesions in this dog were similar to granulomatous dermatophytosis, a skin disease that has been reported in Persian cats and one Yorkshire Terrier dog. PMID- 11896972 TI - Seasonal atopic dermatitis in dogs sensitive to a major allergen of Japanese cedar (Cryptomeria japonica) pollen. AB - Three dogs were examined because of episodes of recurrent pruritic dermatitis in the spring, the season of Japanese cedar (Cryptomeria japonica, CJ) pollination in Japan. The dogs were shown to be sensitive to CJ pollen allergen using intradermal testing and antigen-specific IgE measurement. Fluorometric enzyme linked immunosorbant assay (ELISA) showed increased concentrations of IgE specific to Cry j 1 and a negative result for Cry j 2 in the three dogs. The concentrations of IgE specific to Cry j 1 during the season of CJ pollination were higher than the concentrations found during the off-season in all the dogs, and the variation in the concentrations correlated with the variation in clinical signs. Peripheral blood mononuclear cells showed apparent proliferative responses to crude CJ pollen antigen and Cry j 1 during CJ pollination season. These findings indicated that Cry j 1 was the major allergen recognized by IgE and lymphocytes and resulted in the development of type I hypersensitivity to CJ pollen allergen in these atopic dogs. PMID- 11896973 TI - Zinc-responsive dermatosis in dogs. PMID- 11896975 TI - Use of molecular techniques to study microbial diversity in the skin: chronic wounds reevaluated. AB - The skin is colonized by an array of microorganisms which form its natural microflora. Disruption to the normal barrier function of the skin (due to trauma or disease) may result in invasion of the dermis by opportunistic bacteria. To date, these organisms, which may contribute to the chronicity of skin wounds, have been analyzed solely by culture methods. It is increasingly realized that standard culture methods of analysis do not accurately reflect the full diversity of complex microflora. This review discusses the limitations of traditional culture approaches and reviews recent advances in molecular microbiological techniques which facilitate a more comprehensive characterization of the microflora within clinical samples. The currently available technologies and techniques are described, as is their use in clinical practice and their potential for diagnostic screening. Chronic venous ulceration of the lower limbs is an important skin disorder in which the microflora invading the dermal tissues contribute to the observed delayed healing. Using chronic leg ulcers as a working example, we show how strict culture and molecular microbiological techniques may be employed, for the first time in combination, to definitively characterize the invading microbial community of the dermis. PMID- 11896976 TI - Success and limitations of a naked plasmid transfection protocol for keratinocyte growth factor-1 to enhance cutaneous wound healing. AB - Our group and others have previously reported enhancement of cutaneous wound healing following the transfection of tissue with plasmid vectors expressing the DNA for growth factors. In these experiments, growth factor treated animals were usually compared to animals treated with control plasmid vector. To achieve consistent transfection, high DNA plasmid load and repeated penetrations of the wound by needle or gene gun were required. In the current experiments, we assessed the effect of the plasmid load and repeated tissue penetrations on wound healing of excisional wounds in diabetic C57 mice. Animals received 5 mm excisional wounds, and were assigned to the following groups, no treatment, phosphate buffered saline solution injections, and plasmid vector injection with and without the keratinocyte growth factor-1 gene. Intradermal injections of 100 microg plasmid were given adjacent to the wounds at days 1-5, 7 and 11. At day 9, wound closure was more advanced in keratinocyte growth factor-1 treated animals compared to those treated with control plasmid. But a detrimental effect of the DNA plasmid injection was evident from a comparison of the DNA control group versus the non-injected group. Therefore, the challenge for developing an effective system for the enhancement of wound healing lies in improving transfection efficiency. PMID- 11896977 TI - Randomized trial of topically applied repifermin (recombinant human keratinocyte growth factor-2) to accelerate wound healing in venous ulcers. AB - About 600,000 people in the United States are estimated to be affected by venous ulcers. The cornerstone of care of chronic venous ulcers involves the application of compression bandages. Other therapies include treatment of associated infection, treatment for edema and inflammation, and debridement when necessary. Repifermin, a recombinant human KGF-2 (fibroblast growth factor-10), exerts a proliferative effect on epithelial cells, in vitro and in vivo, and has been shown to accelerate wound healing in several experimental animal models. A randomized, double-blind, parallel-group, placebo-controlled, multicenter study was conducted to evaluate the safety and efficacy of topical repifermin treatment, for 12 weeks, in the healing of chronic venous ulcers in 94 patients. Repifermin was shown to accelerate wound healing, with significantly more patients achieving 75% wound closure with repifermin than with placebo. The treatment effect appeared more marked for a subgroup of patients with initial wound areas < or = 15 cm2 and wound ages of < or = 18 months. A longer duration of treatment (e.g., 26 weeks) may allow better differentiation of the benefit of repifermin compared with placebo, particularly with respect to complete wound closure. The safety assessment showed that repifermin was well tolerated. PMID- 11896978 TI - Role of neuropathy and plasma nitric oxide in recurrent neuropathic and neuroischemic diabetic foot ulcers. AB - Various factors are associated with foot ulceration, including delayed reporting of ulcers, poor glycemic control, and severity of neuropathy. Several studies have looked at the role of nitric oxide in wound healing. However, no studies have examined its role in the occurrence and recurrence of diabetic foot ulceration. In a cross-sectional study we examined the role of neuropathy, retinopathy, nephropathy, and plasma nitric oxide (estimated from plasma nitrite and nitrate) levels in diabetic patients with recurrent and non-recurrent neuropathic and neuroischemic foot ulcers. Patients with recurrent foot ulcers had higher vibration perception threshold values compared to patients with non recurrent foot ulcers (47.4 +/- 5.7 volts versus 39.5 +/- 10.3 volts respectively, P < 0.05). In addition, subjects with recurrent foot ulcers had significantly higher plasma nitric oxide compared to subjects with non-recurrent foot ulcers (46.9 +/- 6.3 microm/L versus 30.2 +/- 2.4 microm/L respectively, P < 0.01). Multivariate logistic regression analysis adjusted for age, sex, hemoglobin A1c, presence of retinopathy and nephropathy, vibration perception threshold, plasma creatinine, and total nitric oxide, indicated that only vibration perception threshold was independently associated with the presence of an ulcer [odds ratio: 1.26 (1.10-1.46); P <0.001)] and the recurrence of foot ulcers [odds ratio: 1.13 (1.01-1.27); P =0.04)]. This study has shown that although plasma nitric oxide is higher in patients with recurrent neuropathic and neuroischemic foot ulcers, severity of neuropathy was the most important factor associated with the development and recurrence of foot ulcers in diabetic patients. PMID- 11896979 TI - Early gene expression profile of human skin to injury using high-density cDNA microarrays. AB - Disturbances in normal wound healing may be traced to perturbations in gene expression following injury. To decipher normal and abnormal genetic responses to cutaneous injury, baseline gene expression of uninjured skin and injured skin must be better defined. Our aim for this study was to determine the gene expression profile of human skin immediately following injury using cDNA microarrays. Samples of normal and injured skin were obtained from 5 healthy females undergoing breast reduction surgery. Specimens of the epidermis and dermis were obtained at 30 minutes and 1 hour after the initial injury. RNA was extracted, reverse transcribed into cDNA and hybridized onto high-density cDNA microarray membranes of 4,000 genes. At 30 minutes, injury resulted in a consistent increase (> 2x) in gene expression of 124 out of 4,000 genes (3%). These genes were primarily involved in transcription and signaling. None of the 4,000 genes were decreased (< 2x) at 30 minutes. At 1 hour only 46 out of the 4,000 genes were increased in expression (1.15%) but 264 out of 4,000 (6.6%) genes were decreased greater than 2 fold, indicating a silencing of many structural genes. We have identified several genes, namely, suppressor of cytokine signaling-1, rho HP1, and BB1, that are highly expressed after injury and may have an unappreciated role in regulating the initial inflammatory response. These data provide an initial high-throughput analysis of gene expression immediately following human skin injury and show the utility and future importance of high-throughput analysis in skin biology and wound repair. PMID- 11896980 TI - Age and growth factors in porcine full-thickness wound healing. AB - It has been recognized that the rate of cutaneous wound healing declines with age, yet the molecular processes that affect this decline remain poorly understood. The purpose of this study was to compare reepithelialization and contraction rates, and growth factor profiles in full-thickness wounds in swine of various ages. Multiple full-thickness excisional wounds were created on the dorsum of 24-month-old (n=2), 4-month-old (n=2), and 2-month-old (n=2) Yucatan Minipigs. The extent of reepithelialization was shown to decrease with increasing age in a manner that was statistically significant among the 2-month-old (79%), 4 month-old (48%), and 24-month-old pigs (22%). Enzyme-linked immunosorbent assay results showed that endogenous vascular endothelial growth factor concentrations in the 2- and 4-month-old animals peaked on day 4, reaching levels of 482 pg/ml and 420 pg/ml, respectively. In the 24-month-old pigs the vascular endothelial growth factor concentration peaked later (day 6), and was present at a lower level (229 pg/ml). On day 4 the vascular endothelial growth factor levels in the older pigs reached only 120 pg/ml, representing a four-fold decrease in concentration compared to the younger pigs. A comparison of platelet-derived growth factor-BB concentrations across the age groups showed similar patterns in the 2- and 4-month-old pigs (peaks of 77 and 91 pg/ml on days 2 and 3, respectively), and levels in the 24-month-old were below the sensitivity level (31.5 pg/ml) of the assay. Transforming growth factor-beta1 levels across the age groups did not differ in a manner that was statistically significant, and all age groups peaked on day 9. Wound contraction showed no statistical differences among the age groups from days 3 to 9. On day 11, however, wound contraction in 2-month old pigs was about 10% faster than in 24-month-old pigs (p < 0.05). These data suggest a possible new algorithm for treating wounds in aged skin, by which exogenous growth factors can be added to the wound microenvironment in doses and at times that match the growth factor profiles observed in wounds made in younger skin. PMID- 11896981 TI - Expression of heat shock proteins in a linear rodent wound. AB - Heat shock proteins (hsps) are ubiquitous and known to be expressed in all organisms. These stress proteins are likely to be induced in the wound environment and may play a critical role in the overall process of wound repair. Linear incisions were made in Sprague-Dawley rats. Serial skin biopsies were taken, the dermis and epidermis were separated and a protein lysate made. The expression of hsp 72, 47, and 32 were analyzed by Western blotting and immunohistochemistry. There were distinct patterns of expression of hsp 72, 47, and 32 in the wound. In unwounded dermis, there was no constitutive expression of any of the heat shock proteins studied. In the epidermis, there was constitutive expression of hsp 32 and 72, but not hsp 47. With wounding, all hsps exhibited increased expression both in the dermis and epidermis. These patterns of protein expression are suggestive of the individual heat shock proteins' molecular function, such as hsp 72's role as an indicator of cellular stress and injury, hsp 47's role in collagen synthesis, and hsp 32's role as an antioxidant. PMID- 11896982 TI - Wound epithelialization deficits in the transforming growth factor-alpha knockout mouse. AB - In vitro, transforming growth factor-alpha is an important factor controlling epithelial cell proliferation and migration. However, the transforming growth factor-alpha knockout mouse has shown no wound epithelialization defect in tail amputation and full-thickness back wounds. To resolve this disparity, we combined a full-thickness head wound and a partial-thickness ear wound on the transforming growth factor-alpha knockout mouse for analysis of wound epithelialization with or without granulation tissue formation. Three-millimeter ear wounds were made on the transforming growth factor-alpha knockout and heterozygous control mice. Full thickness head wounds were made using a 6-mm trephine on the crown of the skull. In the ear model, transforming growth factor-alpha knockout mice had significantly larger epithelial gaps versus control at post-operative day 3 and 5. Epithelial thickness at the wound edge of transforming growth factor-alpha deficient mice was also depressed at post-operative day 3 and post-operative day 5 compared to control mice. On post-operative day 8, most wounds of both groups were epithelialized. In contrast, no difference in epithelial gap or new granulation tissue was found in the head model. The data support the concept that transforming growth factor-alpha plays a significant early role in wound epithelialization in vivo but its deficit is compensated if accompanied by granulation tissue formation. The data further show the importance of appropriate wound models to address the role of vulnerary factors. PMID- 11896984 TI - Stem cells in mammalian repair and regeneration. PMID- 11896985 TI - Four moral questions for human embryonic stem cell research. AB - Human embryonic stem cell research offers great promise for the treatment of many serious disease conditions in a variety of medical areas, including wound healing. Before this promise can be realized, society and individual researchers, clinicians, and patients will have to answer four ethical questions: 1) Can we ever intentionally destroy a human embryo? 2) Can we benefit from others' destruction of embryos? 3) Can we create an embryo to destroy it? 4) Can we clone human embryos? After outlining the issues raised by each question, the author concludes by indicating his own affirmative answer to each of these questions. PMID- 11896986 TI - Stem cells in regenerative biology and medicine. AB - Embryonic stem cells of the mammalian blastocyst give rise to all the tissue lineages that begin to emerge at gastrulation. They are pluripotent cells and can be propagated in vitro without loss of pluripotency. Many adult tissues harbor cells that do not complete their differentiation program. These cells serve as self-renewing stem cells whose normal fate is to regenerate site-specific tissue, in response to either physiological cell turnover or damage inflicted by injury or disease. Neural, muscle, and bone marrow stem cells possess developmental potency far greater than their normal lineage-restricted fate. The understanding of the biology of stem cells is leading us into an era of regenerative medicine. The growth potential and pluripotency of embryonic stem cells and the developmental plasticity of adult stem cells, particularly those of bone marrow, make them potentially useful for replacing tissues, via transplantation or construction of bioartificial tissues, that either do not regenerate naturally or are damaged beyond their natural capability for regeneration. In addition to these two ways of replacing tissue, a third strategy of regenerative medicine is to stimulate regeneration in vivo from resident stem cells. Before these approaches become clinical reality, however, a number of basic research issues must be resolved, including the revision of our concept of a regeneration competent cell. PMID- 11896987 TI - Directed differentiation of embryonic stem cells: genetic and epigenetic methods. AB - Embryonic stem cells are derived from the inner cell mass of the pre-implantation blastocyst, and can both self-renew and differentiate into all the cells and tissues of the body. The embryonic stem cell is an unsurpassed starting material to begin to understand a critical, largely inaccessible, period of development, as well as an important source of cells for transplantation and gene therapy. Despite their potential, attempts to obtain specific cell types from embryonic stem cells have been only partially successful because many of the growth factor combinations and developmental control genes involved in cell type restricted differentiation are unknown. This article summarizes some of the recent advances in promoting lineage restricted differentiation of embryonic stem cells, focusing on growth factor manipulation, or genetically altering embryonic stem cells to produce a desired phenotype. The two approaches epitomize current scientific concerns regarding the therapeutic use of these cells; genetic alterations will produce more pure cells with the risk of increasing the likelihood of malignant transformation; epigenetic methods for the manipulation of stem cell phenotype are often incomplete and remaining pluripotent cells are likely to form teratomas. As more is known about lineage specification during development, it will be possible to more precisely control cell type specification. PMID- 11896988 TI - Microenvironment and stem properties of bone marrow-derived mesenchymal cells. AB - Adult stem cells are self-renewing, pluripotent, and able to repopulate the tissue in which they reside. Cells endowed with these properties have been isolated from several tissues and an increasing number of reports provide evidence of their ability, following transplantation, to engraft host tissues other than those of their origin. In this setting, interest in the well documented capacity of bone marrow stromal cells to undergo multilineage differentiation is growing. Neural and cardiomyogenic lineages have recently been proposed as additional differentiative pathways of these cells. However, culture conditions and inductive molecules can alter the behavior of bone marrow stromal cells and the microenvironment is critical for proper in vivo delivery. The maintenance of their stem properties and the possibility of reprogramming their commitment is a field of primary interest given the potential use of these cells in regenerative medicine. We discuss here how the microenvironmental cues, and the growth factors that physiologically govern commitment and subsequent differentiation, influence the properties of bone marrow stromal cells and modulate their engraftment into host tissues. PMID- 11896989 TI - The role of progenitor cells in repair of liver injury and in liver transplantation. AB - There are three levels of cells in the hepatic lineage that respond to injury or carcinogenesis: the mature hepatocyte, the ductular "bipolar" progenitor cell, and a putative periductular stem cell. Hepatocytes are numerous, and respond rapidly to liver cell loss by one or two cell cycles but can only produce other hepatocytes. The ductular progenitor cells are less numerous, may proliferate for more cycles than hepatocytes, and are generally considered "bipolar," i.e., they can give rise to biliary cells or hepatocytes. Periductular stem cells are rare in the liver, have a very long proliferation potential, and may be multipotent. Extrahepatic (bone marrow) origin of the periductular stem cells is supported by recent data showing that hepatocytes may express genetic markers of donor hematopoietic cells after bone marrow transplantation. These different regenerative cells with variations in potential for proliferation and differentiation may provide different sources of cells for liver transplantation: hepatocytes for treatment of acute liver damage, liver progenitor cell lines for liver-directed gene therapy, and bone marrow-derived cells for chronic long-term liver replacement. A limiting factor in the success of liver cell transplantation is the condition of the hepatic microenvironment in which the cells must proliferate and set up housekeeping. PMID- 11896991 TI - Bone marrow stem cell and progenitor response to injury. AB - Hematopoietic stem cells represent a long term reservoir of cells to populate blood with multiple formed cells. These hematopoietic stem cells proliferate and mature into lymphoid, erythroid, and myeloid precursor cells, with the balance of these cell populations modulated by major thermal injury, with or without sepsis. Recent studies indicate that thermal injury shifts this balance to favor the monocyte/macrophage lineage at the expense of neutrophil production. The mechanisms for these changes are now being elucidated with the results of clinical importance, because understanding the dynamics of the different precursor pools could be used to identify patients at greater risk for systemic inflammatory sequelae following major thermal injury. PMID- 11896990 TI - Corneal stem cells in review. AB - The cornea provides the eye with protection and the refractive properties essential for visual acuity. The transparent epithelium is highly specialized with basal and stratified squamous cells that are renewed throughout life from a stem cell population. The stem cells are thought to reside at the corneal limbus and may be maintained by a variety of intrinsic and extrinsic factors such as the local environment, survival factors, and cytokines. A number of markers have been localized to the limbus in an attempt to identify stem cells; however, definite stem cell identification remains elusive. During homeostasis and following injury to the corneal epithelium, the limbal stem cells divide to produce daughter transient amplifying cells that proliferate, migrate, and differentiate to replace lost cells. However, this cannot occur if the stem cell population is depleted. Limbal stem cell deficiency then results in corneal re epithelialization by the neighboring conjunctiva, causing pain, poor vision, and even blindness. This review will focus on corneal epithelial stem cells in ocular surface repair and regeneration. The current knowledge of stem cell biology in the corneal epithelium, clinical consequences of stem cell deficiency, and therapeutic strategies aimed at reversing stem cell deficiency will be discussed. PMID- 11896992 TI - Detection of apoptosis in keloids and a comparative study on apoptosis between keloids, hypertrophic scars, normal healed flat scars, and dermatofibroma. AB - Recent studies have suggested that the regulation of apoptosis during wound healing is important in scar establishment and the development of pathological scarring. In this study, we demonstrate that keloid fibroblasts can be identified as apoptotic cells because of their highly condensed chromatin and discrete nuclear fragments. To further reveal the phenomenon of apoptosis, we quantified the number of terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells in surgically resected tissues of keloids (N = 10), hypertrophic scars (N = 10), normal healed flat scars (N = 10), and dermatofibroma (N = 10). The number of TUNEL-positive cells was relatively low, but was significantly higher for the keloid group compared with the normally healed flat scar group (p = 0.004), suggesting reduced cell survival and increased apoptotic cell death in a subpopulation of keloid fibroblasts. Furthermore, the number of TUNEL-positive cells was significantly higher for the keloid group compared with the dermatofibroma group (p = 0.044), suggesting that a subpopulation of keloid fibroblasts may suppress tumorgenicity at a greater rate than dermatofibroma by undergoing cell death. Hypertrophic scars had significantly higher levels of apoptosis than normally healed flat scars (p = 0.033). Therefore, these results suggest that selected fibroblasts in keloids and hypertrophic scars undergo apoptosis, which may play a role in the process of pathological scarring. PMID- 11896993 TI - Perfusion of medium improves growth of human oral neomucosal tissue constructs. AB - Tissue engineering of the oral mucosa may be useful in congenital cleft palate repairs, defects following extirpative oncologic surgery, and periodontal disease. One of the limitations of in vitro growth of oral mucosal constructs is central necrosis of 3-dimensional tissues. We tested the hypothesis that medium perfusion would enhance oral mucosal histogenesis in vitro. Normal human oral keratinocytes were obtained from young to middle-aged adults. Porous 3 dimensional matrices were prepared from collagen and chondroitin sulfate with some crosslinked with glutaraldehyde. Each device was seeded with 5.0 x 10(5) human oral keratinocytes. The seeded matrices were cultured with or without perfusion of medium at 1.3 ml/min. Histologic analysis of samples cultured for 3, 7, or 14 days showed superior viability and proliferation when perfused. At day 7, the average number of cell layers of the neoepithelium of sponges in the perfused culture system (9.4 +/- 1.0) was 88% greater than for the nonperfused culture system (5.0 +/- 0.9, p<0.005). Glutaraldehyde crosslinking did not influence cellular proliferation or the extent of matrix's shrinkage in either culture system. This study shows that medium perfusion enhanced cell viability and proliferation of human oral keratinocytes cultured in porous 3-dimensional matrices. PMID- 11896994 TI - Acceleration of wound contraction and healing with a photocrosslinkable chitosan hydrogel. AB - Application of ultraviolet light irradiation to a photocrosslinkable chitosan aqueous solution resulted in an insoluble, flexible hydrogel like soft rubber within 60 seconds. In order to evaluate its accelerating effect on wound healing, full-thickness skin incisions were made on the backs of mice and subsequently a photocrosslinkable chitosan aqueous solution was added into the wound and irradiated with UV light for 90 seconds. Application of the chitosan hydrogel significantly induced wound contraction and accelerated wound closure and healing compared with the untreated controls. Histological examination also showed an advanced contraction rate on the first 2 days and tissue fill rate on days 2 to 4 in the chitosan hydrogel-treated wounds. Furthermore, in cell culture studies, chitosan hydrogel culture medium supplemented with 5% fetal-bovine serum was found to be chemoattractant for human dermal fibroblasts in an invasion chamber assay using filters coated with Matrigel and in a cell migration assay. Due to its ability to accelerate wound contraction and healing, chitosan hydrogel may become accepted as an occlusive dressing for wound management. PMID- 11896995 TI - Increased fibrovascular invasion of subcutaneous polyvinyl alcohol sponges in SPARC-null mice. AB - The expression of SPARC (secreted protein acidic and rich in cysteine/osteonectin/BM-40) is elevated in endothelial cells participating in angiogenesis in vitro and in vivo. SPARC acts on endothelial cells to elicit changes in cell shape and to inhibit cell cycle progression. In addition, SPARC binds to and diminishes the mitotic activity of vascular endothelial growth factor. To determine the effect(s) of SPARC on angiogenic responses in vivo, we implanted polyvinyl alcohol sponges subcutaneously into wild-type and SPARC-null mice. On days 12 and 20 following implantation, SPARC-null mice showed increased cellular invasion of the sponges in comparison to wild-type mice. Areas of the sponge with the highest cell density exhibited the highest numbers of vascular profiles in both wild-type and SPARC-null animals. The endothelial component of the vessels was substantiated by immunoreactivity with three different markers specific for endothelial cells. Although sponges from SPARC-null relative to wild type mice were populated by significantly more cells and blood vessels, an increase in the ratio of vascular to nonvascular cells was not apparent. No differences in the percentage of proliferating cells within the sponge were detected between wild-type and SPARC-null sections. However, elevated levels of vascular endothelial growth factor were associated with sponges from SPARC-null versus wild-type mice. An increase in vascular endothelial growth factor production was also observed in SPARC-null primary dermal fibroblasts relative to those of wild-type cells. In conclusion, we have shown that the fibrovascular invasion of polyvinyl alcohol sponges is enhanced in mice lacking SPARC, and we propose that increased levels of vascular endothelial growth factor account, at least in part, for this response. PMID- 11896996 TI - Neonatal rat cartilage has the capacity for tissue regeneration. AB - One of the most relevant issues in future medicine is tissue regeneration. Transplantation medicine alone cannot solve the problem of incurable conditions of vital organs. One approach to this might be the replication of the spontaneous regeneration that is found in embryonic/neonatal tissue. In this study, a tissue model for basic investigation of regeneration mechanisms in vivo was established. We demonstrated by histology and immunohistochemical staining for types I and II collagen that neonatal rat cartilage unlike adult cartilage has the capacity for rapid scarfree regeneration after full-thickness incision. The underlying mechanism was identified in the preserved proliferative capacity of neonatal chondrocytes. This in vivo model should prove useful in further studies of the role of cellular (e.g., GA cell cycle regulators) and extracellular (e.g., cytokines) factors in tissue regeneration and wound healing. PMID- 11897000 TI - Highlights of the 6th Congress of the International Xenotransplantation Association, Chicago, 2001. PMID- 11897001 TI - The interaction between primate blood and mouse islets induces accelerated clotting with islet destruction. AB - BACKGROUND: Mouse islets transplanted under the renal subcapsular space of cynomolgus monkeys are subject to a form of hyperacute rejection, the mechanism of which is unclear. As islets are in contact with whole blood at the time of transplantation, the effect of platelets and the coagulation cascade on islet destruction was assessed. METHODS: Coagulation was assessed using thromboelastography on citrated/recalcified human blood samples with freshly isolated C57/Bl6 mouse islets. A dynamic islet perifusion system was used to assess the effect of islets on blood cells and coagulation factors. Cytotoxicity was evaluated using (51)Cr labelled islets incubated with human blood and islet destruction was also evaluated using a histological grading system. Continuous PO(2) measurements were made in a static incubation system to assess the role of hypoxia in islet destruction. RESULTS: Mouse islets incubated in human blood induced accelerated coagulation and rapid consumption of platelets within 15 min. Within 1 h of incubation, 52% of mouse islets exposed to xenogeneic human blood showed features of severe damage with necrosis when compared with islets incubated in syngeneic blood. Specific lysis of the xenogeneic islets was demonstrable (Mean percentage lysis: 48%, P < 0.05 vs. control) after 4 h incubation in human blood. Oxygen levels remained constant at a level adequate to maintain islet viability in separate experiments. CONCLUSION: Mouse islets induce rapid activation of the clotting cascade and platelet consumption in vitro when exposed to human blood, which correlated with histological evidence of significant destruction demonstrable within 1 h of exposure to human or non-human primate blood. This in vitro model has features which appear to correlate with the islet destruction seen in vivo and could be a useful model for the study of the mechanisms underlying the rapid destruction of xenogeneic islets in primate recipients. PMID- 11897002 TI - Protection of hDAF-transgenic porcine endothelial cells against activation by human complement: role of the membrane attack complex. AB - Xenograft rejection in the discordant pig-to-primate model is dependent on binding of natural antibodies to gal-alpha [1-3]-gal epitopes on the porcine endothelial cell (EC). This leads to complement activation and deposition of activation products onto the membrane and results in perturbation of EC function and thrombus formation. Here we investigated the ability of human complement activation products to directly induce activation of porcine EC, with subsequent upregulation of adhesion and pro-coagulant molecules. Porcine aortic EC were isolated from wild-type and hDAF-transgenic pigs and incubated with human serum, either in the presence or absence of the soluble complement inhibitor TP10 (sCR1). Recombinant C5a, C1q-IgG immune complexes, C6-deficient human serum and serum containing anti-C9 Ab were used to identify EC activating complement products. Heat-inactivated human serum was used as a negative control. Cells were stained with antibodies against human C3, the MAC or with antibodies cross reactive for porcine E-Selectin, VCAM-1 or Tissue Factor, and analyzed by flow cytometry. We found upregulation of E-Selectin and Tissue Factor on wild-type EC after incubation with human serum. This effect coincided with the deposition of C3 and MAC on the membrane of these cells. The addition of TP10 inhibited EC activation by up to 95%. In contrast, greatly reduced C3 and MAC deposition was detected on hDAF transgenic cells, and no complement-mediated EC activation was seen. Experiments with C6-deficient serum and incubation with anti-C9 Ab indicate a major role of the MAC in serum-mediated EC activation, whereas neither C5a nor C1q-IgG caused activation of EC. These data provide further explanation of the protective role of human DAF in the pig-to-primate xenotransplantation model. PMID- 11897003 TI - Modulation of the in vivo primate anti-Gal response through administration of anti-idiotypic antibodies. AB - Polyclonal anti-idiotypic antibodies (AIA) were generated against human Gal alpha 1,3Gal antibodies (anti-Gal) isolated from a single donor. Specificity of the AIA was demonstrated by selective binding to anti-Gal antibodies (Ab) and absence of reactivity to non-Gal Ab. The idiotopes identified by AIA were present on anti Gal Ab from all of the human samples evaluated (n=59) as well as on pooled samples, demonstrating that a restricted number of dominant idiotopes characterized the human anti-Gal Ab response. Furthermore, the AIA had cross species reactivity with baboon serum samples (n=19), suggesting that the overall shape of the anti-Gal Ab combining site is conserved throughout the Old World primates and providing additional evidence of the limited heterogeneity of the anti-Gal Ab repertoire. In order to evaluate the potential effect of AIA in the modulation of the anti-Gal response in vivo, a baboon was injected with repeated doses of the purified AIA. Following AIA treatment, new Ab were generated that reduced Ab-mediated cytotoxicity to porcine cells. Furthermore, administration of the AIA to a baboon prolonged the survival of intravenously infused pig hematopoietic cells when compared with their survival in a control baboon that did not receive prior AIA treatment but underwent a similar conditioning regimen. PMID- 11897004 TI - Expression of xenogeneic MHC class II molecules in HLA-DR(+) and -DR(-) cells: influence of retrovirus vector design and cellular context. AB - We recently established that molecular chimeras of major histocompatibility complex (MHC) class II molecules, created via retroviral transfer of allogeneic class II cDNAs into bone marrow cells (BMCs), alleviated complications associated with mixed BMC chimeras while leading to T cell tolerance to renal grafts sharing the transferred class II. Initially demonstrated for allogeneic transplants in miniature swine, this concept was extended to T-dependent antibody (Ab) responses to xenogeneic antigens (Ags) in the pig --> baboon combination. Successful down regulation of T cell responses appeared, however, to be contingent on a tight lineage-specific expression of transferred class II molecules. The present studies were, therefore, designed to evaluate the influence of construct design and cellular environment on expression of retrovirally transferred xenogeneic class II cDNAs. Proviral genomes for pig class II SLA-DR expression, differing only at the marker neo(r) or enhanced green fluorescent protein (EGFP) gene, showed increased membrane SLA-DR density on HLA-DR(-) fibroblasts as well as HLA DR(+), TF-1 erythroleukemia cells. More importantly, HLA-DR(+) human B cell lines, although efficiently transduced with pig DR retroviruses, exhibited unstable surface pig DR. Surface pig DR- B cells, nevertheless, stimulated autologous human T cells pre-sensitized to pig Ags, a proliferation likely occurring through presentation of class II-derived peptides. Collectively, these data suggest that surface expression of transferred class II molecules is not related to the ability of recipient cells to synthesize xenogeneic class II molecules but rather to their Ag processing capacities. PMID- 11897005 TI - A soluble chimeric inhibitor of C3 and C5 convertases, complement activation blocker-2, prolongs graft survival in pig-to-rhesus monkey heart transplantation. AB - Complement plays a critical role in many pathologic processes and in xenograft rejection. Therefore, effective complement inhibitors are of great interest. In pig-to-primate organ transplantation, hyperacute rejection results from antibody deposition and complement activation. Complement activation blocker-2 (CAB-2), a recombinant soluble chimeric protein derived from human decay accelerating factor (DAF) and membrane cofactor protein, inhibits C3 and C5 convertases of both classical and alternative pathways. CAB-2 reduces complement-mediated tissue injury of a pig heart perfused ex vivo with human blood. Therefore, we studied the efficacy of CAB-2 when a pig heart is transplanted heterotopically into rhesus monkeys receiving no immunosuppression. Graft survival in three control monkeys was 1.26 +/- 0.2 h; it was markedly prolonged in eight monkeys that received CAB-2. Of the six monkeys that received a single dose of CAB-2 (15 mg/kg i.v.), four had graft survivals of 21, 95, 96, and 108 h, and two died at 7 to 11 h post-transplant with a beating graft, as a result of technical complications. The two monkeys given multiple doses of CAB-2 had graft survivals of 95 and 96 h. CAB-2 markedly inhibited complement activation, as shown by a strong reduction in generation of C3a and SC5b-9. At graft rejection, tissue deposition of iC3b, C4 and C9 was similar or slightly reduced from controls, and deposition of IgG, IgM, C1q and fibrin did not change. Thus, complement inhibition with CAB-2 abrogates hyperacute rejection of pig hearts transplanted into rhesus monkeys, but does not prevent delayed/acute vascular rejection. These studies demonstrate that the beneficial effects of complement inhibition on survival of a pig heart xenograft in rhesus monkeys are similar to those in other primate species and that CAB-2 may be useful in xenotransplantation and other complement-mediated conditions. PMID- 11897006 TI - Anti-pig antibody levels in naive baboons and cynomolgus monkeys. AB - Anti-pig antibodies (APA) were analysed in serum from 28 naive wild-caught baboons (originating from Kenya) and 31 naive captive-bred cynomolgus monkeys (13 from the Philippines and 18 from Mauritius), using a haemolytic assay with pig erythrocytes (APA), flow cytometry on the porcine lymphoma T-cell cell line L35, and enzyme linked immunosorbent assay (ELISA) using alpha-Gal type II and type VI antigen. This was extended in baboon samples by the evaluation in two laboratories (Imutran, Cambridge, UK and Immerge, Boston, USA), and by antibody absorption using either immobilized alpha-Gal type II or alpha-Gal type VI. Anti porcine antibodies were demonstrated in all assays with substantial variability within and between the three non-human primate groups. Immunoglobulin (Ig)M antibody levels tended to be similar to or higher than those in a pooled normal human standard serum while IgG levels tended to be lower. Highest antibody levels were recorded in Mauritius cynomolgus monkeys. There were statistically significant correlations between assays for IgM or IgG class anti-Gal antibodies using either alpha-Gal type II or alpha-Gal type VI as antigen, both for different assays and two laboratories involved. Also, significant correlations were observed between the anti-Gal and L35 binding assays. Baboon sera before and after absorption to immobilized alpha-Gal type II or type VI were analysed for anti-Gal type VI or type II antibody: levels were almost undetectable indicating that most anti-Gal antibodies react to epitopes shared between alpha-Gal type II and type VI oligosaccharides. Finally, the relation between APA and outcome of porcine heart xenotransplantation in cynomolgus monkeys and baboons showed no apparent relation between pre-transplant APA levels and the occurrence of hyperacute rejection (HAR) when compared with non-immunological cause of organ/recipient dysfunction or acute humoral xenograft rejection during the first 4 days post-transplantation or survival exceeding 4 days post-transplantation. PMID- 11897007 TI - Anti-Gal alpha 1-3Gal IgM and IgG antibody levels in sera of humans and old world non-human primates. AB - Organs transplanted from pig to primate are rejected within minutes or hours by an antibody-dependent, complement-mediated mechanism [hyperacute rejection (HAR)]. Even after depletion of anti-Gal alpha 1-3Gal (Gal) antibody (Ab), for example by extracorporeal immunoadsorption, return of natural Ab is believed to be a major factor in the initiation of acute humoral xenograft rejection. Various non-human primates are used as recipients of pig organs in experimental discordant xenotransplantation (XTx) models. However, anti-Gal IgM and IgG levels in non-human primates may differ from those in humans. Serum levels of anti-Gal IgM and IgG were measured by enzyme-linked immunosorbent assay (ELISA) in humans (n=14), chimpanzees (n=8), baboons (n=214), cynomolgus monkeys (n=29), rhesus monkeys (n=23) and Japanese monkeys (n=6). The mean level of anti-Gal IgM was significantly higher in chimpanzees than in other groups, while in rhesus monkeys it was significantly lower than in other groups, except baboons and Japanese monkeys. The mean human anti-Gal IgG level was higher than in other groups and this difference reached statistical significance except with regard to chimpanzees. The mean anti-Gal IgG level in baboons was significantly lower than that in humans, chimpanzees and cynomolgus monkeys. The measured differences in anti-Gal IgM and IgG levels may affect the kinetics of Ab removal and rate of return in different species, and thus may have relevance for translating work in non-human primate models to the clinical setting. PMID- 11897009 TI - Identification and preliminary characterization of mouse Adam33. AB - BACKGROUND: The metalloprotease-disintegrin family, or ADAM, proteins, are implicated in cell-cell interactions, cell fusion, and cell signaling, and are widely distributed among metazoan phyla. Orthologous relationships have been defined for a few ADAM proteins including ADAM10 (Kuzbanian), and ADAM17 (TACE), but evolutionary relationships are not clear for the majority of family members. Human ADAM33 refers to a testis cDNA clone that does not contain a complete open reading frame, but portions of the predicted protein are similar to Xenopus laevis ADAM13. RESULTS: In a 48 kb region of mouse DNA adjacent to the Attractin gene on mouse chromosome 2, we identified sequences very similar to human ADAM33. A full-length mouse cDNA was identified by a combination of gene prediction programs and RT-PCR, and the probable full-length human cDNA was identified by comparison to human genomic sequence in the homologous region on chromosome 20p13. Mouse ADAM33 is 44% identical to Xenopus laevis ADAM13, however a phylogenetic alignment and consideration of functional domains suggests that the two genes are not orthologous. Mouse Adam33 is widely expressed, most highly in the adult brain, heart, kidney, lung and testis. CONCLUSIONS: While mouse ADAM33 is similar to Xenopus ADAM13 in sequence, further examination of its embryonic expression pattern, catalytic activity and protein interactions will be required to assess the functional relationship between these two proteins. Adam33 is expressed in the mouse adult brain and could play a role in complex processes that require cell-cell communication. PMID- 11897010 TI - Cloning and characterization of the mouse Mcoln1 gene reveals an alternatively spliced transcript not seen in humans. AB - BACKGROUND: Mucolipidosis type IV (MLIV) is an autosomal recessive lysosomal storage disorder characterized by severe neurologic and ophthalmologic abnormalities. Recently the MLIV gene, MCOLN1, has been identified as a new member of the transient receptor potential (TRP) cation channel superfamily. Here we report the cloning and characterization of the mouse homologue, Mcoln1, and report a novel splice variant that is not seen in humans. RESULTS: The human and mouse genes display a high degree of synteny. Mcoln1 shows 91% amino acid and 86% nucleotide identity to MCOLN1. Also, Mcoln1 maps to chromosome 8 and contains an open reading frame of 580 amino acids, with a transcript length of approximately 2 kb encoded by 14 exons, similar to its human counterpart. The transcript that results from murine specific alternative splicing encodes a 611 amino acid protein that differs at the c-terminus. CONCLUSIONS: Mcoln1 is highly similar to MCOLN1, especially in the transmembrane domains and ion pore region. Also, the late endosomal/lysosomal targeting signal is conserved, supporting the hypothesis that the protein is localized to these vesicle membranes. To date, there are very few reports describing species-specific splice variants. While identification of Mcoln1 is crucial to the development of mouse models for MLIV, the fact that there are two transcripts in mice suggests an additional or alternate function of the gene that may complicate phenotypic assessment. PMID- 11897011 TI - Locations of several novel 2'-O-methylated nucleotides in human 28S rRNA. AB - BACKGROUND: Ribose 2'-O-methylation, the most common nucleotide modification in mammalian rRNA, is directed by the C/D box small nucleolar RNAs (snoRNAs). Thus far, more than fifty putative human rRNA methylation guide snoRNAs have been identified. For nine of these snoRNAs, the respective ribose methylations in human 28S rRNA have been only presumptive. RESULTS: In this study, the methylation state of human 28S rRNA in the positions predicted by the snoRNAs U21, U26, U31, U48, U50, U73, U74, U80 and U81 was assessed using reverse transcription-based methods and several novel 2'-O-methylations were localized. CONCLUSIONS: Seven novel ribose 2'-O-methylated residues (Am389, Am391, Gm1604, Gm1739, Gm2853, Cm3810, Gm4156, predicted by snoRNAs U26, U81, U80, U73, U50, U74 and U31, respectively) have been localized in human 28S rRNA. The total number of 2'-O-methylations in human rRNA is not yet known. PMID- 11897012 TI - Delta opioid activation of the mitogen-activated protein kinase cascade does not require transphosphorylation of receptor tyrosine kinases. AB - BACKGROUND: In this study, we investigated the mechanism(s) by which delta opioids induce their potent activation of extracellular signal-regulated protein kinases (ERKs) in different cell lines expressing the cloned delta-opioid receptor (delta-OR). While it has been known for some time that OR stimulation leads to the phosphorylation of both ERK isoforms, the exact progression of events has remained elusive. RESULTS: Our results indicate that the transphosphorylation of an endogenous epidermal growth factor receptor (EGFR) in the human embryonic kidney (HEK-293) cell line does not occur when co-expressed delta-ORs are stimulated by the delta-opioid agonist, D-Ser-Leu-enkephalin-Thr (DSLET). Moreover, neither pre-incubation of cultures with the selective EGFR antagonist, AG1478, nor down-regulation of the EGFR to a point where EGF could no longer activate ERKs had an inhibitory effect on ERK activation by DSLET. These results appear to rule out any structural or catalytic role for the EGFR in the delta-opioid-mediated MAPK cascade. To confirm these results, we used C6 glioma cells, a cell line devoid of the EGFR. In delta-OR-expressing C6 glioma cells, opioids produce a robust phosphorylation of ERK 1 and 2, whereas EGF has no stimulatory effect. Furthermore, antagonists to the RTKs that are endogenously expressed in C6 glioma cells (insulin receptor (IR) and platelet-derived growth factor receptor (PDGFR)) were unable to reduce opioid-mediated ERK activation. CONCLUSION: Taken together, these data suggest that the transactivation of resident RTKs does not appear to be required for OR-mediated ERK phosphorylation and that the tyrosine-phosphorylated delta-OR, itself, is likely to act as its own signalling scaffold. PMID- 11897013 TI - Risk of mortality in a cohort of patients newly diagnosed with chronic atrial fibrillation. AB - OBJECTIVE: To estimate the mortality rate of patients newly diagnosed with chronic atrial fibrillation (AF) and compare it with the one in the general population. To evaluate the role of co-morbidity and other factors on the risk of dying among AF patients. METHODS: We used the General Practice Research Database in the UK to perform a retrospective cohort study. We followed a cohort of chronic AF patients (N = 1,035) and an age and sex matched cohort of 5,000 subjects sampled from the general population. We used all deceased AF patients as cases (n = 234) and the remaining AF patients as controls to perform a nested case-control analysis. We estimated mortality risk associated with AF using Cox regression. We computed mortality relative risks using logistic regression among AF patients. RESULTS: During a mean follow-up of two years, 393 patients died in the general population cohort and 234 in the AF cohort. Adjusted relative risk of death in the cohort of AF was 2.5 (95%CI 2.1 - 3.0) compared to the general population. Among AF patients, mortality risk increased remarkably with advancing age. Smokers carried a relative risk of dying close to threefold. Ischaemic heart disease was the strongest clinical predictor of mortality with a RR of 3.0 (95% CI; 2.1-4.1). Current use of calcium channel blockers, warfarin and aspirin was associated with a decreased risk of mortality. CONCLUSIONS: Chronic AF is an important determinant of increased mortality. Major risk factors for mortality in the AF cohort were age, smoking and cardiovascular co-morbidity, in particular ischaemic heart disease. PMID- 11897014 TI - Longitudinal impact of a youth tobacco education program. AB - BACKGROUND: Information on the effectiveness of elementary school level, tobacco use prevention programs is generally limited. This study assessed the impact of a structured, one-time intervention that was designed to modify attitudes and knowledge about tobacco. Participants were fifth-grade students from schools in western New York State. METHODS: Twenty-eight schools, which were in relatively close geographic proximity, were randomized into three groups; Group 1 was used to assess whether attitudes/knowledge were changed in the hypothesized direction by the intervention, and if those changes were retained four months later. Groups 2 and 3, were used as comparison groups to assess possible test-retest bias and historical effects. Groups 1 and 3 were pooled to assess whether attitudes/knowledge were changed by the intervention as measured by an immediate post-test. The non-parametric analytical techniques of Wilcoxon-Matched Pairs/Sign Ranks and the Mann-Whitney-Wilcoxon Rank Sums Tests were used to compare proportions of correct responses at each of the schools. RESULTS: Pooled analyses showed that short-term retention on most items was achieved. It was also found that retention on two knowledge items 'recognition that smokers have yellow teeth and fingers' and 'smoking one pack of cigarettes a day costs several hundred dollars per year' was maintained for four months. CONCLUSIONS: The findings suggest that inexpensive, one-time interventions for tobacco-use prevention can be of value. Changes in attitudes and knowledge conducive to the goal of tobacco-use prevention can be achieved for short-term retention and some relevant knowledge items can be retained for several months. PMID- 11897015 TI - Frailty, fitness and late-life mortality in relation to chronological and biological age. AB - BACKGROUND: People age at remarkably different rates, but how to estimate trajectories of senescence is controversial. METHODS: In a secondary analysis of a representative cohort of Canadians aged 65 and over (n = 2914) we estimated a frailty index based on the proportion of 20 deficits observed in a structured clinical examination. The construct validity of the index was examined through its relationship to chronological age (CA). The criterion validity was examined in its ability to predict mortality, and in relation to other predictions about aging. From the frailty index, relative (to CA) fitness and frailty were estimated, as was an individual's biological age. RESULTS: The average value of the frailty index increased with age in a log-linear relationship (r = 0.91; p < 0.001). In a Cox regression analysis, biological age was significantly more highly associated with death than chronological age. The average increase in the frailty index (i.e. the average accumulation of deficits) amongst those with no cognitive impairment was 3 per cent per year. CONCLUSIONS: The frailty index is a sensitive predictor of survival. As the index includes items not traditionally related to adverse health outcomes, the finding is compatible with a view of frailty as the failure to integrate the complex responses required to maintain function. PMID- 11897016 TI - The father of us all. PMID- 11897017 TI - The gentle art of gene arrangement: the meaning of gene clusters. AB - Genome sequence comparisons reveal that some sets of genes are in similar linkage groups in different organisms while other sets are dispersed. Are some linkage groups maintained by chance, or is there an advantage to such an arrangement? Some insights may come from large clusters of genes, such as the major histocompatibility complex which includes many genes involved in immune defense. PMID- 11897018 TI - The model unicellular eukaryote, Schizosaccharomyces pombe. AB - The fission yeast Schizosaccharomyces pombe has long been a model organism for studies of eukaryotic cells, winning renown especially for studies of the cell cycle. Now that its genome has been sequenced, S. pombe is ready to assume its rightful place in the pantheon of small eukaryotic giants. PMID- 11897020 TI - Using orthologous and paralogous proteins to identify specificity determining residues. AB - BACKGROUND: Concepts of orthology and paralogy are become increasingly important as whole-genome comparison allows their identification in complete genomes. Functional specificity of proteins is assumed to be conserved among orthologs and is different among paralogs. We used this assumption to identify residues which determine specificity of protein-DNA and protein-ligand recognition. Finding such residues is crucial for understanding mechanisms of molecular recognition and for rational protein and drug design. RESULTS: Assuming conservation of specificity among orthologs and different specificity of paralogs, we identify residues which correlate with this grouping by specificity. The method is taking advantage of complete genomes to find multiple orthologs and paralogs. The central part of this method is a procedure to compute statistical significance of the predictions. The procedure is based on a simple statistical model of protein evolution. When applied to a large family of bacterial transcription factors, our method identified 12 residues that are presumed to determine the protein-DNA and protein-ligand recognition specificity. Structural analysis of the proteins and available experimental results strongly support our predictions. Our results suggest new experiments aimed at rational re-design of specificity in bacterial transcription factors by a minimal number of mutations. CONCLUSIONS: While sets of orthologous and paralogous proteins can be easily derived from complete genomic sequences, our method can identify putative specificity determinants in such proteins. PMID- 11897021 TI - A hitchhiker's guide to cell biology: exploitation of host-cell functions by intracellular pathogens. AB - A report on the 'Pathogen-host cell interactions' minisymposium at the 41st Annual Meeting of the American Society for Cell Biology, Washington DC, USA, 8-12 December 2001. PMID- 11897022 TI - Genomics meets nanoscience: probing genes and the cell nucleus at 10-9 meters. AB - A report on the Jackson Laboratory 'Genomics meets nanoscience' meeting, Bar Harbor, USA, 9-12 October 2001. PMID- 11897023 TI - Vector algebra in the analysis of genome-wide expression data. AB - BACKGROUND: Data from thousands of transcription-profiling experiments in organisms ranging from yeast to humans are now publicly available. How best to analyze these data remains an important challenge. A variety of tools have been used for this purpose, including hierarchical clustering, self-organizing maps and principal components analysis. In particular, concepts from vector algebra have proven useful in the study of genome-wide expression data. RESULTS: Here we present a framework based on vector algebra for the analysis of transcription profiles that is geometrically intuitive and computationally efficient. Concepts in vector algebra such as angles, magnitudes, subspaces, singular value decomposition, bases and projections have natural and powerful interpretations in the analysis of microarray data. Angles in particular offer a rigorous method of defining 'similarity' and are useful in evaluating the claims of a microarray based study. We present a sample analysis of cells treated with rapamycin, an immunosuppressant whose effects have been extensively studied with microarrays. In addition, the algebraic concept of a basis for a space affords the opportunity to simplify data analysis and uncover a limited number of expression vectors to span the transcriptional range of cell behavior. CONCLUSIONS: This framework represents a compact, powerful and scalable construction for analysis and computation. As the amount of microarray data in the public domain grows, these vector-based methods are relevant in determining statistical significance. These approaches are also well suited to extract biologically meaningful information in the analysis of signaling networks. PMID- 11897024 TI - Extensive domain shuffling in transcription regulators of DNA viruses and implications for the origin of fungal APSES transcription factors. AB - BACKGROUND: Viral DNA-binding proteins have served as good models to study the biochemistry of transcription regulation and chromatin dynamics. Computational analysis of viral DNA-binding regulatory proteins and identification of their previously undetected homologs encoded by cellular genomes might lead to a better understanding of their function and evolution in both viral and cellular systems. RESULTS: The phyletic range and the conserved DNA-binding domains of the viral regulatory proteins of the poxvirus D6R/N1R and baculoviral Bro protein families have not been previously defined. Using computational analysis, we show that the amino-terminal module of the D6R/N1R proteins defines a novel, conserved DNA binding domain (the KilA-N domain) that is found in a wide range of proteins of large bacterial and eukaryotic DNA viruses. The KilA-N domain is suggested to be homologous to the fungal DNA-binding APSES domain. We provide evidence for the KilA-N and APSES domains sharing a common fold with the nucleic acid-binding modules of the LAGLIDADG nucleases and the amino-terminal domains of the tRNA endonuclease. The amino-terminal module of the Bro proteins is another, distinct DNA-binding domain (the Bro-N domain) that is present in proteins whose domain architectures parallel those of the KilA-N domain-containing proteins. A detailed analysis of the KilA-N and Bro-N domains and the associated domains points to extensive domain shuffling and lineage-specific gene family expansion within DNA virus genomes. CONCLUSIONS: We define a large class of novel viral DNA-binding proteins and their cellular homologs and identify their domain architectures. On the basis of phyletic pattern analysis we present evidence for a probable viral origin of the fungus-specific cell-cycle regulatory transcription factors containing the APSES DNA-binding domain. We also demonstrate the extensive role of lineage-specific gene expansion and domain shuffling, within a limited set of approximately 24 domains, in the generation of the diversity of virus-specific regulatory proteins. PMID- 11897025 TI - Evaluation of thresholds for the detection of binding sites for regulatory proteins in Escherichia coli K12 DNA. AB - BACKGROUND: Sites in DNA that bind regulatory proteins can be detected computationally in various ways. Pattern discovery methods analyze collections of genes suspected to be co-regulated on the evidence, for example, of clustering of transcriptome data. Pattern searching methods use sequences with known binding sites to find other genes regulated by a given protein. Such computational methods are important strategies in the discovery and elaboration of regulatory networks and can provide the experimental biologist with a precise prediction of a binding site or identify a gene as a member of a set of co-regulated genes (a regulon). As more variations on such methods are published, however, thorough evaluation is necessary, as performance may differ depending on the conditions of use. Detailed evaluation also helps to improve and understand the behavior of the different methods and computational strategies. RESULTS: We used a collection of 86 regulons from Escherichia coli as datasets to evaluate two methods for pattern discovery and pattern searching: dyad analysis/dyad sweeping using the program Dyad-analysis, and multiple alignment using the programs Consensus/Patser. Clearly defined statistical parameters are used to evaluate the two methods in different situations. We placed particular emphasis on minimizing the rate of false positives. CONCLUSIONS: As a general rule, sensors obtained from experimentally reported binding sites in DNA frequently locate true sites as the highest-scoring sequences within a given upstream region, especially using Consensus/Patser. Pattern discovery is still an unsolved problem, although in the cases where Dyad-analysis finds significant dyads (around 50%), these frequently correspond to true binding sites. With more robust methods, regulatory predictions could help identify the function of unknown genes. PMID- 11897026 TI - Molecular genetics and structural genomics of the human protein kinase C gene module. AB - BACKGROUND: Protein kinase C (PKC) has become a major focus among cell biologists interested in second-messenger signal transduction and much has been learned about differences in the cellular localization and function of its different isotypes. In this study we systematically address the genomic locations and gene structures of the human PKC gene module. RESULTS: We first carried out fine chromosomal mapping of all nine PKC genes by fluorescence in situ hybridization (FISH), using cosmid and BAC probes. The PKC genes are found to be dispersed throughout the genome, and in some positions distinct from those previously reported: PKCalpha is at 17q24, PKCbeta at 16p12, PKCgamma at 19q13.4, PKCdelta at 3p21.2, PKCepsilon at 2p21, PKCzeta at 1p36.3, PKCeta at 14q22-23, PKCtheta; at 10p15 and PKCiota at 3q26. For PKCiota, an additional FISH signal mapped on Xq21.3 revealed a pseudogene (derived by retrotransposition). PKCgamma, zeta, and theta; are found to map to the most distal positions on the chromosomes, potentially implicating telomere position effects in their expression. Using the complete human genome draft sequence and bioinformatics tools, we then carried out a systematic analysis of PKC gene structure, including determination of the occurrence of single-nucleotide polymorphisms corresponding to the PKC loci. CONCLUSION: This resource of genomic information now facilitates investigation of the PKC gene module in structural chromosomal abnormalities and human disease locus mapping studies. PMID- 11897028 TI - Comparative genomics of Arabidopsis and maize: prospects and limitations. AB - The completed Arabidopsis genome seems to be of limited value as a model for maize genomics. In addition to the expansion of repetitive sequences in maize and the lack of genomic micro-colinearity, maize-specific or highly-diverged proteins contribute to a predicted maize proteome of about 50,000 proteins, twice the size of that of Arabidopsis. PMID- 11897027 TI - Untranslated regions of mRNAs. AB - Gene expression is finely regulated at the post-transcriptional level. Features of the untranslated regions of mRNAs that control their translation, degradation and localization include stem-loop structures, upstream initiation codons and open reading frames, internal ribosome entry sites and various cis-acting elements that are bound by RNA-binding proteins. PMID- 11897029 TI - Integration of splicing, transport and translation to achieve mRNA quality control by the nonsense-mediated decay pathway. AB - When pre-mRNAs are spliced, a multi-component complex is deposited onto them, close to the sites of intron removal. New findings suggest that these exon-exon junction complexes and the complexes that bind mRNA caps are key effectors of the fate of spliced mRNAs and may regulate whether mRNAs containing premature stop codons are degraded. PMID- 11897030 TI - SAGE profiling of the forelimb and hindlimb. AB - A recent study has used serial analysis of gene expression to compare mouse forelimb and hindlimb gene-expression profiles. The method successfully identified known regulators of limb identity and has generated a candidate set of differentially expressed genes that may regulate limb identity. PMID- 11897031 TI - Plant glutathione transferases. AB - SUMMARY: The soluble glutathione transferases (GSTs, EC 2.5.1.18) are encoded by a large and diverse gene family in plants, which can be divided on the basis of sequence identity into the phi, tau, theta, zeta and lambda classes. The theta and zeta GSTs have counterparts in animals but the other classes are plant specific and form the focus of this article. The genome of Arabidopsis thaliana contains 48 GST genes, with the tau and phi classes being the most numerous. The GST proteins have evolved by gene duplication to perform a range of functional roles using the tripeptide glutathione (GSH) as a cosubstrate or coenzyme. GSTs are predominantly expressed in the cytosol, where their GSH-dependent catalytic functions include the conjugation and resulting detoxification of herbicides, the reduction of organic hydroperoxides formed during oxidative stress and the isomerization of maleylacetoacetate to fumarylacetoacetate, a key step in the catabolism of tyrosine. GSTs also have non-catalytic roles, binding flavonoid natural products in the cytosol prior to their deposition in the vacuole. Recent studies have also implicated GSTs as components of ultraviolet-inducible cell signaling pathways and as potential regulators of apoptosis. Although sequence diversification has produced GSTs with multiple functions, the structure of these proteins has been highly conserved. The GSTs thus represent an excellent example of how protein families can diversify to fulfill multiple functions while conserving form and structure. PMID- 11897032 TI - Molecular epidemiology of HIV type 1 infection in Portugal: high prevalence of non-B subtypes. AB - In this study, we have investigated the diversity of current HIV-1 strains circulating in the metropolitan area of Lisbon, Portugal. A total of 217 HIV-1 positive blood samples, collected between October 1998 and December 2000, was genetically characterized in the gp120 C2V3C3 region (n = 205) or part of the gp41 N-terminal segment (n = 12) by heteroduplex mobility assay (HMA) and/or DNA sequencing. The HMA subtyping efficiency (number of samples unambiguously subtyped by HMA divided by the total number of samples subtyped) was 65.9% (143 of 217), with indeterminate migration patterns of subtype A and G strains contributing significantly to this value. On the overall, subtype B was the most prevalent (50.2%), followed by subtypes G (21.7%), A (17.5%), and F (5.5%), whereas subtypes C, D, H, and J accounted altogether for 5.1% of the infections. Non-B subtypes were responsible for 77.4 and 33.1% of the infections among African immigrants and Portuguese subjects, respectively. Angolan individuals (n = 25) were the only ones infected with all the HIV-1 subtypes documented, probably reflecting a high degree of viral genetic diversification in their country of origin. Phylogenetic analysis showed a predominance of IbNG-like viruses among subtype A sequences and two new major subclusters within subtype G (G(P) and G(P)'). The majority of the Portuguese G sequences described formed a well-defined subcluster (G(P)), supported by bootstrap values >90%, phylogenetically distant from clade G sequences in databases. gag (p24/p7) sequence analysis of these variants confirmed the maintenance of the subtype G subclusters. The multiple subclustering observed for the major clades A, B, D, and G, as well as the variety of subtypes found, indicate a high diversity of HIV 1 variants circulating in Portugal and suggest a need for continuous epidemiologic surveillance. PMID- 11897033 TI - Nonadherence to highly active antiretroviral therapy: clinically relevant patient categorization based on electronic event monitoring. AB - Adherence to highly active antiretroviral therapy (HAART) is crucial, but which aspects of drug-taking behavior are important remain largely unknown. In a prospective observational study, 43 HIV-1-infected patients taking HAART underwent electronic event monitoring (EEM). Taking adherence was defined as the percentage of doses taken compared with the number prescribed, dosing adherence was defined as the percentage of days on which all doses were taken, and timing adherence was defined as the percentage of doses taken within 1 hr of the time prescribed. Drug holidays were defined as periods of no drug intake for >24 hr. Cluster analysis, including the four EEM parameters, was used and refined to construct an algorithm to discriminate patients. Patients were categorized as nonadherent if they had a taking adherence of <90%, or a dosing adherence of <75% and at least 1 drug holiday, or a timing adherence of <80% and at least 1 drug holiday, or >6 drug holidays per 100 days. All four EEM parameters differed significantly (p < 0.0001) between the two groups. Adherent patients had a better outcome, as shown by a larger drop in viral load (p = 0.011) and rise in CD4+ cell count (p = 0.035), showing that the algorithm-based categorization is clinically relevant. PMID- 11897034 TI - Insulin-like growth factor I (IGF-I) and IGF-binding protein 3 response to growth hormone is impaired in HIV-infected children. AB - To better characterize the somatotropic axis in HIV-infected children the circadian rhythm of growth hormone (GH), and basal and stimulated (by an insulin like growth factor I [IGF-I] generation test) plasma levels of IGF-I and insulin like growth factor-binding protein 3 (IGFBP-3), were evaluated in 16 children (9 boys and 7 girls; age range, 7-11 years) with HIV infection. All patients were free from active opportunistic infection or liver disease at the time of the study. Sixteen age- and sex-matched healthy children (10 boys and 6 girls; age range, 7-11 years) served as control subjects. GH rhythmometric data were analyzed by single and population mean cosinor analysis. As regards the IGF-I generation test, biosynthetic human GH (hGH, 0.1 IU/kg, 0.033 mg/kg) was administered subcutaneously for 4 days and blood samples were taken from fasting subjects at baseline and on the morning after the last GH injection for measurement of IGF-I and IGFBP-3. Plasma GH levels fell within normal limits in the HIV-seropositive patients and were similar to those of healthy children (1.31 +/- 1.18 vs. 1.57 +/- 1.16 microg/liter, respectively; mean +/- SD). The population mean cosinor analysis shows that the GH circadian rhythm reached statistical significance both in the HIV-seropositive children and in the control group. Despite this, the IGF-I and IGFBP-3 levels were significantly lower in HIV infected children than in the control group (75.6 +/- 57.2 vs. 233.3 +/- 52.5 ng/ml, p < 0.001 and 2.09 +/- 0.17 vs. 3.89 +/- 0.24 mg/liter, p < 0.01, respectively; mean +/- SD); moreover, the response of IGF-I and IGFBP-3 to the IGF-I generation test was significantly lower in HIV-infected children than in the control group (86.3 +/- 55.8 vs. 257.5 +/- 53.4 ng/ml, p < 0.001 and 3.14 +/- 0.43 mg/liter, p < 0.01, respectively; mean +/- SD). It appears that circadian GH secretion is normal in children with HIV infection, but the response to exogenous GH with regard to IGF-I and IGFBP-3 production is impaired, indicating a degree of GH insensitivity in such children. PMID- 11897035 TI - Undefined duration of opiate withdrawal induced by efavirenz in drug users with HIV infection and undergoing chronic methadone treatment. PMID- 11897036 TI - Coreceptor change appears after immune deficiency is established in children infected with different HIV-1 subtypes. AB - Change of HIV-1 coreceptor use has been connected to progression of disease in children infected with HIV-1, presumably subtype B. It has not been possible to discern whether the appearance of new viral phenotypes precedes disease development or comes as a consequence of it. We studied the evolution of coreceptor use in HIV-1 isolates from 24 vertically infected children. Their clinical, virological, and immunological status was recorded and the env V3 subtype was determined by DNA sequencing. Coreceptor use was tested on human cell lines, expressing CD4 together with CCR5, CXCR4, and other chemokine receptors. The children carried five different env subtypes (nine A, five B, four C, three D, and one G) and one circulating recombinant form, CRF01_AE (n = 2). Of the 143 isolates, 86 originated from peripheral blood mononuclear cells (PBMCs) and 57 originated from plasma, received at 90 time points. In 52 of 54 paired plasma and PBMC isolates the coreceptor use was concordant. All 74 isolates obtained at 41 time points during the first year of life used CCR5. A change from use of CCR5 to use of CXCR4 occurred in four children infected with subtype A, D, or CRF01_AE after they had reached 1.5 to 5.8 years of age. There was a significant association with decreased CD4+ cell levels and severity of disease but, interestingly, the coreceptor change appeared months or even years after the beginning of the immunological deterioration. Thus CXCR4-using virus may emerge as a possible consequence of immune deficiency. The results provide new insights into AIDS development in children. PMID- 11897038 TI - HIV type 1-specific IgE in serum of long-term surviving children inhibits HIV type 1 production in vitro. AB - We previously identified a group of long-term pediatric survivors who had acquired HIV-1 through maternal transmission; had not received antiretroviral therapy; are now >8 years old, in good health, and with no opportunistic infections; and have not failed to thrive, although they have greatly decreased numbers of blood CD4+ T cells (<500/mm(3)). All the children have elevated total serum IgE levels (210-2475 IU/ml) and make anti-HIV-1 IgE or IgE directed against non-HIV-1 specificities (radioimmunoassay, Western blot assay); they have no detectable antigenemia. We have now studied the ability of anti-HIV-1 IgE in serum obtained from these children to regulate (1) production of HIV-1 by interleukin 2/phytohemagglutinin (IL-2/PHA)-stimulated peripheral blood mononuclear cells (PBMCs) taken from HIV-1-seronegative donors and infected with a T cell-tropic clone of HIV-1, and (2) transmission of a primary HIV-1 strain from adult AIDS patients to uninfected IL-2/PHA-stimulated PBMCs (p24 core antigen production). High levels of HIV-1 production were observed when PBMCs were cultured for 5 days in the presence of HIV-1-seronegative donor serum that was either IgE positive or IgE negative (IgE, >100 or <100 IU/ml, respectively). HIV-1 production also was observed when PBMCs were cultured with HIV-1-infected donor serum that either contained IgE directed against non-HIV-1 specificities or was IgE negative; these levels were 40% less than those seen with sera from the HIV-1-seronegative donors. Far greater inhibition of virus production was observed if the serum in culture contained anti-HIV-1 IgE (>95%). Virus neutralization did not appear to account for the inhibition obtained with anti HIV-1 IgE-containing serum because virus production was not suppressed in cultures to which serum was added immediately preinfection (<10%), but was strongly suppressed when serum was added 1.5 hr postinfection (>95%). The inhibition of virus production obtained with serum containing anti-HIV-1 IgE was reversed when (1) serum was depleted of IgE (immunoaffinity), but not when it was depleted of IgG (protein G-Sepharose) before inclusion in culture postinfection, (2) anti-IgE, but not anti-IgG, was included in culture, or (3) serum was heat treated before culture. The results indicate that serum from certain HIV-1 infected pediatric long-term survivors contains agents that inhibit HIV-1 production in vitro, and that these agents include anti-HIV-1 IgE. They suggest that a cytotoxic event, rather than virus neutralization, plays an important role in anti-HIV-1 IgE-mediated inhibition of virus production. PMID- 11897037 TI - Increased replication of non-syncytium-inducing HIV type 1 isolates in monocyte derived macrophages is linked to advanced disease in infected children. AB - Non-syncytium-inducing (NSI) strains of HIV-1 prevail among most infected children, including pediatric patients who develop advanced disease, severe immune suppression, and die. A study was designed to address the hypothesis that genotypic and/or phenotypic markers can distinguish NSI viruses isolated during early infection from NSI viruses found in advanced disease. Primary HIV-1 isolates, which were obtained from 43 children, adolescents, and adults who displayed a cross-section of clinical disease and immune suppression but were untreated by protease inhibitor antiretroviral therapy, were characterized for replication phenotype in different cell types. Most individuals (81%) harbored NSI viruses and almost half had progressed to advanced disease or severe immune deficiency. About 51% of NSI isolates produced low levels of p24 antigen (median, 142 pg/ml) in monocyte-derived macrophages (MDMs), 31% produced medium levels (median, 1584 pg/ml), and 17% produced high levels (median, 81,548 pg/ml) (p < 0.001). Seven of eight syncytium-inducing isolates also replicated in MDMs and displayed a dual-tropic phenotype that was associated with advanced disease. Replication of NSI viruses in MDMs varied as much as 100- to 1000-fold and was independent of replication in peripheral blood mononuclear cells. Replication in MDMs provided a clear biological feature to distinguish among viruses that were otherwise identical by NSI phenotype, V3 genotype, and CCR5 coreceptor usage. Low level MDM replication was characteristic of viruses isolated from asymptomatic individuals, including long-term survivors. Enhanced MDM replication was related to morbidity and mortality among patients. Replication levels in MDMs provide a novel prognostic indicator of pathogenic potential by NSI viruses. PMID- 11897039 TI - In vitro replication of SIVcpz is suppressed by beta-chemokines and CD8+ T cells but not by natural killer cells of infected chimpanzees. AB - Unlike humans, chimpanzees are relatively resistant to AIDS after infection with HIV-1 or simian immunodeficiency virus of chimpanzee (SIVcpz). We hypothesized that resistance to disease progression is associated with efficient suppression of virus replication possibly by beta-chemokines secreted by CD8+ lymphocytes and especially natural killer (NK) cells. In vitro suppression of virus replication can be easily studied in SIVcpz-infected chimpanzees because they produce high infectious virus titers in their peripheral blood. A study was undertaken to assess the sensitivity of SIVcpz to beta-chemokines in vitro and to investigate the role of endogenous beta-chemokines in relation to the in vitro capacity of CD8+ lymphocytes and NK cells of chimpanzees to suppress SIVcpz replication. Our results show that SIVcpz uses CCR5 as a coreceptor to gain cell entry and is sensitive to recombinant beta-chemokines in vitro. Here we report that despite their potent capacity to produce RANTES, NK cells of infected chimpanzees do not suppress SIVcpz replication in vitro, in contrast to CD8+ lymphocytes. We also show that endogenous beta-chemokines are not the predominant factors mediating in vitro suppression. PMID- 11897040 TI - Simian AIDS-related lymphoma growth in severe combined immunodeficiency mice is independent of karyotypic abnormalities or Bcl-6 mutations. AB - Simian AIDS-related lymphomas (sARL) of cynomolgus monkeys infected with a simian immunodeficiency virus (SIVsm) were studied in relation to growth in severe combined immunodeficient (SCID) mice, karyotype abnormalities, and DNA sequence of the first noncoding region of the Bcl-6 gene. The tumors were diffuse large B cell lymphomas and expressed a simian homolog to Epstein-Barr virus (HVMF-1) in 12 of 13 primary tumors and corresponding cell lines. A tested cell line was tumorigenic in SCID mice. Tumors in the SCID mice showed cell growth features similar to those in the original lymphoma, suggesting that no subpopulation with growth advantage was selected for in the mice. Spectral karyotype analysis of sARL cell lines showed normal cytogenetic features except for a trisomy of monkey chromosome 2 (corresponding to human chromosomes 7 and 21) in two of five sARL lines, which was not recovered in SCID tumors established from the same cell line. Sequence analysis of a Bcl-6 gene fragment showed sequence variations indicative of population polymorphism(s) in 10 of 13 sARLs, and no evidence of Bcl-6 mutations. Thus Bcl-6 mutations in the first noncoding region are irrelevant for sARL development in cynomolgus monkeys and for tumorigenicity of sARL cell lines. We also demonstrate that no cytogenetic alterations are needed for the development of highly aggressive lymphomas in the SIV-immunosuppressed host. PMID- 11897042 TI - Dietary components and human platelet activity. AB - Platelet hyperactivity is one of the most important factors responsible for the incidence of cardiovascular disease. There are many nutritive and non-nutritive compounds present in the diet which may affect platelet function in various ways. Recent discovery of anti-platelet factors in plants, vegetables and fruits provides a new dietary means for a long-term strategy to favorably modify human blood platelet activity. This review summarises the effects of these dietary components on human platelet function both in vitro and in vivo. PMID- 11897041 TI - Alterations in T lymphocyte profiles of bronchoalveolar lavage fluid from SIV- and Pneumocystis carinii-coinfected rhesus macaques. AB - The goal of this study was to examine SIV- and Pneumocystis carinii-coinfected rhesus macaques as a model of P. carinii infection in HIV-seropositive humans. The influence of P. carinii infection on the cellular composition of bronchoalveolar lavage (BAL) fluid from SIV-infected and normal rhesus macaques was examined by flow cytometric analysis and polymerase chain reaction (PCR). BAL fluid from SIV- and P. carinii coinfected macaques showed a substantial T lymphocyte influx composed of more than 90% CD8+ T cells. These results are in contrast to BAL fluid from SIV-infected macaques with no detectable P. carinii specific PCR product, where CD4+ T cells were present in significant numbers and the CD8+ T cell population was less than 70% of total CD3+ lymphocytes. We observed no significant differences in peripheral blood CD4+ or CD8+ T cell levels in the SIV-infected animals, regardless of P. carinii status, indicating that the CD8+ T cell infiltration in the lungs of the P. carinii-positive animals was likely the result of P. carinii infection. These results demonstrate that although peripheral blood CD4+ T cell levels are predictive of susceptibility to P. carinii infection in this model, the levels are not reflective of the T cell profile in the lung during SIV and P. carinii coinfection. The SIV- and P. carinii-coinfected macaques showed a spectrum of lung disease severity that was histologically similar to human P. carinii pneumonia (PCP). Interestingly, even mild P. carinii infection was sufficient to alter the normal CD4+/CD8+ T cell profiles in the lungs of SIV-infected rhesus macaques. These results are similar to immunologic findings in human AIDS-associated PCP and support the usefulness of this model in the study of immune responses to P. carinii. PMID- 11897043 TI - Platelet adhesion onto a polystyrene surface under static conditions: role of specific platelet receptors and effect of divalent cations. AB - An experimental model was used to elucidate the basic mechanisms involved in the interaction of platelets with an artificial surface. The role of divalent cations and the involvement of specific platelet membrane receptors were evaluated. Isolated platelets were allowed to interact with a polystyrene surface for 20 min in the presence of divalent cations (Ca2+, Mg2+ or Zn2+), a chelating agent (ethylenediaminetetraacetic, EDTA), and specific antibodies to the main platelet receptors, glycoproteins (GP) Ib and IIb-IIIa. The degree of platelet interaction was evaluated using light and electron microscopy. Morphometric analysis was performed to follow up the progression of platelet shape changes after surface activation. Neither Ca2+ nor Mg2+ influenced the number of adherent platelets or the degree of spreading on the polymer. Only Zn2+ induced a statistically significant increase in the rate of platelet adhesion (P<0.01) with higher proportion of fully spread platelets (P<0.01). Chelation of internal pools of divalent cations did not modify the rates of platelet adhesion but prevented platelet spreading. Presence of monoclonal antibodies to GPIb and GP IIb-IIIa did not result in significant differences in the studied parameters. These results suggest that platelet adhesion onto artificial surfaces, in the absence of flow and plasma proteins, is more dependent on cellular motility, where Zn2+ could play an important role, and less dependent on major receptorial mechanisms. PMID- 11897044 TI - Platelet adhesion and aggregation and fibrin formation in flowing blood: a historical contribution by Giulio Bizzozero. AB - At the occasion of the first centennial of Giulio Bizzozero's death, the modern readers' attention is addressed to some ingenious experiments Bizzozero performed in Turin, Italy, around 1880. He discovered and carefully described blood platelet function in flowing conditions and the relationship between platelet adhesion to an artificial surface, aggregation and subsequent fibrin formation and deposition on activated platelet membrane. Bizzozero challenged contemporary concepts involving leukocytes in blood coagulation, but concluded that participation of blood platelets and white cells in fibrin formation was conceivable. PMID- 11897045 TI - Platelet-platelet and platelet-leukocyte interactions induced by outer membrane vesicles from N. meningitidis. AB - A large part of native meningococcal lipopolysaccharide (LPS), i.e., LPS integrated in the outer cell membrane, is released in the form of 'blebs' from surplus outer membrane material. In the present study we investigated the effects of purified outer membrane vesicles (OMVs) on blood platelet-platelet and platelet-leukocyte interactions. Citrated whole blood was stimulated in vitro with equal amounts (on a weight basis) of OMV-integrated LPS, purified LPS (P LPS) from the same meningococcal strain and purified E. coli-LPS. The samples were analyzed by flow cytometry. Upon OMV stimulation platelet aggregation increased 2.1-fold, platelet degranulation 1.8-fold, (measured as CD62P expression), platelet binding to monocytes 2.6-fold, whereas platelet binding to granulocytes increased 2.8-fold. Also, the fraction of large heteroconjugates, i.e., large CD45-positive cell aggregates increased 15.7-fold compared to control. P-LPS and E. coli-LPS also significantly increased platelet aggregation and heteroconjugate formation but did not influence platelet degranulation and binding of platelets to leukocytes in whole blood. When using platelet-rich plasma (PRP), OMVs increased platelet aggregation 2.1-fold and CD62P expression 1.9-fold. P-LPS and E. coli-LPS also significantly increased platelet aggregation in PRP but did not influence platelet degranulation. None of the LPS preparations induced platelet microvesiculation, either in whole blood or in PRP. CONCLUSION: Meningococcal-derived OMVs as well as purified meningococcal LPS, contribute to increased platelet-platelet and platelet-leukocyte aggregation and may thus be of great importance in the development of microthrombosis and organ dysfunction related to fulminant meningococcal septicemia. PMID- 11897046 TI - A novel 196Leu to Pro substitution in the beta3 subunit of the alphaIIbbeta3 integrin in a patient with a variant form of Glanzmann thrombasthenia. AB - Glanzmann thrombasthenia (GT) is an inherited disorder where an absence of platelet aggregation is associated with quantitative or qualitative abnormalities of the alphaIIbbeta3 integrin. In rare patients, amino acid substitutions have provided information on the functional significance of specific domains within alphaIIb or beta3. We now report an elderly male GT patient (R.M.) from the south west of France whose platelets possess a small residual expression of alphaIIbbeta3. Furthermore, the integrin failed to undergo the necessary conformational changes following platelet activation to permit the binding of fibrinogen or activation-dependent monoclonal antibodies despite the presence of an RGD-binding site. Screening of the alphaIIb and beta3 genes by PCR-SSCP revealed a heterozygous mutation at position 685 in exon 5 of the beta3 gene leading to a 196Leu to Pro substitution. 196Leu is a highly conserved amino acid of beta3. The other beta3 allele appeared to be silent. This mutation, inherited from his mother and present in other family members with intermediate levels of alphaIIbbeta3, was close to the MIDAS-like domain of beta3, a fact that appears to explain its effect on alphaIIbbeta3 activation and fibrinogen binding. PMID- 11897047 TI - Altered megakaryocyte-platelet-haemostatic axis in patients with acute stroke. AB - Platelet function is accentuated in acute ischaemic stroke (IS) and may also be altered in haemorrhagic stroke. Whether these changes are a direct reaction to the stroke or are secondary to changes in megakaryocytes (MKs) is unknown. To determine whether MKs are altered in acute stroke we studied 24 patients (18 with ischaemic stroke, six with haemorrhagic stroke) within 3 days of symptom onset, and 14 matched controls. MK ploidy (DNA content, N), size (forward scatter, FSC, arbitrary units), granularity (side scatter, SSC, arbitrary units) and glycoprotein (GP) IIIa expression (arbitrary units) were assessed by flow cytometry. Platelet size (MPV, fl), platelet count (PC, x109/l), circulating reticulated platelets (%), and cutaneous bleeding time (s) were also measured. MK ploidy 22.5 (2.7) vs. 20.6 (1.7) (2p = 0.014); FSC 629 (51) vs. 594 (41) (2p = 0.025); and SSC 843 (88) vs. 776 (76) (2p = 0.020) were each increased, whereas bleeding time 318 (102) vs. 401 (94) (2p = 0.050) was decreased in patients with acute stroke as compared with controls. Trends to increased MK GP IIIa expression and reticulated platelets were also apparent. In a post hoc analysis, the increase in MK ploidy was most prominent in patients with a prior history of hypertension. Ischaemic stroke was associated with non-significant increases in MK ploidy, size, and granularity. However, MK parameters were different in acute haemorrhagic stroke as compared with controls: MK ploidy 23.0 (1.8) vs. 20.6 (1.7) (2p = 0.018); MK FSC 637 (21) vs. 594 (41) (2p = 0.050); MK SSC 872 (41) vs. 776 (76) (2p = 0.020), changes which could be related to the high prevalence of hypertension (83%) in this group. These results demonstrate that pro thrombotic changes in the megakaryocyte-platelet-haemostatic axis (MPHA) are present in acute stroke. Although megakaryocyte changes are likely, in part, to be secondary to the stroke, they could also precede the stroke and therefore explain some of the increased platelet function observed in acute stroke. PMID- 11897048 TI - A variant of the Sebastian platelet syndrome with unique neutrophil inclusions. AB - The Sebastian platelet syndrome (SPS) is a hereditary giant platelet disorder characterized by thrombocytopenia and the presence of neutrophil inclusions identical to those present in neutrophils of patients with another giant platelet disorder, the Fechtner platelet syndrome (FPS). Patients with SPS differ from those with FPS in that they lack the clinical features of the Alport syndrome (high frequency hearing loss, congenital cataracts and chronic interstitial nephritis). The present study has evaluated six patients who resemble individuals with SPS, but have uniquely different neutrophil inclusions. Ultrastructural features of the neutrophil inclusions of the new variant are presented and compared with those found in other giant platelet disorders including classic SPS, FPS and the May-Hegglin anomaly, as well as the Chediak-Higashi syndrome. PMID- 11897050 TI - A comparison of ALPHAScreen, TR-FRET, and TRF as assay methods for FXR nuclear receptors. AB - New developments in detection technologies are providing a variety of biomolecular screening strategies from which to choose. Consequently, we performed a detailed analysis of both separation-based and non-separation-based formats for screening nuclear receptor ligands. In this study, time-resolved fluorescence resonance energy transfer (TR-FRET), ALPHAScreen, and time-resolved fluorescence (TRF) assays were optimized and compared with respect to sensitivity, reproducibility, and miniaturization capability. The results showed that the ALPHAScreen system had the best sensitivity and dynamic range. The TRF assay was more time consuming because of the number of wash steps necessary. The TR-FRET assay had less interwell variation, most likely because of ratiometric measurement. Both the ALPHAScreen and the TR-FRET assays were miniaturized to 8 microl volumes. Of the photomultiplier tube-based readers, the ALPHAScreen reader (ALPHAQuest) presented the advantage of faster reading times through simultaneous reading with four photomultiplier tubes. PMID- 11897051 TI - Automated high throughput screening for serine kinase inhibitors using a LEADseeker scintillation proximity assay in the 1536-well format. AB - High-throughput screening in the 1536-well format has been largely restricted to solution-based and cell-based screens. In this article, we show the feasibility of a completely automated, robust scintillation proximity assay in the 1536-well format that is suitable to identify inhibitors for a serine/threonine kinase from a compound library. The introduction of [(33)P]phosphate into a biotinylated peptide substrate mirrors the activity of the kinase. The peptide is immobilized on streptavidin-coated LEADseeker imaging beads and [(33)P]phosphate incorporation is detected with the LEADseeker imaging system of Amersham Pharmacia Biotech. To improve the liquid handling procedures for imaging bead suspensions in the low microliter range, we developed a novel trough with an integrated stirring function. A comparison of the 1536-well assay to a 384-well assay revealed a comparable assay quality with Z' factors of about 0.7 for the 384-well format and 0.6 for the 1536-well format. In an automated screen of a random compound collection, 94.4% of the inhibitory compounds could be identified with both assay formats. Dose-response curves were performed for a selection of identified kinase inhibitors and revealed similar IC(50) values for both assay formats. PMID- 11897052 TI - Determination of ligand-MurB interactions by isothermal denaturation: application as a secondary assay to complement high throughput screening. AB - We used a temperature-jump isothermal denaturation procedure with various methods of detection to evaluate the quality of putative inhibitors of MurB discovered by high-throughput screening. Three optical methods of detection-ultraviolet hyperchromicity of absorbance, fluorescence of bound dyes, and circular dichroism as well as differential scanning calorimetry were used to dissect the effects of two chemical compounds and a natural substrate on the enzyme. The kinetics of the denaturation process and binding of the compounds detected by quenching of flavin fluorescence were used to quantitate the dose dependencies of the ligand effects. We found that the first step in the denaturation of MurB is the rapid loss of flavin from the active site and that the two chemical inhibitors appeared to destabilize the interaction of the cofactor with the enzyme but stabilize the global unfolding. The kinetics of the denaturation process as well as the loss of flavin fluorescence on binding established that both compounds had nanomolar affinities for the enzyme. We showed that coupling of the various detection methods with isothermal denaturation yields a powerful regimen to provide analytical data for assessing inhibitor specificity for a protein target. PMID- 11897054 TI - A homogeneous high throughput nonradioactive method for measurement of functional activity of Gs-coupled receptors in membranes. AB - A method is described for measuring the activity of G(s)-coupled receptors in a nonradioactive homogeneous membrane-based assay. This method has several major advantages over currently used methods for measuring functional activity of G(s) coupled receptors. The assay is high throughput (>150,000 data points/day using a single reader). Dimethyl sulfoxide tolerance is high ( approximately 10%). Compared to complex cell-based assays, there is limited potential for nonspecific compound action. This resulted in low compound hit rates in robustness screening, where hit rates from a simulated screen were 1.0% (antagonist screen) and 0.1% (agonist screen). No continuous cell culture is required for the assay, reducing cell culture overheads and allowing the screen to run every day. Automation is simple and requires no temperature- or humidity-controlled incubation. No radioactivity is required. The method relies on measurement of cyclic AMP (cAMP) generation by fluorescence polarization assay using commercially available reagents. Membranes (1-2 microg protein per well, containing anti-cAMP antibody) are transferred to 384-well plates containing 1 microl test compound. For antagonist screens, agonist is added 15 min later. After 30 min incubation at room temperature, one further assay reagent (fluorescein-cAMP in a buffer containing detergent) is added. The signal may be read after 1 h and is stable for greater than 12 h. Typical Z' for the assay is approximately 0.5. PMID- 11897053 TI - Microplate screening of the differential effects of test agents on Hoechst 33342, rhodamine 123, and rhodamine 6G accumulation in breast cancer cells that overexpress P-glycoprotein. AB - A microplate screening method has been developed to evaluate the effects of test agents on the accumulation of the fluorescent P-glycoprotein (Pgp) substrates Hoechst 33342, rhodamine 123, and rhodamine 6G in multidrug-resistant (MDR) breast cancer cells that overexpress Pgp. All three substrates exhibit substantially higher accumulation in MCF7 non-MDR cells versus NCI/ADR-RES MDR cells, while incubation with 50 microM reserpine significantly reduces or eliminates these differences. Rhodamine 123 shows the lowest substrate accumulation efficiency in non-MDR cells relative to the substrate incubation level. The effects of several chemosensitizing agents and a series of paclitaxel analogs on the accumulation of each fluorescent substrate suggest that there are distinct differences in the substrate interaction profiles exhibited by these different agents. The described methods may be useful in Pgp-related research in the areas of cancer MDR, oral drug absorption, the blood-brain barrier, renal/hepatic transport processes, and drug-drug interactions. PMID- 11897055 TI - Binding of a Pleckstrin homology domain protein to phosphoinositide in membranes: a miniaturized FRET-based assay for drug screening. AB - Pleckstrin homology (PH) domains are present in key proteins involved in many vital cell processes. For example, the PH domain of Bruton's tyrosine kinase (Btk) binds to phosphatidylinositol triphosphate (PIP(3)) in the plasma membrane after stimulation of the B-cell receptor in B cells. Mutations in the Btk PH domain result in changes in its affinity for PIP(3), with higher binding leading to cell transformation in vitro and lower binding leading to antibody deficiencies in both humans and mice. We describe here a fluorescence resonance energy transfer (FRET)-based biochemical assay that directly monitors the interaction of a PH domain with PIP(3) at a membrane surface. We overexpressed a fusion protein consisting of an enhanced green fluorescent protein (GFP) and the N-terminal 170 amino acids of a Tec family kinase that contains its PH domain (PH170). Homogeneous unilamellar vesicles were made that contained PIP(3) and octadecylrhodamine (OR), a lipophilic FRET acceptor for GFP. After optimization of both protein and vesicle components, we found that binding of the GFP-PH170 protein to PIP3 in vesicles that contain OR results in about a 90% reduction of GFP fluorescence. Using this assay to screen 1440 compounds, we identified three that efficiently inhibited binding of GFP-PH170 to PIP(3) in vesicles. This biochemical assay readily miniaturized to 1.8-microl reaction volumes and was validated in a 3456-well screening format. PMID- 11897056 TI - Adaptation of aequorin functional assay to high throughput screening. AB - AequoScreen, a cellular aequorin-based functional assay, has been optimized for luminescent high-throughput screening (HTS) of G protein-coupled receptor (GPCRs). AequoScreen is a homogeneous assay in which the cells are loaded with the apoaequorin cofactor coelenterazine, diluted in assay buffer, and injected into plates containing the samples to be tested. A flash of light is emitted following the calcium increase resulting from the activation of the GPCR by the sample. Here we have validated a new plate reader, the Hamamatsu Photonics FDSS6000, for HTS in 96- and 384-well plates with CHO-K1 cells stably coexpressing mitochondrial apoaequorin and different GPCRs (AequoScreen cell lines). The acquisition time, plate type, and cell number per well have been optimized to obtain concentration-response curves with 4000 cells/well in 384 well plates and a high signal:background ratio. The FDSS6000 and AequoScreen cell lines allow reading of twenty 96- or 384-well plates in 1 h with Z' values of 0.71 and 0.78, respectively. These results bring new insights to functional assays, and therefore reinforce the interest in aequorin-based assays in a HTS environment. PMID- 11897057 TI - Comparison of high throughput screening technologies for luminescence cell-based reporter screens. AB - As higher density formats become more and more common in HTS labs, the expectations for maintaining faster, lower cost screens puts great pressure on traditional 96-well screens. In some cases higher density formats are not compatible with the assay. This seems especially true in cell-based assays. In our case, the nature of the cells' response forced us to remain in 96-well plates. In this paper, we describe the development of a luminescence reporter assay and its performance in two detection modes, flash and glow. The advantages in cost and throughput for each technique are explored, along with automation considerations. An additional new technology, the use of pins for low-volume transfers, is also briefly described because of its dramatic effect on our screen's throughput. However, it will be more thoroughly presented in a future publication. Comparing the technologies available for HTS aids in designing automated systems that meet the unique needs of each assay. PMID- 11897059 TI - An effective tool for homogeneous assays using a simple robot arm with Stackers TRISTAN. AB - The Twister & RapidPlate Integrated System by TANabe, or TRISTAN, consists of a 96-channel dispenser (RapidPlate 96), a plate reader (V-MAX), and a simple robot arm (Twister). We developed TRISTAN for effectively conducting a homogeneous assay. Although this system accommodates fewer than 20 microplates, it has several advantages over conventional robotic systems for high-throughput screening in the following aspects: parameter setting, running time, hardware errors, manpower, and cost-effectiveness. The system proved to be effective and efficient for homogeneous assays. PMID- 11897058 TI - A novel membrane potential-sensitive fluorescent dye improves cell-based assays for ion channels. AB - The study of ion channel-mediated changes in membrane potential using the conventional bisoxonol fluorescent dye DiBAC(4)(3) has several limitations, including a slow onset of response and multistep preparation, that limit both the fidelity of the results and the throughput of membrane potential assays. Here, we report the characterization of the FLIPR Membrane Potential Assay Kit (FMP) in cells expressing voltage- and ligand-gated ion channels. The steady-state and kinetics fluorescence properties of FMP were compared with those of DiBAC(4)(3), using both FLIPR and whole-cell patch-clamp recording. Our experiments with the voltage-gated K(+) channel, hElk-1, revealed that FMP was 14-fold faster than DiBAC(4)(3) in response to depolarization. On addition of 60 mM KCl, the kinetics of fluorescence changes of FMP using FLIPR were identical to those observed in the electrophysiological studies using whole-cell current clamp. In addition, KCl concentration-dependent increases in FMP fluorescence correlated with the changes of membrane potential recorded in whole-cell patch clamp. In studies examining vanilloid receptor-1, a ligand-gated nonselective cation channel, FMP was superior to DiBAC(4)(3) with respect to both kinetics and amplitude of capsaicin induced fluorescence changes. FMP has also been used to measure the activation of K(ATP) and hERG. Thus this novel membrane potential dye represents a powerful tool for developing high-throughput screening assays for ion channels. PMID- 11897061 TI - Vector interactions and molecular adaptations of lyme disease and relapsing fever spirochetes associated with transmission by ticks. AB - Pathogenic spirochetes in the genus Borrelia are transmitted primarily by two families of ticks. The Lyme disease spirochete, Borrelia burgdorferi, is transmitted by the slow-feeding ixodid tick Ixodes scapularis, whereas the relapsing fever spirochete, B. hermsii, is transmitted by Ornithodoros hermsi, a fast-feeding argasid tick. Lyme disease spirochetes are generally restricted to the midgut in unfed I. scapularis. When nymphal ticks feed, the bacteria pass through the hemocoel to the salivary glands and are transmitted to a new host in the saliva after 2 days. Relapsing fever spirochetes infect the midgut in unfed O. hermsi but persist in other sites including the salivary glands. Thus, relapsing fever spirochetes are efficiently transmitted in saliva by these fast feeding ticks within minutes of their attachment to a mammalian host. We describe how B. burgdorferi and B. hermsii change their outer surface during their alternating infections in ticks and mammals, which in turn suggests biological functions for a few surface-exposed lipoproteins. PMID- 11897062 TI - Traditional and molecular techniques for the study of emerging bacterial diseases: one laboratory's perspective. AB - Identification of emerging bacterial pathogens generally results from a chain of events involving microscopy, serology, molecular tools, and culture. Because of the spectacular molecular techniques developed in the last decades, some authors think that these techniques will shortly supplant culture. The key steps that led to the discovery of emerging bacteria have been reviewed to determine the real contribution of each technique. Historically, microscopy has played a major role. Serology provided indirect evidence for causality. Isolation and culture were crucial, as all emerging bacteria have been grown on artificial media or cell lines or at least propagated in animals. With the use of broad-range polymerase chain reaction, some bacteria have been identified or detected in new clinical syndromes. Culture has irreplaceable advantages for studying emerging bacterial diseases, as it allows antigenic studies, antibiotic susceptibility testing, experimental models, and genetic studies to be carried out, and remains the ultimate goal of pathogen identification. PMID- 11897063 TI - Current status of antimicrobial resistance in Taiwan. AB - While some trends in antimicrobial resistance rates are universal, others appear to be unique for specific regions. In Taiwan, the strikingly high prevalence of resistance to macrolides and streptogramin in clinical isolates of gram-positive bacteria correlates with the widespread use of these agents in the medical and farming communities, respectively. The relatively low rate of enterococci that are resistant to glycopeptide does not parallel the high use of glycopeptides and extended-spectrum beta-lactams in hospitals. The evolving problem of extended spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates is substantial, and some unique enzymes have been found. Recently, some gram-negative bacteria (e.g., Pseudomonas aeruginosa and Acinetobacter baumannii) that are resistant to all available antimicrobial agents including carbapenems have emerged. PMID- 11897064 TI - An outbreak of Rift Valley fever in Northeastern Kenya, 1997-98. AB - In December 1997, 170 hemorrhagic fever-associated deaths were reported in Garissa District, Kenya. Laboratory testing identified evidence of acute Rift Valley fever virus (RVFV). Of the 171 persons enrolled in a cross-sectional study, 31(18%) were anti-RVFV immunoglobulin (Ig) M positive. An age-adjusted IgM antibody prevalence of 14% was estimated for the district. We estimate approximately 27,500 infections occurred in Garissa District, making this the largest recorded outbreak of RVFV in East Africa. In multivariable analysis, contact with sheep body fluids and sheltering livestock in one s home were significantly associated with infection. Direct contact with animals, particularly contact with sheep body fluids, was the most important modifiable risk factor for RVFV infection. Public education during epizootics may reduce human illness and deaths associated with future outbreaks. PMID- 11897065 TI - Surveillance for unexplained deaths and critical illnesses due to possibly infectious causes, United States, 1995-1998. AB - Population-based surveillance for unexplained death and critical illness possibly due to infectious causes (UNEX) was conducted in four U.S. Emerging Infections Program sites (population 7.7 million) from May 1, 1995, to December 31, 1998, to define the incidence, epidemiologic features, and etiology of this syndrome. A case was defined as death or critical illness in a hospitalized, previously healthy person, 1 to 49 years of age, with infection hallmarks but no cause identified after routine testing. A total of 137 cases were identified (incidence rate 0.5 per 100,000 per year). Patients' median age was 20 years, 72 (53%) were female, 112 (82%) were white, and 41 (30%) died. The most common clinical presentations were neurologic (29%), respiratory (27%), and cardiac (21%). Infectious causes were identified for 34 cases (28% of the 122 cases with clinical specimens); 23 (68%) were diagnosed by reference serologic tests, and 11 (32%) by polymerase chain reaction-based methods. The UNEX network model would improve U.S. diagnostic capacities and preparedness for emerging infections. PMID- 11897066 TI - Lack of evidence for human-to-human transmission of avian influenza A (H9N2) viruses in Hong Kong, China 1999. AB - In April 1999, isolation of avian influenza A (H9N2) viruses from humans was confirmed for the first time. H9N2 viruses were isolated from nasopharyngeal aspirate specimens collected from two children who were hospitalized with uncomplicated, febrile, upper respiratory tract illnesses in Hong Kong during March 1999. Novel influenza viruses have the potential to initiate global pandemics if they are sufficiently transmissible among humans. We conducted four retrospective cohort studies of persons exposed to these two H9N2 patients to assess whether human-to-human transmission of avian H9N2 viruses had occurred. No serologic evidence of H9N2 infection was found in family members or health-care workers who had close contact with the H9N2-infected children, suggesting that these H9N2 viruses were not easily transmitted from person to person. PMID- 11897068 TI - Buruli ulcer in Ghana: results of a national case search. AB - A national search for cases of Buruli ulcer in Ghana identified 5,619 patients, with 6,332 clinical lesions at various stages. The overall crude national prevalence rate of active lesions was 20.7 per 100,000, but the rate was 150.8 per 100,000 in the most disease-endemic district. The case search demonstrated widespread disease and gross underreporting compared with the routine reporting system. The epidemiologic information gathered will contribute to the design of control programs for Buruli ulcer. PMID- 11897067 TI - Community-acquired Klebsiella pneumoniae bacteremia: global differences in clinical patterns. AB - We initiated a worldwide collaborative study, including 455 episodes of bacteremia, to elucidate the clinical patterns of Klebsiella pneumoniae. Historically, community-acquired pneumonia has been consistently associated with K. pneumoniae. Only four cases of community-acquired bacteremic K. pneumoniae pneumonia were seen in the 2-year study period in the United States, Argentina, Europe, or Australia; none were in alcoholics. In contrast, 53 cases of bacteremic K. pneumoniae pneumonia were observed in South Africa and Taiwan, where an association with alcoholism persisted (p=0.007). Twenty-five cases of a distinctive syndrome consisting of K. pneumoniae bacteremia in conjunction with community-acquired liver abscess, meningitis, or endophthalmitis were observed. A distinctive form of K. pneumoniae infection, often causing liver abscess, was identified, almost exclusively in Taiwan. PMID- 11897069 TI - Clinical significance and epidemiology of NO-1, an unusual bacterium associated with dog and cat bites. AB - From 1974 to 1998, 22 isolates of an unusual bacterium, designated as CDC nonoxidizer 1 group (NO-1), were sent to the Centers for Disease Control and Prevention for identification. The organism's phenotypic characteristics were similar to asaccharolytic strains of Acinetobacter, but differed in their cellular morphology and cellular fatty acid profile. We report here on NO-1's clinical and epidemiologic significance. In all cases, isolates were recovered from an animal bite wound; 17 (77%) were isolated from a dog bite wound, 4 (18%) from a cat bite wound, and one (5%) from an unspecified animal bite. Clinical data were retrieved and reviewed for 12 (55%) of the 22 bite victims. None of the patients had pre-existing conditions associated with immunosuppression. Seven (58%) patients were hospitalized for a median stay of 4 days (range, 2 to 11 days). The median time between bite to the worsening of symptoms was 17.5 hours (range, 3 to 78 hours). All patients recovered following antibiotic treatment. PMID- 11897070 TI - Comparative antibiotic resistance of diarrheal pathogens from Vietnam and Thailand, 1996-1999. AB - Antimicrobial resistance rates for shigella, campylobacter, nontyphoidal salmonella, and enterotoxigenic Escherichia coli were compared for Vietnam and Thailand from 1996 to 1999. Resistance to trimethoprim-sulfamethoxazole, ampicillin, chloramphenicol, and tetracycline was common. Quinolone resistance remains low in both countries, except among campylobacter and salmonella organisms in Thailand. Nalidixic acid resistance among salmonellae has more than doubled since 1995 (to 21%) in Thailand but is not yet documented in Vietnam. Resistance to quinolones correlated with resistance to azithromycin in both campylobacter and salmonella in Thailand. This report describes the first identification of this correlation and its epidemiologic importance among clinical isolates. These data illustrate the growing magnitude of antibiotic resistance and important differences between countries in Southeast Asia. PMID- 11897071 TI - Epidemiology of Burkholderia cepacia complex in patients with cystic fibrosis, Canada. AB - The Burkholderia cepacia complex is an important group of pathogens in patients with cystic fibrosis (CF). Although evidence for patient-to-patient spread is clear, microbial factors facilitating transmission are poorly understood. To identify microbial clones with enhanced transmissibility, we evaluated B. cepacia complex isolates from patients with CF from throughout Canada. A total of 905 isolates from the B. cepacia complex were recovered from 447 patients in 8 of the 10 provinces; 369 (83%) of these patients had genomovar III and 43 (9.6%) had B. multivorans (genomovar II). Infection prevalence differed substantially by region (22% of patients in Ontario vs. 5% in Quebec). Results of typing by random amplified polymorphic DNA analysis or pulsed-field gel electrophoresis indicated that strains of B. cepacia complex from genomovar III are the most potentially transmissible and that the B. cepacia epidemic strain marker is a robust marker for transmissibility. PMID- 11897072 TI - Broad-range bacterial detection and the analysis of unexplained death and critical illness. AB - Broad-range rDNA polymerase chain reaction (PCR) provides an alternative, cultivation-independent approach for identifying pathogens. In 1995, the Centers for Disease Control and Prevention initiated population-based surveillance for unexplained life-threatening infections (Unexplained Death and Critical Illness Project [UNEX]). To address the causes of UNEX cases, we examined 59 specimens from 46 cases by using broad-range bacterial 16S rDNA PCR and phylogenetic analysis of amplified sequences. Specimens from eight cases yielded sequences from Neisseria meningitidis (cerebrospinal fluid from two patients with meningitis), Streptococcus pneumoniae (cerebrospinal fluid from one patient with meningitis2 and pleural fluid from two patients with pneumonia), or Stenotrophomonas maltophilia (bone marrow aspirate from one patient with pneumonia). Streptococcus pneumoniae rDNA sequence microheterogeneity was found in one pleural fluid specimen, suggesting the presence of multiple strains. In conclusion, known bacterial pathogens cause some critical illnesses and deaths that fail to be explained with traditional diagnostic methods. PMID- 11897073 TI - Antibiotic resistance patterns of bacterial isolates from blood in San Francisco County, California, 1996-1999. AB - Countywide antibiotic resistance patterns may provide additional information from that obtained from national sampling or individual hospitals. We reviewed susceptibility patterns of selected bacterial strains isolated from blood in San Francisco County from January 1996 to March 1999. We found substantial hospital to-hospital variability in proportional resistance to antibiotics in multiple organisms. This variability was not correlated with hospital indices such as number of intensive care unit or total beds, annual admissions, or average length of stay. We also found a significant increase in methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus, and proportional resistance to multiple antipseudomonal antibiotics. We describe the utility, difficulties, and limitations of countywide surveillance. PMID- 11897074 TI - Prosthetic valve endocarditis caused by Bartonella quintana. AB - We describe the first case of Bartonella quintana endocarditis affecting a prosthetic valve in a person with no known risk factors for this infection. Bartonella should be considered as a cause of endocarditis in any clinical setting. PMID- 11897075 TI - Cryptosporidium muris infection in an HIV-infected adult, Kenya. AB - We describe a case of Cryptosporidium muris infection in an HIV-infected adult with diarrhea in Kenya. Sequence analysis of an 840-bp region of the 18S rRNA gene locus demonstrated the isolate had 100% nucleotide identity with C. muris recovered from a rock hyrax, 98.8% with a C. muris "calf" isolate, 95.5% with C. serpentis, but only 87.8% with C. parvum "human" type. PMID- 11897076 TI - Rickettsia felis infection acquired in Europe and documented by polymerase chain reaction. AB - We report the first case of Rickettsia felis infection in Europe to be documented by polymerase chain reaction (PCR) and serologic testing. PMID- 11897077 TI - Hemorrhagic fever with renal syndrome presenting with hemophagocytic lymphohistiocytosis. AB - Hemophagocytic lymphohistiocytosis--which is associated with a variety of infections, malignant neoplasms, autoimmune diseases, and immunodeficiencies--is an uncommon syndrome with a rapidly fatal outcome. We describe the first case of hemorrhagic fever with renal syndrome due to Hantaan virus presenting with reactive hemophagocytosis. PMID- 11897078 TI - Distemper outbreak and its effect on African wild dog conservation. AB - In December 2000, an infectious disease spread through a captive breeding group of African wild dogs (Lycaon pictus) in Tanzania, killing 49 of 52 animals within 2 months. The causative agent was identified as Canine distemper virus (CDV) by means of histologic examination, virus isolation, reverse transcriptase polymerase chain reaction analysis, and nucleotide sequencing. This report emphasizes the importance of adequate protection against infectious diseases for the successful outcome of captive breeding programs of endangered species. PMID- 11897079 TI - Postexposure treatment and animal rabies, Ontario, 1958-2000. AB - This paper investigates the relationship between animal rabies and postexposure treatment (PET) in Ontario by examining the introduction of human diploid cell vaccine (HDCV) in 1980 and the initiation of an oral rabies vaccination program for wildlife in 1989. Introducing HDCV led to an immediate doubling of treatments. Both animal rabies and human treatments declined rapidly after the vaccination program was introduced, but human treatments have leveled off at approximately 1,000 per year. PMID- 11897080 TI - Multidrug-resistant Pseudomonas aeruginosa bloodstream infections: analysis of trends in prevalence and epidemiology. PMID- 11897081 TI - Clinical issues in the prophylaxis, diagnosis, and treatment of anthrax. PMID- 11897082 TI - Public health assessment of potential biological terrorism agents. PMID- 11897083 TI - Interaction between brain L-type calcium channels and alpha2-adrenoceptors in the inhibition of sodium appetite. AB - Calcium channels mediate the actions of many drugs. The present work investigated whether diltiazem, an L-type calcium channel blocker, alters the inhibition of sodium appetite induced by noradrenaline and the alpha2-adrenoceptor agonist clonidine. Adult male Holtzman rats (N=4-8) with cannula implanted into the third cerebral ventricle were submitted to sodium depletion (furosemide sc+24-h removal of ambiente sodium). Sodium depleted control animals that received 0.9% NaCl as vehicle injected intracerebroventricularly (i.c.v.) ingested 13.0+/-1.5 ml/120 min of 1.8% NaCl. Intracerebroventricular injection of either noradrenaline (80 nmol) or clonidine (20 nmol) inhibited 1.8% NaCl intake from 70 to 90%. Prior i.c.v. injection of diltiazem (6-48 nmol) inhibited from 50 to 100% the effect of noradrenaline and clonidine in a dose-response manner. Diltiazem alone at 100 nmol inhibited, but at 50 nmol had no effect on, sodium appetite. The results suggest: (1) common ionic mechanisms involving calcium channels for the inhibition that noradrenaline and clonidine exert on sodium appetite and (2) a dual role for the benzothiazepine site of L-type calcium channels in the control of sodium appetite. PMID- 11897084 TI - Evaluation of 3-nitrotyrosine as a marker for 3-nitropropionic acid-induced oxidative stress in Lewis and Wistar rats and strain-specific whole brain spheroid cultures. AB - The present study investigated whether 3-nitrotyrosine is an early marker for neurodegenerative processes involving oxidative stress. We characterized the 3 nitrotyrosine formation after 3-nitropropionic acid (3-NP) exposure in the whole brain spheroid culture model and in a rat model, using Lewis and Wistar rats. Increased 3-nitrotyrosine concentration in spheroid cultures from Lewis rats was observed at lower dose of and shorter exposure time to 3-NP as compared to alterations in glial fibrillary acidic protein concentration, decrease in glutamine synthetase activity or cell loss. Five days of exposure to 3-NP (5 mM) resulted in decreased staining of GABAergic processes, while neuronal nitric oxide synthase staining was preserved. In addition, staining of EAAC1, anti-2',3' cyclic nucleotide 3'-phosphohydrolase and ED1 was diminished after treatment of spheroid cultures with 3-nitropropionic acid (5 mM), while isolectin B4 staining was increased. Dithiothreitol and vitamin E inhibited the increased formation of 3-nitrotyrosine. Interestingly, N(G)-nitro-L-arginine methyl ester increased the 3-nitrotyrosine formation. No increased 3-nitrotyrosine concentration was shown after exposure to 3-nitropropionic acid during 5 days in spheroid cultures obtained from Wistar rats. In the striatum of 3-NP-exposed Lewis and Wistar rats, no change in 3-nitrotyrosine concentration was observed, whereas only in Wistar rats the glial fibrillary acidic protein concentration was increased in addition to activation of microglial cells. It is concluded that 3-nitrotyrosine was a more sensitive marker for oxidative stress-induced neurodegeneration than glial fibrillary acidic protein and glutamine synthase in spheroid cell cultures of Lewis rats. Finally, the similarities between the 3-NP spheroid model and the vivo model indicate that the spheroid cultures provide a good alternative for chronic exposure of animals to neurotoxins. PMID- 11897085 TI - Sculpting the visual map: the distribution and function of serotonin-1A and serotonin-1B receptors in the optic tectum of the frog. AB - Agonists of serotonin (5-HT)-1 receptors modulate the synaptic strength of the connection between retinal ganglion cells and neurons of the frog optic tectum in brain slices (Brain Res. 1998;781:167-181). We have now used autoradiographic receptor binding techniques to determine the location of 5-HT1A and 5-HT1B binding sites in the laminated optic tectum. 5-HT1A binding sites, as labeled with [3H]8-hydroxy-dipropylaminotetralin (8-OH-DPAT), were highest in the superficial, retinorecipient layers of the tectum, intermediate in layers 6 and 7 and low in the remaining layers. Binding densities in all of these layers were unaffected by optic nerve lesion. 5-HT1B binding sites were visualized using [125I]iodocyanopindolol (ICYP). Binding densities were highest in the plexiform layers 5 and 7 and intermediate in layers 6 and 8. Binding sites were present at low levels in layer 9; however, optic nerve lesion resulted in a strong upregulation of these sites in this layer. Pharmacological manipulation of receptor activation resulted in changes in the activity-dependent visual map that is created at the tectum by retinal ganglion cell terminals. Chronic treatment of the tectum with SB-224289, a selective antagonist of 5-HT1B receptors, disrupted the topographic map. In contrast, exposure to WAY-100635, a selective antagonist of 5-HT1A receptors, refined it. We conclude that both 5-HT1A and 5-HT1B receptors are present in the adult frog tectum and that changes in their activation levels can produce changes in retinotectal transmission levels that drive visual plasticity in opposite directions. PMID- 11897086 TI - Caloric restricted male rats demonstrate fewer synapses in layer 2 of sensorimotor cortex. AB - Previous studies have demonstrated an age-related decline in the density of presumptive inhibitory synapses in layer 2 of rat sensorimotor cortex [J. Comp. Neurol. 439(1) (2001) 65]. Caloric restriction has been shown to ameliorate age related deterioration in a variety of systems and to extend life span. The present study tested the hypothesis that caloric restriction would prevent the previously reported age-related synaptic decline. Accordingly, synaptic density in layer 2 of sensorimotor cortex was compared between 29-month-old male rats fed ad libitum and 29-month-old male rats that were caloric restricted (60% of ad libitum calories) from 4 months of age. In serial electron micrographs, the physical disector was used to determine the numerical density of presumptive excitatory and inhibitory synapses (those containing round or nonround vesicles, respectively) as well as that of neurons. Not only was the previously reported age-related decline in numerical density of presumptive inhibitory synapses not ameliorated by caloric restriction, the numerical density was significantly lower in caloric restricted than in ad libitum fed rats for total as well as for presumptive excitatory and inhibitory synapses. Analysis further revealed no difference in the numerical density of neurons in this region. Relating synapse density to neuron density as the ratio of synapses to neuron also demonstrated significantly fewer synapses per neuron in caloric restricted than in ad libitum fed old rats. Finally, synapse length was significantly less in caloric restricted rats. These results suggest that not only does caloric restriction fail to prevent the age-related decline in presumptive inhibitory synapses, it results in fewer presumptive excitatory synapses as well. PMID- 11897087 TI - NK-1 receptor blockade decreases amphetamine-induced behavior and neuropeptide mRNA expression in the striatum. AB - The effect of intrastriatal administration of LY306740, a specific NK-1 receptor antagonist, on the behavior and changes in gene expression elicited by the psychomotor stimulant, amphetamine, was studied. Acute administration of amphetamine (2.5 mg/kg, i.p.) caused an increase in behavioral activity and preproenkephalin, preprodynorphin and substance P mRNA expression in the striatum. When amphetamine-treated rats were pretreated with LY306740 (35 and 20 nmoles per side, intrastriatally), there was a significant decrease in amphetamine-induced behavioral activity. Quantitative in situ hybridization histochemistry revealed that both concentrations of LY306740 significantly decreased amphetamine-induced mRNA expression of all three neuropeptides. These data indicate that striatal NK-1 receptors modulate amphetamine-induced behavior and mRNA expression of neuropeptides in the rat striatum. PMID- 11897088 TI - Kappa opioid receptor is expressed by somatostatin- and neuropeptide Y-containing interneurons in the rat hippocampus. AB - In our previous studies (J. Chem. Neuroanat. 2000;19:233-241), kappa opioid receptors were immunocytochemically identified in inhibitory interneurons of the dentate hilus and CA1 area of the rat hippocampus. From among the known interneuron subtypes, somatostatin- (SOM) and neuropeptide Y- (NPY) immunoreactive (IR) hippocampal interneurons show morphology and distribution similar to the kappa opioid receptor (KOR) immunopositive cells. In the present study, with the help of double immunocytochemical labelling, we provide direct evidence that the majority of the interneurons immunoreactive for SOM and/or NPY also express the kappa opioid receptor. The receptor was localized on the perikaryal and proximal dendritic region of the SOM- and NPY-immunopositive neurons in the dentate hilus and the CA1 region. From among the SOM immunoreactive cells, 77% in the dentate hilus and 51% in the CA1 stratum oriens was double labelled. In the case of NPY-immunoreactive neurons this proportion was 56 and 65%, respectively. The co-expression of KOR and SOM/NPY suggests that hippocampal interneurons can selectively be activated by the different opioids under different physiological circumstances. PMID- 11897090 TI - Delayed treatment with nicotinamide (vitamin B3) reduces the infarct volume following focal cerebral ischemia in spontaneously hypertensive rats, diabetic and non-diabetic Fischer 344 rats. AB - Since hypertension and/or hyperglycemia are risk factors for stroke, we examined whether the putative neuroprotectant, nicotinamide (NAm), could protect spontaneously hypertensive rats (SHR) or diabetic Fischer 344 rats against focal cerebral ischemia using a model of permanent middle cerebral artery occlusion (MCAo). Intravenous NAm given 2 h after MCAo significantly reduced the infarct volume of SHR (750 mg/kg, 31%, P<0.01) and diabetic (500 mg/kg, 56%, P<0.01) as well as non-diabetic (500 mg/kg, 73%, P<0.01) Fischer 344 rats when compared with saline-injected controls. Thus delayed treatment with NAm protected hypertensive and hyperglycemic rats against a robust model of stroke. PMID- 11897089 TI - Functional expression of TREK-2 K+ channel in cultured rat brain astrocytes. AB - Background K+ channels whose subunit contains four transmembrane segments and two pore-forming domains (4TM/2P) have been cloned recently. We studied whether 4TM/2P K+ channels are functionally expressed in astrocytes that are known to have a large background (resting) K+ conductance and a large resting membrane potential. Reverse transcriptase-PCR analysis showed that, among five 4TM/2P K+ channels examined, TASK-1, TASK-3 and TREK-2 mRNAs were expressed in cultured astrocytes from rat cortex. In cell-attached patches, we mainly observed three K+ channels with single-channel conductances of 30, 117 and 176 pS (-40 mV) in symmetrical 140 mM KCl. The 30 pS channel was the inward rectifying K+ channel that has been previously described in astrocytes. The 117 pS K+ channel also showed inward rectification and was insensitive to 1 mM tetraethylammonium and 1 mM 4-aminopyridine. The 176 pS channel was the Ca2+-activated K+ channel. The 117 pS K+ channel was determined to be TREK-2, as judged by its electrophysiological properties and activation by membrane stretch, free fatty acids and intracellular acidosis. In approximately 50% of astrocytes in culture, whole-cell K+ current increased markedly following application of arachidonic acid. The number of TREK 2 channels in these cells was estimated to be approximately 500-1000/cell. Our results show that TREK-2 is functionally expressed in cortical astrocytes in culture, and suggest that TREK-2 may be involved in K+ homeostasis of astrocytes during pathological states. PMID- 11897091 TI - Regulation of calcitonin gene-related peptide expression in vitro: possibility of a new role for leukemia inhibitory factor. AB - Calcitonin-gene related peptide is among a group of peptides whose expressions are down-regulated following peripheral nerve damage. It is known that this is probably due to deprivation of some target-derived neurotrophic factors, mainly of nerve growth factor though positive effect of other factors, for example that of leukemia inhibitory factor on galanin has also been demonstrated. In this study, the effects of leukemia inhibitory factor and nerve growth factor on calcitonin gene related peptide expression in cultured dorsal root ganglion explants and in their outgrowing axons were examined. Lumbar dorsal root ganglia with short pieces of peripheral nerves were removed from adult mice and explanted into collagen gels. They were covered with RPMI 1640 culture medium and left in an incubator for 2 days after which they were fixed. These whole mount preparations with outgrowing axons were stained with an antibody against calcitonin gene related peptide. Following microscopic examination and imaging, sections were cut from the cultured ganglia as well as from some freshly taken normal ones and they were also stained to determine calcitonin gene related peptide immunoreactivity in the primary sensory neurons. The results demonstrated that besides the positive effect of nerve growth factor on the expression of this peptide in outgrowing axons, leukemia inhibitory factor also supported the expression of calcitonin gene related peptide in the primary sensory neurons of adult mouse lumbar dorsal root ganglia and in their outgrowing axons in vitro. When the time course of changes in calcitonin gene related peptide expression in dorsal root ganglia and the up-regulation of leukemia inhibitory factor at the site of a peripheral nerve injury in vivo are considered together, this novel finding may lead to new explanations for the changes in neuropeptide expression following axotomy. PMID- 11897092 TI - Histological and ultrastructural characterization of interfascicular neurons in the rat anterior commissure. AB - The histological, connectional, and ultrastructural characteristics of a peculiar neuron type in the rat anterior commissure (AC) are described. Since these cells are located among the axonal fascicles of the rostral and caudal parts of the AC, they are termed interfascicular neurons (IFN). In rapid-Golgi sections IFNs appeared in two forms: internuncial (i.e., short axon) and projection neurons (i.e., long axon). The axon of the internuncial neurons terminates upon neighboring IFNs. The projection neurons give rise to an axon which is either incorporated into commissural fibers or ramifies into 12-26 collaterals running laterally in opposite directions along commissural axons. Immunohistochemistry to microtubule-associated protein 2 combined with confocal microscopy showed that IFNs display short varicose dendrites which remain confined to the domain of the AC. The neuronal nature of IFNs was confirmed with the electron microscope on the basis of distinctive organelles and the presence of synaptic inputs. Small areas of neuropil surround some IFNs. These areas are composed of proximal dendrites, terminal axons, axo-shaft and axo-spinous synapses. Because IFNs with their afferents and efferents constitute sufficient elements to integrate neural inputs, it is proposed that they may be involved in processing nerve impulses proceeding from the bilateral cerebral structures connected by the AC. PMID- 11897093 TI - Inhibiting the nucleus tractus solitarii extends cerebrovascular autoregulation during hypertension. AB - Autoregulation maintains cerebral blood flow near basal levels as blood pressure increases, but vasodilation, breakthrough, occurs when hypertension exceeds the autoregulatory range. Loss of breakthrough after transection of baroreceptor nerves suggests that breakthrough is neurally mediated. We hypothesize that central baroreflex interruption will likewise prevent breakthrough. In treated rats, injections of lidocaine into the nucleus tractus solitarii blocked breakthrough and the baroreflex. Therefore, central, like peripheral, baroreflex interruption extends autoregulation during hypertension. PMID- 11897094 TI - Oxidative metabolite of dopamine, 3,4-dihydroxyphenylacetaldehyde, induces dopamine release from PC12 cells by a Ca2+-independent mechanism. AB - 3,4-dihydroxyphenylacetaldehyde (DOPALD), an oxidative metabolite of dopamine (DA), induced dose-dependent DA release from pheochromocytoma (PC12) cells without affecting leakage of lactate dehydrogenase from the cells. DOPALD-induced DA release was independent of extracellular Ca2+ concentration and was not blocked by nifedipine, an L-type Ca2+ channel antagonist. These results indicated a novel intrinsic role of DOPALD in dopaminergic nerve terminals that may take part in the activation of dopamine neurons. PMID- 11897095 TI - Delta opioid receptor localization in the rat cerebellum. AB - Opioid receptors have been localized to a number of brain regions in rats as well as in other species. In situ hybridization has demonstrated the presence of mRNA for the delta receptor subtype in adult rat cerebellar cortex and in several deep nuclei, but there are no reports on localization of the delta receptor protein in cerebellar regions. In the present study, both quantitative immunohistochemistry and Western blots reveal the presence of delta receptors in the adult rat cerebellum, using a specific affinity-purified antibody. Purkinje cells and processes, as well as cells in the granule cell layer, were positively stained with the antibody. Quantitation of confocal microscopy images illustrated a lower relative level of delta receptor immunoreactivity in cerebellar cortical neurons as compared to neurons in hippocampal regions, striatum and cerebral cortex. Stimulation of delta receptors with a selective agonist, DPDPE, in frozen sections of rat brain, induced a significant increase in binding of [35S] GTPgammaS in the cerebellar cortex as compared to basal binding levels, thereby demonstrating coupling of the receptor subtype to G-protein. Functional implications for the delta receptor in the cerebellum are discussed, particularly in light of evidence for the presence of a cerebellar opioid receptor for the endogenous opioid methionine enkephalin during early postnatal life. PMID- 11897096 TI - Pathways involved in synchronization of sympathetic nerve discharge to lung inflation. AB - In urethane-anesthetized cats, we tested the hypothesis that sympathetic nerve discharge (SND) is synchronized to lung inflation, in part, over a vagal afferent pathway that bypasses the central respiratory oscillator. For this purpose, partial coherence analysis was used to mathematically remove the portion of the relationship between intratracheal pressure (ITP; index of lung inflation) and inferior cardiac SND that was common to that between ITP and phrenic nerve activity (PNA; index of central respiratory cycle). ITP-SND partial coherence (PNA-related component removed) at the frequency of breathing should be significantly different from zero if a bypass pathway exists. This was found to be the case in paralyzed, artificially ventilated cats when lung inflation and rhythmic PNA were coupled in various ratios (e.g. 1:1, 3:1) and in spontaneously breathing cats. As expected, surgical vagotomy eliminated ITP-SND coherence in artificially ventilated cats. These findings support the view that a portion of the relationship between ITP and SND is independent of the influences of pulmonary stretch receptors on central respiratory networks. PMID- 11897097 TI - Induction of cytochrome c-mediated apoptosis by amyloid beta 25-35 requires functional mitochondria. AB - Accumulating data suggest a central role for mitochondria and oxidative stress in neurodegenerative apoptosis. We previously demonstrated that amyloid-beta peptide 25-35 (Abeta 25-35) toxicity in cultured cells is mediated by its effects on functioning mitochondria. In this study, we further explored the hypothesis that Abeta 25-35 might induce apoptotic cell death by altering mitochondrial physiology. Mitochondria in Ntera2 (NT2 rho+) human teratocarcinoma cells exposed to either staurosporine (STS) or Abeta 25-35 were found to release cytochrome c, with subsequent activation of caspases 9 and 3. However, NT2 cells depleted of mitochondrial DNA (rho0 cells), which maintain a normal mitochondrial membrane potential (Deltapsi(m)) despite the absence of a functional electron transport chain (ETC), demonstrated cytochrome c release and caspase activation only with STS. We further observed increased reactive oxygen species (ROS) production and decreased reduced glutathione (GSH) levels in rho+ and rho0 cells treated with STS, but only in rho+ cells treated with Abeta 25-35. We conclude that under in vitro conditions, Abeta can induce oxidative stress and apoptosis only when a functional mitochondrial ETC is present. PMID- 11897098 TI - Temporal changes in the neurotrophic environment of the denervated striatum as determined by the survival and outgrowth of grafted fetal dopamine neurons. AB - There is growing evidence that the neurotrophic environment of the denervated striatum may change with time following a lesion of the nigrostriatal pathway in young adult rats. To test this hypothesis, we implanted fetal dopamine grafts into the striatum at several different time points relative to the nigrostriatal pathway lesion and allowed the grafts to integrate with the host for a period of 1 month; subsequently, we observed the function and morphology of the dopamine grafts. Fetal grafts were implanted at the following time points relative to the lesion: 1 week before (-1 Week), at the same time (Week 0), 1 week after (1 Week), 4 weeks after (4 Weeks), or 12 weeks after (12 Weeks). Amphetamine-induced rotational behavior was assessed 4 weeks after grafting for all groups. Rotational scores indicate that grafts for the 1 Week group showed the greatest reversal of amphetamine-induced rotational behavior that was also significantly greater than the scores for the -1 Week group. Morphological analysis revealed that grafts in the Week 0, 1 Week and 4 Weeks groups showed a significantly larger area of tyrosine hydroxylase-positive (TH+) fiber outgrowth than in the -1 Week group, while fiber outgrowth for the 12 Weeks group was significantly lower than for the 1 Week group. Cell count analysis for TH+ neurons within the graft indicate a significantly greater number of TH+ neurons in grafts for the 1 Week group than in grafts for the -1 Week. The results of this study suggest that neurotoxic lesions may induce a compensatory increase in neurotrophic activity within the denervated striatum of young rats that is conducive to the survival and outgrowth of fetal dopamine grafts. These data also correlate well with reports that the expression of several specific dopaminergic neurotrophic factors within the striatum increase following a neurotoxic lesion of the nigrostriatal pathway in young adult rats. PMID- 11897099 TI - Excitotoxic neurodegeneration induced by intranasal administration of kainic acid in C57BL/6 mice. AB - Glutamate excitotoxicity plays a key role in inducing neuronal cell death in many neurological diseases. In mice, administration of kainic acid, an analogue of the excitotoxin glutamate, results in hippocampal cell death and seizures. Kainic acid-induced seizures in mice provide a well-characterized model for studies of human neurodegenerative diseases. However, C57BL/6 mice, which are often used for genetic analyses and transgenic and knockout studies, are resistant to excitotoxicity induced by subcutaneous administration of kainic acid. In the present study, kainic acid administered by the intranasal route was shown to result in continuous tonic-clonic seizures in C57BL/6 mice. These seizures continued for 1-5 h and successfully induced selective lesions in area CA3 of the hippocampus. The survival rate was high even after mice experienced severe seizures. The hippocampal lesions were associated with a high level of cyclooxygenase-2 production as well as astrogliosis. Administration of kainic acid also altered behavioral responses, with mice showing a significant increase in locomotion and rearing activity as indicated by an open-field test. This animal model could provide a valuable tool for exploring the role of excitotoxicity in neuropathological conditions and should be further evaluated in gene-targeting studies of neurodegenerative diseases. PMID- 11897100 TI - Effect of hyperthermia on the transport of mRNA encoding the extracellular matrix glycoprotein SC1 into Bergmann glial cell processes. AB - SC1 is an extracellular matrix glycoprotein that is related to the multifunctional protein SPARC. These matricellular members play regulatory roles in modulating cellular interactions. SC1 expression is enriched in the central nervous system during embryonic and postnatal development as well as in the adult brain. In the rat cerebellum, SC1 is expressed at high levels in Bergmann glial cells and their radial fibers which project into the synaptic-rich molecular layer. At specific stages of development and in the adult, SC1 mRNA is selectively transported into cellular processes of these cells. In the present study, we have examined the effect of whole-body hyperthermia on the transport of SC1 mRNA in Bergmann glial cells of the rat cerebellum. Our results show that SC1 mRNA transport is diminished at 10 and 15 h post-hyperthermia, but returns to control levels by 24 h after heat shock. One of the characteristics of a heat shock on cells grown in tissue culture is a collapse of the cytoskeletal network. Intact components of the cytoskeleton are necessary for the transport of mRNA into peripheral processes of cells. However, in vivo hyperthermia does not appear to affect the morphology of the intermediate filament proteins GFAP, vimentin, or the beta-tubulin component of microtubules in Bergmann glial cell processes. During the hyperthermic time course, levels of vimentin protein increase, which is reflected by immunoreactivity of activated astrocytes and microvasculature in cerebellar white matter. PMID- 11897101 TI - Fos-like immunoreactivity and thirst following hyperosmotic loading in rats with subdiaphragmatic vagotomy. AB - If receptors in the gut relay information about increases in local osmolality to the brain via the vagus nerve, then vagotomy should diminish this signaling and reduce both thirst and brain Fos-like immunoreactivity (Fos-ir). Water intake in response to hypertonic saline (i.p. or i.g., 1 M NaCl, 1% BW; i.g., 0.6 M NaCl, 0.5% BW) was reduced during 120 min in rats with subdiaphragmatic vagotomy (VGX) compared to sham-VGX rats. Brain Fos-ir was examined in response to both i.g. loads. After the smaller load, VGX greatly reduced Fos-ir in the supraoptic nucleus (SON) and the magnocellular and parvocellular areas of the paraventricular nucleus (PVN). Fos-ir in the subfornical organ (SFO) and nucleus of the solitary tract (NTS) was not affected. After the larger load, VGX significantly reduced Fos-ir in the parvocellular PVN and in the NTS, but not in the other regions. Thus, decreased water intake by VGX rats was accompanied by decreased Fos-ir in the parvocellular PVN after the same treatments, indicating a role for the abdominal vagus in thirst in response to signaling from gut osmoreceptors. The decreased water intake in the VGX group was not reflected as a decrease in Fos-ir in the SFO. Absorption of the larger i.g. load may have activated Fos-ir through more rapidly increasing systemic osmolality, thereby obscuring a role for the vagus at this dose in the SON and magnocellular PVN. PMID- 11897102 TI - Bilirubin produces apoptosis in cultured bovine brain endothelial cells. AB - Blood components such as oxyhemoglobin are believed to cause cerebral vasospasm by inducing contraction and cell death in cerebral arteries. We have observed previously that oxyhemoglobin produces apoptotic changes in cultured endothelial cells. This study was undertaken to explore if bilirubin, a bi-product of hemoglobin degradation, will produce similar cytotoxicity in endothelial cells. Cultured bovine brain microvascular endothelial cells were incubated in four concentrations of bilirubin (10, 25, 50, and 100 microM) for varying times (6, 12, and 24 h). Control cells were incubated in saline or vehicle (NaOH solution, <0.01% of 0.01 N) for similar time periods. The cultured cells were then observed microscopically for evidence of cellular alterations. Bilirubin (10-100 microM) produced apoptosis that appeared time-dependent but not clearly concentration dependent. Biochemical markers for apoptosis such as DNA fragmentation and PARP cleavage were induced by bilirubin. We conclude that endothelial cells may undergo apoptosis after exposure to bilirubin. PMID- 11897104 TI - Antioxidant compounds protect dopamine neurons from death due to oxidative stress in vitro. AB - Using tissue culture models of oxidative stress caused by serum deprivation or MPTP/MPP+ toxicity, the present study establishes that the antioxidants epigallocatechin gallate, lazaroids U74389G and U83836E, reservatrol, MnTBAP, MCI 186, trolox, and melatonin protect 68-100% of dopamine (DA) neurons from cell death. In contrast, the nitric oxide inhibitor LY83583, the caspase inhibitors Z VAD-FMK, Ac-DQMD-CHO and Z-DEVD-FMK, and the CDK-5 inhibitor, roscovotine were not neuroprotective, although death was often delayed by 1 day in vitro. We conclude that antioxidants are more effective at preventing cell death in vitro than are inhibitors at later stages in the death cascade. PMID- 11897103 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin treatment induces c-Fos expression in the forebrain of the Long-Evans rat. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is one of the most toxic environmental pollutants. In the present study, we examined c-Fos expression in the central nervous system (CNS) after administration of a lethal dose of TCDD to the adult Long-Evans rat to clarify if the CNS participates in TCDD-induced intoxication. A single dose of TCDD (dissolved in olive oil, 50 microg/kg) or olive oil alone was administered to the rats by gavage. Animals were allowed to survive for 1 day to 5 weeks. Three days after the administration, a significantly large number of Fos immunopositive cells were found in the hypothalamus (i.e. dorsomedial hypothalamic nucleus, paraventricular hypothalamic nucleus, medial preoptic nucleus), central amygdaloid nucleus and bed nucleus of the stria terminalis. These results suggest that some TCDD toxicity may be induced by its direct action on the CNS. PMID- 11897105 TI - Na+, K+ ATPase alpha-subunit isoform distribution and abundance in guinea-pig longitudinal muscle/myenteric plexus after exposure to morphine. AB - Previous work in the myenteric plexus has shown that the resting membrane potential of morphine-tolerant guinea-pig myenteric S neurons is significantly depolarized relative to placebo-implanted controls, and that this depolarization is associated with reduced electrogenic Na+, K+ pumping. Identification of the subunits of the sodium pump which are in the myenteric plexus was undertaken in order to facilitate direct qualitative and quantitative measurements of the abundance of sodium pump isoforms after morphine exposure, thereby confirming and extending the electrophysiological data to the molecular level. Seven days prior to the experiments, tolerance was induced by subcutaneous implantation of morphine pellets (one pellet, 75 mg/100 g body weight) while control guinea pigs received placebo pellets. Using immunohistochemistry and confocal microscopy, the distribution of the alpha subunit isoforms of the Na+/K+ -ATPase in placebo and morphine-tolerant guinea-pig ileum was determined. Only the alpha1 and alpha3 subunit isoforms were in sufficient abundance to be observed. The alpha1 subunit isoform was most highly concentrated in the mucosa and in neurons. In contrast, the alpha3 subunit isoform was uniquely localized to neurons. Western and slot blot analyses of longitudinal muscle/myenteric plexus homogenates identified a significant reduction of the alpha3 but not the alpha1 subunit isoform in tolerant preparations. It is concluded that the reduced electrogenic pumping in the S neurons after morphine exposure is associated with a reduction in the alpha3 subunit isoform. PMID- 11897106 TI - Age-related changes in the distribution of nitrotyrosine in the cerebral cortex and hippocampus of rats. AB - A wealth of indirect evidence implicates oxidative damage of cellular constituents in aging, as well as in the pathogenesis of the neurodegenerative diseases of later years. In the present study, we have determined age-related changes in the distribution of 3-nitrotyrosine (3-NT) in the cerebral cortex and hippocampus of rats. In adult rats, no 3-NT-immunoreactive cells were found in the cerebral cortex and hippocampus, whereas 3-NT immunoreactivity was significantly increased in aged rats. Some pyramidal cells of CA3 area and granule cells of the dentate gyrus highly expressed 3-NT in aged rats. Many interneurons located within stratum pyramidale and stratum oriens of CA1 were strongly immunoreactive for 3-NT. Our first demonstrations of increased 3-NT in the cerebral cortex and hippocampus during aging implicate these areas as sites for functionally significant oxidative damage. The mechanisms underlying the increased immunoreactivity for 3-NT, and the functional implications of this increase, require elucidation. PMID- 11897107 TI - Group I metabotropic glutamate receptors activate burst firing in rat midbrain dopaminergic neurons. AB - We have investigated the changes in the spontaneous firing pattern induced by DHPG ((S)-3,5-dihydroxyphenylglycine) and NMDA (N-methyl-d-aspartic acid) on rat dopaminergic neurons in substantia nigra pars compacta (SNc) using sharp microelectrode recordings in in vitro conditions. Twenty-five out of 33 cells modified the regular single-pacemaker activity in burst firing when exposed to the Group I metabotropic glutamate receptor (mGluR) agonist DHPG (30 microM) and d-tubocurarine (500 microM) (d-TC), whereas they all fired in bursts during NMDA (20 microM) plus d-TC application. The blockade of SK-channels by d-TC and apamin was essential for the production of both types of bursts. Although the two drugs induced a similar number of action potentials per burst, the DHPG-induced bursts had a lower frequency, a longer duration and a longer plateau period without spikes. In addition, the DHPG-induced bursting had a longer wash-out, could be reduced or blocked by the mGluR 1 selective, non-competitive antagonist CPCCOEt (7-cyclopropan[b]chromen-1a-carboxylic acid ethyl ester) (100 microM) while it was not affected by the mGluR 5 selective antagonist MPEP (2-methyl-6 (phenylethynyl)-pyridine (10 microM). These results suggest that both the activation of glutamate metabotropic type 1 and NMDA ionotropic receptors induce burst firing in the dopaminergic cells of the ventral midbrain when the activity of the SK-channels is reduced. PMID- 11897108 TI - Presynaptic group II metabotropic glutamate receptors reduce stimulated and spontaneous transmitter release in human dentate gyrus. AB - Metabotropic glutamate receptors (mGluRs) control excitatory neurotransmission as inhibitory autoreceptors at many synapses throughout the CNS. Since pharmacological activation of mGluRs potently depresses excitatory transmission, anticonvulsive effects were found in a number of experimental epilepsies. However, although native rodent mGluRs and heterologously expressed human mGluRs have so far been investigated in great detail, our knowledge about native human mGluRs in situ is limited. Here we used acute human hippocampal slices prepared from hippocampi surgically removed for the treatment of temporal lobe epilepsy in order to investigate the modulation of glutamatergic transmission by human mGluRs at the perforant path-granule cell synapse. The broad spectrum mGluR agonist (1S, 3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) profoundly and reversibly reduced field EPSPs (fEPSPs) with an EC(50) of 30+/-7.4 microM. Paired-pulse depression of fEPSPs was converted into strong facilitation. The inhibition of fEPSPs by ACPD was mimicked by the specific group II mGluR agonist (2S, 2'R, 3'R) 2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV), while the specific group I agonist (S)-3,5-dihydroxyphenylglycine (DHPG) was ineffective. The effect of ACPD was blocked by group II antagonist (2S,3S,4S)-2methyl-2 (carboxycyclopropyl)glycine (MCCG) but was not changed by coapplication of the specific group III antagonist (S)2 amino2methyl4phosphonobutanoic acid (MAP4). ACPD reduced pharmacologically isolated intracellular EPSPs in granule cells to the same extent as fEPSPs, whereas a specific group III agonist had no effect on EPSPs. Whole-cell recordings from morphologically identified granule cells revealed that DCG-IV significantly reduced the frequency of miniature EPSCs (mEPSCs) in granule cells while the mean amplitude of mEPSCs was not affected. We conclude that in human dentate gyrus mGluR2/3 can almost completely depress glutamate release by a presynaptic mechanism which acts downstream of presynaptic voltage gated calcium-entry and most likely involves a direct modulation of the release machinery. PMID- 11897109 TI - Functional expression of distinct NMDA channel subunits tagged with green fluorescent protein in hippocampal neurons in culture. AB - We generated expression vectors for N-terminally green fluorescent protein tagged NR2A and NR2B subunits (GFP-NR2A and GFP-NR2B). Both constructs expressed GFP and formed functional NMDA channels with similar properties to untagged controls when co-transfected with NR1 subunit partner in HEK293 cells. Primary cultured hippocampal neurons were transfected at five days in vitro with these vectors. Fifteen days after transfection, well-defined GFP clusters were observed for both GFP-NR2A and GFP-NR2B subunits being co-localized with endogenous NR1 subunit. Whole-cell recordings showed that the GFP-NR2A subunit determined the decay of NMDA-mediated miniature spontaneous excitatory postsynaptic currents (NMDA-mEPSCs) in transfected neurons. Live staining with anti-GFP antibody demonstrated the surface expression of GFP-NR2A and GFP-NR2B subunits that was partly co-localized a presynaptic marker. Localization of NMDA receptor clusters in dendrites was studied by co-transfection of CFP-actin and GFP-NR2 subunits followed by anti-GFP surface staining. Within one week after plating most surface NMDAR clusters were distributed on dendritic shafts. Later in development, a large portion of surface clusters for both GFP-NR2A and GFP-NR2B subunits were clearly localized at dendritic spines. Our report provides the basis for studies of NMDA receptor location together with dendritic dynamics in living neurons during synaptogenesis in vitro. PMID- 11897110 TI - Studies on the subtype selectivity of CP-101,606: evidence for two classes of NR2B-selective NMDA receptor antagonists. AB - The subtype-selectivity of racemic [(3)H]CP-101,606, a novel high-affinity NMDA receptor radioligand was determined using defined recombinant NMDA receptor subunits expressed in HEK 293 cells. [(3)H]CP-101,606 binds to adult rodent forebrain and NR1/NR2B receptors expressed in HEK 293 cells with K(D)=4.2 nM and 6.0 nM, respectively. In contrast, no high affinity specific binding was detected to NR1, NR2A, NR2B subunits expressed alone or NR1/NR2A receptors. HEK 293 cells were transfected with NR1, NR2A and NR2B receptor subunits and complexes comprising all three subunits were isolated by anti-NR2A immunoaffinity chromatography. Based on immunoblotting with subunit-selective antibodies, the immunopurified material contained all three NMDA receptor subunit polypeptides. However, in contrast to parallel studies in which high affinity [(3)H]Ro-25,6981 binding activity was observed, no high affinity [(3)H]CP-101,606 binding sites were detected to the immunopurified material. This study provides further evidence for two distinct classes of NR2B-directed NMDA receptor antagonists, one which binds with high affinity irrespective whether another NR2 subunit type is present (Ro-25,6981) and a second class which is affected significantly by the presence of another NR2 subunit type within the receptor complex, exemplified by CP-101,606. PMID- 11897111 TI - Changes in GABAA receptor gamma 2 subunit gene expression induced by long-term administration of oral contraceptives in rats. AB - The effects of oral contraceptives (OCs) on neurosteroid concentrations were evaluated in female rats and women. In rats, ethynylestradiol and levonorgestrel (0.030 and 0.125 mg, respectively, subcutaneously) administered daily for 6 weeks reduced the concentrations of pregnenolone (-41%) progesterone (-74%) and allopregnanolone (-79%) in the cerebral cortex; the plasma concentrations of these steroids were also reduced but by smaller extents. OC administration for 3 months also reduced the serum concentrations of pregnenolone, progesterone and allopregnanolone in women. Chronic administration of OCs in rats increased the abundance of gamma-aminobutyric acid type A (GABA(A)) receptor gamma 2L and gamma 2S subunit mRNAs and the relative protein in the cerebral cortex, while the amounts of various alpha and beta subunit mRNAs were unaffected. Ovariectomy did not modify the effect of OCs administration on the concentrations of neurosteroids in the rat cerebral cortex (but not in the plasma) as well as on the GABA(A) receptor gene expression, suggesting a direct effect of OCs in brain. Finally, rats treated with OCs exhibited an anxiety-like behavior in the elevated plus-maze test. These results indicate that long-term treatment with OCs induced a persistent reduction in the concentrations of pregnenolone, progesterone and its GABA(A) receptor-active metabolite, allopregnanolone, both in rats and women. In rats this effect was associated with a plastic adaptation of GABA(A) receptor gene expression in the rat cerebral cortex. PMID- 11897112 TI - Volatile anesthetic actions on the GABAA receptors: contrasting effects of alpha 1(S270) and beta 2(N265) point mutations. AB - Previous studies have suggested that two specific amino acid residues in transmembrane segments 2 and 3 of the GABA(A) receptor alpha 2 subunit, Ser270 and Ala291, are critical for the enhancement of GABA(A) receptor function by inhaled anesthetics. The aim of this study was to determine the effects of amino acid substitutions in alpha 1 beta 2 gamma 2s GABA(A) receptors at alpha 1(S270) and at the homologous beta 2(N265) on receptor gating and anesthetic potentiation of GABA-induced responses. The wild-type and mutant receptors were transiently expressed in HEK 293 cells and GABA-induced currents were recorded using whole cell voltage clamp. Potentiation of responses to a submaximal concentration of GABA by the anesthetics halothane and isoflurane was also examined. Some of the point mutations caused shifts in the GABA dose-response curve, indicating that the mutations changed the apparent affinity of the receptor for GABA. In receptors mutated at alpha 1(S270), the GABA EC(50) is inversely correlated with the volume of the residue of 270. On the contrary, there was no clear relationship between the physical properties of the amino acid residue at 265 in the beta 2 subunit and either the GABA EC(50) or anesthetic modulation, although mutations at N265 altered both parameters in a quantitative manner. These data are consistent with the results of previous work using other subunit combinations, in confirming that alpha 1(S270) may be involved in channel gating, and also may be important in anesthetic binding; the role of beta 2(N265) is less clear. PMID- 11897113 TI - Levetiracetam opposes the action of GABAA antagonists in hypothalamic neurones. AB - Whole-cell current-clamp recordings in guinea-pig brain slices were used to assess the effect of the novel antiepileptic drug levetiracetam (LEV; Keppra) on the gamma-aminobutyric acid-A (GABA(A)) responses evoked by exogenous applications of the agonists GABA and muscimol on hypothalamic neurones. LEV (40 microM) had no direct effect on GABA(A) responses but it occluded the GABA(A) receptor blocking action of bicuculline-methiodide (100 microM) and, to a lesser extent, the GABA(A)-receptor blocking action of gabazine (50 microM). While previous reports have indicated that the inhibition by LEV of the epileptiform hyperexcitability induced by bicuculline in rat hippocampus might occur via non GABAergic mechanisms, the present data suggest a possible indirect modulation by LEV of GABA-gated currents in guinea-pig hypothalamic neurones. PMID- 11897114 TI - Gabapentin-mediated inhibition of voltage-activated Ca2+ channel currents in cultured sensory neurones is dependent on culture conditions and channel subunit expression. AB - We have used the whole cell patch clamp method and fura-2 fluorescence imaging to study the actions of gabapentin (1-(aminoethyl) cyclohexane acetic acid) on voltage-activated Ca(2+) entry into neonatal cultured dorsal root ganglion (DRG) neurones and differentiated F-11 (embryonic rat DRG x neuroblastoma hybrid) cells. Gabapentin (2.5 microM) in contrast to GABA (10 microM) did not influence resting membrane potential or input resistance. In current clamp mode gabapentin failed to influence the properties of evoked single action potentials but did reduce the duration of action potentials prolonged by Ba(2+). Gabapentin attenuated high voltage-activated Ca(2+) channel currents in a dose- and voltage- dependent manner in DRG neurones and reduced Ca(2+) influx evoked by K(+) depolarisation in differentiated F-11 cells loaded with fura-2. The sensitivity of DRG neurones to gabapentin was not changed by the GABA(B) receptor antagonist saclofen but pertussis toxin pre-treatment reduced the inhibitory effects of gabapentin. Experiments following pre-treatment of DRG neurones with a PKA activator and a PKA-inhibitor implicated change in phosphorylation state as a mechanism, which influenced gabapentin action. Sp- and Rp-analogues of cAMP significantly increased or decreased gabapentin-mediated inhibition of voltage activated Ca(2+) channel currents. Culture conditions used to maintain DRG neurones and passage number of differentiated F-11 cells also influenced the sensitivity of Ca(2+) channels to gabapentin. We analysed the Ca(2+) channel subunits expressed in populations of DRG neurones and F-11 cells that responded to gabapentin had low sensitivity to gabapentin or were insensitive to gabapentin, by Quantitative TaqMan PCR. The data obtained from this analysis suggested that the relative abundance of the Ca(2+) channel beta(2) and alpha(2)delta subunit expressed was a key determinant of gabapentin sensitivity of both cultured DRG neurones and differentiated F-11 cells. In conclusion, gabapentin inhibited part of the high voltage-activated Ca(2+) current in neonatal rat cultured DRG neurones via a mechanism that was independent of GABA receptor activation, but was sensitive to pertussis toxin. Gabapentin responses identified in this study implicated Ca(2+) channel beta(2) subunit type as critically important to drug sensitivity and interactions with alpha(1) and alpha(2)delta subunits may be implicated in antihyperalgesic therapeutic action for this compound. PMID- 11897116 TI - The role of 5-HT1B and 5-HT1D receptors in the selective inhibitory effect of naratriptan on trigeminovascular neurons. AB - The importance of 5-HT(1B) and 5-HT(1D) receptors in the actions of the anti migraine drug naratriptan was investigated using the relatively selective 5-HT(1) receptor ligands SB224289 and BRL15572. Electrical stimulation of the superior sagittal sinus (SSS) in cats activated neurones in the trigeminal nucleus caudalis. Facial receptive fields (RF) were also electrically stimulated to activate the same neurones. Responses of these neurones to SSS stimulation were suppressed by iontophoretic application of naratriptan (5-50 nA). There were two distinct populations of neurones in the nucleus--those in deeper laminae in which the responses to SSS and RF stimulation were equally suppressed by naratriptan ('non-selective') and more superficial neurones in which only the SSS responses were suppressed by naratriptan ('selective'). Concurrent micro-iontophoretic application (50 nA) of the 5-HT(1D) antagonist BRL15572 antagonised the suppression by naratriptan of the response of 'selective' cells to SSS stimulation. Iontophoretic application of SB224289 (50 nA), a 5-HT(1B) antagonist, antagonised the suppression by naratriptan of responses of 'non selective' cells to RF stimulation and, to a lesser extent, also antagonised the suppression of responses to SSS stimulation. Intravenous administration of SB224289 antagonised the suppression only of RF responses of "non-selective" neurons by naratriptan and intravenous administration of BRL15572 antagonised the suppression only of SSS responses of "selective" neurons by naratriptan. These results suggest that the response of nucleus caudalis neurons to stimulation of the sagittal sinus can be modulated by both 5-HT(1B) and 5-HT(1D) receptor activation, with the 5-HT(1D) receptors perhaps playing a greater role. The response to RF stimulation is more influenced by 5-HT(1B) receptor modulation with 5-HT(1D) receptors being less important. Therefore, this suggests that selective 5-HT(1D) agonists may be able to target the neuronal population, which is selectively involved in the transmission of dural inputs. We conclude that the central terminals of trigeminal primary afferent fibres contain 5-HT(1B) and 5 HT(1D) receptors. Primary afferents from the dura mater may predominantly express 5-HT(1D) receptors, while facial afferents may predominantly express 5-HT(1B) receptors. Activation of 5-HT(1D) receptors in particular may be important in the anti-migraine effect of naratriptan. PMID- 11897117 TI - ATP-gated currents in rat primary auditory neurones in situ arise from a heteromultimetric P2X receptor subunit assembly. AB - Spiral ganglion neurones provide the primary afferent innervation to sensory hair cells within the mammalian cochlea. Recent evidence suggests that their function may be modulated by purinergic signalling mechanisms, associated with release of adenosine 5'-triphosphate (ATP). Utilising a newly developed slice preparation of the neonatal rat cochlea, we have investigated the response of neurones in situ, to purinergic agonists and antagonists using whole-cell voltage clamp recordings. In cells identified as type I spiral ganglion neurones on the basis of morphology and voltage-dependent conductances, pressure-applied ATP, alpha,beta-methyleneATP (alpha,beta-meATP), 2-methylthioATP (2-MeSATP) and adenosine 5'-diphosphate (ADP) elicited a consistent phenotype of desensitising, inwardly rectifying current. The ATP-activated currents were reversibly blocked by the P2X receptor antagonists pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS, 10 microM), and 2',3'-O-(2,4,6-trinitrophenyl)-ATP (TNP-ATP; IC(50) 407 nM). Neurones were more sensitive to ATP at low pH. The EC(50) value for ATP shifted from 18 microM at pH 7.3, to 1 microM at pH 6.3, with Hill coefficients of approximately 1. The results indicate that ATP-gated ion channels in spiral ganglion neurones arise from a specific heteromultimeric assembly of P2X receptor subunits which has no correspondence with present recombinant P2X receptor models. PMID- 11897115 TI - Continuous subcutaneous infusion of apomorphine rescues nigro-striatal dopaminergic terminals following MPTP injection in mice. AB - Apomorphine has been introduced in the treatment of late-stage Parkinson's Disease (PD). The disadvantage of a short half-life of apomorphine is now overcome by the use of a continuous subcutaneous (s.c.) self-delivering system. We examined whether continuous s.c. infusion of apomorphine rescues nigro striatal dopaminergic neurons from toxicity induced by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) in mice. Apomorphine was continuously infused in mice by means of a s.c. minipump that delivered the drug at a rate of 0.5 or 3.15 mg/kg/day. MPTP induced a >80% reduction in striatal dopamine (DA) after one day. DA levels were still substantially reduced one month following MPTP injection, in spite of a partial recovery. Similarly, striatal immunoreactivity for tyrosine hydroxylase and dopamine transporter was markedly reduced at this time interval. Continuous s.c. infusion of apomorphine starting 40 h following MPTP injection rescued striatal dopaminergic terminals, as assessed by measurements of DA and its metabolites, as well as TH and DAT immunostaining after one month. The neurorescuing effect was more remarkable at a delivery rate of 3.15 mg/kg/day of apomorphine. In contrast, no rescue was observed when apomorphine was administered as a single daily s.c. bolus of 1 or 5mg/kg starting 40 h following MPTP. We conclude that apomorphine is able to rescue nigro-striatal dopaminergic neurons when continuously delivered at doses that are comparable to those delivered by minipumps in PD patients. These results suggest that continuous s.c. infusion of apomorphine not only relieves the symptoms, but also reduce the ongoing degeneration of nigro-striatal dopaminergic neurons in PD patients. PMID- 11897118 TI - Somatostatin sst2 receptor knock-out mice: localisation of sst1-5 receptor mRNA and binding in mouse brain by semi-quantitative RT-PCR, in situ hybridisation histochemistry and receptor autoradiography. AB - The peptide hormone/neurotransmitter somatostatin (somatotropin release inhibiting factor; SRIF) and its receptors (sst(1)-sst(5)) appear to regulate many physiological functions in the CNS. Semi-quantitative analysis of the densities of mRNA expression for sst(1-5) receptors and SRIF receptor binding sites were established in sst(2) receptor knock-out (KO) mice. Patterns of sst(1 5) receptor mRNA expression were largely conserved for sst(1,3,4) and sst(5) selective oligonucleotide probes; whereas sst(2) signals were completely absent in KO mouse brain. Autoradiographic analysis demonstrated [(125)I]LTT SRIF(28), [(125)I]CGP 23996 (two radioligands known to label all five recombinant SRIF receptors) and [(125)I]Tyr(3)-octreotide (sst(2) and sst(5) receptor selective) binding in wild type (WT) mouse brain sections; yet no specific binding of [(125)I]Tyr(3)-octreotide in KO mice. In contrast, [(125)I]LTT SRIF(28) and [(125)I]CGP 23996 binding was still present in a number of brain areas in KO mice, although to a lesser degree than in those regions where [(125)I]Tyr(3) octreotide binding was found, in WT animals. The present data suggest first, that both sst(2) receptor protein and mRNA were completely absent in the brain of these KO animals. Second, there was little evidence of compensatory regulation, at the mRNA level, of the other SRIF receptors as a consequence of the sst(2) KO. Third, the absence of any [(125)I]Tyr(3)-octreotide binding, in KO mice, suggests that this particular ligand is selective for the sst(2) receptor subtype (under the conditions utilised); or that sst(5) receptors are only marginally expressed in brain. Fourth, there were regions where the binding of [(125)I]LTT SRIF(28) and [(125)I]CGP 23996 were moderately affected by the sst(2) KO, suggesting that additional SRIF receptors may well contribute to the binding of the aforementioned radioligands. Finally, since the relative distribution of these two ligands were not entirely superimposable, it suggests that their respective selectivity profiles towards the different SRIF receptor subtypes in situ are not identical. PMID- 11897119 TI - Neonatal nitric oxide synthase inhibition: social interaction deficits in adulthood and reversal by antipsychotic drugs. AB - Nitric oxide synthase (NOS) is thought to migrate improperly during development in the brains of schizophrenic patients. Also it is known that nitric oxide (NO) effects synaptogenesis during development of the CNS. Previously we have shown that neonatal treatment with a NOS inhibitor effects an animal's sensitivity to amphetamine and PCP. In the present study, neonatal rats were challenged with a NOS inhibitor (L-nitroarginine, 10mg/kg, s.c.) daily on post-natal days (PD) three, four and five. L-Nitroarginine (L-NoArg) treated male rats at adulthood (PD56 and older) had a deficit in social interaction (SI) when placed in an environment with another foreign male rat and this deficit was reproducible on a weekly basis for at least five weeks. Haloperidol failed to significantly reverse this deficit before pronounced secondary effects on general behavior were seen at high doses. However, the atypical antipsychotics, clozapine and olanzapine, were able to significantly reverse this deficit at doses which did not effect baseline SI values. In a separate cohort of animals the effect of DOI was investigated, this was done to ascertain if there was a differential sensitivity of serotonergic pathways in this model. There was no difference in the behavioral score elicited from control or NoArg-treated rats. It is suggested that the SI deficits seen here may be more sensitive to atypical antipsychotics rather than haloperidol. PMID- 11897120 TI - Neonatal maternal deprivation modifies feeding in response to pharmacological and behavioural factors in adult rats. AB - Neonatal maternal deprivation permanently modifies the hypothalamo-pituitary adrenal (HPA) axis and other neurobiological and behavioural parameters in rats. The HPA axis plays a central role in the control of feeding, and participates in the anorexigenic action of dexfenfluramine and restraint stress, and in the orexigenic action of a cafeteria diet. Therefore, we investigated whether maternal deprivation modifies feeding responses to these factors. Experimental pups were separated for 24h from the mother 5 or 14 days after birth. The anorexigenic response to both dexfenfluramine and restraint stress was increased, and body weight as well as subcutaneous adipose tissue gain induced by cafeteria diet was higher in early deprived adult rats. However, these effects were dependent on the time of maternal deprivation. According to our predictions, the feeding response of maternally deprived rats to anorexigenic and orexigenic agents was altered, which is probably partly due to an altered HPA function, but the participation of the serotonergic, the opioid and/or the dopaminergic system cannot be ruled out. Additional studies are needed to detail precisely the neurobiological substrates of modified feeding behaviour of maternally deprived animals. This early stress paradigm altering feeding behaviour could become an interesting model for research into human eating disorders. PMID- 11897121 TI - The COX inhibitors indomethacin and meloxicam exhibit anti-emetic activity against cisplatin-induced emesis in piglets. AB - We analysed the effects of four cyclooxygenases (COX) inhibitors on cisplatin induced emesis in piglets. Ninety-five animals receiving cisplatin (5.5 mg kg( 1), i.v.) were observed for 60 h. One hour prior to cisplatin, controls (n=29) were dosed with a saline solution while experimental animals received an i.v. or i.p. injection of one of the COX inhibitors. Additional injections of COX inhibitor were given at 15 and 39 h after cisplatin administration (or every 6h in one group receiving diclofenac). The latencies to the first emetic episode (EE) compared to controls (2.1+/-0.4 h) increased in groups receiving naproxen (4.66+/-0.94 h, n=9, 30 mg kg(-1)) and indomethacin (6.19+/-1.13 h, n=7, 10 mg kg(-1)) i.v. Indomethacin significantly decreased the incidence of both the acute (by 40%) and delayed (by 66%) phases of emesis. The total number of EE during the 60 h compared to controls (28.3+/-1.9 EE) was significantly reduced in piglets receiving indomethacin (14.9+/-3.2 EE, n=7) and meloxicam (17.6+/-3.6 EE, n=11, 0.3 mg kg(-1)). Four piglets receiving meloxicam (0.3 mg kg(-1), i.v.) did not vomit during the delayed phase. The anti-emetic activity of two COX inhibitors suggests that prostaglandins contribute to the activation of the emetic reflex in response to cisplatin. PMID- 11897122 TI - Molecular characterization of a coccidian parasite cGMP dependent protein kinase. AB - The cGMP-dependent protein kinase (PKG) of Eimeria tenella and Toxoplasma gondii is the target of a novel coccidiostat that is effective against coccidiosis and toxoplasmosis in animal models. Preparations of native PKG enzyme from Toxoplasma and Eimeria contain a membrane-associated polypeptide (isoform-I) of about 110 kDa and a slightly smaller soluble polypeptide (isoform-II). Expression of T. gondii and E. tenella PKG cDNA clones in Toxoplasma yield similarly sized recombinant polypeptides, which co-migrate on SDS-polyacrylamide gels with the corresponding native isoforms. Results of targeted mutagenesis of potential translational initiation sites suggest that parasite isoform-II is a product of alternative translational initiation from an internal initiator methionine codon. Exclusive expression of isoform-II or isoform-I can be achieved by preventing initiation at the respective primary or secondary sites. Immunofluorescence analysis indicates that recombinant isoform-I localizes primarily to the parasite plasma membrane, while isoform-II remains cytosolic. Mutagenesis and metabolic labeling studies reveal that the observed membrane-association of full-length recombinant PKG is mediated by N-terminal myristoylation and palmitoylation at amino acids G2 and C4. We also confirm the functional significance of a putative third PKG allosteric site, common to apicomplexan PKGs but absent from vertebrate or insect PKGs. In assays with transiently transfected parasites, constructs harboring a mutation at this site express markedly lower levels of cGMP-dependent PKG activity, while a triple mutant bearing mutations in all three sites reduces kinase activity to background levels. PMID- 11897123 TI - Characterization of proteases involved in the processing of Plasmodium falciparum serine repeat antigen (SERA). AB - The Plasmodium falciparum serine repeat antigen (SERA), a malaria vaccine candidate, is processed into several fragments (P73, P47, P56, P50, and P18) at the late schizont stage prior to schizont rupture in the erythrocytic cycle of the parasite. We have established an in vitro cell-free system using a baculovirus-expressed recombinant SERA (bvSERA) that mimics the SERA processing that occurs in parasitized erythrocytes. SERA processing was mediated by parasite derived trans-acting proteases, but not an autocatalytic event. The processing activities appeared at late schizont stage. The proteases are membrane associated, correlating with the secretion and accumulation of SERA within the parasitophorous vacuole membrane (PVM). The activity responsible for the primary processing step of SERA to P47 and P73 was inhibited by serine protease inhibitor DFP. In contrast, the activity responsible for the conversion of P56 into P50 was inhibited by each of the cysteine protease inhibitors E-64, leupeptin and iodoacetoamide. Moreover, addition of DFP, E-64 or leupeptin to the cultures of schizont-stage parasites blocked schizont rupture and release of merozoites from PVM. These results indicate that SERA processing correlates to schizont rupture and the processing is mediated by at least three distinct proteases. PMID- 11897124 TI - Isolation of a Toxoplasma gondii cyclin by yeast two-hybrid interactive screen. AB - GAL4-based yeast two-hybrid cDNA libraries from Toxoplasma gondii RH strain were constructed and screened for interactors of a putative T. gondii cdc2-related kinase, TgCRK2. A screen of 3.2 million transformants yielded a single yeast clone that harbored a protein fusion capable of specifically interacting with TgCRK2. Sequencing revealed the cDNA insert (TgCYC1) had homology to the cyclin class of proteins. The TgCYC1 cDNA fragment was used to probe a conventional T. gondii cDNA library and a 2.65 kb cDNA coding for a predicted protein of 582 amino acids was obtained. Based on comparison with a 5'-RACE product from tachyzoite mRNA, the 2.65 kb cDNA for TgCYC1 appeared to be complete. TgCYC1 had the highest similarity to Plasmodium falciparum CYC1 and displayed sequence characteristics that place it in the cyclin H class of eukaryotic cyclins. In synchronous tachyzoite populations the level of TgCYC1 mRNA was unchanged indicating it is not cell cycle regulated at the mRNA level. TgCYC1 rescues the G(1)/S cyclin cell cycle defect in S. cerevisiae strain DL1 demonstrating that this apicomplexan cyclin can function in an established heterologous model system. PMID- 11897125 TI - A new expression vector for Crithidia fasciculata and Leishmania. AB - Crithidia fasciculata is a monogenetic parasite of insects. It grows in fully defined media without requiring serum, which facilitates biochemical analysis. We have constructed a series of expression systems that allows expression of transfected genes in the kinetoplastid protozoa Crithidia and Leishmania. These cells can be readily transfected with plasmid DNA by electroporation and transformants selected with various antibiotic resistance markers. 5'-Trans splicing signals and poorly defined regions within the 3'-untranslated regions of genes are required for optimal expression of genes in trypanosomatids. We, therefore, inserted the intergenic region of the C. fasciculata phosphoglycerate kinase (PGK) genes A and B, which allows polyadenylation of the target gene and spliced leader addition to the selectable marker gene. Part of the intergenic region of the PGK locus was added upstream of the target gene to permit its trans splicing. A 3'-untranslated sequence from the Crithidia glutathionylspermidine synthetase (GSPS) was also added to allow the polyadenylation of the selectable marker gene. Genes can be readily inserted using a multiple cloning site and can be expressed as a fusion protein with a poly-histidine sequence at either the N or C-terminus or fused with green fluorescent protein. Biologically active proteins can be expressed in C. fasciculata or L. amazonensis promastigotes and purified by affinity chromatography using a metal chelating column. PMID- 11897126 TI - A proton pumping pyrophosphatase in the Golgi apparatus and plasma membrane vesicles of Trypanosoma cruzi. AB - The proton pumping pyrophosphatase (H(+)-PPase) is an enzyme that has been identified in membranes of plant vacuoles, in the Golgi complex of plants and Chlamydomonas reinhardtii, and more recently in acidocalcisomes of different trypanosomatids and apicomplexan parasites. Immunofluorescence and immunoelectron microscopy studies using antibodies against the plant enzyme also suggested a plasma membrane localization in different stages of Trypanosoma cruzi. In this report we provide immunogold electron microscopy evidence of the presence of the H(+)-PPase in the Golgi complex and plasma membrane of epimastigotes of T. cruzi. Pyrophosphate promoted acidification of plasma membrane vesicles as determined using acridine orange. This activity was stimulated by K(+) ions, inhibited by the pyrophosphate analogs imidodiphosphate (IDP) and aminomethylenediphosphonate (AMDP) by KF, NaF and DCCD, and it had different responses to ions and inhibitors as compared with the activity present in acidocalcisomes. Surface localization of the H(+)-PPase was confirmed by experiments using biotinylation of cell surface proteins and immunoprecipitation with antibodies against H(+)-PPase. Taken together, these results are consistent with the presence of a functional H(+) PPase in the plasma membrane of these parasites. PMID- 11897127 TI - The Plasmodium vivax homologues of merozoite surface proteins 4 and 5 from Plasmodium falciparum are expressed at different locations in the merozoite. AB - Merozoite surface proteins of Plasmodium falciparum are one major group of antigens currently being investigated and tested as malaria vaccine candidates. Two recently described P. falciparum merozoite surface antigens, MSP4 and MSP5, are GPI-anchored proteins that each contain a single EGF-like domain and appear to have arisen by an ancient gene duplication event. The genes are found in tandem on chromosome 2 of P. falciparum and the syntenic region of the genome was identified in the rodent malarias P. chabaudi, P. yoelii and P. berghei. In these species, there is only a single gene, designated MSP4/5 encoding a single EGF like domain similar to the EGF-like domain in both PfMSP4 and PfMSP5. Immunization of mice with PyMSP4/5 provides mice with high levels of protection against lethal challenge with blood stage P. yoelii. In this study, we show that in P. vivax, which is quite phylogenetically distant from P. falciparum, both MSP4 and MSP5 homologues can be found with their relative arrangements with respect to the surrounding genes mostly preserved. However, the gene for MSP2, found between MSP5 and adenylosuccinate lyase (ASL) in P. falciparum, is absent from P. vivax. The PvMSP4 and PvMSP5 genes have a two-exon structure and encode proteins with potential signal and GPI anchor sequences and a single EGF-like domain near the carboxyl-terminus. Rabbit antisera raised against purified recombinant proteins show that each of the antisera react with distinct proteins of 62 kDa for PvMSP4 and 86 kDa for PvMSP5 in parasite lysates. Indirect immunofluorescence assays (IFA) localized PvMSP4 over the entire surface of P. vivax merozoites, as expected, whereas, the MSP5 homologue was found to be associated with an apical organellar location consistent with micronemes or over the polar prominence. PMID- 11897128 TI - Expression site activation in Trypanosoma brucei with three marked variant surface glycoprotein gene expression sites. AB - The genes for the Variant Surface Glycoprotein (VSG) of Trypanosoma brucei are transcribed in telomeric expression sites (ESs). There are about 20 different ESs per trypanosome nucleus. Usually, only one is active at a time, but trypanosomes can switch the ES that is active at a low rate (<10(-5) per cell per generation). To study activation and silencing of ESs, we have generated a line of T. brucei 427 with three ESs marked with a different drug resistance gene. We show that a selection with any combination of two of these drugs leads to an unstable double resistant phenotype in which the two ESs containing the corresponding marker genes switch backward and forward at a very high rate (>10(-1) per cell per generation). Unstable triple-resistant trypanosomes were not obtained. We conclude that the unstable rapid-switching state is a natural intermediate in ES switching. It only involves two ESs, whereas the other ESs are not expressed. Furthermore, we show that "inactive" ESs can exist at several different stable levels of activation. Whereas, a "silent" ES shows a low level of expression of promoter proximal sequences, the level of activation can be reversibly increased, leading to partially activated ESs. PMID- 11897129 TI - Biochemical and electron paramagnetic resonance study of the iron superoxide dismutase from Plasmodium falciparum. AB - Recombinant iron-containing superoxide dismutase (Fe-SOD) from Plasmodium falciparum was produced in a SOD-deficient strain of Escherichia coli, purified and characterised. The enzyme is a dimer, which contains 1.7 Fe equivalents and is sensitive to hydrogen peroxide (H(2)O(2)). Electron paramagnetic resonance (EPR) analysis showed two different signals, reflecting the presence of two different types of high-spin Fe sites with different symmetries. The role of the W71 residue during inactivation by H(2)O(2) of the P. falciparum Fe-SOD was studied by site-directed mutagenesis. First, the W71V mutation led to a change in the relative proportion of the two Fe-based EPR signals. Second, the mutant protein was almost as active as the wild-type (WT) protein but more sensitive to heat inactivation. Third, resistance to H(2)O(2) was only slightly increased indicating that W71 was marginally responsible for the sensitivity of Fe-SOD to H(2)O(2). A molecular model of the subunit was designed to assist in interpretation of the results. The fact that the parasite SOD does not belong to classes of SOD present in humans may provide a novel approach for the design of antimalarial drugs. PMID- 11897131 TI - Identification and biochemical characterisation of a protein phosphatase 5 homologue from Plasmodium falciparum. AB - We report the identification of a new serine/threonine phosphatase from Plasmodium falciparum at the DNA and protein levels. A 1.8 kb cDNA fragment encoding the protein phosphatase was identified via PCR amplification. The sequence has a coding capacity of 594 amino acids. Immunoblot analysis of P. falciparum extracts showed that antibodies generated against the His(6)-fusion protein recognise a protein of approximately 80 kDa. The deduced amino acid sequence shares 55% identity with a mouse protein, identified as Protein Phosphatase 5 (PP5). We show that the P. falciparum PP5 homologue (PfPP5) has all structural and functional characteristics of this class of enzymes. It contains three tetratricopeptide repeats (TPR) and a nuclear targeting sequence at its N terminus and a highly conserved C-terminal catalytic domain. Southern blot results are compatible with the existence of PfPP5 as a single copy gene. Purified recombinant protein, like the native protein enriched from P. falciparum extracts exhibited phosphatase activity that can be enhanced by both arachidonic and oleic acids, but not by myristic or stearic acid. In addition, the activity is inhibited by okadaic acid (OA) with an IC(50) of 4 nM. Immunofluorescence microscopy has localised PfPP5 preferentially to the nucleus. The function of PfPP5 is presently unclear, but like other PP5s of many eukaryotic organisms, it may have important regulatory functions in the parasite cell cycle. PMID- 11897130 TI - Characterization of a polymorphic Theileria annulata surface protein (TaSP) closely related to PIM of Theileria parva: implications for use in diagnostic tests and subunit vaccines. AB - Theileria annulata is a tick-transmitted protozoan that causes tropical theileriosis, an often fatal leukoproliferative disorder of cattle. To characterize and identify parasite proteins suitable as diagnostic antigens and/or vaccine candidates, a cDNA clone encoding a macroschizont stage protein was isolated and characterized (here designated TaSP). The gene, present as a single copy within the parasite genome, is transcribed in the sporozoite and schizont stage and codes for a protein of about 315 amino acids, having a predicted molecular weight of 36 kDa. Allelic variants were found within single parasite isolates and between isolates originating from different geographical regions. The N-terminal part contains a predicted signal peptide and the C terminal section encodes membrane-spanning regions. Comparison of a number of cDNA clones showed that both these sequence regions are conserved while the central region shows both size and amino acid sequence polymorphism. High identity of the N- and C-terminal regions with the polymorphic immunodominant molecule (PIM) of Theileria parva (identity of 93%), the existence of a central polymorphic region and two short introns within genomic clones suggest that the presented gene/protein may be the T. annulata homologue of PIM. However, the central region of TaSP has no significant identity with PIM, contains no repetitive peptide motifs and is shorter, resulting in a lower molecular weight. The existence of the predicted secretion signal peptide and membrane spanning regions suggest that TaSP is located at the parasite membrane. PMID- 11897132 TI - Trypanosoma brucei expression-site-associated-gene-8 protein interacts with a Pumilio family protein. AB - The expression site (ES) loci of Trypanosoma brucei are a valuable model for allelic exclusion and post-transcriptional regulation in a highly divergent eukaryote. ES exist to facilitate the expression and switching of the variant surface glycoproteins (VSG) that are central to trypanosome virulence and persistence. A collection of other potential virulence determinants, known as expression-site-associated-genes (ESAGs), are co-transcribed from the single upstream promoter. ESAGs may be involved in regulating the transcriptional state of the ES, as well as contributing additional surface proteins and receptors. We have previously shown that a putative regulatory protein, ESAG8, accumulates within the nucleolus, although 20% of the protein is cytoplasmic. Here we identify TbPUF1, a cytoplasmic ESAG8-interacting protein that falls into the Puf family of regulators of mRNA stability. Our experiments show that, as in other Puf family proteins, the most C-terminal repeats of TbPUF1 mediate its interaction with ESAG8. TbPUF1 is essential for cell viability, and preliminary results suggest that its overexpression seriously affects parasite virulence. T. brucei is the most evolutionary divergent organism in which a Puf family protein has been identified, and our initial experiments suggest that this protein may also regulate RNA stability in trypanosomes. PMID- 11897133 TI - Amino-terminal control of transgenic protein expression levels in Toxoplasma gondii. AB - Comparing the steady-state expression levels of recombinant proteins in Toxoplasma gondii parasites indicates considerable variability, and this has sometimes caused difficulties in the engineering of transgenic parasites. Anecdotal observations suggested that alteration of the N-terminus, e.g. by engineering as a fusion protein, permits stable expression of various transgenes that were previously difficult to express in their native form. We have exploited the sensitivity and quantitative nature of fire-fly luciferase (LUC) to examine expression levels in further detail. Fusing the 26 N-terminal residues derived from chloramphenicol acetyl transferase (DeltaCAT) to LUC permits efficient transient or stable luciferase expression in transgenic parasite tachyzoites, providing a useful reporter for studies in T. gondii. Site-directed mutagenesis was used to alter the second codon of DeltaCAT-LUC to encode all 20 possible amino acids, and these constructs showed that changes in the second amino acid can have dramatic effects on luciferase activity, with Ala, Glu, and Asp codons yielding the highest expression levels. Similar results were observed for the expression of both GFP and the T. gondii HXGPRT gene, demonstrating the generality of this effect. PMID- 11897134 TI - A developmentally regulated metalloprotease secreted by host-stimulated Ancylostoma caninum third-stage infective larvae is a member of the astacin family of proteases. PMID- 11897135 TI - Mutation in position 167 of isotype 1 beta-tubulin gene of Trichostrongylid nematodes: role in benzimidazole resistance? PMID- 11897136 TI - Limited polymorphism of the vaccine candidate merozoite surface protein 4 of Plasmodium falciparum. PMID- 11897137 TI - Comparative study of Trypanosoma rangeli and Trypanosoma cruzi telomeres. PMID- 11897138 TI - Tetracycline-regulated RNA interference in Trypanosoma congolense. PMID- 11897139 TI - Nucleoside hydrolase from Leishmania (L.) donovani is an antigen diagnostic for visceral leishmaniasis. PMID- 11897140 TI - Definition of the minimal domain of CIDR1alpha of Plasmodium falciparum PfEMP1 for binding CD36. PMID- 11897144 TI - Poly(ethylene oxide)-block-poly(L-amino acid) micelles for drug delivery. AB - Block copolymer micelles encapsulate water insoluble drugs by chemical and physical means, and they may target therapeutics to their site of action in a passive or active way. In this review, we focus on micelles self-assembled from poly(ethylene oxide)-block-poly(L-amino acid) (PEO-b-PLAA). A common theme in these studies is the chemical modification of the core-forming PLAA block used to adjust and optimize the properties of PEO-b-PLAA micelles for drug delivery. Micelle-forming block copolymer-drug conjugates, micellar nanocontainers and polyion complex micelles have been obtained that mimic functional aspects of biological carriers, namely, lipoproteins and viruses. PEO-b-PLAA micelles may be advantageous in terms of safety, stability, and scale-up. PMID- 11897145 TI - Polyether-polyester diblock copolymers for the preparation of paclitaxel loaded polymeric micelle formulations. AB - A number of hypersensitivity reactions have been attributed to the presence of Cremophor((R)) EL in the current formulation for paclitaxel. This has led to the development of formulations for paclitaxel employing polyether-polyester diblock copolymers as micelle forming carriers. Diblock copolymers of methoxypolyethylene glycol-block-poly(D,L-lactide) (MePEG:PDLLA) were synthesized from monomers of D,L-lactide and MePEG by a ring opening bulk polymerization in the presence of stannous octoate. Up to 25% paclitaxel could be loaded into matrices of MePEG:PDLLA (60:40, MePEG molecular weight of 2000) using the solution casting method. Dissolution of paclitaxel/copolymer matrices in aqueous media resulted in complete solubilization of paclitaxel within the hydrophobic PDLLA core of the micelles. This review article describes the synthetic reaction conditions influencing the degree of conversion of monomer to copolymer, thermal properties, critical micelle concentrations of copolymers, methods of incorporation of paclitaxel into copolymer matrices and subsequent constitution in aqueous media and biological evaluations of micellar paclitaxel. PMID- 11897146 TI - Block copolymer micelles for delivery of gene and related compounds. AB - Block copolymers composed of a cationic segment and a hydrophilic segment spontaneously associate with polyanionic DNA to form block copolymer micelles. The distinct feature of the associate is that the core of the polyion complex between DNA and the polycation is coated by a layer of the hydrophilic polymer. The characteristic core-shell structure endows the associate with a high colloidal stability and reduced interaction with blood components. These desirable properties are the major advantages of the micellar DNA delivery system for in vivo application. In this article, the synthesis of block copolymers as well as graft copolymers utilized as DNA delivery systems are described. Particular emphasis is devoted to the association behavior and the physicochemical properties of polyion complex micelles entrapping DNA and related substances in relation to the biological aspects of the associates. Biodistribution and the factors that affect the intracellular fate of the micelles is also addressed based on recent studies in this field. PMID- 11897147 TI - Pluronic block copolymers: novel functional molecules for gene therapy. AB - Pluronic block copolymers are recognized pharmaceutical excipients listed in the US and British Pharmacopoeia. They have been used extensively in a variety of pharmaceutical formulations including delivery of low molecular mass drugs and polypeptides. This review describes novel applications of Pluronic block copolymers in gene therapy. In particular, these molecules can modify the biological response during gene therapy in the skeletal muscle, resulting in an enhancement of the transgene expression as well as an enhancement of the therapeutic effect of the transgene. Furthermore, Pluronic block copolymers are versatile molecules that can be used as structural elements of the polycation based gene delivery systems (polyplexes). Based on these studies, the use of block copolymers in gene delivery is a promising area of research, in which new and important developments are expected. PMID- 11897148 TI - PEG-based micelles as carriers of contrast agents for different imaging modalities. AB - The review deals with the fast growing field of diagnostic micelles. The need and requirements for microparticulate contrast agents are discussed. Brief analysis of the micellization process and micelle properties shows that micelles made of amphiphilic co-polymers seem to be the most attractive for practical application. These micelles can be prepared from the variety of co-polymers including hydrophilic polymers grafted on one terminus with lipid residues. Polymeric micelles are considered loaded or modified with various contrast reporter moieties for gamma-scintigraphy, magnetic resonance imaging (MRI), and computed tomography (CT). Their in vitro and in vivo properties are discussed and the results of the initial animal experiments are presented. Mixed micelles were prepared from diacyllipid-polyethylene glycol (PEG) conjugates and polymeric amphiphilic chelates, containing entrapped metals, such as 111-In or Gd, and used for the experimental gamma- and MR imaging of various components of lymphatic system in rabbits. The method is also described to prepare polymeric iodine containing PEG-based micelles which may act as a long-circulating blood pool imaging agent for CT. Experimental CT-imaging performed in mice and rabbits demonstrated high potential of a micellar contrast agent. PMID- 11897149 TI - Activity coefficients of antibiotics in aqueous NaCl solutions at 298.2 K. AB - Data at 298.2 K for aqueous NaCl solutions containing either D-phenylglycin, D-(p hydroxy)phenylglycine, 6-amino penicillanic acid (6-APA), amoxicillin or ampicillin, are reported. The mean ionic activity coefficients of NaCl were determined from measurements of the responses of a sodium and a chloride ion selective electrode. The activity coefficients of the precursors or the antibiotics were calculated from the values of the mean ionic activity coefficients using the exact cross differential relation between them. The correlation of solubility data using the activity coefficients measured in this work shows the same puzzling results previously observed in systems containing amino acids. PMID- 11897150 TI - Proteolysis of protein C in pooled normal plasma and purified protein C by activated protein C (APC). AB - Protein C is a vitamin-K dependent zymogen of the anti-coagulant serine protease activated protein C (APC). In this paper, we report four lines of evidence that APC can activate protein C in pooled normal plasma, and purified protein C. First, the addition of APC to protein C-deficient plasma supplemented with protein C produces a prolongation of the clotting time of plasma that is proportional to the amount of protein C. This behavior was observed with APC from the Chromogenix APC resistance kit (Dia Pharm, Franklin, OH, USA) and from APC derived from the thrombin activation of human protein C (Enzyme Research Laboratories, South Bend, IN, USA). Secondly, using immunoblotting after gel electrophoresis, the disappearance of epitopes for monoclonal antibodies that recognize protein C but not APC indicates a time course for the activation by APC of protein C in pooled normal plasma and protein C purified from plasma. Thirdly, the same time course for the disappearance of protein C specific epitope can be followed using ELISA. Finally, protein C can be activated by APC as indicated by the increase in APC specific synthetic substrate Tryp-Arg-Arg-p nitroaniline hydrolysis. Kinetic data indicate a value of 4.7+/-0.4 mM(-1) s(-1) for the activation of protein C by APC under physiological conditions and in the presence of calcium. These observations document that APC must function not only in the inactivation of activated factors V and VIII, but also in the activation of protein C. This additional action of APC may be important to consider more broadly because of APC in the treatment of sepsis. PMID- 11897151 TI - Molecular modelling of amphotericin B-ergosterol primary complex in water. AB - The properties of amphotericin B-ergosterol primary complex have been studied with the use of the molecular dynamics simulation. Possible geometries of the complex were tested first in order to find the structures with the most favourable values of the intermolecular interactions energy. The molecules studied possessed a tendency to fit each other's shapes, which favours intermolecular van der Waals interactions. The main simulations were performed for the best structures found. Presence of hydrogen bonds between the sterol hydroxyl group and polar fragments of mycosamine (most frequently 2'OH) was coupled with a relatively high level of the intermolecular energy values. The structures obtained are hardly comparable to the hypothetical and 'computational' models of the antibiotic-sterol complex. The geometries found are not suitable to assemble the presupposed structure of the water channel, however, the existence of the complex in the shape anticipated is not in contradiction to the results of biophysical experiments on the complexation in water and in hydroalcoholic media. PMID- 11897153 TI - Different effects of propofol and nitrosopropofol on DMPC multilamellar liposomes. AB - The mechanisms of reaction of propofol with nitrosoglutathione lead to the formation of an active species which was identified, and then synthesised, as 2,6 diisopropyl-4-nitrosophenol. In the present work, we demonstrate the in vitro formation of 2,6-diisopropyl-4-nitrosophenol, then we discuss the interaction of propofol and 2,6-diisopropyl-4-nitrosophenol with dimyristoylphosphatidylcholine and egg yolk phosphatidylcholine multilamellar liposomes using differential scanning calorimetry and spin labelling techniques. It was demonstrated that both molecules are highly lipophylic and absorb almost entirely in the lipid phase. The thermotropic profiles showed that these molecules affect the temperature and the co-operativity of the gel-to-fluid state transition of the liposomes differently: the effects of 2,6-diisopropylphenol on the lipid organisation are quite similar to phenol and coherently interpretable in terms of the disorder produced in the membrane by a bulky group; 2,6-diisopropyl-4-nitrosophenol is a stronger perturbing agent, and ESR spectra suggest that this is due to a relative accumulation of the molecule into the interfacial region of the bilayer. PMID- 11897152 TI - The application of fluorescence correlation spectroscopy in detecting DNA condensation. AB - We report the application of fluorescence correlation spectroscopy (FCS) in characterizing conformational changes (condensation) of chemically well-defined DNA plasmids. The plasmids: pHbetaAPr-1-neo (10 kbp, contour length 3.4 microm) and pBluescript SKt (2.96 kbp, contour length 1.02 microm) were imaged by a confocal fluorescence microscope using two fluorescent probes: ethidium bromide (EtBr) and propidium iodide (PrIo). It became clear that the DNA molecule exhibits discrete conformational change between the coil and globule states with the addition of a small amount (the order of magnitude being 10(-5) M) of cationic surfactant, spermine and hexadecyltrimethyl ammonium bromide (HTAB). When the concentrations of both condensing agents are smaller than 6.0x10(-6) M and 2.0 x 10(-6) M for the 10 and 2.96 kbp, both plasmids are in the extended coil state with diffusion constants D(10 kbp)=9.6 x 0(-13) m(2) s(-1) and D(2.96 kbp)=2.5x10(-12) m(2) s(-1), respectively. When the condensing agent in a concentration higher than 1.10 x 10(-5) M is added to pHbetaAPr-1-neo (10 kbp), plasmids are in the condensed globular state and their diffusion constants are D(10 kbp)=8.0 x 10(-12) m(2) s(-1) (spermine) and D(10 kbp)=5.5x10(-12) m(2) s( 1) (HTAB). The globular state of the pBluescript SKt (2.96 kbp) plasmids is characterized by diffusion constants equal to D(2.96 kbp)=9.2x10(-12) m(2) s(-1) (spermine) and D(2.96 kbp)=8.2x10(-12) m(2) s(-1) (HTAB). PMID- 11897154 TI - Flunitrazepam-membrane non-specific binding and unbinding: two pathways with different energy barriers. AB - The effect of molecular packing on flunitrazepam's ability to interact with bio membranes was studied using dipalmitoylphosphatidylcholine monomolecular layers at the air-water interface as a model membrane. Flunitrazepam penetrated from the subphase into monolayers at lateral pressures below 44.8 mN/m and induced their concentration-dependent expansion. As inferred from the values of compressibility modulus, the elasticity of the liquid-condensed phase decreased in the presence of flunitrazepam. Although this drug hardly penetrated into high-packed monolayers, it was easily incorporated in the low-packed ones at an extent sufficient to reach the partition equilibrium. Below a molecular area of 75 A(2), contrary to what would be expected, the drug surface concentration increased as a function of surface pressure, suggesting that after its penetration in disordered phases, it became energetically or physically trapped in newly-formed liquid condensed clusters. The phenomenon of flunitrazepam penetration and release would have different energy barriers depending on the membrane phase-state. PMID- 11897155 TI - Molecular characterization of a ligand-tethered parathyroid hormone receptor. AB - It was recently shown that the covalent tethering of the N-terminus of parathyroid hormone (PTH) to the seventh helical bundle of the G-protein coupled PTH-receptor (PTH1R) leads to autoactivation [Shimizu et al., J. Biol. Chem. 275 (2000) 19456-19460]. Here, we have developed molecular models for the interaction of PTH(1-11) tethered to PTH1R and refined them with molecular dynamics simulations. The starting structure of the ligand/receptor complex is based on experimental data from a series of spectroscopic structural studies of PTH(1-34) and the extracellular domains of PTH1R and intermolecular contact points derived from photoaffinity labeling. The resulting PTH1R/[Arg(11)]PTH(1-11) complex has the N-terminus of PTH interacting with residues of the third extracellular loop of PTH1R, as a possible mode for receptor activation. The hydrophobic residues leucine-5 and methionine-8, centrally located in the N-terminal alpha-helix of PTH(1-11), are located in deep, well-defined hydrophobic pockets in the central core of the seventh helical bundle, consistent with the requirement of these amino acids for autoactivation. We postulate that the improved signaling properties of [Arg(11)]PTH(1-11) over wild type PTH(1-11) is due to a stable hydrogen bond between Arg(11) and E444, at the beginning of TM7. The model provides atomic insight into currently available biochemical data as well as numerous putative ligand/receptor interactions, and thereby may further the rational design of reduced-size PTH agonists at the PTH1 receptor. PMID- 11897156 TI - Interfacial tension of phosphatidylcholine-cholesterol system in monolayers at the air/water interface. AB - Interfacial tension of an egg lecithin-cholesterol system was measured across the whole concentration range. Surface pressure-area isotherm measurements were carried out in a Langmuir trough at the air/water interface at room temperature (22 degrees C). The interfacial tension of the air/water interface was divided into contributions of components. The interfacial tension of a 1:1 complex between phosphatidylcholine and cholesterol was calculated. Its value equals 18 mN/m. The difference between the stability constant of 1:1 complex in the bilayer and the monolayer at the air/water interface is discussed. PMID- 11897157 TI - Nicotinamide and ketamine reduce infarct volume and DNA fragmentation in rats after brain ischemia and reperfusion. AB - The possible ability of nicotinamide and ketamine to decrease infarction volume and DNA fragmentation was investigated in a middle cerebral artery occlusion rat model. DNA fragmentation was measured with an enzyme linked immunoassay. Control infarct volume was 223.8 +/- 10.6 mm(3). Ketamine alone did not alter infarct volume, 233.2 +/- 61.8 mm(3). Nicotinamide alone did not alter infarct volume, 235.2 +/- 62.8 mm(3). The combination of ketamine and nicotinamide decreased infarct volume to 83.8 +/- 35.2 mm(3). Ketamine produced hypothermia. Nicotinamide and ketamine decreased brain swelling and DNA fragmentation in the cerebral cortex, striatum and hippocampus in rats perfused for 6 or 24 h. Ketamine may synergize the actions of nicotinamide and partially prevent brain damage from ischemia and reperfusion. PMID- 11897158 TI - Auditory cortical activation in Finnish and Swedish speaking Finns: a magnetoencephalographic study. AB - To study the effects of linguistic background on auditory processing, magnetoencephalographic responses for pure tones (120 Hz, 1 and 4 kHz), [u] and a complex tone (with three pure tone components corresponding to the three lowest formant frequencies of [u]) were recorded in ten Finnish and ten Swedish speaking Finnish males. Auditory cortical activation, maximal at about 100 ms after stimulus onset, was stronger in the right hemisphere (RH) for all stimuli. At 175 225 ms, Swedish speaking subjects had larger inter-hemispheric differences and different signal morphology in the RH than Finnish speaking subjects, suggesting that linguistic background influences basic auditory processes. Possibly, Swedish speaking subjects had retained a juvenile response component due to their bilingual surrounding after early childhood. PMID- 11897159 TI - Menthol: a natural analgesic compound. AB - Menthol, after topical application, causes a feeling of coolness due to stimulation of 'cold' receptors by inhibiting Ca++ currents of neuronal membranes. Since Ca++ channel blockers are endowed with analgesic properties, the aim of the present study was to investigate the potential antinociceptive effect of menthol. (-)-Menthol produced a dose-dependent increase in the pain threshold in the mouse hot-plate (3-10 mg kg(-1) p.o.) and abdominal constriction (3-10 mg kg(-1) p.o.; 10 microg per mouse intracerebroventricularly (i.c.v.)) tests. The antinociceptive effect of (-)-menthol was antagonised by the unselective opioid antagonist naloxone and by the selective kappa-antagonist nor-NBI. Conversely, CTOP (mu-antagonist), 7-benzylidenenal-trexone (delta(1) antagonist) and naltriben (delta(2) antagonist) did not prevent (-)-menthol antinociception. In both tests, (+)-menthol (10-50 mg kg(-1) p.o.; 10-30 microg per mouse i.c.v.) was unable to modify the pain threshold. These results indicate that (-)-menthol is endowed with analgesic properties mediated through a selective activation of kappa-opioid receptors. PMID- 11897160 TI - Seizure-mediated accumulation of the beta subunit of Ca2+/calmodulin-dependent protein kinase II in nuclei of mouse brain cells. AB - We identified a 45-kDa protein by 2D electrophoresis that was enhanced following pentylenetetrazol (PTZ)-mediated seizures. Mass-spectrography of this protein revealed the beta subunit of Ca2+/calmodulin-dependent protein kinase II (CaMKIIbeta), although no evidence for increase in bulk CaMKIIbeta transcripts was obtained. Physicochemical parameters of the 45-kDa species coincided with those of the type 7 isoform of CaMKIIbeta, CaMKIIbeta7. Reverse transcription polymerase chain reaction revealed the existence of the CaMKIIbeta7 transcript in the mouse brain, but its RNA content was small and was not elevated by PTZ injection. CaMKIIbeta7 protein is thought to be accumulated in the nuclei of brain cells by PTZ-mediated seizure via some cellular mechanisms other than transcriptional and post-transcriptional regulation. PMID- 11897161 TI - A mouse model for peripheral neuropathy produced by a partial injury of the nerve supplying the tail. AB - We attempted to develop a mouse model for peripheral neuropathy by a partial injury of the nerve supplying the tail. Under enflurane anesthesia, the unilateral superior caudal trunk was resected between the S3 and S4 spinal nerves. Tests for thermal allodynia were conducted by immersing the tail into 4 or 38 degrees C water. The mechanical allodynia was assessed by stimulating the tail with a von Frey hair (1.96 mN, 0.2 g). After the nerve injury, the experimental animals had shorter tail withdrawal latencies to cold and warm water immersion than the presurgical latency, and exhibited an increase in tail response to von Frey stimulation. We interpret these abnormal sensitivities as the signs of mechanical, cold and warm allodynia following the superior caudal trunk injury in the mouse. PMID- 11897162 TI - Repetitive concentric wave-ring spread of oligemia/hyperemia in the sensorimotor cortex accompanying K(+)-induced spreading depression in rats and cats. AB - Vascular changes accompanying spreading depression (SD) remain controversial. We examined dynamic alterations of local cerebral blood volume (CBV) during SD by observing light transmission at an isosbestic point of hemoglobin (550 nm) in seven rats and five cats under alpha-chloralose/urethane anesthesia. The two species were used for comparison between the lissencephalic and gyrencephalic brains. We found that a concentrated K(+) solution microinjected into the sensorimotor cortex provoked CBV changes that appeared as a repetitive propagation of concentric wave-rings of ischemia followed by hyperemia expanding peripherally from the injection site at speeds of 1.9-3.2 mm/min. The dynamic CBV changes continued repeatedly every 1-5 min for more than 30 min in three rats, ceased within 30 min in three rats and remained at the site of K(+) injection in one rat. Similar repeated CBV changes occurred in two out of five cats. PMID- 11897163 TI - Intrathecal alpha2 adrenoceptor agonist clonidine inhibits mechanical transmission in mouse spinal cord via activation of muscarinic M1 receptors. AB - We examined the role of the spinal muscarinic receptor subtype in the anti nociceptive effect of intrathecal (i.t.) alpha2 adrenoceptor agonist clonidine in mice. I.t. injection of the muscarinic receptor antagonist atropine completely inhibited i.t. clonidine-induced increase in the mechanical threshold, but did not affect the increase in tail-flick latency induced by i.t. clonidine. The clonidine-induced increase in mechanical threshold was inhibited by i.t. injection of the M1 receptor antagonist pirenzepine in a dose-dependent manner, and by the M3 receptor antagonist 4-DAMP, but not by the M2 receptor antagonist methoctramine. The potency of pirenzepine was greater than that of 4-DAMP. These results suggest that the clonidine-induced increase in mechanical threshold is mediated via the activation of M1 receptors in the spinal cord. PMID- 11897164 TI - Amphetamine inhibits behavior-related neuronal responses in substantia nigra pars reticulata of rats working for sucrose reinforcement. AB - Changes in activity of basal ganglia neurons, especially those in the striatum, are thought to underlie the characteristic behavioral patterns produced by d amphetamine (AMPH). To study the role of the substantia nigra pars reticulata (SNr), a major basal ganglia output nucleus, we recorded from SNr neurons before and after a behaviorally activating dose of AMPH (0.5 mg/kg) in rats trained to nosepoke for sucrose reinforcement. Before AMPH, task-related behaviors were associated primarily with increases or both increases and decreases in SNr firing. Although these same behavior-related patterns persisted after AMPH, their relative magnitude was significantly attenuated. Units unresponsive during task events were unaffected by AMPH. Thus, rather than change the overall level of SNr firing, a behaviorally active dose of AMPH exerts context-dependent effects on the activity of SNr neurons. PMID- 11897165 TI - Dissociation of the associative and visceral sensory components of taste aversion learning by tetrodotoxin inactivation of the parabrachial nucleus in rats. AB - The parabrachial nucleus (PBN) has been proposed as the associative site for conditioned taste aversion. Previous evidence has shown that functional blockade of the PBN by tetrodotoxin (TTX) produces retrograde disruption of lithium induced taste aversions in rats. However, given the PBN role in processing visceral cues and the long duration of the lithium-induced aversive effects, an interpretation based on lithium chloride processing deficits can not be ruled out. The aim of the present study was to use the unconditioned stimulus (US) pre exposure phenomenon to explore the effect of PBN inactivation by intracerebral TTX microinjections on visceral processing. Three intraperitoneal (i.p.) lithium chloride injections (0.15 M; 2% b.w.) applied before the conditioning session, but not isotonic saline i.p. injections, interfered with the acquisition of a learned aversion to a cider vinegar solution (3%) in cannulated control rats. Bilateral PBN inactivation by TTX (10 ng) applied immediately after each LiCl injections disrupted the US pre-exposure effect, thus confirming its sensory role. However, PBN inactivation 30 min after LiCl injections did not interfere with the US pre-exposure effect, in spite of the fact that an identically timed PBN blockade after the acquisition trial disrupted the acquisition of taste aversions. These results stand for the associative role of PBN in taste aversion learning induced by lithium chloride, independent of its sensory role. It is concluded that PBN activity is required after the conditioning trial for the taste-visceral association to take place. PMID- 11897166 TI - Corticopontocerebellar pathway from the prearcuate region to hemispheric lobule VII of the cerebellum: an anterograde and retrograde tracing study in the monkey. AB - The relationship between axons derived from the prearcuate region and pontine neurons projecting to contralateral cerebellar hemispheric lobule VII, lobulus petrosus (LP) of the paraflocculus or the dorsal paraflocculus (DPFl) was investigated in the monkey. The frontopontine axons were labeled with biotinylated dextran and pontocerebellar neurons with cholera toxin subunit B or fast blue. Labeled frontopontine axons and terminals were seen in the dorsal, medial, paramedian and dorsolateral parts of the pontine nuclei. The distribution of the labeled frontopontine axons overlapped that of labeled neurons projecting to hemispheric lobule VII but did not overlap that of labeled neurons projecting to LP or DPFl. The pathway from the prearcuate region to hemispheric lobule VII may provide an anatomical substrate for the involvement of hemispheric lobule VII in voluntary eye movements. PMID- 11897167 TI - Bullfrog retinal bipolar cells may express heterogeneous glycine receptors at dendrites and axon terminals. AB - Subcellular localization and properties of glycine receptors on bipolar cells (BCs) were studied using whole-cell recordings and non-stationary noise analysis (NSNA) in bullfrog retinal slices. The currents elicited by focally applied glycine were of comparable amplitudes at the dendrites and axon terminals of both OFF and ON BCs. Moreover, glycine receptors were also expressed at the axons of some BCs. NSNA revealed that the weighted mean single-channel conductance of the glycine receptors at the dendrites (18.2 pS) was significantly larger than that of those at the axon terminals (8.1 pS), thus implying that the glycine receptors on bullfrog retinal BCs may be heterogeneous at these two sites. PMID- 11897168 TI - Mental navigation in humans is processed in the anterior bank of the parieto occipital sulcus. AB - We examined the brain regions which were activated during mental navigation; functional magnetic resonance imaging was performed in 16 right-handed male volunteers. The anterior bank of the parieto-occipital sulcus (APO) was strongly activated in all 16 subjects examined. In group study, the retrosplenial area, the bilateral angular gyrus/occipital cortex junction, the left superior premotor area, the right parahippocampal gyrus, and the right cerebellum were activated commonly across 16 subjects. The APO region activated during mental navigation appeared to be equivalent to the visual area V6A in monkeys and to subserve egocentric spatial processes. PMID- 11897169 TI - Hypoxia/reoxygenation induces cell injury via different mechanisms in cultured rat cortical neurons and glial cells. AB - Hypoxia/reoxygenation (H/R) causes cell injury/death. We examined the protection by drugs intervening at various stages of the injury cascade in cultured neurons and glia. Primary cultures of rat cortical neurons and mixed glia were subjected to H/R. Measurements of cell death (by lactate dehydrogenase release into the medium) and viability (by MTT reduction) indicated that H/R led to time-dependent injury in both neuronal and mixed glial cultures. The extent of cell injury in neurons was significantly greater than in glia cells. Pretreatment with (+)-MK 801 hydrogen maleate (MK-801) (an N-methyl-D-aspartate antagonist), N(omega) nitro-L-arginine methyl ester (L-NAME) (an inhibitor of nitric oxide synthase) or free radical scavengers reduced the extent of the H/R-elicited neuronal damage. MK-801, in contrast, was without effect on glial cells while L-NAME was effective. Our results suggest differential mechanism(s) and susceptibility to injury caused by H/R for neurons and mixed glia. PMID- 11897170 TI - No association between the lipoprotein lipase S447X polymorphism and Alzheimer's disease. AB - Results from various genetic association studies of the lipoprotein lipase (LPL) S447X polymorphism and Alzheimer's disease (AD), range from a statistically significant negative association of clinically examined patients to a non significant but consistent trend for under-representation of the X447 allele in neuropathologically confirmed subjects. In this report we have compared the distribution of the above polymorphism in an independent group of clinically diagnosed AD patients, including a subgroup where the disease was pathologically confirmed, and a spousal control group. No statistically significant differences emerged from this comparison. We conclude that LPL cannot be a major factor in pathogenesis of AD. PMID- 11897171 TI - Demographic differences in patterns in the incidence of smoking cessation: United States 1950-1990. AB - PURPOSE: Current measures of successful quitting are insensitive to changes induced by tobacco control activities. We evaluated whether changes in the incidence of successful quitting, a new measure of cessation, can inform policy makers how population subgroups responded. METHODS: Smokers from National Health Interview Surveys (NHIS) (1965 through 1992, n = 140,199) were used to determine the number of current smokers eligible to quit at the beginning of each year from 1950 through 1990. Incidence of quitting, computed for different demographic subgroups, was the ratio of those newly successfully quit each year to those eligible to quit. RESULTS: Overall, incidence increased over fivefold, from < 1% in 1950 to a still low 5% in 1990. When the health risks of smoking were first disseminated, middle-aged men had the highest quitting incidence. Gender differences in younger smokers occurred following the beginning of the public health campaign of the mid 1960s, as the dangers of smoking to the fetus were documented. Younger adult smokers appeared to increase quitting markedly in the 1970s, around the beginning of the nonsmokers' rights movement. Quitting patterns in middle-aged African Americans were similar to whites, although at much reduced levels. Younger African Americans had low quitting incidence until 1989. Incidence differed by educational attainment; regardless of age, during the 1970s and 1980s, those with some college increased their quitting incidence markedly. CONCLUSION: Incidence of quitting is a sensitive indicator of relatively short term changes in successful quitting in population subgroups and should facilitate evaluation efforts. PMID- 11897172 TI - Interaction and intervention modeling: predicting and extrapolating the impact of multiple interventions. AB - PURPOSE: Methods called interaction and intervention modeling are presented. Interaction modeling examines the interactions between variables as the basis for predicting the impact of multiple variables on a target population and on populations with difference distributions of risk factors. Intervention modeling incorporates these interactions and aims to extrapolate the impact of multiple interventions to new populations. The aim is to develop methods that will be useful for modeling and comparing intervention strategies using existing data and standard statistical methods. METHODS: Traditional hypothesis testing methods used for randomized clinical trials and cohort studies and extrapolating the results to new populations are compared with interaction and intervention modeling methods. Interaction and intervention modeling utilizes the same data as the traditional approach but examines the impact of multiple simultaneous interactions and allows extrapolation of the results to populations with different prevalences and distributions of risk factors. An example using real data demonstrates the potential of interaction and intervention modeling to predict the impact of multiple interacting variables and to compare the impact of alternative interventions. RESULTS: The methods outlined take into account the impact of the magnitude of the relative risks, prevalence of risk factors, and interaction of risk variables when predicting the impact on a new population or extrapolating the results of one or more interventions on a new population. Traditional methods that do not take into account interactions are shown to produce different conclusions from the intervention modeling approach that incorporates interactions. The impact of the intervention modeling approach compared with the traditional approach will be quite variable depending on the prevalence of the risk factors and their extent of interaction. CONCLUSIONS: Studies designed to test a hypothesis treat most variables as potential confounding variables adjusting for their impact and their interactions as part of the analysis using traditional regression methods. Interaction and intervention modeling focuses on the interactions themselves and allows comparison of the effectiveness of alternative interventions. PMID- 11897173 TI - Green tea consumption and serum lipids and lipoproteins in a population of healthy workers in Japan. AB - PURPOSE: To examine the relation between green tea consumption and serum lipids and lipoproteins. METHODS: The subjects were 13,916 workers (8476 men and 5440 women) aged 40-69 years at over 1000 workplaces in Nagano prefecture, central Japan. They underwent health screening offered by a single medical institute between April 1995 and March 1996 and did not have morbid conditions affecting serum cholesterol levels. Serum concentrations of total cholesterol, high-density lipoprotein (HDL) cholesterol and triglycerides were measured at the screening. The consumption of green tea and other life-style characteristics were ascertained by a questionnaire. The data were analyzed with multivariate linear model. RESULTS: Daily consumption of green tea was reported by 86.7% of subjects. Green tea consumption was, statistically, significantly associated with lower levels of serum total cholesterol in both men and women while its associations with serum triglycerides and HDL cholesterol were not statistically significant. The inverse association of serum total cholesterol with green tea consumption appeared to level off at the consumption of more than 10 cups/day. Excluding the outlying subjects drinking more than 10 cups/day (0.4%), the regression analysis adjusting for age, body mass index, ethanol intake, smoking habit, coffee intake, and type of work showed that daily consumption of one cup of green tea was associated with a reduction in serum total cholesterol by 0.015 mmol/L (95% confidence interval 0.006 to 0.024, p < 0.001) in men and 0.015 mmol/L (0.004 to 0.025, p < 0.01) in women. After additional adjustment for selected dietary factors, the inverse association remained statistically significant; one cup of green tea per day was associated with a reduction in serum total cholesterol by 0.010 mmol/L (0.001 to 0.019, p = 0.03) in men and 0.012 mmol/L (0.001 to 0.022, p = 0.03) in women. CONCLUSION: Consumption of green tea was associated with lower serum concentration of total cholesterol in Japanese healthy workers age 40 69 years; however, green tea consumption was unrelated to serum HDL-cholesterol and triglycerides. PMID- 11897174 TI - Detection of pulmonary tuberculosis in Chiapas, Mexico. AB - PURPOSE: To estimate the prevalence of undiagnosed pulmonary tuberculosis (PTB) and the sensitivity of bacilloscopy in the border region of Chiapas, Mexico. METHODS: We actively sought individuals aged 15 years or more with chronic cough from the Border Region of Chiapas, Mexico in three settings: one regional hospital, seven Primary Care Centers (PCC), and 32 communities. Individuals (a total of 899) reporting chronic cough were asked to provide three samples of sputum for acid-fast smears and cultures. The quality of acid-fast smears was evaluated using culture as the gold standard. RESULTS: We obtained sputum specimens from 590 of 899 individuals with chronic cough. A diagnosis of PTB was confirmed in 78. A conservative estimate of the overall prevalence of PTB at the population level was 151 per 100,000 (95% CI: 88 to 241). In the regional hospital, the estimated case detection rate was 66% (29/44). The proportion of candidates for PTB therapy that were actually on treatment was 50% (14/28) at the PCC and 11% (2/19) in the communities. The sensitivity of the bacilloscopy was about 90% in the hospital, and slightly lower than 50% in the PCC and the communities. CONCLUSION: Improved procedures for PTB detection are required in the studied area to adequately control the disease and to provide therapy to affected patients. PMID- 11897175 TI - Age-related changes in risk factor effects on the incidence of coronary heart disease. AB - PURPOSE: The purpose of this report is to examine the potential for risk factor effects on the incidence of CHD to change over a broad range of ages from middle adulthood to late-life. METHODS: Findings are based on repeated risk factor measurements at four examinations over a 26-year period in men enrolled in the Honolulu Heart Program. After each examination, six years of follow-up were available to assess risk factor effects as the cohort aged from 45 to 93 years. RESULTS: Based on 18,456 person intervals of follow-up, 677 men developed CHD (3.7%). After risk factor adjustment, a positive relation between hypertension and CHD declined significantly with age (p = 0.013), primarily due to a large increase in the risk of CHD in elderly men (75 to 93) without hypertension. Effects of total cholesterol on CHD also seemed to decline with advancing age, although changes were not statistically significant. In contrast, men with diabetes had a consistent 2-fold excess risk of CHD across all age groups, while a positive association with body mass index in younger men (45 to 54) became negative in those who were the oldest (75 to 93). Due to infrequent smoking in the elderly, associations between smoking and CHD weakened with age. In the oldest men (75 to 93), alcohol intake was unrelated to CHD, while effects of sedentary life-styles on promoting CHD appeared stronger than in those who were younger. CONCLUSION: Findings suggest that changes in risk factor effects on the incidence of CHD with advancing age may require updated strategies for CHD prevention as aging occurs. PMID- 11897176 TI - A review of genetic polymorphisms and prostate cancer risk. AB - In this study, we review a variety of genetic polymorphisms that may have an etiologic role in prostate cancer. We include associations identified in molecular epidemiology studies and the consistency of findings reported to date. Suggestions for further research are also offered. For the purposes of this review, we identified relevant articles through a MEDLINE search for the period of January 1987 through March 2001. The searches were limited to articles published in English. Medical subject headings were used to scan titles, abstracts, and subject headings in the databases using the keywords "prostate neoplasms," "genetics," and "polymorphisms." PMID- 11897177 TI - Student nurses in Taiwan at high risk for needlestick injuries. AB - PURPOSE: To describe the prevalence and characteristics of needlestick injuries (NSI) in student nurses in Taiwan. METHODS: A self-administered questionnaire was completed by 931 student nurses from 16 hospitals randomly selected from the 132 accredited hospitals. RESULTS: The questionnaire was completed by 708 of 931 students who were contacted for participation in this study. NSI during internship was reported by 61.9% (438/708) of students, of whom 14.2% (62/438) made a formal report. The majority (70.1%) of NSI occurred in the patient's room. Hollow-bored needles contributed to half (219/438) of the NSIs of which 86.8% were syringe needles. Just over half (53.2%) of those items involved in NSIs had been used on patients. Of the hollow-bored needles involved in NSIs, 21.5% had been used on a patient with an infectious disease. Vaccination against hepatitis B virus (HBV) was lacking in 47.6% of students. CONCLUSIONS: NSIs and non reporting of NSIs were highly prevalent in nursing students. More intensive education programs should be directed at students to increase their awareness of and compliance with Universal Precautions (UP) before commencing their practical work experience. Students need to practice prompt post-exposure evaluation so that the need for early intervention can be assessed. In addition, any public health and infection control strategy should include a universal catch-up HBV vaccination program among students before commencement of internship. PMID- 11897178 TI - Does coffee protect against liver cirrhosis? AB - PURPOSE: To evaluate the relation between consumption of coffee and other methylxanthine-containing beverages and liver cirrhosis. METHODS: A hospital based case-control study of digestive tract and liver diseases was conducted in Greater Milan, Italy, including 101 cases with liver cirrhosis and 1538 controls. RESULTS: Compared with coffee non-drinkers, the multivariate odds ratio (OR) was 0.77 for one cup of coffee per day, 0.57 for two, and 0.29 for three or more. The OR for 40 years of coffee consumption or more was 0.45. Trends in risk were significant for both number of cups and duration of coffee drinking. No significant association was observed with decaffeinated coffee, tea and cola containing beverages. The relation between coffee consumption and liver cirrhosis was not attributable to confounding and was observed across strata of tobacco, alcohol, and other major covariates of interest. In particular, an inverse relation was observed also in subjects reporting moderate alcohol drinking. CONCLUSIONS: The present study confirms, and further quantifies, the existence of an inverse association between coffee consumption and liver cirrhosis. However, the metabolism of caffeine is impaired in fasting subjects with liver cirrhosis, and the association could be due to a reduction of coffee drinking in subjects with liver cirrhosis. PMID- 11897180 TI - Perinatal development of control systems in birds. PMID- 11897179 TI - African American smokers interested and eligible for a smoking cessation clinical trial: predictors of not returning for randomization. AB - PURPOSE: Recruitment is often the rate-limiting step in conducting clinical trials among ethnic minorities. Little is known about participants who consent and enroll into a trial, but do not return for randomization. Why participants fail to return for randomization is largely unknown. METHODS: We compared 287 enrolled African American smokers who did not return for randomization, to the 500 who returned and were randomized to participate in a clinical trial for smoking cessation in African Americans. Analyses were conducted to identify variables associated with not returning for randomization. RESULTS: Univariate comparisons found the nonrandomized group to be significantly different from those randomized. Logistic regression showed younger age, less readiness to quit, having been proactively recruited, lacking a regular source of health care, believing that they will be smoking in 6 months, less church attendance, and a lower literacy level to be jointly related with not returning for randomization. CONCLUSIONS: African American participants who did not return for randomization into a clinical trial were different from those who did. Better understanding of these factors may allow researchers to target recruitment efforts resulting in enhanced accrual in clinical trials and increased efficiency. PMID- 11897181 TI - Influence of long-term changes in incubation temperature on catecholamine levels in plasma of chicken embryos (Gallus gallus f. domestica). AB - Catecholamine concentrations were determined from day 18 to 21 of incubation (D18, D21) in developing chicken embryos. The control group was continuously incubated at 37.5 degrees C. The eggs of the two other groups were incubated at 37.5 degrees C until day 14. In the cold group, temperature was decreased to 35.0 degrees C and in the warm group, incubation temperature was increased to 38.5 degrees C for the remainder of incubation. Plasma catecholamine concentrations were measured in eggs exposed to a change in incubation temperature for 4, 5, 6 and 7 days. Embryos in the warm group had dopamine (DA) and noradrenaline (NA) concentrations that were significantly higher than in the control group. On the contrary, eggs incubated at the cooler temperature had hormone levels that were significantly lower than in the control group. Adrenaline (A) levels in the two experimental treatments were significantly lower compared to control eggs. Temperature modulated the time needed for development. Chicken embryos are supposed to hatch on day 21. However, on day 20, NA concentration in the cold incubated group was too low to fulfill its essential physiological function, whereas in the warm group, the NA concentration seems to be sufficient. Long-term exposure to altered incubation temperature affects the quantitative catecholamine concentration during development, but the relative proportion of each catecholamine remained constant. PMID- 11897182 TI - Chronic hypoxia alters the physiological and morphological trajectories of developing chicken embryos. AB - Chicken embryos were chronically exposed to hypoxia (P(O(2)) approximately 110 mmHg) during development, and assessed for detrimental metabolic and morphological effects. Eggs were incubated in one of four groups: control (i.e. 151 mmHg), or treated with continuous 110 mmHg (15% O(2)) during days 1-6 (H1-6), 6-12 (H6-12), or 12-18 (H12-18) with normoxia during the remaining incubation. Metabolism (V(O(2))), body mass, hemoglobin (Hb) and hematocrit (Hct) were measured in embryos on days 12 and 18 of incubation and in day-old hatchlings. Ability to maintain V(O(2)) was acutely measured during a step-wise decrease in P(O(2)) from normoxia to hypoxia (55 mmHg). On day 12, V(O(2)) of H1-6 eggs were significantly lower than in the control and H6-12 eggs. P(crit) in H6-12 eggs was lower than in control and H1-6 eggs. Body mass of H1-6 and H6-12 embryos on day 12 was significantly lower than in control embryos, while in H6-12 embryos, Hct and Hb were higher. On day 18, H6-12 embryos had significantly lower V(O(2)) than control eggs. Body mass of H6-12 and H12-18 embryos was significantly lower than control embryos. Hct and Hb did not differ between treatments. In hatchlings, mass, Hb and Hct had returned to values statistically identical to controls. However, H6-12 embryos had significantly lower V(O(2)). Long-term hypoxia altered V(O(2)) when hypoxic incubation occurred during the middle third of incubation, but not during earlier or later incubation. Thus, chronic hypoxic exposure during critical periods in development altered the developmental physiological trajectories and modified the phenotypes of the developing embryos. PMID- 11897183 TI - Pre- and postnatal energetics of the North Island brown kiwi (Apteryx mantelli). AB - In four eggs and four chicks of the North Island brown kiwi (Apteryx mantelli) we measured pre- and postnatal oxygen consumption rate (VO(2)) daily from day (d)-75 (prior to hatching) until d+25 (after hatching). The increase of embryonic VO(2) reaches a plateau phase between d-22 and d-5 (0.113 ml O(2)/g/h=59.6% of allometrically expected value of a typical 416-g egg). Mean total O(2) uptake per egg (43.01 l O(2)) corresponds to an energy turnover rate of 2.04 kJ/g during embryonic development. This is nearly identical to the expected value for all birds (2.00+/-0.8 kJ/g). Hence, the kiwi neither 'gained nor lost energy' (Calder, 1979.Biosci. 8, 461-467) by its extreme prolongation of incubation time; it is as efficient as other avian embryos. The kiwi embryo expends only approximately 17% (847 kJ) of the energy originally stored in the egg (4942 kJ). Forty-eight percent of the egg's initial yolk mass can be found as spare yolk in the hatchling and can serve as the chick's sole source of energy and substrate for tissue production for up to at least 17 days after hatching. PMID- 11897184 TI - Facultative and obligatory thermogenesis in young birds: a cautionary note. AB - A brief overview on thermogenic mechanisms in young precocial birds is given. While shivering thermogenesis is well documented in these birds, evidence for a facultative non-shivering component of heat production, comparable to that found in the brown adipose tissue of mammals, is ambiguous. One reason for this is the confusion between thermoregulatory and obligatory thermogenesis. In particular, the existence of a thermogenic reaction, even a futile one, does not by itself constitute proof of true thermoregulatory non-shivering thermogenesis. More probably, such a reaction is another obligatory component of heat production. Heat increment of feeding and motor activity are classical examples of such mechanisms. Thermogenesis arising from such mechanisms can often be adaptively used by the thermoregulatory systems in young birds, as well as in adults. PMID- 11897185 TI - Metabolic responses of chicken and muscovy duck embryos to high incubation temperatures. AB - Oxygen consumption, heat production (HP) and core temperature (T(af)) were measured over 3 h in 20-34-day-old Muscovy duck and 12-21-day-old chicken embryos at ambient temperature (T(a)) of 37.5 degrees C and thereafter for 3 h at T(a) of 39.0 degrees C. At 37.5 degrees C T(a), HP increased with age in avian embryos of both species, following an exponential function. In muscovy duck embryos, a plateau phase occurred between D29 and D32; in chicken embryos, a similar plateau occurred between D19 and D21. T(af) rose in accordance with HP, and the relationships between T(af) and HP could be described by significant linear regressions in both species. Mostly, HP increased in embryos of both species during heat load, but by less than calculated by the van't Hoff rule; however, there was often also a decrease in HP under these conditions. Obviously, in avian embryos high T(a) causes a down-regulation of HP mediated by active thermoregulatory mechanisms. This is in agreement with data describing the influence of hyperthermia on HP in the postnatal period of birds and mammals. Because of this, the term 'second chemical thermoregulation' defined by Gelineo [C. R. Soc. Biol. (1936) 122 337] for birds and mammals should also be used for avian embryos. PMID- 11897186 TI - Ontogeny of thermoregulation in precocial birds. AB - The aim of this paper is to summarise the results of earlier experiments on thermoregulation and heat balance in birds, to present new results concerning thermoregulation during the perinatal period in precocial embryos and to develop a model of the ontogeny of thermoregulation over the whole lifespan of birds. The ontogeny of thermoregulation in precocial birds is characterised by three phases with different efficiency of the system. In the prenatal phase, all control elements of the thermoregulatory system can function, but the efficiency of the system is low. It is postulated that endothermic reactions during the prenatal period do not have a proximate (immediate), but rather an ultimate influence on the efficiency of thermoregulation. They may support adaptivity to expected environmental conditions and may be involved in epigenetic adaptation processes. During the early postnatal phase, the thermoregulatory system develops and matures. Summit metabolism and resting metabolic rate and their thermoregulatory set points increase. Preferred temperature is significantly different during different behavioural activities. The phase of full-blown homeothermy starts at approximately the 10th day of life. It is characterised by an activation order of thermoregulatory control elements and by secondary chemical thermoregulation. The influence of thermal and non-thermal climatic factors on heat production and heat loss may be described by mathematical models. PMID- 11897187 TI - Ontogeny of thermoregulatory mechanisms in king penguin chicks (Aptenodytes patagonicus). AB - The rapid maturation of thermoregulatory mechanisms may be of critical importance for optimising chick growth and survival and parental energy investment under harsh climatic conditions. The ontogeny of thermoregulatory mechanisms was studied in growing king penguin chicks from hatching to the full emancipation observed at 1 month of age in the sub-Antarctic area (Crozet Archipelago). Newly hatched chicks showed small, but significant regulatory thermogenesis (21% rise in heat production assessed by indirect calorimetry), but rapidly became hypothermic. Within a few days, both resting (+32%) and peak (+52%) metabolic rates increased. The first week of life was characterised by a two-fold rise in thermogenic capacity in the cold, while thermal insulation was not improved. During the second and third weeks of age, thermal insulation markedly rose (two fold drop in thermal conductance) in relation to down growth, while resting heat production was slightly reduced (-13%). Shivering (assessed by electromyography) was visible right after hatching, although its efficiency was limited. Thermogenic efficiency of shivering increased five-fold with age during the first weeks of life, but there was no sign of non-shivering thermogenesis. We conclude that thermal emancipation of king penguin chicks may be primarily determined by improvement of thermal insulation after thermogenic processes have become sufficiently matured. Both insulative and metabolic adaptations are required for the rapid ontogeny of thermoregulation and thermal emancipation in growing king penguin chicks. PMID- 11897188 TI - Cardiac rhythms in prenatal and perinatal emu embryos. AB - Emu eggs weigh approximately 600 g and have an incubation duration (ID) of approximately 50 days. The egg mass is approximately 10-fold heavier than the chicken egg and the ID is approximately 2.5-fold longer. Daily changes in mean heart rate (MHR) of emu embryos were previously determined, but further measurement was needed to investigate the species-specific behavior of cardiac rhythm for comparison with other species. In the present study, we continuously measured the electrocardiogram of emu embryos while maintaining adequate gas exchange through the eggshell and determined instantaneous heart rate (IHR) during the last 2-7 days of incubation until hatching or death. The MHR over 1 min intervals was calculated from IHR data in order to present continuous developmental patterns of heart rate (HR) in a single graph and 24-h recordings of HR in a single panel, showing the HR trend over a prolonged period. However, neither circadian nor ultradian rhythms of HR were shown in these figures or by power spectrum analysis. The IHR distinctively fluctuated and the fluctuations were mainly comprised of three patterns of irregular HR accelerations in embryos that hatched. Respiratory sinus arrhythmia also occurred in perinatal embryos. During the final stages of the perinatal period, short-term, repeated, large accelerations of IHR appeared, which signaled imminent hatching and has been reported for chick embryos. IHR fluctuations in embryos that failed to hatch tended to become inactive towards death. PMID- 11897189 TI - Cardiac rhythms in developing emu hatchlings. AB - Six emu hatchlings were non-invasively measured for electrocardiogram (ECG) from their chest wall using flexible electrodes, and the instantaneous heart rate (IHR) was determined from ECG throughout the first week of post-hatching life. Although the baseline heart rate (HR) was low, approximately 100-200 beats per min (bpm), compared with chick hatchlings, the IHR fluctuated markedly. The fluctuation of IHR comprised HR variability and irregularities that were designated as types I, II and III in chick hatchlings and additional large accelerations distinctive of emu hatchlings. Type I was HR oscillation with a mean frequency of 0.37 Hz (range 0.2-0.7 Hz), i.e. respiratory sinus arrhythmia (RSA). From RSA, breathing frequency in emu hatchlings was estimated to be approximately half of that in chickens. Type II HR oscillation was also found in the emu; the frequency ranged from approximately 0.04 to 0.1 with a mean of 0.06 Hz, and the magnitude tended to be large compared with that of chickens. In addition to type III HRI, which was designated in chickens, large, irregular HR accelerations were characteristic of emu hatchlings. From IHR data, developmental patterns of mean heart rate (MHR) were constructed and plotted on a single graph to inspect the diurnal rhythm of MHR by visual inspection and power spectrum analysis. A circadian rhythm was not clear in the emu hatchlings, in contrast to chick hatchlings, which showed a dominant diurnal rhythm. PMID- 11897190 TI - Low-frequency oscillation of instantaneous heart rate in newly hatched chicks. AB - Instantaneous heart rate (IHR) of chickens began to fluctuate on days 13-14 of incubation and heart rate (HR) fluctuations became augmented towards hatching and increased further after hatching. IHR fluctuations of newly hatched chicks have been categorized into three types: type I HR variability (HRV), which is high frequency oscillation; type II HRV, which is low-frequency oscillation; and type III HR irregularities (HRI), which are irregular HR accelerations. The present experiment was carried out to investigate the origin of type II HR oscillations. Following previous evidence, we assumed that the low-frequency oscillation of HR in newly hatched chicks was related to thermoregulation and changed by environmental temperature. Eventually, type II HRV was produced or augmented by exposure of hatchlings to lowered ambient temperature and was abolished by exposure to elevated environmental temperature. The hatchlings that were exposed to large temperature decreases tended to increase their HR more than those exposed to small temperature decreases, and vice versa. The HR oscillation accompanied by an elevation of HR baseline in response to cooling may be a phenomenon related to thermoregulation in chick hatchlings. PMID- 11897191 TI - Development of heart rate responses to acoustic stimuli in Muscovy duck embryos. AB - Heart rate (HR) of Muscovy duck embryos (Cairina moschata f. domestica) was continuously recorded from the 21st day of incubation (E21) until hatching (E35). During that period, embryos were exposed to different acoustic stimuli (species specific maternal and duckling calls, music, rectangular and sine waves, white noise). Sudden HR changes occurred at the onset of acoustic stimulation (on response), as well as spontaneously. From E27 onwards, the response rate was significantly higher than the rate of spontaneous HR changes. The on-response rate increased further until E30. Most responses were elicited by maternal calls and music, but rarely by duckling calls. On-responses could be classified into: HR increase (36.4%), HR decrease (37.9%) and an increase in instantaneous HR variability (23.2%). The increase in HR variability occurred only in response to sounds, but not spontaneously. HR increases were mainly observed when the baseline HR was lower than the long-term HR trend. On-response duration was no longer than 3 min in 90% of all observations. The hourly mean HR and standard deviation did not change, even during phonoperiods composed of several sound patterns and lasting several hours. We conclude that Muscovy duck embryos are able to perceive exogenous acoustic stimuli, and that the acousto-sensory- >cardiac axis is functional from E27. PMID- 11897192 TI - Development of respiratory rhythms in perinatal chick embryos. AB - In chick embryos, gas exchange takes place via the chorioallantoic membrane (CAM) and the lungs at approximately 1 day prior to hatching. The present study was designed to elucidate the development of respiratory rhythms in the chick embryo during the whole pipping (perinatal) period with a condenser-microphone measuring system. The microphone was hermetically attached on the eggshell over the air cell on day 18 of incubation. It first detected a cardiogenic signal (i.e. acoustocardiogram), and then beak clapping and breathing signals (acoustorespirogram, ARG). The first signals of lung ventilation appeared intermittently and irregularly approximately once per 5 s among the clapping signals after the embryo penetrated its beak into the air cell (internal pipping, IP). The respiratory rhythm then developed irregularly, with a subsequent more regular rate. The envelope pattern of breathing from the onset of IP through external pipping (EP) to hatching was constructed by a specially devised procedure, which eliminated external and internal noises. The envelope patterns indicated that the IP, EP and whole perinatal periods of 10 embryos were 14.1+/ 6.4 (S.D.), 13.6+/-4.0 and 27.6+/-5.4 h, respectively. In addition, they also indicated the period of embryonic hatching activity (i.e. climax) which was 48+/ 19 min. The development of respiratory rhythm was also shown by the instantaneous respiratory rate (IRR) which was designated as an inverse value of two adjacent ARG waves. PMID- 11897193 TI - Early development of neuronal hypothalamic thermosensitivity in birds: influence of epigenetic temperature adaptation. AB - The aim of the study was to investigate the prenatal influence of different incubation temperatures on the early postnatal development of neuronal hypothalamic thermosensitivity in birds. The experiments were carried out in brain slices of 1-, 5- and 10-days-old Muscovy ducklings incubated at 35, 37.5 (control) or 38.5 degrees C during the last week of incubation. Firing rate of neuronal activity was recorded extracellularly during sinusoidal temperature changes. The results show that the temperature experienced prenatally has a clear influence on postnatal neuronal hypothalamic thermosensitivity. For instance, at the 10th day post-hatching, exposure to the cooler prenatal incubation temperature resulted in elevated neuronal hypothalamic warm sensitivity through an increased proportion of warm sensitive neurons and a reduced proportion of cold sensitive neurons in comparison with the control group. Exposure to the warmer prenatal incubation temperature induced the opposite effect. In these age group changes in neuronal hypothalamic thermosensitivity occur in relation to the prenatal temperature experienced (proximate adaptive). During the first days of life, prenatal temperature load induced a significant change in the thermosensitivity of hypothalamic neurons which was independent of the direction of change in incubation temperature in comparison with control conditions (proximate non-adaptive). Changes in the thermosensitivity of hypothalamic neurons after prenatal temperature experiences observed in all age groups may be the result of epigenetic temperature adaptation. PMID- 11897194 TI - Sex dimorphism in the avian arginine vasotocin system with special emphasis to the bed nucleus of the stria terminalis. AB - The avian neuropeptide arginine vasotocin (AVT) originally characterized as the antidiuretic hormone (, Endocrinol. 66, 860-871) is produced by neurosecretory cells within the brain. Numerous neuroanatomical studies that employed immunocytochemical and in situ hybridization techniques revealed such cells in the following anatomical brain locations: (a) preoptic area including supraoptic nucleus; (b) paraventricular nucleus; (c) the bed nucleus of the stria terminalis (BnST) (, J. Hirnforsch. 27, 559-566;, J. Neuroendcrinol. 5, 281-288;, Cell Tiss. Res. 287, 69-77;, J. Comp. Neurol. 369, 141-157). The BnST which influences reproduction and sexual behavior shows sex differences in morphology, steroid responsiveness and synthesis of neuropeptides including AVT (, Brain Res. 657, 171-184). AVT is the main endocrine regulator of fluid balance in avian species and, in addition, is involved in oviposition in these species. Our recent studies clearly demonstrated that AVT secretion after osmotic stimulation is sexually dimorphic. In order to investigate whether AVT is expressed and synthesized in the BnST in a sexually dimorphic manner we have used in situ hybridization technique and immunocytochemistry to analyze AVT gene expressing neurons in the parvocellular (small-celled nulei) BnST of adult male and female chickens. In cocks, AVT peptide-containing neurons were detected in the parvocellular BnST and the lateral septal area, whereas no AVT immunoreactive neurons were detected in the corresponding regions of the hen. Even after osmotic stimulation AVT gene expression in neurons of the parvocellular BnST of hens was not upregulated (, Cell Tiss. Res. 287, 69-77). These results demonstrate: (a) AVT gene expression in the BnST of chickens; and (b) a strong sexual dimorphism in this region. Furthermore, AVT synthesis is regulated on the transcriptional level independent from osmotic stimuli. Thus, sex steroids might be the main regulator of AVT gene expression in the BnST. In this paper we not only review the sexual dimorphic vasotocinergic system in the BnST, we also focus on the ontogeny of sex differences and the role of gonadal hormones in organization and retention of these differences. PMID- 11897195 TI - Developmental endocrinology of the reproductive axis in the chicken embryo. AB - In mammals, the phenotype of the homogametic sex develops in the (relative) absence of steroids and the phenotype of the heterogametic sex is imposed by the early action of steroids. In contrast, the heterogametic sex in avian species is the female and the presence of estrogens and their receptors plays a crucial role in female sexual differentiation. The time- and sex-dependent expression of enzymes involved in steroidogenesis which determine the ratio of androgens/estrogens produced by the gonads has been extensively investigated during the last 5-6 years. These results all show that the lack of estrogen synthesis in the male appears to be due to the extremely low levels of 17beta hydroxysteroid dehydrogenase and P450aromatase expression. In females, extensive expression of the aromatase gene (around day 5-6 of incubation), leading to estrogen synthesis, and specific expression of the estrogen receptor-mRNA in the left gonad results in the development of a functional left ovary. Other sex differences can be found in the expression of the inhibin subunit genes in gonads of chicken embryos and in circulating concentrations of inhibin, follicle stimulating hormone (FSH) and steroids. Sex reversal attempts have been made by varying incubation temperatures, by using anti-estrogens, androgens, aromatase inhibitors and synthetic steroids. In ovo administration of a sex steroid hormone or an inhibitor of endogenous sex steroid synthesis can cause phenotypical sex reversal. All these experiments show that the development of gonads in birds is very sensitive to changes in the embryonic hormonal environment, sometimes resulting in changes of postnatal reproduction and even growth. PMID- 11897196 TI - Indicators of functional differentiation of the chick embryonic kidney. AB - Relevant indicators of the functional capability of the embryonic kidney were tested in the chick mesonephros chosen as an ideal model accessible to direct observation in vivo. Evidence of glomerular filtration (GF) was checked up by the arterial injection of 2% lissamine green (LG) followed by measurement of the LG passage time on days 5, 6 and 7. Presence of the electrogenic transport was investigated by determining the transepithelial potential difference (TPD) which distinguished proximal and distal tubules of the 6-day nephrons. GF and tubular reabsorption could be demonstrated from day 5 by the storage of trypan blue (TB) in proximal tubules after intra-amniotic administration of the dye. The distribution of tubule staining corresponded to the proximal-distal gradient of the nephron differentiation. Activities of embrane enzymes, alkaline phosphatase and 5'-nucleotidase, were detected from day 4. They preceded the ultrastructural maturation in the differentiating proximal tubule epithelia. A semiquantitative evaluation of enzyme activities by the method of measuring of the minimum incubation time (MIT) together with the TB storage, appeared reliable and relevant indicators of the functional properties of mesonephric nephrons, suitable for distinguishing between more and less advanced stages of the nephron development. PMID- 11897197 TI - Rhythmic contractions in chick amnio-yolk sac and snake amnion during embryogenesis. AB - The rhythmic movements of fetal membranes in chick and reptile embryos were studied to explore the developmental role of the extra-embryonic motor activity. In the snakes Lamprophis fuliginosus and Elaphe radiata, rhythmic contractions of amnion inside the developing egg were recorded from the 11th incubation day until pre-hatching stages (ca. day 60-72). The duration of these contractions averaged 2.02+/-0.27 min. The frequency ranged from 2 to 6 per 10 min and averaged 4.61+/ 0.57 per 10 min. A tendency of frequency to increase toward the end of embryogenesis was observed. Lowering the temperature from 28 to 20 degrees C significantly decreased the frequency of amnion contractions to 2.85+/-0.91 per 10 min. The isolated snake amnion retained its capacity for spontaneous contraction. Noradrenaline inhibited, acetylcholine stimulated and serotonin did not affect the rhythmic activity of the isolated snake amnion. Similar effects were found when these agents were applied into the snake amniotic cavity. In the chick, yolk sac rhythmic contractions were recorded from the fifth until the 12th incubation days. The duration of these contractions ranged from 15 to 60 s, their frequency averaged 11.8+/-3.18 per 10 min and depended on temperature. The low temperature threshold was approximately 30 degrees C. After surgical removal of the amnion and embryo, the yolk sac continued contracting inside the egg. The yolk sac rhythmic contractions likely participate in the space movement of the embryo inside the egg during embryogenesis. PMID- 11897198 TI - Aerial and aquatic respiration of the Australian desert goby, Chlamydogobius eremius. AB - Physiological, anatomical and behavioural adaptations enable the Australian desert goby, Chlamydogobius eremius, to live in mound springs and temporary aquatic habitats surrounding the south-eastern rim of the Lake Eyre drainage basin in the harsh inland of Australia. This study describes the desert goby's respiratory and metabolic responses to hypoxic conditions and its use of buccal air bubbles for gas exchange at the water surface. Oxygen consumption for C. eremius is significantly higher in water than in air under normoxic and hypoxic conditions. In water, total oxygen consumption (V(O(2))) increases from normoxic conditions (253 microl g(-1) h(-1)) to 8% ambient O(2) concentration (377 microl g(-1) h(-1)), then decreases with increasing hypoxia of 4% O(2) (226 microl g(-1) h(-1)) and at 2% O(2) (123 microl g(-1) h(-1)). In air (fish were moist but out of water), V(O(2)) progressively decreases from normoxic conditions to hypoxic conditions (21% O(2), V(O(2)) is 169 microl g(-1) h(-1) to 39 microl g(-1) h(-1) at 2% O(2)). These data indicate oxygen-conforming patterns with increasing hypoxia both in air and in water below 8% O(2). In water, opercular movement rates remain unchanged with increasing hypoxia (139 min(-1) at 21% O(2), 154 min( 1) at 8%, 156 min(-1) at 4% and 167 min(-1) at 2%) but in air, opercular movement rates are significantly lower than in water, corresponding with the lower metabolic rate (71 min(-1) at 21% O(2), 53 min(-1) at 8%, 96 min(-1) at 4% and 64 min(-1) at 2%). Chlamydogobius eremius can use a buccal air bubble for aerial O(2) uptake, most probably in response to increased aquatic hypoxia. In air, C. eremius relies more on the buccal bubble as an oxygen source with increasing hypoxia up to an ambient O(2) of 4% (7.1% of V(O(2)) at 21% O(2); 14.5% at 8% O(2); and 27.1% at 4% O(2)), then when the available supply of O(2) is further reduced, it decreases (15% of V(O(2)) at 2% O(2)) and respiration across the skin again makes a higher relative contribution. The Australian desert goby has a higher metabolic rate in higher salinities (336 microl g(-1) h(-1) in 35 ppt, 426 microl g(-1) h(-1) in 70 ppt) than in freshwater (235 microl O(2) g(-1) h(-1)), presumably because of the increased metabolic cost of osmoregulation. There was no significant difference in V(O(2)) for fish in air that had come from varying salinities. PMID- 11897199 TI - Water content, body weight and acid mucopolysaccharides, hyaluronidase and beta glucuronidase in response to aestivation in Australian desert frogs. AB - This study investigates the effects of aestivation on body water content, body mass, acid mucopolysaccharide (AMPS) and some of its degrading enzymes in different tissues for some Australian desert frogs. The AMPS component of the liver, kidney, skin and cocoon alter during aestivation to help retain water, which is unchanged in most tissues of all frog species, and to protect the frogs from desiccation during extended periods of aestivation. Hepatic AMPS was unaltered in Cyclorana maini, C. platycephala and Neobatrachus sutor but increased significantly after 2 months of aestivation in C. australis. The level of AMPS in the kidney was elevated in all four frog species after 5 months of aestivation. Skin AMPS content in the skin of awake frogs decreases with aestivation period and increases in the cocoon. AMPS in the cocoon probably works as a cement between the cocoons' layers and its physical presence presumably contributes to preventing water flux. Changes in AMPS content in different tissues were accompanied by significant changes in both hyaluronidase and beta glucuronidase activities, which play an important role in AMPS metabolism. Alcian blue staining of control and digested skin of C. australis and C. platycephala with testicular hyaluronidase indicated the presence of AMPS, concentrated in a thin layer (called ground substance, GS) located between stratum compactum and stratum spongiosum, and acid mucin concentrated in the mucous glands and in a 'tubular' structure which could be observed in the epidermal layer. Hyaluronidase digestion of the cocoon slightly changed the Alcian Blue colour, suggesting the presence of a large amount of acid mucin similar to that found in the skin mucous gland. The results of this study present data for the redistribution of AMPS, which may help in reducing water loss across the cocoon and reabsorption of water in the kidney during aestivation. PMID- 11897200 TI - Physiological roles of free D- and L-alanine in the crayfish Procambarus clarkii with special reference to osmotic and anoxic stress responses. AB - Under hyper-salinity stress from freshwater to 17 and 25 ppt seawater, red swamp crayfish Procambarus clarkii largely accumulated D- and L-alanine together with glycine, L-glutamine, and L-proline in both muscle and hepatopancreas. The increases of D- and L-alanine in muscle were the highest in all amino acids and reached 6.8- and 5.4-fold, respectively, from freshwater to 25 ppt seawater. These results indicate that both D- and L-alanine are the most potent osmolytes for intracellular isosmotic regulation in crayfish as well as other crustaceans thus far examined. Under anoxia stress below 0.1 mg/l dissolved oxygen for 12 h and subsequent recovery in normoxia for 12 h in freshwater, 17 and 25 ppt seawater, muscle ATP decreased dramatically in all salinity levels and almost depleted in seawater. Along with the decrease of muscle glycogen level, the significant increase of L-lactate was found in muscle, hepatopancreas, and hemolymph for each salinity level, suggesting the transport of L-lactate from muscle into hepatopancreas via hemolymph. Under anoxia, D- and L-alanine also largely increased in both muscle and hepatopancreas for each salinity level. The increase was much higher in seawater than in freshwater. Thus, both D- and L alanine are possible to be anaerobic end products during prolonged anaerobiosis of this species. PMID- 11897201 TI - Fasting and lactation effect fat-soluble vitamin A and E levels in blood and their distribution in tissue of grey seals (Halichoerus grypus). AB - Grey seals among other phacoids represent a good model to study the mobilisation, transfer and deposition of fat-soluble components such as vitamins in lactating females and suckling pups because during the lactation period mothers may fast completely while secreting large quantities of high fat milks, and pups deposit large amounts of fat as blubber. The level of vitamins A and E in different tissues (liver, adipose tissue, kidney, heart, skeletal muscle, testis) and blood plasma of adult grey seal females and males changed as a result of fasting and lactation; changes were also observed in pups. The most obvious effects were a significant increase of retinol and a decrease of vitamin E levels in plasma of females with the onset of lactation as well as a substantial decrease in liver vitamin E. In suckling pups both retinol and vitamin E levels in plasma increased with the onset of suckling; after weaning no changes in retinol but a significant decrease in plasma vitamin E was observed. While liver vitamin A levels tended to be unaffected by suckling or post-weaning fast, liver vitamin E levels increased with the uptake of milk substantially (P<0.01) and returned at weaning to low levels similar to that in fetuses. Adipose tissue levels of vitamin A and E in both females and pups were only marginally affected by lactation, suckling or post-weaning fast. Results indicate that both plasma and liver levels of vitamin A and E are affected by the mobilisation, absorption and deposition of these components during lactation in seals to a much greater extent than adipose tissue, from which fat-soluble vitamins are mobilized at rates similar to that of lipids. PMID- 11897202 TI - Growth rates of Chinese and American alligators. AB - Growth rates in two closely related species, Alligator mississippiensis (American alligator) and Alligator sinensis (Chinese alligator), were compared under identical conditions for at least 1 year after hatching. When hatched, Chinese alligators were approximately 2/3 the length and approximately 1/2 the weight of American alligator hatchlings. At the end of 1 year of growth in captivity in heated chambers, the Chinese alligators were approximately 1/2 as long and weighed approximately 1/10 as much as American alligator yearlings. When the animals were maintained at 31 degrees C, Chinese alligator food consumption and length gain rates dropped to near zero during autumn and winter and body weights decreased slightly, apparently in response to the change in day length. At constant temperature (31 degrees C), food consumption by American alligators remained high throughout the year. Length gain rates in American alligators decreased slowly as size increased, but were not affected by photoperiod. Daily weight gains in American alligators increased steadily throughout the year. In autumn, provision of artificial light for 18 h a day initially stimulated both length and weight gain in Chinese alligators, but did not affect growth in American alligators. Continuation of the artificial light regimen seemed to cause deleterious effects in the Chinese alligators after several months, however, so that animals exposed to the normal light cycle caught up to and then surpassed the extra-light group in size. Even after removal of the artificial light, it was several months before these extra-light animals reverted to a normal growth pattern. These findings may be of interest to those institutions engaged in captive growth programs intended to provide animals for reintroduction to the wild or to protected habitat. PMID- 11897203 TI - The effect of vitamin C on the non-specific immune response of the juvenile soft shelled turtle (Trionyx sinensis). AB - The study was conducted to determine the effect of supplementation vitamin C in dietary non-specific immunity in juvenile soft-shelled turtles. The soft-shelled turtles were fed with vitamin C supplementation at dosages of 0, 250, 500, 2500, 5000 and 10000 mg/kg diets, respectively, for 4 weeks. The results showed that there were no differences in the phagocytosis of blood cells, serum bacteriolytic activity and bactericidal activity among soft-shelled turtles fed with vitamin C supplementation in 0-500 mg/kg diets. However, firm indicators were significantly enhanced in soft-shelled turtles fed with vitamin C supplementation at 2500 mg/kg diets compared with those fed at 0 and 250 mg/kg diets. The soft-shelled turtles fed with 5000 mg/kg diets had only significantly higher bactericidal activity than those fed vitamin C-deficient diets. The vitamin C supplementation in 10000 mg/kg diets had no notable effects on the phagocytosis, bacteriolytic activity and bactericidal activity. These results suggest that vitamin C seems have an upper and lower threshold for improving non-specific immune function, and the optimum dose was 2500 mg/kg. PMID- 11897204 TI - Plasma steroid concentrations in relation to size and age in juvenile alligators from two Florida lakes. AB - Previous studies have reported a number of physiological differences among juvenile alligators from two well-studied populations (Lake Apopka and Lake Woodruff) in north central Florida. These studies obtained alligators of similar size from each lake under the assumption that the animals were of similar age. Lake Apopka is a hypertrophic lake with a 50-year history of contamination from agricultural and municipal operations, whereas Lake Woodruff is a eutrophic lake and part of a National Wildlife Refuge that receives little point source pollution. If growth rates differ among these areas, it could be argued that differences in endocrine parameters reported previously (e.g. steroid or thyroid hormone concentrations) could be the result of differences in the animals' ages. Using growth annuli in cross-sections of femurs, we estimated the ages of juvenile alligators and compared the relationship of estradiol-17beta (E(2)) and testosterone (T) to size and age within each lake and sex. No differences were detected in the relationship between size and age between the two areas indicating similar growth rates between lakes. Plasma E(2) was positively related to size in females from Lake Apopka, and age in Woodruff females. Males from Lake Apopka had elevated plasma E(2) compared with Lake Woodruff males and did not differ from Woodruff females. No significant relationships were detected for T from either lake, and no differences in plasma T were detected among lakes or sexes. Our data indicate that both size and age can have a significant relationship with steroid concentrations. However, the relationship between steroid concentrations and size or age differed between lakes. We suggest both factors should be considered when conducting physiological studies where there is evidence to suggest growth rates may differ among populations. PMID- 11897205 TI - Underuse of aspirin in a referral population with documented coronary artery disease. AB - Despite substantial evidence that antiplatelet therapy saves lives and reduces adverse events in patients with coronary artery disease (CAD), use of the most widely available and lowest cost antiplatelet agent, aspirin, continues to be disappointingly low. In a large database of patients with known CAD, we (1) explored trends in the use of aspirin over time, (2) characterized patients most likely to take aspirin regularly, and (3) estimated the effectiveness of aspirin use by examining long-term outcomes. Using patients entered in the Duke Databank for Cardiovascular Diseases, we explored the use of aspirin from 1969 to 1999. More than 25,000 patients were sent a questionnaire that included several questions about medication use, including 1 question specifically about aspirin. Patients who failed to respond to the questionnaire received a follow-up telephone call. Aspirin use increased substantially over the most recent 4 years in the study, from 59% in 1995 to 81% in 1999. Predictors of aspirin use included younger age, male sex, being a nonsmoker, and having had a myocardial infarction or revascularization procedure. Patients who never took aspirin had a risk ratio for death of 1.85 compared with patients who regularly took aspirin. Despite the well-known beneficial effects of aspirin, too many patients without contraindications to aspirin fail to take it regularly. The health care system currently lacks effective methods to ensure that patients who have CAD have adequate follow-up concerning aspirin use. PMID- 11897206 TI - Association of fibrinogen and lipoprotein(a) as a coronary heart disease risk factor in men (The Quebec Cardiovascular Study). AB - Fibrinogen has been prospectively found to correlate with coronary heart disease (CHD) but a similar association has not been well established for lipoprotein (a) (Lp(a)). Plasma lipids, Lp(a), and fibrinogen levels were measured in 2,125 men (aged 47 to 76 years) who were free of clinical CHD. During a 5-year follow-up period, 116 first CHD events were documented. Men with CHD were older, smoked more, had a higher prevalence of diabetes, and higher levels of systolic blood pressure, cholesterol, low-density lipoprotein cholesterol, Lp(a), and fibrinogen, and lower plasma high-density lipoprotein cholesterol levels. Only fibrinogen levels in the upper tertile of the distribution compared with the lower tertiles were associated with a significant risk of CHD (adjusted risk ratio 2.5; 95% confidence interval [CI] 1.4 to 4.2; p = 0.0010). Such an association was not observed with Lp(a). To assess a possible relation between fibrinogen and Lp(a) to the risk of CHD events, men were assigned to 1 of 4 groups according to fibrinogen median levels and a Lp(a) cut-off level of 300 mg/L: group 1: fibrinogen < 4.05 g/L and Lp(a) < 300 mg/L; group 2: fibrinogen < 4.05 g/L and Lp(a) > or =300 mg/L; group 3: fibrinogen > or =4.05 g/L and Lp(a) < 300 mg/L; and group 4: fibrinogen > or =4.05 g/L and Lp(a) > or =300 mg/L. Using group 1 as a reference, a significant risk ratio was only documented in group 4 (2.5; 95% CI 1.2 to 5.1; p = 0.0132). In this population, high fibrinogen levels associated with high Lp(a) levels significantly increased the risk of CHD. PMID- 11897207 TI - Comparison of efficacy and safety of atorvastatin and simvastatin in patients with dyslipidemia with and without coronary heart disease. AB - The efficacy and safety of atorvastatin 10 mg versus simvastatin 20 mg and atorvastatin 80 mg versus simvastatin 80 mg was determined in a 6-week, prospective, randomized, open-label, blinded end-point trial of dyslipidemic patients with and without coronary heart disease. A total of 1,732 patients with hypercholesterolemia and triglycerides < or =600 mg/dl (6.8 mmol/L) were randomized to receive either atorvastatin 10 mg (n = 650), simvastatin 20 mg (n = 650), atorvastatin 80 mg (n = 216), or simvastatin 80 mg (n = 216). The primary efficacy parameter was the change in low-density lipoprotein (LDL) cholesterol from baseline to week 6. Secondary efficacy parameters included the percent change from baseline to week 6 in total cholesterol, triglyceride, high-density lipoprotein (HDL) cholesterol, very-low-density lipoprotein cholesterol, apolipoprotein B, and the percent of patients achieving their National Cholesterol Education Program (NCEP) LDL cholesterol goal at study end. Atorvastatin had significantly greater reductions from baseline in LDL cholesterol than simvastatin in both comparator groups: atorvastatin 10 mg (37.1%) versus simvastatin 20 mg (35.4%) (p = 0.0097), and atorvastatin 80 mg (53.4%) versus simvastatin 80 mg (46.7%) (p <0.0001). Atorvastatin 10 and 80 mg also provided significantly greater reductions in total cholesterol, triglycerides, very-low-density lipoprotein cholesterol, and apolipoprotein B than simvastatin 20 and 80 mg, respectively (all p <0.05). All treatment groups had a significantly decreased LDL cholesterol/HDL cholesterol ratio from baseline (all p <0.0001). In both comparator groups a higher proportion of atorvastatin treated patients reached their NCEP LDL cholesterol goal compared with simvastatin. All 4 study treatments were well tolerated. PMID- 11897208 TI - Long-term safety and efficacy of a once-daily niacin/lovastatin formulation for patients with dyslipidemia. AB - Combination therapy is increasingly recommended for patients with multiple lipid disorders, especially those at high risk for coronary events. We investigated the long-term safety and effectiveness of a new drug formulation containing once daily extended-release niacin and lovastatin. A total of 814 men and women (mean age 59 years) with dyslipidemia were enrolled in a 52-week multicenter, open label study. We used 4 escalating doses (niacin/lovastatin in milligrams): 500/10 for the first month, 1,000/20 for the second, 1,500/30 for the third, and 2,000/40 for the fourth month through week 52. Dose-dependent effects were observed for all major lipid parameters. At week 16, mean low-density lipoprotein (LDL) cholesterol and triglycerides were reduced by 47% and 41%, respectively; mean high-density lipoprotein (HDL) cholesterol was increased by 30% (all p <0.001). LDL/HDL cholesterol and total/HDL cholesterol ratios were also decreased by 58% and 48%, respectively. These effects persisted through week 52, except for the mean increase in HDL cholesterol, which had increased to 41% at 1 year. Lipoprotein (a) and C-reactive protein also decreased in a dose-related manner (by 25% and 24%, respectively, on 2,000/40 mg; p <0.01 vs baseline). Treatment was generally well tolerated. The most common adverse event was flushing, which caused 10% of patients to withdraw. Other adverse events included gastrointestinal upset, pruritus, rash, and headache. Drug-induced myopathy did not occur in any patient. The incidence of elevated liver enzymes to >3 times the upper limit of normal was 0.5%. Once-daily niacin/lovastatin exhibits substantial effects on multiple lipid risk factors and represents a significant new treatment option in the management of dyslipidemia. PMID- 11897209 TI - Differentiation between LQT1 and LQT2 patients and unaffected subjects using 24 hour electrocardiographic recordings. AB - This study assesses the use of 24-hour ambulatory electrocardiographic recordings in distinguishing patients with long-QT1 syndrome (LQT1) from those with LQT2, and for distinguishing affected from unaffected patients. The diagnoses of the congenital LQT syndrome and its most common types LQT1 and LQT2 are made difficult because of the limitations of the electrocardiogram as a diagnostic tool. With an automated computerized program, Holter recordings from 15 LQT1 and 15 LQT2 patients and 43 healthy subjects (training set) were reviewed to select the best criteria using QT duration and rate dependence as well as the difference between QT end and QT apex to separate the 3 groups. Fixed criteria were then applied in blinded fashion to separate a different group of 32 genotyped patients and 16 unaffected subjects (test set). In the training set, the RR interval (100 ms), a slope value for median QT/RR curves of -0.016 separated 25 of 30 (83%) and a minimal QT end - QT apex value of 80 ms, separated 26 of 30 (87%) LQT1 patients from LQT2 patients. When all selected criteria were applied to differentiate LQT1 from LQT2 versus unaffected genotypes in the test set, 38 of 48 cases (79%) were correctly identified, whereas using the electrocardiogram alone, 60% of patients were correctly classified into 3 genotypes (p = 0.03). Combining measures for QT duration, rate dependence, and QT end - QT apex interval, derived from Holter recordings, complements the clinical differentiation between LQT1 versus LQT2 patients and between affected and unaffected persons for genotype screening purposes. PMID- 11897210 TI - Effects of eprosartan versus hydrochlorothiazide on markers of vascular oxidation and inflammation and blood pressure (renin-angiotensin system antagonists, oxidation, and inflammation). AB - Antagonists of the renin-angiotensin system, such as angiotensin type 1 (AT(1)) receptor inhibitors and angiotensin-converting enzyme inhibitors, are becoming increasingly popular agents in treating patients with systemic hypertension and minimizing organ damage. In the present study, we compared the effects of eprosartan, an AT(1) receptor inhibitor, with the diuretic hydrochlorothiazide in a group of newly diagnosed hypertensive patients with multiple risk factors for atherosclerosis. The subjects were monitored and tested at 0 and 4 weeks to determine their individual effects on vascular and inflammatory markers. Although blood pressure reduction was comparable between the 2 agents, there were notable differences in their effects on markers of inflammation and oxidation. We observed a 28% reduction in neutrophil superoxide anion generating capacity, a 34% reduction in soluble monocyte chemotactic protein-1, and a 35% reduction in soluble vascular cell adhesion molecule with eprosartan therapy (all p <0.05 from the start of therapy). In addition, eprosartan showed further benefit in its ability to increase low-density lipoprotein oxidation lag time, suggesting an increased resistance to oxidation and/or modification of low-density lipoprotein. Although hydrochlorothiazide was effective in blood pressure reduction, there were no significant changes in any of the above parameters after 4 weeks of treatment. These findings suggest that eprosartan, an AT(1) receptor inhibitor, effectively reduces systemic blood pressure and, compared with hydrochlorothiazide, suggests additional benefits in the vasculature by inhibiting mechanisms of inflammation and oxidation. PMID- 11897211 TI - Risk stratification using a combination of cardiac troponin T and brain natriuretic peptide in patients hospitalized for worsening chronic heart failure. AB - We prospectively evaluated whether the combination of admission measurements of a marker for myocardial cell injury and a marker for left ventricular overload would effectively risk stratify patients with acutely decompensated heart failure. We measured serum concentrations of cardiac troponin T (cTnT) using a second-generation assay, as well as serum cardiac troponin I (cTnI) and plasma atrial and brain natriuretic peptide (BNP) concentrations on admission in 98 consecutive patients hospitalized for worsening chronic heart failure (mean age 69 years; 5 patients were in New York Heart Association functional class II, 35 were in class III, and 58 patients were in class IV). During a mean follow-up period of 451 days, there were 37 cardiac events, including 21 cardiac deaths (14 in-hospital deaths) and 16 readmissions for worsening heart failure. In a stepwise Cox regression analysis, including these biochemical markers, age, sex, functional class, and left ventricular ejection fraction, cTnT, and BNP were found to be significantly independent predictors of both cardiac death (p <0.05) and cardiac events (p <0.01). A cTnT >0.033 microg/L and/or a BNP >440 pg/ml on admission was correlated with an incremental increase in in-hospital cardiac mortality, overall cardiac mortality, and cardiac event rate. Kaplan-Meier analysis revealed that this combination could reliably stratify the patients into low-, intermediate-, and high-risk groups for cardiac events. Measuring the combination of admission concentrations of cTnT and BNP may be a highly effective means of risk stratification of patients hospitalized for worsening chronic heart failure. PMID- 11897212 TI - Differentiation of the metabolic and vascular effects of insulin in insulin resistance in patients with chronic heart failure. AB - Chronic heart failure (HF) is associated with insulin resistance. Putative mechanisms of insulin resistance are abnormal skeletal muscle blood flow and antagonism of insulin action due to sympathetic nervous system activation. We measured insulin sensitivity, the vasoactive properties of insulin, and the association between insulin resistance and markers of neurohormonal activation in 10 patients with chronic HF and in 9 healthy controls. Noninvasive hemodynamic measurements and an hyperinsulinemic, euglycemic clamp were used. Patients were insulin resistant compared with the controls (p <0.05 for area under insulin dose response curve). Insulin infusion led to a selective increase in forearm blood flow accompanied by a decrease in mean arterial pressure and superior mesenteric blood flow. Heart rate decreased in patients but not in controls; however, when baseline measurements were controlled for, there was no difference in the overall hemodynamic response to insulin infusion between the study groups. In univariate analysis, age, serum creatinine, fasting insulin, and triglyceride levels correlated inversely with insulin sensitivity (p <0.05 for all). Cardiac output had a significant correlation with insulin sensitivity (p <0.05). On stepwise multiple linear regression analysis, only age and fasting plasma insulin emerged as significant predictors of insulin sensitivity (R(2) 0.613, p = 0.001). In particular, we found no evidence of a relation between insulin sensitivity and plasma noradrenaline. Patients with chronic HF exhibit significant metabolic insulin resistance. Insulin resistance is not secondary to failure of insulin mediated vasodilatation or sympathetic nervous system activation and is likely due to abnormalities at the level of the skeletal myocyte. PMID- 11897213 TI - Peak early diastolic velocity rather than pressure half-time is the best index of mechanical prosthetic mitral valve function. AB - Reliable screening of mechanical prosthetic mitral valve (PMV) dysfunction by transthoracic echocardiography (TTE) is mandatory because transesophageal echocardiography (TEE) cannot be routinely used. However, acoustic shadowing seriously hampers detection of PMV dysfunction with TTE, particularly regurgitation. To identify TTE indexes that can detect PMV dysfunction (regurgitation or obstruction), 134 patients (age 60 +/- 12 years, 64 men) with PMV who underwent TTE and TEE within 3 +/- 5 days were assessed. There were 73 normal and 61 dysfunctional valves (40 regurgitant, 21 obstructive). By multivariate analysis, peak E velocity was the best predictor of a dysfunctional valve. Both peak E velocity (E > or =1.9 m/s; sensitivity 92%, specificity 78%) and the ratio of velocity-time integrals of flow through the prosthesis to that of the left ventricular outflow (VTI(pmv/)VTI(lvo) > or =2.2; sensitivity 91%, specificity 74%) were successful in detecting PMV dysfunction. Although pressure half-time (PHT) readily identified PMV obstruction, it did not detect regurgitation. Logistic models including peak E velocity and VTI(pmv)/VTI(lvo) or PHT were equally successful in detecting PMV dysfunction. However, all 3 variables were needed to best distinguish among normal, obstructed, and regurgitant valves. A peak E velocity > or =1.9 m/s and VTI(pmv)/VTI(lvo) ratio > or =2.2 predicted valve regurgitation in 83% of valves when PHT was < 130 ms, and valve stenosis in 95% when PHT was >130 ms. Importantly, a peak E velocity < 1.9 m/s, VTI(pmv)/VTI(lvo) ratio < 2.2, and a PHT < 130 ms had a predictive accuracy for a normal valve of 98%. Thus, TTE Doppler indexes can be used as screening parameters of PMV dysfunction and help select patients for further diagnostic evaluation with TEE. PMID- 11897215 TI - Coronary artery disease in patients with heart failure and preserved systolic function. PMID- 11897214 TI - Diagnostic accuracy and cost-effectiveness of contrast echocardiography on evaluation of cardiac function in technically very difficult patients in the intensive care unit. AB - Echocardiographic assessment of cardiac function can be quite difficult in the intensive care unit and may require transesophageal echocardiography (TEE). We therefore compared harmonic imaging alone or in combination with contrast to TEE in 32 consecutive patients in the intensive care units who were considered technically very difficult (> or =50% of the 16 segments not visualized from any view). Excellent or adequate endocardial visualization was achieved in 13% of segments with fundamental imaging, 34% with harmonic imaging, and 87% with contrast (p < 0.0001); the latter success rate was similar to TEE (87% vs 90%; p = NS). When TEE was used as the standard, agreement in exact interpretation of wall motion increased from 48% for fundamental imaging to 58% with harmonic imaging, and reached 70% with contrast (p <0.0001). Contrast had the best sensitivity (89%) for detecting wall motion abnormalities. Estimation of ejection fraction was possible in 31% with fundamental imaging, 50% with harmonic imaging, and in 97% with contrast. Ejection fraction quantitated by contrast enhancement correlated best with TEE (r = 0.91). Cost-effectiveness analysis revealed that contrast echo was cost-effective compared with TEE in determining regional and global ventricular function, with a cost saving of 3% and 17%, respectively. Thus, contrast echocardiography provides an accurate, safe, and cost-effective alternative to TEE for evaluating ventricular function in technically very difficult studies. PMID- 11897216 TI - Could vaccinations prevent myocardial infarction? PMID- 11897217 TI - Charles Richard Conti, MD: a conversation with the editor. PMID- 11897218 TI - Comparison of long-term survival following non-Q-wave creatine kinase elevation after percutaneous coronary intervention in patients discharged on a beta blocker versus those not so treated. PMID- 11897219 TI - Systemic T-cell activation in stable angina pectoris. PMID- 11897220 TI - Volumetric intravascular ultrasound quantification of the amount of atherosclerosis and calcium in nonstenotic arterial segments. PMID- 11897221 TI - Proteinuria, serum creatinine, and outcome of percutaneous coronary intervention in patients with diabetes mellitus. PMID- 11897223 TI - Effect of atropine on heart rate turbulence. PMID- 11897224 TI - Bactericidal efficacy of sterilizing protocol for reused cardiac electrophysiology catheters. PMID- 11897222 TI - Implications of 2001 cholesterol treatment guidelines based on a retrospective analysis of a high-risk patient cohort. PMID- 11897226 TI - Causes and effects of poststenotic dilation of the pulmonary trunk. PMID- 11897225 TI - Usefulness of the supine-rest maneuver before neurohormonal sampling. PMID- 11897227 TI - Successful "pre-closure" of 7Fr and 8Fr femoral arteriotomies with a 6Fr suture based device (the Multicenter Interventional Closer Registry). PMID- 11897228 TI - Serum cardiac troponin I in acute rheumatic fever. PMID- 11897230 TI - Links between psychological sense of control and disturbed eating behavior in women with diabetes mellitus. Implications for predictors of metabolic control. AB - PURPOSE: Eating disturbances and aspects of psychological control are both repeatedly cited as significant correlates of metabolic control in diabetes mellitus (DM), yet such findings are typically discussed in separate literatures and have been criticized for overreliance on outdated constructs of psychological control when more complex means of analysis are available. METHODS: Utilizing a multidimensional control inventory, this study investigates the relationship between eating disturbance and psychological sense of control, and assesses the utility of these two constructs in predicting metabolic control in 96 women recruited from a specialist diabetes clinic. RESULTS: Despite significantly overlapping relationships between these two predictor variables and metabolic control, it is control specific to the domain of interpersonal relationships, along with eating disturbance in the form of bulimia/food preoccupation, that independently predicts level of metabolic control. CONCLUSIONS: These findings have implications for the current form and content of psychological interventions in the management of DM. PMID- 11897231 TI - Dieting frequency among college females: association with disordered eating, body image, and related psychological problems. AB - OBJECTIVE: To examine associations between dieting frequency and eating disorder behaviors, body satisfaction, and related factors. METHOD: Females (N=345) whose average age and body mass index (BMI) were 20.58 and 21.79, respectively, were grouped into three categories of lifetime dieting frequency (never, 1-5 times, or 6 or more times) and matched on current BMI across categories. RESULT: Positive associations were found between dieting frequency and eating disorder symptoms and related problems such as body dissatisfaction, current body size perception, depression, exercise preoccupation, and feelings of ineffectiveness and insecurity. Dieting frequency was inversely associated with self-esteem, ideal body size, emotional regulation, and impulse control. DISCUSSION: Independent of current BMI, frequency of dieting behaviors is strongly associated with negative emotions and problematic behaviors. As this study is correlational in nature, future longitudinal studies should ascertain the sequence of onset of these experiences. PMID- 11897232 TI - Effects of prolonged and repeated body image exposure in binge-eating disorder. AB - OBJECTIVE: The purpose of the present study was to investigate psychological mechanisms associated with prolonged and repeated body image exposure. METHOD: In an experimental design, 30 female volunteers diagnosed with binge-eating disorder (BED) (DSM-IV) and 30 non-eating-disordered controls (NC) were exposed to their physical appearance in a mirror. The confrontation procedure was guided by a standardized interview manual and took place on two separate days. Self-reported mood, appearance self-esteem, and frequency of negative cognitions were assessed repeatedly throughout the experiment. RESULTS: During body image exposure sessions, binge-eating-disordered individuals showed significantly lower mood than controls while appearance self-esteem was diminished in both groups. During the second body image exposure session, higher levels of mood and appearance self esteem were observed in both groups, and negative cognitions occurred less frequently. CONCLUSION: Results are discussed with regard to the therapeutic use of body image exposure. PMID- 11897233 TI - The role of puberty, media and popularity with peers on strategies to increase weight, decrease weight and increase muscle tone among adolescent boys and girls. AB - OBJECTIVE: The present study was concerned with the impact of pubertal development, relationships with peers and perceived pressure from the media on body dissatisfaction and body change behaviors among adolescent boys and girls. In particular, the study investigated the underresearched area of strategies to increase weight and muscle. The exploration of body change strategies among adolescent boys has been a neglected area of research. METHODS: Respondents were 1185 adolescents (527 males, 598 females) who were enrolled in Grades 7 and 9. Participants completed measures of pubertal development, media and peer influence, body dissatisfaction and strategies to lose weight, increase weight and to increase muscle. RESULTS: The findings demonstrated that girls were more likely than boys to adopt strategies to lose weight, whereas boys were more likely to adopt strategies to increase muscle tone (but not weight). For boys in both Years 7 and 9, the main predictors of body change strategies were puberty and, to a lesser extent, perceived popularity with peers. The major influences for Years 7 and 9 girls were puberty and the media, but these mainly focused on weight loss. For Year 9 girls, perceived popularity with opposite-sex peers also predicted body dissatisfaction and strategies to increase muscle tone. CONCLUSION: The implications of these findings for understanding factors related to a range of body change strategies for adolescent boys and girls are discussed. PMID- 11897234 TI - A review of psychosocial outcomes of surgery for morbid obesity. AB - There is consistent evidence to support the notion that morbid obesity poses serious risks to physical health and has a substantial impact on psychosocial well-being. Researchers agree that bariatric surgery is currently the most viable option for successful weight loss and maintenance in the morbidly obese individual. The drastic, major weight loss and alleviation of medical risks that patients typically experience post-surgically are accompanied by psychosocial changes that appear to be equally remarkable. These psychosocial changes have yet to be studied as systematically or diligently as the physical changes and therefore remain to be fully understood. This paper (1) reviews the literature of psychosocial outcomes of obesity surgery for the past 36 years; (2) provides a critical assessment of the methodology utilized; and (3) suggests future research directions. PMID- 11897235 TI - Validation of the Spanish version of the Perceived Stress Questionnaire. AB - OBJECTIVE: To validate in Spanish the Perceived Stress Questionnaire (PSQ), a questionnaire to assess stress for research purposes in psychosomatic patients. METHOD: The test was administered to a healthy population (N=174) of nursing students and health workers and to a clinical sample (N=80) of patients attending a psychiatric outpatient consultation. RESULTS: Concurrent validity: General and Recent PSQ scores correlated high with trait anxiety (r=.65), moderate with depression (r=.46) and psychological disturbance (r=.51) and poor with state anxiety (r=.22). Predictive validity: PSQ scores were higher in "psychiatric cases" than in "psychiatric noncases" (P<.01), and correlated highly with somatic symptoms of psychological origin (r=.62) in the clinical subsample. Internal consistency was 0.9 for the General and 0.87 for the Recent PSQ. Test-retest reliability of the General PSQ was 0.80. DISCUSSION: The Spanish version of PSQ presents good psychometric properties and it seems to be a valuable instrument for psychosomatic researchers. PMID- 11897236 TI - Maternal correlates of health status in adolescents with inflammatory bowel disease. AB - OBJECTIVE: People with inflammatory bowel disease (IBD) manifest great variability in health status, but little research has examined correlates of this variability, especially among adolescents with IBD. This study examined family dysfunction, maternal physical symptoms and maternal positive affect (PA) as correlates of variability in depression, pain/fatigue, functional disability and bowel movement frequency among adolescents with IBD. We also tested whether these relationships were independent of maternal negative affect (NA). METHODS: Participants were 62 adolescents with IBD (36 with Crohn's disease (CD) and 26 with ulcerative colitis [UC]) and their mothers. Mothers completed measures about the family and themselves, and adolescents provided health status measures. RESULTS: Controlling for the duration since diagnosis, we found that family dysfunction correlated significantly and positively with bowel movement frequency of the adolescent (partial r=.27), and mother's PA correlated inversely with adolescent's depression (partial r=-.30) and functional disability (partial r= .28). These relationships remained significant after controlling for maternal NA. Maternal symptoms were not related to the adolescent's health status. Analyses indicated that relationships did not differ for the two IBD subtypes. DISCUSSION: Family dysfunction and maternal PA appear to account for some of previously unexplained variance in the health status of adolescents with IBD. Family interventions should be explored to determine whether these familial and maternal factors influence the health of the adolescent, and whether improving family functioning can lead to health improvements in the children. PMID- 11897238 TI - Varicella zoster virus vasculopathy and disseminated encephalomyelitis. PMID- 11897239 TI - Cognitive dysfunction in patients with mildly disabling relapsing-remitting multiple sclerosis: an exploratory study with diffusion tensor MR imaging. AB - Previous studies assessing the magnetic resonance imaging (MRI) correlates of cognitive dysfunction in multiple sclerosis (MS) achieved conflicting results. Diffusion tensor (DT)-MRI provides metrics that are sensitive to the macro- and microscopic MS lesion load with increased specificity to the more destructive aspects of MS pathology than conventional imaging. We performed an exploratory study to assess the magnitude of the correlation between quantities derived from DT-MRI and measures of cognitive impairment in patients with relapsing-remitting (RR) MS.T2, T1, DT-MRI scans of the brain and an extensive battery of neuropsychological tests (exploring language, complex reasoning, attention and memory) were obtained from 34 RRMS patients. We measured T2 and T1 lesion volumes (LV) and brain volume. Average lesion mean diffusivity (D) and fractional anisotropy (FA) were calculated. D and FA histograms from the brain tissue (BT), the normal-appearing brain tissue (NABT), the normal-appearing white matter (NAWM) and the normal-appearing gray matter (NAGM) were also obtained. Nine patients (26.5%) were found to be cognitively impaired. Moderate correlations were found between symbol digit modalities test, verbal fluency test and 10/36 spatial recall test scores and T2 LV, T1 LV and average lesion, WBT, NABT, NAWM and NAGM values (r values ranging from -0.30 to -0.53). No correlations were found between any of the neuropsychological test scores and brain volume, average lesion FA and WBT FA.DT-MRI provides quantitative metrics that seem to reflect the severity of language, attention and memory deficits in patients with RRMS. This study also suggests that the extent and the intrinsic nature of the macroscopic lesions as well as the damage of the NAWM and NAGM all contribute to the neuropsychological deficits of RRMS patients. PMID- 11897240 TI - Encephalitis related to primary varicella-zoster virus infection in immunocompetent children. AB - INTRODUCTION: Encephalitis is a rare complication of primary varicella-zoster virus (VZV) infection in immunocompetent children. METHODS: The clinical and laboratory findings of two girls with VZV-related encephalitis are reported. RESULTS: Both children presented with focal epileptic seizures, corresponding to cortical/subcortical as well as white matter lesions. The first showed a typical vesicular skin rash. She was easily diagnosed and made a rapid recovery during acyclovir and steroid treatment. In the second girl, a preceding measles-mumps rubella virus vaccination and the absence of skin vesicles were misleading with respect to the diagnosis, which was finally proven by IgG seroconversion and intrathecal synthesis of IgG antibodies to VZV. She developed left parieto occipital tissue necrosis and recovered only transiently during initial acyclovir/steroid treatment. Eight weeks after onset, progressive white matter demyelination and the occurrence of erythema nodosum in the lower limbs necessitated a second 4-month course of oral steroids. The VZV PCR from cerebrospinal fluid was negative in both children. CONCLUSIONS: Primary VZV infection may cause severe encephalitis that may occur without skin vesicles and lead to a chronic course with systemic vasculitis. The coincidence of vaccination and neurologic diseases offers no proof per se of a causal relationship. PMID- 11897241 TI - Family history of stroke in stroke types and subtypes. AB - Many studies have provided data showing that family history of stroke (FHS) is associated with an increased risk of stroke. The association of the FHS with the various stroke subtypes has not been adequately studied. The purpose of this study was to assess the association of the FHS with the two major stroke types (cerebral haematomas and ischaemic strokes) and the four stroke subtypes (cardioembolic, large artery disease, small artery disease, and undetermined) in a Greek population. The FHS was obtained from 421 consecutive acute stroke patients and from 239 matched control subjects. Positive FHS was observed in 49% of all stroke patients compared with 28% of the control subjects [adjusted OR=2.06 (95% confidence intervals (CI) 1.42-3.00)]. Haematomas, ischaemic strokes, and from the ischaemic strokes, both large and small artery disease strokes were strongly associated with positive FHS compared with the control subjects [adjusted OR=2.06 (95% CI 9-3.04), 2.07 (95% CI 1.09-3.91), 2.05 (95% CI 1.24-3.38), and 2.76 (95% CI 1.55-4.91), respectively]. There was no difference between maternal and paternal heritable contribution.In conclusion, FHS was found in this study to be an independent risk factor for all strokes combined, for each stroke type, and for the large and small-artery disease stroke subtypes, but not for the cardioembolic and undetermined stroke subtypes. PMID- 11897242 TI - Progressive cerebral atrophy in multiple system atrophy. AB - Nine patients with multiple system atrophy (MSA) were studied based on MRI findings of cerebral hemispheric involvement. The age at onset was 56.4+/-8.6 (mean+/-S.D.) years, duration of illness at the first MRI study 2.1+/-1.1 years, duration of illness at the last study 9.7+/-2.6 years, and the follow-up duration 7.6+/-2.3 years. Controls were 85 neurologically intact persons (60.2+/-11.1 years age). In the MRI study, measurements of the ratio of each area to the intracranial area were performed for the cerebral hemisphere, frontal, temporal and parietal-occipital lobes. A significant progression of atrophy to under the normal limit was observed in the cerebrum, frontal and temporal lobes. Besides the typical pathological lesions in MSA, five autopsied patients revealed frontal lobe atrophy with mild gliosis, mild demyelination and glial cytoplasmic inclusions (GCIs). One of these patients showed remarkable frontal lobe atrophy with degenerative changes in the cerebral cortex. We observed the involvement of the cerebral hemisphere, especially the frontal lobe. PMID- 11897243 TI - Compound heterozygosity with two novel mutations in the HEXB gene produces adult Sandhoff disease presenting as a motor neuron disease phenotype. AB - Little information is available on molecular defects involved in adult Sandhoff disease presenting as motor neuron disease phenotype. We studied enzyme activities of beta-hexosaminidase (Hex) and the HEXB gene encoding the beta subunit of Hex in a family of the Japanese case. Enzyme assay with 4 methylumbelliferyl-2-acetamido-2-deoxy-beta-D-glucopyranoside revealed a reduction in Hex A and B activity in proband's leukocytes. Although the activity of both in the mother were intermediate between those of controls and the proband, only Hex B reduction determined with heat inactivation was found in the father. Analysis of HEXB gene demonstrated two novel point mutations. The first mutation, IVS2-1G>A, was located at the 3'-splice acceptor site of intron 2 derived from the mother, causing exon 3 skipping. The resultant mRNA encoded a shorter beta-chain, which may not form an active enzyme. The second mutation was a G-to-A transition in exon 13 (c.1598G>A) derived from the father and resulted in arginine-to-histidine substitution at amino acid position 533 (R533H). Expression of R533H mutation in COS-1 cells demonstrated a lack of normal Hex activity, indicating that this mutation is pathological. Compound heterozygosity of these two mutations may trigger the development of adult Sandhoff disease with a motor neuron disease phenotype. PMID- 11897244 TI - Dynamics of cerebral blood flow autoregulation in hypertensive patients. AB - In hypertensive patients, the upper and lower limits of cerebral autoregulation are shifted to higher levels. However, the dynamics of cerebral autoregulation in hypertensive patients are less well known. We compared the dynamics of cerebral autoregulation in 21 treated hypertensive patients (13 men and 8 women; mean age: 48.9+/-13.6 years) and in 21 normotensive subjects (13 men and 8 women; mean age: 51+/-14.5 years) by transcranial Doppler (TCD) of the middle cerebral artery (MCA) during the acute decrease in blood pressure induced by standing up after 2 min in squatting position. MCA maximal outline blood flow velocity (FV), blood pressure (Finapres) and end-tidal PCO2 were continuously monitored and computerised. A cerebral vascular resistance index (CR) was calculated as follows: mean arterial BP/MCA mean FV with normalised changes in CR per second during the blood pressure decrease (CR slope). The CR slope reflecting the rate of cerebral autoregulation did not differ between the two groups and within the hypertensive patients [well controlled (8 patients) and not controlled (13 patients)]. The time to maximum decrease of CR (T1) and the time to full recovery of CR after the initial drop (T2) were also similar in the two groups (controls T1: 11.3+/-3.1 s, T2: 12+/-5.9 s; hypertensive T1: 11.7+/-2.5 s, T2: 10.7+/-4.5 s) and within hypertensive patients. These findings suggest that the dynamics of cerebral autoregulation are well preserved in hypertensive patients, with no difference according to the efficiency of treatment of hypertension. PMID- 11897245 TI - Fine specificities of anti-LM1 IgG antibodies in Guillain-Barre syndrome. AB - We investigated the prevalence of anti-LM1 IgG antibody and its fine specificity in Guillain-Barre syndrome (GBS). Anti-LM1 IgG and IgM antibodies from sera of 47 patients with GBS--19 with acute inflammatory demyelinating polyneuropathy (AIDP), 27 with acute motor axonal neuropathy (AMAN), and 1 with acute motor sensory axonal neuropathy (AMSAN)--were tested. Anti-LM1 IgG antibody was detected in only one patient with AIDP, whereas it was present in seven with AMAN and in one with AMSAN. Sera from the eight IgG anti-LM1-positive patients with AMAN/AMSAN also had IgG activity against the gangliosides GM1, GM1b, GD1a, GalNAc GD1a, GD1b, or GQ1b. Anti-LM1 IgG antibodies from the AMAN/AMSAN patients cross reacted with other gangliosides, whereas IgG antibody from the AIDP patient was monospecific against LM1. Anti-LM1 IgG antibody therefore, cannot be a marker of AIDP. In addition, whether monospecific anti-LM1 IgG antibody is associated with AIDP remains to be concluded. Larger studies are needed to verify whether monospecific anti-LM1 IgG antibody could be a marker of AIDP. PMID- 11897246 TI - Double-blind crossover study of branched-chain amino acid therapy in patients with spinocerebellar degeneration. AB - To determine whether treatment with branched-chain amino acids (BCAA) can improve the condition of patients with ataxia, a double-blind crossover study of BCAA therapy was performed in 16 patients with spinocerebellar degeneration (SCD). The patients were treated with BCAA in oral doses of 1.5, 3.0, or 6.0 g or with placebo daily for 4 weeks in each study phase. The order of treatment phases (placebo or BCAA) was assigned randomly. An International Cooperative Ataxia Rating Scale (ICARS) was used to quantify the severity of symptoms of SCD. The mean ICARS score improved significantly with BCAA treatment compared with the mean pretreatment score (p<0.01). In addition, the improvement in the mean global ICARS score was significant in the middle-dose group compared with that in the placebo group (p<0.02). The estimated improvement in kinetic functions compared with pretreatment (p<0.01) was significant after treatment with BCAA, 1.5 and 3.0 g. All of the responders manifested predominantly cerebellar symptoms, especially those with spinocerebellar ataxia type 6 (SCA6). Thus, treatment with BCAA may be effective in patients with the cerebellar form of SCD. PMID- 11897247 TI - Progression and staging of Lewy pathology in brains from patients with dementia with Lewy bodies. AB - Using alpha-synuclein-immunohistochemistry, 27 brains of dementia with Lewy bodies (DLB) were investigated to identify the progression of Lewy pathology including Lewy bodies (LB) and LB-related neurites in the cerebrum. The numbers of alpha-synuclein-positive LB and LB-related neurites were semiquantitatively evaluated in the amygdala, hippocampus, entorhinal cortex, transentorhinal cortex, insular cortex, middle temporal cortex and superior frontal cortex. The results indicated that Lewy pathology within the neuron progresses first in the axonal terminal, subsequently in the cell body and finally in the dendrite, that Lewy pathology in the cerebral cortex progresses first in layers V-VI, subsequently in layer III and finally in layer II, and that Lewy pathology in the cerebrum progresses first in the amygdala, subsequently in the limbic cortex and finally in the neocortex. In addition, Lewy pathology was graded from stage I to stage IV based on the progression of Lewy pathology. The 27 brains examined were classified into 3 brains showing stage I, 11 showing stage II, 7 showing stage III and 6 showing stage IV. Comparing these stages with the pathological subtypes of DLB brains, brains of the subtype showing severe Alzheimer pathology corresponded to brains showing an advanced stage, suggesting that Alzheimer pathology exacerbates Lewy pathology. PMID- 11897248 TI - Frequency and patterns of subclinical cognitive impairment in patients with ANCA associated small vessel vasculitides. AB - We investigated the prevalence of disease-related cognitive impairment in patients with anti-neutrophil cytoplasmic antibodies (ANCA)-associated small vessel vasculitides (SVV). We studied 43 patients with ANCA-associated SVV (Wegener's granulomatosis (WG), Churg-Strauss syndrome (CSS) and microscopic polyangiitis (MP)), with no evidence of focal neurological deficits and dementia and in whom other potential causes of cognitive decline were carefully excluded. All patients underwent a detailed neuropsychological evaluation and their performances were compared with those of matched healthy controls. Patients were considered to be affected by subclinical cognitive impairment when they had abnormal results in at least two neuropsychological tests. Magnetic resonance imaging (MRI) scans of the brain were also obtained in 11 patients.The average neuropsychological test scores were not significantly different between the SVV patients and the control subjects. Thirteen patients had abnormal results in two tests (seven patients) or three or more tests (six patients). Most frequently, abnormal tests were the Rey Figure Recall (six cases), the Wisconsin Card Sorting Test (six cases), and the reaction times (eight cases). The frequency and extent of brain MRI abnormalities were higher in impaired than in unimpaired patients. This study demonstrates that 30% of clinically nondemented SVV patients can have a subclinical neuropsychological impairment, characterized by mild abstract reasoning loss, mental speed reduction and nonverbal memory impairment. MRI findings in impaired patients are consistent with the presence of an SVV-mediated subcortical damage of the brain. PMID- 11897249 TI - Deep brain stimulation of the subthalamic nucleus in PD: an analysis of the exclusion causes. AB - OBJECTIVE: Deep Brain Stimulation (DBS) of the Subthalamic Nucleus (STN) represents a proper choice for the treatment of advanced Parkinson's disease (PD). A correct selection of the patients as candidates for the surgery is essential for a good outcome. In this study, we analyzed the exclusion causes of a series of PD patients hospitalized for the selection protocol. METHODS: Ninety eight PD patients as potential candidates for the STN DBS were studied. All patients were hospitalized and underwent a clinical evaluation of the PD stage, a levodopa challenge, a MR of the brain and a neuropsychological assessment. RESULTS: The percentage of subjects considered not suitable for the surgery was 29.6%. A single cause of exclusion was present in 65.5% of not suitable patients, while multiple causes were present in 34.5%. The most frequent cause of exclusion was the finding of neuropsychological or psychic disorders (48.3%); in 37.9% of the patients, the motor disability was not severe enough to justify the surgery, while in 31%, we found relevant abnormalities at the brain MR. Three patients (10.3%) were poorly motivated for the surgery, while in three others (10.3%), we found a significant illness other than PD. CONCLUSIONS: The finding that about 30% of the PD patients potentially suitable for STN DBS presents some exclusion causes underlines the importance of a careful selection of the candidates for this surgery. PMID- 11897250 TI - White matter changes mimicking a leukodystrophy in a patient with Mucopolysaccharidosis: characterization by MRI. AB - Mucopolysaccharidosis (MPS) type I (alpha-iduronidase deficiency) is characterized by storage and massive urinary excretion of dermatan sulfate and heparan sulfate; it may be distinguished into three different subtypes based on age at onset and severity of the clinical symptoms. We report on progressive white matter involvement documented by serial MR imaging in a patient with the MPS type I, severe skeletal involvement and preserved mental capabilities (intermediate phenotype or Hurler/Scheie syndrome).The natural history of white matter abnormalities in patients with MPS is still unclear; based on the present study, it appears that degenerative changes of the white matter mimicking a leukodystrophy may mark the course of MPS type I. We also suggest that the degree of MR changes in patients with MPS does not always reflect their neurological impairment. PMID- 11897251 TI - The reproducibility of power spectrum analysis of heart rate variability before and after a standardized meal. AB - We tested the reproducibility of the heart rate variability (HRV) measurements before and after a standardized meal. Heart rate recordings were obtained in 14 healthy subjects tested in a reclining position before and after a standardized meal on two separated occasions, apart by a 1-2-week interval. We measured three components of HRV: sympathetic activity (SYMP), parasympathetic activity (PSYMP) and the ratio of SYMP/PSYMP under controlled breathing and noncontrolled breathing conditions. We observed that all components were reproducible during noncontrolled breathing condition, whereas only PSYMP was reproducible during controlled breathing condition. Our results thus indicate that HRV measurements could be a useful, noninvasive and nonexpensive method to provide SYMP and SYMP/PSYMP in feeding behavior studies when measured under noncontrolled breathing conditions. Nonetheless, using a controlled breathing condition may be relevant when assessing the effect of various interventions or drugs on parasympathetic activity. PMID- 11897252 TI - Effect of the 5-HT(1A) receptor agonist 8-OH-DPAT on food and water intake in chickens. AB - The effects of the 5-HT(1A) receptor agonist 8-OH-DPAT on food and water intake in 16-week-old male chickens were investigated. Injection of 25 or 50 microg/kg of 8-OH-DPAT 15 min before refeeding starved animals (starved-refed) produced a decrease in food intake 1 h after the start of refeeding. No effect was observed in water intake. The injection of 25 or 50 microg/kg of the 8-OH-DPAT 60 min after the start of refeeding (fed) produced increased food intake, but no effect was observed on water intake. The agonist 8-OH-DPAT (50 microg/kg) injected in water-deprived chickens 15 min before water presentation produced a rapid increase in water intake and an increase in food intake 90 min after the presentation of water. The effect on food intake was mainly apparent 60-90 min after injection. However, when the chickens were water-deprived, the intravenous administration of 8-OH-DPAT 15 min before the presentation of water produced an increase in water intake only 15 min after the start of the experiment. The results show that the effect on food intake of the agonist 8-OH-DPAT in chickens was similar to that observed in mammals. Also, the results show that the agonist induced increase in water intake may act via a different mechanism. The results show that the 8-OH-DPAT, as in mammals, has a complex effect on food and water intake in chickens and that further works need to be carried out to understand the mechanisms involved in the food and water intake using different animal models. PMID- 11897253 TI - Social stress in male mice impairs long-term antiviral immunity selectively in wounded subjects. AB - An important property of the antiviral immune response is its time-dependent character. Beginning with a few antigen-specific cells upon infection, it evolves to a stage where there is an abundance of antigen-specific cells and antibodies that are needed to clear the pathogen, and ends with circulating antibodies and a population of virus-specific memory cells to protect the animal from reinfection. Short-term effects of stress on the immune system have been investigated extensively, showing that stress acutely changes many aspects of immunity. However, relatively little is known about the consequences of stress for the quality and quantity of long-term immunological memory. In the present study, we have investigated the effect of social stress, applied in mice at Days 1, 2 and 3 after inoculation with a herpes virus, on long-term antibody and memory cytokine responses to the virus. Male mice were subjected to three 5-min confrontations with an aggressive conspecific. Approximately half of the mice was wounded by bites of the aggressor during this stress procedure, and these mice were analyzed separately from nonwounded mice. It appeared that wounded mice showed suppressed protective antibody responses and impaired memory for virus-specific IL-4 and IL 10 production, whereas mice that were not wounded showed intact long-term immune responses and memory. It is concluded that the combination of wounds and the social stress of repeated confrontations is associated with impaired protective immunity as a consequence of suppressed antibody levels and impairment of some aspects of antiviral immunological memory. PMID- 11897254 TI - Testosterone rapidly affects the expression of copulatory behavior in house mice (Mus musculus). AB - Male mammals reflexively release an endogenous pulse of testosterone in response to either a female or her urinary pheromones. Two experiments examined the hypothesis that such pulses have quick-acting effects upon the expression of reproductive behavior in male house mice. In Experiment 1, 30 min after exposure to female urine, males exhibited significantly reduced latency to mount a receptive female (when they should have been expressing an endogenous testosterone pulse). In Experiment 2, gonadally intact males received a simulated testosterone pulse via a subcutaneous injection of 500 microg of testosterone propionate. At 60 min after injection, males mounted a receptive female significantly more quickly than if they had not received such an injection. These experiments provide evidence that elevations in testosterone titers above baseline can, under certain conditions, rapidly alter the expression of male typical behaviors. PMID- 11897255 TI - Evidence that appetitive responses for dehydration and food-deprivation are learned. AB - Rats do not seek water when cellularly dehydrated until they are about 4 weeks of age. This lack of appetitive 'seeking' behavior in young rats differs from their precocious ingestive responses such as an increased intake of solutions infused into their mouths when they are dehydrated as young as 2 days of age. Using video analysis of appetitive behavior in a structured environment, we document this early absence of appetitive responding and the subsequent acquisition of dehydration-elicited appetitive behavior. Weaning age pups were separated into four conditions: (i) experienced, dehydrated; (ii) experienced, nondehydrated; (iii) inexperienced, dehydrated; and (iv) inexperienced, nondehydrated. 'Experienced' rats received a dehydration and drinking experience prior to the test, and 'dehydrated' rats were dehydrated (by injection of a salt load) at the time of test. At the test, all water and food was removed from the test cages, eliminating the confounding of appetitive and consummatory measures. Despite the fact that pups in all conditions had experience with water and had previously drunk, only the 'experienced' pups differentially sought water when dehydrated. Parallel experiments with food deprivation produced similar results. Pups did not exhibit food-seeking behavior when food-deprived unless they had previous experience with food deprivation and eating. The appetitive 'seeking' behavior for feeding also appears to be learned. Directed appetitive behavior in general may thus be acquired. PMID- 11897256 TI - Influence of genetic taste sensitivity to 6-n-propylthiouracil (PROP), dietary restraint and disinhibition on body mass index in middle-aged women. AB - A previous study from our laboratory reported a small, inverse association between taste sensitivity to 6-n-propylthiouracil (PROP) and body mass index (BMI) among lean, young-adult males. Weaknesses in study design precluded making this same observation in females. To overcome these shortcomings, the present study investigated this relationship in older, heavier women characterized by dietary restraint and disinhibition. We tested the hypothesis that lower taste sensitivity to PROP would be associated with higher BMI but that high dietary restraint would mask this association. Thus, no relationship between taster status and BMI was expected in those with high dietary restraint. Eighty-six, middle-aged women (39.0+/-1.0 years) with a mean BMI of 27.4+/-0.8 participated. They were classified as PROP nontasters (n=23), medium tasters (n=32) or supertasters (n=31). Initial analyses revealed that disinhibition had a strong positive influence on BMI that was unrelated to PROP status (P< or =.001). After adjusting for disinhibition, a strong interaction between PROP status and restraint on BMI was noted. Among women with low dietary restraint, nontasters (P< or =.01) and medium tasters (P< or =.05) were heavier than supertasters by 6 and 4 adjusted BMI units, respectively. No differences in BMI were found across taster groups in women with high dietary restraint. These data confirm that the inverse association between PROP status and BMI reported earlier in men is also present in women and that this relationship becomes apparent when variables relevant to eating behavior in women are taken into consideration. These findings are discussed in the context of emerging theories relating genetic differences in taste to fat acceptance, fat intake and body weight. PMID- 11897257 TI - Intake and hedonics of calcium and sodium during pregnancy and lactation in the rat. AB - These experiments sought to distinguish whether increased calcium intake during pregnancy and lactation in the rat is due to arousal of a specific calcium appetite, with altered taste hedonics, as occurs with sodium depletion, to reduced taste sensitivity, or to the hyperdipsia of reproduction. We find that, during pregnancy and lactation, CaCl(2) intake is not increased more (in fact less) than intakes of control tastants, MgCl(2) and quinine HCl, and multiparous dams do not have a greater calcium intake than primaparous dams. Changes in taste reactivity to CaCl(2) and to NaCl do not correlate with changes in intake of these minerals during pregnancy or lactation, suggesting that alterations in hedonics or sensitivity do not explain the increased intake of these minerals. Taken together with the increased intake of all the tastants, it may be that the increased intakes of calcium and sodium during reproduction are not due to respective specific appetites or to a general mineral appetite but rather to the reproduction-increased ingestion that may meet all the dam's increased mineral and nutrient requirements. Differences in the degree of increased intakes of tastes may be due to specific alterations in their transduction during reproduction. PMID- 11897258 TI - Megestrol acetate increases short-term food intake in zinc-deficient rats. AB - Rats offered a zinc-deficient (-Zn) diet voluntarily reduce their food intake within 3-4 days. Megestrol acetate (MA) is an appetite-stimulating drug used to treat cachexia of chronic diseases. In previous work, we found MA administration to male rats increased consumption of a -Zn diet. This approach would provide a useful tool for nutritional studies in which nutrient intake, except for zinc, would be maintained. The present study further examined the use of MA to increase consumption of a -Zn diet over a longer time period in both male and female rats. Rats were fed either a -Zn or a zinc-adequate (+Zn) diet. In Experiment 1, rats were treated orally with 0, 20, 50 or 100 mg MA/kg BW in corn oil for 21 days. MA stimulated intake of the -Zn diet in a linear manner. In Experiments 2 and 3, male and female rats, respectively, were fed the -Zn or +Zn diets and treated with 100 mg MA/kg BW for 21 days. In both experiments, MA administration increased intake of the -Zn diet to levels similar to the +Zn diet through Day 14. MA increased the hypothalamic neuropeptide Y (NPY) concentration in male rats, but did not affect serum IGF-I. MA administration improved growth of female but not male rats fed the -Zn diet. In females, serum IGF-I was not lower in zinc deficient rats, which may have allowed the improved growth response with MA. Hence, MA administration may be a useful tool to increase consumption of a -Zn diet in short-term studies. PMID- 11897259 TI - Efficacy of a 'functional energy drink' in counteracting driver sleepiness. AB - Driver sleepiness is a major cause of serious road crashes. Coffee is often used as an effective countermeasure to driver sleepiness. However, the caffeine levels in coffee are variable, whereas certain proprietary "functional energy drinks" (FEDs) contain known levels of caffeine (and other ingredients). We investigated the effectiveness of a well-known FED in reducing sleepiness in drivers. Twelve healthy young adults drove an instrumented car simulator between 14:00 and 17:00 h. Their sleepiness was enhanced by sleep restriction to 5 h the night before. Following a pretreatment 30-min drive and at the beginning of a 30-min break, participants were given double-blind 250-ml FED (containing sucrose, glucose, 80 mg caffeine, taurine, glucuronolactone and vitamins) vs. a control drink with the same volume and same taste but without caffeine, taurine and glucuronolactone. Two hours of continuous driving ensued. Lane drifting, subjective sleepiness and the electroencephalogram (EEG) were monitored throughout. Compared with the control, the FED significantly reduced sleep-related driving incidents and subjective sleepiness for the first 90 min of the drive. There was a trend for the EEG to reflect less sleepiness during this period. It was concluded that the FED is beneficial in reducing sleepiness and sleep-related driving incidents under conditions of afternoon monotonous driving following sleep restriction the night before. PMID- 11897260 TI - Effect of prenatal androgen receptor antagonist or aromatase inhibitor on sexual behavior, partner preference and neuronal Fos responses to estrous female odors in the rat accessory olfactory system. AB - Many socially relevant odors are detected in rodent species by the vomeronasal organ and subsequently processed by the accessory olfactory system (AOS). We previously found that gonadectomized male and female rats treated in adulthood with testosterone propionate (TP) showed equivalent Fos responses in the AOS to odors derived from estrous females. Likewise, in contrast with numerous other mammalian species, gonadectomized female rats show surprisingly high levels of male-typical mounting behavior in response to adult TP. We tested the hypothesis that prenatal testosterone (T) exposure, acting via androgen receptors (ARs) or via estrogen receptors, masculinizes the AOS in rats of both sexes. Pregnant dams were treated with either the AR blocker, Flutamide, the aromatase inhibitor, 1,4,6-androstatriene-3,17-dione (ATD), or nothing (control) to assess the role of prenatal androgen and estradiol receptor activation, respectively, in this masculinization. Beginning at birth, male and female offspring were injected subcutaneously (sc) every other day with either ATD (pre- and neonatal ATD group) or oil vehicle (Flutamide and control groups) until postnatal Day 12. Subjects were gonadectomized as adults, hormonally treated and tested for different behaviors before having their AOS Fos responses to estrous female odors assessed. Prenatal treatment with Flutamide (but not ATD) significantly decreased anogenital distance and severely impaired intromissive and ejaculatory behaviors in males tested after TP replacement without disrupting mounting capacity in either sex. Pre- and neonatal treatment with ATD (but not Flutamide) enhanced lordosis responsiveness in males tested after sc injections of estradiol and progesterone, whereas these perinatal treatments had no effect on any aspect of masculine coital performance in either sex. After TP treatment, male and female control subjects preferred to approach a tethered stimulus female as opposed to a male, and prenatal Flutamide or perinatal ATD treatments did not modify this pattern of partner preference. Neuronal Fos responses to estrous odors were (as in previous studies) identical in the AOS of gonadectomized TP-treated control males and females. Prenatal Flutamide or perinatal ATD treatments failed to disrupt consistently this profile of Fos responses to estrous odors in the AOS of rats of either sex. These behavioral and neuroanatomical findings raise the possibility that the similar level of male-typical responsiveness to social odors that occurs in male and female rats after adult TP treatment results from nonsteroid-hormone-dependent, species-specific factors that act perinatally in the brains of rats of both sexes. PMID- 11897261 TI - Dissociation between spontaneously emitted and apomorphine-induced stereotypy in Peromyscus maniculatus bairdii. AB - Stereotyped behavior is repetitive, topographically invariant motor activity that lacks an obvious function. We have previously characterized the spontaneous and persistent stereotypies that occur in deer mice housed in standard laboratory cages. Providing these animals with enriched environments markedly reduces their vulnerability to develop stereotypic behavioral repertoires, thus enabling us to generate behaviorally distinct (stereotypic and nonstereotypic) mice of the same species. As stereotypic behaviors are readily induced by systemic administration of a dopamine (DA) agonist, the present study tested whether apomorphine would induce stereotypies in environmentally enriched (nonstereotypic) deer mice that were topographically similar to the stereotypies that are spontaneously emitted by standard-caged (stereotypic) deer mice. The effects of apomorphine were also evaluated in the standard-caged (stereotypic) deer mice. DA agonist-induced behaviors in nonstereotypic mice included stereotypies that were largely topographically distinct from spontaneously emitted stereotypies; apomorphine failed to produce statistically significant elevations in two of the three stereotypic behaviors typical of standard-caged deer mice. Furthermore, there was no evidence of increased DA receptor sensitivity in stereotypic mice. Thus, environmentally related stereotypy is distinct from systemically administered DA agonist-induced stereotypy, and is not exacerbated by such drug treatment. The results obtained do provide support, however, for a limited involvement of the DA system in the mediation of these behaviors. PMID- 11897263 TI - Chronic oral administration of clomipramine decreases sexual behavior in the male Syrian hamster (Mesocricetus auratus). AB - Previous studies have demonstrated that clomipramine (CLI), a selective serotonin reuptake inhibitor, has negative side effects on sexual behavior when administered to adult rats and humans. In addition, neonatal rat pups given chronic doses of CLI display similar sexual deficits upon sexual maturity. This study used two experiments to test the hypothesis that 2 weeks of CLI would cause a decrease in sexual motivation and performance in the male Syrian hamster (Mesocricetus auratus). Experiment 1 administered 0, 40, or 60 mg/kg CLI for 14 days to adult male hamsters via a sugar water solution. Experiment 2 administered 0, 40, or 60 mg/kg CLI for 14 days to pregnant dams during gestation (also via a sugar water solution). We hypothesized that this administration of CLI via the pregnant mother would have long-lasting developmental effects on the male hamster pups resulting in later dysfunction in male sexual behavior. Results from Experiment 1 indicate that CLI administration to adult males caused a significant decrease in the number of ejaculations compared to controls. The only significant difference in sexual behavior between those males whose mothers received CLI during pregnancy compared to males from untreated mothers was an increase in the number of intromissions in the highest dosage group. These findings demonstrate that CLI inhibits sexual performance in adult male Syrian hamsters, and also suggest that oral administration of CLI via a sugar water solution is an effective mode of administration. PMID- 11897262 TI - Alleviating effects of plant-derived fragrances on stress-induced hyperthermia in rats. AB - In the present study, the effects of exposure to plant-derived odors on the autonomic and behavioral responses to novel environment were examined in rats. Male rats (n=42) carrying a telemetry transmitter were individually housed, and on the test day each rat was transferred to a new cage containing bedding that had been sprayed immediately before testing with 200 microl of 0.03% dilution of either lavender essential oil, green leaf odor (a mixture of hexenol and hexenal), alpha-pinene, or solvent (triethyl citrate) as a control. Following the transfer to this novel environment, the body temperature of the rats increased by nearly 1 degrees C, showing a stress-induced hyperthermia. Stress-induced hyperthermia was attenuated by the green odor and the alpha-pinene, but not by the lavender or solvent. There was no clear effect of fragrances on heart rate or behavioral responses. These results suggest that plant-derived fragrances, such as green odor and alpha-pinene, have calming effects on autonomic stress response to novel environments. PMID- 11897264 TI - Pheromonal influences on sociosexual behavior in young women. AB - A double-blind, placebo-controlled study of a synthesized putative female pheromone was conducted with regularly menstruating, university women (N=36, mean age=27.8). The pheromone formula was derived from earlier work investigating the underarm secretions of fertile, sexually active, heterosexual women. A vial of either synthesized pheromone or placebo was selected blindly and added to a subject's perfume. Subjects recorded seven sociosexual behaviors and reported them weekly across three menstrual cycles. Beginning with Day 8 of each cycle, the first cycle contained a 2-week baseline period followed by an experimental period of as many as 3 weeks each from the next two cycles for a maximum of 6 weeks. The 19 pheromone and 17 placebo subjects did not differ significantly in age, weight, body mass index, dating status or ethnicity nor in reported accuracy, back-filling data, perception of a positive effect or perfume use. Placebo subjects were significantly taller than pheromone subjects. Except for male approaches, subjects did not differ significantly at baseline in average weekly sociosexual behaviors. A significantly greater proportion of pheromone users compared with placebo users increased over baseline in frequency of sexual intercourse, sleeping next to a partner, formal dates and petting/affection/kissing but not in frequency of male approaches, informal dates or masturbation. Three or more sociosexual behaviors increased over baseline for 74% of pheromone users compared with 23% of placebo users. We conclude that this synthesized pheromone formula acted as a sex attractant pheromone and increased the sexual attractiveness of women to men. PMID- 11897265 TI - Electrophysiological analysis of rhythmic jaw movements in the freely moving mouse. AB - Although rhythmic jaw movement in feeding has been studied in mammals, such as rats, rabbits and monkeys, the cellular and molecular mechanisms underlying it are not well understood. Transgenic and gene-targeting technologies enable direct control of the genetic makeup of the mouse, and have led to the development of a new category of reagents that have the potential to elucidate the cellular and molecular mechanisms of neural networks. The present study attempts to characterize rhythmic jaw movements in the mouse and to demonstrate its relevance to rhythmic jaw movements found in higher mammals using newly developed jaw tracking systems and electromyograms of the masticatory muscles. The masticatory sequence of the mouse during feeding was classified into two stages, incision and chewing. Small and rapid (8 Hz) open-close jaw movements were observed during incision, while large and slow (5 Hz) open-close jaw movements were observed during chewing. Integrated electromyograms of the masseteric and digastric muscles were larger during chewing than those observed during incision. Licking behavior was associated with regular (8 Hz), small open-close jaw movements with smaller masseteric activity than those observed during mastication. Grooming showed variable patterns of jaw movement and electromyograms depending on the grooming site. These results suggest that there are neuronal mechanisms producing different frequencies of rhythmic jaw movements in the mouse, and we conclude that the mouse is useful for understanding rhythmic jaw movements in higher mammals. PMID- 11897266 TI - Conditioned taste aversion using four different means to deliver sucrose to rats. AB - A solution of sucrose either to be drunk from a drinking tube-self-drinking procedure (SD)-or perfused intraorally as a consequence of nose-pokes-self administration procedure (SA)-or perfused as a consequence of licking an empty tube (LA)-was paired with an LiCl-induced malaise in rats. The effects were compared to those of a procedure consisting of intraoral administration (IO) of sucrose not contingent to any specific action of the rat. Similar levels of conditioned taste aversion (CTA) were obtained but extinction in the IO procedure was quicker than in the SA procedure, which was itself quicker than in the SD procedure. Extinctions in the IO and LA procedures resembled one another and were quicker than in the SD procedure. A step towards deciding between several explanatory hypotheses of these differences was made by conducting two more experiments. The third experiment was based on reinstatement, or not, of the conditioning procedure for the test after standard IO extinction. CTA was produced only when SD was used both at conditioning and test. A fourth experiment was based on latent inhibition where the procedure was changed, or not, between preexposure and conditioning. Latent inhibition was absent only when the rats had been preexposed to sucrose with the SA procedure and conditioned with the SD procedure. PMID- 11897267 TI - Responsivity to NPY and melanocortins in obese OLETF rats lacking CCK-A receptors. AB - Otsuka Long-Evans Tokushima Fatty (OLETF) rat lacking CCK-A receptors are hyperphagic and obese. Previous work has demonstrated alterations in neuropeptide Y (NPY) and proopiomelanocortin (POMC) mRNA expression in ad libitum fed OLETF rats compared to lean Long-Evans Tokushima Otsuka (LETO) controls. In order to determine whether alterations in sensitivity to central peptides involved in overall feeding control may contribute to the hyperphagia and obesity in OLETF rats, we assessed OLETF and LETO rats feeding responses to lateral ventricular infusions of NPY (1 and 3.2 nmol), the melanocortin 3/4 agonist MTII (0.1 and 0.32 nmol) and the melanocortin receptor antagonist SHU-9119 (0.25 and 0.5 nmol). At a 3-h time point, NPY increased food intake in both OLETF and LETO rats. OLETF rats were more sensitive, having significant increases at both NPY doses and a greater increase at the higher dose. The melanocortin agonist MTII decreased intake in both LETO and OLETF rats. At the 20-h time point, the magnitude of suppression was greater in OLETF rats. SHU-9119 increased food intake in both groups. OLETF rats were more sensitive with larger relative increase and longer lasting effects at the lower dose. These results are consistent with demonstrated alterations in neuropeptide gene expression in OLETF rats and indicate that alterations in responsivity to NPY and melanocortin signaling are unlikely to contribute to their hyperphagia and obesity. PMID- 11897268 TI - Behavioral and magnetic resonance spectroscopic studies in the rat hyperserotonemic model of autism. AB - Autism is classified as a pervasive developmental disorder, with several cardinal features including sensory disturbances, obsessive-compulsive-like behavior, lack of bonding to caregivers and motor disturbances. To date, there is a lack of an animal model of the disease. The current work is aimed at producing such a model by treating developing rat pups with a serotonergic agonist, 5-methoxytryptamine (5-MT; 1 mg/kg) during development (from gestational age 12 days to postnatal day 20), thus mimicking one of the hallmark neurochemical features of the illness increases in the neurotransmitter, serotonin. Animals were then tested in behavioral paradigms that may resemble the human illness. Treated rat pups were found to be overreactive to auditory or tactile sensory stimuli, to display changes in the negative geotaxic test of motor development, to show lack of separation-induced vocalizations when their dam was removed and to show decreased alternation in the spontaneous alternation task. As well, the animals showed metabolic abnormalities in the brain using in vivo proton magnetic resonance spectroscopy, which are consistent with those observed in autistic children. In summary, the model we are proposing shows some of the behavioral and metabolic features of autism, as well as being produced through alteration of a neurochemical system known to be altered in autism. PMID- 11897269 TI - Carbohydrate to protein ratio in food and cognitive performance in the morning. AB - The effect of different carbohydrate to protein ratios in food on cognitive functions and the relation between postprandial metabolic and cognitive changes were studied in 15 healthy male students. Subjects were tested in three sessions, separated by 1 week, for short-term changes in mood states, objective cognitive functions, blood parameters, and indirect calorimetry using a repeated-measures, counterbalanced cross-over design. Measurements were made after an overnight fast before and hourly during 3.5 h after test meal ingestion. The isoenergetic (1670 kJ) test meals consisted of three carbohydrate to protein ratios, i.e. a carbohydrate-rich (CHO[4:1]), balanced (BAL[1:1]), and protein-rich (PRO[1:4]) meal, respectively. Overall accuracy in short-term memory was best after the PRO[1:4] meal concomitant to the least variation in glucose metabolism and glucagon to insulin ratio (GIR). Related to changes in glucose metabolism and/or in the ratios of large neutral amino acids (LNAA), respectively, attention and decision times were transiently improved within the first hour after the CHO[4:1] meal, whereas after the first hour the BAL[1:1] and PRO[1:4] meal resulted in improved performance. Overall reaction times of a central task were fastest after the BAL[1:1] meal concomitant to the highest overall tyrosine (Tyr) to LNAA ratio. Our findings suggest that the carbohydrate to protein ratio in food specifically influences higher cognitive functions in the morning. Except for a transient positive effect of rising blood glucose after a carbohydrate-rich meal, a protein-rich or balanced meal seems to result in better overall cognitive performance presumably because of less variation in glucose metabolism and/or higher modulation in LNAA ratios indicated by the overall GIR. PMID- 11897270 TI - Increased hypothalamic neuropeptide Y expression in deprived preweanling rats is reversed by intragastric infusion of milk. AB - Lean, preweanling Zucker rat pups increase neuropeptide Y (NPY) expression in the hypothalamic arcuate nucleus in response to a 24-h deprivation of food, water, and maternal interaction as early as postnatal day 2 (P2). In this study, we examined if replacing nutritive or tactile aspects of maternal behavior to deprived rat pups could block the increased expression of hypothalamic NPY measured by in situ hybridization. On P2, P12, or P15, littermates were assigned to one of four treatment groups: (1) left with the dam for 24 h, (2) deprived of the dam for 24 h and given tactile stimulation in the form of periodic anogenital stroking to elicit urination and defecation, (3) deprived of the dam and given periodic anogenital stroking plus continuous gastric infusion of milk for 24 h, or (4) deprived of the dam and given periodic anogenital stroking plus continuous infusion of water for 24 h. We found that gastric infusions of milk normalized NPY expression at all three ages, gastric infusions of water did not on P2 and P15, and anogenital stroking alone had no effect. We suggest that the lack of milk is the major cause of increased hypothalamic NPY expression during maternal deprivation in lean Zucker pups. PMID- 11897272 TI - Room at the top. PMID- 11897273 TI - Simian virus 40 and non-Hodgkin lymphoma. PMID- 11897274 TI - Addressing suicide as a public-health problem. PMID- 11897275 TI - Partnerships between expert patients and physicians. PMID- 11897276 TI - Management of oral mucositis associated with cancer chemotherapy. PMID- 11897278 TI - Association between simian virus 40 and non-Hodgkin lymphoma. AB - BACKGROUND: Non-Hodgkin lymphoma has increased in frequency over the past 30 years, and is a common cancer in HIV-1-infected patients. Although no definite risk factors have emerged, a viral cause has been postulated. Polyomaviruses are known to infect human beings and to induce tumours in laboratory animals. We aimed to identify which one of the three polyomaviruses able to infect human beings (simian virus 40 [SV40], JC virus, and BK virus) was associated with non Hodgkin lymphoma. METHODS: We analysed systemic non-Hodgkin lymphoma from 76 HIV 1-infected and 78 HIV-1-uninfected patients, and non-malignant lymphoid samples from 79 HIV-1-positive and 107 HIV-1-negative patients without tumours; 54 colon and breast carcinoma samples served as cancer controls. We used PCR followed by Southern blot hybridisation and DNA sequence analysis to detect DNAs of polyomaviruses and herpesviruses. FINDINGS: Polyomavirus T antigen sequences, all of which were SV40-specific, were detected in 64 (42%) of 154 non-Hodgkin lymphomas, none of 186 non-malignant lymphoid samples, and none of 54 control cancers. This difference was similar for HIV-1-infected patients and HIV-1 uninfected patients alike. Few tumours were positive for both SV40 and Epstein Barr virus. Human herpesvirus type 8 was not detected. SV40 sequences were found most frequently in diffuse large B-cell and follicular-type lymphomas. INTERPRETATION: SV40 is significantly associated with some types of non-Hodgkin lymphoma. These results add lymphomas to the types of human cancers associated with SV40. PMID- 11897279 TI - Boerhaave's syndrome. PMID- 11897280 TI - Effect of 6-week course of glucagon-like peptide 1 on glycaemic control, insulin sensitivity, and beta-cell function in type 2 diabetes: a parallel-group study. AB - BACKGROUND: Glucagon-like peptide 1 (GLP-1) has been proposed as a treatment for type 2 diabetes. We have investigated the long-term effects of continuous administration of this peptide hormone in a 6-week pilot study. METHODS: 20 patients with type 2 diabetes were alternately assigned continuous subcutaneous infusion of GLP-1 (n=10) or saline (n=10) for 6 weeks. Before (week 0) and at weeks 1 and 6, they underwent beta-cell function tests (hyperglycaemic clamps), 8 h profiles of plasma glucose, insulin, C-peptide, glucagon, and free fatty acids, and appetite and side-effect ratings on 100 mm visual analogue scales; at weeks 0 and 6 they also underwent dexascanning, measurement of insulin sensitivity (hyperinsulinaemic euglycaemic clamps), haemoglobin A(1c), and fructosamine. The primary endpoints were haemoglobin A(1c) concentration, 8-h profile of glucose concentration in plasma, and beta-cell function (defined as the first-phase response to glucose and the maximum insulin secretory capacity of the cell). Analyses were per protocol. FINDINGS: One patient assigned saline was excluded because no veins were accessible. In the remaining nine patients in that group, no significant changes were observed except an increase in fructosamine concentration (p=0.0004). In the GLP-1 group, fasting and 8 h mean plasma glucose decreased by 4.3 mmol/L and 5.5 mmol/L (p<0.0001). Haemoglobin A(1c) decreased by 1.3% (p=0.003) and fructosamine fell to normal values (p=0.0002). Fasting and 8 h mean concentrations of free fatty acids decreased by 30% and 23% (p=0.0005 and 0.01, respectively). Gastric emptying was inhibited, bodyweight decreased by 1.9 kg, and appetite was reduced. Both insulin sensitivity and beta-cell function improved (p=0.003 and p=0.003, respectively). No important side-effects were seen. INTERPRETATION: GLP-1 could be a new treatment for type 2 diabetes, though further investigation of the long-term effects of GLP-1 is needed. PMID- 11897281 TI - Frequency, severity, and duration of rhinovirus infections in asthmatic and non asthmatic individuals: a longitudinal cohort study. AB - BACKGROUND: Rhinovirus infections cause exacerbations of asthma. We postulated that people with asthma are more susceptible to rhinovirus infection than people without the disease and compared the susceptibility of these groups. METHODS: We recruited 76 cohabiting couples. One person in every couple had atopic asthma and one was healthy. Participants completed daily diary cards of upper-respiratory tract (URT) and lower-respiratory-tract (LRT) symptoms and measured peak expiratory flow twice daily. Every 2 weeks nasal aspirates were taken and examined for rhinovirus. Mixed models were used to compare risks of infection between groups. We also compared the severity and duration of infections. FINDINGS: We analysed 753 samples. Rhinovirus was detected in 10.1% (38/378) of samples from participants with asthma and 8.5% (32/375) of samples from healthy participants. After adjustment for confounding factors, asthma did not significantly increase risk of infection (odds ratio 1.15, 95% CI 0.71-1.87). Groups did not differ in frequency, severity, or duration of URT infections or symptoms associated with rhinovirus infection. First rhinovirus infection was associated more frequently with LRT infection in participants with asthma than in healthy individuals (12 of 28 infections vs four of 23, respectively, p=0.051). Symptoms of LRT associated with rhinovirus infection were significantly more severe (p=0.001) and longer-lasting in participants with asthma than in healthy participants (p=0.005). INTERPRETATION: People with atopic asthma are not at greater risk of rhinovirus infection than healthy individuals but suffer from more frequent LRT infections and have more severe and longer-lasting LRT symptoms. PMID- 11897282 TI - Uses of error: the deceitful heart. PMID- 11897283 TI - Suicide rates in China, 1995-99. AB - BACKGROUND: A wide range of suicide rates are reported for China because official mortality data are based on an unrepresentative sample and because different reports adjust crude rates in different ways. We aimed to present an accurate picture of the current pattern of suicide in China on the basis of conservative estimates of suicide rates in different population cohorts. METHODS: Suicide rates by sex, 5-year age-group, and region (urban or rural) reported in mortality data for 1995-99 provided by the Chinese Ministry of Health were adjusted according to an estimated rate of unreported deaths and projected to the corresponding population. FINDINGS: We estimated a mean annual suicide rate of 23 per 100,000 and a total of 287,000 suicide deaths per year. Suicide accounted for 3(.)6% of all deaths in China and was the fifth most important cause of death. Among young adults 15-34 years of age, suicide was the leading cause of death, accounting for 19% of all deaths. The rate in women was 25% higher than in men, mainly because of the large number of suicides in young rural women. Rural rates were three times higher than urban rates-a difference that remained true for both sexes, for all age-groups, and over time. INTERPRETATION: Suicide is a major public-health problem for China that is only gradually being recognised. The unique pattern of suicides in China is widely acknowledged, so controversy about the overall suicide rate should not delay the development and testing of China specific suicide-prevention programmes. PMID- 11897284 TI - Autosomal recessive hypercholesterolaemia in Sardinia, Italy, and mutations in ARH: a clinical and molecular genetic analysis. AB - BACKGROUND: Autosomal recessive hypercholesterolaemia (ARH) is caused by mutations in a putative adaptor protein called ARH. This recessive disorder, characterised by severe hypercholesterolaemia, xanthomatosis, and premature coronary artery disease, is rare except on the island of Sardinia, Italy. Our aim was to ascertain why ARH is more common on Sardinia than elsewhere. METHODS: We obtained detailed medical histories, did physical examinations, measured concentrations of lipoproteins, and harvested genomic DNA from 28 Sardinians with ARH from 17 unrelated families. We sequenced the coding regions and consensus splice sites of ARH in probands from these families, and from 40 individuals of non-Sardinian origin who had an autosomal recessive form of hypercholesterolaemia of unknown cause. FINDINGS: Two ARH mutations, a frameshift mutation (c432insA) in exon 4 (ARH1) and a nonsense mutation (c65G-->A) in exon 1 (ARH2), were present in all of the 17 unrelated families with ARH. Three of the ARH alleles contained both mutations, as a result of an ancient recombination between ARH1 and ARH2. No regional clustering of the three mutant alleles within Sardinia was apparent. Furthermore, four Italians from the mainland with autosomal recessive hypercholesterolaemia were homozygous for ARH1. INTERPRETATION: The small number, high frequency, and dispersed distribution of ARH mutations on Sardinia are consistent with these mutations being ancient and maintained in the Sardinian population because of geographic isolation. PMID- 11897285 TI - A race to the lab. PMID- 11897286 TI - Bioavailability of subcutaneous low-molecular-weight heparin to patients on vasopressors. AB - Venous thromboembolism is a frequent complication in patients admitted to intensive care units (ICU), despite prophylactic treatment with subcutaneous low molecular-weight (LMW) heparin. We postulated that poor efficacy of subcutaneous heparin might be due to administration of vasopressors, which could cause impaired peripheral circulation and inadequate systemic bioavailability of the anticoagulant. We compared concentrations of factor Xa activity in three groups of 15 patients: individuals in ICU who had and had not received vasopressors, and general surgery patients. Those who received vasopressors had lower plasma concentrations of factor-Xa activity than patients in ICU not on vasopressors and postoperative controls. Patients in ICU who take vasopressors could need higher doses of LMW heparin, or a different mode of administration of the drug, to attain adequate thrombosis prophylaxis. PMID- 11897287 TI - Presence of simian virus 40 DNA sequences in human lymphomas. AB - Simian virus 40 (SV40)--a potent oncogenic virus--has been associated previously with some types of human tumours, but not with lymphomas. We examined human tumours for the presence of specific SV40 DNA sequences by PCR and Southern blotting. Viral sequences were present in 29 (43%) of 68 non-Hodgkin lymphomas, and in three (9%) of 31 of Hodgkin's lymphomas. Viral sequences were detected at low frequencies (about 5%) in 235 epithelial tumours of adult and paediatric origin, and were absent in 40 control tissues. Our data suggest that SV40 might be a cofactor in the pathogenesis of non-Hodgkin lymphomas. PMID- 11897288 TI - Fatal allergic vasculitis associated with celecoxib. AB - We report on the occurrence of a rare and as yet unforseeable adverse reaction to treatment with celecoxib, a cyclooxygenase-2 (COX-2) selective, non-steroidal, anti-inflammatory drug. A previously healthy adult suffered fatal acute multiple organ failure presumably after diffuse allergic vasculitis with diffuse necrotic purpura. Although no conclusive proof is available, such a reaction could have been triggered by at least one of two mechanisms: an allergic reaction linked to the chemical structure of celecoxib; or an interaction of the drug with synthesis of endothelial eiconasoids leading to an imbalance between vasoactive end products, resulting in widespread rise to local thrombosis. PMID- 11897289 TI - Provision of information about drug side-effects to patients. AB - To make informed decisions about taking medicinal drugs, people need accurate information about side-effects. A European Union guideline now recommends use of qualitative descriptions for five bands of risk, ranging from very rare (affecting <0.01% of the population), to very common (>10%). We did four studies of more than 750 people, whom we asked to estimate the probability of having a side-effect on the basis of qualitative and quantitative descriptions. Our results showed that qualitative descriptions led to gross overestimation of risk. Until further work is done on how patients taking the drugs interpret these terms, the terms should not be used in drug information leaflets. PMID- 11897290 TI - Confusion over Mandela's "support" of ANC HIV policy. PMID- 11897295 TI - South Asia's governments exhorted to focus on CVD. PMID- 11897296 TI - Facing up to Pakistan's cardiovascular challenge. PMID- 11897297 TI - Report finds abuse in US nursing homes goes unreported and unpunished. PMID- 11897298 TI - Sex-for-food scandal in West African refugee camps. PMID- 11897299 TI - Ghana's new budget draws cheers and jeers in the health sector. PMID- 11897300 TI - Europe consults on more accurate prescribing for children. PMID- 11897302 TI - Arteriovenous malformations. AB - Arteriovenous malformations of the brain are congenital vascular lesions that affect 0.01-0.50% of the population, and are generally present in patients aged 20-40 years. The usual clinical presentations are haemorrhage, seizures, progressive neurological deficit, or headache. Results of natural history studies have shown a yearly haemorrhage rate of 1-4%. Frequency of rebleeding has increased over the years, and several factors that increase risk of haemorrhage have been identified. Although substantial, the morbidity associated with haemorrhages could be less than previously thought. Over the past decade, great advances have been made in application of endovascular embolisation techniques, stereotactic radiosurgery, and microsurgery, allowing effective multidisciplinary treatment of arteriovenous malformations, including those previously deemed to be untreatable. Increasing attention has been paid to management of flow-related aneurysms associated with these malformations. Finally, many reports of recurrent arteriovenous malformations have coincided with new theories regarding the embryogenesis of these disorders and laboratory work suggesting their proliferative potential. PMID- 11897303 TI - Threat of a biological terrorist attack on the US food supply: the CDC perspective. AB - Deliberate contamination of food with biological agents has already been perpetrated in the USA. The US food supply is increasingly characterised by centralised production and wide distribution of products. Deliberate contamination of a commercial food product could cause an outbreak of disease, with many illnesses dispersed over wide geographical areas. Dependent on the biological agent and contaminated food, such an outbreak could either present as a slow, diffuse, and initially unremarkable increase in sporadic cases, or as an explosive epidemic suddenly producing many illnesses. Preparedness for a bioterrorist event affecting the food supply, therefore, entails augmentation of the traditional public-health infrastructure to enhance disease surveillance, accelerate capacity of laboratory detection, rapidly investigate and control outbreaks, and develop capacity for response to mass-casualty disasters. PMID- 11897304 TI - Uses and abuses of screening tests. AB - Screening tests are ubiquitous in contemporary practice, yet the principles of screening are widely misunderstood. Screening is the testing of apparently well people to find those at increased risk of having a disease or disorder. Although an earlier diagnosis generally has intuitive appeal, earlier might not always be better, or worth the cost. Four terms describe the validity of a screening test: sensitivity, specificity, and predictive value of positive and negative results. For tests with continuous variables--eg, blood glucose--sensitivity and specificity are inversely related; where the cutoff for abnormal is placed should indicate the clinical effect of wrong results. The prevalence of disease in a population affects screening test performance: in low-prevalence settings, even very good tests have poor predictive value positives. Hence, knowledge of the approximate prevalence of disease is a prerequisite to interpreting screening test results. Tests are often done in sequence, as is true for syphilis and HIV-1 infection. Lead-time and length biases distort the apparent value of screening programmes; randomised controlled trials are the only way to avoid these biases. Screening can improve health; strong indirect evidence links cervical cytology programmes to declines in cervical cancer mortality. However, inappropriate application or interpretation of screening tests can rob people of their perceived health, initiate harmful diagnostic testing, and squander health-care resources. PMID- 11897305 TI - A brief history of calcitonin. PMID- 11897306 TI - Survival after bone-marrow transplantation. PMID- 11897308 TI - Survival after bone-marrow transplantation. PMID- 11897309 TI - Survival after bone-marrow transplantation. PMID- 11897310 TI - Heroin prescription for opioid addicts. PMID- 11897311 TI - Dynamic epidemiology of group A streptococcal serotypes. PMID- 11897312 TI - Postpartum blindness. PMID- 11897315 TI - Loss of taste is loss of weight. PMID- 11897316 TI - Cerebral venous thrombosis due to protein S deficiency in pregnancy. PMID- 11897317 TI - Economics of drug treatment. PMID- 11897319 TI - Genetics of seasonal affective disorder. PMID- 11897320 TI - Factor V Leiden: relation to fertility? PMID- 11897322 TI - Good medical practice for occupational health. PMID- 11897323 TI - Expanding colonies and expanding repeats. PMID- 11897324 TI - Literature searches. PMID- 11897325 TI - Scientific English for non-English speakers. PMID- 11897327 TI - Victorian children on the wards. PMID- 11897328 TI - Discarded diagnoses. PMID- 11897334 TI - Haunted papers. PMID- 11897336 TI - The fundamental nature of life as a chemical system: the part played by inorganic elements. AB - In this article we show why inorganic metal elements from the environment were an essential part of the origin of living aqueous systems of chemicals in flow. Unavoidably such systems have many closely fixed parameters, related to thermodynamic binding constants, for the interaction of the essential exchangeable inorganic metal elements with both inorganic and organic non-metal materials. The binding constants give rise to fixed free metal ion concentration profiles for different metal ions and ligands in the cytoplasm of all cells closely related to the Irving-Williams series. The amounts of bound elements depend on the organic molecules present as well as these free ion concentrations. This system must have predated coding which is probably only essential for reproductive life. Later evolution in changing chemical environments became based on the development of extra cytoplasmic compartments containing quite different energised free (and bound) element contents but in feed-back communication with the central primitive cytoplasm which changed little. Hence species multiplied late in evolution in large part due to the coupling with the altered inorganic environment. PMID- 11897337 TI - Gold helps bacteria to oxidize methane. AB - With the use of labeled methane-14C and by chromatographic analysis it was shown that gold-containing protein ("Au-protein"), isolated from goldphilic Micrococcus luteus bacteria, catalyzes the oxidation of methane to methanol in the system also containing NADH, air, K(3)Fe(CN)(6) and Tris-HCl buffer. Presumably Au protein helps bacteria to survive when usual sources of carbon and energy are scarce. PMID- 11897338 TI - (Aminoethanol)dichloroplatinum(II) complexes: influence of the hydroxyethyl moiety on 5'-GMP and DNA binding, intramolecular stability, the partition coefficient and anticancer activity. AB - The influence of tethered hydroxyl groups on the binding behavior of the three (aminoethanol)dichloroplatinum complexes, dichloro(N,N'-bis(2 hydroxyethyl)ethylenediamine)-platinum(II) (1), dichloro(N-(2 hydroxyethyl)ethylenediamine)platinum(II) (2) and cis-dichlorobis(2 hydroxyethylamine)platinum(II) (3) towards 5'-GMP and DNA was investigated by 1H NMR and r(b) measurements, respectively. At pH 7.2, the sequence of reactivity with 5'-GMP is 1>2>>3. Complex 3 reacts very slowly with 5'-GMP and DNA and the amount and lifetime of the intermediate 5'-GMP monoadduct are much larger than for 1 and 2. At pH 5.5, the reaction of 3 with 5'-GMP is markedly accelerated and very small amounts of monoadduct are observed, indicating a pH-dependent ability of the pendant hydroxyl group to interact with the platinum moiety. In addition, the effect of the hydroxyethyl functionality on octanol/water partitioning and in vitro anticancer activity was studied. No correlation between lipophilicity and anticancer activity was detected. Furthermore, the lipophilicity and anticancer activity could not be directly correlated to 5'-GMP or DNA binding activity. PMID- 11897339 TI - Studies of the binding of a series of platinum(IV) complexes to plasma proteins. AB - The platinum(IV) complexes: [PtCl(4)(en)], cis,trans-[PtCl(2)(OAc)(2)(en)], cis,trans-[PtCl(2)(OH)(2)(en)] and trans-[Pt(OH)(2)(ethmal)(en)], encompassing a range of reduction potentials and their platinum(II) analogue [PtCl(2)(en)], have been assayed for their protein binding ability in the presence of albumin, albumin and L-cysteine and RPMI 1640 tissue culture medium supplemented with foetal calf serum (RPMI/FCS). cis,trans-[PtCl(4)(en)] exhibited significant protein binding in all three experiments, in a similar fashion to the platinum(II) complex, presumably as a consequence of its rapid reduction. The remaining three platinum(IV) complexes displayed little if any protein binding, with the greatest amount of binding observed in the RPMI/FCS experiment. The extent of binding in the RPMI/FCS correlated with the reduction potentials of the complexes, with the most readily reduced species binding to the greatest extent. PMID- 11897340 TI - Crystal packing and hydrogen bonding in platinum(II) nucleotide complexes: X-ray crystal structure of [Pt(MeSCH(2)CH(2)SMe)(5'-GMP-N7)(2)].6H(2)O. AB - We have synthesised the complex [Pt(CH(3)SCH(2)CH(2)SCH(3))(5'-GMP-N7)(2)].6H(2)O (1), where 5'-GMP is 5'-guanosine monophosphate, and determined its X-ray crystal structure. Pt(II) adopts a square-planar geometry in which the bases are coordinated head-to-tail (HT) in the Delta configuration. The nucleotide conformation in this complex is almost identical to that in the previously reported complex [Pt(en)(5'-GMP-N7)(2)].9H(2)O (2), in which there is outer sphere macrochelation via intramolecular H-bonding between the monoanionic phosphate groups and the coordinated ethylenediamine (en) NH. It is therefore apparent that intermolecular interactions rather than intramolecular H-bonding determines the orientation of the sugar-phosphate side-chain in these Pt(II) bisnucleotide complexes in the solid state. PMID- 11897341 TI - Solution structural studies of molybdate-nucleotide polyanions. AB - The complexation of molybdate with the nucleotides adenosine-5'-monophosphate (5' AMP), adenosine-3'-monophosphate (3'-AMP) and guanosine-5'-monophosphate (5'-GMP) has been investigated by (1)H and (31)P NMR and Mo K-edge X-ray absorption near edge (XANES) and extended X-ray absorption fine structure (EXAFS) spectroscopy. Acidification of aqueous solutions containing molybdate and each of the nucleotides resulted in the formation of a single species characterized by (1)H resonances which are deshielded relative to those of free nucleotide. Analysis of the two-component systems indicated a Mo/nucleotide ratio of 2.5:1 for the complexation species. White compounds, characterized as Na(2)[Mo(5)O(15)(HB)(2)] (B=5'-AMP, 5'-GMP), have been isolated from the acidified molybdate/H(2)B solutions. Dissolution in D(2)O replicates the NMR spectra of the solution species observed prior to precipitation. Solution and solid state Mo K-edge XAS and EXAFS spectroscopy of Na(2)[Mo(5)O(15)(HAMP)(2)] and Na(6)[Mo(5)O(15)(PO(4))(2)] provide convincing evidence for the presence of a pentamolybdodiphosphate core in the molybdate-nucleotide complexes in both the solid and solution states. PMID- 11897342 TI - (Catecholato)iron(III) complexes: structural and functional models for the catechol-bound iron(III) form of catechol dioxygenases. AB - Catechol dioxygenases are mononuclear non-heme iron enzymes that catalyze the oxygenation of catechols to aliphatic acids via the cleavage of aromatic rings. In the last 20 years, a number of (catecholato)iron(III) complexes have been synthesized and characterized as structural and functional models for the catechol-bound iron(III) form of catechol dioxygenases. This review focuses on the structural and spectroscopic characteristics and oxygenation activity of the title complexes. PMID- 11897343 TI - A supramolecular enzyme model catalyzing the central cleavage of carotenoids. AB - Several bis-beta-cyclodextrin porphyrins have been prepared as supramolecular receptors of carotenoids. The binding constants of carotenoids to receptors were determined by quenching the fluorescence of the porphyrins on hydrophobic binding of carotenoids within the cavities of cyclodextrins. K(a)=8.3 x 10(6) M(-1) was calculated for binding of beta,beta-carotene to bis-beta-cyclodextrin Zn porphyrin. The corresponding Ru complex catalyzes the central cleavage of carotenoids in the presence of tert-butyl hydroperoxide in a biphasic system. PMID- 11897344 TI - Aluminum corrolin, a novel chlorophyll analogue. AB - A corrole-based chlorophyll analogue has been prepared, based on the notation that the major differences between the prosthetic groups of chlorophylls and hemes are the presence of a non-transition metal (Mg vs. Fe) and one reduced double bond in the porphyrin ligand. As corroles act as tri- rather than dianionic ligands, the analogy requires the insertion of aluminum into the macrocycle and the reduction of one of its double bonds, two reactions that have not been previously reported with any corrole. The aluminum complexes of both the corrole and the corrolin (the dihydrocorrole) display fluorescence quantum yield that are much larger than of chlorophyll and of all other previously reported synthetic analogues. The results suggest that the light metal atom ion is responsible for low intersystem crossing probability to the triplet excited state and the structural rigidity of the hexa-coordinated complexes for reducing the probability of internal conversion. PMID- 11897345 TI - Coordination geometry of Cu-porphyrin in Cu(II)-Fe(II) hybrid hemoglobins studied by Q-band EPR and resonance Raman spectroscopies. AB - Cu(II)-Fe(II) hybrid hemoglobins were investigated by UV-vis, Q-band (35 GHz) EPR and resonance Raman spectroscopies. EPR results indicated that Cu-porphyrin in alpha-subunit within hybrid hemoglobin had either 5- or 4-coordination geometry depending on the pH conditions, while Cu-porphyrin in beta-subunit had only 5 coordination geometry at high and low pH values. These results were consistent with UV-vis absorption results. A new resonance Raman band appeared around 190 cm(-1), which was present whenever 5-coordinated Cu-porphyrin existed in Cu(II) Fe(II) hybrid hemoglobins irrespective of the coordination number in Fe(II) subunit. This Raman band might be assigned to Cu-N(epsilon) (His) stretching mode. These results are direct demonstration of the existence of coordination changes of Cu-porphyrin in alpha-subunit within hybrid hemoglobin by shifting the molecular conformation from fully unliganded state to intermediately liganded state. PMID- 11897346 TI - Sequential unfolding of the two-domain protein Pseudomonas stutzeri cytochrome c(4). AB - P. stutzeri cytochrome c(4) is a di-haem protein, composed of two globular domains each with His-Met coordinated haem, and a hydrogen bond network between the domains. The domain foldings are highly symmetric but with specific differences including structural differences of ligand coordination, and different spin states of the oxidised haem groups. We have studied unfolding of oxidised P. stutzeri cyt c(4) induced thermally and by chemical denaturants. Horse heart cyt c was a reference molecule. Isothermal unfolding induced by guanidinium chloride and acid was followed by Soret, alpha/beta, and 701-nm band absorption, and by far-UV circular dichroism spectroscopy. Multifarious patterns emerge, but the two domains clearly unfold sequentially. One phase, assigned to unfolding of the N-terminal domain, proceeds at guanidinium concentrations up to approximately 1.0 M. This is followed by two overlapping phases at higher concentrations. The intermediate state maintains Fe-Met coordination, assigned to the C-terminal domain. Interdomain interaction is reflected in decreasing values of the cooperativity parameters. Differential scanning calorimetry shows a single peak, but two peaks appear when guanidinium chloride up to 0.4 M is present. This reflects different chemical action in chemical and thermal unfolding. Acid induced unfolding kinetics was addressed by pH jumps using diode array stopped flow techniques. Three kinetic phases in the 701 nm Fe-Met marker band, and four phases in the Soret and alpha/beta bands, spanning 4-1000 ms could be distinguished on pH jumps from 7.5 to the range 2.5-3.5. In this range of time and pH cyt c appears to unfold in no more than two phases. Spectral properties of the kinetic intermediates could be identified. Sequential domain unfolding, formation of high-spin states, and an intermediate state with Fe-Met coordination to a single haem group are features of the unfolding kinetics. PMID- 11897347 TI - Electron transfer between cytochrome b(5) and some oxidised haemoglobins: the role of ionic strength. AB - We have compared experimental measurements and Brownian dynamic calculations for the reduction of oxidised adult human haemoglobin by reduced bovine cytochrome b(5) over a range of ionic strengths. Our calculations suggest that the presence of molecular electrostatic fields have a significant role to play in the formation of the electron transfer complexes. These results predict that electron transfer occurs within an ensemble of similarly weakly docked complexes, the formation of which is strongly ionic strength dependent. Application of electron tunneling analysis to the complexes allows us to predict the rates of electron transfer within each ensemble of complexes as a function of ionic strength. The outcome of this theoretical study is compared with experimental rate measurements. A comparison of the results obtained from adult and embryonic haemoglobins, at a fixed ionic strength, indicates a significant difference in the characteristics of complex formation. These data emphasise the role played by electrostatic interactions in this important physiological reaction. PMID- 11897348 TI - Ligand binding reveals protonation events at the active site of cytochrome c oxidase; is the K-pathway used for the transfer of H(+) or OH(-)? AB - We have investigated the CO-recombination kinetics after flash photolysis of CO from the "half-reduced" cytochrome c oxidase as a function of pH. In addition, the reaction was investigated in mutant enzymes in which Lys(I-362) and Ser(I 299), located approximately in the middle of the K-pathway and near the enzyme surface, respectively, were modified. Laser-flash induced dissociation of CO is followed by rapid internal electron transfer from heme a(3) to a. At pH>7 this electron transfer is associated with proton release to the bulk solution (tau congruent with 1 ms at pH 8). Thus, the CO-recombination kinetics reflects protonation events at the catalytic site. In the wild-type enzyme, below pH approximately 7, the main component in the CO-recombination displayed a rate of approximately 20 s(-1). Above pH approximately 7, a slow CO-recombination component developed with a rate that decreased from approximately 8 s(-1) at pH 8 to approximately 1 s(-1) at pH 10. This slow component was not observed with KM(I 362), while with the SD(I-299)/SG(I-299) mutant enzymes at each pH it was slower than with the wild-type enzyme. The results are interpreted in terms of proton release from H(2)O in the catalytic site after CO dissociation, followed by OH(-) binding to the oxidized heme a(3). The CO-recombination kinetics is proposed to be determined by the protonation rate of OH(-) and not dissociation of OH(-), i.e. the K-pathway transfers protons and not OH(-). With the KM(I-362) mutant enzyme the proton is not released, i.e. OH(-) is not formed. With the SD(I 299)/SG(I-299) mutant enzymes the proton is released, but both the release and uptake are slowed by the mutations. During reaction of the reduced enzyme with O(2), the H(2)O at the binuclear center is most likely involved as a proton donor in the O-O cleavage reaction. PMID- 11897349 TI - Isotope effects and intermediates in the reduction of NO by P450(NOR). AB - The mechanism of the heme-thiolate-dependent NADH-NO reductase (P450(NOR)) from Fusarium oxysporum was investigated by kinetic isotope effects including protio, [4S-2H]-, [4R-2H]-, [4,4(2)H(2)]-NADH and stopped-flow measurements. The respective kinetic isotope effects were measured at high NO concentrations and were found to be 1.7, 2.3 and 3.8 indicating a rate-limitation at the reduction step and a moderate stereoselectivity in binding of the cofactor NADH. In a different approach the kinetic isotope effects were determined directly for the reaction of the Fe(III)-NO complex with [4R-2H]- and [4S-2H]-NADH by stopped-flow spectroscopy. The resulting isotope effects were 2.7+/-0.4 for the R-form and 1.1+/-0.1 for the S-form. In addition the 444 nm intermediate could be chemically generated by addition of an ethanolic borohydride solution to the ferric-NO complex at -10 degrees C. In pulse radiolysis experiments a similar absorbing species could be observed when hydroxylamine radicals were generated in the presence of Fe (III) P450(NOR). Based on these results we postulate hydride transfer from NADH to the ferric P450-NO complex resulting in a ferric hydroxylamine-radical or ferryl hydroxylamine-complex and this step, as indicated by the kinetic isotope effects, to be rate-limiting at high concentrations of NO. However, at low concentrations of NO the decay of the 444 nm species becomes the rate-limiting step as envisaged by stopped-flow and optical kinetic measurements in a system in which NO was continuously generated. The last step in the catalytic cycle may proceed by a direct addition of the NO radical to the Fe hydroxylamine complex or by electron transfer from the NO radical to the ferric thiyl moiety in analogy to the postulated mechanisms of prostacyclin and thromboxane biosynthesis by the corresponding P450 enzymes. The latter process of electron transfer could then constitute a common step in all heme-thiolate catalyzed reactions. PMID- 11897350 TI - Pseudomonas aeruginosa cytochrome C(551): probing the role of the hydrophobic patch in electron transfer. AB - Cytochrome c(551) from Pseudomonas aeruginosa is a monomeric redox protein of 82 amino-acid residues, involved in dissimilative denitrification as the physiological electron donor of cd(1) nitrite reductase. The distribution of charged residues on the surface of c(551) is very anisotropic: one side is richer in acidic residues whereas the other shows a ring of positive side chains, mainly lysines, located at the border of an hydrophobic patch which surrounds the heme crevice. In order to map in cytochrome c(551) the surface involved in electron transfer, we have introduced specific mutations in three residues belonging to the hydrophobic patch, namely Val23-->Asp, Pro58-->Ala and Ile59-->Glu. The effect of these mutations was analyzed studying both the self-exchange rate and the electron-transfer activity towards P. aeruginosa cd(1) nitrite reductase, the physiological partner and P. aeruginosa azurin, a copper protein often used as a model redox partner in vitro. Our results show that introduction of a negative charge in the hydrophobic patch severely hampers both homonuclear and heteronuclear electron transfer. PMID- 11897351 TI - A scanning tunneling microscopy (STM) investigation of complex formation between cytochrome P450(cam) and putidaredoxin. AB - We have previously reported the scanning tunnelling microscopy (STM) imaging under buffer of the heme monooxygenase cytochrome P450(cam) from Pseudomonas putida [Faraday Discuss. 116 (2000) 1]. We describe here the adsorption and STM imaging under buffer of complexes of a mutant of cytochrome P450(cam), K344C, and wild-type putidaredoxin (Pdx) on gold(111). The images of Pdx on its own on gold(111) are not uniform, presumably due to multiple orientations of protein adsorption because of the presence of five or more cysteines on the protein surface. STM imaging of a 1:1 mixture of P450(cam)-K344C/Pdx showed a regular array of pairs of different-sized proteins 20-25 A apart arranged in rows across the gold(111) surface which we attribute to the P450(cam)/Pdx complex. The images of the pairs are more regular than those of Pdx on its own, probably as a result of complex formation with P450(cam) partly overcoming the heterogeneity of Pdx adsorption. As far as we are aware this is the first report of STM imaging of a protein/protein complex, and the first direct observation of P450(cam)/Pdx complex formation which is a key step in the catalytic cycle of P450(cam) catalysis. The redox centers of the two proteins are ca. 20 A apart, too far for rapid intracomplex electron transfer. Whether the observed complex is competent for electron transfer or physiologically relevant is not known, and further work is in progress to elucidate the protein-protein interaction. PMID- 11897352 TI - Frontier molecular orbital analysis of Cu(n)-O(2) reactivity. AB - Frontier molecular orbital (FMO) theory coupled with density functional calculations has been applied to investigate the chemical reactivity of three key bioinorganic Cu(n)-O(2) complexes, the mononuclear end-on hydroperoxo-Cu(II), the side-on bridged mu-eta(2):eta(2)-O(2)(2-) Cu(II)(2) dimer and the bis-mu-oxo Cu(III)(2) dimer. Two acceptor orbitals (sigma* and pi*) of each complex and two types of donating substrates (sigma-substrate, phosphine; pi-substrate, alkylbenzene) are considered in the electrophilic attack mechanism. The angular dependences of different reaction pathways are determined using FMO theory and the angular overlap model. Including steric effects, the sigma*/sigma and pi*/pi pathways are found more reactive than the corresponding cross sigma*/pi and pi*/sigma pathways which have poor donor-acceptor orbital overlaps in the sterically constrained substrate access region. PMID- 11897353 TI - An outer-sphere hydrogen-bond network constrains copper coordination in blue proteins. AB - In azurins and other blue copper proteins with relatively low reduction potentials (E(0) [Cu(II)/Cu(I)]<400 mV vs. normal hydrogen electrode), the folded polypeptide framework constrains both copper(II) and copper(I) in such a way as to tune the reduction potentials to values that differ greatly from those for most copper complexes. Largely conserved networks of hydrogen bonds organize and lock the rest of the folded protein structure to a loop that contains three of the ligands to copper. Changes in hydrogen bonds that allow copper(I) to revert more closely to its preferred geometry [relative to the copper(II) geometry] accordingly lead to an increase in E(0). This paper reports mutations in the ligand loop of amicyanin from P. denitrificans that relax the constraints on ligation for copper(I) and significantly raise E(0) for these mutants (for example 415+/-4 mV) relative to that of the native amicyanin (265+/-4 mV). These mutations also shift the pK(a) of a ligand histidine to below 5 relative to 7.0 in the wild type. PMID- 11897354 TI - UV Raman monitoring of histidine protonation and H-(2)H exchange in plastocyanin. AB - UV resonance Raman bands of Cu-bound and protonated histidine residues have been detected in (2)H(2)O solutions of poplar plastocyanin. For the Cu(II) protein, slow NH-(2)H exchange of the His37 ligand was monitored via the growth of bands at 1389 and 1344 cm(-1) when Pcy was exchanged into (2)H(2)O, or via their diminution when the protein was exchanged back into H(2)O; the rate constant is 7 x 10(-4)/s at pH (p(2)H) 7.4 at room temperature. The slow exchange is attributed to imidazole H-bonding to a backbone carbonyl. Nearby bands at 1397 and 1354 cm( 1), appear and disappear within the mixing time, and are assigned to the solvent exposed His87 ligand. The approximately 10 cm(-1) differences between His37 and His87 are attributed to the effect of H-bonding on the imidazole ring modes. The UVRR spectra of the Cu(I) protein in (2)H(2)O reveal a 1408 cm(-1) band, characteristic of NH-(2)H-exchanged histidinium, which grows in as the p(2)H is lowered. Its intensity follows a titration curve with pK(a)=4.6. This protonation is assigned to the His87 residue, whose bond to the Cu(I) is known from crystallography to be broken at low pH. As the 1408 cm(-1) band grows, a band at 1345 cm(-1) diminishes, while another, at 1337 cm(-1) stays constant. These are assigned to modes of bound His87 and His37, respectively, shifted down 7-9 cm(-1) from their Cu(II) positions. PMID- 11897355 TI - Advances in the structure and chemistry of metallothioneins. AB - A low molecular weight (6-7 kDa) class of metalloproteins, designated as metallothioneins (MTs), exhibit repeated sequence motifs of either CxC or CxxC through which mono or divalent d(10) metal ions are bound in polymetallic thiolate clusters. The preservation of metal-thiolate clusters in an increasing number of three-dimensional structures of these proteins signifies the importance of this structural motif. This review focuses on the recent developments regarding the versatile and striking chemical reactivity of MTs as well as on the existence of conformational/configurational dynamics within their structure. Both properties and their interplay are likely to be essential for the still elusive biological function of these proteins. PMID- 11897356 TI - Interaction of anions with rat liver arginase: specific inhibitory effects of fluoride. AB - The inhibitory effects of anions, such as N(3)(-), NO(2)(-), BO(4)(3-), SCN(-), CH(3)COO(-), SO(4)(2-), ClO(4)(-), H(2)PO(4)(-), CN(-), I(-), Br(-), Cl(-) and F( ), on the hydrolysis of L-arginine (L-Arg) by rat liver arginase (RLA) have been studied. From all these anions, only F(-) exhibited a clear inhibitory effect at the mM level. Inhibition of RLA by F(-) is reversible and uncompetitive towards L Arg binding with a K(i) value of 1.3+/-0.5 mM at pH 7.4. This effect is dependent on pH as the IC(50) value of F(-) towards RLA increases from 1.2 to 19 mM when increasing the pH from 7 to 10. Another specific inhibitor of RLA, N(omega) hydroxy-L-nor-arginine (nor-NOHA), that has been recently shown to bind to RLA as a bridging ligand of its (Mn(II))(2) cluster, exhibits some similarities with F( ) in its inhibitory effects (identical pH dependence). It is thus tempting to propose that the inhibitory effects of F(-) could be due to its binding as a bridging ligand of the RLA (Mn(II))(2) cluster. However, further studies are required to determine the modes of interaction of F(-) with RLA. PMID- 11897357 TI - Histidine pK(a) values for the N-lobe of human transferrin: effect of substitution of binding site Asp by Ser (D63S). AB - The pK(a) values have been determined for eight of the nine histidine residues and the amino terminus of the N-lobe of human apo-transferrin (hTF/2N), and for seven of the nine histidine residues and the amino terminus of the protein Asp63Ser hTF/2N containing a mutation of the Fe(3+)-ligand Asp63 to Ser63. Calculations suggested that substitution of aspartate by serine would result in decreases of the pK(a) values of most of the histidine residues in the protein. This was found to be the case experimentally, and allowed assignment of the varepsilonCH resonance of His249. For the wild-type protein, the His residue with a pK(a) of 7.40 was assigned as His249, whereas for the mutant, no observable His residue had a pK(a) value higher than 6.9. The protonated form of His249 appears to be stabilised by interactions with Asp63, and the high pK(a) value may be critical for ensuring the release of iron at endosomal pH (5.5). The mutation lowered the apparent binding constant of hTF/2N for the synergistic anion oxalate from log K 4.0 to log K 3.3. (1)H NMR spectral changes induced by Ga(3+) binding to the mutant are compared to those observed for the wild-type protein. PMID- 11897358 TI - Electron diffraction studies of the calcareous skeletons of bryozoans. AB - Electron microscopy and electron diffraction were used to investigate mineral crystallites dissociated from the skeletal walls of six species belonging to the Bryozoa, a phylum of predominantly marine colony-forming invertebrate animals. Four cheilostome bryozoans (Flustra foliacea, Membranipora membranacea, Thalamoporella novaehollandiae and Cellarinella foveolata) and two cyclostomes (Fasciculipora ramosa and Hornera robusta) were analysed. In each case, an attempt was made to relate the crystal morphology imaged in situ by scanning electron microscopy with the crystallographic orientation of isolated crystals determined by electron diffraction analysis in the transmission electron microscope. The results showed that the calcitic cheilostome and cyclostome skeletons consisted of closely packed arrays of plate-like Mg-containing calcite crystallites, and that the crystallographic a-axis was preferentially aligned perpendicular to the top and bottom surfaces of the flattened particles. The results suggest that calcite biomineralization occurs under similar crystallographic constraints in the five species studied even though the origins of cheilostomes and cyclostomes are separated by over 300 million years in the fossil record of the bryozoans. Similar studies for the aragonite crystallites in skeletons of M. membranacea indicated that the crystallographic b-axis was preferentially oriented perpendicular to the basal surfaces of irregular plate like particles. PMID- 11897359 TI - Assessment of single motor unit conduction velocity during sustained contractions of the tibialis anterior muscle with advanced spike triggered averaging. AB - This paper describes an improved spike triggered averaging technique for the assessment of control properties and conduction velocity (CV) of single motor units (MUs) of the tibialis anterior muscle during voluntary muscle contractions. The method is based on the detection of multi-channel surface EMG signals (with linear electrode arrays) and intramuscularly recorded single MU action potentials (MUAPs). Intramuscular electrodes were inserted in the muscle taking into account the MU structural properties (innervation zone, tendon locations, length of the fibers), assessed by the linear array surface EMG detection technique. An algorithm for intramuscular EMG signal decomposition is used to identify single MUAP trains. The MUAPs detected by the intramuscular EMG decomposition algorithm were used to trigger and average the multi-channel EMG signals. CV of single averaged surface MUAPs was estimated by the use of advanced signal processing methods based on multi-channel recordings which allow to consistently reduce the variance of CV estimates compared with traditional two channel delay estimators. The number of averaged potentials can thus be limited, resulting in high temporal resolution CV estimates. The developed technique was tested on recordings from the tibialis anterior muscle in 11 volunteers during fatigue. It was shown that the method allows the assessment of single MU CV changes (fatigue) as small as 0.1 m/s with less than 2 s data epochs. The method allows reliable assessment of firing rate and conduction properties of single MUs with applications for the investigation of central and peripheral fatigue mechanisms. PMID- 11897360 TI - Voltage-sensitive dye imaging of population neuronal activity in cortical tissue. AB - Voltage-sensitive dyes (VSDs) and optical imaging are useful for studying spatiotemporal patterns of population neuronal activity in cortical tissue. Using a photodiode array and absorption dyes we were able to detect neuronal activity in single trials before it could be detected by local field potential (LFP) recordings. Simultaneous electrical and optical recordings from the same tissue also showed that VSD and LFP signals have different waveforms during different activities, suggesting that they are sensitive to different aspects of the synchronization across the population. Noise, dye bleaching, phototoxicity and optical filter selection are important to the quality of the VSD signal and are discussed in this report. With optimized signal-to-noise ratio (S/N) and total recording time, we can optically monitor approximately 500 locations in an area of 1 mm(2) of cortical tissue with a sensitivity comparable to that of LFP electrodes. The total recording time and S/N of fluorescence and absorption dyes are also compared. At S/N of 8-10, absorption dye NK3630 allows a total recording time of 15-30 min, which can be divided into hundreds of 4-8 s recording trials over several hours, long enough for many kinds of experiments. In conclusion, the VSD method provides a reliable way for examining neuronal activity and pharmacological properties of synapses in brain slices. PMID- 11897361 TI - Signal-to-noise ratio improvement in multiple electrode recording. AB - Recordings of spike trains made with microwires or silicon electrodes include more noise from various sources that contaminate the observed spike shapes compared with recordings using sharp microelectrodes. This is a particularly serious problem if spike shape sorting is required to separate the several trains that might be observed on a particular electrode. However, if recordings are made with an array of such electrodes, there are several mathematical methods to improve the effective signal (spikes) to noise ratio, thus considerably reducing inaccuracy in spike detection and shape sorting. We compare the theoretical basis of three such methods and evaluate their performance with simulated and real data. PMID- 11897362 TI - Artifactual synchrony via capacitance coupling in multi-electrode recording from cat striate cortex. AB - Elucidation of neural connectivity patterns in the brain are thought to give us a mechanistic understanding of how the brain works. Functional connectivity is best studied by simultaneous recording of single-unit activity from many neurons. Accordingly, various types of multiple-microelectrode systems have been developed. We have studied long-range lateral interactions in cat striate cortex. To physiologically characterize interacting cells recorded simultaneously, we used two microelectrodes whose movements were controlled by two independently movable microdrives. The tips of the two microelectrodes were separated by approximately 2 mm or more. During preliminary plotting of two receptive fields of cell pairs, we often noted the emergence of perfectly synchronous firing between two spike trains (amplitude ratio, about 20:1) registered with two microelectrodes. Synchronously firing, smaller spikes disappeared when larger spikes of the pair were lost to either substantial advancement of or placing an electrolytic lesion at the electrode registering the latter. The synchrony also disappeared when two microdrive systems were shielded individually. We concluded that the synchrony was attained through capacitance coupling between two microdrive systems. We proposed a few practical recommendations to avoid the contamination of cross correlograms with the false-positive, narrow peak at time zero due to the presence of reflected spike trains. PMID- 11897363 TI - Automated analysis of global ischemia-induced CA1 neuronal death using terminal UTP nick end labeling (TUNEL). AB - In the brain, DNA fragmentation is associated with apoptotic cell death following ischemic/excitotoxic damage. Fragmented DNA can be detected in situ by labeling the 3'OH termini of the internucleosomal generated fragments with deoxynucleotides, through a process known as terminal deoxynucleotidyl transferase (TdT) dUTP nick end labeling, or TUNEL. TUNEL is frequently being used to assess neuronal death following cerebral ischemia in a number of animal models. However, conventional techniques for TUNEL can be time consuming, and are often subjective and thus can lead to inconsistencies among investigators. Moreover, the lack of tools for its quantification and standardization limits the use of this technique in assessing the magnitude of cell death. In the present report, we describe an improved higher throughput technique for TUNEL staining at room temperature on a BioGenex automated stainer, and its subsequent quantitative analysis using NORTHERN ECLIPSE, an imaging analysis program. Its implementation allows us to effectively quantify TUNEL positive cells in the CA1 region of the hippocampus following global forebrain ischemia in rats. We conclude that this general histological technique can be applied to the study of cell death in numerous other experimental models. PMID- 11897364 TI - Monitoring of idebenone treatment in patients with Friedreich's ataxia by high pressure liquid chromatography with electrochemical detection. AB - Idebenone is a quinone analog that is applied in the treatment of several neurological disorders including Friedreich ataxia and mitochondrial encephalomyopathies. Our aim was to develop an easy and sensitive analytical HPLC procedure for the determination of idebenone in the serum of patients treated with this drug. Serum samples from nine paediatric patients diagnosed with Friedreich ataxia and receiving idebenone treatment were analyzed. Idebenone was separated from serum by reverse high-pressure liquid chromatography and analyzed using an electrochemical detection procedure. No interferences were observed during analysis of patient samples obtained prior to idebenone treatment. Calibration of idebenone concentration indicated a linear range between 500 pmol/l and 5 micromol/l and calculation of within-run and between-run coefficients of variation suggested adequate analytical quality for reliable determination. In agreement with previously reported data, during drug therapy, idebenone serum concentrations (basal conditions, range 0.1-0.49 micromol/l) were greatly elevated 90 min after an oral dose (range 0.66-3.63 micromol/l). Thus, we have developed a simple and rapid method that offers adequate analytical quality for accurate idebenone determination. PMID- 11897365 TI - Analysis of phase-locked oscillations in multi-channel single-unit spike activity with wavelet cross-spectrum. AB - Electrophysiological measures of neural activity frequently display oscillatory patterns at various frequencies. Furthermore, these oscillatory patterns can become dynamically synchronized across a wide region of the brain in a task dependent manner. In this study, phase-locked oscillations in simultaneously recorded spike trains were analyzed using the wavelet cross-spectrum. Adaptation of the existent methods of calculating wavelet cross-spectrum to spike train data was straightforward. In contrast, new methods were needed for evaluating the statistical significance of the cross-spectrum. Although a permutation test based on a large number of re-sampled cross-spectra can provide a reliable estimate of statistical significance, this was quite time-consuming. As an alternative, statistical significance was determined with a normal probability density function estimated from a small number of re-sampled cross-spectra. When applied to neuron pairs recorded in the primate supplementary motor area, the re-sampling procedure produced a reliable outcome even when it was based on as few as ten re sampled cross-spectra. These results suggest that the wavelet analysis in combination with a re-sampling procedure provides a useful tool to examine the dynamic patterns of temporal correlation in cortical spike trains. PMID- 11897366 TI - Bismuth tracing in organotypic cultures of rat hippocampus. AB - Bismuth is known to have neurotoxic side effects in humans and animals. In the 1970s France experienced about a thousand cases of patients suffering from bismuth-induced encephalopathy. Studies suggest that bismuth may provoke a selective degeneration of CA1 pyramidal cells in the organotypic cultures of rat hippocampus. A currently established technique for the histochemical visualization of bismuth was applied on hippocampal tissue cultures allowing the tracing of bismuth in concentrations hitherto not possible. The accumulation and subcellular localization of bismuth is demonstrated in the tissue cultures of rat hippocampus. CA1 pyramidal cells in the rat hippocampus exhibit the highest uptake of bismuth. High bismuth citrate concentrations (10 microM) are able to totally destroy the cytoarchitecture of the hippocampus. At ultrastructural levels bismuth was found to be located exclusively in lysosome-like organelles. PMID- 11897367 TI - Simple behavioral methods to assess the effect of drugs or toxins on sensory experience. AB - When behavioral pharmacologists/toxicologists study conditioned taste aversions (CTAs), or other conditioned responses, as a means to investigate the effects of various drugs or toxins on a learned response, failure to discover a CTA is frequently attributed to the treatment's influence on the associative process. This kind of analysis may fail to identify drug-induced sensory changes that may influence conditioned stimulus (CS) or unconditioned stimulus (US) saliency. The current paper outlines a simple method by which a drug's influence on CS or US sensation may be determined. Further, illustrative data are provided regarding how N-methyl-D-aspartate (NMDA) receptor blockade modulates taste and the sensation of malaise. Ketamine (an NMDA receptor antagonist) has been reported to block CTAs in both neonatal and adult rats. The current experiments evaluated ketamine's ability to modulate the taste of a frequently employed CS (saccharin HCl=SAC) or the aversive aspects of a common US (Lithium Chloride=LiCl). Rats normally exhibit a preference for 0.3% SAC over 0.6% SAC and will suppress consumption of these liquids following an injection of LiCl. We report that ketamine did not markedly antagonize these consummatory patterns nor did it disrupt spontaneous locomotor movements. Taken together, these findings point to ketamine's limited ability to change the sensory capacities required for CTA formation. Investigators interested in determining the underlying causes of drug induced CTA blockade may choose to employ paradigms similar to the one used here. PMID- 11897368 TI - Rapid detection of the prion protein M129V polymorphism with the LightCycler. AB - The common single nucleotide polymorphism at codon 129 of the prion protein gene is a key determinant of the genetic susceptibility to Creutzfeldt-Jakob disease (CJD). Recently, a molecular classification of sporadic CJD based on the M129V genotype in conjunction with other determinants was proposed. In the present study, we describe the development and evaluation of a rapid fluorescent-based assay to detect this polymorphism using the LightCycler system. The two polymorphic alleles could be clearly distinguished by their melting points at 52.1 and 60.4 degrees C, representing the 129V and 129M alleles, respectively. These results were confirmed by DNA sequencing. We evaluated our test in 400 patient samples and found no deviations from the expected melting patterns. The calculated allele frequency for the M-allele was 0.66. Thus, we have established a rapid, reliable fluorescent assay for high-throughput detection of the prion protein M129V polymorphism. PMID- 11897369 TI - The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis. AB - Adult animals continue to produce new neurons in the dentate gyrus of hippocampus. Until now, the principal method of studying neurogenesis has been to inject either tritiated thymidine or 5'-Bromo-2-deoxyuridine (BrdU) intraperitoneally followed by autoradiographic or immunohistochemical detection methods respectively. However, such exogenous markers may produce toxic effects. Our objective was to determine whether Ki-67, a nuclear protein expressed in all phases of the cell cycle except the resting phase, can be used as an alternative, endogenous marker. Using immunohistochemistry, we examined Ki-67 and BrdU expression pattern in rats. Ki-67 was expressed within the proliferative zone of the dentate gyrus and its expression pattern mimicked that of BrdU when examined soon after exogenous BrdU administration. Quantitative comparison of BrdU and Ki 67-positive cells showed 50% higher numbers of the latter when examined 24 h after the BrdU injection. This was expected, since BrdU can be incorporated into DNA only during the S-phase of the mitotic process, whereas Ki-67 is expressed for its whole duration. Experimental increases (by ischemia) or reductions (by radiation) in the number of mitotic cells produced parallel changes in BrdU and Ki-67 signals. Thus, Ki-67 is an effective mitotic marker and has most of the benefits of BrdU and none of the costs. This study provides evidence for Ki-67 to be used as a marker of proliferation in the initial phase of adult neurogenesis. PMID- 11897370 TI - Two variables that can be used as pain indices in experimental animal models of arthritis. AB - Since pain is an important symptom in arthritis, useful behavioral indices for pain in experimental arthritis animal models are important tools for investigative work on arthritis. The purpose of this study was to develop simple and quantifiable behavioral tests, which would represent the level of arthritic pain that develops after induction of inflammation in the knee. Two models of knee joint arthritis were produced: (1) KC model-injection of kaolin and carrageenan into the synovial cavity of the knee, and (2) CFA model-injection of complete Freund's adjuvant into the synovial cavity of the knee. The following three variables were measured before and at various times after the induction of arthritis. As an estimate of the degree of edema, the circumference of the knee was measured. As pain indices, (1) the vocalization threshold of compression force applied to the knee joint was measured to represent tenderness of the joint, and (2) the struggle threshold of the knee extension angle was measured to represent a reduction in range of motion in the arthritic joint. A time course study showed that behavioral changes last for at least 1 week for the KC model and at least 2 weeks for the CFA model. Correlation studies showed that all three variables significantly correlated with each other in both the KC and CFA arthritic models. Systemically injected morphine produced a partial reversal of these indices with the expected time course and dose response of a morphine induced analgesic. It is concluded that two variables, the struggle threshold for knee extension and the vocalization threshold for knee compression, could be used as simple and useful pain indices in experimental models of arthritis. PMID- 11897371 TI - Integrating health law and health policy: a European perspective. AB - Health law is intended to create an environment in which the promotion of health goes hand in hand with the protection of individual rights and the general principles of equality and justice. Over the years, the importance of health law has grown, both at national and international level. As health and human rights are closely interlinked, it is important to integrate health law and health policy. It is to be expected that, especially in Europe, the impact of health law on health policy-making will increase as a result of several developments, e.g. the internationalization of health care and health policy, the issue of consumer protection and the legalization of society. This requires a strategy to stimulate the fruitful relationship between health policy and health law. The most important components of this strategy are discussed. PMID- 11897372 TI - Primary care physicians' attitudes to health care reform in England. AB - Primary care organisation in England has been subject to particularly extensive and far-reaching reform in recent years. In 1991, a quasi-market was introduced into the National Health Service and general practitioners were offered the opportunity to manage independent budgets from which to purchase and deliver care services. Practitioners joined the scheme in increasing numbers, although it was eventually abandoned following a change of government in 1997. This paper reports the results of two surveys of primary care physicians' opinions on the English health care reforms, conducted in 1997 and 2000. It concludes that, first, those opting for discretionary budgets were significantly more supportive of the policy than those not joining the scheme and this support continued long after the scheme had been abolished. Second, professional attitudes, with respect to other terms of service in primary care, remained homogenous to a considerable degree over time. Finally, physicians in favour of imposing user charges tend to be those with responsibility for more patients, suggesting a perceived need to manage patient demand. PMID- 11897373 TI - Reforming China's urban health insurance system. AB - China's urban health insurance system is mainly consisted of labor insurance schemes (LIS) and government employee insurance scheme (GIS). LIS is a work unit based self-insurance system that covers medical costs for the workers and often their dependents as well. GIS covers employees of the State institutions, is financed by general revenues. Since 1980s, China has implemented series of health insurance system reforms, culminating in the government's major policy decision in December of 1998 to establish a social insurance program for urban workers. Compared with the old insurance systems under LIS and GIS, the new system expands coverage to private sector employees and provides a more stable financing with its risk pool at the city level. Despite of these advantages, implementation of China's health insurance reform program is faced with several major challenges, including risk transfer from work units to municipal governments, diverse need and demand for health insurance benefits, incongruent roles of the central and regional governments. These challenges may reflect practical difficulties in policy implementation as well as some deficiencies in the original program design. PMID- 11897374 TI - Health sector reform in South Asia: new challenges and constraints. AB - In early 1990s, Jamison, Mosley and others concluded that a profound demographic and consequent epidemiological transition is taking place in developing countries. According to this classical model, by the year 2015, infectious diseases will account for only about 20% of deaths in developing countries as chronic diseases become more pronounced. These impending demographic and epidemiological transitions were to dominate the health sector reform agenda in developing countries. Following an analysis of fertility, mortality and other demographic and epidemiological data from South Asian and other developing countries, the paper argues that the classical model is in need of re-evaluation. A number of new 'challenges' have complicated the classical interplay of demographic and epidemiological factors. These new challenges include continuing population growth in some countries, rapid unplanned urbanization, the HIV/AIDS pandemic in Sub-Saharan Africa (and its impending threat in South Asia), and globalization and increasing marginalisation of developing countries. While the traditional lack of investment in human development makes the developing countries more vulnerable to the vicissitudes of globalization, increasing economic weakness of their governments forces them to retreat further from the social sector. Pockets of poverty and deprivation, therefore, persist giving rise to three simultaneous burdens for South Asia and much of the rest of the developing world: continuing communicable diseases, increasing burden of chronic diseases, and increasing demand for both primary and tertiary levels of health care services. While these complex factors, on the one hand, underscore the need for health sector reform, on the other, they make the task much more difficult and challenging. The paper emphasizes the need to revisit the classical model of demographic and epidemiological transition. It is argued that the health sector in developing countries must be aware of and effectively address these 'new challenges'. Although it has included data from many developing countries, the focus is primarily on South Asia. PMID- 11897376 TI - Optometrist prescribing of therapeutic agents: findings of the AESOP survey. AB - Throughout the USA and in some parts of Australia and Canada, licensed optometrists may prescribe therapeutic agents for certain eye conditions. However, this role is not currently available to European optometrists. The extension of prescribing rights to new professional groups was the subject of a UK government-commissioned review, which cited optometrists as potential candidates. A recent literature review found limited evidence to assess the appropriateness of eye care delivered by different health care providers. To inform the UK decision, we therefore conducted a national postal survey to explore how optometric practice might change with the introduction of therapeutic prescribing. The Anonymous Enquiry of the Scope for Optometrist Prescribing (AESOP), was sent to a random 10% sample of registered optometrists. Over 80% of respondents indicated that optometrists should be able to train as therapeutic prescribers. Most respondents were willing to undergo training, periodic re accreditation and continuing education, as well to participate in simple professional audit of their prescribing. Respondents anticipated that referrals to general practitioners (GPs) would be reduced by nearly 40% and to ophthalmologists via a GP by nearly 20%. Optometrist participation could increase patient access to therapeutic ocular care by between 29% and 50%. Authorising UK optometrists to prescribe therapeutically for eye diseases would appear to make good use of their existing skills and improve patient access to eye care, while relieving pressures upon other healthcare providers. Tentative economic analysis suggests that the introduction of independent optometrist prescribing may be cost neutral. However, adequate comparative research on the performance of optometrists as prescribers is needed and the issue of reimbursement will require careful consideration. PMID- 11897375 TI - Urgency coding as a dynamic tool in management of waiting lists for psychogeriatric nursing home care in The Netherlands. AB - Criteria are used to prioritise patients on waiting lists for health care services. This is also true for waiting lists for admission to psychogeriatric nursing homes. A patient's position on these latter waiting lists is determined by (changes in) urgency and waiting time. The present article focuses on the process and outcome of an urgency coding system in a fair selection of patients. It discusses the use of urgency codes in the daily practice of waiting list management and the related waiting times. Patients and their informal caregivers were followed from entry on the waiting list to admission to a nursing home. Caregivers were interviewed during the waiting period and after their relative's admission to a nursing home, and the formal urgency codes on the waiting list were monitored. Seventy-eight of the initial 93 patients were admitted to a nursing home. High urgency codes were commonly assigned and the waiting times were shorter for patients with higher urgency codes. Negative consequences of an urgency coding system, e.g. patients with less urgency not being admitted at all and patients not being admitted to the nursing home of their choice, could not be demonstrated. Patients without higher urgency codes were admitted after a mean waiting time of 28 weeks. It may be questioned whether this long waiting time is problematic, because satisfaction of the caregivers with regard to waiting times was not influenced by the actual waiting times. An urgency coding system enables health care professionals to react to changes in the situation of both patients and caregivers by adjusting urgency codes to influence the length of time until nursing home admission. PMID- 11897378 TI - The periviscerokinin (PVK) peptide family in insects: evidence for the inclusion of CAP(2b) as a PVK family member. AB - Periviscerokinins (PVKs) are a distinct insect peptide family with unusual distribution in the central nervous system and neurohemal release sites. PVKs were first isolated from the abdominal perisympathetic organs of Periplaneta americana, but can be found in other insect species. Peptides with structural similarity to PVKs have been identified through searches of the Drosophila genome. The cardioacceleratory peptide CAP(2b) of the hawkmoth Manduca sexta shares close amino acid identity with the PVKs and may thus be included as a structural member of the PVK peptide family. In this review, we provide support for grouping CAP(2b) as a PVK family member based on published sequences, and new immunocytochemical findings and mass spectrometric data. PMID- 11897379 TI - Evidence for the association of FMRFamide-related peptides with the spermatheca of Locusta migratoria. AB - The association of FMRFamide-related peptides (FaRPs) with the spermatheca of Locusta migratoria was demonstrated using radioimmunoassay and immunohistochemical techniques. The physiological effects of various FaRPs on the neurally evoked contractions of the spermatheca were also examined. FMRFamide like immunoreactivity (FLI) was demonstrated in processes and cell bodies situated in the VIIIth (terminal) abdominal ganglion. These included an anterior, central and posterior pair of ventral cell bodies positioned near the midline of the ganglion, in addition to two bilaterally paired dorsal cell bodies in the posterior region of the VIIIth abdominal ganglion. Two axons displaying FLI proceed down the ventral ovipositor nerve (VON) and into the receptaculum seminis nerve which innervates the anterior regions of the spermatheca. FLI was also noted in processes on the spermathecal muscle with the highest density occurring on the spermathecal sac and coil duct. FaRPs applied to the spermathecal muscle included GQERNFLRFamide, NFIRFamide, ADDRNFIRFamide, YGGFMRFamide, FMRFamide, ADVGHVFLRFamide and SchistoFLRFamide (PDVDHVFLRFamide). Dose-dependent physiological effects were only noted for FMRFamide, ADVGHVFLRFamide and SchistoFLRFamide. FMRFamide led to a dose-dependent increase in the amplitude of neurally evoked contractions with a threshold of approximately 5 x 10(-7) M. SchistoFLRFamide, and ADVGHVFLRFamide, had an inhibitory effect, decreasing the amplitude of neurally evoked spermathecal contractions. PMID- 11897380 TI - Identification of the abundant neuropeptide from abdominal perisympathetic organs of locusts. AB - The first member of the periviscerokinin peptide family in Locusta migratoria was identified by post-source decay fragmentation on a MALDI-TOF mass spectrometer using a single neurohemal organ only. The primary sequence of this decapeptide, code-named Lom-PVK, is Ala-Ala-Gly-Leu-Phe-Gln-Phe-Pro-Arg-Val-NH(2). Unlike the situation in cockroaches, Lom-PVK is the only abundant periviscerokinin in L. migratoria. It is present in abdominal perisympathetic organs of various species of locusts and grasshoppers. Its myotropic properties, namely to increase the frequency of the contraction of the heart in L. migratoria and stimulate amplitude and tonus of the locust foregut, is reminiscent of the action of Periplaneta-PVKs. PMID- 11897381 TI - Coherence between biosynthesis and secretion of insect adipokinetic hormones. AB - The importance of the process of continuous biosynthesis of locust adipokinetic hormones (AKHs) for the availability of these peptide hormones for release was assessed in vitro by inhibiting this biosynthesis followed by secretory stimulation. Inhibition of the biosynthetic activity for AKHs by brefeldin A caused a considerable inhibition of the AKH release induced by the endogenous crustacean cardioactive peptide (CCAP). After brefeldin A treatment followed by potassium depolarization, CCAP-induced AKH release was completely abolished. In vitro pulse-chase labeling experiments indicated that constitutive secretion from the AKH-producing cells does not occur. It is concluded that AKH secretion involves a regulated release from a relatively small pool of newly formed secretory granules, while older AKH-containing granules appear to be unavailable for release. PMID- 11897382 TI - New insights in Adipokinetic Hormone (AKH) precursor processing in Locusta migratoria obtained by capillary liquid chromatography-tandem mass spectrometry. AB - After translation, the AKH I and AKH II precursors form three dimeric constructs prior to further processing into the respective AKHs and three dimeric Adipokinetic Hormone Precursor Related Peptides or APRPs (two homodimers and one heterodimer). By capillary liquid chromatography-tandem mass spectrometry we demonstrate that the APRPs in Locusta migratoria are further processed to form two smaller neuropeptides: DAADFADPYSFL (residue 36 to 47 of the AKH I precursor) and YADPNADPMAFL (residue 34 to 45 of the AKH II precursor). The peptides are designated as Adipokinetic Hormone Joining Peptide 1 (AKH-JP I) and 2 (AKH-JP II) respectively. Within the AKH I and AKH II precursor molecules, the classic KK and RR processing sites separate the AKH-JPs from the AKH I and II respectively. At the carboxyterminus, both AKH-JP I and II are flanked by Tyr-Arg, a cleaving site not described before. Such an unusual cleavage site suggests the presence, in the corpora cardiaca, of specific convertases. The AKH-JP-II does not stimulate lipid release from the fat body nor does it stimulate glycogen phosphorylase activity, both key functions of AKH. PMID- 11897383 TI - Cardioacceleratory effects of Manduca sexta allatotropin in the true armyworm moth, Pseudaletia unipuncta. AB - Manduca sexta allatotropin (Manse-AT), a peptide originally isolated on the basis of its ability to stimulate juvenile hormone (JH) biosynthesis in the tobacco hornworm, is a potent in vitro stimulator of the corpora allata (CA) in Pseudaletia unipuncta (Lepidoptera: Noctuidae). At 10(-6)M, Manse-AT stimulated in vitro rates of JH biosynthesis by CA of day 0 and 6 adult females 15- and 10 fold respectively. Both Manse-AT and serotonin were also shown to be dose dependent stimulators of heart rate in day 0, 3 and 6 adult males and females. Furthermore, analysis suggests that there are differences in both resting and Manse-AT-stimulated heart rates depending on age and rearing conditions. PMID- 11897384 TI - Restriction of nutrient intake results in the increase of a specific Manduca sexta allatotropin (Manse-AT) mRNA in the larval nerve cord. AB - The Manduca allatotropin (Manse-AT) gene is expressed as three mRNAs that differ from each other by alternative splicing. The level of one of these mRNAs (RNA-3) is specifically increased in the nerve cord of last instar larvae that were starved, parasitized, or fed the ecdysteroid agonist RH-5992. Each of these treatments results in reduction of feeding and increased levels of juvenile hormone (JH). The normal decline in JH biosynthesis by the corpora allata does not occur in starved or RH-5992-fed larvae. The increase in RNA-3 levels has the capacity to increase the production of Manse-AT and two related peptides that may be part of the complex response of larvae to nutrient deprivation. PMID- 11897385 TI - Effects of allatotropin and allatostatin on in vitro production of juvenile hormones by the corpora allata of virgin females of the moths of Heliothis virescens and Manduca sexta. AB - Retrocerebral complexes (RCs) were isolated from adult females of the moths Heliothis virescens and Manduca sexta. Different homologs of juvenile hormone (JH) produced by the isolated RCs were identified and amounts measured by capillary gas chromatography-chemical ionization (isobutane)-mass spectroscopy. Only JH I, II and III were identified. Incubation of RCs from both species in media containing acetate, but no propionate, induced production of approximately equal amounts of JH II and JH III, but the amount of JH I present was very low in all samples. Incubation of RCs with synthetic Manduca sexta allatotropin stimulated significant increases in production of all three homologs but increases in JH I and JH II were greater than those for JH III. The effect of allatotropin was mimicked by addition of propionate to the medium, which indicated that allatotropin increased supply of acetyl- and propionyl-CoA precursors. Incubation of tissue from H. virescens females during the first 24 h after eclosion with synthetic Manduca sexta allatostatin did not reduce production of JH. However, incubation of tissue from 3-day-old females with allatostatin significantly reduced production of JH. Similarly, incubation of tissue from H. virescens females during the first 24 h after eclosion with both allatotropin and allatostatin did not increase JH over the amount present in extracts from tissue incubated without the neuropeptides, indicating that allatostatin negated the action of allatotropin. Incubation of tissue from H. virescens females with allatostatin plus farnesol or JH III acid resulted in significant production of JH III, but neither JH I nor JH II was detected. These findings indicated that allatostatin acts prior to formation of the sesquiterpene alcohol precursors of JH. PMID- 11897386 TI - The biological activity of diuretic factors in Rhodnius prolixus. AB - The rapid post-feeding diuresis of Rhodnius prolixus is under neurohormonal control and involves the integrated activity of the crop, Malpighian tubules and hindgut. One of the factors which is involved in this rapid diuresis is serotonin, however a peptide(s) is also considered to be involved. In other insects, corticotropin releasing factor (CRF)-like and kinin-like, calcitonin like peptides and CAP(2b) have been demonstrated to be diuretic factors/hormones. In the present study, serotonin and CRF-like peptides increased secretion rate and cAMP content of Rhodnius Malpighian tubules, while the kinin-like peptides tested did not increase secretion rate or cAMP content of the tubules. Extracts of the CNS were processed and several HPLC fractions revealed kinin-like immunoreactivity but these fractions did not increase secretion rate when tested on Malpighian tubules. However, these same fractions did possess activity when tested on the hindgut contraction assay. In addition, material eluting at higher acetonitrile concentrations from the HPLC increased secretion and cAMP content of Rhodnius Malpighian tubules. This material eluted at concentrations of acetonitrile consistent with the elution time of CRF-like peptide standards. Synergism was demonstrated using the pharmacological agent forskolin and serotonin, tested on the rate of secretion of Rhodnius Malpighian tubules, in agreement with data of Maddrell et al. As well, synergism could be demonstrated using mesothoracic ganglionic mass (MTGM) homogenates and serotonin at some concentrations of serotonin. However, combinations of CRF-like material and serotonin increased secretion additively, not synergistically. Kinin-like peptides, tested along with CRF-like material and serotonin, at low concentrations, did not increase secretion above that of those factors tested alone. PMID- 11897387 TI - The effects of crustacean cardioactive peptide on locust oviducts are calcium dependent. AB - The role of calcium as a second messenger in the crustacean cardioactive peptide (CCAP)-induced contractions of the locust oviducts was investigated. Incubation of the oviducts in a calcium-free saline containing, a preferential calcium cation chelator, or an extracellular calcium channel blocker, abolished CCAP induced contractions, indicating that the effects of CCAP on the oviducts are calcium-dependent. In contrast, sodium free saline did not affect CCAP-induced contractions. Co-application of CCAP to the oviducts with preferential L-type voltage-dependent calcium channel blockers reduced CCAP-induced contractions by 32-54%. Two preferential T-type voltage-dependent calcium channel blockers both inhibited CCAP-induced oviduct contractions although affecting different components of the contractions. Amiloride decreased the tonic component of CCAP induced contractions by 40-55% and flunarizine dihydrochloride decreased the frequency of CCAP-induced phasic contractions by as much as 65%, without affecting tonus. Flunarizine dihydrochloride did not alter the proctolin-induced contractions of the oviducts. Results suggest that the actions of CCAP are partially mediated by voltage-dependent calcium channels similar to vertebrate L type and T-type channels. High-potassium saline does not abolish CCAP-induced contractions indicating the presence of receptor-operated calcium channels that mediate the actions of CCAP on the oviducts. The involvement of calcium from intracellular stores in CCAP-induced contractions of the oviducts is likely since, an intracellular calcium antagonist decreased CCAP-induced contractions by 30-35%. PMID- 11897388 TI - Contractions associated with the salivary glands of the blood-feeding bug, Rhodnius prolixus: evidence for both a neural and neurohormonal coordination. AB - The salivary glands of the blood-feeding bug, Rhodnius prolixus, are composed of a single epithelial layer of binucleate cells and a double layer of visceral muscle cells surrounding a large secretory cavity. The saliva contains substances which counteract the hemostasis of the host, and injection of saliva into the host is an essential component of successful and efficient gorging. The muscles surrounding the salivary glands of Rhodnius are under polyneuronal control from the salivary nerve projecting out of the hypocerebral ganglion. The amplitude of contractions induced by neural stimulation is dependent upon both intensity and frequency of nerve stimulation. Serotonin and FMRFamide-related peptides (FaRPs) are delivered in the nerve supply to the salivary glands, and both classes of neuroactive chemicals increase frequency and amplitude of phasic contractions in a dose-dependent manner. A member of the FaRP myosuppressin subfamily, however, inhibits contractions. CRF-related and Leucokinin-like peptides are not delivered in the nerve supply but may be present in the hemolymph during feeding. Leucokinin 1 and Zoone DH (a CRF-related peptide) both induce a dose-dependent increase in basal tonus, with phasic contractions superimposed. Zoone DH is more active than Leucokinin 1. Factors are present in the CNS of Rhodnius which mimic the effects of serotonin and the stimulatory peptides. PMID- 11897389 TI - Diuretic and myotropic activities of N-terminal truncated analogs of Musca domestica kinin neuropeptide. AB - Musca kinin (Musdo-K; NTVVLGKKQRFHSWG-NH(2)) and N-terminal truncated analogs of 4-14 residues in length were assayed for diuretic and myotropic activity on housefly Malpighian tubules and hindgut, respectively. The pentapeptide was the minimum sequence required for biological activity, but it was > 5 orders of magnitude less potent than the intact peptide. The pharmacological profiles of the different analogs in the two assays were very similar, suggesting the same receptor is present on both tissues. Potency was little affected by the deletion of Asn(1), but was reduced > 10-fold after the removal of Thr(2). Deletion of the next 5 residues had relatively little effect, but after the second lysyl residue (Lys(8)) was removed potency fell by one to two orders of magnitude. There was a similar drop in potency after the removal of Arg(10), and at 100 microM the pentapeptide had only 20% of the diuretic activity of the intact peptide. The importance of Arg(10) was confirmed by comparing dose-response curves for Musdo-K [6-15] and Acheta kinin-V (AFSHWG-NH(2)) in the diuretic assay; the substitution of arginine by alanine produced a significant reduction in potency and some loss of activity. PMID- 11897390 TI - cis-peptide bond mimetic tetrazole analogs of the insect kinins identify the active conformation. AB - The insect kinin neuropeptides have been implicated in the regulation of water balance, digestive organ contraction, and energy mobilization in a number of insect species. A previous solution conformation study of an active, restricted conformation cyclic analog, identified two possible turn conformations as the likely active conformation adopted by the insect kinins at the receptor site. These were a cisPro type VI beta-turn over C-terminal pentapeptide core residues 1-4 and a transPro type I-like beta-turn over core residues 2-5, present in a ratio of 60:40. Synthesis and evaluation of the diuretic activity of insect kinin analogs incorporating a tetrazole moiety, which mimics a cis peptide bond, identifies the active conformation as the former. The discovery of a receptor interaction model can lead to the development of potent agonist and antagonist analogs of the insect kinins. Indeed, in this study a tetrazole analog with D stereochemistry has been shown to demonstrate partial antagonism of the diuretic activity of natural insect kinins, providing a lead for more potent and effective antagonists of this critical neuropeptide family. The future development of mimetic agonists and antagonists of insect kinin neuropeptides will provide important tools to neuroendocrinologists studying the mechanisms by which they operate and to researchers developing new, environmentally friendly pest insect control strategies. PMID- 11897391 TI - Metabolism of Manduca sexta allatostatin by hemolymph of larvae of the tomato moth, Lacanobia oleracea. AB - The degradation of synthetic Manduca sexta allatostatin (Manse-AS) by hemolymph from larvae of the tomato moth, Lacanobia oleracea was investigated using reversed phase-high performance liquid chromatography (RP-HPLC), and matrix assisted laser desorption ionisation-time of flight mass spectrometry. Metabolism of 1 nmole Manse-AS in diluted hemolymph was rapid, t(1/2) = 3.5 min, with a number of products produced. Mass spectrometry of HPLC fractions identified cleavage products, which indicated a sequential degradation of Manse-AS from the N-terminal to Manse-AS (7-15). The most abundant products identified were Manse AS (5-15), (6-15), and (7-15). These metabolites were synthesized and assayed for biological activity on juvenile hormone (JH) biosynthesis in vitro. All three of the above deletion peptides showed allatostatin activity, but were not as potent as Manse-AS (1-15). PMID- 11897392 TI - Inactivation of a tachykinin-related peptide: identification of four neuropeptide degrading enzymes in neuronal membranes of insects from four different orders. AB - Tachykinin-related peptides (TRP) are widely distributed in the CNS of insects, where they are likely to function as transmitters/modulators. Metabolic inactivation by membrane ecto-peptidases is one mechanism by which peptide signalling is terminated in the CNS. Using locustatachykinin-1 (LomTK-1, GPSGFYGVRamide) as a substrate and several selective peptidase inhibitors, we have compared the types of membrane associated peptidases present in the CNS of four insects, Locusta migratoria, Leucophaea maderae, Drosophila melanogaster and Lacanobia oleracea. A neprilysin (NEP)-like activity cleaving the G-F peptide bond was the major LomTK-1-degrading peptidase detected in locust brain membranes. NEP activity was also found in Leucophaea brain membranes, but the major peptidase was an angiotensin converting enzyme (ACE), cleaving the G-V peptide bond. Drosophila adult head and larval neuronal membranes cleaved the G-F and G-V peptide bonds. Phosphoramidon inhibited both these cleavages, but with markedly different potencies, indicating the presence in the fly brain of two NEP like enzymes with different substrate and inhibitor specificity. In Drosophila, membrane ACE did not make a significant contribution to the cleavage of the G-V bond. In contrast, ACE was an important membrane peptidase in Lacanobia brain, whereas very little neuronal NEP could be detected. A dipeptidyl peptidase IV (DPP IV) that removed the GP dipeptide from the N-terminus of LomTK-1 was also found in Lacanobia neuronal membranes. This peptidase was a minor contributor to LomTK-1 metabolism by neuronal membranes from all four insect species. In Lacanobia, LomTK-1 was also a substrate for a deamidase that converted LomTK-1 to the free acid form. However, the deamidase was not an integral membrane protein and could be a lysosomal contaminant. It appears that insects from different orders can have different complements of neuropeptide-degrading enzymes. NEP, ACE and the deamidase are likely to be more efficient than the common DPP IV activity at terminating neuropeptide signalling since they cleave close to the C-terminus of the tachykinin, a region essential for maintaining biological activity. PMID- 11897393 TI - Enhanced in vivo activity of peptidase-resistant analogs of the insect kinin neuropeptide family. AB - The diuretic/myotropic insect kinin neuropeptides, which share the common C terminal pentapeptide core FX(1)X(2)WG-NH(2), reveal primary (X(2)-W) and secondary (N-terminal to F) sites of susceptibility to peptidases bound to corn earworm (H. zea) Malpighian tubule tissue. Analogs designed to enhance resistance to tissue-bound peptidases, and pure insect neprilysin and ACE, demonstrate markedly enhanced in vivo activity in a weight gain inhibition assay in H. zea, and strong in vivo diuretic activity in the housefly (M. domestica). The peptidase-resistant insect kinin analog pQK(pQ)FF[Aib]WG-NH(2) demonstrates a longer internal residence time in the housefly than the native muscakinin (MK), and despite a difference of over 4 orders of magnitude in an in vitro Malpighian tubule fluid secretion assay, is equipotent with MK in an in vivo housefly diuretic assay. Aminohexanoic acid (Ahx) is shown to function as a surrogate for N-terminal Lys, while at the same time providing enhanced resistance to aminopeptidase attack. Peptidaese-resistant insect kinin analogs demonstrate enhanced inhibition of weight gain in larvae of the agriculturally destructive corn earworm moth. Potent peptidase resistant analogs of the insect kinins, coupled with an increased understanding of related regulatory factors, offer promise in the development of new, environmentally friendly pest insect control measures. PMID- 11897394 TI - Characterization and baculovirus-directed expression of a myosuppressin encoding cDNA from the true armyworm, Pseudaletia unipuncta. AB - Insect myosuppressins are a highly conserved sub-family of peptides which are primarily characterized by the ability to suppress contraction of visceral muscles in a variety of insect species. We have isolated a cDNA from the true armyworm, Pseudaletia unipuncta, that encodes a prohormone containing a peptide identical to ManducaFLRFamide. We have shown that this myosuppressin gene appears to be expressed in late larval and adult insects. In Manduca sexta, a number of extended-FLRFamide peptides have previously been purified including ManducaFLRFamide, F7D (DPSFLRFamide), F7G (GNSFLRFamide) and two larger peptides F24 and F39 that contain the shorter ManducaFLRFamide sequence at their C terminus. Comparison with the true armyworm prepropeptide characterized here identifies F24 and F39 as partially processed products from the same precursor. Expression in the true armyworm was shown by in situ hybridization to occur in over 150 cells throughout the adult brain and nerve cord, and also to occur in both open and closed endocrine type cells of the gut. Overexpression of the P. unipuncta FLRFamide cDNA from a baculovirus vector in cabbage looper caterpillars was used to assess the potential for myosuppressin expression as a means of enhancing virus efficacy. Viral expression of the armyworm prohormone cDNA resulted in raised levels of RFamide-like products in the hemolymph of infected insects, but the products were found to be chemically distinguishable from authentic mature peptide and probably represent partially processed hormone. PMID- 11897395 TI - Drosophila melanogaster myotropins have unique functions and signaling pathways. AB - Drosophila melanogaster TDVDHVFLRFamide (DMS), SDNFMRFamide, and pEVRFRQCYFNPISCF (FLT) represent three structurally distinct peptide families. Each peptide decreases heart rate albeit with different magnitudes and time-dependent responses. DMS and FLT are expressed in the crop and decrease crop motility; however, SDNFMRFamide expression and effect on the crop has not been reported. These data suggest the peptides have different physiological roles. The peptides have non-overlapping expression patterns in neural tissue, which suggests different mechanisms regulate their synthesis and release. The structures, expression patterns, and activities of the myotropins suggest they have important but different roles in biology and different signaling pathways. PMID- 11897396 TI - Characterization of a putative SchistoFLRFamide receptor in the CNS of Locusta migratoria. AB - A putative SchistoFLRFamide receptor in CNS membrane preparations of Locusta migratoria was characterized by cold competition binding and kinetic binding assays using [125I][Y(1)]SchistoFLRFamide ([125I]YDVDHVFLRFamide) as a radioligand. Binding to this site was saturable, specific, reversible, and of high-affinity. Data fit to a single-site binding model by non-linear regression (r(2) = 0.99) estimated K(d) = 1.73 +/- 0.45 x 10(-9) M and B(max) = 49.0 +/- 12.2 fmol.mg(-1) tissue. Total binding of [125I][Y(1)]SchistoFLRFamide to membrane preparations was reduced in the presence of GTPgammaS, an indication that the putative receptor is G protein-coupled. Structure-activity studies determined that the minimum sequence required for binding was HVFLRFamide. Other aspects of the ligand receptor interaction were also examined. PMID- 11897397 TI - Characterization of a functional neuropeptide F receptor from Drosophila melanogaster. AB - Potential receptors for Drosophila neuropeptide F (DmNPF) were identified in the genome database. One receptor (DmNPFR1) sequence resembled the Lymnaea NPY receptor, an invertebrate homolog of the vertebrate Y-receptor family. DmNPFR1 was cloned and tested for functionality in stably transfected mammalian CHO cells. In whole cell binding assays, DmNPF displaced 125I-NPF in a concentration dependent manner (IC(50) = 65 nM). DmNPF inhibited forskolin-stimulated adenylyl cyclase activity similarly (IC(50) = 51 nM). Whole-mount in situ hybridization revealed that DmNPFR1 RNA is expressed in CNS and midgut of Drosophila larvae. DmNPFR1, a new invertebrate Y-receptor homolog, apparently is a functional receptor for DmNPF. PMID- 11897398 TI - The genomic organization of the open reading frame of the red pigment concentrating hormone gene in the blue crab Callinectes sapidus. AB - The open reading frame (ORF) of the gene for the precursor of the octapeptide Red Pigment Concentrating Hormone (RPCH) from the blue crab Callinectes sapidus was cloned by PCR with oligonucleotides targeted to the initiation and the end of the translation coding sequences. A 272 bp intron was characterized between nucleotides 343 and 344 of the reported cDNA, present in the region coding for the last amino acids of the precursor related peptide of RPCH. The intron genomic structure here described is similar to that reported for the gene coding for the Adipokinetic Hormone (AKH) of the grasshopper Schistocerca nitans. PMID- 11897399 TI - Drosophila melanogaster flatline encodes a myotropin orthologue to Manduca sexta allatostatin. AB - We identified a Drosophila melanogaster gene encoding a peptide that dramatically decreases spontaneous muscle contractions and, correspondingly, named the peptide flatline (FLT). This gene consisted of 4 exons and was cytologically localized to 32D2-3. Processing of a predicted 122 amino acid precursor would release pEVRYRQCYFNPISCF that differs from Manduca sexta allatostatin (Mas-AST) by one amino acid, Y4-->F4. FLT does not act as an allatostatin. In situ tissue hybridization further suggests FLT is a novel brain-gut peptide and specifically, the measured activity indicates that it is a potent myotropin. Despite its profound myotropic effect, pupae injected with FLT eclosed. PMID- 11897400 TI - Synthesis of (S)-3-(1-hydroxy-p-carboran-12-yl)alanine, a novel hydrophobic tyrosine-mimetic for peptides. AB - A new, p -carborane containing analog of tyrosine, 3-[1-hydroxy-1,12-dicarba closo-dodecaboran (12)-12-yl]alanine, was prepared from protected 3-[1-hydroxy 1,12-dicarba-closo-dodecaboran (12)-12-yl]propionic acid in five steps using Oppolzer's sultam methodology for asymmetric hydroxyamination as the key step. The tyrosine mimetic can function as a hydrophobic surrogate for tyrosine residues in insect and mammalian neuropeptides to enhance the lipophilicity, and therefore, the cuticle and/or tissue permeability properties of mimetic analogs. As an amino acid, insertion of the mimic is not limited to the N-terminus but can replace a tyrosine residue at any position within a peptide sequence. PMID- 11897401 TI - A brominated-fluorene insect neuropeptide analog exhibits pyrokinin/PBAN-specific toxicity for adult females of the tobacco budworm moth. AB - An analog of the insect pyrokinin/PBAN class of neuropeptides, which features a 2 amino7-bromofluorene attached to the carboxy-terminal bioactive core of the insect pyrokinin/PBAN class of neuropeptides (Phe-Thr-Pro-Arg-Leu-NH(2)), via a succinnic acid linker, was tested in adult H. virescens moths. This analog was found to induce pheromone production when injected into or applied topically to moths. Topical application of as much as 1 nmol of the analog to moths induced production of significant amounts of pheromone for only 1-2 h, whereas injection of 500 pmol induced pheromone production for up to 20 h. All insects died within 24 h after injection of 500 pmol of the analog. Mortality studies indicated that the LD(50) for the analog was 0.7 pmol when injected. A non-pyrokinin/PBAN peptide analog formed by attachment of 2-amino-7-bromofluorene to Ala-Ala-Arg-Ala Ala-NH(2) (via the succinnic acid linker) did not induce mortality when injected at 1 nmol. Similarly no mortality was found when up to 2 nmol of an analog containing a non-brominated fluorene ring, formed by attachment of 9 fluoreneacetic acid to Phe-Thr-Pro-Arg-Leu-NH(2,) was injected into moths. The data indicated that both the bromine and active core of the pyrokinin neuropeptides (Phe-Thr-Pro-Arg-Leu-NH(2)) were critical for a specific toxic action and suggested that the brominated analog poisoned the moths by interacting with pyrokinin receptors. PMID- 11897402 TI - Insulin-related peptides and their conserved signal transduction pathway. AB - The 'insulin superfamily' is an ancient category of small, structurally related proteins, such as insulin, insulin-like growth factors (IGF) and relaxin. Insulin like signaling molecules have also been described in different invertebrates, including nematodes, mollusks, and insects. They initiate an evolutionary conserved signal transduction mechanism by binding to a heterotetrameric, membrane-spanning receptor tyrosine kinase. Recent physiological and genetic studies have revealed that, in different metazoans, the insulin signaling pathway plays a pivotal role in the regulation of a variety of interrelated, fundamental processes, such as metabolism, growth, reproduction and aging. PMID- 11897403 TI - Neural stem cells and regulation of cell number. AB - Normal CNS development involves the sequential differentiation of multipotent stem cells. Alteration of the numbers of stem cells, their self-renewal ability, or their proliferative capacity will have major effects on the appropriate development of the nervous system. In this review, we discuss different mechanisms that regulate neural stem cell differentiation. Proliferation signals and cell cycle regulators may regulate cell kinetics or total number of cell divisions. Loss of trophic support and cytokine receptor activation may differentially contribute to the induction of cell death at specific stages of development. Signaling from differentiated progeny or asymmetric distribution of specific molecules may alter the self-renewal characteristics of stem cells. We conclude that the final decision of a cell to self-renew, differentiate or remain quiescent is dependent on an integration of multiple signaling pathways and at each instant will depend on cell density, metabolic state, ligand availability, type and levels of receptor expression, and downstream cross-talk between distinct signaling pathways. PMID- 11897404 TI - Comparative analysis of the chemical neuroanatomy of the mammalian trigeminal ganglion and mesencephalic trigeminal nucleus. AB - A characteristic peculiarity of the trigeminal sensory system is the presence of two distinct populations of primary afferent neurons. Most of their cell bodies are located in the trigeminal ganglion (TG) but part of them lie in the mesencephalic trigeminal nucleus (MTN). This review compares the neurochemical content of central versus peripheral trigeminal primary afferent neurons. In the TG, two subpopulations of primary sensory neurons, containing immunoreactive (IR) material, are identified: a number of glutamate (Glu)-, substance P (SP)-, neurokinin A (NKA)-, calcitonin gene-related peptide (CGRP)-, cholecystokinin (CCK)-, somatostatin (SOM)-, vasoactive intestinal polypeptide (VIP)- and galanin (GAL)-IR ganglion cells with small and medium-sized somata, and relatively less numerous larger-sized neuropeptide Y (NPY)- and peptide 19 (PEP 19)-IR trigeminal neurons. In addition, many nitric oxide synthase (NOS)- and parvalbumin (PV)-IR cells of all sizes as well as fewer, mostly large, calbindin D-28k (CB) containing neurons are seen. The majority of the large ganglion cells are surrounded by SP-, CGRP-, SOM-, CCK-, VIP-, NOS- and serotonin (SER)-IR perisomatic networks. In the MTN, the main subpopulation of large-sized neurons display Glu-immunoreactivity. Additionally, numerous large MTN neurons exhibit PV and CB-immunostaining. On the other hand, certain small MTN neurons, most likely interneurons, are found to be GABAergic. Furthermore, NOS-containing neurons can be detected in the caudal and the mesencephalic-pontine junction portions of the nucleus. Conversely, no immunoreactivity to any of the examined neuropeptides is observed in the cell bodies of MTN neurons but these are encircled by peptidergic, catecholaminergic, serotonergic and nitrergic perineuronal arborizations in a basket-like manner. Such a discrepancy in the neurochemical features suggests that the differently fated embryonic migration, synaptogenesis, and peripheral and central target field innervation can possibly affect the individual neurochemical phenotypes of trigeminal primary afferent neurons. PMID- 11897405 TI - Review on the systemic delivery of insulin via the ocular route. AB - Systemic drug absorption from the ocular route is well known. Although there is some absorption from the conjunctival sac, the nasal meatus is the site where the majority of systemic absorption of instilled drug takes place. This article reviews the principles of systemic absorption of insulin applied topically to the eye. The physiological and pharmaceutical considerations for formulation development and the strategy of improving the systemic absorption and bioavailability of insulin are also discussed. PMID- 11897406 TI - Identification and determination of GnRH antagonist gelling at injection site. AB - The purpose of this study was to first observe whether orntide, a GnRH antagonist, gels at the injection site and if so, to develop and validate an extraction method to quantitate the peptide amount as well as assess chemical stability in the gel. After subcutaneous injection of a large dose of orntide acetate solution, a white gel and local traumatized effect were observed at the injection site. Orntide remaining at the injection site was recovered by tissue excision, homogenization and tissue protein precipitation with perchloric acid and quantified by high performance liquid chromatography (HPLC) following separation on a C18 column. The standard curve was linear in the detection range and there was no interference from either blank tissue or excipients of the orntide formulation. The recovery from spiked tissue or that immediately following injection was in the range of 90-110%. MALDI-FT mass spectrometry (MS) of the peak fraction indicated that the orntide recovered from the injection site was in the intact form. The results showed that orntide solution, when injected at a large dose, formed a gel at the injection site. The gel delayed the release from the injection site and caused discernible tissue reaction. PMID- 11897407 TI - 99mTc-glucarate for detection of isoproterenol-induced myocardial infarction in rats. AB - Infarct-avid radiopharmaceuticals are necessary for rapid and timely diagnosis of acute myocardial infarction. The animal model used to produce infarction implies artery ligation but chemical induction can be easily obtained with isoproterenol. A new infarct-avid radiopharmaceutical based on glucaric acid was prepared in the hospital radiopharmacy of the INCMNSZ. 99mTc-glucarate was easy to prepare, stable for 96 h and was used to study its biodistribution in rats with isoproterenol-induced acute myocardial infarction. Histological studies demonstrated that the rats developed an infarct 18 h after isoproterenol administration. The rat biodistribution studies showed a rapid blood clearance via the kidneys. Thirty minutes after 99mTc-glucarate administration the standardised heart uptake value S(h)UV was 4.7 in infarcted rat heart which is six times more than in normal rats. ROIs drawn over the gamma camera images showed a ratio of 4.4. The high image quality suggests that high contrast images can be obtained in humans and the 96 h stability makes it an ideal agent to detect, in patients, early cardiac infarction. PMID- 11897408 TI - Synthesis and evaluation of new oxamniquine derivatives. AB - Oxamniquine derivatives were synthesized and evaluated as novel schistosomicide agents. Oxamniquine (1,2,3,4-tetrahydro-2-[[(1-methylethyl)amino]methyl]-7-nitro 6-quinolinemethanol) was submitted to the Mannich reaction, using formaldehyde, paraformaldehyde and acetaldehyde as reagents, and gave three unexpected products: two of them were cyclized on the alkylamine side chain and another etherified on the aminequinolinemethanol group. The three compounds were biologically evaluated using mice infected with Schistosoma mansoni and showed promising activities, but had higher toxicities. For studies on structure activity relationships, results demonstrate that the side alkylamine group can be modified with preserved activity, but that this modification is associated with increased toxicity. PMID- 11897409 TI - Consistency of Carbopol 971-P NF gels and influence of soluble and cross-linked PVP. AB - A study is made of the polymerization process of polyacrylic acid, commercially known as Carbopol 971 NF, assessing its consistency as a function of the degree of neutralization at pH values from 3 to 12, approximately. Percentage concentrations ranging from 0.1 to 1.4% (w/w) were studied. The gels obtained were non-Newtonian, and pseudoplastic. As concentration and pH rise, the consistency of the gels increase to a maximum, which appears between pH 6 and 8, allowing their use as vehicles in bioadhesive formulations for mucosal application. Over the increasing viscosity interval, functions were obtained to indicate the consistency of the gel as a function of pH and concentration. Since the correlation between the theoretical and experimental results is excellent, the equation found can be used to theoretically calculate the working concentration and pH required to secure the necessary consistency for a given vehicle. The addition of soluble polyvinylpyrrolidone (PVP), and cross-linked PVP (PVPP) does not substantially modify the rheological behavior of the gels, thus permitting their addition to usual vehicles. PMID- 11897410 TI - Characterization of nanoparticle uptake by endothelial cells. AB - Endothelium is an important target for drug or gene therapy because of its important role in the biological system. In this paper, we have characterized nanoparticle uptake by endothelial cells in cell culture. Nanoparticles were formulated using poly DL-lactide-co-glycolide polymer containing bovine serum albumin as a model protein and 6-coumarin as a fluorescent marker. It was observed that the cellular uptake of nanoparticles depends on the time of incubation and the concentration of nanoparticles in the medium. The uptake of nanoparticles was rapid with confocal microscopy demonstrating their localization mostly in the cytoplasm. The mitogenic study demonstrated biocompatability of nanoparticles with the cells. The study thus demonstrates that nanoparticles could be used for localizing therapeutic agents or gene into endothelial cells. Nanoparticles localized in the endothelium could provide prolonged drug effects because of their sustained release characterics, and also could protect the encapsulated agent from enzymatic degradation. PMID- 11897411 TI - Nasal glucagon delivery using microcrystalline cellulose in healthy volunteers. AB - We developed an intranasal powder form of glucagon to improve metabolic status and fatty liver in patients with pancreatectomy. Microcrystalline cellulose, which is commonly used in commercial preparations for allergic rhinitis was used as an absorption enhancer. We compared the intranasal powder form with some spray solutions of glucagon with regard to glucagon absorption, concentration of blood glucose, stability and nasal irritation. The absorption of glucagon from the spray solution including 1.5% sodium glycocholate or 1% sodium caprate was 1.3- and 2.6-fold higher than that from the powder form mixed with microcrystalline cellulose at a ratio of 1:69, respectively. The C(max) values of plasma glucose were 2.18, 3.39 and 1.56 mmol l(-1) in the spray solutions including sodium glycocholate and sodium caprate and in the powder form, respectively. However, glucagon in spray solutions was unstable, but that in the powder form was stable at 5 and 25 degrees C for at least 84 days. The spray solution caused strong irritation, but the powder form did not. These results suggested usefulness of the powder form of glucagon for treatment of pancreatectomized patients. PMID- 11897412 TI - Production of inert cushioning beads: effect of excipients on the physicomechanical properties of freeze-dried beads containing microcrystalline cellulose produced by extrusion-spheronization. AB - Conventional highly compactible fillers such as microcrystalline cellulose (MCC) can be mixed with drug-loaded membrane-coated beads and compressed to form a tablet. However, due to particle size differences, there is substantial risk of segregation leading to weight variation and content uniformity problems. Furthermore, whenever modified release beads are included in a tablet matrix, care must be taken to assure the integrity of the coated beads. This paper describes the development of placebo beads containing MCC whose properties make them uniquely suitable for tableting modified release beads. These placebo beads have high compactibility and the ability to rapidly disintegrate. They deform readily and may provide a high degree of protection to drug-loaded membrane coated beads during compression ('cushioning effect'). They can be produced in size ranges that provide minimal segregation propensity. Beads containing different MCC/lactose ratios and different types and levels of superdisintegrants were produced by extrusion-spheronization followed by freeze drying. The presence of high levels of MCC and different superdisintegrants, especially croscarmellose sodium, increased the granulation liquid requirement, thus producing freeze-dried beads with higher porosities and compactibility. Athy-Heckel analysis studies revealed that beads rich in MCC exhibited lower mean yield pressures than those containing high levels of lactose. The freeze-dried beads exhibited both plastic deformation and brittle fracture characteristics. PMID- 11897413 TI - Transport and metabolism of opioid peptides across BeWo cells, an in vitro model of the placental barrier. AB - In keeping with the advance of biotechnology, cell culture becomes an important tool for investigating the transport and the metabolism phenomena. A cell line of human origin, the BeWo choriocarcinoma cell line, was used for the study of the transport and metabolism of opioid peptides across the in vitro model of the placental barrier. Opioid peptides, both naturally occurring and their synthetic analogs, are of interest to be developed as potent analgesics and were included in this study. The apparent permeability coefficients (Pe)s of the peptides containing 4-11 amino acid or analog residues were in the range of 0.23-14.6 x 10(-5) cm/s. The (Pe)s of these peptides were comparable to those of sucrose or dextrans, hydrophilic markers. The (Pe)s of low molecular weight (MW) peptides was not dependent on their MW or molecular size, whereas an inversely linear correlation between (Pe)s and molecular size was observed with the larger peptides. Molecular sieving of the BeWo monolayer restricted the transport of the peptides with MW> or =1033 Da or molecular size > or =6.6 A. Membrane partitioning ability and charge of the peptides were also investigated and found to be the minor factors regulating the extent of peptide permeation. Contrasting to the transport of Tyr-[D-pen-Gly-Phe-D-Pen] (DPDPE) peptide analog across the blood-brain barrier, the transport of DPDPE across the BeWo monolayers were not indicated to be via carrier-mediated transport. The major transport pathway of the opioid peptides across the BeWo monolayers was found to be via paracellular route. In metabolism studies, aminopeptidase was found to be a major enzyme type responsible for the degradation of naturally occurring peptides but not for the synthetic analogs. The finding obtained from the present study reveals the applicability of the BeWo cell line as an in vitro model for investigating placental transport and metabolism of opioid peptides. PMID- 11897414 TI - Evaluation of the plug formation process of silicified microcrystalline cellulose. AB - To investigate the powder plug formation process of silicified microcrystalline cellulose (SMCC) under compression forces consistent with automatic capsule filling machines, a single-ended saw-tooth wave was used to make powder plugs with different heights (6, 8, 12 mm), at two different punch speeds (1 and 50 mm/s) on a tablet compaction simulator. SMCC was compared to Starch 1500, anhydrous lactose (direct tableting grade), and microcrystalline cellulose. Heckel analysis showed that 'apparent mean yield pressures' (AMYP) of all tested materials increased with an increase in the plug height and punch speed. AMYP appeared to depend on the material type and punch speed. Not all materials fit the Shaxby-Evans relationship at such low compression forces (less than 250 N). Only SMCC 90, SMCC HD90 and anhydrous lactose data fit the equation at both punch speeds. Due to poor axial load transmission, the R values of all tested materials decreased with an increase in the plug height. The experimental data fit the Kawakita equation quite well. Overall, Kawakita's b values were inversely related to AMYP values. The maximum breaking force (MBF) of a 12 mm plug formed at a punch speed of 50 mm/s correlated well with the work of compaction, except for SMCC HD90 and SMCC X, which exhibited very high MBF values. This research demonstrated that several grades of SMCC produced plugs having higher MBF than anhydrous lactose and Starch 1500 under similar compression conditions. The apparently higher compactability of these materials at low plug formation forces may be beneficial in developing direct fill formulations for automatic capsule filling machines. PMID- 11897415 TI - The influence of core materials and film coating on the drug release from coated pellets. AB - The objective of this study was to analyse the influence of the composition of the core of the pellets on the in vitro drug release profile. The different materials (drugs and fillers) were chosen according to their relative solubility. Pellets were prepared by a standardised process of extrusion/spheronisation. A selected fraction size (1-1.4 mm diameter) of pellets of each preparation was coated with Surelease (an aqueous dispersion of ethyl cellulose) to give 5% weight gain. The dissolution studies were performed and data analysed in terms of the Area under the Curve (AUC) of the % dissolved as function of time and Mean Dissolution Time (MDT). ANOVA was applied in order to identify the influence factors and the relationship of cross effects. Canonical analysis and multiple regression were employed to quantify these relationships. The film coat was found to be the major factor controlling the drug release. The results however, show that both drug and filler solubility influenced the drug release profile. Some of the unusual results could only be explained if consideration was given to the physical characteristics of both powder and pellets. In particular, the specific surface area of calcium phosphate compared with other fillers played an important role on the release profile of the model drug. PMID- 11897416 TI - Performance of USP calibrator tablets in flow-through cell apparatus. AB - USP dissolution calibrator tablets were analysed by the flow-through cell method with the intention of examining its applicability for the flow-through cell apparatus suitability test. Test was performed with Dissotest CE-6 apparatus, (Sotax, Switzerland) in flow-through cells for tablets and capsules: smaller cells of 12 mm diameter and larger ones of 22.6 mm diameter. Analyses were performed with laminar and turbulent flow of dissolution medium. The flow rates were 16 and 8 ml/min for laminar flow and only 16 ml/min for turbulent flow. From the results it can be concluded that both salicylic acid tablets and prednisone tablets could be used for apparatus suitability test also for the flow-through cell under the conditions of laminar flows of 16 and 8 ml/min in cells phi 12 and 22.6 mm. As regards the turbulent flow of 16 ml/min, without a holder, cells phi 12 mm could be used for salicylic acid tablets and both cells (phi 12 and 22.6 mm) for prednisone tablets. PMID- 11897417 TI - Liposomes as carriers of amphiphilic gadolinium chelates: the effect of membrane composition on incorporation efficacy and in vitro relaxivity. AB - The effects of membrane composition (phospholipid type and amount of cholesterol), liposome size, drug/lipid ratio (loading) and nature of the amphiphilic gadolinium (Gd) chelate on the incorporation efficacy and magnetic resonance (MR) contrast efficacy (longitudinal (T1) relaxivity) were investigated using a fractional factorial design. A highly lipophilic Gd-chelate was required to ensure complete liposome incorporation. High T1-relaxivity was obtained by using liposomes composed of cholesterol and phospholipids with short acyl chain lengths (dimyristoyl phosphatidyl choline (DMPC) and dimyristoyl phosphatidyl glycerol (DMPG). Two key factors, the loading of Gd-chelate and the amount of cholesterol in small-sized DMPC/DMPG liposomes, were studied further in a central composite optimising design. A robust high relaxivity region was identified, comprising high loading of cholesterol and Gd-chelate. However, the highest T1 relaxivity (52 mM(-1) s(-1)) was found in an area containing no cholesterol and low content of Gd-chelate. Nuclear magnetic resonance dispersion (NMRD) profiles were obtained for five of the liposome compositions from the optimising design, and high relaxivity peaks in the 20 MHz region confirmed the presence of Gd chelates with a long tau(R). A liposome formulation was selected for surface modification with polyethylene glycol (PEG), without having any effect on the T1 relaxivity. PMID- 11897418 TI - Peptidase activity on the surface of the porcine buccal mucosa. AB - Peptide drugs in buccal bioadhesive delivery systems are exposed to the surface of the buccal mucosa at high concentrations over long periods of time. The peptidase activity on the surface of the buccal mucosa has not been evaluated as a barrier to peptide buccal delivery. The in vitro stability of various synthetic substrates on the surface of intact porcine buccal mucosa was determined. No carboxypeptidase or dipeptidyl peptidase IV activity was detected on the buccal mucosa, while aminopeptidase N activity was detected using Leu-p-nitroanilide. No endopeptidase activity was observed towards the peptide substrates. Insulin and insulin B-chain were intact at the 2 h time point at 37 degrees C, while the percent of parent Leu-enkephalin remaining was 18+/-9 (mean+/-S.D., n=9). In the presence of aminopeptidase inhibitors, amastatin, sodium deoxycholate and EDTA, the degradation of Leu-enkephalin was dramatically reduced. This work suggests that the buccal route maybe advantageous for the delivery of peptides that are susceptible to such activities. The inclusion of aminopeptidase inhibitors in buccal bioadhesive delivery systems could improve buccal bioavailability of Leu enkephalin. We suggest that compared with the existing in vitro metabolism methods, the analysis of peptide or protein metabolism on intact buccal mucosa could better predict the degradation of the drug as it crosses the tissue. PMID- 11897419 TI - Development and characterization of solid lipid nanoparticles loaded with magnetite. AB - This paper describes the preparation of colloidal lipid particles containing magnetite from warm emulsions. A two step method was used to obtain the nanoparticles: (i) formulation of a transparent phase by heating a O/W emulsion (aqueous surfactant solution melted with a lipid phase, containing the ethyl oleate and soybean lecithin) in which modified lipophilic magnetite is incorporated, and (ii) preparation of the nanoparticles by dispersing the warm transparent phase in cold water (7 degrees C) under mechanical stirring. The latter method gives spherical nanoparticles of a mean size of 62 nm measured by Photon Correlation Spectroscopy and Transmission Electronic Microscopy. The magnetite entrapment efficiency was determined by use of a magnetophoretic sedimentation method. PMID- 11897420 TI - rhEGF/HP-beta-CD complex in poloxamer gel for ophthalmic delivery. AB - The purpose of the present study is to prepare chemically and physically stable rhEGF/poloxamer gel and to investigate its possibility of ophthalmic delivery. The rhEGF/HP-beta-CD complex markedly increased rhEGF stability compared with rhEGF solution at 4 degrees C. The poloxamer gel was composed of poloxamer 407 (16%) and poloxamer 188 (14%). Additive of rhEGF/HP-beta-CD complexes increased the gelation temperature and 0.5% rhEGF/HP-beta-CD complex exhibited a suitable gelation temperature (35.5 degrees C). The gel strength and bioadhesive force decreased by increasing the rhEGF and HP-beta-CD ratio from 1:4 to 1:20 in the complex. The in vitro release of rhEGF from poloxamer gel containing 1:4 rhEGF/HP beta-CD complex was much slower than that of rhEGF solution and faster than that of 1:20 rhEGF/HP-beta-CD complex. After ocular administration of poloxamer gels in the rabbit, the concentration of rhEGF in tear declined at a first-order elimination. The poloxamer gel containing rhEGF/HP-beta-CD complex increased the area under the concentration-time curve (AUC) of rhEGF in tear fluid compared with gel containing rhEGF solution. 1:20 ratio of rhEGF/HP-beta-CD exhibited high AUC, indicating that rhEGF may be retained in the pre-corneal area for prolonged period. Therefore, the poloxamer gel could be applicable for the development of effective ophthalmic delivery. PMID- 11897421 TI - Assessment of bioequivalence of rifampicin, isoniazid and pyrazinamide in a four drug fixed dose combination with separate formulations at the same dose levels. AB - Tuberculosis (TB) needs treatment with three to five different drugs simultaneously, depending on the patient category. These drugs can be given as single drug preparations or fixed dose combinations (FDCs) of two more drugs in a single formulation. World Health Organization and International Union against Tuberculosis and Lung Disease (IUATLD) recommend FDCs only of proven bioavailability. The relative bioavailability of rifampicin (RIF), isoniazid (INH) and pyrazinamide (PYZ) was assessed on a group of 13 healthy male subjects from a four drug FDC versus separate formulations at the same dose levels. The study was designed to be an open, crossover experiment. A total of nine blood samples each of 3 ml volume were collected over a period of 24-h. The concentrations of RIF, its main metabolite desacetyl RIF (DRIF), INH and PYZ in plasma were assessed by HPLC analysis. Pharmacokinetic parameters namely AUC(0 24), AUC(0-inf), C(max), T(max), were calculated and subjected to different statistical tests (Hauschke analysis, two way ANOVA, normal and log transformed confidence interval) at 90% confidence interval. In addition, elimination rate constant (K(el)) and absorption efficiencies for each drug were also calculated. It was concluded that four drugs FDC tablet is bioequivalent for RIF, INH and PYZ to separate formulation at the same dose levels. PMID- 11897422 TI - Prediction of plasma concentration-time curve of orally administered theophylline based on a scintigraphic monitoring of gastrointestinal transit in human volunteers. AB - The plasma concentration-time profile of theophylline after oral administration in human volunteers was predicted using the individual gastrointestinal (GI) transit data monitored by a gamma scintigraphic technique. Theophylline was administered as aminophylline under fasted and fed condition, along with 99mTc labeled diethylenetriamine-pentaacetic acid (DTPA), an unabsorbable marker to evaluate the GI transit by a gamma scintigraphic technique. Two healthy male volunteers participated under fasted and fed conditions in a crossover study. The GI transit was evaluated by dividing the GI tract to four segments, stomach, jejunum, ileum and cecum/colon. Under the fed condition, the GI transit pattern for each segment was confirmed to alter considerably, causing a delay in the gastric emptying mainly. Further, the plasma concentration curves of theophylline after oral administration were predicted using the GI-Transit-Absorption Model on the basis of individual GI transit parameters calculated by the fitting of the observed data to the GI-Transit Kinetic Model. The absorption rate constant in each segment and the pharmacokinetic parameters after intravenous administration used for the prediction were the values extrapolated from the data in rats and the ones normalized from the values in literatures, respectively. The plasma concentration-time curves for theophylline were well predicted using obtained individual GI transit parameters. The analysis using this method could estimate the variable absorption behavior governed by the GI transit in detail. PMID- 11897423 TI - Design of Peumus boldus tablets by direct compression using a novel dry plant extract. AB - A solid pharmaceutical dosage formulation using a novel dry plant extract of Peumus boldus MOL. (Monimiaceae) (Pb) is proposed. The botanical evaluation of plant material, through morphological and anatomical diagnosis, is presented. This evaluation permits to identify the herb to be used correctly. The analysis of the most extractive solvent mixture and the attainment of plant extract (fluid and dry) are reported. Several formulations (tablets) containing a novel dry plant extract of Pb and common excipients for direct compression are evaluated. The following formulation: dry plant extract of Pb (170 mg), Avicel PH101 (112 mg), Lactose CD (112) and magnesium stearate (6 mg), compressed at 1000 mPa, showed the best pharmaceutical performance. PMID- 11897424 TI - A calorimetric study of phosphocholine membranes mixed with desmopressin and its diacylated prodrug derivative (DPP). AB - The influence of the water-soluble peptide, desmopressin (DDAVP) and its dipalmitoylated prodrug derivative (DPP) on the thermal behaviour of three different saturated phosphatidylcholine lipid membranes was investigated by differential scanning calorimetry. For lipid membranes composed of dimyristoyl, dipalmitoyl and distearoyl phosphatidylcholines the addition of DDAVP at concentrations of up to 10 mol% resulted in an insignificant change in the thermodynamic phase behaviour. In contrast, the dipalmitoylated DPP prodrug caused major changes on the lipid membrane phase behaviour manifested as a drastic decrease in the heat capacity peak height and a concomitant broadening of the main phase transition as well as a decrease in the transition enthalpy. In addition, the main phase transition temperature was slightly decreased and the pre-transition of the three phosphatidylcholines was abolished when DPP was present. PMID- 11897425 TI - The effect of ionizing radiation on some derivatives of 1,4-dihydropyridine in the solid state. AB - The effect of gamma and beta radiation in doses between 10 and 100 kGy on physico chemical properties of four derivatives of 1,4-dihydropyridine (nifedipine, nitrendipine, felodipine and nimodipine) in the solid state was analysed. A number of qualitative and quantitative methods such as UV, IR, TLC, GLC, DSC, EPR as well as organoleptic and gravimetric analysis were used to determine and analyse any changes resulting from irradiation. In order to determine the effectiveness of sterilization with ionizing radiation of doses from 10 to 25 kGy, various microbiological tests were used. It was established that only doses 10-20 kGy of both kinds of radiation ensure total sterilization without any degradation of physico-chemical properties of the compounds studied. For the doses 50-100 kGy a decrease in the content of the compounds, appearance of the products of their decomposition and changes in the melting point and IR spectra appeared. Felodipine (with chlorophenyl substituent) was found to be much more sensitive to ionising radiation than nifedipine, nitrendipine and nimodipine (all with nitrophenyl substituent). PMID- 11897427 TI - The effect of process variables on the degradation and physical properties of spray dried insulin intended for inhalation. AB - The aim of this study was to investigate the effect of process variables on the degradation and physical properties of spray dried insulin intended for inhalation. A 2(4) full factorial experimentally designed study was performed to investigate the influence of the following independent spray drying variables: feed flow rate, nozzle gas flow rate, inlet air temperature and aspirator capacity (drying gas flow rate). Human insulin (biosynthetic and Ph.Eur. quality) was dissolved in distilled water to concentrations of 5 mg/ml. The solutions were spray dried in a Mini Spray Dryer Buchi and the dry powders produced were characterized by high performance liquid chromatography, size exclusion chromatography, laser diffraction, thermo gravimetric analysis, scanning electron microscopy and weighing. The degradation of insulin was found to be affected mainly by the process variables that determine the outlet air temperature, i.e.: inlet air temperature, aspirator capacity and feed flow rate. The outlet air temperature should be kept below 120 degrees C to avoid degradation. A statistical optimization of the spray drying variables was performed, and found to recommend an experiment with an outlet air temperature of 61+/-4 degrees C. This experiment ought to generate a yield of 54+/-7% by weight of particles with a mass median diameter 2.9+/-0.4 microm, moisture content 3.9+/-0.5% by weight, content of high molecular weight proteins 0.3+/-0.1% by area, A-21 desamido insulin 0.3+/-0.05% by area and other insulin related compounds 0.3+/-0.1% by area. PMID- 11897426 TI - Controlled drug release properties of ionically cross-linked chitosan beads: the influence of anion structure. AB - By adopting a novel chitosan cross-linked method, i.e. chitosan/gelatin droplet coagulated at low temperature and then cross-linked by anions (sulfate, citrate and tripolyphosphate (TPP)), the chitosan beads were prepared. Scanning electron microscopy (SEM) observation showed that sulfate/chitosan and citrate/chitosan beads usually had a spherical shape, smooth surface morphology and integral inside structure. Cross-sectional analysis indicated that the cross-linking process of sulfate and citrate to chitosan was much faster than that of TPP due to their smaller molecular size. But, once completely cross-linked, TPP/chitosan beads possessed much better mechanical strength and the force to break the beads was approximately ten times higher than that of sulfate/chitosan or citrate/chitosan beads. Release media pH and ionic strength seriously influenced the controlled drug release properties of the beads, which related to the strength of electrostatic interaction between anions and chitosan. Sulfate and citrate cross-linked chitosan beads swelled and even dissociated in simulated gastric fluid (SGF) and hence, model drug (riboflavin) released completely in 5 h; while in simulated intestinal fluid (SIF), beads remained in a shrinkage state and drug released slowly (release % usually <70% in 24 h). However, swelling and drug release of TPP/chitosan bead was usually insensitive to media pH. Chitosan beads, cross-linked by a combination of TPP and citrate (or sulfate) together, not only had a good shape, but also improved pH-responsive drug release properties. Salt weakened the interaction of citrate, especially sulfate with chitosan and accelerated beads swelling and hence drug release rate, but it was insensitive to that of TPP/chitosan. These results indicate that ionically cross linked chitosan beads may be useful in stomach specific drug delivery. PMID- 11897428 TI - Preparation and characterization of sterile and freeze-dried sub-200 nm nanoparticles. AB - The feasibility of producing sterile and freeze-dried polyester nanoparticles was investigated. Various poly(D,L-lactide-co-glycolide) and poly(D,L-lactide) were selected as biodegradable polymers. Using the salting-out procedure, process parameters were optimized to obtain sub-200 nm particles. After purification, the nanoparticle suspensions containing different lyoprotectants were sterilized by filtration. Freeze-drying was performed using vials covered with 0.22 microm membrane filters in order to preserve the suspensions from bacterial contamination. Sterility was assessed on the final product according to pharmacopoeial requirements using the membrane filtration method. With all polymers tested, sub-200 nm particles could be obtained. Nanoparticles with a size as low as 102 nm were prepared with good reproducibility and narrow size distribution. Upon freeze-drying, it appeared that complete redispersion of all types of polyester nanoparticles could be obtained in presence of the lyoprotectants tested such as saccharides while aggregation was observed without lyoprotectant. Sterility testing showed no microbial contamination indicating that sterile nanoparticulate formulations have been achieved. PMID- 11897429 TI - Carbapenem antibiotics inhibit valproic acid transport in Caco-2 cell monolayers. AB - The concomitant use of carbapenem antibiotics with valproic acid has been prohibited because carbapenems induced a decrease in plasma concentration of valproic acid in epileptic patients during valproic acid therapy. Our previous in vivo study in rats proposed that inhibition by carbapenem of the intestinal absorption of valproic acid might be a possible mechanism for the drug-drug interaction. To demonstrate the hypothesis, we examined the effects of imipenem and panipenem on intestinal transepithelial transport of valproic acid using Caco 2 cell monolayers. Imipenem and panipenem inhibited the transport of [14C] valproic acid across the Caco-2 cell monolayers from apical-to-basolateral side in a concentration-dependent manner, although they had no effect on the uptake of [14C]-valproic acid by Caco-2 cells. The inhibition by the carbapenems of the valproic acid transport was found even when they were added to only the basolateral side. From these results, the carbapenems may inhibit the absorption of valproic acid at the basolateral membrane of intestinal epithelial cells, which contributes to the decrease in plasma concentration of valproic acid after oral administration. PMID- 11897430 TI - Simultaneous optimization of percutaneous delivery and adhesion for ketoprofen poultice. AB - Topical poultices of ketoprofen were prepared using deionized water, propylene glycol (X1), and glycerin (X2) as the vehicle in combination with hydrophilic matrix materials, including gelatin (X3) and sodium polyacrylate. A mixture design was utilized to evaluate the influence of these constituents (X1-X3) on the adhesion of the poultice and the percutaneous penetration of ketoprofen from poultices. The adhesion of the poultice was measured based on the L-Peel test method using a Tensile and Compression Testing Machine. Percutaneous delivery was conducted using nude mouse skin as the barrier. The poultice containing the highest weight fraction of gelatin demonstrated the highest value of peak stress, whereas the poultice containing 0% weight fraction of gelatin showed the smallest value among all formulations. This indicates that gelatin was the main factor determining the adhesion of the poultice. However, the interactive influence of propylene glycol with gelatin on the adhesion of the poultice cannot be ignored. On the contrary, the formulation having the maximal penetration rate was determined to be the vehicle with 0% weight fraction of gelatin and the highest percent weight fraction of glycerin. This indicates that the presence of glycerin in the poultice was able to increase the flux of ketoprofen to some extent. Quantification of individual's effect based on this mixture design resulted in a polynomial equation: Peak stress=0.033X1+0.016X2+0.12X3, flux=1.90X1+4.70X2 6.65X3. Finally, an optimized formulation with acceptable adhesion and a flux comparable to two commercial products was developed in this study. PMID- 11897431 TI - Refractory angina pectoris: mechanism and therapeutic options. AB - As the survival of patients with primary coronary events continues to increase, the number of patients presenting with coronary artery disease unsuitable to further revascularization techniques and symptoms refractory to medical therapy also continues to rise. The aims of this review were to define the population of patients with refractory angina pectoris and to present the therapeutic options currently available for this condition. Refractory angina pectoris is defined, and traditional medical therapies are discussed. Then, current therapeutic options for patients with refractory angina are extensively reviewed. A multitude of therapeutic options exist for patients with refractory angina pectoris. Small, uncontrolled studies have shown a potential benefit for additional antiplatelet and antithrombotic therapy. In randomized trials, neurostimulation has been shown to be effective in reducing angina symptoms. Enhanced external counterpulsation is a viable treatment option for select patients with refractory angina. In many randomized trials, laser revascularization has been shown to diminish angina symptoms, although no placebo-controlled studies exist to date. Gene therapy is a promising area of research in this field. Percutaneous in situ coronary venous arterialization is in its infancy, but may be able to treat many patients if proved successful. No data support the role of chelation therapy in this population. Heart transplantation remains a final option for these patients. Further research of the techniques mentioned in this review is warranted. The importance of randomized, double-blinded, placebo-controlled trials cannot be overemphasized, as the placebo effect of these therapies is probably marked. PMID- 11897432 TI - Particulate air pollution induces progression of atherosclerosis. AB - OBJECTIVES: We sought to determine the effect of exposure to air pollution particulate matter <10 microm (PM(10)) on the progression of atherosclerosis in rabbits. BACKGROUND: Epidemiologic studies have associated exposure to ambient PM(10) with increased cardiovascular morbidity and mortality. We have previously shown that PM(10) exposure induces a systemic inflammatory response that includes marrow stimulation, and we hypothesized that this response accelerates atherosclerosis. METHODS: Watanabe heritable hyperlipidemic rabbits were exposed to PM(10) (n = 10) or vehicle (n = 6) for four weeks, and bone marrow stimulation was measured. Quantitative histologic methods were used to determine the morphologic features of the atherosclerotic lesions. RESULTS: Exposure to PM(10) caused an increase in circulating polymorphonuclear leukocytes (PMN) band cell counts (day 15: 24.6 +/- 3.0 vs. 11.5 +/- 2.7 x 10(7)/l [PM(10) vs. vehicle], p < 0.01) and an increase in the size of the bone marrow mitotic pool of PMNs. Exposure to PM(10) also caused progression of atherosclerotic lesions toward a more advanced phenotype. The volume fraction (vol/vol) of the coronary atherosclerotic lesions was increased by PM(10) exposure (33.3 +/- 4.6% vs. 19.5 +/- 3.1% [PM(10) vs. vehicle], p < 0.05). The vol/vol of atherosclerotic lesions correlated with the number of alveolar macrophages that phagocytosed PM(10) (coronary arteries: r = 0.53, p < 0.05; aorta: r = 0.51, p < 0.05). Exposure to PM(10) also caused an increase in plaque cell turnover and extracellular lipid pools in coronary and aortic lesions, as well as in the total amount of lipids in aortic lesions. CONCLUSIONS: Progression of atherosclerosis and increased vulnerability to plaque rupture may underlie the relationship between particulate air pollution and excess cardiovascular death. PMID- 11897433 TI - Air pollution as a cause of heart disease. Time for action. PMID- 11897434 TI - Clinical benefits of low serum digoxin concentrations in heart failure. AB - OBJECTIVES: We sought to determine whether there was a relationship between serum digoxin concentration (SDC), including SDCs typically regarded as low, and clinical efficacy related to digoxin in patients with symptomatic left ventricular dysfunction. BACKGROUND: Digitalis glycosides have been used for 200 years in the treatment of heart failure (HF), but the SDC required for optimal clinical efficacy and acceptable toxicity remains controversial. METHODS: This relationship was investigated by utilizing data from two randomized, double blinded, placebo-controlled, digoxin-withdrawal trials: the Prospective Randomized study Of Ventricular failure and Efficacy of Digoxin (PROVED) and the Randomized Assessment of Digoxin on Inhibitors of Angiotensin-Converting Enzyme (RADIANCE). Major end points were worsening HF, change in left ventricular ejection fraction and treadmill time after randomization. The primary analysis investigated the relationship between SDC at randomization and these end points. A secondary categorical analysis compared these end points in patients who discontinued digoxin versus patients who continued digoxin and had low (0.5 to 0.9 ng/ml), moderate (0.9 to 1.2 ng/ml) or high (>1.2 ng/ml) SDCs at randomization. RESULTS: Multiple regression analysis failed to find a relationship between randomization SDC, considered as a continuous variable, and any study end point (all p > 0.236). Multivariable Cox analysis found that the risk of worsening HF was significantly less (all p < 0.02) for patients in any category of SDC who continued digoxin, as compared with patients withdrawn from digoxin. Specifically, patients in the low SDC category were significantly less likely than placebo patients to experience worsening HF during follow-up (p = 0.018). CONCLUSIONS: The beneficial effects of digoxin on common clinical end points in patients with HF were similar, regardless of SDC. PMID- 11897435 TI - Low-dose digoxin in patients with heart failure. Less toxic and at least as effective? PMID- 11897436 TI - Plasma oxidized low-density lipoprotein as a prognostic predictor in patients with chronic congestive heart failure. AB - OBJECTIVES: The aim of this study was to evaluate the relationship between plasma oxidized low-density lipoprotein (oxLDL), a marker of oxidative stress, and the prognosis of patients with chronic congestive heart failure (CHF). BACKGROUND: Oxidative stress appears to play a role in the pathophysiology of CHF. We have recently reported the usefulness of plasma oxLDL as a marker of oxidative stress in CHF patients with dilated cardiomyopathy. METHODS: We measured the plasma level of oxLDL by sandwich enzyme-linked immunosorbent assay using a specific monoclonal antibody against oxLDL in 18 age-matched normal subjects and in 84 patients with chronic CHF (New York Heart Association functional class II to IV) and monitored them prospectively for a mean follow-up period of 780 days. RESULTS: Plasma oxLDL level was significantly higher in severe CHF patients than in control subjects and mild CHF patients. A significant negative correlation existed between the plasma level of oxLDL and left ventricular ejection fraction (LVEF) and a significant positive correlation between the plasma level of oxLDL and plasma norepinephrine level. Twenty-six patients had cardiac events; 14 had cardiac death and 12 were hospitalized for heart failure or other cardiovascular events. Among 10 variables including LVEF and neurohumoral factors, only high plasma levels of brain natriuretic peptide and oxLDL were shown to be independent predictors of mortality. CONCLUSIONS: These results indicate that the plasma level of oxLDL is a useful predictor of mortality in patients with CHF, suggesting that oxidative stress plays an important role in the pathophysiology of CHF. PMID- 11897437 TI - Left ventricular assist device in end-stage heart failure: persistence of structural myocyte damage after unloading. An immunohistochemical analysis of the contractile myofilaments. AB - OBJECTIVES: We sought to evaluate the contractile proteins in cardiomyocytes of patients with end-stage heart failure (HF) before and after mechanical support with a left ventricular assist device (LVAD). BACKGROUND: Improvement of myocyte dysfunction has been suggested after LVAD support. METHODS: Fourteen patients' myocardial biopsies taken at the time of LVAD implantation and after explantation, at the time of heart transplantation, were processed for routine hematoxylin-eosin staining and immunohistochemistry using monoclonal antibodies against actin, myosin, tropomyosin, troponin C and T and titin. A grading scale from 1 (abnormal staining of all myocytes, no cross-striation) to 5 (normal fiber anatomy and striation) was used. The cross-sectional area of cardiomyocytes was also measured. RESULTS: The cardiomyocytes' cross-sectional area decreased after support, from 519 +/- 94 microm(2) to 319 +/- 53 microm(2) (p < 0.001). Actin, tropomyosin, troponin C, troponin T and titin at the time of LVAD implantation showed widespread distortion of architecture; their grades were 1.4 +/- 0.6, 2.3 +/- 1.0, 2.1 +/- 0.9, 2.1 +/- 1.2 and 2.0 +/- 0.6, respectively. In contrast, myosin morphology was preserved (4.6 +/- 0.7). After LVAD support, actin, tropomyosin, troponin C, troponin T and titin showed improvement (grades 2.7 +/- 1.3 [p = 0.004], 3.2 +/- 1.2 [p = 0.021], 3.3 +/- 0.9 [p = 0.004], 3.0 +/- 1.1 [p = 0.048] and 3.1 +/- 0.9 [p = 0.001], respectively), but no normalization. The myosin pattern deteriorated slightly (3.6 +/- 1.6 [p = 0.058]). CONCLUSIONS: After LVAD support, during a period of 213 +/- 135 days in patients with end stage HF, despite a decrease in the size of the cardiomyocytes, severe structural myocyte damage persisted. This does not support complete recovery of myocyte histologic features. PMID- 11897439 TI - Are we clear about the mechanisms by which biopsy evidence of interstitial fibrosis following cardiac transplantation helps predict late post-transplant coronary artery disease? PMID- 11897438 TI - Myocardial ischemic-fibrotic injury after human heart transplantation is associated with increased progression of vasculopathy, decreased cellular rejection and poor long-term outcome. AB - OBJECTIVES: We sought to assess the influence of peritransplant ischemia and fibrosis on the development of allograft vasculopathy, acute cellular rejection and long-term outcome. BACKGROUND: Allograft vasculopathy is a common long-term complication of cardiac transplantation. One of the potential risk factors is peritransplant allograft ischemia. METHODS: One hundred forty heart transplant recipients had baseline and one-year intravascular ultrasound analysis done to assess the progression of allograft vasculopathy. Serial endomyocardial biopsies were evaluated for cellular rejection, vascular rejection, ischemia and fibrosis. Based on histology, patients were classified into one of the following groups: nonischemic (n = 32), ischemia (n = 24), fibrosis (n = 62) or vascular rejection (n = 22). Three-color flow cytometry crossmatching (FCXM) was used to assess donor-specific human lymphocyte antigens (HLA) sensitization. Long-term outcome of patients in each group was assessed by estimating incidence of graft failure or deaths over a seven-year follow up. RESULTS: Patients in the fibrosis group had the lowest incidence of donor-specific HLA sensitization (40%, p = 0.008) and lowest average episodes of cellular rejection (1.7 +/- 1.4, p = 0.04), but they had increased coronary vasculopathy progression (change in coronary intimal thickness = 0.59 +/- 0.28 mm, p < 0.0001) and poor seven-year event-free survival (49%, p = 0.01). CONCLUSIONS: The development of fibrosis after cardiac transplantation is associated with advanced coronary vasculopathy, although a low incidence of acute cellular rejection is noted, suggesting the presence of nonimmune mechanisms in mediating the pathogenesis of allograft vasculopathy. PMID- 11897440 TI - Autosomal dominant dilated cardiomyopathy with atrioventricular block: a lamin A/C defect-related disease. AB - OBJECTIVES: We investigated the prevalence of lamin A/C (LMNA) gene defects in familial and sporadic dilated cardiomyopathies (DCM) associated with atrioventricular block (AVB) or increased serum creatine-phosphokinase (sCPK), and the corresponding changes in myocardial and protein expression. BACKGROUND: It has been reported that familial DCM, associated with conduction disturbances or variable myopathies, is causally linked to LMNA gene defects. METHODS: The LMNA gene and myocardial ultrastructural and immunochemical changes were analyzed in 73 cases of DCM (49 pure, 15 with AVB [seven familial, eight sporadic], 9 with increased sCPK), four cases of familial AVB and 19 non-DCM heart diseases. The normal controls included eight heart donor biopsies for tissue studies and 107 subjects for LMNA gene studies. RESULTS: Five novel LMNA mutations (K97E, E111X, R190W, E317K, four base pair insertion at 1,713 cDNA) were identified in five cases of familial autosomal dominant DCM with AVB (5/15: 33%). The LMNA expression of the myocyte nuclei was reduced or absent. Western blot protein analyses of three hearts with different mutations showed an additional 30-kDa band, suggesting a degrading effect of mutated on wild-type protein. Focal disruptions, bleb formation and nuclear pore clustering were documented by electron microscopy of the myocyte nuclear membranes. None of these changes and no mutations were found in the nine patients with DCM and increased sCPK or in the disease and normal controls. CONCLUSIONS: The LMNA gene mutations account for 33% of the DCMs with AVB, all familial autosomal dominant. Increased sCPK in patients with DCM without AVB is not a useful predictor of LMNA mutation. PMID- 11897441 TI - Reversible regional wall motion abnormalities on exercise technetium-99m-gated cardiac single photon emission computed tomography predict high-grade angiographic stenoses. AB - OBJECTIVES: We sought to determine the level of angiographic stenosis at which reversible regional wall motion abnormalities (RWMA) are present on exercise stress technetium-99m (Tc-99m)- gated single photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI), and whether assessments of stress and rest RWMA add incremental diagnostic information. BACKGROUND: Stress and rest gated SPECT MPI enables the detection of post-exercise stunning. Although some studies have correlated RWMA to the severity of MPI defects, only one previous study correlated RWMA on gated MPI to angiographic findings. However, this correlation excluded patients with rest perfusion defects and did not involve gating of rest images. METHODS: One hundred patients undergoing angiography within six months of exercise stress Tc-99m (sestamibi)-gated SPECT MPI (in the absence of interim cardiac events or revascularization) were recruited. Images were acquired 15 to 30 min after stress and interpreted without knowledge of the Duke treadmill score, left ventricular ejection fraction and angiographic data. RESULTS: The sensitivity of reversible RWMA for angiographic stenoses >70% was 53%, with a specificity of 100%. The presence of reversible RWMA was able to stratify patients with angiographic stenoses of 50% to 79% and 80% to 99% with a high positive predictive value. A good correlation was noted between the presence of reversible RWMA and the coronary artery jeopardy score (R = 0.49, p < 0.0001). Multivariate analysis showed that the post-stress RWMA, Duke treadmill and reversible RWMA scores were significant predictors of angiographic severity. CONCLUSIONS: Post-stress and reversible RWMA, as shown by exercise stress Tc-99m gated SPECT MPI, are significant predictors of angiographic disease and add incremental value to MPI for the assessment of angiographic severity. PMID- 11897442 TI - Low event rate for stress-only perfusion imaging in patients evaluated for chest pain. AB - OBJECTIVES: We sought to demonstrate the safety of stress-only perfusion imaging among patients with low to medium probability of coronary disease being evaluated for chest pain. BACKGROUND: The need for performing rest in addition to stress perfusion imaging to confirm normalcy is due largely to defects created on the stress images by attenuation artifacts. A low cardiac event rate among patients undergoing stress-only imaging with attenuation correction (AC) would validate the safety of stress-only imaging. METHODS: Patients with low to medium pretest probability for coronary artery disease (CAD) referred for chest pain evaluation from July 1, 1997 to July 1, 1999, were scheduled for a two-day stress/rest tomographic (single photon emission computerized tomography, or SPECT) perfusion imaging study with Tc-99m sestamibi. Patients were imaged on a variable angle camera with AC using Gd-153 scanning line sources (Vantage ADAC, Milpitas, California). If the stress scan was normal without AC or corrected with AC, the patient did not return for rest scan and was followed. RESULTS: Seven hundred twenty-nine patients underwent stress-only imaging, and follow-up was obtained on 652 (89%) of those patients. There were 224 males and 428 females with mean age of 52 +/- 13 years. Mean follow-up was 22.3 +/- 6.4 months. The mean pretest probability was 37 +/- 24%. The non-AC images showed breast and/or diaphragmatic attenuation artifacts severe enough to have required the patient to return for rest imaging in 37% of patients, and all corrected completely with AC. During follow-up, there were two noncardiac deaths and no cardiac deaths. There was one myocardial infarction; three patients with progressive unstable angina underwent diagnostic coronary angiography showing significant CAD. The overall cardiac event rate was 0.6%. CONCLUSIONS: These results support stress-only imaging in patients with low to medium probability for CAD as a safe, time- and cost efficient imaging modality. PMID- 11897443 TI - Increased central pulse pressure and augmentation index in subjects with hypercholesterolemia. AB - OBJECTIVES: The goal of this study was to investigate the relation between serum cholesterol, arterial stiffness and central blood pressure. BACKGROUND: Arterial stiffness and pulse pressure are important determinants of cardiovascular risk. However, the effect of hypercholesterolemia on arterial stiffness is controversial, and central pulse pressure has not been previously investigated. METHODS: Pressure waveforms were recorded from the radial artery in 68 subjects with hypercholesterolemia and 68 controls, and corresponding central waveforms were generated using pulse wave analysis. Central pressure, augmentation index (AIx) (a measure of systemic stiffness) and aortic pulse wave velocity were determined. RESULTS: There was no significant difference in peripheral blood pressure between the two groups, but central pulse pressure was significantly higher in the group with hypercholesterolemia (37 +/- 11 mm Hg vs. 33 +/- 10 mm Hg [means +/- SD]; p = 0.028). Augmentation index was also significantly higher in the patients with hypercholesterolemia group (24.8 +/- 11.3% vs. 15.6 +/- 12.1%; p < 0.001), as was the estimated aortic pulse wave velocity. In a multiple regression model, age, short stature, peripheral mean arterial pressure, smoking and low-density lipoprotein cholesterol correlated positively with AIx, and there was an inverse correlation with heart rate and male gender. CONCLUSIONS: Patients with hypercholesterolemia have a higher central pulse pressure and stiffer blood vessels than matched controls, despite similar peripheral blood pressures. These hemodynamic changes may contribute to the increased risk of cardiovascular disease associated with hypercholesterolemia, and assessment may improve risk stratification. PMID- 11897444 TI - Pulse transmission coefficient: a novel nonhyperemic parameter for assessing the physiological significance of coronary artery stenoses. AB - OBJECTIVES: We sought to test the hypothesis that the pulse transmission coefficient (PTC) can serve as a nonhyperemic physiologic marker for the severity of coronary artery stenosis in humans. BACKGROUND: Coronary lesions may impair the transmission of pressure waves across a stenosis, potentially acting as a low pass filter. The PTC is a novel nonhyperemic parameter that calculates the transmission of high-frequency components of the pressure signal through a stenosis. Thus, it may reflect the severity of the coronary artery stenosis. This study was designed to examine the correlation between PTC and fractional flow reserve (FFR) in patients with coronary artery disease. METHODS: Pressure signals were obtained by pressure guidewire in 56 lesions (49 patients) in the nonhyperemic state and were analyzed with a new algorithm that identifies the high-frequency components in the pressure signal. The PTC was calculated as the ratio between the distal and proximal high-frequency components of the pressure waveform across the lesion. The FFR measurements were assessed with intracoronary adenosine. RESULTS: There was a significant correlation between PTC and FFR (r = 0.81, p < 0.001). By using a receiver operating characteristic analysis, we identified a PTC < 0.60 (sensitivity 100%, specificity 98%) to be the optimal cutoff value for predicting an FFR < 0.75. CONCLUSIONS: Pulse transmission coefficient is a novel nonhyperemic parameter for the physiologic assessment of coronary artery stenoses. It correlates significantly with FFR and may predict an FFR < 0.75 with high accuracy. Pulse transmission coefficient may be useful as an adjunct measurement to FFR, especially in patients with microcirculatory disease and impaired maximal hyperemia. PMID- 11897446 TI - Management of coronary artery fistulae. Patient selection and results of transcatheter closure. AB - OBJECTIVES: We report short-term findings in 33 patients after transcatheter closure (TCC) of coronary artery fistulae (CAF) and compare our results with those reported in the recent transcatheter and surgical literature. BACKGROUND: Transcatheter closure of CAF has been advocated as a minimally invasive alternative to surgery. METHODS: We reviewed all patients presenting with significant CAF between January 1988 and August 2000. Those with additional complex cardiac disease requiring surgical management were excluded. RESULTS: Of 39 patients considered for TCC, occlusion devices were placed in 33 patients (85%) at 35 procedures and included coils in 28, umbrella devices in 6 and a Grifka vascular occlusion device in 1. Post-deployment angiograms demonstrated complete occlusion in 19, trace in 11, or small residual flow in 5. Follow-up echocardiograms (median, 2.8 years) in 27 patients showed no flow in 22 or small residual flow in 5. Of the 6 patients without follow-up imaging, immediate post deployment angiograms showed complete occlusion in 5 or small residual flow in 1. Thus, complete occlusion was accomplished in 27 patients (82%). Early complications included transient ST-T wave changes in 5, transient arrhythmias in 4 and single instances of distal coronary artery spasm, fistula dissection and unretrieved coil embolization. There were no deaths or long-term morbidity. Device placement was not attempted in 6 patients (15%), because of multiple fistula drainage sites in 4, extreme vessel tortuosity in 1 and an intracardiac hemangioma in 1. CONCLUSIONS: A comparison of our results with those in the recent transcatheter and surgical literature shows similar early effectiveness, morbidity and mortality. From data available, TCC of CAF is an acceptable alternative to surgery in most patients. PMID- 11897445 TI - Intensive cholesterol reduction lowers blood pressure and large artery stiffness in isolated systolic hypertension. AB - OBJECTIVES: We sought to investigate the effects of intensive cholesterol reduction on large artery stiffness and blood pressure in normolipidemic patients with isolated systolic hypertension (ISH). BACKGROUND: Isolated systolic hypertension is associated with elevated cardiovascular morbidity and mortality and is primarily due to large artery stiffening, which has been independently related to cardiovascular mortality. Cholesterol-lowering therapy has been efficacious in reducing arterial stiffness in patients with hypercholesterolemia, and thus may be beneficial in ISH. METHODS: In a randomized, double-blinded, cross-over study design, 22 patients with stage I ISH received three months of atorvastatin therapy (80 mg/day) and three months of placebo treatment. Systemic arterial compliance was measured noninvasively using carotid applanation tonometry and Doppler velocimetry of the ascending aorta. RESULTS: Atorvastatin treatment reduced total and low-density lipoprotein cholesterol and triglyceride levels by 36 +/- 2% (p < 0.001), 48 +/- 3% (p < 0.001) and 23 +/- 5% (p = 0.003), respectively, and increased high density lipoprotein cholesterol by 7 +/- 3% (p = 0.03). Systemic arterial compliance was higher after treatment (placebo vs. atorvastatin: 0.36 +/- 0.03 vs. 0.43 +/- 0.05 ml/mm Hg, p = 0.03). Brachial systolic blood pressure was lower after atorvastatin treatment (154 +/- 3 vs. 148 +/- 2 mm Hg, p = 0.03), as were mean (111 +/- 2 vs. 107 +/- 2 mm Hg, p = 0.04) and diastolic blood pressures (83 +/- 1 vs. 81 +/- 2 mm Hg, p = 0.04). There was a trend toward a reduction in pulse pressure (71 +/- 3 vs. 67 +/- 2 mm Hg, p = 0.08). CONCLUSIONS: Intensive cholesterol reduction may be beneficial in the treatment of patients with ISH and normal lipid levels, through a reduction in large artery stiffness. PMID- 11897447 TI - Intrinsic sinus and atrioventricular node electrophysiologic adaptations in endurance athletes. AB - OBJECTIVES: In the present study, we evaluated sinus and atrioventricular (AV) node electrophysiology of endurance athletes and untrained individuals before and after autonomic pharmacologic blockade. BACKGROUND: Endurance athletes present a higher prevalence of sinus bradycardia and AV conduction abnormalities, as compared with untrained individuals. Previous data from our laboratory suggest that nonautonomic factors may be responsible for the longer AV node refractory period found in well-trained athletes. METHODS: Six aerobically trained male athletes and six healthy male individuals with similar ages and normal rest electrocardiograms were studied. Maximal oxygen uptake (O(2)max) was measured by cardiopulmonary testing. The sinus cycle length (SCL), AV conduction intervals, sinus node recovery time (SNRT), Wenckebach cycle (WC) and anterograde effective refractory period (ERP) of the AV node were evaluated by invasive electrophysiologic studies at baseline, after intravenous atropine (0.04 mg/kg) and after addition of intravenous propranolol (0.2 mg/kg). RESULTS: Athletes had a significantly higher O(2)max as compared with untrained individuals. The SCL was longer in athletes at baseline, after atropine and after the addition of propranolol for double-autonomic blockade. The mean maximal SNRT/SCL was longer in athletes after atropine and after propranolol. The WC and anterograde ERP of the AV node were longer in athletes at baseline, after atropine and after propranolol. CONCLUSIONS: Under double-pharmacologic blockade, we demonstrated that sinus automaticity and AV node conduction changes of endurance athletes are related to intrinsic physiology and not to autonomic influences. PMID- 11897448 TI - Cerebral blood flow velocity declines before arterial pressure in patients with orthostatic vasovagal presyncope. AB - OBJECTIVES: We studied hemodynamic changes leading to orthostatic vasovagal presyncope to determine whether changes of cerebral artery blood flow velocity precede or follow reductions of arterial pressure. BACKGROUND: Some evidence suggests that disordered cerebral autoregulation contributes to the occurrence of orthostatic vasovagal syncope. We studied cerebral hemodynamics with transcranial Doppler recordings, and we closely examined the temporal sequence of changes of cerebral artery blood flow velocity and systemic arterial pressure in 15 patients who did or did not faint during passive 70 degrees head-up tilt. METHODS: We recorded photoplethysmographic arterial pressure, RR intervals (electrocardiogram) and middle cerebral artery blood flow velocities (mean, total, mean/RR interval; Gosling's pulsatility index; and cerebrovascular resistance [mean cerebral velocity/mean arterial pressure, MAP]). RESULTS: Eight men developed presyncope, and six men and one woman did not. Presyncopal patients reported light-headedness, diaphoresis, or a sensation of fatigue 155 s (range: 25 to 414 s) before any cerebral or systemic hemodynamic change. Average cerebral blood flow velocity (CBFV) changes (defined by an iterative linear regression algorithm) began 67 s (range: 9 to 198 s) before reductions of MAP. Cerebral and systemic hemodynamic measurements remained constant in nonsyncopal patients. CONCLUSIONS: Presyncopal symptoms and CBFV changes precede arterial pressure reductions in patients with orthostatic vasovagal syncope. Therefore, changes of cerebrovascular regulation may contribute to the occurrence of vasovagal reactions. PMID- 11897449 TI - Three-dimensional nonfluoroscopic mapping and ablation of inappropriate sinus tachycardia. Procedural strategies and long-term outcome. AB - OBJECTIVES: We conducted this study to assess long-term results of three dimensional (3-D) mapping-guided radiofrequency ablation (RFA) of inappropriate sinus tachycardia (IST). Change in activation after the administration of esmolol was also assessed and compared to the shift documented with successful sinus node (SN) modification. BACKGROUND: The long-term results after RFA of IST have been reported to vary between 27% and 66%. METHODS: Thirty-nine patients (35 women, mean age 31 +/- 9 years) with debilitating IST were included in the study. The area around the earliest site of activation recorded using the 3-D mapping system was targeted for ablation. The shift in the earliest activation site after administration of esmolol was compared with the shift after RFA. RESULTS: The heart rate at rest and in drug-free state ranged between 95 and 125 beats/min (mean 99 +/- 14 beats/min). Sinus node was successfully modified in all patients. Following ablation, the mean heart rate dropped to 72 +/- 8 beats/min, p < 0.01. The extent of the 3-D shift in caudal activation along the crista terminalis was more pronounced after RFA than during esmolol administration (23 +/- 11 mm vs. 7 +/- 5 mm, respectively, p < 0.05). No patient required pacemaker implantation after a mean follow-up time of 32 +/- 9 months; 21% of patients experienced recurrence of IST and were successfully re-ablated. CONCLUSIONS: Three dimensional electroanatomical mapping seems to facilitate and improve the ablation results of IST. The difference in caudal shift seen after esmolol administration and following SN modification suggests that adrenergic hypersensitivity is not the only mechanism responsible for the inappropriate behavior of the SN. PMID- 11897450 TI - Gender differences and normal left ventricular anatomy in an adult population free of hypertension. A cardiovascular magnetic resonance study of the Framingham Heart Study Offspring cohort. AB - OBJECTIVES: We sought to derive gender-specific cardiovascular magnetic resonance (CMR) reference values for normative left ventricular (LV) anatomy and function in a healthy adult population of clinically relevant age. BACKGROUND: Cardiovascular magnetic resonance imaging is increasingly applied in the clinical setting, but age-relevant, gender-specific normative values are currently unavailable. METHODS: A representative sample of 318 Framingham Heart Study (FHS) Offspring participants free of clinically overt cardiovascular disease underwent CMR examination to determine LV end-diastolic and end-systolic volume (EDV and ESV, respectively), mass, ejection fraction (EF) and linear dimensions (wall thickness, cavity length). Subjects with a clinical history of hypertension or those with a systolic blood pressure > or =140 mm Hg or diastolic pressure > or =90 mm Hg at any FHS cycle examination were excluded, leaving 142 subjects (63 men, 79 women; age 57 +/- 9 years). RESULTS: All volumetric (EDV, ESV, mass) and unidimensional measures were significantly greater (p < 0.001) in men than in women and remained greater (p < 0.02) after adjustment for subject height. Volumetric measures were greater (p < 0.001) in men than in women after adjustment for body surface area (BSA), but there were increased linear dimensions in women after adjustment for BSA. In particular, end-diastolic dimension indexed to BSA was greater in women (p < 0.001) than in men. There were no gender differences in global LVEF (men = 0.69; women = 0.70). CONCLUSIONS: Cardiovascular magnetic resonance measures of LV volumes, mass and linear dimensions differ significantly according to gender and body size. This study provides gender-specific normal CMR reference values, uniquely derived from a population-based sample of persons free of cardiovascular disease and clinical hypertension. These data may serve as a reference to identify LV pathology in the adult population. PMID- 11897451 TI - Early and late complications associated with transcatheter occlusion of secundum atrial septal defect. AB - OBJECTIVES: The goal of this study was to report the early and late complications experienced in atrial septal defect (ASD) transcatheter closure. BACKGROUND: Atrial septal defect transcatheter occlusion techniques have become an alternative to surgical procedures. A number of different devices are available for transcatheter ASD closure. The type and rate of complications are different for different devices. METHODS: Between December 1996 and January 2001, 417 patients (mean age: 26.6 +/- 19 years) underwent transcatheter occlusion of secundum type ASD. Complications were categorized into major and minor. Two different devices were used: the CardioSEAL/STARFlex in 159 patients and the Amplatzer septal occluder in 258 patients. RESULTS: Thirty-four patients experienced 36 complications during the hospitalization (8.6%, 95% confidence interval: 6.1% to 11.1%). Ten patients underwent elective surgical repair because of device malposition (three patients) or device embolization (seven patients). Twenty-four patients experienced 25 minor complications: unsatisfactory device position or embolization. Devices were retrieved using a gooseneck snare and/or a basket; 11 patients experienced arrhythmic problems. Other complications were: pericardial effusion, thrombus formation on the left atrial disc, right iliac vein dissection, groin hematoma, hemorrhage in the retropharynx and sizing balloon rupture. Two patients had late complications: peripheral embolization in the left leg one year after implantation of an Amplatzer device and sudden death 1.5 year later. CONCLUSIONS: Our series of patients with ASD by transcatheter occlusion shows that the procedure is safe and effective in the vast majority of cases. To further reduce the complications rate, the criteria of device selection according to ASD morphology and some technical tips during implantation are discussed. PMID- 11897452 TI - Long-term outcome of patients with ventricular septal defect considered not to require surgical closure during childhood. AB - OBJECTIVES: The purpose of the study was to assess the long-term outcome of patients with small ventricular septal defects (VSDs) considered not to require surgical closure during childhood. BACKGROUND: Although patients with small VSDs have generally been considered not to require surgery, more recent data suggest that a significant percentage of these patients develop serious problems during adult life. METHODS: A total of 229 consecutive patients (115 females) with a VSD considered too small to require surgery during childhood as defined by normal pulmonary artery pressure, less than 50% shunt, pulmonary vascular resistance < or =200 dynes x s cm(-5), no VSD-related aortic regurgitation (AR), and no symptoms and who had no additional hemodynamically relevant heart defect were followed in an adult congenital heart disease program. Physical examination, electrocardiography, and echocardiography were performed in all patients in one- to three-year intervals; exercise tests and Holter monitoring were performed in 140 and 127 patients, respectively. RESULTS: Follow-up was completed in 222 patients (97%). Mean age at last visit was 30 +/- 10 years. Spontaneous VSD closure was observed in 14 patients (6%). No patients died, four patients (1.8%) had an episode of endocarditis, of whom two required aortic valve replacement, and one additional patient (0.4%) had surgical closure for hemodynamic reasons. For 118 patients who entered the study between 1993 and 1996 and were prospectively followed for 7.4 +/- 1.2 years, event-free survival with end points defined as death, endocarditis or heart surgery was 99.1 +/- 0.8% at three years, 96.5 +/- 1.7% at six years and 95.5 +/- 1.9% at eight years. At last visit, 94.6% of all patients studied were symptom free. Left ventricular (LV) size by echocardiography was normal in 198 (89%) patients, borderline in 23 patients and definitely enlarged in only one patient. None had systolic LV dysfunction, and pulmonary artery pressure (PAP) was normal in all patients. Mean exercise capacity was 92 +/- 21% of expected, and 87% of patients had no arrhythmias on Holter monitoring, with the remainder showing benign rhythm disorders. CONCLUSIONS: Outcome in well-selected patients with a small VSD is good. Surgical closure does not appear to be required during childhood as long as left-to-right shunt is <50% and signs of LV volume overload are absent, when PAP is not elevated, and no VSD-related AR or symptoms are present. PMID- 11897453 TI - Overproduction of platelet microparticles in cyanotic congenital heart disease with polycythemia. AB - OBJECTIVES: We sought to clarify the role of platelets in the pathogenesis of abnormal coagulation in patients with cyanotic congenital heart disease (CCHD) with polycythemia; we evaluated the production of platelet microparticles (MPs), platelet degranulation and aggregation response, as well as the correlations of these variables with polycythemia. BACKGROUND: A shortened life span and suppressed aggregability of platelets are well known in patients with CCHD. Although platelet MPs are overproduced and play an important role in the coagulation process in various hematologic and cardiovascular disorders, the production of MPs remains to be elucidated in CCHD. We studied 19 patients who had CCHD with polycythemia and 21 age-matched subjects with acyanotic congenital heart disease (ACHD). Flow cytometry, using monoclonal antibodies, showed the presence of MPs as particles positive for the surface antigen (glycoprotein IIb/IIIa) specific to platelets, and platelet alpha-degranulation was recognized as platelets positive for the surface antigen of P-selectin. Platelet aggregation was assessed as the response to adenosine diphosphate (ADP). Relationships between these indexes and hematocrit (Hct) values were also evaluated. RESULTS: Production of MPs correlated positively with Hct and markedly increased at Hct values above 60% in patients with CCHD. Surface P-selectin and the mean platelet volume in patients with CCHD were comparable with those in patients with ACHD. The platelet aggregation response to ADP significantly and negatively correlated with Hct. In two subjects who showed hemoptysis and underwent phlebotomy, MPs were reduced 6 h after the procedure. CONCLUSIONS: Platelet MPs are overproduced in patients who have CCHD with polycythemia, probably due to a high shear stress derived from blood hyperviscosity. Circulating incompetent platelets, which have already been activated, as well as MPs, might play an important role in the coagulation abnormalities identified in such patients. PMID- 11897456 TI - Viewing tobacco use in movies: does it shape attitudes that mediate adolescent smoking? AB - BACKGROUND: Social cognitive theory posits that children develop intentions and positive expectations (utilities) about smoking prior to initiation. These attitudes and values result, in part, from observing others modeling the behavior. This study examines, for the first time, the association between viewing tobacco use in movies and attitudes toward smoking among children who have never smoked a cigarette. DESIGN/SETTING: Cross-sectional school-based survey was used among randomly selected Vermont and New Hampshire middle schools. The sample consisted of 3766 middle school students (grades 5-8). The sample was primarily white and equally distributed by gender. The primary exposure was number of movie tobacco-use occurrences viewed. We first counted occurrences of tobacco use in each of 601 recent popular motion pictures. Each student was asked to select movies they had seen from a random subset of 50 movies. Based on movies the adolescent had seen, movie tobacco-use occurrences were summed to determine exposure . The outcome was susceptibility to smoking, positive expectations, and perceptions of smoking as normative behavior for adolescents or adults. RESULTS: The movies in this sample contained a median of five occurrences of tobacco use (interquartile range=1, 12). The typical adolescent never-smoker had viewed 15 of the 50 movies on his/her list. From movies adolescents reported seeing, exposure to movie tobacco-use occurrences varied widely: median=80, and interquartile range 44 to 136. The prevalence of susceptibility to smoking increased with higher categories of exposure: 16% among students who viewed 0 to 50 movie tobacco occurrences; 21% (51 to 100); 28% (101 to 150); and 36% (>150). The association remained statistically significant after controlling for gender, grade in school, school performance, school, friend, sibling and parent smoking, sensation-seeking, rebelliousness, and self-esteem. Compared with adolescents exposed to < or =50 occurrences of tobacco use, the adjusted odds ratio of susceptibility to smoking for each higher category was 1.2 (95% confidence interval 0.9, 1.5), 1.4 (1.1, 1.9), and 1.6 (1.3, 2.1), respectively. Similarly, higher exposure to tobacco use in movies significantly increased the number of positive expectations endorsed by the adolescent and the perception that most adults smoke, but not the perception that most peers smoke. CONCLUSIONS: This study provides empirical evidence that viewing movie depictions of tobacco use is associated with higher receptivity to smoking prior to trying the behavior. PMID- 11897457 TI - Effect of the incident at Columbine on students' violence- and suicide-related behaviors. AB - BACKGROUND: This study examined the impact that the violent incident at Columbine High School may have had on reports of behaviors related to violence and suicide among U.S. high school students. METHODS: Nationally representative data from the 1999 Youth Risk Behavior Survey (YRBS) were analyzed using logistic regression analyses. RESULTS: Students who completed the 1999 YRBS after the Columbine incident were more likely to report feeling too unsafe to go to school and less likely to report considering or planning suicide than were students who completed the 1999 YRBS before the incident. CONCLUSIONS: These results highlight how an extreme incident of school violence can affect students nationwide. PMID- 11897458 TI - The impact of drug pricing policies on the health of the elderly. AB - BACKGROUND: Prices for prescription drugs vary widely in the United States, from the prices charged by retail pharmacies to 50% below average wholesale prices (AWP), depending on the purchaser's bargaining power. At higher drug prices, expenditures for essential preventive interventions, such as those aimed at hypertension and hypercholesterolemia, buy less health for the elderly. The magnitude of this effect, however, is unknown. METHODS: Cost-effectiveness analyses of drug interventions, which estimate their health effects and costs, sometimes include analyses of the impact of drug prices. We use results from studies of preventive drugs used by the elderly to calculate the life-years that could be purchased for $1 million of expenditure at the AWP, and at prices 20% and 40% below the AWP. This range reflects the range of prices in the United States today. RESULTS: Drug prices have a substantial effect on the amount of health that can be purchased for $1 million, especially among elderly people with several health conditions. For example, at 40% below the AWP, $1 million spent on statins yields 90 years of life for patients aged 75 to 84 with a history of myocardial infarction. At the AWP, the number of life-years for $1 million drops to 48--a loss of 42 life-years. CONCLUSIONS: A Medicare drug benefit program that supports prices at the high end of the current range could yield substantially less health for the elderly, and one that promotes differential pricing could promote unequal access to preventive medications--and thus to health--by Medicare beneficiaries across the country. PMID- 11897460 TI - Missed opportunities to immunize: psychosocial and practice correlates. AB - BACKGROUND: The goal of this pilot study was to correlate missed opportunities to immunize young children with providers' psychosocial characteristics and self reported immunization practices. METHODS: In a population of children aged 0 to 36 months, missed opportunities to immunize were established for a sample of 28 providers, who also responded to a valid and reliable instrument measuring the aforementioned variables. RESULTS: Missed opportunities were significantly lower among providers with higher vested interest (r=-0.45, p=0.02) and tended to be lower among providers with more positive attitudes toward having all children properly immunized at every healthcare visit (r=-0.33, p =0.09). Neither knowledge nor perceived barriers correlated significantly with missed opportunities. Providers missed opportunities to immunize in over half of the visits studied (mean, 0.58), yet all of them reported always immunizing at preventive and follow-up visits, almost all (96.3%) at chronic illness visits, and a majority (78.6%) at acute care visits. As a result, none of the self reported immunization practices was significantly correlated with missed opportunities. CONCLUSIONS: Missed opportunities appear to be best predicted by motivational psychosocial factors and not by knowledge or perceived barriers. Self-reported immunization practices do not correspond to actual immunization behavior. PMID- 11897459 TI - Multistate analysis of factors associated with intimate partner violence. AB - BACKGROUND: Reports on prevalence estimates and risk factors of intimate partner violence (IPV) are limited in that they (1) focus on specific subgroup populations that are not representative of all women or (2) involve long questionnaires that are not useful as surveillance tools. OBJECTIVES: To report prevalence estimates and identify demographic and lifestyle factors associated with IPV in a large population-based sample of U.S. women using surveillance data. METHODS: Behavioral Risk Factor Surveillance System (BRFSS) data from eight U.S. states were analyzed individually and as a pooled sample (N=18,415). Multivariate logistic regression models were used to examine associations between IPV and the factors of interest. RESULTS: Factors consistently associated with IPV across the majority of states and in the pooled analysis included young age (pooled adjusted odds ratio [aOR], 3.07), single marital status (pooled aOR, 2.89), divorced/separated marital status (pooled aOR, 4.67), and annual household income <$25,000 (pooled aOR, 1.89). In addition, lack of health insurance, receipt of Medicaid, cigarette smoking, presence of children in the home, self reported fair/poor health, and frequent mental distress were associated with IPV after adjustment for covariates. CONCLUSIONS: This study provides population based estimates of IPV prevalence and factors associated with IPV using surveillance data. By pooling BRFSS data from individual states, the resulting large sample has adequate power to detect significant associations and has increased precision in the estimates of IPV risk. In addition, this study identifies high-risk populations to target for education and intervention programs and demonstrates the need for improved IPV surveillance. PMID- 11897461 TI - Pediatric deaths reported after vaccination: the utility of information obtained from parents. AB - BACKGROUND: The federally administered Vaccine Adverse Event Reporting System (VAERS) is a passive reporting system that receives domestic and foreign reports of adverse events that occur following immunization. This investigation explored whether routinely interviewing parents for follow-up of VAERS pediatric deaths would provide additional information important to vaccine safety. METHODS: The study was designed to follow up 100 consecutive pediatric deaths reported to VAERS by interviewing a parent and a healthcare provider (HCP) for each case. Several strategies contributed to successful follow-up. A standardized questionnaire was utilized to interview HCPs and parents. Overall and specific group frequencies (HCPs and parents) were calculated for each variable. McNemar's statistical tests of exact inference were calculated to assess whether there were statistically significant differences between HCP and parent knowledge by case for various variables. RESULTS: The median age of the cases was 4 months. Approximately half of the deaths were attributed to sudden infant death syndrome. In many instances, the information was equivalent in quality. For certain variables, such as knowledge of the child's position when found in distress, more parents than HCPs indicated that they knew the answer. CONCLUSIONS: Conducting parental and HCP follow-up for pediatric deaths reported to VAERS was resource intensive. In some instances, parents were more likely than HCPs to provide information regarding some important variables about the nature of the death. None of the additional information obtained from parents, however, provided a signal or confirmation of a causal link between the vaccine and death. PMID- 11897462 TI - Effectiveness of telephone support in increasing physical activity levels in primary care patients. AB - BACKGROUND: Physician counseling of patients to increase physical activity has had limited success in changing behavior. Providing organizational support to primary care providers and their patients may increase effectiveness. OBJECTIVE: This study evaluates the effectiveness of a telephone-based intervention to increase physical activity among patients who exercised <15 minutes daily and wanted to increase their physical activity over a 6-month period. DESIGN: This was a randomized controlled trial, conducted from 1997 to 1998, of 316 patients aged 18 to 65 who were recruited from a mailed health risk assessment. Baseline and 6-month post-intervention telephone assessments were conducted by telephone. SETTING: One family physician's patients in a suburban community. INTERVENTION: Three sessions of telephone-delivered motivational counseling. MAIN OUTCOME MEASURES: Physical activity score (11-item Physician-Based Assessment and Counseling for Exercise [PACE]) 6 months after the intervention. RESULTS: After adjusting for baseline exercise, there was a significantly higher level of self reported exercise among individuals randomized to the intervention at the 6-month follow-up. The mean level of activity at follow-up for the intervention group was a PACE score of 5.37, compared to 4.98 in the control group (p<0.05). In the secondary analysis, which was limited to individuals who received the intervention, the effect was stronger (PACE score of 5.58 compared to 4.94, p<0.013). CONCLUSIONS: Patients can be recruited using a health-screening questionnaire to receive a telephone-delivered behavioral intervention to successfully increase their physical activity levels. PMID- 11897463 TI - Assessing immunization registry data completeness in Bexar County, Texas. AB - BACKGROUND: Immunization information systems (or registries) are increasingly being used to promote and sustain high levels of vaccination coverage. However, the perception among many providers that registry data are too incomplete to be relied on when making immunization decisions has impeded the acceptance of registries. METHODS: To evaluate registry completeness, immunization coverage levels from the San Antonio Immunization Registry System (SAIRS) were compared with coverage levels derived from immunization records from 77 (37%) of the 210 clinics participating in the Vaccines for Children (VFC) program in 1998, 44 (21%) clinics in 1999, and 10 (5%) clinics in 2000. RESULTS: Clinic data indicated an average immunization coverage level for the 4:3:1 series of 39.8%. The overall coverage level for these clinics based on registry data was 64.1%. Registry-coverage levels for these clinics were < or =65% above the coverage levels based on clinic records. CONCLUSIONS: Immunization coverage levels based on SAIRS data were the same or higher than coverage levels based on clinic records. These data suggest that San Antonio's registry data were more complete than clinic records and may assist in changing provider perceptions regarding registry data completeness. PMID- 11897464 TI - Environmental factors associated with adults' participation in physical activity: a review. AB - BACKGROUND: Promoting physical activity is a public health priority, and changes in the environmental contexts of adults' activity choices are believed to be crucial. However, of the factors associated with physical activity, environmental influences are among the least understood. METHOD: Using journal scans and computerized literature database searches, we identified 19 quantitative studies that assessed the relationships with physical activity behavior of perceived and objectively determined physical environment attributes. Findings were categorized into those examining five categories: accessibility of facilities, opportunities for activity, weather, safety, and aesthetic attributes. RESULTS: Accessibility, opportunities, and aesthetic attributes had significant associations with physical activity. Weather and safety showed less-strong relationships. Where studies pooled different categories to create composite variables, the associations were less likely to be statistically significant. CONCLUSIONS: Physical environment factors have consistent associations with physical activity behavior. Further development of ecologic and environmental models, together with behavior-specific and context-specific measurement strategies, should help in further understanding of these associations. Prospective studies are required to identify possible causal relationships. PMID- 11897465 TI - Are biomarkers useful treatment aids for promoting health behavior change? An empirical review. AB - BACKGROUND: Nearly half of the leading causes of death in our society are attributable to behavioral risk factors. As such, it is critical that we continue to develop and refine effective interventions for health behavior change. Some researchers have suggested that using biomarkers to educate individuals about their health status and disease risk may be an effective strategy to promote behavior change. This tactic is also commonly employed by healthcare providers, but its empirical support is unclear. This article reviews the research literature to determine the effectiveness of using biomarker feedback to motivate and enable health behavior change. Potential limitations of this treatment strategy and issues requiring additional research are also discussed. METHODS: Articles were identified through PubMed (MEDLINE), PsychInfo, and the reference lists of pertinent manuscripts and book chapters. RESULTS: Eight published, randomized trials were identified that met criteria for review. The results of this work were mixed, but suggest that biological information conveying harm exposure, disease risk, or impaired physical functioning may increase motivation to change. Subsequent behavior change is also affected by the availability and intensity of concomitant treatment. CONCLUSIONS: Preliminary findings suggest that combining biomarkers with appropriate behavioral treatment may enhance health behavior change, but more research in this area is warranted. PMID- 11897466 TI - How the environment shapes physical activity: a transdisciplinary research agenda. PMID- 11897467 TI - Extending the methodology of the Committee on Clinical Preventive Service Priorities to HIV-prevention community planning. PMID- 11897468 TI - Physician roles in medicine-public health collaboration: future directions of the American Medical Association. PMID- 11897469 TI - In response to the October 2001 article entitled "Public health physicians, an endangered species." Public health physicians--not the only endangered species. PMID- 11897472 TI - Intracranial arachnoid cysts in children: a review of pathogenesis, clinical features, and management. AB - Arachnoid cysts are developmental anomalies that are most often diagnosed in childhood. They are often discovered as incidental findings found on imaging. Occasionally they may produce symptoms because of expansion or bleeding. There may be underlying maldevelopment of the cortex especially the temporal lobe. There is controversy regarding the role and the type of surgery indicated in its treatment. Recent descriptions of aphasia and attention-deficit disorders associated with these cysts indicate that we do not fully understand this entity. There is also no acceptable explanation for the male preponderance and increased incidence on the left side. The distribution, clinical features, treatment modalities, and some unusual syndromes associated with arachnoid cysts in children are discussed in this review. PMID- 11897473 TI - Meningitis and shunt infection caused by anaerobic bacteria in children. AB - This review describes the microbiology and management of meningitis and shunt infections caused by anaerobic bacteria in children. The predominant anaerobes recovered in meningitis are Bacteriodes spp., Bacteriodes fragilis, Fusobacterium spp., and Clostridium spp. Peptostreptococcus, Veillonella, Actinomyces, Propionibacterium acnes, and Eubacterium are less commonly isolated. The predisposing conditions for meningitis are acute or chronic middle-ear infection, sinusitis, pharyngitis, and pulmonary infections. In newborn and preterm infants the predisposing conditions are rupture of membranes, amnionitis, fetal distress, necrotizing enterocolitis, gastric perforation and subsequent ileus followed by bacteremia, aspiration pneumonitis and septicemia, infected ventriculoperitoneal or ventriculoatrial shunt, and complicating dermal sinus tract infections. Shunt infection with Propionibacterium spp. has been reported in children, especially in association with ventriculoauricular and ventriculoperitoneal shunts. Clostridium perfringens has been recovered from infants with a ventriculoperitoneal shunt. Multiple-organism meningitis was reported as a complication of ventriculoperitoneal and lumboperitoneal shunts that perforated the gastrointestinal tract. Early recognition and effective therapy are essential to recovery. Management of meningitis includes the use of antimicrobials effective against anaerobes that penetrate the blood-brain barrier. These include metronidazole, chloramphenicol, the combination of a penicillin and a beta lactamase inhibitor, and carbapenems. The treatment of shunt infection includes antimicrobial therapy and removal of the shunt. PMID- 11897474 TI - Language plasticity revealed by electroencephalogram mapping. AB - Reasoning is the result of the computations made by intelligent systems, for instance those in the brain. It is not an abstract concept because calculations performed by computations are very concrete transactions among the different central processing unit components. Entropy measurements are proposed here to disclose the plasticity of the cerebral processing associated with language comprehension in video game playing. It is also assumed that entropy may be evaluated from the correlation coefficients obtained for the game event-related activity calculated for the different electroencephalogram derivations in the 10/20 system. The brain mapping derived from these entropy measurements clearly demonstrates the reallocation of speech functions to right brain areas when the classic left language circuits are damaged during prenatal life. PMID- 11897476 TI - Epileptiform abnormalities in children with attention-deficit-hyperactivity disorder. AB - The proportion of children with attention-deficit-hyperactivity disorder and epileptiform abnormalities is compared with an historic control group of normal school-aged children. The medical records of 655 children 5-16 years of age referred to a single pediatric neurologist (M.S.) from January 1991 to December 1999 with school problems, behavior problems, or hyperactivity were retrospectively reviewed. Clinical criteria for attention-deficit-hyperactivity disorder were satisfied in 476 of these children. An electroencephalogram was obtained from 347 patients and coded as epileptiform in 6.1 +/- 1.3%, which is significantly higher (chi-square test, P < 0.025) than the prevalence rate of 3.5 +/- 0.6% observed in a study of 3,726 normal school-aged children. The epileptiform abnormality was present only with activation procedures in six of our patients (hyperventilation [n = 2] and photic stimulation [n = 4]). Only three of the 21 children with epileptiform abnormalities developed a seizure disorder in our cohort. We conclude that the prevalence rate of epileptiform abnormalities is greater in children with attention-deficit-hyperactivity disorder compared with that observed in normal school-aged children when hyperventilation and photic stimulation are used. However, the clinical utility of routine electroencephalography in the diagnosis of a comorbid seizure disorder in children with attention-deficit-hyperactivity disorder is limited. PMID- 11897475 TI - Oxidative stress in the human fetal brain: an immunohistochemical study. AB - Because accumulation of oxidative modification products seems to relate to aging and has not been fully studied in fetal brains, an immunohistochemical examination was performed on nine brains ranging from 22-40 weeks of gestation. These brains did not demonstrate lesions except hypoxic-ischemic changes. Advanced glycation end products and 4-hydroxynonenal are generally reported to be negative in neurons of normal young brains, but, in the present study, distinct positive immunoreaction was observed in neurons of fetal brains. Positive immunoreaction appeared earlier in the medulla oblongata than in the cerebrum, and 4-hydroxynonenal began to accumulate earlier than advanced glycation end products. As for glial cells, advanced glycation end products and 4 hydroxynonenal were positive in reactive astrocytes in mid- to late gestation. Because hypoxic-ischemic changes were observed in most of the patients, it is possible that oxidative stress caused by hypoxic-ischemic may be involved in the accumulation of these products in the fetal brain. 8-Hydroxy-2'-deoxyguanosine was negative even in patients demonstrating positive reaction for advanced glycation end products and 4-hydroxynonenal. In the fetal brain, DNA might be strongly protected from oxidative damage. 4-Hydroxynonenal is generally positive in the cytoplasm but was positive in the nucleus of immature neurons and glial cells in the present study, suggesting a unique metabolism of the fetal brain. PMID- 11897477 TI - Cutaneous blood flow and thermoregulation in Prader-Willi syndrome patients. AB - We examined adrenergic controlled cutaneous blood flow and temperature regulation in Prader-Willi syndrome. A body mass index was calculated for each participant. Thermal and laser Doppler finger probes were applied for continuous simultaneous surface temperature and capillary blood flow recording. Analysis with respect to group, age, body mass index, and genetic cause were performed. There were 32 patients (mean = 17.5 years of age) and five control subjects (mean = 15.6 years of age). There were no significant differences in mean ages or sex between groups. There was no significant difference in averaged blood flow measures with respect to group (P = 0.81), age (P = 0.16), body mass index (P = 0.54), or genetic identification (P = 0.81). There was no significant difference in average temperature measures as a function of group (P = 0.95), body mass index (P = 0.82), or genetic identification (P = 0.95). There was a significant difference in average temperature (P = 0.008) and trend in temperature change over time (P = 0.07) with respect to age for both patients and control subjects. Younger participants had higher average temperatures (30.6 degrees C vs 28.4 degrees C) in both study groups. We conclude that the central regulation and adrenergic control of cutaneous temperature and blood flow regulation in Prader-Willi syndrome at rest is not different from control subjects. These observations strengthen prior observations that a primary disturbance in parasympathetic autonomic regulation exists in Prader-Willi syndrome. PMID- 11897478 TI - Morphometric and neuropsychologic studies in children with arachnoid cysts. AB - Temporal lobe arachnoid cysts are common findings during brain imaging. Debate exists regarding whether they result from temporal lobe agenesis or are a malformation of the arachnoid matter. We measured temporal lobe volumes in five children with left middle cranial fossa arachnoid cysts using morphometric analysis of magnetic resonance imaging scans. Three patients had neuropsychologic testing, and two patients had positron emission tomography scanning. All patients had significantly smaller left temporal lobes compared with the right side. On neuropsychologic testing two patients had cognitive deficits suggestive of left temporal lobe dysfunction. Temporal lobes adjacent to arachnoid cysts are smaller and less metabolically active when compared with the temporal regions on the opposite side. Patients with middle cranial fossa arachnoid cysts should undergo careful assessment of temporal lobe structure and function before any therapeutic intervention. PMID- 11897479 TI - Episodic coma in a new leukodystrophy. AB - Among the leukodystrophies of a hypomyelinating nature, childhood ataxia with diffuse central nervous system hypomyelination exhibits the unique feature of rapid decrease in mental status after relatively minor head injuries or otherwise noncomplicated febrile illnesses. This article reports the case of a child with progressive spastic quadriparesis in whom unconsciousness developed repeatedly as a result of minor head trauma and required prolonged critical-care nursing. Although cognition is believed to be relatively preserved in this disorder, this child developed progressive cognitive decline. A detailed review of the literature is presented along with discussion of the potential mechanisms of neurologic deterioration. PMID- 11897480 TI - Pediatric phantom vision (Charles Bonnet) syndrome. AB - Visual symptomatology in childhood often presents diagnostic difficulties. Recurrent paroxysmal visual complaints, although typically associated with migraine, may also signal other disorders. We describe a 9-year-old partially sighted male with paroxysmal zoopsias resulting from Charles Bonnet syndrome. This condition is characterized by paroxysmal visual hallucinations occurring in patients with chronic visual impairment, akin to the phantom-limb phenomenon. This pediatric case is the fourth report of this condition. We have reviewed the other cases. PMID- 11897481 TI - Pyridoxal phosphate-responsive epilepsy with resistance to pyridoxine. AB - We present a female infant with seizures responsive to pyridoxal phosphate but that are resistant to pyridoxine. The mechanism by which pyridoxal phosphate controls seizures in this patient is unknown. Her seizures are perhaps not solely caused by pyridoxal phosphate deficiency. It is suggested that in addition to glutamic acid decarboxylase abnormality, the path from the absorption, transportation, phosphorylation, and oxidation of pyridoxine to pyridoxal phosphate in this patient might be defective. It should be considered whether pyridoxal phosphate can be the drug of choice instead of pyridoxine in treating patients suspected of pyridoxine-dependent epilepsy to reduce failure rate and further delay in seizure control. PMID- 11897482 TI - Osteoid osteoma presenting with focal neurologic signs. AB - Osteoid osteomas are benign bone tumors, most commonly located in the femur or tibia. In young children, these tumors can be extremely difficult to diagnose. They commonly present nonspecifically with gait disturbance and pain and characteristically respond well to mild pain relievers. We report two patients who presented with neurologic signs, including atrophy, weakness, and diminished deep tendon reflexes of the affected limb. These two patients demonstrate that osteoid osteomas of the lower extremities can present with neurologic signs, and proper diagnosis requires a detailed history and clinical awareness of this phenomenon. PMID- 11897483 TI - Pediatric granulomatous cerebral amebiasis: a delayed diagnosis. AB - We present four cases of cerebral amebae infection treated at our neurosurgical department. Patient 1 was a 12-year-old male with skin lesions of 2 years' progression involving the midface. He received a corticosteroid course, and, after that, he presented a right body hemiparesis. Patient 2 was a 5-year-old male, with a past surgical history of fibula fracture and osteomyelitis of 1-year evolution, associated with lesions of the surrounding skin that presented with partial seizures. Patient 3 was a 3-year-old female who presented with a stroke like episode and with partial seizures. Patient 4 was a 6-year-old male who had ulcerative lesions in the face of 1-year evolution. After a corticosteroid course, he presented with right-body hemiparesis. All patients were human immunodeficiency virus-negative and died 1 month or less after surgery because of progressive evolution of the disease. Histopathology revealed granulomatous amebic encephalitis. All patients revealed infection from Balamuthia mandrillaris (Leptomyxiidae). Treatment consisting of pentamidine, clarithromycin, fluconazole, and 5-fluorocytosine was ineffective. Although extremely uncommon, granulomatous amebic encephalitis should be considered in the differential diagnosis of cerebral lesions while nonspecific, associated granulomatous skin lesions support the diagnosis of amebiasis. PMID- 11897484 TI - Moyamoya, dystonia during hyperventilation, and antiphospholipid antibodies. AB - A 5-year-old Asian male presented with episodes of dystonia involving the right upper extremity during vigorous crying. He was diagnosed with moyamoya disease. Initial laboratory evaluation revealed positive anticardiolipin and antinuclear antibodies. PMID- 11897485 TI - Postmeningoencephalitic dysautonomia: report of one case. AB - We present a patient with postinfectious dysautonomia, which developed 2 weeks after a meningoencephalitic episode. The chief dysautonomic symptoms and signs included constipation, urinary dysfunction, anhydrosis, ptosis, and orthostatic hypotension. Neurophysiologic studies revealed no involvement of the somatic nervous system. The dysautonomia recovered gradually 3 months later with minimal supportive treatment. PMID- 11897488 TI - TNF receptor subtype signalling: differences and cellular consequences. AB - Tumour necrosis factor-alpha (TNF alpha) is a multifunctional cytokine belonging to a family of ligands with an associated family of receptor proteins. The pleiotropic actions of TNF range from proliferative responses such as cell growth and differentiation, to inflammatory effects and the mediation of immune responses, to destructive cellular outcomes such as apoptotic and necrotic cell death mechanisms. Activated TNF receptors mediate the association of distinct adaptor proteins that regulate a variety of signalling processes including kinase or phosphatase activation, lipase stimulation, and protease induction. Moreover, the cytokine regulates the activities of transcription factors, heterotrimeric or monomeric G-proteins and calcium ion homeostasis in order to orchestrate its cellular functions. This review addresses the structural basis of TNF signalling, the pathways employed with their cellular consequences, and focuses on the specific role played by each of the two TNF receptor isotypes, TNFR1 and TNFR2. PMID- 11897489 TI - Nitric oxide and the other cyclic nucleotide. AB - Nitric oxide (NO) participates in the regulation of the daily activities of cells as well as in cytotoxic events. Elucidating the mechanism(s) by which NO carries out its diverse functions has been the goal of numerous laboratories. In the cardiovascular system, evidence indicates that NO mediates its effects via an activation of soluble guanylyl cyclase (sGC). In other tissues, it is not clear if sGC is an exclusive target for NO or what the functions of cGMP might be. It is also unlikely that the diversity of NO actions is explained solely by changes in cGMP. This review focuses on the evidence that NO modulates cAMP signalling, with specific attention to the effects of NO on adenylyl cyclase (AC) as the target of NO regulation. PMID- 11897490 TI - A novel role for Gq alpha in alpha-thrombin-mediated mitogenic signalling pathways. AB - alpha-Thrombin activates several G-proteins including members of the Gq, Gi, and G12/13 families, although the physiological importance of these proteins is still not completely understood. We specifically investigated the role of Gq alpha in modulating alpha-thrombin-induced mitogenesis. In Gqa1 cells, a stable cell line expressing reduced amounts of Gq alpha, concentrations of alpha-thrombin (1 NIH unit/ml), which induce cell cycle reentry and progression into S phase in wild type IIC9 cells, do not stimulate phosphatidylinositol (PI) hydrolysis, the rapid early phase of ERK activity, and transit through G1 into S phase as quantified by cyclin-dependent kinase (CDK)4-cyclin D activity and [3H]thymidine incorporation. Interestingly, high concentrations of alpha-thrombin restore these activities and cell cycle progression into S phase. While, it is well documented that alpha thrombin-induced sustained ERK activity mediates important responses for transit through G1 into S phase, the importance of the rapid, Gq-dependent phase as a prerequisite for alpha-thrombin-mediated mitogenesis has not been appreciated. PMID- 11897492 TI - Luminal Ca(2+) and the activity of sarcoplasmic reticulum Ca(2+) pumps modulate histamine-induced all-or-none Ca(2+) release in smooth muscle cells. AB - We have studied histamine (HA)-evoked intracellular Ca(2+) release in single, freshly isolated myocytes from the guinea pig urinary bladder. Short applications of histamine (5 s) produced a thapsigargin (TG)-sensitive transient increase in intracellular calcium concentration ([Ca(2+)](i)). It was established that histamine and caffeine (Caff) released Ca(2+) from the same intracellular stores in these cells. Reducing the Ca(2+) content of internal stores by incubating cells with U-73343 or cyclopiazonic acid (CPA) inhibited the histamine-evoked Ca(2+) release in 69% and 60% of cells, respectively. Under these conditions, all cells released Ca(2+) in response to either caffeine or acetylcholine (ACh). However, decreasing internal Ca(2+) stores by removing external Ca(2+) inhibited histamine-induced Ca(2+) mobilization in only 22% of cells. A similar small fraction of cells was inhibited when sarcoplasmic reticulum (SR) Ca(2+) pumps were quickly blocked to avoid a significant reduction of luminal Ca(2+). In conclusion, lowering the luminal Ca(2+) content in combination with an impairment of the SR Ca(2+) pump activity significantly diminishes the ability of histamine to evoke an all-or-none intracellular Ca(2+) release. PMID- 11897491 TI - Modes of activation of mitogen-activated protein kinases and their roles in cepharanthine-induced apoptosis in human leukemia cells. AB - We previously showed that cepharanthine (CEP), a biscoclaurine alkaloid, induces caspase-dependent and Fas-independent apoptosis in Jurkat and K562 human leukemia cells. In the present study, we investigated the effect of CEP on three groups of human mitogen-activated protein kinases (MAPKs) in relation to CEP-induced apoptosis. CEP, at the concentration required for and at the time of induction of apoptosis, activated MAPKs p38 in both Jurkat and K562 cells and activated extracellular signal-regulated kinases (ERKs) only in K562 cells. However, CEP treatment did not trigger c-Jun NH(2)-terminal kinases (JNKs) activation. CEP increased the expression and phosphorylation levels of c-Jun and ATF-2 transcription factors. zVAD-fmk, a general caspase inhibitor, did not inhibit CEP triggered p38 activation in Jurkat and K562 cells or ERK activation in K562 cells. Unexpectedly, pretreatment with a specific p38 inhibitor, SB203580, promoted CEP-induced apoptosis and caspase activation in Jurkat and K562 cells, whereas pretreatment with an MEK-1 inhibitor PD98059 inhibited CEP-induced apoptosis and caspase activation in K562 cells. A selective tyrosine kinase inhibitor, herbimycin A, which completely inhibited CEP-triggered ERKs activation, clearly promoted CEP-induced c-Jun expression and phosphorylation. Our results suggest that each of the three groups of MAP family members is uniquely involved in the CEP-mediated signal cascades in two different leukemia cell lines for inducing/regulating caspase activation and DNA fragmentation. PMID- 11897493 TI - PKC epsilon is associated with myosin IIA and actin in fibroblasts. AB - Proteins coimmunoprecipitating with protein kinase C (PKC) epsilon in fibroblasts were identified through matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI TOF m/s). This method identified myosin IIA in PKC epsilon immunoprecipitates, as well as known PKC epsilon binding proteins, actin, beta'Cop and cytokeratin. Myosin is not a substrate for PKC epsilon. Immunofluorescence analysis showed that PKC epsilon is colocalised with actin and myosin in actomyosin stress fibers in fibroblasts. Inhibitors of PKC and myosin ATPase activity, as well as microfilament-disrupting drugs, all inhibited spreading of fibroblasts after passage, suggesting a role for a PKC epsilon-actin myosin complex in cell spreading. PMID- 11897494 TI - Protein inhibitor of activated signal transducer and activator of transcription (STAT)-1 (PIAS-1) regulates the IFN-gamma response in macrophage cell lines. AB - Macrophage cell lines exhibit different responses to IFN-gamma depending on their maturation stage. We investigated the mechanisms underlying the differential IFN gamma responsiveness in the less mature P388.D1 and in mature RAW264.7 cells. A reduction in the binding activity of the signal transducer and activator of transcription-1 (STAT-1) to different STAT binding elements (SBEs) was observed in P388.D1. This reduced binding activity was not due to an impaired STAT-1 activation. Studies on the expression of a negative regulator of cytokine signalling, protein-inhibiting activated STAT-1 (PIAS-1), showed that this protein was expressed constitutively at high levels in P388.D1. Forced expression of a PIAS-1 homologue, the Gu binding protein (GBP), inhibited the STAT-1 mediated gene activation in RAW264.7 cells, whereas a construct expressing the 5' portion of GBP in the antisense orientation reverts the IFN-gamma-resistant phenotype of P388.D1. Thus, our results indicate that PIAS-1 may account for the differential IFN-gamma responsiveness in macrophage cell lines at different stages of maturation. PMID- 11897495 TI - Effects of reactive oxygen species on actin filament polymerisation and amylase secretion in mouse pancreatic acinar cells. AB - The present study investigates the effect of reactive oxygen species (ROS) on actin filament reorganisation and its relevance to exocytosis in pancreatic acinar cells. Treatment of pancreatic acini with cholecystokinin (CCK-8) induced spatial and temporal changes in actin filament reorganisation with an initial depolymerisation of the apical actin barrier followed by an increase in the actin filament content in the subapical area leading to amylase release. Hydrogen peroxide (H(2)O(2)) increased actin filament content and potentiated the polymerizing effects of CCK-8 in these cells but abolished the disruption of the apical actin layer and amylase release induced by CCK-8. Similar to CCK-8, ROS generated by the oxidation of hypoxanthine (HX) with xanthine oxidase (XOD) induced an initial decrease in actin filaments located under the apical membrane followed by a smaller increase in the content of actin filaments in the subapical area. XOD-generated ROS are able to increase amylase release in pancreatic acini although combination with CCK-8 leads to abnormal exocytosis. We provide evidence that indicates that CCK-8- and ROS-induced actin reorganisation is entirely dependent on Ca(2+) mobilisation and independent of PKC activation. The regulation of the actin cytoskeleton by ROS might be involved in radical-induced cell injury in pancreatic acinar cells. PMID- 11897496 TI - Lithium blocks the PKB and GSK3 dephosphorylation induced by ceramide through protein phosphatase-2A. AB - The biochemical mechanism of apoptosis induced by ceramide remains still unclear, although it has been reported that dephosphorylation of PKB at Ser-473 may be a key event. In this article, we show that C(2)-ceramide (N-acetyl-sphingosine) induces the dephosphorylation of both protein kinase B (PKB) and glycogen synthase kinase-3 (GSK3) in cerebellar granule cells (CGC). We also show that lithium protects against the apoptosis induced by C(2)-ceramide by blocking the dephosphorylation of both kinases. Since lithium inhibits in vivo the observed protein phosphatase-2A (PP2A) activation induced by ceramide, we hypothesise that the neuroprotective action of lithium may be due to the inhibition of the PP2A activation by apoptotic stimuli. PMID- 11897497 TI - Developmental differences in B cell receptor-induced signal transduction. AB - We have compared early signaling events at various stages of B cell differentiation using established mouse cell lines. Clustering of pre-B cell antigen receptor (BCR) or BCR induced the tyrosine phosphorylation of various proteins in all cells, although the phosphorylation pattern differed. In spite of the pre-BCR-induced tyrosine phosphorylation, we could not detect an intracellular Ca(2+) signal in pre-B cells. However, co-clustering of the pre-BCR with CD19 did induce Ca(2+) mobilization. In contrast to the immature and mature B cells, where the B cell linker protein (BLNK) went through inducible tyrosine phosphorylation upon BCR clustering, we observed a constitutive tyrosine phosphorylation of BLNK in pre-B cell lines. Both BLNK and phospholipase C (PLC)gamma were raft associated in unstimulated pre-B cells, and this could not be enhanced by pre-BCR engagement, suggesting a ligand-independent PLC gamma mediated signaling. Further results indicate that the cell lines representing the immature stage are more sensitive to BCR-, CD19- and type II receptors binding the Fc part of IgG (Fc gamma RIIb)-mediated signals than mature B cells. PMID- 11897498 TI - Steroid hormone-growth factor interactions in proliferative controls of the mammary gland and breast cancer: a rapidly evolving perspective. PMID- 11897499 TI - Cellular expression of estrogen and progesterone receptors in mammary glands: regulation by hormones, development and aging. AB - At present, there is an extensive body of literature documenting the participation of estrogen receptors (ER) and progesterone receptors (PR) in mammary gene expression. Yet, the precise roles of these receptors in regulating mammary development, carcinogenesis and the growth of a subset of tumors still remain unclear. Mammary glands are composed of various cell types with different developmental potentials. Further, ultimately, that it is their mutual interactions which dictate the behavior of mammary epithelial cells. Therefore, to resolve the roles of ER and PR in normal mammary growth, differentiation and carcinogenesis, analyses for the expression of these receptors at the level of individual cell types is of paramount importance. Accordingly, in the present studies using immunolocalization techniques, we document the ontogeny and cellular distribution of ER and PR during mammary development and in response to ovarian hormones and aging. In addition, we discuss the potential biological significances of the expression patterns of ER and PR during various physiological states. We believe that the observations reported here should provide a conceptual framework(s) for elucidating the roles of ER and PR in normal and neoplastic mammary tissues. PMID- 11897500 TI - 17Beta-estradiol is carcinogenic in human breast epithelial cells. AB - The association found between breast cancer development and prolonged exposure to estrogen suggests that this hormone is of etiologic importance in the causation of this disease. In order to prove this postulate, we treated the immortalized human breast epithelial cells (HBEC) MCF-10F with 17beta-estradiol (E(2)) for testing whether they express colony formation in agar methocel, or colony efficiency (CE), and loss of ductulogenesis in collagen matrix, phenotypes also induced by the carcinogen benz[a]pyrene (BP). MCF-10F cells were treated with 0.0, 0.007, 70nM, or 0.25mM of E(2) twice a week for 2 weeks. CE increased from 0 in controls to 6.1, 9.2, and 8.7 with increasing E(2) doses. Ductulogenesis was 75 +/- 4.9 in control cells; it decreased to 63.7 +/- 28.8, 41.3 +/- 12.4, and 17.8 +/- 5.0 in E(2)-treated cells, which also formed solid masses or spherical formations lined by a multilayer epithelium, whose numbers increased from 0 in controls to 18.5 +/- 6.7, 107 +/- 11.8 and 130 +/- 10.0 for each E(2) dose. MCF 10F cells were also treated with 3.7 microM of progesterone (P) and the CE was 3.39 +/- 4.05. At difference of E(2), P does not impaired the ductulogenic capacity. Genomic analysis revealed that E(2)-treated cells exhibited loss of heterozigosity in chromosome 11, as detected using the markers D11S29 and D11S912 mapped to 11q23.3 and 11q24.2-25, respectively These results also indicate that E(2), like the chemical carcinogen BP, induces in HBEC phenotypes indicative of neoplastic transformation. PMID- 11897501 TI - Do estrogens always increase breast cancer risk? AB - The etiology of breast cancer is closely linked to the female hormone estrogen, with high life-time exposure being suggested to increase breast cancer risk [Nature 303 (1983) 767]. However, there appears to be a disparity between studies attempting to establish an association between high estrogen levels and breast cancer risk. This disparity becomes smaller by taking into consideration a timing factor, and we propose that estrogens can increase, decrease, or have no effect on breast cancer risk, depending on the timing of estrogen exposure. We further propose that the timing of estrogenic exposures may play at least as important a role in affecting breast cancer risk as life-time exposure. PMID- 11897502 TI - Roles of androgens in the development, growth, and carcinogenesis of the mammary gland. AB - Androgens influence the development and growth of the mammary gland in women. Treatment of animals and cultured cells with androgens has either inhibitory or stimulatory effects on the proliferation of mammary epithelia and cancer cells; the mechanisms for these dual functions are still not very clear and are discussed in this review. Epidemiological data suggest that, similar to increased estrogens, elevated androgens in serum may be associated with the development of breast cancer. Experiments in rodents have also shown that simultaneous treatment of androgen and estrogen synergizes for mammary gland carcinogenesis. Similar synergistic effects of both hormones have been observed for carcinogenesis of the uterine myometrium of female animals and for carcinogenesis of the prostate and deferens of males. There are also clinical and experimental indications for a possible association of elevated levels of both androgens and estrogens with the development of ovarian and endometrial cancers. A hypothesis is thus proposed that concomitant elevation in both androgens and estrogens may confer a greater risk for tumorigenesis of the mammary gland, and probably other female reproductive tissues than an elevation of each hormone alone. PMID- 11897503 TI - Steroid hormone mimics: molecular mechanisms of cell growth and apoptosis in normal and malignant mammary epithelial cells. AB - Anti-estrogen (anti-E2) therapy with E2 receptor antagonists has a significant benefit in women with breast cancer, but it may also increase the risk for developing hormone-independent breast cancer for which there is no therapy similar to that used in hormone-dependent breast cancer. Therefore, there is a significant interest in the development of compounds that may provide therapeutic benefit for hormone-independent breast cancer without untoward risks and adverse effects. The estrogen receptor (ER) modulators with both agonistic as well as antagonistic properties may, thus, be exploited for the development of the next generation of compounds for the prevention and/or treatment of breast cancer. In this article, we have discussed the clinical indications, risks, benefits and mechanisms of action of ER modulators and related compounds, particularly indole 3-carbinol (I3C), which may open new avenues for the prevention and/or treatment of breast cancer. PMID- 11897504 TI - Signaling pathways regulating aromatase and cyclooxygenases in normal and malignant breast cells. AB - Aromatase (estrogen synthase) is the cytochrome P450 enzyme complex that converts C(19) androgens to C(18) estrogens. Aromatase activity has been demonstrated in breast tissue in vitro, and expression of aromatase is highest in or near breast tumor sites. Thus, local regulation of aromatase by both endogenous factors as well as exogenous medicinal agents will influence the levels of estrogen available for breast cancer growth. The prostaglandin PGE(2) increases intracellular cAMP levels and stimulates estrogen biosynthesis, and our recent studies have shown a strong linear association between CYP19 expression and the sum of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) expression in breast cancer specimens. Knowledge of the signaling pathways that regulate the expression and enzyme activity of aromatase and cyclooxygenases (COXs) in stromal and epithelial breast cells will aid in understanding the interrelationships of these two enzyme systems and potentially identify novel targets for regulation. The effects of epidermal growth factor (EGF), transforming growth factor-beta (TGFbeta), and tetradecanoyl phorbol acetate (TPA) on aromatase and COXs were studied in primary cultures of normal human adipose stromal cells and in cell cultures of normal immortalized human breast epithelial cells MCF-10F, estrogen responsive human breast cancer cells MCF-7, and estrogen-unresponsive human breast cancer cells MDA-MB-231. Levels of the constitutive COX isozyme, COX-1, were not altered by the various treatments in the cell systems studied. In breast adenocarcinoma cells, EGF and TGFbeta did not alter COX-2 levels at 24h, while TPA induced COX-2 levels by 75% in MDA-MB-231 cells. EGF and TPA in MCF-7 cells significantly increased aromatase activity while TGFbeta did not. In contrast to MCF-7 cells, TGFbeta and TPA significantly increased activity in MDA-MB-231 cells, while only a modest increase with EGF was observed. Untreated normal adipose stromal cells exhibited high basal levels of COX-1 but low to undetectable levels of COX-2. A dramatic induction of COX-2 was observed in the adipose stromal cells by EGF, TGFbeta, and TPA. Aromatase enzyme activity in normal adipose stromal cells was significantly increased by EGF, TGFbeta and TPA after 24h of treatment. In summary, the results of this investigation on the effects of several paracrine and/or autocrine signaling pathways in the regulation of expression of aromatase, COX-1, and COX-2 in breast cells has identified more complex relationships. Overall, elevated levels of these factors in the breast cancer tissue microenvironment can result in increased aromatase activity (and subsequent increased estrogen biosynthesis) via autocrine mechanisms in breast epithelial cells and via paracrine mechanisms in breast stromal cells. Furthermore, increased secretion of prostaglandins such as PGE(2) from constitutive COX-1 and inducible COX-2 isozymes present in epithelial and stromal cell compartments will result in both autocrine and paracrine actions to increase aromatase expression in the tissues. PMID- 11897505 TI - Hormone/growth factor interactions mediating epithelial/stromal communication in mammary gland development and carcinogenesis. AB - Epithelial/mesenchymal interactions begin during embryonic development of the mammary gland and continue throughout mammary gland development into adult life. Stromal and epithelial growth factors that may mediate interactions between these compartments of the mammary gland are reviewed. Since mammogenic hormones are the primary regulators of mammary gland development, special consideration is given to hormonal regulation of growth factors in order to explore the integration of hormones and growth factors in the regulation of mammary gland growth and neoplasia. Examination of hormonal regulation of the fibroblast growth factor (FGF)-7/FGFR2-IIIb receptor system in the mammary gland reveals that mammogenic hormones differentially regulate the synthesis of stromal growth factors and their epithelial receptors. These effects serve to optimize the action of estrogen and progesterone on mammary gland development and illustrate that the ratio of these two hormones is critical in regulating this growth factor axis. The role of stromal/epithelial mitogenic microenvironments in modulating the genotype and phenotype of preneoplastic and neoplastic lesions by chemical carcinogens is discussed. Finally, changes in growth factor expression during mammary tumor progression are described to illustrate the relative roles that stromally-derived and epithelial-derived growth factors may play during progression to hormone independent tumor growth. PMID- 11897506 TI - Epidermal growth factor receptor (EGFR) transactivation by estrogen via the G protein-coupled receptor, GPR30: a novel signaling pathway with potential significance for breast cancer. AB - The biological and biochemical effects of estrogen have been ascribed to its known receptors, which function as ligand-inducible transcription factors. However, estrogen also triggers rapid activation of classical second messengers (cAMP, calcium, and inositol triphosphate) and stimulation of intracellular signaling cascades mitogen-activated protein kinase (MAP K), PI3K and eNOS. These latter events are commonly activated by membrane receptors that either possess intrinsic tyrosine kinase activity or couple to heterotrimeric G-proteins. We have shown that estrogen transactivates the epidermal growth factor receptor (EGFR) to MAP K signaling axis via the G-protein-coupled receptor (GPCR), GPR30, through the release of surface-bound proHB-EGF from estrogen receptor (ER) negative human breast cancer cells [Molecular Endocrinology 14 (2000) 1649]. This finding is consistent with a growing body of evidence suggesting that transactivation of EGFRs by GPCRs is a recurrent theme in cell signaling. GPCR mediated transactivation of EGFRs by estrogen provides a previously unappreciated mechanism of cross-talk between estrogen and serum growth factors, and explains prior data reporting the EGF-like effects of estrogen. This novel mechanism by which estrogen activates growth factor-dependent signaling and its implications for breast cancer biology are discussed further in this review. PMID- 11897507 TI - The role of mitogen-activated protein (MAP) kinase in breast cancer. AB - Mitogen-activated protein kinase (MAP kinase) cascades transmit and amplify signals involved in cell proliferation as well as cell death. These signal transduction pathways serve as an indicators of the intensity of trafficking induced by various growth factor, steroid hormone, and G protein receptor mediated ligands. Three major MAP kinase pathways exist in human tissues, but the one involving ERK-1 and -2 is most relevant to breast cancer. Peptide growth factors acting through tyrosine kinase containing receptors are the major regulators of ERK-1 and -2. Estradiol, progesterone, and testosterone can act non genomically via membrane associated receptors to activate MAP kinase as can various other ligands acting through heterotrimeric G protein receptors. Recent studies demonstrate that breast cancers frequently contain an increased proportion of cells with the activated form of MAP kinase. In estrogen receptor positive breast tumors, MAP kinase pathways can exert "cross talk" effects at the level of ER induced transcription as well as at the level of the cell cycle. Estradiol stimulates cell proliferation by mechanisms which involve activation of MAP kinase, either through rapid, non-transcription effects or by increasing growth factor production and consequently MAP kinase. Progesterone and androgens also stimulate MAP kinase through both of these two mechanisms. Strategies used to treat hormone dependent breast cancer appear to result in upregulation of MAP kinase activation. Direct experimental data demonstrate that the pressure of estradiol deprivation results in the upregulation of MAP kinase in breast cancer cells growing in tissue culture and as xenografts. A number of investigators have now studied the expression of activated MAP kinase in human breast cancer tissues by enzymatic assay and by immunohistochemical techniques. Approximately half of breast tumors express more activated MAP kinase than does the surrounding benign tissue. Studies show a trend toward higher MAP kinase activity in primary tumors of node positive than in node negative patients. However, larger numbers of patients must be studied for these results to achieve statistical significance. The up-regulation of MAP kinase activity does not represent mutations of Ras, but appears to result from enhancement of growth factor pathway activation. No data are yet available on the relationship between MAP kinase activation and apoptosis. Additional studies are now needed to determine the precise relationship between MAP kinase activation and tumor proliferation, apoptosis, and degree of invasiveness as well as on disease free and overall survival. PMID- 11897509 TI - State of the art in the delivery of photosensitizers for photodynamic therapy. AB - In photodynamic therapy, one of the problems limiting the use of many photosensitizers (PS) is the difficulty in preparing pharmaceutical formulations that enable their parenteral administration. Due to their low water solubility, the hydrophobic PS cannot be simply injected intravenously. Different strategies, including polymer-PS conjugation or encapsulation of the drug in colloidal carriers such as oil-dispersions, liposomes and polymeric particles, have been investigated. Although these colloidal carriers tend to accumulate selectively in tumour tissues, they are rapidly taken up by the mononuclear phagocytic system. In order to reduce this undesirable uptake by phagocytic cells, long-circulating carriers that consist of surface modified carriers have been developed. Moreover, considerable effort has been directed towards using other types of carriers to improve tumour targeting and to minimize the side effects. One of the approaches is to entrap PS into the lipophilic core of low-density lipoproteins (LDL) without altering their biological properties. The LDL receptor pathway is an important factor in the selective accumulation of PS in tumour tissue owing to the increased number of LDL receptors on the proliferating cell surface. Specific targeting can also be achieved by binding of monoclonal antibodies or specific tumour-seeking molecules to PS or by the coating of PS loaded carriers. PMID- 11897510 TI - Intraperitoneal photodynamic therapy in the Fischer 344 rat using 5 aminolevulinic acid and violet laser light: a toxicity study. AB - OBJECTIVE: Our study was designed to investigate 5-aminolevulinic acid (ALA) as a candidate for intraperitoneal photodynamic therapy (IP-PDT). The toxicity of IP PDT and the effects of IP-PDT on abdominal and pelvic organs, particularly the small intestine, were investigated after ALA administration and illumination with violet laser light. STUDY DESIGN AND RESULTS: The toxicity of IP-PDT was evaluated in Fischer 344 rats in two ways. In the first part of the study local PDT effects on the intestine were analyzed histologically. Violet laser light (lambda: 406-415 nm) was applied as a 2 cm diameter spot on the intestine 3 h after intraperitoneal (i.p.) administration of 50 mg/kg ALA. (A) Histological tissue samples were taken 0 min, 6 h and 1, 2 and 3 days after treatment (optical dose 3.2 J/cm(2)). Immediately after local PDT (3.2 J/cm(2), 50 mg/kg ALA) showed no effect on the intestine. However, 6 h post PDT there was complete destruction of the mesothelial lining and the outer (longitudinal) smooth muscle. Ganglion cells of the myenteric (Auerbach) plexus were also destroyed. The inner circular smooth muscle, the muscularis mucosa and the lamina propria were unharmed. Marked lymphectasia was present at this time. (B) To determine the threshold light dose of tissue destruction caused by PDT, different optical doses (1.6, 3.2, 6.4 J/cm(2)) were administered and histologic analysis of tissue samples were obtained 1 day post treatment. Destruction of the entire external musculature, submucosal structures and muscularis mucosa of the intestine at the illumination site could be observed above 1.6 J/cm(2) (50 mg/kg ALA). In the second part of the study whole peritoneal cavity PDT (WPC-PDT) was performed by illumination of the whole peritoneal cavity with 1.6 J/cm(2) violet light 3 h after ALA administration using different drug doses (200, 100 and 50 mg/kg). WPC-PDT showed lethal toxicity with a drug dose above 50 mg/kg ALA at 1.6 J/cm(2). The probable cause of death in the first 3 days after IP-PDT was rhabdomyolysis, whereas when death occurred at longer time intervals, megaintestine associated with significant damage could be observed; however, without perforation of the intestinal wall. CONCLUSION: In rats WPC-PDT with 50 mg/kg ALA, 1.6 J/cm2 at lambda=415 nm was the maximum tolerable light dose. This dose is likely to be above the threshold of destruction of ovarian cancer micrometastasis. PMID- 11897511 TI - UV-B-induced formation of reactive oxygen species and oxidative damage of the cyanobacterium Anabaena sp.: protective effects of ascorbic acid and N-acetyl-L cysteine. AB - Reactive oxygen species (ROS) are involved in the oxidative damage of the cyanobacterium Anabaena sp. caused by UV-B (280-315 nm) radiation. UV-B-induced overproduction of ROS as well as the oxidative stress was detected in vivo by using the ROS-sensitive probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH DA). Thiobarbituric acid reactive substances (TBARS) and fluorometric analysis of DNA unwinding (FADU) methods were adapted to measure lipid peroxidation and DNA strand breaks in Anabaena sp. Moderate UV-B radiation causes an increase of ROS production, enhanced lipid peroxidation and DNA strand breaks, yielding a significantly decreased survival. In contrast, the supplementation of UV-A in our work only showed a significant increase in total ROS levels and DNA strand breaks while no significant effect on lipid peroxidation, chlorophyll bleaching or survival was observed. The presence of ascorbic acid and N-acetyl-L-cysteine (NAC) reversed the oxidative stress and protected the organisms from chlorophyll bleaching and the damage of photosynthetic apparatus induced by UV-B significantly, resulting in a considerably higher survival rate. Ascorbic acid also exhibited a significant protective effect on lipid peroxidation and DNA strand breaks while NAC did not show a substantial effect. These results suggest that ascorbic acid exhibited significantly higher protective efficiency with respect to DNA strand breaks and survival than NAC while NAC appears to be especially effective in defending the photosynthetic apparatus from oxidative damage. PMID- 11897513 TI - Photomovement of the swarmers of the brown algae Scytosiphon lomentaria and Petalonia fascia: effect of photon irradiance, spectral composition and UV dose. AB - Photomovement measurements were carried out with swarmers of the brown algae Scytosiphon lomentaria (Lyngb.) Link and Petalonia fascia (O. F. Mull.) as a function of irradiance direction, photon irradiance, spectral composition and ultraviolet radiation (UVR, lambda=280-400 nm) dose. Swarmers from both species showed similar photomovement patterns: negative phototaxis occurred under photon irradiances of 10-90 micromol photons m(-2) s(-1), and no movement was observed at 190 micromol photons m(-2) s(-1). The translocational velocity measured between 10 and 90 micromol m(-2) s(-1) ranged from 100 to 200 microm s(-1). The accumulation of swarmers presented a peak at 450 nm (waveband of 50 nm), and smaller peaks at 400 and 500 nm; no effect was observed at wavelengths of 550 nm and above. The decline in phototactic index (an estimator of photomovement response) of swarmers was linearly correlated with the logarithm of UVR doses. These data were correlated with levels of natural solar radiation in the field. It is hypothesized that motility of swarmers could be a critical factor in the survival of these species under a scenario of increased UVR. PMID- 11897512 TI - Alteration of foliar flavonoid chemistry induced by enhanced UV-B radiation in field-grown Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii. AB - Chromatographic analyses of foliage from several tree species illustrate the species-specific effects of UV-B radiation on both quantity and composition of foliar flavonoids. Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii were field-grown under modulated ambient (1x) and enhanced (2x) biologically effective UV-B radiation. Foliage was harvested seasonally over a 3-year period, extracted, purified and the flavonoid fraction applied to a mu Bondapak/C(18) column HPLC system sampling at 254 nm. Total flavonoid concentrations in Quercus rubra foliage were more than twice (leaf area basis) that of the other species; Pseudotsuga menziesii foliage had intermediate levels and P. ponderosa had the lowest concentrations of total flavonoids. No statistically significant UV-B radiation-induced effects were found in total foliar flavonoid concentrations for any species; however, concentrations of specific compounds within each species exhibited significant treatment effects. Higher (but statistically insignificant) levels of flavonoids were induced by UV-B irradiation in 1- and 2-year-old P. ponderosa foliage. Total flavonoid concentrations in 2-year-old needles increased by 50% (1x ambient UV-B radiation) or 70% (2x ambient UV-B radiation) from that of 1-year-old tissue. Foliar flavonoids of Q. rubra under enhanced UV-B radiation tended to shift from early-eluting compounds to less polar flavonoids eluting later. There were no clear patterns of UV-B radiation effects on 1-year-old P. menziesii foliage. However, 2-year-old tissue had slightly higher foliar flavonoids under the 2x UV-B radiation treatment compared to ambient levels. Results suggest that enhanced UV-B radiation will alter foliar flavonoid composition and concentrations in forest tree species, which could impact tissue protection, and ultimately, competition, herbivory or litter decomposition. PMID- 11897514 TI - Application of very small force measurements in monitoring the response of sunflower to weak blue light. AB - A diaheliotropic response of sunflower, Helianthus annuus, following a short pulse of low intensity blue light to a small area of leaf surface was examined with the application of the very low-force-measurements technique (the order of magnitude of 10(-5) N). One leaf from a pair was illuminated with a low intensity blue-light-pulse and the force was recorded, generated by the stem of the plant tending to bend. A very low phototropic effect in response to blue light alone was observed which could be considerably enhanced by the application of background illumination with red light. Microelectrode measurements of the membrane potential of the mesophyll cells of the sunflower leaf showed hyperpolarization in response to a blue light pulse, observed very clearly under application of the red light background illumination. The hyperpolarization of the membrane potential was accompanied by acidification of extracellular compartments as monitored with a miniature pH-sensitive electrode, placed in the epidermis of the stem. The relatively short lag period between the hyperpolarization of the membrane potential and the decrease in pH suggests that the hyperpolarization is a direct effect of the blue light-induced proton extrusion. The acidification correlates with the light response, which suggests that acidification-induced stem wall loosening is responsible for the blue light induced bending. The examined mechanisms are discussed in terms of sun tracking by a sunflower. PMID- 11897515 TI - ATP synthesis in the disk membranes of rod outer segments of bovine retina. AB - ATP is synthesized on the disk membrane isolated from rod outer segments of the bovine retina. Together with a slow component which accounted for a constant rate of about 22 nmol ATP/min/mg of protein and which was due to the adenylate kinase activity, a fast component with a maximal activity of about 58 nmol ATP/min/mg of protein was measured at physiological calcium concentrations. This fast activity disappeared in the presence of the Ca(2+) ionophore A23187, was inhibited by vanadate or thapsigargin but not by oligomycin, suggesting that this ATP synthesis is due to the reversal functioning of the Ca(2+)-ATPase previously found on the disk membranes. PMID- 11897516 TI - Decreased survival of UV-irradiated Staphylococcus aureus in the presence of 8 methoxypsoralen in the post-irradiation plating medium. AB - For Staphylococcus aureus, the presence of 8-methoxypsoralen (8-MOP) in the post irradiation plating medium increased the lethal effect of far-UV light (FUV; approximately 254 nm) and of 8-MOP plus near-UV light (8-MOP+NUV; approximately 365 nm), an effect similar to that caused by acriflavine which inhibits DNA repair. In the repair-proficient strain, the presence of 8-MOP in the plating medium was almost as effective in inhibiting the repair of damage caused by FUV as that caused by 8-MOP photoadditions. Survival data obtained with Rec(-)-like and Uvr(-)-like strains suggest that 8-MOP in the plating medium, although possibly inhibiting recombination repair, was much more effective in inhibiting excision repair of FUV damage. Regarding 8-MOP+NUV treatment, 8-MOP in the plating medium had a lesser effect in the repair-deficient strains, differing from that observed after FUV treatment, which is consistent with the notion that different types of damage are caused by the two treatments. PMID- 11897517 TI - Bronchiolitis obliterans syndrome 2001: an update of the diagnostic criteria. PMID- 11897518 TI - Rejection is reduced in thoracic organ recipients when transplanted in the first year of life. AB - BACKGROUND: Infant heart transplant recipients have been reported to have decreased rates of rejection when clinical criteria are used for diagnosis. This study compares the rates of acute episodes of rejection in heart and lung transplant recipients transplanted in the first year of life to those of older recipients utilizing pathologic criteria. METHODS: Records of 100 consecutive lung transplant recipients (cystic fibrosis patients excluded) and 107 consecutive heart transplant recipients were reviewed with respect to: time to first rejection; total number; single versus multiple; and early (<90 days) versus late (>180 days) biopsy-proven rejection episodes. Rejection was defined as ISHLT biopsy Grade 3A or A2 for heart and lung transplant recipients, respectively. Biopsy and immunosuppression protocols were similar between groups. RESULTS: Kaplan-Meier analysis for freedom from rejection showed infant heart recipients were more often rejection-free (p = 0.004) as were infant lung recipients (p = 0.0001). Multivariate analysis revealed age at transplant as the most significant factor in predicting time to first rejection (age <1 year: risk ratio 0.19 [0.068-0.54] for lung transplant recipients and risk ratio 0.46 [0.27 0.78] for heart transplant recipients). Early rejection episodes occurred with less frequency in both the infant heart (19 of 63 [30%] versus 24 of 44 [50%]; p = 0.016) and lung (3 of 26 [12%] versus 63 of 74 [85%]; p = 0.001) groups. Late episodes of rejection were also less frequent in infant heart (4 of 53 [8%] versus 10 of 36 [28%], p = 0.016) and lung (0 of 23 [0%] versus 29 of 65 [45%]; p = 0.001) recipients. Multiple (> or =2) rejection episodes occurred less in infant heart (4 of 63 [6%] versus 9 of 41 [22%]; p = 0.037) and lung recipients (0 of 26 [0%] versus 17 of 74 [23%]; p = 0.003). CONCLUSIONS: These results demonstrate that age of <1 at time of thoracic transplantation confers significant protection from early, late and multiple episodes of acute rejection, as well as significantly greater freedom from rejection and time to first rejection. PMID- 11897519 TI - Success of lung transplantation without surveillance bronchoscopy. AB - BACKGROUND: No current evidence demonstrates improved survival or decreased rate of bronchiolitis obliterans syndrome (BOS) despite regularly scheduled fiberoptic bronchoscopy (FOB) with transbronchial biopsy and bronchoalveolar lavage (TBB/BAL) after lung transplantation. Reduced lung function detected with spirometry or oximetry in symptomatic and asymptomatic lung allograft recipients (LARs) may be a more appropriate indication for bronchoscopic sampling. HYPOTHESIS: Clinically indicated TBB/BAL without routine invasive surveillance sampling of the transplanted lung does not decrease survival or increase the rate of BOS in LARs. METHODS: We reviewed 91 consecutive LARs transplanted at Ochsner Clinic between January 1995 and December 1999. Clinical indications for FOB with TBB/BAL include 10% decline in forced expiratory volume in 1 second below baseline; 20% decrease in forced expiratory flow rate between 25% and 75% of the forced vital capacity; or unexplained respiratory symptoms, signs, or fever. Along with demographic and clinical data, 1-year and 3-year survival rates for these 91 LARs were compared with 5,430 LARs from the International Society for Heart and Lung Transplantation (ISHLT) Registry transplanted during the same 60 month period. Ten of the 91 patients did not survive to hospital discharge after transplantation. We divided the remaining 81 LARs into 2 subsets: Group A patients (n = 43) underwent zero to 1 TBB/BAL and Group B patients (n = 38) required more than 1 procedure. Demographic data, rejection, infection, and incidence of BOS were compared between groups. RESULTS: The 1-year and 3-year survival rates in the Ochsner LAR cohort were 85% and 73%, respectively, vs 72% and 57% in the ISHLT cohort p < 0.01. The relative risks of death in the Ochsner group at 1- and 3-years were 0.56 (0.35-0.91) and 0.66 (0.48-0.92), respectively, p < 0.05. The median (range) follow-up was 910 days (60-1,886) for Group A and 961 days (105-1,883) for Group B, p = not significant. We observed twice as many patients with cystic fibrosis and twice as many pneumonia episodes in Group B. The rate of acute rejection in each group was not statistically different. The cumulative incidence of BOS was increased in Group B at 1 year and at 3 years (5% and 56%) when compared with Group A (3% and 13%), p < 0.01. CONCLUSIONS: Based on the findings from this observational, single-institution study, clinically indicated TBB/BAL without routine surveillance sampling of the lung allograft is unlikely to pose greater risk than does regularly scheduled bronchoscopy after lung transplantation. PMID- 11897520 TI - Bronchiolitis obliterans syndrome in single lung transplant recipients--patients with emphysema versus patients with idiopathic pulmonary fibrosis. AB - Bronchiolitis obliterans syndrome (BOS) after lung transplantation is a disease of small airways that is currently graded according to a decline in forced expiratory volume in 1 second (FEV(1)) even in single lung transplant recipients in whom native diseased lung may influence lung physiology. The aim of this study was to evaluate the comparative changes in lung function and survival following the onset of BOS in patients with emphysema and patients with idiopathic pulmonary fibrosis (IPF) who have undergone single lung transplantation. We analyzed data from 31 single lung transplant recipients with emphysema and 25 with IPF who were at risk of BOS. There was no difference in the incidence of BOS between the 2 groups (10 patients with emphysema and 6 patients with IPF), but after the onset of BOS the patients with emphysema had a significantly greater median survival (18 months vs 8 months) despite a poorer mean FEV(1) (1.26 liter, 45% predicted vs 2.11 liter, 67% predicted) compared with the IPF group (p < 0.05) and this difference in lung function persisted at death (0.8 liter, 30% predicted vs 1.65 liter, 51% predicted) (p < 0.05). In summary the native lung physiology appears to influence lung function and therefore survival, and this may indicate that the classification of BOS should include disease-specific characteristics. PMID- 11897521 TI - Coronary risk stratification in patients with end-stage lung disease. AB - BACKGROUND: Significant coronary artery disease (CAD) has been a contraindication for listing patients for lung transplantation. We hypothesize that coronary risk stratification can help identify a sub-set of patients who need additional diagnostic tools and intervention. METHODS: We performed a retrospective review of 72 consecutive patients who underwent lung transplantation at our institution from 1995 to 2000. Further, a review of patients who are currently listed for transplantation yielded 48 patients. We then identified the various risk factors for CAD, the diagnostic tools used, and pre-operative intervention. Risk factors identified included smoking history, diabetes, hypertension, hypercholesterolemia, CAD, congestive heart failure, age >50, and arrhythmias. Based on these risk factors, the patients were then classified into 2 groups: low risk (< or =1 risk factors) and high risk (> or =2 risk factors). We identified the patients in each group who underwent coronary angiography (CA), those with angiographic evidence of CAD, and those who received pre-operative intervention. RESULTS: Of the 72 patients who underwent lung transplantation, 48 were identified as at high risk for CAD. Of these, 5 patients had CAD diagnosed before surgery using CA, and 1 patient received pre-operative intervention. Of the 48 patients currently on the lung transplant list, we identified 28 patients as high risk for CAD, 12 of whom were noted to have CA, and 2 of whom received pre operative intervention. CONCLUSIONS: Although CAD was once a contraindication for lung transplantation, pre-operative risk stratification allows identification of CAD with CA in a high-risk group. We believe that by using appropriate pre operative cardiac intervention, patients with severe CAD could successfully undergo lung transplantation. PMID- 11897522 TI - Donor interleukin-4 promoter gene polymorphism influences allograft rejection after heart transplantation. AB - BACKGROUND: The cytokine interleukin-4 (IL-4) is secreted mainly by activated T lymphocytes and characterizes the T-helper 2 (Th2) sub-type. In transplantation Th2 cells are believed to induce graft tolerance. Previous studies revealed that patients with a relatively high frequency of IL-4 producing helper T lymphocytes (HTL) before heart transplantation (HTX) had no or less rejection episodes compared with patients with a low frequency of IL-4 producing HTL. Three single nucleotide polymorphisms (SNPs) have been identified in the promoter region of the IL-4 gene, which influence promoter strength. We investigated whether there was a correlation between SNP genotypes in the IL-4 promoter and heart failure, and rejection after HTX. METHODS: Seventy HTX patients, 61 donors, and 36 controls were genotyped for the 3 SNPs by sequencing. RESULTS: Of the SNPs at 285 and -81, only the C and A alleles, respectively, were found in this study. Both alleles were found for the -590 SNP. No relation between patient genotype of the SNP at -590 and heart failure and rejection was found. However, incidence of rejection was significantly lower in patients that received a donor heart with the T-positive genotype compared with patients that received a heart from a T negative donor. Patients who had the T-negative genotype and received a heart from a T-positive donor, suffered significantly less from rejection than T negative patients that received a T-negative donor heart. This was not significant in the T-positive patient group. CONCLUSIONS: This indicates that IL 4 production within the donor heart and by cells from the donor is important for reducing incidence of episodes of rejection. PMID- 11897523 TI - Urapidil reduces elevated pulmonary vascular resistance in patients before heart transplantation. AB - BACKGROUND: Elevated pulmonary vascular resistance is a major limitation for heart transplantation. Urapidil is a centrally and peripherally acting anti hypertensive drug, able to decrease elevated pulmonary vascular resistance in patients with either chronic obstructive pulmonary disease or heart failure. Urapidil is available as an oral or intravenous drug. In this study, we evaluated the possible beneficial effects of intravenous urapidil in patients with reversible, elevated pulmonary vascular resistance who were scheduled for heart transplantation. METHODS: After approval by the Ethics Committee and written consent, 22 consecutive patients with end-stage heart failure and history of pulmonary vascular resistance >3 Wood units were enrolled into an open, prospective study. Using a (right ventricular ejection fraction) REF-Swan-Ganz catheter, hemodynamics were determined during administration of nitric oxide, and before and after 3 repeated intravenous applications of 10 mg urapidil. The treatment goal was reduction of pulmonary vascular resistance by at least 30%. RESULTS: Twenty-two patients were included to obtain complete data for 14 patients. Eight patients were not treated with urapidil: 7 patients had normal pulmonary vascular resistance at baseline, and 1 patient experienced moderate pulmonary edema before the study began. Two patients did not reach the treatment goal. In patients who responded to urapidil, the following hemodynamic changes were observed: decreased pulmonary vascular resistance (-48%), decreased transpulmonary gradient (20.0 to 13.7 mm Hg), decreased mean pulmonary arterial pressure (40 to 31 mm Hg), decreased systemic vascular resistance (-27%), mean arterial pressure (80 to 72 mm Hg), and increased right heart ejection fraction (21% to 27%). Heart rate remained unchanged. CONCLUSIONS: Intravenous urapidil lowered elevated pulmonary vascular resistance in patients before heart transplantation. In comparison with other vasodilative drugs, the major benefit of urapidil is its oral formulation. PMID- 11897524 TI - Nocardia infection in lung transplant recipients. AB - BACKGROUND: Nocardia is responsible for infection in both normal and immunocompromised hosts. Organ transplant recipients are increasingly recognized as a sub-group of immunocompromised patients in whom nocardia is an important pathogen. The frequency of nocardia in organ transplant recipients varies between 0.7% and 3%. Nocardia infection has largely been reported in heart, kidney and liver transplant recipients. Presentations of nocardia in lung transplant recipients have been restricted primarily to case reports. The present study reviews the clinical and epidemiologic characteristics of nocardia infection in lung transplant recipients at our institution. METHODS: A retrospective cohort study of 473 lung transplant recipients from January 1991 to November 2000 was done at a university hospital. Patient demographics, immunosuppressive regimen at the time of isolation of nocardia species, use of trimethoprim-sulfamethoxazole for Pneumocystis carinii prophylaxis, rejection episodes in the preceding 6 months, concurrent pathogens, site of infection, radiologic findings and treatment and outcome were recorded. RESULTS: Nocardia infection was found in 2.1% (10 of 473) of our lung transplant recipients. Median time of onset was 34.1 months after transplantation. Nocardia species included N farcinica in 30% (3 of 10), N nova in 30% (3 of 10), N asteroides complex in 30% (3 of 10) and N brasiliensis in 10% (1 of 10) of patients. Post-transplant diabetes was present in 50% (5 of 10) of patients. The primary indication for lung transplantation was emphysema in 40% (4 of 10). Native lung involvement was noted in 75% (3 of 4) of patients with single lung transplant. Breakthrough nocardia infection were noted in 6 patients who were receiving trimethoprim-sulfamethoxazole prophylaxis for P carinii pneumonia; all breakthrough isolates remained susceptible to trimethoprim sulfamethoxazole. Overall mortality was 40% (4 of 10). All patients (3 of 3) with infection due to N farcinica, except 1 (1 of 7) with infection due to other nocardia species, died. Seventy-five percent (3 of 4) of deaths were attributable to nocardia infection. CONCLUSIONS: Nocardia infection tended to involve the native lung in single lung transplant recipients. Trimethoprim-sulfamethoxazole for P carinii prophylaxis at the doses given was not protective against nocardiosis in these patients. Infection with N farcinica was associated with poor outcome. Thus, species identification and extended courses of antibiotics based on antimicrobial susceptibility testing are important in management of these patients. PMID- 11897525 TI - Zinc chloride-mediated reduction of apoptosis as an adjunct immunosuppressive modality in cardiac transplantation. AB - BACKGROUND: Zinc (Zn) blocks caspase-3 activation in cardiac allografts and therefore may synergistically decrease apoptosis along with cyclosporine (CsA), which inhibits mitochondrial release of cytochrome c. Simultaneous treatment of rat recipients of heterotopic heart transplants with zinc chloride (ZnCl(2)) thus may allow lower doses of CsA for immunosuppression. METHODS: PVG (RT1(c)) rat hearts were transplanted heterotopically into the abdomen of ACI (RT1(a)) rats. Group 1 (n = 15) rats received no treatment. Group 2 rats (n = 8) received 2 mg/kg/day CsA (sub-therapeutic dose) by oral gavage. Group 3 rats (n = 9) received 2 mg/kg/day oral CsA in addition to 1 mg/kg/day sub-cutaneous ZnCl(2) delivered by osmotic pump. All rats were imaged using Annexin V-bound (99m)Technetium ((99m)Tc-Annexin V) on post-operative Day 4 and subsequently killed. Annexin V avidly binds apoptotic cells in vivo. Region of interest per whole body (WB) data were calculated using the images. The allograft survival study was conducted with n = 11, 6, and 5 in control, CsA, and CsA+Zn groups, respectively. Finally, percentages of allografts that reached tolerance were measured in both CsA-only and CsA+Zn groups (n = 8 each). RESULTS: Zinc chloride had an additive effect with CsA on apoptotic blockade and graft survival. The regions of interest per WB uptake of (99m)Tc-Annexin V were 2.43% +/- 0.37%, 2.08% +/- 0.52%, and 1.49% +/- 0.29%*, and acute survivals were 6.4 +/- 1.7, 7.2 +/- 2.1, and 11.2 +/- 2.5* days for control, CsA, and CsA+Zn groups, respectively (*p < 0.001 vs controls). In addition, 87.5% of allografts became tolerant and survived for 90 days in the CsA+Zn group compared with only 37.5% in the CsA-only group (p = 0.049). CONCLUSION: Zinc-mediated reduction of apoptosis served as an effective adjunct immunosuppressive therapy to CsA in a rat model of cardiac transplantation. PMID- 11897526 TI - Molecular mechanisms of reduced sarcoplasmic reticulum Ca(2+) uptake in human failing left ventricular myocardium. AB - BACKGROUND: Human failing heart due to idiopathic dilated cardiomyopathy is associated with decreased sarcoplasmic reticulum Ca(2+) uptake. However, it is unknown as to which mechanism leads to this abnormality. METHODS: Immunodetectable sarcoplasmic reticulum proteins (phospholamban [PLB], phosphorylated PLB at serine-16 or threonine-17, calsequestrin and Ca(2+)-ATPase levels), the activities of Ca(2+)-calmodulin-dependent protein kinase and protein phosphatase and Ca(2+) uptake at varying Ca(2+) concentrations were determined in left ventricular specimens from the same 7 failing hearts (ejection fraction 20 +/- 2%) due to idiopathic dilated cardiomyopathy and 5 non-failing explanted control donor hearts. RESULTS: In failing hearts, compared with control donors, decreased maximal velocity and affinity of Ca(2+) uptake for Ca(2+) were found to be associated with reduced expression levels of Ca(2+)-adenosine triphosphatase (ATPase), PLB and phosphorylated PLB at serine-16, but not of calsequestrin and phosphorylated PLB at threonine-17. In contrast, protein phosphatase activity increased significantly and the activity and protein expression level of the delta isoform of Ca(2+)-calmodulin-dependent protein kinase remained unchanged in failing hearts compared with control donors. CONCLUSIONS: The impaired maximal velocity of sarcoplasmic reticulum Ca(2+) uptake may be due in part to reduced protein expression level of Ca(2+)-ATPase, whereas the reduced affinity may be due in part to the reduced ratio of Ca(2+)-ATPase to PLB and reduced PLB phosphorylation at serine-16 in failing hearts. The latter abnormality may be due in part to increased protein phosphatase activity. These results suggest that selective enhancement of Ca(2+) uptake into the sarcoplasmic reticulum by pharmaceutical agents, or by molecular tools that inhibit phosphatase activity, would be a valuable therapeutic approach for treating, or at least retarding, the process of heart failure. PMID- 11897527 TI - Inhibition of Na(+)/H(+) isoform-1 exchange protects hearts perfused after 6-hour cardioplegic cold storage. AB - OBJECTIVES: Cardiac ischemia-reperfusion activates Na(+)/H(+) exchange; excess Na(+) and the resulting Ca(2+) overload, through reverse Na(+)/Ca(2+) exchange, cause cellular injury and cardiac dysfunction. We postulated that inhibiting the Na(+)/H(+) isoform-1 exchanger would add to the protection of hearts after long term cold storage in acidic cardioplegic solution. METHODS: Guinea pig hearts were isolated and perfused at 37 degrees C with Krebs-Ringer's solution (KRS) and then switched to an acidic St. Thomas solution (STS) at 25 degrees C. Perfusion was stopped at 10 degrees C, and hearts were stored for 6 hours in STS at 3.4 degrees C. On reperfusion to 25 degrees C, hearts were perfused with KRS for 60 minutes. Hearts were divided into 4 groups: sham control (SHAM); eniporide (EPR, EMD96785) IV, 1 mg/kg given IV over 15 minutes before heart isolation; EPR intracoronary, 1 micromol/liter in STS given intracoronary after heart isolation; and EPR IV and intracoronary. RESULTS: Values at 60 minutes reperfusion (the percentage of control [100%] before cold storage) are given, respectively, for EPR IV, EPR intracoronary, and EPR IV and intracoronary vs drug-free SHAM (SEM, *p < 0.05 vs SHAM): 72% +/- 3%*, 65% +/- 3%*, and 81% +/- 2%* vs 55% +/- 3% for left ventricular pressure; 94% +/- 3%*, 96% +/- 5%*, and 102% +/- 2%* vs 81% +/- 3% for coronary flow; 60% +/- 2%, 58% +/- 3%, and 74%* +/- 3% vs 58% +/- 4% for cardiac efficiency; 106% +/- 2%*, 108% +/- 3%*, and 107% +/- 2%* vs 116% +/- 4% for percentage of O(2) extraction. Infarct size as percentage of ventricular weight was 20% +/- 3%*, 31% +/- 3%, and 6% +/- 2%* vs 35% +/- 3% (SHAM) after 60 minutes of reperfusion. CONCLUSIONS: Na(+)/H(+) isoform-1 exchanger inhibition, particularly if given IV before storage and intracoronary during cooling and rewarming, adds to the protection of cardioplegic solutions. PMID- 11897528 TI - The optimal pressure for initial flush with UW solution in heart procurement. AB - OBJECTIVE: University of Wisconsin (UW) solution is widely used in organ preservation. Some investigators have reported that high pressure during initial flush with UW solution may induce vasoconstriction and endothelial damage, because of its high potassium content and high viscosity. However, using lower pressure during the initial flush may lead to irregular distribution of the solution and incomplete flushing of blood components from coronary vascular beds. This experimental study evaluated the effects of a range of initial flush pressures during heart procurement, followed by orthotopic transplantation of the graft after 12 hours of preservation. MATERIALS AND METHODS: Twelve pairs of adult mongrel dogs, weighing 9 to 14 kg, formed the recipient-donor combinations. After determining hemodynamic status by measuring cardiac output, left ventricular pressure (LVP), and maximum positive and negative change in LVP (+/ LVdP/dt), donor hearts were excised. Coronary vascular beds were flushed with 4 degrees C UW solution at a pressure of 60 mm Hg in the low-pressure group (n = 6) and at 120 mm Hg in the high-pressure group (n = 6). After 12 hours of cold preservation, orthotopic transplantation was performed using cardiopulmonary bypass (CPB). The hemodynamics of the transplanted graft were assessed by comparing recovery rates (%) from donor hearts 2 hours after weaning from CPB. Endothelin-1 (ET-1) levels were measured in the blood obtained from the coronary sinus 30 minutes after reperfusion. The transplanted grafts were then harvested for histologic study and measurement of adenosine triphosphate (ATP) content. RESULTS: Cardiac output, LVP, LVdP/dt and myocardial tissue ATP content were significantly better (p < 0.05) in the high-pressure group than in the low pressure group. We found no significant differences in ET-1 levels between the groups. Transmission electron microscopic findings revealed that degeneration of the mitochondria was less extensive in the high-pressure group than in the low pressure group. We observed no obvious ultrastructural damage to the endothelial cells in either group. CONCLUSION: When using UW solution in heart procurement, high pressure is better to completely wash out the blood components and distribute the solution. PMID- 11897529 TI - Mycobacterium abscessus empyema in a lung transplant recipient. AB - Non-tuberculous mycobacteria (NTM) have emerged as important pathogens in organ transplant recipients. Because NTM pulmonary infections vary in their clinical and radiographic presentations, heightened clinical suspicion is necessary for accurate diagnosis. We report a case of Mycobacterium abscessus empyema in a lung transplant recipient. Repeated attempts at identifying the organism from a variety of clinical specimens led to the correct diagnosis and treatment. PMID- 11897530 TI - Long-term survival despite early loss of graft function after single lung transplantation for pulmonary fibrosis. AB - We report a patient who received a single, left lung transplantation for idiopathic pulmonary fibrosis. The effect of the graft on pulmonary improvement was only temporary, because the patient developed obliterative bronchiolitis (OB), resulting in complete destruction of the graft. The patient, however, remains alive 6 years after OB was diagnosed, apparently as a consequence of native lung improvement with triple-immunosuppressive medicine. This case is of interest for several reasons: first, it shows that pulmonary fibrosis may respond to intensive immunosuppressive therapy; second, it demonstrates that ventilation scintigraphy is useful in addition to pulmonary function tests in estimating the actual function of the graft after single lung transplantation; and third, it appears that the gradation of bronchiolitis obliterans syndrome (BOS) after single lung transplantation may overestimate the true function of the transplant. PMID- 11897531 TI - Alternative approach for use of a left ventricular assist device with a thrombosed prosthetic valve. AB - Implantation of a left ventricular assist device is problematic in patients with prosthetic heart valves, due to an increased risk of thrombosis with embolization. This report describes the use of a bovine pericardial patch to close the aortic outflow tract in a patient with a mechanical aortic valve and end-stage cardiomyopathy who required urgent left ventricular assist support. A successful outcome suggests that this technique may be of value in treating similar patients. PMID- 11897532 TI - Electrophysiology of radiculopathies. AB - The anatomy, pathophysiology, and clinical evaluation of radiculopathies are discussed. Defining whether root injury is present and which roots are involved can be difficult but critical for patient management. In conjunction with clinical and radiological information, studies that establish physiological abnormalities of roots should be helpful and important. Clinical neurophysiological studies for radiculopathies are performed frequently but have yet to achieve a universally accepted role in the evaluation of these patients. Electrophysiological techniques for the evaluation of radiculopathies are reviewed. Needle electromyography is the best established of these procedures but has the disadvantage of requiring injury to motor fibers of both a certain degree and distribution. Nerve conduction studies may rarely be abnormal in radiculopathies but are needed to be certain other conditions that may produce similar symptoms and signs are not present. H reflexes and F waves probably have roles in the evaluation of radiculopathies but published reports about F waves in radiculopathies have been marred by inadequate methodology. There is evidence based on large series of patients that somatosensory evoked potentials can be helpful for evaluating patients with multilevel injury such as spinal stenosis, patients where electrophysiological studies may have their greatest clinical utility. Further work using either electrical stimulation with needles or magnetic stimulation of roots seems warranted. The demonstration of meaningful electrophysiological changes with activities that reproduce radicular symptoms may be a promising experimental approach. Available information does not necessarily answer critical questions about the role of electrophysiology in patients with radiculopathies. This cannot be done using analyses based on current ideas about evidence based medicine given the absence of a 'gold standard' for defining radiculopathies as well the absence of blinded studies. The available information provides strong arguments for further investigations evaluating different clinical neurophysiological techniques in the same patient, and for evaluating the value of these techniques by concentrating on their clinical import. PMID- 11897533 TI - Electrophysiological assessment of the effect of intrathecal baclofen in spastic children. AB - OBJECTIVES: To evaluate the effect of intrathecal baclofen in a group of spastic children using electrophysiological procedures described in adults. METHODS: Six children (aged 1-14 years) with severe spasticity of various aetiologies underwent transcranial magnetic stimulation, H reflex and flexor reflex studies before and after intrathecal injection of baclofen. Ashworth scale was used for clinical evaluation of spasticity. RESULTS: Motor evoked potentials, present in two patients before baclofen, were preserved after injection. Before baclofen, H reflex was present in 5 patients (H(max)/M(max) from 0.23 to 0.84) and absent in one who had infantile neuroaxonal dystrophy. After baclofen, it was absent in 4 patients and markedly reduced in one. Surface of flexor reflex significantly decreased after baclofen (P=0.01), while threshold significantly increased (P=0.003). CONCLUSIONS: In spastic children, the action of baclofen on spinal pathways may be quantified by the same electrophysiological procedures as in adults. This approach may contribute to select optimal dosage. PMID- 11897534 TI - Pseudo-bilateral hand motor responses evoked by transcranial magnetic stimulation in patients with deep brain stimulators. AB - OBJECTIVES: In 3 of 5 patients with dystonia and bilaterally implanted deep brain stimulating electrodes, focal transcranial magnetic stimulation (TMS) of one motor cortex elicited bilateral hand motor responses. The aim of this study was to clarify the origin of these ipsilateral responses. METHODS: TMS and electrical stimulation of corticospinal fibres by the implanted electrodes were performed and the evoked hand motor potentials were analysed. RESULTS: In comparison with responses elicited by contralateral motor cortex stimulation, ipsilateral responses were smaller in amplitude (3.0+/-1.4 versus 5.8+/-1.5 mV), had shorter peak latencies (first negative peak: 20.9+/-0.8 versus 25.1+/-0.4 ms) and were followed by a shorter-lasting silent period (46+/-4 versus 195+/-35 ms). Ipsilateral responses following TMS had similar peak latencies to responses elicited subcortically by deep brain stimulation (DBS) (20.4+/-0.9 ms). CONCLUSIONS: Hand motor responses ipsilateral to TMS result from a subcortical activation of corticospinal fibres, via the implanted electrode in the other hemisphere, secondary to currents induced by TMS in subcutaneous wire loops that underlie the magnetic coil. Studies of TMS in patients with DBS have to take this potential source of confounding into account. PMID- 11897536 TI - Non-invasive monitoring of functionally distinct muscle activations during swallowing. AB - OBJECTIVES: Dysphagia is an important consequence of many diseases. As some of the muscles of deglutition tend to be deep to the surface, needle electrodes are typically used, but this limits the number of muscles that can be simultaneously recorded. Since control of swallowing involves central pattern generators (CPGs) which distribute commands to several muscles, monitoring several muscles simultaneously is desirable. Here we describe a novel method, based on computing the independent components (ICs) of the simultaneous sEMG recordings (Muscle Nerve Suppl 9 (2000) 9) to detect the underlying functional muscle activations during swallowing using only surface EMG (sEMG) electrodes. METHODS: Seven normal subjects repeatedly swallowed liquids of varying consistency while sEMG was recorded from 15 electrodes from the face and throat. Active areas of EMG were excised from the recordings and the ICs of the sEMG were calculated. RESULTS: The ICs demonstrated less swallow-to-swallow variability than the raw sEMG. The ICs, each consisting of a unique temporal waveform and a spatial distribution, provided a means to segregate the complex sequence of muscle activation into rigorously defined separate functional units. The temporal profiles of the ICs and their spatial distribution were consistent with prior needle EMG studies of the cricopharyngeal, superior pharyngeal constrictor, submental and possibly arytenoid muscles. Other components appeared to correspond to EKG artifact contaminating the EMG recordings, laryngeal excursion, tongue movement and activation of the buccal and/or masseter musculature At least two of the components were affected by the consistency of the liquids swallowed. Re-testing one subject a week later demonstrated good intertrial reliability. CONCLUSIONS: We propose that the ICs of the sEMG provide a non-invasive means to assess the complex muscle sequence activation of deglutition. PMID- 11897535 TI - Can electrophysiology differentiate polyneuropathy with anti-MAG/SGPG antibodies from chronic inflammatory demyelinating polyneuropathy? AB - OBJECTIVES: Patients with polyneuropathy and antibodies to myelin-associated glycoprotein (MAG) and sulphated glucuronyl paragloboside (SGPG) differ from chronic inflammatory demyelinating polyneuropathy (CIDP) because of a slower, progressive course, symmetrical and predominantly sensory involvement of legs, predominantly distal slowing of motor conductions, and poorer response to therapy. We studied whether a wide set of electrophysiologic parameters may differentiate these two neuropathies. METHODS: We reviewed the electrophysiological studies of 10 patients with anti-MAG/SGPG antibodies and 22 with CIDP examining: (1) motor conduction velocity and distal compound muscle action potential amplitude; (2) conduction block (CB) and temporal dispersion; (3) distal motor latency and terminal latency index (TLI); (4) F wave and proximal conduction time; and (5) sensory conduction and occurrence of abnormal median with normal sural sensory potential. RESULTS: Anti-MAG/SGPG neuropathies showed: (1) more severe involvement of peroneal nerves; (2) more frequent disproportionate distal slowing of motor conductions (TLI< or =0.25) and absent sural potential, and (3) no CB. However 3/22 CIDP patients also had at least two nerves with TLI< or =0.25 and no CB. CONCLUSIONS: Electrophysiologic findings suggest in anti-MAG/SGPG neuropathy a length-dependent process with a likely centripetal evolution. A disproportionate slowing of conduction in distal segments of motor nerves suggests the diagnosis of anti-MAG/SGPG neuropathy, although it is not pathognomonic. PMID- 11897537 TI - Time-related changes of excitability of the human motor system contingent upon immobilisation of the ring and little fingers. AB - OBJECTIVES: To examine possible changes of excitability of the human motor system contingent upon immobilisation of two hand fingers. METHODS: Two series of 5 transcranial magnetic stimulation (TMS) sessions were carried out on different days (1, 2, 3, 4, and 7). In one series (fingers immobilised, FI), subjects wore for 4 days a device that kept immobilised the left fourth and fifth finger. In the other series (fingers free, FF), no constraining device was used. Focal TMS was applied over the right motor cortex and motor evoked potentials (MEPs) were recorded from left abductor digiti minimi (immobilised) and first dorsal interosseus (non-immobilised) muscles. Intensities of 10, 30, and 50% above the resting motor threshold (rMT), were used. RESULTS: In FI series, rMT for both muscles showed significant increase on days 3, 4, and 7 with respect to day 1. At high stimulation intensity a clear decrease of MEPs amplitude was observed on days 3 and 4 for both muscles. Since no time-related changes of peripheral (M wave) and spinal (F-wave) excitability were noted, MEPs and rMT changes are likely to have a cortical origin. In FF series, no changes of excitability were detected. CONCLUSIONS: Sensorimotor restriction of two fingers induces an early decrease of excitability, possibly at cortical level, which involves not only the immobilised muscle but also muscles with purportedly overlapping neural representations. PMID- 11897538 TI - Inter- and intra-individual variability of paired-pulse curves with transcranial magnetic stimulation (TMS). AB - OBJECTIVES: Previous studies have evaluated the variability of motor thresholds (MTs) and amplitude of motor-evoked potentials (MEPs) to transcranial magnetic stimulation (TMS) within and across individuals. Here we evaluate the reproducibility and inter-hemispheric variability of measures of cortical excitability using the 'conventional' paired-pulse (PP) TMS technique. METHODS: We studied PP curves of the left and right hemisphere in 10 healthy subjects on two separate days 2 weeks apart. The inter-stimulus intervals studied were 1, 3, 6, 8, 10 and 12 ms with the conditioning stimulus being 80% of the resting MT, and a single test stimulus producing MEPs of approximately 0.8 mV peak-to-peak amplitude. RESULTS: As a group, the PP curves of the left and right hemispheres, and of Day 1 and Day 2 were not significantly different. The intracortical inhibition (ICI), but not the intracortical facilitation, showed a good correlation across days within the individuals. CONCLUSIONS: Cortical excitability, particularly ICI, measured by PP TMS shows no inter-hemispheric asymmetry and is reproducible within individuals. PMID- 11897539 TI - Semantic bias, homograph comprehension, and event-related potentials in schizophrenia. AB - OBJECTIVES: It is controversial whether a semantic processing bias for strong associates is present in schizophrenia, and unknown whether the language abnormalities observed in schizophrenia can be attributed to dysfunctions early or late in cognitive processing. Combined behavioral and event-related potential (ERP) data can indicate the nature and timing of such abnormalities. METHODS: Sensibility judgements of dominant and subordinate homograph sentences were measured in 12 schizophrenia patients and 13 normal controls. ERPs were recorded to the disambiguating sentence-ending word. RESULTS: All subjects showed greatest misinterpretation of subordinate homograph sentences, but schizophrenia patients particularly misinterpreted these sentence types. For control subjects, subordinate homograph sentences that were classified as nonsensical showed greater N400 than those classified as sensible. By contrast, the N400 of patients was large, regardless of the sensibility judgement--patients' brains initially responded to all subordinate sentences as if nonsensical. These data are consonant with a semantic bias. However, the patients' N400 to dominant homograph sentence endings was also larger than that of controls, a finding not consonant with a semantic bias. CONCLUSIONS: The behavioral results indicate a selective comprehension abnormality in schizophrenia dependent on the content of verbal memory. The ERP results suggest a pervasive contextual memory failure. A semantic activation decay model is proposed to explain these results. PMID- 11897540 TI - Temporal resolution in young and elderly subjects as measured by mismatch negativity and a psychoacoustic gap detection task. AB - OBJECTIVES: The speech understanding difficulties of elderly persons with age related hearing loss may be related in part to reduced auditory temporal resolution. To investigate the effects of aging on temporal resolution, the electrophysiological and psychoacoustic detection thresholds for a very short silent gap within a pure tone were determined, and the relation between the two test results was examined. METHODS: Behavioral gap detection thresholds were determined in 10 young and 10 elderly normally hearing subjects using an adaptive test procedure. To elicit mismatch negativity (MMN), deviant stimuli with gap durations varying from 6 to 24 ms in 3 ms steps were presented in separate test blocks. RESULTS: There were no significant differences in the psychoacoustic gap detection thresholds between young and elderly subjects. In contrast, longer gaps were needed to elicit MMN in elderly subjects. They also had significantly reduced MMN peak amplitudes, increased MMN peak latencies, a significantly smaller P2 amplitude and longer P2 latency in their responses to the standard stimulus when compared to the same measures in young subjects. CONCLUSIONS: Processing of basic temporal stimulus features in elderly subjects is considerably more reduced at the preattentive level (as indicated by MMN), than when attention is directed to the task (as indicated by the psychoacoustic results). PMID- 11897541 TI - Maturation of human central auditory system activity: separating auditory evoked potentials by dipole source modeling. AB - OBJECTIVES: Previous studies have shown that observed patterns of auditory evoked potential (AEP) maturation depend on the scalp location of the recording electrodes. Dipole source modeling incorporates the AEP information recorded at all electrode locations. This should provide a more robust description of auditory system maturation based on age-related changes in AEPs. Thus, the purpose of this study was to evaluate central auditory system maturation based dipole modeling of multi-electrode long-latency AEPs recordings. METHODS: AEPs were recorded at 30 scalp-electrode locations from 118 subjects between 5 and 20 years of age. Regional dipole source analysis, using symmetrically located sources, was used to generate a spatio-temporal source model of age-related changes in AEP latency and magnitude. RESULTS: The regional dipole source model separated the AEPs into distinct groups depending on the orientation of the component dipoles. The sagittally oriented dipole sources contained two AEP peaks, comparable in latency to Pa and Pb of the middle latency response (MLR). Although some magnitude changes were noted, latencies of Pa and Pb showed no evidence of age-related change. The tangentially oriented sources contained activity comparable to P1, N1b, and P2. There were various age-related changes in the latency and magnitude of the AEPs represented in the tangential sources. The radially oriented sources contained activity comparable to the T-complex, including Ta, and Tb, that showed only small latency changes with age. In addition, a long-latency component labeled TP200 was observed. CONCLUSIONS: It is possible to distinguish 3 maturation groups: one group reaching maturity at age 6 and comprising the MLR components Pa and Pb, P2, and the T-complex. A second group that was relatively fast to mature (50%/year) was represented by N2. A third group was characterized by a slower pattern of maturation with a rate of 11 17%/year and included the AEP peaks P1, N1b, and TP200. The observed latency differences combined with the differences in maturation rate indicate that P2 is not identical to TP200. The results also demonstrated the independence of the T complex components, represented in the radial dipoles, from the P1, N1b, and P2 components, contained in the tangentially oriented dipole sources. PMID- 11897542 TI - Habituation of the auditory evoked field component N100m and its dependence on stimulus duration. AB - OBJECTIVES: In the current study we investigated the habituation of the neuromagnetic auditory evoked field (AEF) component N100m and its dependence on stimulus duration. METHODS: Fifteen subjects were stimulated monaurally in 3 blocks of 210 trials with 1000 Hz tones of different duration (50, 100, 200 ms) and in a single block of 70 trials with 1200 Hz tones. The order of blocks was counterbalanced within the total experiment. Tones were separated by a stimulus onset asynchrony of 2000 ms. AEF were recorded over both hemispheres by means of a 31 channel system (Philips) on two different days. The AEF were compared within and between the blocks and between the hemispheres. Additionally, the effects of block order were analyzed. RESULTS: In the course of the experiment a pronounced decrease of N100m mean global field power (MGFP) and an increase of its latency were observed. While the order of blocks clearly affected the degree of habituation, stimulus duration did not have any influence on it. Within the blocks, habituation also had an impact on dipole location in inferior-superior direction. The application of the 1200 Hz stimulus led to a slight response recovery. The change in tone pitch affected the dipole orientation, as an indicator for the tonotopic organization of the auditory cortex. PMID- 11897543 TI - Ritanserin, a serotonin-2 receptor antagonist, improves ultradian sleep rhythmicity in young poor sleepers. AB - OBJECTIVES: To determine the effect on sleep electroencephalographic (EEG) activity of ritanserin, a serotonin-2 (5-HT2) receptor antagonist in young poor sleepers. METHODS: Eight male subjects underwent two randomized night studies after receiving either a placebo or 5 mg ritanserin administered in the morning. The overnight variations in the delta (0.5-4.0 Hz) and sigma (12.25-15.0 Hz) frequency bands were characterized using a peak analysis which provided a quantitative evaluation of the time-courses in EEG activity. RESULTS: In subjects under ritanserin, slow wave sleep duration and the number of non-rapid eye movement (NREM)-REM sleep cycles were significantly enhanced (P<0.01). The number of peaks in delta activity occurring in the normal 80-120 min range was significantly (P<0.05) increased. Using a delta peak analysis, 4 periods containing or not a significant peak were identified in each subject. A significant increase in delta activity was observed in the areas under the averaged curves during the second and the third periods (P<0.05), while sigma activity decreased under ritanserin during the first, second and third periods (P<0.05). CONCLUSIONS: These results demonstrate that ritanserin increases delta activity, possibly by opposing the inhibitory control of 5-HT2 receptor family. It restores sleep ultradian rhythmicity and improves sleep quality in young poor sleepers. PMID- 11897544 TI - Electroencephalographic measurement of possession trance in the field. AB - OBJECTIVES: To verify the utility of a portable electroencephalogram (EEG) measurement system developed for investigating spontaneous EEG from vigorously moving healthy subjects in a possession trance under a natural condition. METHODS: A portable multi-channel EEG telemetry system was developed to record the EEGs of 3 healthy male Balinese while they were performing a ritual dedicatory drama in the field. After reducing extraneous artifacts using a digital filter, the EEGs and their power spectra were analyzed in terms of evolution from one state to another. RESULTS: During the drama, one of the subjects became possessed while the others did not. The EEG of the possessed subject did not show any pathological findings including epileptic discharges, but indicated enhanced power in the theta and alpha frequency bands during the trance. This finding was not observed in the other two subjects, who did not go into trances, with no pathological EEG findings. CONCLUSIONS: The measurement system and data analysis methods we developed have allowed us, for the first time, to obtain an EEG from healthy subjects who are vigorously moving while in a possession trance. The present technique enables us to use a spontaneous EEG as a marker of the underlying physiology of a state of possession trance. PMID- 11897545 TI - Effects of intravenous mannitol on EEG recordings in stroke patients. AB - OBJECTIVES: To evaluate the usefulness of continuous EEG monitoring of stroke patients during and after intravenously infused mannitol. METHODS: Patients were rapidly administered 50 g of intravenous mannitol solution with continuous EEG monitoring for 3h pre- and post-drug infusion in the neurological intensive care unit. Visual and spectral analyses of EEG recording pre- and post-mannitol infusion were carried out. RESULTS: The study consisted of 47 patients. Of 38 patients with intracranial hemorrhage, 33 had abnormal EEG findings pre-mannitol administration. After mannitol therapy, visual analysis of the drug-induced EEG changes showed that the EEG findings were unchanged in 13 patients, demonstratively improved in 22 patients, and worse in 3 patients. The spectral analysis demonstrated that mannitol-induced EEG changes increased in alpha power and decreased in delta power in the lesion hemispheres, especially in the central and middle temporal areas. Maximal effects occurred 30 min post-mannitol infusion, and remained significant for 2h post-infusion. Of the 9 patients with cerebral infarction, only one with diffuse background slowing of one-side dominance pre-mannitol improved after the infusion of mannitol. CONCLUSIONS: The results of our investigation indicated that continuous EEG monitoring of mannitol treatment can reflect the brain edema, raised ICP in stroke patients, and provide assessment the drugs effects of antiedema and intracranial pressure lowering in vivo. PMID- 11897546 TI - Effects of intravenous human albumin and furosemide on EEG recordings in patients with intracerebral hemorrhage. AB - OBJECTIVES: To characterize alterations in continuous EEG monitoring that occurs during and after intravenous infusion of human albumin or furosemide in patients with intracerebral hemorrhage. METHODS: Patients were rapidly administered 20% human albumin 50 ml or furosemide 40 mg intravenously with continuous EEG monitoring for 3h before and after drug infusion in the neurological intensive care unit. Visual and spectral analyses of EEG recordings before and after mannitol administration were carried out. RESULTS: The study consisted of 20 patients. Of 14 patients with human albumin treatment, a decrease in the slowing activity was visually noted in 9 cases after the drug infusion. The spectral analysis demonstrated that albumin-induced EEG changes increased in alpha power and decreased in delta power in the lesion hemispheres, especially in the central and middle temporal areas. The effects occurred after 30 min and were maximal 1h after the end of the infusion, then remained significant for 2h post-infusion. Of 6 patients with furosemide treatment, the EEG recordings before, during, and after the furosemide infusion were not statistically significantly different by visual and quantitative analyses. CONCLUSIONS: The results support the opinion that the available EEG monitoring techniques offer an inexpensive, non-invasive, and consistently reproducible technique for reflecting the therapeutic effects of therapeutics in lowering ICP and antiedema in stroke patients. PMID- 11897547 TI - Botanical antioxidants for chemoprevention of photocarcinogenesis. AB - The incidence of non-melanoma skin cancer, consisting of basal- and squamous- cell carcinoma, continues to increase in the United States and elsewhere. Solar ultraviolet (UV) B radiation has been implicated as its main cause. This adverse effect of UVB has become a major human health concern. Therefore, development of novel strategies to reduce the occurrence of skin cancer is a highly desirable goal. Because UV radiation is known to cause excessive generation of reactive oxygen species (ROS) thereby resulting in an oxidative stress condition, the approaches aimed at counteracting ROS production may be useful for the prevention of skin cancer. One approach to reduce its occurrence is through 'Photochemoprotection', which we define as 'the use of agents capable of ameliorating the adverse effects of UVB on the skin'. Among many photochemoprotective agents, botanical antioxidants are showing promise. We propose that the use of botanical antioxidants, in combination with the use of sunscreens and educational efforts to avoid excessive sun exposure, may be an effective strategy for reduction of incidence of skin cancer and other UV mediated damage in humans. PMID- 11897548 TI - Pro-oxidant and anti-oxidant mechanism(s) of BHT and beta-carotene in photocarcinogenesis. AB - An hypothesis for the role of free radicals in cancer was elaborated by D. Harman in 1962 who suggested that it might be possible to reduce the extent of damage caused by free radicals through three dietary changes: (i) caloric reduction, i.e., lowering the level of free radical reactions arising in the course of normal metabolism; (ii) minimize dietary components that tend to increase the level of free radical reactions (e.g., polyunsaturated fats); and (iii) supplement the diet with one or more free radical reaction inhibitors (anti oxidants). With respect to (ii) and (iii), lipid peroxidation exemplifies the type of chain reaction initiated by free radicals, with unsaturated fatty acids being the primary center of free radical attack. Anti-oxidants act as free radical scavengers and are able to terminate these reactions. Indeed, the phenolic anti-oxidant butylated hydroxytoluene (BHT), and the carotenoid beta carotene, have both been shown to influence photocarcinogenesis, although the lack of correlation between physicochemical parameters and pathophysiological responses is apparent in both instances. The bimolecular rate constant for reaction of BHT with model peroxyl radicals is low while beta-carotene is highly reactive. However, both are able to efficiently inhibit lipid peroxidation reactions in biological membranes. Indeed, the influence of photocarcinogenesis by both BHT and beta-carotene is diminished as the level of dietary fat decreases, pointing to the involvement of lipid peroxidative reactions. Nevertheless, the mode of action of BHT in inhibiting photocarcinogenesis appears to be related to dose-diminution resulting from an increased spectral absorbance of the stratum corneum. On the other hand, beta-carotene has no such effect and may actually exacerbate photocarcinogenesis under certain dietary conditions. This paradox points to the complex relationship between chemical mechanisms and biological mode of action of anti-oxidants in photocarcinogenesis. Recent clinical and experimental data also suggest that supplementation of the complex and intricately balanced natural antioxidant defense system with one or more anti oxidants as a cancer prevention strategy will demand extreme caution. PMID- 11897549 TI - Postsynaptic calcium signaling microdomains in neurons. AB - Calcium ions are crucial messengers in the regulation of synaptic efficacy. In the postsynaptic neuron, this is exemplified by the tight temporal and spatial co segregation of calcium ions with calcium-dependent signal transduction protein complexes in dendritic spines. Over the last several years optical imaging, physiological, structural, and biological studies have clarified the molecular mechanisms underlying differential calcium signaling within the spine. In this review, we discuss how calcium signaling "microdomains" are organized and regulated. We emphasize the structural and functional features of precisely regulated supramolecular complexes incorporating proteins involved in calcium influx, calcium efflux, and signal transduction. These complexes act in concert to orchestrate the sophisticated postsynaptic calcium signaling that underlies synaptic plasticity. PMID- 11897550 TI - Peritoneal molecular environment, adhesion formation and clinical implication. AB - Whether induced by infection, inflammation, ischemia, and/or surgical injury, peritoneal adhesions are the leading cause of pelvic pain, bowel obstruction and infertility. It is clear that while postsurgical peritoneal wounds heal without adhesions in some patients, others develop severe scarring from seemingly equal procedures; in addition, in the same patient, adhesions can develop at one surgical site and not in another. The mechanisms underlying the predisposition to form adhesions as well as their site specificity are completely unknown. However, a large number of intraperitoneal surgical procedures are performed each day in the USA, and thus many patients are at risk of developing postoperative adhesions. Therefore, understanding of adhesion formation at the molecular level is essential and in the absence of such information, attempts to prevent patients from developing adhesions will remain an empirical process. The unprecedented advancement in molecular biology during the past decade has led to the identification of many biologically active molecules with the potential of regulating inflammatory and immune responses, angiogenesis and tissue remodeling, events that are central to normal peritoneal wound healing and adhesion formation. Although, the insight into their importance in the development of tissue fibrosis has substantially increased, their major roles in peritoneal biological functions and adhesion formation remain at best speculative. This article reviews the clinical implications of adhesions and attempts to highlight some of the key molecules i.e. growth factors, cytokines, chemokines, proteases and extracellular matrix, that are recognized to regulate inflammation, fibrinolysis, angiogenesis, and tissue remodeling, events that are central to peritoneal wound repair and adhesion formation. Finally, the article discusses the potential application and site specific delivery of several active compounds that are developed to alter the local inflammatory and immune response i.e., cytokine/chemokine network, targeted gene delivery and development of a new generation of biomaterials to prevent adhesion formation. Such understanding of peritoneal biology not only assist us to better manage patients with adhesion, but also those with endometriosis and malignant diseases that affect the peritoneal cavity. PMID- 11897551 TI - UV damage, DNA repair and skin carcinogenesis. AB - Skin cancer is unique among human cancers in its etiology, accessibility and the volume of detailed knowledge now assembled concerning its molecular mechanisms of origin. The major carcinogenic agent for most skin cancers is well established as solar ultraviolet light. This is absorbed in DNA with the formation of UV specific dipyrimidine photoproducts. These can be repaired by nucleotide excision repair or replicated by low fidelity class Y polymerases. Insufficient repair followed by errors in replication produce characteristic mutations in dipyrimidine sequences that may represent initiation events in carcinogenesis. Chronic exposure to UVB results in disruption of the epithelial structure and expansion of pre-malignant clones which undergo further genomic changes leading to full malignancy. Genetic diseases in DNA repair, xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy, show varied elevated symptoms of sun sensitivity involving skin cancers and other symptoms including neurological degeneration and developmental delays. In humans, only xeroderma pigmentosum shows high levels of cancer, but mouse strains, with any of the genes corresponding to these diseases knocked-out, show elevated skin carcinogenesis. The three major skin cancers exhibit characteristic molecular changes defined by certain genes and associated pathways. Squamous cell carcinoma involves mutations in the p53 gene; basal cell carcinoma involves mutations in the PATCHED gene, and melanoma in the p16 gene. The subsequent development of malignant tumors involves many additional genomic changes that have yet to be fully cataloged. PMID- 11897552 TI - Growth hormone and insulin-like growth factor-I in symptomatic and asymptomatic patients with diffuse idiopathic skeletal hyperostosis (DISH). AB - Basal serum growth hormone (GH) and insulin-like growth factor-I (IGF-I) concentration was measured by radioimmunoassay in patients with diffuse idiopathic skeletal hyperostosis (DISH) with muscle and joint pain and stiffness (symptomatic group) and in DISH patients without these constitutional clinical symptoms (asymptomatic group), but with persistent radiographic evidence of DISH. Serum GH and IGF-I was also measured in normal volunteers (control group) matched for gender and age to patients with DISH. Symptomatic male and female DISH patients had elevated serum GH and IGF-I concentration when compared to the control group. Asymptomatic DISH patients had serum GH levels that were significantly lower than their symptomatic counterparts. Clinical improvement did not alter serum IGF-I concentration. We conclude that serum GH concentration could be employed to monitor clinical remission in DISH. PMID- 11897553 TI - Oxidative processes in the brain and non-neuronal tissues as biomarkers of Alzheimer's disease. AB - Diminished metabolism and excessive oxidative stress occur in the brains of patients with Alzheimer's Disease (AD). These abnormalities in oxidative processes occur in the brain in early stages of AD, which suggests that the deficits are not just secondary to the neuro-degeneration. Alterations in oxidative processes also occur in early stages of AD in non-neuronal tissues including fluids (e.g., cerebrospinal fluid, plasma and urine), cell like particles (e.g., red blood cells and platelets) and cells (e.g., lymphocytes). AD related abnormalities also persist in cultured cells such as fibroblasts, which indicates that the AD-related changes are not secondary to pathology, and reflect inherent properties of AD cells. These measures of abnormalities in oxidative processes in peripheral cells from AD patients have the potential to be useful as diagnostic markers, as indicators of the progression of the disease, as a tool to develop therapeutic approaches and as monitors of therapeutic efficacy. The peripheral cells are also useful for discovering mechanisms that underlie the multiple changes in cell signaling pathways that accompany AD. Several experimental approaches suggest that oxidative stress is a convergence factor that leads to many other AD-related changes. This review focuses on the considerable recent progress in the quest for markers of metabolism/oxidative stress in peripheral tissues from AD patients, and on experiments to test their pathophysiological importance. PMID- 11897554 TI - Transition metal chelator therapy--a potential treatment for Alzheimer's disease? AB - A defining feature of Alzheimer's disease (AD) pathology is the presence of amyloid beta known as A-beta (Abeta) within neuritic plaques of the hippocampus and neocortex of the brain. While early in vitro studies suggested that Abeta could itself be toxic to neuronal cells, recent studies have indicated that this peptide has both neurotoxic and neuroprotective properties that are modulated by the binding of transition metal ions. Transition metal ion binding was shown to modulate Abeta solubility as well as its hydrogen peroxide production, thereby providing explanations for both its trophic and toxic properties. These findings lead to the suggestion that interference with this interaction may reverse the neurotoxic properties of Abeta. More recently, in vivo and in vitro studies into the effects of transition metal chelator treatments on Abeta solubilisation and neurological function have been published. Such studies have yielded promising results, however the potential side effects of many such metal chelators may prove too great for clinical use. It is widely agreed that the ideal chelator for such interdiction would act only on those transition metals that complex with Abeta, and only at metal ion binding sites that contribute to Abeta aggregation and reactive oxygen species generation. The efficacy of metal chelators in reducing Abeta load in transgenic mouse brains demonstrates that this approach has considerable merit as a research tool and as a stimulus to develop second generation agents that can selectively prevent transition metals from binding to the Abeta peptide itself without perturbing the action of other important metal requiring biomolecules in the brain. PMID- 11897555 TI - Roles of TGF-beta in hepatic fibrosis. AB - TGF-beta has multiple profibrogenic but also anti-inflammatory and immunosuppressive effects. The balance of these actions is required for maintaining tissue homeostasis and an aberrant expression of TGF-beta is involved in a number of disease processes in the liver. In addition to its fibrogenic action leading to transdifferentiation of hepatic stellate cells into myofibroblasts, TGF-beta is also an important negative regulator of proliferation and an inducer of apoptosis. The major portion of TGF-beta is secreted as part of an inactive complex and the details of the activation process in liver have not yet been elucidated. The initially striking simplicity of the core TGF-beta/Smad signaling pathways is rapidly giving way to a much more complex view of intracellular signal transduction mechanisms and recent work has demonstrated the importance of cross-talk among different signaling pathways to either specify, enhance, or inhibit TGF-beta responses. The ubiquitous pathophysiologic relevance of TGF-beta suggests its measurement in blood as a diagnostic tool. Other strategies aim at inhibition of TGF-beta1 function or synthesis as a primary target for the development of antifibrotic strategies and recent advances in cell biology have opened several ways to approach the inhibition of TGF-beta action. PMID- 11897556 TI - Pi 3-kinase and its up- and down-stream modulators as potential targets for the treatment of type II diabetes. AB - Type 2 diabetes is caused by a combination of impaired insulin secretion and, to a greater extent, resistance of target tissues to insulin action. Phosphoinositide 3-kinase (PI3K) plays a key role in insulin signaling and has been shown to be blunted in tissues of type 2 diabetes subjects. There is emerging biochemical and, particularly, genetic evidence suggesting that insulin resistance can potentially be treated via modulation of PI3K by targeting PI3K itself or its up and down-stream modulators. These potential targets include Src homology 2 domain containing inositol 5-phosphatase 2 (SHIP2), phosphatase and tensin homolog deleted on chromosome ten (PTEN), kappaB kinase beta (IKKbeta), PKC isoforms, and the PI3K p85 subunit. There is evidence suggesting that their inhibition affects PI3K activity and improves insulin sensitivity in vivo. In the current review, we will discuss the role of these molecules in insulin-mediated activation of PI3K, the rational for targeting these molecules for diabetes treatment, and some critical issues in terms of drug development. PMID- 11897557 TI - Voltage-sensor control of Ca2+ release in skeletal muscle: insights from skinned fibers. AB - Important aspects of the excitation-contraction (EC) coupling process in skeletal muscle have been revealed using mechanically-skinned fibers in which the transverse-tubular system can be depolarized by ion substitution or electrical stimulation, activating the voltage-sensors which in turn open the Ca2+ release channels in the adjacent sarcoplasmic reticulum (SR). Twitch and tetanic force responses elicited in skinned fibers closely resemble those in intact fibers, showing that the coupling mechanism is entirely functional. It was found that ATP has to be bound to the Ca2+ release channels for them to be activated by the voltage-sensors and that the coupling mechanism likely involves the voltage sensors removing the inhibitory effects of cytoplasmic Mg2+ on the release channels; such findings are relevant to the basis of muscle fatigue and to certain diseases such as malignant hyperthermia (MH). EC coupling is evidently not mediated by upmodulation of Ca2+-induced Ca2+ release (CICR) or by an oxidation or phosphorylation reaction. The Ca2+ load in the SR of skinned fibers can be set at the endogenous level or otherwise. The normal coupling mechanism functions well in mammalian fast-twitch fibers even when the SR is only partially loaded, whereas CICR is highly dependent on SR luminal Ca2+ and caffeine is poorly effective at inducing release at the endogenous SR Ca2+ load level. PMID- 11897558 TI - Ryanodine receptors, FKBP12, and heart failure. AB - RyR2 function is regulated by highly conserved signaling pathways that modulate excitation-contraction (EC) coupling. cAMP dependent protein kinase (PKA) phosphorylation of RyR2 plays an important role in regulating channel function in response to stress signaled by the sympathetic nervous system (the classic "fight or flight response") (1). PKA phosphorylation of RyR2 induces dissociation of the regulatory protein FKBP12.6 resulting in channels with increased sensitivity to Ca2+-induced Ca2+ release. Under normal physiological conditions (no cardiac damage) PKA phosphorylation of RyR2 is part of an integrated physiological response that leads to increased EC coupling gain and increased cardiac output. PKA-hyperphosphorylation of RyR2 in failing hearts is a maladaptive response that results in depletion of FKBP12.6 from the RyR2 macromolecular complex and defective channel function (pathologically increased sensitivity to Ca2+-induced Ca2+ release) that may cause depletion of SR Ca2+ and diastolic release of SR Ca2+ that can initiate delayed after depolarizations (DADs) that trigger ventricular arrhythmias (1). RyR2 mutations in patients with catecholaminergic induced sudden cardiac death provide further evidence linking the sympathetic nervous system, RyR2 and ventricular arrhythmias (2-4). The chronic hyperadrenergic state of heart failure is associated with defective Ca2+ signaling in part due to PKA hyperphosphorylation of RyR2. PMID- 11897559 TI - Molecular mechanisms of photocarcinogenesis. AB - Photocarcinogenesis represents the accumulation of genetic changes as well as immune system modulation, which ultimately lead to the development of skin cancers. The recent advances in molecular and cellular biology have clarified the mechanisms of photocarcinogenesis, including the formation of DNA photoproducts, DNA repair, mutation of proto-oncogenes and tumor suppressor genes, and UV induced immunosuppression. The understanding and further investigation of photocarcinogenesis is critical to the development of effective prevention and intervention strategies for human skin cancer. PMID- 11897560 TI - Applications of enzymatic amplification staining in immunophenotyping hematopoietic cells. AB - Immunofluorescent staining of mammalian cells has provided a reliable approach for detection of specific antigen expression in situ. An advantage of fluorescent markers has been their applicability to automated, high-throughput cellular analysis by flow cytometry. Flow cytometry has thus become an integral component of clinical laboratory diagnostics, particularly in the areas of immunology and hematology. One of the major drawbacks of traditional immunofluorescent staining, even with flow cytometric detection, has been the difficulty in detecting low abundance cellular antigens, some of which may have clinical and scientific significance. To address these problems, staining techniques have recently been developed to increase the sensitivity of cellular antigen detection by flow cytometry. In this review we will describe a few of these techniques and focus on enzymatic amplification staining as a means to generate a highly augmented antigen-specific signal. We will also discuss practical applications of enzymatic amplification for immunostaining of clinical specimens. PMID- 11897561 TI - The AD7C-NTP neuronal thread protein biomarker for detecting Alzheimer's disease AB - Dementia in Alzheimer's disease (AD) is ultimately due to cell loss mediated by several mechanisms including, apoptosis, impaired mitochondrial function, and possibly necrosis. A second major neuroanatomic correlate of dementia is aberrant cortical neuritic sprouting with abundant proliferation of dystrophic neurites. Early in vivo detection of AD will require non-invasive assays of highly sensitive and relatively specific biomarkers that reflect these fundamental abnormalities in cellular function. The AD-associated neuronal thread protein (AD7c-NTP) gene encodes an approximately 41 kD membrane-spanning phosphoprotein that causes apoptosis and neuritic sprouting in transfected neuronal cells. The AD7c-NTP gene is over-expressed in AD beginning early in the course of disease. In the brain, increased AD7c-NTP immunoreactivity is associated with phospho-tau immunoreactive cytoskeletal lesions, but not with amyloid-? accumulations. The levels of AD7c-NTP in postmortem brain tissue correlate with the levels measured in paired ventricular fluid samples, suggesting that the protein is secreted or released by dying cells into cerebrospinal fluid (CSF). In this regard, elevated levels of AD7c-NTP can be detected in both CSF and urine of patients with early or moderately severe AD, and the CSF and urinary levels of AD7c-NTP correlate with the severity of dementia. The newest configuration of the AD7c-NTP assay, termed "7c Gold", has greater than 90% sensitivity and specificity for detecting early AD. The aggregate results from a number of studies suggest that AD7c-NTP is an excellent biomarker that could be helpful in the routine clinical evaluation of elderly patients at risk for AD. PMID- 11897562 TI - Protein tyrosine phosphorylation in T cell signaling. AB - This review discusses in considerable detail the tyrosine phosphorylation events that couple the T cell antigen receptor to downstream signaling pathways. First, the protein kinases that catalyze these tyrosine phosphorylation are introduced, then the phosphatases that mediate removal of phosphate from tyrosine residues. Next, we discuss the molecular clock work by which these enzymes initiated T cell activation. Finally, we briefly review the key substrates for tyrosine phosphorylation and their role in coupling the kinases and phosphatases to gene transcription and other aspects of the T lymphocyte response to antigen. PMID- 11897563 TI - Control of myeloid dendritic cell differentiation and function by CD1d-restricted (NK) T cells. AB - While regulating a wide variety of immunologic responses, the precise immunologic functions of CD1d-restricted (NK) T cells are not well defined. Notably, In vitro activation of human NK T cell clones results in the secretion of multiple cytokines important for the recruitment and differentiation of myeloid dendritic cells (DC). Once differentiated, these DC strongly activate NK T cells. In humans, CD1d is expressed by myeloid DC and on tumor cells of this lineage. Another specialized myeloid antigen presenting cell, the epithelioid histiocyte seen in granulomatous inflammation, also expresses CD1d. Because myeloid DC are important regulators of Th1/Th2 T cell responses, cross talk between human NK T cells and myeloid DC would be expected to have significant impact on many immune responses. Consistent with this hypothesis, NK T cells are required for myeloid DC-controlled antitumor responses in mice, and regulate diabetes in nonobese diabetic (NOD) mouse by locally controlling the frequency and function of DC subsets. Thus, regulation of myeloid DC by NK T cells controls both the transition from innate to adaptive immunity and the Th-phenotype of subsequent T cell responses. PMID- 11897564 TI - Activation of hepatic stellate cells--a key issue in liver fibrosis. AB - Hepatic fibrosis describes the presence of excess collagen due to new fiber formation, laid down as part of the tissue repair response to chronic liver injury. The causes of injury include toxins, disorders of the immune system, viral and parasitic infections, as well as rarer liver diseases such as haemochromatosis, Wilson's disease and galactosaemia. Whatever the cause of injury, the cells and soluble factors contributing to this wound healing response are similar. The principal effector of hepatic fibrogenesis is now widely recognized as the hepatic stellate cell. Stellate cells are usually quiescent cells, but in response to liver injury they undergo an activation process in which they become highly proliferative and synthesize a fibrotic matrix rich in type I collagen. Initiation of stellate cell activation is largely due to paracrine stimulation, whereas perpetuation of activation involves autocrine as well as paracrine loops, and is dependent on a number of functional changes. The principal paracrine and autocrine factors currently thought to be involved in these processes are discussed in this review, as are the roles of the extracellular matrix, the nuclear receptor superfamily, non-peptide ligands, and oxidative stress. PMID- 11897565 TI - Cerebrospinal fluid Abeta40 and Abeta42: natural course and clinical usefulness. AB - Amyloid beta protein 40 (Abeta40) and 42 Abeta42, major components of senile plaque amyloids, are physiological peptides present in the brain, cerebrospinal fluid (CSF) and plasma. The levels of CSF Abeta40 and Abeta42(43) show a U-shaped natural course in normal aging. The increase of Abeta42(43) over 60 years of age is inhibited in Alzheimer's disease (AD). This specific alteration of CSF Abeta42(43) correlates with Abeta deposits in the AD brain providing a biological basis for a biomarker of AD. In the GTT2 study, assays of the CSF Abeta ratio ((Abeta40/ Abeta42(43)) showed a diagnostic sensitivity (59%) and specificity (88%) compared with non-AD type dementia and controls. The levels of the Abeta ratio increased from early to late stages of AD. Combination assays of CSF tau and Abeta ratio provided further efficient diagnostic sensitivity (81%) and specificity (87%). The reliability of the assay may prompt worldwide usage of these CSF biomarkers for Alzheimer's patients. PMID- 11897566 TI - Prion peptide 106-126 as a model for prion replication and neurotoxicity. AB - Prion diseases or transmissible spongiform encephalopathies are neurodegenerative disorders that are genetic, sporadic, or infectious. The pathogenetic event common to all prion disorders is a change in conformation of the cellular prion protein (PrPC) to the scrapie isoform (PrPSc), which, unlike PrPC, aggregates easily and is partially resistant to protease digestion. Although PrPSc is believed to be essential for the pathogenesis and transmission of prion disorders, the mechanism by which PrPSc deposits cause neurodegeneration is unclear. It has been proposed that in some cases of prion disorders, a transmembrane form of PrP, termed CtmPrP may be the mediator of neurodegenerative changes rather than PrPSc per se. In order to understand the underlying cellular processes by which PrPSc mediates neurodegeneration, we have investigated the mechanism of neurotoxicity by a beta-sheet rich peptide of PrP in a cell model. We show that exposure of human neuronal cell lines NT-2 and M17 to the prion peptide 106-126 (PrP106-126) catalyzes the aggregation of endogenous cellular prion protein (PrPC) to an amyloidogenic form that shares several characteristics with PrPSc. Intracellular accumulation of these PrPSc-like forms upregulates the synthesis of CtmPrP, which is proteolytically cleaved in the endoplasmic reticulum and the truncated C-terminal fragment is transported to the cell surface. In addition, we have isolated mutant NT-2 and neuroblastoma cells that are resistant to toxicity by PrP106-126 to facilitate further characterization of the biochemical pathways of PrP106-126 neurotoxicity. The PrP106-126-resistant phenotype of these cells could result from aberrant binding or internalization of the peptide, or due to an abnormality in the downstream pathway(s) of neuronal toxicity. Thus, our data suggest that PrPSc aggregation occurs by a process of 'nucleation' on a pre-existing 'seed' of PrP. Furthermore, the PrP106-126 resistant cells reported here will provide a unique opportunity for identifying the cellular and biochemical pathways that mediate neurotoxicity by PrPSc. PMID- 11897567 TI - Alzheimer's disease biomarkers: their value in diagnosis and clinical trials. AB - Biomarkers of Alzheimer's disease may be useful, not only for early diagnosis of the disease, but also for monitoring the progress of drug trials. A number of plasma and cerebrospinal fluid markers are reportedly altered in Alzheimer's disease. So far, no single biomarker can be used to diagnose Alzheimer's disease definitively. Nevertheless, it may eventually be possible to use several markers in combination to obtain sufficient diagnostic accuracy. However, for this to be the case, new specific biomarkers may need to be identified. PMID- 11897568 TI - 3'-phosphoinositide-dependent kinase-1 (PDK-1) in PI 3-kinase signaling. AB - The recently discovered 3'-phosphoinositide-dependent kinase-1 (PDK-1) is a serine/threonine protein kinase which phosphorylates several members of the conserved AGC kinase superfamily (comprising the prototypes protein kinases A (PKA), G (PKG) and C (PKC)). Phosphorylation of a threonine or serine residue in the activation loop (also known as the T-loop) of these kinases is a critical step in their activation, and is typically accompanied by additional phosphorylations elsewhere in the molecule. Phosphorylation of the activation loop is a common regulatory mechanism shared by most serine/threonine as well as tyrosine kinases as it facilitates alignment of amino acid residues in the active site which are involved in the phosphotransferase reaction. Therefore the discovery of PDK-1 as the enzyme which mediates this event in many protein kinases introduced a new and important step in signaling pathways which regulate numerous important cellular processes including cellular survival, glucose transport and metabolism, tumor progression as well as protein translation. Moreover, the finding that PDK-1 function is mediated in part by the phosphoinositide 3'-OH-kinase (PI 3-K) pathway also provided an explanation as to how the lipid products of PI 3-K, namely phosphatidylinositol-3,4-bisphosphate (PtdIns-3,4-P2) and phosphatidylinositol-3,4-5-trisphosphate (PtdIns-3,4,5-P3) stimulate the activation of protein kinase-dependent signaling pathways. These initial landmark observations were followed by many important studies which provided additional mechanistic insight into both PDK-1 regulation as well as the role of this kinase in cellular function. This review will focus on the regulation of PDK-1 and the various mechanisms which it uses to contribute to the activation of target kinases. PMID- 11897569 TI - Significance of intracellular Abeta42 accumulation in Alzheimer's disease. AB - Abeta plays a pivotal role in the pathogenesis of Alzheimer's disease (AD), but it is still obscure how it causes AD. We have established transgenic mice carrying wild-type or familial Alzheimer's disease (FAD) mutant-type presenilin 1 (PS1). In these mice, the number of cortical and hippocampal neurons decreased along with age in mutant mice. In addition, the old mutant mice showed a significant increase of dark neurons by silver staining and the number of neurons with intracellular Abeta42 by immunohistochemistry. Our extended study also showed a significant increase of intracellular Abeta42-positive neurons in isolated cases of AD as well as in PS1 mutant FAD cases. These neurons frequently showed apoptotic staining. However, coincidence of apoptotic markers and intraneuronal neurofibrillary tangles (NFT) was insignificant. Notably intraneuronal Abeta42-labeling was frequently seen in a case of AD showing cotton wool type senile plaques with a few NFT positive neurons and dystrophic neurites. These results indicate that intraneuronal deposition of Abeta42 is important in the pathogenesis of AD. PMID- 11897570 TI - Integrative analysis of calcium signalling in cardiac muscle. AB - This review discusses the control of the amplitude of the cardiac systolic Ca transient. The Ca transient arises largely from release from the sarcoplasmic reticulum (SR). Release is triggered by calcium-induced calcium release (CICR) whereby the entry of a small amount of Ca on the L-type Ca current, "the trigger", results in the release of much more Ca from the SR. There are three potential control points: (1) the Ca content of the SR; (2) the properties of the SR Ca release channel or ryanodine receptor (RyR); (3) the amplitude of the L type Ca current. The data reviewed show that the Ca content of the SR has pronounced effects on systolic [Ca2+]i and, reciprocally, the amount of Ca released from the SR affects sarcolemmal Ca fluxes thereby "autoregulating" SR content. Modulation of the ryanodine receptor has no steady-state effect due to compensating changes of SR Ca content. An increase of the L-type Ca current results in an abrupt increase of systolic [Ca2+]i with little change of SR content. This is because of a coordinated increase of both the trigger and loading function of the Ca current. These results emphasise the importance of considering all aspects of Ca handling in the context of SR Ca release and thus the regulation of the systolic Ca transient and contraction in cardiac muscle. PMID- 11897571 TI - The zebrafish as a model system for human disease. AB - The zebrafish (Danio rerio) has been widely utilised for the study of developmental biology, which has lead to the evolution of sophisticated cellular and molecular approaches. More recently, the rapid progress of various zebrafish genomic infrastructure initiatives is facilitating the development of zebrafish models of human disease. This review aims to describe several representative examples of how the zebrafish can be successfully used to identify novel genes and assign gene function, providing invaluable clues to human pathophysiology. PMID- 11897572 TI - Immunotherapy of human papillomavirus-associated malignancies and the challenges posed by T-cell tolerance. AB - Human papillomaviruses are associated with a broad range of carcinomas, including cervical cancer. Although the delivery of immunogenic tumor-associated antigens represents a promising approach in the treatment of these malignancies, the imposition of T cell tolerance poses a significant challenge in this endeavor. The purpose of this review is to discuss T cell tolerance and the role of T cell costimulation in the immunotherapy of HPV-associated malignancies. PMID- 11897573 TI - Development of a whole-cell assay for peptidoglycan biosynthesis inhibitors. AB - Osmotically stabilized Escherichia coli cells subjected to freezing and thawing were utilized as the source of enzymes for a peptidoglycan pathway assay that can be used to simultaneously test all targets of the committed steps of cell wall biosynthesis. The use of (14)C-labeled UDP-N-acetylglucosamine (UDP-GlcNAc) as a substrate allows the direct detection of cross-linked peptidoglycan formed. The assay was validated with known antibiotics. Fosfomycin was the strongest inhibitor of the pathway assay, with a 50% inhibitory concentration of 1 microM. Flavomycin, bacitracin, vancomycin, D-cycloserine, penicillin G, and ampicillin also inhibited formation of radiolabeled peptidoglycan by the E. coli cells. Screening of compounds identified two inhibitors of the pathway, Cpd1 and Cpd2. Subsequent tests with a biochemical assay utilizing purified enzyme implicated UDP-GlcNAc enolpyruvyl transferase (MurA) as the target of Cpd1. This compound inhibits the first enzyme of the pathway in a time-dependent manner. Moreover, enzyme inactivation is dependent on preincubation in the presence of UDP-GlcNAc, which forms a complex with MurA, exposing its active site. Cpd1 also displayed antimicrobial activity against a panel of microorganisms. The pathway assay used in conjunction with assays for individual enzymes provides an efficient means of detecting and characterizing novel antimicrobial agents. PMID- 11897574 TI - Candida albicans sterol C-14 reductase, encoded by the ERG24 gene, as a potential antifungal target site. AB - The incidence of fungal infections has increased dramatically, which has necessitated additional and prolonged use of the available antifungal agents. Increased resistance to the commonly used antifungal agents, primarily the azoles, has been reported, thus necessitating the discovery and development of compounds that would be effective against the major human fungal pathogens. The sterol biosynthetic pathway has proved to be a fertile area for antifungal development, and steps which might provide good targets for novel antifungal development remain. The sterol C-14 reductase, encoded by the ERG24 gene, could be an effective target for drug development since the morpholine antifungals, inhibitors of Erg24p, have been successful in agricultural applications. The ERG24 gene of Candida albicans has been isolated by complementation of a Saccharomyces cerevisiae erg24 mutant. Both copies of the C. albicans ERG24 gene have been disrupted by using short homologous regions of the ERG24 gene flanking a selectable marker. Unlike S. cerevisiae, the C. albicans ERG24 gene was not required for growth, but erg24 mutants showed several altered phenotypes. They were demonstrated to be slowly growing, with doubling times at least twice that of the wild type. They were also shown to be significantly more sensitive to an allylamine antifungal and to selected cellular inhibitors including cycloheximide, cerulenin, fluphenazine, and brefeldin A. The erg24 mutants were also slightly resistant to the azoles. Most importantly, erg24 mutants were shown to be significantly less pathogenic in a mouse model system and failed to produce germ tubes upon incubation in human serum. On the basis of these characteristics, inhibitors of Erg24p would be effective against C. albicans. PMID- 11897575 TI - Malaria parasites giving rise to recrudescence in vitro. AB - Recrudescences were simulated in vitro with drug treatment to examine how drug sensitive parasites survive the treatment. Various numbers of cultured parasites were treated with lethal doses of pyrimethamine or mefloquine for various lengths of time. Recrudescences were observed in parasite populations with larger initial numbers of parasites when the treatment duration was prolonged. Equal numbers of parasitized erythrocytes were treated with various concentrations of pyrimethamine or mefloquine. There was no clear linear relationship between the incidence of recrudescence and the drug concentration. Parasites that had recrudesced were continuously allowed to recrudesce in the succeeding recrudescence experiments. Both the duration from the cessation of treatment to the time at which the recrudescent parasitemia level reached 1% and the growth rate of recrudescent parasites were equal among these recrudescences. The recrudescent parasites in these experiments were as sensitive to the drugs as the parasites tested before treatment were. These results suggest that a parasite culture may contain parasites in some phases that are not killed by drug for up to 10 days, which explains the recrudescences that occur even after treatment. PMID- 11897576 TI - Molecular and biochemical characterization of Ambler class A extended-spectrum beta-lactamase CGA-1 from Chryseobacterium gleum. AB - Antibiotic susceptibility testing by disk diffusion of a Chryseobacterium gleum isolate, strain CIP 103039, showed a typical synergy image between clavulanic acid and expanded-spectrum cephalosporins. Shotgun cloning gave a recombinant plasmid in Escherichia coli that produced a beta-lactamase, CGA-1, with a pI value of 8.9 that conferred resistance to most penicillins (except ureidopenicillins) and narrow-spectrum cephalosporins and an intermediate susceptibility to expanded-spectrum cephalosporins and aztreonam. The CGA-1 amino acid sequence shared only 60% amino acid identity with CME-1 and CME-2 from Chryseobacterium meningosepticum, the most closely related beta-lactamases. CGA-1 was very likely chromosome encoded. It is a novel member of the PER subgroup of Ambler class A beta-lactamases (Bush functional group 2be). PMID- 11897578 TI - In vitro and in vivo activities of anti-influenza virus compound T-705. AB - T-705 (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) has been found to have potent and selective inhibitory activity against influenza virus. In an in vitro plaque reduction assay, T-705 showed potent inhibitory activity against influenza A, B, and C viruses, with 50% inhibitory concentrations (IC(50)s) of 0.013 to 0.48 microg/ml, while it showed no cytotoxicity at concentrations up to 1,000 microg/ml in Madin-Darby canine kidney cells. The selectivity index for influenza virus was more than 2,000. It was also active against a neuraminidase inhibitor resistant virus and some amantadine-resistant viruses. T-705 showed weak activity against non-influenza virus RNA viruses, with the IC(50)s being higher for non influenza virus RNA viruses than for influenza virus, and it had no activity against DNA viruses. Orally administered T-705 at 100 mg/kg of body weight/day (four times a day) for 5 days significantly reduced the mean pulmonary virus yields and the rate of mortality in mice infected with influenza virus A/PR/8/34 (3 x 10(2) PFU). These results suggest that T-705 may be a compound that is useful and highly selective against influenza virus infections and that has a mode of action different from those of commercially available drugs, such as amantadine, rimantadine, and neuraminidase inhibitors. PMID- 11897577 TI - In vitro and in vivo activities of a novel cephalosporin, BMS-247243, against methicillin-resistant and -susceptible staphylococci. AB - The recent emergence of methicillin-resistant Staphylococcus aureus (MRSA) with decreased susceptibility to vancomycin has intensified the search for alternative therapies for the treatment of infections caused by this organism. One approach has been to identify a beta-lactam with improved affinity for PBP 2a, the target enzyme responsible for methicillin resistance in staphylococci. BMS-247243 is such a candidate, with MICs that inhibit 90% of isolates tested (MIC(90)s) of 4, 2, and 8 microg/ml for methicillin-resistant strains of S. aureus, S. epidermidis, and S. haemolyticus, respectively, as determined on plates with Mueller-Hinton agar and 2% NaCl. The BMS-247243 MICs for MRSA were minimally affected by the susceptibility testing conditions (inoculum size, prolonged incubation, addition of salt to the test medium) or by staphylococcal beta lactamases. BMS-247243 MIC(90)s for methicillin-susceptible staphylococcal species ranged from < or = 0.25 to 1 microg/ml. The BMS-247243 MIC(90) for beta lactamase-producing S. aureus strains was fourfold higher than that for beta lactamase-nonproducing strains. BMS-247243 is hydrolyzed by staphylococcal beta lactamases at 4.5 to 26.2% of the rates measured for cephaloridine. The affinity of BMS-247243 for PBP 2a was >100-fold better than that of methicillin or cefotaxime. BMS-247243 is bactericidal for MRSA, killing the bacteria twice as fast as vancomycin. These in vitro activities of BMS-247243 correlated with its in vivo efficacy against infections in animals, including the neutropenic murine thigh and rabbit endocarditis models involving MRSA strains. In conclusion, BMS 247243 has in vitro and in vivo activities against methicillin-resistant staphylococci and thus may prove to be useful in the treatment of infections caused by these multidrug-resistant organisms. PMID- 11897579 TI - Hemofiltrate CC chemokine 1[9-74] causes effective internalization of CCR5 and is a potent inhibitor of R5-tropic human immunodeficiency virus type 1 strains in primary T cells and macrophages. AB - Proteolytic processing of the abundant plasmatic human CC chemokine 1 (HCC-1) generates a truncated form, HCC-1[9-74], which is a potent agonist of CCR1, CCR3, and CCR5; promotes calcium influx and chemotaxis of T lymphoblasts, monocytes, and eosinophils; and inhibits infection by CCR5-tropic human immunodeficiency virus type 1 (HIV-1) isolates. In the present study we demonstrate that HCC-1[9 74] interacts with the second external loop of CCR5 and inhibits replication of CCR5-tropic HIV-1 strains in both primary T cells and monocyte-derived macrophages. Low concentrations of the chemokine, however, frequently enhanced the replication of CCR5-tropic HIV-1 isolates but not the replication of X4 tropic HIV-1 isolates. Only HCC-1[9-74] and HCC-1[10-74], but not other HCC-1 length variants, displayed potent anti-HIV-1 activities. Fluorescence-activated cell sorter analysis revealed that HCC-1[9-74] caused up to 75% down-regulation of CCR5 cell surface expression, whereas RANTES (regulated on activation, normal T-cell expressed and secreted) achieved a reduction of only about 40%. Studies performed with green fluorescent protein-tagged CCR5 confirmed that both HCC-1[9 74] and RANTES, but not full-length HCC-1, mediated specific internalization of the CCR5 HIV-1 entry cofactor. Our results demonstrate that the interaction with HCC-1[9-74] causes effective intracellular sequestration of CCR5, but they also indicate that the effect of HCC-1[9-74] on viral replication is subject to marked cell donor- and HIV-1 isolate-dependent variations. PMID- 11897580 TI - Enhanced inhibition of orthopoxvirus replication in vitro by alkoxyalkyl esters of cidofovir and cyclic cidofovir. AB - The nucleotide phosphonates cidofovir (CDV) and cyclic cidofovir (cCDV) are potent antiviral compounds when administered parenterally but are not well absorbed orally. These compounds have been reported to have activity against orthopoxvirus replication in vitro and in animal models when administered parenterally or by aerosol. To obtain better oral activity, we synthesized a novel series of analogs of CDV and cCDV by esterification with two long-chain alkoxyalkanols, 3-hexadecyloxy-1-propanol (HDP-CDV; HDP-cCDV) or 3-octadecyloxy-1 ethanol (ODE-CDV; ODE-cCDV). Their activities were evaluated and compared with those of CDV and cCDV in human foreskin fibroblast (HFF) cells infected with vaccinia virus (VV) or cowpox virus (CV) using a plaque reduction assay. The 50% effective concentrations (EC(50)s) against VV in HFF cells for CDV and cCDV were 46.2 and 50.6 microM compared with 0.84 and 3.8 microM for HDP-CDV and HDP-cCDV, respectively. The EC(50)s for ODE-CDV and ODE-cCDV were 0.20 and 1.1 microM, respectively. The HDP analogs were 57- and 13-fold more active than the parent nucleotides, whereas the ODE analogs were 231- and 46-fold more active than the unmodified CDV and cCDV. Similar results were obtained using CV. Cytotoxicity studies indicated that although the analogs were more toxic than the parent nucleotides, the selective index was increased by 4- to 13-fold. These results indicate that the alkoxyalkyl esters of CDV and cCDV have enhanced activity in vitro and need to be evaluated for their oral absorption and efficacy in animal models. PMID- 11897581 TI - Oral administration of cyclopentane neuraminidase inhibitors protects ferrets against influenza virus infection. AB - Several cyclopentane inhibitors of influenza virus neuraminidase that have inhibitory activities in tissue culture similar to those of zanamivir and oseltamivir have recently been described. These new inhibitors have been examined for efficacy against a virulent H3N2 influenza virus when administered orally to infected ferrets. Preliminary studies indicated that oral administration of BCX 1923, BCX-1827, or BCX-1812 (RWJ-270201) at a dose of 5 or 25 mg/kg of body weight was active in ferrets in reducing respiratory and constitutional signs and symptoms, but these antivirals affected virus titers in the upper and lower respiratory tracts only marginally. Of the three compounds, BCX-1812 seemed to be the most efficacious and was examined further at higher doses of 30 and 100 mg/kg. These doses significantly reduced peak virus titers in nasal washes and total virus shedding as measured by areas under the curve. Virus titers in lung homogenates were also reduced compared to those in controls, but the difference was not statistically significant. As was observed with BCX-1812 at lower doses, the nasal inflammatory cellular response, fever, and nasal signs were reduced while ferret activity was not, with activity remaining similar to uninfected animals. PMID- 11897583 TI - In vitro characterization of A-315675, a highly potent inhibitor of A and B strain influenza virus neuraminidases and influenza virus replication. AB - A-315675 is a novel, pyrrolidine-based compound that was evaluated in this study for its ability to inhibit A and B strain influenza virus neuraminidases in enzyme assays and influenza virus replication in cell culture. A-315675 effectively inhibited influenza A N1, N2, and N9 and B strain neuraminidases with inhibitor constant (K(i)) values between 0.024 and 0.31 nM. These values were comparable to or lower than the K(i) values measured for oseltamivir carboxylate (GS4071), zanamivir, and BCX-1812, except for the N1 enzymes that were found to be the most sensitive to BCX-1812. The time-dependent inhibition of neuraminidase catalytic activity observed with A-315675 is likely due to its very low rate of dissociation from the active site of neuraminidase. The half times for dissociation of A-315675 from B/Memphis/3/89 and A/Tokyo/3/67 (H3N2) influenza virus neuraminidases of 10 to 12 h are significantly slower than the half times measured for oseltamivir carboxylate (33 to 60 min). A-315675 inhibited the replication of several laboratory strains of influenza virus in cell culture with potencies that were comparable or superior to those for oseltamivir carboxylate and BCX-1812, except for the A/H1N1 viruses that were found to be two- to fourfold more susceptible to BCX-1812. A-315675 and oseltamivir carboxylate exhibited comparable potencies against a panel of A/H1N1 and A/H3N2 influenza virus clinical isolates, but A-315675 was found to be significantly more potent than oseltamivir carboxylate against the B strain isolates. The favorable in vitro results relative to other clinically effective agents provide strong support for the further investigation of A-315675 as a potential therapy for influenza virus infections. PMID- 11897582 TI - Kinetic analysis of wild-type and YMDD mutant hepatitis B virus polymerases and effects of deoxyribonucleotide concentrations on polymerase activity. AB - Mutations in the YMDD motif of the hepatitis B virus (HBV) DNA polymerase result in reduced susceptibility of HBV to inhibition by lamivudine, at a cost in replication fitness. The mechanisms underlying the effects of YMDD mutations on replication fitness were investigated using both a cell-based viral replication system and an in vitro enzyme assay to examine wild-type (wt) and YMDD-mutant polymerases. We calculated the affinities of wt and YMDD-mutant polymerases for each natural deoxyribonucleoside triphosphate (dNTP) and determined the intracellular concentrations of each dNTP in HepG2 cells under conditions that support HBV replication. In addition, inhibition constants for lamivudine triphosphate were determined for wt and YMDD-mutant polymerases. Relative to wt HBV polymerase, each of the YMDD-mutant polymerases showed increased apparent K(m) values for the natural dNTP substrates, indicating decreased affinities for these substrates, as well as increased K(i) values for lamivudine triphosphate, indicating decreased affinity for the drug. The effect of the differences in apparent K(m) values between YMDD-mutant polymerase and wt HBV polymerase could be masked by high levels of dNTP substrates (>20 microM). However, assays using dNTP concentrations equivalent to those measured in HepG2 cells under physiological conditions showed decreased enzymatic activity of YMDD-mutant polymerases relative to wt polymerase. Therefore, the decrease in replication fitness of YMDD-mutant HBV strains results from the lower affinities (increased K(m) values) of the YMDD-mutant polymerases for the natural dNTP substrates and physiological intracellular concentrations of dNTPs that are limiting for the replication of YMDD-mutant HBV strains. PMID- 11897584 TI - In vitro and in vivo activities of gatifloxacin against Mycobacterium tuberculosis. AB - Gatifloxacin (GAT) and moxifloxacin (MXF) were evaluated in vitro to determine their activities against Mycobacterium tuberculosis. GAT was subsequently compared in a dose range study to isoniazid (INH) in a murine tuberculosis model. GAT was somewhat less active than INH. GAT and MXF were evaluated in mice infected with M. tuberculosis and were found to have similar activities. GAT was studied alone and in combination with ethambutol, ethionamide (ETA), and pyrazinamide (PZA) and compared to INH and rifampin (RIF). GAT appears to have sufficient activity alone and in combination with ETA with or without PZA to merit evaluation for treatment of tuberculosis. PMID- 11897585 TI - Artemisinin pharmacokinetics and efficacy in uncomplicated-malaria patients treated with two different dosage regimens. AB - The immediate efficacies of two oral dosage regimens of artemisinin were investigated in 77 male and female adult Vietnamese falciparum malaria patients randomly assigned to treatment with either 500 mg of artemisinin daily for 5 days (group A; n = 40) or artemisinin at a dose of 100 mg per day for 2 days, with the dose increased to 250 mg per day for 2 consecutive days and with a final dose of 500 mg on the fifth day (group B; n = 37). Parasitemia was monitored every 4 h. The average parasite clearance time was longer in group B than in group A (means +/- standard deviations, 50 +/- 23 and 34 +/- 14 h, respectively; P < 0.01). Artemisinin concentrations in saliva samples obtained on days 1 and 5 were quantified by high-performance liquid chromatography. The average oral clearance, based on saliva drug concentrations in group B patients, was twofold higher than that in group A patients on day 1 (P < 0.01), with no differences in drug half lives (P = 0.40), indicating a saturable first-pass metabolism. Female patients had higher oral clearance values on day 1. Artemisinin's pharmacokinetic parameters were similar on day 5 in both groups, although a significant increase in oral clearance from day 1 to day 5 was evident. Thus, artemisinin exhibited both dose- and time-dependent pharmacokinetics. The escalating dose studied did not result in higher artemisinin concentrations toward the end of the treatment period. PMID- 11897587 TI - Genetic determinants of tetracycline resistance in Vibrio harveyi. AB - Isolates of Vibrio harveyi, a prawn pathogen, have demonstrated multiple antibiotic resistance to commonly used antimicrobial agents, such as oxytetracycline. In this paper, we describe the cloning and characterization of two tetracycline resistance determinants from V. harveyi strain M3.4L. The first resistance determinant, cloned as a 4,590-bp fragment, was identical to tetA and flanking sequences encoded on transposon Tn10 from Shigella flexneri. The second determinant, cloned as a 3,358-bp fragment in pATJ1, contains two open reading frames, designated tet35 and txr. tet35 encodes a 369-amino-acid protein that was predicted to have nine transmembrane regions. It is a novel protein which has no homology to any other drug resistance protein but has low levels of homology (28%) to Na(+)/H(+) antiporters. Transposon mutagenesis showed that tet35 and txr were required for tetracycline resistance in a heterologous Escherichia coli host. Tetracycline accumulation studies indicate that E. coli carrying tet35 and txr can function as an energy-dependent tetracycline efflux pump but is less efficient than TetA. PMID- 11897586 TI - Antifungal activities of posaconazole, ravuconazole, and voriconazole compared to those of itraconazole and amphotericin B against 239 clinical isolates of Aspergillus spp. and other filamentous fungi: report from SENTRY Antimicrobial Surveillance Program, 2000. AB - Posaconazole, ravuconazole, and voriconazole are new triazole derivatives that possess potent, broad-spectrum antifungal activity. We evaluated the in vitro activity of these investigational triazoles compared with that of itraconazole and amphotericin B against 239 clinical isolates of filamentous fungi from the SENTRY Program, including Aspergillus spp. (198 isolates), Fusarium spp. (7 isolates), Penicillium spp. (19 isolates), Rhizopus spp. (4 isolates), Mucor spp. (2 isolates), and miscellaneous species (9 isolates). The isolates were obtained from 16 different medical centers in the United States and Canada between January and December 2000. In vitro susceptibility testing was performed using the microdilution broth method outlined in the National Committee for Clinical Laboratory Standards M38-P document. Overall, posaconazole was the most active compound, inhibiting 94% of isolates at a MIC of < or = 1 microg/ml, followed by voriconazole (91%), amphotericin B (89%), ravuconazole (88%), and itraconazole (70%). All three new triazoles demonstrated excellent activity (MIC, < or = 1 microg/ml) against Aspergillus spp. (114 Aspergillus fumigatus, 22 Aspergillus niger, 13 Aspergillus flavus, 9 Aspergillus versicolor, 8 Aspergillus terreus, and 32 Aspergillus spp.): posaconazole (98%), voriconazole (98%), ravuconazole (92%), amphotericin B (89%), and itraconazole (72%). None of the triazoles were active against Fusarium spp. (MIC at which 50% of the isolates tested were inhibited [MIC(50)], >8 microg/ml) or Mucor spp. (MIC(50), >8 microg/ml). Posaconazole and ravuconazole were more active than voriconazole against Rhizopus spp. (MIC(50), 1 to 2 microg/ml versus >8 microg/ml, respectively). Based on these results, all three new triazoles exhibited promising activity against Aspergillus spp. and other less commonly encountered isolates of filamentous fungi. The clinical value of these in vitro data remains to be seen, and in vitro in vivo correlation is needed for both new and established antifungal agents. Surveillance efforts should be expanded in order to monitor the spectrum of filamentous fungal pathogens and their in vitro susceptibility as these new antifungal agents are introduced into clinical use. PMID- 11897588 TI - In vitro low-level resistance to azoles in Candida albicans is associated with changes in membrane lipid fluidity and asymmetry. AB - The present study tracks the development of low-level azole resistance in in vitro fluconazole-adapted strains of Candida albicans, which were obtained by serially passaging a fluconazole-susceptible dose-dependent strain, YO1-16 (fluconazole MIC, 16 microg ml(-1)) in increasing concentrations of fluconazole, resulting in strains YO1-32 (fluconazole MIC, 32 microg ml(-1)) and YO1-64 (MIC, 64 microg ml(-1)). We show that acquired resistance to fluconazole in this series of isolates is not a random process but is a gradually evolved complex phenomenon that involves multiple changes, which included the overexpression of ABC transporter genes, e.g., CDR1 and CDR2, and the azole target enzyme, ERG11. The sequential rise in fluconazole MICs in these isolates was also accompanied by cross-resistance to other azoles and terbinafine. Interestingly, fluorescent polarization measurements performed by using the fluorescent probe 1,6-diphenyl 1,3,5-hexatriene revealed that there was a gradual increase in membrane fluidity of adapted strains. The increase in fluidity was reflected by observed change in membrane order, which was considerably decreased (decrease in fluorescence polarization values, P value) in the adapted strain (P value of 0.1 in YO1-64, compared to 0.19 in the YO1-16 strain). The phospholipid composition of the adapted strain was not significantly altered; however, ergosterol content was reduced in YO1-64 from that in the YO1-16 strain. The asymmetrical distribution of phosphatidylethanolamine (PE) between two monolayers of plasma membrane was also changed, with PE becoming more exposed to the outer monolayer in the YO1-64 strain. The results of the present study suggest for the first time that changes in the status of membrane lipid phase and asymmetry could contribute to azole resistance in C. albicans. PMID- 11897589 TI - bla(VIM-2) cassette-containing novel integrons in metallo-beta-lactamase producing Pseudomonas aeruginosa and Pseudomonas putida isolates disseminated in a Korean hospital. AB - We investigated the phenotypic and genetic properties of metallo-beta-lactamase producing Pseudomonas isolates collected at a tertiary-care hospital in Korea since 1995. The prevalence of imipenem resistance among Pseudomonas aeruginosa isolates reached 16% in 1997, when 9% of the resistant organisms were found to produce VIM-2 beta-lactamase, a class B enzyme previously found only in P. aeruginosa isolates from Europe. VIM-2-producing isolates of Pseudomonas putida were also detected. Resistance was transferable from both these species to P. aeruginosa PAO4089Rp by filter mating, although the resistance determinant could not be found on any detectable plasmid. Serotyping showed that many of the VIM-2 producing P. aeruginosa isolates belonged to serotypes O:11 and O:12, and pulsed field gel electrophoresis of XbaI-digested genomic DNA revealed that many had identical profiles, whereas the P. putida isolates were diverse. Sequencing showed that the bla(VIM-2) genes resided as cassettes in class 1 integrons. In contrast to previous VIM-encoding integrons, the integron sequenced from a P. aeruginosa isolate had bla(VIM) located downstream of a variant of aacA4. bla(VIM) also lay in a class 1 integron in a representative P. putida strain, but the organization of this integron was different from that sequenced from the P. aeruginosa strain. In conclusion, the metallo-beta-lactamase produced by these imipenem-resistant Pseudomonas isolates was VIM-2, and the accumulation of producers reflected clonal dissemination as well as horizontal spread. Strict measures are required in order to control a further spread of resistance. PMID- 11897590 TI - In vitro antiplasmodium effects of dermaseptin S4 derivatives. AB - The 13-residue dermaseptin S4 derivative K(4)S4(1-13)a (P) was previously shown to kill intraerythrocytic malaria parasites through the lysis of the host cells. In this study, we have sought peptides that will kill the parasite without lysing the erythrocyte. To produce such peptides, 26 compounds of variable structure and size were attached to the N terminus of P and screened for antiplasmodium and hemolytic activities in cultures of Plasmodium falciparum. Results from this screen indicated that increased hydrophobicity results in amplified antiplasmodium effect, irrespective of the linearity or bulkiness of the additive. However, increased hydrophobicity also was generally associated with increased hemolysis, with the exception of two derivatives: propionyl-P (C3-P) and isobutyryl-P (iC4-P). Both acyl-peptides were more effective than P, with 50% growth inhibition at 3.8, 4.3, and 7.7 microM, respectively. The antiparasitic effect was time dependent and totally irreversible, implying a cytotoxic effect. The peptides were also investigated in parallel for their ability to inhibit parasite growth and to induce hemolysis in infected and uninfected erythrocytes. Whereas the dose dependence of growth inhibition and hemolysis of infected cells overlapped when cells were treated with P, the acyl-peptides exerted 50% growth inhibition at concentrations that did not cause hemolysis. Noticeably, the acyl derivatives, but not P, were able to dissipate the parasite plasma membrane potential and cause depletion of intraparasite potassium under nonhemolytic conditions. These results clearly demonstrate that the acyl-peptides can affect parasite viability in a manner that is dissociated from lysis of the host cell. Overall, the data indicate the potential usefulness of this strategy for development of selective peptides as investigative tools and eventually as antimalarial agents. PMID- 11897591 TI - Phenotypic susceptibilities to tenofovir in a large panel of clinically derived human immunodeficiency virus type 1 isolates. AB - Tenofovir is a nucleotide analogue human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) inhibitor, and its oral prodrug, tenofovir disoproxil fumarate, has recently been approved for the treatment of HIV-1 infection in the United States. The objective of this study was to characterize the in vitro susceptibility profiles of a large panel of clinically derived HIV-1 isolates for tenofovir. The distribution of tenofovir susceptibilities in over 1,000 antiretroviral-naive, HIV-1-infected individuals worldwide was determined using the Virco Antivirogram assay. In addition, phenotypic susceptibilities to tenofovir and other RT inhibitors were determined in a panel of nearly 5,000 recombinant HIV-1 clinical isolates from predominantly treatment-experienced patients analyzed as a part of routine drug resistance testing. Greater than 97.5% of isolates from treatment-naive patients had tenofovir susceptibilities <3 fold above those of the wild-type controls by the Antivirogram. The clinically derived panel of 5,000 samples exhibited a broad range of antiretroviral drug susceptibilities, including 69, 43, and 16% having >10-fold-decreased susceptibilities to at least one, two, and three antiretroviral drug classes, respectively. Greater than 88% of these 5,000 clinical isolates were within the threefold susceptibility range for tenofovir, and >99% exhibited <10-fold-reduced susceptibilities to tenofovir. Decreased susceptibility to tenofovir was not directly associated with resistance to other RT inhibitors; r(2) values of log log linear regression plots of susceptibility to tenofovir versus susceptibility to other RT inhibitors were <0.4. The results suggest that the majority of treatment-naive and treatment-experienced individuals harbor HIV that remains within the normal range of tenofovir susceptibilities and may be susceptible to tenofovir disoproxil fumarate therapy. PMID- 11897592 TI - Interaction between polyamines and bacterial outer membranes as investigated with ion-selective electrodes. AB - We analyzed the interaction between polyamines and the outer membrane of Escherichia coli cells using potentiometric measurements with Ca(2+), tetraphenylphosphonium (TPP(+)), and K(+) electrodes. The Ca(2+) electrode was used to examine the ability of the polyamines to release Ca(2+) from the outer membrane. The TPP(+) electrode was used to examine the ability to permeabilize the outer membrane, since the uptake of TPP(+) was enhanced when the permeability barrier of the outer membrane was disrupted. The K(+) electrode was used to examine permeabilization in the cytoplasmic membrane by monitoring the efflux of K(+) in cytosol. Although Ca(2+) release was remarkably enhanced by increasing the number of amino groups in polyamines, no TPP(+) uptake was observed with polyamines of a simple structure, such as ethylenediamine, spermidine, and spermine. TPP(+) uptake was observed when appropriate lipophilic moieties were further attached to the polyamines with three or four amino groups, indicating that the existence of bulky moieties as well as the number of amino groups is important to induce outer membrane permeabilization. Thus, 1 naphthylacetylspermine and N,N'-bis[6-[[(2-methoxyphenyl)methyl]amino]hexyl]-1,8 octanediamine (methoctramine) were especially effective in increasing the permeability of the outer membrane of E. coli cells, being comparable to polymyxin B nonapeptide, a well-known cationic peptide showing such action. PMID- 11897594 TI - Evolution of primary protease inhibitor resistance mutations during protease inhibitor salvage therapy. AB - In order to track the evolution of primary protease inhibitor (PI) resistance mutations in human immunodeficiency virus type 1 (HIV-1) isolates, baseline and follow-up protease sequences were obtained from patients undergoing salvage PI therapy who presented initially with isolates containing a single primary PI resistance mutation. Among 78 patients meeting study selection criteria, baseline primary PI resistance mutations included L90M (42% of patients), V82A/F/T (27%), D30N (21%), G48V (6%), and I84V (4%). Despite the switching of treatment to a new PI, primary PI resistance mutations present at the baseline persisted in 66 of 78 (85%) patients. D30N persisted less frequently than L90M (50% versus 100%, respectively; P < 0.001) and V82A/F/T (50% versus 81%, respectively; P = 0.05). HIV-1 isolates from 38 (49%) patients failing PI salvage therapy developed new primary PI resistance mutations including L90M, I84V, V82A, and G48V. Common combinations of primary and secondary PI resistance mutations after salvage therapy included mutations at amino acid positions 10, 82, and 46 and/or 54 in 16 patients; 10, 90, and 71 and/or 73 in 14 patients; 10, 73, 84, 90, and 46 and/or 54 in 5 patients; 10, 48, and 82 in 5 patients; and 30, 88 and 90 in 5 patients. In summary, during salvage PI therapy, most HIV-1 isolates with a single primary PI resistance mutation maintained their original mutations, and 49% developed additional primary PI resistance mutations. The persistence of L90M, V82A/F/T, G48V, and I84V during salvage therapy suggests that these mutations play a role in clinical resistance to multiple PIs. PMID- 11897593 TI - Oxazolidinone antibiotics target the P site on Escherichia coli ribosomes. AB - The oxazolidinones are a novel class of antimicrobial agents that target protein synthesis in a wide spectrum of gram-positive and anaerobic bacteria. The oxazolidinone PNU-100766 (linezolid) inhibits the binding of fMet-tRNA to 70S ribosomes. Mutations to oxazolidinone resistance in Halobacterium halobium, Staphylococcus aureus, and Escherichia coli map at or near domain V of the 23S rRNA, suggesting that the oxazolidinones may target the peptidyl transferase region responsible for binding fMet-tRNA. This study demonstrates that the potency of oxazolidinones corresponds to increased inhibition of fMet-tRNA binding. The inhibition of fMet-tRNA binding is competitive with respect to the fMet-tRNA concentration, suggesting that the P site is affected. The fMet-tRNA reacts with puromycin to form peptide bonds in the presence of elongation factor P (EF-P), which is needed for optimum specificity and efficiency of peptide bond synthesis. Oxazolidinone inhibition of the P site was evaluated by first binding fMet-tRNA to the A site, followed by translocation to the P site with EF-G. All three of the oxazolidinones used in this study inhibited translocation of fMet tRNA. We propose that the oxazolidinones target the ribosomal P site and pleiotropically affect fMet-tRNA binding, EF-P stimulated synthesis of peptide bonds, and, most markedly, EF-G-mediated translocation of fMet-tRNA into the P site. PMID- 11897595 TI - mar Operon involved in multidrug resistance of Enterobacter aerogenes. AB - We determined the sequence of the entire marRAB operon in Enterobacter aerogenes. It is functionally and structurally analogous to the Escherichia coli operon. The overexpression of E. aerogenes MarA induces a multidrug resistance phenotype in a susceptible strain, demonstrated by a noticeable resistance to various antibiotics, a decrease in immunodetected porins, and active efflux of norfloxacin. PMID- 11897596 TI - First description of Klebsiella pneumoniae harboring CTX-M beta-lactamases (CTX-M 14 and CTX-M-3) in Taiwan. AB - Klebsiella pneumoniae isolates from Taiwan medical centers (50 strains; 1998 to 2000) with a CTX-M resistance phenotype (ceftazidime susceptible and ceftriaxone or cefotaxime nonsusceptible) were selected for initial isoelectric focusing analysis. beta-Lactamases with pIs of 7.9 (n = 22) and 8.4 (n = 28) in addition to 5.4 and/or 7.6 were detected. DNA gene sequencing identified the beta lactamases with pIs of 7.9 and 8.4 as CTX-M-14 and CTX-M-3, respectively. Molecular typing suggested inter- and intrahospital clonal dissemination of these Taiwanese CTX-M-producing Klebsiella strains. PMID- 11897597 TI - Fourteen-member macrolides suppress interleukin-8 production but do not promote apoptosis of activated neutrophils. AB - A 14-member macrolide was found to inhibit interleukin-8 (IL-8) synthesis in lipopolysaccharide-stimulated neutrophils but did not accelerate apoptosis in activated neutrophils. These data suggest that 14-member macrolides achieve clinical efficacy for chronic airway diseases partly by suppressing IL-8 production by activated neutrophils, but not by enhancing apoptosis in these cells. PMID- 11897598 TI - Synergistic activities of macrolide antibiotics against Pseudomonas aeruginosa, Burkholderia cepacia, Stenotrophomonas maltophilia, and Alcaligenes xylosoxidans isolated from patients with cystic fibrosis. AB - Azithromycin and clarithromycin were paired with other antibiotics to test synergistic activity against 300 multidrug-resistant pathogens isolated from cystic fibrosis (CF) patients. Clarithromycin-tobramycin was most active against Pseudomonas aeruginosa and inhibited 58% of strains. Azithromycin-trimethoprim sulfamethoxazole, azithromycin-ceftazidime, and azithromycin-doxycycline or azithromycin-trimethoprim-sulfamethoxazole inhibited 40, 20, and 22% of Stenotrophomonas maltophilia, Burkholderia cepacia complex, and Achromobacter (Alcaligenes) xylosoxidans strains, respectively. PMID- 11897599 TI - In vitro and in vivo activities of a novel cephalosporin, BMS-247243, against organisms other than staphylococci. AB - BMS-247243, a novel cephalosporin inhibitory for methicillin-resistant staphylococci, primarily has activity against gram-positive bacteria. The activities of BMS-247243, cefotaxime, and ceftriaxone against streptococci and Streptococcus pneumoniae were similar. BMS-247243 inhibits Enterococcus faecalis but not Enterococcus faecium. BMS-247243 also inhibits many inherently vancomycin resistant species (Leuconstoc, Lactobacillus, Pediococcus) and anaerobic gram positive bacteria. PMID- 11897600 TI - Potent in vitro antimalarial activity of metacycloprodigiosin isolated from Streptomyces spectabilis BCC 4785. AB - Bioassay-guided fractionation of the extract from the fermentation broth of Streptomyces spectabilis BCC 4785 led to the isolation of three principle antimalarial agents, metacycloprodigiosin, bafilomycin A(1), and spectinabilin. Metacycloprodigiosin exhibited potent in vitro activity against Plasmodium falciparum K1, with a 50% inhibitory concentration of 0.0050 +/- 0.0010 microg/ml, while its cytotoxicity was much weaker. PMID- 11897601 TI - Susceptibilities of Mycobacterium marinum to gatifloxacin, gemifloxacin, levofloxacin, linezolid, moxifloxacin, telithromycin, and quinupristin dalfopristin (Synercid) compared to its susceptibilities to reference macrolides and quinolones. AB - The susceptibility pattern of Mycobacterium marinum was determined. Quinupristin dalfopristin and telithromycin were less active than clarithromycin. Linezolid showed good antimicrobial activity at clinically achievable concentrations. Gatifloxacin, levofloxacin, and moxifloxacin displayed activities similar to those of ciprofloxacin. Gemifloxacin was less active. The Etest method showed variable agreement with the reference method. PMID- 11897602 TI - In vitro activities of peptide deformylase inhibitors against gram-positive pathogens. AB - The activities of six peptide deformylase (PDF) inhibitors against 107 respiratory tract pathogens were studied and compared to those of ciprofloxacin and amoxicillin-clavulanate. Against Streptococcus pneumoniae, BB-83698 and BB 83815 were the most active PDF inhibitors (MIC at which 90% of the organisms tested were inhibited [MIC(90)], 0.25 microg/ml). Five of the agents showed similar activity against Moraxella catarrhalis (MIC(90), 0.12 microg/ml). All PDF inhibitors were less active against Haemophilus influenzae; BB-3497 was the most active agent (MIC(90), 2 microg/ml). Five agents were studied against Chlamydia spp. and showed activity similar to that of ciprofloxacin (MIC, 0.5 to 4 microg/ml). This study demonstrates that PDF inhibitors have the potential to be developed for the treatment of respiratory tract infections. PMID- 11897603 TI - Activity of BMS-284756, a novel des-fluoro(6) quinolone, against Staphylococcus aureus, including contributions of mutations to quinolone resistance. AB - The in vitro activity of BMS-284756 against 602 Staphylococcus aureus isolates, including 152 that were both methicillin and ciprofloxacin resistant (MIC > or = 4 microg/ml), was determined. For ciprofloxacin-susceptible and nonsusceptible isolates, the MICs at which 50% of organisms were inhibited were 0.015 and 2 microg/ml and the MICs at which 90% of organisms were inhibited were 0.03 and 4 microg/ml, respectively. PMID- 11897604 TI - Combination of quinupristin-dalfopristin (Synercid) and rifampin is highly synergistic in experimental Staphylococcus aureus joint prosthesis infection. AB - We compared the efficacies of quinupristin-dalfopristin (Q-D; 30 mg/kg of body weight every 8 h) and vancomycin (60 mg/kg twice daily), alone or in combination with rifampin (10 mg/kg twice daily), in a rabbit model of methicillin-resistant Staphylococcus aureus knee prosthesis infection. In contrast to vancomycin, Q-D significantly reduced the mean log(10) CFU per gram of bone versus that for the controls. The combination of rifampin with either Q-D or vancomycin was significantly more effective than monotherapy. PMID- 11897605 TI - Comparison of oral artesunate and dihydroartemisinin antimalarial bioavailabilities in acute falciparum malaria. AB - Plasma antimalarial activity following oral artesunate or dihydroartemisinin (DHA) treatment was measured by a bioassay in 18 patients with uncomplicated falciparum malaria. The mean antimalarial activity in terms of the bioavailability of DHA relative to that of artesunate did not differ significantly from 1, suggesting that DHA can be formulated to be an acceptable oral alternative to artesunate. PMID- 11897606 TI - TEM derivative-producing Enterobacter aerogenes strains: dissemination of a prevalent clone. AB - TEM-24 (CAZ-6) extended-spectrum beta-lactamase (ESBL) was detected in 1988 in Clermont-Ferrand, France, in Klebsiella pneumoniae (bla(TEM-24)) and Enterobacter aerogenes (bla(TEM-24b)), and since 1994, a TEM-24-producing E. aerogenes clonal strain has been observed elsewhere in the country. To determine if the spread of this clonal strain was restricted to TEM-24-producing E. aerogenes strains, 84 E. aerogenes strains (non-TEM/SHV-producing strains, TEM-1- or -2-producing strains, and different ESBL-producing strains), isolated from 1988 to 1999 in Clermont Ferrand (n = 59) and in 11 other French hospitals in 1998 (n = 25), were studied. A clonal strain was found for TEM-24- but also for TEM-3- and TEM-1- or 2 producing isolates. This study shows that there is a clonal strain dependent on acquisition of the TEM-type enzyme (TEM-24 and other TEM types). PMID- 11897607 TI - Cloning and expression of class A beta-lactamase gene blaA(BPS) in Burkholderia pseudomallei. AB - The beta-lactamase gene blaA(BPS) in Burkholderia pseudomallei was cloned and expressed in Escherichia coli. BPS-1 is a cephalosporinase with an isoelectric point of 7.7. Sequence analysis of BPS-1 revealed conserved motifs typical of class A beta-lactamases and a relationship to the PenA (in B. cepacia) and BlaI (in Yersinia enterocolitica) lineages. PMID- 11897608 TI - Comparative in vitro activities of ertapenem (MK-0826) against 469 less frequently identified anaerobes isolated from human infections. AB - We studied the in vitro activity of ertapenem against 469 less frequently identified anaerobes from 11 genera and 52 species isolated from human infections. Ertapenem was uniformly active against 460 of 469 (98%) strains at concentrations of < or = 4 microg/ml. Only 4 of 14 Clostridium difficile, 1 of 11 Clostridium innocuum, and 4 of 6 Lactobacillus sp. strains required ertapenem concentrations of > or = 8 microg/ml for inhibition. PMID- 11897609 TI - In vitro activities of a new des-fluoroquinolone, BMS 284756, and seven other antimicrobial agents against 151 isolates of Eikenella corrodens. AB - The des-fluoroquinolone BMS 284756 was active in vitro against all 151 clinical strains of Eikenella corrodens at a MIC of < or = 0.25 microg/ml and was comparable in activity to moxifloxacin and levofloxacin. The MIC at which 90% of the isolates were inhibited by penicillin G was 2 microg/ml; MICs for 8.6% of the strains (13 of 151) were > or = 4 microg/ml, including for two beta-lactamase producing isolates. Amoxicillin-clavulanate and ampicillin-sulbactam inhibited all strains at a MIC of < or = 1 microg/ml. PMID- 11897611 TI - Novel type of staphylococcal cassette chromosome mec identified in community acquired methicillin-resistant Staphylococcus aureus strains. AB - We identified a new type of staphylococcal cassette chromosome mec (SCCmec) from two community-acquired methicillin-resistant Staphylococcus aureus (MRSA) strains. The novel element, designated type IV SCCmec, had a unique combination of the class B mec gene complex and the type 2 ccr gene complex and was much smaller in size (21 to 24 kb) than previously identified SCCmec elements of hospital-acquired MRSA. Consistent with the strains' susceptibilities to various non-beta-lactam antibiotics, the type IV SCCmec was devoid of any antibiotic resistance genes other than the mecA gene. PMID- 11897610 TI - In vitro antifungal susceptibilities of Trichosporon species. AB - The in vitro activities of amphotericin B, itraconazole, fluconazole, voriconazole, posaconazole, and ravuconazole against 39 isolates of Trichosporon spp. were determined by the NCCLS M27-A microdilution method. The azoles tested appeared to be more potent than amphotericin B. Low minimal fungicidal concentration/MIC ratios were observed for voriconazole, posaconazole, and ravuconazole, suggesting fungicidal activity. PMID- 11897612 TI - Lack of catheter infection by the efg1/efg1 cph1/cph1 double-null mutant, a Candida albicans strain that is defective in filamentous growth. AB - The molecular controls regulating the successful colonization of Candida albicans on foreign materials are not known. Here we show that a mutant C. albicans strain defective in filamentous growth and lacking the transcription factors Efg1p and Cph1p has a profoundly deficient potential for colonizing on polyurethane catheters. PMID- 11897613 TI - PCR-restriction fragment length polymorphism can also detect point mutation A2142C in the 23S rRNA gene, associated with Helicobacter pylori resistance to clarithromycin. PMID- 11897614 TI - First description of a TEM-30 (IRT-2)-producing Klebsiella oxytoca isolate. PMID- 11897615 TI - New antimicrobial agents approved by the U.S. Food and Drug Administration in 2001 and new indications for previously approved agents. PMID- 11897616 TI - Nuclear receptors. I. PPAR gamma in the gastrointestinal tract: gain or pain? AB - The peroxisome proliferator-activated receptor gamma (PPAR gamma) has recently been implicated in the pathogenesis of inflammatory bowel disease (IBD) and colon cancer. The observation that PPAR gamma agonists, through immune modulation, protect against inflammatory processes in the intestine justified their expedient evaluation in the clinical management of IBD. PPAR gamma agonists are reported to have both tumor-promoting and -inhibiting effects in models of colon cancer. These differences can, in part, be explained by PPAR gamma-independent effects of PPAR gamma agonists and by differences in the models used. Because it is still unclear how PPAR gamma impacts on colon cancer, careful monitoring of patients receiving PPAR gamma agonists and additional basic research is indicated before recommendations on the use of PPAR gamma ligands in colon cancer can be made. PMID- 11897617 TI - 85-kDa cPLA(2) plays a critical role in PPAR-mediated gene transcription in human hepatoma cells. AB - In an effort to understand the role of key eicosanoid-forming enzymes in the activation of peroxisome proliferator-activated receptor (PPAR), this study was designed to evaluate the possible contributions of cytosolic phospholipase A(2) (cPLA(2)) and group IIA secretory phospholipase A(2) (sPLA(2)) in the regulation of PPAR-mediated gene transcription in a human hepatoma cell line (HepG2). The HepG2 cells express both PPAR-alpha and -gamma but not PPAR-beta. Overexpression of cPLA(2), but not group IIA sPLA(2) in the HepG2 cells, caused a significantly increased PPAR-alpha/gamma-mediated reporter activity. Antisense inhibition of cPLA(2) resulted in a significantly decreased PPAR-alpha/gamma activity. The PPAR alpha/gamma-induced gene transcription in the HepG2 cells was inhibited by the cPLA(2) inhibitors methyl arachidonyl fluorophosphonate and arachidonyltrifluoromethyl ketone, but not by the sPLA(2) inhibitor LY311727. The expression of PPAR-alpha-mediated endogenous gene apolipoprotein A-II was increased in cells with overexpression of cPLA(2), decreased in cells with antisense inhibition of cPLA(2), but unaltered in cells with overexpression of group IIA sPLA(2). The above results demonstrated an important role of cPLA(2), but not group IIA sPLA(2) in the control of PPAR activation. The cPLA(2)-mediated PPAR activation was likely mediated by arachidonic acid and prostaglandin E(2). This study reveals a novel intracellular function of cPLA(2) in PPAR activation in HepG2 cells. The cPLA(2) thus may represent a potential therapeutic target for the control of PPAR-related liver and metabolic disorders such as obesity, lipid metabolic disorders, diabetes mellitus, and atherosclerosis. PMID- 11897618 TI - Intestinal expression of genes involved in iron absorption in humans. AB - Hereditary hemochromatosis (HHC) is one of the most frequent genetic disorders in humans. In healthy individuals, absorption of iron in the intestine is tightly regulated by cells with the highest iron demand, in particular erythroid precursors. Cloning of intestinal iron transporter proteins provided new insight into mechanisms and regulation of intestinal iron absorption. The aim of this study was to assess whether, in humans, the two transporters are regulated in an iron-dependent manner and whether this regulation is disturbed in HHC. Using quantitative PCR, we measured mRNA expression of divalent cation transporter 1 (DCT1), iron-regulated gene 1 (IREG1), and hephaestin in duodenal biopsy samples of individuals with normal iron levels, iron-deficiency anemia, or iron overload. In controls, we found inverse relationships between the DCT1 splice form containing an iron-responsive element (IRE) and blood hemoglobin, serum transferrin saturation, or ferritin. Subjects with iron-deficiency anemia showed a significant increase in expression of the spliced form, DCT1(IRE) mRNA. Similarly, in subjects homozygous for the C282Y HFE mutation, DCT1(IRE) expression levels remained high despite high serum iron saturation. Furthermore, a significantly increased IREG1 expression was observed. Hephaestin did not exhibit a similar iron-dependent regulation. Our data show that expression levels of human DCT1 mRNA, and to a lesser extent IREG1 mRNA, are regulated in an iron dependent manner, whereas mRNA of hephaestin is not affected. The lack of appropriate downregulation of apical and basolateral iron transporters in duodenum likely leads to excessive iron absorption in persons with HHC. PMID- 11897619 TI - Mechanisms of hypothermic protection against ischemic liver injury in mice. AB - Hepatic hypothermia can safely prolong the duration of hepatic inflow occlusion during complex liver resectional surgeries. The mechanism(s) by which hypothermia protects against this form of liver ischemia-reperfusion injury are not completely understood. In this study, we sought to determine whether hypothermia protects against ischemia-reperfusion injury by altering the hepatic inflammatory response. Mice undergoing 90 min of partial hepatic ischemia followed by up to 8 h of reperfusion had their body temperatures regulated at 35-37 degrees C (normothermic) or unregulated, in which rectal temperature dropped as low as 25 degrees C by the end of ischemia (hypothermic). Hypothermic mice had less liver injury vs. normothermic mice, as assessed histologically, by serum transaminase levels (89% decrease), and by liver wet-to-dry weight ratios (91% decrease). Neutrophil accumulation was absent in hypothermic mice (99% reduction vs. normothermic mice). Production of the proinflammatory cytokines tumor necrosis factor-alpha, interleukin-1 beta, and macrophage inflammatory protein-2 were reduced by up to 92%. Activation of the transcription factor nuclear factor-kappa B was not reduced in hypothermic mice, but activation of c-Jun NH(2)-terminal kinase (JNK) and the transcription factor activator protein (AP)-1 were greatly diminished. These data suggest that hypothermia suppresses the hepatic inflammatory response through selective inhibition of JNK and AP-1. PMID- 11897620 TI - Thyroid hormone regulates the activity and expression of the peptide transporter PEPT1 in Caco-2 cells. AB - An oligopeptide transporter (PEPT1) in the small intestine plays an important role in the absorption of small peptides and peptide-like drugs. We examined the effect of thyroid hormone 3,5,3'-L-triiodothyronine (T(3)) on the activity and expression of PEPT1 in human intestinal Caco-2 cells. Treatment of Caco-2 cells with T(3) inhibited [(14)C]glycylsarcosine uptake in a time- and dose-dependent manner. [(14)C]glycylsarcosine uptake was reduced by pretreatment of the cells with 100 nM T(3) for 4 days (67% of control value), whereas methyl-alpha-D-[U (14)C]glucopyranoside and [(3)H]threonine uptake were not decreased. Kinetic analysis showed that T(3) treatment significantly decreased the maximum uptake (V(max)) value for [(14)C]glycylsarcosine uptake but had no effect on the K(m) value. Moreover, T(3) treatment caused a significant decrease in the amount of PEPT1 mRNA (25% of the control). Western blotting indicated that the amount of PEPT1 protein in the apical membrane was decreased (70% of the control). These findings indicate that T(3) treatment inhibits the uptake of [(14)C]glycylsarcosine by decreasing the transcription and/or stability of PEPT1 mRNA. PMID- 11897621 TI - Somatostatin sst(2) receptors inhibit peristalsis in the rat and mouse jejunum. AB - Somatostatin [somatotropin release-inhibitory factor (SRIF)] has widespread actions throughout the gastrointestinal tract, but the receptor mechanisms involved are not fully characterized. We have examined the effect of selective SRIF-receptor ligands on intestinal peristalsis by studying migrating motor complexes (MMCs) in isolated segments of jejunum from rats, mice, and sst(2) receptor knockout mice. MMCs were recorded in 4- to 5-cm segments of jejunum mounted horizontally in vitro. MMCs occurred in rat and mouse jejunum with intervals of 104.4 +/- 10 and 131.2 +/- 8 s, respectively. SRIF, octreotide, and BIM-23027 increased the interval between MMCs, an effect fully or partially antagonized by the sst(2)-receptor antagonist Cyanamid154806. A non-sst(2) receptor-mediated component was evident in mouse as confirmed by the observation of an inhibitory action of SRIF in sst(2) knockout tissue. Blocking nitric oxide generation abolished the response to SRIF in rat but not mouse jejunum. sst(2) Receptors mediate inhibition of peristalsis in both rat and mouse jejunum, but a non-sst(2) component also exists in the mouse. Nitrergic mechanisms are differentially involved in rat and mouse jejunum. PMID- 11897622 TI - In vivo absorption of water and electrolytes in mouse intestine. Application to villin -/- mice. AB - This study was done to establish and validate a single-pass perfusion method for measuring the absorption of water and electrolytes by the mouse small intestine. The method was then used to study intestinal absorption in mice whose villin gene had been invalidated (v-/-). The single-pass perfusion of the jejunum measures the absorption of water, Cl(-), Na(+), K(+), HCO, and glucose in anesthetized wild-type and v-/- mice in vivo. We measured absorption under basal and stimulated conditions (carbachol, vasoactive intestinal polypeptide, intralumen PGE(2)). Basal absorption and stimulated secretions were similar to those previously obtained in rats. There was no difference between wild-type and v-/- mice in animals with mixed genetic background or in pure C57BL6 mice. We conclude that this in vivo perfusion method is suitable for studying the absorption/secretion of electrolytes in the mouse intestine and that a lack of villin does not significantly alter basal and secretagogue-stimulated electrolyte movements across the epithelium of the mouse jejunum in vivo. PMID- 11897623 TI - Ethanol modulates gut ischemia/reperfusion-induced liver injury in rats. AB - Whereas both ethanol and gut ischemia/reperfusion (I/R) are known to alter hepatic microvascular function, little is known about the influence of ethanol consumption on the hepatic microvascular responses to I/R. The objective of this study was to determine whether acute ethanol administration exacerbates the hepatic microvascular dysfunction induced by gut I/R. Rats were exposed to gut ischemia for 30 min followed by reperfusion. Intravital videomicroscopy was used to monitor leukocyte recruitment and the number of nonperfused sinusoids (NPS). Plasma alanine aminotransferase (ALT), tumor necrosis factor-alpha (TNF-alpha), and endotoxin concentrations were monitored. In separate experiments, ethanol was administered 15 min or 24 h before gut ischemia. In control rats, gut I/R increased the number of stationary leukocytes and NPS. It also elevated the plasma ALT, TNF-alpha, and endotoxin with a corresponding increase in intestinal mucosal permeability. Low-dose ethanol consumption 15 min before gut ischemia blunted the gut I/R-induced leukostasis and elevations in plasma TNF-alpha and ALT. However, high-dose ethanol consumption aggravated the gut I/R-induced increases in leukostasis and increases in plasma endotoxin and ALT. When ethanol was administered 24 h before, high-dose ethanol aggravated the gut I/R-induced hepatocellular injury, but low-dose ethanol did not have any effects on it. These results suggest that low-dose ethanol consumption shortly before gut ischemia attenuates the hepatic inflammatory responses, microvascular dysfunction, and hepatocellular injury elicited by gut I/R, whereas high-dose ethanol consumption appears to significantly aggravate these gut I/R-induced responses. PMID- 11897624 TI - Endothelin receptor blockers reduce I/R-induced intestinal mucosal injury: role of blood flow. AB - The aim of the present study was to assess the role of endothelin (ET) in ischemia-reperfusion (I/R)-induced mucosal injury. Mucosal permeability ((51)Cr EDTA clearance) and tissue myeloperoxidase (MPO) activity were significantly increased after 30 min of ischemia followed by 30 min of reperfusion. The I/R induced increases in mucosal permeability and polymorphonuclear leukocyte (PMN) infiltration were significantly attenuated by pretreatments with ET(A) (BQ-485) and/or ET(B) (BQ-788) receptor antagonists. Monoclonal antibody (MAb) directed against intercellular adhesion molecule-1 (ICAM-1; MAb 1A29) and superoxide dismutase (SOD) pretreatments significantly attenuated the increased mucosal permeability and PMN infiltration in a similar manner as with ET receptor antagonists. Superior mesenteric artery blood flow was significantly reduced during the reperfusion period. Both ET receptor antagonists caused a significant rise in blood flow compared with an untreated I/R group. In conclusion, our data suggest that ET(A) and/or ET(B) receptors, ICAM-1, and superoxide play an important role in I/R-induced mucosal dysfunction and PMN infiltration. Furthermore, ET is involved in the pathogenesis of post-reperfusion-induced damage and beneficial effects of ET receptor antagonism are related to an improvement of disturbed blood flow during the reperfusion period. PMID- 11897625 TI - Increased renal expression of bilirubin glucuronide transporters in a rat model of obstructive jaundice. AB - Regulation of bilirubin glucuronide transporters during hyperbilirubinemia in hepatic and extrahepatic tissues is not completely clear. In the present study, we evaluated the regulation of the bilirubin glucuronide transporters, multidrug resistance-associated proteins (MRP)2 and 3, in rats with obstructive jaundice. Bile duct ligation (BDL) or sham operation was performed in Wistar rats. Liver and kidneys were removed 1, 3, and 5 days after BDL (n = 4, in each group). Serum and urine were collected to measure bilirubin levels just before animal killing. MRP2 And MRP3 mRNA expressions were determined by real-time RT-PCR. Protein expression of MRP2 and MRP3 was determined by Western blotting. Renal MRP2 function was evaluated by para-aminohippurate (PAH) clearance. The effect of conjugated bilirubin, unconjugated bilirubin, human bile, and sulfate-conjugated bile acid on MRP2 gene expression was also evaluated in renal and hepatocyte cell lines. Serum bilirubin and urinary bilirubin excretion increased significantly after BDL. In the liver, the mRNA expression of MRP2 decreased 59, 86, and 82%, and its protein expression decreased 25, 74, and 93% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. In contrast, the liver expression of MRP3 mRNA increased 138, 2,137, and 3,295%, and its protein expression increased 560, 634, and 612% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. On the other hand, in the kidneys, the mRNA expression of MRP2 increased 162, 73, and 21%, and its protein expression increased 387, 558, and 472% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. PAH clearance was significantly increased after BDL. The mRNA expression of MRP2 increased in renal proximal tubular epithelial cells after treatment with conjugated bilirubin, sulfate-conjugated bile acid or human bile. Upregulation of MRP2 in the kidneys and MRP3 in the liver may be a compensatory mechanism to improve bilirubin clearance during obstructive jaundice. PMID- 11897626 TI - Chloride transport in rabbit esophageal epithelial cells. AB - We investigated Cl(-) transport pathways in the apical and basolateral membranes of rabbit esophageal epithelial cells (EEC) using conventional and ion-selective microelectrodes. Intact sections of esophageal epithelium were mounted serosal or luminal side up in a modified Ussing chamber, where transepithelial potential difference and transepithelial resistance could be determined. Microelectrodes were used to measure intracellular Cl(-) activity (a), basolateral or apical membrane potentials (V(mBL) or V(mC)), and the voltage divider ratio. When a basal cell was impaled, V(mBL) was -73 +/- 4.3 mV and a(i)(Cl) was 16.4 +/- 2.1 mM, which were similar in presence or absence of bicarbonate. Removal of serosal Cl(-) caused a transient depolarization of V(mBL) and a decrease in a(i)(Cl) of 6.5 +/- 0.9 mM. The depolarization and the rate of decrease of a(i)(Cl) were inhibited by approximately 60% in the presence of the Cl(-)-channel blocker flufenamate. Serosal bumetanide significantly decreased the rate of change of a(i)(Cl) on removal and readdition of serosal Cl(-). When a luminal cell was impaled, V(mC) was -65 +/- 3.6 mV and a was 16.3 +/- 2.2 mM. Removal of luminal Cl(-) depolarized V(mC) and decreased a by only 2.5 +/- 0.9 mM. Subsequent removal of Cl(-) from the serosal bath decreased a(i)(Cl) in the luminal cell by an additional 6.4 +/- 1.0 mM. A plot of V(mBL) measurements vs. log a(i)(Cl)/log a(o)(Cl) (a(o)(Cl) is the activity of Cl(-) in a luminal or serosal bath) yielded a straight line [slope (S) = 67.8 mV/decade of change in a(i)(Cl)/a(o)(Cl)]. In contrast, V(mC) correlated very poorly with log a/a (S = 18.9 mV/decade of change in a/a). These results indicate that 1) in rabbit EEC, a(i)(Cl) is higher than equilibrium across apical and basolateral membranes, and this process is independent of bicarbonate; 2) the basolateral cell membrane possesses a conductive Cl(-) pathway sensitive to flufenamate; and 3) the apical membrane has limited permeability to Cl(-), which is consistent with the limited capacity for transepithelial Cl(-) transport. Transport of Cl(-) at the basolateral membrane is likely the dominant pathway for regulation of intracellular Cl(-). PMID- 11897627 TI - Mechanism of n-butyrate uptake in the human proximal colonic basolateral membranes. AB - Current studies were undertaken to characterize the mechanism of short-chain fatty acid (SCFA) transport in isolated human proximal colonic basolateral membrane vesicles (BLMV) utilizing a rapid-filtration n-[(14)C]butyrate uptake technique. Human colonic tissues were obtained from mucosal scrapings from organ donor proximal colons. Our results, consistent with the existence of a HCO(3)( )/SCFA exchanger in these membranes, are summarized as follows: 1) n [(14)C]butyrate influx was significantly stimulated into the vesicles in the presence of an outwardly directed HCO(3)(-) and an inwardly directed pH gradient; 2) n-[(14)C]butyrate uptake was markedly inhibited (approximately 40%) by anion exchange inhibitor niflumic acid (1 mM), but SITS and DIDS (5 mM) had no effect; 3) structural analogs e.g., acetate and propionate, significantly inhibited uptake of HCO(3)(-) and pH-gradient-driven n-[(14)C]butyrate; 4) n [(14)C]butyrate uptake was saturable with a K(m) for butyrate of 17.5 +/- 4.5 mM and a V(max) of 20.9 +/- 1.2 nmol x mg protein(-1) x 5 s(-1); 5) n [(14)C]butyrate influx into the vesicles demonstrated a transstimulation phenomenon; and 6) intravesicular or extravesicular Cl(-) did not alter the anion stimulated n-[(14)C]butyrate uptake. Our results indicate the presence of a carrier-mediated HCO(3)(-)/SCFA exchanger on the human colonic basolateral membrane, which appears to be distinct from the previously described anion exchangers in the membranes of colonic epithelia. PMID- 11897628 TI - Identification of the biomechanical factors associated with the perception of distension in the human esophagus. AB - Current techniques used to investigate the mechanisms responsible for the sensory responses to distension of the human esophagus provide limited information because the degree of circumferential stretch required to determine tension can only be inferred. We used impedance planimetry to measure the cross-sectional area during esophageal distension to ascertain the degree of stretch and tension that initiated motor and sensory responses. Hyoscine-N-butyl bromide (HBB), a cholinergic muscarinic receptor blocker, was also used to alter esophageal tension during distension. Motor activity was initiated at a lower degree of stretch and tension than that which initiated sensory awareness; both increased directly with increasing distension. HBB reduced both esophageal motility and tension during distension without altering the relationship between sensation intensity and cross-sectional area. Esophageal stretch, rather than tension, thus appears to be the major factor influencing sensory responses to esophageal distension. PMID- 11897629 TI - rHuKGF ameliorates symptoms in DSS and CD4(+)CD45RB(Hi) T cell transfer mouse models of inflammatory bowel disease. AB - There is an acute need for effective therapy for inflammatory bowel disease (IBD), particularly at the level of repair of the damaged epithelium. We evaluated the efficacy of recombinant human keratinocyte growth factor (rHuKGF) in both the dextran sodium sulfate (DSS) and the CD4(+)CD45RB(Hi) T cell transfer models of IBD. Disease was induced either by the ad libitum administration to normal mice of 4% DSS in the drinking water or by the injection of 4 x 10(5) CD4(+)CD45RB(Hi) T cells into immunodeficient scid/scid mice. rHuKGF was administered by subcutaneous injection at doses of 1.0 or 3.0 mg/kg in both preventative and therapeutic regimens during both studies. rHuKGF significantly improved survival and body weight loss in the DSS model in both preventative and therapeutic dosing regimens. It also improved diarrhea, hematochezia, and hematological parameters, as well as large intestine histopathology. In the T cell transfer model, rHuKGF improved body weight loss, diarrhea, and levels of serum amyloid A, as well as large intestine histopathology. In both models of IBD, the colonic levels of intestinal trefoil factor (ITF) were elevated by the disease state and further elevated by treatment with rHuKGF. These data suggest that rHuKGF may prove useful in the clinical management of IBD and its effects are likely mediated by its ability to locally increase the levels of ITF. PMID- 11897630 TI - An evolutionarily ancient Oatp: insights into conserved functional domains of these proteins. AB - Cellular uptake of organic solutes is mediated in large part by a gene family of membrane transporters called OATPs (SLC21A). To study the structural determinants and evolutionary development of the SLC21A family, we have cloned and functionally characterized a highly expressed evolutionarily primitive Oatp from the liver of the small skate, Raja erinacea. A full-length cDNA (2.3 kb) was obtained that encodes a protein of 689 amino acids. The characteristics of this novel skate Oatp, including tissue expression, subcellular localization, substrate selectivity, Na(+) dependence, and inhibitor selectivity were generally similar to liver-specific human OATP-C and rat Oatp4. However, sequence comparisons with other OATPs indicate that this skate Oatp shares only approximately 40-50% amino acid identity with the liver-specific OATPs/Oatps and with human OATP-F. Further computer analysis revealed that the highest amino acid identities reside in the first external (78%) and internal loops (75%) and transmembrane domains 2 (76%), 3 (62%), 4 (70%), and 11 (64%). We propose that the conserved regions of the SLC21A transporter family may be critical structural determinants of substrate specificity and function. PMID- 11897631 TI - Electrical charge on protein regulates its absorption from the rat small intestine. AB - The effect of the electrical charge on the intestinal absorption of a protein was studied in normal adult rats. Chicken egg lysozyme (Lyz), a basic protein with a molecular weight of 14,300, was selected and several techniques for chemical modification were applied. Then the intestinal absorption of Lyz derivatives was evaluated by measuring the radioactivity in plasma and tissues, after the administration of an (111)In-labeled derivative to an in situ closed loop of the jejunum. After the administration of (111)In-Lyz, the level of radioactivity in plasma was comparable with the lytic activity of Lyz, supporting the fact that the radioactivity represents intact Lyz. (111)In-cationized Lyz showed a 2-3 times higher level of radioactivity in plasma, whereas the radioactivity of (111)In-anionized Lyz was much lower. The absorption rate of (111)In-Lyz derivatives calculated by a deconvolution method was correlated for the strength of their positive net charge. A similar relationship was observed using superoxide dismutase. These findings indicate that the intestinal absorption of a protein is, at least partially, determined by its electrical charge. PMID- 11897632 TI - Expression and localization of the multidrug resistance-associated protein 3 in rat small and large intestine. AB - Multidrug resistance-associated protein 3 (MRP3; symbol ABCC3), has been shown to mediate ATP-dependent transport of organic anions including 17beta-glucuronosyl estradiol, glucuronosyl bilirubin, monovalent, and sulfated bile salts. MRP3 mRNA expression was reported in rat intestine suggesting a role of MRP3 in the intestinal transport. We examined the expression and localization of MRP3 in rat small and large intestine by RT-PCR, immunofluorescence, and immunoblot analysis. MRP3 was identified in all intestinal segments by RT-PCR. MRP3 expression was low in duodenum and jejunum but markedly increased in ileum and colon. With the use of a rat MRP3 specific antibody, MRP3 was localized to the basolateral domains of enterocytes. Immunofluorescence analysis and immunoblot analysis confirmed a strong expression of rat MRP3 in ileum and colon. In contrast, MRP2 was predominantly expressed in the proximal segments of rat small intestine. Our findings demonstrate a high expression of rat MRP3 in ileum and colon and provide evidence for an involvement of MRP3 in the ATP-dependent transport of organic anions, including bile salts from the enterocyte into blood. PMID- 11897633 TI - Putative effect of Helicobacter pylori and gastritis on gastric acid secretion in cat. AB - Helicobacter pylori may increase or inhibit gastric acid. We studied acid variations and plasma gastrin in cats harboring Helicobacter felis, harboring H. pylori, or free of gastric pathogens with reference to thioperamide (H(3) receptor antagonist) and SR-27417A (PAF receptor antagonist). In cats harboring H. felis, gastric mucosa were histologically normal. After H. felis eradication, pentagastrin-stimulated acid secretion was increased (40%) compared with the situation before eradication. Thioperamide abolished this inhibitory effect of H. felis, whereas SR-27417A did not. Basal and meal-stimulated plasma gastrin levels were not affected by eradication therapy. Acid secretion was inhibited (-80%) in week 3, increased from weeks 5 to 9, and remained constant for up to 42 weeks after H. pylori infection. SR-27417A had no effect on acid secretion before week 8 but inhibited it thereafter, and thioperamide increased it (20%) only before week 7 in those cats. Helicobacter inhibits gastric acid via an H(3) receptor pathway. Inflammatory mediators are thus involved in adaptation to the inhibitory effects of H. pylori on acid secretion. PMID- 11897634 TI - Sound the tocsin! Beware adverse effects of lung inflammation early in gestation. PMID- 11897635 TI - Transmission of multidrug-resistant tuberculosis: limited by man or nature? PMID- 11897636 TI - When does 'mild' obstructive sleep apnea/hypopnea syndrome merit continuous positive airway pressure treatment? PMID- 11897637 TI - Across-country viewpoints on sleepiness during driving. PMID- 11897638 TI - Can health care costs be reduced by limiting intensive care at the end of life? PMID- 11897639 TI - Cowbird research and measuring pulsatile diffusing capacity. PMID- 11897640 TI - Mutations of the cystic fibrosis gene and intermediate sweat chloride levels in children. AB - The incidence of mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in children with intermediate sweat chloride levels is unknown. The results of 2,349 sweat tests performed at two Belgian university hospitals were reviewed. Intermediate chloride concentrations were observed in 98 subjects (4.2%), 68 being younger than 18 years of age. Forty-three children could be traced and their parents agreed to take part in the study. Exhaustive analysis of the CFTR gene disclosed a total of 24 putative mutations (27.9%). Three subjects were found to carry only one CFTR mutation, whereas 10 harbored one mutation on both CFTR genes. These 10 children were investigated in detail. At the time of writing, the mean age (+/-SD) of this group is 8.9 years (+/-4.2 years). Nine children are pancreatic sufficient. Three have been asymptomatic for more than two years, whereas the others display, to different degrees, clinical features suggestive of CF. The sweat chloride concentration is slightly higher in this group (39.4 +/- 5.4 mM) than in subjects without CFTR mutation (35.2 +/- 4.4 mM, p < 0.05). The nasal potential difference was abnormal in five of the nine subjects tested. In this study, 23% of children displaying intermediate sweat chloride levels were found to carry a putative mutation on both CFTR genes. PMID- 11897641 TI - The relationship between genotype and exercise tolerance in children with cystic fibrosis. AB - The relationship between fitness and genotype in children with cystic fibrosis (CF) and at least one copy of the DeltaF508 mutation was examined. Genotype was classified according to the second CF mutation. Fitness was measured by peak aerobic capacity (using a modified Bruce protocol during treadmill exercise) and anaerobic power (using the Wingate test on a cycle ergometer). The class of cystic fibrosis transmembrane regulator proteins (CFTR) mutation was statistically related with aerobic capacity, peak anaerobic power, body mass index, lung function (forced expiratory volume in one second), and disease severity as measured by the Shwachman score. Patients with mutations causing defective CFTR production (Class I) or processing (Class II) had a significantly lower peak aerobic capacity (28.6 +/- 4.2 ml/kg/min and 31.7 +/- 5.4 ml/kg/min, respectively) than those with a mutation conferring defective regulation of CFTR (Class III) (43.9 +/- 6.4 ml/kg/min). The peak anaerobic power in subjects with mutations inducing decreased CFTR conduction (Class IV) or CFTR mRNA (Class V), were significantly higher (11.4 +/- 1.7 and 11.6 +/- 1.5 watts/kg, respectively) than children with Class I (9.7 +/- 1.4 watts/kg), Class II (9.8 +/- 1.4 watts/kg), or Class III (10.5 +/- 1.8 watts/kg) mutations. There were no statistically significant differences in the lung function of patients with the different mutations. These results indicate a relationship between CF genotype and some measures of fitness, the mechanisms of which remain to be determined. PMID- 11897642 TI - Hospitalized community-acquired pneumonia in the elderly: age- and sex-related patterns of care and outcome in the United States. AB - Community-acquired pneumonia (CAP) is a frequent cause of hospital admission and death among elderly patients, but there is little information on age- and sex specific incidence, patterns of care (intensive care unit admission and mechanical ventilation), resource use (length of stay and hospital costs), and outcome (mortality). We conducted an observational cohort study of all Medicare recipients, aged 65 years or older, hospitalized in nonfederal U.S. hospitals in 1997, who met ICD-9-CM-based criteria for CAP. We identified 623,718 hospital admissions for CAP (18.3 per 1,000 population > or = 65 years), of which 26,476 (4.3%) were from nursing homes and of which 66,045 (10.6%) died. The incidence rose five-fold and mortality doubled as age increased from 65-69 to older than 90 years. Men had a higher mortality, both unadjusted (odds ratio [OR]: 1.21 [95% CI: 1.19-1.23]) and adjusted for age, location before admission, underlying comorbidity, and microbiologic etiology (OR: 1.15 [95% CI: 1.13-1.17]). Mean hospital length of stay and costs per hospital admission were 7.6 days and $6,949. For those admitted to the intensive care unit (22.4%) and for those receiving mechanical ventilation (7.2%), mean length of stay and costs were 11.3 days and $14,294, and 15.7 days and $23,961, respectively. Overall hospital costs were $4.4 billion (6.3% of the expenditure in the elderly for acute hospital care), of which $2.1 billion was incurred by cases managed in intensive care units. We conclude that in the hospitalized elderly, CAP is a common and frequently fatal disease that often requires intensive care unit admission and mechanical ventilation and consumes considerable health care resources. The sex differences are of concern and require further investigation. PMID- 11897643 TI - A randomized controlled trial of continuous positive airway pressure in mild obstructive sleep apnea. AB - A common clinical dilemma faced by sleep physicians is in deciding the level of severity at which patients with obstructive sleep apnea (OSA) should be treated. There is particular uncertainty about the need for, and the effectiveness of, treatment in mild cases. To help define the role of nasal continuous positive airway pressure (CPAP) treatment in mild OSA we undertook a randomized controlled cross-over trial of CPAP in patients with an apnea- hypopnea index (AHI) of 5 - 30 (mean, 12.9 +/- 6.3 SD). Twenty-four-hour blood pressure and neurobehavioral function were measured at baseline, after 8 wk of treatment with CPAP, and after 8 wk of treatment with an oral placebo tablet. Twenty-eight of 42 patients enrolled in the study completed both treatment arms. Baseline characteristics were not different between those who completed the study and those who did not complete the study. Patients used CPAP for a mean (SD) of 3.53 (2.13) h per night and the mean AHI on the night of CPAP implementation was 4.24 (2.9). Nasal CPAP improved self-reported symptoms of OSA, including snoring, restless sleep, daytime sleepiness, and irritability (in-house questionnaire), more than did placebo, but did not improve objective (Multiple Sleep Latency Test) or subjective (Epworth Sleepiness Scale) measures of daytime sleepiness. We found no benefit of CPAP over placebo in any tests of neurobehavioral function, generic SF 36 (36-item Short Form Medical Outcomes Survey) or sleep-specific (Functional Outcomes of Sleep Questionnaire) quality of life questionnaires, mood score (Profile of Moods States and Beck Depression Index), or 24-h blood pressure. However, the placebo tablet resulted in a significant improvement in a wide range of functional variables compared with baseline. This placebo effect may account for some of the treatment responses to CPAP observed previously in patients with mild OSA. PMID- 11897644 TI - Prospective, randomized, controlled pilot study of partial liquid ventilation in adult acute respiratory distress syndrome. AB - We evaluated the safety and efficacy of partial liquid ventilation (PLV) with perflubron in adult patients with acute lung injury and the acute respiratory distress syndrome (ARDS) in a multicenter, prospective, controlled, randomized exploratory clinical trial. Ninety adult patients with PaO2/FIO2 ratios > 60 and < 300 with ARDS for no more than 24 hours were randomized to receive PLV (n = 65) with administration of perflubron through an endotracheal tube sideport or conventional mechanical ventilation (CMV, n = 25) for a maximum of five days. Although a significant reduction in progression to ARDS was noted among patients with PLV, no significant differences in the number of days free from the ventilator at 28 days (CMV = 6.7 +/- 1.8, PLV = 6.3 +/- 1.0 days, p = 0.85), the incidence of mortality (CMV = 36%, PLV = 42%, p = 0.63), or any pulmonary-related parameter were observed. During a post hoc subgroup analysis, significantly more rapid discontinuation of mechanical ventilation (p = 0.045) and a trend toward an increase in the number of days free from the ventilator at 28 days (CMV = 3.2 +/- 1.9, PLV = 8.0 +/- 2.2 days, p = 0.06) were observed during PLV among those patients under 55 years of age with acute lung injury or ARDS. Episodes of hypoxia, respiratory acidosis, and bradycardia occurred more frequently in the PLV group, but these were transient and self-limited. Further evaluation of PLV is warranted to further define beneficial effects in well-defined groups of patients and also to gain additional information regarding safety. PMID- 11897645 TI - Human leukocyte antigen Class II amino acid epitopes: susceptibility and progression markers for beryllium hypersensitivity. AB - Chronic beryllium disease (CBD) is a hypersensitivity granulomatosis characterized by beryllium hypersensitivity (BH) and mediated by CD4+ T cells. However, all individuals with BH may not develop CBD. To examine the role of the three different human leukocyte antigen (HLA) Class II isotypes in BH with (CBD) and without clinical disease (BHWCD), we performed DNA-based typing of HLA-DPB1, HLA-DQB1, and HLA-DRB1 loci on 55 subjects with BH (25 with established CBD and 30 with BHWCD), and compared this with the results for 82 beryllium-exposed workers with no evidence of BH. The allele distribution was utilized to identify candidate amino acid epitopes that differed between the study groups. HLA-DPB1 E69 was the most important marker for BH, and did not differentiate BHWCD from CBD. A significant association with CBD was observed with HLA-DQB1-G86 (p(corr) < 0.04), and HLA-DRB1-S11 was significantly increased in CBD as compared with BHWCD (p < 0.03). These observations suggest that HLA-DPB1-E69 is a marker for susceptibility to BH and not just a progression marker for CBD. In addition, HLA amino acid epitopes on HLA-DRB1 and -DQB1, in concert with or independently of HLA-DPB1-E69, may be associated with progression to CBD. PMID- 11897646 TI - Genetic loci influencing lung function: a genome-wide scan in the Framingham Study. AB - Prior studies have found cross-sectional lung function to be highly heritable. In the present study, we used a 10-cM genome-wide scan of 1,578 members of 330 families participating in the Framingham Study to test for linkage of genetic markers to level of lung function as determined by spirometry during middle age. At this age, lung function measures may reflect the effects of genes influencing lung growth and development, as well as of those influencing decline in lung function during adulthood. We performed spirometry on 345 members of the Original Cohort and 1,233 members of the Offspring Cohort of the Framingham Study. The effects of age, height, body mass index, and smoking status on spirometric measures were adjusted through linear regression models created separately for men and women in each cohort. Standardized residuals for FEV1, FVC, and the ratio of FEV1 to FVC were obtained from these models. The residual spirometric measures were analyzed for linkage to the genome scan markers through the use of variance component models in the Sequential Oligogenic Linkage Analysis Routines software program. The loci most strongly influencing FEV1 and FVC colocalized on chromosomes 4, 6, and 21. FEV1 was most influenced by the locus on chromosome 6 (logarithm of the odds favoring genetic linkage [LOD] = 2.4), whereas chromosome 21 contained markers with the strongest linkage to FVC (LOD = 2.6). PMID- 11897647 TI - Continuous subcutaneous infusion of treprostinil, a prostacyclin analogue, in patients with pulmonary arterial hypertension: a double-blind, randomized, placebo-controlled trial. AB - Pulmonary arterial hypertension is a life-threatening disease for which continuous intravenous prostacyclin has proven to be effective. However, this treatment requires a permanent central venous catheter with the associated risk of serious complications such as sepsis, thromboembolism, or syncope. Treprostinil, a stable prostacyclin analogue, can be administered by a continuous subcutaneous infusion, avoiding these risks. We conducted a 12-week, double blind, placebo-controlled multicenter trial in 470 patients with pulmonary arterial hypertension, either primary or associated with connective tissue disease or congenital systemic-to-pulmonary shunts. Exercise capacity improved with treprostinil and was unchanged with placebo; the between treatment group difference in median six-minute walking distance was 16 m (p = 0.006). Improvement in exercise capacity was greater in the sicker patients and was dose related, but independent of disease etiology. Concomitantly, treprostinil significantly improved indices of dyspnea, signs and symptoms of pulmonary hypertension, and hemodynamics. The most common side effect attributed to treprostinil was infusion site pain (85%) leading to premature discontinuation from the study in 8% of patients. Three patients in the treprostinil treatment group presented with an episode of gastrointestinal hemorrhage. We conclude that chronic subcutaneous infusion of treprostinil is an effective treatment with an acceptable safety profile in patients with pulmonary arterial hypertension. PMID- 11897648 TI - Early gestational intra-amniotic endotoxin: lung function, surfactant, and morphometry. AB - We determined the effects in preterm lambs of endotoxin-induced inflammation at early gestational ages on lung function and structure and on the surfactant system. Pregnant ewes were randomized to one of five intra-amniotic endotoxin (Escherichia coli 055:B5) groups: 1 mg injected at 60 days of gestation, 1 mg at 80 days, 1 mg at 100 days, 1 mg at 60 days plus 100 days, or 0.6 mg/ day infused from Day 80 to Day 108. Control lambs received saline treatments. At 125 days, lung function was improved in all endotoxin groups. Marked increases in saturated phosphatidylcholine in lung tissue but not alveolar lavage samples were seen in all endotoxin groups except the 60- plus 100-day group. Surfactant protein mRNA and protein pool sizes were affected differently according to the timing of endotoxin treatment, but a large increase in the amount of mature surfactant protein B in alveolar lavage samples was observed in all endotoxin groups. Lung to-body weight ratio, alveolar number, total surface area, and alveolar wall thickness were reduced by 80- to 108-day endotoxin. Intra-amniotic inflammatory stimuli in early gestation can alter pulmonary development, with the net effect of improving preterm lung function, despite changes in surfactant and lung growth that are similar to changes in the lungs of ventilated animals developing bronchopulmonary dysplasia. PMID- 11897649 TI - Limited transmission of multidrug-resistant tuberculosis despite a high proportion of infectious cases in Los Angeles County, California. AB - Preventing transmission of multidrug-resistant tuberculosis is critical because of treatment toxicity, cost, and the lack of effective therapy for latent infection. We attempted to determine the extent of transmission in Los Angeles County by comparing relatedness of multidrug-resistant tuberculosis cases using restriction fragment length polymorphism and by cross-matching contact information to the Tuberculosis Registry. Strain typing was done on isolates of 102 pulmonary multidrug-resistant cases identified between August 1993 and 1998. Seventy-one (70%) of the cases had cavitary lesions on chest radiograph, and 94 (92%) had sputa smear-positive for acid fast bacilli. Fifteen (15%) of the cases were known to be infected with human immunodeficiency virus. Four molecular clusters of two cases each and one closely related pair were identified among the 102 cases; contact investigation successfully identified all clusters but one. Among 946 contacts identified and cross-matched with the county's Tuberculosis Registry, one secondary case due to drug-resistant Mycobacterium bovis was found. To summarize, a very high proportion of pulmonary multidrug-resistant tuberculosis cases in Los Angeles County were infectious. Molecular strain typing indicated limited spread of disease, although it underestimated transmission compared with contact investigation. We believe aggressive surveillance and case management were critical to limiting the spread of multidrug- resistant tuberculosis. PMID- 11897650 TI - Luteolin reduces lipopolysaccharide-induced lethal toxicity and expression of proinflammatory molecules in mice. AB - Luteolin is a flavonoid that has been shown to reduce proinflammatory molecule expression in vitro. In the present study, we have tested the ability of luteolin to inhibit lipopolysaccharide (LPS)- induced lethal toxicity and proinflammatory molecule expression in vivo. Mice receiving LPS (Salmonella enteriditis LPS, 32 mg/kg, intraperitoneally) exhibited high mortality with only 4.1% of the animals surviving seven days after the LPS challenge. On the contrary, mice that had received luteolin (0.2 mg/kg, intraperitoneally) before LPS showed an increased survival rate with 48% remaining alive on Day 7. To investigate the mechanism by which luteolin affords protection against LPS toxicity we measured intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-alpha (TNF-alpha) production in response to LPS in the presence or absence of luteolin pretreatment. Treatment of animals with LPS increased serum TNF-alpha levels in a time-dependent manner. The increase in peak serum TNF-alpha levels was sensitive to luteolin pretreatment. Luteolin pretreatment also reduced LPS-stimulated ICAM 1 expression in the liver and abolished leukocyte infiltration in the liver and lung. We conclude that luteolin protects against LPS-induced lethal toxicity, possibly by inhibiting proinflammatory molecule (TNF-alpha, ICAM-1) expression in vivo and reducing leukocyte infiltration in tissues. PMID- 11897651 TI - The failure of interleukin-10-deficient mice to develop airway hyperresponsiveness is overcome by respiratory syncytial virus infection in allergen-sensitized/challenged mice. AB - Interleukin-10-deficient mice develop a robust pulmonary inflammatory response but no airway hyperresponsiveness (AHR) to inhaled methacholine (MCh) following allergen sensitization and challenge. In the present study, we investigated the effect of respiratory syncytial virus (RSV) infection on AHR and pulmonary inflammation in allergic IL-10-/- mice. Unlike littermate control mice, RSV infected or ovalbumin (OVA)-sensitized/challenged IL-10-/- mice failed to develop significant AHR. In contrast, sensitized/challenged IL-10-/- mice infected with RSV did develop AHR accompanied by increased eosinophil numbers, both in bronchoalveolar lavage (BAL) and pulmonary tissue, and mucin production in airway epithelium. The cytokine profile in OVA-sensitized/challenged IL-10-/- mice was skewed toward a Th1 response but after RSV infection, this response was more of a Th2 type, with increased IL-5 levels in the BAL. Studies with an RSV mutant that lacks the G and SH genes showed equal enhancement of the AHR response as the parental wild-type strain, indicating that G protein is not essential to this response. These data suggest that RSV infection can overcome the failure of development of AHR in allergic IL-10-/- mice. PMID- 11897652 TI - Endobronchial ultrasonography in the assessment of centrally located early-stage lung cancer before photodynamic therapy. AB - To evaluate the utility of endobronchial ultrasonography (EBUS) in selecting appropriate candidates with centrally located early-stage lung cancer for photodynamic therapy (PDT) with curative intent, we performed EBUS before PDT in 18 biopsy-proven squamous cell carcinomas (including three carcinoma in situ) that had been considered to be appropriate candidates for PDT by conventional bronchoscopy and high-resolution computed tomography (HR-CT). Nine lesions were diagnosed as intracartilaginous by EBUS and subsequently PDT was performed. Long term complete remission has been achieved in these patients with a median follow up term after PDT of 32 months. The remaining nine lesions were diagnosed as extracartilaginous by EBUS and were considered candidates for other therapies such as surgical resection, chemotherapy, and radiotherapy, although two were invisible by HR-CT, three were superficial, and five were < or = 1 cm in diameter on observation by bronchoscopy. The depth of tumor invasion estimated by EBUS was proven to be accurate by histopathologic findings in six specimens after surgical resection. We conclude that EBUS is a useful technique that might be considered in addition to conventional bronchoscopy and HR-CT to improve the efficacy of PDT in patients with centrally located early-stage lung cancer. PMID- 11897653 TI - Future research directions in chronic obstructive pulmonary disease. PMID- 11897654 TI - Effects of theophylline on airway eosinophils. PMID- 11897657 TI - Bleomycin-induced lung fibrosis: the authors should have used another method to induce pulmonary lesions resembling human idiopathic pulmonary fibrosis. PMID- 11897658 TI - Reprogrammed gene expression in a somatic cell-free extract. AB - We have developed a somatic cell-free system that remodels chromatin and activates gene expression in heterologous differentiated nuclei. Extracts of stimulated human T cells elicit chromatin binding of transcriptional activators of the interleukin-2 (IL-2) gene, anchoring and activity of a chromatin remodeling complex and hyperacetylation of the IL-2 promoter in purified exogenous resting T-cell nuclei. The normally repressed IL-2 gene is transcribed in nuclei from quiescent human T cells and from various non-T-cell lines. This demonstrates that somatic cell extracts can be used to reprogram gene expression in differentiated nuclei. In vitro reprogramming may be useful for investigating regulation of gene expression and for producing replacement cells for the treatment of a wide variety of diseases. PMID- 11897659 TI - Release factor 2 frameshifting sites in different bacteria. AB - The mRNA encoding Escherichia coli polypeptide chain release factor 2 (RF2) has two partially overlapping reading frames. Synthesis of RF2 involves ribosomes shifting to the +1 reading frame at the end of the first open reading frame (ORF). Frameshifting serves an autoregulatory function. The RF2 gene sequences from the 86 additional bacterial species now available have been analyzed. Thirty percent of them have a single ORF and their expression does not require frameshifting. In the approximately 70% that utilize frameshifting, the sequence cassette responsible for frameshifting is highly conserved. In the E. coli RF2 gene, an internal Shine-Dalgarno (SD) sequence just before the shift site was shown earlier to be important for frameshifting. Mutagenic data presented here show that the spacer region between the SD sequence and the shift site influences frameshifting, and possible mechanisms are discussed. Internal translation initiation occurs at the shift site, but any functional role is obscure. PMID- 11897660 TI - Phosphorylation of histone H3 is functionally linked to retinoic acid receptor beta promoter activation. AB - Ligand-dependent transcriptional activation of retinoic acid receptors (RARs) is a multistep process culminating in the formation of a multimeric co-activator complex on regulated promoters. Several co-activator complexes harbor an acetyl transferase activity, which is required for retinoid-induced transcription of reporter genes. Using murine P19 embryonal carcinoma cells, we examined the relationship between histone post-translational modifications and activation of the endogenous RARbeta2 promoter, which is under the control of a canonical retinoic acid response element and rapidly induced upon retinoid treatment. While histones H3 and H4 were constitutively acetylated at this promoter, retinoid agonists induced a rapid phosphorylation at Ser10 of histone H3. A retinoid antagonist, whose activity was independent of co-repressor binding to RAR, could oppose this agonist-induced H3 phosphorylation. Since such post-translational modifications were not observed at several other promoters, we conclude that histone H3 phosphorylation may be a molecular signature of the activated, retinoid-controlled mRARbeta2 gene promoter. PMID- 11897661 TI - Promoter protection by a transcription factor acting as a local topological homeostat. AB - Binding of the Escherichia coli global transcription factor FIS to the upstream activating sequence (UAS) of stable RNA promoters activates transcription on the outgrowth of cells from stationary phase. Paradoxically, while these promoters require negative supercoiling of DNA for optimal activity, FIS counteracts the increase of negative superhelical density by DNA gyrase. We demonstrate that binding of FIS at the UAS protects the rrnA P1 promoter from inactivation at suboptimal superhelical densities. This effect is correlated with FIS-dependent constraint of writhe and facilitated untwisting of promoter DNA. We infer that FIS maintains stable RNA transcription by stabilizing local writhe in the UAS. These results suggest a novel mechanism of transcriptional regulation by a transcription factor acting as a local topological homeostat. PMID- 11897662 TI - Human Asf1 and CAF-1 interact and synergize in a repair-coupled nucleosome assembly pathway. AB - The efficient assembly of newly replicated and repaired DNA into chromatin is essential for proper genome function. Based on genetic studies in Saccharomyces cerevisiae, the histone chaperone anti-silencing function 1 (Asf1) has been implicated in the DNA repair response. Here, the human homologs are shown to function synergistically with human CAF-1 to assemble nucleosomes during nucleotide excision repair in vitro. Furthermore, we demonstrate that hAsf1 proteins can interact directly with the p60 subunit of hCAF-1. In contrast to hCAF-1 p60, the nuclear hAsf1 proteins are not significantly associated with chromatin in cells before or after the induction of DNA damage, nor specifically recruited to damaged DNA during repair in a bead-linked DNA assay. A model is proposed in which the synergism between hAsf1 and CAF-1 for nucleosome formation during DNA repair is achieved through a transient physical interaction allowing histone delivery from Asf1 to CAF-1. PMID- 11897663 TI - Plk1 promotes nuclear translocation of human Cdc25C during prophase. AB - The nuclear accumulation of active M-phase promoting factor (MPF) during prophase is thought to be essential for coordinating M-phase events in vertebrate cells. The protein phosphatase Cdc25C, an activator of MPF, enters the nucleus to keep MPF active in the nucleus during prophase. However, the molecular mechanisms that control nuclear translocation of Cdc25C during prophase are unknown. We show that phosphorylation of a serine residue (Ser198) in a nuclear export signal sequence of human Cdc25C occurs during prophase and promotes nuclear localization of Cdc25C. We also show that Polo-like kinase 1 (Plk1) is responsible for this phosphorylation and that constitutively active Plk1 promotes nuclear localization of Cdc25C. Remarkably, a mutant Cdc25C in which Ser198 is replaced by alanine remains in the cytoplasm when wild-type Cdc25C accumulates in the nucleus during prophase. These results suggest that Plk1 phosphorylates Cdc25C on Ser198 and regulates nuclear translocation of Cdc25C during prophase. PMID- 11897664 TI - An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo. AB - pEPI-1, a vector in which a chromosomal scaffold/matrix-attached region (S/MAR) is linked to the simian virus 40 origin of replication, is propagated episomally in CHO cells in the absence of the virally encoded large T-antigen and is stably maintained in the absence of selection pressure. It has been suggested that mitotic stability is provided by a specific interaction of this vector with components of the nuclear matrix. We studied the interactions of pEPI-1 by crosslinking with cis-diamminedichloroplatinum II, after which it is found to copurify with the nuclear matrix. In a south-western analysis, the vector shows exclusive binding to hnRNP-U/SAF-A, a multifunctional scaffold/matrix specific factor. Immunoprecipitation of the crosslinked DNA-protein complex demonstrates that pEPI-1 is bound to this protein in vivo. These data provide the first experimental evidence for the binding of an artificial episome to a nuclear matrix protein in vivo and the basis for understanding the mitotic stability of this novel vector class. PMID- 11897666 TI - Are glucocorticoids good or bad for brain development and plasticity? PMID- 11897665 TI - Prothymosin alpha interacts with the CREB-binding protein and potentiates transcription. AB - Prothymosin alpha (ProTalpha) is a histone H1-binding protein localized in sites of active transcription in the nucleus. We report here that ProTalpha physically interacts with the CREB-binding protein (CBP), which is a versatile transcription co-activator. Confocal laser scanning microscopy reveals that ProTalpha partially colocalizes with CBP in discrete subnuclear domains. Using transient transfections, we show that ProTalpha synergizes with CBP and stimulates AP1- and NF-kappaB-dependent transcription. Furthermore, overexpression of ProTalpha enhances the transactivation potential of CBP. These findings reveal a new function for ProTalpha in transcription activation, probably through CBP-mediated recruitment to different promoters. PMID- 11897667 TI - Prolonged low-dose dexamethasone, in early gestation, has no long-term deleterious effect on normal ovine fetuses. AB - Low-dose dexamethasone (D) treatment is used in pregnancies where the fetus is suspected to be at risk of congenital/virilizing adrenal hyperplasia. To study if this treatment had any immediate or long-term effects in normal fetuses, pregnant ewes were treated with D (20 microg/kg maternal body weight x d) or saline (S), from d 25-45 of gestation. Tissue was collected from fetuses killed at 45 d (S = 6; D = 8), 130 d (S = 8; D = 8), or lambs at 2 months of age (S = 6; D = 6) and mRNA levels measured using real-time PCR. D treatment reduced adrenal wt at 45 d (S, 12.2 +/- 0.7 mg; D, 6.3 +/- 0.4 mg) and significantly decreased adrenal mRNA for P(450scc). At 130 d, fetuses from the D treatment were growth retarded (S, 3.2 +/- 0.1 kg; D, 2.5 +/- 0.1 g), but the adrenals were appropriate for the body weight. mRNA levels of angiotensinogen, the AT(1) receptor and mineralocorticoid receptor (MR) and GR were similar in kidney and brain (hypothalamus, hippocampus, medulla oblongata) except for hippocampal expression of MR and GR, which was significantly decreased by D treatment. By 2 months, BW and hippocampal MR and GR mRNA levels were similar, and lambs were normotensive (S, 83 +/- 3 mm Hg; D, 78 +/- 3 mm Hg). Thus, there were no persistent, long-term effects of prolonged low dose D treatment in normal ovine fetuses. PMID- 11897668 TI - Plasma leptin concentration in fetal sheep during late gestation: ontogeny and effect of glucocorticoids. AB - The ontogeny and developmental control of plasma leptin concentration in the fetus are poorly understood. The present study investigated plasma leptin concentration in chronically catheterized sheep fetuses near term, and in neonatal and adult sheep. The effect of glucocorticoids on plasma leptin in utero was examined by fetal adrenalectomy and exogenous cortisol or dexamethasone infusion. In intact, untreated fetuses studied between 130 and 140 d (term, 145 +/- 2 d), plasma leptin concentration increased in association with the prepartum cortisol surge. Positive relationships were observed between plasma leptin in utero and both gestational age and plasma cortisol. Plasma leptin was also inversely correlated with fetal p(a)O(2). The ontogenic rise in plasma leptin was abolished by fetal adrenalectomy. In intact fetuses at 123-127 d, plasma leptin was increased by infusions of cortisol (3-5 mg kg(-1)d(-1), +127 +/- 21%) for 5 d and dexamethasone (45-60 microg kg(-1)d(-1), +268 +/- 61%) for 2 d. However, the cortisol-induced rise in plasma leptin was transient; by the fifth day of infusion, plasma leptin was restored to within the baseline range. These findings show that, in the sheep fetus, an intact adrenal gland is required for the normal ontogenic rise in plasma leptin near term. Furthermore, fetal treatment with exogenous and endogenous glucocorticoids increases circulating leptin concentration in utero. PMID- 11897669 TI - TGF beta-induced Smad signaling remains intact in primary human ovarian cancer cells. AB - Disruptions in TGF beta signaling have been implicated in various human cancers, including ovarian cancer. Our goal was to determine whether ovarian cancer cells isolated from patient ascites fluid were growth inhibited by TGF beta 1 treatment and further characterize the expression and activity profile of TGF beta/Smad signaling components in human ovarian cancer cells. We found that 9 of 10 primary cultures of ovarian cancer cells (OC2-10) were growth inhibited by 16 pM TGF beta 1. One primary ovarian cancer sample (OC1) and the established ovarian cancer cell lines CaOV3 and SkOV3 continued to grow in the presence of TGF beta 1. All cells expressed components of the TGF beta/Smad signaling pathway including TGF beta 1, T beta RI, T beta RII, Smad2, -3, -4, and Smad anchor for receptor activation. Although OC1, CaOV3, and SkOV3 are not growth inhibited by TGF beta 1, they can transmit the TGF beta 1 signal to turn on a transfected TGF beta/Smad reporter gene, p3TP.lux. In addition, all cells up-regulate the endogenous TGF beta target genes Smad7 and PAI-1. p15(Ink4B) mRNA is also up-regulated with TGF beta 1 treatment in OC2-9, whereas the p15(Ink4B) gene has been deleted in OC1, CaOV3, and SkOV3 cells. Homozygous deletion of p15(Ink4B) may account for TGF beta resistance in some populations of ovarian cancer cells. Our data demonstrate that the TGF beta/Smad signaling pathway remains functional in human ovarian cancer cells and suggest that if abnormalities exist in the cellular response of TGF beta signals, they must lie downstream of the Smad proteins. PMID- 11897670 TI - Immune deviation away from Th1 in interferon-gamma knockout mice does not enhance TSH receptor antibody production after naked DNA vaccination. AB - TSH receptor (TSHR) DNA vaccination induces high TSHR antibody levels in BALB/c mice housed in a conventional facility. However, under pathogen-free conditions, we observed a Th1 cellular response to TSHR antigen characterized by interferon gamma (IFN gamma) production. In the present study we investigated the effect on TSHR DNA vaccination of diverting the cytokine milieu away from Th1 using 1) IFN gamma knockout BALB/c mice, and 2) wild-type mice covaccinated with DNA for the TSHR and for IFN gamma/receptor-Fc protein that prevents IFN gamma from binding to its receptor. Neither approach enhanced TSHR antibody levels, although splenocyte IFN gamma production in response to TSHR antigen was absent (IFN gamma knockouts) or reduced (IFN gamma receptor-Fc). Moreover, production of IL-2, another Th1 cytokine, but not Th2 cytokines, indicated that neither strategy overcame the Th1 bias of im DNA vaccination. Importantly, splenocyte production of IFN gamma and IL-2 provides a sensitive detection system for TSHR-specific T cells. Unexpectedly, higher TSHR antibody levels developed in rare mice. High titer animals had TSHR-specific responses of both Th2 and Th1 types, whereas low titer animals had Th1-restricted TSHR responses. The heterogeneity of responses induced by TSHR DNA vaccination in mice may provide insight into the titers and IgG subclasses of spontaneous autoantibodies in humans. PMID- 11897672 TI - Substitution of cysteine for a conserved alanine residue in the catalytic center of type II iodothyronine deiodinase alters interaction with reducing cofactor. AB - Human type II iodothyronine deiodinase (D2) catalyzes the activation of T(4) to T(3). The D2 enzyme, like the type I (D1) and type III (D3) deiodinases, contains a selenocysteine (SeC) residue (residue 133 in D2) in the highly conserved catalytic center. Remarkably, all of the D2 proteins cloned so far have an alanine two residue-amino terminal to the SeC, whereas all D1 and D3 proteins contain a cysteine at this position. A cysteine residue in the catalytic center could assist in enzymatic action by providing a nucleophilic sulfide or by participating in redox reactions with a cofactor or enzyme residues. We have investigated whether D2 mutants with a cysteine (A131C) or serine (A131S) two residue amino terminal to the SeC are enzymatically active and have characterized these mutants with regard to substrate affinity, reducing cofactor interaction and inhibitor profile. COS cells were transfected with expression vectors encoding wild-type (wt) D2, D2 A131C, or D2 A131S proteins. Kinetic analysis was performed on homogenates with dithiothreitol (DTT) as reducing cofactor. The D2 A131C and A131S mutants displayed similar Michaelis-Menten constant values for T(4) (5 nM) and reverse T(3) (9 nM) as the wt D2 enzyme. The limiting Michaelis Menten constant for DTT of the D2 A131C enzyme was 3-fold lower than that of the wt D2 enzyme. The wt and mutant D2 enzymes are essentially insensitive to propylthiouracil [concentration inhibiting 50% of activity (IC(50)) > 2 mM] in the presence of 20 mM DTT, but when tested in the presence of 0.2 mM DTT the IC(50) value for propylthiouracil is reduced to about 0.1 mM. During incubations of intact COS cells expressing wt D2, D2 A131C, or D2 A131S, addition of increasing amounts of unlabeled T(4) resulted in the saturation of [(125)I]T(4) deiodination, as reflected in a decrease of [(125)I]T(3) release into the medium. Saturation first appeared at medium T(4) concentrations between 1 and 10 nM. IN CONCLUSION: substitution of cysteine for a conserved alanine residue in the catalytic center of the D2 protein does not inactivate the enzyme in vitro and in situ, but rather improves the interaction with the reducing cofactor DTT in vitro. PMID- 11897673 TI - Molecular regulation of the IGF-binding protein-4 protease system in human fibroblasts: identification of a novel inducible inhibitor. AB - The IGF-binding protein-4 (IGFBP-4) protease system is an important regulator of local IGF bioavailability and cell growth. Recently, the IGF-dependent IGFBP-4 protease secreted by cultured human fibroblasts was identified as pregnancy associated plasma protein A (PAPP-A). In pregnancy serum, PAPP-A circulates as a disulfide-bound complex with the precursor form of major basic protein (pro-MBP), and in this complex PAPP-A's proteolytic activity is not evident. In this study we analyzed the IGFBP-4 protease system in normal human fibroblasts to determine regulation outside of pregnancy. Treatment with the phorbol ester tumor promoter, beta-phorbol 12,13-didecanoate (beta-PDD), resulted in time-dependent inhibition of the IGF-dependent IGFBP-4 protease activity in cell-conditioned medium, which was evident at 6 h and complete by 24 h. PAPP-A mRNA was constitutively expressed in control cells, and levels were decreased only after 24 h of beta-PDD treatment. Secretion of PAPP-A protein into conditioned medium did not change with beta-PDD treatment. On the other hand, pro-MBP mRNA was undetectable in control human fibroblasts, and treatment with beta-PDD induced pro-MBP mRNA and protein expression within 6 h. beta-PDD-induced pro-MBP mRNA expression and protease inhibition were blocked with an inhibitor of RNA synthesis, actinomycin D. Actinomycin D had no effect on PAPP-A mRNA levels in the absence or presence of beta-PDD. Similarly, transformation of human fibroblasts with simian virus 40 large T antigen resulted in the synthesis of pro-MBP mRNA and protein and inhibition of IGFBP-4 protease activity. Coculture of fibroblasts with cells transfected with pro-MBP cDNA resulted in inhibition of IGFBP-4 proteolytic activity without having any effect on PAPP-A synthesis. In summary, phorbol ester tumor promoters and simian virus 40 transformation regulate IGFBP-4 proteolysis in human fibroblasts through induction of a novel inhibitor of PAPP-A, pro-MBP. These findings expand our understanding of the IGFBP-4 protease system and suggest an additional level of local cell growth control. PMID- 11897674 TI - Igf-I inhibits spontaneous apoptosis in human granulocytes. AB - Granulocytes are key cells in inflammatory processes that are recruited to sites of inflammation by chemoattractants such as IL-8 produced by neutrophils and monocytes. Programmed cell death (apoptosis) of granulocytes and subsequent recognition and phagocytosis by macrophages is a crucial mechanism for resolution of inflammation. Because IGF-I is a potent antiapoptotic factor, we addressed the effects of IGF-I on in vitro apoptosis of human peripheral blood granulocytes. We detected 1390 +/- 467 IGF-I receptors with a dissociation constant of 2.3 +/- 0.9 nM on purified granulocytes. Using microscopical analysis, annexin V binding assays to detect relocation of phosphatidylserine to the cell surface, and DNA fragmentation assays, we showed that IGF-I inhibits spontaneous apoptosis of granulocytes in serum-free culture by 32-45%. IGF-I did not modulate the secretion of IL-6, TNF alpha, and IL-8 by granulocytes, but IL-8 secretion by peripheral blood mononuclear cells was enhanced by 40%. These observations indicate that IGF-I may promote granulocyte functions by increasing granulocyte longevity. PMID- 11897675 TI - Sciatic nerve lipoprotein lipase is reduced in streptozotocin-induced diabetes and corrected by insulin. AB - The metabolic abnormalities underlying the cause of diabetic neuropathy have been the subject of much debate. Lipoprotein lipase (LPL) is a 56-kDa enzyme produced by several tissues in the body and has recently been shown in vitro to be expressed in cultured Schwann cells, where it is important in phospholipid synthesis. This suggests a role for LPL in myelin biosynthesis in the peripheral nervous system. The aim of this study was to determine if acute streptozotocin (STZ)-induced diabetes reduces the expression and regulation of sciatic nerve LPL in vivo. Adult Sprague Dawley rats were rendered diabetic via an sc injection of STZ. A decrease in sciatic nerve LPL activity was observed in the STZ-treated rats after just 2 d of diabetes and remained significantly reduced for at least 35 d. The decrease in LPL activity coincided temporally with a drop in motor nerve conduction velocity. Treatment with insulin for 4 d showed a normalization of sciatic nerve LPL activity. These results show that STZ-induced diabetes causes a decrease in LPL activity in the sciatic nerve that, as in other tissues, is reversible with insulin treatment. These data may suggest a role for LPL in the pathophysiology of diabetic neuropathy. PMID- 11897676 TI - Characterization of new selective somatostatin receptor subtype-2 (sst2) antagonists, BIM-23627 and BIM-23454. Effects of BIM-23627 on GH release in anesthetized male rats after short-term high-dose dexamethasone treatment. AB - We here report a pharmacological characterization of two new somatostatin (SS) receptor subtype-2 (sst2) selective antagonists by evaluating their GH-releasing activity when administered, by different routes, in anesthetized adult rats and in freely moving 10-d-old rats. Moreover, we describe the effect of these SS antagonists on the GH response to GHRH after short-term high-dose dexamethasone (DEX) treatment in young male rats. BIM-23454 and BIM-23627, given iv, were able to counteract the SS-induced inhibition of GH secretion occurring after urethane anesthesia in a dose-dependent manner. In DEX-treated animals, the GH response to GHRH was partially blunted (5-min peak values, 270 +/- 50 ng/ml in saline-treated vs. 160 +/- 10 ng/ml in DEX-treated, P < 0.05); however, the simultaneous administration of BIM-23627 (0.2 mg/kg, iv) restored higher amplitude GH pulse, leading to a significantly higher overall mean GH response (area under the curve, 4200 +/- 120 ng/ml/30 min vs. 2800 +/- 100 ng/ml/30 min after GHRH alone; P < 0.05). The SS antagonists showed a reduced GH-releasing effect when administered sc or ip, likely attributable to decreased bioavailability, as compared with the iv route. SS antagonist administration also increased plasma glucagon, insulin, and glucose levels. Based on prior reports that sst2 tonically suppresses glucagon secretion, the antagonist most likely increased glucagon secretion from the pancreatic alpha-cells, with resultant increases in plasma glucose and then insulin. PMID- 11897677 TI - Double-stranded RNA cooperates with interferon-gamma and IL-1 beta to induce both chemokine expression and nuclear factor-kappa B-dependent apoptosis in pancreatic beta-cells: potential mechanisms for viral-induced insulitis and beta-cell death in type 1 diabetes mellitus. AB - Viral infections may trigger the autoimmune assault leading to type 1 diabetes mellitus. Double-stranded RNA (dsRNA) is produced by many viruses during their replicative cycle. The dsRNA, tested as synthetic poly(IC) (PIC), in synergism with the proinflammatory cytokines interferon-gamma (IFN-gamma) and/or IL-1 beta, results in nitric oxide production, Fas expression, beta-cell dysfunction, and death. Activation of the transcription nuclear factor-kappa B (NF-kappa B) is required for PIC-induced inducible nitric oxide synthase expression in beta cells, and we hypothesized that this transcription factor may also participate in PIC-induced Fas expression and beta-cell apoptosis. This hypothesis, and the possibility that PIC induces expression of additional chemokines and cytokines (previously reported as NF-kappa B dependent) in pancreatic beta-cells, was investigated in the present study. We observed that the PIC-responsive region in the Fas promoter is located between nucleotides -223 and -54. Site-directed mutations at the NF-kappa B and CCAAT/enhancer binding protein-binding sites prevented PIC-induced Fas promoter activity. Increased Fas promoter activity was paralleled by enhanced susceptibility of PIC + cytokine-treated beta-cells to apoptosis induced by Fas ligand. beta-Cell infection with the NF-kappa B inhibitor AdI kappa B((SA)2) prevented both necrosis and apoptosis induced by PIC + IL-1 beta or PIC + IFN-gamma. Messenger RNAs for several chemokines and one cytokine were induced by PIC, alone or in combination with IFN-gamma, in pancreatic beta-cells. These included IP-10, interferon-gamma-inducible protein 10, IL-15, macrophage chemoattractant protein-1, fractalkine, and macrophage inflammatory protein-3 alpha. There was not, however, induction of IL-1 beta expression. We propose that dsRNA, generated during a viral infection, may contribute for beta-cell demise by both inducing expression of chemokines and IL 15, putative contributors for the build-up of insulitis, and by synergizing with locally produced cytokines to induce beta-cell apoptosis. Activation of the transcription factor NF-kappa B plays a central role in at least part of the deleterious effects of dsRNA in pancreatic beta-cells. PMID- 11897679 TI - Expression of nitric oxide synthases in rat adrenal zona fasciculata cells. AB - Nitric oxide (NO) synthase (NOS) expression was analyzed in rat adrenal zona fasciculata. Both neuronal NOS and endothelial NOS mRNAs were detected by RT-PCR, immunohistochemistry, and immunoblot analysis. The biochemical characterization of adrenal zona fasciculata NOS enzymatic activity confirmed the presence of a constitutive isoform. In a cell line derived from mouse adrenal cortex, only endothelial NOS expression was detected by both RT-PCR and immunoblot analysis. Nitrate plus nitrite levels in Y1 cell incubation medium were increased in the presence of L-arginine and the calcium ionophore A23187, but not D-arginine, indicating enzymatic activity. Moreover, a low, but significant, conversion of Larginine to L-citrulline, abolished by the NOS inhibitor, N(G)-nitro-L-arginine, was detected in Y1 cells. The effect of L-arginine on pregnenolone production was examined. L-Arginine decreased both basal and ACTH-stimulated pregnenolone production in Y1 cells. The inhibitory effect of L-arginine could be attributed to endogenously generated NO, because it was blocked by N(G)-nitro-L-arginine, and it was mimicked by the addition of a NO donor, diethylenetriamine-NO. An inhibitory effect of NO on pregnenolone production from 22Rhydroxycholesterol and on steroidogenic acute regulatory protein expression was also determined. Taken together, these results suggest that at least part of the adrenal NO could derive from steroidogenic cells and modulate their function. PMID- 11897680 TI - The ubiquitin-proteasome pathway regulates the availability of the GH receptor. AB - GH promotes not only longitudinal growth in children but is active throughout life in protein, fat, and carbohydrate metabolism. The multiple actions of GH start when GH binds to the cell surface-expressed GH receptor. Effectiveness of the hormone depends both on its presence in the circulation and the availability of receptors at the cell surface of target cells. In this study, we examined the role of the ubiquitin-proteasome pathway in regulating GH receptor availability. We show that receptor turnover is rapid, and almost 3-fold prolonged in the internalization-deficient mutant GH receptor (F327A). Using a monovalent GH antagonist, B2036, we could quantify the internalization of the nonactivated receptor. By comparing internalization of the receptor with shedding of the GH binding protein, we show that in Chinese hamster lung cell lines, internalization followed by lysosomal degradation is the major pathway for receptor degradation and that the ubiquitin-proteasome pathway controls this process. Inhibition of endocytosis resulted in a 200% increase in receptor availability at the cell surface at steady state. PMID- 11897681 TI - Pituitary adenylate cyclase-activating polypeptide receptors mediating insulin secretion in rodent pancreatic islets are coupled to adenylate cyclase but not to PLC. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a potentiator of glucose-induced insulin secretion. PACAP binds to a PACAP-specific receptor (PAC1) and to VPAC receptors (VPAC1 and VPAC2), which share high affinity for vasoactive intestinal polypeptide (VIP). In the present study, the molecular expression of PACAP receptor isoforms and the signaling pathways involved in the insulin secretory effect of PACAP were investigated in isolated rat and mouse pancreatic islets. mRNA encoding PAC1-short, -hop, and -very short variants, as well as VPAC1 and VPAC2, were expressed in pancreatic islets. PACAP and VIP were equipotent in potentiating glucose-induced insulin release. Both peptides were also equipotent in increasing cAMP production, but PACAP was more efficient than VIP. Unlike carbachol, PACAP and VIP had no effect on inositol phosphate production. In the PAC1-deficient mouse, the insulinotropic effect of PACAP was reduced, and its differential effect on cAMP production was abolished, whereas the effects of VIP remained unchanged. These results clearly show that the insulinotropic effect of PACAP involved both VPAC and PAC1. The PAC1 variants expressed in rat and mouse pancreatic islets seem to be coupled to adenylate cyclase but not to PLC. PMID- 11897682 TI - Connective tissue growth factor/IGF-binding protein-related protein-2 is a mediator in the induction of fibronectin by advanced glycosylation end-products in human dermal fibroblasts. AB - Expansion of extracellular matrix with fibrosis occurs in many tissues, including skin, as part of the end-organ complications in diabetes. Advanced glycosylation end-products (AGEs) have been implicated as a pathogenic factor in diabetic tissue fibrosis. Connective tissue growth factor (CTGF), also known as IGF binding protein-related protein-2, induces extracellular matrix. We have recently shown that CTGF mRNA and protein are up-regulated by AGE treatment of cultured human dermal fibroblasts. The aim of this study was to determine whether CTGF is an autocrine mediator in the induction of fibronectin (FN) by AGE. Primary cultures of nonfetal human dermal fibroblasts in confluent monolayer were treated with synthesized soluble AGE BSA, 0-200 microg/ml. Analysis of mRNA, by quantitative real-time RT-PCR and conditioned media from treated cultures, showed that FN mRNA was increased by approximately 4-fold at 48 h, and FN protein levels by Western immunoblot and FN ELISA were doubled, compared with control. In the same system, added recombinant human CTGF (0-500 ng/ml) induced FN mRNA and protein levels dose dependently and in a rapid time course. To test whether AGE BSA acts through cell-derived CTGF to induce FN, a CTGF neutralizing antibody was shown to significantly attenuate, but not fully inhibit, the AGE induction of FN mRNA. A pan-specific PKC inhibitor, GF109203X, at 0.2 microM, inhibited the induction of FN mRNA by AGE BSA. Although the same inhibitor did not significantly affect the induction of CTGF mRNA by AGE, it blocked the induction of FN mRNA by recombinant human CTGF. In summary, the induction of FN by AGE is partly mediated by the AGE-induced up-regulation of cell-derived CTGF and is dependent on PKC activity. These results have potential implications for the expansion of extracellular matrix in diabetes mellitus by advanced glycosylation end products. PMID- 11897683 TI - Disruption of the D2 dopamine receptor alters GH and IGF-I secretion and causes dwarfism in male mice. AB - We determined the consequences of the loss of D2 receptors (D2R) on the GH-IGF-I axis using mice deficient in functional dopamine D2 receptors by targeted mutagenesis (D2R(-/-)). Body weights were similar at birth, but somatic growth was less in male D2R(-/-) mice from 1-8 months of age and in D2R(-/-) females during the first 2 months. The rate of skeletal maturation, as indexed by femur length, and the weight of the liver and white adipose tissue were decreased in knockout male mice even though food intake was not altered. The serum GH concentration was significantly decreased during the first 2 months in knockout female and male mice, and IGF-I and IGF-binding protein-3 levels were lower in knockout mice. PRL was significantly higher in knockout mice, and females attained higher levels than males. Pituitaries from adult knockout mice had impaired basal GH release and a lower response to GHRH in vitro. We propose that the D2R participates in GHRH/GH release in the first month of life. In accordance, the D2R antagonist sulpiride lowered GH levels in 1-month-old wild type mice. Our results indicate that lack of D2R alters the GHRH-GH-IGF-I axis, and impairs body growth and the somatotrope population. PMID- 11897684 TI - Transcriptional activation of human CYP17 in H295R adrenocortical cells depends on complex formation among p54(nrb)/NonO, protein-associated splicing factor, and SF-1, a complex that also participates in repression of transcription. AB - The first 57 bp upstream of the transcription initiation site of the human CYP17 (hCYP17) gene are essential for both basal and cAMP-dependent transcription. EMSA carried out by incubating H295R adrenocortical cell nuclear extracts with radiolabeled -57/-38 probe from the hCYP17 promoter showed the formation of three DNA-protein complexes. The fastest complex contained steroidogenic factor (SF-1) and p54(nrb)/NonO, the intermediate complex contained p54(nrb)/NonO and polypyrimidine tract-binding protein-associated splicing factor (PSF), and the slowest complex contained an SF-1/PSF/p54(nrb)/NonO complex. (Bu)(2)cAMP treatment resulted in a cAMP-inducible increase in the binding intensity of only the upper complex and also activated hCYP17 gene transcription. SF-1 coimmunoprecipitated with p54(nrb)/NonO, indicating direct interaction between these proteins. Functional assays revealed that PSF represses basal transcription. Further, the repression of hCYP17 promoter-reporter construct luciferase activity resulted from PSF interacting with the corepressor mSin3A. Trichostatin A attenuated the inhibition of basal transcription, suggesting that a histone deacetylase interacts with the SF-1/PSF/p54(nrb)/NonO/mSin3A complex. Our studies lend support to the idea that the balance between transcriptional activation and repression is essential in the control of adrenocortical steroid hormone biosynthesis. PMID- 11897685 TI - Differential recognition of a tyrosine-dependent signal in the basolateral and endocytic pathways of thyroid epithelial cells. AB - Trafficking of receptors is of crucial importance for the physiology of most exocrine and endocrine organs. It is not known yet if the same mechanisms are used for sorting in the exocytic and endocytic pathways in the different epithelial tissues. In this work, we have used a deletion mutant of the human neurotrophin receptor p75(hNTR) that is normally localized on the apical membrane when expressed in Madin-Darby canine kidney cells. This internal 57-amino acid deletion of the cytoplasmic tail leads to a relocation of the protein from the apical to the basolateral membrane and to rapid and efficient endocytosis. These events are mediated by a signal localized within 9 amino acids of the mutated cytoplasmic tail that is strictly dependent on a tyrosine residue (Tyr-308). We have analyzed the basolateral sorting efficiency and endocytic capacity of this signal in Fischer rat thyroid (FRT) cells, in which basolateral and endocytic determinants have not yet been identified. We found that this targeting signal can mediate efficient transport to the basolateral membrane also in FRT cells with similar tyrosine dependence as in MDCK cells. In contrast to MDCK cells, this Tyr-based signal was not able to mediate coated pits localization and endocytosis in FRT cells. These data represent the first characterization of basolateral/endocytic signals in thyroid epithelial cells. Furthermore, our results indicate that requirements for tyrosine-dependent basolateral sorting signals are conserved among cell lines from different tissues but that the recognition of the colinear endocytic signal is tissue specific. PMID- 11897686 TI - Aggregation of human wild-type and H27A-prolactin in cells and in solution: roles of Zn(2+), Cu(2+), and pH. AB - Aggregation of hormones is an important step in the formation of secretory granules that results in concentration of hormones. In transfected AtT20 cells, but not COS cells, Lubrol-insoluble aggregates of human prolactin (PRL) accumulated within 30 min after synthesis. Aggregation in AtT20 cells was reduced by incubation with 30 microM chloroquine, which neutralizes intracellular compartments, and was slowed by incubation with diethyldithiocarbamate, which chelates Cu(2+) and Zn(2+). H27A-PRL aggregated in AtT20 cells as well as wild type PRL, indicating that a high affinity Zn(2+)-binding site is not necessary. In solution, purified recombinant human PRL was precipitated by 20 microM Cu(2+) or Zn(2+). In solution without polyethylene glycol there was no precipitation with acidic pH alone, precipitation with Zn(2+) was most effective at neutral pH, and the ratio of Zn(2+) to PRL was greater than 1 in the precipitate. In solution with polyethylene glycol, precipitation occurred with acidic pH, precipitation with Zn(2+) occurred effectively at acidic pH, and the ratio of Zn(2+) to PRL was less than 1. The aggregates obtained in polyethylene glycol are therefore better models for aggregates in cells. Unlike human PRL, aggregation of rat PRL has been shown to occur at neutral pH in cells and in solution, and therefore these two similar proteins form aggregates that are the cores of secretory granules in ways that are not completely identical. PMID- 11897687 TI - Regulation of RGS3 and RGS10 palmitoylation by GnRH. AB - Regulators of G protein signaling (RGS) play a pivotal role in cellular signal transduction. RGS3 or RGS10 were overexpressed in GGH(3) cells [GH(3) cells stably expressing the GnRH receptor (GnRHR)]. Responsiveness to a GnRH agonist was assessed because RGS proteins attenuate production of inositol phosphates (IP) and/or cAMP, molecules believed to be involved in GnRH signaling. In addition, site-directed mutagenesis of a potentially palmitoylated Cys(60) residue of RGS10 was used to assess the significance of this site. We observed maximum inhibition of GnRH-stimulated IP responses by RGS3 and by the conserved domain of RGS10 at both 48 and 72 h after transfection, indicating their involvement in G(q)alpha mediated signaling. Significantly diminished cAMP production was observed at all times when cells overexpressed the conserved domain of RGS10; no effect was observed with RGS3 on G(s)alpha-mediated signaling. Palmitic acid incorporation into RGS3 was dependent on agonist occupancy of GnRHR, whereas palmitoylation of RGS10 was constitutive. Mutation of the conserved Cys(60) residue of RGS10 obviated its negative regulatory action on GnRH-stimulated responses, indicating that this site is crucial for its activity on this system. This study is the first demonstration of a role for palmitoylation of this conserved Cys(60) in mammalian G protein signaling. PMID- 11897688 TI - A composite hormone response element regulates transcription of the rat GHRH receptor gene. AB - To further elucidate the molecular mechanisms underlying the transcriptional regulation of the GHRH receptor (GHRH-R) gene, hormonal regulation of the promoter activity of this gene was examined. An approximately 3-kb genomic fragment spanning the promoter region of the gene was sequenced and the transcription start site was determined by RT-PCR and RNase protection assay. A major start site was localized at -105 (relative to the translation initiation codon, ATG), and a pit-1 binding sequence characteristic of pituitary specific genes was found at -155 to -146. Deletion and mutation studies demonstrated this site to be functional. In the presence of dexamethasone, the GHRH-R promoter (from -2935 to -11) directed luciferase expression in MtT-S cells, a somatotropic cell line, but not in the PC12 cells that normally do not express GHRH-R. While T(3), all trans-RA, and 9cis-RA alone weakly enhanced the reporter gene expression, each of these substances was found to act as a synergistic enhancer in the presence of dexamethasone. Additional deletion and mutation analyses demonstrated a functional RA response element at -1090 to -1074. Two functional glucocorticoid response elements and a T(3) response element were found in an 80 bp 5'-flanking sequence of the pit-1 site. Interestingly, it is suggested that the 6-bp half-site AGGACA (from -209 to -204) functions as a 3'-half-site of T(3) response element as well as a 5'-half-site of one of the glucocorticoid response elements. PMID- 11897689 TI - Identification of a potential receptor for both peptide histidine isoleucine and peptide histidine valine. AB - Peptide histidine isoleucine (PHI), peptide histidine valine (PHV), and vasoactive intestinal polypeptide (VIP) are cosynthesized from the same precursor and share high levels of structural similarities with overlapping biological functions. In this study, the first PHI/PHV receptor was isolated and characterized in goldfish. To study this receptor using homologous peptides, we have also characterized the goldfish prepro-PHI/VIP, and, surprisingly, a shorter transcript lacking the VIP coding region was isolated. A PHI/VIP precursor without the VIP coding sequence has never before been reported. Initial functional expression of the PHI/PHV receptor in Chinese hamster ovary cells revealed that it could be activated by human PHV [50% effective concentration (EC(50)): 43 nM] and to a lesser extent human PHI (EC(50): 133 nM) and helodermin (EC(50): 166 nM) but not fish and mammalian pituitary adenylate cyclase activating polypeptides and VIPs. Subsequent studies indicated that, similar to the pituitary adenylate cyclase-activating polypeptide receptors (PAC1-R, VPAC1 R, and VPAC2-R), the receptor isolated in this study is able to interact with goldfish PHI and its C-terminally extended form, PHV with EC(50) values 93 and 43 nM, respectively. Northern blot and RT-PCR/Southern blot analyses revealed that the PHI/VIP gene is expressed in the intestine, brain, and gall bladder and the PHI/PHV receptor gene is primarily expressed in the pituitary and to a lesser extend in the intestine and gall bladder, suggesting that PHI/PHV may play a role, notably in the regulation of pituitary function. In conclusion, our results demonstrate for the first time the existence of a PHI/PHV receptor, indicating that the functions of PHI and PHV could be mediated by their own receptor in addition to VIP receptors. PMID- 11897690 TI - Evidence that brain-derived neurotrophic factor acts as an autocrine factor on pituitary melanotrope cells of Xenopus laevis. AB - We have investigated the physiological regulation and functional significance of brain-derived neurotrophic factor (BDNF) in the endocrine melanotrope cells of the pituitary pars intermedia of the amphibian Xenopus laevis, which can adapt its skin color to the light intensity of its environment. In black-adapted animals, melanotrope cells produce and release alpha-melanophore-stimulating hormone (alpha-MSH). In white-adapted animals, the activity of melanotrope cells is inhibited by neuronal input. Using Western blotting and immunocytochemistry at the light and electron microscopical level, we have detected both the BDNF precursor and the mature BDNF protein in Xenopus melanotrope cells. In situ hybridization and RT-PCR revealed the presence of BDNF mRNA in the pituitary pars intermedia, indicating that BDNF is synthesized in the melanotropes. Real-time quantitative RT-PCR showed that levels of BDNF mRNA in melanotrope cells are about 25 times higher in black- than in white-adapted animals. Although there is no difference in the amount of stored mature BDNF, the amount of BDNF precursor protein is 3.5 times higher in melanotropes of black-adapted animals than in those of white-adapted animals. These data indicate that BDNF mRNA expression and BDNF biosynthesis are up-regulated in active melanotrope cells. Because immunoelectron microscopy showed that BDNF is located in melanotrope secretory granules, BDNF is probably coreleased with alpha-MSH via the regulated secretory pathway. Superfusion and (3)H-amino acid incorporation studies demonstrated that BDNF stimulates the release of alpha-MSH and the biosynthesis of its precursor protein, POMC. Our results provide evidence that BDNF regulates the activity of Xenopus melanotrope cells in an autocrine fashion. PMID- 11897691 TI - Steroid receptor coactivator-1 deficiency causes variable alterations in the modulation of T(3)-regulated transcription of genes in vivo. AB - Thyroid hormone exerts its biological effect by binding to a TR. Both liganded and unliganded TRs regulate the transcription of T(3)-responsive genes. Cofactors with activating or repressing function modulate the transcriptional regulation by TRs. We showed that steroid receptor coactivator 1 (SRC-1)-deficient mice (SRC-1( /-)) exhibit partial resistance to thyroid hormone at the level of the pituitary thyrotrophs. To determine whether SRC-1 deficiency affects globally T(3) dependent transcriptional regulation, we studied the effects of thyroid hormone deprivation and replacement on the expression of several genes in different tissues of SRC-1(-/-) and wild-type mice (SRC-1(+/+)). Thyroid hormone deficiency was induced by a low iodine diet (LoI) supplemented with propylthiouracil (PTU) for 2 wk. L-T(3) was injected ip for the last 4 d in one group (PTU+T(3) group), and another group (PTU group) received only vehicle. Levels of mRNAs for T(3) responsive genes were determined by Northern blotting: GH and TSH beta in pituitary; type 1 iodothyronine 5'-deiodinase, spot 14 (S14), and malic enzyme in liver; and sarcoplasmic reticulum calcium adenosine triphosphatase 2 and myosin heavy chain alpha and beta in heart. Serum parameters, TSH, total cholesterol, creatine kinase, and alkaline phosphatase (AP), were also measured. Hypothyroidism produced a comparable increase in TSH beta mRNA in both genotypes, but its suppression by L-T(3) was attenuated in SRC-1(-/-) mice. In contrast, hypothyroidism failed to reduce S14 mRNA levels in SRC-1(-/-) mice. As a consequence, the response to L-T(3) was not observed in these mice. SRC-1 deficiency had no effect on the expression of the rest of the T(3)-responsive genes examined. Of the four serum parameters, the T(3)-mediated decrease in TSH and changes in AP were attenuated in SRC-1(-/-) mice. We conclude that SRC-1 deficiency altered the expression of only some of the T(3)-responsive genes. SRC 1 appears to be involved not only in transcriptional activation by liganded TRs, but also in the suppression by liganded or unliganded TRs. Some of the effects of SRC-1 may be TR isoform specific. PMID- 11897692 TI - Ultradian rhythmicity of ghrelin secretion in relation with GH, feeding behavior, and sleep-wake patterns in rats. AB - Ghrelin, an endogenous ligand for the GHS receptor, stimulates GH secretion and gastrointestinal motility and has orexigenic effects. In this study, the relationships between ghrelin, GH secretion, feeding behavior, and sleep-wake patterns were investigated in adult male rats. The half-life of exogenous ghrelin (10 microg i.v.) in plasma was about 30 min. Repeated administration of ghrelin at 3- to 4-h intervals (one during lights-on and two during lights-off periods) increased GH release and feeding activity, and decreased rapid eye movement sleep duration. Endogenous plasma ghrelin levels exhibited pulsatile variations that were smaller and less regular compared with those of GH. No significant correlation between GH and ghrelin circulating levels was found, although mean interpeak intervals and pulse frequencies were close for the two hormones. In contrast, ghrelin pulse variations were correlated with food intake episodes in the lights off period, and plasma ghrelin concentrations decreased by 26% in the 20 min following the end of the food intake periods. A positive correlation between ghrelin levels and active wake was found during the first 3 h of the dark period only. In conclusion, ghrelin, in addition to affecting GH secretion, gastrointestinal motility, and feeding activity, also modifies sleep-wake patterns. However, a direct action of ghrelin per se or the indirect effects of feeding (and all of its attendant metabolic sequelae) on sleep cannot be differentiated. Moreover, ghrelin secretion is pulsatile and directly related to feeding behavior only. PMID- 11897693 TI - Decrements in nuclear glucocorticoid receptor (GR) protein levels and DNA binding in aged rat hippocampus. AB - Hippocampal glucocorticoid receptors (GRs) are believed to play a major role in age-related cognitive decline and cellular vulnerability. It has been proposed that these receptors mediate damaging effects of elevated glucocorticoid secretion on cellular function. In the present report we present evidence that intracellular trafficking of the GR is impaired with hippocampal aging, manifest as decreased nuclear translocation and deficient DNA binding. We also present evidence that chaperone proteins responsible for GR nuclear translocation are decreased with hippocampal aging. Age-related nuclear GR decreases are not observed in hypothalamus, indicating regional specificity of trafficking deficits. Aging did not affect nuclear or cytosolic MR levels. These data suggest that GR signaling is diminished, rather than enhanced, during hippocampal aging. Diminished GR signaling capacity may attenuate the beneficial effects of glucocorticoids on hippocampal regulation of memory and stress integration. PMID- 11897695 TI - Targeted deletion of the PRL receptor: effects on islet development, insulin production, and glucose tolerance. AB - PRL and placental lactogen (PL) stimulate beta-cell proliferation and insulin gene transcription in isolated islets and rat insulinoma cells, but the roles of the lactogenic hormones in islet development and insulin production in vivo remain unclear. To clarify the roles of the lactogens in pancreatic development and function, we measured islet density (number of islets/cm(2)) and mean islet size, beta-cell mass, pancreatic insulin mRNA levels, islet insulin content, and the insulin secretory response to glucose in an experimental model of lactogen resistance: the PRL receptor (PRLR)-deficient mouse. We then measured plasma glucose concentrations after ip injections of glucose or insulin. Compared with wild-type littermates, PRLR-deficient mice had 26-42% reductions (P < 0.01) in islet density and beta-cell mass. The reductions in islet density and beta-cell mass were noted as early as 3 wk of age and persisted through 8 months of age and were observed in both male and female mice. Pancreatic islets of PRLR-deficient mice were smaller than those of wild-type mice at weaning but not in adulthood. Pancreatic insulin mRNA levels were 20-30% lower (P < 0.05) in adult PRLR deficient mice than in wild-type mice, and the insulin content of isolated islets was reduced by 16-25%. The insulin secretory response to ip glucose was blunted in PRLR-deficient males in vivo (P < 0.05) and in isolated islets of PRLR deficient females and males in vitro (P < 0.01). Fasting blood glucose concentrations in PRLR-deficient mice were normal, but glucose levels after an ip glucose load were 10-20% higher (P < 0.02) than those in wild-type mice. On the other hand, the glucose response to ip insulin was normal. Our observations establish a physiologic role for lactogens in islet development and function. PMID- 11897694 TI - Induction of uncoupling protein 2 mRNA in beta-cells is stimulated by oxidation of fatty acids but not by nutrient oversupply. AB - We tested for regulation of uncoupling protein 2 (UCP-2) in beta-cells in response to fatty acids and glucose. A 48-h culture with oleate (0.2 mM) at 5.5 or 11 mM glucose increased UCP-2 mRNA by 30-60% in INS-1 cells and in rat pancreatic islets. In contrast, oleate was ineffective after coculture at 27 mM glucose, P < 0.05 for difference 5.5 vs. 27 mM glucose. Also, culture with palmitate (0.1 mM) stimulated UCP-2 expression at 5.5 and 11 mM, but not at 27 mM glucose. Glucose per se failed to affect UCP-2 mRNA. Oxidation of [1-(14)C] oleate was increased by culture with oleate; however, this increase was attenuated by glucose during coculture, P < 0.05 for coculture at 5.5 vs. 27 mM glucose. Culture with aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside, an activator of AMP-activated protein kinase, decreased cellular triglycerides, increased postculture [1-(14)C] oleate oxidation, and increased UCP-2 mRNA. Etomoxir, an inhibitor of carnitine palmitoyltransferase I, decreased the oleate induced increase in UCP-2 mRNA. Rosiglitazone, a peroxisome proliferator activated receptor gamma ligand, affected neither UCP-2 mRNA nor [1-(14)C] oleate oxidation. Antioxidants (vitamin E and sodium selenite) did not affect oleate induced UCP-2 mRNA. We conclude that: 1) UCP-2 mRNA is induced by fatty acid oxidation in beta-cells; and 2) glucose exerts a modulating effect that is coupled to inhibition of fatty acid oxidation PMID- 11897696 TI - Proteasome implication in phorbol ester- and GnRH-induced selective down regulation of PKC (alpha, epsilon, zeta) in alpha T(3)-1 and L beta T(2) gonadotrope cell lines. AB - We investigated mechanisms underlying selective down-modulation of PKC isoforms (alpha, epsilon, zeta): 1) during 12-O-tetradecanoyl-phorbol-13 acetate (TPA) (10(-7) M) or GnRH (10(-7) M) desensitization conditions (2- to 6-h treatments) in two gonadotrope cell lines (alpha T(3)-1, L beta T(2)) and 2) in primary pituitary cell cultures from male rats during long-term phorbol ester administration. We demonstrated that, as in alpha T(3)-1 cells, in a more differentiated gonadotrope cell line L beta T(2) the GnRH-receptor coupling (PLC, PLA2, PLD) generated second messengers essential for PKCs activation; the characterized isoforms (alpha, beta II, delta, epsilon, zeta) were selectively and differentially down-regulated by TPA (alpha, beta II, delta, epsilon) or GnRH (delta, epsilon). In whole cell lysates, proteasome inhibitors (proteasome inhibitor I and II, Lactacystin, beta-Lactone, Calpain inhibitor I) prevented in both gonadotrope cell lines the TPA-induced depletion of PKC alpha, epsilon, and the GnRH-elicited PKC epsilon down-regulation; they counteracted in mixed pituitary cell cultures as well, the TPA-evoked PKC alpha, epsilon depletion. In contrast, the inhibitors of calpain(s) and lysosomal proteases (Calpeptin, E64d, Calpain inhibitor II, and PD150606), were ineffective. As shown in alpha T(3)-1 subcellular fractions, proteasome abrogation did not affect membrane translocation of TPA- and GnRH- target isoforms (alpha, epsilon) but, preventing their degradation, favored enzyme accumulation to the membrane compartment. Proteolysis processing of PKCs may be dependent upon their phosphorylated state and/or catalytic activity. Inhibition of PKC catalytic activity (GF109203X, Go6976), selectively prevented the TPA-evoked PKC alpha depletion in both mixed pituitary cells and alpha T(3)-1 gonadotropes; in alpha T(3)-1 subcellular fractions, PKC alpha inactivation overcame the TPA-evoked isoenzyme degradation by inducing a pronounced membrane accumulation of the isoform without affecting its membrane relocalization. Thus, the proteasome system by adjusting PKC cellular levels, may represent a regulatory proteolytic pathway implicated in the adaptive mechanisms of the time dependent cell responses. PMID- 11897697 TI - Neuron-specific expression in vivo by defined transcription regulatory elements of the GnRH gene. AB - The GnRH-expressing neurons are the ultimate regulator of reproductive function. GnRH gene expression is limited to this small population of neurons in the hypothalamus. Transfections using 3 kb of the rat or mouse 5'-regulatory region provide specific gene expression in the hypothalamic cell line GT1-7. The combination of two elements, a 300-bp enhancer and a 173-bp promoter, recapitulates specificity in GT1-7 cells. It was not known whether these elements could specifically target gene expression throughout development in the whole animal. We demonstrate that the 3-kb rat GnRH regulatory region provides a higher degree of specificity than the equivalent mouse sequence in a mouse hypothalamic cell line. Moreover, combination of the enhancer and the promoter of the rat gene targets expression to GnRH neurons in transgenic mice in a developmentally appropriate manner. Transgene expression is regulated by activin A, a known activator of GnRH gene expression. In contrast, the enhancer on a heterologous promoter produces inappropriate expression in vivo. We conclude that the enhancer and promoter regions of the rat GnRH gene are necessary for targeted expression to hypothalamic neurons and are sufficient to confer regulated, cell type specific expression to a reporter gene in vivo. PMID- 11897698 TI - Colony-stimulating factor 1 regulation of neuroendocrine pathways that control gonadal function in mice. AB - Colony stimulating factor 1 (CSF-1) is the primary regulator of cells of the mononuclear phagocytic lineage. Consequently mice lacking CSF-1 (Csf1(op)/Csf1(op)) have depleted populations of macrophages in many tissues. In addition, both sexes have reduced fertility with females having extended estrus cycles and poor ovulation rates, whereas males have low circulating LH and T. In this study, we show that puberty was significantly delayed in Csf1(op)/Csf1(op) females compared with control littermates. Restoration of circulating CSF-1 over the first 2 wk of life accelerated puberty, and this treatment until puberty completely corrected the extended estrous cycles. In a standard LH surge induction protocol, Csf1(op)/Csf1(op) females showed diminutive negative and no positive feedback response to E2. These data, together with that from male Csf1(op)/Csf1(op) mice that showed normal release of LH with a GnRH agonist, indicate that the hypothalamus is the site of the primary defect causing fertility problems in CSF-1-deficient mice. In the hypothalamus, microglia are the only CSF-1 receptor-bearing cells, and the recruitment of a full complement these cells is slightly delayed in Csf1(op)/Csf1(op) mice. These data suggest a role for CSF-1 and its target cells, microglia, in establishing the feedback sensitivity to circulating steroid hormones in the hypothalamus of mice. PMID- 11897700 TI - Expression and localization of activin receptors, Smads, and beta glycan to the postnatal rat ovary. AB - Despite understanding the molecular basis of activin/TGF beta and bone morphogenetic protein (BMP) signaling, this study is the first to characterize multiple, sequential elements of these pathways in the ovary concurrently. The expression of activin/BMP receptor, Smad, and beta glycan mRNAs by postnatal rat ovaries were investigated by real-time PCR. Activin/BMP receptors (ActRIA, ActRIB, ActRIIA, and ActRIIB), beta glycan, and Smad 1-8 mRNAs were expressed by the ovary. Activin receptor and Smad 1, 2, 4, 5, and 7 mRNAs declined up to 4 fold between postnatal d 4-8, coinciding with secondary follicle formation. The emergence of antral follicles (postnatal d 12) saw ActRIA, ActRIIB, and Smad 2 mRNA expression return to d 4 levels, whereas ActRIB, ActRIIA, and Smads 1, 4, 5, and 7 remained at lower levels. beta glycan mRNA levels increased 2-fold between d 8 and 12, suggesting expression by the developing theca. Smad 3, 6, and 8 mRNAs were unchanged. Activin receptor and Smad proteins were present in oocytes at all stages of follicular development; granulosa cells of primary-antral follicles, and theca cells. beta glycan protein was present in oocytes, granulosa cells, and theca cells at all stages of folliculogenesis. The colocalization of receptors and Smads supports the notion that activin/TGF beta and BMP signaling pathways are functional in the cellular compartments of the follicle. PMID- 11897701 TI - Production of the chemokines monocyte chemotactic protein-1, regulated on activation normal T cell expressed and secreted protein, growth-related oncogene, and interferon-gamma-inducible protein-10 is induced by the Sendai virus in human and rat testicular cells. AB - Several viruses infect the testis, inducing inflammation, which may lead to infertility. In this study we investigated the production in rat and human testicular cells exposed to the Sendai virus of several chemokines that play a major role in inflammatory processes. Exposure of rat testicular macrophages and Sertoli, Leydig, and peritubular cells to the Sendai virus led to the production of mRNA and protein for monocyte chemotactic protein-1 (MCP-1), regulated on activation normal T cell expressed and secreted protein, growth-related oncogene alpha, and interferon-gamma-inducible protein-10. In rat peritubular cells exposed to the Sendai virus, MCP-1 production was time and dose dependent. In contrast, rat germ cells did not produce these chemokines. Chemokine synthesis was detected in human Leydig cells exposed to the Sendai virus, but not in human total germ cells, suggesting that rats and humans display similar responses in terms of chemokine production. MCP-1, regulated on activation normal T cell expressed and secreted protein, growth-related oncogene-alpha, and interferon gamma-inducible protein-10 have been reported to be chemoattractants for a large variety of leukocytes. The ability of the Sendai virus to induce chemokine production in somatic cells (mostly peritubular and Leydig cells) may therefore increase the recruitment of leukocytes to sites of infection. PMID- 11897702 TI - The angiogenic factor cysteine-rich 61 (CYR61, CCN1) supports vascular smooth muscle cell adhesion and stimulates chemotaxis through integrin alpha(6)beta(1) and cell surface heparan sulfate proteoglycans. AB - Cysteine-rich 61 (CYR61, CCN1) is a heparin-binding, extracellular, matrix associated protein of the cysteine-rich 61/nephroblastoma family, which also includes connective tissue growth factor, nephroblastoma overexpressed, Wnt induced secreted protein-1 (WISP-1), WISP-2, and WISP-3. CYR61 induces angiogenesis in vivo and supports cell adhesion, promotes cell migration, and enhances growth factor-stimulated mitogenesis in fibroblasts and endothelial cells. Although the expression of CYR61 has been observed in arterial walls, its function in vascular smooth muscle cells (VSMCs) has not been examined to date. Here we show that purified CYR61 supports VSMC adhesion in a dose-dependent, saturable manner through integrin alpha(6)beta(1) with an absolute requirement of cell surface heparan sulfate proteoglycans. In addition, CYR61 induces VSMC chemotaxis, but not chemokinesis, through integrin alpha(6)beta(1) and heparan sulfate proteoglycans. Heparin-binding defective CYR61 mutants are unable to support VSMC adhesion but can still induce chemotaxis at a reduced level. Following balloon angioplasty in rat carotid artery, CYR61 protein level is elevated in the media and neointima of the injured vessel by d 4 post angioplasty, peaks from d 7 to 14, and remains high for at least 28 d. These data demonstrate the activities of CYR61 in VSMCs, identify the receptors that mediate its functions, and show that CYR61 is synthesized in arterial smooth muscle walls during proliferative restenosis. Together, these results implicate CYR61 as a novel factor that modulates the responses of VSMCs to vascular injury. PMID- 11897703 TI - Adrenocortical cytochrome b5 expression during fetal development of the rhesus macaque. AB - The developmental expressions of cytochrome b5 (b5), 17 alpha-hydroxylase/17,20 lyase cytochrome P450 (P450c17), and 3 beta-hydroxysteroid dehydrogenase were examined in primate fetal adrenals by immunocytochemistry from 50-160 d gestation. The expression of b5 was evident at 50 d in the developing fetal zone (FZ), but decreased markedly through midgestation, then increased again from 150 d to term. Similar changes in the temporal expression was observed for P450c17. Whereas P450c17 was induced largely in the transitional zone (TZ; outer-most FZ), b5 expression was strongest in FZ cells further from the capsule, although overlap between these regions involved a narrow band of cells beneath the TZ that may represent the developing zona reticularis. Thus, the induction of b5 in the FZ and of P450c17 in the TZ of the fetal adrenal late in gestation coincided temporally with the prepartum rise in dehydroepiandrosterone previously reported. These data are consistent with the proposed role of b5 in supporting 17,20-lyase activity of P450c17. However, the lack of cytochrome b5 and P450c17 expression in the FZ of the developing macaque adrenal cortex for much of the second and third trimesters distinguishes it from the mature zona reticularis seen in adult animals. PMID- 11897704 TI - Effect of GABA on GnRH neurons switches from depolarization to hyperpolarization at puberty in the female mouse. AB - The amino acid gamma-aminobutyric acid (GABA) plays an important role in the regulation of the GnRH neurons. We examined whether GABA depolarizes or hyperpolarizes GnRH neurons over postnatal development using gramicidin, perforated-patch electrophysiology combined with GnRH-LacZ transgenic mice in whom GnRH neurons can be made to fluoresce. The basic membrane properties and GABA responsiveness of GnRH neurons were not altered by transgene expression or fluorescence. Ten of 12 immature GnRH neurons (10-17 d) were depolarized by GABA in a direct and dose-dependent manner that was blocked by a GABA(A) receptor antagonist. In peripubertal GnRH neurons (25-30 d), GABA exerted depolarizing (4/11) as well as hyperpolarizing (5/11) effects on GnRH neurons. In adult female mice, GABA was found to exert exclusively hyperpolarizing actions on GnRH neurons (9/10) that were direct and mediated by the GABA(A) receptor. GABA switched from depolarizing to hyperpolarizing actions around postnatal d 31, the time of vaginal opening. Unidentified preoptic area neurons exhibited predominantly hyperpolarizing responses to GABA at all three postnatal stages. These findings demonstrate that GnRH neurons display an unusually late postnatal switch in their response to GABA. They also provide the first direct evidence that GABA inhibits the electrical activity of postpubertal GnRH neurons. PMID- 11897705 TI - Extracellular Ca(2+)-sensing receptors modulate matrix production and mineralization in chondrogenic RCJ3.1C5.18 cells. AB - Previous studies in chondrogenic RCJ3.1C5.18 (C5.18) cells showed that growth of these cells at high extracellular Ca(2+) concentrations ([Ca(2+)](o)) reduced the expression of markers of early chondrocyte differentiation. These studies addressed whether raising [Ca(2+)](o) accelerates C5.18 cell differentiation and whether Ca(2+) receptors (CaRs) are involved in coupling changes in [Ca(2+)](o) to cellular responses. We found that high [Ca(2+)](o) increased expression of osteopontin (OP), osteonectin, and osteocalcin, all markers of terminal differentiation, in C5.18 cells and increased the production of matrix mineral. Overexpression of wild-type CaR cDNA in C5.18 cells suppressed proteoglycan synthesis and aggrecan RNA, two early differentiation markers, and increased OP expression. The sensitivity of these parameters to changes in [Ca(2+)](o) was significantly increased, as indicated by left-shifted dose-responses. In contrast, stable expression of a signaling-defective CaR mutant (Phe707Trp CaR) in C5.18 cells, presumably through dominant-negative inhibition of endogenous CaRs, blocked the suppression of aggrecan RNA levels and proteoglycan accumulation and the enhancement of OP expression by high [Ca(2+)](o). These data support a role for CaRs in mediating high [Ca(2+)](o)-induced differentiation of C5.18 cells. PMID- 11897706 TI - Localization and regulation of a functional GHRH receptor in the rat renal medulla. AB - To provide information about the kidney GHRH receptor (GHRH-R), we assessed its tissue and cellular localization, defined its pattern of expression in developing and aging rats, and studied the effects of GHRH on the regulation of GHRH-R mRNA levels and receptor internalization. In situ hybridization and ribonuclease protection assay demonstrated that GHRH-R mRNA is restricted to the Henle's loop (HL). GHRH-R mRNA levels were low in the medulla from 3- and 12-d-old male rats, increased significantly in that from 30- to 70-d-old rats, and decreased in that from 12- and 18-month-old animals. Compared with the GHRH-R mRNA profile obtained in the pituitary, these data support the concept of a tissue-specific regulation of GHRH-R. In HL cell cultures from 70-d-old rats, a 4-h incubation with 1-100 nM rat GHRH-(1-29)NH(2) reduced GHRH-R mRNA levels significantly. As anti-GHRH-R- (392-404) immunoreactivity was demonstrated in HL cells, internalization of [N(alpha)-5-carboxyfluoresceinyl-D-Ala(2),Ala(8), Ala(15),Lys(22)]hGHRH-(1 29)NH(2) in a time- and temperature-dependent manner and inhibition of this process by phenyl arsine oxide indicate that desensitization to GHRH involves both GHRH-R internalization and down-regulation of GHRH-R mRNA levels. Localization of a functional GHRH-R in HL and its regulation during development and aging suggest roles associated with cellular proliferation, differentiation, and/or water/electrolyte transport. PMID- 11897707 TI - Nerve growth factor induces the expression of functional FSH receptors in newly formed follicles of the rat ovary. AB - The neurotrophin nerve growth factor (NGF) and its two membrane-anchored receptors are expressed in the developing ovary before the organization of the first primordial follicles. In the absence of NGF, the growth of primordial follicles is retarded, indicating that NGF contributes to facilitating early follicular development. The present experiments were undertaken to determine whether NGF can also be involved in the differentiation process by which ovarian follicles become responsive to gonadotropins. Treatment of 2-d-old rat ovaries in organ culture with NGF increased FSH receptor (FSHR) mRNA within 8 h of exposure. This effect was cAMP-independent but additive to the cAMP-mediated increase in FSHR gene expression induced by either forskolin or vasoactive intestinal peptide, a neurotransmitter previously shown to induce FSHR formation in neonatal rat ovaries. After NGF treatment, the ovary acquired the capacity of responding to FSH with cAMP formation and preantral follicular growth, indicating that exposure to the neurotrophin resulted in the formation of biologically active FSHRs. Quantitative measurement of FSHR mRNA demonstrated that the content of FSHR mRNA is reduced in the ovaries of mice carrying a null mutation of the NGF gene. These results indicate that one of the functions of NGF in the developing ovary is to facilitate the differentiation process by which early growing follicles become gonadotropin-dependent during postnatal life, and that it does so by increasing the synthesis of FSHRs. PMID- 11897708 TI - Caspase-3 is a pivotal mediator of apoptosis during regression of the ovarian corpus luteum. AB - Because caspase-3 is considered a primary executioner of apoptosis and has been implicated as a mediator of luteal regression, we hypothesized that corpora lutea (CL) derived from caspase-3 null mice would exhibit a delayed onset of apoptosis during luteal regression, when compared with CL derived from wild-type (WT) mice. To test this hypothesis, ovulation was synchronized in immature (postpartum d 24 27) WT and caspase-3-deficient female littermates by exogenous gonadotropins. Individual CL were isolated by manual dissection, 30 h after ovulation, and placed in organ culture dishes in the absence of serum and growth factors. At the time of isolation (0 h) and after 24, 48, and 72 h in culture, the CL were removed and assessed for the presence of processed (active) caspase-3 enzyme and for apoptosis by multiple criteria. There was no evidence of active caspase-3 enzyme or apoptosis in either WT or caspase-3-deficient CL before culture. However, CL derived from the WT mice exhibited a time-dependent increase in the level of active caspase-3 and apoptosis during culture. By comparison, CL derived from caspase-3-deficient mice, cultured in parallel, failed to exhibit any detectable active caspase-3 and showed attenuated rates of apoptosis. To extend these findings derived from ex vivo culture experiments, ovaries were collected from WT and caspase-3 null female littermates at 2, 4, or 6 d post ovulation, and the occurrence of apoptosis within the CL was analyzed. Whereas ovaries of WT mice had only residual luteal tissue at d 6 post ovulation, ovaries collected from caspase-3-deficient mice retained many CL, at d 6 post ovulation, that were similar in size to those observed in the early luteal phase of WT mice. Importantly, there was no dramatic increase in apoptosis in CL of caspase-3 deficient mice at any time point examined post ovulation, indicating that the involution process had indeed been delayed. In contrast, the levels of progesterone declined regardless of genotype. These data provide the first direct evidence that caspase-3 is functionally required for apoptosis to proceed normally during luteal regression. However, caspase-3 is not a direct mediator of the decrease in steroidogenesis associated with luteolysis. PMID- 11897709 TI - Exclusive action of transmembrane TNF alpha in adipose tissue leads to reduced adipose mass and local but not systemic insulin resistance. AB - Aberrant TNF alpha expression in adipocytes is a molecular mechanism by which insulin action is modulated in adipose tissue. While this might be a compensatory response to limit adipose expansion, neither the mechanisms underlying this local effect nor its systemic biological consequences have been studied. It is also not clear whether TNF alpha-induced insulin resistance in adipocyte alone is responsible for systemic insulin resistance in the absence of obesity. In a transgenic mouse model deficient in endogenous TNF alpha, we demonstrate that specific expression of the transmembrane TNF alpha (mTNF alpha) in adipocytes leads to decreased whole body adipose mass, and local, but not systemic insulin resistance. These data demonstrate that exclusive action of TNF alpha in adipose tissue strongly inhibits insulin action at this site and leads to reduced adiposity in mice. However, this isolated adipocyte insulin resistance in the context of reduced fat mass and/or the absence of obesity is insufficient to alter systemic glucose homeostasis. PMID- 11897710 TI - Paradoxical regulation of Sp1 transcription factor by glucagon. AB - Insulin is a potent regulator of Sp1 transcription factor. To examine if glucagon, which usually antagonizes insulin, regulates Sp1, we assessed the levels of Sp1 by Western blotting from H-411E cells exposed to glucagon with or without insulin. Glucagon alone (1.5 x 10(-9) to 1.5 x 10(-5) M) stimulated Sp1 accumulation but inhibited insulin's (10,000 microU/ml) stimulatory effect on Sp1. We also assessed the effect of TNF-alpha, wortmannin, a PI3K inhibitor, and cAMP-dependent protein kinase inhibitor on Sp1 accumulation. While TNF-alpha (5 ng/ml) blocked insulin-stimulated Sp1, it failed to block stimulation of Sp1 by glucagon (1.5 x 10(-5) M). Similarly, wortmannin inhibited insulin- but not glucagon-stimulated Sp1, whereas protein kinase inhibitor had an opposite effect. Thus, insulin acts primarily via PI3K, and glucagon apparently stimulates through a cAMP-dependent pathway. Insulin increased the staining intensity of Sp1 seen exclusively in the nuclei of H-411E cells. Sp1 was demonstrable in both nucleus and cytoplasm after glucagon treatment. Finally, as judged by immunoblotting to specific antibody, insulin but not glucagon, stimulated O-glycosylation of Sp1. Thus, unique signaling mechanisms mediate the response of Sp1 to glucagon in the presence or absence of insulin. PMID- 11897711 TI - The mutual regulation of arginine-vasopressin and PTHrP secretion in dissociated supraoptic neurons. AB - PTHrP is detected in the supraoptic nucleus (SON) and paraventricular nucleus. We have recently demonstrated that PTHrP(1-34) is involved in AVP release and synthesis in the SON in vivo and in vitro. PTHrP and AVP, which act on blood vessels, may interact by autocrine and paracrine mechanisms in the central nervous system. The present study was undertaken to determine the mutual regulation of AVP and PTHrP secretion in dissociated magnocellular neurons of the SON. Both AVP and PTHrP existed in the dissociated SON neurons by immunohistochemistry. PTHrP(1-34) stimulated AVP secretion from the cells dose dependently, but PTHrP(7-34) and PTH(1-34) did not. PTHrP(1-34)-stimulated AVP secretion was associated with cAMP generation. PTHrP(1-34)-induced cAMP generation was inhibited by a 100-fold molar excess of PTHrP(7-34) but not by that of PTH(1-34). PTHrP(1-34) also stimulated AVP mRNA expression in the cells. These results are consistent with our previous observations that PTHrP(1-34) is involved in AVP secretion through a receptor distinct from type I PTH/PTHrP receptor. Next, AVP stimulated dose-dependent PTHrP release from the dissociated SON neurons. The AVP-induced PTHrP release was suppressed by both OPC-21268 (V(1a) receptor antagonist) and dP[Thy(Me)(2)]AVP (V(1a)/V(1b) receptor antagonist) but not by OPC-31260 (V(2) receptor antagonist). AVP increased PKC activity dose dependently but not cAMP generation in the SON neurons. The AVP stimulated PTHrP release was blocked by staurosporine (PKC inhibitor), nicardipine (L-type calcium channel blocker) or omega-agatoxin IVA (N type). Furthermore, AVP stimulated PTHrP mRNA expression for 12 h in the SON neurons. These results indicate that AVP caused increases in PTHrP secretion and its mRNA levels through V(1a) and/or V(1b) receptors in the SON neurons. Our observations, taken together, suggest that PTHrP stimulates AVP secretion into the extracellular space of the SON, which in turn leads to further secretion of AVP and PTHrP by an autocrine/paracrine mechanism. PMID- 11897712 TI - Increased islet cell proliferation, decreased apoptosis, and greater vascularization leading to beta-cell hyperplasia in mutant mice lacking insulin. AB - The targeted disruption of the two nonallelic insulin genes in mouse was reported previously to result in intrauterine growth retardation, severe diabetes immediately after suckling, and death within 48 h of birth. We have further used these animals to investigate the morphology and cell biology of the endocrine pancreas in late gestation and at birth when insulin is absent throughout development. Pancreatic beta-cells were identified by detecting the activity of the LacZ gene inserted at the Ins2 locus. A significant increase in the mean area of the islets was found at embryonic d 18.5 (E18.5) and in the newborn in Ins1-/ , Ins2-/- animals compared with Ins1-/-, Ins2+/- and wild-type controls, whereas the blood glucose levels were unaltered. The individual size of the beta-cells in the insulin-deficient fetuses was similar to controls, suggesting that the relative increase in islet size was due to an increase in cell number. Immunohistochemistry for proliferating cell nuclear antigen within the pancreatic ductal epithelium showed no differences in labeling index between insulin deficient and control mice, and no change in the number of beta-cells associated with ducts, but the relative size distribution of the islets was altered so that fewer islets under 5,000 microm(2) and more islets greater than 10,000 microm(2) were present in Ins1-/-, Ins2-/- animals. This suggests that the greater mean islet size seen in insulin-deficient animals represented an enlargement of formed islets and was not associated with an increase in islet neogenesis. The proportional contribution of alpha- and beta-cells to the islets was not altered. This was supported by an increase in the number of cells containing immunoreactive proliferating cell nuclear antigen in both islet alpha- and beta cells at E18.5 in insulin-deficient mice, and a significantly lower incidence of apoptotic cells, as determined by molecular histochemistry using the terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling reaction. The density of blood vessels within sections of whole pancreas, or within islets, was determined by immunohistochemistry for the endothelial cell marker CD31 and was found to be increased 2-fold in insulin-deficient mice compared with controls at E18.5. However, no changes were found in the steady-state expression of mRNAs encoding vascular endothelial growth factor, its receptor Flk-1, IGF-I or -II, the IGF-I and insulin receptors, or insulin receptor substrates-1 or -2 in pancreata from Ins1-/-, Ins2-/- mice compared with Ins1-/-, Ins2+/- controls. Thus, we conclude that the relative hyperplasia of the islets in late gestation in the insulin-deficient mice was due to an increased islet cell proliferation coupled with a reduced apoptosis, which may be related to an increased vascularization of the pancreas. PMID- 11897713 TI - Nicotinamide adenine dinucleotide phosphate-dependent cytosolic T(3) binding protein as a regulator for T(3)-mediated transactivation. AB - Nicotinamide adenine dinucleotide phosphate (NADPH)- dependent cytosolic T(3) binding protein (CTBP) plays a role in the regulation of nuclear transport of T(3) in vitro. However, it is not known whether CTBP regulates the T(3) action. In this study, we examined the effects of CTBP on cellular translocation of T(3) and on transcriptional activation using established CTBP-expressing CHO or GH3 cells. The expression of CTBP increased cellular and nuclear uptake of T(3) in the CTBP-expressing cells. The efflux rate was decreased by induction of CTBP. Efflux from nuclei also inhibited by induction of CTBP. Expression of CTBP suppressed the T(3)-regulated luciferase activity in GH3 cells. Suppression was observed to be related to the expression level of CTBP. T(3) induction of rat GH mRNA was lower in the cells expressing CTBP than that in CTBP-null cells. These results suggest that CTBP regulates the T(3)-induced gene expression, with which an increase in the nuclear content of the T(3) is associated. Because we observed that a part of CTBP could be transported into nuclei and that acceptor protein for CTBP is present in nuclei as previously reported, interaction of CTBP with certain proteins, including transcription factors or nuclear T(3) receptor, may contribute to the regulation. PMID- 11897714 TI - Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor A. AB - During bone formation and fracture healing there is a cross-talk between endothelial cells and osteoblasts. We previously showed that vascular endothelial growth factor A (VEGF-A) might be an important factor in this cross-talk, as osteoblast-like cells produce this angiogenic factor in a differentiation dependent manner. Moreover, exogenously added VEGF-A enhances osteoblast differentiation. In the present study we investigated, given the coupling between angiogenesis and bone formation, whether bone morphogenetic proteins (BMPs) stimulate osteoblastogenesis and angiogenesis through the production of VEGF-A. For this we used the murine preosteoblast-like cell line KS483, which forms mineralized nodules in vitro, and an angiogenesis assay comprising 17-d-old fetal mouse bone explants that have the ability to form tube-like structures in vitro. Treatment of KS483 cells with BMP-2, -4, and -6 enhanced nodule formation, osteocalcin mRNA expression, and subsequent mineralization after 18 d of culture. This was accompanied by a dose-dependent increase in VEGF-A protein levels throughout the culture period. BMP-induced osteoblast differentiation, however, was independent of VEGF-A, as blocking VEGF-A activity by a VEGF-A antibody or a VEGF receptor 2 tyrosine kinase inhibitor did not affect BMP-induced mineralization. To investigate whether BMPs stimulate angiogenesis through VEGF A, BMPs were assayed for their angiogenic activity. Treatment of bone explants with BMPs enhanced angiogenesis. This was inhibited by soluble BMP receptor 1A or noggin. In the presence of a VEGF-A antibody, both unstimulated and BMP stimulated angiogenesis were arrested. Conditioned media of KS483 cells treated with BMPs also induced a strong angiogenic response, which was blocked by antimouse VEGF-A but not by noggin. These effects were specific for BMPs, as TGF beta inhibited osteoblast differentiation and angiogenesis while stimulating VEGF A production. These findings indicate that BMPs stimulate angiogenesis through the production of VEGF-A by osteoblasts. In conclusion, VEGF-A produced by osteoblasts in response to BMPs is not involved in osteoblast differentiation, but couples angiogenesis to bone formation. PMID- 11897716 TI - Identification and characterization of a functionally distinct form of human estrogen receptor beta. AB - Estrogen receptors are important for the development and maintenance of many different tissues in the body including the breast, uterus, brain and bone. There are two known genes encoding estrogen receptors, Estrogen Receptor alpha (ER alpha) and Estrogen Receptor beta (ER beta). These receptors are transcription factors with distinct functional domains involved in DNA binding, ligand binding and transcriptional regulation. A novel isoform of human ER beta (ER beta 548) which includes an extended amino terminal domain has been identified. Isoform specific antibodies confirm the presence of this receptor in human tissue. Transactivation analysis with different estrogenic ligands indicates that ER beta 548 is functionally distinct from previously reported forms of ER beta. PMID- 11897715 TI - Thyroid function in mice with compound heterozygous and homozygous disruptions of SRC-1 and TIF-2 coactivators: evidence for haploinsufficiency. AB - Steroid receptor coactivator (SRC)-1 and transcriptional intermediary factor (TIF)-2 are homologous nuclear receptor coactivators. We have investigated their possible redundancy as thyroid hormone (TH) coactivators by measuring thyroid function in compound SRC-1 and TIF-2 knock out (KO) mice. Whereas SRC-1 KO (SRC 1(-/-)) mice are resistant to TH and SRC-1(+/-) are not, we now demonstrate that TIF-2 KO (TIF-2(-/-)) mice have normal thyroid function. Yet double heterozygous, SRC-1(+/-)/TIF-2(+/-) mice manifested resistance to TH of a similar degree as that in mice completely deficient in SRC-1. KO of both SRC-1 and TIF-2 resulted in marked increases of serum TH and thyrotropin concentrations. This work demonstrates gene dosage effect in nuclear coactivators manifesting as haploinsufficiency and functional redundancy of SRC-1 and TIF-2. PMID- 11897717 TI - Endocrine-related resources from the National Institutes of Health. PMID- 11897718 TI - National Hormone & Peptide Program--National Institute of Diabetes and Digestive and Kidney Diseases: recombinant hormones, hypothalamic peptides & other hormones, antisera, reagents, & hormone assay services available. PMID- 11897721 TI - Should physicians support the medical use of marijuana? Yes: it can be effective when all else fails. Point. PMID- 11897722 TI - Should physicians support the medical use of marijuana? No: evidence of its safety and efficacy is weak. Counterpoint. PMID- 11897723 TI - Cloning human embryos: decisions must not be made by private corporations behind closed doors. PMID- 11897724 TI - Surf's up! Protecting the privacy of health information on the Internet: we need new privacy laws and better encryption of information. PMID- 11897725 TI - Are we there yet? The road to a malaria vaccine. PMID- 11897726 TI - Breast cancer characteristics of Vietnamese women in the Greater San Francisco Bay Area. AB - OBJECTIVES: To examine breast cancer characteristics of women of Vietnamese ancestry living in the San Francisco Bay Area in comparison with those of other racial or ethnic groups in the same area. DESIGN: Data were obtained from the population-based Greater Bay Area Cancer Registry, part of the Surveillance, Epidemiology, and End Results program. We included breast cancer cases diagnosed from 1988 to 1999 and compared the age at diagnosis, stage and histologic grade at diagnosis, estrogen- and progesterone-receptor status, and surgery types across racial or ethnic groups. We also modeled the effect of patient and clinical characteristics and hospital and physician on the racial or ethnic variations in surgery type. RESULTS: Vietnamese women were younger at diagnosis than other racial or ethnic subgroups (mean age, 51.0 years), with 49.6% of the diagnoses occurring in patients younger than 50. They were also significantly more likely to have received mastectomy for their in situ and localized tumors (61.1% having mastectomy) than women of other racial or ethnic groups. The increased likelihood of having mastectomy among Vietnamese women was not affected greatly by age, year of diagnosis, tumor stage, histologic grade, or physician, but was partly attributable to the hospital of diagnosis. CONCLUSIONS: The effects of a lower mean age at diagnosis and the reasons for an unexpectedly higher percentage of mastectomies in this Asian subgroup should be further explored. PMID- 11897727 TI - Breast cancer in Asian women. PMID- 11897729 TI - Patient education for H pylori: who benefits? PMID- 11897728 TI - Helicobacter pylori eradication in dyspeptic primary care patients: a randomized controlled trial of a pharmacy intervention. AB - OBJECTIVE: To determine the effectiveness of structured adherence counseling by pharmacists on the eradication of Helicobacter pylori when using a standard drug treatment regimen. DESIGN: Randomized controlled clinical trial. SETTING: Nonprofit group-practice health maintenance organization (HMO). PARTICIPANTS: HMO primary care providers referred 1,393 adult dyspeptic patients for carbon 14 urea breath testing (UBT). INTERVENTIONS: Those whose tests were positive for H pylori (23.3%) were provided a standard antibiotic regimen and randomly assigned to receive either usual-care counseling from a pharmacist or a longer adherence counseling session and a follow-up phone call from the pharmacist during drug treatment. All subjects were given the same 7-day course of omeprazole, bismuth subsalicylate, metronidazole, and tetracycline hydrochloride (OBMT). Dyspepsia symptoms were recorded at baseline and following therapy. OUTCOMES: The main outcome was eradication of H pylori as measured by UBT at 3-month follow-up. Secondary outcomes were patient satisfaction and dyspepsia symptoms at 3-month follow-up. RESULTS: Of the 333 participants randomly assigned to treatment, 90.7% completed the 3-month follow-up UBT and questionnaires. Overall eradication rate with the OBMT regimen was 80.5% with no significant difference in eradication rates between the 2 groups (P=0.98). Conclusions In this study, additional counseling by pharmacists did not affect self-reported adherence to the treatment regimen, eradication rates, or dyspepsia symptoms but did increase patient satisfaction. PMID- 11897731 TI - Abdominal pain in children and the diagnosis of appendicitis. PMID- 11897730 TI - Dyspepsia, peptic ulcer disease, and esophageal reflux disease. PMID- 11897732 TI - Palliative surgery for cancer pain. PMID- 11897733 TI - Should all patients with diabetes mellitus be screened for hemochromatosis? PMID- 11897734 TI - Diabetes and HFE mutations: cause or coincidence? PMID- 11897735 TI - Equipping primary care physicians for the digital age. The Internet, online education, handheld computers, and telemedicine. PMID- 11897736 TI - Death--whose decision? Physician-assisted dying and the terminally ill. PMID- 11897737 TI - The Robert Wendland case. PMID- 11897738 TI - Legal implications of the Wendland case for end-of-life decision making. PMID- 11897739 TI - The ethics of surrogate decision making. PMID- 11897740 TI - What is a minimally conscious state? PMID- 11897742 TI - "The price of freedom is visible here". PMID- 11897741 TI - Medications for smoking cessation. PMID- 11897743 TI - Are medical students ready for the digital age? PMID- 11897744 TI - Trauma ICU: sutures, scalpels, and swears. PMID- 11897745 TI - Shaping America's health care professions: how the health sector will respond to "generation X". PMID- 11897746 TI - Why the disease-based model of medicine fails our patients. PMID- 11897747 TI - Is lithium still worth using? An update of selected recent research. AB - The treatment of bipolar disorder has seen greater innovation in the past decade than at any other time since the introduction of lithium and the neuroleptics a half-century ago. The place of lithium in contemporary psychiatric therapeutics has become controversial, calling for the present overview of research findings pertaining to its use in treating patients with bipolar disorder. Lithium, by itself, typically is inadequate for rapid control of acute mania; antipsychotics, divalproex, or potent sedatives are commonly used, with or without lithium, for this purpose. The special usefulness of lithium lies in long-term prevention of recurrences of mania and bipolar depression and in reducing risk of suicidal behavior. Lithium also may be beneficial in recurrent unipolar depression and is an effective adjunct for treatment-resistant depression. Expectations that prolonged untreated bipolar illness, multiple episodes, rapid cycling, or retreatment following discontinuation might routinely lead to lithium nonresponsiveness, and the belief that lithium is too toxic for use during pregnancy, have not been borne out by research. Lithium retains a substantial share of prescriptions for bipolar disorder and is inexpensive. No other treatment has performed as well as lithium in as many aspects of long-term care of bipolar disorder patients, and despite some risks and limitations, lithium remains the standard against which all proposed alternatives are compared. PMID- 11897748 TI - Women and alcohol use disorders. AB - Although substance abuse and dependence have been increasing among women in the United States for some time, only during the past two decades have researchers started to focus on women and alcohol use disorders. In the past all-male samples were generally used because they were much more easily available; when mixed gender populations were examined, women were often underrepresented. Gender bias was evident in research on alcohol dependence even in the early 1990s. A critical review of addiction specialty journals in 1995 concluded that researchers still commonly used male populations and generalized the findings to both sexes. Recent studies on gender differences in alcohol use disorders have found that compared to men, women become intoxicated after drinking half as much, metabolize alcohol differently, develop cirrhosis of the liver more rapidly, and have a greater risk of dying from alcohol-related accidents. This article reviews the existing literature, focusing on four central questions: (1) Are alcohol use disorders becoming increasingly prevalent in women, thereby closing the gender gap between men and women? (2) Do the physical effects of alcohol differ by gender, and if so, why? (3) Do men and women differ in frequency and type of treatment services sought for alcohol use disorders? (4) What role does gender play in the process of recovery from alcohol dependence? PMID- 11897749 TI - The neural network basis for abnormalities of attention and executive function in major depressive disorder: implications for application of the medical disease model to psychiatric disorders. AB - The deficits in attention and executive function characteristic of major depressive disorder (MDD) are reviewed. The networks underlying attention and executive function, the neuropsychological tests commonly used to evaluate these domains, and the neuroanatomy of MDD are also discussed. A neural network approach to the attentional and executive function deficits of MDD has ramifications for hypothesis-guided research, the cognitive model of depression, and application of the medical disease model to psychiatric disorders. PMID- 11897750 TI - Grief, psychosis, and panic intervention with a psychotic patient: integrating psychodynamic and cognitive-behavioral approaches. PMID- 11897751 TI - Strategies for treatment of SSRI-associated sexual dysfunction: a survey of an academic psychopharmacology practice. PMID- 11897752 TI - Psychotherapeutic considerations in treating Latinos. PMID- 11897753 TI - Psychiatric therapeutics and "the public" in England in the eighteenth and nineteenth centuries. PMID- 11897754 TI - Central pulse pressure and mortality in end-stage renal disease. AB - Damage of large arteries is a major factor in the high cardiovascular morbidity and mortality of patients with end-stage renal disease (ESRD). Increased aortic pulse wave velocity (PWV) and brachial pulse pressure (PP) are the principal arterial markers of cardiovascular mortality described in these patients. Whether central (carotid) PP and brachial-carotid PP amplification may predict all-cause (including cardiovascular) mortality has never been investigated. A cohort of 180 patients with ESRD who were undergoing hemodialysis was studied between January 1990 and March 2000. The mean duration of follow-up was 52+/-36 months (mean+/ SD). Mean age at entry was 51.5+/-16.3 years. Seventy deaths occurred, including both cardiovascular and noncardiovascular fatal events. At entry, patients underwent carotid PP measurements (pulse wave analysis), echocardiography, and aortic PWV (Doppler ultrasonography), together with standard clinical and biochemical analyses. On the basis of Cox analyses, after adjustment of age, time on dialysis before inclusion, and previous cardiovascular events, 3 factors emerged as predictors of all-cause mortality: carotid PP, brachial/carotid PP, and aortic PWV. Adjusted hazard ratios for 1-SD increments were 1.4 (1.1 to 1.8) for carotid PP, 0.5 (0.3 to 0.8) for brachial/carotid PP, and 1.3 (1.0 to 1.7) for PWV. Brachial blood pressure, including PP, had no predictive value for mortality after adjustment. These results provide the first direct evidence that in patients with ESRD, the carotid PP level and, mostly, the disappearance of PP amplification are strong independent predictors of all-cause (including cardiovascular) mortality. PMID- 11897755 TI - Correlates of left atrial size in hypertensive patients with left ventricular hypertrophy: the Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) Study. AB - Left ventricular hypertrophy has been suggested to mediate the relation between hypertension and left atrial enlargement, with associated risks of atrial fibrillation and stroke. However, less is known about correlates of left atrial size in hypertensive patients with left ventricular hypertrophy. We assessed left atrial size by echocardiography in 941 hypertensive patients, age 55 to 80 (mean, 66) years, with electrocardiographic left ventricular hypertrophy at baseline in the Losartan Intervention For Endpoint reduction in hypertension study. Enlarged left atrial diameter (women, >3.8 cm; men, >4.2 cm) was present in 56% of women and 38% of men (P<0.01). Compared with the 512 patients with normal left atrial size, the 429 patients with enlarged left atrium more often had mitral regurgitation, atrial fibrillation, and echocardiographic left ventricular hypertrophy. They also had higher age, systolic blood pressure, pulse pressure, weight, body mass index, left ventricular internal chamber dimension, stroke volume, and mass and lower relative wall thickness and ejection fraction (all, P<0.05). In logistic regression analysis, left atrial enlargement was related to left ventricular hypertrophy and eccentric geometry; greater body mass index, systolic blood pressure, and age; female gender; mitral regurgitation; and atrial fibrillation (all, P<0.05). Thus, left atrial size in hypertensive patients with electrocardiographic left ventricular hypertrophy is influenced by gender, age, obesity, systolic blood pressure, and left ventricular geometry independently of left ventricular mass and presence of mitral regurgitation or atrial fibrillation. PMID- 11897756 TI - Prevalence of left ventricular hypertrophy in hypertensive patients without and with blood pressure control: data from the PAMELA population. Pressioni Arteriose Monitorate E Loro Associazioni. AB - Previous studies have shown that in the population, only a minority of treated hypertensive patients achieve blood pressure (BP) control. Whether and to what extent this inadequate control has reflection on hypertension-related organ damage has never been systematically examined. In 2051 subjects belonging to the PAMELA (Pressioni Arteriose Monitorate E Loro Associazioni) Study population, we measured office, home, and 24-hour ambulatory BP values, together with echocardiographic left ventricular mass and wall thickness. Based on the fraction on antihypertensive treatment and on measurements of increased or normal office, home, or 24-hour ambulatory BP values, subjects were classified as normotensives, untreated hypertensives, treated hypertensives with inadequate BP control, and treated hypertensives with effective BP control. Compared with values in the normotensive group, left ventricular mass index, left ventricular wall thickness, and prevalence of left ventricular hypertrophy were markedly increased not only in untreated hypertensive patients but also in treated hypertensives with inadequate BP control. Echocardiographic abnormalities were less in treated hypertensives with BP control than in patients with inadequate BP control, but values were still clearly greater than in normotensive subjects. This was the case regardless whether BP control was assessed by office, home, and/or ambulatory values. Our data provide evidence that in the hypertensive fraction of the population, cardiac structural alterations can be frequently found in both the presence and absence of antihypertensive treatment. They also imply that even effective treatment of hypertension does not allow complete reversal of the cardiac organ damage characterizing high BP states. PMID- 11897757 TI - Assessment of left ventricular mass by cardiovascular magnetic resonance. AB - Left ventricular hypertrophy is associated with significant excess mortality and morbidity. The study and treatment of this condition, in particular the prognostic implications of changes in left ventricular mass, require an accurate, safe, and reproducible method of measurement. Cardiovascular magnetic resonance is a suitable tool for this purpose, and this review assesses the technique in comparison with others and examines the clinical and research implications of the improved reproducibility. PMID- 11897759 TI - Blood pressure response to heart rate during exercise test and risk of future hypertension. AB - Previous works have shown that exaggerated blood pressure response to exercise is a valid risk marker for future hypertension, yet the use of an exercise test as a means of early prediction of hypertension still requires methodological development and confirmation. The purpose of this study was to determine abnormal ranges of blood pressure responses in relation to heart rate increase during exercise and to examine the clinical utility of exercise blood pressure measurement in evaluating individual risk for developing hypertension. We examined exercise test data from a population-based sample of 1033 nonmedicated normotensive men (mean age, 42.9+/-8.5 years; range, 20 to 59 years). Percentile curves of systolic and diastolic blood pressure responses to relative heart rate increments during submaximal exercise were constructed using a third-order polynomial model with multiple regression analysis. Of the original study sample, a cohort of 726 subjects was followed for hypertensive outcome for an average period of 4.7 years. Progression to hypertension, defined as a blood pressure of > or =140/90 mm Hg or the initiation of antihypertensive therapy, was found in 114 subjects (15.4%). Kaplan-Meier survival estimates showed that the cumulative incidence of hypertension increased progressively with higher percentiles of systolic and diastolic blood pressure response (both, P<0.01). A Cox proportional survival analysis revealed a significantly increased risk for developing hypertension associated with exaggerated blood pressure response to exercise after multivariable adjustments for traditional risk factors (relative risk, 3.8; 95% confidence interval, 2.3 to 6.1). These results suggest that an exaggerated blood pressure response to heart rate during exercise is predictive of future hypertension independent of other important risk factors and lend further support to the concept that blood pressure measurement during exercise test is a valuable means of identifying normotensive individuals at high risk for developing hypertension. PMID- 11897758 TI - Aldosterone and D-glucose stimulate the proliferation of human cardiac myofibroblasts in vitro. AB - The renin-angiotensin-aldosterone-system appears to be involved in the development of cardiac fibrosis in rodents, characterized by nonepithelial cell proliferation and changes in the extracellular matrix. The aim of our study was to investigate the effect of high aldosterone concentrations on the proliferation of human cardiac interstitial cells in vitro. In addition, the effect of D glucose as another risk factor for fibrosis, eg, in the diabetic heart, was investigated. Human cardiac myofibroblast cultures were established, and growth rates were measured by WST-1 assay in fetal calf serum-free Dulbecco's modified Eagle's medium (DMEM). Cells in culture showed a significant increase in number between 24 to 72 hours of cultivation under basal conditions (DMEM, 10% fetal calf serum). Aldosterone at high concentrations (10(-8) and 10(-7) mol/L) significantly (P<0.01) increased the proliferation of cultured cardiac myofibroblasts. Comparable effects were observed after incubation of the cells with high D-glucose concentrations (15 and 25 mmol/L, P<0.01). No additive growth stimulation was evident when the cells were incubated in medium containing both aldosterone and D -glucose. These results suggest a role for aldosterone and glucose in mediating the cardiac fibrosis through stimulation of myofibroblast growth in patients with dysregulated renin-angiotensin-aldosterone-system (especially hyperaldosteronism) and impaired glucose homeostasis. PMID- 11897760 TI - Smoking impairs bradykinin-stimulated t-PA release. AB - Bradykinin stimulates tissue plasminogen activator release from human endothelium through a flow-independent, B2 receptor-dependent mechanism. The present study tests the hypothesis that smoking impairs bradykinin-stimulated tissue plasminogen activator release. Graded doses of nitroprusside (1.6 to 6.4 microg/min), methacholine (3.2 to 12.8 microg/min), and bradykinin (100 to 400 ng/min) were infused in the brachial artery in random order in 20 smokers and 12 nonsmokers matched for age, gender, and body mass index. Forearm blood flow was measured by strain-gauge plethysmography. All 3 drugs caused a dose-dependent increase in forearm blood flow, with no significant difference between smokers and nonsmokers. Bradykinin (P=0.001) and methacholine (P=0.001) caused significant dose-dependent increases in net tissue plasminogen activator release. The tissue plasminogen activator response to bradykinin was significantly greater than the tissue plasminogen activator response to methacholine in the nonsmokers (maximal net tissue plasminogen activator release, 73.2+/-21.5 versus 27.6+/-7.2 ng/min per 100 mL; P=0.001) but not in the smokers (maximal net tissue plasminogen activator release, 44.5+/-10.7 versus 24.8+/-9.3 ng/min per 100 mL; P=0.154). The effect of bradykinin (P=0.037), but not methacholine (P=0.978), on net tissue plasminogen activator release was significantly reduced in smokers compared with nonsmokers. The vascular tissue plasminogen activator response to bradykinin, but not methacholine, is impaired in smokers. Stimulated tissue plasminogen activator release may be a more sensitive measure of endothelial function than vasodilation. PMID- 11897761 TI - Socioeconomic trajectories and incident hypertension in a biracial cohort of young adults. AB - We assessed the impact of initial socioeconomic status and change in socioeconomic status across 10 years, ie, status trajectories, on the development of essential hypertension among black and white young men and women. Three thousand eight hundred twenty-seven normotensive individuals ages 18 to 30 years at study entry were followed for 10 years, with blood pressure, body mass index, and socioeconomic status characteristics measured at years 0, 2, 5, 7, and 10. Socioeconomic status trajectory measures were a new educational degree earned by year 10; difficulties paying for basics during years 2 to 10; and change in income category from year 5 to 10, defined in relation to year 0 status. Hypertension was defined as systolic blood pressure > or =140, diastolic blood pressure > or =90, or antihypertensive medication use at year 10. Reporting difficulties paying for basics at study entry (odds ratio=1.45, 95% confidence interval, 1.05 to 2.02) and continued difficulties during year 2 to 10 follow-up (odds ratio=1.62, 95% confidence interval, 1.04 to 2.53) were independently associated with incident hypertension, adjusted for race-gender group, body mass index, site, age, and initial systolic blood pressure. Decline in income from year 5 to 10 tended to be associated with hypertension, P=0.07, but a new educational degree after study onset was not. Socioeconomic trajectories are independently associated with incidence of hypertension. A dynamic index of socioeconomic status may be a useful concept in understanding the effects of socioeconomic status on the natural history of hypertension. PMID- 11897762 TI - Oxidative stress in leukocytes is a possible link between blood pressure, blood glucose, and C-reacting protein. AB - Because oxidative stress and inflammation are believed to play roles in the pathogenesis of cardiovascular diseases, oxidative stress in polymorphonuclear leukocytes (PMNs) and mononuclear cells (MNCs) has been measured. A total of 529 subjects participated this study. Intracellular oxidative stress in PMNs and MNCs was measured by gated flow cytometry using carboxyfluorescin diacetate bis acetoxymethyl ester. C-reacting protein (CRP), insulin action (homeostasis model assessment), and traditional risk factors such as age, gender, body mass index, triglycerides, LDL cholesterol, HDL cholesterol, hemoglobin A(1c), and mean blood pressure were also measured. Multiple regression analysis revealed a significant correlation between mean blood pressure and PMN oxidative stress (r=0.104, P=0.018). It also demonstrated a significant correlation between hemoglobin A(1c) and PMN oxidative stress (r=0.112, P=0.021). A significant correlation was also found between CRP and MNC oxidative stress (r=0.116, P=0.008) by multiple regression analysis. In patients with both hypertension and diabetes, both PMN and MNC oxidative stress was increased (n=21, P=0.022 and P=0.006). These results suggest that both hypertension and diabetes lead to increased oxidative stress of PMNs and MNCs, and that CRP is related to MNC oxidative stress. PMID- 11897763 TI - Alteration of plasma ghrelin levels associated with the blood pressure in pregnancy. AB - Ghrelin, an endogenous ligand for the growth hormone secretagogues, was originally isolated from rat stomach. It stimulates the release of growth hormone from primary pituitary cell cultures. We investigated the plasma concentration of ghrelin peptide in 16 nonpregnant women, 18 normal pregnant women, 20 patients with pregnancy-induced hypertension, and 10 postpartum women. The plasma concentration of ghrelin in nonpregnant women was 239.5+/-16.9 fmol/mL. The plasma concentration of ghrelin in normal pregnant women at the third trimester was 127.1+/-5.6 fmol/mL. There was negative correlation between plasma ghrelin concentration and systemic blood pressure in normal pregnant women (systolic: r= 0.564, P<0.05; diastolic: r=-0.610, P<0.01). Pregnant women with pregnancy induced hypertension (177.9+/-14.6 fmol/mL, P<0.05) also had significantly higher levels of ghrelin compared with those of normal pregnant women. In addition, there was a significant correlation between plasma ghrelin levels and systemic blood pressure (systolic: r=-0.482, P<0.05; diastolic: r=-0.466, P<0.05). These results suggest for the first time that ghrelin might have some role in cardiovascular control during normal pregnancy and in pathophysiological conditions in pregnancy, such as pregnancy-induced hypertension. PMID- 11897764 TI - Nitric oxide, anti-inflammatory drugs on renal prostaglandins and cyclooxygenase 2. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used as analgesics. They inhibit cyclooxygenases (COX), preventing the formation of prostaglandins, including prostacyclin and thromboxane. A serious side effect of COX-1 and COX-2 is renal damage. We report here that both a nonselective NSAID (aspirin, acetylsalicylic acid) and COX-2 selective NSAIDs (celecoxib and NS-398) diminished renal prostacyclin and thromboxane concentration in the renal medulla. NSAIDs failed to change COX-2 and iNOS (the inducible form of NO synthase) expression. A NO donor, B-NOD, preserved renal prostacyclin and thromboxane after administration of aspirin. PGI2 and COX-2 protein were mainly expressed in the renal medulla, whereas iNOS expression was greater in the cortex. B-NOD preserved renal prostacyclin levels after administration of NSAIDs. PMID- 11897765 TI - Nitric oxide modulation of neurally induced proximal tubular fluid reabsorption in the rat. AB - This study investigated the role of NO in mediating the renal sympathetic nerve mediated increases in proximal tubular fluid reabsorption (Jva). In inactin anesthetized Wistar rats, renal sympathetic nerve stimulation (15 V, 2 ms) at 0.75 and 1.0 Hz did not change blood pressure or glomerular filtration rate but did decrease urine flow and sodium excretion in a frequency-related fashion by 40% to 50% at 1.0 Hz (both, P<0.01). Renal nerve stimulation in control animals increased Jva by 11% at 0.75 Hz (P<0.05) and 31% at 1.0 Hz (P<0.01). Intraluminal N(omega)-nitro-L-arginine methyl ester (L-NAME) resulted in a higher basal Jva (19%, P<0.05), and renal nerve stimulation had no effect on Jva. When L-NAME plus sodium nitroprusside was present intraluminally, however, there were frequency dependent increases in Jva that were similar in pattern and magnitude to the control rats. Introduction of the relatively selective nNOS blocker 7 nitroindazole intraluminally, at 10(-6) and 10(-4) M, raised basal Jva by 18% and 24%, respectively (P<0.01), and renal nerve stimulation did not change Jva. Intraluminal aminoguanidine (10(-4) M), a relatively selective iNOS blocker, did not affect basal Jva, which remained unchanged during renal nerve stimulation. These data are consistent with NO exerting a tonic inhibitory action on the basal levels of Jva, which, in part, is caused by NO generated by the nNOS isoform. Moreover, the findings have revealed that the presence of NO is necessary to ensure that renal nerves can stimulate fluid reabsorption by the proximal tubules, requiring NO generated from both nNOS and iNOS. PMID- 11897766 TI - Factors related to the occurrence of microalbuminuria during antihypertensive treatment in essential hypertension. AB - The objective of the study was to assess the factors related to the occurrence of microalbuminuria during the follow-up of a young adult group with essential hypertension that had not been previously treated. Normo-albuminuric essential hypertensives, <50 years old, who had not been previously treated with antihypertensive drugs and who did not have diabetes mellitus were included. After the initial evaluation, patients were treated using only nonpharmacological measures (n=62), beta-blockers (n=38), ACE inhibitors (n=64), calcium channel blockers (n=8), and several classes (n=15). Measurements were taken for office blood pressure, biochemical profile, and 24-hour urinary albumin excretion at the beginning of the study and were measured yearly during an average of 2.7+/-1.2 years of follow-up. Among the 187 patients included, 22 (11,7%) developed microalbuminuria (progressors, 4.4/100 patients/y). No differences were present between progressors and those who remained normo-albuminuric (nonprogressors) in terms of age, gender, body mass index, disease duration, blood pressure values, biochemical profile, familial history of diabetes or hypertension, smoking habits, or the presence of EKG left ventricular hypertrophy. The group with the lowest progression rate was the patients treated with ACE inhibitors (n=5; 2.9/100 patients/y), followed by the diet group (n=5; 3.3/100 patients/y) and the beta-blockers group (n=5; 4.1/100 patients/y). When we excluded patients treated with calcium channel blockers or those who changed over time between different classes of treatment, no significant differences in the incidence of microalbuminuria were observed among the groups. Progressors showed higher slopes of fasting glucose (4.78+/-11.4 versus 0.50+/-6.8 mg/y, P<0.02) and uric acid (0.58+/-0.93 versus 0.05+/-1.10 mg/y, P<0.03) compared with the slopes of nonprogressors. Both the slopes for glucose and systolic blood pressure over time were associated independently with the slope of the logarithm of urinary albumin excretion when adjusted for age, gender, and treatment groups. Cox proportional hazard model for progression of microalbuminuria showed that baseline urinary albumin excretion (risk ratio [RR]=1.06; confidence interval [CI] 95%, 1.01 to 1.11), slope for systolic blood pressure (RR=1.11; CI 95%, 1.03 to 1.20), and slope for glucose (RR=1.08; CI 95%, 1.03 to 1.14) were independently associated to the development of microalbuminuria. In conclusion, in a group of young adults with essential hypertension that had not been previously treated, the main factors influencing the occurrence of microalbuminuria during antihypertensive treatment were the values of microalbuminuria at baseline and the slopes for systolic blood pressure and fasting glucose. PMID- 11897767 TI - Vasodilator action of angiotensin-(1-7) on isolated rabbit afferent arterioles. AB - Recent studies have shown that angiotensin-(1-7) (Ang-[1-7]), which is generated endogenously from both Ang I and II, is a bioactive component of the renin angiotensin system and may play an important role in the regulation of blood pressure. However, little is known about its role in regulating the reactivity of the afferent arteriole or the mechanism(s) involved. We hypothesized that Ang-(1 7), acting on specific receptors, participates in the control of afferent arteriole tone. We first examined the direct effect of Ang-(1-7) on rabbit afferent arterioles microperfused in vitro, and we tested whether endothelium derived relaxing factor/NO and cyclooxygenase products are involved in its actions. To assess the vasodilator effect of Ang-(1-7), afferent arterioles were preconstricted with norepinephrine, and increasing concentrations of Ang-(1-7) were added to the lumen. We found that 10(-10) to 10(-6) mol/L Ang-(1-7) produced dose-dependent vasodilatation, increasing luminal diameter from 8.9+/-1.0 to 16.3+/-1.1 microm (P<0.006). Indomethacin had no effect on Ang-(1-7)-induced dilatation. N(G)-nitro-L-arginine methyl ester, a NO synthesis inhibitor, abolished the dilatation induced by Ang-(1-7). We attempted to determine which angiotensin receptor subtype is involved in this process. We found that 10(-6) mol/L [d-Ala7]-Ang-(1-7), a potent and selective Ang-(1-7) antagonist, abolished the dilatation induced by Ang-(1-7). An angiotensin II type 1 receptor antagonist (L158809) and an angiotensin II type 2 receptor antagonist (PD 123319) at 10(-6) mol/L had no effect on Ang-(1-7)-induced dilatation. Our results show that Ang-(1 7) causes afferent arteriole dilatation. This effect may be due to production of NO, but not the action of cyclooxygenase products. Ang-(1-7) has a receptor mediated vasodilator effect on the rabbit afferent arteriole. This effect may be mediated by Ang-(1-7) receptors, because angiotensin type 1 and type 2 receptor antagonists could not block Ang-(1-7)-induced dilatation. Thus, our data suggest that Ang-(1-7)opposes the action of Ang II and plays an important role in the regulation of renal hemodynamics. PMID- 11897768 TI - Posttranscriptional control of BNP gene expression in angiotensin II-induced hypertension. AB - B-type natriuretic peptide (BNP) plasma concentrations are raised in patients with heart failure. In several experimental models of cardiac overload, however, BNP mRNA and plasma BNP peptide levels are normal, despite the persistent increase in blood pressure and ventricular hypertrophy. In this study, the role of transcriptional mechanisms in the regulation of BNP gene expression were studied in angiotensin (Ang) II-induced hypertension by injecting DNA constructs containing the BNP promoter (-2200 to 75 bp of the transcriptional start site) linked to luciferase reporter into rat myocardium. Ang II was administered to conscious rats via intravenous infusion for 2 hours or by subcutaneous minipumps for 6 hours, 12 hours, 3 days, 1 week, and 2 weeks. Ang II increased blood pressure and cardiac mass and induced changes in diastolic function. The left ventricular BNP mRNA levels increased 2.2-fold (P<0.001) at 2 hours and peaked at 12 hours (5.2-fold, P<0.001). Thereafter, BNP mRNA levels decreased (1.8-fold induction at 3 days, P<0.05) and returned to control levels at 1 week, despite persistent hypertension and myocardial hypertrophy. Left ventricular BNP peptide concentrations followed the changes in BNP mRNA levels. The BNP promoter was activated 2.7-fold (P<0.05) at 2 hours and remained upregulated up to 2 weeks (2.8-fold, P<0.05) during Ang II infusion, except at 12 hours. These results indicate that posttranscriptional control plays a major role in the regulation of ventricular BNP gene expression in Ang II-induced hypertension. PMID- 11897769 TI - Increased methylglyoxal and oxidative stress in hypertensive rat vascular smooth muscle cells. AB - Methylglyoxal can yield advanced glycation end products via nonenzymatic glycation of proteins. Whether methylglyoxal contributes to the pathogenesis of hypertension has not been clear. The aim of the present study was to investigate whether the levels of methylglyoxal and methylglyoxal-induced advanced glycation end products were enhanced and whether methylglyoxal increased oxidative stress, activated nuclear factor-kappaB (NF-kappaB), and increased intracellular adhesion molecule-1 (ICAM-1) content in vascular smooth muscle cells from spontaneously hypertensive rats. Basal cellular levels of methylglyoxal and advanced glycation end products were more than 2-fold higher (P<0.05) in cells from hypertensive rats than from normotensive Wistar-Kyoto rats. This correlated with levels of oxidative stress and oxidized glutathione that were significantly higher in cells from hypertensive rats, whereas levels of glutathione and activities of glutathione reductase and glutathione peroxidase were significantly lower. Basal levels of nuclearly localized NF-kappaB p65 and ICAM-1 protein expression were higher in cells from hypertensive rats than from normotensive rats. Addition of exogenous methylglyoxal to the cultures induced a greater increase in oxidative stress and advanced glycation end products in cells from hypertensive rats compared with normotensive rats and significantly decreased the activities of glutathione reductase and glutathione peroxidase in cells of both rat strains. Methylglyoxal activated NF-kappaB p65 and increased ICAM-1 expression in hypertensive cells, which was inhibited by N-acetylcysteine. Our study demonstrates an elevated methylglyoxal level and advanced glycation end products in cells from hypertensive rats, and methylglyoxal increases oxidative stress, activates NF-kappaB, and enhances ICAM-1 expression. Our findings suggest that that elevated methylglyoxal and associated oxidative stress possibly contribute to the pathogenesis of hypertension. PMID- 11897770 TI - Exaggerated hypotensive effect of vascular endothelial growth factor in spontaneously hypertensive rats. AB - Vascular endothelial growth factor (VEGF) induces hypotension in normotensive subjects, which is considered to be a major side effect for treatment of ischemic diseases. However, the hypotensive effect of VEGF has not been investigated in the setting of hypertension. This study determined effects of VEGF on hemodynamics, pharmacokinetics, and release of NO and prostaglandin I2 (PGI2) in vivo and on vasorelaxation of mesentery artery rings in vitro in spontaneously hypertensive rats (SHR) compared with Wistar-Kyoto rats (WKY). Intravenous infusion of VEGF for 2 hours produced a dose-related decrease in arterial pressure, which was enhanced in conscious SHR compared with WKY (P<0.01), and an increase in heart rate in WKY but not in SHR. In response to similar doses of VEGF, compared with WKY, SHR had a higher plasma VEGF level and lower VEGF clearance (P<0.01). Circulating NO and PGI2 levels after VEGF administration were not increased in SHR versus WKY, and VEGF-induced vasorelaxation was blunted in SHR versus WKY in vitro, suggesting endothelial dysfunction in SHR. One-week VEGF infusion also caused greater hypotension (P<0.05) in the absence of tachycardia in SHR compared with WKY controls. Thus, despite blunted vasorelaxation in vitro because of endothelial dysfunction, SHR exhibited exaggerated hypotension without tachycardia in response to VEGF, which was independent of NO and PGI2. The exaggerated hypotensive response to VEGF in SHR may be owing to impaired baroreflex function and reduced VEGF clearance. The data may also suggest that more caution should be taken when VEGF is administered in patients with hypertension. PMID- 11897771 TI - Effect of endothelin blockade on basal and stimulated forearm blood flow in patients with essential hypertension. AB - Endothelin receptor antagonism lowers blood pressure in patients with essential hypertension, but the contribution of the endothelin system to increased vascular tone in these patients remains controversial. We used strain-gauge venous plethysmography to measure changes in basal and metabolically stimulated (peak reactive hyperemia [PRH]) forearm blood flow (FBF) induced by continuous intrabrachial infusion of SB 209670, a dual endothelin type A/endothelin type B receptor antagonist (ETRA) in 11 patients with hypertension and 12 healthy age matched control subjects. In both groups, ETRA caused significant vasodilation (increase in FBF compared with baseline over the last 30 minutes of ETRA infusion: 75+/-12% in control subjects, 40+/-13% in hypertension patients; P<0.01 for both groups). By repeated-measures ANOVA, there was no greater increase in FBF after ETRA in hypertension patients compared with control subjects. In addition, FBF responses to PRH, expressed as percent change from baseline (prePRH), were similar in both groups (control subjects: preETRA, 1381+/-222%; during ETRA, 921+/-178%; and hypertension patients: preETRA, 1232+/-221%: during ETRA, 865+/-285%). We conclude that basal and stimulated vasodilation induced by short-term ETRA infusion in the forearm vasculature of hypertension patients is not increased compared with that of control subjects. An enhanced contribution of the endothelin system to vascular tone in hypertension patients could not be demonstrated using this experimental approach. PMID- 11897772 TI - Serotonin-induced contraction in mesenteric resistance arteries: signaling and changes in deoxycorticosterone acetate-salt hypertension. AB - Large arteries from hypertensive subjects are hyperresponsiveness to 5 hydroxytryptamine (5-HT). We tested the hypothesis that small arteries (225 micro ID) have a profile similar to conduit arteries, including signal transduction mechanisms and the 5-HT receptor subtype(s) mediating arterial contraction in normal and high blood pressure. Aorta and mesenteric arteries from Sprague-Dawley (232+/-6 micro ID), sham (229+/-7 micro ID; systolic blood pressure, 120+/-2 mm Hg), or deoxycorticosterone acetate (DOCA)-salt rats (255+/-11 micro ID, 192+/-8 mm Hg) were mounted in a wire-based myograph. In resistance arteries from Sprague Dawley rats, the 5-HT2A receptor mediated contraction; agonists of the 5-HT1B, 5 HT1D, 5-HT1F, and 5-HT2B receptor were inactive. The tyrosine kinase inhibitor genistein (5 micromol/L, 4.8-fold rightward shift), PD 098,059 (10 micromol/L, 3.2-fold shift), phospholipase C inhibitor NCDC (100 micromol/L), and nifedipine (50 nmol/L) reduced maximum 5-HT-induced contraction in small arteries (4.5% and 53% control, respectively). As in aorta, 5-HT had a decrease in threshold (100 fold lower), increase in potency (11.6-fold leftward shift), and increase in efficacy (140% sham response) in small arteries from DOCA-salt rats compared with sham. Unlike in aorta, 5-HT-induced contraction in DOCA-salt small arteries was shifted competitively by the 5-HT2A receptor antagonist ketanserin (-log K(B) [mol/L] for both sham and DOCA-salt, 9.25+/-0.1), and contraction to the 5-HT2B agonist BW723C86 was not observed. Thus, the 5-HT2A receptor remains the contractile receptor in hypertension in small arteries. Although similarities were observed for large and small arteries, differences under the condition of DOCA-salt hypertension exist that may determine serotonergic compounds effective in lowering blood pressure. PMID- 11897775 TI - Techniques for studying arterial elastic properties. PMID- 11897773 TI - Endothelial nitric oxide synthase lies downstream from angiotensin II-induced angiogenesis in ischemic hindlimb. AB - We assessed the role of angiotensin (Ang) II in ischemia-induced angiogenesis and analyzed the molecular pathways involved in such an effect. Ischemia was produced by unilateral artery femoral occlusion in control, in valsartan-treated (Ang II receptor type I antagonist, 20 mg/kg per day), in Ang II-treated (5 ng/kg per min), and in Ang II and valsartan-treated rats. After 28 days, angiogenesis was assessed by microangiography and capillary density measurement in hindlimbs. The ischemic/nonischemic leg ratio for angiographic score and capillary number increased by 2.6- and 2-fold, respectively, in Ang II-treated rats compared with controls (P<0.01). This was associated with an increase in vascular endothelial growth factor (VEGF; 1.6-fold) and endothelial NO synthase (eNOS; 1.8-fold) protein content within the ischemic leg, assessed by Western blot. Angiotensin type 1 receptor blockade and administration of VEGF neutralizing antibody (2.5 microg IP, twice a week) in Ang II-treated rats completely prevented such Ang II angiogenic effects. The key role of eNOS was then emphasized by using mice deficient in gene encoding for eNOS. In wild-type mice, Ang II (0.3 mg/kg per min) treatment increased by 1.7- and 1.6-fold the ischemic/nonischemic leg for angiographic score and blood perfusion (assessed by laser Doppler perfusion imaging) ratios, respectively (P<0.01). Conversely, no significant changes were observed in Ang II-treated mice deficient in gene encoding for eNOS. Subhypertensive dose of Ang II enhanced angiogenesis associated with tissue ischemia through angiotensin type 1 receptor activation that involved the VEGF/eNOS-dependent pathway. PMID- 11897776 TI - Electronic sphygmomanometers: are they a source of mercury in hospitals? PMID- 11897777 TI - Does acute catastrophic psychological stress disrupt diurnal cardiovascular variability? PMID- 11897778 TI - Active site mutation in DNA polymerase gamma associated with progressive external ophthalmoplegia causes error-prone DNA synthesis. AB - Progressive external ophthalmoplegia (PEO) is a heritable mitochondrial disorder characterized by the accumulation of multiple point mutations and large deletions in mtDNA. Autosomal dominant PEO was recently shown to co-segregate with a heterozygous Y955C mutation in the human gene encoding the sole mitochondrial DNA polymerase, DNA polymerase gamma (pol gamma). Since Tyr-955 is a highly conserved residue critical for nucleotide recognition among family A DNA polymerases, we analyzed the effects of the Y955C mutation on the kinetics and fidelity of DNA synthesis by the purified human mutant polymerase in complex with its accessory subunit. The Y955C enzyme retains a wild-type catalytic rate (k(cat)) but suffers a 45-fold decrease in apparent binding affinity for the incoming nucleoside triphosphate (K(m)). The Y955C derivative is 2-fold less accurate for base pair substitutions than wild-type pol gamma despite the action of intrinsic exonucleolytic proofreading. The full mutator effect of the Y955C substitution was revealed by genetic inactivation of the exonuclease, and error rates for certain mismatches were elevated by 10-100-fold. The error-prone DNA synthesis observed for the Y955C pol gamma is consistent with the accumulation of mtDNA mutations in patients with PEO. PMID- 11897779 TI - Parathyroid hormone and parathyroid hormone-related protein exert both pro- and anti-apoptotic effects in mesenchymal cells. AB - During bone formation, multipotential mesenchymal cells proliferate and differentiate into osteoblasts, and subsequently many die because of apoptosis. Evidence suggests that the receptor for parathyroid hormone (PTH) and parathyroid hormone-related protein (PTHrP), the PTH-1 receptor (PTH-1R), plays an important role in this process. Multipotential mesenchymal cells (C3H10T1/2) transfected with normal or mutant PTH-1Rs and MC3T3-E1 osteoblastic cells were used to explore the roles of PTH, PTHrP, and the PTH-1R in cell viability relative to osteoblastic differentiation. Overexpression of wild-type PTH-1R increased cell numbers and promoted osteocalcin gene expression versus inactivated mutant receptors. Furthermore, the effects of PTH and PTHrP on apoptosis were dramatically dependent on cell status. In preconfluent C3H10T1/2 and MC3T3-E1 cells, PTH and PTHrP protected against dexamethasone-induced reduction in cell viability, which was dependent on cAMP activation. Conversely, PTH and PTHrP resulted in reduced cell viability in postconfluent cells, which was also dependent on cAMP activation. Further, the proapoptotic-like effects were associated with an inhibition of Akt phosphorylation. These data suggest that parathyroid hormones accelerate turnover of osteoblasts by promoting cell viability early and promoting cell departure from the differentiation program later in their developmental scheme. Both of these actions occur at least in part via the protein kinase A pathway. PMID- 11897780 TI - Hyposmotic stress induces cell growth arrest via proteasome activation and cyclin/cyclin-dependent kinase degradation. AB - Ordered cell cycle progression requires the expression and activation of several cyclins and cyclin-dependent kinases (Cdks). Hyperosmotic stress causes growth arrest possibly via proteasome-mediated degradation of cyclin D1. We studied the effect of hyposmotic conditions on three colonic (Caco2, HRT18, HT29) and two pancreatic (AsPC-1 and PaCa-2) cell lines. Hyposmosis caused reversible cell growth arrest of the five cell lines in a cell cycle-independent fashion, although some cell lines accumulated at the G(1)/S interface. Growth arrest was followed by apoptosis or by formation of multinucleated giant cells, which is consistent with cell cycle catastrophe. Hyposmosis dramatically decreased Cdc2, Cdk2, Cdk4, cyclin B1, and cyclin D3 expression in a time-dependent fashion, in association with an overall decrease in cellular protein synthesis. However, some protein levels remained unaltered, including cyclin E and keratin 8. Selective proteasome inhibition prevented Cdk and cyclin degradation and reversed hyposmotic stress-induced growth arrest, whereas calpain and lysosome enzyme inhibitors had no measurable effect on cell cycle protein degradation. Therefore, hyposmotic stress inhibits cell growth and, depending on the cell type, causes cell cycle catastrophe with or without apoptosis. The growth arrest is due to decreased protein synthesis and proteasome activation, with subsequent degradation of several cyclins and Cdks. PMID- 11897781 TI - Contribution of human mlh1 and pms2 ATPase activities to DNA mismatch repair. AB - MutLalpha, a heterodimer composed of Mlh1 and Pms2, is the major MutL activity in mammalian DNA mismatch repair. Highly conserved motifs in the N termini of both subunits predict that the protein is an ATPase. To study the significance of these motifs to mismatch repair, we have expressed in insect cells wild type human MutLalpha and forms altered in conserved glutamic acid residues, predicted to catalyze ATP hydrolysis of Mlh1, Pms2, or both. Using an in vitro assay, we showed that MutLalpha proteins altered in either glutamic acid residue were each partially defective in mismatch repair, whereas the double mutant showed no detectable mismatch repair. Neither strand specificity nor directionality of repair was affected in the single mutant proteins. Limited proteolysis studies of MutLalpha demonstrated that both Mlh1 and Pms2 N-terminal domains undergo ATP induced conformational changes, but the extent of the conformational change for Mlh1 was more apparent than for Pms2. Furthermore, Mlh1 was protected at lower ATP concentrations than Pms2, suggesting Mlh1 binds ATP with higher affinity. These findings imply that ATP hydrolysis is required for MutLalpha activity in mismatch repair and that this activity is associated with differential conformational changes in Mlh1 and Pms2. PMID- 11897782 TI - Formation of the approximately 350-kDa Apg12-Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast. AB - Autophagy, responsible for the delivery of cytoplasmic components to the lysosome/vacuole for degradation, is the major degradative pathway in eukaryotic cells. This process requires a ubiquitin-like protein conjugation system, in which Apg12 is covalently bound to Apg5. In the yeast Saccharomyces cerevisiae, the Apg12-Apg5 conjugate further interacts with a small coiled-coil protein, Apg16. The Apg12-Apg5 and Apg16 are localized in the cytosol and pre autophagosomal structures and play an essential role in autophagosome formation. Here we show that the Apg12-Apg5 conjugate and Apg16 form a approximately 350-kDa complex in the cytosol. Because Apg16 was suggested to form a homo-oligomer, we generated an in vivo system that allowed us to control the oligomerization state of Apg16. With this system, we demonstrated that formation of the approximately 350-kDa complex and autophagic activity depended on the oligomerization state of Apg16. These results suggest that the Apg12-Apg5 conjugate and Apg16 form a multimeric complex mediated by the Apg16 homo-oligomer, and formation of the approximately 350-kDa complex is required for autophagy in yeast. PMID- 11897784 TI - The protein kinase/endoribonuclease IRE1alpha that signals the unfolded protein response has a luminal N-terminal ligand-independent dimerization domain. AB - In response to accumulation of unfolded proteins in the endoplasmic reticulum (ER), cells activate an intracellular signal transduction pathway called the unfolded protein response (UPR). IRE and PERK are the two type-I ER transmembrane protein kinase receptors that signal the UPR. The N-terminal luminal domains (NLDs) of IRE1 and PERK sense ER stress conditions by a common mechanism and transmit the signal to regulate the cytoplasmic domains of these receptors. To provide an experimental system amenable to detailed biochemical and structural analysis to elucidate the mechanism of ER-transmembrane signaling mechanism mediated by the NLD, we overexpressed the soluble luminal domain of human IRE1alpha in COS-1 cells by transient DNA transfection. Here we report the expression, purification, and characterization of the soluble NLD. The biological function of the NLD was confirmed by its ability to associate with itself and to interact with both the membrane-bound full-length IRE1alpha receptor and the ER chaperone BiP. Functional and spectral studies suggested that the highly conserved N-linked glycosylation site is not required for proper protein folding and self-association. Interestingly, we demonstrated that the NLD forms stable dimers linked by intermolecular disulfide bridges. Our data support that the luminal domain represents a novel ligand-independent dimerization domain. PMID- 11897783 TI - Recovery of visual functions in a mouse model of Leber congenital amaurosis. AB - The visual process is initiated by the photoisomerization of 11-cis-retinal to all-trans-retinal. For sustained vision the 11-cis-chromophore must be regenerated from all-trans-retinal. This requires RPE65, a dominant retinal pigment epithelium protein. Disruption of the RPE65 gene results in massive accumulation of all-trans-retinyl esters in the retinal pigment epithelium, lack of 11-cis-retinal and therefore rhodopsin, and ultimately blindness. We reported previously (Van Hooser, J. P., Aleman, T. S., He, Y. G., Cideciyan, A. V., Kuksa, V., Pittler, S. J., Stone, E. M., Jacobson, S. G., and Palczewski, K. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 8623-8628) that in Rpe65-/- mice, oral administration of 9-cis-retinal generated isorhodopsin, a rod photopigment, and restored light sensitivity to the electroretinogram. Here, we provide evidence that early intervention by 9-cis-retinal administration significantly attenuated retinal ester accumulation and supported rod retinal function for more than 6 months post-treatment. In single cell recordings rod light sensitivity was shown to be a function of the amount of regenerated isorhodopsin; high doses restored rod responses with normal sensitivity and kinetics. Highly attenuated residual rod function was observed in untreated Rpe65-/- mice. This rod function is likely a consequence of low efficiency production of 11-cis-retinal by photo-conversion of all-trans-retinal in the retina as demonstrated by retinoid analysis. These studies show that pharmacological intervention produces long lasting preservation of visual function in dark-reared Rpe65-/- mice and may be a useful therapeutic strategy in recovering vision in humans diagnosed with Leber congenital amaurosis caused by mutations in the RPE65 gene, an inherited group of early onset blinding and retinal degenerations. PMID- 11897785 TI - The crystal structure of prokaryotic phospholipase A2. AB - In this study, the x-ray crystal structures of the calcium-free and calcium-bound forms of phospholipase A(2) (PLA(2)), produced extracellularly by Streptomyces violaceoruber, were determined by using the multiple isomorphous replacement and molecular replacement methods, respectively. The former and latter structures were refined to an R-factor of 18.8% at a 1.4-A resolution and an R-factor of 15.0% at a 1.6-A resolution, respectively. The overall structure of the prokaryotic PLA(2) exhibits a novel folding topology that demonstrates that it is completely distinct from those of eukaryotic PLA(2)s, which have been already determined by x-ray and NMR analyses. Furthermore, the coordination geometry of the calcium(II) ion apparently deviated from that of eukaryotic PLA(2)s. Regardless of the evolutionary divergence, the catalytic mechanism including the calcium(II) ion on secreted PLA(2) seems to be conserved between prokaryotic and eukaryotic cells. Demonstrating that the overall structure determined by x-ray analysis is almost the same as that determined by NMR analysis is useful to discuss the catalytic mechanism at the molecular level of the bacterial PLA(2). PMID- 11897786 TI - A novel prokaryotic phospholipase A2. Characterization, gene cloning, and solution structure. AB - Until now, phospholipase A(2) (PLA(2); EC 3.1.14) has been found only from eukaryotic sources. In the present study, we found a secreted PLA(2), which is produced by a soil bacterium, Streptomyces violaceoruber A-2688, demonstrating that the enzyme is the first phospholipase A(2) identified in prokaryote. After characterization of the novel PLA(2), a gene encoding the enzyme was cloned, sequenced, and overexpressed using a Streptomyces host-vector system. The amino acid sequence showed that the prokaryotic PLA(2) has only four cysteines and less homology to the eukaryotic ones, which have 12-16 cysteines. The solution structures of the prokaryotic PLA(2), bound and unbound with calcium(II) ion, were determined by using the NMR technique and structure calculation. The overall structure of the S. violaceoruber PLA(2), which is composed of only five alpha helices, is completely different from those of eukaryotic PLA(2)s, which consist of beta-sheets and alpha-helices. The structure of the calcium-binding domain is obviously distinct from that without the ion; the ligands for the calcium(II) ion are the two carboxylates of Asp(43) (monodentate) and Asp(65) (bidentate), the carbonyl oxygen of Leu(44), and three water molecules. A calcium-binding experiment showed that the calcium dissociation constant ( approximately 5 mm) for the prokaryotic PLA(2) is much larger than those of eukaryotic ones. PMID- 11897787 TI - Protection of mice from allergen-induced asthma by selenite: prevention of eosinophil infiltration by inhibition of NF-kappa B activation. AB - The potential anti-inflammatory effect of sodium selenite in a mouse model of asthma was investigated. Selenite was injected into the peritoneum of allergen (ovalbumin)-sensitized mice before allergen challenge. Ovalbumin challenge resulted in activation of the transcription factor NF-kappaB and an increase in the expression of cell adhesion molecules (intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and E-selectin, which are encoded by NF-kappaB dependent genes) in lung tissue as well as in the recruitment of eosinophils to lung airways. These effects of ovalbumin challenge were all inhibited by pretreatment of mice with selenite. Selenite administration also increased the activity of selenium-dependent glutathione peroxidase in lung tissue. Furthermore, supplementation of A549 human airway epithelial cell cultures with selenite increased glutathione peroxidase activity as well as inhibited both the generation of hydrogen peroxide and the activation of NF-kappaB induced by tumor necrosis factor alpha in these cells. Selenite also reversed in vitro the activation of NF-kappaB induced by this cytokine in intact A549 cells. These results suggest that selenite regulates the activity of NF-kappaB by increasing the activity of glutathione peroxidase, thereby removing potential activators of NF-kappaB, and possibly also by direct oxidation of critical sulfhydryl groups of this transcription factor. These effects of selenite likely underlie its anti inflammatory action in asthma. PMID- 11897788 TI - Deletion of the NH2-terminal beta-hairpin of the ribotoxin alpha-sarcin produces a nontoxic but active ribonuclease. AB - Ribotoxins are a family of highly specific fungal ribonucleases that inactivate the ribosomes by hydrolysis of a single phosphodiester bond of the 28 S rRNA. alpha-Sarcin, the best characterized member of this family, is a potent cytotoxin that promotes apoptosis of human tumor cells after internalization via endocytosis. This latter ability is related to its interaction with phospholipid bilayers. These proteins share a common structural core with nontoxic ribonucleases of the RNase T1 family. However, significant structural differences between these two groups of proteins are related to the presence of a long amino terminal beta-hairpin in ribotoxins and to the different length of their unstructured loops. The amino-terminal deletion mutant Delta(7-22) of alpha sarcin has been produced in Escherichia coli and purified to homogeneity. It retains the same conformation as the wild-type protein as ascertained by complete spectroscopic characterization based on circular dichroism, fluorescence, and NMR techniques. This mutant exhibits ribonuclease activity against naked rRNA and synthetic substrates but lacks the specific ability of the wild-type protein to degrade rRNA in intact ribosomes. The results indicate that alpha-sarcin interacts with the ribosome at two regions, i.e. the well known sarcin-ricin loop of the rRNA and a different region recognized by the beta-hairpin of the protein. In addition, this latter protein portion is involved in interaction with cell membranes. The mutant displays decreased interaction with lipid vesicles and shows behavior compatible with the absence of one vesicle-interacting region. In agreement with this conclusion, the deletion mutant exhibits a very low cytotoxicity on human rhabdomyosarcoma cells. PMID- 11897789 TI - Phosphatidylinositol 3-kinase is required for insulin-stimulated tyrosine phosphorylation of Shc in 3T3-L1 adipocytes. AB - The interactions between the phosphatidylinositol 3-kinase (PI 3-kinase) and Ras/MAPK kinase pathways have been the subject of considerable interest. In the current studies, we find that epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) lead to rapid phosphorylation of Shc (maximum at 1-2 min), whereas insulin-mediated Shc phosphorylation is relatively delayed (maximum at 5 10 min), suggesting that an intermediary step may be necessary for insulin stimulation of Shc phosphorylation. The Src homology-2 (SH2) domain of Shc is necessary for PDGF- and EGF-mediated Shc phosphorylation, whereas the phosphotyrosine binding (PTB) domain is critical for the actions of insulin. Because the Shc PTB domain can interact with phospholipids, we postulated that PI 3-kinase might be a necessary intermediary step facilitating insulin-stimulated phosphorylation of Shc. In support of this, we found that the PI 3-kinase inhibitors, wortmannin and LY294002, blocked insulin-stimulated but not EGF- or PDGF-stimulated Shc phosphorylation. Furthermore, overexpression of a dominant negative PI 3-kinase construct (p85N-SH2) blocked insulin, but not EGF- or PDGF induced Shc phosphorylation. All three growth factors cause localization of Shc to the plasma membrane, but only the effect of insulin was inhibited by wortmannin, supporting the view that PI 3-kinase-generated phospholipids mediate insulin-stimulated Shc phosphorylation. Consistent with this, expression of a constitutively active PI 3-kinase (p110(C)(AAX)) increased membrane localization of Shc, and this was completely blocked by wortmannin. A mutant Shc with a disrupted PTB domain (Shc S154) did not localize to the membrane in p110(C)(AAX) expressing cells or after insulin stimulation and was not phosphorylated by insulin. In summary, 1) PI 3-kinase is a necessary early step in insulin stimulated Shc phosphorylation, whereas the effects of EGF and PDGF on Shc phosphorylation are independent of PI 3-kinase. 2) PI 3-kinase-stimulated generation of membrane phospholipids can localize Shc to the plasma membrane through the Shc PTB domain facilitating phosphorylation by the insulin receptor. PMID- 11897790 TI - HIV-1 integrase interaction with U3 and U5 terminal sequences in vitro defined using substrates with random sequences. AB - Successful integration of viral genome into a host chromosome depends on interaction between viral integrase and its recognition sequences. We have used a reconstituted concerted human immunodeficiency virus, type 1 (HIV-1), integration system to analyze the role of integrase (IN) recognition sequences in formation of the IN-viral DNA complex capable of concerted integration. HIV-1 integrase was presented with substrates that contained all 4 bases at 8 mismatched positions that define the inverted repeat relationship between U3 and U5 long terminal repeats (LTR) termini and at positions 17-19, which are conserved in the termini. Evidence presented indicates that positions 17-20 of the IN recognition sequences are needed for a concerted DNA integration mechanism. All 4 bases were found at each randomized position in sequenced concerted DNA integrants, although in some instances there were preferences for specific bases. These results indicate that integrase tolerates a significant amount of plasticity as to what constitutes an IN recognition sequence. By having several positions randomized, the concerted integrants were examined for statistically significant relationships between selections of bases at different positions. The results of this analysis show not only relationships between different positions within the same LTR end but also between different positions belonging to opposite DNA termini. PMID- 11897791 TI - Identification of the mitochondrial glutamate transporter. Bacterial expression, reconstitution, functional characterization, and tissue distribution of two human isoforms. AB - The mitochondrial carriers are a family of transport proteins in the inner membranes of mitochondria. They shuttle substrates, metabolites, and cofactors through this membrane and connect cytoplasm functions with others in the matrix. Glutamate is co-transported with H(+) (or exchanged for OH(-)), but no protein has ever been associated with this activity. Two human expressed sequence tags encode proteins of 323 and 315 amino acids with 63% identity that are related to the aspartate-glutamate carrier, a member of the carrier family. They have been overexpressed in Escherichia coli and reconstituted into phospholipid vesicles. Their transport properties demonstrate that the two proteins are isoforms of the glutamate/H(+) symporter described in the past in whole mitochondria. Isoform 1 is expressed at higher levels than isoform 2 in all the tissues except in brain, where the two isoforms are expressed at comparable levels. The differences in expression levels and kinetic parameters of the two isoforms suggest that isoform 2 matches the basic requirement of all tissues especially with respect to amino acid degradation, and isoform 1 becomes operative to accommodate higher demands associated with specific metabolic functions such as ureogenesis. PMID- 11897792 TI - Calcium oscillation linked to pacemaking of interstitial cells of Cajal: requirement of calcium influx and localization of TRP4 in caveolae. AB - Interstitial cells of Cajal (ICC) are considered to be pacemaker cells in gastrointestinal tracts. ICC generate electrical rhythmicity (dihydropyridine insensitive) as slow waves and drive spontaneous contraction of smooth muscles. Although cytosolic Ca(2+) has been assumed to play a key role in pacemaking, Ca(2+) movements in ICC have not yet been examined in detail. In the present study, using cultured cell clusters isolated from mouse small intestine, we demonstrated Ca(2+) oscillations in ICC. Fluo-4 was loaded to the cell cluster, the relative amount of cytosolic Ca(2+) was recorded, and ICC were identified by c-Kit immunoreactivity. We specifically detected Ca(2+) oscillation in ICC in the presence of dihydropyridine, which abolishes Ca(2+) oscillation in smooth muscles. The oscillation was coupled to the electrical activity corresponding to slow waves, and it depended on Ca(2+) influx through a non-selective cation channel, which was SK&F 96365-sensitive and store-operated. We further demonstrated the presence of transient receptor potential-like channel 4 (TRP4) in caveolae of ICC. Taken together, the results infer that the Ca(2+) oscillation in ICC is intimately linked to the pacemaker function and depends on Ca(2+) influx mediated by TRP4. PMID- 11897793 TI - Protein kinase A-independent activation of ERK and H,K-ATPase by cAMP in native kidney cells: role of Epac I. AB - This study aimed at determining the signaling pathways underlying calcitonin- and isoproterenol-induced stimulation of H,K-ATPase in rat renal collecting duct. H,K ATPase activity was determined in microdissected collecting ducts preincubated with or without either specific inhibitors or antibodies directed against intracellular signaling proteins. Transient cell membrane permeabilization with streptolysin-O allowed intracellular access of antibodies. The stimulation of H,K ATPase by calcitonin and isoproterenol was mimicked by cAMP analogues and was abolished by adenylyl cyclase inhibition. Protein kinase A inhibition abolished isoproterenol but not calcitonin effect on H,K-ATPase. Calcitonin increased the phosphorylation of extracellular signal-regulated kinase (ERK) in a protein kinase A-independent manner, and the inhibition of the ERK phosphorylation prevented the stimulation of H,K-ATPase by calcitonin. Antibodies directed against either the cAMP-activated guanine-nucleotide exchange factor Epac I, the monomeric G protein Rap-1 or the kinase Raf-B, curtailed the stimulation of H,K ATPase by calcitonin, whereas antibodies against the related monomeric G protein Ras or kinase Raf-1 had no effect. In conclusion, calcitonin stimulates H,K ATPase through a cAMP/Epac I/Rap-1/Raf-B/ERK cascade. PMID- 11897797 TI - Improved detection of apoptotic cells in archival paraffin sections: immunohistochemistry using antibodies to cleaved caspase 3. AB - Apoptosis has gained central importance in the study of many biological processes, including neoplasia, neurodegenerative diseases, and development. One of the limitations of many studies is the difficulty of specifically identifying individual apoptotic cells. Of the many specific methods developed to detect apoptotic cells, most are not applicable to histological sections of archival paraffin-embedded tissues. Recently, advances in the understanding of the molecular events in apoptosis have led to the realization that caspase activation is by far the most specific indicator of this cell suicide mechanism. Several publications have reported the development of antibodies directed at neoepitopes that are generated in various substrates through the action of caspases. One of these is that present on activated caspase 3, a ubiquitously distributed caspase that is a main effector caspase of the apoptotic cascade within cells. This study demonstrates the utility of using a recently commercially available antibody to cleaved caspase 3 in archival paraffin sections, suggesting that this may be a highly specific and sensitive method generally applicable to many studies of archival material. PMID- 11897798 TI - p73 is expressed in human thymic epithelial cells. AB - The thymus is a heterogeneous immune organ in which immature T-cells develop and eventually specialize to make certain immune responses of their own. Among various types of stromal cells in the thymus, thymic epithelial cells (TECs) have a crucially important function for presenting self-antigens and secreting cytokines to thymocytes for their maturation into T-cells. In this study we show that the p73 gene, a homologue of the tumor suppressor gene p53, was expressed in the nucleus of the human TEC in vivo and in TEC lines in vitro. Because p73 has the capacity to be a transactivator like p53, it may contribute to T-cell development in the context of TEC biology as regulated in the cell cycle and apoptosis. PMID- 11897799 TI - Comparative analysis of an improved thioflavin-s stain, Gallyas silver stain, and immunohistochemistry for neurofibrillary tangle demonstration on the same sections. AB - An improved thioflavin-S stain, Gallyas silver stain, and two immunostainings were quantitatively compared for demonstration of neurofibrillary tangles (NFTs) on the same sections. Sections of hippocampal formation from seven cases of Alzheimer's disease (AD) were immunofluorescently stained with a commercially available polyclonal NFT antibody or a PHF-1 monoclonal antibody, followed by an improved thioflavin-S stain, and finally by Gallyas silver staining. The thioflavin-S method was improved by using a combination quenching method that removes background autofluorescence without remarkable tissue damage and by post treatment with concentrated phosphate buffer, which minimizes photobleaching. PHF 1 or NFT immunostaining is much less sensitive than the improved thioflavin-S staining and Gallyas silver staining, particularly in the transentorhinal region. Moreover PHF-1 immunoreactivity varied greatly among AD individuals. Thioflavin-S staining and Gallyas silver staining show almost the same sensitivity in NFT demonstration, but only the former depends on the secondary protein structure of NFTs. This study suggests that the improved thioflavin-S staining is a simple, sensitive, and consistent method for demonstration of neurofibrillary pathology. PMID- 11897800 TI - Expression of invariant chain (CD 74) and major histocompatibility complex (MHC) class II antigens in the human fetus. AB - During the initiation of an immune response, antigen-presenting cells employ MHC class II antigens as key molecules to present small peptides to CD4-positive lymphocytes. The invariant chain (Ii; CD74) plays a critical role in this process by influencing the expression and peptide loading of the MHC class II molecules. Therefore, coordinate expression of these molecules is believed to play an important role in antigen presentation. This study explores the expression of these molecules in fetal tissues. Formalin-fixed, paraffin-embedded multi-organ tissue blocks from aborted fetuses (age range 7-22 weeks) were immunostained for Ii/CD74 and MHC class II antigens using commercially available monoclonal antibodies for Ii/CD74 (LN2) and MHC class II antigens (LN3), respectively. Coordinate staining for Ii/CD74 and MHC class II antigens was seen in the skin, proximal renal tubules, tips of small intestinal mucosa, and cells of the reticuloendothelial system, including the spleen and thymus. Expression of Ii/CD74, but not of MHC class II antigens, was seen in pulmonary alveolar epithelium in all cases and in testicular Leydig cells (11 of 11 testes examined). The distribution and intensity of staining did not change significantly with age. In conclusion, this study describes distribution of Ii/CD74 and MHC class II antigens in human fetal tissues. Coordinate expression of Ii/CD74 and MHC class II antigens was identified in most fetal tissues, but there were also notable exceptions. In all cases this took the form of expression of Ii/CD74 in the absence of MHC class II expression. Discordance was particularly striking in pulmonary alveolar epithelium and testicular Leydig cells. This suggests that the Ii/CD74 molecule has functional roles in addition to its role in antigen presentation. PMID- 11897801 TI - Stanniocalcin 1 (STC1) protein and mRNA are developmentally regulated during embryonic mouse osteogenesis: the potential of stc1 as an autocrine/paracrine factor for osteoblast development and bone formation. AB - STC1, a mammalian homologue of stanniocalcin (STC) which plays a major role in calcium/phosphate homeostasis in fish, has been recently isolated. We have characterized the spatiotemporal distribution of STC1 mRNA and protein during mouse embryonic development generally and osteogenesis specifically. Northern blotting analysis of whole embryos showed that STC1 mRNA is highly and differentially expressed during embryogenesis. By in situ hybridization, STC1 mRNA was detected early in mesenchymal condensations and was then found to be highly expressed in perichondrial cells, periosteal cells, and then osteoblasts during endochondral bone formation. In bones forming by intramembranous ossification, STC1 mRNA was not detected until osteogenic cells appeared. The cellular distribution of STC1 protein closely corresponded to that of its mRNA, but the protein was also detected in hypertrophic chondrocytes. In the MC3T3-E1 osteogenic cell model, STC1 protein and mRNA were detectable throughout proliferation and differentiation stages but levels were relatively higher late during nodule formation/mineralization phases. For comparison, STC1 mRNA was also found in epithelial cells of both embryonic and adult intestine that had not previously been described among tissues responsive to calcium/phosphate transport. These results suggest that STC1 is expressed in a time- and cell specific manner and may play an autocrine/paracrine role during osteoblast development and bone formation. PMID- 11897802 TI - Structure of outer hair cell stereocilia side and attachment links in the chinchilla cochlea. AB - The structure and symmetry of chinchilla outer hair cell (OHC) stereocilia side and attachment links were investigated by transmission electron microscopy using tannic acid and Cuprolinic blue histochemical procedures. The side links run laterally between and across the rows of the stereocilia and connect the stereocilia together within the bundle. Attachment links form a crown-like array around the tips of only the tallest OHC stereocilia and attach these stereocilia to the Type B fibrils of the tectorial membrane. Computer averaging of the side links from tannic acid-treated tissues showed a central dense region of the link between adjacent stereocilia and a smaller dense portion at the plasma membrane end of the link. Computer averaging of Cuprolinic blue-treated tissues showed low electron density of the central region of the link, and the plasma membrane ends of the link were electron dense. After tannic acid treatment, the attachment links showed a diffused radial distribution around the tips of the tallest OHC stereocilia. After Cuprolinic blue treatment, the attachment links appeared as electron-dense, membrane-bound granular structures arranged with radial symmetry. The central regions of the side links are reactive to tannic acid. These regions appear to contain neutral and basic residues of proteins and participate in side by-side association of the side links in regular aggregates. Cuprolinic blue reactive regions of the side and attachment links appear to contain acidic sulfated residues of glycoproteins or proteoglycans, which may be involved in the attachment of these links to the stereocilium membrane. PMID- 11897803 TI - Early intracellular events during internalization of Listeria monocytogenes by J774 cells. AB - The gram-positive bacillus Listeria monocytogenes gains entry into host cells through a phagosome membrane that forms around entering bacteria. During the early stages of internalization the invading bacteria appear to modify the protein composition of the forming phagosome membrane in J774 cells. MHC class II molecules on the cell surface and exposed surface molecules available for biotinylation are excluded from the bacteria-host cell membrane interface and from the forming phagosome. This exclusion of MHC class II molecules from the early phagosome may partially help to explain previous reports suggesting that L. monocytogenes is able to interfere with antigen presentation. Inside the host cell, MHC class II molecules are delivered to the phagosome membrane. This is followed by delivery of LAMP 1, a marker of late endocytic compartments, and fusion with low-pH compartments. The bacteria then escape into the cell cytoplasm, possibly assisted by rapid delivery of this low-pH environment. PMID- 11897804 TI - Distribution of melatonin MT1 receptor immunoreactivity in human retina. AB - Melatonin is synthesized in the pineal gland and retina during the night. Retinal melatonin is believed to be involved in local cellular modulation and in regulation of light-induced entrainment of circadian rhythms. The present study provides the first immunohistochemical evidence for the localization of melatonin 1a-receptor (MT1) in human retina of aged subjects. Ganglion, amacrine, and photoreceptor cells expressed MT1. In addition, MT1 immunoreactivity was localized to cell processes in the inner plexiform layer and to central vessels of the retina, as well as to retinal vessels but not to ciliary or choroidal vessels. These results support a variety of cellular and vascular effects of melatonin in the human retina. Preliminary evidence from patients with Alzheimer's disease (AD) revealed increased MT1 immunoreactivity in ganglion and amacrine cells, as well as in vessels. In AD cases photoreceptor cells were degenerated and showed low MT1 expression. PMID- 11897805 TI - Taurine in submandibular gland of the rat: effect of muscarinic drugs. AB - Taurine exerts a number of actions in mammalian cells, including regulation of ion transport and osmoregulation. The production and secretion of saliva involve transepithelial ion transport, thereby making the plasma-like primary saliva hypotonic before secretion. Therefore, it is plausible to suggest modulation of salivary taurine by muscarinic agents that affect salivary gland function. One of the objectives of this study was to determine tissue content and localization of taurine in the submandibular gland of the rat. Further, we determined whether treatment with muscarinic drugs that either increase (e.g., pilocarpine) or decrease (e.g., propantheline) saliva secretion affects the submandibular gland taurine content. The results indicate that the submandibular gland contains an appreciable amount of taurine (8.9 +/- 0.3 micromoles/g wet wt). Further, acute treatment of the rats with either of the muscarinic drugs did not significantly affect tissue taurine content compared to the control group. By contrast, chronic treatment with propantheline, but not pilocarpine, reduced the tissue content of taurine compared to the control rats (p<0.05). Utilizing light microscopic immunohistochemical techniques, intense immunoreactivity was found primarily in the striated ducts of the submandibular gland. Neither pilocarpine nor propantheline treatment led to differential distribution of immunoreactivity in this tissue. In conclusion, the submandibular gland contains an appreciable amount of taurine, primarily in the striated ducts, that can be decreased by chronic muscarinic receptor blockade. PMID- 11897806 TI - Expression and distribution of vascular endothelial growth factor receptor Flk-1 in the rat pituitary. AB - Vascular endothelial growth factor (VEGF) acts primarily as an endothelial cell mitogen via the specific receptors Flk-1 and Flt-1. To help further define the possible role of VEGF in the control of pituitary cell function, we examined Flk 1 expression in normal rat pituitaries and in GH3 cells. Flk-1 expression was studied by immunohistochemistry, in situ hybridization, and double-labeling immunofluorescence combined with confocal laser microscopy. In normal rat pituitaries, Flk-1-immunoreactive cells appeared widely distributed only in the anterior lobe and were not detected in the intermediate or posterior lobe. Apart from the adenohypophysial cells, Flk-1 immunopositivity was also evident in endothelial cells of many capillaries distributed throughout the gland. Immunohistochemistry also showed that majority of GH3 cells expressed Flk-1 protein. In situ hybridization showed conclusive staining with the antisense probe and confirmed the immunohistochemical results. The double immunofluorescence method revealed Flk-1 expression in all types of hormone producing adenohypophysial cells but not in folliculostellate cells. The percentage of immunopositive cells varied among the various cell types. The present study demonstrates that pituitary cells are not only sources of VEGF but also targets of this multifunctional substance, supporting the concept that VEGF functions as an autocrine/paracrine factor in the pituitary. PMID- 11897807 TI - Improvement of non-radioactive in situ hybridization in human airway tissues: use of PCR-generated templates for synthesis of probes and an antibody sandwich technique for detection of hybridization. AB - We describe the use of non-traditional methods of probe synthesis and quantification and detection of hybridization that appreciably improved non radioactive in situ hybridization (ISH) in human airway tissue. To avoid the problems of bacterial cloning, plasmid digestion, and probe hydrolysis, we synthesised complementary RNA probes (riboprobes) for ISH from PCR-generated DNA. DNA template was produced by nested PCR incorporation of T7 and SP6 RNA polymerase promoters. We then compared the efficiency of in vitro transcription from PCR-generated template with traditional plasmid template by quantifying the relative probe fluorescence in denaturing gels. Transcription with SP6 or T7 polymerase in either orientation produced TNF riboprobes from a single PCR generated template more efficiently than from plasmid, providing there were no primer hairpin loops. Fluorescence quantification enabled equal amounts of probe label to be used in ISH, eliminating signals from the sense probe and demonstrating that probes transcribed from PCR templates were as sensitive as hydrolyzed probe transcribed from plasmid. Detection of ISH by a conventional anti-hapten, alkaline phosphatase-based technique was found to cause tissue damage due to extended substrate incubation at high pH. We therefore developed a four-layer, avidin-biotin-peroxidase technique that afforded greater sensitivity, allowing brief substrate incubation and resulting in structural preservation of tissue. PMID- 11897808 TI - Localization of NTPDase1/CD39 in normal and transformed human pancreas. AB - Elevated levels of extracellular ATP have been observed in many tumors. We have localized NTPDase1/CD39, one of the principal extracellular nucleotide hydrolyzing enzymes, in normal and cancerous human pancreas. NTPDase/E-ATPDase activity was demonstrated with an enzyme histochemical technique on cryosections of human pancreas. Acinar and duct epithelial cells were devoid of E-ATPDase activity in both normal and transformed tissue. Endothelial cells and smooth muscle around blood vessels and larger ducts showed strong activity. Nerves, connective tissue, and the beta-cells of the islets were also stained. In cancerous tissue this activity was diminished in the smooth muscle around the ducts and was absent from newly formed connective tissue. Immunostaining for CD39 supported these results but revealed the presence of inactive CD39 in the duct epithelial cells. We hypothesize that the significantly diminished activity of NTPDase1 in the tissues surrounding the ducts may be associated with the processes that lead to tumor formation in human pancreas. PMID- 11897809 TI - ERp29 is a ubiquitous resident of the endoplasmic reticulum with a distinct role in secretory protein production. AB - ERp29 was recently characterized biochemically as a novel protein that resides in mammalian endoplasmic reticulum (ER). Here we applied immunochemical procedures at the cellular level to investigate the hypothesized role of ERp29 in secretory protein production. ERp29 was localized exclusively to the ER/nuclear envelope of MDCK cells using confocal immunocytochemistry and comparative markers of the ER lumen, ER/Golgi membrane, nuclei, and mitochondria. A predominant association with rough ER was revealed by sucrose-gradient analysis of rat liver microsomes. Immunohistochemistry showed ERp29 expression in 35 functionally distinct cell types of rat, establishing ERp29 as a general ER marker. The ERp29 expression profile largely paralleled that of protein disulfide isomerase (PDI), the closest relative of ERp29, consistent with a role in secretory protein production. However strikingly different ERp29/PDI ratios were observed in various cell types, suggesting independent regulation and functional roles. Together, these findings associate ERp29 primarily with the early stages of secretory protein production and implicate ERp29 in a distinct functional role that is utilized in most cells. Our identification of several ERp29-enriched cell types suggests a potential selectivity of ERp29 for non-collagenous substrates and provides a physiological foundation for future investigations. PMID- 11897810 TI - Immunohistochemical study of osteopontin expression during distraction osteogenesis in the rat. AB - Distraction osteogenesis (DO) is a limb-lengthening procedure that combines mechanical tension stress with fracture healing to provide a unique opportunity for detailed histological examination of bone formation. Osteopontin (OPN) is a multifunctional matricellular protein believed to play a key role in wound healing and cellular response to mechanical stress. We studied the expression of OPN during DO using standard immunohistochemical (IHC) staining techniques. In addition, we compared the expression of OPN to proliferation (PCNA-positive cells) in the DO gap. After 14 days of distraction in the rat, these stains revealed variations in OPN expression and its relationship to proliferation according to the cell type, tissue type, and mode of ossification examined. Fibroblast-like cells within the central fibrous area exhibited intermittent low levels of OPN, but no relationship was observed between OPN and proliferation. In areas of transchondral ossification, OPN expression was very high in the morphologically intermediate oval cells. During intramembranous ossification, osteoblasts appeared to exhibit a bimodal expression of OPN. Specifically, proliferating pre-osteoblasts expressed osteopontin, but OPN was not detected in the post-proliferative pre-osteoblasts/osteoblasts that border the new bone columns. Finally, intracellular OPN was detected in virtually all of the mature osteoblasts/osteocytes within the new bone columns, while detection of OPN in the matrix of the developing bone columns may increase with the maturity of the new bone. These results imply that the expression of OPN during DO may be more similar to that seen during embryogenesis than would be expected from other studies. Furthermore, the biphasic expression of OPN during intramembranous ossification may exemplify the protein's multi-functional role. Early expression may facilitate pre-osteoblastic proliferation and migration, while the latter downregulation may be necessary for hydroxyapatite crystal formation. PMID- 11897811 TI - Triple immunofluorescence staining with antibodies raised in the same species to study the complex innervation pattern of intrapulmonary chemoreceptors. AB - A general problem in immunocytochemistry is the development of a reliable multiple immunolabeling method when primary antibodies must be used that originate in the same species. We have developed a protocol for the immunodetection of three antigens in a single tissue preparation, using unconjugated primary antibodies raised in the same species. Immunocytochemical detection of neuronal nitric oxide synthase, calcitonin gene-related peptide, and calbindin D28k in the lung of rats demonstrated that part of the pulmonary neuroepithelial bodies are selectively contacted by at least three different nerve fiber populations. The first antigen was detected using tyramide signal amplification, a very sensitive method allowing a dilution of the first primary antibody far beyond the detection limit of fluorescently labeled secondary antibodies. The second antigen was visualized by a fluorophore-conjugated secondary monovalent Fab antibody that at the same time blocks the access of the third secondary antibody to the second primary antibody. Moreover, the monovalence of the Fab fragment prevents the third primary antibody from binding with the second-step secondary antibody. The triple staining technique described here is generally applicable, uses commercially available products only, and allows the detection of three antigens in the same preparation with primary antibodies that are raised in the same species. PMID- 11897812 TI - CP27 localization in the dental lamina basement membrane and in the stellate reticulum of developing teeth. AB - cp27 is a novel gene involved in early vertebrate development that features a distinct protein localization pattern in developing tooth organs. During initial tooth development, CP27 was detected at the epithelial-mesenchymal interface of dental lamina stage tooth organs. At later stages of tooth development, CP27 was localized in the stellate reticulum, the oral mucosa mesenchyme, and alveolar bone. The significant changes in the highly restricted distribution pattern suggest that CP27 might be involved at several different levels during tooth development. PMID- 11897813 TI - Human gene mutations causing infertility. AB - The identification of gene mutations causing infertility in humans remains noticeably deficient at present. Although most males and females with infertility display normal pubertal development, nearly all of the gene mutations in humans have been characterised in people with deficient puberty and subsequent infertility. Gene mutations are arbitrarily categorised into four different compartments (I, hypothalamic; II, pituitary; III, gonadal; and IV, outflow tract). Diagnoses of infertility include hypogonadotrophic hypogonadism (compartments I and II), hypergonadotrophic hypogonadism (III), and obstructive disorders (compartment IV). Most gene mutations identified to date affect gonadal function, but it is also apparent that a large number of important genes in normal fertility have yet to be realised. PMID- 11897816 TI - Genes versus environment in ocular refraction. PMID- 11897815 TI - Organisation of the pericentromeric region of chromosome 15: at least four partial gene copies are amplified in patients with a proximal duplication of 15q. AB - Clinical cytogenetic laboratories frequently identify an apparent duplication of proximal 15q that does not involve probes within the PWS/AS critical region and is not associated with any consistent phenotype. Previous mapping data placed several pseudogenes, NF1, IgH D/V, and GABRA5 in the pericentromeric region of proximal 15q. Recent studies have shown that these pseudogene sequences have increased copy numbers in subjects with apparent duplications of proximal 15q. To determine the extent of variation in a control population, we analysed NF1 and IgH D pseudogene copy number in interphase nuclei from 20 cytogenetically normal subjects by FISH. Both loci are polymorphic in controls, ranging from 1-4 signals for NF1 and 1-3 signals for IgH D. Eight subjects with apparent duplications, examined by the same method, showed significantly increased NF1 copy number (5-10 signals). IgH D copy number was also increased in 6/8 of these patients (4-9 signals). We identified a fourth pseudogene, BCL8A, which maps to the pericentromeric region and is coamplified along with the NF1 sequences. Interphase FISH ordering experiments show that IgH D lies closest to the centromere, while BCL8A is the most distal locus in this pseudogene array; the total size of the amplicon is estimated at approximately 1 Mb. The duplicated chromosome was inherited from either sex parent, indicating no parent of origin effect, and no consistent phenotype was present. FISH analysis with one or more of these probes is therefore useful in discriminating polymorphic amplification of proximal pseudogene sequences from clinically significant duplications of 15q. PMID- 11897818 TI - ARC syndrome is not so rare. PMID- 11897814 TI - Leber hereditary optic neuropathy. AB - Leber hereditary optic neuropathy (LHON) is a mitochondrial genetic disease that preferentially causes blindness in young adult males, affecting about 1 in 25 000 of the British population. It is characterised by bilateral subacute loss of central vision owing to focal degeneration of the retinal ganglion cell layer and optic nerve. Over 95% of LHON cases are primarily the result of one of three mitochondrial DNA (mtDNA) point mutations, G3460A, G11778A, and T14484C, which all involve genes encoding complex I subunits of the respiratory chain. An intriguing feature of LHON is that only approximately 50% of males and approximately 10% of females who harbour a pathogenic mtDNA mutation actually develop the optic neuropathy. This marked incomplete penetrance and gender bias imply that additional mitochondrial and/or nuclear genetic factors must be modulating the phenotypic expression of LHON. It is also likely that environmental factors contribute to the onset of visual failure. However, these secondary precipitating factors remain poorly defined at present. In this review, we describe the natural history of this optic nerve disorder and highlight issues relating to clinical diagnosis, management, and genetic counselling. We also discuss the findings of recently published studies and the light they shed on the complex aetiology and pathophysiology of LHON. PMID- 11897819 TI - Omphalocele in three generations with autosomal dominant transmission. AB - We report a family with nine subjects over three generations affected with an omphalocele requiring surgical intervention within the first few days of life. Because of the vertical transmission and male to male inheritance in our family, we conclude that an autosomal dominant gene caused the omphalocele in the affected family members. The paternal great grandfather of the proband was not clinically affected but produced two children with omphaloceles with different spouses. PMID- 11897817 TI - Prevalence of SDHB, SDHC, and SDHD germline mutations in clinic patients with head and neck paragangliomas. AB - BACKGROUND: Paragangliomas are rare and highly heritable tumours of neuroectodermal origin that often develop in the head and neck region. Germline mutations in the mitochondrial complex II genes, SDHB, SDHC, and SDHD, cause hereditary paraganglioma (PGL). METHODS: We assessed the frequency of SDHB, SDHC, and SDHD gene mutations by PCR amplification and sequencing in a set of head and neck paraganglioma patients who were previously managed in two otolaryngology clinics in the USA. RESULTS: Fifty-five subjects were grouped into 10 families and 37 non-familial cases. Five of the non-familial cases had multiple tumours. Germline SDHD mutations were identified in five of 10 (50%) familial and two of 37 ( approximately 5%) non-familial cases. R38X, P81L, H102L, Q109X, and L128fsX134 mutations were identified in the familial cases and P81L was identified in the non-familial cases. Both non-familial cases had multiple tumours. P81L and R38X mutations have previously been reported in other PGL families and P81L was suggested as a founder mutation. Allelic analyses of different chromosomes carrying these mutations did not show common disease haplotypes, strongly suggesting that R38X and P81L are potentially recurrent mutations. Germline SDHB mutations were identified in two of 10 (20%) familial and one of 33 ( approximately 3%) non-familial cases. P131R and M71fsX80 were identified in the familial cases and Q59X was identified in the one non-familial case. The non-familial case had a solitary tumour. No mutations could be identified in the SDHC gene in the remaining four families and 20 sporadic cases. CONCLUSIONS: Mutations in SDHD are the leading cause of head and neck paragangliomas in this clinic patient series. SDHD and SDHB mutations account for 70% of familial cases and approximately 8% of non-familial cases. These results also suggest that the commonness of the SDHD P81L mutation in North America is the result of both a founder effect and recurrent mutations. PMID- 11897820 TI - A novel locus for brachydactyly type A1 on chromosome 5p13.3-p13.2. PMID- 11897821 TI - A Val227Ala polymorphism in the peroxisome proliferator activated receptor alpha (PPARalpha) gene is associated with variations in serum lipid levels. PMID- 11897822 TI - ATM mutations in Finnish breast cancer patients. PMID- 11897823 TI - A single base alteration in the CGG repeat region of FMR1: possible effects on gene expression and phenotype. PMID- 11897825 TI - Prader-Willi syndrome and a deletion/duplication within the 15q11-q13 region. PMID- 11897824 TI - Mosaicism for FMR1 and FMR2 deletion: a new case. PMID- 11897826 TI - The frequency of mtDNA 8994 polymorphism and detection of the NARP 8993 mutation. PMID- 11897827 TI - An aetiological study of 25 mentally retarded adults with autism. PMID- 11897828 TI - Familial pericentric inversion of chromosome 5 in a family with benign neonatal convulsions. PMID- 11897829 TI - Phenotypic effects of mosaicism for a 47,XXX cell line in Turner syndrome. PMID- 11897830 TI - Dilated cardiomyopathy, sudden cardiac death, hypoplastic discs, and retinal detachment: a new autosomal dominant syndrome. PMID- 11897831 TI - Hirschsprung disease and L1CAM: is the disturbed sex ratio caused by L1CAM mutations? PMID- 11897832 TI - Contribution of BRCA2 germline mutations to hereditary breast/ovarian cancer in Germany. PMID- 11897833 TI - Investigation of the GRB2, GRB7, and CSH1 genes as candidates for the Silver Russell syndrome (SRS) on chromosome 17q. PMID- 11897834 TI - Human T and risk for neural tube defects. PMID- 11897835 TI - Reproductive counselling for women with myotonic dystrophy. PMID- 11897836 TI - The TREK two P domain K+ channels. PMID- 11897837 TI - Heterogeneous expression of KChIP2 isoforms in the ferret heart. AB - Kv4 channels are believed to underlie the rapidly recovering cardiac transient outward current (I(to)) phenotype. However, heterologously expressed Kv4 channels fail to fully reconstitute the native current. Kv channel interacting proteins (KChIPs) have been shown to modulate Kv4 channel function. To determine the potential involvement of KChIPs in the rapidly recovering I(to), we cloned three KChIP2 isoforms (designated fKChIP2, 2a and 2b) from the ferret heart. Based upon immunoblot data suggesting the presence of a potential endogenous KChIP-like protein in HEK 293, CHO and COS cells but absence in Xenopus oocytes, we coexpressed Kv4.3 and the fKChIP2 isoforms in Xenopus oocytes. Functional analysis showed that while all fKChIP2 isoforms produced a fourfold acceleration of recovery kinetics compared to Kv4.3 expressed alone, only fKChIP2a produced large depolarizing shifts in the V(1/2) of steady-state activation and inactivation as seen for the native rapidly recovering I(to). Analysis of RNA and protein expression of the three fKChIP2 isoforms in ferret ventricles showed that fKChIP2b was most abundant and was expressed in a gradient paralleling the rapidly recovering I(to) distribution. Ferret KChIP2 and 2a were expressed at very low levels. The ventricular expression distribution suggests that fKChIP2 isoforms are involved in modulation of the rapidly recovering I(to); however, additional regulatory factors are also likely to be involved in generating the native current. PMID- 11897838 TI - Expression pattern and functional characteristics of two novel splice variants of the two-pore-domain potassium channel TREK-2. AB - Two novel alternatively spliced isoforms of the human two-pore-domain potassium channel TREK-2 were isolated from cDNA libraries of human kidney and fetal brain. The cDNAs of 2438 base pairs (bp) (TREK-2b) and 2559 bp (TREK-2c) encode proteins of 508 amino acids each. RT-PCR showed that TREK-2b is strongly expressed in kidney (primarily in the proximal tubule) and pancreas, whereas TREK-2c is abundantly expressed in brain. In situ hybridization revealed a very distinct expression pattern of TREK-2c in rat brain which partially overlapped with that of TREK-1. Expression of TREK-2b and TREK-2c in human embryonic kidney (HEK) 293 cells showed that their single-channel characteristics were similar. The slope conductance at negative potentials was 163 +/- 5 pS for TREK-2b and 179 +/- 17 pS for TREK-2c. The mean open and closed times of TREK-2b at -84 mV were 133 +/- 16 and 109 +/- 11 micros, respectively. Application of forskolin decreased the whole cell current carried by TREK-2b and TREK-2c. The sensitivity to forskolin was abolished by mutating a protein kinase A phosphorylation site at position 364 of TREK-2c (construct S364A). Activation of protein kinase C (PKC) by application of phorbol-12-myristate-13-acetate (PMA) also reduced whole-cell current. However, removal of the putative TREK-2b-specific PKC phosphorylation site (construct T7A) did not affect inhibition by PMA. Our results suggest that alternative splicing of TREK-2 contributes to the diversity of two-pore-domain K+ channels. PMID- 11897839 TI - Functional differences between alpha subunit isoforms of the rat Na,K-ATPase expressed in Xenopus oocytes. AB - The functional properties of the three most widely distributed alpha subunit isoforms of the Na,K-ATPase are not well known, particularly concerning the voltage dependence of their activity and cation binding kinetics. We measured the electrogenic activity generated by Na,K-ATPases resulting from co-expression of the rat alpha1, alpha2* or alpha3* subunits with the rat beta1 subunit in Xenopus oocytes; alpha2* and alpha3* are ouabain-resistant mutants of the alpha2 and alpha3 isoform, which allowed selective inhibition of the endogenous Na(+),K(+) pump of the oocyte. In oocytes expressing the three isoforms of the alpha subunit, K(+) induced robust outward currents that were largely ouabain sensitive. In addition, ouabain-sensitive inward currents were recorded for all three isoforms in sodium-free and potassium-free acid solutions. The very similar voltage dependence of the Na(+),K(+)-pump activity observed in the absence of extracellular Na(+) indicated a similar stoichiometry of the transported cations by the three isoforms. The affinity for extracellular K(+) was slightly lower for the alpha2* and alpha3* than for the alpha1 isoform. The alpha2* isoform was, however, more sensitive to voltage-dependent inhibition by extracellular Na(+), indicating a higher affinity of the extracellular Na(+) site in this isoform. We measured and controlled [Na(+)](i) using a co-expressed amiloride-sensitive Na(+) channel. The intracellular affinity for Na(+) was slightly higher in the alpha2* than in the alpha1 or alpha3* isoforms. These results suggest that the alpha2 isoform could have an activity that is strongly dependent upon [Na(+)](o) and [K(+)](o). These concentrations could selectively modulate its activity when large variations are present, for instance in the narrow intercellular spaces of brain or muscle tissues. PMID- 11897840 TI - Alpha(1H) mRNA in single skeletal muscle fibres accounts for T-type calcium current transient expression during fetal development in mice. AB - Calcium channels are essential for excitation-contraction coupling and muscle development. At the end of fetal life, two types of Ca(2+) currents can be recorded in muscle cells. Whereas L-type Ca(2+) channels have been extensively studied, T-type channels have been poorly characterized in skeletal muscle. We describe here the functional and molecular properties of T-type calcium channels in developing mouse skeletal muscle. The T-type current density increased transiently during prenatal myogenesis with a maximum at embryonic day E16 followed by a drastic decrease until birth. This current showed similar electrophysiological and pharmacological properties at all examined stages. It displayed a wide window current centred at about -35 and -55 mV in 10 and 2 mM external Ca(2+), respectively. Activation and inactivation kinetics were fast (3 and 16 ms, respectively). The current was inhibited by nickel and amiloride with an IC(50) of 5.4 and 156 microM, respectively, values similar to those described for cloned T-type alpha(1H) channels. Whole muscle tissue RT-PCR analysis revealed mRNAs corresponding to alpha(1H) and alpha(1G) subunits in the fetus but not in the adult. However, single-fibre RT-PCR demonstrated that only alpha(1H) mRNA was present in prenatal fibres, suggesting that the alpha(1G) transcript present in muscle tissue must be expressed by non-skeletal muscle cells. Altogether, these results demonstrate that the alpha(1H) subunit generates functional T-type calcium channels in developing skeletal muscle fibres and suggest that these channels are involved in the early stages of muscle differentiation. PMID- 11897841 TI - Evidence for two distinct processes in the final stages of neurotransmitter release as detected by binomial analysis in calcium and strontium solutions. AB - The statistical parameters underlying acetylcholine (ACh) release were studied using Ca(2+) and Sr(2+) ions to promote ACh secretion. Experiments were performed at frog neuromuscular junctions using electrophysiological recording techniques. Increases in asynchronous ACh release, reflected as the frequency of occurrence of miniature end-plate potentials (MEPP(f)), were evoked by high potassium depolarization in either Ca(2+) or Sr(2+) solutions. Increases in MEPP(f) mediated by Ca(2+) were of very low probability and well-described by a Poisson distribution whilst similar MEPP(f) increases mediated by Sr(2+) were best described as a simple binomial distribution. From the binomial distribution in Sr(2+) solutions, values for the average probability of release (p) and the number of releasable ACh quanta (n) may be determined (whereby mean MEPP(f) = np). In Sr(2+) solutions, values of p were independent of both bin width and of the value of n, suggesting that both n and p were stationary. Calculations of p using the simple binomial distribution in Sr(2+) solutions gave theoretical values for the third moment of the mean which were indistinguishable from the experimental distribution. These results, in conjunction with Monte Carlo simulations of the data, suggest that spatial and temporal variance do not measurably affect the analysis. Synchronous ACh release evoked by nerve impulses (end-plate potentials, EPPs) follow a simple binomial distribution in both Ca(2+) and Sr(2+) solutions. Similar mean levels of synchronous ACh release (m, where m = np) were produced by lower values of p and higher values of n in Ca(2+) as compared to Sr(2+). The statistical analyses suggest the presence of two different Ca(2+)-dependent steps in the final stages of neurotransmitter release. The results are discussed in accordance with (i) statistical models for quantal neurotransmitter release, (ii) the role of Sr(2+) as a partial agonist for evoked ACh release, and (iii) the specific loci that may represent the sites of Ca(2+) and Sr(2+) sensitivity. PMID- 11897842 TI - Selective reduction by dopamine of excitatory synaptic inputs to pyramidal neurons in primate prefrontal cortex. AB - We have employed in vitro physiological methods to investigate dopaminergic modulation of excitatory synaptic transmission in monkey prefrontal cortex (PFC) circuits. We show that combined activation of D1-like and D2-like dopamine receptors results in the reduction of extracellular stimulation-evoked isolated EPSCs in layer 3 pyramidal neurons. Using paired recordings from synaptically connected pyramidal neurons we have determined the basic properties of unitary synaptic connections between layer 3 pyramids in the primate PFC and, interestingly, we found that dopamine does not reduce synaptic transmission between nearby pairs of synaptically coupled PFC pyramidal neurons. This input specificity may be a critical aspect of the dopaminergic regulation of recurrent excitatory circuits in the PFC. PMID- 11897843 TI - Ultrastructural basis of synaptic transmission between endbulbs of Held and bushy cells in the rat cochlear nucleus. AB - Auditory nerve fibres make large excitatory synaptic contacts, the endbulbs of Held, with bushy cells in the anteroventral cochlear nucleus (AVCN). We have used serial-section electron microscopy to reconstruct seven endbulbs of Held in contact with three different AVCN bushy cells from a 25-day-old rat, as a basis for interpreting our previous physiological results at this connection. Four endbulbs of Held contacting the same bushy cell were completely reconstructed. The number of separate synaptic specializations within these endbulbs varied from 85 to 217, with a mean of 155. Detailed measurements were obtained from high magnification segments of four endbulbs contacting three different bushy cells. Large variability was found in the size of synaptic specializations within an individual endbulb. The size of postsynaptic densities (PSDs) varied between endbulbs (mean PSD area 0.03, 0.07, 0.07 and 0.18 microm(2); n = 4 endbulbs). The number of morphologically docked vesicles at individual specializations within the same endbulb varied considerably (between 1 and 102). The mean number of morphologically docked vesicles per specialization differed between endbulbs (mean numbers of docked vesicles per specialization = 2.1, 3.7, 5.3, 14.8; n = 4 endbulbs). Despite these large differences, the density of docked vesicles per square micron of PSD was similar between endbulbs (54, 80, 81, 83 docked vesicles per microm(2); n = 4 endbulbs). Within an endbulb, a linear relationship was found between the number of docked vesicles and PSD area, and between PSD area and the number of undocked vesicles within 150 nm of the active zone. The ratio of undocked vesicles (< 150 nm) to docked vesicles ranged from 2 to 5 in different endbulbs (n = 4 endbulbs). These structural observations are discussed in relation to the functional properties of synaptic transmission between endbulbs of Held and bushy cells in the AVCN. PMID- 11897844 TI - External anions and cations distinguish between AMPA and kainate receptor gating mechanisms. AB - Experiments were designed to examine if ion-flow through alpha-amino-3-hydroxy-5 methyl-isoxazolepropionic acid (AMPA) or kainate receptors interferes with protein structures associated with the gating machinery. Gating was studied using ultra-fast drug perfusion of outside-out patches containing rat GluR-A or GluR6 subunits excised from transfected human embryonic kidney cells. Deactivation rates of GluR6 kainate receptors observed following brief L-glutamate (10 mM Glu, 1 ms) applications differed by two to threefold in high (405 mM symmetrical Na(+), tau(decay) = 2.7 ms at -100 mV) and low ionic strength (55 mM, tau(decay) = 1.1 ms) solutions. In comparison, GluR-A AMPA receptors were much less sensitive. Ion effects on GluR6 receptors did not reflect surface potential screening or ion-agonist competition at the agonist-binding site since deactivation rates were slower in high ionic strength solutions. Moreover, the apparent agonist affinity did not decrease with increasing ionic strength (e.g. 55 mM, EC(50) = 110 microM vs. 405 mM, EC(50) = 61 microM). GluR6 responses were strongly dependent on ions present on the external, but not the internal, side of the plasma membrane. Decay kinetics was regulated by the type of ion present suggesting that the chemical nature of the solution, not its ionic strength, governed channel behaviour. Both external anions and cations modulated the amplitude and decay kinetics of GluR6 responses in a concomitant manner. AMPA receptor responses recorded in identical ionic conditions did not exhibit this behaviour. These results identify a novel mechanism that distinguishes AMPA and kainate receptors. External ions regulate the gating machinery of kainate receptors through an allosteric mechanism that involves both anions and cations. PMID- 11897845 TI - Reduced NR2A expression and prolonged decay of NMDA receptor-mediated synaptic current in rat vagal motoneurons following axotomy. AB - To elucidate characteristic changes in the N-methyl-D-aspartate (NMDA) receptor on neurons following axotomy, subunit expressions and functional features of the NMDA receptor were examined in the dorsal motor nucleus of vagus (DMV) of rats receiving vagal axotomy at the neck. Western blotting analysis demonstrated that the expression of NR2A decreased 2-3 days after in vivo axotomy, while expression of NR1 and NR2B, NR2C and NR2D subunits did not change significantly. To examine the functional changes, patch clamp recordings in whole-cell mode were employed on the axotomized DMV neurons identified by retrograde labelling with fluorescent dye. The amplitude ratios of ifenprodil-sensitive components of NMDA response and D,L-2-amino-5-phosphovaleric acid (APV)-sensitive evoked postsynaptic current increased after axotomy. In addition, APV-sensitive postsynaptic currents exhibited a longer decay time in identified axotomized vagal motoneurons than in control neurons. No significant differences in the current density of the NMDA response and the peak amplitude of APV-sensitive synaptic currents were observed between axotomized and intact DMV neurons. In conclusion, a decrease in NR2A expression results in the appearance of functional characteristics of the NMDA receptor predominantly containing the NR2B subunit. This might lead to a long term increase of the susceptibility of neurons to excitotoxicity. PMID- 11897846 TI - Slow removal of Na(+) channel inactivation underlies the temporal filtering property in the teleost thalamic neurons. AB - It has been previously shown that the "large cell" in the corpus glomerulosum (CG) of a teleost brain has a low-pass temporal filtering property. It fires a single spike only in response to temporally sparse synaptic inputs and thus extracts temporal aspects of afferent activities. To explore the ionic mechanisms underlying this property, we quantitatively studied voltage-gated Na(+) channels of the large cell in the CG slice preparation of the marine filefish by means of whole-cell patch clamp recordings in the voltage-clamp mode. Recorded Na(+) current was well described using the Hodgkin-Huxley "m(3)h" model. It was revealed that the Na(+) channels have a novel feature: remarkably slow recovery from inactivation. In other words, the time constant for the "h" gate was extremely large (approximately 100 ms at -80 to -50 mV). In order to test whether the analysed Na(+) current serves as a mechanism for filtering, the behaviour of the membrane model incorporating the Na(+) channel was simulated using a computer program called NEURON. In response to current injections, the membrane model displayed low-pass filtering and firing properties similar to those reported in real cells. The present results suggest that slow removal of Na(+) channel inactivation serves as a crucial mechanism for the low-pass temporal filtering property of the large cell. The simulation study also suggested that velocity and/or amplitude of a spike propagating though an axon expressing Na(+) channels of this type could potentially be modulated depending on the preceding activities of the cells. PMID- 11897847 TI - "Sleepy" inward rectifier channels in guinea-pig cardiomyocytes are activated only during strong hyperpolarization. AB - K(+) channels of isolated guinea-pig cardiomyocytes were studied using the patch clamp technique. At transmembrane potentials between -120 and -220 mV we observed inward currents through an apparently novel channel. The novel channel was strongly rectifying, no outward currents could be recorded. Between -200 and -160 mV it had a slope conductance of 42.8 +/- 3.0 pS (S.D.; n = 96). The open probability (P(o)) showed a sigmoid voltage dependence and reached a maximum of 0.93 at -200 mV, half-maximal activation was approximately -150 mV. The voltage dependence of P(o) was not affected by application of 50 microM isoproterenol. The open-time distribution could be described by a single exponential function, the mean open time ranged between 73.5 ms at -220 mV and 1.4 ms at -160 mV. At least two exponential components were required to fit the closed time distribution. Experiments with different external Na(+), K(+) and Cl(-) concentrations suggested that the novel channel is K(+) selective. Extracellular Ba(2+) ions gave rise to a voltage-dependent reduction in P(o) by inducing long closed states; Cs(+) markedly reduced mean open time at -200 mV. In cell-attached recordings the novel channel frequently converted to a classical inward rectifier channel, and vice versa. This conversion was not voltage dependent. After excision of the patch, the novel channel always converted to a classical inward rectifier channel within 0-3 min. This conversion was not affected by intracellular Mg(2+), phosphatidylinositol (4,5)-bisphosphate or spermine. Taken together, our findings suggest that the novel K(+) channel represents a different "mode" of the classical inward rectifier channel in which opening occurs only at very negative potentials. PMID- 11897848 TI - Effects of phosphocreatine on SR Ca(2+) regulation in isolated saponin permeabilized rat cardiac myocytes. AB - The effects of phosphocreatine (PCr) on sarcoplasmic reticulum (SR) Ca(2+) regulation were investigated in saponin-permeabilized rat ventricular myocytes. Cells were perfused continuously with weakly Ca(2+)-buffered solutions approximating to the intracellular milieu. Ca(2+) release from the SR was detected using Fura-2 or Fluo-3. Withdrawal of PCr reduced the frequency of spontaneous Ca(2+) release by 12.8 +/- 3.4 % (n = 9) and the amplitude of the spontaneous Ca(2+) transient by 17.4 +/- 3.1 % (n = 9). Stepwise reductions in [PCr] progressively increased the time for the spontaneous Ca(2+) transient to rise from 25 to 100 % of the maximum value (TP75) and to fall by 75 % of the peak level (DT75). Following complete PCr withdrawal, the TP75 and the DT75 were 147.1 +/- 13.2 and 174.8 +/- 23.2 % of the control values, respectively. Experiments involving confocal microscopy showed that PCr withdrawal decreased the propagation velocity of spontaneous Ca(2+) waves. PCr withdrawal also reduced the frequency and amplitude, but increased the duration of spontaneous Ca(2+) sparks. Rapid application of 20 mM caffeine was used to assess the SR Ca(2+) content at the point of spontaneous Ca(2+) release. In the absence of PCr, the amplitude of the caffeine-induced Ca(2+) transient was 18.4 +/- 2.7 % (n = 9) lower than in the presence of 10 mM PCr. This suggests that PCr withdrawal reduces the maximum SR Ca(2+) content that can be sustained before spontaneous Ca(2+) release occurs. These results suggest that local ADP buffering by PCr is essential for normal Ca(2+) regulation by the SR. Prolongation of the descending phase of the spontaneous Ca(2+) transient is consistent with a reduction in the efficiency of the SR Ca(2+) pump due to ADP accumulation. The fact that spontaneous Ca(2+) release occurs at a lower SR Ca(2+) content in the absence of PCr suggests that the Ca(2+) release mechanism may also be affected. These effects may be of relevance in circumstances where PCr depletion and Ca(2+) overload occur, such as myocardial ischaemia or anoxia. PMID- 11897849 TI - Modulatory role of focal adhesion kinase in regulating human pulmonary arterial endothelial barrier function. AB - The adhesive force generated by the interaction of integrin receptors with extracellular matrix (ECM) at the focal adhesion complex may regulate endothelial cell shape, and thereby the endothelial barrier function. We studied the role of focal adhesion kinase (FAK) activated by integrin signalling in regulating cell shape using cultured human pulmonary artery endothelial cells. We used FAK antisense oligonucleotide (targeted to the 3'-untranslated region of FAK mRNA (5' CTCTGGTTGATGGGATTG-3') to determine the role of FAK in the mechanism of thrombin induced increase in endothelial permeability. Reduction in FAK expression by the antisense augmented the thrombin-induced decrease in transendothelial electrical resistance (decrease in mock transfected cells of -43 +/- 1 % and in sense transfected cells of -40 +/- 4 %, compared to the decrease in antisense transfected cells of -60 +/- 3 %). Reduction in FAK expression also prolonged the drop in electrical resistance and prevented the recovery seen in control endothelial cells. Thus, the thrombin-induced increase in permeability is both greater and attenuated in the absence of FAK expression. Inhibition of actin polymerization with latrunculin-A prevented the translocation of FAK to focal adhesion sites and tyrosine phosphorylation of FAK and paxillin, and concomitantly reduced the thrombin-induced decrease in electrical resistance by approximately 50 %. Thus, the modulatory role of FAK on endothelial barrier function is dependent on actin polymerization. FAK translocation to focal adhesion complex in endothelial cells guided by actin cables and the consequent activation of FAK-associated proteins serve to reverse the decrease in endothelial barrier function caused by inflammatory mediators such as thrombin. PMID- 11897850 TI - MgATP counteracts intracellular proton inhibition of the sodium-calcium exchanger in dialysed squid axons. AB - Intracellular Na(+) and H(+) inhibit Na(+)-Ca(2+) exchange. ATP regulates exchange activity by altering kinetic parameters for Ca(2+)(i), Na(+)(i) and Na(+)(o). The role of the Ca(2+)(i)regulatory site on Na(+)(i)-H(+)(i)-ATP interactions was explored by measuring the Na(+)(o)-dependent (45)Ca(2+) efflux (Na(+)(o)-Ca(2+)(i) exchange) and Ca(2+)(i)-dependent (22)Na(+) efflux (Na(+)(o) Na(+)(i) exchange) in intracellular-dialysed squid axons. Our results show that: (1) without ATP, inhibition by Na(+)(i) is strongly dependent on H(+)(i). Lowering the pH(i) by 0.4 units from its physiological value of 7.3 causes 80 % inhibition of Na(+)(o)-Ca(2+)(i) exchange; (2) in the presence of MgATP, H(+)(i) and Na(+)(i) inhibition is markedly diminished; and (3) experiments on Na(+)(o) Na(+)(i) exchange indicate that the drastic changes in the Na(+)(i)-H(+)(i)-ATP interactions take place at the Ca(2+)(i) regulatory site. The increase in Ca(2+)(i) affinity induced by ATP at acid pH (6.9) can be mimicked by a rise in pH(i) from 6.9 to 7.3 in the absence of the nucleotide. We conclude that ATP modulation of the Na(+)-Ca(2+) exchange occurs by protection from intracellular proton and sodium inhibition. These findings are predicted by a model where: (i) the binding of Ca(2+) to the regulatory site is essential for translocation but not for the binding of Na(+)(i) or Ca(2+)(i) to the transporting site; (ii) H(+)(i) competes with Ca(2+)(i) for the same form of the exchanger without an effect on the Ca(2+)(i) transporting site; (iii) protonation of the carrier increases the apparent affinity and changes the cooperativity for Na(+)(i) binding; and (iv) ATP prevents both H(+)(i) and Na(+)(i)-effects. The relief of H(+) and Na(+) inhibition induced by ATP could be important in cardiac ischaemia, in which a combination of acidosis and rise in [Na(+)](i) occurs. PMID- 11897851 TI - Multiple regulation by external ATP of nifedipine-insensitive, high voltage activated Ca(2+) current in guinea-pig mesenteric terminal arteriole. AB - We investigated the receptor-mediated regulation of nifedipine-insensitive, high voltage-activated Ca(2+) currents in guinea-pig terminal mesenteric arterioles (I(mVDCC)) using the whole-cell clamp technique. Screening of various vasoactive substances revealed that ATP, histamine and substance P exert modulatory effects on I(mVDCC). The effects of ATP on I(mVDCC) after complete P2X receptor desensitization exhibited a complex concentration dependence. With 5 mM Ba(2+), ATP potentiated I(mVDCC) at low concentrations (approximately 1-100 microM), but inhibited it at higher concentrations (>100 microM). The potentiating effects of ATP were abolished by suramin (100 microM) and PPADS (10 microM) and by intracellular application of GDPbetaS (500 microM), whereas a substantial part of I(mVDCC) inhibition by milimolar concentrations of ATP remained unaffected; due probably to its divalent cation chelating actions. In divalent cation-free solution, I(mVDCC) was enlarged and underwent biphasic effects by ATPgammaS and ADP, while 2-methylthio ATP (2MeSATP) exerted only inhibition, and pyrimidines such as UTP and UDP were ineffective. ATP-induced I(mVDCC) potentiation was selectively inhibited by anti-Galpha(s) antibodies or protein kinase A (PKA) inhibitory peptides and mimicked by dibutyryl cAMP. In contrast, ATP-induced inhibition was selectively inhibited by Galpha(q/11) antibodies or protein kinase C (PKC) inhibitory peptides and mimicked by PDBu. Pretreatment with pertussis toxin was ineffective. The apparent efficacy for I(mVDCC) potentiation with PKC inhibitors was: ATPgammaS > ATP>/=ADP and for inhibition with PKA inhibitors was: 2MeSATP > ATPgammaS > ATP > ADP. Neither I(mVDCC) potentiation nor inhibition showed voltage dependence. These results suggest that I(mVDCC) is multi phasically regulated by external ATP via P2Y(11)-resembling receptor/G(s)/PKA pathway, P2Y(1)-like receptor/G(q/11)/PKC pathway, and metal chelation. PMID- 11897853 TI - pH and rate of "dark" events in toad retinal rods: test of a hypothesis on the molecular origin of photoreceptor noise. AB - Thermal activation of the visual pigment constitutes a fundamental constraint on visual sensitivity. Its electrical correlate in the membrane current of dark adapted rods are randomly occurring discrete "dark events" indistinguishable from responses to single photons. It has been proposed that thermal activation occurs in a small subpopulation of rhodopsin molecules where the Schiff base linking the chromophore to the protein part is unprotonated. On this hypothesis, rates of thermal activation should increase strongly with rising pH. The hypothesis has been tested by measuring the effect of pH changes on the frequency of discrete dark events in red rods of the common toad Bufo bufo. Dark noise was recorded from isolated rods using the suction pipette technique. Changes in cytoplasmic pH upon manipulations of extracellular pH were quantified by measuring, using fast single-cell microspectrophotometry, the pH-dependent metarhodopsin I metarhodopsin II equilibrium and subsequent metarhodopsin III formation. These measurements show that, in the conditions of the electrophysiological experiments, changing perfusion pH from 6.5 to 9.3 resulted in a cytoplasmic pH shift from 7.6 to 8.5 that was readily sensed by the rhodopsin. This shift, which implies an 8-fold decrease in cytoplasmic [H(+)], did not increase the rate of dark events. The results contradict the hypothesis that thermal pigment activation depends on prior deprotonation of the Schiff base. PMID- 11897854 TI - Oxidation induces a Cl(-)-dependent cation conductance in human red blood cells. AB - Oxidative stress induces complex alterations of membrane proteins in red blood cells (RBCs) eventually leading to haemolysis. To study changes of membrane ion permeability induced by oxidative stress, whole-cell patch-clamp recordings and haemolysis experiments were performed in control and oxidised human RBCs. Control RBCs exhibited a small cation-selective whole-cell conductance (236 +/- 38 pS; n = 8) which was highly sensitive to the external Cl(-) concentration: replacement of NaCl in the bath by sodium gluconate induced an increase of this cation conductance by about 85 %. Exposing RBCs to t-butylhydroxyperoxide (1 mM for 10 min) induced a twofold increase in this cation conductance which was further stimulated after replacement of extracellular Cl(-) by gluconate, Br(-), I(-) or SCN(-). In addition, lowering the ionic strength of the bath solution by isosmotic substitution of NaCl by sorbitol activated the cation conductance. The Cl(-)-sensitive and oxidation-induced cation conductance was Ca(2+) permeable, exhibited a permselectivity of Cs(+) > K(+) > Na(+) = Li(+) >> NMDG(+), and was partially inhibited by amiloride (1 mM) and almost completely inhibited by GdCl(3) (150 microM), but was insensitive to TEA, BaCl(2), NPPB, flufenamic acid or quinidine. DIDS (100 microM) reversibly inhibited the activation of the cation conductance by removal of external Cl(-). Oxidation induced haemolysis in NaCl bathed human RBCs. This haemolysis was attenuated by amiloride (1 mM) and inhibited by replacement of bath Na(+) by the impermeant cation NMDG(+). The Na(+)- and Ca(2+)-permeable conductance might be involved in haemolytic diseases induced by elevated oxidative stress, such as glucose-6-phosphate dehydrogenase deficiency. PMID- 11897852 TI - Four cell types with distinctive membrane properties and morphologies in lamina I of the spinal dorsal horn of the adult rat. AB - Lamina I of the spinal dorsal horn plays an important role in the processing and relay of nociceptive information. Signal processing depends, in part, on neuronal membrane properties. Intrinsic membrane properties of lamina I neurons were therefore investigated using whole cell patch clamp recordings in a slice preparation of adult rat spinal cord. Based on responses to somatic current injection, four cell types were identified: tonic, which fire comparatively slowly but continuously throughout stimulation; phasic, which fire a high frequency burst of variable duration; delayed onset, which fire irregularly and with a marked delay to the first spike; and single spike, which typically fire only one action potential even when strongly depolarised. Classification by spiking pattern was further refined by identification of characteristic stimulus response curves and quantification of several response parameters. Objectivity of the classification was confirmed by cluster analysis. Responses to stimulus trains and synaptic input as well as the kinetics of spontaneous synaptic events revealed differences in the signal processing characteristics of the cell types: tonic and delayed onset cells appeared to act predominantly as integrators whereas phasic and single spike cells acted as coincidence detectors. Intracellular labelling revealed a significant correlation between morphological and physiological cell types: tonic cells were typically fusiform, phasic cells were pyramidal, and delayed onset and single spike cells were multipolar. Thus, there are multiple physiological cells types in lamina I with specific morphological correlates and distinctive signal processing characteristics that confer significant differences in the transduction of input into spike trains. PMID- 11897855 TI - Simultaneous activation of gamma and theta network oscillations in rat hippocampal slice cultures. AB - Hippocampal activity in vivo is characterized by concurrent oscillations at theta (4-15 Hz) and gamma (20-80 Hz) frequencies. Here we show that cholinergic receptor activation (methacholine 10-20 nm) in hippocampal slice cultures induces an oscillatory mode of activity, in which the intrinsic network oscillator (located in the CA3 area) expresses simultaneous theta and gamma network oscillations. Pyramidal cells display synaptic theta oscillations, characterized by cycles consisting of population EPSP-IPSP sequences that are dominated by population IPSPs. These rhythmic IPSPs most probably result from theta-modulated spiking activity of several interneurons. At the same time, the majority of interneurons consistently display synaptic gamma oscillations. These oscillatory cycles consist of fast depolarizing rhythmic events that are likely to reflect excitatory input from CA3 pyramidal cells. Interneurons comprising this functional group were identified morphologically. They include four known types of interneurons (basket, O-LM, bistratified and str. lucidum-specific cells) and one new type of CA3 interneuron (multi-subfield cell). The oscillatory activity of these interneurons is only weakly correlated between neighbouring cells, and in about half of these (44 %) is modulated by depolarizing theta rhythmicity. The overall characteristics of acetylcholine-induced oscillations in slice cultures closely resemble the rhythmicity observed in hippocampal field and single cell recordings in vivo. Both rhythmicities depend on intrinsic synaptic interactions, and are expressed by different cell types. The fact that these oscillations persist in a network lacking extra-hippocampal connections emphasizes the importance of intrinsic mechanisms in determining this form of hippocampal activity. PMID- 11897857 TI - Hydrogen peroxide increases depolarization-induced contraction of mechanically skinned slow twitch fibres from rat skeletal muscles. AB - The effect of exogenous hydrogen peroxide (H(2)O(2)) on excitation-contraction (E C) coupling and sarcoplasmic reticulum (SR) function was compared in mechanically skinned slow twitch fibres (prepared from the soleus muscles) and fast twitch fibres (prepared from the extensor digitorum longus; EDL muscles) of adult rats. Equilibration (5 min) with 1 mM H(2)O(2) diminished the ability of the Ca(2+) depleted SR to reload Ca(2+) in both slow (P < 0.01) and fast twitch fibres (P < 0.05) compared to control. Under conditions when all Ca(2+) uptake was prevented, 1 mM H(2)O(2) increased SR Ca(2+) "leak" in fast twitch fibres by 24 +/- 5 % (P < 0.05), but leak was not altered in slow twitch fibres. Treatment with 1 mM H(2)O(2) also increased the peak force of low [caffeine] contracture by approximately 45% in both fibre types compared to control (P < 0.01), which could be partly reversed following treatment with 10 mM dithiothreitol (DTT). The changes in SR function caused by 1 mM H(2)O(2) were associated with an approximately 65% increase in the peak height of depolarization-induced contractile response (DICR) in slow twitch fibres, compared to control (no H(2)O(2); P < 0.05). In contrast, peak contractile force of fast twitch fibres was not altered by 1 mM H(2)O(2) treatment. Equilibration with 5 mM H(2)O(2) induced a spontaneous force response in both slow and fast twitch fibres, which could be partly reversed by 2 min treatment with 10 mM DTT. Peak DICR was also increased approximately 40% by 5 mM H(2)O(2) in slow twitch fibres compared to control (no H(2)O(2); P < 0.05). Our results indicate that exogenous H(2)O(2) increases depolarization-induced contraction of mechanically skinned slow but not fast twitch fibres. The increase in depolarization-induced contraction in slow twitch fibres might be mediated by an increased SR Ca(2+) release during contraction and/or an increase in Ca(2+) sensitivity. PMID- 11897856 TI - Differential effects of sarcoplasmic reticular Ca(2+)-ATPase inhibition on charge movements and calcium transients in intact amphibian skeletal muscle fibres. AB - A hypothesis in which intramembrane charge reflects a voltage sensing process allosterically coupled to transitions in ryanodine receptor (RyR)-Ca(2+) release channels as opposed to one driven by release of intracellularly stored Ca(2+) would predict that such charging phenomena should persist in skeletal muscle fibres unable to release stored Ca(2+). Charge movement components were accordingly investigated in intact voltage-clamped amphibian fibres treated with known sarcoplasmic reticular (SR) Ca(2+)-ATPase inhibitors. Cyclopiazonic acid (CPA) pretreatment abolished Ca(2+) transients in fluo-3-loaded fibres following even prolonged applications of caffeine (10 mM) or K(+) (122 mM). Both CPA and thapsigargin (TG) transformed charge movements that included delayed (q(gamma)) "hump" components into simpler decays. However, steady-state charge-voltage characteristics were conserved to values (maximum charge, Q(max) approximately equal to 20-25 nC microF(-1); transition voltage, V* approximately equal to -40 to-50 mV; steepness factor, k approximately equal to 6-9 mV; holding voltage -90 mV) indicating persistent q(gamma) charge. The features of charge inactivation similarly suggested persistent q(beta) and q(gamma) charge contributions in CPA treated fibres. Perchlorate (8.0 mM) restored the delayed kinetics shown by "on" q(gamma) charge movements, prolonged their "off" decays, conserved both Q(max) and k, yet failed to restore the capacity of such CPA-treated fibres for Ca(2+) release. Introduction of perchlorate (8.0 mM) or caffeine (0.2 mM) to tetracaine (2.0 mM)-treated fibres, also known to restore q(gamma) charge, similarly failed to restore Ca(2+) transients. Steady-state intramembrane q(gamma) charge thus persists with modified kinetics that can be restored to its normally complex waveform by perchlorate, even in intact muscle fibres unable to release Ca(2+). It is thus unlikely that q(gamma) charge movement is a consequence of SR Ca(2+) release rather than changes in tubular membrane potential. PMID- 11897858 TI - Characterization of vasorelaxant responses to anandamide in the rat mesenteric arterial bed. AB - The endogenous cannabinoid anandamide has recently been identified as a vasorelaxant but the underlying mechanisms are controversial. The vasorelaxant responses to anandamide have now been examined in the rat mesenteric arterial bed. Anandamide caused potent vasorelaxations (pD(2) = 6.24 +/- 0.06; R(max) = 89.4 +/- 2.2 %) which were unaffected by inhibition of nitric oxide synthase with N(G)-nitro-L-arginine methyl ester (L-NAME; 300 microM). The responses were also predominantly endothelium independent and were unaffected by the cannabinoid CB(1) receptor antagonist SR141716A (1 microM), although at higher concentrations (3 and 10 microM) SR141716A was inhibitory. Both 1 mM ouabain (pD(2) = 5.90 +/- 0.07; R(max) = 50.4 +/- 6.5 %) and 100 microM 18alpha-glycyrrhetinic acid (pD(2) = 6.04 +/- 0.14; R(max) = 40.9 +/- 5.8 %) opposed anandamide-induced vasorelaxation. However, the gap junction inhibitors carbenoxolone (100 microM) and palmitoleic acid (50 microM) did not affect vasorelaxation to anandamide. Relaxation to anandamide was significantly attenuated by both capsaicin pretreatment to deplete the sensory nerves of neurotransmitters (pD(2) = 5.86 +/- 0.18; R(max) = 56.3 +/- 5.2 %) and the vanilloid antagonist ruthenium red (10 microM; pD(2) = 5.64 +/- 0.09; R(max) = 33.7 +/- 3.9 %). However, these inhibitory effects were prevented by the additional presence of L-NAME, when the relaxation to anandamide was unaffected (pD(2) = 6.19 +/- 0.07; R(max) = 81.9 +/- 2.8 %). The inhibitor of neuronal nitric oxide synthase, 7-nitroindazole, also prevented capsaicin from inhibiting the responses to anandamide. The results of this study point to anandamide acting via several mechanisms, which include the involvement of sensory nerves, but only in the presence of nitric oxide. PMID- 11897859 TI - Excitability changes in human corticospinal projections to forearm muscles during voluntary movement of ipsilateral foot. AB - Excitability of the H-reflex in the relaxed flexor carpi radialis (FCR) muscle was tested during voluntary oscillations of the ipsilateral foot at five evenly spaced delays during a 600 ms cycle. In some experiments the H-reflex was conditioned by transcranial magnetic stimulation (TMS). With the hand prone, the amplitude of the FCR H-reflex was modulated sinusoidally with the same period as the foot oscillation, the modulation peak occurring in coincidence with contraction of the foot plantar-flexor soleus and the trough during contraction of the extensor tibialis anterior. When the H-reflex was facilitated by TMS at short latency (conditioning-test interval: -2 to -3.5 ms), the modulation was larger than that occurring with an unconditioned reflex of comparable size. This suggests that both the peripheral and the corticospinal components of the facilitated response were modulated in parallel. When the H-reflex was tested 40 60 ms after conditioning, i.e. during the cortical "silent period" induced by TMS, no direct effect was produced on the reflex size but the foot-associated modulation was deeply depressed. These results suggest that the reflex modulation may depend on activity fluctuations in the cortical motor area innervating the forearm motoneurones. It is proposed that when the foot is rhythmically oscillated, along with the full activation of the foot cortical area a simultaneous lesser co-activation of the forearm area produces a subliminal cyclic modulation of cervical motoneurones excitability. Should the two limbs be moved together, the time course of this modulation would favour isodirectional movements of the prone hand and foot, indeed the preferential coupling observed when hand and foot are voluntarily oscillated. PMID- 11897860 TI - Central and peripheral mediation of human force sensation following eccentric or concentric contractions. AB - Fatigue was induced in the triceps brachii of the experimental arm by a regimen of either eccentric or concentric muscle actions. Estimates of force were assessed using a contralateral limb-matching procedure, in which target force levels (25 %, 50 % or 75 % of maximum) were defined by the unfatigued control arm. Maximum isometric force-generating capacity was reduced by 31 % immediately following eccentric contractions, and remained depressed at 24 (25 %) and 48 h (13 %) post-exercise. A less marked reduction (8.3 %) was observed immediately following concentric contractions. Those participants who performed prior eccentric contractions, consistently (at all force levels), and persistently (throughout the recovery period), overestimated the level of force applied by the experimental arm. In other words, they believed that they were generating more force than they actually achieved. When the forces applied by the experimental and the control arm, were each expressed as a proportion of the maximum force that could be attained at that time, the estimates matched extremely closely. This outcome is that which would be expected if the estimates of force were based on a sense of effort. Following eccentric exercise, the amplitude of the EMG activity recorded from the experimental arm was substantially greater than that recorded from the control arm. Cortically evoked potentials recorded from the triceps brachii (and extensor carpi radialis) of the experimental arm were also substantially larger than those elicited prior to exercise. The sense of effort was evidently not based upon a corollary of the central motor command. Rather, the relationship between the sense of effort and the motor command appears to have been altered as a result of the fatiguing eccentric contractions. It is proposed that the sense of effort is associated with activity in neural centres upstream of the motor cortex. PMID- 11897861 TI - Effects of prior contractions on muscle microvascular oxygen pressure at onset of subsequent contractions. AB - In humans, pulmonary oxygen uptake (.V(O2)) kinetics may be speeded by prior exercise in the heavy domain. This "speeding" arises potentially as the result of an increased muscle O(2) delivery (.Q(O2)) and/or a more rapid elevation of oxidative phosphorylation. We adapted phosphorescence quenching techniques to determine the.Q(O2)-to-O(2) utilization (.Q(O2)/.V(O2)) characteristics via microvascular O(2) pressure (P(O2,m)) measurements across sequential bouts of contractions in rat spinotrapezius muscle. Spinotrapezius muscles from female Sprague-Dawley rats (n = 6) were electrically stimulated (1 Hz twitch, 3-5 V) for two 3 min bouts (ST(1) and ST(2)) separated by 10 min rest. P(O2,m) responses were analysed using an exponential + time delay (TD) model. There was no significant difference in baseline and DeltaP(O2,m) between ST(1) and ST(2) (28.5 +/- 2.6 vs. 27.9 +/- 2.4 mmHg, and 13.9 +/- 1.8 vs. 14.1 +/- 1.3 mmHg, respectively). The TD was reduced significantly in the second contraction bout (ST(1), 12.2 +/- 1.9; ST(2), 5.7 +/- 2.2 s, P < 0.05), whereas the time constant of the exponential P(O2,m) decrease was unchanged (ST(1), 16.3 +/- 2.6; ST(2), 17.6 +/- 2.7 s, P > 0.1). The shortened TD found in ST(2) led to a reduced time to reach 63 % of the final response of ST(2) compared to ST(1) (ST(1), 28.3 +/- 3.0; ST(2), 20.2 +/- 1.8 s, P < 0.05). The speeding of the overall response in the absence of an elevated P(O2,m) baseline (which had it occurred would indicate an elevated.Q(O2)/.V(O2) or muscle blood flow suggests that some intracellular process(es) (e.g. more rapid increase in oxidative phosphorylation) may be responsible for the increased speed of P(O2,m) kinetics after prior contractions under these conditions. PMID- 11897862 TI - Areas of the brain concerned with ventilatory load compensation in awake man. AB - There is broad agreement that the awake human ventilatory response to a moderate inspiratory load consists of a prolongation of inspiratory time (T(I)) with a maintenance of tidal volume (V(T)) and end-tidal P(C)(O(2)) (P(ET,C)(O(2))), the response being severely blunted in sleep. There is no agreement on the mechanisms underlying this ventilatory response. Six naive healthy males (aged 39-44) were studied supine with their heads in a positron emission tomography (PET) scanner to allow relative regional cerebral blood flow (rCBF) to be measured with H(2)(15)O given intravenously. A linearised resistive load (24 cmH(2)O (l s(-1))( 1)) could be added to the inspiratory limb of a breathing valve inserted into a tightly fitting facemask; inspiratory flow was measured with a pneumotachograph. The load was applied, without alerting the subject, when the radioactivity first reached the head. Six scans were performed with and without the load, in each subject. Relative rCBF contrasts between the loaded and unloaded breathing states showed significant activations in inferior parietal cortex, prefrontal cortex, midbrain, basal ganglia and multiple cerebellar sites. No activations were found in the primary sensorimotor cortex. The findings suggest that there is a pattern of motor behavioural response to the uncomfortable sensation that inspiration is impeded. This results in a prolongation of T(I), the maintenance of V(T) and a reduction in the degree of discomfort, presumably because of the reduction of mean negative pressure in the airways. PMID- 11897863 TI - Stimulation of pulmonary vagal C-fibres by anandamide in anaesthetized rats: role of vanilloid type 1 receptors. AB - This study was carried out to determine the effect of intravenous injection of anandamide on pulmonary C-fibre afferents and the cardiorespiratory reflexes. In anaesthetized, spontaneously breathing rats, intravenous bolus injection of anandamide near the right atrium immediately elicited the pulmonary chemoreflex responses, characterized by apnoea, bradycardia and hypotension. After perineural treatment of both cervical vagi with capsaicin to block the conduction of C fibres, anandamide no longer evoked these reflex responses. In open-chest, and artificially ventilated rats, anandamide injection evoked an abrupt and intense discharge in vagal pulmonary C-fibres in a dose-dependent manner. After injection of the high dose, the fibre discharge generally started within 1 s, reached a peak in approximately 2 s, and returned to baseline within 7 s. The stimulation of C-fibres by anandamide was completely and reversibly blocked by pretreatment with capsazepine, a competitive antagonist of the vanilloid type 1 receptor. Anandamide (0.4 mg kg(-1)) stimulated approximately 93 % of pulmonary C-fibres that were activated by capsaicin at a much lower dose (0.6 microg kg(-1)); the response to anandamide showed similar intensity, but had slightly longer latency and duration than that to capsaicin. In conclusion, intravenous bolus injection of anandamide evokes a consistent and distinct stimulatory effect on pulmonary C fibre terminals, and this effect appears to be mediated through an activation of the vanilloid type 1 receptor. PMID- 11897864 TI - Developmental changes in cerebral autoregulatory capacity in the fetal sheep parietal cortex. AB - We validated laser Doppler flowmetry (LDF) for long-term monitoring and detection of acute changes of local cerebral blood flow (lCBF) in chronically instrumented fetal sheep. Using LDF, we estimated developmental changes of cerebral autoregulation. Single fibre laser probes (0.4 mm in diameter) were implanted in and surface probes were placed on the parietal cerebral cortex at 105 +/- 2 (n = 7) and 120 +/- 2 days gestational age (dGA, n = 7). Basal lCBF was monitored over 5 days followed by a hypercapnic challenge (fetal arterial partial pressure of CO(2), P(a,CO2): 83 +/- 3 mmHg) during which lCBF changes obtained by LDF were compared to those obtained with coloured microspheres (CMSs). Mean arterial blood pressure (MABP) was increased and decreased using phenylephrine and sodium nitroprusside at 110 +/- 2 and 128 +/- 2 dGA. Intracortical and cortical surface laser probes gave stable measurements over 5 days. The lCBF increase during hypercapnia obtained by LDF correlated well with flows obtained using CMS (r = 0.89, P < 0.01). The signals of intracortical and surface laser probes also correlated well (r = 0.91, P < 0.01). Gliosis of 0.35 +/- 0.06 mm around the tip of intracortical probes did not affect the measurements. The range of MABP over which cerebral autoregulation was observed increased from 20-48 mmHg at 110 dGA to 35 to > 95 mmHg at 128 dGA (P < 0.05). Since MABP increased from 33 to 54 mmHg over this period (P < 0.01), the range between the lower limit of cerebral autoregulation and the MABP increased from 13 mmHg at 110 dGA to 19 mmHg at 128 dGA (P < 0.01). LDF is a reliable tool to assess dynamic changes in cerebral perfusion continuously in fetal sheep. PMID- 11897867 TI - New feature: stem cells in the news. PMID- 11897865 TI - The effects of birth weight on basal cardiovascular function in pigs at 3 months of age. AB - In man, epidemiological studies have shown that low birth weight (BW) is associated with an increased risk of cardiovascular disease in later life. In this study, the long-term consequences of variations in natural BW on basal cardiovascular function were investigated in pigs at 3 months of postnatal age. Low (< 1.41 kg; n = 20) and high (> 1.52 kg; n = 20) BW Large White piglets were selected from a total of 12 litters for study at 3 months of age. Basal mean arterial pressure (MAP) and heart rate (HR) were recorded for approximately 30 min using standard recording equipment and basal arterial blood samples were taken for hormone analyses. Concentrations of angiotensin-converting enzyme (ACE) were also measured in kidney, lung and plasma. Basal MAP, but not HR, in 3-month old pigs was significantly inversely related to BW and positively related to the ratio of head length to BW. Postnatal growth rate of low BW pigs was slower than that of high BW pigs such that low BW piglets remained significantly smaller at 3 months of age. There were no differences in basal plasma adrenaline or cortisol concentrations between low and high BW pigs. However, basal plasma noradrenaline concentrations were significantly elevated in low BW compared to high BW pigs. Renal and pulmonary ACE levels were significantly reduced in low BW compared to high BW pigs. These data show that basal MAP in 3-month-old pigs is negatively associated with BW and positively correlated to disproportionate size at birth. This effect was associated with an increase in basal plasma noradrenaline concentrations. PMID- 11897868 TI - Gene repair and transposon-mediated gene therapy. AB - The main strategy of gene therapy has traditionally been focused on gene augmentation. This approach typically involves the introduction of an expression system designed to express a specific protein in the transfected cell. Both the basic and clinical sciences have generated enough information to suggest that gene therapy would eventually alter the fundamental practice of modern medicine. However, despite progress in the field, widespread clinical applications and success have not been achieved. The myriad deficiencies associated with gene augmentation have resulted in the development of alternative approaches to treat inherited and acquired genetic disorders. One, derived primarily from the pioneering work of homologous recombination, is gene repair. Simply stated, the process involves targeting the mutation in situ for gene correction and a return to normal gene function. Site-specific genetic repair has many advantages over augmentation although it too is associated with significant limitations. This review outlines the advantages and disadvantages of gene correction. In particular, we discuss technologies based on chimeric RNA/DNA oligonucleotides, single-stranded and triplex-forming oligonucleotides, and small fragment homologous replacement. While each of these approaches is different, they all share a number of common characteristics, including the need for efficient delivery of nucleic acids to the nucleus. In addition, we review the potential application of a novel and exciting nonviral gene augmentation strategy--the Sleeping Beauty transposon system. PMID- 11897869 TI - Ligand/receptor signaling threshold (LIST) model accounts for gp130-mediated embryonic stem cell self-renewal responses to LIF and HIL-6. AB - We previously demonstrated that embryonic stem (ES) cell self-renewal required sustained signaling by leukemia inhibitory factor (LIF) in a concentration dependent manner, allowing us to hypothesize that thresholds in ligand-receptor signaling modulate stem cell differentiation control. To test this hypothesis, we have experimentally and computationally compared the abilities of two gp130 signaling cytokines (LIF and Hyper-interleukin-6 [HIL-6]) to sustain ES cell self renewal. Quantitative measurements of ES cell phenotypic markers (stage-specific embryonic antigen-1 and E-cadherin), functional assays (alkaline phosphatase activity and embryoid body formation efficiency), and transcription factor (Oct 4) expression over a range of LIF and HIL-6 concentrations demonstrated a superior ability of LIF to maintain ES cell pluripotentiality at higher concentrations (> or =500 pM). Additionally, we observed distinct qualitative differences in the ES cell self-renewal dose response profiles between the two cytokines. A computational model permitted calculation of the number of signaling complexes as a function of receptor expression, ligand concentration, and ligand/receptor-binding properties, generating predictions for the degree of self renewal as a function of cytokine concentration by comparison of these calculated complex numbers to experimentally determined threshold cytokine concentrations. Model predictions, consistent with experimental data, indicated that differences in the potencies of these two cytokines were based primarily on differences in receptor-binding stoichiometries and properties. These results support a ligand/receptor signaling threshold model of ES cell fate modulation through appropriate types and levels of cytokine stimulation. Insights from these results may be more generally applicable to tissue-specific stem cells and could aid in the development of stem cell-based technologies. PMID- 11897870 TI - Analysis of different promoter systems for efficient transgene expression in mouse embryonic stem cell lines. AB - Mouse embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo and have the developmental capacity to generate all cell types of the body. Combined with efficient genetic manipulation and in vitro differentiation procedures, ES cells are a useful system for the molecular analysis of developmental pathways. We analyzed and compared the transcriptional activities of a cellular polypeptide chain elongation factor 1 alpha (EF), a cellular-virus hybrid (cytomegalo-virus [CMV] immediate early enhancer fused to chicken beta-actin [CBA]), and a viral CMV promoter system in two ES cell lines. When transiently transfected, the EF and CBA promoters robustly drove reporter gene expression, while the CMV promoter was inactive. We also demonstrated that the EF and CBA promoters effectively drove gene expression in different stages of cell development: naive ES cells, embryoid bodies (EBs), and neuronal precursor cells. In contrast, the CMV promoter did not have transcriptional activity in either ES cells or EB but had significant activity once ES cells differentiated into neuronal precursors. Our data show that individual promoters have different abilities to express reporter gene expression in the ES and other cell types tested. PMID- 11897871 TI - In vitro differentiation of embryonic stem cells into hepatocyte-like cells identified by cellular uptake of indocyanine green. AB - BACKGROUND AND AIMS: Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro. We have recently found the emergence of cell clusters that show the cellular uptake of indocyanine green (ICG) in the culture of differentiated ES cells. ICG is clinically used as a test substance to evaluate liver function because it is eliminated exclusively by hepatocytes. The aim of the present study was to investigate the hepatic characteristics of ICG-stained cells. METHODS: Embryoid bodies (EBs), formed by a 5-day hanging drop culture of ES cells, were allowed to outgrow in the placed culture. Gene expression of hepatocyte markers was analyzed by reverse transcriptase-polymerase chain reaction, and albumin production was examined immunohistochemically. Morphology and cellular components were investigated by electron microscopy. ICG-stained cells were further transplanted into the portal vein of mice. RESULTS: ICG-stained cells appeared around 14 days of the EB culture and formed distinct three-dimensional structures. They were immunoreactive to albumin and expressed mRNAs such as albumin, alpha-fetoprotein, transthyretin, hepatocyte nuclear factor 3 beta, alpha-1-antitrypsin, tryptophan 2,3-dioxygenase, urea cycle enzyme, gluconeogenic enzyme, and liver-specific organic anion transporter-1. An ultrastructural analysis revealed a well developed system of organelles such as mitochondria, lysosomes, Golgi apparatus, and rough and smooth endoplasmic reticulum. The transplantation of ICG-positive cells into the portal vein resulted in the incorporation into mice livers, where they were morphologically indistinguishable from neighboring hepatocytes. CONCLUSIONS: ES cell-derived ICG-positive cells possess characteristics of hepatocytes, and ICG-staining is a useful marker to identify differentiated hepatocytes from EBs in vitro. PMID- 11897872 TI - Comparison of bone marrow cells harvested from various bones of cynomolgus monkeys at various ages by perfusion or aspiration methods: a preclinical study for human BMT. AB - Using cynomolgus monkeys, we have previously established a new method for harvesting bone marrow cells (BMCs) with minimal contamination of the BMCs with T cells from the peripheral blood. We originally conducted this new "perfusion method" in the long bones (the humerus, femur, and tibia) of cynomolgus monkeys. Here, we apply the perfusion method to obtain BMCs from the ilium of cynomolgus monkeys, since BMCs are usually collected from the ilium by the conventional aspiration method in humans. The perfusion method consists of two approaches: transverse iliac perfusion and longitudinal iliac perfusion. BMCs harvested by the perfusion method from the long bones and ilium were compared with those collected from the ilium by the aspiration method. The contamination of BMCs with peripheral blood, determined by the frequencies of CD4+ and CD8+ T cells, was significantly lower in BMCs obtained from the ilium or long bones by the perfusion method (CD4+ plus CD8+ T cells <4%) than in those obtained by the iliac aspiration method (CD4+ plus CD8+ T cells >20%). However, the numbers of immature myeloid cells, such as myeloblasts, promyelocytes, myelocytes, and metamyelocytes, were higher in BMCs obtained by the iliac perfusion method than in those obtained by the iliac aspiration method. The assays for in vitro colony forming unit in culture revealed that progenitor activity was significantly higher in BMCs obtained by the perfusion method than in those obtained by the aspiration method. These findings suggest that the contamination of BMCs with peripheral blood is much less when using the perfusion method than when using the aspiration method. To determine the best site for harvesting BMCs by the perfusion method, age-dependent changes in BMCs harvested by the perfusion method from the long bones and ilium were examined. The numbers of BMCs varied in the long bones (humerus > femur > tibia) and showed age-dependent decreases, whereas they remained similar in the ilium of cynomolgus monkeys from 3 years to 6 years of age. However, in cynomolgus monkeys, BMC harvesting by the perfusion method from the ilium (but not from the long bones) is found to involve the risk of fat emboli, particularly when the BMCs are quickly perfused under high pressure. These findings suggest, even in humans, that the perfusion method is better than the aspiration method, and that the best site for collection of BMCs is the humerus. PMID- 11897873 TI - Long-term survival and glial differentiation of the brain-derived precursor cell line RN33B after subretinal transplantation to adult normal rats. AB - The potential use of in vitro-expanded precursor cells or cell lines in repair includes transplantation of such cells for cell replacement purposes and the activation of host cells to provide "self-repair." Recently, we have reported that cells from the brain-derived cell line RN33B (derived from the embryonic rat medullary raphe and immortalized through retroviral transduction of the temperature-sensitive mutant of the simian virus 40 ([SV40] large T-antigen) survive for at least 4 weeks, integrate, and differentiate after subretinal grafting to normal adult rats. Here, we demonstrate that grafts of these cells survive for at least 4 months after subretinal transplantation to adult, normal immunosuppressed rats. Implanted cells integrate into the retinal pigment epithelium and the inner retinal layers, and the anterior part of the optic nerve. In addition, the RN33B cells migrate within the retina, occupying the whole retina from one eccentricity to the other. A large fraction of the grafted cells differentiate into glial cells, as shown by double labeling of the reporter genes LacZ or green fluorescent protein, and several glial markers, including oligodendrocytes. However, the cells did not differentiate into retinal neurons, judging from their lack of expression of retinal neuronal phenotypic markers. A significant number of the implanted cells in the host retina were in a proliferative stage, judging from proliferative cell nuclear antigen and SV40 large T-antigen immunohistochemistry. To conclude, the cells survived, integrated, and migrated over long distances within the host. Therefore, our results may be advantageous for future design of therapeutic strategies, since such cells may have the potential of being a source of, for example, growth factor delivery in experimental models of retinal degeneration. PMID- 11897874 TI - Human CD34+ hematopoietic stem/progenitor cells express high levels of FLIP and are resistant to Fas-mediated apoptosis. AB - We sought to determine whether lympho-hematopoietic stem-progenitor cells (HSC) from human placental/umbilical cord blood (CB) or adult mobilized blood (PBSC) are sensitive to Fas-induced apoptosis. Human CD34+ cells from CB or PBSC were cultured in serum-free medium, with or without hematopoietic growth factors (FKT: FLT-3 ligand [FL], KIT ligand [KL], and thrombopoietin [TPO]), and with or without soluble Fas ligand (sFasL) or agonistic anti-Fas antibody. After 5-48 hours of culture, cells were assessed for viability and stained with Annexin V and 7-Aminoactinomycin D for apoptosis analysis by fluorescence-activated cell sorting. Cultured cells were also assessed by in vitro hematopoietic colony forming cell (CFC) and in vivo nonobese diabetic/severe combined immunodeficient mouse engraftment potential (SEP) assays. Levels of Fas, FLICE inhibitory protein (FLIP), and Caspase 8 mRNA in CD34+ cells were determined by real-time quantitative polymerase chain reaction. Expression of FLIP was confirmed by Western blotting. No decrease in viability, CFC, or SEP was observed in CB or PBSC CD34+ cells cultured in the presence of sFasL or agonistic anti-Fas antibody. Human CB and mobilized PBSC CD34+ cells expressed high levels of FLIP, low ratios of Caspase 8:FLIP, and low levels of Fas. Thus, human CB and PBSC CD34+ HSC were resistant to Fas pathway agonists. High-level expression of FLIP likely provides one level of protection of CD34+ cells from Fas-mediated apoptosis. PMID- 11897875 TI - Ex vivo expansion of stem cells from umbilical cord blood: expression of cell adhesion molecules. AB - Expansion of stem cells from cord blood has been demonstrated to increase the numbers of CD34+ cells, CD34+ subsets, long-term culture-initiating cells, and severe combined immunodeficient mouse, repopulating cells. However, reports suggest that the ex vivo expanded population behaves differently than freshly isolated cells and shows a delayed or diminished engraftment. In this study, we investigated the effects of the cytokines flt3 ligand, stem cell factor, and thrombopoietin on expansion of CD34+ and CD34+/CD38- cells. In addition, we studied the expression of adhesion molecules, very late activation antigen-4 (VLA 4) and leukocyte function antigen-1 (LFA-1), on CD34+ cells from cord blood by flow cytometry. We also looked at the expression of an adhesion receptor, namely, vascular cell adhesion molecule-1 (VCAM-1) on bone marrow stromal cells by Western blot analysis after exposure to low dose gamma irradiation. After culturing for 7 days, increases in the absolute numbers of CD34+, CD34+/CD38-, CD34+/VLA-4+, and CD34+/LFA-1+ cells were 5.67 +/- 2.91 (mean +/- standard deviation) fold, 7.21 +/- 4.38 fold, 99.56 +/- 101.5 fold, and 101.39 +/- 83.25 fold, respectively. There was a transient upregulation in the expression levels of VCAM-1 on stromal cells, which peaked at 4 hours. Though there was an increase in the absolute numbers of CD34+ cells expressing the adhesion molecules, the expression levels (antigen density) of the adhesion molecules on the CD34+ cells remained unaffected. PMID- 11897876 TI - The molecular perspective: restriction endonucleases. PMID- 11897877 TI - Nutrition and athletic performance. AB - This supplement is based on presentations made at the "First International Congress on Nutrition and Athletic Performance" held at the Shaw Conference Centre, Edmonton, Alberta, Canada, August 8-11, 2001. This conference was stimulated by an increasing awareness of the potential benefits and risks of diet modification and supplementation for elite athletic performance. This supplement summarizes recent findings and commentary on new research directions in nutrition and athletic performance by 22 invited plenary speakers from around the world. PMID- 11897878 TI - Update on thiol status and supplements in physical exercise. AB - Strenuous physical exercise represents a condition that is often associated with increased production of reactive oxygen species in various tissues. One of the most reliable indices of exercise-induced oxidant production is tissue glutathione oxidation. In humans, exercise-induced blood glutathione oxidation is rapid and subject to control by antioxidant supplementation. The objective of this brief review is to provide an update of our current understanding of cellular thiols and thiol antioxidants. Cellular thiols are critically important in maintaining the cellular antioxidant defense network. In addition, thiols play a key role in regulating redox-sensitive signal transduction process. Lipoic acid is a highly promising thiol antioxidant supplement. Recent studies have clarified that while higher levels of oxidants may indeed inflict oxidative damage, oxidants are not necessarily deleterious. Under certain conditions oxidants may function as cellular messengers that regulate a multitude of signal transduction pathways. In light of this, the significance of oxidants in various aspects of biology needs to be revisited. PMID- 11897879 TI - Magnesium, zinc, and chromium nutrition and athletic performance. AB - Magnesium, zinc and chromium are mineral elements required in modest amounts for health and optimal performance. Accumulating evidence supports the hypothesis that magnesium and zinc play significant roles in promoting strength and cardiorespiratory function in healthy persons and athletes. Differences in study designs, however, limit conclusions about recommendations for intakes of magnesium and zinc needed for optimal performance and function. The role of chromium in supporting performance is not well established. There is a compelling need to confirm and extend findings of beneficial effects of magnesium and zinc function and performance of humans. Suggestions for an experimental model and specific topics for research to advance knowledge of mineral nutrition to promote attainment of genetic potential for optimal performance are provided. PMID- 11897880 TI - Nutritional strategies to minimise exercise-induced immunosuppression in athletes. AB - Strenuous prolonged exertion and heavy training are associated with depressed immune function. Furthermore, improper nutrition can compound the negative influence of heavy exertion on immunocompetence. Dietary deficiencies of protein and specific micronutrients have long been associated with immune dysfunction. An adequate intake of iron, zinc, and vitamins A, E, B6 and B12 is particularly important but excess intakes can also impair immune function. Immune system impairment has also been associated with excess intake of fat. To maintain immune function, athletes should eat a well balanced diet sufficient to meet their energy requirements. An athlete exercising in a carbohydrate-depleted state experiences larger increases in circulating stress hormones and a greater perturbation of several immune function indices. Conversely, consuming carbohydrate during exercise attenuates rises in stress hormones such as cortisol and appears to limit the degree of exercise-induced immunosuppression, at least for non-fatiguing bouts of exercise. Strong evidence that high doses of antioxidant vitamins, glutamine supplementation or echinacea extracts can prevent exercise-induced immunosuppression is lacking. PMID- 11897882 TI - Exercise immunology: nutritional countermeasures. AB - In contrast to moderate physical activity, prolonged and intensive exertion causes numerous changes in immunity that reflect physiologic stress and suppression, and an increased risk of upper respiratory tract infection. Enzymes in immune cells require the presence of micronutrients, leading to attempts by investigators to alter changes in immunity following heavy exertion through use of nutritional supplements, primarily zinc, dietary fat, vitamin C and other antioxidants, glutamine, and carbohydrate. Except for carbohydrate supplementation, none of these nutrients has emerged as an effective countermeasure to exercise-induced immunosuppression. Data from several studies of endurance athletes suggest that carbohydrate compared to placebo ingestion is associated with an attenuated cortisol, growth hormone, and epinephrine response to heavy exertion, fewer perturbations in blood immune cell counts, lower granulocyte and monocyte phagocytosis and oxidative burst activity, and a diminished pro- and anti-inflammatory cytokine response. Overall, the hormonal and immune responses to carbohydrate compared to placebo ingestion during intensive exercise suggest that physiologic stress and inflammation are diminished, although clinical significance awaits further research. PMID- 11897881 TI - Do we have unrealistic expectations of the potential of immuno-nutrition? AB - Heavy sports training schedules and competition is often associated with immuno suppression, and so there is a theoretical justification for providing athletes with nutrients that display immuno-regulatory properties. Among such immuno nutrients, considerable attention has been paid in recent years to two amino acids, arginine (ARG) and glutamine (GLN). ARG and GLN availability regulate the function of T lymphocytes, macrophages and polymorphonuclear cells. ARG acts through nitric oxide and polyamine synthesis. The mechanism of action of GLN in immune cells remains unclear. Experience in clinical nutrition suggests that an ARG-enriched diet may limit infectious morbidity in critically ill patients. Data concerning oral/enteral GLN supplementation are more controversial. There have been few trials of supplementation in sports medicine, but results are promising, justifying further studies in which dosages and administration schedules should be taken into account. PMID- 11897883 TI - Diet and training in the week before competition. AB - For many years athletes have used carbohydrate loading to enhance endurance performance. This practice has been based on findings demonstrating that 1) exercise induced depletion of the muscle glycogen stores followed by the intake of a carbohydrate rich diet, resulted in muscle glycogen stores above normal values and 2) that the pre-exercise muscle glycogen content was the main determinant of the capacity to perform strenuous exercise to exhaustion. Lately it has been speculated whether a period of a high fat diet, followed by carbohydrate loading to restore or increase muscle glycogen levels above normal, would be of further advantage for exercise performance. From the discussed data it emerges, however, that varying periods of fat adaptation followed by a carbohydrate rich diet prior to exercise is of no benefit for exercise performance. Despite an increased fat oxidation and a concomitant decrease in carbohydrate oxidation during submaximal exercise, no benefit in a subsequent time trial appeared. Data suggest that this dietary regimen impairs the ability to utilise carbohydrates. PMID- 11897884 TI - Pre-exercise nutritional strategies: effects on metabolism and performance. AB - The goals of pre-exercise nutritional strategies are to optimise the availability of carbohydrate (CHO) and fluid. Ingestion of CHO 3-4 hr prior to exercise can increase liver and muscle glycogen stores and has been associated with enhanced endurance exercise performance. The metabolic effects of CHO ingestion persist for at least 6 hr. Although an increase in plasma insulin following CHO ingestion in the hour prior to exercise inhibits lipolysis and liver glucose output, and can lead to transient hypoglycemia during subsequent exercise, there is no convincing evidence that this is always associated with impaired exercise performance. Having said that, individual experience should inform individual practice. Interventions to increase plasma FFA availability prior to exercise have been shown to reduce CHO utilisation during exercise, but do not appear to have major ergogenic benefits. It is more difficult to hyperhydrate prior to exercise and although there has been interest in glycerol ingestion, to date research results have been equivocal. At the very least, athletes should ensure euhydration prior to exercise. PMID- 11897885 TI - Food and fluid intake during exercise. AB - The intake of fluid and CHO offers benefits to the performance of a number of sports events and exercise activities. The effects of dehydration on performance are now well known, with the penalties ranging from subtle, but often important, decrements in performance at low levels of fluid deficit to the severe health risks associated with substantial fluid losses during exercise in the heat. Although evidence of the beneficial effects of CHO intake during exercise have existed for over 70 years, sports scientists are still to discover all the situations in which benefits occur and to explain the mechanisms involved. Optimal strategies for CHO and fluid intake during exercise are yet to be fine tuned, and ultimately will be determined by practical issues such as the opportunity to eat or drink during an event, and gastrointestinal comfort. PMID- 11897886 TI - Creatine supplementation: exploring the role of the creatine kinase/phosphocreatine system in human muscle. AB - The effect of oral creatine supplementation on high-intensity exercise performance has been extensively studied over the past ten years and its ergogenic potential in young healthy subjects is now well documented. Recently, research has shifted from performance evaluation towards elucidating the mechanisms underlying enhanced muscle functional capacity after creatine supplementation. In this review, we attempt to summarise recent advances in the understanding of potential mechanisms of action of creatine supplementation at the level of skeletal muscle cells. By increasing intracellular creatine content, oral creatine ingestion conceivably stimulates operation of the creatine kinase (CK)/phosphocreatine (PCr) system, which in turn facilitates muscle relaxation. Furthermore, evidence is accumulating to suggest that creatine supplementation can beneficially impact on muscle protein and glycogen synthesis. Thus, muscle hypertrophy and glycogen supercompensation are candidate factors to explain the ergogenic potential of creatine ingestion. Additional issues discussed in this review are the fibre-type specificity of muscle creatine metabolism, the identification of responders versus non-responders to creatine intake, and the scientific background concerning potential side effects of creatine supplementation. PMID- 11897887 TI - Caffeine, coffee and ephedrine: impact on exercise performance and metabolism. AB - This paper addresses areas where there is controversy regarding caffeine as an ergogenic aid and also identifies topics that have not been adequately addressed. It is clear that caffeine, in moderate amounts, can be used orally as an ergogenic aid in aerobic activity lasting for more than 1 min. It increases endurance and speed, but not maximal VO2 and related parameters. While there are fewer well-controlled studies for resistance exercise, the literature would suggest similar improvements: increased endurance at submaximal tension and power generated in repeated contractions and no change in maximal ability to produce force. It is likely that theophylline (a related methylxanthine) has similar actions and it has been suggested that the combination of caffeine and sympathomimetics may be a more potent erogenic aid. The voids in our understanding of caffeine include the dose (what amount is optimal, what vehicle is used to deliver the drug as well as method, pattern, and mode of administration), the potential side effects (particularly in competitive settings), health implications (insulin resistance and if combined with ephedrine, cardiovascular risks) and mechanisms of action. It appears unlikely that increased fat oxidation and glycogen sparing is the prime ergogenic mechanism. PMID- 11897888 TI - Sport nutritional supplements: quality and doping controls. AB - Nutritional supplements are part of the diet of many athletes. With the exception of caffeine and ephedrine alkaloids, most of these products do not contain substances that are prohibited to competing sportsmen. In recent years, androgens, pro-hormones such as DHEA, androstenedione, androstenediol and 19 norsteroids became available for oral self-administration in many countries and on the Internet. Their claimed actions, efficiency or potency, and the possible adverse effects have not been thoroughly investigated by controlled clinical studies. Some products were shown to contain prohibited substances such as ephedrine, caffeine, or steroids, that were not listed on the label. Urine samples collected after the administration of these supplements can test positive. The administration of natural steroids such as testosterone and its precursors cannot be proven by the sole identification of the substances in the urine. The approach to detection is based upon the deviation of selected parameters of the metabolic profiles from the range of values normally found in humans. The individual's norm is also studied to exclude the few cases of systematic and natural excretion of extreme values. The combination of the GC/MS and the GC/C/IRMS offers a powerful tool to discriminate between the natural and synthetic origin of the urinary steroids. PMID- 11897890 TI - Protein nutrition and resistance exercise. AB - Strength conditioning will result in an increase in muscle size and this increase in size is largely the result of increased contractile proteins. The mechanisms by which the mechanical events stimulate an increase in RNA synthesis and subsequent protein synthesis are not well understood. Lifting weight requires that a muscle shorten as it produces force (concentric contraction). Lowering the weight forces the muscle to lengthen as it produces force (eccentric contraction). Eccentric contractions produce ultrastructural damage that may stimulate increased muscle protein turnover and a cascade of metabolic events which is similar to an acute phase response and includes complement activation, mobilization of neutrophils, increased circulating and skeletal muscle interleukin-1 and macrophage accumulation. While endurance exercise increases the oxidation of essential amino acids and increases the requirement for dietary protein, resistance exercise results in a decrease in nitrogen excretion, lowering dietary protein needs. Research has indicated that increased dietary protein intake (up to 1.6 g protein x kg(-1) x d(-1)) may enhance the hypertrophic response to resistance exercise. It has also been demonstrated that in very old men and women the use of a protein-calorie supplement was associated with greater strength and muscle mass gains than did the use of placebo. PMID- 11897891 TI - Protein intake and bone growth. AB - Among osteotrophic nutrients, proteins play an important role in bone development, thereby influencing peak bone mass. Consequently, protein malnutrition during development can increase the risk of osteoporosis and of fragility fracture later in life. Both animal and human studies indicate that low protein intake can be detrimental for both the acquisition of bone mass during growth and its conservation during adulthood. Low protein intake impairs both the production and action of IGF-I (Insulin-like growth factor-I). IGF-I is an essential factor for bone longitudinal growth, as it stimulates proliferation and differentiation of chondrocytes in the epiphyseal plate, and also for bone formation. It can be considered as a key factor in the adjustments of calcium phosphate metabolism required for normal skeletal development and bone mineralization during growth. In healthy children and adolescents, a positive association between the amount of ingested proteins and bone mass gain was observed in both sexes at the level of the lumbar spine, the proximal femur and the midfemoral shaft. This association appears to be particularly significant in prepubertal children. This suggests that, like for the bone response to either the intake of calcium or weight-bearing exercise, the skeleton would be particularly responsive to the protein intake during the years preceding the onset of pubertal maturation. PMID- 11897889 TI - Protein and amino acid requirements of adults: current controversies. AB - Protein intakes vary widely but costs and benefits of such variation is a long standing unresolved issue. The wide range of reported values for the minimum protein intake for N equilibrium in adults, i.e. 0.39 to 1.09 g/kg is best explained by an Adaptive Metabolic Demands model in which metabolic demands include amino acid oxidation at a rate varying with habitual protein intake and which changes slowly with dietary change. Thus within the reported data the true minimum requirement intake, the lowest values in the range at intakes approaching the Obligatory Nitrogen Loss, allows only fully adapted subjects to achieve N equilibrium. The higher values reflect incomplete adaptation. (13)C-1 leucine tracer balance studies of this model show (a) a fall with age in apparent protein requirements, (b) better than predicted efficiency of wheat protein utilization, and (c) controversially, lower lysine requirements than other workers, consistent with new evidence of de novo synthesis of lysine from urea salvaged by large bowel microflora. The main implication of the requirements model for athletes on high protein diets is increased exercise induced amino acid oxidation and risk of loss of body N when such high intakes are not maintained. PMID- 11897892 TI - The concept of energy homeostasis for optimal health during training. AB - From all nutritional variables optimal energy supply is considered as most vital for human performance. It is postulated that lack of energy homeostasis is the basic problem in the development of overtraining. Most if not all clinical symptoms are directly or indirectly related to the physiological mechanisms of energy homeostasis. The rapidly increasing knowledge in the field of body weight control with several new regulatory neuro-peptides such as leptin, will give new opportunities to tackle this unbalance between training load and energy availability. The central role of leptin and insulin as adiposity signals has focussed attention on the anti-obesity aspects of leptin. However as member of the cytokine family, leptin is also closely linked to the immune and reproductive system. New data indicates clearly the dual function of leptin at both ends at the energy balance; starvation vs. overfeeding. It links also nutrition to the reproductive system. Lack of available energy has a much greater impact on leptin levels than exercise stress. It is suggested that application of the rapidly increasing knowledge in the obesity field will benefit the research on the mechanisms involved in the derailment of the delicate balance between training load and energy homeostasis in athletes. PMID- 11897893 TI - Physical health of the female athlete: observations, effects, and causes of reproductive disorders. AB - This review begins by summarizing the state of knowledge about menstrual disorders in athletes at the turn of the 21st Century. It then highlights the most important developments of outstanding interest that have been reported in the 18 months since then. New observations of the characteristics of these disorders are followed by new reports of clinical consequences and recommendations for treatment, and discoveries about their physiological mechanism. In general, evidence is continuing to accumulate that exercise has no suppressive effect on the reproductive system beyond the impact of its energy cost on energy availability. These results encourage the hope that athletic women may be able to prevent or reverse menstrual disorders by dietary supplementation without any moderation of their exercise regimen. PMID- 11897894 TI - Nutritional considerations for the child athlete. AB - While nutritional issues are similar for all athletes irrespective of age, children have several physiological characteristics that distinguish them from adults and require specific nutritional considerations. These age- or maturation related differences include: a greater need for protein intake, to support growth; a greater need for calcium intake, to support bone accretion; a higher energy cost of activities that include walking and running; lower losses of sodium and chloride in sweat; and a greater thermoregulatory strain at any given level of hypohydration. This review will focus on three areas: (a) a higher metabolic cost of locomotion, its causes, and possible relevance to the calculation of daily energy requirements for young athletes; (b) the effect of carbohydrate (glucose, glucose plus fructose, or glucose plus sucrose) ingestion on children's aerobic performance, substrate utilization, and immune responses; and (c) involuntary dehydration during exercise in hot climate and the means for its prevention. PMID- 11897895 TI - The adverse effects of elite competition on health and well-being. AB - It is often assumed that participation in sport will produce only an array of health benefits. The adverse consequences of sport participation, particularly at the elite level, are rarely explored. Evidence continues to accumulate of a variety of unfortunate consequences that may accompany elite sport participation. Sport involvement may exacerbate pre-existing health problems, cause injury or even death. The sport environment may be hazardous in a variety of physical, emotional, and social ways. The common training and competition practices of certain sport cultures may themselves be harmful. Athletes may sacrifice health, home, education and normal social development in the pursuit of sport "success." Sport medicine professionals and sport scientists have particular opportunities and responsibilities to act as an athlete's advocate--and to protect their health and well being. PMID- 11897896 TI - Energy needs of athletes. AB - Each athlete has unique energy requirements, which underpin their ability to meet total nutritional goals. For everyday dietary planning and evaluation, energy requirements can be predicted via estimations of RMR and activity levels. Research methods such as indirect calorimetry and DLW allow energy requirements to be measured, and may be useful to confirm situations in which an athlete has a true energy balance anomaly. There is some evidence that individual athletes may have reduced energy requirements, although this occurs less frequently than is reported. Most self-reports of food intake substantially under-estimate energy intake, due to under-reporting or under-eating during the period of record keeping. Many athletes are over-focused on reducing body mass and body fat below levels that are consistent with long-term health and performance. Restrained eating can cause significant detrimental outcomes to body function. Leptin may be involved in modulating or mediating some of these changes. Athletes should use their energy budget to choose foods that provide macronutrient and micronutrient needs for optimal health and performance. Practical advice may help athletes to achieve energy intake challenges. PMID- 11897897 TI - Effects of amino acid intake on anabolic processes. AB - In the resting state muscle protein breakdown exceeds the rate of muscle protein synthesis, meaning that the balance between synthesis and breakdown is negative. Resistance exercise improves the net balance by stimulating muscle protein synthesis, but nutrient intake is required for synthesis to exceed breakdown (i.e., an anabolic response). Exercise and exogenous amino acids have an additive effect on muscle protein synthesis. There is a time-course of the response to a steady-state change in amino acid concentration. The signal for stimulation of muscle protein synthesis appears to be the extracellular concentrations of one or more of the essential amino acids (EAAs). Further, the rate, and direction, of change in extracellular concentrations (rather than the static concentration, per se) may be the important. Ingestion of non-essential AAs is not needed to stimulate muscle protein synthesis. Carbohydrate has, at most, a modest effect to enhance the response to amino acid ingestion after exercise. Finally, a mixture of EAAs + CHO more effectively stimulates muscle protein synthesis when taken before as opposed to after exercise. PMID- 11897898 TI - Restoration of fluid and electrolyte balance after exercise. AB - Post-exercise restoration of fluid balance after sweat-induced hypohydration avoids the detrimental effects of a body water deficit on physiological function and subsequent exercise performance. For effective restoration of fluid balance, the consumption of a volume of fluid in excess of the sweat loss and replacement of electrolyte, particularly sodium, losses are essential. Intravenous fluid replacement after exercise has been investigated to a lesser extent and its role for fluid replacement in the dehydrated but otherwise well athlete remains equivocal. PMID- 11897899 TI - Dietary strategies to promote glycogen synthesis after exercise. AB - Muscle glycogen is an essential fuel for prolonged intense exercise, and therefore it is important that the glycogen stores be copious for competition and strenuous training regimens. While early research focused on means of increasing the muscle glycogen stores in preparation for competition and its day-to-day replenishment, recent research has focused on the most effective means of promoting its replenishment during the early hours of recovery. It has been observed that muscle glycogen synthesis is twice as rapid if carbohydrate is consumed immediately after exercise as opposed to waiting several hours, and that a rapid rate of synthesis can be maintained if carbohydrate is consumed on a regular basis. For example, supplementing at 30-min intervals at a rate of 1.2 to 1.5 g CHO x kg(-1) body wt x h(-1) appears to maximize synthesis for a period of 4- to 5-h post exercise. If a lighter carbohydrate supplement is desired, however, glycogen synthesis can be enhanced with the addition of protein and certain amino acids. Furthermore, the combination of carbohydrate and protein has the added benefit of stimulating amino acid transport, protein synthesis and muscle tissue repair. Research suggests that aerobic performance following recovery is related to the degree of muscle glycogen replenishment. PMID- 11897901 TI - Functional genomics in reproductive medicine. AB - The British Fertility Society organised a workshop on Functional Genomics in Reproductive Medicine at the University of Birmingham on 13-14 September 2001. The primary aim was to inform delegates about the power of the technology that has been made available after completion of the sequencing of the human genome, and to stimulate debate about using functional genomics to address both clinical and scientific questions in reproductive medicine. Three specific areas were addressed: proteomics, gene expression and bioinformatics. Although the sophistication and plethora of techniques available were obvious, major limitations in the technology were also discussed. The future promises to be very challenging indeed. PMID- 11897902 TI - Lessons from a recent adoption study to identify some of the service needs of, and issues for, donor offspring wanting to know about their donors. AB - This paper draws on some of the major findings of a recent large-scale study of over 400 adult adopted people, who either searched for origins information or were sought out by birth relatives, to identify the potential profile of donor offspring seeking origins information. It is predicted that more women than men will search, that people who search will be in their twenties or older, and that the age at which searching begins may be delayed by the effects of the social stigma attached to gamete donation and by the greater likelihood of accidental disclosure in adulthood resulting from the higher incidence of secrecy about donor assisted conception. Two of the single triggers for adopted people to begin searching (as opposed to multiple triggers) - becoming a parent and the death of adoptive parents - may also be among the triggers for donor offspring to begin searching. The search may be complicated further when undertaken after accidental disclosure. Finally, it is argued that some donor offspring will experience a normative urge for identity completion and seeking relationships similar to that experienced by adopted people. This urge may stem from the fact that some donor offspring attach an identity to their donor that extends beyond needing factual details about their physical characteristics (though not necessarily a desire to establish a relationship). Some donor offspring are likely to encounter a desire for face-to-face contact, regardless of whether a face-to-face meeting was the original intention. The need for services to help donor offspring, donors, family members and others affected by the situation is identified. PMID- 11897903 TI - The extended role of the nurse: practical realities. AB - This article provides an overview of the development of the extended role of fertility nurses. The key factors discussed are the historical development of the nurses' role in the Oxford Fertility Unit at John Radcliffe Hospital and the relevant issues that influence this process and how this is reflected in other units nationally. Each practitioner is accountable for his or her actions and must be aware of the legal implications of practice within the process of role extension. Appropriate training and standards of competence are required for both professional indemnification, provided by the Royal College of Nursing (RCN), and vicarious liability, provided by the individual's employer. There is great variation in nurses' scope of practice, job titles, salaries and standards of training, particularly in the fertility specialism. The United Kingdom Central Council for Nurses, Midwives and Health Visitors (UKCC) is currently piloting a new standard for nurses to attain a 'higher level of practice'. The aim is to provide a generic standard of practice for nurses undertaking an extended role and to overcome many of the inconsistencies experienced by nurses across the UK. Central to any role extension is the delivery of safe care to all patients. Many fertility nurses face new challenges within their practice but require the support of the multidisciplinary team to ensure good standards of patient care. PMID- 11897904 TI - Surgical sperm retrieval: a review of current practice. AB - Although pregnancies were achieved after surgical sperm retrieval and in vitro fertilization 8 years before the introduction of intracytoplasmic sperm injection (ICSI), it is the development of ICSI that has led to the rapid expansion of surgical procedures to obtain sperm from azoospermic males for use in assisted conception cycles. The natural desire for couples to achieve a pregnancy using their own gametes and a national shortage of donor sperm have driven the demand for an expansion of this service. Males who have previously been considered unable to father their own genetic child can now be offered treatment, which, in most cases, will lead to the recovery of sperm for use in treatment. This article considers the development of the techniques available to clinicians and provides an overview of the many treatments (and their acronyms) to assist clinicians unfamiliar with the practicalities of surgical sperm retrieval. In reviewing the current published studies, we also offer some guidelines as to the optimization of the potential future provisions of surgical sperm retrieval treatments for azoospermic males, either secondary to obstruction (particularly after vasectomy) or from non-obstructive causes. PMID- 11897905 TI - A new development in the cryopreservation of sperm. AB - Recent developments in in vitro fertilization have made it necessary to develop better methods for sperm cryopreservation. This article sets out the cryobiological and physical background relevant to sperm cryopreservation together with some new observations on the morphology of sperm in the frozen state. It also provides new data and analysis using a recently developed improved method of cryopreservation. PMID- 11897908 TI - Predicting conception. AB - Infertility is relative. Research into effective treatment must focus on couples with single, identifiable causes for their infertility, and must take into account the effect of chance and time on eventual success. Women with ovulatory infertility, who have no other subfertility factors, can expect normal conception rates when ovulation is restored. Unfortunately, there is no similar simple treatment for male infertility, although complicated and expensive treatment may help. Selection of the couples most likely to benefit from such help will result in the most cost-effective use of scarce resources. Knowledge of duration of infertility and the postcoital test result can help to identify those couples who have a reasonable chance of conception without treatment and those who have virtually no chance without it. PMID- 11897906 TI - Antiphospholipid antibodies and infertility. AB - Antiphospholipid antibodies (aPL) are an established cause of recurrent pregnancy loss. As defective embryonic implantation is a common link between unexplained infertility and recurrent miscarriage, interest has focused on the potential relationship between aPL and implantation failure after in vitro fertilization and embryo transfer (IVF-ET). This review critically examines the published data to determine whether women undergoing IVF-ET should be routinely screened for aPL. Although most studies have reported an increased prevalence of aPL among women undergoing IVF-ET, prospective studies examining the effect of aPL on the outcome of IVF-ET demonstrate that these antibodies do not significantly affect either the implantation or ongoing pregnancy rates. The increased prevalence of aPL among women with infertility is therefore likely to be part of a generalized autoimmune disturbance associated with infertility. Hence routine screening for aPL among women undergoing IVF-ET is not warranted and therapeutic interventions should be used only in well designed randomized controlled trials. PMID- 11897909 TI - Assisted conception in the azoospermic male. AB - The advent of intracytoplasmic sperm injection (ICSI) has offered new solutions for the management of patients with azoospermia. Surgical sperm recovery combined with ICSI has allowed many men with azoospermia to father their own biological children. Azoospermia can be classified as obstructive and non-obstructive, with investigations, management and success rates varying markedly between the two forms. In certain cases of obstructive azoospermia surgical reconstruction remains a viable option, whereas cases with congenital obstruction need to be screened for mutations of the cystic fibrosis gene. In most cases of obstruction sperm can be retrieved from the epididymis using percutaneous epididymal sperm aspiration (PESA). If PESA is unsuccessful, testicular sperm extraction (TESE) is successful in all cases. With non-obstructive azoospermia, the genetic basis has been investigated intensely. Screening for karyotypic abnormalities as well as Y microdeletions is recommended. Irrespective of the histological diagnosis, focal spermatogenesis can be observed in 40-50% of cases using multiple testicular biopsies. PMID- 11897910 TI - Tubal pelvic damage: prediction and prognosis. AB - Tubal pelvic damage is a common cause of infertility, and laparoscopy is the accepted gold standard for its diagnosis. However, laparoscopy is both costly and invasive. Chlamydia is now recognized as the most common cause of tubal pelvic damage. In contrast to laparoscopy, evidence of past chlamydial infection using serology is readily available, and the test is simple and quick to perform. As such, serology can be used as a screening test in infertile women. It is accepted that screening tests may have higher margins of error and may be less accurate than diagnostic tests. Screening is most valuable when detecting a disease for which the treatment is more effective when undertaken at the earliest opportunity. Because there are justified constraints to the indiscriminate use of laparoscopy, there is a need to minimize the number of patients who do not have disease (false positives) who are subjected to this diagnostic investigation. An appropriate Chlamydia antibody titre that would distinguish women at risk of tubal pelvic damage should be determined using diagnostic test analysis and clinical judgement. Identification by serology of women who are likely to have damage would enable these women to undergo a diagnostic test such as laparoscopy sooner, allowing treatment to be provided earlier. However, the severity of tubal pelvic damage varies, and the need to distinguish women with a favourable or unfavourable prognosis after treatment using a simple classification system is discussed. PMID- 11897911 TI - Endometriosis and infertility: the debate continues. AB - A causal relationship between minor endometriosis and infertility or subfertility has not yet been demonstrated, although a significant association is shown by prevalence studies. This article critically reviews the evidence for pituitary ovarian dysfunction as a cause for subfertility in women with minor endometriosis. The lack of fertile controls with endometriosis presents a methodological problem. Group comparison in studies using tubal infertility cases as controls has demonstrated impaired follicular growth, reduced circulating oestradiol concentrations during the preovulatory phase and oestradiol and progesterone during the early luteal phase, and disturbed luteinizing hormone (LH) surge patterns. LH concentration in preovulatory follicular fluid is also reduced, and granulosa cells collected at the same time have impaired steroidogenic capacity in vitro. However, these findings are not consistent in published studies. Significantly lower oocyte fertilization rates (49%) are found compared with controls (69%), even after maximum stimulation with exogenous follicle-stimulating hormone and human chorionic gonadotrophin (52% versus 69%). The implantation rate is also lower (11% versus 13%). An inherent disorder of follicular function seems likely, and LH surge impairment is probably a secondary effect. Impairment of oocyte fertilization would thus contribute substantially to the natural subfertility associated with endometriosis, but in vitro fertilization is still successful as excess numbers of oocytes are available. PMID- 11897913 TI - Reproductive medicine: the ethical issues in the twenty-first century. AB - Reproductive medicine has developed to such an extent that numerous moral questions arise about the boundaries of applications of new reproductive technology. It is possible to imagine a future in which 'designer babies' are created and in which cloning, sex selection and male pregnancy become the instruments of individual desire or social policy. In this article, the concept of 'natural' is explored but rejected as an insufficient moral criterion for deciding these complex questions. A case is made for the criterion of welfare of the child and for the concept of the child as gift rather than product. PMID- 11897912 TI - Polycystic ovary syndrome: not ovarian, not simple, unkind. AB - Polycystic ovary syndrome is one of the most common endocrinopathies worldwide and for many years it was one of the most difficult to elucidate. In the last 10 years, understanding of this condition has increased greatly. It is now clear that polycystic ovary syndrome is not an ovarian disease, rather it is a disorder of intermediary metabolism characterized by insulin resistance. It has also become clear that insulin resistance has implications for general well-being and for the development of novel treatments. PMID- 11897914 TI - Nurses as equals in the multidisciplinary team. AB - For many years nurses have had a high profile both nationally and internationally, but in the past most nurses played only a supporting role. Although the doctor is still recognized as the leader of the clinical team, nurses have become recognized as professionals in their own right, carrying responsibility for their own actions. The clinical and managerial judgement of nurses is respected and increasingly they are given the authority to make policy decisions within a team. However, nurses must take personal responsibility for developing their own leadership and boardroom skills to enable them to make an equal contribution. In this article I reflect on how Professor Michael Hull valued the role of nurses as equal members of the clinical team. In the Centre for Reproductive Medicine in Bristol he paved the way towards making nurses equal. He encouraged nurses to learn and use their knowledge and this in turn developed self-esteem and confidence, which not only made nurses equal but also made them feel equal. PMID- 11897915 TI - Hull and Rutherford classification of infertility. PMID- 11897916 TI - Poor ovarian response to gonadotrophin stimulation the role of adjuvant treatments. AB - Poor ovarian response to gonadotrophin stimulation represents a clinical problem in in vitro fertilization practice. Women showing poor ovarian response are a heterogeneous group, many of whom have a reduced ovarian reserve and consequently a lower pregnancy potential. Various management strategies have been proposed to improve ovarian response to gonadotrophins, but these have met with limited success. Adjuvant treatments aim to potentiate the effect of exogenous follicle stimulating hormone. In separate, randomized, placebo-controlled trials low-dose dexamethasone and aspirin have been shown to reduce the incidence of poor response in an initial stimulation cycle. Preliminary studies using pyridostigmine and L-arginine in established poor responders are encouraging but require confirmation in adequately powered studies. Evidence from randomized controlled trials does not support the use of adjuvant growth hormone or growth hormone-releasing hormone in poor responders without overt growth hormone deficiency. The mechanisms of action of adjuvant treatments require further investigation. PMID- 11897917 TI - Male infertility: tales of progress and frustration. AB - This article describes andrology research, inspired by Professor Michael Hull, to develop sperm function tests and to understand the basic causes of male infertility. No generally acceptable sperm function test has yet been devised. Computer-assisted semen analysis (CASA) proved of limited value in predicting the outcome of in vitro fertilization (IVF) although it was more useful in donor insemination. High intracellular Ca2+ activity, [Ca2+]i, is involved in decreasing motility in cryopreserved sperm. Capacitative calcium entry after depletion of intracellular stores may generate the sustained increase in [Ca2+]i that initiates the acrosome reaction. Our data support the presence of Ca2+ stores as thapsigargin increased [Ca2+]i in sperm in Ca2+ -free medium. Recent observations indicate that cAMP enhances capacitative calcium entry, acting upstream of emptying of the store. Excess reactive oxygen species (ROS) is an important cause of sperm pathology but at low concentrations ROS regulate capacitation. Our evidence shows that ROS are produced by leucocytes present in sperm suspensions. We have been unable to demonstrate that human sperm produce ROS. The relationship between ROS production and lipid peroxidation indicates that sperm from some men are resistant to lipid peroxidation, possibly because of better antioxidant defences. We conclude that the future of andrology lies in the identification of the basic causes of infertility and not in more detailed descriptions of the properties of semen. PMID- 11897918 TI - Cryopreservation, screening and storage of sperm the challenges for the twenty first century. AB - The advent of HIV and the serious nature of the sequelae resulted in a major reassessment of artificial insemination practices in the UK. The development of human semen cryopreservation had enormous impact on reproductive medicine and the availability of cryopreserved quarantined donor semen became a mainstay for the treatment of male infertility in the UK. The regulation and accreditation of assisted reproductive technologies and the introduction of peer-reviewed guidelines have largely standardized clinical and laboratory practice. The introduction of assisted fertilization techniques such as intracytoplasmic sperm injection, testicular sperm retrieval and improved oncology treatments have placed pressure on reproductive biologists and cryobiologists to design and use cryopreservation protocols for the optimum survival of sperm. PMID- 11897919 TI - ReproMED on the Internet today and tomorrow. AB - ReproMED comprises a range of projects applying and researching the role of information technology to support clinical care, education and research in reproductive medicine. This article traces the development of ReproMED on the Internet and concludes by speculating on the future. The project started with a critical evaluation of the infertility information needs of general practitioners and the role of hypertext in meeting those needs, leading to the production of a ReproMED CD-Rom. However, as the project was under development in 1995, Microsoft launched its first web browser amid major expansions of the Internet, thus the project switched to a series of websites and played host to national bodies including the British Fertility Society and the British Andrology Society. After a critical evaluation of the role of the Internet in supporting postgraduate medical education in the southwest region, reproductive medicine training over the Internet was piloted. This pilot was followed by a series of similar training programmes for junior doctors, specialists and general practitioners, leading to the establishment of a formal MSc delivered principally over the Internet. In addition to professional education and research, the ReproMED websites provide patients with extensive interactive information. The future holds many exciting possibilities with rapid technological developments and ever-increasing information needs. PMID- 11897920 TI - Personal recollections of Professor Michael Hull. PMID- 11897921 TI - Clinical research oversight. PMID- 11897922 TI - The generic conundrum. PMID- 11897923 TI - Valdecoxib is more efficacious than rofecoxib in relieving pain associated with oral surgery. AB - Inhibition of the cyclooxygenase (COX)-2 enzyme has been shown previously to reduce pain and inflammation. Valdecoxib is a new highly selective COX-2 inhibitor with a rapid onset of action and significant analgesic properties. This study compared the analgesic efficacy of valdecoxib and rofecoxib in treating postoperative pain in patients undergoing oral surgery. This randomized, double blind, placebo-controlled study compared the efficacy of 40 mg valdecoxib with that of 50 mg rofecoxib and placebo. Efficacy was assessed by the onset of analgesia, pain intensity levels, and pain relief over 24 hours, time-weighted sum of total pain, sum of pain intensity difference, the percentage of patients requiring rescue medication and experiencing regimen failure, and patients' global evaluation. Patients receiving valdecoxib experienced a significantly quicker onset of analgesia, significantly improved pain relief, and lower pain intensity compared with patients receiving rofecoxib and greater satisfaction with their study medication after a single dose. Valdecoxib also demonstrated efficacy that was superior to that of rofecoxib with respect to the percentage of patients requiring rescue medication or experiencing regimen failure (p < or =.05). Valdecoxib, rofecoxib, and placebo were equally well tolerated. This study demonstrates that valdecoxib provides significantly greater analgesic efficacy than rofecoxib in the management of pain after oral surgery. PMID- 11897924 TI - Does heart rate identify sudden death survivors? Assessment of heart rate, QT interval, and heart rate variability. AB - The objective was to test whether the circadian variability of several electrocardiographic variables distinguishes sudden cardiac death survivors from heart disease patients without a history of cardiac arrest and from normal subjects. Heart rate, heart rate variability, and QT interval have been reported to identify survivors of sudden cardiac death. Computer-assisted continuous QT measurement and heart rate variability analysis were performed on 24-hour Holter records for three groups: (1) 14 sudden death survivors; (2) 14 control patients with diagnosis and therapy matched to survivors; and (3) 14 healthy subjects. There were no significant differences in 24-hour mean RR and QT intervals between groups. However, heart rate was significantly different between the three groups at night but not during the day because the expected nighttime decline was markedly blunted in survivors and somewhat blunted in control patients. The QT interval and frequency domain heart rate variability measures followed a similar circadian pattern. The mean QTc was significantly longer in control patients. The QTc had a wide range in all groups, but less in sudden death survivors. Of ten common time and frequency domain heart rate variability indices, only SDANN and SDNN were significantly lower in sudden death survivors. Reduced circadian variation of heart rate, with marked blunting of the nighttime heart rate decline, identifies sudden cardiac death survivors as well as does SDANN and SDNN, and, in contrast to heart rate variability measures, can easily be obtained from a Holter report without complex calculations. PMID- 11897926 TI - Current immunosuppression in liver transplantation. AB - Liver transplantation has emerged from an experimental therapy to a highly successful treatment for end-stage liver disease. The single most important factor in this progression has been the development of effective immunosuppressive regimens. This article will outline the various immunosuppressive agents used in liver transplantation. PMID- 11897925 TI - Comparison of a fixed-dose combination of 40 mg telmisartan plus 12.5 mg hydrochlorothiazide with 40 mg telmisartan in the control of mild to moderate hypertension. AB - This study investigated whether a fixed-dose combination of 40 mg of the angiotensin II antagonist telmisartan plus 12.5 mg of the diuretic hydrochlorothiazide (HCTZ) was superior to 40 mg telmisartan in patients with mild to moderate hypertension who failed to respond adequately to 40 mg telmisartan monotherapy. One hundred forty-six patients were withdrawn before randomization. Nonresponders (n = 327) were double blind and randomized to 40 mg telmisartan + 12.5 mg HCTZ (n = 160) or 40 mg telmisartan (n = 167). After 8 weeks of treatment, 40 mg telmisartan + 12.5 mg HCTZ lowered diastolic blood pressure (DBP) by an additional 3.5 mm Hg (P <.01) and systolic blood pressure (SBP) by 7.4 mm Hg (P <.01) compared with 40 mg telmisartan. Most of the additional effect of the combination was seen after 4 weeks of treatment. At week 8, blood pressure was normalized (SBP <140 mm Hg and DBP <90 mm Hg) in 51.6% of patients on 40 mg telmisartan + 12.5 mg HCTZ compared with 23.5% on 40 mg telmisartan (P <.05). The combination of 40 mg telmisartan + 12.5 mg HCTZ normalized DBP in 64.8% of patients, whereas 40 mg telmisartan normalized DBP in 40.1% (P <.05). SBP decreased by > or =10 mm Hg from baseline in 63.5% of patients receiving the fixed-dose combination compared with 42.6% of those receiving 40 mg telmisartan (P <.05). Both treatments were well tolerated. Adverse events were predominantly mild, transient, and considered unrelated to therapy. These findings indicate that a fixed-dose combination of 40 mg telmisartan + 12.5 mg HCTZ is clinically and statistically superior to 40 mg telmisartan in patients with mild to moderate hypertension failing to respond to 40 mg telmisartan alone. PMID- 11897927 TI - Development of drugs that alter ventricular repolarization. AB - Many drugs are found to alter ventricular repolarization, as manifest by T-wave and U-wave changes on the surface electrocardiogram. These changes have frequently been associated with malignant ventricular arrhythmias. There is no perfectly sensitive and specific way of anticipating such arrhythmias, but some clinical and preclinical screening methods are better than others. The author reviews some of these methods, commenting on some of the regulatory implications. PMID- 11897928 TI - Proarrhythmic effects of adenosine: one decade of clinical data. AB - In 1989, adenosine was introduced into the American clinical setting as an antiarrhythmic drug for the acute management of reentrant supraventricular tachycardia involving the atrioventricular node. During this decade of use, evidence for proarrhythmic effects of the drug have been documented. In addition to the mostly benign transient episodes of atrial fibrillation, several cases of life-threatening ventricular arrhythmias induced by adenosine have been reported. This article summarizes the proarrhythmic effects of adenosine as they were reported in the literature as well as data from the manufacturer files. The causes of these adverse effects of adenosine are analyzed, and factors to be considered before using the drug are discussed. PMID- 11897929 TI - Important role of prodromal viral infections responsible for inhibition of xenobiotic metabolizing enzymes in the pathomechanism of idiopathic Reye's syndrome, Stevens-Johnson syndrome, autoimmune hepatitis, and hepatotoxicity of the therapeutic doses of acetaminophen used in genetically predisposed persons. AB - Upper respiratory tract febrile illnesses caused by various viruses, mycoplasma, chlamydia infections, and/or inflammatory diseases are usually observed a few days to a few (several) weeks before the onset of Reye's syndrome, Stevens Johnson syndrome, autoimmune hepatitis (hepatotropic virus infections), or hepatotoxicity associated with therapeutic administration of acetaminophen in persons with varying degrees of deficits of important enzymatic activity. Activation of systemic host defense mechanisms by inflammatory component(s) results in depression of various induced and constitutive isoforms of cytochrome P-450 mixed-function oxidase system superfamily enzymes in the liver and most other tissues of the body. Because several cytochrome P-450 enzymes activities important for biotransformation of many endogenous and egzogenous substances show considerable variability between individuals, in some genetically predisposed persons, even the administration of therapeutic doses of a drug may result in serious clinical mishaps, if an important concomitant risk factor (eg, acute viral infection) is involved. Several inflammatory cytokines, such as interleukins, transforming growth factor beta1, human hepatocyte growth factor, and lymphotoxin, downregulate gene expression of major cytochrome P-450 enzymes with the specific effects on mRNA levels, protein expression, and enzyme activity observed with a given cytokine varying for each P-450 studied, thus eventually leading to metabolite-mediated adverse drug reactions and immunometallic diseases which sometimes result in tissue injury beyond the site(s) where metabolic bioactivation takes place. On the other hand, it must be emphasized that inhibition of metabolism of several drugs, as well as influence on the concentration and/or ratio of various cytokines in inflamed tissues, may exert beneficial effects in patients with different diseases, thus opening new therapeutic possibilities. Clinically relevant interactions may be exemplified by the effects of some fluoroquinolone antibiotics, such as pefloxacin and ciprofloxacin, which probably have a steroid-sparing effect in some patients with frequently relapsing nephrotic syndrome, and an increased bioavailability of several drugs following concomitant intake with freshly pressed grapefruit juice, eventually caused by inhibition of their metabolism, mediated mainly by CYP3A and specifically inhibited by naturally occurring flavonoids. PMID- 11897930 TI - Treatment of achalasia with botulinum A toxin. AB - Achalasia is an idiopathic neuromuscular disorder of the esophagus which is associated with absence of esophageal peristalsis and incomplete relaxation of a normal or raised lower esophageal sphincter (LES). Dysphagia is the most commonly associated symptom. Conventional therapeutic approaches are directed to reducing LES pressure and include orally-administered smooth muscle relaxants, forceful sphincter dilation with balloon dilators, and open or laparoscopic-assisted myotomy of the LES. Pharmacologic therapies have a low success rate. Forceful dilation has a perforation complication rate of 2% to 5%, and myotomies may precipitate significant gastroesophageal reflux, a complication minimized when a partial fundal wrap is employed simultaneously. In recent years, botulinum toxin, utilized widely as a striated muscle relaxant in managing blepharospasm, anal sphincter spasm, and muscle spasm complicating CVAs, and in smoothening facial wrinkles, has been extended to the management of achalasia on the basis that it impairs smooth muscle responsiveness to acetylcholine. Eighty units of Botox (botulinum toxin) are injected directly into the endoscopically (endoscopic ultrasound techniques may facilitate localization) located LES region (20 units into each of 4 quadrants). Symptom relief lasting 6 months on average is experienced in more than 65% of treated patients, and the complication rate is negligible. This therapeutic option is reserved for patients too ill to undergo any surgical procedure and is most effective when the lower esophageal region is hypertonic. PMID- 11897931 TI - Isoniazid-induced lupus erythematosus presenting with cardiac tamponade. AB - An estimated incidence of drug-induced lupus erythematosus caused by all drugs is 15,000 to 20,000 cases a year, and represents approximately 5 to 10% of the total number of patients with systemic lupus erythematosus. Approximately 22% of the patients treated with isoniazid for a mean of 6 months develop antinuclear antibodies. Isoniazid-induced lupus erythematosus affects either sex equally and the most common presenting feature is arthralgia or arthritis with anemia. Fever and pleuritis occur in approximately half of the cases, and pericarditis in approximately 30% of cases. IgG antibody to the (H2A-H2B)-DNA complex appears specific for the isoniazid-induced lupus erythematosus. The drug-induced lupus presenting with cardiac tamponade is a recognized feature of many drugs such as hydralazine, procainamide, and sulfasalazine. Reported here is a case of isoniazid-induced lupus erythematosus presenting with cardiac tamponade. A 73 year-old man was treated with isoniazid for 8 months at a dose of 300 mg a day. The patient responded to the withdrawal of the isoniazid therapy and placement of a pericardial window. The existing literature on the subject is reviewed. PMID- 11897932 TI - What is quality improvement in the preoperative period? PMID- 11897933 TI - How preoperative assessment programs can be justified financially to hospital administrators. PMID- 11897934 TI - ASA practice guidelines for preanesthetic assessment. PMID- 11897935 TI - Can the risk of postoperative nausea and vomiting be identified and lowered during the preoperative assessment? PMID- 11897936 TI - What should we do with aspirin, NSAIDs, and glycoprotein-receptor inhibitors? PMID- 11897937 TI - Glycemic management of diabetes in the perioperative setting. PMID- 11897938 TI - Why should the anesthesiologist be concerned about endocarditis prophylaxis? PMID- 11897939 TI - Evaluation of the patient with cardiac disease undergoing noncardiac surgery: an update on the original AHA/ACC guidelines. PMID- 11897940 TI - Preoperative noninvasive cardiac testing: which test and why? PMID- 11897941 TI - Perioperative use of beta-blockers: past, present, and future. PMID- 11897942 TI - Transfusion medicine in the preoperative period. PMID- 11897943 TI - Do children need a preoperative assessment that is different from adults? PMID- 11897944 TI - Is there a role for the anesthesiologist in the prenatal clinic? PMID- 11897945 TI - Computer-based preoperative assessment. PMID- 11897946 TI - Holistic issues in the preoperative period. PMID- 11897948 TI - So you want to be a suit. PMID- 11897949 TI - A histologic analysis and three-dimensional reconstruction of the muscle of Riolan. AB - PURPOSE: To describe the anatomic and histologic relations of the muscle of Riolan in the eyelid margin. METHODS: Serial microscopic sections of the eyelid were prepared, digitally scanned, and then reconstructed with computer software to create a 3-dimensional profile of this muscle group in two planes. RESULTS: The muscle of Riolan is a distinct subdivision of striated muscle that is separate from the pretarsal orbicularis muscle. In parasagittal eyelid sections, the muscle appears to be composed of two separate bundles, the pars ciliaris, located anterior to the tarsal plate, and a second smaller bundle, the pars subtarsalis, located posterior to the orifices of the meibomian glands. Coronal sections, however, demonstrate numerous muscle fibers that traverse the tarsus, connecting the two muscle groups that we describe for the first time as the pars fascicularis. CONCLUSIONS: The three muscle subdivisions are therefore physically joined together and appear to act as a single functional entity that should be collectively referred to as the muscle of Riolan. PMID- 11897950 TI - Eyelid lymphatics II: a search for drainage patterns in the monkey and correlations with human lymphatics. AB - PURPOSE: To study the lymphatic drainage of the cynomolgus monkey through the use of lymphoscintigraphy. METHODS: Lymphoscintigraphy with 500 microCi of 99mTechnetium sulfur colloid injected at specific sites around the eyelids was performed with five cynomolgus monkeys in lateral and ventral positions. RESULTS: Lymphoscintigraphy of the monkey eyelid and periocular tissue revealed lymphatic drainage to the parotid lymph nodes from the entire upper eyelid, medial canthus, and lateral lower eyelid and drainage to the submandibular lymph nodes from the medial and central lower eyelid. In addition to draining to the parotid lymph nodes, the central upper eyelid was also seen to drain to the submandibular lymph nodes. CONCLUSIONS: Lymphoscintigraphy of the cynomolgus monkey eyelids reveals discrete lymphatic drainage pathways for the upper and lower eyelids and a dual pathway for the central upper eyelid. Future studies will help to clarify the lymphatic drainage pathways of human eyelids. PMID- 11897951 TI - Qualitative perfusion imaging of the human optic nerve. AB - PURPOSE: Standard methods for the evaluation of human optic nerve perfusion provide limited information. In this pilot study, the authors investigated the feasibility of qualitative perfusion imaging, a recently developed neuroradiologic technique, as a method of assessing human intraorbital optic nerve blood flow. METHODS: Qualitative perfusion imaging (based on magnetic resonance fast spin-echo sequences) was used to study the optic nerves of 7 healthy volunteers and 5 patients with known optic nerve disease. Data regarding both study subject background and alteration in optic nerve signal intensity were statistically analyzed. RESULTS: Control group subjects were significantly younger than study group subjects. No significant differences in optic nerve signal patterns were found within the control group. Comparison of patients with optic neuropathy against the normal composite revealed substantial differences in enhancement characteristics. CONCLUSIONS: Qualitative perfusion imaging of the human optic nerve is feasible and may serve as the basis for more advanced neuroradiologic studies of optic nerve blood flow abnormalities. PMID- 11897952 TI - Computed tomographic findings of distensible orbital venous anomalies. AB - PURPOSE: To describe the CT findings of distensible orbital venous anomalies. METHODS: We reviewed the records of 17 consecutive patients who met the imaging criteria for orbital venous anomalies. We analyzed the clinical data and the CT scans qualitatively and quantitatively. RESULTS: Distensible venous anomalies usually occupied both intraconal and extraconal spaces of the orbit (65%). The contour of the mass was ovoid or fusiform in 3 of the orbits (18%) and tubular or lobular in 14 orbits (82%). The optic nerve was encased by the mass in 8 orbits. Changes in the adjacent bone were detected in 9 orbits. The average volume of the venous anomalies was 1.1 mL in the axial scan and 5.7 mL in the coronal scan. The patients who showed clinically evident exophthalmos on the Valsalva maneuver had larger volumes than those who did not. CONCLUSIONS: The orbital structures adjacent to the distensible orbital venous anomalies were vulnerable to intermittent compression. CT scans, including both postcontrast axial and coronal images, were useful in demonstrating the presence of distensible venous anomalies, even in cases with no apparent exophthalmos on the Valsalva maneuver. PMID- 11897953 TI - Helical computed tomographic dacryocystography with three-dimensional reconstruction: a new view of the lacrimal drainage system. AB - PURPOSE: To evaluate the utility of a new diagnostic radiologic technique for anatomic evaluation of the lacrimal outflow system in patients with epiphora and to delineate anatomic variations in the lacrimal systems of patients with both patent and obstructed systems. METHODS: This study retrospectively reviewed clinical and radiologic data in a series of 30 patients with epiphora who underwent our radiologic protocol. Imaging included fluoroscopic dacryocystography followed by helical CT dacryocystography after injection of lacrimal system contrast. Axial CT data was three-dimensionally reconstructed and rotated for viewing of images in multiple projections. Mean axial cross-sectional areas of the lacrimal sac and duct were determined. RESULTS: Sixty lacrimal systems in 30 patients were clinically and radiologically evaluated. The average mean cross-sectional area of the lacrimal sac and duct in the setting of complete obstruction was 0.153 cm2 and was statistically significantly larger (p=0.0286) when compared with average mean cross-sectional areas in unobstructed (0.045 cm2) and partially obstructed (0.052 cm2) lacrimal systems and were associated with lacrimal system dilation proximal to the level of obstruction. The difference in average mean cross-sectional area between patients with unobstructed and partially obstructed systems was not statistically significant. A number of lacrimal system abnormalities were noted in our series, including obstructions at various levels of the lacrimal outflow system, lacrimal sac masses, sinusitis, sarcoidosis, sinus carcinoma, and failed dacryocystorhinostomy. Twenty-three lacrimal systems were believed to be radiographically normal. Radiologic findings altered surgical treatment in 10 of 30 patients in this series. CONCLUSIONS: This relatively safe and well-tolerated radiologic technique provides detailed imaging of the lacrimal outflow system and surrounding structures. The information obtained from this technique may be helpful in clinical and surgical decision making. PMID- 11897954 TI - Medial tarsal suspension: a method of elevating the medial lower eyelid. AB - PURPOSE: When attempting to elevate the lower eyelid for any reason, medial elevation is the most difficult to attain. Medial canthal tendon tightening creates mostly horizontal tension and contributes little vertical vector. We present a technique for applying a lifting force to the medial end of the eyelid: medial tarsal suspension. METHODS: The technique to suspend the medial lower eyelid tarsal plate to the superior orbital rim periosteum is described. The procedure, medial tarsal suspension (MTS), was performed on 38 lower lids of 24 patients. Adjunctive procedures, most commonly lateral canthal sling, were performed on 66% of the lids at the time of the initial medial tarsal suspension. The patients ranged in age from 29 years to 84 years. All had medial lower eyelid retraction, with facial nerve palsy, Graves eye disease, involutional lower eyelid retraction, and forms of muscular dystrophy the commonest etiologies. RESULTS: Thirty-one (82%) of the 38 MTS procedures were successful. There was no unifying factor among the seven failed procedures in five lids of five patients. Three of the five patients, including two who were operated on twice, ultimately had a successful MTS. This procedure was not repeated on the other two failed patients. Range of follow-up was 9 months to 5.6 years, with a mean of 3.7 years. The mean elevation of the central lower eyelid was 1.6 mm in the successful cases. CONCLUSIONS: Medial tarsal suspension is an effective way to elevate the medial end of the lower eyelid. PMID- 11897956 TI - Corneal degeneration following bicanalicular silicone intubation. AB - PURPOSE: A case of bilateral arcus-like corneal degeneration in the medial limbal regions following bilateral bicanalicular silicone intubation is reported. METHODS: Case report. RESULTS: Bilateral arcus-like corneal opacifications were observed in the medial limbal regions of a 33-year-old woman 3 months after the bilateral silicone intubation with dacryocystorhinostomy. No clinical signs of infection and direct contact of the silicone tubes with those regions of the corneas were observed. Routine laboratory tests revealed no abnormalities, and serum lipoprotein composition was normal. CONCLUSIONS: Silicone tubing appeared to be the precipitating factor of the corneal opacifications that occurred in this patient. PMID- 11897955 TI - Advancement flaps for large defects of the eyebrow, glabella, forehead, and temple. AB - PURPOSE: To describe a system for reconstruction of large defects of the eyebrow, glabella, forehead, and temple. The system maximizes the use of direct approximation and advancement flaps before resorting to less aesthetic techniques. METHODS: This was a retrospective cohort study drawn from approximately 70 patients with post-Mohs defects of the eyebrow, glabella, forehead, and temple. Surgical intervention involved the graded application of direct approximation, horizontally oriented advancement flaps, rotational flaps, and free skin grafts. The selection of individual and combined techniques was based on defect area and depth, elasticity of adjacent tissues, and relations of the defect to the neighboring eyebrows and hairlines. RESULTS: Reconstructive techniques applied to defects of the eyebrow, glabella, forehead, and temple can be arranged in an incremental scale that provides progressively more tissue but at an escalating aesthetic cost. The usual defect size limits for direct approximation and advancement flaps can be expanded. CONCLUSIONS: To avoid the limitations of large rotational flaps and skin grafts in this region, maximal use of direct approximation and advancement flaps is recommended. PMID- 11897957 TI - Bilateral congenital lacrimal anlage ducts (lacrimal fistula) in a Patient With the VACTERL association. AB - PURPOSE: To present a case of bilateral congenital lacrimal anlage ducts in a patient with the VACTERL (vertebral anomalies, anal atresia, cardiac malformations, tracheoesophageal fistula, renal anomalies, and limb anomalies) association. METHODS: Case report. RESULTS: A 19-year-old man presented with intermittent facial swelling. Examination revealed bilateral lacrimal anlage ducts. The patient underwent excision of the ducts with silicone intubation. CONCLUSIONS: We add the VACTERL association to the list of systemic conditions associated with lacrimal anlage ducts. PMID- 11897958 TI - Cicatricial upper eyelid retraction in encephalocraniocutaneous lipomatosis: a report of two cases and review of literature. AB - PURPOSE: To describe two patients with encephalocraniocutaneous lipomatosis (ECCL) and to review the literature on this disorder. METHODS: Brain and orbit CT scans were performed on two patients with ECCL. Both patients were examined by the same ophthalmologist and neurologist during at least a 2-year follow-up period. RESULTS: Unilateral skull hamartomas, intracranial abnormalities, epibulbar choristomas, and ocular adnexal changes including a specific form of cicatricial upper eyelid retraction were present in both patients. CONCLUSIONS: ECCL is a special form of oculocerebrocutaneous disease that has significant adnexal findings that are essential for the diagnosis of this rare disorder. PMID- 11897959 TI - Ocular findings in cutis laxa acquisita. AB - PURPOSE: To report ocular findings in a patient with cutis laxa acquisita. METHODS: Case report RESULTS: A 44-year-old man complained of excessively loose skin for 4 years and had no family history of skin disease. Dermatological examination showed lax and wrinkled skin all over his body. He had bilateral subconjunctival fat prolapse and dermatochalasis. CONCLUSIONS: This case of acquired form of cutis laxa is noteworthy because subconjunctival fat prolapse and dermatochalasis are uncommon at this relatively young age. PMID- 11897960 TI - Leiomyosarcoma of the esophagus metastatic to the eyelid: a clinicopathologic report. AB - PURPOSE: To report a case of gastrointestinal sarcoma with leiomyosarcomatous differentiation metastatic to the eyelid. METHODS: Clinical data including a comprehensive ophthalmologic examination, histologic findings, a bone scan, and hospital records were reviewed. RESULTS: A 56-year-old woman with a history of esophageal cancer had a rapidly growing lesion on her right upper eyelid that was initially treated as a chalazion. A biopsy specimen of the lesion was consistent with a gastrointestinal sarcoma with leiomyosarcomatous differentiation. The esophageal tumor was reclassified after histopathologic evaluation of the eyelid specimen. Bony metastasis developed soon after excision of the eyelid lesion. The patient died 3 months after proper diagnosis of the eyelid lesion. CONCLUSIONS: This is a rare case of a metastatic leiomyosarcoma of the eyelid. To our knowledge, only two other cases of eyelid leiomyosarcoma have been described in the literature. The importance of correct histopathologic diagnosis of eyelid lesions is underscored in this case report. PMID- 11897961 TI - Unmasking of an orbital dermolipoma following aesthetic facial surgery. PMID- 11897963 TI - Superomedial lid crease approach to the medial intraconal space. PMID- 11897962 TI - MEN IIB and ganglioneuromatosis. PMID- 11897965 TI - Toxicologic aspects of heroin substitution treatment. AB - Heroin abuse is an international problem with which all countries must continually cope. Many countries have implemented heroin substitution therapy as an effective means of decreasing illicit heroin use, crime, HIV risk, and death, and in improving employment and social adjustment. Although methadone is the most commonly used medication for heroin substitution, other agonists in current use include levomethadyl acetate (LAAM), buprenorphine, and pharmaceutical-grade heroin. This report reviews toxicologic issues that arise in these programs. A broad array of testing methodologies are available that allow selection of on site testing or laboratory-based methodology. Urine specimens may be monitored for nonprescribed drugs on a qualitative or semiquantitative basis. Methods for differentiating opiate sources by urinalysis have been proposed to distinguish poppy seed consumption from heroin abuse and for distinguishing pharmaceutical grade heroin from illicit heroin. Therapeutic drug monitoring for methadone in plasma continues to be evaluated for use in establishing adequate dosing and detecting diversion, and new methods have been devised for measurement of the optical isomers of methadone in plasma. Biologic specimens, in addition to plasma and urine, have been evaluated for use in drug monitoring, including sweat, hair, and oral fluid, with promising results. Overall, the many recent developments in testing methodology provide more effective means to assess patients in heroin substitution programs and should contribute to improvements in public health. PMID- 11897966 TI - Postmortem drug analysis: analytical and toxicological aspects. AB - Publications focusing on the analysis of postmortem specimens for the presence of drugs were reviewed with particular reference to systematic toxicological analysis. Specimens included blood, liver, other solid specimens, and fly larvae. Extraction techniques published during the past 10 years most commonly used traditional solvent extraction techniques. High-performance liquid chromatography coupled to multichannel wavelength detection was most commonly used, which would easily lend itself to liquid chromatography-mass spectrometry. There were few practical differences in the assays validated for a range of postmortem specimens to those in other forms of forensic toxicology, unless substantially decomposed tissue was used. When putrefied specimens were analyzed, a back-extraction or other form of specimen cleanup was recommended to reduce interfering substances. Many immunoassays designed for urine have been adapted for use in blood and tissue homogenates. Immunoassays designed for blood analysis, however, are likely to have more useful cutoff values than immunoassays optimized for urine testing. Postmortem specimens provide less stability for a number of drugs than other types of specimens. This is particularly a problem for cocaine, heroin, and some antidepressants, antipsychotics, and benzodiazepines. A number of artifacts occur postmortem, which affects the concentration of drug in specimens. This includes postmortem redistribution for drugs with a high tissue concentration relative to blood. Consequently, the likely extent of any change in concentration is relevant to the interpretation of doses and drug effects. PMID- 11897967 TI - Drugs of abuse monitoring in blood for control of driving under the influence of drugs. AB - Driving under the influence of drugs is an issue of growing concern in the industrialized countries as a risk and a cause for road accidents. In forensic toxicology, the increasing number of samples for determination of drugs in blood is mainly due to zero-tolerance laws in several countries and well-trained police officers who can better recognize drivers under the influence of drugs of abuse. This review describes procedures for detection of the following drugs of abuse in whole blood, plasma, and serum: amphetamine, methamphetamine, 3,4-methylenedioxy methamphetamine (MDMA), N-ethyl-3, 4-methylenedioxyamphetamine (MDEA), 3,4 methylenedioxyamphetamine (MDA), cannabinoids (delta-9-tetrahydrocannabinol [THC], 11-hydroxy-delta-9-THC, 11-nor-9-carboxy-delta-9-THC), cocaine, benzoylecgonine, ecgonine methyl ester, cocaethylene, the opiates (heroin, 6 monoacetylmorphine, morphine, or codeine), and methadone as well as gamma hydroxybutyric acid (GHB), lysergic acid diethylamide (LSD), phencyclidine (PCP), and psilocybin/psilocin. For many of the analytes, sensitive immunologic methods for screening are available. Gas chromatography-mass spectrometry (GC-MS) is still the state-of-the-art method for confirmatory analysis or for screening and confirmation in one step. Liquid chromatography-mass spectrometry (LC-MS) procedures for such purposes are also included in this review. Basic data about the biosample assayed, internal standard, workup, GC or LC column and mobile phase, detection mode, reference data, and validation data of each procedure are summarized in two tables. PMID- 11897968 TI - Progress of capillary electrophoresis in therapeutic drug monitoring and clinical and forensic toxicology. AB - During the past decade, capillary electrophoresis (CE) emerged as a promising, effective, and economical approach for the analysis of licit and illicit drugs and their metabolites in biologic samples. This review provides an overview of the principles of CE, the features of CE instrumentation, and the key aspects of CE-based drug assays that were developed for therapeutic drug monitoring (TDM), clinical and forensic toxicology, and assessment of drug metabolism and pharmacokinetics. CE performed in fused-silica capillaries has sufficiently matured and can thus be applied routinely, whereas chip-based instrumentation comprising fully integrated assays is still in development. Despite the attractive advantages of electrokinetic capillary technology, relatively few CE based assays for TDM and for drug screening of clinical and forensic interest have been adopted in the routine arena. The lack of complete systems designed for unattended operation, the reluctance of bioanalysts to replace a satisfactory existing method, and tight budgets are believed to have hindered the widespread replacement of older (mainly chromatographic) technology. Another limitation of CE is that this technique is somewhat less sensitive than other analytic techniques used for drug analysis in biologic fluids. New instrumental developments featuring user-friendly software and the introduction of assay kits, however, should increase the number of validated CE drug tests becoming used on a routine basis. PMID- 11897969 TI - Clinical toxicologic implications of ethylene glycol and glycolic acid poisoning. AB - Metabolic pathways have been elucidated for various chemical and solvent exposures in humans. Clinical laboratory analyses in most chemical and solvent exposures are directed toward identification and quantitation of unchanged substance in serum or whole blood. For example, most laboratories routinely screen for unchanged ethylene glycol in suspected poisonings and quantitate ethylene glycol in positive cases even though toxicity from ethylene glycol exposure (including central nervous system depression, acute renal failure, and elevated anion gap metabolic acidosis) is primarily caused by one metabolite glycolic acid. One objective of this manuscript is to describe the authors' clinical experience with glycolic acid analysis in ethylene glycol human poisonings. Recommended clinical laboratory tests for small hospitals and toxicology reference laboratories are presented to rule out or confirm ethylene glycol exposure. Another concern with laboratory support in ethylene glycol poisoning is correct identification of ethylene glycol because analysis of this substance is often problematic. In one case laboratories incorrectly identified an organic acid from an inherited metabolic disease as ethylene glycol, and in another case the intentional ethylene glycol poisoning of an infant was determined to be the results of an endogenous organic acid. The most robust analytical methods for determining ethylene glycol and glycolic acid are chromatographic methods. Ideally, screening methods for ethylene glycol should be confirmed by another method based on a different principle of analysis or include simultaneous metabolite analysis (glycolic acid). In centers where several ethylene glycol cases present annually, toxicology laboratories supporting these centers should incorporate glycolic acid monitoring in their ethylene glycol screening programs and include analysis of both ethylene glycol and glycolic acid during treatment (hemodialysis) in all confirmed poisonings. Measurement of glycolic acid provides important diagnostic and prognostic information that one cannot correlate with the amount of ethylene glycol in serum or whole blood. PMID- 11897970 TI - Use of alternative specimens: drugs of abuse in saliva and doping agents in hair. AB - It is generally accepted that chemical testing of biologic fluids is the most objective means of diagnosis of drug use. The presence of a drug analyte in a biologic specimen can be used to document exposure. The standard for drug testing in toxicology is an immunoassay screen conducted on a urine sample, followed by confirmation by gas chromatography with mass spectrometric detection. In recent years, remarkable advances in sensitive analytic techniques have enabled the analysis of drugs in unconventional biologic specimens such as saliva or hair. The aim of this review is to document the current status of drugs of abuse testing in saliva and some doping agents in hair. The influence on drug concentration of the procedure of saliva sampling is described. Screening procedures along with specific methods are reviewed for the determination of amphetamines, cannabis, cocaine, and opiates in saliva. Before an extensive review on the detection of anabolics, corticosteroids, and beta-adrenergic stimulants in hair, the place of this specimen in doping control is discussed, with a focus on the potential problems of this new technology. PMID- 11897971 TI - Role of gas chromatography-mass spectrometry with negative ion chemical ionization in clinical and forensic toxicology, doping control, and biomonitoring. AB - This paper reviews procedures for the detection or quantification of drugs, pesticides, pollutants, and/or their metabolites relevant to clinical and forensic toxicology, doping control, or biomonitoring using gas chromatography mass spectrometry with negative ion chemical ionization (GC-MS-NICI). Papers written in English between 1995 and 2000 are reviewed. Procedures are included for the analysis of the following halogen-containing or derivatizable compounds in common biosamples, such as whole blood, plasma, or urine, and in alternative matrices such as sweat, hair, bone, or muscle samples of humans or rats: benzodiazepines, cannabinoids, opioids, acetylsalicylic acid, angiotensin converting enzyme inhibitors, ketoprofen, methylphenidate enantiomers, tegafur, zacopride, anabolic steroids, chlorophenols, chlorpyrifos, hexachlorocyclohexanes, organochlorines, and polychlorinated biphenyls. The principal information on each procedure is summarized in three tables to facilitate the selection of a method suitable for a specific analytic problem. PMID- 11897972 TI - Progress of liquid chromatography-mass spectrometry in clinical and forensic toxicology. AB - The use of liquid chromatography-mass spectrometry (LC-MS) has recently exploded in various analytic fields, including toxicology and therapeutic drug monitoring (although still far behind pharmacokinetics). There is no doubt that LC-MS is currently competing with gas chromatography (GC)-MS for the status of the reference analytic technique in toxicology. This review presents, for the nonspecialist reader, the principles, advantages, and drawbacks of LC-MS systems using atmospheric pressure interfaces. It also gives an overview of the analytic methods for xenobiotics that could be set up with these instruments for clinical or forensic toxicology. In particular, as far as quantitative techniques are concerned, this review tries to underline the large number and variety of drugs or classes of drugs (drugs of abuse, therapeutic drugs) or toxic compounds (e.g., pesticides) that can be readily determined using such instruments, the respective merits of the different ionization sources, and the improvements brought about by tandem MS. It also discusses new applications of LC-MS in the field of toxicology, such as "general unknown" screening procedures and mass spectral libraries using LC-atmospheric pressure ionization (API)-MS or MS-MS, presenting the different solutions proposed to overcome the naturally low fragmentation power of API sources. Finally, the opportunities afforded by the most recent or proposed instrument designs are addressed. PMID- 11897973 TI - Toxicokinetics of amphetamines: metabolism and toxicokinetic data of designer drugs, amphetamine, methamphetamine, and their N-alkyl derivatives. AB - This paper reviews the toxicokinetics of amphetamines. The designer drugs MDA (methylenedioxy-amphetamine, R,S-1-(3;,4;-methylenedioxyphenyl)2-propanamine), MDMA (R,S-methylenedioxymethamphetamine), and MDE (R,S methylenedioxyethylamphetamine), as well as BDB (benzodioxolylbutanamine; R,S-1 (1;,3;-benzodioxol-5;-yl)-2-butanamine or R,S-1-(3;,4;-methylenedioxyphenyl)-2 butanamine) and MBDB (R,S-N-methyl-benzodioxolylbutanamine), were taken into consideration, as were the following N-alkylated amphetamine derivatives: amphetaminil, benzphetamine, clobenzorex, dimethylamphetamine, ethylamphetamine, famprofazone, fencamine, fenethylline, fenproporex, furfenorex, mefenorex, mesocarb, methamphetamine, prenylamine, and selegiline. English-language publications from 1995 to 2000 were reviewed. Papers describing identification of metabolites or cytochrome P450 isoenzyme-dependent metabolism and papers containing pharmacokinetic/toxicokinetic data were considered and summarized. The implications of toxicokinetics for toxicologic assessment or for interpretation in forensic cases are discussed. PMID- 11897974 TI - Role of chiral chromatography in therapeutic drug monitoring and in clinical and forensic toxicology. AB - Advances in chiral chromatographic separations have given pharmacologists and toxicologists the tools to examine unexpected clinical results involving chiral drugs. The ability to unravel complex phenomena associated with drug transport and drug metabolism is presented in this manuscript. The relation between the chirality of the drug mefloquine and the intracellular concentrations of the drug cyclosporine is illustrated by examining the effect of the enantiomers of mefloquine on the transport activity of P-glycoprotein (Pgp). These studies were conducted using a liquid chromatographic column containing immobilized Pgp. The results demonstrated that (+)-mefloquine competitively displaced the Pgp substrate cyclosporine whereas (-)-mefloquine had no effect on cyclosporine-Pgp binding. The data suggest that cyclosporine cellular and CNS concentrations can be increased through the concomitant administration of (+)-mefloquine. The use of chirality in clinical and forensic situations is also illustrated by the metabolism of the enantiomers of ketamine (KET). The plasma concentrations of (+) KET and (-)-KET and the norketamine metabolites (+)-NK and (-)-NK were measured in rat plasma using enantioselective gas chromatography. The separations were accomplished using a gas chromatography chiral stationary phase based on beta cyclodextrin. The pharmacokinetic profiles of (+)-, (-)-KET and (+)-, (-)-NK were determined in control and protein-calorie malnourished (PCM) rats to determine the effect of PCM on ketamine metabolism and clearance. The results indicate that PCM produced a significant and stereoselective decrease in KET and NK metabolism. The data suggest that the effects of environmental factors (smoking, alcohol use, diet) and drug interactions (coadministered agents) can be measured using the changes in stereochemical metabolic and pharmacokinetic patterns of KET and similar drugs. PMID- 11897975 TI - Evaluation of REMEDi HS in the diagnosis of dimethoate poisoning. AB - The authors describe the evaluation of an automated rapid emergency drug profiling system (REMEDi HS), which is designed for clinical toxicology and therapeutic drug monitoring, in a severe case of dimethoate poisoning. Successful qualitative and quantitative determination of dimethoate in diluted serum samples was achieved. The authors found that dimethoate serum concentrations were properly measured with the REMEDi method and well correlated with gas chromatography-mass spectrometry. The results provide evidence that besides screening and rapid estimation of drug concentrations in human fluids, this system is a suitable tool in cases of symptomatic dimethoate poisoning. PMID- 11897976 TI - Simultaneous determination of the six HIV protease inhibitors (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir) plus M8 nelfinavir metabolite and the nonnucleoside reverse transcription inhibitor efavirenz in human plasma by solid-phase extraction and column liquid chromatography. AB - A sensitive and selective liquid chromatographic assay has been developed for the determination of the six currently protease inhibitors approved by the U.S. Food & Drug Administration (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir) plus the M8 active metabolite of nelfinavir and the nonnucleoside reverse transcription inhibitor efavirenz in a single run. Pretreatment of 1-mL plasma sample spiked with internal standard was made by a solid-phase extraction procedure using a polymeric reversed-phase sorbent. Liquid chromatography was performed using a narrow-bore C18 reversed-phase column and gradient elution. Double ultraviolet detection at 265 nm (amprenavir) and at 210 nm (all other assayed drugs and internal standard) was used. Calibration curves were linear in the range 25 to 10,000 ng/mL, and the assay has been validated over the range 25 to 5,000 ng/mL. Average accuracies at four concentrations were in the range 92.4% to 103.0% and 94.4% to 103.0% for within-day and between-day, respectively, and the coefficients of variation were less than 8%. Mean absolute recoveries varied from 72.8% (ritonavir) to 93.7% (indinavir). No metabolite of the protease inhibitors was found to coelute with the drugs of interest or with the internal standard. At this time, among the tested drugs, especially all the currently licensed nucleosides and the other nonnucleoside reverse transcription inhibitor nevirapine that can be used in combination with the protease inhibitors, none was found to interfere with the assay. PMID- 11897977 TI - Pharmacokinetics and protein binding of mycophenolic acid in stable lung transplant recipients. AB - Mycophenolate mofetil (MMF) use is increasing in solid organ transplantation. Mycophenolic acid (MPA), the active metabolite of MMF, is highly protein bound and only free MPA is pharmacologically active. The average MPA free fraction in healthy adult individuals, stable renal transplant recipients, and heart transplant recipients is approximately 2 to 3%. However, no data are currently available on MPA protein binding in stable lung transplant recipients and little is known regarding MPA's pharmacokinetic characteristics after lung transplantation. The purpose of this study was to characterize the pharmacokinetic profile and protein binding of MPA in this patient population. Seven patients were entered into the study. On administration of a steady-state morning MMF dose, blood samples were collected at 0, 1, 2, 3, 4, 5, 6, 8, 9, 10, and 12 hours post-dose. Total MPA concentrations were measured by a validated HPLC method with UV detection and followed by ultrafiltration of pooled samples for free MPA concentrations. Area under the curve (AUC), peak concentration (Cmax), time to peak concentration (Tmax), trough concentration (Cmin), free fraction (f), and free MPA AUC were calculated by traditional pharmacokinetic methods. Patient characteristics included; 3 males and 4 females, an average of 4.4 years post-lung transplant (range, 0.3-11.5 yr), mean (+/- SD) age of 50 +/- 10 years and weight 69 +/- 20 kg. Mean albumin concentration was 37 +/- 3 g/L and serum creatinine was 142 +/- 49 micromol/L. All patients were on cyclosporine and prednisone. MMF dosage ranged from 1 to 3 g daily (35.5 +/- 14.1 mg/kg/d; range, 15.2-60.0 mg/kg/d). Mean (+/- SD) AUC was 45.78 +/- 18.35 microg.h/mL (range, 16.56-74.22 microg.h/mL), Cmax was 17.37 +/- 7.69 microg/mL (range, 4.92-26.63 microg/mL), Tmax was 1.2 +/- 0.4 hours (range, 1.0-2.0 h), Cmin was 3.12 +/- 1.41 microg/mL (range, 1.47-4.82 microg/mL), f was 2.90 +/- 0.56% (range, 2.00-3.40%), and free MPA AUC was 1.29 +/- 0.50 microg.h/mL (range, 0.54-1.88 microg.h/mL). This is the first study to determine these pharmacokinetic characteristics of MPA in the lung transplant population. Further studies should focus on identification of MMF dosing strategies that optimize immunosuppressive efficacy and minimize toxicity in lung allograft recipients. PMID- 11897978 TI - Development of population pharmacokinetic models and optimal sampling times for ibuprofen tablet and suspension formulations in children with cystic fibrosis. AB - High-dose ibuprofen therapy has demonstrated to slow deterioration in pulmonary function in children with cystic fibrosis with mild lung disease. Therapeutic drug monitoring has been recommended to maintain peak concentrations within the range of 50 to 100 mg/L to ensure efficacy. Current methods for dosage individualization are based on dose proportionality using visual inspection of the peak concentration; however, because of interpatient variability in the absorption of the various formulations this method may result in incorrect assessments of the peak concentration achieved. Maximum a posteriori Bayesian analysis (MAP-B) has proven to be a useful and precise method of individualizing the dose of aminoglycosides but requires a description of the structural model. In this study we performed parametric population modeling analysis on plasma concentrations of ibuprofen after single doses of 20 to 30-mg/kg tablet or suspension in children with cystic fibrosis. Patients evaluated in this study were part of a single dose pharmacokinetic study that has been published previously. A one-compartment model with first order absorption and a lag time best described the data. The pharmacokinetic parameters differed significantly depending on the formulation administered. D-optimal sampling times for the suspension and tablet formulations are 0, 0.25 to 0.5, 1, and 3 to 4 hours and 0, 0.25 to 0.5, 1 to 1.5, and 5 hours respectively. Use of MAP-B analysis performed with the 4 d-optimal sampling strategy resulted in accurate and precise estimates of the pharmacokinetic parameters when compared with maximum likelihood analysis using the complete plasma concentrations data set. Further studies are needed to evaluate the performance of these models and the impact on patient outcomes. PMID- 11897980 TI - Endotoxin exposure in allergy and asthma: reconciling a paradox. AB - Well-established evidence links endotoxin exposure, especially in the workplace, to airways disease. Endotoxin can increase disease severity by acting as a natural adjuvant to augment asthma and atopic inflammation. Recent studies suggest that it can even act on its own, causing a distinct endotoxic form of asthma. Other studies, however, contradict the paradigm that endotoxin's influence is solely a negative one. Epidemiologic associations of environmental endotoxin exposure with allergy and asthma prevention are consistent with hygiene hypothesis associations of other microbial exposures or infections with a lower incidence of atopic disease. Currently, microbe-derived products are being developed as potential therapies for allergy and asthma. Thus it is an ideal time to consider endotoxin as a prototype of a natural intervention with microbial components. Nature's ongoing experiment with endotoxin can provide clues for the development of effective and safe microbe-based products for disease treatment and prevention. This article will discuss (1) conventional paradigms in which endotoxin-induced immune modulation by T(H)1-type induction leads to mitigation of T(H)2-type immune development, allergen sensitization, and atopic inflammation; (2) newer concepts of T(H)1-type immune responses that may provide additional asthma-protective effects by preventing airways remodeling; (3) home and environmental features that significantly contribute to endotoxin exposure; (4) different aspects of asthma mediated by endotoxin exposure; and (5) how to understand endotoxin's paradoxical nature of serving as both friend and foe. PMID- 11897982 TI - Tacrolimus ointment: its place in the therapy of atopic dermatitis. PMID- 11897981 TI - The cellular biology of eosinophil eicosanoid formation and function. AB - Eosinophils are capable of generating eicosanoid derivatives of arachidonic acid by means of cyclooxygenase and the 5- and 15-lipoxygenase (LO) pathways. Moreover, eosinophils, because of their expression of leukotriene (LT) C(4) synthase, are a major source of 5-LO-derived cysteinyl LTs, which are potent paracrine mediators of bronchial obstruction and inflammation pertinent to asthma. The regulation of eicosanoid formation within eosinophils involves activation of key enzymes at specific intracellular sites. Calcium ionophore elicited translocation of 5-LO to the membranes of the nuclear envelope is associated with LTC(4) formation. In addition, lipid bodies, the formation of which is initiated by specific receptor-mediated signaling pathways, are sites of cyclooxygenase- and LO-pathway eicosanoid formation. Newly formed LTC(4) can be immunolocalized at perinuclear membranes in ionophore-activated eosinophils and at lipid bodies in CCR3 agonist (eg, eotaxin) chemokine-stimulated eosinophils. The local generation of eicosanoids at distinct sites within eosinophils may be important for the roles of these eicosanoids, both as paracrine mediators pertinent to inflammation and as intracrine signal-transducing mediators that help regulate cellular responses of eosinophils. PMID- 11897983 TI - The IL-5 receptor on human bronchus selectively primes for hyperresponsiveness. AB - BACKGROUND: The role of IL-5-induced eosinophilia in airway hyperresponsiveness has been questioned. In addition, eosinophil-independent IL-5-induced airway hyperresponsiveness has been demonstrated in animals. OBJECTIVE: In this study, IL-5 was investigated for direct effects on human bronchial responsiveness. METHODS: Human muscle preparations were isolated from organ donor and surgical tissue. Bronchus, jejunum, and saphenous vein were incubated for 24 hours in vitro with recombinant human (rh) IL-5. Contractility to acetylcholine (bronchus, jejunum) and phenylephrine (saphenous vein) was then investigated. RT-PCR was used to evaluate IL-5 receptor alpha (IL-5R(alpha)) expression in various tissues and to assess bronchus and saphenous vein eosinophils through use of CCR3 expression. RESULTS: rhIL-5 primed bronchus for an exaggerated contraction to acetylcholine. The acetylcholine concentration that produced 50% of the control maximum response was reduced 17- to 20-fold in bronchus treated with 1 and 10 nmol/L rhIL-5. The lower concentration of 0.1 nmol/L rhIL-5 had no effect. The rhIL-5 effect on bronchial contractility was attenuated by antibodies to IL-5 (TRFK-5; 100 nmol/L) and human IL-5R(alpha) (100 nmol/L). rhIL-5 (10 nmol/L) did not enhance contractility of saphenous vein or jejunum. When RT-PCR was used, IL 5R(alpha) expression was strong in bronchus muscle, weak in trachealis, saphenous vein, and atrial muscle, and undetectable in jejunum, urinary bladder, and pulmonary and renal artery muscle. Comparable weak expression of CCR3 was identified in bronchus and saphenous vein. CONCLUSION: The findings are consistent with an airway tissue-selective expression of the IL-5 receptor that mediates IL-5-induced airway hyperresponsiveness independent of eosinophils. In asthma, in which IL-5 expression is elevated, IL-5 might directly induce bronchial hyperresponsiveness. PMID- 11897985 TI - Rhinitis as an independent risk factor for adult-onset asthma. AB - BACKGROUND: For many years, the association between asthma and rhinitis has primarily been attributed to a common allergic background. Recently, it has been suggested that asthma and rhinitis are associated in the absence of atopy. The nature of this association is not well known. OBJECTIVE: The purpose of this study, which was performed in a large, longitudinal community population, was to determine the extent to which rhinitis is an independent risk factor for adult onset asthma. METHODS: We carried out a nested case-control study from the longitudinal cohort of the Tucson Epidemiologic Study of Obstructive Lung Diseases. One hundred seventy-three incident patients with physician-confirmed asthma were compared with 2177 subjects who reported no asthma or shortness of breath with wheezing. Potential risk factors, including the presence of rhinitis, were assessed before the onset of asthma (patients) or before the last completed survey (control subjects). RESULTS: Rhinitis was a significant risk factor for asthma (crude odds ratio, 4.13; 95% confidence interval, 2.88-5.92). After adjustment for years of follow-up, age, sex, atopic status, smoking status, and presence of chronic obstructive pulmonary disease, the magnitude of the association was reduced but still highly significant (adjusted odds ratio, 3.21; 95% confidence interval, 2.19-4.71). After stratification, rhinitis increased the risk of development of asthma by about 3 times both among atopic and nonatopic patients and by more than 5 times among patients in the highest IgE tertile. Patients with rhinitis with persistent and severe nasal symptoms and a personal history of physician-confirmed sinusitis had an additional increased risk of asthma development. CONCLUSION: We conclude that rhinitis is a significant risk factor for adult-onset asthma in both atopic and nonatopic subjects. The nature of the association between rhinitis and asthma is open to interpretation. PMID- 11897984 TI - Significant variability in response to inhaled corticosteroids for persistent asthma. AB - BACKGROUND: A clinical model is needed to compare inhaled corticosteroids (ICSs) with respect to efficacy. OBJECTIVE: The purpose of this investigation was to compare the relative beneficial and systemic effects in a dose-response relationship for 2 ICSs. METHODS: A 24-week, parallel, open-label, multicenter trial examined the benefit-risk ratio of 2 ICSs in persistent asthma. Benefit was assessed by improvements in FEV(1) and PC(20); risk was assessed by overnight plasma cortisol suppression. Thirty subjects were randomized to either beclomethasone dipropionate (BDP) 168, 672, and 1344 microg/day (n = 15) or fluticasone propionate (FP) 88, 352, and 704 microg/day (n = 15), both administered by means of a metered dose inhaler (MDI) with chlorofluorocarbon propellant via a spacer, in 3 consecutive 6-week intervals; this was followed by 3 weeks of FP dry powder inhaler (DPI) 2000 microg/day. RESULTS: Maximum FEV(1) response occurred with the low dose for FP-MDI and the medium dose for BDP-MDI and was not further increased by treatment with FP-DPI. Near-maximum methacholine PC(20) improvement occurred with the low dose for FP-MDI and the medium dose for BDP-MDI. Both BDP-MDI and FP-MDI caused dose-dependent cortisol suppression. Responsiveness to ICS treatment was found to vary markedly among subjects. Good (>15%) FEV(1) response, in contrast to poor (<5%) response, was found to be associated with high exhaled nitric oxide (median, 17.6 vs 11.1 ppb), high bronchodilator reversibility (25.2% vs 8.8%), and a low FEV(1)/forced vital capacity ratio (0.63 vs 0.73) before treatment. Excellent (>3 doubling dilutions) improvement in PC(20), in contrast to poor (<1 doubling dilution) improvement, was found to be associated with high sputum eosinophil levels (3.4% vs 0.1%) and older age at onset of asthma (age, 20-29 years vs <10 years). CONCLUSIONS: Near maximal FEV(1) and PC(20) effects occurred with low-medium dose for both ICSs in the subjects studied. High-dose ICS therapy did not significantly increase the efficacy measures that were evaluated, but it did increase the systemic effect measure, overnight cortisol secretion. Significant intersubject variability in response occurred with both ICSs. It is possible that higher doses of ICSs are necessary to manage more severe patients or to achieve goals of therapy not evaluated in this study, such as prevention of asthma exacerbations. PMID- 11897986 TI - Early treatment of perennial rhinitis with budesonide or cetirizine and its effect on long-term outcome. AB - BACKGROUND: Perennial rhinitis is a common disease that has many similarities with bronchial asthma. Early treatment with inhaled steroids has improved asthma symptoms, lung function, and bronchial hyperreactivity, but it has not been studied in perennial rhinitis. OBJECTIVE: The main objective was to determine whether early introduction of long-term daily intranasal steroid treatment would have a positive effect on the clinical course and outcome of perennial rhinitis compared with the effect of an antihistamine. A secondary objective was to compare the clinical efficacy of intranasal budesonide and oral cetirizine. METHODS: One hundred forty-three adult patients with newly detected perennial allergic or nonallergic eosinophilic rhinitis of 1 to 3 years' duration were randomized to receive budesonide dry powder, 400 microg (delivered dose of 280 microg) intranasally, or cetirizine, 10 mg orally, once daily for 1 year. At the end of the double-blind treatment period, medication was stopped, and the patients were followed for another year, during which time they could use 14-day courses of intranasal budesonide as needed to control rhinitis relapses. The main outcome measures were the time to first relapse and the number of relapses during the second year. Nasal symptom scores, nasal smear eosinophilia, and nasal peak expiratory flow were used to compare the clinical efficacy of the 2 treatments. RESULTS: During the randomized phase of the study, budesonide was significantly more effective than cetirizine in relieving nasal symptoms. Nasal peak expiratory flow improved significantly in budesonide-treated patients compared with in patients receiving cetirizine. After discontinuation of randomized treatment, 38% of budesonide-treated and 56% of cetirizine-treated patients had a relapse within the first month (P =.04). The median time to first relapse was longer in budesonide-treated patients than in cetirizine-treated patients (62 vs 20 days), although the difference was not significant. Fourteen-day courses of budesonide provided effective control of relapses; the mean number of relapses was 4.0 versus 5.4 in the groups previously treated with budesonide or cetirizine, respectively. Both treatments were well tolerated throughout the study. CONCLUSIONS: Budesonide is significantly more effective than cetirizine in controlling perennial rhinitis. After stopping treatment, budesonide better prevents relapses for 1 to 2 months compared with cetirizine. Periodic therapy with budesonide may be sufficient to control symptoms in most patients who have relapses. PMID- 11897987 TI - Inhaled corticosteroids plus salmeterol or montelukast: effects on resource utilization and costs. AB - BACKGROUND: Experimental clinical studies have demonstrated that the addition of salmeterol to inhaled corticosteroids (ICSs) is superior to the addition of montelukast to ICSs. Observational research from real-world clinical practice is needed to confirm these results. OBJECTIVE: The present study was designed to assess, in clinical practice, the comparative impact on health care utilization and cost of 2 dual-controller therapies, ICS + salmeterol and ICS + montelukast. METHODS: This study involved the use of a 24-month pre/post retrospective design in patients continuously enrolled in any of 14 United HealthCare plans. Outcomes assessed were post-index pharmacy costs, rates of emergency department visits and hospitalizations, numbers of filled prescriptions for short-acting beta-agonists (SABAs), total asthma costs, and total health care costs. RESULTS: Subjects in the ICS + salmeterol group had 35% fewer post-index SABA claims than subjects in the montelukast add-on group (P 0.05). A statistically significant decrease was found in the BT of all the treatment groups compared to the control group (p <0.05). The histopathologic examination of terminal ileum in the control group revealed that the inflammatory infiltration was significantly less than that in the other groups (p <0.05). PMID- 11898028 TI - Do future general surgery residents have adequate exposure to endocrine surgery during medical school? AB - During their general surgical rotations, medical students should ideally have exposure to a wide breadth of surgical procedures, especially if they are interested in pursuing surgical careers. To determine their exposure to endocrine surgery during medical school, we surveyed students from more than 20 medical schools who interviewed for general surgery residency positions at our institution over a 2-year period. Questions focused on the total number of index surgical procedures observed during all of their medical school education. Of 211 surveys sent, 146 were returned (66%). The mean age of the students was 26.0 +/- 0.3 years, and 21% were women. The average times spent on general surgery and surgery subspecialty rotations during medical school were 11.1 +/- 0.6 weeks and 7.6 +/- 0.4 weeks, respectively. The mean number of thyroidectomies (2.8 +/- 0.3), parathyroidectomies (1.9 +/- 0.3), and adrenalectomies (0.5 +/- 0.1) observed by the medical students were significantly lower than operations such as mastectomies (9.4 +/- 0.3), coronary bypass surgeries (8.7 +/- 1.4), and laparoscopic cholecystectomies (10.0 +/- 0.7). Furthermore, of these 146 future surgical residents, 34% failed to observe a single thyroid resection, 42% did not see a parathyroidectomy, and 65% failed to see an adrenalectomy. In conclusion, future general surgery residents seem to observe a wide variety of surgical cases, but most have little or no exposure to endocrine surgery. This paucity of exposure may have significant educational and career ramifications. PMID- 11898029 TI - Pattern of nodal metastasis for primary and reoperative thyroid cancer. AB - This retrospective investigation was undertaken to clarify the pattern of nodal metastasis in papillary (PTC) and medullary (MTC) thyroid carcinoma. Nodal metastases are associated with recurrence of both PTCs and MTCs. The extent of lymph node dissection is controversial owing to the lack of reliable diagnostic criteria for nodal metastases other than histopathology. Between November 1994 and October 1999 a total of 296 patients (134 PTCs, 162 MTCs) underwent total thyroidectomy in conjunction with a standard resection of at least the cervicocentral lymph node compartment. Of 10,446 sampled lymph nodes, 1641 were positive. All nodes were related to their respective cervicomediastinal compartments. The ipsilateral cervicolateral compartment was involved almost as often as the cervicocentral compartment in primary PTC (29% vs. 32%), reoperative PTC (21% vs. 37%), primary MTC (34% vs. 34%), and reoperative MTC (49% vs. 65%). The contralateral cervicolateral and mediastinal compartments were more rarely affected, and were least affected in the primary setting. From these data was derived an individualized surgical strategy for PTC and MTC. This concept rests on the joint resection of cervicocentral and ipsilateral cervicolateral compartments. Depending on tumor entity, surgical status, and primary tumor diameter, additional compartments may have to be cleared. PMID- 11898030 TI - Relation between technetium 99m-methoxyisobutylisonitrile accumulation and multidrug resistance protein in the parathyroid glands. AB - Technetium 99m ((99m)Tc)-methoxyisobutylisonitrile (MIBI) scintigraphy has been reported to be a useful method for visualizing the parathyroid glands for the diagnosis of hyperparathyroidism. Some drug metabolism transporters, such as p glycoprotein (P-GP) or multidrug resistance-associated protein (MRP), are believed to be involved with one mechanism of (99m)Tc-MIBI accumulation in the parathyroid glands. We analyzed the expression of P-GP and MRP in 40 parathyroid glands from eight patients with primary hyperparathyroidism and six patients with secondary hyperparathyroidism using an immunohistochemical procedure. These patients underwent (99m)Tc-MIBI scintigraphy and thallium 201 ((201)Tl)/(99m)Tc subtraction scintigraphy preoperatively. We investigated the relation between the scintigraphic images and the expression of P-GP and MRP. The positive findings of (99m)Tc-MIBI accumulation in the hyperfunctioning parathyroids were increased significantly. There was a significant difference between the expression of P-GP and the (99m)Tc-MIBI scintigraphy findings. High expression of P-GP in parathyroid cells resulted in the negative image of (99m)Tc-MIBI, and low or no expression of P-GP made the image positive. There was no significant difference between the expression of MRP and the (99m)Tc-MIBI scintigraphic imaging. The (99m)Tc-MIBI image was also related to the weight of the parathyroid glands but not to the type of the parathyroid cells. Based on these clinical findings, we can hypothesize that P-GP functions as a drug transporter not only for chemotherapeutic agents but also for (99m)Tc-MIBI. PMID- 11898031 TI - Induction of orthotopic rat adrenomedullary neoplasia by intraadrenal pheochromocytoma cell transplantation. AB - We aimed to establish a technically feasible, easily reproducible model of orthotopic adrenomedullary neoplasia. Male rats received adrenal injection of rat pheochromocytoma cells infected with the Escherichia coli gene for beta galactosidase (lac Z). Each of 10 animals was perfused 7 or 24 days after tumor cell injection; 5 animals of each group were injected with cyclosporin. Animals without tumor cell injection served as controls. Tumor cells were identified and characterized in frozen sections by histochemical and immunohistochemical methods. Immunosuppressed animals had enlarged adrenals 7 days after tumor cell injection. In the rats without immunosuppression the adrenals seemed unaltered despite microscopic demonstration of tumor cells. After 24 days tumors had developed in all animals, weighing 50 times more than normal adrenals in animals with immunosuppression, and 9 times more in animals without immunosuppression. Intraadrenal catecholaminergic tumor cells could be identified by beta galactosidase expression. No animal showed systemic spread. Generation of adrenomedullary neoplasia by intraadrenal pheochromocytoma cell transplantation is easily reproducible and technically feasible. This model allows simultaneous study of neoplastic and normal adrenal tissues (e.g., regarding their response to drugs intended for diagnostic and therapeutic purposes). The decreased tumor growth in animals without immunosuppression is presumably due to the high number of intraadrenal immunocompetent cells. PMID- 11898032 TI - Nonanatomic thoracoscopic wedge resection for diffuse lung disease and indeterminate pulmonary nodule. AB - Video technology has revolutionized thoracoscopy dramatically, considerably increasing its indications. The clinical charts of patients who underwent a video thoracoscopic procedure during a 6-year period were reviewed. Any patient in whom lung wedge resection for diffuse disease or an indeterminate nodule was performed met the inclusion criteria. Early and long-term outcomes were analyzed. A total of 310 thoracoscopic procedures were performed in the 250 patients reviewed. Of these patients, 60 presented with diffuse lung disease and 71 with an indeterminate pulmonary nodule. The total morbidity among diffuse disease patients was 5% (one intercostal vessel hemorrhage and two air leaks). Overall mortality for this group was 11% and was related to previous respiratory status and underlying disease. Patients not requiring preoperative mechanical ventilation ended up requiring it postoperatively, for a crossover rate of 23%. There was no morbidity or mortality in patients who did not require mechanical ventilation. The therapeutic impact index (defined as the total number of patients divided by the patients in whom initiation or withdrawal of specific treatment was due to the biopsy result) for diffuse lung disease was 0.9. Regarding lung nodule resection, early morbidity was present in one patient, who developed a persistent air leak. Late morbidity was present in three patients, who developed persistent intercostal pain. Total morbidity was 5.6%. No mortality was observed for this group. Nonanatomic wedge resection via video-thoracoscopy for diffuse pulmonary disease and indeterminate lung nodule is feasible using minimally invasive methods. Morbidity and mortality are related to the underlying disease and the respiratory status; they are not necessarily due to the procedure. PMID- 11898033 TI - Reduced neutrophil sequestration in lung tissue after laparoscopic lavage in a rat peritonitis model. AB - Laparoscopy to treat abdominal infections is becoming more and more popular. The effects of the CO(2) pneumoperitoneum have not yet been completely clarified. In a rat peritonitis model, therefore, we investigated the influence of laparoscopic lavage in comparison with the conventional technique. A defined multibacterial fecal specimen was installed in the abdominal cavities of 80 rats. These animals were randomized to three groups: group 1 (n = 32), no intervention; group 2 (n = 24), conventional; group 3 (n = 24), laparoscopic lavage. At 1, 2, and 8 hours after the surgical intervention, animals were killed and autopsied. The main outcome measures were bacteremia, interleukin-6 (IL-6) in plasma and ascites, changes in the blood count, and myeloperoxidase (MPO) activity in lung, liver, kidney, and pancreas. Differences of bacteremia were not found. In the ascites a marked increase in IL-6 was observed after 8 hours, which was lower in the treatment groups than in the controls (p <0.025). MPO activity as a measure of the granulocytes present in the tissue showed significant changes only in lung tissue. Two hours after the surgical intervention, the MPO in the lung in the laparoscopy group was significantly lower than that in the controls and the laparotomy group. In conclusion, conventional and laparoscopic lavage reduce inflammation. In this model, laparoscopic lavage with a CO(2) pneumoperitoneum appeared to have no negative influence on the inflammatory reaction during the early postoperative phase. Reduced neutrophil sequestration in lung tissue following laparoscopic lavage reflects the lower level of trauma caused by laparoscopy. PMID- 11898034 TI - New predictor for the defecatory function after sphincter-preserving operation: lumen score. AB - Few studies on sphincter-preserving surgery have analyzed the colon used for the anastomotic segment. We evaluated the usefulness of measuring the square of the diameter of the sigmoid colon (cm(2)) (lumen score, LS) as a predictor of defecatory function after very low anterior resection (VLAR) for rectal cancer. Measurements were done by radiography with semiliquid barium, and the LS was calculated. A total of 24 patients [straight coloanal reconstruction (VLAR-S), n = 17; colonic J pouch reconstruction (LVAR-J), n = 7] were studied more than 6 months after the operation. VLAR-S was divided by the LS results: the high-LS group had an LS of 12 or more (n = 5), and the low-LS group had an LS of less than 12 (n = 12). The neorectal capacity, anal manometry, and defecatory function were studied. In the VLAR-S group, LS had a significant positive correlation with neorectal capacity (gamma = 0.81, p <0.01) and a negative correlation with bowel frequency (gamma = -0.67, p <0.05). Regarding neorectal capacity, the high-LS group had a significantly larger capacity than the low-LS group (118.0 vs. 88.3 ml; p <0.05). The low-LS group had unfavorable defecatory function compared with that of the high-LS group, which was equal to that of the VLAR-J group. We concluded that the LS is a useful predictor of successful colonic J pouch reconstruction. PMID- 11898035 TI - Surgery of colorectal cancer: surgical morbidity and five- and ten-year results in 2400 patients--monoinstitutional experience. AB - The objective of this study was to determine surgical morbidity and long-term outcome of colorectal cancer surgery for quality control reasons and as the basis for new treatment modalities. Surgically treated colorectal cancer patients (mean age 65 years) were followed prospectively in a university center (110 months mean follow-up, 1978-1999). Overall survival (OAS), radicality, extent of resection, recurrence, and morbidity were analyzed (log-rank test of survival, multivariate analysis). Altogether, 2452 colorectal cancers localized in the colon (CC, 44.6%), rectum (RC, 44.8%) or multicentric (CRC, 10.6%) were of UICC stages I (19%), II (30%), III (21%), IV (20%), or undetermined (10%). Radicality and stage but not tumor localization influenced the OAS (p <0.0001). The 5-year/10-year OASs were 50%/42% (all), 78%/66% (R0), 46%/36% (R1), 4%/0% (R2), 0% (unresected) and 86%/79% (I), 70%/58% (II), 42%/33% (III), 3%/0% (IV) or 21%/12% (undetermined), respectively (p <0.0001). Multivisceral resections (17%) resulted in morbidity and survival rates equal to those for standard resection. The overall tumor recurrence rate was 27%, mainly with both local and distant relapse (15%). Surgery-related complications occurred in 18% (all), 14% (CC), 21% (RC), or 20% (CRC). The perineal infection rate (RC) was 4%, overall anastomotic leakage 1%, and mortality rate 0.8%. A prospective, uniform follow-up used over two decades warrants quality control in colorectal cancer surgery, which was curative for half of the patients. The morbidity and mortality were low and were not increased by multivisceral resections. PMID- 11898036 TI - Pectoralis major musculocutaneous flap in head and neck cancer reconstruction. AB - Described initially by Ariyan in the 1970s, the pectoralis major flap has broad acceptance for its versatility in head and neck cancer reconstructions. It is supplied by the thoracoacromial artery, with an additional circulation provided by the lateral thoracic artery. It can be safely used even in patients who have undergone postoperative radiation. The objective of this work is to analyze retrospectively the indications and results of this reconstruction technique in 17 patients with head and neck cancer. We have selected the pectoralis major flap for reconstruction of floor of the mouth and tongue (7 patients); pharyngoesophageal transit after pharyngolaryngectomy (7 patients); facial tissue repair after parotid cancer excision and reconstruction of the soft part of cervical area after skin excision during cervical dissection. Total necrosis of the flap for pharyngoesophageal reconstruction was observed in one patient. Partial loss of the skin flap with partial dehiscence occurred in four patients underwent tongue and floor of the mouth repair, but without fistulae and infection. There was donor site seroma in one patient. The flap was functionally adequate both in the reconstruction of the neopharynx and for repair of great surgical defects. So it is a versatile method with good functional results. PMID- 11898037 TI - Squamous cell carcinoma of the esophagus: multimodal therapy in locally advanced disease. AB - The aim of this prospective study is to report our experience in the multimodal management of locally advanced esophageal squamous cell carcinoma (LAESC; stage III cTNM), focusing on the results of chemoradiotherapy followed by surgery. These findings were compared to the results of a standard group of patients with locally advanced esophageal carcinoma (LAEC; stage III pTNM) treated in our center with surgery alone. Sixty-one patients with LAESC underwent preoperative chemoradiotherapy (5-fluorouracil + cisplatin) with concomitant 45 Gray radiotherapy in a 5-week course. Transthoracic esophagectomy was performed 4 to 5 weeks after the end of the neoadjuvant therapy. Thirty-eight patients underwent surgery, and 37 of them had resections (resectability: 97% in the multimodal group; 84% in the standard surgical series; p = 0.07). The R0 (complete) resection rate was 78% compared to 56% in the standard surgical group (p <0.03). Eleven patients had no residual tumor in the resected specimen (pathologic complete response: pCR: 30%). The operative mortality rate was 19% compared with 8.8% in the standard series. The overall median survival of the resected patients was 21 months, with a 5-year survival rate of 11% (14% in the surgical group; NS). The 3-year and 5-year survival rates were 34% for the pCR group and respectively 5% and 0% for the group with pathologic incomplete response (pIR; p <0.05). The median survival was 28 months for the pCR patients and 19 months for the pIR group. In this non-randomized trial, preoperative chemoradiotherapy in LAESC seems to increase the resectability and R0 resection rates, to allow a higher pCR rate and a longer survival only in the pCR group, at the expense of an inadequate increase in operative mortality. This multimodal treatment cannot be proposed as a standard procedure unless less toxic regimens are developed, increasing the benefits with better local and distant failure control and decreasing operative mortality. PMID- 11898038 TI - Conversion of laparoscopic cholecystectomy to open cholecystectomy in acute cholecystitis: artificial neural networks improve the prediction of conversion. AB - Laparoscopic cholecystectomy is now also performed for acute cholecystitis. In the presence of inflammatory conditions, technical difficulties leading to conversion to open cholecystectomy may occur and overshadow the advantages of the laparoscopic approach. Factors associated with these undue events combined with techniques capable of learning from them may help in determining when to completely avoid the laparoscopic procedure. In this study we determined predictors of conversion in acute cholecystitis and tested their predictive ability by means of statistical multivariate analysis and artificial neural networks. Between January 1994 and February 1997, 225 patients underwent laparoscopic cholecystectomy for acute cholecystitis. Preoperative and operative data were prospectively collected on standardized forms. The first 180 laparoscopically approached cases entered the training set, which was learned by both the statistical and the artificial neural networks methods. Conversion was first studied in relation to a set of preoperative data. Prediction models were then fitted by both of these methods. The last 45 operated cases, which remained unknown to the learning systems, served for testing the fitted models. The forward stepwise logistic regression technique, the forward stepwise linear discriminant analysis, and the artificial neural networks method enabled positive prediction of conversion in 0%, 27%, and 100% of the cases, and a negative prediction in 80%, 85.5%, and 97% respectively, in the training set. A positive prediction of conversion in 0%, 25%, and 67% of the cases, and a negative prediction in 82%, 88%, and 94%, respectively, in the untrained, validation set of patients. An artificial neural networks based model provides a practical tool for the prediction of successful laparoscopic cholecystectomies and their conversion. The high degree of certainty of prediction in untrained cases reveals its potential, and justifies, under appropriate conditions, the complete avoidance of laparoscopy and turning directly to open cholecystectomy. PMID- 11898039 TI - Treatment of gallstone pancreatitis: six-year experience in a single center. AB - Acute pancreatitis (AP) is a complicated disease in 20% to 25% of cases and carries a mortality of 8% to 15%. Etiologically, the most frequent form is acute biliary pancreatitis. Treatment of such an entity is still controversial, but minimally invasive techniques undoubtedly play an important role. We retrospectively analyze our cases of AP observed from January 1992 to May 1998. Etiology was biliary in 95/125 (76%) cases. In 90 cases we evaluated the patient within a few hours of the onset of symptoms. According to the Ranson criteria, we observed a mild form in 74/90 (82.2%) cases and a severe form in 16/90 (17.8%) cases. Our standard policy was to perform urgent endoscopic retrograde cholangiopancreatography (ERCP) with endoscopic sphincterotomy followed by elective laparoscopic cholecystectomy. In particular clinical settings, other modalities of treatment were employed, such as percutaneous cholecystostomy and percutaneous drainage of fluid collections. Successful ERCP was performed in 86/90 cases (95.5%). The procedure was performed in an emergency setting (within 48 hours) in 62/90 cases (68.9%). Whenever the patient was a candidate for surgery, cholecystectomy was performed, laparoscopically in 67/90 cases (74.4%) and via laparotomy in 7/90 cases (7.8%). In only two cases was pancreatic necrosectomy necessary. Globally, we observed a low procedure-related morbidity (6.7%) and no mortality. In the setting of acute biliary pancreatitis, regardless of the severity of the attack, an urgent ERCP + endoscopic sphincterotomy followed by laparoscopic cholecystectomy is safe and could enable successful management of the patient. Associated morbidity and mortality were low. In addition, when indicated, it is possible to treat a great number of concomitant complications with percutaneous ultrasound-guided drainage. PMID- 11898040 TI - Clinical and genetic analysis of noncancerous and cancerous biliary epithelium in patients with pancreaticobiliary maljunction. AB - We analyzed clinical features and genetic alterations in the noncancerous and cancerous biliary lesions obtained from pancreaticobiliary maljunction (PBM) patients. Gallbladder (GB) and bile duct (BD) lesions were obtained surgically from 36 patients with PBM, and polymerase chain reaction (PCR) methods were used to examine for mutations of the K-ras gene and the p53 gene and for microsatellite instability (MSI). The 36 cases were clinically classified into two types according to whether extrahepatic bile duct dilatation was present: a congenital choledochal dilatation (CCD) group (n = 20) and a noncongenital choledochal dilatation (NCCD) group (n = 16). In the NCCD group, all 16 GB specimens exhibited hyperplastic, dysplastic, and cancerous (n = 9) lesions, but no pathological lesions were detected in the 12 BD specimens. On the other hand, in the CCD group, pathological examination revealed lesions, including 8 cancerous lesions, in 60% of the 20 GB specimens and lesions, and including 8 cancerous lesions, in 65% of the 20 BD specimens. K-ras mutations and MSI were detected in 33.3% and 0%, respectively, of 9 hyperplastic lesions, 28.6% and 85.7%, respectively, of 7 dysplastic lesions, and 60.0% and 80.0%, respectively, of 25 cancerous lesions (p <0.05; MSI in hyperplasia vs. dysplasia and cancer). There was no difference of the frequency in K-ras mutations and MSI between the NCCD and CCD groups. By contrast, p53 mutations were detected only in the cancerous GB lesions of both types, the rate being 35.3%. Genetic alterations of K-ras, MSI, and p53 are strongly associated with biliary tract cancer in PBM patients. MSI appears to contribute to carcinogenesis in the biliary tract mucosa of PBM patients, and p53 mutations may be related to the development of GB cancer in the CCD group. PMID- 11898041 TI - Duct-to-mucosa pancreaticojejunostomy reduces the risk of pancreatic leakage after pancreatoduodenectomy. AB - The aim of this retrospective study was to analyze the risk factors for pancreatic anastomotic leakage after pancreatoduodenectomy (PD) and to determine whether duct-to-mucosa pancreaticojejunostomy is superior to the total external tube drainage technique. Between 1990 and 1999, 161 patients underwent PD with end-to-side pancreaticojejunostomy at our institution. Fourteen preoperative and ten intraoperative risk factors for pancreaticojejunal anastomotic leakage were analyzed. Pancreaticojejunal anastomotic leakage was identified in 11% (17/161) of the patients. No preoperative parameters were found to have a significant association with the risk of pancreatic leakage. Three intraoperative parameters were identified as significant by means of univariate analysis: anastomotic technique, pancreatic duct size and texture of the remnant pancreas. A duct-to mucosa pancreaticojejunostomy with total external tube drainage (3% vs. 15%, p = 0.018). A pancreas without duct dilatation of soft pancreas was more likely to develop pancreatic leakage than one with duct dilatation or atrophy. A multivariate analysis revealed that only anastomotic technique turned out to be an independent risk factor (Odds ratio: 4.15, CI: 1.1-27.4). Sub-analysis of patients with soft pancreas and non-dilated pancreatic duct further supported the finding that the duct-to-mucosa pancreaticojejunostomy technique is safer for patients at high risk. Results indicate that the status of the remnant pancreas and the pancreaticojejunostomy technique are the substantial risk factors for pancreatic leakage after pancreatoduodenecomy. Duct-to-mucosa pancreaticojejunostomy might well be the procedure of choice. PMID- 11898042 TI - Preoperative portal embolization in patients with hepatocellular carcinoma. AB - The factors that contribute to the effect of portal vein embolization before hepatectomy for hepatocellular carcinoma are unclear. Sixty-six patients with hepatocellular carcinoma were enrolled in the study. Changes in liver function, portal vein pressure, and liver volume after embolization were examined. A multiple linear regression analysis was performed to identify factors that independently contributed to the effects of portal vein embolization. The acceptable volume ratio of the remnant liver was calculated from liver function and compared with the volume ratio of the non-embolized liver. No postoperative deaths were observed after portal vein embolization or hepatectomy. Serum total bilirubin and prothrombin time did not change significantly after portal vein embolization. In patients who underwent arterial embolization before portal vein embolization, aminotransferase levels increased significantly. The only factor that could significantly predict the atrophy effects of portal vein embolization was previous arterial embolization. The volume ratio of the non-embolized liver was smaller than the acceptable volume ratio of the remnant liver in 18 of 40 patients and increased over the acceptable volume ratio in all cases after portal vein embolization. Portal vein embolization induced atrophy or hypertrophy of the embolized or non-embolized liver sufficiently, even when the liver was dysfunctional or cirrhotic. The atrophy effects were significant, especially when arterial embolization had been performed before portal vein embolization. PMID- 11898043 TI - Laparoscopic splenectomy for idiopathic thrombocytopenic purpura: outcome and long-term results. AB - The technical feasibility of laparoscopic splenectomy (LS) has been recently established. However, data regarding the efficacy of the procedure with long-term follow-up of patients with idiopathic thrombocytopenic purpura (ITP) are scarce. The objective of this study was to determine retrospectively the immediate efficacy and the long-term results of a standardized laparoscopic procedure applied to patients with ITP refractory to medical treatment. Laparoscopic splenectomy was performed in 35 patients for ITP between May 1993 and May 1998. The lateral approach was used in the last 27 patients. Data were recorded retrospectively on that group. Twenty-six patients (96%) underwent successful LS. Mean operative time for the laparoscopic procedure was 90 minutes. There were no postoperative deaths. Postoperative complications developed in three patients. Thrombocytopenia resolved after surgery in 93% of patients, but 7 patients (25%) experienced relapse during a mean 28-month follow-up. At present no patient needs medical therapy to maintain a normal platelet count. Laparoscopic splenectomy is feasible and safe in patients with ITP. Long-term results of LS for ITP are comparable to those achieved with open splenectomy. PMID- 11898044 TI - Lower limb ischemia-reperfusion injury triggers a systemic inflammatory response and multiple organ dysfunction. AB - Restoration of blood flow to an acutely ischemic lower limb may, paradoxically, result in systemic complications and unexpected mortality. We investigated the effect of acute ischemia-perfusion of the lower limb on cytokine production and end organ function. Plasma concentrations of tumor necrosis factor-alpha (TNF-a) and interleukin-6 (IL-6) were determined in five groups of male Wistar rats: control, 3 hours of bilateral hind limb ischemia alone, and 3 hours of bilateral hind limb ischemia followed by 1 hour, 2 hours, or 3 hours of reperfusion, respectively. In a second experiment, the effect of lower limb ischemia reperfusion on remote organs (lung, liver, and kidney) was assessed biochemically and histologically. There was a significant increase in plasma concentrations of TNF-a in plasma of animals subjected to 3 hours of bilateral hind limb ischemia followed by 1 hour of reperfusion, 40.1 +/- 4.4 pg/ml, when compared with controls, 22.6 +/- 4.4 pg/ml, or animals in the ischemia-alone group, 16.3 +/- 5.2 (p <0.05). Plasma concentration of IL-6 increased progressively and significantly in animals subjected to bilateral hind limb ischemia followed by 1 hour of reperfusion, 720 +/- 107 pg/ml; 2 hours of reperfusion, 1987 +/- 489 pg/ml; or 3 hours of reperfusion, 6284 +/- 1244 (p <0.0001), compared with controls, 104 +/- 43 pg/ml; or animals in the ischemia-alone group, 140 +/- 55 pg/ml. In the study comparing portal and systemic concentrations of IL-6, systemic concentrations of IL-6, 967 +/- 184 pg/ml were significantly higher than those in the portal circulation 577 +/- 127 pg/ml (p <0.05). There was a significant increase in plasma concentrations of urea, creatinine, aspartate transaminase, alanine transaminase, and lactic dehydrogenase in reperfused animals compared with controls (p <0.001). Morbidity and mortality following reperfusion of the acutely ischemic limb may be a manifestation of multiple organ dysfunction caused by a systemic inflammatory response triggered by reperfusion of the ischemic extremities. PMID- 11898045 TI - Surgical implications in the current treatment of Wolff-Parkinson-White syndrome. AB - There are currently two different surgical approaches to the abnormal pathway, Wolff-Parkinson-White (WPW) syndrome-the endocardial (ENDO) and epicardial (EPI) techniques. In recent years, ablation of accessory pathways can be achieved by catheter-induced radiofrequency (RF) current. This study was undertaken to assess our results of surgical treatment for WPW syndrome in the current era of catheter ablation. From 1985 to 1993, 51 patients (33 male and 18 female) with WPW syndrome underwent operations for ablation of accessory pathways. Associated anomalies included Ebstein's anomaly, coronary artery disease, and tricuspid atresia. Preoperatively, 6 patients underwent unsuccessful RF catheter ablation. Fifteen (29%) patients were operated with the ENDO technique and 36 (71%) with the EPI technique. There was no early death in either group. In the immediate postoperative period 40 (78%) patients were in sinus rhythm. The electrophysiological studies revealed successful ablation of the pathway in 50 (98%) patients. On complete late follow-up (mean, 36 months) all patients were back to preoperative levels of activity. Our experience indicates that excellent results can be achieved with each of these two techniques. The left free wall accessory pathways may be ablated in a more reproducible way with the ENDO approach. The concept that surgical ablation of accessory pathways may prevent further atrial fibrillation is supported by the low incidence in this series. Operations for WPW syndrome may become indicated for RF ablation failure, when additional procedures are required. In these cases the surgical skill should be available, and this is a skill that should not be lost. PMID- 11898046 TI - Angelchik prosthesis revisited. AB - There are few long-term follow-up reports of the Angelchik prosthesis (AP). We report the longest follow-up series (66-192 months, average 145 months) to date. Between October 1983 and January 1994, 65 patients (45 men and 20 women) aged between 29 and 84 years (mean 52 years) had an AP inserted for gastro-oesophageal reflux (GOR) with or without hiatus hernia (HH). Clinical, radiological, endoscopy, and operative details were reviewed. Postoperative complications, investigations, and follow-up details were critically analyzed. All living patients (n = 53) with an AP in situ were interviewed and symptomatic assessment was carried out using a modified Visick system (I-IV). The average duration of the GOR symptoms before the operation was 5.7 years (range 10 months to 20 years). The average hospital stay was 8 days (range 5-15 days). Postoperatively, five patients developed chest infection/atelectasis, four had superficial wound infection, two had deep vein thrombosis (one with pulmonary embolism), one had urinary retention, and four developed an incisional hernia. Six patients (three with an AP in situ) died of other medical conditions. Ten (15%) patients had removal of the prosthesis. Eight (12%) and 11 (17%) had transient and persistent dysphagia, respectively. Thirteen (20%) and five (8%) patients had distal slippage and proximal migration of the prosthesis, respectively. One patient had erosion of the AP into the stomach, while in another patient, the straps of the prosthesis ruptured. Of the 53 living patients with an AP in situ, 28 (53%) were Visick I, 11 (20%) were Visick II, 11 (20%) were Visick III, and 3 (7%) were Visick IV. We conclude that the AP has poor long-term results, with only 66% attaining Visick I and II, and a prosthesis removal rate of 15% (10/65). Patients with preoperative dysphagia, hypothyroidism, and diabetes tend to do worse with an AP. Obese patients and those with failed previous fundoplication seemed to fare well with an AP. In view of poor long-term results and high incidence of complications as compared to other conventional operations for GOR, we cannot recommend the continued use of the AP. PMID- 11898050 TI - [Methods, indications and validation of intraoperative nerve conductivity testing]. AB - During nerve surgery, electrodiagnostic methods are applied to assess the availability and viability of nerve fibers and to adjust operative measures accordingly. The validity of this procedure is verified by histology and by the outcome of the operation. This paper explains the techniques of intraoperative nerve action potential (NAP) and somatosensory evoked potential (SEP) recording, how to interpret the electrodiagnostic results, and describes both the special features and the limitations of the methods. We found reliable results of neurography, detecting the presence or absence of spontaneous nerve regeneration across a lesion in continuity months before the reinnervation reached its final target. Based on our results, we suggest that NAP recording of the exposed nerve can widely prevent unnecessary nerve or fascicle resection. Besides this important indication, the nerve function evaluation was successfully used in nerve surgery whenever the quality of the nerve parenchyma was crucial to the operative management. Further indications such as evaluating brachial plexus lesions and the condition of nerve roots, judging the proximal coaptation site in nerve reconstruction, tracing the site of a nerve lesion and identifying the pathophysiology of nerve malfunction are exemplified. Intraoperative nerve conductivity testing should not be considered as a replacement of but rather as a complement to preoperative clinical, electrophysiological and imaging evaluations and a thorough intraoperative morphological examination. PMID- 11898051 TI - [Lipoma as a rare cause of nerve compression syndrome in the hand and forearm]. AB - We report on three patients who were operated on peripheral nerve compression syndromes in the hand and forearm caused by a lipoma. After surgical removal of the lipoma, distinct improvement or even recovery of symptoms was observed. PMID- 11898052 TI - [Treatment of bites to the hand and wrist--is the primary antibiotic prophylaxis necessary?]. AB - Animal bites make up a large proportion of the injuries treated in an emergency department. Due to the type of injury, the variety of wounds and the contamination with aerobic and anaerobic organisms, they deserve special attention. In this study, we reviewed 98 patients (55 male/43 female) with bite wounds to the hand and wrist treated between 1995 and 2000. They were either treated conservatively (n = 65) or surgically (n = 33) depending on the clinical findings. In 18 of 33 cases, the reason for surgical treatment was an infection. A primary antibiotic prophylaxis, usually with cephalosporines, was administered in 47 of 98 cases. Results were analysed retrospectively. An infection developed in 32 patients. In six of these patients, an infection developed despite primary antibiotic prophylaxis. Operative treatment became necessary in four of these six cases. Twenty-six of 32 patients were treated without primary antibiotic prophylaxis. Surgical treatment was required in around half (n = 14) of these patients, while the other 12 patients were treated conservatively with antibiotic therapy.Twenty-one of 26 patients presented with bite wounds that were already infected. Microbiological examination revealed a variety of microbes, usually a mixed infection with Pasteurella multocida was found. All organisms were susceptible to treatment with second or third generation cephalosporines.A total of 15 patients had to be operated due to deeper injuries to the bone, extensive soft-tissue injury, or because of injury to a tendon and the tendon sheath. In most patients, a good to acceptable functional result was achieved. Primary antibiotic prophylaxis does not prevent the development of infection. Nevertheless, because of the inherently high infection risk associated with bite wounds to the hand and wrist, prophylaxis should be carried out. In case of severe damage to the soft tissue or signs of infection, early surgical therapy should be considered. PMID- 11898053 TI - [Functional outcome and quality of life after ray amputation versus amputation through the proximal phalanx of the index finger]. AB - Little data is available about the long-term functional outcome and quality of life after ray amputation or amputation at the level of the proximal phalanx of the index finger. The purpose of this study was to evaluate the functional outcome and postoperative quality of life after ray amputation or amputation through the proximal phalanx to create a database which is helpful in the decision whether to amputate a digital ray or to preserve a stump.58 patients with amputation of the second ray and 12 patients with amputation through the proximal phalanx of the index finger between 1987 and 1996 were included in the study and examined with respect to hand strength, sensibility, range of motion, pain, and aesthetic result of the hand. Subjective functional outcome was evaluated using the DASH-questionnaire. The majority of patients were male (78 % with ray amputation/83 % with amputation through the proximal phalanx). In 55 %/58 % the operation was performed on the dominant hand. Average age was 45 years and average follow-up was 44.2 months after ray resection and 22.1 months after amputation through the proximal phalanx. Patients lost an average of 10.7 weeks of work after ray amputation and 8.1 weeks after digital amputation. There was no significant loss of grip strength after ray amputation (29 % to 34 % loss of grip strength, 32 % loss of pinch grip) compared with patients after amputation through the proximal phalanx (21 % to 28 % loss of grip strength, 17 % to 35 % loss of pinch grip). DASH-score was 31.3 after ray amputation and 21.7 after digital amputation. Patients with amputation through the proximal phalanx reached a significantly better result in part B of the DASH-questionnaire. 65.5 % of the patients after ray amputation and 91.7 % after digital amputation complained of postoperative pain in the operated hand. Decreased sensibility was found in 55.2 % after ray resection and in 33.3 % after digital amputation. All patients after amputation through the proximal phalanx but only 82.8 % after ray amputation showed a free range of motion of the operated hand. The aesthetic appearance of the operated hand was rated higher after ray amputation. The results show that there is no significant loss of strength after ray amputation compared to amputation through the proximal phalanx as mentioned in the literature. Patients with amputation through the proximal phalanx demonstrate a better functional outcome, while the aesthetic appearance was rated higher after ray amputation. A significant difference was only found in part B of the DASH-questionnaire. This should be considered when the indication for ray amputation is pending. PMID- 11898054 TI - Operative treatment of the chronic post-traumatic boutonniere deformity by the technique of pieper. AB - We report eleven cases of chronic, non-fixed, post-traumatic boutonniere deformity treated by the technique of Pieper: One lateral slip was mobilised and distally desinserted. At the PIP, the slip was pulled through the central slip and sutured in correct tension to the released opposite lateral slip. 73 % regained full extension at the proximal interphalangeal joint. Despite a mild extension deficit of 15 degrees at the distal interphalangeal joint in 64 %, ten of eleven patients were content with the post-operative outcome. We conclude that the technique of Pieper is a good and valuable operative procedure to correct chronic non-fixated button-hole deformity. PMID- 11898055 TI - [Closed reduction and percutaneous K-wire fixation of Bennett's fracture dislocation]. PMID- 11898056 TI - [The Epping resection-suspension arthroplasty procedure. A standard procedure in the operative treatment of trapeziometacarpal osteoarthrosis?]. AB - The surgical treatment of painful osteoarthrosis of the trapeziometacarpal joint with the Epping technique consists of excision of the trapezium and reconstruction of the first intermetacarpal ligament by using the distally based half of the flexor carpi radialis tendon. This ligament reconstruction procedure aims to prevent proximal migration of the first metacarpal, restore function and stability of the neoarthros. This retrospective study presents operative outcome results after using the Epping arthroplasty technique. The Epping technique has been performed in seventy cases and could be evaluated after a mean follow-up of 34.6 months in 92.4 % of all operated patients. Subjective results were evaluated using patient-based questionnaires such as the Buck-Gramcko-Score and the German Version of the DASH V2.0 questionnaire. Objective and functional outcome analysis including range of motion, strength measurements with the computer-based JAMAR dynamometer and various X-ray views, have been used as further methods of evaluation. Excellent pain relief and very good subjective results with 86 % patient satisfaction have been reported by our patients. The objective outcome analysis demonstrated good functional results with respect to radial abduction (51 degrees) and palmar flexion (45 degrees), and improvement in grip strength, key pinch and pulp pinch. Some patients reported remaining problems with pain during performance of activities of daily life and professional activities. A significant proximal metacarpal migration without correlation to the objective or subjective outcome was found in follow-up X-ray controls. The Epping ligament reconstruction procedure has proven to be a valuable standard procedure after a mid-term follow-up period. Good functional outcome and high patient satisfaction could be achieved, but some patients still remain with problems in different activities. Long-term results still need to be evaluated. PMID- 11898057 TI - [Digital nerve block anaesthesia: historical development and two cases of finger tip necrosis, a rare complication]. AB - Two cases of finger-tip necrosis following digital blocks are presented. These are rare complications of this technique. Pathogenesis and treatment options are discussed and the literature reviewed. Circulatory problems can be avoided by using adrenalin free anaesthetics, infiltrating at the metacarpal level with small volumes, and using upper-arm tourniquets instead of rubber bands at the phalangeal level. PMID- 11898058 TI - [Vascularisation of a free jejunal graft at the neck in an insufficient vascular situation]. AB - Free jejunal grafts are the method of choice for one-stage reconstruction of the hypopharynx and the upper esophagus. Prerequisites for a successful free transplantation are a sufficient arterial vascularisation and a corresponding venous drainage. In a 59-year old patient, an esophagectomy was performed because of a proximal esophageal cancer. The passage was primarily reconstructed by a pedicled colon interposition. Necrosis of the cranial graft occurred. After resection of the necrotic bowel, a free jejunal graft was introduced thoraco cervically. The nutrient vessels of the graft were anastomosed to the upper thyroid artery and the vena thyroid ima. In an operative revision one day postoperatively due to lacking re-capillarisation, the arterial inflow stopped. The superior thyroid artery was cut at its cranial origin at the external carotid artery and microsurgically implanted into the common carotid artery. Hereafter, a venous outflow was re-established. But the critical time of ischaemia was exceeded. In another operative revision, the cephalic vein was exposed at the cranial upper arm, distally cut and anastomosed to the re-implanted superior thyroid artery. After the harvest of a second autogenous jejunal graft, the arterio-venous shunt was cut and the segments were anastomosed to the mesenterial artery and vein. Transpositions of arteries and veins with temporary formation of an arterio-venous shunt may be prerequisites for a sufficient vascularisation and can enable a successful free microsurgical transplantation even in critical vascular situations. PMID- 11898059 TI - [The journal for orthopedics - consistency and changes]. PMID- 11898060 TI - [Influencing factors of the patella alignment in TKR and operative soft tissue management]. AB - The patella has a crucial effect on the postoperative result after TKR, as complication statistics demonstrate. What factors influence the patella alignment and through which surgical steps can it be centred? METHOD: Reviewing the findings from publications ranging from 1983 to 2000 and our own operative experience, we tried to establish all factors that could influence patella alignment after TKR. RESULTS: A complex overview is given about the dependency of the patella alignment on the design of the prosthesis, the soft tissue and bone preparation and the implantation technique, together with suggestions for intraoperative management for patella centralisation. CONCLUSION: To avoid any patella pathologies, it is of crucial importance to have comprehensive knowledge and to follow the described surgical steps strictly for patella centralisation, including the extensor mechanism. This is essential to achieve a satisfying functional result after TKR. PMID- 11898061 TI - [Determination of the axis after totalendoprosthesis of the knee: functional X ray photography as golden standard]. AB - AIM: To examine the question of which X-ray photography technique should be given preference after implantation of total endoprosthesis of the knee. METHOD: 20 patients (age 74.1; 12 men, 8 women) with total endoprosthesis of the knee (Type Sigma, PFC, Johnson and Johnson) because of primary athrosis. Postoperatively X ray photographs of the whole leg in standing and 40 x 20 a. p. in a flat position were taken. The angles of the axis were determined and the information analysed. RESULTS: By means of X-ray photography of the whole leg we obtained the following angles: caput-collum-diaphyse 125 +/- 4.3 degrees, femur/basis of the knee 83.2 +/- 2.6 degrees, mechanical femur axis/basis of the knee 89.3 +/- 1.8 degrees, tibia/basis of the talus 89.2 +/- 2.2 degrees. With the 40 x 20 a. p. X-ray photographs we determined the following angles: femur/basis of the knee 82.3 +/- 3.1 degrees, basis of knee/tibia 92.2 +/- 3,6 degrees. Correlating the angles of femur/basis of the knee and basis of the knee/tibia were relations of 0.42 (p < 0.05) and 0.27 (p < 0.05). CONCLUSION: For gaining information about the axis of the leg under functional conditions, one should give preference to X-ray photography of the whole leg. PMID- 11898062 TI - [Outcome of inpatient rehabilitation following total knee replacement using the HSS-Score]. AB - AIMS: The aim of the study was to evaluate the outcome of inpatient rehabilitation following total knee arthroplasty using the HSS score (Hospital for Special Surgery). Factors influencing the result were analysed. METHOD: 182 patients admitted to a rehabilitation unit after total knee replacement were included. There were 146 female and 36 male patients, with mean age of 72 (+/- 6.1) years (range; 49 to 84 years). RESULTS: At admission, the HSS score was 47.4 (+/- 10.6) and was significantly improved to 69.8 (+/- 8.8) at discharge. The percentage of patients with a poor result was markedly decreased from 86.8 % to 10.4 %. The outcome was independent of sex, age, concomitant diseases or postoperative weight bearing. 34 patients could not be discharged after 3 weeks as usually and stayed for another 8.3 (+/- 2.6) days. These patients had significantly lower scores at admission (41.9 +/- 11.2) than patients who stayed for three weeks (48.6+/- 10.1). At discharge, scores were comparable with 67.7 (+/- 9.4) and 70.2 (+/- 8.6), respectively. CONCLUSION: Inpatient rehabilitation following knee arthroplasty significantly improved the HSS score independent of sex, age, concomitant diseases and weight bearing. PMID- 11898063 TI - [Dose-dependent prevention of early periprosthetic bone loss by alendronate]. AB - AIM: Periprosthetic bone loss occurs in the first six months after total hip arthroplasty (THA) and is felt to be largely the result of initial operative irritation, immobilization, and stress shielding. This study (a prospective, randomized, open, blinded endpoint evaluation) aims at preventing bone loss around the stem with an oral bisphosphonate. METHOD: 66 healthy subjects with uncemented THA and low lumbar bone mass density (BMD) (negative T score) were treated post-operatively with alendronate as follows: n = 21 with 10 mg/d for 10 weeks (A), n = 21 20 mg/d for 5 weeks (B), n = 24 no treatment for controls (C). The periprosthetic BMD in the Gruen zones (ROI) was measured after the 2nd, 4th, 6th, and 12th month by DEXA as a percentage of the value measured one week after surgery. RESULTS: In C, there was significant bone loss in all ROI during the first months and a deficit of 29 % in ROI 7 following one year. In B, bone loss was completely prevented up to the second month, in ROI 7, a significant difference in comparison to C was registered for the entire year. In A, significant bone loss reduction during 12 months was seen. CONCLUSION: Alendronate, therefore, is capable of preventing initial periprosthetic bone loss. A dosage of 20 mg/d is required initially with daily treatment lasting at least 10 weeks. PMID- 11898064 TI - [Patellofemoral pain syndrome in young men compared to a normal population exposed to the same physical strain]. AB - AIM: The causes of the patellofemoral pain syndrome, especially in young patients, continue to be largely unknown. The purpose of the study was to determine the extent to which weight and activities that impose a strain on the knee influence the occurrence of patellofemoral pain in young, physically active men. METHOD: 248 soldiers on mandatory military service were included in the study. 133 persons had pain in the knee prior to or during military service while 115 persons had no symptoms in the knee. Anthropometric data and patient history were recorded for all individuals. This was followed by clinical examination. RESULTS: Age distribution and body mass index were identical in both groups. However, significantly more subjects in the group with pain prior to military service had been active sportsmen while a significantly larger number of normal individuals had had jobs that strained the knees. The greatest difference, however, was seen in the number of previous injuries. These were much more common in the group that had pain and the injuries most commonly were contusions. CONCLUSION: It is well known that previous injuries and sports favor the emergence of patellar pain. However, persistent strain on the knee at work appears to be less commonly associated with this symptom. The individual's body mass index was of no importance in this context. PMID- 11898065 TI - [MR-Arthrography with radial sequences for visualization of early hip pathology not visible on plain radiographs]. AB - AIM: We wanted to improve MRI visualization of early hip pathology invisible on plain radiographs. METHOD: An MR arthrography of the hip with radial sequences is described. The standard MR technique is improved by 1) using a small flexible surface coil to show selectively the hip joint to be examined, 2) application of gadolinium intra-articularly and 3) by radial imaging sequences perpendicular to the true plane of the acetabulum. CONCLUSION: By this technique it is possible to achieve an undistorted image of each aspect of the acetabular rim. Regions of special interest can be defined and their pathologies possibly related to morphologies of the proximal end of the femur. PMID- 11898066 TI - [Osteochondritis dissecans - an easy classification in MRI]. AB - AIM: For the diagnosis of osteochondritis dissecans (OD) MRI is used besides X ray. However, there is no consensus about the necessity of MRI or the interpretation of MRI. The aim of this prospective study was to find a diagnostic concept for OD which is simple and practicale for daily routine. METHOD: In 90 patients with OD of the knee or ankle MRI (T1- and T2-w SE) was carried out before arthroscopy. According to the interface between the osteochondral fragment and the parent bone, MRI was classified in 2 stages ("stable" or "unstable") and compared with the arthroscopic findings. RESULTS: With the 2-stage classification of MRI, a prediction of the stability of the osteochondral fragment was possible in 92 %. Out of the 90 patients 7 (8 %) differed regarding preoperative stability in MRI compared to stability in arthroscopy. Incorrect diagnosis in MRI was only found among the patients with unstable fragments. CONCLUSION: The chosen 2-stage classification in MRI is well suitable for the planning of stage-related therapy. PMID- 11898067 TI - [Neuromuscular failure of the rotator cuff as a contribution to the functional impingement of the shoulder - a muscle biopsy investigation]. AB - AIM: The possibility that functional impingement results from muscular imbalance is under discussion. This study investigates whether the disturbance of muscular coordination could be demonstrated as a shift in the balance of FT and ST fibers, since the FT fibers enable a rapid reaction or contraction, and ST fibers enable slow contractions and sustained performance. METHOD: In 37 patients with a supraspinatus syndrome, partial or complete rupture of the rotator cuff, we took biopsies from the supraspinatus and deltoid muscles and did morphometric measurements.< RESULTS: All three groups of patients showed a reduction in size, increased variability and change in frequency of the fiber cross sections. The FT fibers are especially affected. These findings are objectively confirmed by morphometric measurements. CONCLUSION: A disturbance in coordination of the musculature which can bring about functional impingement might be inferred from the above changes. We term this "neuromuscular insufficiency of the rotator cuff". The mechanical factors relating to subacromial impingement probably only have a role in promoting its manifestation by restricting the compensation range of the joint. PMID- 11898068 TI - [Reliability of active range-of-motion measurement of the rotation in the forearm: comparison of three measurement devices]. AB - AIM: The aim of this study is to prove the intra- and interobserver reliability of three different methods to measure the active range-of-motion in the forearm. METHODS: These three different methods were tested on forty volunteers. In the first method pictures were made in full supination and pronation, measurement of the two angles was performed with a dedicated software. Active range-of-motion was defined as the magnitude of forearm rotation between maximum pronation and supination. The second method used a standard goniometer. The third method of assessment used a gravity goniometer (plurimeter). We randomised both the side of the patient to be examined and the sequence of the methods. Two examiners measured subsequently the range-of-motion twice with these three devices. RESULTS: We found a significant intraobserver reproducibility with the gravity goniometer. There was no significance with the goniometer or pictures. The intraobserver reproducibility of the latter two methods was poor. The interobserver reproducibility of the gravity goniometer and pictures was significant, for the goniometer it was poor. Agreement between the pictures and gravity goniometer assessment of the range-of-motion was good. None of these two methods was significantly correlated with the goniometer assessment. CONCLUSION: For reproducibility of measurement of range-of-motion in the forearm the gravity goniometer is the best method, although the measured values were 5 % greater. PMID- 11898069 TI - [Long-term results of posterior correction and fusion of scoliosis using the Cotrel-Dubousset instrumentation]. AB - AIM: Radiographic analysis of long-term results following CDI. METHODS: Retrospective analysis of radiographs of 43 patients with idiopathic right thoracic scoliosis with an average follow-up of 82 months. Extensive radiographic analysis with special regard to curve correction in the frontal and sagittal planes. RESULTS: The average Cobb-angle of the preoperative primary curve was 61.6 degrees (min.: 40 degrees, max.: 84 degrees ), the correction postoperatively was 51.1 % representing 29.9 degrees. Due to the loss of correction of 7.1 degrees, the overall outcome was 39.9 % by the time of follow up. More than 90 % of the loss of correction occurred within the first 2 years. The lumbar secondary curve correction was 47.2 % (preop.: 37.5 degrees; postop.: 19.8 degrees ). Average fusion length was 11 segments, fusion usually ended two vertebrae below the end vertebra. Translation of the apex was corrected by 38.8 % (preop.: 4.9 cm; postop.: 3.0 cm), tilt of the last instrumented vertebra by 44.2 % (preop.: 18.2 degrees; postop.: 10.16 degrees ). The amount of derotation was negligible (preop.: 24.1 degrees; postop.: 22.6 degrees ). The readjustment of a preoperative pathologic sagittal profile, meaning a thoracic hypokyphosis, was successful in 12 out of 15 cases. Blood loss, duration of operation and complications were documented. CONCLUSION: CD-Instrumentation in scoliosis surgery offers a long-lasting suffcient correction of the fronal and a good correction of the sagittal plane. Stability of correction is achieved 2 years after operation. PMID- 11898070 TI - [Cytokine profile of a human bone marrow cell culture on exposure to titanium aluminium-vanadium particles]. AB - INTRODUCTION: The purpose of this study was to prove the effect of wear particles, especially Tivanium, in the mechanism of the aseptic loosening of total joint prostheses. MATERIALS AND METHODS: Therefore, human bone marrow cell cultures were incubated with titanium-aluminium-vanadium particles of different concentrations which were added after the seventh day of culture (10(9), 10(8), 10(7), 10(6) particles per ml medium). From this time starts the real culture period (2 weeks). During these two weeks the medium was changed and the supernatants were sampled. Using an ELISA the cytokine levels of interleukin-6, interleukin-1beta, TNF-alpha and LDH were measured approximately every second day (1, 3, 6, 8, 10, 14). As a marker for toxicity the activity of LDH was determined. RESULTS: Incubation of a human bone marrow cell culture with titanium aluminium-vanadium particles led to a maximum release of interleukin-6, interleukin-1beta, and TNF-alpha at high particle concentration (10(9) particles per ml medium). An increase of interleukin-1beta was only detectable at particle concentrations of 10(9) per ml medium. Exposure of the human bone marrow cell culture to titanium-aluminium-vanadium particles was toxic for high particle concentrations (10(9) particles per ml medium), as reflected by release of the intracellular enzyme LDH. DISCUSSION: This study shows the ability of tivanium wear particles in a human bone marrow cell culture to induce a signfically higher release of proinflammatory and osteolytic mediators which are responsible for the aseptic loosening of prosthesis and the problem of revisions. In comparison to other cell studies, our results were explained by the human bone marrow cell culture. The human bone marrow is the real effector tissue source "in situ" because the prosthesis is localised intramedullarly. PMID- 11898071 TI - [Histopathological investigations in soft tissue surrounding metallic orthopaedic implants by extravasated eosinophil leukocytes]. AB - INTRODUCTION: The amount of extravasated eosinophil leukocytes in soft tissue surrounding a metallic orthopaedic implant indirectly allows conclusions about the allergenicity of the implant. As yet the implications of the distance of the biopsy from the implant have not been studied. METHODS: In a prospective study we histocytologically evaluated soft tissue biopsies gained during routine removal of orthopaedic implant material in 81 patients. By means of a quantitative and semiquantitative method, material directly adjacent to and 1 cm distant from the implant was examined for eosinophil leukocytes. RESULTS: In respect to the number of eosinophil granulocytes, the two groups showed no significant discrepancies (p = 0.55). lnterestingly, correlation analysis revealed a significant difference in the number of eosinophiles between implant-adjacent and -distant tissue in some individuals. Consecutive semiquantitative analysis could show that there was no correlation concerning the number of eosinophiles between results gained at 1 cm distance from the implant and material in the direct vicinity of the biomaterial. CONCLUSION: Our results suggest that when utilising a histocytological method to determine metal sensitivity of a biomaterial on surrounding soft tissue, not only material directly adjacent to the implant but also at a distance of approximately 1 cm should be evaluated. PMID- 11898072 TI - [Semiquantitative analysis of bone regeneration on porous metallic wire-mesh specimens with or without HA-coating in comparison to pure titanium in animal experiments]. AB - INTRODUCTION: The aim of this study was to analyse the bone regeneration on porous metallic wire-mesh specimens of pure titanium, uncoated CoCrMb and HA coated CoCrMb in a comparative animal experiment. MATERIAL AND METHODS: Uncoated and hydroxyapatite-coated CoCrMb (one-sided) specimens were tested in an animal experiment. The statistical interpretation was done macroscopically as well as microscopically. RESULTS: With regard to the histological and quantitative evaluation, it has been shown that the proximal coated CrCoMb implants were superior to the other specimen. The bone regeneration with the most distinctive extent of osseointegration has been seen by the HA-coated CrCoMb specimen. The uncoated distal CoCrMb specimen demonstrated the worst results. DISCUSSION: This study shows that specimens of titanium and uncoated CrCoMb implanted at the proximal metaphysis deliver comparable effects on the bone regeneration and osseointegration. The best results with regard to the bone regeneration and osseointegration were seen with the implanted hydroxyadaptite-coated CrCoMb specimen. In addition, it has been observed that the bone regeneration and osseointegration of the specimen depends on the place of implantation. PMID- 11898074 TI - [Strengthening of lumbar extensors (MedX)--therapy in chronic backache--an overview and meta-analysis]. PMID- 11898073 TI - [Induction of arthritis in healthy knee joints after intra-articular injection of the proteolytic enzyme elastase - An experimental investigation in the rabbit]. AB - AIM: Humoral aspects are being discussed in the initiation of arthritis. Therefore, the effects of the proteolytic enzyme elastase on the cartilage of knee joints in rabbits have been investigated. The enzyme was evaluated using activities comparable to those in post-traumatic knee joint hemarthrosis in humans. METHOD: Polymorphonuclear leukocyte elastase was injected into one of the knee joints of 10 rabbits. In 5 animals (first study group), joints were then immobilized with a cast for 6 weeks. In the other 5 (second study group), no immobilization was applied. In the first zero group (2 animals), 0.9 % NaCl was injected intra-articularly without immobilization, whereas in the second zero group (2 anmals) knees were immobilized for 6 weeks without prior injection. Thus, the effect of immobilization could be evaluated additionally. Joint specimens were then examined histologically and electron microscopically. RESULTS: There was clear evidence of elastase having severe destructive effects on cartilage regardless of additional joint-immobilization. In neither zero group was there prearthritic damage to the cartilage. CONCLUSION: To prevent the initiation of cartilage damage by humoral factors, early elimination of the pathological intra-articular effusion is necessary. PMID- 11898075 TI - [Economical analyses in interventional radiology]. AB - Considerations about the relation between benefit and expenses are also gaining increasing importance in interventional radiology. This review aims at providing a survey about the published data concerning economical analyses of some of the more frequently employed interventions in radiology excluding neuroradiological and coronary interventions. Because of the relative scarcity of literature in this field, all identified articles (n = 46) were included without selection for methodological quality. For a number of radiological interventions the cost effectiveness has already been demonstrated, e. g., PTA of femoropopliteal and iliac artery stenoses, stenting of renal artery stenoses, placement of vena-cava filters, as well as metal stents in malignant biliary and esophageal obstructions. Conflicting data exist for the treatment of abdominal aortic aneurysms. So far, no analysis could be found that directly compares bypass surgery versus PTA + stent in iliac arteries. PMID- 11898076 TI - [3D real time contrast enhanced ultrasonography,a new technique]. AB - While 3D sonography has become established in gynecology, abdominal applications have been mainly restricted to case reports. However, recent advances in computer technology have supported the development of new systems with motion detection methods and image registration algorithms - making it possible to acquire 3D data without position sensors, before and after administration of contrast enhancing agents. Hepatic (and also splenic) applications involve the topographic localization of masses in relation to the vessels, e.g. hepatic veins and portal vein branches prior to surgical procedures (segment localization). 3D imaging in the characterization of liver tumors after administration of contrast enhancing agents could become of special importance. We report on the first use of 3D imaging of the liver and spleen under real time conditions in 10 patients, using contrast enhanced phase inversion imaging with low mechanical index, which may improve the detection rate and characterization of liver and splenic tumors. PMID- 11898077 TI - [Dynamic MRI of the lumbar spine for the evaluation of microcirculation during anti-angiogenetic therapy in patients with myelodysplastic syndromes]. AB - PURPOSE: Evaluation of MRI perfusion parameters of the lumbar spine in patients with myelodysplastic syndromes (MDS) to determine the vascularisation and anti angiogenetic effects of thalidomide therapy. MATERIAL AND METHODS: In 20 healthy normal persons and 28 MDS patients a dynamic contrast-enhanced MRI (d-MRI) of the lumbar spine was performed. After the initial d-MRI-investigation 24 of the 28 MDS patients received an anti-angiogenetic therapy with thalidomide. With an average of 4.2 months after the beginning of therapy a d-MRI-follow-up examination in 9 of these patients was performed. The amplitude and exchange-rate constant were calculated and a statistical comparison of these values between healthy persons and MDS patients as well as a correlation with the clinical course was executed. RESULTS: Compared with the normal controls the MDS patients showed a higher amplitude (normal persons: 14.4 +/- 5.2, MDS: 24.8 +/- 8.1) and exchange-rate constant (normal persons: 0.124 +/- 0.042, MDS: 0.136 +/- 0.036). In 7 of 9 MDS patients undergoing thalidomide therapy a reduction of the amplitude and exchange rate constant values was evident in the d-MRI follow-up examinations. Clinically these patients showed a therapy response with complete or partial disease remission. CONCLUSIONS: In MDS patients significantly higher d MRI parameters can be demonstrated than in normal persons. Under anti angiogenetic treatment these values decrease in case of a response to therapy. Thus, d-MRI seems suitable for the evaluation of anti-angiogenetic therapy effects. PMID- 11898078 TI - [Studies on image quality, high contrast resolution and dose for the axial skeleton and limbs with a new, dedicated CT system (ISO-C-3 D)]. AB - PURPOSE: Evaluation of 3D-CT imaging of the axial skeleton and different joints of the lower and upper extremities with a new dedicated CT system (ISO-C-3D) based on a mobile isocentric C-arm image amplifier. MATERIAL AND METHODS: 27 cadaveric specimes of different joints of the lower and upper extremities and of the spinal column were examined with 3D-CT imaging (ISO-C-3D). All images were evaluated by 3 radiologists for image quality using a semiquantitative score (score value 1: poor quality; score value 4: excellent quality). In addition, dose measurements and measurements of high contrast resolution were performed in comparison to conventional and low-dose spiral CT using a high contrast phantom (Catphan, Phantom Laboratories). RESULTS: Adequate image quality (mean score values 3 - 4) could be achieved with an applied dose comparable to low-dose CT in smaller joints such as wrist, elbow, ankle and knee. A remarkably inferior image quality resulted in imaging of the hip, lumbar and thoracic spine (mean score values 2 - 3) in spite of almost doubling the dose (dose increased by 85 percent). The image quality of shoulder examinations was insufficient (mean score value 1). Phantom studies showed a high-contrast resolution comparable to helical CT in the xy-axis (9 lp/cm). CONCLUSION: Preliminary results show, that image quality of C-arm-based CT-imaging (ISO-C-3D) seems to be adequate in smaller joints. ISO-C-3D images of the hip and axial skeleton show a decreased image quality, which does not seem to be sufficient for diagnosing subtle fractures. PMID- 11898079 TI - [Imaging diagnostics of the wrist: MRI and Arthrography/Arthro-CT]. AB - PURPOSE: To evaluate the use of magnetic resonance imaging (MRI) compared with arthrography and arthro-CT (AG/ACT) in patients with wrist pain. METHODS: MRI and arthrography/arthro-CT (AG/ACT) of the wrist joint were retrospectively evaluated in 346 patients over a three-year period. Imaging findings were correlated to surgical results (n = 78) or clinical course in an at least 6-month follow-up. RESULTS: For tears of the triangular fibrocartilage, arthrography, arthro-CT, and MRI demonstrated a sensitivity and specificity of more than 0.96. Only the positive predictive value was superior for arthrography/arthro-CT (0.99 and 0.98, respectively) compared with MRI (0.94). Arthrography was superior for functional diagnosis of scapho-lunate ligament tears (n = 25). Ulno-lunate and ulno triquetral ligament defects were demonstrated more exactly by arthrography. Traumatic osseous defects, particularly scaphoid fractures (n = 33) and avascular necrosis (n = 17), were better diagnosed using MRI. CONCLUSION: For suspected lesions of the triangular fibrocartilage complex, AG/ACT is slightly more reliable than MRI. However, MRI was found to be highly accurate in diagnosing TFC tears, and is superior to AG/ACT in detecting traumatic and vascular lesions of the wrist. PMID- 11898080 TI - [Preliminary results of coronary artery examination using a 3D-navigator sequence on a high performance MR system]. AB - PURPOSE: To evaluate MR coronary angiography using a three-dimensional navigator echo sequence on a MR scanner with a high performance gradient system. MATERIAL AND METHODS: Five healthy volunteers were examined with a 1.5 Tesla MR system, using a high performance prototype gradient system (peak amplitude 50 mT/m, rising time 600 mus). For imaging, a navigated gradient-echo pulse sequence with an in-plane resolution between 0.63 x 0.63 and 0.78 x 0.78 mm2 was used. Per patient two overlapping slabs were acquired. The number of visualized coronary artery segments was estimated (AHA classification). In addition, signal-to-noise measurements were performed in the ascending aorta at the level of the proximal right and left coronary arteries. RESULTS: In all volunteers the left main, the right coronary artery up to segment 3, the LAD up to segment 8 and the RCX with segment 11 and 13 were clearly visualized. The average signal-to-noise value at the level of the right coronary artery was 11.4 +/- 5.0, at the level of the left coronary artery 12.3 +/- 4.5. One volunteer was measured with an in-place resolution of 0.63 x 0.63 mm2. This resulted in a too low signal-to-noise ratio so that an adequate assessment of coronary arteries was no longer possible. CONCLUSION: 3D-MR coronary angiography using the navigator technique is limited by the signal-to-noise ratio. PMID- 11898081 TI - [Evaluation of myocardial perfusion reserve in patients with CAD using contrast enhanced MRI: a comparison between semiquantitative and quantitative methods]. AB - OBJECTIVE: Comparison between two semiquantitative methods and a quantitative evaluation of myocardial blood flow (MBF) for assessment of myocardial perfusion reserve (MPR) in patients with CAD. MATERIAL AND METHODS: 9 patients with coronary stenoses > 50 % were examined with an ECG-gated Saturation Recovery Turbo FLASH sequence by using Gd-DTPA as contrast agent (CA). The entive measurements were performed both during rest and hyperemia induced by adenosine. The up-slopes of the signal-time S(t) curves in the myocardium and left ventricular (LV) cavity were evaluated by a linear fit. MPR was calculated from the original up-slopes of the myocardial S(t) curves and from the up-slopes, which were normalized to the up-slopes of the LV S(t) curves, respectively. For quantification of MBF values, the mathematical model MMID 4 was used and MPR was evaluated from the MBF values. RESULTS: With all tested methods, MPR was reduced in myocardial regions subtended by arteries with stenoses >/= 70 % compared with remote regions. With MMID 4 and the normalized up-slope method, differences between severe ischemic and remote regions were statistically significant. CONCLUSION: The up-slope method with normalization and quantification with MMID 4 are more sensitive methods to differentiate between remote and ischemic myocardium than the up-slope method without normalization. PMID- 11898082 TI - [Comparison between fast MR flow quantification in breathhold technique in ascending aorta and pulmonary trunc with right and left ventricular cine-MRI for the assessment of stroke volumes in healthy volunteers]. AB - PURPOSE: Evaluation of accordance and reproducibility of the stroke volume assessment of fast flow measurement in breathhold technique in the ascending aorta and pulmonary trunc with fast right (RV) and left ventricular (LV) cine MRI. MATERIAL AND METHODS: The correlation an accordance of stroke volumes (SV) were evaluated by flow measurements in the ascending aorta and pulmonary trunc and RV and LV volumetric cine-MRI in 33 cardiac healthy volunteers. Interobserver and intraobserver variability of the different measurement methods and locations were checked in 10 cardiac healthy volunteers. RESULTS: With respect to the early systolic flow and the extrapolation of the endiastolic flow rate the difference in SV was smaller than 1,5 ml/m(2) for all possible combinations. The interobserver and intraobserver variabilities of flow measurements were significantly lower by approximately a factor of 2.5 than those of RV and LV cine MRI. CONCLUSION: The accordance of fast prospective triggered flow measurements in the ascending aorta and pulmonary trunc with RV and LV cine-MRI can be achieved with reasonable limits of agreement. Flow measurements are more reproducible than cine-MRI. PMID- 11898083 TI - [Simple localization of peripheral pulmonary nodules - CT-guided percutaneous hook-wire localization]. AB - BACKGROUND: Video-assisted thoracoscopic surgery (VATS) is an alternative approach to small intrapulmonary nodules, if transbronchial or percutaneous biopsy have failed. We investigated the feasibility and effectiveness of the percutaneous CT-guided placement of hook-wires to localize such nodules before video-assisted thoracoscopy. SUBJECTS AND METHODS: 19 patients with new by diagnosed intrapulmonary nodules underwent CT-guided hook-wire localization by application of a X-Reidy -Set (Cook, Inc., Bjaeverskov, Denmark). The average age of the patient was 63 years (range: 19 - 80 years), the mean distance between the nodule and the pleura visceral was 7.58 mm (range: 0 - 25 mm) and the mean diameter was 11.58 mm (range: 5 - 25 mm). After localization, the patients underwent a VATS resection of the lesion within a mean time of 30 min (range 10 - 48 min). RESULTS: In all cases, resection of the nodules was successful. In 4 older patients the marking was complicated by poor cooperability. At the end of manipulation the end of the hook was distanced from the nodule. But also in these cases, resection was successfully performed. 8 patients developed an asymptomatic pneumothorax: 5 of them in a minor (max. 1.5 cm rim), three of them in a moderate (max. 3 cm rim) dimension. In 4 patients, in whom the tumor was hit directly by the needle, local bleeding occurred. In one case, haemoptoe was present. In no patient did a dislocation of the hookwire-system occur. CONCLUSION: CT-guided placement of a hook-wire system is a simple and reasonable procedure which facilitates safe VATS resection of small pulmonary nodules. PMID- 11898084 TI - [Sedation analgesia in interventional radiology]. AB - PURPOSE: Development of a save and effective protocol for analgosedation of patients undergoing painful interventional procedures. MATERIAL AND METHODS: In a prospective trial a consecutive series of 72 adult patients underwent analgosedation during painful interventions. A radiologist performed the analgosedation, the patients received a combination of a shortly effective piperidine derivative (Alfentanil [Rapifen(R)]; 7.5 - 15 microgram/kg body weight) and Benzodiazepine (midazolam [Dormicum(R)]; 20 microgram/kg body weight). After pre-procedure oxygenation patients were continuously monitored. Pain and discomfort were scored using an established visual-analog pain score (0 10). A control group (n = 24) had received midazolam, pentazocine or fentanyl according to the study protocol. RESULTS: All procedures could be carried out by an interventional radiologist and a nurse and/or technologist only. In 69/72 cases adequate analgosedation could be achieved. Injection of alfentanil was titrated, with a rapid onset and short acting effect of the analgesia. Patients reported an average pain score of 2.6 vs. 4.5 in the control group. Over 55 % experienced no or mild pain (score 0 - 3), in the control group only 8 % reached this level. CONCLUSION: A combination of shortly effective alfentanil and midazolam allows interventional radiologists to perform major procedures alone under effective analgosedation. This medication scheme is superior to the medication upon demand. PMID- 11898085 TI - [Percutaneous radiofrequency ablation of liver malignancies: first experience with a 200-W radiofrequency generator]. AB - PURPOSE: To evaluate the effectiveness of a percutaneous radiofrequency ablation device, consisting of a LeVeen probe and a newly developed 200-W radiofrequency generator. MATERIAL AND METHOD: In 20 patients (9 male, 11 female), 30 primary and secondary liver malignancies were treated by CT-guided percutaneous radiofrequency ablation. The patients were separated in a potentially curable (n = 10) and a palliative (n,= 10) treatment group. The treatment was performed using an umbrella-shaped LeVeen probe with a diameter of 3.5 cm or 4 cm and a 200 W generator (Radiotherapeutics, Mountain View, CA, USA). All interventions were performed under general anesthesia by a standardized treatment protocol. The size of the lesions treated ranged from 2.2 x 2 x 2 cm to 10.8 x 10.1 x 10.5 cm. RESULTS: In all patients, the correct placement of the probe and radiofrequency ablation was technically successful. In two patients more than one treatment session was necessary to achieve the intended treatment result. In all patients of the potentially curable group a complete tumor necrosis could be achieved, whereas in the palliative treatment group complete tumor necrosis was only achievable in one patient. The achieved necrosis size ranged from 2.4 x 2.8 x 2.5 cm to 9.2 x 8 x 10.5 cm. In one patient a colon fistula was observed 4 weeks after treatment. No further complications were observed. The follow-up period ranges from 1 to 15 months (mean 4.5 months). CONCLUSION: By use of a powerful generator in combination with a LeVeen probe percutaneous treatment of even large liver tumors is possible. Radiofrequency ablation is a safe procedure for the minimal-invasive therapy of non resectable liver neoplasms. PMID- 11898086 TI - [The value of magnetic resonance imaging (MRI) for the follow-up of patients with transjugular intrahepatic portosystemic shunts (TIPS)]. AB - PURPOSE: To prospectively determine the value of magnetic resonance imaging (MRI) with flow quantification in the portal vein for the follow-up of patients with transjugular intrahepatic portosystemic shunt (TIPS). METHODS: Thirty-six patients with TIPS (23 m, 13 f) were evaluated with MR of the liver parenchyma and quantification of flow in the portal vein. MR examinations were correlated with Doppler sonography and conventional angiography including measurement of the portal pressure gradient (PPG). In cases of re-interventions (dilatation/stent application) additional examinations with MRI and Doppler sonography were performed. RESULTS: MR flow measurements in the portal vein correlated with Doppler sonography (r = 0.69) whereas no correlation of both methods with the PPG was found. No threshold velocity in the portal vein could be determined to predict shunt stenosis. All shunt occlusions (n = 5) were diagnosed correctly by MRA. Thirty measurements before and after successful angiographic interventions revealed a significant increase in portal flow velocity and a significant decrease of the PPG. Magnetic resonance images enabled a reliable detection of procedural complications (parenchymal bleedings, n = 31; extra and subcaspular hematomas, n = 2 each) and newly occurring hepatocellular carcinomas (n = 2) in the follow-up period. CONCLUSION: Magnetic resonance imaging in the follow-up of TIPS enables a morphological assessment of the liver and an accurate velocity mapping, but is not suited to predict shunt dysfunction as a single method. PMID- 11898087 TI - [First experiences with a 6F-sheath compatible self-expanding nitinol stent]. AB - BACKGROUND: To evaluate the efficacy, safety and limitation of a new 6F compatible nitinol stent (Dynalink(TM). MATERIAL AND METHODS: We treated 50 patients (17 women, 33 men, mean age 72 +/- 8 years) by implanting 80 Dynalink(TM)-stents into 75 vessels during a 6-month period. Target lesions were: iliac artery: 25, femoral artery: 31, popliteal artery: 16, femoro-popliteal bypass: 5, subclavian vein: 3. Preinterventional Rutherford classifications: Class 1 : 3 legs (5 %), class 2 : 27 legs (51 %), class 3 : 16 legs (31 %), class 4 : 2 legs (4 %), class 5 : 5 legs (9 %). 40 % stents each were implanted ipsilateral, 60 % cross-over. RESULTS: All interventions were successful regardless of a sometimes anatomically difficult access to the lesion. The device was characterised by a high flexibility and radial force and the stent did not shorten. COMPLICATIONS: One distal stent dislocation during placement occurred, no puncture site complication. The mean diameter stenosis was reduced from 91 +/- 10 % (75 - 100 %) to 4 +/- 8 % (0 - 30 %). The ankle-brachial index was improved from 0.46 +/- 0.22 to 0.75 +/- 0.23 (p < 0.001). Post-interventional Rutherford classifications: Class 0 : 43 legs (81 %), class 1 : 5 legs (4,5 %), class 5 : 5 legs (4,5 %). CONCLUSIONS: The new 6F-sheath compatible nitinol stent is characterised by a good flexibility, radial force, and a lack of shortening. By the reduction of the diameter of the device to 6F, the potential risk of a local bleeding complication may be reduced and 6F sealing devices will be usable. Disadvantages are the 0.018 inch guide-wire lumen and the limited stent sizes. PMID- 11898088 TI - [Atypical mycobacteriosis of the breast: MR diagnosis]. PMID- 11898089 TI - [Traumatic occlusion of the iliac artery after a merry-go-round ride]. PMID- 11898090 TI - [CT diagnosis of asymptomatic cor triatriatum sinistrum]. PMID- 11898091 TI - [Concerning the article by N. Holzknecht et al. (Fortschr Rontgenstr 2001; 173:612). Evaluation of an electromagnetic virtual guide system (CT-guide) for performing CT interventions]. PMID- 11898095 TI - "On the Way to Higher Skills" PMID- 11898096 TI - Mortality in Obstructive Sleep Apnea: Association with Impaired Wakefulness. AB - Sleep fragmentation from obstructive sleep apnea (OSA) is correlated with a shortened sleep latency on the Maintenance of Wakefulness Test (MWT) and the Multiple Sleep Latency Test. Whether impairment of wakefulness is associated with increased mortality in OSA patients is unknown. We evaluated survival over an average timespan of 7.5 years from the date of diagnosis in a consecutive series of 322 OSA patients who had undergone nocturnal polysomnograpy and the MWT. Evaluable survival data were obtained in 142 patients. Twenty two had died. Deaths were predominantly due to cardiovascular disease. A comparison of the demographic and sleep study data between the alive and dead groups was significant for differences in MWT sleep latency and in age at time of diagnosis. The MWT mean sleep latency, when adjusted for age, was significantly shortened in the dead patients (28 +/- 11 min vs. 21 +/- 10 min, p < 0.005). Also, there was a significant decrease in survival in the patients whose MWT mean sleep latency was less than 20 min. These findings demonstrate an association between impairment of wakefulness and long-term mortality in OSA patients. This association was not evident for the other measures used to assess OSA severity. PMID- 11898097 TI - Persistence of Apnea in Wakefulness in a Patient with Postradiation Pharyngitis. AB - We report on a patient with the onset of recurrent nocturnal awakenings associated with postawakening stridor with onset a few weeks after receiving radiation therapy to the neck. The onset of nocturnal stridor was also accompanied by complaints of snoring and excessive daytime sleepiness. Stridor did not occur during daytime wakefulness. Nocturnal polysomnography (NPSG) recorded with a calibrated pneumotachometer demonstrated snoring and severe obstructive sleep apnea (OSA) with a apnea/hypopnea index of 51 events/hr. One apneic episode persisted for 17 sec after the onset of wakefulness as evidenced by standard NPSG scoring criteria for arousals. With this event, video monitoring revealed the patient abruptly sitting upright and clutching his throat and auditory recording demonstrated stridorous sounds. During wakefulness endoscopy revealed moderate edema and erythema of the supraglottic region, epiglottis, palatine tonsils, and false and true vocal cords. Vocal cord function appeared normal. This case report represents the observation of two rare findings in a single patient, persistence of apnea in wakefulness, and OSA onset following neck irradiation. We review the literature on the persistence of apnea in wakefulness and discuss possible mechanisms for its occurrence in this patient. PMID- 11898098 TI - Executive Functions in Persons with Sleep Apnea. AB - Seventeen patients with sleep apnea syndrome [SAS, Respiratory Disturbance Index (RDI) = 12-85] were compared with 16 normal controls (RDI < 7) on neuropsychological tests of executive functions, a domain in which SAS patients have been suggested to have deficits. SAS patients demonstrated greater deficits in the retrieval of information from semantic memory (Controlled Oral Word Association task) and in shifting responses in the face of error (Wisconsin Card Sort Test), but differences in working memory were not observed. Eliciting deficits in cognitive executive functions in SAS may require more sensitive measures than are typically used in neuropsychiatric research. PMID- 11898099 TI - The Origin of Pharyngeal Obstruction during Sleep. AB - This paper traces the development of the adult human pharynx through evolution, comparative anatomy, and development from infant to adult. Based on phylogeny and otogeny, an hypothesis for the occurrence of obstructive sleep apnea in the mature human is presented. PMID- 11898101 TI - Severity of Illness and Treatment Thresholds: Managed-Care Strategies in Sleep Medicine. AB - The objectives of this article are: (1) to distinguish functional from physiological criteria for illness; (2) to articulate threshold approach to our diagnostic strategies; and (3) to list the functions of sleep medicine that give valued patient care systems. PMID- 11898100 TI - Disorders of Sleep and Breathing during Sleep in Neuromuscular Disease. PMID- 11898102 TI - Fat and Prejudice. PMID- 11898103 TI - Who is Mr. Z? PMID- 11898104 TI - Over the Counter, Under the Blanket: Snoring Treatment from the Supermarket. PMID- 11898105 TI - Sleep Apnea in Moderate-Severe Obese Patients. AB - Obesity induces multiple physiologic changes at the respiratory and circulatory systems level. A study was developed to identify symptoms and signs able to discriminate subjects at high risk of obstructive sleep apnea (OSA) and to evaluate the presence of OSA in a population of obese patients referred to the Clinical Nutrition Service of the Luigi Sacco Hospital for weight loss therapy. Twenty-seven obese patients (14 males, 13 females) without neurologic, cardiac, and lung diseases were measured for height, weight, neck, waist, and hip circumference; a sample of venous blood was taken for hematological data; and were given a pulmonary function test, hemogasanalysis, and full-night polysomnography. Statistical analysis were performed using paired and unpaired StudentOs t test, PearsonOs chi square, and Spearmann Rank correlation; the significance level was set at p<0.05. The results showed hemotological values in the normal range and pulmonary function findings were not different from predicted, but expiratory reserve volume (ERV), as expected in obese subjects, was significantly reduced (p<0.001). Waist, hip, and neck circumference, and waist/hip ratio were 114 +/-14, 118 +/-12, 44 +/-4, and 0.96 +/-0.4 cm respectively. An apnea-hypopnea index (AHI) cutoff value of <15 was used to classify the patients as suffering from OSA: 15 patients (12 males, 3 females, age in years 55 +/-12, body mass index (BMI) kg/m(2) 37 +/-6, AHI 30 +/-12) were OSA and 12 patients were non OSA (2 males, 10 females, age in years 49 +/-20, BMI kg/m(2) 35 +/-2, AHI 3 +/-2). PaO2 and pH were lower and PaCO2 higher in OSA (p<0.05, p<0.01, p<0.05, respectively). Red blood cells (RBC), Hb, and neck circumference were increased in OSA (p<0.05). In OSA patients, S3%, S4% of total sleep time, SaO2% mean of nadir were reduced (p<0.001), and DEF increased (p<0.0001). In obese patients, AHI was correlated with neck circumference (r = 0.74, p<0.0001) and waist/hip ratio (r = 0.48. p<0.01). DEF was correlated with RBC, Hb, Htc% (r = 0.82, 0.71, 0.66, p<0.001). SaO2;% mean of nadir was significantly related to RBC, Hb, and Htc% (r = 0.44, 0.40, p<0.05, respectively). Our data showed a prevalence of OSA in 55% of the obese patients. A significant correlation exists between RBC, Hb, Htc%, with desaturation events frequency (DEF) and SaO2% of nadir indicating that transient, episodic desaturation during sleep is linked to a moderate increase of RBC and Hb found in obese patients with OSA, in contrast to obese, nonOSA patients. The most important result of the present study was the determination that classical symptoms and signs of OSA, such as male gender, neck circumference, waist/hip ratio, RBC, and Hb at the upper limit of normal, are simple inexpensive screening tools, and useful predictors of sleep-disordered breathing and discriminate the individuals with higher risk of OSA. PMID- 11898106 TI - A Randomized Double-Blind Placebo-Controlled Evaluation of the Safety and Efficacy of a Natural Over-The-Counter (OTC) Medication in the Management of Snoring. AB - More than 40 million American adults snore. Habitual snoring afflicts 44% of adult males and 28% of females.(1) Uncomplicated snoring is generally due to vibration of the palatal soft tissues or the tongue base, causing intermittent airway obstruction. Loudness is correlated with the degree of vibration and/or obstruction. The tendency, frequency, duration, intensity, and sequelae of snoring are influenced by myriad structural, physiological, environmental and pharmacological factors. Uncomplicated, nonapneic snoring is treated in a wide variety of ways, ranging from self-help methods, such as positional therapy, to laser surgery. The purpose of this report is to evaluate the safety and efficacy of a natural medication for snoring in a randomized double-blind placebo controlled trial. The treatment is significantly more effective than placebo. Neither side effects nor intolerance to the product was reported. PMID- 11898107 TI - UPPP or LAUP: Is This All Surgeons Should Talk About? AB - Snoring is a nuisance at least! The inspiratory vibrations of pharyngeal soft tissue may exceed 85 dB. For thousands of years snoring has led to social and marital disharmony in many cases. The treatment of the symptom snoring has been of interest for the medical profession for the same time. Today, we know that snoring is the most common presenting symptom of obstructive sleep apnea (OSA). The wider recognition of OSA and its impact on the development of systemic and pulmonary hypertension, cardiac arrhythmias, heart failure and daytime sleepiness has occurred in the last decade. The treatment of OSA has become a medical issue apart from its social aspects. PMID- 11898109 TI - ENT Medicine and Otolaryngology Guidelines of the German Society of Otolaryngology-Head and Neck Surgery. PMID- 11898108 TI - Meta-Communication and Sleep Medicine. PMID- 11898110 TI - Diagnosis and Treatment of Sleep and Breathing Disorders Symposium (held December 9-11, 1998, in Grenoble, France): Meeting Highlights. PMID- 11898111 TI - Methodology and Clinical Decisions. PMID- 11898112 TI - Sleep and Breathing in Recreational Climbers at an Altitude of 4200 and 6400 Meters: Observational Study of Sleep and Patterning of Respiration During Sleep in a Group of Recreational Climbers. AB - Background: The increasing popularity of mountain climbing will result in greater numbers of the general population being at risk for the disturbances known to occur with altitude exposure. Methods: Observations of sleep and breathing were made in 6 healthy travellers (5 males and 1 female, 38 to 62 years of age) before, during, and after a recreational climb. We modified a portable seven channel polygraph to record sleep state, oxygen saturation, respiratory movements, body position, and oronasal airflow 4 weeks prior to the expedition at home (500m), at base camp (4200m) and in 3 climbers at 6400m. All had a repeat study at 500m altitude 4 weeks after the expedition. Results: For the group, the total number of obstructive apneas and hypopneas (OA/H) at night increased from 36 at home to 68 at base camp over a one night recording. Separately counted central apneas and hypopneas (CA/CS) increased from 6.7 to 45. In one climber, who had a history of recurrent snoring and observed apneas at home, the number of apneas increased from 201 at 4200m to 322 at 6400m, whereas in 2 climbers measured at 6400m, all apneas decreased. The total sleep time (TST) increased in all 6 climbers by 10% at base camp in comparison to home records. In the 3 climbers attaining an altitude of 6400m, the REM (rapid eye movement) sleep declined by 10% compared to the record at 4200m. Conclusion: Respiratory disturbances at low altitude are amplified by exposure to high and extreme altitude. In those without symptoms of sleep apnea, significant physiologic alterations will occur at high altitude but at extreme altitude regular ventilation is re-established. PMID- 11898113 TI - Snoring and Sleep Apnea in Obese Adolescents: Effect of Long-term Weight Loss Rehabilitation. AB - Objective: To test the effect of a long-term weight loss rehabilitation program in extremely obese adolescents on breathing parameters during sleep. Methods: Thirty-eight extremely obese [mean body mass index (BMI) 45.3 +/- 7.9kg/m(2)] adolescents participated during a three- to nine-month period in an inpatient weight loss rehabilitation in a specialized long-term rehabilitation center. Breathing parameters were registered via a seven-channel portable screening device. Body weight and arterial blood pressure were measured before and after the long-term treatment. Results: Mean BMI decreased from 45.3 to 35.8 (p < 0.001), mean diastolic blood pressure decreased from 89 mmHg to 81 mmHg (p = 0,002). Nine patients had a RDI of >/=5 and 30 patients a RDI of <5; the mean RDI decreased from 4.08 to 3.27 (n.s.). Within the group, the RDI was >/=5 and the mean RDI decreased from 10.3/h to 5.2/h (p = 0.02). The mean SaO2 increased from 93.65 to 95.35% (p = 0.003), lowest SaO2 increased from 72.14 to 73.19% (n.s.) and snoring frequency decreased from 37.56% of total sleep time (TST) to 32.86% of TST (n.s.). Conclusion: A long-term inpatient weight loss program has a positive effect on breathing parameters during sleep in extremely obese adolescents. However, the effect on apneic events and snoring is relatively minor compared to the effect on arterial oxygen saturation. The role of obesity in the origin of respiratory events and snoring in adolescents might be overestimated. PMID- 11898114 TI - Sleep and Breathing at High Altitude. AB - Sleep at high altitude is characterized by poor subjective quality, increased awakenings, frequent brief arousals, marked nocturnal hypoxemia, and periodic breathing. A change in sleep architecture with an increase in light sleep and decreasing slow-wave and REM sleep have been demonstrated. Periodic breathing with central apnea is almost universally seen amongst sojourners to high altitude, although it is far less common in long-standing high altitude dwellers. Hypobaric hypoxia in concert with periodic breathing appears to be the principal cause of sleep disruption at altitude. Increased sleep fragmentation accounts for the poor sleep quality and may account for some of the worsened daytime performance at high altitude. Hypoxic sleep disruption contributes to the symptoms of acute mountain sickness. Hypoxemia at high altitude is most severe during sleep. Acetazolamide improves sleep, AMS symptoms, and hypoxemia at high altitude. Low doses of a short acting benzodiazepine (temazepam) may also be useful in improving sleep in high altitude. PMID- 11898116 TI - The Emperor (and My Patient) Has No Nose. PMID- 11898117 TI - A Model for Continuous Quality Control of Sleep Laboratories. PMID- 11898115 TI - Recent Developments in Oral Appliance Therapy of Sleep Disordered Breathing. AB - Oral appliances are increasingly gaining a place in the treatment of sleep disordered breathing caused by upper airway obstruction. This review of publications since 1995 documents substantial progress in the scientific basis for this therapy. Imaging by several techniques has shown that mandibular advancing oral appliances open the airway in awake and anaesthetized subjects, creating the presumption that this effect is maintained in sleep. Three controlled cross-over treatment trials have shown that patients consistently prefer oral appliance over continuous positive airway pressure therapy, especially when the treatment effect is strong. Appliance design and use indicates a preference for adjustable mandibular advancing appliances. Complications of therapy appear to be infrequent, but evidence for safety of long term use is still limited. Oral appliance therapy can be an effective therapy for sleep disorders caused by upper airway obstruction. Considering the accumulated evidence, it is no longer tenable to label oral appliance therapy an OexperimentalO procedure. PMID- 11898118 TI - The Swivel Syndrome: Negative and Positive Aspects of a Spreading Phenomenon. PMID- 11898119 TI - Delayed Diagnosis of Obstructive Sleep Apnea: Don't Ask, Don't Tell. AB - To define the patterns of referral of adult patients with obstructive sleep apnea syndrome (OSAS) for diagnostic sleep testing, we studied 97 consecutive patients with OSAS (Apnea/Hypopnea Index >/= 20) referred to the Center for Sleep and Ventilatory Disorders, an American Sleep Disorders Association-accredited university sleep center. Chart review and semi-structured patient telephone interview quantified the time between the onset of any major feature of OSAS and referral to the sleep center, as well as the time between the first OSAS complaint to a health care provider and referral to the center. The average time elapsed between first recognition by the patient of a major feature of OSAS to sleep center referral was 87.5 months (range, 1 to 480 months). Only 4% of referrals were made as a result of the clinician eliciting a history of sleep related complaints. Once OSAS-related features were apparent to the clinician, the average time to referral for diagnostic testing was 7.9 months (range, 0 to 128 months). These data suggest that both a lack of reporting of symptoms by OSAS patients and a lack of obtaining appropriate sleep history by health care providers contribute to a significant delay in diagnosis of OSA. PMID- 11898120 TI - Obstructive Sleep Apnea and Hypopnea Efficacy and Safety of a Long-Acting beta2 Agonist. AB - The effect of inhaled long-acting beta2-agonists in obstructive sleep apnea syndrome (OSAS) is unknown, although from the pharmacological point of view both therapeutic and adverse effects need to be considered. The purpose of this study was to obtain data on the efficacy and safety of salmeterol in patients with OSAS. In a randomized, double-blind, placebo-controlled, cross-over study, effects of salmeterol on respiration during sleep and sleep quality were investigated in 20 patients with OSAS. Of these, 4 patients were female, 16 male; the average age was 53.0 +/- 7.8 years, with average body mass index 28.0 +/- 3.0 kg small middle dot m(-2) and average apnea hypopnea index 35.6 +/- 17.8 h(-1). Patients with asthma, chronic obstructive pulmonary disease (COPD), and left heart failure were excluded. Placebo or verum (50 &mgr;g salmeterol) was administered at 7 pm by meter dose inhaler and spacer device. All patients underwent full polysomnography during baseline, placebo, and verum night. Statistical analysis was performed by StudentOs t-test (p > 0.05). Between the placebo and verum there were no differences in total sleep time, sleep stages, apnea index (AI), apnea hypopnea index (AHI), and nadir oxygen saturation. There was, however, 1) a significant deterioration of mean oxygen saturation (SaO2m; placebo 93.1 +/- 2.0 vs. verum 92.5 +/- 2.2%; p = 0.01), 2) of percent of time spent with an oxygen saturation (SaO2) C transversion occurs at the -1 position of the 5' splice junction of intron 7. Two female relatives who are heterozygous for the SLC6A8 mutation also exhibit mild mental retardation with behavior and learning problems. Male patients with the mutation have highly elevated creatine in their urine and have decreased creatine uptake in fibroblasts, which reflects the deficiency in creatine transport. The ability to measure elevated creatine in urine makes it possible to diagnose SLC6A8 deficiency in male patients with mental retardation of unknown etiology. PMID- 11898127 TI - Hereditary spastic paraplegia SPG13 is associated with a mutation in the gene encoding the mitochondrial chaperonin Hsp60. AB - SPG13, an autosomal dominant form of pure hereditary spastic paraplegia, was recently mapped to chromosome 2q24-34 in a French family. Here we present genetic data indicating that SPG13 is associated with a mutation, in the gene encoding the human mitochondrial chaperonin Hsp60, that results in the V72I substitution. A complementation assay showed that wild-type HSP60 (also known as "HSPD1"), but not HSP60 (V72I), together with the co-chaperonin HSP10 (also known as "HSPE1"), can support growth of Escherichia coli cells in which the homologous chromosomal groESgroEL chaperonin genes have been deleted. Taken together, our data strongly indicate that the V72I variation is the first disease-causing mutation that has been identified in HSP60. PMID- 11898133 TI - Opening the door to new opportunities. Reducing cancer mortality and incidence. PMID- 11898134 TI - Brachial plexopathy after treatment for breast cancer. PMID- 11898128 TI - PKHD1, the polycystic kidney and hepatic disease 1 gene, encodes a novel large protein containing multiple immunoglobulin-like plexin-transcription-factor domains and parallel beta-helix 1 repeats. AB - Autosomal recessive polycystic kidney disease (ARPKD) is a severe form of polycystic kidney disease that presents primarily in infancy and childhood and that is characterized by enlarged kidneys and congenital hepatic fibrosis. We have identified PKHD1, the gene mutated in ARPKD. PKHD1 extends over > or =469 kb, is primarily expressed in human fetal and adult kidney, and includes a minimum of 86 exons that are variably assembled into a number of alternatively spliced transcripts. The longest continuous open reading frame encodes a 4,074 amino-acid protein, polyductin, that is predicted to have a single transmembrane (TM)-spanning domain near its carboxyl terminus, immunoglobulin-like plexin transcription-factor domains, and parallel beta-helix 1 repeats in its amino terminus. Several transcripts encode truncated products that lack the TM and that may be secreted if translated. The PKHD1-gene products are members of a novel class of proteins that share structural features with hepatocyte growth-factor receptor and plexins and that belong to a superfamily of proteins involved in regulation of cell proliferation and of cellular adhesion and repulsion. PMID- 11898135 TI - Patient age and cancer treatment decisions. Patient and physician views. AB - PURPOSE: The purpose of this study was to examine patient and physician factors influencing the decision to use adjuvant chemotherapy for stage III colon cancer in elderly persons. DESCRIPTION OF STUDY: A cross-sectional mailed population based survey of patients 65 years of age and older who had undergone surgical resection of stage III colon cancer in Colorado between August 1995 and December 1997 were identified by the statewide cancer registry (n = 276) and their treating physicians (n = 232). A questionnaire about the determinants of colon cancer treatment decisions was mailed to all patients for whom physician permission was granted (n = 119). A similar questionnaire was sent to treating physicians. RESULTS: Ninety-two physicians (internal medicine 23%; family medicine 12%; surgery 37%; and oncology 24%) and 67 patients (mean age 75.8 years; 55% women) completed surveys. Fifty-four (80.6%) of the patients had received adjuvant chemotherapy. The major determinants of receiving adjuvant chemotherapy were having seen an oncologist (P = .003), being younger (P = .003), and being married (P = .021). After controlling for other potential influences, only having seen an oncologist (odds ratio 8.0; confidence interval 1.5-43.1) remained significantly associated with the receipt of chemotherapy. Physicians were more likely than patients to rank comorbid conditions (39.1% versus 3.0%, respectively; P < .001) and the medical literature (20.7% versus 4.5%, respectively; P = .004) as important factors in making treatment decisions, while patients were more likely than physicians to rank physician opinion (73.1% versus 26.1%, respectively; P = .001), family preference (31.3% versus 9.8%, respectively; P = .001), and family burden (10.4% versus 2.2%, respectively; P = .038). CLINICAL IMPLICATIONS: In this elderly population, patient age is not recognized by patients or physicians as affecting the decision to use adjuvant chemotherapy. Other biologic and social factors are important, however, and the perspectives of physicians and patients differ regarding their relative importance. PMID- 11898136 TI - Caregiver responses and needs. An ambulatory bone marrow transplant model. AB - PURPOSE: This longitudinal, descriptive outcomes study was conducted to compare the emotional responses and needs of the caregivers of patients who undergo bone marrow transplantation (BMT) for hematologic malignancies, as observed in an inpatient/outpatient (IPOP) setting with those in an inpatient setting. DESCRIPTION OF STUDY: A convenience sample of 43 caregivers for patients undergoing either autologous or allogeneic BMT was selected from both the inpatient unit (n = 26) and the IPOP ambulatory setting (n = 17). Emotional responses were measured by the Profile of Mood States. The importance and satisfaction of informational, patient-care, and psychological needs were assessed with the Caregiver Needs Survey, a modified version of the Home Cancer Caregiver Needs Survey. Data were collected at six points across the BMT trajectory, from before the procedure to 12 months after. Descriptive statistics were used to report sample characteristics, emotional responses, and needs. Nonparametric statistics were used to compare the IPOP and inpatient caregiver groups, and to identify relationships between emotional responses and needs within groups. RESULTS: IPOP caregivers were found to have significantly less mood disturbance at points before discharge. Both groups showed a decrease in mood disturbance 6 and 12 months after BMT. IPOP caregivers had higher levels of satisfaction of their informational and psychological needs at day 21. Significant correlations between total mood disturbance and satisfaction of informational, psychological, and patient-care needs were found for inpatient caregivers across treatment time points. CLINICAL IMPLICATIONS: These findings support the IPOP model of care as being less emotionally distressing for and better meeting the needs of family caregivers. Specific implications for practice include the importance of caregiver education in the area of patient-care information and of assessment and intervention to meet caregiver psychological needs. Continued evaluation of the impact of changes in care delivery on family caregivers is essential for the provision of comprehensive cancer care. PMID- 11898137 TI - Training oncology camp volunteers. A developmental and strengths approach. AB - PURPOSE: This article examines the training of oncology camp volunteers using a developmental and strengths perspective as a theoretical orientation. The literature on oncology camping, both empirical and anecdotal, is examined and relevant studies cited. OVERVIEW: The selection of volunteers, the benefits of the oncology camp experience, and a training model are discussed. It is the contention of the author that camp volunteers need an orientation for working with the campers, one that incorporates both a developmental framework and a perspective that highlights capacity and strength. Both orientations are explored, and their applications are highlighted. CLINICAL IMPLICATIONS: The training of volunteers is a critical aspect of the oncology camp. In almost all oncology camps, volunteers form the staff complement. As such, their training and orientation to the campers is central to the summer experience for campers, volunteers, and staff. Administrators and camp directors are urged to incorporate some of the suggestions offered in this report to help ensure a positive summer experience. PMID- 11898138 TI - Family history of breast cancer. Impact on the disease experience. AB - PURPOSE: Family history is the most prominent risk factor, besides advanced age, for the incidence of breast cancer among women. This study investigates differences in the experiences of women in the detection, diagnosis, and treatment of early-stage disease. The purpose of this research is to obtain a more comprehensive understanding of the impact of family history on the overall illness experience. DESCRIPTION OF STUDY: Self-report retrospective data obtained from in-depth interviews with a convenience sample of 179 women who had recently received a diagnosis of nonrecurrent stage 0 to IIIA breast cancer are used to compare the experiences of women with and without a family history of breast cancer (FHOBC). The authors examine differences in screening behavior, method of detection, diagnostic processes, treatment decision making, and therapy receipt, and they report the results of bivariate analyses. RESULTS: The results suggest that women with FHOBC have a different disease experience than those without an affected relative. Women with FHOBC were more likely than their counterparts to comply with screening guidelines, to seek more timely care, to consult with specialists, to be influenced by the experiences of others, to feel comfortable with treatment decisions, and to receive adjuvant therapy. CLINICAL IMPLICATIONS: Healthcare providers should be aware that compliance with mammography and therapy guidelines may vary with FHOBC. Because the better health-related behavior reported by women with affected relatives suggests that they may have higher perceived risk, physicians should be sensitive to potentially elevated levels of anxiety, provide accurate information about relative risk, put patient concerns in the proper perspective, and include family members in treatment discussions. Alternatively, women without an FHOBC appear to have less favorable screening, detection, diagnosis, and treatment decision-making behavior. Because family doctors play an important role in the care of these patients, they may need to provide special education and counseling regarding the importance of adherence to screening guidelines, recognition of relevant symptoms, initiation of timely examinations, consultation with cancer specialists, and compliance with treatment recommendations. PMID- 11898139 TI - Complementary therapies. A survey of NCI cancer patient educators. PMID- 11898140 TI - The Internet. Changing the way cancer survivors receive support. PMID- 11898141 TI - Angiogenesis inhibitors in oncology. The research continues. PMID- 11898142 TI - Therapy for cancer of the base of the tongue: managing the side effects. PMID- 11898143 TI - Behavioral interventions for lung cancer-related breathlessness. AB - PURPOSE: The purpose of this report is to present behavioral interventions to assist persons with lung cancer in the management of feelings of breathlessness and, thus, also to enhance their quality of life. OVERVIEW: Breathlessness is a serious symptom that adversely affects the quality of life of persons with lung cancer. A review of the literature points to the value of exercises in assisting patients to breathe more effectively and to manage related anxiety. However, the professional literature frequently does not describe these basic interventions in enough detail to enable oncology professionals to learn them. Instructional materials, found in the popular wellness and self-help literature, are included in this article to more easily facilitate acquisition of these skills. Interventions described include exercises that enhance the use of the diaphragm when breathing and those that help to alter the breathing rhythm and to exhale more effectively. CLINICAL IMPLICATIONS: All oncology professionals should be aware of the importance of breathlessness as a problem that diminishes the quality of life for patients with lung cancer. Addressing breathlessness through the use of psychosocially oriented behavioral interventions can act as an adjunct to the medical management of this debilitating symptom. PMID- 11898144 TI - Symptom profile of nasopharyngeal cancer patients during radiation therapy. AB - PURPOSE: This prospective, longitudinal study was aimed to describe the prevalence, severity, and pattern of symptoms over the course of radiation therapy in persons with nasopharyngeal carcinoma and to explore symptom severity by treatment modality. DESCRIPTION OF STUDY: Thirty-seven patients completed this study, and 46% received chemotherapy before radiation therapy. A self-reported radiation symptom checklist and an objective mucositis assessment tool were used weekly to document oropharyngeal, skin, nose or ear, or more general side effects, and mucositis. RESULTS: Oropharyngeal problems were the most severe complaints during radiation therapy. All patients experienced dry mouth, taste change, difficulty in swallowing, difficulty in opening their mouths, hoarseness, sore throat, and observable mucositis. Most reported moderate-to-severe dry mouth, difficulty in swallowing, and sore throat from weeks 3 through 7. Skin problems were not prominent until week 4. Patients also lost an average of 3.9 kg during the therapy. Sequential chemotherapy and radiation therapy was associated with more severe oropharyngeal problems than radiation therapy alone, but no significant differences in other problems were found. CLINICAL IMPLICATIONS: Despite recognition of the oropharyngeal side effects associated with irradiation, effective management protocols for such symptoms have not been implemented in the studied institution. The frequency and intensity of the symptoms reported indicate an urgent need for increased vigilance about radiation related side effects and pain management. As well, patient education about expected side effects may help mitigate the anxiety that patients experience when these symptoms occur. PMID- 11898145 TI - Screening for second cancers and osteoporosis in long-term survivors. AB - PURPOSE: The purpose of this preliminary study was to describe the extent to which healthcare providers recommend the screening strategies for early detection described by the American Cancer Society (ACS), for breast, gynecologic, and colorectal cancer, and by the National Osteoporosis Foundation (NOF), for osteoporosis, to women who are long-term survivors of breast, ovarian, or endometrial cancer. DESCRIPTION OF THE STUDY: A four-part survey was developed for this study, with the first three parts based on the ACS guidelines for breast, gynecologic, and colorectal cancer screening and the NOF guidelines for osteoporosis screening. The fourth part related to personal characteristics, setting, knowledge, and perceptions of the nurses surveyed. A random sample of outpatient nurses was obtained from the Oncology Nursing Society. Of 668 nurses, 321 (48%) responded (Oncology Certified Nurse (OCN) 68.1%; Advanced Oncology Certified Nurse (AOCN) 16.6%). RESULTS: The most consistently performed screenings that were reported were mammogram (range 74.2-87.7%), professional breast examination (range 73.9-83.7%), and Pap test and pelvic examination (range 61.8-85.2%). The least frequently performed screenings are flexible sigmoidoscopy/colonoscopy (range 20.2-27.7%), bone mineral density testing (range 16.9-19.0%), and height measurement (range 22.5-28.3%). Less than one third of survivors are offered counseling on strategies to promote bone health. CLINICAL IMPLICATIONS: Knowledge of factors associated with osteoporosis and the use of screening strategies for second malignancies in survivors of breast, ovarian, and endometrial cancers can be used to implement activities such as patient education and clinical practice protocols that will increase the use of current screening recommendations. PMID- 11898147 TI - Spirituality, demographic and disease factors, and adjustment to cancer. AB - PURPOSE: The purpose of this study was to examine the relationship between demographic-disease variables, spirituality, and psychosocial adjustment in a heterogeneous sample of patients with cancer. DESCRIPTION OF STUDY: Participants (N = 83) accrued through the Rhode Island Hospital and the American Cancer Society completed questionnaires, and structural equation modeling was used to examine the relationships among disease and demographic factors, spirituality, and psychosocial adjustment to cancer. RESULTS: Of five models tested, a mediational model received the strongest support (chi-square(35)-66.61; P = .005; comparative fix index = .90; root mean square error of approximation = .09), explaining 64% of the variance in psychosocial adjustment. Being a woman, having a longer illness duration, and having a lower disease stage were related to greater levels of purpose in life and religious beliefs, which, in turn, were associated with higher levels of family and social adjustment and psychological health. CLINICAL IMPLICATIONS: The results indicate that spirituality can influence how patients with cancer adjust to their diagnosis and treatment and, thus, support the need for interventions that target spirituality to promote psychosocial adjustment in this population. PMID- 11898148 TI - Does sphincter preservation for rectal cancer compromise survival? PMID- 11898146 TI - Factors influencing oral mucositis in patients receiving chemotherapy. AB - PURPOSE: Oral mucositis is a painful complication of chemotherapy and can greatly affect patients' morbidity and mortality. Findings from two previous studies suggested a decrease in the prevalence of chemotherapy-induced mucositis in patients with solid tumors. The purposes of this study were to follow a large cohort of outpatients to determine the prevalence of mucositis and to identify whether certain clinical factors were significant in the development of mucositis. DESCRIPTION OF STUDY: In this prospective study, a convenience sample of 199 outpatients was followed for three cycles or until mucositis developed. The clinical factors monitored included the following: pretreatment dental examination/repair; initial standard chemotherapy dosage; prophylactic use of colony-stimulating factors; and use of preventive mouthwashes or other prophylactic measures. RESULTS: Oral mucositis developed in 50 patients (25.1%). Prechemotherapy dental examination/repair and initial standard chemotherapy dosage were equivalent among both groups. Of the 48 patients in whom mucositis developed, 10 (20.8%) received prophylactic colony-stimulating factors. Of 134 patients in whom mucositis did not develop, 46 (34.3%) received prophylactic colony-stimulating factors. This difference was statistically nonsignificant. CLINICAL IMPLICATIONS: Differences in the clinical factors investigated could not explain the lower prevalence of oral mucositis among the current patient cohort. The reason for the diminishing prevalence of this side effect remains unclear, and additional parameters, particularly detailed oral hygiene practices, should be evaluated. In the meantime, oncology clinicians should consider the teaching of patients and urging them to use good oral hygiene practices as necessary and potentially preventive measures against chemotherapy-induced mucositis. PMID- 11898149 TI - Resources on colorectal cancer for patients. PMID- 11898150 TI - Temozolomide. A new option for high-grade astrocytomas. PMID- 11898151 TI - Evaluating cesarean deliveries. Exploring ACOG's recommendations to improve outcomes. PMID- 11898152 TI - Curbing cesarean rates. ACOG releases new comprehensive recommendations. PMID- 11898153 TI - Gene mutation linked to primary pulmonary hypertension. PMID- 11898154 TI - More older women getting mammograms. PMID- 11898155 TI - Are Americans aging well? PMID- 11898156 TI - Teen births continue dip; reach record low. PMID- 11898157 TI - Access to health care. APNs, NPs must work to defeat legislative barriers to care. PMID- 11898158 TI - Silicone breast implants. What do we know about their long-term effects? PMID- 11898159 TI - Reproductive genetic technologies. Exploring ethical and policy implications. PMID- 11898160 TI - Providing culturally competent care. Strategies and approaches for perinatal clients. PMID- 11898161 TI - Birds do it, bees do it, let's talk about it. The nurse's role in sexuality counseling. PMID- 11898162 TI - Birthing lessons. One woman's recovery from stroke. PMID- 11898163 TI - Nurses and malpractice. PMID- 11898164 TI - Grieving abortion loss. PMID- 11898165 TI - Blaming nurses for medical errors. Leaders respond to the Chicago Tribune's series. PMID- 11898166 TI - Norplant recalled. PMID- 11898167 TI - NIH reaffirms antenatal corticosteroids for preterm women. PMID- 11898168 TI - Is it time to push? Examining rest in second-stage labor. PMID- 11898169 TI - Planning and designing women's and children's services. Avoid these top 10 problems for best outcomes. PMID- 11898170 TI - Helping hands. Exploring the cultural implications of social support during pregnancy. PMID- 11898171 TI - Birthing faith. Giving a gift of trust and support. PMID- 11898172 TI - Accountability in the health care system. PMID- 11898173 TI - Thalidomide. A new beginning. PMID- 11898174 TI - Meeting the challenges of comprehensive cancer control. PMID- 11898176 TI - Older women unaware of increased breast cancer risk. PMID- 11898177 TI - The patient's child. The forgotten survivor. PMID- 11898178 TI - Ethnic identification with healthcare providers and treatment adherence. PMID- 11898179 TI - Lymphedema in women treated for breast cancer. AB - PURPOSE: The purpose of this report is to discuss the etiologic factors, prevention, and current treatment options for lymphedema, which may occur after breast cancer. OVERVIEW: Lymphedema can develop in the breast cancer patient as a result of the interruption of lymphatic flow from postsurgical, postradiation, and infectious causes. It can present at various points after breast cancer treatment and may range from mild to a seriously disabling enlargement. Because lymphedema is permanent, the goal of treatment options is the control of edema, and a multidimensional approach to care is often needed. CLINICAL IMPLICATIONS: In the past, there has been limited research in this area. Because lymphedema is essentially an incurable condition, healthcare providers should be aware of techniques such as sentinel-node biopsy that could significantly decrease its incidence. In addition, it is important that each patient's treatment include education and prevention precautions. PMID- 11898180 TI - Impact of development on children's mourning. AB - PURPOSE: A qualitative approach was used to analyze the impact of development on the grief responses of 157 children from 88 families to the death of a parent from cancer. DESCRIPTION OF STUDY: Children from age 3 to 17 years were divided by developmental characteristics, derived from interview data, into five development-derived age categories. RESULTS: The responses of children in different categories clarified the impact of development on their distinct expressions of grief, attributes of the parent they mourned, and the parent's tasks in enhancing the reconstitution of the child and the family. CLINICAL IMPLICATIONS: This first attempt to find a way to segregate children into developmentally more homogenous subgroups led to the clarification of patterns of mourning behaviors that are clinically useful. The increased precision of findings, such as the way development affects the child's mourning and the related parental support they need, may help clinicians develop more specific interventions to help children cope with the death of a parent and guide parents in understanding their children's differing responses. The pivotal role of the surviving parent and the tasks required of that parent are important for healthcare professionals to understand and support. PMID- 11898181 TI - Genetic testing for prostate cancer. Willingness and predictors of interest. AB - PURPOSE: As researchers come closer to identifying the genes responsible for prostate cancer, the possibility of genetic testing for men at risk for prostate cancer becomes more likely. This study examined the following: 1) the degree to which men with (n = 43) or without (n = 83) a family history of prostate cancer would be interested in genetic testing; and 2) the degree to which interest in testing was associated with demographic, family history, and psychosocial factors. DESCRIPTION OF STUDY: Participants (N = 126) were accrued through patients who had been treated for prostate cancer at Fox Chase Cancer Center (n = 39) and through newspaper advertisements (n = 87). All participants completed a questionnaire sent by mail. RESULTS: Seventy-four percent of men were probably (50%) or definitely (24%) interested in testing. Participants with a family history of prostate cancer reported that they would be willing to pay substantially more for a genetic test compared with those without a family history. Elevated worry about prostate cancer and concerns about treatment related side effects were associated with greater interest in genetic testing. CLINICAL IMPLICATIONS: Findings demonstrate a need for the development of genetic counseling protocols for at-risk men who are interested in genetic testing, once this test becomes available. PMID- 11898183 TI - High-dose chemotherapy for breast cancer? PMID- 11898184 TI - Resources for caregivers of terminally ill cancer patients. PMID- 11898182 TI - Cancer-related fatigue. Suffering a different fatigue. AB - PURPOSE: The purpose of this naturalistic inquiry was to understand, from the patient's perspective, the differences between cancer-related fatigue (CRF) and "typical" fatigue and to describe its impact on their lives. DESCRIPTION OF STUDY: The sample consisted of 17 adult patients with cancer who agreed to participate, yielding 23 in-depth audio-taped interviews. Participants included inpatients and outpatients from a dedicated cancer center and a Veterans Affairs Hospital. Symbolic interactionism was the framework used for the interviews to understand how individuals and groups construct and discover the meaning of events. Transcribed data were coded by distress category to classify the textual information and to reduce the text to more relevant and manageable bits of data. Five to 10 representative excerpts from each coding category were chosen and were coded independently by a second coder to evaluate intercoder reliability. RESULTS: CRF was found to be more rapid in onset, more energy draining, more intense, longer lasting, more severe, and more unrelenting when compared with "typical" fatigue. CRF caused distress in the physical, social, spiritual, psychological, and cognitive domains of the participants' lives. All participants agreed that CRF was different than the typical fatigue of everyday life. CLINICAL IMPLICATIONS: This study's findings indicate that because of this different fatigue, CRF, participants did not just experience the symptom, but they suffered with it. The current oncology literature and texts do not portray the morbidity of the CRF experience as described by the participants in this study. These data can be used as part of an experiential teaching guide to assist healthcare students and practitioners in understanding the CRF experience. This increased awareness will afford clinicians a better foundation for implementing management strategies. A CRF distress instrument is in development based on these data. PMID- 11898185 TI - Parish nursing--health ministry. PMID- 11898186 TI - A healing lesson through reflexology. PMID- 11898188 TI - At the bedside--patients know the difference. PMID- 11898187 TI - Resources for an environmentally healthy lifestyle. Part II: Clothing. PMID- 11898189 TI - Resources for nurses. PMID- 11898190 TI - The story of Fable: connecting with the real world of being a registered nurse. AB - Fable's story is the result of research using an interpretive approach to explore what it is like to be a registered nurse from the perspective of four new graduates. This story highlights the understanding that nursing is more than just a job: the human connection with patients and staff is a sustaining force during the many tough times experienced by the new grad. PMID- 11898191 TI - My journey into the literature of Therapeutic Touch and Healing Touch: Part 1. AB - This exploration of the literature is in two parts and follows the progress of Healing Touch from a personal perspective. Part one explores Therapeutic Touch and offers a review of current research. Part two in our next issue, will focus on Healing Touch. PMID- 11898192 TI - Utilising paediatric massage in an intensive care unit (PICU) in Saudi Arabia. AB - In this detailed case study of a very sick baby in a modern technological hospital, nursing staff and parents joined forces to implement and monitor a fully coordinated massage program in response to possible ICU psychosis. Positive results indicate that massage enhances the delivery of truly holistic care. PMID- 11898193 TI - Towards a definition of wellness. AB - The term 'wellness' is widely referred to in the nursing literature, yet its meaning remains elusive. Definitions vary widely, if they are provided. Lack of consensus regarding the definition of wellness contributes to fragmentation of access to research findings and limits understanding and action in wellness practice. This paper looks beyond individual definitions of the terms 'wellness' and 'health' to examine some conceptual similarities which may contribute to the debate. PMID- 11898194 TI - A bunch of needles: conflicts and commonalities between East and West. AB - There are fundamental differences between Eastern and Western views of healthcare including the interpretation of 'health' and 'illness', ways in which the human system is perceived, and how diagnoses and treatment are carried out by respective physicians. These concepts are explored in relation to the use of Traditional Chinese Medicines and present nursing theories and practices. PMID- 11898195 TI - Getting on the inside: a researcher's journey into the world of teenage motherhood. AB - This paper describes some of the methodological challenges and particularly the ethical issues when researching with teenage mothers. It specifically describes the difficulties of gaining access, establishing and maintaining relationships and withdrawing from the field. It concludes that close attention to these issues is important when researching with teenage mothers. PMID- 11898196 TI - Self-care tools for creating resistance to burnout: a case study in philophonetics counselling. PMID- 11898197 TI - Understanding what is important for women who live with multiple sclerosis. AB - In this pilot-project eight women with multiple sclerosis (MS), four Continence Nurse Advisers (CNAs) and one researcher formed a participatory action research (PAR) group. Ten group sessions were held in 1997. The CNAs found that 80 of their referrals for continence management were women with MS and they wanted to explore the way in which women manage their incontinence. Although the group was brought together to discuss incontinence, other emergent themes were raised by the women. These themes were: women's ad hoc experience with community services, their problems concerning access to toilets for the disabled, concern with health professionals' lack of knowledge about MS, their formidable experiences of hospitalisation and respite care admissions, their individual efforts in maintaining wellness in the context of chronic illness and the effect of MS on sexual relationships. PMID- 11898199 TI - On being embodied. PMID- 11898198 TI - "Super" presencing nurse healers' stories of healing. AB - This article describes the experience for nurse healers of being a channel for healing. In particular this was explored by revealing "super" presencing--an essential theme of the lived experience of nurse healers, described in a recent research study. "Super" presencing not only enhances our knowledge but gives inspiration in an otherwise undernourished area of the healing profession of nursing. PMID- 11898201 TI - Holistic nursing: the challenge of Jehovah's Witnesses and the issues of blood. AB - This paper explores the issue of blood transfusion from the perspective of a Jehovah's Witness. Deborah discusses the spiritual values underlying the religious commitment of Jehovah's Witnesses and offers many alternatives to blood transfusion. PMID- 11898200 TI - Advancing my health care practice in aromatherapy. AB - This paper describes a clinical trial to examine the effects of aromatherapy on stress reduction in haemodialysis patients. The author used a social readjustment rating scale (SRRS), health assessment questionnaire and reflective journal in the collection and analysis of data. PMID- 11898202 TI - Massaged embodiment of cancer patients. AB - This paper analyses the massaged embodiment of cancer patients and involves a poststructural approach. Poststructuralism as a methodology is used as it provides a way of seeing and understanding cancer patients' complex experience of embodiment. Massage promotes a quality of embodiment that involves well being, comfort and relaxation. However, massage can promote vulnerability and this is evident in some of the participants' talk. For participants, massage provides a reclaiming of 'the old body' and an experience of mind and body connection. Embodiment is theorised as shifting, discursively constituted and multi layered. PMID- 11898203 TI - Suicide first aid. AB - This paper describes some aspects of suicide intervention and outlines a service, and some of the features it offers, to assist in suicide prevention. PMID- 11898204 TI - Dealing with 'post term' pregnancy at home. PMID- 11898205 TI - Migraine in women twenty-six to forty-five years of age. AB - The following research was undertaken within a qualitative framework of analysis. The purpose of this ethnographic study was to explore women's experiences of migraine. Migraine as an illness remains largely misunderstood, misdiagnosed and mistreated. A major theoretical concern is to demonstrate the importance of placing patient's personal accounts of their experiences and understandings at the centre of nursing interest and practice. PMID- 11898206 TI - Ethereal embodiment of cancer patients. AB - Ethereal embodiment is the attending and focusing on the body through discourses such as meditation, visualisation and massage, and the experiencing a new sense of the embodied being as balanced, connected, centred and of being made whole. This paper continues a previous article titled 'Massaged embodiment of cancer patients'. Data from my doctoral studies are analysed utilising crucial concepts of poststructuralism such as subjectivity, discourse, power and history to examine ethereal embodiment. This paper will address the advantages of visualisation and discusses the link between spirituality, embodiment, and memory. PMID- 11898207 TI - Nurse healers: exploring their lived experience as nurses. AB - This article explores the lived experience of nurse healers in nursing, reporting on three themes identified in a recent hermeneutic phenomenological research study. These themes, mapped out in the words of the participants, are termed responding, evolving, and weaving. PMID- 11898208 TI - On the experience of spirituality. PMID- 11898210 TI - Working with the notion of soul in nursing. PMID- 11898209 TI - Eastern and Western paradigms: the holistic nature of traditional Chinese medicine. AB - This paper discusses the practice of holism based upon differing Eastern and Western worldview belief systems. The practice of authentic holistic healing and caring is, according to the author, highly dependent upon which paradigm(s) the nurse lives within, given the constraints of modern Western science and its linear thinking, theoretical laws and objectivity towards the practice of healing and health care. From a Traditional Chinese Medicine perspective, healing and health are based upon circular thinking, observation, subjectivity and feeling, which reflects a differing holistic approach to human beings. PMID- 11898211 TI - Holistic care and spirituality: potential for increasing spiritual dimensions of nursing. AB - Holistic care addresses the physical, psychological, social and spiritual dimensions of the client. Health care providers, however, frequently overlook the spiritual dimension. Spiritual care is inseparable from physical, social and psychological care because together they form the whole. Promoting spiritual well being supports clients in their journey to find meaning and hope in life and peace in death. The preparation of nurses for the spiritual aspects of care deserves much more attention. PMID- 11898213 TI - A changed life. PMID- 11898212 TI - Light relief: the case for ocular light therapy. AB - Ocular Light Therapy is emerging as a significant form of therapy, particularly in psychological and psychosomatic disorders. OLT involves the projection of light through colour filters to the eyes of the client, with consequent photocurrent stimulation of cortical and hypothalamic systems. PMID- 11898214 TI - Cancer resolution or misdiagnosis? PMID- 11898215 TI - Experience awareness tools for preventing burnout in nurses. AB - Philophonetics counselling is a radically new method of coping with stress through the development of individually tailored self-controlling emotional and cognitive strategies. This paper proposes that philophonetics counselling provides an innovative holistic approach of dealing with burnout in the nursing profession. PMID- 11898216 TI - Combining complementary therapies and nursing skills to supplement healing. AB - The biomedical model's main focus is 'cure' which sometimes is not the most important outcome as far as clients are concerned. The need for other techniques to supplement and aid 'healing' rather than 'cure' can have positive outcomes for both the nurse as healer and the client. PMID- 11898217 TI - Empowering female residents: alternative approaches to urinary tract infections (UTIs) in nursing homes. AB - The value of practical and literary nursing research is compared to the rationale that underlies orthodox and alternative treatment management for UTIs. A new policy for treating UTIs is described and proposed, with consideration given to the impact of this policy upon nursing home residents and staff. PMID- 11898218 TI - On fixing stress from the inside and outside. PMID- 11898219 TI - The complex 'whole': exploring homoeopathic and spiritual considerations. AB - Unlike the biomedical model, holistic health care takes a much broader view of what constitutes health and the responsibility for helping restore an individual's health. Homoeopathy addresses the physical, mental and emotional aspects of the whole individual, while alternative practices such as yoga, hypnotherapy and meditation can be described as 'functional' spiritual practices which demonstrate the taking of personal responsibility for health care to the individual. PMID- 11898220 TI - Guided imagery: a strategy for improving relationships and human interaction. AB - Guided imagery, rather than being a revolutionary complementary or alternative therapy, has been utilised for centuries to alleviate suffering and promote wellness. Its efficacy has been studied and documented in physical, psychological and spiritual dis-ease. This article reviews relevant literature supporting the premise that guided imagery is a strategy that can be used for improving therapeutic relationships and is an example of holistic health care. PMID- 11898221 TI - Via negativa: considering caring by way of non-caring. PMID- 11898222 TI - Ocular light therapy: a case study. PMID- 11898223 TI - Traditional Chinese medicine: a case of dysmenorrhoea. AB - Acupuncture treatment continued for three cycles, undertaken one week prior to menses whereas herbal therapy was maintained for six months. Within this six month time frame Jenny's overall health status had changed dramatically for the better. She was pain free and consequently her life-style had improved to the point where she felt confident to travel overseas for a holiday. To date, Jenny and her mother have kept in touch with clinic staff and no further treatment has been necessary. PMID- 11898224 TI - Mere male cares. PMID- 11898225 TI - Picking up the pieces. AB - This paper examines some of the current evidence of stress and burnout among nurses and other health care workers. Even in environments where pay and conditions are good, nurses still experience significant difficulties in healing relationships, stress and burnout sufficient to render them dysfunctional at work. The paper, using the case study of a unit set up to provide care for nurses, examines how the needs of nurses can also be taken into account in the holistic perspective on caring. PMID- 11898231 TI - The cancer agenda and the Y2K elections. PMID- 11898232 TI - Screening to identify and treat tamoxifen-associated side effects. PMID- 11898233 TI - Patients' sensations after breast cancer surgery. A pilot study. AB - PURPOSE: The purpose of this study was to obtain information about the prevalence and characteristics of breast sensations after breast cancer surgery, the impact they had on patients, and aggravating and relieving factors. DESCRIPTION OF STUDY: Within 1 month after the date of their surgery, 132 patients with breast cancer completed an instrument rating the prevalence, severity, and level of distress of breast sensations. Information also was obtained on the impact that those sensations had on activities of daily living and factors that triggered and provided relief from sensations. RESULTS: Certain sensations remain prevalent (numb, tender), severe (burning, sharp), and distressing (cramping, painful). Overall these sensations significantly interfered with patients' activities of daily living. Certain activities (movement, position change) triggered sensations, while others (position change, medication) provided relief. CLINICAL IMPLICATIONS: Healthcare professionals can use the information learned from this pilot study to educate patients both preoperatively and postoperatively about prevalent breast sensations after surgery for breast cancer and about the types of activities that may aggravate or relieve these sensations. As patient educators and advocates, oncology professionals must continue to explore the long term effects and treatment options to provide optimal care and support to patients who have or are likely to have post-surgical breast sensations. More studies are needed to explore long-term effects and treatment options. PMID- 11898234 TI - Smoking behavior, knowledge, and beliefs among Korean Americans. AB - PURPOSE: The purpose of the study was to examine smoking behavior, knowledge, and beliefs among Korean Americans. DESCRIPTION OF STUDY: One hundred four Korean American men and 159 women, 40 to 69 years old, living in Chicago, Illinois, served as study respondents. The National Health Interview Survey (NHIS) Cancer Control Supplement Questionnaire was used to collect data. The NHIS was translated into Korean with minor modifications to develop a culturally appropriate instrument. RESULTS: This study suggests that smoking is almost exclusively a male behavior (38.5%) and that Korean American men smoke mostly cigarettes. Almost 90% of women have never smoked, whereas 23% of men reported never smoking. Respondents with a non-Christian background or no religious affiliation were 16.5 times more likely to be current smokers. Respondents who had lived in the United States less than 10 years were 12.5 times more likely to be current smokers. More than 90% of men, regardless of smoking status, were able to identify an association between smoking and major chronic diseases. CLINICAL IMPLICATIONS: The prevalence of smoking among these Korean American men places them at considerable risk for smoking-related disease. Healthcare providers must be better informed about smoking behavior in this group, and specific attention to recently migrated men and those reporting religions other than Christianity is recommended. Health-protecting strategies for women and children who fall victim to secondhand smoke, or who may be targeted by tobacco advertising, are also an important step in disease prevention for this population. PMID- 11898235 TI - Exercise and weight gain in breast cancer patients receiving chemotherapy. AB - PURPOSE: Weight gain is a common side effect for women receiving adjuvant chemotherapy and may have negative long-term implications for survival. The purpose of this study was to examine the effects of aerobic exercise on weight gain in women with breast cancer receiving adjuvant chemotherapy. DESCRIPTION OF STUDY: Seventy-eight women who had recently received a diagnosis of breast cancer and who were beginning adjuvant chemotherapy were enrolled in a home-based exercise study during the first four cycles of chemotherapy. Weight change, body mass index, anorexia, nausea, fatigue, caloric expenditure during exercise, and functional ability were recorded. RESULTS: Women who adhered to the exercise program maintained their body weight, while nonexercisers steadily gained weight (P < .05). There were no differences in incidence or intensity of nausea or anorexia between the exercisers and nonexercisers. Women who exercised over the four cycles of chemotherapy improved their functional ability (mean 23%) compared to the nonexercisers who showed significant declines in functional ability (mean 15%). CLINICAL IMPLICATIONS: Exercise may be an effective intervention to minimize weight gain in women with breast cancer who are receiving adjuvant chemotherapy. Preventing weight gain in these patients may be important in preventing recurrent disease and other comorbidities associated with excess weight. PMID- 11898236 TI - Cancer survivor identity and quality of life. AB - PURPOSE: The purpose of this article is to propose a framework for enhancing an understanding of quality of life among cancer survivors that takes into account individuals' subjective experiences and attributions of self as they relate to their own experience with cancer. OVERVIEW: This review of a small yet burgeoning cancer survivorship literature posits that there is room to expand the theoretical context for understanding cancer and its impact on quality of life by assuming that the experience of cancer involves changes in social roles and identity. In turn, changes in the way cancer survivors see themselves in relation to the world, including the ability to carry out roles and responsibilities, may affect quality of life. This perspective suggests that a cancer diagnosis initiates a survival trajectory and a social role that extends over the remainder of one's life, regardless of life expectancy. CLINICAL IMPLICATIONS: Expanding the understanding of quality of life and the factors that contribute to it should help subsequent investigations of who might benefit from which psychosocial support interventions provided when (i.e., during early diagnostic stage, treatment stage, or both or in the long term). Equipped with this understanding, clinicians and agencies providing services to cancer survivors and their families can develop appropriate supportive interventions that facilitate and enhance quality of life. PMID- 11898237 TI - Complementary/alternative therapies. Potential safety issues in cancer care. PMID- 11898238 TI - Resources for treatment of tobacco dependency. PMID- 11898239 TI - Targeting leukemia cells with gemtuzumab ozogamicin. PMID- 11898240 TI - Integrating outcomes into computerized information systems. It's time to get guidelines off the shelf. AB - The tools used to measure outcomes in the 21st century probably will look nothing like the tools used to measure outcomes during the past century. These guideposts (guidelines, information systems, and vocabulary) are only three of the many new tools that will be available to us through technology. The three guideposts described in this column are necessary ingredients for success in managing outcomes and demonstrating accountability. They look like the lampposts to a bright future. PMID- 11898241 TI - Supporting nurses in their quest for evidence-based practice. Research utilization and conduct. PMID- 11898242 TI - Physical and emotional patient safety. A different look at nursing-sensitive outcomes. AB - Data on nursing-sensitive outcomes, beyond traditional isolated indicators such as pressure ulcers, are often unavailable for nurses to evaluate the overall quality of their care. This article describes a quality improvement effort to provide nurses with both a positive and a negative patient outcomes score. The approach can make visible the often invisible role of nurses in the growing field of patient safety. PMID- 11898244 TI - Effectiveness of different medication management approaches on elders' medication adherence. AB - This pilot study investigated the effects of three different medication management approaches on medication adherence and resource utilization. Sixty-one participants living in an independent elder community in South Florida were randomly assigned to one of the three medication management approaches: (1) a pillbox method, (2) a voice-activated method, and (3) self-administration of medications as they had in the past. One outcome was measured by recording the number of doses of medications ingested over a 1-, 3-, and 6-month period. Adherence to medications also was measured by the impact on the medical diagnosis. For example, the hypertensive group was defined adherent by a sustained normotensive pressure. Participants' medical records were examined as to the number of physician office visits, hospitalizations, and home health visits. There were significant differences in the mean number of doses missed, with the fewest in the voice-activated group to the highest in the self administration (control) group. Additionally, the group that self-administered their own medications had more frequent physician office visits and increased hospitalizations. Because the elder population is prone to medication mismanagement for a variety of reasons, nurses are in a unique position to identify populations at risk and suggest interventions that may improve medication adherence. PMID- 11898243 TI - Prescreening cardiothoracic surgical patient population for post acute care services. AB - For the postoperative cardiothoracic patient, the first 2 weeks during recovery is the most difficult time. Initiating discharge planning at the earliest opportunity aids in identifying those patients at risk for functional status disability. The purpose of this study was to examine the results of a pilot preadmission screening program to determine whether cardiothoracic surgical patients could be accurately identified for postoperative needs during preadmission testing. PMID- 11898245 TI - Focus groups. Giving voice to people. AB - The authors used focus groups to evaluate the effectiveness of specific patient teaching tools on patient education outcomes. In addition, data from the focus groups identified both deficiencies in the teaching-education process and ways to improve patient education and self-care outcomes. Negative and positive aspects of using focus groups in outcome measurement are included. PMID- 11898246 TI - Beyond interviewing skills: twelve steps for training interviewers. AB - The measurement of many outcomes relies on patients and their families for information that can best be collected through interviews. The authors describe 12 practical suggestions that help ensure achievement of subject enrollment and interview goals. PMID- 11898252 TI - Confronting unfounded fears in cancer care . PMID- 11898253 TI - A steroid-induced disorder in a patient with non-Hodgkin's lymphoma. PMID- 11898254 TI - Partner abandonment of women with breast cancer. Myth or reality? AB - PURPOSE: The purpose of this article is to determine the existing evidence related to marital breakdown after a breast cancer diagnosis by reviewing studies that highlight two current belief models: the lay belief model and the clinical belief model. OVERVIEW: The small number of studies conducted on this topic since 1988 revealed no data to confirm the lay belief model, which proposes that women with breast cancer are abandoned by their partners. The evidence appears to support the clinical belief model that the majority of marital relationships remain stable after breast cancer and that breakdown is most likely in those relationships with pre-existing difficulties. CLINICAL IMPLICATIONS: This review indicates that it may be important for clinicians to routinely ask about the quality of the marital relationship as part of the initial assessment, because it appears that this may be a main predictor of post-diagnosis marital adjustment. In addition, greater dissemination of the findings of this review through the media and through cancer organizations is needed to more accurately reflect the experience of couples facing breast cancer and, thus, to begin to change the public perception of partner desertion after breast cancer. This could help both women with breast cancer and women from the general population who may one day confront a breast cancer diagnosis. PMID- 11898255 TI - Early postsurgery experience of prostate cancer patients and spouses. AB - PURPOSE: The authors describe the experience of men with prostate cancer and their spouses in the early recovery period after surgery. DESCRIPTION OF STUDY: As part of a longitudinal qualitative study, semistructured interviews were held with 34 patients who had prostate cancer and their spouses 8 to 10 weeks after surgery. RESULTS: Five components of experience emerged from the interviews: 1) hearing news about the extent of their cancer after surgery influenced how patients viewed their cancer experience and, in many cases, their recovery; 2) men placed great emphasis on recovering their physical capacity quickly; 3) couples connected with each other through working out care routines and managing periods of irritability; 4) couples described a range of responses to surgery side effects and complications; and 5) the meaning of cancer varied for couples, with most seeing the experience as a temporary disruption. CLINICAL IMPLICATIONS: Physicians, nurses, social workers, and other health professionals working with patients before and after prostatectomies may assist couples to prepare better for the early recovery period by being both sensitive to the men's need to recover physical capacity quickly while helping them to understand that recovery takes time. Accurate information about expected periods of irritability, side effects, and possible complications would diminish the likelihood of distress during this period. PMID- 11898256 TI - Delirium in patients with cancer at the end of life. AB - PURPOSE: Delirium is a common and distressing syndrome seen in patients with advanced cancer. Behavioral manifestations of delirium, such as agitation, may result in medical intervention, stress to family caregivers, and inpatient hospice admission. The purpose of this study was to examine the frequency, characteristics, and presumed causes of delirium in patients with advanced cancer. DESCRIPTION OF STUDY: Records of all patients with cancer who were admitted to an inpatient hospice facility in 1995 were reviewed retrospectively (N = 210). Patients were classified as delirious based on the clinical judgment of the admitting physician. RESULTS: Delirium was the third most common reason for admission (20%). Male gender (P = .04) and the presence of a primary or metastatic brain tumor (P = .03) were significant risk factors for delirium, while advanced age and primary or metastatic liver, lung, or bone cancer were not. Resolution of the agitation, the most disruptive symptom of delirium, occurred in 69% of patients before death or discharge. CLINICAL IMPLICATIONS: Delirium is common in hospice patients with cancer and is an important cause of family distress and increased cost of care. The recognition of early clinical signs and predisposing factors should facilitate prompt diagnosis. Appropriate intervention is usually successful in alleviating the most distressing symptoms of delirium. PMID- 11898257 TI - Attitudes toward genetic testing in patients with colorectal cancer. AB - PURPOSE: The purpose of this study was to examine risk perceptions and interest in genetic testing among African American and White patients with colorectal cancer. DESCRIPTION OF STUDY: In this cross-sectional study, 98 patients with colorectal cancer participated in a brief structured telephone interview. Information was collected on knowledge and risk perceptions regarding colorectal cancer genetics, health behaviors, knowledge about the availability of genetic testing, and interest in genetic testing for colorectal cancer susceptibility. RESULTS: Sixty-one percent of the participants were worried about their relatives' risk of colorectal cancer, and 64% were concerned about being a colorectal cancer susceptibility gene carrier. Although 81% of the participants reported that they had never heard about a genetic test for colorectal cancer susceptibility, 72% stated that they would want to take the test if it were available. Predictors of intention to have a genetic test were younger age, less advanced stage of disease, and more frequent thoughts about colorectal cancer being hereditary. CLINICAL IMPLICATIONS: These results suggest that the demand for genetic testing may be great, despite a general lack of knowledge about colorectal cancer genetics and the potential risks and limitations of testing. Education and counseling about cancer genetics and genetic testing may clarify misconceptions about hereditary colorectal cancer and help patients with colorectal cancer and their family members make informed decisions about whether to undergo testing. PMID- 11898259 TI - Sentinel lymphadenectomy for breast cancer. A standard of surgical care? PMID- 11898258 TI - Developing a community program on cancer pain and fatigue. AB - PURPOSE: The overall purpose of this project was to establish a community-based educational model on pain and fatigue management for individuals with cancer. The specific aims were: 1) to develop an appropriate educational program; 2) to pilot test this program in a community setting that supported a self-care approach; and 3) to evaluate the program process and outcomes. DESCRIPTION OF PROGRAM: The I Feel Better program was implemented through a two-session educational workshop taught by masters-prepared oncology nurses and was held at four Southern California sites of The Wellness Community. The focus of the sessions was to provide participants with general information about each symptom, assessment and management of those symptoms, and strategies for effectively communicating with their healthcare providers. Sessions of 2.5-hour duration were held on Saturday mornings and required preregistration. RESULTS: The participants were primarily female and White, with an average age of 58 years. Participants reported considerable pain and fatigue. They also lacked accurate information about pain management. Program evaluation revealed that the content and format were well received by the participants. They rated the program as extremely useful and reported positive outcomes after the first session. CLINICAL IMPLICATIONS: This pilot educational intervention program has strong implications for multidisciplinary educational approaches for patients with cancer. Limitations resulted from the setting selected and the possibility that participants were already active in their fight against cancer. Generalization to other community settings may not be as successful. Programs could be cosponsored by several collaborating institutions to share resources. Referral to community programs by physicians, nurses, and social workers can occur as needed when identified during patient interventions. The voluntary participation of health professionals in community education programs could provide a valuable service for patients and a rewarding experience for educators. PMID- 11898260 TI - Information sources. Cancer genetics and genetic testing. PMID- 11898262 TI - Loss prevention case of the month. What really is the "standard of care"? PMID- 11898263 TI - Cultural models of long-term care. AB - Long-term care remains a formidable challenge in the spectrum of geriatric services, influenced by cultural attitudes, funding priorities, societal needs, and personal preferences. Many positive components exist in American models of long-term care, including medical directorships, mid-level practitioners, and regulatory control, however home and community-based services are relatively under-developed compared to the experiences in other countries. PMID- 11898261 TI - Oxaliplatin. A platinum compound for colorectal cancer? PMID- 11898264 TI - Clostridium difficile infection associated with levofloxacin treatment. AB - Nine cases of Clostridium difficile (CD) infection were observed in the period of six months at a nursing home. Eight of them occurred during or after antibiotic treatment. Levofloxacin was used alone in three cases and in combination with another antibiotic in three other cases. CD infection occurred with other antibiotics in two cases. In one case, CD infection occurred without any antibiotic treatment. It is generally accepted that quinolones rarely cause CD infection. Levofloxacin, a new antibiotic of a quinolone group, appears to be an exception. Considering the endemic level of CD infection and the high mortality and morbidity among elderly residents in the long-term care facilities, CD infection should be considered one of the major adverse effects of antibiotic therapy. Physicians are cautioned to prescribe antibiotics judiciously and to anticipate CD infection during and after antibiotic treatment. PMID- 11898265 TI - The insanity defense: Tennessee's process and outcome. PMID- 11898266 TI - Health's role with anthrax. PMID- 11898267 TI - Stem cell research. PMID- 11898268 TI - The continuing crisis. PMID- 11898269 TI - Somebody help me, please. PMID- 11898270 TI - W. D. Miller. The pioneer who laid the foundation for modern dental research. AB - Toothache has been the most ubiquitous ailment to plague mankind from time immemorial. Until the late 1700s, it was thought that the cause of this torment was the wriggling, in a carious tooth, of a worm. And early attempts at treatment were focused on driving the "worm" out. It was one of the world's greatest scientists, the dentist W. D. Miller, who, after extensive research, in 1891 published his epochal work, The Microorganisms of the Human Mouth, which set forth a new theory regarding the cause of dental caries. His postulating a "chemico-parasitic" origin of caries laid the basis for all the modern research in dentistry aimed at wiping this scourge out. Contemporary research has proven the worth of Miller's groundbreaking theory. PMID- 11898271 TI - Dental caries. Diagnosis and treatment. AB - The diagnosis of tooth decay must go beyond the clinical detection of a carious lesion. The practitioner should assess the individual's risk factors as well as the activity of the lesion. Our traditional instrumentation has limitations. Therefore, researchers are urged to find new diagnostic tools to allow earlier detection, to predict disease activity and, finally, to assess the susceptibility of an individual. PMID- 11898273 TI - An expression of depression. Help end the suffering by recognizing the signs. PMID- 11898272 TI - Spontaneous angioedema of oral cavity after dental impressions. AB - Maxillofacial angioedema is a rare condition encountered by the oral and maxillofacial surgeon. Its significance lies in its potential to partially or totally obstruct the upper airway secondary to acute sudden swelling. In some individuals, angioedema is hereditary; in others, it occurs spontaneously, without warning, as an allergic reaction. The case presented here documents perioral angioedema secondary to dental impressions and reviews its management. PMID- 11898274 TI - DASH diet, reduced salt lowers blood pressure. PMID- 11898276 TI - Few eligible stroke patients receive lifesaving treatments. PMID- 11898275 TI - Protecting kids from medical errors. PMID- 11898277 TI - Research protections expanded; new guidelines forthcoming. PMID- 11898278 TI - Surgeon general's oral health report finds profound disparities. PMID- 11898279 TI - New pediatric growth charts target weight problems. PMID- 11898280 TI - Viral changes causing chronic hepatitis C. PMID- 11898281 TI - Coming to consensus on osteoporosis. PMID- 11898282 TI - Reporting adverse events and product problems. MedWatch provides comprehensive resource for health professionals. PMID- 11898283 TI - Preventing preterm birth. Evidence-based interventions shift toward prevention. PMID- 11898284 TI - Battered woman in a small town. Encountering violence through the story of one woman. PMID- 11898285 TI - Working toward cultural competence. Making the first steps through cultural assessment. PMID- 11898286 TI - Teaching with toys. Invigorating your style with creativity. PMID- 11898287 TI - The river running through Us. Exploring the implications of nursing education. PMID- 11898288 TI - Where are the nurses? PMID- 11898289 TI - The development of clinical practice guidelines for the use of aromatherapy in a cancer setting. AB - The need to develop guidelines for the use of aromatherapy was identified to ensure safe and appropriate use in clinical practice. Therapeutic actions and safety data were reviewed along with literature relating to nurses' use of aromatherapy. A policy was developed with the needs of the oncology patient being the foremost consideration. Methods of administration and dosage of essential oils were stipulated to address safety issues. PMID- 11898290 TI - Generating theory from the client's experience of same day laparoscopic sterilisation. AB - A grounded theory methodology was used to generate theory about the experiences of women undergoing laparoscopic sterilisation in a day surgery unit. Theories were developed on the role of client education in relieving anxiety, and the importance of privacy in all stages of client care. The grounded theory approach offered a client-centered model for day surgery planning. PMID- 11898291 TI - My journey into the literature of Therapeutic Touch and Healing Touch: Part 2. AB - Following on from part one published in the previous issue, research on Healing Touch will be now be reviewed and applied to current nursing theory to support this exciting holistic modality. PMID- 11898292 TI - Using Bowen Therapy to improve staff health. AB - Positive results from the administration of Bowen Therapy to staff while at work has prompted an innovative project addressing the lowering of stress levels and preventing burn-out for all staff, in and beyond nursing. PMID- 11898293 TI - Tai Chi: towards an exercise program for the older person. AB - In the changing demographic and health care contexts of aged care, the creation of health promoting exercise programs for frail elderly people to improve wellbeing and quality of care is a common objective in community and residential aged care. This article describes the background to the development of a gentle exercise program that has therapeutic value and the potential to improve wellbeing and quality of care. PMID- 11898294 TI - Cultural borrowing and sharing: aboriginal bush medicine in practice. AB - Aboriginal people of the Northern Territory continue to approach health related issues by integrating traditional and modern lifestyles and sharing information. PMID- 11898295 TI - Working with dementia patients and their relatives. PMID- 11898296 TI - Saying no to circumcision: ending cycles of abuse. PMID- 11898297 TI - Outcome-based research: Part II. PMID- 11898299 TI - Whose fault? PMID- 11898298 TI - Aromatherapy for Health Professionals annual report. PMID- 11898300 TI - Implementation of a change process to improve outcomes of patients admitted to epilepsy monitoring unit. AB - A continuous quality improvement in health care, combined with measurement of patient and organizational outcomes, is an international trend that promises improvement in meeting the needs of health care consumers in a cost-effective manner. A clinical improvement workshop was conducted in our institution to study the practical methods that can be used by clinical teams to improve the quality and value of health care. This workshop enabled the Epilepsy Team to identify change strategies that potentially could accelerate clinical improvement efforts in the epilepsy monitoring unit (EMU). The Team was able to develop, plan, and test improvements and link these improvements to patient outcomes. PMID- 11898301 TI - Nurses' narratives of outcomes after delegation to unlicensed assistive personnel. AB - Although unlicensed assistive personnel (UAP) have been a part of health care teams for decades, today's UAP are assisting in the care of more acutely ill clients who are being discharged after shorter hospital stays. This qualitative study examined nurses' narratives of patient outcomes after delegation of activities to UAP and identified the factors leading to the outcomes. PMID- 11898302 TI - Beyond the acute care setting: community-based nonacute care nursing-sensitive indicators. AB - In 1994, concerns about the effects of hospital restructuring on patient care resulted in the American Nurses Association (ANA) undertaking a major, long-term initiative. Nursing's Safety & Quality Initiative (the Initiative) was designed to measure the impact of such changes on patient care. The Initiative has three major foci: research, continuing education, and legislation/policy. This article addresses a recent development in the research component of the Initiative, involving the identification of nursing-sensitive indicators for community-based nonacute care. PMID- 11898303 TI - Measuring behavioral and mood disruptions in nursing home residents using the Minimum Data Set. AB - The Minimum Data Set (MDS) is a standardized assessment tool designed to provide a comprehensive biopsychosocial assessment of medical, behavioral, and cognitive status of nursing home residents. This pilot study examined the relationships of three MDS subscales--cognition, depressive symptoms, and behavioral disruptions- to other measures of the same domains (e.g., diagnosed dementia and depression and caregiver ratings on the Revised Memory and Behavior Problems Checklist [RMBPC]). The sample consisted of 135 nursing home residents with a mean age of 84 years. Based on the MDS, there was a high prevalence of cognition-related behaviors but a low prevalence of disruptive and depressed behaviors. The prevalence rates were substantially different according to the RMBPC. In addition, most of the MDS subscales failed to differentiate between residents with and without diagnosed dementia and depression, whereas caregiver ratings on the RMBPC did. The MDS and RMBPC subscales were modestly related but only in residents without dementia. These findings raise questions about the validity of the MDS in measuring nursing home residents' behavior, especially depressive and disruptive behaviors. Thus, caution should be employed in using the MDS as a sole outcome measure for these behaviors, and the use of multiple measures is suggested. PMID- 11898304 TI - Care management. PMID- 11898305 TI - Using patient acuity data to manage patient care outcomes and patient care costs. AB - This article describes actual reported uses for patient acuity data that go beyond historical uses in determining staffing allocations. These expanded uses include managing patient care outcomes and health care costs. The article offers the patient care executive examples of how objective, valid, and reliable data are used to drive approaches to effectively influence decision making in an increasingly competitive health care environment. PMID- 11898306 TI - Pain outcomes after intestinal surgery. AB - Pain sensation and distress in 38 intestinal surgical patients were moderate to severe on postoperative day 1, ranging from 34 to 49 mm and 33 to 45 mm, respectively, on 100-mm scales. During ambulation, both increased from baseline to post-ambulation, P < 0.01. Half of the patients reported severe pain not relieved by analgesics, and although 44% learned a relaxation technique in the past, only 8% used one for pain after this surgery. Pain disturbed the sleep of 34% of the patients, and pain was related to respiratory, intestinal, febrile, and other complications in 18 (47%) subjects. Attentive analgesic use and nonpharmacologic therapies are recommended. PMID- 11898307 TI - Methodological considerations: using large data sets. AB - Studies in the field of outcomes research often use very large data sets. Before data collection, specific issues related to data storage, data collection, and data retrieval should receive careful consideration. New technologies have made the process more cost-effective, efficient, and under the direct control of the researcher. PMID- 11898308 TI - To err is human. PMID- 11898309 TI - Health outcomes skills for care management. AB - There are several skills nurses can develop or expand to improve their ability to manage outcomes. This is an exciting area for nurses and one that is valued during times of cost reduction in healthcare. In addition to increasing skills in the health outcomes management arena, nurses will benefit from a continual increase in knowledge of case management, clinical pathways, and guidelines. The healthcare market-place is likely to reward nurses with these skills. PMID- 11898310 TI - Methodological challenges in the study of hospital readmissions. AB - Throughout the literature on hospital readmission, little has been written about what is an optimal rate of rehospitalization and the definition of preventable hospitalization. Therefore, the research on hospital readmissions aids in the understanding of variation in the phenomenon but does not necessarily set goals or standards of practice. Using secondary data rather than primary data does not preclude careful thought regarding the operationalization of each variable under study. The optimal time to initiate that process is at the early conceptual phase of a project. As researchers and clinicians proceed with planning a readmission study, reviewing these six categories of methodological issues will assist in producing a carefully designed definition of readmission. PMID- 11898311 TI - Use of variance outcomes to improve the management of the adult kidney transplant patient. AB - Implementation of clinical pathways requires the measurement of outcomes to foster ongoing improvement in patient care. The use of variance information can optimize patient outcomes and enhance and refine clinical practice. This article discusses the use of the variance outcomes of a clinical pathway to improve the care of adult post-kidney transplant patients. PMID- 11898312 TI - Simplified scoring and psychometrics of the revised 12-item Short-Form Health Survey. AB - The 12-item Short-Form Health Survey was developed to describe mental and physical health status of adults and to measure the outcomes of healthcare services. Based on testing of the original SF-12 with a group of older adults, a revised scoring system and measurement model of the SF-12 Health Survey was proposed. The purpose of this study was to test the reliability and validity of this revised measurement model and scoring system. Testing was done with a sample of 187 older adults in a continuing care retirement community and a sample of 211 older adults discharged from an acute care setting. There was sufficient evidence for the internal consistency of the revised SF-12 (Cronbach alpha coefficients of 0.72 to 0.89); test retest reliability (r = 0.73-0.86); reliability based on R2 values; and validity based on confirmatory factor analysis, contrasted groups, and hypothesis testing. The revised SF-12 is a valid and reliable measure that can be used with confidence to measure outcomes for older adults. PMID- 11898313 TI - Using OASIS patient outcomes to evaluate a cardiac disease management program: a case study. AB - The purposes of this article are to present a case study that demonstrates the use of OASIS data in evaluating a cardiac disease management program and to identify the problems encountered and the knowledge gained. It was found that OASIS data can be useful in the description of patients in disease management program development. The analysis of patient end-result outcomes (comparing start of care and discharge information) proved to be the greatest challenge. Recommendations for future studies are included. PMID- 11898314 TI - Challenge of managing interventional cardiology patients over a decade. AB - Using an outcomes management approach, St. Luke's Episcopal Hospital has achieved positive outcomes in the interventional cardiology patient population. In this article the authors describe a decade of experiences related to the processes they implemented and the outcomes achieved in this patient population. PMID- 11898315 TI - Work site disease management outcomes: expanding the role of the APN. AB - Outcomes of a work site disease management program managed by an advanced practice nurse were evaluated. Fifty-four participants were surveyed. A significant number of participants with dyslipidemia and diabetes reported that the program positively changed individual health behaviors. Those with dyslipidemia and hypertension reported improved understanding of their condition after being involved with the program, and participants with asthma indicated more control of their condition. Program satisfaction was high. Continued development and implementation of work site programs are indicated to improve health outcomes of employees. PMID- 11898316 TI - Environmentally safe places for children. PMID- 11898318 TI - Feeling more at home with homeopathy. PMID- 11898319 TI - Incorporating holism into patient care. PMID- 11898320 TI - Educating our state boards of nursing on holistic nursing practice--Part I. PMID- 11898321 TI - Oddenino's consciousness and the art of internal spiritual healing. PMID- 11898323 TI - The White House Commission on Complementary and Alternative Medicine Policy: Flexner Report II? PMID- 11898324 TI - Update on the standards for advanced holistic nursing practice. PMID- 11898325 TI - The ties that bind. PMID- 11898326 TI - Nurse case management skills required for care management. PMID- 11898327 TI - Achieving outcomes in a joint-appointment role. PMID- 11898328 TI - Effects of individual performance feedback on nurses' adherence to pain management clinical guidelines. AB - The purpose of this study was to examine the effects of an individual performance feedback intervention on nurses' adherence to pain management practice guidelines. Thirty orthopedic staff nurses received individual performance feedback on their past performance of three recommended pain management practices. Nurses' performance of the three recommended practices significantly improved over the 15 weeks after the feedback intervention. Missed 4-hour pain assessments declined (t = 8.77, df = 239, P < 0.0001), reassessments after analgesics increased (t = -5.71, df = 239, P < 0.0001), and follow-up taken for unacceptable pain increased (t = -3.08, df = 178, P < 0.01). Results suggest that individual performance feedback may be an effective approach to improving nurses' pain management intervention activities. PMID- 11898329 TI - Comparison of home health care outcomes and service use for patients with wound/skin diagnoses. AB - The purpose of this comparative study was to evaluate patient outcomes for wound healing and home health care service use for patients whose care was paid for by the traditional Medicare program versus a Medicare managed care organization (MCO). Results showed that there were no differences between the groups in wound healing at discharge, functional ability at discharge, or the numbers of home visits. Changes in the Medicare reimbursement system for home health care may have provided an equalizing effect between Medicare MCO and traditional Medicare patients with wound/skin diagnoses in home health care agencies in the Midwest. PMID- 11898330 TI - Benchmarking: measuring the outcomes of evidence-based practice. AB - Measurement of the outcomes associated with implementation of evidence-based practice changes is becoming increasingly emphasized by multiple health care disciplines. A final step to the process of implementing and sustaining evidence supported practice changes is that of outcomes evaluation and monitoring. The comparison of outcomes to internal and external measures is known as benchmarking. This article discusses evidence-based practice, provides an overview of outcomes evaluation, and describes the process of benchmarking to improve practice. A case study is used to illustrate this concept. PMID- 11898331 TI - Community Health Nursing Outcomes Inventory. AB - The Community Health Nursing Outcomes Inventory (CHNOI) is a 48-item assessment instrument that measures client and nurse outcomes in community settings. The objective of this study was to test the psychometric properties and feasibility of the CHNOI. Results suggest that the CHNOI is an instrument that efficiently measures outcomes sensitive to nursing care in health services research. Managers and researchers may find this instrument useful to assess a community health program's effectiveness. PMID- 11898332 TI - Structured and nonstructured exercise in a corporate wellness program. A comparison of physiological outcomes. AB - The devastating effects from cardiovascular disease are the largest contributors to employers' health care costs, insurance premiums, disability insurance, and worker's compensation. The purpose of this study was to establish baseline data regarding physiological outcomes comparing two participant groups in a corporate wellness program. Results suggest that a corporate wellness program can be beneficial in assisting employees to improve their health behaviors and outcomes. PMID- 11898333 TI - Determining the relationship between end-of-life decisions expressed in advance directives and resuscitation efforts during cardiopulmonary resuscitation. AB - In recent years, there has been an increased focus on end-of-life decisions and the use of medical technology. It is not well documented in the literature whether or not and to what extent patients' advance directives are used for directing resuscitative efforts. The purpose of this study was to determine how useful patients' advance directives were to members of the health care team in determining treatment and end-of-life decisions among patients who received cardiopulmonary resuscitation (CPR) efforts. Medical records of 135 adult patients who had undergone CPR efforts within the previous year were reviewed to determine if and to what extent advance directives were useful in directing end of-life care and treatment decisions. Only 35 of these patients had advance directives. Three categories for advance directives emerged: those that were "independently directive," those that were "vague and required further clarification," and those that were "nondirective." Information from this study may be used to clarify treatment options for end-of-life care and to determine if and what further interventions are required to ensure that advance directives can be executed as meaningful documents. PMID- 11898334 TI - Biochemical confirmation and characterization of the family-57-like alpha-amylase of Methanococcus jannaschii. AB - The gene encoding a family-57-like alpha-amylase in the hyperthermophilic archaeon Methanococcus jannaschii, has been cloned into Escherichia coli. Extremely thermoactive alpha-amylase was confirmed in partially purified enzyme solution of the recombinant culture. This enzyme activity had a temperature optimum of 120 degrees C and a pH optimum 5.0-8.0. The amylase activity is extremely stable against denaturants. Hydrolysis of large sugar polymers with alpha-1-6 and alpha-1-4 linkages yields products including glucose polymers of 1 7 units. Highest activity is exhibited on amylose. The catalyst exhibited a half life of 50 h at 100 degrees C, among the highest reported thermostabilities of natural amylases. PMID- 11898335 TI - Novel, thermostable family-13-like glycoside hydrolase from Methanococcus jannaschii. AB - A novel glycoside hydrolase from the hyperthermophilic archaeon Methanococcus jannaschii has been cloned into Escherichia coli. Extremely thermoactive and thermostable amylolytic activity was confirmed in partially purified enzyme solution. This enzyme exhibited a temperature optimum of 100 degrees C and a pH optimum pH 5.0-8.0. Hydrolysis of large 1,6-alpha- and 1,4-alpha-linked polysaccharides yielded glucose polymers of 1-7 units. Incubation with amylose displayed the highest activity. The catalyst was activated and stabilized by Ca2+ and exhibited extreme thermostability at 100 degrees C with a half-life of 78 h. PMID- 11898336 TI - An origin of DNA replication from streptomycete phage phi U1. AB - A DNA fragment from phage phi U1 containing an origin of DNA replication was identified. This fragment, designated ori, was able to support the maintenance in Streptomyces lividans of a plasmid lacking a functional Gram-positive ori. The sequence of the minimal ori fragment was determined and analyzed. The minimal fragment conferring replication origin function contained a number of direct and inverted repeats. The absence of an open reading frame in this ori fragment indicates that host factors alone were sufficient to initiate replication at ori. PMID- 11898338 TI - Morphological details of microorganisms revealed by RCH-microscopy at high magnification--a ready-to-use adaptation of a light microscope. AB - RCH-microscopy (Relief Contrast after Hostounsky) is a new method of optical microscopy in transmitted light developed with Lambda Ltd., Prague. This method was used to study bacteria, fungi including yeasts and algae at high magnification. The equipment provides a three-dimensional image of high contrast and resolution. The results of these microscopic observations can be used for both morphological (taxonomical) and ecological studies of microorganisms. PMID- 11898337 TI - Growth-associated production of poly-3-hydroxybutyrate by Bacillus mycoides. AB - Bacillus mycoides strain RIJ B-017, a growth-associated poly-3-hydroxybutyrate (PHB) producer was grown on sucrose-containing media. PHB accumulated in cells up to 72% of dry cell mass. The overall maximum value of PHB yield (Yp/s) and productivities (Qp and qp) 250 mgp/gs, 120 mgp L-1 h-1 and 30 mgp gx-1 h-1, respectively, were obtained at 15 g/L sucrose. Differential scanning calorimeter heating curve showed two peaks, one at 95.9 degrees C and another at 165.4 degrees C with a shoulder around 154.6 degrees C. The viscosity-average molar mass in chloroform at 27 degrees C was 505 kDa. The carbon content of PHB was 55.4% of the mass. PMID- 11898339 TI - Influence on Enterobacter cloacae metabolism, cell-surface hydrophobicity and motility of suprainhibitory concentrations of carbapenems. AB - The impact of postantibiotic effect (PAE) of carbapenems (imipenem, meropenem) on the metabolism (biosynthesis of macromolecules, respiration), cell-surface hydrophobicity and motility of a clinical isolate of Enterobacter cloacae was examined. The metabolism was evaluated after 16 h and after 1 d of cultivation using 2x and 4x minimum inhibitory concentrations (MIC) of both antibiotics for the induction of PAE. Imipenem at 4 x MIC did not induce PAE. After a 16-h cultivation (in the postantibiotic phase of both carbapenems), inhibition of nucleosynthesis and protein synthesis was found; after a 1-d cultivation, during regrowth stimulation of mainly 14C-leucine incorporation was found. The presence of the exogenous intermediates of citrate cycle, viz. 2-oxoglutarate, increased the respiratory activity of the cells. The cell-surface hydrophobicity (evaluated by three methods--bacterial adhesion to hydrocarbon, nitrocellulose-filter test and salt-aggregation test) decreased after PAE of both carbapenems; meropenem was more effective. Motility (an important virulence factor) was inhibited in the postantibiotic phase of both carbapenems; the 4 x MIC caused a higher inhibition. PMID- 11898340 TI - Phenolic acids reduce the genotoxicity of acridine orange and ofloxacin in Salmonella typhimurium. AB - Naturally occurring plant phenolics, p-coumaric acid (PA), caffeic acid (CA), ferulic acid (FA) and gentisic acid (GA) (25-100 nmol/L) had protective effects on acridine orange (AO; 216 mumol/L)- and ofloxacin (3 mumol/L)-induced genotoxicity in Salmonella typhimurium. FA, GA and CA exhibited a significant concentration-dependent protective effect against the genotoxicity of AO and ofloxacin, with the exception of PA, which at all concentrations tested abolished the AO and ofloxacin genotoxicity. UV spectrophotometric measurements showed the interaction of PA, FA, GA and CA with AO but not with ofloxacin; this interaction is obviously responsible for the reduction of AO-induced S. typhimurium mutagenicity. In the case of ofloxacin the antimutagenic effect of PA, FA, GA and CA is assumed to be a result of their ability to scavenge reactive oxygen species (ROS) produced by ofloxacin. PMID- 11898341 TI - Production, purification and characterization of intracellular alanylaminopeptidase of Pseudomonas sp. AB - The soil bacterium Pseudomonas sp. was found to synthesize an aminopeptidase that prefers Ala-beta-naphtylamide as substrate. The enzyme was purified 660-fold by ammonium sulfate fractionation, preparative electrophoresis, ion exchange chromatography on Protein-Pak Q 8 HR and molecular sieving chromatography on Zorbax SE-250. When purified to homogeneity, the enzyme was shown to be a monomeric protein with a molar mass of 65 kDa; it showed a maximum activity at pH 7.5 and 45 degrees C. PMID- 11898342 TI - Isolation and characterization of two types of actinophage infecting Streptomyces scabies. AB - Two types of actinophages, phi S and phi L, were isolated from soil samples by using Streptomyces scabies, a potato scab pathogen, as indicator strain. The phages were partially characterized according to their physicochemical properties, plaques and particles morphology, and their host range; this varied from narrow (for phi S) to wide (for phi L). The adsorption rate constants of the phi S and phi L were 3.44 and 3.18 pL/min, and their burst sizes were 1.61 and 3.75 virions per mL, respectively. One-step growth indicated that phi S and phi L have a latent period of 1/2 h followed by a rise period of 1/2 h. The temperate character of these phages was tested in other isolates of Streptomyces. Four of the phages (phi SS3, phi SS12, phi SS13 and phi SS17) were identified as temperate phages, since they were able to lysogenize SS3, SS12, SS13 and SS17. phi SS3, phi SS12 and phi SS13 were homoimmune, and they were heteroimmune with respect to phi SS17. The restriction barriers of lysogenic isolates (SS12, SS13 and SS17) interfered with the blockage of plaque formation by phages (phi SS12, phi SS13 or phi SS17) propagated on them, about 75% of lysogenic isolates had restriction systems. The exposure of the lysogenic isolates (SS12, SS13 and SS17) to UV-irradiation prevented the possible restriction barriers of these isolates so that these barriers could be overcome. PMID- 11898343 TI - Identification of DNA-binding proteins involved in regulation of expression of the Streptomyces aureofaciens whiH gene encoding a sporulation transcription factor. AB - Using the gel mobility-shift assay with protein fractions from different developmental stages of solid-grown Streptomyces aureofaciens, we identified two different proteins specifically bound to the whiH promoter region. Only one protein (RwhA) was detected in young substrate mycelium cultivated in liquid medium. On comparing the mobility of the resulting complexes, one of the bound proteins present in substrate mycelium and in early stages of aerial mycelium seemed to be identical with the RwhA. The other detected protein with a higher mobility (RwhB) was present in all developmental stages except for mature spores. DNA footprinting analysis localized the binding site of RwhB to nucleotides -23 to +40 relative to the transcription start point of the PwhiH promoter. RwhA from young substrate mycelium protected the DNA fragment from -106 to -77 in coding strand and -126 to -82 in noncoding strand. WhiH has homology to a large family of metabolism-related repressors and seems to regulate negatively its own expression. These observations (and the results of transcription analysis of the whiH gene obtained earlier) suggest that two different proteins influence the expression of whiH gene in S. aureofaciens. The putative repressor-like RwhA protein protects expression of whiH in substrate mycelium either in liquid medium or during differentiation. The other detected protein, RwhB, which binds to the whiH promoter region during differentiation, may represent two forms of WhiH, one with a repressor role at the beginning of differentiation and second with the role of activator at the time of sporulation. PMID- 11898344 TI - High-temperature hydrocarbon biodegradation activities in Kuwaiti desert soil samples. AB - Soil samples taken monthly from the Burgan South oil field of Kuwait for one year degraded crude oil, phenanthrene, and hexadecane. Bacteria were better degraders at high-temperature (55 degrees C) than fungi, especially in the drier, hotter months. Depending on the period of sampling, bacteria degraded hydrocarbons in the range of 46-86% (crude oil), 42-100% (hexadecane) and 5-58% (phenanthrene). Fungi alone accounted for degradation by 20-81% (crude oil), 30-95% (hexadecane) and less than 55% (phenanthrene). PMID- 11898345 TI - Organic fertilization changes the response of mycelium of arbuscular mycorrhizal fungi and their sporulation to mineral NPK supply. AB - The synergetic effect of organic (cow manure) and mineral fertilization on the development arbuscular mycorrhizal (AM) fungi was demonstrated. The length of AM mycelium and sporulation were used as sensitive markers of the physiological state of soil AM fungal population. In manured treatments, both parameters increased in proportion with increasing mineral fertilization. In unmanured soil, the opposite trend was observed for the length of AM hyphae, which decreased with increasing mineral fertilization. Correlation analysis showed the dependence of length of AM hyphae and sporulation on soil available phosphorus. The correlation was negative in soil with no mineral fertilization and positive in soil supplied with luxury doses of mineral fertilizer. PMID- 11898346 TI - Differences in the microflora of scarified and unscarified seeds of Karwinskia humboldtiana (Rhamnaceae). AB - Seeds of Karwinskia humboldtiana obtained from a 1997 collection in the locality of Villa de Gracia Nuevo (Leon, Mexico) were contaminated with spores of filamentous fungi, bacteria and yeasts. The concentration of microorganisms in unscarified seeds ranged from 3.0 x 10(3) to 7.5 x 10(3) CFU/g. Predominant were bacterial isolates of the genera Aeromonas sp., Bacillus, and Pseudomonas; from filamentous fungi were identified Alternaria, Aspergillus niger, Cladosporium sp., Fusarium sp., Mucor sp., Penicillium commune, Trichothecium sp.; from yeasts Rhodotorula sp. and Saccharomyces cerevisiae. Seed scarification significantly reduced the microbial contamination. Of the original fungal isolates, only two were identified on scarified seeds, viz. Cladosporium sp. and Saccharomyces cerevisiae; although a relatively high incidence of a unidentifiable of Penicillium sp. was found, the bacterial spectrum was not altered. Treatment of scarified seeds with Vitavax 200 WP and Pomarsol Forte 80 WP (3 mg/g seeds) augmented germination by 10-19% compared to treated unscarified seeds, and by 16 31% compared to untreated unscarified seeds. PMID- 11898348 TI - Coprophilous streptomycetes and fungi--food sources for enchytraeid worms (Enchytraeidae). AB - Food selection experiments demonstrated that Enchytraeus crypticus (Oligochaeta, Enchytraeidae) was attracted by Streptomyces species and microscopic fungi in vermiculture substrates and in the gut content of Eisenia andrei earthworms. Consumption of spores and/or mycelia of attractive strains influenced markedly the proliferation of E. crypticus. There was a 74-fold increase in the numbers of enchytraeids fed on the mixture of Aspergillus flavus and Verticillium tenerum mycelia or on mycelium of one strain of Streptomyces in reproduction tests. Lower rates of increase of E. crypticus (50-fold or less) were observed in variants where V. tenerum or mixtures of fungi and streptomycetes were offered as food. We showed a potential importance of microbial populations in vermicultures and indicated that their regulation may provide a way to increase the productivity of such systems. PMID- 11898347 TI - Effects of pH on the growth rate, motility and photosynthesis in Euglena gracilis. AB - The influence of pH 3-10 on the growth, motility and photosynthesis in Euglena gracilis was demonstrated during a 7-d cultivation. The cells did not survive at pH < 4 and > 8, highest growth rate being detected at pH 7. Motility followed a similar pattern as growth rate. Photosynthetic response curves were shown to be of the same type over the whole pH range. High respiration was characteristic for cells grown at pH 5 and 6, the lowest one at 7. At high and also at low pH more active respiration was found which can be considered as a protective response on proton stress. Respiration was not completely inhibited with potassium cyanide. Photosynthesis was the most effective at pH 6; lower and higher pH decreased photosynthetic efficiency. pH affected more the growth rate than the photosynthesis. PMID- 11898349 TI - Particulate 1,3-beta-D-glucan, carboxymethylglucan and sulfoethylglucan- influence of their oral or intraperitoneal administration on immunological respondence of mice. AB - The effect of orally or intraperitoneally administered particulate 1,3-beta-D glucan (PBG), carboxymethylglucan (CMG) or sulfoethylglucan (SEG), obtained from the culture filtrate of Saccharomyces cerevisiae, on the functions of murine peritoneal adherent cells (PC) (peroxidase activity, nitric oxide synthesis), on relative organ mass and on proliferation of splenocytes was determined. The modulating activities after parenteral and non-parenteral administration of these polysaccharides were compared. Significant enhancement of NO production was observed only after in vitro cultivation of PC in the presence of lipopolysaccharide (LPS) in groups of mice treated repeatedly orally with CMG, PBG and SEG at a dose of 50 mg/kg body mass. Peroxidase activity increased significantly after repeated oral administration of CMG and PBG at doses 150 and 50 mg/kg, SEG 150 mg/kg body mass. The peroxidase activity and NO synthesis in mice given a single intraperitoneal injection of glucans (15 mg/kg body mass) were slightly higher than those after oral administration. Neither a significant enhancement of relative organ mass nor enhancement of the proliferative response of splenocytes to in vitro added stimuli (LPS, phytohemagglutinin) after repeated oral or single intraperitoneal administration of beta-glucans was observed. PMID- 11898350 TI - The role of microflora in the development of intestinal inflammation: acute and chronic colitis induced by dextran sulfate in germ-free and conventionally reared immunocompetent and immunodeficient mice. AB - One-week dextran sulfate treatment of conventional (CV) immunodeficient (SCID) mice gave rise to acute colitis in the colon mucosa; germ-free (GF) SCID mice did not exhibit any changes in colon morphology. Dextran sulfate application to CV immunocompetent (BALB/c) mice did induce substantial changes in the colon mucosa (grade 4); GF BALB/c mice showed mild changes in the colon morphology (grade 1) only. GF SCID mice and CV SCID mice died during the second round of dextran sulfate treatment suffering from chronic colitis; GF BALB/c mice exhibited mild crypt distortion while CV BALB/c mice showed a complete loss of the surface epithelium (grade 4), accompanied by T and B lymphocyte infiltration. PMID- 11898351 TI - Bifidobacterium bifidum monoassociation of gnotobiotic mice: effect on enterocyte brush-border enzymes. AB - The effect of intestinal colonization with Bifidobacterium bifidum (Gram-positive anaerobic bacterium colonizing the intestine of healthy new-born mammals, exhibiting a probiotic effect, protecting the intestinal mucosa against colonization by pathogenic microflora) on enterocyte brush-border enzymes was examined in weaned 23-d- and in 2-month-old gnotobiotic inbred mice and compared with that in corresponding germ-free (GF) and conventional (CV) controls. The two groups of GF mice were associated with human B. bifidum 11 d before the end of the experiment. Specific activity of enterocyte brush-border enzymes--lactase, alkaline phosphatase and gamma-glutamyltranspeptidase was significantly higher in both age groups of GF mice in comparison with CV ones; on the other hand, sucrase and glucoamylase activities were higher in CV mice. Monoassociation with B. bifidum accelerates biochemical maturation of enterocytes resulting in a shift of specific activities of brush-border enzymes between the values found for GF and CV mice. This effect of B. bifidum supplementation was less pronounced for alkaline phosphatase, sucrase, glucoamylase and dipeptidyl peptidase i.v. in immature gut of weaned mice than of 2-month-old ones. PMID- 11898352 TI - [Current aspects of surgical treatment in anophthalmia]. AB - The purpose of this work was to determine the main trends in surgical rehabilitation of patients with anophthalmia and define the principles of treating patients with poor ocular prostheses. A total of 3345 interventions on the orbital cavity and eye appendages were carried out at plastic surgery and ocular prosthesis department in 1991-2000. Of these, 1535 patients (45.8%) were with anophthalmia. Analysis of disease structure in anophthalmia helped determine the priority trends of surgical rehabilitation of this group of patients: enucleation with plasty of the supporting motor stump, 36.6%; postradiation atrophy of orbital tissues after enucleation for retinoblastoma, 22.6%; anophthalmic syndrome, 11.7%; cicatricial deformation of the cavity, 11.5%; anophthalmia with concomitant eyelid deformation, 6.1%; anophthalmia with orbital deformation, 3.1%; denudation of orbital implants, 2.6%; congenital anophthalmia, 2.5%; prolapse of the inferior vault and upper eyelid ptosis, 1.8 and 1.4%, respectively. Individual methods of treatment and main rehabilitation principles were developed for each group of patients. PMID- 11898353 TI - [New modification of autoconjunctival plastic surgery in urgent corneal surgery]. AB - A new modification of plastic repair of deep defects of the cornea was performed: autoconjunctival plasty of the cornea with a flap on a pedicle with its rigid fixation in the corneal bed. The operation was carried out in 30 patients with infectious, trophic, and autoimmune involvement of the cornea, 24 of these with perforation of the cornea. The patients were observed for 1 year after the operation. The conjunctival-corneal complex formed in 24 cases, dislocation of the flap without its necrosis occurred in 2 cases, and necrosis of the flap in 4 cases. This modification of conjunctival plasty of the cornea can serve as the operation of choice in urgent ophthalmic surgery in cases when transplantation material is unavailable. PMID- 11898354 TI - [Laser extraction of brown cataracts with an Nd-YAG 1.44 mcm laser]. AB - A total of 172 extractions of brown nucleus cataracts with maximum compactness were carried out using a RAKOT complex of devices, developed by the authors for laser extraction of compact nuclei and based on the use of Nd:YAG laser with a wavelength of 1.44 mu. Brown cataract was effectively destroyed at 200-250 mJ and pulse generation frequency 25 Hz within 9-12 min during the first 20 operations and 4-6 min during subsequent 152 operations. Five (2.9%) ruptures of the posterior capsule of the lens were detected. The cornea remained transparent on days 1-2 in 9 of 20 (45%) first operations and in 150 of 152 (98.6%) subsequent operations. A combination of common and heavy viscous elastic was used for the protection of the posterior surface of the cornea. The posterior capsule of the lens was protected by an original silicone thin plate. Loss of corneal epithelium in such a method of nucleus destruction is no more than 2-6%. Intraocular pressure is no higher than 21 mm Hg. If the coefficient of liquid discharge easiness is decreased before the intervention, intraocular pressure can increase during the early postoperative period. High visual acuity (0.7-1.0) was attained in 98% cases in the absence of concomitant retinal diseases. No complications which could be attributed to negative effect of laser on the eye were recorded during the remote period after the operation. The results indicate high efficiency and safety of laser extraction of cataracts. PMID- 11898355 TI - [Possible reasons for hypercorrection and incomplete correction of refractive anomalies using photorefractive surgery]. AB - Probable variants of relationships between the major parameters of the eyeball, such as thickness of the cornea, its curvature radius, limbus diameter, and length of the anteroposterior axis of the eye are analyzed on a model representing the structure of external capsule of the eye with the use of schemes. Numerous combinations of these parameters of anatomical structures of the eye are responsible for hypercorrection and poor correction of refraction by examer laser surgery, by photorefraction keratectomy in particular. Along with well-known factors causing regression in remote periods after photorefraction keratectomy (corneal regeneration, effect of ophthalmic tone, accommodation spasm, etc.), individual variants of eye anatomy, neglected during laser exposure planning, can lead to unsatisfactory results of laser treatment. PMID- 11898356 TI - [Molecular diagnosis of retinoblastoma. First experience in Russia]. AB - The first experience with molecular diagnosis of retinoblastoma (RB) in Russia is presented. A protocol based on the use of up-to-date molecular diagnostic methods helps detect structural and functional abnormalities in RB1 gene, diagnose RB in disputable cases and at early stages of the disease. Forty-five families with various forms of RB were examined. Twenty-three mutations in various sites of RB1 gene were characterized. Abnormal methylation in the promotor area of RB1 gene was detected in 20% cases and loss of heterozysity by intragene microsatellite markers was detected in 70% cases. Hence, the causes of RB were detected in 80% families, which led to early diagnosis in close relatives of patients and helped evaluate the repeated risk of the tumor in families of patients with RB. PMID- 11898357 TI - [Effect of pilocarpine and cycloplegic substances on ophthalmic tone in healthy eyes and ones with glaucoma]. AB - The study was carried out on the eyes of patients (aged 20-75 eyes) without glaucoma and with primary open-angle glaucoma (POAG). A single instillation of 1 or 2% pilocarpin hydrochloride did not lead to a decrease of intraocular pressure (IOP) in the eyes without glaucoma. IOP fluctuations after pilocarpin instillations were accidental and as a rule were observed both in the treated and control eyes. Instillations of the same pilocarpin solutions significantly decreased IOP in the eyes with OAG in the majority of cases (in 76% in our study), which is apparently due to removal or decrease of the canalicular block. The results of relieving tonometric pilocarpin test should be taken account of in early diagnosis of glaucoma, but the diagnostic parameters of the test should be determined more precisely. After instillation of short-acting cycloplegic drugs (1% homatropin and 0.5% tropicamide) IOP did not change either in normal or POAG eyes. These drugs are recommended for dilatation of the pupil for ophthalmoscopy in patients with anterior chamber open angle glaucoma. PMID- 11898358 TI - [Prospects for developing treatment of uveal melanoma from the position of modern carcinogenesis concepts]. AB - Immunohistochemical studies of regulator proteins p53, Bcl-2, Bax, Ki-67, and CD 95L showed that apoptosis suppression, allowing unlimited proliferation of tumor cells, plays an important role in the carcinogenesis of uveal melanoma. This fact dictates the need of including drugs arresting suppression or inducing apoptosis in therapeutic protocols for uveal melanoma. Prognostic significance of p53, Bcl 2, Bax, and CD-95L was demonstrated. Each of the peptides can be used as a prognostic marker. PMID- 11898359 TI - [Electrophysiological methods for examining preterm children and diagnosis of retinopathy of prematurity]. AB - The first stage of the study was devoted to registration of visual evoked potentials (VEP) in 14 full-term and 27 preterm children without signs of neonatal retinopathy (NR) at the age of 6-7 months. The authors conclude that VEP in preterm children without signs of NR, born at different terms of gestation, reach the values of full-term children by the age of 6 months. These data should be taken account of when examining preterm babies for timely diagnosis of pathological changes in the visual analyzer and treatment. The second stage of the study was developed to registration of VEP and electroretinogram (ERG) in 36 children aged 4-12 years with cicatricial forms of NR. Changes in the central compartments of the retina were responsible for low visual functions in children with the cicatricial stages of NR. This abnormality is caused by displacement of the macular zone, impairment of its electrogenesis and topographic anatomic relationships. Examination of the marginal retinal periphery (superior external quadrant) once a year is obligatory for patients with cicatricial stages of NR in order to timely detect inapparent local fixed detachments or prevent them by timely prophylactic laser coagulation of the retina. PMID- 11898360 TI - [Clinico-etiologic features of various types of courses of suppurative corneal ulcer]. AB - Clinical observations of 366 patients with purulent corneal ulcers over the period of 1989-1998 helped the author distinguish two major patterns of its clinical course: acute (group 1, 164 patients) and primary chronic (group 2, 202 patients). The acute type was characterized by a fulminant onset, rapid progress of the ulcer, high incidence of complications (23.8%); growth of bacteria from ulcer surface was mainly due to highly pathogenic microflora (65.9%). Primary chronic type was characterized by a subacute onset, torpid course, lower incidence of complications (7.4%); the main etiological cause was opportunistic microflora (74.7%). PMID- 11898361 TI - [Clinico-morphological features of posterior post-traumatic uveitis]. AB - Time course of the clinical picture and echographic symptoms of posterior posttraumatic uveitis are described. Two patterns of posterior posttraumatic uveitis are presented: chronic with a favorable course, when inflammation at the posterior pole is arrested within 3-6 months, and chronic progressive with an unfavorable course, with rapidly developing occular subatrophy, progressive hypotone, loss of visual functions, and high risk of sympathic ophthalmia. Morphological changes in the eye membranes in chronic progressive posterior posttraumatic uveitis are described. PMID- 11898362 TI - [Clinical evaluation of the effectiveness of combined use of local and general hypotensive agents in patients with glaucoma and hypertension]. AB - Visual functions, hydrodynamics, and bloodflow in internal carotid arteries were evaluated in 60 patients with primary glaucoma and essential hypertension. Effects of systemic and local beta-adrenoblockers and combinations of local hypotensive agents with calcium antagonists and angiotensin-converting enzyme inhibitors were studied. Negative effect of a combination of local and oral beta adrenoblockers and positive effect of a combination of local hypotensive agents with oral capotene were revealed. Capotene is recommended for the treatment of glaucomatous patients with essential hypertension. PMID- 11898363 TI - [Results of treating thrombosis of retinal vessels in ambulatory care]. AB - The results of therapy of retinal vascular thrombosis are analyzed. Common therapy (tissue therapy, vasodilatants, angioprotectors, group B vitamins) was supplemented after discharge from hospital (for patients with thrombosis of the central retinal vein) by laser coagulation of the retina, 10 retrobulbar injections of heparin (0.1 ml) with dexasone (0.5 ml), then 10 intravenous injections of trental (0.5 ml), 10 intravenous injections of 2.4% euphylline, and intramuscular injections of lasix (4 ml, 3 injections every other day). Immediately after discharge the patients were prescribed ethanol solution of bee glue (40 drops orally in boiled water) and 1 tea-spoon of pollen with water, both to be taken after overnight fasting. Due to intensive care, hemorrhages in the retina resolved sooner and visual acuity increased to 0.7-0.9 diopters with correction; intraocular pressure 3.4 months after the disease onset was 18.21 mm Hg. One year after the disease, patients who received no retrobulbar injections of trental, been glue, or pollen still had hemorrhages along the vessels on the fundus oculi, retinal degeneration and new vessels in the paramacular area and on the optic disk; visual acuity of these patients was 0.02 diopters without correction. PMID- 11898365 TI - [A case of sarcoid uveitis as the earliest symptom of systemic sarcoidosis]. PMID- 11898364 TI - [Prevalence of refractive anomalies among school children]. AB - Complex ophthalmological examination of schoolchildren was carried out in 7 schools and 1 gymnasium. A total of 6027 children aged 6-17 years were examined (3188 boys and 2839 girls). Ocular disorders were detected in 1011 (16.8%), of these refraction abnormalities in 593 (58.7%) and accommodation spasm in 242 (23.9%). Myopic refraction was detected in 307 children (5.1%), medium hypermetropia in 150 (2.5%), high hypermetropia in 7 (0.1%), and astigmatism in 129 (2.1%). The main causes of visual disorders in schoolchildren are refraction abnormalities, primarily myopia. PMID- 11898366 TI - [A case of Dirofilaria repens helminthiasis under the conjunctiva]. PMID- 11898367 TI - [Ocular symptoms in multiple sclerosis]. PMID- 11898368 TI - [A. N. Maklakov--founder of Moscow Ophthalmological School]. PMID- 11898369 TI - [Clinical picture of open-angle glaucoma in patients with critical stenosis of the internal carotid artery]. AB - Clinical picture of primary open-angle glaucoma (POAG) was studied in 32 patients (57 eyes) with pronounced stenosis of the extracranial segment of the internal carotid artery (ICA) on the side of the glaucomatous eye. Major clinical signs were detected: conjunctival angiopathy, pronounced diffuse dystrophy of the iris, absence of pigmented iridal rim, pseudoexfoliations, pale optic disk with saucer like excavation of different severity. Intraocular pressure (IOP) was 26-34 mm Hg and could not be normalized by drug therapy or antiglaucomatous surgery. Ultrasonic methods for examination of the eyeball vessels showed pronounced decrease of the bloodflow velocity and a significant increase of resistance index in the orbital and central retinal arteries. Reconstructive operations on the carotid arteries performed in POAG patients for critical stenosis of ICA improved the eyeball hemoperfusion and decreased IOP. PMID- 11898370 TI - [Prospects for using the Mullerectomy for correcting ptosis of the upper eyelid]. AB - New anatomic topographic and clinical validation of mullerectomy is presented and more comprehensive evaluation of preoperative adrenergic drug test is suggested. Morphological substantiation of an isolated resection of the superior tarsal muscle (Muller muscle) and orbital conjunctiva of the upper eyelid with subsequent fixation of the proximal part of Muller muscle to the tarsal plate is presented. The results of surgical treatment of 51 patients evidence high reliability, functional and cosmetic efficiency of mullerectomy which does not involve a skin incision, and a low incidence of complications (mean period of observation 35 months). PMID- 11898371 TI - [Nebulizer therapy from the physical therapist's point of view]. AB - The author describes devices for drug inhalation (a dose-metered inhaler, dry powder inhaler, compressor pneumatic and ultrasonic nebulizers), their advantages and defects, indications and contraindications in patients with bronchial asthma and chronic obstructive pulmonary disease. PMID- 11898373 TI - [Status of microcirculation in elderly hypertensive patients during treatment with sodium chloride baths]. AB - Microcirculation was studied with biomicroscopy of the eye bulbar conjunctiva's vessels in 50 patients with essential hypertension stage II aged 60-80 years. They took half baths with sodium chloride mineral water at Irkutsk health resort "Angara". The treatment resulted in improvement of perivascular and intravascular end blood flow. PMID- 11898372 TI - [Effect of sinusoidal modulated currents on physical ability to work and disruption of cardiac rhythm in coronary heart disease with stable angina pectoris]. AB - 137 patients with coronary heart disease (CHD) and stable angina pectoris (functional classes I and II) participated in the study. 67 patients received a course of sinusoidal modulated currents (SMC) on the heart area; 40 patients--SMC on the sinocarotid zones; 30 patients received placebo procedures. The treatment outcomes were assessed with spiroveloergometry, ambulatory Holter ECG monitoring. The results demonstrate that course SMC provide a training effect (improved physical performance, coronary heart reserve) and antiarrhythmic action (mean daily number of ventricular extrasystoles decreased by 36.6-50.4%, supraventricular extrasystoles by 42.7-43.2%). PMID- 11898374 TI - [Dynamics of vegetative indicators induced by low-frequency magnetotherapy and EHF-puncture in hypertensive workers exposed to vibration]. AB - Low-frequency magnetic fields and EHF-therapy have been used in correction of autonomic homeostasis in workers exposed to vibration for different periods of time. The workers suffered from early arterial hypertension. Vegetative status and central hemodynamics improved best in workers exposed to vibration for less than 5 years. If the exposure was 6-15 years, a positive trend occurred in the tension of regulatory mechanisms. Workers with long exposure to vibration suffering from vagotonia showed an inadequate response of the autonomic parameters to treatment. This necessitates enhancement of therapeutic measures with medicines. PMID- 11898375 TI - [Antiatherogenic properies of various types of carbonate mineral drinking waters]. AB - Carbonate mineral waters of different types (Shmakovskaya, Sinegorskaya, Utserskaya) were examined in experimental model of hyperlipidemia for action on atherogenically altered lipid spectrum of blood and oxidative resistance of plasma. The above waters differ in mineralization and specific components. Carbonate waters were found to have antiatherogenic properties and positive action on lipid indices and antiradical defense. Specific features of different carbonate waters enable a differentiated approach to correction of different variants of lipid disorders. PMID- 11898376 TI - [Effect of combined use of sulfate-sodium chloride mineral water and captopril in experimental atherosclerosis]. AB - Course effects of mineral water (MW) Dzhemukhskaya and captopril were studied in 80 Wistar male rats with experimental atherosclerosis. Captopril significantly enhanced effects of MW. Aldosterone levels decreased 2.2 times, triiodothyronine- by 57.2%, atherogenic index--2 fold. ACTH and hydrocortisone concentrations noticeably went up. High glucose levels in the blood diminished. Elevated levels of AlT and AsT normalized. Favourable shifts in combined use of MW and captopril in various doses were nearly the same, in some cases lower dose of captopril was more effective. PMID- 11898377 TI - [Color-physiopuncture corrections of autonomic disorders]. AB - Investigations have shown that electromagnetic waves of color band used for physiopuncture correction of functional-dynamic condition of channel-meridional system of the body in young patients with vegetovascular asthenia of a hypertensive type lead to a regress of the clinical symptoms, normalization of arterial pressure, correction of cerebral hemodynamic disorders, recovery of normal bioelectric activity of the brain. PMID- 11898378 TI - [Use of lidase photophoresis in lumbrosacral scar-adhesive process in postoperative rehabilitation of patients with spondylogenic neuropathy]. PMID- 11898379 TI - [Water--a key molecule in the effect of therapeutic physical factors]. AB - The article presents current conceptions of the structure, physicochemical properties and biological functions of water in terms of their physiotherapeutic value. Water may be essential not only in mechanisms of action of therapeutic physical factors but also in their selective absorption by tissues. Information is given on changes in water properties in response to its exposure to electric currents, ultrasound, magnetic and electromagnetic fields, laser radiation and other physiotherapeutic factors. PMID- 11898380 TI - [Interference therapy in combined treatment of children with childhood cerebral palsy]. PMID- 11898381 TI - [Combined ultrasound and sinusoidal modulated currents in treating children with juvenile rheumatoid arthritis]. AB - The exposure of the joints to ultrasound and sinusoidal modulated currents in combination with radon baths proved highly effective in slowly progressive juvenile rheumatoid arthritis of the first degree of activity. PMID- 11898382 TI - [Health of school children and their constitutional development in different organization of learning in school]. PMID- 11898383 TI - [Health improvement in children with risk factors for alcoholism]. PMID- 11898384 TI - [Hydrocarbonate-sodium chloride mineral water in rehabilitating children with atopic dermatitis]. PMID- 11898385 TI - [Experience in using diadynamic currents in patients with acute rhinitis]. PMID- 11898386 TI - [Therapeutic physical culture in complex sanitorium treatment of scoliosis in children and adolescents]. PMID- 11898389 TI - Embryonic stem cells provide a powerful and versatile model system. AB - Embryonic stem (ES) cells are pluripotent stem cells that differentiate both in vitro and in vivo into cell types derived from each of the three embryonic germ layers. ES cells and their close relatives, embryonal carcinoma (EC) cells and embryonic germ (EG) cells, have been used extensively as model systems for studying early mammalian development. This work has led to important insights into the mechanisms that control embryogenesis at the molecular and cellular levels. This chapter focuses on the use of ES cells as an in vitro model system for studying cellular differentiation and reviews several areas where important progress has been made. Impressive progress has been made in the isolation and characterization of ES cells from many species, including humans. Significant progress has also been made in the development of culture conditions that help direct the differentiation of ES cells to specific cell types that form during myogenesis, angiogenesis, hematopoiesis, neurogenesis, and cardiogenesis. The ability to inactivate virtually any gene in ES cells by gene targeting has vastly improved our understanding of the roles played by specific genes at the cellular and organismic levels. Moreover, ES cells and EC cells have been used widely to investigate how specific genes are turned on and turned off in the course of differentiation. In this connection, DNA array technology has been used to identify genes regulated when ES cells differentiate. The final section of this chapter discusses how work with ES cells is shaping our understanding of stem cells, mammalian development, and cell replacement therapy. PMID- 11898390 TI - Tissue culture models for studies of hormone and vitamin action in bone cells. AB - Osteoporosis is a major health care concern and levies a serious financial burden on the world health care system. For this reason, many physicians and scientists are engaged in research to better understand and treat this disease. To this end, numerous in vitro bone cell models have been developed to explore the cellular and molecular mechanisms of skeletal biology and for the identification and characterization of new drug targets and therapies. In this chapter, we review many of these cellular models as tools to study the hormonal regulation of bone metabolism. In particular, we pay special attention to new human bone cell models, since these have the greatest relevance to osteoporosis research and drug discovery. These new models include (1) the use of peripheral blood mononuclear cells as progenitors of osteoclasts and primary cultures of mesenchymal stem cells as precursors of osteoblasts; (2) the development of conditionally immortalized preosteoclastic and osteoblastic cell lines using temperature sensitive large T-antigens; and (3) the establishment of the first osteocytic cell lines. Thus, we now have at our disposal many good in vitro models to investigate the regulation of bone resorption and formation by hormones, vitamins and drugs. These models should accelerate our understanding of bone physiology and pathophysiology as well as our ability to develop important new therapies to prevent and treat skeletal diseases. PMID- 11898391 TI - Transport of leukotriene C4 and structurally related conjugates. AB - Transport proteins control the release of the endogenous glutathione conjugate leukotriene C4 (LTC4) from leukotriene-synthesizing cells as well as its hepatobiliary and renal elimination. The photolabile conjugated triene structure of LTC4 has enabled direct photoaffinity labeling of the multidrug resistance protein 1 (MRP1, symbol ABC C1) in membranes from mastocytoma cells, leading to the identification of the function of this protein as an ATP-dependent export pump for LTC4 and structurally related conjugates. MRP1 is assigned to the C branch of the superfamily of ATP-binding cassette (ABC) transporters and was originally identified by virtue of its association with drug resistance in tumor cells. Besides LTC4, which is a high-affinity substrate, a variety of conjugates of hydrophobic endogenous or xenobiotic substances with glutathione, glucuronate, or sulfate are transported by MRP1. In addition, hydrophobic compounds may undergo cotransport with glutathione. Effective inhibitors of MRP1-mediated transport include structural analogs of LTC4 and of other cysteinyl leukotrienes. The ATP-dependent transport system which transports cysteinyl leukotrienes across the hepatocyte canalicular membrane into bile was cloned and characterized as the second isoform or paralog of the mammalian MRP family, MRP2 (ABC C2). MRP2 is localized to the apical membrane of polarized cells. The overall substrate specificities of MRP1 and MRP2 are similar, despite an amino acid identity of only 48%. The transport proteins mediating the uptake of LTC4 into hepatocytes across the basolateral membrane are members of the organic anion transporter (OATP) branch of the solute carrier (SLC) superfamily and are thus distinct from the ATP-dependent export pumps of the MRP family. PMID- 11898392 TI - Interleukin-1 beta exerts a myriad of effects in the brain and in particular in the hippocampus: analysis of some of these actions. AB - The realization, in the past decade or so, that bidirectional communication between the central nervous system and the immune system was likely has sparked an explosion of interest in the roles certain cytokines, particularly the proinflammatory cytokine interleukin-1 beta (IL-1 beta), might play in the brain. The observation that IL-1 type I receptor was expressed in highest density in the hypothalamus was of significance in identifying a role for IL-1 beta in neuroendocrine modulation. However, the finding that receptor expression was also high in the hippocampus, an area of the brain which plays a pivotal role in memory and learning, has led to uncovering a role for IL-1 beta in cognitive function. There is now a great deal of evidence suggesting that IL-1 beta plays a significant role in hippocampal synaptic function, and the possibility that IL-1 beta may trigger some of the detrimental changes in certain neurodegenerative diseases is currently being assessed. The review addresses some of the issues relating to the role of IL-1 beta in the brain, specifically in the hippocampus. PMID- 11898393 TI - Leptin and sweet taste. AB - Leptin, the product of the obese (ob) gene, is a hormone primarily produced in adipose cells, and also at smaller amounts in some other peripheral organs. It regulates food intake, energy expenditure, and body weight. Leptin is thought to promote weight loss, at least in rodents, by suppressing appetite and stimulating metabolism. Mutant mice that lack either leptin or functional leptin receptors, such as ob/ob and db/db mice, are hyperphagic, massively obese, and diabetic. Central hypothalamic targets are mainly responsible for the effects of leptin on food intake and weight loss. However, there are also direct effects on peripheral tissues. Recently, the taste organ was found to be one of the peripheral targets for leptin. The hormone specifically inhibits sweet taste responses in lean mice and not in db/db mice. Thus leptin appears to act as a modulator of sweet taste, provided a functional leptin receptor is expressed by the taste cells. This chapter reviews the genetics and molecular biology of leptin and its receptors, the receptor mechanisms for sweet taste, the modulating action of leptin on taste receptor cells, and the consequences for the regulation of food intake. PMID- 11898394 TI - Molecular, structural, and cellular biology of follitropin and follitropin receptor. AB - Follitropin and the follitropin receptor are essential for normal gamete development in males and females. This review discusses the molecular genetics and structural and cellular biology of the follitropin/follitropin receptor system. Emphasis is placed on the human molecules when possible. The structure and regulation of the genes for the follitropin beta subunit and the follitropin receptor is discussed. Control of systemic and cellular protein levels is explained. The structural biology of each protein is described, including protein structure, motifs, and activity relationships. Finally, the follitropin/follitropin receptor signal transduction system is discussed. PMID- 11898395 TI - Factor VIIa/tissue factor-induced signaling: a link between clotting and disease. AB - Tissue factor is a cellular receptor for plasma clotting factor VII. In health, tissue factor is constitutively expressed in many cells, such as fibroblasts and keratinocytes, but is absent in vascular cells that come in contact with blood. However, tissue factor is induced in vascular cells in certain pathophysiological conditions. Thus, vessel wall injury or a disease condition allows blood to come in contact with tissue factor, resulting in factor VII binding to tissue factor. Once native factor VII complexed with tissue factor is converted to the enzyme factor VIIa, the complex triggers the clotting cascade that ultimately leads to fibrin formation. In addition to its established role in coagulation, molecular links between factor VIIa/tissue factor and various biological processes, such as development, inflammation, and tumor metastasis, are also evident. Recent studies suggest that factor VIIa/tissue factor affects various cellular processes by inducing intracellular signaling. Emerging evidence suggest that factor VIIa/tissue factor participates in cell signaling by two distinct mechanisms: proteolysis-independent signaling via the cytoplasmic domain of tissue factor, and proteolysis-dependent signaling, which is independent of tissue factor's cytoplasmic tail. In proteolysis-independent signaling, filamin 1 is recruited to tissue factor upon its ligation, and this probably provides an essential intracellular link in transmitting signals from tissue factor. In proteolysis dependent signaling, factor VIIa/tissue factor activates one or more protease activated receptors, which couple to G proteins, to impact multiple signaling pathways. In this chapter, we review various nonhemostatic functions attributed to factor VIIa and tissue factor, describe signaling mechanisms initiated upon factor VIIa binding to tissue factor, and discuss how factor VIIa/tissue factor induced signaling could contribute to various pathophysiological processes. The relationship between increased clotting and manifestation of various diseases is well recognized. Although aberrant activation of the coagulation pathway is primarily the consequence of a disease, activation of the coagulation pathway during the disease process, in turn, could contribute to pathogenesis of the disease. Further, recent transgenic studies in mouse suggest that the coagulation system also plays a role in embryogenesis and development. Then the question arises, how do proteins involved in the clotting regulate cellular functions? The coagulant proteases can regulate cell behavior in a number of ways. They can function like hormones, activating multiple signaling pathways, as well as release bioactive fragments and growth factors by proteolysis of cell-surface and extracellular matrix (ECM) components. This chapter reviews our current understanding of the role of factor VIIa/tissue factor (VIIa/TF), an enzyme/cofactor complex that triggers the clotting cascade, in affecting various cellular functions not related to the clotting, and discusses potential mechanisms by which it regulates cellular functions. PMID- 11898396 TI - Antiproliferative action of vitamin D. AB - During the past few years, it has become apparent that vitamin D may play an important role in malignant transformation. Epidemiological studies suggest that low vitamin D serum concentration increases especially the risk of hormone related cancers. Experimentally, vitamin D suppresses the proliferation of normal and malignant cells and induces differentiation and apoptosis. In the present review we discuss the mechanisms whereby vitamin D regulates cell proliferation and whether it could be used in prevention and treatment of hyperproliferative disorders like cancers. PMID- 11898397 TI - Molecules in blastocyst implantation: uterine and embryonic perspectives. AB - Synchronized development of the embryo to the active stage of the blastocyst, differentiation of the uterus to the receptive state, and a "cross talk" between the blastocyst and uterine luminal epithelium are essential to the process of implantation. In spite of considerable accumulation of information and the present state of the knowledge, our understanding of the definitive mechanisms that regulate these events remains elusive. Although there are species variations in the process of implantation, many basic similarities do exist among various species. This review focuses on specific aspects of the implantation process in mice with the hope that many of the findings will be relevant to the process in humans. To establish signaling mechanisms of embryo-uterine interactions during implantation, studies on both embryonic and uterine consequences are required to generate more meaningful information. Due to ethical restriction and experimental limitation, it is difficult to generate such information in humans. This review has attempted to provide a comprehensive, but not complete, narration of a number of embryonic and uterine factors that are involved in the process of implantation in autocrine, paracrine, and/or juxtacrine manners in mice at the physiological, cellular, molecular, and genetic levels. PMID- 11898398 TI - Microarray analysis of B-cell stimulation. AB - B-cell development to antibody-producing plasma cells requires the concerted function of a large number of genes and proteins. Genome-level expression profiling during human B-cell maturation was studied in anti-immunoglobulin M stimulated Ramos cells. cDNA microarrays were used to follow changes in the transcriptome over several days. Close to 1500 genes had significantly altered expression at least at one time point. The genes were organized into clusters based on expression profiles and were further characterized based on the functions of the coded proteins. Several groups of genes important for B cells were analyzed. Here we concentrate on genes involved in signal transduction and cytokines and their receptors. The results provide knowledge on the development of humoral immunity. Several new genes were found to be essential for B-cell development. They can be used as targets for research and possibly for drug development. PMID- 11898399 TI - Estimation of individual types of glutathione peroxidases. PMID- 11898400 TI - High-throughput 96-well microplate assays for determining specific activities of glutathione peroxidase and thioredoxin reductase. PMID- 11898401 TI - Selenoprotein P. PMID- 11898402 TI - Iodothyronine deiodinases. PMID- 11898403 TI - Expression and regulation of thioredoxin reductases and other selenoproteins in bone. AB - The expression of thioredoxin reductases and other selenoproteins in cells of the bone microenvironment may represent an important means of regulation of bone resorption and remodeling in health and disease. Selenoproteins and their substrates may influence intracellular and extracellular redox-dependent signaling, transcription factor activity, posttranslational modification of proteins, and general or compartmentalized scavenging from ROIs. However, the evaluation of their biological role in bone and their potential in terms of therapeutic approaches is just beginning. PMID- 11898404 TI - Selenocysteine insertion sequence element characterization and selenoprotein expression. PMID- 11898405 TI - Selenoprotein W. PMID- 11898406 TI - Genetic and functional analysis of mammalian Sep15 selenoprotein. PMID- 11898407 TI - Selenocysteine lyase from mouse liver. PMID- 11898408 TI - Selenocysteine methyltransferase. PMID- 11898409 TI - Phospholipid-hydroperoxide glutathione peroxidase in sperm. PMID- 11898410 TI - In vivo antioxidant role of glutathione peroxidase: evidence from knockout mice. PMID- 11898411 TI - Recombinant expression of mammalian selenocysteine-containing thioredoxin reductase and other selenoproteins in Escherichia coli. PMID- 11898412 TI - Mammalian thioredoxin reductases as hydroperoxide reductases. PMID- 11898413 TI - Transfer RNAs that insert selenocysteine. PMID- 11898415 TI - Trypanothione and tryparedoxin in ribonucleotide reduction. PMID- 11898414 TI - Tryparedoxin and tryparedoxin peroxidase. PMID- 11898416 TI - Selenium- and vitamin E-dependent gene expression in rats: analysis of differentially expressed mRNAs. PMID- 11898417 TI - Thioredoxin. Overview. PMID- 11898418 TI - Thioredoxin and glutaredoxin isoforms. PMID- 11898419 TI - Mammalian thioredoxin reductases. PMID- 11898420 TI - Selenoprotein biosynthesis: purification and assay of components involved in selenocysteine biosynthesis and insertion in Escherichia coli. PMID- 11898421 TI - Mitochondrial thioredoxin reductase and thiol status. PMID- 11898423 TI - Recycling of vitamin C by mammalian thioredoxin reductase. PMID- 11898422 TI - Protein electrophoretic mobility shift assay to monitor redox state of thioredoxin in cells. PMID- 11898424 TI - Thioredoxin cytokine action. PMID- 11898425 TI - Identification of thioredoxin-linked proteins by fluorescence labeling combined with isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 11898426 TI - Thioredoxin and mechanism of inflammatory response. PMID- 11898427 TI - Redox state of cytoplasmic thioredoxin. PMID- 11898428 TI - Thioredoxin, thioredoxin reductase, and thioredoxin peroxidase of malaria parasite Plasmodium falciparum. PMID- 11898429 TI - Human placenta thioredoxin reductase: preparation and inhibitor studies. PMID- 11898430 TI - Classification of plant thioredoxins by sequence similarity and intron position. PMID- 11898431 TI - Purification and analysis of selenocysteine insertion sequence-binding protein 2. PMID- 11898432 TI - Ferredoxin-dependent thioredoxin reductase: a unique iron-sulfur protein. PMID- 11898433 TI - Plant thioredoxin gene expression: control by light, circadian clock, and heavy metals. PMID- 11898434 TI - Thioredoxin genes in lens: regulation by oxidative stress. PMID- 11898435 TI - Thioredoxin overexpression in transgenic mice. PMID- 11898436 TI - Multiplex reverse transcription-polymerase chain reaction for determining transcriptional regulation of thioredoxin and glutaredoxin pathways. PMID- 11898438 TI - Nonsense-mediated decay: assaying for effects on selenoprotein mRNAs. AB - If the abundance of a particular selenoprotein mRNA is reduced during selenium deprivation, then the mRNA is likely to be a natural substrate for NMD. One assay for NMD involves changing the TGA Sec codon(s) to either a TGC cysteine codon or a TAA nonsense codon. If selenium deprivation elicits NMD and has no other effect on selenoprotein gene expression, then, regardless of selenium concentration, the level of UGC-containing mRNA should be most abundant, the level of UGA-containing mRNA should be intermediate in abundance, and the level of UAA-containing mRNA should be least abundant. Furthermore, the level of UGA-containing mRNA should be decreased by a decrease in selenium concentration, while the levels of UGC- and UAA-containing mRNAs should be unaffected by selenium concentration. A different assay for NMD involves coexpression of the particular selenoprotein gene and a vector expressing a dominant-negative version of hUpf1p. This assay is simpler and more versatile than the first assay because it can be used to assay any cellular gene in situ provided the cells can be stably transfected with the hUpf1p expression vector. PMID- 11898437 TI - Redox regulation of cell signaling by thioredoxin reductases. PMID- 11898439 TI - Novel selenoproteins identified from genomic sequence data. PMID- 11898440 TI - Semisynthesis of proteins containing selenocysteine. PMID- 11898442 TI - [Multiple organ failure and perioperative hemodynamic optimization in high risk surgical patients: is it an efficient strategy?]. PMID- 11898443 TI - [Hemodynamics of the cirrhotic patient during liver transplantation. Influence of the preservation of portal and vena cava flow]. AB - OBJECTIVE: To describe the hemodynamic pattern of patients undergoing liver transplantation with preservation of portocaval flow. PATIENTS AND METHODS: A prospective study of 20 cirrhotic patients who had not previously undergone surgery for portal hypertension or had porto-systemic bypass, both of which have hemodynamic effects in the cirrhotic patient. The patients were transplanted with preservation of inferior vena cava flow and temporary portocaval shunt. RESULTS: The decrease in cardiac output during the anhepatic phase was only 10% and mean blood pressure (77.6 +/- 11 versus 76 +/- 10 mm Hg) and supply pressures (central venous pressure 9.1 +/- 5.5 versus 8.4 +/- 5.3 mm Hg; pulmonary capillary pressure 11.4 +/- 6.1 versus 11.3 +/- 7.4 mm Hg) remained stable. Likewise, no significant increase in systemic vascular resistance (614 +/- 223 versus 676 +/- 306 dyne-sec/cm5) or heart rate (90 +/- 14 versus 97 +/- 17 beats/min). The number of units of packed red cells was 2.7 +/- 2.5 and 35% of the patients required no transfusions. Diuresis was stable throughout the procedure (total diuresis 3.6 +/- 2.4 mL/Kg/h; anhepatic phase 1.3 +/- 1.5 mL/Kg/h). CONCLUSIONS: Creation of a portocaval shunt during the anhepatic phase of liver transplantation allows hemodynamic vital signs to be held stable, decreases the need for transfusion and maintains diuresis. PMID- 11898441 TI - Mammalian selenoprotein gene signature: identification and functional analysis of selenoprotein genes using bioinformatics methods. PMID- 11898444 TI - [Prophylaxis of infective complications of central venous catheters]. AB - Infections related to central venous cannulation present first-magnitude problems in recovery rooms and intensive care units. Catheter-related bloodstream infection (CRBSI) is the most serious complication because of its high frequency and a mortality rate that averages around 3%. Although infections arise for various reasons, point-of-insertion contamination is the main cause when catheters are implanted for periods of less than 10 days. Contaminating microorganisms (especially Staphyloccocus epidermidis) find refuge from the host's defenses in a biofilm that covers the catheter. Several factors participate in the formation of this biofilm, such as catheter composition, proteins of the host or type of microorganism. Biofilm bacteria are resistant to both antibiotics and the host's own defenses (e.g. phagocytes and antibodies). The microorganism can then begin to reproduce, possibly leading to bloodstream infection. The measures designed to prevent this process include recommendations for both catheter insertion and maintenance. Recent meta-analyses have led to certain conclusions but no unanimity among authors. Thus, there is agreement on the adoption of strict aseptic technique during catheter insertion, on the use of chlorhexidine as a skin antiseptic and on choice of the subclavian vein. Such measures significantly decrease the frequency of CRBSI. Maintenance techniques that have been shown to be effective are the use of connectors impregnated with antiseptic, catheters impregnated with antiseptics or antibiotics, and permeable dressings. Additionally, building an experienced infusion-therapy team to insert and maintain central venous catheters has been shown to be one of the most effective measures for preventing CRBSI. PMID- 11898445 TI - [Late rupture of central catheter implanted through subclavian route]. AB - A 68-year-old woman with a subcutaneous reservoir and central venous catheter implanted in the left subclavian vein for chemotherapy developed a late fracture of the catheter 220 days after implantation, with migration of the distal fragment to the pulmonary artery. After diagnosis of the complication, the distal fragment had to be removed by radiologic intervention and the reservoir was withdrawn. The need for central venous treatments, whether for definitive parenteral nutrition or prolonged antibiotic infusion, pain therapy, chemotherapy or, lately, for hemodialysis has made the placement and use of implantable systems a routine procedure for anesthesiology services. However, such techniques are not free of multiple complications, some of which are serious and which may be caused by poor technique during insertion of the catheter. We consider careful selection of the point of subclavian vein insertion to be important. The scheduling of follow-up chest films is also crucial for ruling out a "pinch-off phenomenon". PMID- 11898446 TI - [Cervical epidural anesthesia with 0.75% ropivacaine in shoulder surgery]. AB - We present the cases of three patients scheduled for shoulder surgery under cervical epidural anesthesia with 0.75% ropivacaine. The technique was successful and surgery proceeded uneventfully in all three cases. The total doses of ropivacaine infused were 67, 90 and 109 mg. Premedication with intravenous atropine is recommended. Cervical epidural anesthesia offers several advantages over general anesthesia and other regional techniques for shoulder surgery: postoperative analgesia, lower total dose of the local anesthetic and single needle insertion with no need to elicit paresthesias or muscle movements. This technique brings about hemodynamic and respiratory changes in function of the extension and intensity of the block. Extent of the blockade to the upper thoracic sensory segments causes a total or partial sympathetic block with decreased heart rate, blood pressure and cardiac output. Limiting the initial and subsequent doses to restrict the sensory blockade to the surgical area will reduce hemodynamic complications. Ropivacaine provides an effective sensory block and a restricted motor block, reducing the probability of the restrictive pulmonary syndrome associated with cervical epidural anesthesia. PMID- 11898447 TI - [Lung transplantation in patients in mechanical ventilation before transplantation]. AB - Mechanical ventilation before lung transplantation has been identified as a risk factor for early death after surgery. However, several studies have reported patient series in which ventilation assistance was given preoperatively without increasing the rates of postoperative complications and death, apart from increasing time of postoperative intubation. The present retrospective analysis of the postoperative course of patients who had been mechanically ventilated before transplantation encompasses a period of 5 years in our hospital. Eight transplants (7 double- and 1 single-lung procedures) were performed. Six patients (75%) required extracorporeal oxygenation during surgery. Three patients (37.5%) died within 30 days of receiving the transplanted lung. The mean time of intubation after the operation was 10.3 days and the mean stay in the postoperative recovery and intensive care unit was 27.5 days. The most common postoperative complications were respiratory colonization (100%), with infection in 3 patients, and reimplantation injury (50%). Pretransplant mechanical ventilation was associated with high risk in the patient series we report; however, the survival rate observed suggest that such patients should be considered acceptable candidates to receive grafts if indicated. PMID- 11898448 TI - [Extrapulmonary tuberculosis in a patient with septic shock]. AB - A 61-year-old woman who was negative for type 1 human immunodeficiency virus developed vertebral osteomyelitis and skin lesions due to sepsis by Staphylococcus aureus. Microscopic examination of the skin showed alcohol resistant acid-fast bacilli. A polymerase chain reaction (PCR) assay for Mycobacterium tuberculosis was positive for skin and spinal samples, although the cultures were negative. The diagnosis of M. tuberculosis infection is difficult, particularly when the disease is extrapulmonary. Rapid diagnostic tests that use PCR identify the DNA of the bacillus with greater sensitivity than microscopic examination and can give results within 24 hours of receipt of a sample. We analyze the utility of PCR for diagnosing extrapulmonary tuberculosis. PMID- 11898449 TI - [Indications for electrocardiogram in the preoperative assessment for programmed surgery]. AB - OBJECTIVES: To establish indications for ordering a screening electrocardiogram (ECG) before scheduled surgery. To study the prevalence of abnormalities found in routine ECGs and the impact of routine ECGs on anesthetic and surgical management and on preventing perioperative complications. MATERIAL AND METHODS: A prospective study of 413 patients undergoing scheduled non-cardiac surgery over a two-year period. ECG anomalies were defined as major or minor in function of their association with perioperative morbimortality. ECG results were considered expected or unexpected in function of agreement with a patient's history. RESULTS: An ECG was done for all patients before surgery. Anomalies were observed in 41.9% of the ECGs, 28.6% of which were considered major. The prevalence of anomalies was greater among men over 40 years of age, with heart or respiratory disease and these classified as ASA III-V. The anomalies were unexpected in 8.9% and did not cause postponement or cancellation of scheduled procedures. Anomalies found led to changes in preoperative approach in 0.5% of the cases. Intraoperative complications were seen in 7.9% and postoperative complications in 24.6%. CONCLUSIONS: Preoperative ECGs should be obtained only in patients over 40 years of age who present cardiac or respiratory signs or symptoms and who are diagnosed of some heart or respiratory disease. PMID- 11898450 TI - [The use of laryngeal mask in abdominal surgery in a patient with myasthenia gravis]. PMID- 11898451 TI - [Platelet aggregation inhibitors and spinal anesthesia]. PMID- 11898452 TI - [Unilateral hearing loss after subarachnoid anesthesia]. PMID- 11898453 TI - [PAXPRESS pharyngeal airway: a new device for maintaining the airway during general anesthesia]. PMID- 11898454 TI - [The use of laryngeal mask in a female patient with sclerodermia]. PMID- 11898455 TI - [Usefulness of monitoring with BIS in myotonic dystrophy]. PMID- 11898456 TI - Eco-epidemiological characteristics of an unstable peri-urban focus of falciparum malaria. AB - Two fever surveys were carried out in Shahbad dairy, Delhi in the post-monsoon months of October and November, 1996. Shahbad dairy is a peri-urban locality in the northern periphery of Delhi. The prevalence of fever was found to be 11.6% in October and 2% in November. In the two surveys a total of 21 (28%) fever cases were found infected with malaria (17 with P. falciparum and 4 with P. vivax). The prevalence of malaria in the surveyed population was lower (25%) in October than in November (36.8%). More adults and males suffered from malaria than other age groups and the females. In November the mean asexual parasitaemia, for P. falciparum infection, sharply declined among 5-14-years old children by 68%, but among adults it increased by 32%. Collection of adult mosquitoes, from human houses, revealed the presence of 5 species of anopheline mosquitoes comprising Anopheles culicifacies, An. stephensi, An. annularis, An. subpictus, and An. nigerrimus, and a culicine mosquito Culex vishnui complex. Extensive breeding of these mosquitoes was detected in the vicinity of Shahbad dairy. Characteristic ecological support system (ESS) for malaria transmission was identified at the peri-urban focus of Shahbad dairy. PMID- 11898457 TI - Serodiagnosis of extra pulmonary tuberculosis using A-60 antigen. AB - The diagnosis of extrapulmonary tuberculosis including neurotuberculosis is difficult because of the low yield of culture positivity for Mycobacterium tuberculosis (M. tb). Serodiagnosis has emerged as a useful aid to the diagnosis of extrapulmonary tuberculosis. The utility and efficacy of detection of antimycobacterial antibodies to A-60 antigen in serum and/or cerebrospinal fluid (CSF) was analysed in 100 patients-neurotuberculosis-72, abdominal tuberculosis 12 and others-16. The overall positivity rate for the test was 75%. The positivity rate of the test in serum and/or CSF was 79.2% (57 of 72) in neurotuberculosis and 62.5% (10 of 16) for other forms of extrapulmonary tuberculosis. The positivity rate for antimycobacterial antibodies was higher for patients with tubercular meningitis -94.7%. To conclude, testing for antimycobacterial antibodies to A-60 antigen is a useful adjunct in the diagnosis of extrapulmonary tuberculosis especially neurotuberculosis. PMID- 11898458 TI - Host preference of Culex quinquefasciatus in Raipur city of Chattisgarh state. AB - Indoor resting freshly engorged Culex quinquefasciatus collected from three different habitats viz., human, cattle and mixed dwellings of six localities of Raipur city, were analyzed for the source of blood meal by precipitin test. Of the 60 specimens from human dwellings 52 were positive for human blood with an anthropophilic index of 90%. Of the 25 specimens from cattlesheds 15 were positive for human blood with an index of 60%. Of the 20 specimens from mixed dwellings 12 showed human blood with an anthropophilic index of 63.15%. Thus, the Culex quinquefasciatus at Raipur city in Madhya Pradesh state is predominantly anthropophilic in nature irrespective of the nature of habitat of the mosquito vector. PMID- 11898459 TI - Detection of antibodies to Taenia solium in sera of patient with epilepsy using ELISA. AB - Enzyme linked immunosorbent assay (ELISA) was used for detection of antibodies to cysticerci of Taenia solium in patients suffering from suspected neurocysticercosis. Serum antibodies to cysticercosis were detected in 10.4% of the patients. Antibodies were absent in healthy controls. No cross-reactivity was observed in sera from patients with other parasitic diseases viz. toxoplasmosis, filariasis and kala-azar. However sera from patients with hydatid disease showed cross reactions in 20%. Age group distribution showed gradual non-significant increase in seropositivity rates with advancing age. No significant gender difference, was observed. Analysis of dietary habits of patients showed statistically significant difference in seropositivity rates in non-vegetarians (15.4%) as compared to vegetarians (6.9%) (P value < .001). PMID- 11898460 TI - Observations on the breeding habitat of Aedes species in the steel township, Rourkela. AB - Prompted by report of large number of dengue fever cases in township of Rourkela steel plant, entomological surveys were carried out to know the distribution and extent of Aedes breeding in and around the dengue fever affected areas. Out of 2062 water containers searched, 819 were positive for Aedes larvae. The house index (HI), container index (CI) and Breteau index (BI) were 53.4, 39.7 and 118.5 respectively. 27.9% of the houses had single breeding habitats. The breateau index of single storeyed houses and double storeyed buildings was 149.8 and 54.6 respectively, showing that the single storeyed houses have more potential for Aedes breeding. The breeding preference ratio (BPR) was highest for plastic containers. Three species of Aedes were found breeding in different habitats and Aedes aegypti was found breeding in coolers, cement tanks, tyres and miscellaneous containers with a prevalence rate of 13.2%. The role of entomological surveillance and health education has been highlighted for the control of disease through community participation. PMID- 11898461 TI - Serological diagnosis of human brucellosis: analysis of seven cases with neurological and cardiological manifestations. AB - Serum Tube Agglutination (STA) test was used as routine test to detect antibrucellar antibodies in diagnosis of brucella infection in sera (n = 75) and CSF (n = 14) from 78 patients with neurological (n = 60) and cardiological (n = 15) complaints in whom brucellosis was suspected, over a period of two and a half years from January, 1997 to July 1999. Seven (neurological-six and cardiac-one) serum samples (9.33%) were positive by STA, while none of the CSFs were positive. STA titres ranged from 1:10 to 1:1280. We report the findings of these seven cases with neurological and cardiac manifestations in whom STA were found positive. Treatment was accomplished in two cases (neurological-one and cardiac one), while remaining five cases either were treated empirically with antitubercular treatment or lost for follow up. However these reported cases should alert clinicians to investigate for Brucella infection in cases of pyrexia of unknown origin and this condition in cases of chronic meningitis with unproven aetiology. PMID- 11898462 TI - Shima drinking water--a bacteriological analysis. AB - 300 water samples, 60 from piped supply and 240 from 20 natural sources were analyzed bacteriologically for four important bacterial indicators every month over a period of one year. The MPN of total coliforms, Escherichia coli, Streptococcus faecalis were detected by multiple tube method and Clostridium perfringens was isolated using litmus milk medium. From piped water supply, only one sample was found unfit for human consumption. The plate count at 37 degrees C and 22 degrees C varied from 0.5 x 10(3) to 15 x 10(3) per ml. of water. From natural sources, MPN indices for coliforms, Esch. coli and S. faecalis varied from 3 to > 1800, 0 to > 1800 and 0 to 540 per 100 ml. of water respectively. About 1/3rd of samples showed presence of Cl. perfringens. The plate count varied from 1.0 x 10(3) to > 150 x 10(3) per ml. of water. Salmonella typhi was isolated in 1.25% samples by membrane filtration technique. The water from all the natural sources was unfit for human consumption whereas piped water supply was of good quality in general. PMID- 11898463 TI - Man biting activity of filarial vector Culex quinquefasciatus. PMID- 11898464 TI - Sandfly survey in Nainital and Almora districts of Uttaranchal with particular reference to Phlebotomus argentipes, vector of kala-azar. AB - Kala-azar continues to pose a major public health problem in Bihar, West Bengal and parts of eastern Uttar Pradesh in India causing great deal of morbidity and mortality. During 1998, several kala-azar cases from Sub-Himalayan region were treated in Delhi hospitals. And a suspected focus of kala-azar was subsequently reported from this area. Therefore a preliminary sandfly survey was carried out during October, 1999 in 18 randomly selected villages at different altitudes in Nainital & Almora districts of Kumaon region Uttaranchal. The surveys revealed relative preponderance of vector sandfly Ph. argentipes as 77%; mainly confined to cattlesheds and mixed dwellings in the villages at an altitude ranging from 350-960 metres main sea level. The other sandfly species encountered were: Ph. papatasi 6.9%, Ph. major 2.9% and 13.2% Sergentomyia spp. 17.4% Ph. argentipes reacted positive with human antisera and 82.6% with bovine but none reacted with goat, pig dog and bird antisera indicating that Ph. argentipes in the area is mainly zoophilic. Ph. argentipes was found to be highly susceptible to DDT; mortality ranging from 98-100%. PMID- 11898465 TI - Correlation of clinical, histological and immunological features across the leprosy spectrum. AB - The Ridley-Jopling system of classification of the variegated clinical pattern of leprosy is based on the specific cell-mediated immunity observed in the histopathology of skin lesions conforming to a spectrum from TT at one end to LL at the other. In this study a fairly large sample of 90 patients was classified on clinical grounds; the histopathology of the skin lesions was studied blind. There was an overall concordance of 90% between the clinical and histological classifications. In addition, the systemic cell-mediated and humoral immune responses were studied. The in vivo cell-mediated immune response, namely the Mitsuda skin response, mostly conformed to the clinical classification. While the in vitro lymphoproliferative responses to BCG and its sonicate were high, the lymphoproliferative responses to Dharmendra lepromin were surprisingly poor. Humoral responses to 35 kDA protein of M. leprae and PGL-1 were good in most LL, BL patients and tapered off towards TT. IgG antibodies to recombinant ML 65 kDa proteins denoted mycobacterial presence. PMID- 11898466 TI - Localized linear lesions in lepromatous leprosy. PMID- 11898467 TI - Smear positive leprosy with HIV infection, silent neuritis and extensive tinea corporis and tinea unguium in an Indian male. PMID- 11898468 TI - Histoid leprosy masquerading as tuberous xanthomas. PMID- 11898469 TI - Pure neuritic leprosy of supra-orbital nerve--as unusual presentation. PMID- 11898470 TI - Indeterminate leprosy, molluscum contagiosum and nevus depigmentosus. PMID- 11898471 TI - Relapse rates amongst the multidrug therapy-treated leprosy patients. PMID- 11898472 TI - A comparative trial of single-dose chemotherapy in paucibacillary leprosy patients with two to three skin lesions. PMID- 11898473 TI - President's address. Leprosy in India--looking back and looking forward. PMID- 11898475 TI - Tricyclic antidepressant medication, stress management therapy, and their combination in the management of chronic tension-type headache. PMID- 11898476 TI - Hemicrania continua: recent treatment strategies and diagnostic evaluation. AB - Hemicrania continua (HC) is a primary headache disorder that is characterized by a continuous unilateral headache of moderate severity, exacerbations of severe pain, and complete responsiveness to indomethacin. HC was once thought to be a rare headache disorder, but now many cases have been reported. It is an underecognized headache syndrome. HC can be of continuous or remitting form. Variants such as hemicrania continua with aura have been described, and secondary cases may occur. Indomethacin is the best treatment, although HC could respond to other nonsteroidal anti-inflammatory drugs, such as the selective cyclo-oxygenase 2 inhibitors. PMID- 11898477 TI - New treatments in cluster headache. AB - Cluster headache is the most severe headache syndrome known to humans. In most instances, this disorder is readily treatable when the correct medications are utilized at the correct dosages. Cluster treatment involves abortive, transitional, and preventive therapy strategies, all of which are discussed in this article. PMID- 11898479 TI - Radiation-induced neurocognitive decline: the risks and benefits of reducing the amount of whole-brain irradiation. PMID- 11898478 TI - Modular headache theory: a new approach. AB - Many people experience headaches that do not fulfill the International Headache Society's criteria for a specific headache disorder, yet behave biologically like that disorder. Others fulfill criteria for one headache disorder but have features of another. To explain these observations, we propose that groups of neurons, called modules, become activated to produce each symptom of a primary headache disorder, and that each module is linked to other modules that together produce an individual's headache. Headaches develop phenotypic stability through a process referred to as learned stereotypy. This theory has implications for the classification, research, and treatment of primary and secondary headache patients. PMID- 11898480 TI - New antiepileptic drugs: comparative studies of efficacy and cognition. AB - Over the past 10 years, the Food and Drug Administration has approved eight prophylactic antiepileptic drugs (AEDs). Although the goal of universal seizure control without side effects has not been reached, the tolerability of medications has improved. This paper reviews the AEDs introduced since 1993 and tries to objectively present comparative data obtained in double-blind studies. PMID- 11898481 TI - The utility of functional magnetic resonance imaging in epilepsy and language. AB - Functional magnetic resonance imaging (fMRI) is a viable presurgical tool for use with the pediatric epilepsy population as replacement for the intra-carotid sodium amobarbital test (IAT) used to identify hemispheric language dominance. This paper reviews the current imaging research on the identification of language cortex in pediatric epilepsy patients and in normal children. A review of the literature comparing fMRI to the IAT and electrocortical stimulation suggests that fMRI reliably identifies the dominant hemisphere, with pediatric and adult studies producing comparable results. Within-hemisphere localization of eloquent cortex with fMRI is more problematic. Paradigm selection, data analysis techniques, and considerations specific to imaging children are discussed. Utility of fMRI for studying neural plasticity as a result of brain insult (e.g., epilepsy) is also considered. PMID- 11898482 TI - Childhood sleep disorders: diagnostic and therapeutic approaches. AB - Pediatric sleep physiology begins with development of the sleep/wake cycle, and the origins of active versus quiet sleep. The 24-hour circadian cycle becomes established at 3 to 6 months. Sleep disorders are rationally approached in pediatrics as age-related. Disorders during infancy commonly include mild, usually self-limited conditions such as sleep-onset association disorder, excessive nighttime feedings, and poor limit-setting. These require behavioral management to avoid long-term deleterious sleep habits. In contrast, other sleep disorders are more ominous, including sudden infant death syndrome (SIDS), central congenital hypoventilation syndrome, and sleep apnea. Childhood is generally the golden age of sleep, with brief latency, high efficiency, and easy awakening. Parasomnias, sometimes stage specific, are manifest here. Adolescents have sleep requirements similar to preteens, posing a challenge for them to adapt to school schedules and lifestyles. Narcolepsy, usually diagnosed in adolescence or early adulthood, is a lifelong sleep disorder that has led to the identification of the hypocretin/orexin neurotransmitter system. This will lead to enhanced understanding of what regulates stage rapid eye movement, and to novel therapeutic advances for hypersomnolence. PMID- 11898483 TI - Neurologic manifestations of tuberous sclerosis complex. AB - Tuberous sclerosis complex (TSC) is a multiorgan disorder that primarily affects the brain, skin, and kidneys. Recent advances have elucidated the genetics of this complex, which has helped lead to an increased understanding of the basic neurobiology of this disorder. There is both phenotypic and geneotypic heterogeneity. The treatment of epilepsy remains a major challenge in these patients, and there is an increasing role for epileptic surgery. Many patients with TSC continue to have intractable seizures. Early identification to ensure proper monitoring and genetic counseling continue to be important clinically. The neurologist must be aware of other organ involvement, particularly the kidneys, and the lungs in female patients, to ensure appropriate monitoring for complications. It is also important to be aware of the marked variability of expression in all the clinical features of TSC. PMID- 11898484 TI - Current management of sleep disturbances in dementia. AB - The management of sleep disturbances in patients with dementia is a complicated and enormously important clinical and societal problem. In this review, we present one approach to the diagnosis and management of such sleep disturbances. Most disturbances can be categorized into four primary symptoms: insomnia, hypersomnia, excessive nocturnal motor activity, and hallucinations or behavioral problems. We describe how each symptom may relate to the dementing illnesses themselves, which primary sleep disorders may be at play, which medications employed for dementia may impact on the symptom, the role of depression in that symptom, and how circadian dysrhythmias can underlie that symptom. Although few well-designed studies have been conducted, we present management strategies for several sleep disturbances based on the literature and our clinical experience. Considering the impact on patient and caregiver quality of life, and the potential for delaying institutionalization with appropriate therapy, further research is clearly warranted to optimize the diagnosis and management of sleep disturbances in the cognitively impaired elderly population. PMID- 11898485 TI - Sleep changes and disorders in the elderly patient. AB - Sleep disorders are very common among older patients. Our population as a whole is experiencing substantial growth in the geriatric population. Thus, it is all the more prudent that healthcare professionals become familiar with the major sleep disorders that affect the older patient, as well as the diagnosis and treatment of these disorders. This paper deals with the various sleep changes that take place in the course of the aging process and with the assessment of sleep disorders in the older patient. Special attention is paid to sleep disordered breathing, periodic limb-movement disorder of sleep, insomnia, and circadian rhythm anthology, as well as parasomnias that are unique to the older patient. The final section of the paper deals with specific neurologic disorders and their impact on sleep patterns. PMID- 11898487 TI - Risks of bovine spongiform encephalopathy and variant Creutzfeldt-Jakob disease in the United States. PMID- 11898488 TI - Evolution in holistic nursing. PMID- 11898489 TI - Talking about human subjects. PMID- 11898486 TI - Restless legs syndrome: a sensorimotor disorder of sleep/wake motor regulation. AB - Restless legs syndrome (RLS) remains an underappreciated sensorimotor disorder of sleep/wake regulation. It is one of the few sensorimotor disorders that is provoked by rest and that also follows a clear circadian pattern. Recent epidemiologic studies have verified that the condition is common in populations derived from the north and west of Europe, and have begun to uncover some of the genetic substrate of the disorder. New instruments have been developed to facilitate diagnosis and assessment of severity. The pathogenesis of the condition remains uncertain, but recent discoveries implicate areas of the nervous system from the spinal cord up to the basal ganglia. A current hypothesis undergoing vigorous exploration is that the condition results from a deficiency of dopaminergic function based on abnormalities of iron transport and storage. Therapeutically, studies have shown the dopamine agonists to be the most reliable treatment for severe cases, whereas other recent studies have successfully utilized a number of other medications, including levodopa, opioids, and anticonvulsants. New standards provide guidelines for management of RLS and make specific pharmacotherapeutic recommendations. PMID- 11898490 TI - Educating our state boards of nursing on holistic nursing practice--Part II. PMID- 11898492 TI - A holistic health care 'bill of rights'. PMID- 11898491 TI - Nightingale moments. PMID- 11898493 TI - Nobels, neurologists, and Moore's law. PMID- 11898494 TI - Antiplatelet aggregating versus anticoagulant agents in preventing early recurrent stroke among patients with atrial fibrillation. PMID- 11898495 TI - Current status of neuroprotective agents in the treatment of acute ischemic stroke. AB - To keep ischemic brain cells alive, neuroprotective agents target events in the ischemic cascade that might be injurious to the cells. They can be divided broadly into groups that restore ion balance, block receptors, prevent reperfusion injury, or promote neuronal healing. To date, neuroprotective agents have either shown a lack of efficacy in clinical stroke trials or been limited by side effects. Ongoing clinical trials with novel agents are trying to enroll a more homogeneous population of stroke patients in an effort to demonstrate treatment benefit. PMID- 11898496 TI - New insights on thrombolytic treatment of acute ischemic stroke. AB - In 1995, a two-part randomized trial showed the efficacy of intravenous tissue plasminogen activator (tPA) when given within 3 hours of onset of symptoms of acute ischemic stroke. Two subsequent trials were unable to extend the therapeutic window of intravenous tPA beyond 3 hours. A phase IV study performed by experienced stroke centers showed an acceptably low symptomatic intracerebral hemorrhage rate for intravenous tPA of only 3%, whereas a review of the Cleveland area experience showed a disturbingly high rate of symptomatic intracerebral hemorrhage of 15.7%. The Prolyse in Acute Cerebral Thromboembolism (PROACT) II study showed efficacy of intra-arterial pro-urokinase and intravenous heparin over intravenous heparin alone when given within 6 hours of onset of symptoms to patients with thrombotic occlusion of the proximal middle cerebral artery. Additional controlled investigations of intra-arterial thrombolytic therapy are needed. Neuroprotectants in combination with intravenous tPA have yet to show improved efficacy over the use of tPA alone. PMID- 11898498 TI - Choices in medical management for prevention of acute ischemic stroke. AB - Stroke is a leading cause of death and disability. Although advances are being made in the treatment of acute ischemic stroke, its prevention is equally as important. Identification and management of risk factors are essential. Medical therapy is also helpful in the secondary prevention of ischemic stroke. There are currently four platelet-antiaggregating agents used to prevent ischemic stroke: aspirin, aspirin plus dipyridamole, clopidogrel, and ticlopidine. The relevant studies proving their efficacy are noted, as are some of their similarities and differences. The use of warfarin is also discussed. PMID- 11898497 TI - The role of neuroimaging in selecting treatments for patients with acute stroke. AB - The armamentarium available for treating acute stroke patients is growing as clinical trials show benefit of intravenous thrombolysis, intra-arterial clot lysis, and antiplatelet agents. Patients with dissection or severe atheromatous stenosis of major cerebrovascular vessels are commonly treated with anticoagulation to prevent recurrent artery-to-artery embolus or arterial thrombosis. These advances in acute stroke treatments demand an accurate means to quickly identify those patients most likely to benefit (or not benefit) from a specific therapy. Fortunately, advances in imaging cerebrovascular lesions, decreased brain perfusion, and even ischemic tissue injury now make it possible to consider tailoring therapy to the individual patient's cerebrovascular problem. Experience and controlled clinical data in this endeavor is meager. Here we describe the ability of various emergency neuroimaging tools to provide information on the state of brain blood flow, metabolism, and vascular anatomy. Most importantly, we present the rationale and limited available evidence relevant to how the neuroimaging information might be used to select optimal treatments for individual patients. PMID- 11898500 TI - Literature alert. PMID- 11898499 TI - Cerebrovascular angioplasty and stenting for the prevention of stroke. AB - Until recently, stroke preventive strategies have focused on either medical regimens aimed at antagonizing or reversing atherosclerosis, or surgical intervention for revascularization of the cerebrovascular system. However, with the advent of rapidly emerging microcatheterization techniques and technology, endovascular surgical revascularization of the brain is rapidly emerging as a powerful therapeutic modality. In particular, significant advances already have been made in revascularization of the extracranial carotid artery and many common anatomic sites of intracranial athero-occlusive disease, using special adaptations of conventional percutaneous angioplasty and stenting techniques. This paper reviews the cumulative experience with these emerging techniques, with a particular emphasis on clinical outcomes and future directions. It also reports the substantial cumulative institutional experience of the authors over the past 18 months with both extracranial carotid and intracranial artery stent-assisted carotid angioplasty. PMID- 11898501 TI - Stroke in young adults and children. AB - Data from studies of 337 children and 1606 young adults are summarized to identify the major causes of stroke in these age groups. In children under 15 years of age, stroke occurs in patients with congenital heart disease, nonatherosclerotic vasculopathies, infection, and hematologic defects like sickle cell disease. In patients 15 to 35 years of age, dissection, cardioembolism, nonatheroslerotic vasculopathies, and prothrombotic states cause a significant percentage of strokes. In adults over 35 years of age, traditional atherosclerotic risk factors predominate. Lifestyle choices (e.g., cigarette smoking, alcohol consumption, and illicit drug use) can significantly increase the rate of stroke among young adults in a community. Limited access to healthcare may increase the role of infectious disease and peripartum complications. PMID- 11898502 TI - Progress in the pathogenesis of amyotrophic lateral sclerosis. AB - This decade has seen the discovery of one cause for amyotrophic lateral sclerosis (ALS)--mutations in the copper/zinc superoxide dismutase (SOD1) gene. Mutant SOD1 has provided an invaluable tool for transgenic and cellular experiments designed to elicit the biochemical pathways that are disturbed in ALS. We highlight recent advances in ALS research, including diagnostic issues, new loci for ALS genes, and progress in understanding the toxicity of mutant SOD1. The evidence for persistant viral infection, glutamate-mediated excitotoxicity, oxidative stress, altered neurofilament and peripherin expression, disrupted axonal transport, neurotrophin deficiency, and mitochondrial dysfunction are critically reviewed. As yet, no consensus has been achieved on the pathways that lead to selective neuronal death, and the underlying causes are still unknown in the vast majority of patients. Further clues about genetic susceptibility and environmental triggers are urgently needed so that more effective treatments for ALS can be developed, with the ultimate goal being prevention. PMID- 11898503 TI - A molecular basis for hereditary motor and sensory neuropathy disorders. AB - Charcot-Marie-Tooth disease (CMT), or inherited peripheral neuropathies, is one of the most frequent genetically inherited neurologic disorders, with a prevalence of approximately one in 2500 people. CMT is usually inherited in an autosomal dominant fashion, although X-linked and recessive forms of CMT also exist. Over the past several years, considerable progress has been made toward understanding the genetic causes of many of the most frequent forms of CMT, particularly those caused by mutations in Schwann cell genes inducing the demyelinating forms of CMT, also known as CMT1. Because the genetic cause of these disorders is known, it is now possible to study how mutations in genes encoding myelin proteins cause neuropathy. Identifying these mechanisms will be important both for understanding demyelination and for developing future treatments for CMT. PMID- 11898505 TI - Literature alert. PMID- 11898504 TI - Progress in gene therapy for Duchenne muscular dystrophy. AB - Gene transfer research for Duchenne muscular dystrophy (DMD) has brought the goal of successful treatment of this devastating, inherited disease closer to being a reality. Although gene therapeutic approaches for DMD patients are not yet in clinical use, recent advances using DMD animal models are encouraging. Progress in vector design, such as high-capacity adenoviral vectors, targeted adenoviral vectors, and heterodimerization of DNA delivered by adeno-associated virus (AAV) vectors have advanced the field considerably. The recent studies into the pharmacologic-induced read-through of stop codons, the increased study of utrophin and its upregulation, and the introduction of point mutation correction using chimeric oligonucleotides have expanded the field, providing new avenues of inquiry. PMID- 11898506 TI - Spontaneous intracranial hypotension. AB - Spontaneous intracranial hypotension (SIH) is typically manifested by orthostatic headaches that may be associated with one or more of several other symptoms, including pain or stiffness of the neck, nausea, emesis, horizontal diplopia, dizziness, change in hearing, visual blurring or visual field cuts, photophobia, interscapular pain, and occasionally face numbness or weakness or radicular upper limb symptoms. Cerebrospinal fluid (CSF) pressures, by definition, are quite low. SIH almost invariably results from a spontaneous CSF leak. Only very infrequently is this leak at the skull base (cribriform plate). In the overwhelming majority of patients, the leak is at the level of the spine, particularly the thoracic spine and cervicothoracic junction. Sometimes, documented leaks and typical clinical and imaging findings of SIH are associated with CSF pressures that are consistently within limits of normal. Magnetic resonance imaging of the head typically shows diffuse pachymeningeal gadolinium enhancement, often with imaging evidence of sinking of the brain, and less frequently with subdural fluid collections, engorged cerebral venous sinuses, enlarged pituitary gland, or decreased size of the ventricles. Radioisotope cisternography typically shows absence of activity over the cerebral convexities, even at 24 or 48 hours, and early appearance of activity in the kidneys and urinary bladder, and may sometimes reveal the level of the leak. Although various treatment modalities have been implemented, epidural blood patch is probably the treatment of choice in patients who have failed an initial trial of conservative management. When adequate trials of epidural blood patches fail, surgery can offer encouraging results in selected cases in which the site of the leak has been identified. Some of the spontaneous CSF leaks are related to weakness of the meningeal sac, likely in connection with a connective tissue abnormality. PMID- 11898507 TI - Chronic daily headache. AB - Chronic daily headache (CDH) is a heterogeneous group of headaches that includes primary and secondary varieties. Primary CDH is a frequent entity that probably affects 4% to 5% of the population. It can be subdivided into headaches of short duration (less than 4 hours per attack), like chronic cluster headache, and disorders of long duration (greater than 4 hours per attack). Primary CDH of long duration includes transformed migraine, chronic tension-type headache, new daily persistent headache, and hemicrania continua. Analgesic or ergot overuse is frequent in all types of CDH. We review recent insights into the proposed classification, epidemiology, pathophysiology, clinical characteristics, and treatment of CDH. PMID- 11898508 TI - Acute treatment of migraine and the role of triptans. AB - The use of triptans has improved the ability to treat migraine successfully compared with older treatments. Speed of relief, consistency of effect, and good tolerability have been the hallmarks of these agents. All of the currently available triptans have comparable efficacy and tolerability. Variables between the agents may lead to one agent or dose form being preferred over another in various clinical scenarios. The triptans that are forthcoming may improve on these options through enhanced efficacy rates, tolerability, and headache recurrence rates. There exist increasing options for migraine treatment that may further improve the clinical effects of the older and newer triptans through early treatment of migraine at the stages of mild migraine pain, or even during the prodromal phase of the attack. Additionally, recent work suggests that mini prophylaxis of migraine at the menses is a highly successful treatment option with the triptans. In this age of managed care, providing cost-effective treatment of headache will take on increasing importance. Techniques such as stratification of acute treatments may enhance cost-effective care, whereas ready availability of the triptans may lead to significant improvements in utilization of parameters such as office visits, emergency room treatment, and even hospitalization. PMID- 11898509 TI - Advances in biology and treatment of childhood brain tumors. AB - Childhood brain tumors are collectively the most common solid neoplasm and the leading cause of cancer-related death in children. They are a diverse group of diseases and outcome is extremely variable. Current treatment is dependent on histology, location, and in some instances, patient age. Advances in treatment have led to improved survival for some patients, but for many the outcome remains dismal despite aggressive treatment. A growing body of work is aimed at improving the outcome for children with brain tumors not only through clinical trials, but also by focusing on the biologic underpinning of these diseases that have been poorly understood. PMID- 11898510 TI - Treatment of childhood headaches. AB - Migraine and tension-type headaches are two of the most common types of primary headache disorders in children. Migraine is a primary central nervous system disorder characterized by triggered or spontaneous episodes of activation of trigemino-vascular complex, neurogenic inflammation around vessels and meninges, and stimulation of the peripheral and central pain pathways of the trigemino cervical complex. The triptans, by their selective agonistic action on 5-HT1B/1D receptors, are very effective in the treatment of migraine pain and associated symptoms. Early studies on the safety and efficacy of triptans in the management of childhood migraine show encouraging results. We propose a stratified-care model for the management of migraine in children, and discuss pharmacotherapy based on the pathophysiologic mechanisms of migraine pain. Management of tension type headaches requires comprehensive medical and psychologic evaluation and an individualized approach for a successful outcome. PMID- 11898511 TI - The anticonvulsant effect of electrical fields. AB - The use of electrical fields to treat epilepsy is undergoing increased scrutiny as an alternative to medications and resective surgery. Much recent attention has been focused on ionic channels and seizure control; however, nonsynaptic mechanisms may be crucial for seizure onset, raising the possibility of using electrical field application to abort seizures. Furthermore, the inhibitory effects may outlast the immediate treatment and possibly be a prophylactic intervention. This paper reviews the use of brain stimulation for treatment of epilepsy, but also cites instances where the antithetical results occur. The greatest detail focuses on disrupting the onset or shortening the seizure. The paper does not extensively review deep brain or vagal nerve stimulation. PMID- 11898512 TI - Diagnosis and management of neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1) is an autosomal dominant disorder whose major feature is the occurrence of multiple neurofibromas, which are benign tumors of the nerve sheath. It affects an estimated one in 3000 to 4000 individuals. In addition to neurofibromas, there are many other clinical manifestations, including malignant tumors such as gliomas or malignant peripheral nerve sheath tumors, and nontumor effects such as skeletal dysplasia and learning disability. Diagnosis is established on the basis of clinical criteria. Molecular genetic testing is feasible, but the large size of the gene and wide range of pathogenic mutations have so far impeded the development of a clinical diagnostic test. Insights into pathogenesis have followed from identification of the NF1 gene and the development of animal models. The major function of the gene product appears to be regulation of the ras protein. Tumors are believed to arise by the loss of function of the NF1 protein, suggesting that NF1 behaves as a tumor suppressor gene. Heterozygous effects on some cell types are also likely, however. The role of ras in the pathogenesis of tumors in NF1 has suggested an approach to treatment using ras inhibitors, some of which are likely to begin in clinical trials in NF1 patients in the near future. PMID- 11898513 TI - Pharmacologic and genetic therapy for childhood muscular dystrophies. AB - The outstanding advances in the molecular characterization of muscle diseases, including muscular dystrophies, inflammatory myopathies, and ion channel disorders, have resulted in the identification of potential targets for pharmacologic and genetic therapy in the best characterized of these diseases. The most common myopathy in children, Duchenne muscular dystrophy (DMD), is the focus of active pharmacologic clinical trials. Genetic transfer therapy research for this and other dystrophies is rapidly moving forward. However, as new approaches for treatment are being actively investigated, the current modality of treatment for all myopathies is still in the realm of physical medicine and rehabilitation. The focus of this review is on the advances in pharmacologic and genetic therapy research in DMD and limb girdle muscular dystrophies. PMID- 11898514 TI - Perinatal asphyxia: timing and mechanisms of injury in neonatal encephalopathy. AB - This article summarizes the recent medical literature regarding perinatal asphyxia with respect to timing and mechanisms of injury for neonates who were clinically diagnosed with an encephalopathy in the newborn period. Multiple mechanisms of injury are reviewed, including genetic vulnerability, acquired inflammatory responses, and clotting defects that can lead to ischemic-induced brain damage. Before effective treatments for fetal and neonatal brain disorders can be developed, accurate and timely diagnoses of fetal or neonatal brain injury must be achieved. Specific subsets of children can then benefit from neuroprotective strategies that can target the specific developmental aspects of brain adaptation or plasticity relative to the specific etiology and timing of injury after asphyxia. PMID- 11898515 TI - Diagnosis and treatment of childhood mitochondrial diseases. AB - Mitochondrial cytopathies are caused by genetic alterations of nuclear- or mitochondrial-encoded genes involved in the synthesis of subunits of the electron transport chain. Mutations of mitochondrial DNA are associated with a wide range of clinical presentations [1-4]. The ubiquitous nature of mitochondria and the role of the mitochondria in cellular metabolism result in the potential for any tissue in the body to be affected [5-7,8..,9]. Although some children with mitochondrial disease present with life-threatening lactic acidosis in the newborn period, the majority of children come to clinical attention for nonspecific problems, including failure to thrive, developmental delay, seizures, hypotonia, and loss of developmental milestones. The diagnosis of these disorders is made through careful clinical evaluation, coupled with biochemical, morphologic, and molecular biologic techniques. Genetic counseling is difficult due to unique aspects of mitochondrial genetics. Despite advances in our understanding of mitochondrial biochemistry and genetics, treatment options remain limited. PMID- 11898518 TI - Cerebral energy failure. PMID- 11898517 TI - Facts and an open mind should guide clinical practice. PMID- 11898516 TI - The treatment of tics. AB - Tics are the essential component of a variety of disorders, notably Tourette syndrome. Because the mere presence of tics does not require therapeutic intervention, criteria are essential for determining a functional disability. Suggested tic-suppressing treatments have been extremely diverse, including hydrotherapy, behavioral treatments, pharmacotherapy, botulinum toxin, immunomodulatory therapy, and surgery. This manuscript reviews each of these approaches with emphasis on more recent pharmacologic trials. PMID- 11898519 TI - Literature alert. PMID- 11898520 TI - Drug delivery to tumors of the central nervous system. AB - Contemporary treatment of malignant brain tumors has been hampered by problems with drug delivery to the tumor bed. Inherent boundaries of the central nervous system, such as the blood-brain barrier or the blood-cerebrospinal fluid barrier, and a general lack of response to many chemotherapeutic agents have led to alternative treatment modalities. In general, all these modalities have sought to either disrupt or bypass the physiologic brain barriers and deliver the drug directly to the tumor. This article reviews past, as well as current, methods of drug delivery to tumors of the central nervous system. Special emphasis is placed on biodegradable polymers that can release chemotherapeutic agents against malignant gliomas. A variety of other nonchemotherapeutic drugs, including antiangiogenesis and immunotherapeutic agents, are presented in the context of new polymer technology. Finally, future directions in drug delivery are discussed with an overview on new advances in emerging biotechnology. PMID- 11898522 TI - Migration and invasion in brain neoplasms. AB - Local invasion of the brain by neoplastic glial cells is a major obstacle to effective treatment of intrinsic brain tumors. Invasion is directly related to histologic malignancy, but occurs to some extent irrespective of tumor grade. Because the brain-to-tumor interface is not well demarcated, total surgical removal is rarely possible; moreover, as invading cells transiently arrest from cell division they are refractory to radiotherapeutic intervention. Invading cells may also be protected from the action of cytotoxic drugs by the presence of an intact blood-brain barrier. The invading cells, having migrated several millimeters or even centimeters from the main focus of the tumor, return to cycle phase under the control of some as yet unknown microenvironmental cue to form a recurrent tumor adjacent to the original site of presentation. Recent cellular and genetic information concerning factors underlying invasion may not only yield suitable targets for adaptation of existing therapies, but may also lead to novel approaches in glioma management. PMID- 11898521 TI - Genetics of brain neoplasms. AB - Transformation of a normal cell into a malignant cell involves a series of events that damage the genome. Gliomas are the most common adult neoplasm of the central nervous system. To develop new therapeutic strategies requires an understanding of the specific lesions that occur and contribute to this malignant process. Initially, data reported from the analyses of human gliomas were quite variable. This has recently changed as more data have become available and the selection of tissue analyzed is coupled with clinical criteria. Specific genetic lesions are now defining different glioma pathways, and some aberrations may be indicative of therapeutic response. This review focuses on the specific genetic aberrations associated with astrocytic and oligodenroglial tumors. PMID- 11898523 TI - Advances in stereotactic radiosurgery for brain neoplasms. AB - It has been nearly half a century since Leksell introduced brain radiosurgery. In the past decade, the procedure has become widely used as both a primary and adjuvant treatment. Radiosurgery is now commonly employed for arteriovenous malformations, brain metastases, and several benign lesions. Its use in primary brain malignancy remains of uncertain benefit. Improvements in imaging, hardware, and software promise an even greater role for the technique. PMID- 11898524 TI - Immunologic approaches to therapy for brain tumors. AB - Malignant brain tumors are notoriously invasive. Although surgical debulking can relieve the patient of the main mass of tumor, adjuvant treatments are needed to target the glioma cells that infiltrate through normal parenchyma as single cells or pockets of tumor cells from which recurrent tumors arise. Successful adjuvant cellular therapy of brain tumors, or activation of endogenous immune cells, requires that either cell effectors make direct contact with tumor cells or come within close proximity to them and exert an indirect effect. This review examines current clinical trials aimed at direct lysis of glioma cells and trials making gliomas more visible to the endogenous immune system. PMID- 11898526 TI - Spectrum and classification of inflammatory demyelinating diseases of the central nervous system. AB - The aim of this review is to highlight recent observations concerning the pathogenesis of acute and chronic inflammatory demyelinating diseases in the central nervous system. Without attempting to provide a didactic classification or a complete survey, we emphasize the discriminative nature of new clinical, imaging, and immunopathologic data, which, even in the absence of specific molecular markers, modify our views about the nosologic relations among these overlapping clinicopathologic entities. In the light of new findings, multiple sclerosis may represent a spectrum of demyelinating diseases rather than a single entity. PMID- 11898527 TI - Immunopathogenesis of the multiple sclerosis lesion. AB - Multiple sclerosis (MS) is thought to be an autoimmune disease with a chronic inflammatory response directed against central nervous system (CNS) myelin antigens. Immunologic studies indicate that autoreactive CD4+ lymphocytes migrate into the CNS causing blood brain barrier (BBB) disruption, an initial event in the evolution of the MS lesion. Subsequent antigen recognition within the CNS initiates inflammatory responses that, through the multiple effector mechanisms, lead to demyelination. Magnetic resonance imaging (MRI) studies provide new insights into the evolution of the MS lesion, revealing an active and continuous pathologic process that is not only localized to focal lesions, but also diffusely affects normal appearing white matter (NAWM). Standard T2-weighted images are exquisitely sensitive, showing changes due to inflammation, edema, demyelination, and axonal loss, but because of the lack of pathologic specificity, they only moderately correlate with the clinical parameters. New MRI techniques, including magnetic resonance spectroscopy, magnetization transfer, and diffusion imaging, provide a better measure of axonal loss and demyelination, the most clinically relevant components of MS lesions. Hopefully, they will enable us to more accurately monitor disease activity and evaluate the effects of new therapies on the progression of the disease. PMID- 11898528 TI - T cells, cytokines, and autoantigens in multiple sclerosis. AB - In multiple sclerosis (MS), inflammatory demyelination in the central nervous system is thought to be initiated by T cells that recognize myelin antigens. T cells are the main regulators of acquired immunity and are involved in the pathogenesis of several organ-specific autoimmune diseases. This review provides an overview of recent studies on the role of T cells in autoimmune demyelination. Because autoreactive T cells are normally present in the mature repertoire of T cells in the blood and lymphoid organs of MS patients, but also in normal controls, particular attention is devoted to the mechanisms of activation and the functional phenotype of such T cells in patients with MS. The role of cytokines as effector molecules and the main candidate antigens are also discussed. PMID- 11898529 TI - Infection and the etiology and pathogenesis of multiple sclerosis. AB - Multiple sclerosis (MS) currently defies clinical and scientific definitions, and carries a prognosis that remains practically unchanged despite many years of intensive research. Although the prevailing dogma is that MS is an immune mediated condition, it fulfills none of the criteria of an autoimmune disease. On the other hand, there is enough significant data to suggest that infectious agents(s) could be involved in either direct damage to the white matter or induce inflammatory responses that secondarily affect the brain. Our goal here is to review the data supporting the possibility that infection has a critical role in the disease, examine the list of potential candidates that have been suggested, and outline an approach regarding the potential role of infectious agents in the etiology and pathogenesis of MS. PMID- 11898530 TI - Immunologic therapy for relapsing-remitting multiple sclerosis. AB - The treatment of relapsing-remitting multiple sclerosis (RRMS), the most common form of multiple sclerosis, has been revolutionized in recent years. In addition to effective treatment of acute relapses, therapies are now available to prevent relapses, reduce the burden of disease as seen on magnetic resonance imaging, and possibly even slow the course of disease. There are now several agents either approved, awaiting approval, or in various stages of development in many countries. Evidence suggests that early intervention with these agents will yield the best results in the long run. The current approach to treatment of RRMS is the focus of this discussion. PMID- 11898532 TI - Fatigue in multiple sclerosis. AB - Fatigue is among the most common, yet least understood, symptoms of multiple sclerosis (MS) [1.]. It can profoundly disrupt the occupational and social functioning of patients, and is recognized as a criterion for MS disability by the Social Security Administration. Most approaches to fatigue assessment can be classified as either self-report scales or performance-based measures of motor or cognitive output. During the clinical management of fatigue, it is important to consider the role of other MS symptoms on fatigue, as well as that of non-MS related medical conditions. Management of fatigue in MS often entails both pharmacologic and behavioral components. This article reviews recent developments in the assessment, treatment, and pathogenesis of MS fatigue. PMID- 11898531 TI - Immunologic therapy for secondary and primary progressive multiple sclerosis. AB - Multiple sclerosis (MS) is generally considered an immune-mediated demyelinating disease, and treatments designed to modify the course of MS are immunosuppressive or immunomodulatory. Although most people with MS have a relapsing-remitting course initially, the majority will eventually experience a more gradual decline in neurologic function, termed secondary progressive MS. Some patients have gradual worsening from the beginning, termed primary progressive MS. Recent pathologic studies have revealed that axonal injury and neuronal degeneration are much more prominent in MS than previously recognized, and may be the explanation for the gradual decline in neurologic function that characterizes progressive MS. The results of several clinical trials in MS indicate that suppression of the immune-mediated inflammation may decrease the relapse rate in MS, but not stop the progressive loss of neurologic function. There are many promising approaches to this clinical dilemma, but none has been proven to be effective in stopping or retarding progressive MS. More well-designed, controlled, blinded, randomized clinical trials are needed to test these putative therapies. In the mean time, we should avoid subjecting patients to potentially dangerous and unproven regimens. PMID- 11898534 TI - Activated microglia: the silent executioner in neurodegenerative disease? PMID- 11898535 TI - Literature alert. PMID- 11898533 TI - Management of spasticity, pain, and paroxysmal phenomena in multiple sclerosis. AB - Multiple sclerosis (MS) is a disease with tremendous variability and innumerable symptoms. Among the more common symptoms is spasticity. Despite a lack of full knowledge of the physiology causing this phenomenon, successful treatments have been developed. Many of these have had a recent introduction. Pain and paroxysmal phenomena are surprisingly common in MS, but have not had the recognition their frequency deserves. It is not unusual to hear that they are rare in MS, but surprisingly they are all too common. Their management is changing as newer treatments are developed. PMID- 11898536 TI - Parkinson's disease: surgical options. AB - The introduction of levodopa revolutionized the treatment of Parkinson's disease. However, complications of therapy that diminish functional capacity eventually develop in the majority of patients. Studies in animal models have demonstrated that the parkinsonian state is associated with overactivity in the output nuclei of the basal ganglia. This provides a rationale for surgically targeting these nuclei to diminish this overactivity and reestablish a more balanced output (compensatory strategy). Lesioning and high-frequency stimulation of either the pallidum or the subthalamic nuclei are effective, but many questions remain regarding what surgery is best. Even more questions remain regarding the place of a restorative strategy, namely implantation of fetal midbrain tissue to replace the missing dopamine cells and "cure" the disease. Practical, ethical, and legal issues that complicate the use of human tissue have encouraged initial attempts at xenotransplantation using porcine fetal tissue. PMID- 11898537 TI - Parkinson's disease: the treatment of drug-induced hallucinations and psychosis. AB - Drug-induced psychosis is one of the most disabling complications of advancing Parkinson's disease. It has also been one of the most difficult to treat. Clozapine was the first medication shown to be safe and effective in this setting, and it remains the standard by which newer atypical antipsychotics are measured. However, due to the small but significant risk of agranulocytosis and the need for frequent blood testing, alternatives have been sought. Risperidone, olanzapine, and quetiapine are new atypical antipsychotics that have each been proposed as an alternative to clozapine, but the literature concerning their use in Parkinson's disease is conflicted and confusing. Although quetiapine appears to be the best current choice, none of these medications have equaled clozapine's ability to safely treat drug-induced psychosis without the risk of worsening parkinsonism. PMID- 11898538 TI - Initial treatment of early Parkinson's disease: a review of recent, randomized controlled trials. AB - Many studies have shown dopamine agonists to significantly improve parkinsonian symptoms compared with placebo in early Parkinson's disease (PD), but how do agonists compare with the standard treatment of levodopa? Recently, three large, multicenter, randomized controlled studies directly comparing a dopamine agonist with levodopa as initial therapy in early PD have been published. These studies suggest that although both agents effectively ameliorate parkinsonian symptoms, levodopa was superior to dopamine agonists as measured by improvement in Unified Parkinson's Disease Rating Scale (UPDRS) scores. However, levodopa was more frequently associated with dopaminergic motor complications, and the dopamine agonists were more commonly associated with adverse events. Until further studies clearly demonstrate the beneficial effects of one therapeutic strategy over another, the decision to initiate treatment in early PD with either an agonist or levodopa will be based on the favorable motor complication profile of agonists versus the more potent antiparkinsonian effects and the favorable side-effect profile of levodopa. PMID- 11898539 TI - Focal dystonia: the role of botulinum toxin. AB - Botulinum toxin (BTX) has been found to be effective in a wide range of focal dystonias. Debate surrounds the selection of injection sites. In general, localization is satisfactory by clinical examination, but poor response, requiring localization of deep muscles, may necessitate use of electromyography for localization. Delineation of optimal doses of BTX is a work in progress; as studies have tended to show efficacy at lower doses than used in the past, the trend is to use lower doses. This is important, because development of antibodies to BTX, the main reason for secondary resistance to this treatment, is more frequent with larger doses and shorter inter-injection intervals. Although the mechanism of denervation of the neuromuscular injunction by BTX is relatively well understood, secondary changes at the level of the basal ganglia, thalamus, and cortex, and their role in response to BTX, need further exploration. PMID- 11898540 TI - Measurement of quality of life in neurodegenerative disorders. AB - The importance of recognizing and documenting the subjective impact of disease and the side effects of long-term medical therapy in illnesses such as Parkinson's disease is beginning to be fully appreciated. This appreciation follows the recognition that it is necessary to describe the overall results of diagnostic and treatment efforts in a way that makes sense to both patients and health professionals. A recent transformation of thought and practice has occurred, where inclusion of concepts commonly subsumed under the term quality of life or health-related quality of life are considered for incorporation into clinical trials and clinical practice. Recent editorials illustrate this shift in emphasis [1]. PMID- 11898541 TI - Neurologic disorders of gait. AB - Gait disorders are important because of their prevalence, particularly among the elderly, and the associated risk of falls and injury. Neural networks that organize locomotion and maintain balance are briefly reviewed. Gait disorders can be classified based on observational features or by etiology. Several common disorders are discussed in more detail. Recent progress includes use of botulinum toxin for spastic gait in cerebral palsy, neurosurgical treatment of Parkinson's disease, and newer rehabilitation approaches to gait and balance training. PMID- 11898542 TI - Pregnancy and antiepileptic drugs. PMID- 11898543 TI - Advances in neuroimaging: non-substrate-directed partial epilepsy. AB - Contemporary neuroimaging studies using structural and functional techniques are critical in the evaluation of patients with localization-related epilepsy. Imaging procedures may be used to localize the epileptic brain tissue or determine the likely pathologic findings underlying the epileptogenic zone, or both. The diagnostic yield of magnetic resonance imaging (MRI) has been demonstrated in patients with partial epilepsy. The identification of an MRI epileptogenic lesion is almost invariably a reliable indicator of the site of seizure onset. Peri-ictal single photon emission computed tomography (SPECT) may be of particular benefit in patients with normal MRI studies. The use of neuroimaging in the care and management of patients with partial epilepsy is discussed here. PMID- 11898544 TI - New antiepileptic drugs. AB - Finding a niche for each antiepileptic drug has become challenging as more new agents are approved. In this article, the most recent information regarding these drugs is reported, with emphasis on the agents approved in the past 10 years. The evidence clearly shows that, despite limited Food and Drug Administration indications, all drugs are useful in monotherapy, and many have broader spectrums of action. Adverse effects can also help distinguish between agents. Familiarity with this latest information will help clinicians make the best choice for each of their patients with epilepsy. PMID- 11898545 TI - Epilepsy surgery: chance for a cure. AB - Epilepsy is a chronic disorder with medical, as well as psychosocial, consequences. Many patients with epilepsy are well controlled by medication and are able to tolerate the side effects of the antiepileptic drugs needed to control their disorder. However, there are many people who are either unable to gain seizure freedom with medications or are intolerant of the side effects of their drugs. In some of these patients, particularly those with mesial temporal sclerosis, surgery offers a viable treatment alternative and a chance for a cure rather than just lifelong management. PMID- 11898546 TI - Etiology, diagnosis, and treatment of nonepileptic seizures. AB - Psychogenic nonepileptic seizures (NES) can be classified into five categories. This review focuses on NES associated with emotional conflict, by far the most common and important group. Etiology is speculative, but the background histories of these patients are often similar. The presence of a trauma history, depression, post-traumatic stress symptoms, and the use of dissociation plus cognitive dysfunction possibly point to an organic etiology. The presentation of NES in children and adults is discussed, along with the differential diagnosis. The diagnostic differential is lengthy, with epileptic seizures of frontal lobe origin presenting a unique challenge. Diagnostic procedures are reviewed with an emphasis on the utility of hypnosis with seizure induction. Presenting the diagnosis to the patient, the role of the neurologist, and the role of the mental health consultant are reviewed. Issues in the doctor-patient relationship are also addressed, as well as the overall prognosis. PMID- 11898547 TI - Advances in our understanding of early childhood epilepsies: 1999-2000. AB - Epilepsy is frequent in early childhood, but poorly understood. Management of very young children with refractory epilepsy is particularly challenging, but recent advances in classification, medical therapy, and surgical treatment are helping. We summarize work in each area, focusing on developments in the past 2 years. Classification schemes unique to the needs of very young children will likely improve our ability to accurately diagnose and treat those with refractory seizures. Medication studies are now being done that address some of the specific epilepsy syndromes seen in infants and young children, with promising results in some circumstances. Progress is also being made in the identification of good candidates for early, effective epilepsy surgery. For a variety of reasons and incentives, it is likely that research focusing on infants and young children will continue at a brisk pace. PMID- 11898549 TI - Iatrogenic epidemic involuntary dementia: impending harvest of the cardiopulmonary resuscitation rage. PMID- 11898548 TI - Epilepsy in the elderly. AB - The active prevalence rate of epilepsy among persons over 65 years of age is approximately 1.5%, about twice the rate in younger adults. Treatment of epilepsy in the elderly is complicated by alterations in drug metabolism, use of concomitant medications, and multiple medical problems. Drug interactions are a major issue, and a full knowledge of the isoenzyme profile and protein-binding characteristics of each drug (antiepileptic and other) must be known. PMID- 11898550 TI - Literature alert. PMID- 11898551 TI - Frontotemporal dementia and tauopathy. AB - The presence of abundant neurofibrillary lesions made of hyperphosphorylated tau proteins is the characteristic neuropathology of a subset of neurodegenerative disorders classified as "tauopathies." The discovery of mutations in the tau gene in frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) constitutes convincing evidence that tau proteins play a key role in the pathogenesis of neurodegenerative disorders. Moreover, it now is known that the most common form of sporadic frontotemporal dementia (FTD), which is characterized by frontotemporal neuron loss, gliosis, and microvacuolar change, also is a tauopathy caused by a loss of tau protein expression. Thus, these discoveries have begun to change the classification and the neuropathologic diagnosis of FTD and tauopathies, as well as current understanding of the disease mechanisms underlying them. Although transgenic mice expressing wild-type human tau or variants thereof with an FTDP-17 mutation result in tau pathologies and brain degeneration similar to that seen in human tauopathies, the precise mechanisms leading to the onset and progression of neurodegenerative disorders remain incompletely understood. Here, we review current understanding of human neurodegenerative tauopathies and prospects for translative recent insights about these into therapeutic interventions to prevent or ameliorate them. PMID- 11898552 TI - Amyloid metabolism and secretases in Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by the progressive accumulation of amyloid fibrils composed of the amyloid beta-protein (A beta) in senile plaques. A beta is derived from the beta-amyloid precursor protein (APP) after beta- and gamma-secretase cleavages. beta-secretase was recently identified to be a membrane-anchored aspartyl protease that is widely distributed in subcellular compartments, including Golgi, trans-Golgi network, and endosomes. Although definitive identification of gamma-secretase will require reconstituting its activity in vitro, mounting evidence suggests that gamma-secretase is an unusual intramembrane-cleaving aspartyl protease. Two intramembranous aspartate residues in presenilin (PS) are absolutely required for A beta generation. Three classes of gamma-secretase inhibitors can directly bind to PS, strongly supporting the hypothesis of PSI as gamma-secretase. These results provide the molecular basis for therapeutic interventions that reduce A beta accumulation in AD patients by inhibiting beta- or gamma-secretase. PMID- 11898553 TI - Pharmacotheraphy for Alzheimer's disease. AB - In the past 2 years, substantive advances in therapy for Alzheimer's disease (AD) have occurred. The nature of the effects of cholinesterase inhibitors has been refined with the publication of several studies that have examined different aspects of the symptomatology of AD. Breakthroughs in the basic science of Alzheimer's disease have led to new insights into potential therapeutic strategies targeted at the secretases involved in the metabolism of the Alzheimer precursor protein. An immunization approach, in which the beta-amyloid protein itself was used as the immunizing agent, has also been presented and independently validated. Other areas of investigation with disappointing results, such as estrogen replacement therapy, anti-inflammatory approaches, and several other therapeutic agents, are also reviewed. PMID- 11898554 TI - Lewy bodies and dementia. AB - The discovery of widely distributed Lewy bodies (LBs) in the brains of patients with dementia has stimulated much clinical and pathologic inquiry. This clinico pathologic syndrome is now referred to as dementia with Lewy bodies (DLB). Diagnostic criteria for DLB proposed at a workshop in 1995 are receiving detailed scrutiny. The criteria are complex to apply, and appear to have high specificity, but variable sensitivity. Neuropathologic studies have been aided by the development of probes against alpha-synuclein, a key component of LBs. Widespread LBs in limbic or cortical areas contribute to dementia. Pharmacologic management of cognitive and behavioral symptoms in patients with DLB is being explored. There is evidence that cholinesterase inhibitors may have beneficial effects. PMID- 11898555 TI - Of replications and refutations: the status of Alzheimer's disease genetic research. AB - Alzheimer's disease (AD) is a genetically complex and heterogeneous disorder. To date, mutations in three genes (APP, PSEN1, PSEN2) have been described to cause familial early-onset AD. In addition, a common polymorphism in the gene encoding apolipoprotein E (APOE) has been associated with the more common late-onset form of the disease. However, many studies have shown that genetic factors other than APOE play an important role in late-onset AD. Along these lines, a recent report predicted the existence of at least four additional late-onset AD genes, one of which was estimated to have a much greater contribution to age of onset variation than the APOE epsilon 4-allele. However, most of the nearly three dozen loci that have been proposed as putative AD genes to date have been followed by both replications and refutations, making consensus impossible. In this overview, we discuss the current status of genetic research in AD, including a brief summary of applicable analytic tools, and a summary of recent findings suggesting the existence of novel AD genes on chromosomes 10, 11, and 12. PMID- 11898556 TI - New developments in animal models of Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by deterioration in mental function leading to dementia, deposition of amyloid plaques and neurofibrillary tangles (NFTs), and neuronal loss. The major component of plaques is the amyloid-beta peptide (A beta), whereas NFTs are assemblies of hyperphosphorylated forms of the microtubule-associated protein tau. Electron microscopy of NFTs reveals structures known as paired helical filaments (PHFs). In familial AD (FAD), mutations in three distinct genes drive A beta synthesis by favoring endoproteolytic secretase cleavages that liberate A beta from the Alzheimer beta amyloid precursor protein (APP). This suggests that excess A beta initiates a pathogenic cascade in humans that culminates in all the pathologic and cellular hallmarks of AD. Building upon the knowledge of FAD mutations, incremental technical advances have now allowed reproduceable creation of APP transgenic mice that exhibit AD-like amyloid pathology and A beta burdens. These transgenic mouse lines also exhibit deficits in spatial reference and working memory, with immunization against A beta abrogating both AD-associated phenotypes. Besides establishing a proof of principle for A beta-directed therapies, these findings suggest a potential to identify individual elements in the pathogenic pathway that lead to cognitive dysfunction. Furthermore, transgenic APP mice with potent amyloid deposition will likely form a beach-head to capture the final elements of AD neuropathology--cell loss and NFTs composed of PHFs--that are missing from current transgenic models. PMID- 11898557 TI - New developments in idiopathic intracranial hypertension. AB - This review highlights recent additions to the literature regarding the diagnosis, evaluation, and management of idiopathic intracranial hypertension (pseudotumor cerebri). Unique features of pediatric pseudotumor cerebri are addressed as well. PMID- 11898559 TI - Benign paroxysmal positional vertigo. AB - Benign paroxysmal positional vertigo (BPPV) is the most common and most treatable cause of vertigo. In most cases, a simple maneuver that takes less than a few minutes to do resolves the problem. BPPV is caused by misplaced calcium carbonate crystals (otoconia) in the semicircular canal of the inner ear that have broken free from the utricle. When these crystals break free, they either remain loose in one of three different semicircular canals or attach to the hair cells within a canal. Several different types of treatment maneuvers have been described. The maneuver to use varies according to the semicircular canal involved and whether the crystals are loose or attached to the hair cells. PMID- 11898558 TI - Nystagmus. AB - Advances in understanding the organization of the ocular motor system, including its anatomy and pharmacology, have provided new insights into the pathogenesis of various forms of nystagmus. The discoveries of fibromuscular pulleys that govern the pulling directions of the extraocular muscles has provided a new conceptual framework to account for the different axes of rotation of vestibular and other types of movements that may contribute to nystagmus. Theoretical and experimental evidence has suggested that acquired pendular nystagmus, which is commonly due to multiple sclerosis, arises from the neural network that normally guarantees steady gaze by integrating premotor signals. Pharmacologic inactivation studies have implicated both gamma-aminobutyric acid (GABA) and glutamate as important transmitters in the neural integrator and suggested new drug therapies. New electro-optic devices may eventually prove to be effective treatment for the visual symptoms cause by acquired nystagmus. The demonstration of proprioceptive mechanisms in the distal extraocular muscles has provided a rationale for new operative treatments for congenital nystagmus. PMID- 11898560 TI - Perilymphatic fistula. AB - A perilymph fistula is an abnormal connection between the inner and middle ear that allows escape of perilymph fluid into the middle ear compartment. The clinical symptomatology that follows leakage of fluid is by and large indistinguishable from a number of other pathologies that affect inner ear function. Definite diagnostic proof of a perilymph fistula remains elusive, and methods of diagnosis remain controversial. Traumatic tears in the oval or round windows remain a major cause of perilymph fistula, yet an index of suspicion in traumatic brain injury frequently remains low. The diagnosis of perilymph fistula must always be considered in the appropriate clinical setting of head trauma, barotrauma, and in patients with unresolved and undiagnosed episodes of recurrent vertigo or hearing loss. Surgical treatment with patching of oval and round windows remains the mainstay of therapy for this condition. PMID- 11898562 TI - Evidence basis for treatment of spasticity. PMID- 11898563 TI - Literature alert. PMID- 11898561 TI - Tinnitus. AB - Tinnitus is a common otologic symptom secondary to numerous etiologies, such as noise exposure, otitis, Meniere's disease, otosclerosis, trauma, medications, and presbycusis. A thorough evaluation is necessary to rule out less common causes, which may include acoustic neuromas, glomus tumors, atherosclerosis of the carotid arteries, arteriovenous fistulae (AVFs), arteriovenous fistulae malformations (AVMs), and intracranial hypertension. Treating physicians need to have a very compassionate attitude towards these patients, and statements such as "there is nothing that can be done" are very inappropriate and should be strongly condemned. Reassurance, hearing aids, masking devices, retraining methods, antidepressants, intratympanic medications, and management of underlying pathologies such as carotid artery atherosclerosis, skull base tumors, intracranial hypertension, and AVMs/AVFs provide relief for the majority of these patients. PMID- 11898564 TI - Clinical applications of new cerebrospinal fluid analytic techniques for the diagnosis and treatment of central nervous system infections. AB - Advances in molecular biology and immunology provide new, highly sensitive and specific techniques that can be applied to analysis of cerebrospinal fluid to enhance the diagnosis and treatment of central nervous system (CNS) infections. In addition to improved accuracy and speed of diagnosis, these modalities may offer improved means of monitoring treatment efficacy, establishing prognosis, detecting organism resistance, and tracking epidemic sources. This brief review discusses a number of recent papers applying these methods, in order to illustrate their value and significance for clinical neurologic practice. Some of these applications are commonly available, whereas others are likely to enter the physician's armamentarium in the near future. As they do, they can be expected to improve the treatment of CNS infections. PMID- 11898565 TI - Herpes zoster infection and postherpetic neuralgia. AB - Varicella-zoster virus (VZV), the cause of chicken pox, establishes latent infection in sensory ganglia. Reactivation results in zoster (shingles), sometimes complicated by encephalitis (myelitis). Postherpetic neuralgia (PHN) is the major morbidity of zoster. PHN typically increases in frequency with age. The VZV vaccine, which was developed for children, may be effective in enhancing VZV immune reactivity and decreasing zoster in adults. Early antiviral treatment may be effective in decreasing PHN onset. Several other medications may be useful in treating established PHN. A recent report discussed intrathecal steroid use. PMID- 11898566 TI - AIDS and AIDS-treatment neuropathies. AB - AIDS and AIDS-treatment neuropathies are common in individuals infected with HIV. As patients live longer due to improved antiretroviral therapies, the impact of painful neuropathy on patients' lives may increase. Several antiretroviral medications are known to cause toxic neuropathy in AIDS patients; but this may be outweighed by the beneficial effects of viral suppression. Current theories on the pathogenesis of AIDS neuropathies include mitochondrial toxicity secondary to gamma-DNA polymerase inhibition and subsequent abnormal mitochondrial DNA synthesis. Treatment of AIDS neuropathies is directed toward relief of symptoms, however, new evidence suggests that aggressive antiretroviral therapy may also be effective. PMID- 11898567 TI - Spatial neglect. AB - Neglect, a failure to attend or act in one part of space, is a common disturbance following cerebral lesions. This article outlines the signs and symptoms of neglect. Recent attempts to disentangle sensory/attentional from motor/intentional aspects of neglect are outlined. Neglect in directions other than the horizontal is discussed, as are directional biases found in normal control subjects. The article also examines the anatomy of lesions causing neglect and theoretical models of the cerebral mechanisms underlying neglect. PMID- 11898568 TI - Frontal lobe functions. AB - The frontal lobes constitute two thirds of the human brain, yet the functions performed by them remained mysterious for a long time. Apart from their well known involvement in motor function and language, little was previously known about the functions of the frontal lobes. Recent advances have uncovered important roles for the frontal lobes in a multitude of cognitive processes, such as executive function, attention, memory, and language. The importance of the frontal lobes in processes underlying affect, mood, personality, self-awareness, as well as social and moral reasoning, is also a renewed area for research. This article focuses on recent advances in understanding frontal lobe functions. PMID- 11898570 TI - Prophylactic hyperdynamic postoperative fluid therapy after aneurysmal subarachnoid hemorrhage. PMID- 11898569 TI - Neuroimaging and behavior: probing brain behavior relationships in the 21st century. AB - Functional neuroimaging over the past decade has provided a new way to examine brain behavior relationships. Current noninvasive neuroimaging techniques, which can examine structure and function, have begun to clarify the networks involved in cognitive processes and how these are affected in aging and disease. Functional magnetic resonance imaging (fMRI) has demonstrated the interaction between medial temporal and prefrontal regions in episodic memory. The anatomical correlates of various components of spatial attention and working memory have emerged from elegant event-related fMRI designs. Distinct neural networks for different emotions are being mapped out, and the role of the anterior cingulate in depressed mood has been documented. This review highlights key recent studies that have illuminated the neural substrates of these important cognitive and affective processes. PMID- 11898571 TI - Critical care of intracerebral and subarachnoid hemorrhage. AB - The acute management of primary intracerebral or aneurysmal subarachnoid hemorrhage requires a comprehensive approach involving stabilization of the patient, surgical intervention, and continued intensive care treatment of medical and neurologic complications. The are several causes of intracerebral hemorrhage (ICH), including hypertension, cerebral amyloid angiopathy, sympathomimetic drugs, and coagulopathies. More recently, use of thrombolytic agents in the treatment of acute ischemic stroke has increased the risk of ICH. Treatment of intracerebral hemorrhage is based on blood pressure control, and, in selected cases, surgical evacuation of clot. Patients with aneurysmal subarachnoid hemorrhage may experience rebleeding, symptomatic vasospasm, or hydrocephalus. Medical management in the intensive care unit with careful attention to fluid and electrolyte balance, nutrition, cardiopulmonary monitoring, and close observation for changes in the neurologic exam is vital. This review examines the diagnosis and intensive care management of patients with intracerebral or subarachnoid hemorrhage, and reviews some of the newer therapies for treatment of these disorders. PMID- 11898572 TI - Central nervous system infections: a critical care approach. AB - Central nervous system (CNS) infections are diverse. CNS infections can cause significant morbidity and mortality and are markedly different from systemic infections. The closed anatomic space of the CNS, its immunologic isolation from the rest of the body, and the often nonspecific nature of the key manifestations present a challenge to the clinician. Early recognition and aggressive management are essential to patient recovery and prevention of long-term neurologic sequelae. This review discusses the major types of CNS infections and focuses on critical care management, with emphasis on current epidemiologic trends. PMID- 11898573 TI - Critical care of acute ischemic stroke. AB - Advances in neurologic therapeutics and intensive care medicine have expanded the arsenal of treatments available for the critical care of ischemic stroke. Several agents are available for acute reperfusion of the ischemic brain. These include intravenous recombinant tissue-plasminogen activator (rtPA), which is effective in a 3-hour time window, and intra-arterial thrombolytics, which may be effective within 6 hours. In addition, newer agents such as Ancrod and abciximab may be effective within the acute time period. Efforts to prevent secondary brain injury in critically ill patients with stroke often include prevention and reduction of fever, induced hypertension, and mechanical ventilation. Finally, death due to severe brain edema after massive hemispheric infarction can often be prevented with surgical or medical intervention. Unfortunately, there is a critical lack of well-designed clinical studies to guide the clinician in the use of these interventions. In addition, there is concern that some of these interventions may preserve life at the cost of quality of life. This article reviews the evidence behind these approaches to the critical care of ischemic stroke. PMID- 11898574 TI - Outcomes of neurocritical care. AB - Outcomes research is a new and developing field that attempts to better measure the results of medical care. Neurocritical care is a relatively new field of medicine focusing on the care of critically ill patients with primary or secondary neurologic problems. Much needs to be done to examine the outcomes of neurocritical care, and the following is a review of the pertinent concepts of both outcomes and neurocritical care research. PMID- 11898575 TI - Is the promise of randomized control trials ("evidence-based medicine") overstated? PMID- 11898577 TI - Angioplasty and stenting for severe stenosis of the extracranial portion of the internal carotid artery. PMID- 11898578 TI - Diagnosis and treatment of ischemic pediatric stroke. AB - Tremendous progress has been made in the diagnosis, prevention, and treatment of pediatric stroke. With a complete investigation, stroke etiology can be determined in most children, and multiple factors are commonly identified. Stroke can be prevented in some children and treated in others. Children at risk for recurrent strokes can be treated effectively. The prognosis after pediatric stroke is usually good, but today we can identify and initiate treatment in selected patients at risk for long-term problems. This article reviews recent advances in the identification, prevention, treatment, and outcome in pediatric ischemic stroke. PMID- 11898580 TI - Selection of anticoagulants or antiplatelet-aggregating agents for prevention of stroke. AB - Stroke is one of leading causes of mortality and morbidity in the United States. Stroke prevention includes treatment of the stroke risk factors and long-term use of antithrombotic agents. Various agents have been studied for stroke prevention and other trials are ongoing. The aim of this article is to provide an overview of the recent guidelines, recommendations, and clinical trial results using antithrombotic therapy for stroke prevention. PMID- 11898579 TI - Utility of noninvasive studies in the evaluation of patients with carotid artery disease. AB - Endarterectomy reduces the risk of stroke in selected patients with carotid artery stenosis, and the benefit is related to the degree of stenosis. Although the randomized trials demonstrating this benefit measured the degree of stenosis with conventional catheter angiography, many physicians are relying on noninvasive tests to select patients for surgery. Technologic advancement in this area is outpacing the availability of quality data supporting the clinical utility of the newer noninvasive tests. PMID- 11898582 TI - Treatment of arteriovenous malformations of the brain. AB - Arteriovenous malformations (AVMs) of the brain are complex, congenital, high flow vascular lesions in which there is a direct fistulous connection between arteries and veins without an intervening capillary bed. Patients commonly present with hemorrhage or seizures. Treatment must take into account the natural history of these lesions and the risks related to their treatment. To date, there have been no prospective, randomized, controlled clinical trials to determine the optimal method of treatment of brain AVMs. However, multiple retrospective analyses of treatment strategies and surgical results in patients with brain AVMs reveal that the management of brain AVMs requires a multimodal, multidisciplinary treatment approach, utilizing surgical resection, endovascular embolization, and/or stereotactic focused-beam radiosurgery. PMID- 11898583 TI - Aminoglycoside treatment for muscular dystrophy is scientifically rational, but is it clinically effective? PMID- 11898581 TI - Hemorrhagic transformation following ischemic stroke: significance, causes, and relationship to therapy and treatment. AB - Hemorrhagic transformation (HT) is a frequent consequence of ischemic stroke that becomes more prevalent after thrombolytic therapy. Despite concerns about safety parameters, thrombolytic drugs remain the first course of action available to clinicians for stroke management. However, recent efforts in preclinical studies have attempted to discover other drugs that can lessen the risk of hemorrhage associated with thrombolytic administration. This review focuses on three classes of pharmacologic agents that have shown some promise in animal models of stroke, and can thus be considered as possible candidates for coadministration with thrombolytics in the treatment of stroke. These include the following: 1) spin trap agents, such as alpha-phenyl-N-t-butylnitrone (PBN) that scavenge free radicals; 2) matrix metalloproteinase (MMP) inhibitors, such as BB-94, that prevent membrane and vessel remodeling following ischemia; and 3) the novel glycoprotein (GP) IIb/IIIa platelet receptor antagonist SM-20302. Although these drugs affect different mechanisms, the common denominator seemed to be their effectiveness in reducing the incidence of hemorrhage in response to thrombolytic infusion following an embolic stroke. PMID- 11898584 TI - Amyotrophic lateral sclerosis: objective upper motor neuron markers. AB - The diagnosis of amyotrophic lateral sclerosis (ALS) remains a clinical diagnosis. It is based on the combination of both upper and lower motor neuron signs in the neurologic examination. With several new therapeutic agents on the horizon, effective and objective disease markers for diagnosis and surrogate outcome measures in clinical trials are crucial. Whereas the presence of lower motor neuron signs on neurologic examination can be ascertained by electromyography, there is no widely accepted marker for upper motor neuron involvement. Neuroimaging changes of the corticospinal tract in ALS patients have been studied using magnetic resonance (MR) imaging, but appear to lack sensitivity and specificity. MR spectroscopy, a technique that allows one to evaluate biochemical tissue composition in vivo, has been widely used to establish the progressive decrease in N-acetylaspartate, a marker of neuronal integrity, in the course of ALS. More recently, diffusion tensor imaging, a newer MR technique, has demonstrated changes in diffusivity along the corticospinal tract in ALS patients. Metabolic aspects in the brains of ALS patients have been evaluated using positron emission tomography. Transcranial magnetic stimulation is a more established technique that evaluates the neurophysiologic integrity of upper motor neurons in ALS. This article reviews the progress that has been made over the past two decades towards establishing valid diagnostic and natural history markers of upper motor neuron involvement in ALS. PMID- 11898585 TI - Periodic paralysis: understanding channelopathies. AB - Familial periodic paralyses are typical channelopathies (i.e., caused by functional disturbances of ion channel proteins). The episodes of flaccid muscle weakness observed in these disorders are due to underexcitability of sarcolemma leading to a silent electromyogram and the lack of action potentials even upon electrical stimulation. Interictally, ion channel malfunction is well compensated, so that special exogenous or endogenous triggers are required to produce symptoms in the patients. An especially obvious trigger is the level of serum potassium (K+), the ion responsible for resting membrane potential and degree of excitability. The clinical symptoms can be caused by mutations in genes coding for ion channels that mediate different functions for maintaining the resting potential or propagating the action potential, the basis of excitability. The phenotype is determined by the type of functional defect brought about by the mutations, rather than the channel effected, because the contrary phenotypes hyperkalemic periodic paralysis (HyperPP) and hypokalemic periodic paralysis (HypoPP) may be caused by point mutations in the same gene. Still, the common mechanism for inexcitability in all known episodic-weakness phenotypes is a long lasting depolarization that inactivates sodium ion (Na+) channels, initiating the action potential. PMID- 11898586 TI - The genetic convergence of Charcot-Marie-Tooth disease types 1 and 2 and the role of genetics in sporadic neuropathy. AB - Charcot-Marie-Tooth (CMT) disease represents a clinically and genetically heterogeneous group of inherited neuropathies caused by aberration of the intimate relationship between the myelin sheath and the axon; disorders causing demyelination are classified as CMT1 and those causing axonal loss as CMT2. The mechanisms by which mutations disturb the relationship of the myelin sheath and axon are not fully understood; however, we hypothesize that some mutations affect this relationship more profoundly than others, and thus account for the paradox that mutation of a "myelin gene" can present with electrophysiologic features of CMT2 and vice versa. Also, contrary to popular understanding, inherited neuropathies account for a substantial number of chronic peripheral neuropathies. Because of this observation, we propose that molecular diagnosis is a necessary adjunct for differentiating genetic and acquired peripheral neuropathies, even in sporadic chronic neuropathy. PMID- 11898587 TI - Congenital myasthenic syndromes: genetic defects of the neuromuscular junction. AB - Congenital myasthenic syndromes (CMS) stem from defects in presynaptic, synaptic, and postsynaptic proteins. The presynaptic CMS are associated with defects that curtail the evoked release of acetylcholine (ACh) quanta or the resynthesis of ACh. Insufficient resynthesis of ACh is now known to be caused by mutations that reduce the expression, catalytic efficiency, or both of choline acetyltransferase. The synaptic CMS are caused by mutations in the collagenic tail subunit (ColQ) of the endplate species of acetylcholinesterase that prevent ColQ from associating with catalytic subunits or from insertion into the synaptic basal lamina. With one exception, postsynaptic CMS identified to date are associated with a kinetic abnormality or decreased expression of the acetylcholine receptor (AChR). Numerous mutations have now been identified in subunits of AChR that alter the kinetics or surface expression of the receptor. The kinetic mutations increase or decrease the synaptic response to ACh and result in slow- and fast-channel syndromes, respectively. Most mutations that reduce surface expression of AChR reside in the receptor's epsilon subunit and are partially compensated by residual expression of the fetal-type gamma subunit. Null mutations in both alleles of other AChR subunits are likely lethal, owing to absence of a substituting subunit. PMID- 11898589 TI - Healing touch certificate program. PMID- 11898588 TI - Advances in myasthenia gravis. AB - Recent advances in the diagnosis and treatment of acquired myasthenia gravis (MG) are reviewed. Increased awareness about the need for more uniform methods of reporting treatment trials for MG has prompted systematic review of the literature and inspired an effort to develop better classifications and disease specific outcome measures. New antibodies have been discovered in patients with seronegative MG, possibly defining an immunologically distinct form of the disease. A new immunosuppressant, mycophenolate mofetil, may be an additional and safe option in the treatment of MG. Other work supports the possibility of developing a vaccine against MG suitable for trial in humans. PMID- 11898591 TI - Chi-Gung healing. PMID- 11898590 TI - The yoga path. PMID- 11898592 TI - [Na+/H+-exchanger in tissues of vertebrates]. PMID- 11898593 TI - [Ammonium compounds with localized and nonlocalized charges as reversible inhibitors of choline esterases of different origin]. PMID- 11898594 TI - [Activity of Na+, K+-Atpase and membrane lipids of the brain cells of trouts and rats under atmospheric pressure of 101]. PMID- 11898595 TI - [Registration of oxidation process of hemoglobin and cryoprotective effect of glycerine]. PMID- 11898596 TI - [The methemoglobin level in blood and resistant circulating erythrocytes of Scorpaena porcus in osmotic shock under experimental hypoxia conditions]. PMID- 11898597 TI - [Electrophysiological study of space distribution of vestibulospinal neurons of the frog Rana ridibunda]. PMID- 11898598 TI - [Interaction of tonic and phase type of pain in rabbit ontogenesis]. PMID- 11898599 TI - [Movement representation of facial muscles and vibration in the brain of white mice Mus musculus]. PMID- 11898600 TI - [Oxygen requirement for Baikal Pusa sibirica and factors, influencing it]. PMID- 11898601 TI - [Comparative physiological study of the specifics of recognition of difficult visual images and ability to identify two-dimensional pictures and three dimensional objects by high and low monkeys]. PMID- 11898602 TI - [Morphologic and functional characteristics of gonadotropin-releasing hormone immune reactive structures in the brain of bony fishes Acipenser guldenstadti before reproduction behavior]. PMID- 11898603 TI - [Increase in brain activity of the carp Cyprinus carpio during visual deafferentation]. PMID- 11898604 TI - [Neurons in the inferior colliculus of the mouse Mus musculus are sensitive to the spectral changes in noise]. PMID- 11898605 TI - [Structural changes in the sensomotor part of the brain in early postnatal ontogenesis of rats, who suffered prenatal hypoxia]. PMID- 11898606 TI - [Comparative genomic hybridization as a new method for detection of genomic imbalance]. AB - Comparative Genomic Hybridization (CGH) is a molecular cytogenetic analysis that allows identification of genomic changes by comparing the copy number of DNA sequences in cells of tested tissue and the reference specimen. CGH is based on competitive suppressive in situ hybridization of two differently labeled DNA probes (tested and reference, karyotypically normal, fluorochrome-labeled DNAs) with metaphase chromosomes of a healthy subject. First described by Kallioniemi et al. in 1992, the CGH assay has been widely used for identification and characterization of both numerical and structural chromosome abnormalities in cells of different tissues at various pathological conditions in humans, especially in tumor diseases. We discuss the specific features and quality control of comparative genomic hybridization, its advantages and limitations in detection of genomic imbalance and the prospects for development of this technology. PMID- 11898607 TI - [The mitochondrial genome and human mitochondrial diseases]. AB - To date, more than 100 point mutations and several hundreds of structural rearrangements of mitochondrial DNA (mtDNA) are known too be connected with characteristic neuromuscular and other mitochondrial syndromes varying form those causing death at the neonatal stage to diseases with late ages of onset. The immediate cause of mitochondrial disorders is a defective oxidative phosphorylation. Wide phenotypic variation and the heteroplasmy phenomenon, which some authors include in mutation load, are characteristic of human mitochondrial diseases. As the numbers of cases identified and pedigrees described increase, data on the genotype--phenotype interaction and the structure and frequency of pathogenic and conditionally pathogenic mtDNA mutations in human populations are rapidly accumulated. The data on the genetics and epidemiology of mitochondrial diseases are not only important for differential diagnosis and genetic counseling. Since both neutral and mildly pathogenic mutations of mtDNA are progressively accumulated in maternal phyletic lines, molecular analysis of these mutations permits not only reconstruction of the genealogical tree of modern humans, but also estimation of the role that these mutations play in natural selection. PMID- 11898608 TI - [Study of RecA-independent homologous recombination and a chromosomal rearrangement in the Escherichia coli strain carrying an extended tandem duplication]. AB - A heterozygous tandem duplication in the Escherichia coli deo operon region deoA deoB::Tn5/deoC deoD thr::Tn9 with the total length approximately 150 kb, which was obtained in the conjugational mating in the HfrH strain, was examined. By means of digestion with the NotI enzyme, pulsed-field gel electrophoresis, and the conjugational transfer of the duplication in the F- strain, the chromosomal rearrangement, which occurred in the duplication region upon its stabilization in the bacterial genome, was studied. In a more stable strain, two new NotI sites were shown to appear in the chromosomal region located close to the duplication, which might have resulted from the transposition of the IS50 sequence from Tn5. The data were also obtained indicating the possibility of secondary transposition of the chromosomal segment between the two new NotI sites (approximately 30 kb) in the region located near the duplication. With the use of rec+ and recA strains, two types of haploid and diploid segregants generated by the duplication were studied: DeoD+ (the DeoD+ allele is not expressed in the original duplication due to the polar effect of the deoB::Tn5 insertion) and DeoC DeoD. The segregation of DeoD+ clones was shown to be RecA-dependent, whereas the DeoC DeoD segregants selected on the medium that contained thymine at a low concentration (i.e., under conditions of thymine starvation) appeared at a rather high frequency. However, the relative frequency of haploid clones, which have lost the duplication, strongly decreased in the recA genome among segregants of both types. PMID- 11898609 TI - [The use of fragments of the 85- and 120-MDa plasmids of Azospirillum brasilense Sp245 to study the plasmid rearrangement in this bacterium and to search for homologous sequences in plasmids of Azospirillum brasilense Sp7]. AB - A spontaneous loss of the 85- (p85) and 120-MDa (p120) replicons and simultaneous generation of a plasmid of more than 300 MDa were associated with defects in synthesis of O-specific and Calcofluor-binding polysaccharides and had no effect on flagellation and motility of the Azospirillum brasilense Sp245.5 mutant. The plasmid rearrangement was studied by hybridization of DNAs from the wild-type Sp245 strain and the Sp245.5 mutant with p85 and p120 fragments that contained loci involved in formation of the polar (fla) and lateral (laf) flagella, synthesis of O-specific and Calcofluor-binding polysaccharides (lps/cal), swimming (mot), and swarming (swa) of bacteria. Hybridization with the p120 fragments revealed incorporation of the intact fla/swa loci and the altered lps/cal loci into a new megaplasmid. Two EcoRI fragments homologous to the fla/laf/mot/swa loci of p85 were found in A. brasilense Sp245 DNA, whereas only one copy was preserved in the Sp245.5 mutant. Hybridization of the p120 and p85 fragments of Sp245 to the A. brasilense Sp7 DNA for the first time revealed regions of substantial homology to these fragments in the 90- and 115-MDa Sp7 plasmids, respectively. PMID- 11898610 TI - [Analysis for pheromone-induced cytogenetic disturbances depending on major urinary proteins of male laboratory mice]. AB - Young male CBA/LacStoRap mice for 2 h were exposed to pheromones that are transferred by major urinary proteins of sexually mature males of the same line. The treatment was conducted using either a complete set of the major urinary proteins typical of the CBA mice or incomplete set of protein fractions detected in some animals. The effect of pheromones was estimated 24 h after treatment by cytogenetic analysis for disturbances in dividing bone marrow cells at anaphase- telophase and in germ cells at metaphase I. The frequency of both mitotic and meiotic disturbances was significantly increased after exposure to pheromones associated with the complete spectrum of the major urinary proteins. Conversely, no cytogenetic effect was observed in the absence of particular protein fractions. Possible consequences of the pheromonal effect on the genomes of somatic and germ cells are discussed as well as the relationships between pheromones and the major urinary proteins. PMID- 11898611 TI - [The role of genotype in the formation of Leydig cell hormone function during postnatal ontogeny in diallele crosses of laboratory mice]. AB - Production of testosterone by Leydig cells during the postnatal ontogeny in pubescence under in vitro stimulation by chorionic gonadotropin, dibutiryl-cAMP, and pregnenolon was studied in males of four inbred mouse lines (BALB/c, RT, CBA/Lac, and A/He) and their F1 reciprocal hybrids. Highly statistically significant association between the animal genotype and age was revealed for all parameters studied, which indicates the genotype-dependent formation of the Leydig cell hormone function during the postnatal ontogeny. The effect of genotype was characterized by two specific features. First, in each postnatal ontogeny stages examined correlative genetic variability in respect of the cAMP- and substrate-dependent indices of Leydig cell reactivity was observed. Second, during postnatal ontogeny coordinated genetic variability was subjected to substantial ontogenetic rearrangements. Definite pattern of genetic differences in the Leydig cell hormone activity was formed only at the late pubertal--early post- pubertal stage (60th day after birth). This process coincided with the completion of the Leydig cell morphological differentiation and the appearance of mature cells in the population. Thus, formation of the Leydig cell hormone activity during postnatal ontogeny is under coordinated genetic control, which is also subjected to substantial changes during pubertal differentiation. PMID- 11898612 TI - [Genetic differentiation of pedunculate oak Quercus robur L. in the European part of Russia based on RAPD markers]. AB - Using randomly amplified polymorphic DNA markers (RAPD), genetic variation and differentiation in four populations of pedunculate oak Quercus robur L. were examined. The populations occupy a large part of the Quercus robur range in the European Russia (Voronezh and Novgorod oblasts; Republics of Mordovia and Bashkortostan). With each of six random primers (A02, A09, A17, B01, B08, B11), 96 DNA samples were analyzed by PCR. In all, 48 putative polymorphic RAPD loci were detected. We failed to reveal population-specific DNA fragments for any primer although the frequencies of 14 fragments were significantly different among populations. The oak populations studied exhibited high variability: 73-90% of genes were polymorphic and the effective allele number was about 1.4. The total genetic variation varied from 0.202 (Vor) to 0.245 (Nov), which corresponded to the estimates for populations of this species from Central and Western Europe. The populations examined showed low among-population differentiation (GST = 0.098); gene flow Nem was 4.61. The proportion of among population variation of the RAPD loci studied accounted for 7% of the total variability; more than 93% of the total variability was explained by individual and within--population variation. PMID- 11898613 TI - [The expression of mutation sy2 causing nonhomologous synapsis in meiosis of diploid rye Secale cereale L]. AB - The cytological expression of spontaneous mutation sy2 isolated from a population of weedy rye was examined. It was demonstrated that the primary defect of meiosis in the mutant plants is nonhomologous synapsis, which occurs simultaneously with the homologous one. An electron microscope study of the synaptonemal complex (SC) at prophase I showed synaptic abnormalities that manifested as "switches" of synapting axial elements to the nonhomologous partner and the formation of foldbacks of lateral SC elements. The sy2 mutants are characterized by one to two such events per meiosis. Nonhomologous synapsis leads to the appearance of univalents at metaphase I (on average 4.16 +/- 0.022 per meiocyte) and multivalents (on average 0.12 +/- 0.007 per meiocyte). The presence of multivalents in 12.0% of meiocytes at metaphase I may result from recombination in ectopic regions of homology. It is suggested that the sy2 mutation impairs a component of the system that limits synapsis in meiocytes to only homologous chromosome pairs. PMID- 11898614 TI - [Statistical analysis of secondary sex ratio variation in sable Martes zibellina L]. AB - The secondary sex ratio in sable Martes zibellina L. maintained in captivity was estimated for the first time ever. The data obtained at the Pushkin pedigree breeding farm (Moscow oblast) in 1982 through 1987 were analyzed. In total, 1705 litters of 414 females were examined. The total frequency of male births (P) was 0.527 +/- 0.007; the 95%-confidence interval of p (the probability of birth of a male) was within the limits 0.513 << p << 0.541, and the deviation from the expected 1:1 ratio was statistically significant. No effect of parental age and litter size on the number of male progeny was found. This may indicate a small influence of the parental hormonal and immunological status on sex ratio, which was reported in many other mammal species including those related to sable. Apparently, there is an evolutionary mechanism underlying the stable excess of males in sable litters. PMID- 11898615 TI - [Population genetic study of the Alatyr region of the Republic of Chuvashiia]. AB - Population genetic characteristics were estimated in the Alatyr' raion (administrative district) of the Republic of Chuvashia, which has long been populated by three ethnic groups. The ethnic assortativeness values in the town of Alatyr' and the rural area of the district were 1.17 and 1.21, respectively, for Russians; 1.14 and 4.82, respectively, for Chuvashes; and 1.33 and 2.45, respectively, for Mordovians. Wright's statistics were as follows: Fst = 0.00358, Fit = 0.00178, and Fis = 0.00134. The migration indices were 0.0264 for Alatyr' and 0.0178 for the district. The endogamy indices for the total and the Russian populations of Alatyr' were 0.47 and 0.53, respectively. The parameters of isolation by distance were a = 0.000189 and b = 0.00959 for the urban and a = 0.000318 and b = 0.00919 for the rural area. Schemes of the genetic landscape were constructed. The influence of the polyethnic composition on the genetic structure of the population is discussed. PMID- 11898616 TI - [Analysis of polymorphism of the number of tandem repeats in the aggrecan gene exon G3 in the families with idiopathic scoliosis]. AB - In our previous study we showed that the inheritance of pronounced forms of idiopathic scoliosis was described by an autosomal-dominant major gene model assuming incomplete sex- and age-dependent penetrance. In the present study a search for the major gene was carried out by means of testing candidate genes. The aggrecan gene with known polymorphism of the number of tandem repeats in exon G3 was considered to be one of these candidate genes. Various alleles of this gene provide attachment of different number of chondroitin sulfate chains to a proteoglycan core protein, thereby changing functional properties of cartilage. Using the TDT analysis of 33 unrelated families consisting of a proband and his parents, we examined the existence of associations between the aggrecan alleles and the disease. Among nine alleles identified, three alleles with tandem repeats numbers of 25, 26, and 27 prevailed. We did not reveal preferable transmission of any of these alleles to the proband (TDT-statistics for different alleles varied from 0 to 0.71). There was also no correlation between the number of tandem repeats and the disease severity. Thus, either the polymorphism of the number of tandem repeats is not the direct reason for development of idiopathic scoliosis in the families tested, or its effect is too low to be detected using the samples examined. PMID- 11898618 TI - [Polymorphism of trinucleotide repeats at loci FRAXA and FRAXE in the population of Tomsk]. AB - Polymorphism of CGG and GCC trinucleotide repeats, whose expansions at the FRAXA and FRAXE loci have been identified as causative mutations in two forms of mental retardation, was studied in Slavic population of Tomsk. At the FRAXA locus a total of 31 allelic variants ranging from 8 to 56 copies of CGG repeat with two modal classes of 28-29 and 18-20 repeat units (with the frequencies of 24.6 and 11.5% respectively) were revealed. Compared to other populations, this locus was characterized by unusually high frequency of intermediate alleles with the sizes of more than 40 CGG repeat units (12.4%). Since intermediate repeats of the FRAXA locus were more prone to instability than normal alleles, it was suggested that Slavic population of Siberia had higher risk of the development of FMR1 dynamic mutations, giving rise to the Martin-Bell syndrome. The FRAXE allele frequency distribution was demonstrated to be normal with 18 allelic variants ranging from 9 to 27 GCC repeat units. In the population of Tomsk this locus had higher than in other populations frequency (26.7%) of short (less than 15 repeat units in size) alleles. In addition, in the Tomsk population both loci were characterized by high level of heterozygosity and low frequencies of modal allele classes. These results can be explained by the high level of outbreeding typical of the population of Siberia. PMID- 11898617 TI - [Micronuclei and ageing in a sample of Yugoslavian population]. AB - OBJECTIVE: Instability in the organization and expression of the genetic material has been hypothesized as the basic mechanism of ageing. Aim of this study was to quantify the effect of ageing on spontaneous micronuclei (MN) frequency in peripheral blood lymphocytes. METHOD: Analysis of Yugoslavian population sample (164 tested individuals, age 0-62 years) has performed by application of cytokinesis-block technique (CB). RESULTS: There was an increase of MN frequency with age, from newborns to 40-year-old persons. The total average of MN frequency per 1000 analyzed binuclear cells amounts to 8.03 +/- 0.42, with variation from 0 to 26 MNs. In a sample of 29 newborns the recorded average MN frequency was 6.91 +/- 0.81, while among 69 persons 25-39 years old, the MN frequency was 9.16 +/- 1.00. The lowest average MN frequency, however, was noted in the sample of 34 tested individuals 40 to 62 years of age. CONCLUSION: An increase with age in MN frequency has been observed in samples of studied individuals from newborns to 40 year-old persons. A decrease of MN frequency in older groups could be explained by a gradual decrease of proliferative cell capacities. PMID- 11898619 TI - [Transgenic tobacco (Nicotiana tabacum SR1) plants expressing the gene coding for Serratia marcescens nuclease]. AB - The gene coding for the secreted Serratia marcescens endonuclease was fused with the mannopine synthase promoter of Agrobacterium tumefaciens Ti plasmid and transferred to Nicotiana tabacum SR1 plants. The promoter is leaf- and root specific. The resulting transgenic plants demonstrated elevated nuclease activity. The level of the transgene product was determined in the transgenic lines. PMID- 11898620 TI - [Frequencies of the CCR2-64I and SDF1-3'A alleles associated with progression of the HIV-1 disease in healthy individuals from Moscow]. AB - The frequencies of two mutations associated with the development of clinical symptoms upon infection with human immunodeficiency virus type 1 (HIV-1) were determined in a cohort of individuals from Moscow. Allelic frequency of the first mutation, CCR2-64I, causing the substitution of valine with isoleucine in the CCR2 chemokine receptor, was 0.1106 (95% confidence interval, 0.0714-0.1498). The frequency of the second mutation the G to A substitution in the 3'-untranslated region of the stromal-derived factor 1 encoding gene, SDF1-3'A, was 0.2125 (95% confidential interval, 0.1608-0.2642). Both values were slightly higher than those obtained earlier for Western European countries. This result can be explained by higher proportion of Asian immigrants, characterized by higher frequencies of these mutations, in the population of Moscow. PMID- 11898621 TI - [Polymorphism of glutathione S-transferases M1 and T1 in several populations of Russia]. AB - Frequencies of the wild-type and null genotypes of the GSTM1 and GSTT1 genes were established in healthy donors from several Russian populations (ethnic Russians from the towns of Oshevensk and Kholmogory, Arkhangel'sk oblast; ethnic Khants; ethnic Kalmyks; and ethnic Buryats) in order to identify the ethnic group with the maximal frequency of the null genotype. The highest frequency of individuals with the null genotype of both genes was observed in the Kalmyk and Buryat populations. The results may be used to study the effect of climatic and ecological conditions on multifactorial disease incidence in populations. PMID- 11898622 TI - [Study of the association between constitutional exogenous obesity and polymorphism of the apolipoprotein B gene]. AB - An attempt was made to associate the insertion-deletion (Ins/Del) polymorphism of the apolipoprotein B gene (apoB) with obesity and to identify alleles and genotypes predisposing to this disorder. The apoB Ins/Del allele frequencies observed in the Russian population were similar to those in West European populations and significantly differed from frequencies reported for Asian populations. Patients with obesity did not differ from healthy individuals in allele and genotype frequencies regardless of whether total or sex-stratified samples were compared. Estimation of relative risk for individuals with genotype Ins/Ins did not reveal a significant association between obesity and this genotype. Thus, constitutional exogenous obesity did not prove to be associated with the Ins/Del polymorphism of the apoB gene in the Russian population. PMID- 11898624 TI - [Russian Scientific Conference "Medical aspects of radiation and chemical safety" (St. Petersburg, Academy of Military Medicine, October 11-12, 2001)]. PMID- 11898623 TI - [Comparative analysis of the effectiveness of countermeasures in rural settlements in the long-term period after the accident at the Chernobyl AES]. AB - A comparative analysis of the strategy of countermeasures aimed at reducing exposure doses to the rural population is presented. The effectiveness of the proposed system of countermeasures assessed by criteria such as reduction factor of exposure doses and cost of averted doses. Costs of countermeasures in the rural settlements located in the affected zone are calculated for a long term after the Chernobyl NPP accident. PMID- 11898625 TI - [Study of stress reactivity in offspring of first-generation rats after irradiation of one or both parents]. AB - Three-month old white mongrel rats (F0) were exposed to X-rays (single exposure dose 1 Gy). Their offsprings F1 were subjected to acute hypokinezia for 60 minutes at different periods of life. The changes of the hormonic status were revealed in males F1; in females F1 deviations were not observed. The stress reactivity of males F1 depended on which of the parents was exposed to radiation. After irradiation of both parents or males F0 only the hyporeactivity in senile age was more pronounced their in control animals of the same age. After irradiation of females F0 only a response to stress in male offsprings F1 corresponded to the control values. PMID- 11898626 TI - [Secretory activity of adrenal glands in the offspring of white rats in conditions of low-intensity radiation effects]. AB - It has been shown that in offsprings of both sex F1, one or both parents (F0) of which were exposed to X-ray radiation at a dose of 1 Gy (the dose rate was 0.4 Gy/min), and in offsprings of 6 generations (F1-F6), parents (F0) of which were placed in to the Chernobyl zone at a pubertal age, the hypertrophy and hyperactivity of isolated adrenals were observed in the different periods of life. The decrease of adrenals' reactivity in response to ACTH (0.1 ED/ml) was found in males of all generations. Most expressed damages of adrenal glands' reactivity were marked in males F6. PMID- 11898628 TI - [Hypothesis of the mechanism of adaptive response induction in mammalian cells by low-dose irradiation]. AB - The mechanism for radiation-induced adaptive response (RAR) in mammalian cells is presented in this paper. The start point of the RAR in the frame of this mechanism is the receptors for growth factors activation due to the increase in the microviscosity of plasma membrane subjected to oxidative damage. There are components of the mitogen-activated signal transduction pathway which take part in the subsequent processes. The main of them are protein kinase C (PK C), motogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK). They make posttranslation modification of the DNA metabolism enzymes and of the transcription factors p53 and c-Jun/AP-1. There are genes taking part in the excision repair and apoptosis among the c-Jun/AP-1 and the p53 targets. C-Jun/AP 1 and p53 can be direct participants at the stage of exision repair when DNA damage is recognized. Thus, the proposed scheme of events removes the contradiction between two hypotheses which explain the RAR: intracell DNA repair induction, either of cell selection in culture of mammalian cells. PMID- 11898627 TI - [Specific long-term cellular changes under effects of low doses of radiation]. AB - We examined the peculiar form of a tissue postirradiative reaction characterizing by massive, dose-independent transition of cell populations to the steady state modification with the essential raise of cell damage and cell loss probability as compared with the probability level of the same alterations in controls. We described some other signs of such type of cellular transformation. It was found that the indicated cellular condition occurred both in active and slowly proliferating tissues. The reaction occurred at relatively low doses of irradiation. Some nonmutagenic factors also may evoke such effects. Our experimental data allow us to suppose the epigenetic mechanizms taking part in the induction and preservation of such alterations. The discovered form of cellular reaction manifestating in different biological objects may be considered as some general biological tendency. The importance of the studied reaction in the pathogenesis of late consequences of low dose irradiation is discussed. PMID- 11898630 TI - [Changes in functional activity of the synthetic apparatus of rat thymocytes under acute and chronic gamma-irradiation]. AB - The changes in functional activity of rat thymocyte synthetic apparatus (synthetic activity) under acute (7.5 Gy) and continuous (dose rates 14.4 and 0.43 cGy/day) gamma-irradiation were studied by the fluorescent microspectral analysis. It has been shown that after the acute irradiation the changes in synthetic activity occurred in three main stages. The stages reflect the depression and activation of synthetic processes that is due to interphase and reproductive cell death and urgent recovery of thymus cellularity and secondary repopulating. Under continuous irradiation with a dose rate 14.4 cGy/day in long term period both the decrease of thymocyte synthetic activity (in most animals) and activation (in the animals with pronounced symptoms of radiation damage) were observed. This reflects the depression processes in immune system and augmentation of immunoreactivity due to mass antigen influence of transformed cells and infectious agents on thymocytes. Under low dose ionizing irradiation (dose rate 0.43 cGy/day) the undulating changes in synthetic processes in thymus cells were observed. This depends on the recurrence of depression and recovery processes in the blood-forming tissue. PMID- 11898629 TI - [Effects of gamma-irradiation and thyro-parathyroidectomy on structural functional state of membranes of sarcoplasmic reticulum of skeletal muscle]. AB - It has been found that the combined effect of the acute gamma-irradiation with 1 Gy dose and thyroparathyroidectomy (TPTE) changes the structure-functional state of SR membranes of rat skeletal muscles conditioned by the disturbance of hormonal regulation of molecular-cellular mechanisms of calcium exchange. PMID- 11898631 TI - [Emotional stress and problems of radiation medicine]. AB - The influence of emotional stress on behaviour and industrial activity of man as well as on the development of posttraumatic stressful frustration and other disorders is considered in connection with real or possible action of ionizing radiation, and on the course of radiation injuries. It is shown that a problem of emotional stress and radiation safety are closely connected. The study is important for solving problems facing emergency medicine. PMID- 11898632 TI - [Recovery of yeast cells after exposure to ionizing radiation and hyperthermia]. AB - Quantitative regularities of recovery of wild-type diploid yeast cells irradiated with gamma-rays (60Co) simultaneously with exposure to high temperatures were studied. It was shown that in conditions of such a combined action the constant of recovery did not depend on the temperature at which the irradiation was carried out. However, with an increase of acting temperature an augmentation in the portion of irreversible component was registered. The analysis of cell inactivation revealed that the augmentation of the irreversible component was accompanied by a continuous increase of cell killing without any postirradiation division after which cells are incapable of recovery. The reproductive death was mainly exerted after ionizing radiation applied alone while in conditions of simultaneous thermoradiation action the interphase killing (cell death without division) predominated. It is concluded on this base that the mechanism of synergistic interaction of ionizing radiation and hyperthermia may be related with cardinal change in mechanisms of cell killing. PMID- 11898633 TI - [Study of mechanisms of anti-irradiation effects of interleukin-1beta in long term bone marrow cultures]. AB - The influence of betaleukin (human recombinant interleukin-1 beta) on the processes of postirradiation recovery of haemopoietic precursors (GM-CFC) and the level of granulocyte-macrophag colony-stimulating factor (GM-CSF) were studied in long-term bone marrow cultures after gamma-irradiation with a dose 2 Gy. Then the betaleukin action on the contents of GM-CFC and induction of GM-CSF in the non irradiated cultures was studied. It was shown that betaleukin increased the induction of GM-CSF and raised the contents of GM-CFC in long-term bone marrow cultures, and the maximal increase of a GM-CSF level and GM-CFC amount was marked in 20 hours after introduction. At an irradiation of long-term bone marrow cultures in conditions of betaleukin introduction 20 hours prior to influence of radiation the smaller degree of damage and faster recovery of GM-CFC was observed. The data in this report suggest that one of the mechanisms of antiirradiation action of betaleukin apparently is connected to the action of the preparation on hematopoietic microenvironment cellular elements, that causes the release of a colony-stimulating factor and stimulation of recovery of haemopoietic precursors. PMID- 11898634 TI - [Dependence of the therapeutic effectiveness of interleukin-1beta on the time of the drug administration after irradiation of mice]. AB - In experiments on mice F1(CBA x C57BL/6) the dependence of 30th days survival on the time of betaleukin (medicine form of interleukin-1 beta) administering after exposure to 7.5 Gy whole body gamma-irradiation from 137Cs (approximately DL80/30) was studied. Betaleukin was injected subcutaneously in dose 25 mcg/kg 0.2, 1, 3, 6 and 24 h after the exposure. The highest therapeutic effect took place in case of 0.2 h interval, then it dropped but was slightly expressed at 1 h. The hemopoiesis condition was studied in 7 or 9 days after the mice exposure to 6 Gy and betaleukin administering 1 h later in dose 25 or 50 mcg/kg. The positive effect on granulopoiesis beginning from the level of CFU-GM was observed. There were analyzed the reasons of weaker betaleukin effect and shorter period of its effectiveness after exposure in comparison with literature data regarding IL-1. PMID- 11898635 TI - [Postradiation recovery of hematopoiesis in mice affected by Neogen (IEW)]. AB - The effect of the synthetic peptide IEW (Neogen) with immunomodulating properties on postradiation recovery of haemopoiesis was investigated. We have shown that Neogen is a potential stimulator of haemopoiesis. The administration of Neogen after irradiation shortened duration of period of the recovery of the compartment of CFU-S-8 and the amount of bone marrow cells. The comparision of the effects of Neogen and GM-CSF (Leucomax) and G-CSF (Granocyte 34) have shown that the targets for these agents are probably different: polypotent CFU-S-for Neogen, and CFU-GM for GM-CFS. Based on the results, we suggested the mechanism of Neogen effects on heamopoiesis. PMID- 11898636 TI - [Experimental study of the role of blocking 5-HT3-receptors of serotonin and D2 receptors of dopamine in the mechanism of early radiation vomiting in dogs]. AB - The experiments on dogs exposed to 137Cs gamma quanta at doses of 8 and 20 Gy showed that Latranum, a selective blocker of serotonin 5-HT3-receptors, is a more efficient antiemetic than Dimetpramidum, a D2-dophamin lytic. This is suggested by fewer animals with emetic reaction or by less severe vomiting in case they have any. The results, taking account of earlier data obtained in the experiments on M. fascicularis monkeys, show that the antiemetic effect of the blocker of serotonin 5-HT3-receptors Latranum is not species-specific and equally well manifests itself in animals with different constitutions of the chemoreceptor trigger zone. PMID- 11898637 TI - [Study of the possibility of hyperparathyroidism development in rats exposed to internal irradiation by iodine radioisotope]. AB - The possibility of hyperparathyroidism development secondary to earlier internal irradiation with radioactive iodine was studied experimentally in Wistar rats. This report describes the parathyroid morphology and biochemical findings for animals irradiated with 131I at the doses of 4.5, 40, or 80 Gy. The interval between the radiation exposure of two-month-old rats and their examination for thyroid and parathyroid pathology was 14 months. Neither hypercalcemia nor hypophosphatemia was found. Moreover, the level of calcium in serum slightly decreased following 40 and 80 Gy irradiation. The increased incidence of parathyroid fibrosis and hypofunctional structure transformation were revealed. PMID- 11898638 TI - [Effects of radiation-ecological conditions and experimental hyperthyroid states on indicators of testis and prostate androgen receptors in rats]. AB - Dose-dependent changes of the molecular characteristics on androgen receptor (AR) systems in gonads of male rats were studied at experimental L-thyroxine-induced states after low doses irradiation exposures in different reference points of 10 km Chernobyl zone. The data obtained suggest a generalized working mechanism of "oscillatorous changes" for contents, affinities and cooperative properties of AR to its natural ligands as a "mirror" reflecting some adaptational reactions in target cells that modulates their androgen-controlled biochemical activity. PMID- 11898639 TI - [Teratogenic effects of incorporated radionuclides]. AB - Experimental data on teratogenic effects induced by incorporated alpha, beta and gamma-emitters were analyzed. It was found that the radioactive substances as well as external irradiation induced teratogenic effects. Teratogenesis caused by incorporated radionuclides has some peculiarities compared to the effect caused by fetus exposure to external radiation. These peculiarities are related to the fact of the limited penetration of incorporated radionuclides via placenta barrier so the radiation fetal doses are accumulated within long period of time and radiation dose rates are relatively low. The exposure to incorporated radionuclides does not induce severe developmental defects. Most frequent developmental defects of fetus include its death, general retardation of the development and growth. In such case the earlier pregnancy term was affected by radionuclide the more severe fetal damages occur in fetus because of the gradual increase of absorbed dose even in case of single intake of radionuclide. RBEs of radionuclides if compared to that for external gamma radiation are evaluated as follows: 2-4 (tritium oxide), 20 (241Am), 50 (238Pu) and 3-5 (131I in thyroid). PMID- 11898640 TI - Demand for dollars going up. Updated demographics support need for increased capital. PMID- 11898641 TI - Restorative care: when doing less can help more. PMID- 11898642 TI - A new economic era. Seniors' changing financial status will set the pace. PMID- 11898643 TI - Your place or theirs? Facility based providers are bringing the nursing home. PMID- 11898644 TI - What lurks under the sink? Chemical safety is a vital part of staff education. PMID- 11898645 TI - Helping families get loved ones to day care. Caregivers need to be prepared to deal with the transition. PMID- 11898646 TI - Chasing efficiency. With clinical, financial and research priorities in mind, hospital CFOs expect and demand the most from financial applications. PMID- 11898647 TI - No Catch-22 here. For 22 Connecticut hospitals, using a uniform Web-based decision support system has cut operating costs and streamlined business. PMID- 11898648 TI - Automating procedure workflow to boost productivity. Endoscopy center automates an inefficient manual workflow and increases patient volume. PMID- 11898649 TI - Managed care grows up. NMHCC convention to focus on new solutions. PMID- 11898650 TI - Toward a unique patient identifier. Florida IDN attacks duplicate records with MPI software, consultation and a shift in organizational philosophy. PMID- 11898651 TI - What works: wireless. Access on the go. PMID- 11898652 TI - Financial systems HotList. PMID- 11898653 TI - Column fodder. PMID- 11898655 TI - [Method and construction of programmed electric muscle stimulation in gait disorders]. AB - The Research Institute of Prosthesis and Prosthesis Making, Moscow, has developed a method for artificial movement correction (AMC) and designed a programmable muscle electrostimulation device (PMED) for its application. The essence of the AMC method is that to use a multichannel PMED in a good agreement with the natural programme of muscle excitation and contraction in the locomotor cycle. Due to this, many days' training enhances the viability of muscles, corrects movements, and gives rise to near-normal gait. The AMC method was used to design stationary and portable electro-stimulators synchronized with step phases wherein muscles function. Among other things, an 8-channel movement correcting apparatus controlled from a personal computer, which is adjustable to the walking rate has been designed. The AMC method has shown a high therapeutical effect in over 4500 neurological and orthopedic patients. PMID- 11898654 TI - [Prospects for using consecutive rank processing for developing portable cardiac devices]. AB - The paper deals with detection of the QRS complex of a cardiac signal at the level of interferences by the rank method. An algorithm is proposed to detect informational cardiac pulses of an electrocardiographic signal and its temporary ranges. The use of such algorithms provides portable devices to examine an electrocardiographic signal when the patient freely moves under high interference. PMID- 11898657 TI - [Method for gas-discharge visualization and automation of the system of realizing it in clinical practice]. AB - The stage of diagnosis is essential in terms of both the choice of the most adequate treatment policy and the monitoring of human responses to various environmental factors, including living and working conditions, conflicts, predisposition to some diseases, etc. To develop new non-invasive automatic procedures that assess the body's condition during minimum interventions into vital functions is of great scientific and practical value. The gas-discharge visualization method is a promising direction in the construction of these procedures. PMID- 11898656 TI - [Mathematical modeling of the process of thermal action of the surface of human skin]. AB - The mathematical model of the physical system, which represents a pattern of a pulse heat source and a skin surface portion, is regarded as a dynamic process of thermal action on man. The model reflects a relationship of the parameters of a thermal pulse, the threshold characteristics of a thermoreceptor, and the thermal parameters of the skin. PMID- 11898658 TI - [Information-computer modeling of suppression of the infectious tuberculosis process through sterilization of medical instrumentation and equipment by using electrochemically activated aqueous solutions]. AB - The paper deals with the topical problem in the informational computer simulation of suppression of an infectious tuberculosis process by applying electrochemically activated aqueous solutions in order to sterilize medical instruments and equipment. The efficiency of use of electrochemically activated aqueous solutions for sterilization of medical instruments and equipment and for disinfection of other contaminated objects was evaluated by an computer-aided experiment and a retrospective analysis of the epidemic situation by using the original mathematical models of infectious processes by taking into account the aspects of medical engineering technology. PMID- 11898659 TI - [Analysis of the market for medical instruments based on patent statistics]. AB - The paper shows the role and value of patent statistics under the present conditions, analyzes the general economic situation in Russia and the structure of the medical instrument making market by using patent statistics. PMID- 11898660 TI - [Formal model of automating a system for prophylactic screening of the population]. AB - Experience in designing and using screening diagnostic systems has yielded a formal model of an automatic system to make a prophylactic surveys of the population. This model makes it possible to design automatic systems so that a number of informational structures should be developed to store initial medical data and the results of their processing, to aggregate decisive rules of making a conclusion on the health status and the relatively invariant interpreting core. The similar approach substantially simplifies a procedure to modify decisive rules from the results of using the system and to duplicate off-the shelf solutions on different quota of patients with minimal changes in software implementation. PMID- 11898661 TI - [Polymer products for anesthesiology and intensive therapy]. AB - The paper enlists medical products made from polymer materials, which are manufactured by the Metom RNCF. It describes their properties and gives the nomenclature of transparent facial masks used under inhalation anesthesia and artificial lung ventilation. The types and basic data on respiratory polymer hoses are reported. PMID- 11898662 TI - [Electronic handgrip dynamometer]. AB - The paper outlines an electron handgrip dynamometer designed by the authors by taking into account the requirements for both latest and obsolete electron dynamometers. It also considers the comparative characteristics of currently available electron handgrips. PMID- 11898663 TI - [Computer biocontrol systems: trends in development]. AB - The paper considers the basic trends in the development of the new computer technology, such as biocontrol based on the analysis of its development in the past 10 years. New variants of biocontrol, new advances in engineering and use are noted. Emphasis is laid on the new branch of biocontrol computer remedial games. PMID- 11898664 TI - [Experience in using systems for automating endoscopic diagnosis]. AB - The specific features of processes of current methods for instrumental diagnosis, which are used at therapeutical-and-prophylactic institutions are considered. Main problems in automation of endoscopic diagnosis and the current methods of their solution are discussed. Tasks required to put an automatic diagnostic system into practice are defined. The results of use of one of the developed variants of the system that has particular features of its implementation are presented. Sources of the efficiency of such systems are shown. PMID- 11898665 TI - [Development and experience in clinical use of computer monitoring systems in the cardiology intensive care unit]. AB - To develop national monitor computer systems (MCS) for intensive cardiological care units, which should be reliable, easy-to-use, and cheaper than their foreign analogues is a pressing scientific and practical problem for Russian medical instrument making. The paper presents the results of development of and clinical experience with MCS at the intensive cardiological care unit of the Republican Clinical Cardiology Dispensary in the Republic of Udmurtia (Izhevsk). PMID- 11898666 TI - [Experience in using the "Lancet" laser device in surgical practice]. AB - The paper deals with the authors' experience in using new-generation Lantset laser surgical apparatuses that show a high spectral radiation purity, a high radiation coherence, continuous and pulsed operations. It also presents their characteristics and description, the results of their clinical application. PMID- 11898667 TI - [Status and prospects for development of medical equipment at the state-of-the art "Axion Holding" "Izhevskiy Motor Plant"]. AB - The joint stock company "Izhevsky Motor Plant "AXION-HOLDING" is a large manufacturer in different industrial branches, including medical equipment. The paper presents data on the status-of-the-art and prospects of development in producing medical equipment in several areas, including information on specific articles that are manufactured and promising. PMID- 11898669 TI - Management of acne. PMID- 11898668 TI - [Development of computer technology for increasing the resolution of radiographic images in mammography]. AB - A computer technology has been developed to enhance the resolution of an X-ray signal by 1.5-2 times than the baseline values. The results have been tested on real mammological X-ray films that are to detect minor inclusions, such as calcinates. PMID- 11898670 TI - Practice patterns related to mechanical restraint use across a multi institutional health care system. AB - In recent years, regulatory and governmental initiatives have focused increased scrutiny on the use and practices associated with mechanical restraints. Consequently, hospitals are increasingly measuring and comparing both internally and externally their restraint practices as they strive to optimize their use and assure the safe care of patients being restrained. This study analyzes 12,860 restraint episodes from 10 acute care hospitals in a single health care system. Overall findings support many previously identified trends related to the types of restraints used and reasons for application. However, findings from this study also suggest that there are differences among rural, community, and tertiary hospitals. This study also provides the first widespread documentation of rates and types of alternative methods attempted and common patient care practices carried out during restraint application. These results can serve as external comparisons for other acute care settings as they strive to minimize and assure safety in restraint application. PMID- 11898671 TI - Integration of pressure ulcer treatment protocol into practice: clinical outcomes and care environment attributes. AB - The purpose of this study was to determine if a research-based protocol for pressure ulcer treatment that had been successfully implemented in a long-term care facility was sustained over time. A secondary aim was to describe the attributes of the care environment that may have contributed to or impeded the maintenance of this protocol. A retrospective chart review was conducted of all patients who developed incident stage II, III, or IV pressure ulcers in the facility over a one-year period five years after initial implementation of a pressure ulcer treatment protocol. Data regarding ulcer characteristics and type, frequency, and duration of treatments were collected. Subjects were followed until the ulcer healed, the subject died or was discharged, or the 1-year study period ended. Care environment attributes, including patient care hours, turnover and stability rates, salaries, decision-making structures, and facility mission were obtained from the facility's Human Resource Department and existing databases in the Nursing Services Department. Outcomes of protocol implementation were defined as ulcer healing and costs associated with treatment. Costs were calculated from the provider perspective and included cost of supplies and labor consumed in providing direct pressure ulcer care. Of the 46 incident ulcers treated during the one-year study period, 40 (87%) healed and five (11%) were unhealed when the subject died. One ulcer remained unhealed at the end of the study. The total cost for treatment of these incident ulcers was $18,688, with nursing labor comprising 80% of the total expenditures. Adherence to the protocol, which contained predominantly inexpensive moist wound healing treatment options, resulted in complete healing of most pressure ulcers at a relatively low cost to the facility. The organizational environment of the facility, which maintains staffing levels and salaries at higher than national averages and promotes staff nurse accountability and decision making, may have provided the necessary climate to overcome barriers to clinical integration and sustain the desired care practices. PMID- 11898673 TI - Recovery from coronary artery bypass surgery: age-related outcomes. AB - As people age, their incidence of coronary heart disease increases. The majority of persons undergoing invasive procedures such as coronary artery bypass surgery are 65 and older. Because of population trends related to aging, it is projected that there will be exponential increases in the numbers of people requiring treatment for this health problem in the future. Changes in health care reimbursement have significantly decreased hospital length of stay, resulting in many patients completing their recovery either in a rehabilitation facility or at home. Patients with multiple preoperative comorbidities are at risk for postoperative complications. Older patients usually have more health problems after coronary artery bypass surgery because they have more risk factors prior to the procedure. It is not known whether there are differences in outcomes between comparatively older and younger patients when they are matched by risk classification. Information on the recovery of patients at home will enable nurses to meet their care needs prior to surgery and after discharge from the hospital. PMID- 11898672 TI - Factors predicting 12-month outcome of elderly patients admitted with hip fracture to an acute care hospital. AB - Elderly patients (n = 121) with hip fracture were followed to determine if: (1) outcomes measured 12 months post-fracture differed significantly from pre fracture measures, and (2) patient characteristics on hospital admission predicted three outcomes (site of residence, function, and walking status) 12 months later. At 12 months fewer patients resided at home. They had declined functionally. Baseline cognition, residence site, function, and walking individually predicted outcomes. However, outcomes were predicted best by multiple variables. These findings can be used to educate patients, their families, and the public on outcomes and their determinants after hip fracture. PMID- 11898674 TI - Using computer simulations and focus groups for planned change in prenatal clinics. AB - The Colleges of Nursing and Engineering in a southwest Florida university combined efforts to design a project to use time/motion techniques and focus groups to assess patent flow and effective and efficient use of human resources in public health clinics. Data for 877 observations were entered into a computer simulation program that displayed alternative configurations for health resource management. Information from focus groups was used to plan for ways to use clinic wait time more effectively. This article describes data collection and findings. PMID- 11898675 TI - Health-seeking behavior as an outcome of a homeless population. AB - Health care is an ever-present concern for homeless individuals. Health-Seeking Behaviors within this population are examined from a nursing perspective. Complex Relationship Building, considered essential in addressing Health-seeking Behaviors for the homeless, is examined in regard to access, trust, and follow up. It is believed that access and follow-up are related to convenience of site and the matter of trust. A pilot study of how to measure the concept of trust was conducted. Factor analysis for the Gibson Trust Instrument shows two factors: (1) interpersonal attributes and (2) behavior attributes. The Gibson Trust Instrument can be used to measure trust, which is crucial for Complex Relationship Building. PMID- 11898677 TI - Clinical pathways and guidelines for care management. PMID- 11898678 TI - Measuring health care outcomes with secondary data. PMID- 11898679 TI - Defibrillator/monitor/pacemakers. AB - Defibrillator/monitors allow operators to assess and monitor a patient's ECG and, when necessary, deliver a defibrillating shock to the heart. When integral noninvasive pacing capability is added, the resulting device is referred to as a defibrillator/monitor/pacemaker. In this Update Evaluation, we present our findings for one newly evaluated model, the Philips Heartstream XL, and we summarize our findings for the seven previously evaluated models that are still on the market. (Our previous Evaluations were published in the May-June 1993, February 1998, and September 2000 issues of Health Devices.) Defibrillator/monitor/pacemakers are used for a variety of applications within the hospital, as well as by emergency medical services (EMS) personnel and others in the prehospital environment. To help both hospital-based and prehospital users select an appropriate model, we rate the models (1) for each of three in-hospital applications--general crash-cart use, in-hospital transport use, and in-hospital use by basic as well as advanced users--and (2) for prehospital (EMS) use. For in hospital use, we recommend four of the evaluated models. These received either Preferred or Acceptable ratings for all the applications considered. For prehospital use, we found that five of the models will meet most organizations' needs. PMID- 11898680 TI - Speech recognition systems. Are they up to the task? AB - Clinical speech recognition systems record speech input from a physician and translate the acoustic data into text output that forms the basis of medical reports. These reports can then be edited by physicians either during or after dictation. Speech recognition systems have received a lot of attention as potential time-savers. But when we looked at four of these systems, we found their accuracy to be low enough that it will take most physicians 20% to 30% more time to produce a report than is required using traditional transcription methods. Even so, these systems offer enough benefits that you should at least investigate their use. For one thing, they're considerably less expensive than using a transcription service. For another, despite the additional physician time required, a report can be turned around far more quickly than with transcription- a matter of hours rather than days. With proper planning, most facilities should be able to implement these systems successfully. PMID- 11898681 TI - Not for children: most automated external defibrillators will deliver too much energy to pediatric patients. PMID- 11898682 TI - Electrosurgery on patients with implantable cardioverter-defibrillators. PMID- 11898683 TI - Speech recognition systems can substitute the wrong drug name. PMID- 11898684 TI - Companionship, loss, and taking risks. A mosaic of applied holism. AB - This article describes personal and professional encounters in the life of a nurse who reflects on holism and its place in these experiences. In an attempt to grow and care for ourselves and others, holistic principles such as intention, caring presence, attention to environment, and rituals can provide structure, inspiration, and comfort. PMID- 11898685 TI - The world has never needed holistic nurses more than now. PMID- 11898686 TI - Weaving the fabric of spirituality as experienced by patients who have undergone a coronary bypass surgery. AB - When one is faced with a life-threatening event, spiritual issues become extremely important. The depth of one person's suffering is a unique experience. The relationship that developed between the interpretation of the answers of the coronary artery bypass patients and self-interpretation were the two directions in which this hermeneutic research moved forward within the hermeneutic circle. Central implications of the concept of spirituality emerging from this study are spirituality as meaning through gaining a new appreciation of life and health, spirituality as an inner strength through love and faith and in an ontological sense, spirituality as an inner strength through suffering and desire. Suffering invites one into the spiritual domain. Indeed, this concept raises important considerations for the development of caring knowledge about spirituality. PMID- 11898687 TI - The effect of an educational intervention on willingness to receive therapeutic touch. AB - The purpose of this quasi-experimental study was to test the effect of an educational intervention on participants' willingness to receive Therapeutic Touch (TT). A sample of 108 participants was recruited from three area nursing programs and one professional business women's group. Participants completed questionnaires before and after a 10-minute talk on TT and a 5-minute demonstration of TT. Participants were asked to rank their willingness to experience a TT treatment and to explain the reasons for their rankings. The hypothesis that the intervention would increase participants' willingness to experience TT was supported; there was a significant difference in pre- and postintervention willingness to experience TT. PMID- 11898688 TI - Spirituality in persons with heart failure. AB - Spiritual expression has been proposed as a dimension of quality of life. Persons with chronic diseases such as AIDS or cancer have described the value of spiritual expression in living with their illnesses. The authors examined the role spirituality plays in the lives of 58 people with heart failure being treated medically or by transplant. Instruments used included the Medical Outcome Survey Short Form 36 and Index of Well-Being measures of quality of life, the Spiritual Well-Being Scale, and the Relative Importance Scale. Combined spirituality scores predicted 24% of the variance in global quality of life. There were no significant gender differences in spiritual well-being or quality of life. PMID- 11898689 TI - The effect of an integrated stress management program on the psychologic and physiologic stress reactions of peptic ulcer in Korea. AB - The purpose of this study was to identify the effects of an integrated stress management program on symptoms of stress and ulcer healing in a sample of Koreans at a major medical center in Seoul. The study employed an experimental design with two treatment groups. One treatment group (n = 23) participated in an integrated stress management program (ISMP) that consisted of seven 1-hour sessions over a 4-week period. A second treatment group (n = 24) was only given a tape on progressive muscle relaxation (PMR). The ISMP treatment group reported significantly lower stress symptom scores than the PMR only group (t = 3.66, p < .001). The ISMP group also demonstrated greater improvement in ulcer healing than the PMR group (t = 1.95, p < .05). The integrated stress management program was more effective in decreasing self-reported stress symptoms and resulted in more significant ulcer healing than the progressive muscle relaxation treatment. PMID- 11898690 TI - Uncovering hope with clients who have psychotic illness. AB - This article presents findings about how community mental health nurses enable young adult clients who have schizophrenia uncover hope for the future. Grounded theory methodology and simultaneous data collection and analysis took place using interviews and observations. The study was undertaken in the community in regional and rural New South Wales, Australia, and the participants were clients, significant others, and community mental health nurses. The study findings show that nurses used two main strategies to uncover hope: enhancing motivation and developing pathways to wellness. These activities have to be seen within the context of good nurse-client relationships, advancing client self-determination, education, and planning for the future. The process of uncovering hope can be compromised for various reasons. The findings highlight the importance of hope for clients with schizophrenia and the way in which nurses can enable the process. PMID- 11898691 TI - Possible improvement of the clitoral and vaginal blood flow using a somatostatin analog in chronic spinalized Sprague-Dawley rats. AB - We evaluated the effect of a somatostatin analog (octreotide) on clitoral and vaginal blood flow following suprasacral spinal cord injury (SCI) in rats. Twenty four spinalized female Sprague-Dawley rats were randomized into 4 equal groups. The first group served as control paraplegics. The other three groups received octreotide (60 micrograms/day/4 weeks) immediately, 2 weeks, and 4 weeks following SCI. At the end of the experiment, a laser Dopper was used to measure blood flow changes following clitoral and pelvic nerve plexus stimulations. Marked decreases in both clitoral and vaginal blood flow in the control paraplegics were recorded. Significant increases (p < 0.05) in both clitoral and vaginal blood flow were recorded in animals that received octreotide; however, the increase was marked in the animals that received the drug immediately following SCI. Improvement in the clitoral and vaginal blood flow of spinalized rats using octreotide indicates that octreotide may be helpful for patients with SCI. PMID- 11898693 TI - Regulatory and drug development issues related to female sexual dysfunction. AB - Following the approval of sildenalfil for the treatment of erectile dysfunction, an increased awareness of and interest in female sexual dysfunction developed on the part of the academic and research communities as well as the pharmaceutical industry. This article will focus on regulatory issues related to the development of drug products to treat female sexual dysfunction and will describe a recently published drug development guidance document for this indication. PMID- 11898692 TI - Neurophysiology and pharmacology of female genital sexual response. AB - Vaginal sexual arousal is a vasocongestive and neuromuscular event controlled by facilitatory parasympathetic and inhibitory sympathetic inputs. Autonomic preganglionic parasympathetic and inhibitory sympathetic fibers to the vagina and clitoris originate in the spinal cord in the sacral parasympathetic nucleus at the sacral level and in the dorsal gray commissure and the intermediolateral cell column at the thoracolumbar level, respectively. Parasympathetic fibers are conveyed by the pelvic nerve, and sympathetic fibers are conveyed by the hypogastric nerve and the paravertebral sympathetic chain. The activity of these spinal nuclei is controlled by descending projections from the brain and sensory afferens (conveyed in the pudendal, hypogastric, pelvic, and vagus nerves) from the genitalia. A key but unresolved issue concerns the neurotransmitters involved in the control of genital sexual arousal. At the peripheral level, acetylcholine plays a minor role in the regulation of vaginal blood flow, however, recent data suggests that it may be involved in the control of vaginal smooth muscle contractions. Vasoactive intestinal peptide and nitric oxide may be responsible for the increase in vaginal blood flow during sexual arousal, whereas noradrenaline is likely inhibitory. Within the central nervous system, serotoninergic projections from the brain to the spinal cord likely inhibit the induction of genital arousal by peripheral informations (spinal reflex). Although some neurotransmitters regulating the display of sexual behavior have been identified (for example, dopamine), their involvement in the control of genital sexual arousal has not been invested. Anatomical and electrophysiological data point to a contribution of the paraventricular nucleus of he hypothalamus and the median preoptic area, respectively, as key elements in the control of genital arousal. The recent development of models allowing the assessment of vaginal sexual arousal in anesthetized female rats should assist in deciphering the neurochemical pathways controlling vaginal sexual arousal and the development of suitable pharmacological treatment for female sexual dysfunctions. PMID- 11898694 TI - Clitoral pain: the great unexplored pain in women. AB - Clitoral pain is not often reported by patients or in literature. The author reports on 7 women from his practice and 14 women who were on-line volunteers, all of whom had clitoral pain as a major feature. Features included mild to moderate rest pain and significant contact, light-touch induced or pressure induced pain. Associations include Multiple Sclerosis, GUillain Barre Syndrome, urethral sphincter dyssynergia, various vulvar pain syndromes (nine cases), post hysterectomy, Lichen Sclerosis (five cases), spondylolisthesis, vaginismus and genital or pelvic trauma. Eight claimed painful, allodynic nipples and seven had sensitive nipples. Clitoral pain was an important cause of pain in these women. PMID- 11898695 TI - Decreased free testosterone and dehydroepiandrosterone-sulfate (DHEA-S) levels in women with decreased libido. AB - A prior study has shown that premenopausal women could have decreased testosterone levels and still have regular menstrual cycles (Guay, 2001). Since ovarian function in such women was normal, the question of a possible adrenal dysfunction causing androgen deficiency was considered. If this was true, the question then arose as to whether the same defect could be seen in postmenopausal women. We studied 105 women who presented during a 6-month period of time with the chief complaint of decreased sexual desire. On subsequent testing, 74 of the women (70%) were found to have decreased total testosterone, free testosterone, and dehydroepiandrosterone sulfate (DHEA-S). Thirty-six of these women were premenopausal (ages range 24-50 years), and 38 were postmenopausal (ages range 47 78 years). All androgen levels for the women were lower than age-matched control groups found in the literature. The decreased DHEA-S levels suggest a a defect in adrenal steroidogenesis, which was seen in both premenopausal and postmenopausal women. Possible defects in regulatory mechanisms of adrenal steroidogenesis are discussed. PMID- 11898696 TI - SSRI sexual dysfunction: a female perspective. AB - Women experience two to three times the rate of depression that men do. Selective serotonin reuptake inhibitors (SSRIs) are prescribed for many conditions other than depression, such as anxiety disorders, premenstrual dysphoric disorder, pain syndromes, impulse control disorders, and personality disorders, some of which are more common in women. Increasing awareness of sexual side effects has tempered the initial enthusiasm with which SSRIs were greeted. Men taking SSRIs report higher rates of sexual side effects than women taking them, however, women seem to experience more severe sexual dysfunction. In this article, we discuss the epidemiology of sexual dysfunction and describe treatments with sildenafil. PMID- 11898697 TI - Validated instruments for assessing female sexual function. AB - In this article, we review five instruments for assessing female sexual dysfunction (FSD): the Brief Index of Sexual Functioning for Women (BISF-W; Taylor, Rosen, & Leiblum, 1994), the Changes in Sexual Functioning Questionnaire (CSFQ; Clayton, McGarvy, & Clavet, 1997), the Derogatis Interview for Sexual Functioning (DISF/DISF-SR; Derogatis, 1997), the Female Sexual Function Index (FSFI; Rosen et al., 2000), and the Golombok Rust Inventory of Sexual Satisfaction (GRISS; Rust & Golombok, 1986). The purpose of this article is to highlight the psychometric properties of these questionnaires in an effort to assist researchers in selecting effective measurement tools for FSD. PMID- 11898698 TI - Androgen replacement therapy with dehydroepiandrosterone for androgen insufficiency and female sexual dysfunction: androgen and questionnaire results. AB - During our evaluations of women with sexual dysfunction, we have seen many with low interest, arousal, and orgasmic capabilities with associated personal distress and diminished genital sensation and blood flow following sexual stimulation. Laboratory evaluation of these women has revealed normal estrogen but androgen values that were either below or in the lower quartile of the physiologic range. Androgen insufficiency and sexual dysfunction have been the working diagnoses in these women. Although many treatment options currently are available for this syndrome, there are limited data concerning safety and efficacy. The aim of this retrospective, Institutional Review Board (IRB)- approved, single-institution study was to report on the androgen and questionnaire results from a series of patients who underwent androgen replacement therapy with dehydroepiandrosterone for treatment of androgen insufficiency and sexual dysfunction. This study revealed that there was a significant decrease in sexual distress, a significant increase in sexual function in the domains of desire, arousal, lubrication, satisfaction, and orgasm, and a normalization to values within the physiologic range in the following androgens measured: total testosterone, free or bioavAilable testosterone, DHEA, DHEA-S, and androstenedione. Side effects included increased facial hair (11%), weight gain (7%), acne (5%), temporary breast tenderness (1%), loss of head hair (1%) and skin rash (1%). Preliminary results suggest that androgen replacement therapy with dehydroepiandrosterone is a safe and effective treatment for androgen insufficiency and female sexual dysfunction. However, further research is needed, including prospective, multi-institution, placebo controlled double-blind studies. PMID- 11898699 TI - Women's sexual desire--disordered or misunderstood? AB - A new model of women's sexual response moves the focus from spontaneous drive with its markers of sexual thoughts, fantasies, and conscious urge to be sexual to an inherently responsive cycle. The model reflects intimacy-based sexual motivation, processing of sexual stimuli to arousal, cognitive, and affective appraisal of that arousal. Sexual desire to continue the physical experience is accessed later. Providing that the outcome is emotionally and physically satisfying, emotional intimacy with the partner is increased. Any spontaneous sexual drive augments this intimacy-based cycle. Analysis of one or many breaks in the cycle has therapeutic implications. PMID- 11898700 TI - Characteristics of female patients with sexual dysfunction who also had a history of blunt perineal trauma. AB - Perineal trauma can occur in both genders, however, data supporting the relationship between sexual dysfunction and blunt perineal trauma in women is lacking. This study reviewed the patient characteristics of women with sexual dysfunction who also had a history of blunt perineal trauma. A neurogenic form of sexual dysfunction has been implicated, with primary complaints of orgasm disorder and abnormalities noted on genital sensory testing. Further research in this area is needed. PMID- 11898701 TI - The prevalence of phimosis of the clitoris in women presenting to the sexual dysfunction clinic: lack of correlation to disorders of desire, arousal and orgasm. AB - Physical examination of the genitalia was performed during an evaluation of women with sexual health problems. Cephalad displacement of the right and left labia minora enables full retraction of the clitoral prepuce and complete exposure of the glans clitoris, under normal circumstances. We defined clitoral examination as abnormal when the cephalad force resulted in varying degrees of incomplete foreskin retraction and limited exposure of the glans clitoris. The pathophysiology is likely to be secondary to recurrent vulvar dermal infections of blunt trauma changing prepucial elasticity. Clitoral phimosis, a previously undiagnosed physical finding, was identified in 22% of the women. Other than its link to sexual pain, the clinical significance of this finding, in particular the relation to diminished sensitivity and impaired orgasmic capability, is unclear at this time. PMID- 11898702 TI - Clitoral artery blood flow in healthy young women: preliminary report on menstrual cycle and hormonal contraception. AB - The aim of the present study was to measure clitoral artery blood flow throughout the menstrual cycle and in oral contraceptive users. We recruited healthy young women (n = 19, age range: 21-28 years; body-mass index: 18-23 kg/m2) without sexual dysfunction (Female Sexual Functioning Index criteria; Rosen et al., 2000). Clitoral arterial peak systolic velocity (PSV) in at least two phases of the same ovulatory cycle or during the second week of the pill was measured by doppler ultrasonography. Clitoral arterial PSV measures (cm/s) were superimposable during the follicular and the luteal phase of the menstrual cycle (10.4 +/- 1.2 versus 10.2 +/- 1.6), whereas a slight but significant increase (12.2 +/- 1.2, f = 3.99; p < 0.04) was evident at the time of ovulation. In addition, PSV measures were significantly higher in women taking hormonal contraception compared to women studied throughout the menstrual cycle (14.2 +/- 2.7 versus 10.8 +/- 1.5; p < .001). Whether or not these preliminary data may be of any significance to female sexual arousal throughout the menstrual cycle remains to be established. PMID- 11898703 TI - FSD partner issues: expanding sex therapy with sildenafil. AB - Sildenafil reinvigorated sex therapy, expanding the number and range of individuals restored to sexual health. Sildenafil used adjunctively with sex therapy accelerated therapy and improved outcome in treating erectile dysfunction (ED). For women, sildenafil initially was used "off label" as a primary treatment for female sexual dysfunction (FSD). However, sildenafil could also be used in conjunction with sex therapy for dysfunctional male partners of women with FSD, so that his ED does not sabotage her treatment. This article describes an integrated treatment, where adjunctive sildenafil use was an important strategic component in the sex therapy of a couple's unconsummated marriage in which the wife's vaginismus, dyspareunia, and anorgasmia was complicated by her husband's ED and retarded ejaculation (RE). PMID- 11898705 TI - Epidemiological characteristics of 250 women with sexual dysfunction who presented for initial evaluation. AB - There have been limited literature reports concerning the epidemiological characteristics of female patients who present for initial evaluation of sexual health problems to an outpatient sexual health clinic. This study is a single institution, retrospective, IRB-approved, observational assessment of 250 female patients undergoing management for sexual dysfunction. In our clinic, women with sexual dysfunction were, in general, young, healthy, and free of vascular risk factors; complained of an acquired multidimensional combination of decreased desire, arousal, and orgasm; and had significantly low serum androgen levels. More research in the management of women with sexual health problems is needed. PMID- 11898706 TI - Beyond dysfunction: a new view of women's sexual problems. AB - A new theoretical framework and classification system for women's sexual problems, written by feminist clinicians and social scientists, was released in October 2000. Part one critiques the current American Psychiatric Association nomenclature for women's sexual programs. Part two highlights international sexual rights documents. Part three begins with a women-centered definition of sexual problems: "discontent of dissatisfaction with any emotional, physical, or relational aspect of sexual experience" (pp. 228-229). It also provides four categories of causes: (a) sociocultural, political, or economic factors; (b) partner and relationship factors; (c) psychological factors; and (d) medical factors. The document is designed for professionals and the public. PMID- 11898704 TI - Phentolamine mesylate in postmenopausal women with female sexual arousal disorder: a psychophysiological study. AB - The objective of this study was to assess the potential of phentolamine as a treatment of postmenopausal women with female arousal disorder (FSAD). Vaginal photoplethismography and a subjective questionnaire were used. Forty one women were enrolled and four treatments were tested: vaginal solutions 5 mg and 40 mg and an oral tablet each of 40 mg of phentolamine and placebo. Physiological readings were significantly different from placebo in the women using hormone replacement therapy (HRT) with 40 mg of phentolamine in vaginal solution (p = 0.0186). Subjective reports also were significantly different from placebo with the vaginal solution 40 mg and the oral tablet of 40 mg of phentolamine among hormone replacement users. No significant differences were found among women not receiving HRT. Results indicate that phentolamine may show promise as treatment for FSAD in estrogenized postmenopausal women. PMID- 11898707 TI - Androgens in female genital sexual arousal function: a biochemical perspective. AB - Female sexual dysfunction is a highly prevalent, multicausal, and multidimensional medical problem that has a major impact on quality of life and interpersonal relationships. It is hypothesized that genital arousal disorder is caused by diminished arterial blood flow within the hypogastric arterial bed. This hypothesis may explain the pathophysiologic mechanisms in older patients with vascular risk factors, but it does not account for either the prevalence or the mechanisms of arousal disorders seen in young women with no vascular risk factors. Hormonal imbalances may be important contributing factors to the pathophysiologic mechanisms involved in female arousal disorders. Androgens modulate the growth and function of female genital sexual organs included the labia, vagina, and clitoris. This article discusses the potential role for androgens in facilitating female genital sexual arousal. PMID- 11898708 TI - Expressions of female sexuality. AB - Past commentaries in scientific literature on female sexuality extrapolate male psychosocial beliefs and anatomical data to the female form. A literature review of journals and books captures descriptive aspects of female sexuality and identifies a unique physiological and psychosocial system. Similarities between the sexes of arousal, desires, and response continue to lack scientifically definable constructs of validity and reliability. Research that validates the differences in women and unifies the documented fragments is imperative to understanding a distinct model of sexual dysfunction and to equalizing the balance of therapeutics offered, which can enhance female sexual expression. PMID- 11898709 TI - Genital and subjective sexual arousal in postmenopausal women: influence of laboratory-induced hyperventilation. AB - The current study was aimed at comparing genital and subjective sexual arousal in pre- and postmenopausal women and exploring the effects of heightened sympathetic nervous system (SNS) activity on these parameters. Seventy-one women (25 young and premenopausal, 25 postmenopausal, and 21 age-matched premenopausal women) participated in two counterbalanced sessions consisting of genital arousal assessment with vaginal photoplethysmography and subjective arousal assessment with self-report questionnaires. SNS activity was enhanced using laboratory induced hyperventilation. Results demonstrated no significant differences between pre- and postmenopausal women on genital and subjective measures of arousal in response to neutral and erotic films. SNS manipulation increased genital excitement only in young, premenopausal women. These data suggest that prior SNS enhancement can differentiate pre- from postmenopausal genital arousal. Data also revealed significant correlations between genital and subjective sexual arousal in older pre- and postmenopausal women, but not in young premenopausal women. These data are the first to directly compare genital-subjective correlations between pre- and postmenopausal women. PMID- 11898710 TI - Sexual dysfunction in type II diabetic females: a comparative study. AB - Diabetes Mellitus (DM) is considered to play a principle role in the etiopathogenesis of sexual dysfunction both in men and women. The aim of this study is to evaluate sexual function in Type II diabetic women. A total of 72 young diabetic women (mean age: 38.8 years) with no other systemic diseases and 60 age-matched healthy women were enrolled in our study. We sought from them a detailed medical and sexual history and used the Index of Female Sexual function (IFSF) questionnaire (Kaplan et al., 1999). The mean IFSF score of diabetic women was 29.3 +/- 6.4 and was 37.7 +/- 3.5 in normal cases (p < 0.05). Lack of libido was the most common symptom in diabetics and was observed in 77% of the women. Diminished clitoral sensation was observed in 62.5% of the women, 37.5% complained of vaginal dryness and 41.6% had vaginal discomfort. Orgasmic dysfunction was found in 49% of the women. The incidence of all these related symptoms were significantly higher when compared to controls. We concluded that significant percentage of diabetic women that we observed experience sexual dysfunction of varying degrees that diminishes their quality of life. PMID- 11898711 TI - Magnetic resonance imaging (MRI) of sexual intercourse: second experience in missionary position and initial experience in posterior position. AB - Our objective was to confirm that it is feasible to take images of the male and female genitals during coitus and to compare this present study with previous theories and recent radiological studies of the anatomy during sexual intercourse. Magnetic resonance imaging was used to study the anatomy of the male and female genitals during coitus. Three experiments were performed with one couple in two positions and after male ejaculation. The images obtained confirmed that during intercourse in the missionary position, the penis reaches the anterior fornix with preferential contact of the anterior vaginal wall. The posterior bladder wall was pushed forward and upward and the uterus was pushed upward and backward. The images obtained from the rear-entry position showed for the first time that the penis seems to reach the posterior fornix with preferential contact of the posterior vaginal wall. In this position, the bladder and uterus were pushed forward. A different preferential contact of the penis with the female genitals was observed with each position. These images could contribute to a better understanding of the anatomy of sexual intercourse. PMID- 11898712 TI - Clinical trial development in female sexual dysfunction. AB - The development of clinical trial protocols for testing safety and efficacy of therapeutic interventions requires the blending of traditional methodologies with special consideration of the indications. The specificity of disease definition, precise identification of target populations, study design rationales, instruments, and selection of endpoints all contribute to the difficulty of conducting successful trials in this arena. An understanding of these issues can lead to improved clinical protocols. PMID- 11898713 TI - Unconsummated marriages: a separate and different clinical entity. AB - We bring our experience in the treatment of unconsummated marriages (UM). We postulate that it is an independent clinical entity. Since 1991, 199 couples with UM have sought treatment in our center in Buenos Aires. A new approach to this problem is based on an intensive treatment session that lasts a whole day and a pledge to pay only if therapy succeeds. The outcome was a success (97%) with coitus consummation. We make a long-term follow up. The advantages are the brevity of the treatment and lack of desertions. We postulate that UM cannot be approached with an individual perspective or with the usual psychotherapy techniques. PMID- 11898714 TI - [Pathogenesis of gallbladder cholesterolemia]. AB - Disturbances in cholesterol metabolism may be essential in pathogenesis of gallbladder cholesterosis (GBC). HDL cholesterol in the blood is subnormal. Physicochemical changes in the superficial layer of HDL induce impairment of free cholesterol esterification. Blood lipids and their apoprotein component are important for bile cholesterol level. In gallbladder contractile dysfunction but unaffected absorption there is enhanced passive and active cholesterol transport from the supersaturated bile to the cytoplasm of the epithelial cells from the bladder mucosa. Mechanism of intensive absorption of the lipids by macrophages operates primarily via modification of their apoprotein component. Modification of apoprotein occurs both in blood and gallbladder. Modified apoprotein is recognized by the macrophage and is absorbed by it together with transported lipids. In accumulation of great quantities of lipids in the macrophage it becomes big, slow, stays in the mucous or submucous layer of the wall and finally transforms into the foam cell. Moreover, deterioration of HDL cholesterol acception in GBC leads to slow discharge of cholesterol from the bladder wall. PMID- 11898715 TI - [Helicobacter pylori and ulcer dyspepsia]. PMID- 11898716 TI - [Echocardiographic alteration in patients with hemodynamic stroke]. AB - Echocardiography was made in 330 patients with ischemic stroke. Group 1 consisted of 72(21.8%) patients with hemodynamic stroke, the rest 258(78.2%) patients (group 2) had other pathogenetic subtypes of stroke. Chronic cardiac pathology was represented by postinfarction cardiosclerosis (38.9 and 17.8% for groups 1 and 2, respectively, p < 0.001) and continuous atrial fibrillation (15.3 and 15.1%, respectively, p > 0.05). Stroke volume of the left ventricle (LV), cardiac output and ejection fraction of the LV were significantly less than in group 2 and in control group but within normal range. The number of group 1 patients having local LV dyskinesia and transitory painless myocardial ischemia was significantly higher than in group 2 (p < 0.001). Painless ischemia correlated with local dyskinesia of the LV (p < 0.02). As transitory myocardial ischemia may deteriorate LV contractility and provoke excessive fall of blood pressure, local LV dyskinesia caused by postinfarction changes seems to be a principal and stable echocardiographic sign of developing hemodynamic stroke. PMID- 11898717 TI - [Neurological complications of infectious endocarditis]. AB - Of 204 patients with infectious endocarditis (IE) treated in the hospital in 1980 2000, 43(21.2%) developed neurological complications. These were: ischemic stroke (72.1%), hemorrhagic stroke (9.3%), both (7%), abscess and subarachnoidal hemorrhage (2.3% for each), meningitis (7%), toxic encephalopathy (11.6%). Neurological complications of IE arose prior to treatment and within the first week of antibacterial therapy in 63% cases, more frequently in the left carotid territory. Neurological complications in IE debute manifested acutely, pareses were more frequent than paralyses, with elevated temperature, low hemoglobin and red cell levels, leukocytosis. MRT detected 8 +/- 4.6 foci in the brain, CT--2 +/ 1.1, on the average. Lethality of IE patients with neurological complications reached 58.1% and was significantly higher than in those without such complications (14.9%, p < 0.001). Overall acturial survival 1 year after the discharge from the hospital was 94.4%, 5-year survival--61.1%, 10-year survival- 11%. PMID- 11898718 TI - [Clinical characteristics of endocarditis running with viral hepatitis in intravenous drug abusers]. AB - Viral hepatitis (VH) was diagnosed in 65% of patients with infectious endocarditis (IE) abusing intravenous narcotic drugs. VH caused recurrent course of IE in 8% examinees. The virusological tests of the blood for HBs-antigen or antibodies to viral hepatitis C were positive in 39 patients (group 1) and negative in 21 patients (group 2). VHB, VHC, VHB + VHC were diagnosed in 15, 57 and 28% patients, respectively. An acute course of IE was observed in 72% patients from group 1 and 76% patients from group 2, respectively, subacute course--in 20 and 24%, respectively, recurrent in 8% and 0%, respectively. IE patients with VH often had thrombohemorrhagic complications. DIC syndrome was detected in 25 and 4% patients, hemophthisis--in 33 and 28%, nephritis--in 71 and 48% patients, acute cerebral ischemia--in 7 and 4%, acute myocardial infarction- in 71 and 4%, hypocoagulation--in 53 and 8%, hypercoagulation--in 29 and 50% patients from group 1 and 2, respectively. PMID- 11898719 TI - [Thyroid hormones effects on hemodynamic and electrophysiological indices of heart performance and efficiency of antiarrhythmic drugs in paroxysms of cardiac fibrillation in patients with coronary heart disease]. AB - The article provides the results of thyrotropic hormones (TH) influence on spontaneous paroxysms of cardiac fibrillation (CFP), central hemodynamics, electrophysiology (EP) of the heart and effectiveness of antiarrhythmic drugs (AAD) in patients with coronary heart disease (CHD). It is shown that in CFP patients TH should be evaluated in sinus rhythm and in arrhythmia. Fluctuations in TH contribute to CFP. EP and hemodynamic effects of TH manifest with shorter effective refractory period of the left atrium and atrioventricular junction, disturbed diastolic function of the left ventricle. TH affect efficiency of AAD in CFP patients with CHD. PMID- 11898720 TI - [The role of molecular genetic diagnosis in prediction and prevention of age specific pathology]. AB - A method of molecular-genetic diagnosis of diseases in hereditary predisposition developed by the authors is novel in geriatric practice. It allows to predict such prevalent age-specific diseases as ischemic heart disease, hypertension, noninsulin-dependent diabetes mellitus, Alzheimer's and some malignancies. Assessment of molecular-genetic, clinical and laboratory data can identify predisposition or early symptoms of the disease. Special system of prophylaxis and pathogenetic treatment is designed to prevent development of genetically predetermined age-specific diseases. PMID- 11898721 TI - [Influence polyunsaturated fatty acids omega-3 on coagulation and fibrinolysis systems in patients with NIDDM]. AB - 35 patients with diabetes mellitus type 2 complicated with hyperlipidemia, coronary heart disease, were treated for 8 months with poseidonol which incorporates essential polyunsaturated fatty acids and fat-soluble vitamins. Poseidonol reduced atherogenic index from 4.2 to 3.15. The study of blood coagulation and fibrinolysis points to signs of hypocoagulation and normalization of fibrinolytic activity against previous hypercoagulation and fibrinolysis depression. PMID- 11898722 TI - [Effects of aspirin standard therapy and chronotherapy on circadian organization of hemocoagulation in patients with insulin-dependent diabetes mellitus]. AB - 24-h profile of hemocoagulation was assessed in 30 patients with insulin dependent diabetes mellitus type 1 aged 17-37 years. The patients were randomized into 2 groups, 15 patients each. Patients of group 1 received aspirin by conventional scheme: 125 mg 3 times a day for 16 days. Patients of group 2 received aspirin as preventive chronotherapy: once a day in a dose 125 mg for 16 days two hours before the acrophase of platelet aggregation rhythm--at 22 p.m. Parameters of plasmic and platelet hemostasis in blood samples were measured at 3.00, 7.00, 11.00 a.m., 15.00, 19.00, 23.00. The above chronobiological information was processed by Kosinor-analysis according to F. Halberg. Before the treatment, hypercoagulation with night rise as well as external and internal desynchronism of the hemostasis circadian rhythms were observed. Conventional aspirin treatment improved hemostasis but influenced acrophases minimally. Aspirin chronotherapy promoted normalization of circadian organization of hemocoagulation. PMID- 11898723 TI - [Clinical and diagnostic aspects of drug-induced hepatitis]. AB - Drug-induced hepatitides (DIH) may manifest with a wide range of symptoms from insignificant activation of the enzymes with asthenic syndrome to severe disease with jaundice. The most informative tests for early diagnosis of medicinal lesion of the liver are biochemical tests of blood serum with determination of activity of the enzymes AlAt, AsAT, AP, GGTP. Long-term use of the drug and late diagnosis of DIH endanger drug damage to the liver. Treatment of acute drug hepatitides consists in administration of medication stimulating metabolic ability of hepatic cells. Acute DIH is characterized by the absence of splenomegaly. PMID- 11898724 TI - [Outpatient treatment of irritable colon syndrome: quality of diagnosis and treatment]. AB - 40 patients with irritable colon syndrome were treated outpatiently: 26 patients by gastroenterologist and 14 ones by therapist. Management of such patients by gastroenterologist significantly lowered frequency of their hospitalizations and sick leave duration as compared to those treated by therapist. PMID- 11898725 TI - [Circadian monitoring of intragastric pH in evaluation of efficiency of antisecretory therapy of duodenal ulcer]. AB - Inhibitors of proton pump are thought "golden standard" in the treatment of ulcer. Usage of omeprasol generics in standard doses sometimes fail to adequately reduce acid production. In pain persistence in patients with duodenal ulcer receiving standard antisecretory therapy it is effective to control it with 24-h intragastric pH-metry which either indicate the necessity of dose correction or another cause of pain (concomitant affections as a rule). As shown by 24-h pH metry, a new generic of omeprasol, omeprus, is close by effectiveness to the known drug omeaz. PMID- 11898726 TI - [Efficiency of antiarrhythmic drugs of class 1C and isoptin in paroxysmal supraventricular tachycardia]. AB - Cross-over trial studied comparative effectiveness and tolerance of allapinine, rhythmonorm and isoptine in patients with paroxysmal supraventricular tachycardia (PSVT). Treatment effects did not differ much. Analysis of the preventive efficiency of the above drugs with reference to PSVT form has demonstrated that in paroxysmal atrioventricular reciprocal tachycardia the highest activity was shown by isoptine and rhythmonorm, in paroxysmal orthodromic reciprocal tachycardia--by rhythmonorm and allapinin. In long-term therapy, a preventive effects of allapinin, rhythmonorm and isoptine achieved at short-term course therapy persisted in 90.9, 95.6 and 81.5% patients, respectively. All the above drugs cause moderate inhibition of atrioventricular and intraventricular conduction. Isoptin had the best tolerance. PMID- 11898727 TI - [Effectiveness of sparfloxacin (sparflo) in the treatment of complicated forms of pyelonephritis and prostatitis]. AB - A novel difluoroquinolone drug sparfloxacine has a wide antibacterial spectrum. It is active both against gram-negative and gram-positive flora. Sparfloxacine was tried in 43 patients with pyelonephritis and prostatitis complicated with urolithiasis, diabetes mellitus, nephroptosis, anomalous kidneys, etc. The drug was given according to its pharmacokinetics for 7-14 days in a daily dose 400 mg (day 1) then 200 mg/day. The effectiveness of sparfloxacine in complicated pyelonephritis and prostatitis was rather high: clinical response reached 83.7%, bacteriological one--78.7%. PMID- 11898728 TI - [Experience with nebulizer therapy of severe bronchial asthma]. PMID- 11898730 TI - [Information on All-Russia Symposium "Long Q-T Interval Syndrome". Moscow, March 22-23, 2001]. PMID- 11898729 TI - [Problems of congestive heart failure at the XXII Congress of European Cardiology Society (Amsterdam, Netherlands, August 2000). Part 2]. PMID- 11898731 TI - [Gout: current views. Stage oriented treatment]. AB - Gout is a common disease arising due to abnormal purin metabolism and excessive accumulation of uric acid in the blood (hyperuricemia) and manifesting with attacks of acute gouty arthritis. In long duration of gout uric acids accumulate in the bones and periarticular tissues as tophuses. Repeat attacks lead to development of chronic gouty arthritis. Purins restriction diet is an important component of gout treatment. Treatment of acute arthritis should be started early, in initial pains before the development of the attacks. Gouty arthritis in the presence of continuous hyperuricemia, tophyses and urolithiasis is treated with allopurinol. Its intake should be long and controlled by the blood level of uric acid. Balneotherapy is recommended for patients with chronic gouty arthritis associated with cardiovascular diseases, urolithiasis. PMID- 11898732 TI - [Role of alcohol in heroin overdose]. AB - The relationship between ethanol and risk of heroin overdosage was studied. Statistical processing of the results of forensic chemical analysis (460 expert evaluations) carried out in Chelyabinsk Regional Bureau of Forensic Medical Expert Evaluations in 2000 was carried out. The results of morphine and ethanol measurements in the blood and urine from corpses where deaths ensued from narcotic or ethanol poisoning, were analyzed. The concentrations of morphine in the blood and urine were measured on a gaseous chromatographer with mass selective detector (Hewlett Packard HP 6890/HP-5972). Methods for measuring urinary and blood morphine are described. The results of statistical analysis demonstrated relationships between the age and ethanol concentrations in the blood and urine; blood ethanol and total urinary and blood morphine concentrations; blood concentration of free morphine and presence of 6 monoacetylmorphine in the blood. The authors conclude that the presence of ethanol in the blood together with morphine drastically augments the risk of rapid death from respiration arrest. It can also lead to a relatively high risk of overdosage in experienced narcomaniacs using heroin and ethanol. PMID- 11898733 TI - [Forensic medical significance of enzymatic products of ethanol oxidation in cadaveric brain]. AB - The concentrations of ethanol, acetaldehyde, and the oxidizing enzymes alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (AIDH) were measured in neuronal cytoplasm, limbic cortical capillaries, and cardiovascular center of the medulla oblongata. The measurements were carried out by histochemical methods, gas-liquid chromatography, etc. The results were processed with consideration for the degree and stage of ethanol intoxication in case of death from ethanol poisoning and asphyxia in hanging. Increase of ethanol concentration in the blood was associated with a decrease and then increase in the brain concentrations of ADH and with an increase of AIDH concentration. Enzymatic changes predominated in capillary walls; the minimum shifts were observed in the neuronal cytoplasm of the cerebral limbic cortex, which confirms the neurohumoral nature of detoxication regulation. Lethal ethanol poisoning could occur during any stage of ethanol intoxication. The detected changes in ethanol, acetaldehyde, and metabolizing oxidoreductases in brain tissue can be used for forensic medical diagnosis of ethanol poisoning. PMID- 11898734 TI - [Use of liver morphometry in the differential diagnosis of chronic alcohol and narcotic poisoning]. AB - Data on the quantitative histomorphological involvement of the liver in chronic narcotic and ethanol poisoning and their combination are presented. Alcohol use by narcomaniacs augments hepatocyte damage, which manifests by increased fatty degeneration, increased activity of lobular hepatitis, appearance of the neutrophil admixture in inflammatory infiltration, and stimulation of sclerotic processes leading to an increase in the perimeter of portal tract section at the expense of connective tissue growth. Quantitative analysis of these processes is recommended for forensic medical histological differential diagnosis of chronic narcotic and ethanol poisoning. PMID- 11898735 TI - [On racial-ethnic features of the mucosal structure of the gingival edge and tongue]. AB - The structure of the mucosal membrane of the gingiva and dorsal surface of the tongue was studied in 415 residents (men and women aged 18-35 years) of Africa, Southern, Central, and Northern Asia, and Russia in order to detect racial and ethnic differences. A complex of modern methods of investigation was used (use of intraoral videotape recorder, digital photo, computer analysis, plaster models, uni- and multi-dimensional mathematical analysis). The data can be practically important in forensic medical personality identification. PMID- 11898736 TI - [Preservation of methanol in cadaveric material]. AB - Duration of preservation of methanol in cadaveric material at 20 +/- 3 degrees C during 4-13 months was studied on 19 model and expert objects of cadaveric material (muscle tissue, kidney, blood, urine). Exponential relationship between methanol concentration and duration of storage of the object was detected. Methanol concentration decreased by 30-70% during the first weeks, after which the process decelerated to 5-10% a month. Toxicologically significant concentrations of endogenous methanol were not revealed in any case. PMID- 11898738 TI - [Crystallization of cerebrospinal fluid in cases of death from ischemic heart disease and ethanol poisoning]. AB - Crystallographic analysis of the cerebrospinal fluid (CSF) was carried out in 18 cases of death from coronary disease and 19 cases of death from ethanol poisoning. The crystallograms were evaluated visually and by the stereoscopic picture. Specific features of the CSF crystal colonies growth in subjects dead from the above conditions are determined. PMID- 11898737 TI - [Comparative characteristics of various methods of sample preparation for determining ethanol in muscle tissue]. AB - Three methods for sample preparation for ethanol detection by gas chromatography are compared. The highest ethanol output was obtained after preparation by 5-min heating of the biological object and the internal reference object with addition of trichloroacetic acid as the acidifier. Relationship between the results of ethanol measurements and the degree of muscle crushing was evaluated. PMID- 11898739 TI - [Use of the portable hymenoscope PHS-1 for genital examination in cases of rape]. PMID- 11898740 TI - [Analysis of synthetic pyrethroids by gas chromatography-mass spectrometry]. PMID- 11898741 TI - [Radiation loading, radiation-induced diseases and suicide values in a region adjacent to a nuclear testing ground]. AB - Radiation effects on the population health at a territory adjacent to the Semipalatinsk nuclear testing grounds were studied. Immunosuppression and suppression of nonspecific defense factors, induced by radiation from the testing grounds, determine the extension of physiological lability range, accelerate the natural involution processes, and create psychoemotional stress. The totality of radiation-induced general physiological effects with pronounced dose dependence of their incidence and intensity represents a co-factor of suicide. PMID- 11898742 TI - [P.A. Aliavdin, graduate of Moscow University was the organizer of the Forensic Medical Corp of the Ivanovskiy district]. PMID- 11898743 TI - [International conference "Forensic-Dental Expertise: status, prospects for development and improvement (24-25 May 2001)]. PMID- 11898744 TI - [Deposition and incorporation of trace objects on the bone in blunt injury]. AB - A total of 457 cases with the impact of blunt hard objects and cars on the surface of different bones with formation of sites of deposition and incorporation of trace objects are analyzed. Variants of incorporation of fragments of hairs, fibers, metals, varnish into bone surface and microstructure are described, their diagnostic significance, mechanism of formation, and relationship with bone injuries are discussed. PMID- 11898745 TI - Getting paid: wrestling with the system. PMID- 11898746 TI - Majority of visits equal 99214. PMID- 11898747 TI - HIPAA and mental health notes. PMID- 11898749 TI - Six health plans to begin paying for quality. PMID- 11898748 TI - Seven tips to improve your ICD-9 coding for diagnostic tests. PMID- 11898750 TI - Going solo: making the leap. AB - Frustrated by current practice and convinced of a better way, the author left his salaried position and opened a solo practice with no staff. Because his overhead costs are extremely low, the author is able to see fewer patients per day and create more meaningful interactions. By offering unfettered access, the author finds that his patients trust him more and actually call him less. PMID- 11898751 TI - Improving anticoagulation management at the point of care. AB - Many physicians and other providers attempt therapeutic warfarin oversight without regularly scheduled anticoagulation appointments. Studies show that the risk of major bleeding or thromboembolic events due to warfarin therapy is between 2 percent and 12 percent per year. Point-of-care anticoagulation devices are convenient for patients and physicians and allow for patient-focused anticoagulation care. PMID- 11898752 TI - Test your coding skills. PMID- 11898753 TI - Results from the FPM practice self-test. PMID- 11898754 TI - Horses vs. zebras II. PMID- 11898755 TI - It's not too early to prepare for semi-retirement. PMID- 11898756 TI - Precepting: help yourself while helping students. PMID- 11898757 TI - Reflections on the phenomenon of readiness for death. AB - My recent study investigated the phenomenon of readiness for death by analysing texts resulting from interviews with ten participants confronting their own death. The methodology was grounded in phenomenological philosophy. This paper focuses on the interpretations of the experiences of one participant, Charlie. Individual themes are discussed in the context of broader (common) themes to highlight Charlie's readiness for death. PMID- 11898758 TI - Dying in the curative system: the haematology/oncology dilemma. Part 1. AB - Findings from qualitative research exploring the experience of a group of carers of patients with haematological malignancies in relation to end-of-life are presented as a contribution to documenting the emergent issues in this area. The findings are set in the context of parallel research on a group of carers who have experienced hospice care during the dying trajectory of their loved one. It is shown by comparison that the institutional deaths of this group of patients in haematology/oncology was far from the best practice end-of-life care that the hospice group received. PMID- 11898759 TI - Cross-cultural communication and health care practice. AB - This article describes concepts such as culture, cross-culture and the effect of culture in the communication process between clients and health professionals from culturally and linguistically diverse backgrounds in order to promote holistic health care. The paper outlines a case study that can be used to both support and improve cultural competence. PMID- 11898760 TI - Nutrition education and promotion in primary schools. AB - Health promotion with school-age children can enhance knowledge about nutrition and change eating habits when health workers, schools, and communities work together for a common goal. A range of options need to be considered when designing an effective and appropriate school-based program. PMID- 11898761 TI - The domination of chronic illness research by biomedical interests. AB - The interest of biomedicine is the physical body viewed in isolation from the contextual understandings which shape the chronic illness experience. This neglect of the illness experience was evident in an inquiry conducted by the first author with 81 women who live with chronic illness. Herein we discuss a secondary analysis of the correspondence data in which we found many common elements across different medical diagnoses. PMID- 11898762 TI - [Irina Iakovlevna Zakharova - 75th Birthday Jubilee]. PMID- 11898763 TI - Francis Trevelyan Buckland (1826-1880). PMID- 11898764 TI - William Marsden (1796-1867); Alexander Marsden (1832-1902). PMID- 11898765 TI - Conflict of interest and the credibility of nicotine and tobacco research. PMID- 11898766 TI - Social Anxiety: From Laboratory Studies to Clinical Practice. Symposium proceedings. March 22, 2001. PMID- 11898767 TI - Comparison of LDL fatty acid and carotenoid concentrations and oxidative resistance of LDL in volunteers from countries with different rates of cardiovascular disease. AB - Within Europe there are differences in cardiovascular disease (CVD) risk between countries and this might be related to dietary habits. Oxidative modification of LDL is suggested to increase the risk of CVD and both the fatty acid and antioxidant content of LDL can affect its oxidation. In the present study, concentration of LDL fatty acid and antioxidant micronutrients (tocopherols and carotenoids) and ex vivo oxidative resistance of LDL (lag phase) was compared in volunteers from five countries with different fruit and vegetable intakes and reported rates of CVD. Eighty volunteers (forty males, forty females per centre), age range 25-45 years, were recruited from France, Northern Ireland, UK, Republic of Ireland, The Netherlands, and Spain, and their LDL composition and lag phase were measured. There were some differences in LDL carotenoid and alpha-tocopherol concentrations between countries. alpha-Tocopherol was low and beta- + gamma tocopherol were high (P<0.001) in the Dutch subjects. Beta-Carotene concentrations were significantly different between the French and Spanish volunteers, with French showing the highest and Spanish the lowest concentration. LDL lycopene was not different between centres in contrast to lutein, which was highest in French (twofold that in the Dutch and Spanish and threefold that in Northern Ireland and the Republic of Ireland, P<0.001). However absolute LDL saturated, monounsaturated, polyunsaturated and total unsaturated fatty acid concentrations were different between countries (P<0.001, total unsaturated highest in Northern Ireland) there was little difference in unsaturated:saturated fatty acid concentration ratios and no difference in polyunsaturated:saturated fatty acid concentration ratios. LDL from the Republic of Ireland (a region with a high rate of CVD) had greater resistance to Cu-stimulated oxidation than samples obtained from volunteers in other countries. In conclusion, LDL composition did not predict resistance to Cu-stimulated oxidation, nor is there evidence that LDL from volunteers in countries with lower rates of CVD have greater resistance to oxidation. PMID- 11898768 TI - Antioxidant vitamin supplements do not reduce reactive oxygen species activity in Helicobacter pylori gastritis in the short term. AB - Reactive oxygen species have been implicated in Helicobacter pylori-mediated gastric carcinogenesis, whereas diets high in antioxidant vitamins C and E are protective. We have examined the effect of vitamin C and E supplements in combination with H. pylori eradication on reactive oxygen species activity in H. pylori gastritis. H. pylori-positive patients were randomized into four groups: triple therapy alone (Bismuth chelate, tetracycline, and metronidazole for 2 weeks), vitamins alone (200mg vitamin C and 50mg vitamin E, both twice per day for 4 weeks), both treatments or neither. Plasma and mucosal ascorbic acid, malondialdehyde and reactive oxygen species were determined before and after treatment. Compared with normal controls (n 61), H. pylori-positive patients (n 117) had higher mucosal reactive oxygen species and malondialdehyde levels and lower plasma ascorbic acid. Plasma ascorbic acid doubled in both groups of patients receiving vitamins and mucosal levels also increased. Malondialdehyde and reactive oxygen species fell in patients in whom H. pylori was eradicated but vitamin supplements were not effective either alone or in combination with H. pylori eradication. Supplements of vitamins C and E do not significantly reduce mucosal reactive oxygen species damage in H. pylori gastritis. PMID- 11898769 TI - Associations between spontaneous meal initiations and blood glucose dynamics in overweight men in negative energy balance. AB - The aim of the present study was to investigate associations between spontaneous meal initiations and blood glucose dynamics in overweight male subjects in negative energy balance. In a randomized crossover design, fifteen overweight male subjects (BMI 28.6 (SD 1.8 kg/m2) participated in three treatments, each of which consisted of 2 weeks consuming a low-energy diet followed by a test of voluntary food ingestion in the absence of time-related cues. The low-energy diet consisted of three daily meals (947 kJ) which were either semi-solid with or without 2.5 g guar gum, or solid, and a dinner of subject's own choice. During the time-blinded test, on the first, second, and third meal initiation subjects ingested a low-energy meal corresponding to that used during the preceding weeks. Changes in blood glucose were monitored on-line. Associations between spontaneous meal initiations and blood glucose dynamics were determined using the chi2 test. No difference was found between treatments in the occurrence of postabsorptive and postprandial declines in blood glucose or in associations between meal initiations and blood glucose dynamics. Postprandial dynamic blood glucose declines were associated with meal initiation (chi2 26 8, P<0.00 1), but postabsorptive and postprandial transient declines were not. In overweight subjects, the usual association between transient declines and spontaneous meal initiation was completely absent in negative energy balance. PMID- 11898771 TI - Science trumps politics. PMID- 11898770 TI - Total antioxidant and ascorbic acid content of fresh fruits and vegetables: implications for dietary planning and food preservation. AB - Epidemiological evidence links high intake of ascorbic acid (AA) and other antioxidant micronutrients to health promotion. It would be useful to know the overall, or 'total' antioxidant capacity of foods, to establish the contribution of AA to this, and to assess how this information may translate into dietary intakes to meet the new US daily reference intake for AA. In this study, the total antioxidant capacity, as the ferric reducing-antioxidant power (FRAP) value, and AA content of thirty-four types of fruits and vegetables were measured using a modified version of the FRAP assay, known as FRASC. This measures AA (reduced form only) simultaneously with the FRAP value. Results covered a wide range: 880-15940 micromol/kg fresh wet weight and <20-540 mg/kg fresh wet weight respectively, for FRAP and AA, which comprised < 1-73 % and < 1-59 % total antioxidant capacity of fruits and vegetables respectively. We estimate that 100 mg AA is contained in one orange, a few strawberries, one kiwi fruit, 1-2 slices of pineapple, several florets of raw cauliflower or a handful of uncooked spinach leaves. Apples, bananas, pears and plums, the most commonly consumed fruits in the UK, contain very little AA. Results indicate also that the antioxidant capacity of vegetables decreases rapidly and significantly after fragmentation. Results of this, and future studies, using FRASC as a biomonitoring tool will be useful in food production, preparation, preservation, and aid dietary choices to increase antioxidant and AA intake. Furthermore, FRASC will facilitate bioavailability studies of antioxidants from different foods of known antioxidant capacity and AA content. PMID- 11898772 TI - Are the effects of dietary fruits and vegetables on human health related to those of chronic dietary restriction on animal longevity and disease? PMID- 11898773 TI - Parenteral lipid emulsions and phagocytic systems. AB - Lipid emulsions (LE) for parenteral use are complex emulsions containing fatty acids, glycerol, phospholipids and tocopherol in variable amounts and concentrations. In clinical practice, LE have been employed for more than 30 years. Fatty acids may have different impacts on phagocytic cells according to their structure. Experimental and clinical studies have consistently shown that LE modify monocyte/macrophage and polymorphonuclear phagocytosis. The inhibitory effect of LE on the functional activity of the phagocytic system, although still clinically controversial, may have a harmful impact because total parenteral nutrition with lipids may be recommended in hypercatabolic conditions where inflammation and infection are present. LE based on triglycerides containing long chain fatty acids (termed long chain triglycerides or LCT) are the main parenteral fat source and are typically rich in n-6 polyunsaturated fatty acids. They may have adverse effects on the immune system, especially when given in high doses over a short period of time. However when administered properly they can be used safely. LE containing medium chain triglycerides (MCT) may have some advantages because of their positive effects on polymorphonuclear cells, macrophages, and cytokine production, particularly in critically ill or immunocompromised patients. New parenteral LE containing n-3 polyunsaturated fatty acids or monounsaturated olive oil are already available in Europe. Judicious use of these new LE is mandatory especially relating on their potential impact on the immune system. New experimental and clinical studies are required to further establish the role of LE in clinical nutrition. PMID- 11898774 TI - Impact of parenteral n-3 fatty acids on experimental acute colitis. AB - The present study was undertaken to investigate the effects of parenteral lipid emulsions (LE) enriched with n-3 fatty acids (n-3 FA) in experimental acute colitis. Seventy-four adult male Wistar rats were randomized into six groups, five of which had acetic acid-induced colitis. The animals received a fat-free diet and water ad libitum in individual metabolic cages. By a central venous catheter, saline was infused (0.5 ml/h) into the control groups CS (without colitis) and CC (with colitis), while the test groups received specific LE for 7 days. The n-3/n-6 FA ratio and the lipidic compositions regarding long chain (LCT) and medium chain (MCT) triglycerides were: group L--1:7.7 (LCT, n = 12), M- 1:7.0 (MCT and LCT, n = 12), LW-3--1:4.5 (LCT plus n-3 FA, n = 12) and MW-3- 1:3.0 (MCT and LCT plus n-3 FA, n = 13). The frequency of diarrhea, oral intake/body weight ratio, intestinal alterations, macrophage cellularity were evaluated and colonic concentrations of leukotrienes (LTB4, LTC4), prostaglandins (PGE2) and thromboxanes (TXB2) were measured. Groups M, MW-3 and LW-3 had less diarrhea than the CC group (P<0.05). Average oral intake/body weight ratio in MW 3 animals was comparable to the CS and better than the CC group. n-3 FA treated rats (LW-3 and MW-3) presented less intestinal inflammatory alterations than CC rats. Mucosal ulcer formation in MW-3 group did not differ from CS rats. M and MW 3 rats had less macrophages in the colon than the CC group. Compared with CC group, lower concentrations of LTB4 in the CS, LW-3 and MW-3 groups; of PGE2 in the CS, M and MW-3 groups; and of TXB2 in the CS and MW-3 groups were found. Mean concentrations of LTC4 did not differ among the groups. Thus, a LCT-containing LE with a low n-3-n-6 ratio does not modify inflammatory colitis manifestations; LE with a high n-3-n-6 ratio reduces diarrhea, preserves oral intake-weight ratio, attenuates morphological consequences and decreases colonic concentrations of inflammatory mediators; MCT/LCT-containing LE with 1:3 n-3-n-6 ratio exerts the most profound beneficial impact on the inflammatory response. PMID- 11898775 TI - Top type Is for accredited organizations. Track your performance in these trouble spots. PMID- 11898776 TI - Codman Award winners describe improvements. Recognition for excellence in use of outcomes measurement to achieve improvement. PMID- 11898777 TI - New and revised credentialing and privileging standards for long term care and subacute programs. PMID- 11898778 TI - [Grain, farmer and persistent eye inflammation]. PMID- 11898779 TI - Re: neurophysiology not required before surgery for typical carpal tunnel syndrome. PMID- 11898780 TI - Royal College of Radiologists Annual Undergraduate Essay Prize. Melanoma: the new smallpox? Can vaccines be used to treat melanoma? AB - This essay assesses the effectiveness of vaccine therapy for melanoma. Risks and benefits of various vaccine strategies are explored, as are the processes by which such therapies are assessed. An overview of cancer immunobiology underlying vaccine therapy is given. PMID- 11898781 TI - Radiosurgery for brain metastases. PMID- 11898782 TI - Cancer gene therapy: Part 1. Vector development and regulation of gene expression. PMID- 11898783 TI - Accuracy of a relocatable stereotactic radiotherapy head frame evaluated by use of a depth helmet. AB - In high precision radiotherapy, the more accurately the patient can be relocated, the smaller the clinical to planning target volume margin can be, with reduction in the volume of normal tissue irradiated. The Gill-Thomas-Cosman (GTC) relocatable stereotactic head frame provides immobilization of the patient which is highly reproducible. A depth helmet and measuring probe were used to confirm the accuracy of relocation of 31 patients treated in the GTC frame. The measurements were processed in a spreadsheet developed to calculate the size of the patient's displacement as a vector. Twenty-seven patients received fractionated stereotactically-guided conformal radiotherapy, and 4 single fraction stereotactic radiosurgery, amounting to 564 measurement episodes. The accuracy was extremely good, and considerably more accurate than standard thermoplastic head shells. Ninety-two percent of the displacement vectors were less than 2 mm, and 97% less than 2.5 mm. Considering each dimension separately, the largest mean displacement was 0.4 mm in the superior-inferior direction. Accuracy was constant through a fractionated course for most patients, but prediction based on measurements from the first few fractions was not reliable. Results were dependent on patient selection, with worse reproducibility in patients with neurological deficits, or difficulty cooperating. The depth helmet measurements detected a loosened mouth bite in one patient and allowed repositioning to be verified without the need for the simulator. Total treatment time, including use of the depth helmet to verify treatment position, is quicker (mean 15.7 min) than using portal films. The depth helmet, used in conjunction with the vector displacement spreadsheet, provides a simple way to define the CTV PTV margin. For fractionated stereotactic radiotherapy we use a 3 mm CTV-PTV margin. This system could assist technology transfer to centres starting stereotactic radiotherapy using the GTC frame. PMID- 11898784 TI - Treatment-related morbidity and hospital league tables: experience from a national audit of radiotherapy-induced morbidity in cervical carcinoma. AB - Data are now available from a U.K. audit of survival and late morbidity following curative radiotherapy for cancer of the cervix treated in 1993. The complication rate per centre ranges from 0 to 67%. Although the frequency of complications following curative radiotherapy for cancer of the cervix might be considered to be an indicator of clinical performance, variation in treatment outcomes can be explained by sampling variability rather than real differences in quality of care. In the present study we have asked the question: could the disparity in complication rates between centres be no more or less than would be expected by chance? Our analysis suggests that this is the case, and for this reason it would be premature to use such outcome data to produce league tables or to assess institutional differences. Thus, ranking centres according to complication rate would not be valid, as the differences in rates observed are probably not significantly different from the national average. It is important that audit data are not used inappropriately and this analysis further highlights the need for reliable prospective collection of clinical information and the importance of considering sampling variability in interpreting the results of such studies. PMID- 11898785 TI - Radiation therapy for cervix carcinoma: benefits of individualized dosimetry. AB - This study is a presentation of the prospective collection of data on patients treated by radical radiotherapy at Westmead Hospital between December 1989 and December 1998. The impact of the routine use of individualized dosimetry and lower brachytherapy dose on patients was examined, by comparing the historical series of patients treated between 1989 to 1991 to the later patients treated with inividualised dosimetry. There were 163 patients treated with external beam and intracavitary radiotherapy during this period. Histology was squamous carcinoma in 80% (132 patients), adenocarcinoma in 13% (22 patients), and adenosquamous carcinoma in 6% (9 patients). Patients were generally treated with 50 Gy in 25 fractions to the pelvis followed by 1 low dose rate caesium intracavitary brachytherapy insertion. Patients who had dosimetry generally received 20 Gy to point A via the insertion compared to 30 Gy in the non dosimetry group. Median follow-up was 62 months. Only 22% (18) of patients failed with disease outside the pelvis. Pelvic control was similar in the patients who had dosimetry as opposed to no dosimetry (P=0.8). In the dosimetry group there were less grade III or higher bowel toxicity (P=0.01) and less vaginal fistulae (P=0.03). The actuarial two-year survival was 56.2% in the no dosimetry group and 68.6% in the dosimetry group. When controlled for stage and performance status patients who had dosimetry had a statistically significant greater overall survival (P=0.02). Thus we found that the routine use of dosimetry was associated with a lower brachytherapy dose, decreased complications, without any decrease in local control or survival. PMID- 11898786 TI - Uterine sarcomas--the biggest challenge? PMID- 11898787 TI - Locoregional treatment in breast cancer: new paradigm raises new questions. PMID- 11898788 TI - Weight gain during adjuvant and neoadjuvant chemotherapy for breast cancer: an audit of 100 women receiving FEC or CMF chemotherapy. AB - Weight gain is reported as a common finding in patients treated for breast cancer but its aetiology appears to be complex. The aim of this study was to investigate the incidence and degree of weight gain during chemotherapy and to examine possible contributory factors. Data were collected on 100 women treated with CMF or FEC chemotherapy. The mean change in weight was +3.68 kg (P<0.001). 64% of patients gained more than 2 kg in weight, 31% maintained a stable weight (within + or - 2 kg) and 5 patients lost more than 2 kg. Approximately 1/3 of patients (33) gained more than 5 kg and 6 patients gained more than 10 kg in weight. The majority of patients (85%) received steroids as antiemetics but no effect of steroid dose was seen on the level of weight change. No significant differences in weight gain were seen in patients receiving tamoxifen (37%) compared with those not taking it. Similarly, menopausal status did not appear to be a significant factor influencing weight gain. In summary, a high incidence of weight gain was found. The literature on weight gain in breast cancer and possible interventions to avoid weight gain are discussed. PMID- 11898789 TI - Women and smoking: time for a change. PMID- 11898790 TI - Meningeal carcinomatosis in rectal cancer. PMID- 11898791 TI - Training in clinical oncology--how much is enough palliative medicine? PMID- 11898792 TI - Trainee in clinical oncology--how much is enough palliative medicine? PMID- 11898793 TI - Adrenal cortical carcinoma presenting initially with radius metastasis. PMID- 11898794 TI - A test of whether selection maintains isochores using sites polymorphic for Alu and L1 element insertions. PMID- 11898795 TI - Modeling thin filament cooperativity. PMID- 11898796 TI - On the potential functions used in molecular dynamics simulations of ion channels. PMID- 11898797 TI - Nucleus basalis of Meynert pathology in the human brain after fatal head injury. AB - Dysfunction of the basal forebrain cholinergic system has been hypothesized to contribute to deficits of memory and cognition after head injury. We have previously reported reduced levels of choline acetyltransferase activity in the cerebral cortex of patients who died after a head injury, demonstrating that there is a loss of cortical cholinergic innervation. In the present study, we examined the nucleus basalis of Meynert (NBM), which provides cortical cholinergic innervation, in fatally head-injured patients and in controls. The majority of head-injured patients had histological evidence of neuronal damage in the NBM, which was due to mechanical distortion of the tissue and/or ischemic damage. The findings demonstrate that the NBM is vulnerable after severe head injury and that secondary insults play an important role in the damage. Thus, both neuronal perikarya and terminals of the basal forebrain cholinergic system are damaged after human fatal head injury. This damage may contribute to persisting dysfunction of memory and cognition in head-injured patients who survive. PMID- 11898798 TI - Cases from the Osler Medical Service at Johns Hopkins University. B(12) deficiency. PMID- 11898799 TI - Assessment of risks of brodifacoum to non-target birds and mammals in New Zealand. AB - The risks to non-target birds and other wildlife from the use of vertebrate pesticides, including anticoagulant rodenticides, are determined to a significant extent by species' intrinsic susceptibility, and the toxicokinetics of the compounds used. Brodifacoum is highly toxic to birds and mammals. The acute toxicity of brodifacoum to birds in New Zealand varies from <1 mg/kg in pukeko (Porphyrio p. melanotus), the native swamp hen, to >20 mg/kg in the paradise shelduck (Tadorna variegata). Like other second-generation anticoagulants brodifacoum is strongly bound to vitamin K epoxide reductase and will persist, apparently for at least 6 months, in organs and tissue containing this enzyme, e.g., liver, kidney, and pancreas. The unique toxicokinetics of this class of compound exacerbates the risk of primary and secondary poisoning of non-target species. Vertebrate pest control programmes in New Zealand using bait containing brodifacoum have resulted in the primary and secondary poisoning and sub-lethal contamination of non-target species. These include native raptors, such as the Australasian harrier (Circus approximans) and morepork (Ninox novaeseelandiae), other native birds such as the pukeko, weka (Gallirallus australis), southern black-backed gull (Larus dominicanus), and kiwi (Apteryx spp.), and introduced mammals, including game animals. There are increasing numbers of reports worldwide of wildlife contamination and toxicosis after the use of second generation anticoagulants. All pest control activities require careful risk benefit assessment in view of their potential to cause adverse environmental impact. Monitoring of wildlife for pesticide residues will provide data that can be used to reduce the risk of anticoagulant bioaccumulation and mortality in non target species. PMID- 11898800 TI - Risk to breeding success of Ardeids by contaminants in Hong Kong: evidence from trace metals in feathers. AB - The feathers of two Ardeid species, the Little Egret (Egretta garzetta) and the Black-crowned Night Heron (Nycticorax nycticorax) were collected from six egretries and two egretries respectively, located in different areas in the New Territories of Hong Kong, including the Mai Po Marshes (within a Ramsar site). These feathers were digested and concentrations (microg/g dry weight) of copper (4.6-19.4), iron (8.1-641.3), manganese (0.4-19.4), zinc (51.3-183.5), lead (0.1 5.1), cadmium (0.01-0.15), chromium (0.06-1.7) and mercury (0.0-7.1) were determined by ICP-AES, ICP-MS and CVAAS. The levels of manganese, mercury and lead found were equal to or less than the concentrations found in previous investigations, reflecting a slight downward trend most apparent with lead. As a general rule, the levels of lead and mercury were higher in the egretries close to the polluted Deep Bay. A probabilistic risk assessment of the possible adverse effects on the breeding success of the Little Egret was carried out with respect to mercury, lead and cadmium. It was concluded that mercury (0.5-7.1 microg/g dry weight feathers) probably has had adverse effects at the Au Tau egretry of the Little Egrets, but there was no evidence of adverse effects at other egretries. The probabilistic analysis also indicated a low likelihood of adverse effects of mercury on the breeding of the Black-crowned Night Herons at A Chau (0.3-1.2 microg/g) and Mai Po Village (0.0-1.4 microg/g). The evidence for the effects of lead and cadmium was limited but suggested there may possibly be adverse effects with lead but not cadmium. PMID- 11898802 TI - Defensive behavior and hippocampal cell proliferation: differential modulation by naltrexone during stress. AB - The present study investigated the role of endogenous opioids in the expression of defensive behaviors (DBs) and the suppression of cell proliferation (CP) in the dentate gyrus (DG) induced by exposure to predator odor, trimethyl thiazoline (TMT). Adult male rats were injected with either naltrexone (an opioid antagonist, 5 mg/kg) or saline 30 min before exposure to either TMT or a control odor. Behavior was scored for the first 15 min of odor exposure. Bromodeoxyuridine (BrdU, 200 mg/kg) was then injected, and the rats were perfused 1 hr later. Exposure to TMT increased the expression of DBs and suppressed the number of proliferating cells in the DG. Pretreatment with naltrexone attenuated the effects of TMT on DB expression but did not attenuate the effects of TMT on CP. In addition, naltrexone administration suppressed CP in the absence of TMT. These results demonstrate a dissociation between DBs and regulation of CP in the DG. PMID- 11898801 TI - Role of the dorsomedial striatum in behavioral flexibility for response and visual cue discrimination learning. AB - These experiments examined the effects of dorsomedial striatal inactivation on the acquisition of a response and visual cue discrimination task, as well as a shift from a response to a visual cue discrimination, and vice versa. In Experiment 1, rats were tested on the response discrimination task followed by the visual cue discrimination task. In Experiment 2, the testing order was reversed. Infusions of 2% tetracaine did not impair acquisition of the response or visual cue discrimination but impaired performance when shifting from a response to a visual cue discrimination, and vice versa. Analysis of the errors revealed that the deficit was not due to perseveration of the previously learned strategy, but to an inability to maintain the new strategy. These results contrast with findings indicating that prelimbic inactivation impairs behavioral flexibility due to perseveration of a previously learned strategy. Thus, specific circuits in the prefrontal cortex and striatum may interact to enable behavioral flexibility, but each region may contribute to distinct processes that facilitate strategy switching. PMID- 11898803 TI - A randomized control trial of surgical task performance in frontal recess surgery: zero degree versus angled telescopes. AB - The use of angled telescopes in frontal recess surgery has the theoretical advantage of improved visualization in areas characterized by reduced access such as the frontal recess. However, their use also is accompanied by the disadvantage of increased visuospatial distortion. To examine the surgical error and task performance of angled telescopes when compared with the use of the 0 degree telescope in frontal recess surgery, we carried out a surgical controlled trial on a cadaveric specimen. Ten surgeons performed randomly predetermined surgical tasks on both sides of the frontal recess. The surgical tasks were divided into three components (passing, grasping, and withdrawing) for analysis. Our study revealed significant difficulty passing instruments with the highly angled 70 degrees telescope as implied by the increased passing time ratio (p = 0.000). This was associated with significant risk of passing instruments blindly (p = 0.011), resulting in significant surgical error of hits to the middle turbinate (p = 0.005). This study also showed that use of less-angled telescopes (30 and 45 degrees) in frontal recess surgery does not appear to be associated with these risks. PMID- 11898804 TI - Discrepancies between medical and pharmacy records for patients on anti-HIV drugs. AB - OBJECTIVE: To compare and evaluate drug notations in outpatient medical records and in pharmacy records in a cohort of HIV-1-infected patients treated with antiretroviral drugs. METHODS: Data on 103 patients were obtained from January 1, 1998, through December 31, 1999, by medical chart review and collection of pharmacy records. Two analyses were performed. First, antiretroviral drugs and comedication in the pharmacy records were documented and compared with their appearance in the outpatient medical records. Second, a detailed comparison was performed at 5 time points during the study period for the antiretroviral drugs. Generic name, formulation, strength, and frequency of dosing as registered in the outpatient medical records were compared with those registered in the pharmacy records. RESULTS: Total drug dispensation was 1607 (366 and 1241 antiretroviral drugs and comedication, respectively). The first screening resulted in a total discrepancy of 55.1% (n = 885), of which 97.1% (n = 859) was attributed to the comedication and 2.9% (n = 26) to the antiretroviral drugs. The discrepancy for the antiretroviral drugs at the specific time points ranged from 5.1% to 12.6% when the generic name only was used, and from 7.1% to 17% when formulation, strength, and frequency of dosing were also taken into account. CONCLUSIONS: The observed discrepancy between outpatient medical records and pharmacy records mainly concerns the comedication. For the antiretroviral drugs fewer, but still substantial, discrepancies were observed. These results indicate that full exchange of information conceming drug use in this population between general practitioners and specialists (infectious disease) is lacking. PMID- 11898805 TI - Increased lithium dose requirement in a hyperglycemic patient. AB - OBJECTIVE: To report a case of apparent increased lithium clearance in a patient with persistent hyperglycemia. METHODS: Lithium doses, blood glucose concentrations, and lithium plasma concentrations were evaluated in a patient during a 53-day inpatient admission for exacerbation of bipolar disorder. The lithium dose required to increase the lithium plasma concentration by 0.1 mEq/L was computed from the lithium dose and corresponding lithium plasma concentration. This value was correlated with the blood glucose concentration. RESULTS: A plot of the lithium dose required to increase the lithium plasma concentration by 0.1 mEq/L versus the blood glucose concentration exhibited a direct linear relationship (r2 = 0.62). This plot indicates that higher lithium doses are needed during hyperglycemic states compared with euglycemic states to achieve equivalent plasma concentrations. CONCLUSIONS: Glycosuria associated with hyperglycemia induces an osmotic diuresis that increases the renal clearance of lithium, necessitating higher lithium doses to maintain therapeutic lithium plasma concentrations. PMID- 11898806 TI - Image-guided radiofrequency ablation of spinal tumors: preliminary experience with an expandable array electrode. AB - PURPOSE: Metastases to the spine are a challenging problem. Percutaneous, image guided tumor ablation with a thermal energy source, such as radiofrequency, has received increasing attention as a promising technique for the treatment of focal malignant disease. We used radiofrequency ablation for patients with unresectable, osteolytic spine metastases under computed tomographic and fluoroscopic guidance. The purpose of this study was to determine the feasibility, effectiveness, and safety of radiofrequency ablation as a palliative procedure to reduce pain and back pain-related disability in patients with vertebral and paravertebral spine tumors who were not able to benefit from radiotherapy, chemotherapy, or surgery. PATIENTS AND METHODS: Between November 1999 and January 2001, 10 patients with unresectable spine metastases were treated with radiofrequency ablation. For the ablation we used a 50-W radiofrequency generator that is connected to an expandable electrode catheter (RITA Medical System Inc., Mountain View, CA). The mean patient age was 64.4 years. Metastases were ablated in the thoracic spine, the lumbar spine, and/or the sacral bone. Tumor diameter ranged from 1.5 to 9 cm. Combined computed tomographic and fluoroscopic guidance was used to guide the procedure. Operations were carried out without heavy sedation with the patient under local anesthesia only. The thermal lesion was produced by applying temperatures of 50 degrees to 120 degrees C for 8-12 minutes. Vertebroplasty was performed in four patients by use of 3 to 5.5 mL of polymethyl methacrylate. Therapy outcome was documented by magnet resonance imaging. Before the therapy and on follow-up of an average of 5.8 months, pain was assessed with the help of the Visual Analogue Scale. Back pain-related disability was measured with the Hannover Functional Ability Questionnaire. Neurologic and health status were documented on the Frankel score and the Karnofsky index. RESULTS: At follow-up, 9 of 10 patients reported reduced pain (Visual Analogue Scale). In patients who experienced pain relief, there was an average relative pain reduction of 74.4%. Back pain-related disability was reduced by an average of 27%. Neurologic function was preserved in nine patients and improved in one. General health was stabilized in six patients, slightly increased (by 10%-20%) in two patients, significantly enhanced (by 50%) in one patient, and slightly reduced in one patient. No complications were reported. In the treated region, magnetic resonance imaging showed no further tumor growth after the therapy. DISCUSSION: Radiofrequency ablation was successfully performed in all 10 patients. Needles were placed accurately under image guidance, and a controlled lesion was created. Pain- and back pain-related disability was clearly reduced, and neurologic function was preserved or stabilized. When confirmed by further investigation, this therapy may be a new option for patients with unresectable spine tumors that do not respond to radiotherapy and chemotherapy. PMID- 11898807 TI - Serum total gangliosides and TA90-IC levels: novel immunologic markers in colorectal cancer. AB - BACKGROUND: Because of the challenge in defining prognostic markers predictive of recurrence or progression, carcinoembryonic antigen (CEA) remains the most frequently used marker in colorectal cancer, despite its low sensitivity. We hypothesized that TA90-IC status and serum ganglioside levels might be useful markers and might be of prognostic significance in colorectal cancer. METHODS: Serum samples from 68 patients undergoing surgical treatment for histologically proven colorectal cancer were analyzed for the presence of CEA, serum gangliosides, and TA90-IC. Forty-one patients had node-negative disease, whereas 27 patients had limited metastatic disease. The intent was curative resection, even for patients with metastatic disease. Cryopreserved serum specimens were analyzed in a blinded fashion for total serum ganglioside levels (by an assay that detects lipid-associated sialic acids), for CEA, and for TA90-IC (by a murine monoclonal antibody-based enzyme-linked immunosorbent assay). A positive value for TA90-IC levels was defined as an optical density (OD) of more than 0.410 at 405 nm. RESULTS: Serum ganglioside levels were elevated more frequently than CEA concentrations (84% vs 44%). The combination of serum ganglioside and CEA values was more sensitive (88%) than CEA value alone (44%) in identifying patients with early-stage colorectal cancer. TA90-IC levels were elevated more frequently than CEA concentrations (56% vs 32%). The combination of TA90-IC and CEA values was more sensitive (72%) than CEA value alone (32%) in identifying patients with advanced-stage colorectal cancer. At an enzyme-linked immunosorbent assay cutoff level of 0.410, 15 (56%) patients had positive TA90-IC values. Fourteen patients alive with residual disease had a median OD TA90-IC level of 0.879, and only three patients had levels below the OD cutoff value of 0.410. Thirteen patients with no evidence of disease had a median level of 0.277, and only four patients had OD levels > or = 0.410. TA90-IC was significantly higher in the alive with residual disease patients than those rendered no evidence of disease (P = 0.02). CONCLUSIONS: We speculate that a multiple-marker analysis that combines CEA values with serum ganglioside and TA90-IC values may be more sensitive than CEA value alone for detecting colorectal cancer. The potential prognostic significance of TA90-IC status in advanced disease warrants further investigation. PMID- 11898808 TI - Surgery for emphysema. PMID- 11898809 TI - Surgery for emphysema. PMID- 11898810 TI - Surgery for emphysema. PMID- 11898811 TI - Radiotherapy for breast cancer. PMID- 11898812 TI - Radiotherapy for breast cancer. PMID- 11898813 TI - Case 32-2001: interstitial pneumonitis and rheumatoid arthritis. PMID- 11898814 TI - A personal encounter with psychology (1937-2002). AB - An undergraduate assistantship with Abraham Maslow, research with Solomon Asch, and an indirect exposure to Ernest Nagel's philosophy of science encouraged Howard H. Kendler to become involved with methodological issues in psychology. Graduate training with Kenneth Spence led to an active research career that was initially immersed in the latent learning controversy and later, with the collaboration of his wife Tracy Kendler, in the extension of the Hull-Spence model of cognitive development. Methodological concerns from a variety of sources encouraged Kendler to express his ideas on the methodology and history of psychology as well as its role in ethical and social policy issues. A productive symbiotic relationship is created from the interaction of democracy, natural science psychology, and moral pluralism. PMID- 11898815 TI - Misconceptions about Freud's seduction theory: comment on Gleaves and Hernandez (1999). AB - D. H. Gleaves and E. Hernandez (1999) write in relation to the seduction theory that "recent writers now argue that ... Freud never made discoveries of sexual abuse" (p. 332) and that "the assertion that Freud did not make discoveries of abuse is unwarranted" (p. 324). In this article an outline of the case that Freud had no adequate grounds for his 1896 claims of having uncovered infantile "sexual scenes" is given. Some of the more important misconceptions and erroneous arguments in Gleaves and Hernandez's article are then examined. PMID- 11898816 TI - Wethinks the author doth protest too much: a reply to Esterson (2002). AB - A Esterson (2002) responded to the authors' analysis of recent reformulations of Freud's seduction theory and alleged sexual abuse discoveries. Esterson gave several additional examples of the same type of problematic writing the authors discussed in their original article. His commentary is largely a repetition of several already-published arguments, and his numerous criticisms of the article are, in the authors' opinion, without merit. The authors address confusion over inferring abuse from symptoms, treatment of symptoms versus resolution of cases, and fathers as perpetrators of abuse. It is clear that, as long as the topic of child sexual abuse elicits heated debate, so will Freud's seduction theory, but there may be times when one needs to step back to allow a debate to move forward. PMID- 11898818 TI - Certified registered nurse anesthetist. PMID- 11898817 TI - Marion White McPherson (1919-2000). PMID- 11898819 TI - Balloon kyphoplasty. PMID- 11898820 TI - Health science and national security. PMID- 11898821 TI - A South African perspective on 2001. PMID- 11898822 TI - Facts of political life. PMID- 11898823 TI - Subtle shake-up in bone-loss research. AB - Recent research in the prevention of bone loss during weightlessness is described. Scientists are studying the effects of vibration on bone loss in laboratory animals and humans. Research is focused on determining how bone formation is triggered, the effects of stimulation of the stress response in bones, and the mechanisms behind the effects. Clinton Rubin at the State University of New York at Stony Brook is experimenting with 90 Hz vibration frequencies in rats. Other researchers are studying the effects of vibration plates in the treatment of osteoporosis in postmenopausal women and in children with low bone density. Tests among astronauts have been proposed, but none are scheduled. PMID- 11898824 TI - Letting thoughts take wing. AB - Recent developments in neuroelectronics are applied to aviation and airplane flight control instruments. Electromyographic control has been applied to flight simulations using the autopilot interface in order to use gestures to give bank and pitch commands to the autopilot. In other demonstrations, direct rate control was used to perform repeated successful landings and the damage-adaptive capability of inner-loop neural and propulsion-based controls was utilized. PMID- 11898826 TI - Maverick medicine. AB - Since Western medics discovered the benefits of acupuncture and other Chinese healing methods, lawmakers have had a tough time fitting it all into the traditional model. PMID- 11898827 TI - Heart disease and stroke still leading killers. PMID- 11898829 TI - Improved organ donor laws desperately needed. PMID- 11898830 TI - Life sciences and space research X. Proceedings of the Open Meeting of Working Group 5 of the Fourteenth Plenary Meeting of Cospar, Seattle, Washington, USA, 21 June-2 July, 1971. PMID- 11898831 TI - Influence of Cosmos 368 space flight conditions on radiation effects in yeasts, hydrogen bacteria and seeds of lettuce and pea. AB - Radiobiological studies have been carried out on board the Cosmos 368 satellite, launched on 8 October 1970 and returning to earth on 14 October 1970. Yeast diploid cells Saccharomyces ellipsoides, Megri strain 139-B, haploid cells Zygosaccharomyces Baili, hydrogen bacteria Hydrogenomonas eutropha, strain Z-1, Berlin variety lettuce seeds and Capital variety pea seeds were used in these experiments. The biological specimens were irradiated with gamma-rays at dose rates of 71.8 and 6.7 rad d-1 suring the packaging of containers and after returning the samples to the laboratory. It was found that both on pre-radiation and post-radiation exposure space flight factors did not greatly influence radiobiological effects. PMID- 11898832 TI - OFO experimental techniques and preliminary conclusions: is artificial gravity needed during prolonged weightlessness? AB - The technique of single unit recording from body systems generating electrical pulses coherent with their basic function (CNS, muscles, sense organs) has been proved feasible during the OFO A orbital flight, an automatic physiological experiment. All microelectrode implants survived the lift off of a Scout vehicle. The far-reaching impact of such a technique in biological space research and in the laboratory is discussed. The results of recording 155 hours of orbital flight of pulses from the nerve fibres of four vestibular gravity sensors in two bull frogs indicate that the vestibular organ adjusts to zero g. As all the other biological changes observed during orbit are due to lack of exercise, it is concluded that artificial gravity might not be necessary during prolonged space missions or on low gravity celestial bodies. PMID- 11898834 TI - Physiological and hematological effects of chronic irradiation. AB - The information available for assessing radiation hazard involved in prolonged space missions is so far insufficient. Therefore, a three-year experiment was carried out in which 180 dogs were exposed to irradiation, simulating the dose value and rate of exposure that may occur in a real space flight of long duration. The exposure included a chronic irradiation (with dose rates of 21, 62 and 125 rads/year) and a combined irradiation during which the animals were exposed to chronic and acute irradiations with a dose of 8 or 42 rads applied three times every year, the annual total dose being 120 or 188 rads, respectively. Insignificant hematopoietic changes, e.g. the ratio of red to white blood cells, their decrease in the peripheral blood, were found. Distinct changes in the reproductive function were noted. The general condition of the animals was satisfactory. Most animals endured well an additional physical load and retained conditioned reflexes developed before irradiation. The experimental findings suggest that the effects are early symptoms of the first stage of chronic radiation damage. PMID- 11898833 TI - Test of a life support system with Hirudo medicinalis in a sounding rocket. AB - Two Nike-Tomahawk rockets each carrying two Biosondes were launched from Wallops Island, Virginia, the first on 10 December 1970 and the second on 16 December 1970. The primary objective of both flights was to test the Biosonde life support system under a near weightless environment and secondarily to subject the Hirudo medicinalis to the combined stresses of a rocket flight. The duration of the weightless environment was approximately 6.5 minutes. Data obtained during the flight by telemetry was used to ascertain the operation of the system and the movements of the leeches during flight. Based on the information obtained, it has been concluded that the operation of the Biosondes during the flight was similar to that observed in the laboratory. The experiment and equipment are described briefly and the flight results presented. PMID- 11898835 TI - The effects of dose protraction on hematopoiesis in the primate and dog. AB - The modifying effects of dose protraction by fractionation or continuous low dose rate exposure are not well known in the primate. Comparative studies between the two mammalian species dog and monkey with widely differing acute LD50 values have not been made. A program to study dose-rate effects on injury and recovery of the bone marrow in dogs and monkeys under simultaneous and similar exposure conditions is in progress. The equivalent residual dose (ERD) assumptions of 10% irreparable injury and a 28-day recovery half-time for the reparable portion (90%) of radiation were studied in dogs (beagles) and monkeys (Macaca arctoides) using variable recovery times and appropriate fractionated exposures to attain but not exceed theoretical effective residual body burdens of 200 rads over a 1 year period. The ERD assumptions were tolerated by the dogs but resulted in death to 7 of 8 monkeys tested. The same ERD assumptions were tested with monkeys (Macaca mulatta) using a fixed recovery time span and an upper ERD limit of 100 rads. Monkeys in this study have tolerated 1900 rads of gamma-ray exposure (approximately 3 times the acute lethal dose) with minimal suppression of the hematopoietic system. PMID- 11898836 TI - Summary of latent effects in long term survivors of whole body irradiations in primates. AB - The USAF School of Aerospace Medicine, Radiobiology Division, Brooks Air Force Base, Texas presently is maintaining a colony of over 450 primates in which the whole body has been exposed to various types of space radiation including protons and electrons. The majority of the primates (Macaca mulatta) were exposed during 1965. Types of radiation involved are 2 MeV X-rays, 5 MeV-2.3 GeV protons and 1.6 MeV electrons. Low energy proton dose range up to 3000 rad (50-100 rad min-1) whereas the penetrating energy doses range up to 700 rad (15-100 rad min-1). Primates from a simulated solar flare exposure are also included. In late 1970, a small group of primates exposed to 108 and 85 MeV alpha particles (eye and partial body only) were added to the colony. Data are available in the following areas: (i) chronic skin changes; (ii) testicular atrophy; (iii) cataractogenesis; (iv) hematological and serum biochemical analysis; (v) incidence of tumors; (vi) causes of death; (vii) body weight variations; and (viii) summary of alpha particle experiences. PMID- 11898837 TI - Analysis of survival and cause of death statistics for mice under single and duration-of-life gamma irradiation. AB - The late effects of protracted exposure to low levels of external radiation continue to be a matter of operational concern in long-range space flight. Studies have been carried out on young adult mice exposed to daily levels of 60Co gamma irradiation ranging from 0.3 to over 30 R day-1. The lowest level is comparable with the occupational maximum permissible dose for the atomic energy industry. There is little evidence of life shortening at that level, but as exposure increases, there is an exponential decline in life expectancy. The life shortening coefficient is approximately 4 days/100 R accumulated or 4% R-1 day-1. When life-shortening is < or = 15%, all of the increased mortality can be attributed to radiation-induced increases in death rates from neoplastic diseases, including various forms of leukemia and pulmonary tumors. Age-specific death rates for mice dying of all other causes remain the same as the controls throughout life, at the lowest doses. A non-neoplastic disease component of excess mortality rate emerges at 6 R day-1 and above. The risk of death from all and specific causes following single exposures compared with protracted lifetime irradiation shows a clear effect of protraction. Leukemia death rates are reduced by a factor of 5 or more at all daily exposure levels below 20-30 R day-1. Risks for other causes of death are also reduced, but to a variable degree. PMID- 11898838 TI - Effects of simulated space vacuum on bacterial cells. AB - The effect of vacuum on bacterial cells is related to water desorption. Below water vapour pressure the inactivation remains constant, independent of total pressure and exposure time. In subsequent growth, the lag-phase of the survivors is delayed. Combined treatment with vacuum and radiation (X-rays or uv of 254 nm wavelength) results in synergistic effects, whereas vacuum and heat can act antagonistically. The vacuum inactivated cells (indicated as loss of colony forming ability) are completely damaged. They do not show cellular elongation, phage production or respiration. The cellular membrane becomes permeable by vacuum exposure: biomolecules are released from the cells when re-suspended after vacuum treatment. PMID- 11898839 TI - Biological instrumentation for the Viking 1975 mission to Mars. AB - A brief introduction is given on why Mars is of interest from a biological point of view, along with an overview of the Viking 1975 mission. Details are given about the four biology instruments aboard the spacecraft and the experiments for which they are to be used. These are: the carbon assimilation experiment to determine whether the soil is biologically active, by incubation in presence of 14C-labelled CO and CO2 (known to be present in the Martian atmosphere); the label release experiment to detect metabolic activity by the release of radioactive CO2, from 14C-labelled simple organic substrates; the gas exchange experiment to detect biological activity by repeated gas chromatography analysis of soil samples; the light scattering experiment, where increase of scattering and decrease of light transmission would indicate the growth of organisms. Examples are given of data obtained with terrestrial soils in these experiments. PMID- 11898840 TI - An integrated multi-purpose biology instrument utilizing a single detector, the mass spectrometer. AB - A mass spectrometer is used to analyze the gas phase in a number of reaction vessels filled with Martian soil. By choosing appropriate incubation conditions this instrument can be used to perform a wide spectrum of experiments ranging from the observation of general indices of life, i.e. processes and patterns unexplainable by physico-chemical mechanisms, to assays utilizing isotopes which probe for specific metabolic processes. Of particular interest is the in situ incubation in which a Martian soil sample is maintained at a constant temperature and its gas phase composition analyzed with time. Properly interpreted, this is a very general life-detection probe which makes minimal assumption as to the nature of Martian biology. Other assays and measurements concerning the soil and the atmosphere compatible with this method are also described. PMID- 11898841 TI - Effects of weightlessness on astronauts--a summary. AB - This paper reviews the adaptive changes observed in the United States astronauts during flight programs to this date. A series of postulates are offered as to what is happening in these adaptive events. A hypothesis is proposed as to the interrelationship of events observed in the body systems and functions involved. The importance of undertaking an extensive life sciences program. including an on orbit phase of study as well as pre- and post-flight studies is discussed. Finally, the role the Skylab flight plays in the United States Space Program in achieving the future requirements for more extensive life sciences data is summarized. PMID- 11898842 TI - Effects of an 18-day flight on the human body. AB - During their flight on board the Soyuz 9 A.G. Nikolayev and V.I. Sevastyanov adapted to weightlessness by the 3rd-4th day and stabilized their physiological functions by the end of the mission. During flight both cosmonauts maintained normal performance, sleep and appetite; they readily developed a new stereotype of movements and exhibited no noticeable increase of the circulatory function in response to a standard physical load. In contrast to the effects of shorter term flights, this mission "used unusual and distressing feelings in the crew members aggravated by distinct changes in the major- physiological systems during the first day of recovery. In the immediate postflight hours the transition from the recumbent to the sitting position brought about circulation disorders; 24 hours later the cosmonauts still walked with uncertainty and kept the erect position at rest on account of a significant elevation of their centre of gravity. Weight losses, shifts of water and mineral metabolism, bone tissue demineralization and symptoms of orthostatic intolerance observed in this flight were similar to those resulting from earlier short-term missions. Of importance was a dysbacteriotic change in the skin and nasal microflora. Physiological changes in the Soyuz 9 crew members were functional and reversible, being on the whole in agreement with predicted effects. These results call for the development of specific measures facilitating the post-flight adaptation of space pilots in view of future long duration space flights. PMID- 11898843 TI - Functional insufficiency of the neuromuscular system caused by weightlessness and hypokinesia. AB - The results of study of the crew members of the spaceships Soyuz are described, and the effects of weightlessness on reflex excitability, muscular tone and muscle contractibility discussed. A certain decrease in postural muscular tone and strength, increase in reflex excitability at rest and increase in bioelectric activity of muscles at work has been found in the cosmonauts after their stay in a weightless environment. The circumference of the lower extremities decreased. PMID- 11898844 TI - Studies on weightlessness in a primate in the Biosatellite 3 experiment. AB - In June 1969 a male Macaca nemestrina (pigtail macaque) was flown in earth orbit for 8.8 days in NASA Biosatellite 3. The experiment examined in detail central nervous and cardiovascular functions, and included pre- and post-flight whole body metabolic assessment, in-flight urine analysis, and pre-and post-flight bone density measurements. Although the sleep/wake cycle was 24 hr, a phase angle lag of 2 hr from the imposed night/day mode occurred. A definite desynchronosis occurred, with rhythms longer than 24 hr in pCO2, brain and body temperature and heart rate, although arterial blood pressure remained at 24 hr. Sleep states were remarkably fragmented and unusually brief in duration. Vestibular and ocular disturbances were evident. These changes began concurrently with onset of weightlessness and were not secondary to altered fluid balance or body temperature. Sleep patterns lie between those of normal man and man with high cervical cord transection. There was an immediate and sustained increase in central venous pressure in weightlessness and this is considered to have initiated a Henry-Gauer reflex which initially maintained a high urine volume. This, coupled with a high evaporative fluid loss, produced an early dehydration probably associated with electrolyte imbalances. Body weight was 20% lower at recovery than at launch. Ventricular fibrillation supervened 8 hr after recovery. PMID- 11898845 TI - Molecular basis of gestational trophoblastic diseases. AB - Gestational trophoblastic disease (GTD) encompasses a diverse group of lesions with specific pathogenesis, morphological characteristics and clinical features. The modified World Health Organization-classification of GTD includes complete and partial hydatidiform mole, invasive mole, choriocarcinoma, placental site trophoblastic tumor, epithelioid trophoblastic tumor, exaggerated placental site, and placental site nodule. The various forms of gestational trophoblastic disease can be defined and related to discrete pathologic aberrations occurring at different stages of trophoblastic differentiation. Some of these lesions are true neoplasms, whereas others represent abnormally formed placentas with a predisposition for neoplastic transformation of the trophoblast. Except hydatidiform moles in which the cytogenetic studies have been extensively reported, the pathogenesis of other trophoblastic lesions is poorly understood. Recent studies have shed light on the molecular mechanisms of trophoblastic function, especially as it relates to trophoblastic disease. This review will focus on these advances with special emphasis on the pathogenesis of each specific form of GTD. In addition, the morphology and clinical behavior of each of these entities will be briefly discussed. PMID- 11898846 TI - Homeobox genes and human genetic disorders. AB - Homeobox genes encode transcriptional regulators of embryonic development. Many genetic disorders affecting multiple organ systems have been associated with a diverse array of mutations in one or another of at least 27 different members of this gene family. We briefly describe the affected genes and the major phenotypes presented by the patients that carry the mutations. Although cause-and-effect relationships are difficult to prove in human genetics, there is little doubt that the observed mutations play a crucial role in the etiology of the associated disorders. The impressive wealth of collected data greatly benefits genetic counseling and stimulates efforts to develop novel avenues of targeted therapy. PMID- 11898847 TI - Molecular aspects of sex differentiation. AB - Mammalian sex differentiation involves the action of a cascade of genes. Discovery of the sex-determining region of the Y chromosome (SRY) marked the beginning of the delineation of the genes in the cascade. Studies of the genetics of mammalian sex reversal and the embryogenesis of the mice are essential in this endeavor. A number of genes involved in the pathway have been identified and all except one of these genes have a putative role in male sex differentiation. Besides SRY being the master switch in male sex differentiation the hierarchical relationship of the genes identified are far from being understood. Similarly, our knowledge of the genetic regulation of female sex differentiation is minimal. Differential screening and gene expression profiling bring a new dimension to the pursuit with the identification of a number of genes previously unknown to be involved in sex differentiation. Wider application of functional genomic techniques and introduction of proteomic analyses are expected to shed light to our understanding of this complicated developmental process. PMID- 11898848 TI - Formation and malformation of the vertebrate left-right axis. AB - Despite an externally symmetric body plan, the internal viscera of all vertebrates are asymmetric with respect to the left-right body axis. Determination of the handedness of this asymmetry is nonrandom and highly conserved among vertebrates. Errors in patterning along the left-right axis, which occur in about 1 in 10,000 human births, may result in significant morbidity and mortality. During early embryonic development, midline structures, in particular the node, coordinate patterning of the three main embryonic axes: anterior-posterior, dorsal-ventral, and left-right. A current model for specification of the handedness of left-right axis asymmetry invokes the activity of embryonic cilia in the node that create a net leftward flow of extraembryonic fluid. This flow is proposed to provide a signal for subsequent asymmetric gene expression. Signaling from the node defines patterns of asymmetric gene expression on the left and right sides of the embryo. These signals for "left" and "right" are ultimately interpreted by organ primordia during later development. Complex activating and inhibiting interactions involving TGF-beta family members, as well as homeobox transcription factors, mediate these asymmetric patterns of gene expression. The identification of the genes regulating left-right axis patterning in model organisms has resulted in the characterization of human mutations associated with left-right axis malformations. PMID- 11898849 TI - Hedgehog signaling in gastrointestinal development and disease. AB - The development of the gastrointestinal (GI) tract and its associated parenchymal organs depends on Hedgehog signals from the endoderm to the surrounding mesoderm. During development, Hedgehog signaling is essential for patterning the GI tract along anterior-posterior (A-P), dorsal-ventral (D-V), and radial axes, as well as in maintenance of stem cells. Our knowledge about these roles for Hedgehog signaling is derived from studies of developmental defects that result from disrupted or activated Hedgehog signaling in model organisms including mouse, chick, and frog. These studies provide evidence for distinct roles of specific Hedgehog ligands in GI development. Studies in model organisms have also elucidated how Hedgehog signaling may function in development and function of the GI tract in humans. Several diseases and congenital syndromes are known to result from genetic defects in Hedgehog signaling components, and this pathway may ultimately prove to be an important target for future diagnostic and therapeutic tools. PMID- 11898850 TI - Novel treatment for neuronopathic lysosomal storage diseases--cell therapy/gene therapy. AB - Most lysosomal storage diseases (LSD) exhibit neurological symptoms and there has been limited success in their treatment. Innovative treatments employing novel therapy or gene therapy may offer the prospect of improvement. Recent attempts to treat the neurological forms of LSD include neural stem cell therapy, mesenchymal stem cell therapy, hematopoietic stem cell therapy and gene therapy. Additional approaches have included substrate deprivation/chaperone therapy for the treatment of LSD. This article reviews these new technologies, discusses recent progress, and suggests their possible application. PMID- 11898852 TI - Molecular analysis of complex tissues is facilitated by laser capture microdissection: critical role of upstream tissue processing. AB - Every tissue contains heterogeneous cell populations. Laser capture microdissection (LCM) facilitates cell isolation from complex tissues followed by molecular analysis. LCM entails placing a transparent film over a tissue section or a cytological sample, visualizing the cells microscopically, and selectively adhering the cells of interest to the film with a focused pulse from an infrared laser. The film with the procured cells is then removed from the original sample and placed directly into DNA, RNA, or protein-extraction buffer for processing. LCM has revolutionized molecular analysis of complex tissues because it combines the topographic precision of microscopy with the power of molecular genetics, genomics, and proteomics. However, the success of molecular analysis still depends on the experimental design and requires the understanding of each technical step involved in specimen preparation. This review attempts to rationalize and demystify the choice of various technical options in upstream tissue processing supporting global analytical strategies. PMID- 11898851 TI - Mechanotransduction in Caenorhabditis elegans: the role of DEG/ENaC ion channels. AB - One of the looming mysteries in signal transduction today is the question of how mechanical signals, such as pressure or mechanical force delivered to a cell, are interpreted to direct biological responses. All living organisms, and probably all cells, have the ability to sense and respond to mechanical stimuli. At the single-cell level, mechanical signaling underlies cell-volume control and specialized responses such as the prevention of poly-spermy in fertilization. At the level of the whole organism, mechanotransduction underlies processes as diverse as stretch-activated reflexes in vascular epithelium and smooth muscle; gravitaxis and turgor control in plants; tissue development and morphogenesis; and the senses of touch, hearing, and balance. Intense genetic, molecular, and elecrophysiological studies in organisms ranging from nematodes to mammals have highlighted members of the recently discovered DEG/ENaC family of ion channels as strong candidates for the elusive metazoan mechanotransducer. Here, we discuss the evidence that links DEG/ENaC ion channels to mechanotransduction and review the function of Caenorhabditis elegans members of this family called degenerins and their role in mediating mechanosensitive behaviors in the worm. PMID- 11898853 TI - Regulation of L-arginine transport and metabolism in vascular smooth muscle cells. AB - L-Arginine is a semi-essential amino acid that is metabolized to important regulatory molecules. L-Arginine is transported into vascular smooth muscle cells (SMC) by the cationic amino acid transporter (CAT) family of proteins where it is metabolized to nitric oxide (NO), polyamines, or L-proline. Inflammatory mediators, growth factors, and hemodynamic forces stimulate the transport of L arginine in vascular SMC by inducing CAT gene expression. However, they exert highly specific and divergent regulatory effects on L-arginine metabolism. Inflammatory cytokines induce the expression of inducible NO synthase (iNOS) and direct the metabolism of L-arginine to the antiproliferative gas, NO. In contrast, growth factors stimulate the expression of arginase I and ornithine decarboxylase (ODC) and channel the metabolism of L-arginine to growth stimulatory polyamines. Alternatively, cyclic mechanical strain blocks both iNOS and ODC activity and stimulates arginase I gene expression, directing the metabolism of L-arginine to the formation of L-proline and collagen. Thus, specific biochemical and biophysical stimuli that are found in the circulation regulate the transport and metabolism of L-arginine in vascular SMC. The ability of these physiologically relevant stimuli to upregulate L-arginine transport and generate specific L-arginine metabolites modulates SMC function and may influence the development of vascular disease. PMID- 11898854 TI - Predicted structures of two proteins involved in human diseases. AB - Structures of 79 proteins involved in human diseases were predicted by sequence alignments with structural templates. The predicted structures for ALDP and CSA, proteins responsible for adrenoleukodystrophy and the Cockayne syndrome, respectively, were analyzed to elucidate the molecular basis of disease mutations. In particular we positioned residue P484 of ALDP in the homodimer interface. This positioning is consistent with a recent experimental finding that the mutation P484R significantly decreases the self-interaction of ALDP and suggests that the disease mechanism of this mutation lies in the impaired ALDP dimerization. We identified two new WD repeats in CSA and suggest that one of these forms part of the interaction surface with other proteins. PMID- 11898855 TI - The local Ca concentration profile in the vicinity of a Ca channel. AB - Intracellular Ca signaling is governed by diffusion and buffering of Ca ions. Mobile endogenous buffers increase the redistribution of Ca ions and tend to restrict the regions of elevated Ca concentrations to the close vicinity of the channel. These Ca microdomains dominate fast Ca signaling, as observed, e.g., in synaptic transmission. The steady-state solution of the linearized differential equations of buffered Ca diffusion, as developed by Neher and Naraghi and Neher, will be reviewed and generalized to the case of more than two buffers. Immobile buffers do not enter the steady-state equations, but instead slow down Ca diffusion and prolong the time to reach the steady state. Based on this phenomenon, a quite different putative mechanism to localize Ca will be suggested that is likely to be realized in photoreceptors where Ca source, Ca sink, and Ca sensor form a complex, as was recently reported. PMID- 11898856 TI - Catalytic antibodies: structure and function. AB - More than 10 years have now elapsed since the first reports confirmed that antibodies could be programmed as catalysts for chemical processes. Much of the initial research focussed on exploring the scope and utility of these new biocatalysts. Recently however, increasing information gleaned from X-ray analyses is allowing an exciting insight into the structural basis of antibody catalyzed reactions. This review details the evolving knowledge of the structure function relationship for catalytic antibodies that accelerate a range of different reaction classes. PMID- 11898857 TI - How can the differences among AT1-receptor antagonists be explained? AB - Over the last few years we have seen a new class of antihypertensive drug evolve, the angiotensin II subtype 1 receptor antagonists. Hypothetically, all substances in this class should have the same effect on blood pressure and on end-organ damage as they all block the AT1 receptor. However, there are distinctions between them that may explain the significant and clinically important differences that seem to exist within this class of drug. An explanation for the differences may be found in receptor-antagonist kinetics. The receptor-antagonist interaction may be fitted to a two-state, two-step model which determines how large a part of the binding that will be surmountable and how large a part that will be insurmountable. The proportion of surmountable/insurmountable binding fits nicely to the duration of binding of the antagonist to the receptor, which may be translated into efficacy for the antagonist as outlined in the following review. PMID- 11898858 TI - Determining the three-dimensional fold of a protein from approximate constraints: a simulation study. AB - We propose a new approach for calculating the three-dimensional (3D) structure of a protein from distance and dihedral angle constraints derived from experimental data. We suggest that such constraints can be obtained from experiments such as tritium planigraphy, chemical or enzymatic cleavage of the polypeptide chain, paramagnetic perturbation of nuclear magnetic resonance (NMR) spectra, measurement of hydrogen-exchange rates, mutational studies, mass spectrometry, and electron paramagnetic resonance. These can be supplemented with constraints from theoretical prediction of secondary structures and of buried/exposed residues. We report here distance geometry calculations to generate the structures of a test protein Staphylococcal nuclease (STN), and the HIV-1 rev protein (REV) of unknown structure. From the available 3D atomic coordinates of STN, we set up simulated data sets consisting of varying number and quality of constraints, and used our group's Self Correcting Distance Geometry (SECODG) program DIAMOD to generate structures. We could generate the correct tertiary fold from qualitative (approximate) as well as precise distance constraints. The root mean square deviations of backbone atoms from the native structure were in the range of 2.0 A to 8.3 A, depending on the number of constraints used. We could also generate the correct fold starting from a subset of atoms that are on the surface and those that are buried. When we used data sets containing a small fraction of incorrect distance constraints, the SECODG technique was able to detect and correct them. In the case of REV, we used a combination of constraints obtained from mutagenic data and structure predictions. DIAMOD generated helix loop-helix models, which, after four self-correcting cycles, populated one family exclusively. The features of the energy-minimized model are consistent with the available data on REV-RNA interaction. Our method could thus be an attractive alternative for calculating protein 3D structures, especially in cases where the traditional methods of X-ray crystallography and multidimensional NMR spectroscopy have been unsuccessful. PMID- 11898859 TI - Characterization of reconstituted Fo from wild-type Escherichia coli and identification of two other fluxes co-purifying with Fo. AB - We purified the ATPase Fo sector from a nonoverexpressing strain of Escherichia coli, reconstituted it into lipid vesicles made of either asolectin or two different mixtures of purified lipids, and measured proton flux through the reconstituted proton channel. We measured single-channel conductances and found that Fo activity depends on both lipids and reconstitution methods. In asolectin vesicles, Fo has a single-channel conductance of about 0.2 fS. Additionally, the relatively impure Fo prepared from cells carrying single-copy ATPase genes allowed us to observe two other fluxes, a nonselective cation leak (C(L)) and a slow H+ flux (Hs). Unlike the Fo flux, these fluxes could not be blocked by the Fo inhibitor DCCD. The C, reduces the total apparent trapped volume inside vesicles and therefore must equilibrate both H+ and K+ in the vesicles that contain it. When reconstituted into bilayers, these Fo preparations displayed a 120 pS cation channel with characteristics consistent with C(L) flux. The Hs conducts only H+ but at a slower rate than the Fo. We were therefore able to: 1) quantitate the single-channel conductance of the Fo, 2) demonstrate that our Fo purification method co-purified other membrane proteins that have ion-conduction properties, and 3) show that certain lipids are necessary for functional reconstitution of Fo. PMID- 11898860 TI - Mechanosensitive channel of Thermoplasma, the cell wall-less archaea: cloning and molecular characterization. AB - By using a functional approach of reconstituting detergent-solubilized membrane proteins into liposomes and following their function in patch-clamp experiments, we identified a novel mechanosensitive (MS) channel in the thermophilic cell wall less archaeon Thermoplasma volcanium. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the enriched protein fractions revealed a band of approx 15 kDa comparable to MscL, the bacterial MS channel of large conductance. 20 N-terminal residues determined by protein microsequencing, matched the sequence to an unknown open reading frame in the genome of a related species Thermoplasma acidophilum. The protein encoded by the T. acidophilum gene was cloned and expressed in Escherichia coli and reconstituted into liposomes. When examined for function, the reconstituted protein exhibited properties typical of an MS ion channel: 1) activation by negative pressure applied to the patch-clamp pipet, 2) blockage by gadolinium, and 3) activation by the anionic amphipath trinitrophenol. In analogy to the nomenclature used for bacterial MS channels, the MS channel of T acidophilum was termed MscTA. Secondary structural analysis indicated that similar to MscL, the T. acidophilum MS protein may have two transmembrane domains, suggesting that MS channels of thermophilic Archaea belong to a family of structurally related MscL-like ion channels with two membrane spanning regions. When the mscTA gene was expressed in the mscL- knockout strain and the MscTA protein reconstituted into liposomes, the gating of MscTA was characterized by very brief openings of variable conductance. In contrast, when the mscTA gene was expressed in the wild-type mscL+ strain of E. coli, the gating properties of the channel resembled MscL. However, the channel had reduced conductance and differed from MscL in its kinetics and in the free energy of activation, suggesting that MscTA and MscL can form functional complexes and/or modulate each other activity. Similar to MscL, MscTA exhibited an increase in activity in liposomes made of phospholipids having shorter acyl chain, suggesting a role of hydrophobic mismatch in the function of prokaryotic MS channels. PMID- 11898861 TI - Mechanosensitive channels in archaea. AB - The ubiquity of mechanosensitive (MS) channels triggered a search for their functional homologues in Archaea, the third domain of the phylogenetic tree. Two types of MS channels have been identified in the cell membranes of Haloferax volcanii using the patch clamp technique. Recently MS channels were identified and cloned from two archaeal species occupying different environmental habitats. These studies demonstrate that archaeal MS channels share structural and functional homology with bacterial MS channels. The mechanical force transmitted via the lipid bilayer alone activates all to date known prokaryotic MS channels. This implies the existence of a common gating mechanism for bacterial as well as archaeal MS channels according to the bilayer model. Based on recent evidence that the bilayer model also applies to eukaryotic MS channels, mechanosensory transduction probably originated along with the appearance of the first life forms according to simple biophysical principles. In support of this hypothesis the phylogenetic analysis revealed that prokaryotic MS channels of large and small conductance originated from a common ancestral molecule resembling the bacterial MscL channel protein. Furthemore, bacterial and archaeal MS channels share common structural motifs with eukaryotic channels of diverse function indicating the importance of identified structures to the gating mechanism of this family of channels. The comparative approach used throughout this review should contribute towards understanding of the evolution and molecular basis of mechanosensory transduction in general. PMID- 11898862 TI - Fluorescence correlation spectroscopy and its potential for intracellular applications. AB - Fluorescence correlation spectroscopy (FCS) is a time-averaging fluctuation analysis of small molecular ensembles, combining maximum sensitivity with high statistical confidence. Among a multitude of physical parameters that are, in principle, accessible by FCS, it most conveniently allows to determine local concentrations, mobility coefficients, and characteristic rate constants of fast reversible and slow-irreversible reactions of fluorescently labeled biomolecules at very low (nanomolar) concentrations, under equilibrium conditions and without physical separation. Its presently most popular instrumentation by confocal microscope setups allows for a spatial resolution of fractions of femtoliters for the measurement volumes, containing sparse or even single molecules at any time, and encourages the adaptation of the solution-based technique for cellular applications. The scope of this review is thus, to introduce the FCS technique in particular to the reader with biological background, searching for new methods for a precise quantification of physical parameters governing cellular mechanisms and dynamics, especially if high sensitivity and fast dynamic resolution are required. After a short theoretical introduction, examples are given for the so far most important experimental applications, with respect to their implementation in cellular systems. As an interesting alternative to the confocal instrumentation, two-photon excitation will be introduced, offering a number of important advantages especially in cellular systems with high-noise and low signal levels. PMID- 11898863 TI - Programming the Drosophila embryo 2: from genotype to phenotype. AB - Although development is a single hierarchical process, scientists tend to study only one level at a time: molecular, cellular, or organismal. The data and theory are available to integrate molecular, cellular, and organismal levels into a series of maps for development of the Drosophila embryo. These maps link the transcriptional cascade with mitotic and phenotypic fate maps to trace hierarchical mechanisms of development from the genotype in the egg to the phenotype in the larva. PMID- 11898864 TI - Structure of the vacuolar adenosine triphosphatases. AB - Vacuolar adenosine triphosphatases (V-ATPases) represent an important class of proton pumps found in endomembrane systems of eucaryotic cells, where they are involved in pH regulation. Progress has been made in the structure determination of this large, membrane-bound multisubunit enzyme complex. Electron microscopy of the V-ATPase has revealed a ball-and-stalk-like structure similar to F1F0-type ATP synthase, to which the V-ATPase is evolutionary related. Aside from the overall structural similarity of the V-ATPase and F-ATP synthase, a number of distinct structural differences exist between the two related enzymes, giving clues to their different function and regulation in the organism. PMID- 11898866 TI - Biochemical reactivity of melatonin with reactive oxygen and nitrogen species: a review of the evidence. AB - Melatonin (N-acetyl-5-methoxytryptamine), an endogenously produced indole found throughout the animal kingdom, was recently reported, using a variety of techniques, to be a scavenger of a number of reactive oxygen and reactive nitrogen species both in vitro and in vivo. Initially, melatonin was discovered to directly scavenge the high toxic hydroxyl radical (*OH). The methods used to prove the interaction of melatonin with the *OH included the generation of the radical using Fenton reagents or the ultraviolet photolysis of hydrogen peroxide (H202) with the use of spin-trapping agents, followed by electron spin resonance (ESR) spectroscopy, pulse radiolysis followed by ESR, and several spectrofluorometric and chemical (salicylate trapping in vivo) methodologies. One product of the reaction of melatonin with the *OH was identified as cyclic 3 hydroxymelatonin (3-OHM) using high-performance liquid chromatography with electrochemical (HPLC-EC) detection, electron ionization mass spectrometry (EIMS), proton nuclear magnetic resonance (1H NMR) and COSY 1H NMR. Cyclic 3-OHM appears in the urine of humans and other mammals and in rat urine its concentration increases when melatonin is given exogenously or after an imposed oxidative stress (exposure to ionizing radiation). Urinary cyclic 3-OHM levels are believed to be a biomarker (footprint molecule) of in vivo *OH production and its scavenging by melatonin. Although the data are less complete, besides the *OH, melatonin in cell-free systems has been shown to directly scavenge H2O2, singlet oxygen (1O2) and nitric oxide (NO*), with little or no ability to scavenge the superoxide anion radical (O2*-) In vitro, melatonin also directly detoxifies the peroxynitrite anion (ONOO-) and/or peroxynitrous acid (ONOOH), or the activated form of this molecule, ONOOH*; the product of the latter interaction is proposed to be 6-OHM. How these in vitro findings relate to the in vivo antioxidant actions of melatonin remains to be established. The ability of melatonin to scavenge the lipid peroxyl radical (LOO*) is debated. The weight of the evidence is that melatonin is probably not a classic chain-breaking antioxidant, since its ability to scavenge the LOO* seems weak. Its ability to reduce lipid peroxidation may stem from its function as a preventive antioxidant (scavenging initiating radicals), or yet unidentified actions. In sum, in vitro melatonin acts as a direct free radical scavenger with the ability to detoxify both reactive oxygen and reactive nitrogen species; in vivo, it is an effective pharmacological agent in reducing oxidative damage under conditions in which excessive free radical generation is believed to be involved. PMID- 11898865 TI - Transient, highly populated, building blocks folding model. AB - Protein folding is a hierarchical event, in which transiently formed local structural elements assemble to yield the native conformation. In principle, multiple paths glide down the energy landscape, but, in practice, only a few of the paths are highly traveled. Here, the literature is reviewed in this light, and, particularly, a hierarchical, building block protein-folding model is presented, putting it in the context of a broad range of experimental and theoretical results published over the past few years. The model is based on two premises: First, although the local building block elements may be unstable, they nevertheless have higher population times than all alternate conformations; and, second, protein folding progresses through a combinatorial assembly of these elements. Through the binding of the most favorable building block conformers, there is a redistribution of the conformers in solution, propagating the protein folding reaction. We describe the algorithm, and illustrate its usefulness, then we focus on its utility in assigning simple vs complex folding pathways, on chaperonin-assisted folding, on its relevance to domain-swapping processes, and on its relevance and relationship to disconnectivity graphs and tree diagrams. Considering protein folding as initiating from local transient structural elements is consistent with available experimental and theoretical results. Here, we have shown that, early in the folding process, sequential interactions are likely to take place, even if the final native fold is a complex, nonsequential one. Such a route is favorable kinetically and entropically. Through the construction of anatomy trees, the model enables derivation of the major folding pathways and their bumps, and qualitatively explains the kinetics of protein folding. PMID- 11898867 TI - Fluorescent pteridine nucleoside analogs: a window on DNA interactions. AB - Pteridine nucleoside analog probes are highly fluorescent and offer different approaches to monitor subtle DNA interactions with other molecules. Similarities in structure and size to native nucleosides make it possible to incorporate these probes into oligonucleotides through the standard deoxyribose linkage. These probes are formulated as phosphoramidites and incorporated into oligonucleotides using automated DNA synthesis. Their position within the oligonucleotide renders them exquisitely sensitive to changes in structure as the oligonucleotide meets and reacts with other molecules. Changes are measured through fluorescence intensity, anisotropy, lifetimes, spectral shifts, and energy transfer. The fluorescence properties of pteridine nucleoside analogs as monomers and incorporated into single and double stranded oligonucleotides are reviewed. The two guanosine analogs, 3MI and 6MI, and two adenosine analogs, 6MAP and DMAP, are reviewed in detail along with applications utilizing them. PMID- 11898868 TI - The diagnosis of disorders caused by hand-transmitted vibration: Southampton Workshop 2000. AB - OBJECTIVES: To identify the current state of knowledge, current uncertainties and future needs related to the diagnosis of disorders associated with the use of vibratory hand-held tools. METHOD: An international workshop was convened with invited experts, medical doctors, scientists and engineers familiar with hand transmitted vibration and the diagnosis of vascular, neurological and musculoskeletal disorders. This paper records the general conclusions from four panel discussions. RESULTS: For the most common vascular disorder (vibration induced white finger), the principal symptom and sign involves attacks of well demarcated finger blanching (Raynaud's phenomenon); low finger systolic blood pressure following cooling is indicative of vibration-induced white finger and zero finger systolic blood pressure can confirm an attack of Raynaud's phenomenon. For neurological disorders, some symptoms can exist without detectable signs and some signs can exist without symptoms; numbness and tingling are commonly reported but neurological changes may be present without these symptoms. The pathogenesis of musculoskeletal disorders in users of vibratory tools is not clear; symptoms may include pain that may not be associated with abnormal results in objective tests. For both neurological and musculoskeletal disorders, a thorough neuromuscular and skeletal examination is required; diagnosis must consider the work history and medical history, the results of physical examination and any objective tests in addition to other factors (e.g. age, smoking, alcohol, systemic disorders, medication and neurotoxic agents) that might have contributed to symptoms, signs and test results. CONCLUSIONS: While vibration-induced white finger is caused by vibration, some neurological and musculoskeletal disorders are the result of work with vibratory tools where the separate roles of vibration, repetitive movements, grip and push forces, non neutral postures and any other ergonomic stressors are often unclear. Such disorders may be more easily identified as being caused by the work rather than by exposure to hand-transmitted vibration per se. A person found to have developed disorders induced by either vibration or the work situation should not be returned to the same vibration exposure or work without any changes expected to lessen the risks. PMID- 11898869 TI - Measurement of manipulative dexterity in patients with hand-arm vibration syndrome. AB - OBJECTIVES: The purpose of the study was to assess quantitative measurements of the manipulative dexterity of patients suffering from hand-transmitted vibration disorder, and to investigate a possible link with impaired tactile perception in their hands. SUBJECTS AND METHODS: The subjects were 30 patients with hand-arm vibration syndrome, 30 office workers and 20 manual laborers of similar ages. Manipulative dexterity was assessed by measuring the performance time of buttoning-unbuttoning a work jacket with five buttons and of picking up and transferring 30 red beans, one by one, with the fingers from one plate to another. Patients were also examined on vibrotactile thresholds at 125 Hz as well as pain thresholds. RESULTS: The buttoning-unbuttoning time and the bean transfer time were significantly prolonged in the patients compared with the worker groups. The patients showed a close correlation between the buttoning-unbuttoning time and the bean transfer time. Performance times did not differ between patients with vibration-induced white finger and those without it. Those times also correlated with the vibrotactile and pain thresholds in the patients. When a cut-off value was set at 2 SDs from the mean performance times of the manual laborers, exceeding the cut-off value was found in 70% of the patients for the button-unbutton test and in 80% of those for the bean transfer test. All patients with moderately or severely increased vibrotactile and pain thresholds exceeded that value in performing the two tasks. CONCLUSION: The present measurements quantitatively showed impaired manipulation in patients with hand-arm vibration syndrome. Such difficulty with manipulation seems to be associated with the impaired tactile sensation in their hands. PMID- 11898870 TI - Repeatability of grip strength and dexterity tests and the effects of age and gender. AB - OBJECTIVES: Users of hand-held vibratory tools report reductions in grip strength and manual dexterity. This study quantified the test-retest repeatability of grip strength and manual dexterity tests, investigated effects of gender and age, and determined normative measures in different subject groups. METHODS: A total of 72 subjects in four groups (both genders and two age ranges) participated: men and women aged 18 to 25 years and 45 to 55 years. Grip strength was measured with a hand-held dynamometer, and dexterity measured with the Purdue pegboard. We assessed repeatability using one subject group (18 to 25-year-old men) who attended over three successive weeks. RESULTS: Repeated measures of grip strength were correlated for both hands and for each combination of weekly tests (P=0.01), and there were no significant changes in strength over weeks. Repeated measures of dexterity were correlated in both hands (P=0.01) for all test combinations, except between weeks 1 and 3 in the non-dominant hand (P=0.15). Further analysis suggested an improvement in dexterity, consistent with a practice effect. In both age groups, grip strength of the men was significantly greater than that of the women (P<0.01), but there were no gender differences in dexterity scores (P>0.1). There were no significant effects of age for either grip strength or dexterity (P>0.1). CONCLUSIONS: Both tests showed sufficient repeatability, with no age effect on either grip or manual dexterity (between approximately 20 to 55 years), although a decline in grip and dexterity is expected at greater ages. Dexterity scores were similar in both genders for the groups studied. Grip strength was greater in men. Occupational effects might exist for both tests, irrespective of any occupational disorder, and might be reflected in increases or decreases in grip strength and dexterity. PMID- 11898871 TI - National regulations for diagnostics in health surveillance, therapy and compensation of hand-transmitted vibration injury in Japan. AB - OBJECTIVES: During the period of technological innovation and rapid economic development, portable power tools were introduced on a large scale in Japan. Vibration disease from the operation of those tools and its prevention and therapy became urgent social problems in the 1970s. This paper aims to introduce national regulations in Japan for diagnostics in the health surveillance, certification, therapy and compensation of vibration disease and evaluates them in the present perspective. METHOD: Relevant laws, regulations and administrative directives were described in chronological order. Effect of those laws, regulations and directives were evaluated by statistics. RESULTS: Relevant regulations were established in 1947 and were revised in the 1960s and 1970s. According to those regulations, administrative directives were issued. Relevant vibration-disease statistics improved from the 1970s to 1990s. The annual ratio of workers examined was 95% to 100% in national forests (NFs), 47.3% in 1980 and 40.8% in 1990 in private industry (PI). The number of workers certified in NFs was 1,796 from 1971-1975, with a decrease to nine from 1991-1995, while in PI there were 9,783 from 1976-1980, decreasing to 2,331 from 1991-1995. However, in the construction industry the number increased again in the 1990s. The top four workers certified by the type of tool from 1994-1997 were operators of rock drills, chainsaws, pick hammers and concrete vibrators. The annual number of workers under treatment (at highest level) was 3,605 (1982; NFs) and 13,501 (1987; PI), with a decrease to 3,481 (1997; NFs) and 8,958 (1997; PI). Regulations for compensation covered 3,670 workers from 1965 to 1997 (NFs) and 22,723 from 1976 to 1997 (PI) in medical treatment benefits, and 189 (NFs) and 15,448 (PI) in disability benefits during the same term. CONCLUSION: The national regulations developed in Japan since 1965 for health surveillance, certification, therapy and compensation of hand-transmitted vibration disease have proven effective for prevention and compensation of vibration disease in many industries, but unsolved problems remain in the construction industry. PMID- 11898872 TI - Cold-stress tests involving finger skin temperature measurement for evaluation of vascular disorders in hand-arm vibration syndrome: review of the literature. AB - Cold-stress tests are used for evaluating vascular disorders in the hand-arm vibration syndrome, and the value of such tests based on finger skin temperature measurement has been investigated. However, there is a wide difference in the test conditions among countries and researchers. Standardization of the cold stress tests is currently under discussion within the International Organization for Standardization. We reviewed various aspects of the cold-stress tests involving finger skin temperature measurement, including water temperature, hand immersion time and other test conditions, and evaluated their diagnostic significance. Water temperature varied from 0 degrees C to 15 degrees C and hand immersion time varied from 0.5 min to 20 min. The cold-stress tests are associated with relatively severe suffering, thus, higher temperature of cold water and shorter time of immersion are desirable. To date, however, there has not been sufficient data indicating diagnostic value in a test method involving cold water at around 15 degrees C. Diagnostic value is also influenced by other test conditions, such as room temperature, season, use of ischemia during immersion. For standardization of the cold-stress test involving finger skin temperature measurement, these factors must be considered together with water temperature and immersion time. PMID- 11898873 TI - Finger systolic blood pressure indices for the diagnosis of vibration-induced white finger. AB - OBJECTIVES: To compare the accuracy of several finger systolic blood pressure indices (FSBPIs) for the diagnosis of cold-induced Raynaud's phenomenon in vibration-exposed worker groups with different prevalences of vibration-induced white finger (VWF). METHODS: The finger systolic blood pressure (FSBP) in a test finger at 10 degrees C as a percentage of the pressure at 30 degrees C, corrected for the change in systolic blood pressure in a reference finger [FSBPI(A)] or the arm [FSBPI(B)], was measured in 455 healthy controls and 874 workers exposed to hand-transmitted vibration (HTV). The following FSBPIs were also calculated: FSBPI(C), as the ratio between FSBP in the test finger at 10 degrees C and FSBP in the same finger at 30 degrees C; and FSBPI(D), as the ratio of FSBP to arm systolic blood pressure during finger cooling to 10 degrees C. The finding of zero systolic blood pressure, FSBP(0), in the cooled finger was taken as an objective sign of Raynaud's attack with complete closure of the digital arteries. RESULTS: On a group basis, all FSBPIs could discriminate between the controls and the HTV workers. In the vibration-exposed worker population, the FSBPIs were significantly lower in the subjects affected with VWF than in those without vasospastic symptoms. The lower normal limits of FSBPIs were derived from the results of the cold test in the controls and were found to vary from 50% to 60%. The FSBPI(A) showed the best sensitivity for the detection of cold-induced digital arterial hyper-responsiveness in both the total sample of HTV workers (sensitivity 87%) and most of the vibration-exposed groups with different prevalence of VWF (sensitivity from 79% to 100%). The results of the receiver operating characteristic analysis suggested a higher diagnostic accuracy of FSBPI(A) when compared with the global performance of the other FSBPIs. In the whole sample of HTV workers, the predictive value of a positive cold test varied from 73% [FSBPI(D)] to 89% [FSBP(0)] and that of a negative test ranged between 89% [FSBP(0)] and 97% [FSBPI(A)]. CONCLUSIONS: The findings of this investigation and clinical experience suggest that a discriminating threshold of FSBPI(A) <60% during finger cooling to 10 degrees C is an appropriate diagnostic criterion for the detection of abnormal cold response in the digital arteries of most vibration exposed workers with a true history of symptoms of finger whiteness at the medical interview. PMID- 11898874 TI - The clinical grading of Raynaud's phenomenon and vibration-induced white finger: relationship between finger blanching and difficulties in using the upper limb. AB - OBJECTIVES: To investigate the association between functional difficulties in using the upper limb and extent and frequency of finger blanching, and the merits of these markers in grading the severity of Raynaud's phenomenon (RP) and vibration-induced white finger (VWF). METHODS: A questionnaire was mailed to a randomly selected community sample of 22,194 working-aged adults. Information was collected on cold-induced finger blanching--including the extent and frequency of attacks in the past year, and on difficulty in using the upper limb in several everyday activities (e.g. doing up buttons, opening a tight screwtop jar, and pouring from a jug). Associations were examined by logistic regression with the resultant odds ratios converted into prevalence ratios (PRs) and 95% confidence intervals (95% CI). RESULTS: Among 12,907 respondents were 1,359 who reported finger blanching and provided details of its extent and frequency. Of these, 7.4% reported frequent attacks (50 or more over the year), and 12% reported extensive blanching (affecting nine or ten digits). After adjustment for potential confounders, subjects with finger blanching reported an excess of difficulties in using the limb. Thus, in men with blanching the PR for difficulty with buttons was 4.7 (95% CI 3.9-5.8), and that for pouring from a jug was 3.8 (3.0-4.9) in comparison with men who had never had blanching. Similar associations were found in women and in those men with exposure to hand-transmitted vibration. The risk of reporting difficulties increased markedly with frequency of blanching--up to four- or fivefold in those with 50 or more attacks in the past year compared with those who had none; but differences by extent were less marked, with PRs < or = 1.6 in those with nine to ten digits affected compared with one to two digits. CONCLUSIONS: RP and VWF are both associated with difficulties in using the upper limb in everyday tasks. Further investigation of potential reporting biases is warranted, but if the associations are causal, frequency of attacks influences impairment more than extent of disease. More account may need to be taken of frequency of blanching episodes in assessing and in compensating subjects with VWF. PMID- 11898875 TI - Use of questionnaire screening for vibration white finger in a high risk industrial population. AB - OBJECTIVES: To compare the results of questionnaire screening with those of objective testing in the identification of vibration white finger (VWF) in a high risk population. METHODS: Three groups of men (79 riveters, 52 healthy controls and 79 compensation claimants) were assessed for VWF by a detailed questionnaire covering occupational and general medical history and incorporating specific questions related to the presence or absence of relevant symptoms in their hands. Each then underwent provocative cold testing under controlled conditions using established protocols. The presence or absence of digital vasospasm following cooling was determined by finger systolic pressure measurements using laser Doppler flowmetry. The test protocols used have been evaluated in patients with a clear clinical diagnosis of non-occupational Raynaud's syndrome. RESULTS: 6.3% of the riveters and 83.5% of the claimants reported specific Raynaud's syndrome symptoms but 30.4% of the riveters and only 19% of the claimants tested positive for vasospasm after middle phalangeal cooling to 10 degrees C for 5 min. Using a more severe cooling protocol provoked vasospasm in 46.8% of the claimants. Lack of sensitivity or specificity of the objective testing could not explain the large discrepancies between the findings in the riveters and in the claimants. CONCLUSIONS: Questionnaire responses concerning VWF symptoms can be influenced by the context in which they are recorded. It is important to employ more objective methods in assessing all workers at risk of developing VWF. PMID- 11898876 TI - Normative data for vascular and neurological tests of the hand-arm vibration syndrome. AB - OBJECTIVES: To assist occupational health professionals to interpret the results of standardised tests for components of the hand-arm vibration syndrome by presenting data for healthy subjects and identifying the effects of some of the confounding variables. METHOD: Thermal thresholds, vibrotactile thresholds, the finger skin temperature (FST) response to cold provocation and percentage finger systolic blood pressures (%FSBP) were measured by standardised procedures. Normative data were obtained for healthy men of working age (17-62 years) during 237 experimental sessions encompassing ten different studies. Hot thermal thresholds and cold thermal thresholds were assessed independently with 38 subjects; 152 measurements of both hot and cold thresholds were made. Vibrotactile thresholds were measured at several locations on 81 subjects, giving a total of 216 measurements at 125 Hz and at 31.5 Hz. The FST response to cold provocation at 15 degrees C was monitored by thermocouples throughout a 2-min settling period, a 5-min immersion period and a 10-min recovery period. A total of 302 measurements was made on 70 subjects. The %FSBPs were measured in four test fingers and one reference finger by strain-gauge plethysmography. Measurements were made on 97 subjects. A total of 351 measurements was made at 15 degrees C, with 341 measurements at 10 degrees C. RESULTS: Normative data and some example normal limits are presented from the current data set and from data presented in other studies. Age was found to influence thermal thresholds, vibrotactile thresholds and the FST response to cold provocation; older subjects exhibited deteriorated vascular and neurological function. Room temperature was found to influence %FSBPs and the FST response to cold provocation; warmer environments resulted in improved vascular response to cold. Outdoor temperature had a small effect on the FST response to cold provocation and on the vibrotactile thresholds. Thermal thresholds showed some influence of smoking habits and of the FST measured prior to testing. For all four tests, any differences between measurement locations were small and there were no differences between left-handed and right-handed subjects. CONCLUSIONS: The current data can assist occupational health professionals to interpret the results of the standardised tests. Comparison with the current data is considered valid for men of working age. Age and room temperature should be recognised as being capable of causing changes in neurological and vascular function. PMID- 11898877 TI - Neurological diagnosis: aspects of bedside and electrodiagnostic examinations in relation to hand-arm vibration syndrome. AB - OBJECTIVES: The objective of this paper, was to direct attention to the diagnostic strategy and clinical approach necessary in the diagnosis of neuropathy in workers exposed to vibration. The purpose encompassed evaluation of selected aspects of bedside and electrodiagnostic examinations with respect to biological validity and the ability to distinguish between subjects with and without neuropathy. METHODS: The neurological examinations viewed were restricted to those applicable to the upper extremity and neck system. A MEDLINE search was performed through the clinical queries service of PubMed searching for the following terms: nerve-conduction, Tinel's test, Phalen's test, tendon reflex, two-point discrimination test, abduction external rotation test, and Spurling test. Retrieved articles were discussed both in relation to the test accuracy and the validity aspects of the tests. RESULTS: The evidence in support of the view that neurological tests can accurately distinguish between subjects with and without neuropathy specifically addressing hand-arm vibration syndrome was sparse. The initial number of diagnostic hypotheses could be reduced by progressively ruling out diseases based on negative results of highly sensitive tests. As the possible diagnostic alternatives become fewer, the use of positive results from highly specific tests are more effective. The information value of the various diagnostic tests is determined by the change in pre-test to post-test probability of target disorder, which depends on the prevalence of the disorder and the likelihood ratios of the tests. The review showed that target disease characteristics influence the test outcome as well as the choice of "gold standard" and the population domain of the studies. CONCLUSIONS: The selection of various bedside examinations and diagnostic electrophysiological tests should be dependent on the clinical context, the history and results from the successive diagnostic tests. PMID- 11898878 TI - Diagnostic aspects of vibration-induced white finger. AB - Vibration-induced white finger (VWF) is a secondary type of Raynaud's phenomenon (RP) caused by exposure to hand-arm vibration. The present review concerns the cold-provoked attack of RP in vasospastic VWF. It concentrates on the most common clinical and laboratory methods used to diagnose RP in vibration-exposed subjects. Some physiological aspects of the attack of RP are mentioned to elucidate the diagnostic principles of the tests. Anamnestic diagnostics by medical interviews and questionnaires as well as cold-provocation tests with detection of finger colour, finger systolic blood pressure (FSP), recovery time of finger skin temperature and recovery time of normal nail colour after nail compression are mentioned. The discriminative capacity and the reproducibility of the tests are discussed. Cold-provocation tests with detection of finger colour or zero FSP during cooling are recommended to be used if an attack of RP has to be registered for diagnostic or medico-legal purposes in individual cases. An abnormal reduction in FSP during cooling makes a history of RP very probable and is a suitable laboratory test for groups of subjects. Both recovery tests may be useful screening tests in field studies of vibration-exposed subject groups. PMID- 11898879 TI - Neurological diagnosis--aspects of quantitative sensory testing methodology in relation to hand-arm vibration syndrome. AB - OBJECTIVE: The objectives are to summarise the fundamental neurophysiological base for quantitative sensory testing (QST), and to discuss associated methodological and practical aspects necessary to consider with respect to applicability and reliability as a screening or diagnostic aid for vibration induced sensory neuropathy. RESULTS: QST is the use of psychophysical techniques to measure the intensity of stimuli needed to produce specific sensory perceptions. The physical components are graded stimuli presented to the skin, eye or ear. The psychological component is mental recognition of the physical stimulus. Sensory modalities are named after the subjective feelings elicited, i.e. touch, pressure, vibration, warmth, cold and thermal pain. Since an exposure to vibration may cause symptoms and signs of sensory neuropathy in the hand and arm, the use of QST as an aid for screening and diagnosis has gained an increasing interest. The "Stockholm Workshop" classification scale for sensorineural stages also requires QST. Further, QST has also spread into many other areas, such as in the screening and diagnosis of peripheral neuropathy or polyneuropathy induced by different types of illness, exposure to toxic substances, compression and nerve entrapment. CONCLUSION: QST is in general easy to perform, usually not associated with pain (except thermal pain), suitable for screening and can readily be conducted in the field. QST is, however, known to be susceptible to the effects of multiple covariates and test methodologies. It is thus important that the relative influence on test results from all significant covariates are identified, and to standardise test methodology accordingly before QST can become a reliable and useful tool for diagnostic and screening purposes in the field of vibration-induced sensory neuropathy. The sensitivity, specificity and reliability of different methods for QST for this type of disorder is still very much unknown. Lack of normative values, standardisation of methods and of a "gold standard" for the presence of sensory neuropathy are some reasons. PMID- 11898880 TI - Dependence of vibrotactile thresholds on the psychophysical measurement method. AB - OBJECTIVES: To investigate the dependence of vibrotactile thresholds on the psychophysical method used in the diagnosis of neurological dysfunction caused by exposure to hand-transmitted vibration. To compare thresholds obtained with (a) 'continuously variable' versus intermittent 'staircase' stimulation using 'yes no' responses, and (b) 'yes-no' and 'forced-choice' responses using intermittent staircase stimulation. METHODS: Vibrotactile thresholds were measured on 12 healthy men by three different psychophysical methods. All measurements were performed with the same vibrometer in which the vibratory stimulus was applied by a probe 6 mm in diameter that protruded through a hole of 10 mm diameter in a surround, controlling both the contact force and the push force. Four stimulus frequencies (16, 31.5, 63 and 125 Hz) were used to obtain responses from FAI and FAII mechanoreceptors. RESULTS: There was a 3 to 6 dB variation in threshold due to the psychophysical method: thresholds were lower with intermittent stimulation and thresholds obtained with the 'forced-choice' procedure were lower than those obtained with the 'yes-no' procedure. Alternative explanations of the findings were offered. CONCLUSIONS: The dependence of psychophysical measurement method on vibrotactile thresholds was partly due to influencing responses via mechanoreceptor systems. It was suggested that the psychophysical measurement method had a sufficiently large effect on vibrotactile thresholds for it to be taken into account when methods for the diagnosis of neurological disorders are standardised. PMID- 11898881 TI - A comparison of vibrotactile thresholds obtained using different diagnostic equipment: the effect of contact conditions. AB - OBJECTIVES: Vibrotactile thresholds on the fingers were compared using two alternative methods of controlling contact with a vibrating probe: control of the force of contact with the probe (force control) and control of skin indentation produced by the probe (indentation control). Both systems had the same control of push force on a static surround around the vibrating probe. METHOD: A group of 14 male subjects (aged 20-27 years) were tested at four frequencies (31.5, 63, 125, 250 Hz) in three separate sessions so as to quantify the repeatability of thresholds. Skin stiffness was also measured. RESULTS: Control of skin indentation gave more repeatable thresholds than control of probe force. There was a practice effect whereby thresholds became more consistent over sessions. There were no systematic correlations between thresholds and skin stiffness. CONCLUSIONS: Repeatable and similar vibrotactile thresholds can be obtained with two alternative methods having different contact conditions. Either method may assist the diagnosis of disorders associated with hand-transmitted vibration, but control of skin indentation has the advantage of greater simplicity and, in this study, greater repeatability. PMID- 11898882 TI - Thermotactile threshold testing for the evaluation of sensory nerve function in vibration-exposed patients and workers. AB - OBJECTIVES: The aim of the present study was to ascertain the usefulness of thermotactile testing under the measuring conditions clarified by our previous study in the evaluation of vibration-induced neuropathy. METHOD: Thermal (warm and cold) thresholds were examined on the index fingers of both hands of ten patients with hand-arm vibration syndrome, 36 workers exposed to hand-transmitted vibration, and ten healthy controls, using an aesthesiometer under the following measuring conditions: change rate of the applicator temperature was fixed at a rate of 1 degrees C/s; finger contact force on the applicator was kept at 0.5 N; and forearm was positioned on a support. Vibrotactile thresholds at 125 Hz and pain thresholds were also measured on the index fingers. RESULTS: Both warm and cold thresholds had more severely deteriorated in the patients than in the controls. Vibrotactile and pain thresholds were also increased in the patients. In the vibration-exposed workers, the neutral zone between the warm and cold threshold differed from in the controls. Thermotactile thresholds were likely to have deteriorated with advanced nerve impairment. Likelihood ratios of thermotactile testing were significantly high between the patients and the controls. For vibration-exposed workers with elevated vibrotactile and pain thresholds, the ratios were also significantly high, particularly in the neutral zone, though the accuracy tended to be lower than in the patients. Thermotactile thresholds appeared to be more closely correlated with pain thresholds than vibrotactile thresholds. CONCLUSIONS: Thermotactile threshold testing under the present measuring conditions can be useful for evaluating small sensory nerve fibre dysfunction in vibration-exposed subjects, and the neutral zone may be a sensitive indicator in vibration-exposed workers who may have slight or mild nerve impairments. PMID- 11898883 TI - Clinical assessment of musculoskeletal disorders in workers exposed to hand-arm vibration. AB - OBJECTIVES: To describe the clinical assessment of musculoskeletal disorders among vibration-exposed workers and to review the experimental and epidemiological studies of the effects of vibration on the musculoskeletal system of the upper limbs. METHODS: A total of 212 references in English was found in Pub Med for the years 1980-2000 that dealt with clinical assessment. Many of these references were reviews and few were original research dealing with test performance in diagnostic procedures. RESULTS: The reported effects on bone are osteoporosis and cysts in the hands. Experiments have shown injuries to muscle cells in animals and additional physiological loading of muscles in humans by vibration. Low-frequency vibration exposure of high magnitude was associated with osteoarthrosis in the elbow, wrist and acromioclavicular joint and symptoms in the elbow and shoulder. Impacts, jerks and blows with high-energy transfer to the hands at low frequency might have the potential to result in musculoskeletal disorders considering the general model for injuries. Furthermore, the observed associations with vibration exposure and musculoskeletal disorders might result from the strong dynamic and static joint loading and the repetitive hand-arm motions required in tasks where hand-held machines are used. The clinical assessment of musculoskeletal disorders in workers exposed to hand-arm vibration consists of the clinical and exposure history and evaluation of the physical and laboratory findings. Since most patients with musculoskeletal disorders who are exposed to vibration are also exposed to other ergonomic stressors, accommodation of the injured worker has to take the whole work system into account (task, technology, environment and organisation). CONCLUSIONS: The scientific evidence that vibration per se is a risk factor for musculoskeletal disorders is still weak although there is strong evidence that job tasks with vibrating machines are associated with musculoskeletal disorders. The clinical assessment of musculoskeletal disorders in exposed patients imposes special requirements. PMID- 11898884 TI - The rationale for aerosolized antibiotics. AB - In order for an antimicrobial agent to be effective, it must fulfill two requirements. First, the agent must reach the site of infection and remain in the vicinity for an adequate length of time. Second, it must bind to a target site and remain bound for a length of time sufficient to disrupt the life cycle of the cell. Once these requirements are met, the drug is able to exert its antimicrobial activity against the cell. In an effort to better understand and predict the killing activity of antibiotics, we have attempted to develop parameters that describe the accumulation and diffusion of drug to and from body sites (pharmacokinetics) and quantify how much of a compound is needed at the site of infection to yield the desired effect (minimum inhibitory concentration). Furthermore, integration of these parameters allows us to evaluate host, drug, and microbial factors and formulate criteria to assess and predict drug activity in patients (pharmacodynamics). Knowledge and application of pharmacodynamic principles can assist clinicians in optimizing antimicrobial therapy by allowing them to maximize the antimicrobial activity of an agent while minimizing patient exposure and thus reducing the likelihood of toxicity. PMID- 11898885 TI - Pharmaceutical considerations in aerosol drug delivery. AB - Aerosolized drug delivery has been used for over 50 years, but its quality and scope continue to increase. Three factors affect this form of drug delivery: the nebulizer, the compressor, and the actual drug preparation. As technology and knowledge improve, this delivery system also improves. Patients with cystic fibrosis, in particular, can benefit significantly from this form of therapy. PMID- 11898886 TI - Treatment options for cystic fibrosis: case study and panel discussion. PMID- 11898887 TI - Lessons learned from the september 11th World Trade Center disaster: pharmacy preparedness and participation in an international medical and surgical response team. PMID- 11898888 TI - Health literacy: a review. AB - Illiteracy has become an increasingly important problem, especially as it relates to health care. A national survey found that almost half of the adult population has deficiencies in reading or computation skills. Literacy is defined as the basic ability to read and speak English, whereas functional health literacy is the ability to read, understand, and act on health information. Up to 48% of English-speaking patients do not have adequate functional health literacy. The consequences of inadequate health literacy include poorer health status, lack of knowledge about medical care and medical conditions, decreased comprehension of medical information, lack of understanding and use of preventive services, poorer self-reported health, poorer compliance rates, increased hospitalizations, and increased health care costs. The medical community must acknowledge this issue and develop strategies to ensure that patients receive assistance in overcoming the barriers that limit their ability to function adequately in the health care environment. PMID- 11898889 TI - Association between acetaminophen or nonsteroidal antiinflammatory drugs and risk of developing ovarian, breast, or colon cancer. AB - STUDY OBJECTIVE: To explore the association between exposure to acetaminophen (paracetamol) or nonsteroidal antiinflammatory drugs (NSAIDs) and risk of developing ovarian, breast, or colon cancer. DESIGN: Retrospective case-control study SETTING: General practice offices. SUBJECTS: Four hundred eighty-three women with ovarian cancer and 1877 women matched for age, years of medical history in computer record, general practice attended, and calendar time; 3706 women with breast cancer and 14,155 matched control subjects; and 635 women with colon cancer and 2434 matched control subjects. INTERVENTION: United Kingdom based General Practice Research Database was searched for women aged 50-89 years with a first-time diagnosis of ovarian, breast, or colon cancer and for matched controls to assess prescription analgesic exposure. MEASUREMENTS AND MAIN RESULTS: Regular acetaminophen exposure (> or = 30 prescriptions) was associated with a slightly decreased risk of developing breast (odds ratio [OR] 0.8, 95% confidence interval [CI] 0.7-1.0) but not ovarian (OR 1.0, 95% CI 0.6-1.5) or colon (OR 1.0, 95% Cl 0.7-1.4) cancer. Regular NSAID exposure was associated with a reduced risk of colon (OR 0.5, 95% CI 0.3-0.9) but not ovarian or breast cancer. CONCLUSION: We found no evidence for a decreased ovarian cancer risk for women with regular acetaminophen or NSAID exposure. PMID- 11898890 TI - Strategies to reduce toxicities and improve outcomes in renal transplant recipients. AB - Ongoing improvements in immunosuppression and posttransplantation care have dramatically improved patient and graft outcomes after transplantation. The frequency of graft loss due to acute rejection has declined considerably as a result of the availability of a variety of more potent immunosuppressive agents and probably also because of refined clinical care and follow-up. Complications of long-term administration of corticosteroids (steroids) and calcineurin inhibitors, however, have become increasingly apparent as patients live longer with their transplant, and attention is shifting to long-term issues. Use of both steroids and calcineurin inhibitors is associated with metabolic toxicities such as hypertension, hyperlipidemia, diabetes, bone loss, and cataracts. These contribute to posttransplantation morbidity and may negatively affect patient and allograft survival. A variety of troublesome cosmetic side effects, such as hirsutism, gingival hyperplasia, alopecia, obesity, and cushingoid appearance, also are associated with these drugs. These effects can detract from patient self esteem and compliance with the immunosuppressive regimen. In the past 2 decades, the introduction of second-generation immunosuppressive drugs, such as tacrolimus, mycophenolate mofetil, sirolimus, and anti-interleukin-2 receptor monoclonal antibodies, has provided some alternatives to classic immunosuppressant choices. Patients experiencing undesirable adverse events now can be converted to another immunosuppressive regimen that ultimately will improve graft and patient survival rates and improve quality of life after transplantation. PMID- 11898891 TI - Strategies for optimizing antiepileptic drug therapy in elderly people. AB - The elderly take more antiepileptic drugs (AEDs) than all other adults. This extensive use directly correlates with an increased prevalence of epilepsy in a growing population of older people, as well as other neuropsychiatric conditions such as neuropathic pain and behavioral disorders associated with dementia and for which AEDs are administered. The agents account for nearly 10% of all adverse drug reactions in the elderly and are the fourth leading cause of adverse drug reactions in nursing home residents. Numerous factors associated with advanced age contribute to the high frequency of untoward drug effects in this population; however, strategies are available to ensure optimal outcomes. PMID- 11898892 TI - Clinical outcomes of critical illness polyneuropathy. AB - It is often difficult to isolate the origin of acute weakness in the critically ill population because of multiple etiologies. Aminoglycosides, corticosteroids, and neuromuscular blockers frequently are implicated as the source of acute weakness. Recently, critical illness polyneuropathy (CIP), a syndrome of unknown etiology, was added to the differential diagnosis. The frequency of CIP is approximately 70% in patients with sepsis. Early studies of CIP, which were mostly retrospective, underestimated its frequency due to the complexity of the diagnosis and unfamiliarity with the syndrome. Prospective studies have explored the causality and clinical outcomes of CIP Clinical outcomes of patients with CIP include difficulty weaning from mechanical ventilation, increased length of stay, prolonged recovery, and an overall mortality rate of 26-71%. The association of CIP with sepsis, multiorgan failure, and drugs is still unclear. PMID- 11898894 TI - Gastrointestinal bleeds associated with rofecoxib. AB - Data suggest that cyclooxygenase (COX)-2 inhibitors are safer for the gastrointestinal tract than traditional nonsteroidal antiinflammatory drugs (NSAIDs). Rofecoxib, a COX-2-specific NSAID, does not inhibit the COX-1 enzyme, thereby decreasing the potential for gastrointestinal-related adverse effects. Patients who are at an increased risk for NSAID-related gastrointestinal bleeding are therefore appropriate candidates for COX-2 inhibitors. Although the agents provide benefits for many patients, gastrointestinal-related side effects may occur, and caution should be practiced when prescribing COX-2 inhibitors. We report two patients who were admitted to the hospital with diagnoses of gastrointestinal bleeds while they were taking rofecoxib. PMID- 11898893 TI - Potential anaphylactic shock with abciximab readministration. AB - A 46-year-old woman developed an anaphylactic reaction during percutaneous coronary intervention after she was pretreated with prednisone and diphenhydramine for a known allergy to iodine. She developed pruritus, edema, and nausea, which were followed by bradycardia and shock, minutes after administration of a bolus and standard-dose infusion of abciximab. The reaction was treated successfully with epinephrine, methoxamine, hydrocortisone, atropine, furosemide, sodium bicarbonate, diphenhydramine, and ranitidine. PMID- 11898895 TI - Intravenous cyclosporine-rifampin interaction in a pediatric bone marrow transplant recipient. AB - A 10-year-old girl with chronic myelogenous leukemia began receiving cyclosporine the day before bone marrow transplant surgery Three days after the transplant, she developed fever and neutropenia due to a Staphylococcus aureus bacteremia. Despite treatment with various antibiotics, the patient's fever persisted over the next 4 days. Intravenous rifampin was added to her antibiotic regimen of piperacillin, tobramycin, cloxacillin, and amphotericin. On day 12, the patient's blood cultures were negative and her fever had resolved; rifampin was discontinued. On day 16, the patient engrafted; she subsequently developed a grade II graft-versus-host disease of the skin and gastrointestinal tract, which responded to methylprednisolone. Her cyclosporine blood levels, which had been subtherapeutic since day 5 despite increasing intravenous dosages, were within the therapeutic range on day 21, and she was discharged 12 days later. To our knowledge, this is the first documented case of an intravenous cyclosporine rifampin interaction that resulted in subtherapeutic cyclosporine concentrations in a child receiving a bone marrow transplant who subsequently developed acute graft-versus-host disease. PMID- 11898896 TI - Possible mesalamine-induced pericarditis: case report and literature review. AB - Pericarditis should be considered in any patient complaining of chest pain and/or dyspnea who is taking a product that contains mesalamine or sulfasalazine. A 41 year-old woman was taking mesalamine 800 mg 3 times/day for 3 weeks before hospital admission. She complained of sharp, pleuritic chest pain that radiated down both arms and increased in intensity when lying down. She was diagnosed with pericarditis based on clinical presentation and electrocardiogram findings. Differential diagnoses for myocardial infarction, systemic lupus erythematosus, and viral or bacterial causes were ruled out based on subjective and objective data. Mesalamine-induced pericarditis was considered on hospital day 2, and the drug was discontinued at discharge on day 3. Clinicians should be aware of this potential drug-related complication, as the relationship between mesalamine or sulfasalazine and pericarditis has been reported rarely in the literature. PMID- 11898897 TI - Levofloxacin treatment failure in a patient with fluoroquinolone-resistant Streptococcus pneumoniae pneumonia. AB - The frequency of fluoroquinolone-resistant Streptococcus pneumoniae has increased as fluoroquinolone administration for treatment of respiratory tract infections has increased. Levofloxacin treatment failed in a patient who had pneumococcal pneumonia and had received three previous courses of levofloxacin therapy. Susceptibility testing revealed high-level resistance to levofloxacin (minimum inhibitory concentration [MIC] > 32 microg/ml), and cross-resistance to moxifloxacin (MIC 4 microg/ml), trovafloxacin (6 microg/ml), and gatifloxacin (12 microg/ml). Sequencing of the quinolone-resistance determining region revealed a mutation of serine-81 to phenylalanine (Ser81-->Phe) in the gyrA region of DNA gyrase and a Ser79-->Phe mutation in the parC region of topoisomerase IV The patient was treated successfully with intravenous ceftriaxone followed by oral cefprozil. Clinicians must be aware of local resistance patterns and the potential for fluoroquinolone treatment failures in patients with infections caused by S. pneumoniae. PMID- 11898898 TI - Amlodipine PREVENTS angina, not atherosclerosis. PMID- 11898899 TI - Temperature measurement for energy-efficient ablation by thermal radiation with a microsecond time constant from the corneal surface during ArF excimer laser ablation. AB - Measurement of the temperature of the corneal surface during photorefractive keratectomy (PRK) is thought to be useful for monitoring the corneal ablation process, since the photothermal process has been proposed as the major mechanism of ArF excimer laser ablation. For temperature measurement, we measured thermal radiation from the corneal surface during ArF excimer laser ablation using a mercury-cadmium-telluride detector with a 1-micros time constant. To investigate the effects of temperature on ablation depth, the ablation depth of the cornea was measured by microscopy. When corneal ablation was initiated at the fluence of 65 mJ/cm2, the corneal surface temperature rose to 60-70 degrees C. The energy required for a unit-depth ablation (degrees C/microm) was lowest at 120 micro C. Monitoring of transient temperature during PRK provides important information on energy-efficient ablation, which may enable rapid and safe corneal incisions. PMID- 11898900 TI - Development of a subject-standing-type cone-beam computed tomography for chest and orthopedic imaging. AB - A subject-standing-type cone-beam computed tomography (CT) with high spatial resolution has been developed as a new three-dimensional imaging modality for subjects standing or sitting naturally on a turntable. A 16-in. X-ray image intensifier and charge coupled device camera acquires a 12-bit 5122-pixel projection at 60 f/s and the rotation period is 4.8 or 9.6 s for 288 or 576 projections, respectively. To reduce image noise, the system controls the X-ray pulse duration and iris-opening area through real-time analysis of the projection image. To improve CT accuracy and eliminate artifacts, the veiling glare of the image intensifier and scattered X-rays are corrected. Human chest and orthopedic studies with about 50 patients were conducted. Three-dimensional images with a spherical field of view with a diameter of 21-25 cm, 0.4- to 0.5-mm voxels and a 512(3) matrix were obtained. In coronal, sagittal and volume rendering images, the surface of arthrosis was visualized smoothly with a resolution higher than that of conventional CT. In the case of gonarthrosis, narrowing of the clearance at the surface of arthrosis was visualized clearly under body-weight burdening, which would be difficult if the subject was lying down. PMID- 11898901 TI - Blink artifact elimination in electroencephalographic records based on discrete cosine transform domain modeling. AB - A method for eliminating blink artifacts that contaminate electroencephalographic (EEG) records is introduced. The proposed method enables a direct estimation of blink artifacts without the use of the simultaneously recorded electroculargraphic (EOG) signal. The estimation of blink artifacts in the EEG was achieved by modeling in the discrete cosine transform (DCT) domain. The DCT coefficients of the blink artifact were sufficiently represented by a system transfer function with a low model order of 4. Two cases were investigated: one was blink artifact elimination for the background EEG and the other was for the somatosensory evoked potentials. The effectiveness of the proposed method was demonstrated in an experimental study based on actual EEG data of 11 healthy subjects. The simplicity and effectiveness of the proposed method suggest that blink artifact elimination using DCT domain modeling can possibly be applied in various areas of neuroscience for improving EEG quality. PMID- 11898902 TI - Indirect arterial blood pressure measurement at the wrist using a pad-type square cuff and volume-oscillometric method. AB - Using theoretical and experimental approaches, we examined whether blood pressure at the wrist can be accurately measured by a volume-oscillometric method using a small pad-type square cuff placed above the radial artery (RA). Nuclear magnetic resonance imaging of the wrist allowed us to determine the geometry of two dimensional (2-D) finite-element models. Finite-element method (FEM) analysis predicted that the pressure transmission ratio (calculated tissue pressure over externally applied pressure; square cuff to RA) was 98.8% for a cuff with a bladder sidelength 0.25 times the wrist diameter placed on the skin surface between the tendons of the brachioradialis muscle (Ta) and the flexor capri radialis muscle (Tb) and over the site (L) at which RA crosses the most protuberant spot on the volar aspect of the distal end of the radius. In addition, FEM analysis using a 3-D finite-element model (constructed by extending the 2-D finite-element model at site L in the longitudinal direction) showed that for all square cuffs with bladder sidelengths greater than or equal to 0.25 times the wrist diameter, the external pressure was transmitted almost completely to RA beneath the cuff center. Moreover, when the bladder sidelength was 0.44 times the wrist diameter, but not 0.29 times the wrist diameter, the mean blood pressure measured at site L in human subjects was similar to that measured at the upper arm. Taken together, the theoretical and the experimental results suggest that (i) blood pressure at the wrist can be measured accurately using a pad-type square cuff placed on the skin surface between Ta and Tb at site L, and (ii) the minimum bladder sidelength for accurate readings is somewhere between one-third and one-half of the wrist diameter. PMID- 11898903 TI - Analysis of pathological tremors using the autoregression model. AB - The usefulness of analysis of acceleration data using an autoregression model (AR) for differential diagnosis of Parkinson's disease and other diseases with tremors was investigated. The order of the AR model used in this study was 7, in accordance with Akaike's final prediction error criterion. The subjects included 19 patients with Parkinson's disease; 21 patients with essential tremor, which mainly appears in old people, as well as Parkinson's disease; and 13 healthy old people as a control group. The results of analysis of acceleration data showed that the first prediction coefficient, just as the main tremor frequency, was a useful parameter for differentiating patients in the Parkinson's disease patient group and essential tremor patient group. The seventh prediction coefficient was found to be a useful parameter for distinguishing pathological tremors observed in Parkinson's disease and essential tremor disease from physiological tremors observed in healthy people. Although the usefulness of other prediction coefficients for differential diagnosis of Parkinson's disease and other diseases with tremors has not yet been clarified, the results of this study showed that information obtained from AR model parameters in addition to information on main tremor frequency is useful for the diagnosis of Parkinson's disease. PMID- 11898904 TI - Learning lessons. PMID- 11898905 TI - Towards a sociology of CAM and nursing. AB - Over recent years a sporadic, but not insignificant, sociology of complementary and alternative medicine (CAM) has begun to emerge. However, to date, the systematic sociological study of the apparent affinity between CAM, nursing as a profession and its practitioners has been absent from it. In this paper we argue for the need for a rigorous sociology of nursing and CAM and set out a provisional framework through which this might be operationalized. Three broad themes, as well as cross-cutting issues, are outlined. The way in which the challenges of CAM are mediated at the level of the individual and the profession are pivotal to analysis. An understanding of this mediation is crucial both as a means of extending knowledge, and as a means of engaging with complex issues such as the role of evidence, and the equity of provision, that are likely to accompany any extension of CAM mainstreaming. PMID- 11898906 TI - Establishment of a nurse-run acupuncture treatment for hayfever. By Mary Anne Cochrane [corrected]. AB - This interview describes how the need to set up an acupuncture clinic for the treatment of hayfever, was identified in a large teaching practice in south-east London. It then details the preliminary consultations for establishing the service, while taking into consideration the UKCC (1992a,b, 2000) guidelines, especially in relation to nurses' accountability and recognition of boundaries. It examines the issues of training and consultation time, as well as costs to the practice, and evaluates the benefits and costs of treatment to the patient. It also identifies some of the potential problems that may be encountered in providing acupuncture. The summary includes some guidance to others considering implementation of a similar service. PMID- 11898907 TI - Prayer in your practice. AB - This article is intended for anyone interested in introducing prayer into his or her practice. It outlines the reasons for using prayer and addresses some of the objections put forward by certain professionals.The paper then describesThe Prayer Wheel, a practical non-denominational way to pray and provides instructions on how to present it as an adjunct in health care. PMID- 11898908 TI - Research methodology for studies of prayer and distant healing. AB - The double-blind randomized clinical trial is the gold standard for trials of prayer and distant healing. Adequate blinding and randomization procedures should be followed and documented. The intervention must be well defined (include frequency, amount of time and training and/or experience level of healers). Subjects should have risks and benefits of study participation explained to them and sign informed consent before enrollment. Populations should be homogeneous. Consider stratification for smaller samples. Baseline information, including psychological status, beliefs about prayer and healing and other sources of prayer and healing, should be collected from subjects in clinical trials. This should be examined as part of the final data analysis for contribution to outcomes. Objectively measurable outcomes with adequate variability should be chosen. Subject study participation activities such as clinical interviews, traveling to special sites. journaling or meditation should be minimized to avoid washing out a small effect. In clinical trials subjects should be asked if they believed they were in the treatment group and this information should be entered as a co-variate for data analysis. Healers/prayers should be treated in a collegial and respectful way. Their healing efforts (time. location, method) should be documented in a log and they should be periodically contacted and encouraged by experimenters if the study' is taking place over an extended period of time. Observational and outcomes research can add an important dimension to healing research. Qualitative studies may also make an important contribution and help guide development of future controlled trials. PMID- 11898909 TI - Exploring attitudes toward, and knowledge of, homeopathy and CAM through focus groups. AB - Five focus groups (five female non-users; five male non-users; seven males, both users and non-users; seven female users; six male and female users) were conducted to get an idea of lay people's knowledge and attitudes to CAM. In each group, run by the same experienced moderator, various topics were systematically explored: knowledge of CAM treatments; attitudes towards CAM; personal experience of CAM; suggestions for bringing CAM into wider use. Results showed participants had self-confessed ignorance about homeopathy and also that their ideas and beliefs were not internally coherent. Suggestions about using this methodology are considered. PMID- 11898910 TI - Ernst E 2001 what's wrong with bias? PMID- 11898911 TI - Crossing cultures--images of East and West. AB - This article explores the challenges of working cross-culturally as a facilitator and teacher of imagework - an interactive imagery and personal development process. It discusses the use of imagery in different contexts and highlights the cultural background of Japan and the psychological conditions into which imagework was introduced. This unique approach demonstrates how the unconscious, through the symbolic language of images, expresses more about our humanity than it does about our cultural diversity. PMID- 11898912 TI - B cells underpin lupus immunopathology. PMID- 11898913 TI - Fluctuations in levels of antiphospholipid antibodies and increased coagulation activation markers in normal and heparin-treated antiphospholipid syndrome pregnancies. AB - Antiphospholipid antibodies (aPL) are associated with an increased risk of thrombosis and recurrent miscarriage. We assessed levels of coagulation activation markers and aPL during normal pregnancy and in women with the antiphospholipid syndrome (aPS). Fluctuations in aPL levels were observed in all patients with aPS. No particular pattern of antibody positivity, or fluctuation in aPL level, was associated with poor pregnancy outcome. A significant increase was observed in levels of factor Xlla (FXIIa; P < 0.001), factor VIIa (FVIIa, P < 0.001), thrombin antithrombin complexes (TAT; P < 0.001), prothrombin fragment F1.2 (F1.2; P < 0.001) and D-dimer (DD; P < 0.05) during normal pregnancy. Factor VIIa, TAT, F1.2 and DD increased significantly before 20 weeks gestation, while a statistically significant increase in FXIIa levels was first detected between weeks 20 and 30 of gestation. In pregnant women with aPS, increases in FXIIa were similar to those in normal pregnancy, but increased FVIIa levels were not observed until after 30 weeks gestation. Similar to normal pregnancy, increased levels of TAT and F1.2 were detected in aPS pregnancies before 20 weeks gestation, but increased DD were not observed until after week 20. Surprisingly, women with aPS receiving low molecular weight heparin prophylaxis had significantly higher (P = 0.02) levels of TAT (median 8.6; interquartile range (IQR) 6.5-20.8) between weeks 20 and 30 of gestation compared to the normal pregnant population (median 5.9; IQR 4.7-7.9), thus indicating increased thrombin generation in women with aPS in mid-pregnancy. PMID- 11898914 TI - Vascular endothelial growth factor plasma levels in patients with systemic lupus erythematosus and primary antiphospholipid syndrome. AB - The objective of this study was to assess the possible role of vascular endothelial growth factor (VEGF) in the pathogenesis of systemic lupus erythematosus (SLE) and primary antiphospholipid syndrome (PAPS). We studied 28 patients with SLE, 10 patients with PAPS, and 24 healthy controls. VEGF plasma levels were measured by ELISA. Immunolocalization of VEGF was done in renal tissue from SLE patients and cadaveric controls. Our results showed that VEGF plasma levels were increased in SLE patients compared with PAPS and controls. The correlation between clinical manifestations and VEGF levels revealed that SLE patients with renal failure had significantly increased plasma VEGF levels (134.1 + 91.0 pg/ml) compared with SLE patients with normal renal function (42.9 + 19.0 pg/ml), PAPS patients (41.9 + 26.6 pg/ml), and controls (36.2 + 27.0 pg/ml; P < 0.01). Immunostaining showed a strong expression of VEGF in SLE renal tissue samples. Our preliminary results indicate that VEGF is increased in plasma from patients with lupus nephritis and a moderate degree of renal failure and is overexpressed in renal tissue from these patients. PMID- 11898915 TI - Purified protein derivative reaction in systemic lupus erythematosus patients. Indirect study of cellular immunity. AB - Cutaneous anergy in SLE patients results from disease activity and/or immunosuppressive treatment (IT). The aim of this study was to evaluate purified protein derivative (PPD) reaction in SLE patients. A total of 145 patients and 20 controls were studied. Five units of PPD were applied on day 0, and skin reaction was measured after 3 (PPD1) and 6 (PPD2) days. A booster was applied (day 14), and the reaction was measured after 3 (PPD3) and 6 (PPD4) days. Non-parametric ANOVA test and unpaired Student's t-test were performed. Forty patients (group I) were inactive (MexSLEDAI < 3), receiving no IT (at least 3 months previous to the PPD test); 39 (group II) were inactive receiving IT; 24 (group III) were active without IT, and 42 (group IV) were active with IT. Active patients had lower PPD1 (group III, 1.4 +/- 0.9; group IV, 0.6 +/- 0.5) than inactive patients (group I, 8.4 +/- 2.3; group II, 5.1 +/- 1.9) and than controls (9.4 +/- 3; P < or = 0.001). Group IV had lower delayed response (PPD2 = 0.3 +/- 0.3) than inactive groups (group I, 2.6 +/- 0.9; group II, 3.1 +/- 0.8) and than controls (7.9 +/- 2.5; P < or = 0.001). Group III had lower delayed reaction (PPD2 = 1.2 +/- 0.8) than controls (P < or = 0.001). Active SLE patients, receiving or not receiving IT, had lower skin response to PPD than inactive patients and controls. PMID- 11898917 TI - Recent advances in antiphospholipid antibodies and antiphospholipid syndromes in pediatric populations. AB - In recent years, antiphospholipid antibodies (aPL) and their associated clinical features have been recognized increasingly in various pediatric autoimmune and non-autoimmune diseases. Pathogenic mechanisms involved in pediatric antiphospholipid syndrome (APS) appear to be the same as in adults. However, since pediatric patients do not have prothrombotic risk factors present in adults, there clearly are differences in the spectrum of clinical findings. The frequency of aPL-related thrombotic events is generally low in pediatric populations. On the other hand, various commonly acquired infections are likely to be responsible for higher percentage of non-pathogenic and transient aPL in childhood. Such points have to be considered in clinical judgment of elevated aPL in children. In this review we summarize the recent data on the prevalence and clinical significance of aPL in neonates, children and adolescents. PMID- 11898916 TI - Association of vitamin D receptor gene BsmI polymorphisms in Chinese patients with systemic lupus erythematosus. AB - The purpose of this study was to evaluate whether vitamin D receptor (VDR) genes BsmI polymorphisms were markers for susceptibility to or severity of systemic lupus erythematosus (SLE) in Chinese patients in Taiwan. The study included 47 Chinese patients with SLE. In addition, 90 unrelated, healthy individuals living in central Taiwan served as control subjects. Each polymorphism was detected as a result of polymerase chain reaction (PCR)-based restriction analysis. A PCR product length was determined to be 580bp (BB) whereas two fragments of 405 and 175bp were determined to be excisable lengths (bb) by BsmI endonuclease. The relationship between Bsm polymorphisms and clinical manifestations of SLE was evaluated. We found that BB was significantly more common and bb less common in SLE than in control group (chi2 = 54.2, P < 0.0001). In addition, the frequency of B allele was also significantly more common in patients with SLE than in the healthy control subjects (chi2 = 38.7, P < 0.0001), giving an odds ratio of 7.14 (95% confidence interval 3.53-14.4). In the SLE patients, we did not detect any associations of VDR genotype with the clinical, laboratory profiles, or lupus nephritis (chi2 = 2.34, P = 0.3). This study indicated an increased distribution of VDR BB genotype and B allelic frequencies in the Chinese SLE patients in Taiwan. However, there were no associations between the frequency of VDR allelic variations and clinical manifestations, laboratory profiles, or lupus nephritis. PMID- 11898918 TI - Conservation of FcepsilonRI gamma chain coding region in normals and in SLE patients. AB - High-frequency single nucleotide polymorphism (SNP) alleles are useful in mapping genes responsible for disease susceptibility. Functionally, Fcgamma receptors (FcgammaR) have been implicated in autoimmune disease, and the gene encoding the signaling element for several FcgammaR, Fc-epsilon-receptor gamma-chain (FcepsilonRIgamma), has several SNPs in the immunoreceptor tyrosine activation motif (ITAM) recorded in GenBank. Direct sequencing of the FcepsilonRIgamma coding region found potentially polymorphic sites in the 5'-->3' direction in control donors, which were not confirmed in the reverse direction (n = 66), and further exploration of 80 SLE patients revealed no non-synonymous SNPs. One normal donor was heterozygous for a non-synonymous SNP at nt 38 which changed the fifth codon from valine (GTG) to methionine (ATG). Although the EST databases suggest candidate SNPs, insertions and deletions, these appear to be artifacts, most probably due to secondary structure. The coding region of FcepsilonRIgamma shows a remarkable absence of nucleotide diversity. Either as yet unidentified regulatory elements of FcepsilonRIgamma or other genes in the region of human chromosome 1q23 are likely to be systemic lupus erythematosus disease susceptibility and severity genes. PMID- 11898919 TI - Reporting consistency in systemic lupus erythematosus patients: how reliable are patient histories? AB - Systemic lupus erythematosus (SLE) patients are frequently seen by multiple physicians and at multiple times. In each instance, most of the information important in clinical decision-making is gathered from the patient. There are no studies looking at reporting consistency of certain aspects of SLE patients' history. We studied this by administering the same nine-item questionnaire 4 months apart to the same cohort of SLE patients. In addition, a retrospective chart review was done to correlate the information obtained by prospective questioning and with that on the charts. Our results showed overall good consistency in the areas of general medical history, SLE-specific history and social history. The information gathered by the chart review, in general, went in parallel with that obtained by prospective questioning. This was also true for the poor correlation observed in the family history questions. Many studies and databases depend rather heavily on patient reporting and the quality of this information is usually not substantiated. Our study suggests that, even though SLE patients are generally consistent reporters of certain aspects of their histories, family history information provided is frequently not consistent with previous reporting. PMID- 11898920 TI - Reactivation of systemic lupus erythematosus in a dialysis patient after tuberculous peritonitis. AB - The disease activity of patients suffering from lupus nephritis usually becomes quiescent after the onset of end stage renal failure. Reactivation of lupus activity, especially after a long period of dialysis, is uncommon. Factors that might trigger off lupus reactivation after dialysis have not been well defined. We report a case of a 43-year-old Chinese woman on long-term peritoneal dialysis, who developed lupus reactivation with cerebral involvement 2 weeks after she was diagnosed to have tuberculous peritonitis. The close temporal relationship between the tuberculous peritonitis and the lupus reactivation raise the possibility that the tuberculous infection might have triggered off the lupus reactivation. PMID- 11898921 TI - Sustained normalization of cerebral blood-flow after iloprost therapy in a patient with neuropsychiatric systemic lupus erythematosus. AB - We report the case of a 30-year-old caucasian woman affected by SLE who developed neurological symptoms (prosopagnosia and visual-spatial agnosia) after nine years of disease. Brain MRI showed no abnormalities while a brain SPECT scan showed diffuse uptake defects and hypoperfusion areas in the right and left frontal parietal regions. At that time the patient was on hydroxychloroquine (400 mg/day) and oral prednisolone (0.5 mg/kg/day) as maintenance therapy. One year later the patient showed worsening of Raynaud's phenomenon with digital dystrophic lesions and was therefore treated with an intravenous infusion of Iloprost (1.5 ng/kg/min per 6h/day for 10 days consecutively), while baseline treatment remained unchanged. One month later the patient showed a dramatic improvement in her cognitive function and subsequent SPECT scans showed the gradual disappearance of perfusion abnormalities. This first report of Iloprost treatment in CNS lupus suggests the potential therapeutic usefulness of this drug in patients with SLE and functional CNS involvement. PMID- 11898922 TI - Lung cavitation in primary antiphospholipid syndrome. AB - Pulmonary complications of primary antiphospholipid syndrome are common and diverse, with thromboembolic events counting as the most frequent manifestation. We present the case of a female patient with a diagnosis of primary antiphospholipid syndrome, pulmonary thromboembolism and infarction followed by lung cavitation. PMID- 11898923 TI - Human immunodeficiency virus infection and systemic lupus erythematosus. An unusual case and a review of the literature. AB - Systemic lupus erythematosus (SLE) and infection with the human immunodeficiency virus (HIV) are rarely seen in the same patient. Both diseases share clinical and serological features and the differential diagnosis is difficult, with renal manifestations being of special interest. To date, 29 cases of association between the two diseases have been reported, but the diagnosis was simultaneous in just two of these and only 18 fulfilled the ARA criteria for the diagnosis of SLE. Most patients experienced an improvement in their SLE after development of their HIV associated immunosuppression and a reactivation of lupus manifestations has also been noted after immunological recovery secondary to antiretroviral therapy. We present the case of a woman in whom HIV and SLE with renal involvement were diagnosed simultaneously. PMID- 11898924 TI - Chemoadsorbtion apheresis: a useful therapy in lupus nephritis. PMID- 11898925 TI - Barriers to dietary control among pregnant women with phenylketonuria--United States, 1998-2000. AB - Newborns in the United States are screened for phenylketonuria (PKU), a metabolic disorder that when left untreated is characterized by elevated blood phenylalanine (phe) levels and severe mental retardation (MR). An estimated 3,000 4,000 U.S.-born women of reproductive age with PKU have not gotten severe MR because as newborns their diets were severely restricted in the intake of protein containing foods and were supplemented with medical foods (e.g., amino acid modified formula and modified low-protein foods). When women with PKU do not adhere to their diet before and during pregnancy, infants born to them have a 93% risk for MR and a 72% risk for microcephaly. These risks result from the toxic effects of high maternal blood phe levels during pregnancy, not because the infant has PKU. The restricted diet, which should be maintained for life, often is discontinued during adolescence. This report describes the pregnancies of three women with PKU and underscores the importance of overcoming the barriers to maintaining the recommended dietary control of blood phe levels before and during pregnancy. For maternal PKU-associated MR to be prevented, studies are needed to determine effective approaches to overcoming barriers to dietary control. PMID- 11898926 TI - Measles--United States, 2000. AB - In 2000, a provisional total of 86 confirmed measles cases were reported to CDC by state and local health departments, representing a record low and a 14% decrease from the 100 cases reported in each of the previous 2 years. This report describes the epidemiology of measles in the United States during 2000 and documents the continued absence of endemic measles and the continued risk for internationally imported measles cases that might result in indigenous transmission. PMID- 11898927 TI - State-specific mortality from sudden cardiac death--United States, 1999. AB - Each year in the United States, 400,000-460,000 persons die of unexpected sudden cardiac death (SCD) in an emergency department (ED) or before reaching a hospital (1). Based on the latest U.S. mortality data, this report summarizes and analyzes 1999 national and state-specific SCD data. Reducing the proportion of out-of hospital* SCDs would decrease the overall incidence of premature death in the United States. Heart attacks are the major cause of SCD; approximately 70% of SCDs are caused by coronary heart disease. National efforts are needed to increase public awareness of heart attack symptoms and signs and to reduce delay time to treatment. PMID- 11898928 TI - The better-than-nothing idea: debating the use of placebo controls. PMID- 11898929 TI - Community health programs in Canada. PMID- 11898930 TI - Ethics and industry-sponsored research. PMID- 11898931 TI - Ethics and industry-sponsored research. PMID- 11898932 TI - Ethics and industry-sponsored research. PMID- 11898933 TI - Ethics and industry-sponsored research. PMID- 11898934 TI - Ethics and industry-sponsored research. PMID- 11898935 TI - Ethics and industry-sponsored research. PMID- 11898936 TI - PSA screening and prostate cancer mortality. AB - BACKGROUND: Physicians have speculated that prostate-specific antigen (PSA) screening may be responsible for the reduction in prostate cancer mortality observed in the late 1 990s. In order to test this hypothesis, we assessed the relation between the change in prostate cancer incidence in the early 1990s, attributed largely to PSA screening, and the subsequent change in prostate cancer mortality. METHODS: We divided the adult male population of Quebec aged 50 years and more into 15 birth cohorts. For each birth cohort, we computed the change in prostate cancer incidence between 1989 and 1993 and the change in prostate cancer mortality between 1995 and 1999. We then assessed the correlation between the changes in prostate cancer incidence and the subsequent changes in prostate cancer mortality by weighted linear regression. We also split up the study population into 15 regional populations and repeated the analysis described above. RESULTS: We found that even though most birth cohorts showed an increase in prostate cancer incidence and a subsequent decrease in mortality, the sizes of these changes were not inversely correlated (Pearson's r = 0.33, 1-sided p = 0.89). Similarly, in the regional population study, we found that a greater increase in prostate cancer incidence did not indicate a greater decline in mortality (Pearson's r= 0.13, 1-sided p = 0.68). INTERPRETATION: These results suggest that for our study population PSA screening was not associated with, and therefore cannot explain, the decline in prostate cancer mortality. PMID- 11898938 TI - Cervical intraepithelial neoplasia in women presenting with external genital warts. PMID- 11898937 TI - Effect of legislation on the use of bicycle helmets. AB - BACKGROUND: About 50 Canadian children and adolescents die each year from bicycle related injuries, and 75% of all bicycle-related deaths are due to head injuries. Although the use of helmets can reduce the risk of head injury by 85%, the rate of voluntary helmet use continues to be low in many North American jurisdictions. We measured compliance before, during and after 1997, when legislation making the use of helmets mandatory for cyclists was enacted in Nova Scotia. METHODS: In the summers and autumns of 1995 through 1999, trained observers who had a direct view of oncoming bicycle traffic recorded helmet use, sex and age group of cyclists in Halifax on arterial, residential and recreational roads. Sampling was done during peak traffic times of sunny days. We abstracted data from the Canadian Hospitals Injury Reporting and Prevention Program database on bicycle-related injuries treated during the same period at the Emergency Department of the IWK Health Centre, Halifax. RESULTS: The rate of helmet use rose dramatically after legislation was enacted, from 36% in 1995 and 38% in 1996, to 75% in 1997, 86% in 1998 and 84% in 1999. The proportion of injured cyclists with head injuries in 1998/99 was half that in 1995/96 (7/443 [1.6%] v. 15/416 [3.6%]) (p = 0.06). Police carried out regular education and enforcement. There were no helmet promoting mass media education campaigns after 1997. INTERPRETATION: Rates of helmet use rose rapidly following the introduction of legislation mandating the use of helmets while bicycling. The increased rates were sustained for 2 years afterward, with regular education and enforcement by police. PMID- 11898939 TI - Does PSA screening reduce prostate cancer mortality? PMID- 11898940 TI - Hats off (or not?) to helmet legislation. PMID- 11898941 TI - Placebo trials and tribulations. PMID- 11898942 TI - Clinical nutrition: 4. Omega-3 fatty acids in cardiovascular care. PMID- 11898943 TI - Advances in the pharmacotherapy of Alzheimer's disease. PMID- 11898944 TI - Osteoarticular tuberculosis: a case report and discussion. PMID- 11898945 TI - Varicella control and vaccine coverage: issues and challenges. PMID- 11898946 TI - Generalized peritonitis. PMID- 11898947 TI - Ephedra/ephedrine: cardiovascular and CNS effects. PMID- 11898948 TI - Anne McLellan: new direction from a new minister? PMID- 11898949 TI - "No statute remains frozen in time forever": Romanow. PMID- 11898950 TI - Canada to reconsider ethics of placebo-controlled trials. PMID- 11898951 TI - In Iran, gender segregation becoming a fact of medical life. PMID- 11898952 TI - Improvement in current approaches to lipid lowering. PMID- 11898953 TI - End-of-life care. PMID- 11898954 TI - Perinatal HIV testing. PMID- 11898955 TI - Physicians should not be referred to as 'providers'. PMID- 11898956 TI - Osteoarthritis: diagnosis and therapeutic considerations. AB - Osteoarthritis is a common rheumatologic disorder. It is estimated that 40 million Americans and 70 to 90 percent of persons older than 75 years are affected by osteoarthritis. Although symptoms of osteoarthritis occur earlier in women, the prevalence among men and women is equal. In addition to age, risk factors include joint injury, obesity, and mechanical stress. The diagnosis is largely clinical because radiographic findings do not always correlate with symptoms. Knowledge of the etiology and pathogenesis of the disease process aids in prevention and management. Acetaminophen and nonsteroidal anti-inflammatory medications remain first-line drugs. Agents such as cyclooxygenase-2 inhibitors and sodium hyaluronate joint injections offer new treatment alternatives. Complementary medication use has also increased. Therapeutic goals include minimizing symptoms and improving function. PMID- 11898958 TI - Information from your family doctor. Hereditary hemochromatosis. PMID- 11898957 TI - Recognition and management of hereditary hemochromatosis. AB - Hereditary hemochromatosis is the most common inherited single-gene disorder in people of northern European descent. It is characterized by increased intestinal absorption of iron, with deposition of the iron in multiple organs. Previously, the classic description was combined diabetes mellitus, cutaneous hyperpigmentation and cirrhosis. Increasingly, however, hereditary hemochromatosis is being diagnosed at an earlier, less symptomatic stage. The diagnosis is based on a combination of clinical, laboratory and pathologic findings, including elevated serum transferrin saturation. Life expectancy is usually normal if phlebotomy is initiated before the development of cirrhosis or diabetes mellitus. Hereditary hemochromatosis is associated with mutations in the HFE gene. Between 60 and 93 percent of patients with the disorder are homozygous for a mutation designated C282Y. The HFE gene test is useful in confirming the diagnosis of hereditary hemochromatosis, screening adult family members of patients with HFE mutations and resolving ambiguities concerning iron overload. PMID- 11898959 TI - Cholesterol treatment guidelines update. AB - Hypercholesterolemia is one of the major contributors to atherosclerosis and coronary heart disease in our society. The National Cholesterol Education Program of the National Institutes of Health has created a set of guidelines that standardize the clinical assessment and management of hypercholesterolemia for practicing physicians and other professionals in the medical community. In May 2001, the National Cholesterol Education Program released its third set of guidelines, reflecting changes in cholesterol management since their previous report in 1993. In addition to modifying current strategies of risk assessment, the new guidelines stress the importance of an aggressive therapeutic approach in the management of hypercholesterolemia. The major risk factors that modify low density lipoprotein goals include age, smoking status, hypertension, high-density lipoprotein levels, and family history. The concept of "CHD equivalent" is introduced-conditions requiring the same vigilance used in patients with coronary heart disease. Patients with diabetes and those with a 10-year cardiac event risk of 20 percent or greater are considered CHD equivalents. Once low-density lipoprotein cholesterol is at an accepted level, physicians are advised to address the metabolic syndrome and hypertriglyceridemia. PMID- 11898960 TI - Identifying and managing preparatory grief and depression at the end of life. AB - Grief and depression present similarly in patients who are dying. Conventional symptoms (e.g., frequent crying, weight loss, thoughts of death) used to assess for depression in these patients may be imprecise because these symptoms are also present in preparatory grief and as a part of the normal dying process. Preparatory grief is experienced by virtually all patients who are dying and can be facilitated with psychosocial support and counseling. Ongoing pharmacotherapy is generally not beneficial and may even be harmful to patients who are grieving. Evidence of disturbed self-esteem, hopelessness, an active desire to die and ruminative thoughts about death and suicide are indicative of depression in patients who are dying. Physicians should have a low threshold for treating depression in patients nearing the end of life because depression is associated with tremendous suffering and poor quality of life. PMID- 11898961 TI - Information from your family doctor. Dying and preparatory grief. PMID- 11898962 TI - Lipoma excision. AB - Lipomas are adipose tumors that are often located in the subcutaneous tissues of the head, neck, shoulders, and back. Lipomas have been identified in all age groups but usually first appear between 40 and 60 years of age. These slow growing, nearly always benign, tumors usually present as nonpainful, round, mobile masses with a characteristic soft, doughy feel. Rarely, lipomas can be associated with syndromes such as hereditary multiple lipomatosis, adiposis dolorosa, Gardner's syndrome, and Madelung's disease. There are also variants such as angiolipomas, neomorphic lipomas, spindle cell lipomas, and adenolipomas. Most lipomas are best left alone, but rapidly growing or painful lipomas can be treated with a variety of procedures ranging from steroid injections to excision of the tumor. Lipomas must be distinguished from liposarcoma, which can have a similar appearance. PMID- 11898963 TI - Information from your family doctor. What are lipomas? PMID- 11898964 TI - Rhabdomyolysis. AB - Rhabdomyolysis is a potentially life-threatening syndrome resulting from the breakdown of skeletal muscle fibers with leakage of muscle contents into the circulation. The most common causes are crush injury, overexertion, alcohol abuse and certain medicines and toxic substances. Several inherited genetic disorders, such as McArdle's disease and Duchenne's muscular dystrophy, are predisposing factors for the syndrome. Clinical features are often nonspecific, and tea colored urine is usually the first clue to the presence of rhabdomyolysis. Screening may be performed with a urine dipstick in combination with urine microscopy. A positive urine myoglobin test provides supportive evidence. Multiple complications can occur and are classified as early or late. Early complications include severe hyperkalemia that causes cardiac arrhythmia and arrest. The most serious late complication is acute renal failure, which occurs in approximately 15 percent of patients with the syndrome. Early recognition of rhabdomyolysis and prompt management of complications are crucial to a successful outcome. PMID- 11898965 TI - Maternal serum triple analyte screening in pregnancy. AB - According to the American College of Obstetricians and Gynecologists, it has become standard in prenatal care to offer screening tests for neural tube defects and genetic abnormalities. There have been some changes in the recommended method of prenatal screening over the past few years, and research to improve detection rates with better combinations of maternal serum analytes is ongoing. The issues facing physicians are the sensitivity and specificity of multiple serum analyte combinations. The current maternal serum analytes in use in most areas are alpha fetoprotein (AFP), human chorionic gonadotropin (hCG) and unconjugated estriol. Measurement of AFP alone can detect the vast majority of neural tube defects and a small portion of trisomy 21-affected pregnancies in patients of all ages. Adding hCG and unconjugated estriol to this screen increases the rate of detection of trisomies 21 and 18. Counseling patients about the risks and benefits of such screening is important to provide a balanced discussion of screening issues. PMID- 11898966 TI - Information from your family doctor. Triple screening in pregnancy--what it is and what to expect. PMID- 11898967 TI - Low back pain. PMID- 11898968 TI - AAP updates statement for transfer of drugs and other chemicals into breast milk. American Academy of Pediatrics. PMID- 11898969 TI - New antiretroviral for HIV infection. PMID- 11898970 TI - A study of parents of violent children. AB - This study based on in-depth interviews of 25 parents of violent children and a control group of 25 parents of nonviolent children concerned the parents' personalities. Parents were between 22 and 48 years of age and were from middle and lower middle socioeconomic backgrounds. Differences in classification by the nonblinded interviewers of parents into the two groups on six behavior characteristics were significant on chi2 tests. Some recommendations are made for further research. PMID- 11898971 TI - Note on reliability of the Carlson Psychological Survey. AB - The Carlson Psychological Survey is designed to assess and classify criminal offenders but its reliability has not been estimated for use with female adolescents. The survey was administered to 36 female adjudicated delinquents between the ages of 14 and 16 years and readministered after a 2-wk. interval to estimate test-retest reliability. Support for its stability over 2 wk. with this specific adolescent population was obtained. PMID- 11898972 TI - High risk behaviors in a sample of Mexican-American college students. AB - This study explored different types of high risk behaviors of Mexican-American college students attending a small university in south Texas. High risk behaviors for contracting HIV/AIDS examined in this study included unprotected sex, drug use, and alcohol abuse. In 1995 in the United States, HIV/AIDS is the leading cause of death in people between the ages of 25 and 44. Because use of alcohol and certain recreational drugs lowers inhibitions, their use could increase the possibility of having unprotected and unplanned sex with multiple partners. Thus, it was expected that Mexican-American college students who use drugs and alcohol would be more likely to engage in unprotected sex. Data were from 105 men and 211 women between the ages of 18 and 30 years. Drug use and alcohol abuse were significantly associated with high risk sexual behavior. Individuals in monogamous relationships were more likely to not use condoms than those involved in casual relationships. Self-reported religiosity was not correlated with high risk behaviors, although there were implications that stronger religious affiliation did alter sexual beliefs and practices. Lastly, parental communication was not significantly associated with high risk behaviors, but family unity did seem related to some risky sexual practices. PMID- 11898973 TI - Specificity of the MMPI-2 Fake Bad Scale as a marker for personal injury malingering. AB - Psychologists who evaluate patients in medicolegal contexts should utilize objective assessment data with empirically established sensitivity and specificity for identifying negative response bias. The purpose of this study was to investigate the specificity of the Fake Bad Scale for identifying negative response bias in personal injury claimants. The cutoff scores proposed by Lees Haley and colleagues were applied a federal prison, medical outpatients, and patients from to inmate volunteers from substance abuse unit. Half of the inmates were given instructions to malinger psychopathology to affect the adjudication process, and the remaining inmates and all of the hospital patients were given standard instructions. The original cutoff scores correctly identified the majority of inmates instructed to malinger psychopathology, but these scores resulted in unacceptably high rates of false positive classifications. The revised cutoff scores resulted in fewer false positives, i.e., 8%-24%. PMID- 11898974 TI - A quantitative method for studying the testing supervision process. AB - A quantitative method by Holloway for studying the supervision process in counseling training was adapted for use in psychological testing supervision. Ratings of 80 sessions indicated that most of the 25 potential interactions between the supervisor and trainee occurred in most of the sessions. The rate of occurrence was the lowest for the Emotional Awareness task (21%-24%). The author offers a speculation about how this result may be understood as involving the structured nature of testing. Results suggested that this model could be used as a tool for both practical and theoretical empirical evaluation of the supervision process during training in psychological testing. PMID- 11898975 TI - Influence of prior involvement on employees' ratings. AB - Surveys show that the majority of firms provide merit pay to outstanding employees. Despite the widespread use of merit pay programs, there is no consensus as to their effectiveness. While some plans have been successful in motivating employees to achieve higher performance, others have caused employees' dissatisfaction and discouragement. One previously unexplored area in the implementation of merit pay systems is the effect of the decision-maker's prior involvement with the employee on the decision to grant merit. The purpose of this paper was to examine whether hiring an employee leads to larger merit allocations despite evidence that the standards for merit were not met. 101 accounting students participated to assess the influence of hiring on subsequent merit allocations. Analysis indicated that prior involvement does bias ensuing merit decisions. PMID- 11898976 TI - Accessing directorships: comparison of views of Canadian and Australian women directors. AB - Compared are views of Canadian and Australian women directors concerning the difficulties women face in accessing the most privileged level of management directorships of companies. The Canadian data are from a study of 278 women directors of corporate boards in Canada while the Australian results are from a study of 47 women directors of publicly listed companies in Australia. Despite the different time periods and geographical locations in which the studies were carried out, the profiles and responses of the two groups are quite similar. Both groups believe the current mix of directors is not adequate and that barriers still exist in nominating women to boards. PMID- 11898977 TI - Teachers' reports of the problem behavior of children in their classrooms. AB - Research examining teachers' judgments of children's behavior has typically used archival data, staged videos, or written vignettes describing children's behavior. The main advantage of using staged videos and written vignettes has been that those methodologies have led to well-controlled studies. The main disadvantage is that little is known about teachers' perceptions of the problems of children in their own classrooms. In the current study. 111 first-, second-, and third-grade teachers described children in their classrooms whose behavior concerned them. Teachers identified significantly more children with externalizing problems than internalizing problems and significantly more boys than girls as having problems that concerned them. However, when teachers identified children as having internalizing problems, they were just as likely to judge them as needing referral as children with externalizing problems. Similarly, when teachers judged children to have problems that concerned them, they were just as likely to judge girls as needing referral as boys. PMID- 11898978 TI - Do homosexual teachers account for about half of news stories of molestations of pupils? A Boston Globe replication. AB - Homosexual interaction was involved in 11 (48%) of 23 and 10 (45%) of 22, that is, about half of two nationwide databases of newspaper stories about teachers sexual involvement with pupils reported by Cameron and Cameron in 1998. Whether this relationship holds at a local level was examined by searching all indexed 'sex crimes' in the Boston Globe from 1991 through 1998 for local stories about sex between pupil and teacher. Of the 21 teachers in 20 stories, 11 (52%) interacted homosexually with pupils. Thus it appears that nationally and locally, as reported in newspapers, about half of the molestations by teachers are homosexual. PMID- 11898979 TI - Validity of projective drawing indices of male homosexuality. AB - We investigated the hypothesis that certain signs in the Draw-A-Person projective technique reflect male homosexuality. Human figure drawings from gay and 88 heterosexual men, with no clinical psychological symptoms, were submitted to trained raters who were blind to the purpose of this research. The raters independently judged whether 21 signs, previously referenced in the literature as suggestive of male homosexuality, were present in the figure drawings. Only two signs, hair and hips, differentiated between groups. Results were interpreted in the light of changing social attitudes towards homosexuals and methodological problems of prior studies. PMID- 11898980 TI - Effect of forced laughter on mood. AB - This study examined the effect of a brief period of forced laughter on the mood of adults. Participants (N=17) rated their mood before and after 1 min. of forced laughter. Although the participants generally rated their mood as positive prior to the intervention, after forced laughter more participants rated positive affect significantly higher. PMID- 11898981 TI - Sex differences in symptoms of depression among American children and adolescents. AB - The Arabic Children's Depression Inventory in its English version was administered to a sample of 535 U.S. school students (11 to 18 years old). By sex, differences on total score and on 19 of 27 items (70.4%) were not statistically significant. Nevertheless, differences were significant for 8 (29.6%) items on which girls had higher mean scores. Half of the items were positive indicators of depression (I am sad, I feel lonely, I feel miserable, and I hate myself), while the other half were negative (I feel happy, Life is rosy, A lot of people like me, and I am optimistic). PMID- 11898983 TI - Customer service as a function of shopper's attire. AB - A field experiment explored whether a female shopper's appearance would influence the customer service she received. Specifically, a female confederate dressed in formal work clothes (skirt and blouse) or informal gym clothes (tights and t shirt) entered a series of randomly selected women's clothing stores in a large mall and proceeded to "shop." The amount of time that passed before an employee approached and acknowledged the confederate served as the dependent variable. As hypothesized, she was acknowledged significantly sooner when formally attired than when informally dressed. Thus, clothing, like other aspects of appearance, influences how people are evaluated and treated by others. PMID- 11898982 TI - Toward the content validity of premenstrual dysphoric disorder: do anger and irritability more than depressed mood represent treatment-seekers' experiences? AB - The content validity of Premenstrual Dysphoric Disorder in the Diagnostic and Statistical Manual of Mental Disorders, fourth edition, text revision (DSM-IV-TR) has been questioned in the literature. We tested whether mood-related symptoms reported by 26 women seeking treatment for premenstrual disorders were among the proposed criteria. These women were asked to list their premenstrual symptoms and rate the severities of listed symptoms daily for two menstrual cycles before treatment. They completed semistructured interviews to differentiate symptoms of Premenstrual Dysphoric Disorder from those of other psychiatric disorders in women who had other disorders. All participants reported functional interference due to the symptoms. 19 symptoms of or similar to those of Premenstrual Dysphoric Disorder were among the 22 most frequent premenstrual symptoms experienced. Premenstrual depressed mood was less frequent than premenstrual irritability or anger when other psychiatric disorders such as major depression were taken into account. Results suggest that the DSM-IV-TR criteria have generally good content validity but may need revision to represent treatment-seekers experiences more accurately. PMID- 11898984 TI - Alexithymia and sociocultural factors in a Japanese sample: a study with the Rorschach. AB - We reexamined the Japanese version of the 20-item Toronto Alexithymia Scale, a self-report sc ale for measuring alexithymic characteristics, by comparing the scores on three factors and the total scores with variables of the Rorschach in a sample of 40 (originally 48) Japanese college students. Based on prior studies, our aims were to further validate the 20-item Toronto Alexithymia Scale by comparing its scores with those on a projective technique. We also investigated whether sociocultural factors, such as repression of hostility. are associated with scores on the 20-item Toronto Alexithymia Scale (especially Factor 3). None of the seven Rorschach Alexithymia Variables were significantly related to the factors and total scores of the 20-item Toronto Alexithymia Scale. However, scores for Factor 1 (difficulty identifying feelings) of the 20-item Toronto Alexithymia Scale correlated positively with scores on Sum C' (reserved responses to emotional stimuli) and Adj es (stimulus demand), suggesting that individuals who score high for Factor 1 experience gloomy, depressive feelings but in constricted ways. Scores for Factor 3 (externally oriented thinking) correlated positively with the D scores (stress tolerance) and negatively with m (situational stress) responses, suggesting that Factor 3 may reflect a psychological defense that enhances stress tolerance. There was a nonsignificant negative correlation between Factor 1 and Factor 3 scores, but, unlike our hypothesis. Factor 3 was neither related to AG (aggression) nor S (space responses reflecting oppositional tendency), indices of aggression or hostility in the Rorschach Comprehensive System. It may be that the 20-item Toronto Alexithymia Scale and the Rorschach measure quite different aspects of personality, but further research is necessary. PMID- 11898985 TI - The father-daughter relationship rating scale. AB - A 9-item scale designed to measure perceived relationships of girls and their fathers is internally consistent (alpha=.89) and showed clear factor structure. PMID- 11898986 TI - Perception in consequences of free riding. AB - This study examined the relationships between perceptions of group members' free riding and group outcomes using Mulvey and Klein's 1998 perceived free riding scale. In a laboratory study, three free riding conditions were created (no free riding, free riding, free riding with justification) in which 97 college students performed two short number-finding tasks as members of temporary ad hoc three person groups. 55% of the students were male and the average age was 22.9 yr. (SD= 3.0). Participants' perceptions of free riding were negatively related to commitment to the assigned group goal, task performance, and goals for group performance and individual performance. In the condition wherein free riding was justified by low ability, participants set lower goals for their future task performance than did those in the other two conditions. PMID- 11898987 TI - Immediate Feedback Assessment Technique: multiple-choice test that "behaves" like an essay examination. AB - A multiple-choice testing system that provides immediate affirming or corrective feedback and permits allocation of partial credit for proximate knowledge is suggested as an alternative to essay examinations. PMID- 11898988 TI - Correlations for scores on the 180-item version of the MMPI-2 and the Neuroticism scale of the NEO-Personality Inventory. AB - 64 normal adult controls (53 men, 11 women; M age 45.2 yr.) in a study of an estuary-associated syndrome were administered the MMPI-2-180 and the Neuroticism scale from the NEO-Personality Inventory. Pearson product-moment correlations between MMPI-2 scales and the Neuroticism scale were similar to those reviously reported using the full-length MMPI. Correlations between MMPI-2 scales, D, Pt, Sc, and Si, and NEO-PI Neuroticism (range .44 to .52) suggest that many psychiatric conditions are associated with psychological distress of the type individuals high in trait neuroticism are prone to experience. PMID- 11898989 TI - Attachment style and self-reported aggression. AB - The present study examined the relationship between attachment style and self reported aggression in a sample of young women (N = 139). Analysis showed that 27.3% of the subjects classified themselves as insecurely, i.e., avoidantly or ambivalently, attached by means of the Adult Attachment Questionnaire of Hazan and Shaver. In addition, the insecurely attached subjects had elevated anger and hostility compared to securely attached subjects. PMID- 11898990 TI - Apparent higher brain weight in suicide victims: possible reasons. AB - In 2000 we tested previously reported findings by Salib and Tadros that brain weight of fatal self-harm victims is higher than of those who died of natural causes. Our results were based on data from 15 suicides and 15 deaths of other causes. Data included matching variables of age, sex, time between death and postmortem examination, and temperature of the surrounding environment. The exploratory variables were brain weight and method of death. No significant difference was found between the brain weights of suicides and others. On the other hand, some differences were obtained for different suicide methods, which also differed in the temperature of the environment, this being lower for the group of suicides that occurred outdoors (around or below 0 degrees C). Once we excluded all the outdoor cases and controls, a significantly higher brain weight was obtained for suicide cases. These and previous results are intriguing and require explanation. Respirator brain syndrome as described by Moseley, Molinari, and Walker in 1976 may provide only a partial explanation. Another possible suggestion is that higher brain weight in suicide victims may be related to previously demonstrated increased amygdala blood flow and subsequent amygdala enlargement due to the increased processing of emotional information. PMID- 11898991 TI - Locus of control of children experiencing separation and divorce in their families in Iran. AB - A comparison of scores on locus of control by three groups of children in intact families (n=676), parent loss through death (n=30), and parent loss through divorce (n = 20) showed that children from divorced families scored significantly more external than children from intact families. The results are considered in relation to previous studies and some educational implications noted. PMID- 11898992 TI - A preliminary investigation of multicultural perspective and life satisfaction. AB - The present study represents a preliminary investigation of Ramirez' contention that developing a multicultural identity would lead to better mental health and more life satisfaction. Using a tentative index of a multicultural perspective analysis indicated that, although a multicultural perspective is associated with more life satisfaction, it accounts for only 8% of the variance. Further, one of the measures used by Ramirez to measure an aspect of a multicultural perspective was not correlated with life satisfaction, and another correlated negatively. This latter measure accounts for only 6% of the variance in life satisfaction. Combined, these findings provide limited support for Ramirez perspective by explaining 14% of the overall variance in life satisfaction. PMID- 11898993 TI - Relationship between stages of smoking acquisition and environmental factors among junior high school students. AB - To study the relationship of stages of smoking acquisition and environmental factors, 757 Japanese junior high school students (390 boys and 367 girls) anonymously answered a questionnaire regarding the four stages of smoking acquisition (precontemplation, contemplation, preparation, and action stages) and environmental factors of smoking behavior. Analysis showed that the influence of advertising and students' interpersonal situations on smoking behavior increased from the precontemplation stage to the action stage. PMID- 11898994 TI - Mental health among Afghan refugees settled in Shiraz, Iran. AB - This study was designed to investigate the mental health of Afghan refugees settled in Shiraz, the capital of a southern province of Iran. They were mostly refugees from Afghanistan by reason of internal war during the last two decades. A version of the General Health Questionnaire (GHQ-28) in Persian was administered on a group of randomly selected Afghan refugees (n=81) from a pool of Afghan residents in the Shiraz district. 34.5% of the subjects scored high enough to be considered as having psychiatric problems. There was a significant positive correlation between refugees' years of age and GHQ-28 factor scores, i.e., Physical Health and Social Functioning. The mental health of the subjects was not related to education or marital status. The years of settling in Iran were not significantly correlated with any GHQ-28 indices. The overall findings suggest that the rate of psychiatric problems in the refugees is higher than in the native population. PMID- 11898995 TI - An evaluation of touch on a large request: a field setting. AB - The effect of touch on compliance to a request has traditionally been tested with small solicitation (answer to a small questionnaire, give a dime to a confederate ....). In our experiment a larger request was evaluated. Passersby, 53 men and 67 women, were asked by two confederates to look after a large and very excited dog for 10 minutes because each wanted to go into a pharmacy where animals were prohibited. In half of the cases, subjects were touched during the request. Analysis showed that, when touched, 55% of the subjects agreed with the request whereas 35% only in the no-touch control condition agreed. This finding indicates that touch was positively associated with the subjects' compliance (p<.03). PMID- 11898996 TI - Sex, perception of immediate stress, and response to coping resources inventory, emotional domain. AB - The purpose was to assess differences in mean scores of 23 men and 28 women who rated immediate stress low and 11 men and 34 women who rated immediate stress high. Analysis of variance of mean differences was significant for sex but not for immediate stress or their interaction on the Coping Resources Inventory, Emotional Domain. PMID- 11898997 TI - An approach to the hierarchical model of motivation in a classroom: a reply to Rousseau and Vallerand. AB - Rousseau and Vallerand's comments on our recent article led to the present study of correlations between students' perception of teachers' attitudes, intrinsic extrinsic motivation, and learning strategy with structural equation modeling. The analysis shows students' perception of teachers' attitudes influences the intrinsic-extrinsic motivation, which as mediator in turn affects students' learning strategies, and the paths were divided into two streams, one with more intrinsic attributes and the other with more extrinsic ones. PMID- 11898998 TI - Adult attachment interview, thematic analysis, and communicative style in families with substance use disorder. AB - The paper examined the Adult Attachment Interview with special reference to thematic and semantic analysis in line with the discourse study (van Dijk, 1997). The hypothesis was that correspondence between the communicative organization of speech and the mental representations of the attachment experiences would be substantial. Eight Adult Attachment Interview transcripts of fathers with a heroin addicted young son were analyzed at two levels, (a) thematic analysis to individuate the topics of their talk applying the structural and semantic study of discourse and (b) enunciative analysis of speech to define their linguistic patterns utilizing a set of linguistic micro- and macro-units. Results showed nine main topics in the Adult Attachment Interview, each of which was characterized by a distinctive linguistic profile. In this perspective this device seems to be effective not only for discriminating attachment styles between subjects but also to identify differences within subjects belonging to the same attachment pattern. PMID- 11898999 TI - Prediction of graduate major from cognitive and self-report test scores obtained during the high school years. AB - Using information from Project TALENT's 11-yr. follow-up (Wise, et al. 1979), 8 graduate major groups were formed for males, 6 for females. Three of the female groups were supplemented by undergraduate majors to increase sample size. Descriptive canonical discriminant analyses were conducted in each sex, high school grade, and separate sets of cognitive and self-report predictors that were administered in high school. Finally, analyses of the combined predictors were conducted in both sexes for the 9th grade only. Two to four canonical discriminants were retained for interpretation on two criteria: lambda probabilities and size of eigenvalues. Discrimination among groups is more effective in males than in females and in later than in earlier grades. Self report tests discriminate more effectively among graduate major groups than cognitive ones in the cognitively restricted range of talent. Gains in differentiating among groups from adding cognitive to self-report predictors in the 9th grade are relatively modest. The two sets of predictors define overlapping if not identical dimensions. The first two cognitive functions are important supplements to a measure of general intelligence. For both sexes, a combination of mechanical, spatial, and mathematical tests distinguish science majors from those in the humanities and social sciences who appear on a verbal function. PMID- 11899000 TI - Collectivism and rates of personal violence (suicide and homicide). AB - Homicide rates, but not suicide rates, were associated with an index of collectivism for the states of America in 1992. PMID- 11899001 TI - Effects of generalized self-efficacy and negative social comparison feedback on specific self-efficacy and performance. AB - The effects of social comparison feedback on specific self-efficacy and performance of high generalized self-efficacy participants and low generalized self-efficacy participants were examined with the help of 20 participants with high generalized self-efficacy and 20 participants with low generalized self efficacy. Half of the participants in each generalized self-efficacy group received negative social comparison feedback after each of four trials of an experimental task while the other half received no feedback. Two kinds of specific self-efficacy-performance-based and normative-based--were measured once before the task and four times after the trials of the task. After the task, the High generalized self-efficacy/Feedback group rated performance-based specific self-efficacy higher and performed better than the Low generalized self efficacy/Feedback group. No significant difference was observed between the High generalized self-efficacy/No feedback group and Low generalized self-efficacy/No feedback group. There were no significant effects with regard to normative-based specific self-efficacy. PMID- 11899002 TI - Burnout syndrome in the helping professions. AB - Burnout can be defined as a long-term reaction to occupational stress which involves, particularly, the helping professions. The main aim of this study was the assessment of burnout in a sample of professional and voluntary health care workers and comparison of the two samples on scores from the Maslach Burnout Inventory. Analysis suggests a significant difference in mean scores for Emotional Exhaustion of volunteers vs professional workers. Some evidence has supported the hypothesis of a fourth dimension, called Behavioral Exhaustion, in the burnout syndrome. PMID- 11899003 TI - Revising the Competitiveness Index using factor analysis. AB - The Competitiveness Index is a 20-item true-false measure designed to assess the desire to win in interpersonal situations. To develop a more psychometrically sound form of the scale, 213 undergraduates were administered the original form and a modified version containing a 5-point Likert-type scale. An initial principal component analysis using a varimax rotation of the modified version yielded a four-factor solution accounting for 54.5% of the explained variance. Based on a subsequent reliability analysis. six items were dropped from the modified scale. A second analysis produced a two-factor solution accounting for 54.1% of the explained variance. Both factors (Enjoyment of Competition and Contentiousness) formed reliable subscales. The 14-item Revised Competitiveness Index had high internal consistency and was positively correlated with the original Competitiveness Index, the competitiveness subscales of the Work and Family Orientation Questionnaire, the Sports Orientation Questionnaire, and the Nach Naff measure of Need for Achievement. PMID- 11899004 TI - Psychometric properties of the statistics anxiety rating scale. AB - The Statistics Anxiety Rating Scale has 51 items, each scored on a 5-point rating scale to measure statistics anxiety with six subscales, Worth of Statistics, Interpretation Anxiety, Test and Class Anxiety, Computational Self-concept, Fear of Asking for Help, and Fear of Statistics Teachers. Psychometric properties included analyses of construct and concurrent validities an internal consistency and test-retest reliability. 221 college students (74% women; M age=28 yr.) in elementary statistics courses at several southwestern state universities participated. The findings are consistent with previous reports and indicate adequate concurrent validity, internal consistency, and split-half reliability, but for construct validity confirmatory factor analysis yielded marginal support. PMID- 11899005 TI - Presidential debates and change in students' attitude. AB - The effects of debates on influencing potential voters' attitudes were assessed in a group of 45 undergraduates who watched the third presidential debate of election year 2000 between candidates Bush and Gore. A repeated measures t test indicated a significant change in immediate ratings of attitude from pretest to posttest, with Gore being rated higher at posttest. PMID- 11899006 TI - The interactive effects of low self-control and commitment to school on substance abuse among college students. AB - This study examined the combined influence of two predicting factors-low self control and commitment to schooling-that research has shown have independent effects on substance abuse. In a sample of 598 college students, this study tested the interactive effects of these factors while controlling for other established predictors of binge drinking and drug use. Analysis showed that participants who had both low self-control and low schooling commitment had significantly higher scores on substance abuse than would be expected from the independent influences of the component factors, which suggests that the combined effects of these predictors on substance abuse have a greater influence than their direct influences. PMID- 11899007 TI - School characteristics among children of alcoholic parents. AB - Studies show that children of alcoholics constitute a population at-risk commonly for poor performance, skipping school days, and school drop out. The focus of the present study was to examine a variety of direct outcome variables measuring academic performance among a sample of 226 children, 108 of them from parents who misused alcohol in Cadiz. Parents were outpatients of a Health Service and received treatment for the drinking problem; 118 students were children of nonalcoholic parents attending the same schools as the children of alcoholic parents. Both groups were compared on age, sex, school grade, and social environment. The study identified five variables on which performance by children of alcoholic parents was poorer: intelligence, repeating a grade, low academic performance, skipping school days, and dropping out of school. PMID- 11899008 TI - Social facilitation in young toddlers. AB - As a test of Zajonc's social facilitation hypothesis, the behavior of 18 young toddlers (16-24 mo.) was observed. Their performance on a simple task was incongruent with his hypothesis, that is, these young toddlers may not evaluate their outcomes in relation to a social standard. PMID- 11899009 TI - Evidence for an association between gender-role identity and a measure of executive function. AB - Two studies assessed the relation between gender role and executive function. In Study One (N = 234) analyses indicated that among college students executive function, assessed by the Coolidge and Griego scale, is related to masculine gender-role classification, measured by the Bem Sex-role Inventory. This relationship remained significant when biological sex was controlled. Further, factor analysis of the Bem Sex-role Inventory identified six components, three related to executive function. Two of these scales were associated with masculine characteristics, and the third was associated with the denial of several feminine items. Study Two (N = 53) further assessed the relationship among undergraduates through additional measures of executive functions and mood, in addition to the Bem Sex-role Inventory. In this study, executive functioning, as measured by the Coolidge and Griego scale, was again generally related to masculinity. Psychological well-being was not related to gender identity or executive functioning. PMID- 11899010 TI - Client perspectives and open-ended questions: a reply to Dean, et al. (2000). AB - Utilizing an open-ended question may allow subjects to respond in their own words but does not necessarily avoid the problem of narrowing the focus to a specific predetermined topic. PMID- 11899011 TI - A preliminary study of achievement, attitudes toward success in mathematics, and mathematics anxiety with technology-based instruction in brief calculus. AB - This study was designed to compare achievement, attitudes toward success in mathematics, and mathematics anxiety of college students taught brief calculus using a graphic calculator, with the achievement and attitudes and anxiety of students taught using the computer algebra system Maple, using a technology based text book. 50 men and 50 women, students in three classes at a large public university in the southwestern United States, participated. Students' achievement in brief calculus was measured by performance on a teacher-made achievement test given at the end of the study. Analysis of variance showed no significant difference in achievement between the groups. To measure change in attitudes and anxiety, responses to paper-and-pencil inventories indicated significant differences in favor of students using the computer. PMID- 11899012 TI - Voluminous sucrose consumption in female rats: increased "nippiness" during periods of sucrose removal and possible oestrus periodicity. AB - In a two-bottle paradigm in which water and 10% sucrose water were always available, female rats drank about 200 cc of the sugar water (about 65 g of sucrose/kg) per day for 4 wk. There were no significant decreases in consumption over this time. In Exp. 2 female rats singly housed were given two bottles containing water for 1 wk. and then a bottle containing water and a bottle containing 15% sucrose for the next week for 6 wk. When sucrose was available, the rats ate 33% more rat chow. When sucrose was removed, the rats displayed more episodes of biting a stimulus when the food cubes were being removed for daily measurements. Some females exhibited a marked 4- to 5-day periodicity in sucrose (7.5%) consumption. The persistent and voluminous consumption of sucrose water and enhanced agonistic-like behavior during periods of withdrawal suggests the presence of a robust phenomenon with potential dinical applications to the challenge of addiction. PMID- 11899013 TI - Burnout among Iranian school principals. AB - This study investigated burnout among Iranian school principals. Also, the relationships of sex, years of administration, age, and marital status were considered. The sample were 200 principals (100 men, 100 women) who completed the Friedman School Principal Burnout Scale. Analysis showed principals who completed the scale felt exhausted, aloof, and deprecated. The women scored lower. There were significant correlationships between marital status and years of administration with the scores on burnout. PMID- 11899014 TI - A frequent misunderstanding associated with point biserial and phi coefficients. AB - The label "Pearson r" is legitimately used both for the standard Pearson r calculated on continuous variables and for its other varieties in the form of the point biserial or the phi coefficient. This fact is often ignored by psychologists and psychiatrists. PMID- 11899015 TI - Sociodemographic correlates of assimilation of refugees from Kurdistan. AB - Successful assimilation of refugees in their host country is an important prerequisite of psychological well-being. Refugees' satisfaction in the new country is one of key indicators of their assimilation. The satisfaction with their host country was assessed for 54 Kurdish refugees of mean age of 35.8 yr. (SD= 10.9) via an 8-item rating scale partly based on Cernovsky's Assimilation Scale. The 36 men and 18 women had resided in the host country for a mean of 4.5 yr. (SD=4.0). An overall score was calculated from ratings of satisfaction with personal safety, health, employment, food, financial security, social life, and entertainment. This overall score was unrelated to age, sex, and employment status. Those who emigrated at a younger age (r = -.28, p = .03) and those with lower education reported more satisfaction with their host country (r = -.28, p = .03) perhaps because they could more easily and rapidly re-establish social status comparable to what they had in their homeland than could older refugees from Kurdistan's higher educational strata. PMID- 11899016 TI - Children of homosexual parents report childhood difficulties. AB - Referenced as both supporting and weakening the case for parenting by homosexuals, 57 life-story narratives of children with homosexual parents published by Rafkin in 1990 and Saffron in 1996 were subjected to content analysis. Children mentioned one or more problems or concerns in 48 (92%) of 52 families. Of the 213 scored problems, 201 (94%) were attributed to the homosexual parent(s). Older daughters in at least 8 (27%) of 30 families and older sons in at least 2 (20%) of 10 families described themselves as homosexual or bisexual. These findings are inconsistent with propositions that children of homosexuals do not differ appreciably from those who live with married parents or that children of homosexuals are not more apt to engage in homosexuality. PMID- 11899017 TI - Acoustic determinants eliciting Japanese infants' vocal response to maternal speech. AB - Generally, infants prefer infant-directed speech to adult-directed speech. This study investigated which acoustic features of maternal infant-directed speech elicit effectively 3-mo.-old infants' vocal response. The participants were 40 Japanese mother and infant dyads. Vocal f0 from the mother's speech and the infant's vocalization was extracted using Computerized Speech Laboratory (CSL4300) and custom software. The acoustical features measured were mean fundamental frequency (f0), and f0 contour. The rate of the infant's vocal response was significantly higher When the maternal infant-directed speech was terminated with a falling contour rather than a rising or flat contour. There was no significant difference between the mean f0 of the maternal infant-directed speech followed or not followed by the infant's vocal response. This suggests that the falling contour of terminal maternal infant-directed speech serves to elicit the 3-mo.-old infant's vocal response. PMID- 11899018 TI - Cooperative learning and algebra performance of eighth grade students in United Arab Emirates. AB - This study investigated the effect of cooperative learning on eighth grade students' performance in algebra. 54 boys and 57 girls in four middle-school mathematics classes of Grade 8 in the UAE participated. Over a 3-wk. period, two classes (57 students) were taught using a cooperative learning method, and the other two classes (54 students) were taught using the traditional lecture method. Analysis of covariance using pretest scores as a covariant showed no statistically significant increase in the algebra performance for students in the cooperative learning groups compared with the traditional groups. However, boys in the cooperative setting improved significantly on the performance test compared with boys in the traditional setting. PMID- 11899019 TI - Lead, copper, zinc, and magnesium content in hair of children and young people with some neurological diseases. AB - The lead, copper, zinc, and magnesium levels of scalp hair taken from 153 children aged 1-15 yr and young people (16-18 yr) with selected neurological disorders (hyperexcibility, loss of consciousness, and epileptiform convulsions of an unknown origin, etc.), were measured using the atomic absorption spectrometry method and then compared with a control group of healthy children (n = 108). The research indicated significantly reduced mean levels of magnesium in the hair of children suffering from selected neurological diseases (in children aged 11-15 yr of age, above 30%; up to 5 yr of age, nearly 30%; the differences were statistically significant at p < 0.05) and slightly decreased mean levels of copper (differences statistically significant at p < 0.05, particularly in the 11 to 15-yr category). Differences in zinc levels in hair were inconsiderable (not statistically significant in any age groups). The lead level in the hair of the above-mentioned group of children was exceeded in relation to the control group (a statistically significant difference at p < 0.05 for the total group). A more than twofold decrease in the mean value of the Mg/Pb ratio (and a nearly 30% decrease in the value of the Mg/Zn ratio) in the hair of children suffering from neurological diseases suggests that the high toxicity of lead accompanying, among other things, magnesium deficiencies might be a cause of the observed disorders in children. PMID- 11899020 TI - Measurement of zinc, copper, manganese, and iron concentrations in hair of pituitary dwarfism patients using flameless atomic absorption spectrophotometry. AB - Pituitary dwarfism (hGHD) is known to be associated with trace element deficiency, which causes improper functioning of the involved endocrine system. Previously, we reported on the head hair concentrations of zinc, copper, manganese, and iron from a total of 418 normal subjects (154 male and 264 female). In this report, we analyzed the head hair concentrations of the same four trace metals of 103 hGHD children (60 male and 43 female) under treatment with human growth hormone (hGH). These subjects ranged in age from 5 to 18 yr. The results were compared with 338 age-matched normal subjects (120 male and 218 female). Both male and female hGHD showed approx 1.7 times higher zinc concentrations than normal subjects. Cheruvanky et al. reported a similar trend but with a slightly lower difference between hGHD and normal subjects. The average copper content in the hair of both male and female subjects also showed higher values for the hGHD than for the normal subjects, a trend similar to the values reported by Teraoka et al. In the case of manganese, the concentrations in hair of the hGHD were approx 50% of the values in the normal subjects. Head hair concentrations of iron in the hGHD were commensurate with the normal subjects. Because the content of trace elements in hair varies with the age of subjects, as a control, we investigated the head hair concentration of zinc from 20 healthy girls ranging in age from 10 to 18 yr. The average zinc concentration decreased from 10 to 12 yr, but no clear relation to age was observed from 13 yr and older. These trends were similar to our previous report. The zinc concentration in hair and body weight gain over a year was negatively correlated. The age variation in the content of zinc, copper, manganese, and iron in hair was measured comparing hGHD with the normal subjects in various ages. Concerning the zinc-level variation of hGHD and normal subjects, there were conspicuous differences between hGHD and normal subjects. For copper, the variations in concentration with age were similar to zinc. Regarding the age variations for manganese, hGHD had lower concentrations in hair compared to the normal subjects throughout adolescence (11 18 yr). We have studied the effects between the hair and these trace element concentrations in hGHD before and after hGH administration. These results suggest that hGH affects the metabolism of these trace elements. PMID- 11899021 TI - Dental amalgam affects urinary selenium excretion. AB - Selenium may have a protective effect against mercury toxicity. The aim of the present study was to investigate if selenium excretion in urine was affected in persons with dental amalgam fillings. The reason for this study is that dental amalgam is the most important source of inorganic mercury exposure in the general population, although the potential toxic effects of this exposure remain a subject for debate. The chelating agent 2,3 dimercaptopropane-1-sulfonate (DMPS) was injected intravenously (2 mg/kg) to provoke metal excretion. Urine samples were subsequently collected at intervals over a 24-h period. Selenium concentration was determined by hydride-generation atomic absorption spectrometry. The study was comprised of 20 persons who claimed symptoms from dental amalgam and 21 healthy persons with amalgam fillings. There were two control groups without amalgam. One control group had amalgam replaced because of concern about illness resulting from mercury release (n = 20), whereas the other control group never had amalgam (n = 19). Individuals with amalgam excreted less selenium (36.4 microg, median value) over 24 hours than those without amalgam (47.5 microg) (p = 0.016). There was no difference in selenium excretion between groups with (42.4 microg) and without (39.4 microg) amalgam-related symptoms (p = 0.15). The findings indicate that individuals exposed to low levels of elemental mercury from dental amalgam excrete less selenium to urine than unexposed individuals. PMID- 11899023 TI - Boron deficiency disables Xenopus laevis oocyte maturation events. AB - Processes of oocyte maturation that may be affected by boron (B) deficiency were studied to potentially determine a possible biochemical role of B in the Xenopus laevis oocyte. More specifically, the Xenopus oocyte membrane progesterone receptor (OMPR) in B-deficient oocytes was characterized by evaluating progesterone affinity for the OMPR and OMPR responsiveness to progesterone stimulation. The responsiveness of B-deficient oocytes to microinjection of a purified oocyte cytoplasmic fraction (OCF) from B-adequate oocytes was also studied to evaluate which aspects of the maturation process were affected by B deficiency. Results suggested that B deficiency resulted in incomplete oocyte maturation and that maturation could not be induced by the administration of exogenous progesterone. Progesterone successfully induced germinal vesicle breakdown (GVBD) in oocytes from females fed a B-supplemented diet (+ B) and females administered a traditional diet of beef liver and lung (B adequate). Addition of exogenous B to the -B oocytes increased the rate of progesterone induced GVBD slightly. The B-deficient X. laevis oocytes were capable of undergoing GVBD when endogenously stimulated by microinjected purified B-adequate OCF. These results indicated that the inability of the B-deficient oocytes to undergo GVBD was not associated with the cytoplasmic induction process specifically, but possibly in the progesterone receptor or signal transduction pathways. Radio-binding studies found that progesterone binding to the B deficient OPMR was greatly reduced compared to B-adequate or B-supplemented OMPR. Moreover, washout studies determined that progesterone binding to the OMPR in B deficient oocytes was more transient than the B adequate or +B oocytes. PMID- 11899022 TI - Study of the toxicological mechanism of acidified aerosols. AB - More than 30 aerosol samples were collected from 1 place near Zaojiaban Road (downtown of Shanghai) and 1 village near Jiading country (suburb) by a stacked filter air sampler. The coarse particulates (>2.5 microm) and the fine particulates (PM2.5) and their unsoluble parts were analyzed by proton-induced X ray emission. The cytotoxicity of particulates from the two places was evaluated by malondialdehyde (MDA) and thiazolyl blue (MTT) methods. The results show that the transition metal Fe, Cr, and Mn compounds from downtown are more easily soluble than those from the suburb, both in coarse and fine particulates, and the S content is much higher in particulates from downtown than that from the suburb. The cytotoxicity of the particulates from downtown is higher than that from the suburb and the cytotoxicity of acidified particulates is significantly higher than that of the controls. Because there are higher-soluble transition metal compounds that can induce free radicals in acidified particulates, the soluble transition metals may be one of the main factors for cytotoxicity. PMID- 11899024 TI - Effects of vanadate on leptin production from isolated rat adipocytes. AB - Vanadium, an important trace element in nutrition, actively affects the metabolic activities on all tissues. This work reports the effects of vanadate on leptin production from adipocytes isolated from normal Sprague-Dawley rats, streptozotocin-induced diabetic rats, Zucker lean, and Zucker fa/fa rats. We found that vanadate stimulates leptin production similar to insulin. The maximum leptin production stimulated by vanadate was slightly higher than that by insulin. Treatment with vanadate or insulin also increased the cellular leptin mRNA levels in a ratio similar to the increase in the protein secretion, suggesting that the regulation is at the transcription level. For adipocytes isolated from both streptozotocin-induced diabetic rats and Zucker fa/fa rats, vanadate was a more potent stimulant than insulin on leptin production. This suggests that vanadate may stimulate leptin production by mechanisms that are independent of insulin receptors. PMID- 11899025 TI - Inhibitory effects of phenylpropanoid metabolites on copper-induced protein oxidative modification of mice brain homogenate, in vitro. AB - We present the results of an in vitro investigation of the inhibitory effects of phenylpropanoid metabolites on copper-induced protein oxidative modification of mice brain homogenate. The effects of caffeic acid, 3-(3, 4-dihydroxyphenyl)-L alanine, esculetin, ferulic acid, and scopoletin were stronger than that of mannitol as a free-radical scavenger, whereas the effects of other phenylpropanoid metabolites, cinnamic acid, coniferyl alcohol, p-coumaric acid, coumarin, phenylalanine, tyrosine, and umbelliferone, were weak. These results demonstrated that phenolic carboxylic acids with 3,4-dihydroxy or 4-hydroxy-3 methoxy substituents and benzo-alpha-pyrons with 6,7-dihydroxy or 7-hydroxy-6 methoxy substituents in phenylpropanoid metabolites inhibit metal-induced protein oxidative modification of the brain. PMID- 11899026 TI - Stress hyperglycemia and diabetes mellitus in cats. PMID- 11899027 TI - Acute stress hyperglycemia in cats is associated with struggling and increased concentrations of lactate and norepinephrine. AB - We characterized the changes in blood glucose concentrations in healthy cats exposed to a short stressor and determined the associations between glucose concentrations, behavioral indicators of stress, and blood variables implicated in stress hyperglycemia (plasma glucose, lactate, insulin, glucagon, cortisol, epinephrine, and norepinephrine concentrations). Twenty healthy adult cats with normal glucose tolerance had a 5-minute spray bath. Struggling and vocalization were the most frequent behavioral responses. There was a strong relationship between struggling and concentrations of glucose and lactate. Glucose and lactate concentrations increased rapidly and significantly in all cats in response to bathing, with peak concentrations occurring at the end of the bath (glucose baseline 83 mg/dL, mean peak 162 mg/dL; lactate baseline 6.3 mg/dL, mean peak 64.0 mg/dL). Glucose response resolved within 90 minutes in 12 of the 20 cats. Changes in mean glucose concentrations were strongly correlated with changes in mean lactate (r = .84; P < .001) and mean norepinephrine concentrations (r = .81; P < .001). There was no significant correlation between changes in mean glucose concentrations and changes in mean insulin, glucagon, cortisol, or epinephrine concentrations. Struggling and lactate concentrations were predictive of hyperglycemia. Gluconeogenesis stimulated by lactate release is the likely mechanism for hyperglycemia in healthy cats in this model of acute stress. Careful handling techniques that minimize struggling associated with blood collection may reduce the incidence of stress hyperglycemia in cats. PMID- 11899028 TI - Development and evaluation of an enzyme-linked immunosorbent assay for the serodiagnosis of pythiosis in dogs. AB - Pythiosis (caused by the aquatic oomycete Pythium insidiosum) is a devastating and often fatal cause of either severe transmural gastroenteritis or locally invasive subcutaneous disease in dogs living in the southeastern United States. Although early diagnosis is essential for successful treatment, tools available for this task are limited. Therefore, we developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P insidiosum antibodies in canine serum. A soluble mycelial extract of P insidiosum was utilized as antigen in the ELISA, which was used to evaluate serum from 43 dogs with pythiosis, 8 dogs with lagenidiosis (another canine oomycosis), 16 dogs with nonoomycotic fungal or algal infections, 22 dogs with nonfungal gastrointestinal or skin disease, and 55 healthy dogs. Results were expressed as percent positivity (PP) relative to a strong positive control serum run on each plate. Medians and ranges for each of the 5 groups were as follows: pythiosis (81.7%, 50.6-98.5%), lagenidiosis (17.3%, 11.3-29.2%), other fungal or algal infections (8.2%, 4.7 15.4%), nonfungal gastrointestinal or skin disease (6.2%, 3.9-20.7%), and healthy dogs (6.7%, 3.0-15.2%). When using a cutoff value of 40% PP, the sensitivity and specificity of the ELISA both were 100%. In addition, ELISA values measured after successful surgical therapy in 2 dogs showed a decrease of anti-P insidiosum antibody concentrations into the normal range as early as 2 months after treatment. We conclude that the ELISA is a sensitive and specific test for the diagnosis of canine pythiosis, and may be a useful tool for monitoring response to medical or surgical therapy. PMID- 11899029 TI - Development of a nested polymerase chain reaction assay for the detection and identification of Pythium insidiosum. AB - Pythium insidiosum is an important cause of cutaneous and gastrointestinal disease in horses and dogs in the southeastern United States. Culture-based diagnosis of pythiosis is rarely definitive because production and identification of reproductive structures is difficult. The purpose of this study was to develop a polymerase chain reaction (PCR)-based assay for the identification of P insidiosum. Genomic DNA was extracted from 3 clinical isolates of P insidiosum and I isolate each of Pythium graminicola and Pythium arrhenomanes. The ITS I region of the ribosomal RNA gene of each isolate was amplified and sequenced, and the resultant sequences were aligned with published sequences for Pythium aphanidermatum, P acanthicum, and P myriotylum. A pair of P insidiosum-specific primers (PI-1 and PI-2) were designed from variable regions within the ITSI region. A nested PCR assay was developed in which the 1st round amplified the ITSI region by use of universal fungal primers. Second-round amplification utilized the internal P insidiosum-specific primers PI-1 and PI-2. Specificity of the assay was tested with DNA extracted from cultures of the following: 10 clinical isolates of P insidiosum and 1 isolate each of P graminicola, P irregulare, P arrhenomanes, P myriotylum, P deliense, Basidiobolus ranarum, Conidiobolus coronatus, Aspergillus terreus, Lagenidium giganteum, and a canine pathogenic Lagenidium species. Nested PCR produced a single 105-base pair amplicon for each of the P insidiosum isolates, but did not produce amplicons for any of the other isolates. Results of this study suggest that PCR is a useful tool for the identification of P insidiosum. PMID- 11899030 TI - Morphology of ventricular arrhythmias in the boxer as measured by 12-lead electrocardiography with pace-mapping comparison. AB - The QRS amplitude and polarity were determined in 12-lead electrocardiograms recorded from 22 Boxers with ventricular arrhythmias. Eighty-one percent (18/22) of dogs displayed a positive QRS morphology in the caudoventral leads (II, III, and aVF) and 77% (17/22) of dogs displayed a positive QRS morphology in the left precordial leads (V2-V6). In leads I and V1, the polarity of the QRS complex was variable (positive or negative). To determine if these morphologic features were suggestive of ventricular complexes arising from the right or left ventricle, a comparison was made to the QRS complexes in a pace-mapping study performed in 7 healthy mixed-breed dogs. A total of 3 right and 4 left ventricular sites were paced. None of the left ventricular paced sites resulted in a QRS morphology similar to the most common spontaneous ventricular arrhythmia in the Boxers. In contrast, QRS morphology in each of the 3 right ventricular sites was similar to that observed in the Boxers (P < .033). Each of these produced positive deflections in the caudoventral and left precordial leads, but both positive and negative QRS complexes were observed in leads I and V1 only when the right ventricular septum was paced. This finding suggested that the right ventricular septum might be a site of origin for the ventricular rhythm observed in the Boxers because in the Boxers the polarity of leads I and V1 also varied. Pacing the right ventricular outflow tract always resulted in a negative QRS complex in lead 1, whereas pacing the right ventricular apex always resulted in a positive QRS complex in lead I and a negative QRS complex in V1. However, these locations cannot be excluded as possible sites of origin for the spontaneous ventricular arrhythmias in the Boxers because the arrhythmias could be originating from both of these locations. The spontaneous ventricular arrhythmia of the Boxer is most similar to that of paced ventricular rhythms arising from the right ventricle. More precise localization to a region of the right ventricle such as outflow tract, septal, or apical could not be made. PMID- 11899031 TI - Bacterial meningoencephalomyelitis in dogs: a retrospective study of 23 cases (1990-1999). AB - The clinical records of 23 dogs (1990-1999) with histopathologically confirmed bacterial meningoencephalomyelitis were evaluated retrospectively. No breed, age, sex, or weight predisposition was found. All the dogs presented with clinical signs of a brain lesion, whereas 5 of 23 had neck pain. Pyrexia was detected in 11 of 23 dogs on admission. CBCs revealed neutrophilic leucocytosis in 7 of 21 dogs and thrombocytopenia in 3 of 21 dogs. The serum chemistry profiles were abnormal in 15 of 21 dogs. The results of cerebrospinal fluid (CSF) analysis were abnormal in 13 of 14 dogs and aerobic CSF culture was positive for bacteria in 1of 8 samples. At postmortem examination, the lesions were localized to the central nervous system. Escherichia coli, Streptococcus, and Klebsiella spp were the most frequently isolated bacteria from cultures collected at postmortem examination. Twelve papers reporting 51 total clinical cases of canine bacterial meningoencephalomyelitis were reviewed. The clinical signs and results of the CBC, serum chemistry, blood culture, and CSF analysis were collated and compared with those of this study. The results of the CSF analysis in this study were similar to those in the literature. CSF cultures documented in the literature were positive for Staphylococcus, Pasteurella. Actinomyces, Nocardia spp, and various anaerobic species including Peptostreptococcus, Eubacterium, and Bacteroides spp. PMID- 11899032 TI - Vascular endothelial growth factor concentrations in body cavity effusions in dogs. AB - Vascular endothelial growth factor (VEGF) has potent angiogenic, mitogenic, and vascular permeability enhancing properties specific for endothelial cells. VEGF is present in high concentrations in inflammatory and neoplastic body cavity effusions and has been implicated in the pathogenesis of neoplastic and inflammatory effusion formation. In this study, VEGF was quantitated by solid phase enzyme-linked immunoadsorbent assay (ELISA) in samples of pericardial, pleural, and peritoneal effusions (N = 38) from dogs (N = 35) with neoplastic and non-neoplastic diseases. VEGF was detected in 37 of 38 effusions (median, 754; range, 18-3,669 pg/mL) and was present in much higher concentrations than in previously established normal concentrations for canine plasma (median, < 1 pg/mL; range, < 1-18 pg/mL) or in those previously noted in the plasma of dogs with hemangiosarcoma (HSA; median, 17 pg/mL; range, < 1-67 pg/mL). In 4 dogs with HSA, the concurrent plasma VEGF concentration was much lower than in the abdominal effusion (P = .029). No significant correlation was demonstrated between VEGF effusion concentration and effusion total protein content or nucleated cell count. Mean VEGF concentrations were significantly higher in pericardial (median, 3,533; range, 709-3,669 pg/mL) and pleural effusions (median, 3,144; range, 0-3,663 pg/mL) compared to peritoneal effusions (median, 288; range, 18-2,607 pg/mL; P < .05). There was no marked difference demonstrated between effusions associated with malignant and nonmalignant diseases. Further studies are necessary to elucidate the role of VEGF in body cavity effusion formation in dogs. PMID- 11899033 TI - Idiopathic asymptomatic thrombocytopenia in Cavalier King Charles Spaniels is an autosomal recessive trait. AB - Cavalier King Charles Spaniels (CKCS) often have idiopathic asymptomatic thrombocytopenia. In affected dogs, the thrombocytes often are large, and it has been speculated that the condition could be an inherited macrothrombocytopenia. The aim of this study was to examine the inheritance of idiopathic, asymptomatic thrombocytopenia in CKCS. Sixteen families (both parents and > or = 3 offspring) of privately owned CKCS were included. There were 105 clinically healthy dogs (50 from Denmark and 55 from Sweden): 81 offspring and 26 parents (2 dogs had both roles). Because autoanalyzers have difficulty counting large platelets, the platelets were counted manually, with a counting chamber. Platelet counts were not influenced by age, gender, or heart murmur status. Thrombocytopenia (< or = 100,000 platelets/microL) was found in 46% of the parents. The pedigrees indicated that thrombocytopenia segregated as an autosomal recessive trait and that 100,000 platelets/microL was appropriate as a lower limit of normal. Affected offspring were found in all families, showing that all of the included parents were at least carriers. Therefore, the expected segregation ratios (which were in good accordance with the observed ones) were 1:0, 1:1, and 1:3 for the 3 crosses: affected x affected, normal x affected, and normal x normal. Within a given cross, the mean parental platelet count had no influence on the platelet counts of the offspring. We conclude that idiopathic, asymptomatic thrombocytopenia in CKCS is inherited in an autosomal recessive manner. The condition most likely constitutes an inherited macrothrombocytopenia in dogs. PMID- 11899034 TI - A retrospective study of 19 cases of canine myelofibrosis. AB - Nineteen cases of myelofibrosis were identified among 456 canine bone marrow specimens submitted for analysis. Myelofibrosis was classified as primary in I dog and as secondary in 18 dogs. Clinical conditions associated with secondary myelofibrosis included immune-mediated hemolytic anemia (n = 5), neoplasia (n = 4), and long-term drug treatment (n = 4). Drugs administered included phenobarbital, phenytoin, phenylbutazone, and colchicine. Bone marrow necrosis was observed in 5 dogs. Eight dogs were treated with immunosuppressive doses of prednisolone, and 3 were treated with erythropoietin. Half of the dogs with secondary myelofibrosis recovered from their cytopenias and were alive from 4 months to 5 years after diagnosis. PMID- 11899035 TI - Chemotherapy with cyclophosphamide, vincristine, and prednisolone (COP) in cats with malignant lymphoma: new results with an old protocol. AB - This retrospective study in 61 cats with malignant lymphomas examined the efficacy of a well-established chemotherapy protocol (cyclophosphamide, vincristine, and prednisolone [COP]) in the Netherlands, a country with a low prevalence of feline leukemia virus (FeLV). Twenty-two cats (36.1%) had mediastinal lymphoma, 11 (18.0%) had alimentary lymphoma, 7 (11.5%) had peripheral lymphoma, 8 (13.1%) had nasal lymphoma, and 13 (21.3%) had miscellaneous lymphoma (including renal lymphoma in 2 [3.3%]). Of the 54 cats that were tested, only 4 (7.4%) were FeLV positive. Complete remission (CR) was achieved in 46 of the 61 cats (75.4%). The estimated 1- and 2-year disease-free periods (DFPs) in the 46 cats with CR were 51.4 and 37.8%, respectively, whereas the median duration of remission was 251 days. The overall estimated 1-year survival rate in all cats was 48.7%, and the 2-year survival rate was 39.9%, with a median survival of 266 days. The median survival time and the 1-year survival rate for mediastinal lymphoma were 262 days and 49.4%. respectively. Siamese cats had a more favorable prognosis for survival and remission than other breeds. Response to therapy in this study was shown to be a significant prognostic indicator. CR is necessary for long-term survival. Cats that did not achieve CR had little chance of survival for longer than I year. Young Siamese cats in this study had a greater tendency to develop mediastinal malignant lymphoma at a young age, and all were FeLV negative. In comparison with results reported in other studies with different combination chemotherapy protocols, these are among the highest percentages of remission and the longest survival rates for cats with malignant lymphoma. PMID- 11899037 TI - Idiopathic vascular calcifications in a labrador retriever puppy. PMID- 11899036 TI - Serum immunoglobulin G concentrations in calves fed fresh colostrum or a colostrum supplement. AB - This study compared serum immunoglobulin G (IgG) concentrations in calves fed colostrum with those of calves fed a colostrum supplement containing spray-dried serum. Twenty-four Holstein calves were randomly assigned to I of 2 treatment groups (fresh colostrum or colostrum supplement). Each calf was fed 4 L of colostrum (n, = 12) or 4 L of colostrum supplement (n2 = 12) via oroesophageal intubation at 3 hours of age. The concentration of the colostrum supplement fed to calves was twice the manufacturer's recommendation. The median and range values for colostral IgG concentration were 6,430 mg/dL and 1,400-17,000 mg/ dL, respectively. Median serum IgG concentrations at 2 days of age differed significantly (P = .001) between calves receiving fresh colostrum (3,350 mg/dL) and the colostrum supplement (643 mg/dL). Eight percent of calves force fed colostrum had serum IgG concentrations < 1,000 mg/dL, whereas 75% of calves force fed supplement had IgG concentrations below this threshold. The calculated population relative risks for mortality associated with passive transfer for calves force-fed colostrum and calves force-fed colostrum supplement were 1.09 and 1.90, respectively. Force-fed fresh colostrum is superior to the colostrum supplement studied, but the colostrum supplement has similar efficacy to routine colostrum administration practices. PMID- 11899038 TI - Implantation of a single-lead atrioventricular synchronous (VDD) pacemaker in a dog with naturally occurring 3rd-degree atrioventricular block. PMID- 11899039 TI - Polyglucosan storage disease in a dog resembling Lafora's disease. PMID- 11899041 TI - Inactivation of Escherichia coli O157:H7 on inoculated alfalfa seeds with ozonated water and heat treatment. AB - Alfalfa seeds inoculated with a five-strain mixture of Escherichia coli O157:H7 were immersed in water containing 4, 8, 10, and 21 ppm of ozone for 2, 4, 8, 16, 32, and 64 min at 4 degrees C. Direct ozone sparging of seeds in water was used as an alternative mode of ozone treatment. Ozone-sparged seeds were also subsequently exposed to heat treatment at 40, 50, and 60 degrees C for 3 h. Populations of E. coli O157:H7 on untreated and treated seeds were determined by spread plating diluted samples on tryptic soy agar supplemented with 50 microg/ml of nalidixic acid. Since E. coli O157:H7 was released from inoculated seeds during treatment with ozone, the rate of release of cells from inoculated seeds soaked in 0.1% peptone water for up to 64 min was also determined. The overall reduction of E. coli O157:H7 on seeds treated with ozonated water without continuous sparging ranged from 0.40 to 1.75 log10 CFU/g (59.6 to 98.2%), whereas reductions for control seeds were 0.32 to 1.03 log10 CFU/g (51.7 to 90.5%). Treatment with higher ozone concentrations enhanced inactivation, but contact time longer than 8 min did not result in significantly higher reductions (P > 0.05). For seeds treated by ozone sparging, a 1.12-log10 CFU/g (92.1%) reduction was achieved using a 2-min contact time, and a 2.21-log10 CFU/g (99.4%) reduction was achieved with a 64-min contact time. The corresponding reductions for control seeds were 0.71 log10 CFU/g (79.5%) and 2.21 log10 CFU/g (99.4%), respectively. Treatment of ozone-sparged seeds at 60 degrees C for 3 h reduced the population to an undetectable level by direct plating (4 to 4.8 log10 CFU/g), although survivors were detected by enrichment. Ozone did not have a detrimental effect on seed germination percentage. PMID- 11899040 TI - Clinical syndrome associated with zolpidem ingestion in dogs: 33 cases (January 1998-July 2000). AB - Zolpidem is a nonbenzodiazepine hypnotic of the imidazopyridine class that is used to treat insomnia in humans. Zolpidem binds selectively to the benzodiazepine omega-1 receptor and increases the frequency of chloride channel opening, which results in inhibition of neuronal excitation. A retrospective study was conducted of zolpidem ingestion in dogs that were reported to the ASPCA Animal Poison Control Center (APCC) between January 1998 and July 2000. Data analysis included amount ingested, clinical effects, and time of onset of signs. Thirty-three reports of zolpidem ingestion in dogs (ranging in age from 5 months to 16 years) were evaluated. Approximate ingested dosages ranged from 0.24 to 21 mg/kg. Clinical signs reported included ataxia (18 dogs; 54.5%), hyperactivity (10 dogs; 30.3%), vomiting (7 dogs; 21.2%), and lethargy (5 dogs; 15.2%), as well as panting, disorientation, nonspecific behavior disorder, and hypersalivation (4 dogs each sign; 12.1%). Other signs reported include tachycardia, tremors, apprehension, vocalization, hypersalivation, weakness, and hyperesthesia. In 85% percent of reports, clinical signs developed within 1 hour and usually resolved within 12 hours. Although central nervous system (CNS) depression is reported as a primary effect of zolpidem in humans and would also be expected in dogs, information obtained from this study indicates that some dogs may exhibit a paradoxical excitation reaction. This effect appears to vary among individual dogs. PMID- 11899042 TI - Detection and elimination of Salmonella mbandaka from naturally contaminated alfalfa seed by treatment with heat or calcium hypochlorite. AB - In 1999, consumption of alfalfa sprouts contaminated with Salmonella Mbandaka led to a multistate outbreak of salmonellosis. In this study, the implicated alfalfa seed lot (no. 8,119) was confirmed to be contaminated with Salmonella Mbandaka at a detection frequency of approximately 72% per replicated 100 g of seed. The sensitivity of detection was improved by a combination of nonselective and selective enrichment of 5.0 ml of germination effluent, followed by immunomagnetic separation. Detection of low levels of viable cells with nonselective enrichment, employed to enhance the recovery of stressed or injured cells, was facilitated by the application of Salmonella-specific polymerase chain reaction (PCR). With PCR assays, Salmonella Mbandaka was detectable on seed stored at 5 degrees C for at least 11 months, but at an increasingly diminishing frequency. Using conventional techniques, viable populations were detected in the seed germination effluent from seeds stored for up to 8 months. Seed treatments with buffered (to pH 7) and unbuffered solutions of calcium hypochlorite, providing approximately 2,000 and 20,000 ppm of free chlorine, for 10 min were equally effective in eliminating viable populations of Salmonella Mbandaka. However, aqueous heat treatments at up to 85 degrees C for 1 min did not eliminate the naturally occurring contaminant from the seed. Reductions of > 15% in germination were observed following heat treatments of 65 degrees C for > or = 6 min or 70 degrees C for > or = 4 min. On the basis of these results, aqueous heat treatments alone do not appear to be a viable alternative to hyperchlorination as an effective method to eliminate Salmonella from alfalfa seed. PMID- 11899043 TI - Survival and growth of Listeria monocytogenes and Escherichia coli O157:H7 in ready-to-eat iceberg lettuce washed in warm chlorinated water. AB - Cut iceberg lettuce inoculated with Escherichia coli O157:H7 and Listeria monocytogenes before and after washing for 3 min in cold (4 degrees C) and warm (47 degrees C) water containing 100 mg/liter total chlorine was stored at I and 10 degrees C in oxygen-permeable film packages (6,000 to 8,000 cc/m2/24 h). Cold chlorinated water was detrimental to the survival of E. coli O157: H7 and L. monocytogenes at both storage temperatures. In contrast, washing in warm chlorinated water favored the growth of both pathogens in lettuce stored at 10 degrees C. There was no evidence of a relationship between the magnitude of spoilage microflora and the fate of either bacterium. PMID- 11899044 TI - Cross-contamination of lettuce with Escherichia coli O157:H7. AB - Contamination of produce by bacterial pathogens is an increasingly recognized problem. In March 1999, 72 patrons of a Nebraska restaurant were infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7, and shredded iceberg lettuce was implicated as the food source. We simulated the restaurant's lettuce preparation procedure to determine the extent of possible EHEC cross contamination and growth during handling. EHEC inoculation experiments were conducted to simulate the restaurant's cutting procedure and the subsequent storage of shredded lettuce in water in the refrigerator. All lettuce pieces were contaminated after 24 h of storage in inoculated water (2 x 10(9) CFU of EHEC per 3 liters of water) at room temperature or at 4 degrees C; EHEC levels associated with lettuce increased by > 1.5 logs on the second day of storage at 4 degrees C. All lettuce pieces were contaminated after 24 h of storage in water containing one inoculated lettuce piece (approximately 10(5) CFU of EHEC per lettuce piece) at both temperatures. The mixing of one inoculated dry lettuce piece with a large volume of dry lettuce, followed by storage at 4 degrees C or 25 degrees C for 20 h resulted in 100% contamination of the leaves tested. Microcolonies were observed on lettuce stored at 25 degrees C, while only single cells were seen on leaves stored at 4 degrees C, suggesting that bacterial growth had occurred at room temperature. Three water washes did not significantly decrease the number of contaminated leaves. Washing with 2,000 mg of calcium hypochlorite per liter significantly reduced the number of contaminated pieces but did not eliminate contamination on large numbers of leaves. Temperature abuse during storage at 25 degrees C for 20 h decreased the effectiveness of the calcium hypochlorite treatment, most likely because of bacterial growth during the storage period. These data indicate that storage of cut lettuce in water is not advisable and that strict attention must be paid to temperature control during the storage of cut lettuce. PMID- 11899045 TI - Prevalence of Escherichia coli associated with a cabbage crop inadvertently irrigated with partially treated sewage wastewater. AB - Preharvest contamination of field crops may have many sources, including feces, soil, and irrigation water. In March 2000, a sewage spill released unchlorinated tertiary-treated effluent into a creek used to irrigate commercial produce. A field of young cabbage transplants was irrigated with creek water as the contaminated water flowed past this land. Cabbage samples were taken from plots within this field, and Escherichia coli was isolated from the roots of these plants but not from the edible portion of the cabbage. No E. coli was isolated from water samples or from control samples taken from a nearby cabbage field watered with chlorinated municipal water. The cabbage field under study had not been fertilized with manure for at least 2 years prior to the contamination incident. Six different E. coli serotypes were identified, although none of them proved to be pathogenic. These serotypes were separated into five groups by a RiboPrinter; the resulting groups correlated well with the serotypes and the locations in the field from which these strains were isolated. We previously found that certain nonpathogenic E. coli strains displayed lower levels of adherence to lettuce seedling roots in a hydroponic adherence assay. The E. coli field strains displayed variable patterns of adherence to lettuce seedlings: strain MW421 showed significantly lower root and shoot adherence levels than did the other field strains, while strains MW423 and MW425 showed significantly higher root and shoot adherence levels. These data suggest that water quality is of paramount importance for the food safety of growing crops. PMID- 11899046 TI - Addition of fumaric acid and sodium benzoate as an alternative method to achieve a 5-log reduction of Escherichia coli O157:H7 populations in apple cider. AB - A study was conducted to develop a preservative treatment capable of the Food and Drug Administration-mandated 5-log reduction of Escherichia coli O157:H7 populations in apple cider. Unpreserved apple cider was treated with generally recognized as safe acidulants and preservatives before inoculation with E. coli O157:H7 in test tubes and subjected to mild heat treatments (25, 35, and 45 degrees C) followed by refrigerated storage (4 degrees C). Fumaric acid had significant (P < 0.05) bactericidal effect when added to cider at 0.10% (wt/vol) and adjusted to pH 3.3, but citric and malic acid had no effect. Strong linear correlation (R2 = 0.96) between increasing undissociated fumaric acid concentrations and increasing log reductions of E. coli O157:H7 in apple cider indicated the undissociated acid to be the bactericidal form. The treatment that achieved the 5-log reduction in three commercial ciders was the addition of fumaric acid (0.15%, wt/vol) and sodium benzoate (0.05%, wt/vol) followed by holding at 25 degrees C for 6 h before 24 h of refrigeration at 4 degrees C. Subsequent experiments revealed that the same preservatives added to cider in flasks resulted in a more than 5-log reduction in less than 5 and 2 h when held at 25 and 35 degrees C, respectively. The treatment also significantly (P < 0.05) reduced total aerobic counts in commercial ciders to populations less than those of pasteurized and raw ciders from the same source (after 5 and 21 days of refrigerated storage at 4 degrees C, respectively). Sensory evaluation of the same ciders revealed that consumers found the preservative-treated cider to be acceptable. PMID- 11899047 TI - Salmonella spp. shedding by alberta beef cattle and the detection of Salmonella spp. in ground beef. AB - Breeder cows, cattle recently arrived at feedlots, and cattle about to be shipped for slaughter were tested for Salmonella spp. No Salmonella spp. were detected in fecal samples from breeding cows. Nineteen of 1,000 (1.9%) fecal samples from recently arrived feedlot cattle were positive for Salmonella spp. compared to only 2 of 1,000 (0.2%) fecal samples taken within 2 weeks of slaughter. The positive fecal samples were collected in 5 of 50 (10%) "recent arrival" pens tested and in 1 of 50 (2%) pens tested within 2 weeks of slaughter. The serotypes isolated were Salmonella Agona, Salmonella Enteritidis, Salmonella Typhimurium DT104, and Salmonella 4,5,12:i:-. Ground beef samples purchased from retail outlets throughout Alberta were processed for Salmonella spp. Thirteen of 1,002 (1.3%) samples were positive for Salmonella spp. The serotypes isolated from ground beef were Salmonella Anatum, Salmonella Heidelberg, Salmonella Montevideo, Salmonella Typhimurium, Salmonella Typhimurium var. Copenhagen, and Salmonella Rough-O:i:1,2. The antibiotic resistance and pulsed-field electrophoresis gel macrorestriction patterns of all isolates were compared. PMID- 11899048 TI - Adhesion and colonization of Vibrio cholerae O1 on shrimp and crab carapaces. AB - The potential of Vibrio cholerae O1 to attach to and colonize the carapaces of shrimp and crabs was evaluated. One million cells of V. cholerae O1 were spread within a circle on the external surfaces of separated carapaces and stored at 22 +/- 0.2 degrees C in a moist environment to permit adherence. Attached vibrios were counted directly by an immunofluorescence technique and by the pour plate technique after detachment of the cells. To study the colonization process, rifampicin-resistant strains of V. cholerae O1 were used. V. cholerae O1 strains, including those resistant to rifampicin, were able to attach to shrimp and crab carapaces. Dorsal crab carapaces showed higher levels of attachment than ventral carapaces. Colonization of V. cholerae O1 on these carapaces was also demonstrated. Both attachment and colonization on the shrimp exoskeleton were optimal at a salinity of 1.0 to 1.5%, a pH of 6.0 to 7.0, and a temperature of 37 degrees C. Less than 2% attachment at 3 degrees C contrasted with >20% attachment at 37 degrees C. Even at 3% NaCl, some attachment was observed. Although attachment percentages may appear low (2 to 20%), they represent significant numbers, about 3.7 to 5.6 log10 CFU per carapace. A rugose V. cholerae O1 strain attached to and colonized the shrimp carapace in a fashion very similar to that of the smooth strain from which it was derived. The ability of V. cholerae O1 to attach to and colonize exoskeletons of edible crustaceans provides a potential means of survival in aquatic environments. Concentrations of vibrios that may be reached on a single crab or shrimp carapace are clearly of concern with regard to public health. PMID- 11899049 TI - Effects of heat shock on the thermotolerance, protein composition, and toxin production of Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus, an important seafood-associated enteropathogen, usually encounters different adverse conditions in its native or food-processing environment, and the stresses resulting from these conditions may affect the survival of this pathogen and thus change its risk with regard to food hygiene. In this study, we investigated the thermotolerance of V. parahaemolyticus under sublethal heat shock and characterized this response by examining the changes in protein profiles and toxin production. Logarithmically grown cells heat shocked at 42 degrees C for 30 min were more resistant to thermal inactivation at 47 degrees C than were unshocked cells. After the 25 degrees C culture was heat shocked, 24 species of proteins were induced, while 13 species were inhibited, as indicated by polyacrylamide gel electrophoresis. DnaJ-, GroEL-, and GroES-like proteins with molecular sizes of 47, 62, and 12 kDa, respectively, were detected by immunoblotting with antibodies raised against the Escherichia coli proteins. During 1 to 8 h of heat shock, GroEL-like protein was produced in substantial amounts and was present in the periplasmic and extracellular fractions, while DnaJ- and GroES-like proteins were present mainly in the total cellular fraction. DnaK-like protein was not detected; nevertheless, the presence of the dnaK-like genetic element was revealed by Southern blotting. Production of thermostable direct hemolysin, the major virulence factor in V. parahaemolyticus, was enhanced in the cells heat shocked at 42 degrees C but not in those heat shocked at 37 degrees C. PMID- 11899050 TI - Control of natural microbial flora and Listeria monocytogenes in vacuum-packaged trout at 4 and 10 degrees C using irradiation. AB - The effect of gamma irradiation on the natural microflora of whole salted vacuum packaged trout at 4 and 10 degrees C was studied. In addition, the effectiveness of gamma irradiation in controlling Listeria monocytogenes inoculated into trout was investigated. Irradiation at doses of 0.5 and 2 kGy affected populations of bacteria, namely, Pseudomonas spp., Brochothrix thermosfacta, lactic acid bacteria, H2S-producing bacteria typical of Shewanella putrefaciens, and Enterobacteriaceae, at both 4 and 10 degrees C. This effect was more pronounced at the higher dose (2 kGy) and the lower temperature (4 degrees C). Pseudomonads, H2S-producing bacteria typical of S. putrefaciens, and Enterobacteriaceae showed higher sensitivity to gamma irradiation than did the rest of the microbial species. Sensory evaluation did not show a good correlation with bacterial populations. On the basis of sensory odor scores, a shelf life of 28 days (2 kGy, 4 degrees C) was obtained for salted vacuum-packaged freshwater trout, compared with a shelf life of 7 days for the unirradiated sample. Under the same conditions, the growth of L. monocytogenes inoculated into the samples was suppressed by 2 log cycles after irradiation (2 kGy) and storage for up to 18 days at 4 degrees C. PMID- 11899051 TI - Effects of diacetyl and carbon dioxide on spoilage microflora in ground beef. AB - The effect of CO2 and diacetyl, alone or in combination, on spoilage microflora in ground beef was determined. Ground beef was treated with 20, 30, or 40% CO2 for 22 days (study I); 20, 50, or 100 microg/g diacetyl for 26 days (study II); or a combination of 20% CO2 and 100 microglg diacetyl for 40 days (study III). Antimicrobial effectiveness was determined by aerobic plate counts (log10 CFU/g) using plate count agar (total aerobic bacteria), deMan Rogosa Sharpe (MRS) Lactobacillus agar (gram-positive bacteria), MacConkey agar (gram-negative bacteria), pH, and informal organoleptic assessments (by appearance and by odor). In study I, total bacteria and pH increased by day 4 in control meat samples. For all CO2 levels, gram-negative bacteria decreased and gram-positive bacteria increased compared with untreated controls. The pH remained constant for CO2 treated meat. Control samples had an off-odor and a brown appearance, while CO2 treated samples had no off-odor but did have a brown appearance. For samples treated with diacetyl (study II), spoilage was evident by day 7 for samples treated with 0, 20. and 50 microg/g diacetyl for all parameters examined. Ground beef treated with 100 microg/g diacetyl was spoiled on day 12. Diacetyl was detected (by odor) in samples that were treated with 100 microg/g diacetyl and had a brown appearance. Meat samples treated with the combination of CO2 and diacetyl (study III) showed that the addition of diacetyl did not have an additive effect on microbial growth. Combination-treated meat maintained a red appearance and no off-odor. Diacetyl and CO2 could be used in combination to maintain a red color and inhibit spoilage microorganisms. PMID- 11899052 TI - Inhibition, resistance development, and increased antibiotic and antimicrobial resistance caused by nutraceuticals. AB - With the increasing use of herbal remedies by the general public worldwide, there remains a lack of information on the relationship between nutraceutical use and antibiotic resistance. Historically, there have been claims that nutraceuticals possess antibacterial and antiviral activity. However, the claims come with little or no documentation and no information related to the development of resistance to the nutraceutical or the cause of increases in resistance to antibiotics. These studies investigate the ability of nutraceutical exposure to influence the development of antibiotic resistance in bacteria. Two antibiotic sensitive organisms, Staphylococcus aureus ATCC 29213 and Escherichia coli ATCC 25922, were used as representative of the gram-positive and gram-negative bacteria. These preliminary investigations showed a general increase to the ampicillin marker by the products studied, using Staphylococcus aureus ATCC 29213 as the indicator organism. There were 13 product-related increases in the MIC, 2 decreases, and 7 no changes. All six of the garlic products increased the MIC of the norfloxacin marker to greater than fourfold above baseline. Using E. coli ATCC 25922 as the indicator organism, the greatest product-antibiotic marker interaction was with the ampicillin marker. Garlic, Echinacea, and zinc products all caused large increases in the MIC to ampicillin over baseline values. PMID- 11899053 TI - Adhesion of dairy propionibacteria to intestinal epithelial tissue in vitro and in vivo. AB - Adhesion to the intestinal mucosa is a desirable property for probiotic microorganisms and has been related to many of their health benefits. In the present study, 24 dairy Propionibacterium strains were assessed with regard to their hydrophobic characteristics and their autoaggregation and hemagglutination abilities, since these traits have been shown to be indicative of adherence in other microorganisms. Six strains were further tested for their capacity to adhere to ileal epithelial cells in vitro and in vivo. The results of the study showed that propionibacteria were highly hydrophilic, and hemagglutination and autoaggregation were properties not commonly found among these microorganisms. No relationship was found between surface characteristics and adhesion ability, since hemagglutinating, autoaggregating, and nonautoaggregating bacteria were able to adhere to intestinal cells both in vitro and in vivo. Microscopic examination revealed that autoaggregating cells adhered in clusters, with adhesion being mediated by only a few bacteria, whereas the hemagglutinating and nonautoaggregating strains adhered individually or in small groups making contact with each epithelial cell with the entire bacterial surface. The in vitro assessment of adhesion was a good indication of the in vivo association of propionibacteria with the intestinal epithelium. Therefore, the in vitro method presented here should be valuable in screening routinely adhesive properties of propionibacteria for probiotic purposes. The adhesion ability of dairy propionibacteria would prolong their maintenance in the gut and increase the duration of their provision of beneficial effects in the host, supporting the potential of Propionibacterium in the development of new probiotic products. PMID- 11899054 TI - Yersinia pseudotuberculosis with limited genetic diversity is a common finding in tonsils of fattening pigs. AB - A total of 425 pig tonsils, including 210 tonsils from fattening pigs and 215 from sows, from seven different abattoirs in Finland were studied for the occurrence of Yersinia pseudotuberculosis from 1999 to 2000. The mean prevalence of Y. pseudotuberculosis in fattening pig tonsils was 4%, varying from 0 to 10% between slaughterhouses. Y. pseudotuberculosis was not recovered from sow tonsils. All 30 Y. pseudotuberculosis isolates from eight pig tonsils were recovered after cold enrichment. Seventeen isolates from seven tonsils were found after cold enrichment for 14 days, followed by alkali treatment. Y. pseudotuberculosis was not isolated after direct plating, overnight enrichment, or selective enrichment. All 30 isolates belonged to bioserotype 2/0:3 and carried the virF gene in the virulence plasmid. The isolates exhibited calcium dependence and Congo red absorption. The pyrazinamidase test gave variable results. All isolates were characterized with pulsed-field gel electrophoresis (PFGE). Using SpeI, NotI, and XbaI enzymes, seven, five, and two different PFGE patterns were obtained, respectively. A total of 11 genotypes, gI to gXI, identified by a combination of the various SpeI, NotI, and XbaI profiles, were detected. Three pigs were found to carry more than one genotype. Overall, variations between PFGE patterns were small, indicating genetic homogeneity among pig strains of bioserotype 2/0:3. PMID- 11899055 TI - Modification of Niven's medium for the enumeration of histamine-forming bacteria and discussion of the parameters associated with its use. AB - The objective of this study was to improve Niven's medium (NM) for the optimized enumeration of histamine-forming bacteria (HFB). The parameters modified related to solidification of the agar at low pH values (pH 5.3 to 5.8), incubation time (24, 48, and 72 h) and temperature (30 and 37 degrees C), number of colonies developed on the plate to allow enumeration of HFB, and color differentiation. Strains of HFB, Morganella morganii, Klebsiella pneumoniae, and Hafnia alvei were examined for their ability to change color on NM. The three microorganisms produced different colors on the medium, which can be used for preliminary identification of HFB. Quantitative analysis of HFB proved to be achievable, with the prerequisite that only 1 to 80 colonies developed on the medium allow effective enumeration. A larger number of colonies results in color development throughout the medium, making the distinction between HFB and other bacteria unachievable. Growth of prolific HFB was noticeably better at pH values from 5.3 to 5.5, compared to 6.3, on NM. Growth at 5.3 and 5.5 on NM also presented a significant advantage in comparison to growth on plate count agar (PCA; pH 7) at the same incubation temperature. The increased agar concentration of 3% was found to give better solidification at pH 5.3 to 6.0, compared to 2%. This agar concentration also allows autoclaving for 12 min at 121 degrees C, overcoming the hydrolysis problems that appear at the lower concentration of 2%. The construction of a color chart for the recognition of the pH change due to histidine decarboxylase activity was also achieved. PMID- 11899056 TI - Detection of Salmonella in foods using Tecra Salmonella VIA and Tecra Salmonella UNIQUE rapid immunoassays and a cultural procedure. AB - The presence of Salmonella in 200 raw food samples of animal origin was investigated by means of the rapid immunoassays Tecra Salmonella VIA and Tecra Salmonella UNIQUE (Tecra Diagnostics, Rosewille, New South Wales, Australia) and a cultural procedure. Forty-five samples (22.5%) were Salmonella positive by at least one of the three methods. The number of positive samples according to the analytical method was 34 (75.6%) for the cultural procedure, 29 (64.4%) for Tecra Salmonella VIA, and 27 (60.0%) for Tecra Salmonella UNIQUE. Tecra Salmonella UNIQUE detected three positive samples that were not detected by the two other methods. The cultural method also detected three positive samples that both rapid methods were unable to detect. McNemar's chi-square tests indicated that the differences between results given by the rapid immunoassays when compared with those of the cultural method were not significant (P > 0.05). PMID- 11899057 TI - Radiation resistance of virulence plasmid-containing and plasmid-less Yersinia enterocolitica. AB - Yersinia enterocolitica, a foodborne pathogen, can be eliminated from meat by ionizing radiation. Y. enterocolitica sometimes contains a 70-kb virulence plasmid that encodes genes for a type III secretion channel and host immune suppression factors. The radiation resistance of virulence plasmid-containing and plasmid-less Y. enterocolitica was determined. Four Y. enterocolitica serotypes containing (i) the large virulence plasmid, and (ii) their plasmid-less derivatives were inoculated into raw ground pork, which was then vacuum packed and irradiated at 4 degrees C to doses of 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. The D10-value, the radiation dose required to reduce the number of viable Y. enterocolitica by 90%, was not dependent on the presence or absence of the virulence plasmid, but it did differ among the four Y. enterocolitica serotypes. PMID- 11899058 TI - Effects of refrigeration and alcohol on the load of Aeromonas hydrophila in oysters. AB - Members of the bacterial genus Aeromonas are widely distributed throughout the environment and are readily cultured from a variety of foods. One member of this genus, Aeromonas hydrophila, has been reputed to be a significant cause of gastrointestinal disease. In this study, we examined the effects of refrigeration and alcohol on the level of A. hydrophila in oysters. Specifically, vodka was examined because it is used by the food service industry in preparation of Oysters Romanoff. One set of oysters was shucked on receipt, whereas others were refrigerated intact for 7 days at 5 degrees C. The oysters were blended and the numbers of A. hydrophila present determined using starch ampicillin agar. Oysters were also shucked and placed on the half shell with 5 ml of vodka for 10 min. The oysters were then washed and presumptive A. hydrophila levels determined in both the washate and homogenate. On the day of purchase, the average number of presumptive A. hydrophila found was 7.6 x 10(4) CFU/g of oyster meat. After 7 days of refrigeration, the average number had increased to 3.2 x 10(5) CFU/g of oyster meat. In the oysters treated with vodka, the average number of A. hydrophila present internally was 9.9 x 10(4) with high numbers (10(3) to 10(4)) isolated from the oyster surface. From these data, it is clear that refrigeration and alcohol treatment are not sufficient to reduce loads of A. hydrophila in or on oysters. PMID- 11899059 TI - Determination of MICs of streptomycin for resistant Salmonella isolates in swine and poultry using a micro-broth dilution system. AB - The MICs of streptomycin for Salmonella isolates from swine and poultry were determined by a micro-broth dilution technique. The Salmonella isolates were recovered from the lymph nodes and cecal contents of market-age swine and from the cecal contents of poultry at the time of slaughter and were found by disk diffusion to be resistant to 10 microg of streptomycin. MIC testing was carried out with the Sensititre susceptibility system for streptomycin, which uses a microwell concentration gradient of 16 to 800 microg/ml. Results indicated that >80% of the swine isolates had MICs of < or = 64 microg/ml, while 51% of poultry isolates exhibited MICs of > or = 128 microg/ml. The highest MICs observed in swine and poultry were 256 and 800 microg/ml, respectively. Replicate tests performed on 12 of the isolates chosen at random indicated a 100% correlation between runs. Advantages of this system include easily read results and precoated wells. Disadvantages include the cost and the inability to test concentrations of streptomycin other than those in the wells. We found this micro-broth dilution commercial test kit to provide a relatively quick and easy testing procedure for the determination of streptomycin resistance in Salmonella. PMID- 11899060 TI - Reduction of spoilage microorganisms in fresh beef using hydrodynamic pressure processing. AB - Hydrodynamic pressure processing (HDP) was investigated as a technology to reduce spoilage microorganisms found in fresh beef. In two separate studies (studies 1 and 2), retail ground beef and beef roasts were purchased (day 0). The roasts were divided into stew pieces (30 to 40 g). All meat samples, including control samples, were stored at 5 degrees C for 20 h in a plastic film. After storage, designated samples were treated with HDP In study 3, ground beef was treated with HDP (day 0) and stored aerobically (5 degrees C) for 14 days with control samples. Each meat type was vacuum-packaged for HDP (100 g binary explosive, steel shock wave container). The pHs and the aerobic plate counts (log10 CFU/g) were measured on day 0 (studies I and 2) and on days 0, 7, and 14 (study 3) for control samples and for HDP-treated samples. There was no pH difference between control and HDP-treated meat types (studies 1 and 2); HDP reduced bacteria in both meat types in study 1 (2 log) and study 2 (1.5 log) on day 0. In study 3, there was a significant difference (P < 0.05) in pH between control meat (8.2) and HDP-treated meat (5.6) after storage. There was an immediate reduction (1.5 log) of microorganisms following HDP (day 0) and a 4.5-log difference between control samples (9 log) and HDP-treated samples (4.5) after 14 days of storage. With HDP, it is possible to reduce spoilage microorganisms found in or on different meat types (ground beef versus stew pieces), which could extend the shelf life of meat products. PMID- 11899061 TI - Aeromonas species in foods. AB - Aeromonas species have been recognized as potential or emerging foodborne pathogens for more than 20 years. Aeromonads are estuarine bacteria and are ubiquitous in fresh water, fish and shellfish, meats, and fresh vegetables. Actual sourced foodborne outbreaks are few, but epidemiological evidence suggests that the bacterium can cause self-limiting diarrhea, with children being the most susceptible population. Most aeromonads are psychrotrophic and can grow in foods during cold storage. Aeromonads are not resistant to food processing regimes and are readily killed by heat treatment. A host of virulence factors are present, but the exact role of each in human disease has not been fully elucidated. PMID- 11899062 TI - Miniaturized formats for efficient mass spectrometry-based proteomics and therapeutic development. AB - Off-line miniaturized "nano-spray" formats for electrospray ionization mass spectrometry (ESI-MS) enable the routine identification of femtomole quantities of protein or peptide. Even greater strides have been achieved using on-line miniaturized ESI-MS methods, such as nanobore LC-MS and CE-MS. On-line methods enable greater sensitivity (sub-attomole limit of detection), dynamic range, and throughput. In either off- or on-line methods for protein analysis, samples are typically isolated and digested enzymatically, with MS analysis of the peptide fragments, yielding 5-50% sequence coverage, in a "bottom-up" approach. Obtaining biologically relevant (structure/function) information (such as the localization of regions of error or post-transnational modifications) often demands 100% sequence coverage and this may be obtained by analyzing intact proteins by MS with a "top-down" methodology. Proteome wide success with top-down methods will require the development of novel miniaturized approaches for sample preparation along with new tools for bioinformatics. As these miniaturized formats continue to power proteomics applications, they will undoubtedly pollinate "cross-over" applications in LC-MS ranging from drug discovery to development. An example of metabolite identification using an order of magnitude less sample than usually required, with a concurrent order of magnitude increase in signal, illustrates the potential of miniaturized formats in lead characterization activities. PMID- 11899063 TI - Early discovery drug screening using mass spectrometry. AB - Electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometric methods useful for early discovery drug screening are reviewed. All methods described involve studies of non-covalent complexes between biopolymer receptors and small molecule ligands formed in the condensed phase. The complexes can be sprayed intact directly into the gas phase by ESI-MS using gentle experimental conditions. Gas phase screening applications are illustrated for drug ligand candidates non-covalently interacting with peptides, proteins, RNA, and DNA. In the condensed phase, the complexes can be also isolated, denatured and analyzed by ESI-MS to identify the small molecule ligands. Condensed phase drug screening examples are illustrated for the ESI-MS ancillary techniques of affinity chromatography, ultrafiltration, ultracentrifugation, gel permeation chromatography (GPC), reverse phase-high performance liquid chromatography (RP-HPLC) and capillary electrophoretic methods. Solid phase drug screening using MALDI-MS is illustrated for small molecule ligands bound to MALDI affinity probe tips and to beads. Since ESI and MALDI principally produce molecular ions, high throughput screening is achieved by analyzing mass indexed mixtures. PMID- 11899064 TI - Metabonomic applications in toxicity screening and disease diagnosis. AB - Biofluid NMR spectroscopy is a powerful tool providing a comprehensive metabolic profile of the low molecular weight components in biofluids that reflect concentrations and fluxes of endogenous metabolites involved in key intermediary cellular pathways, thereby giving an indication of an organisms physiological or pathophysiological status [1]. The interaction of pharmacological agents with cells and tissues can also be monitored using recently developed high resolution magic-angle spinning (HRMAS) NMR spectroscopic technology for biological matrices [1]. However, recent developments in both spectrometer and software technology has resulted in improved capacity for sample handling, leading to a rapid growth in the size of toxicological spectral databases, and increased the complexity of the biological spectral data generated. Thus more emphasis has been placed on the need to develop improved automated procedures for data processing and interpretation. By harnessing chemometric tools for analysis of complex spectral data, the toxicological consequences of xenobiotic exposure can be evaluated efficiently on line. Automation of spectral processing procedures and the construction of mathematically based 'expert systems' for the prediction of drug induced toxicity founded on IH NMR spectral profiles have now been achieved. Chemometric analysis of biological NMR spectra has provided the main analytical platform for metabonomic analysis, providing a systems approach to evaluating pathophysiological or genetic influences on the metabolic status of an organism [1]. This technology is currently being given high-priority in the pharmaceutical industry with respect to development of efficient high throughput toxicity screening systems for lead candidate selection. In this article, we review the recent developments in metabonomics and consider their application in toxicological screening, disease diagnosis and functional genomics. PMID- 11899065 TI - Recent advances in use of LC/MS/MS for quantitative high-throughput bioanalytical support of drug discovery. AB - LC/MS/MS based bioanalysis using atmospheric pressure ionization (API)-style interfaces has now been applied for over a decade. This technology, which initially found application for clinical bioanalysis, is now firmly established as the primary bioanalytical tool for ADME studies related to drug discovery and lead optimization (LO). This review focuses on recent advances in LC/MS/MS based bioanalysis in support of drug discovery and LO. The initial part of the article reviews the principal components of LC/MS/MS bioanalysis: sample preparation, chromatography, ionization and mass analysis. In each section, factors affecting high throughput bioanalysis are addressed. Because of the importance of on-line column switching methods to discovery bioanalysis, the section on sample preparation is divided into off-line and on-line approaches. In addition, the discussion of chromatography is limited to reversed phase liquid chromatography with emphasis given to the trend towards high-flow gradient elution techniques. The latter part of the review focuses on considerations for experimental design. In this section, pooling methods such as cassette dosing are discussed along with more highly integrated strategies linking bioanalysis with protocol generation and sample collection. The article concludes by briefly reviewing factors, which affect bioanalytical precision and accuracy, such as ion suppression, analyte stability and metabolite interference. PMID- 11899066 TI - Organ perfusion and mass spectrometry: a timely merger for drug development. AB - Organ perfusion techniques bridge the methodological gap between in vivo studies on the one hand and in vitro studies on the other. In drug candidate selection and subsequent development the differences between these systems should be considered carefully in study design, as one approach may be more suitable than the other depending on the question(s) being asked and, in particular, how the data will be used. This article is not concerned with the mechanics, the surgery or composition of perfusates as there are numerous reviews/books covering these aspects. Instead, using perfused gut, liver, lung, kidney and brain as examples, the emphasis is on the usefulness (or otherwise) of the data generated with respect to drug absorption, metabolism, pharmacokinetics (PK) and the factors which affect these parameters. Perfusion systems are not difficult to set up but do require 'high maintenance' for routine use. For this reason they have been used sparingly by the pharmaceutical industry mainly for problem solving or mechanistic studies. The latter part of this article shows how simultaneous dosing of numerous compounds followed by multiple--component analysis using LC/MS/MS has proved to be an effective way to improve the throughput of absorption, pharmacokinetics and metabolism screening ex vivo. PMID- 11899067 TI - Multivariate pharmaceutical profiling for drug discovery. AB - The field of pharmaceutical profiling in drug discovery is described. The pharmaceutical properties of drug candidates determine how much of the drug safely reaches the therapeutic target. Drug candidates often fail in discovery and development due to inadequate properties, resulting in lost opportunities and resources for developing new drugs. Pharmaceutical profiling assays have been developed and implemented to measure the properties of large numbers of drug candidates starting at the earliest stages of discovery. This information is used for informed decisions in drug candidate selection and synthetic optimization. A holistic process of parallel activity and property optimization has emerged in drug discovery. The assays, strategies, and data management associated with pharmaceutical profiling are discussed. PMID- 11899068 TI - Applications of computer software for the interpretation and management of mass spectrometry data in pharmaceutical science. AB - The rapid growth of mass spectrometry (MS)-based computer software applications has been fueled by the unprecedented need to capture and analyze MS data and provide the information necessary for decision-making. Shorter timelines and a significantly greater number of samples has resulted in a tremendous focus on streamlined approaches that provide scientists, managers, and executives the capability to readily obtain, or even request, the necessary information that leads to accelerated product development. The generation of analytical data using roboticized high-throughput hardware has produced a bottleneck since data can be generated faster than it can be analyzed. New techniques including MS/MS and accurate mass experiments are feasible only using computers to capture and manage the enormous amounts of data necessary to perform the experiments. Whatever the nature of the experiments conducted, the MS analysis strategy is to extract the appropriate information required for decision-making in as facile a manner as possible. We will review here a survey of the creation of commercial and laboratory specific reference databases and associated searching algorithms and also recent efforts to introduce advancedprocessing and analysis algorithms to the hands of the masses, specifically as an aid to structure elucidation. PMID- 11899069 TI - Measles virus induced immunosuppression: targets and effector mechanisms. AB - A profound, transient suppression of immune functions during and after the acute infection is the major cause of more than one million cases of infant deaths associated with measles worldwide. Concommittant with the generation of an efficient measles virus (MV) specific immunity, immune responses towards other pathogens are strongly impaired and provide the basis for the establishment and severe course of opportunistic infections. The molecular basis for MV-induced immunosuppression has not been resolved as yet. Similar to other immunosuppressive viruses, MV is lymphotropic and viral nucleic acid and proteins are detectable in peripheral blood mononuclear cells (PBMC). It is considered central to MV-induced immunosuppression that PBMC isolated from patients largely fail to proliferate in response to antigen specific and polyclonal stimulation. The low abundancy of MV-infected PBMC suggests that MV-induced immunosuppression is not directly caused by infection-mediated cell loss or fusion, but rather by indirect mechanisms such as deregulation of cytokines or surface contact-mediated signaling which may lead to apoptosis or impair the proliferative response of uninfected PBMC. Evidence for a role of any of these mechanisms was obtained in vitro, however, much has still to be learned about the tropism of MV and its interactions with particular host cells such as dendritic cells in vivo. PMID- 11899070 TI - The molecular pathogenesis and experimental therapy of IgA nephropathy: recent advances and future directions. AB - In 1968 Berger and Hinglais published the first description of IgA nephropathy (IgAN). In the ensuing 30 years, extensive clinical, epidemiologic, and immunologic characterizations of primary (idiopathic) glomerulonephritis with IgA as the predominant or co-dominant immunoglobulin deposited in the mesangia of all glomeruli, have established the features of IgAN as a distinct glomerular disease entity. Despite these efforts, the basic molecular mechanism(s) which mediate abnormal mesangial IgA deposition with ensuing extracellular matrix expansion and mesangial cell proliferation remains poorly understood, definitive diagnosis still depends on histologic examination of renal biopsy specimens, and widely accepted standards for effective therapy remain to be defined. This review will begin with a summary of the earlier 'descriptive' histopathologic and clinical epidemiologic work which firmly established the distinct immunohistologic features of IgAN, the most common glomerulonephritis among patients undergoing renal biopsy and a major cause of renal failure worldwide. In recent years, a series of important advances in the areas of molecular pathogenesis and experimental therapy have emerged, reflected in a "molecular" paradigm shift in the techniques and approaches applied to the study of IgAN. Representative studies will be critically evaluated to highlight both the strengths and potential weaknesses of each of these approaches. Throughout, the author will offer a personal perspective on promising areas of new investigation and potential approaches to the identification of disease/susceptibility genes involved in the development and progression of IgAN, the application of these discoveries through the development of clinically useful molecular diagnostic tests, and the rational design of novel therapeutic strategies. PMID- 11899071 TI - Molecular mechanisms of thiamine utilization. AB - Thiamine is required for all tissues and is found in high concentrations in skeletal muscle, heart, liver, kidneys and brain. A state of severe depletion is seen in patients on a strict thiamine-deficient diet in 18 days, but the most common cause of thiamine deficiency in affluent countries is alcoholism. Thiamine diphosphate is the active form of thiamine, and it serves as a cofactor for several enzymes involved primarily in carbohydrate catabolism. The enzymes are important in the biosynthesis of a number of cell constituents, including neurotransmitters, and for the production of reducing equivalents used in oxidant stress defenses and in biosyntheses and for synthesis of pentoses used as nucleic acid precursors. Because of the latter fact, thiamine utilization is increased in tumor cells. Thiamine uptake by the small intestines and by cells within various organs is mediated by a saturable, high affinity transport system. Alcohol affects thiamine uptake and other aspects of thiamine utilization, and these effects may contribute to the prevalence of thiamine deficiency in alcoholics. The major manifestations of thiamine deficiency in humans involve the cardiovascular (wet beriberi) and nervous (dry beriberi, or neuropathy and/or Wernicke-Korsakoff syndrome) systems. A number of inborn errors of metabolism have been described in which clinical improvements can be documented following administration of pharmacological doses of thiamine, such as thiamine-responsive megaloblastic anemia. Substantial efforts are being made to understand the genetic and biochemical determinants of inter-individual differences in susceptibility to development of thiamine deficiency-related disorders and of the differential vulnerabilities of tissues and cell types to thiamine deficiency. PMID- 11899072 TI - Macrophage response to Mycobacterium tuberculosis during HIV infection: relationships between macrophage activation and apoptosis. AB - Human macrophages represent the first line of defense for the containment of Mycobacterium tuberculosis infection. After phagocytosis, macrophages express activation surface markers and produce proinflammatory cytokines and chemokines whose main role is to control pathogen spreading by recruiting peripheral lymphocytes and monocytes at the site of inflammation. However, in the case of a concomitant human immunodeficiency virus (HIV) infection, these signals strongly enhance the susceptibility to viral infection both at the viral entry and replication levels. Under these conditions, viral expansion extends beyond tissue macrophages to T cells and vice-versa, according to the emerging viral phenotype. In absence of an efficient immune response, Mycobacterium tuberculosis can replicate in macrophages in an uncontrolled fashion culminating in macrophage death by apoptosis. As a consequence, a more severe form of immunedepression, involving both innate and specific immune responses, could be responsible for both ematogenous mycobacterial dissemination and extrapulmonary form of tuberculosis in HIV-infected patients. PMID- 11899073 TI - DNA vaccines. AB - Within the last decade bacterial plasmids encoding foreign antigens have revolutionized vaccine design. Although no DNA vaccine has yet been approved for routine human or veterinary use, the potential of this vaccine modality has been demonstrated in experimental animal models. Plasmid DNA vaccination has shown efficacy against viral, bacterial and parasitic infections, modulated the effects of autoimmune and allergic diseases and induced control over cancer progression. With a better understanding of the basic immune mechanisms that govern induction of protective or curative immune responses, plasmid DNA vaccines and their mode of delivery are continuously being optimized. Because of the simplicity and versatility of these vaccines, various routes and modes of delivery are possible to engage the desired immune responses. These may be T or B effector cell responses able to eliminate infectious agents or transformed cells. DNA vaccines may also induce an immunoregulatory/modulatory or immunosuppressive (tolerizing) response that interferes with the differentiation, expansion or effector functions of B and T cells. In this sense a DNA vaccine may be thought of as a 'negative' vaccine. Pre-clinical and initial small-scale clinical trials have shown DNA vaccines in either of these modes to be safe and well tolerated. Although DNA vaccines induce significant immune responses in small animal trials their efficacy in humans has so far been less promising thus necessitating additional optimizations of this novel vaccine approach. PMID- 11899074 TI - Adeno-associated virus (AAV) as a vehicle for therapeutic gene delivery: improvements in vector design and viral production enhance potential to prolong graft survival in pancreatic islet cell transplantation for the reversal of type 1 diabetes. AB - Most viral gene delivery syslems utilized to date have demonstrated significant limitations in practicality and safety due to the level and duration of recombinant transgene expression as well as their induction of host immunogenicity to vector proteins. Recombinant adeno-associated virus (rAAV) vectors appear to offer a vehicle for safe, long-term therapeutic gene transfer; factors afforded through the propensity of rAAV to establish long-term latency without deleterious effects on the host cell and the relative non-immunogenicity of the virus or viral expressed transgenes. The principal historical limitation of this vector system, efficiency of rAAV-mediated transduction, has recently observed a dramatic increase as the titer, purity, and production capacity of rAAV preparations have improved. In terms of systems that could benefit from such improvements, rAAV gene therapy to enhance solid organ transplantation would appear an obvious choice with islet transplantation forming a promising candidate due to the ability to perform viral transductions ex vivo. Currently, islet transplantation can be used to treat type 1 diabetes yet persisting alloimmune and autoimmune responses represent major obstacles to the clinical success for this procedure. The delivery of transgenes capable of interfering with antigenic recognition and/or cell death [e.g., Fas ligand (FasL), Bcl-2, Bcl-XL] as well as imparting tolerance/immunoregulation [e.g., interleukin(IL)-4, IL-10, transforming growth factor (TGF)-beta], or cytoprotection [e.g., heme oxygenase-1 (HO-1), catalase, manganese superoxide dismutase (MnSOD)] may prevent recurrent type 1 diabetes in islet transplantation and offer a promising form of immunotherapy. Research investigations utilizing such systems may also provide information vital to understanding the immunoregulatory mechanisms critical to the development of both alloimmune and autoimmune islet cell rejection mechanisms and recurrent type 1 diabetes. PMID- 11899075 TI - Molecular analysis of primary central nervous system and primary intraocular lymphomas. AB - Primary central nervous system lymphoma (PCNSL) is usually a large B-cell, high grade non-Hodgkin's lymphoma (NHL) classified as a diffuse large cell lymphoma (DLCL). In rare cases, however, T cell lymphomas have been described. Although a relatively rare tumor, the incidence of PCNSL has increased dramatically over the past 15 years in both immuno-competent and immunocompromised patients. The disease is aggressive with a 5-year survival rate of less than 25 %. The cause of death is progressive and recurrent disease in the CNS, despite aggressive treatment. Approximately 20-25% of patients with PCNSL also have primary intra ocular lymphoma (PIOL). PCNSL and PIOL are closely related and inter-connected pathologies involving two immune privileged sites. The study of PCNSL and PIOL has been limited due to the fact that viable malignant cells are rare and difficult to recognize. Moreover, the cells are difficult to culture and to date there is no good animal model for the disease. Here, we will present the current literature on the disease. In particular, we will present data suggesting that PCNSL in immuno-compromised and AIDS patients may correspond to two different pathologies. Furthermore, we will discuss how the study of these lymphomas can benefit from new advanced molecular biology techniques including single cell PCR and laser capture microdissection (LCM). PCNSL and PIOL are aggressive tumors, therefore, early diagnosis and prompt, aggressive treatment may improve prognosis. Advanced molecular biology will help delineate the oncogenesis of PCNSL and PIOL. PMID- 11899076 TI - Diabetes mellitus-cell transplantation and gene therapy approaches. AB - Diabetes mellitus affects millions of people in the United States and worldwide. It has become clear over the past decade that the chronic complications of diabetes result from lack of proper blood glucose concentration regulation, and particularly the toxic effects of chronic hyperglycemia on organs and tissues. Pancreas transplants can cure insulin-dependent diabetes mellitus (IDDM). Furthermore, recent advances in pancreatic islet isolation and immunosuppressive regimens have resulted in dramatic improvements in the survival and function of islet allografts. Therefore, islet replacement strategies are becoming increasingly attractive options for patients at risk for severe diabetic complications. A major limitation of these approaches is the small number of organs available for transplantation or islet isolation. Thus, an important next step in developing curative treatments for type I diabetes will be the generation of a replenishable source of glucose-responsive, insulin-secreting cells that can be used for beta cell replacement. This review focuses on approaches to developing robust and widely applicable beta-cell replacement strategies with an emphasis on manipulating beta-cell growth and differentiation by genetic engineering. PMID- 11899077 TI - Role of the NF-kappaB pathway in the pathogenesis of human disease states. AB - The NF-kappaB family consists of a group of inducible transcription factors which regulate immune and inflammatory responses and protect cells from undergoing apoptosis in response to cellular stress. A number of signal transduction cascades can activate the NF-kappaB pathway to result in the translocation of the NF-kappaB proteins from the cytoplasm to the nucleus where they activate the expression of specific cellular genes. In this review, we discuss cellular genes which are regulated by NF-kappaB and disease states which are associated with constitutive activation of the NF-kappaB pathway. Strategies to prevent prolonged activation of the NF-kappaB pathway are also discussed. PMID- 11899078 TI - Molecular characterization of the T cell repertoire using immunoscope analysis and its possible implementation in clinical practice. AB - T lymphocytes play a central role in the pathogenesis of a large number of human conditions including autoimmunity and graft rejection. Although T cells are key players in mounting immune responses, the assessment of T cell repertoires has yet to find an important role in clinical decision making. In this review, we discuss the "immunoscope" technique and its potential diagnostic role in a variety of clinical scenarios. This is an RT-PCR based approach that subdivides a bulk T cell population (i. e. from blood, lymph, spleen, or tissue) into approximately 2800 groups based upon rearranged variable beta (Vbeta)/joining beta (Jbeta) gene segments and the resulting length of the T cell receptor's (TCR's) third complementarity determining region (CDR-3). This extensive subdivision, or focusing, allows clonal expansions to be directly observed. Such a fine-tuned analysis has revealed previously unappreciated aspects of the T cell repertoire. For instance, an antigen-specific immune response can be divided into both public and non-public components. The non-public repertoire contains the majority of the expanding T cells which are unique to the individual (private), or shared by only some (semi-private), while "public" T cells can be found responding to the antigenic determinant in every individual. Although they are often a minority of the response, the public T cell repertoire seems to play a more important role in defining, as well as driving, the overall immune phenotype in the animal. Immunoscope analysis has identified public and non-public responses in human pathologies, such as multiple sclerosis. The ability to characterize the driver T cells dictating the state of immunity/autoimmunity in individual patients will be an important step towards understanding autoimmunity and designing effective treatment for a variety of conditions including rheumatoid arthritis and multiple sclerosis. We review the current literature involving public and non-public repertoires and discuss the prospect that immunoscope analysis may play a central role in the study and perhaps the management of human autoimmune diseases, and cancer. PMID- 11899079 TI - Alternative routes for the formation of immunochemically distinct advanced glycation end-products in vivo. AB - The advanced stage of the glycation process (also called the "Maillard reaction") that leads to the formation of advanced glycation end-products (AGEs) plays an important role in the pathogenesis of angiopathy in diabetic patients and in the aging process. AGEs elicit a wide range of cell-mediated responses that might contribute to diabetic complications, vascular disease, renal disease, and Alzheimer's disease. Recently, it has been proposed that AGE are not only created from glucose per se, but also from dicarbonyl compounds derived from glycation, sugar autoxidation, and sugar metabolism. However, this advanced stage of glycation is still only partially characterized and the structures of the different AGEs that are generated in vivo have not been completely determined. Because of their heterogeneity and the complexity of the chemical reactions involved, only some AGEs have been characterized in vivo, including N carboxymethyllysine (CML), pentosidine, pyrraline, and crosslines. In this article, we provide a brief overview of the pathways of AGE formation and of the immunochemical methods for detection of AGEs, and we also provide direct immunological evidence for the existence of five distinct AGE classes (designated as AGE-1 to -5) within the AGE-modified proteins and peptides in the serum of diabetic patients on hemodialysis. We also propose pathways for the in vivo formation of various AGEs by glycation, sugar autoxidation, and sugar metabolism. PMID- 11899080 TI - Molecular steps of tumor necrosis factor receptor-mediated apoptosis. AB - Until recently it was believed that TNF-induced apoptosis is mediated exclusively by TNF-RI because TNF-RII lacks death domain. However, it has been demonstrated that TNF-RII enhances TNF-RI-mediated apoptosis. In this review, I have discussed the evidence and mechanisms by which TNF-RII regulates TNF-alpha-induced apoptosis. A role of RIP is emphasized and novel mechanisms of FLIP-mediated inhibition of apoptosis are discussed. In addition, various mechanisms of TNF induced activation of mitochondrial pathway of apoptosis have been reviewed. PMID- 11899081 TI - Gene therapy for diabetes mellitus. AB - There are diverse strategies for gene therapy of diabetes mellitus. Prevention of beta-cell autoimmunity is a specific gene therapy for prevention of type 1 (insulin-dependent) diabetes in a preclinical stage, whereas improvement in insulin sensitivity of peripheral tissues is a specific gene therapy for type 2 (non-insulin-dependent) diabetes. Suppression of beta-cell apoptosis, recovery from insulin deficiency, and relief of diabetic complications are common therapeutic approaches to both types of diabetes. Several approaches to insulin replacement by gene therapy are currently employed: 1) stimulation of beta-cell growth, 2) induction of beta-cell differentiation and regeneration, 3) genetic engineering of non-beta cells to produce insulin, and 4) transplantation of engineered islets or beta cells. In type 1 diabetes, the therapeutic effect of beta-cell proliferation and regeneration is limited as long as the autoimmune destruction of beta cells continues. Therefore, the utilization of engineered non beta cells free from autoimmunity and islet transplantation with immunological barriers are considered potential therapies for type 1 diabetes. Proliferation of the patients' own beta cells and differentiation of the patients' own non-beta cells to beta cells are desirable strategies for gene therapy of type 2 diabetes because immunological problems can be circumvented. At present, however, these strategies are technically difficult, and transplantation of engineered beta cells or islets with immunological barriers is also a potential gene therapy for type 2 diabetes. PMID- 11899082 TI - Mechanisms of T cell receptor antagonism: implications in the treatment of disease. AB - The adaptive immune response is often required for the successful clearing of infectious pathogens. Antigen presenting cells (APC) present peptide antigens derived from pathogens to T cells via major histocompatibility complex (MHC) molecules. T cells then become activated and differentiate into effector cells with the capacity to kill infected cells or to induce an anti-pathogen antibody response. In autoimmunity, this T cell response is directed against self-antigens and often leads to deleterious effects on specific tissues. Likewise, T cell responses to allogeneic MHC molecules in transplants also leads to pathology. By introducing subtle changes in the antigenic peptide amino acid content, T cell activation can be inhibited, thereby preventing T cell effector functions. This strategy of TCR antagonism has been used successfully in vitro and in vivo to inhibit models of autoimmunity and allorecognition. In addition, a variety of pathogens that often result in chronic disease following infection, also have seemingly evolved natural mechanisms to inhibit T cell responses by antagonism. These microorganisms express natural variants of certain proteins, that when presented to T cells have the capacity to specifically inhibit T cell responses by functioning as antagonists or by modulating the nature of the T cell response. The understanding of how pathogens mediate this inhibition in vivo will be beneficial to ongoing studies in both autoimmunity and transplantation aimed at suppressing the harmful immune response, thereby controlling disease. TCR antagonism seems to have the potential to be used therapeutically to prevent or inhibit an undesired T cell response that will ultimately lead to disease. PMID- 11899083 TI - Insights into the pathogenesis of type 1 diabetes: a hint for novel immunospecific therapies. AB - Type 1 diabetes is an organ-specific autoimmune disease whose incidence is increasing worldwide. At present, there is no effective therapy to prevent or cure this disease. The genetic background (MHC and non-MHC genes) and environmental factors (pathogens, drugs, and diet) are critical for the initiation of the autoimmune response against the pancreatic beta-cells. Recognition of the pancreatic autoantigens by T cells in a predetermined environment of antigen-presenting cells, costimulation, and cytokines is crucial for the selective activation of diabetogenic or protective/regulatory T cells. Once the autoimmune process is triggered, epitope spreading and sustaining the autoimmune responses by continuous antigen stimulation leads to expansion of effector cells, which launch the attack on the beta-cells. Despite of some controversy, most of the studies in humans and animal models suggest that CD4 (Th1) T cells are directly involved in the autoimmune attack by secretion of pro inflammatory cytokines and recruitment of cytotoxic CD8 T cells. Secretion of anti-inflammatory cytokines by Th2 cells is protective against the disease. Therapy with peptides derived from major target antigens, such as glutamic acid decarboxylase 65 or proinsulin, can prevent the disease in animal models by rising protective Th2 cells. Herein, we review the recent progress in the immunopathogenesis of Type 1 diabetes and insights into the development of new diagnostic tools and antigen-specific immunomodulators, such as MHC-peptide chimeras. PMID- 11899084 TI - Pediatric autoimmune liver diseases: the molecular basis of humoral and cellular immunity. AB - Pediatric autoimmune liver disease is mainly represented by two similar liver disorders: autoimmune hepatitis (AIH) and autoimmune sclerosing cholangitis (ASC), both characterized by hypergammalobulinemia, interface hepatitis and the presence of a wide range of circulating autoantibodies. Although similar features are seen in AIH and inflammatory bowel disease, histological biliary changes are more common in ASC. In addition to their role as diagnostic markers, autoantibodies, such as anti-extractable nuclear antigen (ENA) antibodies and liver kidney microsomal antibody type 1 (LKM1) may be involved directly in inducing aggressive liver diseases. Although the cellular immune response in pediatric autoimmune liver disease has been less intensively investigated than humoral immunity, the importance of antigen specific T cells has been explored. Both alphabeta and gammadelta T cells derived from either peripheral blood and liver biopsies have highly heterogeneous TCR gene usage and cytolytic activity has been demonstrated. There have been attempts to seek triggers of liver autoimmunity and several sequences shared in common between autoantigens and hepatotropic viruses, namely hepatitis B, C and cytomegalovirus have been identified. The presence of cross-reactivity between homologous sequences, especially between HCV and cytochromes, supports the possibility that molecular mimicry plays a role in the induction of autoantibodies and autoreactive cytotoxic T cells. PMID- 11899085 TI - Directed gene modification via triple helix formation. AB - The ability to selectively target mammalian genes and disrupt or restore their function would represent an important advance in gene therapy. Mutation of a single nucleotide can often result in a non-functional gene product. Reversion of defective genes to their correct sequences could lead to permanent cures for patients with many genetic diseases. Molecules such as triplex forming oligonucleotides (TFOs) and peptide nucleic acids (PNAs) are currently being employed to bind to double-stranded DNA. Efficient targeting of genomic DNA with these molecules will be the initial step in gene modification. PMID- 11899086 TI - Histone acetylation/deacetylation and cancer: an "open" and "shut" case? AB - DNA in eukaryotic cells is packaged into chromatin. The main packaging component of chromatin is the nucleosome, and this is composed of proteins known as histones. Histones can be reversibly modified in several ways, and the best characterized of these modifications is histone acetylation. This is a reversible modification, which is carried out by two families of enzymes, the histone acetyltransferases (HATs), and the histone deacetylases (HDACs). These enzymes have important activities in many cellular processes including transcription, DNA replication and cell cycle progression. The mechanisms underlying tumor formation are multifaceted, and often involve mutations or alterations of genes involved with the regulation and control of the cell cycle or cell death. Because of their important roles in the regulation of such events, enzymes that affect histone acetylation status are increasingly being associated with tumors. This article describes some of the current knowledge about histone acetyltransferases and histone deacetylases, and how their multitudinal roles in cellular events may have important roles in tumorigensis. PMID- 11899087 TI - Cellular and molecular mechanisms of vaccine-induced protection against retroviral infections. AB - More than 15 years after the discovery of human immunodeficiency virus (HIV), researchers are still struggling to design a protective AIDS vaccine. A remaining problem is a lack of basic knowledge about the immunological requirements for protection against retroviruses. Infection of macaque monkeys with simian immunodeficiency virus is still the best model for HIV vaccine research. However, in this model it remains difficult to determine protective immunological mechanisms because of limited numbers of experimental animals and their genetic heterogeneity. Thus, fundamental concepts in retroviral immunology have to be defined in other ways such as mouse models. This minireview summarizes new findings on cellular and molecular mechanisms in protection of mice against Friend murine retrovirus infection. It has been shown that complex immune responses, including B and T cell responses, are required for efficient protection in this model. Multiple viral antigens are necessary to elicit such broad immune reactivity. Efficacious vaccines must protect not only against acute disease, but also against the establishment of persistent infections or the host is at serious risk of virus reactivation. The minireview closes with a discussion on the relevance of findings from the mouse model on the design of a protective vaccine against HIV. PMID- 11899088 TI - Biology of gammadelta T cells in tuberculosis and malaria. AB - Tuberculosis and malaria remain the leading causes of mortality among human infectious diseases in the world. It is estimated that 3 to 5 million people die from tuberculosis and malaria each year. Although it is traditionally believed that CD4 and CD8 alphabeta T lymphocytes are mandatory for protective immune responses against Mycobacterium tuberculosis and Plasmodium falciparum (the ethiologic agents of tuberculosis and the most severe form of malaria, respectively), there is still incomplete understanding of the mechanisms of immune protection and of the causes of its failure in the affected patients. Several studies in humans and animal models have suggested that Vgamma9/Vdelta2 T cells may play an important role in the immune responses against Mycobacterium tuberculosis and Plasmodium falciparum. Vgamma9/Vdelta2 T cells represent about 75% of all circulating gammadelta T cells while they can be greatly expanded during the acute phase of Mycobacterium tuberculosis and Plasmodium falciparum malaria. Vgamma9/Vdelta2 T recognize a new class of antigenic molecules which are nonpeptidic in nature and contain critical phosphate moieties (phosphoantigens). Interestingly, phosphoantigens isolated from Mycobacterium tuberculosis and Plasmodium falciparum share strong structural homology and are probably identical. However, despite a large body of data reported in the literature, it is not yet clear whether Vgamma9/Vdelta2 T cells play a protective or pathogenic role in immune responses against Mycobacterium tuberculosis and Plasmodium falciparum. In this review we summarize our current knowledge of the biology of Vgamma9/Vdelta2 T cells in response to the two pathogens, Mycobacterium tuberculosis and Plasmodium falciparum, and provide evidence suggesting definition of a novel and important protective role through which Vgamma9/Vdelta2 T cells can contribute to the killing of microorganisms residing in intracellular compartments. PMID- 11899089 TI - Restoring the phenotype of fragile X syndrome: insight from the mouse model. AB - A mouse model for the fragile X syndrome, the most common form of inherited mental retardation, was generated a number of years ago. It shows characteristics compatible with the clinical symptoms of human patients. These include pathological changes such as macroorchidism, behavioral problems, and diminished visuo-spatial abilities. To investigate whether the fragile X syndrome is a potentially correctable disorder, several groups attempted to 'rescue' the knockout mutation by introduction of an intact copy of the FMR1 gene in the knockout mouse. Two different types of rescue mice have been created by injection of constructs based on FMR1 cDNA or on FMR1 genomic DNA. Several pathological, behavioral and cognitive function tests were performed on these two different rescue mouse lines to compare their characteristics with those of the knockout and control littermates. Each rescue line resembled the control in some aspects though neither of the 2 lines was a full 'rescue', e.g. resemble the control in all aspects investigated. Thus, rescue of some aspects of the phenotype has been achieved by introduction of FMR1 constructs in the fragile X knockout mice. The results implicate that, even if FMR1 production is cell type specific, the quantity of the FMRP expression is highly critical as overproduction may have a harmful effect. PMID- 11899090 TI - Continuing education of the immune system--dendritic cells, immune regulation and tolerance. AB - T cells, as they develop in the thymus come to express antigen receptors. The specificity of these receptors cannot be predicted and must include many with potential anti-self reactivity. Those that encounter self-antigens, in association with self-MHC (major histocompatibility complex), with high affinity are inactivated and do not leave the thymus. Not all self-antigens however are expressed in the thymus and thus many potentially self-reactive T cells enter the periphery. It poses therefore a fundamental immunological question: how peripheral self-tolerance is maintained in health? Dendritic cells (DC) play a central role in the activation of T cells, especially naive T cells. Their importance in initiating immune responses against pathogens has been well established. However, DC represent complex populations of cells. Recent advances in our knowledge including molecular understanding of DC/T cell interactions have begun to reveal another important dimension of DC functions in the periphery, being not only initiators but also regulators of the immune system. This review summarises recent findings on the roles of DC in the regulation of immune responses and the maintenance of peripheral tolerance, in an attempt to explain how break down of this may lead to immunopathologies and autoimmunity. The concept of a regulatory DC and its possible role in the generation of T regulatory cells in health and in diseases are also discussed. Based on these, the need for a "continuing education" of the immune system throughout one's life, in which DC are again the "tutors", is postulated. PMID- 11899091 TI - Role of the mannose receptor in the immune response. AB - The mannose receptor (MR) recognizes a range of carbohydrates present on the surface and cell walls of micro-organisms. The MR is primarily expressed on macrophages and dendritic cells and is involved in MR-mediated endocytosis and phagocytosis. In addition, the MR plays a key role in host defense and provides a link between innate and adaptive immunity. Herein, we will review the role of the MR in innate host defense as well as the recent evidence for its role in the adaptive response, for both humoral and cellular immune responses. PMID- 11899092 TI - Genetic bases and medical relevance of capsular polysaccharide biosynthesis in pathogenic streptococci. AB - Many streptococci are human and/or animal pathogens and the frequent cause of life-threatening diseases. Among various streptococcal virulence factors, capsular polysaccharides (CPs) are recognized as essential to prevent phagocytosis by macrophages and neutrophils. In the last decade, an impressive advance on the knowledge of the genetic bases underlying capsule formation has been achieved. The capsular gene cluster driving the formation of the CP of Streptococcus pyogenes and other hyaluronate-producing streptococci, represents one of the simplest cases of gene organization to synthesize a capsule. A more complex situation has been found in Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus suis, and other streptococci. On the whole, there exists a direct relationship between the structural and chemical complexity of the repeating unit of the polysaccharide and the number of genes found in the corresponding capsular locus. Streptococcal vaccines, either polysaccharide or conjugate, are currently being tested in clinical trials to overcome the rise of worldwide antibiotic resistance, although, for different reasons, none of these vaccines are expected to provide the required full coverage in a near future. This concern has prompted to explore alternative possibilities with an improved therapeutic potential against streptococcal diseases. PMID- 11899094 TI - Gastric toxicity and mucosal ulceration induced by oxygen-derived reactive species: protection by melatonin. AB - Uncontrolled hydrochloric acid secretion and ulceration of the stomach mucosa due to various factors are serious global problems. Although the mechanism of acid secretion from the parietal cell is now well understood, the processes involved in gastric ulceration are still not clear. Among various causes of gastric ulceration, lesions caused by stress, alcohol consumption, Helicobacter pylori infection and due to use of nonsteroidal antiinflammatory drugs have been shown to be mediated largely through the generation of reactive oxygen species, especially the hydroxyl radical. A number of excellent drugs have proven useful in controlling hyperacidity and ulceration but their long-term use is associated with disturbing side-effects. Hence, the search is still on to find a compound possessing antisecretory, antiulcer and antioxidant properties which will serve as a therapeutic agent to reduce gastric hyperacidity and ulcers. This article describes the role of reactive oxygen species in gastric ulceration, drugs controlling them with their merits and demerits and, the role of melatonin, a pineal secretory product, in protecting against gastric lesions. In experimental studies, melatonin has been shown to be effective in reducing mucosal breakdown and ulcer formation in a wide variety of situations. Additionally, the low toxicity of melatonin supports further investigation of this molecule as a gastroprotective agent. Finally, we include a commentary on how melatonin research with respect to gastric pathophysiology can move forward with a view of eventually using this indole as a therapeutic agent to control gastric ulceration in humans. PMID- 11899095 TI - Making the animal model for AIDS research more precise: the impact of major histocompatibility complex (MHC) genes on pathogenesis and disease progression in SIV-infected monkeys. AB - Experimentally infected rhesus monkeys serve as an indispensable animal model to assess the pathogenesis, to validate therapy approaches and to develop vaccination strategies against viral diseases such as AIDS threatening the human population. Upon infection with simian immunodeficiency virus (SIV), a retrovirus closely related to the human immunodeficiency virus (HIV), macaques develop clinical manifestations similar to those of HIV-infected humans. As in humans, the disease course is variable. Polymorphic genes of the major histocompatibility complex (MHC) are required for the initiation and regulation of a specific immune response and represent a major host factor accounting for the differential outcome of infection. During the last few years, our understanding of the structure and function of the rhesus macaque MHC has increased substantially. Functional studies have led to the identification of specific SIV and HIV peptide epitopes presented by rhesus macaque MHC molecules. The subsequent development of MHC class I tetramers has allowed further insight into the cellular immune response following SIV-infection. Detailed studies demonstrated that viral escape mutants are generated during the acute and chronic phase of infection and explain why control of viral replication ultimately fails. Furthermore, particular MHC haplotypes which influence disease progression have been discovered. Thus, MHC typing can have a prognostic potential. The further elucidation of the rhesus macaque MHC and the search for other relevant genes will remain an important task for future research and will stimulate all immunologically-related investigations in macaques. PMID- 11899093 TI - Gene therapy: a battle against biological barriers. AB - One factor critical to successful human gene therapy is development of efficient gene delivery systems. Although numerous vector systems for gene transfer have been developed, a perfect vector system has not yet been constructed. Difficulties of in vivo gene transfer appear to result from resistance of living cells to invasion by foreign materials and from interference of cellular functions. We should analyze what barriers in tissues affect in vivo gene transfection and how to solve these problems for gene therapy. In this review article, the biological barriers to in vivo gene transfection are discussed and possible solutions to each barrier are discussed with respect to construction of a perfect gene therapy vector system. PMID- 11899096 TI - Melatonin as a chronobiotic/anticancer agent: cellular, biochemical, and molecular mechanisms of action and their implications for circadian-based cancer therapy. AB - Melatonin, as a new member of an expanding group of regulatory factors that control cell proliferation and loss, is the only known chronobiotic, hormonal regulator of neoplastic cell growth. At physiological circulating concentrations, this indoleamine is cytostatic and inhibits cancer cell proliferation in vitro via specific cell cycle effects. At pharmacological concentrations, melatonin exhibits cytotoxic activity in cancer cells. At both physiological and pharmacological concentrations, melatonin acts as a differentiating agent in some cancer cells and lowers their invasive and metastatic status through alterations in adhesion molecules and maintenance of gap junctional intercellular communication. In other cancer cell types, melatonin, either alone or in combination with other agents, induces apoptotic cell death. Biochemical and molecular mechanisms of melatonin's oncostatic action may include regulation of estrogen receptor expression and transactivation, calcium/calmodulin activity, protein kinase C activity, cytoskeletal architecture and function, intracellular redox status, melatonin receptor-mediated signal transduction cascades, and fatty acid transport and metabolism. A major mechanism mediating melatonin's circadian stage-dependent tumor growth inhibitory action is the suppression of epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) activity. This occurs via melatonin receptor-mediated blockade of tumor linoleic acid uptake and its conversion to 13-hydroxyoctadecadienoic acid (13-HODE) which normally activates EGFR/MAPK mitogenic signaling. This represents a potentially unifying model for the chronobiological inhibitory regulation of cancer growth by melatonin in the maintenance of the host/cancer balance. It also provides the first biological explanation of melatonin-induced enhancement of the efficacy and reduced toxicity of chemo- and radiotherapy in cancer patients. PMID- 11899097 TI - Melatonin, mitochondrial homeostasis and mitochondrial-related diseases. AB - The recently described 'hydrogen hypothesis' invokes metabolic symbiosis as the driving force for a symbiotic association between an anaerobic, strictly hydrogen dependent organism (the host) and an eubacterium (the symbiont) that is able to respire, but which generates molecular hydrogen as an end product of anaerobic metabolism. The resulting proto-eukaryotic cell would have acquired the essentials of eukaryotic energy metabolism, evolving not only aerobic respiration, but also the cost of oxygen consumption, i.e., generation of reactive oxygen species (ROS) and oxidative damage. Mitochondria contain their own genome with a modified genetic code that is highly conserved among mammals. Control of gene expression suggests that transcription of certain mitochondrial genes may be regulated in response to the redox potential of the mitochondrial membrane. Mitochondria are involved in energy production and conservation, and they have an uncoupling mechanism to produce heat instead of ATP. Also, mitochondria are involved in programmed cell death. Increasing evidence suggests the participation of mitochondria in neurodegenerative and neuromuscular diseases involving alterations in both nuclear (nDNA) and mitochondrial (mtDNA) DNA. Melatonin is now known as a powerful antioxidant and increasing experimental evidence shows its beneficial effects against oxidative stress-induced macromolecular damage and diseases, including those in which mitochondrial function is affected. This review summarizes the data and mechanisms of action of melatonin in relation to mitochondrial pathologies. PMID- 11899098 TI - Pharmacological actions of melatonin in acute and chronic inflammation. AB - A vast number of experimental and clinical studies implicates oxygen-derived free radicals (especially, superoxide and the hydroxyl radical) and high energy oxidants (such as peroxynitrite) as mediators of acute and chronic inflammation. The purpose of this review is to summarize the pharmacological actions of melatonin in acute and chronic inflammation. Reactive oxygen species can modulate a wide range of toxic oxidative reactions. These include initiation of lipid peroxidation, direct inhibition of mitochondrial respiratory chain enzymes, inactivation of glyceraldehyde-3-phosphate dehydrogenase, inhibition of membrane sodium/potassium ATPase activity, inactivation of membrane sodium channels, and other oxidative modifications of proteins. Reactive oxygen species (e.g., superoxide, peroxynitrite, hydrogen peroxide and hydroxyl radical) are all potential reactants capable of initiating DNA single strand breakage, with subsequent activation of the nuclear enzyme poly (ADP ribose) synthetase (PARS), leading to eventual severe energy depletion of the cells, and necrotic-type cell death. These toxic reactions are likely to play a role in the pathophysiology of inflammation. Melatonin has been shown to possess both in vitro and in vivo important antioxidant activities as well as to inhibit the activation of poly (ADP ribose) synthetase. A large number of experimental studies have documented that melatonin exerts important anti-inflammatory actions. PMID- 11899099 TI - Melatonin-immune system relationships. AB - In this paper we review the historical milestones that first highlighted the existence of a relationship between melatonin and the immune system and we summarize data from experiments which correlate the rhythmic production of melatonin with the rhythmic activity of the immune system. The effects of pinealectomy and in vivo administration of melatonin on a variety of immune parameters, including specific and non-specific immunity are considered and we also present contradictory data concerning the effect of melatonin in cultured immunocompetent cells and a possible scheme of how melatonin regulates the production of a number of cytokines. Finally, the mechanism of action of melatonin in the immune system is discussed. Many data suggest the existence of both nuclear and membrane receptors for melatonin in the immune system. Both of these appear to be clearly identified but their specific physiological role is still under discussion. In summary, although there is overwhelming information demonstrating the immunoenhancing properties of melatonin, many questions related to the cytokines involved and the mechanisms of action of the indoleamine require answers. PMID- 11899100 TI - Chemical and physical properties and potential mechanisms: melatonin as a broad spectrum antioxidant and free radical scavenger. AB - Melatonin was found to be a potent free radical scavenger in 1993. Since then over 800 publications have directly or indirectly confirmed this observation. Melatonin scavenges a variety of reactive oxygen and nitrogen species including hydroxyl radical, hydrogen peroxide, singlet oxygen, nitric oxide and peroxynitrite anion. Based on the analyses of structure-activity relationships, the indole moiety of the melatonin molecule is the reactive center of interaction with oxidants due to its high resonance stability and very low activation energy barrier towards the free radical reactions. However, the methoxy and amide side chains also contribute significantly to melatonin's antioxidant capacity. The N C=O structure in the C3 amide side chain is the functional group. The carbonyl group in the structure of N-C=O is key for melatonin to scavenge the second reactive species and the nitrogen in the N-C=O structure is necessary for melatonin to form the new five membered ring after melatonin's interaction with a reactive species. The methoxy group in C5 appears to keep melatonin from exhibiting prooxidative activity. If the methoxy group is replaced by a hydroxyl group, under some in vitro conditions, the antioxidant capacity of this molecule may be enhanced. However, the cost of this change are decreased lipophility and increased prooxidative potential. Therefore, in in vivo studies the antioxidant efficacy of melatonin appears to be superior to its hydroxylated counterpart. The mechanisms of melatonin's interaction with reactive species probably involves donation of an electron to form the melatoninyl cation radical or through an radical addition at the site C3. Other possibilities include hydrogen donation from the nitrogen atom or substitution at position C2, C4 and C7 and nitrosation. Melatonin also has the ability to repair damaged biomolecules as shown by the fact that it converts the guanosine radical to guanosine by electron transfer. Unlike the classical antioxidants, melatonin is devoid of prooxidative activity and all known intermediates generated by the interaction of melatonin with reactive species are also free radical scavengers. This phenomenon is defined as the free radical scavenging cascade reaction of the melatonin family. Due to this cascade, one melatonin molecule has the potential to scavenge up to 4 or more reactive species. This makes melatonin very effective as an antioxidant. Under in vivo conditions, melatonin is often several times more potent than vitamin C and E in protecting tissues from oxidative injury when compared at an equivalent dosage (micromol/kg). Future research in the field of melatonin as a free radical scavenger might be focused on: 1), signal transduction and antioxidant enzyme gene expression induced by melatonin and its metabolites, 2), melatonin levels in tissues and in cells, 3), melatonin structure modifications, 4), melatonin and its metabolites in plants and, 5), clinical trials using melatonin to treat free radical related diseases such as Alzheimer's, Parkinson's, stroke and heart disease. PMID- 11899101 TI - Melatonin and circadian rhythms. AB - Melatonin synthesis and secretion by the pineal gland is under the control of the suprachiasmatic nucleus and consequently has a profound circadian rhythm. In this review we discuss some of the issues surrounding the measurement of melatonin rhythmicity in biological fluids and the factors that influence melatonin circadian rhythmicity, including light and drugs. We also review the role of melatonin rhythmicity in sleep timing and sleep initiation. PMID- 11899102 TI - The new pre-preclinical paradigm: compound optimization in early and late phase drug discovery. AB - The attrition rates of new chemical entities (NCEs) in preclinical and clinical development are staggeringly high. NCEs are abandoned due to insufficient efficacy, safety issues, and economic reasons. Uncovering drug defects that produce these failures as early as possible in drug discovery would be highly effective in lowing the cost and time of developing therapeutically useful drugs. Unfortunately, there is no single factor that can account for these NCE failures in preclinical and clinical development since factors, such as solubility, pKa, absorption, metabolism, formulation, pharmacokinetics, toxicity and efficacy, to name a few, are all interrelated. In addition, there are many problems in scaling up drug candidates from the laboratory bench scale to the pilot plant scale. To address the problem of attrition rates of NCEs in preclinical and clinical development and drug scale-up issues, pharmaceutical companies need to reorganize their preclinical departments from a traditional linear approach to a parallel approach. In this review, a strategy is put forth to integrate certain aspects of drug metabolism/pharmacokinetics, toxicology functions and process chemistry into drug discovery. Compound optimization in early and late phase drug discovery occurs by relating factors such as physicochemical properties, in vitro absorption, in vitro metabolism, in vivo pharmacokinetics and drug scale-up issues to efficacy optimization. This pre-preclinical paradigm will improve the success rate of drug candidates entering development. PMID- 11899103 TI - Strategies for absorption screening in drug discovery and development. AB - This review gives an overview of the current approaches to evaluate drug absorption potential in the different phases of drug discovery and development. Methods discussed include in silico models, artificial membranes as absorption models, in vitro models such as the Ussing chamber and Caco-2 monolayers, in situ rat intestinal perfusion and in vivo absorption studies. In silico models such as iDEA can help optimizing chemical synthesis since the fraction absorbed (Fa) can be predicted based on structural characteristics only. A more accurate prediction of Fa can be obtained by feeding the iDEA model with Caco-2 permeability data and solubility data at various pH's. Permeability experiments with artificial membranes such as the filter-IAM technology are high-throughput and offer the possibility to group compounds according to a low and a high permeability. Highly permeable compounds, however, need to be further evaluated in Caco-2 cells, since artificial membranes lack active transport systems and efflux mechanisms such as P-glycoprotein (PgP). Caco-2 and other "intestinal-like" cell lines (MDCK, TC-7, HT29-MTX, 2/4/A1) permit to perform mechanistic studies and identify drug-drug interactions at the level of PgP. The everted sac and Ussing chamber techniques are more advanced models in the sense that they can provide additional information with respect to intestinal metabolism. In situ rat intestinal perfusion is a reliable technique to investigate drug absorption potential in combination with intestinal metabolism, however, it is time consuming, and therefore not suited for screening purposes. Finally, in vivo absorption in animals can be estimated from bioavailability studies (ratio of the plasma AUC after oral and i.v. administration). The role of the liver in affecting bioavailability can be evaluated by portal vein sampling experiments in dogs. PMID- 11899104 TI - Assessing the absorption of new pharmaceuticals. AB - The advent of more efficient methods to synthesize and screen new chemical compounds is increasing the number of chemical leads identified in the drug discovery phase. Compounds with good biological activity may fail to become drugs due to insufficient oral absorption. Selection of drug development candidates with adequate absorption characteristics should increase the probability of success in the development phase. To assess the absorption potential of new chemical entities numerous in vitro and in vivo model systems have been used. Many laboratories rely on cell culture models of intestinal permeability such as, Caco-2, HT-29 and MDCK. To attempt to increase the throughput of permeability measurements, several physicochemical methods such as, immobilized artificial membrane (IAM) columns and parallel artificial membrane permeation assay (PAMPA) have been used. More recently, much attention has been given to the development of computational methods to predict drug absorption. However, it is clear that no single method will sufficient for studying drug absorption, but most likely a combination of systems will be needed. Higher throughput, less reliable methods could be used to discover 'loser' compounds, whereas lower throughput, more accurate methods could be used to optimize the absorption properties of lead compounds. Finally, accurate methods are needed to understand absorption mechanisms (efflux-limited absorption, carrier-mediated, intestinal metabolism) that may limit intestinal drug absorption. This information could be extremely valuable to medicinal chemists in the selection of favorable chemo-types. This review describes different techniques used for evaluating drug absorption and indicates their advantages and disadvantages. PMID- 11899106 TI - Nuclear magnetic resonance as a tool in drug discovery, metabolism and disposition. AB - Nuclear magnetic resonance techniques have become critically important in the design of new pharmaceuticals, the characterization of drug-receptor interactions and metabolite identification. Advances in solvent suppression, coherent and incoherent magnetization transfer pathway selection, isotope editing and filtering, and diffusion filtering have made it possible to examine the interactions between small molecules and proteins or nucleic acids in great detail. Multiple schemes for high-throughput lead compound identification, metabolite screening and drug disposition have been proposed and reduced to practice. In particular, the coupling of NMR with other analytical methods, especially HPLC, combine the structural and dynamic detail available from NMR methods with the resolution and sensitivity of other analytical techniques. PMID- 11899105 TI - Metabolism profiling, and cytochrome P450 inhibition & induction in drug discovery. AB - To reduce the high attrition rates of NCEs in preclinical and clinical development uncovering pharmacokinetics, toxicokinetics, drug metabolism, and drug-drug interactions early in drug discovery would be highly valuable. There have been many in vitro screens developed for these areas that have higher sample throughput, which is consistent with the iterative cycle of a typical drug discovery research project. We have presented the present status and given detailed descriptions of biotransformation, metabolic stability assays, identification of drug metabolizing P450 enzymes, prediction of pharmacokinetic parameters from in vitro metabolism data, structure elucidation of metabolites, CYP450 inhibition assays and CYP450 induction assays from a drug discovery perspective. Strategies for the proper sequencing of primary and secondary assays employedfor drug metabolism and CYP450 inhibition & induction is discussed. PMID- 11899107 TI - Tailoring bioanalysis for PK studies supporting drug discovery. AB - Over the last years, there has been an exponentially growing need and interest to bring pharmacokinetic expertise into discovery. In order to allow a multidisciplinary selection and a higher attrition rate, both the in vivo and in vitro pharmacokinetic parameters of an ever increasing number of tentative new chemical entities are evaluated in an earlier phase of Drug Discovery. A higher attrition rate at the beginning of the pipeline should result in a lower attrition rate at a later stage in development. In this process, the bioanalytical laboratory has become increasingly important. Analytical strategies needed to be adapted to cope with novel experimental designs such as cassette dosing, cassette analysis or 96-well techniques. At the same time, HT-synthesis programs surfaced a broader variety of chemical classes to be investigated, disfavoring further generalization of analytical approaches. Progress in lab automation, improved chromatographic techniques and the proliferation of LC-MS/MS enabled the analyst to deal with these challenges much faster and with a higher level of confidence. Quality standards regarding method development and method validation, setting the boundaries for more than a decade, needed to be titrated to reach an optimal balance between speed and quality. This review will give an illustrative overview of the bioanalytical techniques and strategies used to support Drug Discovery, together with some pitfalls related to the overzealous use of new techniques. PMID- 11899108 TI - The use of on-line and off-line chromatographic extraction techniques coupled with mass spectrometry for support of in vivo and in vitro assays in drug discovery. AB - We have investigated various sample chromatographic extraction and sample preparation methods for liquid chromatography mass spectrometry analysis in order to increase the throughput of various in vivo and in vitro assays in support of drug discovery. The results indicated that direct plasma injection, although certainly faster than conventional protein precipitation for sample preparation, had problems associated with column longevity and overall robustness. Frequently a single study could not be completed without column replacement. On-line solid phase extraction, on the other hand, compared well with off-line solid phase extraction, using our LC extraction column design, as contamination of the extraction column was minimized by back flushing using the Gilson syringe pump. Finally, on-line solid phase extraction for support of Caco-2 permeability studies worked very well for both single components and mixtures as the matrix was much simpler, presenting fewer contamination problems. PMID- 11899109 TI - Assessing the potential toxicity of new pharmaceuticals. AB - Optimizing chemical structures to create potentially safe drugs during discovery and early development relies on a combination of predictive algorithms, screening, formal toxicology studies, and early clinical trials. Early in the process three critical questions emerge that must be answered by a detailed "profiling" approach. These questions are: 1) is there a correlation between the chemical structure and potential toxicity that can be used to optimize structures of lead compounds, 2) can specific markers of potential toxicity can be identified carly and used as mechanistic decision-making screens, and 3) will exposures (plasma levels) in animal studies correlate with exposures encountered in the clinic thereby providing "coverage" for safety? Depending on the therapeutic class of compounds being considered and the level of knowledge available, feedback loops of information can be established to guide the development process. PMID- 11899110 TI - Integration of computational analysis as a sentinel tool in toxicological assessments. AB - Computational toxicity modeling can have significant impact in the drug discovery process, especially when utilized as a sentinel filter for common drug safety liabilities, such as mutagenicity, carcinogenicity and teratogenicity. This review will focus on the strengths and limitations of the current computational models for predicting these drug safety liabilities, and the various strategies for incorporating these predictive models into the drug discovery process. PMID- 11899111 TI - "Holistic" in silico methods to estimate the systemic and CNS bioavailabilities of potential chemotherapeutic agents. AB - A fundamental fact in the drug development process is that physical quantity of chemical substance is needed for experimental determinations. Information that could guide the course of a drug discovery program, in particular the penultimate ADME parameters % oral bioavailability (%F) and CNS permeability (BBB) are not explicitly determined until after large capital, human and time resources have been invested in a particular chemical series to produce the substance. To assure better go/no-go decisions and to protect the risks of a process that necessitates a considerable "front-loading" of resources, project teams are turning to computational methods to estimate these parameters. Herein we provide a detailed review of "holistic" in silico methods toward the estimation of %F and BBB. An unbiased description of the scope and limitations of their installation and application will be given in the context of an on going pharmaceutical project. PMID- 11899112 TI - Physicochemical profiling (solubility, permeability and charge state). AB - About 30% of drug candidate molecules are rejected due to pharmacokinetic-related failures. When poor pharmaceutical properties are discovered in development, the costs of bringing a potent but poorly absorbable molecule to a product stage by "formulation" can become very high. Fast and reliable in vitro prediction strategies are needed to filter out problematic molecules at the earliest stages of discovery. This review will consider recent developments in physicochemical profiling used to identify candidate molecules with physical properties related to good oral absorption. Poor solubility and poor permeability account for many PK failures. FDA's Biopharmaceutics Classification System (BCS) is an attempt to rationalize the critical components related to oral absorption. The core idea in the BCS is an in vitro transport model, centrally embracing permeability and solubility, with qualifications related to pH and dissolution. The objective of the BCS is to predict in vivo performance of drug products from in vitro measurements of permeability and solubility. In principle, the framework of the BCS could serve the interests of the earliest stages of discovery research. The BCS can be rationalized by considering Fick's first law, applied to membranes. When molecules are introduced on one side of a lipid membrane barrier (e.g., epithelial cell wall) and no such molecules are on the other side, passive diffusion will drive the molecules across the membrane. When certain simplifying assumptions are made, the flux equation in Fick's law reduces simply to a product of permeability and solubility. Many other measurable properties are closely related to permeability and solubility. Permeability (Pe) is a kinetic parameter related to lipophilicity (as indicated by the partition and distribution coefficients, log P and log D). Retention (R) of lipophilic molecules by the membrane (which is related to lipophilicity and may predict PK volumes of distribution) influences the characterization of permeability. Furthermore, strong drug interactions with serum proteins can influence permeability. The unstirred water layer on both sides of the membrane barrier can impose limits on permeability. Solubility (S) is a thermodynamic parameter, and is closely related to dissolution, a kinetic parameter. The unstirred water layer on the surfaces of suspended solids imposes limits on dissolution. Bile acids effect both solubility and dissolution, by a micellization effect. For ionizable molecules, pH plays a crucial role. The charge state that a molecule exhibits at a particular pH is characterized by the ionization constant (pKa) of the molecule. Buffers effect pH gradients in the unstirred water layers, which can dramatically affect both permeability and dissolution of ionizable molecules. In this review, we will focus on the emerging instrumental methods for the measurement of the physicochemical parameters Pe, S, pKa, R, log P, and log D (and their pH profiles). These physicochemical profiles can be valuable tools for the medicinal chemists, aiding in the prediction of in vivo oral absorption. PMID- 11899113 TI - Prostate cancer screening practices and cancer control research (United States). PMID- 11899114 TI - Birth characteristics, maternal reproductive history, hormone use during pregnancy, and risk of childhood acute lymphocytic leukemia by immunophenotype (United States). AB - OBJECTIVE: To investigate the associations of birth characteristics and maternal reproductive factors with risk of childhood acute lymphoblastic leukemia (ALL) by immunophenotypic subtypes. METHODS: Data collected from a case-control study including 1842 ALL cases (age < 15 years) and 1986 individually matched controls were analyzed. Exposure information was obtained through telephone interviews of parents. RESULTS: Factors associated with risk of ALL from all subgroups combined included high birth weight (OR = 1.4, 95% CI = 1.1-1.8), high birth order (OR = 2.0, 95% CI = 1.3-3.0 for fourth-born child compared to first-born child). young maternal age (<20 compared to 25-29, OR = 1.4, 95% CI = 1.1-1.9), advanced paternal age (>39 compared to 25-29, OR = 1.4, 95% CI = 1.0-1.9), induced abortion prior to the index pregnancy (OR = 1.2, 95% CI = 1.0-1.4), and oral contraceptive use during the index pregnancy (OR = 1.5, 95% CI = 1.0-2.2) with children under the age of 2 (OR = 5.1, 95% CI = 1.0-24.7) being the predominantly affected group. Risk of early pre-B-cell ALL increased with advanced paternal age (OR = 1.7, 95% CI = 1.1-2.7) and high birth order (OR = 2.0, 95% CI = 1.1-3.6), while risk of pre-B-cell ALL increased with both younger (OR = 3.4, 95% CI = 1.4 8.4) and advanced maternal age (OR = 2.6, 95% CI = 1.1-5.9). T-cell ALL was associated with high birth weight (OR = 2.4, 95% CI = 1.1-5.5) and history of induced abortion (OR = 2.4, 95% CI = 1.3-4.5). CONCLUSION: This study suggests that the association of ALL with birth characteristics and maternal reproductive factors varies with the immunophenotype of the ALL. Future studies are needed to better understand the effect of maternal hormone in the development of subtype of childhood ALL. PMID- 11899115 TI - The importance of smoking and medical history for development of small bowel carcinoid tumor: a European population-based case-control study. AB - OBJECTIVE: Little is known about the etiology of small bowel carcinoid tumor (SBC), but a few studies have pointed to certain medical and lifestyle factors as potential risk factors. This study aims to evaluate these findings and to identify new associations. METHODS: A population-based European multicenter case control study was conducted from 1995 through 1997. Incident histologically verified 35-69 year-old SBC cases (n = 99) and 3335 controls were recruited; 84 cases and 2070 controls were interviewed. RESULTS: Ever being a smoker was associated with SBC (odds ratio = 1.9; 95% confidence interval 1.1-3.2) and increased risk estimates were seen for all smoking categories. SBC was associated with previous gallstone disease and ovariectomy, but only when these conditions occurred within two years prior to the SBC diagnosis. No association was seen for a history of cholecystitis, liver cirrhosis, ulcerative disease, or Crohn's disease. Intake of alcoholic beverages--as well as medical treatments with radioactive substances, hormones, or corticosteroid tablets--were not associated with SBC. CONCLUSIONS: This study indicates that tobacco smoking is a risk factor for SBC. The associations with gallstone and ovarian diseases may be due to enhanced medical surveillance during the early phase of the cancer disease. PMID- 11899116 TI - Risk of epithelial ovarian cancer in relation to use of antidepressants, benzodiazepines, and other centrally acting medications. AB - OBJECTIVE: An increased risk of ovarian cancer among users of antidepressants and benzodiazepines has been observed in some but not all prior studies. We examined these associations in a population-based case-control study. METHODS: We identified 314 members of a health maintenance organization (HMO) who were diagnosed with epithelial ovarian cancer between 1981 and 1997, were aged 35-79 years at diagnosis, and had at least 4 years of HMO membership. Up to four controls were selected for each case (n = 790), matched on age, calendar year, and length of HMO membership. Information concerning past medication use was obtained from the computerized pharmacy database, established in 1977. RESULTS: Cases were slightly less likely than controls to have filled two antidepressant prescriptions (primarily for doxepin, amitriptyline, or imipramine) in any 6 month period prior to a reference date set 1.5 years before diagnosis (conditional odds ratio (OR) 0.71, 95% confidence interval (CI) 0.47-1.1), or to have used an antidepressant continuously for 6 months or longer (OR 0.64, 95% CI 0.36-1.1). Cases were less likely than controls to have filled two benzodiazepine prescriptions in 6 months (OR 0.70, 95% CI 0.47-1.0) or to have used benzodiazepines continuously for 6 months or longer (OR 0.53, 95% Cl 0.15-1.9). CONCLUSIONS: Our findings suggest that there is not an increased risk of ovarian cancer in women who have taken some types of antidepressants or benzodiazepines. PMID- 11899117 TI - Childbearing and the risk of leukemia in Sweden. AB - OBJECTIVE: The possible influence of childbearing on the development of leukemias in females has received little attention, in spite of consistent findings of lower incidence rates in females than in males. A nested case-control study was undertaken to explore if parity and age at first birth affect the risk of developing these malignancies. METHODS: In a nationwide cohort defined by a population-based Fertility Register, we identified 356 women with chronic myeloid leukemia (CML), 819 with acute nonlymphocytic leukemia (ANLL), and 179 with acute lymphocytic leukemia (ALL). For each case, five age-matched controls were selected. Odds ratios were estimated by conditional logistic regression analyses. RESULTS: There was some evidence of weak negative associations between parity and age at first birth for CML. Compared to nulliparous women there was a tendency of a temporal risk reduction of CML for the first 10 years following a delivery. The risk of ANLL was slightly lower in parous compared to nulliparous women. Neither parity nor age at first birth was related to the risk of ALL. CONCLUSIONS: We conclude that if pregnancy-related hormonal or immunological factors have an effect on the development of leukemia, it is minor and confined to the myeloid types, chiefly CML. Our study gives some support for treating the leukemias as separate entities based on both cell lineage and form in future etiologic studies. PMID- 11899119 TI - Stage-specific incidence of breast cancer before the beginning of organized screening programs in Italy. AB - OBJECTIVE: To measure stage-specific geographic and time variability of breast cancer in seven Italian areas before the onset of organized screening programs. METHODS: All invasive cancers (8689 cases) arising in women aged 40-79 years during the pre-screening period 1985-1997, were considered. Multiple Poisson regression analysis was performed. RESULTS: About 39% of the cases were classified as "early," 52% as "advanced," and 9% as "unspecified" stage. Age adjusted incidence rates showed a significant geographic variation for early but not for advanced cancers (range: 58-103 cases/100,000 and 104-125 cases/100,000, respectively). The result was confirmed in the multiple regression analysis after adjustment for year of diagnosis and age. Early breast cancer risk adjusted for age and registry showed a significant increase over time (+3.9% per year for all ages, and +6.2% per year for age category 50-79). In contrast, a decreasing time trend was observed for advanced cancer of 3 cm or over in women aged less than 60. CONCLUSIONS: In our study, early breast cancer incidence varied both by geographic area and time before the commencement of screening. The differences in early-stage incidence may well be related to differences in availability of "spontaneous" mammography. Late-stage incidence decreased over time in younger women and for very advanced cases, but not in the older ones, nor for cancers less than 3 cm. Early detection outside organized screening was only partially efficient in reducing advanced breast cancer incidence. The trend of incidence of advanced disease, as previously proposed, is confirmed to be a valid early indicator of effectiveness of screening. PMID- 11899118 TI - Body mass index, tobacco chewing, alcohol drinking and the risk of oral submucous fibrosis in Kerala, India. AB - OBJECTIVE: While chewing areca nut is considered a risk factor for oral submucous fibrosis, the effects of cigarette smoking, alcohol drinking, and body mass index (BMI) have not been examined; nor are they well established. In this study we investigated the association between BMI, smoking, drinking, and the risk of oral submucous fibrosis. METHODS: We conducted a case-control study within the framework of an ongoing randomized oral cancer screening trial in Kerala, India. Trained health workers conducted interviews with structured questionnaires and oral visual inspections to diagnose oral premalignant lesions. A total of 170 oral submucous fibrosis cases (139 women and 31 men) and 47,773 controls were identified. The odds ratios (OR) and 95% confidence intervals (CI) were calculated by logistic regression in SAS. RESULTS: The adjusted OR for ever tobacco chewing was 44.1 (95% CI = 22.0-88.2). An inverse dose-response relationship was seen between BMI and the risk of oral submucous fibrosis when both genders were combined (p for trend = 0.0010), with an OR of 0.5 (95% CI = 0.3-0.9) for the highest BMI quartile compared to the lowest. Alcohol drinking may possibly be associated with the risk of oral submucous fibrosis; the adjusted OR for ever drinking was 2.1 (95% CI = 1.0-4.4). Cigarette smoking did not appear to be a risk factor for women or for men. Both smoking and drinking were rare habits among women. CONCLUSION: This study suggested, for the first time, that BMI was inversely associated with the risk of oral submucous fibrosis for both genders when potential confounding factors were adjusted. Our results indicated that alcohol drinking might be a moderate risk factor and confirmed the previous observation that chewing tobacco was a strong risk factor for oral submucous fibrosis. PMID- 11899120 TI - The greater impact of menopause on ER- than ER+ breast cancer incidence: a possible explanation (United States). AB - OBJECTIVE: Analysis of 3359 Danish breast cancer cases indicated that menopause exerted a greater protective effect on estrogen-receptor negative (ER-) breast cancer than on estrogen-receptor positive (ER+) breast cancer. We examined US age specific breast cancer rates by hormone receptor status in white and black women and men to investigate this unexpected result. METHODS: Age-specific breast cancer incidence rates from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute were analyzed by joint estrogen receptor and progesterone receptor (ER/PR) status of 101,140 white female and 8870 black female cases and by ER status in 706 white male and black male cases diagnosed from 1992 to 1998. Changes in the rate of increase in rates with age were identified using Poisson regression analyses. RESULTS: For both white women and black women the age-specific rates of ER- breast cancer cease increasing after 50 years of age, but age-specific rates of ER+ breast cancer continue to increase after 50 years of age. For men the incidence of ER- cancers may increase at a slower rate than incidence of ER+ cancers in older ages. In women the black rates of ER+ cancers are greater than white rates only until age 35, but black rates of ER- cancers are greater than white rates for all ages. CONCLUSIONS: Differences in age-specific breast cancer incidence patterns by hormone receptor status are similar for black women and white women. The incidence pattern for ER- cancers is consistent with a paracrine model for hormone-stimulated growth in normal breast tissue. The continued increase in ER+ cancers after menopause may be explained by both the paracrine growth model and an increase in the proliferation rate of ER+ cells with age. PMID- 11899121 TI - Sociodemographic predictors of non-attendance at invitational mammography screening--a population-based register study (Sweden). AB - OBJECTIVE: To investigate the role of sociodemographic factors in predicting mammography uptake in an outreach screening program. METHODS: Linkage of data from a regional population-based mammography program with four Swedish nationwide registers: the Population and Housing Census of 1990, the Fertility Register, the Cancer Register, and the Cause of Death Register. We computed odds ratios (OR) and 95% confidence intervals (CI) for non-attendance by sociodemographic factors. Non-attendance was defined as failure to attend in response to the two most recent invitations. RESULTS: Multivariate analyses among 4198 non-attenders and 38,972 attenders revealed that both childless and high-parity women were more likely to be non-attenders (OR = 1.8, 95% CI: 1.6-2.0 and OR = 2.2, 95% CI: 1.8 2.7, respectively). Women living without a partner were less likely to attend (OR = 1.7, 95% CI: 1.5-1.9), as were non-employed women (OR = 2.1, 95% CI: 1.9-2.3). Those renting an apartment were more likely to be non-attenders compared with home-owners (OR = 1.8, 95% CI: 1.6-2.0), and immigrants from non-Nordic countries were more than twice as likely to be non-attenders compared with Swedish-born women (OR = 2.4, 95% CI: 2.0-2.8). CONCLUSIONS: There are identifiable subgroups in which mammography utilization can be increased. Special attention should be paid to women who have never attended, childless women, and non-Nordic immigrants. PMID- 11899122 TI - Hypothesis: does ochratoxin A cause testicular cancer? AB - Little is known about the etiology of testicular cancer, which is the most common cancer among young men. Epidemiologic data point to a carcinogenic exposure in early life or in utero, but the nature of the exposure is unknown. We hypothesize that the mycotoxin, ochratoxin A, is a cause of testicular cancer. Ochratoxin A is a naturally occurring contaminant of cereals, pigmeat, and other foods and is a known genotoxic carcinogen in animals. The major features of the descriptive epidemiology of testicular cancer (a high incidence in northern Europe, increasing incidence over time, and associations with high socioeconomic status, and with poor semen quality) are all associated with exposure to ochratoxin A. Exposure of animals to ochratoxin A via the diet or via in utero transfer induces adducts in testicular DNA. We hypothesize that consumption of foods contaminated with ochratoxin A during pregnancy and/or childhood induces lesions in testicular DNA and that puberty promotes these lesions to testicular cancer. We tested the ochratoxin A hypothesis using ecologic data on the per-capita consumption of cereals, coffee, and pigmeat, the principal dietary sources of ochratoxin A. Incidence rates for testicular cancer in 20 countries were significantly correlated with the per-capita consumption of coffee and pigmeat (r = 0.49 and 0.54, p = 0.03 and 0.01). The ochratoxin A hypothesis offers a coherent explanation for much of the descriptive epidemiology of testicular cancer and suggests new avenues for analytic research. PMID- 11899123 TI - Hypertrophic scar formation following carbon dioxide laser ablation of plantar warts in cyclosporin-treated patients. AB - We present four renal transplant patients who developed hypertrophic scars following carbon dioxide laser ablation of recalcitrant plantar warts. All of the patients were on long-term treatment with cyclosporin, which we believe to be responsible. We discuss several possible mechanisms by which cyclosporin may influence wound healing and scarring. PMID- 11899124 TI - Vitiligo-like leucoderma during photochemotherapy for mycosis fungoides. AB - We describe four patients with mycosis fungoides (MF) in whom depigmentation, and also leucotrichia in one, occurred following the resolution of the eruption during phototherapy (psoralen plus ultraviolet A treatment in three patients, climatotherapy in one). In all cases, the depigmentation was localized to the area of the pre-existing MF lesions. There was no clinically obvious phototoxicity. Biopsy study including S100 staining in all cases, and electron microscopy in one case, demonstrated the total absence of melanocytes, with no evidence of MF. It is suggested that the phototherapy may have activated a cell mediated immunity leading to destruction of the melanocytes. We recommend that vitiligo-like leucoderma be added to the list of untoward effects of phototherapy in MF. PMID- 11899125 TI - Generalized vitiligo after lymphocyte infusion for relapsed leukaemia. AB - Vitiligo is an autoimmune disease caused by T-lymphocyte-mediated destruction of melanocytes. We describe two patients with generalized vitiligo caused iatrogenically after donor lymphocyte infusion (DLI) for leukaemia relapse over 3 years after bone marrow transplantation (BMT). Neither the sibling donor nor the recipient had vitiligo or other autoimmune diseases, and vitiligo did not occur after the first BMT. DLI was accompanied by skin graft-versus-host disease in both cases, which was controlled with immunosuppression. However, over several months, progressive generalized and persistent skin depigmentation occurred in both patients. Peripheral blood molecular studies showed the complete disappearance of host haematolymphopoiesis. The specific destruction of melanocytes in both patients was therefore probably mediated by new alloreactive lymphocytes infused from the donors. PMID- 11899126 TI - Eumycetoma due to Madurella mycetomatis acquired in Jamaica. AB - We report a case of eumycetoma due to Madurella mycetomatis affecting the left dorsal forefoot in a 35-year-old woman. She had spent her childhood in Jamaica, and had been resident in the U.K. for 20 years prior to her presentation. She gave a highly suggestive history for a mycetoma, having observed the intermittent discharge of black grains from the lesion. The diagnosis was confirmed by histological and mycological analysis of the grains, and a magnetic resonance imaging scan excluded osteomyelitis. She has responded very favourably to treatment with itraconazole. Mycetomas due to M. mycetomatis have not previously been reported from Jamaica. PMID- 11899127 TI - Successful topical immunotherapy of bowenoid papulosis with imiquimod. PMID- 11899128 TI - The eclipse naevus: tan centre with stellate brown rim. PMID- 11899129 TI - Acute generalized exanthematous pustulosis induced by icodextrin. PMID- 11899130 TI - The effect of treatment on serum levels of soluble intercellular adhesion molecules and tumour necrosis factor-receptor 1 in psoriasis. PMID- 11899132 TI - Why ultraviolet protection with clothing makes sense. PMID- 11899131 TI - Assessment of acitretin-treated female patients of childbearing age and subsequent risk of teratogenicity. PMID- 11899133 TI - Melanoma tumour markers S100B and MIA: evaluation of stability in serum and blood upon storage and processing. PMID- 11899134 TI - Decreased expression of fas (APO-1/CD95) on lesional CD4+ T lymphocytes in cutaneous T cell lymphomas: correlations with blood data. PMID- 11899136 TI - Pubic trichotillomania in an adult man. PMID- 11899135 TI - Multicentric reticulohistiocytosis associated with idiopathic myelofibrosis. PMID- 11899137 TI - Antibodies targeting extractable nuclear antigens: historical development and current knowledge. PMID- 11899138 TI - Alterations in molecular killing mechanisms: implications in skin disease. AB - Cytolytic T lymphocytes exert two main specific molecular killing mechanisms against target cells, namely (i) they can synthesize and release soluble cytolytic factors, and (ii) they can express effector molecules that act as ligands of receptors expressed by target cells on the cell surface; by these two pathways cytolytic T lymphocytes kill several targets, e.g. cells infected with intracellular pathogens, cells transformed by malignancy and cells producing autoantibodies. This review investigates the contribution from alterations in these molecular killing mechanisms to the pathogenesis of cutaneous diseases. In fact, molecular components involved in such killing mechanisms are often altered or distorted in skin pathology, e.g. cutaneous viral infections, skin cancer, contact hypersensitivity and autoimmune diseases with cutaneous involvement. Treatments capable of repairing the molecular components operating in such killing mechanisms could presumably favour the resolution of these skin diseases. PMID- 11899139 TI - Guidelines for care of contact dermatitis. AB - These guidelines for the management of contact dermatitis have been prepared for dermatologists on behalf of the British Association of Dermatologists. They present evidence-based guidance for treatment, with identification of the strength of evidence available at the time of preparation of the guidelines, including details of relevant epidemiological aspects, diagnosis and investigation. PMID- 11899140 TI - A systematic review of antistreptococcal interventions for guttate and chronic plaque psoriasis. AB - BACKGROUND: Guttate psoriasis is closely associated with preceding or concurrent streptococcal infection. Some authorities have claimed that chronic plaque psoriasis may also be made worse by infection. In view of this many dermatologists have recommended using antibiotics for psoriasis, particularly guttate type. Some dermatologists have also recommended tonsillectomy for psoriasis in patients with recurrent streptococcal pharyngitis. OBJECTIVES: This review aims to assess the evidence for the effectiveness of antistreptococcal interventions, including antibiotics and tonsillectomy in the management of acute guttate and chronic plaque psoriasis. METHODS: Studies were identified by searching the Cochrane Clinical Trials Register (Cochrane Library, Issue 3, 1999), Medline (1966-September 1999), Embase (1988-September 1999), the Salford Database of Psoriasis Trials (to November 1999) and the European Dermato Epidemiology Network (EDEN) Psoriasis Trials Database (to November 1999) for terms (STREPTOCOCC* or ANTIBIOTIC* or TONSIL*) and PSORIASIS using the Cochrane Skin Group search strategy. RESULTS: Only one trial met the selection criteria. This compared the use of two oral antibiotic schedules in 20 psoriasis patients, predominantly of guttate type, who had evidence of beta-haemolytic streptococcal colonization. Either rifampicin or placebo was added to the end of a standard course of phenoxymethylpenicillin or erythromycin. No patient in either arm of the study improved during the observation period. No randomized trials of tonsillectomy for psoriasis were identified. CONCLUSIONS: Although both antibiotics and tonsillectomy have frequently been advocated both for patients with guttate psoriasis and for selected patients with chronic plaque psoriasis, there is to date no good evidence that either intervention is beneficial. PMID- 11899141 TI - A systematic review of treatments for guttate psoriasis. AB - BACKGROUND: Many different therapies are available for treating guttate psoriasis; however, there appears to be little objective evidence for their efficacy OBJECTIVES: This review aims to assess the evidence for the effectiveness of treatments for guttate psoriasis. Antistreptococcal interventions for guttate psoriasis are addressed in a separate review. METHODS: Studies were identified by searching the Cochrane Clinical Trials Register (Cochrane Library, Issue 3, 1999), Medline (1966-September 1999), Embase (1988 September 1999), Salford Database of Psoriasis Trials (to November 1999) and the European Dermato-Epidemiology Network (EDEN) Psoriasis Trials Database (to November 1999) for terms GUTTATE and PSORIASIS. We also searched 100 unselected randomized controlled trials of psoriasis therapy and all 112 randomized controlled trials of phototherapy for psoriasis in the Salford Database of Psoriasis Trials for separate stratification of guttate psoriasis. RESULTS: No published report could be found to support or to challenge current commonly used methods of management. Only one trial that met the selection criteria was identified. In this small study of 21 hospitalized patients with guttate psoriasis, intravenous infusion of an n-3 fatty acid rich lipid emulsion was compared with placebo emulsion containing n-6 fatty acids. The n-3 preparation appeared to be of some benefit for patients with guttate psoriasis. CONCLUSION: There is currently no firm evidence on which to base treatment of acute guttate psoriasis. Studies comparing standard treatment modalities, including phototherapy and topical regimens, are required to enable informed decisions on treatment choices to be made. PMID- 11899142 TI - The mechanism of hyperpigmentation in seborrhoeic keratosis involves the high expression of endothelin-converting enzyme-1alpha and TNF-alpha, which stimulate secretion of endothelin 1. AB - BACKGROUND: Seborrhoeic keratosis (SK) is a benign epidermal tumour with increased pigmentation. We have recently demonstrated that increased secretion of endothelin (ET)-1, a strong keratinocyte-derived mitogen and melanogen for human melanocytes, is intrinsically involved in the hyperpigmentation mechanism of SK. OBJECTIVES: To examine whether the increased ET secretion results from cytokines that induce ET production and/or from differences in the processing of ET that lead to its final active, secreted form. METHODS: We used immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) to determine whether ET-inducing enzymes and/or cytokines are also highly expressed in SK. RESULTS: RT PCR of mRNAs encoding interleukin (IL)-1alpha, tumour necrosis factor (TNF)-alpha and endothelin-converting enzyme (ECE)-1alpha demonstrated that there is an increased expression of TNF-alpha and ECE-1alpha mRNAs in SK, whereas the IL 1alpha transcript is rather downregulated in SK compared with that in perilesional normal epidermis. In parallel, immunohistochemical analysis of SK revealed marked immunostaining for TNF-alpha in basaloid cells at lower levels of the epidermis and in basal cells, and for ECE-1alpha in most basaloid and basal cells in comparison with their weak staining throughout the epidermis in perilesional normal controls. In contrast, immunostaining for IL-1alpha was almost negative in SK relative to distinctive staining throughout the epidermis in the perilesional normal controls. CONCLUSIONS: These findings suggest that the increased secretion of ET-1 leading to enhanced pigmentation in SK results from the co-ordinated increased expression of TNF-alpha and ECE-1alpha. PMID- 11899143 TI - Can we use video images to triage pigmented lesions? AB - BACKGROUND: Advances in telemedicine permit consultations where the doctor and patient are at different sites. OBJECTIVES: To determine whether this technology could be used to triage referrals of pigmented lesions to a dermatology out patient clinic, and thereby assist in managing the ever increasing number of lesions being referred. METHODS: When patients attended clinic, a clinical diagnosis of their pigmented lesion was made and recorded. A still image of each lesion was subsequently taken from a video camera using a PC card, and stored. These images were subsequently viewed in conjunction with the general practitioner's referral information and designated as 'warrants referral' or 'does not warrant referral'. For each lesion this decision was compared with the clinical diagnosis made during the live consultation in the clinic (the 'gold standard'). Clinical diagnoses designated as warranting referral were malignant melanoma, basal cell carcinoma, keratoacanthoma, atypical naevus and pyogenic granuloma (due to the potential clinical confusion with amelanotic melanoma). Lesions that were not considered to warrant referral included benign melanocytic naevus, seborrhoeic keratosis, dermatofibroma, congenital naevus, solar lentigo, actinic keratosis and various other benign conditions. RESULTS: In total, 819 lesions were evaluated, resulting in a mean sensitivity of 81% and specificity of 73% for the technique. CONCLUSIONS: We feel that the overall sensitivity of 81% is encouraging as regards the use of such a technique as a triage tool, but that the inability to examine the whole patient or palpate the lesions is a major drawback in the safe triage of patients with pigmented lesions. PMID- 11899144 TI - Patient satisfaction with teledermatology is related to perceived quality of life. AB - BACKGROUND: There is a lack of good data about patient satisfaction with teledermatology and about its potential interaction with quality-of-life factors. OBJECTIVES: To assess the association between perceived skin-related quality of life and patient satisfaction with a nurse-led teledermatology service. METHODS: In a mobile nurse-led teledermatology clinic located in four inner city general practices in Manchester, the teledermatology service used digital cameras to capture and store images of skin conditions for remote diagnosis by dermatologists. One hundred and twenty-three adult patients, non-urgent dermatology referrals from primary care, completed the Dermatology Life Quality Index (DLQI) and a 15-item patient satisfaction questionnaire. RESULTS: In common with other studies of patient satisfaction, subjects reported highly favourable views of 'hotel' aspects of the service (93%) and found it 'convenient' (86%). However, 40% of patients would have preferred to have had a conventional face-to face consultation with a dermatologist, and 17% felt unable to speak freely about their condition. Patient satisfaction with the service was related to quality of life. Patients reporting lower quality of life as measured by the DLQI were more likely to prefer a face-to-face encounter with a dermatologist (r = 0.216, P < 0.05), and to evince anxiety about being photographed (r = 0.223, P < 0.05). CONCLUSIONS: Patient acceptance and satisfaction with telemedicine services is complicated by patients' subjective health status. Telehealthcare providers need to recognize that patients with poor quality of life may want and benefit from face-to-face interaction with expert clinicians. PMID- 11899145 TI - Numerical, morphological and phenotypic changes in Langerhans cells in the course of murine graft-versus-host disease. AB - BACKGROUND: In the course of graft-versus-host disease (GVHD) or diseases that histologically mimic GVHD (e.g. toxic epidermal necrolysis, Stevens-Johnson syndrome), it is known that epidermal Langerhans cells (LCs) are depleted from the epidermis. However, the mechanism and significance of LC depletion is not well known. OBJECTIVES: To investigate the numerical, morphological and phenotypic changes in LCs and apoptosis of LCs in the course of GVHD using a non irradiated mouse GVHD model. METHODS: BALB/c nu/nu mice and C57BL/6 mice were used as recipients and donors, respectively. Recipient mice were injected with T cell-enriched donor spleen cells. Skin samples were harvested at various times after the inoculation. The numerical and morphological changes were examined by an immunofluorescence study of epidermal sheets. Apoptosis was studied by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling method and flow cytometric analysis using annexin V. Phenotypic change was studied by flow cytometric analysis of epidermal cell suspensions. The mixed epidermal cell lymphocyte reaction (MELR) was performed to examine functional changes in the epidermal cells. RESULTS: Five days after inoculation, a graft versus-host reaction occurred. Epidermal LCs began to decrease from the sixth day. On the fifth day, the LCs became larger and had prominent dendrites. Immediately before the LCs began to decrease, many LCs became round in shape, with scanty dendrites. LC apoptosis was not observed in the epidermis either on the fifth or seventh day. Phenotypically, the expression of CD40, CD80, CD86 and major histocompatibility complex class II antigen on the LCs was upregulated on the fifth and seventh day. Epidermal cells from GVHD mice showed an increased allostimulatory capacity in the secondary MELR. CONCLUSIONS: These results suggest that at early GVHD onset, most LCs may not undergo apoptosis in the epidermis but are phenotypically activated, resulting in further activation of alloreactive T cells and aggravation of the disease. PMID- 11899146 TI - Proapoptotic and antiapoptotic markers in cutaneous T-cell lymphoma skin infiltrates and lymphomatoid papulosis. AB - BACKGROUND: In cutaneous T-cell lymphoma (CTCL) lesions, both reactive T cells and malignant T cells intermingle. The disease progression is mostly slow. Recent evidence suggests that even if clinical remission is reached, malignant cells persist and a relapse follows sooner or later. To wha extent tumour cell apoptosis occurs in the skin lesions either due to the reactive T cells or t therapeutic efforts is not known. OBJECTIVES: To determine the extent of tumour cell apoptosis and the expression of proapoptotic an antiapoptotic markers in serial skin lesion samples from patients with CTCL, and to compare th findings with those in patients with lymphomatoid papulosis (LyP). METHODS: Thirty-four skin samples were obtained from 12 patients with CTCL at the time o diagnosis and at a mean of 1.6, 3 and 6 years later. The patients received psoralen plus ultraviolet (PUVA), electron beam or cytostatic treatments. In addition, fresh post-treatment samples fro three patients with CTCL undergoing PUVA therapy were obtained. For comparison, skin biopsies o five patients with LyP were studied. Immunohistochemical demonstration of the expression of th following markers was performed on formalin-fixed skin sections: Fas (CD95), Fas ligand (FasL) bcl-2, granzyme B, the tumour-suppressor protein PTEN and the effector caspase, caspase 3. Th malignant cells were identified morphologically, and apoptotic cells were identified with th terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling method on parallel sections. RESULTS: In untreated CTCL lesions, apoptotic lymphocytes were extremely rare, and no increase in the number of apoptotic cells was observed after any of the treatments used. In LyP, apoptotic cell were more frequent, comprising on average 5% of the infiltrate. The apoptosis-associated marker Fas, FasL, caspase-3 and granzyme B were expressed by morphologically neoplastic cells in CTCL and by large atypical cells in LyP, with no significant differences. However, only a few reactive cell in CTCL infiltrates expressed granzyme B while about 10% of the corresponding cells were positive in LyP. The expression of antiapoptotic bcl-2 was more frequent in CTCL than in LyP, while PTE expression was high in both instances. The number of bcl-2 + cells tended to decrease after therapy When comparing the findings between the first and the last samples, a decrease in the number of bcl-2+ cells and an increase in Fas+ cells was associated with disease progression, despite therapy, while the opposite was true for remissions. CONCLUSIONS: Apoptosis was found to be a rare event in CTCL lesions irrespective of precedin therapy During patient follow-up, no significant differences in the expression of apoptotic marker was observed while in most cases a lower level of antiapoptotic bcl-2 expression was observed after all types of therapies and in association with disease progression when compared with high expression in the untreated lesions. The absence of apoptosis and high expression of bcl-2 together with a low expression of apoptosis-inducing granzyme B in the reactive lymphocytes in CTC could explain the chronic nature of the disease and the poor response to therapy, while th more frequent occurrence of granzyme B and apoptosis together with a lower level of expressio of bcl-2 by the large atypical cells in LyP could contribute to the favourable outcome of the latter. PMID- 11899147 TI - Cutaneous elimination of 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - BACKGROUND: After exposure, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is excreted via the faeces, breast milk and epidermal lipids. OBJECTIVES: To determine to what extent TCDD is eliminated via the skin and to evaluate whethe cutaneous elimination can be accelerated by the application of petrolatum. METHODS: In two patients severely intoxicated with TCDD, material obtained from the skin surface and, in one patient, cerumen and the content of epithelial cysts, was analysed for TCDD. RESULTS: The TCDD concentration in the initial blood sample taken was 144 000 pg g(-1) blood fa in patient 1, and 26 000 pg g( 1) blood fat in patient 2. Six months later, when the skin tests were performed, the blood TCDD levels had decreased to 80 900 and 16 100 pg g(-1) blood fat, respectively. In the two samples of pooled cyst contents from patient 1, TCDD levels of 34 400 an 18 600 pg g(-1) fat were found. A cerumen sample contained TCDD at 20 500 pg g(-1) fat. In the material collected from the skin surface we observed a linear increase of the amount of TCD measured per test field with time, indicating a continuous elimination of TCDD via the skin. Th daily amount of TCDD eliminated via the skin was 1.51 pg cm(-2) in patient 1 and 0.57 pg cm( 2) in patient 2. Application of petrolatum led to a twofold increase in the amount of TCDD measured in patient 1, but had no significant effect in patient 2. CONCLUSIONS: In our patients, elimination of TCDD via the skin, most probably through desquamating scales, represented 1-2% of the overall daily TCDD elimination rate, with regard to the body surface and when calculated on the basis of the half-life of TCDD at the time of the skin test. If a more typical overall elimination half-life of 7 years is used as the basis for the calculatio the skin would account for 9% (patient 1) and 15% (patient 2) of the overall elimination. Although we observed an increase in TCDD in material derived from the skin surface of up to 100% after application of petrolatum in patient 1, such an approach appears not to be a feasible means to increase elimination. Owing to the small amount of TCDD measured in skin-surface material, as well as in the cyst contents and cerumen obtained from one patient, contamination of the environment and other persons appears highly unlikely. PMID- 11899148 TI - Pituitary-adrenal function following dexamethasone-cyclophosphamide pulse therapy for pemphigus. AB - BACKGROUND: Systemic corticosteroid therapy is known to lead to pituitary-adrenal (PA) suppression. Although patients treated for pemphigus with dexamethasone cyclophosphamide pulse (DCP) therapy have shown no evidence of PA suppression, no study has been conducted to study this possible side-effect of DCP therapy. OBJECTIVES: To study the effect of DCP therapy for pemphigus on PA function. METHODS: A cross-sectional study was carried out in 33 patients who completed phase II of DCP therapy in a single centre, a large teaching hospital in New Delhi, India. Serum cortisol levels following adrenocorticotrophic hormone stimulation were analysed in 31 patients. RESULTS: Seventeen (55%) patients showed suppressed PA function. Suppressed function occurred more frequently in patients who had received pulses of dexamethasone in addition to DCP therapy in the first phase of treatment (P = 0.055). CONCLUSIONS: Suppressed PA function occurs in about half of patients who receive DCP therapy for pemphigus. These patients probably do not require routine replacement therapy with corticosteroids but may need supplementation during periods of stress. PMID- 11899149 TI - Basal cell carcinoma follow-up practices by dermatologists: a national survey. AB - BACKGROUND: After treatment of a basal cell carcinoma (BCC) patients are at risk of recurrence of that BCC; also, patients who have had a primary BCC are those who have an increased risk of developing a subsequent primary BCC. However, long term hospital-based follow-up of all patients would put large strains on the U.K. health service. OBJECTIVES: To investigate the follow-up intentions of U.K. dermatologists for well-defined facial BCC and to investigate the effect that variations in site and clinical indicators might have on those intentions. METHODS: A self-completion questionnaire relating to BCC follow-up sent to 388 dermatology consultants and associate specialists in the U.K. had a response rate of 68%. The effects of treatment modality, tumour site, histology, multiple lesions and various patient variables that might alter the likelihood of follow up were examined. General views on the subject of BCC follow-up were sought. RESULTS: Twenty-seven per cent of respondents reported that they would not review further after excision of a 'well-defined' BCC from inside a central 'T' area on the face; 37% reported that they would review on one occasion; and 36% reported that they review more than once. CONCLUSIONS: While it is currently not feasible to follow-up all treated BCCs, a strategy to identify and monitor high-risk patients and a system to gather long-term outcome data prospectively are necessary aspects of a national health service. This study illustrates that the first issue is being addressed to some extent, but at the currently reported level of BCC follow-up in the U.K. there is little scope for collecting comprehensive long-term data on outcomes. PMID- 11899150 TI - Heterogeneity within tissue-specific macrophage and dendritic cell populations during cutaneous inflammation in atopic dermatitis. AB - BACKGROUND: Macrophages and dendritic cells may play a role in chronicity of atopic dermatitis (AD); however, so far only limited data are documented on the distribution of these cells in the skin during cutaneous inflammation. OBJECTIVES: To gain better insight into the presence and distribution of macrophage and dendritic cell (sub)populations in acutely and chronically inflamed skin of AD patients. METHODS: Chronic inflammatory reactions were studied in lesional AD skin biopsies; the atopy patch test was used as a model for the initiation of AD lesions, representing acute inflammation. To determine the number and phenotype of different dermal macrophage and dendritic cell populations immunohistochemistry and digital imaging were used. RESULTS: There was an increase in macrophage numbers in acutely and chronically inflamed AD skin, whereas absolute dendritic cell numbers were unchanged, compared with non lesional AD skin. Furthermore, phenotypically heterogeneous and overlapping macrophage and dendritic cell populations were present in inflamed AD skin. The classic macrophage marker CD68 and prototypic dendritic cell marker CD1a could bind to the same cell subpopulation in the dermis of inflamed AD skin. Mannose receptors were expressed mainly by macrophages in inflamed AD skin. CONCLUSIONS: In this study we observed changes in macrophage number and phenotype during cutaneous inflammation in AD. Dendritic cell numbers did not change; however, phenotypically dendritic cell and macrophage subpopulations showed increasing overlap during inflammation in AD skin. We show for the first time that within tissue-specific macrophage populations further subpopulations are present, and that monocyte-derived cells may express markers for both dendritic cells and macrophages. Our results point to the existence of a heterogeneous pool of macrophage/dendritic cell-like cells, from which subpopulations of dermal macrophages and dendritic cells arise. PMID- 11899151 TI - Trends in the prevalence of atopic dermatitis in school children: longitudinal study in Osaka Prefecture, Japan, from 1985 to 1997. AB - BACKGROUND: The authors have carried out questionnaire surveys of health symptoms in school children (aged 7-12 years) in Osaka Prefecture, Japan. In this study, the geographical distribution of and trends in the prevalence of atopic dermatitis were evaluated. OBJECTIVES: To assess the relationship between the prevalence of atopic dermatitis in children and the environmental factors, and to estimate future changes of the prevalence of atopic dermatitis. METHODS: A total of seven population surveys were carried out at 2-year intervals between 1985 and 1997 in a total of about 4 million primary school children (460 000-740 000 per survey). RESULTS: The lifetime prevalence of atopic dermatitis increased from 15.0% in 1985 to 24.1% in 1993 but levelled off thereafter. According to the school year, the prevalence was higher in lower-year pupils. However, the prevalence stopped rising earlier in lower-year pupils and began to decrease in 1997 in first- to third-year pupils. The prevalence according to areas was significantly correlated negatively with air pollution and positively with the income index. According to the income classes, the prevalence increased until 1993 but levelled off thereafter in all classes, which reduced the differences among the classes. CONCLUSIONS: The increase of the prevalence of atopic dermatitis in school children stopped in 1993 in Osaka Prefecture, Japan. It is conceivable that the prevalence had reached the saturation level. PMID- 11899153 TI - Risk of second primary malignancies in patients with cutaneous melanoma. AB - BACKGROUND: In several studies an increased risk for development of breast cancer, malignant lymphoma and neoplasms of the kidney as second primary cancers in patients with cutaneous melanomas was discussed. OBJECTIVES: To determine the risk for development of second primary neoplasms in patients with cutaneous melanomas. METHODS: A prospective study was performed between 1977 and 1992 to evaluate the occurrence of second primary malignancies in 4597 patients (2083 men, 2514 women) with invasive cutaneous melanomas, diagnosed and treated at the Department of Dermatology and Allergology, Ludwig-Maximilians-University, Munich, Germany. RESULTS: During a median follow-up of 7.2 years, 296 of 4597 patients (6.4%) developed one or more neoplasms at the time of or subsequent to the diagnosis of the first cutaneous melanoma. More than half of these patients developed one or more further melanomas (152, 3.3%). Cancers of the breast, prostate, colon, rectum and kidney occurred less frequently. Statistical calculations revealed a 33.8-fold increased risk for the development of a second melanoma in the entire group [relative risk 38.5 for men (95% CI, 30.4-48.1), 29.0 for women (95% CI, 22.0-37.5)]. Moreover, a significantly increased risk for the development of kidney carcinoma in men was found [relative risk 3.5 (95%, CI, 1.4-7.2)]. CONCLUSIONS: Thorough follow-up and skin examination in patients with cutaneous melanomas is recommended for early detection of other primary melanomas. Furthermore, ultrasound examinations routinely performed in melanoma patients for the detection of melanoma metastases may also be of value for early detection of kidney carcinomas in male patients. PMID- 11899152 TI - Mycosis fungoides: HLA class II associations among Ashkenazi and non-Ashkenazi Jewish patients. AB - BACKGROUND: An immunogenetic mechanism has been suggested to play a role in the pathogenesis of mycosis fungoides (MF). While results of studies on HLA class I associations haveproved inconsistent, two previous studies showed that certain HLA class II alleles were significantly increased among North American caucasian patients with MF: HLA-DRB1*11 and DQB1*03. OBJECTIVES: To investigate the possible HLA class I and class II associations with MF among Jewish patients. METHODS: The patient group comprised 68 Jewish patients with MF: 38 Ashkenazi and 30 non-Ashkenazi. The control group comprised 252 healthy Jewish volunteers: 132 Ashkenazi and 120 non-Ashkenazi. Tissue typing for HLA class I (A and B) was performed using the National Institutes of Health microlymphocytotoxicity technique. DNA-based low-medium resolution analysis for DRB1* and DQB1* alleles was performed using polymerase chain reaction (PCR) amplification with sequence specific primers. For those alleles found to have significantly increased frequency, high-resolution analysis was done by means of PCR sequence-specific oligotyping. RESULTS: The allele frequency of HLA-DRB1*11 was found to be significantly increased but only among Ashkenazi patients with MF (30% vs. 19% in the controls; P = 0.034). High-resolution analysis for DRB1*11, not previously performed, suggested that its greater frequency is due to the increased number of Ashkenazi MF patients with the DRB1*1104 allele (P corrected = 0.036). Analysed together, DQB1*03 alleles (DQB1*0301-0304) had a significantly greater frequency in MF as a group as compared with controls (47% vs. 33%, P = 0.003). DQB1*0301 was demonstrated to be the specific allele associated with MF in Jewish patients (allele frequency of 36% vs. 23% in controls; P corrected = 0.0068), which was not the case for North American caucasian patients with MF. No greater frequencies of any of the HLA class I A or B antigens were found. CONCLUSIONS: Our findings further demonstrate the 'universality' of MF HLA class II susceptibility alleles, i.e. HLA-DRB1*11 and HLA-DQB1*03, suggesting that HLA polymorphism is likely to be important in the pathogenesis of MF in Jewish patients, as it is in North American caucasian patients. Not previously reported is our finding that HLA-DRB1*1104 is the specific allele more prevalent among patients with MF. Our study also underscores some differences in HLA profiles between non-Jewish and Jewish patients with MF and between Ashkenazi and non Ashkenazi Jewish patients, indicating the possibility of diverse HLA disease associations in populations with different genetic backgrounds. Our study provides further evidence for the lack of association between HLA class I and MF. PMID- 11899154 TI - False-negative results in immunoblot assay of serum IgA antibodies reactive with the 180-kDa bullous pemphigoid antigen: the importance of primary incubation temperature. AB - BACKGROUND: Different subepidermal autoimmune blistering skin disorders are characterized by linear deposition of IgA, sometimes accompanied by linear IgG, along the epidermal basement membrane zone. Identification of the targeted autoantigen is usually attempted by immunoblotting. Although immunoblotting works well for human IgG, the method is less successful for IgA and often no or only faint signals are obtained. OBJECTIVES: To improve the method of immunoblotting for diagnosis of IgA-mediated bullous dermatoses. METHODS: Eleven sera, selected from patients with linear deposition of IgA along the epidermal basement membrane zone in vivo, were tested by immunoblotting for antigen specificity using different primary incubation temperatures. RESULTS: No reliable information regarding IgA antigen specificity was obtained when the primary incubation was undertaken at room temperature. In 10 of 11 sera, IgA bound to the 180-kDa bullous pemphigoid antigen (BP180) when the primary incubation temperature was increased to 37 degrees C. CONCLUSIONS: Primary incubation at room temperature may result in false-negative results in the IgA-BP180 immunoblot assay. PMID- 11899155 TI - Lack of association between hepatotropic transfusion transmitted virus infection and oral lichen planus in British and Italian populations. AB - BACKGROUND: A possible association between oral lichen planus (OLP) and chronic hepatic disease has been found in some populations, although this is probably geographically influenced. In 1997 a new hepatotropic virus, transfusion transmitted virus (TTV), was identified but has not been studied in relation to OLP. OBJECTIVE: The present investigation evaluated the genoprevalence of TTV DNA in the sera of British and Italian patients suffering from OLP using two different sets of primers to identify TTV subgenomic DNA. METHODS: Study groups comprised 40 adult subjects (21 British, 19 Italian) with OLP. For each country, two control groups, a disease-control group and a healthy-control group, were included. The presence of TTV DNA in the sera of patients and control subjects was assessed using two different polymerase chain reactions (PCR) and DNA sequence analysis. RESULTS: Statistical analysis did not reveal evidence of any association between TTV infection and OLP or country of residence. CONCLUSION: An association between TTV and OLP is unlikely. PMID- 11899156 TI - Bullous pemphigoid in pregnancy: contrasting behaviour in two patients. AB - Bullous pemphigoid (BP) is an increasingly common immunobullous disease of the elderly, and, due to the late age of onset, is rarely seen in women of fertile age. Consequently, to the best of our knowledge no cases of BP in pregnancy have been described. We present two cases of BP that have differed in disease behaviour during pregnancy. PMID- 11899157 TI - Cicatricial pemphigoid with circulating autoantibodies to beta4 integrin, bullous pemphigoid 180 and bullous pemphigoid 230. AB - Cicatricial pemphigoid is a heterogeneous group of autoimmune subepidermal blistering diseases associated most commonly with autoantibodies to bullous pemphigoid (BP)180 and less frequently with those to laminin 5 or type VII collagen. In addition, a few cases have been described with autoantibodies to the beta4 subunit of alpha6beta4 integrin. We describe a patient with extensive disease of ocular, oral, pharyngeal, laryngeal and genital mucous membranes that healed with scarring of conjunctivae. IgG autoantibodies bound to the dermal epidermal junction on direct immunofluorescence (IF) microscopy and to the epidermal side of 1 mol L(-1) NaCl-split skin on indirect IF microscopy. Our patient's circulating IgG recognized a 205-kDa protein in extracts of 293T cells transfected with the beta4 subunit of alpha6beta4 integrin and in the cell extract of DJM-1 cells. Our patient's IgG and IgA autoantibodies also reacted with full-length BP180 derived from epidermal extracts and the ectodomain of BP180 (LAD-1) derived from culture supernatant of keratinocytes. In addition, a weak IgG reaction with BP230 was noted. The disease rapidly responded to dexamethasone-cyclophosphamide pulse therapy, and immunoblot reactivity to both beta4 integrin and BP180 decreased according to disease activity. PMID- 11899158 TI - Avoiding aortic clamping during coronary artery bypass using an automated anastomotic device. AB - In its current application, off-pump coronary artery bypass grafting (OPCAB) requires clamping of the aorta to perform the proximal anastomosis. One of the important theoretical advantages of OPCAB is to avoid the undesirable effects of cross-clamping of the aorta. We report our early experience with a technique of no aortic clamping using the St. Jude aortic connector system in 11 patients. PMID- 11899159 TI - The endothelium in clinical cardiac transplantation. AB - Cardiac transplantation is the most successful therapy for refractory heart failure, but clinical transplantation is still confronted with the problems of acute rejection and acute pump failure. The limiting factor in achieving prolonged survival remains cardiac allograft vasculopathy. In recent years it has become apparent that from brain death onward, the cardiac endothelium plays a key role in these acute and chronic events. Brain death is associated with an inflammatory response that primes the endothelium for cumulative injury during the subsequent stages of ischemic cold storage, reperfusion and allorecognition. As a structural and functional interface, the endothelium is the site at which inflammatory cells move from the bloodstream through the vessel wall into the parenchyma. The endothelium interacts with the complement system, the coagulation and inflammatory cascades, circulating leukocytes, the immune system, the smooth muscle in the vessel wall, and the surrounding matrix and cardiomyocytes. A better understanding of its many roles may lead to expansion of our therapeutic possibilities and better outcomes overall. This article reviews the possible roles of the endothelium in relation to cardiac transplantation, and discusses the diagnostic and therapeutic modalities that are available to date. PMID- 11899160 TI - Congenital Heart Surgery Nomenclature and Database Project: update and proposed data harvest. AB - The first report of The Society of Thoracic Surgeons (STS) National Congenital Heart Surgery Database in 1998 reported the clinical features of 18 congenital heart categories. The report provided a significant amount of important information and also highlighted the strengths and weaknesses of the existing database. Following this report the STS Congenital Heart Surgery Committee in cooperation with the European Association of Cardio-Thoracic Surgery and the European Congenital Heart Surgeons Foundation initiated the International Congenital Heart Surgery Nomenclature and Database Project. The goal was to begin the standardization of nomenclature reporting strategies and establish the foundations for an international congenital heart surgery database. The first report of the International Congenital Heart Surgery Nomenclature Project was published in The Annals of Thoracic Surgery in April 2000. The current report outlines modifications to the minimum dataset as well as the diagnoses and procedure short lists. Plans for the next STS National Congenital Heart Surgery Database harvest are also presented. PMID- 11899161 TI - Implementation of new technology for CABG in low-risk patients: could it be too soon? PMID- 11899162 TI - Triangular resection of prolapsing anterior mitral leaflet. PMID- 11899163 TI - Spinal cord protection during descending aortic aneurysm repair. PMID- 11899164 TI - The wider adoption of minimally invasive valvular heart surgery. PMID- 11899165 TI - Inflammatory response after BioGlue application. PMID- 11899166 TI - Preoperative IABP in high-risk patients reduces postoperative lactate release and subsequent mortality. PMID- 11899167 TI - Problems in the evaluation of thymectomy for myasthenia gravis. PMID- 11899168 TI - Acute type A aortic dissection: can we dramatically reduce the surgical mortality? PMID- 11899169 TI - Gene therapy for thoracic disease: practice, promise, and pragmatism. PMID- 11899171 TI - Factors influencing intensive care unit length of stay after surgery for acute aortic dissection type A. AB - BACKGROUND: Operative mortality after acute aortic dissection type A is still high, and prolonged stay at the intensive care unit is common. Little has been documented about factors influencing the intensive care unit length of stay. The aim of this study was to determine such variables. METHODS: During a 10-year period, 67 patients (47 male, 20 female) were operated on for acute aortic dissection type A. In 42 patients (63%), an ascending aortic replacement was performed, 23 patients (34%) underwent a Bentall procedure, and 2 patients (3%) received a valve-sparing David type of operation. In 14 of these cases (20%), an additional partial or total arch replacement was performed. RESULTS: Hospital mortality was 9 of 67 (14%). Median postoperative intensive care unit length of stay was 5 days (range, 1 to 72 days). Intensive care unit stay was in univariate analysis significantly influenced by the following factors: age (p = 0.008), body mass index (p = 0.039), cardiopulmonary bypass time (p = 0.018), aortic cross clamp time (p = 0.031), postoperative low cardiac output syndrome (p < 0.001), and postoperative lactate levels (p = 0.01). By multivariate analysis, age (p = 0.012), cardiopulmonary bypass time (p = 0.037), and the presence of a postoperative low cardiac output syndrome (p < 0.001) significantly influenced intensive care unit stay. CONCLUSIONS: Stay in the intensive care unit after operation for acute aortic dissection type A seems to be determined by age, cardiopulmonary bypass time, and the postoperative presence of a low cardiac output syndrome. PMID- 11899170 TI - Acute type A dissection: conservative methods provide consistently low mortality. AB - BACKGROUND: A wide spectrum of operative techniques are applied in acute type A dissection. Most convey hospital mortality between 10% and 20%. In this high-risk setting, we believe that a conservative approach to the aortic root and the complete resection of the primary tear are important. We reviewed the results of this policy from our aortic surgery database. METHODS: Between 1988 and December 2000, 95 acute type A dissection patients were operated on by one surgeon. They included 70 men and 25 women aged 37 to 81 years (mean 65 years). Six had Marfan syndrome. Aortic root restoration or replacement was performed during cooling, open arch repair during circulatory arrest, and hemostasis while rewarming. Eighty-seven patients had ascending aortic replacement with glue resuspension of the valve. Two others had had aortic valve replacement previously. Aortic root and partial arch replacement was performed in 6 Marfan patients. Eighteen patients had hemiarch replacement, and 6 had total arch replacement to excise the tear. RESULTS: Five patients died in hospital (5.3% 30-day mortality) and another after early readmission for mediastinal infection (6.3% total mortality). There were no deaths from bleeding. Two patients required aortic valve replacement for aortic regurgitation 2.5 and 3.0 years postoperatively. Two others required total arch replacement and thromboexclusion procedures, respectively. CONCLUSIONS: Our policy of primary tear excision and preservation of the native aortic valve has resulted in low overall mortality. We still prefer to replace the aortic root in dissected Marfan patients. In this high-risk condition, hospital survival is of paramount importance. A conservative "pathology-oriented" approach helps to achieve this aim. PMID- 11899172 TI - Reduction aortoplasty for dilatation of the ascending aorta in patients with bicuspid aortic valve. AB - BACKGROUND: Individuals with bicuspid aortic valve tend to develop a dilatation of the ascending aorta. It is controversial whether the dilated ascending aorta should be replaced with a tube graft or whether the diameter of the aorta should be reduced by reduction aortoplasty. Furthermore, it is unclear whether an external prosthetic support of the reduction aortoplasty is necessary. The aim of this study is to analyze the results of reduction aortoplasty with and without external prosthetic support. METHODS: Between 1985 and 1999, a total of 115 patients with bicuspid aortic valve and dilatation of the ascending aorta underwent reduction aortoplasty in combination with other types of open-heart procedure at our institution. The diameter of the ascending aorta was measured before and early after surgery and then later between 12 and 144 months (mean 40 months) postoperatively using echocardiography and computed tomography. RESULTS: The reduction aortoplasty decreased the internal diameter of the aorta from 48.7+/-5.1 mm preoperatively to 36.9+/-3.6 mm early after surgery (p = 0.0001). During follow-up, there was no increase of the aortic diameter either in patients with external prosthetic support or in 97 of 106 patients without external prosthetic support. The diameter increased only in 9 (8.5%) of 106 patients without external aortic support by 4 to 8 mm. In patients with postoperative diameter increase, the aortic diameter after operation had been higher than in patients without a postoperative increase of the aortic diameter (41.4+/-3.1 mm vs 36.6+/-3.4 mm; p < 0.0001). CONCLUSIONS: Reduction aortoplasty showed good long-term results in patients with bicuspid aortic valve and dilatation of the ascending aorta. Redilation of the aorta occurred only in patients with a suboptimal diameter reduction. PMID- 11899173 TI - Complete aortic root replacement in patients with small aortic annulus. AB - BACKGROUND: We evaluated the effectiveness of our surgical method using a modified self-assembled valved composite graft in patients with a narrow aortic annulus. METHODS: Between August 2000 and May 2001, 10 consecutive patients with a narrow aortic annulus underwent replacement of the aortic valve and the ascending aorta using a valved composite graft with mechanical valve prosthesis. The indication for surgery was aneurysm of the ascending aorta (8 patients) and aortic dissection (2 patients). To avoid valve-patient mismatch, a modified self assembled valved composite graft was used. RESULTS: There was no hospital mortality. Echocardiographic evaluation before discharge showed excellent hemodynamics with a mean transvalvular gradient of 10.7 mm Hg (standard deviation +/- 2.8 mm Hg). CONCLUSIONS: The described valved composite graft offers very good hemodynamic performance and is a simple and effective device to avoid valve patient mismatch in patients with a small aortic annulus who need aortic root replacement. PMID- 11899174 TI - Renal perfusion during thoracoabdominal aortic operations: cold crystalloid is superior to normothermic blood. AB - BACKGROUND: Renal failure remains a common complication of thoracoabdominal aortic aneurysm repair. The purpose of this randomized clinical trial was to compare two methods of selective renal perfusion--cold crystalloid perfusion versus normothermic blood perfusion--and determine which technique provides the best kidney protection during thoracoabdominal aortic aneurysm repair. METHODS: Thirty randomized patients undergoing Crawford extent II thoracoabdominal aortic aneurysm repair with left heart bypass had renal artery perfusion with either 4 degrees C Ringer's lactate solution (14 patients) or normothermic blood from the bypass circuit (16 patients). Acute renal dysfunction was defined as an elevation in serum creatinine level exceeding 50% of baseline within 10 postoperative days. RESULTS: One death occurred in each group. One patient in the blood perfusion group experienced renal failure requiring hemodialysis. Ten patients (63%) in the blood perfusion group and 3 patients (21%) in the cold crystalloid perfusion group experienced acute renal dysfunction (p = 0.03). Multivariable analysis confirmed that the use of cold crystalloid perfusion was independently protective against acute renal dysfunction (p = 0.02; odds ratio, 0.133). CONCLUSIONS: When using left heart bypass during repair of extensive thoracoabdominal aortic aneurysms, selective cold crystalloid perfusion offers superior renal protection when compared with conventional normothermic blood perfusion. PMID- 11899175 TI - Effectiveness of combined blood conservation measures in thoracic aortic operations with deep hypothermic circulatory arrest. AB - BACKGROUND: The effectiveness of blood conservation measures for thoracic aortic operations with deep hypothermic circulatory arrest has not yet been documented. METHODS: From July 1997 to December 2000, 148 thoracic aortic operations were performed in our department. Sixty-one cases involving patients who underwent elective thoracic aortic operation with deep hypothermic circulatory arrest were reviewed retrospectively. RESULTS: Seventeen patients did not meet the criteria for the blood conservation program and were excluded from the present study. Therefore, 44 patients were analyzed in this study. Overall, 50% of patients did not require operative homologous blood transfusion (HBT) and 43% did not require in-hospital HBT. Smaller amounts of autologous donation, greater blood loss, and a longer operation time were independent risk factors for HBT requirement. Among 16 patients who had made an autologous donation of 1,600 mL or greater, 75% did not require intraoperative HBT and 69% did not require in-hospital HBT. The overall perioperative mortality rate was 4.5%. As for postoperative complications, prolonged intubation and postoperative infection were significantly more frequent among patients who required in-hospital HBT. CONCLUSIONS: Our combined blood conservation measures were effective in avoiding HBT during major thoracic aortic operations with deep hypothermic circulatory arrest and may have reduced postoperative complications. The amount of the autologous donation was a strong predictor for avoiding HBT. PMID- 11899176 TI - Right ventricular dysfunction and organ failure in left ventricular assist device recipients: a continuing problem. AB - BACKGROUND: Although right ventricular assist device (RVAD) use has declined with the introduction of inhaled nitric oxide and phosphodiesterase inhibitors (type III), right ventricular dysfunction (RVD) is still a serious problem in patients receiving left ventricular assist devices (LVAD). METHODS: We retrospectively analyzed Thoratec Vented Electrical LVAD recipients between June 1996 and September 1999. RVD was defined as inotropic requirement 14 days or more or need for RVAD postoperatively, or both. RESULTS: Sixty-nine LVAD recipients were analyzed. Twenty-one patients (30.4%) had RVD, with 1 patient requiring RVAD insertion, and there were 48 non-RVD patients. There were no significant differences between both groups for age, sex, etiology of congestive heart failure, days of support, and preoperative hemodynamics. Preoperative right ventricle stroke work index (mm Hg x m(-2) x L(-1)) had a trend toward being lower in the RVD group (4.1+/-3.2 versus 6.1+/-3.7, p = 0.06). A higher preoperative total bilirubin (mg/dL) was noticed in the RVD group (4.0+/-5.2 versus 2.1+/-1.7). The RVD group had a higher postoperative creatinine (2.2+/-1.4 mg/dL versus 1.5+/-0.8 mg/dL), incidence of continuous venovenous hemofiltration dialysis (73% versus 26%), transfusion of packed red blood cells (43.2+/-28.6 units versus 24.7+/-18.9 units), platelets (58.6+/-46.1 units versus 30.2+/-20.4 units), with longer intensive care unit length of stay (33.6+/-34.7 days versus 9.1+/-6.9) and higher mortality (42.8% versus 14.5%). When deaths were excluded, both intensive care unit and postoperative length of stay were significantly longer in the RVD group. CONCLUSIONS: RVD in LVAD recipients remains poorly identified and is associated with a high transfusion rate and end organ failure that results in increased intensive care unit and hospital length of stay, and a high mortality rate. Preoperative identification of risk factors for RVD may select patients who would benefit from a biventricular assist device and prevent the subsequent end organ failure. PMID- 11899177 TI - Reductive annuloplasty of double orifices in patients with primary dilated cardiomyopathy. AB - BACKGROUND: Patients with primary dilated cardiomyopathy exhibit extensive remodeling of the left ventricle, mitral and tricuspid annular dilation and both mitral and tricuspid regurgitation. These factors significantly contribute to heart failure, and are predictors of early lethal outcome. The aim of this study is to show hemodynamic and clinical improvement after reductive annuloplasty of both mitral and tricuspid orifices in primary dilated cardiomyopathy. METHODS: There were 76 patients with primary dilated cardiomyopathy. Mitral annuloplasty using a Carpentier-Edwards sizer was performed on 9 patients, and posterior semicircular reductive annuloplasty was performed on 67 patients. Modified De Vega's tricuspid annuloplasty was performed on all patients. RESULTS: Immediate and long-term results showed significant improvement in hemodynamic values and myocardial contractility after operation. CONCLUSIONS: Reductive annuloplasty of both mitral and tricuspid orifices corrects remodeling of the left ventricle of the heart, changes sphericity and geometry of the left ventricle, improves hemodynamic action of the left and right ventricle, and slows down progression of heart failure. We recommend reductive annuloplasty of both mitral and tricuspid orifices before or soon after the first decompensation. PMID- 11899178 TI - Improvement of tricuspid regurgitation after pulmonary thromboendarterectomy. AB - BACKGROUND: For patients with chronic thromboembolic pulmonary hypertension who undergo pulmonary thromboendarterectomy (PTE) it has not yet been systematically investigated how operation affects the severity of tricuspid regurgitation (TR). This study sought (1) to evaluate the extent of TR reversibility after operation, (2) to identify potential predictors of the reversibility of TR, and (3) to investigate the influence of geometric and hemodynamic alterations on the extent of TR severity. METHODS: Thirty-nine patients (55+/-12 years) undergoing PTE without tricuspid valve repair were investigated before and 13+/-8 days after operation by Doppler color flow mapping. Geometry of the tricuspid valve as well as right ventricular size and function were determined with echocardiography. Mean pulmonary arterial pressure was determined invasively. RESULTS: After PTE, mean pulmonary arterial pressure was significantly lower (48+/-10 versus 25+/-7 mm Hg, p < 0.05). Most of the patients had a distinct reduction of TR, and the improvement trend showed on the severity scale: number of patients with 4+TR (23 -> 4), 3+TR (12 --> 12), 2+TR (2 --> 13), and 1+TR (2 --> 10). Examination after PTE revealed profound reduction of right ventricular size and annulus diameter, with a normalization of the valvular geometry. However, none of the study variables were useful as indicators of the postoperative outcome. CONCLUSIONS: After PTE without additional valve repair most patients show significantly reduced severity of TR soon afterward; the very few cases in which TR does not improve remain unidentifiable before operation. Our recommendation is consequently to refrain from additional tricuspid repair in patients undergoing PTE. PMID- 11899179 TI - Assessment of cardiac performance using Tei indices in patients undergoing pulmonary thromboendarterectomy. AB - BACKGROUND: This study was designed to evaluate left and right ventricular performance using Tei indices in patients with severe chronic thromboembolic pulmonary hypertension undergoing pulmonary thromboendarterectomy (PTE). The Doppler-derived indices are easily measurable indicators of ventricular function based on nongeometric assessment, which helps overcome some of the difficulties entailed in the geometric assessment of left ventricular (LV) and right ventricular (RV) function in pulmonary hypertension. METHODS: The indices were derived for 24 patients (aged 54+/-14 years) before and after PTE. Calculation of these indices was based on the duration of two time intervals using the formula (A - B)/B, where A is the interval between cessation and onset of mitral inflow (or tricuspid inflow) and B is LV or RV ejection time. In addition, LV and RV end diastolic and end-systolic chamber areas were determined using two-dimensional echocardiography, and systolic function was calculated. Mean pulmonary artery pressure was determined invasively. RESULTS: PTE led to a significant reduction of mean pulmonary artery pressure (46+/-10 versus 25+/-6 mm Hg; p < 0.05). LV and RV indices were abnormally high before surgery, declined significantly afterwards, and then almost matched normal values (0.61+/-0.26 versus 0.37+/ 0.18; p < 0.05 and 0.55+/-0.22 versus 0.37+/-0.13; p < 0.05). Geometric assessment of the left and right ventricle also showed impaired systolic function before PTE, with significant improvement after surgery. CONCLUSIONS: LV and RV Tei indices allow a quantitative assessment of ventricular function in patients undergoing PTE. Lower indices after surgery reflect an improvement of the previously impaired cardiac function. Our results emphasize the value of PTE in the treatment of chronic thromboembolic pulmonary hypertension. PMID- 11899180 TI - Are stentless valves hemodynamically superior to stented valves? A prospective randomized trial. AB - BACKGROUND: Although stentless aortic bioprostheses are believed to offer improved outcomes, hemodynamic benefits remain unsubstantiated. METHODS: Fifty three patients were randomized to receive the stented C-E pericardial valve (CE) and 46 patients the Toronto Stentless Porcine valve (SPV). Annuli were sized for the optimal insertion of both valve types, such that surgeons were required to commit to specific valve sizes before randomization. Echocardiographic measurements and functional status (Duke Activity Status Index) were assessed at 3 and 12 months postoperatively. RESULTS: Although cardiopulmonary bypass times (CE: 118.6+/-36.3 minutes; SPV: 148.5+/-30.9 minutes; p = 0.0001) and aortic cross-clamp times (CE: 95.4+/-28.6 minutes; SPV: 123.6+/-24.1 minutes; p = 0.0001) were significantly prolonged in the SPV group, perioperative morbidity and mortality was similar between groups. Neither valve offered a superior internal diameter for any given annular diameter (mean decrease in left ventricular outflow tract diameter after valvular implantation: SPV: 3.4+/-1.11 mm versus CE: 3.7+/-1.33 mm; p = 0.25). Although labeled mean valve size was significantly larger in the SPV group, the actual mean valve size based on internal valvular diameter was no different between groups (CE: 21.9+/-2.0 mm; SPV: 22.3+/-2.0 mm; p = 0.286). Although effective orifice areas increased, and mean and peak transvalvular gradients decreased in both groups over time, no differences were demonstrated between groups at 12 months. Similarly, although significant regression of left ventricular mass was accomplished in both groups over time, no differences were demonstrated between groups. Finally, Duke Activity Status Index scores of functional status improved in both groups over time; however, no differences were noted between groups at 12 months postoperatively. CONCLUSIONS: Although offering excellent outcomes, stentless valves did not demonstrate superior hemodynamic indices in comparison to stented valves up to 12 months after implantation. PMID- 11899181 TI - Aortic valve surgery after previous coronary artery bypass grafting with functioning internal mammary artery grafts. AB - BACKGROUND: Aortic valve surgery after coronary artery bypass grafting (CABG) in the setting of patent pedicled internal mammary artery (IMA) grafts poses a high risk because of the underlying ischemic and valve disease. Unlike mitral valve surgery or CABG, in which aortic clamping (AoX) may be optional, aortic valve surgery uniformly requires AoX unless circulatory arrest is used. Management of the IMA graft in these circumstances has traditionally involved dissection and clamping to prevent regional myocardial warming and cardioplegia "washout" during AoX. An alternative strategy involves avoiding dissection of the IMA, leaving the IMA graft open and establishing moderate-to-deep hypothermia during AoX and cardioplegic arrest. To date, no study has been published documenting the safety and efficacy of the latter practice. METHODS: A total of 94 patients who had patent IMA graft and underwent aortic valve surgery under AoX and cardioplegia between April 1992 and March 2001 were analyzed. The IMA was avoided and left open during AoX, and the patients were cooled systemically (median 20 degrees C). Patients ranged in age from 55 to 90 years (median 73.5 years). Ejection fraction was 15% to 83% (median 50%). Of the patients, 18 (19%) underwent minimally invasive upper hemi-resternotomy. Analysis for predictors of outcome was performed. RESULTS: The operative mortality, perioperative myocardial infarction (MI), and stroke rates were 6.4%, 7%, and 11%, respectively. No significant independent predictors of operative mortality or MI could be identified in the multivariate analysis, although a trend was shown for operative mortality with urgent procedures and patients requiring concomitant surgery of the ascending or arch aorta or aortic root. Advanced age and prolonged cardiopulmonary bypass predicted stroke in the multivariate analysis. There were five (5%) IMA injuries, all occurring during reentry or mediastinal dissection, but none in the subgroup of patients who underwent minimally invasive procedures. All patients survived. CONCLUSIONS: Patients undergoing aortic valve surgery after CABG in the presence of patent IMA represent a potentially high-risk group. Because AoX is almost uniformly required, a decision regarding the management of the IMA pedicle is needed. We have found that leaving the IMA undissected and unclamped is a reasonable strategy, provided that systemic cooling for myocardial protection is established to prevent regional warming and to compensate for cardioplegia washout effect during AoX. PMID- 11899182 TI - Safety and efficacy of one stage off-pump coronary artery operation and carotid endarterectomy. AB - BACKGROUND: Patients with concomitant occlusive disease of coronary and carotid arteries remain at high risk of perioperative stroke and myocardial infarction. Combined coronary artery bypass grafting on cardiopulmonary bypass and carotid endarterectomy has been shown to give good results for this category of patients. In the present study, we analyzed our results of off-pump coronary artery bypass grafting and carotid endarterectomy as a one-stage procedure. METHODS: Between January 1997 and December 2000, 82 patients underwent combined off-pump coronary artery bypass grafting and carotid endarterectomy. All patients were evaluated by preoperative carotid duplex scanning and carotid angiography. All patients had more than or equal to 70% carotid artery stenosis. There were 35 asymptomatic patients (42.7%) and 47 symptomatic patients (57.3%). Carotid endarterectomy was performed before coronary artery bypass grafting in all the patients. RESULTS: There were 66 males (80.5%) and 16 females (19.5%) with a mean age of 63+/-8 years. The average number of grafts was 3.4+/-0.8. There was no hospital mortality. One patient had perioperative myocardial infarction. None of the patients had stroke. One patient had transient neurologic deficit and 1 patient had temporary 12th nerve dysfunction; both recovered completely. There was no incidence of neck wound infection, although 1 patient developed neck hematoma that required reexploration. At a mean follow-up of 2.2+/-0.7 years, 1 patient required contralateral carotid endarterectomy and 1 patient died because of cardiac failure. CONCLUSIONS: Combined off-pump coronary artery bypass grafting and carotid endarterectomy is a safe and effective procedure in patients with significant concomitant carotid and coronary artery disease. PMID- 11899183 TI - Conversion to off-pump coronary bypass without increased morbidity or change in practice. AB - BACKGROUND: This article examines the feasibility of complete conversion from conventional coronary artery operation to routine off-pump coronary bypass operation. METHODS: Data on our first 285 off-pump procedures using the Octopus system (Medtronic Inc, Minneapolis, MN) represent our learning curve. This is a complete experience in coronary bypass surgery over 16 months. RESULTS: The cohort was nonselected. All patients had at least two-vessel disease. Eight hundred seven grafts were performed (mean, 2.8 per patient) of which 647 grafts (84%) were arterial (mean, 2.3 per patient). One hundred seventy nine patients (63%) underwent total arterial revascularization. Eight patients required cardiopulmonary bypass; all other operations were completed off-pump. Complications were: mortality, 3 patients (1.5%); renal failure, 24 patients (8%); stroke, 2 patients (< 1%); and atrial fibrillation, 60 patients (21%). The morbidity data and frequency of arterial grafting did not differ from that of 355 patients who underwent coronary bypass operations in a preceding 18-month period. CONCLUSIONS: Complete shift from routine use of cardiopulmonary bypass to nonselective off-pump coronary bypass operation is possible with a low conversion rate and without an apparent increase in morbidity or change in technique. Whereas short-term safety and efficacy seem certain, studies of long-term outcome are necessary before the eventual role of off-pump coronary bypass in myocardial revascularization can be defined. PMID- 11899184 TI - Morbidity after procurement of radial arteries in diabetic patients and the elderly undergoing coronary revascularization. AB - BACKGROUND: The use of radial arteries for coronary revascularization is increasing. There remain concerns regarding alteration of upper extremity function after radial artery procurement. This study evaluates the functional morbidity in higher risk patients. METHODS: Between April 1997 and September 1999, 374 patients underwent unilateral or bilateral radial artery procurement. A questionnaire was used to evaluate symptoms related to motor and sensory function and changes in appearance after radial artery harvest. RESULTS: Two hundred eighty-nine patients were successfully interviewed. The average age was 63 years. Median follow-up was 9.5 months (range, 2 to 23 months). No patient suffered limb loss. Altered gross and fine motor function, residual pain, paresthesias, numbness, pallor, swelling, and altered temperature sensation were compared among diabetic patients, patients older than 70 years, and patients without these characteristics. CONCLUSIONS: Radial artery procurement for elective coronary revascularization can be done with minimal serious morbidity in higher risk patients. The most common symptoms were numbness and paresthesia. Despite the finding of greater residual pain in diabetic patients, we do not believe the use of radial artery conduits is contraindicated in these patients. PMID- 11899186 TI - Robotically assisted versus conventional freehand technique during beating heart anastomoses of left internal thoracic artery to left anterior descending artery. AB - BACKGROUND: Although robotically assisted coronary arterial anastomoses are being performed clinically, the short-term and long-term quality and integrity of the left internal thoracic artery (LITA) to left anterior descending artery (LAD) anastomosis remains unknown. The goal of this study was to perform a histologic and angiographic assessment of porcine beating heart LITA to LAD anastomoses using either robotic assistance or a conventional freehand technique. METHODS: Twelve pigs underwent beating heart LITA to LAD anastomoses using either the robotically assisted (n = 6) or conventional freehand techniques (n = 6). Quantitative histologic analysis was performed in all animals in order to determine the degree of vascular wall damage. Selective coronary arteriography was performed in all animals immediately after the procedure in order to evaluate anastomotic patency. The unpaired Student's t test was used for all comparisons between groups. RESULTS: There were no differences in vascular wall damage between the robotically assisted and freehand techniques. Postoperative angiography revealed no stenoses in either group. CONCLUSIONS: Use of the robotically assisted technique for creation of a LITA to LAD anastomosis was not associated with increased histologic damage when compared with the freehand technique in a beating heart porcine model. Furthermore, there was no difference between the two techniques in postoperative patency rate. These results support further clinical investigation of robotically assisted coronary bypass surgery. PMID- 11899185 TI - The importance of intraoperative angiographic findings for predicting long-term patency in coronary artery bypass operations. AB - BACKGROUND: The quality of anastomosis is the cornerstone in coronary artery bypass operations. Intraoperative coronary angiography confirms graft patency with the possibility to revise graft failure. The aim of this study was to describe the lesions found at "on-table" angiography, and to evaluate the significance of these immediate angiographic findings for the long-term patency. METHODS: A total of 57 grafts (42 left internal mammary artery grafts and 15 saphenous vein grafts) in 45 patients who underwent off-pump coronary artery bypass operations were included. On-table angiography was carried out with fixed angiographic equipment installed in the operating room. Follow-up angiographies were performed at 3 months and at 12 months. RESULTS: The most frequent finding in an on-table angiogram was spasm, which was not present at follow-up. Out of nine kinks, only one developed into a significant stenosis at follow-up. Of 44 grafts that were normal on-table, 37 (84%) were normal at the follow-up. Of 11 grafts with significant lesions on-table, eight (73%) were normal at the follow up. Five percent of the grafts were revised because of the on-table angiography. CONCLUSIONS: On-table angiograms can be occasionally difficult to interpret because not all findings are important for later patency. Optimal results on table predict good long-term results with a negative predictive value of 0.84, whereas significant lesions on-table have less impact on the follow-up results because the positive predictive value was only 0.38. PMID- 11899187 TI - An experimental model of saphenous vein-to-coronary artery anastomosis with the St. Jude Medical stainless steel connector. AB - BACKGROUND: A new stainless steel anastomosis device developed by St. Jude Medical Cardiovascular Group was studied in a canine model. METHODS: In 12 dogs, coronary saphenous vein grafts were made to the left anterior descending coronary artery and to the circumflex coronary artery; one anastomosis was completed with the St. Jude Medical stainless steel connector device, and the other with conventional suturing. A 30-day coronary angiogram was performed in surviving animals, and, after sacrifice, anastomoses were measured, examined grossly, and submitted for histologic study. RESULTS: All 12 animals survived the procedure, and 9 survived to sacrifice at 30 days. Comparing the connector grafts and sutured grafts, no significant differences were found between vessel diameters, intraoperative graft flows, graft patency, and histology. The average loading time for the connector was 8.5 minutes (range 4 to 16 minutes). Mean time for the 12 connector anastomoses was 3 minutes (range 2 to 5 minutes) compared with 8.4 minutes for suture (range 4 to 13 minutes). CONCLUSIONS: The side-to-side stainless steel connector anastomotic device produces a secure anastomosis with minimal variability; compared with suture methods, it is expeditious and has comparable 30-day histology and angiographic results. It promises to be an important addition to the surgical armamentarium for the treatment of coronary artery disease. PMID- 11899188 TI - Cardioplegic arrest with L-arginine improves myocardial protection: results of a prospective randomized clinical trial. AB - BACKGROUND: Blood cardioplegic arrest remains the method of choice for myocardial protection. L-arginine has been suggested to improve protection through an increase in nitric oxide production. METHODS: A prospective, randomized, double blinded clinical trial comparing standard blood cardioplegic solution to L arginine-enriched solution (7.5 g/500 mL) enrolled 200 patients undergoing coronary artery bypass grafting. Clinical data and biochemical markers of ischemia were recorded. Warm blood cardioplegia (33 degrees C) was administered in 74% of patients and cold blood (20 degrees C) was used in 26% of patients. Both groups averaged three grafts per patient. RESULTS: There were two (2%) deaths in both groups. There were four (4%) myocardial infarctions (MI) in the control group and six (6%) infarctions in the L-arginine group (p = 0.5). For the 190 patients without MI, serum levels of troponin T averaged 0.40+/-0.43, 0.38+/ 0.42, and 0.39+/-0.50 microg/L in control patients compared with 0.28+/-0.22, 0.24+/-0.18, and 0.27+/-0.20 microg/L in L-arginine patients, respectively, 12, 24 and 48 hours after coronary artery bypass grafting (p = 0.03). The cardiac index averaged 2.7+/-0.8 L x min(-1) x m(-2) in control patients and 2.9+/-0.7 L x min(-1) x m(-2) in arginine patients immediately after surgery (p = 0.09). Intensive care unit and hospital length of stay averaged 3.5+/-5 days and 7.3+/-6 days in control patients compared with 2.5+/-3 days and 6.1+/-4 days in arginine patients (p = 0.09). CONCLUSIONS: L-arginine-supplemented blood cardioplegic solution is associated with reduced release of biochemical markers of myocardial damage, suggesting improved myocardial protection. PMID- 11899190 TI - Functional recovery of the native heart after cardiomyoplasty in sheep with heart failure: passive and dynamic effects of volume loading. AB - BACKGROUND: Dynamic cardiomyoplasty (d-CMP) encourages reverse remodeling and improved contractility and stroke work (SW) efficiency of the failing native heart. This contrasts with passive cardiomyoplasty (p-CMP), which provides "passive girdling." To further evaluate pump recovery we assessed native left ventricular performance (without assist) 6 months after dynamic and passive CMP in sheep with heart failure with acute volume loading. METHODS: Heart failure (left ventricular ejection fraction 26%+/-8%) induced by coronary microembolization was followed by CMP in 11 sheep. After 8 weeks of muscle "training," paced cardiac assist was undertaken in the d-CMP group (n = 6). Five sheep with heart failure served as controls. Six months later the pressure-volume relationship was derived before and after volume loading by colloid solution. Latissimus dorsi muscle pacing was previously ceased in the d-CMP group. RESULTS: Volume loading increased left ventricular end-diastolic volume and pressure in all groups. After volume loading in d-CMP, the SW and pressure-volume area were increased, and SW efficiency remained unchanged. In p-CMP neither variable changed, whereas in control heart failure SW efficiency decreased due to a rise in pressure-volume area with stable SW. CONCLUSIONS: Based on response to volume loading, the failing native heart after 6 months of d-CMP showed functional recovery from "active girdling," whereas p-CMP prevented functional deterioration through passive girdling. The failing control heart progressively deteriorated. PMID- 11899189 TI - Is there a place for preconditioning during cardiac operations in humans? AB - BACKGROUND: Activation of the kinase cascade (protein kinase C (PKC), tyrosine kinase (TK), and mitogen-activated protein kinase (MAPK) is a key feature of the transduction pathway, elicited by preconditioning signals and mediating their cardioprotective effects. We assessed whether such an activation occurred during cardiac operations and could thus represent a target for cardioprotective strategies. METHODS: A total of 20 patients undergoing coronary artery bypass grafting surgery were studied. During the first 10 minutes of cardiopulmonary bypass (CPB), 10 were treated with sevoflurane (2.5 minimum alveolar concentration), an inhalational anesthetic that mimics preconditioning through a similar activation of the kinase cascade. Ten case-matched patients undergoing 10 minutes of sevoflurane-free CPB served as controls. Right atrial biopsies were taken before and 10 minutes after CPB and were then processed for the measurement of PKC, TK, and p38 MAPK activities by enzyme assay techniques. Troponin I was also monitored over the first 2 postoperative days. RESULTS: Compared with pre CPB values, PKC and p38 MAPK activities (in nanomoles per milligram of protein per minute and arbitrary units, respectively) increased significantly and to the same extent in both groups: PKC, from 20.7+/-0.7 to 29.9+/-3.9 in controls (p = 0.037) and from 18.4+/-1.1 to 23.9+/-1.8 in sevoflurane (p = 0.016); p38 MAPK, from 88.6+/-8.5 to 312.9+/-66.2 in controls (p = 0.005) and from 114.6+/-14.7 to 213.4+/-51.8 in sevoflurane (p = 0.045). Conversely, sevoflurane triggered a significant increase in TK activity (from 68.5+/-1.4 to 83.7+/-2.9 picomoles per milligram of protein per minute p = 0.0015) which did not occur in controls (from 67.5+/-1.9 to 76.8+/-4.2 picomoles per milligram of protein per minute, p = 0.09). Likewise, the peak postoperative value of troponin I was not different between controls and sevoflurane-treated patients (3.4+/-0.6 vs 2.4+/-0.4, p = 0.21). CONCLUSIONS: Cardiopulmonary bypass triggers an activation of the kinase cascade that is mechanistically linked to opening of potassium channels. The direct opening of these channels by the anesthetic sevoflurane does not increase kinase activation further, nor does it improve markers of cell necrosis, thus suggesting that pharmacologically targeting potassium channels may overlap the preconditioning-like effects of CPB alone. PMID- 11899191 TI - Midterm results of surgical treatment of systemic ventricular outflow obstruction in Fontan patients. AB - BACKGROUND: Achieving unobstructed blood flow from the systemic ventricle to the aorta is important during the Fontan procedure for complex cyanotic congenital heart disease when there is systemic ventricular outflow obstruction (SVOO). Because SVOO can progress after the Fontan procedure if there is morphologic obstruction, we have adopted a policy of relieving obstructions to systemic blood flow. METHODS: Twenty-five patients were treated by the Fontan procedure with SVOO. Twenty-one patients had undergone prior pulmonary artery banding and 10 patients had undergone prior arch repair. Systemic ventricular outflow obstruction progressed in 5 patients after the Fontan procedure. Main diagnosis was single ventricle in 12, tricuspid atresia in 5, transposition of the great arteries in 4, double-outlet right ventricle in 3, and common atrioventricular canal in 1. Mean age at operation was 6.5 years (range 1 to 15 years) and the average preoperative pressure gradient across the ascending aorta and systemic ventricle was 29 mm Hg (range 0 to 100 mm Hg). The Damus-Kaye-Stansel procedure was performed in 18 patients (double-barrel anastomosis in 13, end to side anastomosis in 5), and subaortic resection or ventricular septal defect or bulboventricular foramen enlargement was performed in 7. Double-barrel anastomosis has been our first choice since 1994, if the pulmonary valve is intact. Follow-up has ranged from 4 months to 14 years (average 5.0 years). Twenty-three of the 25 patients have undergone recatheterization (average 21.4 months later). RESULTS: No early deaths were found; one late death was reported of a patient with single right ventricle (4.0%). The postoperative average pressure gradient was 1.1 mm Hg (0 to 10 mm Hg), and the average right atrial pressure was 14 mm Hg (9 to 20 mm Hg). In all patients who underwent ventricular septal defect or bulboventricular foramen enlargement, regular sinus rhythm was maintained postoperatively. Regarding the Damus-Kaye-Stansel procedure, there was minimal progression of semilunar valve insufficiency except in 1 patient who underwent end-to-side anastomosis with moderate pulmonary regurgitation postoperatively. CONCLUSIONS: The midterm results of the Fontan procedure with SVOO have been satisfactory. Because SVOO might progress after the Fontan procedure if there is morphologic obstruction, an appropriate strategy to relieve obstruction to systemic blood flow should therefore be performed concomitantly with the Fontan procedure. PMID- 11899192 TI - Effects of dilutional and modified ultrafiltration in plasma endothelin-1 and pulmonary vascular resistance after the Fontan procedure. AB - BACKGROUND: Pulmonary vascular resistance (PVR) is closely related with patients' hemodynamics after the Fontan procedure and endothelin-1 (ET-1) may play an important role in pulmonary circulation. Modified ultrafiltration (MUF) is known to remove inflammatory mediators after cardiopulmonary bypass (CPB) surgery. The time courses of plasma ET-1 and PVR were examined before and after the Fontan procedure with MUF. METHODS: Twenty-two patients who underwent the Fontan procedure were divided into two groups: a dilutional ultrafiltration/modified ultrafiltration (DUF/MUF) group (n =11) and a control group (n = 11). Conventional ultrafiltration was performed during CPB in the control group. DUF was performed semicontinuously during CPB and MUF was continued until 15 to 20 minutes after the CPB with polyacrylonitonile membrane in the DUF/ MUF group. The plasma ET-1 concentration was measured before and after CPB, after MUF in the DUF/MUF group, and 6 and 24 hours after CPB. PVR was calculated simultaneously using a thermodilutional catheter. RESULTS: Plasma ET-1 levels increased significantly after CPB in the control group but they did not increase immediately after CPB in the DUF/MUF group. Similarly, PVR increased significantly after CPB in the control group but it did not increase after CPB in the DUF/MUF group and remained low at 6 and 24 hours after CPB. CONCLUSIONS: DUF and MUF suppress the increase in the plasma ET-1 concentration that occurs immediately after the completion of the Fontan procedure and may be an effective intervention for maintaining low PVR after the procedure. PMID- 11899193 TI - Extending the usable size range of homografts in the pulmonary circulation: outcome of bicuspid homografts. AB - BACKGROUND: Small-sized homografts are often not available, making the use of surgically reduced cryopreserved homograft conduits appealing. METHODS: From January 1993 to January 2000, 21 large homografts were size-reduced by excising one leaflet and were implanted in the pulmonary circulation. Valve function was compared with 21 children-matched for weight, homograft size, and year of operation-who received a standard homograft. RESULTS: Implanted homograft size and patient weight were equivalent in both the bicuspid and standard groups. Median (range) in-hospital peak instantaneous echocardiographic gradient across the valve was 0 mm Hg (0 to 19) in the bicuspid group versus 0 mm Hg (0 to 17) in the standard group (p = 0.65). Median (range) in-hospital pulmonary insufficiency (scale of 0 to 4) was 2 (0 to 3) in the bicuspid group versus 1.5 (0 to 3) in the standard group (n = 10, p = 0.34). At a follow-up of 54+/-29 months there was no significant difference in conduit reinterventions between the groups. CONCLUSIONS: Surgical creation of a bicuspid valve in the pulmonary circulation results in a functionally equivalent conduit compared with standard homograft as measured by early and midterm valve function. PMID- 11899194 TI - PTFE monocusp valve reconstruction of the right ventricular outflow tract. AB - BACKGROUND: Transannular patching of right ventricular outflow tract obstructive (RVOTO) defects results in pulmonary insufficiency (PI). Biologic monocusp valves (MO) can prevent acute PI but are prone to early degeneration and progressive regurgitation. Polytetrafluoroethylene (PTFE, 0.1 mm) MO leaflets demonstrated favorable characteristics in animal studies, and the technique was applied to a variety of RVOTO anomalies. METHODS: From June 1990 through June 1999, 158 patients underwent either PTFE MO RVOT reconstruction (n = 115 patients; 120 implants) or nonvalved transannular repair (TA) repairs (n = 43 patients; 5 subsequent MO implants) at our institution. Standard MO construction techniques and TA repairs were utilized. Intraoperative, postoperative, and echocardiographic data with a mean interval of 2.6 years (range 6 months to 8 years) were used in retrospective fashion to compare clinical outcomes. In addition, PTFE monocusp valves beyond 6 months postimplant underwent echocardiographic analysis of MO function and durability. RESULTS: There were 4 early (MO-3, TA-1) and no late deaths. Overall, perioperative complications were not significantly different between MO and TA groups, nor were total hospitalization days (9.1 versus 10.7, p = 0.24). However, a significant difference in intensive care unit (ICU) utilization (3.6 versus 5.8 days, p = 0.03) favored MO patients. Patients with tetralogy of Fallot (TOF) and ventricular septal defect/pulmonary atresia (VSD/PA) undergoing the MO implant demonstrated a trend toward improved survival (p = 0.08) when compared to TA repairs. Intraoperative PI was graded mild in the MO group and moderate-severe in the TA group (p = 0.003). Progressive MO regurgitation occurred (mild-moderate) but remained significantly less than the transannular patch repairs (p < 0.05). CONCLUSIONS: Utilization of a PTFE MO valve prevents short-term and significantly reduces midterm PI. It is inexpensive, easy to construct, and demonstrates no evidence of stenosis, calcification, or embolization. Despite longer cardiopulmonary bypass and ischemic times, it reduces ICU stay and, in both TOF and VSD/PA patients, decreases operative morbidity and mortality. PMID- 11899195 TI - Bronchial compression by posteriorly displaced ascending aorta in patients with congenital heart disease. AB - BACKGROUND: We encountered several patients with posteriorly displaced ascending aorta and bronchial compression associated with congenital heart disease. We describe the helical computed tomography (CT) findings and explore the mechanism of airway compression. METHODS: We retrospectively reviewed the clinical data and CT findings of 8 patients with posterior displacement of the ascending aorta. The bronchial stenosis was quantified on reformatted images perpendicular to the main stem bronchi. On an axial image at the level of main bronchi, we measured depth of retrosternal space, interaortic distance, and aorto-spinal distance. To compare with control, we measured the same variables in 10 control patients. RESULTS: In 7 patients, the main bronchus on the side of the aortic arch was squeezed between the ascending and descending aorta and showed slit-like stenosis. The right pulmonary artery was elongated around the ascending aorta in 5 patients and showed slit-like stenosis in 3. Patients with posterior displacement had significantly larger retrosternal space, smaller interaortic distance, and smaller aorto-spinal distance than did the control group. Aortopexy was undertaken in 3 patients. Follow-up computed tomograms of 2 patients showed improvement. CONCLUSIONS: The posteriorly displaced ascending aorta may compress the main bronchus on the side of the aortic arch and right pulmonary artery against the descending aorta or spine. Even if the bronchial compression is mild with tolerable airway symptoms, these patients must be closely observed. When airway symptoms are severe, aortopexy should be considered. PMID- 11899196 TI - Current results with intraaortic balloon pumping in infants and children. AB - BACKGROUND: Intraaortic balloon pumping (IABP) is useful for support in patients with moderate left ventricular dysfunction. IABP is usually timed with the R wave of the electrocardiogram. We have utilized M-mode echocardiography timed IABP in children with left-side heart failure since 1994. Electrocardiogram timing seems inappropriate for children, who have much higher heart rates. We describe our experience with children who underwent IABP therapy before and after 1994, when echocardiographic timing was instituted. METHODS: We reviewed records of 29 children who underwent IABP for all indications at Primary Children's Medical Center since 1988. RESULTS: Overall survival was 62.1% (18 of 29) in this series. Survival was similar for infants (odds ratio = 2.0, 95% confidence interval = 0.29 to 14.31, p = 0.43) and older children. Survival was similar in the echocardiography era when compared with the electrocardiogram era (odds ratio = 2.4, 95% confidence interval = 0.56 to 10.4, p = 0.44). CONCLUSIONS: IABP is a useful means of support in children with left ventricular dysfunction. M-mode echocardiography is effective in triggering IABP. The sample size in this study is too small to detect a mortality rate difference. PMID- 11899197 TI - Thoracic duct tributaries from intrathoracic organs. AB - BACKGROUND: The thoracic duct (TD) is the main collecting vessel of the lymphatic system. Little is known about the intrathoracic tributaries of the TD, which are named intercostal, mediastinal, and bronchomediastinal trunks. The purpose of the study was to identify the lymphatic tributaries from intrathoracic organs to the thoracic duct. METHODS: The study was performed on 530 adult cadavers. The lymphatics of different organs were catheterized and injected with a dye: lungs (n = 360), heart (n = 90), esophagus (n = 50), and diaphragm (n = 30). The lymphatic tributaries draining the lymph from these organs to the thoracic duct were dissected along their course to the thoracic duct and classified. RESULTS: The TD tributaries were observed in 147 cases: right lung (n = 46), left lung (n = 69), heart (n = 8), esophagus (n = 13), and diaphragm (n = 11). Connections with the TD were observed at its origin (n = 13), within the mediastinum (n = 87), and at the level of the TD arch (n = 47). Tributaries from the lung issued from lower paratracheal nodes 4 R (n = 14) and 4 L (n = 31), subaortic 5 (n = 4), subcarinal 7 (n = 18), pulmonary ligament 9 (n = 7), upper tracheal 2 L (n = 28), paraortic 6 (n = 11), and celiac nodes (n = 2). Tributaries from the heart connected with the TD in the mediastinum in 1 case (4 L) and with the TD arch in 7 cases. Tributaries from the esophagus connected with the thoracic duct within the mediastinum in 13 cases; anodal routes were frequent (n = 5). The TD tributaries from the diaphragm were observed in 11 cases, always connecting with the TD at its origin. CONCLUSIONS: Injection of intrathoracic organs permits visualization of TD tributaries. These tributaries appear located at unchanging levels. Lymph of intrathoracic organs may thus drain into the general circulation through the TD. The tributaries may represent a potential route for tumor cells dissemination. When incompetent, due to valve insufficiency, they permit chylous lymph to backflow into the intrathoracic lymph nodes. Injury at this level may lead to intrathoracic chylous effusions. PMID- 11899198 TI - A prospective trial of systematic nodal dissection for lung cancer by video assisted thoracic surgery: can it be perfect? AB - BACKGROUND: There have been no reports evaluating the completeness of systematic nodal dissection with video-assisted thoracic surgery (VATS). In order to elucidate the completeness of the dissection, we have conducted a prospective trial with patients having primary lung cancer. METHODS: Patients with clinical stage I lung cancer were the candidates for this study. Thoracotomy was performed with a small skin incision of 7 cm to 8 cm in length. Through these small wounds and two trocars, pulmonary resection was performed and then hilar and mediastinal lymph nodes were dissected. After that, a standard thoracotomy was carried out by another surgeon to complete systematic nodal dissection. RESULTS: Video-assisted thoracic surgery lobectomy with lymph node dissection was accomplished in 17 right lung cancer patients and 12 left lung cancer patients. On the right side, the average numbers of resected lymph nodes by VATS and remnant lymph nodes were 40.3 and 1.2, respectively. The average weights of dissected tissues by VATS and remnant tissues were 10.0 g and 0.2 g, respectively. On the left side, there were 37.1 and 1.2 lymph nodes and 8.3 g and 0.2 g of weight of dissected tissues. No nodal involvement was observed in the remnant lymph nodes. CONCLUSIONS: The lymph node dissection with VATS was technically feasible and the remnant ("missed" by VATS) lymph nodes and tissues were 2% to 3%, which seems acceptable for clinical stage I lung cancer. PMID- 11899199 TI - Ganciclovir prodrug therapy is effective in a murine xenotransplant model of human lung cancer. AB - BACKGROUND: Therapy failures have been reported for retroviral gene transfer of herpes simplex virus thymidine kinase (HSV-TK) gene followed by systemic ganciclovir application in human lung cancer. Use of the HSV-TK mutant TK30 in combination with a VSV-G pseudotyped retroviral vector was found to enhance the efficacy of prodrug therapy. The present study evaluated this therapeutic strategy in human non-small cell lung cancer cell lines in a preclinical murine xenotransplant model. METHODS: Intrathoracally induced by HSV-TK30 transduced non small cell lung cancer cell lines Colo 699 (adenocarcinoma) and KNS 62 (squamous cell carcinoma) were monitored for local tumor growth, survival, and metastases. So-called bystander effects were investigated in tumors consisting of as little as 25% TK30 transfected cells and by analysis of gap junctional protein connexin 43 expression. RESULTS: Survival was significantly prolonged, and tumor growth and pleural metastases were reduced in HSV-TK30-positive tumors of both cell lines. A significant therapeutic effect in bystander experiments was observed in squamous cell carcinoma. This was correlated with higher expression of connexin 43. CONCLUSIONS: Delivery of HSV-TK30 in a VSV-G pseudotyped retroviral vector and subsequent ganciclovir application provided therapeutic efficacy. Despite of low transduction rates achievable in gene transfer in situ, prodrug therapy appears to be feasible in tumor cells with efficient bystander effects. PMID- 11899200 TI - Benign broncho-esophageal fistula in the adult. AB - BACKGROUND: Benign broncho-esophageal fistula (BEF) in the adult is rare, and occurs as a complication of inflammatory disorders, foreign body ingestion, or congenital anomalies. Nonspecific symptoms lead to a delay in diagnosis. METHODS: The charts of 13 patients from 1960 to 2001 at the Massachusetts General Hospital were retrospectively reviewed. RESULTS: Nine patients had chronic cough, which worsened upon ingestion. Four patients developed BEF after prior thoracic surgery, and 3 after histoplasmosis. Silicosis, foreign body ingestion, lye ingestion, bronchogenic cyst, esophageal diverticulum, and a congenital anomaly caused BEF in 1 patient each. Barium swallow was the most useful diagnostic test. Fistulas most often arose from the right bronchial tree and communicated with the distal esophagus. Management involved excision of the tract, primary closure of the bronchus and esophagus, and interposition of vascularized tissue. There was one perioperative failure, but no long-term recurrences after successful surgical closure. CONCLUSIONS: The majority of benign BEF in adults are acquired, and result from mediastinal inflammation. Accurate recognition and surgical closure prevents persistent uncontrolled aspiration and pulmonary sepsis. PMID- 11899201 TI - Clinical impact of endoscopic ultrasound-guided fine needle aspiration of celiac axis lymph nodes (M1a disease) in esophageal cancer. AB - BACKGROUND: The purpose of this study was to determine how endoscopic ultrasound (EUS)-guided fine needle aspiration (FNA) with a histology confirmed biopsy protocol impacted on staging and managing esophageal carcinoma in terms of resectability and neoadjuvant therapy (chemotherapy and radiation therapy). METHODS: The records of 40 consecutive patients diagnosed with esophageal cancer referred for EUS staging were reviewed. Computed tomography (CT) scan then EUS imaging and EUS-guided FNA staging, including involvement of celiac node (M1a stage), surgical pathology, and subsequent treatment were correlated. Through-the scope balloons were used for dilatation when needed to examine the celiac nodes. RESULTS: All 40 patients followed the protocol and were successfully imaged by EUS. Sixteen of the 40 required esophageal dilatation using the through-the-scope balloon. No complications were observed from esophageal dilatation for EUS. Twenty-three (58%) met the criteria for EUS-guided FNA biopsy from a total of 40 EUS imaging procedures. Twenty (87%) of the 23 EUS-guided FNA were directed toward the celiac nodes; 18 (90%) of the 20 were positive for malignancy and were treated by chemoradiation therapy and 2 (10%) FNA were negative for malignancy and were treated by surgical resection. The CT scan was able to detect only 6 (30%) of 20 cases of suspicious celiac lymph nodes, of which 5 (83%) were positive for malignancy by FNA. CONCLUSIONS: EUS-guided FNA of celiac nodes (20 patients) directed management in all patients biopsied. EUS-guided FNA is superior to CT scan for diagnosing M1a disease. Protocol-directed EUS-guided FNA is a pivotal study when used in conjunction with stage-oriented treatment protocols for esophageal carcinoma. PMID- 11899202 TI - Pulmonary complications after esophagectomy. AB - BACKGROUND: Pulmonary complications are common in patients who have undergone esophagectomy. There are no good predictive variables for these complications. In addition, the role that preoperative treatment with chemotherapy and radiation may play in postoperative complications remains unclear. METHODS: We performed a retrospective review of all patients who underwent esophagectomy by a single surgeon at our institution over a 6-year period. Data were analyzed for a correlation between patient risk factors and pulmonary complications, including mortality, prolonged mechanical ventilation, and hospital length of stay. RESULTS: Complete data were available on 61 patients. Nearly all patients had some pulmonary abnormality (eg, pleural effusion), although most of these were clinically insignificant. Pneumonia was the most common clinically important complication, and 19.7% of patients required prolonged ventilatory support. Significant risk factors identified included impaired pulmonary function, especially for patients with forced expiratory volume in 1 second (FEV1) less than 65% of predicted, preoperative chemoradiotherapy, and age. CONCLUSIONS: Impaired lung function is a significant risk factor for pulmonary complications after esophagectomy. Patients with FEV1 less than 65% of predicted appear to be at greatest risk. There also seems to be an associated risk of preoperative chemoradiotherapy for pulmonary complications after esophagectomy. PMID- 11899203 TI - Clinical significance of telomerase activity in peripheral blood of patients with esophageal squamous cell carcinoma. AB - BACKGROUND: The presence of tumor cells in the blood stream is considered evidence of a high risk of distant organ metastasis. We examined the usefulness of telomerase activity in peripheral blood polymorphonuclear cells as an indicator of distant metastasis in patients with esophageal squamous cell carcinoma. METHODS: Telomerase activity was measured in the peripheral blood mononuclear cell and polymorphonuclear cell fractions obtained from blood samples of healthy volunteers mixed with squamous cell carcinoma cell lines, and cell distribution was analyzed by flow cytometry. Then telomerase activity of forty two polymorphonuclear cell fractions obtained from esophageal squamous cell carcinoma patients was measured. RESULTS: Telomerase activity was detected in polymorphonuclear cell fractions and cell distribution analysis revealed the presence of esophageal squamous cell carcinoma cells. Organ metastasis was detected in 7 (78%) of the 9 patients with telomerase-positive polymorphonuclear cell fractions as opposed to only five (15%) of the 33 with telomerase-negative cases, and there was a significant positive correlation between telomerase activity and organ metastasis (p < 0.0008). CONCLUSIONS: Measurement of telomerase activity in the polymorphonuclear cell fractions is useful for identifying a high risk group for distant organ metastasis in patients with esophageal squamous cell carcinoma. PMID- 11899204 TI - Expression status of E-cadherin and alpha-, beta-, and gamma-catenins in thymoma. AB - BACKGROUND: A loss or dysfunction of E-cadherin or catenins, which maintain tissue integrity, is associated with an invasive phenotype of various solid tumors. Therefore, we analyzed the expression of E-cadherin and alpha-catenin, beta-catenin, and gamma-catenin in thymoma tissue specimens to investigate its clinical significance. METHODS: The expressions of E-cadherin and alpha-catenin, beta-catenin, and gamma-catenin in thymoma tissues were evaluated in 21 patients, including 9 epithelial predominant type, 5 lymphocytic predominant type, and 7 mixed type patients based on an immunohistochemical analysis using monoclonal antibodies, and the relationship between the expression status and clinicopathologic features was investigated. RESULTS: Reduced expressions were observed in 11 patients (52%) for E-cadherin, 10 (45%) for alpha-catenin, 6 (27%) for beta-catenin, and 10 (45%) for gamma-catenin. Such an expression status (reduced or preserved) of the molecules closely correlated with each other. The expression of E-cadherin was well preserved in 5 of 5 patients with lymphocyte predominant type whereas E-cadherin was reduced in 11 of 17 patients with other histologic subtypes. All of the 9 cortex type thymomas (B1 to 3) showed preserved expression of beta-catenin. There was no significant relationship among the expressions of the molecules and the Masaoka stage classification (I versus others). CONCLUSIONS: The status of expressions for these molecules may affect the degree of lymphoid infiltration while not affecting the degree of invasiveness in thymoma. PMID- 11899205 TI - Experience with Ultraflex expandable metallic stents in the management of endobronchial pathology. AB - BACKGROUND: Experience with Ultraflex expandable metallic stents (Micro-invasive, Boston Scientific, Watertown, MA) in the management of endobronchial pathologies leading to airway compromise is reported. METHODS: Between January 1999 and August 2000, twenty-eight expandable metallic stents were inserted into 25 patients (7 men and 18 women; median age, 65 years) who presented with respiratory distress. Each patient had comorbid medical conditions or end-stage malignancy that precluded formal surgical repair. Seventeen patients had intrinsic airway obstruction, 5 had extrinsic compression, 2 had a tracheal tear, and 1 had a tracheoesophageal fistula. Stents were inserted through a bronchoscope under direct vision. Eighteen patients received tracheal stents alone (1 of these patients received two tracheal stents), and 5 patients received bronchial stents only. Two patients received a tracheal and a bronchial stent. Twenty-one stents were covered and seven were uncovered. RESULTS: All patients had successful stents with restoration of airway patency and closure of tracheal defects. One patient developed a respiratory infection early after the operation. Follow-up bronchoscopy confirmed satisfactory stent position in each patient. Late complications included sputum retention, halitosis, and granulation tissue formation. CONCLUSIONS: Ultraflex expandable metallic stents should be considered in the management of airway compromise in selected patients for whom formal surgical repair is inappropriate or contraindicated. PMID- 11899207 TI - Preoperative fenestration for type A acute aortic dissection with mesenteric malperfusion. AB - We report a case of a 71-year-old man presenting with acute type A aortic dissection and mesenteric ischemia due to extension of the intimal flap to the mesenteric artery. Because of the severity of the abdominal symptoms, surgical correction of the ascending aorta was delayed. Preoperative percutaneous fenestration was performed successfully, allowing ascending aortic replacement 6 days later. Transverse colon stenosis secondary to preoperative ischemia occurred in the postoperative course. The patient was discharged from hospital with normal intestinal transit 72 days later. PMID- 11899206 TI - Endobronchial neodymium:yttrium-aluminum garnet laser for noninvasive closure of small proximal bronchopleural fistula after lung resection. AB - BACKGROUND: Bronchopleural fistula (BPF) is a serious complication of lung resection. The management of persistent BPF is one of the most complex challenges encountered by thoracic surgeons. METHODS: We used neodymium:yttrium-aluminum garnet (Nd:YAG) laser in 8 patients with BPF who were treated at our hospital, between January 1991 and December 1997. Through the flexible fiberoptic bronchoscope, Nd:YAG laser beam was directed to the bronchial mucosa surrounding the BPF. One-half-second energy pulses of 8 to 20 W were used. Close follow-up of successful patients showed complete closure of the BPF without further treatment. RESULTS: The procedure was successful in 4 of 5 patients who had no infection or tumor at the bronchial stump. However, the procedure failed in 3 other patients, who had residual tumor or infection by aspergillus at the stump. CONCLUSIONS: Closure of small BPF by laser seems to be due to edema and to an inflammatory reaction of the bronchial mucosa surrounding the BPF. If the diagnosis of small proximal BPF is made in the absence of tumor or infection, Nd:YAG laser offers an option for endobronchial treatment of small (<2 mm) BPF. If this technique is successful, it avoids the morbidity associated with more invasive surgical procedures. PMID- 11899208 TI - Traumatic aortic transection: evidence for the osseous pinch mechanism. AB - Acute traumatic transection of the thoracic aorta is most commonly seen in vehicular trauma and is generally accepted to be due to differential deceleration. A second mechanism is proposed for this injury and that is the osseous pinch mechanism. We report a case where aortic transection occurred due to a crush injury and supports the latter mechanism. PMID- 11899209 TI - Successful stent-grafting for perforation of the thoracic aorta by an intraaortic balloon pump. AB - Vascular complications associated with intraaortic balloon pump placement are quite common and predominantly related to femoral or iliac damage. Iatrogenic injury of the thoracic aorta is less usual and often fatal. Surgery for the lesions of the descending thoracic aorta still has a relatively high morbidity and mortality. Endovascular covered stentgraft prostheses have become a less invasive therapeutic approach to lesions of the thoracic aorta, especially in patients with high surgical risk. We describe a case of perforation of the thoracic aorta caused by an intraaortic balloon pump. The injury was confirmed by aortography and successfully repaired by implantation of an endovascular stent graft via the left common iliac artery. PMID- 11899210 TI - Intramural hematoma of the thoracic aorta in a woman with neurofibromatosis. AB - The natural history and preferred approach to patients with intramural hematoma of the aorta remains controversial. We describe herein the management of the case of a 45-year-old woman with acute onset of chest pain, intramural hematoma of the arch and proximal descending aorta, and left pleural effusion. In this particular case, a nonsurgical approach led to complete recovery of the patient and documented resolution of the hematoma. PMID- 11899212 TI - Ex-vivo mitral valve repair prior to orthotopic cardiac transplantation. AB - Mitral valve annuloplasty was performed prior to orthotopic cardiac transplantation in two donor hearts which were diagnosed with moderate to severe mitral regurgitation. The technical aspects are reviewed of ex-vivo mitral valve repair with concomitant heart transplantation. The recipients were classified as United Network for Organ Sharing (UNOS) I and both patients have had an excellent postoperative recovery. Over 2-year follow-up demonstrates normal mitral valve function without regurgitation. PMID- 11899211 TI - Biventricular pacing for weaning from extracorporeal circulation in heart failure. AB - Resynchronization of the intra- and interventricular conduction by biventricular pacing has been suggested in patients with end-stage heart failure. We present a case in which extracorporeal circulation could only be weaned after placement of an additional left ventricular pacing wire. Biventricular stimulation led to normal motion of the anterior wall and a previously bulging interventricular septum; this improved the hemodynamic situation significantly. PMID- 11899213 TI - Correction of traumatic tricuspid regurgitation using the double orifice technique. AB - We present a case of acute traumatic tricuspid regurgitation in a 39-year-old man who was involved in a motor vehicle accident. A large ecchymotic region over the anterior chest wall prompted evaluation by both transthoracic and transesophageal echocardiography which confirmed the valvular injury. At surgery, valvular incompetence was found to be the result of a flail anterior leaflet due to papillary muscle rupture. The valve was successfully repaired using a single stitch double orifice technique in combination with a ring annuloplasty. The valve remains competent 18 months after surgery. PMID- 11899214 TI - Replacement of a thrombosed valve after the Bentall procedure. AB - A 50-year-old man developed thrombosis in the valve of a Bjork-Shiley prosthesis that had been used for composite graft replacement of the aortic valve and ascending aorta 8 years previously. The thrombosed valve was removed, and because of the narrow aortic valve ring, it was replaced using patch enlargement of the aortic annulus without replacement of the conduit. PMID- 11899215 TI - Tetralogy of Fallot with aortic valvular stenosis: surgical correction in one case. AB - We report a case of an infant presenting with the rare association of tetralogy of Fallot, hypoplasia of the pulmonary arteries, and stenotic bicuspid aortic valve. Surgical correction, performed at 16 months of age, included aortic valvular commissurotomy, opening the right ventricular outflow tract (transannular patch), and ventricular septal defect closure. The postoperative course was favorable, and the child was discharged from the hospital. Three months after the procedure, the patient is in excellent condition, without cardiac medication. PMID- 11899217 TI - Three-decade follow-up in pulmonary artery ectasia: risk assessment strategy. AB - We present a 29-year follow-up in a patient with ectasia of the pulmonary trunk and mild valvular stenosis. There was no progression from the first presentation at the age of 15 months until the actual age of 30 years, the z-value of the diameter of the pulmonary trunk remaining almost constant (z-value 22). This indicates that pulmonary dilatation with normal pressure may be a benign disease. PMID- 11899216 TI - Ruptured right coronary artery aneurysm presenting as a myocardial mass. AB - Rupture of a coronary artery aneurysm may result in a contained hematoma and an intramyocardial mass. We present a case of a contained coronary artery aneurysm rupture presenting as a mass in the right ventricle. The mass was suspected in a preoperative transthoracic echocardiogram and identified at the time of elective coronary artery bypass surgery. We discuss the pathophysiology, clinical presentation, diagnostic features, and management of this uncommon clinical presentation. PMID- 11899218 TI - Successful arterial switch operation for post-Mustard pulmonary venous obstruction and secondary pulmonary hypertension. AB - A 16-year-old girl presented with dyspnea 15 years after the Mustard operation for transposition of the great arteries with intact ventricular septum. An echocardiogram revealed secondary pulmonary hypertension due to pulmonary venous obstruction. Cardiac catheterization showed the left (pulmonary) ventricular pressure was over the systemic level. We performed a successful one-stage switch conversion. The patient is doing well 1 year after the switch conversion. PMID- 11899219 TI - Treatment of persistent chylothorax after Norwood procedure with somatostatin. AB - A newborn who had undergone Norwood procedure for hypoplastic left heart syndrome developed a voluminous chylothorax that persisted despite weeks of prolonged complete bowel rest, total parenteral nutrition, and effective chest tube drainage. Chest tube output diminished immediately following initiation of intravenous somatostatin, allowing restoration of full enteral feeds and removal of chest tubes within 6 days. PMID- 11899220 TI - Sclerosing hemangioma with metastases to multiple nodal stations. AB - We present a case of a large pulmonary sclerosing hemangioma with metastases to multiple lymph nodal stations and suspected contralateral pulmonary metastasis. Four cases (including the present) have been reported to have lymph node metastasis, and all had large tumors exceeding 3.5 cm in diameter. Accordingly, resection of sclerosing hemangioma is advisable while the tumor is small. Even in cases with a large sclerosing hemangioma, lymph node metastasis may be uncommon. However lymph node dissection may be necessary to detect lymph node metastasis in selected cases. PMID- 11899221 TI - Preoperative embolization of a massive solitary fibrous tumor of the pleura. AB - We report the case of a 69-year-old woman with a massive fibrous tumor of the left hemithorax. Initial attempted removal by median sternotomy was abandoned due to the extremely vascular appearance of the tumor and the inability to gain safe control of the pedicle. Subsequent percutaneous embolization before removal through a left thoracotomy was successful, with little intraoperative blood loss. We recommend preoperative angiography for massive chest tumors that may be vascular and require piecemeal removal for total excision. PMID- 11899223 TI - Behavior of free jejunal flaps after early disruption of blood supply. AB - We describe three free jejunal flaps that lost their axial blood supply in the early postoperative period--two flaps on the 7th day and the third on the 17th day. At 7 days, reestablishment of axial blood flow was essential to flap survival, whereas after 17 days, vascularization from the recipient bed was adequate to maintain viability. Based on these observations, a conservative approach to flap salvage for cases with pedicle disruption at 17 days or later is recommended. PMID- 11899222 TI - Right aortic arch with left lung cancer: focusing on left recurrent laryngeal nerve. AB - We report a case of a 64-year-old Japanese man with an anomalous right aortic arch who had left lung cancer. We performed lobectomy and mediastinal lymphadenectomy, paying attention to the pathway of left recurrent laryngeal nerve. The left recurrent laryngeal nerve hooked around from the left dorsal to the right ventral part of the left ductus arteriosus, which connected the left pulmonary artery with the aortic diverticulum. PMID- 11899224 TI - Left atrial myxoma with tumor vascularity arising from the left and right coronary arteries. PMID- 11899225 TI - Single coronary artery-right ventricular fistula. PMID- 11899227 TI - Alternate technique for implantation of left ventricular assist system: left thoracotomy for reoperative cases. AB - Reoperation for Novacor left ventricular assist device placement after prior cardiac surgery is fraught with multiple technical challenges. We have found that a thoracotomy approach obviates these dangers very favorably. The technique is performed off bypass except for apical coring and apical connection. Novacor outflow is to the descending aorta. This approach has been found safe, quick, and effective. PMID- 11899226 TI - Modified extracardiac Fontan in a patient with separate hepatic venous drainage. AB - We describe an alternative technique to the extracardiac Fontan procedure in a patient with a univentricular heart, in which the inferior caval vein and the hepatic vein drained separately into the common atrium and the intraatrial orifice of the hepatic vein was adjacent to the opening of the left lower pulmonary vein. PMID- 11899228 TI - Linking CD4 gene expression and T cell development. AB - The control of CD4 gene expression is essential for proper T lymphocyte development. Signals transmitted from the T-cell antigen receptor (TCR) during the selection process are believed to be linked to the regulation of CD4 gene expression during specific stages of T cell development. Thus, the control CD4 gene expression is an ideal model system for studying the molecular mechanisms that drive T cell development. Here, I discuss the characterization of transcriptional control elements in the CD4 locus and the factors that mediate their function. The study of these elements has led to significant insights into the mechanisms in which the T lymphocyte develops it mature functional characteristics. PMID- 11899229 TI - Anti-CD45RB monoclonal antibody-mediated transplantation tolerance. AB - Currently, lifelong immunosuppression is required for organ transplant recipients. The majority of transplant recipients will eventually develop chronic rejection with resultant graft loss, despite treatment with powerful immunosuppressive agents. These agents are also associated with numerous toxicities including reduced immunity against infection and malignancy. Therefore, the central goal in transplant science is to devise tolerance strategies in an attempt to establish a state of prolonged non-reactivity against the allograft, accompanied with preservation of an intact immune system. Although predictable tolerance induction has been elusive, we found that short course of the novel immunomodulatory agent, anti-CD45RB monoclonal antibody, leads to indefinite acceptance of renal allografts in mice, and has been shown to markedly prolong allograft survival in primates. We review the current state of development of this antibody, and the progress made in defining its mechanism of action. PMID- 11899230 TI - How does HIV cause depletion of CD4 lymphocytes? A mechanism involving virus signaling through its cellular receptors. AB - HIV infection causes an acquired immunodeficiency, principally because of depletion of CD4 lymphocytes. The mechanism by which the virus depletes these cells, however, is not clearly understood. Since the virus predominantly infects CD4 lymphocytes in vivo, some have assumed that HIV replication directly kills the infected cells or that the anti-HIV immune response destroys them. However, a large number of studies do not support this concept. Rather, the data strongly indicate that CD4 lymphocyte depletion is by an indirect mechanism. Several theories on various direct and indirect mechanisms are reviewed. The most plausible mechanism, which is backed by in vivo data, involves the consequences of HIV contact with resting CD4 lymphocytes, which cannot support virus replication. HIV binding to, and signaling through, CD4 and chemokine receptor molecules on resting CD4 lymphocytes and other cell types [which extensively occurs as the rare, productively infected cells (ie: infected cells producing virus) migrate among other cells through the lymphoid tissues back into the blood] induces upregulation of L-selectin and Fas. When these resting, HIV signaled CD4 cells return to the blood, they home very rapidly back to peripheral lymph nodes and axial bone marrow, and their disappearance from the blood is likely due to their leaving the circulatory system. Approximately one-half of these cells that have been induced by HIV to home to lymph nodes are subsequently induced into apoptosis during the process of trans-endothelial migration when secondary signals are received through various homing receptors. These cells are not making HIV, which would explain the observation that CD4 cells not making HIV are the predominant cells dying in the lymph nodes of HIV+ subjects. These studies indicate that the principal mechanism of CD4 T-cell depletion by HIV is due to its use of CD4 as its primary receptor and the signaling induced through this receptor on nonpermissive (resting) T-lymphocytes. This unique mechanism of viral pathogenesis, if correct, leads to the possibility that HIV might not cause depletion of CD4 lymphocytes if it used some other receptor to infect CD4 lymphocytes. PMID- 11899231 TI - Molecular mechanisms of pulmonary fibrosis and current treatment. AB - Pulmonary fibrosis is a common response to various insults or injuries to the lung. Although there are various initiating factors or causes, the terminal stages are characterized by proliferation and progressive accumulation of connective tissue replacing normal functional parenchyma. The pathogenesis of pulmonary fibrosis includes endothelial and epithelial cell injury, production of inflammatory cells and their mediators, and fibroblast activation. Conventional therapy consisting of glucocorticoids or cytotoxic drugs is usually ineffective in preventing progression of the disease. Further understanding of the molecular mechanisms of endothelial and epithelial cell injury, inflammatory reaction, fibroblast proliferation, collagen deposition and lung repair, should lead to the development of effective treatments against pulmonary fibrosis. Accordingly, this review summarizes recent progress made in understanding the molecular mechanisms of pulmonary fibrosis. A detailed discussion is presented regarding each of the potential new therapies which have emerged from the animal models of pulmonary fibrosis and which have been developed through advances in cellular and molecular biology. PMID- 11899232 TI - Nucleic acid enzymes as a novel generation of anti-gene agents. AB - Recent molecular and cellular studies have highlighted the important role of some gene products in the cause and/or perpetuation of human pathological conditions including cancer and autoimmune diseases. The identification of such gene products has led to the development of new candidate therapies. The discovery of catalytic nucleic acid enzymes has provided researchers with a potentially important tool to block the expression of abnormal genes, provided that their sequences are known. The cleavage specificity of these compounds is determined by their hybridizing antisense arms, which anneal with the target mRNA in a complementary fashion. Nucleic acid enzymes can be delivered to cells either endogenously as gene encoding RNA enzymes (ribozymes) or exogenously as in vitro made agents. Given the progress reported during the last years, a wide range of molecular designs and chemical modifications can be introduced into these compounds, in particular the hammerhead type ribozyme. Here, we review the design, stability and the therapeutic application of these agents with the goals of illustrating relevant gene targets and signal pathways for molecular medicine. Relevant in vivo problems of the technology, mRNA repair by group I intron ribozymes and gene regulation by endogenous RNA will also be discussed. PMID- 11899233 TI - Receptors for unopsonized particles: the role of alveolar macrophage scavenger receptors. AB - The lung is constantly exposed to potentially pathogenic particles and microorganisms. Alveolar macrophage (AM) binding of inhaled environmental particles is a critical first step in phagocytosis and clearance, and must be accomplished without the benefit of opsonization by specific antibodies. Opsonin independent phagocytosis is initiated by direct recognition of phagocytic target. The identities of receptors on AMs that mediate unopsonized particle binding were, until recently, not known. Using flow cytometry, monoclonal antibody and expression cloning techniques we have found a major role for the scavenger receptor, MARCO in AM binding of particles and bacteria. In this review we will discuss the role of scavenger receptors in AM binding of unopsonized particles and the use of flow cytomety in analyzing AM-particle interaction. We will also discuss other non-scavenger receptors involved in opsonin-independent phagocytosis. PMID- 11899235 TI - Linking genomics to immunotherapy by reverse immunology--'immunomics' in the new millennium. AB - The disclosure of the human genome sequence and rapid advances in genomic expression profiling have revolutionized our knowledge about molecular changes in malignant diseases. Rapidly growing gene expression databases and improvements in bioinformatics tools set the stage for new approaches using large-scale molecular information to develop specific therapeutics in cancer. On one hand, the ability to detect clusters of genes differentially expressed in normal and malignant tissue may lead to widely applicable targeting of defined molecular structures. On the other hand, analyzing the 'molecular fingerprint' of an individual tumor raises the possibility of developing customized therapeutics. One approach to use the emerging new datasets for the development of novel therapeutics is to identify genes that are specifically expressed in tumors as targets for immune intervention. This review will focus on the process from in silico analysis of expression databases and screening of potential candidate genes by bioinformatics to the in vitro and in vivo analysis to determine the immunogenicity of candidate tumor antigens. Basic biological principles of 'reverse immunology' as well as technical advantages and difficulties will be addressed. PMID- 11899234 TI - Regulation of apoptosis and cell cycle activity in rheumatoid arthritis. AB - The regulation of proliferation and cell death is vital for homeostasis, but the mechanisms that coordinately balances these two events in rheumatoid arthritis (RA) remains largely unknown. In RA, the synovial lining increases through enhanced proliferation, migration, and/or decreased cell death. The aberrant decrease in apoptosis or increased cell cycle activity of fibroblast-like or macrophage-like synoviocytes is responsible for the synovial hyperplasia and contributes to the destruction of cartilage and bone. Recently, numerous molecules that modulate apoptosis and cell cycle have been implicated to play a role in RA. This review will describe the current understanding of the molecular mechanisms that govern apoptosis and cell cycle and their relationship to RA pathogenesis. PMID- 11899236 TI - The functional and clinical roles of osteopontin in cancer and metastasis. AB - Osteopontin (OPN) is a secreted and integrin-binding protein that has been implicated in a number of pathologies. In this review we will focus on the functional and clinical roles of OPN in cancer and metastasis, with a particular emphasis on breast cancer. While much evidence has suggested that OPN is associated with cancer, its functional contribution to cancer remains poorly understood. Here we will review evidence for mechanisms by which OPN may act to enhance malignancy, including evidence that signaling pathways directly induced by OPN, as well as interactions with growth factor receptor pathways, can combine to activate expression of genes and functions that contribute to metastasis. OPN has been shown to be over-expressed in a variety of human tumors and is present in elevated levels in the blood of some patients with metastatic cancers. We also will discuss recent clinical evidence that suggests that OPN is not only associated with several tumor types, but that levels of OPN in cancer patients' blood or tumors may provide prognostic information. PMID- 11899238 TI - Vehicles for genetic vaccines against human immunodeficiency virus: induction of T cell-mediated immune responses. AB - Success of a candidate vaccine against human immunodeficiency virus (HIV) depends on the type, site, strength, longevity and specificity of the immune responses it induces. The specificity of a vaccine is determined by the HIV-derived immunogens it employs in its formulation. Central to the other features is a correct and efficient delivery of the immunogens to the relevant cells of the immune system, which leads to orchestrated actions of millions of cells of several types and functions at multiple sites in the body. Thus, for elicitation of cytotoxic T lymphocyte responses, immunogens have to be delivered to the so called 'professional' antigen-presenting cells in a way that leads to a specific activation and expansion of naive or precursor T cells, subsequent maturation of effector functions and, importantly, generation of a potent immunological memory. Many aspects of theseprocesses are currently unknown. However, it is very likely that the immunogenicities of genetic vaccines, i.e. vaccines delivering genes coding for immunogens rather than purified possibly adjuvanted proteins or peptides themselves, are in great part determined by the choice of vaccine vehicles and route of administration. In addition, vaccine immunogenicities can be augmented semi-rationally by immunogen engineering and co-delivering immunomodulatory molecules, and empirically by combining different vehicles expressing the same immunogen in heterologous prime-boost protocols. In any case, a successful vaccination strategy against HIV as well as other chronic viral infections has to elicit better immune responses than the natural infections do. PMID- 11899237 TI - Lessons from HIV: movement of macromolecules inside the cell. AB - Molecular biological investigations of HIV have made fundamental contributions to our understanding of eukaryotic biology. These studies elucidated new paradigms in transcription, RNA and protein export from the nucleus to the cytoplasm, cellular activation, morphology and vesicular trafficking. PMID- 11899240 TI - Microdissection and the study of cancer pathways. AB - Abstract: The study of genetic alterations in tumors and their precursor lesions is often hampered by the presence of a heterogeneous background of non-neoplastic elements such as stromal cells, inflammatory cells, and angiogenic elements. Microdissection involves the extraction of specific populations of cells under direct visualization. In this article, we will discuss the currently available techniques of microdissection, and briefly review how this material is being utilized in the study of cancer pathways. Microdissected tissue is amenable for the study of cancer genomics, expression analysis and most recently, cancer proteomics. The purity of reagents obtained from microdissected material has resulted in the successful identification of tumor suppressor genes as well as novel transcripts and proteins that are altered in neoplastic cells. Improved techniques of tissue fixation and microdissection, supplemented with ancillary technology such as pre-amplification, have permitted the use of increasingly smaller quantities of material for the study of cancer pathways. Importantly, it is now possible to analyze many of the genetic changes that precede cancer, thereby identifying populations "at risk" for developing malignancies in the future. PMID- 11899239 TI - Natural T cell immunity to intracellular pathogens and nonpeptidic immunoregulatory drugs. AB - Natural T (NT) lymphocytes recognize infected cells or microbial compounds without the classical genetic restriction of polymorphic major histocompatibility complex (MHC) molecules. This innate recognition pathway results in a broad and rapid antimicrobial response that may be critical for controlling the spread of intracellular pathogens, requiring the elimination of the infecting agent from both extracellular spaces and host cells. NT cells are mainly composed of alphabeta and gammadelta T lymphocytes that express natural killer (NK) receptors and recognize preferentially various nonpeptidic antigens. Similar to NK cells, NT lymphocytes can 'see' and kill target cells deficient in the expression of one or more MHC class I molecules. NT cells expressing the alphabeta TCR can recognize lipid and lipoglycan antigens presented in the context of nonpolymorphic CD1 molecules, whereas phosphocarbohydrates and akilamines induce constitutive responses in most Vgamma9Vdelta2 NT lymphocytes. The remaining fraction of gammadelta NT cells express the Vdelta1 chain associated with different Vgamma-chains and may directly recognize self-antigens such as MICA, MICB or CD1 molecules. It is possible that NT lymphocytes may play two opposite roles during intracellular infections. First, in the acute phase, they may be critical for the initiation of pathogen elimination. Second, in the chronic phase, NT cells may be dangerous, if their potential autoreactivity is not well controlled. It is conceivable that novel strategies of immune intervention against emerging and re-emerging intracellular pathogens, such as human immundeficiency virus (HIV), hepatitis-C virus (HCV) and Mycobacterium tuberculosis (MTB) may involve the control of NT cell activation/anergy by (nonpeptidic) immunoregulatory drugs. PMID- 11899242 TI - The human ATP-binding cassette transporter genes: from the bench to the bedside. AB - ATP-binding cassette (ABC) transporter genes are ubiquitously present in most organisms from bacteria to man. This gene family is the largest one known as of yet. Still growing, the number of human ABC transporters counts currently 47 members which belong to seven subfamilies. ABC transporters share a similar molecular architecture: (1) Full-structured transporters harbor two symmetric halves each consisting of one nucleotide binding domain (NBD) and one transmembrane domain (TMD). (2) Half-transporters with one NBD and one TMD homo- or heterodimerize to functional transporter complexes. ABC transporters are "traffic ATPases" which hydrolyze ATP and which transport a wide array of molecules or conduct the transport of molecules by stimulating other translocation mechanisms. Many ABC transporters are involved in human inherited or sporadic diseases such as cystic fibrosis, adrenoleukodystrophy, Stargardt's disease, drug-resistant tumors, Dubin-Johnson syndrome, Byler's disease, progressive familiar intrahepatic cholestasis, X-linked sideroblastic anemia and ataxia, persistent hyperinsulimenic hypoglycemia of infancy, and others. The present review summarizes the current findings in basic research and the efforts for bridging the gap to clinical applications in therapy and diagnostics. PMID- 11899241 TI - The molecular basis of type 1 glycogen storage diseases. AB - Glycogen storage disease type 1 (GSD-1), also known as von Gierke disease, is a group of autosomal recessive metabolic disorders caused by deficiencies in the activity of the glucose-6-phosphatase (G6Pase) system that consists of at least two membrane proteins, glucose-6-phosphate transporter (G6PT) and G6Pase. G6PT translocates glucose-6-phosphate (G6P) from cytoplasm to the lumen of the endoplasmic reticulum (ER) and G6Pase catalyzes the hydrolysis of G6P to produce glucose and phosphate. Therefore, G6PT and G6Pase work in concert to maintain glucose homeostasis. Deficiencies in G6Pase and G6PT cause GSD-1a and GSD-1b, respectively. Both manifest functional G6Pase deficiency characterized by growth retardation, hypoglycemia, hepatomegaly, kidney enlargement, hyperlipidemia, hyperuricemia, and lactic acidemia. GSD-1b patients also suffer from chronic neutropenia and functional deficiencies of neutrophils and monocytes, resulting in recurrent bacterial infections as well as ulceration of the oral and intestinal mucosa. The G6Pase gene maps to chromosome 17q21 and encodes a 36-kDa glycoprotein that is anchored to the ER by 9 transmembrane helices with its active site facing the lumen. Animal models of GSD-1a have been developed and are being exploited to delineate the disease more precisely and to develop new therapies. The G6PT gene maps to chromosome 11q23 and encodes a 37-kDa protein that is anchored to the ER by 10 transmembrane helices. A functional assay for the recombinant G6PT protein has been established, which showed that G6PT functions as a G6P transporter in the absence of G6Pase. However, microsomal G6P uptake activity was markedly enhanced in the simultaneous presence of G6PT and G6Pase. The cloning of the G6PT gene now permits animal models of GSD-1b to be generated. These recent developments are increasing our understanding of the GSD l disorders and the G6Pase system, knowledge that will facilitate the development of novel therapeutic approaches for these disorders. PMID- 11899243 TI - Gene therapy for lung diseases: development in the vector biology and novel concepts for gene therapy applications. AB - The lung represents an attractive target organ for somatic gene therapy strategy in that, (1) it is easily accessible by vectors, (2) most frequent hereditary disorders, cystic fibrosis (CF) and alpha1-antitrypsin deficiency (alpha1AT), occur in the lung, and (3) carcinoma of the lung is apparently a most common cause of death in humans. To date, approximately 400 clinical protocols for human gene therapy have been approved, and approximately 10% of the protocols target lung diseases such as cystic fibrosis (CF) and lung cancer. Currently available data from some of these human trials have successfully demonstrated that gene transfer to the human lung is possible, and that the strategy of overexpressing exogenous genes for curing or controlling lung diseases is potentially promising. In this manuscript, focusing on gene therapy of lung disorders, we aim to give an overview of the hurdles of current gene transfer strategies to overcome, then and also we aim to review recent, remarkable progresses in the vector biology that are potentially promising to maximize safety and efficiency of gene therapy. In addition, based on the most recent advances in the understanding of the molecular biological aspects of the pathogenesis of lung cancer, asthma, pulmonary fibrosis, and acute lung injury, novel therapeutic strategies of gene therapy for inflammatory and malignant diseases of the lung are discussed. PMID- 11899244 TI - Molecular genetics of left ventricular dysfunction. AB - The left ventricle (LV) plays a central role in the maintenance of health of children and adults due to its role as the major pump of the heart. In cases of LV dysfunction, a significant percentage of affected individuals develop signs and symptoms of congestive heart failure (CHF), leading to the need for therapeutic intervention. Therapy for these patients include anticongestive medications and, in some, placement of devices such as aortic balloon pump or left ventricular assist device (LVAD), or cardiac transplantation. In the majority of patients the etiology is unknown, leading to the term idiopathic dilated cardiomyopathy (IDC). During the past decade, the basis of LV dysfunction has begun to unravel. In approximately 30-40% of cases, the disorder is inherited; autosomal dominant inheritance is most common (although X-linked, autosomal recessive and mitochondrial inheritance occurs). In the remaining patients, the disorder is presumed to be acquired, with inflammatory heart disease playing an important role. In the case of familial dilated cardiomyopathy (FDCM), the genetic basis is beginning to unfold. To date, two genes for X-linked FDCM (dystrophin, G4.5) have been identified and four genes for the autosomal dominant form (actin, desmin, lamin A/C, delta-sarcoglycan) have been described. In one form of inflammatory heart disease, coxsackievirus myocarditis, inflammatory mediators and dystrophin cleavage play a role in the development of LV dysfunction. In this review, we will describe the molecular genetics of LV dysfunction and provide evidence for a "final common pathway" responsible for the phenotype. PMID- 11899245 TI - Genetics of type 2 diabetes: insight from targeted mouse mutants. AB - Diabetes affects millions of people worldwide, and its chronic complications are a leading cause of death in many industrialized countries. In a minority of patients, diabetes is brought about by the auto-immune destruction of insulin producing pancreatic beta cells (Type 1 diabetes). In the vast majority of patients, diabetes is brought about by a combination of genetic and environmental factors that affect the organism's ability to respond to insulin (Type 2 diabetes). This impairment is due to a complex abnormality involving insulin action at the periphery and insulin production in the beta cell. Genetic factors play a key role in the development of type 2 diabetes. However, the inheritance of diabetes is non-Mendelian in nature, due to genetic heterogeneity, polygenic pathogenesis and incomplete penetrance. For these reasons, many laboratories have developed "designer" mice bearing targeted mutations in genes of the insulin action and insulin secretion pathways in order to develop a better model for the inheritance and pathogenesis of type 2 diabetes. These mutant mice are beginning to challenge established paradigms in the pathogenesis of type 2 diabetes and to shed light onto the genetic interactions underlying its complex inheritance. Here we review recent progress in the field and assess its impact on human studies of the genetics, prevention and treatment of type 2 diabetes. PMID- 11899246 TI - Mitochondria in apoptosis and human disease. AB - Apoptosis is a process of cell suicide whereby individual cells are destroyed while preserving the integrity and architecture of surrounding tissue. This targeted cell destruction is critical both in physiological contexts as well as pathological states. It seems increasingly evident that mitochondria play an important role in the regulation of programmed cell death via release of proapoptotic agents and/or disruption of cellular energy metabolism. The mechanisms of mitochondrial involvement are beginning to be elucidated, and may involve the participation of bcl-2 family members, reactive oxygen species, and caspases. As part of a central mechanism of amplification of the apoptotic signal, mitochondria may be an appropriate target for therapeutic agents designed to modulate apoptosis. This review focuses on recent advances in understanding mitochondrial mechanisms in apoptosis and the involvement of these pathways in human disease. PMID- 11899248 TI - Discovery of the factor Xa inhibitor, ZK 807834 (CI-1031). AB - Discoveries that lead to ZK 807834 (CI-1031, 2a), a potent and selective factor Xa (fXa) inhibitor currently in clinical testing as an intravenous antithrombotic, were initiated by the identification of the potent (Z,Z)-isomer of BABCH (1c). A structure-activity relationship (SAR) was established with a series of analogues of BABCH. This SAR database, combined with computer modeling, demonstrated that binding of the second basic group in the S3/S4 pocket provided fXa potency and that a carboxylic acid group on the opposite side of the molecule resulted in selectivity versus thrombin. Simple substitution of a cyclic urea for the unsaturated ketone structure of BABCH gave disappointing results, but discovery of the bisphenoxy-pyridine analogues provided a template that could be readily optimized. The SAR established for this template is described and compared with computer modeling, REDOR NMR and X-ray crystallography studies. Inhibitor binding to fXa was increased by the introduction of a hydroxyl group on the proximal phenylamidine ring and by the introduction of fluorine atoms at C-3 and C-5 of the pyridine ring. Pharmacokinetic parameters were improved by balancing the contributions from the substituents on the distal ring and the central pyridine ring. The optimal combination was a methyl-(2H)-imidazoline group on the distal ring and a sarcosine at C-4 of the pyridine ring. The promising preclinical database for CI-1031 is described. This review relates the SAR leading to the discovery of the clinical candidate, CI-1031 directly to our best understanding of how this potent inhibitor interacts with the fXa active site. PMID- 11899247 TI - Discovery of transition state factor Xa inhibitors as potential anticoagulant agents. AB - Factor Xa is an attractive biological target in the discovery and development of either parenteral or orally active anticoagulant agents. Several strategies have been utilized at COR Therapeutics in the pursuit of tri-peptide based transition state mimetic factor Xa inhibitors with high aqueous solubility. Some of these inhibitors have displayed excellent in vitro potency in inhibiting factor Xa in the prothrombinase complex. More importantly, these compounds showed strong in vivo antithrombotic efficacy without significant bleeding complications in several animal thrombosis models. These results demonstrated that small molecule factor Xa inhibitors could be advantageous over Warfarin and LMWH. For the discovery and development of orally active anticoagulant agents, small organic molecules as reversible factor Xa inhibitors were explored. From a medicinal chemistry perspective, significant insight has been gained regarding the in vivo antithrombotic efficacy and pharmacokinetic behaviors of each class of factor Xa inhibitors. This review will focus on the design and discovery of transition state factor Xa inhibitors as potential parenteral anticoagulant agents. Several excellent comprehensive review articles on factor Xa inhibitors have appeared recently [1-4]. PMID- 11899251 TI - The use of 3D structural data in the design of specific factor Xa inhibitors. AB - Factor Xa (fXa) is a serine protease that plays a critical role in the blood coagulation process and qualifies as an attractive target for finding new antithrombotics. In the case of fXa several structure based drug design strategies have been followed because of the difficulty in growing fXa co crystals routinely. This has led to the use of surrogate proteins such as trypsin. Factor Xa inhibitors for which the binding mode has been determined experimentally or modeled are described in this review. The inhibitors are divided into three fragments: a P1 group, a central scaffold and a P4 group. In this review, interactions in each sub-site of fXa with various inhibitor fragments have been examined at the molecular level and were shown to bind, in most cases, independently of the rest of the molecule. Knowledge of the 3D structure of the binding mode of ligands to target proteins has been successfully applied in designing fXa inhibitors with enhanced specificity, affinity and has provided hints to modulate the physico-chemical properties of the small molecule ligand. PMID- 11899249 TI - The design and synthesis of noncovalent factor Xa inhibitors. AB - Thrombosis is a major cause of mortality in the industrialized world. Therefore, the control of blood coagulation has become a major target for new therapeutic agents. One attractive approach is the inhibition of factor Xa (fXa), the enzyme directly responsible for thrombin generation. In this review we describe our approaches in the design and synthesis of small molecule, noncovalent fXa inhibitors. Rational drug design and selective screening of our GPIIb/IIIa library afforded several lead compounds for our fXa program. Following-up the leads in the isoxazoline series led to potent fXa inhibitors such as SF303 and SK509 with only one basic group. The isoxazole series was then designed to remove the chiral center in the isoxazoline ring, and this effort led to SA862 which has subnanomolar fXa affinity. Optimizing the core structure generated a series of novel five-membered ring heterocycles substituted with benzamidine, which are potent fXa inhibitors. Further optimization in the pyrazole series resulted in the discovery of fXa inhibitors such as SN429 with picomolar fXa affinity. Efforts to improve the oral bioavailability by lowering the basicity of these compounds, while simultaneously maintaining potency against fXa, culminated in the discovery of DPC 423. DPC 423 was selected for clinical evaluation as a potent and orally bioavailable fXa inhibitor. PMID- 11899250 TI - Coagulation factor Xa inhibition: biological background and rationale. AB - Ischemic heart disease and cerebrovascular disease are the leading causes of death in the world. Surprisingly, these diseases are treated by relatively antiquated drugs. However, due to our improved understanding of the underlying pathology of these diseases, and a number of technological advances in tools for drug discovery and chemical optimization, an exciting new wave of antithrombotic compounds is beginning to emerge in clinical trials. These agents, referred to as direct coagulation factor Xa inhibitors, appear to provide an enhanced risk benefit margin compared to conventional therapy. Preclinical and early clinical data gathered over the past few years suggests that direct fXa inhibitors will provide the necessary advancements in efficacy, safety, and ease of use required to displace conventional therapy. Whether or not these agents will succeed will be determined as this class of agents advances through clinical trials in the near future. This review describes some of the key studies that sparked an interest in fXa as a therapeutic target, highlighting the findings that provided important rationale for continuing the development of potent and selective direct fXa inhibitors. PMID- 11899252 TI - The design of competitive, small-molecule inhibitors of coagulation factor Xa. AB - The last five years have seen an explosion of research into inhibitors of Factor Xa as potential antithrombotic agents. Aventis Pharma was a participant in this effort and its two founder companies have substantially contributed to the discovery of new inhibitors over the years. This review traces the systematic development of the former Rhone-Poulenc Rorer factor Xa program from conception to the realization of potent, orally bioavailable inhibitors with exquisite selectivity against other serine proteases. The work on beta-aminoesters described in Part 1 culminates in the development of FXV673 (Ki = 0.5 nM), an effective anticoagulant for acute indications. Part 2.2 details the de novo design of the pyrrolidinone series of inhibitors (RPR120844), within which a group of efficacious i.v. agents were identified (e.g. RPR130737, Ki = 2 nM). The first active and bioavailable benzamidine isostere i.e. the 1-aminoisoquinoline (RPR208815, Ki = 22 nM) was discovered on the pyrrolidinone scaffold (Part 2.3). Ultimately a variety of benzamidine mimics were explored and incorporated into the ketopiperazine series; the 6-substituted aminoquinazolines were found to be the most potent (Part 3). The azaindole, as represented by RPR200443 (Ki = 4 nM), stands out as imparting favorable pharmacokinetic properties to the sulfonamido ketopiperazines. PMID- 11899253 TI - Lipoprotein cholesterol and atherosclerosis. AB - Progressive accumulation of cholesterol in the arterial wall causes atherosclerosis, the pathologic process underlying most heart attacks and strokes. Low density lipoprotein (LDL), the major carrier of blood cholesterol, has been implicated in the buildup of cholesterol in atherosclerotic plaques. Endothelial cells that line arteries function to transport LDL into the vessel wall. Models for the mechanism of cholesterol accumulation in atherosclerotic plaques emphasize increased LDL uptake into the vessel wall or increased retention of LDL that has entered the vessel wall. This article reviews the pathways of cholesterol entry and removal, the metabolism, and the physical changes of cholesterol in the vessel wall. How these processes are believed to contribute to cholesterol buildup in atherosclerotic plaques is discussed. PMID- 11899254 TI - In search of pathogenic mechanisms in endometriosis: the challenge for molecular cell biology. AB - Endometriosis, defined histologically as the presence of endometrium-like glands and stroma outside the uterus, is a chronic, invasive and metastasising disease. It shares features with malignant tumours (invasion and metastasis) but is not neoplastic. Despite the fact that endometriosis is one of the most frequent gynaecological diseases, it is under researched, puzzling and highly debated. The aetiology and pathogenesis is little understood although it is agreed that implantation, at least in many cases, is responsible for endometriosis. This theory advocates retrograde menstruation as the underlying phenomenon, where cells of the menstrual efflux provide the cellular source for endometriotic lesion formation. Causative therapy and non-invasive diagnostics of endometriosis do not exist. Thus, there is a substantial but unmet need for molecular and cellular research to unravel the pathogenic mechanisms of endometriosis as a basis for developing novel diagnostic and therapeutic concepts. In this review, we specifically focus on the cellular basis of lesion formation, the possible modulation of this by cytokines and other factors and the characteristics of endometriotic cells in terms of invasion and metastasis. Considering available experimental information, we concentrate on arguments and ideas in favour of an endometriotic founder cell population exhibiting substantial plasticity for differentiation and self-renewal. Perhaps present in the menstrual efflux or arising by metaplasia (a complementary theory to implantation), this cell type might respond to stimuli present in the ectopic host environment and establish the endometriotic phenotype. PMID- 11899255 TI - The role of the ubiquitin-proteasome pathway in MHC class I antigen processing: implications for vaccine design. AB - Proteasomes are multisubunit enzyme complexes that reside in the cytoplasm and nucleus of eukaryotic cells. By selective protein degradation, proteasomes regulate many cellular processes including MHC class I antigen processing. Three constitutively expressed catalytic subunits are responsible for proteasome mediated proteolysis. These subunits are exchanged for three homologous subunits, the immunosubunits, in IFNgamma-exposed cells and in cells with specialized antigen presenting function. Both constitutive and immunoproteasomes degrade endogenous proteins into small peptide fragments that can bind to MHC class I molecules for presentation on the cell surface to cytotoxic T lymphocytes. However, immunoproteasomes seem to fulfill this function more efficiently. IFNgamma further induces the expression of a proteasome activator, PA28, which can also enhance antigenic peptide production by proteasomes. In this review, we will introduce the ubiquitin-proteasome system and summarize recent findings regarding the role of the IFNgamma-inducible proteasome subunits and proteasome regulators in antigen processing. We review the different ways by which tumors and viruses have been found to target the proteasome system to avoid MHC class I presentation of their antigens, and discuss recent progressions in the development of computer assisted approaches to predict CTL epitopes within larger protein sequences, based on proteasome cleavage specificity. The availability of such programs as well as a general insight into the proteasome mediated steps in MHC class I antigen processing provides us with a rational basis for the design of new antiviral and anticancer T cell vaccines. PMID- 11899256 TI - Molecular mechanisms of neuronal migration disorders, quo vadis? AB - Following terminal mitosis, neuronal precursor cells leave their site of origin and migrate towards their definitive site of residency. In order to establish the intricate cytoarchitecture described in the adult human brain, neuronal migration must be finely regulated. In humans, brain malformations can result from neuronal migration defects. The spectrum of migration disorder severity extends from few heterotopic neurons, as observed in periventricular heterotopia, to a complete cortical disorganization, as observed in cases of lissencephaly. Recently, specific migration disorders have been linked to mutations/deletions in the doublecortin, filamin-1, LIS1 and reelin genes. These proteins act at different levels of the signaling cascades transducing extracellular guiding cues into cytoskeletal reorganization. Here, we summarize the data concerning these four molecules and speculate on their functions and interaction partners during neuronal development. PMID- 11899257 TI - The molecular basis of lymphoid architecture and B cell responses: implications for immunodeficiency and immunopathology. AB - Immune responses usually take place in secondary lymphoid organs such as spleen and lymph nodes. Most lymphocytes within these organs are in transit, yet lymphoid organ structure is highly organized; T and B cells segregate into separate regions. B cell compartments include naive cells within follicles, marginal zones and B-1 cells. Interactions between TNF family molecules on hematopoietic cells and their receptors on mesenchymal cells guide the initial phase of lymphoid organogenesis, and regulate chemokine secretion that mediates subsequent T-B cell segregation. Recruitment of B cells into different compartments depends on both the milieu established during organogenesis, and the threshold for B cell receptor signaling, which is modulated by numerous coreceptors. Novel intrafollicular (germinal center) and extrafollicular (plasma cell) compartments are established when B cells respond to antigen. These divergent B cell responses are mediated by different patterns of gene expression, and influenced again by BCR signaling threshold and cellular interactions that depend on normal lymphoid architecture. Aberrant B cell responses are reviewed in the light of these principles taking into account the molecular and architectural aspects of immunopathology. Histological features of immunodeficiency reflect defects of B cell recruitment or differentiation. B cell hyper-reactivity may arise from altered BCR signaling thresholds (autoimmunity), defects in stimuli that guide differentiation in response to antigen (follicular hyperplasia vs plasmacytosis), or defective B cell gene expression. Interestingly, in diseases such as rheumatoid arthritis, Sjogren's syndrome and Hashimoto's thyroiditis lymphoid organogenesis may be recapitulated in non-lymphoid parenchyma, under the influence of molecular interactions similar to those that operate during embryogenesis. PMID- 11899258 TI - The role of pancreatic chromogranins in islet physiology. AB - Chromogranins are acidic secretory glycoproteins with a widespread but specific distribution in neuroendocrine tissues. The chromogranin family is heterogenous, consisting of propeptides such as chromogranin-A, chromogranin-B and secretogranin II, which can either elicit an effect themselves, or serve as precursors to a large number of peptides, which are biologically more active. Chromogranin processing varies in different neuroendocrine tissues. Furthermore, it is more marked in pancreatic islets than in many other tissues. Chromogranin-A and chromogranin-B are expressed in all types of pancreatic islet cells, whereas secretogranin II has not been found in pancreatic tissue. The aim of the present mini review is to focus on chromogranin-A, chromogranin-B and their derived peptides, in the function of pancreatic islets. PMID- 11899259 TI - Beta-amyloid, neuronal death and Alzheimer's disease. AB - Alzheimer's disease (AD) is a common neurodegenerative disease that affects cognitive function in the elderly. Large extracellular beta-amyloid (Abeta) plaques and tau-containing intraneuronal neurofibrillary tangles characterize AD from a histopathologic perspective. However, the severity of dementia in AD is more closely related to the degree of the associated neuronal and synaptic loss. It is not known how neurons die and synapses are lost in AD; the current review summarizes what is known about this issue. Most evidence indicates that amyloid precursor protein (APP) processing is central to the AD process. The Abeta in plaques is a metabolite of the APP that forms when an alternative (beta-secretase and then gamma-secretase) enzymatic pathway is utilized for processing. Mutations of the APP gene lead to AD by influencing APP metabolism. One leading theory is that the Abeta in plaques leads to AD because Abeta is directly toxic to the adjacent neurons. Other theories advance the notion that neuronal death is triggered by intracellular events that occur during APP processing or by extraneuronal preplaque Abeta oligomers. Some investigators speculate that in many cases there is a more general disorder of protein processing in neurons that leads to cell death. In the later models, Abeta plaques are a byproduct of the disease process, rather than the direct cause of neuronal death. A direct correlation between Abeta plaque burden and neuronal (or synaptic) loss should occur in AD if Abeta plaques cause AD through a direct toxic effect. However, histopathologic studies indicate that the correlation between Abeta plaque burden and neuronal (or synaptic) loss is poor. We conclude that APP processing and Abeta formation is important to the AD process, but that neuronal alterations that underlie symptoms of AD are not due exclusively to a direct toxic effect of the Abeta deposits that occur in plaques. A more general problem with protein processing, damage due to the neuron from accumulation of intraneuronal Abeta or extracellular, preplaque Abeta may also be important as underlying factors in the dementia of AD. PMID- 11899260 TI - Mammalian secreted phospholipases A2 and their pathophysiological significance in inflammatory diseases. AB - Phospholipases A2 (PLA2s) represent a growing family of enzymes that catalyze the hydrolysis of phospholipids at the sn-2 position leading to the generation of free fatty acids and lysophospholipids. Mammalian PLA2s are divided into two major classes according to their molecular mass and location: intracellular PLA2 and secreted PLA2 (sPLA2). Type-IIA sPLA2 (sPLA2-IIA), the best studied enzyme of sPLA2, plays a role in the pathogenesis of various inflammatory diseases. Conversely, sPLA2-IIA can exert beneficial action in the context of infectious diseases since recent studies have shown that this enzyme exhibits potent bactericidal effects. Induction of the synthesis of sPLA2-IIA is generally initiated by endotoxin and a limited number of cytokines via paracrine and/or autocrine processes. If the mechanisms involved in the regulation of sPLA2-IIA gene expression have been relatively clarified, little is known on the mechanisms that regulate the expression of other sPLA2. There have been substantial progresses in understanding the transcriptional regulation of sPLA2-IIA expression. Recently, transcription factors including NF-kappaB, PPAR, C/EBP have been identified to play a prominent role in the regulation of sPLA2-IIA gene expression. The activation of these transcription factors is under the control of distinct signaling pathways (PKC, cAMP ...). Accumulating evidences in the literature suggest that cytosolic PLA2 together with two sPLA2 isozymes (sPLA2 IIA and sPLA2-V) are functionally coupled with cyclooxygenase-1 and 2 pathways, respectively, for immediate and delayed PG biosynthesis. This spatio-temporal coupling of cyclooxygenase enzymes with PLA2s may represent a key mechanism in the propagation of inflammatory reaction. Unraveling the mechanisms involved in the regulation of the expression of sPLA2s is important for understanding their pathophysiological roles in inflammatory diseases. PMID- 11899261 TI - Progress in gene delivery by cationic lipids: guanidinium-cholesterol-based systems as an example. AB - Artificial self-assembling systems are currently widely investigated as an alternative approach to recombinant viruses for gene transfection in vitro and in vivo. Cationic lipids are particularly attractive, as a great variety of well characterized reagents can be synthesized from there. Over the last few years, numerous cationic lipid systems have been developed and shown to be efficient for in vitro transfection. However, although some promising results have been reported in the in vivo setting (even in clinical gene therapy trials in man), the in vivo use of cationic lipid-based systems is still problematic, especially when considering the systemic route of administration. Herein, we summarize our own research on a particular class of cationic lipids, cholesterol derivatives characterized by polar headgroups with guanidinium functions, in order to illustrate the basic principles of and the positive results already obtained by cationic lipid-mediated gene delivery as well as the remaining problems that need to be urgently resolved, particularly as regards the systemic administration. In this forward-looking review, we also discuss the present efforts to develop modular systems for improved in vivo transfection. Indeed, lipid-based vectors offer the possibility to create sophisticated modular gene delivery systems capable of self-assembly via hydrophobic interaction between their components, the role of the different functional elements being to help in overcoming the distinct extracellular and cellular barriers to in vivo gene transfection into the various somatic target tissues. PMID- 11899262 TI - Sodium lauryl sulfate, a microbicide effective against enveloped and nonenveloped viruses. AB - The number of individuals infected with human immunodeficiency virus (HIV) and other pathogens causing sexually transmitted diseases (STDs) is growing dramatically worldwide. Globally, heterosexual transmission may account for as much as 85-90% of new cases of HIV infection. Latex condoms represent an effective barrier against sexually transmitted pathogens, but unfortunately, their use is not generalized. Therefore, there is an urgent need to develop safe and potent topical microbicides under the control of women to efficiently reduce the spread of sexually transmitted infections. Sodium lauryl sulfate (SLS), an anionic surfactant with protein denaturing potency, is a potent inhibitor of the infectivity of several enveloped (Herpes simplex viruses, HIV-1, Semliki Forest virus) and nonenveloped (papillomaviruses, reovirus, rotavirus and poliovirus) viruses. The mechanism of action of SLS involves the solubilization of the viral envelope and/or the denaturation of envelope and/or capsid proteins. Studies have shown that SLS is not toxic for cultured cell lines of different origins at concentrations that inactivate HIV-1, herpes and human papillomavirus in vitro. In addition, intravaginal pretreatment of mice with a gel formulation containing SLS, completely protected animals against Herpes simplex virus type-2 infection. The gel formulation containing SLS was also well-tolerated following repeated intravaginal administrations to rabbits. Taken together, these data suggest that SLS represents a potential candidate for the use as a topical microbicide to prevent the sexual transmission of HIV-1, herpes, human papillomavirus and possibly other sexually transmitted pathogens. The impact of such a preventive tool on public health can be enormous. PMID- 11899263 TI - Studies on target genes of general anesthetics. AB - Generally speaking, we cannot fully understand the mechanisms of general anesthesia until the molecular mechanisms of consciousness are fully elucidated. Loss of consciousness induced by general anesthetics might involve sensation, motor activity, behavior, memory and self-consciousness. The effects of many anesthetics are not limited to humans but also extend to the animals. Similar levels of minimum anesthetic concentrations are required to induce anesthesia in animals and human, i.e., the minimum alveolar concentration (MAC). Such similarity probably reflects identical anesthetic target molecules and functional conservation based on gene conservation. Thus, to study the mechanisms of anesthetic action, various animal models that are accessible to genetic manipulation, such as nematodes (Caenorhabditis elegans), fruit flies (Drosophila) and mice can be used. Genetic techniques allow for the rapid identification and characterization of genes involved in the actions of general anesthetics. In this review, I will describe the genetic mutations and putative target genes of general anesthetics. PMID- 11899264 TI - Chronic tendon pain: no signs of chemical inflammation but high concentrations of the neurotransmitter glutamate. Implications for treatment? AB - Chronic musculo-tendinous pain syndromes are relatively common and associated with very high socio-economic costs. Their aetiology and pathogenesis are still unknown. In the athletic population, chronic tendon pain is most often seen among recreational male and female athletes in the age group between 30-60 years, and is considered to be associated with overuse of the aged tendon. Treatment is known to be difficult. In general, these chronic painful conditions have been considered to include an inflammatory component, and the nomenclature used (tendinitis, tendonitis) most often implies an inflammatory involvement. Despite that tendon biopsies have shown an absence of inflammatory cell infiltration, anti-inflammatory agents (NSAID'S, corticosteroidal injections) are most often included in the treatment. Our research has been focused on chronic painful conditions in the Achilles-, patellar-, and extensor carpi radialis brevis (ECRB) tendons. We have demonstrated, for the first time, that it is possible to use the microdialysis technique for in vivo investigations of human tendons, and that the excitatory neurotransmitter glutamate exists in human tendons. We have identified and measured the concentrations of prostaglandin E2 (PGE2) in tendons as well. The results showed that chronic painful tendinosis (Achilles-, patellar-, ECRB-) had significantly higher concentrations of glutamate, but not PGE2, as compared to the pain-free normal tendons. With the use of immunohistochemical analyses and enzyme histochemistry of human Achilles- and patellar tendon biopsies, we have also, for the first time demonstrated that glutamate NMDARI-immunoreaction was present in nerve structures. These findings altogether, indicate that glutamate might be involved in chronic tendon pain, and further emphasizes that there is no chemical inflammation (normal PGE2 levels) in the chronic stage of these relatively common so-called tendinopathies. The findings of glutamate and it's NMDARI-receptors might have implications for treatment and be a potential target for drugs. PMID- 11899267 TI - Photonic encoder and decoder for optical code-division multiplexing with time-to space converters and angle-multiplexed holograms. AB - A photonic encoder-decoder pair for optical code-division multiplexing (OCDM) that uses time-to-space converters and angle-multiplexed holograms is proposed. The encoder converts the pulse from each input port into a specific code and multiplexes input signals into the output port. The hologram in the decoder generates a correlation waveform between the transmitted code and the recorded code. The performance of the OCDM system with the encoder-decoder pair is estimated. The maximum spectral efficiency for 8-bit length orthogonal codes in the worst case at a bit-error rate of 10(-9) is 0.17 (bits/s)/Hz when the number of channels is 8. PMID- 11899265 TI - Hypericin--a new antiviral and antitumor photosensitizer: mechanism of action and interaction with biological macromolecules. AB - Hypericin, a naturally occurring pigment, is found in certain species of plants from the genus Hypericum, the most common of which is Saint John's Wort (Hypericum perforatum). Recent interest in hypericin is provoked by the discovery that it possesses extremely high toxicity towards certain viruses notably the class of enveloped viruses that includes human immunodeficiency virus (HIV) and toward tumors, and that this toxicity absolutely requires light. Consequently, a detailed understanding of the interaction of hypericin with cellular components (membranes, proteins, nucleic acids) and with light is of fundamental biological importance. The antiviral and antineoplastic activities of hypericin and its derivatives and its mode of action have been widely studied, in the last two decades. This review is focused on the results obtained in the study of hypericin heteroassociations with biological macromolecules, DNA and human serum albumin in particular. An alternative type of the hypericin photosensitizing activity associated with its ability to produce a photogenerated pH drop is discussed that and discussed in connection with its potential application in photodynamic therapy. In the review, it is also presented how an interdisciplinary approach supported by sophisticated techniques of optical spectroscopy and molecular modeling can be effectively applied for the identification of the specific binding sites of the drug in some biomacromolecules as well as for the determination of the physico-chemical mechanism'of the biological activity of hypericin. PMID- 11899266 TI - Fan-out diffractive optical elements designed for increased fabrication tolerances to linear relief depth errors. AB - The intensity uniformity of the spots generated by fan-out diffractive optical elements (DOEs) (or kinoforms) is often highly sensitive to any fabrication error that leads to a deviation of the surface-relief depth of the DOE from its design value. Many of the fabrication errors, such as those that are due to insufficient control of development or etch rates, increase almost linearly with the desired relief depth in every position of the DOE. We present an algorithm for designing fan-out DOEs with a significantly reduced sensitivity of the intensity uniformity to such errors. The reduced sensitivity can be obtained without reducing the efficiency of the DOE. Experimental results for fabricated DOEs show that reduced sensitivity is also obtained in practice. PMID- 11899268 TI - Multiplexed computer-generated hologram with polygonal apertures. AB - A novel type of multiplexed computer-generated hologram (CGH) is designed with more than one billion of pixels per period. It consists of elementary cells divided into arbitrary-shaped polygonal apertures, the division being identical in all cells. The cells are further digitized into pixel arrays to exploit the huge space-bandwidth product of electron-beam lithography. The polygonal apertures in the same location inside the cells constitute a subhologram. With the Abbe transform that has never, to our knowledge, been used in other CGH designs, the subhologram images (subimages) are obtained with fast Fourier transforms. It is therefore possible to design a multiplexed CGH that has a size thousands of times larger than the manageable size of a conventional CGH designed with the iterative Fourier transform algorithm (IFTA). A much larger object window than that of the conventional CGH can also be achieved with the multiplexed polygonal-aperture CGH, owing to its extremely large dimensions. The multiplexed polygonal-aperture CGH is designed with the novel iterative subhologram design algorithm, which considers the coherent summation of the subimages and applies constraints on the total image, subimages, and subholograms. As a result, the noise appearing in the preceding multiplexed-CGH designs is avoided. The multiplexed polygonal-aperture CGH has a much higher diffraction efficiency than that resulting from either the preceding multiplexed CGH designs or the conventional CGH designed by the IFTA. PMID- 11899269 TI - Self-focusing hidden bar code. AB - A new type of diffractive optical bar code produced by computer-generated holographic technology is proposed. The message in the proposed bar code is hidden in the diffracted light of the bar code element and can be read from the first diffraction order. In contrast to the conventional hidden bar code, which needs a lens to focus the diffracted light, the proposed hidden bar code has a property of self-focusing. This self-focusing ability is achieved by modulating a function of the Fresnel zone plate into the bar code format. Consequently, the read-out process for the information in this hidden bar code avoids the use of a lens. Experiments have shown the feasibility of the proposed bar code and confirmed that it can perform better than the conventional hidden bar code. PMID- 11899271 TI - Principal-component characterization of noise for infrared images. AB - Principal-component decomposition is applied to the analysis of noise for infrared images. It provides a set of eigenimages, the principal components, that represents spatial patterns associated with different types of noise. We provide a method to classify the principal components into processes that explain a given amount of the variance of the images under analysis. Each process can reconstruct the set of data, thus allowing a calculation of the weight of the given process in the total noise. The method is successfully applied to an actual set of infrared images. The extension of the method to images in the visible spectrum is possible and would provide similar results. PMID- 11899270 TI - Characterization of diffraction patterns directly from in-line holograms with the fractional Fourier transform. AB - We show that the fractional Fourier transform is a suitable mechanism with which to analyze the diffraction patterns produced by a one-dimensional object because its intensity distribution is partially described by a linear chirp function. The three-dimensional location and the diameter of a fiber can be determined, provided that the optimal fractional order is selected. The effect of compaction of the intensity distribution in the fractional Fourier domain is discussed. A few experimental results are presented. PMID- 11899272 TI - Photon folding for imaging in nonfocusing telescopes. AB - We present a new technique--photon folding--for imaging in nonfocusing telescopes. Inspired by the epoch-folding method in timing analysis, the photon folding technique directly provides the statistical significance of signals in images, by use of projection matrices. The technique is robust against common imaging problems such as aspect errors and a nonuniform background. The technique provides a deterministic recursive algorithm for improving images, which can be implemented on line. The higher-order photon-folding technique allows a systematic correction for coding noises, which is suitable for studying weak sources in the presence of highly variable strong sources. The technique can be applied to various types of nonfocusing telescopes such as coded-aperture optics, rotational collimators, or Fourier grid telescopes. PMID- 11899273 TI - Reflective grating interferometer: a folded reversal and shearing wave-front interferometer. AB - The reflecting grating interferometer (RGI) is a folded and reversal wave-front interferometer sensitive only to asymmetrical aberrations such as third-order coma. The RGI can isolate and evaluate coma both in nearly collimated and in noncollimated beams. We propose a RGI with a different optical configuration that includes a lateral shearing in addition to folding and reversal operations. With lateral shear, the RGI also becomes sensitive to other terms of third-order aberrations such as defocusing, astigmatism, and spherical aberration. Optical path difference equations for interpreting interferograms and numerical simulations are presented to show how the interferometer works in the shearing configuration. Its potential applications are described and discussed. PMID- 11899274 TI - Three-dimensional optoelectronic stacked processor by use of free-space optical interconnection and three-dimensional VLSI chip stacks. AB - We present a demonstration system under the three-dimensional (3D) optoelectronic stacked processor consortium. The processor combines the advantages of optics in global, high-density, high-speed parallel interconnections with the density and computational power of 3D chip stacks. In particular, a compact and scalable optoelectronic switching system with a high bandwidth is designed. The system consists of three silicon chip stacks, each integrated with a single vertical cavity-surface-emitting-laser-metal-semiconductor-metal detector array and an optical interconnection module. Any input signal at one end stack can be switched through the central crossbar stack to any output channel on the opposite end stack. The crossbar bandwidth is designed to be 256 Gb/s. For the free-space optical interconnection, a novel folded hybrid micro-macro optical system with a concave reflection mirror has been designed. The optics module can provide a high resolution, a large field of view, a high link efficiency, and low optical cross talk. It is also symmetric and modular. Off-the-shelf macro-optical components are used. The concave reflection mirror can significantly improve the image quality and tolerate a large misalignment of the optical components, and it can also compensate for the lateral shift of the chip stacks. Scaling of the macrolens can be used to adjust the interconnection length between the chip stacks or make the system more compact. The components are easy to align, and only passive alignment is required. Optics and electronics are separated until the final assembly step, and the optomechanic module can be removed and replaced. By use of 3D chip stacks, commercially available optical components, and simple passive packaging techniques, it is possible to achieve a high-performance optoelectronic switching system. PMID- 11899275 TI - Measurement of the thermal coefficients of rewritable phase-change optical recording media. AB - We describe a method to estimate the heat capacity of the substrate, the dielectric layer, and the phase-change layer of phase-change optical recording media as well as the thermal conductivity of the phase-change layer in its crystalline state. Measurements were carried out on spinning disks with the beam of light focused and locked onto the groove track. The method relies on the identification of the solid-to-liquid phase transition that occurs in the phase change layer and takes advantage of the dependence of thermal diffusion on track velocity and irradiation time. PMID- 11899276 TI - Optical characterization of multilayer stacks used as phase-change media of optical disk data storage. AB - We report results of measurements of the optical constants of the dielectric layer (ZnS-SiO2), reflecting layer (aluminum-chromium alloy), and phase-change layer (GeSbTe, AgInSbTe) used as the media of phase-change optical recording. The refractive index n and the absorption coefficient k of these materials vary to some extent with the film thickness and with the film deposition environment. We report the observed variations of optical constants among samples of differing structure and among samples fabricated in different laboratories. PMID- 11899277 TI - Polarization dependence of thermal behavior in rewritable phase-change optical recording media. AB - Thermal behavior in land-groove phase-change optical recording has been examined for different polarizations of the incident beam. Three-dimensional temperature distribution in the grooved medium is evaluated by the numerical solution of Maxwell's equations and the heat transfer equation with the finite-difference method. Both experiments and calculations have shown that the thermal behavior in the medium is dependent on the state of polarization and the nature of the track. The calculated mark shapes in a quadrilayer stack are in good agreement with experimental observations. PMID- 11899278 TI - The aging lung: a challenging entity respiratory medicine. PMID- 11899279 TI - Allergenic components of vaccines and avoidance of vaccination-related adverse events. AB - Vaccines have had a dramatic effect on the prevalence of communicable diseases, but, in selected individuals, the injection presents a risk of anaphylaxis. Fortunately, most people have no allergic reactions to vaccines. In egg-allergic individuals, care must be taken before administering specific vaccines; the algorithm provided in this article gives specific recommendations for skin testing and desensitization. This algorithm is not needed for individuals receiving the measles-mumps-rubella vaccine because the risk of anaphylaxis is extremely low, even in those with known egg-protein sensitivity. Some individuals have gelatin sensitivity, which may cause anaphylaxis. Selected vaccines contain antibiotic drugs, so it is important to note if an individual has any known drug sensitivity, especially to neomycin, polymyxin B, or amphotericin B. Lastly, vaccine preservatives may cause reactions, but this occurs very infrequently. PMID- 11899280 TI - Multiple drug allergy syndrome: a distinct clinical entity. AB - Multiple drug allergy syndrome is a clinical condition characterized by a propensity to react against different, chemically unrelated antibiotic or nonantibiotic drugs. The origin of this syndrome is still elusive. This article critically examines the medical literature on multiple drug allergy syndrome, compares and discusses recent personal observations obtained in patients with intolerance to multiple antibiotic or anti-inflammatory drugs, suggests possible pathogenic mechanisms for this type of drug allergy, and reports on current personal research in this field. PMID- 11899281 TI - Structural determinants of antibiotic allergy. AB - Allergies to antibiotics, mainly the beta-lactam antibiotics (penicillins and cephalosporins), are a common, costly, and potentially dangerous clinical problem encountered in everyday practice. Although studies on the role of non-beta-lactam antibiotics in allergic diseases, particularly the development of specific diagnostic tests and the immunochemical identification of allergenic structures, have been too few and relatively superficial, the situation with the beta-lactam antibiotics is much more advanced. Good progress has been made in identifying the spectra of allergenic determinants recognized by IgE antibodies in the sera of subjects sensitized to penicillins and cephalosporins, and this is aiding the development of an appropriate battery of drug conjugates for use as diagnostic agents. Patient-sensitivity responses to the beta-lactam antibiotics are frequently heterogeneous, and this factor must be taken into account for any diagnostic strategy or future therapy with a penicillin or cephalosporin. PMID- 11899282 TI - Latex allergy: review of recent advances. AB - Latex allergy is an IgE-dependent immediate hypersensitivity reaction to latex proteins. Risk factors for latex allergy are contact with latex products and atopy. Children who undergo multiple surgical procedures and healthcare workers are the major groups at risk. Powdered latex gloves are an important source of sensitization. Preventive measures are leading to reduction in latex sensitization and allergic reactions. The prevalence of latex allergy in the general population may be as low as 0.1%, whereas the frequency of latex sensitization is reported to be 7%; this may be due to cross-reacting antipollen IgE. The most important latex allergens have been purified, and some have been cloned and sequenced. Many latex-allergic patients are also allergic to common plant-derived aeroallergens and foods. The structural and biologic relationships among plant-derived food allergens, including latex, explain these clinically important cross-reactions. PMID- 11899283 TI - Atopic eczema and allergy. AB - Although the pathomechanisms of respiratory atopy are well established, the role of IgE-mediated hypersensitivity in the elicitation and maintenance of eczematous skin lesions in atopic eczema (AE) is still controversial. There is, however, evidence for exogenous elicitation of AE by contact with aero- or food allergens (house dust mite, cat, and so forth). Recent investigations show that epidermal Langerhans' cells bind IgE via different receptors, especially the high-affinity receptor (Fc epsilon RI), which is significantly more strongly expressed in lesional skin of AE compared with other inflammatory skin diseases including allergic contact dermatitis. The clinical relevance of IgE-mediated sensitization in AE has been evaluated by the so-called atopy patch test (APT). The APT shows a much higher specificity compared with the skin prick test and radioallergosorbent test. However, allergic reactions do not play a decisive role in every case of AE. Other factors, such as nonspecific skin irritability or psychosomatic interactions, have to be considered. The concept of "extrinsic" versus "intrinsic" types of AE seems attractive. The concept of AE starting with TH2 inflammation, becoming TH1 inflammation in chronicity, and finally progressing to an autoimmune disease with IgE antibodies against autologous epidermal proteins is very attractive. Based on new knowledge, new methods in diagnosis, treatment, and prevention will develop, including more effective avoidance strategies, more potent anti-inflammatory treatment (e.g., immunomodulation or topical immunophyllins), and new ultraviolet modalities. The new findings have given rise to a possible new classification of eczema/dermatitis. The concept of "patient management," including all aspects from avoidance to therapy, has gained acceptance. PMID- 11899285 TI - Immunotherapy for food allergy. AB - Food allergy is an important cause of life-threatening hypersensitivity reactions. Avoidance of allergenic foods is the only method of prevention that currently is available for sensitized patients. This method of prevention is difficult and often impossible. With better characterization of allergens and better understanding of the immunologic mechanism, investigators have developed several therapeutic modalities that potentially are applicable to the treatment and prevention of food allergy. Therapeutic options currently under investigation include peptide immunotherapy, DNA immunization, immunization with immunostimulatory sequences, anti-IgE therapy, and genetic modification of foods. These exciting developments hold promise for the safe and effective treatment and prevention of food allergy in the next several years. PMID- 11899286 TI - Food allergy and the respiratory tract. AB - Previous studies have confirmed that IgE-mediated, food allergy-induced respiratory tract symptoms occur, typically accompanied by cutaneous or gastrointestinal symptoms. The possibility that respiratory tract symptoms are food allergy induced should be considered in patients who have a current or past history of one or more of the following: atopic dermatitis, wheezing (or experiencing anaphylactic symptoms) after ingesting a particular food or foods, and confirmed food allergy. Moreover, the work-up of food allergy in asthma should be considered in patients in whom asthma is poorly controlled despite persistent use of appropriate asthma medications. A definitive diagnosis of food allergy should be based on clinical history, appropriate laboratory testing, and, when indicated, well-controlled oral food challenges. Treatment is based on establishing a safe elimination diet and an emergency plan for managing reactions caused by accidental ingestion. PMID- 11899287 TI - Skin testing and food challenges for the evaluation of food allergy. AB - Skin testing by prick technique has an excellent safety record in the evaluation of food hypersensitivity. Skin prick tests for the common food allergens are excellent tools for identifying those at very low risk of reaction on eating the food but are of variable value in identifying patients who will be positive on challenge. Intradermal skin tests to foods are less safe and appear to add no predictive information. Skin tests to less common food allergens, especially fruits, are less well characterized and may require use of the food item itself as the source of allergen rather than a commercial extract. For a few foods, the CAP system fluorescent enzyme immunoassay (Pharmacia, Peapack, NJ) recently has been shown to have good ability to identify patients at very high probability of reaction on oral challenge. Oral challenge remains the definitive method of demonstrating sensitivity or tolerance to a food. The double-blind, placebo controlled food challenge is the gold standard of diagnosis, but in many situations, simpler open or single-blind challenge procedures may be substituted. With careful, incremental dosing and a low starting dose, oral challenges for food hypersensitivity have an excellent safety record. Skin prick tests are of little value in the evaluation of adverse food reactions not mediated by IgE. Oral challenge is relied upon in this situation for definitive diagnosis, but challenges may be cumbersome if the time course of the presumed reaction is not rapid. PMID- 11899289 TI - Dietary protein enterocolitis. AB - Dietary protein enterocolitis generally presents in the 1st year of life with diarrhea, emesis, and irritability. When there is a delay in diagnosis, persistent exposure to the offending dietary antigen leads to increasing enteric inflammation manifesting as bloody diarrhea, anemia, dehydration, and failure to sustain normal patterns of weight gain and growth. The extent of enteric inflammation may be limited to mild proctitis, pancolitis, or true enterocolitis with esophagitis, gastritis, enteropathy, and colitis. The offending antigen is usually cow's milk protein or soy protein. A significant number of the infants are exclusively breast fed, especially those with proctitis. In older children, a wide variety of dietary proteins have been implicated. The inconsistency between allergists and gastroenterologists in the clinical definition of the syndrome remains a significant problem. To the allergist, the definition is based on clinical criteria, allergy testing, and response to double-blind food challenge, whereas to the gastroenterologist, it is defined by histologic criteria and the response of clinical and histologic manifestations to elimination diets. To further complicate the issue, European studies have emphasized the alterations in enteric permeability noted in both enteropathy and enterocolitis. In an effort to establish a unified approach, the International Life Sciences Institute sponsored a workshop in late 1998, which resulted in a document entitled "Classification of Gastrointestinal Disease of Infants and Children Due to Adverse Immunologic Reactions." PMID- 11899288 TI - Diagnosis and management of food allergy. AB - Food allergy affects 8% of children under 3 years of age and roughly 2% of the adult population. Major targets include cutaneous, gastrointestinal, and respiratory organs. Clinicians must recognize the spectrum of food allergy in order for these patients to be diagnosed accurately and managed. IgE-mediated reactions can progress rapidly, and severe reactions are often associated with refractory bronchoconstriction. IgE-mediated allergic patients should be equipped with education, an emergency action plan, and injectable epinephrine to aggressively treat food-induced reactions and prevent fatalities. PMID- 11899290 TI - Nutrition basics in food allergy. AB - Strict elimination of foods because of food allergies can interfere with good nutrition. It is essential that the diagnosis of a food allergy be proven thoroughly to avoid unnecessary food restrictions. Comprehensive education should show where allergenic foods are found, how to find appropriate substitutes for the eliminated foods and corresponding nutrients, and how to avoid accidental ingestion of allergenic foods. A nutrition assessment is an essential part of the clinical follow-up. Identifying nutrition-related problems early can correct difficult situations and prevent long-term health consequences. PMID- 11899291 TI - The interaction between particulate air pollution and allergens in enhancing allergic and airway responses. AB - Although the detrimental effects of air pollution on human health have been brought widely to public notice, it appears that less attention has been given to the potential role of toxic air pollutants in the induction of allergic conditions such as asthma, rhinoconjunctivitis, and atopic eczema. A number of large epidemiologic studies have shown that people exposed to intense motor vehicle traffic and its associated emissions are at major risk for allergic symptoms, reduced lung function, and increased sensitization to common airborne allergens. Several laboratory-based studies have demonstrated that particulate air pollutants emitted from motor vehicles can induce allergic inflammation, enhance IgE responses, and increase airway hyperresponsiveness, which could provide an underlying mechanism for the increasing prevalence of allergic diseases. This article reviews the evidence that supports the causative link between particulate air pollution and the sharp increase in the prevalence of type I allergies in developed countries. PMID- 11899293 TI - Factors controlling transduction signaling and proliferation of airway smooth muscle. AB - The airway smooth muscle cell is an active participant in the inflammatory response that accompanies asthma. It can interact with the epithelium and inflammatory cells to produce cytokines and cell surface molecule upregulation. Moreover, smooth muscle cells can alter the composition of the extracellular matrix proteins via changes in the production of matrix metalloproteinases and their tissue inhibitors. These properties may contribute to the increase in the amount of airway smooth muscle that characterizes the asthmatic airway wall and the remodeling that underlies the structural changes that lead to persistent asthma. PMID- 11899292 TI - Airway remodeling in the pathogenesis of asthma. AB - Asthma is characterized by a chronic inflammatory process of the airways followed by healing, the end result of which is an altered structure referred to as airway remodeling. Although the mechanisms responsible for such structural alterations appear to be heterogeneous, it is likely that abnormal airway cell dedifferentiation, migration, and redifferentiation, together with changes in connective tissue deposition, contribute to the altered restitution of airway structure and function. This altered restitution is often seen as fibrosis and increased smooth muscle, mucus gland mass, and vessel area. As a consequence of these structural changes, the airway wall in asthma is usually characterized by increased thickness and markedly and permanently reduced airway caliber. These features may result in increased airflow resistance, particularly when there is bronchial contraction and bronchial hyperresponsiveness. The effect on airflow is compounded by increased mucus secretion and inflammatory exudate, which not only block the airway passages but also cause increased surface tension favoring airway closure. PMID- 11899295 TI - The bronchial epithelium in chronic and severe asthma. AB - Although patients with severe, steroid-refractory asthma represent a minor proportion of the asthmatic population, they consume a disproportionate amount of healthcare costs and have a greatly impaired quality of life. They respond poorly to conventional anti-inflammatory therapy and frequently exhibit a component of fixed airflow obstruction that has been linked to airway wall remodeling. In addition to its classic barrier function, the bronchial epithelium responds to changes in the external environment by secreting cytoprotective molecules and mediators that signal to cells of the immune system. In asthma, the bronchial epithelium is stressed and damaged, with shedding of the columnar cells into the airway lumen. This damage and ensuing repair responses are proposed to orchestrate airway inflammation and remodeling via activation of myofibroblasts in the underlying lamina reticularis. This allows the two cell types to work as a trophic unit, propagating and amplifying the response at the cell surface into the submucosa. Because wound healing involves inflammation, repair, and remodeling processes, this review considers the evidence that exaggerated inflammation and remodeling of the airways arise as a consequence of abnormal injury and repair responses coordinated by the bronchial epithelium, highlighting, where possible, steroid-insensitive components. PMID- 11899294 TI - Effects of inhaled corticosteroids on growth in asthmatic children. AB - This article presents a brief review of the effects of inhaled corticosteroids on growth in children with asthma. All currently available inhaled corticosteroids, where there is adequate data, have been shown to cause significant growth suppression in children in a dose-dependent manner. It is now apparent that there are differences in the growth-suppressive effects of different corticosteroids. Recent evidence confirms that the growth-suppressive effects are short lived and that, at conventional doses, inhaled corticosteroids do not affect final attained adult height. PMID- 11899296 TI - The role of neuroeffector mechanisms in the pathogenesis of asthma. AB - Neural regulation of the airways consists of cholinergic excitatory, adrenergic inhibitory nerves and nonadrenergic, noncholinergic (NANC) nerves. NANC nerves can be either inhibitory or excitatory. Cholinergic nerves form the predominant bronchoconstrictor neural pathway in human airways. Acetylcholine controls neuronal and nonneuronal target cells via a short-lived action at nicotinic and muscarinic receptors. The most important control over acetylcholine release from postganglionic cholinergic nerves is exerted by acetylcholine itself. The M2 autoreceptor is located prejunctionally on postganglionic nerves. Its stimulation limits the further release of acetylcholine. A loss of function in the neuronal muscarinic M2 autoreceptor occurs after exposure to allergen, ozone, or viruses. In human airways, inhibitory NANC (i-NANC) mechanisms are the only neural bronchodilatory mechanisms. The presumed neurotransmitters of the i-NANC system are vasoactive intestinal peptide and nitric oxide. Substance P and neurokinin A have been implicated as the neurotransmitters mediating the excitatory part of the NANC nervous system. NK2 receptors are present on smooth muscle of both large and small airways and mediate part of the bronchoconstrictor effect of tachykinins. Most of the proinflammatory effects of substance P are mediated by the NK1 receptor. Tachykinin receptor antagonists are currently being developed as a possible anti-asthma treatment. An extensive cross-talk exists between nerves and the immune system. The complexity of the picture has increased further as it has become clear that classical neurotransmitters, such as acetylcholine and neuropeptides, are produced by nonneuronal cells. PMID- 11899297 TI - The role of viruses in the induction and progression of asthma. AB - Viral respiratory infections have been related to asthma in several ways. It is well established that viral common colds precipitate exacerbations of asthma. Severe bronchiolitis in early life is related to subsequent wheezing and therefore may represent a marker of susceptibility to asthma; alternatively, it could be involved in the initiation of the disease. Finally, it is possible that some infections may protect from the development of asthma and allergies by promoting a type-1 host response. However, whether respiratory or other viruses could mediate such a protective effect is debated. The design and implementation of novel anti- or proviral strategies targeting asthma depends on the resolution of these questions. This review presents current evidence on the epidemiologic correlations and proposed mechanisms for the involvement of viral infections in the development and progression of asthma. PMID- 11899301 TI - Asthma, atopy, and IgE: what is the link? PMID- 11899300 TI - Susceptibility genes in asthma and allergy. AB - Genome-wide screens for asthma and atopy susceptibility loci have been completed in six population samples. Despite the extensive clinical heterogeneity associated with these phenotypes, 20 chromosomal regions show evidence of linkage (P < 0.01) in three or more population samples. Thus, this survey suggests that at least 20 independent loci influence susceptibility to asthma, atopy, or associated phenotypes and indicates that the genetics of these phenotypes are truly complex. Ongoing studies are aimed at identifying the specific gene or genes in these regions that confer susceptibility to asthma or atopy. PMID- 11899302 TI - Literature alert. PMID- 11899298 TI - Regulation of allergic airways inflammation by cytokines and glucocorticoids. AB - Cytokines mediate the allergic inflammatory response of the airways, and glucocorticosteroids ameliorate allergy symptoms by regulating cytokine expression. Recent studies provide insight into the manner by which cytokines work together to mediate allergic airway disease. Real progress has also been gained in our understanding of subcellular mechanisms of allergic inflammation, particularly the role of transcription factors in regulating the expression of specific cytokine profiles and the differentiation of the TH2 subset. This article provides an update of recently reported findings in this field and highlights emerging concepts of allergic inflammation. PMID- 11899299 TI - Novel drugs for treating asthma. AB - The health burden of asthma is increasing globally at an alarming rate, providing a strong impetus for the development of new therapeutics, particularly drugs that may prevent development of the disease. Currently available inhaled bronchodilators and anti-inflammatory drugs are effective in most asthmatic patients, but this palliative therapy requires long-term daily administration. Despite considerable efforts by the pharmaceutical industry, it has been difficult to develop novel therapeutic agents, the leukotriene antagonists being the only new class of asthma treatments to be licensed in the past 30 years. It is clearly important to understand more about the underlying mechanisms of asthma and about how currently used drugs work before rational improvements in therapy can be expected. There are numerous therapies in clinical development that combat the inflammation found in asthma, specifically targeting eosinophils, IgE, adhesion molecules, cytokines (interleukin-4, -5, -13) and chemokines, inflammatory mediators, and cell signaling (kinase inhibitors). In particular, there is the obvious need for new therapy for severe asthma that is poorly controlled by high-dose corticosteroids as well as agents to counter acute emergency asthma. A long-term goal is to develop disease-modifying immunotherapy that could be introduced in childhood to alter the natural history of asthma. Thanks to the extensive efforts of the pharmaceutical industry, we can expect the introduction of a range of novel therapies for asthma in the near future. PMID- 11899303 TI - Restraint reduction in orthopedics. PMID- 11899304 TI - Thoracoscopic spine surgery: current indications and techniques. AB - The first report of thoracoscopic surgery was in 1910, after Jacobaeus used thoracoscopy to lyse tuberculous lung adhesions. However, it was not until the end of the century that Lewis (1991) recognized the value of thoracoscopic surgery, and Mack (1993) reported the application of video-assisted thoracic surgery (VATS) for spine surgery. VATS is still in its infancy and the application of this technology for spine surgery continues to rapidly expand. The current indications for thoracoscopic spine surgery include tissue biopsies, thoracic paravertebral abscess drainage and debridement, thoracic disc herniation excisions, anterior spinal release and/or fusion for spinal deformity, stabilization and fusion of thoracic and thoracolumbar fractures, corpectomy for vertebral tumors, and the placement of anterior spinal instrumentation with fusion. This article reviews these current indications for VATS--the technique and subsequent nursing implications. PMID- 11899305 TI - A practical approach to stress fractures. AB - Stress fractures may be fatigue or insufficiency related. Fatigue stress fractures result when healthy bones are exposed to intense and/or repetitive loads for which the bone is not prepared. Insufficiency stress fractures result from normal loads to bones weakened by genetic, metabolic, nutritional, or endocrine processes. Fracture usually begins as a small cortical infarction that progresses as stress increases or continues. Pain is the hallmark symptom of both types. When confronted with signs and symptoms consistent with stress fractures, providers must consider risk factors, comorbid conditions, and whether or not the mechanism of the injury is consistent with the clinical picture. Though the two types of stress fractures are treated differently, in both, the prognoses are dependent on early identification and intervention. PMID- 11899306 TI - Developing a special needs restraint loaner program for transporting medically fragile children. AB - Pediatric orthopaedic patients require special consideration for their transportation needs. A hospital-based program to fulfill these needs promotes patient safety and allows accessibility of needed devices to elective and nonelective patients. Nurses can be an integral part of this process. This article presents the process of establishing a special needs restraint program including the legalities, acquisition of equipment, and training of appropriate personnel. PMID- 11899308 TI - Vancomycin intermediate-resistant Staphylococcus aureus (VISA). AB - Methicillin-resistant Staphylococcus aureus (MRSA) has been an infectious disease problem since the early 1980s and until recently was uniformly susceptible to vancomycin, the drug known as the "last resort." Recent reports indicate that Staphylococcus aureus has continued to mutate and has developed intermediate resistance to vancomycin (VISA). This article lists some of the potential clinical manifestations of Staphylococcus aureus as well as a possible explanation of the phenomenon of antibiotic resistance. The reported cases of VISA are reviewed, and intervention strategies for prevention and control are discussed. PMID- 11899307 TI - Pain and hip fracture outcomes for older adults. AB - PURPOSE: To determine if postoperative pain is a predictor of functional outcomes for elderly hip fracture patients who were previously independent ambulators (with or without assistive device). DESIGN: Prospective comparative survey design. SAMPLE: Convenience sample. 85 hip fracture patients age 65 years or older from two Midwestern urban hospital orthopaedic units. METHODS: Subjects were interviewed between day 2 and 5 (M = 2.6) postoperatively and again 2 months postoperatively. Independent variables of cognitive status and pain status were measured using the Folstein Mini Mental State Exam (MMSE) and two pain measures, the Verbal Descriptor Scale (VDS) and Ferrell's Pain Experience Interview (FPEI). The dependent variable, functional outcome, was measured using the degree of assistance required for basic ADLs from Jette's Functional Status Index (FSI). FINDINGS: Pain with movement was significantly higher than pain at rest (p < .0001). Mental status, pain report with movement (during hospital interview), illness severity, and age accounted for 51% of the variance in functional outcomes 2 months postoperatively. CONCLUSIONS: Undertreated postoperative pain contributes to poor functional outcomes. IMPLICATIONS FOR NURSING PRACTICE: Pain assessment of postoperative older patients should be conducted during movement. Efforts to reduce postoperative pain severity in the immediate postoperative period may yield better functional outcomes months later. PMID- 11899309 TI - Acute care for elders (ACE): a holistic model for geriatric orthopaedic nursing care. AB - The normal aging process brings about inevitable and irreversible changes in physical, psychosocial, and spiritual health. These normal changes are partially responsible for the increased risk of developing health-related problems in the hospitalized elder population. The Acute Care for Elders (ACE) model provides an effective, proactive, inexpensive framework for addressing the complex health needs of older adults. When ACE principles are used for the clinical management of clients with orthopaedic problems, interdisciplinary conferences provide the structure for maintaining the continuity of care. Mobility as well as independent functioning, comfort level, mental status, depression, skin health, nutrition, and response to treatment, are discussed and nurse initiated guidelines for preventive and restorative interventions are implemented. Follow-up phone calls and/or home visits are important indices of thorough discharge planning. The theory of comfort is used to assure that holistic needs are addressed. Nurses who practice the ACE model are excited about demonstrating the highest level of competency in geriatric nursing, whereby patient functioning is maximized, comfort and dignity are promoted, functional decline is prevented, and patients are successfully returned to their homes. PMID- 11899310 TI - Research utilization in nursing: the power of one. AB - Common barriers to research utilization in nursing include characteristics of the setting in which nurses practice, nurses themselves, and nursing's dependence on rituals and traditions in practice. Nurses can overcome these barriers by questioning their practice and adopting attitudes and values that prioritize research utilization. The "Power of One" Model of Research Utilization guides nurses to examine everyday practices, assess their research foundations, and implement and evaluate changes to research-based practice. PMID- 11899311 TI - Deep muscle contusion complicated by myositis ossificans (a.k.a. heterotopic bone). AB - One muscle trauma is a common injury in contact sports. The most extensively damaged muscle is that segment closest to the underlying bone. By an unknown mechanism, this injured muscle can become transformed into heterotopic bone, a complication also called myositis ossificans. It is best managed conservately and rarely requires surgery. PMID- 11899312 TI - When the family asks, 'what happened?'. AB - Because of the high level of acuity of hospitalized patients, untoward events can and do occur. Very often, nurses develop a caring relationship with the families of these patients. As a result, the family may approach the nurse about this negative turn of events. The questions that the family raises may create an ethical dilemma for the nurse. The nurse may wonder how to respond, feel powerless and "caught in the middle," and experience moral distress because of constraints in the health care system. This article discusses the ethical perspective of caring and the "nurse in the middle" phenomenon. Several strategies to help nurses manage this issue include consulting with a mentor, consulting with the institutional ethics committee, and promoting an ethical climate within the health care setting. PMID- 11899313 TI - Revisiting NAON Foundation 101. PMID- 11899314 TI - Nursing and health care issues in the 2001 provincial election. PMID- 11899315 TI - RNs explore innovative approaches to health care delivery. PMID- 11899317 TI - Nursing news from health Canada. PMID- 11899318 TI - Finest hour (reflections of an OR nurse). PMID- 11899319 TI - The RNFA--anesthetic team collaborative connection. AB - A requirement of the Registered Nurse First Assistant (RNFA) Clinical Internship is to explore the RNFA collaborative role with the anaesthetic team in facilitating positive patient surgical outcomes. PMID- 11899316 TI - Ethics in the workplace. PMID- 11899320 TI - At war for perioperative nurses. PMID- 11899321 TI - Managing conflict to gets your needs met. PMID- 11899322 TI - Epidural cooling for spinal cord protection during thoracoabdominal aortic aneurysm repair (a case study). AB - Aneurysms result from damage to artery walls as a result of underlying athrosclerotic and/or thromboembolic disorders. A thoracoabdominal aortic aneurysm involves vessel damage and wall weakening in the thoracic and abdominal segments of the aorta. Thoracoabdominal aortic aneurysm repair is considered to be high risk due to the nature of the intervention that requires an extensive incision with clamping of the thoracic aorta above the renal arteries. Clamping of the aorta renders all areas distal to the clamp at high risk for ischemic trauma especially to the spinal cord where the risk of neurological deficits postoperatively is 7-16% (Cambria, et al., 1997; Davison, et al., 1997). Several adjunct interventions have been tried to reduce the risk of spinal cord injury associated with the ischemia of cross clamping. Epidural cooling has been successful as an adjunct in reducing the neurological deficits. A preoperative nursing assessment indicating the appropriate nursing diagnoses and nursing care required for this patient, allowed for individualization of the plan needed to include this new procedure and plan for best patient outcomes and practices. PMID- 11899323 TI - Cutaneous paraneoplastic syndromes. AB - Paraneoplastic syndromes are caused by hormones or other substances produced by cancer cells and may be the first sign of cancer. A wide range of paraneoplastic syndromes, including endocrine, neurologic, and cutaneous disorders, occurs in patients with cancer. More than 30 cutaneous paraneoplastic syndromes have been identified; this article reviews some of the more common syndromes--acanthosis nigricans, Paget's Disease, acquired ichthyosis, telangiectasia, hypertrichosis lanuginosa acquisita, erythroderma, Bazex's Syndrome, and necrotizing migratory erythema. When these syndromes are diagnosed during the course of a malignancy, professional caregivers may misinterpret them as indicative of metastatic disease or other disorders and patients may be misdiagnosed and not receive optimal treatment. Paraneoplastic syndromes also compromise quality of life by often causing skin impairment and discomfort. Therefore, nurses must be aware of the signs and symptoms of these cutaneous disorders and know how to care for patients with paraneoplastic syndromes. PMID- 11899324 TI - Management of chemotherapy-induced stomatitis. AB - The past decade has brought about major advances in the medical management of cancer. Despite these advancements, significant toxicities often accompany the potential benefits of chemotherapy. One of the most common toxicities associated with chemotherapy administration is the development of stomatitis. Stomatitis is estimated to occur in 40% of all patients undergoing chemotherapy, and its incidence is two to three times higher in patients with hematologic malignancies and those undergoing bone marrow transplant. Many inconsistencies currently exist in strategies to prevent, assess, and treat stomatitis. Unresolved or undiagnosed stomatitis can lead to major complications such as poor treatment outcomes, increased cost of care, diminished quality of life, and, eventually, mortality. Oncology nurses share responsibility in improving patient outcomes related to stomatitis by remaining knowledgeable, using evidence-based practice, and ensuring follow-up. PMID- 11899325 TI - Prognostic information in breast cancer care: helping patients utilize important information. AB - The Oncology Nursing Society's (ONS's) position on quality cancer care states that "quality cancer care incorporates the individual with cancer (and the family) as fully informed partners and decision makers" (ONS, 1997). Patients diagnosed with breast cancer are inundated with information, and oncology nurses help these patients receive quality cancer care by providing and explaining information related to their diagnosis and treatment. This information allows patients to participate in meaningful collaborative decision making. Prognostic tumor markers have provided information that can determine the natural history of breast cancer, identify women with high-risk or aggressive tumors, and help to establish a disease prognosis. PMID- 11899326 TI - Mylotarg approved for patients with CD33+ acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is the most common adult leukemia and has historically been treated with intensive multiagent chemotherapy. In May 2000, the U.S. Food and Drug Administration approved a new agent, gemtuzumab ozogamicin (Mylotarg) to treat patients who are 60 years and older in first relapse with CD33+ AML and not considered candidates for chemotherapy. Gemtuzumab is an antibody-targeted agent that binds specifically to the CD33 antigen that is found on the surface of more than 80% of patients with AML. The agent is administered via i.v. over two hours, and premedication with acetaminophen and diphenhydramine is recommended. Side effects include fever, chills, neutropenia and thrombocytopenia, and asymptomatic hypotension. Clinical remissions have been observed with gemtuzumab, and additional trials with this new agent currently are being conducted. PMID- 11899327 TI - Pharmacologic and biologic therapies in cancer care: Part II. AB - Brigden (1995) described patients with cancer seeking alternative therapies as being on a quest for natural, less toxic therapies. Hydrazine sulfate exemplifies these pursuits. Hydrazine sulfate is relatively inexpensive, easily administered, and reportedly has low toxicity. Lerner (1997) stated that "although Gold has not received the credit that is his due, by staying within the cancer establishment and playing by the rules, he has brought hydrazine sulfate significantly further than Burzynski has brought antineoplastons" (p. 440). PMID- 11899328 TI - Factors affecting end-stage disease quality of life. Patient and caregiver burdens of terminal illness. PMID- 11899330 TI - Bowel obstruction. PMID- 11899329 TI - Denileukin diftitox. AB - Primary cutaneous T-cell lymphomas (CTCLs) encompass a wide variety of lymphomas that are characterized by the localization of the malignant lymphocytes to the skin at presentation. They are slow-growing and rare, occurring in fewer than 1,000 people annually. Patients may go for months to years with skin abnormalities before being diagnosed. Mycosis fungoides and Sezary syndrome are the most common forms of CTCL and are considered to be indolent diseases. Patients with T1 disease have a normal life expectancy, whereas patients who undergo transformation to large cell lymphoma (8%-23% of patients) have a poor prognosis, with mean survival ranging from 2-19 months. PMID- 11899331 TI - Telephone triage. AB - Nurses frequently provide advice and education over the phone. Telephone triage is an area of potential litigation for the nurse. Information must be clear, concise, and accurate. The nurse needs to ascertain that the patient understands the instructions provided and feels free to ask questions and receive clarification of any information that is confusing. Documentation needs to describe the nursing process of data collection, planning, intervention, and evaluation. In this way, patients will receive quality information that allows them to manage symptoms at home, understand when medical interventions are necessary, and avoid unnecessary delays in care. Telephone triage is an integral part of oncology care that covers a broad range of activities, including symptom management, medication renewal, coordination of care, education, and psychosocial support. As telephone triage becomes more "high risk, high volume," nurses must review the triage process used in their clinical settings. Areas to discuss and delineate include differentiation between medical, nursing, and secretarial responsibilities; technical support systems and access; documentation guidelines; telephone triage protocols; staff education; and patient satisfaction (Nauright, Moneyham, & Williamson, 1999). PMID- 11899333 TI - Inquest recommends whistle blowing legislation to protect nurses when reporting problems. PMID- 11899332 TI - Forensic perioperative nursing. Advocates for justice. AB - Facts and evidence have been negated or lost by the inexperience of health care professionals who are not cognizant of the legal requirements concerning potential criminal cases. In the perioperative setting, policy and procedure should provide guidelines for potential criminal cases based on the key concepts and principles of forensic science. Potential forensic cases and traumatic injuries are not limited to major health care centres. All hospitals should have policies and procedures which outline: traumatic injuries/death, staff responsibilities, details of collecting evidence, documentation, chain of custody. The procedure should also include care of victims, suspected perpetrators as well as family/persons accompanying patient. PMID- 11899334 TI - Surgical drainage devices. Improved securement = improved outcomes. AB - The use of securement devices to prevent the risks, discomfort and expense of accidental dislodgment of surgical drains was investigated by the Royal Columbian Hospital after nursing staff began reporting a previously unrecognized post operative pain syndrome. Nursing staff discovered the cause and eliminated the recurring syndrome by taking two preventative measures: securing urinary catheters with a Statlock securement device, and hanging the catheter bags to the OR stable. Both practice changes eliminated traction on the catheter, preventing its migration into the patient's bladder neck that caused the syndrome's symptoms of bladder pain and spasm when the patient woke up. There were modifications to the chest tube securement protocol that also included a Statlock securement device that improved the skin integrity of patients and increased nursing application efficiencies by 100%. PMID- 11899335 TI - Surgical day care predicaments. An overview of the jams & pickles. PMID- 11899336 TI - Unclean scissors implicated in nosocomial infections: nurses' scissors having the most organisms. PMID- 11899338 TI - Peace, the environment, and midwifery: kindred concerns. PMID- 11899337 TI - The future of nursing in the operating rooms of Canada. Are you in the front car of the perioperative train? PMID- 11899339 TI - Midwifery in Ontario today: a reason to celebrate. PMID- 11899340 TI - Ask the midwife. Prevention and care of hemorrhoids, including homeopathic remedies. PMID- 11899342 TI - Vicki's hot pack creations. PMID- 11899341 TI - The vitamin K controversy. PMID- 11899343 TI - Citizens for Midwifery, Inc. National advocates for the midwifery model of care. PMID- 11899344 TI - A legislative conference for midwifery advocates: a participant reports. PMID- 11899345 TI - Toward the end of unnecessary cesarean sections: how the position of your unborn baby affects labor and birth. PMID- 11899346 TI - A new VBAC concern. PMID- 11899347 TI - Another tool in the midwife's bag: transforming professional encounters into an expression of intuitive healing through writing. PMID- 11899348 TI - Two sides to the Cytotec debate. PMID- 11899349 TI - A tale of two cities: Oxford and Bethesda. PMID- 11899350 TI - New horizons in treating metastatic disease. AB - The management of metastatic breast cancer is changing as a consequence of extraordinary discoveries in cancer research and the development of more advanced diagnostic technologies. Although traditional chemotherapeutics such as anthracyclines and taxanes still represent the mainstay of treatment for this disease, new drugs are demonstrating significant clinical activity and sometimes a better toxicity profile. Furthermore, the successful introduction into clinical practice of biological agents, in particular the monoclonal antibody trastuzumab, offers a key to the future of managing metastatic breast cancer. A therapeutic approach based on modifications of a specific molecular target (e.g., gene therapy, vaccines, and antiangiogenesis) alone or combined with the traditional chemotherapeutic drugs is expected to be used more commonly and will, we hope, bring significant improvement in the clinical response and quality of care of our patients. PMID- 11899351 TI - Capecitabine in breast cancer: current status. AB - Anthracyclines, together with taxanes, are at present the most active agents in metastatic breast cancer, while single-agent, bolus 5-fluorouracil (5-FU) is not very active in this setting. In view of encouraging results and tolerable toxicity of continuous infusion of 5-FU in gastrointestinal cancer, innovative oral 5-FU agents such as capecitabine have been developed. Capecitabine is a prodrug that is converted into the active compound 5-FU preferentially at the tumor site. An intermittent dosing schedule of capecitabine twice daily at a dose of 2510 mg/m2/day on days 1-14 in a 3-week cycle appeared to be feasible and resulted in a high dose intensity. A large phase II study investigating capecitabine in 135 advanced breast cancer patients, pretreated with anthracyclines and taxanes, observed three complete and 24 partial responses (response rate, 20%), with a mean duration of 8.0 months. Preliminary results of a study comparing capecitabine with paclitaxel in 42 breast cancer patients failing anthracyclines indicate that the efficacy of capecitabine is comparable to that of paclitaxel, with response rates of 36% and 21%, respectively. Another study reported a response rate of 25% for capecitabine as first-line therapy for advanced breast cancer in women aged > or = 55 years, which tended to be better than combination chemotherapy with cyclophosphamide/methotrexate/5-FU. In all studies, capecitabine side effects were mainly mild, and treatment-related grade 3/4 toxicity consisted of diarrhea (8%-11%), nausea (4%-11%), hand-foot syndrome (10%-18%), neutropenia (3%-20%), and bilirubin elevation (6%). Capecitabine is clearly an active agent for the treatment of breast cancer. It is currently registered in various countries for use in third-line treatment of metastatic disease. Its further role will have to be defined from data of randomized phase III studies. PMID- 11899352 TI - Resistance to endocrine therapy of breast cancer: recent advances and tomorrow's challenges. AB - The role of endocrine therapy in early as well as advanced breast cancer cannot be overrated. Long-term tamoxifen exposure (5 years) in the adjuvant setting has been shown to be effective not only in improving relapse-free and overall survival but also in reducing the incidence of contralateral cancers. Promising results have been achieved in breast cancer prevention with use of antiestrogens. Novel aromatase inhibitors and inactivators have been found superior to conventional treatment in metastatic disease and are currently being evaluated in the adjuvant setting to improve relapse-free and overall survival. If potential health hazards from estrogen deprivation with regard to cardiovascular disease as well as bone metabolism can be addressed, adjuvant endocrine therapy may include such drugs in the future. However, while endocrine therapy of breast cancer has become more and more important in the clinic, the major problems in hormonal therapy are primary and acquired resistance to endocrine manipulations. The causes for endocrine resistance and possible ways to delay or avoid this phenomenon are only allusively understood. Elucidation of the mechanisms underlying endocrine resistance in vivo represents the key to improve our treatment strategies. Due to intense use of in vitro models and animal systems, many potential mechanisms of endocrine resistance have been described; however, our understanding of the problem of drug resistance in vivo remains limited. Hopefully, ongoing programs on translational research in the neoadjuvant, adjuvant, and palliative settings will provide information that will improve our understanding of the biology of endocrine resistance in vivo and, thus, provide us with a better rationale to improve early as well as late endocrine therapy in breast cancer patients. The present publication summarizes the state of the art with respect to endocrine resistance. PMID- 11899353 TI - Low proliferative rate of invasive node-negative breast cancer predicts for a favorable outcome: a prospective evaluation of 669 patients. AB - This study was designed to compare outcome in terms of disease-free survival (DFS) in women with histologically negative axillary lymph nodes and documented low proliferative rate cancer to other well-defined prognostic factors including type of adjuvant treatment. Between 1988 and 1998, we studied 669 patients with invasive node-negative breast cancer up to 5 cm in size and low proliferative rate measured by flow cytometry to determine S-phase fraction (SPF) or by histochemistry (Ki67/MIB1). At a median follow-up of 53 months, 5-year DFS for the entire group was 94% and did not differ significantly by type of systemic adjuvant treatment: none (133 patients, 95% DFS), tamoxifen (441 patients, 94% DFS), or chemotherapy with doxorubicin and cyclophosphamide (95 patients, 92% DFS). In a multivariate prognostic factor analysis, only tumor size was significant; 5-year DFS was 96% for T1N0 cancer versus 89% for T2N0 cancer (P = 0.01). We have prospectively confirmed that a low rate of proliferation as measured by SPF or MIB1 determination confers an excellent prognosis in invasive node-negative breast cancer up to 5 cm in size, regardless of adjuvant treatment. PMID- 11899354 TI - Ocular metastases from breast cancer. AB - Breast cancer is the most common cancer to metastasize to the eye. This is thought to be a rare occurrence but may be more common than thought. Two cases with eye metastases from breast cancer are presented to acquaint physicians with this entity. Other tumors that metastasize to the eye, the anatomic location of metastases in the eye, and the evaluation, treatment, and prognosis of breast cancer metastatic to the eye are discussed. PMID- 11899355 TI - Hormones 'R' us. PMID- 11899358 TI - Selective estrogen receptor modulators as a new therapeutic drug group: concept to reality in a decade. AB - This article provides an overview of the historical development, current research, clinical benefits, and potential future applications of the selective estrogen receptor modulators (SERMs), tamoxifen and raloxifene. The understanding of the mechanism of action of SERMs led not only to the development of tamoxifen, the first widely used antiestrogen for breast cancer treatment, but also to its application as a chemopreventive agent. The SERM principle of antiestrogenic actions in the breast but estrogenlike actions in bone is reviewed in clinical practice through analysis of the current applications and the potential for expanding the role of SERMs. The current view of the molecular mechanism of SERM action is summarized to identify potential target sites for future research. The clinical success of tamoxifen and raloxifene for the prevention and treatment of breast cancer and osteoporosis, respectively, has encouraged the development of a range of new agents that target breast cancer, osteoporosis, coronary heart disease, and endometrial safety. PMID- 11899359 TI - A rationale for the reinitiation of adjuvant tamoxifen therapy in women receiving fewer than 5 years of therapy. AB - The overview analysis of adjuvant therapy trials in early-stage breast cancer by the Early Breast Cancer Trialists' Collaborative Group (EBCTCG) compared 1, 2, and 5 years of adjuvant therapy. The optimal benefit to patients in terms of reduced risk of recurrence and death was clearly conferred by 5 years of therapy with tamoxifen compared to shorter durations of therapy. Many women with early stage breast cancer treated in the past received a recommendation for shorter durations of tamoxifen therapy. The obvious question is whether reinitiating tamoxifen to complete a full 5-year course would further reduce the risk of recurrence and death. In an attempt to explore this issue, we have analyzed recurrence rates in women who received different durations of tamoxifen therapy in the adjuvant setting. The data used for this analysis are directly from the EBCTCG. Our analysis suggests that women who received 1-2 years of adjuvant tamoxifen therapy may benefit from reinitiation of tamoxifen to complete a 5-year course. PMID- 11899360 TI - Taxanes and capecitabine in combination: rationale and clinical results. AB - The clinical utility of capecitabine as a single agent in metastatic breast cancer has been demonstrated with significant responses seen in women already treated with anthracyclines and taxanes. A phase II study in older women with metastatic breast cancer demonstrated capecitabine to be an effective front-line therapeutic agent. Clinical trials of capecitabine in combination with the taxanes, paclitaxel and docetaxel, have been based on the observed upregulation of thymidine phosphorylase (TP) in preclinical studies. This taxane-mediated upregulation is synergistic, time dependent, and persists for up to 10 days. Studies of taxanes administered every 3 weeks with capecitabine have shown favorable antitumor responses and the combination of a taxane with capecitabine was favored over a taxane alone. The day-to-day administration of taxanes and capecitabine led to toxicity concerns, which have hindered their daily use. The administration of taxanes on a weekly schedule has demonstrated a more favorable toxicity profile (i.e., less myelosuppression), and initial studies in combination with capecitabine have demonstrated their utility in various solid tumors. Schedule optimization based on the upregulation of TP may result in a greater therapeutic index, thus allowing for the determination of the most advantageous way of combining these agents. PMID- 11899361 TI - Young age as an adverse prognostic factor in premenopausal women with operable breast cancer. AB - Data regarding young age as an independent prognostic factor have been conflicting. We investigated this variable in 696 premenopausal Vietnamese and Chinese women with operable breast cancer who participated in a clinical trial of adjuvant surgical oophorectomy and tamoxifen. Tumor size and axillary lymph node status did not vary with age. Women < 35 years had a greater fraction of histologic grade III tumors (P = 0.06), and in the two thirds of patients with available data, in women < 35 years, there was a lower percentage of estrogen- and progesterone receptor-positive tumors and a higher percentage of HER2/neu positive tumors (P > 0.14 for each group). In univariate analyses, compared to women > or = 45 years, women < 35 years and 35-39 years were at greater risk for death (P = 0.002 and P = 0.023, respectively), and compared to women > or = 40, women < 40 were at greater risk of death (P = 0.002). Multivariate analyses supported a conclusion that younger age was an independent adverse prognostic factor for survival (P = 0.005, age as a continuous variable). Kaplan-Meier analyses in all patients and in oophorectomy and tamoxifen-treated patients, but not in observation-only patients, showed statistically significant poorer disease free and overall survival in women < 40 years compared to those > or = 40 years. Thus, despite efficacy of the combined adjuvant hormonal therapy, younger age was a risk factor for poorer survival. PMID- 11899362 TI - Phase I/II trial of gemcitabine plus cyclophosphamide in patients with metastatic breast carcinoma previously treated with taxanes. AB - The purpose of this study was to determine the maximum tolerated dose (MTD) of infusional gemcitabine given in conjunction with intravenous (i.v.) cyclophosphamide, and to determine whether the regimen produced a response rate of at least 40% in patients with metastatic breast cancer who have been previously treated with taxanes. Patients received cyclophosphamide (600 mg/m2) i.v. followed immediately by gemcitabine (100, 150, or 200 mg/m2) given as a 24 hour infusion (every 3 weeks) using an accelerated dose-escalation schema. Dose limiting toxicity was defined as a neutrophil nadir < 500/microL, platelet nadir < 50,000/microL, or > or = grade 2 nonhematologic toxicity (> or = grade 3 toxicity during the standard dose-escalation portion of the study). Twelve patients received a total of 32 cycles of therapy. The MTD of gemcitabine was 150 mg/m2. Dose-limiting toxicities at 200 mg/m2 included neutropenia and mucositis. One patient with lymphangitic lung metastases had a partial response (8%; 95% confidence intervals: 0%, 23%). This patient developed grade 4 transaminase and total bilirubin elevation that occurred after the sixth cycle of therapy. The study was terminated due to an insufficient number of responses. The MTD of gemcitabine given as a 24-hour infusion is 150 mg/m2 when used in conjunction with cyclophosphamide (600 mg/m2) every 3 weeks. This regimen is not likely to produce more than a 40% response rate in patients with metastatic breast cancer previously treated with taxanes. PMID- 11899364 TI - Serum interleukin 6, plasma VEGF, serum VEGF, and VEGF platelet load in breast cancer patients. AB - Serum and plasma concentrations of vascular endothelial growth factor (sVEGF and pVEGF), serum concentrations of interleukin 6 (IL-6), and VEGF platelet load (VEGF/pl) in the blood of healthy controls (n = 26), breast cancer patients with locoregional disease (n = 31), and patients with progressive advanced disease (n = 73) have been compared. The 95th percentile values for the control population were 250 pg/mL for sVEGF, 30 pg/mL for pVEGF, and 1.6 pg/mL for IL-6. The 95th percentile value of the calculated VEGF/pl was 1.0 pg/10(6) platelets in the control population. Serum VEGF concentrations correlated with platelet number in all the groups. Patients with thrombocytosis had a median sVEGF concentration of 833 pg/mL, compared to 249 pg/mL in other patients (P = 0.018). Serum IL-6 levels correlated with sVEGF levels and with the calculated VEGF/pl. Serum IL-6 concentration was significantly higher in patients with breast cancer compared to healthy controls (P < 0.0001). Median IL-6 serum levels were nearly 10 times higher in patients with metastatic breast cancer as compared to the those with locoregional disease (6.0 pg/mL versus 0.7 pg/mL, respectively). Plasma VEGF and the VEGF/pl were also significantly different in the 3 groups. The ratio between sVEGF and pVEGF tended to be smaller in the metastatic breast cancer group compared to the patients with locoregional disease (median, 7.5 versus 10.1, respectively; P = 0.066), suggestive of more intravasal platelet degranulation in the former group. Serum IL-6 level is the most discriminative factor separating healthy controls and the locoregional and metastatic breast cancer patient groups. These results suggest a role for tumor-derived IL-6 in regulating VEGF expression in platelets and their precursors and also confirm the role of circulating platelets in the storage of VEGF. PMID- 11899363 TI - Assessment of response by breast helical computed tomography to neoadjuvant chemotherapy in large inflammatory breast cancer. AB - Breast helical computed tomography (CT) was evaluated for use in assessing response to neoadjuvant chemotherapy and residual tumor volume. Forty-three patients with large, inflammatory breast cancers (stage IIA, 12; IIB, 13; IIIA, 9; IIIB, 9), all histologically confirmed by core biopsy, were evaluated prior to and following neoadjuvant chemotherapy. The breast helical CT procedure involved patients in the prone position using single acquisition during quiet respiration following intravenous injection of nonionic contrast material. Helical CT results (3.2-mm slices and maximum intensity projections) were compared to clinical and mammographic evaluations, as well as to pathologic findings. All tumors were clearly visible by breast helical CT, showing important tumor enhancement. Helical CT evaluation of response to chemotherapy (using World Health Organization criteria) corresponded better with mammography (78%, Cohen's kappa statistic (kappa) = 0.65) than with clinical examination (53%, kappa = 0.30). Helical CT measurement of residual tumor volume after neoadjuvant chemotherapy and correlation with pathologic findings were globally satisfactory. The intraclass correlation coefficient was 0.69 (excellent for rounded opacities [0.97], but not as good for diffuse, scattered or multinodular opacities [0.60]). By contrast, clinical and mammographic correlations were globally unsatisfactory (0.49 and 0.28, respectively). Breast helical CT can be very useful in the quantitative assessment of response to neoadjuvant chemotherapy and preoperative determination of residual tumor volume. For this reason, it can be considered an alternative to breast magnetic resonance imaging because of its simplicity, rapidity, and accessibility. PMID- 11899365 TI - Trastuzumab for breast cancer-related carcinomatous meningitis. PMID- 11899366 TI - The impact of tobacco use in women: exploring smoking cessation strategies. AB - The incidence of lung cancer in women has escalated during the last several decades. Lung cancer death rates in women also have risen and now exceed the number of deaths from breast cancer. Tobacco use accounts for more than 30% of all cancer deaths. Currently, 22 million adult women smoke, and more than 1.5 million adolescent females are smokers (American Cancer Society, 2000a). The use of tobacco by young female adolescents is on the rise, and those who are current smokers typically began smoking prior to high school graduation. Oncology nurses have an opportunity in inpatient and outpatient settings to impact the smoking habits of females, regardless of age. This article presents the guidelines for assisting women in smoking cessation. Clinical implications are presented that all oncology nurses should consider implementing in their practice setting. PMID- 11899367 TI - Cancer prevention and early detection. AB - Advances in the area of cancer prevention and early detection are being made constantly. Basic epidemiologic research continues to provide insight into the impact of carcinogen exposure and the development of cancer. It is exciting to note that the study of Tamoxifen and Raloxifene chemoprevention trial is successfully recruiting women, including minority women, to participate. This large chemoprevention trial is providing much insight into how to recruit and retain women to take a chemoprevention agent to ultimately prevent the development of cancer. Advances also are being made in the knowledge base of how to best detect cancer in asymptomatic people. The best screening tool recommendation for the early detection of colorectal cancer remains controversial. Screening for colorectal cancer, however, is the only way to ultimately decrease the morbidity and mortality associated with the disease. Oncology nurses need to accurately risk for colorectal cancer and provide patients with the necessary information to make an informed choice about the most appropriate screening for their situation. Oncology nurses need to be familiar with new research and advances in cancer prevention and early detection so they can share information with patients and their families. PMID- 11899368 TI - Getting ready for JCAHO--just meeting the standards or really improving pain management. AB - The Joint Commission on Accreditation of Healthcare Organizations' revised standards include new standards regarding pain assessment and management. These standards need to be incorporated into patterns of daily practice in healthcare institutions. Documentation systems, policies and procedures, standards of practice, staff education, and quality-improvement programs will need to reflect integration of these standards. Using evidence based consensus statements, guidelines, and other resources on pain management along with a step-wise implementation process will not only enhance success for incorporating the revised standards but also will promote achievement of another goal-optimal assessment and management of pain experienced by other patients with cancer. PMID- 11899369 TI - Unusual port presentation. PMID- 11899371 TI - Hypertension. PMID- 11899370 TI - Factors affecting genetic testing and decisions about prophylactic surgery. AB - Both of the articles reviewed here as well as the references, suggest that very little is actually known about the impact of many aspects of genetic testing. How decision are made about genetic testing in people who do not have cancer, how the results of testing are used used to guide care, and ultimately how people adjust to prophylactic surgery, which is the most effective form of prevention currently available to those who do have a mutation are not completely clear. This has many implications for practice in general. Oncology nurses who build relationships with those diagnosed with cancer and their families may be one of the best groups of professionals to provide the education and counseling individuals and families need prior to making any decision about genetic testing. Just as many responses to cancer exist, so do many responses to finding out the results of mutation status. Oncology nurses are challenged to help facilitate adjustment to learning that one carries a mutation that significantly increases risk of developing cancer. More nursing research needs to be conducted on how to facilitate this adjustment. Dealing with the unknown can be a frightening experience. Little is known about the long-term effectiveness of prophylactic mastectomy and oophorectomy in unaffected mutation-positive individuals. Most of what is known is based on retrospective review. Nurses are challenged to interpret this information, along with its inherent strengths and weaknesses, to individuals so they can make the best possible decisions. The psychosocial needs of those who undergo prophylactic surgery are not clearly understood. Surgery can have many psychological outcomes, and how individuals adjust to these changes is not clear. More nursing research is needed not only to understand these needs but also to design interventions to facilitate and improve adjustment to not only the information that one is mutation positive but also to prophylactic surgery. People who do not have cancer but have a high risk for cancer because of their genetic background need comprehensive and consistent care by knowledgeable healthcare providers. Although these individuals have not been diagnosed with cancer, they have complex psychosocial needs related to their family history and the decisions being made about prevention strategies. Oncology nurses can help fill this gap in care and provide the necessary support these individuals need. PMID- 11899373 TI - Only small percentage of patients choose euthanasia, physician-assisted suicide at end of life. PMID- 11899372 TI - Bend and stretch, reach for the sky. PMID- 11899374 TI - Legislation improves needlestick safety and prevention. PMID- 11899375 TI - Nursing care of patients with multiple myeloma: a paradigm for the needs of special populations. AB - Multiple myeloma, a B cell malignancy that has its peak incidence in the elderly and affects African Americans more frequently than Caucasians, may be used as a paradigm to examine the concerns of special populations. Special populations are those who are medically underserved or medically underrepresented. Using this disease entity to focus on concerns of special populations and access to healthcare systems, the oncology nurse can formulate an approach to cancer care with a broader view of patients from special populations and their needs and goals. PMID- 11899377 TI - A seat at the table. PMID- 11899376 TI - Photodynamic therapy: another option in cancer treatment. AB - Photodynamic therapy (PDT)--also called phototherapy, photoradiation therapy, or photochemotherapy--is a relatively new treatment option being used to treat certain types of cancer, including endobronchial and esophageal cancers. PDT is a two-step process that involves administration of a photosensitizing agent followed by exposure to non thermal laser light. Intensive patient education and support are required during PDT. In the immediate post-procedure period, patients require intensive monitoring to ensure that a patient airway is maintained and pain is alleviated. Photosensitivity persist for four to six weeks post-PDT; to avoid photosensitivity reactions, patients must adhere to specific recommendations to avoid exposure to light. PDT is noted for extending the life expectancy and improving the quality of life of patients. PMID- 11899378 TI - Paclitaxel in the adjuvant setting: results so far are inconclusive. PMID- 11899379 TI - Is HER2/neu overexpression a predictor of anthracycline utility? PMID- 11899380 TI - Results and implications of the Royal Marsden and other tamoxifen chemoprevention trials. AB - A pilot chemoprevention study from the Royal Marsden Hospital in the United Kingdom demonstrated that tamoxifen could be administered safely to healthy women. This led to the establishment of multicenter trials, including the National Surgical Adjuvant Breast and Bowel Project (NSABP) P-1 and Italian National Cancer Institute trials. An interim analysis of the Royal Marsden Hospital trial did not detect a preventive effect. The Italian trial concurred with the Royal Marsden Hospital trial. In contrast, the larger NSABP study detected a 49% reduction in the incidence of breast cancer with tamoxifen chemoprevention. Possible reasons for the different results are examined and the implications for tamoxifen chemoprevention are discussed. PMID- 11899381 TI - Results and implications of the Royal Marsden and other tamoxifen chemoprevention trials: an alternative view. AB - At the time of the release of the findings of the Breast Cancer Prevention Trial (BCPT), early interim results from two smaller European studies were also released. These smaller studies included one from the Royal Marsden (RM) Hospital in London, England, and another from the Italian National Cancer Institute. Since then, there has been much discussion about the relevance of the interim findings from the European studies and the likely reason for the failure of these studies to find a treatment effect. In some instances, the discussion has been incomplete or inconsistent with the observations from the trial. This has caused some confusion regarding the likely differences among the three studies of breast cancer prevention with tamoxifen. Recently, investigators from the RM study have published their interpretation of the reasons for the negative findings from the European studies. The discussions of the RM investigators are reviewed and used as a basis to illustrate some misconceptions regarding key differences in trial design and implementation among the BCPT and the European trials. The investigator discussions are also used to illustrate the significance of performing an appropriate benefit/risk assessment to identify women who would likely have a net beneficial effect when using tamoxifen to reduce the risk of breast cancer occurrence. Differences in terms of the characteristics of the study populations resulting in inadequate statistical power is the most likely reason for the failure to detect treatment differences in the European trials. Possible confounding due to the use of hormone replacement therapy is another reason that must be considered. Also, benefit/risk analysis indicates that tamoxifen has substantial public health potential as an approach to reduce breast cancer incidence and the physical and mental morbidity associated with this disease. The drug cannot be used indiscriminately due to the potential side effects, but benefit/risk assessment methodology can be used to identify substantial numbers of women in whom treatment would provide a net beneficial effect. PMID- 11899382 TI - Activity of pemetrexed (ALIMTA, multitargeted antifolate, LY231514) in metastatic breast cancer patients previously treated with an anthracycline and a taxane: an interim analysis. AB - As many breast cancer patients receive adjuvant chemotherapy using anthracyclines or anthracenediones and taxanes, more therapeutic options are needed for subsequent lines of therapy. Pemetrexed (ALIMTA, multitargeted antifolate, LY231514) is a novel antifolate that inhibits several enzymes in the de novo pathways of pyrimidine and purine biosynthesis. This paper reports on a subset analysis of a phase II clinical trial of pemetrexed in heavily pretreated metastatic breast cancer (MBC) patients. Patients were required to have received prior first-line anthracycline therapy for metastatic disease. Prior adjuvant chemotherapy and prior taxanes were allowed. A substantial subset of the study population (31 of 72 patients, 43%) had also received a taxane in the metastatic setting. All patients were treated with pemetrexed, 600 mg/m2 by intravenous infusion, once every 21 days. In the study subset, 23 of 31 (74%) patients were anthracyclines failures (progression > 30 days following treatment), and eight (26%) patients were anthracyclines refractory (progression during or < or = 30 days of treatment). The median age was 55 years (range, 30-75 years) and the median World Health Organization performance status was 0. Metastases were present in the liver (61%), lung (29%), bone (6%), and soft tissue (19%). The overall response rate for this subset was 26%, with one complete response, seven partial responses, and 13 (42%) patients with stable disease. The median duration of response was 5.4 months and median survival was 12.8 months. Pemetrexed was well tolerated by patients in the study. This post hoc analysis suggests promising activity in MBC patients previously treated with both anthracyclines and taxanes. An ongoing trial is prospectively evaluating activity in this same population. PMID- 11899383 TI - Interim analysis of the use of the anti-idiotype breast cancer vaccine 11D10 (TriAb) in conjunction with autologous stem cell transplantation in patients with metastatic breast cancer. AB - The anti-idiotype monoclonal antibody breast cancer vaccine 11D10 (TriAb) was administered before and after autologous stem cell transplantation (ASCT) in 45 patients with metastatic breast cancer whose disease was responsive to conventional chemotherapy. Evidence of a positive anti-anti-idiotype antibody (Ab3) humoral response was noted at a median of 1.76 months post-ASCT (range, before ASCT-6 months) with this strategy. Maximal Ab3 levels and idiotype specific T-cell proliferative responses were observed at a median of 3 and 4 months, respectively, after ASCT. The achievement of rapid immune responses after ASCT, during a known period of decreased immunoresponsiveness, opens the possibility of an additional antitumor effect at a time when the tumor burden is relatively small. Moreover, in this interim analysis, patients with the most vigorous humoral and cellular immune responses had a significant improvement in progression-free survival. Further follow-up and evaluation of this approach is warranted. PMID- 11899384 TI - Assessment of axillary lymph node involvement in small breast cancer: analysis of 893 cases. AB - Axillary nodal involvement (ANI) remains an essential prognostic factor for breast cancer patients, as it implies the necessity of systemic adjuvant treatment and locoregional irradiation. Axillary dissection (AD) contributes to improved local disease control and may increase survival. However, AD results in a 10%-25% incidence of long-term side effects, particularly lymphedema. Moreover, many small primary lesions with low risk of ANI are now discovered by screening, and it is not clear whether AD should be used routinely in all such patients. Sentinel lymph node biopsy (SLNB) is a selective procedure that allows selective staging of the axilla with few side effects. However, indications for SLNB are not precisely defined yet, so some patients may be understaged and the axillary relapse rate may increase. This study was conducted to help clinicians assess the risk of ANI and analyzed six clinical and histological parameters to optimally recognize patients who might benefit from SLNB, with a minimal risk of false negative rate. We retrospectively analyzed the ANI risk among 893 women treated by conservative surgery and radiation for T0, T1, or T2 invasive tumours < 3 cm in size. All patients underwent AD with sampling of a minimum of seven lymph nodes. In each case, we assessed the clinical and pathological tumor size, histological subtype (including grading), tumor location, age at diagnosis, and breast size. The global ANI rate in the entire cohort was 25.3%. In multivariate analysis, three variables were significantly predictive of the ANI risk: tumor size (P < 0.0001), histological subtype (P = 0.0005), and breast size (P = 0.004). By combining these parameters, we were able to define three categories of women with low (< 20%), intermediate (21%-25%), and high (> 25%) ANI risk. We suggest that women with nonpalpable (T0), T1 grade 1/2, and T2 < 3 cm tumors of medullary, mucinous, tubular, or papillary histological subtype are the best candidates for SLNB. For other patients with a higher ANI risk tumor, AD may still remain the best procedure to obtain accurate staging and definitive local control. PMID- 11899385 TI - Microvessel density and p53 overexpression in young women with breast cancer: a case-control study. AB - Several reports have indicated that young women (less than 40 years of age) with breast cancer have a worse prognosis than older women. We performed a case control study in order to confirm this observation and to determine whether this was attributable to increased microvessel density (MVD) or p53 expression. Twenty six young women (cases) with stage I-III breast cancer that had adequate paraffin embedded archival tissue were identified by the Montefiore Medical Center Tumor Registry over a 24-year period. For each case, two or three control subjects at least 40 years of age or older were selected from the registry and matched for nodal status and tumor size. Immunohistochemistry was performed for MVD and p53 overexpression. A Cox proportional hazard model was performed to examine the influence of age, MVD, p53 overexpression, and recognized prognostic factors on disease-free and overall survival. There were 26 cases (median age, 36 years) and 72 controls (median age, 64 years). The groups were well matched for known prognostic variables. There was no significant difference in p53 overexpression or MVD in the cases and controls. In multivariate analysis, the only features associated with an increased risk of recurrence included young age (hazard ratio [HR] = 2.49; 95% confidence interval [CI]: 1.18-5.25; P = 0.02) and positive lymph nodes (HR = 2.44; 95% CI: 1.12-5.30; P = 0.02). We have confirmed previous reports demonstrating a worse prognosis for women younger than 40 years with invasive breast cancer but found no correlation between young age and MVD or p53 overexpression when adjusted for other variables. PMID- 11899386 TI - Expansion of HER2/neu-specific T cells ex vivo following immunization with a HER2/neu peptide-based vaccine. AB - The identification and characterization of tumor antigens has facilitated the development of immune-based cancer prophylaxis and therapy. Cancer vaccines, like viral vaccines, may be effective in cancer prevention. Adoptive T-cell therapy, in contrast, may be more efficacious for the eradication of existing malignancies. Our group is examining the feasibility of antigen-specific adoptive T-cell therapy for the treatment of established cancer in the HER2/neu model. Transgenic mice overexpressing rat neu in mammary tissue develop malignancy, histologically similar to human HER2/neu-overexpressing breast cancer. These mice can be effectively immunized against a challenge with neu-positive tumor cells. Adoptive transfer of neu-specific T cells into tumor-bearing mice eradicates malignancy. Effective T-cell therapy relies on optimization of the ex vivo expansion of antigen-specific T cells. Two important elements of ex vivo antigen specific T-cell growth that have been identified are (1) the preexisting levels of antigen-specific T cells and (2) the cytokine milieu used during ex vivo expansion of the T cells. Phase I clinical trials of HER2/neu-based peptide vaccination in human cancer patients have demonstrated that increased levels of HER2/neu-specific T-cells can be elicited after active immunization. Initiating cultures with greater numbers of antigen-specific T cells facilitates expansion. In addition, cytokines, such as interleukin-12, when added during ex vivo culturing along with interleukin-2 can selectively expand antigen-specific T cells. Interleukin-12 also enhances antigen-specific functional measurements such as interferon-gamma and tumor necrosis factor-alpha release. Refinements in ex vivo expansion techniques may greatly improve the feasibility of tumor-antigen T cell-based therapy for the treatment of advanced-stage HER2/neu-overexpressing breast malignancy. PMID- 11899388 TI - Taxanes for breast cancer: an evidence-based review of randomized phase II and phase III trials. AB - The taxanes paclitaxel and docetaxel have an important role in the treatment of breast cancer, and numerous randomized trials have evaluated their efficacy for this indication. A systematic, evidence-based review was performed, which included all randomized, controlled trials evaluating taxanes for the treatment of early-or advanced-stage breast cancer that were identified in CANCERLIT and MEDLINE searches. The primary objectives of this review were to determine the dose and schedule for each taxane that was associated with the most favorable therapeutic index, and to determine whether (and under what circumstances) the taxanes improved survival. The search revealed 18 randomized phase II (n = 1) or phase III (n = 17) trials. For metastatic breast cancer, the dose and schedule associated with the most favorable therapeutic index for paclitaxel was 175 mg/m2 given as a 3-hour infusion every 3 weeks, and docetaxel was 60-100 mg/m2 given as a 1-hour infusion every 3 weeks. Survival was improved under the following circumstances: (1) when 4 cycles of paclitaxel (175 mg/m2 every 3 weeks) was given following 4 cycles of conventional doxorubicin-cyclophosphamide for axillary node-positive operable breast cancer, (2) when trastuzumab was added to paclitaxel as first-line therapy for metastatic breast cancer that overexpressed HER2/neu, and (3) when docetaxel was given as second-line therapy for anthracycline-resistant disease. Although a survival benefit was found for taxanes as a component of first-line therapy in two of six trials, the interpretation of both positive trials was confounded by a lack of crossover to taxane therapy in those who were initially randomized to receive standard therapy. The taxanes improve survival in patients with early-stage breast cancer and selected patients with metastatic breast cancer. Further research is necessary in order to identify the efficacy of docetaxel relative to paclitaxel, the optimal dose of docetaxel, the role of weekly taxane therapy, the role of trastuzumab plus taxanes in early-stage disease, and whether taxanes are more effective when given concomitantly or sequentially in patients with early-stage disease. PMID- 11899389 TI - Treatment of metastatic bone disease in breast cancer: bisphosphonates. AB - Like other metastases, bone metastases in breast cancer patients are not only a sign of the incurable nature of the underlying disease, but are also associated with specific complications. In particular, bone pain and pathological fractures impair the quality of life of those affected. Any treatment concept must, therefore, place the highest priority on preventing or reducing skeletal complications. There are two treatment options--local and systemic. Local therapy includes radiotherapy as well as surgical and orthopedic measures. The four pillars of systemic treatment are hormone therapy, chemotherapy, antiresorptive therapy with bisphosphonates, and treatment with centrally and/or peripherally acting analgesics. A precondition for successful treatment is close cooperation between medical/clinical oncologists, radiotherapists, surgeons/orthopedists, gynecologists, pain specialists, and endocrinologists (in the presence of a hypercalcemic syndrome). Patients with breast cancer associated solely with osseous metastasis may live for a number of years. It is, therefore, all the more important to start appropriate therapeutic measures early. Bisphosphonates play a particularly valuable role, since their main effect lies in the prevention of skeletal complications. Rather than replacing antineoplastic therapy, this class of substances supplements other treatments. Once started, bisphosphonate therapy should be given for the remainder of the patient's life, even in the event of osseous progression. PMID- 11899390 TI - Tamoxifen-induced hot flashes. AB - Hot flashes are the most prominent side effect of tamoxifen, the most frequently prescribed antitumor agent in the world. Little detailed information is available to predict who will develop hot flashes on tamoxifen, to describe the natural history of these hot flashes, and/or to predict who will request therapy for such a side effect. This current trial was developed to address these items. Women who were about to begin adjuvant tamoxifen for locally treated breast cancer were approached for this trial. Before initiating tamoxifen, patients completed a short questionnaire designed to inquire about potential prognostic factors. Upon starting tamoxifen, women were asked to complete a hot flash diary daily for 3 months, and then daily for 1 week of each of the subsequent 9 months. Fifty patients, aged 51-83 years, provided data for this report. Approximately half of the women reported that they did not have any substantial hot flashes while the other half reported hot flashes of variable intensity. On average, these hot flashes gradually increased over 3 months and then plateaued. Baseline factors that appeared to predict for subsequent hot flash problems included a prior history of moderate to severe hot flashes with menopause and a history of prior estrogen therapy use. Overall, 16% of the women reported the desire for therapy for their hot flashes. Thirty-seven and one-half percent of the women (6/16) had a history of both prior estrogen use and moderate to severe hot flashes with menopause as compared to none (0/14) of the women without either of these factors. The data from this study can be utilized to better identify and educate women as to their probability of developing hot flashes after starting tamoxifen, to describe the average time frame for these hot flashes, and to predict the likelihood of whether resultant hot flashes will be substantial enough to have a woman request therapy for them. PMID- 11899391 TI - Prevalence of BRCA1 and BRCA2 mutations in male breast cancer patients in Canada. AB - Men who inherit a mutation in the BRCA2 gene carry a 6% risk of developing breast cancer by the age of 70. The proportion of male breast cancers attributable to BRCA mutations has not yet been determined with accuracy. We studied a series of 14 male breast cancer patients, unselected for family history or ethnicity, who were treated at a single regional cancer center in Canada. Family histories were obtained, and the men were tested for germ-line mutations of BRCA1 and BRCA2. Seven of these patients had a significant family history of breast cancer (i.e., at least one first- or second-degree relative with breast cancer diagnosed before age 70). Two of the men carried BRCA2 mutations, but no BRCA1 mutations were found. Both mutation carriers reported a positive family history and a personal history of cancer that preceded their diagnosis of breast cancer. Our results support the recommendation that male breast cancer patients who have a significant family history of breast or ovarian cancer should be offered genetic counseling and testing. PMID- 11899392 TI - Magnetic resonance imaging of occult breast cancer. AB - Magnetic resonance imaging (MRI) findings in a case of clinically and mammographically occult breast cancer are presented. Breast MRI allows detection of the site of primary cancer with high sensitivity in patients with occult neoplasms and should be considered as a routine test in these patients. PMID- 11899393 TI - A pilot study of neoadjuvant paclitaxel and radiation with correlative molecular studies in stage II/III breast cancer. AB - A commonly used approach to the management of locally advanced breast cancer currently involves neoadjuvant chemotherapy, followed by surgery and radiation. Earlier neoadjuvant regimens had utilized doxorubicin, making concurrent treatment with radiation less desirable given dose-limiting normal tissue toxicities. With the development of paclitaxel, we can now reconsider the use of concurrent chemoradiation in the treatment of breast cancer. Although paclitaxel is a known radiation sensitizer, its precise mechanism of action is still unclear. One of its proposed mechanisms is that it binds tubulin and induces an M phase arrest. As cells in M-phase are very sensitive to radiation, it thereby increases radiation sensitivity. The ability to predict tumor response for individual patients would allow us to tailor subsequent therapy for the individual patient. This study is designed to evaluate if paclitaxel's effects on the cell cycle of an individual patient can predict the responsiveness of that patient's tumor to paclitaxel and radiation. Patients will be treated with 3 cycles of paclitaxel followed by concurrent paclitaxel and radiation prior to definitive surgery. PMID- 11899394 TI - Underutilization of breast-conserving therapy in a predominantly rural population: need for improved surgeon and public education. AB - Though breast-conserving therapy (BCT) was first recommended as the preferred treatment for women with early-stage breast cancer in 1990, little is known about the factors influencing or limiting the use of BCT in rural women. We retrospectively surveyed all surviving patients (227) referred to the Roger Maris Cancer Center over a 2-year period. Disease characteristics were verified by the tumor registry and random chart review. Responses were obtained from 171 patients (75%), a median of 26 months from diagnosis. The majority of patients were from rural areas; only 32% resided in towns with a population greater than or equal to 15,000. Thirty-five percent of those patients meeting published criteria had BCT. Patients who underwent BCT were younger (mean age 56.8 vs. 62.5, P = 0.01), more likely to have benign axillary lymph nodes (82% vs. 64%, P = 0.008), and more likely to be employed away from the home (66% vs. 44%, P = 0.01) than patients who underwent mastectomy (MRM). Distance from the nearest radiation facility did not affect treatment decisions (mean: 59.5 miles BCT vs. 52.6 miles MRM). Most patients (83%) ranked their surgeon as the most important source of information about treatment options. Perceived surgical recommendations were generally followed. Only three patients who felt their surgeon recommended MRM underwent BCT; eleven patients chose MRM though they believed their surgeon recommended BCT. The choice of local therapy is predominantly a surgeon-driven process; logistical barriers unique to a rural population had little impact. Unfortunately, many surgeons continue to apply much more stringent criteria when recommending BCT than those in published guidelines. PMID- 11899395 TI - Thalidomide: when everything old is new again. AB - Thalidomide is an anti-angiogenesis agent that currently is being evaluated in the treatment of various types of cancer. The teratogenic effects of thalidomide are well-known, and patients who are prescribed this drug often are fearful of its effects. Because of the potential for teratogenicity, patients must adhere to the System for Thalidomide Education and Prescribing Safety (STEPS). Side effects of thalidomide include sedation, dose-related peripheral neuropathy, constipation, thrombiotic events, and skin rash. Nurses have a major role in educating patients about this drug, its effects, and necessary precautions. PMID- 11899396 TI - Details on demand: consumers, cancer information, and the Internet. AB - An increasing number of patients and their families have Internet access and use it to make sense of their cancer experience. Search engines and subject guides take Internet users to cancer-related Web sites that contain information of varying quality and accuracy. Several organizations have attempted to create evaluation tools for cancer information on the Internet. They are, for the most part, untested, but they can serve as a practical guide for coaching patients and family members on the use of the Internet. Evaluating Internet information with guidelines, dealing with Internet-savvy patients and families, and the use of e mail in clinical practice are discussed. PMID- 11899397 TI - Stevens-Johnson syndrome. AB - Stevens-Johnson Syndrome (SJS), or erythema multiforme, is a severe, acute, adverse, cutaneous reaction to certain medications, such as phenytoin and topical nitrogen mustard. The risk of developing SJS is high when phenytoin and steroids are administered during cranial irradiation. SJS produces headache, malaise, sore throat, fever, and sloughing of the skin and mucous membranes. Prompt recognition of SJS and withdrawal of the offending medication is key to treating this disorder. Nurses play an important role in assessing patients and educating them about signs and symptoms of SJS. PMID- 11899398 TI - The dance of life. PMID- 11899399 TI - Fever of unknown origin in patients undergoing chemotherapy. AB - Drug fever refers to a febrile response to a drug, and its clinical picture often resembles an allergic reaction or infection. The fever most commonly occurs 7-10 days after drug administration, persists as long as the drug is continued, and disappears soon after stopping the drug (Tabor, 1986). The risks and benefits of continuing a drug that causes fever must be evaluated for every patient who experiences drug fevers. Quality-of-life issues arise for patients who experience them despite the concurrent use of steroids. Recognizing drug fever is of great clinical importance. If drug fever is not recognized, patients may be subjected to prolonged hospitalizations and unnecessary testing and medications (Johnson & Cunha, 1996). Oncology nurses play an important role in the early recognition of drug fever. PMID- 11899400 TI - Prodrugs. PMID- 11899402 TI - Temozolomide (Temodar). PMID- 11899401 TI - Capecitabine (Xeloda). PMID- 11899403 TI - Give a little ... get a lot. PMID- 11899405 TI - Ascites. PMID- 11899404 TI - Mottled feet and a clogged blood analysis machine. PMID- 11899406 TI - Readers bring attention to value of surgical intervention in treating colon cancer. PMID- 11899407 TI - Nursing considerations for managing topotecan-related hematologic side effects. AB - Topotecan (Hycamtin, SmithKline Beecham, Philadelphia, PA) was approved by the U.S. Food and Drug Administration in 1996 for use in relapsed ovarian cancer and in 1999 for platinum-sensitive small-cell lung cancer. Hematologic toxicity has been the predominant side effect associated with its use. Patients who have had extensive platinum-based therapy have exhibited increased degrees of thrombocytopenia and more severe neutropenia. These adverse events can be managed by identifying high-risk patients (i.e., those with more than six cycles of chemotherapy containing an alkylating agent or radiation to more than 25% of marrow-bearing bones, patients with a history of myelosuppression or renal impairment) and by recommending appropriate dose modifications based on the creatinine clearance measurement. By reducing the topotecan dose, myelosuppressive effects, as evidenced by neutropenia and thrombocytopenia, may be lessened or prevented without reducing the antitumor response. PMID- 11899408 TI - Health ethics in cross-cultural settings. PMID- 11899409 TI - Infected cardiac myxoma: an unusual phenomenon. AB - Infected cardiac myxoma is a rare phenomenon. Cardiac myxomas are primary tumors of the heart, usually benign and account for about 50% of the benign cardiac tumors. Patients usually present with a triad of constitutional, embolic, and obstructive symptoms. There are few reported cases of infected cardiac myxoma and there is no widely used definition. This is a case presentation describing a patient who was admitted to our institution, underwent surgical excision, and was subsequently discharged home. PMID- 11899410 TI - Papillary fibroelastoma: a rare cardiac tumor (a case presentation). AB - Cardiac papillary fibroelastoma is a rare, benign, primary tumor of the heart and accounts for seventy-eight percent of cardiac tumors. Patient are usually asymptomatic although there is a significant risk for embolization. Papillary fibroelastomas usually arise from the cardiac valves. This is a case presentation of a patient who was admitted to our institution and his course of treatment leading to excision of the cardiac papillary fibroelastoma. PMID- 11899411 TI - Tonsillectomy. A comparative study of dissection/snare vs suction-cautery. AB - In an optimal situation, a surgical procedure would be one that generates minimal post-operative pain, incurs little or no bleeding, and allows the patient to return to their normal daily activities in the shortest time period. A tonsillectomy is one of the most common operations performed in the world. Various surgical procedures for tonsillectomy are performed with a wide array of opinions to support the pros and cons of each technique. OBJECTIVES/GOALS: To determine if there is a significant difference between two methods of tonsillectomy. METHODS AND MATERIALS: A prospective single blinded randomized control study using (i) A dissection/snare technique, and (ii) A suction-cautery method. Measured outcomes such as blood loss, surgical time, post-op pain, post op hydration, pyrexia, and the length of time to resume normal daily activities will be assessed. RESULTS: In total, 50 patients were studied, 23 in the dissection/snare technique, and 27 in the suction cautery technique. Inclusion criteria was, the patient must be at least 2 years of age and not older than 16 years of age. Data was collected intra-operatively, at 2 and 4 hour post-op intervals, as well as a 2 week follow-up questionnaire completed by the parents. CONCLUSIONS: The suction cautery group had statistically significant differences in blood loss, surgical time and pain in the immediate post-operative period. PMID- 11899412 TI - What do we mean by targeted therapy? PMID- 11899413 TI - Platinum compounds in the treatment of advanced breast cancer. AB - Interest in platinum compounds for the treatment of breast cancer has been reawakened because of preclinical studies indicating synergy of platinum salts with the monoclonal antibody trastuzumab in human breast cancer cell lines that overexpress HER2/neu. Cisplatin, carboplatin, and iproplatin are not very active as single agents in patients with previously treated metastatic breast cancer (MBC). The activity of oxaliplatin has not been adequately tested in refractory MBC. On the other hand, cisplatin is very active as first-line chemotherapy, with response rates (RR) of 50%; carboplatin appears to be moderately active in patients without prior chemotherapy (RR around 30%). The clinical effectiveness of the other platinum compounds (iproplatin, oxaliplatin, and others) has not yet been fully tested as first-line chemotherapy. Platinum compounds have been extensively tested in combination with other antitumoral agents. Cisplatin combinations have been employed as neoadjuvant chemotherapy in women with locally advanced breast cancer. These combinations are very active, although the precise contribution of cisplatin to the overall activity is not known. Combinations with cisplatin have been investigated, essentially, as salvage therapy for patients with previously treated MBC. The combinations of cisplatin with older pharmacological agents (5-fluorouracil, etoposide) have moderate activity, while the combinations of cisplatin with the newer agents (vinorelbine, paclitaxel, docetaxel, gemcitabine) appear to be more active. The combinations of carboplatin with the classical agents (5-fluorouracil, etoposide) are poorly active in previously treated MBC; however, the combination of carboplatin with the taxanes (docetaxel, paclitaxel) is more active. Of greatest interest is the synergy between the platinum derivatives and the monoclonal antibody trastuzumab demonstrated in vitro in breast cancer cell lines overexpressing HER2/neu. Currently, several combinations of platinum compounds (either cisplatin or carboplatin) with docetaxel and trastuzumab are under clinical testing in patients with MBC who overexpress HER2/neu. The preliminary results are very promising, and these combinations will soon be tested in the adjuvant setting. Cisplatin, carboplatin, and perhaps, oxaliplatin appear to have some antitumor activity in MBC and can be combined safely with other agents that are active in this disease. However, the precise role that platinum compounds play in the treatment of breast cancer remains to be defined. PMID- 11899414 TI - Tubular breast cancer experience at Washington University: a review of the literature. AB - Our objective was to review the presentation and management of patients with tubular breast cancer treated at Barnes-Jewish Hospital and compare our findings with the available literature. Of 3908 cases of breast cancer treated at our institution between 1986 and 1995, the incidence of tubular breast cancer at Barnes-Jewish Hospital was 1.25%. We reviewed the breast cancer risks, initial presentation, treatment, and outcome of 39 women with 40 tubular breast cancers and compared our series with others in the literature. The mean patient age was 67 years, which is older than most other series. Twenty-nine of the 39 cancers (74%) were detected by screening mammography; the remainder presented with a palpable breast mass. The mean tumor size was 8 mm (range, 1-60 mm). Twenty-three of 25 tumors were ER+ (92%) and none had axillary nodal involvement. Bilateral breast cancer developed in 3 patients (8%). An additional 500 cases of tubular breast cancer have been described in the literature. When the component of the invasive tumor is > 75% tubular carcinoma, most patients present with early-stage disease that is ER+ in 47 of 56 tumors (84%). The natural history is indolent and metastases are rare. Bilateral breast cancer developed in 58 of the 540 cases (11%), 4 of which were tubular carcinomas. Local recurrences developed in 9 of 29 patients (31%) treated by excision alone. The role of tamoxifen has not been determined. Given the available data, the initial surgical staging and management of tubular carcinoma should be identical to other invasive histologies. PMID- 11899415 TI - Breast cancer and human immunodeficiency virus: a report of 20 cases. AB - Carcinoma of the breast is the most common malignancy in women in the United States. More than 40% of patients with human immunodeficiency virus (HIV) infection develop cancer during their illness, but breast cancer has seldom been reported. Twenty patients with breast cancer and HIV infection seen at the University of Miami/Jackson Memorial Hospital between January 1988 and August 2000 were retrospectively analyzed. Seventeen patients had a previous or concurrent diagnosis of HIV at the time of the breast cancer diagnosis. Their CD4 count ranged from 13-1126/microL (median, 309/microL). Most patients were premenopausal (16 of 20), with ages ranging from 31-61 years (median, 44 years). All stages of breast cancer were seen: ductal carcinoma in situ (2 patients), stage I (1 patient), stage II (9 patients), stage III (6 patients), and stage IV (2 patients). Ten tumors had estrogen receptors. Four of the 13 patients who underwent axillary lymph node dissection had abnormal lymph node findings, including 2 with follicular hyperplasia and 2 with caseating granulomas. Seven patients received chemotherapy with very poor tolerance. Estrogen receptor positive patients were treated with tamoxifen. Of the 18 patients who presented with local disease, 7 have died: 2 of breast cancer, 4 of acquired immunodeficiency syndrome, and 1 of cardiac arrest. Nine patients remain free of disease (5 of them > 5 years) and 2 patients are alive with metastatic disease. Breast cancer in the HIV-positive population is similar to that seen in seronegative women. Most of the patients that are long-term survivors were treated with surgery and tamoxifen. The benefits of adjuvant chemotherapy are not clear. PMID- 11899416 TI - Molecular determinants of occult metastatic tumor cells in bone marrow. AB - The success of mammographic screening for breast cancer is that it involves increasingly more patients with small primary tumors formerly thought to have an overall excellent prognosis. Yet, only approximately two thirds of these patients actually have this favorable prognosis, while the remaining third develops metastatic disease. Thus, there is emerging evidence that epithelial tumor cells can disseminate into secondary organs at an earlier stage of primary tumor development than appreciated by current risk classifications. Bone marrow is one of the most prominent secondary organs screened for the presence of disseminated tumor cells. The current data suggest that bone marrow micrometastases represent a selected population of dormant and heterogeneous cancer cells. The analysis of micrometastatic cells opens a new avenue by which to assess the molecular determinants of both early tumor cell dissemination and subsequent outgrowth into overt metastases. Moreover, identifying therapeutic target structures (e.g., HER2/neu), monitoring the elimination of bone marrow micrometastases, and assessing treatment-resistant tumor cell clones might help to understand the current limitations of adjuvant systemic therapy. This review summarizes the current knowledge of the biological characteristics of micrometastatic cancer cells in bone marrow of breast cancer patients. PMID- 11899417 TI - Detection of occult disease in breast cancer using fluorodeoxyglucose camera based positron emission tomography. AB - An isolated increase of blood tumor marker CA 15.3 in breast cancer is considered a sensitive indicator for occult metastatic disease but by itself is not sufficient for initiating therapeutic intervention. We investigated the potential of camera-based positron emission tomography (PET) imaging using [18F] fluorodeoxyglucose (FDG) to detect clinically occult recurrences in 132 female patients (age, 35-69 years) treated for breast cancer, all presenting with an isolated increase in blood tumor marker CA 15.3 without any other evidence of metastatic disease. FDG results were correlated to pathology results or to a sequentially guided conventional imaging method. One hundred nineteen patients were eligible for correlations. Positive FDG scans were obtained for 106 patients, including 89 with a single lesion and 17 with 2 or more lesion. There were 92 true-positive and 14 false-positive cases, 10 of which became true positive within 1 year. Among the 13 negative cases, 7 were false negative and 6 were true negative. Camera-based PET using FDG has successfully identified clinically occult disease with an overall sensitivity of 93.6% and a positive predictive value of 96.2%. The smallest detected size was 6 mm for a lymph node metastasis (tumor to nontumor ratio, 4:2). FDG camera-based PET localized tumors in 85.7% of cases suspected for clinically occult metastatic disease on the basis of a significant increase in blood tumor marker. A positive FDG scan associated with an elevated CA 15.3 level is most consistent with metastatic relapse of breast cancer. PMID- 11899418 TI - Use of intrathecal trastuzumab in a patient with carcinomatous meningitis. PMID- 11899420 TI - Antigen-specific immunotherapy for autoimmune disease: fighting fire with fire? PMID- 11899419 TI - Commentary on "A review of vinorelbine in the treatment of breast cancer". PMID- 11899422 TI - Inhibition of human T-cell responses by allergen peptides. PMID- 11899421 TI - Peptide-based immunotherapy of autoimmunity: a path of puzzles, paradoxes and possibilities. PMID- 11899423 TI - Beneficial effect of co-polymer 1 on cytokine production by CD4 T cells in multiple sclerosis. AB - Multiple sclerosis (MS) has been associated with an imbalance in the T helper type 1 (Th1) and Th2 subsets. We investigated, at the single-cell level, the synthesis of pro-inflammatory cytokines by CD4 and CD8 T cells from MS patients. We report the relationship between priming of CD4 and CD8 T cells for interleukin 2 (IL-2), interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF alpha) and disease evolution in MS patients, clinically subdivided into relapsing remitting MS (RRMS) in remission, RRMS in relapse, or chronic progressive MS (CPMS). Moreover, we report the in vivo influence of co-polymer 1 (COP) treatment on the pattern of cytokine producers in RRMS patients. We show that the frequency of CD4 T cells primed for TNF-alpha synthesis increased in all stages of MS, including RRMS remitting, and was normalized to control values in COP-treated patients (43.2 +/- 11.8% in treated patients versus 47 +/- 7.3% in RRMS remitting versus 40.3 +/- 8% in controls). In addition, a significant decrease in the frequency of CD4 T cells primed for IL-2 was found in COP-treated patients as compared to the other groups of patients, reaching values below that of controls (59.1 +/- 9.9% in treated patients versus 70 +/- 11.6% in RRMS remitting versus 67.1 +/- 7.4% in controls). Unexpectedly, COP-treated patients also showed a significantly decreased priming for IFN-gamma at the CD4 T-cell level (9.1 +/- 3.4% in treated patients versus 18.8 +/- 0.6.4% in RRMS remitting versus 15.4 +/- 4.7% in controls), but not at the CD8 T-cell level. This bystander suppression on the inflammatory cells should be considered in the monitoring of MS patients submitted to COP treatment, in order to evaluate better its clinical efficacy. PMID- 11899424 TI - Requirements for autoimmune responses to mouse gastric autoantigens. AB - Autoimmune gastritis, in which the H+/K(+)-ATPase of parietal cells is the major antigen, is one of the most common autoimmune diseases. Here we examined if specific properties of the H+/K(+)-ATPase or parietal cells are involved in rendering them autoimmune targets. The model antigens beta-galactosidase and ovalbumin (OVA) were expressed in parietal cells of transgenic mice. On experimental induction of autoimmune gastritis by neonatal thymectomy, autoantibodies to beta-galactosidase developed in mice expressing beta galactosidase in parietal cells, a response that was independent of either the response to the gastric H+/K(+)-ATPase or gastric inflammation. In contrast, mice that expressed OVA in parietal cells did not exhibit an antibody response to OVA after thymectomy. However, increasing the frequency of anti-OVA T lymphocytes in OVA-expressing mice resulted in autoantibodies to OVA and gastritis. These studies indicate that parietal cells can present a variety of antigens to the immune system. Factors such as the identity and expression level of the autoantigen and the frequency of autoreactive T cells play a role in determining the prevalence and outcome of the particular immune response. In addition, as not all mice of a particular genotype displayed autoimmunity, random events are involved in determining the target of autoimmune recognition. PMID- 11899426 TI - B-cell precursors differentiated from cord blood CD34+ cells are more immature than those derived from granulocyte colony-stimulating factor-mobilized peripheral blood CD34+ cells. AB - Umbilical cord blood (CB) has been widely used instead of bone marrow (BM) and peripheral blood (PB) for stem cell transplantation (SCT). However, problems of sustained immunodeficiency after CB transplantation remain to be resolved. To elucidate the mechanism of immunodeficiency, we compared the characteristics of B cells differentiated in vitro from CD34+ cells of CB with those of PB. Purified CD34+ cells from CB and PB were cultured on murine stroma cell-line MS-5 with stem cell factor and granulocyte colony-stimulating factor for 6 weeks. The B cell precursors (pre-B cells) that differentiated in this culture system, were analysed as to their immunoglobulin heavy chain (IgH) variable region gene repertoire and the expression of B-cell differentiation-related genes. CD10+ CD19+ pre-B cells were differentiated from both PB and CB. Although the usages of IgH gene segments in pre-B cells differentiated from CB and PB were similar, the N region was significantly shorter in CB-derived than PB-derived cells. Productive rearrangements were significantly fewer in cells of CB than PB in the third week. Among a number of B-cell differentiation-related genes, the terminal deoxynucleotidyl transferase (TdT) gene was not expressed in CB-derived cells during the culture. These results indicated that immature features of pre-B cells from CB, such as lack of TdT expression, and a short N region and few productive rearrangements in the IgH gene, might cause the delay in mature B-cell production. PMID- 11899425 TI - Interleukin-1 beta, but not interleukin-1 alpha, is required for T-cell-dependent antibody production. AB - Interleukin-1 (IL-1) consists of two molecules, IL-1 alpha and IL-1 beta, and IL 1 receptor antagonist (IL-1Ra) is a natural inhibitor of these molecules. Although the adjuvant effects of exogenously administered IL-1 in the humoral immune response are well known, the roles of endogenous IL-1 and the functional discrimination between IL-1 alpha and IL-1 beta have not been elucidated completely. In this report, we investigated the role of IL-1 in the humoral immune response using gene-targeted mice. Both primary and secondary antibody production against T-dependent antigen, sheep red blood cells (SRBC), was significantly reduced in IL-1 alpha/beta-/- mice, and was enhanced in IL-1Ra-/- mice. The intrinsic functions of B cells, such as antibody production against type 1 T-independent antigen, trinitrophenyl-lipopolysaccharide and proliferative responses against mitogenic stimuli, were normal in IL-1 alpha/beta-/- mice. The proliferative response of T cells and cytokine production upon stimulation with anti-CD3 monoclonal antibody were also normal, as was the phagocytotic ability of antigen-presenting cells (APCs). However, SRBC-specific proliferative response and cytokine production of T cells through the interaction with APCs were markedly impaired in IL-1 alpha/beta-/- mice, and enhanced in IL-1Ra-/- mice. Moreover, we show that SRBC-specific antibody production was reduced in IL-1 beta /- mice, but not in IL-1 alpha-/- mice. These results show that endogenous IL-1 beta, but not IL-1 alpha, is involved in T-cell-dependent antibody production, and IL-1 promotes the antigen-specific T-cell helper function through the T-cell APC interaction. PMID- 11899427 TI - CR2-mediated activation of the complement alternative pathway results in formation of membrane attack complexes on human B lymphocytes. AB - Normal human B lymphocytes activate the alternative pathway of complement via complement receptor type 2 (CR2, CD21), that binds hydrolysed C3 (iC3) and thereby promotes the formation of a membrane-bound C3 convertase. We have investigated whether this might lead to the generation of a C5 convertase and consequent formation of membrane attack complexes (MAC). Deposition of C3 fragments and MAC was assessed on human peripheral B lymphocytes in the presence of 30% autologous serum containing 4.4 mM MgCl2/20 mM EGTA, which abrogates the classical pathway of complement without affecting the alternative pathway. Blockade of the CR2 ligand-binding site with the monoclonal antibody FE8 resulted in 56 +/- 13% and 71 +/- 9% inhibition of the C3-fragment and MAC deposition, respectively, whereas the monoclonal antibody HB135, directed against an irrelevant CR2 epitope, had no effect. Blockade of the CR1 binding site with the monoclonal antibody 3D9 also resulted in a minor reduction in MAC deposition, while FE8 and 3D9, in combination, markedly reduced deposition of both C3 fragments (91 +/- 5%) and C9 (95 +/- 3%). The kinetics of C3-fragment and MAC deposition, as well as the dependence of both processes on CR2, indicate that MAC formation is a consequence of alternative pathway activation. PMID- 11899428 TI - Blockage of complement regulators in the conjunctiva and within the eye leads to massive inflammation and iritis. AB - The open environment of the eye is continuously subject to an influx of foreign agents that can activate complement. Decay-accelerating factor (DAF), membrane cofactor protein (MCP) and CD59 are regulators that protect self-cells from autologous complement activation on their surfaces. They are expressed in the eye at unusually high levels but their physiological importance in this site is unstudied. In the rat, a structural analogue termed 5I2 antigen (5I2 Ag) has actions overlapping DAF and MCP. In this investigation, we injected F(ab')2 fragments of 5I2 mAb into the conjunctiva and aqueous humor, in the latter case with and without concomitant blockage of CD59. Massive neutrophilic infiltration of the stroma and iris resulted upon blocking 5I2 Ag activity. Frank necrosis of the iris occurred upon concomitant intraocular blockage of CD59. C3b was identified immunohistochemically, and minimal effects were seen in complement depleted animals and in those treated with non-relevant antibody. The finding that blockage of 5I2 Ag function in periocular tissues and within the eye causes intense conjunctival inflammation and iritis demonstrates the importance of intrinsic complement regulators in protecting ocular tissues from spontaneous or bystander attack by autologous complement. PMID- 11899429 TI - Redox imbalance and immune functions: opposite effects of oxidized low-density lipoproteins and N-acetylcysteine. AB - This study investigates the in vitro effects of oxidized low-density lipoproteins (ox-LDL), 'physiological' pro-oxidants, N-acetylcysteine (NAC), a free radical scavenger and glutathione precursor, and their combination on human peripheral blood mononuclear cell functions. We found that treatment with ox-LDL induced a significant down-regulation of proliferative response to mitogens, antigens and interleukin-2. Lipid extracts from ox-LDL were able to reproduce the same effect as the lipoprotein. On the other hand, NAC exposure induced a significant up regulation of proliferative responses to all the stimuli used. Moreover, we showed that natural killer (NK) cell-mediated cytotoxic activity was significantly down-regulated by ox-LDL while treatment with NAC induced a significant up-regulation of NK-cell activity. Finally, we found that ox-LDL and NAC exerted opposite effects on the cytokine network, interfering both at the protein secretion level and the messenger RNA synthesis level. More importantly, when NAC was used in combination with ox-LDL the proliferative responses, NK-cell mediated cytotoxic activity and cytokine production were restored to values comparable to controls. These data indicate that ox-LDL and NAC modulate immune functions, exerting opposite effects reflecting their pro-oxidant and antioxidant behaviours. Our results add new insights to the key role played by redox imbalance as a modulator of immune system homeostasis and suggest that an antioxidant drug such as NAC could be useful against pathologies associated with an increase in lipid peroxidation. PMID- 11899430 TI - Adrenaline suppression of the macrophage nitric oxide response to lipopolysaccharide is associated with differential regulation of tumour necrosis factor-alpha and interleukin-10. AB - Adrenaline is a catecholamine hormone secreted by the adrenal medulla in response to acute stress. Previous studies have shown that adrenaline suppresses the nitric oxide (NO) response of murine macrophages (M phi s) stimulated in vitro with lipopolysaccharide (LPS). We have now extended these studies to examine the effects of adrenaline on the production of tumour necrosis factor alpha (TNF alpha) and interleukin-10 (IL-10). Our results showed that NO, TNF-alpha and IL 10 were concurrently produced following in vitro LPS (10 micrograms/ml) stimulation of murine peritoneal M phi s. Adrenaline suppressed both NO and TNF alpha with concomitant up-regulation of the IL-10 response above that seen with LPS alone. In this in vitro model of LPS stimulation we demonstrated that TNF alpha was required for NO production, as the TNF-alpha neutralizing monoclonal antibody, TN3.19.12, abolished the response; in contrast, IL-10 suppressed NO. In order to determine any functional consequence of adrenaline-mediated IL-10 augmentation on NO production, M phi s were stimulated with LPS and specific neutralizing anti-IL-10 antibodies were added to the cultures. The LPS NO response was suppressed to 43% of the control value by adrenaline (10(-8) M) and an irrelevant control antibody had no effect on the adrenaline-mediated inhibition of NO, but anti-IL-10 treatment restored the NO response to levels similar to those observed with LPS alone. Furthermore, we demonstrated that exogenous TNF-alpha, at a dose range of 1.9-50 ng per ml, also restored the nitrite response to LPS in the presence of adrenaline. Together, the observations that neutralization of IL-10 and addition of TNF-alpha abrogate adrenaline's inhibition of NO, suggest that this hormone suppresses NO partly through up regulation of IL-10 which, in turn, may suppress TNF-alpha that is required for NO production. Finally, we also observed that the M phi-activating cytokine, interferon-gamma (IFN-gamma), attenuated the inhibitory effect of adrenaline on the LPS NO response. PMID- 11899431 TI - Nitric oxide-enhanced resistance to oral candidiasis. AB - A murine model of oral candidiasis was used to show that nitric oxide (NO) is involved in host resistance to infection with Candida albicans in infection 'resistant' BALB/c and infection-'prone' DBA/2 mice. Following infection, increased NO production was detected in saliva. Postinfection samples of saliva inhibited the growth of yeast in vitro. Treatment with NG-monomethyl-L-arginine (MMLA), an inhibitor of NO synthesis, led to reduced NO production, which correlated with an increase in C. albicans growth. Reduction in NO production following MMLA treatment correlated with an abrogation of interleukin-4 (IL-4), but not interferon-gamma (IFN-gamma), mRNA gene expression in regional lymph node cells. Down-regulation of IL-4 production was accompanied with an increase in IFN gamma production in infection-'prone' DBA/2 mice. There was a functional relationship between IL-4 and NO production in that mice treated with anti-IL-4 monoclonal antibody showed a marked inhibition of NO production in saliva and in culture of cervical lymph node cells stimulated with C. albicans antigen. The results support previous conclusions that IL-4 is associated with resistance to oral candidiasis and suggest that NO is involved in controlling colonization of the oral mucosal surface with C. albicans. PMID- 11899432 TI - Human immunodeficiency virus type 1 Tat binds to Candida albicans, inducing hyphae but augmenting phagocytosis in vitro. AB - Tat, the human immunodeficiency virus type 1 (HIV-1) transactivating protein, binds through its RGD-motif to human integrin receptors. Candida albicans, the commonest cause of mucosal candidiasis in subjects infected with HIV-1, also possesses RGD-binding capacity. The present study reveals that Tat binds to C. albicans but not to C. tropicalis. Tat binding was markedly reduced by laminin and to a lesser extent by a complement C3 peptide containing the RGD motif, but not by a control peptide. The outgrowth of C. albicans was accelerated following binding of Tat, but phagocytosis of opsonized C. albicans was also increased after Tat binding. Thus, Tat binding promotes fungal virulence by inducing hyphae but may also reduce it by augmenting phagocytosis. The net effect of Tat in vivo is difficult to judge but in view of the many disease-promoting effects of Tat we propose that accelerating the formation of hyphae dominates over the augmentation of phagocytosis. PMID- 11899433 TI - Leishmania major lipophosphoglycan modulates the phenotype and inhibits migration of murine Langerhans cells. AB - Langerhans cells (LC), members of the dendritic cell family, play a central role in the initiation and regulation of the immune response against the protozoan parasite Leishmania major. LC take up antigens in the skin and transport them to the regional lymph nodes for presentation to T cells. However, it is not known whether LC functions are modulated by parasite antigens. In the present study, we examined the effect of a major parasite surface molecule, L. major lipophosphoglycan (LPG), on the maturation of LC and their migratory properties. The results show that exposure to LPG did not affect the expression of major histocompatibility complex (MHC) class II and B7, but induced an up-regulation of CD25, CD31 and vascular endothelial (VE)-cadherin expression and a down regulation of Mac-1 expression, by LC. Importantly, LPG treatment inhibited the migratory activity of LC, as it reduced their efflux from skin explants and their migration in transwell cultures. These results suggest that Leishmania LPG impairs LC migration out of the skin and thus may modulate their immunostimulatory functions, which require LC translocation from skin to lymph nodes. PMID- 11899435 TI - Delivering OR staff beyond the basic orientation. PMID- 11899436 TI - Liposuction. An update on one of the most frequently performed and controversial surgeries. PMID- 11899434 TI - Infection of murine keratinocytes with herpes simplex virus type 1 induces the expression of interleukin-10, but not interleukin-1 alpha or tumour necrosis factor-alpha. AB - Herpes simplex virus (HSV) is known to possess several mechanisms whereby it can evade the normal host immune defences. In this study the expression of the immunosuppressive cytokine, interleukin (IL)-10, was monitored following infection of a murine keratinocyte cell line (PAM-212) and compared with the expression of two proinflammatory cytokines: IL-1 alpha and tumour necrosis factor (TNF)-alpha. The PAM-212 cells were infected at a multiplicity of 0.5 with a clinical isolate of HSV type 1, and the mRNA of the three cytokines was assessed by semiquantitative reverse transcription-polymerase chain reaction (RT PCR) over the following 24 hr. By 12 hr postinfection the amount of IL-10 mRNA had increased significantly to five-fold greater than that found in uninfected cells (P < 0.01), and this elevated level was maintained until at least 24 hr postinfection. In contrast, IL-1 alpha and TNF-alpha mRNAs were not significantly up-regulated by the HSV infection. Immunostaining with an IL-10 monoclonal antibody (mAb) revealed that cytoplasmic IL-10 protein had increased by 6-12 hr postinfection. This quantity was further increased at 24 hr postinfection, when the viral cytopathic effect was apparent. Viral replication was necessary, but not sufficient on its own, for IL-10 induction. Experiments with HSV mutants lacking functional transactivating factors suggested that the viral transactivating proteins ICP-0 and VP-16 may be necessary for HSV-induced IL-10 expression. Thus, the up-regulation in the expression of IL-10 mRNA and protein induced by HSV early in the infection of keratinocytes represents a specific response and may be part of the viral strategy to avoid local immune defence mechanisms in the skin. PMID- 11899437 TI - Harassment in the workplace. PMID- 11899438 TI - Ethical competence and perioperative nursing. PMID- 11899439 TI - Nursing labour force--a national issue. PMID- 11899440 TI - Interim position statement on intensive care nursing staffing. PMID- 11899441 TI - ACCCN national nursing workforce survey of intensive care units. AB - A descriptive study was designed and implemented by the Australian College of Critical Care Nurses (ACCCN) Workforce Planning Advisory Committee to capture data pertaining to workforce issues of intensive care nurses. All intensive care units (ICUs) within Australia were mailed a self reporting survey. Despite a low response rate (52 per cent) and difficulty reported by respondents in gaining the appropriate data requested, the results revealed an interesting snapshot of the intensive care nursing workforce. Types of services offered by units varied considerably; paid overtime hours were low (< 2 per cent of total hours worked) and use of both part-time and agency staff was also low (10 per cent of total hours worked). Private hospitals utilised a greater proportion of part-time and agency nursing staff than public hospitals (20:10 per cent). The turnover rate for registered nursing staff was estimated at 18 per cent, with education, skill acquisition and improved communication reported as the major incentives used by managers to attract and retain staff. This study demonstrated that valuable data are currently uncaptured and recommends a more refined process of a national database to record and manage this important information for future workforce planning. PMID- 11899442 TI - Critical care dependency tool: monitoring the changes. AB - The critical care patient dependency system (CCPD) is a factorial patient acuity system developed in 1993 by Ferguson and Harris-Ingall' for use in adult critical care areas. It was developed specifically to help determine Australian nursing cost weights and was utilised to collect data from nine Sydney critical care units from October 1992 until May 1993. The St. George Hospital (SGH) general intensive care unit, one of the nine participating hospitals, continues to use and collect data with the CCPD. This paper describes the instrument and compares data on Australian national diagnosis related groups (ANDRGs), collected during the original study, to ANDRG information on the critical care population 3 and 6 years later. In addition, the paper examines and compares the demographics of the SGH critical care patient population, patient acuity (based upon CCPD patient scores) and intensive care nursing clinical practices collected over a 3 month period in 1996 and again in 1999. Demographic and patient acuity data for SGH in 1993 are unavailable and so comparisons were unable to be made. The findings demonstrate changes in the management of critically ill patients, especially in relation to ventilation management, wound care and invasive monitoring practices; this resulted in shifts to the nursing workload. For this reason, the instrument is useful in providing nurse managers with information about patient dependencies and nursing work. PMID- 11899443 TI - Preparing for coronary angioplasty: the patients' experiences. AB - Coronary angioplasty and stent placement procedures now represent one of the fastest growing specialties in cardiac care; patients undergo a short stay admission with limited care time with nurses. The purpose of this study was to describe participants' experiences of preparing for angioplasty in such an environment. Eight men and three women were interviewed 1 month after discharge from hospital. Verbatim transcripts were analysed for major themes using the qualitative techniques of grounded theory. Participants described working through a problem solving process in response to the perceived health threat associated with undergoing angioplasty. In step one, the problem was identified. In step two, coping responses were taken to try and solve the problem. In step three, the results of the coping responses were appraised or evaluated. The two problems identified were ongoing chest pain and anxiety related to fear of the unknown. The coping responses initiated included acquiring knowledge of the angioplasty, confidence in the skill of the doctor, support from family and gearing up psychologically. In the final appraisal of the coping responses, the participants decided to either go ahead with, or delay the angioplasty procedure. The results of this study indicate that the preparation for angioplasty represents a period of adjustment that may be anxiety provoking. Participants' experiences provide new knowledge of the concerns and challenges faced when undergoing such an invasive procedure in a short stay environment. The results clearly highlight that psychosocial aspects of nursing care are an essential component of nursing practice for angioplasty patients. PMID- 11899444 TI - Disseminated intravascular coagulation (DIC). AB - Disseminated intravascular coagulation (DIC) is a condition that is seen in critical care areas in association with a number of precipitating conditions. DIC is often described as a paradoxical disease process of concurrent haemorrhage and thrombus formation, resulting in a range of clinical manifestations related to end organ ischaemia. This article will discuss the coagulation cascade as a basis for understanding the aetiology and pathophysiology of DIC. The discourse surrounding medical treatment will be highlighted to enable bedside nurses to understand the divergent orders they may receive from physicians for different patients. Management of the patient experiencing DIC will be approached from a multidisciplinary focus, as well as highlighting the specific nursing considerations and alternatives from an independent focus. PMID- 11899445 TI - Examining critical care nurses' critical incident stress after in hospital cardiopulmonary resuscitation (CPR). AB - The object of this study was to determine if critical care nurses' emotional responses to having performed cardiopulmonary resuscitation were indicative of critical incident stress. A descriptive approach was employed using a survey questionnaire of 31 critical care nurses, with supportive interview data from 18 of those participants. Analysis of the data generated from the questionnaire indicated that the respondents experienced thought intrusion and avoidance behaviour. A majority of those interviewed disclosed that they had experienced a wide range of emotional stressors and physical manifestations in response to having performed the procedure. The findings from both questionnaire and interview data were congruent with signs of critical incident stress, as described in the literature. This has been found to be detrimental to employees' mental health status and, for this reason, employers have a duty of care to minimise the risk of its occurrence and to manage problems as they arise. PMID- 11899446 TI - Safety concerns, working with students and new graduates. Top trends in calls to AARN practice consultants. PMID- 11899448 TI - Reflective practice. PMID- 11899447 TI - AARN 2001 award winners. Recognizing outstanding registered nursing practice. PMID- 11899449 TI - Alberta RN demographics. AB - In 2000 more than one-quarter of the Alberta RN labour force was older than 50 years of age, while close to one-half was between 36 and 50 years of age. While this age distribution denotes a mature RN cadre, with considerable valuable practice experience, it also signals a need for an increased number of new RN graduates to replace those soon to be retiring. Although possibly age-related, the proportion of Alberta RNs working regular full-time is less than half--only 45 per cent. Concomitantly, more than half of all Alberta RNs are employed casually and part-time. Especially in a time of RN shortage, reasons for such limited full-time employment warrant examination. As in the past, three-quarters of Alberta RNs work in institutional settings--hospitals and nursing homes--while only eight per cent work in community health. The shift to health promotion and disease prevention has yet to happen in Alberta. In 2000, almost 80 per cent of all Alberta RNs were employed in direct care giving, and less than 10 per cent were employed in management, administration and education. In 2000, almost two thirds of all Alberta RNs held a diploma in nursing. Of the slightly more than 8,000 Alberta RNs with a baccalaureate degree, the overwhelming majority--85 per cent--worked in direct patient care, and only 13 per cent were employed in administration and teaching. Increasingly, we can expect to see more RNs with baccalaureate degrees at the bedside, in part because of the recent significant shift toward new graduates entering practice with a degree. Since 1997, the number of new RNs entering practice with a degree has been four times greater than the number entering with a diploma. Increased government funding of degree nursing programs aimed at alleviating the current nursing shortage means this trend will continue. PMID- 11899450 TI - Alberta RN on-line. PMID- 11899451 TI - Welcoming new graduate RNs. PMID- 11899452 TI - Improved method of applying a safety strap to prevent ocular injury from a Mayfield horseshoe apparatus. PMID- 11899453 TI - Assessing pain responses during general anesthesia. PMID- 11899454 TI - Assessing pain responses during general anesthesia. PMID- 11899455 TI - Anesthesia personnel and the Americans with Disabilities Act. PMID- 11899456 TI - TheNational Commission on Nurse Anesthesia Education 10 years later--Part 1: The commission years (1989-1994). AB - The significant decrease in the number of anesthesia providers during the late 1980s prompted American Association of Nurse Anesthetists (AANA) leaders to establish the National Commission on Nurse Anesthesia Education (NCNAE). The NCNAE was charged with scrutinizing all aspects of nurse anesthesia educational programs and developing strategies to reverse the critical shortage of nurse anesthetists. The tactics outlined by the commissioners were implemented, and they resulted in an increase of annual nurse anesthesia program graduates. Although there has been continued realization of NCNAE strategies, 10 years later the critical shortage of CRNAs has resurfaced. This 2-part article describes the commission years, the years that followed, and the current status of Certified Registered Nurse Anesthetist (CRNA) manpower. PMID- 11899458 TI - Anesthesia in a child with Rett syndrome: a case report and literature review. AB - Rett syndrome is an increasingly diagnosed syndrome in young children who appear normal at birth and develop normally until 6 to 18 months of age, when developmental milestones fail to be reached. The syndrome appears only in girls and therefore it is thought to be an X-linked dominant trait that is lethal in the male. This is a case report and literature review of anesthesia in a child with Rett syndrome. PMID- 11899457 TI - Preventable adverse patient outcomes: a closed claims analysis of respiratory incidents. AB - Closed claims analysis of adverse anesthesia outcomes was initiated through the AANA Foundation in 1995 to examine adverse outcomes of anesthesia care provided by Certified Registered Nurse Anesthetists (CRNAs). A research team of 8 CRNAs using an instrument incorporating more than 150 variables undertook document analyses of closed claim files. All files reviewed involved incidents in which the CRNA named in the policy was potentially involved in the adverse patient outcome. Thirty-eight percent (58/151) of CRNA-related claims involved a respiratory incident as the primary cause of the negative patient outcome. Patient outcomes involving respiratory incidents were more likely to result in death or permanent injury compared with nonrespiratory incidents (P < .01). Reviewers found that respiratory claims were more likely to have involved inappropriate anesthesia management (P < .01), more likely to have involved a lack of vigilance (P < .01), and more likely to have been judged by the reviewer as preventable (P < .01). A higher percentage of respiratory incidents occurred in emergency cases (75% vs 34%, P < .01) and in cases involving general anesthesia (44% vs 17%, P < .01). Adverse respiratory incidents are largely preventable and frequently result in serious patient morbidity and mortality. PMID- 11899459 TI - Transient radiculopathy after 5% lidocaine or 0.75% bupivacaine spinal anesthesia in 3 surgical positions. AB - The present study was conducted to compare the incidence of transient radicular irritation (TRI) after spinal anesthesia with 5% lidocaine or 0.75% bupivacaine in the supine, prone, and lithotomy surgical positions. A non-rAndomized survey approach was used. The convenience sample consisted of 243 adults receiving spinal anesthesia for elective surgery at 1 of 3 hospitals. Patients were questioned by telephone postoperatively to determine whether they had experienced TRI. Statistical analysis using the Fisher exact test revealed no significant difference in TRI incidence between local anesthetics in the supine or prone position groups. In the lithotomy position group, the incidence of TRI was significantly higher in patients receiving 5% lidocaine. Further, chi 2 testing revealed no significant difference in TRI incidence between surgical position groups when position alone was considered. The findings suggest that TRI after spinal anesthesia occurs more frequently with 5% lidocaine than with 0.75% bupivacaine only when patients undergo surgery in the lithotomy position. Providers need to consider the risks and benefits of 5% lidocaine when selecting an agent for spinal anesthesia, especially with patients undergoing surgery in the lithotomy position. When lidocaine is used, providers should discuss TRI as a risk of spinal anesthesia with patients during preanesthetic counseling. PMID- 11899460 TI - Analysis of noteworthy indicators on the anesthesia record: a prospective, multiregional study. AB - A national sample of anesthesia records were studied prospectively for the presence or absence of predetermined, noteworthy record indicators judged important in documenting a basic core of information describing the monitoring and care provided in a generic clinical setting. Only records involving the administration of a volatile anesthetic for the purpose of achieving general anesthesia, prepared by providers with at least 6 months of experience in anesthesia care were examined. No identifying patient, provider, or institutional data were recorded. Indicators were scored in a "present/absent" format. All data were pooled; the goal was to describe the overall phenomena in terms of frequency of compliance. A total of 4,989 anesthesia records were evaluated in terms of 13 record indicators. Those missed ranged from 0.0% (patient identification) to 28.6% (notation regarding emergence). Ten were omitted on less than 6% of the records: 3 (surgery and/or anesthesia start time, notation regarding emergence, surgical positioning noted) were omitted on more han 13.0% of the records. Given he observed inconsistencies, perhaps the anesthesia record needs redesign or the rationale behind documentation requires increased emphasis in educational and equally assurance programs. PMID- 11899462 TI - Not just for healthy bones and teeth. PMID- 11899461 TI - Latex allergy: are you at risk? AB - The prevalence of latex allergies in healthcare workers has significantly increased over the past 2 decades. Increases in exposure to latex products in healthcare environments are related primarily to the use of gloves for barrier protection. In the early 1980s, with the implementation of universal precautions and appropriate healthcare worker protection, latex glove use dramatically rose in many countries. Manufacturing techniques and additional factories for latex gloves were developed to meet the tremendous demand. As a result of new "fast track" production processes, some of the latex gloves had variant amounts of powder and allergen content. Synthetic materials, such as polyvinyl, polyurethanes, nitrile, and neoprene, are being used to manufacture medical gloves. Some hospitals and clinics have adopted the use of these alternatives to provide a "latex-free" healthcare environment. Risk reduction and prevention strategies are being implemented in many countries. Latex sensitivity and allergy can present as variable clinical reactions including contact dermatitis, rhinoconjuctivitis, asthma, and anaphylaxis. Some healthcare providers who have coexisting risk factors, such as atopy and food allergies (chestnuts, bananas, avocados, passion fruit, celery, potatoes, and peaches), are at an even greater risk for severe allergic reactions following repeated latex exposure. This journal course will provide an overview of the information available related to latex allergy in healthcare workers. PMID- 11899463 TI - Shiftwork. How to cope with life in the shadows. PMID- 11899464 TI - A midwife to the dying. PMID- 11899465 TI - Nontraditional microbial bioactive metabolites. AB - Microorganisms produce low-molar-mass secondary metabolites exhibiting different biological activities, which are used. e.g., in medicine as antimicrobial and antifungal agents, alkaloids and toxins. Some of these substances have highly diverse biological activities and unusual structures. They are produced by streptomycetes, fungi, and bacilli, but interesting products have also been obtained from microorganisms growing in extreme conditions. Several thousands of microbial products have so far been discovered and many other, which can be potentially useful and/or prospective for human use, can still be in the offing. PMID- 11899466 TI - Multilocus enzyme electrophoresis analysis of Pseudomonas syringae pv. phaseolicola. AB - A multilocus enzyme electrophoresis technique was developed to detect variation in seven enzyme loci among isolates of Pseudomonas syringae pv. phaseolicola, representing three races from different geographical locations, the causal agent of the halo blight disease of beans. Cellulose acetate gel electrophoresis of seven enzymes revealed 19 electrotypes (ET) among 21 Pseudomonas syringae pv. phaseolicola isolates. One of the pathovar syringae and one of the pathovar tomato isolates were represented by two different ET. The population of Turkish isolates and three races of the pathovar phaseolicola appeared to be genetically diverse. PMID- 11899467 TI - Oxidation of lincomycin by hydrogen peroxide restricts its potential biotransformation with haloperoxidases. AB - Lincomycin biotransformation was conducted by using Streptomyces venezuelae and Streptomyces phaeochromogenes cell-free extracts. Reaction products were isolated and identified by MS and NMR spectroscopy as lincomycin sulfoxide and lincomycin sulfone. Both compounds arise also by chemical oxidation with hydrogen peroxide; this reaction represents a new efficient way for the preparation of lincomycin sulfoxide and lincomycin sulfone and simultaneously excludes the biotransformation of lincomycin using haloperoxidases. PMID- 11899468 TI - Copper complexes with bioactive ligands Part I--Antimicrobial activity. AB - Biological properties of new copper(II) complexes of 2-methylthionicotinate (2 MeSNic) of composition Cu(2-MeSNic)2(MeNia)(2).4H2O (where MeNia is N-methyl nicotinamide), Cu(2-MeSNic)2(Nia)(2).2H2O (where Nia is nicotinamide) and Cu(2 MeSNic)2L2 (where L is isonicotinamide (iNia) or ethyl nicotinate (EtNic)) are reported. Gram(-)-bacteria (Escherichia coli) are more resistant against Cu(II) complexes than Gram(+)-bacteria (Staphylococcus aureus)--significant antistaphylococcal activity was found with Cu(2-MeSNic)2(MeNia)(2).4H2O (IC50 1.3 mmol/L). Candida parapsilosis was most inhibited by Cu(2-MeSNic)2.H2O and Cu(2 MeSNic)2(MeNia)(2).4H2O (IC50 1.4 mmol/L and 1.5 mmol/L, respectively). Biosynthesis of nucleic acids influenced by Cu(2-MeSNic)2(Nia)(2).2H2O indicated by incorporation of 14C-adenine (IC50(Ade) 0.31 mmol/L) is more sensitive than biosynthesis of proteins indicated by incorporation of 14C-leucine (IC50(Leu) 9.94 mmol/L). Cu(II) complexes with expressed antimicrobial activity showed no mutagenic activity. PMID- 11899469 TI - Application of capillary zone electrophoresis to study the properties of rhodanese from Acidithiobacillus ferrooxidans. AB - A new capillary zone electrophoretic method was applied to the assay of enzymic activity of rhodanese from Acidithiobacillus ferroxidans. The enzyme activity determined by capillary zone electrophoresis was compared with that determined by discontinuous spectrophotometry, the values obtained being in good agreement. The method was also used to evaluate Michaelis constants of cyanide and thiocyanate as substrates; a new approach was developed to solve the problem with variable ionic strength of the samples. The pH and temperature optima for the enzyme were also determined. PMID- 11899470 TI - Subcellular shifts of trimeric G-proteins following activation of baker's yeast by glucose. AB - Addition of glucose to a resting cell suspension of the yeast Saccharomyces cerevisiae was accompanied by marked shifts of the G alpha-protein subunits from the plasma membrane to the cell interior. This process was rapid with half-times between < 10 and 20 s. The decrease of the plasma membrane pool of the Gi alpha/Go alpha- and Gq alpha/Gl 1 alpha-protein subunits correlated with an increase in acid-sensitive forms of these proteins which was recovered in the mitochondrial and/or lysosomal membrane fraction. In contrast to cells from higher organisms glucose-stimulated yeast exhibits an extremely rapid type of the redistribution (internalization). The question remains open as to the functional significance of the internalized forms of the G-proteins as these remain sequestered from the plasma membrane well after glucose has been consumed. PMID- 11899471 TI - Yeast strains from Livingston Island, Antarctica. AB - Five yeast strains were isolated from soil and moss samples from the Livingston Island (Antarctica) and identified according to morphological cultural and physiological characteristics. All strains had an optimum growth temperature of 15 degrees C: none grew above 25 degrees C. They assimilated D-glucose, D galactose, sucrose, cellobiose, trehalose, 2-keto-D-gluconate, D-xylose, D-ribose and melezitose. Four of them were nonfermentative, only one, which formed pseudomycelium fermented glucose, galactose, trehalose. Two strains were identified as pink-red yeasts belonging to genus Rhodotorula--R. minuta and R. mucilaginosa; two were related to the genus Cryptococcus--C. albidus and C. laurentii; one was Candida oleophila. PMID- 11899472 TI - Lipid composition of some yeast strains from Livingston Island, Antarctica. AB - Yeast strains Cryptococcus albidus, Cryptococcus laurentii, Rhodotorula minuta were isolated from a moss sample. Candida oleophila and Rhodotorula mucilaginosa were isolated from a soil sample taken from Livingston Island. Antarctica. Fatty acid, phospholipid, sterol and tocopherol composition was determined in separated lipid fraction after fermentation in a medium containing glucose, peptone and yeast extract. Unsaturated fatty acids, mainly oleic (51-65%) and linoleic (9.5 16.8%), predominated in triacylglycerols. Sterols represent ca. 120-930 mg per kg dry biomass. The content of major phospholipids (phosphatidylcholine, phosphatidylinositol and phosphatidylethanolamine) was ca. 100-800 mg/kg. The amount of tocopherols (mainly gamma- and delta-tocopherol) was 2.1-6.3 mg/kg. PMID- 11899473 TI - Effect of osmotic stress on Aspergillus chevalieri respiratory system. AB - The osmotolerant fungus Aspergillus chevalieri tolerates up to 80% sucrose concentration in the growth medium. At 50% sucrose the growth rate is 1.5-fold higher than in control (3% sucrose), at 80% sucrose it drops to 30% of the control level. Total proteins and lipids in the mitochondrial fractions obtained from the mycelium rise with increasing sucrose concentration during growth (2.6 and 2.1 times at 80% sucrose). The rate of respiration by whole cells and mitochondrial fractions increases with increased sucrose level in the growth medium. The activity of respiratory enzymes, such as succinate dehydrogenase, cytochrome oxidase, NADH oxidase and succinate oxidase, were also higher in cells grown in the presence of elevated sucrose concentrations. The largest increase was observed with NADH dehydrogenase. A. chevalieri cells grown at high osmotic stress exhibited enlarged mitochondria. The mean mitochondrial diameter at 50 and 80% sucrose was approximately 2.9- and 2.6-fold larger than in the control, respectively. Agarose gel electrophoresis of nucleic acids revealed the presence of high-density bands of RNA in mitochondrial fractions from cells grown at elevated sucrose levels. PMID- 11899474 TI - Some nutritional factors influencing mevinolin production by Aspergillus terreus strain. AB - Aspergillus terreus produces the hypocholesterolemic compound mevinolin. Its growth and mevinolin production were affected by the composition of the culture medium. Both were at a maximum with glucose (6%) as the sole carbon source and in the presence of a mixture of yeast extract and sodium nitrate as nitrogen source. Influence of the concentration of some inorganic salts are also discussed. PMID- 11899476 TI - Intraspecific invariability of the internal transcribed spacer region of rDNA of the truffle Terfezia terfezioides in Europe. AB - ITS regions (internal transcribed spacers--ITS1 and ITS2--with the 5.8S gene of the nuclear rDNA) of 25 fruit body samples of Terfezia terfezioides, originating from Hungary and Italy, were compared. The amplification and sequencing of the ITS region was successful with both the ITS1-ITS4 and ITS1F-ITS4 primer pairs. No differences of the restriction fragment length polymorphism profiles were detected among 19 samples collected in one place at the same time. The sequences of the ITS region of 9 samples collected in different localities were highly invariable, differing in only two bases. Thus the intraspecific homogeneity of the ITS region seems to be an important species-specific characteristic of T. terfezioides in contrast to other Terfezia species. As the samples of the species were collected from different and distant localities of Europe, the ITS sequence of T. terfezioides can be considered a very conservative, reliable molecular marker of the fungus. PMID- 11899475 TI - Vegetative growth, aging- and light-induced conidiation of Trichoderma viride cultivated on different carbon sources. AB - The growth and conidiation of the aged Trichoderma viride culture grown in the dark, and after an induction by a light pulse, was examined in the presence of selected mono-, di(tri)saccharides, amino acids and alcohols as sole carbon sources. Hexoses and disaccharides, but not pentoses and amino acids, promoted proportionally both growth and conidiation induced by aging or light. All compounds but pentoses promoted the conidiation in aged cultures and photoconidiation in a close correlation. Ethanol, glycerol and ethylene glycol supported both growth and conidiation but these processes were not supported equally. Conidia formation with hexoses and amino acids as sole carbon sources seems to be a function of growth promotion, rather than of growth restriction (starvation, stress, aging). With glucose as sole carbon source the conidiation was not triggered by nutrient limitation, nor by the accumulation of waste metabolites. The aging-induced conidiation can be considered to be triggered by the genetic program of the microorganism rather than by its nutrient status. PMID- 11899477 TI - D-amino-acid oxidase--an improved production of the enzyme by the yeast Trigonopsis variabilis in a laboratory fermentor. AB - The cultivation of the yeast Trigonopsis variabilis producing D-amino-acid oxidase (an enzyme participating in the transformation of cephalosporin C into 7 aminocephalosporanic acid for the production of beta-lactam antibiotics) was controlled by changes of dissolved oxygen tension and extended fermentation times. The production technology was optimized on a laboratory scale and scale-up parameters were identified. PMID- 11899478 TI - Plastid state- and light-dependent regulation of the expression of nucleus encoded genes for chloroplast proteins in the flagellate Euglena gracilis. AB - Interorganellar regulatory interactions in the flagellate Euglena gracilis were shown to be more complicated than in green algae and higher plants. Euglena plastids have a much more complex influence on nuclear gene expression than was previously thought. The petJ gene for cytochrome c6 represents a group of nucleus encoded genes for chloroplast proteins, the expression of which is influenced by the state of plastids at the transcriptional level. Moroever, the regulation of these genes might be light-dependent. In contrast, for nucleus-encoded small subunit of ribulose-bisphosphate carboxylase, chlorophyll a/b binding protein, and porphobilinogen deaminase transcript levels are unchanged in wild-type cells relative to white mutants. In these cases there is no plastid-derived signal operative during transcription. Porphobilinogen deaminase appeared to be regulated even at the post-translational level. PMID- 11899479 TI - Candida-associated denture stomatitis. AB - Candida-associated denture stomatitis was demonstrated by its cultivation in 171 out of 240 patients examined with partial or total dentures. After taking smears from lesions of the oral mucosa (tongue, cheeks, palate) and the contiguous denture surface by cotton wool swabs and inoculating them onto Sabouraud glucose agar and CHROMagar Candida, individual yeast species were identified by a germ tube, filamentous, and assimilation tests employing the commercial kit AuxaColor. Seven Candida species were identified in smears from the oral mucosa lesions and the contiguous denture surface: C. albicans (95 patients), C. tropicalis (26), C. parapsilosis (20), C. krusei (14), C. guilliermondii (12), C. lusitaniae (1) and C. freyschusii (1). Diabetes mellitus, neoplastic diseases, chemotherapy, radiotherapy, and broad-spectrum antibiotic therapy were identified as some of the large number of factors predisposing patients to stomatitis prothetica. PMID- 11899480 TI - The histopathological characterization of oral Candida leukoplakias. AB - Sixty four biopsy specimens of oral Candida leukoplakias were examined. Histological sections were stained with periodic acid-Schiff (PAS) reagent and by Grocott's silver method. C. albicans isolates were cultivated from all the patients but fungal hyphae were proved histologically only in 23 of them. PAS positive and Grocott-positive elements were exclusively observed in the superficial epithelial layers, e.g., the cornified layer and the stratum granulosum. The inflammatory reaction was characterized by an early phase during which polymorphonuclear cells predominated. The skeletal muscle fascicles immediately adjacent to the infected epithelium showed striking degeneration and atrophy associated with a marked infiltrate of chronic inflammatory cells. PMID- 11899481 TI - Electrophoretic karyotyping of Candida albicans strains isolated from premature infants and hospital personnel in a neonatal intensive care unit. AB - Electrophoretic karyotyping was used to compare DNA probes of yeasts isolated from blood of preterm neonates (n = 66) in a neonatal intensive care unit (NICU) and from the hands of healthy hospital personnel (n = 10). The yeasts were identified as Candida albicans using standard laboratory methods. DNA was extracted from yeasts and isolation of identical DNA strains from the pairs nurse neonate suggested that one nurse transmitted one yeast strain by her hands to three neonates. Four neonates harbored two identical strains originating from two nurses, i.e. each nurse transmitted the same strain to two neonates. In the additional 7 cases transmission of 1 yeast strain by 1 nurse to 1 neonate was observed. Our data suggest that nonperinatal nosocomial transmission of C. albicans occurs in neonates, possibly via cross-contamination being transferred on hands of health care workers. The importance of careful hand washing of staff (health care workers) and other infection-control procedures (to prevent the nosocomial transmission of pathogens in the NICU environment) is emphasized. PMID- 11899482 TI - Effect of low and high doses of Salmonella enteritidis PT4 on experimentally infected chicks. AB - Chicks (1-d-old, three groups, each containing 50 chicks) were inoculated with 2 x 10(2) and 2 x 10(8) CFU of Salmonella enteritidis; the third group were kept as uninoculated control. Five birds from each group were euthanized at intervals from 6 h to 4 weeks post-inoculation (pi). In the low-dose group S. enteritidis was isolated from 60% cecal samples at 18 h pi, and from 20% of livers at 3 d pi. Individual variation in the frequency of S. enteritidis recovery was observed in this group. The clearance of salmonella from the organs was faster in the low dose group, and salmonella was not isolated from the liver and cecum at 21 and at 27 d pi, respectively. However, in the high-dose group, S. enteritidis was isolated from all ceca and 80% of liver 6 h pi, and salmonella was detected in the cecum and liver throughout the experiment. Serous typhlitis and unabsorbed yolk sac were the most prevalent lesions in both groups. Granulomatous nodules in the cecum were found occasionally in some cases in both inoculated groups, which can play a role as reservoirs in carrier chicks. PMID- 11899483 TI - [Treatment of insomnia in demented nursing home patients: a review]. AB - Insomnia and fragmentation are features of the sleep of these patients. In order to list the factors disturbing the sleep of demented nursing home patients and the interventions improving their sleep quality, the literature was reviewed. A Medline search over the period 1966-2000 was performed. This resulted in 22 research articles. Admission to a nursing home is associated with sleep disturbances caused by patient problems (e.g. pain), care routines (e.g. nightly nursing round) and environment (e.g. noise). There are indications that the use of hypnotics in nursing home patients is not always effective and increases the risk of falls. There are several ways to reduce hypnotic consumption in nursing homes. Non-pharmacological interventions to decrease sleep disturbances caused by environmental factors have a favourable although weak effect on sleep itself. By reducing nightly noise, sleep quality does not necessarily improve. Light therapy seems to be the most effective non-pharmacological method to strengthen the circadian sleep/wake rhythm. The struggle against insomnia without using medication perhaps requires a two tracks management: detection and elimination of disturbing environmental factors and implementation of an adequate method to strengthen the circadian sleep/wake rhythm. PMID- 11899484 TI - [Risk profiles and preventive measures of falls in elderly persons]. AB - Falls in elderly persons are an important health problem. The results of the Longitudinal Aging Study Amsterdam show that thirty percent of older adults over the age of 65 years who live in the community (n = 1285) fall at least once a year. Recurrent falls were reported by about 11% of the participants. In one-year of follow-up, 22 fractures were recorded. In the 'single fall' group 11 subjects (3.9%) suffered from a fracture and in the 'recurrent fall' group 9 subjects (6.1%). The strongest predictors identified in the risk profile for recurrent falls were previous falls, urinary incontinence, visual impairment and functional limitations (Area Under Curve, 0.71). The probability of recurrent falls for subsequent scores of the screening test ranged from 4.7% (95% CI, 4.0-5.4%) to 46.8% (95% CI, 43.0-50.6%). Risk profiles are needed to identify people at high risk. For matters of feasibility and efficiency, preventive measures of falls should preferably be focussed on those subgroups that have the highest risk of falls. PMID- 11899485 TI - [Pensions and becoming older. An economic analysis of fertility, migration and pension legislation]. PMID- 11899486 TI - [Diagnosis of personality disorders]. PMID- 11899487 TI - [Elderly: the image in the public eye. The 'luxury lives of the elderly' versus the bitter truth]. PMID- 11899489 TI - [Ethical dilemmas in early detection of dementia. An inventory of moral intuitions in practice]. AB - In this article we present an inventory of the moral intuitions of the health care workers who work in the field of early detection of dementia. The effects of pharmacological treatment and professional care and support may improve when dementia is detected in an early stage. Furthermore, the patient (and his family) can prepare themselves for the period to come. Health care workers recognize moral problems and tensions concerning early detection that are related to the question whether persons will benefit from knowledge in an earlier stage of dementia, because this knowledge can be a heavy burden. We asked general practitioners, home care workers, employees of the so-called 'Memory clinic' and specialists, what ethical intuitions they recognize in practice. They mentioned the following questions: when are health care professionals allowed to take initiative, is causing worries and concerns problematic, and should a diagnosis always to be told? We conclude after a first analysis that many moral questions derive from the fact that many health care professionals lack knowledge of the wishes and interests of the elderly. At the same time they try to justify their actions on the (presupposed) consent of the elderly person. We suggest that the general norm 'only act when the patient wants to be helped' in health care should also apply to detection of dementia, although it should not be taken too strictly. Another justification for early detection can be found in the benefits for the elderly people, when their wishes are no longer expressed. PMID- 11899490 TI - [From doubts born after September 11 to hope placed in the euro...]. PMID- 11899488 TI - [New structures in geriatric care: the geriatric home consultation by a nurse practitioner]. AB - The growing number of elderly and chronically ill people causes an increasing demand for care. New patterns in care for geriatric patients are required, to guarantee geriatric care in the future. In the Transmural Model for Geriatric Care, the geriatric nurse practitioner participates in geriatric home consultation. The geriatric nurse practitioner makes the home visits of the geriatrician. First experiences with home consultation by geriatric nurse practitioner are positive. The input of the geriatric nurse practitioner in home consultation has two goals: care substitution and improvement of quality of care. Substitution of care enlarges the possibilities of the geriatrician, which are limited now, because of the enormous demand for geriatric care. The specific tasks of the geriatric nurse practitioner are functional assessment and care coordination. PMID- 11899492 TI - [Generic drugs]. AB - Various incentives will progressively increase the part of generic drugs on the Belgian market place and will more and more confront the prescriber with the problem of substitution. The equivalence between generic and brand name products and the fear, justified or not, at efficacy or patient safety levels in case of substitution are discussed on the basis of biopharmacy and pharmacokinetic concepts in connection with modalities generally adopted when requesting marketing authorisation. Considering the extent of their utilisation at international level and the low frequency of clinically significant problems linked to bioequivalency, limited to a small number of substances, prescribers should be soon released from their apprehensions on the quality of generic drugs. On another hand, they must be stimulated to be vigilant when they substitute a drug by any other one, especially in the cases of drugs, classes of drugs, or pharmaceutical forms considered as critical and when occurring to patients whose disease or physiological status are able to amplify bioequivalence problems. PMID- 11899491 TI - [Primary linitis plastica of the rectum]. AB - One case of primary linitis plastica of the rectum is presented, it is a rare entity presentation in younger patients. The diagnosis was made by excluding the stomach as a primary source by peroperative palpation and fibroscopy examination. Because of potential for local invasion and infiltration, linitis is often detected only at the equivalent of stage C (Astler-Cller). The prognosis is poor, the mean survival time is about 9 months. PMID- 11899493 TI - [Morphological and metabolic abnormalities related to antiretroviral therapy]. AB - The long term use of antiretroviral therapy is associated with metabolic and morphological abnormalities named lipodystrophy. The understanding of its epidemiology is in progress. NRTIs and Pls are involved in the origin of these abnormalities. So, NRTIs inhibit mitochondrial gamma polymerase and induce body fat distribution disorders. PIs interfere with lipid metabolism, leading to hyperlipidaemia and insulin resistance symptomatology. The diagnosis is made by clinical (far wasting or accumulation, or mixed syndrome) and biological signs (hyperlipidaemia, hyperglycaemia, hyperlactacidaemia). Radiological exams can quantify morphological abnormalities. The management consists in drug substitution, diet and exercise, and corrective drugs. Plastic surgery can be useful. PMID- 11899494 TI - [Tanning benches to accusation benches]. AB - Sunbeds used in sun parlours do not garantee a controlled innocuity. By contrast, health hazards are obvious in many instances. Forceful tanning is a transitory aspect behind which the spectrum of many skin changes includes accelerated photoageing, irreversible pigmentary and keratotic changes, and cutaneous malignancies including malignant melanoma. In the absence of protection, the eye is also affected. Even if tanning salon exposure increases the risk for skin cancer by a small amount, it represents a significant potential public health hazard given the number of people attending salons. Digital dermoscopy and ultraviolet light videoscopy are new methods allowing to predict the cutaneous ransom to be paid by the sunbed worshippers. PMID- 11899495 TI - [How I treat ... facial paralysis by hypoglosso-facial anastomosis]. AB - Thirteen patients underwent a hypoglosso- or a spino-facial nerve anastomosis between 1990 and 1996. Facial palsy was the result of surgery in 12 cases and of radiosurgery in 1 case. The mean interval between facial palsy and anastomosis was 12 months. Facial nerve function is determined on the basis of clinical examination according to the classification of House-Brackmann and our own evaluation. According to House, 10 patients are classified grade III and 3 grade IV. Our evaluation defines in 10 grade III, 7 good results and 3 fair results. As far as the good results are concerned, the mean interval between palsy and anastomosis is short (< 3 months). The permanent eating and swallowing dysfunctions are consecutive to multiple cranial nerve deficits. The post paralysis hemifacial spasm is facilitated by prolongated electric stimulations. PMID- 11899496 TI - [Hemolytic uremic syndrome and thrombotic thrombocytopenic purpura]. AB - Hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP) are thrombotic microangiopathies (TMA). Both familial and sporadic forms exist: anaemia, thrombopaenia, renal failure and neurologic disorders are common clinical features. The differential diagnosis depends on plasma levels of von Willebrand factor-cleaving protease: there is a deficiency of this protease in patients with TTP whereas its rate is normal in HUS. We remind pathophysiology, etiologies and treatment of these TMA. PMID- 11899497 TI - [Dog bite in a splenectomized patient]. AB - We present a case of Capnocytophaga canimorsus fulminant infection linked to a dog bite in a splenectomized patient. Capnocytophaga canimorsus is a gram negative rod that typically causes septicaemia with disseminated intravascular coagulation in both immunocompromised and immunocompetent hosts. It is associated with high mortality. We also reviewed the literature and provide some recommendations on the management of bite wound as well as on both prevention and treatment of infection in asplenic state. PMID- 11899498 TI - [Pulmonary bronchogenic cysts: clinical observation and literature review]. AB - Pulmonary bronchogenic cysts are benign lesions that can be suspected from clinical background and imaging. We present the case of a huge subcarinal bronchogenic cyst and review the embryology, physiopathology, surgical indications and techniques of this congenital lesion. PMID- 11899499 TI - [War and medicine in a culture of peace. 3. Synopsis of chemical warfare agents]. AB - A variety of chemical components can be used as warfare threats directly targeting humans. They can be classified according to their main biological effects as nerve agents, vesicants, lung damaging agents, cyanogen agents and incapacitants. Other chemical agents are water and food contaminants. Still other less aggressive compounds are mainly used to control riots. Smokes, flame materials and herbicides belong to other classes of chemical agents of the warlike armamentum exhibiting a direct effect on man. PMID- 11899500 TI - [Pharma-clinics medication of the month. Rabeprazole (Pariet)]. AB - Proton pomp inhibitors (PPI) have revolutionized the treatment of gastro oesophageal reflux disease and gastro-duodenal ulcers. Rabeprazole (Pariet) is the last PPI arrived on the Belgian market. Controlled studies have shown an efficacy similar to emeprazole in the treatment of oeso-gastro-duodenal acid diseases. Rabeprazole has a favourable profile with rapid action and good safety. PMID- 11899501 TI - [Clinical study of the month. Effects of valsartan in chronic heart failure: the VAL-HeFT study]. AB - Current guidelines recommend drugs which reduce neurohormonal activation as standard therapy for heart failure: angiotensin converting-enzyme (ACE) inhibitors, beta-blockers and spironolactone. The Valsartan Heart Failure Trial (Val-HeFT) tested the efficacy of the angiotensin-receptor blocker valsartan in addition to prescribed therapy. A total of 5010 patients with heart failure of NYHA class II, III or IV were randomly assigned to receive 160 mg of valsartan or placebo twice daily. Further angiotensin antagonism by valsartan did not improve survival but was beneficial in terms of morbidity and mortality, because of reduced rate of hospitalization, significant improvements in NYHA class, ejection fraction, signs and symptoms of heart failure, and quality of life. Valsartan had highly favourable effects in patients not receiving ACE inhibitors but an adverse effect in patients receiving both ACE inhibitors and beta-blockers. The effects of adding valsartan are depending on the importance of previous neurohormonal inhibition. PMID- 11899502 TI - [Clinical case of the month. Subvalvular aortic stenosis]. AB - A case of fixed subvalvular aortic stenosis is reported. This clinical observation is the occasion to review the pathophysiology of this lesion and the last developments in the surgical treatment. PMID- 11899503 TI - [Experience of pain during childbirth]. PMID- 11899504 TI - [Diagnosis of infectious disease. Actinomycosis]. PMID- 11899505 TI - [The use of albumin infusion in decompensated liver cirrhosis]. AB - For patients with refractory ascites, paracentesis is the standard therapy and for many it is the only treatment option. When more than five litres of ascitic fluid are removed, the use of a plasma expander effectively prevents "postparacentesis circulatory dysfunction", which is associated with a high mortality. Randomised controlled studies show that albumin is more effective than synthetic plasma expanders in the prevention of this complication. In selected patients with ascites, long-term administration of albumin may improve the diuretic response. A randomised controlled study in patients with spontaneous bacterial peritonitis has demonstrated that treatment with albumin infusion in addition to an antibiotic reduces the incidence of hepatorenal syndrome. Albumin infusion in combination with the administration of a vasopressin analogue may be able to reverse established hepatorenal syndrome; however, no controlled studies have been published. Whereas the use of albumin infusion with large-volume paracentesis is strongly supported by the available evidence, additional conclusive studies of the use of albumin for spontaneous bacterial peritonitis are awaited. PMID- 11899506 TI - [Is intravenous albumin relevant in the treatment of patients with liver disease?]. PMID- 11899507 TI - [Albumin in decompensated liver cirrhosis]. PMID- 11899509 TI - [The anterior cruciate ligament]. AB - The anterior cruciate ligament (ACL) has a mechanical and a sensory function during daily activities and during strenuous activity, such as sport. There are 2,500 ACL ruptures/year in Denmark, mostly occurring during sports. Usually the diagnosis can be established clinically. Acute suture of the ligament gives unsatisfactory results, but reconstruction provides mechanical stability and good knee function in 80%. Less than 50% are able to return to top level sports. The long-term risk of degenerative changes in the knee is increased 10-15 fold after an isolated ACL rupture. Untreated ACL ruptures have a particularly poor prognosis in children. Proprioceptive training can reduce the risk of ACL injuries in football. There is little evidence in support of treatment strategies with respect to surgery and rehabilitation after ACL injury. Randomized studies should be initiated, and in Denmark epidemiological data on ACL injuries and treatment could be registered in a national database. PMID- 11899508 TI - [Ingestion of button batteries. Epidemiology, clinical signs and therapeutic recommendations]. AB - The frequency of ingested button batteries is about 10 per million population per year, and one in every 1,000 battery ingestions causes serious injuries. Most of the patients are children. Seven to 20% develop transient symptoms, owing to irritation or superficial corrosion of the gastrointestinal tract. The severe lesions are almost always confined to the oesophagus. Toxic effects from the battery contents are unimportant. Batteries lodged in the oesophagus may not cause initial symptoms, and it is recommended that when ingestion is suspected patients should be examined by fluoroscopy. Batteries in the oesophagus should be removed promptly by endoscopy, those distal to the oesophagus can be left to pass spontaneously. Passage can be ensured by examination of the stools. PMID- 11899510 TI - [Experience of pain in relation to birth weight in primiparae]. AB - INTRODUCTION: The aim of the study was to investigate the association between birth weight and the labour pain experienced by primiparae. METHODS: All primiparae who gave birth at the Department of Obstetrics, Herning Central Hospital, from 1 September 1998 to 30 April 1999 completed a visual analogue scale on the second day after delivery. Pain was scored in the first, second, and "repair" stage of labour, and lastly a score was assigned for overall assessment of labour. RESULTS: A total of 139 primiparae were entered. The mean age was 27 (range 16-40) years and the mean birth weight was 3562 (range 2400-5050) grams. There was no relation between birth weight and pain score. Nor was there any relation between pain score and maternal age, body mass index, duration of the second stage of labour, or the need for instrumental delivery. The score in the stage of perineal repair was significantly lower than in the other stages of labour. CONCLUSION: The pain experienced during labour in primiparae is not influenced by birth weight. PMID- 11899511 TI - [Reconstruction of the anterior cruciate ligament in patients over 40 years of age]. AB - INTRODUCTION: The treatment of an ACL lesion in patients who are above 40 years of age remains controversial. The aim of this study is to report the subjective impression of the result after ACL reconstruction in 24 such patients. MATERIAL AND METHODS: The records of 24 patients, who were aged 40 or older when they had ACL reconstruction were reviewed. The average age was 43 years (40-49), 10 women and 14 men. The mean interval between injury and operation was 35 months (6-185). All 24 patients completed a self-administered questionnaire to evaluate A) preoperative status, B) the rehabilitation period, and C) postoperative status and their overall view of the treatment. Status was made 16 months postoperatively (5-41). RESULTS: Preoperatively, 23 out of 24 patients had instability problems in daily living (ADL) and at work. One had not experienced the knee giving way. All the patients had experienced pain. Postoperatively, 21 out of the 24 experienced no instability and 14 are free of pain. Eighteen have increased activity, five have no change in activity, and one has decreased activity. Sixteen are satisfied with the level of postoperative activity. Twenty two find their knee status better postoperatively than preoperatively. One finds it unchanged, and one finds it worse. Overall 15 patients find result very satisfying, seven find it satisfying, and two find it poor. Twenty-two would advise others in a similar situation to have the operation done, whereas one is in doubt and one would not recommend the operation. DISCUSSION: We conclude that the results of ACL reconstruction in patients above the age of 40 are as good as those in younger patients. PMID- 11899512 TI - [Evaluation by pregnant women of prenatal counseling. A quality assurance project]. PMID- 11899513 TI - [Laparoscopic surgery of Spiegelian hernia]. AB - A case of spigelian hernia in a 33-year-old woman, diagnosed preoperatively and operated on laparoscopically, is described. Simultaneously, a bilateral femoral hernia without previous symptoms was diagnosed and treated. Goretex dualmesh was used for the spigelian hernia, polypropylene plugs for the femoral hernias, fixation with Goretex sutures and metal coils. Laparoscopic approach for the diagnosis and treatment of rare hernias in the abdominal wall is advocated, because of the diagnostic potential and reduced postoperative discomfort. PMID- 11899514 TI - [Picture of the month: perforated ventricular ulcer]. PMID- 11899516 TI - ["Crisis intervention"]. PMID- 11899515 TI - [Patient survival after human albumin administration]. PMID- 11899517 TI - [Diagnostic knee arthroscopy under local anesthesia in hospital]. PMID- 11899518 TI - [The debate on glitazones]. PMID- 11899519 TI - [Diagnosis of malignant trophoblastic disease in women]. PMID- 11899520 TI - [Levels of lipid peroxidation products and ceruloplasmin in blood as characteristics of tolerance to physical load in hypertrophic cardiomyopathy]. AB - The effect of standard bicycle ergometry test on the serum concentrations of lipid peroxidation (LPO) products and ceruloplasmin were studied in healthy volunteers and patients with hypertrophic cardiomyopathy. The initial levels of LPO products and ceruloplasmin in the patients were higher than in volunteers. In volunteers bicycle ergometry test did not affect the content of ceruloplasmin in the blood and led to accumulation of circulating LPO products. In patients with hypertrophic cardiomyopathy bicycle ergometry did not lead to increase of lipoperoxide level and promoted an increase in the blood ceruloplasmin concentration. PMID- 11899521 TI - [Identification of cysteine-containing proteins following separation by disk electrophoresis]. AB - Several approaches to measurements of cysteine-containing proteins are known. These proteins are essential for normal function of cells, tissues, organs, and the total organism under natural conditions and during exposure to exo- and endogenic pathological factors. Unfortunately, the routine methods are inconvenient and have many drawbacks, which impedes their wide use. The authors suggest a simple, cheap, and rapid method for identification of cysteine containing proteins after their separation by disk electrophoresis in polyacrylamide gel. PMID- 11899522 TI - [Food sensitization and recurrence frequency in patients with chronic gastric erosions]. AB - Besides well-known pathogenetic factors of chronic gastric erosions (CGE), sensitization to alimentary, communal, and infectious allergens and antigens contributes to development of relapses of this condition. Moderately high and high levels of specific IgE and significant increases in the total IgE level were detected in patients with CGE relapses in comparison with the controls. Patients with CGE develop secondary immunodeficiency, which was confirmed by insufficient production of serum IgA during disease exacerbations. The sensitization status in CGE patients persists during remissions It is characterized by moderate levels of specific IgE, indicating chronic transformation of the process. PMID- 11899523 TI - [Purification of antibodies for fluorescent diagnosis in influenza]. PMID- 11899524 TI - [Clinical evaluation of phagocyte tests in urogenital chlamydiosis]. PMID- 11899525 TI - [24-Hour intragastric pH measurement in a hospital for internal diseases]. PMID- 11899526 TI - [Hematological characteristics in blood autoanalysis in children with congenital heart defects]. AB - Hematological status of infants with congenital heart disease (CHD) was evaluated by automated systems. Blood analyses were carried out in 40 infants with the pale type and in 20 with the blue type CHD. Increased erythrocyte count and hematocrit were detected in the blue type CHD. Evaluation of the leukocytic formula showed correlation of the results of automated analysis and microscopic analysis of a blood smear. Automated method is preferable for estimating platelet count. Platelet count was decreased and the cells were larger in patients with the blue type disease in comparison with the pale type group. Hence, automated analyzer gives a wide spectrum of blood values with high accuracy and productivity and helps evaluate hemopoiesis in patients with CHD. PMID- 11899527 TI - [Determination of creatine kinase MB isoenzyme mass in serum of patients with myocardial infarction]. PMID- 11899528 TI - [Diagnostic significance of chemiluminescent response of neutrophil granulocyte in children with lymphadenopathy]. AB - Chemiluminescent response of the blood neutrophilic granulocytes was evaluated during the diagnostic period in 22 patients with Hodgkin's disease, 35 with nonspecific lymphadenopathies, and 24 patients with tuberculous infection aged 7 14 years. Disorders in spontaneous and induced chemiluminescence of neutrophils were detected in all patients, and were the most pronounced and stable in patients with Hodgkin's disease. Evaluation of neutrophilic chemiluminescent response in these patient groups leads to a more accurate diagnosis of lymphadenopathies of different origin. PMID- 11899529 TI - [Hemorheology and morphofunctional characteristics of blood cells in children with congenital heart defects]. AB - Patients with congenital heart diseases (CHD) aged 4 months to 12 years, hospitalized for surgical treatment, were examined. Complex viscosity of the blood and the constituents of this parameter (dynamic viscosity and elastic component) were evaluated in the range of shift strain corresponding to the range of shift velocities 0.37-500 s-1 at a frequency of 2 Hz. The threshold blood fluidity was estimated for evaluating the conditions under which erythrocyte disaggregation begins. The relationship between blood rheology and morphofunctional characteristics of blood cells was evaluated. In accordance with the findings, the patients were divided into 3 groups with different suspension stability of the blood. In group 1 the threshold fluidity approximated the norm. In group 2 with normal blood rheology the shifts were compensated and regulation of rheology was in general intact. In group 3 the hematocrit values, mean erythrocyte volume, mean concentration of hemoglobin in erythrocyte, leukocyte counts, and complex viscosity of the blood were the highest, while the suspension stability was the lowest, which indicates depletion of the adaptive potential of the organism. Hence, blood rheology in patients with CHD differs by the type of regulation, mechanisms and compensation of changes, and depend largely on the mean erythrocyte volume, mean hemoglobin concentration in erythrocytes, and leukocyte counts. PMID- 11899530 TI - [Analytical method for evaluation of coagulation activity of factor VIII]. AB - A single-stage method is used for evaluating factor VIII (fVIII) clotting activity. This method is based on evaluation of activated partial thromboplastin time with fVIII-deficient plasma as the substrate. The activity of fVIII in the plasma is estimated by the graphic analytical method, which is difficult and not sufficiently accurate. An analytical method for estimating the activity of fVIII is validated. The authors prove the adequacy of empirical model used for evaluation of fVIII activity on the basis of results of processing of a vast scope of experimental data. The range of values for this parameter is -6.00 +/- 0.16. PMID- 11899531 TI - [Effect of activators for activated partial thromboplastin time test on its results in the tube and automated test versions]. AB - Effects of activators used in combination with partial thromboplastin (PT), manufactured by the Kirov Institute of Hematology and Blood Transfusion, on the results of the activated partial thromboplastin time (APTT) test and its sensitivity to heparin were studied. Kaolin manufactured in Russia and by Aldrich firm and silica from Sigma were used. The method was reproduced in tubes and on Organon Teknika photooptic coagulometers: semiautomatic Coag-A-Mate XM and automated Coag-A-Mate RA-4. The concentration and optical density of the activator affected the results of APTT test, which prompts the use of kits calibrated for the PT/activator combination but not separate reagents. Combination of PT manufactured by the Kirov Institute and suspension of Russian kaolin ensured high sensitivity of the method to heparin content in the plasma and good reproducibility of the method both in tubes and on Organon Teknika coagulometers: coefficient of variation of APTT in all variants of the tube test were no higher than 10% and in the automated variant 2.6-6.2%. PMID- 11899532 TI - [The impression smear from the nasal mucosa in diagnosis of respiratory diseases]. PMID- 11899533 TI - [Modification of the Bilshovsky method for studies of nerve tissue paraffin sections]. AB - The Bilshovsky method is often used for studies of the morphology of neuronal fibrillar system. The classical variant of this method requires sections made with a freezing microtome, which is sometimes inconvenient. Preparation of paraffin sections often gives unsatisfactory results. The authors managed to overcome this deficiency. Their modification consists in the following: after deparaffinization of paraffin sections glued to the slides they are repeatedly fixed in warm formalin and then treated with silver nitrate with heating. This modification is easily reproducible and essentially simplifies staining; no freezing microtome is needed. PMID- 11899534 TI - [Improvement of efficacy of the bacteriological diagnosis in whooping cough]. PMID- 11899535 TI - [Modern clinical laboratory: measure of progress and traditions (rationalism?)]. PMID- 11899536 TI - [Rational equipment for laboratory service for the regional health care system]. PMID- 11899537 TI - [A model of a laboratory]. PMID- 11899538 TI - [Quantitative ultraviolet, visible, and near infrared spectrophotometric analysis]. AB - The potentialities of photometry in the ultraviolet, visible, and infrared spectra are compared. Errors associated with neglect of the Booger-Lambert-Ber law, nonmonochromic light, inconstant length of optic route, and non-optimal choice of optic density range are analyzed. PMID- 11899539 TI - Training of clinicians for public health events relevant to bioterrorism preparedness. PMID- 11899540 TI - [Chromosome analysis in medicine]. AB - Aberrations of the normal number and structure of chromosomes can cause mental, growth and developmental delay, defects of sexual development, congenital defects, abnormal facial features, deformation of extremities, defective intrauterine development, spontaneous miscarriages, and infertility. Expressive changes of the karyotype are used to be found in cancer cells. In majority of cases a lot of genes are abundant or deleted and then many organs are affected- therefore disorders are described as "syndromes". Down and Turner syndromes are the best known syndromes caused by numerical aberrations of human chromosomes. In the diagnostics of structural aberrations, the use of methods of classical cytogenetics becomes limited. Fluorescence in situ hybridisation (FISH) is therefore used more frequently. FISH method has several advantages: rapidity, sensitivity, specificity and possibility to apply this method also in interphase nuclei. Application of FISH allows us to detect submicroscopic deletions and amplifications and to explain the aetiology of inborn developmental defects and cancer diseases (including familiar cases). The newest FISH modifications perform one step analysis of multiple chromosomal rearrangements and help us to ascertain the diagnosis and the prognosis of cancer diseases. The use of special computer software for chromosomal and FISH analysis is the most important part of the current cytogenetic diagnostics. PMID- 11899541 TI - [Chromosome instability syndromes]. AB - We refer 55 cases of the chromosomal instability syndromes (SCI), diagnosed in patients of our genetical clinics. Problems of early diagnosis can be documented by a discrepancy between the expected number of patients and their relative advanced age at the time when SCI was ascertained. We have also shown that NBS patients can be diagnosed earlier and the disease sufficiently confirmed on the basis of congenital microcephaly and on the direct detection of 657de15 mutation in NBS1 gene. Genealogical analysis of families with SCI revealed a low risk of prenatal selection of affected homozygotes and high cancer prevalence in relative (in NBS families recognized heterozygotes) at young adult age. Due to severe DNA repair disorder and hyperradiosensitivity of affected homozygotes as well as unaffected heterozygotes, conventional diagnostics and treatment protocols of lymphoreticular malignancies in affected homozygotes are prohibited. The use of Nijmegen treatment protocol improved in our patients dramatically their clinical prognosis, which is documented by 6 NBS patients surviving one or two malignancies. Early diagnose of SCI and information for families and their doctors about consequences of DNA repair disorder and about their hyperradiosensitivity is essential for improving the clinical prognosis of SCI patients. PMID- 11899542 TI - [Specialized genetic counseling in pediatric and adult oncology patients]. AB - Five to ten percent of oncological diseases exhibit monogenic mode of inheritance. They occur as a consequence of the germline mutations of tumor suppressor genes and of the genes engaged in reparative processes. Most common monogenically determined oncological diseases are: AD form of breast and ovarian cancer, hereditary nonpolyposis colorectal cancer (HNPCC, Lynch sy.) and familiar adenomatous polyposis (FAP). The aim of the genetic investigation is to evaluate whether the index family deals with the hereditary form of tumor predisposition, than, if possible, to perform DNA analysis in the family and to propose preventive screening program (methods) for the probands in risk. PMID- 11899544 TI - [140 years' of the Casopis Lekaru Ceskych (Journal of Czech Physicians)]. PMID- 11899543 TI - [Medical genetics in reproductive medicine]. AB - Reproductive genetics (RG) is another new field of medical genetics, integrated with reproductive medicine, assisted reproduction and developmental genetic. RG is closely linked to the perioconceptional prevention, perinatology, ultrasound and biochemical screening in the end of the first and beginning of the second trimesters. RG is based on the system of specialized genetic counseling, clinical cytogenetics, molecular cytogenetics and molecular genetics to provide prefertilization, preimplantation and classical prenatal diagnosis in the Ist to IIIrd trimesters. Thus, RG is part of the fetal medicine and therapy. The six years experience with RG is summarized. A system of the specialized health care, organized, if possible in one integrated center of RG and reproductive medicine (RM) is presented. Reproductive medicine provides all necessary clinical gynecological and andrological surveillance, with assisted reproduction and further obstetrical ultrasound examinations, including nuchal translucency measurements and 2D, 3D ultrasound, echocardiography examinations, if indicated, as well as the invasive method of prenatal diagnosis and perinatology care. Specialized genetic counseling and cytogenetic analysis, if indicated, should be offered to all partners with reproductive disorders as well as to oocyte donors. Chromosome anomalies are disclosed in 6% of men with abnormal sperm analysis as well as in women with severe reproductive disorders. In males with severe oligo, azoospermia, the sperm aneuploidy analysis by molecular cytogenetic methods is recommended. Advised is also the molecular genetic detection of Y chromosome microdeletions, which is detected in 9% of our azoospermic men with deletions in AZFb region. CFTR gene mutations and intron 8 and 10 polymorphism examination is provided not only in men with obstructive azoospermia (CBAVD), but also if severe oligospermy with less than 1 x 10(6) sperm/ml is detected. Molecular genetic analysis of thrombophilic mutations of factor II., V. (Leiden) and MTHFR gene in unexplained recurrent abortions and in cases with unsuccessful IVF is part of the diagnostic strategy. The population frequencies of carriers of mutations of factor II. (2.3%), factor V.-Leiden (5.7%) and MTHFR gene (38%) were determined. The laser biopsy of the first polar body and of blastomeres was introduced for FISH analysis of chromosome aneuploidies. Quantitative fluorescent PCR (QFPCR) detection is used for testing of the most frequent delta F508 CFTR gene mutation and the most frequent aneuploidies of chromosome 13, 18, 21, X and Y. QFPCR was successfully tested for male fetal sex examination from partially purified fetal cells in the maternal blood. The first trimester ultrasound and biochemical screening is recommended to all successful pregnancies after different IVF methods. If borderline levels of first trimester biochemical screening of PAPP-A protein and beta hCG are detected without pathological ultrasound findings, classical triple test of biochemical screening in 16th week of gestation is recommended. If pathological results of ultrasound and biochemical screening are disclosed, invasive prenatal genetic diagnosis is indicated as well as in pregnancies after ICSL, if there is not any obstetrical contraindication. PMID- 11899545 TI - [The human genome and medicine]. AB - Recent publication of the working draft of the human genome and its first analyses revealed several surprising findings. The human genome contains only about 42,000 genes, contrary to previous estimates of about 100,000. Comparison with other genomes suggests that the complexity of an organism need not result from increasing gene number, but it can be based on regulatory mechanisms associated with alternative gene expression and on complex interactions of genes and/or their protein products. The progress in genomics and related modern disciplines is influencing substantially the biomedical research and the medicine itself, where the main focus shifts towards multifactorial diseases. The new knowledge will lead to much more effective diagnosis, exact prognosis of the disease course and of individual drug response, to the targeted therapy using new drugs and gene therapy, and mainly towards targeted prevention based on the detailed knowledge of individual disease predisposition. PMID- 11899546 TI - Deep leg ulceration. Look to underlying systemic disease for the cause of this dermatologic condition. PMID- 11899547 TI - Acute abdominal pain. Four classifications can guide assessment and management. AB - Abdominal pain is a common occurrence in older persons and a frequent catalyst for office and emergency room visits. Complaints must be investigated thoroughly because they often indicate serious underlying pathology such as Infection, mechanical obstruction, malignancy, biliary disease, cardiac problems, and GI ischemia. One means of overcoming a sprawling differential diagnosis is to determine whether the problem falls into one of four general categories: peritonitis, bowel obstruction, vascular catastrophe, or nonspecific abdominal pain. A comprehensive history, careful physical examination, and use of abdominal imaging studies facilitate effective assessment. As atypical presentations are frequently encountered in older persons, liberal use of ultrasound and contrast CT and early surgical consultation are recommended. PMID- 11899549 TI - Bacterial pneumonia. Managing a deadly complication of influenza in older adults with comorbid disease. AB - In patients with Influenza, the risk of death from pneumonia is closely associated with age and chronic conditions. Mortality from influenza and pneumonia in Americans age > or = 65 has been increasing since 1980. Pneumonia following influenza is usually caused by a secondary bacterial infection. Pathogens most commonly implicated are Streptococcus pneumoniae, Staphylococcus aureus, and Haemophilus influenzae. Prompt empiric therapy effective against the suspected pathogen is indicated, whether the patient is being treated as an outpatient or requires inpatient observation or hospitalization for i.v. administration. Influenza vaccination of older patients living in the community has been shown to decrease hospitalizations for influenza and pneumonia by 52% and mortality by 70% in those with chronic lung disease. Protective rates are similar for residents of long-term care facilities. PMID- 11899548 TI - Parkinson's disease. Therapeutic strategies to improve patient function and quality of life. AB - Idiopathic Parkinson's disease (PD) is an age-related neuro-degenerative disorder characterized by slowness, stiffness, resting tremor, gait impairment, and postural instability. Levodopa is the most potent pharmacologic agent for symptom management and is associated with an increase in quality of life and longevity for patients with PD, but chronic use causes motor complications. The availability of several newer types of agents--dopamine agonists, monoamine oxidase inhibitors, and catechol-O-methyltransferase inhibitors--gives physicians increased flexibility with regard to first-line therapy, adjunct therapy, and managing or reducing the frequency of motor complications and other side effects associated with chronic levodopa therapy. PMID- 11899550 TI - Creative interventions for Alzheimer's disease. Familiar activities, videos can help patients cope with memory loss. PMID- 11899551 TI - Creating a video biography for a loved one who has memory impairment. PMID- 11899552 TI - Vital signs are still ... well, vital. How 'Dr. Rectal,' a stickler for proper technique, never misses a fever. PMID- 11899553 TI - Cognitive and behavioral distancing from the poor. AB - The author argues that distancing is the dominant response to poor people on the part of those who are not poor and that distancing, separation, exclusion, and devaluing operationally define discrimination. Such responses, together with stereotypes and prejudice, define classism. The article focuses on classism in the United States. Classism is examined in the context of theoretical propositions about the moral exclusion of stigmatized others and is illustrated by cognitive distancing, institutional distancing (in education, housing, health care, legal assistance, politics, and public policy), and interpersonal distancing. The adoption of the Resolution on Poverty and Socioeconomic Status by the American Psychological Association Council of Representatives in August 2000 is cited as an important step in the direction of eliminating the invisibility of low-income persons in psychological research and theory. PMID- 11899554 TI - School readiness. Integrating cognition and emotion in a neurobiological conceptualization of children's functioning at school entry. AB - The author examines the construct of emotionality, developmental relations between cognition and emotion, and neural plasticity and frontal cortical functioning and proposes a developmental neurobiological model of children's school readiness. Direct links are proposed among emotionality, use-dependent synaptic stabilization related to the prefrontal cortex, the development of executive function abilities, and academic and social competence in school settings. The author considers research on the efficacy of preschool compensatory education in promoting school readiness and recommends that programs expand to include curricula directly addressing social and emotional competence. Research should focus on the ontogeny of self-regulation and successful adaptation to the socially defined role of student, the development of prevention research programs to reflect this orientation, and interdisciplinary collaborations that integrate scientific methods and questions in the pursuit of comprehensive knowledge of human developmental processes. PMID- 11899560 TI - Comparison of medical and psychological tests. PMID- 11899559 TI - Validity and values: monetary and otherwise. PMID- 11899561 TI - How should psychological assessment be considered? PMID- 11899562 TI - Psychological testing and psychological assessment: a closer examination. PMID- 11899563 TI - Amplifying issues related to psychological testing and assessment. PMID- 11899564 TI - Scientific freedom is not the only issue. PMID- 11899565 TI - Learning considered within a cultural context. Confucian and Socratic approaches. AB - A Confucian-Socratic framework is used to analyze culture's influence on academic learning. Socrates, a Western exemplar, valued private and public questioning of widely accepted knowledge and expected students to evaluate others' beliefs and to generate and express their own hypotheses. Confucius, an Eastern exemplar, valued effortful, respectful, and pragmatic acquisition of essential knowledge as well as behavioral reform. Expressions of these approaches in modern postsecondary contexts are discussed, as are the effects these approaches may have for students who either fit or do not fit the cultural ideal. PMID- 11899566 TI - [Coronary angioplasty in elderly patients]. AB - Developed nations are experiencing an unprecedented growth in the number of elderly citizens. Thanks to the modernization of both society and medical care, over the past century, life expectancy in most countries has nearly doubled. The elderly (80 years or over) represent the fastest-growing segment of our population. Owing to their age, the elderly are more afflicted with chronic diseases, including coronary artery disease. Recently, clinicians became more aggressive in the use of invasive cardiovascular diagnostic tests in these patients. With regard to younger patients, we have a wealth of data from large, randomized trials that defined which subsets of patients benefit from revascularization therapies. However, because the very elderly were severely underrepresented in these randomized studies, we have almost no information concerning the choice of treatment in these patients. Initial results of percutaneous revascularization procedures in elderly patients come from retrospective analysis performed during the pre-stent era. In these studies a procedural mortality risk 5-fold higher in patients > 80 years compared with those < 60 is reported. With the advent of coronary stenting, a significant increase in the rate of procedural success and a reduction in the incidence of procedural mortality, acute myocardial infarction and emergency coronary artery bypass grafting were observed also in the elderly. These initial positive results prompted physicians to treat patients with impaired clinical conditions and with unfavorable angiographic characteristics, resulting in a long-term freedom from major adverse cardiac events comparable to those observed in younger patients (78% in patients < 70 years vs 75% in patients > 80 years). The comparison between the percutaneous approach and the surgical approach to coronary artery disease in the elderly has its major limitation in the lack of data from randomized trials. The available information suggests similar results with the two different strategies (5-year survival rate: 85.7% for the coronary artery bypass grafting group vs 81.4% for the percutaneous revascularization group) with a clear benefit in favor of surgery only in diabetic patients. The only constant difference is the need of a repeat intervention which is significantly higher for the percutaneous approach. Furthermore, the introduction of newer percutaneous devices suitable for the recanalization of totally occluded coronary arteries could increase the completeness of the revascularization achieved with the percutaneous approach with a positive effect on the long-term outcome. These considerations could be "out of time" if the results obtained with the use of drug-eluting stents observed in selected patient populations will be confirmed in larger trials and in routine clinical practice. Whereas the advantages of the percutaneous versus the surgical approach in the elderly have not been fully clarified in patients with stable or unstable angina, primary angioplasty appears to be a very promising strategy in old patients with acute myocardial infarction. The improved clinical success with percutaneous revascularization in the elderly (acute mortality in patients > 85 years: 28.4% reperfused vs 38.5% not reperfused; p = 0.001) has to be related to the use of stents and to IIb/IIIa antagonists. In conclusion, when an old patient needs myocardial revascularization, the percutaneous approach should, in our opinion, be considered the treatment of choice in subjects either with stable or unstable angina and in those with acute myocardial infarction. At present, surgical revascularization provides better results in diabetic patients. The advent of drug-eluting stents may change many of the current limitations of percutaneous revascularization and further expand the use of this strategy. PMID- 11899567 TI - [Invitation to "slow medicine"]. AB - In clinical practice, hyperactivity is often unnecessary. Adopting a strategy of "slow medicine" may be more rewarding in many situations. Such an approach would allow health professionals and particularly doctors and nurses, to have a sufficiently long time to evaluate the personal, familial and social problems of patients extensively, to reduce anxiety whilst waiting for non urgent diagnostic and therapeutic procedures, to evaluate new methods and technologies carefully, to prevent premature dismissals from hospital and finally to offer an adequate emotional support to the terminal patients and their families. PMID- 11899568 TI - [Transesophageal echocardiography diagnosis of ruptured aneurysm of the Valsalva sinus associated with aneurysm of the interatrial septum]. AB - Aneurysms of the sinus of Valsalva are uncommon heart defects that often remain undetected unless rupture occurs. They have been reported in association with other cardiac anomalies. The present case report deals with a 51-year-old man who was referred to our division with a diagnosis of recent-onset progressive heart failure. Echocardiographic evaluation, both transthoracic and transesophageal, disclosed rupture of an aneurysm of the non-coronary sinus of Valsalva into the right atrium. This anomaly was associated with an aneurysm of the atrial septum. PMID- 11899570 TI - [Unusual case of interventricular septal defect in tertiary syphilis]. PMID- 11899571 TI - [Coronary angioplasty in subgroups at risk: patients treated with bypass]. AB - Percutaneous coronary intervention represents an established method to obtain revascularization in patients suffering from obstructive coronary artery disease. Despite the predictability of procedural results and the favorable clinical outcome shown in large series, there are still areas in which the clinical benefits of intervention are less impressive and a lot remains to be done to improve both procedural and clinical outcomes. Among these areas, one may well include the subgroup of patients with previous bypass surgery, who have been consistently shown to be affected by a high rate of periprocedural complications as well as late recurrence. The risk profile of these patients is examined under two main perspectives: the burden of more severe baseline clinical conditions (age, ventricular function, severity of coronary disease) and the negative impact of graft atherosclerosis. The basic assumption of this article is that a variable combination of these characteristics identifies subsets with increasing risk of complications and/or recurrence. For this reason, results of percutaneous revascularization in these patients may still represent a technical as well as a clinical challenge. In particular, the long debated relationship between composition of atherosclerotic plaque in saphenous vein graft, distal embolization, periprocedural myocardial damage and early and late adverse events represents a negative sequence that currently available pharmacologic and interventional resources cannot consistently antagonize. Added to this, are the unresolved issues of diffuse degeneration and chronic total occlusion of saphenous vein grafts, in which no therapeutic approach, alone or in combination, has substantially modified the poor outcome of these lesions. The use of glycoprotein IIb/IIIa receptor antagonists is also discussed, in the light of available data derived from large clinical trials, casting doubts on the efficacy of these otherwise essential pharmacologic agents. Lastly, the setting of acute myocardial infarction represents the clinical scenario in which the adverse effects of the combination of clinical and angiographic characteristics are clearly appreciated. Both coronary intervention and cardiac surgery represent fields of rapidly growing knowledge and technology. It is likely that in the near future we will witness major changes in the clinical management of these patients, thanks to the increasing utilization of arterial conduits, the widespread use of local drug delivery, the availability of new percutaneous devices, and the development of integrated pharmacologic and mechanical revascularization strategies. PMID- 11899572 TI - [Echocardiography in patients with acute cerebrovascular syndromes. Rational use and clinical impact]. AB - About 30% of acute cerebrovascular attacks have a cardioembolic etiology. It is well known that intracardiac thrombi or tumors are related to acute cerebrovascular attacks. However, the role of other cardiac lesions is not so clear. Echocardiography, in particular with transesophageal echocardiography, has the potentiality to correctly identify the cardiac source. In the clinical scenario, the diagnosis is important but the clinical impact in terms of therapy should also be considered. The aim of this paper was to review the potential role of echocardiography in patients with an acute cerebrovascular attack and to propose an evaluation flow-chart of these patients. PMID- 11899569 TI - [Rheolytic percutaneous thrombectomy with Angiojet (Possis) catheter in acute coronary syndromes. Report of 3 cases and technical considerations]. AB - Mechanical thrombolysis in acute coronary syndromes is poorly understood and is still considerably underused in interventional cardiology. The authors report 3 cases of thrombotic coronary obstruction successfully treated with the Possis Angiojet system. In the first case, coronary angiography which demonstrated a large subocclusive thrombus in the mid right coronary artery was performed in a patient with unstable angina following acute myocardial infarction. Removal of the bulk of the thrombus through the activation of the Angiojet system allowed safe and direct stenting of the residual obstruction. In the second case, the Angiojet system was used to remove a thrombus which prolapsed within a stent during a primary angioplasty procedure on the left anterior descending coronary artery. The third case demonstrates the efficacy of the Possis Angiojet system in cleaning up an extensively thrombosed vein graft of an elderly patient with acute anterior myocardial infarction and cardiogenic shock. After thrombus debulking and restoration of TIMI 3 flow in the graft and in the native left anterior descending coronary artery, the left ventricular function was restored and the hemodynamic picture improved. This technique, the indications and results of this promising new device are discussed and reviewed in detail. PMID- 11899573 TI - [In patients with heart failure, do implantable cardioverter-defibrillators delay death or save lives?]. AB - Heart failure is one of the most important public health problems in western countries because of its frequent association with cardiac death and with rehospitalization. Patients with heart failure generally die of sudden arrhythmic death (SD) and progressive pump failure with a SD incidence inversely related to the severity of the underlying heart disease. SD occurs approximately in one half of the patients in New York Heart Association (NYHA) functional classes II-II/III and in one third of those in NYHA classes III/IV-IV, respectively. In the last decade, numerous studies have shown that the automatic implantable cardioverter defibrillator (ICD) significantly reduces the incidence of SD in those patients who are identified as being at risk. Unfortunately, little is known on whether the ICD actually prolongs the survival of the subgroup of patients with most severe heart failure. The main reasons for such a paucity of information are the small number of available studies, the unavailability of randomized and controlled investigations and the difficulty in comparing the results of such studies owing to the lack of homogeneity. In patients with severe heart failure, both the perioperative mortality and morbidity related to transvenous ICD implantation are similar to those of patient subgroups with moderate or slight heart failure. The defibrillation threshold at implantation and the frequency of intractable ventricular arrhythmias during follow-up (2% of all ICD implantations) are slightly higher than in the patient subgroup with moderate or no heart failure. In the patient subgroup with severe heart failure or with a very poor left ventricular function, ICD implantation is unable to prolong the 1- and 2-year survival despite a clear reduction in the incidence of SD. On the other hand, in the patient subgroup with moderate left ventricular dysfunction, ICD implantation prolongs survival and reduces the incidence of SD. No information is available regarding the primary prevention of SD in patients with heart failure. PMID- 11899574 TI - [Patterns of information and informed consent procedures. Study of the Ethical Commission of the ANMCO]. AB - A survey on patterns of information and informed consent procedures during daily clinical practice has been performed by the Ethical Commission of ANMCO. A structured questionnaire (38 questions) was sent to the 653 cardiological units of the National Health Service in Italy. Four hundred and eighty (73.5%) were received. The following variables were considered to evaluate differences in the answers from the various cardiological units: geographical site, presence or absence of an in-patient department, a cath-lab, and of cardiac surgery facilities. Independent predictors of returning questionnaires were: geographical site (Northern Italy vs Central and Southern) and the presence of a cath-lab. Informed consent forms were provided in 53% of instances, while in 40% a free comment about the topic of informed consent was sent. Statistically significant differences in the answers were found about physicians' and nurses' role, ways of information, qualitative and quantitative risk estimates, other persons' role, models of consent forms and procedures of obtaining consent. Free comments and informed consent forms did not allow a statistical analysis. However, they still provided sufficient material to identify specific patterns of how cardiologists deal with the informed consent process. The distinction between the two phases of information and consent was rarely clear. Information or educational material was often mixed with consent forms. While some still showed a paternalistic approach, or else considered informed consent as a formal act, others demonstrated a deep understanding of the significance of the concept of informed consent. A widespread need of guidelines and standard patterns resulted. PMID- 11899575 TI - [Hospital statistics as tool in epidemiologic studies: heart failure in Trieste]. AB - BACKGROUND: Since 1995, the reimbursement of hospital healthcare expenditure in Italy has been based upon the so-called Diagnosis Related Groups (DRGs). The DRG 127 includes all the cases in which the main clinical diagnosis is "heart failure or shock" and, therefore, it may be used to obtain epidemiological data concerning this syndrome. The analysis appears to be of relevance, since in the district of Trieste the phenomenon of progressive aging of the general population has already reached a very advanced phase. METHODS: In this study, we evaluated, using the database of the Sistema Informativo Sanitario Regionale, the epidemiological data and clinical outcome of patients hospitalized for DRG 127 in the district of Trieste from 1997 to 2000 (5514 hospital admissions and 69,236 days of hospital stay). RESULTS: The DRG 127 accounted for 2.6% of the total hospital admissions and 4% of the total days of hospital stay; moreover, it was found to be the first cause of hospitalization for medical DRGs (18.8%) as well as the first cause of hospitalization and of the days of hospital stay for cardiovascular DRGs (27.5 and 40.5%, respectively). Seventy-two percent of patients admitted for DRG 127 were > 75 years. In 1997, a mean of 4/1000 inhabitants of the district of Trieste were hospitalized for DRG 127 (4.6 hospital admissions and 63 days of hospital stay/1000 inhabitants). Over the 4 year period, the number of hospital admissions for DRG 127 increased by 20.4%, the days of hospital stay by 11.1%, and the related healthcare costs rose by 36.7%. Most of the patients (89%) were admitted in Internal Medicine or Geriatric wards. By using suitable corrective factors, the prevalence rate of heart failure in 1997 was estimated at 6.4@1000 (< 65 years 1.8@1000, 65-74 years 10.3@1000, 75 84 years 22.3@1000, > or = 85 years 47.4@1000). On the basis of the first hospital admission, the incidence was estimated at 2.4@1000 (< 65 years 0.8@1000, 65-74 years 2.2@1000, 75-84 years 9.3@1000, > or = 85 years 20.4@1000). The in hospital 1-, 2- and 3-year survival rates from the time of the first hospital admission for heart failure were, respectively, 92, 80, 68 e 56%. The most frequent etiology was ischemic heart disease (50%), followed by hypertensive (26%) and valvular (7%) heart diseases. Ischemic patients showed the worst prognosis and ischemic heart disease was associated with a 3-year survival rate of 44%. However, the prognosis was found to be significantly influenced by the age of the patients, independently of the disease etiology. CONCLUSIONS: The analysis of the hospitalizations for DRG 127 in the district of Trieste may contribute to forecast the epidemiological scenario and the healthcare demand for heart failure that will occur in the next decades at a national level, and may be useful to plan effective models of integrated home-hospital care for the increasing number of affected subjects. PMID- 11899576 TI - [Coronary angioplasty in women: risk factors and sex-related differences in coronary anatomy evaluated with intravascular ultrasonography]. AB - BACKGROUND: The aim of this study was to evaluate the risk factors for cardiovascular diseases, clinical presentation and coronary anatomical size differences in women. METHODS: From January 1999 to December 2000, 244 female and 980 male patients were submitted to coronary angioplasty (PTCA). For both groups the following were considered: risk factors for cardiovascular diseases, clinical presentation and angiographic data. The clinically confirmed 6 months restenoses were evaluated. We performed intravascular ultrasound (IVUS) with three dimensional reconstruction and quantitative coronary angiography (QCA) on the proximal left anterior descending (LAD) coronary artery segments free of significant atherosclerosis in 60 men and 50 women matched for age and clinical characteristics. The arterial and luminal areas were measured by planimetry and corrected for body surface area. We also evaluated the external elastic membrane diameter (EEMd), the minimal lumen diameter (MLD) and the intima-media thickness (IMT). RESULTS: At the time of admission, women were older than men, were shorter, weighed less, and had a smaller body surface area; they had more severe angina, diabetes mellitus and hypercholesterolemia. There was no difference between women and men in the incidence of clinical restenosis at 6 months of follow-up. At IVUS, the mean uncorrected LAD arterial area was smaller in women than in men (12.7 +/- 3 vs 15.9 +/- 3.3 mm2, p < 0.05), as was the mean LAD luminal area (9.9 +/- 3 vs 12.9 +/- 2.7 mm2, p < 0.005). Both the MLD and the EEMd as well as the IMT were smaller in women than in men (MLD 3.3 +/- 0.6 vs 3.9 +/- 0.5 mm, p < 0.05; EEMd 3.7 +/- 0.6 vs 4.2 +/- 0.4 mm, p < 0.005; IMT 0.29 +/- 0.1 vs 0.4 +/- 0.1 mm, p < 0.05). QCA confirmed the IVUS results (MLD 2.9 +/- 0.6 vs 3.5 +/- 0.8 mm, p < 0.05). After correction for body surface area, univariate associations between sex and both the arterial and luminal areas were no longer present. CONCLUSIONS: Women submitted to PTCA were older. The incidence of hypertension, diabetes mellitus and hypercholesterolemia was higher than in men. There was no sex difference in the rate of clinical restenosis at 6 months of follow-up. The LAD artery is smaller in women, independently of body size. This suggests an intrinsic sex effect on coronary dimensions. PMID- 11899577 TI - [Atrial fibrillation: always cardioversion? Yes]. AB - It is obvious that sinus rhythm is preferable to atrial fibrillation for the improvement of the clinical conditions of patients and to avoid the risks of anticoagulation. This clinical reality associated with recent scientific evidence, i.e. atrial fibrillation can be treated with focal ablation, a fibrillating atrium undergoes an unfavorable remodeling that can negatively influence attempts of electrical cardioversion, results of clinical studies have demonstrated that a large atrial size and long-standing atrial fibrillation do not represent a contraindication to electrical cardioversion, explain why at present cardiologists try to maintain normal sinus rhythm more than in the past. Results of current studies that compare rate control versus sinus rhythm maintenance could solve the controversy. PMID- 11899578 TI - [Atrial fibrillation: always cardioversion? No]. AB - Two major treatment strategies have emerged in the management of patients with atrial fibrillation (AF): restoration of sinus rhythm and antiarrhythmic drug prophylaxis versus ventricular rate control and chronic anticoagulation. Besides the potential benefits of the restoration of sinus rhythm, several considerations support the choice of controlling the heart rate, mainly the poor efficacy of antiarrhythmic drug prophylaxis. The decision of pursuing the AF cardioversion should be based mainly on the importance of sinus rhythm restoration and the probability of sinus rhythm maintenance. The factors conditioning the maintenance of sinus rhythm following cardioversion are the duration of AF, cardiac size and function, underlying heart disease, the NYHA functional class, and the timing and number of AF recurrences. At least one attempt at cardioversion is warranted in the majority of patients with a first ever episode of AF; however, it seems advisable to give up even the first attempt at cardioversion in the mildly symptomatic patients who are very old, in patients with AF episodes dating back more than 24-36 months and in those with severe valvular heart disease or severe left ventricular dysfunction. A repeated attempt at cardioversion is usually indicated at the first recurrence of AF; repeated cardioversion seems unadvisable in patients with long-standing AF and early recurrence, in case of failure of amiodarone prophylaxis or of side effects of antiarrhythmic drugs, and when the patient is inclined not to undergo a new electrical cardioversion procedure. In patients with further recurrences of AF it is convenient to give up the cardioversion in case of mild symptoms, of failure of several antiarrhythmic drug regimens and when the withdrawal of oral anticoagulant therapy following sinus rhythm restoration is not safe. With regard to mortality, morbidity, quality of life and cost-effectiveness, the strategy of choice has not yet been established. Several large prospective randomized clinical trials comparing cardioversion and antiarrhythmic prophylaxis versus ventricular rate control are ongoing. The results of these studies could, in the near future, provide useful indications for the choice of the therapeutic regimen to be employed. PMID- 11899579 TI - [Coronary angioplasty in diabetic patients]. AB - Increased thrombogenicity, endothelial dysfunction, proliferation of both the cells and the matrix in the vessel walls, dislipidemia and insulin-resistance are the main metabolic alterations in the diabetic scenario, with possible implications in terms of vessel restenosis after coronary angioplasty. The outcome of balloon coronary angioplasty in diabetics is poor. This is due to both increased restenosis and a high incidence of medium/long-term cardiac events; the use of stents in these patients has substantially improved the results, but the recurrence rate has not been abated to the level observed in the general population. Abciximab may be a helpful adjunct to coronary angioplasty in these patients, while coronary artery bypass grafting may still be preferable--as at present--in case of multivessel coronary artery disease. This viewpoint is likely to be substantially modified in the near future, if the promise of "zero restenosis" devices is kept. PMID- 11899580 TI - [From prevention to cardiovascular rehabilitation: statins and evidence-based medicine]. AB - The recent "Lipobay affair" prompted us to review the biomedical literature focusing on statins, in a prevention and rehabilitation perspective, and to prepare a pertinent evidence-based report. A number of randomized controlled clinical trials have assessed the efficacy and the safety of statins in cardiovascular prevention; these drugs have proved to be effective (number needed to treat values between 20 and 40) and safe (number needed to harm value for myopathy even when in association with fibrates around 833, as deduced from literature data). Fatal rhabdomyolysis is virtually not detectable in current scientific literature when statin therapy is administered in an appropriate fashion and when the patient undergoes periodical check-ups. In the individual patient, the benefits and risks related to his treatment have to be precisely tailored, evaluating his individual risk profile, the strength of the indication to statin therapy, the concomitant presence of other treatments and/or habits and the potential and actual adverse events foreseeable (to an extent) if established risk factors which predispose to myopathy are taken into consideration. The hematological lipids management strategy including a correct diet, physical exercise and statin therapy represents a key component of every evidence-based rehabilitation program too. Cardiovascular prevention and rehabilitation are major priorities at an individual as well as at a public health level. Both these strategies are based on a careful management of known risk factors, among them hypercholesterolemia, that today can be effectively and safely treated with statins. PMID- 11899581 TI - [Pulmonary scintigraphy in the diagnosis of pulmonary embolism]. AB - Pulmonary scintigraphy constitutes an important step in the non invasive diagnosis of pulmonary embolism (PE). This technique may be employed for the evaluation of the pulmonary perfusion alone, as in Italy and in the PISA-PED study, or else even for the evaluation of the pulmonary ventilation (as in Anglo Saxon countries and in the PIOPED study). In the present study, the reasons which have prompted the ANMCO-SIC Commission for the Guidelines for The Prophylaxis, Diagnosis and Therapy of Pulmonary Thromboembolism to propose, for the diagnostic work-up of the patient with clinically suspected PE, the use of perfusion scintigraphy alone and of the classification criteria employed in the PISA-PED study instead of the more commonly utilized ventilatory-perfusion scintigraphy and of the criteria included in the PIOPED article, are discussed. Besides, the Commission's decision to consider PE as being present in case of agreement between the scintigraphic and clinical pictures, and to exclude this condition when the scintigraphic outcome is normal/almost normal regardless of the clinical probabilities, is also motivated. PMID- 11899582 TI - Analysis of gene-environment interactions by "stressing-the-genotype" studies: the angiotensin converting enzyme and exercise-induced left ventricular hypertrophy as an example. AB - The human angiotensin converting enzyme (ACE) gene contains a length polymorphism consisting of the presence (insertion, I) or absence (deletion, D) of a 287 base pair "Alu" repeat sequence in intron 16, with the D allele being associated with higher ACE levels than the I allele in plasma and in tissues. We have carried out several studies to examine the relationship between this polymorphism and cardiovascular health, and have examined the hypothesis that if renin-angiotensin systems regulate left ventricular (LV) growth, individuals of DD genotype might show a greater hypertrophic response than those of II genotype. A strategy was used involving screening over 1200 male military recruits to select only subjects homozygous for the I or D allele for the expensive and time-consuming but extremely accurate method of LV mass determination by magnetic resonance imaging. LV dimensions and mass were compared at the start and end of a 10-week physical training period. LV mass increased with training by 8.4 g overall (p < 0.0001), but with DD men showing roughly 3 fold greater growth than II men (p < 0.001). When indexed to lean body mass, LV growth in II subjects was essentially negligible whilst remaining significant in DD subjects (-0.022 vs +0.131 g/kg respectively, p = 0.0009). Although the precise molecular mechanism of this effect remains to be elucidated it clearly demonstrates the importance of the ACE renin-angiotensin system in determining LV dimensions in situations of high cardiac demand, which may also be important in pathology such as hypertension and heart failure. The use of these "stress-the-genotype" approaches to explore gene environment interactions are likely to be the key to understanding the causes determining both coronary artery disease and other multi-factorial disorders. PMID- 11899583 TI - How to search for the role of gene-environment interactions for lipids in humans. AB - To target the interventions to the requirements of the individuals we need to identify the genes and the alleles which show variations in response pattern. For example, a lipid-lowering diet is effective for most people, but not so for everyone. The search for genetic polymorphisms that affect the response of LDL cholesterol to diet in humans has been disappointing up till now, and is the first hurdle to take. In addition, it should be realized that the phenotype of serum lipid is not only indicated by the blood level, but is multifaceted. A complication of gene-environment interactions is that in the chain of environmental factor, intermediate phenotype, and pathogenetic process we have the possibility of genetic variability at all levels. PMID- 11899584 TI - Analysis of gene-environment interaction in coronary heart disease: fibrinogen polymorphisms as an example. AB - Several epidemiological studies have shown that an increase in fibrinogen levels is associated with the risk of cardiovascular disease. Recently, it has been demonstrated that the levels of fibrinogen can be genetically determined. Overall the studies show a strong association between two polymorphisms of the fibrinogen beta-chain gene and fibrinogen plasma concentration. Few studies have, in contrast, found an association between such polymorphisms and the risk of ischemic vascular disease. Rather than directly affecting the levels of proteins or the risk of disease, polymorphisms can amplify the effect of environmental or intermediate conditions on the final phenotype. The genetic control of fibrinogen has to be considered together with environmental factors: fibrinogen genotypes may interact with cigarette smoking, gender, physical activity, use of drugs and infections in determining the increase in fibrinogen levels and perhaps the risk of ischemic heart disease. Three examples are presented supporting the concept that, in multifactorial diseases, genetic variability influences the risk of disease by determining a different individual susceptibility to environmental risk factors. PMID- 11899585 TI - Total correction of tetralogy of Fallot: late clinical follow-up. AB - Forty-five years after the first repair of tetralogy of Fallot we have sufficient data to describe the post-surgical history of these patients in terms of survival, quality of life and delayed complications. The long-term results of surgical repair during infancy and childhood are good in terms of health assessment and exercise capacity. However arrhythmias and right ventricular dysfunction secondary to ventriculotomy and residual pulmonary regurgitation characterize the delayed follow-up. The identification of the clinical parameters which are predictive of premature ventricular dysfunction and electrical instability is a primary aim of clinical follow-up. PMID- 11899587 TI - Analysis of gene-environment interaction in coronary artery disease. PMID- 11899586 TI - Comparative short-term prognostic value of hemostatic and inflammatory markers in patients with non-ST elevation acute coronary syndromes. AB - BACKGROUND: Recent data show that markers of inflammation, endothelial perturbation as well as activation of the coagulation and fibrinolytic systems are altered in unstable angina. The purpose of this study was to compare the 30 day prognostic value of the indexes of inflammation [interleukin-6 (IL-6)], endothelial activation [von Willebrand factor antigen (vWf)], fibrinolysis [plasminogen activator inhibitor-1 (PAI-1)] and coagulation (F1 + 2), in a consecutive series of patients with non-ST elevation acute coronary syndromes. METHODS: Eighty-eight patients consecutively admitted to the coronary care unit because of chest pain occurring within the previous 24 hours were included in the study. Blood was drawn on admission to the coronary care unit and 72 hours thereafter for the assessment of plasma levels of IL-6, vWf, F1 + 2 and PAI-1. Troponin I serum levels were measured 6 to 12 hours after admission. All patients underwent coronary arteriography. RESULTS: Patients were divided into two groups according to their 30-day outcome: 57 patients (group 1) had an uneventful outcome, whereas 31 patients had an adverse clinical event (4 died, 1 had a Q wave myocardial infarction and 26 had refractory angina). The baseline biochemical variables were similar between group 1 and group 2 patients. Seventy two hours following admission, an increase in the serum levels of IL-6 was observed in 71% of group 2 patients and in 28% of group 1 patients (p = 0.0001). The other measured variables showed significant changes at 72 hours versus entry only in group 1 patients, and no significant difference between the two groups. The areas under the ROC curves were higher for IL-6 (0.72) than for the other variables (0.58 for F1 + 2, 0.52 for vWf and 0.54 for PAI-1). In a multivariate model, including clinical, angiographic, and biochemical variables, only the change in IL-6 over 72 hours was significantly associated with a worse 30-day outcome (odds ratio 8.472, 95% confidence interval 1.030-69.671). CONCLUSIONS: This study shows that a mounting inflammatory process, as indicated by increasing levels of IL-6 over the first 72 hours after admission, is the most powerful predictor of the 30-day prognosis in patients with non-ST elevation acute coronary syndromes. PMID- 11899588 TI - Effects of different training protocols on left ventricular myocardial function in competitive athletes: a Doppler tissue imaging study. AB - BACKGROUND: The aim of the study was to analyze the differences in myocardial function in case of left ventricular (LV) hypertrophy induced by either endurance or strength training in top-level athletes. METHODS: Standard Doppler echo and pulsed Doppler tissue imaging (DTI) of the interventricular septum and of the LV inferior wall were performed in 26 competitive endurance athletes (long-distance swimmers) (group A) and in 20 strength-trained athletes (short-distance swimmers) (group B). By means of DTI, the following parameters of myocardial function were assessed: the systolic peak velocities (Sm), the pre-contraction time, the contraction time, the early (Em) and late (Am) diastolic velocities, the Em/Am ratio and the relaxation time. RESULTS: The two groups were comparable for age and sex, but at rest group B showed a higher heart rate, systolic blood pressure and body surface area. The LV mass index and fractional shortening did not significantly differ between the two groups. However, group B showed an increased sum of the wall thicknesses (septum + posterior wall) (p < 0.001) and of the relative wall thickness, while the LV stroke volume and LV end-diastolic diameter (p < 0.001) were greater in group A. All transmitral Doppler indexes were higher in group A, with an increased E/A ratio. DTI analysis showed, in group A, a higher Em and Em/Am ratio as well as a longer relaxation time both at the septal and at the inferior wall levels, with comparable Sm, pre-contraction and contraction times. Distinct multiple linear regression models revealed an independent positive association between the inferior peak Em velocity and the LV end-diastolic diameter (p < 0.001) in group A, and an independent direct correlation of the inferior peak Sm velocity with the sum of the wall thicknesses (p < 0.001) and with the end-systolic stress in group B. CONCLUSIONS: The early diastolic myocardial function is positively influenced by the preload increase in group A, while an increased afterload and LV wall thickness in group B mainly seem to induce an enhancement of the regional myocardial systolic function. PMID- 11899589 TI - Resting echocardiographic assessment of regional wall motion, thickness and reflectivity in chronic ischemic cardiomyopathy: an alternative to the viability test? AB - BACKGROUND: A resting echo showing a regional end-diastolic wall thickness < or = 6 mm with a hyperechoic texture is pathognomonic of scar tissue and of non-viable myocardium. The aim of this study was to assess the prognostic value of the resting echo scar texture in patients with chronic ischemic cardiomyopathy evaluated prior to coronary artery bypass surgery. METHODS: The preoperative clinical and echocardiographic data of 70 patients with a mean ejection fraction of 29.8 +/- 4% scheduled for coronary revascularization were correlated to the cardiac events observed during a mean follow-up of 24 +/- 12 months after surgery. Akinetic segments of the left ventricular wall with a reduced diastolic thickness and increased echoreflectivity were judged scarred. RESULTS: Sixty eight patients were discharged alive from hospital. On the basis of ROC analysis, we identified: group A (27 patients) with > 5 and group B (41 patients) with < or = 5 scarred segments. There were 10 events (3 deaths, 4 heart transplants and 3 refractory heart failures), 8 in group A (29%) and 2 in group B (5%). At multivariate analysis the only independent predictor of the clinical outcome after revascularization was whether the patient was included in group A or B (Wald 6.3, p < 0.012). One year after surgery, the ejection fraction improved only in group B patients (p < 0.03). CONCLUSIONS: The extent of scarred myocardial tissue as assessed at resting echocardiography predicted the benefit of revascularization in patients with chronic ischemic left ventricular dysfunction. This simple and straightforward echo parameter should be taken into consideration when assessing the instrumental value of more technologically demanding and costly viability testing. PMID- 11899590 TI - Cardiac rhabdomyoma in intrauterine life: clinical features and natural history. A case series and review of published reports. AB - BACKGROUND: Fetal cardiac rhabdomyoma is very rare; despite the fact that many cases and series have been reported, the clinical presentation, the natural history and the frequency with which this pathology is associated with tuberous sclerosis complex are not well determined. The aim of this investigation was to study the clinical features and the natural history of cardiac rhabdomyoma when diagnosed during prenatal life. METHODS: Nine cases of cardiac rhabdomyoma detected among 5276 fetal echocardiograms recorded over a 10-year period in a single center were retrospectively reviewed. Medical records and echocardiograms were studied to determine the prenatal and postnatal course and outcome. RESULTS: The incidence of cardiac rhabdomyoma in our center was 0.17%. The gestational age at diagnosis ranged from 27 to 36 weeks. The most common reason for fetal echocardiography was an abnormal obstetric ultrasound scan (6/9 cases). In no case was there a family history of tuberous sclerosis. In one case, the tumor was single whereas in 8 cases multiple tumors were diagnosed. During prenatal life the majority of tumors were clinically silent. One fetus died of hydrops and arrhythmia. Four children presented with arrhythmia postnatally and one required surgery. At a mean follow-up of 47 months, total or partial regression was observed in 7 patients. Seven patients developed postnatal clinical signs of tuberous sclerosis. CONCLUSIONS: Fetal cardiac rhabdomyomas are often benign and have a tendency to regress, but their prognosis is guarded due to very frequent association with arrhythmias and tuberous sclerosis. During prenatal counseling, it is of utmost importance to inform the future parents of the virtually constant perspective of tuberous sclerosis complex. PMID- 11899591 TI - Behavioral, psycho-physiological and salivary cortisol modifications after short term alprazolam treatment in patients with recent myocardial infarction. AB - BACKGROUND: The aim of this study was to determine the behavioral and physiological effects of the central nervous system depressant alprazolam on a group of cardiac patients. METHODS: Immediately after hospital discharge, the Crown and Crisp Experiential Index (CCEI) was administered, the salivary cortisol was detected and a psycho-physiological profile was recorded in 52 subjects who had suffered from myocardial infarction. Half of the subjects represented the experimental group and the remaining 26 individuals acted as a control group not undergoing treatment. The benzodiazepine alprazolam (0.25 mg) was administered twice daily to the treated group only. With the exception of the administration of the drug, all recruited subjects underwent the same clinical evaluation. RESULTS: The CCEI data of the treated group showed significant decreases for the following scales: free floating anxiety (p < 0.001), phobic anxiety (p < 0.01), somatic complaints (p < 0.05), and depression (p < 0.01). In the same group, with regard to the physiological parameters, the skin conductance response significantly decreased during the baseline phase (p < 0.01), and almost all parameters showed decreased values during mental stress test administration. Cortisol levels also decreased during the recovery phase of the psycho physiological profile assessment. CONCLUSIONS: Alprazolam seems to be able to reduce sympathetic discharge and some stress-related behavioral and physiological responses. This could be of benefit for selected cardiac patients for whom increases in sympathetic tone may constitute a risk factor. PMID- 11899593 TI - Awareness of hypertension guidelines in general practice: a pilot study in Lombardy. AB - BACKGROUND: An adequate knowledge of hypertension guidelines by primary care physicians is a fundamental step for the improvement of the diagnosis and treatment of hypertension in the general population. The aim of this study was to evaluate, in a local survey in the northern area of Lombardy, the general practitioners' knowledge of the WHO/ISH guidelines. METHODS: A 10-item mail questionnaire based on the 1999 WHO/ISH hypertension guidelines was sent to a sample of 280 primary care physicians. The number of answers in agreement with the guidelines was used as a measure of guidelines knowledge, that was considered adequate if a correct answer to 6 out of 10 questions, in addition to an adequate definition of hypertension, was provided. RESULTS: The analysis was based on 83 returned questionnaires, that means a 29% response rate. Guidelines knowledge was adequate in 23.5% of the total study population and the mean score of correct answers was 5.5 points. A significant negative correlation (r = 0.27, p < 0.05) was observed between the mean score of knowledge and the physician's age. CONCLUSIONS: In a sample of primary care physicians from a northern region of Italy, the hypertension guidelines knowledge is inadequate; the reasons and the extent of this poor awareness will require further studies. PMID- 11899592 TI - Analysis of gene-environment interaction in coronary artery disease: lipoprotein lipase and smoking as examples. AB - In a complex disorder such as coronary artery disease (CAD), both genetic and environmental factors influence the onset of disease. Such interactions imply that at the molecular level there is interplay between the gene product and the environmental insult, resulting in a greater than additive effect on risk; for example the synergy between variation in the lipoprotein lipase (LPL) gene and smoking on risk. LPL plays a dual role in lipid metabolism both in the hydrolysis of triglyceride-rich lipoproteins and also as a molecular bridge, enhancing the receptor-mediated uptake of lipoproteins both by the liver (anti-atherogenic) but also by receptors on the artery wall (pro-atherogenic). Smoking is associated with a 2-fold increase in CAD risk, and the mechanisms for this include endothelial damage and promotion of lipid oxidation. Results from a prospective study on CAD risk in healthy middle-aged men show that the risk associated with the LPL-D9N variant, which has a modest effect on plasma triglyceride levels, is enhanced up to 10 fold, but only in men who smoke. The proposed mechanism for this LPL:smoking interaction on CAD risk is the subject of this review. PMID- 11899595 TI - Cigarette smoking in young people: what to search for and what action to take. PMID- 11899594 TI - Factors influencing the age at which adolescents start smoking. A comparison between a big and a small city. AB - BACKGROUND: Most cigarette smokers take up their habit during adolescence. The aim of this study was to compare the age at which the students of a metropolitan high school and those of a non-metropolitan high school start smoking. METHODS: Nine hundred and seventy-eight students (55.6% males, mean age 15.8 +/- 1.5 years) in a big city (Naples) and 467 (50.3% males, mean age 16 +/- 1.5 years) in a small town (Capua-CE) filled in a questionnaire on cigarette smoking. Two hundred and nine (21.4%) students (99 males, 110 females, mean age 16.5 +/- 1.3 years) in Naples and 99 (21.2%) students (59 males, 40 females, mean age 16.8 +/- 1.3 years) in Capua stated that they had smoked at least one cigarette in the last week and were considered as smokers. RESULTS: The age at which adolescents start smoking did not differ between the big and the small city (Naples 14.9 +/- 1.5 years; Capua 14.9 +/- 1.6 years; p = 0.849) nor between sexes, both in Naples (males 14.9 +/- 1.5 years, females 14.8 +/- 1.4 years; p = 0.576) and in Capua (males 14.8 +/- 1.6 years, females 15 +/- 1.5 years; p = 0.379). Both in Naples and in Capua, no relation was found between the age at which the adolescent starts smoking and the smoking habits of the father, mother, siblings, best friend of the same sex, best friend of the opposite sex and friends. In Naples, the age at which the adolescent started smoking was related to the number of cigarettes he or she smoked in the last week (p = 0.004) and to the number of cigarettes smoked per day by the father (p = 0.001). CONCLUSIONS: In adolescents, the age at which the habit of smoking is taken up does not differ both between a big and a small city and between sexes; in the big city, the age at which the adolescent started smoking was related to the number of cigarettes he or she smoked in the last week and to the number of cigarettes smoked per day by the father. PMID- 11899597 TI - Mapping the way to HIPAA compliance. PMID- 11899596 TI - Emergency stenting of the unprotected left main coronary artery. AB - We report a case of successful stenting of the unprotected left main coronary artery as a salvage procedure in a patient with tight ostial left main coronary artery stenosis who had cardiac arrest following diagnostic coronary angiography. PMID- 11899598 TI - Thinking, feeling, doing: keys to learning. PMID- 11899599 TI - Keeping pain control out of the courts. PMID- 11899600 TI - Promoting urinary health. PMID- 11899601 TI - One step purification of corilagin and ellagic acid from Phyllanthus urinaria using high-speed countercurrent chromatography. AB - High-speed countercurrent chromatography (HSCCC) has been successfully applied to the preparative separation of corilagin and ellagic acid in one step from the Chinese medicinal plant Phyllanthus urinaria L. by use of direct and successive injections of a crude methanolic extract. Some aspects concerning the practical use of this technique in the described application are considered. PMID- 11899602 TI - MS/MS profiling of taxoids from the needles of Taxus wallichiana. AB - Ammonium cationisation has been used for taxoid profiling of partially purified methanolic extracts of needles of Taxus wallichiana growing in different regions of the Himalayas (Kashmir, Himachal Pradesh, UP Hills, Darjeeling, Sikkim and Arunachal Pradesh) by electrospray ionisation tandem mass spectrometry (MS/MS). The MS/MS spectra of the [M + NH4]+ or [M + H]+ ions gave structurally diagnostic fragment ions which revealed information about the taxane skeleton as well as the number and nature of the substituents. The rearranged 11(15-->1)-abeo-taxanes showed a characteristic elimination of the hydroxyisopropyl group with an acetoxy/benzoyloxy group from C-9. The identification of the taxoids was achieved by comparison of the MS/MS spectra with those of authentic taxoids or was based on biogenetic grounds. The results were corroborated by liquid chromatography-MS analysis. Out of the 50 taxoids identified, 21 belonged to the rearranged class. The presence of paclitaxel in the samples from four regions was confirmed: the study also revealed the occurrence of several basic taxoids in these samples. MS/MS profiling by electrospray ionisation was shown to be a fast and reliable technique for the analysis of taxoid samples. PMID- 11899603 TI - Liquid chromatography-atmospheric pressure chemical ionisation/mass spectrometry: a rapid and selective method for the quantitative determination of ginkgolides and bilobalide in ginkgo leaf extracts and phytopharmaceuticals. AB - In order to evaluate the composition of active constituents in phytopharmaceutical preparations, valid analytical methods are required. For the determination of the active terpene constituents of Ginkgo biloba (the ginkgolides and bilobalide), a liquid chromatography-mass spectrometry (LC-MS) method has been developed using atmospheric pressure chemical ionisation (APCI) in the negative ion mode. This detection mode was found to be much more sensitive and selective compared to UV; indeed the ginkgo terpene trilactones lack strong UV chromophores and flavonoids interfere with their UV detection. LC-APCI/MS detection allowed a considerable reduction in analysis time when compared to LC UV, because LC resolution was only needed between the pair of isomers ginkgolide B and ginkgolide J. All compounds were selectively detected by single ion monitoring of their specific deprotonated molecules [M-H]-. The samples were directly injected without pre-purification, and a fast gradient was applied, reducing the total time of analysis to 14 min. With this method, the ginkgo terpene trilactones were detected on-line in the picogram range. Several commercial ginkgo preparations on the Swiss market were analysed, and the ginkgolide and bilobalide contents were evaluated using the method described. PMID- 11899604 TI - A rapid capillary gel electrophoresis method for the quantitative determination of RuBisCo in spinach. AB - A capillary gel electrophoretic (CGE) method for the quantitative analysis of RuBisCo in spinach leaves was developed. RuBisCo was resolved into large and small subunits in the presence of sodium dodecyl sulphate (SDS) by the CGE procedure which enabled the determination of the molecular weight of each unit accurately; the values so determined were in close agreement with those reported using other methods. Advantages of CGE over SDS-polyacrylamide gel electrophoresis and high-pressure gel filtration include decreased sample preparation and analysis time, superior resolution and greater sensitivity permitting reduced sample size and trace analysis. In addition, CGE provided precise quantification of RuBisCo and was demonstrated to be a viable alternative to other available methods of protein analysis. PMID- 11899605 TI - Essential oils from micropropagated plants of Lavandula viridis. AB - The essential oils of Lavandula viridis were analysed by GC and GC-MS. Comparisons were made between three types of plant material from the same clone: field-grown plant, in vitro shoot cultures and micropropagated plants of the same clone. The most common components usually found in lavender oils were present in the oil samples analysed and more than 45 constituents were identified, representing more than 80% of the essential oil. The essential oils analysed consisted mainly of monoterpenes (75.4-76.3%), where oxygenated and hydrocarbons identified ranged from 41.8 to 57.3% and 18.1 to 34.2%, respectively. The major components found were 1.8-cineole (18.2-25.1%), camphor (9.1-15.7%), alpha-pinene (8.8-14.1%), borneol (4.1-4.8%), beta-pinene (1.2-5.6%), delta 3-carene (1.0 6.5%) and alpha-terpineol (0.8-4.2%). The monoterpene fraction of the in vitro shoot cultures showed different relative amounts of hydrocarbons and oxygenated components in relation to the parent plant and to micropropagated plants. In the sesquiterpene hydrocarbon fraction of the oil samples analysed (6.1-8.2%), 7-epi alpha-selinene (1.6-4.8%) was the most important component and the oxygenated sesquiterpenes were found in small amounts (1.1-1.7%). The essential oils from field-grown plants of L. viridis, when compared with those obtained from in vitro shoot cultures or micropropagated plants of the same clone, demonstrated that the same major components were found without significant compositional variations. PMID- 11899606 TI - Method for the extraction of the volatile compound salicylic acid from tobacco leaf material. AB - Salicylic acid (SA) is a signalling compound in plants which is able to induce systemic acquired resistance. In the analysis of SA in plant tissues, the extraction recovery is often very low and variable. This is mainly caused by sublimation of SA, especially during evaporation of organic solvents. Techniques have been designed in order to overcome this problem. In the first part of the extraction procedure, sublimation of SA was prevented by addition of 0.2 M sodium hydroxide. At a later stage of the extraction procedure, sublimation of SA during solvent evaporation was controlled by the addition of a small amount of HPLC eluent. In this way, recoveries in the range of 71-91% for free SA and 65-79% for acid-hydrolysed SA were obtained. Recoveries could be further optimised by the use of an internal standard to correct for volume changes after the addition of the HPLC eluent. PMID- 11899607 TI - A rapid TLC bioautographic method for the detection of acetylcholinesterase and butyrylcholinesterase inhibitors in plants. AB - A simple and rapid bioautographic enzyme assay on TLC plates has been developed for the screening of acetylcholinesterase and butyrylcholinesterase inhibition by plant extracts. Enzyme activity was detected by the conversion of naphthyl acetate into naphthol and the formation of the corresponding purple-coloured diazonium dye with Fast Blue B salt. Inhibitors of cholinesterases produced white spots on the dye-coloured background of the TLC plates. The alkaloids galanthamine and physostigmine, which are known inhibitors of acetylcholinesterase, were used to determine the sensitivity of the assay. Various plant extracts were tested using the bioassay. PMID- 11899608 TI - Current awareness in phytochemical analysis. PMID- 11899609 TI - Screening of plant extracts for antioxidant activity: a comparative study on three testing methods. AB - Three methods widely employed in the evaluation of antioxidant activity, namely 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, static headspace gas chromatography (HS-GC) and beta-carotene bleaching test (BCBT), have been compared with regard to their application in the screening of plant extracts. The strengths and limitations of each method have been illustrated by testing a number of extracts, of differing polarity, from plants of the genus Sideritis, and two known antioxidants (butylated hydroxytoluene and rosmarinic acid). The sample polarity was important for the exhibited activity in the BCBT and HS-GC methods but not for the DPPH method. The complex composition of the extracts and partition phenomena affected their activity in each assay. The value of the BCBT method appears to be limited to less polar samples. Although slow, the HS-GC method is preferable for assessing the antioxidant inhibitory properties on the formation of unwanted secondary volatile products. Being rapid, simple and independent of sample polarity, the DPPH method is very convenient for the quick screening of many samples for radical scavenging activity. PMID- 11899610 TI - Web alert. The chemistry of process development. PMID- 11899611 TI - High-throughput techniques for compound characterization and purification. AB - A new paradigm in drug discovery is the synthesis of structurally diverse collections of compounds, so-called libraries, followed by high-throughput biological screening. High-throughput characterization and purification techniques are required to provide high-quality compounds and reliable biological data, which has led to the development of faster methods, system automation and parallel approaches. This review summarizes recent advances in support of analytical characterization and preparative purification technologies. Notably, mass spectrometry (MS) and supercritical fluid chromatography (SFC) are among the areas where new developments have had a major impact on defining these high throughput applications. PMID- 11899612 TI - Advances in high-throughput mass spectrometry. AB - The evolution of high-throughput drug discovery is readily apparent as the pharmaceutical industry continues to stress the rapid progression of new chemical entities and biological agents through drug discovery and development pipelines. Mass spectrometry and high performance liquid chromatography-mass spectrometry have played an instrumental role in the support and advancement of all facets of high-throughput drug discovery. The introduction of new instrumentation has extended the breadth of mass spectrometric-based capabilities from the characterization of high-throughput organic synthesis products to early adsorption, distribution, metabolism and excretion profiling. Additionally, advances in the capacity and throughput of mass spectrometry systems have concurrently led to the introduction of data management tools to address automated data reduction, archival and mining, as well as analytical data integration to chemical and biological databases. PMID- 11899613 TI - Polymer supports in organic catalysis and synthesis. AB - Recent developments in the preparation of polymer supports for synthesis and catalysis are reviewed. The use of insoluble supports for synthesis of one molecule per bead and chemistry leading to new ways to use insoluble supports are discussed. New routes to design soluble, high capacity supports for synthesis are highlighted and the roles of polymers in catalysis, both for re-use and separation, are discussed using metathesis catalysts and Pd(0) catalysts as examples. The role of dendrimers as supports for synthesis and catalysis are reviewed, and new separation schemes using inverse temperature-dependent solubility of polymers and liquid/liquid separations are discussed. PMID- 11899614 TI - Developments in hydrogenation technology for fine-chemical and pharmaceutical applications. AB - The continuous innovation in hydrogenation technology is testimony to its growing importance in the manufacture of specialty and fine chemicals. New developments in equipment, process intensification and catalysis represent major themes that have undergone recent advances. Developments in chiral catalysis, methods to support and fix homogeneous catalysts, novel reactor and mixing technology, high throughput screening, supercritical processing, spectroscopic and electrochemical online process monitoring, monolithic and structured catalysts, and sonochemical activation methods illustrate the scope and breadth of evolving technology applied to hydrogenation. PMID- 11899615 TI - Development of fermentation process for rDNA products. AB - Fermentation process development for recombinant DNA-derived products is becoming increasingly important in the current commercial and regulatory framework. This article provides an overview of the current approach to process development, and the contribution of developmental data to final process validation is highlighted. Cited literature is restricted to between 1995 and 2001. PMID- 11899616 TI - Alternative methods of terminal sterilization for biologically active macromolecules. AB - The traditional perception within the pharmaceutical industry of the manufacture of injectable drug products is that active pharmaceutical ingredients (API) that are peptides, proteins or biopolymers, such as poly(DL-lactide) (PLA) and poly(DL lactideco-glycolide) (PLGA), cannot be terminally sterilized. This perception exists largely because terminal sterilization is assumed by many to be only carried out by steam sterilization in a standard autoclave. Thus, it is understood that these API candidates must be manufactured by aseptic techniques. With the current technological advances in the area of protein and peptide sterilization, which has largely come from the food industry and has in recent years been developed for pharmaceutical use, techniques have been developed for the terminal sterilization of thermally sensitive APIs and biopolymers. In this review, the focus will be on the four major types of sterilization that are presented in the literature: (i) gamma-irradiation; (ii) e-Beam; (iii) natural light; and (iv) microwave. Each of these sterilization techniques present advantages and disadvantages for use in large-scale terminal sterilization of bioactive macromolecules. PMID- 11899617 TI - Spectroscopic methods for determining enantiomeric purity and absolute configuration in chiral pharmaceutical molecules. AB - Analytical support, such as methods development, along with identification and characterization of intermediates and impurities, are critical in the development of a chemical process. The preparation of a drug substance requires the development of analytical methods for monitoring reactions and identifying impurities. Methods development for a chiral drug molecule is more difficult as the method must be capable of monitoring the overall reaction as well as possible racemization of starting materials and products. Chiral methods are often required to monitor the reaction steps of a synthesis, however, the development of enantiomeric purity methods are time-consuming and expensive. The use of chiroptical detectors, such as circular dichroism (CD), optical rotation (OR) and vibrational circular dichroism (VCD), can help to reduce or eliminate the need to develop chiral monitoring methods and also to predict absolute configuration. Recently, VCD has shown remarkable success with the latter and currently holds the most promise as a general, direct method that can be used as an alternative to X-ray crystallography. Each of the mentioned techniques can help analytical chemists to reduce the time associated with traditional enantiomeric purity methods development and to determine absolute configuration. This review will discuss the scope and limitations of these techniques for the rapid and routine determination of both enantiomeric excess and absolute configuration. PMID- 11899618 TI - Synthesis of influenza neuraminidase inhibitors. AB - Influenza neuraminidase inhibitors provide a means to combat flu, a potentially very serious disease. For the first time, there is a way to treat influenza by blocking the influenza enzyme neuraminidase to stop or slow the progression of infection. The diverse structures and synthesis of several influenza neuraminidase inhibitors are discussed. Contributions from chemical process development groups are highlighted for those compounds that have reached the market, such as zanamivir and oseltamivir phosphate. PMID- 11899619 TI - The selection of a commercial route for the D1 antagonist Sch-39166. AB - D1 Antagonists have been reported to be potentially useful in a number of therapeutic areas. Sch-39166 is an example of such a selective D1 antagonist. Three different routes based on (1S,2S)-1-phenyl-1,3-propanediol (19), (+)-L homophenylalanine (27) or trans-(+)-(1R,2R)-2-hydroxy-1-(methylamino)-1,2,3,4 tetrahydronaphthalene (37) were developed for the commercial preparation of Sch 39166. After analyzing each route for the best combination of cost, yield, throughput and efficiency, the synthesis of Sch-39166 starting from 37, which involves the opening of an aziridinium, was selected for optimization into the commercial route. PMID- 11899620 TI - Methods for the synthesis of unnatural amino acids. AB - A wide variety of methods have been used to prepare unnatural amino acids. This review covers methods reported between 2000 and the early part of 2001. Some of the approaches discussed are applications of established methods, but emphasis has been given to new approaches that could be generally applicable to large scale syntheses, including catalytic reactions such as the Strecker reaction and biological approaches. PMID- 11899621 TI - The practical synthesis of vitamin D analogs: a challenge for process research. AB - New, highly-potent vitamin D analogs have increasingly come under consideration for the treatment of a variety of diseases as diverse as psoriasis, diabetes, renal osteodystrophy, osteoporosis, leukemia, cancer (breast, colon, prostate), AIDS and multiple sclerosis. This review will present recent efforts for the development of practical syntheses of these valuable compounds using the synthetically convergent Lythgoe approach. PMID- 11899623 TI - Oops! He did it again. PMID- 11899622 TI - Safety issues in scale-up of chemical processes. AB - Fast time-to-market is an important issue in the development of chemical processes for fine chemicals and drug production. However, this needs to be balanced against the equally important issue of process safety. Developing a safe process within a short time frame is a demanding challenge but advances in: (i) risk analysis methods; (ii) procurement and interpretation of safety and scale-up data; and (iii) process control have opened up new perspectives in this field. The thermal stability of chemicals during storage and transportation is another field of interest, and data obtained from research in this area should allow the simplification of certain tedious procedures. PMID- 11899624 TI - Utilization of medication-assistance programs for medically uninsured patients: one public teaching hospital's experience. AB - Although many pharmaceutical companies have programs that provide prescription medications to medically uninsured or underinsured patients, many health-care providers have not taken full advantage of the programs for a variety of reasons. In an effort to offset the escalating costs of medications for this group of patients, one public teaching facility developed a program to better utilize pharmaceutical patient-assistance programs. Resources are described, including Web sites and available publications, that may improve utilization of assistance programs. Eligibility criteria and enrollment procedures for specific drug reimbursement programs are included. Enrollment of medically underinsured or uninsured patients into medication-assistance programs can produce significant savings that benefit patients and institutions. Healthcare providers must examine all avenues to provide medications to the medically uninsured. PMID- 11899625 TI - Pacing the standard of nursing practice in radiation oncology. AB - Radiotherapy frequently is used to treat both breast and lung cancers. Because of recent technological advances, radiation oncologists can deliver increasingly higher amounts of radiation to these areas. Inevitably, some patients requiring radiation therapy will have histories of cardiac disease and temporarily or permanently implanted cardiac pacemakers. Pacemakers are not immune to radiation; damage to their internal circuitry can occur if safeguards are not in place. A recent informal telephone survey of radiation oncology departments in Pennsylvania, as well as four nationally recognized cancer centers, yielded varied and inconsistent nursing practices in caring for these patients. A review of the literature found recommendations that were developed originally in 1989, then updated in 1994 and 1998, to be incomplete from a nursing perspective. Radiation oncology nurses must take a proactive role to deliver quality consistent care, while minimizing their liability, when working with this patient population. This can be achieved by implementing a nursing policy that incorporates the 10 recommendations described in this article. PMID- 11899626 TI - Hemochromatosis: a review. AB - Hemochromatosis is an autosomal recessive disorder that causes abnormal absorption of iron and results in iron overload. This disorder affects the liver, pancreas, heart, and endocrine systems, and if undetected and untreated, organ damage and death can result. The recent discovery of the gene associated with hemochromatosis has led to increased awareness of the disease and controversy over its diagnosis and treatment. Transferrin saturation is the best laboratory assay to detect the disease, and a liver biopsy is required to confirm a diagnosis. Current treatment involves therapeutic phlebotomy and iron-chelating agents. Information on diet modifications and genetic counseling for people affected with this disease should be included in patient education. PMID- 11899627 TI - Clinical evaluation of a positive pressure device to prevent central venous catheter occlusion: results of a pilot study. AB - One of the complications related to central venous catheters is occlusion secondary to thrombus formation within or surrounding the catheter lumen. Historically, methods to prevent these occlusions have included vigorous flushing, coordinated flushing-clamping techniques, and antithrombotic prophylaxis using low-dose warfarin or low molecular weight heparin. Positive displacement devices recently have become available that prevent retrograde blood flow and consequently reduce the risk of thrombus formation in the catheter lumen. Maintaining catheter patency results in fewer treatment delays and diagnostic procedures, decreased use of thrombolytics, lower costs, and increased patient satisfaction. A trial of a positive displacement device was conducted on an inpatient oncology unit to determine its effectiveness in preventing catheter occlusions. The easy-to-use device effectively reduced the number of occlusions and resulted in significant cost savings when compared to thrombolytic therapy. PMID- 11899628 TI - Recent developments in antiretroviral therapy. AB - This article reviews the antiretroviral medications in development as presented at the Eighth Annual Conference on Retroviruses and Opportunistic Infections. New medications for the treatment of HIV/AIDS are discussed by classification and a brief description of its unique quality is described. PMID- 11899629 TI - A serious look at the undertreatment of pain: part 11. PMID- 11899630 TI - Management of a widely disseminated skin rash. AB - T.J.'s case was interesting from the standpoint of both diagnosis and management. The recommended treatment for this drug reaction was prednisone: however, the use of a steroid in a patient who is neutropenic and has a fever is risky because the because the steroid can mask the symptoms of infection (e.g., fever). Administration of prednisone did help, and the patient experienced a rapid resolution of the skin rash. T.J. will need to avoid the use of these antibiotic agents in the future. Because it is unknown which antibiotic, vancomycin or ceftazidime, caused the allergic reaction, both medicines should be avoided. The decision to rechallenge a patient with a specific drug must be made on an individual basis. Rechallenging of a drug in patients who have had urticarial, bullous, or erythema multiforme-like eruptions can be very dangerous (Padial et al., 2000). Pinpointing the cause of a skin rash can be puzzling. Always ask the patient "Do you take any medicine for any condition (including aspirin, laxatives, vitamins, etc.)? Have you received any shots any shots in the last month?" Keep in mind that any chemical that is ingested can cause a cutaneous drug eruption. PMID- 11899631 TI - Bowel obstruction. AB - Bowel obstruction is a serious problem requiring prompt medical intervention to relieve intestinal distention, restore intravascular volume, and correct fluid and electrolyte balance. If these efforts fail, either curative or palliative surgical interventions become necessary. Oncology nurses are important members of the healthcare team caring for patients with bowel obstructions. Accurate nursing assessments are critical to monitoring the effects of these therapies and key to reducing the risk of preoperative and/or postoperative complications. PMID- 11899632 TI - Complementary and alternative dietary therapies. AB - Patients with cancer often feel that they can improve their physical condition and prognosis by altering what they eat. Their quest for nutritional information may lead them to alternative and complementary dietary therapies. Although many of thesetherapies appear to be "nontoxic" or "natural" they are often restrictive and place patients at risk for various complications and adverse events. Weitzman (1998) adds that prospective, randomized trials of alternative and complementary dietary therapies are needed, and that these nutritional therapies must be used with great caution by pediatric patients because this population is at high risk for nutritional deficiencies and diet-related complications. PMID- 11899634 TI - Interrater reliability of the Glasgow Coma Scale scoring among nurses in sub specialties of critical care. AB - The Glasgow Coma Scale (GCS) is used as an assessment tool to measure the levels of consciousness and coma in patients. This research investigated the reliability of scoring the GCS among registered nurses (RNs) working in five different sub specialty clinical areas of critical care; general intensive care, neurosurgical intensive care, coronary care, emergency room and post anaesthetic recovery room. Seven video recordings were made of six patients (one patient was recorded twice) having their level of neurological response assessed utilising the GCS. Seventy five RNs (15 from each sub-specialty) viewed each of the GCS assessments on the videotape and rated each patient on the scoring sheet provided. Analysis was performed for all RN subjects as a single group as well as separately for each of the five groups under investigation. The ratings for the first six videos were used to test interrater reliability and the scores from videotape four and seven (same patient) were used to calculate intrarater reliability. Based on comparison with expert scores, of the 75 participants, 38 responded correctly to eye opening responses; only 26 responded correctly to the motor response ratings. However, a better accuracy was achieved in the verbal response category with 67 participants responding correctly. Education qualifications and previous neurosurgical experience were statistically significant with regard to the nurses' accuracy of GCS assessment with p values of 0.004 and 0.043 respectively. The results were consistent with previously published studies demonstrating the motor response rating is most problematic in relation to rate accuracy. PMID- 11899635 TI - A consensus driven method to measure the required number of intensive care nurses in Australia. AB - This paper describes a methodology for determining the number of nurses required to staff Australia's intensive care unit (ICU) beds. The evidence used is level IV, that is the use of expert panel opinion, and it is the strongest and most accurate attempt yet to describe how Australia's ICU beds should be staffed with nurses. The researchers provide ratios of staffing applicable to a variety of situations that should be of use to ICU managers and hospital administrators. Equally, the broader calculations explaining the national supply and demand needs provide an easy to use approach and explanation suitable to health professionals, health administrators, policy advisors, governments, politicians and the broader community. Limitations of the approach and further recommendations are made to encourage future work in this area. Finally, a strong correlation between the number of available 'open' ICU beds in Australia and the number of nurses actually working in ICU at any given time is well demonstrated using the methodology outlined in this paper. Clarification of scope and terminology: This paper focuses on the nursing requirements of ICUs only; occasionally we use the word critical care nurse--this generally refers to those nurses who have completed a generic critical care nursing course but who, for the purposes of our study, are working in ICUs. Critical care units encompass ICUs but may also encompass recovery room, cardiothoracic units, coronary care, emergency departments and many other environments where critically ill patients are cared for and treated. This paper does not cover the broader scope of critical care units, only ICUs. PMID- 11899636 TI - The effects of nursing staff inexperience (NSI) on the occurrence of adverse patient experiences in ICUs. AB - Although many studies have attempted to define levels of staff experience appropriate for the care of critically ill patients, minimal data is available on the effect of nursing staff inexperience (NSI) on the occurrence of incidents in the intensive care environment. The Australian Incident Monitoring Study in Intensive Care Units (AIMS-ICU), an anonymous, voluntary incident reporting system, can help to identify problems in which NSI may be implicated. NSI may be an incident in itself or contribute to the occurrence of other incidents. The objective of this paper was to identify incidents associated with NSI and estimate their effect on the quality of patient care. Incidents related to NSI were extracted from the AIMS-ICU database and analysed using descriptive methodology. Seven hundred and thirty five reports covering 1,472 incidents were identified as relating to NSI. Of these, 282 were described in the narrative section and 453 were selected as a contributing factor by the reporter. Major categories for the 1,472 incidents included airway and ventilation (317) drugs and therapeutics (468), procedures, lines and equipment (219), patient environment (234) and unit management (234). An undesirable major adverse patient outcome was selected in 20 per cent of reports. NSI associated incidents reported to AIMS-ICU suggest that NSI can have a negative impact on the quality of care delivered to critically ill patients as shown by the occurrence and outcome of incidents related to such inexperience. Errors are more likely to occur when NSI is combined with staff shortage, inadequate supervision and high unit activity. When rostering or employing staff, nurse managers and educators must consider the special requirements of inexperienced nurses. Safe patient care requires these issues be included in discussions about ICU resource allocation. PMID- 11899637 TI - Quality use of medicines (QUM) in critical care: an imperative for best practice. AB - Quality use of medicines (QUM) as a discrete concept is gaining increasing importance in Australia and is supported by a policy platform which has federal government and health professional support. The QUM movement is also supported by a strong consumer base and this lobby group has been responsible for endorsement as a major health initiative. However, the importance of QUM to achievement of optimal patient outcomes has not achieved sufficient recognition in the critical care literature. Implicit in the discussion of QUM is the rational, ethical, safe and effective use of drugs within a best practice framework. Successful implementation of QUM requires appropriate infrastructure and the commitment and cooperation of medical, nursing and pharmacy staff. Support, education and training provide the prerequisites of knowledge, skills and awareness for quality use of medicines for all groups. An emphasis upon evidence based practice and the prevalence of polypharmacy in contemporary health care systems requires examination of factors that are barriers to best practice. QUM in critical care areas requires appropriately skilled staff who are competent to manage patients with a wide range of selected drugs, often in highly stressful situations. In many situations in critical care, the role of the critical care nurse is one of patient advocate. It is important to note that the delivery of critical care is not limited to a discrete setting and is inclusive of management at the trauma scene, assessment and delivery of care in the emergency department, through to intensive, coronary care and high dependency units. This paper presents a discussion of the concept of QUM and its relevance in the critical care context. Key theoretical, policy and research considerations for establishment of QUM in critical care are reviewed and discussed. This paper seeks to describe key issues in QUM and endorse the need for a research agenda in critical care. PMID- 11899633 TI - Alemtuzumab. PMID- 11899639 TI - The call for an evidence base--but what of the base? PMID- 11899638 TI - Making decisions: nursing practices in critical care. AB - This article reports the types and complexity level of decisions made in everyday clinical practice by critical care nurses. It also reports factors that influence the complexity of those decisions. A combination of methods were chosen for the two phase study. In the first phase, 12 qualified critical care nurses documented decisions (over a 2 hour period) on a clinical decision recording form designed by the researcher. In the second phase, participants attended a semi-structured focus group. From the analysis, five types of decisions were identified; assessment, intervention, organisation, communication and education. In addition to these documented decisions, three factors that influenced decision complexity were identified from a thematic analysis of the transcribed interviews; communication, patient related and properties of the decision. Nurses reported that communication decisions were the most difficult to make. However, the concept of nurses knowing the patient reduced the level of decision complexity. It is suggested that this has important implications for decision making practices of nurses working in the area of critical care and potentially for patient outcomes. PMID- 11899640 TI - Evidence based practice: are critical care nurses ready for it? AB - In the emergence of the evidence based practice movement, critical care nurses have struggled to identify scientific evidence on which to base their clinical practice. While the lack of critical care nursing research is a major concern, other important issues have significantly stalled the implementation of evidence even when it is available. A descriptive study of 274 critical care nurses was undertaken to examine nursing research activity in Victorian critical care units. The study aimed to identify critical care nurses' research skills, the barriers encountered in participation and implementation and the current availability of resources. Results revealed that 42 per cent of the nurses who participated in the study believed that they were not prepared adequately to evaluate research, and less than a third believed they were sufficiently skilled to conduct valid scientific studies. An association was found between nurses' ability to confidently perform research activities and higher academic qualifications. The study found that there is a lack of organisational support and management commitment for the development of evidence based nursing. In order to facilitate the implementation of evidence based practice, clinicians must be made aware of the available resources, be educated and mentored when carrying out and using clinical research, and be supported in professional initiatives that promote evidence based practice. It is argued that this will have positive implications for patient outcomes in the critical care environment. PMID- 11899641 TI - The future of clinical research in breast cancer: challenges. PMID- 11899642 TI - Benefit of the addition of paclitaxel to standard chemotherapy with 5 fluorouracil/doxorubicin/cyclophosphamide in patients with operable breast cancer. PMID- 11899643 TI - High-dose adjuvant chemotherapy for patients with high-risk breast cancer: efficacy is still an open question. PMID- 11899644 TI - High-dose chemotherapy with hematopoietic stem-cell transplantation for breast cancer: current status, future trends. AB - High-dose chemotherapy with hematopoietic stem-cell transplantation (HDC/HSCT) has been extensively studied as a potential treatment for breast cancer. A literature search of MEDLINE from January 1990 through December 1999 identified 497 published full papers. Of these articles, 120 reported the results of clinical trials, 78 were reviews, and 299 reported on issues related to the technology of peripheral stem cells, supportive care, and toxicity. The phase II data must be interpreted with caution, as it is subject to selection bias; transplant recipients tended to be younger, rigorously staged, and selected to be chemotherapy responsive. There continues to be controversy regarding the role of high-dose therapy in this disease. Only a few fully published randomized trials are available; these studies were powered only to detect large differences in survival and no benefit was shown. Several large controlled trials are either in progress or are too early for definitive analysis. This review analyzes the current literature on HDC/HSCT for breast cancer, identifying prognostic factors and discussing ongoing research designed to improve antitumor effects. PMID- 11899645 TI - Exemestane: a novel aromatase inactivator for breast cancer. AB - Exemestane is a unique inactivator of the aromatase enzyme and differs from the two approved aromatase inhibitors. It is well absorbed at a daily oral dose of 25 mg and produces significant suppression of aromatase and plasma estrogen levels without androgenic side effects. Toxicity is mild with menopausal symptoms predominating. Exemestane is approved for the treatment of postmenopausal women with recurrent breast cancer. In reported clinical trials, exemestane was effective in patients failing tamoxifen, megestrol acetate, or even other aromatase inhibitors in phase II trials and was superior to megestrol acetate in a phase III randomized trial in which an early survival advantage for exemestane was observed. Studies evaluating first-line exemestane for adjuvant use and as a chemopreventive agent are underway. PMID- 11899646 TI - Clinical relevance of occult metastatic cells in the bone marrow of patients with different stages of breast cancer. AB - Data are emerging about the prognostic relevance of occult metastatic cells in the bone marrow of patients with various solid tumors. Discrepancies among different studies on the prognostic relevance of isolated tumor cells may be caused by tumor cell heterogeneity and the use of different immunoassays. There is increasing evidence that validated anticytokeratin antibodies (e.g., A45-B/B3) represent the present standard for the detection of isolated tumor cells. This immunocytochemical assay allows the identification of patients with occult tumor cell dissemination that cannot be identified by conventional screening methods in tumor staging. According to recent studies, these patients are at higher risk for subsequent development of distant metastases and might therefore benefit from early systemic therapy. At advanced stages of the disease, the micrometastatic tumor load after adjuvant therapy, or at the time of emerging recurrences, appears to reflect the tumor's ability to progress. Therapeutic monitoring and cell-cycle independent antibody-based therapy are among possible implications of this new, promising diagnostic tool. The present review also focuses on state of the art, reliable detection methods of occult metastatic cells in the bone marrow of breast cancer patients and on the prognostic relevance of these cells at different stages of the disease. PMID- 11899647 TI - Phase II trial of gemcitabine/doxorubicin/paclitaxel administered every other week in patients with metastatic breast cancer. AB - The present trial was designed to determine the efficacy of the combination of gemcitabine/doxorubicin/paclitaxel (GAT) delivered every other week as first-line therapy in patients with metastatic breast cancer. From February 1998 to September 1999, 41 patients were included in this trial. Doses delivered were doxorubicin 30 mg/m2 on day 1 and paclitaxel 135 mg/m2 plus gemcitabine 2500 mg/m2 both given on day 2, every 14 days. Doses were selected from a previous phase I trial conducted at our institution. Eligibility criteria for the phase II trial included histologically confirmed metastatic breast cancer with bidimensionally measurable lesions; no prior therapy for metastatic disease; adjuvant or neoadjuvant chemotherapy was allowed if given more than 1 year before and cumulative doses of doxorubicin or epirubicin were less than 200 mg/m2 or 360 mg/m2, respectively; Eastern Cooperative Oncology Group (ECOG) performance status of 2 or less; and adequate hematological, hepatic, and renal function. Prophylactic use of granulocyte colony-simulating factor (G-CSF) was allowed if patients were not fully recovered (absolute neutrophil count greater than 1500/microL) from chemotherapy administration before the next dose. Left ventricular ejection fraction was determined initially, at the end of the study, and every 6 months thereafter. The patients' median age was 55 years (range, 33 68 years), and their median ECOG performance status was 0 (range, 0-1). Twenty eight patients had received adjuvant therapy, 17 with epirubicin (none with doxorubicin). Metastases were present in the bone (19 patients), lung (19 patients), liver (11 patients), and soft tissues (18 patients). Twenty patients had one metastatic site and 21 had two or more sites. Efficacy was assessed on an intent-to-treat basis. A total of 216 cycles of GAT were given. Twenty-two percent of the courses were delayed or given at reduced doses mostly due to neutropenia or thrombocytopenia. G-CSF was required in 58% of the cycles. Grade 3/4 neutropenia was the main toxicity and appeared in 17 patients, one of whom had an episode of febrile neutropenia. Nonhematological toxicities consisted mainly of neurotoxicity and myalgias. A drop of 10%-20% in the left ventricular ejection fraction was detected in two patients and another patient had a decrease greater than 20%, although none developed symptoms of heart failure. Overall response rate was 80.4% (95% confidence interval: 68.3-92.5), with 15 patients (36.6%) achieving a complete response. Median survival time was 27 months and median time to progression was 15 months. The GAT combination is feasible and very active in patients with metastatic breast cancer, with an encouraging response rate including a high rate of complete responses. No congestive heart failure was documented and other toxicities were mild, with the exception of neutropenia. PMID- 11899649 TI - Cytopathology of metastatic breast cancer. AB - A 52-year-old woman developed an infiltrating ductal carcinoma of the breast 16 years after being treated for comedocarcinoma of the left breast. Although the tumor was high grade with lymphatic space invasion and incompletely excised, the patient declined adjunctive therapy. Within 2 years, she developed metastasis to supraclavicular lymph nodes, which was diagnosed by fine-needle aspiration cytology. PMID- 11899648 TI - A study of the value of p53, HER2, and Bcl-2 in the prediction of response to doxorubicin and paclitaxel as single agents in metastatic breast cancer: a companion study to EORTC 10923. AB - This study assesses the potential value of the tumor markers p53, HER2, and Bcl-2 in predicting the clinical response to doxorubicin and paclitaxel as single agents in the treatment of metastatic breast cancer. The primary tumors of 114 patients in the European Organization for Research and Treatment of Cancer 10923 trial were assessed by immunohistochemistry using monoclonal antibodies; the results were correlated with clinical response to therapy. HER2 was positive in 24% of patients, p53 was positive in 25% of patients, and Bcl-2 was positive in 49% of patients. There was no correlation between the expression of any of the markers and the clinical response to either agent. Although methodologically limited, this study does not support the use of p53, HER2, or Bcl-2 to assist the selection of anthracycline versus taxane in metastatic breast cancer. PMID- 11899650 TI - Comprehensive review of sentinel lymphadenectomy in breast cancer. AB - Sentinel lymph node dissection (SLND) is a minimally invasive technique to stage axillary lymph nodes in breast cancer. The complications associated with SLND are minimal, especially when compared to routine axillary lymph node dissection (ALND), and it can be performed with an overall identification rate of greater than 90% and a false-negative rate less than 5%. Despite this, SLND is not ready to replace routine axillary dissection, since we have no long-term results for these patients. What the clinical recurrence rates will be in women who undergo SLND only and how that will translate into survival rates has yet to be discovered. SLND is also a difficult technique to perform, as documented in the early SLND studies. It is imperative that each individual surgeon perform a series of cases in which SLND is combined with immediate ALND, so that identification rates and false-negative rates can be determined. Once a track record of successfully performed SLND has been established, SLND can be solely used for node-negative women. It is strongly recommended that all surgeons join one of the National Cancer Institute (NCI)-sponsored clinical trials for SLND in early breast cancer, so that many of these questions concerning SLND can finally be answered. PMID- 11899651 TI - Prophylactic surgery for women at high risk for breast cancer. AB - Women at high risk for the development of breast cancer have several options open to them including increased cancer surveillance, prophylactic mastectomy and/or oophorectomy, and chemoprevention. We consider high-risk women to be those with known BRCA mutations or a strong family history characterized by multiple relatives with breast cancer, early age at diagnosis, and in some families, ovarian cancer. We present existing data regarding prophylactic surgery for these women. Essentially, a woman at high risk for breast cancer may choose to undergo bilateral prophylactic mastectomy, with or without reconstruction. For patients who have a known breast cancer, contralateral mastectomy is also an option. Finally, for women in families with a strong incidence of ovarian cancer, prophylactic oophorectomy can be considered. PMID- 11899652 TI - Paclitaxel, 5-fluorouracil, and leucovorin (TFL) in the treatment of metastatic breast cancer. AB - To assess the activity of paclitaxel in combination with 5-fluorouracil (5-FU) and leucovorin in breast cancer, a phase II trial was conducted in women with metastatic disease. Toxicity, response rate, median survival, median duration of response, and median time to disease progression were measured. Between January 1994 and May 1996, 47 patients with metastatic breast cancer and an Eastern Cooperative Oncology Group (ECOG) performance status (PS) < or = 2 who had previously been treated with chemotherapy received 175 mg/m2 paclitaxel over 3 hours on day 1. After paclitaxel administration, 300 mg intravenous (i.v.) leucovorin over 30 minutes was administered followed by 350 mg/m2 i.v. push 5-FU. Both 5-FU and leucovorin were given on days 1-3. Treatment was repeated every 28 days for a minimum of 6 cycles per patient. Two (4%) patients had a complete response and 21 (45%) patients had a partial response for an overall response rate of 49% (95% confidence interval: 35%-63%). The median survival was 17.7 months, median duration of response was 8.6 months, and median time to disease progression was 6.3 months. There was no statistical difference in survival or time to progression between anthracycline-naive, anthracycline-sensitive, and anthracycline-resistant patients. Nine (19%) patients had grade 3 or 4 neutropenia, and no patient required blood or platelet transfusion. The most frequently observed nonhematologic toxicities were arthralgia and myalgia. Pharmacokinetic data were obtained on 19 patients. Responders had higher peak plasma concentrations of paclitaxel than nonresponders (4.46 vs. 2.9 micrograms/mL; P = 0.02). Paclitaxel/5-fluorouracil/leucovorin is an active, well tolerated regimen for patients with metastatic breast cancer. PMID- 11899653 TI - Internet usage among women with breast cancer: an exploratory study. AB - An increasing number of breast cancer patients are accessing the Internet for medical information. A survey was administered to breast cancer patients and their families attending follow-up outpatient clinics in a comprehensive cancer care center to explore their frequency of Internet use, their motivation for online activity, the type of information they sought, and the perceived impact of the information they found on the Internet on their medical care. The survey was conducted over a 4-month period. A total of 107 surveys were returned. Seventy nine of these (74%) were from patients while 28 (26%) were from family members and friends. Thirty-four of the patient responses (43%) indicated that the patient had used the Internet to look for cancer-related information. Patients who had used the Internet to access cancer-related information were significantly younger (P = 0.007), better educated (P = 0.027), and less satisfied with the amount of treatment-related information given by caregivers than those patients who had not used the Internet to access cancer-related information (P = 0.032). The majority of patient Internet users desired more information on their cancer and its treatment (91%), looked up information that was presented to them by their clinicians (66%), researched other treatment options (63%), and obtained more information on "alternative treatments" (63%). Patient Internet users generally found the cancer-related information on the Internet to be useful, and the majority discussed Internet-derived information with their health care providers and perceived that clinicians listened to such information. However, 53% were undecided about the trustworthiness of the medical information obtained via the Internet. Internet nonusers commonly lacked Internet access (53%) or were unfamiliar with the Internet (33%), but few (13%) distrusted Internet-derived information. This exploratory study underscores the need for more research in this area, specifically with the aims of identifying and verifying factors that lead patients to use the Internet and the impact of their online activities on their medical care. PMID- 11899655 TI - Use of taxanes in the treatment of patients with breast cancer has increased dramatically over the past 8 years. PMID- 11899656 TI - The EORTC strategy for the early 2000's. PMID- 11899654 TI - Role of 2-[18F]-fluorodeoxyglucose (FDG) positron emission tomography (PET) in the early assessment of response to chemotherapy in metastatic breast cancer patients. AB - We investigated the role of 2-[18F]-fluorodeoxyglucose (FDG) positron emission tomography (PET) in the early evaluation of response to chemotherapy in metastatic breast cancer patients. Breast cancer patients who received an epirubicin/paclitaxel--containing regimen as first-line treatment for metastatic disease were included in this study. A PET study was performed within 1 week before the start of treatment, at day 8 after the first course, and at the end of the planned program of chemotherapy. Tumor response was determined clinically and radiographically every 2 courses of treatment. Thirteen patients with metastatic breast cancer who were referred for treatment protocols with gemcitabine/epirubicin/paclitaxel or epirubicin/paclitaxel chemotherapy regimens were included in this study. All metastatic sites were easily visualized on the baseline FDG-PET images, obtained 50 to 60 minutes after tracer injection. Nine patients who completed the planned courses of chemotherapy and the FDG-PET studies were available for analysis. In the six patients who achieved a response to treatment, median glucose standard uptake value (SUV) (semiquantitative analysis) was 7.65 (range, 3.4-12.3) at baseline, 5.7 (range, 2.8-7.6) at day 8 after the first course, and 1.2 (range, 0.99-1.3) at the end of the 6 planned courses of chemotherapy. Three patients who obtained a stable disease as best response had no significant decrease in tumor glucose SUV compared to baseline levels. Qualitative visual analysis in the six responding patients showed a decrease in delineation of tumor mass from background activity soon after the first course, while the nonresponding patients had no significant modification from basal levels. Semiquantitative FDG-PET scanning of metastatic breast cancer sites showed a rapid and significant decrease in tumor glucose metabolism soon after the first course of treatment in patients who achieved a response to first line chemotherapy. On the contrary, no significant decrease was observed in nonresponding patients. PMID- 11899657 TI - The future of clinical research in breast cancer: opportunities. PMID- 11899658 TI - [The variation of two types of spatiotemporal organization of brain potentials at different stages of a cognitive set based on illusory perception of length (Muller-Layer illusion)]. AB - The spatiotemporal organization of brain potentials (STOP) under conditions of their multichannel recording was studied was studied in 35 young healthy subjects during formation, actualization, and extinction of the cognitive set on the basis of illusory perception of length (Muller-Layer illusion) by means of quantitative comparison of successive electroencephalotopograms (ET). The ET is an array of momentary values of potentials simultaneously recorded in all the derivations. An ET is characterized by a relief and mean level. Time changes in these characteristics determine a certain type of cortical STOP. Two main types of STOP were revealed. The interaction between these two types of STOP depended on the kind of performance of a subject and was ferlected in the coefficient of dynamic variation (CDV) of ETs. The CDVs were significantly different in subjects with stable and unstable forms of the cognitive set; in the latter this value was higher. The spectral characteristics of the EEG and STOP dynamics were compared. Analysis of power stectra revealed the significant intergroup differences in the alpha 1 frequency range both in the EEG and STOP; the power of this frequency range was higher in subjects with the stable set. PMID- 11899659 TI - [Retention of a memory trace of learning induced by a new and extinctive stimulus in C57BL/6J and BALB/c mice]. AB - Influence of preliminary habituation to the experimental box on passive avoidance learning was studied in mice of C57BL/6J and BALB/c strains. The 20-fold preexposure of mice to the environmental stimulus inhibited the acquisition and consequent retention of a memory trace in BALB/c mice and did not affect the acquisition in C57BL/6J mice. Subsequent retention in this strain was facilitated. The interstrain differences are discussed with respect to detected genetically determined types of behavior and exstinctive inhibition at the habituation stage. PMID- 11899660 TI - [The multifactor method of EEG separation into the cortical and deep components]. AB - A new method of separation of multichannel brain electrical activity into cortical and subcortical components with the help of multifactor analysis is proposed. The method provides a means for isolation and, consequently, more reliable localization of sources of electrical activity not only in deep brain structures (on the basis of the dipole model) but also on the cortical surface. The proposed method does not depend on the rotation and interpretation of factors, and no data losses occur. The mufasel algorithm is based on integration of all selected factors (within a particular EEG or EP time segment) in two groups using a statistical criterion, which defines general and specific factors. It is assumed that general factors loaded with highly correlated derivations predominantly describe the electrical activity of deep brain structures, whereas specific factors loaded by the dynamics of electrical activity in individual derivations, reflect the integrated activity of cortical brain structures. PMID- 11899661 TI - [Evaluation of the role of the adaptive result in the theory of the functional systems]. AB - The fundamental of the theory of the functional systems, i.e., the concept of the useful adaptive result as a universal system-forming factor is considered. It is suggested that the adaptive result is not system-forming in behaviors actualized exclusively due to activity of systems developed earlier. It is argued that positive mutations may serve as the system-forming factor for hereditary determined behavioral forms. In all other cases of goal-directed behavior (except conditioning) the aim of performance as a model of the future result plays the decisive role. Only in conditioning the classical concept of the system-forming role of the adaptive result seems to be undeniable. The refined ideas about the mechanisms of formation of the functional systems may be useful in analysis of a number of animal and human functions (learning, emotional stress, neuroses, etc.). PMID- 11899663 TI - [Gender distinctions in recognition of partly masked geometric figures]. AB - Visual recognition of entire and partly masked geometric figure by men and women was studied. Probabilities of correct recognition of the entire (control) and partly masked figures (without some parts of their lines or corners) under conditions of their near-threshold tachistoscopic presentation were compared. A gradual decrease in recognition probability with increasing masking was observed. It was more pronounced for figures without some corners than for figures without part of their sides. Reliable gender differences were found: men better than women recognized figures, especially, with masked corners. Possible reasons for gender differences in recognition of geometrical figures are discussed, in particular, the role of striate sensitivity to a single light bar and a line crossing in the observed phenomena. PMID- 11899662 TI - [Characteristics of motor and cardiac activity in men during the performance of test "shooter"]. AB - Subject's behavior and electrocardiogram were investigated during execution of the test "Shooter". Three kinds of strategies were used by subjects during the task performance. Three types of the motor activity of subjects were also found in the "interresulting" periods during the system activity. The spatial and temporal characteristics of these types of activity were similar to those during the actual achieving the result. Differences in the dynamics of the R-R-interval changes were found in the periods before achieving the positive or negative result. PMID- 11899665 TI - [Chaotic nonlinear dynamics of prestimulus EEG and the structure of auditory evoked potentials ]. AB - The dynamic pattern of a prestimulus EEG was studied by the fractal analysis technique. The character of this pattern was shown to affect the structure of cortical auditory evoked potentials (EP). A monoperiodic pattern in the control frequency band was accompanied by a formation of several new attractors with low dimensionality attributed to simultaneous functioning of several weakly connected dynamic systems with an indefinite trend of a leading process. During development of multiperiodic processes in EEG segments, the fractal analysis revealed a tendency for a formation of a single complex dynamic system with high dimensionality of the attractor. Characteristic changes in parameters of the primary and middle-latency EP components, their correlation, and factor models are related with the character of nonlinear patterns. Analysis of variance revealed the most effective role of pattern changes in the EEG alpha control band. The structure of combination of parameters of the EP primary components into connected complexes depends on the nonlinear prestimulus EEG patterns. The above predictors determine differently directed and differently pronounced changes in parameters of the middle-latency positive waves. PMID- 11899664 TI - [A correlation between the efficiency of probabilistic prediction and characteristics of left and right interhemispheric activation]. AB - The interhemispheric difference in the level of EEG activation was studied as a correlate of the efficiency of human probabilistic activity. The interhemispheric asymmetry of the arousal duration in the projection (occipital) and associative (central) areas was assessed in two tested groups of subjects: with an adequate prediction and difficulties in predictive activity under conditions of different information significance of a stimulus. Under conditions of relevant stimulation, the asymmetry coefficient was higher, and the desynchronization reaction in the central areas of the left hemisphere was considerably shorter in bad predictors than in good ones. It is suggested that the asymmetry observed in bad predictors is determined by a nonspecific activation of subcortical structures, and in good predictors the asymmetry is related with the local neocortical activation. PMID- 11899666 TI - [Individual typological characteristics of higher nervous performance and asymmetry of hippocampus and amygdala electrical activity in dogs]. AB - Electrical activity of the frontal cortex, dorsal hippocampus, basolateral amygdala and lateral hypothalamus of both hemispheres was recorded in nine dogs in the state of quiet wakefulness without any stimulation. Individual typological features of higher nervous activity were assessed by the animal performance under conditions of free choice of the reinforcement mode: either high probable but of low alimentary quality, or with low probability but more valuable. Mean values of the maxima of crosscorrelation function between electrical activity of the investigated structures of two hemispheres were used as a basis for assessment of conditions for interaction between left and right formations. For the hippocampus and amygdala, in some dogs these conditions were the best in the theta and beta 2 ranges, in other animals--in the theta and alpha bands. In phlegmatic dogs, spectral densities in the theta range were higher in the left hippocampus than in the symmetrical structure, in sanguine animals spectral densities in the theta and beta 2 ranges in the hippocampus and amygdala were higher in the right hemisphere than in the left one. Thus, the hemispheric asymmetry of electrical activity of the limbic formations seems to be an important factor, which determines the individual typological features of the higher nervous activity in dogs. PMID- 11899667 TI - [Nature of functional motor asymmetry in animals: state of the problem]. AB - Handedness in skilled movements of animals is a result of interaction of innate motor preference and learning. The nature of the innate preference is not clear. Breeding of right-handed and left-handed mice revealed that the degree rather than direction of motor preference is an inherited feature. There is, however, a correlation between the direction of preference and a number of morphological, functional, and neurochemical characters. Shifts of a preference direction were found in some strains of mice. Differences between right-handed and left-handed rats were revealed in social behavior, learning, and resistance to forced retraining. Strains of rats with different forms of genetic epilepsy were characterized by the predominance of animals with a certain direction of the motor preference. This evidence suggests some genetic influence on a direction of the motor preference. Perhaps, genetic and environmental factors closely interact in determining motor preference in animals. PMID- 11899668 TI - [Instrumentalization of movements induced by stimulation of motor cortex with food reinforcement in dogs]. AB - Contrary to some literature data, the possibility to instrumentalize the movements (liftings) of the hind limb elicited by stimulation of the corresponding contralateral area of the motor cortex was shown. The instrumental reflex (spontaneous high lifting of the hind limb) was acquired after a number of uniform trials: cortical stimulation--movement--food. Food delivery was preceded by a click, which was presented during the hind limb lifting and served as a secondary reinforcement. The acquisition was rather prolonged (50-200 trials) and demanded some special conditions. The results count in favor of the viewpoint that the motor cortex can directly participate in establishing the instrumental conditioned connection (motivation--movement), and simple instrumental movements can be initiated through this connection. PMID- 11899669 TI - [Experimental analysis of systemogenesis process: c-fos gene expression in chicken brain during stimulation of development of a species-specific acceptor of action response]. AB - The aim of this study was to localize the areas of the chick brain involved in the development of filial predisposition to follow conspecifics. Expression of the immediate early gene c-fos mRNA was used to map such structures. One-day-old chicks were stimulated (primed) by placing for 90 min into a running wheel in darkness or by exposure to a loud nonspecific sound for 180 min. Brains of a part of these chicks taken 45 min after the beginning of priming were used for c-fos expression study. The remaining chicks were tested 24 hours after priming by simultaneous presentation of a "natural" object (stuffed fowl) and an "artifical" object (rotating red cube). Primed chicks demonstrated significant preference for the natural object as compared to untreated control chicks. In the forebrain of chicks primed both in the running wheel and by acoustic stimulation a significant c-fos expression was found in the medial part of the caudal neostriatum. Priming in the running wheel additionally induced c-fos expression in the lobus parolfactorius, while priming by acoustic stimulation produced high c-fos expression in the archistriatum. Both these areas are known to be involved in filial imprinting. The results suggest that these structures can be also involved in the development of filial predisposition in chicks. PMID- 11899670 TI - [Distinctions in metabolism of serotonin and dopamine in rat brain during latent inhibition]. AB - Latent inhibition (LI) is a behavioral phenomenon, in which repeated presenting of a non-reinforced stimulus retards conditioning to this stimulus when it is coupled with a reinforcer. In order to find specific serotonin (5-HT- and dopamine (DA) changes mediating the LI, the 5-HT and DA metabolism was investigated in certain brain regions. Oxidative deamination of 5-HT and DA by monoamine oxidase (MAO) was determined in the prefrontal cortex, striatim, amygdala, and hippocampus at preexposure and testing stages of the LI using the passive avoidance procedure in rats. Preexposed animals demonstrated high MAO activity for 5-HT deamination in the amygdala and striatum and lower MAO activity for DA deamination in the amygdala and hippocampus. After testing the LI, a high level of 5-HT deamination by MAO was revealed in the amygdala, white the lower level of 5-HT deamination by MAO was shown in the prefrontal cortex. At the same time, no changes in DA metabolism were found in all the brain regions studied. Thus, the role of dopaminergic system in the LI effect may be limited by the preexposure stage. The obtained evidence suggests that the enhanced 5-HT activity in the amygdala and striatum induced by the preexposed stimulus is a principal biochemical mechanism underlying the LI. PMID- 11899671 TI - [Concerning a conflict nature of the "spatial delayed response" test]. AB - Neuropsychological analysis of rats' performance of the spatial delayed response (SDR) in different testing conditions revealed a conflict nature of the indirect variation of the SDR task. It was found that the execution of the response based on the image short-term memory interferes with the response differentiation acquired during learning the rule of indirect SDR performance, i.e., during acquisition of the spatial discrimination. It is evident that the maximization of conditions, which promote the acquisition of response differentiation (additional training of animals for spatial discrimination), makes it difficult to perform the indirect variation of the SDR task, while the minimization of these conditions facilitates the correct task performance. PMID- 11899672 TI - [Involvement of rabbit motor cortex neurons in the instrumental behavior before and after chronic ethanol consumption: a comparison with the limbic cortex]. AB - The effect of acute administration is significantly more prominent in the limbic (cingulate) than in the motor cortex [9]. We proposed that the limbic cortex is more sensitive also to chronic ethanol treatment (CET). It was shown that morphology as well as neuronal activity of the limbic cortex changed greatly after the CET [7]. The missing link of testing the above proposition was a comparison of the obtained data with the results of the experimental study of CET influence on the motor cortex. Morphology of the anterolateral motor cortex and activity of its neurons in the instrumental food-acquisition behavior were studied in 6 male rabbits after CET (9 months). It was found that the limbic cortex was modified morphologically and functionally to a significantly greater extent than the motor cortex. We consider the fact that in the limbic cortex of a healthy individual there are many neurons, which for a while cease their discharges after the acute ethanol administration, to be among the most important reasons for this difference. Such-like repeated activity interruptions in the course of CET impair the performance of the systems incorporating these neurons. In such a way ethanol prevents all neurons, especially the mentioned ones, from receiving adequate metabolic supply that is necessary for their survival and functioning. PMID- 11899673 TI - [Chronic stress and forming of psychoemotional state during prepubertal period in rats]. AB - Immediate and long-lasting effects of chronic stress during prepubertal period (21-32 postnatal days) on anxiety- and depression-related behavior were studied in Wistar and ISIAH (inherited stress-induced arterial hypertension) rats. Significant interstrain differences were found. Both juvenile and adult ISIAH rats were less anxious in the elevated plus-maze and less depressed in the forced swimming test. Immediate effects of the prepubertal stress were similar in both rat strains and depended on the type of stimulation. Long-lasting effects were genotype-dependent. Chronic prepubertal handling exerted an anxiolytic effect in young ISIAH and Wistar rats and adult Wistar rats. Immediate anxiogenic effect of prepubertal unpredictable stress was preserved only in adult ISIAH rats. Depression-related behavior was intensified by the unpredictable stress in young animals, whereas the long-lasting effect was observed only in adult hypertensive rats. PMID- 11899674 TI - [Mature bone radionecrosis: from recent physiopathological knowledge to an innovative therapeutic action]. AB - Osteoradionecrosis is a severe radiotherapy (RT) injury by healing failure, late effect and spontaneously irreversible by tissue death. Histologically, it consists in a pagetoid mosaic that combines a defective osteogenesis with an osteoclastic osteolysis and more marginally an osteolytic osteolysis, turned to account to fibroblastic and collagenic fibrosis. Several pathogenic hypotheses favor sometimes a vascular hypoxic hypotheses, sometimes a fibro-atrophic hypothesis. Various events start up or favour ORN as traumatisms (dental extraction, surgery,...) or bacterian infection on fistula. In clinic, adult mature bone concerned is the mandible after head and neck RT by septic ORN, and the hip after pelvic RT by aseptic ORN. For each, epidemiology, clinic and therapeutic aspects are developed. Usual therapeutic attitudes consisted in restriction of defavorable associated events (dental extraction, infection, RT dose, chemotherapy,...) and devitalized tissue removal. Physiopathological therapeutic innovatives aspects are proposed to struggle against radiation induced fibrosis associated and to limit bone destruction. PMID- 11899675 TI - [First surgery followed by vaginal curietherapy in small-volume uterine cervix cancer: an alternative to the association of uterovaginal curietherapy and surgery]. AB - PURPOSE: Evaluate the results of the treatment of small uterine cervix cancer with the association of surgery and postoperative vaginal brachytherapy, without unfavourable prognostic factors. PATIENTS AND METHODS: After radical hysterectomy with lymphadenectomy, 29 women (mean age: 44 years) with carcinoma < 25 mm (26 stage IB1, 3 IIA, mean size: 15 mm) were treated by post-operative prophylactic vaginal brachytherapy using low dose rate. Ovarian transposition was performed at the surgical time in 14 young women (mean age 35 years). RESULTS: The actuarial specific survival rates at 5 and 10 years were 100% and 90% respectively, with a mean follow-up 75 months. Only one local recurrence was observed. The rate of grade 1 post-operative complication was 7%. The conservation rate of the ovarian function was 85% for young women. CONCLUSION: Treatment of small volume uterine cervix cancer using first surgery and post-operative vaginal brachytherapy is a reliable therapeutic option. The results in terms of specific survival and complications are the same with those after standard association of preoperative uterovaginal brachytherapy and surgery. PMID- 11899676 TI - Monitoring of therapy in head and neck patients during the radiotherapy by measurement of Cyfra 21-1. AB - PURPOSE: Cyfra 21-1, measuring serum fragments of cytokeratin 19, has been found to be related to tumour stage and tumour size in patients with cervical cancer. It could be a promising marker in squamous lung cancer. We evaluated this new marker with carcinoembryonic antigen, (CEA) and squamous cell carcinoma antigen (SCC-Ag) in the monitoring of 27 patients with head and neck cancer. PATIENTS AND METHODS: The retrospective study group consisted of 27 patients, 17 not suited for surgery and 10 after laser resection. Patients were clinically staged according to the TNM-classification. The mean age of the patients was 53 years (range 37-70 years). Serum levels of each marker were studied in relation to tumour stage and clinical status of the patients during radiotherapy and 6 weeks after the end of the treatment. The clinical performance of the various assays to separate those patients with complete remission from those patients with the presence of tumour was assessed. RESULTS: Pre-treatment serum Cyfra 21-1, CEA, and SCC-Ag levels were not related to stage of disease and were not found to be predictive of tumour response. The clinical performance of post-treatment serum SCC-Ag levels in predicting the presence of tumour was not better than the Cyfra 21-1 assays. CONCLUSION: We could not conclude from this study that Cyfra 21-1 marker is an additional parameter in identifying patients at risk of residual tumour after treatment, recurrent or progressive disease. An elevation of cyfra 21-1 marker was not detectable in 70% of the cases with macroscopic tumour. Therefore, Cyfra 21-1 is not a reliable parameter for the monitoring of patients with head and neck cancer during radiotherapy. PMID- 11899677 TI - [Conformal therapy of locally advanced cholangiocarcinoma of the main bile ducts]. AB - PURPOSE: Retrospective study of 23 patients treated with conformal radiotherapy for a locally advanced bile duct carcinoma. PATIENTS AND METHODS: Eight cases were irradiated after a radical resection (R0), because they were N+; seven after microscopically incomplete resection (R1); seven were not resected (R2). A dose of 45 of 50 Gy was delivered, followed by a boost up to 60 Gy in R1 and R2 groups. Concomitant chemotherapy was given in 15 cases. RESULTS: Late toxicity included a stenosis of the duodenum, and one of the biliary anastomosis. Two patients died from cholangitis, the mechanism of which remains unclear. Five patients are in complete remission, six had a local relapse, four developed a peritoneal carcinosis, and six distant metastases. Actuarial survival rate is 75%, 28% and 7% at 1, 3 and 5 years, respectively (median: 16.5 months). Seven patients are still alive with a 4 to 70 months follow-up. Survival is similar in the 3 small subgroups. The poor local control among R0N+ cases might be related to the absence of a boost to the "tumor bed". In R1 patients, relapses were mainly distant metastases, whereas local and peritoneal recurrences predominated in R2. CONCLUSION: Conformal radiochemotherapy delivering 60 Gy represents a valuable palliative approach in locally advanced biliary carcinoma. PMID- 11899678 TI - [Radiotherapy of lung cancer: the inspiration breath hold with spirometric monitoring]. AB - A CT acquisition during a free breathing examination generates images of poor quality. It creates an uncertainty on the reconstructed gross tumour volume and dose distribution. The aim of this study is to test the feasibility of a breath hold method applied in all preparation and treatment days. Five patients received a thoracic radiotherapy with the benefit of this procedure. The breathing of the patient was measured with a spirometer. The patient was coached to reproduce a constant level of breath-hold in a deep inspiration. Video glasses helped the patients to fix the breath-hold at the reference level. The patients followed the coaching during preparation and treatment, without any difficulty. The better quality of the CT reconstructed images resulted in an easier contouring. No movements of the gross tumour volume lead to a better coverage. The deep breath hold decreased the volume of irradiated lung. This method improves the reproducibility of the thoracic irradiation. The decrease of irradiated lung volume offers prospects in dose escalation and intensity modulation radiotherapy. PMID- 11899679 TI - [Non-Hodgkin's lymphoma of the uterus: apropos of 4 cases and review of the literature]. AB - The primary non hodgkin's lymphoma of the uterus is rare. This rarity explains of one part certain difficulties of the histological diagnosis and on the other hand the absence of a therapeutic strategy clearly established. We report 4 cases of primary non-hodgkin lymphoma of the uterus. Two patients had a cervical location, the two other had corpus location. The average age of our patients is of 59 years (extremes: 54-68). Histological diagnosis was confirmed by biopsy for the cervical location. For the corpus location, it is study of the uterus after hysterectomy which retained the diagnosis of lymphoma. The type of the lymphoma was low grade in two cases and high grade in the two other cases. The disease was limited to the pelvis for all our patients (stage IE according to Ann-Arbor's classification). The treatment consisted of an association of chemotherapy and radiotherapy in both cases of lymphoma of the cervix and in a radical hysterectomy followed by chemotherapy for the two cases of lymphoma of the corpus. Our patients are regularly followed, with an average follow-up of 56 months. Two patients are in disease free, the third patient presented a dissemination of the disease and the fourth patient presented a squamous cell carcinoma of the lung. PMID- 11899680 TI - [Will there be an alternative to scheduled death of radiotherapy in French speaking Africa?]. PMID- 11899681 TI - [Report on the 43rd Congress of the American Society for Therapeutic Radiology and Oncology (ASTRO), San Francisco, 4-8 November 2001]. PMID- 11899682 TI - [Nicotine substitution therapy: 10 years later]. AB - To be efficient, nicotine substitution therapy (NST) must be part of a global strategy comprising several stages. The first consists in evaluating and re enforcing motivation, since smoking cessation is impossible unless the patient is clearly motivated. The second stage is the withdrawal period that lasts for several days or months. NST is the first medicinal treatment that has demonstrated its efficacy in controlled studies: it reduces the withdrawal syndrome and doubles the chances of success at the end of the standard 3-month treatment. Recent studies have shown that the results can be improved: by adapting the dose to the degree of dependency and by using higher doses and/or associating two types of nicotine substitutes; by prolonging NST for as long as the withdrawal syndrome persists; by treating the anxiety and depression, often present in heavy smokers. The third stage is aimed at avoiding relapses, by ensuring prolonged follow-up and treating the various possible causes: eating disorders and weight gain, acute or chronic stress, depression, environment.... At all the stages of care, behavioural and cognitive therapy enhances the chances of success. PMID- 11899683 TI - [Synercid emergency prescription program. The French experience]. AB - OBJECTIVE: An emergency-use program forof Synercid (quinupristin/dalfopristin, Q/D) has been set up following the occurrence of Gram-positive infections with no therapeutic alternatives to the available antibiotic arsenal. METHODS: The experience in France is based on a collective of 88 infections analysed in 74 patients. The most frequent clinical indications were: central catheter-related bacteremia, bone and joint infection, endocarditis and, intraabdominal infection. The most frequently causative pathogens were: S. aureus (n = 26, including 24/26 meticillin-resistant), coagulase negative staphylococci (n = 28, including 24/28 meticillin-resistant), enterococci (n = 15), and others (n = 5). Q/D was administered most frequently by central venous infusion, 3 times a day (68/74 patients); the mean and median dose per infusion was 7.4 mg/kg and the mean duration of treatment was 15.6 days. A combined antibiotic therapy was used in 70/74 patients (a glycopeptide in 41/54 staphylococcal infections). RESULTS: A Clinical success at the end of treatment was obtained in 40/74 patients (54%; CI 42.1%-65.7%) [the analysis included 25 patients (34%) with an indeterminate clinical response, categorized as failures] et and 39/73 patients (53%) at the follow-up [including 22 deaths (30%), categorized as failure at the follow-up]. The end-of-treatment success rate in patients with staphylococcal and enterococcal infections was respectively 30/54 (56%) and 8/15 (53%). The safety analysis indicated that 24/74 patients presented at least one treatment-related intercurrent event (possible or probable relationship), the most frequent ones being digestive disordersturbances, signs of venous intolerance, or diffuse or muscular pain. CONCLUSION: Q/D has demonstrated a therapeutic potential in a variety of Gram-positive infections (staphylococcal and enterococcal) in patients with no therapeutic antibiotic alternative, and the type of the intercurrent events reported was consistent with those expected ones in this population of seriously ill patients. PMID- 11899684 TI - [Inappropriate prescription of heparin at curative doses in the hospital. Can the information to prescribing physicians decrease misuse?]. AB - OBJECTIVES: To control whether prescriptions of curative doses of heparin (non fractioned heparins, enoxaparin, tinzaparin) in the hospital complied with the official recommendations; to provide the physicians with information adapted to the recorded misuse and to evaluate the influence of this information. METHODS: A prospective study was conducted between May and October 1999 on the prescriptions of 20 residents from 6 services in 3 phases: phase P1 with initial evaluation (particularly on the indications for heparin, the molecule administered, initial dose and monitoring), phase P2 with analysis and diffusion of an adapted information and phase P3 with final evaluation. RESULTS: 111 inpatients were included in the phase P1 (66.7% aged over 75 years, 18.9% with creatinin clearance below 30 ml/mn) and 101 inpatients were included in the phase P3 (56.4% aged over 75 years, 10.8% with renal failure). During phase P1: among the prescriptions of low molecular weight heparin (LMWH) 54.3% did not comply with the official recommendations; initial doses were too high in 15.3% of patients; mean initial doses of LMWH were not adapted to age, weight or creatinin clearance. Only 58.5% of patients had their platelets monitored. On the other hand, 15.3% of patients exhibited heparin side effects. During phase P3, the main modifications in prescriptions were a reduction in inappropriate indications for LMWH, reduction in LMWH prescriptions in patients aged over 75 or with excessive body weight or with renal failure, and increased platelet monitoring, but without significant difference. On the other hand, mean initial doses of each heparin were not modified. Heparin complications decreased but not significantly. CONCLUSION: This study highlights a real context of heparin prescription at curative doses, often differing from clinical studies, particularly with regards to age, renal failure and comorbidity; prescriptions often unadapted to official recommendations on indications, dose and monitoring; a real but limited influence of appropriate information for the physicians, which partially depends on the accuracy of official recommendations, particularly in patients with increased hemorrhagic risk. PMID- 11899685 TI - [Anorexia nervosa and Crohn disease: diagnostic intricacies and difficulties. 3 cases]. AB - INTRODUCTION: Diagnosis of anorexia nervosa is readily evoked in young girls who associate decreased food intake and weight loss. Among the differential diagnoses of anorexia, it is important to underline the preponderant place occupied by Crohn's disease. OBSERVATIONS: We report three cases of young 18 to 25 year-old girls, initially treated for anorexia nervosa in a psychiatric department. Diagnosis of Crohn's disease was made within 5 to 13 years. The clinical and biological characteristics are reported for each case. COMMENTS: In view of the frequency of digestive disorders concomitant to eating disorders, the distinction between anorexia and digestive disease is particularly delicate. It requires appropriate somatic and biological assessment that, when confronted with clinical and/or biological atypia, may query the initial diagnosis of anorexia. PMID- 11899686 TI - [Endotracheal prosthesis treatment of upper respiratory manifestations of atrophic polychondritis]. PMID- 11899687 TI - [Intrapancreatic lipoma: a rare, easily diagnosed tumor]. PMID- 11899688 TI - [Primary malignant non-Hodgkin's lymphoma of the cervix uteri]. PMID- 11899689 TI - [Lyme borreliosis hepatitis]. PMID- 11899690 TI - [Therapeutic management of primary pulmonary hypertension]. AB - SEVERITY AND TREATMENT: The management of primitive pulmonary hypertension starts by the evaluation of its severity, based on the New York Heart Association's (NYHA) functional classification and right cardiac catheterization data. Hemodynamic criteria of poor prognosis are: an increase in right atrial pressure > 10 mm Hg, a decrease in cardiac output (cardiac index < 2.2 l/min/m2), an increase in pulmonary vascular resistance > to 20 mm Hg/l/min/m2, and saturation of the oxygen content in venous blood < 63%. These prognostic data will condition treatment. CALCIUM CHANNEL BLOCKERS: Are the only vasodilators that have demonstrated efficacy in primitive pulmonary hypertension (PPH). However, only 20% of patients respond to calcium blockers. To determine a positive response to these agents, a carbon monoxide inhalation test is required during right cardiac catheterization. Prescription of calcium channel blockers to non-responders is often responsible for adverse events, from enhanced dyspnoea to sudden death. Continuous intravenous infusion of epoprostenol has considerably improved the prognosis of PPH and related pulmonary hypertension (PH). Despite the constraints, such treatment should be proposed to NYHA functional class III and IV patients not responding to carbon monoxide tests, until new, equally efficient products have been launched on the market. If epoprostenol fails or provokes severe side effects, pulmonary or cardio-pulmonary transplantation should be envisaged. PMID- 11899692 TI - Private surgical facilities must ensure safe nursing care. PMID- 11899691 TI - True health reform, not privatization. PMID- 11899693 TI - The healing of art. PMID- 11899694 TI - [Immunopathology of mycobacterial infection]. AB - In mycobacterial infections, cellular immune response plays a fundamental role in the eradication of this bacteria. However, often it is responsible for perpetuation and severity of tissue damage. On the other hand, the mechanisms employed by mycobacteria to evade immune response explain its chronicity, its difficult eradication and the low efficacy of vaccines. PMID- 11899695 TI - Mycobacterial infection from the cellular point of view. PMID- 11899696 TI - [The role of human genetic factors in susceptibility to tuberculosis]. AB - Tuberculosis has been one of the most important illnesses in the history of the world, but it was never understood why only some people, and not others, develop the disease. It was assumed that human genetic factors play a role in susceptibility, but until the advent of molecular markers, it was never possible to convincingly separate inheritance from the compounding factors of environment and exposure to the bacillus. In recent years particular polymorphisms of several human genes have been shown to be correlated with susceptibility to TB: NRAMP1, Vitamin D receptor, Interferon gamma receptor, IL-12 and its receptor, several HLA haplotypes and there are probably several others that will be discovered. Nevertheless, no single gene appears to play a dominant role in the total TB burden of any population, and exposure of the individual to the bacillus and the environment and nutritional state of the individual also seem to play an important role in determining who will develop the disease. PMID- 11899697 TI - [Cellular cycle control in mycobacteria]. AB - Possibly the failure to eradicate the tuberculosis is linked to the pathogenesis of the causative organism, Mycobacterium tuberculosis. These bacteria can lie quiescent in the human host and later multiplies leading to a higher density of bacteria to increase and, hence at progressive tuberculosis. Particularly unknown is how the mechanism work and which are the molecules responsible for determining if the cells goes to an active replicate process, or in contrast to establish a dormant state. Many genes involved in DNA replication, chromosome segregation and cellular division have been cloned and analysed, and in some bacteria, as E. coli, the cellular cycle events have been described. However, the factors coupling these events are unknown. PMID- 11899698 TI - [The National Program of Tuberculosis]. AB - In this articulate it summarised the objectives and results of the National Program of Tuberculosis during 1999, when the rate of Tuberculosis all the forms were of 26.1 x 100.000 habitants and with results of 80% of cures in the cohort corresponding to 1998, the recommendations also included to improve the execution of the same one. PMID- 11899699 TI - [Bacteriological techniques for the design and identification of Mycobacterium tuberculosis]. AB - This review summarizes the classic bacteriologic techniques used to diagnose M. tuberculosis and others mycobacterial infection. This methodology involves special methods such as staining, recovery in culture media and identification by phenotypic and biochemical characteristics. New methods for rapid diagnosis are been developed and the laboratories should be alert in order to incorporated them in their routine. PMID- 11899700 TI - Molecular diagnostic laboratory design--The Amsterdam experience. PMID- 11899701 TI - [Non conventional diagnostic methods for tuberculosis]. AB - This revision show the advantage and disadvantage of some diagnostic methods for tuberculosis and mycobacterium identification. PMID- 11899703 TI - Resistance to antimicrobial agents in Mycobacterium tuberculosis. AB - According to the World Health Organization (WHO) more people will currently die of tuberculosis (TB) than in any other year in history. Of equal concern are the emergence and nosocomial transmission of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis. Only recently, with the advent of new molecular biological techniques, the mechanisms of drug resistance in TB bacilli are more and more understood. In M. tuberculosis, the primary mechanism of drug resistance seems to be exclusively confined to chromosomal DNA and not, as in other bacteria, to mobile genetic elements as well. PMID- 11899702 TI - The molecular epidemiology of tuberculosis in Caracas, Venezuela, with IS6110 DNA fingerprinting. AB - In this retrospective study we asses the molecular epidemiological situation of Tuberculosis of the city of Caracas, Venezuela in the year 1994, applying IS6110 DNA Fingerprinting of clinical isolates. Fingerprinting of Mycobacterium tuberculosis strains of sixty-four patients TB patients from all the 5 districts of the city revealed fifty-one distinct IS6110 patterns. Isolates from 20 patients (30%) had fingerprints that were shared with at least one other patient. Based on this sampling we conclude that at least a third of the tuberculosis cases in Caracas in the year 1994 were the result of recent and ongoing transmission, indicating micro-epidemics in the town. PMID- 11899704 TI - [Characterization of the expression and function of SigM an ECF sigma factor in mycobacteria]. AB - The survival of M. tuberculosis within the macrophage depends on its ability to respond to oxidative stress, and the ECF subfamily of sigma factors likely play an important role. We studied SigM, a sigma factor whose gene is located near the origin of DNA replication. In both M. smegmatis and M. bovis BCG, the expression of sigM was induced at high temperature and in stationary phase. Mutants of M. smegmatis without an intact sigM were defective for survival in oxidative stress and also for the induction of thioredoxin reductase activity in oxidative stress. The thioredoxin system reduces disulfide bonds that are formed in oxidative stress. SigM thus appears to regulate thioredoxins and forms part of the bacteria's complex protective responses. PMID- 11899705 TI - [Chromosome replication in mycobacteria]. AB - Variation in growth rate has provided a basis for the broad classification of Mycobacterium: (i) the slow-growing class includes the human pathogens Mycobacterium tuberculosis and M. leprae, and (ii) the fast-growing class includes saprophytic nonpathogens, such as M. smegmatis. An intimate association between DNA chromosomal replication and growth rate have been suggested. However, the molecular basis of this relation is unknown. In this article, we summarise our recent work on the study of the origin replication region of some species of mycobacteria, determination of the factors regulating the initiation of DNA replication and the characterisation of the main factor of the replicative machinery, the DnaA protein. PMID- 11899706 TI - [Sliding motility and biofilm formation in mycobacteria]. AB - Using as a model Mycobacterium smegmatis, a non-motile microorganism, we have studied for the first time in mycobacteria the phenomenon known as sliding motility, as well as the process of biofilm formation. A screen of random transposon mutants was performed in order to identify the genes required for mycobacterial sliding over the surface of motility plates. The genetic analysis described here has been published recently. The genes required for sliding and for biofilm formation (mps and tmtpC) are involved in the biosynthesis of the glycopeptidolipids (GPLs) and their transport to the mycobacterial capsule. Based on our results, we suggest a model for the role of the GPLs in both phenomena. PMID- 11899707 TI - [Tuberculosis: pathogenesis and clinical manifestations]. AB - In this revision we present the general aspects of tuberculosis in relationship with its pathogenesis and clinical manifestations. PMID- 11899708 TI - [Transcription regulation in the ribosomal RNA operon of mycobacteria]. AB - Tuberculosis is a extremely important infectious disease, caused by the bacilli Mycobacterium tuberculosis. One of the characteristic of this bacteria is its very slow rate of growth, that allows it to survive for long periods of time inside the host cells. Among the genetic elements involved in growth regulation the operon rrn is of extreme importance. This operon contains the genes that code the three rRNA molecules, essential components of the bacterial ribosome. The tuberculosis bacilli, differently from most of the microorganisms, has a single copy of the rrn operon per genome, meaning that it must be submitted to very strict control mechanisms. Another important conclusion is that the sequences of the rrn operon constitute ideal targets for anti-mycobacterial drugs. In this work we have studied some of the elements involved in transcription control in M. tuberculosis, particularly those present in the leader region of the operon. By using basic molecular biology techniques we have identified sequence elements in the leader region that seem to be involved in the control of transcription elongation, by a mechanism related to anti-termination. PMID- 11899709 TI - [Diagnosis of tuberculosis meningitis by detection of adenosine deaminase activity and amplification of nucleotide sequences with PCR]. AB - Tuberculous meningitis (TBM) is the most severe and lethal form of tuberculosis. The rapid bacteriological diagnosis with the conventional techniques is nearly impossible in TBM. There for many patients are treated with anti-TBC drugs without a definitive diagnosis. A more fast and accurate diagnostic method is necessary, in order to initiate the treatment on time to prevent the irreversible neurologic sequel or death. We evaluated the use of two rapid methods: Adenosine deaminase activity (ADA) and polymerase chain reaction (PCR) for IS6110 and mtp40 sequences on cerebrospinal fluid (CSF) from chronic meningitis patients. For ADA activity > 8.0 U/L the sensibility and specificity was 80% and 91%. PCR sensibility was 80% and specificity 97%. ADA activity and PCR on CSF could be specially useful as complementary tools in the early diagnosis of TBM. PMID- 11899717 TI - [A general approach to the structural shape optimization using genetic algorithms and geometric design elements]. AB - Structural optimization is an engineering field which deal with the improvement of existing solutions or even more find new solutions that are better than the previous ones under some selected criterion. Shape optimization is a research area in this field and it is involved in developing new methodologies to find better structural design based on the shape as resistant element, as for example solutions with the less stress concentration zones and made with the minimum amount of material. The goal of this doctoral dissertation is to present and discuss a general structural shape optimization methodology able to optimize several structural systems or mechanical devices. The approach presented herein is based on global search optimization tools such as Genetic Algorithms and geometric design elements by means of beta-splines curves and surfaces representation. Finally the great versatility of the developed tool is presented and discussed with an application example. PMID- 11899712 TI - [Biomedical images segmentation by the regions growth method]. AB - The study of the different fiber types present in muscle tissue sample images allows the investigators to identify some metabolic and contractile muscle properties, and in general, its physiological state. The image segmentation, which identifies regions or contours of objects within the image, is a fundamental step of the on-line image analysis that can help in this study. However, this kind of images, in general, are very diffuse and with low contrast, so it is difficult to identify the different objects within the image. The main objective of this work is to propose a two-phase method for segmentation of digitized images of cross sections of rat muscle tissue. The first phase allows obtaining an initial group of homogeneous regions by using a uniformity criterion. In this phase we use a division and union of regions algorithm. The purpose of the second phase is to group together in an image the regions corresponding to a given fiber type. The results obtained show an appropriate identification of the regions of interest on the images studied. PMID- 11899718 TI - Who's counting ROI? PMID- 11899719 TI - Taking the lead in regulatory reform efforts. PMID- 11899720 TI - Health care is casualty in current budget battle. PMID- 11899721 TI - Nowak: success depends on commitment to enhancing quality, staff morale. PMID- 11899722 TI - Medical advancements determine health-system capacity requirements. AB - As the rate of advancements in medical technology accelerates, healthcare executives need to anticipate the impact that these advancements will have on health-system demand and capacity requirements. Determining these requirements is critical but difficult. Many hospitals today are operating at capacity, particularly in such areas as the emergency department, operating room, and critical care unit. Key market forces are transforming the way health care is organized, delivered, and financed. One such market force is the escalating pace of medical advancement. The healthcare industry lacks a centralized resource that tracks such developments and assesses the impact on hospital service-line demand and capacity requirements. By following an approach designed to help determine these requirements, Yale-New Haven Hospital, New Haven, Connecticut, was able to forecast its future capacity requirements. PMID- 11899723 TI - A managed care cycle provides contract oversight. AB - In response to poor payment performance by health plans, providers are realizing that managed care contracts require systematic, ongoing management rather than a periodic focus. An effective managed care cycle that encompasses strategy development, implementation of the strategy through contracting and operations, and monitoring of contract performance can accomplish this needed oversight. Each phase requires specialized management tools, skills, and staff. Because of the importance of managed care to the provider's financial viability, a wide range of persons should be involved in the managed care cycle, including the board of directors, business office staff, senior management, and finance staff. As providers embrace a more structured approach to managed care, they will increase their chances of receiving accurate contracted payments. PMID- 11899725 TI - Corporate integrity agreements: making the best of a tough situation. AB - Healthcare providers increasingly are entering into corporate integrity agreements as part of settlements with the Federal government in fraud-and-abuse cases. Providers pursue these settlements to avoid the costs of defending themselves against fraud charges. However, the costs relating to the long-term compliance activity mandated in the settlement's corporate integrity agreement also can be substantial. These costs include significant staff resources that must be devoted to compliance efforts demanded by the agreement and the required engagement of consultants to monitor the organization's compliance. Healthcare financial managers should be familiar with the elements of a typical corporate integrity agreement and understand strategies for negotiating such an agreement. Effective negotiations can help minimize the organization's costs of compliance with the agreement and facilitate its ongoing implementation of the agreement. PMID- 11899724 TI - "Must-know" legal issues for healthcare CFOS. AB - Healthcare financial managers are finding that regulatory agencies' customary focus on Medicare and tax-exemption concerns is shifting to other issues directly related to financial management. Regulatory agencies, such as the Securities and Exchange Commission, are scrutinizing not-for-profit and charitable trust law matters. Because of this renewed attention to existing regulations, healthcare financial managers need to increase their diligence in ensuring their organizations' compliance with those laws governing areas for which they traditionally have been accountable, including auditor relationships, restricted gifts, consulting arrangements, investment management, and financial reporting. PMID- 11899726 TI - Using the OIG model compliance programs to fight fraud. AB - Many healthcare organizations already have implemented compliance programs for their facilities. However, in light of recent fines and continued scrutiny of such programs by the HHS Office of Inspector General (OIG), healthcare organizations should consider reviewing their current programs against the OIG's relevant model compliance program. Although healthcare organizations are not required to adhere strictly to OIG's model programs, they would benefit from ensuring that their programs meet all the OIG's requirements. The common, minimum elements suggested by the OIG model programs include development and distribution of written compliance policies, the designation of a chief compliance officer to manage the program, the development of a corrective action and enforcement system, and the use of audits to monitor compliance. Using these models as guides, healthcare organizations should be better able to avoid the possibility of fraud and abuse within their organizations. PMID- 11899727 TI - Medicare+Choice: what lies ahead? AB - Health plans have continued to exit the Medicare+Choice program in recent years, despite efforts of Congress and the Centers for Medicare and Medicaid Services (CMS) to reform the program. Congress and CMS therefore stand poised to make additional, substantial reforms to the program. CMS has proposed to consolidate its oversight of the program, extend the due date for Medicare+Choice plans to file their adjusted community rate proposals, revise risk-adjustment processes, streamline the marketing review process, enhance quality-improvement requirements, institute results based performance assessment audits, coordinate policy changes to coincide with contracting cycles, expand its fall advertising campaign for the program, provide better employer-based Medicare options for beneficiaries, and take steps to minimize beneficiary costs. Congressional leaders have proposed various legislative remedies to improve the program, including creation of an entirely new pricing structure for the program based on a competitive bidding process. PMID- 11899728 TI - Rising healthcare spending gives pause to Congress and the public. AB - A recent Centers for Medicare and Medicaid Services (CMS) report announced that the nation's overall rate of growth in healthcare spending rose for the second consecutive year in 2000. The increase occurred in both private and public sectors of the healthcare industry, affecting areas such as prescription drugs, hospitals (including both inpatient and outpatient services), and freestanding nursing homes and home health agencies. CMS's findings may signal the beginning of a new period of healthcare inflation, accompanied by calls from Congress and the public for greater cost controls on the part of providers. PMID- 11899729 TI - Achievement and balance: what do managers really want? PMID- 11899730 TI - Biometrics technology adds innovation to healthcare organization security systems. PMID- 11899731 TI - Data trends. CFOs rely on several measures for decision making. PMID- 11899732 TI - [The new Urolan]. PMID- 11899733 TI - [Treatment with shock-wave lithotripsy in children: our experience]. AB - OBJECTIVES: To analyze the efficacy and complication rates of extracorporeal shock wave lithotripsy in the paediatric age group. PATIENTS AND METHODS: From 1989 to 2000, 29 children (mean age 8 years, range 1-14) underwent ESWL for urinary calculi at our institution. A total of forty-four ESWL sessions were performed in 36 calculi and a previous double pigtail stent was inserted in 11 patients. Intravenous anesthesia was used in the vast majority of cases. RESULTS: Complete removal of all stone fragments was achieved in 24 (66.7%) renal units after a first session, 6 (16.7%) after a second session. The rest of the patients became stone-free after 3 sessions in 3 patients, open lithotomy in 1 patient and ureteroscopy in another patient. In one case a watchful waiting was decided after the failure of the first session of ESWL. Mean hospital stay was 3.2 days (range 1-11) for each session. Complications appeared in 10 patients: 4 had renal colic, 3 haematuria, 2 fever and 1 subcutaneous hematoma. CONCLUSIONS: ESWL is a safe and effective treatment for paediatric urolithiasis so it should be considered the first-line treatment. PMID- 11899734 TI - [Xanthogranulomatous pyelonephritis associated with pregnancy, in ex-transplant donor with single kidney]. AB - We present the first case, up to our knowledge, of XGP in pregnant woman, in solitary kidney (related living kidney donor). Therapeutic abortion was not resolutive and nephron sparing surgery (espeleostomy) was successfully performed. Fourteen years later the renal function is still normal and 2/3 of renal parenchima are preserved. Literature review is pointed out. PMID- 11899736 TI - [Primary retroperitoneal tumors: our caseload]. AB - Primary retroperitoneal tumors are a very uncommon group of neoplasias in urology. Sixty-four primary retroperitoneal tumors admitted and treated in our hospital from january 1974 to october 2000 were reviewed retrospectively. Clinical presentation, diagnostic, treatment and evolution are analyzed. Five cases were benign (7.8%) and the remains malign (92.2%). Mesodermic tumors were the most frequent. Surgery was performed in 59 patients (92.2%). Radical resection was possible in 100% of benign tumors and 44.5% of malignant tumors. Palliative radiotherapy was performed as the only treatment in 3 patients. Two patients received only symptomatic treatment. Adjuvant chemotherapy (32 patients, 50%) and radiotherapy (19 patients, 29.6%) completed the treatment. Benign tumors 5-year global survival was 100%, malignant tumors 1-year survival was 47.4%, 3 year survival 15.2% and 5-year survival 10.1%. Mean survival was 20.15 months. As it's frequent to find an advanced neoplasm at the diagnostic, surgery must be planned with radical intention. Radiotherapy and chemotherapy could be useful in the therapeutic strategy of these tumors with poor prognosis. PMID- 11899735 TI - [Prostatic brachytherapy indications and technique]. AB - Prostate cancer is an important health problem, mainly in elderly men. It is the second cause of death among men in USA ant the third at the "Registro del Cancer de Tarragona", behind both the lung and colorectal cancer. About the 58% of the newly diagnosed cancers are localized, therefore, they have to be treated with curative intention. Radical prostatectomy is considered the gold standard treatment for organ confined prostate cancer in our country. On basis to the experience of American groups and the improvement of both, image techniques and dosimetric calculation, brachytherapy has been brought in as a new option in the treatment of localized prostate cancer. We started our program of brachytherapy for prostate cancer on May 2000. We have performed 51 procedures by now. Our protocol and the technique to perform a prostatic brachytherapy are described following. PMID- 11899737 TI - [Leydig cell tumor: report of 8 cases and review of the literature]. AB - Leydig cell tumor is the most frequent non-germ cell tumors of testis, included in the group of specialized gonadal stromal neoplasms. It has a low incidence, accounting for 1-3% of testicular neoplasms. This tumor is characterized by its endocrine manifestations, due to the tumor's capacity to secrete hormones. We report eight cases, including the description of their clinical, diagnosis and therapeutic features, as well as their follow-up. We also make a review of the literature about this rare testicular tumor. PMID- 11899738 TI - [Interpretation of results of clinical trials in benign prostatic hyperplasia]. AB - The random clinical trial (RCT) is the most suitable study to evaluate the treatment effectiveness in the benign prostatic hyperplasia (BPH). Although most of the urologists will not collaborate in a RCT development, they will treat BPH patients, so it is very important to know if a CRT in BPH is well designed and their conclusions are correct. The aim of this article is to give the basic elements of analysis that urologists need in order to evaluate the quality and the level of evidence of a RCT in BPH. This article emphasizes the three main elements of a RCT: to check if the study has been correctly performed (internal validity), to evaluate if the treatment achieves an important clinical improvement (relevance of the results) and the applicability of the results in our patients (external validity). The article shows that to analyse these elements common sense and clinical judgment are needed rather than statistical knowledge. PMID- 11899739 TI - [Apoptosis in renal adenocarcinoma. Expression of bcl-2 in locally confined tumors]. AB - INTRODUCTION AND OBJECTIVE: Bcl-2 is a proto-oncogene known to be a negative regulator of apoptosis, whose expression conferring prolonged cell survival and contributing to tumorigenesis. Inconsistent results concerning bcl-2 expression and the frequency of apoptosis were noted in renal cell carcinoma. To investigate a possible role of bcl-2 protein in renal cell carcinomas, we analyzed its expression and relationship with clinical and pathological parameters, including prognostic impact. METHODS: 58 patients diagnosed of renal cell carcinoma stage pT1, pT2 and pT3a N0 M0 (TNM 1997) were treated by radical or partial nephrectomy. We analyzed clinical and pathological parameters including bcl-2 expression in paraffin-embedded tumor samples using immunohistochemical technique. RESULTS: Bcl-2 immunopositivity was detected in 44/58 of the samples in different grades of intensity. There was no correlation of nuclear grade, tumoral size, stage or recurrency with bcl-2 immunopositivity. Bcl-2 expression was not related to prognosis if we divided all cases into subgroups according of stain intensity. CONCLUSIONS: Bcl-2 expression was not related with any pathological parameters; size, nuclear grade and stage or prognostic. PMID- 11899740 TI - [Urethral amyloidosis]. AB - Localized amyloidosis of the urethra is a rare pathological entity. Biopsy is required to make the appropriate diagnosis. Although localized therapy is available for obstructing, symptomatic lesions, asymptomatic lesions may be followed with conservative management and spontaneous regression has been reported. An appropriate medical evaluation should be performed to determine the presence of systemic amyloidosis. PMID- 11899741 TI - [Unilateral renal cystic disease]. AB - Unilateral and localized cystic disease of the kidney, is characterized by the substitution of either all or a portion of one of the kidneys, by no encapsulated, multiple simple cysts. We present a case--to our knowledge the first in the Spanish bibliography-, of this rare benign condition that needs to be recognized to be differentiated from more transcendent ones. Contrary to autosomal dominant polycystic kidney disease, it is unilateral, it is not familiar, and it is neither progressive nor associated with renal insufficiency or with cysts in other abdominal organs. Lack of encapsulation allows, in general, to distinguish it from multilocular cystic nephroma and cystic renal carcinoma. PMID- 11899742 TI - [Encrusted pyelitis. Lithiasic disease with infectious etiology]. AB - We report on two new cases of encrusted pielitis, a lithiasic disease of infectious ethiology--Corynebacterium of D group-. The clinic diagnostic is difficult and this disease develops in immunosuppressed patients, mainly in renal transplanted ones. One of our two cases is diagnosed in a patient with a transplanted kidney and the other one develops the disease within her native kidneys. We remark on the clinic features and therapeutic options. PMID- 11899743 TI - [Synchronous primary urologic triple neoplasia. Report of a new case and review of the literature]. AB - Herein we present the case of a patient who went to the emergency department complaining of haematuria. With a conservative management, the radiologic and endoscopic studies show a bladder tumour and an incidental left renal mass. After several radical procedures, the last a cystoprostatectomy, it was shown that the patient had a new tumour inside his prostate gland. We comment the surgical procedures, the clinical evolution and the five years follow-up. We review the few cases described in the literature. PMID- 11899744 TI - [Localized sarcomatoid renal carcinoma. Which factors may predict its course?]. AB - Sarcomatoid renal carcinoma constitutes an uncommon variant of renal carcinoma. Typically its behaviour is more aggressive than other forms of renal carcinoma and usually it is diagnosed at advanced stages. The main prognostic factor is the clinical stage at the time of the diagnosis. However, some cases initially are diagnosed at low stages having a poor prognosis, probably because there are other factors than the clinical stage which determinate its outcome. We report a case of sarcomatoid renal carcinoma confined inside the kidney capsule at the time of the diagnosis which had a rapidly bad progression. We review the characteristics of this tumour variant and analyse what factors, basically histological, could be helpful to predict its evolution. PMID- 11899745 TI - [Extramedullary hematopoiesis, a simulator difficult to diagnose]. AB - The extramodular hematopoiesis is the result of the bone marrow sever chronical hypofunction, being the genitourinary affection very unusual. We are presenting a singular case by its own clinicopathology consequences and exceptional incidence in context of a upper tract urothelial tumor. PMID- 11899746 TI - [Duplication of the male urethra]. AB - Duplication of the male urethra is an uncommon congenital malformation. The majority of cases are diagnosed during infancy, and are detected by observing two urethral meatus, or by the appearance of some form of complication, normally of an obstructive nature. Micturitional cystourethrography is an essential test, both in confirming diagnosis of this pathology, and in determining the type of urethral duplication. Excretory urography is also recommended due to its association with other possible urological and extra-urological congenital disorders. Treatment is based on the patient's clinical symptoms, with a waiting period considered advisable for asymptomatic cases. Here we present a case of incomplete urethral duplication, diagnosed from a bladder outlet obstruction, secondary to stenosis of the urethra, which was treated with an endoscopic urethrotomy and distal septotomy, obtaining excellent clinical results. PMID- 11899748 TI - Current treatment options in systemic Sclerosis (Scleroderma). AB - Systemic Sclerosis (SSc) or Scleroderma is a generalized autoimmune disease with variable involvement of the skin and major organs. Etiology and pathogenesis are still largely unknown, but a variety of humoral and cellular autoimmune phenomena can be observed, and a pivotal role of T lymphocytes in SSc pathogenesis is postulated. The rarity of the disease, the wide spectrum of clinical manifestations and severity as well as a variable course render therapy in SSc a major challenge. In view of the immunopathogenesis of SSc, many (presumed) immunomodulatory agents have been used, but no single agent has been proven to be convincingly effective. Trials with extracorporeal therapies (such as photopheresis, plasmapheresis) or even stem cell transplantation are in progress. In contrast to the hitherto unsuccessful therapeutic approaches for the overall disease course, some life-threatening organ manifestations can often be treated successfully, e.g. interstitial pneumonitis with i.v. cyclophosphamide and scleroderma renal crisis with ACE inhibitors and haemodialysis, respectively. Furthermore, pharmacological and supportive treatment of Raynaud's phenomenon and gastrointestinal involvement can alleviate the burden of the disease. Current therapeutic options as well as hitherto investigated immunomodulators are reviewed in this article. PMID- 11899747 TI - Advances in anti-inflammatory therapy. AB - Our goal was to evaluate the state of nonsteroidal anti-inflammatory drugs (NSAIDs) in the treatment of rheumatoid arthritis (RA), before the introduction of the coxibs. The prerequisite for inclusion was the presence of RA (ACR criteria) plus therapy with an NSAID with or without a disease modifying antirheumatic drug (DMARD). A total of 368 consecutive RA patients (81% women) from the outpatient clinic at the Vienna General Hospital were included. Rheumatoid factor was positive in 62%, the patients' mean age was 60 +/- 14 years. The period of observation was 1972-1998. Seventy-seven per cent of the patients had DMARD and NSAID therapy. NSAID therapy was dominated by diclofenac, accounting for 60% of all therapies. Eighteen other substances were applied more rarely. All NSAIDs together were given for 768 patient years (with a mean duration of therapy of 17 years +/- 21 months). Seventy-two per cent of the patients received GI-protective therapy mainly with histamine antagonists and sucralfate while on nonsteroidal therapy. NSAID toxicity mostly affected the GI tract. There was a similar incidence of GI-related adverse events between patients with and patients without GI protection, mainly dyspepsia and nausea. NSAIDs have the potential to cause adverse events in the GI tract. Therapy with histamine antagonists or sucralfate did not reduce the patients' rate of gastrointestinal adverse events. PMID- 11899751 TI - Functional and health status assessment in patients with rheumatoid arthritis. AB - Assessing functional and health status in patients with rheumatoid arthritis (RA) can provide information on the individual's functioning in routine occupations and on the individual's well-being. Data can be obtained by a variety of functional tests and questionnaires. At the Department of Rheumatology at the Vienna University, a specific assessment for outpatients with RA is performed every 3 months. It consists of functional tests, questionnaires, Visual Analog Scales, joint counts, and parameters of disease activity. These data are used to supplement the rheumatologist's decision about medical management of the disease and about further therapeutic strategies. PMID- 11899750 TI - Immunoadsorption therapy (therasorb) in patients with severe lupus erythematosus. AB - Therapeutic removal of immune complexes and antibodies by plasmapheresis has been used in patients with systemic lupus erythematosus (SLE) since 1974. Modern methods of selective adsorption of immunoglobulins from the patient plasma (immunoadsorption, IAS) have been developed; they deserve to be investigated as a tool in the management of difficult cases of SLE. We report our experience in an uncontrolled series of five consecutive SLE patients, in whom cytotoxic immunosuppression was contraindicated or not sufficient to control the disease. Ig-Therasorb columns containing polyclonal sheep antihuman immunoglobulin antibodies were used for IAS for periods of 4 to 54 weeks. In order to prevent rebound autoantibody production, low doses of normal human immunoglobulin were substituted. Improvement in clinical and laboratory signs of disease activity was observed in all patients. In two patients the effect of cyclophosphamide therapy for lupus pneumonitis and lupus-associated thrombopenic purpura was consolidated. In three patients suffering from pancytopenia or lupus vasculitis, the use of cytotoxic substances could be avoided for more than a year. IAS seems to be a safe replacement of conventional plasmapheresis in difficult cases of severe lupus complications. Although controlled studies are lacking, this method may occupy a few important niches as an adjunct in managing immune complex mediated diseases. PMID- 11899752 TI - Treating rheumatoid arthritis with new disease modifying drugs. AB - Rheumatoid arthritis (RA) is a serious illness that can only be controlled by the appropriate use of disease modifying anti-rheumatic drugs (DMARDs). In spite of the successful use of such substances, and of methotrexate in particular, a large number of patients still experience disease progression. Leflunomide and the two anti-TNF agents, infliximab and etanercept, were therefore warmly greeted as very welcome additions to the rheumatologist's armamentarium. These successful newcomers, their strengths and problems are the focus of the present review. PMID- 11899753 TI - [Paraneoplastic rheumatism--musculoskeletal diseases as a first sign of hidden neoplasms]. AB - Malignancy-associated musculoskeletal syndromes can present in a variety of ways which are not distinguishable from idiopathic rheumatic diseases. Furthermore, there are some rare, but typical syndromes with a high association with neoplasms. To perform a quick and exact diagnosis while avoiding useless invasive and expensive diagnostic procedures is a major challenge for the clinician. This article focuses on the clinical features of paraneoplastic musculoskeletal syndromes and theories about the underlying pathogenesis. We try to highlight those clinical and laboratory aspects which could be a clue to hidden malignancies. Paraneoplastic rheumatic syndromes are rare conditions, but timely recognition can save lives. PMID- 11899749 TI - [The effect of Condrosulf (sodium chondroitin sulfate) on proteoglycan synthesis by human osteoarthritic an bovine juvenile articular cartilage chondrocytes--an in vitro study]. AB - BACKGROUND: Sodiumchondroitinsulfate, Condrosulf, is used in osteoarthritis therapy and belongs to the group of symptomatic slow-acting drugs for osteoarthritis. The aim of this study was to investigate the effects of Condrosulf on total proteoglycan synthesis and cell proliferation in human osteoarthritis and healthy juvenile bovine chondrocytes in vitro. METHODS: Chondrocytes were grown as monolayers and stimulated for 7 (human cartilage), or 4, 8 and 12 days (bovine cartilage) with different concentrations of Condrosulf (100 micrograms/ml, 500 micrograms/ml, 1000 micrograms/ml, 2500 micrograms/ml and 5000 micrograms/ml). Proteoglycan synthesis was measured by [35S]Sulfate incorporation. The cell proliferation rate was determined using a [3H]Thymidin assay. The expression of the cartilage markers aggrecan and collagen type II was assessed by Northern blot analysis. RESULTS: We show that the incubation with Condrosulf did not affect proteoglycan synthesis neither in osteoarthritis, nor in healthy chondrocytes under the present culture conditions. Cell proliferation rate was also not increased by Condrosulf stimulation. The results of the Northern blot assays demonstrated a dose-dependent down regulation of aggrecan expression on mRNA level. CONCLUSIONS: These data indicate a lack of direct anabolic effects of Condrosulf on the biosynthetic activity of cultured articular chondrocytes. The well known ease of clinical symptoms, such as pain or swelling under Condrosulf medication may be interpreted by an interaction with pro inflammatory cytokines. PMID- 11899754 TI - T lymphocyte activation--an inside overview. AB - Activated T lymphocytes play an important role in autoimmune disease. The process of T-cell activation is therefore of significant importance in understanding the pathogenesis of many rheumatic diseases. This process can be observed from outside the lymphocyte, but we have also gained increased understanding of many of the intracellular events of T-cell activation. This review tries to draw out the most important receptors, pathways, and transcription factors involved in the process. PMID- 11899755 TI - The nursing management of diarrhoea and constipation before and after the implementation of a bowel management protocol. AB - Intensive care unit (ICU) patients frequently suffer problems associated with both diarrhoea and constipation. Strategies to optimise the management of these conditions need to focus on improving the communication between staff and ensuring effective treatment is implemented. The team involved in this study developed a Bowel Management Protocol (BMP). The effect of this BMP on the documentation of assessment and management of diarrhoea and constipation was evaluated using a quasi-experimental research design. Data were collected via a retrospective audit of medical records. Two groups of patient records were randomly sampled. The records of 60 patients who were admitted to ICU in the 6 months before the introduction of the BMP were accessed together with the records of 60 patients admitted in the 6 months following the introduction of the BMP. Data were collected regarding patient demographics and the assessment and management of bowel function before and after BMP introduction. The results indicated that a BMP improved documentation of the assessment of bowel function. In addition, there was an improvement in the documentation of nursing intervention in the presence of constipation and diarrhoea. These results have to be interpreted with caution because, despite random sampling over two 6 month periods, there were statistically significant differences in age, length of stay, method of feeding and medical diagnosis between the two groups. Further research into the effectiveness of using a BMP is recommended. PMID- 11899756 TI - Nurses and doctors communicating through medication order charts in critical care. AB - The structure and content of written forms of communication dynamically interact with the social and historical conditions underlying critical care nursing activities. One important form of documentation regularly used in the critical care area is the medication order chart. This paper considers the ways in which medication order charts are used to structure interactions among nurses and between nurses and doctors. The critical ethnographic study upon which this paper is based involved a research group of six nurses who worked in one critical care unit. Data collection methods involved professional journalling, participant observation and individual and focus group interviews. Data analysis identified four major issues for consideration: imbalance between medical knowledge and legal authority; the nurse as go-between and medication expert; coaching the doctor; and the self policing nurse. The critical care nurse's role extends beyond the traditional passive activity of medication administration. By exploring the power relations underlying this role, there is greater opportunity for improved nursing relationships and patient care. PMID- 11899757 TI - Pressure sores in intensive care: defining their incidence and associated factors and assessing the utility of two pressure sore risk assessment tools. AB - Patients in intensive care units (ICU) are at high risk of developing pressure sores and the use of pressure sore risk tools has been advocated as a means of identifying patients at risk. A prospective multi-site observational study was conducted to define the incidence of pressure sores, assess two pressure sore risk scales and to define risk factors relevant to intensive care. Patients (n = 534) were assessed for the presence of pressure sores. The Waterlow and Jackson/Cubbin risk scales were completed each day for 314 and 188 of these patients respectively. A total of 75 pressure sores were recorded. Of these, 34 were present on admission. Of the remaining 41, 16 were classified as Grade 1 and 24 as Grade 2 sores. The pressure sore (PS) incidence was 5.2 per cent. Expressed as PS/1000 patient days there were 18.48 pressure sores per 1000 patient days. The ability of the risk scores to predict pressure sores was tested using a Receiver Operating Characteristic (ROC) analysis. The association of risk score with pressure sores was analysed using a survival function (Kaplan Meier) and variables compared using a logrank test (Mantel-Cox). Factors associated with pressure sore occurrence were developed and tested using a survival regression model. Both risk scales were poor predictors of pressure sores (ROC curve area approximately 70 per cent for both). The factors, coma/unresponsiveness/paralysed & sedated and cardiovascular instability were significantly associated with pressure sores with relative risks of 4.2 and 2.5 respectively. Risk increased as a function of time such that the cumulative risk was 50 per cent at 20 days. PMID- 11899758 TI - In-hospital defibrillation--a new technology and a changing role for critical care nurses. PMID- 11899759 TI - Wolff Parkinson White (WPW) syndrome: what the critical care nurse needs to consider when administering antiarrhythmics. AB - This paper discusses the importance of critical care and emergency nurses having an understanding of why pre-existing cardiac disorders can influence antiarrhythmic treatment. The patient with a pre-excitation syndrome is usually managed in a coronary care unit. However, these patients may be admitted to an intensive care unit (ICU) with complications of Wolff Parkinson White (WPW) syndrome; for example post cardiopulmonary arrest or WPW as a co-morbidity. It is common practice in critical care areas for registered nurses to administer antiarrhythmics without a doctor's prescription in life-threatening situations. Therefore, the critical care nurse must have knowledge of the implications of administering standard antiarrhythmic agents if this patient reverts into a tachyarrhythmia. If antiarrhythmics are administered that are contraindicated in patients with WPW syndrome, then there is potential for deleterious effects. This case study highlights the different pharmacological agents for treating tachyarrhythmias in a patient with WPW syndrome. The paper outlines the correct treatment and discusses the deleterious effects of incorrect administration of drugs in WPW syndrome. PMID- 11899760 TI - Reader shares additional resources for astrocytomas. PMID- 11899761 TI - Drug helps breast cancer survivors. Antidepressant reduces severity and frequency of hot flashes. PMID- 11899763 TI - Surgeon General's report highlights the health impact of smoking among women. PMID- 11899762 TI - Updated Internet site provides information to patients with small-cell lung and ovarian cancers. PMID- 11899764 TI - Understanding the perceived need for complementary and alternative nutraceuticals: lifestyle issues. AB - Nutraceuticals are biological therapies used to promote wellness, prevent malignant processes, and control symptoms. The use of complementary and alternative nutraceuticals increased dramatically after passage of the Dietary Supplement and Health Education Act of 1994. Motivations for use of these products include changes in eating patterns, concerns about adequacy of consumer food supply, and interactions with conventional healthcare providers that are perceived to be insensitive, too brief, or uncaring. By becoming knowledgeable about complementary and alternative nutraceuticals and the nutritional needs of people with cancer, communicating with empathy and patience, and involving dietitians, pharmacist, and other professional providers as needed, oncology nurses can provide accurate information and support for people with cancer and their families. PMID- 11899765 TI - Chemosensitivity testing. AB - Determining which chemotherapy treatment to administer to a patient with cancer is challenging because each patient's cancer is unique. Hundreds of different forms of cancer exist, and tumors of the same type or classification can have very different responses to the same, often standard, treatment. A scientifically based analysis of how the chemotherapy options for an individual patient may affect that patient's specific tumor could benefit both the clinician and the patient. Chemosensitivity testing can provide this information. A Chemosensitivity assay is a laboratory test that is performed by isolating the cancer cells from a tumor specimen, exposing the cells to the desired chemotherapy agents, and evaluating the effectiveness of those agents. The results provide patient-specific tumor information. PMID- 11899766 TI - Molecular cytogenetics and gene analysis: implications for oncology nurses. AB - Current advances in genetics have provided a better understanding of many diseases, including cancer, and will have an impact on oncology clinical practice in an unprecedented way. The molecular cytogenetic techniques of fluorescent in situ hybridization (FISH), comparative genomic hybridization (CGH), and spectral karyotyping (SKY) are providing tremendous insights into genetic information related to cancer by specifically illustrating chromosomal abnormalities that can occur in a patient's cancer cells. The application of these techniques allows for the development of molecular diagnostic tests may be applied to clinical material, which may help to improve the diagnosis and staging of a patient's tumor, particularly in small, premalignant lesions that often are equivocal and difficult to assess. An understanding of these genetic changes will provide a foundation of knowledge for oncology nurses that will lead to significantly improved detection methods, therapies, and disease prevention. As members of the healthcare team, oncology nurses must be knowledgeable about the rapid expansion of genetic information. Oncology nurses are in a unique position to translate this information to patients and their families and, ultimately, enhance comprehensive care through patient education and advocacy. PMID- 11899767 TI - Chronic myelogenous leukemia: an overview. AB - Important advances have been made in the treatment of chronic myelogenous leukemia (CML). In the past two decades, treatment of this disease has changed and now includes the use of hydroxyurea, interferon, tyrosine kinase inhibitors, and high-dose chemotherapy followed by hematopoietic stem cell transplantation. Because several relatively effective forms of therapy now exist, the care and management of patients with CML has become more complex. Through continued research and drug development, additional therapies are expected to emerge offering new hope and expanded treatment options for patients with CML. PMID- 11899768 TI - Music, cancer, and immunity. PMID- 11899769 TI - Carcinogenesis: application to clinical practice. AB - The evolution of cancer is an intricate, multistage process involving the interplay of numerous variables that both inhibit and enhance the growth and spread of disease. An understanding of the process of carcinogenesis is vital to successful care of patients with cancer. Nurses play a key role in translating information to patients about prevention and health promotion, diagnostic testing, treatment approaches and side effects, and the need for frequent and long-term follow-up. Many patients and their families are well aware of the continual, often exciting, advances in prevention, detection, and treatment of malignancies. Understandably, they look to oncology nurses and other healthcare professionals for guidance and understanding of how these changes might affect their healthcare decisions. PMID- 11899770 TI - Aplastic anemia. PMID- 11899771 TI - Painful blistered hands and feet. AB - CAE is a self-limiting toxicity seen with several types of high-dose chemotherapy. Treatment of these patients requires pain management and supportive therapy, including wound care that promotes healing, comfort, mobility, and quality of life and prevents infection. Oncology nurses play an important role in monitoring patients for CAE and providing supportive care. PMID- 11899772 TI - A serious look at the undertreatment of pain: Part 1. PMID- 11899773 TI - Arsenic trioxide. PMID- 11899775 TI - Herbs 101. PMID- 11899774 TI - Commonly used herbs: implications for clinical practice. PMID- 11899776 TI - Brachytherapy for head and neck cancer: a case study. AB - Brachytherapy currently is being used as a treatment modality for head and neck cancer. A case study is presented to illustrate the treatment and safety procedures required when brachytherapy is delivered. A multidisciplinary approach is essential to ensure that family and staff exposure to radiation is minimized. Nursing care considerations include preventing airway impairment, maintaining hydration, developing alternative communication methods, pain management, and bowel preparation. Through intensive patient, family, and staff education, patients with head and neck cancer can be treated with brachytherapy safely and effectively. PMID- 11899777 TI - Nursing management of patients receiving brachytherapy for gynecologic malignancies. AB - Brachytherapy is a form of radiation therapy used to treat a variety of cancers. Patients undergoing gynecological brachytherapy face a variety of physical, emotional, and psychosocial issues. Nurses caring for patients receiving high dose rate or low-dose rate brachytherapy must be knowledgeable about radiation therapy principles and technology and the acute and chronic side effects that patients may experience. PMID- 11899778 TI - Campath-1H. AB - Chronic lymphocytic leukemia is a malignant hemato-logical disorder characterzed by an accumulation of immunologically incompetent lymphocytes. The majority of cases (95%) are neoplasms of B-lymphocytes and 2%-5% are T-lymphocytes lymphoproliferative disorders Campath-1H (llex Pharmaceuticals, San Antonio, TX) is a humanized monoclonal antibody that is directed against CD52, a cell-surface antigen present on the lymphocytes of almost all patients with B and T cell lymphocytic leukemia. Campath-1H has shown promising results in early clinical studies, and aproval from the U.S. Food and Drug Administration for the use of this monoclonal antibody in the treatment of chronic lymphocytic leukemia may occur this year. PMID- 11899780 TI - Sepsis and septic shock. PMID- 11899779 TI - My experience with the Cancer Survival Toolbox. AB - Nurses are instrumental in educating patients and their families about how to navigate the cancer experience. Patients often state that they would not know much about their disease and treatment if not for their nurse's teaching. The Cancer Survival Toolbox, a comprehensive set of audiotapes, is a free resource that helps people dealing with cancer develop and use key coping skills- communicating, finding information, making decisions, solving problems, negotiating, and standing up for their rights. Supplemental modules deal with issues facing the underinsured and uninsured, barriers to care for older cancer survivors, and concerns of caregivers. The Cancer Survival Toolbox Group Facilitator's Training Manual is a valuable resource for anyone working with cancer support groups. All healthcare professionals, cancer survivors, and lay caregivers can use the toolbox in a variety of ways. This article offers suggestions about how to ensure that cancer survivors and caregivers have the opportunity to use this unique resource. PMID- 11899781 TI - Rituximab. PMID- 11899782 TI - Gemcitabine hydrochloride. PMID- 11899783 TI - A review of vinorelbine in the treatment of breast cancer. AB - As combinations and sequences of anthracyclines and taxanes increasingly become standard adjuvant treatment for early breast cancer, a major need for new treatment options for metastatic breast cancer will arise. Vinorelbine is highly active in the treatment of metastatic breast cancer, both as a single agent and in combination regimens. Furthermore, it is well tolerated, with a low incidence of subjective toxicities. It is anticipated, therefore, that vinorelbine will become increasingly utilized for treating metastatic breast cancer due to its favorable safety profile, good tolerability, and promising results in combination with other chemotherapy agents. Combinations with trastuzumab and newer molecular targeting agents are being explored. Doublets or triplets of vinorelbine with drugs other than anthracyclines and taxanes could be considered in the next generation of adjuvant and neoadjuvant trials, where it is anticipated that anthracycline/taxane combinations are likely to replace anthracycline/cyclophosphamide combinations as the mainstay of adjuvant treatment. PMID- 11899784 TI - HER2/neu as a predictive factor in breast cancer. AB - One of the current trends in breast cancer research is to identify markers that can predict response to specific anticancer therapies; intense laboratory research and therapeutic trials are exploiting this strategy. The combination of cytotoxic drugs directed at the tumor population with the highest probability of being sensitive to them with molecules targeted at intracellular signaling and cell cycle control pathways, which may be deregulated as part of the malignant process, represents our best hope for improved survival in both early and advanced disease. The transmembrane tyrosine kinase receptor, HER2/neu, has been the subject of much investigation with respect to its prognostic value, predictive value, and as a target of antibody-mediated therapy. Retrospective evidence strongly suggests that HER2 overexpression is associated with decreased disease-free and overall survival in node-positive, and possibly also node negative, breast cancer. Prospective trials have demonstrated that antibodies to HER2 can produce tumor responses in women with advanced disease that overexpresses this molecule. Moreover, the combination of such antibodies with cytotoxic drugs has been one of the few recent strategies to improve survival duration in metastatic breast cancer. The evidence supporting the role of HER2 as a factor predictive of response to hormone therapy and cytotoxic drugs is more ambiguous and requires prospective assessment. The available literature is reviewed herein, with a focus on the predictive value of HER2, potential mechanisms of resistance and sensitivity to various drugs, and future research directions involving this important molecule. PMID- 11899786 TI - The impact of alternative practices on the cost and quality of mammographic screening in the United States. AB - The decentralized structure of health care in the Unites States hinders population-based analysis of breast cancer screening. Our objectives are to model mammography in the United States as a whole, to identify the variables that most profoundly affect cost and efficacy, and to develop a strategy to improve mammography screening from a population perspective. A spreadsheet model was used to represent the variables of mammography screening in the United States. The population-based national screening program in Sweden provides a framework for comparison. The outcome measures are the aggregate cost and the number of cancers detected by mammography. We used deterministic sensitivity analysis to calculate the impact of variation in practice. Aggregate costs of screening in the United States are in the range of $3-$5 billion dollars. The percentage of women screened, cost per mammogram, cancer to biopsy ratio, recall rate, and cost of recall have the most profound effect on the quality and cost of a national screening program. Variance of these high-impact variables, based on the U.S. population, modifies the aggregate cost of screening by over $2 billion. As mammography screening in the United States increases to include all women over age 40, high-impact variables should be optimized to decrease costs and improve breast cancer detection. Our model establishes which parameters are most important. PMID- 11899785 TI - Meta-analysis of adjuvant cyclophosphamide/methotrexate/5-fluorouracil chemotherapy in postmenopausal women with estrogen receptor-positive, node positive breast cancer. AB - Conflicting results have been published regarding the efficacy of adjuvant cyclophosphamide/methotrexate/5-fluorouracil (CMF)-type chemotherapy in postmenopausal, estrogen receptor (ER)-positive women. The Oxford overview suggests real but limited benefit of any chemotherapy in this group of patients but avoids analyzing smaller subsets. We wished to better quantitate the benefit of adding CMF to tamoxifen in postmenopausal ER-positive women with tumor involvement of axillary lymph nodes. Six randomized studies comparing CMF plus tamoxifen to tomoxifen alone in postmenopausal, ER-positive, node-positive women have been published since 1992. They include 2368 patients. We performed a meta analysis of 6 endpoints: survival, disease-free survival, locoregional recurrence, distant recurrence, contralateral breast recurrence, and thromboembolic complications. There was a statistically significant increase in disease-free survival from the addition of CMF-type chemotherapy to tamoxifen in this population; the absolute risk of relapse was reduced by 5.5% at 5 years. Effects of locoregional recurrence were greater than those on overall recurrence. No significant survival benefit was observed. PMID- 11899787 TI - Male inflammatory breast cancer. AB - A case of a 48-year-old male with an inflammatory breast cancer is used to illustrate this uncommon malignancy. The physical examination of thickening and erythema made the clinical diagnosis. Mammographic findings of increased density in the right breast with coarsened stroma and an underlying mass confirmed the clinical findings. The sonographic evaluation revealed a 2-cm ill-defined hypoechoic mass. The pathologic examination of the mastectomy specimen showed an infiltrating duct cell carcinoma with lobular features. Male breast cancer afflicts 1500 men each year. Clinically it must be differentiated from gynecomastia, a much more common and benign condition. PMID- 11899788 TI - Overexpression of plasminogen activator in male breast cancer. PMID- 11899789 TI - All therapy is targeted therapy: the future of systemic therapy. PMID- 11899790 TI - Workers displaced from employment, 1997-1999: implications for employee benefits and income security. PMID- 11899791 TI - New health care marketplace demands much more from governance. PMID- 11899792 TI - Technology is key to reducing medication errors. PMID- 11899793 TI - The business of health care: what level of performance do we need to survive? PMID- 11899794 TI - [Efficiency treatment of new cases of destructive pulmonary tuberculosis in the presence of concurrent iron deficiency anemia]. AB - A hundred and twenty nine new cases of destructive pulmonary tuberculosis were examined. Of them, 95 patients were diagnosed as having first-degree iron deficiency anemia. Anemia was found to have a negative impact on the clinical efficiency of therapeutical measures. For correction of anemia, the use of antioxidants (tocopherol acetate, ascorbic acid) and an antihypoxant (riboxine) yielded better clinical results than did an iron-containing agent (ferroplex). These results were mostly close to those obtained in a group of patients without anemia. PMID- 11899795 TI - [Central hemodynamics in patients with acutely progressive and common forms of pulmonary tuberculosis]. PMID- 11899796 TI - [Multiorgan tuberculosis: central peripheral hemodynamic features]. AB - The paper deals with the study of central and peripheral hemodynamic features in patients with multiorgan tuberculosis. The study is based on the analysis of examination findings and treatment outcomes in 42 patients with concurrent tuberculosis of the lung and vertebral column. A control group consisted of 46 patents with spinal tuberculosis. The authors provide evidence that concurrent pulmonary and spinal tuberculosis substantially makes basic central and peripheral hemodynamic parameters worse in patients with tuberculosis spondylosis, aggravates their general condition, requires cardiotropic therapy on an individual basis, substantially prolongs preoperative preparation and postoperative rehabilitation. PMID- 11899797 TI - [Serum concentrations of trace elements and microcirculation during laser therapy for pneumonia]. AB - The impact of laser therapy in the multimodality treatment for pneumonia on the serum levels of trace elements and microcirculation was studied. A total of 105 patients with pneumonia were examined. The patients were divided into 2 groups: 1) 68 patients received laser therapy and 2) 37 patients took drug therapy alone. There was a more substantial reduction in the permeability of cellular membranes, a significant increase in the serum levels of iron and chromium, better circulation at the expense of a vascular component. The above changes highly correlated with laboratory data and external respiratory function parameters. PMID- 11899799 TI - [Adverse effects in children receiving short-term chemotherapy for intrathoracic tuberculosis]. AB - The incidence and pattern of adverse effects were analyzed in children receiving short-term chemotherapy for intrathoracic tuberculosis in relation to the course of a tuberculous process (complicated and uncomplicated). During chemotherapy, side effects of antituberculous agents were noted in 82 children of 255 examinees. The incidence of side effects was affected by concurrent diseases detected in 52.4% of children showing a poor tolerance, receiving antituberculosis therapy for up to 6 months. Adverse effects were found in 18.2% of children, and in 55.3% with over 12-month chemotherapy. Short-term chemotherapy in children with the uncomplicated course of tuberculosis significantly reduced the incidence of adverse responses as compared to long-term chemotherapy (20 versus 36%) by reducing allergic reactions by 3 times (6.7 versus 19.7%). In children with complicated tuberculosis, the use of 4 drugs in the intensive phase of chemotherapy failed to increase the incidence of side effects (35.5 versus 40.8%). In most children, adverse reactions are reversible and a complete intolerance of antituberculous agents (streptomycin and rifampicin) was observed only in 4% of cases. PMID- 11899798 TI - [New technologies in the surgical treatment of tuberculous spondylitis]. AB - The paper deals with surgical treatment in 452 patients with tuberculous spondylitis. The process involved the cervical, thoracic, thoracolumbar, and lumbar parts in 40 (8.8%), 185 (41.0%), 75 (16.8%), and 11 (2.5%) cases, respectively. The active process was found in 125 (27.6%) patients, its loss was detected in 131 (29.1%) cases; the process abated in 196 (43.3%) patients. There were its complicated forms in 341 (75.4%) patients, para- and prevertebral abscesses in 183 (40.4%), epidural abscesses in 26 (5.6%), pareses and paralyses in 71 (15.7%), fistulas in 38 (8.4%), and membranous symptoms in 23 (5.1%). The diagnosis of tuberculous spondylitis was established up to month 3, in 109 (24.1%) patients, up to month 6 in 208 (46.0%), at months 9-12 in 74 (16.4%), at month 12-18 in 61 (13.5%). The concomitant tuberculous involvement of the lung, pleura, kidneys, joints, lymph nodes, and genitals was observed in 139 (30.7%) patients. The resistance of Mycobacteria tuberculosis to antituberculous drugs (streptomycin, isoniazid, and rifampicin) was detected in 139 (30.7%) patients. Of them, 40 (28.8%) patients were unresponsive to two drugs or more. This group of patients received a combined therapy with lomefloxacin and mycobutin on an individual basis. Surgery was made in all the patients; posterior approaches were applied to 44 (9.7%) patients, in 368 (81.4%) anterior approaches were applied to extrafocal apparatus fixation in combination with autograft corporadesis (n = 70), in combination of an autograft and a biocompatible implant with benemycin application (n = 298). The use of these new technologies may expand the scope of operability, reduce surgical traumaticity, initiate early (at day 7-12) rehabilitation of patients, achieve good and satisfactory clinical and functional results by months 4-8 in 81.1% of cases. PMID- 11899801 TI - [Methods for the control of quality in mass tuberculin tests]. AB - It is possible to achieve a qualitative annual intradermal testing of children and adolescents by the Mantoux test with 2 TE PPD-L and to form Dispensary Registration Group 6 completely only when there is a combination of a mass tuberculin diagnosis, team's well organized work and systematic control methods for its work quality. Standards have been developed for the quality of registration of Group 6 patients and for that of registration of positive Mantoux tests with 2 TE PPD-L, for the completeness of formation of the above group of patients, for organization of controlled chemoprophylaxis in children's collective bodies. PMID- 11899800 TI - [Experience with rovamycin and roxithromycin in patients with chronic bronchitis in the presence of posttuberculous changes]. AB - Rovamycin and roxithromycin were comparatively studied for their effects in 25 patients with chronic bronchitis and posttuberculous changes in the lung. Ten patients received oral rovamycin, 3,000,000 IU, twice daily and 15 had oral roxithromycin, 150 mg, twice daily. The therapy averaged 9 days. Clinical cure was observed in 90% of cases on rovamycin and in 86.7% on roxithromycin. There were no adverse effects of these drugs. The findings suggest that due to their high therapeutical and bacteriological activities, both rovamycin and roxithromycin show their good clinical efficacy in the treatment of exacerbations of chronic bronchitis in patients with residual posttuberculous changes. PMID- 11899802 TI - [Anti-tuberculous IgE antibodies. I. Immunodominant antigens]. AB - It is widely accepted that protection against tuberculosis is provided by the formation of type 1 immune response, which is characterized by the production of IFN-gamma and IL-2. However, type 2 antimycobacterial immune response is also present: specific IgE antibodies that are IL-4 dependent, are usually found in tuberculosis patients. There is elevated production of type 2 cytokines in some cases. Thus, both types of an immune response can simultaneously develop, probably counteracting with each other. It is unknown which of mycobacterial antigens are capable of inducing a preferential type 2 response. To detect these antigens, the authors studied tuberculosis IgE antibodies in the sera of 500 tuberculosis patients by using the ELISA assay with ultrasonic disintegrated M. Tuberculosis H37Rv (sonicate). Antigens recognized by IgE antibodies were found to be localized in the cell wall of mycobacteria. The IgE-response was specific since the sera did not react with the antigens of atypical mycobacteria and other bacterial species. PMID- 11899803 TI - [Red blood cell metabolic changes in patients with acutely progressive pulmonary tuberculosis]. AB - In 52 patients with acutely progressive pulmonary tuberculosis (APPT), their red blood cells were used to determine the values of their energy metabolism by lactate dehydrogenase (LDG) and glucose-6-phosphate dehydrogenase (G-6-PDG), the antioxidative defense from the activity of superoxide dismutase (SOD) and catalase and from the levels of malonic dialdehyde (MDA), as well as the intraerythrocytic level of platelet activation factor (PAF) and 2,3 diphosphoglycerinic acid (2,3-DPGA). The concentration of plasma hemoglobin (Hb) was measured as an indicator of spontaneous hemolysis. In the red blood cells from patients with APPT, the activity of LDG and SOD was found to be drastically suppressed, the content of MDA was increased, the intracellular level of PAF was lowered and the concentration of 2,3-DPGA was very moderately elevated. The plasma concentration of free Hb is thrice as high as the normal value. Following 3 months of chemotherapy with 4-5 drugs, the activity of LDG became higher, yet remaining less than the normal values and that of G-6-PDG significantly dropped. The activity of SOD and catalase increased, but the level of MDA remained high. There was a drastic fall in the intracellular levels of PAF. The plasma concentration of free Hb dropped, but remaining significantly higher than the normal values. Possible ways of correcting the changes found are discussed in the paper. PMID- 11899804 TI - [Activity of pleural fluid adenosine deaminase in tuberculous pleurisy]. AB - Examining the activity of adenosine deaminase in the pleural fluids of 69 patients with tuberculous pleurisy of various etiology from the clinics of Armenia indicated that it was greater than the threshold value of 20 U/L in 95.7 of 47 patients with tuberculous pleurisy. The specificity of this parameter for this disease was 0.91. The prognostic value of the test with positive and negative results was 0.96 and 0.94, respectively. The diagnostic value of the ADA test was 0.94. PMID- 11899805 TI - [Tobacco smoke condensate-induced structural changes in Mycobacteria tuberculosis]. AB - The paper shows how a tobacco smoke condensate affects the anatomy of Mycobacteria tuberculosis. Gas chromatography was used to detect quantitative changes in the composition of fatty acids. Electron microscopy indicated larger microcolonies, thickened lipid capsular cover, increased number of polysomes in experimental mycobacterial strains. PMID- 11899806 TI - [Drug resistance in Mycobacteria tuberculosis in the Barents region of Russia and Norway]. AB - The paper considers drug resistance in Mycobacterium tuberculosis (MBT) in the Barents region of Russia and Norway. Along with a rise in tuberculosis morbidity, there has been recently an increase in the number of patients isolating MBT that are resistant to antituberculous agents. The most dangerous trend is an increase in MDR of MBT strains. In the north-west of Russia there are high MBT drug resistance rated as compared to Norway. There are differences in the determination of drug resistance by the absolute concentration method and by the BACTEC method. PMID- 11899807 TI - [Spontaneous hemopneumothorax, one more possible cause]. PMID- 11899808 TI - [A case of pulmonary histiocytosis X]. PMID- 11899809 TI - [Tuberculosis: problems and research in the world's countries]. PMID- 11899810 TI - [History of the Department of Phthisiatry and Pulmonology of the Kazan State Medical Academy]. PMID- 11899811 TI - [Indication for the surgery in patients with pulmonary tuberculosis]. PMID- 11899812 TI - [The use of global internet in a phthysio-pulmonologist's practice]. PMID- 11899813 TI - [Biopsychological model of chronic pulmonary tuberculosis]. AB - Examining 253 patients with fibrocavernous pulmonary tuberculosis (152 males and 101 females) yielded two variants of a biological psychosocial model of chronic pulmonary tuberculosis for males and females. The key points of the biological constituent are similar in males and females and the psychological and social constituents differ. Patients should receive all data on the disease, its course and prognosis from their physician. The efficiency of sociomedical rehabilitation of a patient with chronic pulmonary tuberculosis largely depends on his/her physician's awareness of a sick patient's inner life. Psychosocial correction should be made purposefully. PMID- 11899814 TI - [Prevalence of bronchial asthma in Uzbekistan]. AB - The prevalence of bronchial asthma has tended to increase in Uzbekistan. It is highest in the Republic of Karakalpakstan and least in the Samarkand Region. Its prevalence differs in some areas of Uzbekistan--highest in children in Karakalpakstan and in elderly persons in Tashkent. The prevalence of bronchial asthma is associated with smoking. PMID- 11899815 TI - [Gastric mucosa as a target in duodenogastric reflux: assessment of lesions with the system of indices]. AB - AIM: To characterize gastric mucosa lesion in patients with duodenogastric reflux (DGR) using an original system of indices evaluating general pathological processes in gastric mucosa. MATERIAL AND METHODS: The trial enrolled 109 patients with chronic gastritis (CG) in combination with opisthorchiasis. These were divided into 2 groups: CG patients with DGR (group 1, n = 58), CG patients free of DGR (group 2, n = 51). Thirty patients with chronic acalculous cholecystitis served control. All the patients have undergone fibrogastroduodenoscopy with biopsy from the antral part of the stomach and its body. To detail morphological picture of the gastric mucosa a number of additional indices were used characterizing exudative inflammation, productive inflammation, atrophic, sclerotic, immunopathological, dysregeneratory processes. RESULTS: Semiquantitative analysis of gastric mucosa in chronic opisthorchiasis patients with DGR by the above indices showed a significant prevalence of exudative (p < 0.05) and productive (p < 0.01) inflammations in the antrum over those in the gastric body. Comparison of the indices in the patients with and without DGR revealed significant differences in activity of exudative inflammation (p < 0.05), dysregeneratory (p < 0.05) and atrophic (p < 0.001) processes in gastric antrum and dysregeneratory and atrophic processes in the gastric body (p < 0.01). CONCLUSION: The indices contribute to more precise assessment of structural shifts occurring in gastric mucosa which is the target for DGR. PMID- 11899817 TI - [Morphological validation of peloidobalneotherapy intensification in patients with duodenal ulcer]. AB - AIM: To study effectiveness of short courses of intensive peloidobalneotherapy in duodenal ulcer (DU). MATERIAL AND METHODS: Esophagogastroduodenoscopy and biopsy before and after the treatment course were made in 124 DU patients. Depending on timing of moor applications, pine and pearl baths, the patients were divided into 2 groups. Patients of group 1 received procedures daily with interval 4-5 hours (intensive course), patients of group 2 were treated each other day. RESULTS: 3-4 weeks of the treatment produced a response in both groups but in group 2 there were no signs of active inflammation in gastric and duodenum mucosa. 2 years later, patients of group 1 had histological evidence of chronic inflammation exacerbation. The latter was absent in group 2. CONCLUSION: An intensive course of peloidobalneotherapy results in attenuation of chronic gastroduodenal inflammation in DU patients. PMID- 11899816 TI - [Characteristics of aggressive and protective factors in erosive lesions of gastroduodenal mucosa]. AB - AIM: To investigate aggressive-protective factors in patients with erosive lesions of gastroduodenal mucosa. MATERIAL AND METHODS: Intragastric pH-metry, tests for Helicobacter pylori, lysozyme activity, content of bile acid in gastric juice, components of gastric mucous gel were made in 106 patients. RESULTS: It is shown that there are two alternative paths of erosions development in the gastroduodenal zone: infectious and non-infectious. CONCLUSION: In patients infected with Helicobacter pylori, lowering of a protective ability of the mucous gel is caused by prevalence of synthesis of immature mucous components while in non-infected patients this happens as a result of high catabolism of mucous components. PMID- 11899818 TI - [Characteristics of non-Helicobacter pylori ulcer disease]. AB - AIM: To characterize clinicomorphological and psychosomatic manifestations of ulcer disease (UD) with reference to Helicobacter pylori (HP) contamination of gastric mucosa. MATERIAL AND METHODS: Out of 411 UD patients HP invasion was diagnosed in 338. RESULTS: Patients with UD having no HP in gastric mucosa showed clinical peculiarities related to changes in psychological and vegetative status. CONCLUSION: The addition of psychopharmacological drugs to conventional antiulcer therapy promoted induction of clinicoendoscopic remission of UD within optimal time and improved quality of life in patients without HP infection in the stomach. PMID- 11899819 TI - [Diagnosis of gluten enteropathy and efficiency of its treatment]. AB - AIM: To develop screening diagnosis of gluten enteropathy (GEP), indications to administration of glucocorticoid hormones and objective criteria of effective treatment. MATERIAL AND METHODS: Clinical, immunological (antibodies-Abs to alpha gliadin, reticulin and endomisium) examinations, enterobiopsy with morpho- and stereometry of small intestinal mucosa were made in 200 GEP patients. The examination was repeated 6 months to 5 years and later after the discharge from the hospital. RESULTS: Mean values of Abs to alpha-gliadin was 6 times higher than normal values. Positive titers of Abs to endomisium and reticulin were in 100 and 87.5% patients, respectively. Formed stool was registered 1.5 times more frequently, polyfecalia occurred 2.5 times less frequently, hypovitaminosis and trophic disorders were relieved 3 times more frequently, malabsorption syndrome reduced in patients given prednisolone vs those untreated with it. CONCLUSION: Screening diagnosis of GEP may be based on Abs tests to alpha-gliadin, reticulin and endomisium. Improvement of clinical condition of GEP patients can be stated by decreased diarrhea, polyfecalia and malabsorption symptoms. The treatment efficacy may be judged by clinical improvement, recovery of morphological structure of small intestinal mucosa, normalization of concentration of Abs to alpha-gliadin, reticulin, endomisium. Administration of prednisolone provides more complete and rapid rehabilitation of the patients. PMID- 11899820 TI - [Interferon status of patients with non-specific ulcerous colitis and its correction with interferon inducers]. AB - AIM: To examine validity of using interferon inductors (amixin and cycloferon) in combined treatment of patients with non-specific ulcerous colitis (NUC). MATERIAL AND METHODS: 113 NUC patients received basic antiinflammatory therapy (glucocorticoids and preparations of 5-aminosalycylic acid). Patients of groups 1 and 2 received interferon inductors (amixin and cycloferon in tables), respectively. Patients of groups 1a and 2a received placebo. All the patients were examined clinically with evaluation of interferon status and cellular immunity before and after the treatment. Colon mucosa biopsies obtained endoscopically were studied histologically. RESULTS: Alpha- and gamma-interferon production by leukocytes was substantially suppressed in all the examinees against normal levels of serum and spontaneous interferon. The interferon inductors stimulated relief of clinical symptoms and eliminated endoscopic signs of the disease. Amixin and cycloferon normalized interferon status in 36.3 and 33.3% of the treated patients, respectively. No response was registered in 9.1 and 8.4%, respectively. Improvement was seen in 54.6 and 58.3%, respectively. CONCLUSION: Cycloferon and amixin, interferon inductors, are effective in the treatment of NUC. PMID- 11899821 TI - [Clinical, biochemical, and morphological characteristics of chronic hepatitis with various etiology]. AB - AIM: To investigate clinicomorphological features in patients with chronic diffuse lesions of the liver with consideration of the etiological factor. MATERIAL AND METHODS: Clinical, laboratory, serological and pathohistological examinations were performed in 67 patients with chronic hepatitis (CH). RESULTS: Out of 48 patients with chronic viral hepatitis (CVH), 32(66.6%) demonstrated association of hepatic lesion with alcohol or drug abuse. Knodell activity in the liver was prevalent in drug abusers with CVH. Fat dystrophy and high de Ritis index were typical for alcoholics. CONCLUSION: Viral hepatic lesions associated neither with alcohol nor drug abuse occur only in 1/3 of patients with CH. PMID- 11899823 TI - [Syndrome of cross-over between primary biliary cirrhosis and autoimmune hepatitis]. PMID- 11899822 TI - [Cytokine system in patients with chronic hepatitis C during treatment with interferon-alfa]. AB - AIM: To study changes in serum levels of interleukine-1 beta (IL-1b), IL-6, TNF alpha (TNFa), HM-HMF and TFR-1 beta (TFR-1b), expression of surface antigens CD14 and CD95 on blood monocytes from patients with chronic hepatitis C (CHC) treated with interferon-alpha (INFa). MATERIAL AND METHODS: Examinations covered 25 CHC patients and 25 healthy controls. Concentrations of proinflammatory cytokines and growth factors in blood serum were measured with ELISA (kits by "R&D systems", USA). CD14 and CD95 antigen expression on monocytes of venous blood were studied using flow cytoflowmeter (Partes, USA) before and after a 12-week course of INFa. RESULTS: Before INFa treatment CHC patients had significantly elevated serum concentrations of TNFa, HM-KSF and TFR-1b. Coexpression of antigens CD14+ and CD95+ was found on 61% of blood monocytes. Three-month INFa treatment lowered levels of TNFa, GM-KSF and CD95+ expression on monocytes as well as TFR-1b concentration in the serum which correlated with a positive trend in the standard clinicolaboratory and virusological indices in the examinees. CONCLUSION: Changes in serum indices of proinflammatory cytokines and growth factors, in expression of CD95 on blood monocytes from CHC patients treated with INFa show an important role of cytokines system activation and mechanisms of programmed cell death in pathogenesis of chronic HCV infection. PMID- 11899824 TI - [Assessment of survival and risk of hepatocellular carcinoma in patients with cirrhosis of mixed (viral, alcoholic) etiology]. AB - AIM: To determine factors affecting overall survival and risk to develop hepatocellular carcinoma (HCC) in patients with hepatic cirrhosis (HC) of mixed (viral, alcoholic) etiology. MATERIAL AND METHODS: Mono- and multi-variance analysis of prognostic effects of such factors as age of the patients, mixed infection VHB/VHC, markers of HBV replication, antibodies to nuclear antigen of HBV (HBcAb) without HBs-system in the serum ("isolated" HBcAb), duration of viral infection, alcohol intake and abuse, dilatation of the esophageal veins, some laboratory parameters were studied in 55 HC patients having at least one marker of hepatitis B virus (HBV) and/or hepatitis C virus (HCV), long history of alcohol abuse. RESULTS: It was found that risk of HCC was associated with duration of alcohol abuse and infection, mixed HBV/HCV infection, age 60 and older. Of independent significance was only duration of alcohol abuse. Lethal outcomes in HC patients in the mixed infection were due to development of HCC (36%) and HC complications (64%). Survival of the patients was less in severe dilatation of the esophageal veins, high clinicolaboratory index, low level of serum albumin, presence of "isolated" BcAb and mixed viral infection. CONCLUSION: Of the greatest prognostic efficacy in respect of survival was the model combining "isolated" HBcAb, the degree of esophageal veins dilatation and hepatitis activity. PMID- 11899825 TI - [Clinical and demographic characteristics of various types of arterial hypertension in elderly patients (based on results of Russian scientific practical program ARGUS]. AB - AIM: It is the fragment of the trial aimed at the study of demographic indices and components of cardiovascular risk in patients over 55 with known and newly diagnosed arterial hypertension (AH) by referral to outpatient clinic depending on AH type. MATERIAL AND METHODS: For a week, 140 therapists from 14 regions of the Russian Federation measured AP in all persons aged 55 and older visiting outpatient clinics. A total of 5582 persons were examined. 3847 of them were already diagnosed to have hypertension. Primary diagnosis of hypertension was made in 5.7 examinees out of 2442. RESULTS: Patients with previous diagnosis of hypertension vs those with primary diagnosis had significantly greater index of body mass, had more often overweight, obesity and diabetes mellitus. Isolated systolic AH was encountered in 56% of all the primary cases. High and very high risk to develop cardiovascular complications was encountered in 59.9% of primary patients and 85.8% of those previously diagnosed. CONCLUSION: Patients aged 55 and older with newly diagnosed isolated systolic AH had the following most common factors of risk of cardiovascular complications: overweight, obesity, smoking, dyslipidemia, diabetes mellitus. In treated patients these risk factors are: overweight, obesity, dyslipidemia, diabetes mellitus, smoking. PMID- 11899826 TI - [Current status and perspectives in gastroenterology]. PMID- 11899827 TI - [Significance of pharmacological blockade of angiotensin II type I receptors for correction of abnormal 24-hour blood pressure profile depending on its variability in patients with arterial hypertension stage II]. AB - AIM: To specify variable 24-h arterial hypertension (AH) stage II profile and to assess significance of pharmacological block of the end of the renin-angiotensin aldosteron system for correction of the determined disorders. MATERIAL AND METHODS: The study was made of 46 men (mean age 42.8 +/- 3.28 years) with stage II AH and 25 normotensive controls (mean age 39.2 +/- 3.10 years). Depending on the magnitude of mean 24-h AP variability (APV), hypertensive patients were divided into two groups. Variability of systolic and/or diastolic AP (SAPV and DAPV, respectively) was considered high in at least 15.2 and/or 12.3 mm Hg variability, respectively, and normal at less values. RESULTS: AP 24-h profile in men with AH stage II and high APV compared to patients with normal APV is characterized by higher frequency of AP rise and less frequency of its night fall. In patients with high APV the drug eprosartan (teveten) is more effective in correction of hypertension and night fall of AP. CONCLUSION: Eprosartan has an adequate corrective activity in relation to absolute values of SAP and DAP in different hours. The highest hypotensive activity of the drug was seen in persons with initially high circadian AP variability within 24 hours. PMID- 11899828 TI - [Localization of vessel lesions in arteriosclerosis and blood lipid composition]. AB - AIM: To investigate correlations between composition of plasmic lipid fractions in patients with ischemic heart disease (IHD) and those with cerebrovascular insufficiency (CVI) caused by atherosclerosis. MATERIAL AND METHODS: 75 patients were divided into three groups; 26 patients with IHD free of CVI (group 1), 22 patients with CVI free of IHD (group 2), 27 patients with IHD and CVI (group 3). Blood lipids were measured by a standard mesiautomatic method using Technicon-AA 2 unit (USA). RESULTS: Hyperlipidemia (HLE) type II was most frequent in group 1 while HLE type IV or hypoalphalipoproteinemia without rise in cholesterol or triglycerides--in groups 2 and 3. CONCLUSION: In IHD without CVI dyslipidemia in most cases was associated with one of the additional risk factors (hypertension, smoking, diabetes mellitus) while in CVI it combined with two or three additional risk factors. PMID- 11899829 TI - [Immediate and long-term results of implantation of the coronary stent "BioDiamond"]. AB - AIM: To assess immediate and long-term outcomes after BioDiamond-stent implantation in patients with native coronary artery disease. MATERIALS AND METHODS: The BioDiamond stent was implanted in 112 patients with 132 de novo lesions. Most patients (54%) had unstable angina, 33% of the lesions were of type B2-C. RESULTS: No stent deployment failure occurred as well as acute or subacute stent thrombosis. The 6-month angiographic follow-up was obtained in 108 patients (98%) (125 lesions). The loss index was 0.40 +/- 0.27. Angiographic restenosis rate occurred in 14%. Repeat target lesion revascularization was successful in 16 patients with restenosis. CONCLUSION: The results of this study indicate a potential benefit of BioDiamond for prevention of stent thrombosis and restenosis in these relatively high-risk patients. PMID- 11899830 TI - [Viable myocardium: comparative evaluation of surgical and pharmacological treatment of patients with ischemic heart disease, post-infarct cardiosclerosis and chronic cardiac failure]. AB - AIM: To compare surgical and drug therapies of viable myocardium in IHD patients with left ventricular dysfunction (LVD), to show how to choose between them basing on the presence or absence of viable myocardium in the dyssenergy zones in such patients. MATERIAL AND METHODS: The study enrolled 55 IHD patients with postinfarction cardiosclerosis, left ventricular ejection fraction under 35%, circulatory disorder class I-III (NYHA classification). Coronary artery bypass operation (CABO) was made in 24 patients, 31 patients received drugs. The examination included dobutamin echo-CG, perfusion myocardial scintigraphy. Viability of the myocardium was assessed before and 6 months after the treatment. RESULTS: The patients were divided by the treatment and number of segments of viable myocardium: group 1--patients with viable myocardium subjected to CABO, group 2--patients with non-viable myocardium subjected to CABO, group 3--patients with viable myocardium treated with drugs, group 4--patients with non-viable myocardium treated with drugs. The greatest rise in LV EF, reduction of the end diastolic volume, end systolic volume of the left ventricle and functional class was observed in group 1. In group 2 these parameters changed less than in group 3. Patients of groups 2 and 4 were similar by parameters of LV global contractility. CONCLUSION: Improvement of central hemodynamics and functional class was the greatest in IHD patients after CABO having viable myocardium. If myocardium is not viable surgical treatment has no advantage over conservative in improvement of the pump cardiac function and is inferior to pharmacological treatment in effect on myocardium function in patients with viable myocardium in dyssynergia zones. PMID- 11899831 TI - [HindIII DNA-polymorphism of lipoprotein lipase gene in elderly patients with ischemic heart disease]. AB - AIM: To determine incidence of HindIII alleles of lipoprotein lipase (LPL) in Russian elderly patients with stable effort angina (SEA) functional class II-III regarding lipid metabolism. MATERIAL AND METHODS: Genotyping by LPL gene was performed in 103 patients with SEA. Of them 13 patients survived myocardial infarction (MI), 29 patients had diabetes mellitus. RESULTS: Incidence of alleles of HindIII DNA-polymorphism of LPL gene both in healthy and IHD patients is comparable with that in the West European populations. CONCLUSION: Genotype H+H+ of LPL gene is one of the markers of predisposition to MI, while allele H- is one of the resistance marks. PMID- 11899832 TI - [Combination of heterozygote blood coagulation factor V Leiden carriage with erythrocytosis as a cause of the hip deep vein thrombosis]. AB - AIM: To reject one of the variants of inherited thrombophylia in a 64-year-old patient with deep thrombosis of leg veins and high hemoglobin and red cell levels. MATERIAL AND METHODS: The study was made of antithrombin III and protein C, protein S levels; resistance to activated protein C; molecular structure of DNA coding factor 5; methylenetetrahydrofolate reductase. RESULTS: The patient was diagnosed to have heterozygote factor V Leiden mutation. The replacement of arginine by glutamine in position 506 of factor V molecule raises the risk of thrombosis. This risk was aggravated by high hemoglobin, red cells, hematocrit, low volume of circulating plasma, smoking. The patient had normal levels of leukocytes and platelets, normal spleen size, slightly lowered level of erythropoietin. CONCLUSION: The presence of thrombosis in patients with erythremia or erythrocytosis rejects one of the thrombophilia forms. PMID- 11899833 TI - [Idiopathic cardial achalasia (etiology, pathogenesis, endoscopic differential diagnosis, and treatment)]. PMID- 11899834 TI - [Current methods of treatment of diarrhea]. PMID- 11899835 TI - [Cardiovascular complications in mineralocorticism syndrome]. PMID- 11899836 TI - [Gold preparations in the treatment of rheumatoid arthritis]. PMID- 11899837 TI - [Dyspepsia and gastroesophageal reflux in adolescents]. AB - AIM: To study the prevalence and risk factors for dyspepsia and gastroesophageal reflux (GER) among adolescents. MATERIAL AND METHODS: Senior schoolchildren (forms 9-11) of four randomly selected secondary schools of Novosibirsk participated in the study (189 boys and 260 girls aged 14-17 years). They filled in the Bowel Disease Questionnaire, and sera were tested for antibodies against Helicobacter pylori infection. RESULTS: Dyspepsia was reported by 21% of the schoolchildren, being more frequent in girls (25%) than boys (14%, p = 0.004). GER occurred in 22% of adolescents with the same frequency in both genders. The combination of dyspepsia with GER was found in 8%. The prevalence of H. pylori infection was 56%. No association was found between the infection and abdominal symptoms, except heartburn. Among factors associated with dyspepsia were female gender, irregular meals, family history of dyspepsia. GER was related to H. pylori infection, obesity, smoking and irregular meals, 46% of adolescents with dyspepsia and 36% of those with GER have consulted a physician, and school absenteeism was reported by 55 and 40%, respectively. CONCLUSION: Dyspepsia and gastroesophageal reflux are widespread among the adolescent population and result in frequent use of health care resources. PMID- 11899838 TI - Space-age medicine, stone-age government: how Medicare reimbursement of telemedicine services is depriving the elderly of quality medical treatment. AB - We have the technology. What is needed is government financial commitment, so argues Kristen Jakobsen in the following discussion of "telemedicine." The term refers to the delivery of health care services by means of modern telecommunications technology. According to Ms. Jakobsen, the telephone, the fax machine, the Internet, and interactive audio-visual transmissions hold the key to making medical care more accessible and less expensive. Potential beneficiaries include vast populations of elderly in rural areas, who tend to be remote from upscale health care facilities and in need of the wherewithal to reach them. Standing in the way, in Ms. Jakobsen's view, is a government which lacks the boldness and the vision to lay an adequate fiscal foundation for this promising possibility. PMID- 11899839 TI - [Fluorescence diagnostics of premalignant and malignant changes in patients with Barrett's esophagus: doubts and hopes]. AB - Increasing incidence of esophageal adenocarcinoma that mostly arises from Barrett's esophagus (BE) has been observed. Therefore sensitive methods for early detection of dysplasias and adenocarcinomas in BE are more and more necessary. Fluorescence endoscopy is an alternative or complementary to standard four quadrant biopsies technique. The basics of diagnostics application of tissue autofluorescence and exogenous dyes fluorescence such as protoporphyrin IX, induced by 5-aminolevulinic acid are summarized. There are two methods of fluorescence diagnostics of dysplasia in BE: point fluorescence spectroscopy and real-time fluorescence imaging. Fluorescence spectroscopy is sensitive and specific but can be targeted only toward lesions visible in white-light endoscopy. Fluorescence imaging allows to examine large areas for occult foci of dysplasia but selectivity is somewhat limited by the background fluorescence of Barrett's mucosa. Laser induced fluorescent endoscopy (LIFE) images of esophageal dysplasia are presented. Up-to date results of fluorescence spectroscopy and fluorescence imaging of BE are reviewed. These results indicate fluorescence diagnostics promising and clinically useful for detection of early adenocarcinoma and dysplasia in BE. PMID- 11899840 TI - [Tricyclic antidepressants as inhibitors of brain glutathione-S-transferase]. AB - Tricyclic antidepressants, amitriptyline, doxepin--derivatives of cycloheptadiene as well as imipramine and clomipramine, derivatives of dibenzazepine inhibit the activity of glutathione-S-transferase pi isolated from different regions of human brain (parietal cortex, frontal cortex, brain stem). The inhibitory effect of studied drugs depends more on their chemical structure than on brain localization of the enzyme. All tricyclics bind nonspecifically to the effector site of glutathione-S-transferase (GST). The obtained results indicate that the inhibitory effect of tricyclic antidepressants on brain GST may decrease the efficiency of the enzymatic barrier, which protects brain against toxic electrophiles, and contribute in their adverse effects. On the other hand, brain GST may decrease the therapeutic effects of tricyclic antidepressants by binding them as ligands. PMID- 11899841 TI - [Usefulness of preoperative assay CEA and CA 19-9 in colorectal cancer patients]. AB - Carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) are well known tumor markers expressed by colorectal cancer (CRC), particularly in advanced cases. In this study serological expression of CEA and CA19-9 concerning different clinicopathological factors and sensitivity of diagnosis of CRC by combination of both markers were evaluated. Eighty one patients with diagnosed CRC were in this study included. According to Dukes' classification there were 11 in stage A, 34 in stage B, 17 in stage C and 19 in stage D. Blood samples were collected before operation. CEA and CA19-9 were determined by MEIA (normal range 0-3 ng/ml for CEA and 0-37 U/ml for CA19-9). The statistical analysis revealed that the CEA well correlated with histological type, liver metastasis and term of symptoms. The CA19-9 well correlated with liver metastasis, peritoneal dissemination, nodal involvement and cancer localization. The levels of CEA and CA19-9 increased with stage of the tumor, but only in stage D the difference was statistically significant (for CEA p = 0.005, for CA19-9 p = 0.039). At time of diagnosis 50.6% of the patients had elevated serum levels of CEA and 29.6% of CA19-9. In combination of both antigens this elevation was in 54.3% of CRC patients. The common use of CEA and CA19-9 was more efficacious in identification of patients at high risk. The combination assay of CEA and CA19-9 did not cause a significant increase of sensitivity in diagnosis CRC. PMID- 11899842 TI - [Monitoring of renal function in epileptic children and teenagers treated with valproic acid or carbamazepine in concomitant therapy with tiagabine]. AB - The aim of the study was the evaluation of the influence of 4-month concomitant tiagabine (TGB) and valproic acid (VPA) or carbamazepine (CBZ) therapy on renal function of epileptic children and teenagers. Initial parameter values, indicated on renal disfunction, were compared with these obtained after VPA and TGB or CBZ and TGB therapy and with values in healthy children and teenagers. Investigation group was composed of 22 children and teenagers with drug-resistant focal epilepsy. We observed that in the time of concomitant VPA and TGB therapy increased the NAG/g creatinine activity index. In spite the fact of statistical significance of these changes, they were not outside the normal range. beta 2 microglobulin concentrations in urine of epileptic children treated with VPA in monotherapy before concomitant therapy with TGB were higher than in control group. That difference was statistically significant. Addition of TGB to the therapy normalized this parameter. During concomitant VPA and TGB or CBZ and TGB therapy we didn't observe statistically significant changes of parameters indicating on glomerular disfunction. In the VPA therapy before concomitant treatment with tiagabine the disfunction of tubules and glomerules was observed. On the other side in the concomitant VPA and TGB therapy the disfunction of tubules and glomerules didn't occurred. We can conclude that concomitant therapy VPA or CBZ with tiagabine don't affect the renal function in clinical significant manner. Therapy with VPA could leads to minimal disfunction of tubules what is represented by increasing of beta 2-microglobulin level in urine. PMID- 11899843 TI - [Lipids peroxidation in platelets in patients with bladder cancer treated with Mycobacterium suspension]. AB - Platelet is blood's morphotic element in which intense energy metabolism takes place, which makes it possible to participate in the complex processes of the organism's homeostasis. The aim of the study was to analyse aerobic metabolism in the platelets, taking into consideration lipids peroxidation in patients with bladder cancer treated with the bacillus Calmette-Guerin (BCG) Mycobacterium suspension. The determination of superoxide dismutase (SOD-1) and malonyl dialdehyde (MDA) concentration activity constituted this evaluation's parameters. A group of 12 patients (4 women and 8 men) aged 54-67 years (average age 61) in which superficial bladder cancer was diagnosed were included in the study. Electroresection was carried out and subsequently, after 14 days, BCG Mycobacterium suspension was administered in intravesical instillations, in a 6 week cycle according to Morales. The material for the study was venous blood taken from the patients in three periods (before treatment, after the last clyster and 30 days after treatment) into the tubes with the addition of 1% EDTA in the ratio of 9 blood volumes to anticoagulant's one volume. Superoxide dismutase activity (Cu Zn--SOD) was determined according to Misra and Fridovich. The values were expressed in lamellar protein U/g protein. MDA concentration in platelet's TBARS was determined according to Pansa et al. MDA concentration included in TBARS was expressed in nmol/109 platelets. The controls were healthy volunteers in the same age range. In unaided studies a significant rise in superoxide dismutase (SOD-1) activity was obtained with the 1574.606 average before treatment > 2137.03 after treatment and 2646.4 after a month observation. Whereas MDA concentration increased in non-treated patients to 1.97, after treatment it dropped down significantly to 1.55 and sustained the downward trend after 30-day observation 1.4 nmol/109 platelets. The use of BCG intravesical clysters causes lipids peroxidation inhibition (decrease in MDA concentration) and the increase of SOD-1 activity results in smaller aggregation of platelets, preventing the formation of neoplastic metastases. PMID- 11899844 TI - [Autoimmune thrombocytopenia in chronic liver disease]. AB - Thrombocytopenia (TP) often accompanies chronic liver diseases. The causes are numerous and include impaired production of blood platelets, spleen sequestration, and the immune factors, e.g. antiplatelet autoantibodies. ELISA (GTI-PAKPLUS) examinations were conducted in order to estimate the rate of autoimmune thrombocytopenia occurrence in patients with thrombocytopenia in the course of chronic hepatitis (10 patients) and liver cirrhosis (20 patients). Blood platelet activity was also evaluated as well as the expression of platelet glycoproteins (GPIIb, GPIIIa, and GPIX) in platelets of the patients and the controls. It was observed that autoimmune TP occurred in 30% of patients with liver cirrhosis and in 10% of patients with chronic hepatitis, in which anti GPIIb/IIIa, GPIa/IIa, and HLA class I antibodies were detected. In all patients there occurred significant/marked platelet activation with CD61P expression. Thrombocytopenia in patients showed a similar activity after thrombin stimulation to that in healthy individuals. Expression on GPIIb platelet receptors was markedly increased and GPIX decreased in patients in comparison to the controls. There was no correlation between the occurrence of certain types of anti-platelet autoantibodies and the expression of GP on thrombocytes in these patients. PMID- 11899845 TI - [The relation between Chlamydia pneumoniae infection and abdominal aortic aneurysm]. AB - The aim of our study was to evaluate the frequency of C. pneumoniae infection in abdominal aortic aneurysm (AAA) patients by measuring C. pneumoniae specific serum IgG, IgM and IgA levels and the activation of their immune system by measuring the concentrations of IL-10, IL-12, IFN-gamma and TNF-alpha in patients' serum. Microimmunofluorescence method was applied to evaluate the level of anti-C. pneumoniae IgG, IgA and IgM. The concentrations of cytokines were evaluated using ELISA method. Serologic markers of persistent C. pneumoniae infection have been detected in 25/28 (89.3%) patients and in 6/20 (30%) healthy controls. In 40% (10/25) of patients with serologic markers of persistent C. pneumoniae infection high titers of specific IgG and IgA indicated active infection--reinfection or exacerbation of chronic infection. Mean concentrations of IL-10, IL-12, IFN-gamma and TNF-alpha indicated lack of protection against intracellular pathogens. Since all patients in this group were diagnosed as having symptomatic AAA, we suggest that active infection can exacerbate inflammation in the AAA wall and accelerate progression of the disease. In our opinion patients with active C. pneumoniae infection may be candidates to the antimicrobial treatment. PMID- 11899846 TI - [Evaluation of glucose, insulin, C-peptide uric acid serum levels in patients with psoriasis]. AB - The aim of our study was to estimate the serum levels of glucose, insulin, C peptide and uric acid in patients with psoriasis before and after treatment. The study included 12 males with active form of psoriasis and 15 control subjects carefully matched to the psoriatic patients for age and BMI. All measured parameters were in patients with psoriasis significantly increased and dependent on the BMI. Compared with pretreatment values of glucose and uric acid were significantly lower during therapy. The increase in the mean C-peptide and insulin levels after psoriasis therapy was constant and independent from clinical stage of disease. The results of the present study have provided evidence for the importance of impaired glucose and purine metabolism in patients with psoriasis in the increase risk of development of diabetes mellitus and hypertension. PMID- 11899847 TI - [Multimodal evoked potentials in malabsorption syndrome]. AB - The multimodal evoked potentials (visual, somatosensory and auditory brainstem) in 23 patients with malabsorption syndrome of different origin were investigated. The diagnosis of the disease was confirmed on the basis of histological examination and result of D-ksyloza test. The control group consisted of 30 healthy persons. Examination of visual evoked potentials revealed significant prolongation of latency of P 100 component in examined group in comparison with controls. Latency of N13 and N20 of somatosensory evoked potentials in patients with malabsorption syndrome were also significantly prolonged when compared to controls, otherwise transit time to cortex (TTC) was merely prolonged. Auditory brainstem potentials were also abnormal. Changes involved prolongation of latency of I, III and V responses and prolongation of interpeak latencies of I-III, III V, I-V as well. The amplitudes of the examined evoked potentials between patients and controls did not differ significantly. On the basis of obtained results in was pointed out that different specific afferent systems are affected in patients with malabsorption syndrome what seems to be connected with vitamin's deficiency, especially B12 and E. Authors conclude that multimodal evoked potentials examinations are useful in the diagnosis and monitoring of the disease, specially in subclinical cases. PMID- 11899848 TI - [Focal nodular hyperplasia of the liver in a young female]. AB - Focal nodular hyperplasia (FNH) of the liver is a lesion characterized by a well circumscribed region of hyperplastic liver tissue with stellate fibrosis. The pathogenesis of FNH is unknown but various authors consider that this lesion may be a response to a preexisting vascular abnormality. A 27-year-old woman was referred because of large liver lesion detected by ultrasound abdominal examination. Doppler ultrasound, computed tomography and magnetic resonance suggested this was FNH. The patient was conducted to resection of the tumor because of size of the tumor and presence of clinical signs and symptoms. Pathological examination of the surgical resection confirmed diagnosis of FNH. Follow-up after 1 and 2 years showed that the patient remained well but she complained of general weakness and we found unexplained elevation of GGT. Liver biopsy was performed 1 year after resection of the tumor and histopathological examination showed only minimal reactive changes. PMID- 11899849 TI - [Hypersensitivity of estrogen receptors as a cause of gigantomasty in two girls]. AB - The authors present two girls with gigantomastia, 14 and 15 years of age. Laboratory examinations demonstrate an increase of estrogen receptors in the glandular tissue. In the immunohistochemical investigations ascertained was a receptor hypersensitivity of estrogen and progesterone receptors. In one of the girls shown was hyperprolactinemia in the metoclopramide test (patient J.K.) In the physical examination hyperlordosis, kyphosis, gigantomastia was observed. Additional in patient K.S.--anorexia and patient J.K.--Sjogren's syndrome. PMID- 11899850 TI - [Cytokines in rheumatoid arthritis. I. Proinflammatory cytokines]. AB - Proinflammatory cytokines play an important role in the pathogenesis of rheumatoid arthritis. It is why they became the targets for new therapies. In this review we describe their expression in synovial tissue, synovial fluid and in serum, and correlation with disease activity. Particular attention was paid to the possibilities of the alternative treatment strategies modifying the balance of cytokine network, in the rheumatoid arthritis patients, towards limiting their proinflammatory activity. Inhibiting the action of proinflammatory cytokines by using their specific inhibitors or anti-inflammatory cytokines have shown significant clinical benefits with mild side effects. PMID- 11899851 TI - [Cytokines in rheumatoid arthritis. II. Anti-inflammatory cytokines]. AB - In this review the role of anti-inflammatory cytokines in the pathogenesis of rheumatoid arthritis is presented. We describe their expression in synovial tissue, synovial fluid and in serum and correlation with disease activity. Special attention was paid to the possibilities of the alternative therapies modifying the balance of cytokine network, in the rheumatoid arthritis patients, towards anti-inflammatory state. Several such approaches have shown significant clinical benefits with mild side effects. PMID- 11899852 TI - [The role of lipids and lipoproteins in acute pancreatitis]. AB - The aim of this work is to evaluate acute pancreatitis (AP) including specially role of lipid's and lipoprotein's metabolism disorders. On the ground of actually literature we take up studies to answer for the question: lipid's and lipoprotein's metabolism disorders are the cause or the effect of AP, or whether they are a phenomenon taking place independently on the disease? Our analysis indicate an immediate relation between lipid's metabolism and acute pancreatitis, but don't permit to table univocals motions. To this it's necessary removal examinations on the large and various group of patients with acute pancreatitis. PMID- 11899853 TI - [The methods of estimation and monitoring liver fibrosis in patients with chronic viral hepatitis]. AB - Liver fibrosis may be defined as increased deposition of extracellular matrix components which is a result of the dynamic imbalance between synthesis and degradation of connective tissue. This pathological process is caused by chronic liver injury e.g. chronic infection of hepatotropic viruses, alcohol intake and autoaggressive liver diseases. Author presents methods' review of estimation liver fibrosis in patients with chronic hepatitis with special reference to the role of serum liver fibrosis markers. PMID- 11899854 TI - [Restless legs syndrome]. AB - The authors present recent data about epidemiology, pathogenesis and clinical features of idiopathic and symptomatic restless legs syndrome. A new aspects of pharmacological treatment and modification of daily activity are also described. PMID- 11899855 TI - [Endolymphatic therapy]. AB - Using physiological function of lymphatic vessels and nodes and also a fact that they are a frequent place of spreading of diseases, there are attempts of endolymphatic therapy--it means drug infusions into lymphatic vessels for the purpose of penetrate lymphatic nodes engaged by disease. There are more and more attempts of using the cytostatics placed in liposomes in endolymphatic. PMID- 11899856 TI - [Changes in the endocrine system in the course of sulpiride therapy]. AB - The influence of sulpiride, a neuroleptic included into the group of benzamide derivatives, upon endocrine system has been discussed in the article. It is noted that sulpiride clearly increases the evolution of prolactin in both sexes. It gives a chain of various consequences. They are, among others, impairment of gonad activity in both sexes which causes emmeniopathy and galactorrhea in women and impotence in men. The influence of hyperprolactinemia, caused by sulpiride, upon the level of testosterone in plasma and also upon the evolution of progesterone has been also discussed. The knowledge of the influence of sulpiride upon endocrine system is necessary for physicians applying therapy by this medicine. In the article the analysis has been carried out how wider and wider used neuroleptic-sulpiride--influences upon endocrine system and particularly upon the evaluation of prolactin. In the article the main methods of action of the known neuroleptic-sulpiride have been discussed. This is an antiautistic, activizing, antipsychotic and antidepressive action. The other authors used sulpiride in therapy of depressive endogenic syndrome. They obtained a good therapeutic effect. The drug distribution in human organism was also taken into consideration. The mechanism of action of sulpiride and its extrapsychiatric application was also mentioned. The other authors often combines the drug with benzodiazepine derivatives which soothe the fear characteristic of depression. PMID- 11899857 TI - [Treatment strategies of withdrawal from long-term use of anabolic-androgenic steroids]. AB - Anabolic-androgenic steroid (AAS) withdrawal is established to be an important, though poorly known medical problem, because of AAS potency to cause physical and psychological dependence. Thus discontinuation of high-dose, long-term anabolic steroid use, apart from endocrine dysfunction (hypogonadotropic hypogonadism), may lead to development of withdrawal symptoms. They include mood disorders (with suicidal depression as the most life-threatening complication), insomnia, anorexia, decreased libido, fatigue, headache, muscle and joint pain, and desire to take more steroids. The withdrawal from anabolic steroids usually requires treatment. Clinical management, as with other drugs of abuse, consists of supportive therapy and pharmacotherapy. The goals of treatment are to restore endocrine (hypothalamic-pituitary-gonadal, HPG) function and to alleviate withdrawal symptoms. The endocrine medications that are targeted specifically to ameliorate HPG function include testosterone esters, human chorionic gonadotropin, synthetic analogues of gonadotropin-releasing hormone and antiestrogens. They are indicated in the presence of persistent clinical symptoms or/and laboratory evidence of HPG dysfunction. Other medications, that are targeted to provide symptomatic relief include antidepressants (especially serotonin selective re-uptake inhibitors), nonsteroidal anti-inflammatory drugs and clonidine. Notwithstanding, it should be remembered that many of the above mentioned drugs have their own potential for abuse or side-effects, so their use must be carefully weighted and optimal treatment strategies for AAS withdrawal must await further clinical research. PMID- 11899858 TI - [The role of vitamin C in absorption and dismissing of ions of metals]. AB - There is growing interest in the physiological role of vitamin C. In this paper L ascorbic acid is discussed in detail, including its chemical properties and biological activity. The primary goal of this review is to present the chemical characteristic of this compound and to discuss its relation to various biological functions of vitamin C in absorption and to dismissing of ions of metals. PMID- 11899859 TI - [Objects and evolution courses of modern teratology. Report from the 41st Annual Meeting of the Teratology Society--June 23-28.06.2001--Montreal. Canada]. AB - This report refers to the actual opinions on epidemiology, pathogenesis and prevention of congenital malformations presented during the 41st Annual Meeting of Teratology Society, held on the 23rd-28th June 2001 at Montreal in Canada. PMID- 11899861 TI - [Assessing the relationship between the air pollution and incidence of respiratory diseases in the Primor'e territory]. AB - The prevalence of respiratory diseases in the Primorye territory is discussed. Ecological risk of air pollution effects on respiratory morbidity is estimated. High ecological risk of respiratory diseases in the cities of the region is determined by car transport waste discharge. Children and adolescents are more sensitive to air pollution and more often suffer from respiratory diseases. PMID- 11899860 TI - [Relationship between emotional stress in female residents of the city of Chapaevsk and toxicological and genetic values]. AB - The severity of stress was evaluated in 45 female residents of the town of Chapaevsk (3 groups, 14-15 females each, with different levels of exposure to dioxins) using 5 standard psychological questionnaires. The results of testing were correlated to plasma dioxin concentrations and levels of chromosome aberrations in peripheral blood lymphocytes of the same donors. The results indicate that the groups differed by the degree of stress, the highest level being observed in women occupationally exposed to dioxins. The severity of stress correlated with plasma dioxin concentrations (p < or = 0.001) and levels and spectra of chromosomal aberrations in peripheral blood cells (p < or = 0.05, p < or = 0.01 in different tests). Presumably, one of the causes of high level of chromosomal aberrations in these women is dioxin-induced decrease of the adaptation potential, resulting in increase of the genome sensitivity to genotoxic factors of other origin). PMID- 11899862 TI - [Ecological and hygienic evaluation of microbiological process in the soil contaminated by anion surfactants and heavy metals]. AB - Disorders in microbiocenosis and biological activity of different types of soil were studied in a laboratory model experiment and the survival of reference and pathogenic microorganisms exposed to sulfonol and lead was evaluated. The most sensitive tests for such studies are evaluation of the number of nitrifiable bacteria, nitrifying, proteolytical, dehydrogenase, and cellulase activities, and survival of Salmonella and lactose-positive Escherichia coli in soil. PMID- 11899863 TI - [Incidence of pyelonephritis on territories with different levels of anthropogenic loading]. AB - Relationship between atmospheric pollution with harmful substances and incidence of pyelonephritis was studied for the populations living in ecologically unfavorable and clean regions of an industrial city. Disease intensity and incidence were significantly higher in polluted regions, the probability of error being no more than p < 0.05-p < 0.001. The correlation between atmospheric nitrogen dioxide concentration and pyelonephritis incidence was the strongest for all years analyzed. Correlation coefficients tend to increase: during the analyzed period they increased from Rxy = 0.42 +/- 0.4 (p < 0.01) to Rxy = 0.64 +/- 0.15 (p < 0.01). PMID- 11899864 TI - [Contribution of heavy metals to the development of reproductive disorders]. AB - In order to evaluate the role of heavy metals in the reproductive status, a complex hygienic evaluation of total daily entry of lead, cadmium, zinc, and copper in the organism, biomonitoring of metals in indicator biosubstrates, and epidemiological follow-up of the reproductive function were carried out in 6962 healthy women living in industrial cities of the Dnepropetrovsk region. Total daily entry of lead is 0.27 mg, that of cadmium 0.029 mg, which meets the WHO requirements. However the entry of copper and particularly zinc is 1.5-3 times below the physiological norm. Biomonitoring showed high concentrations of lead and cadmium in the body. Regular entry of abiotic metals even in low concentrations in the presence of deficiency of essential metals is fraught with risk of reproductive diseases, which was proven mathematically for all stages of the reproductive function: gestation, labor, and neonatal period. PMID- 11899865 TI - [The leading occupational medicine journal "Gigiena i Sanitariia" (Hygiene and Occupational Medicine) is 80 years old]. PMID- 11899866 TI - [Infectious diseases in children living under conditions of technology-related pollution of atmospheric air]. AB - Epidemiological and clinical studies in children with acute respiratory viral infections (ARVI) and viral hepatitis A (HVA), living in zones with different levels of technogenic pollution of the atmosphere, showed that the incidence and clinical course of viral infections in children depended on the technogenic pollution of the environment, this effect being the more pronounced, the higher the level of xenobiotics in the air. Children living under conditions of high technogenic pollution of the environment should be regarded as a group at risk of more severe ARVI and HVA with complications. PMID- 11899867 TI - [Effects of ultraviolet radiation on Cryptosporidium oocysts and Lamblia cysts in drinking water]. PMID- 11899868 TI - [Hygienic evaluation of actual nutrition and health status of sales personnel in the city of Artem]. AB - Actual nutrition of saleswomen was studied in the town of Artem (Primorye territory), characterized by unfavorable demographic values. Fifty-two women aged 30-39 years were examined, all of them living in the region for at least 3 years and engaged in foodstuff sales for more than 3 years. Their nutrition was studied by 24-hour reproduction of nutrition. In addition, nutrition regimen, food preferences were evaluated and alimentary-dependent diseases were detected. The study revealed an imbalance in the nutrition structure, with low consumption of meat and fish, fruits and vegetables, vegetable oil and high consumption of sugar, potato, and bread. The consumption of the main nutrients and micronutrients was insufficient, energy value of food was low, and meals hours irregular. PMID- 11899869 TI - [Rationale-based nutrition as a factor of anemia prevention in pregnant women]. AB - Rational nutrition, based on properly arranged regimen including quantitative and qualitative distribution of the daily ration into meals is one of important elements in correction of anemia, endemic for the West Siberian region. Pregnant women should be referred to a group at risk of this disease; special nutrition standard, therapeutic and prophylactic rations should be created for them with due consideration for communal conditions, health status, and their actual nutrition. PMID- 11899870 TI - [The role of bioactive food additives in therapeutic and prophylactic nutrition]. AB - The efficiency of therapeutic and prophylactic nutrition (TPN) is determined by its scientific basis. The diets should be created with consideration for pathogenetic mechanisms of the effects of occupational hazards, energy expenditures, ecology of the habitat, national features of nutrition, and unfavorable factors associated with the risk of the underlying and concomitant diseases. Use of bioactive food additives (BFA) essentially facilitated the development and utilization of TPN under conditions of hazardous production and unfavorable ecology. The technology of making a perspective BFA Vitagmal has been developed. Biomedical evaluation of this BFA was carried out: protein, carbohydrate, and mineral metabolism, morphology of organs and tissues in experimental animals fed Vitagmal, and its effects on clinical and physiological status of man were studied. The effect of BFA on the metabolism was studied in patients with metabolic alimentary obesity. The results demonstrated a pronounced health-fortifying antioxidant effect of Vitagmal, which had a favorable impact on cardiovascular and central nervous functions, stimulated the metabolic processes, had a slight "protective effect" in ethanol intoxication, and no side effects. This recommends Vitagmal for wide use in therapeutic and prophylactic medicine. PMID- 11899871 TI - [Hygienic evaluation of the efficiency of milk fluorination for the prevention of dental diseases]. PMID- 11899872 TI - [The formation of the basis for the health life style of school children]. AB - Sanitary knowledge of graduates of the Tashkent schools on healthy life style was studied. Poor knowledge and understanding of the value of health and role of healthy life style in health preservation were revealed. The majority of graduates just vaguely know the main components of a healthy life style, as a result of which their actual life style cannot help them preserve their health. The authors suggest that a curriculum for the formation of a healthy life style be developed for schoolchildren, which will determine the priorities and the most rational forms of a healthy life style. PMID- 11899873 TI - [Medico-social aspects in the prevention of viral hepatitis B and C in young people and adolescents]. AB - The prevalence of viral hepatites B and C (HVB and HVC) among young people and adolescents in the Astrakhan region was evaluated. During recent 5 years, the incidence of HVB increased from 27.3 to 44.14 cases (1994-1999) per 100,000 population, that of HVC from 1.6 to 24.4 cases per 100,000. The majority of patients were young people and adolescents at the age of 15-29 years (67.1% HVB and 70.2% HVC). Increased disease incidence in this group is explained by activation of sexual transmission and increase in the number of patients infected through intravenous drug use. Problems in prevention of HVB and HVC are discussed. PMID- 11899874 TI - [Adaptation of junior medical students to the studying process]. AB - The characteristics of the main nervous processes, intellectual working capacity, autonomic regulation of cardiac rhythm and their relationship with progress in studies were evaluated in students. Good students were characterized by higher mobility of nervous processes, lower strain of cardiac rhythm regulation, and more economic cardiovascular work. PMID- 11899875 TI - [Toxico-genetic risk factors for adolescents living in a large industrial city]. AB - Cytogenetic study of adolescents in a large industrial city (n = 211) and donors (n = 41) showed an increase in the number of chromosomal aberrations per 100 lymphocytes in urban adolescents from 1.58 +/- 0.41 in 1992 to 4.53 +/- 0.48 in 1996. A significant increase in the spontaneous level of aberrations (2.02 +/- 0.21) was observed starting from 1993. Time course of the number of chromosomal aberrations and components of the environment (radiation background, chemical pollution of potable water, foodstuffs, and atmospheric air) in the city of Kemerovo in 1992-1996 are compared. According to the data of analysis of correlations, the most significant genotoxicants are nitrogen oxide (II), methanol, and benz(a)pyrene. PMID- 11899876 TI - [Toxicological and hygienic evaluation of sardor, a new defoliant agent and its occupational standardization]. AB - Study of the toxicity of Sardor, a new cotton defoliant, showed that it is a low toxic compound. Hygienic norms and regulations for its use in agriculture were developed. PMID- 11899877 TI - [Lead contamination of alcoholic drinks in crystal bottles]. PMID- 11899879 TI - [Efficiency of Diprim for the restoration of the functional status of the liver after its damage by ethanol poisoning]. AB - The effects of Diprim, a natural polyphenol complex isolated from Amur grapes crest, on lipid metabolism, lipid peroxidation (LPO), and antioxidative system of the liver were studied in ethanol-treated rats. Diprim injected to animals during ethanol abstinence arrested stress and thus inhibited lipolysis in fatty tissue and prevented triglyceride accumulation in the liver. Ethanol-suppressed metabolic reactions of the liver concerned with the synthesis of phospholipid fractions (phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine) were restored. Diprim promoted inhibition of LPO processes (the level of malonic dialdehyde decreased) and a pronounced glutathione-saving effect, which is largely due to its components--polyphenols which capture free radicals. PMID- 11899878 TI - [Effects of toxic substances on opportunistic microorganisms]. AB - Anti-immunoglobulin activity of immunoglobulins, rarely receiving attention in literature, occupies a special place among the special bacterial substances released by bacteria exocellularly. We studied the effects of some pollutants (phenol, diethanolamine, nitric and hydrochloric acids) on this pathogenicity factor. Phenol and hydrochloric acid added to tap water caused statistically significant changes in anti-immunoglobulin activity of Klebsiella strains. PMID- 11899880 TI - [Factors contributing to the pollution of the environment in a large industrial center]. AB - Many-year (1993-1998) comparative hygienic evaluation of industrial and car transport waste discharge in five regions of Samara, making use of systemic analysis methods, helped single out the most significant chemical compounds and develop a mathematical model of the effects of industrial waste in a large city on the development of bronchopulmonary diseases. PMID- 11899881 TI - [Ecological and hygienic evaluation of the lignin by-products utilization]. AB - Main trends in utilization of lignin wastes formed in complex wood processing are discussed. Toxicological and hygienic characteristics of lignins and products of their biotransformation and chlorination are presented. Ecology and hygiene of fertilizers, preserving agents, and construction materials based on lignins are evaluated. PMID- 11899882 TI - [On the methodology of social hygienic monitoring]. PMID- 11899883 TI - [Sanitary surveillance and sanitary statistics]. PMID- 11899884 TI - [Botulism cases in the Amur region]. PMID- 11899885 TI - [A method for the studying of the relationship between protein contamination of atmospheric air by enterprises engaged in microbiological synthesis and human health]. PMID- 11899886 TI - [The use of covariation analysis for evaluation of the relationship between atmospheric air pollution and external respiration function]. PMID- 11899887 TI - [Methodological approaches to hygienic assessment of iodine deficiency risk]. PMID- 11899888 TI - [Problems in monitoring air formaldehyde levels]. PMID- 11899889 TI - [Modern approaches to study and evaluation of viral contamination of drinking water]. PMID- 11899890 TI - [The evaluation of serum immunoglobulin level as a method for the evaluation of immunoreactivity in alcoholics, HBsAg carriers]. PMID- 11899891 TI - [Effects of dioxins in the development of malignant tumors and disorders of reproductive health of the population]. AB - The mortality caused by malignant tumors in the town of Chapaevsk, characterized by increased level of dioxins in the environment, is statistically higher than the expected values. For men the relative risk of general morbidity is 1.9 and mortality, 1.8; for lung cancer: morbidity, 3.3 and mortality 3.1; for urogenital cancer: morbidity, 3.6 and mortality, 2.6; for gastric cancer: morbidity 1.9 and mortality, 1.7. In women the morbidity and mortality due to breast cancer (relative risks 1.9 and 2.1, respectively) and cancer of the cervix uteri (relative risks 2.1 and 1.8, respectively) are increased. Changes in the reproductive health of residents of this town are as follows: high incidence of spontaneous abortions, appearance of small-for-date babies, and genital disorders in body (cryptorchidism, phimosis, hypospadia, delayed sexual development). PMID- 11899892 TI - Molecular incompatibilities in hemostasis between swine and men--impact on xenografting. AB - The rejection of xenografts is associated with vascular-based inflammation, thrombocytopenia and the consumption of coagulation factors that may evolve into disseminated intravascular coagulation. Natural regulators of coagulation in porcine xenografts have abnormal physiological interaction with human effectors. We have demonstrated the enhanced potential of porcine von Willebrand factor to associate with human platelet GPlb to be dependent upon the isolated AI domain of von Willebrand factor. The inability of porcine tissue factor pathway inhibitor (TFPI) to adequately neutralize human factor Xa (FXa), the aberrant activation of both human prothrombin and FXa by porcine EC and the failure of the porcine natural anticoagulant, thrombomodulin to bind human thrombin and hence activate human protein C may all be pathogenetic in the DIC following xenograft rejection. In this brief review, molecular incompatibilities of contributors in the physiologically fine tuned system of hemostasis are summarized and brought in context with the disordered thromboregulation, that seems to be invariably associated with delayed xenograft rejection. Possible therapeutic interventions are discussed. PMID- 11899893 TI - In vivo microscopic assessment of microcirculatory changes in a concordant xenogeneic primate experimental set up. AB - Organ transplantation is always connected with ischemia and thus reperfusion injury of the graft. One of the characteristics in this process is the temporary and permanent adherence of leukocytes to the endothelium of the graft. This cell to-cell interaction allows the immunocompetent cells to interact in the sense of antigen recognition with mainly defect endothelial cells. It was the aim to study whether induction therapy with poly-clonal ATG's would reduce or even prohibit these early interactions. MATERIAL AND METHODS: The distal extremities of cynomolgus monkeys were flushed via the femoral vessels and reperfused with ABO compatible human heparinised blood of a hct of 30%. Microcirculation was observed applying intra-vital microscopy. The images taken by a CCD-camera are recorded on video tapes for later off line evaluation. pATG 1 is directed against jurkat cells, pATG 2 against human thymocytes. In controls the blood vessels were perfused with untreated blood. In groups 2 and 3 the blood was treated with the amount of the ATG's used in clinical therapy 15 min. prior to perfusion. The total ischemia time was 1 hour. RESULTS: Five minutes after perfusion rolling was seen in the untreated animal, this increased to change into sticking after 30 min. The blood flow (RBC) in larger venoles remained almost normal. Both polyclonal ATG's inhibited the adhesion to a large extend. CONCLUSION: Ischemia reperfusion results in increasing adherence of leukocytes in the described model. pATG's suppress this phenomenon completely. This suggests that pATG's contain a number of antibodies directed against various types of cell surface molecules which are involved in reperfusion injury and that pATG's have a favourable influence on the early I/R-mechanisms after organ transplantation and a protective action when used as pre-operative induction therapy. PMID- 11899894 TI - Porcine endogenous retroviruses (PERVs): adaptation to human cells and attempts to infect small animals and non-human primates. PMID- 11899895 TI - Ex-vivo hemoperfusion (eHPS) of pig-lungs with whole human blood: effects of complement inhibition with a soluble C1-esterase-inhibitor. AB - OBJECTIVES: Xenotransplantation could be a future alternative to allotransplantation due to increasing organ shortage. Complement activation plays a major role in hyperacute rejection (HAR) in pig-to-human combinations. We developed an ex-vivo hemoperfusion (EHP) system to investigate pathophysiology of HAR in the lung. After standardizing the model, the effect of a soluble complement inhibitor (C1-INH, Berinert) was investigated. METHODS: Pig lungs were harvested following cold perfusion with Celsior preservation solution. EHP was performed using fresh heparinized human blood plus C1-INH (n = 6) or heparinized human blood as control (n = 4). Bloodgas analyses (BGA), pulmonalarterial pressure (PAP) were monitored. P-selectin and L-selectin were measured. Tissue samples were taken and microscopic changes evaluated. RESULTS: BGA, PAP, macroscopic and microscopic changes in the control group showed HAR, while the C1 INH group showed significantly longer function. Leucocytes and platelets were markedly activated in the control, whereas in the treated group L-selectin and P selectin values indicated lower activation. HE stainings showed maintained lung architecture after perfusion of pig-lungs with human blood plus C1-INH. Immunohistochemistry showed less C1q, C3, C5b-9 activation in the C1-INH group. CONCLUSIONS: Investigation of interaction of human blood with pig lung endothelium could be done in this model. C1-INH attenuates HAR in a pig-to-human lung transplantation model by decreasing the activation of adhesion molecules. C1 INH could play a role in induction therapy of future lung xenotransplantation. PMID- 11899896 TI - Review of a flat membrane bioreactor as a bioartificial liver. AB - Recent developments in tissue engineering permit to use isolated hepatocytes in a bioreactor for the creation of a bioartificial liver which supports patients suffering from acute liver failure. In this study, the authors discuss the development of a flat membrane bioreactor using pig hepatocytes for the replacement of liver functions. The flat membrane bioreactor permits a high density hepatocyte culture under sufficient oxygenation conditions, comparable to an in vivo microenvironment. In this bioreactor, built according to the in vivo organisation of the liver, pig hepatocytes are cultured with non-parenchymal cells within an extracellular matrix between oxygen-permeable flat-sheet membranes as individual plates. The performance of the "scale-up bioreactor" was tested in vitro for 18 days in static and flux conditions. Pig hepatocytes in the bioreactor were maintained in three-dimensional co-culture with non-parenchymal cells and are reorganised in a way similar to the liver cell plates in vivo: cells remained polarised in vitro clearly demonstrating biliary zones surrounding individual hepatocytes. The biochemical performance of the bioreactor was assessed by estimating its ability to remove two of the major toxins associated with hepatic encephalopathy: benzodiazepines and ammonia. The rates of ammonia elimination and drug biotransformation were maintained at constant high levels for almost two weeks. This "scaled-up bioreactor" provides conditions favourable for the formation of contiguous cell sheets, which allow to maintain constant liver specific functions. PMID- 11899897 TI - Highly efficient isolation of porcine islets of Langerhans for xenotransplantation: numbers, purity, yield and in vitro function. AB - Xenogeneic transplantation of porcine islets of Langerhans is regarded as a potential future treatment for diabetes mellitus. Despite considerable biotechnological progress, however, it is still very difficult and often unreliable to isolate sufficient numbers of highly purified, intact islets from the porcine pancreas with good in vitro function. OBJECTIVE: Of this study was to describe an efficient and reliable method to isolate sufficient numbers of highly purified islets of Langerhans with good in vitro function from adult as well as from young hybrid pigs. METHODS: Islets were isolated from the pancreas of young (4-6 months) hybrid pigs and old (2-3 years) retired breeders using Liberase PI and digestion-filtration. Average islet size was detected by dithizone staining of tissue sections prior to isolation; only organs with an average islet size > or = 200 microns were used. Density gradient purification with OptiPrep was performed in a COBE 2991 cell processor. Viability was investigated using fluorescence staining. Perifusion studies were carried out to asses in vitro function of isolated islets. RESULTS: Islets were successfully isolated from young hybrid pigs (3,671 +/- 598 IEQ/g) and old retired breeders (5,182 +/- 545 IEQ/g). After purification islet purity was 92% for retired breeders and 87% for young hybrid pigs. Yield after purification was still not satisfactory: 64% for retired breeders (3,209 +/- 444 IEQ/g) and 44% for young hybrid pigs (1,669 +/- 386 IEQ/g). Viability of isolated islets was 80-95%. Perifusion studies of porcine islets showed sufficient insulin release upon glucose challenge; however, the level of insulin release depended on the density of islets within the perifusion chamber. Low temperature culture (24 degrees C) prior to perifusion studies had no detrimental effect on insulin release. Long-term culture over 11 days was followed by a dramatic loss of islet function. CONCLUSIONS: If xenograft rejection can be overcome and the risk of xenosis can be minimised, sufficient numbers of purified porcine islets with good in vitro function can be isolated to serve as a potential source for islet transplantation in diabetic patients. PMID- 11899898 TI - Xenotransplantation: the good, the bad and the ugly. PMID- 11899899 TI - Current status and perspectives for the generation of transgenic pigs for xenotransplantation. AB - Xenotransplantation implies transplantation of organs between discordant, e.g. non-related species. This procedure usually is associated with a hyperacute rejection response (HAR) which destroys the transplanted organ within minutes. To overcome the growing shortage of human organs, transgenic pigs have been generated that express human complement regulatory genes. This approach enables to overcome the HAR as shown by an extended average survival rate of 40-90 days of the immunosuppressed primate recipient of a transgenic porcine heart. It is expected that transgenic pigs will be available as organ donors within the next 5 7 years. A major prerequisite is the prevention of the potential transfer of pathogenic microorganisms, in particular porcine endogenous retroviruses (PERV). Transgenic livestock has been generated predominantly via microinjection of DNA constructs into pronuclei of zygotes. However, efficiency is low and only 1-3% transgenic offspring are to be obtained. Integration of the transgene occurs at random and expression is independent from the number of integrated copies but can be affected by the integration site. Improvements of the efficiency in the generation of transgenic pigs will be achieved by the use of genetically modified donor cells in nuclear transfer technology (cloning). PMID- 11899900 TI - [The techniques of ultrasound diagnosis continue to progress very fast]. PMID- 11899901 TI - Comments on:non-associative fear acquisition: a review of the evidence from retrospective and longitudinal research. PMID- 11899902 TI - Current world literature. Erythrocytes. PMID- 11899903 TI - A randomized pilot study of SRL172 (Mycobacterium vaccae) in patients with small cell lung cancer (SCLC) treated with chemotherapy. AB - BACKGROUND: SRL172 is a suspension of heat killed Mycobacterium vaccae, that has been found to be a potent immunological adjuvant when used with autologous cells in animal models. This is a phase II study to test the clinical activity, feasibility and safety of combining SRL172 with chemotherapy to treat patients with small cell lung cancer (SCLC). METHODS: Patients were randomized to receive chemotherapy with (n=14) or without (n=14) SRL172. The chemotherapy was either platinum-based (MVP, n=10) or anthracycline-based (ACE, n=18). SRL172 was given intradermally on day 0, weeks 4, 8 and then 3-6 monthly. RESULTS: The treatment arms were well balanced for disease extent (43% with limited stage in each arm). The toxicity of chemotherapy and overall response at 12-15 weeks (57%) was the same for both treatment regimens. Median survival was 8.6 months and 12.9 for patients treated with chemotherapy alone and with the combination respectively (P=0.10). The survival trend was similar for both disease extent and chemotherapy regimen employed in favour of combination chemotherapy with SRL172. CONCLUSIONS: There is a trend to improved median survival in SCLC with the combination of chemotherapy and SRL172 with no increased toxicity and irrespective of drug regimen. A phase III study examining chemotherapy in combination with SRL172 in SCLC is now underway. PMID- 11899904 TI - Retrospective study of management of uterine sarcomas at Oxford 1990-1998: role of adjuvant treatment. AB - We report a retrospective study of 47 consecutive patients with uterine sarcoma treated at the Churchill Hospital in Oxford between 1990-1998. The mainstay of treatment was surgery with adjuvant chemotherapy and radiotherapy reserved for selected patients with early stage disease. Overall 1 and 2 year survival was 49% and 30% respectively compared with 73% and 55% in the group who received adjuvant chemotherapy/radiotherapy. Median survival was 11 months for the group as a whole compared to 32.9 months in the adjuvant therapy group. This is a retrospective review with small numbers and considerable selection bias, however, given the poor survival of patients with this disease, adjuvant treatment should be considered in future trials of patients with uterine sarcoma. PMID- 11899905 TI - Short report: a morbidity scoring system for Clinical Oncology practice: questionnaires produced from the LENT SOMA scoring system. PMID- 11899906 TI - Primary treatment endpoint following palliative radiotherapy for painful bone metastases: need for a consensus definition? AB - OBJECTIVES: To compare and contrast the definitions of primary treatment endpoints in randomized studies of dose-fractionation schedules for treating bone metastases and to identify basic characteristics of treatment endpoint that may require consensus among investigators. METHODS: Randomized controlled trials (RCTs) of various dose-fractionation schedules for painful bone metastases, published between 1980 and 1999, and on-going trials whose protocols were available, were systematically reviewed based on the following features of the primary treatment endpoint: (i) degree of pain relief; (ii) timing of the pain response assessment; (iii) effect of co-interventions on pain relief; (iv) the reduction of analgesic as a treatment response; and (v) quantification of response duration. RESULTS: Ten published RCTs (each sampled over 100 patients), plus two current trial protocols were reviewed. Five of the 12 studies defined any reduction in pain score as the primary endpoint. Three trials defined response at pre-determined time points, whereas eight studies attributed pain improvement at any time during follow-up to the effect of radiotherapy. No trial incorporated effect of systemic treatments on response. Only two trials incorporated analgesic scores into the primary endpoint criteria, although several trials reported results of combined pain and analgesic relief. Eight trials reported duration of response. Three provided some estimation of duration with respect to survival: two of them employing actuarial time to pain progression, and one calculated the ratio of pain response to median survival duration (percent net relief). Quality of life was measured in four of 12 studies, as secondary endpoint. CONCLUSION: Although available data suggest similarity in pain relief among various dose-fractionation schedules, accurate and consistent description of the degree of benefit from radiotherapy is lacking. While pain relief is a consistent primary treatment goal among randomized trials, a consensus on several important features of treatment endpoint is needed in order to establish common grounds for future trials in palliative radiotherapy. PMID- 11899907 TI - The role of daily activities in pressure ulcer development. PMID- 11899908 TI - Vasculitis: a precis. PMID- 11899910 TI - Home health wound services under Medicare PPS. PMID- 11899909 TI - Tube feeding and pressure ulcers. PMID- 11899911 TI - Documentation with MDS Section M: Skin Condition. AB - The Minimum Data Set is designed to be the assessment instrument used in all long term-care facilities receiving federal funds for Medicare and Medicaid. However, Section M: Skin Condition is one of the most challenging sections to complete when trying to match the Minimum Data Set documentation with the true clinical picture. If wounds are not adequately assessed and documented, outcomes of care cannot be evaluated, and treatment and prevention plans will be inadequate. This may result in less than optimal outcomes and possible lawsuits for inadequate care against both the caregiver and facility. The purpose of this article is to provide examples of medical record documentation necessary to support the Minimum Data Set assessment. PMID- 11899912 TI - TGF-beta3 in the treatment of pressure ulcers: a preliminary report. AB - OBJECTIVE: To determine the safety, tolerability, and efficacy of TGF-beta3 in the treatment of chronic, nonhealing pressure ulcers. DESIGN: A subset analysis of data from a randomized, blind, parallel, placebo-controlled trial involving 270 patients. SETTING: University of Michigan Wound Care Center. PATIENTS: A total of 14 patients (6 women and 8 men aged > or = 18 years) with pressure ulcers were randomly assigned to 1 of 3 groups to receive once daily topical application of recombinant TGF-beta3 or placebo gel for a period of no more than 16 weeks. Group 1 (n=4) received 1.0 microg/cm2 of TGF-beta3, Group 2 (n=5) received 2.5 microg/cm2 of TGF-beta3, and Group 3 (n=5) received placebo gel. All subjects received standardized wound care as well. Weekly evaluations were performed for efficacy, determined by relative wound surface areas and volumes, and were compared with initial baseline values and safety parameters. MAIN OUTCOME MEASURE: Reduction in pressure ulcer area and volume. MAIN RESULTS: Group 2 had a significantly increased rate of wound healing at the fourth visit (P<.05). No significant difference was observed in the healing rate among the groups at the termination of the study. Treatment with TGF-beta3 was well tolerated and there were no significant adverse reactions. CONCLUSION: The findings of this study indicate that the topical application of TGF-beta3 is safe and useful in the treatment of pressure ulcers and is most effective at the earliest stages of therapy. PMID- 11899913 TI - Options for nonsurgical debridement of necrotic wounds. PMID- 11899914 TI - Comparative genomics by capture PCR. AB - There is increasing demand for efficient methods to relate genomic information from model organisms to other species of interest. Comparative genetic analyses are particularly valuable to identify functionally important sequence features on the basis of their evolutionary conservation. We demonstrate here how a single segment of just 32 or less conserved coding nucleotide positions can be used to isolate homologous gene sequences from large numbers of species using a single sided PCR technique. The method was used to isolate and determine the 3' untranslated sequence of the somatostatin gene from vertebrate species ranging from human to hagfish. Two sequence motifs centered an average 40-145 nucleotides downstream of the translational stop codon have remained conserved for up to 350 million years. One of the conserved tetrapod segments was used to select a primer for amplification of so-called comparative anchor tagged sequences (CATS) in regular PCR, and shown to amplify homologous sequences from DNA samples from 30 out of 33 tetrapods. In conclusion, we present a useful procedure to reveal functionally relevant sequence elements, and to select primers for amplification of homologous sequences from a wide range of species. PMID- 11899915 TI - Training in anesthesia for rural family physicians.. PMID- 11899916 TI - [Criteria for Plavix supplementation]. PMID- 11899917 TI - Overt honesty measures predicting admissions: an index of validity or reliability. AB - Honesty questionnaires are often validated against a paper-and-pencil criterion in which respondents are asked to admit to past incidents of dishonesty. However, substantial overlap in the methods of assessment and in item content between predictor and criterion suggest that it is not validity that is being assessed, but rather a form of reliability. In this study, the relations between two overt measures of honesty (the Phase II Profile and the Workplace Productivity Questionnaire) and an admissions criterion were evaluated. The results suggest that the strong correlations between the Phase II Profile and the admissions criterion (r = -.67) and between scores on the Workplace Productivity Questionnaire and the admissions criterion (r = -.62) may be largely due to overlapping item content. PMID- 11899918 TI - Aspects of the construct validity of a preliminary self-concept questionnaire. AB - The aim of the study was to assess the construct validity of a Preliminary Self concept Questionnaire and an Afrikaans version of the Self-description Questionnaire of Marsh, Smith, and Barnes. Questionnaires were administered to 57 boys and 57 girls in Grades 5 and 7. Their self-concepts were rated by their teachers on a teacher's rating scale, based on the four subscales of the Preliminary Self-concept Questionnaire. Correlations among the total scores of the three self-concept measures were calculated, as well as correlations among corresponding subscale scores. These values supported the construct validity of the Preliminary Self-concept Questionnaire and the convergent validity of three of the four subscales. PMID- 11899919 TI - Confirmatory factor analysis and job burnout correlates of the Health Professions Stress Inventory. AB - Previous research in 1994 by Gupchup and Wolfgang identified four factors from Wolfgang's Health Professions Stress Inventory (1988) that were common among a sample of practicing pharmacists. The factors were labeled Professional Recognition. Patient Care Responsibilities, Job Conflicts, and Professional Uncertainty, respectively. We used confirmatory factor analysis to assess whether this factor structure was generalizable to nurses. To examine concurrent validity, we correlated the factors with Maslach and Jackson's three dimensions of job burnout, i.e., Emotional Exhaustion, Depersonalization, and Personal Accomplishment. Data were collected through a questionnaire survey of a random sample of 9,380 nurses from across 43 public hospitals in Hong Kong, from which 2,267 (24.2%) responded. Analysis indicated statistically acceptable goodness of fit indices for the four-factor solution. Except for the factor Patient Care Responsibilities. all other factors had moderate correlations between .44 and .53 with Emotional Exhaustion and Depersonalization. Correlations between the factors of Stress Inventory and Personal Accomplishment were small but significant, ranging from -.25 to .13. Areas for further improving the psychometric properties of the inventory are discussed. PMID- 11899920 TI - Relation of study strategies to the academic performance of Hong Kong university students. AB - A revised version of the Learning and Study Strategies Inventory was used to examine the relation of study strategies with academic performance of 100 Hong Kong university students. Analysis indicated the high academic-achieving group differed significantly from the low academic-achieving group in terms of intrinsic disposition factors of motivation, scheduling, concentration, and selecting main ideas. PMID- 11899921 TI - Recognition and diagnosis of lysosomal storage diseases in the cat and dog. AB - Lysosomal storage diseases are rare, inherited disorders caused by the deficiency of 1 or more enzymes within the lysosomes of cells or by the deficiency of an activating protein or cofactor necessary for enzyme activity. The enzyme deficiency leads to a catabolic blockade and subsequent accumulation of storage material, and this in turn leads, albeit indirectly, to a wide array of clinical signs. Many features of storage diseases make them difficult to recognize and diagnose. In this review, we summarize the clinical features of these diseases and outline the steps required to confirm a diagnosis. PMID- 11899922 TI - A prospective study on active and environmental tobacco smoking and bladder cancer risk (The Netherlands). AB - OBJECTIVE: In a prospective cohort study among 120,852 adult subjects the authors investigated the associations between cigarette, cigar, pipe, environmental tobacco smoking (ETS), and bladder cancer. METHODS: In 1986 all subjects completed a questionnaire on cancer risk factors. Follow-up for incident bladder cancer was established by linkage to cancer registries until 1992. The case cohort analysis was based on 619 cases and 3346 subcohort members. RESULTS: Compared with lifelong non-smokers the age- and sex-adjusted incidence rate ratios (RR) for ex- and current cigarette smokers were 2.1 (95% CI 1.5-3.0) and 3.3 (95% CI 2.4-4.6), respectively. The RR for smoking duration was 1.03 (95% CI: 1.02-1.04) per 1-year increment. The RR per 10 cigarettes/day was 1.3 (95% CI 1.2 1.4). Tar and nicotine exposure increased bladder cancer risk only weakly. It appeared that associations of cigarette smoking characteristics with bladder cancer risk were largely attributable to cigarette smoking duration only. Smoking cessation, age at first exposure, filter-tip usage, cigar and pipe smoking, and ETS were no longer associated with bladder cancer risk after adjustment for frequency and duration of smoking. CONCLUSIONS: The authors conclude that current cigarette smokers have a three-fold higher bladder cancer risk than non-smokers. Ex-smokers experience a two-fold increased risk. About half of male bladder cancer and one-fifth of female bladder cancer was attributable to cigarette smoking. Other smoking types (cigar, pipe, or ETS) were not associated with increased risks. PMID- 11899924 TI - [Newsletter and archives of the Lombard Association of Occupational Health Physicians. 1946]. PMID- 11899923 TI - [Aplastic myelosis and subsequent leukopenic leukemia, caused by benzene. 1945]. PMID- 11899926 TI - [The Occupational Health Clinic in safeguarding the health printing industry workers. 1948]. PMID- 11899925 TI - [Reflections on benzene-induced leukemia. 1948]. PMID- 11899927 TI - [Unusual epidemics of mercury poisoning in a felt hat factory. 1949]. PMID- 11899928 TI - [Contribution to the anatomopathologic study of pulmonary asbestosis. 1949]. PMID- 11899929 TI - [Changes in the central nervous system of vascular origin in carbon disulphide poisoning. 1950]. PMID- 11899930 TI - [The Debye-Scherrer method in the analysis of silicon in industrial dust. 1951]. PMID- 11899931 TI - [Broncho-alveolar structure and topography of pneumoconiotic lesions. 1951]. PMID- 11899932 TI - [Beryllium-induced granulomatosis. 1951]. PMID- 11899933 TI - [Unemployment and morbidity. Observations on a group of 1,120 unemployed individuals hired at work. 1954]. PMID- 11899934 TI - [Asbestosis and pulmonary carcinoma in a female asbestos fibre spinner (remarks on the cancer problem of asbestos). 1955]. PMID- 11899935 TI - [Methods for analysis of free crystalline silica]. PMID- 11899936 TI - [The immunologic etiology of silicosis. Part Ia: General considerations. 1958]. PMID- 11899937 TI - [The immunologic etiology of silicosis. Part IIa: Biochemical and immunologic considerations on hyaline. 1958]. PMID- 11899938 TI - [Modern trends in the treatment of severe respiratory insufficiency in chronic pneumopathies. 1958]. PMID- 11899939 TI - The histogenesis of experimental silicosis. I--Methods for the histological evaluation of experimentally induced dust lesions. (2). 1960. PMID- 11899940 TI - Counting of atmospheric dusts using millipore filters. 1960. PMID- 11899942 TI - [Assessment of intensity of work performed during heavy vehicle driving with power and mechanical steering. 1965]. PMID- 11899943 TI - [Erythrocyte ala-dehydratase activity as a test of occupational lead poisoning. 1966]. PMID- 11899941 TI - Rheumatoid pneumoconiosis syndrome. 1965. PMID- 11899944 TI - [Significance and limits of respiratory function tests in the study of byssinosis. Study at a cotton factory in Lombardia. 1967]. PMID- 11899945 TI - [Pathologic anatomy of silicosis. 1967]. PMID- 11899946 TI - [Changes in isoenzymatic fractions of erythrocyte lactate-dehydrogenase (LDH) in lead poisoning anemia. 1968]. PMID- 11899947 TI - Pathology of vinyl chloride. 1970. PMID- 11899948 TI - Effect of a standardized meal on the bioavailability of a single oral dose of tibolone 2.5 mg in healthy postmenopausal women. AB - STUDY OBJECTIVE: To assess the effect of food on absorption and pharmacokinetic disposition of tibolone. DESIGN: Open-label, randomized, crossover study with a 1 week washout period. SETTING: Institut fur Klinische Pharmakologie, Hohenkirchen Siegertsbrunn, Germany SUBJECTS: Twenty-four healthy, early postmenopausal women. INTERVENTION: Single doses of tibolone 2.5 mg were administered after subjects consumed a high-fat meal or fasted. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of tibolone and its three primary metabolites, delta4-tibolone, 3alpha-hydroxytibolone, and 3beta-hydroxytibolone, were assayed by gas chromatography with mass spectrometry. Peak plasma concentration (Cmax), time to Cmax, area under the plasma concentration versus time curve from time zero to infinity (AUC(0-infinity)), and elimination half-life were calculated, and food effects were evaluated. Plasma concentrations of tibolone and delta4-tibolone were too low to estimate AUC(0-infinity) and half-life. Absorption or formation of 3alpha-hydroxytibolone and 3beta-hydroxytibolone was slower in fed participants than in fasting participants, but this was of no clinical relevance because of the long-term nature of tibolone treatment. Meal consumption did not affect AUC(0-infinity) or half-life for 3alpha-hydroxytibolone and 3beta hydroxytibolone. CONCLUSION: Food consumption decreased and delayed Cmax but had no effect on the exposure of tibolone and its metabolites. Tibolone therefore can be administered irrespective of meal timing. PMID- 11899949 TI - Role of fomepizole in the management of ethylene glycol toxicity. AB - OBJECTIVE: To systematically review English-language articles on fomepizole administration in patients with ethylene glycol poisoning. DATA SOURCES: MEDLINE, EMBASE, Current Contents, and PubMed. Search terms were fomepizole, 4 methylpyrazole, and ethylene glycol. The search was supplemented with a bibliographic review of all relevant articles. STUDY SELECTION: All published reports of fomepizole administration in patients with ethylene glycol poisoning were reviewed, irrespective of study design. We identified one clinical trial and subsequent pharmacokinetic study, one case series, and 13 case reports. RESULTS: Fomepizole has been investigated in 70 patients in open, unblinded studies. Most patients received an intravenous loading dose, with subsequent variable maintenance doses every 12 hours until plasma ethylene glycol levels became undetectable. Additional hemodialysis treatment generally was administered when patients had renal insufficiency or ethylene glycol levels above 50 mg/dl. Many patients had detectable ethanol levels either because of coadministration or as a result of adjunctive treatment at a referring center. Poorer patient outcomes, such as death and renal insufficiency, were associated with later clinical presentation time after ingestion. At therapeutic fomepizole levels (> 8.6 mg/ml), the half-life of ethylene glycol was prolonged to over 19 hours. Fomepizole appeared to be well tolerated by most patients. CONCLUSION: Fomepizole is an effective alcohol dehydrogenase inhibitor that decreases production of ethylene glycol metabolites. Reduced mortality and morbidity are undetermined because of the small number of patients evaluated to date. Data on comparative efficacy of fomepizole versus ethanol and data on administration of fomepizole in children are limited. PMID- 11899950 TI - Laboratory-scale preparation of soft cheese artificially contaminated with low levels of Escherichia coli O157, Listeria monocytogenes, and Salmonella enterica serovars Typhimurium, Enteritidis, and Dublin. AB - The production of cheese with incurred low levels of pathogenic microorganisms stressed by the production process was the aim of the study. A standard protocol for the preparation of artificially contaminated soft cheese on a laboratory scale was developed. Milk for cheese preparation was artificially contaminated with pathogenic target microorganisms at low levels, between 1 and 10 CFU/ml. Two strains of Escherichia coli OI157:H7, two strains of Listeria monocytogenes, and three Salmonella spp. (Salmonella enterica serovars Typhimurium, Enteritidis, and Dublin) were investigated. The food pathogens in the cheese were exposed to the entire production process. All three microorganism species survived the cheese production process and were detected in the final product at concentrations between 1 and 50 CFU/g. The cheese produced contains target microorganisms that have been exposed to curd formation, drainage, setting, and ripening. This cheese can be used to validate microbiological methods or to examine the target microorganisms in a natural food environment at low concentrations. It represents an alternative to artificial contamination of cheese by adding target microorganisms to a final cheese product. PMID- 11899951 TI - Recovery of Listeria monocytogenes from vacuum-sealed packages of frankfurters: comparison of the U.S. Department of Agriculture (USDA) food safety and inspection service product composite enrichment method, the USDA Agricultural Research Service (ARS) product composite rinse method, and the USDA-ARS package rinse method. AB - This study compared three methods for the recovery of Listeria monocytogenes from commercially prepared and vacuum-packaged frankfurters that were inoculated with a five-strain mixture of this pathogen at averages of 22 and 20,133 CFU per package over three trials. The presence and levels of the pathogen were determined by (i) the U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS) product composite enrichment method, involving the selective enrichment of a 25-g composite of product and the subsequent plating of this product onto selective agar plates; (ii) the USDA Agricultural Research Service (ARS) product composite rinse method, involving the rinsing of a 25-g composite of product with 0.1% peptone water and the subsequent plating of a portion of the rinse fluid directly onto selective agar plates; and (iii) the USDA-ARS package rinse method, involving the use of 25 ml of 0.1% peptone water to rinse the entire contents of a package and the subsequent plating of a portion of the rinse fluid directly onto selective agar plates. For packages inoculated with 20,133 CFU. L. monocytogenes was recovered at a frequency (percentage of packages positive) of 100% by each of the three methods. The pathogen was recovered at efficiencies (percentages of recovery of L. monocytogenes) of 43 and 94% with the USDA-ARS product rinse method and the USDA-ARS package rinse method, respectively. For packages inoculated with 22 CFU, L. monocytogenes was recovered at frequencies of 17, 10, and 100% by the USDA-FSIS product composite enrichment method, the USDA-ARS product composite rinse method, and the USDA-ARS package rinse method, respectively. The pathogen was recovered at efficiencies of 20 and 95% with the USDA-ARS product composite rinse method and the USDA-ARS package rinse method, respectively. In a related study, the USDA-ARS package rinse method was the only method that detected the pathogen in 60 packages from each of five brands of frankfurters purchased from local grocery stores. These data establish that the USDA-ARS package rinse method is markedly more sensitive, as well as demonstrably more rapid and facile, than either the approved USDA-FSIS product composite enrichment method or the USDA-ARS product composite rinse method in determining the presence or absence of L. monocytogenes and establishing the levels of the pathogen that may be on the surface of ready-to-eat foods such as frankfurters. PMID- 11899952 TI - Vitamin D and vitamin D analogs in cancer treatment. AB - The secosteroid hormone 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) is a key player in the regulation of bone mineralization and calcium homeostasis. In addition, 1,25-(OH)2D3 has antiproliferative and prodifferentiation effects on various cells in vitro and in vivo. The growth-inhibitory properties of 1,25-(OH)2D3 could be harnessed in the treatment of cancer. However, its use as an anti-cancer drug is limited because of the calcemic effects of pharmacological doses. In an attempt to dissociate the antiproliferative and calcemic effects, numerous vitamin D3 analogs were developed. The mechanisms by which 1,25-(OH)2D3 and 1,25 (OH)2D3 analogs exert their growth-inhibitory effects are not clear but include effects on cell differentiation, apoptosis, cell cycle regulation, metastases, and angiogenesis. In the current review aspects involved in the tumor suppressive activity of 1,25-(OH)2D3 and 1,25-(OH)2D3 analogs will be addressed. The use of vitamin D3 compounds, alone or in combination with other drugs, in cancer treatment and the potential drawbacks will also be discussed. PMID- 11899953 TI - Scleroderma-like nailfold capillaroscopic abnormalities are associated with anti U1-RNP antibodies and Raynaud's phenomenon in SLE patients. AB - This study aimed to evaluate the association between nailfold capillary abnormalities and the presence of Raynaud's phenomenon (RP), anti-U1-RNP, and anti-cardiolipin (aCL) antibodies in SLE patients. One-hundred SLE patients were studied. Widefield nailfold capillaroscopy was considered abnormal according to five criteria. Intercapillary distance, capillary width and capillary length were registered by videomorphometry in two fingers in 100 patients and in four fingers in 40 of these patients. Both the presence of alterated capillaroscopy and the presence of scleroderma-pattern (SD-pattern), characterized by the presence of avascular areas and enlarged or giant loops, were associated with the isolated presence of RP (P < 0.001) or anti-U1-RNP antibodies (P < 0.01), as well as with the simultaneous presence of RP and anti-U1RNP antibodies (P < 0.001). There was a negative association between the presence of aCL antibodies and SD-pattern (P < 0.05). Higher figures for the videomorphometric parameters capillary width, intercapillary distance and capillary length (measured on four fingers) were observed in patients with RP. Patients presenting both RP and anti-U1-RNP antibodies showed higher figures for intercapillary distance and capillary width. This study demonstrated significant association between nailfold capillaroscopic abnormalities and either RP or anti-U1-RNP antibodies in SLE patients. The association of RP, anti-U1-RNP antibodies, and 'scleroderma-like' findings on nailfold capillaroscopy (SD-pattern) in patients with SLE may suggest a new SLE subset with subclinical features of systemic sclerosis. PMID- 11899954 TI - Multimodality therapy for resectable cancer of the thoracic esophagus. AB - The frequency of esophageal carcinoma continues to increase in North America primarily because of the increased incidence of Barrett's epithelium in the distal esophagus and its malignant potential. Aggressive treatments involving multimodality therapies have been offered to improve overall poor survival rates. A review of this experience follows, to explain the rationale and to compare results of therapies. Although preoperative chemoradiation therapy is commonly used for locally advanced disease, few data support its superiority over surgical resection alone, followed by adjuvant therapy when appropriate. Hence this regimen should be limited to patients enrolled in controlled, randomized studies until the data support its widespread use. PMID- 11899955 TI - Simple pulmonary vein isolation for atrial fibrillation. PMID- 11899956 TI - Paravalvular leak and other events in silzone-coated mechanical heart valves: a report from AVERT. AB - BACKGROUND: The Artificial Valve Endocarditis Reduction Trial (AVERT) was designed to compare endocarditis rates in Silzone versus conventional valves. Recruitment ended January 21, 2000, because of higher rates of paravalvular leakage in patients receiving the Silzone prosthesis. The present analysis determined late event rates that might be used in the management of approximately 36,000 patients who have received the Silzone prosthesis. METHODS: A total of 807 patients in 19 centers in North America and Europe were randomized. Mean age was 61+/-11 years; 41% were women. Operations included aortic valve replacement in 59%, mitral valve replacement in 32%, and aortic and mitral valve replacements in 9%; 41% had concomitant operations (26% coronary artery bypass grafting). RESULTS: Major paravalvular leakage (followed by repair, explant, or mortality) occurred in 18 of 403 patients receiving Silzone valves and 4 of 404 patients without Silzone valves (2-year event-free rates: 91.1% versus 98.9% conventional, p < 0.003). Similarly, 2-year freedom from any explant was lower in the Silzone arm (19 versus 2 events; 90.1% versus 99.4%, p = 0.0002). Rates of mortality and stroke were similar during follow-up. CONCLUSIONS: Continued follow-up of AVERT supports the conclusion that the Silzone prosthesis has increased risk of paravalvular leakage requiring reoperation. Overall survival is similar in the two groups. PMID- 11899957 TI - Epicardial ultrasound in off-pump coronary artery bypass grafting: potential aid in intraoperative coronary diagnostics. AB - BACKGROUND: In off-pump coronary artery bypass surgery (OPCAB), epicardial ultrasound may aid in several intraoperative dilemmas. The aim of this study was to test a new mini-transducer for intraoperative coronary diagnostics. METHODS: A 10 MHz mini-transducer (15 x 6 x 9 mm) was applied epicardially in eight open chest and two closed chest porcine OPCAB procedures (using robotics) and on four postmortem human hearts. The transducer fitted in between the suction pods of the cardiac stabilizer and passed an 11-mm port. RESULTS: In the open chest cases the internal mammary arteries (including the side branches) could be visualized totally (n = 12). The left anterior descending coronary artery could be located over its entire course. Vascular anatomy, side branches, and septal perforators (diameter > or = 0.2 mm) could easily be discerned. In the closed chest cases the left anterior descending coronary artery, its side branches, and septal perforators could be visualized in both cases. In the postmortem human hearts the left anterior descending coronary artery could be visualized totally under the thick epicardial fibro-fatty layer and pathologic conditions could be identified. CONCLUSIONS: The 10 MHz ultrasound mini-transducer showed promise as a diagnostic tool in both open and closed chest coronary procedures on the beating heart. PMID- 11899958 TI - Vascular endothelial growth factor-mediated, endothelium-dependent relaxation in human internal mammary artery. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) has been shown to have potential to treat ischemic diseases. Moreover, its vasorelaxing or vasodilatory effect might be favorable for relieving graft spasm. In this study, we examined the vasorelaxing effects of recombinant VEGF in isolated human internal mammary artery (IMA) and compared the responses to acetylcholine and nitroglycerin. METHODS: Isometric tension of IMA ring segments was measured with an organ bath technique. With an optimal resting tension determined from its individual length tension curve, precontraction was induced by 10(-8) M U46619 and cumulative concentration-relaxation was measured by application of VEGF (10(-12) to 10(-15) M), acetylcholine (10(-10) to 10(-5) M), and then nitroglycerin (10(-4.5) M). RESULTS: Vascular endothelial growth factor induced concentration-dependent relaxation (EC50: -9.89+/-0.05 log M; Emax: 63.2%+/-7.3%) in IMA with intact endothelium. The relaxant responses to VEGF were significantly attenuated by pretreatment with Nomega-nitro-L-arginine (L-NNA) alone and indomethacin + L-NNA, and totally abolished by removal of the endothelium or pretreatment with indomethacin + L-NNA + oxyhemoglobin. Internal mammary arteries became more sensitive to VEGF in the presence of indomethacin alone. However, acetylcholine induced relaxation was not abolished by treatment with indomethacin + L-NNA + oxyhemoglobin (Emax: 16.9%+/-2.7%). The endothelium-independent relaxations induced by nitroglycerin were also significantly inhibited by administration of oxyhemoglobin. CONCLUSIONS: The results demonstrate that VEGF-induced endothelium dependent relaxation in the human IMA is mainly due to nitric oxide release. Although the vasorelaxing effect is not the primary advantage of this drug when it is used for angiogenesis, such effect may be advantageous in patients who also need a coronary artery bypass operation. PMID- 11899959 TI - Successful repair of intraoperative aortic dissection detected by transesophageal echocardiography. AB - Aortic dissection is a rare but devastating complication of cardiac surgery. Early intraoperative diagnosis and management are essential for a favorable outcome. We describe the case of a 69-year-old man with worsening dyspnea who was admitted for mitral valve replacement having previously had a mitral valve repair. Precardiopulmonary bypass transesophageal echocardiography confirmed mitral regurgitation and showed mild atherosclerotic changes in the descending aorta. Following successful replacement of the mitral valve, an attempt to wean from cardiopulmonary bypass failed. This was characterized by acute onset hypovolemia. The transesophageal echocardiography showed the presence of features of acute aortic dissection involving only the descending aorta without identifying the entry point. The tear was successfully repaired by direct suture within the lumen. PMID- 11899960 TI - Right coronary artery and interatrial septal aneurysms with fistulous connection to the right atrium. AB - Coronary artery aneurysmal disease is a rare pathology occasionally associated with coronary artery to venous fistulous connection. We report a case of right coronary artery aneurysm with fistulous connection to the right atrium associated with an aneurysm of the interatrial septum. PMID- 11899961 TI - Left atrial cardiac hemangioma associated with shortness of breath and palpitations. AB - We present a patient with a left atrial hemangioma associated with shortness of breath and irregular heart-beats. Imaging modalities used to evaluate this rare tumor demonstrated characteristic features of its vascular nature. After successful surgical excision of the tumor under cardiopulmonary bypass, there were no clinical or echocardiographic evidence of recurrence at 18 months. PMID- 11899962 TI - Left ventricular assist device implantation via left thoracotomy: alternative to repeat sternotomy. AB - Repeat sternotomy for left ventricular assist device insertion may result in injury to the right heart or patent coronary grafts, complicating intraoperative and postoperative management. In 4 critically ill patients, left thoracotomy was used as an alternative to repeat sternotomy. Anastomosis of the outflow conduit to the descending thoracic aorta provided satisfactory hemodynamic support. PMID- 11899963 TI - The emergence and application of technological advances in biotransformation studies. AB - The past years have seen only the beginning of our understanding of metabolic processes and the importance of these processes to the development of safe and effective medicines. The trend to bring more detailed information into earlier stages of drug discovery will continue to drive improvements in technology and in experimental and analytical procedures for the study of biotransformation of drugs. The challenges are significant, but so is the promise of the contributions that can be made by biotransformation studies. PMID- 11899964 TI - Beneficial effects of chromium in people with type 2 diabetes, and urinary chromium response to glucose load as a possible indicator of status. AB - No reliable method for the estimation of chromium (Cr) status is available yet. The aim of this study is to investigate the possibility of using urinary Cr response to glucose load as an indicator of Cr status. Seventy-eight non-insulin dependent diabetes mellitus patients, were divided randomly into two groups and given Cr supplements as brewer's yeast and CrCl3 sequentially with placebo in between, in a double-blind, crossover design of four stages, each lasting 8 wk. At the beginning and end of each stage, subjects were weighed, their dietary data and drug dosage recorded, and blood and urine samples collected for analysis of glucose and urinary chromium (fasting and 2 h post-75-g glucose load) and fructosamine. The mean urinary Cr after the glucose load was significantly higher than the fasting mean at zero time (p<0.01). However, only 52 of the patients showed an obvious increase; the others showed a slight decrease or no change. Both supplements caused a significant increase in the means of urinary Cr and a significant decrease in the means of glucose and fructosamine. Only those subjects responding to Cr supplement by improved glucose control showed an increase in post-glucose-load urinary Cr over fasting level, after the supplement but not at zero time. Therefore, it was concluded that urinary Cr response to glucose load could be used as an indicator of Cr status. PMID- 11899965 TI - The "redisorganisation" of the NHS. Radical changes can be made only if the basic environment is stable. PMID- 11899966 TI - The "redisorganisation" of the NHS. Doctors must have more managerial training. PMID- 11899967 TI - The "redisorganisation" of the NHS. Doctors working in public health have extra worries in latest reorganisation. PMID- 11899968 TI - The "redisorganisation" of the NHS. GPs' dissatisfaction with NHS plan is worse than editorial suggests. PMID- 11899969 TI - Effectiveness of guidelines on persistent glue ear in children. Effect claimed may depend on how persistent glue ear is defined. PMID- 11899970 TI - Effectiveness of guidelines on persistent glue ear in children. Guidelines in Australia were less effective than guidelines in England. PMID- 11899971 TI - Why general practitioners do not implement evidence. Learning environments must be created that capitalise on teams' wealth of knowledge. PMID- 11899972 TI - Adult obesity and growth in childhood. Obstetricians seem reluctant to consider interventions to reduce mean birth weight. PMID- 11899973 TI - Adult obesity and growth in childhood. Association of birth weight with adult weight is confounded by maternal body mass index. PMID- 11899974 TI - Adult obesity and growth in childhood. Mothers tend to pass their dietary habits on to their children. PMID- 11899975 TI - Adult obesity and growth in childhood. Factors that programme resistance to obesity must be identified. PMID- 11899976 TI - End-of-life care models highlighted in report. PMID- 11899977 TI - Simple clinical tests help determine older women's risk for early physical disability. PMID- 11899978 TI - Statement outlines competencies needed for cardiac stress testing. PMID- 11899979 TI - Coordinated anticoagulation service improves therapy outcomes and reduces monitoring costs. PMID- 11899980 TI - Modern imaging methods and preoperative management of pheochromocytoma: review of the literature and case report. AB - OBJECTIVES: To discuss the modern imaging techniques and preoperative management of pheochromocytoma and to report on one additional case. METHODS: A 66-year-old male with an incidentally discovered left adrenal mass is described. The adrenal medulla strongly accumulate 131 I-metaiodobenzylguanidine (MIBG). The patient underwent left adrenalectomy after preoperative therapy with alpha and beta blockers. The recent literature on pheochromocytoma modern imaging techniques and preoperative management is reviewed. RESULTS: MIBG scintigraphy diagnosed a benign functioning adrenal pheochromocytoma, allowing preoperative medical management. Postoperative workup was unremarkable. Diagnosis of pheochromocytoma was confirmed by immunohistopathology. At 18 months follow-up, the patient is alive and disease-free. CONCLUSIONS: Incidentally discovered adrenal masses have to be investigated to detect malignancy and subtle hormonal overproduction. MIBG scintigraphy has a high specificity (100%) in detecting pheochromocytoma, metastasis, surgical residual tumor, local relapse and other adrenal crest tumors. Positive results of octreotide scintigraphy in detecting malignant pheochromocytoma have been reported. Currently, pheochromocytoma removal is a safe operation with mortality rates ranging from 0 to less than 3%. Preoperative alpha adrenergic blockage with phenoxybenzamine or prazosin is important in decreasing the operative risk. Beta-blockers may be necessary for cardiac arrhythmia. Intraoperative invasive monitoring of hemodynamic variables may be both diagnostic and therapeutic of inadequate preoperative management. Lifelong follow-up for patients with pheochromocytoma is important. PMID- 11899982 TI - Coordinated pharmaceutical care assessment program improves clinical outcomes for high-risk elderly patients. PMID- 11899981 TI - New guidelines are a comprehensive resource for managing infection risk among hematopoietic stem cell transplant patients. PMID- 11899983 TI - Treat children's chronic pain as distinct from adults', statement says. PMID- 11899984 TI - Organizations take aim at medical mistakes, get honors. PMID- 11899985 TI - Should we be concerned about the content of mission statements for Christian hospitals? PMID- 11899986 TI - Christian credentials for Roman Catholic health care: medicine versus the healing mission of the church. PMID- 11899987 TI - Catholic hospitals and Catholic identity. AB - Catholic hospitals seek to offer health care in accord with the example of Christ. They have several models to assist in this effort. The first model is the values portrayed in the Gospels. The Catholic Church has sought to embody these Gospel values in specific teachings. These teachings have been further specified for hospitals in the United States by the National Conference of Catholic Bishops in the Ethical and Religious Directives. Finally, the Gospels values are also expressed for individual Catholic health care systems in mission statements and statements of Catholic identity. This article examines the worth of mission and identity statements, and explains that the statements must be put into practice through a process of internalization before they will be able to be of worth to the Catholic health care apostolate. PMID- 11899988 TI - The deChristianization of Christian health care institutions, or, how the pursuit of social justice and excellence can obscure the pursuit of holiness. PMID- 11899989 TI - Catholic health care institutions: dinosaurs awaiting extinction or safe refuge in a culture of death. PMID- 11899990 TI - Roman Catholic health care identity and mission: does Jesus language matter? AB - This article examines the current use of Jesus language in a convenience sample of twenty-five mission statements from Roman Catholic hospitals and health care systems in the United States. Only twelve statements specifically use the words "Jesus" or "Christ" in their mission statements. The author advocates the use of explicit Jesus language and modeling. While the witness of Jesus in the Gospel healing narratives is not only the corrective to current abuses in the health care delivery system, it is foundational to the integrity of Roman Catholic health care identity and mission. An analysis of Gospel healing narratives is used to illustrate the prophetic dimension of Jesus' wisdom, word, and witness. PMID- 11899991 TI - Institutional identity and Roman Catholic hospitals. PMID- 11899992 TI - Living the vision: health care, social justice and institutional identity. AB - This paper will examine the topic of identity in Roman Catholicism from the perspective of topics contained in or absent from mission statements of 25 Catholic health care institutions. In particular, I will look at these from the perspective of social justice as well as how this and other topics such as human dignity, sanctity of life, stewardship, pastoral care and the likelihood of mergers with other institutions will affect the healing ministry of Catholic health care providers. The article will conclude that there are three key dimensions to Catholic health care: leadership in advocating reform of the current health care system, care for the marginalized and under-insured, and the provision of pastoral care in all institutions. PMID- 11899993 TI - Organizational ethics in Catholic health care: honoring stewardship and the work environment. AB - Organizational ethics refers to the integration of values into decision making, policies, and behavior throughout the multi-disciplinary environment of a health care organization. Based upon Catholic social ethics, stewardship is at the heart of organizational ethics in health care in this sense: stewardship provides the hermeneutic filter that enables basic ethical principles to be realized practically, within the context of the Catholic theology of work, to concerns in health care. This general argument can shed light on the specific topic of non executive compensation programs as an illustration of organizational ethics in health care. PMID- 11899994 TI - Institutional integrity in Roman Catholic health care institutions. AB - Issues of institutional identity and integrity in Roman Catholic health care institutions have been addressed at the level of individual institutions as well as by organizations of Catholic health care providers and at various levels in the Church hierarchy. The papers by Carol Taylor, C.S.F.N., Thomas Shannon, Kevin O'Rourke, O.P., Gerard Magill in this volume provide a significant contribution to concerns of Roman Catholic health care institutions as they face the challenges of providing health care in a secular, pluralistic, market-driven economy. One way to understand institutional integrity is as a measure of the coherence between what an institution identifies as its commitments (its stated moral character), what an institution does (its manifest moral character) and an institution's fundamental moral commitments (its deep moral character). The essays in this volume support this model of integrity. Although it is not their explicit focus, the four essays together provide a vision of institutional integrity for Catholic health care institutions. Each author focuses on one of the three central aspects of integrity: what one identifies as one's commitments (Taylor), how one's actions reflect one's values (Shannon and Magill), and what one is or what one values at a deep level (O'Rourke). I will offer a brief overview of the ways in which the integrity of Catholic health care institutions has been addressed. Then I will consider the four essays and show how each offers an analysis of one of the three critical elements of integrity. PMID- 11899995 TI - Experimental measurements of generalized grating images. AB - The term generalized grating imaging is used to describe the process of image formation of a grating using only another grating as imaging system. The moire and the Lau effects could be regarded as particular cases of such a process. Here we deal with the less-studied case of images formed at finite distances from the gratings, using an extended monochromatic light source. Some experimental results are shown for the images obtained in this last case, and they are compared with theoretical predictions. PMID- 11899996 TI - Compact red-green-blue beam illuminator and expander. AB - A compact multicolor beam illuminator and a compact multicolor beam expander are presented. The illuminator performs the dual task of demultiplexing a narrow red green-blue (RGB) input beam into three separate beams, each of a different color, and expanding them to yield three separate magnified plane waves. The beam expander expands a narrow RGB input beam into a single magnified RGB plane wave. The design and recording procedures, along with experimental results for a beam illuminator and a beam expander with a magnification of approximately 3, are presented. PMID- 11899997 TI - Compact beam expander with linear gratings. AB - Novel compact beam expanders that could be useful for applications such as providing light to flat panel displays are presented. They are based on a planar configuration in which three spatially linear gratings are recorded on one transparent substrate, so as to expand a narrow incoming beam in two dimensions. We present the design and recording procedures along with results, showing a relatively uniform intensity of the wide output beam. Such expanders can serve for illuminating flat panel displays. PMID- 11899998 TI - Flat-top narrow-band spectral response obtained from cascaded resonant grating reflection filters. AB - Cascaded identical resonant grating reflection filters are shown to exhibit flattened spectral responses when the individual filter elements are cascaded pi out of phase. Off resonance, the net response of a cascaded arrangement is given approximately by the sum of the individual filter responses. Cascading filters pi out of phase thus result in a reduction in the off-resonance reflection levels and correspondingly an increase in the spectral bandpass ratio. The spectral bandpass ratio is a figure of merit used to gauge the flatness of a response and is defined as the ratio of the linewidth at an efficiency of 90% to the linewidth at an efficiency of 10%. Cascading two and three filters in this manner results in respective increases in the spectral bandpass ratio of three times and more than five times that of a single filter. PMID- 11899999 TI - Nematic liquid-crystal polarization gratings by modification of surface alignment. AB - The stylus of an atomic force microscope is used to scribe preferred directions for liquid-crystal alignment on a polyimide-coated substrate. The opposing substrate that comprises the liquid-crystal cell is rubbed unidirectionally, resulting in a twisted nematic structure associated with each micrometer-sized pixel. The polarization of light entering from the uniformly rubbed substrate rotates with the nematic director by a different amount in each pixel, and each of the two emerging polarization eigenmodes interferes separately. Two examples are discussed: a square grating that allows only odd-order diffraction peaks and a grating that combines rotation with optical retardation to simulate a blazed grating for circularly polarized light. The gratings can be electrically switched if used with semitransparent electrodes. PMID- 11900000 TI - Optical properties of end-sealed hollow fibers. AB - We propose sealing techniques for medical hollow fibers to protect the inner surface of fibers from debris or water that scatters from targets. First, hollow fibers are sealed with a film of polymer that is easily formed by use of a dipping technique. The transmission loss of 20-microm-thick sealing film was 0.2 dB for Er:YAG laser light, and the maximum energy that is available for the film was 180 mJ. Second, a sealed glass cap was applied to the output end of hollow fiber. The silica-glass cap with a wall thickness of 400 microm shows a transmission loss of 0.5 dB and was not damaged by radiation of 400-mJ energy pulses. PMID- 11900001 TI - Compact wavelength division multiplexers and demultiplexers. AB - Compact devices for wavelength division multiplexing and demultiplexing, believed to be novel, are presented. These devices are based on planar optics configurations, comprising multiplexed diffractive optical elements. The principle, design, and recording of these planar devices are described, including the fact that the recording is done at a single wavelength in the green region. Experimental procedures and results for planar devices that can handle three wavelengths in the visible as well as in the near infrared are presented. PMID- 11900002 TI - Intrinsic fiber-optic ultrasonic sensor array using multiplexed two-wave mixing interferometry. AB - An intrinsic multiplexed laser interferometer is presented that allows for the simultaneous detection of acoustic waves by an array of fiber-optic sensors. The phase-modulated signals from each sensor are demodulated by use of an adaptive two-wave mixing setup. The light from each sensing fiber in the array is mixed with a reference beam in a single photorefractive crystal (PRC), and the output beams from the PRC are imaged onto separate photodetectors to create a multiplexed two-wave mixing (MTWM) system. The sensing fibers are embedded in graphite-epoxy composite panels, and detection of both acoustic emission and ultrasonic signals in these materials is demonstrated. The intrinsic MTWM system is an effective tool for the simultaneous demodulation of signals from a large fiber sensor array. Also, the adaptive nature of the MTWM setup obviates the need for active stabilization against ambient noise. PMID- 11900003 TI - Ideal concentrators with gaps. AB - Ideal concentrators with large gaps are presented. These new devices use optical elements surrounding the receiver. When their number is large, they (may) constitute a microstructure (many components with small sizes). Smaller gaps can also be achieved by use of a fewer number of optics. Different ways to combine these optical elements are presented for the case of larger and smaller gaps. Designs that use mirrors and total internal reflection are also presented for the case of larger gaps. The mathematical methods used to calculate the shape of the optics are outlined. In spite of the fact that the number of optics may be large, given the symmetries inherent to these designs, the elements in these microstructures are all equal. This is a simplifying feature that is important for their design and eventual production in the future. PMID- 11900004 TI - One-radius triplets. AB - The one-radius triplet is an optical system that consists of three lenses. Either the radii of curvature of the lenses have the same absolute value or one radius of curvature has an infinitely large value. The advantage of such optical systems is that their production cost is less than that of systems with ordinary triplets. Here a theory of one-radius triplets is described and tables of parameters for their modification are provided. Residual aberrations are given for several selected triplets. One-radius triplets are suitable for use in laser technology and metrology. PMID- 11900005 TI - Variation of magnification with focus. AB - Applications of optical systems, including depth by focus and three-dimensional metrology, have been developed recently in which the image is characterized over a range of object depths simultaneously and in which the focus of the systems is also varied. Important to such applications is the variation of image magnification with focus. A general understanding of this phenomenon is developed, and two new and useful concepts associated with the variation of the magnification of an image are introduced. Errors and oversights in the existing literature are explained and corrected, and the requirements for an optical system to exhibit the magnification properties desired in such applications are identified. It is shown that not all telecentric systems have these properties and that there exist practical and attractive nontelecentric systems that do exhibit them. PMID- 11900006 TI - Liquid-crystal micropolarizer array for polarization-difference imaging. AB - Fabrication and applications are discussed for a visible-wavelength micropolarizer array consisting of a linear polarizer and a micropatterned liquid crystal (LC) cell. LC alignment direction is controlled by means of depositing an optically transparent gold film at an oblique angle and coating the surface with an alkanethiol self-assembled monolayer. Microdomains of two perpendicular LC alignment directions are created by photolithography and etching of the gold layer, rotating the substrate 90 deg, and depositing a second oblique gold layer in the etched areas. The resulting array is used for polarization-difference imaging (PDI), a technique that enhances image contrast in the presence of scattering. Images obtained with the array require more processing than do conventional PDI images, but this method eliminates the need for an electronically activated LC filter and is especially suited to systems whose filters are closely integrated with optical sensor arrays. PMID- 11900007 TI - Phasing segmented mirrors: a modification of the Keck narrow-band technique and its application to extremely large telescopes. AB - Future telescopes with diameters greater than 10 m, usually referred to as extremely large telescopes (ELTs), will employ segmented mirrors made up of hundreds or even thousands of segments, with tight constraints on the piston errors between individual segments. The 10-m Keck telescopes are routinely phased with the narrow-band phasing technique. This is a variation of the Shack-Hartmann wave-front sensor in which the signal is the correlation between individual subimages and simulated images. We have investigated the applicability of this technique to ELTs, and in the process we have developed what to our knowledge is a new algorithm in which each subimage provides on its own a piston-dependent value. We also discuss an alternative algorithm to resolve the lambda ambiguity that allows detection of problematic cases, and a modification of the singular value-decomposition procedure used to phase the whole mirror, using weightings on individual measurement errors. By means of simulations we show that the modified technique shows improved performance and that it can work with sufficient precision on telescopes as large as 100 m. PMID- 11900008 TI - Transmission laser microscope using the phase-shifting technique and its application to measurement of optical waveguides. AB - A type of a transmission phase-shifting laser microscope, believed to be new, has been developed. In this microscope a biprism located between a magnifying lens and an observation plane was used as a beam splitter. The biprism is laterally translated to introduce phase shifts required for quantitative phase measurement with a phase-shifting technique. The disturbance caused by a Fresnel-diffracted wave from the splitting edge of the biprism is reduced by placement of a linear beam stopper at the center of an intermediate image plane. As the first application, the developed microscope is used to measure a refractive-index distribution in optical waveguides. A difference of refractive indices of less than 6 x 10(-5) is clearly measured in the submicrometer region. PMID- 11900009 TI - Low-coherence interferometer system for the simultaneous measurement of refractive index and thickness. AB - We have developed a low-coherence interferometer system used for the simultaneous measurement of refractive index n and thickness t of transparent plates. Both the phase index n(p) and group index n(g) can be determined automatically in a wide thickness range of from 10 microm to a few millimeters. Two unique techniques are presented to measure n(p), n(g), and t simultaneously. One allows us to determine n(p), n(g), and t accurately by using a special sample holder, in which the measurement accuracy is 0.3% for the thickness t above 0.1 mm. In the other technique the chromatic dispersion delta n of index is approximately expressed as a function of (n(p) - 1) on the basis of measured values of n(p) and n(g) for a variety of materials, and then the simultaneous measurement is performed with a normal sample holder. In addition, a measurement accuracy of less than 1% is achieved even when the sample is as thin as 20 microm. The measurement time is also 3 min or more. PMID- 11900010 TI - Optical in-plane strain field sensor. AB - A whole-field speckle strain sensor is presented. The speckle strain sensor allows the measurement of all three in-plane components of the strain field simultaneously without touching the surface of the sample. The strain fields are extracted from the in-plane motion of defocused laser speckles in a telecentric imaging system. To distinguish the contribution to the speckle motion from surface translation, rotation, and strain, the speckle motion from three lasers with different illumination directions and wavelengths has to be analyzed separately. Simultaneous acquisition of the three individual speckle patterns is achieved by means of splitting the light from the lasers onto separate but synchronized detectors with the aid of dichroic mirrors. The motion of the speckles is calculated with digital speckle photography (speckle correlation), which enables the strain sensor to measure strain fields with noise levels as low as 10 microstrain. PMID- 11900011 TI - Autocompensating interferometer for measuring the changes in refractive index of supercooled water as a function of temperature at 632.8 nm. AB - An interferometric arrangement that automatically compensates for thermal expansion was used to examine the changes in the refractive index of liquid water at 632.8 nm as a function of temperature from 10 to -15 degrees C. By combining the results of this research with existing data, we calculated the absolute refractive index in the supercooled region with an accuracy ranging from 3 x 10( 6) to 1 x 10(-6). A direct observation of the refractive-index maximum of water is reported for the first time to our knowledge and found to occur between 0 and 0.1 degrees C. PMID- 11900012 TI - Development of an interferometer for measurement of the diffusion coefficient of miscible liquids. AB - A common-path interferometer (CPI) system was developed to measure the diffusivity of transparent liquid pairs by real-time visualization of the concentration gradient profile. The CPI is an optical technique that can be used to measure changes in the gradient of the refractive index of transparent materials. The CPI is a shearing interferometer that shares the same optical path from a laser light source to the final imaging plane. Molecular diffusivity of liquids can be determined by use of physical relations between changes in the optical path length and the liquid phase properties. The data obtained by this interferometer are compared with similar results from other techniques. This demonstrates that the instrument is reliable for measurement of the diffusivity of miscible liquids and allows the system to be compact and robust. It can also be useful for studies in interface dynamics as well as other applications in a low-gravity environment. PMID- 11900013 TI - SHIMMER: a spatial heterodyne spectrometer for remote sensing of earth's middle atmosphere. AB - It is well known and demonstrated that interference spectroscopy offers capabilities to obtain passive remote optical sensing spectra of high precision and also achieves economies in size, cost, and ease of deployment compared with more conventional systems. We describe the development of a near-ultraviolet spatial heterodyne spectrometer designed for remote sensing of the global distribution of the hydroxyl radical OH in the Earth's middle atmosphere. The instrument, known as SHIMMER (Spatial Heterodyne Imager for Mesospheric Radicals), is expected to obtain its first OH measurement from space in early 2002 from the Space Shuttle. PMID- 11900014 TI - Experimental comparison of a liquid-crystal point-diffraction interferometer (LCPDI) and a commercial phase-shifting interferometer and methods to improve LCPDI accuracy. AB - We compare the phase measurements of a fused-silica witness sample made with a liquid-crystal point-diffraction interferometer (LCPDI) with measurements made with a Zygo Mark IV xp phase-shifting interferometer and find close agreement. Two phase-shift-error sources in the LCPDI that contribute to measurement discrepancies are frame-to-frame intensity changes caused by the dichroism of the dye and alignment distortions of the host liquid crystal. An empirical model of the phase-shift error caused by the host alignment distortions is presented and used to investigate the performance of two different phase-detection algorithms. It is suggested that by proper choice of LCPDI fabrication parameters and phase acquisition methods, the device's accuracy can be significantly improved. PMID- 11900015 TI - Dispersion of barium gallogermanate glass. AB - Gallogermanate glasses are the subject of intense study as a result of their unique combination of physical and optical properties, including transmission from 0.4 to beyond 5.0 microm. These glasses can be easily made into large optics with high-index homogeneity for numerous U.S. Department of Defense and commercial visible-IR window applications such as reconnaissance, missile domes, IR countermeasures, avionics, and collision avoidance on automobiles. These applications require a knowledge of the refractive index of glass throughout the region of transmission. Consequently, we have measured the refractive index of BaO-Ga2O3-GeO2 glass from 0.4 to 5.0 microm and calculated the Sellmeier coefficients required for optical device design. PMID- 11900016 TI - Digital speckle photography: visualization of mesoflow through clustered fiber networks. AB - Digital speckle photography (DSP) is used for velocity field measurements inside a fiber network. The width of the channels in which the flow is measured is typically less than 1 mm. Therefore a microscope is used to image the fiber network. When we sample 30 images/s and separate the moving parts of the images from the stationary parts, the velocity field can be deduced with DSP. PMID- 11900017 TI - Risley prisms to control wave-front tilt and displacement in a vectorial shearing interferometer. AB - A pair of thin prisms is used to deviate a light beam without changing the image orientation in a vectorial shearing interferometer. The relative angle between prisms determines the displacement of the wave front and its tilt. The direction of the beam displacement is controlled by means of changing the relative angle between prisms. This system is employed to control the displacement of a sheared wave front as a vector quantity and to introduce a controlled amount of tilt in what we believe is a novel interferometric shearing system. The predicted performance of this wave-front director is confirmed experimentally. PMID- 11900018 TI - Multilayer ARROW channel waveguide for evanescent field enhancement in low-index media. AB - A multilayer antiresonance reflecting optical waveguide (ARROW) channel waveguide geometry, believed to be novel, is proposed for enhancing the evanescent field in low-index materials. The finite-difference method is used in the analysis of the structure. The fraction of the fundamental TE-like-mode power in the low-index material (air) is used as a measure of the evanescent field enhancement. The calculated results suggest that the evanescent field of the fundamental TE-like mode can be significantly increased in air while the low modal loss that characterizes the leaky nature of the structure is maintained. The results also suggest that a semivectorial approach to this problem is adequate for analysis of the proposed waveguide structure. PMID- 11900019 TI - Theoretical considerations for arrayed waveguide displays. AB - An arrayed waveguide display (AWD) is proposed in which a linear emitter array is coupled to a waveguide array upon which liquid-crystal (LC) switches are formed. Images are displayed by line-by-line control of light, including emission by the emitter array, injection into the waveguide array, propagation inside, and extraction by use of the LC switches. With mostly polymeric materials, an AWD can be made thin and tough. Its light-use efficiency can be made larger than that of a conventional transmissive LC display. Because of these potential advantages, an AWD is well suited for mobile applications. PMID- 11900020 TI - Compact static Fourier transform spectrometer with a large field of view based on liquid-crystal technology. AB - We present designs of static Fourier transform spectrometers that are based on a Wollaston prism with a large field of view. Besides the usual advantages of static Fourier spectrometers (large resolving power, large wave-number range, high throughput), these designs also present the advantage of using relatively cheap liquid-crystal technology. The use of twisted liquid-crystal structures gives a large field of view, which in turn gives the ability to collect more light from a divergent light source. Measurements are compared with simulations. Different simulation principles are used. We found new configurations by using twisted structures that show a large field of view. PMID- 11900021 TI - Light scattering by aggregated red blood cells. AB - In low flow rates, red blood cells (RBCs) fasten together along their axis of symmetry and form a so-called rouleaux. The scattering of He-Ne laser light by a rouleau consisting of n (2 < or = n < or = 8) average-sized RBCs is investigated. The interaction problem is treated numerically by means of an advanced axisymmetric boundary element--fast Fourier transform methodology. The scattering problem of one RBC was solved first, and the results showed that the influence of the RBC's membrane on the scattering patterns is negligible. Thus the rouleau is modeled as an axisymmetric, homogeneous, low-contrast dielectric cylinder, on the surface of which appears, owing to aggregated RBCs, a periodic roughness along the direction of symmetry. The direction of the incident laser light is considered to be perpendicular to the scatterer's axis of symmetry. The differential scattering cross sections in both perpendicular and parallel scattering planes and for all the scattering angles are calculated and presented in detail. PMID- 11900022 TI - Elastic light scattering with a LiNbO3 waveguide. AB - We present the measurements of hemispherical elastic scattering of light guided by different modes of a planar LiNbO3 waveguide. It is shown that the fundamental and the lowest-order modes are more sensitive to the scattering properties of the air-core interface, whereas higher-order modes are more sensitive to the optical inhomogeneities of the core-substrate interface. We also demonstrate that because of static polarization of LiNbO3 crystal, the air-core interface is sensitive to the presence of dust particles in air, which causes a change in the scattered light based on time of observation. This sensitivity could be used to elaborate compact sensors of air contamination. PMID- 11900023 TI - Radiometry in line-shape modeling of Fourier-transform spectrometers. AB - A radiometric model of the instrument line shape (ILS) of Fourier-transform spectrometers is presented. We show first that common line-shape models are based on distribution of the radiant intensity in the interferometer. The complete steps between the source and the ILS are exposed as the core of the model. Relationships between the ILS, the spectrum as measured by the instrument, and the spectrum as emitted by the scene are demonstrated from the ILS model. Then the formal radiometric modeling of the ILS is derived, including the contribution of the aperture of the optical system. The particular case of a centered circular aperture with a uniform Lambertian radiance in the field of view is discussed. Conditions are deduced to ensure that the only spectral variation of the ILS is a scaling with wave number, as is usually assumed in current line-shape models. The ILS dependence on the scene is also discussed, and the effect of taking into account the radiometry on the ILS is estimated for the case of an ideal thin lens used as a collimator. PMID- 11900024 TI - Efficiencies of master, replica, and multilayer gratings for the soft-x-ray extreme-ultraviolet range: modeling based on the modified integral method and comparisons with measurements. AB - The near-normal-incidence efficiencies of a 2400-groove/mm holographic master grating, a replica grating, and a multilayer grating are modeled in the soft-x ray-extreme-ultraviolet (EUV) regions and are compared with efficiencies that are measured with synchrotron radiation. The efficiencies are calculated by the computer program PCGrate, which is based on a rigorous modified integral method. The theory of our integral method is described both for monolayer and multilayer gratings designated for the soft-x-ray-EUV-wavelength range. The calculations account for the groove profile as determined from atomic force microscopy with a depth scaling in the case of the multilayer grating and an average random microroughness (0.7 nm) for the short wavelengths. The refractive indices of the grating substrate and coatings have been taken from different sources because of the wide range of the wavelengths (4.5-50 nm). The measured peak absolute efficiency of 10.4% in the second diffraction order at a wavelength of 11.4 nm is achieved for the multilayer grating and is in good agreement with a computed value of approximately 11.5%. Rigorous modeling of the efficiencies of three similar gratings is in good overall agreement with the measured efficiency over a wide wavelength region. Additional calculations have indicated that relatively high normal incidence efficiency (of at least several percent) and large angular dispersion in the higher orders can be achieved in the 4.5-10.5-nm range by application of various multilayer coatings. PMID- 11900025 TI - Optical design of the wide angle camera for the Rosetta mission. AB - The final optical design of the Wide Angle Camera for the Rosetta mission to the P/Wirtanen comet is described. This camera is an F/5.6 telescope with a rather large 12 degrees x 12 degrees field of view. To satisfy the scientific requirements for spatial resolution, contrast capability, and spectral coverage, a two-mirror, off-axis, and unobstructed optical design, believed to be novel, has been adopted. This configuration has been simulated with a ray-tracing code, showing that theoretically more than 80% of the collimated beam energy falls within a single pixel (20" x 20") over the whole camera field of view and that the possible contrast ratio is smaller than 1/1000. Moreover, this novel optical design is rather simple from a mechanical point of view and is compact and relatively easy to align. All these characteristics make this type of camera rather flexible and also suitable for other space missions with similar performance requirements. PMID- 11900026 TI - Method for reducing background artifacts from images in single-photon emission computed tomography with a uniformly redundant array coded aperture. AB - Uniformly redundant array coded apertures have proven to be useful in the design of collimators for x-ray astronomy. They were initially expected to be equally successful in single-photon emission computed tomography (SPECT). Unfortunately, the SPECT images produced by this collimator contain artifacts, which mask the true picture and can lead to false diagnosis. Monte Carlo simulation has shown that the formation of a composite image will significantly reduce these artifacts. A simulation of a tumor in a compressed breast phantom has produced a composite image, which clearly indicates the presence of a 5 mm x 5 mm x 5 mm tumor with a 6:1 intensity ratio relative to the background tissue. PMID- 11900027 TI - Feasibility study of a system for combined light dosimetry and interstitial photodynamic treatment of massive tumors. AB - A system for the photodynamic laser treatment of massive tumors that employs multiple optical fibers to be inserted into the tumor mass is described. The light flux through the tumor can be assessed by use of the individual fibers both as transmitters and as receivers. With a computer model that describes the diffusive light propagation, optical dosimetry is under development. The system has been tested in an experimental animal tumor model in preparation for clinical work. Currently, delta-aminolevulinic acid is used as a sensitizer, activated by 635-nm radiation from a 2.0-W compact diode laser system. With the availability of future, highly selective drugs absorbing approximately 750 nm, larger tumor volumes should be treatable, and surrounding, sensitive normal tissue should be spared. PMID- 11900029 TI - Pulse oximetry: its invention, contribution to medicine, and future tasks. PMID- 11900028 TI - Determination of glucose concentration in a scattering medium based on selected wavelengths by use of an overtone absorption band. AB - A method and device for measuring glucose concentration in a scattering medium have been developed. A spectral range of 800-1800 nm is considered for wavelength selection because of its deeper penetration into biological tissue and the presence of a glucose absorption band. An algorithm based on selected wavelengths is proposed to minimize interference from other components. The optimal distance between the light source and the detector for diffuse reflectance measurement minimizes the influence of medium scattering. The proposed algorithm and measuring device are tested with a solution containing milk with added glucose. Glucose concentrations between 0 and 2000 mg/dl are determined with a correlation coefficient of 0.977. We also investigate the influence of concentration variations of other substances such as water, hemoglobin, albumin, and cholesterol when they are mixed in a scattering medium. PMID- 11900031 TI - Standardization of the testing of pulse oximeter performance. PMID- 11900030 TI - Accuracy of pulse oximeters: the European multi-center trial. PMID- 11900032 TI - Design and validation of pulse oximetry for low saturation. AB - As pulse oximetry migrates into new areas of patient care, it becomes increasingly important to rigorously test performance in the environment of use before claiming success. If evaluated only under tightly or ideally controlled laboratory or bench-top conditions, results may have little relationship to real world behavior. As an example, fetal pulse oximetry sensors are subject to challenges not typically found with air-breathing patients. The "presenting part" is an attractive site for fetal oximetry, however the presence of resolvable pulses by itself does not ensure accurate readings. This study explores the effects of physiological and mechanical perturbations at low saturation expected during use on fetuses with such sensors. Numerical modeling and animal studies demonstrate that at 40% true arterial saturation (SaO2), calculated saturation values (SpO2) may err by more than 30%, even in the presence of otherwise normal pulse oximetry signals. PMID- 11900033 TI - Multi-wavelength reflectance pulse oximetry. AB - The performance of current reflectance pulse oximeters is hindered by poor signal to-noise ratio. To overcome this problem a new reflectance oximeter has been developed with a sensor which consists of three LEDs and two continuous photodetector rings placed equidistant from the center of the LEDs. In addition, ultra low noise electronics and adaptive algorithm assure improved performance. A validation study was performed on 10 healthy volunteers. Sensors were placed on several sites and measurements were compared to reference arterial blood samples. During the study progressive hypoxemia was induced by lowering the inspired oxygen concentration (FiO2) to 10%, followed by a recovery phase. Twelve blood samples were taken during each cycle, yielding a total of 120 measured data points. Data from randomly selected 5 subjects was used for calibration and subsequently tested on the other 5 subjects. Results proved to be well within clinically acceptable boundaries for all 3 sampling sites with high correlation (R2 > 0.9) and SD around 2%. In conclusion, a new 3 wavelength reflectance pulse oximeter with unique sensor geometry and improved algorithms provides enhanced performance and is less susceptible to poor signal to noise conditions when compared to existing reflectance oximetry systems. PMID- 11900034 TI - Assessing high-risk infants in the delivery room with pulse oximetry. AB - Heart rate and oxygenation status are essential to the delivery room assessment of newborn infants. Pulse oximetry (PO) can capture these two variables but low peripheral perfusion, patient motion and the presence of ambient lighting can challenge the technology. New generation pulse oximeters claim to measure through these conditions. The objective of this study was to assess the performance of new generation pulse oximeters during the delivery room assessment and management of high-risk newborns. Part one (study A) compared the outcome of 25 infants simultaneously monitored with two new PO technologies (experimental group) to a matched population of 25 infants without PO-monitoring (control group). Findings from the experimental group revealed a significant difference between new PO technologies and an improvement in patient outcome compared to the control group. Part two (study B) evaluated the affects of PO-monitoring on 15 very immature infants. Use of Masimo SET PO was associated with a rapid acquisition and near continuous display of PO values, which proved valuable for initial respiratory care and to determine the need for more intensive procedures. Both studies revealed that near immediate postnatal PO monitoring of newborns in the delivery room was feasible and valuable. PMID- 11900035 TI - Use of pulse oximetry in automated oxygen delivery to ventilated infants. PMID- 11900036 TI - Designing a pulse oximeter safety standard. AB - The development of a safety standard is a time-consuming and complex task. Many factors influence the stringency of the requirements, including history with the products and test methods, safety records, and the intended use of the standard and device. Four primary issues are used to illustrate the complexity of developing a safety standard for today's complicated software-controlled pulse oximeters: Can a patient simulator be used to assess performance? What is meant by motion artifact resistance? What is a safe surface temperature limit for the probe? What default low SpO2 limit should be required? Under ASTM's new standards development paradigm, ASTM may achieve consensus on the 2001 draft of ASTM F1415 but not publish it, electing instead to submit the standard to ISO for consideration as the internationally harmonized pulse oximeter safety standard. PMID- 11900037 TI - Pulse oximetry in the neonatal intensive care unit (NICU): detection of hyperoxemia and false alarm rates. PMID- 11900038 TI - Pulse oximetry in the management of children in the PICU. AB - The noninvasive and continuous nature of pulse oximetry is a considerable asset inpatient monitoring in the PICU. Its use in the PICU is constantly challenged by the combined presence of vigorous body motion and poor peripheral circulation, conditions which demand special consideration in developing a better pulse oximeter. This is a point of fierce competition among manufacturers. PMID- 11900039 TI - Differences in pulse oximetry technology can affect detection of sleep-disorderd breathing in children. AB - Newer pulse oximeters have been developed to be motion resistant and thus have few false alarms. However, they have not yet been evaluated in a pediatric sleep laboratory setting. While evaluating new oximeters for use in our laboratory, we obtained simultaneous pulse oximetry data from two Masimo oximeters and from two Nellcor oximeters during nocturnal polysomnography in children referred for sleep disordered breathing (SDB). In series 1, comprising 24 patients, comparisons were made between a Masimo oximeter with 4-second averaging time and the Nellcor N-200 oximeter set for 3 to 5 second averaging. A maximum of 20 events per patient were randomly selected for analysis, an "event" being a desaturation of > or = 4% registered by either oximeter. Interobserver agreement for event classification was 93%. Eighty-eight percent of 220 desaturation events occurring during wakefulness and 38% of 194 events occurring during sleep were classified as motion artifact on the Nellcor oximeter. Neither the Masimo oximeter nor the transcutaneous oxygen probe confirmed that the desaturation was real, in most of these cases. During sleep, there were 119 events detected by either or both oximeters: 113 (95%) by the Nellcor versus 82 (69%) by the Masimo. For these 119 events, the extent of desaturation was slightly less for the Masimo than the Nellcor oximeter, 4.5 +/- 2.4% versus 5.5 +/- 2.5%, respectively. In series 2, 22 patients were studied comparing a Masimo Radical oximeter with 2 second averaging to the Nellcor N-200 oximeter. The extent of desaturation was slightly greater for the Masimo oximeter. The Masimo oximeter detected more non-artifactual desaturation events occurring during sleep than the Nellcor oximeter, 90% versus 76% (chi2 = 9.9, p < 0.01). In series 3, comprising 128 events in 5 patients, a Nellcor N-395 oximeter detected fewer desaturations during non-movement, sleep periods and had more movement related "desaturation" events, compared to a Masimo Radical oximeter. CONCLUSIONS: The Masimo oximeters register many fewer false desaturations due to motion artifact. Using 4-second averaging, a Masimo oximeter detected significantly fewer SaO2 dips than the Nellcor N-200 oximeter but using 2-second averaging, the Masimo oximeter detected more SaO2 dips than the Nellcor N-200 oximeter. The sensitivity and motion artifact rejection characteristics of the Nellcor N-395 oximeter are not adequate for a pediatric sleep laboratory setting. These findings suggest that in a pediatric sleep laboratory, use of a Masimo oximeter with very short averaging time could significantly reduce workload and improve reliability of desaturation detection. PMID- 11900040 TI - A characterization of motion affecting pulse oximetry in 350 patients. AB - As part of an oximetry research effort in which plethysmographic data were collected from moving patients, a wide variety of patient motion affecting pulse oximetry was observed and characterized by the clinical incidence, type, severity and duration of patient motion. 350 patients were observed for movement in clinical settings, including ICU, SICU, MICU, PICU, NICU, OR and PACU, at 4 hospitals and on an ambulance. 20% (70) of the patients exhibited motion. Half (35) of the moving patients were instrumented to record oximetric, plethysmographic and/or acceleration information. 31% of NICU infants moved compared to only 7% of adults in ICUs. The most common noisy oximetry signals were caused by motion characterized by extending/flexing (/kicking in infants) and by clenching/pressing/rubbing. In infants, these motion types accounted for 53% and 11% of motion, respectively. Less common infant motion types were patient cares, shifting body position and cough/cry. The most common adult motions were equally divided among extend/flex, clench/press/rub, twitch/shake and transport motion types. Extend/flex motions typically demonstrated high plethysmographic waveform modulation (71.5% maximum) and high acceleration. Clench/press/rub motions typically also had high modulations, but low G-force. With one high-G exception, twitch/shake motions had little or no effect on oximetry readings. Less common adult motion types affecting pulse oximetry included cough/cry, strain/posture, tremors and tap/bump. Most recorded motions were aperiodic and short-lived, 62% lasting less than 10 seconds, only 5% lasting over 1 minute. NICU patients made the longest lasting continuous series of motions, while adults made 86% of the motions lasting a second or less. PMID- 11900041 TI - Issues in the laboratory evaluation of pulse oximeter performance. PMID- 11900042 TI - False alarm rates of three third-generation pulse oximeters in PACU, ICU and IABP patients. AB - The objective of this clinical study was to determine alarm rates--in particular the frequency of false positive alarms--of three third-generation pulse oximeters in the postanesthesia care unit (PACU), the intensive care unit (ICU), and in patients with an intra-aortic balloon pump (IABP): Nellcor Symphony N-3000, a Masimo IVY 2000, and Agilent Viridia CMS 2000. All alarms were classified into technical/physiological and false/correct. 235 consecutive ASA physical status I IV patients after surgery were included into the study. In the PACU false positive alarms were rare: CMS n = 60, N-3000 n = 60, Masimo n = 87. Bland-Altman testing discovered only negligible differences of alarm rates and dropout times. Out of a total of 728 alarms 67.3% were classified as false positive in ICU patients: 97 alarms by CMS, 176 by N-3000 and 218 by Masimo SET. If IABP was present, CMS indicated a significant smaller number of false positive alarms (n = 35, 7.2%) when compared to Masimo SET (n = 188, 38.9%) and N-3000 (n = 229, 47.4%), consecutively the majority of false positive alarms (76.2%) can be rated as a result of the interference of IABP. Unless IABP (and to a considerably smaller extent cardiac arrhythmia) is present the pulse oximeters do not differ significantly regarding sensitivity and specificity. PMID- 11900043 TI - Combining transcutaneous blood gas measurement and pulse oximetry. AB - We are describing the preliminary results of tests performed in adult volunteers and in adult patients during and after general anesthesia with a miniaturized single sensor combining the continuous and non-invasive measurement of oxygen saturaiton by pulse oximetry (SpO2) and transcutaneous PCO2 (OxiCarbo sensor). The sensor is heated to 42 degrees C to arterialize the cutaneous tissue and is applied at the ear lobe with a special low-pressure clip. The results indicate a good agreement between ear lobe PCO2 and arterial PCO2 in the range 35 to 70 mmHg (10 patients, number of measurements 104, regression line TcPCO2 = 1.01 PaCO2 + 0.59 mmHg, bias 1.22 mmHg, SD 3.69 mmHg) and between ear lobe SpO2 and SaO2 (bias 0.44% with SD 0.77% in the range 80% to 100%, bias 1.39% with SD 1.43% in the range 60% to 80%). The ear lobe OxiCarbog sensor detects the SpO2 change 5 to 37 sec faster than a finger sensor and the PCO2 change 9 to 48 sec faster than a transcutaneous sensor fixed at the upper arm. Further improvements versus single sensors are a higher stability of the SpO2 signal and the possibility of performing long term SpO2 and PCO2 measurement at the ear lobe. PMID- 11900045 TI - Advantages of new technology pulse oximetry with adults in extremis. AB - Thirteen critically ill patients in the thoracic and cardiovascular ICU in whom conventional pulse oximetry (CPO) failed were identified and prospectively studied using an oximeter employing Masimo SET technology (MSO). These patients were elderly and ill, having undergone a variety of thoracic and cardiovascular operations. They were often hypothermic and shivering. Several had arterial assist devices, intraaortic balloon pumps or impending or actual cardiac arrest. In 12 of these patients the MSO was able to determine the actual arterial saturation as calculated from a coincident arterial blood gas. In the one patient, in whom no saturation was obtained with the MSO, no arterial blood could be obtained due to complete cardiac collapse. This case series demonstrates the value of the new Masimo SET technology for pulse oximetry in the most critically ill and demanding clinical situations in which other techniques have failed. A significant improvement in patient safety resulted from use of this improved oximetry. PMID- 11900044 TI - Design and validation of a pulse oximeter calibrator. AB - The performance of a new calibrator for pulse oximeters is tested with five pulse oximeters from different manufacturers. The calibrator is based on time resolved transmission spectra of human fingers. Finger spectra with different arterial oxygen saturation can be selected to simulate real patients. The results obtained with this calibration device are compared with the results of conventional calibration procedures with volunteers. Beside accuracy tests the suitability for artifact simulation with the new device is discussed. The response of the five tested pulse oximeters is in good agreement with the response of the pulse oximeters connected to real patients. A test procedure for pulse oximeters similar to the conventional desaturation practice is possible; some of the typical artifacts pulse oximetry has to cope with can be simulated easily. PMID- 11900046 TI - Interpretive oximetry: future directions for diagnostic applications of the SpO2 time-series. PMID- 11900048 TI - The theory and applications of pulse spectrophotometry. AB - Opening of the first door of pulse photometry gave us pulse oximeter. The next door opens to multiwavelength pulse photometry. It will give us a high performance pulse oximeter, providing a wide variety of clinical information simultaneously. This next generation of pulse photometry should further improve bedside monitoring and patient care. PMID- 11900047 TI - New developments in the measurement of CO-oximetry. AB - The CO-oximeter (HEMOXIMETER) in the ABL 700 is described with respect to construction and parameters. Factors affecting stability and accuracy are discussed. Artificial blood substitutes and their effect on the oximetric data are discussed. PMID- 11900049 TI - When pulse oximetry monitoring of the critically ill is not enough. PMID- 11900050 TI - Histoplasma capsulatum var. capsulatum occurring in an HIV-positive Ghanaian immigrant to Italy. Identification of H. capsulatum DNA by PCR from paraffin sample. AB - Histoplasmosis, which is highly endemic in the United States, is rare in Europe, usually imported but sometimes autochthonous. In Africa, histoplasmosis capsulati coexists with "African histoplasmosis", a characteristic skin infection caused by H. capsulatum var. duboisii. Histoplamosis due to H. capsulatum is one of the 12 secondary infections listed in the surveillance definitions of AIDS. We report the case of a 36-year-old black man with acquired immunodeficiency syndrome (AIDS) who was living in Italy but originally came from Ghana. Histoplasmosis was disseminated with fever and cutaneous manifestations. The diagnosis was demonstrated morphologically based on the presence of yeast, observed by light microscopy, in skin lesions and by identification of H. capsulatum var. capsulatum DNA by nested PCR from a paraffin sample. No clinical reports of histoplamosis capsulati in Ghana have been published until now. The present case stresses the role of immigration of subjects from outside Europe who have been infected in their native country. PMID- 11900051 TI - Proliferation conditions for human satellite cells. The fractional content of satellite cells. AB - Primary satellite cell cultures have become an important tool as a model system for skeletal muscles. A common problem in human satellite cell culturing is fibroblast overgrowth. We combined N-CAM (Leu19) immunocytochemical staining of satellite cells (Sc) with stereological methods to estimate the fraction of Sc in culture. Evaluation of different culture conditions allowed us to find proliferation conditions preferentially for Sc: a) Sc should be cultured on surfaces coated with ECM-gel. b) Primary cell culture should be inoculated in DMEM supplemented with 10% fetal calf serum to increase cell adherence. c) Change of media to DMEM supplemented with 2% Ultroser-G and 2% FCS after 24 h.d) Before subcultivation, cells should be preplated for 30 min. The fractional content of Sc in passage four when applying this method of cultivation was 0.82 +/- 0.07 (mean +/- SE, N = 10). Our method enabled us to establish culture conditions which resulted in high Sc content despite several subcultivations. Estimation of the fractional cell content could be a useful tool for optimizing not only Sc culturing but all cultures initially containing more cell types. PMID- 11900052 TI - A cellular model system of differentiated human myotubes. AB - The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc-cultures. In order to identify the differentiation conditions which give a good survival of myotubes and a high grade of differentiation, Sc-cultures were induced to differentiate in media supplemented with either 2% fetal calf serum (FCS) 2% horse serum (HS) or 10% HS. Based on higher CK-activities in cultures differentiating in FCS supplemented media compared to horse sera, fetal calf serum was chosen to induce differentiation. The ATP, DNA and protein content increased during the first 4 days after induction of differentiation and was followed by a period with minor changes. The maximal differences of ATP, DNA and protein between days 4-10 were evaluated and the differences in the three components were found to be less than 20% of the average value with a certainity of more than 0.9. Day 8-myotubes were investigated morphologically and were found immunoreactive for fast myosin, and expressed areas with clear cross striation. We recommend the use of differentiated Sc-cultures in the period from day 4 to 8 after induction of differentiation as only minor differentation-related changes will take place in the cells during this period of time. PMID- 11900053 TI - High prevalence rates of adult silent coeliac disease, as seen in Sweden, must be expected in Denmark. AB - AIM: To disclose the prevalence of adult "silent" coeliac disease in Denmark and Sweden. EXPERIMENTAL DESIGN: 1573 Danish and 1866 Swedish healthy blood donors were screened for the presence of serum anti-gliadin antibodies (AGA) by enzyme linked immunosorbent assay. AGA-positive serum samples were further analysed for IgA anti-endomysium antibodies (EmA) by indirect immunofluorescence microscopy. MAIN RESULTS: The Danish donor population had a higher mean age than the Swedish (41.4 years versus 37.6 years) and a higher proportion of females (41% versus 32%), and had a lower mean level of AGA (17.3 units versus 20.6 units). Sixty-one (3.9%) Danish donors had AGA above the cut-off limit, and four of these also had positive EmA tests. Sixty (3.2%) Swedish donors had AGA above the cut-off limit, and five of these also had positive EmA. Coeliac pathology was proven by biopsy in all five coeliac disease-suspected Swedish donors. No small intestinal biopsy was performed in the coeliac disease-suspected Danish donors. CONCLUSIONS: Based upon the finding of EmA in AGA-positive serum samples, silent coeliac disease may be suspected in 1 per 394 Danish blood donors (2.5 per 1,000). A similar rate was proven in 1 per 373 Swedish blood donors (2.7 per 1,000), indicating no major differences in the prevalence of adult silent coeliac disease between the two neighbouring countries. PMID- 11900055 TI - Stereological study of the cells of dorsal root ganglia in male diabetic rats. AB - Most research on diabetes mellitus has focused on physiological and biochemical aspects of the peripheral nervous system, whilst little work has been done on morphological changes of the neurons. In the present study the effects of diabetes mellitus on cervical and lumbar dorsal root ganglia (C7 and L5) were investigated using modern stereological methods. Twelve adult male Sprague-Dawley rats were randomly divided into two groups. Each group contained six male rats. Diabetes was induced in the experimental group by intraperitoneal injection of 50 mg/kg streptozotocin. At the end of 6 weeks, the rats were fixed by whole body perfusion transcardially with a buffered formalin solution. The seventh cervical and fifth lumbar dorsal root ganglia were removed and immersed in buffered formalin. After tissue processing, the ganglia were embedded in cylindrical paraffin blocks. Isotropic uniform random sections were obtained using the orientator method. Sections (5 microm thick) were selected and stained with Heidenhain's azan. Volume of perikarya of A- and B-cells and their nuclei was estimated using the nucleator method. Before estimating the mean volume, the cells were sampled using the physical disector and point sampling method. Measurements showed that mean perikaryal and nuclear volume of A- and B-cells of dorsal root ganglia (C7 and L5) was reduced in diabetic rats (p<0.05). B-cell mean perikaryal volume in diabetic rats and A- and B-cell mean nuclear volume were reduced by 66% on average. The mean volume of A-cell perikarya was affected less than the others (average 33%). In addition, the difference between the perikaryal and nuclear volume of the seventh cervical and fifth lumbar dorsal root ganglia was not statistically significant. The present study, using stereological techniques, demonstrates reduced perikaryal and nuclear volume of the seventh cervical and fifth lumbar dorsal root ganglia in diabetic rats. PMID- 11900054 TI - Renal structures in type 2 diabetic patients with elevated albumin excretion rate. AB - Renal biopsies were obtained from type 2 diabetic patients with elevated albumin excretion. The aim was to obtain quantitative structural data to correlate with clinical findings. Biopsies from 27 diabetic patients and 12 non-diabetic cases were analysed. Stereological methods were applied by light- and electron microscopy. Diabetic patients showed quantitatively markedly expressed diabetic glomerulopathy, but also an increase in glomerular volume, in prevalence of new vessel formation at the vascular pole, prevalence of glomerular occlusion and in interstitial volume fraction. A significant correlation was not observed between the degree of interstitial and glomerular involvement. The glomerular hypertrophy is interpreted as a compensatory phenomenon, leading to preservation of filtration surface in the open glomeruli. Close correlation was seen between glomerulopathy and glomerular function, and also with the stage of retinopathy. New vessel formation at the vascular pole was most frequent in patients with proliferative retinopathy. Signs of non-diabetic glomerulopathy were not observed, but various atypical ultrastructural changes accompanying the advanced stages are illustrated. Our present findings correspond to data from type I diabetic patients. It is emphasised that all compartments of the kidney are affected by the diabetic state. It is suggested that the interstitial and glomerular lesions are influenced by different factors. PMID- 11900057 TI - Malignant neoplasms: discordance between clinical diagnoses and autopsy findings in 3,118 cases. AB - BACKGROUND: During the past few decades, hospital autopsy rates have steadily declined throughout the Western world. This decline is mainly attributed to the introduction of advanced diagnostic techniques. Despite technological developments, discrepancy rates between clinical diagnoses and autopsy findings remain high. Few studies have addressed discrepancy rates exclusively with regard to malignant neoplasms. In the present study, we reviewed the records of 3,118 autopsies performed at Mayo Clinic during a 6-year period (1994-1999) and identified clinically undiagnosed malignancies found at autopsy and clinically diagnosed cancers not confirmed at postmortem examination. MATERIALS AND METHODS: Autopsy protocols, provisional and final anatomic diagnoses, and data from the Mayo Autopsy Pathology Quality Assurance program were reviewed in an attempt to identify discrepancies between clinical diagnoses and autopsy findings regarding malignant neoplasms. RESULTS: In 3,118 autopsies performed at Mayo Clinic between 1994 and 1999, a malignant tumor was identified in 768 cases (25%). In 128 of 3,118 cases (4.1%), the malignancy was not diagnosed clinically. In 14 of 3,118 cases (0.45%), autopsy failed to confirm a clinically diagnosed cancer. A review of the literature is presented. CONCLUSIONS: Autopsy remains an effective tool for the confirmation and refutation of clinical diagnostic findings regarding malignant neoplasms. PMID- 11900056 TI - Long-term studies of the juxtaglomerular apparatus in young microalbuminuric type 1 diabetic patients. AB - AIM: To determine the long-term changes of the juxtaglomerular apparatus in incipient diabetic nephropathy. METHODS: Three renal needle biopsies were performed on 15 young type I diabetic patients with microalbuminuria; at baseline and after an average of 2.4 and 8.2 years. Using light microscopy, 1 microm serial sections of the plastic-embedded biopsies were investigated and volumes of the juxtaglomerular apparatus and glomerulus and areas of the macula densa and lumina of the afferent and efferent arterioles were measured. RESULTS: From baseline to second follow-up there was a significant decrease in JGA relative to glomerular volume. There was an increase in luminal area of the efferent arteriole which was paralleled by (non-significant) changes in the afferent arteriole. CONCLUSION: Over a period of 8.2 years JGA size remained stable, but decreased relative to glomerular size. Also, an increase in luminal area was noted in efferent arterioles. This may be due to increased single nephron blood flow secondary to nephron loss. PMID- 11900058 TI - Tumour necrosis factor-alpha (TNF), lymphotoxin and TNF receptor levels in serum from patients with Wegener's granulomatosis. AB - Wegener's granulomatosis (WG) is a systemic inflammatory disease with vasculitis as the key feature. Abnormal expression of tumour necrosis factor alpha (TNFalpha) is considered of prime pathogenic importance in several inflammatory diseases. The effects of TNFa are mediated by TNF receptors (TNF-R), and these receptors are often found in soluble forms (sTNF-R), which can modulate TNFalpha actions. To evaluate the clinical importance of the TNF family of cytokines, the serum levels of TNFalpha, TNFbeta, now termed lymphotoxin (LTalpha), and sTNF-R1 and sTNF-R2 were measured by ELISA in 8 patients with WG during active disease and during immunosuppressive treatment, and in 11 healthy controls in parallel. Serum concentrations of TNFalpha were undetectable in all except two controls (18%) and three patients with WG (37%). After 7 days of therapy, six of the WG patients had measurable TNFalpha levels. Examination of the relative amounts of TNFalpha and sTNF-R indicated that TNFalpha was mostly bound to its soluble receptors. In WG, the serum levels of sTNF-R1 and sTNF-R2 were dramatically increased (p<0.01), with little or no variation during treatment. While the IL 1beta levels did not deviate significantly from controls, the IL-1ra levels were significantly elevated in the WG patients throughout the study period (p<0.01). PMID- 11900060 TI - Mechanisms of resistance to imipenem in imipenem-resistant, ampicillin-sensitive Enterococcus faecium. AB - Enterococcus faecium has six penicillin-binding proteins (PBP), where PBP5 seems to be the main target for beta-lactam antibiotics. The PBP profiles of three imipenem-resistant, ampicillin-sensitive E. faecium strains, isolated from the same patient, were studied using biotinylated ampicillin and chemiluminescence detection. Imipenem resistance in these strains was found to be associated with hyperproduction of PBP5 compared to the ampicillin- and imipenem-susceptible strain ATCC 19434. PBP5 in the imipenem-resistant strains (S1, B2) exhibited a selectively decreased affinity for imipenem. An 854 bp DNA fragment, corresponding to the penicillin-binding domain of pbp5fm, was studied in the resistant strains and the reference strain. Four amino acid substitutions were observed in the resistant strains compared to the susceptible one. The contribution of these substitutions to the increased production of PBP5 in these strains is unclear since the substitution was observed also in a strain without increased production of PBP5. Our results suggest that the moderate imipenem resistance observed in these strains is associated with increased production of PBP5 with relatively decreased affinity for imipenem, and that evolution of imipenem resistance in E. faecium is dinstinct from that of the other beta lactams such as ampicillin. PMID- 11900059 TI - Comparative evaluation of a commercial test for rapid identification of methicillin-resistant Staphylococcus aureus. AB - The performance and ease of use of the recently introduced MRSA screen test (Denka Seiken Co. Ltd., Japan) for the identification of methicillin-resistant Staphylococcus aureus was evaluated in comparison with the BBL Crystal MRSA ID System (Becton Dickinson Europe, France). A total of 109 strains of S. aureus, consisting of 57 strains of MecA-negative S. aureus and 52 strains of MecA positive S. aureus, were tested. With MecA PCR as the gold standard, the MRSA screen test had 98% sensitivity and 98% specificity, whereas the BBL Crystal MRSA ID System had 98% sensitivity and 95% specificity. The simplicity of use and rapid result make the MRSA screen test a valuable tool in the clinical microbiology laboratory pending demonstration of the MecA gene that should still always be done. PMID- 11900061 TI - Multiple myeloma following an episode of community-acquired pneumococcal bacteraemia or meningitis. AB - The risk of multiple myeloma subsequent to an episode of serious pneumococcal infection has not been ascertained. We identified 328 episodes of community acquired pneumococcal bacteraemia and 77 episodes of pneumococcal meningitis in 227,000 persons over 40 years of age in the County of North Jutland, Denmark, in the period 1981 to 1996. The incidence rate of a subsequent diagnosis of multiple myeloma was determined by linkage to the Danish Cancer Registry. During 1,218 patient-years of follow-up in the bacteraemia cohort, 7 cases of multiple myeloma were diagnosed compared with 0.13 cases expected (standardized incidence ratio (SIR) 53.5, 95% confidence interval 21.4-111.4). During 444 patient-years of follow-up in the meningitis cohort, 4 cases of multiple myeloma were diagnosed compared with 0.05 cases expected (SIR 83.2, 95% confidence interval 22.6-214.8). Patients who survive an episode of community-acquired pneumococcal bacteraemia or meningitis are at increased risk of being diagnosed with multiple myeloma, but the absolute risk is low. PMID- 11900062 TI - Bond-market skepticism and stock-market exuberance in the hospital industry. AB - The hospital industry needs funds to refurbish physical facilities, upgrade clinical and information technologies, and rebuild financial positions weakened by past external challenges and unwise organizational strategies. The financial markets offer a marked contrast in capital access, as bond creditors remain skeptical while stock investors plunge back into the once-shunned industry. Ironically, high stock prices may drive the for-profit chains to repeat past cycles of overexpansion, while weak bond ratings may save non-profit systems from a comparable loss of focus on the core business of operating and improving inpatient facilities. This turbulence has implications for public payment, antitrust, and financial disclosure policies. PMID- 11900063 TI - The changing face of managed care. AB - Managed care plans--pressured by a variety of marketplace forces that have been intensifying over the past two years--are making important shifts in their overall business strategy. Plans are moving to offer less restrictive managed care products and product features that respond to consumers' and purchasers' demands for more choice and flexibility. In addition, because consumers and purchasers prefer broad and stable networks that require plans to include rather than exclude providers, plans are seeking less contentious contractual relationships with physicians and hospitals. Finally, to the extent that these changes erode their ability to control costs, plans are shifting from an emphasis only on increasing market share to a renewed emphasis on protecting profitability. Consequently, purchasers and consumers face escalating health care costs under these changing conditions. PMID- 11900064 TI - The financial health of California hospitals: a looming crisis. AB - This paper summarizes a Shattuck Hammond Partners study, released in late 2000, that found marked erosion in California hospitals' financial health. The study examined the revenue and expense dynamics that contributed to this erosion and explored future challenges, some of which are unique to California's regulatory environment, in the context of the hospital industry's current financial performance. The study concluded that California hospitals face a potential crisis--defined as the potential nonviability of a large portion of its hospital infrastructure--and explored the policy questions and implications of this situation. PMID- 11900065 TI - Out of the frying pan: New York City hospitals in an age of deregulation. AB - For several decades New York City hospitals had been distinguished by their tightly regulated environment, chronically weak finances, high occupancy rates, teaching intensity, dependency on public payers, low managed care penetration, and minimal merger activity. Then in the late 1990s a rapid convergence of forces -the Balanced Budget Act, managed care growth, state deregulation of commercial rates, escalating costs, and plunging hospital occupancy rates--threw the city's hospital industry into turmoil. In this paper we describe this period of turbulent change that has left most of the city's safety-net and small community hospitals near bankruptcy. PMID- 11900066 TI - Economic and demographic trends signal an impending physician shortage. AB - It is widely believed that the United States is producing too many physicians. We have approached this issue by developing a new model for workforce planning based on assessments of the macrotrends that underlie the supply and use of physician services. These trends include economic expansion, population growth, physicians' work effort, and the provision of services by nonphysician clinicians. Contrary to earlier predictions, this model projects that the United States soon will have a shortage of physicians and that if the pace of medical education remains unchanged, the shortage will become more severe. A dialogue focused on that eventuality is imperative. PMID- 11900067 TI - The ramifications of specialty-dominated medicine. PMID- 11900068 TI - Some thoughts on the white-follows-green law. PMID- 11900069 TI - A shortage of physicians or a surplus of assumptions? PMID- 11900070 TI - Through a different looking glass. PMID- 11900071 TI - Analyzing cause and effect in the U.S. physician workforce. PMID- 11900072 TI - Gauging supply and demand: the challenging quest to predict the future physician workforce. PMID- 11900073 TI - New opportunities for old mistakes. PMID- 11900074 TI - Inflation spurs health spending in 2000. PMID- 11900075 TI - Workers and their health plans: free to choose? PMID- 11900076 TI - Trends in health insurance coverage: a look at early 2001 data. PMID- 11900077 TI - Competitive behavior in the HMO marketplace. AB - Are health maintenance organizations (HMOs) less profitable in more competitive markets, and does competition erode unusually high profits over time? To answer these questions, we examined profit rates (as a proportion of revenues) in 1994 and 1997 for all HMOs in 259 metropolitan areas. We found that profits were significantly lower on average in 1994 in markets with more competition, measured alternatively by the number of HMOs or their market concentration. We also found that there was no relationship between a market's relative profit ranking in 1994 and its ranking in 1997; highly profitable markets were not able to preserve their relative standing. Neither the proportion of HMO enrollees in for-profit HMOs nor HMO market penetration was significantly related to profit rates. PMID- 11900078 TI - Physician and health system integration. AB - Incentives for vertical integration in the health care industry have led many hospitals to consolidate into health systems and profess a desire for closer alignment with affiliated physicians. In this study of fourteen organized delivery systems and their 11,000 physicians in sixty-nine medical groups, we found that many health systems did not align well with physicians. Even systems ostensibly committed to alignment emphasized structural relationships that did not enhance physician-system alignment and paid inadequate attention to issues of importance to physicians. This gap between the goal and reality of physician system alignment appears to be the result of systems' responding to a changing mix of policies, not all of which foster integration. PMID- 11900079 TI - Leaving nursing. PMID- 11900080 TI - Accountable but powerless. PMID- 11900082 TI - Bounceback: Blues thrive as markets cool toward HMOs. AB - Enrollment in Blue Cross and Blue Shield (BCBS) plans has grown by almost seventeen million since 1994, and recent financial performance indicators are positive for most plans in the Blues system. These gains have been achieved by for-profit, nonprofit, and mutually owned plans. A journalistic analysis of distinctive features contributing to recent successes is offered, combining observations of financial analysts, health services researchers, and BCBS officials. Long-term stability, broad provider networks, and conservative financial management have given the Blues advantages vis-a-vis many managed care organizations that have lost market share in the same period. PMID- 11900081 TI - Insuring low-income adults: does public coverage crowd out private? AB - During the mid-1990s Minnesota, Washington State, Oregon, and Tennessee implemented programs to provide subsidized health insurance for low-income persons who were not previously eligible for Medicaid. We estimate the effects of these programs on the health insurance status of low-income adults in these states. We find that among persons with family incomes below 100 percent of the federal poverty level, subsidized public coverage reduced the number of uninsured persons with very little effect on private coverage rates. Among persons with income between 100 percent and 200 percent of FPL, public coverage reduced the number of uninsured persons and crowded out some private insurance. The partial successes achieved by these programs should be kept in perspective: Even after program implementation, approximately 30 percent of low-income adults in the four states were uninsured. PMID- 11900083 TI - Geographic variation in the use of medications: is uniformity good news or bad? AB - Studies have repeatedly found much geographic variation in use of surgical and diagnostic procedures. This study of the variability of medication use for specific conditions in eleven California regions finds surprisingly few differences among regions. The difference between the highest- and lowest-use areas was far less than we anticipated and amounted to only 30-40 percent for many drugs. We explore five potential explanations for low geographic variability: financial incentives, impact of managed care, study design elements, characteristics of California, and pharmaceutical marketing and education efforts. To determine whether these findings represent good or bad news will require further study. PMID- 11900084 TI - The RWJF workers' compensation health initiative: findings and strategies. Robert Wood Johnson Foundation. PMID- 11900085 TI - Even playing field for consumer-directed long-term care. PMID- 11900087 TI - Care for the chronically ill requires bold changes. PMID- 11900086 TI - Consumer-directed care at home. PMID- 11900088 TI - We have the way, not the will. PMID- 11900089 TI - The long-term care burden. PMID- 11900090 TI - Dead armadillos. PMID- 11900091 TI - The economic value of medical technology. PMID- 11900092 TI - Medical technology's impacts and costs. PMID- 11900093 TI - Private health care in Australia. PMID- 11900094 TI - Integrating care: a talk with Kaiser Permanente's David Lawrence. Interview by Jeff Goldsmith. PMID- 11900095 TI - Defined-contribution health insurance products: development and prospects. AB - Defined-contribution health insurance products have received considerable recent attention, stimulated by double-digit increases in health plan premiums and employers' desire to get their employees more involved in health care purchasing decisions. Existing products typically feature a consumer health spending account, a major medical or other insurance policy, and the use of the Internet to support consumer decision making. They vary in their use of provider networks, provider payment approaches, the specific design of spending accounts, marketing strategies, and infrastructure investment. The companies producing these products are now at a critical juncture. They could grow rapidly over the next few years, be acquired by existing health plans, or fail if they do not deliver on their promises. PMID- 11900096 TI - Employer-sponsored health insurance: pressing problems, incremental changes. AB - Despite large premium increases, employers made only modest changes to health benefits in the past two years. By increasing copayments and deductibles and changing their pharmacy benefits, employers shifted costs to those who use services. Employers recognize these changes as short-term fixes, but most have not developed strategies for the future. Although interested in "defined contribution" benefits, employers do not agree about what this entails and have no plans for moving to defined contributions in the near future. While dramatic changes in health benefits are unlikely in the short term, policymakers may want to watch for future erosions in health coverage. PMID- 11900097 TI - Evolution in the Buyers Health Care Action Group purchasing initiative. AB - In 1997 the Buyers Health Care Action Group (BHCAG), a coalition of large employers in the Twin Cities, introduced a new purchasing initiative (called Choice Plus) designed to promote competition among care systems, driven by consumer choices. Our analysis suggests that consumers are playing the role, to some degree, envisioned by BHCAG. However, several issues now have caused BHCAG to dramatically restructure its approach to Choice Plus. It hopes that through this restructuring, Choice Plus will grow in the Twin Cities market and be adopted in other communities as well. The success of this new approach is by no means certain, as it faces a number of critical tests. PMID- 11900098 TI - Cost and quality trends in direct contracting arrangements. AB - This paper presents the first empirical analysis of a 1997 initiative of the Buyers Health Care Action Group (BHCAG) known as Choice Plus. This initiative entailed direct contracts with provider-controlled delivery systems; annual care system bidding; public reports of consumer satisfaction and quality; uniform benefits; and risk-adjusted payment. After case-mix adjustment, hospital costs decreased, ambulatory care costs rose modestly, and pharmacy costs increased substantially. Process-oriented quality indicators were stable or improved. The BHCAG employer-to-provider direct contracting and consumer choice model appeared to perform reasonably well in containing costs, without measurable adverse effects on quality. PMID- 11900099 TI - Influences of intentional and unintentional forgetting on false memories. AB - In 2 experiments, we examined the interplay of 2 types of memory errors: forgetting and false memory--errors of omission and commission, respectively. We examined the effects of 2 manipulations known to inhibit retrieval of studied words--directed forgetting and part-list cuing--on the false recall of an unstudied "critical" word following study of its 15 strongest associates. Participants cued to forget the 1st of 2 studied lists before studying the 2nd recalled fewer List 1 words but intruded the missing critical word more often than did participants cued to remember both lists. By contrast, providing some studied words as cues during recall reduced both recall of the remaining studied words and intrusions of the critical word. The results suggest that forgetting can increase or decrease false memories, depending on whether such forgetting reflects impaired access to an entire episode or retrieval competition among elements of an episode. PMID- 11900100 TI - Detecting variety: what's so special about uniformity? AB - People and pigeons were taught to make 1 of 2 responses to 16-icon arrays that differed in their visual variability. In 2 experiments, participants had to (a) discriminate a collection of identical items from a collection in which 2 or more items were different or (b) discriminate a collection of different items from a collection in which 2 or more items were identical. In Experiment 1, humans found it much easier to discriminate uniformity from all levels of diversity. In Experiment 2, pigeons also found it easier to discriminate uniformity from all levels of diversity, but the size of this effect was smaller than that observed in people. These and other results suggest that both species are predisposed to notice differences rather than similarities. PMID- 11900101 TI - Prototypes and particulars: geometric and experience-dependent spatial categories. AB - People use geometric cues to form spatial categories. This study investigated whether people also use the spatial distribution of exemplars. Adults pointed to remembered locations on a tabletop. In Experiment 1, a target was placed in each geometric category, and the location of targets was varied. Adults' responses were biased away from a midline category boundary toward geometric prototypes located at the centers of left and right categories. Experiment 2 showed that prototype effects were not influenced by cross-category interactions. In Experiment 3, subsets of targets were positioned at different locations within each category. When prototype effects were removed, there was a bias toward the center of the exemplar distribution, suggesting that common categorization processes operate across spatial and object domains. PMID- 11900102 TI - Binding in short-term visual memory. AB - The integration of complex information in working memory, and its effect on capacity, shape the limits of conscious cognition. The literature conflicts on whether short-term visual memory represents information as integrated objects. A change-detection paradigm using objects defined by color with location or shape was used to investigate binding in short-term visual memory. Results showed that features from the same dimension compete for capacity, whereas features from different dimensions can be stored in parallel. Binding between these features can occur, but focused attention is required to create and maintain the binding over time, and this integrated format is vulnerable to interference. In the proposed model, working memory capacity is limited both by the independent capacity of simple feature stores and by demands on attention networks that integrate this distributed information into complex but unified thought objects. PMID- 11900103 TI - Making a silk purse out of two sow's ears: young children's use of comparison in category learning. AB - Comparison mechanisms have been implicated in the development of abstract, relational thought, including object categorization. D. Gentner and L. L. Namy (1999) found that comparing 2 perceptually similar category members yielded taxonomic categorization, whereas viewing a single member of the target category elicited shallower perceptual responding. The present experiments tested 2 predictions that follow from Gentner and Namy's (1999) model: (a) Comparison facilitates categorization only when the targets to be compared share relational commonalities, and (b) providing common labels for targets invites comparison, whereas providing conflicting labels deters it. Four-year-olds participated in a forced-choice task. They viewed 2 perceptually similar target objects and were asked to "find another one." Results suggest an important role for comparison in lexical and conceptual development. PMID- 11900105 TI - Strategic regulation of grain size in memory reporting. AB - To increase their report accuracy, rememberers may either withhold information that they feel unsure about or provide relatively coarse information that is unlikely to be wrong. In previous work (A. Koriat & M. Goldsmith, 1996c), the authors delineated the metacognitive monitoring and control processes underlying the decision to volunteer or withhold particular items of information (report option) and examined how these processes are used in the strategic regulation of memory accuracy. This article adapts that framework to address control over the grain size (precision-coarseness) of the information that people report. Results show that rememberers strategically regulate the grain of their answers to accommodate the competing goals of accuracy and informativeness. The metacognitive processes underlying this regulation are elucidated. PMID- 11900104 TI - Functional MRI evidence for an abstract, not perceptual, word-form area. AB - Previous studies have found an area in left ventral visual cortex that responds more to words and pseudowords than to consonant strings. Does this area respond to the perceptual form of wordlike stimuli, or is it responding to some more abstract, linguistic property, such as orthographic regularity (i.e., conformity with the spelling rules of the language)? During a functional magnetic resonance imaging experiment, participants read alternating-case words and pseudowords, which are orthographically regular but are perceptually unfamiliar. These stimuli activated the same area that was activated by pure-case words and pseudowords. These results suggest that the response of the so-called word-form area is not based on perceptual familiarity but rather on some more abstract feature such as orthographic regularity. PMID- 11900106 TI - The specificity of maternal disclosure of HIV/AIDS in relation to children's adjustment. AB - Disclosure experience of 58 HIV-seropositive women was examined as a multi faceted process comprising eight variables: level, seriousness, breadth, frequency, source, secrecy, age at first disclosure, and time since disclosure. The majority of children (57%) were told that their mothers had HIV/AIDS and were given additional information about mothers' health (64%) including prognosis of potential death (68%). Most were disclosed to by their mothers (75%) and were not asked to keep disclosures secret (66%). For most (68%) discussion regarding mother's health was infrequent. Children, on average, were first disclosed information at age 7 and had been aware of information for 3 years. Disclosure characteristics were related to demographics of mothers and children. Hierarchical multiple regression analyses showed that children asked to keep disclosures secret tended to display more behavior problems than children not asked to keep secrets. However, the specificity of disclosure did not otherwise predict children's adaptive functioning. PMID- 11900107 TI - HIV-positive notification and behavior changes in Montreal injection drug users. AB - This study examined the effect of an HIV-positive notification on the sexual and injection behaviors, living conditions, and medical demands of injection drug users (IDUs) in Montreal. The behavior changes of 73 IDUs who received an HIV positive test result, and 219 IDUs who tested negative were compared, and the net difference (ND) in the proportion of IDUs adopting the particular change was determined. No significant changes were found in drug use or needle sharing practices. A higher proportion of IDUs who received an HIV-positive notification acquired unstable living conditions, ND = 20.7% (95% CI = 3.3, 38.1), began medical follow up, ND = 34.4% (95% CI = 20.8, 48.7), and increased needle exchange program (NEP) utilization, ND = 20.5% (95% CI = 8.3, 32.8). Compared with HIV-negative males, more HIV-positive male IDUs stopped sexual relations, ND = 24.6% (95% CI = 0.4, 48.9), and sex work, ND = 31.8% (95% CI = 12.4, 51.3), and fewer began new relations, ND = -38.2% (95% CI = -52.6, -23.9). The medical community and NEPs have an important role in providing support for newly diagnosed IDUs. PMID- 11900109 TI - Risk among men who have sex with men in the United States: a comparison of an Internet sample and a conventional outreach sample. AB - This study compared the demographics and risk behaviors of two samples of men who have sex with men (MSM), using cross-sectional data that were collected via the Internet and through conventional bar-based outreach. The Internet sample was significantly older, more likely to identify as "bisexual," and less educated than the bar sample. After controlling for age and education, few differences were observed between the samples. However, three variables that markedly differentiated the samples were history of sexually transmitted disease infection, HIV serostatus, and sources utilized to obtain health information. No difference in Internet use was found. Based on the possible decreased social desirability promoted by the use of electronic data collection methodologies, these findings provide preliminary evidence that Internet and bar respondents are similar and that the Internet may serve as an expedient as well as reliable methodology to increase understanding of risk among MSM. PMID- 11900108 TI - Effects of fatalism and family communication on HIV/AIDS awareness variations in Native American and Anglo parents and children. AB - Incorporating three analytic approaches, the present research examines HIV/AIDS awareness levels in a sample of Native American and Anglo parents and children. Descriptive analysis revealed that Native Americans, especially children, possess startlingly poor levels of HIV/AIDS-related knowledge compared with Anglos. This disparity is most evident for more subtle HIV/AIDS facts. Using an all-or-none scoring model disclosed even larger ethnic discrepancies. Multiple regression analysis demonstrated that fatalism and family communication mediate the HIV/AIDS awareness-ethnicity relationship beyond the effects of socioeconomic status. PMID- 11900110 TI - Internet use among people living with HIV/AIDS: association of health information, health behaviors, and health status. AB - The telecommunications revolution provides open access to health information that can inform and empower people living with chronic illnesses. However, many people living with HIV may not access the Internet and are not benefiting from available health information. This study investigated Internet access among people living with HIV/AIDS and its relation to health. Results of a survey of men (n = 175) and women (n = 84) living with HIV/AIDS recruited from infectious disease clinics and community-based AIDS services showed that 51% (n = 116) of participants reported ever using the Internet, of which 59% (n = 68) had used the Internet to access health-related information. As expected, Internet users were significantly more likely to be better educated and of higher incomes. Internet users, including those who used the Internet for general purposes and those who reported health-related use, also demonstrated significantly greater knowledge of HIV disease and greater confidence in their ability to adhere to medications. Persons who used the Internet for general purposes were more likely to have an undetectable viral load compared to persons who had not used the Internet. The disparities in Internet use identified in this study suggest that individuals who access the Internet, particularly for health information, are among the better resourced and healthier persons living with HIV/AIDS. PMID- 11900111 TI - Understanding gender differences in condom use self-efficacy among youth in urban Cameroon. AB - The aim of this study was to understand gender differences in components of condom use self-efficacy to inform the design of effective reproductive health interventions for youth. Data stem from a July to August 2000 reproductive health survey among youth aged 15-24. Gender differentials in self-efficacy are analyzed using logistic regression. Perceived ability to discuss and negotiate condom use is high for both sexes. Women are less likely than men to know correct condom use (58% vs. 80%, p<.01) but are more likely to be shy buying condoms (67% vs. 50%, p<.01). Prior experience buying and using condoms, parental support, and condom promotion affect perceived ability to correctly use condoms and shyness buying condoms. Programs aiming to increase self-efficacy in condom use should focus on increasing confidence in youth's ability to buy condoms and to use them correctly, especially for young women. There is a need for programs that publicize and/or increase access to youth-friendly outlets and increase the acceptability of young women buying condoms. PMID- 11900112 TI - Correlates of HIV risk among Ecuadorian adolescents. AB - The purpose of this study was to examine the relationship of HIV knowledge, demographics, and psychosocial factors with HIV risk behavior among high school students (N = 805) in urban and rural regions of Ecuador. Forty-three percent of the participants reported being sexually experienced. Of the sexually experienced participants, 50% reported never using condoms for intercourse, and 70% did not use condoms at last intercourse. A small minority of the sample felt that they were at risk for contracting HIV (18.5%). Educational aspirations past high school and residence in an urban area were significantly associated with decreased HIV risk (condom use at last intercourse and abstinence). High self efficacy for condom use and strong refusal skills to unsafe sex were also significantly associated with decreased HIV risk. Consistent with prior research, the participants that reported their peers were sexually experienced were significantly more likely to have had sex. The discussion focuses on the use of interventions and education for promoting safer sexual behavior among Ecuadorian adolescents. PMID- 11900113 TI - Production and characterization of pullulan from beet molasses using a nonpigmented strain of Aureobasidium pullulans in batch culture. AB - The production of pullulan from beet molasses by a pigment-free strain of Aztreobasidium pullulans on shake-flask culture was investigated. Combined pretreatment of molasses with sulfuric acid and activated carbon to remove potential fermentation inhibitors present in molasses resulted in a maximum pullulan concentration of 24 g/L, a biomass dry wt of 14 g/L, a pullulan yield of 52.5%, and a sugar utilization of 92% with optimum fermentation conditions (initial sugar concentration of 50 g/L and initial pH of 7.0). The addition of other nutrients as carbon and nitrogen supplements (olive oil, ammonium sulfate, yeast extract) did not further improve the production of the exopolysaccharides. Structural characterization of the isolated polysaccharides from the fermentation broths by 13C-nuclear magnetic resonance spectroscopy and pullulanase digestion combined with size-exclusion chromatography confirmed the identity of pullulan and the homogeneity (>93% dry basis) of the elaborated polysaccharides by the microorganism. Using multiangle laser light scattering and refractive index detectors in conjunction with high-performance size-exclusion chromatography molecular size distributions and estimates of the molecular weight (Mw = 2.1-4.1 x 10(5)), root mean square of the radius of gyration (R = 30-38 nm), and polydispersity index (Mw/Mn = 1.4-2.4) were obtained. The fermentation products of molasses pretreated with sulfuric acid and/or activated carbon were more homogeneous and free of contaminating proteins. In the concentration range of 2.8 10.0 (w/v), the solution's rheologic behavior of the isolated pullulans was almost Newtonian (within 1 and 1200 s(-1) at 20 degrees C); a slight shear thinning was observed at 10.0 (w/v) for the high molecular weight samples. Overall, beet molasses pretreated with sulfuric acid and activated carbon appears as an attractive fermentation medium for the production of pullulan by A. pullulans. PMID- 11900114 TI - Cloning, expression, and characterization of thermostable region of amylopullulanase gene from Thermoanaerobacter ethanolicus 39E. AB - The bifunctional activities of alpha-amylase and pullulanase are found in the cloned recombinant amylopullulanase. It was encoded in a 2.9-kb DNA fragment that was amplified using polymerase chain reaction from the chromosomal DNA of Thermoanaerobacter ethanolicus 39E. An estimated 109-kDa recombinant protein was obtained from the cloned gene under the prokaryotic expression system. The optimum pH of the recombinant amylopullulanase was 6.0. The most stable pH for the alpha-amylase and pullulanase activity was 5.5 and 5.0, respectively. The optimum temperature for the alpha-amylase activity was 90 degrees C, while its most stable temperature was 80 degrees C. Regarding pullulanase activity, the optimum temperature and its most stable temperature were found to be 80 and 75 degrees C, respectively. Pullulan was found to be the best substrate for the enzyme. The enzyme was activated and stabilized by the presence of Ca2+, whereas EDTA, N-bromosuccinimide, and alpha-cyclodextrin inhibited its bifunctional activities. A malto-2-4-oligosaccharide was the major product obtained from the enzymatic reaction on soluble starch, amylose, amylopectin, and glycogen. A single maltotriose product was found in the pullulan hydrolysis reaction using this recombinant amylopullulanase. Kinetic analysis of the enzyme indicated that the Km values of alpha-amylase and pullulanase were 1.38 and 3.79 mg/mL, respectively, while the Vmax values were 39 and 98 micromol/(min x mg of protein), respectively. PMID- 11900115 TI - Archaeal tetraether lipids: unique structures and applications. AB - The extremely stable biomolecules manufactured by organisms from extreme environments are of great scientific and engineering interest in the development of robust and stable industrial biocatalysts. Identification of molecules that impart stability under extremes will also have a profound impact on our understanding of cellular survival. This review discusses isolation and characterization of archaeal tetraethers as well as target technologies for tetraether lipid application. The isolation and characterization of archaeal tetraether lipids has led to some interesting applications improving on ester lipid technologies. Potential applications include novel lubricants, gene delivery systems, monolayer lipid matrices for sensor devices, and protein stabilization. Following this review, patent abstracts and additional literature pertaining to the isolation, characterization, and application of archaeal membrane lipids are listed. PMID- 11900116 TI - Rashes among schoolchildren--14 states, October 4, 2001-February 27, 2002. AB - Fourteen states (Arizona, Connecticut, Florida, Georgia, Indiana, Mississippi, New York, Ohio, Oregon, Pennsylvania, Texas, Virginia, Washington, and West Virginia) have reported investigations of multiple schoolchildren who have developed rashes. This report summarizes the investigation by state and local health departments of these rashes, which have occurred during October 2001 through February 2002, and provides examples for four states. Preliminary findings indicate that further investigation is needed to determine whether a common etiology for these rashes exists. PMID- 11900117 TI - Congenital malaria as a result of Plasmodium malariae--North Carolina, 2000. AB - Congenitally acquired malaria is rare in the United States; < or = 10 cases are reported each year. Congenital infection with Plasmodium malariae is particularly uncommon because distribution of this parasite is focal and sparse in areas where P. falciparum is endemic. The last case of congenital P. malariae infection in the United States was reported in 1992. This report describes the investigation of a case of P. malariae in an infant with no travel history outside of the United States and suggests that health-care providers suspect malaria when treating a neonate or young infant with fever if the mother has traveled or lived in a malarious area. PMID- 11900118 TI - Health-related quality of life--Puerto Rico, 1996-2000. AB - Although a number of studies have been made to determine the health-related quality of life (HRQOL) of persons living in the United States, no overall assessment of HRQOL has been conducted previously for residents of Puerto Rico. To determine the HRQOL of adults living in Puerto Rico, during 1996-2000, as part of the Behavioral Risk Factor Surveillance System (BRFSS), interviews were conducted in Spanish with a representative sample of Puerto Rican adults. Older women, persons with less education or lower income, persons unable to work, and those who were overweight or who had diabetes or high blood pressure reported more days for which they were physically or mentally unhealthy during the 30 days preceding the survey. Interventions designed to reach these vulnerable, demographic, socioeconomic, and behavioral risk groups might help adults in Puerto Rico increase their quality and years of healthy life and eliminate health disparities. PMID- 11900119 TI - Evaluation of the performance of Hartmann sensors in strong scintillation. AB - A simulation study is presented that evaluates the performance of Hartmann wave front sensors with measurements obtained with the Fried geometry and the Hutchin geometry. Performance is defined in terms of the Strehl ratio achieved when the estimate of the complex field obtained from reconstruction is used to correct the distorted wave front presented to the wave-front sensor. A series of evaluations is performed to identify the strengths and the weaknesses of Hartmann sensors used in each of the two geometries in the two-dimensional space of the Fried parameter r0 and the Rytov parameter. We found that the performance of Hartmann sensors degrades severely when the Rytov number exceeds 0.2 and the ratio l/r0 exceeds 1/4 (where l is the subaperture side length) because of the presence of branch points in the phase function and the effect of amplitude scintillation on the measurement values produced by the Hartmann sensor. PMID- 11900120 TI - One-parameter two-term Henyey-Greenstein phase function for light scattering in seawater. AB - A one-parameter two-term Henyey-Greenstein (TTHG) phase function of light scattering in seawater is proposed. The original three-parameter TTHG phase function was reduced to the one-parameter TTHG phase function by use of experimentally derived regression dependencies between integral parameters of the marine phase functions. An approach to calculate a diffuse attenuation coefficient in the depth of seawater is presented. PMID- 11900121 TI - Apodization effects in the retrieval of volume mixing ratio profiles. AB - In remote sensing applications, spectra measured by Fourier-transform spectrometers are routinely apodized. A rigorous analysis approach would explicitly account for correlations induced in the covariance matrix by apodization, but these correlations are often ignored to simplify and speed up the processing. Using spectra measured by the Atmospheric Trace Molecule Spectroscopy missions, we investigated the effect of apodization on the retrieval of volume mixing ratio profiles for the case in which these correlations are ignored. Minor discrepancies occur between results for apodized and unapodized spectra, particularly when lines with a low signal-to-noise ratio are fitted. A set of microwindows is reported for O3 in the range of 1550-3350 cm(-1). PMID- 11900122 TI - Phase function effects on oceanic light fields. AB - Numerical simulations show that underwater radiances, irradiances, and reflectances are sensitive to the shape of the scattering phase function at intermediate and large scattering angles, although the exact shape of the phase function in the backscatter directions (for a given backscatter fraction) is not critical if errors of the order of 10% are acceptable. We present an algorithm for generating depth- and wavelength-dependent Fournier-Forand phase functions having any desired backscatter fraction. Modeling of a comprehensive data set of measured inherent optical properties and radiometric variables shows that use of phase functions with the correct backscatter fraction and overall shape is crucial to achieve model-data closure. PMID- 11900123 TI - Fabrication of advanced fiber Bragg gratings by use of sequential writing with a continuous-wave ultraviolet laser source. AB - We present a novel scheme based on sequential writing for fabrication of advanced fiber Bragg gratings. As opposed to earlier sequential methods this technique uses a cw UV laser source and allows for very precise control and repetitivity of the formation of the gratings. Furthermore it is possible to use high average irradiances without destroying the fiber, resulting in considerable reduction in fabrication time for complex gratings. The method has been applied to several test gratings, which proved its versatility and quality. PMID- 11900124 TI - Optimal dispersion precompensation by pulse chirping. AB - For the procedure of dispersion precompensation in fibers by prechirping, we found that there is a maximum distance over which a pulse initially compressed by prechirping can return to its original width. The distance constraint comes in the form of a mathematical relationship involving the distance, dispersion, initial pulse width, and peak power, implying that the restriction governs all the fiber parameters. Simple closed-form approximations for the constraint and for the corresponding required prechirp are derived on the basis of a variational approach. The validity of the analytical formulas is confirmed by split-step Fourier numerical simulation. PMID- 11900125 TI - Optical design and evaluation of a three-dimensional imaging and ranging system based on time-correlated single-photon counting. AB - The design and operation of a noncontact surface profilometry system based on the time-correlated single-photon-counting technique are described. This system has a robust optomechanical design and uses an eye-safe laser that makes it particularly suitable for operation in an uncontrolled industrial environment. The sensitivity of the photon-counting technique permits its use on a variety of target materials, and its mode of operation does not require the continual presence of an operator. The system described has been optimized for a 1-25-m standoff, has a distance repeatability of <30 microm, and has a transverse spatial resolution of approximately 60 microm at a 2-m standoff and approximately 400 microm at a 13-m standoff. PMID- 11900126 TI - Polarization characteristic of a room-temperature Co:MgF2 laser. AB - A study of the polarization characteristic of a Co:MgF2 laser with a 1320-nm YAG pumping laser at room temperature is reported. The thresholds, output energies, and efficiencies of the laser are given at the various polarization states. The more intensive emission is in the pi-polarization pump laser and sigma polarization laser operation. Performances of the Co:MgF2 lasers are similar for the polarized and unpolarized laser pumping along the optical axis of the crystal. PMID- 11900127 TI - Passively Q-switched quasi-three-level laser and its intracavity frequency doubling. AB - A passively Q-switched quasi-three-level Nd:YAG laser is intracavity frequency doubled to generate a blue laser. The 473-nm blue laser has a peak power of 37 W and a pulse width of 23 ns at a pumping power of 1.6 W. To model this laser numerically, we developed rate equations by taking into consideration both the quasi-three-level nature of the gain medium and the four-level nature of the saturable absorber. Good agreement was achieved between experimental and simulated results for both the fundamental and the second-harmonic output. The reabsorption loss of the gain medium is estimated under pulsed operation. PMID- 11900128 TI - Output power and polarization characteristics for a diode-side-pumped Nd:YAG rod laser with a diffusive optical pump cavity. AB - We fabricated and analyzed the output power and polarization characteristics of an efficient diode-side-pumped Nd:YAG rod laser with a diffusive optical cavity. The resonator stability conditions are analyzed graphically in the symmetric and asymmetric configurations for a plane-parallel resonator. On the basis of an analysis of the stability condition and mode size for the r and theta polarizations, we clarify how the stable laser operation is possible for various resonator configurations. In particular, we show that the critical stability region of around g1*g2* = 0 provides a stable resonator in the symmetric resonator, even with a slight asymmetry. Experimentally, the output power and polarization characteristics are confirmed in association with the resonator stability condition. PMID- 11900129 TI - Design and fabrication of a diode-side-pumped Nd:YAG laser with a diffusive optical cavity for 500-W output power. AB - A ray-tracing code has been developed, and the design parameters of the laser pump head were analyzed in terms of crystal diameter, doping concentration, and optical cavity diameter. According to the numerical analysis, we fabricated an efficient diode-pumped Nd:YAG laser and experimentally obtained 500-W output power. The output power is close to the numerically calculated output power of approximately 450 W and corresponds to an optical-to-optical conversion efficiency of 46.7% and an optical slope efficiency of 49%. PMID- 11900130 TI - Cost-effective low timing jitter passively Q-switched diode-pumped solid-state laser with composite pumping pulses. AB - A novel scheme that combines gain switching with passive Q switching of a miniature diode-pumped solid-state laser is proposed and implemented. A composite pumping pulse, consisting of a long, low-intensity pulse and a following short, high-intensity pulse, is used to reduce the timing jitter. A greater-than-tenfold reduction in timing jitter is demonstrated. PMID- 11900131 TI - Asynchronously modulated waves in a ring laser cavity. AB - When modulated through the harmonic motion of one mirror, the counterpropagating waves in a ring laser oscillate out of phase. A solution to the wave equation is presented that satisfies both the time-dependent boundary condition and the resonance condition. This theoretical prediction is confirmed experimentally to leading order in terms that are inversely proportional to the speed of light. The method of solution is applicable to arbitrary phase modulation at more than one spatial location in the cavity. Potential uses include the reduction of the locking problem in ring lasers and the testing of higher-order kinematic effects in the theory of relativity. PMID- 11900133 TI - Theoretical analysis of a noncollinear phase-matched optical parametric amplifier seeded by an optical parametric generation. AB - We have theoretically analyzed the characteristics of an optical parametric amplifier system seeded by an optical parametric generation. We investigated the influences of the energy, pulse duration, material dispersion, and the third order nonlinear effect in beta-barium borate. The group-velocity mismatch (GVM) becomes the most important factor for the amplification of bandwidths. Even though tilting the wave front of the pump can decrease the GVM, it seems impossible to generate pulses smaller than 10-fs with 400-nm pumping. However, 10 fs pulses can be achieved with a 30-fs pump duration with pumping at 800 nm. PMID- 11900132 TI - Viscosity dependence of optical limiting in carbon black suspensions. AB - We measure the optical limiting behavior of carbon black suspensions in various viscosity solvents by using a 10-Hz repetition rate, 532-nm, 5-ns pulsed laser. We found that, for common solvents used in the past such as water and ethanol, the limiting behavior ceases after a few laser firings and a turnover in the limiting curve appears. This can be explained by depletion of the carbon black within the focal volume. This turnover shifts to lower energies as the viscosity of the solvent becomes greater. However, for low viscosity liquids, such as carbon disulfide or pentane, the limiting is unaffected by the repetition rate, at least for frequencies up to 10 Hz, because of diffusion of the carbon black particles. This diffusion allows fresh material to replace the irradiated volume within the time between pulses. PMID- 11900134 TI - Reduction of the temporal fluctuation of the signal intensity in optical wave mixing with a reflection grating. AB - Reduction of the temporal fluctuation of the signal intensity in an optical wave mixing system with a reflection grating is studied by use of a feedback of the output pump with appropriate feedback reflectivity. The advantage of the method is that the temporal fluctuation of the output intensity can be significantly reduced, although its mean intensity is not reduced. PMID- 11900135 TI - Upconversion fluorescence and optical power limiting effects based on the two- and three-photon absorption process of a new organic dye BPAS. AB - A new organic dye, trans-4, 4'-bis(pyrrolidingl) stilbene (BPAS), with large two photon absorption (TPA) and three-photon absorption (3PA) has been synthesized. The molecular TPA cross section sigma2' at 550-670 nm and the 3PA cross section sigma3' at 720-1000 nm have been measured. The biggest sigma2' and sigma3' was 5.77 x 10(-47) cm4s/photon and 27 x 10(-75) cm6 s2 at 600 and 980 nm, respectively. In the experiment process we found that the strongest TPA wavelength is not at two times of the strongest linear absorption wavelength, but there is some blue shift. The 3PA-induced optical power-limiting properties have also been illustrated at 980 nm. PMID- 11900136 TI - Tunable continuous-wave doubly resonant optical parametric oscillator by use of a semimonolithic KTP crystal. AB - A temperature-tuned continuous-wave doubly resonant optical parametric oscillator (OPO) consisting of a semimonolithic KTP crystal and a concave mirror has been designed and built. Under single-axial-mode-pair operation, we obtained a combined output power of the signal and idler light fields up to 365 mW at a pump power of 680 mW. The output wavelength of the OPO can be temperature tuned by as much as 9 nm. We achieved 2.8-GHz continuous frequency tuning of the OPO by tuning the pump laser frequency. PMID- 11900137 TI - Mathematical formulations for the schlieren detection method applied to the measurement of photodeformation. AB - In a schlieren detection scheme for photodeformation measurements, the divergence of the probe beam that is induced by the axisymmetric but radially inhomogeneous periodic photothermal displacement of the surface of a sample is transformed into an intensity variation by insertion of an iris in front of the detection photodiode. We present three expressions for the intensity profile of a Gaussian laser beam that is reflected by the inhomogeneous photodeformation of a solid. The first expression proceeds from geometrical optics (or photometry), whereas the second one derives from the use of the well-known ABCD law and the third one from diffraction principles. Comparing these formulations of the schlieren signal with their behavior as a function of different geometrical parameters, we obtain the domain of validity of each expression, and we deduce the advantages of the different formalisms. PMID- 11900139 TI - Analytical differentiation of the differential-absorption-lidar data distorted by noise. AB - A method of analytical differentiation is developed for processing differential absorption lidar (DIAL) data. The method is based on simple analytical transformation of the DIAL on and off signal ratio. The derivatives consequently are found for either individual data points or local zones of the measurement range. The method makes possible the separation of local zones of interest and the separate investigation of these. The smoothing level is established by the selected value of the exponent in a transformation formula rather than by the selection of the resolution range. The method does not require the calculation of local signal increments. This reduces significantly the high-frequency noise in the measured concentration. The method is general and can be used for different experimental data, including inelastic (Raman) lidar data. The processing technique is practical and does not require a determination of the solution for a large set of algebraic equations. It is based on the simple repetition of the same type of calculations with different constants. The method can easily be implemented for practical computations. PMID- 11900138 TI - Two-channel direct-detection Doppler lidar employing a charge-coupled device as a detector. AB - A direct-detection Doppler lidar system is demonstrated that uses a CCD as a detector for the first time to our knowledge. The ability to use this linear device with the circular output from a Fabry-Perot etalon comes from use of a circle-to-line converter [Appl. Opt. 29, 1482 (1990)]. In addition to the gains in quantum efficiency obtained through use of this detector, the lidar system described in this paper also has the capability to measure winds from aerosol and molecular backscatter simultaneously in two separate channels by directing the light reflected from one channel into the other. Early measurements with this system are presented; it is shown that, although accurate aerosol wind measurements are easily obtained, molecular measurements require a carefully calibrated inverse model and special hardware to derive accurate wind measurements with this channel. PMID- 11900140 TI - Single-mode, tunable output from a midwave-infrared-seeded optical parametric oscillator system. AB - We have built a coupled master oscillator and a slave oscillator optical parametric oscillator (OPO) system that provides single-frequency, pulsed radiation in the midwave infrared (MWIR). The direct-diode-pumped master OPO provided narrow-band (<6-MHz, instrument-limited), tunable MWIR cw radiation that was used to seed a higher-peak-power pulsed slave OPO. When seeded directly at the MWIR idler, the pulsed output of the slave OPO was constrained to oscillate on a single longitudinal mode even though the slave OPO is pumped by a multilongitudinal-mode laser source. The linewidth of the pulsed output has been measured to be <220 MHz (instrument limited), which is well suited for coherent differential absorption lidar applications. PMID- 11900141 TI - Thermoelectrically cooled quantum-cascade-laser-based sensor for the continuous monitoring of ambient atmospheric carbon monoxide. AB - We report the first application of a thermoelectrically cooled, distributed feedback quantum-cascade laser for continuous spectroscopic monitoring of CO in ambient air at a wavelength of 4.6 microm. A noise-equivalent detection limit of 12 parts per billion was demonstrated experimentally with a 102-cm optical pathlength and a 2.5-min data acquisition time at a 10-kHz pulsed-laser repetition rate. This sensitivity corresponds to a standard error in fractional absorbance of 3 x 10(-5). PMID- 11900142 TI - Experiments on light scattering and extinction by small, micrometer-sized aggregates of spheres. AB - We performed experiments to study the extinction, scattering, and polarization of light by ensembles of fractal dust aggregates that consist of spherical monomers large compared with the wavelength. Extinction was measured on a homogeneous dust cloud. Scattering and polarization were measured on a collimated dust beam. We found that polarization and extinction are determined only by a small size scale that is defined by a monomer and its closest neighbors in an aggregate. The scattering function might also depend on the overall size of the aggregate or the total number of monomers in an aggregate. PMID- 11900143 TI - Computation of a spectrum from a single-beam fourier-transform infrared interferogram. AB - A new high-accuracy method has been developed to transform asymmetric single sided interferograms into spectra. We used a fraction (short, double-sided) of the recorded interferogram and applied an iterative correction to the complete recorded interferogram for the linear part of the phase induced by the various optical elements. Iterative phase correction enhanced the symmetry in the recorded interferogram. We constructed a symmetric double-sided interferogram and followed the Mertz procedure [Infrared Phys. 7,17 (1967)] but with symmetric apodization windows and with a nonlinear phase correction deduced from this double-sided interferogram. In comparing the solution spectrum with the source spectrum we applied the Rayleigh resolution criterion with a Gaussian instrument line shape. The accuracy of the solution is excellent, ranging from better than 0.1% for a blackbody spectrum to a few percent for a complicated atmospheric radiance spectrum. PMID- 11900144 TI - Linear excitation schemes for IR planar-induced fluorescence imaging of CO and CO2. AB - A detailed discussion of linear excitation schemes for IR planar-induced fluorescence (PLIF) imaging of CO and CO2 is presented. These excitation schemes are designed to avoid laser scattering, absorption interferences, and background luminosity while an easily interpreted PLIF signal is generated. The output of a tunable optical parametric amplifier excites combination or overtone transitions in these species, and InSb IR cameras collect fluorescence from fundamental transitions. An analysis of the dynamics of pulsed laser excitation demonstrates that rotational energy transfer is prominent; hence the excitation remains in the linear regime, and standard PLIF postprocessing techniques may be used to correct for laser sheet inhomogeneities. Analysis of the vibrational energy-transfer processes for CO show that microsecond-scale integration times effectively freeze the vibrational populations, and the fluorescence quantum yield following nanosecond-pulse excitation can be made nearly independent of the collisional environment. Sensitivity calculations show that the single-shot imaging of nascent CO in flames is possible. Signal interpretation for CO2 is more complicated, owing to strongly temperature-dependent absorption cross sections and strongly collider-dependent fluorescence quantum yield. These complications limit linear CO2 IR PLIF imaging schemes to qualitative visualization but indicate that increased signal level and improved quantitative accuracy can be achieved through consideration of laser-saturated excitation schemes. PMID- 11900145 TI - Simulation of uplooking and downlooking high-resolution radiance spectra with two different radiative transfer models. AB - Measurements of up-looking spectral radiances measured during the Convection and Moisture Experiment and down-looking spectral radiances measured at one of the Atmospheric Radiation Measurement sites are compared with simulations with use of two different line-by-line models. Simulations are performed in tightly controlled conditions to verify the behavior of the models. Spectra computed at higher samplings are used to study the spectral structure of the differences between simulations and measurements. A revised list of water vapor spectroscopic parameters is used to test the impact of improved spectroscopic data on the accuracy of the line-by-line calculations. The sensitivity of the results to errors that result from uncertainties in the input atmospheric temperature and humidity profiles is also investigated. PMID- 11900146 TI - Measurement of seeing and the atmospheric time constant by differential scintillations. AB - A simple differential analysis of stellar scintillations measured simultaneously with two apertures opens the possibility to estimate seeing. Moreover, some information on the vertical turbulence distribution can be obtained. A general expression for the differential scintillation index for apertures of arbitrary shape and for finite exposure time is derived, and its applications are studied. Correction for exposure time bias by use of the ratio of scintillation indices with and without time binning is studied. A bandpass-filtered scintillation in a small aperture (computed as the differential-exposure index) provides a reasonably good estimate of the atmospheric time constant for adaptive optics. PMID- 11900149 TI - Sensitivity and information content of aerosol retrievals from the Advanced Very High Resolution Radiometer: radiometric factors. AB - The sensitivity of aerosol optical depths tau1 and tau2 derived from the Advanced Very High Resolution Radiometer (AVHRR) channels 1 and 2, centered at lambda1 = 0.63 and lambda2 = 0.83 microm, respectively, and of an effective Angstrom exponent alpha, derived therefrom as alpha = -ln(tau1/tau2)/ln(lambda1/lambda2), to calibration uncertainties, radiometric noise, and digitization is estimated. Analyses are made both empirically (by introduction of perturbations into the measured radiances and estimation of the respective partial derivatives) and theoretically (by use of a decoupled form of the single-scattering approximation of the radiative transfer equation). The two results are in close agreement. The errors, deltataui and deltaalphai, are parameterized empirically as functions of taui, radiometric errors, and Sun and view geometry. In particular, the alpha errors change in approximately inverse proportion to tau and are comparable with, or even exceed, typical alpha signals over oceans when tau < 0.25. Their detrimental effect on the information content of the AVHRR-derived size parameter gradually weakens as tau increases. PMID- 11900147 TI - Homomorphism between cloudy and clear spectral radiance in the 800-900-cm(-1) atmospheric window region. AB - The sensitivity of a new algorithm for cloud detection over a sea surface has been assessed on the basis of extensive simulations of clear and cloudy radiance spectra, including water and ice and low- and high-altitude clouds. The new algorithm makes use of autocorrelation and cross correlation between an observed spectrum and either a synthetic or a laboratory spectrum and can be used to determine quantitatively the degree of homogeneity of two spectra in the 800-900 cm(-1) region (11.11-12.5 microm). The scheme is intended for high-spectral resolution observations and could form the basis for an operational stand-alone cloud-detection algorithm for next-generation sounding spectrometers. Application of the scheme to real observations is presented and discussed. PMID- 11900150 TI - Ideas at the margin or marginalized ideas? Nonmedical determinants of health in Canada. PMID- 11900148 TI - Ocean-color optical property data derived from the Japanese Ocean Color and Temperature Scanner and the French Polarization and Directionality of the Earth's Reflectances: a comparison study. AB - We describe our efforts to study and compare the ocean-color data derived from the Japanese Ocean Color and Temperature Scanner (OCTS) and the French Polarization and Directionality of the Earth's Reflectances (POLDER). OCTS and POLDER were both on board Japan's Sun-synchronous Advanced Earth Observing Satellite from August 1996 to June 1997, collecting approximately 10 months of global ocean-color data. This operation provided a unique opportunity for the development of methods and strategies for the merging of ocean-color data from multiple ocean-color sensors. We describe our approach to the development of consistent data-processing algorithms for both OCTS and POLDER and the use of a common in situ data set to calibrate vicariously the two sensors. Therefore the OCTS- and POLDER-measured radiances are bridged effectively through common in situ measurements. With this approach to the processing of data from two different sensors, the only differences in the derived products from OCTS and POLDER are the differences that are inherited from the instrument characteristics. Results show that there are no obvious bias differences between the OCTS- and POLDER-derived ocean-color products, whereas the differences due to noise, which stem from variations in sensor characteristics, are difficult to correct at the pixel level. The ocean-color data from OCTS and POLDER therefore can be compared and merged in the sense that there is no significant bias between two. PMID- 11900151 TI - Incorporating socioeconomic factors into U.S. health policy: addressing the barriers. PMID- 11900152 TI - The economics of alcohol abuse and alcohol-control policies. AB - Economic research has contributed to the evaluation of alcohol policy through empirical analysis of the effects of alcohol-control measures on alcohol consumption and its consequences. It has also provided an accounting framework for defining and comparing costs and benefits of alcohol consumption and related policy interventions, including excise taxes. The most important finding from the economics literature is that consumers tend to drink less ethanol, and have fewer alcohol-related problems, when alcoholic beverage prices are increased or alcohol availability is restricted. That set of findings is relevant for policy purposes because alcohol abuse imposes large "external" costs on others. Important challenges remain, including developing a better understanding of the effects of drinking on labor-market productivity. PMID- 11900154 TI - Illegal drug use and public policy. AB - The period from the 1980s to the present has witnessed a lively and unsettled debate concerning the legalization of marijuana, cocaine, heroin, and other illicit substances in the United States. Proponents of legalization argue that the demand for these harmful and potentially addictive substances is not responsive to price. Opponents argue that prices will fall tremendously in a regime characterized by legalization and that the option of legalization and taxation is not feasible. In this paper we summarize theoretical and empirical evidence suggesting that none of these propositions is correct. PMID- 11900155 TI - The economics of tobacco regulation. AB - The past five years have seen a dramatic turn of events against the tobacco industry, raising the question of the appropriate future path for U.S. smoking policy. This paper discusses the theory and evidence on regulation of smoking. I begin by reviewing the background on this industry. I then turn to a discussion of the motivations for regulating smoking, both external and internal to the smoker. I conclude with a discussion of future policy directions. PMID- 11900153 TI - Policy implications of the gradient of health and wealth. AB - Men in the United States with family incomes in the top 5 percent of the distribution in 1980 had about 25 percent longer to live than did those in the bottom 5 percent. Proportional increases in income are associated with equal proportional decreases in mortality throughout the income distribution. I discuss possible reasons for this gradient and ask whether it calls for the redistribution of income in the interest of public health. I argue that the existence of the gradient strengthens the case for income redistribution in favor of the poor but that targeting health inequalities would not be sound policy. PMID- 11900156 TI - Parental behavior and child health. AB - In this paper we document the ways in which parental behavior and socioeconomic status affect children's health. We examine parental behavior in both the prenatal period and childhood. We present evidence on the correlation of this behavior with income and parents' socioeconomic status, and on the ways in which parents' actions affect children's health. We conclude that while health insurance coverage and advances in medical treatment may be important determinants of children's health, they cannot be the only pillars: Protecting children's health also calls for a broader set of policies that target parents' health-related behavior. PMID- 11900157 TI - The environment and health: a conversation with CDC Chief Jeffrey Koplan. Interview by Bernie Goldstein. PMID- 11900158 TI - Whither antitrust? The uncertain future of competition law in health care. AB - Although instrumental in ushering in competition to the health care industry and later in safeguarding the competitive structure of markets, antitrust law has come under attack. A series of questionable judicial decisions has clouded the standards applicable to analyzing health care markets. Legislative efforts to immunize conduct from antitrust challenge also have gathered support in recent years. This study finds scant economic or policy basis for these developments and concludes that anti-managed care sentiments have diluted enthusiasm for applying competitive principles in health care. This phenomenon has resulted in outcome driven judicial decisions and legislative activity geared to serving political expediency rather than sound policy tenets. The paper recommends heightened antitrust scrutiny of provider and insurer markets by federal and state enforcers and increased empirical research into the workings of imperfect health care markets and the effects of past antitrust decisions. PMID- 11900160 TI - Health spending projections for 2001-2011: the latest outlook. AB - This paper describes the most recent ten-year projections of national health spending. projections, produced annually, are based on econometric and actuarial models of the health sector. Our current outlook includes a sharper near-term increase in the health sector's share of gross domestic product (GDP), which reaches 16.8 percent by 2010, compared with the 15.9 percent projected last year. This difference largely reflects legislation-driven increases in public spending growth combined with a weaker economic outlook. Recent acceleration in private sector health spending is projected to peak in 2002. PMID- 11900161 TI - The financial performance of community health centers, 1996-1999. AB - This paper presents recent financial health trends of community health centers (CHCs) between 1996 and 1999, a time characterized by fiscal and operating challenges. Results show that many individual CHCs have been subject to large changes in payer-mix among uninsured and Medicaid users. Troubling is the finding that more than half of all CHCs reported operating deficits in 1997, 1998 and 1999. CHCs experiencing large increases in the share of uninsured users and those participating in Medicaid managed care appear to have been disproportionately affected. The analyses presented support recommendations for enhanced data collection and for further monitoring of CHCs' financial health. PMID- 11900159 TI - How disclosing HMO physician incentives affects trust. AB - Opinions are deeply divided over whether rewarding physicians for lowering costs decreases trust in physicians or insurers. To explore the effects of disclosing physician payment methods in HMOs, members of two similar HMO plans were randomized to intervention and control groups, and the experimental arm was told how the HMO paid their primary care physician. Separate disclosures were developed for each plan, one describing primarily capitation payment, and the other (mixed-incentive plan) describing fee-for-service payment with a bonus that rewards cost savings, satisfaction, and preventive services. The disclosures pointed out more of the positive than the negative features of these incentives. We found that the disclosures doubled the number of subjects with substantial knowledge of the physician incentives and halved the number with no knowledge. Nevertheless, the disclosures had no negative effects on trust of either physicians or insurers. The capitated plan disclosure had a small positive effect on trust of physicians. Disclosing the positive and negative features of incentives and increasing knowledge of these incentives does not, in the short term, reduce trust in physicians or insurers and may have a mild positive impact on physician trust, perhaps as a consequence of displaying candor and increasing understanding of positive features. PMID- 11900162 TI - No free lunch. PMID- 11900163 TI - A matter of influence. PMID- 11900164 TI - Between the lines: navigating the uncharted territory of industry-sponsored research. PMID- 11900165 TI - The benefits and risks of a pack of M&Ms. PMID- 11900167 TI - Youth targeting by tobacco manufacturers since the Master Settlement Agreement. AB - The 1998 Master Settlement Agreement (MSA) between tobacco manufacturers and forty-six states bans manufacturers from targeting minors through advertising. To determine how youth targeting in magazine cigarette advertisements changed after the MSA, we analyzed magazine readership and cigarette ads in U.S. magazines from 1997 to 2000. In 2000 all three major manufacturers (Philip Morris, R.J. Reynolds, and Brown and Williamson) failed to comply with the MSA's youth targeting ban, selectively increasing their youth targeting. Banning all magazine advertising of cigarettes may be necessary to eliminate youth targeting in magazines. PMID- 11900166 TI - The effects of obesity, smoking, and drinking on medical problems and costs. AB - This paper compares the effects of obesity, overweight, smoking, and problem drinking on health care use and health status based on national survey data. Obesity has roughly the same association with chronic health conditions as does twenty years' aging; this greatly exceeds the associations of smoking or problem drinking. Utilization effects mirrors the health effects. Obesity is associated with a 36 percent increase in inpatient and outpatient spending and a 77 percent increase in medications, compared with a 21 percent increase in inpatient and outpatient spending and a 28 percent increase in medications for current smokers and smaller effects for problem drinkers. Nevertheless, the latter two groups have received more consistent attention in recent decades in clinical practice and public health policy. PMID- 11900168 TI - Power of information: closing the gap between research and policy. AB - States play an increasing role in setting U.S. health policy. A survey of 292 state government policymakers finds that officials are overwhelmed by the volume of information they receive and have a strong preference for information that is concise and more relevant to current debates. Younger officials are more likely to use electronic information, while older policymakers prefer printed material. Organizations of government professionals are trusted sources of information, and state agencies are a key source of data and information. Policymakers expressed a strong desire for tools to help them identify research on specific topics. PMID- 11900169 TI - The California Wellness Foundation's grant making in work and health. PMID- 11900170 TI - The coming physician shortage. PMID- 11900171 TI - The hospital crisis is here. PMID- 11900172 TI - California's hospitals. PMID- 11900173 TI - New York hospital leaders bear the blame. PMID- 11900174 TI - Hospital deregulation in New York. PMID- 11900175 TI - New York working through its problems. PMID- 11900176 TI - New York legislature got it right. PMID- 11900177 TI - Top-down nursing changes needed. PMID- 11900178 TI - Nurses devalued and abandoned. PMID- 11900179 TI - Improving nursing conditions. PMID- 11900180 TI - Value purchasing and quality. PMID- 11900181 TI - Public reporting on nursing homes. PMID- 11900182 TI - Acute illness pilot projects needed. PMID- 11900183 TI - Improving chronic care. PMID- 11900184 TI - Autologous bone marrow transplants. PMID- 11900185 TI - The influence of income on health: views of an epidemiologist. AB - Income is related to health in three ways: through the gross national product of countries, the income of individuals, and the income inequalities among rich nations and among geographic areas. A central question is the degree to which these associations reflect a causal association. If so, redistribution of income would improve health. This paper discusses two ways in which income could be causally related to health: through a direct effect on the material conditions necessary for biological survival, and through an effect on social participation and opportunity to control life circumstances. The fewer goods and services are provided publicly by the community, the more important individual income is for health. Under present U.S. circumstances, a policy of counteracting growing income inequalities through the tax and benefit system and of public provision appears justified. PMID- 11900186 TI - Disadvantage, inequality, and social policy. AB - Eliminating disparities in health is a primary goal of the federal government and many states. Our overarching objective should be to improve population health for all groups to the maximum extent. Ironically, enhancing population health and even the health of the disadvantaged can conflict with efforts to reduce disparities. This paper presents data showing that interventions that offer some of the largest possible gains for the disadvantaged may also increase disparities, and it examines policies that offer the potential to decrease disparities while improving population health. Enhancement of educational attainment and access to health services and income support for those in greatest need appear to be particularly important pathways to improved population health. PMID- 11900187 TI - Socioeconomic disparities in health: pathways and policies. AB - Socioeconomic status (SES) underlies three major determinants of health: health care, environmental exposure, and health behavior. In addition, chronic stress associated with lower SES may also increase morbidity and mortality. Reducing SES disparities in health will require policy initiatives addressing the components of socioeconomic status (income, education, and occupation) as well as the pathways by which these affect health. Lessons for U.S. policy approaches are taken from the Acheson Commission in England, which was charged with reducing health disparities in that country. PMID- 11900188 TI - The case for more active policy attention to health promotion. AB - Until recently, when anthrax triggered a concern about preparedness in the public health infrastructure, U.S. health policy and health spending had been dominated by a focus on payment for medical treatment. The fact that many of the conditions driving the need for treatment are preventable ought to draw attention to policy opportunities for promoting health. Following a brief review of the determinants of population health-genetic predispositions, social circumstances, environmental conditions, behavioral patterns, and medical care-this paper explores some of the factors inhibiting policy attention and resource commitment to the nonmedical determinants of population health and suggests approaches for sharpening the public policy focus to encourage disease prevention and health promotion. PMID- 11900189 TI - What the federal government can do about the nonmedical determinants of health. AB - Growing recognition that the acute health care delivery system contributes proportionally less to health when compared with environment and behavior has focused scholars and public health experts on the need to address nonmedical determinants of health. This paper outlines some steps that the U.S. government can take to address these factors and describes some of the challenges involved. Actions that can be undertaken now are increased education and leadership, development of mechanisms to further collaboration among sectors, expanded monitoring and reporting on nonmedical determinants, and developing new knowledge about how these factors affect health and successful interventions to address them. PMID- 11900190 TI - On MEG forward modelling using multipolar expansions. AB - Magnetoencephalography (MEG) is a non-invasive functional imaging modality based on the measurement of the external magnetic field produced by neural current sources within the brain. The reconstruction of the underlying sources is a severely ill-posed inverse problem typically tackled using either low-dimensional parametric source models, such as an equivalent current dipole (ECD), or high dimensional minimum-norm imaging techniques. The inability of the ECD to properly represent non-focal sources and the over-smoothed solutions obtained by minimum norm methods underline the need for an alternative approach. Multipole expansion methods have the advantages of the parametric approach while at the same time adequately describing sources with significant spatial extent and arbitrary activation patterns. In this paper we first present a comparative review of spherical harmonic and Cartesian multipole expansion methods that can be used in MEG. The equations are given for the general case of arbitrary conductors and realistic sensor configurations and also for the special cases of spherically symmetric conductors and radially oriented sensors. We then report the results of computer simulations used to investigate the ability of a first-order multipole model (dipole and quadrupole) to represent spatially extended sources, which are simulated by 2D and 3D clusters of elemental dipoles. The overall field of a cluster is analysed using singular value decomposition and compared to the unit fields of a multipole, centred in the middle of the cluster, using subspace correlation metrics. Our results demonstrate the superior utility of the multipolar source model over ECD models in providing source representations of extended regions of activity. PMID- 11900191 TI - Electric fields induced in a spherical volume conductor by temporally varying magnetic field gradients. AB - A homogeneous spherical volume conductor is used as a model system for the purpose of calculating electric fields induced in the human head by externally applied time-varying magnetic fields. We present results for the case where magnetic field gradient coils, used in magnetic resonance imaging (MRI), form the magnetic field, and we use these data to put limits on the rates of gradient change with time needed to produce nerve stimulation. The electric field is calculated analytically for the case of ideal longitudinal and transverse linear field gradients. We also show results from computer calculations yielding the electric field maps in a sphere when the field gradients are generated by a real MRI gradient coil set. In addition, the effect of shifting the sphere within each gradient coil volume is investigated. Numerical analysis shows similar results when applied to a model human head. PMID- 11900192 TI - Small-angle x-ray scattering studies of human breast tissue samples. AB - Small-angle x-ray scattering (SAXS) patterns are recorded from thin breast tissue samples containing healthy and cancerous regions. The SAXS patterns are compared with histo-pathological observations. The information available from SAXS is reviewed, and a model for scattering from collagen is presented. Scattering patterns of collagen at regions far from the tumours are essentially different from those at tumours. The axial period of collagen fibrils is 65.0 +/- 0.1 nm in healthy regions, and 0.3 nm larger in cancer-invaded regions. The average intensity of scattering from cancerous regions is an order of magnitude higher than the intensity from healthy regions. This is interpreted to arise from an increase of the specific surface area of the scatterers, which is due to a disruption of the molecular and supra-molecular structures in cancerous regions and invasion of new types of cells. The differences of the SAXS patterns are large and distinctive enough to suggest that these phenomena may be utilized in mammography. PMID- 11900193 TI - Precise modelling of the eye for proton therapy of intra-ocular tumours. AB - A new method is described that allows precise modelling of organs at risk and target volume for radiation therapy of intra-ocular tumours. The aim is to optimize the dose distribution and thus to reduce normal tissue complication probability. A geometrical 3D model based on elliptic shapes was developed that can be used for multimodal model-based segmentation of 3D patient data. The tumour volume cannot be clearly identified in CT and MR data, whereas the tumour outline can be discriminated very precisely in fundus photographs. Therefore, a multimodal 2D fundus diagram was developed, which allows us to correlate and display simultaneously information extracted from the eye model, 3D data and the fundus photograph. Thus, the connection of fundus diagram and 3D data is well defined and the 3D volume can be calculated directly from the tumour outline drawn onto the fundus photograph and the tumour height measured by ultrasound. The method allows the calculation of a precise 3D eye model of the patient, including the different structures of the eye as well as the tumour volume. The method was developed as part of the new 3D treatment planning system OCTOPUS for proton therapy of ocular tumours within a national research project together with the Hahn-Meitner-Institut Berlin. PMID- 11900195 TI - Performance analysis and determination of the p(wall) correction factor for 60Co gamma-ray beams for Wellhofer Roos-type plane-parallel chambers. AB - The wall perturbation correction factor p(wall) in 60Co for Wellhofer Roos-type plane-parallel ionization chambers is determined experimentally and compared with the results of a previous study using PTW-Roos chambers (Palm et al 2000 Phys. Med. Biol. 45 971-81). Five ionization chambers of the type Wellhofer PPC-35 (or its equivalent PPC-40) are used for the analysis. Wall perturbation correction factors are obtained by assuming N(D,air) chamber factors determined by cross calibration in a high-energy electron and in a 60Co gamma-ray beam to be equal, and by assigning any differences to the wall perturbation factor. The procedure yields a p(wall) value of 1.018 (u(c) = 0.010), which is slightly higher than the value 1.014 (u(c) = 0.010) formerly obtained for the PTW-Roos chambers using the N(D,air) method. The chamber-to-chamber variation in p(wall) for the Wellhofer Roos chambers is found to be very small, with a maximum difference of 0.3%. The effect of using new p(cav) values for graphite-walled Farmer-type chambers used in water in electron beams is to decrease p(wall) by approximately 0.5%. The long and short-term stability of the Roos-type chambers manufactured by Wellhofer is investigated by measurements at the IAEA Dosimetry Laboratory in Vienna, Austria, and at the Sahlgrenska University Hospital in Goteborg, Sweden. Calibrations made at the IAEA over several months show variations in the N(D,w) calibration factors larger than expected. based on previous experiences with PTW-Roos chambers. Measurements of the short-term stability of the Wellhofer-Roos chambers show a marked increase in chamber response for the time the chambers are immersed in water, pointing to a possible problem in the chamber design. As a consequence of these findings, Wellhofer is currently working on a re-design of the chamber to solve the stability problem. PMID- 11900194 TI - Production, PET performance and dosimetric considerations of 134Ce/134La, an Auger electron and positron-emitting generator for radionuclide therapy. AB - We propose the use of the Auger electron and positron-emitting generator 134Ce/134La (half-lives 3.16 d and 6.45 min) for radionuclide therapy. It combines emission of high-energy beta particles with Auger electrons. The high energy beta particles have similar energies as those emitted by 90Y. Many cancer patients receiving radionuclide therapy have both bulk tumours, which are best treated with high-energy beta particles, and single spread cells or micrometastasis, which are preferably treated with low-energy electrons such as Auger and conversion electrons. Furthermore, the positron-emitting 134La can be used to study kinetics and dosimetry using PET. Production and PET performance were investigated and theoretical dosimetry calculations were made. PET resolution, recovery and quantitative accuracy were slightly degraded for 134La compared to 18F. 134Ce/134La absorbed doses to single cells were higher than absorbed doses from 90Y and 111In. Absorbed doses to spheres representing bulk tumours were almost as high as for 90Y, and a factor 10 higher than for 111In. Whole-body absorbed doses, based on kinetics of the somatostatin analogue octreotide, were higher for 134Ce/134La than for 90Y because of the 134La annihilation photons. This initial study of the therapeutic possibilities of 134Ce/134La is encouraging and justifies further investigations. PMID- 11900196 TI - Image processing techniques for noise removal, enhancement and segmentation of cartilage OCT images. AB - Osteoarthritis, whose hallmark is the progressive loss of joint cartilage, is a major cause of morbidity worldwide. Recently, optical coherence tomography (OCT) has demonstrated considerable promise for the assessment of articular cartilage. Among the most important parameters to be assessed is cartilage width. However, detection of the bone cartilage interface is critical for the assessment of cartilage width. At present, the quantitative evaluations of cartilage thickness are being done using manual tracing of cartilage-bone borders. Since data is being obtained near video rate with OCT, automated identification of the bone cartilage interface is critical. In order to automate the process of boundary detection on OCT images, there is a need for developing new image processing techniques. In this paper we describe the image processing techniques for speckle removal, image enhancement and segmentation of cartilage OCT images. In particular, this paper focuses on rabbit cartilage since this is an important animal model for testing both chondroprotective agents and cartilage repair techniques. In this study, a variety of techniques were examined. Ultimately, by combining an adaptive filtering technique with edge detection (vertical gradient, Sobel edge detection), cartilage edges can be detected. The procedure requires several steps and can be automated. Once the cartilage edges are outlined, the cartilage thickness can be measured. PMID- 11900197 TI - Coherent thermal wave imaging of subsurface chromophores in biological materials. AB - Thermal wave imaging of discrete subsurface chromophores in biological materials is reported using a phase sensitive coherent detection technique applied to recorded infrared (IR) images. We demonstrate that utilization of a periodically modulated laser source for thermal wave excitation and coherent detection applied to each pixel may be used to compute images of thermal wave amplitude and phase at the laser modulation frequency. In comparison to recorded IR images, the narrow-band detection technique significantly improves the quality of thermal wave amplitude images of subsurface chromophores in biological materials. Additionally, the technique provides phase information, which may be used to estimate chromophore depth in tissue. Application of the technique is demonstrated using tissue phantoms and in vivo biological models. We present a theoretical analysis and computer simulations that demonstrate the effect of tissue optical and thermal properties on thermal wave amplitude and phase. In comparison to the pulsed photothermal technique, coherent thermal wave imaging of subsurface chromophores in tissue for diagnostic applications allows reduction of peak incident laser fluence by as much as four orders of magnitude and is safer and more amenable to in vivo imaging. PMID- 11900198 TI - Measurements of lead in human tibiae. A comparison between K-shell x-ray fluorescence and electrothermal atomic absorption spectrometry. AB - The aim of this study was to validate 109Cd-based K-shell x-ray fluorescence measurements against atomic absorption spectrometry (AAS) measurements of core and surface tibia lead. The lead content of nine adult human cadaver tibiae was measured using 109Cd-based K-shell x-ray fluorescence (XRF) spectrometry and the results compared to measurements obtained using electrothermal atomization atomic absorption spectrometry following acid digestion. Each tibia was divided into nine cross-sectional segments, which were further separated into tibia core and surface samples for the AAS analytical measurements. Proximal-distal variability in tibia lead concentration as determined by AAS was previously described for both surface and core segments and was found to decrease towards the ends of the tibia, in contrast to XRF in which lead was found to increase towards the tibia ends. The effect of this contrasting behaviour on the agreement between XRF and AAS measurements was examined. Lead concentrations determined by AAS ranged from 3 to 19 microg of lead per gram of dry weight bone (microg g(-1)) for tibia core and from 5 to 32 microg g(-1) for tibia surface. Lead concentrations determined by XRF ranged from 2 to 35 microg g(-1) dry weight. No statistically significant difference was found between mean XRF-measured concentrations and mean surface lead concentrations measured by AAS, but XRF significantly overestimated tibia core lead concentrations by between 5 and 8 microg g(-1). PMID- 11900199 TI - A quantitative evaluation of spurious results in the infrared spectroscopic measurement of CO2 isotope ratios. AB - The possible generation of spurious results, arising from the application of infrared spectroscopic techniques to the measurement of carbon isotope ratios in breath, due to coincident absorption bands has been re-examined. An earlier investigation, which approached the problem qualitatively, fulfilled its aspirations in providing an unambiguous assurance that 13C16O2/12C16O2 ratios can be confidently measured for isotopic breath tests using instruments based on infrared absorption. Although this conclusion still stands, subsequent quantitative investigation has revealed an important exception that necessitates a strict adherence to sample collection protocol. The results show that concentrations and decay rates of the coincident breath trace compounds acetonitrile and carbon monoxide, found in the breath sample of a heavy smoker, can produce spurious results. Hence, findings from this investigation justify the concern that breath trace compounds present a risk to the accurate measurement of carbon isotope ratios in breath when using broadband, non-dispersive, ground state absorption infrared spectroscopy. It provides recommendations on the length of smoking abstention required to avoid generation of spurious results and also reaffirms, through quantitative argument, the validity of using infrared absorption spectroscopy to measure CO2 isotope ratios in breath. PMID- 11900200 TI - Intravascular brachytherapy of the coronary arteries. AB - This is a review of the relatively recently developed field of intravascular brachytherapy of coronary arteries. It presents a brief overview of the discipline of coronary angioplasty describing the problem of restenosis and discusses the potential for ionizing radiation to overcome this problem. It examines the various methods that have been used to irradiate the coronary arteries comparing their advantages and disadvantages. Special consideration is given to seeds and wires in the artery, radioactive liquids in the angioplasty balloon and radioactive stents. Passing reference is made to a number of other methods that have also been proposed, but which are not commonly used to irradiate the coronary arteries at present. The dosimetry of each of the major techniques is discussed and the data from different laboratories compared. Specific consideration is given to the need for centring of the radioactive source and the factors affecting the selection of a dose prescription. A brief review of recent clinical trials is followed by an examination of possible future directions in this field including the use of intravascular ultrasound to improve dosimetry, the use of gas-filled balloons to enhance the penetration of beta emitting sources and the use of gamma-emitting stents to overcome the problems associated with edge restenosis. PMID- 11900201 TI - Plutonium and americium inventories in atmospheric fallout and sediment cores from Blelham Tarn, Cumbria (UK). AB - The objective of this paper is to report on the results of a study of 238Pu, 239 + 240Pu and 241Am inventories onto Blelham Tarn in Cumbria (UK). The atmospheric fallout inventory was obtained by analysing soil cores and the results are in good agreement with the literature: 101 Bq m(-2) for 239 + 240Pu; 4.5 Bq m(-2) for 238Pu and 37 Bq m(-2) for 241Am. The sediment core inventory for the whole lake is compared to the atmospheric fallout inventory. The sediment activity is 60-80% higher than the estimated fallout activity, showing a catchment area contribution and in particular the stream input. PMID- 11900203 TI - A general methodology for structuring models to predict the long-term migration of radionuclides from catchments. AB - In the present paper a generic model for predicting the long-term migration of radionuclides and heavy metals from catchments is described. The model subdivides the catchment into a number of homogeneous, infinitesimal sub-catchments and integrates the radionuclide contributions from such sub-catchments to calculate the total flux of contaminant. It relates the radionuclide behaviour to the statistical distribution of the pollutant partition coefficient on the "ensemble" of sub-catchments. The methodology was validated for 90Sr and 137Cs by using data for water contamination in some European rivers. Values of migration parameters for Pu, Tc, I and Cd isotopes were obtained. PMID- 11900202 TI - Detection and analysis of xenon isotopes for the comprehensive nuclear-test-ban treaty international monitoring system. AB - The use of the xenon isotopes for detection of nuclear explosions is of great interest for monitoring compliance with the comprehensive nuclear-test-ban treaty (CTBT). Recently, the automated radioxenon sampler-analyzer (ARSA) was tested at the Institute for Atmospheric Radioactivity (IAR) in Freiburg, Germany to ascertain its use for the CTBT by comparing its results to laboratory-based analyses, determining its detection sensitivity and analyzing its results in light of historical xenon isotope levels and known reactor operations in the area. Xe-133 was detected nearly every day throughout the test at activity concentrations ranging between approximately 0.1 mBq/m3 to as high as 120 mBq/m3. Xe-133m and 135Xe were also detected occasionally during the test at concentrations of less than 1 to a few mBq/m3. PMID- 11900204 TI - Ecological half-life of 137Cs in plants associated with a contaminated stream. AB - Ecological half-life (Te) is a useful measure for studying the long-term decline of contaminants, such as radionuclides, in natural systems. The current investigation determined levels of radiocesium (137Cs) in two aquatic (Polygonum punctatum, Sagittaria latifolia) and three terrestrial (Alnus serrulata, Myrica cerifera, Salix nigra) plant species from a contaminated stream and floodplain on the U.S. Department of Energy's Savannah River Site. Current 137Cs levels in plants were used in conjunction with historical data to determine Te of 137Cs in each species. Median concentrations of 137Cs were highest in S. latifolia (0.84 Bq g(-1)) and lowest in M. cerifera (0.10 Bq g(-1)). Te's ranged from 4.85 yr in M. cerifera to 8.35 yr in S. nigra, both terrestrial species. Te's for all aquatic (6.30 yr) and all terrestrial (5.87) species combined were very similar. The Te's of the two aquatic primary producers (P. punctatum and S. latifolia) in the Steel Creek ecosystem were somewhat longer than Te values previously reported for some consumers from this ecosystem. PMID- 11900205 TI - Sample preparation for the determination of 241Am in sediments utilizing gamma spectroscopy. AB - This paper describes a procedure developed to separate americium-241 from the bulk of a sample by coprecipitation followed by high sensitivity gamma-counting of the concentrate in a well-type detector. It enables the measurement of 241Am at low concentrations, e.g. fallout levels in soils and sediments, or where large sample sizes are not available. The method is much faster and more reliable than those involving separation from other alpha-emitters, electroplating and alpha spectrometry. A number of tracer experiments was performed in order to optimize the conditions for coprecipitation of 241Am from sediment leachates. The general outline of the determination of americium is also given. PMID- 11900206 TI - Significant radioactive contamination of soil around a coal-fired thermal power plant. AB - Soil samples were collected around a coal-fired power plant from 81 different locations. Brown coal, unusually rich in uranium, is burnt in this plant that lies inside the confines of a small industrial town and has been operational since 1943. Activity concentrations of the radionuclides 238U, 226Ra, 232Th, 137Cs and 40K were determined in the samples. Considerably elevated concentrations of 238U and 226Ra have been found in most samples collected within the inhabited area. Concentrations of 235U and 226Ra in soil decreased regularly with increasing depth at many locations, which can be explained by fly-ash fallout. Concentrations of 235U and 226Ra in the top (0-5 cm depth) layer of soil in public areas inside the town are 4.7 times higher, on average, than those in the uncontaminated deeper layers, which means there is about 108 Bq kg(-1) surplus activity concentration above the geological background. A high emanation rate of 222Rn from the contaminated soil layers and significant disequilibrium between 238U and 226Ra activities in some kinds of samples have been found. PMID- 11900207 TI - Reconstructing the history of 14C discharges from Sellafield: part 1--atmospheric discharges. AB - 14C specific activities, above ambient background levels, were determined in individual treerings (corresponding to the years 1950-1999) sectioned from an oak tree that was felled in autumn 1999, from a location 1.5 km east of the Sellafield nuclear fuel reprocessing plant in Cumbria. north-west England. The data were used to produce a new, improved, reconstruction of Sellafield's annual atmospheric 14C discharges between 1951 and 1999, using the most reliable discharge data set (1994-1999) as the primary basis for the determination of a new calibration factor that relates excess 14C activity in individual tree rings to the annual discharge during the corresponding year. The results indicate that the current British Nuclear Fuels plc (BNFL) estimate of total 14C discharges to the atmosphere prior to 1978 is significantly overestimated, while the current estimate of total 14C discharges after 1978 is very similar to that determined in this study. In this study, the total activity of 14C discharged to the atmosphere from Sellafield between 1951 and 1999 is estimated to be 259+/-63 TBq (at 2 std. dev.). The BNFL current estimate is 360TBq. PMID- 11900208 TI - Uptake of 40K and 137Cs in native plants of the Marshall Islands. AB - Uptake of 137Cs and 40K was studied in seven native plant species of the Marshall Islands. Plant and soil samples were obtained across a broad range of soil 137Cs concentrations (0.08-3900 Bq/kg) and a narrower range of 40K soil concentrations (2.3-55 Bq/kg), but with no systematic variation of 40K relative to 137Cs. Potassium-40 concentrations in plants varied little within the range of 40K soil concentrations observed. Unlike the case for 40K, 137Cs concentrations increased in plants with increasing 137Cs soil concentrations though not precisely in a proportionate manner. The best-fit relationship between soil and plant concentrations was P = aSb where a and b are regression coefficients and P and S are plant and soil concentrations, respectively. The exponent b for 40K was zero, implying plant concentrations were a single value, while b for 137Cs varied between 0.51 and 0.82, depending on the species. For both 40K and 137Cs, we observed a decreasing concentration ratio (where concentration ratio=plant concentration/soil concentration) with increasing soil concentrations. For the CR values, the best-fit relationship was of the form CR = aSb/S = aSb(-1). For the 40K CR functions, the exponent b - 1 was close to - 1 for all species. For the 137Cs CR functions, the exponent b - 1 varied from -0.19 to -0.48. The findings presented here, aswell as those by other investigators, collectively argue against the usefulness of simplistic ratio models to accurately predict uptake of either 40K or 137Cs in plants over wide ranges of soil concentration. PMID- 11900209 TI - Goserelin (Zoladex)--its role in early breast cancer in pre- and perimenopausal women. AB - Current standard adjuvant therapies for premenopausal women with early breast cancer include ovarian ablation by surgery or irradiation, chemotherapy and tamoxifen. The value of ovarian ablation in prolonging the survival of premenopausal patients with early breast cancer was clearly established by the analyses performed by the Early Breast Cancer Trialists' Collaborative Group in 1996. More recently, the value of ovarian suppression using the luteinizing hormone releasing hormone analogue goserelin as adjuvant therapy in pre /perimenopausal women with early breast cancer has been confirmed in a series of studies involving over 8000 patients. The results from these studies provide evidence that goserelin, alone or in combination with tamoxifen, is at least as effective as cytotoxic chemotherapy in patients with hormone receptor-positive tumours and is effective when used after adjuvant chemotherapy. The use of goserelin in the management of early breast cancer presents an option which can avoid the side-effects experienced with cytotoxic chemotherapy and may offer unique benefits to premenopausal patients. The consolidation of these emerging results should help in defining the optimal role for goserelin in pre /perimenopausal patients with early breast cancer. PMID- 11900210 TI - ICI 182,780 (Fulvestrant)--the first oestrogen receptor down-regulator--current clinical data. AB - ICI 182,780 (Fulvestrant) is the first in a new class of novel, steroidal, 'pure' antioestrogens--the oestrogen receptor (ER) down-regulators. Its unique mode of action and the absence of partial agonist activity make it a candidate for the treatment of advanced breast cancer in both pre- and postmenopausal women. Tamoxifen has been available for use over the past 25 years. However, its partial agonist activity has been associated with detrimental effects, particularly on the endometrium, and may be associated with the development of tamoxifen resistance. Other antioestrogen agents have previously been unable to demonstrate clinically relevant activity following the development of resistance to tamoxifen. In contrast, the unique mechanism of action of ICI 182,780 results in significant clinical activity in patients failing on tamoxifen therapy. Indeed, phase III clinical trials have demonstrated that ICI 182,780 is at least as effective as the aromatase inhibitor anastrozole in the treatment of postmenopausal patients with advanced disease who have progressed during threatment with prior enocrine therapy. As such, ICI 182,780 will provide a valuable addition to the armamentarium for the treatment of advanced breast cancer. PMID- 11900211 TI - A vision for the future? AB - Systemic adjuvant therapy is recommended immediately following surgical removal of the primary tumour in the majority of patients with early breast cancer, to prevent the recurrence of distant metastases. Significant progress has been made in the development and evaluation of endocrine therapies for systemic adjuvant therapy. In pre- and perimenopausal women, ovarian ablation has proven to be a valuable treatment option, though not always desirable for young patients. Thus, reversible medical ovarian suppression with a luteinizing hormone releasing hormone agonist, such as goserelin (Zoladex), may provide an attractive alternative for such patients. International trials have indicated that goserelin provides an important addition to the choice of adjuvant therapies now available to pre- and perimenopausal patients. For postmenopausal patients, it is hoped that the ATAC (Arimidex, tamoxifen, alone or in combination) trial will reveal whether or not the benefits of anastrozole (Arimidex) observed in advanced disease, where it has proven to be well tolerated and at least as effective as tamoxifen in recent trials, will translate to the early setting to provide further management options for these patients. On the horizon is yet another exciting endocrine agent, ICI 182,780 (Fulvestrant), which has also been shown to be as effective as anastrozole in advanced disease. In terms of the future, these agents are likely to provide additional valuable treatment choices for early breast cancer across the patient spectrum. PMID- 11900212 TI - Anastrozole (Arimidex)--an aromatase inhibitor for the adjuvant setting? AB - Anastrozole (Arimidex) is a third-generation aromatase inhibitor which has been shown to possess superior efficacy and tolerability over established endocrine agents in advanced breast cancer. Inhibition of aromatase prevents the conversion of androgen substrates to oestrogen, its sole source in postmenopausal women, thereby leading to regression of hormone-sensitive breast carcinomas. Clinical pharmacology data indicate that anastrozole is a potent aromatase inhibitor, providing near-maximal suppression of serum and intratumoural oestrogens to below detectable levels. Anastrozole may offer greater selectivity compared with other aromatase inhibitors, being without any intrinsic endocrine effects and with no apparent effect on the synthesis of adrenal steroids. It is well tolerated and has a convenient once-daily dosing regimen, ensuring maximum patient compliance. A major clinical programme has demonstrated that anastrozole is superior to the standard endocrine therapy, tamoxifen, for the first-line treatment of postmenopausal women with hormone-sensitive advanced breast cancer. Its superior efficacy in advanced disease, together with its improved tolerability and convenient dosage, make it a suitable agent to be assessed for the treatment of early breast cancer in postmenopausal women. This was investigated in the largest single adjuvant breast cancer study ever to be carried out, the ATAC (Arimidex, tamoxifen, alone or in combination) trial, which has now completed recruitment, with the first efficacy and safety data awaited. PMID- 11900213 TI - Comparison of two different schedules of granulocyte-colony-stimulating factor during treatment for acute lymphocytic leukemia with a hyper-CVAD (cyclophosphamide, doxorubicin, vincristine, and dexamethasone) regimen. AB - BACKGROUND: Although the safety and efficacy of granulocyte-colony-stimulating factor (G-CSF) (filgrastim) in the treatment of hematologic malignancies has been well established, to the authors' knowledge the optimal timing of filgrastim administration during remission induction chemotherapy and consolidation chemotherapy has not been determined. The purpose of the current study was to determine whether a delay in the administration of filgrastim from Day 5 to Day 10 during chemotherapy with a hyper-CVAD (cyclophosphamide, doxorubicin, vincristine, and dexamethasone) regimen resulted in a longer time to neutrophil or platelet count recovery or increased the incidence of infection. METHODS: One hundred ninety-nine patients who achieved complete disease remission after a single course of induction chemotherapy were considered for evaluation. Induction chemotherapy was with hyper-CVAD (fractionated cyclophosphamide, 300 mg/m2, twice daily for Days 1-3; doxorubicin, 50 g/m2, on Day 4; vincristine, 2 mg, on Days 4 and 11; and dexamethasone, 40 mg, on Days 1-4 and Days 11-14), which also was given in odd-numbered consolidation Courses 3, 5, and 7. Even-numbered courses (Courses 2, 4, 6, and 8) were comprised of methotrexate, 200 mg/m2, over 2 hours followed by 800 mg/m2 over 24 hours on Day 1; cytarabine, 3 g/m2, every 12 hours for 4 doses over 2 days (Days 2 and 3); and intravenous methylprednisolone, 50 mg, twice daily on Days 1-3 (MTX/ara-C regimen). Two sequential treatment groups were assessable based on timing of the filgrastim administration; 151 patients received filgrastim starting on Day 5 (D5) of induction chemotherapy and 48 patients received filgrastim starting on Day 10 (D10). RESULTS: Time to neutrophil recovery was shorter for the D5 group than for the D10 group during induction chemotherapy (18 days vs. 19 days; P = 0.04) and hyper-CVAD Courses 3 and 5 (12 days vs. 15 days during Course 3, P < 0.001; and 13 days vs. 16 days during Course 5, P = 0.002). There was no apparent significant difference between the two groups with regard to time to neutrophil recovery during the MTX/ara-C courses or the last hyper-CVAD course. Delay in the administration of filgrastim did not appear to result in an increase in time to platelet count recovery or in the incidence of infection; however, there was an increased incidence of mucositis during induction chemotherapy. CONCLUSIONS: For a hyper-CVAD and MTX/ara-C regimen, the results of the current study have shown that the administration of filgrastim can be delayed until Day 10 without increasing the risk of treatment-related morbidity during consolidation chemotherapy. During induction chemotherapy, delay in the administration of filgrastim may result in a slight increase in the time to neutrophil count recovery and risk of mucositis, but there is no apparent associated increase in the risk of infection. PMID- 11900214 TI - Ductal lavage and the clinical management of women at high risk for breast carcinoma: a commentary. PMID- 11900215 TI - A randomized, multicenter prospective trial assessing long-acting release octreotide pamoate plus tamoxifen as a first line therapy for advanced breast carcinoma. AB - BACKGROUND: Long-acting release octreotide pamoate (OP-LAR) is a synthetic octapeptide that can be administered monthly and whose activity is similar to that of endogenous somatostatin. In vitro and in vivo data suggest that OP-LAR may act as a growth inhibitor or a modulator of growth stimulatory peptides. The potential mechanisms of action of somatostatin analogues in breast carcinoma include the suppression of insulin-like growth factor-1 (a putative tumor growth factor) and the binding to the somatostatin receptors expressed by breast carcinoma cells in order to induce apoptosis. METHODS: This Phase III, multicenter, randomized, double-blind, placebo-controlled trial involved 203 patients (13% premenopausal and 87% postmenopausal) with locally recurrent (but unsuitable for local treatment) or metastatic breast carcinoma, 199 of whom were actually treated (99 patients with OP-LAR and 100 patients with placebo). All patients received TAM and were estrogen and/or progesteron receptor positive (receptor positivity was an eligibility criterion), and all had measurable or evaluable disease. Any patients who had received previous chemotherapy not given in an adjuvant or neoadjuvant setting were excluded. RESULTS: At the time of the interim analysis, the tumor response rates (RR) were 20.2% (9 complete responses [CR] and 11 partial responses [PR]) in the OP-LAR arm and 21% (11 CRs and 10 PRs) in the placebo arm, and the median time to progression (TTP) was 25.0 and 26.9 weeks (P = 0.62), respectively. The adverse events experienced by 10% or more of the patients and attributed to octreotide were gastrointestinal in nature: diarrhea (53%), nausea (16%), and abdominal pain (11%). CONCLUSIONS: Because of the similar RR and TTP in both arms, the trial was stopped at the interim analysis. The current data confirm there is no indication for adding somatostatin analogues to TAM in advanced breast carcinoma. PMID- 11900216 TI - Model-based predictions of BRCA1/2 mutation status in breast carcinoma patients treated at an academic medical center. AB - BACKGROUND: Women with an existing breast carcinoma diagnosis who are found to carry a BRCA1/2 mutation have a substantial risk of developing both a contralateral breast carcinoma and ovarian carcinoma. In a newly diagnosed breast carcinoma patient, this genetic information may influence the management of her disease. To assess the volume of patients who may need genetic services at the time of diagnosis, the authors determined the proportion of women with newly diagnosed breast carcinoma at the study institution who would be eligible for genetic testing. METHODS: Fifty consecutive women with new breast carcinoma who were attending a multidisciplinary clinic were interviewed. Detailed, three generation pedigrees were collected for each patient by a genetic counselor. Three commonly used probability models were used to calculate each woman's predicted risk of harboring a germline BRCA1/2 mutation. RESULTS: Eleven of 50 patients (22% [95% confidence interval, 12-36%]) were calculated to have a > or = 10% probability of carrying a BRCA1/2 mutation by at least one mathematic model and should have been offered genetic counseling that included the discussion of genetic testing. There were considerable discrepancies between probability calculations among the three mathematic models. One of the 11 patients who was eligible for genetic testing pursued genetic counseling within 12 months of diagnosis. CONCLUSIONS: At a large academic medical center, a substantial proportion of unselected women attending a multidisciplinary clinic were found to have a > or = 10% risk of carrying a BRCA1/2 mutation. The actual number of patients eligible to receive BRCA1/2 genetic testing outweighs the number of patients seen for genetic counseling at the study institution. Finally, limited correlation was found between current predictive models. PMID- 11900217 TI - Predictive model of axillary lymph node involvement in women with small invasive breast carcinoma: axillary metastases in breast carcinoma. AB - BACKGROUND: Axillary lymph node involvement (ALNI) remains the most accurate predictive factor for recurrence risk and survival in patients with invasive breast carcinoma (IBC) and is an essential element in therapeutic decisions. However, axillary dissection (AD) is responsible for several side effects and is now discussed in small IBC. The objective of this study was to define a predictive model of ALNI by using clinical and histologic variables available before surgery. METHODS: The authors studied 795 cases of IBC (T0, T1, T2 < or = 4 cm; N0; M0) treated between 1980 and 1997 by conservative surgery and radiation therapy. All cases had axillary dissection with at least 10 lymph nodes removed. A stepwise logistic regression analysis was performed to build a predictive model of ALNI. The authors then used the jackknife resampling technique to produce unbiased estimates of the probabilities of ALNI along with their confidence intervals. RESULTS: The global ALNI rate was 25.7%. The final predictive model included clinical tumor size, location, and histologic subtype and grade as variables independently associated with ALNI. The estimated probability of ALNI varied from 6% to 45%, according to case characteristics for these variables. CONCLUSIONS: These results show that the omission of AD in surgical procedures for these tumors is debatable. Even when ALNI rates were low, the superior bounds of the confidence intervals could be high. Consequently, we do not recommend to omit AD in women whose estimated risks are higher than 25%. Women with a risk of ALNI lower than 25% could benefit from the sentinel lymph node procedure with, likewise, a limited risk of false-negative. PMID- 11900218 TI - Very early detection of RET proto-oncogene mutation is crucial for preventive thyroidectomy in multiple endocrine neoplasia type 2 children: presence of C-cell malignant disease in asymptomatic carriers. AB - BACKGROUND: Multiple endocrine neoplasia type 2 (MEN 2) is an inherited disease caused by germline mutations in the RET proto-oncogene, and is responsible for the development of endocrine neoplasia. Its prognosis is dependent on the appearance and spread of medullary thyroid carcinoma (MTC). Relatives at risk can be identified before clinical or biochemical signs of the disease become evident. METHODS: Twenty-one families with MEN 2 (16 families with MEN 2A and 5 families with MEN 2B) were studied. Peripheral blood DNA was amplified by polymerase chain reaction. DNA sequence or restriction enzyme analysis was performed to detect mutations of RET proto-oncogene exons 10, 11, and 16. Molecular analysis was carried out in all index patients as well as in 98 relatives of MEN 2A patients (60 juveniles, ages 6 months to 21 years, and 38 adults, ages 22 to 81 years) and in 13 relatives (6 juveniles ages 10 to 21 years, and 7 adults ages 41 to 66 years) from MEN 2B families. RESULTS: Molecular studies showed a mutation at codon 634, exon 11 in all MEN 2A patients. All MEN 2B patients showed an ATG to ACG (Met918Thr) mutation. In MEN 2A families, 42 out of 98 relatives were affected. Total thyroidectomy was performed in 18 juvenile carriers ages 17 months to 21 years. Histopathologic studies of the glands revealed parafollicular cell (C-cell) hyperplasia in all of these carriers, medullary thyroid carcinoma in 15 carriers, and only one carrier with lymph node metastases. CONCLUSIONS: The consistent finding of C-cell disease in all the juvenile carriers who underwent preventive thyroidectomy emphasizes the relevance of early screening in children at risk of developing MTC. The presence of MTC in the specimen of prophylactic thyroidectomy from a 17 month old girl highlights the importance of thyroidectomy as soon as the molecular diagnosis is confirmed. PMID- 11900219 TI - Prospective study of the antitumor efficacy of long-term octreotide treatment in patients with progressive metastatic gastrinoma. AB - BACKGROUND: Malignant pancreatic endocrine tumors (PETs) have a poor prognosis and existing antitumor treatments are unsatisfactory. Recent studies have shown somatostatin analogues to have antitumor growth effects in patients with malignant PETs; however, to the authors' knowledge, little information exists regarding their efficacy or effect on survival in patients with progressive malignant gastrinoma, the most common symptomatic malignant PET. The purpose of the current study was to study prospectively the efficacy, safety, and effect on survival of long-term treatment with octreotide in consecutive patients with progressive malignant gastrinoma. METHODS: Fifteen consecutive patients with malignant gastrinoma with progressive hepatic metastases were studied. All patients underwent conventional imaging studies (computed tomography scan, magnetic resonance imaging, ultrasound, and, if needed, selective angiography) and somatostatin receptor scintigraphy prior to treatment and at 3-6-month intervals while receiving treatment. The patients all were treated initially with octreotide, 200 microg every 12 hours, and at last follow-up were being maintained on long-acting release octreotide, 20-30 mg every month. Tumor size and/or number were used to classify patient responses as either no tumor response or tumor response (stabilization or decrease in size). Treatment response was correlated with tumor and clinical characteristics. RESULTS: Tumors in 8 of the 15 patients studied (53%) responded at 3 months, with 47% (7 of 15 patients) demonstrating tumor stabilization and 6% (1 of 15 patients) demonstrating a decrease in tumor size. The mean duration of response was 25.0+/-6.1 months (range, 5.5-54.1 months). Six of the eight responders were continuing to respond at the time of last follow-up. Tumor response did not correlate with any clinical parameter (e.g., tumor extent, fasting gastrin, or acid secretory rates). However, slow-growing tumors were more likely to respond prior to treatment (86% vs. 0%) (P < 0.0014). During follow-up (range, 4-8 years), 25% of the responders died compared with 71% of the nonresponders, a difference that approached statistical significance (P = 0.10). Two patients (13%) developed serious side effects that required the withdrawal of octreotide. CONCLUSIONS: Octreotide is an effective antitumor treatment in patients with progressive malignant gastrinoma. In approximately 50% of these patients octreotide has an antigrowth effect; treatment is associated with a low incidence of serious side effects compared with other antitumor treatments commonly used and, in contrast to many studies, the growth response is long-lasting. The results of the current study suggest that octreotide treatment should replace chemotherapy as the standard treatment for these patients, especially those patients with slow-growing tumors. Additional studies involving larger numbers of patients will be needed to determine a convincing effect on survival. PMID- 11900221 TI - Ten-year survival with chemotherapy and radiotherapy in patients with squamous cell carcinoma of the esophagus. AB - BACKGROUND: The effects of multimodality treatment on the survival of patients with esophageal carcinoma are unclear. The authors performed a prospective, Phase II study to assess the long-term results of chemotherapy plus radiotherapy (RT) on patients with esophageal squamous cell carcinoma. METHODS: Of 106 consecutive patients who were recruited between 1985 and 1992, 101 patients were evaluable. Cisplatin (100 mg/m2 per day) on Day 1 and fluorouracil (1000 mg/m2 per day) on Days 1-4 were given for two cycles, with concomitant RT (30 grays [Gy] in 15 fractions) over 19 days. Patients with potentially resectable tumors were then assessed for curative surgery; the other patients received two more courses of chemotherapy and further RT (20 Gy in 10 fractions). RESULTS: Of 40 patients who were candidates for surgery, 32 patients underwent surgery, and 24 patients had complete resection; 8 patients (25%) had no residual tumor in the specimen, and 12 patients (37%) had microscopic foci only. Surgical mortality was high (22%). Of 61 nonsurgical patients, 37 patients (61%) achieved complete clinical remission, and 14 patients (23%) achieved partial remission. The median survival for the entire series was 15 months (range, 1-136 months). The overall survival rate was 22% at 5 years and 12% at 10 years. At 10 years, freedom from disease progression was similar in the two groups (24%), whereas the median survival (22 months vs. 12 months) and the overall survival rates (17% vs. 9%) were better in nonsurgical patients compared with surgical patients, respectively, probably in relation to high surgical mortality. The larynx was preserved in 28% of 32 patients with cervical disease sites, with a 10-year disease free survival rate of 31%. Three deaths were attributed to nonsurgical treatments. CONCLUSIONS: Careful multidisciplinary pretreatment evaluation can identify patients who are ineligible for surgery without compromising long-term results. For patients with inoperable disease, chemoradiotherapy can produce relatively good long-term results. The combined approach without surgery can permit laryngeal preservation in a useful fraction of patients. PMID- 11900220 TI - Activation of Src kinase in primary colorectal carcinoma: an indicator of poor clinical prognosis. AB - BACKGROUND: The specific activity of the non-receptor protein tyrosine kinase, Src, is increased in the majority of colon and rectal adenocarcinomas compared to normal mucosa. However, the prognostic significance of this difference is unknown. The objective of the current study was to determine if Src activity is a marker for poor clinical prognosis in colon carcinoma patients. As Src activation leads to expression of urokinase/plasminogen activator receptor (u-PAR), expression of Src and u-PAR were correlated with patient survival. METHODS: Tumors and adjacent normal colonic mucosae from 45 patients with colorectal carcinoma were screened for Src activity by the immune complex kinase assay. Expression of u-PAR was determined by enzyme linked immunoabsorbent assay. The primary tumor-to-normal mucosa ratios of activity were compared following classification and regression tree (CART) analysis to determine the prognostic significance of elevated specific Src activity. Expression of u-PAR was correlated with Src activity. RESULTS: By CART analysis, Src activity in tumors elevated more than twofold over normal mucosa was significant. Increased Src activity significantly correlated with Dukes stage, pT and pN classification, and increased u-PAR levels (P < 0.001). Kaplan Meier analysis showed a significant association between elevated Src activity and shorter overall survival of all patients (P = 0.0004) and of Dukes Stage A-C patients (P = 0.0037). In patients who underwent curative resection, a significant correlation with a decreased disease-free survival rate was found (P < 0.0001). Multivariate analysis revealed that elevated Src activity was a prognostic parameter independent of M classification (P = 0.0125, relative risk 3.54, 95% confidence interval 1.31 - 9.76). CONCLUSIONS: Src activity is an independent indicator of poor clinical prognosis in all stages of human colon carcinoma. These data suggest that Src specific inhibitors may have a therapeutic role in inhibiting tumor progression and metastasis, and that measurement of Src activity may aid in selection of early stage patients for adjuvant therapy. PMID- 11900222 TI - Testosterone recovery following prolonged adjuvant androgen ablation for prostate carcinoma. AB - BACKGROUND: This study was conducted to describe the rate and completeness of the recovery of testosterone production following prolonged temporary androgen ablative therapy in men with prostate carcinoma undergoing curative radiation therapy. METHODS: Two-hundred and sixty-seven men treated with between 3 months and 3 years of adjuvant androgen ablation (AA) were followed at 6-month intervals following cessation of their androgen deprivation therapy. A comparative group of 518 men not undergoing AA were also followed. RESULTS: Drugs used included low dose cyproterone/stilboestrol (CPA/DES) in combination (56%) and 1 month depot (18%) and 3 month depot (25%) leutinizing hormone releasing hormone agonist (LHRHa). Seventy-nine percent of men in the current study recovered normal testosterone levels (10nmol/L), and 93% recovered levels of at least 5nmol/L. In comparison, men who had never received androgen ablative therapy showed a fall of testosterone, with 17% having sub-normal levels after 3 years. Median time to testosterone recovery was 10 months. Factors associated on multivariate analysis with delayed testosterone recovery included advanced age (P = 0.008), low pre therapy testosterone (P = 0.04), and the use of 3 month LHRHa preparations as compared with CPA/DES (P = 0.002) or 1 month LHRHa preparations (P = 0.015). The duration of drug use was not significantly associated with time to testosterone recovery. CONCLUSIONS: Long-acting LHRHa preparations appear to have a more prolonged action than previously supposed. Most men treated for up to 2 years recover normal testosterone levels after cessation of adjuvant androgen ablation, and the limited data available in the current study on patients treated for 3 years also suggests most will recover. PMID- 11900223 TI - Neuropeptides bombesin and calcitonin inhibit apoptosis-related elemental changes in prostate carcinoma cell lines. AB - BACKGROUND: Etoposide-induced apoptosis in prostate carcinoma cells is associated with changes in the elemental content of the cells. The authors previously reported that calcitonin and bombesin inhibited etoposide-induced apoptosis in these cells. In the current study, the authors investigated whether these neuropeptides block the etoposide-induced changes in elemental content. METHODS: Cells from the PC-3 and Du 145 prostate carcinoma cell lines were grown either on solid substrates or on thin plastic films on titanium electron microscopy grids, and they were exposed to etoposide for 48 hours in the absence or presence of calcitonin and bombesin. After the exposure, the cells were frozen and freeze dried, and their elemental content was analyzed by energy-dispersive X-ray microanalysis in both in the scanning electron microscope and the scanning transmission electron microscope. RESULTS: Etoposide treatment consistently induced an increase in the cellular Na concentration and a decrease in the cellular K concentration, resulting in a marked increase of the Na/K ratio and also an increase in the phosphorus:sulphur (P/S) ratio. Both bombesin and calcitonin inhibited the etoposide-induced changes in the cellular Na/K ratio, and calcitonin, but not bombesin, inhibited the changes in the P/S ratio. No significant elemental changes were found with bombesin or calcitonin alone. CONCLUSIONS: The neuropeptides bombesin and calcitonin, which inhibited etoposide induced apoptosis, also inhibited the etoposide-induced elemental changes in prostate carcinoma cells. This important fact strengthens the link between apoptosis and changes in the intracellular elemental content. This correlation provides an objective basis for the study of neuropeptide target points and may be helpful for alternative therapeutic protocols using neuropeptide inhibitors in the treatment of patients with advanced prostatic carcinoma. PMID- 11900224 TI - Loss of E-cadherin expression resulting from promoter hypermethylation in oral tongue carcinoma and its prognostic significance. AB - BACKGROUND: E-cadherin is expressed on the surface of normal epithelial cells. Loss of E-cadherin expression has been found in cancers and is postulated to facilitate tumor cell dissociation and metastasis. This study evaluated the role of promoter dense methylation in the downregulation of E-cadherin expression in oral tongue carcinoma. METHODS: E-cadherin expression of 109 oral tongue carcinomas (93 primary tumors, 7 locally recurrent tumors, and 9 metastatic lymph nodes) was evaluated by immunohistochemical staining of tumor tissues. The methylation status of the CpG islands at the promoter region of E-cadherin which flanked five HpaII (methylation sensitive restriction enzyme) digestion sites were evaluated by methylation sensitive polymerase chain reaction in 86 tumors (70 primary tumors, 7 locally recurrent tumors, and 9 metastatic lymph nodes). RESULTS: Underexpression of E-cadherin was found in 83% of primary tumors, 86% of recurrent tumors, and 89% of nodal metastases. Hypermethylated E-cadherin promoter was found in 64% of primary tumors, 71% of recurrent tumors, and 67% of nodal metastases. Downregulation of E-cadherin expression was found to be related to promoter hypermethylation. Consistently weak expression of E-cadherin by promoter hypermethylation was observed in primary tumors, their corresponding metastatic lymph nodes, and recurrent tumors. Downregulation of E-cadherin expression was a significant poor prognostic factor for survival. CONCLUSIONS: Methylation of CpG sites at the promoter region played a key role in the inhibition of E-cadherin expression in both primary oral tongue carcinomas and their corresponding recurrences and nodal metastases. The resulting downregulation of E-cadherin expression had adverse effects on the prognosis of patients who were treated by primary surgery. PMID- 11900225 TI - Expression of nucleotide excision repair genes and the risk for squamous cell carcinoma of the head and neck. AB - BACKGROUND: Phenotypic differences in the ability to repair genetic damage induced by tobacco carcinogens may reflect genetic differences in susceptibility to squamous cell carcinoma of the head and neck (SCCHN). The objective of this study was to assess the variation in baseline expression of five nucleotide excision repair genes between individuals with SCCHN and cancer free controls. METHODS: The authors conducted a hospital-based case-control study of 57 SCCHN patients and 105 cancer free controls. Using peripheral blood lymphocytes, a multiplex reverse transcriptase-polymerase chain reaction assay was used to quantitate in vitro the mRNA levels of five genes (ERCC1, XPB/ERCC3, XPG/ERCC5, CSB/ERCC6, and XPC) involved in the nucleotide excision repair pathway. RESULTS: The levels of ERCC1, XPB/ERCC3, XPG/ERCC5, and CSB/ERCC6 transcripts were lower in cases than in controls (P =0.0001, 0.096, 0.001, and 0.0001, respectively). In multivariate logistic regression analysis (adjusting for age, gender, race, smoking status, and alcohol use), low expression of ERCC1, XPB/ERCC3, XPG/ERCC5, and CSB/ERCC6 was associated with a statistically significant increased risk for SCCHN (adjusted odds ratios [95% confidence intervals] 6.42 [2.63-15.69], 2.86 [1.39-5.90], 3.69 [1.73-7.90], and 2.46 [1.19-5.09], respectively). CONCLUSIONS: Reduced expression of ERCC1, XPB/ERCC3, XPG/ERCC5, and CSB/ERCC6 is associated with a more than two-fold increased risk of SCCHN. PMID- 11900226 TI - Inducing oral immune regulation of hepatitis B virus envelope proteins suppresses the growth of hepatocellular carcinoma in mice. AB - BACKGROUND: Hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) expresses hepatitis B surface antigen (HBsAg) on its cell surface, and this may serve as a tumor-associated antigen. It was shown previously that adoptive transfer of immunity against HBsAg facilitates the suppression of experimental human HCC-expressing HBsAg in athymic mice. The authors recently showed that it was possible to augment the anti-HBV immune response through induction of oral immune regulation for HBV-associated antigens. The objective of this study was to evaluate the effect of oral immune regulation for HBV antigens on the growth of HBsAg-expressing HCC. METHODS: Recipient athymic Balb/c mice were irradiated sublethally and injected with 10(7) human hepatoma cells followed by the adoptive transfer of 2 x 10(6) splenocytes from donor mice. Four groups of donor Balb/c mice were studied: Two groups were immune modulated through oral administration of HBV antigens (HBsAg, PreS1, and Pre S2) or bovine serum albumin (BSA). Two control groups were immunized for HBsAg and fed HBV antigens or BSA. Recipient mice were followed for tumor volume and serum alpha-fetoprotein (aFP) levels. The humoral immune response was determined by measuring serum HBs antibodies. HBV specific T-cell immune modulation was assessed in vitro by HBV specific T-cell proliferation and interferon gamma (IFNgamma) ELISPOT assays as well as cytokine expression by reverse transcriptase-polymerse chain reaction assays. RESULTS: The adoptive transfer of orally immune modulated HBV splenocytes induced complete tumor suppression in recipient mice compared with control mice transplanted with nonimmune modulated cells (BSA), which showed significant tumor growth (serum aFP levels were 3.5 ng/mL and 2320.0 ng/mL, respectively). Control mice transplanted with anti-HBs immunized cells (with or without oral immune modulation) manifested similar tumor suppression (3.5 ng/mL and 0.5 ng/mL, respectively). Immunoregulation for HBV antigens augmented the HBV specific T-cell immune response, as manifested by an increase in HBV specific T-cell proliferation and IFNgamma ELISPOT assays in mice orally immune regulated with HBV proteins compared with naive mice. Tumor suppression was mediated through increased IFNgamma production in immune regulated and immunized mice. CONCLUSIONS: The induction of oral immune regulation for HBV antigens modulated the antitumor immune response, thus suppressing the growth of HCC in mice. This effect was mediated by the enhancement of anti-HBV specific T-cell immunity. PMID- 11900227 TI - HER-2/neu overexpression is rare in hepatocellular carcinoma and not predictive of anti-HER-2/neu regulation of cell growth and chemosensitivity. AB - BACKGROUND: The overexpression of HER-2/neu oncogene has been implicated in the development and modulation of many types of cancer. However, whether HER-2/neu overexpression plays a similar role in hepatocellular carcinoma (HCC) has not been determined. METHODS: Tissue specimens from 36 HCC patients who had been enrolled in 3 separate prospective clinical trials of systemic chemotherapy were studied by immunohistochemical staining. A polyclonal antibody (A0485; DAKO, Copenhagen, Denmark) against HER-2/neu and a horseradish peroxidase-based visualization system (Envision+, DAKO) was used. Scoring criteria was in accordance with the manufacturer's guidelines. Twelve HCC cell lines were examined for HER-2/neu overexpression by Western blotting. Single-agent growth regulatory activity of the anti-HER-2/neu antibody, trastuzumab (Herceptin; Genentech, South San Francisco, CA), and its combinative cytotoxicity with chemotherapeutic agents (doxorubicin, gemcitabine, cisplatin, irinotecan) were determined by a tetrazolium-based colorimetric assay (MTT test). RESULTS: All but one of the HCC tumor tissues were negative for HER-2/neu expression. The only patient with positive HER-2/neu expression was a 57-year-old male patient who achieved stabilization of disease for 2 months after chemotherapy. Eight of the 35 patients with negative HER-2/neu expression had had partial remission after chemotherapy (P = 0.78). Only one (Tong cells) of the 12 HCC cell lines had a significant level of HER-2/neu expression. However, trastuzumab up to 10 microg/mL had no discernible growth inhibitory or chemosensitizing effect on Tong cells or any other cell lines. CONCLUSIONS: HER-2/neu overexpression is rare in human HCC tissues, and anti-HER-2/neu regulation appears to play little role in the treatment of this tumor. PMID- 11900228 TI - Matrix metalloproteinase-7 expression and biologic aggressiveness of cholangiocellular carcinoma. AB - BACKGROUND: Matrix metalloproteinase-7 (MMP-7) recently has been reported to play a role in tumor cell invasion. The objective of the current study was to examine the expression of MMP-7 in patients with cholangiocellular carcinoma. METHODS: Twenty-six patients underwent resection of cholangiocellular carcinoma, leaving no macroscopic evidence of residual tumor. Immunostaining was performed to evaluate the relation between MMP-7 expression and clinicopathologic features and patient prognosis. The immunostaining pattern of the tumor cells for MMP-7 was classified as negative (-) (n = 9), positive (+) (n = 11), or strongly positive (++) (n = 6). Western blot analysis was performed to investigate the expression of active or latent forms in cancerous and noncancerous lesions in four patients. RESULTS: The survival rates in patients with MMP-7 expression judged to be (-), (+), and (++) were 75%, 80%, and 0%, respectively, at 1 year, and 47%, 24%, and 0%, respectively, at 3 years. The survival rate of MMP-7 (++) patients was significantly lower than that of MMP-7 (+) patients (P = 0.003) and MMP (-) patients (P = 0.008). At last follow-up, 3 patients in the MMP-7 (-) group had survived for > 5 years. Western blot analysis demonstrated that there were two types of cholangiocellular carcinoma: those producing both latent and active MMP 7 and those producing only the latent form. CONCLUSIONS: Although the results of the current study are based on a small number of patients, they suggest that MMP 7 expression is a significant prognostic factor in patients with cholangiocellular carcinoma and that cholangiocellular carcinoma demonstrating strongly positive expression of MMP-7 on immunostaining may have a higher malignant potential compared with that showing negative or positive expression of MMP-7. PMID- 11900229 TI - Combined intraarterial 5-fluorouracil and subcutaneous interferon-alpha therapy for advanced hepatocellular carcinoma with tumor thrombi in the major portal branches. AB - BACKGROUND: The prognosis of hepatocellular carcinoma (HCC) invading into the major branches of the portal vein (Vp3) is extremely poor. METHODS: Eleven consecutive patients with HCC and Vp3 were treated with 2-6 cycles of a "basic" combination therapy consisting of continuous arterial infusion of 5-fluorouracil (450-500 mg/day, for the initial 2 weeks) and subcutaneous injection of interferon-alpha (5 million international units, 3 times/week, 4 weeks). In the first 3 patients, methotrexate (90 mg/day 1 of every week), cisplatin (10 mg/day), and leucovorin (30 mg/days 2 and 3 of every week) also were administered for the initial 2 weeks ("full" regimen). RESULTS: In 8 (73%) of 11 patients, an objective response (complete response [CR] or partial response [PR]) was observed with marked regression of tumor and decrease in tumor markers. The use of the full regimen was associated with objective response in all patients; instead, they developed thrombocytopenia or leukopenia. In the subsequent 8 patients with basic regimen, 5 patients showed CR (2 cases) or PR (3 cases; objective response rate, 63%), and leukopenia was observed only in 1 patient. CONCLUSIONS: Simple combination therapy with subcutaneous interferon-alpha and intraarterial 5 fluorouracil therefore is a promising treatment modality for intractable HCC with Vp3. PMID- 11900230 TI - Percutaneous tumor ablation with radiofrequency. AB - BACKGROUND: Radiofrequency thermal ablation (RFA) is a new minimally invasive treatment for localized cancer. Minimally invasive surgical options require less resources, time, recovery, and cost, and often offer reduced morbidity and mortality, compared with more invasive methods. To be useful, image-guided, minimally invasive, local treatments will have to meet those expectations without sacrificing efficacy. METHODS: Image-guided, local cancer treatment relies on the assumption that local disease control may improve survival. Recent developments in ablative techniques are being applied to patients with inoperable, small, or solitary liver tumors, recurrent metachronous hereditary renal cell carcinoma, and neoplasms in the bone, lung, breast, and adrenal gland. RESULTS: Recent refinements in ablation technology enable large tumor volumes to be treated with image-guided needle placement, either percutaneously, laparoscopically, or with open surgery. Local disease control potentially could result in improved survival, or enhanced operability. CONCLUSIONS: Consensus indications in oncology are ill-defined, despite widespread proliferation of the technology. A brief review is presented of the current status of image-guided tumor ablation therapy. More rigorous scientific review, long-term follow-up, and randomized prospective trials are needed to help define the role of RFA in oncology. PMID- 11900231 TI - A prospective study of pulmonary function in patients treated with paclitaxel and carboplatin. AB - BACKGROUND: Adverse effects of paclitaxel and carboplatin have been well described; however, pulmonary toxicity after patients receive this regimen has not been investigated extensively. METHODS: To clarify this issue, 33 consecutive patients who were treated with paclitaxel and carboplatin underwent prospective evaluation of respiratory function, which included pulmonary symptoms, pulmonary function tests (PFTs), arterial blood gas levels, and radiographic studies. Assessment was performed before and after completion of chemotherapy in all patients. Patients with substantial declines in PFTs, defined as a decline > or = 20 percent in forced expiratory volume in 1 second (FEV1), total lung capacity (TLC), or diffusion capacity for carbon monoxide (DLCO), were reassessed 5 months later. RESULTS: After chemotherapy, there were no significant changes in forced vital capacity (FVC; 111%+/-21% of the predicted value before chemotherapy vs. 111+/-20% of the predicted value after chemotherapy), FEV1 (108%+/-24% of the predicted value before chemotherapy vs. 107%+/-22% of the predicted value after chemotherapy), FEV1/FVC ratio (79%+/-8% before chemotherapy vs. 78%+/-6% after chemotherapy), alveolar volume (VA; 95%+/-14% of the predicted value before chemotherapy vs. 96%+/-14% of the predicted value after chemotherapy), or TLC (96%+/-14% of the predicted value before chemotherapy vs. 97%+/-13% of the predicted value after chemotherapy). In contrast, there was a significant decline in DLCO (101%+/-20% of the predicted value before chemotherapy vs. 96+/-21% of the predicted value after chemotherapy; P < 0.05). Arterial blood gas levels did not change after treatment. No patient had decreased FEV1 or TLC levels by > or = 20%, whereas 4 of 33 patients (12%) exhibited a substantial decline (> or = 20%) in DLCO that persisted 5 months after treatment (DLCO at baseline, immediately after chemotherapy, and 5 months after the completion of chemotherapy, respectively: 99%+/-36% of the predicted value vs. 75%+/-28% of the predicted value vs. 74%+/-31% of the predicted value; P < 0.05). None of the 33 patients developed respiratory symptoms or had radiologic signs suggestive of lung toxicity. Among the various risk factors examined, baseline DLCO and FEV1 levels were associated with changes in DLCO post-treatment. CONCLUSIONS: This prospective analysis showed that the combination of paclitaxel with carboplatin induced an isolated decrease in DLCO level in the absence of clinical or radiologic evidence of toxicity. Further studies are needed to clarify whether this reduction in DLCO is predictive of subsequent pulmonary impairment. PMID- 11900232 TI - A phase II study of neoadjuvant biochemotherapy for stage III melanoma. AB - BACKGROUND: Phase II studies of biochemotherapy (combining interleukin-2, interferon-alpha, and multiagent chemotherapy) have reported high response rates and a significant number of durable complete responses in patients with metastatic melanoma. METHODS: A pilot Phase II study was performed to explore the safety and activity of neoadjuvant biochemotherapy in patients with Stage III melanoma. Forty-eight patients were enrolled between April 1996 and May 1999. The median age of the patients was 46 years (range, 19-70 years). Two cycles of biochemotherapy were administered prior to and after complete lymph node dissection. Each cycle was comprised of cisplatin, 20 mg/m2 intravenously (i.v.), on Days 1-4; vinblastine, 1.6 mg/m2 i.v., on Days 1-4; dacarbazine, 800 mg/m2 i.v., on Day 1; interleukin-2, 9 x 10(6) IU/m2/day i.v. over 24 hours, on Days 1 4; and interferon-alpha, 5 x 10(6) IU/m2/day subcutaneously, on Days 1-5, every 3 weeks. Twelve patients did not have measurable disease. All patients were evaluable for toxicity and survival. RESULTS: Clinical responses were observed in 14 of 36 patients (38.9%) with measurable disease, including 13 partial responses (36.1%) and 1 complete response (2.8%). Complete pathologic responses were noted in 4 patients (11.1%). Toxicity, although severe, was manageable and typically short-lived. There were no treatment-related deaths reported. At a median follow up of 31 months, 38 of the 48 patients (79.2%) were alive and 31 patients (64.6%) remained free of disease progression. CONCLUSIONS: Neoadjuvant biochemotherapy appears to have promising activity in patients with Stage III melanoma. A larger multicenter study currently is underway to explore this approach further. PMID- 11900233 TI - Immunotherapy for nonmelanoma skin cancer: does it have a future? AB - BACKGROUND: Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) of the skin are the most common malignancies in the white human population, accounting for greater than 95% of nonmelanoma skin cancers (NMSCs). Current data show an increasing incidence of NMSC in recent decades. Although the mortality is low, this cancer group is associated with substantial morbidity. Multiple treatment modalities are available for NMSC, with surgery being a "cornerstone" of current therapy approaches. However, in patients with multiple lesions or in cases of tumors on critical locations, disfigurement and the disease recurrence may represent a serious problem associated with the surgical treatment. The purpose of this study was to review and analyze whether NMSC could represent a target for immune therapy, evaluating the aspects of the availability of tumor antigens and the existence of tumor specific immune response, including a summary of the major clinical studies dealing with immunotherapy for NMSC. METHODS: The authors have reviewed the available medical literature on NMSC, with a focus on tumor immunology and associated abnormalities, as well as immunotherapy-based treatment trials. RESULTS: The major advantage of NMSCs is that they arise from the skin, which makes them easily detectable and treatable. Furthermore, these tumors posses all the prerequisites, i.e., the presence of tumor-associated antigens as well as the tumor specific immune response, needed for immune intervention. This also was confirmed in various studies demonstrating clinical efficacy of cytokines and other immune response modifiers. CONCLUSIONS: In addition to clinical cure, by activating and stimulating patient's immune resources this therapeutic option may be a "silver bullet," providing a long-term protective immunity against initial tumor. PMID- 11900234 TI - Hyperthermic intraoperative intraperitoneal chemotherapy with cisplatin and doxorubicin in patients who undergo cytoreductive surgery for peritoneal carcinomatosis and sarcomatosis: phase I study. AB - BACKGROUND: Hyperthermic intraperitoneal intraoperative chemotherapy (HIIC) combined with cytoreductive surgery (CS) has been proposed as a new multimodal treatment mainly for carcinomatosis of gastrointestinal origin. To evaluate whether this regimen could be used for other tumor types, the authors conducted a Phase I study on HIIC with doxorubicin and cisplatin in patients with peritoneal carcinomatosis or sarcomatosis. PATIENTS AND METHODS: Thirty-one patients with peritoneal carcinomatosis or sarcomatosis (PCS) were enrolled for the study. After completion of CS, HIIC was administered with drug doses that were increased for each consecutive cohort following a three-patient cohort scheme. Thereafter, the accrual was stopped when Grade 4 locoregional or systemic toxicity was observed. The maximum tolerated dose (MTD) was considered the dose in the previous triplet. Drug pharmacokinetics and procedure costs also were analyzed. RESULTS: After CS, residual tumors were not present or measured less than or equal to 3 mm (in dimension) in all cases. Maximum tolerated dose was 15.25 and 43.00 mg L(-1) for doxorubicin and cisplatin, respectively. The perfusate/plasma area under the curve ratios were favorable for both drugs, at 162+/-113 and 20.6+/-6.0, respectively, for doxorubicin and cisplatin. Doxorubicin levels in the peritoneum were higher than in tumor or normal tissue samples. There were no postoperative deaths. Surgery-related complications were observed in 25% of cases. Findings at cost analysis showed that the length of stay in the operation room and intensive care unit were the major cost drivers. CONCLUSIONS: Cytoreductive surgery combined with HIIC is an expensive but feasible therapeutic approach for locally advanced abdominal tumors. Because our preliminary findings for local disease control are encouraging, a Phase II study is now advisable to verify the activity of this promising treatment. PMID- 11900236 TI - Self-assessment in cancer patients referred to palliative care: a study of feasibility and symptom epidemiology. AB - BACKGROUND: Research in palliative care is considered difficult due to the poor health of patients. However, patient-provided data are essential for a thorough description of patient symptomatology and for the evaluation of care. METHODS: The authors examined the feasibility of a questionnaire-based study using the European Organization for Research and Treatment of Cancer quality-of-life instrument EORTC QLQ-C30, the Edmonton Symptom Assessment System (ESAS), and the Hospital Anxiety and Depression Scale (HADS) in cancer patients who were receiving palliative care. This report describes the symptomatology of participating patients and examines differences in symptomatology between patients in three palliative care functions: inpatient, outpatient, and palliative home care. RESULTS: Of 267 eligible patients who were referred to a department of palliative medicine, initial self-assessment questionnaires were obtained from 176 patients (65.9%). The 91 nonparticipants were older and had lower Karnofsky Performance status (KPS) values than the participants. Almost all participating patients suffered from impaired role function and physical function and had high levels of pain, fatigue, and other symptoms. According to the HADS, 47% of patients suffered from depression. Outpatients had better scores than inpatients and patients in palliative home care for physical function, role function, cognitive function, depression, and inactivity. CONCLUSIONS: It is possible to carry out a questionnaire-based study of symptomatology in consecutive cancer patients in palliative care, achieving rather complete data from the participants. The symptomatology in these patients was very pronounced. The questionnaires were able to detect clinically important differences between places of service. PMID- 11900235 TI - The American Cancer Society Cancer Prevention Study II Nutrition Cohort: rationale, study design, and baseline characteristics. AB - BACKGROUND: Large-scale, prospective cohort studies have played a critical role in discovering factors that contribute to variability in cancer risk in human populations. Epidemiologists and volunteers at the American Cancer Society (ACS) were among the first to establish such cohorts, beginning in the early 1950s and continuing through the present, and these ACS cohorts have made landmark contributions in many areas of epidemiologic research. METHODS AND RESULTS: The Cancer Prevention Study II Nutrition Cohort was established in 1992 and was designed to investigate the relation between diet and other lifestyle factors and exposures and the risk of cancer, mortality, and survival. The cohort includes over 84,000 men and 97,000 women who completed a mailed questionnaire in 1992. New questionnaires are sent to surviving cohort members every other year to update exposure information and to ascertain new occurrences of cancer; a 90% response rate was achieved for follow-up questionnaires in 1997 and 1999. Reported cancers are verified through medical records, registry linkage, or death certificates. The cohort is followed actively for all cases of incident cancer and for all causes of death. Through a collaborative effort among ACS national and division staff, volunteers, and the American College of Surgeons, blood samples were collected from a subgroup of 40,000 cohort members and are in storage at a central repository for future investigation of dietary, hormonal, genetic, and other factors and cancer risk. Collection of DNA samples from buccal cells in an additional 50,000 cohort members is underway currently and will be completed in 2002. CONCLUSIONS: This new cohort of both men and women promises to be particularly valuable for the study of cancer occurrence, mortality, and survival as they relate to obesity and weight change, physical activity at various points in life, vitamin supplement use, exogenous hormone use, other medications (such as aspirin and nonsteroidal anti-inflammatory drugs) and cancer screening modalities. PMID- 11900237 TI - Lifestyle interference and emotional distress in family caregivers of advanced cancer patients. AB - BACKGROUND: Providing end-of-life care at home to a family member with advanced cancer can have a negative impact on the emotional well-being of the family caregiver. The current study examined the impact of providing care on lifestyle and emotional well-being in a sample of caregivers to patients with advanced cancer. The mediation of lifestyle interference between the amount of care provided and emotional distress was specifically examined. METHODS: Forty-four family caregivers participated in a structured quantitative interview. Lifestyle interference was assessed by the Caregiving Impact Scale, amount of care provided was assessed by the Caregiver Assistance Scale, and emotional distress was assessed by the Profile of Mood States-Short Form. Pearson and partial correlations tested whether lifestyle interference mediated the relationship between caregiving assistance and emotional distress. Regression analyses determined overall correlates of emotional distress. RESULTS: Three criteria, required to substantiate mediation, were met for total mood disturbance and the depression and tension subscales. An overall regression model identified education level and lifestyle interference to be significant and unique correlates of emotional distress. CONCLUSIONS: The current results suggest that caregivers experience increased emotional distress, regardless of the amount of care provided, when limited in their ability to participate in valued activities and interests. In addition, caregivers with less than a high school education experience more emotional distress. Therefore, helping caregivers maintain valued aspects of their lifestyle should be an important element of home care. PMID- 11900238 TI - Fatigue in cancer patients compared with fatigue in the general United States population. AB - BACKGROUND: Although fatigue is a common symptom among cancer patients, it is also a common experience in the general, healthy population. Its universality has made it difficult to appreciate whether the fatigue experienced by patients with cancer is distinguishable from the fatigue experienced by the general population. Because the etiology of fatigue is multifactorial, it also has been difficult to appreciate fully the relative contribution of anemia to cancer-related fatigue. METHODS: To address this issue, responses to a brief, standardized set of 13 questions from the Functional Assessment of Chronic Illness Therapy (FACIT) Measurement System were compared across three groups: anemic cancer patients (n = 2369 patients), nonanemic cancer patients (n = 113 patients), and the general United States population (n = 1010 persons). RESULTS: Fatigue scores of the anemic cancer patients (at both baseline and upon completion of anemia therapy) were significantly worse compared with the scores of nonanemic cancer patients that, in turn, were worse compared with the scores of the general United States population (P < 0.001). Score distributions were quite distinct for these three groups. Within the group of anemic cancer patients, the degree of anemia (mild, moderate, or severe) also was predictive of the degree of fatigue (P < 0.001), although the distributions were not dramatically distinct. CONCLUSIONS: Although anemia is clearly a factor that contributes to the severity of disease-related fatigue among cancer patients, hemoglobin levels explain only part of the difference compared with fatigue among the general United States population. The distinct distributions of fatigue scores of anemic cancer patients compared with the general United States population and the substantial sample sizes of these two groups enabled a discriminant analysis approach that allowed the differentiation of anemic cancer patients from the general population with high sensitivity (0.92) and reasonable specificity (0.69). Thus, although fatigue is a symptom most anyone can relate to, the fatigue of cancer patients, particularly those who are anemic, is decidedly worse. Interventions targeting this common and life-disrupting symptom likely would be of considerable value to patients with cancer. PMID- 11900239 TI - Physical exercise and immune system function in cancer survivors: a comprehensive review and future directions. AB - BACKGROUND: There are a limited number of interventions for cancer survivors following completion of primary therapy that might reduce the risk of cancer recurrence and/or secondary malignancies and increase survival times. It has been proposed that physical exercise may be beneficial by enhancing the anticancer immune system response. The purpose of the current article is to: 1) briefly describe the immune system response to tumors, 2) discuss the impact of anticancer therapy on immune system function in cancer survivors, 3) provide a systematic and comprehensive review of the extant literature examining physical exercise and immune system function in cancer survivors, and 4) offer a critical analysis of this literature and outline directions for future research. METHODS: A comprehensive literature search up to March 2001 identified empirical articles that examined the effects of physical exercise training on immune system function in cancer survivors from CD-ROM database searches and manual searches. RESULTS: To the authors' knowledge, six empirical studies published between 1994 and 2000 have examined physical exercise and immune system function in cancer survivors. Overall, four out of six studies reported statistically significant improvements in a number of cancer-related immune system components as a result of exercise. However, there are several limitations that must be considered when interpreting the findings of these studies. These limitations involve the samples, designs, physical exercise interventions, physical fitness assessments, and immunologic assessments. CONCLUSIONS: Additional research is needed to determine if physical exercise in cancer survivors may reduce the risk of cancer recurrence and secondary malignancies and increase survival times. PMID- 11900240 TI - Histopathologic grading of medulloblastomas: a Pediatric Oncology Group study. AB - BACKGROUND: Medulloblastomas are small cell embryonal tumors of the cerebellum found predominantly in children, only slightly more than half of whom survive. Predicting favorable outcome has been difficult, and improved stratification clearly is required to avoid both undertreatment and overtreatment. Patients currently are staged clinically, but no pathologic staging system is in use. Two rare subtypes at extreme ends of the histologic spectrum, i.e., medulloblastomas with extensive nodularity and large cell/anaplastic medulloblastomas, are associated with better and worse clinical outcomes, respectively. However, there is little data about correlations between histologic features and clinical outcome for most patients with medulloblastomas that fall between these histologic extremes of nodularity and anaplasia. Therefore, the authors evaluated the clinical effects of increasing anaplasia and nodularity in a large group of children with medulloblastomas, hypothesizing that increasing nodularity would predict better clinical outcomes and that increasing anaplasia would presage less favorable results. METHODS: Medulloblastomas from 330 Pediatric Oncology Group patients were evaluated histologically with respect to extent of nodularity, presence of desmoplasia, grade of anaplasia, and extent of anaplasia. Pathologic and clinical data were then compared using Kaplan-Meier and log-rank analyses. RESULTS: Increasing grade of anaplasia and extent of anaplasia were associated strongly with progressively worse clinical outcomes (P < 0.0001 for both). Significant anaplasia (moderate or severe) was identified in 24% of medulloblastoma specimens. Neither increasing degrees of nodularity nor desmoplasia were associated significantly with longer survival. CONCLUSIONS: Moderate anaplasia and severe anaplasia were associated with aggressive clinical behavior in patients with medulloblastomas and were detected in a significant number of specimens (24%). Pathologic grading of medulloblastomas with respect to anaplasia may be of clinical utility. PMID- 11900241 TI - Survival after recurrence of Ewing tumors: the St Jude Children's Research Hospital experience, 1979-1999. AB - BACKGROUND: Despite improved therapies, 30-40% of patients with Ewing tumors (ET) experience recurrence and have a poor prognosis. The authors analyzed factors prognostic of survival in patients with recurrent ET. METHODS: The authors assessed the relation between postrecurrence survival (PRS) and demographic, disease, and treatment factors in 71 patients who experienced recurrent ET after treatment on one of three consecutive institutional protocols. RESULTS: Thirty four patients (47.9%) had distant recurrence, 25 patients (35.2%) had local recurrence, and 12 patients (16.9%) had both distant and local recurrence at a median of 1.7 years after diagnosis. The probability of 5-year PRS (+/- 1 standard error) was 17.7%+/-4.5%. Recurrence > or = 2 years after diagnosis predicted a significantly better outcome (5-year PRS, 34.9%+/-8.5%) compared with earlier recurrence (5.0%+/-2.8%; P < 0.001). Patients who had both local and distant recurrence fared worse (5-year PRS, 12.5%+/-8.3%) compared with patients who had local recurrence alone (21.7%+/-7.8%) or distant recurrence alone (17.6+/ 6.1%). Among patients with local recurrence alone, those who underwent salvage with radical surgery had significantly higher 5-year PRS estimates (31.4%+/ 11.6%) compared with the other patients (9.1%+/-6.1%; P = 0.023). Pulmonary irradiation significantly improved the outcomes of patients with isolated pulmonary recurrence (5-year PRS estimate, 30.3%+/-12.5% vs. 16.7%+/-10.8%, respectively; P = 0.018). CONCLUSIONS: Although outcomes are generally poor after patients experience recurrence of ET, certain patient groups differ appreciably in their likelihood of survival. Patients who experience recurrence > or = 2 years after diagnosis and patients who have local recurrence that can be treated with radical surgery and intensive chemotherapy have the most favorable outcomes. PMID- 11900242 TI - Pathologic features of superficial esophageal squamous cell carcinoma with lymph node and distal metastasis. AB - BACKGROUND: Endoscopic mucosal resection (EMR) is a less invasive localized treatment for patients with esophageal carcinoma. However, indications for EMR use in cases of superficial esophageal carcinoma are controversial. The authors evaluated histopathologic risk factors for lymph node metastasis and recurrence. METHODS: In the specimens resected, the authors examined depth, the superficial area and the area attached to or infiltrating the lamina muscularis mucosa. RESULTS: The authors found that the superficial area and the attached or infiltrated area reflected the depth of the tumor. However, there was a recurrence of esophageal carcinoma even in m3 cases attached only to the lamina muscularis mucosa. CONCLUSIONS: The authors concluded that ml and m2 esophageal carcinoma had almost no risk of lymph node metastasis and recurrence no matter how extensive the superficial area. In addition, sm2 and sm3 carcinoma have a high frequency of lymph node metastasis and recurrence. M3 and sm1 carcinoma run the risk of lymph node metastasis and recurrence however small the superficial area and the area attached to or infiltrating the lamina muscularis mucosa. Treatment strategies for patients with superficial esophageal carcinoma, including EMR, should take the above findings into account. PMID- 11900243 TI - Prognostic factors for patients with small cell lung carcinoma. PMID- 11900244 TI - Beyond randomized controlled trials: organized mammographic screening substantially reduces breast carcinoma mortality. PMID- 11900245 TI - Beyond randomized controlled trials: organized mammographic screening substantially reduces breast carcinoma mortality. PMID- 11900246 TI - Beyond randomized controlled trials: organized mammographic screening substantially reduces breast carcinoma mortality. PMID- 11900247 TI - Beyond randomized controlled trials: organized mammographic screening substantially reduces breast carcinoma mortality. PMID- 11900248 TI - Colorectal cancer prevention and treatment by inhibition of cyclooxygenase-2. AB - Population-based studies have established that long-term intake of non-steroidal anti-inflammatory drugs (NSAIDs), compounds that inhibit the enzymatic activity of cyclooxygenase (COX), reduces the relative risk for developing colorectal cancer. These studies led to the identification of a molecular target, COX-2, that is involved in tumour promotion during colorectal cancer progression. Recent studies in humans indicate that therapy with specific COX-2 inhibitors might be an effective approach to colorectal cancer prevention and treatment. PMID- 11900249 TI - How nucleotide excision repair protects against cancer. AB - Eukaryotic cells can repair many types of DNA damage. Among the known DNA repair processes in humans, one type--nucleotide excision repair (NER)--specifically protects against mutations caused indirectly by environmental carcinogens. Humans with a hereditary defect in NER suffer from xeroderma pigmentosum and have a marked predisposition to skin cancer caused by sunlight exposure. How does NER protect against skin cancer and possibly other types of environmentally induced cancer in humans? PMID- 11900250 TI - The development of androgen-independent prostate cancer. AB - The normal prostate and early-stage prostate cancers depend on androgens for growth and survival, and androgen ablation therapy causes them to regress. Cancers that are not cured by surgery eventually become androgen independent, rendering anti-androgen therapy ineffective. But how does androgen independence arise? We predict that understanding the pathways that lead to the development of androgen-independent prostate cancer will pave the way to effective therapies for these, at present, untreatable cancers. PMID- 11900252 TI - APC, signal transduction and genetic instability in colorectal cancer. AB - Colorectal cancer arises through a gradual series of histological changes, each of which is accompanied by a specific genetic alteration. In general, an intestinal cell needs to comply with two essential requirements to develop into a cancer: it must acquire selective advantage to allow for the initial clonal expansion, and genetic instability to allow for multiple hits in other genes that are responsible for tumour progression and malignant transformation. Inactivation of APC--the gene responsible for most cases of colorectal cancer--might fulfil both requirements. PMID- 11900253 TI - Rescuing the function of mutant p53. AB - One protein--p53--plays nemesis to most cancers by condemning damaged cells to death or quarantining them for repair. But the activity of p53 relies on its intact native conformation, which can be lost following mutation of a single nucleotide. With thousands of such mutations identified in patients, how can a future cancer drug buttress this fragile protein structure and restore the cell's natural defence? PMID- 11900254 TI - Cancer genetics: from Boveri and Mendel to microarrays. AB - The human genome has now been sequenced, a century after the re-discovery of Mendel's Laws, and the publication of Theodor Boveri's chromosomal theory of heredity. Tracing the historical landmarks of cancer genetics from these early days to the present time not only gives us an appreciation of how far we have come, but also emphasizes the challenges that we face if we are to unravel the genetic basis of hereditary and sporadic cancers in the next century. PMID- 11900255 TI - Tobacco and the global lung cancer epidemic. AB - Tobacco is the world's single most avoidable cause of death. The World Health Organization has calculated that the 5.6 trillion cigarettes smoked per year at the close of the twentieth century will cause nearly 10 million fatalities per year by 2030. Lung cancer is the most common tobacco-related cause of cancer mortality, with one case being produced for every 3 million cigarettes smoked. How was the global lung cancer epidemic recognized, and what can we expect in the future? PMID- 11900256 TI - Current status of rehabilitation medicine in Asia: a report from New Millennium Asian Symposium on Rehabilitation Medicine. AB - With the aim of promoting rehabilitation medicine in Asian countries, where the number of persons with disability occupies a significant proportion in the world, New Millennium Asian Symposium on Rehabilitation Medicine was held in February 2001 in Tokyo, under the sponsorship of the Japanese Association of Rehabilitation Medicine. Twenty-three guest speakers from 14 Asian countries and regions participated in the 2-day meeting. With a structured questionnaire that was sent to the participants beforehand, demographic data related to rehabilitation practice and information on training and certification in rehabilitation medicine in the participating countries were collected, and presented at the meeting. Based on these data, the current status of rehabilitation medicine in Asia was summarized. The symposium marked an important step forward for the promotion of rehabilitation medicine in Asia. PMID- 11900257 TI - Reorganization of equilibrium and movement control strategies after total knee arthroplasty. AB - This work was aimed at identifying changes in posturomotor control strategies in patients with unilateral total knee arthroplasty. Using kinetic and kinematic data, a previous study had revealed that, during a side step, patients with unilateral knee arthritis showed a shortened monopodal phase and a lengthened postural phase when the affected leg was the supporting one. It was expected that these strategies would be modified after undergoing total knee arthroplasty. Postoperatively the durations of the monopodal phase and of the postural phase became similar when the operated limb was supporting and when the sound limb was supporting. Concerning the upper body movements, the same asymmetrical results as before surgery were observed. Hence, patients with total knee arthroplasty exhibit posturomotor strategies which, although they become close to normal, remain asymmetrical. The durations of the monopodal and of the postural phases could be considered to assess the results of total knee arthroplasty. PMID- 11900251 TI - Putting tumours in context. AB - The interactions between cancer cells and their micro- and macroenvironment create a context that promotes tumour growth and protects it from immune attack. The functional association of cancer cells with their surrounding tissues forms a new 'organ' that changes as malignancy progresses. Investigation of this process might provide new insights into the mechanisms of tumorigenesis and could also lead to new therapeutic targets. PMID- 11900258 TI - Inter-rater reliability and validity of the stroke rehabilitation assessment of movement (stream) instrument. AB - The Stroke Rehabilitation Assessment of Movement (STREAM) instrument is used to measure motor and mobility problems in patients who have experienced a stroke. The purposes of the study were to examine the interrater reliability, concurrent and convergent validity of the STREAM instrument in stroke patients. Fifty-four stroke patients participated in the study. For the purpose of interrater reliability, the STREAM instrument was administered by two raters on each patient within a 2-day period. Validity was assessed by comparing the patients' scores on the STREAM instrument with those obtained from the other well-established measures. Weighted kappa statistics for inter-rater agreement on scores for individual items ranged from 0.55 to 0.94. The intraclass correlation coefficient for the total score was 0.96 indicating very high inter-rater reliability. The intraclass correlation coefficients were also very high in each of the subscales. The total STREAM score was moderately to highly associated with the score of the Barthel Index and Fugl-Meyer motor assessment scale, rho = 0.67, and 0.95, respectively. The STREAM subscale scores were closely associated with scores of the other well-validated measures. Our results demonstrate that consistent and valid information can be obtained from the STREAM instrument and support its use in the value of the STREAM evaluation of motor and mobility recovery in persons who have experienced a stroke. PMID- 11900259 TI - Effects of functional electrical stimulation training for six months on body composition and spasticity in motor complete tetraplegic spinal cord-injured individuals. AB - The effect of functional electrical stimulation (FES) training on body composition, assessed by computed tomography, and the effect of spasticity, assessed by both objective and subjective measures, are evaluated. Fifteen motor complete spinal-cord-injured men participated in the study. Eight of the 15 subjects undertook FES cycling 3 times weekly for 6 months. Whole body computed tomography scans evaluated changes in body composition. Simultaneous Modified Ashworth Scale and electromyography (EMG) measurements, resistive torque (Kin Com) and EMG measurements, and self-ratings with Visual Analogue Scale during four consecutive days were used to evaluate changes in spasticity. Lower extremity muscle volume increased by an average of 1300 cm3 (p < 0.001) in the training group compared to the control group, who experienced no change. Otherwise no changes in body composition were seen. Significant correlations (Spearman) were found between individual EMG activity recordings and movement provoked Modified Ashworth Scale ratings in 26% of the test situations, irrespective of group and time. The objective and subjective evaluation of movement-provoked passive (viscoelastic) and active (spasticity-related) resistance remained unchanged. PMID- 11900260 TI - Behaviour-focused pain coping: consistency and convergence to work capability of the swedish version of the chronic pain coping inventory. AB - The aim was to study the psychometric properties of the Swedish version of the Chronic Pain Coping Inventory. The material consisted of a group of 100 subjects recruited from a large population study. Pain status and the absence of pain related sick leave during the previous year conditioned inclusion. Another group comprised 160 patients on the long-term sick list and who had been referred to a multidisciplinary pain clinic for evaluation. The psychometric properties in terms of internal consistency of the scales were good or very good for all scales of behaviour-focused pain coping. Use of the strategies "Guarding", "Resting", "Asking for assistance", "Relaxation", "Task persistence", "Coping self statements" and "Seeking social support" was significantly related to vocational capability. "Guarding". "Asking for assistance", "Relaxation", "Exercise and stretch" and "Coping self-statements" increased in parallel to increasing pain from localized to intermediate or widespread. No gender difference was found in cases reporting more pronounced pain. PMID- 11900261 TI - Exercise training in patients with end-stage renal disease on hemodialysis: comparison of three rehabilitation programs. AB - Functional capacity of end-stage renal disease patients is dramatically impaired. Although exercise training programs appear to have beneficial morphological, functional and psychosocial effects in end-stage renal disease patients on hemodialysis (HD), the adherence rate is high. The purpose of this study was to compare the effects of three modes of exercise training on aerobic capacity and to identify the most favourable, efficient and preferable to patients on HD with regard to functional improvements and participation rate in the programs. Fifty eight volunteer patients were screened for low-risk status and selected from the dialysis population. The 48 patients who completed the study protocol were randomly assigned either to one of the three training groups or to a control group. Sixteen of them (Group A - mean age 46.4+/-13.9 years) completed a 6-month supervised outpatient exercise renal rehabilitation program consisting of three weekly sessions of aerobic and strengthening training on the non-dialysis days; 10 (Group B - mean age 48.3+/-12.1 years) completed a 6-month exercise program during HD; 10 (Group C - mean age 51.4+/-12.5 years) followed an unsupervised moderate exercise program at home, and 12 patients (Group D-mean age 50.2+/-7.9 years) were used as patient controls. The level of anemia, the medications and the HD prescription remained stable during the study. Fifteen sex- and age matched sedentary individuals (Group E - mean age 46.9+/-6.4 years) comprised a healthy control group for baseline data. All subjects at the beginning and end of the study underwent clinical examination, laboratory tests and a treadmill exercise test to fatigue endpoints with direct measurement of aerobic capacity. Group A had a higher dropout rate (24%) compared to groups B (17%) and C (17%). Peak oxygen consumption (VO2 peak) increased by 43% (p < 0.05), anaerobic threshold (VO2AT) by 37% (p < 0.05) and exercise time by 33% (p < 0.05) after training in Group A; by 24% (p < 0.05), 18% (p < 0.05) and 22% (p < 0.05), respectively, in B; and by 17% (p < 0.05), 8% (p < 0.05) and 14% (p < 0.05), respectively, in C; while both remained almost unchanged in Group D. These results demonstrate that intense exercise training on non-dialysis days is the most effective way of training, whereas exercise during HD is also effective and preferable. PMID- 11900262 TI - Improved client participation in the rehabilitation process using a client centred goal formulation structure. AB - The aim was to evaluate whether the use of a client-centred instrument, the Canadian Occupational Performance Measure (COPM), affects the patients' perception of active participation in the rehabilitation process. The study included 155 patients in the experiment group and 55 in the control group, within geriatric, stroke, and home rehabilitation. The COPM was used in the experiment group. A structured interview was performed within 2-4 weeks after discharge with 88 patients in the experiment group and 30 patients in the control group. The results show significant differences between the groups. More patients in the experiment group perceived that treatment goals were identified, were able to recall the goals, felt that they were active participants in the goal formulation process, and perceived themselves better able to manage after completed rehabilitation compared with patients in the control group. The study indicates that the COPM improves client participation in the rehabilitation process. PMID- 11900263 TI - Multiplex PCR-based detection and identification of the most common Salmonella second-phase flagellar antigens. AB - Most Salmonella serotypes alternatively express phase 1 or phase 2 flagellar antigens encoded by fliC and fljB genes respectively. Flagellar phase reversal to identify both flagellar antigens is not necessary at the genetic level. Variable internal regions of the fljB genes encoding H:1,w, H:e,n,x and H:e,n,z15 antigens have been sequenced and the specific sites for each antigen determined in selected Salmonella serotypes. These results, together with flagellar H1 complex variable internal sequences previously published, have been used to design a multiplex-PCR to identify H:1,2, H:1,5, H:1,6, H:1,7, H:1,w, H:e,n,x and H:e,n,z15 second-phase antigens. These antigens are part of the most common Salmonella serotypes possessing second-phase flagellar antigens. This multiplex PCR includes 10 primers. A total of 140 Salmonella strains associated with 49 different serotypes were tested. Each strain generated one second-phase-specific antigen fragment, ranging between 50 and 400 bps. Twenty-five strains associated with 17 serotypes, with no second-phase antigen or with an antigen different from those tested in this work, did not generate any fragments. The method is quick, specific and reproducible and is independent of the phase expressed by the bacteria when tested. PMID- 11900264 TI - Erratum to "Lactobacillus sakei: recent developments and future prospects" [Research in Microbiology 152 (2001) 839]. AB - Lactobacillus sakei is one of the most important bacterial species involved in meat preservation and meat fermentation. In the last fifteen years, numerous studies have focused on this species due to its important role in food microbiology. The present paper reviews current knowledge of this emerging species in the fields of taxonomy, phylogeny and physiology, and metabolism. Recent developments in genetic tools and molecular genetics will also be emphasized to evaluate future prospects. PMID- 11900266 TI - In vitro adhesive properties and virulence factors of Enterococcusfaecalis strains. AB - Twenty-nine Enterococcus faecalis isolates from patients with endocarditis or bacteremia or from stools of healthy volunteers were investigated for their ability to adhere to Int-407 and Girardi heart cell lines and for the presence of known enterococcal virulence factors. Eight strains (27.6%) adhered predominantly to Int-407 cells. The adherence of enterococci was enhanced by proteolytic digestion, suggesting that some cell binding components become surface-exposed after treatment with trypsin. The occurrence of known potential virulence factors of enterococci among these strains was determined and was as follows: enterococcal surface protein (72.4%), gelatinase (58.6%), aggregation substance (48.3%) and cytolysin (17.2%). Bacterial adherence was not significantly associated with any of these virulence factors. PMID- 11900265 TI - Cloning and characterization of two catechol 1,2-dioxygenase genes from Acinetobacter radioresistens S13. AB - Two novel catechol 1,2-dioxygenase (C 1,2-O) genes have been isolated from an Acinetobacter radioresistens strain that grows on phenol or benzoate as sole carbon and energy source. Designated as catA(A) and catA(B), they encode proteins composed of 314 and 306 amino acids, whose deduced sequences indicate that they have approximately 53% identity, whereas their NH2-terminal and COOH-terminal regions have no sequences in common. This may explain their different thermal and pH stability. Polyclonal antibodies raised against an amino-terminal CatA(A) peptide or the whole CatA(B) protein were used to establish their inducible and differential expression patterns upon bacterial growth in phenol or benzoate. The CatA(A) protein (IsoA) was induced by both phenol and benzoate though with different kinetics, whereas the catA(B) product (IsoB) was constitutively produced at low levels that increased only during growth in the presence of benzoate. PMID- 11900267 TI - Spoligotyping and IS6110-RFLP typing of Mycobacterium tuberculosis from French Guiana: a comparison of results with international databases underlines interregional transmission from neighboring countries. AB - In this investigation, 94 clinical isolates of Mycobacterium tuberculosis obtained over a 3-year period (1996-1998) from French Guiana were characterized by spoligotyping and IS6110-RFLP and the patterns obtained were compared with genotypes representing the worldwide diversity in an international spoligotyping database (n = 4269) and a IS6110-RFLP database (n = 4189). All the clustered isolates giving < or = 6 copies of IS6110 were further typed using the double repetitive element (DRE)-PCR. The results obtained underlined the highly diverse nature of the M. tuberculosis population in French Guiana with potential links to neighboring countries within the Americas. It may be hypothesized that the genetic heterogeneity of tubercle bacilli in French Guiana is linked to the high number of imported cases of tuberculosis, that may account for as high as 68% of all tuberculosis cases. Although an epidemiological investigation based on direct interrogation of patients was not performed, available medical records suggested that the clustering of isolates was mostly linked to the following risk factors: pulmonary tuberculosis, smear-positive samples, foreign-born nationals and/or immigrants, and a high rate of HIV-TB coinfection. Thus the persisting foci of endemic disease and increased active transmission due to high population flux and HIV coinfection may be largely responsible for the relatively high incidence of tuberculosis in French Guiana. PMID- 11900268 TI - Occurrence and properties of glutathione S-transferases in phenol-degrading Pseudomonas strains. AB - Pseudomonas sp. strains, able to degrade aromatic compounds such as phenol, were chosen to investigate the occurrence and characteristics of glutathione S transferases (GSTs). Affinity chromatography purification showed the presence of at least one GST in each studied strain. The purified proteins exhibited a great variety in the N-terminal sequences and different enzyme activities with the standard GST substrates tested. Two Pseudomonas strains, M1 and CF600, were chosen to investigate the GST activities under different growth conditions. Therefore, cells were grown either on phenol or on different nonaromatic carbon sources in the presence and absence of increasing phenol concentrations. In strain M1 a strong correlation between the activities of the catechol 1,2 dioxygenase and GST was observed in all the tested conditions. Moreover, growth on different organic acids also affected GST activity levels, with a negative correlation with the specific growth rate determined by each substrate. These results suggest a possible function of GST as a response to specific metabolic conditions determined by phenol toxicity and/or catabolism and the metabolic status of the cells. The same experiments performed with the CF600 strain did not show induction of GST activity in any of the tested conditions, indicating that GST_CF600 probably has a different role in cell metabolism. Native gel electrophoresis gave indications that GST dimerization could be an important process in the modulation of GST activity. PMID- 11900269 TI - Effects of tibolone on endogenous nitric oxide (nitrite/nitrate levels) in postmenopausal women. AB - OBJECTIVE: To observe the effects of hormone replacement therapy with 2.5 mg/day tibolone for 6 months upon plasma nitrite/nitrate (end products of nitric oxide metabolism) in postmenopausal women. PATIENTS AND METHODS: The study involved 24 healthy postmenopausal women treated with tibolone (2.5 mg/day for 6 months) and 20 postmenopausal women who received placebo. Plasma nitrite/nitrate levels were measured with the Griess reaction. Statistical significance was analyzed by the Mann-Whitney U test and Pearson correlation analysis. RESULTS: The mean baseline concentrations of nitrite/nitrate were similar in both treated and placebo groups. Tibolone treatment decreased plasma nitrite/nitrate levels. CONCLUSIONS: We suggest that because of its androgenic and progestogenic actions, tibolone diminishes nitrite/nitrate levels. Also our data provide evidence for the existence of increased (1/3 of subjects, n = 8) and reduced (2/3 of subjects, n = 16) nitrite/nitrate levels in response to tibolone treatment. Clearly, further studies are necessary to establish a risk/benefit ratio for tibolone in view of its effects on nitric oxide metabolism. PMID- 11900270 TI - Female sexual dysfunction. AB - Happiness is an attitude, not an event! The glass is either half-full or half empty; it simply depends on the viewer. Some rich people are miserable...some poor people are ecstatic. You cannot change the world but you can change yourself. You, and you alone, control your destiny. With very little practice, you can learn to think positively. Postmenopausal estrogen deficiency leads to urogenital atrophy. Sexual dysfunction and urinary dysfunction are the most inevitable but least publicized consequences of estrogen deficiency, and these represent important quality-of-life issues that patients and health care providers are often reluctant to discuss. In addition to estrogen deficiency, oophorectomized women may be subject to androgen deficiency and problems with libido. While the relationship with one's partner is the quintessential factor in female sexuality, hormone deficiency remains important, especially in reference to genital atrophy. Humans are the products of learned behavior. We literally become what we think. "I am sick and tired" becomes a self-fulfilling prophecy. Self-esteem represents learned behavior just as ego does...but there is a world of difference between the two. Sexuality is enhanced by good self-esteem and hindered by ego: two partners working together can reach far greater heights than either alone ever imagined...one and one can equal three! There are good data to substantiate that happy people have more sex and that people who have more sex are happier. Partners need to learn to honor one another, communicate in a positive manner, develop touching salutations, be a spouse at home, and make their bedroom a sacred sanctuary. With a little behavior modification and hormone therapy, sexuality can remain a priority ad infinitum. PMID- 11900271 TI - Osteoporosis in postmenopausal women (Mexico City): 1. Risk factors. AB - OBJECTIVE: To determine risk factors for osteoporosis in postmenopausal women from Mexico City. METHOD: The following prognostic variables for low bone density were analyzed in 400 postmenopausal women: age, body mass index (BMI), time since menopause (TM), previous and current hormone replacement therapy (HRT), previous use of corticosteroids, thyroid disease, and previous fractures. All patients were submitted to bone absorptiometry (DEXA) considering the diagnostic criteria for osteoporosis established by the WHO. All variables were analyzed in a bivariate mode, and logistic regression analysis was performed for variables that proved significant (age, TM, and BMI), estimating odds ratios and 95% confidence intervals. RESULTS: Age, TM and BMI resulted as significant risk factors for osteoporosis (P < .01), with the following cut-off points: age < or = 48 and >48 years; TM < or = 5 and >5 years; and BMI < or = 32 and >32 for L1-L4 average, BMI < or = 30 and >30 for femoral neck (FN), < or = 27 and >27 for trochanter, and < or = 32 and >32 for Ward's triangle (WT). Odds ratio (OR) for age was 1.57 at L1 L4 average (95% CI 1.01-2.47); OR for TM was 2.09 at L1-L4 average (95% CI 1.24 3.55), 2.05 at FN (95% CI 1.23-3.47) and 1.78 in WT (95% CI 1.05-2.99). OR for BMI was 0.36 at L1-L4 average (95% CI 0.17-0.80), 0.25 at FN (95% CI 0.12-0.5), 0.18 at trochanter (95% CI 0.10-0.32) and 0.29 at WT (95% CI 0.13-0.63). CONCLUSIONS: The various risk factors exerted different influences on the bone areas analyzed. Age increased the risk for lumbar spine osteoporosis, TM had a deleterious effect, while BMI showed a beneficial effect on bone density. These factors should be considered to establish the risk of osteoporosis in postmenopausal women, and to decide which patients should be submitted to absorptiometry. PMID- 11900272 TI - Osteoporosis in postmenopausal women (Mexico City): 2. Validation of a predictive clinical index. AB - OBJECTIVE: To analyze and validate a clinical index (CI) in postmenopausal women that will help to determine the need for bone absorptiometry. METHODS: 1088 postmenopausal women were studied; age, body mass index (BMI) and time since menopause (TM) were analyzed. A score was attributed to each variable (0 points if it did not increase the risk of osteoporosis or 1 point if it increased it): age < or = 48 years = 0 points, >48 years = 1 point; BMI < or = 32 = 1 point or >32 = 0 points for lumbar spine and Ward's triangle (WT); BMI < or = 30 = 1 point or >30 = 0 points for femoral neck (FN); TM < or = 5 years = 0 points or >5 years = 1 point. In all cases, bone absorptiometry (DEXA) was done. Comparison between groups was determined by Student's t test. Regression models were used to establish possible associations between bone absorptiometry and the clinical variables studied. Univariate and multivariate analyses were performed. RESULTS: The most highly significant differences were observed starting from 2 points in lumbar spine and FN. The most important difference in WT was detected starting from 1 point. In L1-L4 average, a significant correlation was established between the CI and age, TM, and BMI (r = 0.707; P < .001), as well as between the CI and the T score (r = 0.302; P < .001). FN analysis showed a correlation between the CI, the TM, and BMI (r = 0.720; P < .001) and between the CI and T score (r = 0.338; P < .001). WT analysis revealed a correlation between the CI and TM, and BMI (r = 0.698; P < .001) and between the CI and T score (r = 0.324; P < .001). CONCLUSION: It is reasonable to conclude that a score of 2 points in the lumbar spine and FN, and 1 point in WT, suggests the need for bone absorptiometry. PMID- 11900273 TI - Sexual dysfunction: male and female issues. AB - In March 1998, a new sexual revolution began. It was at that time that Viagra, a pill for the treatment of male erectile dysfunction, was introduced. Release of this medication not only allowed scientists to identify the basic physiology of erectile functioning but also for the first time allowed men to reverse the natural aging process of loss of an erection. This, unfortunately, failed to address the other most important part of sexual relations: the female partner. It is not until recently that we have begun to recognize the vast importance of the couple's relationship. This paper reviews the pathophysiology of both male and female sexual dysfunctions and their relationships. It describes the testing process that is involved in the ideology and management of sexual dysfunction. The treatment options reviewed include the use of the older methods for male sexual dysfunction, as well as the new pharmacologic treatments and their potential. The treatment of female sexual dysfunction is also addressed. PMID- 11900274 TI - Does hormonal replacement therapy actually protect against cardiovascular disease? AB - It is generally accepted that menopausal symptoms can be controlled by HRT. As additional (putative) benefits of HRT were discovered, it was claimed that such uses were also important. Soon, trials were instituted to test these assertions, some of them quite large and projected to last a long time (> or = 5 years). The most instructive for medical practice are the observational trials and, still more, randomized, controlled trials. Unfortunately, the results of these trials for both primary and secondary prevention of CHD are not in agreement, although the observational trials are mostly more favorable to the benefits of HRT on CHD. The present review considers both kinds of trials and their effects on primary and secondary CHD prevention. It also attempts to group the studies by their results and their implications for guiding patients in choices and decisions regarding HRT. PMID- 11900275 TI - Markers for risk of type 1 diabetes in relatives of Alsacian patients with type 1 diabetes. AB - BACKGROUND: The cytotoxic T lymphocyte-associated antigen 4 gene (CTLA-4) encode the T cell receptor involved in the control of T cell proliferation and mediates T cell apoptosis. The receptor protein is a specific T lymphocyte surface antigen that is detected on cells only after antigen presentation. Thus, CTLA-4 is directly involved in both immune and autoimmune responses and may be involved in the pathogenesis of multiple T cell-mediated autoimmune disorders. There is polymorphism at position 49 in exon 1 of the CTLA-4 gene, providing an A-G exchange. Moreover, we assessed the CTLA-4 49 (Thr/Ala) polymorphism in diabetic patients and first-degree relatives as compared to control subjects. RESEARCH DESIGN AND METHODS: Three loci (HLA-DQB1, DQA1 and CTLA-4) were analysed in 62 type 1 diabetic patients, 72 first-degree relatives and 84 nondiabetic control subjects by means of PCR-RFLP. RESULTS: A significant enrichment in DQB1 alleles encoding for an amino acid different from Asp in position 57 (NA) and DQA1 alleles encoding for Arg in position 52 was observed in diabetic subjects and first-degree relatives as compared to controls. The genotype and allele frequencies of these polymorphisms in type 1 diabetic patients and first-degree relatives differed significantly from those of controls (p < 0.001 and 0.05 respectively). CTLA-49 Ala alleles frequencies were 75.8% in type 1 diabetic patients and 68.1% in first-degree relatives in comparison to 35.7% in control subjects. The Ala/Ala genotype conferred a relative risk of 18.8 (p < 0.001). CONCLUSION: The CTLA-4 49 Ala allele confers an increased risk of type 1 diabetes, independent of age and HLA-DQ genetic markers. PMID- 11900276 TI - Effect of aldose reductase inhibition on interleukin-1beta-induced nitric oxide (NO) synthesis in vascular tissue. AB - Glucose metabolism via sorbitol pathway has been implicated as a possible contributor to the diabetes-related vascular changes. Nitric oxide plays a major regulatory role in the vascular dilatatory and constricted response. Also it has been observed that diabetes causes vascular changes leading to a decrease in nitric oxide production. Additionally the accumulation of sorbitol is also related to decreased nitric oxide production. In the present study we investigated the effect of normal and high glucose in the presence or absence of both interleukin-1beta or an aldose reductase inhibitor on nitric oxide production in rat aortic rings in vitro. Aortic rings from normal male Wistar rats were dissected and incubated for 24 to 48 hrs in the presence of glucose (5.0 mM or 20 mM) or with or without interleukin (20 ng/ml). Other rings were incubated in the above media with the addition of the aldose reductase inhibitor (WAY 121509). Interleukin-1beta stimulated the 24 hr nitric oxide production and WAY 121509 decreased it under both low and high glucose culture conditions. The interleukin-1beta stimulation was continued for 72 hrs. Nitric oxide production in response to interleukin-1beta was greater at all time points when compared to the incubation in media without interleukin-1beta. In media containing WAY 121509 the nitric oxide production was decreased. Interleukin-1beta stimulated a greater increase in nitric oxide production from aortic rings when incubated in high glucose when compared to normal glucose. The inhibitory effect of aldose reductase inhibition was reversible after 24 hr inhibition under both normal and high glucose conditions. We conclude that high glucose enhances the interleukin 1beta-induced nitric oxide synthesis and the cytokine-induced nitric oxide production was inhibited by aldose reductase inhibition. Nitric oxide production may be linked to redox influences caused by the polyol pathway. PMID- 11900278 TI - The non-immune RIP-Kb mouse is a useful host for islet transplantation, as the diabetes is spontaneous, mild and predictable. AB - Chemically-induced diabetic mice and spontaneously diabetic NOD mice have been valuable as recipients for experimental islet transplantation. However, their maintenance often requires parenteral insulin. Diabetogenic chemicals can be cytotoxic to the host's immune system and to other organs some of which are often used as the transplant site. Procurement of diabetic cohorts in the NOD mouse is problematic due to variability in the age of disease onset. We show that RIP-Kb mice, which spontaneously develop non-immune diabetes due to over-expression of the H-2Kb heavy chain in beta cells, offer many advantages as islet transplant recipients. Diabetes is predictable with a relatively narrow range of onset (4 wk) and blood glucose levels (23.0 +/- 4.0 mmol/l for 39 males at 6 weeks of age). The diabetes is mild enough so that most diabetic mice can be maintained to 40 weeks of age without parenteral insulin. This consistency of diabetes avails that outcomes of intervention can be interpreted with confidence. PMID- 11900277 TI - Effect of treating streptozotocin-induced diabetic rats with sorbinil, myo inositol or aminoguanidine on endoneurial blood flow, motor nerve conduction velocity and vascular function of epineurial arterioles of the sciatic nerve. AB - Previously we have demonstrated that diabetes causes impairment in vascular function of epineurial vessels, which precedes the slowing of motor nerve conduction velocity. Treatment of diabetic rats with aldose reductase inhibitors, aminoguanidine or myo-inositol supplementation have been shown to improve motor nerve conduction velocity and/or decreased endoneurial blood flow. However, the effect these treatments have on vascular reactivity of epineurial vessels of the sciatic nerve is unknown. In these studies we examined the effect of treating streptozotocin-induced rats with sorbinil, aminoguanidine or myo-inositol on motor nerve conduction velocity, endoneurial blood flow and endothelium-dependent vascular relaxation of arterioles that provide circulation to the region of the sciatic nerve. Treating diabetic rats with sorbinil, aminoguanidine or myo inositol improved the reduction of endoneurial blood flow and motor nerve conduction velocity. However, only sorbinil treatment significantly improved the diabetes-induced impairment of acetylcholine-mediated vasodilation of epineurial vessels of the sciatic nerve. All three treatments were efficacious in preventing the appropriate metabolic derangements associated with either activation of the polyol pathway or increased nonenzymatic glycation. In addition, sorbinil was shown to prevent the diabetes-induced decrease in lens glutathione level. However, other markers of oxidative stress were not vividly improved by these treatments. These studies suggest that sorbinil treatment may be more effective in preventing neural dysfunction in diabetes than either aminoguanidine or myo inositol. PMID- 11900279 TI - D-chiro-inositol--its functional role in insulin action and its deficit in insulin resistance. AB - In this review we discuss the biological significance of D-chiro-inositol, originally discovered as a component of a putative mediator of intracellular insulin action, where as a putative mediator, it accelerates the dephosphorylation of glycogen synthase and pyruvate dehydrogenase, rate limiting enzymes of non-oxidative and oxidative glucose disposal. Early studies demonstrated a linear relationship between its decreased urinary excretion and the degree of insulin resistance present. When tissue contents, including muscle, of type 2 diabetic subjects were assayed, they demonstrated a more general body deficiency. Administration of D-chiro-inositol to diabetic rats, Rhesus monkeys and now to humans accelerated glucose disposal and sensitized insulin action. A defect in vivo in the epimerization of myo-inositol to chiro-inositol in insulin sensitive tissues of the GK type 2 diabetic rat has been elucidated. Thus, administered D-chiro-inositol may act to bypass a defective normal epimerization of myo-inositol to D-chiro-inositol associated with insulin resistance and act to at least partially restore insulin sensitivity and glucose disposal. PMID- 11900280 TI - Streptozotocin-induced diabetes decreases mammary gland lipoprotein lipase activity and messenger ribonucleic acid in pregnant and nonpregnant rats. AB - Diabetes mellitus is associated with a reduction of lipoprotein lipase (LPL) activity in adipose tissue and development of hypertriglyceridemia. To determine how a condition of severe insulin deficiency affects mammary gland LPL activity and mRNA expression during late pregnancy, streptozotocin (STZ) treated (40 mg/kg) and non-treated (control) virgin and 20 day pregnant rats were studied. In control rats, both LPL activity and mRNA were higher in pregnant than in virgin rats. When compared to control rats, STZ-treated rats, either pregnant or virgin, showed decreased LPL activity and mRNA content. Furthermore, mammary gland LPL activity was linearly correlated with mRNA content, and either variable was linearly correlated with plasma insulin levels. Thus, insulin deficiency impairs the expression of LPL in mammary glands, revealing the role of insulin as a modulator of the enzyme at the mRNA expression level. PMID- 11900281 TI - Growth factor-dependent proliferation of the pancreatic beta-cell line betaTC tet: an assay for beta-cell mitogenic factors. AB - The ability to expand normal pancreatic islet beta cells in culture would significantly advance the prospects of cell therapy for diabetes. A number of growth factors can stimulate limited islet cell replication, however other factors may exist which are more effective beta-cell-specific mitogens. The search for novel beta-cell growth factors has been hampered by the lack of a beta cell-specific proliferation assay. We developed a simple and sensitive assay for beta-cell growth factors based on a conditionally-transformed mouse beta-cell line (betaTC-tet). These cells express the SV40 T antigen (Tag) oncoprotein under control of the tetracycline (Tc) operon regulatory system. In the presence of Tc, Tag expression is tightly shut off and the cells undergo complete growth arrest. Here we show that the growth-arrested cells can proliferate in response to growth factors in the absence of Tag. Using this assay, a number of growth factors previously shown to be mitogenic to a mixed islet cell population were found to induce proliferation of pure beta cells. We conclude that growth-arrested betaTC tet cells can be employed in a survey of factors from various sources for identifying novel factors with beta-cell mitogenic activity. PMID- 11900282 TI - Pulmonary vein paced activation sequence mapping: comparison with activation sequences during onset of focal atrial fibrillation. AB - INTRODUCTION: Atrial fibrillation (AF) may originate from a single focus, with the vast majority observed within the pulmonary veins. To facilitate mapping, we hypothesized that there would be a characteristic right atrial endocardial activation sequence pattern associated with pacing and spontaneous focal activity from each of the four pulmonary veins. METHODS AND RESULTS: In 10 patients with focal AF, a standardized set of catheters was positioned in the right atrium. These included a 20-pole catheter along the crista terminalis, a decapolar catheter in the coronary sinus (CS), and a His-bundle electrode. Pacing (700 and 300 msec) was performed with a mapping catheter from each of the four pulmonary veins. Activation sequence maps were created by measurement of activation times to each of the recording bipoles with the proximal CS bipole as the arbitrary reference point. Similar maps were constructed for the activation sequence of the pulmonary vein ectopic that initiated AF. There was a characteristic right atrial activation map created by pacing each pulmonary vein that corresponded closely with the map from the same pulmonary vein during initiation of focal AF. The pulmonary vein of origin could be distinguished on the basis of this characteristic pattern and some stereotypic observations. CS activation occurred proximal to distal for right pulmonary veins and distal to proximal for left pulmonary veins. Significant differences in activation timing between the CS and crista terminalis differentiated upper from lower pulmonary veins. CONCLUSION: There is a characteristic right atrial activation map for activity arising from each of the four pulmonary veins that corresponded closely with the map from the same pulmonary vein during initiation of focal AF. These findings may facilitate mapping and ablation of focal AF. PMID- 11900283 TI - To map or not to map. PMID- 11900284 TI - Differentiation of atrial and pulmonary vein potentials recorded circumferentially within pulmonary veins. AB - INTRODUCTION: Accurate discrimination of atrial and pulmonary vein potentials recorded circumferentially within the pulmonary veins is important when performing segmental isolation of the pulmonary veins in patients with atrial fibrillation. METHODS AND RESULTS: Twenty patients with paroxysmal atrial fibrillation underwent pulmonary vein mapping with a decapolar Lasso catheter during sinus rhythm and during pacing in the distal coronary sinus and left atrial appendage. Bipolar and unipolar electrograms were recorded within the left superior, right superior, and left inferior pulmonary veins. The atrial potentials were larger in the left pulmonary veins than in the right superior pulmonary vein, whereas the pulmonary vein potentials in the superior pulmonary veins were larger than in the left inferior pulmonary vein. The atrial and pulmonary vein potentials usually were readily distinguished during sinus rhythm in the right superior pulmonary vein. Characteristic distribution and morphologies of the atrial potentials as well as the response to distal coronary sinus and left atrial appendage pacing were useful for differentiating the atrial and pulmonary vein potentials in the left pulmonary veins. CONCLUSION: Atrial and pulmonary vein potentials recorded circumferentially within the pulmonary veins have characteristic features that are useful in distinguishing them from each other. In the left pulmonary veins, discrimination of the atrial and pulmonary vein potentials is aided by coronary sinus or left atrial appendage pacing. PMID- 11900285 TI - Paroxysmal cycle length shortening in the pulmonary veins during atrial fibrillation correlates with arrhythmogenic triggering foci in sinus rhythm. AB - INTRODUCTION: The focal origin of atrial fibrillation (AF) is identified by recording atrial ectopic beats or the ectopic activity that precedes AF. We hypothesized that arrhythmogenic pulmonary veins (PVs) also could be identified during persistent AF. METHODS AND RESULTS: Patients with persistent AF referred for focal ablation were enrolled prospectively. During AF, bipolar electrograms were recorded from each PV for a minimum of 120 seconds, as well as from the right atrium and coronary sinus. The cycle length of activity in each PV was measured during AF and plotted on a frequency histogram. Following cardioversion to sinus rhythm, arrhythmogenic PVs were identified from reinitiation of AF or from ectopic beats. Ten patients were enrolled and 37 PVs analyzed. During AF, 17 PVs demonstrated bimodal cycle length frequency histograms, with periods of paroxysmal short cycle length recording. Following cardioversion, 14 PVs were identified as arrhythmogenic as defined earlier. Each of these arrhythmogenic PVs showed paroxysmal short cycle length recording during AF. Sensitivity was 87%, specificity 91%, positive predictive value 87%, and negative predictive value 100%. CONCLUSION: The arrhythmogenic PVs responsible for the focal activity that triggers AF also demonstrate paroxysmal short cycle length recording during sustained AF. These results demonstrate that arrhythmogenic PVs still can be identified reliably, even during sustained AF. PMID- 11900286 TI - Finish what you started? PMID- 11900287 TI - Sustained monomorphic ventricular tachycardia ablation from the aortic sinus of valsalva. AB - INTRODUCTION: The aim of this study was to delineate the electrophysiologic mechanisms of a novel type of ventricular tachycardia (VT) originating from the aortic sinus of Valsalva. METHODS AND RESULTS: Endocardial mapping was performed in four patients with symptomatic VT originating from the aortic sinus of Valsalva. Two patients suffered from dilative cardiomyopathy; the other two patients had no structural heart disease. Five VTs could be induced and terminated by programmed ventricular stimulation. Successful ablation was performed in the noncoronary sinus of Valsalva in three VTs and in the left aortic sinus in two. Abnormal (diastolic or presystolic) potentials were recorded during sinus rhythm (mean interval from the end of QRS complex to the potential 121+/-98 msec) and during VT (mean interval from the potential to QRS complex 64+/-45 msec) at effective sites in the aortic sinuses of Valsalva. Concealed entrainment was demonstrated at all successful sites. VT recurred in one patient after 1 month, whereas no recurrences were observed during follow-up of 8+/-6 months in the other three patients. CONCLUSION: Reentry constitutes one mechanism of VT originating from the aortic sinus of Valsalva. Entrainment mapping is useful to characterize the reentrant circuit of these VTs and to guide ablation. PMID- 11900288 TI - Does a policy of repeated early cardioversion for recurrence of atrial fibrillation work? PMID- 11900290 TI - Effects of verapamil and ibutilide on atrial fibrillation and postfibrillation atrial refractoriness. AB - INTRODUCTION: Early recurrence of atrial fibrillation (AF) after cardioversion may be related to shortening of the atrial effective refractory period (ERP). This study compared the effects of verapamil and ibutilide on AF cycle length (AFCL), atrial ERP, and susceptibility to recurrent AF. METHODS AND RESULTS: In 33 adults, the atrial ERP was measured at basic drive CLs of 350 and 500 msec before and after a brief episode of pacing-induced AF. During AF, verapamil, ibutilide, or saline was infused in 11 patients each. Shortening of the post-AF atrial ERP was attenuated by verapamil and prevented by ibutilide. AFCL shortened by 32+/-21 msec in the verapamil group (P < 0.01), prolonged by 44+/-14 msec in the ibutilide group (P < 0.001), and did not change in the control group. AF converted to sinus rhythm within 10 minutes less often after verapamil (0%) than after ibutilide (82%) or than in the control group (73%). Post-AF, AF lasting > 10 minutes was induced more often in the verapamil group than in the ibutilide group (26% vs 0%; P = 0.01). Another 10 patients received verapamil or ibutilide in the absence of AF. Atrial ERP was unchanged after verapamil and prolonged after ibutilide. CONCLUSION: Verapamil shortens AFCL and impedes the conversion of induced AF, whereas ibutilide prolongs AFCL and does not impede the early conversion of induced AF. Ibutilide is more effective than verapamil in preventing pos PMID- 11900289 TI - Analysis of troponin I levels after spontaneous implantable cardioverter defibrillator shocks. AB - INTRODUCTION: Serum cardiac troponin I (cTnI) is a sensitive and specific marker for myocardial injury. Myocardial ischemia and/or injury can be a trigger for ventricular arrhythmias. The aim of this study was to assess the frequency and significance of elevated serum cTnI levels after spontaneous implantable cardioverter defibrillator (ICD) shocks. METHODS AND RESULTS: Serial cTnI measurements and ECGs were performed in 35 patients with transvenous ICDs who were admitted after spontaneous ICD shocks. Elevated cTnI levels were found in 18 patients (51%). Acute coronary syndrome was diagnosed in 5 (22%) of 23 patients with known coronary artery disease. After excluding the patients with acute coronary syndrome, elevated cTnI levels were present in 13 (43%) of 30 patients: 18% of patients with < or =3 shocks and 58% of patients with >3 shocks. Patients with elevated cTnI levels received a higher number of shocks (16+/-18 vs 5+/-7; P < 0.05) and had higher total delivered energies (475+/-538 J vs 128+/-184 J; P < 0.05) compared with patients with normal cTnI. Patients with acute coronary syndrome had higher peak cTnI levels (18+/-16 ng/mL) compared with patients with elevated cTnI without acute coronary syndrome (3.8+/-4.3 ng/mL; P < 0.01). CONCLUSION: Serum cTnI rises occur in the majority of patients after multiple (>3) spontaneous ICD discharges but are due to acute coronary syndrome only 14% of the time (22% of the time in patients with known coronary artery disease). PMID- 11900291 TI - Atrial morphology in hearts with congenitally corrected transposition of the great arteries: implications for the interventionist. AB - INTRODUCTION: In view of the possible need for septal puncture to ablate left sided lesions and the occasional difficulty in coronary sinus (CS) cannulation, we investigated relevant anatomic features in the right atrium of hearts with congenitally corrected transposition of the great arteries (ccTGA). METHODS AND RESULTS: Nine hearts with ccTGA and an intact atrial septum and eight weight matched normal hearts were examined by studying the "septal" aspect of the right atrium with reference to the oval fossa (OF). The anterior margin was arbitrarily measured as the shortest distance from the OF to the superior mitral/tricuspid annulus. The posterior margin was measured from the OF to the posterior-most edge of the right atrial "septal" surface. The total "septal" surface width was measured at the middle of the OF. The stretched OF dimensions and CS isthmus length were noted. Mann-Whitney test was used to compare absolute and indexed dimensions, i.e.. normalized to total width. The posterior margin in hearts with ccTGA was shorter than in controls (6.3+/-2.4 mm vs 11+/-1.9 mm, P < 0.001; normalized margin P = 0.09). The CS isthmus also was significantly shorter (5.3+/ 2.7 mm vs 11.4+/-2.2 mm, P < 0.001). In two hearts with ccTGA, the CS opening into the right atrium was on the same side of the eustachian valve as the inferior caval vein. CONCLUSION: The shorter posterior "septal" margin in hearts with ccTGA may increase the risk of exiting the heart while performing septal puncture when pointing the needle posteriorly. The shorter CS isthmus and the abnormal location of the CS opening in some of these hearts are important when contemplating radiofrequency ablation in this area. PMID- 11900292 TI - Importance of right and left atrial dilation and linear ablation for perpetuation of sustained atrial fibrillation. AB - INTRODUCTION: Atrial dilation associated with increasing atrial pressure plays an apparent role in the development of atrial fibrillation (AF). We characterized a new model of separate and biatrial dilation in the Langendorff-perfused rabbit heart. The aim of this study was to examine if sustained AF in this model (1) would be inducible by separate right atrial (RA) and left atrial (LA) dilation; (2) would be reproducibly inducible at the same pressure level; and (3) could be suppressed by RA, LA, or biatrial ablation. METHODS AND RESULTS: Intra-atrial pressure was increased stepwise in the RA (n = 13), LA (n = 12), or both atria (n = 25) until sustained AF could be induced or a pressure of 20 cm H2O was reached. The stimulation protocol was repeated once in RA and LA dilation (n = 9) and three times in biatrial dilation (n = 7). Then, RA orifices (superior and inferior caval veins, tricuspid valve annulus, and foramen ovale) or LA orifices (pulmonary veins, mitral valve annulus, and foramen ovale) were connected by radiofrequency (RF) lesions. Sustained AF was rendered inducible in 100% of hearts with biatrial dilation, but in only 92% of hearts with RA dilation and 67% with LA dilation. Inducibility of sustained AF was reproducible. Under biatrial dilation, not RA ablation (0/10 hearts; P = NS) but LA ablation (4/11 hearts; P < 0.05) and biatrial ablation (16/21; P < 0.01) reduced the inducibility of sustained AF. CONCLUSION: The inducibility of sustained AF due to increased intra atrial pressure differs between the RA and LA. LA and biatrial lesions, not RA RF lesions, reduce the ability to perpetuate sustained AF. PMID- 11900294 TI - Reexpression of the nifedipine-resistant calcium channel during dedifferentiation of adult rat ventricular cardiomyocytes. AB - INTRODUCTION: Using an adult rat ventricular culture model and whole-cell patch clamp technique, we investigated whether the nifedipine-resistant calcium current observed at the neonatal stage but not at the adult stage could be reobserved under dedifferentiating conditions. METHODS AND RESULTS: Application of 2 microM nifedipine totally inhibited the inward calcium current (carried by 5 mM Ba2+) in freshly isolated cells from adult rat heart, but it failed to block it completely when cells are cultured for 8 to 12 days. Dose-response curves of nifedipine in the range from 2 to 50 microM showed a residual current that represented, in the presence of 2 microM nifedipine, 16.4%+/-1.8% (n = 10) and 20.4%+/-1.5% (n = 10) of the total current in 8- and 12-day-old cultured cells, respectively. In these conditions, its density at 0 mV increased slightly during culture (-2.1+/-0.2 pA/pF, n = 7, and -3.2+/-0.4 pA/pF, n = 8, after 8 and 12 days in culture, respectively), without modification in the current-to-voltage curve, as well as in kinetics properties. CONCLUSION: These results show that nifedipine-resistant calcium current, which has been shown to be developmentally regulated in rat ventricular cardiomyocytes, can be reexpressed in cultured-dedifferentiated cells. Its possible expression and implication in physiopathologic function are suggested. PMID- 11900293 TI - Prominent I(Ks) in epicardium and endocardium contributes to development of transmural dispersion of repolarization but protects against development of early afterdepolarizations. AB - INTRODUCTION: Previous studies from our laboratory demonstrated (1) a much larger I(Ks) and (2) inability to induce early afterdepolarization (EAD) activity in epicardial and endocardial cells versus M cells. This study tests the hypothesis that these two characteristics are interrelated. METHODS AND RESULTS: Standard and floating microelectrode techniques were used to record transmembrane activity from the canine left ventricular epicardial, M, and endocardial regions in isolated tissue slices and arterially perfused wedge preparations. The I(Kr) blocker E-4031 (1 to 10 microM) caused prominent prolongation of action potential duration (APD) and induced EADs in tissues isolated from the M region, but not those from epicardium or endocardium, causing a large transmural dispersion of APD. In contrast, the I(Ks) blocker chromanol 293B (10 to 30 microM) produced moderate prolongation of APD without EADs in all three tissue types. The combination of E-4031 (1 microM) and chromanol 293B (30 microM) resulted in profound prolongation of APD and the development of EADs in all three tissue types. In the perfused wedge, neither E-4031 nor chromanol 293B alone could induce EADs. In combination, the two drugs caused significant prolongation of APD and EADs in all three transmural regions. CONCLUSION: Our results support the hypothesis that a prominent I(Ks) is responsible for the ability of epicardium and endocardium to resist some but not all of the arrhythmogenic effects of I(Kr) block. The data highlight the critical importance of I(Ks) in the canine heart and the significant role of electrotonic interactions in minimizing the development of an arrhythmogenic substrate when repolarization reserve is reduced. PMID- 11900295 TI - Kearns-Sayre syndrome: association with long QT syndrome? AB - Kearns-Sayre syndrome has associated cardiac findings, predominantly complete heart block, which has been implicated as a mechanism of sudden death in these individuals. A patient with Kearns-Sayre syndrome who had syncope and multiple cardiac arrests due to ventricular tachycardia in the setting of QT prolongation is described. Long QT syndrome is implicated as another possibility for sudden death in Kearns-Sayre syndrome. This potential association is unexplained, but now totals two reports in the small numbers of patients reported. PMID- 11900296 TI - Tachycardia and bradycardia coexisting in the same pulmonary vein. AB - During segmental isolation of the pulmonary veins (PVs) in a patient with paroxysmal atrial fibrillation, there was a PV bradycardia that was dissociated from the left atrium in a segment of the right superior PV, whereas the remaining segments showed passive activation of a PV fascicle during sinus rhythm. Rapid atrial pacing induced a PV tachycardia in the nonisolated fascicles, and the dissociated PV bradycardia persisted in a segment of the same PV. These observations indicate that PV fascicles are insulated from each other and that a dissociated PV rhythm does not necessarily indicate complete isolation of a PV. PMID- 11900297 TI - Focal atrial fibrillation: A four-headed Lernaean hydra? PMID- 11900298 TI - Class I antiarrhythmic drug and coronary vasospasm-induced T wave alternans and ventricular tachyarrhythmia in a patient with Brugada syndrome and vasospastic angina. AB - A 50-year-old man presented with a history of transient chest pain and palpitations. The 12-lead ECG at rest showed normal sinus rhythm. A slight ST segment elevation was observed in leads V1 to V3. During hospitalization, atrial fibrillation developed, and oral pilsicainide was administered. Thirty minutes after the drug was given, the ECG showed marked ST segment elevation in leads V1 to V3, and T wave alternans became visible in leads V2 and V3. Self-terminating ventricular tachycardia was initiated following frequent ventricular premature complexes, which showed a left bundle branch block pattern. The coronary angiogram was normal, but in the provocation test of vasospastic angina, acetylcholine administration into the left coronary artery resulted in complete occlusion of the left anterior descending and circumflex arteries. Marked ST segment elevation developed in leads I, aVL, and V3 to V6 concomitant with visible QT/T alternans in leads V4 and V5, and ventricular tachyarrhythmia was initiated. Brugada syndrome and vasospastic angina coexisted in this patient, and T wave alternans can be used as a predictor of ventricular tachyarrhythmias in such patients. PMID- 11900299 TI - Calmodulin and the philosopher's stone: Changing Ca2+ into arrhythmias. AB - Calmodulin and the Philosopher's Stone. It has been recognized for some time that intracellular Ca2+ ([Ca2+]i) can contribute to the genesis of cardiac arrhythmias, but understanding of the molecular signaling machinery that links disordered [Ca2+]i to arrhythmias has been lacking. Exciting recent work has focused on the ubiquitous intracellular Ca2+-binding protein calmodulin. Calmodulin is a molecular sensor that can translate increases in [Ca2+]i into modulatory signals for ion channels and activate other Ca2+-dependent signaling molecules. This article will examine the implications of these recent findings for arrhythmogenesis and the development of new antiarrhythmic therapies. PMID- 11900300 TI - "Cough drops". PMID- 11900301 TI - Entrainment of ventricular tachycardia by sinus rhythm. PMID- 11900302 TI - Unusual response to atrial extrastimulus testing. PMID- 11900303 TI - A patient with polymorphic ventricular tachycardia (VT) controlled by beta blockers and atrial pacing. PMID- 11900304 TI - Is subjective shortening in human memory unique to time representations? AB - Three experiments compared forgetting of the duration of a bar-like visual stimulus with forgetting of its length. The main aim of the experiments was to investigate whether subjective shortening (a decrease in the subjective magnitude of a stimulus as its retention interval increased) was observable in length judgements as well as in time judgements, where subjective shortening has been often observed previously. On all trials of the three experiments, humans received two briefly presented coloured bars, separated by a delay ranging from 1 to 10 s, and the bars could differ in length, duration of presentation, or both. In Experiment 1 two groups of subjects made either length or duration judgements, and subjective shortening-type forgetting functions were observed only for duration. Experiments 2 and 3 used the same general procedure, but the stimuli judged could differ both in length and duration within a trial, and different subject groups (Experiment 2) or the same subjects in two conditions (Experiment 3) made either length or duration judgements of stimuli, which were on average physically identical. Subjective shortening was only found with duration, and never with length, supporting the view that subjective shortening may be unique to time judgements. PMID- 11900305 TI - Influence of dorsolateral periaqueductal grey (dlPAG) lesions on contextual conditioning with massed and distributed shock. AB - Three experiments investigated the conditions under which electrolytic lesions of the dorsolateral periaqueductal grey (dlPAG) facilitate conditioned defensive freezing in the rat (Rattus norvegicus). Experiment 1 found that dlPAG lesions placed before context-shock pairings facilitated conditioned defensive freezing with massed but not distributed shock. No such effect was found in Experiment 2, when the lesions were placed after context-shock pairings. Experiment 3 found that dlPAG lesions facilitated subsequent conditioning with massed but not a single shock. In addition, no differences in sensitivity to thermal or shock pain were evident in lesioned and unlesioned rats. Taken together, these results are consistent with the suggestion that dlPAG activation interferes with the processing of contextual cues during association formation. PMID- 11900306 TI - The location and interpretation of the bisection point. AB - In a temporal bisection task with humans, the observer is required to decide whether a probe duration (t) is more similar to the short referent (S), an R(S) response, or to the long referent (L), an RL response. Temporal bisection yields a psychometric function relating the proportion of long responses, P(R(L)), to probe duration t. The value of t at which R(S) and R(L) occur with equal frequency, P(R(L)) = .5, is referred to as the bisection point, T1/2. Bisection models usually interpret T1/2 as identifying the value of t that is equally confusable with S and L, but they differ in their predictions for the location of T1/2. The present paper presents new data relevant to the location and interpretation of T1/2. The data indicate that the empirical values usually are biased, the biases being influenced by duration range, L:S ratio, and probe spacing. Moreover, the biases often are not consistent across observers. It is concluded that empirical values of T1/2 should not be interpreted as indicating the value of t that is equally confusable with S and L. PMID- 11900307 TI - Summation: further assessment of a configural theory. AB - when A+ B+ training was conducted in the absence of CD+ trials. A further failure to observe abnormally strong responding during AB was found in Experiment 3 for which the training trials with A+ B+ CD+ were accompanied by trials in which C and D were separately paired with food. The results are explained in terms of a configural theory of conditioning, which assumes that responding during a compound is determined by generalization from its components, as well as from other compounds to which it is similar. PMID- 11900308 TI - An assessment of factors contributing to instrumental performance for sexual reward in the rat. AB - Male and female rats were gonadectomized, implanted in adulthood with capsules containing either testosterone propionate (TP) or cholesterol, and were trained to lever press for access to an oestrous female. When lever press performance was tested in extinction, only the male rats implanted with TP displayed significantly higher levels of responding than controls, demonstrating that lever pressing for oestrous females as a reward is sexually dimorphic. Ejaculation patterns from a separate assessment of these rats' copulatory ability were significantly correlated with their instrumental performance. In a second experiment, "masculinized" females exposed to TP postnatally and given TP implants responded in extinction at mean levels equivalent to those exhibited by adult males that were either intact or castrated with androgen replacement. These data suggest that organizational steroid exposure perinatally affects the actual reward value assigned to oestrous females in adulthood, in combination with consummatory sexual experience. PMID- 11900309 TI - Management of ischaemic heart disease in diabetic patients--is there a role for cardiac metabolic agents? AB - Diabetes mellitus (DM) increases the risk of cardiovascular events in patients with cardiovascular disease as well as in patients without any previous history of ischaemic heart disease (IHD). The management of IHD in diabetic patients remains a difficult challenge. However, some of these treatments are not as effective or well-tolerated in diabetic patients as in non-diabetic patients. An important effect of diabetes, that in turn influences cardiac function, is the switch from carbohydrate oxidation to free fatty acid and ketone oxidation. The correction of the alterations to cardiac metabolism associated with DM may represent a new approach to the management of IHD in these patients. Results obtained in anginal patients with the metabolic agent trimetazidine and in infarcted patients with glucose-insulin-potassium (GIK) are consistent with this hypothesis. PMID- 11900310 TI - A new therapeutic target in Alzheimer's disease treatment: attention to butyrylcholinesterase. AB - Alzheimer's disease (AD) is a progressive neurodegenerative disorder of the elderly, characterised by widespread loss of central cholinergic function. The only symptomatic treatment proven effective to date is the use of cholinesterase (ChE) inhibitors to augment surviving cholinergic activity. ChE inhibitors act on the enzymes that hydrolyse acetylcholine (ACh) following synaptic release. In the healthy brain, acetylcholinesterase (AChE) predominates (80%) and butyrylcholinesterase (BuChE) is considered to play a minor role in regulating brain ACh levels. In the AD brain, BuChE activity rises while AChE activity remains unchanged or declines. Therefore both enzymes are likely to have involvement in regulating ACh levels and represent legitimate therapeutic targets to ameliorate the cholinergic deficit. The two enzymes differ in location, substrate specificity and kinetics. Recent evidence suggests that BuChE may also have a role in the aetiology and progression of AD beyond regulation of synaptic ACh levels. Experimental evidence from the use of agents with enhanced selectivity for BuChE (cymserine, MF-8622) and ChE inhibitors such as rivastigmine, which have a dual inhibitory action on both AChE and BuChE, indicate potential therapeutic benefits of inhibiting both AChE and BuChE in AD and related dementias. The development of specific BuChE inhibitors and the continued use of ChE inhibitors with the ability to inhibit BuChE in addition to AChE should lead to improved clinical outcomes. PMID- 11900312 TI - Magnetic pulse treatment for knee osteoarthritis: a randomised, double-blind, placebo-controlled study. AB - We assessed the efficacy and tolerability of low-frequency pulsed electromagnetic fields (PEMF) therapy in patients with clinically symptomatic knee osteoarthritis (OA) in a randomised, placebo-controlled, double-blind study of six weeks' duration. Patients with radiographic evidence and symptoms of OA (incompletely relieved by conventional treatments), according to the criteria of the American College of Rheumatology, were recruited from a single tertiary referral centre. 75 patients fulfilling the above criteria were randomised to receive active PEMF treatment by unipolar magnetic devices (Medicur) manufactured by Snowden Healthcare (Nottingham, UK) or placebo. Six patients failed to attend after the screening and were excluded from analysis. The primary outcome measure was reduction in overall pain assessed on a four-point Likert scale ranging from nil to severe. Secondary outcome measures included the WOMAC Osteoarthritis Index (Likert scale) and the EuroQol (Euro-Quality of Life, EQ-5D). Baseline assessments showed that the treatment groups were equally matched. Although there were no significant differences between active and sham treatment groups in respect of any outcome measure after treatment, paired analysis of the follow-up observations on each patient showed significant improvements in the actively treated group in the WOMAC global score (p = 0.018), WOMAC pain score (p = 0.065), WOMAC disability score (p = 0.019) and EuroQol score (p = 0.001) at study end compared to baseline. In contrast, there were no improvements in any variable in the placebo-treated group. There were no clinically relevant adverse effects attributable to active treatment. These results suggest that the Medicur unipolar magnetic devices are beneficial in reducing pain and disability in patients with knee OA resistant to conventional treatment in the absence of significant side effects. Further studies using different types of magnetic devices, treatment protocols and patient populations are warranted to confirm the general efficacy of PEMF therapy in OA and other conditions. PMID- 11900311 TI - Pioglitazone: a review of Japanese clinical studies. AB - Pioglitazone, a new thiazolidinedione agent,has been shown to increase insulin sensitivity in clinical trials. Pioglitazone HCI was rapidly absorbed within one hour, achieved peak concentrations at 2-3 h, and was eliminated from serum at 24 36 h. Pioglitazone demonstrated dose-dependent pharmacokinetics. Food did not significantly affect the pharmacokinetic profile of pioglitazone. The pharmacokinetic profile of sulfonylurea agents was not significantly altered by concurrent administration with pioglitazone. Pharmacokinetic studies in healthy volunteers and in patients with type 2 diabetes indicated that pioglitazone may be administered once daily. In patients with type 2 diabetes, pioglitazone as monotherapy and in combination with sulfonylureas or an alpha-glucosidase inhibitor significantly reduced fasting blood glucose, HbA1c, triglycerides, and free fatty acids, and significantly increased HDL-cholesterol. Pioglitazone demonstrated either minor increases or decreases in cholesterol with no adverse effect on LDL-cholesterol. No patients experienced jaundice or ALT elevations > or = three times the upper limit of normal. Adverse events were mild and transient; all subjects returned to their baseline health status or laboratory tests upon withdrawal from, or completion of, the studies. Based upon these preliminary studies, full-scale clinical investigations were conducted in Japan, the United States, and Europe. As a result, in many countries pioglitazone has gained approval for use in patients with type 2 diabetes. PMID- 11900314 TI - Identification of sulfonamide-like adverse drug reactions to celecoxib in the World Health Organization database. AB - BACKGROUND: The selective COX-2 inhibitor celecoxib has a sulfonamide structure and is contraindicated for patients with known sulfa allergy. However, there is currently no standard available for the identification of sulfonamide-related adverse drug reactions (ADRs) and the occurrence of such ADRs with celecoxib has not been established. THE AIMS OF THIS STUDY WERE: (1) to identify the typical pattern of sulfonamide ADRs from literature and verify this pattern in the World Health Organization (WHO) ADR database; and (2) to examine whether these sulfonamide ADRs occur more frequently with celecoxib than with the non sulfonamide, COX-2 inhibitor rofecoxib. METHODS: A sulfonamide ADR pattern was derived from the most extensive textbook source of ADRs and applied to the WHO database for the three groups of sulfonamide drugs: short- and intermediate acting sulfonamides, and sulfasalazine. ADRs reported three or more times for each of these groups were included in a 'sulfonamide template' comprising 19 ADRs relating to the skin, the blood, the liver, and anaphylaxis. This template was then applied to celecoxib and rofecoxib. RESULTS: Overall, the relative reporting rate of a sulfonamide-type ADR with celecoxib was 80% higher than with rofecoxib, whether this was based on total number of reports (RR 1.8, 95% Cl 1.6-1.9) or restricted to reports that listed coxibs as the sole suspected drugs (RR 1.8, 95% Cl 1.6-1.9). There were numerically more ADRs for celecoxib than for rofecoxib in 15 of the 19 terms. Within the ADRs in the sulfonamide template, relative reporting rate of fatal reactions was 80% higher with celecoxib (RR 1.8, 95% Cl 0.9-4.0). Even though serious sulfonamide reactions are rare, their clinical impact on patient safety warrants close monitoring as more data becomes available. Physicians should be aware of possible sulfonamide allergy when prescribing celecoxib. PMID- 11900313 TI - Haemophilus influenzae type b vaccine: reconstitution of lyophilised PRP-T vaccine with a pertussis-containing paediatric combination vaccine, or a change in the primary series immunisation schedule, may modify the serum anti-PRP antibody responses. AB - Immunogenicity data obtained after primary series immunisations against Haemophilus influenzae type b (Hib), using a vaccine prepared by conjugating the capsular polysaccharide of Hib to tetanus toxoid (ActHIB), were compiled from 146 study groups comprising 85 clinical trials or vaccination programs conducted between 1987 and 1999. ActHIB was administered as a monovalent lyophilised vaccine, injected either in association with another paediatric vaccine (at separate administration sites) or in combination (where the different vaccines are mixed together in the same syringe before injection). Review of these data reveals two trends. First, PRP-T vaccine, given either alone or in combination with DTwcP, resulted in a stronger anti-PRP serum antibody response than when PRP T was combined with DTacP vaccine. Second, an accelerated (i.e. one-month interval) immunisation schedule tended to induce a poorer anti-PRP antibody response than did the more widely spaced, standard inoculation schedules. Although an in-depth analysis of these over 11000 study subjects on an individual basis with multivariate analysis or multifactorial statistical methods might eventually provide working hypotheses to fully understand these phenomenon, the various licensed, PRP-T-containing paediatric combination vaccines have proved to be clinically effective. PMID- 11900315 TI - Aromatase inhibitors. AB - The new non-steroidal and steroidal aromatase inhibitors are at least as effective as megestrol acetate (MA) as second-line hormonal agents in postmenopausal women with breast cancer. However, they are superior to MA in terms of tolerability and adverse effects. Letrozole and exemestane have been shown to be superior to MA in terms of efficacy. Furthermore, exemestane and anastrozole demonstrated a survival advantage over MA. These drugs are therefore considered established second-line hormonal agents. There is a growing body of evidence supporting the role of third-generation aromatase inhibitors as first line therapy for ER-and/or PgR-positive advanced breast cancer in postmenopausal women, and as a neoadjuvant therapy in postmenopausal women with hormone receptor positive tumours unsuitable for breast conserving surgery. Studies comparing these drugs head-to-head and with adjuvant tamoxifen are currently in progress. The potential role of these drugs in breast cancer prevention is worth investigating. PMID- 11900316 TI - Focus on clozapine. AB - Clozapine is a dibenzodiazepine derivative and a truly atypical anti-psychotic. Its therapeutic effects are probably mediated by dopaminergic and serotonergic activity. Although it appears to be the most effective antipsychotic drug for treatment-resistant schizophrenia, its general use is limited because of the risk of agranulocytosis. PMID- 11900317 TI - Effect of atorvastatin on serum creatinine levels. PMID- 11900318 TI - Limbal stem cells in health and disease. AB - Stem cells are present in all self-reviewing tissues and have unique properties. The ocular surface is made up of two distinct types of epithelial cells, constituting the conjunctival and the corneal epithelia. These epithelia are stratified, squamous and non-keratinized. Although anatomically continuous with each other at the corneoscleral limbus, the two cell phenotypes represent quite distinct subpopulations. The stem cells for the cornea are located at the limbus. The microenvironment of the limbus is considered to be important in maintaining stemness of the stem cells. They also act as a "barrier" to conjunctival epithelial cells and prevent them from migrating on to the corneal surface. In certain pathologic conditions, however, the limbal stem cells may be destroyed partially or completely resulting in varying degrees of stem cell deficiency with its characteristic clinical features. These include "conjunctivalization" of the cornea with vascularization, appearance of goblet cells, and an irregular and unstable epithelium. The stem cell deficiency can be managed with auto or allotransplantation of these cells. With the latter option, systemic immunosuppression is required. The stem cells can be expanded ex vivo on a processed human amniotic membrane and transplanted back to ocular surface with stem cell deficiency without the need of immunosuppression. PMID- 11900319 TI - The lipid layer: the outer surface of the ocular surface tear film. AB - The outer layer of the tear film--the lipid layer--has numerous functions. It is a composite monolayer composed of a polar phase with surfactant properties and a nonpolar phase. In order to achieve an effective lipid layer, the nonpolar phase, which retards water vapor transmission, is dependent on a properly structured polar phase. Additionally, this composite lipid layer must maintain its integrity during a blink. The phases of the lipid layer depend on both lipid type as well as fatty acid and alcohol composition for functionality. Surprisingly, the importance of the composition of the aqueous layer of the tear film in proper structuring of the lipid layer has not been recognized. Finally, lipid layer abnormalities and their relationship to ocular disease are beginning to be clarified. PMID- 11900320 TI - Keratitis. AB - Corneal inflammation or keratitis is a significant cause of ocular morbidity around the world. Fortunately, the majority of the cases are successfully managed with medical therapy, but the failure of therapy does occur, leading to devastating consequences of either losing the vision or the eye. This review attempts to provide current information on most, though not all, aspects of keratitis. Corneal inflammation may be ulcerative or nonulcerative and may arise because of infectious or noninfectious causes. The nonulcerative corneal inflammation may be confined to the epithelial layer or to the stroma of the cornea or may affect both. For clarity, this section has been divided into nonulcerative superficial keratitis and nonulcerative stromal keratitis. While the former usually includes hypersensitivity responses to microbial toxins and unknown agents, the latter can be either infectious or noninfectious. In the pathogenesis of ulcerative keratitis, microorganisms such as bacteria, fungi, parasites (Acanthamoeba), or viruses play an important role. Approximately, 12.2% of all corneal transplantations are done for active infectious keratitis. Available world literature pertaining to the incidence of microbial keratitis has been provided special place in this review. On the other hand, noninfectious ulcerative keratitis can be related to a variety of systemic or local causes, predominantly of autoimmune origin. PMID- 11900321 TI - Contact lens related corneal infections. AB - This article describes microbial keratitis, infection of the cornea by micro organisms. Contact lens wear is a predisposing factor for the development of microbial keratitis. Micro-organisms probably adhere to the contact lens, transfer from the contact lens to a damaged or compromised corneal epithelial surface, penetrate into the deeper layers of the cornea and produce corneal damage. Host responses to the invading micro-organisms, while designed to protect the eye, can often exacerbate the situation and the resulting microbial keratitis can lead to permanent blindness. The microbial, biochemical and immunological aspects of MK will be described in detail. PMID- 11900322 TI - Host-defense mechanism of the ocular surfaces. AB - The defense of the ocular surfaces presents an unique challenge in that not only must integrity be maintained against microbial, inflammatory and physical assault, but it must be done while minimizing the risk of loss of corneal transparency. This puts severe limitations on the degree to which scarring or neovascularization can occur in the cornea secondary to any infectious, inflammatory, immunological or wound healing process. Moreover, this defense system must be equally effective under two extremes of conditions: those found in the open eye and the closed eye environments. It is our contention that these constraints have resulted in the evolution of a highly complex fail-safe defense system that utilizes distinctly different strategies in open and closed eye conditions. The extraordinary effectiveness of this system is evidenced by the fact that despite continued exposure to a microbe rich environment, the external ocular surfaces maintain a very low microbial titer and are highly resistant to breaching by all but a few pathogens. It is the intent of this review to provide a working model of this defense system as it operates under both open and closed eye conditions, to provide evidence in support of this model as well as highlight some of the many areas of uncertainty. PMID- 11900323 TI - Amniotic membrane transplantation for ocular surface reconstruction. AB - The use of amniotic membrane (or amnion) for transplantation as graft in ocular surface reconstruction is reviewed. This technique has become widespread because of the availability of the amnion, convenience and ease of use, and high and reproducible success rates. The mechanisms of action of the transplantation are varied and include the prolongation and clonogenic maintenance of epithelial progenitor cells, promotion of goblet and non-goblet cell differentiation, exclusion of inflammatory cells with anti-protease activities, suppression of Transforming Growth Factor Beta signaling and myoblast differentiation of normal fibroblasts. The observed clinical effects include facilitation of epithelialization, maintenance of normal phenotypes, and reduction of inflammation, vascularization and scarring. Amniotic membrane transplantation is being increasingly used as graft for various conjunctival and corneal diseases and as a patch in cases of chemical and thermal burns, refractory and recalcitrant keratitis, and most recently as an excellent substrate for expanding epithelial stem cells ex vivo. PMID- 11900324 TI - Laser refractive surgery: technological advance and tissue response. AB - Photorefractive keratectomy (PRK) and laser assisted in situ keratomileusis (LASIK), using an excimer laser, are the currently popular techniques of correcting refractive errors. Since these techniques work by selective ablation of corneal stroma, the tissue healing response plays a great role in the ultimate outcome of surgery. Also, various methods of wound healing modulation can be used to achieve better results. While these procedures do lead to a decrease in dioptric power and increase in unaided visual acuity, higher visual functions like contrast sensitivity can sometimes be compromised after the surgery. PMID- 11900325 TI - Artificial cornea: towards a synthetic onlay for correction of refractive error. AB - Synthetic onlays that are implanted onto the surface of the cornea have the potential to become an alternative to spectacles and contact lenses for the correction of refractive error. A successful corneal onlay is dependent on development of a biocompatible polymer material that will maintain a healthy cornea after implantation and that will promote growth of corneal epithelial cells over the onlay, and development of a method for attachment of the onlay with minimal surgical invasiveness. The ideal onlay should be made of a material that is highly permeable yet has sufficient surface characteristics to stimulate stable and firm attachment of the corneal epithelium over the onlay. Recent research indicates that collagen I coated polymer materials that mimic the basement membrane of the corneal epithelium promote the most favorable growth of epithelial cells in vivo in comparison to wholly biological or synthetic materials. PMID- 11900326 TI - The ocular lens epithelium. AB - An adult lens contains two easily discernible, morphologically distinct compartments, the epithelium and the fiber-cell mass. The fiber-cell mass provides the lens with its functional phenotype, transparency. Metabolically, in comparison to the fiber cells the epithelium is the more active compartment of the ocular lens. For the purposes of this review we will only discuss the surface epithelium that covers the anterior face of the adult ocular lens. This single layer of cells, in addition to acting as a metabolic engine that sustains the physiological health of this tissue, also works as a source of stem cells, providing precursor cells, which through molecular and morphological differentiation give rise to fiber cells. Morphological simplicity, defined developmental history and easy access to the experimenter make this epithelium a choice starting material for investigations that seek to address universal questions of cell growth, development, epithelial function, cancer and aging. There are two important aspects of the lens epithelium that make it highly relevant to the modern biologist. Firstly, there are no known clinically recognizable cancers of the ocular lens. Considering that most of the known malignancies are epithelial in origin this observation is more than an academic curiosity. The lack of vasculature in the lens may explain the absence of tumors in this tissue, but this provides only a teleological basis to a very important question for which the answers must reside in the molecular make-up and physiology of the lens epithelial cells. Secondly, lens epithelium as a morphological entity in the human lens is first recognizable in the 5th-6th week of gestation. It stays in this morphological state as the anterior epithelium of the lens for the rest of the life, making it an attractive paradigm for the study of the effects of aging on epithelial function. What follows is a brief overview of the present status and lacunae in our understanding of the biology of the lens epithelium. PMID- 11900327 TI - Performance assessment of NAPL remediation in heterogeneous alluvium. AB - Over the last few years, more than 40 partitioning interwell tracer tests (PITTs) have been conducted at many different sites to measure nonaqueous phase liquid (NAPL) saturations in the subsurface. While the main goal of these PITTs was to estimate the NAPL volume in the subsurface, some were specifically conducted to assess the performance of remedial actions involving NAPL removal. In this paper, we present a quantitative approach to assess the performance of remedial actions to recover NAPL that can be used to assess any NAPL removal technology. It combines the use of PITTs (to estimate the NAPL volume in the swept pore volume between injection and extraction wells of a test area) with the use of several cores to determine the vertical NAPL distribution in the subsurface. We illustrate the effectiveness of such an approach by assessing the performance of a surfactant/foam flood conducted at Hill Air Force Base, UT, to remove a TCE rich NAPL from alluvium with permeability contrasts as high as one order of magnitude. In addition, we compare the NAPL volumes determined by the PITTs with volumes estimated through geostatistical interpolation of aquifer sediment core data collected with a vertical frequency of 5-10 cm and a lateral borehole spacing of 0.15 m. We demonstrate the use of several innovations including the explicit estimation of not only the errors associated with NAPL volumes and saturations derived from PITTs but also the heterogeneity of the aquifer sediments based upon permeability estimates. Most importantly, we demonstrate the reliability of the PMID- 11900328 TI - Using polymer mats to biodegrade atrazine in groundwater: laboratory column experiments. AB - Large-scale column experiments were undertaken to evaluate the potential of in situ polymer mats to deliver oxygen into groundwater to induce biodegradation of the pesticides atrazine, terbutryn and fenamiphos contaminating groundwater in Perth, Western Australia. The polymer mats, composed of woven silicone (dimethylsiloxane) tubes and purged with air, were installed in 2-m-long flow through soil columns. The polymer mats proved efficient in delivering dissolved oxygen to anaerobic groundwater. Dissolved oxygen concentrations increased from <0.2 mg l(-1) to approximately 4 mg l(-1). Degradation rates of atrazine in oxygenated groundwater were relatively high with a zero-order rate of 240-380 microg l(-1) or a first-order half-life of 0.35 days. Amendment with an additional carbon source showed no significant improvement in biodegradation rates, suggesting that organic carbon was not limiting biodegradation. Atrazine degradation rates estimated in the column experiments were similar to rates determined in laboratory culture experiments, using pure cultures of atrazine mineralising bacteria. No significant degradation of terbutryn or fenamiphos was observed under the experimental conditions within the time frames of the study. Results from these experiments indicate that remediation of atrazine in a contaminated aquifer may be achievable by delivery of oxygen using an in situ polymer mat system. PMID- 11900329 TI - Characterization of flow and transport processes within the unsaturated zone of Yucca Mountain, Nevada, under current and future climates. AB - This paper presents a large-scale modeling study characterizing fluid flow and tracer transport in the unsaturated zone of Yucca Mountain, Nevada, a potential repository site for storing high-level radioactive waste. The study has been conducted using a three-dimensional numerical model, which incorporates a wide variety of field data and takes into account the coupled processes of flow and transport in the highly heterogeneous, unsaturated fractured porous rock. The modeling approach is based on a dual-continuum formulation of coupled multiphase fluid and tracer transport through fractured porous rock. Various scenarios of current and future climate conditions and their effects on the unsaturated zone are evaluated to aid in the assessment of the proposed repository's system performance using different conceptual models. These models are calibrated against field-measured data. Model-predicted flow and transport processes under current and future climates are discussed. PMID- 11900330 TI - Modeling the effects of concentration history on the slow desorption of trichloroethene from a soil at 100% relative humidity. AB - To determine the effects of concentration history on slow sorption processes, desorption kinetic profiles for trichloroethene (TCE) were measured for a soil at 100% relative humidity subject to different exposure concentrations and exposure times. Exposure concentrations ranged from 1% to 80% of the saturation vapor pressure (Ps) for TCE, and exposure times (i.e., time allowed for sorption before desorption begins) ranged from 1 to 96 days. A spherical diffusion model based on a gamma distribution of sorption rates and a gamma distribution of desorption rates was developed and applied to the data. At 80% P/Ps, the entire gamma distributions of sorption and desorption rates were available for TCE. In accordance with a micropore filling mechanism, the fraction of these distributions available for TCE sorption decreased with decreasing P/Ps. Experimental results are consistent with a micropore-filling mechanism, where the amount of slow desorbing mass decreased with decreasing exposure time, and the fraction of slow desorbing sites filled decreased with decreasing exposure concentration. Simulation results suggest that diffusion limits the rates that micropores fill, and that rates of sorption and desorption for soil contaminated at smaller values of P/Ps are, on average, less than those at larger values of P/Ps (i.e., slow desorption rates are a function of exposure concentration). Simulation results also suggest that the model adequately describes the effects of exposure concentration and exposure time on the rates of sorption and desorption, but not on the capacity of the slow sites for TCE. This work is important because contaminant concentrations in the subsurface vary in space and time, and the proposed model represents a new and mechanistically based approach to capture the effects of this heterogeneity on slow desorption. PMID- 11900331 TI - Spatial and temporal distribution of solute leaching in heterogeneous soils: analysis and application to multisampler lysimeter data. AB - Accurate assessment of the fate of salts, nutrients, and pollutants in natural, heterogeneous soils requires a proper quantification of both spatial and temporal solute spreading during solute movement. The number of experiments with multisampler devices that measure solute leaching as a function of space and time is increasing. The breakthrough curve (BTC) can characterize the temporal aspect of solute leaching, and recently the spatial solute distribution curve (SSDC) was introduced to describe the spatial solute distribution. We combined and extended both concepts to develop a tool for the comprehensive analysis of the full spatio temporal behavior of solute leaching. The sampling locations are ranked in order of descending amount of total leaching (defined as the cumulative leaching from an individual compartment at the end of the experiment), thus collapsing both spatial axes of the sampling plane into one. The leaching process can then be described by a curved surface that is a function of the single spatial coordinate and time. This leaching surface is scaled to integrate to unity, and termed S can efficiently represent data from multisampler solute transport experiments or simulation results from multidimensional solute transport models. The mathematical relationships between the scaled leaching surface S, the BTC, and the SSDC are established. Any desired characteristic of the leaching process can be derived from S. The analysis was applied to a chloride leaching experiment on a lysimeter with 300 drainage compartments of 25 cm2 each. The sandy soil monolith in the lysimeter exhibited fingered flow in the water-repellent top layer. The observed S demonstrated the absence of a sharp separation between fingers and dry areas, owing to diverging flow in the wettable soil below the fingers. Times-to-peak, maximum solute fluxes, and total leaching varied more in high-leaching than in low-leaching compartments. This suggests a stochastic convective transport process in the high-flow streamtubes, while convection dispersion is predominant in the low-flow areas. S can be viewed as a bivariate probability density function. Its marginal distributions are the BTC of all sampling locations combined, and the SSDC of cumulative solute leaching at the end of the experiment. The observed S cannot be represented by assuming complete independence between its marginal distributions, indicating that S contains information about the leaching process that cannot be derived from the combination of the BTC and the SSDC. PMID- 11900332 TI - Scientific priorities and strategic planning for resuscitation research and life saving therapy following traumatic injury: report of the PULSE Trauma Work Group. Post Resuscitative and Initial Utility of Life Saving Efforts. AB - Traumatic injury and its sequelae remains a major, unrecognized, public health problem in North America. It is the principle cause of death in patients aged 1 44 and the overall leading cause of life years lost in the United States. Recognizing this the National Heart, Lung and Blood Institute (NHLBI), in conjunction with other federal agencies, organized a conference in June 2000 to discuss the basic and clinical research projects that could lead to improved outcomes following cardiopulmonary or post-injury resuscitation. The Post Resuscitative and Initial Utility of Life Saving Efforts (PULSE) Workshop resulted and eight workgroups were established to focus on various aspects including organ systems, pharmacology, epidemiology, and trauma. The Trauma Work group recommendations are presented in this manuscript. Despite the recognition of improved survival and outcome through advancements in trauma systems and trauma care, the National Institutes of Health (NIH) support ratio for trauma research is only 0.10 compared to 1.65 for cancer research and a remarkable support ratio of 3.51 for AIDS and HIV infection research. The successful federal HIV research program has significantly decreased the morbidity and mortality over the last ten years at a cost of 1.4 billion dollars per year. A coordinated trauma research program should aim to replicate the success achieved by such programs; however, a centralized federal "home" for trauma research does not exist. Consequently, the existing limited research support is derived from NIH institutes in addition to other federal and state agencies. This report serves to describe some of the obstacles and to outline various strategies and priorities for basic science, clinical and translational trauma resuscitation research. PMID- 11900333 TI - Chemokine activation within 24 hours after blunt accident trauma. AB - Chemokines mediate the migration of leukocytes to sites of inflammation. Changes in the plasma concentration of interleukin (IL)-8 and macrophage inflammatory protein (MIP)-1beta have not been investigated in the very early phase starting immediately after unintentional trauma. Enrolled in the study were 94 patients with multiple blunt injuries. Blood samples were collected at the scene of accident, then at regular intervals for 24 h. IL-8 and MIP-1beta plasma levels were determined by commercial test kits. Patients were grouped according to trauma severity, pattern of injury, as well as survivors vs. nonsurvivors. Serious casualties [Injury Severity Score (ISS) > or = 32] revealed a significant increase in IL-8 compared to only a slight elevation in individuals with an ISS < 32. Nonsurvivors showed a highly significant (P < 0.005) increase in IL-8 levels beginning immediately after admission. Trauma resulted in a modest activation of MIP-1beta production without differences regarding trauma severity, pattern of injury, or survival. A very strong trauma stimulus is necessary to activate IL-8 production. In contrast to MIP-1beta, IL-8 levels were significantly elevated in nonsurvivors compared to survivors. Therefore, IL-8 might be an early predictor of survival. PMID- 11900334 TI - The macrophage response to endotoxin requires platelet activating factor. AB - Platelet activating factor (PAF) is a key proinflammatory mediator of septic shock and is metabolized by PAF-acetylhydrolase (PAF-AH). Low circulating levels of PAF-AH have been associated with the development of autodestructive excessive inflammatory responses such as post-injury multiple organ failure, and recombinant PAF-AH is being studied for the prevention of acute respiratory distress syndrome (ARDS). However, the potential role of PAF as an autocrine mediator of macrophage activation is unclear. We wanted to examine the role of PAF in the endotoxin- (LPS) induced macrophage response using PAF-AH. Rabbit alveolar macrophages were stimulated with LPS (10 ng/mL) with or without PAF-AH (0.1-10 microg/mL). Supernatants were collected to measure the production of tumor necrosis factor (TNF), interleukin 8 (Il-8), and prostaglandin E2 (PGE2). Cell monolayers were assessed for procoagulant activity (PCA). TNF mRNA production was determined by Northern blot and RNA stability was assessed. Evaluation of intracellular signaling pathways for LPS included western blots for phosphorylated p38 and ERK kinases and gel shift for nuclear factor-kappaB. There was a dose-response inhibition of TNF, PCA, Il-8, and PGE2 production following pretreatment with PAF-AH. Time course studies revealed effective inhibition of TNF production with administration of PAF-AH up to 2 h after LPS challenge. TNF mRNA production was inhibited, while mRNA stability was not affected. There was no effect on the phosphorylation of p38 or ERK 1 kinases; however, the nuclear translocation of NF-kappaB was inhibited. Macrophage cytokine production in response to endotoxin is PAF dependent. This effect involves the inhibition of TNF gene transcription and concomitant inhibition of NF-kappaB. PMID- 11900335 TI - Lipopolysaccharide-induced enterocyte-derived nitric oxide induces intestinal monolayer permeability in an autocrine fashion. AB - Studies indicate that endotoxin (LPS) causes intestinal injury, increases inducible nitric oxide synthase (iNOS) activity, leads to increased NO production, and promotes bacterial translocation (BT). To investigate the mechanism by which LPS causes gut injury and to test the hypothesis that NO produced by enterocytes promotes gut injury in an autocrine fashion, rat intestinal epithelial cell (IEC-6) monolayers were tested. IEC-6 monolayers grown in a bicameral system were incubated with media or with LPS (25 microg/mL) and tested for permeability to phenol red, BT, and nitrate/nitrite (NO2/NO3) production. To determine the direct effect of NO on permeability, monolayers were incubated with the NO donor S-nitroso-acetylpenicillinamide (SNAP; 1 mM) and tested for permeability. Next, the protective effects of two NOS inhibitors (L NMMA and L-NIL) were tested. Finally, to determine if LPS-induced permeability occurs via a poly (ADP-ribose) synthetase- (PARS) dependent pathway, monolayers incubated with LPS alone or with the PARS inhibitor, INH2BP (100 microM) were tested. LPS significantly increased IEC-6 permeability to phenol red, as well as increased NO2/NO3 by 20-fold (P < 0.001) and increased BT 10-fold (P < 0.001). SNAP mimicked the effect of LPS and significantly increased both permeability to phenol red and BT. Inhibition of iNOS significantly decreased the LPS-induced increase in monolayer permeability and BT (P < 0.05). Monolayers incubated with INH2BP had significantly decreased permeability to phenol red and BT, suggesting that LPS-induced NO production increases monolayer permeability at least in part via a PARS-dependent mechanism. In summary, LPS-induced disruption of monolayer barrier function appears to be related, at least in part, to enterocyte produced NO. This supports the hypothesis that NO produced by LPS-stimulated enterocytes promotes injury in an autocrine fashion and highlights the fact that enterocytes can be a target as well as a producer of NO. PMID- 11900336 TI - L-NAME enhances microcirculatory congestion and cardiomyocyte apoptosis during myocardial ischemia-reperfusion in rats. AB - Besides necrosis, apoptosis is the other major mode of cardiomyocyte loss in ischemic cardiovascular disease. In the present study, we examined the hypothesis that nitric oxide (NO) protects myocardial function by improving myocardial microcirculation and attenuating cardiomyocyte apoptosis in a rat model of myocardial ischemia/reperfusion (MI/R). The left main coronary artery of anesthetized male rats was ligated for 40 min, followed by 4 h reperfusion. Four groups of animals were studied: sham operated control + saline; sham operated control + N(W)-nitro-L-arginine methyl ester (L-NAME); MI/R + saline; MI/R + L NAME (10 mg/kg, iv, 10 min prior to reperfusion). Results show that MI/R caused a decrease in mean arterial blood pressure (MABP), cardiac index (CI), and stroke volume index (SVI). Inhibition of NO synthesis by L-NAME attenuated plasma NO levels, but increased MABP and SVR in sham control rats and rats subjected to MI/R, and further depressed left ventricular function in rats subjected to MI/R as indicated by decreased CI and SVI. Furthermore, administration of L-NAME to rats subjected to MI/R enhanced cardiomyocyte apoptosis as indicated by a significant increase in DNA fragmentation compared to rats with MI/R alone. Histological study revealed that L-NAME caused arterial constriction and congestion of red blood cells in arteries and capillaries in the peri-ischemic areas of the hearts in rats subjected to MI/R and, interestingly, also in the sham control rats. Data suggest that the mechanism of increased reperfusion injury may be attributable to a "no-reflow" phenomenon induced by L-NAME, resulting in increased cardiomyocyte apoptosis secondary to ischemia and enhanced cytochrome-c release from mitochondria. In addition, cardiac injury may be increased due to the augmented oxygen consumption of cardiomyocytes caused by the increased SVR and afterload. These results suggest that endogenous NO may act to improve myocardial microvascular perfusion, reduce SVR, and limit cardiomyocyte apoptosis, thereby, attenuating myocardial dysfunction induced by MI/R. PMID- 11900337 TI - Induction of peritoneal sepsis increases the susceptibility of isolated hearts to a calcium paradox-mediated injury. AB - The present study was designed to test the hypothesis that induction of chronic peritoneal sepsis in rats would produce a more severe calcium paradox-mediated myocardial injury in isolated heart preparation than is seen in normal hearts, and that this would be inhibited by sucrose as in normal hearts. Male Sprague Dawley rats were made septic using 200 mg of cecal material (obtained from a donor rat) suspended in 5 mL of 5% dextrose in sterile water D5 W/kg. In septic animals, the cecal material was injected in the peritoneum, while sham-septic animals received only D5 W/kg (5 mL/kg). A third group consisting of normal rats (no surgery) group was also included. Hearts were harvested from all three groups and were subjected to a calcium paradox-mediated injury in an isolated heart preparation. Hearts were perfused with Krebs-Henseleit (KH) medium and were allowed to stabilize, followed by a perfusion with Ca2+-free KH for 10 min. After this 10-min Ca2+-free KH perfusion, rats were reperfused with KH medium for 60 min. Ca2+-free KH medium was used in control experiments, while sucrose experiments were conducted with the same medium except that 150 mM sucrose replaced 75 mM NaCl. A marked decrease in ATP and phosphocreatine occurred during Ca2+ reperfusion in all hearts in absence of sucrose. In the presence of the disaccharide, no change in high-energy phosphate (HEP) levels was observed in normal hearts, while lower ATP concentrations were seen in sham and septic hearts. Thus, sucrose did not inhibit cellular injury in sham and septic hearts as it did in normal hearts, and this might be due to a smaller HEP availability. Control studies with normal, sham, and septic hearts exhibited cessation of contractions in the absence of Ca2+, and appearance of large amounts of cytosolic protein in the effluent perfusate during Ca2+ reperfusion. With normal hearts, perfusion with sucrose caused a 96% inhibition of the total creatine kinase (CK) release observed in control experiments. With sham hearts, 32% of CK release was inhibited by sucrose, while 68% of the CK release was attributed to stress associated with surgery performed in the sham-septic group. In septic hearts, only 8% of the CK release was inhibited by sucrose, suggesting that more severe myocardial injury occurs when septic hearts are subjected to a calcium paradox as compared to other groups. It is evident that sucrose can inhibit a small fraction of the CK release from septic hearts during the calcium paradox as compared to the large CK loss associated with sham sepsis. We have concluded that induction of sepsis made the heart more susceptible to a calcium paradox-mediated myocardial injury. PMID- 11900338 TI - Endotoxic shock alters the pharmacokinetics of lidocaine and monoethylglycinexylidide. AB - Significant hepatic dysfunction occurs following endotoxin administration. Although the metabolism of lidocaine to one of the primary metabolites of lidocaine, monoethylglycinexylidide (MEGX), has been used as a marker of hepatic function under various conditions, it remains unknown whether these compounds can be used in vivo to evaluate hepatic function in a rat model of endotoxic shock. To study this, cytochrome P450-3A4 (CYP3A4) was determined after harvesting hepatic microsomes, hepatic blood flow was determined using radioactive microspheres, and the pharmacokinetics of lidocaine and MEGX were evaluated. Adult male Sprague-Dawley rats were divided into endotoxin (45 mg/kg, intraperitoneally; n = 28) or control (n = 32) groups. The CYP3A4 was significantly reduced after endotoxic shock. Carboxylesterase (hydrolase S) content, which was used as a control for microsomal protein, was not significantly different between groups. Total hepatic blood flow was significantly decreased (36.2 +/- 8.4 mL/min/100 g tissue vs. 120.4 +/- 10.6 mL/min/100 g tissue), which was due to the decreased portal blood flow. For the lidocaine and MEGX experiment, lidocaine (2 mg/kg) was administered followed by serial blood samples collected up to 2 h for determination of serum lidocaine and MEGX concentrations. Mean arterial pressure (MAP) was recorded throughout the experiment. The MAP was significantly lower in the endotoxin treated rats vs. control 7.5 to 8 h following endotoxin administration. Serum concentrations of lidocaine were higher in endotoxic shock versus control animals at 2 h following lidocaine administration (1.5 +/- 0.13 mg/L vs. 0.11 +/- 0.03 mg/L). Similarly, MEGX concentrations were significantly higher in endotoxic shock versus control animals (0.55 +/- 0.04 mg/L vs. 0.16 +/- 0.02, respectively) under such conditions. These data demonstrate that the elimination of lidocaine and MEGX is impaired during endotoxic shock. The elevated lidocaine and MEGX concentrations are likely to be the result of primarily reduced hepatic blood flow and secondarily due to impaired CYP450, one of which was CYP3A4. The reduced elimination of MEGX concentrations is not due to decreased hepatic metabolism of the compound via carboxylesterase. The ratio of MEGX to lidocaine concentrations, which decreased significantly following endotoxic shock, appears to be a useful measure of hepatic function during endotoxic shock where profound reductions of hepatic blood flow are observed in addition to significant reductions in CYP450. The use of only MEGX concentrations in this endotoxic shock model is not useful in evaluating liver function. PMID- 11900339 TI - Protease inhibition in the intestinal lumen: attenuation of systemic inflammation and early indicators of multiple organ failure in shock. AB - Our recent evidence suggests that pancreatic digestive enzymes in the lumen of the intestine may play a major role in the production of cardiovascular stimulatory factors during splachnic artery occlusion and reperfusion. These stimulators are detected in plasma, but their origin and mechanism of production has remained uncertain. We examine here in the rat the role of pancreatic enzymes with and without administration of a serine protease inhibitor (FOY) into the lumen of the small intestine during splanchnic artery occlusion (90 min) and reperfusion (120 min). In the presence of pancreatic enzyme inhibition in the lumen of the intestine, there is significantly enhanced survival rate, lower levels of inflammatory mediator production, the femoral artery blood pressure is maintained close to control levels, and there are significantly lower levels of cell activators in plasma. These results support the hypothesis that pancreatic enzymes may escape across the brush border barrier during intestinal ischemia and thereby initiate the production of a powerful set of cytotoxic mediators. Blockade of pancreatic enzymes in the lumen of the intestine may be a tool to interfere with inflammation and early indicators of multiorgan failure. PMID- 11900340 TI - Interleukin-10 overexpression mediates phenyl-N-tert-butyl nitrone protection from endotoxemia. AB - The free radical trapping compound phenyl N-tert-butylnitrone (PBN) provides potent protection against lethal endotoxemia in rodents, but the mechanism of this protection is not well understood. The objective of this study was to show that PBN administration in lipopolysaccharide- (LPS) induced endotoxemia promotes enhanced production of endogenous interleukin 10 (IL-10), and the expressed IL-10 is a causal factor in the protection from endotoxemia. We show the amplified expression of IL-10 in liver and plasma in PBN- (150 mg/kg) plus LPS- (4 mg/kg) treated rats using ribonuclease protection assay (RPA) and ELISA. In situ hybridization was utilized to visualize the overexpression of the IL-10 gene, and ELISA was used to determine plasma IL-10 and TNFalpha levels. Plasma IL-10 showed a 3-fold increase in PBN/LPS- treated rats compared to those treated with LPS alone, and in contrast, TNFalpha level decreased by more than 90%. However, the administration of PBN alone induced no IL-10 production. Immunoneutralization of IL-10 through anti-IL-10 antibody administration to PBN/LPS-treated rats abrogated PBN's suppression of systemic nitric oxide (NO) formation, a surrogate marker for the severity of endotoxemia, indicating that IL-10 is a causal factor for the protection. In these experiments, systemic NO level was quantified using an in vivo electron paramagnetic resonance (EPR) NO-trapping technique. Gel-shift and immunohistochemical analyses indicated that the transcription factor NF kappaB was deactivated after PBN treatment, suggesting that NF-kappaB deactivation is closely involved in IL-10 overexpression. PMID- 11900341 TI - Heat stress and/or endotoxin effects on cytokine expression by human whole blood. AB - Immune system cytokines induce vascular shock. Tumor necrosis factor-alpha (TNF alpha), interleukin 1beta (IL-1beta), and bacterial endotoxin (E) circulate in human heatstroke to suggest that E release from a heat-damaged gut may stimulate cytokines that contribute to hypovolemia. However, immune activation by heat induced tissue necrosis might stimulate cytokine generation in the absence of E. To evaluate this potential and heat stress effects on the anti-inflammatory cytokines, IL-1 receptor antagonist (IL-1ra) and IL-1 soluble receptor II (IL 1srII), a human whole blood (HWB) model was employed in which the presence or absence of E could be controlled. Using thermoelectric technology to regulate the HWB heat exposures, the temperature modulations of lethal heatstroke were precisely replicated (maximum temperature = 42.4 degrees C +/- 0.04 degrees C; thermal area = 52.3 degrees C +/- 1.5 degrees C per min). Cytokine and mRNA measurements employed enzyme-linked immunosorbant-based assay systems. Significant elevations in TNF-alpha, IL-1beta, interleukin 6 (IL-6), and IL-1ra resulted when HWB was exposed to E concentrations (10 ng/ml) reported to circulate in heatstroke. While E-stimulated IL-1ra was significantly decreased by the presence of prior heat stress (PPHS), E-stimulated IL-1beta, TNF-alpha, and IL-6 were not significantly altered by PPHS, but tended to be elevated. IL-1srII expression was unchanged by PPHS and/or E. PPHS in the absence of E did not induce cytokine responses, nor were there elevations in TNF-alpha or IL-1beta mRNA. Thus, some factor normally absent under in vitro conditions, like endotoxin, was required to provoke HWB cytokine expressions and the heat stress and E conditions that characterize heatstroke affected HWB cytokine metabolism to favor a proinflammatory environment. PMID- 11900342 TI - Beneficial effects of GPI 6150, an inhibitor of poly(ADP-ribose) polymerase in a rat model of splanchnic artery occlusion and reperfusion. AB - The aim of the present study was to investigate the effects of GPI 6150, a new poly(ADP-ribose) polymerase (PARP) inhibitor, in the pathogenesis of splanchnic artery occlusion (SAO) shock. SAO shock was induced in rats by clamping both the superior mesenteric artery and the celiac trunk for 45 min, followed by reperfusion. At 60 min after reperfusion, SAO-shocked rats developed a significant fall in mean arterial blood pressure, significant increase of tissue myeloperoxidase activity (111 +/- 4.3 U/100 mg wet tissue vs. 28 +/- 3.2 U/100 mg wet tissue of sham-operated rats), and marked histological injury to the distal ileum and a significant mortality (0% survival at 2 h after reperfusion). Immuno histochemical examination demonstrated a marked increase in the immunoreactivity to PARP, P-selectin, and intercellular adhesion molecule (ICAM-1) in the necrotic ileum. GPI 6150 treatment significantly improved mean arterial blood pressure, prevented the infiltration of neutrophils (72 +/- 3.6 U/100 mg wet tissue) into the reperfused intestine, improved the histological status of the reperfused tissues, markedly reduced the intensity of P-selectin and ICAM-1 in tissue section from SAO-shocked rats, and improved survival. In conclusion, our study demonstrates that GPI 6150 exerts multiple protective effects in splanchnic artery occlusion/reperfusion shock. PMID- 11900343 TI - Enalaprilat improves systemic cardiovascular parameters and mesenteric blood flow during hypotensive resuscitation from hemorrhagic shock in dogs. AB - Resuscitative interventions that improve mesenteric perfusion without causing instability in systemic arterial pressures may be helpful for improving trauma patient outcomes. Blocking angiotensin II formation with enalaprilat may be such an intervention. Two questions were addressed in this two-part study investigating resuscitation from hemorrhagic shock in dogs: Can systemic arterial pressures be maintained while administering a constant rate infusion of enalaprilat during resuscitation, and can enalaprilat improve cardiovascular status during resuscitation? Animals were hemorrhaged to a mean arterial pressure (MAP) of 40 to 45 mmHg for 30 min and then 30 to 35 mmHg for 30 min. Group I (n = 5) was resuscitated to a MAP 60 to 65 mmHg with enalaprilat (0.02 mg/kg/h). Group II was resuscitated to a MAP 40 to 45 mmHg with (n = 5) or without (n = 5) enalaprilat. Resuscitation in both groups consisted of intermittent intravenous lactated Ringer's solution (60 mL/kg/h) to reach and maintain the target MAPs. Systemic arterial pressures were unaffected by enalaprilat during resuscitation in Group I, allowing us to proceed to the second study. During severely hypotensive resuscitation (Group II), systemic arterial pressures were also stable and enalaprilat administration was associated with increases (P < or = 0.02) in cardiac index (+1.2 L/min/m2), stroke volume index (SVI) (+14.5 mL/m2), superior mesenteric artery flow (+80 mL/min), stroke work (+561 mmHg/mL/m2), and left ventricular power output (+55.7 mmHg/L/min/m2). Corresponding increases were not observed in controls. We conclude that administration of a constant rate infusion of enalaprilat during resuscitation can be accomplished without causing a hypotensive crisis. Since enalaprilat significantly improved cardiovascular status including mesenteric perfusion even during intentional hypotension, it has potential value for improving the treatment of trauma patients. PMID- 11900344 TI - Resuscitation from severe hemorrhagic shock after traumatic brain injury using saline, shed blood, or a blood substitute. AB - The original purpose of this study was to compare initial resuscitation of hemorrhagic hypotension after traumatic brain injury (TBI) with saline and shed blood. Based on those results, the protocol was modified and saline was compared to a blood substitute, diaspirin cross-linked hemoglobin (DCLHb). Two series of experiments were performed in anesthetized and mechanically ventilated (FiO2 = 0.4) pigs (35-45 kg). In Series 1, fluid percussion TBI (6-8 ATM) was followed by a 30% hemorrhage. At 120 min post-TBI, initial resuscitation consisted of either shed blood (n = 7) or a bolus of 3x shed blood volume as saline (n = 13). Saline supplements were then administered to all pigs to maintain a systolic arterial blood pressure (SAP) of >100 mmHg and a heart rate (HR) of <110 beats/min. In Series 2, TBI (4-5 ATM) was followed by a 35% hemorrhage. At 60 min post-TBI, initial resuscitation consisted of either 500 mL of DCLHb (n = 6) or 500 mL of saline (n = 5). This was followed by saline supplements to all pigs to maintain a SAP of >100 mmHg and a HR of <110 beats/min. In Series 1, most systemic markers of resuscitation (e.g., SAP, HR, cardiac output, filling pressures, lactate, etc.) were normalized, but there were 0/7 vs. 5/13 deaths within 5 h (P = 0.058) with blood vs. saline. At constant arterial O2 saturation (SaO2), mixed venous O2 saturation (SvO2), cerebral perfusion pressure (CPP), and cerebral venous O2 saturation (ScvO2) were all higher, intracranial pressure (ICP) was lower, and CO2 reactivity was preserved with blood vs. saline (all P < 0.05). In Series 2, SAP, ICP, CPP, and lactate were higher with DCLHb vs. saline (all P< 0.05). Cardiac output was lower even though filling pressure was markedly elevated with DCLHb vs. saline (both P< 0.05). Neither SvO2 nor cerebrovascular CO2 reactivity were improved, and ScvO2 was lower with DCLHb vs. saline (P < 0.05). All survived at least 72 h with neuropathologic changes that included sub-arachnoid hemorrhage, midline cerebellar necrosis, and diffuse axonal injury. These changes were similar with DCLHb vs. saline. Thus, whole blood was more effective than saline for resuscitation of TBI, whereas DCLHb was no more, and according to many variables, less effective than saline resuscitation. These experimental results are comparable to those in a recent multicenter trial using DCLHb for the treatment of severe traumatic shock. Further investigations in similar experimental models might provide some plausible explanations why DCLHb unexpectedly increased mortality in patients. PMID- 11900345 TI - Mechanisms of action of ribavirin in antiviral therapies. AB - Although ribavirin was originally synthesized over 30 years ago and has been used to treat viral infections as monotherapy (respiratory syncytial virus and Lassa fever virus) or with interferon-alpha (IFN-alpha) as combination therapy (hepatitis C virus), the precise mechanism of its therapeutic activities remains controversial. In this review we focus on two main biological properties of ribavirin: its indirect and direct antiviral activities (with particular emphasis on its efficacy against chronic hepatitis C infection). Each property could individually or collectively account for its clinical efficacy against viral infections. First, with emphasis on the evidence for indirect activities of ribavirin, we will review the clinical observations that suggest that the immunomodulatory properties of ribavirin can in part account for its antiviral activities in vivo. We will then describe the mode of ribavirin's direct antiviral activities. These direct activities can be ascribed to several possible mechanisms, including the recently described activity as an RNA mutagen, a property that may be important in driving a rapidly mutating RNA virus over the threshold to 'error catastrophe'. PMID- 11900346 TI - Peptidyl diazomethyl ketones inhibit the human rhinovirus 3C protease: effect on virus yield by partial block of P3 polyprotein processing. AB - The efficacy of a series of diazomethyl ketones (DMKs) was measured in rhinovirus infected cultures and against the HRV14 3C protease. Their specificity and potency were confirmed against purified recombinant enzyme expressed in a yeast secretion system. An internally quenched fluorescent peptide substrate was used to assess the potency against the enzyme, obtaining a 50% inhibitory concentration (IC50) of 1 microM for both Z-L-F-Q-CHN2 and Z-V-L-F-Q-CHN2, while a lower affinity was observed for Z-F-Q-CHN2. The tripeptide Z-L-F-Q-CHN2 blocked viral replication with an IC50 value of 30 microM as judged by the reduction in viral induced cytopathy of HeLa-H1 cells, as well as a marked reduction in viral plaque formation (50% effective concentration=20 microM). Western blot analysis of viral proteins from infected cells indicates that this inhibitor works specifically by blocking viral polyprotein maturation, displaying a reduction of detectable 3C protease and an accumulation of the 3CD polypeptide. These results indicate that DMK inhibitors of the 3C protease have antiviral potency. Furthermore, the pattern of viral protein processing observed suggests that reducing the concentration of mature HRV 3C protease even in the presence of increased 3CD protein is sufficient to block proper viral processing and significantly reduce virus yield. PMID- 11900347 TI - Antipoliovirus flavonoids from Psiadia dentata. AB - The search for antiviral agents against vesicular stomatitis virus, herpes simplex virus type 1 and poliovirus type 2 in plants extracts, led to the isolation of two antipoliovirus flavonoids from the medicinal plant Psiadia dentata (Cass.) DC, Asteraceae: 3-methylkaempferol and 3,4'-dimethylkaempferol. The antipoliovirus activity of both compounds was estimated by comparison with 3 methylquercetin, guanidine and Ro-090179. The most potent inhibitor of poliovirus replication was 3-methylkaempferol, and therefore we investigated its mechanism of action. We showed, using the inhibition of [3H]uridine incorporation in viral RNA and performing a dot-blot with one RNA probe specific for the poliovirus genomic strand RNA, that 3-methylkaempferol inhibits the genomic RNA synthesis of poliovirus. PMID- 11900348 TI - Synthesis and antiviral evaluation of phosphoramidate derivatives of (E)-5-(2 bromovinyl)-2'-deoxyuridine. AB - We report the design, synthesis and antiviral evaluation of a number of lipophilic, masked phosphoramidate derivatives of the antiherpetic agent (E)-5-(2 bromovinyl)-2'-deoxyuridine (BVDU), designed to act as membrane soluble prodrugs of the free nucleotide. The phosphoramidate derivatives of BVDU that contain L alanine exhibited potent anti herpes simplex virus type 1 and varicella-zoster virus activity but lost marked activity against thymidine kinase-deficient virus strains. The phosphoramidate derivative bearing the amino acid alpha,alpha dimethylglycine showed poor activity in all cell lines tested. It appears that successful kinase bypass by phosphoramidates is highly dependent on the nucleoside analogue, amino acid and ester structure, as well as the cell line to which the drugs are exposed. PMID- 11900349 TI - Antiviral activity of cyclosaligenyl prodrugs of acyclovir, carbovir and abacavir. AB - The cyclosaligenyl (cycloSal) derivatives of the monophosphates of three acyclic or carbocyclic guanosine analogues, for example, acyclovir (ACV), carbovir (CBV) and abacavir (ABC), were investigated for their activity against retrovirus (HIV, Moloney sarcoma virus) and herpes simplex virus (HSV) activity in cell culture. The extent of the antiviral potency of the prodrugs depended on the nature of the nucleoside, the substituent on the cycloSal moiety and the virus investigated. Most notably, and unlike the parent compound ACV, cycloSal-ACV monophosphate (MP) prodrugs retained pronounced activity against ACV-resistant (thymidine kinase deficient) HSV-1 and also gained anti-HIV activity. While the cycloSal-CBVMP prodrugs did not show enhanced activity compared with the parent compound CBV, the cycloSal-ABCMP prodrugs afforded markedly increased potency against both HSV and HIV. Our data indicate that the cycloSal prodrug approach can be useful to deliver directly the MP derivatives of nucleoside analogues into the intact, virus-infected cells, thus improving and extending the antiviral potency and spectrum of the drugs against retro- and herpesvirus strains. PMID- 11900350 TI - Anti-HIV-1 activity and structure-activity relationship of cepharanoline derivatives in chronically infected cells. AB - Cepharanthine (12-O-methyl cepharanoline) is a plant alkaloid and has been shown to inhibit tumour necrosis factor-alpha- or phorbol 12-myristate 13-acetate induced HIV-1 replication in the chronically infected promonocytic cell line, U1. Its mechanism of action is considered to be the inhibition of nuclear factor kappaB, a potent inducer of HIV-1 gene expression. In this study, we have synthesized 96 derivatives of cepharanoline, including cepharanthine, and examined their inhibitory effects on HIV-1 replication in U1 cells. Among the 12 O-alkyl derivatives, cepharanthine proved to be the most active, and the activity decreased as the length of the alkyl chain increased. All of the 12-O-acyl derivatives were totally inactive, while a few 12-O-carbamoyl derivatives displayed modest activity. Since 12-O-ethyl derivatives were found to be as active as cepharanthine against HIV-1 replication, we further synthesized various 12-O-ethyl derivatives of cepharanoline. Among the derivatives, five proved to be more active inhibitors than cepharanthine, and the most active compound was 12-O ethylpiperazinyl cepharanoline. The 50% effective concentrations of this compound and cepharanthine were 0.0041 and 0.028 microg/ml (0.0060 and 0.046 microM), respectively. PMID- 11900352 TI - Point-of-purchase tobacco environments and variation by store type--United States, 1999. AB - To promote its products, the tobacco industry spent $8.2 billion on marketing in 1999, an increase of $1.5 billion over the previous year. Tobacco advertising in various media increases tobacco consumption and adolescents are more susceptible than adults to being influenced by some forms of tobacco advertising. To describe the retail tobacco advertising and marketing environment, researchers from the Robert Wood Johnson Foundation-sponsored ImpacTeen Project collected and analyzed store observation data in 163 communities throughout the United States. This report summarizes the extent of point-of-purchase (POP) tobacco advertising and marketing found in various types of stores. The findings in this report indicate that certain retail environments frequented by teenagers heavily promote tobacco use. To reduce demand for tobacco products among adolescents, public health efforts should address POP environment exposure to tobacco advertising and marketing. PMID- 11900351 TI - Tularemia--United States, 1990-2000. AB - Tularemia is a zoonotic disease caused by the gram-negative coccobacillus Francisella tularensis. Known also as "rabbit fever" and "deer fly fever," tularemia was first described in the United States in 1911 and has been reported from all states except Hawaii. Tularemia was removed from the list of nationally notifiable diseases in 1994, but increased concern about potential use of F. tularensis as a biological weapon led to its reinstatement in 2000. This report summarizes tularemia cases reported to CDC during 1990-2000, which indicate a low level of natural transmission. Understanding the epidemiology of tularemia in the United States enables clinicians and public health practitioners to recognize unusual patterns of disease occurrence that might signal an outbreak or a bioterrorism event. PMID- 11900353 TI - Variation in homicide risk during infancy--United States, 1989-1998. AB - Homicide is the 15th leading cause of death during the first year of life (i.e., infancy) in the United States. In addition, the risk for homicide is greater in infancy than in any other year of childhood before age 17 years and is greatest during the first 4 months of life. To determine how the risk for homicide varied by week during infancy and by day during the first week of life, CDC analyzed death certificate data for 1989-1998. This report summarizes the results of this analysis, which indicated that risk for infant homicide is greatest on the day of birth. Efforts to prevent infant homicides should focus on early infancy. PMID- 11900354 TI - Manufacturer's recall of rapid assay kits based on false positive Cryptosporidium antigen tests--Wisconsin, 2001-2002. AB - The Wisconsin Division of Public Health and the Wisconsin State Laboratory of Hygiene (WSLH) reported that a recent cluster of cryptosporidiosis cases in a three-county area in southeastern Wisconsin was the result of false-positive tests. During December 1, 2001-February 1, 2002, approximately 30 cases of cryptosporidiosis were diagnosed at a laboratory in southeastern Wisconsin using the Becton, Dickinson, and Company (Franklin Lakes, New Jersey) ColorPAC Cryptosporidium/Giardia rapid assay (lot number 219370, expiration date 2002-06 05). Seventeen stool specimens, which were collected from 11 patients and tested positive by the rapid assay, were re-evaluated at WSLH. Six of these stool specimens were in EcoFix (Meridian Bioscience Inc., Cincinnati, Ohio), eight were in Cary-Blair transport media, and three were formalin fixed. All 17 specimens tested negative for Cryptosporidium at WSLH using the hot safranin stain and MeriFluor (Meridian Bioscience Inc., Cincinnati, Ohio) Cryptosporidium/Giardia direct fluorescent antibody kit with concentrated specimens. PMID- 11900355 TI - Type 2 diabetes in youth: a new epidemic. PMID- 11900356 TI - Diabetes and endothelial function: implications for coronary angioplasty. PMID- 11900357 TI - Genes, environment and diabetes in Canadian aboriginal communities. PMID- 11900358 TI - The effect of hypertension in diabetes. PMID- 11900359 TI - The diabetic hypertensive (or hypertensive diabetic)--a compelling need to optimize blood pressure. AB - In summary, the present information on treating hypertension in the diabetic overwhelmingly indicates a compelling need to lower BP to target diastolic BP of 80 mmHg or less, to be less concerned about the types of drugs used than the blood pressures achieved and the concordance with therapy and to rely on two or more antihypertensive drugs in the majority of cases. Management of the hypertensive diabetic is very cost-effective. It is clear that we must engage in total cardiovascular risk management if we are to prevent the microvascular and macrovascular complications in the hypertensive diabetic (or the diabetic hypertensive). PMID- 11900360 TI - Left ventricular diastolic dysfunction in diabetic or hypertensive subjects: role of collagen alterations. PMID- 11900361 TI - Alterations in the vascular actions of insulin in the pathogenesis of insulin resistance and hypertension. PMID- 11900362 TI - Cardiac hypertrophy in diabetes patients with and without hypertension: effects of troglitazone, a novel antidiabetic drug, on diastolic function. PMID- 11900363 TI - Cost and benefits of blood pressure monitoring and control. PMID- 11900364 TI - Malonyl CoA control of fatty acid oxidation in the diabetic rat heart. AB - Increased fatty acid metabolism can decrease cardiac function and efficiency, and may therefore contribute to the outcome of ischemic injury in the diabetic. Alterations in the control of myocardial malonyl CoA levels is an important contributing factor to these high fatty acid oxidation rates. This includes alterations in AMPK, ACC, and MCD activity in the diabetic rat heart. A further understanding of how malonyl CoA controls fatty acid oxidation in the diabetic heart should help identify new targets for pharmacological intervention which decreases the reliance of the heart on fatty acid oxidation, and ultimately improves heart function. PMID- 11900365 TI - Alterations in 1,2-diacylglycerols and ceramides in diabetic rat heart. PMID- 11900366 TI - Regulation of myocardial phospholipid N-methylation by insulin and diabetes. PMID- 11900367 TI - Reduction of phosphatidylinositol-4,5-bisphosphate mass in heart sarcolemma during diabetic cardiomyopathy. PMID- 11900369 TI - Antioxidants, free radical stress and diabetes. PMID- 11900370 TI - Diabetes in sub-Saharan Africa. AB - NCDs including diabetes, heart disease and stroke are global epidemics of the 21st century. The greatest burden on health will be in developing countries and sub-Saharan Africa is an area of major challenge: We are concerned with planning for the adult victims of the new epidemic and this includes the development of appropriate treatment. Therapy should be cost effective and evidence on the economics of treating chronic conditions in Africa is urgently required. Finally, health promotion, primary prevention and health screening strategies for chronic diseases such as diabetes, stroke and coronary heart disease are required. PMID- 11900368 TI - Tumor necrosis factor-alpha, sphingomyelinase and ceramides activate tyrosine kinase, p21Ras and phosphatidylinositol 3-kinase: implications for glucose transport and insulin resistance. PMID- 11900371 TI - Oxidative stress and functional deficit in diabetic cardiomyopathy. AB - When the equilibrium between free-radical production and cellular antioxidant defences is disturbed in favour of more free radicals, it causes oxidative stress which can promote cellular injury. Oxidative stress has been suggested to play a role in the pathogenesis of diabetic cardiomyopathy. In streptozotocin-induced diabetes, there is a decrease in antioxidant enzyme activities and an increase in myocardial lipid peroxidation. Probucol, an antioxidant, was found to improve cardiac function which may have been due to an increase in myocardial antioxidant enzyme activities and a decrease in lipid peroxidation in the diabetic animals. Some of the beneficial effects of probucol may also be due to an improvement in plasma insulin levels and a decrease in the plasma glucose. The diabetic state is also associated with endothelial dysfunction, retinopathy, neuropathy and renopathy. Some of these secondary complications may also be mediated by oxidative stress. It is suggested that diabetic cardiomyopathy is associated with an antioxidant deficit and that antioxidant therapy may be useful in improving cardiac function in diabetes. PMID- 11900372 TI - Ketosis and the generation of oxygen radicals in diabetes mellitus. PMID- 11900373 TI - Effect of diabetes mellitus on hemodynamic and cardiometabolic correlates in experimental myocardial infarction. PMID- 11900374 TI - Cardiac function in perfused hearts from diabetic mice. PMID- 11900375 TI - Mg2+-dependent ATPase activity in cardiac myofibrils from the insulin-resistant JCR:LA-cp rat. AB - There is a great deal of information presently available documenting a cardiomyopathic condition in insulin-deficient models of diabetes. Less information is available documenting a similar status in non insulin-dependent models of diabetes. We have studied the functional integrity of the myofibrils isolated from hearts of JCR:LA rats. The JCR:LA rat is hyperinsulinemic, hyperlipidemic, glucose intolerant and obese. As such, it carries many of the characteristics found in humans with non insulin-dependent diabetes mellitus. These animals also have many indications of heart disease. However, it is not clear if the hearts suffer from vascular complications or are cardiomyopathic in nature. We examined Mg2+-dependent myofibrillar ATPase in hearts of JCR:LA-cp/cp rats and their corresponding control animals (+/?) and found no significant differences (P> 0.05). This is in striking contrast to the depression in this activity exhibited by cardiac myofibrils isolated from insulin-deficient models of diabetes. Our data demonstrate that myofibrillar functional integrity is normal in JCR:LA-cp rats and suggest that these hearts are not in a cardiomyopathic state. Insulin status may be critical in generating a cardiomyopathic condition in diabetes. PMID- 11900376 TI - Cyclic nucleotide phosphodiesterase families in intracellular signaling and diabetes. PMID- 11900378 TI - Coronary heart disease and risk factors in Asian Indians. PMID- 11900377 TI - Alterations in g-protein-linked signal transduction in vascular smooth muscle in diabetes. AB - The present studies were undertaken to determine the levels of stimulatory and inhibitory guanine nucleotide regulatory proteins (Gs and Gi respectively) and their relationship with adenylyl cyclase activity in aorta from 5-day streptozotocin-induced diabetic (STZ) rats. The levels of Gi alpha-2 as determined by immunoblotting techniques using AS/7 antibody were significantly decreased by about 60% in STZ as compared to control rats, whereas the levels of Gs alpha were not altered. In addition, the stimulatory effect of cholera toxin (CT) on GTP-sensitive adenylyl cyclase was not different in STZ as compared to control rats. On the other hand, the stimulatory effects of GTPgammaS, isoproterenol, glucagon, forskolin (FSK) and sodium fluoride on adenylyl cyclase were enhanced in STZ-rats. Furthermore, GTPgammaS inhibited FSK-stimulated adenylyl cyclase activity in a concentration-dependent manner (receptor independent functions of Gi) in control rats which was almost completely abolished in STZ rats. In addition, receptor-mediated inhibition of adenylyl cyclase by angiotensin II (AII), oxotremorine and atrial natriuretic peptide (ANP) was attenuated in STZ rats. These results suggest that the decreased expression of Gi alpha, but not of Gs alpha, may be responsible for the observed altered responsiveness of adenylyl cyclase to hormonal stimulation and inhibition in STZ-rats. It may thus be suggested that the decreased Gi activity may be one of the possible mechanisms responsible for the impaired vascular functions in diabetes. PMID- 11900379 TI - Mechanism of HDL lowering in insulin resistant states. PMID- 11900380 TI - Insulin treatment post myocardial infarction: the DIGAMI study. PMID- 11900381 TI - Response to ischemia and endogenous myocardial protection in the diabetic heart. PMID- 11900383 TI - The diabetic heart and cardiac glycosides. PMID- 11900382 TI - Type 2 diabetes: pharmacological intervention in an animal model. PMID- 11900384 TI - Hyperglycemia-induced protein kinase signaling pathways in vascular smooth muscle cells: implications in the pathogenesis of vascular dysfunction in diabetes. PMID- 11900385 TI - Regulation of myocardial and skeletal muscle Na,K-ATPase in diabetes mellitus in humans and animals. PMID- 11900386 TI - Food strategies for diabetes and heart health. PMID- 11900387 TI - The Atherosclerosis Reversal Clinic: the way of the future. PMID- 11900388 TI - Lipid clinics/cardiovascular risk reduction clinics: current state and future consideration. PMID- 11900389 TI - Amputation prevention and rehabilitation in diabetes. PMID- 11900390 TI - The plasma glucose level--a continuous risk factor for vascular disease in both diabetic and non-diabetic people. PMID- 11900391 TI - Laboratory testing in diabetes mellitus. PMID- 11900392 TI - Diabetes and cardiovascular disease. PMID- 11900393 TI - Prevalence of cardiovascular risk factors in Canadians with diabetes mellitus. PMID- 11900394 TI - G7 Project Promoting Heart Health. PMID- 11900395 TI - Canadian Heart Health Initiative: research review. PMID- 11900396 TI - Putting policy into action. PMID- 11900397 TI - National and international strategies to prevent obesity and diabetes. PMID- 11900398 TI - The Diabetes Council of Canada (DCC): coordinating national diabetes strategies. PMID- 11900399 TI - The federal government and diabetes: an incremental process. PMID- 11900400 TI - Modified lipoproteins and cardiovascular risk. PMID- 11900402 TI - Homocysteine as a risk factor in cardiovascular disease. PMID- 11900401 TI - Advances in lipid-lowering therapy in atherosclerosis. AB - The accrued evidence that lipid-lowering therapy limits the progression of atherosclerosis and reduces CAD events is overwhelming. The focus has been on LDL C reduction with statins, but recent evidence also stresses the importance of raising HDL-C and reducing triglyceride-rich lipoproteins (TRL). Treatment should take into account the type of dyslipidemia, combination therapy, drug interactions and pleiotropic effects of drugs (multiple effects in different systems). Statins and fibrates are the most widely prescribed. Fibrates have a major impact on plasma TRL and HDL-C levels. They enhance lipoprotein lipase, apoAI and apoAII transcription and reduce that of apoCIII. The discovery that their multiple actions are in large part mediated by the PPAR alpha pathway is a breakthrough. Fibrates also lower plasma fibrinogen and plasma viscosity but their ability to inhibit smooth muscle cell activation is one of their most promising pleiotropic effects. Statins are safe and potent LDL-C-lowering agents but also lower TRL and raise HDL. Their pleiotropic effects are numerous, and include vasodilatory, anti-thrombotic, antioxidant, anti-proliferative, anti inflammatory and plaque stabilizing properties. Many findings make a case for their early use in CAD to improve myocardial perfusion after a myocardial infarction, and they are indicated in heart transplant recipients to improve survival and reduce graft rejection. Fibrates and statins have complementary lipid modifying and pleiotropic effects so that their combination, carried out with caution to avoid potential untoward effects, should provide the highest cardiovascular benefit. This hypothesis is currently being tested in the Lipid in Diabetes Study (LDS), an outcome trial comparing monotherapy with fenofibrate and cerivastatin to combination therapy conducted in England. PMID- 11900403 TI - Endothelial integrity and repair. PMID- 11900404 TI - Undiagnosed diabetes: burden and significance in the Canadian population. PMID- 11900405 TI - Molecular mechanisms of endothelial dysfunction in the diabetic heart. AB - Our observations show that long term hyperglycaemia by the formation of AGE, but also short term hyperglycaemic periods ("glucose spikes") damage the endothelium of the heart in diabetes. The endothelium is exposed to oxidative stress. The simultaneous generation of NO and superoxide anions enables the reaction of both species to form peroxynitrite which has been identified as an important mediator for the transformation of endothelium from an anticoagulant to a procoagulant state. Together with a functional loss of endothelium these processes are assumed to impair the coronary perfusion and to provoke adaptive processes which finally lead to cardiac dysfunction and remodelling of cardiac structure (Figure 6) as it has been described for the heart in diabetes. PMID- 11900406 TI - A study of vascular wound healing in a rabbit model of type I diabetes. PMID- 11900407 TI - Endothelins in the microvasculature and heart in diabetes. PMID- 11900408 TI - Per diem rates and true costs: apples and oranges. PMID- 11900409 TI - Contributions of mouse biology to breast cancer research. PMID- 11900411 TI - Usefulness of human coagulation and fibrinolysis assays in domestic pigs. AB - Pigs are often used as animal models in research on blood coagulation and fibrinolysis. The usefulness of the assays applied within this field, and the knowledge of reference intervals are therefore essential and of utmost importance. In the study reported here, we investigated the applicability of commercial human coagulation and fibrinolysis assays for use with porcine plasma. In total, 22 functional and immunologic assays were applied to plasma obtained from domestic pigs, and the following blood coagulation and fibrinolysis variables were measured: prothrombin time, activated partial thromboplastin time, tissue factor, tissue factor pathway inhibitor, factor VII, protein C, protein S, prothrombin fragment 1+2, antithrombin, thrombin-antithrombin complexes, fibrinogen, soluble fibrin, urokinase-type plasminogen activator, plasmin inhibitor, plasminogen activator inhibitor 1, and D-dimer. We found that 11 of 12 functional assays, but only 3 of 10 immunoassays, were applicable to porcine plasma, and we determined the normal range of these variables. We conclude that human functional assays are useful in porcine plasma, whereas only a few immunologic assays can be used. However, precautions must be taken in interpretation of the results and in extrapolation toward human results because possible differences between porcine and human values can be due to species variations and/or methodologic errors. PMID- 11900410 TI - Food and secretagogue stimulation decrease the digestive enzyme content remaining in the rat pancreas. AB - The aim of the study reported here was to investigate changes in the digestive enzyme content in the pancreas after food and secretagogue stimulation. Rats from which food had been withheld overnight were either fed (between 6 and 8 a.m.) or not before euthanasia and pancreatic excision (at 8 a.m.: 21 not fed and 21 fed) and at 4 (12 p.m.: six not fed and six fed) and 8 h later (4 p.m.: six not fed and six fed). Another 16 rats were anesthetized, fitted with jugular vein and pancreatic duct catheters, and infused with the secretagogues, CCK-33 and secretin, during 1.5 h of pancreatic juice collection before euthanasia and pancreatic excision. The pancreata were homogenized, and total soluble protein and individual enzyme (trypsin and amylase) tissue contents were analyzed. Results indicated lower amounts of protein and enzymes remaining in the pancreata of the fed, compared with non-fed rats. Enzyme values indicated recovery within four hours in fed rats, but non-fed rats also had increased values during daytime. High enzyme secretion during the high dose of hormonal stimulation was reflected in lower enzyme values remaining in the pancreas, compared with that in response to low-dose stimulation. Results indicated that stimulation of the pancreas, either by food ingestion or exogenous secretagogues, lowers the amounts of digestive enzymes remaining in the pancreas, and imply that stimulation and circadian rhythms influence the pancreatic enzyme content at euthanasia. This finding should be borne in mind in interpretation of data from pancreatic studies. PMID- 11900412 TI - Comparison between two types of behavioral variables of non-evoked facial pain after chronic constriction injury to the rat infraorbital nerve. AB - BACKGROUND AND PURPOSE: Chronic constriction injury to the rat infraorbital nerve (IoN-CCI) was reported to induce asymmetric face grooming directed to the territory of the injured nerve, and localized mechanical allodynia. The model has been used for pharmacologic testing; responsiveness to mechanical stimulation has been used as outcome measure, but face grooming behavior was not studied in this context. METHODS: Face grooming data from a series of four experiments using the IoN-CCI model were retrospectively analyzed, and two types of face grooming were identified: on the one hand, isolated face grooming (i.e., face grooming that is neither preceded nor followed by body grooming); and on the other hand, face grooming during body grooming (i.e., face grooming that is part of more general body grooming behavior). RESULTS: In all four experiments, amount of isolated face grooming was found to be significantly increased after IoN-CCI. In contrast, the amount of face grooming during body grooming was not significantly altered after IoN-CCI in any of the four experiments. CONCLUSIONS: The amount of isolated face grooming is a more sensitive outcome measure of neuropathic pain than is the total amount of face grooming, which includes face grooming during body grooming. PMID- 11900413 TI - Comparative study of histopathologic characterization of azoxymethane-induced colon tumors in three inbred rat strains. AB - To obtain controlled genetic variation, colon cancer was chemically induced by use of four subcutaneous injections of azoxymethane (15 mg/kg of body weight/wk) to rats of 3 inbred strains (BDIX/OrlIco, F344/NHsd, WAG/Rij). The selection was based on the availability of established colon cancer cell lines arising from these particular strains. In the first experiment, only female rats were used; in the second experiment, both sexes were studied. The goal was to select a rat strain giving the highest tumor frequency with the shortest latency period in reproducible manner. The histologic characteristics should resemble the corresponding human tumors. The size of the tumors should be at about 1 cm in diameter, as these tumor cells were intended to be used in future transplantation studies. The two experiments yielded highly reproducible results: histologic evaluation of all colon tumors in all three rat strains revealed adenomas and adenocarcinomas closely resembling their human counterpart. The BDIX strain had the highest tumor frequency (75%) in both sexes and the shortest minimal latency period (28 weeks in experiment 1; 23 weeks in experiment 2). Tumor size of about 1 cm in diameter was found most often in the BDIX strain. On the basis of results of these two experiments, the BDIX strain has been selected for future study. PMID- 11900414 TI - A transgenic mouse strain with antigen-specific T cells (RAG1KO/sf/OVA) demonstrates that the scurfy (sf) mutation causes a defect in T-cell tolerization. AB - The scurfy (sf) murine mutation causes severe lymphoproliferation, which results in death of hemizygous males (sf/Y) by 22 to 26 days of age. The CD4+ T cells are crucial mediators of this disease. Recent publications have not only identified this mutation as the genetic equivalent of the human disease X-linked neonatal diabetes mellitus, enteropathy, and endocrinopathy syndrome, but also have indicated that the defective protein-scurfin-is a new forkhead/winged-helix protein with a frameshift mutation, resulting in a product without the functional forkhead. These results have lead to speculation that the scurfy gene acts by disrupting the T-cell tolerance mechanism, resulting in hyperresponsiveness and lack of down-regulation. The Rag1KO/sf/Y OVA strain, with virtually 100% of its CD4+ T cells reactive strictly to ovalbumin (OVA) peptide 323-339, is an excellent model for determination of the sf mutation's ability to disrupt tolerance. We hypothesized that Rag1KO/sf/OVA mice would not be tolerant to antigen at a dose that tolerizes control animals. We found that splenic cells from Rag1KO/sf/Y OVA mice injected with the same dose of OVA peptide that induces tolerance in cells from control mice proliferate in vitro in response to OVA peptide. These results are consistent with a defect in the pathway responsible for peripheral T-cell tolerization. PMID- 11900417 TI - The Ergonomics Society--the Society Lectures 1999: the Ergonomics Society: 50 years of growth. AB - Developments in the Society are outlined since the early history was described by Edholm and Murrell. Major changes in the Society's operation and the context in which these changes took place are given. The changes in research directions, growth in educational facilities for professional education and the ever widening areas for the application of ergonomics are discussed. The consequences of these developments for the expansion of a recognition of the contribution of ergonomics, and the position of professionals in the international scene are touched upon. PMID- 11900415 TI - Early effects of tribromoethanol, ketamine/xylazine, pentobarbitol, and isoflurane anesthesia on hepatic and lymphoid tissue in ICR mice. AB - We investigated the effects of various anesthetic agents on hepatic and splenic injury in mice. Three and six hours after intraperitoneal injection of TBE, intramuscular injection of ketamine/xylazine combination (K/X), intraperitoneal injection of pentobarbital (PB), and inhalation of isoflurane (IF), or intraperitoneal and intramuscular injection of control saline, mice were exsanguinated and serum was obtained for measurement of hepatic aspartate transaminase (AST), alanine transaminase (ALT) and gamma-glutamyltransferase (GGT). The spleen and liver also were obtained, and sections were examined by use of routine light microscopy for pathologic changes and for apoptosis, as determined by use of the in situ terminal deoxynucleotidyl transferase-mediated dUPT nick-end-labeling (TUNEL) histochemical analysis. Three hours after TBE or K/X administration, AST activity increased three- to fourfold above that in untreated and saline-injected control animals, and remained high at six hours. Administration of PB did not effect AST activity at three hours, but there was a significant increase at six hours. Activity of ALT was non-significantly increased three hours after TBE and K/X, but not PB administration. Administration of IF had no effect on hepatic enzyme activities, and GGT was not increased after administration of any of the agents. Markedly increased apoptosis was observed in splenic follicles and in hepatic Kupffer and endothelial cells at three hours after TBE and K/X administration, but apoptosis decreased to control levels by six hours. Increased apoptosis was not observed after IF administration. Administration of TBE and K/X causes injury to lymphocytes and to hepatic Kupffer and endothelial cells within three hours, and PB administration induces changes within six hours. Thus, use of these anesthetic agents should be avoided when experiments are being designed to test short-term effects of an experimental intervention on the spleen and possibly on all lymphoid tissues. In addition, they also should be avoided in experiments testing effects on hepatic tissue. PMID- 11900416 TI - Intraocular teratoma in a mouse. AB - A 4.5-month-old male B6.129S2-Trp53tm1Tyj mouse developed an enlarged left eye. After euthanasia, the eye was removed and found to contain a teratoma composed of smooth muscle, white fat, neural tissue, and villous intestinal epithelium with appearance similar to that of the small intestine. Many normal intraocular structures were absent. To our knowledge, this represents the first reported intraocular teratoma in a mouse. PMID- 11900418 TI - The Ergonomics Society--the Society Lectures 1999: the next 50 years: future challenges and opportunities for empathy in our science. AB - To stay relevant and applicable in a rapidly changing world, ergonomics must meet several challenges. The paper explores these challenges and how the profession might respond. It is proposed that there are opportunities, and a need, to embrace empathy as a legitimate and useful tool. Empathy has value in three main areas. In research we can broaden our understanding of people and situations, learning 'why' as well as 'how and what' people do. Second, by identifying with the emotional as well as intellectual concerns of sponsors and colleagues from other professions, we inspire trust and confidence, adding value to our collective efforts in collaborative work. Third, through methods such as role playing and story telling, empathy is a powerful tool for getting ergonomics issues across to implementers and influencing outcomes. Care is needed to balance empathy with systematic observation in the tradition of the scientific method; the power of empathy is in combining it with objective methods to make full use of our abilities as human scientists. PMID- 11900419 TI - Unfocused ergonomics? AB - Ergonomics is now fairly routinely used in industry. Managers often initiate contact with ergonomists. The paper argues that many ergonomists have abandoned some fundamental ergonomics principles and have unwittingly succumbed to 'sitting complacently in the manager's toolbox'. It is also argued that through its inherent diversity, ergonomics is in danger of becoming fragmented and losing its identity. PMID- 11900420 TI - Another test of sensor lines on control panels. AB - In this experiment all panels had four circular displays arranged in a square on a vertical surface and four controls aligned vertically to the right of the displays. Two panels had linkages between controls and displays which earlier research had shown to be compatible, the other two had incompatible linkages. Each of these four panels was tested with and without sensor lines showing the linkages between controls and displays. Ninety-six subjects each made 128 trials on one of the eight panels. A subject's task was to respond as quickly as possible by pushing the correct control to extinguish a light when it appeared in one of the displays. Sensor lines had no effect on performance with the compatible panels but impaired performance on the incompatible panels. The results also confirm again the potency of the compatibility principle: responses were faster and fewer errors were made when using the compatible panels. PMID- 11900421 TI - Muscle activity and cardiovascular response during computer-mouse work with and without memory demands. AB - Computer-mouse work is characterized by repetitive movements combined with mental demands. The present purpose was to study how the body responded to simulated Computer Aided Design (CAD) work without memory demand and when a high short-term memory demand was introduced. Nine female subjects repetitively performed a task which involved 15 s of elevation of the right index and middle fingers followed by 6 s of rest. Every second time the fingers rested, the left index finger was required to type a six-figure number, either '123456' (without memory demand) or a random number shown half a minute before (with memory demand). After 7 min of performing the task without memory demand, the memory demand was introduced and continued for 1 h. Introduction of memory demand resulted in increased heart rate (77-->84 beats/min), blood pressure (systolic 129-->140 mmHg; diastolic 72-->79 mmHg) and forearm extensor muscle activity (wrist, 2.7-->4.5% EMGmax; finger, 5.6 ->7.5% EMGmax) and finger flexor muscle activity (0.7-->1.2% EMGmax) indicating increased co-contraction. Hereafter, muscle activity and cardiovascular response tended to decrease. Self-reported stress and rating of perceived exertion (RPE) for the right shoulder increased throughout the period. Two additional sub studies were inlcuded, which focused on adaptation to the physical load, showing a decrease in muscle activity and arousal, and reintroduction of the memory load, showing a lower response as compared to the initial response. The practical consequences of the findings suggest that job content should have variable mental demands. PMID- 11900422 TI - Falls and working individuals: role of extrinsic and intrinsic factors. AB - Falls are frequent occupational accidents involving workers and lead to important social and economic consequences both for the individual and for the employer. Different factors can modify balance control and lead to falling, especially environment-related and individual factors. The literature would appear to indicate that there have been few studies on the intrinsic factors involving the mechanisms of generating falls. This review determines the main factors involved in the mechanisms of falling, whether related to the environment, work or the individual. Knowledge of the extrinsic and intrinsic factors contributing to the fall could allow securer environment planning and occupational conditions for employers, and the use of balance rehabilitation methods for individuals to reduce the risk of falls. PMID- 11900423 TI - Investigation of membrane deformation by a fringe projection method. AB - An optical method has been developed to measure the deformation of a membrane in a microphone. A fringe projector that consists of an optical fiber and an optical wedge is described and analyzed by geometric optics. A three-step phase-shifting technique that involves the introduction of an arbitrary phase is discussed. The fine fringe patterns projected onto a small test surface are captured by a CCD camera mounted on a long-distance microscope. With the aid of a phase-shifting technique and signal-demodulating techniques, the proposed setup is capable of measuring deformation of the membrane of the order of as much as submicrometers. PMID- 11900424 TI - Correlation image velocimetry: a spectral approach. AB - A method, believed to be new, is introduced to evaluate displacement fields from the analysis of a deformed image compared with a reference image. In contrast to standard methods, which determine a piecewise constant displacement field, the present method gives direct access to spectral decomposition of the displacement field. A minimization procedure is derived and used twice: first, to determine an affine displacement field and, then, the spectral components of the residual displacement. Although the method is applicable to any space dimension, only cases dealing with one-dimensional signals are reported: First, a purely synthetic example is discussed to estimate the intrinsic performance of the method, and a second case deals with a profile extracted from a sample of compressed glass wool. PMID- 11900425 TI - Adaptive optics with four laser guide stars: correction of the cone effect in large telescopes. AB - We study the performance of an adaptive optics (AO) system with four laser guide stars (LGSs) and a natural guide star (NGS). The residual cone effect with four LGSs is obtained by a numerical simulation. This method allows the adaptive optics system to be extended toward the visible part of the spectrum without tomographic reconstruction of three-dimensional atmospheric perturbations, resolving the cone effect in the visible. Diffraction-limited images are obtained with 17-arc ms precision in median atmospheric conditions at wavelengths longer than 600 nm. The gain achievable with such a system operated on an existing AO system is studied. For comparison, performance in terms of achievable Strehl ratio is also computed for a reasonable system composed of a 40 x 40 Shack Hartmann wave-front sensor optimized for the I band. Typical errors of a NGS wave front are computed by use of analytical formulas. With the NGS errors and the cone effect, the Strehl ratio can reach 0.45 at 1.25 microm under good-seeing conditions with the Nasmyth Adaptive Optics System (NAOS; a 14 x 14 subpupil wave front sensor) at the Very Large Telescope and 0.8 with a 40 x 40 Shack-Hartmann wave-front sensor. PMID- 11900426 TI - Measurements of the corneal birefringence with a liquid-crystal imaging polariscope. AB - An imaging polariscope has been used to analyze the spatially resolved polarization properties of living human corneas. The apparatus is a modified double-pass setup, incorporating a liquid-crystal modulator in the analyzer pathway. Keeping the incident polarization state fixed (first passage), we recorded a series of three images of the pupil's plane corresponding to independent polarization states of the analyzer unit. Azimuth and retardation at each point of the cornea were calculated from those images. Results show that the magnitude of retardation increases along the radius toward the periphery of the cornea. Left-right eye symmetry in retardation was also found. Maps of azimuth indicate that the direction of the corneal slow axis is nasally downward. PMID- 11900427 TI - Laser profiling system for on-line measurement of superfine surfaces. AB - A profiling system for on-line and noncontact measurement of superfine surfaces is described. The system uses the principle of a differential interferometer with coaxial interference arms. The measuring head (optical part) of this system is separable and can be installed on machine tools or in existing measuring instruments. It has excellent resistance to vibration and air turbulence; therefore it needs no damper or cover. This system scans sample surfaces to measure a microprofile with a vertical resolution of 0.08 nm rms. PMID- 11900428 TI - Enhancement of the dynamic range of the detected intensity in an optical measurement system by a three-channel technique. AB - When three-dimensional optical topometry of technical surfaces is performed, one major problem is that often the local reflectance of the object's surface varies within a wide range. This leads to overexposed and underexposed areas on the detector, where no measurements can be made. To overcome this problem, we have developed a method that extends the dynamic range of an imaging system. As an example we implemented this method to a measurement system that is based on fringe projection with a data projector and a color CCD camera. By projection of quasi-monochromatic fringes a different dynamic range in each of the three color channels of the camera is achieved. Hence the overall dynamic range of the system is increased by a factor larger than 5. PMID- 11900429 TI - Numerical optimization of integrating cavities for diffraction-limited millimeter wave bolometer arrays. AB - Far-infrared to millimeter-wave bolometers designed to make astronomical observations are typically encased in integrating cavities at the termination of feedhorns or Winston cones. This photometer combination maximizes absorption of radiation, enables the absorber area to be minimized, and controls the directivity of absorption, thereby reducing susceptibility to stray light. In the next decade, arrays of hundreds of silicon nitride micromesh bolometers with planar architectures will be used in ground-based, suborbital, and orbital platforms for astronomy. The optimization of integrating cavity designs is required for achieving the highest possible sensitivity for these arrays. We report numerical simulations of the electromagnetic fields in integrating cavities with an infinite plane-parallel geometry formed by a solid reflecting backshort and the back surface of a feedhorn array block. Performance of this architecture for the bolometer array camera (Bolocam) for cosmology at a frequency of 214 GHz is investigated. We explore the sensitivity of absorption efficiency to absorber impedance and backshort location and the magnitude of leakage from cavities. The simulations are compared with experimental data from a room-temperature scale model and with the performance of Bolocam at a temperature of 300 mK. The main results of the simulations for Bolocam-type cavities are that (1) monochromatic absorptions as high as 95% are achievable with <1% cross talk between neighboring cavities, (2) the optimum absorber impedances are 400 ohms/sq, but with a broad maximum from approximately 150 to approximately 700 ohms/sq, and (3) maximum absorption is achieved with absorber diameters > or = 1.5 lambda. Good general agreement between the simulations and the experiments was found. PMID- 11900430 TI - Graded reflectivity micromirror arrays. AB - A technique for fabricating arrays of graded reflectivity micromirrors with diameters as small as 25 microm is reported. It is based on laser-induced physical vapor deposition through microholes on a thin free-standing noncontact mask, and it is suitable for applications in micro-optics and solid-state laser technology. PMID- 11900431 TI - Reflective light modulator based on epsilon-GaSe crystal. AB - We report the results of investigating a low-voltage, polarization-insensitive, reflective-type modulator based on an epsilon-GaSe crystal and operated at the 1.960-eV line of a He-Ne laser. We demonstrate that the modulation in an Al epsilon-GaSe-Cu device results mainly from the Franz-Keldysh effect. Relatively high speed and low operating voltage could make these modulators with Schottky barrier contacts attractive devices in the red range of the spectrum. PMID- 11900432 TI - Surface characterization techniques for determining the root-mean-square roughness and power spectral densities of optical components. AB - Surface topography and light scattering were measured on 15 samples ranging from those having smooth surfaces to others with ground surfaces. The measurement techniques included an atomic force microscope, mechanical and optical profilers, confocal laser scanning microscope, angle-resolved scattering, and total scattering. The samples included polished and ground fused silica, silicon carbide, sapphire, electroplated gold, and diamond-turned brass. The measurement instruments and techniques had different surface spatial wavelength band limits, so the measured roughnesses were not directly comparable. Two-dimensional power spectral density (PSD) functions were calculated from the digitized measurement data, and we obtained rms roughnesses by integrating areas under the PSD curves between fixed upper and lower band limits. In this way, roughnesses measured with different instruments and techniques could be directly compared. Although smaller differences between measurement techniques remained in the calculated roughnesses, these could be explained mostly by surface topographical features such as isolated particles that affected the instruments in different ways. PMID- 11900433 TI - Optical design of a stigmatic extreme-ultraviolet spectroscopic system for emission and absorption studies of laser-produced plasmas. AB - The design of a stigmatic spectroscopic system for diagnostics of laser-produced plasmas in the 2.5-40-nm region is presented. The system consists of a grazing incidence toroidal mirror that focuses the radiation emitted by a laser-produced plasma onto the entrance slit of a spectrograph. The latter has a grazing incidence spherical variable-line-spaced grating with flat-field properties coupled to a spherical focusing mirror that compensates for the astigmatism. The mirror is crossed with respect to the grating; i.e., it is mounted with its tangential plane coincident with the equatorial plane of the grating. The spectrum is acquired by an extreme-UV- (EUV-) enhanced CCD detector with high quantum efficiency. This stigmatic design also has spectral and spatial resolution capability for extended sources: The spectral resolution is also preserved for off-plane points, whereas the spatial resolution decreases for points far from the optical axis. The expected performance is presented and compared with that of a stigmatic design with a plane variable-line-spaced grating illuminated in converging light. PMID- 11900434 TI - Skin lesion classification using oblique-incidence diffuse reflectance spectroscopic imaging. AB - We discuss the use of a noninvasive in vivo optical technique, diffuse reflectance spectroscopic imaging with oblique incidence, to distinguish between benign and cancer-prone skin lesions. Various image features were examined to classify the images from lesions into benign and cancerous categories. Two groups of lesions were processed separately: Group 1 includes keratoses, warts versus carcinomas; and group 2 includes common nevi versus dysplastic nevi. A region search algorithm was developed to extract both one- and two-dimensional spectral information. A bootstrap-based Bayes classifier was used for classification. A computer-assisted tool was then devised to act as an electronic second opinion to the dermatologist. Our approach generated only one false-positive misclassification out of 23 cases collected for group 1 and two misclassifications out of 34 cases collected for group 2 under the worst estimation condition. PMID- 11900435 TI - Gravitational eccentric correction optics (GECO): an optical-gravitational device to compensate for flexures in astronomical spectrographs. AB - Mechanical flexure is a source of major failures in astronomical spectrographs, for which the reimaging of a focal-plane pinhole has to be maintained in position within a fraction of a CCD pixel that has dimensions of the order of 15 microm. The d.o.lo.res. (an acronym for device optimized for low resolution) spectrograph for the Italian national telescope, Galileo, showed displacements of the image of the pinhole more than 10 times greater than expected. The mechanical failure was overcome by the insertion of a passive optical wedge that can add an out-of-phase circle to the flexure ellipse. The results encourage the use of the gravitational eccentric correction optics (GECO) optical-gravitational device in all astronomical observations made with the d.o.lo.res. spectrograph. PMID- 11900436 TI - Electro-optically modulated polarizing Fourier-transform spectrometer for plasma spectroscopy applications. AB - A new electro-optically modulated optical solid-state (MOSS) interferometer has been constructed for measurement of quantities related to the low-order spectral moments of line emission from optically thin radiant media such as plasmas. When Doppler broadening is dominant, the spectral moments give the Radon transform of corresponding moments of the velocity distribution function of the radiating species. The instrument, which is based on the principle of the Fourier-transform spectrometer, has high etendue and is rugged and compact. When electro-optical path-length modulation techniques are employed, the spectral information is encoded in the temporal frequency domain at harmonics of the modulation frequency and can be obtained by use of a single photodetector. Specifically, for a plasma in drifting local thermodynamic equilibrium the zeroth moment (brightness) is given by the average signal level, the first moment (shift) by the interferometric phase, and the second moment (linewidth) by the fringe visibility. To illustrate the MOSS performance, I present spectroscopic measurements of the time evolution of the plasma ion temperature and flow velocity for rf-heated discharges in the H-1 heliac, a toroidal plasma magnetic confinement at the Australian National University. PMID- 11900437 TI - Spectral characterization of acousto-optic filters used in imaging spectroscopy. AB - The purpose of this investigation is to improve the study of the characteristics of noncollinear acoustooptic tunable filters (AOTFs) used in imaging spectroscopy. Three filters were characterized and the results compared with tuning models to verify that device operation can be reliably predicted in advance. All these devices use tellurium dioxide as the interaction medium and have large geometric apertures for spectroscopic imaging applications in the spectral range 0.5-3.5 microm. The device characteristics that we studied were compared with the results of AOTF models, and the spectral and angular dependence of acoustic frequency and bandpass width for both output polarization states were confirmed by measurements. One of the AOTFs was used as a dispersive element coupled to external imaging optics. We summarize measurements of the basic spectral and imaging characteristics in this configuration. PMID- 11900438 TI - Measurement method for profiling the residual stress and the strain-optic coefficient of an optical fiber. AB - A method, believed novel, is demonstrated for determining the strain-optic coefficient profile as well as the residual-stress profile of an optical fiber by use of a modified polariscope combined with a fiber-elongation apparatus. Measurement results of the residual-stress and the strain-optic coefficient profiles for Ge-doped and Er-Ge-Al-doped optical fibers are demonstrated with this method. PMID- 11900439 TI - Multiscale mapping technique for the simultaneous estimation of absorption and partial scattering in optical coatings. AB - The control and the characterization of optical losses are crucial elements in the design of high-quality thin films. Nonuniformity of losses and the existence of local defects have led us to perform simultaneous absorption and scattering mapping in exactly the same experimental conditions. An improved setup and new procedures are capable of providing such paired mappings of absorption and partial scattering at various spatial scales. The diameter of the pump beam, which governs lateral spatial resolution, can be chosen to be 3-100 microm. The detectivity threshold can be as low as 0.1 part in 10(6) for absorption and 0.01 part in 10(6) for mapping partial scattering. Spatial windows can range from micrometer-sized areas for the study of micro defects to centimeter-sized areas on which the uniformity of losses can be checked. We study the spatial distribution of absorption and scattering losses under scale transformation by changing the spatial window while keeping the spatial resolution constant. We present one-dimensional and bidimensional multiscale studies. For example, we show that one can use multiscale mapping of defects to evaluate the qualities of substrate cleaning, which are not identical on micrometric and centimetric scales. PMID- 11900440 TI - X-ray multilayer monochromator with enhanced performance. AB - An x-ray multilayer monochromator with improved resolution and a low specular background is presented. The monochromator consists of a lamellar multilayer amplitude grating with appropriate parameters used at the zeroth diffraction order. The device is fabricated by means of combining deposition of thin films on a nanometer scale, UV lithography, and reactive ion etching. The performance of this new monochromator at photon energies near 1500 eV is shown. PMID- 11900441 TI - Grazing-incidence Monk-Gillieson monochromator based on surface normal rotation of a varied-line-spacing grating. AB - A geometric theory of a grazing-incidence varied-line-spacing plane-grating monochromator system whose scanning is made by a simple grating rotation about the grating normal has been developed for designing Monk-Gillieson monochromators capable of covering an energy range of 0.6-2.5 keV. Analytic expressions are given for the grating equations, focal conditions, dispersion, spectral image shape, and optimization of groove parameters. On the basis of the theory, two monochromator systems have been designed: system I for moderate resolution and system II for relatively high resolution. The validity of the analytic formulas and the expected performance of the designed systems have been evaluated by means of ray tracing. The results show that the analytic formulas are sufficiently accurate for practical applications and that systems I and II would provide resolving power of approximately 1450-600 and 7500-2000, respectively, in the wavelength region of 0.5-2.0 nm. PMID- 11900442 TI - Dynamic optical coherence tomography in studies of optical clearing, sedimentation, and aggregation of immersed blood. AB - The concept of refractive-index matching to enhance the optical penetration depth of whole blood is discussed on the basis of in vitro studies that used the technique of near-infrared optical coherence tomography. It was found that optical clearing of blood is defined not only by refractive-index matching but also by changes in the size of red blood cells and in their aggregation ability when chemicals are added. For example, in whole blood diluted to twice its volume by saline with the addition of 6.5% glycerol, the total attenuation coefficient was reduced from 4.2 to 2.0 mm(-1), and the optical penetration at 820 nm was correspondingly increased to 117%. For the other agents tested (glucose, dextrans, propylene glycol, and trazograph) the enhancement of penetration was 20 150.5%. In the blood sedimentation study, regular or irregular oscillations or jumps of the red-blood cell-plasma boundary were observed. The 1-min time period of regular oscillations correlated well with the kinetics of the aggregation process as described by the two subsequent stages of formation of linear and three-dimensional aggregates. The results also showed that optical clearing of blood by osmotic agents is potentially useful not only in blood sedimentation and aggregation studies but also in intravascular optical coherence tomography imaging techniques. PMID- 11900443 TI - Polarization imaging by use of digital holography. AB - We present what we believe to be a new digital holographic imaging method that is able to determine simultaneously the distributions of intensity, phase, and polarization state at the surface of a specimen on the basis of a single image acquisition. Two reference waves with orthogonal polarization states interfere with the object wave to create a hologram that is recorded on a CCD camera. Two wave fronts, one for each perpendicular polarization state, are numerically reconstructed in intensity and phase. Combining the intensity and the phase distributions of these two wave fronts permits the determination of all the components of the Jones vector of the object-wave front. We show that this method can be used to image and measure the distribution of the polarization state at the surface of a specimen, and the obtained results indicate that precise quantitative measurements of the polarization state can be achieved. An application of the method to image the birefringence of a stressed polymethyl methacrylate sample is presented. PMID- 11900444 TI - Differential interference contrast microscopy as a polarimetric instrument. AB - Differential interference contrast (DIC) microscopy is shown to be equivalent to an incomplete Stokes polarimeter capable of probing optical properties of materials on microscopic-length scales. The Mueller matrix for a DIC microscope is calculated for various types of samples, and the polarimetric properties for DIC component parts of a spaceflight microscope are spectrally measured. As a practical application, a measurement of the index mismatch between colloidal particles and a nearly index-matched fluid bath was performed. PMID- 11900445 TI - Phase-shifting algorithms for electronic speckle pattern interferometry. AB - A set of innovative phase-shifting algorithms developed to facilitate metrology based on electronic speckle pattern interferometry (ESPI) are presented. The theory of a phase-shifting algorithm, called a (5,1) algorithm, that takes five phase-shifted intensity maps before a specimen is deformed and one intensity map after a specimen is deformed is presented first. Because a high-speed camera can be used to record the dynamic image of the specimen, this newly developed algorithm has the potential to retain the phase-shifting capability for ESPI in dynamic measurements. Also shown is an algorithm called a (1,5) algorithm that takes five phase-shifted intensity maps after the specimen is deformed. In addition, a direct-correlation algorithm was integrated with these newly developed (5,1) or (1,5) algorithms to form DC-(5,1) and DC-(1,5) algorithms, which are shown to improve significantly the quality of the phase maps. The theoretical and experimental aspects of these two newly developed techniques, which can extend ESPI to areas such as high-speed dynamic measurements, are examined in detail. PMID- 11900446 TI - Phase-stepping interferometry: methods for reducing errors caused by camera nonlinearities. AB - Phase errors that arise in phase-stepping interferometry are discussed. Investigations were performed by use of a Twyman-Green interferometer equipped with a compensation plate with a variable and servo-controlled tilt angle. With this instrument, phase-stepping errors can be reduced to a negligible level. There are, however, phase errors that are caused by camera nonlinearities. Two methods for minimizing these errors are presented. The first method is based on the simple idea that the interference intensity at the output of a two-beam interferometer has an exact cosine shape. The camera signals were monitored as a function of the tilt angle of the compensation plate, and the deviation from the cosine form was used to produce a correction. The second method is based on the idea that, under specific conditions, errors of an average of two phase measurements may compensate for each other. Numerical calculations were performed and give evidence of this hypothesis. Each method, the signal-correction and the averaging method, drastically reduces errors in evaluation of phases. The combination of both methods is a powerful tool that allows precise phase data to be obtained with an uncertainty, in the range lambda/2000 approximately 0.3 nm, that is caused mainly by signal noise. PMID- 11900447 TI - Form assessment of hollow cylindrical specimens. AB - Grazing-incidence interferometry that makes use of diffractive axicons for the measurement of cylindrical mantle surfaces has already been reported. However, measurement of concave rod structures poses a severe problem because these structures are subject to spurious fringes caused by parasitic diffraction orders of the diffractive axicons. By breaking the symmetry of the interferometric setup it is possible to obtain unique interferograms of the inner mantle surfaces of hollow cylinders as cages for roller bearings or other workpieces produced on lathe machines that have a suitable surface finish. Special design issues for the computer-generated holograms and the interferometric setup are discussed, and test examples are given. PMID- 11900448 TI - Design and analysis of an integrated antiresonant reflecting optical waveguide refractive-index sensor. AB - An integrated optical waveguide refractometer, believed to be novel, is presented. The sensor is based on an antiresonant reflecting optical waveguide and uses the strong attenuation dependence on the refractive index of antiresonant cladding layers as the sensing principle. The theory and the operation of the sensor are discussed in terms of one- and two-dimensional geometry. The theoretical predictions and numerical analysis show that a versatile sensor can be realized. The design trade-offs are discussed, and the sensitivity and measurement range are presented. PMID- 11900449 TI - Polarization dependence in Ti-diffused LiNbO3 directional couplers. AB - The effect of design and fabrication parameters on the dependence on polarization of the splitting ratio in directional couplers produced by Ti diffusion in LiNbO3 has been investigated experimentally at a wavelength of 1.54 microm. The parametric study was carried out at a diffusion temperature of 1025 degrees C, which was found to suppress outdiffusion effects. The directional couplers were produced by use of various combinations of waveguide separation, Ti film thickness, and diffusion time. Of particular interest was the identification of parameter sets for which the sum of TE and TM splitting ratios equals unity, as required for electro-optic and acousto-optic tunable filter designs with relaxed beam-splitter requirements. Directional couplers that closely match this criterion were obtained by diffusion of 3.5-mm-long, 7-microm-wide Ti strips separated by 11 microm for 11 h. It was found that a bending angle of 0.6 degrees for input and output waveguides produces lower transmission loss for both polarizations than a 1.0 degree bend angle (>1-dB loss reduction). PMID- 11900450 TI - Weigert's effect mechanism observed in dyes. AB - Anisotropic grains have been shown to be spontaneously induced in films that have been oversaturated and overcooled with dye in which Weigert's effect can be observed. Some of the grains exposed to active linearly polarized light are destroyed with the result that the film becomes anisotropic. PMID- 11900451 TI - Spatially modulated illumination microscopy allows axial distance resolution in the nanometer range. AB - For an improved understanding of the structural basis of cellular mechanisms, it is highly desirable to develop methods for a detailed topological analysis of biological nanostructures and their dynamics in the interior of three dimensionally conserved cells. We present a method of far-field laser fluorescence microscopy to measure relative axial positions of pointlike fluorescent targets and the distance between each target in the range of a few nanometers. The physical principle behind this approach can be extended to the determination of three-dimensional (3D) positions and 3D distances between any number of objects that can be discriminated owing to their spectral signature, thus allowing topological measurements so far regarded to be beyond the capabilities of light microscopy. PMID- 11900452 TI - Effect of polishing conditions on terminating optical connectors with spherical convex polished ends. AB - Increased demand for fiber-optic technology has created significant growth in the sales of interconnection devices such as fiber-optic connectors, cable assemblies, and adapters. To ensure good connector performance during actual use, several process parameters related to geometric and optical characteristics of the connector must be thoroughly understood during the manufacturing stage. The experimental design has been used here to see the influence of applied pressure and time on the fiber end geometry as well as optical performance. The mathematical model is also applied to explain the phenomena of the present fiber undercut-reflectance relation. By a proper choice of polishing film grit size and processing conditions, it is possible to obtain fiber connectors with less fiber undercut and better return loss. Influences of film grit size and rubber-pad thickness on the reflectance and the fiber undercut are also presented. PMID- 11900453 TI - Polarization-multiplexed diffractive optical elements fabricated by subwavelength structures. AB - Polarization-multiplexed phase-only diffractive optical elements with subwavelength structures are proposed and fabricated. The differences among the phase modulations result from the differences among the effective indices exhibited in the subwavelength structures with various filling factors and surface profiles, and the phase retardations are obtained by the relief depth of the structures. The polarization-selective property is achieved by the polarization dependence of the effective indices exhibited in the one-dimensional subwavelength structures and the polarization independence exhibited in the two dimensional structures. Additionally, the polarization contrast of our polarization-multiplexed elements, defined as the cross talk between the two polarization incidences, is independent of the relief depth. The principle of the polarization multiplexing by use of the subwavelength structures is described, and the fabrication results for the polarization-multiplexed computer-generated holograms are demonstrated. PMID- 11900454 TI - TCF transcription factors, mediators of Wnt-signaling in development and cancer. PMID- 11900456 TI - Functional comparison of the Hoxa 4, Hoxa 10, and Hoxa 11 homeoboxes. AB - A number of models attempt to explain the functional relationships of Hox genes. The functional equivalence model states that mammalian Hox-encoded proteins are largely functionally equivalent, and that Hox quantity is more important than Hox quality. In this report, we describe the results of two homeobox swaps. In one case, the homeobox of Hoxa 11 was replaced with that of the very closely related Hoxa 10. Developmental function was assayed by analyzing the phenotypes of all possible allele combinations, including the swapped allele, and null alleles for Hoxa 11 and Hoxd 11. This chimeric gene provided wild-type function in the development of the axial skeleton and male reproductive tract, but served as a hypomorph allele in the development of the appendicular skeleton, kidneys, and female reproductive tract. In the other case, the Hoxa 11 homeobox was replaced with that of the divergent Hoxa 4 gene. This chimeric gene provided near recessive null function in all tissues except the axial skeleton, which developed normally. These results demonstrate that even the most conserved regions of Hox genes, the homeoboxes, are not functionally interchangeable in the development of most tissues. In some cases, developmental function tracked with the homeobox, as previously seen in simpler organisms. Homeoboxes with more 5' cluster positions were generally dominant over more 3' homeoboxes, consistent with phenotypic suppression seen in Drosophila. Surprisingly, however, all Hox homeoboxes tested did appear functionally equivalent in the formation of the axial skeleton. The determination of segment identity is one of the most evolutionarily ancient functions of Hox genes. It is interesting that Hox homeoboxes are interchangeable in this process, but are functionally distinct in other aspects of development. PMID- 11900455 TI - Inhibition of BMP activity by the FGF signal promotes posterior neural development in zebrafish. AB - The expression patterns of region-specific neuroectodermal genes and fate-map analyses in zebrafish gastrulae suggest that posterior neural development is initiated by nonaxial signals, distinct from organizer-derived secreted bone morphogenetic protein (BMP) antagonists. This notion is further supported by the misexpression of a constitutively active form of zebrafish BMP type IA receptor (CA-BRIA) in the zebrafish embryos. It effectively suppressed the anterior neural marker, otx2, but not the posterior marker, hoxb1b. Furthermore, we demonstrated that the cells in the presumptive posterior neural region lose their neural fate only when CA-BRIA and Xenopus dominant-negative fibroblast growth factor (FGF) receptors (XFD) are coexpressed. The indications are that FGF signaling is involved in the formation of the posterior neural region, counteracting the BMP signaling pathway within the target cells. We then examined the functions of Fgf3 in posterior neural development. Zebrafish fgf3 is expressed in the correct place (dorsolateral margin) and at the correct time (late blastula to early gastrula stages), the same point that the most precocious posterior neural marker, hoxb1b, is first activated. Unlike other members of the FGF family, Fgf3 had little mesoderm-inducing activity. When ectopically expressed, Fgf3 expands the neural region with suppression of anterior neural fate. However, this effect was mediated by Chordino (zebrafish Chordin), because Fgf3 induces chordino expression in the epiblast and Fgf3-induced neural expansion was substantially suppressed in dino mutants with mutated chordino genes. The results obtained in the present study reveal multiple actions of the FGF signal on neural development: it antagonizes BMP signaling within posterior neural cells, induces the expression of secreted BMP antagonists, and suppresses anterior neural fate. PMID- 11900457 TI - The second meiosis occurs in cytochalasin D-treated eggs of Corbicula leana even though it is not observed in control androgenetic eggs because the maternal chromosomes and centrosomes are extruded at first meiosis. AB - The hermaphroditic freshwater clam Corbicula leana reproduces by androgenesis. In the control (androgenetic development), all maternal chromosomes and maternal centrosomes at the meiotic poles were extruded as the two first polar bodies, and subsequently, second meiosis did not occur. But, in C. leana eggs treated with cytochalasin D (CD) to inhibit polar body extrusion, the second meiosis occurred. At metaphase-I, the spindle showed the typical bipolar structure and two spheroid centrosomes were located at its poles. All the maternal chromosomes were divided at anaphase-I, but they were not extruded as polar bodies due to the effects of CD. After completion of first meiosis, the maternal centrosomes split into four. At the second meiosis, twin or tetrapolar spindles were formed and two groups of maternal chromosomes divided into four sets of chromosomes. After the second meiosis, the spindle disassociated and the four maternal centrosomes disappeared. Four groups of maternal chromosomes transformed into the four female pronuclei. Male and female pronuclei became metaphase chromosomes of the first mitosis. The present study clearly indicates that typical meiosis systems still proceed in androgenetic triploid C. leana. We conclude that the androgenetic form may have arisen from the meiotic form. PMID- 11900458 TI - Endogenous patterns of BMP signaling during early chick development. AB - Bone morphogenetic proteins (BMPs) are members of the transforming growth factor beta superfamily signaling molecules that play important roles in a wide variety of developmental processes. In this study, we have used an antibody specific for the phosphorylated and activated form of Smad1 to examine endogenous patterns of BMP signaling in chick embryos during early development. We find complex spatial and temporal distributions of BMP signaling that elucidate how BMPs may function in multiple patterning events in the early chick embryo. In the pregastrula embryo, we find that BMP signaling is initially ubiquitous and is extinguished in the epiblast at the onset of primitive streak formation. At the head process stage, BMP signaling is inactivated in prospective neural plate, while it is strongly activated at the neural plate border, a region which is populated by cells that will give rise to neural crest. During later development, we find a dynamic spatiotemporal activation of BMP signaling along the rostrocaudal axis, in the dorsal neural tube, in the notochord, and in the somites during their maturation process. We discuss the implication of our results for endogenous functions of BMP signaling during chick development. PMID- 11900459 TI - Regional specification of the head and trunk-tail organizers of a urodele (Cynops pyrrhogaster) embryo is patterned during gastrulation. AB - The dorsal marginal zone (DMZ) of an amphibian early gastrula is thought to consist of at least two distinct domains: the future head and trunk-tail organizers. We studied the mechanism by which the organizing activities of the lower half of the DMZ (LDMZ) of the urodelean (Cynops pyrrhogaster) embryo are changed. The uninvoluted LDMZ induces the notochord and then organizes the trunk tail structures, whereas after cultivation in vitro or suramin treatment, the same LDMZ loses the notochord-inducing ability and organizes the head structures. A cell-lineage experiment indicated that the change in the organizing activity of the LDMZ was reflected in the transformation of the inductive ability: from notochord-inducing to neural-inducing activity. Using RT-PCR, we showed that the LDMZ expressed gsc, lim-1, chordin, and noggin, but not the mesoderm marker bra. In the sandwich assay, the LDMZ induced bra expression in the animal cap ectoderm, but the inductive activity was inhibited by cultivation or suramin treatment. The present study indicates that the change in the organizing activity of the LDMZ from trunk-tail to head is coupled with the loss of notochord inducing activity. Based on these results, we suggest that this change is essential for the specification of the head and trunk-tail organizers during gastrulation. PMID- 11900460 TI - Sonic hedgehog is required early in pancreatic islet development. AB - Pancreatic organogenesis relies on a complex interplay of cell-autonomous and extracellular signals. We demonstrate that the morphogen sonic hedgehog (Shh) is required for pancreatic development in zebrafish. Genetic mutants of Shh and its signaling pathway establish this dependence as specific to endocrine, but not exocrine, pancreas. Using cyclopamine to inhibit hedgehog signaling, we show that transient Shh signaling is necessary during gastrulation for subsequent differentiation of endoderm into islet tissue. A second hedgehog-dependent activity occurring later in development was also identified and may be analogous to the known action of Shh in gut endoderm to direct localization of pancreatic development. The early action of Shh may be part of a more general process allowing neuroendocrine cells to originate in nonneuroectodermally derived tissues. PMID- 11900461 TI - In vitro development of growing oocytes from fetal mouse oocytes: stage-specific regulation by stem cell factor and granulosa cells. AB - The development of follicles in the mammalian ovary involves a bidirectional communication system between the follicular cells and oocyte that is now beginning to be characterized. Little is known about the mechanisms underlying the beginning of the oocyte growth and the acquisition of the competence to resume meiosis by the growing oocyte. In the present study, we devised a multistep culture system for mouse oocytes obtained from 15.5- to 16.5-days postcoitum embryos (mean diameter +/- SEM, 9.7 +/- 1.3 microm), allowing three stages of the oocyte growth to be identified: (i) an early stage in which the oocyte growth is induced by direct stimulation of a soluble growth factor, namely stem cell factor (SCF), independent of the formation of gap junctions with granulosa cells; (ii) a second phase in which the oocyte growth depends on the combined action of SCF and contacts with granulosa cells; and (iii) a third phase of granulosa cell-dependent, SCF-independent growth. At each stage, key events of oocyte development and differentiation, such as the c-kit reexpression, the early zona pellucida assembly, and the beginning of follicologenesis, were observed to occur independently by the presence of SCF. At the end of the in vitro growing phases, lasting 18-20 days, oocytes reached a size (50 +/- 2.5 microm) and a chromatin differentiation (stage I-II) equivalent to those of 9- to 10-day-old preantral oocytes and were unable to complete the growth phase. About 50% of the in vitro-grown oocytes were induced to resume meiosis by okadaic acid (OA) treatment. However, a significant fraction of them (48%) showed inability to maintain the chromosome condensation in M-phase. When in vitro-grown oocytes were treated with UO126, a specific MEK inhibitor that prevents activation of mitogen activated protein kinases (ERK-1 and ERK-2), for 1 h before, during, and following OA treatment, only 22% of oocytes underwent germinal vesicle breakdown after 24 h from the OA treatment. These studies demonstrate that SCF alone can induce the onset of the oocyte growth. This is, however, not sufficient to fully activate the mechanisms governing the acquisition of the meiotic competence previously described as a 15-day oocyte-autonomous clock starting at the onset of growth. The inability of oocytes to progress into the last stages of growth and the lack of synchrony between nuclear and cytoplasm maturation showed by a subset of them resemble the characteristics of oocytes from connexin-37- and -43 deficient mice and indicate the preantral/antral transition point as a critical stage of oocyte development requiring the coordinated differentiation of the oocyte with granulosa cells and the maintenance of adequate communication between these two cell types to assure the correct oocyte meiotic maturation. PMID- 11900462 TI - Cooperation of Hoxa5 and Pax1 genes during formation of the pectoral girdle. AB - Hox and Pax transcription factors are master regulators of skeletal and organ morphogenesis. Some skeletal malformations encountered in Hoxa5 mutants are shared by the undulated (un) mice, which bear a point mutation in the Pax1 gene. To investigate whether Hoxa5 and Pax1 act in common pathways during skeletal development, we analyzed Hoxa5;un compound mutants. Our genetic studies show that Hoxa5 and Pax1 cooperate in the vertebral patterning of the cervicothoracic transition region and in acromion morphogenesis. The dynamics of expression of Hoxa5 and Pax1 in the pectoral girdle region suggest that both genes function in a complementary fashion during acromion formation. Whereas Pax1 is required for the recruitment of acromion precursor cells, Hoxa5 may provide regional cues essential for the correct formation of the acromion by ensuring Pax1 expression at the proper time and position during morphogenesis of the pectoral girdle. Hoxa5 also has a distinctive role in specifying the fate of perichondrial and chondrogenic cell lineages in a Sox9-dependent way. PMID- 11900463 TI - Enhanced branching morphogenesis in mammary glands of mice lacking cell surface beta1,4-galactosyltransferase. AB - Development of the mammary gland is influenced both by the systemic hormonal environment and locally through cell-cell and cell-extracellular matrix (ECM) interactions. We have previously demonstrated aberrant mammary gland morphogenesis in transgenic mice with elevated levels of the long isoform of beta1,4-galactosyltransferase 1 (GalT), a proportion of which is targeted to the plasma membrane, where it plays a role in cell-ECM interactions. Here, we show that mammary glands of mice lacking the long GalT isoform exhibit a complementary phenotype. Cell-surface GalT activity was reduced by over 60%, but because the short GalT isoform is intact, total GalT activity was reduced only slightly relative to wild type. Mammary glands from long GalT-null mice were characterized by excess branching, and this phenotype was accompanied by altered expression of laminin chains. Laminin alpha1 and alpha3 were reduced 2.4- and 3.0-fold, respectively, while expression of laminin gamma2 was elevated 2.3-fold. The expression and cleavage of laminin gamma2 have been correlated with branching and cell migration, and Western blotting revealed an altered pattern in gamma2 cleavage products in long GalT-null mammary glands. We then examined the expression of metalloproteases that cleave laminins or that have been shown to play a role in mammary gland morphogenesis. Expression of MT1-MMP, a membrane bound protease that can cleave laminin gamma2, was elevated 5.5-fold in the long GalT-nulls. MMP 7 was also elevated 5.1-fold. Our results suggest that expression of surface GalT is important for the proper regulation of matrix expression and deposition, which in turn regulates the proper branching morphogenesis of the mammary epithelial ductal system. PMID- 11900465 TI - Tangential migration in neocortical development. AB - During cortical development, different cell populations arise in the basal telencephalon and subsequently migrate tangentially to the neocortex. However, it is not clear whether these cortical cells are generated in the lateral ganglionic eminence (LGE), the medial ganglionic eminence (MGE), or both. In this study, we have generated a three-dimensional reconstruction to study the morphological formation of the two ganglionic eminences and the interganglionic sulcus. As a result, we have demonstrated the importance of the development of these structures for this tangential migration to the neocortex. We have also used the tracers DiI and BDA in multiple experimental paradigms (whole embryo culture, in utero injections, and brain slice cultures) to analyze the routes of cell migration and to demonstrate the roles of both eminences in the development of the cerebral cortex. These results are further strengthened, confirming the importance of the MGE in this migration and demonstrating the early generation of tangential migratory cells in the LGE early in development. Finally, we show that the calcium-binding protein Calretinin is expressed in some of these tangentially migrating cells. Moreover, we describe the spatiotemporal sequence of GABA, Calbindin, and Calretinin expression, showing that these three markers are expressed in the cortical neuroepithelium over several embryonic days, suggesting that the cells migrating tangentially form a heterogeneous population. PMID- 11900464 TI - Different isoforms of fasciclin II are expressed by a subset of developing olfactory receptor neurons and by olfactory-nerve glial cells during formation of glomeruli in the moth Manduca sexta. AB - During development of the primary olfactory projection, olfactory receptor axons must sort by odor specificity and seek particular sites in the brain in which to create odor-specific glomeruli. In the moth Manduca sexta, we showed previously that fasciclin II, a cell adhesion molecule in the immunoglobulin superfamily, is expressed by the axons of a subset of olfactory receptor neurons during development and that, in a specialized glia-rich "sorting zone," these axons segregate from nonfasciclin II-expressing axons before entering the neuropil of the glomerular layer. The segregation into fasciclin II-positive fascicles is dependent on the presence of the glial cells in the sorting zone. Here, we explore the expression patterns for different isoforms of Manduca fasciclin II in the developing olfactory system. We find that olfactory receptor axons express transmembrane fasciclin II during the period of axonal ingrowth and glomerulus development. Fascicles of TM-fasciclin II+ axons target certain glomeruli and avoid others, such as the sexually dimorphic glomeruli. These results suggest that TM-fasciclin II may play a role in the sorting and guidance of the axons. GPI-linked forms of fasciclin II are expressed weakly by glial cells associated with the receptor axons before they reach the sorting zone, but not by sorting zone glia. GPI-fasciclin II may, therefore, be involved in axon-glia interactions related to stabilization of axons in the nerve, but probably not related to sorting. PMID- 11900466 TI - The expression of the let-7 small regulatory RNA is controlled by ecdysone during metamorphosis in Drosophila melanogaster. AB - In Caenorhabditis elegans, the heterochronic pathway controls the timing of developmental events during the larval stages. A component of this pathway, the let-7 small regulatory RNA, is expressed at the late stages of development and promotes the transition from larval to adult (L/A) stages. The stage-specificity of let-7 expression, which is crucial for the proper timing of the worm L/A transition, is conserved in Drosophila melanogaster and other invertebrates. In Drosophila, pulses of the steroid hormone 20-hydroxyecdysone (ecdysone) control the timing of the transition from larval to pupal to adult stages. To test whether let-7 expression is regulated by ecdysone in Drosophila, we used Northern blot analysis to examine the effect of altered ecdysone levels on let-7 expression in mutant animals, organ cultures, and S2 cultured cells. Experiments were conducted to test the role of Broad-Complex (BR-C), an essential component in the ecdysone pathway, in let-7 expression. We show that ecdysone and BR-C are required for let-7 expression, indicating that the ecdysone pathway regulates the temporal expression of let-7 in Drosophila. These results demonstrate an interaction between steroid hormone signaling and the heterochronic pathway in insects. PMID- 11900467 TI - Evidence for a cell-specific action of Reelin in the spinal cord. AB - Reelin, the extracellular matrix protein missing in reeler mice, plays an important role in neuronal migration in the central nervous system. We examined the migratory pathways of phenotypically identified spinal cord neurons to determine whether their positions were altered in reeler mutants. Interneurons and projection neurons containing choline acetyltransferase and/or NADPH diaphorase were studied in E12.5-E17.5 reeler and wild-type embryos, and their final locations were assessed postnatally. While three groups of dorsal horn interneurons migrated and differentiated normally in reeler mice, the migrations of both sympathetic (SPNs) and parasympathetic preganglionic neurons (PPNs) were aberrant in the mutants. Initially reeler and wild-type SPNs were detected laterally near somatic motor neurons, but by E13.5, many reeler SPNs had mismigrated medially. Postnatally, 79% of wild-type SPNs were found laterally, whereas in reeler, 92% of these neurons were positioned medially. At E13.5, both reeler and wild-type PPNs were found laterally, but by E14.5, reeler PPNs were scattered across the intermediate spinal cord while wild-type neurons correctly maintained their lateral location. By postnatal day 16, 97% of PPNs were positioned laterally in wild-type mice; in contrast, only 62% of PPNs were found laterally in mutant mice. In E12.5-E14.5 wild-type mice, Reelin-secreting cells were localized along the dorsal and medial borders of both groups of preganglionic neurons, but did not form a solid barrier. In contrast, Dab1, the intracellular adaptor protein thought to function in Reelin signaling, was expressed in cells having positions consistent with their identification as SPNs and PPNs. In combination, these findings suggest that, in the absence of Reelin, both groups of autonomic motor neurons migrate medially past their normal locations, while somatic motor neurons and cholinergic interneurons in thoracic and sacral segments are positioned normally. These results suggest that Reelin acts in a cell-specific manner on the migration of cholinergic spinal cord neurons. PMID- 11900468 TI - Developmental changes in Notch1 and numb expression mediated by local cell-cell interactions underlie progressively increasing delta sensitivity in neural crest stem cells. AB - Neural stem cells become progressively less neurogenic and more gliogenic with development. Here, we show that between E10.5 and E14.5, neural crest stem cells (NCSCs) become increasingly sensitive to the Notch ligand Delta-Fc, a progliogenic and anti-neurogenic signal. This transition is correlated with a 20- to 30-fold increase in the relative ratio of expression of Notch and Numb (a putative inhibitor of Notch signaling). Misexpression experiments suggest that these changes contribute causally to increased Delta sensitivity. Moreover, such changes can occur in NCSCs cultured at clonal density in the absence of other cell types. However, they require local cell-cell interactions within developing clones. Delta-Fc mimics the effect of such cell-cell interactions to increase Notch and decrease Numb expression in isolated NCSCs. Thus, Delta-mediated feedback interactions between NCSCs, coupled with positive feedback control of Notch sensitivity within individual cells, may underlie developmental changes in the ligand-sensitivity of these cells. PMID- 11900470 TI - Neurolytic blockade of the obturator nerve for intractable spasticity of adductor thigh muscles. AB - Neurolytic blockade is one of the therapeutic possibilities to treat spasticity of various muscles. In patients with spasticity of the adductor thigh muscles, a percutaneous approach to the obturator nerve is often difficult. We describe a new approach to the obturator nerve and we examine its feasibility. The second objective was to assess the efficacy of obturator neurolysis for the management of adductor thigh muscle pain and spasticity associated with hemiplegia or paraplegia. Nerve blocks were performed via a combined approach using fluoroscopy and nerve stimulation to identify the obturator nerve. Neurolysis was performed by injection of 65% ethanol. We performed 27 blocks in 23 patients. Technical evaluation was achieved in terms of number of attempted needle insertions, time to accurate location of the nerve and success rate. The efficacy of the block was assessed using four scores: degree of alleviation of muscle spasm and triple flexion of the lower limb, improvement of gait and facilitation of hygienic care. Success rate of the technique was 100% with a time to accurate nerve location of 130+/-35 s. Compared with scores measured immediately before the block, all studied parameters were significantly improved. Efficiency was significant on adductor muscle spasticity (p<0.001 at 1 day and p<0.01 at 60 and 120 months). Triple flexion was also significantly improved (p<0.05 from 1 to 120 days), as well as gait (p<0.02) and hygiene (p<0.01) scores. No complications occurred. The combined approach of the obturator nerve represents a new technique which proved to be accurate, fast, simple, highly successful and reproducible. Obturator neurolysis was confirmed as an efficient and cost-effective technique to reduce adductor muscle spasm and related pain and to improve gait and hygienic care in patients with neurological sequelae of stroke, head trauma or any lesion of the motor neurone. PMID- 11900471 TI - Treatment of complex regional pain syndrome type I. AB - Reflex sympathetic dystrophy (RSD), also known as complex regional pain syndrome type I (CRPS I), is a disabling neuropathic pain syndrome. Controversy exists about the effectiveness of therapeutic interventions for the management of RSD/CRPS I. In order to ascertain appropriate therapies we conducted a review of existing randomized controlled trials of therapies for this disabling disease. Eligible trials were identified from the Cochrane, Pubmed, Embase and MEDLINE databases from 1966 through June 2000, from references in retrieved reports and from references in review articles. Twenty-six studies concerning treatment modalities were identified. Eighteen studies were randomized placebo-controlled trials and eight studies were randomized active-controlled trials. Three independent investigators reviewed articles for inclusion criteria using a 15 item checklist. Seventeen of the trials were of high quality according to the 15 item criteria. There was limited evidence for the effectiveness of these interventions because of the heterogeneity of treatment modalities. The search for trials concerning prevention of RSD/CRPS I resulted in two eligible studies. Both were of high quality and dealt with different interventions. There is limited evidence for their preventive effect. PMID- 11900472 TI - Pain behaviour in young immigrants having chronic pain: an exploratory study in primary care. AB - Pain behaviour can hamper rehabilitation. The aim of this study was to explore the phenomenon of pain behaviour in an unselected group of immigrant patients on >6 weeks of sick leave before and after a transcultural treatment programme in primary care. Anxiety about pain and pain behaviour-i.e. > or = 1.5 points on the University of Alabama in Birmingham (UAB) scale with scores of 0-10-were noted before and after treatment. The sex-adjusted odds ratios (OR) for pain behaviour, before and after the treatment, were calculated using logistic regression with 95% confidence intervals (95% CIs). Forty-nine men and 102 women having a mean age of 38 years participated. Their mean sick leave was 46 weeks. All reported psychosocial stress, one-quarter were depressed and social functioning was generally low. The pain was mostly caused by muscular insertion lesions (entesopathies). The frequency of pain behaviour and anxiety about pain declined after treatment (from 68% to 54% and from 76% to 50% respectively). Duration of full-time sick leave and among men also decreasing social functioning were correlated with the UAB score. Those who reported persistent anxiety about pain, or men who were depressed, had higher scores. Only persons on full-time sick leave >1 year had a significant OR for pain behaviour before treatment (OR 3.23; 95% CI 1.17-8.85, adjusted for sex). After treatment, only persons reporting persistent anxiety about pain showed a significant OR for pain behaviour (OR 3.05; 95% CI 1.49-6.23, adjusted for sex). In conclusion, pain behaviour was common in this group of immigrant patients < or = 45 years of age on long-term sick leave. Anxiety about pain and full-time sick leave for more than 1 year significantly predicted pain behaviour. PMID- 11900473 TI - Prevalence of pain in the Spanish population: telephone survey in 5000 homes. AB - Pain has become the most common accompanying symptom in patients seeking medical advice, and it is one of the main issues in public health. In Spain, there are no reliable data about the impact of pain in general population. The aim of the study was to estimate the prevalence of acute and chronic pain in the Spanish general population. An epidemiological observational population-based cross sectional study was carried out by means of a telephone survey. Multistep stratified quota-adjusted sampling was performed with people aged 18-95 years. A computer-assisted questionnaire was administered, covering physical pain symptoms, site, frequency, perceived cause, therapeutic measures and interference with daily life activities. There were 11,980 useful contacts, with 5000 effective interviews (42% of useful sample). Of the interviewees, 29.6% (95% confidence interval, 28.3-30.8%) reported having had pain the day before (women, 37.6%; men, 20.9%) and 43.2% the week before. Most common pain sites were lower extremities (22.7%) and back (cervical and lumbar levels) (21.5%), followed by head (20.5%). Frequency of pain increased with age, reaching 42.6% for people older than 65 years. Among people complaining of pain during the last day or week, duration of symptoms was higher than 3 months in 54% (chronic pain), representing 23.4% of the Spanish general population; most common causes of chronic pain were arthritis, rheumatism and migraine. Regarding treatment, 61.7% of people complaining of pain said they were taking drugs. Source of drug treatment advice was a physician or a nurse in 66.4% of cases and self prescription in 29%. It is concluded that pain, particularly chronic pain, has a high prevalence in the Spanish general population and a significant impact on occupational and social relationships. PMID- 11900474 TI - Direct and indirect costs of managing patients with musculoskeletal pain challenge for health care. AB - Musculoskeletal pain is an outstanding symptom among the patients of primary health care. However, there are few studies of management and costs of musculoskeletal pain at primary health care level. The aim of this study was to describe the diagnostic investigations, management, referral rate and sick leaves related to visits prompted by musculoskeletal pain as well as to assess their costs. A total of 28 general practitioners (GPs) at 25 randomly selected health centres throughout Finland collected the data for this 4 week study, which covered 1 week from each of the four seasons. All visits, except those occurring after hours, were recorded. Altogether 1123 patients visited GPs because of musculoskeletal pain. Laboratory tests were ordered for 12% and imaging investigations for 24%. A total of 16% of the patients suffering from musculoskeletal pain received a prescription for physiotherapy, and analgesics were prescribed to 61% of them. Physicians referred 7% of the pain patients to specialist care. One out of every four patients was prescribed sick leave. The mean cost of the investigations, therapy, referrals, and sick leaves was as high as 530 EUR per visit, with absenteeism from work constituting two-fifths of the total costs. Musculoskeletal pain is not just a frequent complaint but also has extensive economic consequences for society. Investigations and therapy at the primary health care level play a minor role in the costs as compared with specialist care and sick leaves. PMID- 11900475 TI - Somatosensory perception in a remote pain-free area and function of diffuse noxious inhibitory controls (DNIC) in patients suffering from long-term trapezius myalgia. AB - In patients with localized musculoskeletal pain, spread of pain and tenderness outside the primarily painful area and sometimes even generalization of pain have been reported, the latter possibly indicating a dysfunction of endogenous pain modulatory systems. The purpose of the study was to use patients with long-term trapezius myalgia as a model to investigate the possible influence of a localized muscle pain on somatosensory processing in a remote pain-free area and the effect of heterotopic noxious conditioning stimulation (HNCS) on 'diffuse noxious inhibitory controls' (DNIC) related mechanisms. Altered somatosensory processing may indicate subclinical derangement of endogenous modulatory systems. Ten patients with long-term (> or = 1 year) trapezius myalgia and 10 age- and sex matched healthy controls participated. Pressure pain sensitivity, low threshold mechanoreceptive function and thermal sensitivity, including thermal pain, were assessed at the right thigh before, during and following HNCS. Pain was induced in the forearm by the tourniquet test. At rest allodynia to pressure was found at the thigh in conjunction with hypoaesthesia to cold (p<0.03 and p<0.01 respectively), in patients compared with controls. During HNCS, the sensitivity to pressure pain and suprathreshold heat pain decreased in patients and controls alike (p<0.02 and p<0.04 respectively) and returned to baseline following HNCS. In conclusion, in a remote non-painful area allodynia to pressure and hypoaesthesia to cold were found in conjunction with preserved function of DNIC related mechanisms. Whether altered central somatosensory processing at rest may indicate a predisposition for further spread of pain is at present unclear. PMID- 11900476 TI - Somatosensory perception and function of diffuse noxious inhibitory controls (DNIC) in patients suffering from rheumatoid arthritis. AB - The purpose was to investigate the influence of ongoing pain from an inflammatory nociceptive pain with two different disease durations on somatosensory functions and the effect of heterotopic noxious conditioning stimulation (HNCS) on 'diffuse noxious inhibitory controls' (DNIC) related mechanisms. Eleven patients with rheumatoid arthritis of a short duration (<1 year) (RA1), and 10 patients with rheumatoid arthritis of longer duration (>5 years) (RA5) as well as 21 age- and sex-matched healthy controls participated. Pressure pain sensitivity, low threshold mechanoreceptive function and thermal sensitivity, including thermal pain, were assessed over a painful and inflamed joint as well as in a pain-free area, i.e. the right thigh before HNCS (cold-pressor test) and repeated at the thigh only during and following HNCS. In RA1 and RA5 allodynia to pressure was seen over the joint (p<0.02 and p<0.001 respectively) in conjunction with hypoaesthesia to light touch (p<0.02) and hyperaesthesia to innocuous cold (p<0.05) in RA5. At the thigh, allodynia to pressure was found in RA5 (p<0.002). During HNCS, the sensitivity to pressure pain decreased in patients and controls alike (p<0.001). In conclusion, over an inflamed joint allodynia to pressure was found in both RA groups, with additional sensory abnormalities in RA5. In a non painful area, allodynia to pressure was found in RA5, suggesting altered central processing of somatosensory functions in RA5 patients. The response to HNCS was similar in both RA groups and controls, indicating preserved function of DNIC related mechanisms. PMID- 11900477 TI - Effects of collagen IV on neuroblastoma cell matrix-related functions. AB - Integrin-mediated interactions with collagen IV and its domains were examined in a human neuroblastoma cell line (SK-N-SH). By adhesion assays we demonstrated that neuroblastoma cells bound to solid-phase intact collagen IV and synthetic cell-binding peptide HEP-III, derived from the collagenous part of the molecule, but not to the main noncollagenous NC1 domain or to the synthetic cell-binding peptide HEP-I, derived from this domain. Monoclonal antibodies against beta1, alpha3, and alpha(v)beta3 integrins resulted in inhibition of cell binding to collagenous substrates by 95, 30, and 35%, respectively. By flow cytometry and immunoblotting it was shown that culture of SK-N-SH cells on collagen IV resulted in alteration in the expression of major neuroblastoma cell integrins. Binding to collagen IV induced the expression and activation of matrix metalloproteinases A and B (MMP-2, MMP-9), with a concomitant increase at the protein level of tissue specific inhibitors of metalloproteinases (TIMP-1, TIMP-2). Finally, the expression of MMP-2 was significantly up-regulated by anti-alpha3beta1 antibodies, whereas ligation of anti-alpha(v)beta3 antibodies resulted in a modest down-regulation of MMP-2. Our results indicate that the presence of collagen IV modulates the expression of integrins, which are used for binding to this glycoprotein, and MMP-2 secreted by SK-N-SH cells. PMID- 11900478 TI - dlk modulates mitogen-activated protein kinase signaling to allow or prevent differentiation. AB - The EGF-like membrane protein dlk plays a crucial role in the control of cell differentiation. Overexpression of the protein prevents, whereas inhibition of its expression increases, adipocyte differentiation of 3T3-L1 cells in response to Insulin-like Growth Factor I (IGF-1) or insulin. We have investigated whether dlk modulates the signaling pathways known to control this process. We found that the levels of dlk expression modulated signaling through the IGF-1 receptor, causing changes in the activation levels and kinetics of Extracellular-Regulated Kinase/Mitogen-Activated Protein Kinase (ERK/MAPK) that correlated with differentiation outcome. These changes occurred in response to IGF-1 or insulin but not in response to Epidermal Growth Factor. However, the levels of expression of IGF-1 receptor, or the activation of Insulin Receptor Substrate-1 in response to IGF-1, were not affected by the levels of dlk expression. Therefore, dlk appears to modulate ERK/MAPK signaling in response to specific differentiation signals. PMID- 11900479 TI - Visualization of transcription-dependent association of imprinted genes with the nuclear matrix. AB - Genomic imprinting is characterized by allele-specific gene expression as a biological phenomenon. To analyze the participation of the nuclear matrix in the expression of imprinted genes, we first examined the allelic expression state of genes by simultaneously visualizing their primary transcripts and the gene sequences in individual cell nuclei using fluorescence in situ hybridization (FISH). We confirmed that each imprinted gene, SNRPN and UBE3A in human lymphocytes and Igf2 and H19 in mouse embryonic fibroblasts, mainly expressed from one allele, although some nuclei showed biallelic expression. We next visualized the gene sequences on the nuclear matrix by FISH with a tyramide signal amplification technique. Interestingly, we predominantly observed one DNA signal of imprinted genes on the nuclear matrix preparation, closely correlated with their expression patterns. Using patient cells, we confirmed that both the transcription and the binding to the nuclear matrix of the SNRPN gene occurred at the paternal allele. Our results suggest that the nuclear matrix plays an important role in gene expression, including imprinted genes, and that the FISH technique used here allows us to visualize the behaviors of genes at an individual cell level. PMID- 11900480 TI - Polymorphonuclear leukocyte-myocyte interaction: an early event in collar-induced rabbit carotid intimal thickening. AB - The importance of mononuclear leukocyte (MO) adhesion to dysfunctional endothelium and migration to the subendothelial space in the early phases of atherogenesis is well established. Few studies have addressed the relevance of polymorphonuclear leukocytes (PMNs) in the context of evolving lesions, and nothing is known about PMN interaction with vascular smooth muscle cells (SMCs). In this study, we investigated leukocyte/SMC interactions in a model of rabbit carotid injury induced by placement of a silastic collar. This procedure leads to the development of intimal thickening characterized by SMC accumulation preceded by an abundant leukocyte infiltration. By transmission electron microscopy and immunocytochemistry, we demonstrated the occurrence of PMN infiltration starting at 6 h and ceasing within 72 h after collar placement. A previously unknown extensive interaction between medial myocytes and PMNs was detected, referring to emperipolesis, an active phenomenon of cells engulfing other cells in a process other than phagocytosis. PMNs, but not MOs, were internalized by SMCs, which showed ultrastructural features intermediate between the true contractile and the fully synthetic phenotype without exhibiting any sign of injury. Emperipolesis preceded any detectable cell proliferation in the vessel wall and disappeared within 72 h, following the kinetic of PMN infiltration in the vessel wall. In summary, our findings show the occurrence of an active and selective interaction between PMNs and SMCs via emperipolesis during the early phases of intimal thickening after perivascular collaring. However, the overall etiology of the phenomena described in the present study and their pathophysiological significance should be further investigated. PMID- 11900481 TI - Norepinephrine specifically stimulates ribonucleotide reductase subunit R2 gene expression in proliferating brown adipocytes: mediation via a cAMP/PKA pathway involving Src and Erk1/2 kinases. AB - We have examined whether a qualitative switch occurs in the response of the ribonucleotide reductase (RNR) genes to the effect of the physiological cAMP elevating agent norepinephrine (NE) during the development of brown adipocytes. Basal expression of the genes for both RNR subunits, R1 and R2, was high in proliferating cells, but was markedly down-regulated in parallel with adipocyte differentiation. NE stimulation, which promotes DNA synthesis and proliferation of brown preadipocytes, resulted in an increased expression of the R2 gene in proliferating cells (1.6-fold), but was without effect on R1 expression. In contrast, NE stimulation of confluent differentiating brown adipocytes reduced both R1 and R2 expression. The NE stimulation of R2 expression in preadipocytes was mimicked by forskolin and abolished by H89, demonstrating mediation via cAMP and protein kinase A (PKA). Also, inhibitors of Src and of Erk1/2 kinases markedly reduced NE-stimulated R2 expression. We conclude that adrenergic stimulation of brown adipocytes by NE specifically elevates expression of the RNR subunit R2 gene in the proliferative stage of brown adipocyte development, the mediating pathway being a cAMP/PKA cascade further involving Src and the MAP kinase Erk1/2. These results suggest that adrenergic stimulation of brown adipocyte proliferation may act at the level of gene expression of the limiting subunit for RNR activity, R2, and demonstrate a qualitative switch in the response of the R2 gene to cAMP-elevating agents as a consequence of the switch from proliferating to differentiating cell status. PMID- 11900483 TI - Overexpression of BMP-2 modulates morphology, growth, and gene expression in osteoblastic cells. AB - Bone morphogenetic proteins (BMP) play a pivotal role in growth and differentiation of osteoblastic lineage cells. BMPs are potent stimulators of bone formation in various animal models. To understand the mechanism of BMP action in bone cells, we have investigated the effects of overexpression of the BMP-2 gene on proliferation and differentiation of UMR-106 rat osteosarcoma cells. A stable UMR-106 cell line overexpressing the BMP-2 gene was established by transfection of cells using a mammalian expression vector harboring human BMP 2 cDNA followed by G418 selection. After introduction of the BMP-2 gene, UMR-106 cells appeared more spindle-shaped in morphology compared to the predominantly cuboidal appearance of the parental cells. Overexpression of BMP-2 markedly inhibited proliferation as measured by cell counting and [3H]thymidine incorporation assays. Extracellular matrix (ECM) derived from cells overexpressing BMP-2 exhibited a less supportive effect on proliferation of UMR cells than did ECM derived from parental cells. Furthermore, cell-cell communication through gap junctions was reduced more than 50% as determined by nondisruptive fluorescent dye transfer assays. Overexpression of BMP-2 significantly stimulated expression of osteocalcin and alkaline phosphatase genes, indicating its role in osteoblastic differentiation. There was little effect on osteopontin gene expression. PMID- 11900482 TI - Connexins 43 and 26 are differentially increased after rat bladder outlet obstruction. AB - To evaluate the regulation of connexin expression by fluid pressure, we have studied the effects of elevated transmural urine pressure on Connexin43 (Cx43) and Cx26. We chose to focus on these two proteins out of the five connexins (Cx26, 43, 40, 37, and 45) which we found by RT-PCR to be expressed in the rat bladder, since in situ hybridization and immunofluorescence showed that Cx43 is the predominant connexin expressed by smooth muscle cells (SMC), whereas Cx26 is abundantly expressed only in the latter cell type. To evaluate whether these connexins are affected by changes in transmural urine pressure, we used a rat model of bladder outlet obstruction, in which a ligature is placed around the urethra. Under conditions of increased fluid pressure due to urine retention, we observed that the expression of both Cx43 and Cx26 increased at both transcript and protein levels, reaching a maximum 7-9 h after the ligature. Further analysis revealed that these changes were accounted for by a fourfold increase in Cx43 mRNA of SMC but not urothelial cell and by a fivefold increase in Cx26 mRNA of urothelium. Scrape-loading of propidium iodide showed that the latter change was paralleled by a twofold increase in coupling between urothelial cells. The data show that Cx43 and Cx26 are differentially regulated during bladder outlet obstruction and contribute to the response of the bladder wall to increased voiding pressure, possibly to control its elasticity. PMID- 11900484 TI - Immunoreactivity to cell surface syndecans in cytoplasm and nucleus: tubulin dependent rearrangements. AB - Syndecans are transmembrane proteoglycans implicated in the regulation of cell growth and differentiation, by interacting with growth factors. Although syndecans play a major role in regulating cell morphology, little is known about their subcellular distribution and in vivo association with the cytoskeleton. To address this question, we investigated the subcellular distribution and dynamic rearrangement of syndecans-1, -2, and -4, using confocal laser microscopy. Furthermore, we monitored the spatial relation of syndecans to tubulin in both mitotic and interphase cells. Initially, the reactivity to syndecans was confined to the cytoplasm, staining of the cell membranes appearing later. Syndecan-1 also seems to translocate to the nucleus in a time-dependent manner. The mitotic spindle shows unexpectedly more syndecans than that found in interphase cells. After vinblastine treatment, both syndecan-1 and tubulin were recovered as paracrystalline occlusion bodies, and the nuclear reactivity to syndecan-1 disappeared, suggesting tubulin-mediated nuclear transport of this proteoglycan. Plasma membrane staining reappeared in the postmitotic cells. Nuclear translocation predominantly affected syndecan-1, whereas syndecan-2 and -4 remained in cytoplasm and cell membrane. This is the first report on regulated nuclear translocation and the presence of syndecan-1 in the mitotic spindle, where it may stabilize the mitotic machinery. The syndecan-1/tubulin complex may also act as a vehicle for the transport of protein growth factors to the cell nucleus. PMID- 11900485 TI - Hsp70 family member, mot-2/mthsp70/GRP75, binds to the cytoplasmic sequestration domain of the p53 protein. AB - Hsp70 family member mot-2/mthsp70/GRP75/PBP74 was shown to bind to the tumor suppressor protein p53. In this study, by in vivo coimmunoprecipitation of mot-2 with p53 and its deletion mutants, the mot-2 binding site of p53 was mapped to its C-terminal amino acid residues 312-352, a region of p53 that includes its cytoplasmic sequestration domain. These data demonstrate that cytoplasmic sequestration and inactivation of p53 by mot-2 occurs by its binding to the cytoplasmic sequestration domain. Therefore, perturbation of mot-p53 interactions can be employed to abrogate cytoplasmic retention of wild-type p53 in tumors. PMID- 11900486 TI - Intracellular localization of endothelial cell annexins is differentially regulated by oxidative stress. AB - Annexins are calcium-dependent phospholipid binding proteins that are implicated in the regulation of both intracellular and extracellular thrombostatic mechanisms in the vascular endothelium. Tight control of annexin gene expression and targeting of annexin proteins is therefore of importance in maintaining the health of the endothelium. Because annexins are abundant in vascular endothelial cells and could be either dysregulated by or contribute to anomalies in Ca2+ signaling, we investigated annexin gene expression and subcellular localization in human umbilical vein endothelial cells (HUVEC) in a model of chronic oxidative stress. HUVEC were cultured under mild hyperoxic conditions in a custom-built chamber to induce oxidative stress over a period of 12 days. Although annexin expression levels did not change significantly in response to hyperoxic stress, immunofluorescence analysis revealed striking effects on the subcellular localization of certain annexins, including the redistribution of annexins 5 and 6 from the cytosol to the nucleus. In addition, oxidative stress modulated the responses of certain annexins to stimulation with a range of pharmacological and physiological Ca2+-mobilizing agonists, in a manner that suggested that annexin localization is regulated via the complex integration of both Ca2+ and intracellular signaling pathways. These results show that differential regulation of annexin localization by oxidative stress may have a causative role in the cellular pathophysiology of vascular endothelial cell disease. PMID- 11900487 TI - Migration of vascular smooth muscle cells induced by sphingosine 1-phosphate and related lipids: potential role in the angiogenic response. AB - The bioactive lipids sphingosine 1-phosphate (SPP), sphingosylphosphorylcholine, and lysophosphatidic acid play an important role in angiogenesis as a result of their effects on both the migration of endothelial cells (ECs) and the integrity of EC monolayers. Here we show that extremely low concentrations of serum and nanomolar concentrations of these biologically active lipids stimulate migration of human aortic smooth muscle cells (SMCs). However, at dosages most effective in promoting EC migration and in enhancing EC monolayer integrity, serum and SPP potently inhibited SMC migration; SPP also blocked the migration induced by protein growth factors. Treatment of SMCs with SPP induced transient phosphorylation of a 175- to 185-kDa protein corresponding to the PDGF receptor, indicating transactivation of this receptor. SPP and related lipids may play a key role in angiogenesis by coordinating the migration of both endothelial cells and vascular smooth muscle cells in response to the changing gradients of these bioactive lipid messengers. PMID- 11900488 TI - Effects of establishing cell cultures and cell culture conditions on the proliferative life span of human fibroblasts isolated from different tissues and donors of different ages. AB - It is well known that normal human cells placed in a culture environment exhibit a limited proliferative capacity. The extent to which the culture environment influences proliferative life span is not understood. This study evaluated the effects of the standard procedures used to establish and maintain cultures on the proliferative life spans of different types of human fibroblast cells established from fetal and adult skin and lung. The results of this study demonstrate that procedures to establish cell cultures use only one of several subpopulations of cells present in biopsy pieces and that the culture conditions routinely employed by most laboratories can exert significant effects on proliferative life-span determinations. The maximum proliferative life span differed significantly when obtained by growing the cells in two commonly used commercial media. Proliferative life span was inversely related to ambient oxygen tension and directly related to seeding density in all of the lines examined although lines established from adult skin were much more resistant to toxicity. Enzymatic antioxidant defense levels of fetal skin fibroblasts were much lower than those observed in adult skin fibroblasts, but the effects of oxygen on their life spans were similar. Hyperoxia induced larger increases in glutathione concentration in cell lines with low antioxidant enzyme levels. PMID- 11900489 TI - Preferential binding of Grb2 or phosphatidylinositol 3-kinase to the met receptor has opposite effects on HGF-induced myoblast proliferation. AB - Hepatocyte growth factor (HGF) and its receptor, Met, play a crucial role in regulating adult skeletal myoblast proliferation and differentiation. Met signaling is mediated by phosphorylation of two carboxy-terminal tyrosines, which act as docking sites for a number of intracellular mediators. These include Grb2 and p85, which couple the receptor with the Ras and phosphatidylinositol 3-kinase (PI3K) pathways, respectively. In this study, we define the role of these effectors in response to HGF by utilizing Met mutants, designed to obtain preferential coupling of Met to either Grb2 or PI3K or both. We found that relative to the wild-type receptor, enhanced binding to Grb2 further increases the incorporation of bromodeoxyuridine and the expression of Twist, while decreasing that of p27(Kip1) and myogenin. Conversely, preferential coupling with PI3K induced cell-cycle withdrawal and differentiation. Whereas enhanced Grb2 binding increased the phosphorylation of the mitogen-activated protein kinase/extracellular signal-regulated protein kinases (MAPK/ERK) and abrogated that of p38 MAPK, PI3K had the opposite effect. PD098059 reversed the inhibitory effects of Met on cell proliferation and differentiation, while wortmannin had only a very marginal effect. Taken together, these data suggest that coupling of Met with Grb2 is necessary for HGF-mediated inhibition of muscle differentiation. This inhibition occurs only when PI3K signaling downstream of Met is low. Imposing an efficient coupling of PI3K to Met would lead to upregulation of muscle regulatory factors and subsequent cell differentiation. PMID- 11900490 TI - Human dermal fibroblasts escape from the long-term phenocopy of senescence induced by psoralen photoactivation. AB - We have previously shown that following psoralen photoactivation (PUVA treatment) human dermal fibroblasts undergo long-term growth arrest as well as morphological and functional changes reminiscent of cellular senescence [ 1 ]. In the absence of molecular data on what constitutes normal senescence, it has been difficult to decide whether these PUVA-induced changes reflect cellular senescence or rather a mimic thereof. We herein report that PUVA-induced growth arrest, the senescent phenotype with long-term induction of senescence-associated beta-galactosidase, as well as increased expression of matrix metalloprotease-1 are fully reversible at days 100 to 130 post PUVA treatment in four independently tested fibroblast strains. The late returning growth capacity in PUVA-treated fibroblasts is not due to immortalization, as shown by continued lack of telomerase activity, accelerated telomere shortening, and a decrease in overall growth rates in fibroblasts in their regrowing phase post PUVA treatment. Lack of anchorage independent growth additionally suggests that the cells are also not tumorigenically transformed. Collectively, our data suggest that PUVA-induced changes do not fully reflect replicative senescence but rather represent a long term transient phenocopy of senescence. The model reported here is particularly suited to elucidating mechanisms underlying long-term transient growth arrest, the related functional changes, and the release of cells thereof. PMID- 11900491 TI - Enhanced fibroblast contraction of 3D collagen lattices and integrin expression by TGF-beta1 and -beta3: mechanoregulatory growth factors? AB - Generation of contractile forces as fibroblasts attach and migrate through collagenous substrates is a fundamental behavior, yet its regulation and consequences are obscure. Although the transforming growth factor-betas (TGF beta) are similarly important in fibrosis and tissue repair, their role in contraction is controversial. Using a quantitative, 3D collagen culture model we have measured the effects of TGF-beta1 and -beta3 on contractile forces generated by human dermal fibroblasts. Maximal stimulation was between 7.5 and 15 ng/ml of TGF-beta1. Higher doses were inhibitory (30 ng/ml), giving a bell-shaped dose response. The initial rate of force generation was increased sevenfold (15 ng/ml). A similar response pattern was seen with TGF-beta3 alone. However, the addition of both isoforms together stimulated a biphasic increase in force generation, suggesting that there was a distinct temporal cooperativity between the two isforms. This very early onset (10-20 min) of stimulation suggested that TGF-beta might act through cell attachment and integrin function and the effect of TFG-beta on expression of fibronectin (FnR) and vitronectin (VnR) integrin receptors was monitored over the same time scale. TGF-beta1 dramatically up regulated VnR expression, relative to FnR, over time but the optimal time for this was 2-4 h later than that of force stimulation. It is concluded that TGF beta1 and -beta3 behave here primarily as mechanoregulatory growth factors and that stimulation of integrin expression may be a consequence of the altered cell stress. PMID- 11900492 TI - 1,25-(OH)2-vitamin D3 suppresses the bone-related Runx2/Cbfa1 gene promoter. AB - The steroid hormone 1,25-(OH)2-vitamin D3 (VD3) regulates osteoblast differentiation by either activating or repressing transcription of numerous bone phenotypic genes. We addressed whether VD3 also influences osteogenesis by controlling activity of the runt-related transcription factor Runx2/Cbfa1, a key regulator of bone formation in vivo. Our data showed that expression of Runx2 was downregulated by VD3 within 24 h in MC3T3 and ROS 17/2.8, but not in ROS 24.1 cells, which lack a functional vitamin D receptor (VDR). Transient transfection assays showed that the initial 0.6 kb of the bone-related rat and mouse Runx2 promoters both exhibited a 50% reduction of promoter activity in response to VD3 in osteoblastic cells. Furthermore, VD3 inhibited Runx2 transcription in ROS 24.1 cells only upon forced expression of the VDR. Gel mobility shift assays with antibodies and oligonucleotide competition experiments demonstrated that proximal promoter sequences (-92 to -16) contain a functional VD3-responsive element (VDRE) that binds a VDR/retinoid X receptor heterodimer. Mutation of this VDRE completely abolished responsiveness of the Runx2 promoter to VD3 treatment. Together these studies establish that Runx2 expression is regulated by VD3. This VD3-mediated suppression of Runx2 activity provides regulatory coupling between tissue-specific and steroid hormone-dependent control of genes during bone formation. PMID- 11900493 TI - The binding of ORC2 to chromatin from terminally differentiated cells. AB - Nuclei from terminally differentiated Xenopus erythrocytes lack essential components of the prereplication complex, including the origin recognition complex (ORC) proteins XORC1 and XORC2. In Xenopus egg extract, these proteins are able to bind erythrocyte chromatin from permeable nuclei, but not from intact nuclei, even though they are able to cross an intact nuclear envelope. In this report we use both permeable and intact erythrocyte nuclei to investigate the role of cyclin-dependent kinase activity in modulating the binding of XORC2 to chromatin. We find that elevating the level of cyclin A-dependent kinase in egg extract prevents the binding of XORC2 to chromatin from permeable nuclei and that kinase inhibition reverses this effect. We also observe a nuclear transport dependent accumulation of H1 kinase activity within intact nuclei incubated in the extract. However, inhibiting this kinase activity does not facilitate the binding of XORC2 to chromatin, suggesting that other molecules and/or mechanisms exist to prevent association of XORC proteins with replication origins within intact nuclei from terminally differentiated cells. PMID- 11900494 TI - Clinical characteristics and treatment outcome of early Lyme disease in patients with microbiologically confirmed erythema migrans. AB - BACKGROUND: Lyme disease has a wide spectrum of clinical manifestations. Diagnosis is usually based on the clinical and serologic picture rather than on microbiological confirmation. OBJECTIVE: To examine the clinical presentation and treatment outcome of early Lyme disease in patients with microbiologically confirmed erythema migrans. DESIGN: Observational cohort study. SETTING: 31 university-based or clinician-practice sites in 10 endemic states. PARTICIPANTS: 10 936 participants enrolled in a phase III trial of Lyme disease vaccine; 118 participants had erythema migrans in which Borrelia burgdorferi was detected by culture or polymerase chain reaction. MEASUREMENTS: Clinical characteristics and treatment outcome were noted. Skin biopsies of erythema migrans were performed for culture and detection of B. burgdorferi by polymerase chain reaction; serologic responses were determined by Western blot. RESULTS: The 118 patients with microbiologically confirmed erythema migrans presented a median of 3 days after symptom onset. Early erythema migrans commonly had homogeneous or central redness rather than a peripheral erythema with partial central clearing. The most common associated symptoms were low-grade fever, headache, neck stiffness, arthralgia, myalgia, or fatigue. By convalescence, 65% of patients had positive IgM or IgG antibody responses to B. burgdorferi. Most patients responded promptly to antibiotic treatment. CONCLUSIONS: In major endemic areas in the United States, Lyme disease commonly presents as erythema migrans with homogeneous or central redness and nonspecific flu-like symptoms. Clinical outcome is excellent if antibiotic therapy is administered soon after symptom onset. PMID- 11900495 TI - Efficacy of resident training in smoking cessation: a randomized, controlled trial of a program based on application of behavioral theory and practice with standardized patients. AB - BACKGROUND: New educational programs must be developed to improve physicians' skills and effectiveness in counseling patients about smoking cessation. OBJECTIVE: To assess the efficacy of an educational program based on behavioral theory, active learning methods, and practice with standardized patients in helping patients abstain from smoking and changing physicians' counseling practices. DESIGN: Cluster randomized, controlled trial. SETTING: Two general internal medicine clinics in Switzerland. PARTICIPANTS: 35 residents and 251 consecutive smoking patients. INTERVENTION: A training program administered over two half-days, during which physicians learned to provide counseling that matched smokers' motivation to quit and practiced these skills with standardized patients acting as smokers at different stages of change. The control intervention was a didactic session on management of dyslipidemia. MEASUREMENTS: Self-reported abstinence from smoking at 1 year of follow-up, which was validated by exhaled carbon monoxide testing at one clinic; score of overall quality of counseling based on use of 14 counseling strategies; patient willingness to quit; and daily cigarette consumption. RESULTS: At 1 year of follow-up, abstinence from smoking was significantly higher in the intervention group than in the control group (13% vs. 5%; P = 0.005); this corresponded to a cluster-adjusted odds ratio of 2.8 (95% CI, 1.4 to 5.5). Residents who received the study training provided better counseling than did those who received the control training (mean score, 4.0 vs. 2.7; P = 0.002). Smokers' willingness to quit was also higher in the intervention group (94% vs. 80%; P = 0.007). A nonsignificant trend toward lower daily cigarette consumption in the intervention group was observed. CONCLUSION: A training program in smoking cessation administered to physicians that was based on behavioral theory and practice with standardized patients significantly increased the quality of physicians' counseling, smokers' motivation to quit, and rates of abstinence from smoking at 1 year. PMID- 11900496 TI - J-shaped relationship between blood pressure and mortality in hypertensive patients: new insights from a meta-analysis of individual-patient data. AB - BACKGROUND: Population-based longitudinal studies of hypertension have usually shown a continuous and positive relationship between blood pressure and mortality. However, several studies in hypertensive patients receiving treatment have described this relationship as J-shaped, with an increased risk for events in patients with low blood pressure. OBJECTIVE: To assess the evidence for a J shaped relationship between blood pressure and mortality and its relation to treatment. DESIGN: Meta-analysis of individual-patient data. SETTING: Seven randomized clinical trials from the INDANA (INdividual Data ANalysis of Antihypertensive intervention) database. PATIENTS: 40 233 persons with hypertension (mean follow-up, 3.9 years). INTERVENTION: Primarily beta-blockers or thiazide diuretics versus placebo or no treatment. MEASUREMENTS: Diastolic and systolic blood pressure and number of cardiovascular, noncardiovascular, and all cause deaths in yearly periods of follow-up. RESULTS: The analysis included data on 1655 deaths (56% cardiovascular). A J-shaped relationship between diastolic blood pressure and risk for death was observed for total and cardiovascular mortality in treated patients (nadir, 84 and 80 mm Hg, respectively) and untreated patients (nadir, 90 and 85 mm Hg, respectively). For noncardiovascular deaths, the relationship was J-shaped in the treated group (nadir, 84 mm Hg) and negative in the control group. Similar results were observed for systolic blood pressure. The presence of patients with wide pulse pressure did not explain these findings. CONCLUSIONS: The increased risk for events observed in patients with low blood pressure was not related to antihypertensive treatment and was not specific to blood pressure-related events. Poor health conditions leading to low blood pressure and an increased risk for death probably explain the J-shaped curve. PMID- 11900497 TI - Mixed hepatocellular-cholestatic liver injury after pioglitazone therapy. AB - BACKGROUND: Pioglitazone is an oral hypoglycemic agent in the thiazolidinedione class. Only one case of hepatotoxicity related to this agent has previously been reported. OBJECTIVE: To report the clinical course of a patient with hepatitis after therapy with pioglitazone. DESIGN: Case report. SETTING: A community hospital. PATIENT: A 49-year-old diabetic man taking pioglitazone, 30 mg/d. INTERVENTION: Discontinuation of pioglitazone therapy. MEASUREMENTS: Serum aminotransferase and bilirubin levels, standard blood tests for causes of hepatitis and cirrhosis other than drug toxicity, and liver biopsy. RESULTS: After 6 months of pioglitazone therapy, significant hepatic dysfunction developed. Blood tests excluded viral, metabolic, and autoimmune disorders. Liver biopsy showed mixed hepatocellular-cholestatic injury compatible with drug toxicity. After treatment with pioglitazone was discontinued, liver enzyme values returned to normal. CONCLUSION: Patients receiving pioglitazone may develop serious liver injury and should be observed for evidence of hepatitis. PMID- 11900498 TI - Update in women's health. PMID- 11900499 TI - Long-term medical care of testicular cancer survivors. AB - Testicular cancer is the most common solid tumor diagnosed in men 20 to 35 years of age. Because of highly effective treatments that may include surgery, chemotherapy, and radiation therapy, most patients become long-term survivors. Health-related issues that confront testicular cancer survivors include the late medical effects of chemotherapy, the late relapse of disease, the development of second cancers, the effect of the disease and treatment on fertility, and the psychosocial consequences. This case-based discussion focuses on the primary care physician's evaluation and management of a long-term survivor of testicular cancer who was previously treated with surgery and chemotherapy. PMID- 11900500 TI - Deconstructing the placebo effect and finding the meaning response. AB - We provide a new perspective with which to understand what for a half century has been known as the "placebo effect." We argue that, as currently used, the concept includes much that has nothing to do with placebos, confusing the most interesting and important aspects of the phenomenon. We propose a new way to understand those aspects of medical care, plus a broad range of additional human experiences, by focusing on the idea of "meaning," to which people, when they are sick, often respond. We review several of the many areas in medicine in which meaning affects illness or healing and introduce the idea of the "meaning response." We suggest that use of this formulation, rather than the fixation on inert placebos, will probably lead to far greater insight into how treatment works and perhaps to real improvements in human well-being. PMID- 11900501 TI - Recognition and treatment of erythema migrans: are we off target? PMID- 11900503 TI - On being a patient. On growing old. PMID- 11900502 TI - "Did this drug cause my patient's hepatitis?" and related questions. PMID- 11900504 TI - On being a doctor. An examination of conscience. PMID- 11900505 TI - Diagnosing primary HIV infection. PMID- 11900507 TI - Appropriate use of antibiotics: pharyngitis. PMID- 11900508 TI - Regression and progression of valvulopathy associated with fenfluramine and phentermine. PMID- 11900509 TI - Sleep apnea and gastroesophageal reflux disease. PMID- 11900510 TI - A rollover epidemic in North Dakota from 1994 to 1997. PMID- 11900513 TI - Summary for patients. Rashes and symptoms in early Lyme disease. PMID- 11900514 TI - Summary for patients. Effect of a training program for resident physicians in improving success rate in helping patients quit smoking. PMID- 11900515 TI - Summary for patients. Relationship between blood pressure and death among treated hypertensive patients at the high and low ends of blood pressure control. PMID- 11900516 TI - Summary for patients. Liver damage in a person taking the diabetes drug pioglitazone. PMID- 11900517 TI - A view at the millennium: the efficiency of enzymatic catalysis. AB - Binding TS in preference to S and increasing TDeltaS++by freezing out motions in E X S and E X TS have been accepted as the driving forces in enzymatic catalysis; however, the smaller value of DeltaG++ for a one-substrate enzymatic reaction, as compared to its nonenzymatic counterpart, is generally the result of a smaller value of DeltaH++. Ground-state conformers (E X NACs) are formed in enzymatic reactions that structurally resemble E X TS. E X NACs are in thermal equilibrium with all other E X S conformers and are turnstiles through which substrate molecules must pass to arrive at the lowest-energy TS. TS in E X TS may or may not be bound tighter than NAC in E X NAC. PMID- 11900519 TI - Thermodynamics of DNA interactions from single molecule stretching experiments. AB - On the basis of our analysis of detailed measurements of the dependence of the overstretching transition of double-stranded DNA (dsDNA) on temperature, pH, and ionic strength, we have demonstrated that a model of force-induced melting accurately describes the thermodynamics of DNA overstretching. Measurements of this transition allow us to determine the stability of dsDNA and obtain information similar to that obtained in thermal melting studies. This single molecule technique has the advantage that it can be used to measure DNA stability at any temperature. We discuss the use of this technique to study the nucleic acid chaperone activity of the HIV-1 nucleocapsid protein. PMID- 11900518 TI - Micropattern formation in supported lipid membranes. AB - Phospholipid vesicles exhibit a natural tendency to fuse and assemble into a continuous single bilayer membrane on silica and several other substrate materials. The resulting supported membrane maintains many of the physical and biological characteristics of free membranes, including lateral fluidity. Recent advances, building on the supported membrane configuration, have created a wealth of opportunities for the manipulation, control, and analysis of membranes and the reaction environments they provide. The work reviewed in this Account, which can be broadly characterized as the science and technology of membrane patterning, contains three basic components: lateral diffusion control (barriers), membrane deposition techniques (microarrays), and electric field-induced lateral reorganization. Collectively, these preparative and analytical patterned membrane techniques offer a broad experimental platform for the study and utilization of lipid membranes. PMID- 11900520 TI - From mechanistic studies on artemisinin derivatives to new modular antimalarial drugs. AB - In the first part of this account, the antimalarial drug artemisinin is presented, and the current hypotheses on the mechanism of action of this endoperoxide-based drug are reviewed. The alkylating ability of artemisinin and synthetic analogues toward heme related to their antimalarial efficacy are underlined. Some possible ways for discovery of new drugs, especially the design of trioxaquines, new active molecules recently patented that have been prepared by covalent attachment of a trioxane residue having alkylating ability to a quinoline moiety known to easily penetrate within infected erythrocytes, are presented. PMID- 11900521 TI - The propargylic route as a short and versatile entry to optically active monofluorinated compounds. AB - Using selected models and appropriate NMR techniques, it has been demonstrated that dehydroxyfluorination in the propargylic position can be highly regio- and stereoselective. The corresponding propargylic fluorides are very useful intermediates for short preparations of stereodefined unsaturated or polyunsaturated compounds with a single fluorine atom in allylic or propargylic position. This strategy offers good means for the synthesis of chiral, nonracemic monofluorinated analogues of natural products. PMID- 11900522 TI - The crystal structure of catechol oxidase: new insight into the function of type 3 copper proteins. AB - The crystal structure of catechol oxidase reveals new insight into the functional properties of the type-3 copper proteins. This class of proteins includes the closely related and better-known tyrosinase as well as hemocyanin, an oxygen transport protein. All these proteins have a dinuclear copper center, have similar spectroscopic behaviors, and show close evolutionary and functional relationships. Comparison between the 3D structures of catechol oxidase and hemocyanins reveals the structural reasons for the divergence in function. PMID- 11900523 TI - Quantum theory of dissociative chemisorption on metal surfaces. AB - Recent theoretical progress in gas-surface reaction dynamics, a field relevant to heterogeneous catalysis, is described. One of the most fundamental reactions, the dissociative chemisorption of H2 on metal surfaces, can now be treated accurately using quantum mechanics. Density functional theory is used to compute the molecule-surface interaction, and the motion of the hydrogen atoms is simulated using quantum dynamics, modeling all six molecular degrees of freedom. Theory is in good quantitative agreement with molecular beam experiments, offering useful interpretations, and allowing reliable predictions. The success of the approach calls for extensions to larger systems, such as dissociative chemisorption of polyatomic molecules. PMID- 11900524 TI - Mammalian histidine kinases: do they REALLY exist? PMID- 11900525 TI - Flexible regions within the membrane-embedded portions of polytopic membrane proteins. AB - The conventional view of the structure of the membrane-embedded regions of integral membrane proteins is that they are in contact with lipids that interact with the hydrophobic surfaces of the polypeptide, and therefore have intrinsically rigid alpha-helical structures. Here, we briefly review the evidence that in the case of integral membrane proteins with many membrane spans (including membrane transporters and channels), some membrane peptide segments are more or less completely shielded from the lipid bilayer by other membrane polypeptide portions. These portions do not need to have alpha-helical structures and are likely to be much more flexible than typical membrane-spanning helices. The ability of the band 3 anion exchanger to accommodate anionic substrates of different sizes, geometries, and charge distributions suggests the presence of flexible regions in the active center of this protein. These flexible substructures may have important functional roles in membrane proteins, particularly in the mechanisms of membrane transporters and channels. PMID- 11900526 TI - Molecular crowding accelerates fibrillization of alpha-synuclein: could an increase in the cytoplasmic protein concentration induce Parkinson's disease? AB - Parkinson's disease (PD) is one of many neurodegenerative diseases that are characterized by amyloid fibril formation. Alpha-synuclein is a primary component of the fibrillar neuronal inclusions, known as Lewy bodies, that are diagnostic of PD. In addition, the alpha-synuclein gene is linked to familial PD. Fibril formation by alpha-synuclein proceeds via discrete beta-sheet-rich oligomers, or protofibrils, that are consumed as fibrils grow. Both FPD mutations accelerate formation of protofibrils, suggesting that these intermediates, rather than the fibril product, trigger neuronal loss. In idiopathic PD, other factors may be responsible for accelerating protofibril formation by wild-type alpha-synuclein. One possible factor could be molecular crowding in the neuronal cytoplasm. We demonstrate here that crowding using inert polymers significantly reduced the lag time for protofibril formation and the conversion of the protofibril to the fibril, but did not affect the morphology of either species. Physiologically realistic changes in the degree of in vitro crowding have significant kinetic consequences. Thus, nonspecific changes in the total cytoplasmic protein concentration, induced by cell volume changes and/or altered protein degradation, could promote formation of and stabilize the alpha-synuclein protofibril. PMID- 11900527 TI - Homologous (beta/alpha)8-barrel enzymes that catalyze unrelated reactions: orotidine 5'-monophosphate decarboxylase and 3-keto-L-gulonate 6-phosphate decarboxylase. AB - The 3-keto-L-gulonate 6-phosphate decarboxylase (KGPDC) encoded by the ulaD gene in the Escherichia coli genome [Yew, W. S., and Gerlt, J. A. (2002) J. Bacteriol. 184, 302-306] and orotidine 5'-monophosphate decarboxylase (OMPDC) are homologous (derived from a common ancestor) but catalyze different reactions. The metal independent decarboxylation reaction catalyzed by OMPDC avoids the formation of a vinyl anion intermediate; the Mg2+-dependent decarboxylation reaction catalyzed by KGPDC involves the formation of an enediolate anion intermediate. Based on the available structures of OMPDC, a sequence alignment allows the predictions that (1) KGPDC is a dimer of (beta/alpha)8-barrels, with the active sites located at the dimer interface; (2) KGPDC and OMPDC share an aspartate residue at the end of the first beta-strand and an Asp-x-Lys-x-x-Asp motif at the end of the third beta strand with OMPDC; but (3) KGPDC has a Glu instead of a Lys at the end of the second beta-strand. The structure of KGPDC has been determined in the presence of Mg2+ and the substrate analogue L-gulonate 6-phosphate and confirms these predictions. The carboxylate functional groups at the ends of the first, second, and third beta-strands in KGPDC are ligands of the Mg2+; in OMPDC, the homologues of these residues participate in a hydrogen-bonded network that facilitates the decarboxylation reaction. The 3-OH group of the substrate analogue is coordinated to the Mg2+, supporting the hypothesis that the mechanism of the decarboxylation catalyzed by KGPDC involves stabilization of an enediolate anion intermediate. These structural studies establish the existence of the OMPDC "suprafamily," in which members catalyze reactions that occur in different metabolic pathways and share no mechanistic relationship. The existence of this suprafamily demonstrates that divergent evolution can be opportunistic, conscripting active site features of a progenitor to catalyze unrelated functions. Accordingly, sequence or structure homology alone cannot be used to infer the functions of new proteins discovered in genome projects. PMID- 11900528 TI - Epothilone and paclitaxel: unexpected differences in promoting the assembly and stabilization of yeast microtubules. AB - Paclitaxel (Taxol) and the epothilones are antimitotic agents that promote the assembly of mammalian tubulin and stabilization of microtubules. The epothilones competitively inhibit the binding of paclitaxel to mammalian brain tubulin, suggesting that the two types of compounds share a common binding site in tubulin, despite the lack of structural similarities. It is known that paclitaxel does not stabilize microtubules formed in vitro from Saccharomyces cerevisiae tubulin; thus, it would be expected that the epothilones would not affect yeast microtubules. However, we found that epothilone A and B do stimulate the formation of microtubules from purified yeast tubulin. In addition, epothilone B severely dampens the dynamics of yeast microtubules in vitro in a manner similar to the effect of paclitaxel on mammalian microtubules. We used current models describing paclitaxel and epothilone binding to mammalian beta-tubulin to explain why paclitaxel apparently fails to bind to yeast tubulin. We propose that three amino acid substitutions in the N-terminal region and at position 227 in yeast beta-tubulin weaken the interaction of the 3'-benzamido group of paclitaxel with the protein. These results also indicate that mutagenesis of yeast tubulin could help define the sites of interaction with paclitaxel and the epothilones. PMID- 11900529 TI - Structural and biophysical insights into the role of the insert region in Rac1 function. AB - A 13 amino acid insertion that forms a short 3(10) helix between beta-strand 5 and alpha-helix 4 is a distinguishing feature among most members of the Rho family of GTPases, yet the precise role of this region in signal transduction is poorly understood. Previous in vivo functional studies have implicated the insert region of RhoA, Rac1, and Cdc42 to be important for cell transformation, regulation of the actin cytoskeleton, controlling DNA synthesis, and in the activation of downstream targets. In Rac1, our recent biological studies have suggested that the insert is important for SRF activation and the formation of lamellipodia but is dispensable for all other cellular functions of this protein. In the studies reported herein, we have characterized the effect of the insert deletion on Rac1 structure, thermodynamic stability, and the kinetics of nucleotide association. These in vitro studies help clarify biological data and provide further insights as to the role of this 13 amino acid region in modulating Rac1 function. The studies reveal that deletion of the insert has no effect on Rac1 structure and causes only a marginal (approximately 0.8 kcal/mol) decrease in the DeltaG(fold) of Rac1*GDP*Mg2+. The intrinsic rate of nucleotide dissociation of Rac1*Delta(ins) is decreased by about 1.5-fold compared to that of wild type, and a 3-fold increase in the GEF (Vav2)-mediated exchange rate is observed. In addition, deletion of the insert does not change the K(D) for the interaction of Rac1 with GDI, and similar to that previously observed for Cdc42, no inhibition of GDP dissociation is observed for the deletion mutant relative to that for the native protein. Taken together, the structural and biochemical studies reported here are consistent with our biological data reported previously and suggest that the most likely role of the insert region must be to serve as a binding interface for downstream effectors, particularly those important for actin regulation. PMID- 11900530 TI - Direct interaction of the inhibitory gamma-subunit of Rod cGMP phosphodiesterase (PDE6) with the PDE6 GAFa domains. AB - Retinal rod and cone cGMP phosphodiesterases (PDE6 family) function as the effector enzyme in the vertebrate visual transduction cascade. The activity of PDE6 catalytic subunits is controlled by the Pgamma-subunits. In addition to the inhibition of cGMP hydrolysis at the catalytic sites, Pgamma is known to stimulate a noncatalytic binding of cGMP to the regulatory GAFa-GAFb domains of PDE6. The latter role of Pgamma has been attributed to its polycationic region. To elucidate the structural basis for the regulation of cGMP binding to the GAF domains of PDE6, a photoexcitable peptide probe corresponding to the polycationic region of Pgamma, Pgamma-21-45, was specifically cross-linked to rod PDE6alphabeta. The site of Pgamma-21-45 cross-linking was localized to Met138Gly139 within the PDE6alpha GAFa domain using mass spectrometric analysis. Chimeras between PDE5 and cone PDE6alpha', containing GAFa and/or GAFb domains of PDE6alpha' have been generated to probe a potential role of the GAFb domains in binding to Pgamma. Analysis of the inhibition of the PDE5/PDE6alpha' chimeras by Pgamma supported the role of PDE6 GAFa but not GAFb domains in the interaction with Pgamma. Our results suggest that a direct binding of the polycationic region of Pgamma to the GAFa domains of PDE6 may lead to a stabilization of the noncatalytic cGMP-binding sites. PMID- 11900531 TI - Demonstration of 2:2 stoichiometry in the functional SRI-HtrI signaling complex in Halobacterium membranes by gene fusion analysis. AB - A fusion protein in which the C-terminus of Halobacterium salinarum sensory rhodopsin I (SRI) is connected by a flexible linker to the N-terminus of its transducer (HtrI) was constructed and expressed in H. salinarum. The fusion protein mediated attractant responses to orange light and repellent responses to UV/violet light that were comparable to those produced by the wild-type SRI-HtrI complex. Immunoblot analysis of H. salinarum membrane proteins demonstrated intact fusion protein and no detectable proteolytic cleavage products. Rapid oxidative cross-linking of a monocysteine mutant in the HtrI domain confirmed that the fusion protein exists as a homodimer in the membrane. HtrI-free SRI and HtrI-complexed SRI have been shown previously to exhibit large differences in the pH dependence of their photocycle kinetics and in the pK(a) of Asp76 that controls a pH-dependent spectral transition in SRI. These differences were used to assess whether only one or both SRI domains in the fusion protein were complexed properly to the HtrI homodimer. Measurement of the photochemical activity, the photocycle kinetics, and the absorption spectra at various pH values established that both SRI domains are complexed to HtrI in the fusion protein, and therefore the stoichiometry is 2:2. Closer examination of the HtrI effect on SRI revealed that Asp76 titration in HtrI-free SRI fits two pK(a) values, with 98% and 2% of the molecules titrating with pK(a)'s of 7 and 9, respectively. The same two pK(a)'s of Asp76 are evident in HtrI-complexed SRI, but with 13% with pK(a) of 7 and 87% with pK(a) of 9 and a similar bias toward the pK(a) of 9 in the fusion protein. Titration of the fusion protein with Ala substitution at Arg73, a residue in the photoactive site, in the SRI domain indicates that a basic residue at Arg73 is necessary for the lower pK(a) to be observed. A model in which Arg73 plays a role in the HtrI effect on SRI is discussed. PMID- 11900532 TI - Unique ligand-protein interactions in a new truncated hemoglobin from Mycobacterium tuberculosis. AB - A new truncated hemoglobin (HbO) from Mycobacterium tuberculosis has been expressed and purified. Sequence alignment of HbO with other hemoglobins suggests that the proximal F8 residue is histidine and the distal E7 and the B10 positions are occupied by alanine and tyrosine, respectively. The highly conserved residue at the CD1 position, surprisingly, is tyrosine, making HbO the first exception in the hemoglobin family that does not contain phenylalanine at this position. Resonance Raman data suggest that a strong hydrogen bonding network, involving the B10 Tyr and the CD1 Tyr, stabilizes the heme-bound O2 and CO as evidenced by the relatively low frequency of the Fe-O2 stretching mode (559 cm(-1)) and the high frequency of the Fe-CO stretching mode (527 cm(-1)). The presence of this hydrogen bonding network is supported by mutagenesis studies with the B10 tyrosine or the CD1 tyrosine mutated to phenylalanine. Taken together, these data demonstrate a rigid and polar distal pocket in HbO, which is significantly different from that of HbN, the other hemoglobin from M. tuberculosis. The distinct features in the heme active site structures and the temporal expression patterns of HbO and HbN suggest that these two hemoglobins may have very different physiological functions. PMID- 11900533 TI - Mutational analysis of the subunit interface of Vibrio harveyi bacterial luciferase. AB - Bacterial luciferase is a heterodimeric (alphabeta) enzyme which catalyzes a light-producing reaction in Vibrio harveyi. In addition to the alphabeta enzyme, the beta subunit can self-associate to form a stable but inactive homodimer [Sinclair, J. F., Ziegler, M. M., and Baldwin, T. O. (1994) Nat. Struct. Biol. 1, 320-326]. The studies reported here were undertaken to explore the role of the subunit interface in the conformational stability of the enzyme. To this end, we constructed four mutant heterodimers in which residues at the subunit interface were changed in an effort to alter the volume of an apparent solvent accessible channel at the interface or to alter H-bonding groups. Equilibrium unfolding data for the heterodimer have been interpreted in terms of a three-state mechanism [Clark, C. A., Sinclair, J. F., and Baldwin, T. O. (1993) J. Biol. Chem. 268, 10773-10779]. However, we found that unfolding for the wild-type and mutant luciferases is better described by a four-state model. This change in the proposed mechanism of unfolding is based on observation of residual structure in the subunits following dissociation of the heterodimeric intermediate. All of the mutants display modest reductions in activity but, surprisingly, no change in the DeltaG2H2O value for subunit dissociation and no measurable change in the equilibrium dissociation constant relative to that of the wild-type heterodimer. However, the DeltaG1H2O value for the formation of the dimeric intermediate that precedes subunit dissociation is reduced for three of the mutants, indicating that mutations at the interface can alter the stability of a region of the alpha subunit that is distant from the interface. We conclude that the interface region communicates with the distal domains of this subunit, probably through the active center region of the enzyme. PMID- 11900534 TI - Molecular physiology of phosphoryl group transfer from carbamoyl phosphate by a hyperthermophilic enzyme at low temperature. AB - Enzymes from thermophilic organisms often exhibit low activity at reduced temperature. To obtain a better understanding of this sluggishness, we have studied the reaction at 24 degrees C of the carbamate kinase (CK) from the hyperthermophile Pyrococcus furiosus. This enzyme is much slower at low temperature than is the CK from the mesophile Enterococcus faecalis. X-ray structures demonstrated bound ADP (even when no nucleotide was added) with the hyperthermophilic but not with the mesophilic CK. We use centrifugal gel filtration, rate of dialysis and pulse-chase experiments to demonstrate that the pyrococcal enzyme, at 24 degrees C, binds ADP avidly (K(D) = 34 nM), that ADP dissociates from this complex with a t1/2 value of 2.4 s, and that ADP binding is very fast (kappa = 8.4 x 10(6) M(-1) x s(-1)). The high affinity, rather than restrictions to dissociation, explains the isolation of the pyrococcal enzyme as an ADP complex. Carbamoyl phosphate adds quickly to this complex, and ADP cannot dissociate from the resulting ternary complex, being that it is converted very slowly (t1/2 = 10.3 s) to ATP, which dissociates quickly (t1/2 < 2.4 s). The slow conversion is a part of the normal enzyme reaction and limits the rate of the reaction at 24 degrees C. Thus, the sluggishness of the enzyme at low temperature is not due to slow substrate binding or product release but to the very slow rate of isomerization between enzyme-bound substrates and products. Probably the catalysis of the phosphoryl group transfer is less efficient at low temperature, as suggested by structural data showing that Lys131 is improperly positioned to assist the transfer. PMID- 11900535 TI - Catalysis by entropic effects: the action of cytidine deaminase on 5,6 dihydrocytidine. AB - In neutral solution, 5,6-dihydrocytidine undergoes spontaneous deamination (k25 approximately 3.2 x 10(-5) s(-1)) much more rapidly than does cytidine (k25 approximately 3.0 x 10(-10) s(-1)), with a more favorable enthalpy of activation (DeltaDeltaH# = -8.7 kcal/mol) compensated by a less favorable entropy of activation (TDeltaDeltaS# = -1.8 kcal/mol at 25 degrees C). E. coli cytidine deaminase enhances the rate of deamination of 5,6-dihydrocytidine (kcat/k(non) = 4.4 x 10(5)) by enhancing the entropy of activation (DeltaDeltaH# = 0 kcal/mol; TDeltaDeltaS# = +7.6 kcal/mol, at 25 degrees C). Binding of the competitive inhibitor 3,4,5,6-tetrahydrouridine (THU), a stable analogue of 5,6 dihydrocytidine in the transition state for its deamination, is accompanied by a release of enthalpy (DeltaH = -7.1 kcal/mol, TDeltaDeltaS = +2.2 kcal/mol) that approaches the estimated enthalpy of binding of the actual substrate in the transition state for deamination of 5,6-dihydrocytidine (DeltaH = -8.1 kcal/mol, TDeltaDeltaS = +6.0 kcal/mol). Thus, the shortcomings of THU in capturing all of the binding affinity expected of an ideal transition-state analogue reflect a less favorable entropy of association. That difference may arise from the analogue's inability to displace a water molecule from the "leaving group site" at which ammonia is generated in the normal reaction. The effect on binding of removing the 4-OH group from the transition-state analogue THU, to form 3,4,5,6 tetrahydrozebularine (THZ) (DeltaDeltaH = -2.1 kcal/mol, TDeltaDeltaS = -4.4 kcal/mol), is mainly entropic, consistent with the inability of THZ to displace water from the "attacking group site". These results are consistent with earlier indications [Snider, M. J., and Wolfenden, R. (2001) Biochemistry 40, 11364] that site-bound water plays a prominent role in substrate activation and inhibitor binding by cytidine deaminase. PMID- 11900537 TI - 3'-5' exonuclease of Klenow fragment: role of amino acid residues within the single-stranded DNA binding region in exonucleolysis and duplex DNA melting. AB - The mechanism of the 3'-5' exonuclease activity of the Klenow fragment of DNA polymerase I has been investigated with a combination of biochemical and spectroscopic techniques. Site-directed mutagenesis was used to make alanine substitutions of side chains that interact with the DNA substrate on the 5' side of the scissile phosphodiester bond. Kinetic parameters for 3'-5' exonuclease cleavage of single- and double-stranded DNA substrates were determined for each mutant protein in order to probe the role of the selected side chains in the exonuclease reaction. The results indicate that side chains that interact with the penultimate nucleotide (Q419, N420, and Y423) are important for anchoring the DNA substrate at the active site or ensuring proper geometry of the scissile phosphate. In contrast, side chains that interact with the third nucleotide from the DNA terminus (K422 and R455) do not participate directly in exonuclease cleavage of single-stranded DNA. Alanine substitutions of Q419, Y423, and R455 have markedly different effects on the cleavage of single- and double-stranded DNA, causing a much greater loss of activity in the case of a duplex substrate. Time-resolved fluorescence anisotropy decay measurements with a dansyl-labeled primer/template indicate that the Q419A, Y423A, and R455A mutations disrupted the ability of the Klenow fragment to melt duplex DNA and bind the frayed terminus at the exonuclease site. In contrast, the N420A mutation stabilized binding of a duplex terminus to the exonuclease site, suggesting that the N420 side chain facilitates the 3'-5' exonuclease reaction by introducing strain into the bound DNA substrate. Together, these results demonstrate that protein side chains that interact with the second or third nucleotides from the terminus can participate in both the chemical step of the exonuclease reaction, by anchoring the substrate in the active site or by ensuring proper geometry of the scissile phosphate, and in the prechemical steps of double-stranded DNA hydrolysis, by facilitating duplex melting. PMID- 11900536 TI - Noncysteinyl coordination to the [4Fe-4S]2+ cluster of the DNA repair adenine glycosylase MutY introduced via site-directed mutagenesis. Structural characterization of an unusual histidinyl-coordinated cluster. AB - The Escherichia coli DNA repair enzyme MutY plays an important role in the recognition and repair of 7,8-dihydro-8-oxo-2'-deoxyguanosine-2'-deoxyadenosine (OG*A) mismatches in DNA. MutY prevents DNA mutations caused by the misincorporation of A opposite OG by catalyzing the deglycosylation of the aberrant adenine. MutY is representative of a unique subfamily of DNA repair enzymes that also contain a [4Fe-4S]2+ cluster, which has been implicated in substrate recognition. Previously, we have used site-directed mutagenesis to individually replace the cysteine ligands to the [4Fe-4S]2+ cluster of E. coli MutY with serine, histidine, or alanine. These experiments suggested that histidine coordination to the iron-sulfur cluster may be accommodated in MutY at position 199. Purification and enzymatic analysis of C199H and C199S forms indicated that these forms behave nearly identical to the WT enzyme. Furthermore, introduction of the C199H mutation in a truncated form of MutY (C199HT) allowed for crystallization and structural characterization of the modified [4Fe-4S] cluster coordination. The C199HT structure showed that histidine coordinated to the iron cluster although comparison to the structure of the WT truncated enzyme indicated that the occupancy of iron at the modified position had been reduced to 60%. Electron paramagnetic resonance (EPR) spectroscopy on samples of C199HT indicates that a significant percentage (15-30%) of iron clusters were of the [3Fe-4S]1+ form. Oxidation of the C199HT enzyme with ferricyanide increases the amount of the 3Fe cluster by approximately 2-fold. Detailed kinetic analysis on samples containing a mixture of [3Fe-4S]1+ and [4Fe-4S]2+ forms indicated that the reactivity of the [3Fe-4S]1+ C199HT enzyme does not differ significantly from that of the WT truncated enzyme. The relative resistance of the [4Fe-4S]2+ cluster toward oxidation, as well as the retention of activity of the [3Fe-4S]1+ form, may be an important aspect of the role of MutY in repair of DNA damage resulting from oxidative stress. PMID- 11900538 TI - New model for activation of yeast pyruvate decarboxylase by substrate consistent with the alternating sites mechanism: demonstration of the existence of two active forms of the enzyme. AB - Pyruvate decarboxylase from yeast (YPDC, EC 4.1.1.1) exhibits a marked lag phase in the progress curves of product (acetaldehyde) formation. The currently accepted kinetic model for YPDC predicts that, only upon binding of substrate in a regulatory site, a slow activation step converts inactive enzyme into the active form. This allosteric behavior gives rise to sigmoidal steady-state kinetics. The E477Q active site variant of YPDC exhibited hyperbolic initial rate curves at low pH, not consistent with the model. Progress curves of product formation by this variant were S-shaped, consistent with the presence of three interconverting conformations with distinct steady-state rates. Surprisingly, wild-type YPDC at pH < or =5.0 also possessed S-shaped progress curves, with the conformation corresponding to the middle steady state being the most active one. Reexamination of the activation by substrate of wild-type YPDC in the pH range of 4.5-6.5 revealed two characteristic transitions at all pH values. The values of steady-state rates are functions of both pH and substrate concentration, affecting whether the progress curve appears "normal" or S-shaped with an inflection point. The substrate dependence of the apparent rate constants suggested that the first transition corresponded to substrate binding in an active site and a subsequent step responsible for conversion to an asymmetric conformation. Consequently, the second enzyme state may report on "unregulated" enzyme, since the regulatory site does not participate in its generation. This enzyme state utilizes the alternating sites mechanism, resulting in the hyperbolic substrate dependence of initial rate. The second transition corresponds to binding a substrate molecule in the regulatory site and subsequent minor conformational adjustments. The third enzyme state corresponds to the allosterically regulated conformation, previously referred to as activated enzyme. The pH dependence of the Hill coefficient suggests a random binding of pyruvate in a regulatory and an active site of wild-type YPDC. Addition of pyruvamide or acetaldehyde to YPDC results in the appearance of additional conformations of the enzyme. PMID- 11900539 TI - Role of the low-affinity binding site in electron transfer from cytochrome C to cytochrome C peroxidase. AB - The interaction of yeast iso-1-cytochrome c (yCc) with the high- and low-affinity binding sites on cytochrome c peroxidase compound I (CMPI) was studied by stopped flow spectroscopy. When 3 microM reduced yCc(II) was mixed with 0.5 microM CMPI at 10 mM ionic strength, the Trp-191 radical cation was reduced from the high affinity site with an apparent rate constant >3000 s(-1), followed by slow reduction of the oxyferryl heme with a rate constant of only 10 s(-1). In contrast, mixing 3 microM reduced yCc(II) with 0.5 microM preformed CMPI *yCc(III) complex led to reduction of the radical cation with a rate constant of 10 s(-1), followed by reduction of the oxyferryl heme in compound II with the same rate constant. The rate constants for reduction of the radical cation and the oxyferryl heme both increased with increasing concentrations of yCc(II) and remained equal to each other. These results are consistent with a mechanism in which both the Trp-191 radical cation and the oxyferryl heme are reduced by yCc(II) in the high-affinity binding site, and the reaction is rate-limited by product dissociation of yCc(III) from the high-affinity site with apparent rate constant k(d). Binding yCc(II) to the low-affinity site is proposed to increase the rate constant for dissociation of yCc(III) from the high-affinity site in a substrate-assisted product dissociation mechanism. The value of k(d) is <5 s(-1) for the 1:1 complex and >2000 s(-1) for the 2:1 complex at 10 mM ionic strength. The reaction of horse Cc(II) with CMPI was greatly inhibited by binding 1 equiv of yCc(III) to the high-affinity site, providing evidence that reduction of the oxyferryl heme involves electron transfer from the high-affinity binding site rather than the low-affinity site. The effects of CcP surface mutations on the dissociation rate constant indicate that the high-affinity binding site used for the reaction in solution is the same as the one identified in the yCc*CcP crystal structure. PMID- 11900540 TI - Novel insights into the INK4-CDK4/6-Rb pathway: counter action of gankyrin against INK4 proteins regulates the CDK4-mediated phosphorylation of Rb. AB - The newly discovered oncogenic protein gankyrin, which contains six ankyrin repeats, has been reported to be involved in the phosphorylation and degradation of the retinoblastoma gene product, Rb. Using in vitro systems, we have identified a peptide fragment of gankyrin, 176LHLACDEERN185, which is responsible for binding of gankyrin to Rb. We further demonstrated a different mechanism for gankyrin to facilitate the phosphorylation of Rb, by binding with cyclin dependent kinase 4 (CDK4). This binding does not inhibit the Rb-phosphorylating kinase activity of CDK4, but it competes with p16 binding to CDK4 and counteracts the inhibitory function of p16. We then showed that binding of gankyrin to CDK4 and the consequent counter action of p16 function were not affected by the Rb binding peptide 176LHLACDEERN185, indicating that the two mechanisms are independent. They also involve different structural regions of gankyrin. While the Rb-binding motif is located at the fifth ankyrin repeat, truncation mutants of gankyrin, with the first three or four ankyrin repeats remaining, are sufficient for binding to CDK4 and for counteracting the inhibitory function of p16. These results demonstrate the potential importance of gankyrin in cell cycle control and tumorigenesis and suggest an expanded INK4-CDK4/6-Rb pathway. PMID- 11900541 TI - Indirect determination of the rate of iron uptake into the apoprotein of the ribonucleotide reductase of E. coli. AB - The second-order rate constant k(apo) for uptake of FeII by the apoprotein of Ribonucleotide Reductase R2 has been measured by letting that reaction compete with the uptake of FeII by ferrozine (rate constant k(Fz)). The rate of the FeII/ferrozine reaction was studied at high ferrozine concentrations, and an effective first-order rate constant k(Fz) for the disappearance of FeII determined in the presence of bovine serum albumin as a viscogen. Solutions of apoprotein and ferrozine in various ratios were mixed with FeII solutions in a stopped-flow apparatus, and the growth of the 562 nm FeII(ferrozine)3 absorbance monitored. Attempts to fit the data to a variety of kinetic schemes imply that uptake of the second FeII by apo is slower than uptake of the first, suggesting that the rate-determining step in the activation of R2 is a conformational change after the uptake of the first iron. The resulting value of k(apo) is 1.8(1) x 10(6) M(-1) x s(-1). PMID- 11900542 TI - Molecular features of the copper binding sites in the octarepeat domain of the prion protein. AB - Recent evidence suggests that the prion protein (PrP) is a copper binding protein. The N-terminal region of human PrP contains four sequential copies of the highly conserved octarepeat sequence PHGGGWGQ spanning residues 60-91. This region selectively binds Cu2+ in vivo. In a previous study using peptide design, EPR, and CD spectroscopy, we showed that the HGGGW segment within each octarepeat comprises the fundamental Cu2+ binding unit [Aronoff-Spencer et al. (2000) Biochemistry 40, 13760-13771]. Here we present the first atomic resolution view of the copper binding site within an octarepeat. The crystal structure of HGGGW in a complex with Cu2+ reveals equatorial coordination by the histidine imidazole, two deprotonated glycine amides, and a glycine carbonyl, along with an axial water bridging to the Trp indole. Companion S-band EPR, X-band ESEEM, and HYSCORE experiments performed on a library of 15N-labeled peptides indicate that the structure of the copper binding site in HGGGW and PHGGGWGQ in solution is consistent with that of the crystal structure. Moreover, EPR performed on PrP(23 28, 57-91) and an 15N-labeled analogue demonstrates that the identified structure is maintained in the full PrP octarepeat domain. It has been shown that copper stimulates PrP endocytosis. The identified Gly-Cu linkage is unstable below pH approximately 6.5 and thus suggests a pH-dependent molecular mechanism by which PrP detects Cu2+ in the extracellular matrix or releases PrP-bound Cu2+ within the endosome. The structure also reveals an unusual complementary interaction between copper-structured HGGGW units that may facilitate molecular recognition between prion proteins, thereby suggesting a mechanism for transmembrane signaling and perhaps conversion to the pathogenic form. PMID- 11900543 TI - Mapping the active site-ligand interactions of orotidine 5'-monophosphate decarboxylase by crystallography. AB - The crystal structures of orotidine 5'-monophosphate decarboxylases from four different sources have been published recently. However, the detailed mechanism of catalysis of the most proficient enzyme known to date remains elusive. As the ligand-protein interactions at the orotate binding site are crucial to the understanding of this enzyme, we mutated several of the residues surrounding the aromatic part of the substrate, individually and in combination. The ensuing effects on enzyme structure and stability were characterized by X-ray crystallography of inhibitor, product, or substrate complexes and by chemical denaturation with guanidine hydrochloride, respectively. The results are consistent with the residues K42D70K72D75B being charged and forming an 'alternate charge network' around the reactive part of the substrate. In addition to exerting charge-charge repulsion on the orotate carboxylate, Asp70 also makes a crucial contribution to enzyme stability. Consequently, orotidine 5' monophosphate decarboxylases seem to require the presence of a negative charge at this position for catalysis as well as for correct and stable folding. PMID- 11900544 TI - Formation in vitro of hybrid dimers of H463F and Y74F mutant Escherichia coli tryptophan indole-lyase rescues activity with L-tryptophan. AB - Y74F and H463F mutant forms of Escherichia coli tryptophan indole-lyase (Trpase) have been prepared. These mutant proteins have very low activity with L-Trp as substrate (kcat and kcat/Km values less than 0.1% of wild-type Trpase). In contrast, these mutant enzymes exhibit much higher activity with S-(o nitrophenyl)-L-cysteine and S-ethyl-L-cysteine (kcat/Km values about 1-50% of wild-type Trpase). Thus, Tyr-74 and His-463 are important for the substrate specificity of Trpase for L-Trp. H463F Trpase is not inhibited by a potent inhibitor of wild-type Trpase, oxindolyl-L-alanine, and does not exhibit the pK(a) of 6.0 seen in previous pH dependence studies [Kiick, D. M., and Phillips, R. S. (1988) Biochemistry 27, 7333]. These results suggest that His-463 may be the catalytic base with a pK(a) of 6.0 and Tyr-74 may be a general acid catalyst for the elimination step, as we found previously with tyrosine phenol-lyase [Chen, H., Demidkina, T. V., and Phillips, R. S. (1995) Biochemistry 34, 12776]. H463F Trpase reacts with L-Trp and S-ethyl-L-cysteine in rapid-scanning stopped flow experiments to form equilibrating mixtures of external aldimine and quinonoid intermediates, similar to those observed with wild-type Trpase. In contrast to the results with wild-type Trpase, the addition of benzimidazole to reactions of H463F Trpase with L-Trp does not result in the formation of an aminoacrylate intermediate. However, addition of benzimidazole with S-ethyl-L cysteine results in the formation of an aminoacrylate intermediate, with lambda(max) at 345 nm, as was seen previously with wild-type Trpase [Phillips, R. S. (1991) Biochemistry 30, 5927]. This suggests that His-463 plays a specific role in the elimination step of the reaction of L-Trp. Refolding of equimolar mixtures of H463F and Y74F Trpase after unfolding in 4 M guanidine hydrochloride results in a dramatic increase in activity with L-Trp, indicating the formation of a hybrid H463F/Y74F dimer with one normal active site. PMID- 11900545 TI - Kinetic mechanism of adenine phosphoribosyltransferase from Leishmania donovani. AB - Adenine phosphoribosyltransferase (APRT, EC 2.4.2.7) catalyzes the reversible phosphoribosylation of adenine from alpha-D-5-phosphoribosyl-1-pyrophosphate (PRPP) to form AMP and PP(i). Three-dimensional structures of the dimeric APRT enzyme from Leishmania donovani (LdAPRT) bear many similarities to other members of the type 1 phosphoribosyltransferase family but do not reveal the structural basis for catalysis (Phillips, C. L., Ullman, B., Brennan, R. G., and Hill, C. P. (1999) EMBO J. 18, 3533-3545). To address this issue, a steady state and transient kinetic analysis of the enzyme was performed in order to determine the catalytic mechanism. Initial velocity and product inhibition studies indicated that LdAPRT follows an ordered sequential mechanism in which PRPP is the first substrate to bind and AMP is the last product to leave. This mechanistic model was substantiated by equilibrium isotope exchange and fluorescence binding studies, which provided dissociation constants for the LdAPRT-PRPP and LdAPRT-AMP binary complexes. Pre-steady-state kinetic analysis of the forward reaction revealed a burst in product formation indicating that phosphoribosyl transfer proceeds rapidly relative to some rate-limiting product release event. Transient fluorescence competition experiments enabled measurement of rates of binary complex dissociation that implicated AMP release as rate-limiting for the forward reaction. Kinetics of product ternary complex formation were evaluated using the fluorophore formycin AMP and established rate constants for pyrophosphate binding to the LdAPRT-formycin AMP complex. Taken together, these data enabled the complete formulation of an ordered bi-bi kinetic mechanism for LdAPRT in which all of the rate constants were either measured or calculated. PMID- 11900546 TI - Design and characterization of an improved protein tyrosine phosphatase substrate trapping mutant. AB - Although members of the protein tyrosine phosphatase (PTPase) family share a common mechanism of action (hydrolysis of phosphotyrosine), the cellular processes in which they are involved can be both highly specialized and fundamentally important. Identification of cellular PTPase substrates will help elucidate the biological functions of individual PTPases. Two types of substrate trapping mutants are being used to isolate PTPase substrates. In the first, the active site Cys residue is replaced by a Ser (e.g., PTP1B/C215S) while in the second, the general acid Asp residue is substituted by an Ala (e.g., PTP1B/D181A). Unfortunately, only a limited number of PTPase substrates have been identified with these two mutants, which are usually relatively abundant cellular proteins. Based on mechanistic considerations, we seek to create novel PTPase mutants with improved substrate-trapping properties. Kinetic and thermodynamic characterization of the newly designed PTP1B mutants indicates that PTP1B/D181A/Q262A displays lower catalytic activity than that of D181A. In addition, D181A/Q262A also possesses 6- and 28-fold higher substrate-binding affinity than those of D181A and C215S, respectively. In vivo substrate-trapping experiments indicate that D181A/Q262A exhibits much higher affinity than both D181A and C215S for a bona fide PTP1B substrate, the epidermal growth factor receptor. Moreover, D181A/Q262A can also identify novel, less abundant substrates, that are missed by D181A. Thus, this newly developed and improved substrate-trapping mutant can serve as a powerful affinity reagent to isolate and purify both high- and low-abundant protein substrates. Given that both Asp181 and Gln262 are invariant among the PTPase family, it is predicted that this improved substrate-trapping mutant would be applicable to all members of PTPases for substrate identification. PMID- 11900548 TI - Kinetics and thermodynamics of mandelate racemase catalysis. AB - Mandelate racemase (EC 5.1.2.2) from Pseudomonas putida catalyzes the interconversion of the two enantiomers of mandelic acid with remarkable proficiency, producing a rate enhancement exceeding 15 orders of magnitude. The rates of the forward and reverse reactions catalyzed by the wild-type enzyme and by a sluggish mutant (N197A) have been studied in the absence and presence of several viscosogenic agents. A partial dependence on relative solvent viscosity was observed for values of kcat and kcat/Km for the wild-type enzyme in sucrose containing solutions. The value of kcat for the sluggish mutant was unaffected by varying solvent viscosity. However, sucrose did have a slight activating effect on mutant enzyme efficiency. In the presence of the polymeric viscosogens poly(ethylene glycol) and Ficoll, no effect on kcat or kcat/Km for the wild-type enzyme was observed. These results are consistent with both substrate binding and product dissociation being partially rate-determining in both directions. The viscosity variation method was used to estimate the rate constants comprising the steady-state expressions for kcat and kcat/Km. The rate constant for the conversion of bound (R)-mandelate to bound (S)-mandelate (k2) was found to be 889 +/- 40 s(-1) compared with a value of 654 +/- 58 s(-1) for kcat in the same direction. From the temperature dependence of Km (shown to equal K(S)), k2, and the rate constant for the uncatalyzed reaction [Bearne, S. L., and Wolfenden, R. (1997) Biochemistry 36, 1646-1656], we estimated the enthalpic and entropic changes associated with substrate binding (DeltaH = -8.9 +/- 0.8 kcal/mol, TDeltaS = -4.8 +/- 0.8 kcal/mol), the activation barrier for conversion of bound substrate to bound product (DeltaH# = +15.4 +/- 0.4 kcal/mol, TDeltaS# = +2.0 +/- 0.1 kcal/mol), and transition state stabilization (DeltaH(tx) = -22.9 +/- 0.8 kcal/mol, TDeltaS(tx) = +1.8 +/- 0.8 kcal/mol) during mandelate racemase catalyzed racemization of (R)-mandelate at 25 degrees C. Although the high proficiency of mandelate racemase is achieved principally by enthalpic reduction, there is also a favorable and significant entropic contribution. PMID- 11900547 TI - The intestinal fatty acid binding protein: the role of turns in fast and slow folding processes. AB - The intestinal fatty acid binding protein is one of a family of proteins that are composed of two beta-sheets surrounding a large interior cavity into which the ligand binds. Glycine residues occur in many of the turns between adjacent antiparallel beta-strands. In previous work, the effect of replacing these glycine residues with valine has been examined with stopped flow instrumentation using intrinsic tryptophan fluorescence spectroscopy [Kim and Frieden (1998) Protein Sci. 7, 1821-1828]. To resolve the burst phase missing in the stopped flow measurements, these valine mutants have been reexamined with sub-millisecond continuous flow instrumentation. Some of the glycine residues have also been replaced with proline, and the folding reactions of these proline mutants have been compared with those of their valine counterparts. In all cases, the stability of the protein is decreased, but some turns appear to be more critical for final structure stabilization than others. Surprisingly, the rate constants observed for all the mutants measured by sub-millisecond continuous flow methods are quite similar (1400-3000 s(-1)), and in all the mutants, there is a shift in the fluorescence emission maximum from that of the unfolded protein to lower wavelengths, suggesting some collapse of the unfolded state within 200 micros. In contrast to the rate constants observed for the initial folding events measured by the sub-millisecond continuous flow method, the rate constants for the slower phase observed in the stopped flow instrument vary widely for the different mutants. The latter step appears to be related to side chain stabilization rather than secondary structure formation. It is also shown that the ligand binds tightly only to the native protein and not to any intermediate forms. PMID- 11900549 TI - Pentavalent ions dependency is a conserved property of adenosine kinase from diverse sources: identification of a novel motif implicated in phosphate and magnesium ion binding and substrate inhibition. AB - The catalytic activity of adenosine kinase (AK) from mammalian sources has previously been shown to exhibit a marked dependency upon the presence of pentavalent ions (PVI), such as phosphate (PO4), arsenate, or vanadate. We now show that the activity of AK from diverse sources, including plant, yeast, and protist species, is also markedly enhanced in the presence of PVI. In all cases, PO4 or other PVI exerted their effects primarily by decreasing the Km for adenosine and alleviating the inhibition caused by high concentrations of substrates. These results provide evidence that PVI dependency is a conserved property of AK and perhaps of the PfkB family of carbohydrate kinases which includes AK. On the basis of sequence alignments, we have identified a conserved motif NXXE within the PfkB family. The N and E of this motif make close contacts with Mg2+ and PO4 ions in the crystal structures of AK and bacterial ribokinase (another PfkB member which shows PVI dependency), implicating these residues in their binding. Site-directed mutagenesis of these residues in Chinese hamster AK have resulted in active proteins with greatly altered phosphate stimulation and substrate inhibition characteristics. The N239Q mutation leads to the formation of an active protein whose activity was not stimulated by PO4 or inhibited by high concentrations of adenosine or ATP. The activity of the E242D mutant protein was also not significantly altered in the presence of phosphate. Although PO4 had no effect on the KmAdenosine for this mutant, the KmATP, K(i)Adenosine, and K(i)ATP were significantly decreased. In contrast to these mutations, N239L or E242L mutant proteins showed greatly decreased activity with an altered Mg2+ requirement. These observations support the view that N239 and E242 play an important role in the binding of PO4 and Mg2+ ions required for the catalytic activity of adenosine kinase. PMID- 11900550 TI - Certain metal ions are inhibitors of cytochrome b6f complex 'Rieske' iron-sulfur protein domain movements. AB - Many current models of the Q cycle for the cytochrome (cyt) b6f and the cyt bc1 complexes incorporate 'Rieske' iron-sulfur protein (ISP) domain movements to gate electron transfer and to ensure high yields of proton shuttling. It was previously proposed that copper ions, which bind at a site distant from the quinol oxidase (Q(o)) site, inhibit plastoquinol (PQH2) binding by restraining the hydrophilic head domain of the ISP [Rao B. K., S., Tyryshkin, A. M., Roberts, A. G., Bowman, M. K., and Kramer, D. M. (1999) Biochemistry 38, 3285-3296]. The present work presents evidence that this is indeed the case for both copper ions and Zn2+, which appear to inhibit by similar mechanisms. Electron paramagnetic resonance (EPR) spectra show that Cu2+ and Zn2+ binding to the cyt b6f complex displaces the Q(o) site inhibitor 2,5-dibromo-3-methyl-6-isopropylbenzoquinone (DBMIB). At high concentrations, both DBMIB and Cu2+ or Zn2+ can bind simultaneously, altering the Rieske 2Fe2S cluster and Cu2+ EPR spectra, suggesting perturbations in their respective binding sites. Both Zn2+ and Cu1+ altered the orientations of the Rieske 2Fe2S cluster with respect to the membrane plane, but had no effect on that of the cyt b6 hemes. Cu2+ was found to change the orientation of the cyt f heme plane, consistent with binding on the cyt f protein. Within conservative constraints, the data suggest that the ISP is shifted into a position intermediate between the ISP(C) position, when the Q(o) site is unoccupied, and the ISP(B) position, when the Q(o) site is occupied by inhibitors such as DBMIB or stigmatellin. These results support the role of ISP domain movements in Q(o) site catalysis. PMID- 11900551 TI - Role of aromatic side chains in the binding of volatile general anesthetics to a four-alpha-helix bundle. AB - Currently, the mechanism by which anesthesia occurs is thought to involve the direct binding of inhaled anesthetics to ligand-gated ion channels. This hypothesis is being studied using four-alpha-helix bundles as model systems for the transmembrane domains of the natural "receptor" proteins. This study concerns the role in anesthetic binding played by aromatic side chains in the binding cavity of a four-alpha-helix bundle designed to assume a Rop-like fold. Specifically, the effect of the substitution W15Y on bundle structure, stability, and anesthetic binding energetics was investigated. No appreciable effect of substituting W for Y on the secondary structure or the thermodynamic stability of the four-alpha-helix bundle was identified. However, the substitution W15Y resulted in about 6- and 3-fold decreases in halothane and chloroform binding affinities, respectively. This effect may reflect weaker dipole-aromatic quadrupole interactions between the aromatic side chain and the anesthetic in the tyrosine-containing species, which possesses the smaller aromatic ring system. For these anesthetic binding proteins, this class of interaction occurs when the permanent nonspherical distribution of electrons in the aromatic ring systems interact with the weakly acidic CH group of the anesthetics. PMID- 11900552 TI - Indirect readout of the trp-repressor-operator complex by B-DNA's backbone conformation transitions. AB - Although the trp-repressor-operator complex is one of the best studied transcriptional controlling systems, some questions regarding the specific recognition of the operator by the repressor remain. We performed a 2.35 ns long molecular dynamics simulation to clarify the influence of the two B-DNA backbone conformational substates B(I) and B(II) on complexation. The trp-repressor operator is an ideal biological system for this study because experimental results have already figured out that the interaction between the internucleotide phosphates and the protein is essential for the formation of the high affinity complex. Our simulation supports these results, but more important it shows a strong correlation between the B(I)/B(II) phosphate substate and the number of interactions with this phosphate. In particular the B(I) <==> B(II) transitions occur synchronous to hydrogen bond breaking or formation. To the best of our knowledge, this was observed for the first time. Thus, we conclude that the sequence specific B(I)/B(II) behavior contributes via indirect readout to sequence specific recognition. These results have implication for the design of transcription-controlling drugs in view of the recently published influence of minor groove binders on the B(I)/B(II) pattern. The simulation also agrees with crystallographically observed hydration sites. This is consistent with experimental results and indicates the correctness of the model used. PMID- 11900553 TI - Role of the oxyanion binding site and subsites S1 and S2 in the catalysis of oligopeptidase B, a novel target for antimicrobial chemotherapy. AB - Oligopeptidase B is a member of a novel serine peptidase family, found in Gram negative bacteria and trypanosomes. The enzyme is involved in host cell invasion, and thus, it is an important target for drug design. Oligopeptidase B is specific for substrates with a pair of basic residues at positions P1 and P2. The sensitivity of substrates to high ionic strength suggests that the arginines interact with the carboxylate ions of the enzyme. On the basis of a three dimensional model, two carboxyl dyads (Asp460 and Asp462 and Glu576 and Glu578) can be assigned as binding sites for arginines P1 and P2, respectively. The dyads are involved in several events: (i) substrate binding, (ii) substrate inhibition at high substrate concentrations (different inhibitory mechanisms were demonstrated with substrates bearing one and two arginine residues), (iii) enzyme activation at millimolar CaCl2 concentrations with substrates having one arginine, and (iv) interaction of Ca2+ with the dyads which simplified the complex pH dependence curves. Titration with a product-like inhibitor revealed the pK(a) of the carboxyl group that perturbed the pH-kcat/Km profiles. The OH group of Tyr452 is part of the oxyanion binding site, which stabilizes the transition state of the reaction. Its role studied with the Tyr452Phe variant indicates that (i) the catalytic contribution of the OH group depends on the substrate and (ii) the catalysis is, unusually, an entropy-driven process at physiological temperature. The NH group of the scissile peptide bond accounts for the deviation of the reaction from the Eyring plot above 25 degrees C, and for abolishing potential nonproductive binding. PMID- 11900554 TI - Interaction of fibrin(ogen) with the endothelial cell receptor VE-cadherin: mapping of the receptor-binding site in the NH2-terminal portions of the fibrin beta chains. AB - Interaction of fibrin with endothelial cells stimulates capillary tube formation thus promoting angiogenesis. This interaction occurs via endothelial cell receptor VE-cadherin and fibrin beta chain 15-42 regions [Bach, T. L., et al. (1998) J. Biol. Chem. 272, 30719-30728]. To clarify the mechanism of this interaction, we expressed in Escherichia coli a number of recombinant fibrin(ogen) fragments containing the beta15-42 region or the VE-cad(1-2) and VE cad(1-4) fragments encompassing two and four extracellular NH2-terminal domains of VE-cadherin, respectively, and tested interaction between them by surface plasmon resonance and ELISA. Neither the recombinant Bbeta1-57 or Bbeta1-64 fragments, nor beta15-57 or beta15-64 prepared from the latter fragments by thrombin treatment to remove fibrinopeptides B, bound the recombinant VE-cadherin fragments. At the same time, a dimeric recombinant thrombin-treated (beta15-66)2 fragment, which had been disulfide-linked via Cys65 to mimic the dimeric arrangement of the beta chains in fibrin, bound VE-cad(1-4) well, but not VE cad(1-2); no binding was observed with the untreated (Bbeta1-66)2 dimer. We next mutated several residues in the dimer, His16, Arg17, Pro18, and Asp20, and tested the interaction of the thrombin-treated mutants with VE-cad(1-4) by ligand blotting and surface plasmon resonance. No binding was observed with the H16A and R17Q single mutants and the H16P, P18V double mutant while the P18A and D20N single mutants bound VE-cad(1-4) with the same affinity as the thrombin-treated wild-type dimer. These results indicate that the VE-cadherin binding site in fibrin includes NH2-terminal regions of both fibrin beta-chains, that His16 and Arg17 are critical for the binding, and that the third and/or fourth extracellular domains of VE-cadherin are required for the binding to occur. PMID- 11900555 TI - Electron transfer from the water oxidizing complex at cryogenic temperatures: the S1 to S2 step. AB - We report the detection of a "split" electron paramagnetic resonance (EPR) signal during illumination of dark-adapted (S(1) state) oxygen-evolving photosystem II (PSII) membranes at <20 K. The characteristics of this signal indicate that it arises from an interaction between an organic radical and the Mn cluster of PSII. The broad radical signal decays in the dark following illumination either by back reaction with Qa*- or by forward electron transfer from the Mn cluster. The forward electron transfer (either from illumination at 11 K followed by incubation in the dark at 77 K or by illumination at 77 K) results in the formation of a multiline signal similar to, but distinct from, other well characterized multiline forms found in the S0 and S2 states. The relative yield of the "S1 split signal", which we provisionally assign to S1X*, where X could be YZ* or Car*+, and that of the 77 K multiline signal indicate a relationship between the two states. An approximate quantitation of the yield of these signals indicates that up to 40-50% of PSII centers can form the S1 split signal. Ethanol addition removes the ability to observe the S1 split signal, but the multiline signal is still formed at 77 K. The multiline forms with <700 nm light and is not affected by near-infrared (IR) light, showing that we are detecting electron transfer in centers not responsive to IR illumination. The results provide important new information about the mechanism of electron abstraction from the water oxidizing complex (WOC). PMID- 11900556 TI - Dynamics of energy transfer from lycopene to bacteriochlorophyll in genetically modified LH2 complexes of Rhodobacter sphaeroides. AB - LH2 complexes from Rb. sphaeroides were modified genetically so that lycopene, with 11 saturated double bonds, replaced the native carotenoids which contain 10 saturated double bonds. Tuning the S1 level of the carotenoid in LH2 in this way affected the dynamics of energy transfer within LH2, which were investigated using both steady-state and time-resolved techniques. The S1 energy of lycopene in n-hexane was determined to be approximately 12 500 +/- 150 cm(-1), by direct measurement of the S1-S2 transient absorption spectrum using a femtosecond IR probing technique, thus placing an upper limit on the S1 energy of lycopene in the LH2 complex. Fluorescence emission and excitation spectra demonstrated that energy can be transferred from lycopene to the bacteriochlorophyll molecules within this LH2 complex. The energy-transfer dynamics within the mutant complex were compared to wild-type LH2 from Rb. sphaeroides containing the carotenoid spheroidene and from Rs. molischianum, in which lycopene is the native carotenoid. The results show that the overall efficiency for Crt --> B850 energy transfer is approximately 80% in lyco-LH2 and approximately 95% in WT-LH2 of Rb. sphaeroides. The difference in overall Crt --> BChl transfer efficiency of lyco LH2 and WT-LH2 mainly relates to the low efficiency of the Crt S(1) --> BChl pathway for complexes containing lycopene, which was 20% in lyco-LH2. These results show that in an LH2 complex where the Crt S1 energy is sufficiently high to provide efficient spectral overlap with both B800 and B850 Q(y) states, energy transfer via the Crt S1 state occurs to both pigments. However, the introduction of lycopene into the Rb. sphaeroides LH2 complex lowers the S1 level of the carotenoid sufficiently to prevent efficient transfer of energy to the B800 Q(y) state, leaving only the Crt S1 --> B850 channel, strongly suggesting that Crt S1 -> BChl energy transfer is controlled by the relative Crt S1 and BChl Q(y) energies. PMID- 11900557 TI - Stress-induced aggregation profiles of GST-alpha-synuclein fusion proteins: role of the C-terminal acidic tail of alpha-synuclein in protein thermosolubility and stability. AB - Alpha-synuclein is a well-known heat-resistant protein that does not aggregate upon heat treatment, whereas glutathione S-transferase (GST) is a heat-labile protein that easily precipitates as a result of thermal stress. This paper reports the role of the C-terminal acidic tail of alpha-synuclein in protein thermosolubility and stability. The region of alpha-synuclein that is responsible for the heat resistance was initially investigated using a series of deletion mutants, and the C-terminal acidic tail (residues 96-140) was found to be crucial for the thermosolubility of alpha-synuclein. The thermal behavior of the GST alpha-synuclein fusion protein was next investigated, and the fusion protein was seen to be extremely heat-resistant. Using a series of GST-synuclein deletion mutants, the C-terminal acidic tail of alpha-synuclein was shown to play a critical role in conferring the heat resistance of the fusion proteins. Furthermore, the acidic tail appeared to protect the fusion protein from pH- and metal-induced protein aggregation, suggesting that the acidic tail can increase the virtual stability of the protein by protecting it from the aggregation induced by environmental stresses. Interestingly, the acidic tail also appeared to protect the GST enzyme from the thermal inactivation to a considerable extent. However, CD analysis of the heat-induced secondary structural changes of the GST alpha-synuclein fusion protein revealed that the fusion protein is irreversibly denatured by heat treatment with a slightly lowered melting temperature (Tm). Thus, the results demonstrate that introducing an acidic tail into GST promotes the thermosolubility and virtual stability of the fusion protein, although it might be unfavorable for its intrinsic stability. PMID- 11900558 TI - Substrate specificity engineering of beta-mannosidase and beta-glucosidase from Pyrococcus by exchange of unique active site residues. AB - A beta-mannosidase gene (PH0501) was identified in the Pyrococcus horikoshii genome and cloned and expressed in E. coli. The purified enzyme (BglB) was most specific for the hydrolysis of p-nitrophenyl-beta-D-mannopyranoside (pNP-Man) (Km: 0.44 mM) with a low turnover rate (kcat: 4.3 s(-1)). The beta-mannosidase has been classified as a member of family 1 of glycoside hydrolases. Sequence alignments and homology modeling showed an apparent conservation of its active site region with, remarkably, two unique active site residues, Gln77 and Asp206. These residues are an arginine and asparagine residue in all other known family 1 enzymes, which interact with the catalytic nucleophile and equatorial C2-hydroxyl group of substrates, respectively. The unique residues of P. horikoshii BglB were introduced in the highly active beta-glucosidase CelB of Pyrococcus furiosus and vice versa, yielding two single and one double mutant for each enzyme. In CelB, both substitutions R77Q and N206D increased the specificity for mannosides and reduced hydrolysis rates 10-fold. In contrast, BglB D206N showed 10-fold increased hydrolysis rates and 35-fold increased affinity for the hydrolysis of glucosides. In combination with inhibitor studies, it was concluded that the substituted residues participate in the ground-state binding of substrates with an equatorial C2-hydroxyl group, but contribute most to transition-state stabilization. The unique activity profile of BglB seems to be caused by an altered interaction between the enzyme and C2-hydroxyl of the substrate and a specifically increased affinity for mannose that results from Asp206. PMID- 11900559 TI - Ultrafast electron transfer in the complex between fluorescein and a cognate engineered lipocalin protein, a so-called anticalin. AB - Anticalins are a novel class of engineered ligand-binding proteins with tailored specificities derived from the lipocalin scaffold. The anticalin FluA complexes fluorescein as ligand with high affinity, and it effects almost complete quenching of its steady-state fluorescence. To study the underlying mechanism, we have applied femtosecond absorption spectroscopy, which revealed excited-state electron transfer within the FluA*Fl complex to be responsible for the strong fluorescence quenching. On the basis of a comparison of redox potentials, either tryptophan or tyrosine may serve as electron donor to the bound fluorescein group in its excited singlet state, thus forming the fluorescein trianion radical within 400 fs. The almost monoexponential rate points to a single, well-defined binding site, and its temperature independence suggests an (almost) activationless process. Applying conventional electron transfer theory to the ultrafast forward and slower back-rates, the resulting electronic interaction is rather large, with approximately 140 cm(-1) for tyrosine, which would be consistent with a coplanar arrangement of both aromatic moieties within van der Waals distance. The weak residual steady-state fluorescence originates from a small (approximately 10%) component with a time constant in the 40-60 ps range. These results demonstrate the power of time-resolved absorption spectroscopy as a diagnostic tool for the elucidation of a fluorescence quenching mechanism and the temporal profiles of the processes involved. The high structural and dynamic definition of the complexation site suggests the anticalin FluA to be a promising model in order to tailor and probe electronic interactions and energetics in proteins. PMID- 11900560 TI - Cholesterol modulates interaction between an amphipathic class A peptide, Ac-18A NH2, and phosphatidylcholine bilayers. AB - Cholesterol (Chol) in phosphatidylcholine large unilamellar vesicles (PC LUV) modulated interaction of the bilayers with a class A amphipathic peptide, Ac-18A NH2: Chol increased the peptide binding capacity and reduced the affinity together with the peptide-induced leakage of calcein from LUV. Similar effects of Chol have been observed on the interaction of LUV with apoA-I [Saito, H., Miyako, Y., Handa, T., and Miyajima, K. (1997) J. Lipid Res. 38, 287-294]. Circular dichroism (CD) spectra of the peptide indicated a similar helical structure formation in LUV with and without Chol. The fluorescence spectral shift, quantum yield, anisotropy, and acrylamide-quenching of the peptide Trp indicated that in PC:Chol (3:2) LUV, Ac-18A-NH2 was located in a more polar membrane environment with increased motional freedom and greater accessibility to the aqueous medium. Fluorescence energy transfer from the Trp indole ring to acceptors situated at different depths in the bilayers revealed that the amphipathic peptide penetrated the hydrophobic interior of PC bilayers, while the peptide was located at the polar zwitterionic surface in PC:Chol LUV. The inclusion of Chol causes the headgroup separation of PC at the surface of LUV and increases the binding maximum of the wedge-shaped amphipathic peptide without disrupting the membrane structure. In addition, the rigidifying effect of Chol on PC acyl chains prevents the penetration of the peptide into the bilayer interior. These findings imply that Chol in membranes affects the binding and motional freedom of exchangeable plasma apolipoproteins containing class A amphipathic sequences, e.g., apoA-I and apoCs. PMID- 11900562 TI - Surgical pathology extradepartmental consultation practices. AB - OBJECTIVES: To document the practice parameters and case characteristics associated with personal (expert) consultations. We also examine the value, level of participant (customer) satisfaction, turnaround time, and rate of personal consultations. DESIGN: We asked participants in the College of American Pathologists' Q-Probes program to document cases sent for consultation during 4 months or up to 20 cases. They documented patient and specimen characteristics, the turnaround times, and the participants' levels of satisfaction with the consultation experience. PARTICIPANTS: One hundred eighty laboratories/surgical pathology practices. One hundred seventy-two (95.6%) were from the United States; the remainder were located in Canada and Australia. MAIN OUTCOME MEASURES: Rate and turnaround time of consultations and participant level of satisfaction. RESULTS: A total of 2746 consultation cases were examined for an aggregate consultation rate of 0.5% (median, 0.7%). Institutions with a higher occupied bed size and a greater number of surgical pathology cases both had lower consultation rates (P < or =.05). The median turnaround time (defined as the interval from the date on which the case was sent to the date on which the diagnosis was received) was 6 days. Twenty-nine percent and 68% of cases had a turnaround time within 3 and 7 days, respectively. Fifty-two percent of cases were sent to nationally known experts, and 32% were sent to local experts. Skin (18.0%), hematolymphoid (11.6%), and breast (9.6%) specimens were most commonly sent for consultation. In 70.5% of cases, the consultant confirmed the referring pathologist's original diagnosis, but in 15.9% of cases, the consultant also added significant information. Satisfaction rates were higher with faster turnaround times and verbal reporting. Satisfaction rates were lower for cases in which the patient or the clinician requested the consultation and in which the consultant's diagnosis was ambiguous. CONCLUSIONS: This study establishes a multi-institutional consultation rate of 0.5%, defines the nature of surgical pathology consultations, and demonstrates that satisfaction with consultations is associated with a faster turnaround time and receipt of additional, clinically meaningful information. PMID- 11900561 TI - Further comment on abnormally pigmented organs presenting at autopsy. PMID- 11900563 TI - Xanthochromia. AB - CONTEXT: Diagnosis of subarachnoid hemorrhage is usually established by computed tomographic scanning; however, in a few patients, lumbar puncture to examine the cerebrospinal fluid for erythrocytes and xanthochromia is necessary. Some authorities recommend delaying lumbar puncture by 12 hours following onset of symptoms to ensure sufficient time has elapsed for xanthochromia to develop. This recommendation is based on measuring xanthochromia by spectrophotometry. Our hypothesis was that very few hospital laboratories in the United States use this method. OBJECTIVE: To determine the percentage of hospital clinical laboratories that measure for xanthochromia using spectrophotometry. DESIGN, SETTING, AND PARTICIPANTS: Mail survey to 3500 hospital clinical laboratory directors (in collaboration with the College of American Pathologists). Surveys were mailed in January 2001 and the results tabulated 1 month later. Participation was voluntary. MAIN OUTCOME MEASURES: Percentage of hospital clinical laboratories that use spectrophotometry versus visual inspection. RESULTS: Of the 3500 laboratories surveyed, 2551 (72.9%) responded. Of these, 1944 (76.2%) indicated that they evaluated for xanthochromia. Of the 1952 laboratories that answered the question "How do you report your results?" 1947 (99.7%) reported using visual inspection. CONCLUSIONS: When evaluating for xanthochromia in cerebrospinal fluid, nearly all hospital clinical laboratories in the United States use visual inspection. Given this current reality, the recommendation of delaying lumbar puncture by 12 hours needs to be reassessed. PMID- 11900564 TI - Outpatient phlebotomy success and reasons for specimen rejection. AB - OBJECTIVES: To determine the rate with which blood collection is successful on the initial phlebotomy encounter, the rate with which laboratory personnel judge specimens unsuitable for analysis, and the practice characteristics associated with fewer unsuccessful collections and fewer rejected specimens. DESIGN: Clinical laboratories participating in the College of American Pathologists Q Probes laboratory improvement program prospectively characterized the outcome of outpatient phlebotomies for 3 months or until 20 unsuccessful phlebotomy encounters occurred. By questionnaire, participants provided information about test ordering, patient preparation, and specimen collection. SETTING AND PARTICIPANTS: Institutions in the United States (n = 202), Canada (n = 4), Australia (n = 3), and South Korea (n = 1). MAIN OUTCOME MEASURES: Percentage of successful encounters and percentage of unsuitable specimens. RESULTS: Of 833289 encounters, 829723 were successful. Phlebotomies were unsuccessful because patients were not fasting as directed (32.2%), phlebotomy orders were missing information (22.5%), patients specimens were difficult to draw (13.0%), patients left the collection area before specimens were collected (11.8%), patients were improperly prepared for reasons other than fasting (6.3%), patients presented at the wrong time (3.1%), or for other reasons (11.8%). Only 2153 specimens (0.3%) were unsuitable; these samples were hemolyzed (18.1%), of insufficient quantity (16.0%), clotted (13.4%), lost or not received in the laboratory (11.5%), inadequately labeled (5.8%), at variance with previous or expected results (4.8%), or unacceptable for other reasons (31.1%). Facilities staffed by laboratory-administered phlebotomists reported higher success rates than facilities staffed by nonlaboratory-administered phlebotomists (P =.002). CONCLUSIONS: Most outpatient phlebotomy encounters are successful and result in specimens suitable for laboratory analysis. PMID- 11900566 TI - Epithelial-type and neural-type cadherin expression in malignant noncarcinomatous neoplasms with epithelioid features that involve the soft tissues. AB - CONTEXT: Transmembrane adhesion molecules, epithelial-type cadherin (ECAD) and neural-type cadherin (NCAD), help in regulating transformations between epithelial and mesenchymal cells in the developing embryo and in maintaining the epithelioid phenotype. Consequently, the presence of epithelioid cells in certain malignant noncarcinomatous neoplasms raises speculation that the expression of ECAD and NCAD in these neoplasms may have diagnostic significance. OBJECTIVE: To investigate the utility of ECAD and NCAD immunoexpression in distinguishing malignant (noncarcinomatous) neoplasms with epithelioid features that involve the soft tissues. DESIGN: Membranous immunoreactivity of anti-ECAD and anti-NCAD was evaluated on archived cases selected from the files of the Armed Forces Institute of Pathology. RESULTS: Epithelial-type cadherin was found in biphasic synovial sarcoma (35 of 35 cases), malignant melanoma (13/21), monophasic fibrous synovial sarcoma (13/26), clear cell sarcoma (4/9), poorly differentiated synovial sarcoma (3/13), diffuse mesothelioma (4/20), malignant epithelioid peripheral nerve sheath tumor (1/6), and epithelioid sarcoma (5/62). Neural-type cadherin was observed in chordoma (11/11), biphasic synovial sarcoma (30/35), diffuse mesothelioma (14/20), malignant melanoma (14/25), epithelioid sarcoma (24/63), epithelioid angiosarcoma (1/4), poorly differentiated synovial sarcoma (2/13), clear cell sarcoma (1/10), and monophasic fibrous synovial sarcoma (1/26). Eighteen cases of primary cutaneous squamous cell carcinomas all tested positive for ECAD, whereas NCAD was focally observed in 5 cases. No expression of either molecule was observed in cases of epithelioid hemangioendothelioma (n = 9), alveolar soft part sarcoma (n = 8), and extraskeletal myxoid chondrosarcoma (n = 7). CONCLUSIONS: Epithelial-type and neural-type cadherins are found in a variety of noncarcinomatous neoplasms with epithelioid features that involve the soft tissues and can be utilized, in association with other immunomarkers, in distinguishing chordoma (100% NCAD) from extraskeletal myxoid chondrosarcoma and conventional chondrosarcoma of bone (0% NCAD), squamous cell carcinoma (100% ECAD) from epithelioid sarcoma (8% ECAD), and biphasic synovial sarcoma (100% ECAD) from diffuse mesothelioma (20% ECAD). PMID- 11900565 TI - Estimation of size of clonal unit for keratinocytes in normal human skin. AB - OBJECTIVE: It has been suggested that keratinocyte (KC) stem cells reside at the epicenter of a clonal population of cells. To estimate the territory or surface area covered by a single stem-cell-derived KC population in human skin, clonal skin maps were created from 3 healthy adult women and from normal skin of a psoriatic patient. DESIGN: Two hundred fifty-eight punch biopsy samples of various sizes (ranging from 2 to 8 mm in diameter) were analyzed for clonality employing X chromosome inactivation patterns at the human androgen receptor gene (HUMARA) locus. DNA was isolated and clonality established by significant decrease of either maternal or paternal X chromosome band patterns following restriction enzyme digestion, polymerase chain reaction amplification, and gel electrophoresis. RESULTS: Fifty-three (41%) of 128 two-mm biopsies were clonal, whereas only 6 (14%) of 43 three-mm, 5 (14%) of 36 four-mm, and 3 (8%) of 35 five mm biopsies revealed a clonal population of KCs. By contrast, in 5 different biopsies from a psoriatic patient, including 4- or 5-mm sizes, all but 1 were clonal; even an 8-mm biopsy contained a clonal population of KCs. Mantel-Haenszel chi(2) analysis revealed a P value of.001, reflecting a strong trend in probability for presence of a single clone of KCs as related to size of the biopsy sample. By sequentially analyzing 30 contiguous 2-mm biopsy samples within a given strip of skin, 10 clonal domain changes, as reflected in maternal versus paternal switches, were observed. CONCLUSIONS: These results provide direct evidence of a clonal population of KCs in normal and psoriatic lesion-free skin, and indicate that a clonal epidermal unit of KCs frequently can be detected in small biopsies (2 mm), but that in normal skin sampling, overlapping clones are apparently present in larger (ie, 4-5-mm) biopsies, producing nonclonal patterns. The clonal domain of progeny in normal skin has a rather limited territorial boundary (2 mm in diameter). However, in lesion-free skin from a psoriatic patient, there may be clonal expansion of KCs due to perturbation in epidermopoiesis and/or stem cell distribution. PMID- 11900567 TI - Glial cell-derived neurotrophic factor expression in normal human lung and congenital cystic adenomatoid malformation. AB - CONTEXT: It has been recently suggested that dysregulation of developmental factors and disruption of cell turnover could play a role in the pathogenesis of congenital cystic adenomatoid malformation of the lung (CCAM). The glial cell derived neurotrophic factor (GDNF) is a growth factor involved in organogenesis, and the temporal pattern of GDNF expression suggests that this factor may play a role in lung development. DESIGN: We studied GDNF expression by immunohistochemistry in postnatally resected CCAM of the lung (n = 10), normal fetal lung (n = 5), and normal postnatal lung (n = 5). We also studied the association between GDNF expression and both cell proliferation and apoptosis. RESULTS: GDNF was expressed in both epithelial and endothelial compartments of normal fetal lung, whereas no expression was found in normal postnatal lung. In contrast, in CCAM tissue, there was strong GDNF immunostaining that was restricted to epithelial cells. The percentage of proliferating epithelial cells was higher in CCAM tissue than in normal postnatal lung (6.3% vs 1.7%, P <.005). Apoptotic bodies were found in the mesenchyme of both normal fetal lung and CCAM tissue, whereas virtually no apoptotic bodies were detected in normal postnatal lung. CONCLUSIONS: Abnormal GDNF expression in CCAM suggests a dysregulation of the GDNF signaling pathway and argues in favor of a focal arrest in maturation during development. GDNF expression in lung tissue seems to be correlated with cell proliferation, suggesting that this factor could play a role in the growth of both fetal lung and CCAM. PMID- 11900568 TI - Clonality of combined tumors. AB - CONTEXT: Tumors with mixed morphologic patterns (combined tumors) are sometimes encountered, and questions often arise regarding the mechanism of molecular pathogenesis of each component and their relationships. OBJECTIVE: To determine whether different components of combined tumors contain the same or different genetic alterations, thus providing evidence for their clonality. MATERIALS AND METHODS: Six combined tumors with 2 components (in each case, both components showed epithelial differentiation morphologically) were studied by microdissecting tumor cells from each morphologic area followed by loss of heterozygosity analysis. RESULTS: In 1 of the cases studied, the different morphologic areas contained different patterns of genetic alterations. In the remaining 5 cases, the different morphologic areas harbored identical genetic changes in the chromosome regions studied. The latter group, interestingly, included a colonic tumor with an area of tubulovillous adenoma and an area of neuroendocrine carcinoma, and 2 lung tumors with squamous carcinoma and small cell carcinoma components. CONCLUSIONS: Our results suggest that in the majority of combined tumors, cells with different phenotypes share similar genotype and may arise from a single precursor cell. However, in a minority of these tumors, different areas may be derived from different precursor cells. PMID- 11900569 TI - Pathologists in a teaching institution assess the value of the autopsy. AB - CONTEXT: With the advent of modern diagnostic technology, use of the autopsy as a means of assessing diagnostic accuracy has declined precipitously. Interestingly, during the same period, the rate of misdiagnosis found at autopsy has not changed. OBJECTIVES: To ascertain why an autopsy was requested, whether or not questions asked by clinicians were specifically addressed, and what types of misdiagnoses were found. DESIGN: One hundred forty-two consecutive autopsy records from the University of Arkansas for Medical Sciences Hospital were reviewed. In the same period, 715 deaths occurred, giving an overall autopsy rate of 20.14%. RESULTS: Of the 125 autopsies in which the problem-oriented autopsy request was available for review, a reason for the autopsy was given in only 69 cases (55%). One hundred three clinical questions were asked, and of these, 81 were specifically addressed in the final anatomic diagnosis, 10 were addressed in some part of the autopsy report but not in the final anatomic diagnosis, 10 were not addressed at all, and 2 could not be answered by the autopsy. Sixty-one autopsies revealed 81 misdiagnoses: 47 class I (missed major diagnosis that, if detected before death, could have led to a change in management that might have resulted in cure or prolonged survival) and 34 class II misdiagnoses (missed major diagnosis in which antemortem detection would have not led to a change in management). CONCLUSIONS: The autopsy continues to be a vital part of medical education and quality assurance. It is important for the clinician to provide a clinical summary and specific clinical questions to be addressed or to speak directly with the pathologist and for the pathologist to provide answers that are easily accessible within the autopsy report. In this way, a problem-oriented autopsy can be performed based on questions raised by the clinician and the pathologist as a result of the gross dissection and microscopic evaluation. PMID- 11900570 TI - Coronary angiographic findings in patients with clinical unstable angina according to cardiac troponin I and T concentrations in serum. AB - CONTEXT: Elevated cardiac troponin levels have been reported to identify unstable angina patients at high risk. OBJECTIVE: To examine the relation of cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels to findings of coronary angiography in these patients. METHODS: Samples for troponin estimation were taken every 4 hours throughout the first 48 hours after admission before angiography in 34 patients with primary unstable angina. Patients were considered to be troponin positive if the marker was increased (>0.04 microg/L for cTnT and >0.03 microg/L for cTnI) in at least one sample collected. RESULTS: An increased troponin (I or T) concentration was documented in 14 patients (41.2%). Twelve patients (35.3%) had elevations of both markers, whereas the remaining 2 patients had elevations of cTnI or cTnT alone. Patients with or without increased troponin levels did not differ with respect to degree of coronary disease at angiography. However, patients with elevated troponin concentrations had more complex lesion characteristics. In 69% of patients with increased cTnI levels and in 77% of patients with increased cTnT levels, type B2 or C lesions were documented with presence of ulcerated plaques and thrombus formation. In contrast, only 23% of the patients with elevated cTnI or cTnT levels had type A lesions compared with 71% of patients with negative troponin concentrations. CONCLUSIONS: Patients with unstable angina who have significant release of cTnI and/or cTnT have evidence of more complex lesions on coronary angiography, supporting the hypothesis that both troponins might be used without distinction as surrogate markers for microembolization from thrombus formation on a disrupted plaque. PMID- 11900571 TI - p53 protein overexpression in bone marrow biopsies of patients with Shwachman Diamond syndrome has a prevalence similar to that of patients with refractory anemia. AB - CONTEXT: Shwachman-Diamond syndrome (SDS) is a rare inherited disorder characterized by pancreatic insufficiency, neutropenia, and in some patients, metaphyseal dysostosis. Patients with SDS are at a high risk for development of bone marrow failure, myelodysplastic syndrome, and acute leukemia. The p53 gene plays a major role in cell-cycle regulation, particularly in the presence of a genetic alteration in DNA, a critical step for the initiation of leukemogenesis. p53 gene up-regulation and p53 protein overexpression may occur as a cellular reaction to significant DNA damage. Shwachman-Diamond syndrome and refractory anemia patients have close similarities in the prevalence of acute leukemia and in cell-cycle changes in bone marrow cells. This similarity was further investigated for p53 protein overexpression using archived tissue from patients with hematologic diseases having various leukemic propensities, including SDS and refractory anemia. METHODS: Immunohistochemical staining for p53 protein overexpression was performed on bone marrow biopsies from 9 patients with SDS. These specimens were compared with biopsies from 71 patients with acquired hematologic disorders with variable risk levels for leukemia, including acquired aplastic anemia (n = 14), refractory anemia (n = 46), and various acquired cytopenias (n = 11), as well as 37 control subjects. RESULTS: p53 protein overexpression was identified only in patients with SDS and in patients with refractory anemia; these groups exhibited comparable prevalences of 78% and 72%, respectively. None of the patients with acquired aplastic anemia, acquired cytopenias, or in the control group showed overexpression of p53 protein. CONCLUSION: The prevalence of p53 protein overexpression in SDS is significantly different from that in acquired aplastic anemia and acquired cytopenias, but it is similar to the prevalence in refractory anemia. We speculate that p53 protein overexpression in this bone marrow failure syndrome may represent an early indicator of significant DNA genetic alteration, which is a crucial step in the process of leukemogenesis. PMID- 11900572 TI - Validation of p53 immunohistochemistry as a prognostic factor in breast cancer in clinical practice. AB - CONTEXT: Abnormal p53 tumor suppressor gene expression as detected by immunohistochemistry is a possible prognostic factor in breast cancer. The difference in techniques used to evaluate the expression of mutated p53 protein is under intense scrutiny, as well as its uses either independently or in conjunction with other prognostic factors in breast cancer. OBJECTIVE: To determine whether p53 immunohistochemistry can be used as a reliable indicator of the presence of mutated nuclear p53 protein, and whether this method can be performed reliably in a community hospital's clinical practice. DESIGN: ne hundred twenty-two cases of breast carcinoma were stained and analyzed for the presence of p53 protein using DO-7 (Dako Corporation, Carpinteria, Calif) p53 antibody. RESULTS: Of the 122 cases of invasive carcinoma studied, 23 (18.7%) were positive for p53, and 16 (16.3%) of 98 cases with coexisting ductal carcinoma in situ were positive for p53. This finding is in agreement with comparable published studies. CONCLUSIONS: Based on the results of this study, we conclude that p53 immunohistochemistry qualifies as a diagnostic technique suitable for clinical practice in a community hospital. Its detection may be particularly promising in clinical trials of new molecular therapies directed at the p53 tumor suppressor gene. PMID- 11900573 TI - Expression of cyclin D1 in normal, metaplastic, hyperplastic endometrium and endometrioid carcinoma suggests a role in endometrial carcinogenesis. AB - CONTEXT: Endometrioid carcinoma is often preceded by characteristic histopathologic lesions known as endometrial hyperplasia. Estrogen appears to be involved in the development of endometrioid carcinoma. Other mechanisms of endometrial carcinogenesis include mutations in p53 and PTEN tumor suppressor genes and overexpression of cyclin D1. However, the pattern of cyclin D1 expression is not well defined in normal, hyperplastic, neoplastic, and metaplastic endometrium. DESIGN: Cyclin D1 immunohistochemical analysis was used to evaluate 108 fixed, paraffin-embedded endometrial biopsy specimens and uterine resections obtained from 108 patients. Specimens included proliferative and secretory endometria, simple and complex hyperplastic lesions, and endometrioid adenocarcinoma. Normal and metaplastic surface epithelia were also evaluated independently of glandular morphologic features. RESULTS: Cyclin D1 was significantly overexpressed in glands with complex hyperplasia and endometrioid adenocarcinoma compared with proliferative or secretory endometrium and simple hyperplasia. Significant overexpression was also noted in papillary, syncytial, and squamous metaplasias compared with normal surface epithelium or epithelium with tubal metaplasia. CONCLUSION: Overexpression of cyclin D1 increases from normal endometrium to hyperplasia and carcinoma, suggesting that it may play a role in endometrial carcinogenesis. Overexpression of cyclin D1 in endometrial glands was independent from overexpression of cyclin D1 in surface metaplastic epithelium. PMID- 11900574 TI - Gastric endocrine pancreatic heterotopia. AB - Heterotopic pancreas is a relatively infrequent lesion most often found in the stomach. Four histologic types are recognized: total, canalicular, exocrine, and endocrine heterotopia. To our knowledge, only 2 cases of purely endocrine heterotopic pancreas have been reported in detail. We describe the case of a patient with gastric and duodenal ulcers and gastric endocrine heterotopia. The lack of mass formation, histomorphology, and immunohistochemical features simulating islets of Langerhans supported the diagnosis. We conclude that purely endocrine heterotopic pancreas is a very rare entity that, when present, can simulate a primary or metastatic neuroendocrine tumor. Adequate sampling of the specimen, histomorphologic pattern, and immunohistochemistry are important for the purpose of distinguishing between a neuroendocrine tumor and purely endocrine pancreatic heterotopia. PMID- 11900576 TI - American cutaneous leishmaniasis. AB - We present 3 cases of American cutaneous leishmaniasis occurring in soldiers of a unit of US Army Rangers who parachuted into the jungles of Panama. Shortly after returning to the United States, these 3 soldiers each developed a crusted, indurated papule, which slowly enlarged during the following 6 weeks. Routine microscopy of skin biopsies revealed a dermal granulomatous inflammation and a predominantly lymphoid infiltrate. Numerous histiocytes contained small oval organisms with bar-shaped paranuclear kinetoplasts, morphologically consistent with leishmanial parasites. Cultures grew Leishmaniasis brasiliensis, subspecies panamensis. The soldiers were treated with intravenous pentavalent antimonial therapy daily for 20 days with good clinical improvement. Epidemics of leishmaniasis occur periodically in tropical regions of the world, and leishmaniasis has emerged in new settings, for example, as an acquired immunodeficiency syndrome-associated opportunistic infection. With an increasingly mobile society, it is important to be familiar with the clinical and histopathologic appearance of conditions such as leishmaniasis, which are common in tropical and subtropical regions and are increasingly significant in other regions of the world. PMID- 11900575 TI - Epithelioid leiomyosarcoma of the external deep soft tissue. AB - Epithelioid leiomyosarcoma in the external deep soft tissue is extremely rare. Most epithelioid leiomyosarcomas occur in the uterus. We present a case of epithelioid leiomyosarcoma occurring in the muscle of the thigh of a 78-year-old man. Histologically, the tumor predominantly consisted of round or polygonal cells arranged in sheets with a focal spindle cell component. Immunohistochemical analysis revealed that the tumor cells expressed vimentin, alpha-smooth muscle actin, and alpha-sarcomeric actin. The tumor was negative for desmin, S100 protein, glial fibrillary acidic protein, pan-keratin, epithelial membrane antigen, CAM 5.2, HMB-45, leukocyte common antigen, factor VIII-associated antigen, and CD34. Electron microscopically, some tumor cells contained abundant actin-type filaments in their cytoplasm. PMID- 11900577 TI - Transient myeloproliferative disorder with erythroid differentiation in Down syndrome. AB - A newborn with a karyotype of 47, XY, +21 presented at birth with a white blood cell count of 27 700/microL of which 61% were blast cells. The blast cell morphologic structure was initially not characteristic of any particular lineage, although the cytoplasm contained fine granules and occasional small vacuoles. Routine cytochemical stains were negative, except one for nonspecific esterase that was faintly positive in most of the blast cells. Flow cytometric analyses showed that the blast cells expressed glycophorin A with a subset dimly coexpressing CD45 and were negative for CD34, CD71, myeloid, lymphoid, and platelet-associated antigens. These immunophenotypic findings were consistent with an abnormal erythroid phenotype. A few days postpartum, markedly dysplastic erythroid precursor cells appeared in the peripheral blood and increased in number as the early blast cells decreased. After a period of subdued blast cell production, a second wave of increase in the number of blast cells and dysplastic erythroblasts followed and ended with the disappearance of circulating abnormal cells. The child is now 5 years old and no major illness has been reported since the remission of this disorder. This case most likely belongs to the category of transient myeloproliferative disorders, although the erythroid-like phenotype of blast cells and the evidence of single-lineage maturation to circulating dysplastic erythroid precursors allow the suggestion that this process could represent a special form of a self-limited hematologic disorder in Down syndrome. PMID- 11900579 TI - Malignant stromal tumor of the gallbladder with interstitial cells of Cajal phenotype. AB - Malignant mesenchymal tumors of the gallbladder are exceedingly rare. We report a malignant stromal tumor of the gallbladder with a phenotype of interstitial cells of Cajal. To our knowledge, only the benign counterpart of this tumor has been described previously. A 34-year-old woman presented with right upper quadrant abdominal pain. At the time of cholecystectomy, the gallbladder was noted to have a thickened wall and a polypoid mass arising in the neck of the gallbladder. Histologic sections showed a cellular proliferation of spindle neoplastic cells that were arranged in short fascicles. Numerous mitotic figures and foci of necrosis were noted. The neoplastic cells expressed CD117 (c-Kit protein) and vimentin. They were negative for smooth muscle actin, desmin, myoglobin, cytokeratin, S100 protein, and CD34. Our case demonstrates that a malignant stromal tumor that is histologically and immunohistochemically identical to gastrointestinal stromal tumor can occur in the gallbladder, and that the expression of CD117 may be of therapeutic importance. PMID- 11900580 TI - Pulmonary abscesses in congenital syphilis. AB - Congenital syphilis remains a public health concern in the United States. Infants whose mothers are treated in the third trimester without adequate prenatal care have an increased risk of morbidity and mortality in the immediate perinatal period. The identification of Treponema pallidum in tissue is definitive confirmation of infection. We report the case of a 32-week gestational age infant born to a mother treated for syphilis 7 days prior to delivery. The infant died 12.5 hours after birth. At autopsy, there was extensive acute hyaline membrane disease. In addition, there were bilateral pulmonary abscesses with spirochetes. The onset of maternal disease was unknown, but was probably early in or prior to the pregnancy. This is an unusual case of pulmonary involvement in congenital syphilis. PMID- 11900578 TI - Fine-needle aspiration of renal angiosarcoma. AB - Angiosarcoma of the kidney is an unusual neoplasm, and primary renal angiosarcoma is exceedingly rare, with fewer than 11 well-documented cases reported to date. To our knowledge, no publication to date has correlated the fine-needle aspiration cytologic findings in renal angiosarcoma with the gross, histologic, and immunohistochemical findings. A 50-year-old man presented with a left kidney mass and multiple liver and pulmonary nodules. Computed tomography-guided fine needle aspiration biopsies of the renal mass and a hepatic nodule were performed and demonstrated malignant spindle cells consistent with angiosarcoma. The diagnosis was confirmed at autopsy through histologic examination and associated ancillary studies. This case presents the fine-needle aspiration cytologic findings in renal angiosarcoma and correlates these findings with the gross pathologic, histologic, and immunohistochemical findings. PMID- 11900582 TI - Pathologic quiz case. A 54-year-old man presents with severe back pain. PMID- 11900581 TI - Intratubular embryonal carcinoma. AB - Although intratubular embryonal carcinoma has been described adjacent to invasive embryonal carcinoma, to our knowledge it has not been reported as an isolated finding. We present in this report the histologic and immunohistochemical findings of 2 cases of intratubular embryonal carcinoma. One case was exclusively intratubular embryonal carcinoma without an invasive component in the same testis. A malignant mixed germ cell tumor in the contralateral testis had been previously excised. The second case is predominantly composed of intratubular embryonal carcinoma adjacent to a malignant mixed germ cell tumor. In one case, the intratubular embryonal carcinoma was immunoreactive for CD30, AE1/AE3, cytokeratin 7 focally, and p53. It was negative for cytokeratin 20, p21, and alpha-fetoprotein. These findings are strongly supportive of the opinion that intratubular embryonal carcinoma is the precursor of invasive embryonal carcinoma. PMID- 11900583 TI - Pathologic quiz case. A sinonasal mass in a 79-year-old African American woman. PMID- 11900584 TI - Pathologic quiz case. Systemic symptoms in a human immunodeficiency virus type 1 positive 30-year-old man. PMID- 11900585 TI - Pathologic quiz case. Intraventricular mass in a 31-year-old man. PMID- 11900586 TI - Laboratory diagnosis of dysfibrinogenemia. AB - Dysfibrinogenemia is a coagulation disorder caused by a variety of structural abnormalities in the fibrinogen molecule that result in abnormal fibrinogen function. It can be inherited or acquired. The inherited form is associated with increased risk of bleeding, thrombosis, or both in the same patient or family. Traditionally, dysfibrinogenemia is diagnosed by abnormal tests of fibrin clot formation; the thrombin time and reptilase time are the screening tests, and the fibrinogen clotting activity-antigen ratio is the confirmatory test. The inherited form is diagnosed by demonstrating similar laboratory test abnormalities in family members, and if necessary by analysis of the fibrinogen protein or fibrinogen genes in the patient. The acquired form is diagnosed by demonstrating abnormal liver function tests and by ruling out dysfibrinogenemia in family members. This article reviews the laboratory testing of dysfibrinogenemia and presents an algorithm for sequential test selection that can be used for diagnosis. PMID- 11900590 TI - The age-related changes in the incidence of 'natural' anti-sperm antibodies suggest they are not auto-/isoantibodies. AB - PROBLEM: Establishing the age-dependent patterns of sperm antibody levels among normal humans. METHODS OF STUDY: Sera samples from 498 healthy subjects aged 0-97 years - 246 males and 252 females - were tested by the gelatin agglutination test of Kibrick, tray agglutination test of Friberg, sperm immobilization test of Isojima and enzyme-linked immunosorbent assay (ELISA). RESULTS: We found a significant increase in the level of sperm agglutinins after 40 years, which decreased after 88 years. The antibodies detected by ELISA were the highest among prepubertal subjects and also declined with aging. No age-dependent changes were established for the sperm immobilizins. With few exceptions, there were no significant differences between male and female sera, as well as between sera of newborn and their mothers. CONCLUSIONS: These data are similar to those established for the age-dependent changes of antibodies towards exogenous antigens, suggesting that the 'naturally occurring' antibodies against human spermatozoa are not auto-/isoantibodies. PMID- 11900591 TI - Normal pregnancy is associated with peripheral leukocyte activation. AB - PROBLEM: Normal pregnancy has been described as both a pro-inflammatory condition and a T helper (Th)2-dominated state. Deviations in the percentage of different subpopulations of circulating leukocytes have been detected, although with conflicting results. This study was designed to analyse further the phenotype of subpopulations of peripheral blood leukocytes in normal pregnant women. METHOD OF STUDY: Whole-blood flow cytometry was used to differentiate subsets of leukocytes using directly labeled monoclonal antibodies to specific cell surface antigens and to a panel of activation-associated markers in 33 normal pregnant women in their third trimester and in 26 non-pregnant controls. RESULTS: We found a significant increase in the proportion of granulocytes and of CD8+ T lymphocytes during pregnancy. Up-regulation of the expression of adhesion molecules was observed on granulocytes, monocytes and T lymphocytes. CONCLUSIONS: Pregnancy alters the representation of leukocyte subpopulations in the maternal circulation and is associated with systemic activation of leukocytes. PMID- 11900592 TI - Immunoglobulins and cytokines level in follicular fluid in relation to etiology of infertility and their relevance to IVF outcome. AB - OBJECTIVE: The aims of the present study were to (i) determine the presence and concentration of albumin fractions (alpha1, alpha2, beta, gamma), immunoglobulins (IgA, IgG, IgM) and cytokines [interleukin (IL)-6, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF)] in periovulatory ovarian follicular fluid (FF) of in vitro fertilization (IVF) patients, (ii) examine the relationship between these parameters and the etiology of infertility as well as the IVF outcome and (iii) find out if these parameters in FF could be used as a predictive factor of IVF outcome. DESIGN: The levels of albumin fractions, immunoglobulin and cytokines were measured from women who underwent IVF therapy for various indications and the results were compared between the patient groups and IVF outcome. MATERIALS AND METHODS: Follicular fluid was obtained from 160 IVF patients. A total of 79 patients underwent controlled ovarian hyperstimulations (COH) either with follicle-stimulating hormone (FSH) or HMG. Whereas, the HMG was used for the second set of patients (n=81) - after down regulation with gondotropin-releasing hormone agonists (Gn-RHa) - the protein fractions were determined using electrophoresis separation. Immunoglobulins were measured using a commercial kits and the concentration of cytokines was determined by the highly sensitive enzyme-linked immunosorbent assay (ELISA) methods. RESULTS: The stimulation regimens used have no effect on albumin (alpha1, alpha2, beta, gamma) and immunoglobulin (IgA, IgG, IgM) concentrations, as no significant difference was observed between the two groups. Besides, no specific relationship was found between the concentration of these investigated parameter in FF and etiology of infertility or fertilization, cleavage and pregnancy rate. Besides, there were no significant differences between the groups for any cytokine investigated. Moreover, there were no correlations between the concentration of IL-6, IL-8 and GM-CSF in FF and steroid hormone concentration in the blood at the day of oocytes retrieval or IVF outcome. IN CONCLUSION: Total protein, albumin fraction, immunoglobulins and cytokines level in FF of patients undergoing COH for IVF therapy for various etiology of infertility could not be a useful parameter for predicting IVF outcome. PMID- 11900594 TI - Changes in the number of polymorphonuclear leukocytes and concentrations of IL-8 and granulocyte elastase in the vaginas of normal pregnant women. AB - PROBLEM: It is unknown whether the number of vaginal polymorphonuclear leukocytes (vPMNs) of healthy pregnant women changes during pregnancy. METHOD OF STUDY: We studied 23 women after 14.7 +/- 3.6 weeks, 37 women after 25.2 +/- 3.1 weeks, and 26 women after 32.9 +/- 2.0 weeks of gestation. Numbers of total vPMNs, viable vPMNs, dead vPMNs, percentage viability of vPMNs, and concentrations of IL-8 and elastase activity were determined in vaginal washings. RESULTS: The vPMNs, including viable and dead vPMNs, significantly increased with advancing gestation from 9.9 +/- 8.6 x 10(5) to 13.1 +/- 13.4 x 10(5), and to 27.1 +/- 26.6 x 10(5) (r=0.315, P < 0.01). Vaginal IL-8 significantly increased with advancing gestation. However, increases in vaginal elastase activity did not occur. CONCLUSIONS: These results suggested that an increase in vPMNs is an early event, and that the increase in vaginal elastase activity is a late event in relation to the ripening of the uterine cervix. PMID- 11900593 TI - Interleukin-12 secretion by peripheral blood mononuclear cells is decreased in normal pregnant subjects and increased in preeclamptic patients. AB - PROBLEM: It has been reported that T-helper (Th) 2 dominance in normal pregnancy shifts to Th1 dominance in preeclampsia. Peripheral blood mononuclear cell (PBMC) production of interleukin (IL)-12, which induce Th1 responses, has not been compared between these clinical states. METHOD OF STUDY: Peripheral blood mononuclear cell from 35 non-pregnant women, 35 healthy pregnant women, 12 mildly preeclamptic patients, and 15 severely preeclamptic patients were cultured for 24 hr. IL-12 secretion was determined by enzyme-linked immunosorbent assay (ELISA). Th1/Th2 ratios in PBMC were determined flow-cytometrically, and the amounts of HLA-DR and CD14 expression on the monocytes were obtained by flow cytometry. RESULTS: Peripheral blood mononuclear cell from healthy pregnant subjects secreted less IL-12 than non-pregnant women. PBMC from severely preeclamptic patients secreted more IL-12 than those from healthy pregnant subjects, while IL 12 secretion in mild preeclampsia resembled secretion in normal pregnancy. Th1/Th2 ratios correlated were positively with IL-12. Increased HLA-DR antigens and reduced CD14 expression, suggesting monocyte activation, were observed in preeclamptic patients, although monocyte counts were unchanged. CONCLUSION: Decreased IL-12 secretion by PBMC may cause Th2 dominance in normal pregnancy, while increased IL-12 secretion by activated monocytes may cause Th1 dominance in preeclampsia. PMID- 11900595 TI - Evolutionary adaptations to pre-eclampsia/eclampsia in humans: low fecundability rate, loss of oestrus, prohibitions of incest and systematic polyandry. AB - Gestational-hypertension/pre-eclampsia occurs in approximately 10% of human pregnancies. This persistent complication of pregnancy has been reported to occur more frequently in couples conceiving very shortly after the beginning of their sexual relationship and/or after a change in paternity. Primipaternity may be the leading cause of pre-eclampsia in women under 30 years of age when genetic susceptibility to cardio-vascular disease has not yet been expressed, especially in women before their twenties, who for the last 40,000 years have perhaps comprised the age group when the majority of parturients classified as Homo sapiens sapiens initiated their reproductive life. In terms of evolution, the prevalence of pre-eclampsia represents a distinct reproductive disadvantage in humans as compared with other mammals. Indeed, pre-eclampsia is a consequence of the defect of the normal human-specific deep endovascular invasion of the trophoblast. The large size of the human fetal brain imposing this deep trophoblastic invasion induced the need for major immunogenetic compromises in terms of paternal-maternal tissue tolerance. The price that mankind has had to pay to adapt to the pre-eclampsia risk is a low fecundability rate and therefore loss of oestrus, possibly a step in the deviation between apes and hominids. Further, pre-eclampsia risk may be a contributing factor leading to the rejection of systematic polyandry in human societies and have influenced prohibition of incest. PMID- 11900597 TI - Apoptosis in the uterus of mice with pregnancy loss. AB - PROBLEM: The mechanisms mediating pregnancy loss induced by various agents are far from being understood. Thus, we investigated the possible involvement of one such mechanism, the apoptotic process, in pregnancy loss induced by lipopolysaccharide (LPS) or cyclophosphamide (CP) as well as the associated changes in the apoptosis-regulating gene products p53 and bcl-2. METHOD OF STUDY: Pregnancy loss was induced by LPS or CP on days 9 or 12 of pregnancy, respectively. LPS- or CP-associated apoptosis was assessed by the TdT mediated dUTP-biotin nick end labeling (TUNEL) method as well as by DNA fragmentation analysis, while p53 or bcl-2 expression was evaluated by immunohistochemistry. RESULTS: Lipopolysaccharide treatment initiated a resorption process that was accompanied by the appearance of apoptotic cells in the uterus, which increased in number by 24 hr after treatment. Induction of pregnancy loss with CP resulted in the appearance of some apoptotic cells in the uterus, reaching a peak at 72 hr after treatment. DNA fragmentation analysis revealed a DNA ladder at 24 hr after LPS as well as 72 hr after CP treatment. Immunohistochemical analysis demonstrated a continuous p53 expression in the uterus of LPS- or CP-treated mice, which was somewhat elevated at the peak of the apoptotic process. On the other hand, bcl-2 expression in LPS-treated mice could be reciprocally correlated with the apoptotic process, appearing only at its initiation or completion, while in CP-treated mice it was continuously expressed except for some elevation at the completion of the apoptotic process. CONCLUSIONS: Our results suggest a possible role for the apoptotic process in mechanisms mediating pregnancy loss and indicate an involvement of p53 and bcl-2 in its regulation. PMID- 11900598 TI - Ethyl glucuronide-a biological marker for recent alcohol consumption. AB - Ethyl glucuronide (EtG) is a non-volatile, water-soluble metabolite of ethanol, with a high storage stability. It can be detected in body fluids, tissues, sweat and hair for an extended time period after the elimination of ethanol from the body. EtG closes the gap between short-term markers for alcohol consumption such as ethanol or methanol and long-term markers for alcohol misuse such as GGT, MCV and CDT. Due to its specific time-frame of detection and its high sensitivity and specificity, EtG is a promising marker for alcohol consumption and for relapse control that enables the therapist to intervene at an early stage of relapsing behaviour. The aim of this review is to give an overview of analytical techniques for the detection of EtG, its clinical use and remaining questions. PMID- 11900596 TI - Induction of transferrin secretion in murine Sertoli cells by FSH and IL-1: the possibility of different mechanism(s) of regulation. AB - In the present study we examined the capacity of interleukin-1 (IL-1) alpha, beta, interleukin-1 receptor antagonist (IL-1ra) and follicle stimulating hormone (FSH) to induce transferrin secretion by Sertoli cells under in vitro conditions. Primary Sertoli cell (SC) cultures from immature mice secreted constitutively transferrin. Stimulation of these cultures with IL-1alpha, IL-1beta significantly increas?d their capacity to secrete transferrin. Addition of IL-1ra to unstimulated SC cultures did not affect their capacity to secrete transferrin. Stimulation of SC cultures with a combination of both IL-1alpha and FSH or IL 1beta and FSH showed additive effect between IL-1 and FSH in their capacity to induce transferrin secretion by these cells. However, stimulation of Sertoli cells with a combination of both IL-1ra and FSH did not affect their capacity to secrete transferrin compared with FSH-stimulated cultures. Our results may suggest the involvement of testicular paracrine/autocrine factors (IL-1) and endocrine (FSH) factors in the regulation of transferrin secretion by SC. This capacity seems to be differently regulated by these factors. Thus, IL-1alpha and beta may directly affect physiological functions of the testis; which may suggest their involvement in the regulation of spermatogenesis and spermiogenesis processes and male fertility. PMID- 11900599 TI - Adaptative response of antioxidant enzymes in different areas of rat brain after repeated d-amphetamine administration. AB - d-Amphetamine has been shown to be a potential brain neurotoxic agent, particularly to dopaminergic neurones. Reactive oxygen species indirectly generated by this drug have been indicated as an important factor in the appearance of neuronal damage but little is known about the adaptations of brain antioxidant systems to its chronic administration. In this study, the activities of several antioxidant enzymes in different areas of rat brain were measured after repeated administration of d-amphetamine sulphate (sc, 20 mg/kg/day, for 14 days), namely glutathione-S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GRed), catalase, and superoxide dismutase (SOD). When compared to a pair-fed control group, d-amphetamine treatment enhanced the activity of GST in hypothalamus to 167%, GPx in striatum to 127%, in nucleus accumbens to 192%, and in medial prefrontal cortex to 139%, GRed in hypothalamus to 139%, as well as catalase in medial prefrontal cortex to 153%. However, the same comparison revealed a decrease in the activity of GRed in medial pre-frontal cortex by 35%. Food restriction itself reduced GRed activity by 49% and enhanced catalase activity to 271% in nucleus accumbens. The modifications observed for the measured antioxidant enzymes reveal that oxidative stress probably plays a role in the deleterious effects of this drug in CNS and that, in general, the brain areas studied underwent adaptations which provided protection against the continuous administration of the drug. PMID- 11900600 TI - Diamorphine treatment for opiate dependence: putative markers of concomitant heroin misuse. AB - The supply of substitute opioid medication as a treatment for heroin dependence is now common practice. There is growing international interest in the prescription of injectable diamorphine for subgroups of patients who are unable to stop injecting opiate drugs; in the United Kingdom it is estimated that there are currently 300 patients prescribed diamorphine for this purpose. The detection of illicit heroin misuse (through urinary diamorphine metabolites) is confounded in subjects prescribed diamorphine. We investigated the potential to distinguish between the use of street heroin and pharmaceutical diamorphine through the detection in urine of various opiate alkaloids originating in the opium poppy, Papaver somniferum. Over a 7-week period, 532 clients of an urban substance misuse service provided a total of 1122 urine samples for clinical purposes. Using a novel mixed-mode solid-phase extraction and gas chromatography/mass spectrometry technique, we screened samples for morphine, 6-monoacetylmorphine, codeine, meconine, papaverine, noscapine, thebaine and their metabolites. All urine samples from diamorphine-treated patients were positive for morphine. Of samples from patients receiving other treatments, 30% (95%CI: 27-33%) were positive for morphine, indicating probable street heroin misuse. Of morphine positive samples, 61% (95%CI: 55-67%), from the "other treatments" group were positive for at least one of codeine, meconine and putative noscapine or papaverine metabolites. This was reduced to 56% (95%CI: 50-62%) when excluding codeine. Only one sample (0.1%) was positive for any one of these putative markers in the absence of morphine, when excluding codeine. These findings show that the detection of urinary noscapine and papaverine metabolites is useful in distinguishing between use of pharmaceutical diamorphine and street heroin. This may be of benefit to promote safer and more effective prescribing of diamorphine in opiate dependency, and as an outcome measure in trials of diamorphine prescribing. PMID- 11900601 TI - Association between catechol-O-methyltransferase (COMT) polymorphism and severe alcoholic withdrawal symptoms in male Japanese alcoholics. AB - Catechol-O-methyltransferase (COMT) plays a crucial role in central dopaminergic systems. The allelic association of a functional polymorphism of the COMT gene with the onset, course and clinical characteristics of alcoholism in 91 male Japanese alcoholics and 114 male Japanese controls was examined. There were no statistically significant differences in the frequency of the allele between the alcoholics and the controls. Visual disturbances and auditory disturbances among alcoholic withdrawal symptoms were, however, significantly different among the three groups (p < 0.05). The present results suggest that COMT activity could partially effect the phenotype of alcoholism, especially the appearance of delirium tremens, in these subjects. PMID- 11900602 TI - Information processing deficits in withdrawing alcoholics. AB - Prepulse inhibition (PPI) of the acoustic startle response (a reduction in response to an intense, startling stimulus (the pulse) if preceded by 30-150 ms by a weaker, non-startling stimulus) is an established model to index information processing deficits in thought-disordered schizophrenic patients. The present study aimed to investigate the influence of alcohol withdrawal on the PPI effect. Eight withdrawing alcoholic patients underwent testing for PPI of the acoustic startle response (defined as percentage reduction of the response over pulse alone stimulus; prepulses 15 dB above the background) on three occasions (1, 3 and 7 days following the last drink). The results demonstrated remarkably low levels of PPI on days 1 and 3, with this being very robust in three patients who had a history of delirium tremens; there was a trend towards normalization of PPI on day 7. This study, although preliminary, suggests strongly that there is a deficit in the filtering of sensory information in alcohol-dependent patients undergoing alcohol withdrawal. This was most apparent in those with a history of delirium tremens. Further studies are needed to define the cause and chronicity of these deficits. PMID- 11900603 TI - Salivary cortisol measurements during a medically assisted alcohol withdrawal. AB - Previous studies using plasma cortisol estimations have suggested that hypothalmo pituitary-axis (HPA) activation occurs in alcohol-dependent patients during alcohol withdrawal. The present study set out to confirm this finding using salivary cortisol assays, which are a better indicator of plasma free cortisol, the fraction which exerts its physiological effects. Nine alcohol dependent patients provided four saliva samples (at 10 a.m., 2 p.m., 6 p.m. and 10 p.m.) on days 1, 3 and 7 of a medically assisted alcohol withdrawal (corresponding to 1, 3 and 7 days following the last drink, respectively).Withdrawal symptom severity, craving and mood disturbance were also measured. A group of non-alcohol-dependent individuals, without psychiatric or medical disorder, gave four samples at the same times on one day only. Mean daily cortisol levels in our alcohol-dependent population, as calculated by the area under the curve (AUC), decreased significantly over time (mean AUC (nmol/l/hour) on day 1 = 149, on day 7 = 85.7, p = 0.009) and were significantly higher than controls on each day (mean AUC in controls = 28.3, p = 0.001). The cortisol response showed a similar temporal trend to withdrawal symptom severity and mood disturbance. This is consistent with previous studies measuring plasma cortisol in alcohol withdrawal. However, the magnitude of the effect in our study was greater, and in contrast to some previous studies, levels were far from normal by day 7. The comparatively low cortisol response in our one mildly dependent patient suggests that there may be a relationship between dependence severity and the size of the cortisol response to withdrawal. Salivary cortisol sampling could prove to be a useful prognostic tool, with implications for subsequent withdrawal symptom severity, mood disturbances, risk of relapse and alcohol-related cognitive decline. There are implications for developing new treatments for alcohol withdrawal but more studies are needed. PMID- 11900604 TI - Chronic high-dose nitrazepam dependence 123I-IMP SPECT and EEG studies. AB - A patient who took 50-100 mg nitrazepam per day for 25 years is presented. (123)I IMP SPECT (autoradiography method) and EEG were performed sequentially on the subject during and after the withdrawal syndrome. Severe hypoperfusion of the whole brain on SPECT and diffuse slow activity on EEG were demonstrated during the withdrawal syndrome and subsequently remarkably improved. However, the hypofrontal pattern on both early and delayed images in SPECT was unchanged. The changes observed on SPECT and EEG reflect the pathophysiology of dependence and withdrawal. Additionally, the hypofrontal pattern remained unchanged suggesting that organic brain damage can develop as a result of chronic high-dose benzodiazepine abuse. PMID- 11900606 TI - Society for the Study of Addiction: The nature and treatment of dependence. AB - The following are selected and edited abstracts from the Society for the Study of Addiction Annual Symposium, 2-3 November 2000, Leeds, UK PMID- 11900605 TI - Propofol dependency after treatment of tension headache. AB - Propofol is a widely used general anaesthetic with growing usage outside anaesthesia. Despite its potential for abuse only five cases of propofol dependency have yet been reported. These comprise health care professionals with easy access to the drug. This is the first reported case of propofol dependency in a lay person. One should be aware of the abuse potential of propofol with regard to its growing use outside the context of general anaesthesia. PMID- 11900607 TI - A reversibly immortalized human hepatocyte cell line as a source of hepatocyte based biological support. AB - The application of hepatocyte transplantation (HTX) is increasingly envisioned for temporary metabolic support during acute liver failure and provision of specific liver functions in inherited liver-based metabolic diseases. Compared with whole liver transplantation, HTX is a technically simple procedure and hepatocytes can be cryopreserved for future use. A major limitation of this form of therapy in humans is the worldwide shortage of human livers for isolating an adequate number of transplantable human hepatocyes when needed. Furthermore, the numbers of donor livers available for hepatocyte isolation is limited by competition for their use in whole organ transplantation. Considering the cost of hepatocyte isolation and the need for immediate preparation of consistent and functional cells, it is unlikely that human hepatocytes can be obtained on such a scale to treat a large number of patients with falling liver functions. The utilization of xenogenic hepatocytes will result in additional concerns regarding transmission of infectious pathogens and immunological and physiological incompatibilities between animals and humans. An attractive alternative to primary human hepatocytes is the use of tightly regulated human hepatocyte cell lines. Such cell lines can provide the advantages of unlimited availability, sterility and uniformity. We describe here methods for creating transplantable human hepatocyte cell lines using currently available cell cultures and gene transfer technology. PMID- 11900608 TI - Liver transplantation for alcoholic liver disease. AB - Although increasing numbers of alcoholic patients are being referred to liver transplant centres, liver transplantation for alcoholic liver disease still remains controversial, essentially because we are in an era of organ shortage. In fact, the main issue is the likelihood of relapse and its influence on outcome, because it is the possibility of returning to alcohol use that separates patients with alcoholic liver disease from those with other forms of chronic liver disease. In all proposed clinical guidelines of indications for referral and assessment for liver transplantation for alcoholic liver disease, the authors emphasize the risk of alcoholism recurrence and, thus, a multidisciplinary approach is required to select patients who are likely to comply with follow-up and not return to a damaging pattern of alcohol consumption after transplantation. It emerges from all clinical studies that when we take into account the usual criteria of success for liver transplantation, i.e. patient and graft survival, rejection rate and infection rate, alcoholic liver disease is a good indication for liver transplantation. Predictive factors for alcoholic relapse after liver transplantation have been assessed in numerous studies, often with contradictory results making these difficult to analyse and compare. Several predictive factors for alcoholic relapse have been studied: length of abstinence before transplantation, associated psychiatric problems, social conditions, associated drug addiction, age. Abstinence after transplantation is the goal, but the necessary treatment for alcoholic disease can result in considerable improvement, even when complete abstinence is not achieved. Finally, the good results obtained with liver transplantation for alcoholic liver disease should help us to educate the general population about alcoholic disease. PMID- 11900609 TI - Alcohol and upper gastrointestinal tract cancer: the role of local acetaldehyde production. AB - Alcohol is, together with tobacco smoke, the main cause for upper GI tract cancer in industrialized countries. However, the tumour-promoting effects of alcohol intake are poorly understood and alcohol itself is not carcinogenic in the animal model. There is increasing evidence that alcohol metabolism, rather than the alcohol itself, generates carcinogenic and cell-toxic compounds. Acetaldehyde, first metabolite of ethanol, is highly toxic, mutagenic and carcinogenic. Polymorphisms in the genes coding for enzymes responsible for acetaldehyde accumulation and detoxification have been associated with an increased cancer risk. Acetaldehyde can also be produced in the mucosa and by the physiological microflora. This review summarizes the scientific evidence that alcohol intake leads to a local production of acetaldehyde. It describes the role of the oral microflora, the mucosa and the salivary glands in this process and shows that local acetaldehyde production from ethanol may contribute to the carcinogenesis of alcohol intake in the upper GI tract. PMID- 11900610 TI - Oxidant-antioxidant profiles of platelet rich plasma in smokers. AB - Cigarette smoke is a major risk factor for both coronary heart disease and peripheral vascular disease and has been reported to contain many oxidizing agents that lead to generation of free radicals. In this study, we investigated the levels of lipid peroxides (LPO) and antioxidant vitamins (C and E), total thiol content (t-SH), the activities of superoxide dismutase (SOD), glutathione S transferase (GST) and glutathione peroxidase (GPx) in the platelet-rich plasma (PRP) and plasma of 50 smokers and 30 non-smokers. Total cholesterol (TC), low density-cholesterol (LDL-C), triglyceride (TG) and phospholipid (PL) levels of the plasma were significantly higher (p < 0.001) and high density-cholesterol (HDL-C) levels were significantly lower in smokers (p < 0.001) when compared with non-smokers. In plasma and PRP, LPO levels, GST and SOD activities were found to be increased (p < 0.001) in smokers, whereas GPx activity, vitamin C levels and t SH content were found to be decreased. On the other hand, the levels of vitamin E was unchanged in plasma and PRP. The relationships between plasma levels of lipids, LPO and antioxidant systems were also investigated in both groups. A strong positive correlation was found between TC and Vit E (r = 0.5575; p < 0.001), LPO and PL (r = 0.4270; p < 0.01), LPO and GST (r = 0.3770; p < 0.01) and t-SH and GPx (r = 0.3781; p < 0.01) in smokers. These findings reveal a disturbance of oxidant-antioxidant balance by free radicals present in cigarette smoke, which may cause reduction in platelet hyperreactivity and endothelial dysfunction in smokers. PMID- 11900611 TI - Polymorphisms at the DRD2 locus in early-onset alcohol dependence in the Indian population. AB - The susceptibility to alcohol dependence is probably of polygenic origin. Association studies have attempted to identify possible candidate genes that may contribute to the risk to developing dependence. Severe forms of the alcoholism phenotype have been associated with an increased frequency of the Taq A1 allele at the DRD2 locus. Ethnic stratification and non-comparable phenotype may have contributed to the contradictory results in previous studies. We identified probands, using the Schedules of Assessment of Neuropsychiatry (SCAN) schedule, who had onset of alcohol dependence (ICD-10) before 25 years of age. Family members were interviewed using the Family Interview for Genetic Studies (FIGS) schedule to identify patients who had two first-degree relatives with alcohol dependence. Fifty subjects who fulfilled the criteria were selected for the study. These were compared to a normal population from a similar background. The allele frequencies did not differ between the two groups. The Taq1a polymorphism does not seem to be associated with alcoholism in this group of severely affected, young age of onset probands in the southern Indian population. PMID- 11900612 TI - Serotonin transporter (5-HTT) gene polymorphisms and susceptibility to cocaine dependence among African-American individuals. AB - Studies indicate that the serotonin system, particularly the serotonin transporter (5-HTT), may modulate the central effects of cocaine. We investigated whether a polymorphism in the 5' promotor region (5-HTTLPR) of the 5-HTT gene confers susceptibility to cocaine dependence. One hundred and ninety-seven cocaine-dependent African-American subjects and 101 controls were studied. Polymerase chain reaction based genotyping of a biallelic repeat polymorphism in the 5' promotor region yielded 2 alleles containing 484 (S) and 528 bp (L) repeats, respectively. There were no significant differences between controls of European background (n = 40) and African-American controls (n = 61) in distribution of genotypes (European: LL = 32.5%, LS = 40.0%, SS = 27.5%; African American: LL = 27.9%, LS = 57.4%, SS = 14.7%) (chi(2) = 3.60, df = 2, p = 0.16) or allele frequencies (European: L = 52.5%, S = 47.5%; African-American: L = 56.6%, S = 43.4%) (chi(2) = 2.21, df = 1, p = 0.13). When cocaine patients were compared to an ethnically diverse control group (n = 101), frequencies of the L variant (65.0%) were significantly higher while the S variant (35.0%) was less frequent among cocaine patients compared to controls (L = 53.9%, S = 46.1%) (chi(2) = 6.83, df = 1, p < 0.01). Similarly, there were more cocaine patients with the LL genotype (41.1%) and less with the SS genotype (11.2%) compared to controls (LL = 29.7%, SS = 21.8%) (chi(2) = 7.43, df = 2, p < 0.05). However, after restricting controls to African-American individuals only (n = 61), cocaine subjects and controls did not differ significantly with respect to genotype distribution (chi(2) = 4.24, df = 2, p = 0.12) or allele frequencies (chi(2) = 2.83, df = 1, p = 0.10). In conclusion, although comparisons with a heterogeneous control group indicated a possible association between allelic variants of 5 HTTLPR and cocaine dependence among African-American cocaine subjects, this relationship was not observed when the control group was limited to African American people only. Our findings need to be confirmed on larger samples of ethnically matched individuals. PMID- 11900613 TI - Separate and interactive effects of cocaine and alcohol dependence on brain structures and metabolites: quantitative MRI and proton MR spectroscopic imaging. AB - The effects of chronic cocaine and alcohol abuse on human brain structure and metabolites are not fully known. We studied controls (n = 13) and abstinent subjects dependent on cocaine (8), alcohol (12), and cocaine and alcohol (17) using quantitative MRI and proton MR spectroscopic imaging. Talairach-based techniques yielded tissue and CSF volumes and gray- and white-matter concentrations of N-acetylaspartate (NAA), creatine and choline metabolites in multiple brain regions. Alcohol dependents had lower gray-matter NAA concentrations and more sulcal CSF than non-alcohol dependents throughout the brain. They also had less subcortical gray matter and (regionally) less white matter. Cocaine dependents compared with non-cocaine dependents had higher posterior parietal white-matter creatine concentration. They also had less gray and white matter in the prefrontal lobes and in a region encompassing the temporal lobes and cerebellum. Structural white-matter deficits in cocaine dependents were greater with longer duration of cocaine use. Subjects with concurrent cocaine and alcohol dependence had less prefrontal white matter, especially in the anterior cingulate, than subjects dependent on only one substance. Chronically abused cocaine and alcohol each leave multiple metabolic and structural brain defects after long-term abstinence. Concurrent dependence on both substances may aggravate white-matter structural defects, primarily in frontal brain. PMID- 11900614 TI - Hepatitis B and C in alcohol-dependent patients admitted to a UK alcohol inpatient treatment unit. AB - The prevalence of hepatitis B and C infection was studied in a sample of alcohol dependent patients admitted to a specialist alcohol inpatient unit to identify factors associated with hepatitis B and C infection. Laboratory, clinical and socio-demographic data were collected from 277 admissions over a 3-year period who were tested routinely for markers of hepatitis B and C infection. Of the 275 subjects tested for hepatitis C, 27 (9.8%) were positive to the hepatitis anti HCV IgG antibody. Of the 275 subjects tested for hepatitis B, 30 (10.9%) were positive to the hepatitis anti-HBc IgG antibody. Few differences were found between hepatitis B positive and negative subjects. Hepatitis C positive individuals were more likely than Hepatitis C negative patients to have also been infected with the hepatitis B virus (p < 0.001), to have an unplanned discharge (p < 0.005) and to have ever used cannabis (p < 0.005), cocaine (p < 0.001), amphetamines (p < 0.001) or heroin (p < 0.001). They were also more likely to have a co-morbid antisocial personality disorder (p < 0.001), a lifetime diagnosis of opiate dependence (p < 0.001) and cocaine dependence (p < 0.005), higher serum gamma glutamyl transferase (GGT) levels (p < 0.05) and a lower platelet count (p < 0.05). These findings may help clinicians to identify those alcohol dependent patients with risk factors for hepatitis virus infection. PMID- 11900615 TI - A putative sigma1 receptor antagonist NE-100 attenuates the discriminative stimulus effects of ketamine in rats. AB - Ketamine, one of the dissociative anaesthetic agents, has been shown to produce psychotomimetic effects. It has been well documented that activation of sigma receptors is responsible for the pathogenesis of some psychiatric disorders. In the present study, the effects of NE-100, a putative sigma(1) receptor antagonist, was investigated in rats trained to discriminate between ketamine (5 mg/kg, i.p.) from saline under a fixed-ratio 10 food-reinforced procedure. Here we report for the first time that NE-100 (1 mg/kg) produced a shift to the right in the dose-response curve for ketamine's discriminative stimulus effects. These results suggest that the sigma(1) receptor is, at least in part, involved in the discriminative stimulus effects of ketamine. PMID- 11900616 TI - Alcohol dehydrogenase ADH2-1 and ADH2-2 allelic isoforms in the Russian population correlate with type of alcoholic disease. AB - The frequency ADH2-2 allele in the Moscow urban population and a correlation between the ADH2-2 allele, alcoholic dependence without cirrhosis, symptomatic alcoholic cirrhosis and status on hepatitis B and C infection have been studied. One hundred and twenty-three inhabitants of Moscow (50 healthy donors, 36 patients with alcoholic cirrhosis (subdivided into infected and uninfected by HBV and/or HCV) and 37 patients with alcoholic dependence) of a similar age/sex and drinking pattern have been analysed. The frequency of 41% for ADH2-2 allele is characteristic for an urban Moscow population. This value is intermediate between that found for Asian peoples and for Central and Western Europe. There is a negative correlation between the ADH2-2 allele and alcohol misuse (both alcoholic dependence and alcoholic cirrhosis). This correlation is expressed more in alcoholic dependence. In spite of the possession of the ADH2-2 allele (or genotype ADH2-1/2), alcohol misuse increases the risk of cirrhosis. At the same time, positive status for active hepatitis B, C or combined infection B + C (replication markers HBV-DNA or HCV-RNA) increases the risk for symptomatic alcoholic cirrhosis in alcohol abusing patients, independently of ADH2 genotype. PMID- 11900618 TI - Pharmacokinetics of ethanol: a review of the methodology. AB - This paper reviews current concepts on tools for studying the pharmacokinetics of alcohol. It has been known that ethanol metabolism occurs mainly in the liver via alcohol dehydrogenase and an accessory microsomal pathway. The contribution of each pathway has been examined by administration of metabolic inhibitors. The role of gastric alcohol dehydrogenase in the first-pass effects of ethanol has been speculative and may be relatively low. Some pharmacokinetic approaches with mathematical models have elucidated the role of gastric alcohol dehydorgenase, hepatic alcohol dehydrogenase and cytochrome P450 2E1 in ethanol elimination. The scale-up of ethanol elimination kinetics has enabled extrapolation from animal models to human kinetics. The clarification of the pharmacokinetics of ethanol is very important for estimating the effects of ethanol on biological events. PMID- 11900617 TI - Towards a comprehension concerning the clinical effects of therapy with rapid opioid detoxification. PMID- 11900619 TI - The ubiquitin-proteasome system and its role in ethanol-induced disorders. AB - Some of the most fundamental yet important cellular activities such as cell division and gene expression are controlled by short-lived regulatory proteins. The levels of these proteins are controlled by their rates of degradation. Similarly, protein catabolism plays a crucial role in prolonging cellular life by destroying damaged proteins that are potentially cytotoxic. A major player in these catabolic reactions is the ubiquitin-proteasome system, a novel proteolytic system that has become the primary proteolytic pathway in eukaryotic cells. Ubiquitin-mediated proteolysis is now regarded as the major pathway by which most intracellular proteins are destroyed. Equally important, from a toxicological standpoint, is that the ubiquitin-proteasome system is also widely considered to be a cellular defense mechanism, since it is involved in the removal of damaged proteins generated by adduct formation and oxidative stress. This review describes the history and the components of the ubiquitin-proteasome system, its regulation and its role in pathological states, with the major emphasis on ethanol-induced organ injury. The available literature cited here deals mainly with the effects of ethanol consumption on the ubiquitin-proteasome pathway in the liver. However, since this proteolytic system is an essential pathway in all cells it is an attractive experimental model and therapeutic target in extrahepatic organs such as the brain and heart that are also affected by excessive alcohol consumption. PMID- 11900620 TI - Chronic ethanol intake and ageing effects on cortical and basal forebrain cholinergic parameters: morphometric and biochemical studies. AB - Abstract Twenty-eight Wistar rats treated orally with 20% ethanol solution, were divided into two groups: adult group (n = 19) and aged group (n = 9) consisting of animals aged 4 and 12 months, respectively, at the beginning of the treatment. Neurons from the basal nucleus region were counted and the percentage of choline acetyltransferase-immunoreactive cholinergic neurons was determined in adjacent sections. Acetylcholine release and choline-acetyltransferase activity in the cerebral cortex were assessed in the same animals. Nutritional parameters of the ethanol treated animals were monitored and found to be normal. Chronic exposure to ethanol did not result in global neuronal loss or loss of cholinergic neurons in the basal nucleus region. However, a greater expression of ChAT immunoreactivity in the basal nucleus region and a tendency toward increased ChAT activity in the cerebral cortex of the control and treated aged animals, compared respectively to adult ones, were observed. These findings suggest adaptive changes of the aged rats in response to the possible cholinergic hypofunction, manifested as a decreased release of acetylcholine under stimulated conditions. PMID- 11900621 TI - A Short Alcohol Withdrawal Scale (SAWS): development and psychometric properties. AB - The measurement of alcohol withdrawal symptoms is important for the assessment of the alcohol withdrawal syndrome and for the evaluation of the effectiveness of withdrawal treatment interventions. There continues to be a need for an instrument for the measurement of alcohol withdrawal severity which is short, easy to understand (especially by respondents who may feel anxious, confused or physically ill) and easy to administer (for example, within clinical services with limited time and resources).This paper describes the development and psychometric properties of the 10-item Short Alcohol Withdrawal Scale. The SAWS includes five items which represent psychological symptoms (anxious, confused, restless, miserable, memory problems) which accounted for 47% of the variance. A further five items represent physical symptoms (tremor, nausea, heart pounding, sleep disturbance, sweating) and accounted for 11% of the variance. The procedures leading to the development of the scale are described and results are presented showing that the SAWS has high internal consistency, and good construct and concurrent validity. PMID- 11900622 TI - Serum prolactin and response to treatment among cocaine-dependent individuals. AB - Considerable evidence indicates that dopaminergic mechanisms may modulate the central effects of cocaine. We investigated whether basal serum prolactin, a measure of central dopamine activity, differed between cocaine-dependent subjects and controls, and whether prolactin levels among cocaine patients were related to their response to treatment. Eighty-six African-American cocaine-dependent outpatients and 35 African-American controls were studied. Prolactin concentrations in fasting blood samples were assayed by radioimmunoassay. The outcome measures were: number of negative urine drug screens, retention in treatment, counselor ratings of improvement and discharge status. Chi-square tests and independent t-tests were used for data analyses. The basal prolactin(ng/ml) among cocaine patients (9.12 +/- 4.12) was significantly higher compared to controls (7.14 +/- 3.36) (t = 2.52, p < 0.02). Furthermore, the higher prolactin subjects (median prolactin = 7.71) had significantly fewer negative urine screens (p < 0.05) and received less favorable ratings of improvement by counselors (p < 0.01) compared to the lower prolactin group. However, the two groups did not differ significantly in treatment retention (p = 0.13) or discharge status (p = 0.08). The higher basal prolactin among cocaine patients relative to controls may reflect changes in DA activity among cocaine patients. Moreover, higher prolactin seems to be related negatively with certain measures of response to treatment among cocaine patients. PMID- 11900623 TI - Quality of life, sleep, mood and alcohol consumption: a complex interaction. AB - Socio-demographic information was collected from 52 (45 men, seven women) currently drinking moderately dependent alcohol misusers attending an outpatient clinic in South London for medical assessment and treatment. Assessments at baseline and 12-week follow-up included: (i) Severity of Alcohol Dependence (SADQ) (baseline only), (ii) Hospital Anxiety and Depression Scale (HADS); (iii) Nottingham Health Profile sleep subscale (NHP) and (iv) the EuroQoL (EQ-5D). The main outcome measure was drinking at a "sensible level" at 12-week follow-up as recommended by the Royal College of Psychiatrists. There were significant interactions between sleeping badly, lying awake at night and HADS depression scores. The Health-related Quality of Life (HrQoL) of this group was poor compared to general population norms. Lower EQ-5D index scores were associated with poorer educational attainment and lower EQ-5D Visual Analogue Scale (VAS) patient ratings with greater baseline alcohol consumption. Clinician ratings on the EQ-5D VAS were consistently lower than the patient ratings. The correlations between patient self-assessment and clinician ratings (EQ-5D VAS) were not significant. The forty-seven subjects (90%) who were successfully followed-up showed a significant reduction in the total amount of alcohol consumed. Ten (21%) subjects returned to sensible drinking levels at 12 weeks but there was not a corresponding improvement in HrQoL, sleep, or affective status scores or biochemical measures in these subjects. PMID- 11900624 TI - The relative impact of waiting time and treatment entry on drug and alcohol use. AB - One hundred and twenty-three treatment-seeking substance misusers were recruited to a study assessing the early impact of treatment. Participants were interviewed at treatment entry and 3 and 6 months later, regardless of their treatment status (i.e. including those who had dropped out of treatment), while additional data were obtained from the two assessment interviews carried out prior to the initiation of treatment. Three consistent observations can be applied to both the opiate misuser (n = 61) and problem drinker samples (n = 62): (1) the period of pre-treatment wait (mean of 8 weeks) was characterized by stable patterns of substance misuse with no significant 'spontaneous' improvement in indices of severity of drug or alcohol problems; (2) the period immediately following initiation of treatment was associated with substantial reductions in the quantity and frequency of substance use, an effect not influenced by the length of time for treatment initiation; (3) these benefits are maintained to 6 months after treatment initiation. The waiting period for treatment initiation does not seem to be characterized by significant changes in drug or alcohol use patterns, at least among those who made it into treatment, with clear and sustained improvements irrespective of the length of treatment wait. PMID- 11900625 TI - Proceedings of the Areca Nut Symposium. November 2000, London, United Kingdom. PMID- 11900626 TI - Global epidemiology of areca nut usage. AB - A substantial proportion of the world's population is engaged in chewing areca nut and the habit is endemic throughout the Indian subcontinent, large parts of south Asia and Melanesia. A large variety of ingredients, including tobacco, may be used along with areca nut constituting a betel quid. The composition and method of chewing can vary widely from country to country and these population variations are described in this review. Some populations are known to use areca nut without tobacco providing good opportunities to further research the carcinogenecity of areca nut. Some interesting trends on chewing patterns have emerged from recent data, suggesting a decline in the habit in some countries such as Thailand while the prevalence of areca nut use is rising in India and Taiwan. PMID- 11900627 TI - Anthropological perspectives on use of the areca nut. AB - This paper first reviews the identity of historical and contemporary users of the areca nut. It then considers the reasons given by users for indulging in areca consumption, drawing upon historical, ethnographic and experimental sources of evidence for the effects which users have sought to derive from it. Particularly important is the social context in which consumption occurs, and the social meanings attached to areca use and exchange. Finally, a possible evolutionary hypothesis is postulated to explain the origins of areca use as a form of behaviour indicating reproductive availability. However, diverse culturally explicit reasons underlie usage and what may once have been a sufficient rationale for consumption may no longer justify this pattern of behaviour. PMID- 11900628 TI - Chemical and analytical aspects of areca nut. AB - A capillary zone electrophoretic (CZE) method for the rapid analysis of the major alkaloids (arecoline and guavacoline) in areca nut extract is described. Areca nuts were pulverized and then extracted with water by sonication in a water bath. After centrifugation, the supernatant was analysed on a fused-silica capillary with 100 mM ammonium acetate-acetic acid (pH 4.6) as the running buffer at a voltage of 20 kV and temperature of 30 degrees C. The method is applicable to the analysis of alkaloids in the nut, commercial preparations (pan masala) and in the saliva of areca nut chewers. PMID- 11900629 TI - Metabolic effects of the consumption of Areca catechu. AB - Betel nut (Areca catechu) is chewed regularly by at least 10% of the world population, imported by immigrant users wherever they settle, and is the fourth most widely used addictive substance. It is thought, by users, to soothe the digestion and to be a stimulant and its use has a major role in social situations. Specific arecal alkaloids act as competitive inhibitors of GABA receptors and have widespread effects in the body, including actions on the brain, cardiovascular system, lungs, gut and pancreas. Nitrosated derivatives of arecal alkaloids, proven carcinogens inducing tumours throughout the upper gut and foregut derivatives in animals, are also associated with increased tumour risks in man. These nitrosated compounds are also diabetogenic in CD1 mice, producing a type 2 diabetes with obesity. Increased central obesity is found in association with betel usage in man as well as increases in circulating markers of inflammatory and cardiovascular damage. The effects of chronic betel usage in man are at least as diverse as those of smoking and the habit increases the risks of ill health. PMID- 11900630 TI - Neurological aspects of areca and betel chewing. AB - Betel quid chewing has been claimed to produce a sense of well-being, euphoria, warm sensation of the body, sweating, salivation, palpitation, heightened alertness and increased capacity to work. These effects suggest that betel quid chewing affects predominantly the central and autonomic nervous systems. Several studies have been conducted to elucidate the central and autonomic effects of betel quid chewing. The results are: (1) betel quid chewing increased the heart rate with onset within 2 minutes, maximal effect within 4-6 minutes and an average duration of 16.8 minutes. The cardio-acceleratory response was more prominent for fresh and occasional chewers than for habitual chewers; (2) betel quid chewing increased the skin temperature with onset and duration similar to a cardio-acceleratory response. The hyperthermic effect was abolished by atropine and partly inhibited by propranolol. (3) Betel quid chewing had no effect on simple reaction time but shortened the choice reaction time. (4) Betel quid chewing produced widespread cortical desynchronization of EEG. (5) Chewing of one or two betel quids attenuated the sympathetic skin response while continued consumption of more than two betel quids affected the RR interval variation. (6) Plasma concentrations of noradrenaline and adrenaline were elevated during betel quid chewing. These studies have confirmed several effects claimed by betel quid users. The effects of betel quid chewing appeared to be habit-related and dose dependent. Although arecoline has been thought to be responsible for several effects of betel quid chewing, the present data suggest a role also played by sympathetic activation. PMID- 11900631 TI - The oral health consequences of chewing areca nut. AB - Deleterious effects of areca nut on oral soft tissues are published extensively in the dental literature. Its effects on dental caries and periodontal tissues, two major oral diseases, are less well researched. Areca-induced lichenoid lesions mainly on buccal mucosa or tongue are reported at quid retained sites. In chronic chewers a condition known as betel chewer's mucosa, a discoloured areca nut-encrusted change, is often found where the quid particles are retained. Areca nut chewing is implicated in oral leukoplakia and submucous fibrosis, both of which are potentially malignant in the oral cavity. Oral cancer often arises from such precancerous changes in Asian populations. In 1985 the International Agency for Research on Cancer concluded that there is limited evidence to conclude that areca chewing may directly lead to oral cancer. There is, however, new information linking oral cancer to pan chewing without tobacco, suggesting a strong cancer risk associated with this habit. Public health measures to quit areca use are recommended to control disabling conditions such as submucous fibrosis and oral cancer among Asian populations. PMID- 11900632 TI - Areca nut use following migration and its consequences. AB - Areca nut use is widespread in the Oriental countries, affecting approximately 20% of the world's population. The combined use of areca nut and smokeless tobacco (ST) is practiced particularly in the Indo-Chinese continents. While there is considerable global variation in the use of these products, migrant studies relevant to areca nut use is of considerable interest to epidemiologists in suggesting the extent to which these environment exposures are important in the aetiology of different cancers and other health-related consequences. Studies on Indian migrants to the Malay peninsula, South and east Africa and various Asian ethnic groups resident in several parts of the United Kingdom have shown that the consumption of areca nut (often mixed with ST) is highly prevalent in these communities. Available data on the prevalence of areca chewing among these migrant populations are reviewed here. The carriage of these risk factors from South Asia to other countries has resulted in excess risk of oral cancer in these new settlements. There is also a high incidence of cardiovascular disease, hypertension and late onset diabetes among Indians living in the United Kingdom and there is new evidence to suggest that the combined roles of areca and ST may be contributory. Because of their enhanced financial situation, substance abuse may increase in their new country of domicile. The two products are psychologically addictive and a dependency syndrome related to their use among Asian immigrants to the United Kingdom has been described recently. PMID- 11900633 TI - Areca nut-abuse liability, dependence and public health. AB - To define a dependence syndrome may convey attention upon a public health problems hitherto relatively ignored. It may, however, stigmatize substance use, especially when western diagnostic criteria are applied to essentially culture bound substances. However, when the pattern of use of a substance results in significant personal harm, then whether dependence exists or not is less important than developing an appropriate response, if its use is associated with significant morbidity and mortality. Such is the case with areca nut, the fourth most commonly used drug in the world after tobacco, alcohol and caffeine. In this paper I will explore its use and properties with respect to its abuse and dependence liability and consider the associated implications for public health. PMID- 11900634 TI - Socio-economic aspects of areca nut use. AB - The socio-economic aspects of areca nut consumption have been overlooked. A narrative review was conducted to establish some of these features of areca nut consumption. Medline, Pubmed and the World Wide Web were searched using the terms: areca nut, betel nut, areca catechu and pan masala. Further analysis was conducted of datasets describing aspects of United Kingdom areca nut sales and consumption. South Asian economies at different stages of development have varying areca nut cultivation practices, employment opportunities and marketing strategies. Attempts at regulation of areca nut import and sales are described. Retail practice among the South Asian communities of the United Kingdom was found to reflect the diverse consumer practices current in their countries of origin. A study of areca nut consumption patterns and motivations among Bangladeshi women resident in East London identified differences between those chewing areca nut in paan with and without tobacco. Further research into the socio-economic aspects of areca nut consumption is needed which should be multidisciplinary in focus, of sound scientific quality and incorporating the opinions of consumers. PMID- 11900635 TI - Sociocultural aspects of areca nut use. AB - This paper aims to describe the sociocultural context of areca nut use through exploring people's norms and values and the meaning underlying a wide range of different practices. A historical review is followed by an examination of more recent evidence, including quantitative and qualitative research conducted in the United Kingdom. It is concluded that some reports involving the various uses of areca nut and the conclusions drawn are confusing and that future studies need to be more explicit. While a sound scientific background is required, important sociocultural and religious issues around areca nut use must also be understood if health promotion initiatives are to be considered. PMID- 11900637 TI - How safe is cloning? PMID- 11900636 TI - Areca nut and betel-quid chewing bibliography. PMID- 11900638 TI - Effect of culture media on in vitro development of cloned mouse embryos. AB - The effect of simple and sequential embryo culture media on the preimplantation development of mouse nuclear transfer (NT) embryos reconstructed with cumulus cell nuclei using a mechanical NT technique was studied. Blastocyst formation rate was evaluated using CZB medium and the sequential media G1/G2 and KSOM/G2. Arrested two- and three-cell NT embryos were Hoechst-stained to check for nuclear abnormalities. Nonmanipulated and sham-manipulated parthenogenetic embryos served as controls for, respectively, the medium and the handling technique. Rates of blastocyst formation for medium and handling control embryos were similar in CZB (58% and 61%), in G1/G2 (94% and 85%), and in KSOM/G2 (88% and 84%). Development of NT embryos was significantly impaired from the two-cell stage onwards, reaching the blastocyst stage at a rate of 5% in CZB, 14% in G1/G2, and 28% in KSOM/G2. Arrested two- and three-cell stage NT embryos showed a high rate of binucleation. These data demonstrate not only that NT embryos are more sensitive to in vitro culture conditions than parthenogenetic control embryos but also that selection of culture media can influence the preimplantation development of NT embryos. PMID- 11900639 TI - Regenerated bovine fetal fibroblasts support high blastocyst development following nuclear transfer. AB - Regenerated bovine fetal fibroblast cells were derived from a fetus cloned from an adult cow and passaged every 2-3 days. Serum starvation was performed by culturing cells in DMEM/F-12 supplemented with 0.5% FCS for 1-3 days. In vitro matured bovine oocytes were enucleated by removing the first polar body and a small portion of cytoplasm containing the metaphase II spindle. Cloned embryos were constructed by electrofusion of fetal fibroblast cells with enucleated bovine oocytes, electrically activated followed by 5 h culture in 10 microg/mL cycloheximide + 5 microg/mL cytochalasin B, and then cultured in a B2 + vero-cell co-culture system. A significantly higher proportion of fused embryos developed to blastocysts by day 7 when nuclei were exposed to oocyte cytoplasm prior to activation for 120 min (41.2%) compared to 0-30 min (28.2%, p < 0.01). Grade 1 blastocyst rates were 85.1% and 73.3%, respectively. The mean number of nuclei per grade 1 blastocyst was significantly greater for 120 min exposure (110.63 +/- 7.19) compared to 0-30 min exposure (98.67 +/- 7.94, p < 0.05). No significant differences were observed in both blastocyst development (37.4% and 30.6%) and mean number of nuclei per blastocyst (103.59 +/- 6.6 and 107.00 +/- 7.12) when serum starved or nonstarved donor cells were used for nuclear transfer (p > 0.05). Respectively, 38.7%, 29.4%, and 19.9% of the embryos reconstructed using donor cells at passage 5-10, 11-20 and 21-36 developed to the blastocyst stage. Of total blastocysts, the percentage judged to be grade 1 were 80.9%, 79.2%, and 54.1%, and mean number of nuclei per grade 1 blastocysts, were 113.18 +/- 9.06, 100.04 +/- 6.64, and 89.25 +/- 6.19, respectively. The proportion of blastocyst percentage of grade 1 blastocysts, and mean number of nuclei per grade 1 blastocyst decreased with increasing passage number of donor cells (p < 0.05). These data suggest that regenerated fetal fibroblast cells support high blastocyst development and embryo quality following nuclear transfer. Remodeling and reprogramming of the regenerated fetal fibroblast nuclei may be facilitated by the prolonged exposure of the nuclei to the enucleated oocyte cytoplasm prior to activation. Serum starvation of regenerated fetal cells is not beneficial for embryo development to blastocyst stage. Regenerated fetal fibroblast cells can be maintained up to at least passage 36 and still support development of nuclear transfer embryos to the blastocyst stage. PMID- 11900640 TI - Pluripotency of bovine embryonic cell line derived from precompacting embryos. AB - We report herein the establishment of three bovine pluripotent embryonic cell lines derived from 8-16-cell precompacting embryos. Two cell lines were cultured for 10 passages and underwent spontaneous differentiation. One cell line (Z2) has been cultured continuously for over 3 years and has remained undifferentiated. These cells express cell surface markers that have been used routinely to characterize embryonic stem (ES) and embryonic germ (EG) cells in other species such as stage-specific embryonic antigens SSEA-1, SSEA-3, and SSEA-4, and c-Kit receptor. In the absence of a feeder layer, these cells differentiated into a variety of cell types and formed embryoid bodies (EBs). When cultured for an extended period of time, EBs differentiated into derivatives of three EG layers - mesoderm, ectoderm, and endoderm - which were characterized by detection of specific cell surface markers. Our results indicate that the Z2 cell line is pluripotent and resembles an ES cell line. To our knowledge, this is the first bovine embryonic cell line that has remained pluripotent in culture for more than 150 passages. PMID- 11900641 TI - Nucleolar protein allocation and ultrastructure in bovine embryos produced by nuclear transfer from embryonic cells. AB - In the present study, immunofluorescence confocal laser scanning microscopy, autoradiography following (3)H-uridine incubation, and transmission electron microscopy were used to evaluate the nucleolar protein localization, transcriptional activity, and nucleolar ultrastructure during genomic reprogramming in bovine embryos reconstructed by nuclear transfer from in vitro produced bovine morulae to activated cytoplasts. During the first cell cycle (one cell embryos), no autoradiographic labelling was detected. Ultrastructurally, whorls consisting of densely packed fibrillar material were observed instead of nucleoli. During the second, third, and fourth cell cycle (two-, four-, and tentative eight-cell embryos), autoradiographically unlabelled nuclei contained vacuolated bodies consisting of densely packed fibrillar material. Also, during the fourth cell cycle, the first nucleoplasmic autoradiographic labelling was observed, but still without formation of fibrillo-granular nucleoli. During the fifth cell cycle (tentative 16-cell embryos), the nuclei displayed autoradiographic labelling over both nucleoplasm and presumptive nucleoli, and the formation of fibrillo-granular nucleoli was observed. In a certain proportion of blastomeres, however, abnormal patterns of nucleolar formation and apoptosis were noted. During the first two cell cycles, labelling of RNA polymerase I, fibrillarin, upstream binding factor (UBF), nucleolin (C23), and nucleophosmin (B23) was localized to nuclear entities. During the third cell cycle, labelling of topoisomerase I was observed in addition. During the fourth and fifth cell cycles, a substantial portion of the embryos presented blastomeres that lacked labelling of several of these nucleolar proteins. In conclusion, the nuclear transfer procedure was associated with remodelling of the nucleoli to an inactive form, followed by reformation of fibrillo-granular nucleoli during the fifth cell cycle. Moreover, a certain proportion of blastomeres failed to form functional nucleoli with respect to both ultrastructural organization and protein allocation. PMID- 11900642 TI - Placental anomalies in a viable cloned calf. AB - Placental anomalies are associated with a high mortality rate in mammalian cloning programs. In this report, we detail the very unusual occurrence of a grossly abnormal placenta that supported a viable cloned calf to term. The placenta was recovered intact 3 h following birth, and its weight was within normal limits (4.3 kg). The chorioallantois of the cloned transgenic female Holstein calf contained only 26 cotyledons. Twelve of these were enlarged and functional. Six were poorly developed, and eight were degenerating. The 12 functional cotyledons ranged in diameter from 8 to 20 cm. The nongravid horn had six rudimentary (<5 cm in diameter) cotyledons and eight cotyledons that remained as oval, mineralized plaques. Despite the reduction in number of placentomes, there was no adventitial placentation. Although this report documents observations from a single case, it does show that a morphologically deficient placenta was able to support development to term and resulted in a viable calf. PMID- 11900643 TI - Somatic cell cloning without micromanipulators. AB - Until now, micromanipulators have been regarded as indispensable for somatic cell nuclear transfer. This paper describes an improved zona-free nuclear transfer procedure with manual bisection of oocytes, selection of cytoplasts by Hoechst staining, and two-step fusion of somatic cells from primary granulosa cell cultures with two cytoplasts. Blastocyst rates in the three systems tested for zona-free embryo culture were 0%, 18%, and 36% for microdrops, well of the wells (WOW system), and microcapillaries (GO system), respectively. This simple, rapid, and inexpensive procedure may become a useful alternative to the existing techniques for somatic cell nuclear transfer for large-scale application of the technology. PMID- 11900644 TI - Expression and cellular localization of interleukin-8 mRNA and protein in the area of xenogenic bone implant. AB - OBJECTIVE: To observe the expression and cellular localization of interleukin-8 (IL-8) mRNA and protein in the area of xenogenic bone implant. METHODS: The bovine cancellous bone granules were implanted into the thigh muscles of mice. The samples were taken 4, 7, 14 and 21 days after implantation. IL-8mRNA and protein in the site of implant were assayed by in situ hybridization and immunocytochemical techniques. RESULTS: The expression of IL-8mRNA and protein were observed in all specimens 4, 7, 14 and 21 days after implantation. IL-8mRNA was expressed mainly by the neutrophils, monocytes, macrophages and fibroblasts at 7th day post-implantation. Some mesenchymal cells, multinucleated giant cells, vascular endothelial cells and smooth muscle cells also expressed IL-8mRNA in the area of xenogenic bone implant at 14th and 21st days. Immunocytochemical studies demonstrated the same results as that of in situ hybridization. CONCLUSIONS: Many different kinds of cells express IL-8mRNA and secret IL-8 in the area of xenogenic bone implant, suggesting that IL-8 may play an important role in local immunity of xenogenic bone graft. PMID- 11900645 TI - A primary study on TRISS in a Chinese hospital. AB - OBJECTIVE: To investigate a revision of the Trauma and Injury Severity Score (TRISS) weight coefficients in order to overcome the inference from foreign coefficients on Chinese trauma scoring. METHODS: The data of 1 297 Chinese trauma patients were studied for trauma scoring with the Revised Trauma Score-Injury Severity Score-TRISS (RTS-ISS-TRISS) system to get a serial of new weight coefficients through analyzing a multivariation logical regression between the outcome and the injury severity. RESULTS: ISS was higher but the Age Score and probability of survival (Ps) of the death group were lower than those of the survival group. New values of RTS-ISS-Age coefficients differed from the Major Trauma Outcome Study (MTOS) ones, through which the constant b(0) decreased its negative value, and ISS weight b(2) increased its negative value, but RTS weight b(1) and age weight b(3) changed with the trauma types. MTOS's values and new values of weight coefficients were used on 1297 patients for prognosis by calculating Ps. The accuracy of new values (90.13%) was a little higher than that of MTOS's (89.5%), with a promotion in specialization but a loss in sensitivity. CONCLUSIONS: A revision of TRISS's weight coefficients is either necessary or feasible. To achieve this purpose, a Chinese trauma database with large capacity is recommended. PMID- 11900646 TI - Preparation and assessment of heterotopic osteoinduction of beta-TCP/rhBMP-2 composite. AB - OBJECTIVE: To search for ideal bone graft substitute. METHODS: The beta TCP/rhBMP 2 composite was constructed by combining beta-Tricalcium phosphat (beta-TCP) that was prepared by the authors with recombinant human morphogenetic protein-2 (rhBMP 2) and was implanted into the muscle pouches in the thigh of mice. beta-TCP alone was implanted on the opposite side as controls. At intervals of 1,3,7,14 and 28 days after the implantation, the specimens were obtained, and histologic study and alkaline phosphatase assay (7,14,28 days) were performed. RESULTS: There was a large amount of cartilage and bone formation within the composite, increasing with time; whereas there was no new bone formation where beta-TCP alone was implanted. Besides, the levels of alkaline phosphatase in the beta-TCP/rhBMP-2 implants also were increasing with time and were higher than those in controls. CONCLUSIONS: The results indicate that beta-TCP/rhBMP-2 composite possesses heterotopic osteoinductive potential. PMID- 11900647 TI - The dynamic response of heart and its injury involving chest impact. AB - OBJECTIVE: The dynamic response of the heart during chest impact and the characteristics of heart injuries were investigated to further understand the mechanisms of heart impact injuries. METHODS: Eleven dogs and thirty-four rabbits were subjected to front thoracic impact with different impact velocities and compression response. The accelerated movement of thoracic wall during the impact period was monitored. The pathological examination of the injured heart was done and the dynamic responses and mechanisms of injuries were analyzed with mathematics models. RESULTS: The analysis of mathematics model and experimental results showed that the injury severity of heart was well correlated with the viscous criterion. The thoracic wall was involved in bi-directional movement of compression and expansion. The injured heart showed spotty or stripy hemorrhages in the ventricle endocardium. Light microscopic examination showed interstitial bleeding and rupture of the myocardial fibers in the contusion area. The biomechanical analysis indicated that there was a large deformation caused by the stress concentration on the lateral ventricle wall. CONCLUSIONS: There is a high speed and excessive deformation of the heart during the impact period, which might be the key mechanism of heart injury. The strong impact and press coming from both sternum and vertebral column and the rapid elevation of pressure in the ventricle are the main cause of deformation. PMID- 11900648 TI - Beneficial effects of fructose 1,6-diphosphate on hemorrhagic shock in rats. AB - OBJECTIVE: To investigate the effects of fructose 1,6-diphosphate (FDP) on experimental hemorrhagic shock in rats. METHODS: Sixty rats were randomly divided into three groups: the normal saline control group (group A), the 5% glucose solution control group (group B) and the 5% FDP solution treated group (group C). Shock models were made by bloodletting until the mean arterial pressure (MAP) reduced to 39.75 mmHg (1 mmHg=0.133 kPa) for 60 minutes, and then normal saline, 5% glucose and FDP were given to the rats, respectively. RESULTS: FDP could significantly increase MAP and the survival rate, elevate pH value, partial oxygen pressure (PaO(2)) and superoxide dismutase (SOD) activity, and decrease partial carbon dioxide pressure (PaCO(2)) and malondialdehyde (MDA) in arterial blood of the shocked animals. CONCLUSIONS: It suggests that FDP has a good protective effect on hemorrhagic shock by improving tissue metabolism and preventing acidosis and tissue injury caused by free radicals. PMID- 11900650 TI - Experimental study on early liver injury and expressions of TNF-alpha mRNA in burn rats with endotoxemia. AB - OBJECTIVE: To observe tissue distribution and cell localization of TNF-alpha mRNA and its protein and study their role in the pathogenesis of liver injury in burn rats. METHODS: An animal model of rats subjected to 20% TBSA III degree burns combined with intraperitoneal injection of lipopolysaccharide (LPS) was used for this experiment. The changes of hepatic morphology and functions and serum TNF alpha content and expression and localization of liver TNF-alpha and TNF-alpha mRNA were determined with light microscope (LM) and electron microscope (EM), quantitative analysis, immunohistochemistry (IHC) and in situ hybridization (ISH). RESULTS: It showed that there were sinusoid reaction, KCs activation and degeneration, necrosis of HCs, and platelets aggregation, fibrins deposition and PMNs attachment in sinusoid. The activity of ALT was obviously elevated and ALB content was slightly decreased. The serum content of TNF-alpha showed peak at 3 hours. TNF-alpha was mainly localized in sinusoid endothelial cells (SECs) and Kupffer cells (KCs), and TNF-alpha mRNA was mainly distributed in KCs, polymorphonuclears neutrophils (PMNs) and macrophages (MPs). CONCLUSIONS: It suggests that TNF-alpha mRNA and its protein expression and localization are coincident with the pathological changes of liver injury. TNF-alpha is one of the key cytokines in the pathogenesis of liver injury in burn rats with endotoxemia. PMID- 11900649 TI - Intra-axonal overloading of calcium ion in rat diffuse axonal injury and therapeutic effect of calcium antagonist. AB - OBJECTIVE: Exploring the intra-axonal overloading of calcium ion (Ca(2+)) in brain diffuse axonal injury (DAI) and the therapeutic effect of calcium antagonist(Nimotop) on DAI. METHODS: Fourteen SD rats were divided into injury group, treatment group and control group. The DAI model of rats was produced by using a head-instant-axial-rotation device. Tissues from the medulla oblongata of rats were taken 2-24 h post-injury and processed for electron microscopic observation by a cytochemical technique for calcium ion. RESULTS: In the injured rats there was evidence of local disruption of myelin sheath,lucent spaces between myelin sheath lamellae, separation of axolemma from the inner layer of myelin sheath, peripheral accumulation of organellae, intra-axonal formation of vacuoles and reduction of mitochondria. A large number of fine calcium deposits were seen on the affected myelin sheath. The severity of the myelin sheath lesion was related positively to the number of calcium deposits on it. In the later post injury period the coarse calcium particles appeared within the damaged axon. Neuronal somas and microvascular endotheliums showed a lot of vacuoles and some fine calcium deposits. Many microvilli formed on the luminal aspect of endothelium. In the treatment group myelin sheath tended to be injured locally, and axoplasmic mitochondria were nearly normal in number, structure, and distribution. Few calcium deposits were found in axons. Vacuolization was obviously reduced in neuronal soma and endothelium. CONCLUSIONS: In DAI there exists an intra-axonal overloading of calcium ion, which is a key factor to the occurrence and development of DAI. Early use of Nimotop can alleviate DAI. PMID- 11900652 TI - CT and MRI diagnosis of acute traumatic hepatic rupture: analysis of ten cases. AB - OBJECTIVE: In order to verify whether magnetic resonance imaging (MRI) is superior to computed tomography (CT) in the detection and characterization of intrahepatic hematoma in its acute stage, the MRI and CT features of acute traumatic hepatic rupture (ATHR) were retrospectively studied and compared. METHODS: In the 10 cases of ATHR admitted to our institute, 3 were examined with CT, 1 with MRI and 6 with both CT and MRI in the first 24 hours post injury and 9 cases out of the 10 were checked up with MRI in the first week after injury of surgery. The shape of the traumatic lesions, the damages of the intrahepatic vessels and the severity of hepatic rupture displayed with CT and MRI were compared. RESULTS: It was found that in the first 24 hours post injury, 66.6% of hepatic injuries were shown as hypointensity on T1-weighted images and low or high density on noncontrast CT. 100% of the lesions were identified as well marked hyperintensity on T2-weighted images. Damages of the hepatic and/or portal veins were observed in 7, 4 and 3 cases on T2- and T1-weighted images and noncontrast CT figures respectively. The severity of hepatic injuries were graded in 100%, 66.7% and 44.4%of cases with these 3 procedures respectively. CONCLUSIONS: On the basis of our findings, it is concluded that T2-weighted MRI is a more sensitive and reliable imaging modality in the detection and differentiation of the type and severity of acute hepatic rupture than T1 weighted imaging and noncontrast CT. PMID- 11900651 TI - Brain TXA(2) and PGI(2) levels in impact acceleration diffuse brain injury coupled with secondary insults. AB - OBJECTIVE: To study the changes of brain TXA(2) and PGI(2) levels in a new rodent model of impact acceleration diffuse brain injury with hypotention and hypoxia and the effect of diaspirin cross linked hemoglobin solution (DCLHb) on brain TXA(2) and PGI(2) levels. METHODS: Thirty-two male SD rats were randomized into sham, head injury alone, head injury with secondary insults and injury with insults followed by DCLHb administration groups. Animals were physiologically monitored throughout the experiment and the prostanoids were measured via radioimmunoassay (RIA). RESULTS: There were no changes in TXB(2) and 6-keto PGF1alpha (stable metabolites of TXA(2) and PGI(2)) levels in injury alone group while TXB(2) level in secondary insults group elevated significantly and both TXB(2) and 6-keto-PGF1alpha levels in injury with insults followed by DCLHb administration augmented significantly in comparison with the corresponding value of sham at 4 h postimpact. CONCLUSIONS: The only increase in TXA(2) level in secondary insults rats suggests that there may be both thrombotic episodes and vasoconstriction leading to focal increase in micro-circulatory resistance which contributes to a decreased focal cerebral blood flow (CBF). And it is hypothesed that DCLHb may exert its protective properties through increasing PGI(2) production in injured brain by affecting CBF and cerebral perfusion pressure (CPP). PMID- 11900653 TI - Significance of runaway calcium homeostasis in cultured cardiomyocytes in development of hypoxia, burnt serum-induced injury. AB - OBJECTIVE: This study aimed at understanding of the role of calcium homeostasis in cardiomyocytes from hypoxia, burnt serum-induced injury. METHODS: Alterations in cytosolic free calcium concentration (Ca(i)), calcium influx and viability of the cardiomyocytes in vitro after hypoxia, burnt serum stimulus were observed. RESULTS: Ca(i) increased markedly, in the meantime, the cellular transmembrane calcium influx increased and the viability of the cells decreased significantly following hypoxia, burnt serum-induced injury. CONCLUSIONS: In our study cytosolic calcium ion was transported abnormally in the cardiomyocytes after burn, to result in Ca(i) increase and runaway calcium homeostasis, thus the normal cellular function was disturbed. This may be one of the important factors in the development of burn-induced cardiac injury. PMID- 11900654 TI - Opioid receptors associated with cardiovascular depression following traumatic hemorrhagic shock in rats. AB - OBJECTIVE: To elucidate which one of &mgr;, delta and kappa opioid receptors is involved in the cardiovascular depression following traumatic hemorrhagic shock. METHODS: With traumatic hemorrhagic shock rat models, the changes of myocardial and brain &mgr;, delta and kappa opioid receptors and cardiovascular functions and their relationship with hemodynamic parameters were observed. The effects of delta and kappa opioid receptor antagonists on hemodynamic parameters of traumatic hemorrhagic shock rats were observed. RESULTS: Following traumatic hemorrhagic shock, the number of myocardial and brain delta and kappa opioid receptors significantly increased, their affinity did not alter, and the increased number of delta and kappa opioid receptors was significantly associated with the decreased hemodynamic parameters. However, &mgr; opioid receptor in heart and brain did not obviously change. delta opioid receptor antagonist ICI174,864 and kappa opioid receptor antagonist Nor-binaltorphimine (50 &mgr;g, Icv) could significantly reverse those decreased hemodynamic parameters. CONCLUSIONS: It suggests that opioid receptors, especially delta and kappa opioid receptors are closely related to the pathogenesis of traumatic hemorrhagic shock, and they play important roles in the depression of cardiovascular function following traumatic hemorrhagic shock. PMID- 11900655 TI - Osteogenic role played by fibroblasts in secondary healing of fracture. PMID- 11900656 TI - Hemoglobin-based blood substitute and its clinical application in trauma. AB - Blood transfusion is still an imperative and effective therapeutic remedy for many diseases, especially for trauma and shock. But the donation of blood by healthy citizens is seriously inadequate, and, more recently, blood transfusion is a matter of great concern because of viral infections such as hepatitis and HIV. In addition, blood transfusion has some other limitations including difficulty of storage and transportation, and needing cross-matching because of blood group antigens. Thus, the development of an oxygen carrier, which could serve as a safe and effective alternative to human blood or red blood cell (RBC), has gained great interest worldwidely. PMID- 11900657 TI - Using retrograde island flap with dorsal interosseous artery to repair soft tiss ue defect of hands. AB - Since the report of retrograde island flap with radial artery by Lu and Wang in 1982 and retrograde island flap with ulnar artery by Li in 1984, the flaps from forearm has been commonly used clinically in recent years because of its good textrue and satisfactory therapeutic results. However, whether the function of hand will be impaired by cutting one of the main arteries which supply the hand still remains controversial. There has been an acute ischemia of hand reported in foreign literature. Since 1985 we have studied 50 upper limbs from fresh cadavers by arteriograghy and vascular dye infusion in our hospital. Based on the results that the distal end of the dorsal interosseous artery of forearm can be anastomosed with a number of dorsal branches of volar interosseous artery at the back of the wrist, we have designed a retrograde island flap with these anastomotic branches as its pedicle to repair soft tissue defect of hands and applied in 71 cases. Seventy flaps survived, only one underwent necrosis because of vascular deformity. It is concluded that the flap has many advantages such as good texture, easiness of anatomical operation and high survival rate. What is more, the flap can be transplanted with nerve, muscle tendon and ulner artery without damage to the main artery of forearm. PMID- 11900658 TI - Clinical experience of the management for the most severely head-injured patients with GCS score of 3. AB - OBJECTIVE: To summarize the therapeutic experience of 24 patients of traumatic head injuries with GCS score of 3. METHODS: Twenty-four most severely head injured patients with GCS score of 3 who were admitted to our department from Jan 1995 to Mar 1998 were retrospectively analyzed. RESULTS: Twelve cases (50.0%) survived, of which 7 cases (29.2%) had good recovery or moderate disability and 5 cases with severe deficits (20.8%), and the other 12 died (50.0%) after therapy. CONCLUSIONS: The prognosis of most severely head-injured patients with GCS score of 3 could be improved by early intracranial hematoma removal with large decompressive craniotomies, early moderate hypothermia therapy, early assistant ventilation and effective prevention and treatment of complications. PMID- 11900659 TI - Free flap transfer bridged by antegrade and retrograde posterior tibial vessel flaps temporally borrowed from the healthy leg. AB - OBJECTIVE: To study the methods and techniques of free flap transfer bridged by posterior tibial vascular flap in treating large soft tissue defects in low limbs without usable recipient blood vessels. METHODS: Based on morphological observation and measurement of arterial pressure and blood flow, an antegrade and a retrograde vascular bridge flaps were designed using the healthy posterior tibial vessels to serve as vascular pedicles to carry two free flaps for transplantation. RESULTS: Eight cases of patient with one or two large soft tissue defects in the leg region were treated by the method. All the bridge flaps and free flaps survived well, and the defects were repaired completely. CONCLUSIONS: The results showed that it is an ideal method for using the posterior tibial vessels from the healthy limb to form vascular pedicles in repairing large soft tissue defects in patients without a usable recipient blood vessel. PMID- 11900660 TI - Osteogenic potential of rabbit dermal fibroblasts cultured in vitro: a scanning electron microscopic study. AB - OBJECTIVE: To investigate further into the osteogenic potential of rabbit dermal fibroblasts with scanning electron microscope. METHODS: Split-thickness rabbit skin was processed into small chips that were cultured in vitro and then subjected to scanning electron microscopic study. RESULTS: The fibroblasts swam out from the skin chips and they increased in number rapidly and became confluent. The cells exhibited squamous configuration, possessing arboreal bifurcation and forming multi-layer structure. The fibroblasts then excreted numerous minute granules, heaping up on and around the cells. Thenceforth emerged on the cell surface fine needle-like crystals, that agglomerated with the granules to form nodules. The fibroblasts orientated themselves in a radiating pattern around the large nodules. Neighboring nodules could be linked up into trabecular structures. CONCLUSIONS: Sequence of events of new bone formation by rabbit dermal fibroblasts cultured in vitro is fully depicted and confirmed. PMID- 11900661 TI - Penetrating wounds of the heart: an analysis of 61 cases. AB - OBJECTIVE: To present the managing experience of the patients with penetrating cardiac injuries for improving the treatment outcome. METHODS: The data of 61 cases with penetrating wounds of the heart were retrospectively studied, and the study covered a period of 11 years. RESULTS: In this series, stab wounds accounted for 61 cases (84%), among which, 9 cases with stable hemodynamics were managed conservatively, and the rest 52 underwent thoracotomy. The amount of preoperative infusion was less than 1000 ml in 65% of the latter cases. Only in two patients, preoperative pericardiocentesis was done, yielding false negative in one. Four cases sustaining cardiac arrest soon after arrival were subjected to emergency thoracotomy resulting in three survivals. Among 52 patients undergoing surgery, two deaths occurred after operation from associated abdominal injuries or failure of cerebral resuscitation. In the present series of 61 cases, the overall survival rate was 96.7%. CONCLUSIONS: Early establishment of diagnosis and prompt thoracotomy are the fundamental factors affecting the outcome of penetrating cardiac injuries. Preoperative massive transfusion and pericardiocentesis are not advocated. PMID- 11900662 TI - Effect of bactericidal/permeability-increasing protein on sepsis induced by intra abdominal infection in rats. AB - OBJECTIVE: To investigate the effect and mechanism of bactericidal/permeability increasing protein (BPI) on sepsis induced by intra-abdominal infection in rats. METHODS: Cecal ligation and puncture (CLP) was made on 20 rats with sepsis induced by intra-abdominal infection. BPI or equal volume of physiological saline (PS) was intra-abdominally given immediately and 12 h after CLP, respectively (2.5 mg/kg of BPI each time). Plasma endotoxin levels were determined with limulus amebocyte chromogenic assay. RESULTS: (1) The survival time in BPI group was significantly higher than that in PS group. (2) The values of the mean arterial pressure (MAP), the left ventricular systolic pressure (LVSP), the isovolumic ventricular pressure (IP), and the maximal change of left intraventricular pressure (+/-dp/dtmax) in BPI group, although decreasing, were markedly higher than those in PS group. (3) Plasma glutamic-pyruvic transaminase (GPT) and urea nitrogen levels in BPI group, though increasing, were obviously lower than those in PS group. (4) There was no significant change of plasma endotoxin levels in BPI group, while plasma endotoxin levels markedly increased in PS group. CONCLUSIONS: BPI has obvious protective effect on sepsis induced by intra-abdominal infection, which might be related to its neutralization of endotoxin. PMID- 11900663 TI - The relationship between porosity, collagen fiber orientation and strength of plated bone after rigid plate fixation and removal. AB - OBJECTIVE: To understand the relationship between porosity, collagen fiber orientation and strength of the plated bone after rigid plate fixation and removal. METHODS: Seventy-two New Zealand white rabbits were used in this experiment. Eight animals served as control and the other sixty-four were plated on their intact left tibiae with stainless steel (316L) 4-hole plates to induce early osteoporosis. The plates were removed 2 months after internal fixation in 40 plated animals, 8 of which were sacrificed immediately following plate removal and the other 32 were killed in successive groups with 8 in each group 1, 2, 3 and 4 months after plate removal. The remaining 24 plated animals were killed at 3, 4 and 6 months after plate fixation. After sacrifice, the samples of plated bone were prepared for light microscope, quantitative histological analysis, polarized light microscope and biomechanical test. RESULTS: The internal fixation with a rigid plate could induce the regional osteoporosis which manifested both bone loss and disorganized bone structure (loss of the orientation of the collagen fibers) leading to decreased strength of the plated bone. Although the regional osteoporosis could recover gradually after plate removal, the bone structure remained disorderly even when the bone mass returned to normal. Delayed restoration of bone structure was related to delayed restoration of bone strength. CONCLUSIONS: Besides the bone loss, the disorganized bone structure is the main cause of decrease of bone strength after rigid plate fixation and removal. PMID- 11900664 TI - Emergency treatment of craniocerebral firearm wounds. AB - OBJECTIVE: To improve the outcome of treatment in patients with craniocerebral firearm wound. METHODS: Prospectively and retrospectively reviewed a series of 93 patients presented to the Xi-Jing Hospital of Fourth Military Medical University with a diagnosis of craniocerebral firearm wound during a period of 27 years from July 1970 to July 1997. All the patients had acute craniocerebral firearm wound. Of these, it consisted of 81 males (87.1%) and 12 females (12.9%) ranging from 3 months to 58 years in age (median 24.6 years). The lesion included 16 tangential wounds, 58 tubular wounds and 19 through-and-through wounds. The cases were urgent and in serious and unstable condition. All the patients underwent surgical intervention and aggressive perioperative management in the neurosurgical intensive care, including resuscitative protocols. RESULTS: After emergency treatment and operation, 9 cases died (9.7%). Follow-up studies at three months postoperative showed that 56 cases (66.7%) had made good recovery. Rates of moderate disability, severe disability or vegetative state in this series were 19.0%, 10.7% and 3.6%, respectively. Long term follow-up studies (median 5.5 years) found that 42 (50.0%) were capable of resuming their occupation. CONCLUSIONS: Craniocerebral firearm wounds are often severe, needing urgent treatment for the patients. Timely, proper and thorough initial debridement are crucial for avoiding rapid neurological deterioration. PMID- 11900665 TI - Alterations of bcl-2, bcl-x and bax protein expressions in area CA-3 of rat hippocampus following fluid percussion brain injury. AB - OBJECTIVE: To investigate the alterations of bcl-2 gene family in the area of CA 3 in rats and the molecular mechanism of neuronal apoptosis following traumatic brain injury. METHODS: To investigate the alterations of bcl-2 gene family in the area of CA-3 in rats and the molecular mechanism of neuronal apoptosis following traumatic brain injury. RESULTS: The immunoreactivity of bcl-2 and bcl-x proteins decreased in the hippocampus ipsilateral impact site at 6 hours after injury, and this was the main cause of down-regulation of the value of (bcl-2+bcl-x)/ bax. During the period of 1-3 days after injury, bax protein expression increased significantly, while bcl-2 and bcl-x protein expressions decreased relatively slowly. The decreased value of (bcl-2+bcl-x)/ bax was mainly due to the bax up regulation. CONCLUSIONS: The bcl-2 gene family is involved in neuronal apoptosis after traumatic brain injury, and the protein-expression alterations of the bcl-2 gene family members lead to apoptosis of the neuronal cells. PMID- 11900666 TI - Ultrastructural changes of TMJ articular cartilage and synovial membrane following occlusal trauma in rabbit. AB - OBJECTIVE: To study the effects of occlusal trauma on the ultra-structure of synovial membrane and articular cartilage in rabbit's temporomandibular joints (TMJ). METHODS: TMJs from six rabbits with occlusal trauma and three control rabbits were studied by transmission electron microscopy. RESULTS: Degenerative changes in synovial membrane and articular cartilage of TMJ were induced following occlusal trauma. The structure of the articular surface was damaged, and chondrocytes in cartilage showed signs of degeneration. The synovial lining cells contained dense accumulations of vimentin intermediate filaments (IFs), which were especially prevalent in the cellular processes as well as paranuclearly. Microvilli on the synovial cell membrane were commonly seen. The "vermiform bodies" in the deeper interstitium of the synovial tissue were also found. Our findings of the punctate adherens between synovial lining cells were described in detail. CONCLUSIONS: The occlusal trauma is really a factor inducing degenerative changes of the TMJ. PMID- 11900667 TI - Schwann cells and fibronectin treating lesioned spinal cord of adult rats. AB - OBJECTIVE: To determine whether Schwann cells (SCs) and fibronectin (FN) support the growth of damaged axons and their conductive function. METHODS: Thirty adult Sprague-Dawley (SD) rats were randomized into 3 groups (n=10), and the following grafts: both SCs and FN, FN, and basal medium were implanted respectively into the lumbar of spinal cords hemisected at vertebra T(12). RESULTS: At 6 weeks postoperation, the latencies of spinal cord evoked potential (SCEP) P(1) wave in the three groups were 1.57 ms, 1.84 ms, and 2.03 ms, respectively. The differences between SCs/FN group and the other two groups were statistically significant. The number of regenerated axons in SCs/FN group was significantly greater than that in FN group. The number of survival neurons in L(4) and L(5) left dorsal root ganglia (DRG) in SCs/FN group was significantly greater than that in the rest two groups respectively. The latencies of P(1) wave were significantly correlated with axon counts in both SCs/FN and FN groups. CONCLUSIONS: SCs/FN grafted to the hemisected lumbar of spinal cords can produce robust axon regeneration and promote partial repair of their conduction. Surface recording of SCEP technique has been proved to be a reliable and less-traumatic method for assessment of recovery of afferent conduction in hemisected spinal cord. PMID- 11900668 TI - Analysis of the unexpected damage in children Wilm's tumor. AB - OBJECTIVE: To explore the relationship between unexpected damage and clinical character, biological activities and molecular biology. METHODS: We treated 76 cases of children wilm's tumor from 1995 to 1998. Among them, 9 cases (11.8%) underwent unexpected damages. We studied and analyzed the clinical characters, clinical pathology, morphology and molecular biology in the 9 cases. RESULTS: Wilm's tumor in late stage, blastoma cell type and bad differentiation was likely to be damaged unexpectedly, and the outcome was poor. CONCLUSIONS: Mutation of gene is related to the biological activities. It may be a potential and important factor to cause unexpected damage. PMID- 11900669 TI - Balloon occlusion test and therapeutic occlusion on traumatic carotid cavernous fistulas. AB - OBJECTIVE: To evaluate the safety of the balloon occlusion test(BOT) and therapeutic occlusion of the internal carotid artery(ICA). METHODS: The data of 43 patients hospitalized consecutively with traumatic intractable carotid cavernous fistulas (TICCF) were analyzed. Therapeutic occlusion of ICA was performed on 39 cases and BOT was only performed on the remaining 4 cases. Our assessment consisted of: (1) angiographic evaluation of collateral circulation with or without BOT of ICA, and (2) evaluation of clinical tolerance to therapeutic occlusion of ICA with hypotensive challenge for 30 minutes. Complications of BOT and therapeutic occlusion of ICA were also analyzed retrospectively. RESULTS: Complications related to BOT occurred in 1 case (2.3%) without causing permanent deficits. Complications related to therapeutic occlusion of ICA occurred in 4 cases (10%), including 1 technical (2.5%), 2 temporary (5%) and 1 permanent (2.5%) deficit. There was no fistula recurrence or mortality. CONCLUSIONS: BOT of ICA is safe and economical. The reliability of the results is almost the same compared with that of other more complicated methods of assessing therapeutic occlusion of ICA. And it is easy to treat TICCF with therapeutic occlusion of ICA. PMID- 11900670 TI - Biomechanical experiment and clinical report of modified patellectomy for polar fracture of the patella. AB - OBJECTIVE: To study the influence of polar patel-lectomy and the site of reanchored the patellar tendon on femoral quadriceps extensing force. METHODS: The experiment was designed by using 5 lower limbs of cadavers for extracorporal biodynamic test. The maximal fixing force of the double stainless steel wire (0.4 mm in diameter) reanchoring the patellar ligament to the patellar cortical edge was measured. Clinically 21 patients with polar fractures were treated by removing the small fragments and reanchoring the patellar ligament to the cortical edge of the patella using the wires. RESULTS: Resection of the lower 1/4 part of the patella and the patellar ligament reanchored to cortical edge of the patella would not increase femoral quadriceps contracting force arm. But if the patellar tendon reanchored near the joint surface, much more extending force would be needed than the former (P<0.001). The maximal fixing force of the wires is 67.4 kg (60.9-72.5 kg). All the patients using this modified procedure gained satisfactory functional recovery. CONCLUSIONS: The modified partial patellecomy is a simple and effective procedure for choice. PMID- 11900671 TI - Traumatic lacunar infarction in basal ganglion in children. AB - Traumatic lacunar infarction in basal ganglion in children under 10 year-old is more liable to occur. Before CT was available, the nature and localization of the lesion in this region had been difficult. It is sometimes difficult to define a correct treatment. Here we report 32 cases of this disease we have treated in recent years. PMID- 11900672 TI - Anterior decompression of late thoracolumbar fracture dislocation with paraplegia. AB - The operative methods for treatment of thoracolumbar fracture-dislocations have been improved in recent years. Anterior or anterior-lateral decompression for thoracolumbar fracture-dislocations with spinal cord injury is considered as a practical and successful method by many researchers. Since 1994 using trans pedicular partial vertebrectomy, we have treated 43 cases of late thoracolumbar fracture-dislocations with paraplegia successfully. PMID- 11900673 TI - Diagnostic strategy in patients with extraesophageal GERD. PMID- 11900676 TI - Nutritional management of acute pancreatitis. AB - Most patients with acute pancreatitis have mild to moderate disease and require no specialized nutritional support. Twenty percent to 30% have severe cases, resulting in a catabolic hypermetabolic state, and these patients may require early aggressive nutritional support. Traditionally, this support has been in the form of total parenteral nutrition. However, recent data suggest that enteral nutrition infused into the jejunum is feasible, well tolerated, associated with fewer complications, and significantly less expensive than parenteral nutrition. The pathophysiology of gut function in acute pancreatitis and the rationale and evidence for parenteral and enteral nutritional support are reviewed herein. An algorithm on the nutritional management of acute pancreatitis is suggested. PMID- 11900675 TI - Predicting and preventing post-ERCP pancreatitis. AB - Pancreatitis is rightly the most feared complication of endoscopic retrograde cholangiopancreatography (ERCP). Ten percent to 15% of cases of post-ERCP pancreatitis (PEP) are severe by clinical and radiologic criteria. Such cases carry significant morbidity and mortality and are responsible for the vast majority of ERCP-related deaths. The prediction and prevention of PEP have been of great interest to endoscopists since the introduction of ERCP 30 years ago. Prediction and diagnosis of PEP have become more accurate with the widespread availability of serum amylase estimation. A variety of cytokines (eg, interleukin -1, IL-6, and IL-8) and acute phase reactants (eg, C-reactive protein) are also elevated in the serum in acute pancreatitis, and these form the basis of evolving tests for PEP. Urine testing (for amylase) in acute pancreatitis is obsolete, but it may soon undergo a revival in the form of a rapid (3-minute) dipstick test for trypsinogen-2, a sensitive and specific test for this disease. The prevention of PEP takes multiple forms. The following steps are recommended for clinicians: 1) avoid ERCP when other, less invasive or noninvasive imaging tests can do the job (eg, CT or magnetic resonance imaging); 2) avoid high-risk (of PEP) procedures, such as needle-knife papillotomy, balloon dilation of the biliary sphincter, and pancreatic sphincterotomy, and take steps to reduce risk when these procedures are unavoidable; 3) ensure that those who perform ERCP have adequate training and experience; and 4) consider pharmacologic intervention. Despite a depressing catalog of drug interventions that have failed over the years (eg, antihistamines, anticholinergics, and corticosteroids), three agents have recently shown promise: somatostatin; its octapeptide analogue, octreotide; and gabexate mesylate, a protease inhibitor. PMID- 11900677 TI - Evidence-based approach to idiopathic pancreatitis. AB - The causes of recurrent acute pancreatitis, including the newly recognized genetic causes, are reviewed. The pitfalls of overcalling the diagnosis of pancreatitis in patients with abdominal pain or other symptoms and the role of a careful history are emphasized. The presence of undetectable microlithiasis in patients with unexplained pancreatitis is discussed. The popular notion of pancreas divisum and sphincter of Oddi dysfunction as causes of pancreatitis is challenged, as is the role of endotherapy in these conditions. PMID- 11900678 TI - Differentiating pancreatic cancer from pseudotumorous chronic pancreatitis. AB - The differentiation between pancreatic carcinoma and pseudotumorous pancreatitis continues to be a challenge. Several diagnostic imaging and endoscopic modalities can assist in making the differentiation, but the accuracy of each method varies. Radiologic imaging techniques include transabdominal ultrasound, computed tomography, magnetic resonance imaging, magnetic resonance cholangiopancreatography, and positron emission tomography. Endoscopic techniques include endoscopic ultrasonography, intraductal ultrasonography, and endoscopic retrograde cholangiopancreatography with brush cytology of pancreatobiliary strictures, endoscopic forceps biopsy, and analysis of pancreatic juices for malignant cells. Tumor markers appear to be promising, but further studies are needed to define the role of these markers. PMID- 11900679 TI - Advances in imaging for pancreatic disease. AB - Pancreatic imaging is an essential tool in the early diagnosis and staging of pancreatic disease. This review analyzes the most recent advances in pancreatic imaging. The specific modalities discussed include helical computed tomography (HCT) and multislice CT (MSCT), CT angiography, magnetic resonance imaging (MRI), magnetic resonance cholangiopancreatography (MRCP), and positron emission tomography (PET). At present, MSCT is generally viewed as the most efficient modality for initial detection and staging of pancreatic carcinoma, with an accuracy rate of about 95% to 97% for initial detection and virtually 100% for staging. CT is also the initial imaging modality used in evaluation of acute pancreatitis. However, recently, MRI has been viewed increasingly as a more precise diagnostic tool in this subgroup of patients. MRCP has been accepted as the primary imaging technique in the diagnosis of chronic pancreatitis. PET imaging, on the other hand, has an increasing role in the staging of pancreatic carcinoma, for which it may be the modality of choice in detection of extrapancreatic metastasis. PMID- 11900681 TI - Sphincter of Oddi (pancreatic) hypertension and recurrent pancreatitis. AB - Major papilla pancreatic sphincter dysfunction, a variant of sphincter of Oddi dysfunction, causes pancreatitis and pancreatic-type pain. The gold standard for diagnosis is sphincter of Oddi manometry, most commonly performed at endoscopic retrograde cholangiopancreatography (ERCP). Noninvasive testing, such as secretin stimulated transabdominal or endoscopic ultrasound assessment of pancreatic duct diameter, is less reliable and has relatively low sensitivity. Two thirds of patients with biliary sphincter of Oddi dysfunction have elevated pancreatic basal sphincter pressure. To maximize the diagnostic yield of sphincter of Oddi dysfunction, both the biliary and pancreatic sphincter pressures should be measured. Patients with sphincter of Oddi dysfunction may respond to biliary sphincterotomy alone, but evaluation of their pancreatic sphincter is warranted if symptoms persist after biliary therapy alone. Whether both biliary and pancreatic sphincters should be treated at the first ERCP session is controversial. Biliary and pancreatic endoscopic sphincterotomies are associated with two- to fourfold increased incidence of pancreatitis following the procedure in patients with pancreatic sphincter hypertension. Prophylactic pancreatic duct stenting reduces the frequency and severity of complications by greater than 50%. PMID- 11900682 TI - The role of magnetic resonance cholangiopancreatography in patients with suspected biliary obstruction. AB - Endoscopic retrograde cholangiopancreatography (ERCP) is an accepted and accurate procedure that combines the advantage of diagnosis of biliary obstruction with possible therapeutic endobiliary intervention. However, it is an operator dependent and invasive procedure that is associated with complications and limitations. Magnetic resonance cholangiopancreatography (MRCP) is a unique noninvasive technique for the diagnosis of biliary obstruction. It is well suited to provide the information required to plan the optimal therapeutic approach for these patients. MRCP has the potential to replace or at least precede ERCP as the first-line imaging effort in the evaluation of suspected biliary obstruction. Significant advantages and some notable limitations inherent to the modality dictate its judicious use in appropriate circumstances. The present article reviews the utility of MRCP in evaluation of biliary obstruction, with brief reference to its principles and techniques. PMID- 11900683 TI - The role of microorganisms in biliary tract disease. AB - The biliary tract is normally sterile, but bile-tolerant bacteria are frequently isolated from patients with cholecystitis. Since the identification of about 25 Helicobacter species, some of which may grow in bile, studies have addressed the role of these organisms in primary biliary cirrhosis, primary sclerosing cholangitis, and cholelithiasis. Most of these bacteria show the presence of Helicobacter DNA or antigens in the bile tract and in liver samples. Altogether, data from studies on biliary and hepatic diseases, as well as pancreatic disorders, suggest that bile-tolerant Helicobacter species may induce a chronic infection with possible malignant transformation. PMID- 11900685 TI - [Thoracic sympathectomy by videothoracoscopy: an update]. PMID- 11900684 TI - Sphincter of Oddi dysfunction: is there a role for medical therapy? AB - If the mechanism of pain in patients with sphincter of Oddi (SO) dysfunction is functional obstruction of the biliary tract, and at least in some patients it results from sphincter smooth muscle hypertrophy, then smooth muscle relaxants should have a theoretic role in the management of these patients. Calcium channel antagonists and other smooth muscle relaxants have been shown in "acute" manometric studies to alter SO motility. However, the effect of these agents on vascular smooth muscle remains a concern and often limits their use. At present the role of medical therapy is somewhat unclear because few well-conducted studies have used manometric criteria for the diagnosis of SO dysfunction and the selection of patients for therapy. The main drawback is that no drugs appear to be specific for the SO, long acting, and free of side effects. PMID- 11900686 TI - [Construct validity of the Information and Decisions in Asthma questionnaire]. AB - OBJECTIVE: To establish the construct validity of a new scale based on the Information and Decisions in Asthma (IDEA) questionnaire, which was designed to detect an asthmatic's desire to receive information, active seeking out of information and ability to make decisions about his or her disease. METHODOLOGY: The IDEA questionnaire containing 31 items in three subscales: desire for information (DI), seeking out information (SI) and decision making (DM) was answered by 120 asthmatic adults (54 men and 66 women; 86 intrinsic and 44 extrinsic) in stable condition and with varying levels of severity of disease. After interviews to take down medical histories (age of onset, years of evolution, visits to the emergency room within the past year), educational level and economic status, all patients completed a battery of tests assessing the following: quality of life (QL), alexithymia (TAS-20), health opinions (HOS), state-trait anxiety (STAI-E/R), asthma symptoms control (ASC), health locus control (HLC), somatosensory amplification (SAS) and personality (NEO-PI). RESULTS: Although most patients expressed interest in obtaining information, their level of active seeking and decision making were much lower. DI and SI were associated with higher educational levels and economic status, whereas DM correlated with the presence of atopy, age (young patients), sex (women) and few visits to the emergency room. Multiple regression analysis of psychological variables showed that DI and SI were accounted for mainly by the absence of alexithymia, whereas DM depended on an individual's behavioral involvement with his or her disease. CONCLUSION: The global analysis of these results indicate that the IDEA questionnaire has satisfactory construct validity, given that its dimensions are associated with sociodemographic and psychological characteristics that can be expected given its conceptual content. PMID- 11900687 TI - [Dyspnea in COPD: relation to the MRC scale with dyspnea induced by walking and cardiopulmonary stress testing]. AB - OBJECTIVE: Exercise-related dyspnea is the main symptom of chronic obstructive pulmonary disease (COPD), yet its relation to lung function deterioration is weak. The aim of this study was to evaluate the relation between the patients' usual level of dyspnea and dyspnea caused by a maximum cardiopulmonary stress test or a 6-minute walking test. METHODOLOGY: Thirty-six consecutive patients with stable COPD (age 66 7 years post-bronchodilator FEV1 47 14% of predicted) were studied. In addition to full baseline function testing, all patients underwent stress testing on a cycle ergometer and a 6-minute walking test in a corridor 50 m long. Exercise-induced dyspnea was assessed by the patient on a Borg scale before beginning and after completing each test. Chronic dyspnea during activities of daily living was quantified on the Medical Research Council (MRC) scale. RESULTS: The MRC value was only weakly related to percent of predicted FEV1 (r = 0.34, p = 0.04). Parameters obtained during exercise tests that were associated with the MRC were SaO2 at the end of the 6-minute walking test (r = 0.49, p = 0.004) and change in dyspnea on the Borg scale during the 6 minute walking test (deltaBorg-6mWT, r = 0.54, p = 0.0008) and during the stress test (r = 0.35, p = 0.04). Multiple regression analysis, with the MRC result as the dependent variable, showed that deltaBorg-6mWT and SaO2 at the end of the walking test explained 29% of the variance. CONCLUSION: Severity of chronic dyspnea in COPD patients assessed on the MRC scale is more related to dyspnea triggered by the walking test than with dyspnea induced by cycle ergometer stress testing. PMID- 11900688 TI - [Factors associated with hospital mortality in patients admitted to the intensive care unit in Colombia]. AB - OBJECTIVES: To describe the demographic features, reasons for hospital admission and factors associated with hospital mortality in patients admitted to intensive care in Colombia. METHOD: A cohort study of patients admitted to intensive care units (ICUs). Of 89 ICUs identified in Colombia, 20 in ten cities were invited to gather information on 200 consecutive patients admitted to each ICU. RESULTS: Three thousand sixty-six patient cases were available for analysis. The mean age was 53 years and 43% were women (men vs. women, p < 0.001). The most frequent cause of admission was medical (63.9%), acute myocardial infarction patients (7.1%) comprising the largest group. Severity of disease measured as APACHE II and III was a mean 14.0 (SD 6.9) and 48.3 (SD 23.5), respectively. Multivariate analysis, independent of adjustment for severity (APACHE II or III), showed that the factors associated with hospital death were the need for mechanical ventilation, pupillary response, transfer from a medical ward, and management by the ICU team prior to admission (p < 0.01). CONCLUSION: The most common reason for admission to an ICU in Colombia was myocardial infarction. Besides severity of disease, other variables related to medical care in Colombia are associated with hospital mortality, such as invasive ventilation. Although these variables may be artifacts related to disease severity, they are more likely to be related to quality of care. PMID- 11900690 TI - [Muscle function in other obstructive pulmonary diseases, in malnutrition and in sepsis]. PMID- 11900689 TI - [Influence of computed tomography of the abdomen for staging lung cancer]. AB - OBJECTIVES: To analyze the influence of routine imaging of the upper abdomen by conventional computed tomography (CT) to stage bronchopulmonary carcinoma and to detect liver or adrenal metastasis. A second objective was to describe the characteristics of a large group of patients in our practice. MATERIAL AND METHODS: Retrospective study of 387 patients (367 men and 20 women; mean age [+/ SD] 62.3 +/- 10.4 years, range 34-90 years) who had received a diagnosis of lung cancer (203 epidermoid carcinoma, 75 adenocarcinoma, 15 non-small cell carcinoma, 68 small cell carcinoma and 25 mixed tumors). CT images were obtained of the chest and upper abdomen with intravenous contrast except in patients with a history of allergy or renal insufficiency. The characteristics associated with abdominal CT images aiding or confusing diagnosis were analyzed by Spearman coefficient. Differences related to sex or histology were studied using a Mann Whitney U-test and Kruskal-Wallis test. RESULTS: The upper abdominal CT changed the staging of 27 patients (7%): non-small cell carcinoma 5% (16/319) and small cell carcinoma 16.2% (11/68). Twelve patients (3.1%) showed evidence of unconfirmed adrenal or hepatic metastasis. Change of staging after CT was associated with a high creatinine concentration in blood (p = 0.032), whereas confusion of diagnosis after CT was more common for women (p = 0.002) and patients for whom the diagnosis was established by cytology of sputum or bronchial aspirate (p = 0.019). Differences between men and women were found for from pathology (p = 0.027), confusion after CT (p = 0.002), hemoglobin (p = 0.011), hematocrit (p = 0.019) and smoking (p = 0.000). CONCLUSION: Given the considerable limitations of CT imaging of the upper abdomen, new technologies should be developed to facilitate a more rational approach to the problem. PMID- 11900691 TI - [Function of respiratory muscles in malnutrition and in the critically ill patient]. PMID- 11900692 TI - [Passive smoking in adults]. PMID- 11900694 TI - [Bronchocentric granulomatosis and pulmonary hydatidosis]. AB - Bronchocentric granulomatosis is a non-specific necrotizing granulomatous reaction in the bronchi and bronchioles. In asthmatic patients, bronchocentric granulomatosis is considered a hypersensitivity reaction to intrabronchial fungi. However, in non-asthmatic patients the possible etiology has not been identified. We report the case of a 46-year-old woman with bronchocentric granulomatosis associated with pulmonary hydatidosis. The likely immuno-pathogenic mechanisms leading to bronchial lesions are briefly discussed. PMID- 11900693 TI - [Pure red cell aplasia in a lung transplant patient]. AB - Aplastic anemia secondary to infection by parvovirus B19 is normally an extremely rare problem in patients with no prior history. However, the presence of certain risks, such as receiving chronic immunosuppressant therapy, may facilitate its appearance. Very few cases have been published concerning red cell aplasia due to parvovirus B19 infection in patients receiving a transplanted lung. We report the case of a 24-year-old woman with cystic fibrosis who had received a double lung transplant. The patient developed red cell aplasia secondary to parvovirus B19 infection; severe anemia requiring multiple transfusions. Five days of intravenous immunoglobulin therapy resolved the anemia. We discuss the difficulty of serological diagnosis in such cases, the importance of using techniques that identify the virus and taking measures that may prevent recurrence. PMID- 11900695 TI - [Pulmonary disease associated with fluoxetine administration]. PMID- 11900696 TI - [Actinomycosis as the cause of pleural effusion]. PMID- 11900697 TI - [Costal hemangioma: a rare diagnosis]. PMID- 11900698 TI - [Ovarian hyperstimulation syndrome as the cause of pleural effusion: a case report]. PMID- 11900700 TI - [Antithrombotic treatment in hypertensive patients with chronic atrial fibrillation. CARDIOTENS 99 study]. AB - BACKGROUND: Our main goals were to know the actual degree of oral anticoagulation and antiaggregation in hypertensive patients with atrial fibrillation in the daily clinical practice in Spain and to analyze any differences between primary care physicians and cardiologists. PATIENTS AND METHOD: 32,051 outpatients attended the same day by 1,159 physicians (21% cardiologists) were prospectively included in a database taking into account a history of hypertension and atrial fibrillation, demographic data and ongoing treatments. RESULTS: Hypertension was detected in 10,555 patients and 999 of them had both hypertension and atrial fibrillation (9.46%: 435 males [44%] and 564 females [56%]). 53% patients were attended by primary care physicians and the rest by cardiologists. 33% of hypertensive patients with atrial fibrillation were on oral anticoagulation: 41% of them attended by cardiologists and 26% by primary care physicians (p < 0.05). These differences persisted when the patients were compared on the basis of their age. 39% of hypertensive patients were on oral antiaggregation treatment, without differences in both groups except for those aged less than 65 years who were found to receive more antiaggregation in primary care (36% vs 24%; p < 0.05). CONCLUSIONS: The prevalence of atrial fibrillation in hypertensive patients is about 10%; there is a suboptimal degree of utilization of oral anticoagulation, which is more evident in patients attended by primary care physicians; elderly patients (> 80 years-old) were found to receive less anticoagulants and more antiaggregants both in primary health-care and cardiology health-care. PMID- 11900699 TI - [Effect of captopril on left ventricular diastolic dysfunction in young insulin dependent diabetic patients with microalbuminuria]. AB - BACKGROUND: Diabetic cardiomyopathy (DMC) is a complication of diabetes mellitus (DM)that is more frequently observed in those patients with microalbuminuria. Left ventricular diastolic dysfunction (LVDD) in patients with diabetes, in absence of another etiology that justifies it, is an early marker of DMC. We carried out a prospective study on young diabetic type 1 patients with microalbuminuria, aimed at knowing the effect of captopril on LVDD. PATIENTS AND METHOD: We included 30 patients (18 males and 12 females) diagnosed with type 1 DM,aged 40 years old, who had been recently found to have microalbuminuria and thus they were candidates to receive captopril. We excluded patients having factors different from DM that could modify the diastolic function. All patients underwent a complete biochemical and echocardiographic study before starting the treatment with captopril and six months later. A diagnosis of LVDD was made when at least one of the following parameters was present in the echocardiographic study: isovolumetric relaxing time (IRT) >100 ms, deceleration time (DT) > 220 ms or early filling rate peak/late filling rate peak ratio (E/A) < 1. According to the results of the second echocardiogram, patients were classified in two groups: improved group (when there was at least a 10% improvement of initial LVDD altered parameters) and non-improved group. A control group of 28 type 1 diabetic patients without microalbuminuria who were not given captopril was included (group C). RESULTS: The initial echocardiographic study yielded 11 patients having a normal diastolic function (group FDN) and 19 patients having LVDD (group FDA). After 6 moths of captopril treatment, an improvement of the ratio E/A was observed in the group FDN: from 1.58 (0.36)in the beginning to 1,68 (0.29) six moths later (p < 0.05),and in the group FDA: from 1.09 (0.24) to 1.24 (0.28) (p <0.05). In the group FDA, an improvement of IRT was found: from 110 (16) ms to 99.9 (9.6) ms (p < 0.01). Moreover, in the group FDA, LVDD improved after sixth months in 15 (78.9%) patients but not in 4 (21,6%). This LVDD improvement was associated with a decrease of the diastolic blood pressure (DBP) and the systolic blood pressure (SBP) at the end of the study. A logistic regression analysis showed an independent association between the reduction of the mean SBP and the improvement of LVDD. CONCLUSION: Our results suggest that captopril can improve LVDD in young patients with type 1 diabetes and microalbuminuria, possibly due to a decrease of blood pressure. PMID- 11900702 TI - [Statins: benefit or risk?]. PMID- 11900701 TI - [Influence of asthma inhalers on a breath alcohol test]. AB - BACKGROUND: People and authorities are worried about interferences in breath alcohol measurements. The influence of different inhalers on breath alcohol tests is analysed in this paper. PATIENTS AND METHOD: We included 60 patients treated with different inhalers. They underwent a breath alcohol test before treatment and 1, 2, 3, 5 and 10 min after it. RESULTS: Every medication led to positive readings within the first minutes of its administration. However, all values reached zero at 10 min. Patients administered higher doses and those who did not use a spacer device were found to have higher reading values. CONCLUSIONS: Asthma inhalers, including those without alcohol contents, lead to positive readings of breath alcohol measuring devices within the first minutes. However, these interferences are no longer detected at 10 min. PMID- 11900703 TI - [Ethics and health management]. PMID- 11900704 TI - [New therapeutic strategies in liver fibrosis: pathogenic basis]. PMID- 11900705 TI - [Angiotensin II receptor antagonists in cardiac failure treatment]. PMID- 11900706 TI - [Visual hallucinations in the elderly]. PMID- 11900707 TI - [Folic acid supplementation in the prevention of neural tube defects]. PMID- 11900709 TI - [Primary prevention of neural tube defects: folic acid or levofoline acid?]. PMID- 11900711 TI - A symposium: National Cholesterol Education Program Adult Treatment Panel III: Impact and implementation of the new guidelines. Introduction. PMID- 11900713 TI - Strategies to improve Adult Treatment Panel III guideline adherence and patient compliance. AB - The National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III report outlines the management of hypercholesterolemia through guidelines. These guidelines call for more aggressive diagnosis and treatment of hypercholesterolemia, which will substantially increase the number of individuals in the United States considered to be at risk for heart disease and will expand the number who will receive dietary and drug treatment. The new features of ATP III add complexity to the guidelines, which will impact adherence as well as add challenges to the management of hypercholesterolemia. Following key recommendations and incorporating essential elements of adherence can improve implementation of the NCEP ATP III guidelines. The use of global risk scoring aids, including the Palm Pilot cholesterol risk calculators, can improve guideline adherence and provide education and motivation to patients to maintain compliance. PMID- 11900712 TI - Adult Treatment Panel II versus Adult Treatment Panel III: what has changed and why? AB - The Third Report of the Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel [ATP] III) differs in several ways from the ATP II guidelines. Several principal advances include (1) new risk levels for major lipid measures, (2) increased emphasis on primary prevention, (3) inclusion of high-risk groups in secondary prevention, (4) broader lifestyle program, and (5) increased focus on implementation and adherence. The purpose of this article is to discuss the major changes in ATP III and to highlight the benefits of the new guidelines in the management of hypercholesterolemia in adults. PMID- 11900715 TI - Cardiovascular risk management in clinical practice: the Midwest Heart Specialists experience. AB - Evaluating cardiovascular risk and managing hypercholesterolemia are essential components in the identification and treatment of coronary artery disease. A collaborative care-based lipid clinic is a beneficial part of an integrated prevention program and can result in improved lipid management. Advances in technology now enable use of the electronic medical record in combination with a virtual lipid clinic to promote superior cholesterol management. PMID- 11900717 TI - Noninvasive surrogate markers of atherosclerosis. AB - Noninvasive imaging techniques offer a unique opportunity to study the relation of surrogate markers to the development of atherosclerosis. These noninvasive imaging modalities include: (1) carotid artery, coronary, and aorta imaging; (2) left ventricular echocardiography imaging; (3) electron-beam computed tomography; (4) magnetic resonance imaging; and (5) ankle-brachial index. Because the incidence of coronary artery disease is a function of the development and progression of atherosclerosis, the use of noninvasive surrogate markers of atherosclerosis can aid in the diagnosis of cardiovascular disease through the identification of subclinical disease. Noninvasive imaging techniques provide an approach for identifying high-risk individuals who may benefit from active intervention to prevent clinical disease. PMID- 11900719 TI - The impact of the National Cholesterol Education Program Adult Treatment Panel III guidelines on drug development. AB - In the newest guidelines of the National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III, more intensive low-density lipoprotein cholesterol-lowering therapy, together with more attention to other lipid and lipoprotein parameters, are recommended for a larger group of dyslipidemic patients than was covered under ATP I and ATP II. A discussion to evaluate how future drug development might be affected by these new guidelines took place at the 14th International Symposium on Drugs Affecting Lipid Metabolism (DALM) conference, held in New York in September 2001. These discussions involved how to develop new lipid-lowering drugs in an era in which so much compelling evidence demonstrates the benefits of statins. Also covered were issues related to the development of drugs with triglyceride indications and whether the proportion of patients achieving NCEP guidelines should be included in the label of lipid lowering drugs. Additional topics discussed included: (1) the possibility of incorporating a non-high-density lipoprotein cholesterol (HDL-C) indication for lipid-lowering drugs, (2) the possibility of obtaining indications for lipid lowering drugs specifically in patients with diabetes, (3) the place of combination lipid-lowering drug therapy in drug development, and (4) whether drugs could be approved to increase levels of HDL-C in patients with isolated low HDL-C. PMID- 11900721 TI - Naming tools and animals: asymmetries observed during direct electrical cortical stimulation. AB - Semantically bounded disorders of verbal processing that result in selective dysnomias for items belonging to specific semantic categories have been well documented in lesion studies. Most commonly dissociations between the categories of living versus non-living things have been reported. Processing of living things such as animals seems to be impaired after bilateral lesions, whereas lesions resulting in impairment of processing of non-living things such as tools seem to be restricted to the left hemisphere. In this study, we tested the naming capabilities of epilepsy patients with subdural electrodes implanted for localization of the epileptogenic zone and preoperative mapping of cognitive functions. Tool and animal items were used, and the results show that during stimulation of the left hemisphere dysnomias for tool items were more pronounced than for animal items. This asymmetry is discussed within a model of more widely bilaterally distributed processing of living category members as compared to more restricted left-sided representation of non-living category members. PMID- 11900722 TI - No deficits at the point of hemispheric indecision. AB - This study attempted to replicate a recent finding by Crow et al. [Neuropsychologia 36 (1998) 1275] showing that about equal skill of right and left hand (i.e. hemispheric indecision) is associated with deficits in cognitive and scholastic achievement. The present study assessed hemispheric indecision by using Annett's [Left, Right, Hand and Brain: The Right Shift Theory, Lawrence Erlbaum, London, 1985] peg moving test and by assessing the consistency of hand preference at school entrance. Non-verbal intelligence, reading and spelling accuracy were assessed about three years later. The sample consisted of 530 boys. Contrary to Crow et al., children with about equal hand skill did not show deficits in non-verbal intelligence, reading and spelling. Also, there were no deficits when inconsistent hand preference was taken as indication of hemispheric indecision. The findings cast doubt on the hemispheric indecision hypothesis and speak specifically against Orton's [Reading, Writing and Speech Problems in Children, Norton, New York, 1937] position, revived by Crow et al., that delayed or absent hemispheric dominance may lead to difficulties with the acquisition of reading and spelling. PMID- 11900720 TI - Managing dyslipidemia in the high-risk patient. AB - Lipid-lowering agents have been shown to reduce morbidity and mortality associated with coronary artery disease (CAD) in all patients. However, these agents are more cost-effective in high-risk patients whose absolute risk of CAD is greater than that of low-risk patients. Furthermore, from preliminary data, it appears that there is greater risk reduction in those subjects achieving lower low-density lipoprotein cholesterol (LDL-C) levels (ie, lower is better). The identification and aggressive treatment of these patients should therefore be a high priority for clinicians. Guidelines from medical organizations, such as the Adult Treatment Panel (ATP) III of the US National Cholesterol Education Program (NCEP), emphasize that patients with CAD, diabetes, or global risk of CAD >20% over 10 years and LDL-C levels >130 mg/dL should receive drug therapy with a goal of reducing LDL-C levels to <100 mg/dL. The recent results of the United Kingdom's Heart Protection Study (HPS) strongly suggest that even those with CAD or who are at high risk and LDL-C levels >100 mg/dL would benefit from drug therapy. Although optimal LDL-C levels have been set at <100 mg/dL for high-risk patients, recent studies show only about 20% of such patients meet these goals. Thus, a large treatment gap remains that needs to be overcome if we are to continue to make significant inroads into preventing further morbidity and mortality in these high-risk subjects. Of therapeutic options available currently and for the near future, statins remain the most effective and well-tolerated form of lipid-lowering therapy. Other therapies include bile acid sequestrants, niacin, and plant stanols. However, none of these is, in general, sufficiently effective as an initial agent to achieve these more aggressive LDL-C goals in the high-risk patient. However, combination therapy with a statin and 1 of these other lipid-lowering agents is useful in patients who are unable to achieve lipid goals on monotherapy. A number of agents for reducing LDL-C levels currently in development may be available in the near future, including 2 new statins: pitavastatin and rosuvastatin. Rosuvastatin, which is in the later stages of the US Food and Drug Administration (FDA) approval process, has been shown to produce significantly greater reductions in LDL-C levels compared with atorvastatin, simvastatin, and pravastatin, and allows more patients to meet lipid goals. Ezetimibe, the first of an entirely new class of LDL-C-lowering agents that inhibit intestinal cholesterol absorption, also appears to offer significant therapeutic value. It is anticipated that these new options will allow clinicians to optimize the management of dyslipidemia in high-risk patients, thereby further reducing the morbidity and mortality of CAD. PMID- 11900723 TI - Manual responses and saccades in chronic and recovered hemispatial neglect: a study using visual search. AB - Hemispatial neglect affects both the ability to respond to targets on the contralesional side of space and to programme saccades to such targets. In the current study, we looked in detail at saccade programming and manual reaction times (RTs) in a range of visual search tasks, in which task difficulty was systematically increased by changing the nature of the distractors. In condition 1, the target was presented with no distractors. In the other conditions, displays contained three distractors that were changed across conditions to manipulate similarity to the target and so task difficulty. We tested two neglect patients, one chronic, one recovered along with two RCVA control patients and 12 age-matched controls. Both neglect patients studied could successfully execute saccades into the neglected field when the target was presented alone. However, a dissociation emerged between the two patients when the target was presented with distractor items. Patient ERs first saccade to target performance in the three search conditions revealed clear effects of distractor type. In contrast for the recovered patient AF, the left/right difference was present for all search displays and appeared to be constant regardless of distractor type. This differential pattern of behaviour may reflect the different underlying neural causes of the neglect in these patients. In the current study, the measurement of saccades allowed the task to be fractionated, and thus, reveal the action of multiple mechanisms controlling saccades in search. PMID- 11900724 TI - Ameliorating neglect with prism adaptation: visuo-manual and visuo-verbal measures. AB - Previous studies have shown that adaptation to rightward displacing prisms improves performance of neglect patients on visuo-manual (VM) tasks such as line cancellation, figure copying, and line bisection [Nature 395 (1998) 166]. The present study further evaluated the effect of prism adaptation (PA) on neglect symptoms by investigating: (a) the range of beneficial effects on common visuo spatial deficits as well as less frequent phenomena like neglect dyslexia; (b) the duration of improvement following a single exposure to the right optical deviation; (c) the extent to which visuo-spatial performance can be comparatively ameliorated in VM tasks and visuo-verbal (VV) tasks (i.e. involving or not the adapted arm, respectively) and (d) the presence and duration of the manual visuo motor bias induced by the prismatic adaptation (i.e. the after-effect). We investigated these issues in a group of neglect patients with right hemispheric damage who were also affected by neglect dyslexia. Following a single, brief prismatic adaptation the results showed that (a) several visuo-spatial abilities, including accuracy in reading single words and non-words, considerably improved, (b) the amelioration was long-lasting, continuing for at least 24h, (c) the presence, amount, and duration of neglect amelioration was not limited to VM tasks, but extended to VV tasks and (d) the presence and duration of the after effect induced by prismatic adaptation remarkably paralleled the presence and duration of the improvement of neglect symptoms. These findings clearly demonstrate that beneficial effects induced by a single PA are very long-lasting and spread over a wide range of visuo-spatial deficits, independent of the type of response required. In addition, our results strongly suggest that the process of adaptation, as revealed by the presence of a visuo-motor after-effect, might be essential for establishing amelioration. In light of its characteristics, the prismatic adaptation technique should be a priority tool for the rehabilitation of the multifaceted hemispatial neglect syndrome. PMID- 11900725 TI - Picture the difference: electrophysiological investigations of picture processing in the two cerebral hemispheres. AB - The nature of semantic memory and the role of the two cerebral hemispheres in meaning processing were examined using event-related brain potentials (ERPs) elicited by pictures in sentences. Participants read sentence pairs ending with the lateralized presentation of three target types: (1) expected pictures, (2) unexpected pictures from the expected semantic category, and (3) unexpected pictures from an unexpected category. ERPs to contextually unexpected pictures were more negative 350-500ms (larger N400s) than those to expected pictures in both visual fields. However, while N400s to the two types of unexpected items did not differ with left visual field presentations, they were smaller to the unexpected items from the expected category with right visual field presentations. This pattern, previously observed to words [Brain Language 62 (1998) 149], suggests general differences in how the two hemispheres use context on-line. Other aspects of the N400 response-and effects on earlier ERP components reveal differences between pictures and words, suggesting that semantic memory access is not modality-independent. The P2 component varied with ending type for right but not left visual field presentations, suggesting that the left hemisphere may use contextual information to prepare for the visual analysis of upcoming stimuli. Furthermore, there was clear evidence for an earlier negativity ("N300"), which varied with ending type but, unlike the N400, was unaffected by visual field of presentation. Overall, the results support our hypothesis that the left hemisphere actively uses top-down information to preactivate perceptual and semantic features of upcoming stimuli, while the right hemisphere adopts a "wait and see" integrative approach. PMID- 11900726 TI - Differential involvement of the hippocampus and temporal lobe cortices in rapid and slow learning of new semantic information. AB - The present study examined the rapid and slow acquisition of new semantic information by two patients with differing brain pathology. A partial double dissociation was found between the patterns of new learning shown by these two patients. Rapid acquisition was impaired in a patient (YR) who had relatively selective hippocampal damage, but it was unimpaired in another patient (JL) who, according to structural MRI, had an intact hippocampus but damage to anterolateral temporal cortex accompanied by epileptic seizures. Slow acquisition was impaired in both patients, but was impaired to a much greater extent in JL. The dissociation suggests that the mechanisms underlying rapid and slow acquisition of new semantic information are at least partially separable. The findings indicate that rapid acquisition of semantic, as well as episodic information, is critically dependent on the hippocampus. However, they suggest that hippocampal processing is less important for the gradual acquisition of semantic information through repeated exposure, although it is probably necessary for normal levels of such learning to be achieved. PMID- 11900728 TI - Electrophysiological estimates of the time course of semantic and phonological encoding during listening and naming. AB - Current psycholinguistic models suggest that we know what we want to say before we decide how we are going to say it: in other words, for speaking, word meaning is activated prior to information about syntax and phonology. Listening likely involves the reverse order of processes: phonological processing before meaning activation. We examined the relative time courses of phonological and semantic processing during language production and comprehension using event-related brain potentials (ERPs). Participants viewed a series of pictures (with the instruction to covertly name the depicted item), or heard a series of words, and made dual choice Go/noGo decisions based on each item's conceptual (whether the item was an animal or an object) and phonological features (whether the item's German name started with a vowel or a consonant). During picture naming, the N200 component (related to response inhibition) indicated that conceptual processing preceded phonological processing by about 170ms. During auditory word processing, on the other hand, the brain activity related to these two aspects of comprehension indicated some temporal overlap with the N200 to phonological processing preceding that to semantic processing by only about 85ms. In sum, the data are compatible with current psycholinguistic models of speech production and comprehension and argue for serial or widely spaced cascaded processing during production but more parallel processing of information during comprehension. PMID- 11900727 TI - A left visual field advantage in perception of gaze direction. AB - Previous work has found a left visual field (LVF) advantage for various judgements on faces, including identity and emotional expression. This has been related to possible right-hemisphere specialisation for face processing, and it has been proposed that this might reflect configural processing. We sought to determine whether a similar LVF advantage may also exist for gaze perception. In two experiments, normal adult subjects made judgements for seen gaze direction (left, right or straight). To assess how visual field may influence perception of gaze direction, eye stimuli were briefly presented unilaterally or bilaterally. In the latter case, the gaze direction of the two seen eyes could be congruent or incongruent (i.e. the two eyes could gaze in the same or different directions). For unilateral displays, performance was more accurate for LVF stimuli than RVF. With bilateral incongruent gaze, the LVF eye influenced judgements more strongly than the RVF eye. No such LVF advantage was found in a control experiment, in which subjects judged pupil size for similar eye stimuli. Taken together, these results reveal a LVF advantage for perception of gaze direction. Since only the eye region was visible, our results cannot be due to a LVF bias in processing the entire face context. Instead they suggest lateralisation specifically in processing the direction of seen gaze. PMID- 11900729 TI - Motor deficits cannot explain impaired cognitive associative learning in cerebellar patients. AB - There is a strong evidence that the cerebellum is involved in associative motor learning. The exact role of the cerebellum in motor learning, and whether it is involved in cognitive learning processes too, are still controversially discussed topics. A common problem of assessing cognitive capabilities of cerebellar patients is the existence of additional motor demands in all cognitive tests. Even if the patients are able to cope well with the motor requirements of the task, their performance could still involve compensating strategies which cost them more attentional resources than the normal controls. To investigate such interaction effects of cognitive and motor demands in cerebellar patients, we conducted a cognitive associative learning paradigm and varied systematically the motor demands and the cognitive requirements of the task. Nine patients with isolated cerebellar disease and nine matched healthy controls had to learn the association between pairs of color squares, presented centrally on a computer monitor together with a left or right answer button. In the simple motor condition, the answer button had to be pressed once and in the difficult condition three times. We measured the decision times and evaluated the correctly named associations after the test was completed. The cerebellar subjects showed a learning deficit, compared to the normal controls. However, this deficit was independent of the motor difficulty of the task. The cerebellum seems to contribute to motor-independent processes, which are generally involved in associative learning. PMID- 11900730 TI - Speechreading circuits in people born deaf. AB - In hearing people, silent speechreading generates bilateral activation in superior temporal regions specialised for the perception of auditory speech [Science 276 (1997) 593; Neuroreport 11 (2000) 1729; Proceedings of the Royal Society London B 268 (2001) 451]. In the present study, FMRI data were collected from deaf and hearing volunteers while they speechread numbers and during a control task in which they counted nonsense mouth movements (gurns). Brain activation for silent speechreading in oral deaf participants was found primarily in posterior cingulate cortex and hippocampal/lingual gyri. In contrast to the pattern observed in the hearing group, deaf participants showed no speechreading specific activation in left lateral temporal regions. These data suggest that acoustic experience shapes the functional circuits for analysing speech. We speculate on the functional role, the posterior cingulate gyrus may play in speechreading by profoundly congenitally deaf people. PMID- 11900731 TI - Functional cerebral asymmetries during the menstrual cycle: a cross-sectional and longitudinal analysis. AB - This study aims at answering two basic questions regarding the mechanisms with which hormones modulate functional cerebral asymmetries. Which steroids or gonadotropins fluctuating during the menstrual cycle affect perceptual asymmetries? Can these effects be demonstrated in a cross-sectional (follicular and midluteal cycle phases analyzed) and a longitudinal design, in which the continuous hormone and asymmetry fluctuations were measured over a time course of 6 weeks? To answer these questions, 12 spontaneously cycling right-handed women participated in an experiment in which their levels of progesterone, estradiol, testosterone, LH, and FSH were assessed every 3 days by blood-sample based radioimmunoassays (RIAs). At the same points in time their asymmetries were analyzed with visual half-field (VHF) techniques using a lexical decision, a figure recognition, and a face discrimination task. Both cross-sectional and longitudinal analyzes showed that an increase of progesterone is related to a reduction in asymmetries in a figure recognition task by increasing the performance of the left-hemisphere which is less specialized for this task. Cross sectionally, estradiol was shown to have significant relationships to the accuracy and the response speed of both hemispheres. However, since these effects were in the same direction, asymmetry was not affected. This was not the case in the longitudinal design, where estradiol affected the asymmetry in the lexical decision and the figural comparison task. Overall, these data show that hormonal fluctuations within the menstrual cycle have important impacts on functional cerebral asymmetries. The effect of progesterone was highly reliable and could be shown in both analysis schemes. By contrast, estradiol mainly, but not exclusively, affected both hemispheres in the same direction. PMID- 11900733 TI - Interhemispheric sensorimotor integration in pointing movements: a study on dyslexic adults. AB - In addition to reading disorders, numerous deficits have been found to be associated with dyslexia, suggesting that various neurological factors might be involved in its etiology. In the present study, we focused on three of the deficits which have been thought to accompany and to a certain extent, to explain dyslexia: an abnormal pattern of hemispheric asymmetry, abnormal hemispheric communication, and abnormal motor control. The aim of the present study was to determine whether adults with reading difficulties perform differently from control subjects in a visuo-manual pointing task, in which the subject was required to point with the right or the left hand to targets appearing to the right or left of a central fixation point. A total of 14 dyslexic adults and 14 control adults participated in this experiment. Motor control was assessed based on the time taken to perform the pointing task, hemispheric asymmetry in terms of the inter-hand differences in the reaction and movement times, and hemispheric communication based on the interhemispheric transfer time under crossed conditions, where the hand and the target were not on the same side. The results showed that neither hemispheric asymmetry nor interhemispheric transfer differed between dyslexic and control adults. However, the dyslexics were significantly slower when performing the pointing task. These results are in agreement with the hypothesis that dyslexia may involve a mild motor deficit. PMID- 11900734 TI - Are central executive functions working in patients with focal frontal lesions? AB - The aim of this study was to examine the hypothesis of a link between frontal cortex and two executive functions in working memory: the capacity to perform a dual task and the ability to inhibit irrelevant information. A dual task designed to assess the capacity to perform storage and processing simultaneously and a directed forgetting task designed to assess the capacity to actively inhibit no longer relevant information were administered to a group of patients with focal frontal lesions and to a group of control participants. The results revealed that despite showing reduced short-term storage, frontal patients performed the dual task and inhibited the no-longer relevant information as well as control participants. These findings suggest that not all-executive processes are exclusively sustained by the frontal cortex [Quart J Exp Psychol 9 (1996) 5; Curr Opin Neurobiol 10 (2000) 195; Neuropsychology (1994) 544; The Cognitive Neuropsychology of Alzheimer-type dementia. Oxford, New York: Oxford University Press, 1996]. PMID- 11900735 TI - Better preservation of memory span relative to supraspan immediate recall in Alzheimer's disease. AB - It is suggested that Alzheimer's disease (AD) patients are able to recall more items on the digit span task than on immediate free recall from a supraspan word list. Two experiments were undertaken to verify this assertion and to understand the basis of the putative span/supraspan discrepancy. The first experiment, involving 35 mildly or moderately demented AD patients, confirmed that digit span significantly exceeded immediate recall from a 10-item supraspan word list. Although digit span also exceeded supraspan recall in 38 elderly non-demented control subjects, the discrepancy was significantly greater within the AD group. In a second experiment, 19 AD cases and 20 controls were assessed with a word span task that used nouns matched by frequency and word length to nouns on the supraspan task. The magnitude of the span/supraspan discrepancy was reduced, indicating that part of the initial discrepancy was due to differences in stimulus items (digits versus common nouns). As before, AD subjects recalled more words on the span task than the supraspan task. However, in striking contrast, NC subjects recalled more words on the supraspan task, further indicating that AD patients are particularly impaired on supraspan recall. Using combined data from 106 subjects in both experiments, digit span performance correlated significantly with supraspan recall for NC but not AD subjects. Moreover, within the AD group the magnitude of the discrepancy was inversely related to a working memory measure derived from the backward digit span. The magnitude of the span/supraspan discrepancy correctly classified 88% of patients with mild dementia and 74% of controls. Results indicate that AD patients are specifically vulnerable to information overload inherent in the supraspan task, a view consistent with the perspective that AD is characterized by prominent disturbances in working memory. PMID- 11900737 TI - Retrieval-induced forgetting in Alzheimer's disease. AB - It is claimed that Alzheimer's disease (AD) patients show reduced inhibitory processing and this has been put forward as a reason why AD patients make intrusion errors at recall. However, the evidence to date has been equivocal, because non-inhibitory mechanisms can account for the pattern of findings. Recently, however, a paradigm has been developed that is claimed to give a purer measure of inhibitory processing in episodic memory, the retrieval-induced forgetting (RIF) paradigm [Inhibitory Processes in Attention, Memory and Language, Academic Press, San Diego, 1994, p. 265; J. Exp. Psychol.: Learning, Memory Cognition 20 (1994) 1063; Psychol. Rev. 102 (1995) 68]. Thus, we were interested whether AD patients would show a deficit in inhibitory processing using this procedure. Participants studied lists of category cue-exemplar pairs (e.g. fruit-orange) then practised retrieval for a subset of items from a subset of categories before taking a final memory test for all studied items. As in previous work, inhibition was measured as the difference between final memory performance for unpractised items from practised categories, and unpractised items from unpractised categories. The results show that AD patients showed normal levels of inhibition with both tests of cued recall and category generation (CG). This suggests that a deficit in inhibitory processes during retrieval is not behind the high levels of intrusion errors made in recall in AD. PMID- 11900732 TI - Modulation of spatial attention by fear-conditioned stimuli: an event-related fMRI study. AB - Stimuli that signal threat can capture subjects' attention, leading to more efficient detection of, and faster responses to, events occurring in that part of the environment. In the present study we explored the behavioural and anatomical correlates of the modulation of spatial attention by emotion using a fear conditioning paradigm, combined with a covert spatial orienting task. Reaction times for the detection of a peripheral target, which was preceded by brief (50ms) presentations of the visual conditioned stimulus (CS+) in either the same or opposite visual field, showed an interaction between stimulus emotionality and attention shifts. We used event-related functional magnetic resonance imaging (fMRI) to characterise the associated neural responses. Consistent with previous studies, conditioning-induced enhanced responses were observed in the amygdala and extrastriate visual cortex. The modulation of spatial attention by a conditioned stimulus was associated with enhanced activity in regions of frontal and parietal cortices previously implicated in spatial attention, as well as in the lateral orbitofrontal cortex (lOFC). PMID- 11900736 TI - Temporal order memory in patients with Korsakoff's syndrome and medial temporal amnesia. AB - Two groups of patients with global amnesia resulting either from Korsakoff's syndrome (KS) or from medial temporal lobe (MTL) damage were compared with groups of matched healthy control subjects on a list discrimination paradigm. Item recognition memory was matched across the amnesic and control groups in order to determine whether KS, but not MTL amnesics are disproportionately impaired on list discrimination as predicted by Parkin's [Functional significance of etiological factors in human amnesia. In: Squire LR, Butters N, editors. Neuropsychology of memory, 2nd ed. New York: The Guilford Press, 1992] hypothesis. However, both patient groups were impaired disproportionately on the temporal order memory task, which is inconsistent with Parkin's hypothesis. It remains possible that the KS patients are more disproportionately impaired than those with MTL damage because both patient groups performed at floor on the list discrimination task. The results are consistent with theories that postulate a critical role for the hippocampus in the kind of associative memory which underlies memory for temporal order, but not in recognition of single items or arbitrary associations between items of similar kinds. PMID- 11900738 TI - Processing pitch and duration in music reading: a RT-ERP study. AB - The originality of this study is to examine the processing of pitch and duration in music reading, using both electrophysiological and behavioral methods. Specifically, it was of interest to determine whether pitch and duration in music reading are processed independently or jointly. A probe, comprising a key and time signature was presented, and participants were required to compute the tonic and/or the best fitting duration. A target note followed the probe and participants made a match/mismatch judgment on the dimension they were required to analyze (i.e. pitch or duration). We hypothesized that, if pitch and duration are processed independently results should show no interference of the irrelevant dimension on the relevant. Results showed that early differences emerged in the ERPs as a function of the task to be performed on the target in block 1. Moreover, RTs were shorter in the pitch than in the duration task and for congruous than incongruous targets. In the ERPs, this congruity effect was reflected by a negative component, to incongruous targets. Most importantly, the congruency of the target note on the irrelevant dimension did not have any effect on the ERPs, suggesting that pitch and duration are processed independently. PMID- 11900739 TI - MRI brain scan analyses and neuropsychological profiles of nine patients with persisting unilateral neglect. AB - Systematic individual neuroanatomical (MRI) and neuropsychological investigations were conducted for nine patients with unilateral neglect persisting at least 3 months after a cerebral vascular accident. The pattern of referrals, together with subsequent investigation, demonstrates that persisting neglect is rare in both right- and left-hemispheric lesioned patients. But while persisting neglect following a left-hemispheric lesion is even rarer than following a right hemispheric lesion, it does occur. The neuroanatomical results indicate that persisting neglect may be associated with a different pattern of damage from acute neglect. In the nine patients investigated, persisting neglect reflected extensive lesions that involved three or more cortical lobes or subcortical regions. The results support previous findings that parietal lesions are common but not essential for persisting neglect. In the seven of nine neglect patients with parietal lesions, the rostral inferior parietal lobe and the parietal frontal junction were involved. Of note was the finding that the brain regions most commonly implicated were the basal ganglia and the superior dorsolateral prefrontal cortex (including the frontal eye field). All of the patients with persisting neglect had a range of neuropsychological deficits, including extinction, personal neglect, and anosognosia for one or more aspects of their neglect. Although it was not possible to demonstrate a double dissociation with this pattern of results, the findings indicate that extinction and anosognosia are dissociable into function-specific forms. Most of the neglect patients also had sustained attention deficits, visual memory problems, and visuospatial constructional difficulties. PMID- 11900740 TI - Odor discrimination in patients with frontal lobe damage and Korsakoff's syndrome. AB - The involvement of the frontal cortex and thalamic nucleus in odor discrimination in humans was assessed. Six patients with frontal lobe brain damage, seven patients with alcoholic Korsakoff's syndrome and 16 healthy comparison subjects completed odor detection and odor discrimination tasks. Multivariate general linear modeling with age as a covariant revealed significantly decreased odor discrimination ability in frontal lobe damaged patients and marginally decreased odor discrimination ability in Korsakoff's syndrome patients as compared to the healthy comparison subjects. No deficits were found in odor detection ability. The findings suggest that in human odor discrimination, there is more involvement of cortico-cortical pathways than of thalamo-cortical pathways. PMID- 11900741 TI - Priming summation in the cerebral hemispheres: evidence from semantically convergent and semantically divergent primes. AB - The ability to activate and to maintain a large and relatively undifferentiated semantic field has been thought to be an important component of lexical semantic processing by the right hemisphere (RH). An implication of this unique propensity of the RH was examined in the present study that included two divided visual field priming experiments with SOAs of 800 and 2500 ms. The experiments investigated the ability of the RH and the left hemisphere (LH) to summate activation from multiple primes followed by a laterally presented ambiguous target word. The priming words either converged onto the same semantic representation (i.e. all three words related to either the dominant or to the subordinate meaning of the target) or diverged onto distinct semantic representations (i.e. two words related to the dominant and one to the subordinate meaning of the target, or vice versa). Results indicated that for either an 800 or 2500 ms stimulus onset asynchrony (SOA) the LH benefited most from three semantically convergent primes that converged onto the dominant meaning of the ambiguous target word. There was no facilitation when three subordinate primes preceded the target. When the primes diverged onto different meanings, there was significant facilitation for the 800 ms SOA only. In contrast, with an 800 ms SOA, the RH benefited only from semantically divergent primes, that diverged onto alternate meanings of the ambiguous target word. With a 2500 ms SOA, the RH benefited from all combinations of primes. The discussion focuses on the implications for language processing of the differences between the two hemispheres in the scope and temporal pattern of the multiple prime effect. PMID- 11900742 TI - Disturbed visual processing contributes to impaired reading in Alzheimer's disease. AB - The relationship between visual processing dysfunction and oral reading impairment was investigated in 17 patients with probable or possible Alzheimer's disease (AD). When dementia severity was controlled, a significant relationship was found between single word oral reading impairments and difficulties discriminating words written in different fonts and photographs of objects in different orientations, which are all functions believed to be dependent on the integrity of left ventral temporal-occipital visual association regions. By contrast, there was no significant relationship between reading performance and the score on a test of spatial localization, believed to be more dependent on parietal lobe function. There was also no relationship between reading ability and discrimination of unfamiliar faces, a function thought to engage right inferotemporal lobe structures. In contrast to the significant association between impaired reading and certain visual processes, when dementia severity was controlled, there was no relationship between reading and lexical semantic impairment. These results highlight the contribution of visual processing deficits to impaired oral reading in AD and further suggest that this association may derive from neuropathological changes in areas of the left temporal occipital lobes specialized for high-level visual processing. PMID- 11900743 TI - A sex difference in reliance on vision during manual sequencing tasks. AB - The manual praxis system is a left-hemisphere based motor programming system that is involved in the selection of hand and arm movements and is particularly important for the control of movements made with minimal external sensory guidance. Few studies have explored the parameters surrounding normal praxic function, but preliminary data suggest that men may rely less heavily on praxic control than women. To further investigate this possibility, we tested healthy individuals on two tasks designed to recruit the praxis system and on a visually guided control measure. Participants performed each task with vision and without. We hypothesized that, if men are less fully reliant on the praxis system than women, then their performance should be more adversely affected by the removal of vision on tasks that are ostensibly praxic. Consistent with this prediction, we found a significantly larger drop in men's scores than in women's when vision was occluded. Importantly, this pattern was selective to the praxic measures. In contrast, men were no more impaired by the loss of vision than were women on an externally-guided pegboard task. These findings may reflect an anatomical sex difference in the organization of the praxis system within the left cerebral hemisphere. PMID- 11900744 TI - Mental rotation versus invariant features in object perception from different viewpoints: an fMRI study. AB - It has been proposed that object perception can proceed through different routes, which can be situated on a continuum ranging from complete viewpoint-dependency to complete viewpoint-independency, depending on the objects and the task at hand. Although these different routes have been extensively demonstrated on the behavioral level, the corresponding distinction in the underlying neural substrate has not received the same attention. Our goal was to disentangle, on the behavioral and the neurofunctional level, a process associated with extreme viewpoint-dependency, i.e. mental rotation, and a process associated with extreme viewpoint-independency, i.e. the use of viewpoint-invariant, diagnostic features. Two sets of 3-D block figures were created that either differed in handedness (original versus mirrored) or in the angles joining the block components (orthogonal versus skewed). Behavioral measures on a same-different judgment task were predicted to be dependent on viewpoint in the rotation condition (same versus mirrored), but not in the invariance condition (same angles versus different angles). Six subjects participated in an fMRI experiment while presented with both conditions in alternating blocks. Both reaction times and accuracy confirmed the predicted dissociation between the two conditions. Neurofunctional results indicate that all cortical areas activated in the invariance condition were also activated in the rotation condition. Parietal areas were more activated than occipito-temporal areas in the rotation condition, while this pattern was reversed in the invariance condition. Furthermore, some areas were activated uniquely by the rotation condition, probably reflecting the additional processes apparent in the behavioral response patterns. PMID- 11900745 TI - Neurophysiological evidence for two processing times for visual object identification. AB - Event-related brain potentials (ERPs) were recorded to fragmented pictures of objects that were named correctly or were not to investigate the time course of visual object identification. The first ERP difference distinguishing identified from unidentified pictures estimates the upper limit of the time by which human brain regions have begun to activate long-term memory (LTM) representations specifying the identity of a visual object. Data from 15 young adults indicate that this time varies with the extent to which object parts are recoverable from the visual input, being approximately 200 ms earlier with recoverable than unrecoverable parts. Successful identification is evident by approximately 300 ms when object parts and overall structural configuration are readily recoverable but not until approximately 550 ms when object parts are difficult or impossible to recover (i.e. too poorly specified by the available contours to be recovered). In both cases, successful identification is associated with greater relative positivity. However, unidentified recoverable pictures are associated with an enhanced frontal negativity (N350), linked to object matching operations, not seen for non-recoverable pictures. Taken together, these results implicate two distinct processing sequences in the successful identification of visual objects. PMID- 11900746 TI - Relationship between attentional performance and corpus callosum morphometry in patients with Alzheimer's disease. AB - There has been considerable interest in cognitive deficits associated with Alzheimer's disease (AD) and relationships between these impairments and specific cortical atrophies. Two previous studies [Neuropsychologia 28 (1990) 1197; Dementia 3 (1992) 350] have found that AD patients exhibit significant impairments in the attentional ID/ED set-shifting tasks of the CANTAB battery which involved attentional shifting abilities. But, at present, no study has examined the neural bases of these abilities in AD patients. In the present study, the relationship between performances on this attentional test and morphometry of the anterior and posterior corpus callosum is examined in AD patients in the mild to moderate stages of the disease (n=30, mean age=74.1+/-4.9 years, mean MMSE score=23.9+/-2.6). A control group is constituted (n=20, mean age=73.15+/-5.5 years) for comparison of cerebral measurements. The stepwise multiple regression analysis indicates that the relative contribution for the total callosal and the anterior CC areas of the simple discrimination subtest is significantly positive whereas for the posterior callosal areas the relative contribution of the more complex subtest is significantly positive. AD patients from the subgroup "low", who failed to do the nine subtests of the attentional set-shifting tasks, exhibit smaller callosal areas than control subjects. There is no significant difference for all callosal measurements between AD patients from the subgroup "high", who completely succeeded the test, and control subjects. Our findings suggest that the anterior corpus callosum would be related to attentional shifting abilities in AD patients. Moreover, these patients with probable AD seem heterogeneous for performances in the attentional test of the CANTAB and for callosal atrophies. PMID- 11900747 TI - Hemispheric differences in global and local processing with orientation classification tasks. AB - Hemispheric differences in global and local processing were examined in one experiment with hierarchical stimuli. The figures consisted of large squares with the right or left side missing made up of small squares with the right or left side missing. The subjects were asked to decide the opening (left/right) of the square either at the global level or at the local level. The findings showed that with the task and stimuli used here global judgements were as fast and accurate as local judgements, the interference was bidirectional and symmetrical and, finally, that the right hemisphere and the left hemisphere had the same ability to manage with global and local information. So, the experiment does not provide evidence for hemispheric specialisation in global and local processing. PMID- 11900748 TI - Long-term memory and levels-of-processing in autism. AB - Prior studies reported that long-term memory (LTM) was basically unimpaired in individuals with autism. However, people with autism have been found to perform worse than ability-matched controls when verbal materials to be remembered are semantically related. In normal subjects, semantic processing of verbal materials facilitates LTM better than 'shallow' (phonological or perceptual) processing, which is known as the levels-of-processing effect. In this study, the relationship between LTM and semantic processing was investigated using a levels of-processing task. In Study 1, a levels-of-processing task was conducted on healthy volunteers, which confirmed the levels-of-processing effect with our task. In study 2, the same task was conducted on autistic subjects with mild or no mental retardation and ability-matched controls. The levels-of-processing effect was confirmed in the control group. Although overall performance in the two groups was comparable, the levels-of-processing effect was not found in the autistic group. LTM resulting from perceptual processing was better in the autistic group than in the control group, indicating superior "rote memory" in individuals with autism. Furthermore, the pattern of correlations between LTM performance and cognitive measures differed greatly between the two groups. The lack of the levels-of-processing effect, which has not been reported in other psychiatric or neuropathological conditions, suggests an abnormal relationship between semantic memory and episodic memory in individuals with autism. PMID- 11900749 TI - Performance on tests of frontal lobe function reflect general intellectual ability. AB - Recent studies have indicated that performance on tests of frontal lobe function are highly associated with general intellectual ability (g). Some authors have even claimed that the available evidence does not support a more specific account of frontal lobe function than to provide a general intellectual function for the performance of goal directed tasks. We examined the relationship between performance on the WAIS-R (as a measure of g) and performance on standard tests of frontal lobe function in 123 healthy individuals. Our results demonstrate that in healthy individuals (i) performance on the most popular tests of frontal lobe function shares significant variance, and (ii) a large proportion of that shared variance is highly associated with performance on the Wechsler Adult Intelligence Scales-Revised (WAIS-R), so that the tests are similar to the extent that they measure g. Performance on the Modified Card Sorting Test (MCST), however, is not related to g. The results support the claim that many tests of frontal lobe function measure primarily a non-specific intellectual function but also indicate that some tests, like the MCST, may be assessing more specific cognitive operations. PMID- 11900750 TI - Hemispheric semantic priming in the single word presentation task. AB - Functional brain asymmetries in semantic activation were studied by presenting categorically related (e.g. TABLE-BED) or unrelated primes and targets to the left visual field (LVF)/right hemisphere or to the right visual field (RVF)/left hemisphere in the single word presentation lexical decision task. The results showed that the primes in the RVF/left hemisphere primed lexical decisions to the targets both in the RVF and in the LVF. However, the primes in the LVF/right hemisphere did not induce any priming in the LVF or RVF. These results suggest that the left hemisphere automatically activates categorically related meanings in both hemispheres. The role of the right hemisphere in automatic semantic processing may be very limited. PMID- 11900751 TI - Memory for perceived and imagined pictures--an event-related potential study. AB - Event-related potentials (ERPs) and behavioural measures were used to investigate recognition memory and source-monitoring judgements about previously perceived and imagined pictures. At study, word labels of common objects were presented. Half of these were followed by a corresponding picture and the other half by an empty frame, signalling to the participants to mentally visualise an image. At test, participants in a source-monitoring task made a three-way discrimination between new words and words corresponding to previously perceived and imagined pictures. Participants in an old/new-recognition task indicated whether test words were previously presented or not. In both tasks, correctly identified old items elicited more positive-going ERPs than correctly judged new items. This widely distributed old/new effect was found to have an earlier onset and to be of a greater magnitude for imagined than for perceived items. Task (source versus item-memory) affected the old/new effects over prefrontal areas and the reaction times to remembered old items. The present findings are consistent with the view that a greater amount, or a different type, of information is necessary for accurate source-memory judgements than for correct recognition, and moreover, that different types of source-specifying information revive at different rates. In addition, the results add weight to the view that the late widespread ERP old/new effect is sensitive to the quality or the amount of information retrieved from memory. PMID- 11900752 TI - Spatial bias: effects of early reading direction on Korean subjects. AB - Spatial bias may occur in subjects performing a number of cognitive and visual motor tasks. These include coordinate visuospatial computations (e.g. bisecting a line) and categorical representations of syntactic information (e.g. drawing a picture depicting the action of a sentence). Readers of European languages scan from left-to-right and this learned visual scanning may contribute to leftward spatial bias. In 30 subjects who first learned to read in a top-to-bottom, right to-left direction (right-left vertical readers, RL), we tested spatial-syntactic bias by reading sentences to subjects, who drew pictures depicting the actions. We noted whether the subject of the sentence was located leftward or rightward of the object. We assessed visual-spatial bias by measuring subjects' accuracy at bisecting lines, and by measuring how closely their drawings on the house-tree person test were centered on the page. On the spatial-syntactic task, the RL were not right- or left-biased (P=0.581). Korean controls (left-right horizontal readers, LR) also showed no significant spatial-syntactic bias. RL only tended to bisect lines leftward, but displaced house-tree-person drawings left of page center (P<0.001). LR erred leftward on line bisection, and had a smaller magnitude leftward bias on drawing tasks. We conclude that a leftward spatial syntactic bias may not be innate and does not appear to be influenced by learned reading direction. In contrast, the leftward visual-spatial bias may occur in subjects whose cultural and reading background is neither western nor left-to right. PMID- 11900753 TI - Perceived gaze direction in faces and spatial attention: a study in patients with parietal damage and unilateral neglect. AB - Perceived gaze in faces is an important social signal that may influence orienting of attention in normal observers. Would such effects of gaze still occur in patients with right parietal damage and left neglect who usually fail to attend to contralesional space? Two experiments tested for effects of perceived gaze on visual extinction. Face or shape stimuli were presented in the right, left, or both hemifields, with faces looking either straight ahead or toward the opposite field. On bilateral trials, patients extinguished a left shape much less often when a concurrent right face looked leftward rather than straight ahead. This occurred, even though gaze was not relevant to the task and processing of facial signals implied attention to a competing ipsilesional stimulus. By contrast, rightward gaze in faces presented on the left side had no effect on extinction, suggesting that gaze cues are not extracted without attention. Two other experiments examined effects of perceived gaze on the detection of peripheral targets. Targets appeared at one of four possible locations to the right or left of a central face looking either toward the target location, another location on the same side, the opposite side, or straight ahead. Face and gaze were not relevant to the task and not predictive of target location. Patients responded faster when the face looked toward the target on both the contralesional and ipsilesional sides. In contralesional space, gaze allowed shifting of attention in a specific quadrant direction, but only to the first target along the scan path when there were different possible locations on the same side. By contrast, in intact ipsilesional space, attention was selectively directed to one among different eccentric locations. Control experiments showed that symbolic arrow cues did not produce similar effects. These results indicate that even though parietal damage causes spatial neglect and impairs the representation of location on the contralesional side, perceived gaze in faces can still trigger automatic shifts of attention in the contralesional direction, suggesting the existence of specific and anatomically distinct attentional mechanisms. PMID- 11900754 TI - Visual extinction with double simultaneous stimulation: what is simultaneous? AB - Patients with unilateral lesions to the parietal lobe leading to extinction were tested with stimuli presented in both hemifields. In addition to simultaneous presentation of two stimuli, trials were given with the contralesional or ipsilesional side having an onset slightly before the other. In the first experiment, subjects were asked to identify the stimuli. In agreement with di Pellegrino et al. [Neuropsychologia 35(1997)1215], all patients showed maximal extinction when the stimuli were physically simultaneous. In the second experiment, the same patients were requested to report whether the left or right item had been presented first. In accordance with Rorden et al. [Neuropsychologia 35(1997)421], all three patients required the contralesional item to have a significant lead in order to in order to be judged as occurring first. Taken together, these findings appear to present a paradoxical picture of extinction: the first task implies that items compete maximally when they are physically simultaneous, while the second study suggests that the physical simultaneity is not phenomenally special to the patients. Attempts to reconcile these findings with popular models of extinction are discussed. PMID- 11900756 TI - The effects of stimulus symmetry on landmark judgments in left and right visual fields. AB - Line bisection impairment is greater following right compared to left hemisphere damage, suggesting that some of the underlying visuo-spatial mechanisms may be lateralised. One important perceptual component of line bisection is estimating stimulus midpoint. Here, we used a modified version of the perceptual landmark task to examine, in healthy individuals, how the midpoint of a stimulus is apprehended, and if the cerebral hemispheres are equally as efficient in performing such a task. We show that the right, relative to the left, hemisphere is both faster and more accurate at apprehending prebisected lines, but no better at apprehending misbisected lines. We then demonstrate that the right hemisphere advantage stems from a specialisation in detecting stimulus symmetry; by associating prebisected lines with the presence of display symmetry, transect location can be inferred without having to explicitly calculate the midpoint of lines. The findings provide a further reason why right hemisphere damage is so deterimental to perceptual line bisection. In addition, the data indicate that the detection of visual symmetry is preferentially lateralised to the right hemisphere. PMID- 11900755 TI - Less developed corpus callosum in dyslexic subjects--a structural MRI study. AB - BACKGROUND: Based on previous studies and due to the characteristics of dyslexia as an auditory phonological decoding disorder, we predicted that the shape of the posterior corpus callosum (CC) would differ between dyslexic and control subjects. METHOD: Twenty right-handed boys with developmental dyslexia were selected from a carefully screened general population sample (mean age 11 years) and compared to a matched control group. The CC contour was manually traced on the aligned midsagittal MR slice and total callosal area and its subregions were compared between the groups. A statistical shape analysis and subsequent CC classification was performed using a recently developed shape model method. RESULTS: The shape analysis revealed shorter CC shape in the dyslexic group, localised in the posterior midbody/isthmus region. This region contains interhemispheric fibers from primary and secondary auditory cortices. A shape length difference larger than a fixed threshold in the posterior midbody region could correctly discriminate between control and dyslexic subject in 78% of the cases, where a dyslexic CC was shorter in this region than a control CC. However, there were no significant group differences with respect to overall CC area or subregions. CONCLUSION: A clear shape difference in the posterior midbody of the CC was found between dyslexic and control subjects. This fits with recent other studies that have reported a strong growth factor in this CC region during the late childhood years, coinciding with literacy acquisition. Our results show that the dyslexic group has not undergone the same growth pattern as the normal reading group. PMID- 11900757 TI - Distractibility after unilateral resections from the frontal and anterior cingulate cortex in humans. AB - The prefrontal and anterior cingulate cortex figure prominently in current models of directed attention. The efficacy of selective attention was studied in a distractibility study carried out with patients who had unilateral surgical excisions from the frontal cortex with or without involvement of the cingulate gyrus, patients with unilateral anterior temporal resections, and normal control subjects. In Task A, participants responded to target stimuli presented at the foveal location in the presence or absence of distracting peripheral stimuli that appeared 200 ms before the target's onset. The presence of a distractor caused a comparable increase in reaction times to targets across all groups. In Task B, a distractor appeared during every trial but it appeared at a new location on 12% of the trials. A comparable transient increase in reaction times was observed in patients with frontal cortical resections that spared the anterior cingulate gyrus, patients with anterior temporal lobe resections, and the normal control group. Thus, lesions restricted to the prefrontal cortex need not impair the ability to attend and respond to a target event in the presence of distraction. Interestingly, no distracting effect was observed during Task B in patients with lesions that invaded the anterior cingulate gyrus, suggesting that this cortical region is involved in responding to or habituating to environmental stimuli. PMID- 11900758 TI - Kinematic optimization of spatiotemporal patterns in paretic arm training with stroke patients. AB - The effect of rhythmic cueing on spatiotemporal control of sequential reaching movements of the paretic arm was studied in 21 hemispheric stroke patients. Reaching movements were studied with and without rhythmic metronome cuing in a counterbalanced design. Metronome frequencies were entrained to the naturally selected frequency of the patient. Results indicate statistically significant (P<0.05) improvements of spatiotemporal arm control during rhythmic entrainment. Variability of timing and reaching trajectories were reduced significantly. Time series analysis of sequential movement repetitions showed an immediate reduction in variability of arm kinematics during rhythmic entrainment within the first two to three repetitions of each trial. Rhythm also produced significant increases in angle ranges of elbow motion (P<0.05). Analysis of acceleration and velocity profiles of the wrist joint showed significant kinematic smoothing during rhythmic cuing. The link between rhythmic sensory timing and spatiotemporal motor control was investigated using a mathematical optimization model with minimization of peak acceleration as criterion. Rhythmically cued acceleration profiles fit the predicted model data significantly closer (P<0.01) than the self paced profiles. Since velocity and acceleration are mathematical derivatives of position-time trajectories, the model data suggest that enhanced timing precision via temporal phase and period coupling of the motor pattern to the rhythmic time timekeeper enhances the brain's computational ability to optimally scale movement parameters across time. PMID- 11900759 TI - A generalized role of interhemispheric interaction under attentionally demanding conditions: evidence from the auditory and tactile modality. AB - The present study investigated whether dividing critical information across the hemispheres in the auditory and tactile modalities aids performance more for computationally complex rather than computationally simpler task-a pattern previously observed in the visual modality [Cortex 26 (1990) 77; Neuropsychology 12 (1998) 380; Neuropsychologia 30 (1992) 923]. We conducted two experiments, one in the auditory and one in the tactile modality, that were analogous to those previously performed in the visual modality. In agreement with previous findings, for both modalities we observed that the performance advantage exhibited for within-hemisphere processing in the computationally simpler condition (that required fewer steps to reach a decision) was diminished in the computationally more complex condition. In the auditory experiment we also manipulated computational complexity by varying the amount of time available for processing information. The within-hemisphere advantage in performance was also significantly reduced when complexity was increased through temporal manipulations. These findings suggest that the brain may use interhemispheric interaction as a general strategy to increase computational resources, independent of sensory modality and the manner in which computational demands are increased. PMID- 11900760 TI - Limitations of attentional orienting. Effects of abrupt visual onsets and offsets on naming two objects in a patient with simultanagnosia. AB - It has been proposed that the underlying deficit for some simultanagnosics is the inability to bilaterally orient attention in space due to parietal damage. In five experiments, we examine the performance of a patient with simultanagnosia secondary to bilateral occipito-parietal lesions, IC, in naming pairs of line drawings. With simultaneous presentation and disappearance of objects (Experiment 1), IC typically named a single object. IC's performance dramatically improved when the two drawings alternated every 500 ms (Experiment 2). This improvement was not due to the abrupt onset of the second drawing "capturing attention", as indicated by the results of Experiment 3. Experiments 4 and 5 demonstrated that the crucial factor in improving IC's performance with simultaneous presentation of visual objects was the offset of one of the two stimuli. We propose that IC's impairment in naming two objects is attributable to the inability to "unlock" attention from the first object detected to other objects in the array. Visual offset of the first object disengages attention from the first object, allowing it to be allocated to the second object. PMID- 11900761 TI - Bilateral vestibular stimulation does not improve visual hemineglect. AB - This work compared the effect of unilateral (right and left) and bilateral vestibular stimulation in a right-brain-damaged patient with neglect. Neglect was improved following left caloric vestibular stimulation, and worsened following right vestibular stimulation. On the other hand, no modification of neglect was observed after bilateral vestibular stimulation. These results support the idea that caloric vestibular stimulation may improve neglect through a specific effect; bilateral stimulation making the putative activation bilateral and symmetrical does not affect the lateral bias of neglect. PMID- 11900762 TI - Ginseng total saponin potentiates acute U-50,488H-induced analgesia and inhibits tolerance to U-50,488H-induced analgesia in mice. AB - In the present study, an attempt has been made to investigate whether the potentiating effect of U-50,488H (U50)-induced analgesia by ginseng total saponin (GTS) is playing a role in inhibiting the tolerance to U50-induced analgesia as measured using the tail-flick test in mice. GTS (100 and 200 mg/kg i.p.), on acute administration, potentiated the U50 (40 mg/kg i.p.)-induced analgesia in U50-naive mice. Twice daily administration of U50 (40 mg/kg i.p.) for 6 days resulted in tolerance to U50-induced analgesia in mice. Chronic administration (Days 4-6) of GTS (50, 100, and 200 mg/kg i.p.) to U50-tolerant mice dose dependently inhibited the tolerance to U50-induced analgesia. On the other hand, chronic administration of GTS (50, 100, and 200 mg/kg i.p.) dose-dependently potentiated the U50-induced analgesia in U50-naive mice. The dose-response curve to U50-induced analgesia in U50-tolerant mice was shifted rightward (2.6-fold) as compared to U50-naive mice, indicating the development of tolerance to U50 induced analgesia. GTS (100 mg/kg i.p. o.d.), on chronic administration, prevented the rightward shift of dose-response curve to U50-induced analgesia in U50-tolerant mice, whereas in U50-naive mice it resulted in leftward shift (0.6 fold). It can be concluded that acute and chronic administration of GTS potentiates the U50-induced analgesia in U50-naive mice. The potentiating effect of GTS on U50-induced analgesia may be partially responsible in the inhibition of tolerance to U50-induced analgesia in mice. PMID- 11900763 TI - Possible anti-Parkinson properties of N-(alpha-linolenoyl) tyrosine: a new molecule. AB - Tyrosine is unable to cross the blood-brain barrier and is therefore unable to improve the status of brain dopamine (DA) and to provide relief for patients with Parkinson's disease (PD) or other DA-insufficient disorders. We report the creation of an amide bond molecule [N-(alpha-linolenoyl)tyrosine (NLT)] that combines tyrosine with a fatty acid mixture. NLT significantly improves the rotational behavior of rats [following unilateral striatal lesions (as a model for Parkinson's)] and overcomes the exaggerated eye-blinking induced by a potent DA-depleting agent (as a model for essential blepharospasm). These results are supported by the finding that NLT's mode of action, in striatum, is the same as the mode of action of D-amphetamine. They both induce an increase in the DA level, DA turnover and release. PMID- 11900764 TI - Male and female C57BL/6 mice respond differently to diazepam challenge in avoidance learning tasks. AB - Benzodiazepines (BZ) impair learning and memory performance of animals. The goal of this study was to examine sex differences in the effects of diazepam on learning and memory of C57BL/6 mice in avoidance paradigms. Male and female C57BL/6 mice were tested in the one-way active avoidance, step-down passive avoidance, and foot-shock pain threshold tasks, following administration of vehicle or diazepam (1 mg/kg). No substantial sex or drug effects on the threshold of the pain response to shock were found. There were no significant differences in avoidance performance between vehicle-treated male and female mice while 1 mg/kg of diazepam produced opposite effects on performance of males and females in both tasks. Diazepam-treated females learned faster in the active avoidance task and showed stronger retention in the passive avoidance task. In contrast, diazepam impaired learning of males in the active avoidance task and had no effect on their performance in the passive avoidance task. Diazepam induced impairment in males was not due to higher sensitivity to the sedative effect of diazepam as females were more sedated than males on the first trial of the passive avoidance task. Our data showed that sedative and amnesic effects of BZs are not tightly linked. This study also suggests that cognitive effects of BZs in rodents could be sex dependent and highlight the importance of using both sexes in studies on behavioral effects of psychoactive drugs. PMID- 11900765 TI - Interaction of angiotensin II and adenosine A1 and A2A receptor ligands on the writhing test in mice. AB - The effects of adenosine A1 and A2A receptor agonists and antagonists administered intraperitoneally (i.p.) and their interaction with angiotensin II (Ang II) administered intracerebroventricularly (i.c.v.) were studied in mice using the acetic acid-induced abdominal constriction test. Ang II (0.1 microg/mouse) induced antinociception in this model. The adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA; 0.05, 0.25 and 0.5 mg/kg) also showed a well-developed antinociceptive effect. Ang II (0.1 microg/mouse) administered 5 min before CPA (0.25 mg/kg) decreased the number of writhes, i.e., it enhanced the antinociceptive effect of CPA. Losartan, an AT1 receptor antagonist (25 microg/mouse i.c.v.), enhanced the antinociceptive effect of CPA, while the AT2 receptor antagonist 1-[-4-(dimethylamino)-3-methylphenylmethyl]-5-diphenylacetyl) 4,5,6,7-tetrahydro 1H-4-imidazol [4,5c]pyridine-6 carboxylic acid, ditrifluoroacetate, dihydrate (PD 123319; 10 microg/mouse) had less effect. 8 Cyclopentyl-1,3-dipropylxanthine (DPCPX; 0.1 mg/kg), an adenosine A1 receptor antagonist, exhibited a pronociceptive effect and did not change the antinociceptive effect of Ang II. The adenosine A2A receptor agonist PD-125944 (DPMA; 0.1, 0.5 and 1 mg/kg) showed pronounced antinociceptive effect. Ang II (0.1 microg/mouse) did not significantly influence the antinociceptive effect of DPMA (0.1 mg/kg). The A2A receptor antagonist 3,7-dimethyl-1-propargilxanthine (DMPX; 0.1 mg/kg) had no effect on the number of writhes and did not influence the effect of Ang II. These data indicate that the antinociceptive effect of Ang II interacts with that produced by adenosine A1 receptor agonist. PMID- 11900766 TI - 5-HT2A receptor-stimulated phosphoinositide hydrolysis in the stimulus effects of hallucinogens. AB - The role of 5-HT2A-mediated stimulation of phosphoinositide hydrolysis in the discriminative effects of hallucinogens was investigated in PC12 cells stably expressing the rat 5-HT2A receptor (PC12-5-HT2A cells). The hallucinogenic compounds, D-lysergic acid diethylamide (LSD), (-)2,5-dimethoxy-4 methylamphetamine (DOM), psilocybin, N,N-dimethyltryptamine (DMT), 5-methoxy-N,N dimethyltryptamine (MDMT) and N,N-diethyltryptamine (DET), all caused a concentration-dependent increase in the generation of [3H]inositol phosphates. The nonhallucinogenic compounds, 6-fluoro-N,N-diethyltryptamine (6-F-DET), lisuride and quipazine, also displayed significant efficacy in stimulating phosphoinositide hydrolysis, while 2-bromo-lysergic acid diethylamide (BOL), which is not a hallucinogen, did not alter inositol phosphate generation. The beta-carbolines, harmaline and harmane, also did not alter phosphoinositide hydrolysis. Comparison of these results with previous drug discrimination studies indicated the apparent lack of correlation between the degree of substitution in LSD- and DOM-trained animals and efficacy in stimulating phosphoinositide hydrolysis. The present study indicates that 5-HT2A-mediated stimulation of phosphoinositide hydrolysis does not appear to be the sole critical signaling mechanism involved in the discriminative effects of hallucinogens. PMID- 11900767 TI - The effect of Morinda officinalis How, a Chinese traditional medicinal plant, on the DRL 72-s schedule in rats and the forced swimming test in mice. AB - The present study observed the antidepressant-like action of the medicinal plant Morinda officinalis in the differential reinforcement of low rate 72-s (DRL 72-s) schedule, a behavioral screen selective and sensitive to antidepressant drugs, and the forced swimming test, a well-known animal model of depression. In the DRL 72-s schedule in rats, the plant extract (25-50 mg/kg), similar to clinically effective antidepressant drug desipramine (5-10 mg/kg), significantly reduced response rate and efficiency ratio while at the same time increasing reinforcement rate. In the forced swimming test in mice, the plant extract (50 mg/kg), like the effect of desipramine (20 mg/kg), also elicited a significant reduction in the duration of immobility. A tendency to this phenomenon could be seen at the dose of 100 mg/kg. Meanwhile, the plant extract, in the effective doses for the forced swimming test, had no effects on spontaneous motor activity in mice. These findings provide further support for the conclusion that M. officinalis extract possesses the antidepressant effect. PMID- 11900768 TI - Cholinergic and dopaminergic mechanisms involved in the recovery of circadian anticipation by aniracetam in aged rats. AB - We have reported that repeated administration of aniracetam (100 mg/kg p.o.) for 7 consecutive days recovers mealtime-associated circadian anticipatory behavior diminished in aged rats. The present study examines the mode of action underlying the restoration by aniracetam with various types of receptor antagonists. Coadministration of scopolamine (0.1 mg/kg i.p.) or haloperidol (0.1 mg/kg i.p.) for the last 3 days significantly reduced the restorative effects of aniracetam without affecting the timed feeding-induced anticipatory behavior by each receptor antagonist itself. The other receptor antagonists, mecamylamine (3 mg/kg i.p.), 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX, 1 microg/rat i.c.v.) had no effect on either the basal or aniracetam-elicited circadian anticipation. In contrast, ketanserin (1 mg/kg i.p.) itself recovered the diminished anticipatory behavior as aniracetam did, but it did not alter the restorative effects of aniracetam. Among the receptor antagonists tested, NBQX reduced appetite and haloperidol induced circadian hypoactivity. These results suggest that the food-entrainable circadian oscillations or the temporal regulatory system of behavior is modulated by cholinergic, dopaminergic and serotonergic systems. Furthermore, aniracetam may restore the aging-diminished behavioral anticipation by activating muscarinic acetylcholine (ACh) and/or dopamine (DA) D2 receptors through the enhanced release of ACh and/or DA in the brain. PMID- 11900769 TI - The reinforcing effects of acetaldehyde in the posterior ventral tegmental area of alcohol-preferring rats. AB - Acetaldehyde (ACD), the first metabolite of ethanol, is a biologically active compound, which may mediate some of the reinforcing, behavioral and neurotoxic effects of ethanol. The objective of this study was to test the hypothesis that ACD is reinforcing within the mesolimbic system. The intracranial self administration (ICSA) technique was employed to determine whether ACD was reinforcing in the posterior ventral tegmental area (VTA), a site that supports the reinforcing actions of ethanol. Adult female alcohol-preferring (P) rats were implanted with guide cannulae aimed at the posterior VTA. Subjects were placed in two-lever operant chambers 7-10 days after surgery. Responding on the "active lever" on a fixed ratio 1 (FR1) schedule of reinforcement caused the delivery of 100 nl of infusate, whereas responses on the "inactive lever" were without consequences. Rats were assigned to one of five groups that self-administered either artificial cerebrospinal fluid (aCSF) throughout all eight sessions (4 h in duration) or 3- and 6-, 11- and 23-, 45- and 90- or 180- and 360-microM ACD for the eight sessions, with the lower concentration of ACD given for the initial four sessions and the higher concentration of ACD given for the last four sessions. A second experiment examined the acquisition (first four sessions), extinction (aCSF in sessions 5 and 6) and reinstatement using 90-microM ACD. A third experiment examined the effects of extending the time-out period (from 5 to 55 s) on the number and pattern of infusions of 23-microM ACD. Adult P rats readily self-administered 6-90-microM ACD and discriminated between the active and inactive levers. Furthermore, rats self-administering 90-microM ACD also demonstrated extinction behavior when aCSF was substituted for ACD and gradually reinstated active lever responding when ACD was reintroduced. P rats maintained similar numbers of infusions and infusion patterns under both time-out schedules. Overall, the data indicate that ACD is a potent reinforcer within the posterior VTA of the P rat. PMID- 11900771 TI - A simple integrated circuit device for measuring distances traveled and determining speed in open-field environments. AB - An easily constructed and inexpensive device for measuring distance and determining mean speed, which utilizes an integrated circuit (IC) photosensor system, is described. It was designed for evaluating activity of small animals in open-field environments, being used in conjunction with a video-recording system. The horizontal locomotor path of the animal is manually tracked directly from the video monitor image or from a tracing made of the image. PMID- 11900770 TI - Sexually conditioned incentives: attenuation of motivational impact during dopamine receptor antagonism. AB - The motivational impact of sexually conditioned incentives was examined in two experiments. In Experiment 1, male Long-Evans rats copulated to ejaculation in the presence of one of two scents (orange or almond extract) on five separate occasions. On alternating days, subjects spent an equal amount of time in social isolation with the opposing scent. Following the 10-day conditioning regimen, subjects ran more rapidly down an operant runway toward a goalbox containing the sex-paired scent (CS+) compared to trials on which the isolation-paired scent (CS ) or no scent was provided. In Experiment 2, comparably conditioned male rats were first given a baseline runway trial with an unscented goalbox. The following day, subjects were pretreated with one of four doses of haloperidol (0.0, 0.075, 0.15, or 0.30 mg/kg i.p.) 45 min prior to being tested in the runway for their motivation to approach either the CS+ or CS- scents. Control subjects given vehicle injections performed comparably to subjects from Experiment 1, taking significantly less time to approach the CS+ than an unscented goalbox. This decrease in run latency was not observed in subjects within the 0.075 and 0.15 mg/kg haloperidol groups. Subjects in the 0.30 mg/kg haloperidol groups took significantly more time to approach both the CS+ and CS- compared to their baseline run times. These data reveal that an olfactory cue associated with sexual reward becomes a conditioned incentive capable of eliciting approach behavior, and that dopamine receptor antagonism (at moderate but not high doses) selectively attenuates this cue-induced motivation. PMID- 11900772 TI - Stimulus generalization by fenfluramine in a quipazine-ketanserin drug discrimination is not dependent on indirect serotonin release. AB - The purpose of this study was to determine if animals trained to discriminate a serotonin2A (5-HT2A) receptor agonist from a 5-HT2A receptor antagonist would also be sensitive to alterations in serotonin neurotransmission brought about by 5-HT reuptake inhibitors and releasers. Previous work from our laboratory has shown that the quipazine-ketanserin discrimination is mediated solely by the 5 HT2A receptor, thus providing a behavioral continuum of 5-HT2A receptor function. Rats were trained to discriminate quipazine (0.35 mg/kg) from ketanserin (1.0 mg/kg) on a variable interval-30 schedule of reinforcement. Following acquisition, substitution tests were conducted with the training drug, quipazine, and agents that have been shown to alter the synaptic levels of 5-HT, including fenfluramine, norfenfluramine, 5-methoxy-6-methyl-2-aminoindan (MMAI) and fluoxetine. All compounds substituted, except fluoxetine. Antagonist tests with mianserin and MDL 100,907 indicated that fenfluramine's and MMAI's substitution for quipazine was mediated by the 5-HT2A receptor. Animals were pretreated with PCPA to determine whether 5-HT release or direct agonism mediated the discriminative stimulus effects of fenfluramine and MMAI. PCPA blocked the substitution of MMAI but not of fenfluramine for quipazine. Analysis of 3H-IP formation in cells showed that norfenfluramine dose-dependently stimulated phosphoinositide hydrolysis to levels similar to that of serotonin and quipazine. These results indicate that fenfluramine's substitution for quipazine in rats trained on a quipazine-ketanserin discrimination are due to direct agonism at the 5-HT2A receptor likely mediated by norfenfluramine, an active metabolite. PMID- 11900773 TI - The aversive properties of acute morphine dependence persist 48 h after a single exposure to morphine: evaluation by taste and place conditioning. AB - The aversive properties of naloxone-precipitated morphine withdrawal from acutely administered morphine were assessed following a single conditioning trial using both the place conditioning and the taste conditioning paradigm. In both paradigms, the aversive properties of naloxone-precipitated morphine withdrawal were evident up to 48 h after a single injection of morphine. In neither paradigm did naloxone treatment alone produce an aversion after a single conditioning trial. These results suggest that a single morphine exposure produces long lasting effects that persist at least 48 h beyond the agonist effects of the opiate. PMID- 11900774 TI - Role of Ca2+ channels on the hypothermic response produced by activation of kappa opioid receptors. AB - The effect of nimodipine (NIM) and lercanidipine (LER) 1,4-dihydropyridine (DHP) calcium channel blockers (CCBs) on the hypothermic response of selective kappa opioid receptor agonists U50,488H (U50), PD117,302 (PD) and U69,593 (U69) was determined in rats by recording colonic temperature using digital telethermometer. Intraperitoneal (i.p.) injections of U50 (7.5, 15, 22.5 and 40 mg/kg), PD (7.5, 15 and 22.5 mg/kg) and U69 (5 and 20 mg/kg) produced a dose dependent hypothermic response. However, higher doses of U50 (60 and 80 mg/kg) produced hypothermia, which is less when compared to that produced by 22.5-mg/kg dose of U50. NIM (1 mg/kg i.p.; 15 min prior) and LER (0.3 mg/kg i.p.; 15 min prior) did not produce any change in basal colonic temperature. Treatment of NIM and LER potentiated the U50 (7.5, 15, 22.5 and 40 mg/kg)-induced hypothermic effect. NIM did not potentiate hypothermia produced by U50 (60 mg/kg). On the other hand, PD (7.5, 15 and 22.5 mg/kg)- and U69 (5 and 20 mg/kg)-induced hypothermia was unaffected by the pretreatment of either NIM or LER. This differential modulation of kappa-opioid agonist-induced hypothermia by CCBs suggest that there may be two mechanisms, Ca(2+)-sensitive and Ca(2+) insensitive, involved in kappa-opioid agonist-induced hypothermic response. PMID- 11900775 TI - Prior experience with wheel running produces cross-tolerance to the rewarding effect of morphine. AB - The rewarding effect of wheel running is hypothesized to be mediated by endogenous opioids. Thus, prior experience with wheel running might be expected to affect the reward value of an opiate drug like morphine. In three similar experiments to test this idea, 10 rats (wheel-morphine group) were confined in running wheels for 2 h on each of eight consecutive days during the first phase; the 10 in the cage-morphine group were confined in small metal cages. Then, in the second phase, a distinctive place was paired with morphine (1 mg/kg) on three occasions to produce conditioned place preference (CPP). In all experiments, CPP occurred in the cage-morphine group, but not in the wheel-morphine group, implying that prior wheel running resulted in cross-tolerance to the rewarding effect of morphine. This finding supports the idea that the rewarding effect of wheel running is mediated by endogenous opioids. PMID- 11900776 TI - Context-specific tolerance to the ataxic effects of alcohol. AB - Tolerance to alcohol and many other drugs can become conditioned to specific contextual cues present at the time of drug administration. Context-specific tolerance occurs to a variety of alcohol's effects, including changes in hormone levels, body temperature and locomotor activity. The present study investigated whether context-specific tolerance can occur to the ataxic effects of alcohol. Baseline levels of motor coordination were assessed using a tilting plane apparatus. During a 7-day tolerance acquisition phase, subjects received an injection of either alcohol (1.5 g/kg i.p.) or saline (15 ml/kg i.p.) in a novel testing room and were then placed in the tilting plane apparatus for a period of 20 min. Approximately 5 h after the first injection, subjects received a second injection in the colony room and were then placed in their home cages. One group of subjects, the paired group, received alcohol in the testing room and saline in the colony room. An unpaired group received saline in the testing room and alcohol in the colony room. A no alcohol control group received saline in both environments. Following the tolerance acquisition phase, all subjects were injected with alcohol (1.5 g/kg i.p.) and tested for ataxia in the tilting plane apparatus. Subjects in the paired group were less ataxic than subjects in the control group during all four testing blocks following alcohol administration. In contrast, subjects in the unpaired group were less ataxic than the control subjects only during the 15-min testing block. Relative to baseline scores, the paired group exhibited deficits only during the 5- and 10-min testing blocks, while subjects in the unpaired and control groups exhibited deficits during all four testing blocks. These data strongly suggest that tolerance to the ataxic effects of alcohol can become conditioned to contextual cues present at the time of alcohol administration. PMID- 11900777 TI - Effects of GABAergic agents on anesthesia induced by halothane, isoflurane, and thiamylal in mice. AB - The effects of gamma-aminobutyric acid (GABA) receptor modulators and GABA uptake inhibitors on volatile and intravenous anesthetic-induced anesthesia were examined in male ICR mice, as assessed by the loss of righting reflex (LORR). The GABA uptake inhibitors, NO-711 and SKF89976A, which are permeable to the blood brain barrier (BBB), but not nipecotic acid or guvacine, which poorly permeate BBB, shortened the onset of LORR but did not affect the duration of LORR induced by 1.5% halothane and 2% isoflurane. NO-711 and SKF89976A shortened the onset of and prolonged the duration of LORR induced by thiamylal (45 mg/kg i.p.). The GABA mimetics, muscimol and diazepam, shortened the onset of and prolonged the duration of LORR induced by halothane, isoflurane, and thiamylal. On the other hand, picrotoxin, a GABAA receptor antagonist, prolonged the onset of LORR induced by all anesthetics tested. Another GABAA receptor antagonist, bicuculline, prolonged the onset of LORR induced by halothane, but not by isoflurane or thiamylal. Both antagonists failed to affect the duration of LORR induced by halothane, isoflurane, or thiamylal. Baclofen, a GABAB receptor agonist, enhanced both volatile anesthetics- and thiamylal-induced anesthesia. These results suggest that anesthesia induced by volatile and intravenous anesthetics might be correlated with the modification of the pre- and/or postsynaptic GABAergic activities. PMID- 11900778 TI - Increased drinking in mutant mice with truncated M5 muscarinic receptor genes. AB - The rarest and least understood of the muscarinic receptors is the M5 subtype. Recombinant methods were used to create mutant mice with a deletion in the third intracellular loop of the M5 receptor gene. Salivation induced by the nonselective muscarinic agonist pilocarpine (1 mg/kg s.c.) was reduced in homozygous mutants from 15 to 60 min after injection as compared with wild-type mice. After 18-h food and water deprivation, drinking was increased in homozygous mutants, but feeding was not increased. The mutant and wild-type mice had similar responses in tests of open-field exploration, seizures induced by pilocarpine (300 mg/kg) or hypothermia induced by pilocarpine (1-3 mg/kg). These results indicate that M5 muscarinic receptors are important for fluid intake and suggest that M5 receptors are involved in slow secretory processes. PMID- 11900779 TI - A simple procedure for assessing ataxia in rats: effects of phencyclidine. AB - The present study describes an objective, cost- and time-efficient procedure for characterizing the ataxic effects of psychoactive drugs. Male Sprague-Dawley rats were administered an intraperitoneal injection of either saline or one of three doses (1, 5 or 10 mg/kg) of phencyclidine (PCP) 15 min prior to being placed into an empty standard operant conditioning chamber (all manipulanda were removed). The floor of the test apparatus consisted of parallel rows of metal rods spaced approximately 1.5 cm apart. During a 5-min test, a single observer counted the frequency with which each animal's paws (front or back) slipped between the rows of bars that constituted the cage floor. The data demonstrated that while saline animals exhibited no instability in their ambulation, PCP-treated animals demonstrated a highly reliable dose-dependent increase in the number of "paw slips" in a single trial. Since animals are known to develop tolerance to the ataxic response to PCP, the validity of the test as a measure of drug-induced ataxia was examined in a separate group of animals treated with the middle (5 mg/kg) dose every other day over the course of a 9-day period (i.e., resulting in five injection trials). In this experiment, each subsequent test produced a reliable reduction in the magnitude of the ataxic response, and by the fifth drug challenge, the PCP animals were performing at near-control levels. These results suggest that the "paw slip test" can serve as a simple, reliable, objective and valid measure of drug-induced ataxia. The relevance of the ataxia data for interpreting the locomotor response of animals treated with PCP is also discussed. PMID- 11900780 TI - Behavioral effects of novel enterosorbent Noolit on mice with mixed depression/anxiety-like state. AB - The aim of this work was to examine the behavioral effects of a novel lithium based enterosorbent, Noolit (665 mg/kg), on male mice with mixed depression/anxiety-like state evoked by exposure to repeated social defeats in daily agonistic confrontations. The lithium component allows Noolit to be used as a psychotropic drug. Two experiments are described, in which the therapeutic and preventative effects of chronic Noolit treatment were examined. Response to Noolit was assessed in the plus maze, open field, partition test, and Porsolt's test. In both experiments, Noolit produced obvious anxiolytic and antidepressant effects. Treatment with Noolit fully restored some behavioral parameters in the plus maze and open field in depressed mice and prevented depression that would otherwise have developed. It has been suggested that enterosorbent Noolit can be a potent drug for the treatment of mixed anxiety/depression pathologies and for prevention of mood disorders. PMID- 11900781 TI - Characterization of the hypothermic component of LPS-induced dual thermoregulatory response in rats. AB - We have previously shown that Escherichia coli O111:B4 serotype lipopolysaccharide (LPS) produced a dual change in rectal temperature (Tb), in which hypothermia preceded fever at subthermoneutral ambient temperature (Tamb; 24-26 degrees C) in rats. In this study, the characteristics of the initial hypothermic response were evaluated. Hypothermia was significant when LPS (50 microg/kg, i.p.) was injected at thermoneutral Tamb (30 degrees C). There was no heat loss through tail skin during hypothermia. The open field activity of the rats did not change during this period. However, serum levels of tumor necrosis factor-alpha (TNF-alpha) elevated at the beginning of the hypothermia, whereas serum levels of interleukin (IL)-1beta and interferon (IFN)-gamma remained unchanged. A nonselective cyclooxygenase inhibitor (indomethacin, 5 mg/kg, s.c.) inhibited hypothermia and serum TNF-alpha elevation, which resulted in an acceleration of the subsequent pyrogenic response. Moreover, a nonselective inhibitor of nitric oxide synthase (nitro L-arginine methyl ester (L-NAME), 10 mg/kg, s.c.) not only abolished fever but also prolonged the initial hypothermic response. These data suggest that the hypothermic component of low dose LPS induced dual response is a regulated decrease in Tb. The data also suggest that hypothermia and fever may occur independently as two different thermoregulatory strategies against immune challenge in rats. PMID- 11900782 TI - Assessing the stimulant effects of alcohol in humans. AB - The stimulant effects of alcohol were assessed in humans. Twenty social drinkers were tested in dyads in the laboratory on three separate occasions, held 7 days apart. For their first session, one-third of the group consumed a dose of alcohol that was calculated to reach a target peak blood alcohol concentration (BAC) of 0.05 g/dl, one-third of the group consumed placebo-alcohol, and one-third consumed diet Sprite. For alcohol and placebo-alcohol conditions, subjects were told that they may or may not be given alcohol. For the soda condition, subjects were told they were consuming soda. Subjective stimulation, activity levels, and speech production were assessed over a 15-min period after beverage consumption (posttreatment) and compared to measurements taken prior to beverage consumption (baseline). Scores on the stimulant subscale of the Biphasic Alcohol Effects Scale (BAES) were significantly greater for the alcohol condition relative to the soda condition. There was also a trend for stimulant scores to be greater for the alcohol condition relative to the placebo-alcohol condition. Activity levels were significantly greater for the alcohol condition compared to either the placebo alcohol or soda conditions. There were no group differences found for speech production. Subjective stimulant score and activity levels were not correlated. Peak BAC obtained in subjects who consumed alcohol was not correlated with either subjective stimulant scores or activity levels. Activity levels may provide a useful behavioral assay for assessing the stimulant effects of alcohol in humans. PMID- 11900784 TI - Differential effects of prenatal morphine exposure on analgesia produced by vaginocervical stimulation or systemic morphine administration in adult rats. AB - The present study investigated the effects of prenatal morphine exposure on the magnitude of analgesia produced by vaginocervical stimulation (VS) or systemic morphine injection in adult rats. In Experiment 1, an acute subcutaneous morphine (1 mg/kg) injection induced a 124% greater increase in tail-flick latency (TFL) in adult rats exposed prenatally to saline than to morphine. By contrast, in Experiment 2, VS induced a 196% greater increase in TFL in adult rats exposed prenatally to morphine than to saline. Female rats exposed prenatally to morphine also had a greater VS-produced increase in vocalization threshold (VOC-T) to tail shock than those exposed prenatally to saline. Thus, the present study demonstrates that prenatal morphine exposure exerts diametrically opposite effects on analgesia that is produced in adulthood by morphine or VS, attenuating the former while potentiating the latter. These findings provide evidence that the mechanisms underlying the two types of analgesia differ fundamentally. PMID- 11900785 TI - 8-OHDPAT effects upon cocaine unconditioned and conditioned behaviors: a role for drug stimulus effects. AB - The effects of the 5-HT1A agonist, (+/-)-8-hydroxy-dipropylaminotetralin (8 OHDPAT) upon the unconditioned and conditioned behavior induced by cocaine were assessed in rats. Separate groups (n=7) received saline, cocaine (10 mg/kg), 8 OHDPAT (0.2 mg/kg), or 8-OHDPAT (0.2 mg/kg) plus cocaine (10 mg/kg) for eight treatment sessions (two per week) in which the rats were tested for 20 min in an open-field. On the eighth treatment session, cocaine enhanced locomotion and rearing but decreased grooming. 8-OHDPAT also decreased grooming and, when given in combination with cocaine, enhanced locomotion but attenuated cocaine-induced rearing. The two 8-OHDPAT groups differed substantially from each other and from the cocaine group in terms of locomotion during the drug treatment phase. Subsequently, all groups received a series of conditioning tests in which they received saline, 0.1, 0.2, or 0.4 mg/kg 8-OHDPAT prior to testing. Groups which had received either 8-OHDPAT or cocaine prior to the conditioning tests exhibited equivalent conditioned effects on the saline conditioning test. When conditioning tests were conducted with 8-OHDPAT, however, only the group which had previously received the combined 0.2 mg/kg 8-OHDPAT plus cocaine treatment exhibited a conditioned response and this effect only occurred at the 0.2 8-OHDPAT dose level. These observations indicate the important influence of the stimulus properties of drugs for the study of drug conditioning and for understanding drug interactions with cocaine. PMID- 11900783 TI - Chronic opiate exposure in the male rat adversely affects fertility. AB - We examined whether morphine administration to adult male rats adversely affected pregnancy outcome after mating with drug-naive females and at what point in the complex series of steps leading to viable offspring it exerted its actions. The results indicate that chronic paternal morphine exposure markedly influenced fertility measures in a number of important ways. There was a pronounced increase in pseudopregnancies in females mated with males treated chronically with morphine (40%) when compared to controls (<6%), indicating that vaginal penetration occurred, but successful impregnation failed; only 33% of matings between drug-naive females and morphine-treated males resulted in pregnancies, as compared to 74.5% in controls. In addition, there were fewer implantation sites in gravid females mated with morphine-treated males than in controls. Taken together, these observations suggest that morphine-exposed male rats were apparently able to copulate, but there was a failure in successful impregnation of the females. These findings suggest a primary defect in either the quality of male sexual behavior or a complete failure of the fertilization or conception processes in females mated with morphine-exposed males. This potentially important effect of paternal morphine administration on conception and/or preimplementation loss of embryos has not been previously noted and deserves more systematic study. PMID- 11900786 TI - GABAA-benzodiazepine receptor complex ligands and stress-induced hyperthermia in singly housed mice. AB - Stress-induced hyperthermia (SIH) in singly housed mice, in which the rectal temperature of a mouse is measured twice with a 10-min interval, enables to study the effects of a drug on the basal (T1) and on the stress-enhanced temperature (T2), 10 min later, using the rectal procedure as stressor. SIH (T2-T1) reflects a stress-induced phenomenon sensitive to stress- or anxiety-modifying effects of drugs. Several benzodiazepine agonists (diazepam, chlordiazepoxide, oxazepam and alprazolam) dose-dependently antagonized SIH either in NMRI mice from two different breeders or in BALB/c mice. No major differences in the sensitivity for any of the drugs tested were found between strains or between substrains from different breeders. The selective BZ1 receptor agonists alpidem and zolpidem only at relatively high doses antagonized SIH, whereas flumazenil, FG7142, pentylenetetrazol and phenobarbital did not affect SIH. Alcohol antagonized SIH, and the effects of diazepam could be antagonized by flumazenil. The findings that full BZ receptor agonists have anxiolytic-like effects in the singly housed SIH paradigm are comparable to those previously found in the group-housed version. The singly housed SIH is proposed as a simple and reliable screen for detecting anxiety-like properties of drugs that is valid in every mouse strain tested so far. PMID- 11900788 TI - Nicotine potentiation of morphine-induced catalepsy in mice. AB - In the present study, effects of nicotine on catalepsy induced by morphine in mice have been investigated. Morphine but not nicotine induced a dose-dependent catalepsy. The response of morphine was potentiated by nicotine. Intraperitoneal administration of atropine, naloxone, mecamylamine, and hexamethonium to mice reduced catalepsy induced by a combination of morphine with nicotine. Intracerebroventricular injection of atropine, hexamethonium, and naloxone also decreased catalepsy induced by morphine plus nicotine. Intraperitoneal administration of atropine, but not intraperitoneal or intracerebroventricular injection of hexamethonium, decreased the effect of a single dose of morphine. It was concluded that morphine catalepsy can be elicited by opioid and cholinergic receptors, and the potentiation of morphine induced by nicotine may also be mediated through cholinergic receptor mechanisms. PMID- 11900787 TI - Asymmetric calmodulin distribution in the hypothalamus: role of sexual differentiation in the rat. AB - The Ca2+/calmodulin (CaM) system plays important roles both in hypothalamic sexual differentiation and in the progesterone-induced facilitation of lordosis behavior in the adult rat. We recently showed sex-dependent differences in rat hypothalamic CaM levels, both in newborn and in adult animals. Here, we evaluated the presence of left-right hypothalamic asymmetries in CaM concentration in male and female rats, as well as the changes induced on these parameters by neonatal (1 h after birth) subcutaneous administration of tamoxifen (200 microg/rat) or testosterone (30 microg/rat). CaM was measured by RIA in each half of the hypothalamus (at 2, 6, 12, and 24 h and at 90 days after birth) in both control and treated animals. In untreated young rats (2-24 h after birth), CaM concentration was significantly higher in the right half of the hypothalamus of males, whereas in females, it was higher in the hypothalamic left half. Treatment of females with testosterone or tamoxifen to males, consistently reversed these results. In the hypothalamus of treated animals, we found higher CaM levels in the left half of males, as well as in the right half of females. In control adult females, CaM concentration was also higher in the left half of the hypothalamus, as it was in the right half of adult males. However, this asymmetry was lost after neonatal hormone manipulation. These results reinforce the role of CaM in the development of sex-related hypothalamic functions. PMID- 11900789 TI - Effects of adrenoceptor agonists and antagonists on morphine-induced Straub tail in mice. AB - Straub-tail behavior was induced by subcutaneous injection of different doses (10 60 mg/kg) of morphine to mice. The maximum response was obtained with 20-40 mg/kg of the drug. The response induced by morphine (40 mg/kg) was decreased by intraperitoneal administration of different doses of clonidine (0.05-0.1 mg/kg). Pretreatment of animals with yohimbine (1-4 mg/kg i.p.) reversed the inhibitory action of clonidine. Yohimbine did not elicit any response by itself. Administration of prazosin (0.25, 0.5, and 1 mg/kg) reduced the morphine response. The combination of prazosin with yohimbine (1 mg/kg), but not with clonidine (0.05 mg/kg), caused a potentiated inhibition of the morphine effect. Phenylephrine (2-6 mg/kg i.p.) did not elicit any effect by itself and also did not alter the response induced by morphine or morphine plus clonidine. Dobutamine (2.5-10 mg/kg i.p.), atenolol (2.5-10 mg/kg i.p.), salbutamol (2.5-10 mg/kg i.p.), and propranolol (2.5-10 mg/kg i.p.) did not alter morphine-induced Straub tail behavior in mice. In conclusion, activation of alpha2-adrenergic pathways contributes to morphine-induced Straub tail, while alpha1- and beta2-adrenergic may not be involved in this phenomenon. PMID- 11900790 TI - Basal forebrain infusions impair delayed-non-match-to-sample radial arm maze performance. AB - Direct site infusion of drugs into the brain is a powerful tool for examining the function of specific brain regions. While comparing the effects of various drugs injected into distinct regions of the basal forebrain on cognitive and motor endpoints, we observed that the ventral pallidum (VP) appeared to be sensitive to vehicle control infusions when cognitive indices were measured. To characterize this initial observation, the present study examined the effects of vehicle infusions into the VP on performance of a delayed-non-match-to-sample (DNMTS) radial arm maze (RAM) task. A within-subjects design was used. Male Sprague Dawley rats were trained to perform this task with a 1-h delay imposed between the fourth and fifth arm selection. Following acquisition, animals were implanted with bilateral, indwelling cannulae positioned over the VP. Following surgery, maze performance was reestablished and rats were given one of five intra-VP treatments (two sham and three vehicle infusions) in counterbalanced order. Each rat received one treatment a week, on the third day of five consecutive testing days each week. Vehicle, but not sham, treatments produced deficits on the day of treatment and on two subsequent testing days. These findings demonstrate a persistent sensitivity of the VP to fluid perturbation and, when contrasted with the literature for other basal forebrain regions, it appears that this effect is unique. PMID- 11900791 TI - Elevated extracellular CRF levels in the bed nucleus of the stria terminalis during ethanol withdrawal and reduction by subsequent ethanol intake. AB - Corticotropin-releasing factor (CRF) is widely distributed throughout the brain and has been shown to mediate numerous endocrine and behavioral responses to stressors. During acute ethanol withdrawal, CRF release is increased in the central nucleus of the amygdala (CeA), and there is evidence to suggest that this activation of amygdala CRF systems may mediate the anxiogenic properties of the ethanol withdrawal syndrome. The present study was conducted to determine if another CRF-containing limbic structure, the bed nucleus of the stria terminalis (BNST), we would exhibit similar increases in CRF neurotransmission during ethanol withdrawal. Rats were administered an ethanol-containing (6.7% v/v) or control liquid diet for 2 weeks and subsequently implanted with microdialysis probes into the lateral BNST. A 50-75% increase in dialysate CRF levels was observed following removal of the ethanol-containing diet, while no changes were observed in control animals. When ethanol-withdrawn animals were given subsequent access to the ethanol-containing diet, dialysate CRF levels returned to basal levels. However, when ethanol-withdrawn animals were given subsequent access to the control diet, dialysate CRF levels increased further to 101% above basal levels. These data demonstrate that extracellular CRF levels are increased in the BNST during ethanol withdrawal, and that these increases are reduced by subsequent ethanol intake. PMID- 11900792 TI - Gonadal hormone-induced changes in adult male and female rats exposed to early postnatal handling are not altered by prenatal morphine exposure. AB - The purpose of the present study was to investigate the long-term effects of early postnatal handling in several gonadal hormone conditions in adult male and female rats exposed prenatally to morphine or saline. An open-field apparatus was used to test locomotor activities such as line crossing, rearing, grooming, and anxiety-like behaviors such as visiting squares alongside the walls of the open field and boli dropping. Postnatal handling increased locomotor activities in gonadally intact males and in all groups of hormone-manipulated females, but did not change them in gonadectomized (GNX) males. Additionally, there was a decrease in anxiety-like behavior in ovariectomized (OVX) females after estradiol benzoate (EB) or EB and progesterone (P) replacement due to handling. Handling did not affect anxiety-like behaviors in OVX females or in GNX or gonadally intact males. Prenatal morphine exposure did not alter any open-field measures in handled or nonhandled animals when compared to saline controls. Thus, the present study demonstrates that early postnatal handling induces long-lasting changes in locomotor and anxiety-like behaviors of adult male and female rats regardless of their prenatal exposure to morphine. These changes are gonadal hormone specific. PMID- 11900793 TI - Role of nitric oxide and beta-adrenoceptors of the central nervous system on the salivary flow induced by pilocarpine injection into the lateral ventricle. AB - Our studies have focused on the effect of L-NG-nitroarginine methyl ester (L NAME), an inhibitor of nitric oxide synthase (NOS), and L-arginine, the substrate of NOS, on salivary secretion induced by the administration of pilocarpine into the lateral cerebral ventricle (LV) of rats. The present study has also investigated the role of the beta-adrenergic agonists and antagonist injected into LV on the salivary secretion elicited by the injection of pilocarpine into LV. Male Holtzman rats with a stainless-steel cannula implanted into the LV were used. The amount of salivary secretion was studied over a 7-min period after injection of pilocarpine, isoproterenol, propranolol, salbutamol, salmeterol, L NAME and L-arginine. The injection of pilocarpine (10, 20, 40, 80 and 160 microg/microl) into LV produced a dose-dependent increase in salivary secretion. The injection of L-NAME (40 microg/microl) into LV alone produced an increase in salivary secretion. The injection of L-NAME into LV previous to the injection of pilocarpine produced an increase in salivary secretion. L-Arginine (30 microg/microl) injected alone into LV produced no change in salivary secretion. L Arginine injected into LV attenuated pilocarpine-induced salivary secretion. The isoproterenol (40 nmol/microl) injected into LV increased the salivary secretion. When injected previous to pilocarpine at a dose of 20 and 40 microg/microl, isoproterenol produced an additive effect on pilocarpine-induced salivary secretion. The 40-nmol/microl dose of propranolol injected alone or previous to pilocarpine into LV attenuated the pilocarpine-induced salivary secretion. The injection of salbutamol (40 nmol/microl), a specific beta-2 agonist, injected alone into LV produced no change in salivary secretion and when injected previous to pilocarpine produced an increase in salivary secretion. The 40-nmol/microl dose of salmeterol, a long-acting beta-2 agonist, injected into LV alone or previous to pilocarpine produced no change in salivary secretion. The results have shown that central injections of L-NAME and L-arginine interfere with the salivary secretion, which implies that might participate in pilocarpine-induced salivary secretion. The interaction between cholinergic and beta-adrenergic receptors of the central nervous system (CNS) for the control of salivary secretion can also be postulated. PMID- 11900795 TI - Two related forms of memory in the crab Chasmagnathus are differentially affected by NMDA receptor antagonists. AB - A visual danger stimulus (VDS) elicits an escape response in the crab Chasmagnathus that declines after a few iterative presentations. Long-lasting retention of such decrement, termed context-signal memory (CSM), is mediated by an association between danger stimulus and environmental cues, cycloheximide sensitive, correlated with PKA activity and NFkappa-B activation, positively modulated by angiotensins, and selectively regulated by a muscarinic-cholinergic mechanism. The present research was aimed at studying the possible involvement of NMDA-like receptors in CSM, given the role attributed to these receptors in vertebrate memory and their occurrence in invertebrates including crustaceans. Vertebrate antagonists (+/-)-2-amino-5-phosphonopentanoic acid (AP5) and (+)-5 methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801) were used. Memory retention impairment was shown with MK-801 10(-3) M (1 microg/g) injected immediately before training or after training, or delayed 1 or 4 h, but not 6 h, posttraining. An AP5 10(-3) M dose (0.6 microg/g) impairs retention when given before but not after training. Neither antagonist produced retrieval deficit. A memory process similar to CSM but nonassociative in nature and induced by massed training (termed signal memory, SM), proved entirely insensitive to AP5 or MK 801, confirming the view that distinct mechanisms subserve these different types of memory in the crab. PMID- 11900794 TI - Effects of frequent marijuana use on memory-related regional cerebral blood flow. AB - It is uncertain whether frequent marijuana use adversely affects human brain function. Using positron emission tomography (PET), memory-related regional cerebral blood flow was compared in frequent marijuana users and nonusing control subjects after 26+ h of monitored abstention. Memory-related blood flow in marijuana users, relative to control subjects, showed decreases in prefrontal cortex, increases in memory-relevant regions of cerebellum, and altered lateralization in hippocampus. Marijuana users differed most in brain activity related to episodic memory encoding. In learning a word list to criterion over multiple trials, marijuana users, relative to control subjects, required means of 2.7 more presentations during initial learning and 3.1 more presentations during subsequent relearning. In single-trial recall, marijuana users appeared to rely more on short-term memory, recalling 23% more than control subjects from the end of a list, but 19% less from the middle. These findings indicate altered memory related brain function in marijuana users. PMID- 11900796 TI - The involvement of nitric oxide in the antinociception induced by cyclosporin A in mice. AB - Cyclosporin A (CsA) and other immunophilin-binding agents are known to inactivate neuronal nitric oxide synthase (nNOS). Nitric oxide (NO) is involved in the nociception at the spinal level. We evaluated the effect of acute intraperitoneal (i.p.) administration of CsA on the tail-flick response in mice and the involvement of NO and opioid receptors in this effect. CsA (5, 10, 20 and 50 mg/kg i.p.) induced a significant increase in tail-flick response. Nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine (LNNA; 10, 40 and 80 mg/kg i.p.) significantly potentiated the CsA-induced (5 mg/kg) increase in tail-flick latency (TFL). While NOS substrate L-arginine (100, 200, 400 mg/kg i.p.) inhibited the CsA-induced (20 mg/kg) antinociception completely and in a dose dependent manner. Concomitant administration of L-NNA and L-arginine blocked the inhibition exerted by the latter on the CsA-induced antinociception. The opioid receptor antagonist naloxone (4 mg/kg i.p.) did not alter the CsA effect. These results indicate that acute administration of CsA induces an antinociceptive effect that involves the L-arginine-NO pathway but is not mediated by opioid receptors. PMID- 11900797 TI - Mu-opioid receptor down-regulation and tolerance are not equally dependent upon G protein signaling. AB - In the present study, the contribution of pertussis toxin (PTX)-sensitive G(i/o) proteins to opioid tolerance and mu-opioid receptor down-regulation in the mouse were examined. Mice were injected once intracerebroventricularly and intrathecally with PTX (0.1 microg/site). Controls were treated with saline. On the 10th day following PTX treatment, continuous subcutaneous infusion of etorphine (150 or 200 microg/kg/day) or morphine (40 mg/kg/day+25 mg slow-release pellet) was begun. Control mice were implanted with inert placebo pellets. Pumps and pellets were removed 3 days later, and mice were tested for morphine analgesia or mu-opioid receptor density was determined in the whole brain, spinal cord, and midbrain. Both infusion doses of etorphine produced significant tolerance (ED50 shift=approximately 4-6-fold) and down-regulation of mu-opioid receptors (approximately 20-35%). Morphine treatment also produced significant tolerance (ED50 shift= approximately 5-8-fold), but no mu-opioid receptor down regulation. PTX dramatically reduced the acute potency of morphine and blocked the further development of tolerance by both etorphine and morphine treatments. However, PTX had no effect on etorphine-induced mu-opioid receptor down regulation in brain, cord, or midbrain. These results suggest that PTX-sensitive G-proteins have a minimal role in agonist-induced mu-opioid receptor density regulation in vivo, but are critical in mediating acute and chronic functional effects of opioids such as analgesia and tolerance. PMID- 11900798 TI - Continuous nicotine infusion reduces nicotine self-administration in rats with 23 h/day access to nicotine. AB - The effects of continuous nicotine infusion on nicotine self-administration (NSA) were studied in rats as a model of nicotine replacement therapy (NRT) in humans. A NSA model in which rats had 23-h/day access to nicotine was used to approximate nicotine access conditions in cigarette smokers. In order to estimate serum nicotine concentrations associated with NSA, arterial and venous serum nicotine concentrations were measured during a simulation of NSA. Nicotine was noncontingently administered as 30 doses/12 h of 0.03 mg/kg/i.n.f. or 60 doses/12 h of 0.01 mg/kg/i.n.f. daily. Venous serum nicotine concentrations were measured after the first nicotine dose of the day, and arterial and venous concentrations were measured after doses in the middle of the day. The range of mean concentrations measured was similar to those reported in cigarette smokers (venous concentrations 6-59 ng/ml, arterial concentrations 42-96 ng/ml). The effects of continuous nicotine infusion on NSA were studied by noncontingently administering nicotine at various rates via osmotic pump to animals self administering nicotine (0.01 or 0.03 mg/kg/i.n.f.) during 23-h/day sessions. Continuous nicotine infusion at all infusion rates substantially suppressed NSA, but suppression was rate-related only for the 0.01-mg/kg/inf NSA unit dose. Nicotine infusion rates producing venous serum nicotine concentrations equaling or exceeding the peak venous levels associated with simulated NSA were more effective than lower infusion rates only at the lower NSA unit dose. The highest nicotine infusion rate had no sustained effect on food-maintained responding, demonstrating its specificity for suppression of NSA. These data provide a model for studying NRT in the rat. PMID- 11900799 TI - Effect of NMDA antagonists, an NMDA agonist, and serotonin depletion on acute tolerance to ethanol. AB - The effect of N-methyl-D-aspartate (NMDA) antagonists [dizocilpine, (+)MK-801, and ketamine], an NMDA agonist (D-cycloserine) and of brain serotonin (5-HT) depletion with p-chlorophenylalanine (p-CPA) on acute tolerance to ethanol was examined, using the tolerance model proposed by Radlow [Psychopharmacology 114 (1994) 1-8] and Martin and Moss [Alcohol Clin Exp Res 17 (1993) 211-216]. This model is based on the concept of a linear increase of acute tolerance with time; the rate of acute tolerance development is the slope of the output function that relates blood alcohol concentrations (BACs) and intoxication. Pretreatment with NMDA antagonists inhibited the development of acute tolerance to ethanol, whereas pretreatment with D-cycloserine enhanced it. Depletion of 5-HT by p-CPA also blocked acute tolerance to ethanol. These results on acute tolerance are similar to those previously found on rapid and chronic tolerance to ethanol. PMID- 11900800 TI - Effect of PMA optical isomers and 4-MTA in PMMA-trained rats. AB - 1-(4-Methoxyphenyl)-2-aminopropane (PMA) and its sulfur analog, 1-(4 methylthiophenyl)-2-aminopropane (4-MTA), have been misrepresented as the controlled substance analog, N-methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane (MDMA; "Ecstasy"). Because MDMA has been shown to produce both amphetamine-like and N-methyl-1-(4-methoxyphenyl)-2-aminopropane (PMMA)-like stimulus effects in rats, we examined S(+)PMA, R(-)PMA and 4-MTA in rats trained to discriminate either PMMA (1.25 mg/kg) or (+)amphetamine (1.0 mg/kg) from saline vehicle. The sulfur analog of PMMA (i.e., 4-MTMA) was also examined. The PMMA stimulus generalized to R(-)PMA (ED50=0.4 mg/kg), whereas S(+)PMA produced a maximum of 72% PMMA-appropriate responding. 4-MTA (ED50=0.3 mg/kg) also substituted for PMMA, but 4-MTMA produced a maximum of only 36% PMMA-appropriate responding. None of the four agents substituted for (+)amphetamine. Hence, like MDMA, R(-)PMA and 4-MTA are capable of producing PMMA stimulus effects in rats, but unlike MDMA, neither agent substituted for (+)amphetamine. PMID- 11900801 TI - Effect of 1-(3,4-methylenedioxyphenyl)-2-aminopropane and its optical isomers in PMMA-trained rats. AB - 1-(3,4-Methylenedioxyphenyl)-2-aminopropane (MDA) is a drug of abuse that is known to produce stimulus effects similar to those of the stimulant phenylalkylamine (+)amphetamine and the hallucinogenic phenylalkylamine 1-(2,5 dimethoxy-4-methylphenyl)-2-aminopropane (DOM). Earlier, a working model was described to account for the stimulus effects produced by phenylalkylamines. Such agents can produce one or more of three distinct effects: an amphetamine effect, a DOM effect and a third effect that is typified by the agent N-methyl-1-(4 methoxyphenyl)-2-aminopropane (PMMA). Because MDA is known to produce two of the three effects, in the present investigation, we sought to determine if racemic MDA or either of its optical isomers could produce a PMMA-like effect in animals. Administration of S(+)MDA, R(-)MDA and (+/-)MDA to rats trained to discriminate 1.25 mg/kg of PMMA from saline vehicle under a VI 15-s schedule of reinforcement resulted in substitution in each case. (+/-)MDA and S(+)MDA were nearly equipotent and several fold more potent than R(-)MDA. The results are not only consistent with the proposed model but also identify (+/-)MDA as the first phenylalkylamine shown to produce all three types of stimulus effects (i.e., amphetamine-like, DOM-like and PMMA-like) in rats. PMID- 11900802 TI - Influence of estrogen in the acquisition of intravenously self-administered heroin in female rats. AB - Previous research indicates that female rats acquire cocaine and heroin self administration at a faster rate than male rats, and female rats with endogenous estrogen, or ovariectomized (OVX) rats with estrogen replacement acquire cocaine self-administration more rapidly than female rats with estrogen either surgically or chemically blocked. The purpose of this investigation was to extend the above findings to the acquisition of heroin (0.0075 mg/kg) self-administration in female rats. An automated autoshaping procedure was used to train rats to self administer heroin. Three groups of female rats were compared: (1) OVX+estradiol benzoate (OVX+EB), (2) OVX+vehicle (OVX+VEH), and (3) sham-operated+vehicle (SH+VEH). Results revealed that OVX+EB rats acquired heroin self-administration in significantly fewer days compared to OVX+VEH rats. Additionally, OVX+EB rats that met the acquisition criteria self-administered a significantly greater number of heroin infusions during the last 5 days of the acquisition period compared to OVX+VEH rats. These results indicate that OVX+EB rats initiate heroin use sooner than OVX+VEH rats and consume greater amounts of heroin during the last 5 days of acquisition compared to OVX+VEH female rats. PMID- 11900803 TI - Cholinergic blockade impairs performance in operant DNMTP in two inbred strains of mice. AB - Cholinergic blockade has been shown to impair performance in delayed nonmatching to position (DNMTP) paradigms in rats. In this study, a murine operant DNMTP task was used to assess the effects of cholinergic antagonism in two strains of mice (DBA/2 and C57BL/6) differing in spatial learning abilities. DNMTP was scheduled in operant chambers with retractable levers, where mice were trained until high levels of accuracy. Subsequently, proactive interference effects were assessed by manipulation of the intertrial interval (ITI), and animals were tested in this task under scopolamine (0.1-1.0 mg/kg) and mecamylamine (0.5-2.0 mg/kg) treatment. Data were analyzed according to the methods of signal detection theory. ITI manipulation decreased accuracy when the time between trials was reduced to 5 s. Cholinergic blockade failed to induce a pure mnemonic impairment but distinguishable effects of both receptor antagonists could be detected: scopolamine disrupted accuracy in a dose-dependent but delay-independent manner, whereas mecamylamine failed to impair accuracy, but decreased responsivity delay- and dose-dependently. Strains mainly differed in responsivity, with DBA/2 showing higher latencies to respond to the levers. These results are comparable to those obtained in rats. Thus, operant DNMTP can be applied to assess working memory in mice. PMID- 11900804 TI - A comparison of female and male rats' ETOH-induced ataxia and exploration following restraint or swim stress. AB - Animal models of stress reactivity are often employed in developing treatments for humans. Many studies use shock stress, and most use male rats. These experiments compare female and male rats exposed to either restraint stress (RS) or ambient-temperature swim stress (SS), using two durations of each stressor and naive controls. The ataxic effects of a 0.6 g/kg i.p. dose of ethanol (ETOH) were measured. Females exhibited less ataxia than males following ETOH administration. There were no significant effects of stress on ETOH-induced ataxia. Exploration was also measured in an open-field test (OFT) both pre- and poststress. In the prestress OFT, females were more active than males. For the no-stress groups and the shorter-duration stress groups, exploration decreased between the first and second OFTs. However, the groups exposed to the longer-duration stress did not show this expected decrease in exploration. A key finding of this research is that while sex differences may be present at baseline, the sexes may react similarly to stress. These data extend knowledge on sex differences in stress, alcohol reactivity and exploratory behavior. PMID- 11900805 TI - BTCP is a potent reinforcer in rats: comparison of behavior maintained on fixed- and progressive-ratio schedules. AB - N-[1-(2-benzo[b]thiophenyl)cyclohexyl]piperidine (BTCP) is a phencyclidine (PCP) derivative that acts as a potent dopamine (DA) reuptake inhibitor. Earlier studies have shown that BTCP can substitute for the reinforcing effects of cocaine. Therefore, the aim of the study was to further characterize the reinforcing effects of BTCP. The reinforcing actions of BTCP were compared to those of cocaine at equimolar concentrations in drug-naive rats. Two groups of animals were implanted with jugular catheters and trained to intravenously self administer BTCP or cocaine (0.25 mg/infusion) on a fixed-ratio five schedule (FR 5) of reinforcement. Both BTCP and cocaine produced comparable inverted U-shaped dose-effect curves on this schedule over doses of 0.03, 0.06, 0.125, and 0.25 mg/infusion. Two doses (0.125 and 0.25 mg/infusion) that produced reliable self administration in all the animals for cocaine and BTCP were then tested on a progressive-ratio schedule. At each dose, BTCP supported higher breaking points (BPs) than cocaine. The results demonstrate that rats readily acquire responding maintained by BTCP and suggest that BTCP may have greater reinforcing effects than cocaine at equimolar concentrations. PMID- 11900806 TI - Effects of nicotine on alcohol intake in a rat model of depression. AB - Clinical studies suggest that depression facilitates alcohol abuse. Depressed individuals also have increased rates of smoking, and it has been suggested that nicotine may improve depression. It is therefore possible that nicotine may reduce alcohol use in depression. To investigate this potential relationship, we evaluated alcohol intake in an animal model of depression, which consists of administering clomipramine (CLI), a preferential serotonin reuptake inhibitor, to neonatal rats. This pharmacological manipulation produces adult depression-like behaviors, such as reduced aggressiveness, decreased pleasure seeking, diminished sexual activity, increased locomotor activity and increased REM sleep. In this study, we found that CLI rats exhibited significantly higher locomotor activity, lower aggressiveness and higher alcohol intake than control rats. Chronic administration of a low dose of nicotine (0.25 mg/kg/day) or a sham operation did not modify these behaviors. However, chronic administration of nicotine at a higher dose (1.5 mg/kg/day) significantly increased aggressive behavior and reduced alcohol intake in CLI rats. The effect of nicotine on alcohol intake lasted at least 1 month after cessation of nicotine administration. These results indicate that nicotine reverted some depression signs and reduced alcohol self administration in the CLI model of depression. PMID- 11900807 TI - Prazosin inhibits spontaneous locomotor activity in diabetic mice. AB - We examined the effect of prazosin, a selective alpha1-adrenergic receptor antagonist, on the enhanced spontaneous locomotor activity in streptozotocin induced diabetic mice. Spontaneous locomotor activity in diabetic mice was significantly greater than that in nondiabetic mice. Pretreatment with either intracerebroventricular (5, 10 nmol) or intraperitoneal (0.5, 1.0 mg/kg) injection of prazosin dose-dependently reduced the spontaneous locomotor activity in diabetic mice, but not in nondiabetic mice. Furthermore, the enhanced dopamine turnover ratio in the limbic forebrain in diabetic mice was reduced to the same level as that in nondiabetic mice after the administration of prazosin. Thus, these results suggest that alpha1-adrenergic receptors might play an important role in the enhanced spontaneous locomotor activity in diabetic mice. Furthermore, alpha1-adrenoceptor antagonism might have an inhibitory effect on presynaptic dopaminergic neurotransmission in the limbic forebrain in diabetic mice. PMID- 11900808 TI - A novel behavioral model that discriminates between 5-HT2A and 5-HT2C receptor activation. AB - 2,5-Dimethoxy-4-iodoamphetamine (DOI), a serotonin (5-HT)2A/2C receptor agonist, elicits shaking behaviors in rodents, which have been reliably quantified as behavioral correlates of 5-HT2A receptor activation. Such studies are lacking in the rabbit. As part of our research examining the role of the 5-HT2 receptor in rabbits, we analyzed the behavioral effects of systemically administered DOI in rabbits. DOI (0.01-3 micromol/kg) or vehicle was injected, and two distinct behaviors, head bobs (vertical head movements) and body shakes (wet dog shakes), were counted for 90 min following the injection. DOI dose-dependently increased the number of head bobs and body shakes. The selective 5-HT2A receptor antagonist ketanserin (1-3 micromol/kg), 1 h before DOI (0.3 micromol/kg) challenge, significantly attenuated head bobs, but not body shakes. In contrast, the selective 5-HT2C receptor antagonists SDZ SER 082 (1-3 micromol/kg) and SB 206553 (1 micromol/kg) 30 min before challenge, significantly reduced body shakes but not head bobs produced by the same dose of DOI. This study establishes that, in rabbits, DOI mediates head bobs via 5-HT2A receptors and body shakes via 5-HT2C receptors. Thus, the rabbit provides a novel behavioral assay that discriminates between 5-HT2A and 5-HT2C receptor activation. PMID- 11900809 TI - Further characterization of the stimulus properties of 5,6,7,8-tetrahydro-1,3 dioxolo[4,5-g]isoquinoline. AB - This investigation is based on the premise that conformational restriction of abused phenylalkylamines in a tetrahydroisoquinoline conformation alters their pharmacology in such a manner that their original action is lost and that a new action emerges. TDIQ or 5,6,7,8-tetrahydro-1,3-dioxolo[4,5-g]isoquinoline, is a conformationally constrained phenylalkylamine that serves as a discriminative stimulus in animals. Although TDIQ bears structural resemblance to phenylalkylamine stimulants (e.g., amphetamine), hallucinogens (e.g., 1-(2,5 dimethoxy-4-methylphenyl)-2-aminopropane [DOM]), and designer drugs (e.g., N methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane [MDMA], N-methyl-1-(4 methoxyphenyl)-2-aminopropane [PMMA]), the TDIQ stimulus failed to generalize to (+)amphetamine or MDMA. In the present investigation, further evaluations were made of the stimulus nature of TDIQ. Specifically, the stimulus similarities of TDIQ, PMMA, and DOM were examined. In no case was stimulus generalization (substitution) observed. The results confirm that TDIQ produces stimulus effects distinct from those of the abovementioned phenylalkylamines. We also examined the structure-activity relationships of a series of TDIQ analogs, including several that might be viewed as conformationally restricted (CR) analogs of phenylalkylamine hallucinogens, stimulants, and designer drugs. These agents were examined in rats trained to discriminate either DOM (1.0 mg/kg), (+)amphetamine (1.0 mg/kg), MDMA (1.5 mg/kg), or TDIQ (5.0 mg/kg) from saline vehicle. Whereas we have demonstrated that none of these agents retains their respective phenylalkylamine stimulus actions, several of these agents were found to substitute for TDIQ. N-Methylation abolished TDIQ-stimulus action. These results, coupled with previous findings, imply that TDIQ derivatives represent a novel class of phenylalkylamines analogs with unique stimulus properties. Preliminary radioligand binding studies suggest that an alpha2-adrenergic mechanism might underlie the stimulus effects produced by TDIQ. PMID- 11900810 TI - The effect of cyanamide and 4-methylpyrazole on the ethanol-induced locomotor activity in mice. AB - To assess the role of cyanamide and 4-methylpyrazole (4-MP) in mediating ethanol induced locomotor activity in mice, they were pretreated with cyanamide (12.5, 25, or 50 g/kg) prior to one ethanol injection (2.4 g/kg) and showed significantly depressed locomotor activity compared with control groups. Cyanamide (25 mg/kg) also cancelled out the biphasic action of ethanol (0, 0.8, 1.6, 2.4, 3.2, or 4 g/kg) on locomotor activity. The action of cyanamide and 4-MP in combined administration was also tested. Our data show that pretreatment with 4-MP alone does not change the spontaneous or ethanol-induced locomotor activity. Conversely, when mice were pretreated with cyanamide and 4-MP, the depressive effect of cyanamide on the locomotor activity induced by ethanol disappeared, and the locomotor activity rose to levels similar to those of the control group, recovering the biphasic ethanol effect. These effects cannot be attributed to peripheral elevated blood acetaldehyde levels, as pretreatment with 4-MP prevents accumulation of acetaldehyde. These data might suggest some influence of brain catalase and aldehyde dehydrogenase (ALDH) on the effects of ethanol. PMID- 11900811 TI - Effects of imipramine and lithium on wet-dog shakes mediated by the 5-HT2A receptor in ACTH-treated rats. AB - We examined the influence of imipramine and lithium on wet-dog shakes induced by the (+/-)-DOI, 5-HT2A receptor agonist in adrenocorticotropic hormone (ACTH) treated rats. The administration of imipramine for 14 days decreased the (+/-) DOI-induced wet-dog shakes response; chronic administration of lithium for 14 days, however, had no effect. Chronic ACTH (100 microg/rat sc) treatment increased the wet-dog shake response induced by (+/-)-DOI. This effect of ACTH for 14 days, increasing the (+/-)-DOI-induced wet-dog shakes, was not inhibited by a 14-day administration of imipramine. Chronic coadministration of imipramine and lithium, lasting 14 days, decreased the wet-dog shakes response induced by (+/-)-DOI in rats treated with ACTH for 14 days. These findings indicate that lithium inhibits the hyperfunction of the 5-HT2A receptor in rats treated with ACTH when coadministered with imipramine. PMID- 11900812 TI - Role of 5-HT1A receptors in the control of food intake in obese Zucker rats of different ages. AB - The present study describes the role of 5-HT1A receptors in the serotonergic control of food intake in obese Zucker rats of different ages. In addition, serotonin (5-HT) and cholecystokinin (CCK) content and 5-HT turnover were determined in various brain regions. The 5-HT1A receptor agonist 8-hydroxy-2-(di n-propylamino)tetralin (8-OH-DPAT; 100 microg/kg) stimulated food intake in 3 month-old lean control rats but inhibited feeding in obese Zucker rats (300 microg/kg). This pattern remained the same in 6-month-old rats. At 10 months of age, 8-OH-DPAT lost its inhibitory activity in the obese rats but still stimulated feeding in lean controls (300 microg/kg). 5-HT levels were higher in the hypothalamus and in the frontal and parietal cortices of 3-month-old obese Zucker rats and were associated with a lower cortical turnover. In the parietal cortex and the hypothalamus of 6-month-old rats, 5-HT levels were still higher, linked with a lower hypothalamic turnover. No differences were observed in 10 month-old rats. CCK content was not different between obese Zucker rats and lean rats. The persistently different feeding responses to 8-OH-DPAT in obese Zucker rats and lean controls may be related to changes in brain 5-HT metabolism in the obese Zucker rats. PMID- 11900813 TI - Mouse lines differing in sensitivity to beta-CCM differ in tasks used for testing antidepressants. AB - Two lines of mice, previously selected for their sensitivity (BS) or their resistance (BR) to an anxiogenic benzodiazepine (BZ) receptor inverse agonist, methyl beta-carboline-3-carboxylate (beta-CCM), have recently been shown to present several differences in anxiety. In the present study, attempt was made to extend their behavioral profile in two situations classically used for testing antidepressant drugs. Reassessment of locomotor performance of these new populations confirmed that the motor activity of BR mice was lower than that of BS mice. In both the forced-swimming and the tail suspension tests, the immobility time of BS mice was significantly higher than that of BR mice. In the tail suspension test, two administrations of imipramine (30 mg/kg i.p., 5 h and 30 min before testing) significantly reduced the immobility time of BS mice but not of BR mice. From these data, it appears that BS mice are more "depressed" than BR mice. Thus, these selectively bred lines may represent potentially useful animal models to investigate behavioral, neurochemical and neuroendocrine correlates of antidepressant action. PMID- 11900814 TI - Tamoxifen and toremifene impair retrieval, but not acquisition, of spatial information processing in mice. AB - The present study examines the effects of tamoxifen (TAM) or toremifene (TOR), two triphenylethylene antiestrogen agents, on spatial information in mice by using Morris water maze. In a 30-s free swim trial, the TAM- or TOR-treated mice (intraperitoneally, 30 min before test) spent shorter time than the blank control mice in target quadrant. Compared to saline control group, animals exposed to TAM (1-10 mg/kg i.p., once a day for 5 days) or TOR (3-30 mg/kg i.p., once a day for 5 days) did not show significant difference on the acquisition of place task in Morris water maze. These results suggest that TAM, at the doses of 1-10 mg/kg, and TOR, at the doses of 3-30 mg/kg, impair the retrieval, but not the acquisition, of spatial information task in Morris water maze. It seems, however, that TOR is more potent than TAM on impairing memory retrieval. PMID- 11900815 TI - Acute third ventricular administration of insulin decreases food intake in two paradigms. AB - The pancreatic hormone, insulin, has been hypothesized to be an important regulator of food intake. Consistent with this hypothesis is the finding that exogenous insulin, in doses that do not affect blood glucose, reliably suppresses food intake and body weight. However, previous experiments have utilized a long term delivery paradigm, in which insulin is administered via osmotic minipump and changes in body weight and food intake are measured across days. In separate experiments, we report that acute central injections of insulin can reduce food intake. In Experiment 1, injection of insulin (8 mU) into the third cerebral ventricle reliably suppressed intake of pelleted rat chow beginning at onset of the rats' dark phase. In Experiment 2, central insulin reliably and dose dependently suppressed intake of a 1-h 15% sucrose meal in the middle of the light phase. These data suggest that insulin can reduce food intake in acute delivery paradigms and provide another means by which to assess the roles of other central systems in the mediation of insulin's effects on energy homeostasis. PMID- 11900816 TI - Effects of progesterone treatment on smoked cocaine response in women. AB - The effects of female sex hormones on responses to cocaine have not been systematically investigated in women. In this study, the safety and efficacy of acute progesterone treatment on smoked cocaine response was examined in female cocaine users. Five women had two experimental sessions during the early follicular phase, within 3-9 days after the beginning of their menses. In each experimental session, subjects received a single 200-mg dose of progesterone or placebo orally. Starting 2 h after the medication treatment, subjects received three deliveries of 0.4-mg/kg smoked cocaine 30 min apart. Progesterone treatment, compared to placebo, did not affect the blood pressure and heart rate changes in response to cocaine deliveries. For subjective responses to cocaine, the average of five-item Cocaine Effects Questionnaire (CEQ) was attenuated under progesterone treatment compared to placebo. For individual items of CEQ, progesterone treatment was associated with diminished rating of "feel the effect of last dose" in response to cocaine. These preliminary results suggest that acute progesterone treatment, given during the early follicular phase, may attenuate some of the subjective effects of cocaine. Further studies are warranted to examine the effects of progesterone treatment on cocaine dependence. PMID- 11900817 TI - Experimental anxiety induced by histaminergics in mast cell-deficient and congenitally normal mice. AB - To clarify the effect of mast cell-derived histamine release in the brain on anxiety, histaminergics-induced anxiety-like behaviors were examined by a light/dark test in mast cell-deficient (W/Wv) and congenitally normal (+/+) mice. In +/+ mice, when cimetidine (an H2 receptor antagonist) was coadministered with thioperamide (a neuronal histamine releaser acting via inhibition of H3 autoreceptors) or Compound 48/80 (C48/80, a selective histamine releaser from mast cells), the time spent in the light zone and the number of crossings between light and dark zones in a light/dark test decreased significantly, suggesting induction of anxiety. In W/Wv mice, however, experimental anxiety was induced by coadministration of thioperamide-cimetidine, but not C48/80-cimetidine. These results suggest that both nonneuronal mast cell-derived histamine and neuronal histamine play an important role in inducing experimental anxiety. PMID- 11900818 TI - Bidis--hand-rolled, Indian cigarettes: effects on physiological, biochemical and subjective measures. AB - Bidis, hand-rolled cigarettes imported from India, have become increasingly popular among US teenagers. These cigarettes are perceived as a safer, more natural alternative to conventional cigarette smoking. The present study was conducted to determine whether the acute effects of bidis and conventional cigarettes are similar. Undergraduate cigarette smokers with a history of bidi smoking were tested in two experimental sessions, using a within-subject design. Subjects smoked both a bidi and a conventional cigarette. Physiological and biochemical measures, subjective evaluations, and smoking behavior characteristics were obtained before, during, and after smoking each experimental cigarette. Although time to smoke and puffs per cigarette were significantly higher after the bidi, physiological and biochemical effects of bidi smoking were similar to those of smoking conventional cigarettes. Bidis were rated less satisfying than the conventional cigarette. However, there were no significant differences between the cigarettes in other subjective measures. Our results do not support the belief that bidis are a safe alternative to conventional cigarettes. Furthermore, bidi smoking, like conventional cigarette smoking, may lead to nicotine dependence. PMID- 11900819 TI - Prior maze experience required to alter midazolam effects in rats submitted to the elevated plus-maze. AB - In rodents, prior maze experience increases open arm avoidance (OAA) and compromises the anxiolytic effects of benzodiazepines in a subsequent exposure to the elevated plus-maze (EPM), a phenomenon referred to as "one trial tolerance" (OTT). Nevertheless, a possible correlation between these intriguing events remains unclear. Using maze-naive and maze-experienced (free exploration of the EPM for 5 min) rats, Experiment 1 confirmed the anxiolytic effects of midazolam (MDZ; 0.125-1.0 mg/kg) in maze-naive rats, while both increased OAA and OTT to the MDZ anxiolytic effects were observed in maze-experienced rats. However, our results from Experiment 2, designed to assess whether open, enclosed or both arms experience is involved in increased OAA and OTT, showed that MDZ retained its efficacy in rats confined either to an open or enclosed arm, where no significant changes in open arm exploration were observed when compared to the maze-naive group, therefore suggesting that prior experience in the whole apparatus may be involved in the loss of the anxiolytic MDZ effects. Results are discussed in terms of a possible correlation between increased OAA and the OTT phenomenon elicited in a subsequent exposure to the EPM. PMID- 11900820 TI - Acute nicotine effects on auditory sensory memory in tacrine-treated and nontreated patients with Alzheimer's disease: an event-related potential study. AB - The auditory mismatch negativity (MMN) event-related brain potential (ERP) reflects the storage of information in acoustic sensory memory. Thirteen patients with Alzheimer's disease (AD), 6 receiving treatment with the cholinesterase inhibitor, tacrine [tetrahydroaminoacridine (THA)], and 7 receiving no treatment, were administered 2 mg of nicotine polacrilex and placebo. MMNs were recorded with 1- and 3-s interstimulus intervals (ISIs) during pre- and post placebo/nicotine administration. Amplitudes decreased from pre- to post-placebo recordings in nontreated patients but remained stable in THA-treated patients. Comparison of pre- and post-nicotine MMNs found amplitude increases with nicotine in nontreated but not in THA-treated patients. MMN latencies were shortened by nicotine in both treatment groups. These exploratory findings suggest that nicotine-improved strength of acoustic sensory memory traces and speed of acoustic sensory discrimination in AD are differentially affected by chronic tacrine treatment. PMID- 11900821 TI - Behavioural and biochemical studies of citalopram and WAY 100635 in rat chronic mild stress model. AB - Reversal of chronic mild stress (CMS)-induced decrease of sucrose consumption has been studied in rats after 2, 7, 14, and 35 days treatment with imipramine, citalopram (both 10 mg/kg per day, i.p.), WAY 100635 (0.2 mg/kg sc, b.i.d.), and citalopram plus WAY 100635. Bmax, Kd, and functional status [cyclic AMP (cAMP) generation] of beta1-adrenoceptors were assessed in cortical tissue at the same time points. Citalopram reversed CMS-induced reduction of sucrose intake at an earlier time point than imipramine. WAY 100635 was not effective and did not potentiate the effect of citalopram. CMS produced increase of Bmax. Imipramine decreased Bmax in controls (Days 2, 7, 14, and 35) and normalised Bmax in stressed animals (Day 35). Citalopram, WAY 100635, and the combination increased Bmax in stressed animals and controls (Days 14 and 35). Inconsistent changes of Kd values and of cAMP responses to noradrenaline (NA) stimulation were observed. Thus stress- and drug-induced effects on beta1-adrenoceptors do not appear to be a common biochemical marker of antidepressant-like activity in the CMS model. PMID- 11900822 TI - Low alcohol preference among the "high alcohol preference" C57/BL10 mice; factors affecting such preference. AB - The effects of age, ethanol concentration and minor stress on the variation in alcohol preference of C57 strain mice were determined. In two bottle choice tests, an older population of mice contained slightly more low-preference mice than a younger population. A wide range of ethanol preference was consistently seen in young mice for 8% and 6% ethanol, but the previously reported biphasic pattern of distribution was revealed only with 8% ethanol. Very few animals showed high preference for concentrations of 10% or 12% ethanol. Moving low alcohol preference mice to a new location (but not repeated cage changing or ultrasonic noise) significantly increased the alcohol preference. Exploratory locomotor activity did not correlate with the subsequent alcohol consumption. Blood and brain alcohol concentrations showed that the differences in alcohol preference were not due to differences in metabolism of ethanol. The C57 strain mice with low preference for alcohol provides a valuable model for the study of the effects of minor stress on alcohol consumption. PMID- 11900823 TI - A comparison of the effects of 6-beta naltrexol and naltrexone on the consumption of ethanol or sucrose using a limited-access procedure in rats. AB - We recently reported that 6-beta naltrexol, the major metabolite of naltrexone in humans, reduced ethanol consumption in rats. Two new experiments were designed to compare 6-beta naltrexol and naltrexone across three dose levels on an ethanol or sucrose baseline using a limited-access procedure in Wistar rats. The results of Experiment 1 showed that both 6-beta naltrexol and naltrexone reduced ethanol consumption across a range of doses. An in vivo assay showed that naltrexone was approximately 25 times more potent than 6-beta naltrexol at comparable ED50 doses. In addition, there was no indication of systematic development of tolerance to the effect of either drug across the 4 days of drug administration. In Experiment 2, both 6-beta naltrexol and naltrexone reduced the consumption of a sucrose solution using a limited-access procedure. The implications of these data for the development of pharmacotherapeutic agents capable of reducing drinking in recovering alcoholics are discussed. PMID- 11900824 TI - Alcohol-preferring AA rats show a derangement in their central melanocortin signalling system. AB - The AA (Alko, Alcohol) rats are selectively bred for their preference of alcohol to water, contrasting to ANA rats that avoid alcohol. They also exhibit a lower growth rate compared to ANA rats, as well as differences in their response to substances affecting food intake. The melanocortin (MC) system is involved in the regulation of feeding behaviour and in mechanisms underlying drug addiction and tolerance. Recently, administration of an MC receptor agonist proved to reduce alcohol intake in AA rats. We predicted that the ratio of endogenous MC receptor agonists (proopiomelanocortin, POMC) and antagonists (agouti-related protein, AgRP) would differ from ANA rats, and that subsequent differences in MC receptor levels would be detectable. We used in situ hybridization to detect an increased ratio of POMC/AgRP mRNA in the arcuate nucleus (Arc) of AA rats. Receptor autoradiography indicated that MC3 receptor binding differed in the nucleus accumbens and several hypothalamic nuclei, possibly reflecting differences in MC peptide transmission in the AA rats. Our results support the claim that AA rats have a high ratio of POMC/AgRP expression, and that this observation is accompanied by differences in MC3 receptor levels. PMID- 11900825 TI - Visual cortical organization at the single axon level: a beginning. AB - Single axon analysis of visual cortical connections is an important extension of previous anterograde studies using 3H-amino acids or wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP). The higher resolution tracers Phaseolus vulgaris-leucoagglutinin (PHA-L), biocytin, biotinylated dextran amine (BDA) and dextran-conjugates-have already produced new results, simply by providing improved visualization, concerning laminar definition and possible subtypes of connections, as well as the beginning of a database of morphometrics and microstructure. The comparative approach, comparing geniculocortical terminations and cortical connections across several areas, has suggested both specific structural-functional correlations (for example, in extrastriate area MT/V5) and more subtle, possibly gradient-wise variations. Likely future directions for this line of research include more direct correlations of axon geometry with functional architectures, investigations of microcircuitry at the level of electron or confocal microscopy, anatomical and functional investigations of connectional convergence and interactions, and, not least, a more comprehensive database. PMID- 11900826 TI - Cellular and molecular basis for the formation of lamina-specific thalamocortical projections. AB - The neocortex is composed of a characteristic layered structure, which is a basis of extrinsic and intrinsic cortical connections. In recent years the cellular and molecular mechanisms, which are responsible for the formation of lamina-specific connections, have been explored by extensive molecular and in vitro studies. This article attempts to address what cell-cell interactions are required for axonal targeting and what molecules regulate cellular events, focusing upon the development of the thalamocortical projection. PMID- 11900827 TI - Effects of temperature increase on the propagation of presynaptic action potentials in the pathway between the Schaffer collaterals and hippocampal CA1 neurons. AB - Effects of temperature increase on the neuronal activity of hippocampal CA2-CA1 regions were examined by using optical and electrophysiological recording techniques. Stimulation of the Schaffer collaterals at the CA2 region evoked depolarizing optical signals that spread toward the CA1 region at 32 degrees C. The optical signal recorded by 49 pixels was characterized by fast and slow components that were closely related to presynaptic action potentials and excitatory postsynaptic responses, respectively. The optical signal was depressed by temperature increase to 38-40 degrees C. The temperature increase to 38 degrees C produced a hyperpolarization and a depression of the excitatory postsynaptic potential (EPSP) in single hippocampal CA1 pyramidal neurons. The depression of the neuronal activity induced by temperature increase was attenuated by application of glucose (22 mM) or pyruvate (22 mM). Adenosine (200 microM) did not block the presynaptic action potential but strongly depressed the excitatory postsynaptic response. 8-Cyclopentyl-1,3-dimethylxanthine (8-CPT) (10 microM), an antagonist for adenosine A(1) receptors, attenuated the depression of the excitatory postsynaptic response but not the inhibition of the presynaptic action potential at 38 degrees C. These results suggest that adenosine mediates the high-temperature-induced depression of the excitatory synaptic transmission but not that of action potential propagation in rat CA1 neurons. PMID- 11900828 TI - Focal stimulation of single GABAergic presynaptic boutons on the rat hippocampal neuron. AB - Evoked inhibitory postsynaptic currents (eIPSCs) generated from a single GABAergic bouton were recorded and the functional properties were investigated. Native single boutons attached to mechanically dissociated rat hippocampal CA1 neurons, namely "synaptic bouton" preparation, were visualized with FM 1-43 dye and selectively stimulated by a glass pipette directed to a single bouton by focal stimulation. The GABAergic eIPSCs were elicited in like all-or-none fashion regarding both stimulus strength and pipette location, thus indicating that the eIPSCs result from the activation of a single bouton. The GABA release from the boutons was action potential dependent since eIPSCs were blocked in the presence of either voltage-dependent Na(+) or Ca(2+)channel blocker. Even in the presence of tetrodotoxin (TTX), eIPSCs could be elicited by additional application of a voltage-dependent K(+) channel blocker, 4-AP. The GABA release depended on external Ca(2+) concentration. Amplitude histogram of eIPSCs did not follow Poisson distribution or show discrete peaks. As a result, this new experimental approach using both focal stimulation and a synaptic bouton preparation allows for a detailed study of the native synaptic machinery in nerve terminals measuring smaller than 1 microm in size in the CNS. PMID- 11900829 TI - Expression of galanin receptor-1 (GALR1) in the rat trigeminal ganglia and molar teeth. AB - The expression of galanin receptor-1 (GALR1) was investigated in the rat trigeminal ganglion by using immunocytochemistry and in situ hybridization. In addition, the regional distribution of GALR1-immunoreactive pulpal nerves and their ultrastructure were examined in the molar teeth. In the trigeminal ganglion, the immunoreactivity for GALR1 was recognizable in about 30% of the total number of neurons. Most of the cell bodies were small to medium in size. Analysis of serially cut sections alternately stained with GALR1 and galanin antisera demonstrated that some GALR1-positive cells displayed immunoreactivity for galanin. In situ hybridization analysis, expression of GALR1 mRNA was detected in trigeminal ganglion cells. The cell size distribution was similar to that of GALR1-immunoreactive cells. In the dental pulp, a small number of nerve fibers displayed immunoreactivity for GALR1. The labeled fibers formed terminal arbors in the coronal pulp around and within the odontoblast cell layer, but never penetrated into the predentin and dentin. Ultrastructurally, GALR1 immunoreactivity in the dental pulp was confined to the axoplasm of unmyelinated nerve fibers. The present study provided new evidence that unmyelinated primary afferents innervating dental pulp possessed galanin receptor, and suggests the existence of nociceptive primary afferents functioning as autocrine cells. PMID- 11900830 TI - Development of the rat respiratory neuron network during the late fetal period. AB - We studied developmental changes in respiratory-like C4 activity and respiratory related neurons in the ventrolateral medulla (VLM) of brainstem-spinal cord preparations from rat fetuses after embryonic day 16 (E16). In addition to respiratory nerve activity, non-respiratory activity was recorded from the C4 ventral root of preparations before E19. The burst duration of respiratory nerve discharge increased markedly at E19/20. Subtypes of neurons similar to newborn respiratory neurons were found in preparations with prolonged burst duration (more than 400 ms) after E20. These subtypes were not evident in preparations with short burst duration (less than 300 ms) before E19. About 60% of the inspiratory neurons in E17-19 preparations produced voltage-dependent burst activity, which was preserved in low Ca(2+)/high Mg(2+) synaptic blockade solution. In about 11% of the inspiratory neurons of E18-19 preparations, activation of one neuron induced activation of the inspiratory neuron network and generation of a full C4 inspiratory burst. The present findings suggest that respiratory neuron networks mature functionally to the level of the neonatal respiratory neuron networks during gestation period E19/20. Potentiation of synaptic interaction between respiratory neurons, causing developmental changes in the burst pattern, might be involved in the maturation process during late fetal stages. PMID- 11900831 TI - Connections between the amygdala and auditory cortical areas in the macaque monkey. AB - Connections between the amygdala and auditory cortical areas TC, and the rostral, intermediate and caudal regions of area TA (TAr, TAi and TAc, respectively) in the macaque monkey (Macaca fuscata and Macaca nemestrina) were investigated following placements of cortical deposits of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Areas TC and TAc received weak projections and these derived only from the lateral basal nucleus. Areas TAi and TAr received projections from the lateral, lateral basal and accessory basal nuclei. In contrast, corticopetal projections to the amygdala originated in areas TAi and TAr, but never in TAc or TC. The projections from areas TAi and TAr terminated only in the lateral nucleus, and in particular at the lateral part of the middle and caudal portions of the amygdala. Thus, the amygdalofugal projections to the auditory cortices are more widespread and more complex than the amygdalopetal projections of the auditory cortices. As judged from experiments in which deposits were made at different sites along the rostrocaudal axis of the auditory cortex, there was a progressive increase seen in density of the amygdala connections with more anteriorly-placed injection sites. PMID- 11900833 TI - Editorial. PMID- 11900832 TI - Bupivacaine, but not tetracaine, protects against the in vitro ischemic insult of rat hippocampal CA1 neurons. AB - Neuroprotective actions of local anesthetics, bupivacaine and tetracaine, against the irreversible membrane dysfunction induced by in vitro ischemia were investigated. Intracellular recordings were made from hippocampal CA1 neurons in rat brain slice preparations. Oxygen and glucose deprivation (in vitro ischemia) produced a rapid depolarization after approximately 5 min of exposure. When oxygen and glucose were reintroduced, the membrane depolarized further and reached at 0 mV: the membrane showed no functional recovery (irreversible membrane dysfunction). Pretreatment with tetracaine or bupivacaine significantly prolonged the latency of rapid depolarization. Bupivacaine, but not tetracaine, restored the membrane potential after the reintroduction of oxygen and glucose. Tetracaine and bupivacaine depressed both field postsynaptic potentials and presynaptic volleys. The drugs also reduced the dV/dt of Ca(2+)-dependent spikes and the rapid rise of [Ca(2+)](i) induced by in vitro ischemia. Compared with tetracaine, bupivacaine markedly suppressed the resting K(+) conductance and the ATP-sensitive and Ca(2+)-dependent K(+) conductances. Moreover, in the presence of tetraethylammonium (TEA), a majority of CA1 neurons impaled with Cs acetate filled electrodes showed complete or partial recovery of the membrane potential after reintroducing oxygen and glucose. These results suggest that the neuroprotective action of bupivacaine is mainly due to the suppression of the K(+) conductances. PMID- 11900834 TI - EHV-1 EICP22 protein sequences that mediate its physical interaction with the immediate-early protein are not sufficient to enhance the trans-activation activity of the IE protein. AB - The early 293 amino acid EICP22 protein (EICP22P) of equine herpesvirus 1 localizes within the nucleus and functions as an accessory regulatory protein (J. Virol. 68 (1994) 4329). Transient transfection assays indicated that although the EICP22P by itself only minimally trans-activates EHV-1 promoters, the EICP22P functions synergistically with the immediate-early protein (IEP) to enhance expression of EHV-1 early genes (J. Virol. 71 (1997) 1004). We previously showed that the EICP22 protein enhances the DNA-binding activity of the EHV-1 IEP and that it also physically interacts with the IEP (J. Virol. 74 (2000) 1425). In this communication, we employed transient trans-activation assays utilizing EICP22P deletion mutants to address whether the sequences required for EICP22P IEP physical interactions are essential for EICP22P's ability to interact synergistically with the IEP. Assays employing various classes of the EHV-1 promoters fused to the chloramphenicol acetyl-transferase (CAT) reporter gene indicated that: (1) neither full length nor any of the EICP22P mutants tested was able to overcome repression of the IE promoter elicited by the IEP, (2) the full length EICP22P interacted synergistically with the IEP to trans-activate the early and late promoters tested, and (3) all of the EICP22P mutants, including those that were able to physically interact with IEP and itself, failed to function synergistically with the IEP to trans-activate representative EHV-1 early and late promoters. The results suggest that EICP22P sequences required for its interaction with the IE protein are not sufficient to mediate its synergistic effect on the trans-activation function of the IEP. The possible explanations as to why sequences in addition to those that mediate EICP22P-IEP interaction and EICP22P self-interactions are essential for the synergistic function of EICP22P are discussed. PMID- 11900835 TI - Rapid evolution of two discrete regions of the caprine arthritis-encephalitis virus envelope surface glycoprotein during persistent infection. AB - Five major regions of sequence diversity between strains (V1-V5) have been described in the caprine arthritis-encephalitis lentivirus (CAEV) envelope surface unit glycoprotein (SU). To determine which of these variable regions is important in persistent infection in vivo, we evaluated SU sequence diversity in five neutralization variants from two goats and proviral DNA from five additional goats infected with CAEV-63 for up to 7 years. Overall amino acid sequence divergence in the SU encoded by provirus and neutralization variants compared to parental CAEV-63 ranged from 1.1 to 4%. However, most of the amino acid substitutions and all of the deletions and insertions were present in two discrete regions designated HV1 and HV2. The HV2 region was variable in all neutralization variants and provirus sequences from most animals. This region overlapped the V4 domain of CAEV SU and the neutralization domain of the closely related ovine maedi-visna lentivirus. HV1 was located in a region of SU strictly conserved in all small ruminant lentivirus strains except CAEV-63. This region only varied in a subset of neutralization variants and proviruses, all derived from goats with arthritis. In contrast, sequences in the V1,V2,V3, and V5 regions were stable in neutralization variants and proviruses from infected goats, indicating that sequence diversity between strains in these regions is not due to selection of variants in persistently infected animals. Our results define two discrete regions of CAEV SU that undergo rapid sequence variation in persistently infected goats which may have important roles in virus-host interactions. PMID- 11900836 TI - Virulent influenza A viruses induce apoptosis in chickens. AB - Virulent avian influenza A viruses produce lethal disease in chickens. Since cell death can be caused by either necrosis or apoptosis, we investigated the types of cell death that occur in natural hosts, chickens, infected with virulent avian viruses. Using biochemical methods, we demonstrate that virulent avian influenza viruses induce apoptosis of vascular endothelial cells in liver, kidney, and brain. Viral antigens were also detected in these organs, suggesting that viral replication induces apoptosis in infected chickens. These results indicate that apoptosis does occur in virulent avian influenza virus infection in a natural host, and may contribute to the lethality of the virus. PMID- 11900837 TI - Nucleo-cytoplasmic localization of influenza virus nucleoprotein depends on cell density and phosphorylation. AB - Influenza virus nucleoprotein (NP) plays a major role in the nucleus during virus replication, and is a mediator of viral ribonucleoprotein nuclear import during entry. NP is localized primarily in the nucleus, but can undergo nucleo cytoplasmic shuttling in heterokaryons (Whittaker et al., 1996a. J. Virol. 70, p. 2743). We have studied NP localization using a stable cell line (3PNP-4) that expresses NP. Intracellular localization of NP was markedly affected by the density of the cell monolayer. It was nuclear in cells grown in sparse culture, but cytoplasmic in dense culture. In phorbol ester-stimulated cells NP was cytoplasmic, but relocalized to the nucleus after treatment with a protein kinase inhibitor. Cell density and phosphorylation-dependent localization of NP appeared to be independent of cell type. Our data suggest that a phosphorylation event is needed either for nuclear export, or to regulate retention of NP in the nucleus, and that regulation may be mediated by kinases activated by cell-cell contact. PMID- 11900838 TI - Characteristics of a respiratory syncytial virus persistently infected macrophage like culture. AB - A persistently infected culture obtained from immortalized murine macrophage-like cells, which survived respiratory syncytial virus (RSV) infection at multiplicity of one, was established and characterized. The presence of RSV through the passages was confirmed and monitored by (a) detection of infectious virus by TCID(50)/ml, (b) defective particles by viral infectivity interference and buoyant density determinations, (c) cell surface antigen by indirect immunofluorescence and FACS, and (d) expression of a viral gene by RT-PCR. Moreover, cell morphology changes by comparison of macrophage area and perimeter were determined. A second culture was obtained by cell cloning out of this culture, and a third culture was established by superinfection with the original virus, in which 92-95% of the macrophages expressed viral antigen without cell destruction and released defective particles but low levels of infectious virus. Although the three cultures maintained the characteristics of persistently infected cells, concentrations of released infectious virus, defective particles, and percentages of cells bearing viral antigen varied. RSV persistently infected murine macrophage cultures provide an in vitro model to study viral-macrophage interaction and to allow the experimental use of a cell important in disseminating the infection. In addition, due to the wide array of cellular and humoral reagents in the mouse, studies on immunologic aspects of viral immunity are facilitated. PMID- 11900839 TI - Variation in the NS3/NS3A gene of bluetongue viruses contained in Culicoides sonorensis collected from a single site in southern California. AB - To determine the variability of the NS3/NS3A gene of field strains of BTV contained in Culicoides sonorensis collected from a single site in California (CA), the NS3/NS3A gene was directly amplified and sequenced from 22 pools of C. sonorensis and compared with those of previously characterized field isolates from CA, as well as to viruses that caused recent outbreaks of bluetongue disease in ruminants in CA. Phylogenetic analysis established that the NS3/NS3A gene of strains of BTV contained in C. sonorensis collected from the site exists as a heterogeneous population. The two most divergent nucleotide sequences of the NS3/NS3A genes of these viruses differed by 2.5% (18 nucleotides). Comparison with the NS3/NS3A gene sequences from viruses that caused recent instances of bluetongue disease in ruminants in CA indicated that BTV strains from different geographic regions can exhibit a higher degree of genetic heterogeneity (up to 6.6%; 0-48 nucleotide differences) than those contained in C. sonorensis collected from a single site. PMID- 11900840 TI - Generation of recombinant vaccinia viruses expressing Puumala virus proteins and use in isolating cytotoxic T cells specific for Puumala virus. AB - Puumala (PUU) virus causes a form of hemorrhagic fever with renal syndrome (HFRS), called nephropathia epidemica (NE), in Europe. HFRS is characterized by an increased capillary permeability, which we hypothesize is caused by hyperactivation of the host immune system, especially cellular immune responses. To identify cytotoxic T lymphocytes (CTLs) specific for the PUU virus from NE patients, we have made recombinant vaccinia viruses expressing PUU virus proteins, the nucleocapsid (N) and two surface glycoproteins, G1 and G2. Recombinant vaccinia viruses carrying the N or the first half of the G2 cDNA under the control of a strong synthetic promoter were made. To express G1 and the second half of the G2 proteins, however, we needed to use a T7 expression system, where the T7 RNA polymerase is produced from another recombinant vaccinia virus co-infecting the same cells. These recombinant vaccinia viruses were used to detect and clone PUU virus-specific CTLs from the peripheral blood mononuclear cells of NE patients. An HLA-A24-restricted CTL line recognizing the G2 protein was isolated and its 9-mer epitope was determined. PMID- 11900841 TI - Pseudorabies virus (PRV) is protected from complement attack by cellular factors and glycoprotein C (gC). AB - Swine kidney derived CPK cells were resistant to swine complement attack in vitro while rabbit kidney derived RK13 cells were destroyed by swine complement. To rabbit complement, RK13 cells were resistant but CPK cells were sensitive. This phenomenon was known as homologous restriction (Proc. Natl. Acad. Sci. USA 78 (1981) 5118). The gC deletion mutant of pseudorabies virus (PRVdlgC) grown in CPK cells was resistant to swine complement while the same virus grown in RK13 cells was neutralized by swine complement. PRVdlgC grown in RK13 cells was more resistant to rabbit complement than the virus grown in CPK cells. Hence, the sensitivity of PRVdlgC to swine or rabbit complement was similar to that of the cells in which the virus was grown. It would appear that cell derived factors were present on the virion and they were protective against homologous complement but not against heterologous complement. The expression of gC rendered PRV more resistant to swine or rabbit complement, but the protective effect of gC was much less than that of cell derived factors. The best protection against complement was obtained when gC and cell derived factors were coexistent on the virion. PMID- 11900842 TI - Determination of the complete genomic sequence and analysis of the gene products of the virus of Spring Viremia of Carp, a fish rhabdovirus. AB - The complete genome of spring viremia of carp virus (SVCV) was cloned and the sequence of 11019 nucleotides was determined. It contains five open reading frames (ORF's) encoding for the nucleoprotein N; phosphoprotein P; matrix protein M; glycoprotein G; and the viral RNA dependent RNA polymerase L. Genes are organised in the order typical for rhabdoviruses: 3'-N-P-M-G-L-5'. The short leader and trailer regions of SVCV exhibit inverse complementarity and are similar to the respective 3' and 5' ends of the genome of vesicular stomatitis virus. To verify the predicted open reading frames proteins were expressed in bacteria and analysed with a polyclonal anti-SVCV serum. Furthermore, monospecific antisera against the distinct viral proteins were generated. Comparison of genome and protein confirm the assignment of SVCV to the genus Vesiculovirus. PMID- 11900843 TI - Molecular analysis of the S1 subunit of the spike glycoprotein of respiratory and enteric bovine coronavirus isolates. AB - It is unclear whether respiratory and enteric bovine coronavirus (BoCV) strains are distinctive in biological, antigenic and genetic characteristics. In the present study, we analyzed the nucleotide and amino acid sequence of the S1 subunit of the S glycoprotein, including the cleavage site, of both respiratory (n=5) and enteric (n=3) BoCV isolates including two paired isolates from the same feedlot animals and compared them with the prototype Mebus and two enteric and one respiratory BoCV strains from Quebec. A total of 75 polymorphic nucleotides were identified in the S1 subunit of the spike glycoprotein of BoCV isolates compared with the Mebus strain. These polymorphisms led to 42 amino acid changes at 38 distinct sites. The amino acid changes were distributed throughout the S1 subunit with clustering around residues 40-118, 146-179, and 458-531. Among these variations, only 19 amino acid substitutions altered the charge, hydrophobicity and surface probability of the protein. Based on phylogenetic analysis, our respiratory and enteric isolates clustered into two major groups with two subgroups. Although, there were only a few amino acid changes between the respiratory and enteric paired isolates, the other two respiratory isolates, one isolated from the same farm as a paired strain and the other from a different farm, showed more sequence diversity. Amino acid alterations in residues 113, 115, 118, 146, 148, 501, 510 and 531 of respiratory isolates conferred significant changes in the predicted secondary structure compared with the prototype winter dysentery (WD) and the calf diarrhea (CD) strains of BoCV. In conclusion, the data suggests that respiratory strains of BoCV may differ genetically from the classical calf enteric and adult WD strains. PMID- 11900844 TI - A single amino acid is critical for the expression of B-cell epitopes on the helicase domain of the pestivirus NS3 protein. AB - Truncated NS3 proteins, expressed by recombinant baculoviruses, were used to investigate the location of conserved B-cell epitopes on this non-structural bovine viral diarrhoea virus (BVDV) protein. A goat anti-pestivirus antiserum, and a panel of anti-NS3 monoclonal antibodies, including the BVDV-1 specific antibody P1D8, were used to verify the presence or absence of the epitopes. Interestingly, the monoclonal antibodies reacted only with the truncated protein encompassing the helicase domain of NS3. Expression of the B-cell epitopes was dependent on, but not within, a 57 amino acid sequence at the carboxy-terminal end of this protein, supporting observations that these conserved epitopes are conformational in nature. A comparison of deduced amino acid sequences of the helicase domain from BVDV-1, BVDV-2, BDV and CSFV isolates highlighted a single amino acid that appeared to be unique to P1D8-reactive BVDV-1 isolates. Site directed mutagenesis studies confirmed that this amino acid is critical for the expression of the BVDV-1 specific NS3 epitope recognised by the P1D8 monoclonal antibody. Surprisingly, the amino acid was also important for an epitope recognised by two group-specific monoclonal antibodies, P1H11 and P4A11. Protein modelling studies, based on the structure of the hepatitis C NS3 helicase domain, indicated that this amino acid occupies a prominent position on the surface of the protein. PMID- 11900845 TI - Transcriptional and translational expression kinetics of the bovine herpesvirus 1 UL51 homologue gene. AB - We characterized the expression kinetics of the transcript and protein generated from the bovine herpesvirus 1 (BHV1) homologue of the herpes simplex virus 1 (HSV1) UL51 gene. The BHV1 UL51 ORF, located at positions 7236-->7967 of the viral genome, generated a major 1.05 kb transcript accumulating at very low abundance as soon as 3 h post-infection (p.i.), after which its levels increased to reach a plateau from 6 to 12 h p.i., and then slowly decreased up to 24 h p.i. As determined by S1 nuclease protection assays, UL51 transcription initiated at two distinct sites located at 191 and 196 bases upstream from the initiation codon, corresponding to positions 7045 and 7040 of the viral genome, respectively. Western blotting of BHV1-infected protein cell lysates, using a BHV1-specific antiserum generated against a recombinant protein expressed in Escherichia coli, detected a 28 kDa protein of the expected size (24985 Da) whose expression kinetics followed that of its transcript. As evidenced by in situ immunofluorescence assays, the protein mainly localized to the cytoplasm and the perinuclear region of infected cells. In contrast to HSV1 UL51 which is classified as a gamma2 gene, BHV1 UL51 belongs to viral genes of the gamma1 class as expression of its transcript is partially dependent on viral DNA synthesis. PMID- 11900846 TI - Genetic characterization and phylogeny of human T-cell lymphotropic virus type I from Chile. AB - Infection with Human T-Cell Lymphotropic Virus type I (HTLV-I) have been associated with the development of the HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP). Phylogenetic analyses of HTLV-I isolates have revealed that HTLV-I can be classified into three major groups: the Cosmopolitan, Central African and Melanesian. In the present study, we analyzed the tax, 5' ltr, gag, pol, and env sequences of proviruses of PBMC from ten HAM/TSP patients to investigate the phylogenetic characterization of HTLV-I in Chilean patients. HTLV-I provirus in PBMC from ten Chilean patients with HAM/TSP were amplified by PCR using primers of tax, 5' ltr, gag, pol, and env genes. Amplified products of the five genes were purified and nucleotide sequence was determined by the dideoxy termination procedure. DNA sequences were aligned with the CLUSTAL W program. The results of this study showed that the tax, 5' ltr, gag, pol, and env gene of the Chilean HTLV-I strains had a nucleotide homology ranged from 98.1 to 100%, 95 to 97%, 98.9 to 100%, 94 to 98%, and 94.2 to 98.5% respect to ATK-1 clone, respectively. According to molecular phylogeny with 5' ltr gene, the Chilean HTLV-I strains were grouped with each other suggesting one cluster included in Transcontinental subgroup. PMID- 11900847 TI - Detection of oyster herpesvirus DNA and proteins in asymptomatic Crassostrea gigas adults. AB - Since 1972, several herpes-like virus infections have been reported among different bivalve species around the world. Most of these reports involved larvae or juveniles presenting high mortalities. Two case reports of herpes-like viruses concerned adult oysters, Crassostrea virginica in USA and Ostrea angasi in Australia. Molecular techniques including PCR and in situ hybridization (ISH) have been recently developed to detect the oyster herpesvirus genome. In the present study, 30 Pacific oyster, Crassostrea gigas, adults have been analyzed using three different techniques: PCR, ISH and immunochemistry, in order to detect herpesviruses in asymptomatic individuals. PCR and ISH allowed detection of oyster herpesvirus DNA in 93.3 and 86.6%, respectively, of analyzed oysters while polyclonal antibodies allowed detection of viral proteins in 76.6% of analyzed adult oysters. These results suggest that oyster herpesvirus infects adult oysters with high prevalence and that the virus may persist in its host after primary infection. The detection of viral DNA and viral proteins in the gonad of several individuals supports the hypothesis of a possible vertical transmission of the infection. Lastly, concordance among the three techniques used in this study is discussed. PMID- 11900848 TI - Identification and characterisation of the genomic segment 7 of the infectious salmon anaemia virus genome. AB - The isolation and characterisation of a gene encoding the putative matrix proteins of infectious salmon anaemia virus (ISAV) is reported. Following identification of an ISAV-specific sequence from a cDNA library, RACE-PCR was used to identify a mRNA transcript of approximately 1.2 kb containing the ISAV consensus sequence GCTAAGA at the 5' end. Although the cDNA transcript and its putative protein product did not possess high homology with other orthomyxoviral sequences, similarity to a paramyxovirus fusion glycoprotein and viral cell surface proteins was identified. The size of this transcript suggested that it was derived from segment 7 of the ISAV genome and encoded the matrix proteins. Like syntenic segments of other orthomyxoviruses, this segment was shown to encode at least two matrix proteins, M1 and M2. The existence of M1 and M2 ISAV mRNA was demonstrated by RT-PCR and sequencing, with the M1 transcript being more abundant than the M2 in infected cell cultures, as is found in other orthomyxoviruses. Nucleotide sequence comparison of segment 7 of the ISAV genome from isolates of different geographic origin indicated it to be the one of the most variable of the ISAV genes characterised to date. PMID- 11900849 TI - Transcriptional activation of the promoter of human cytomegalovirus immediate early gene (CMV-IE) by the hepatitis B viral X protein (HBx) through the NF kappaB site. AB - The reactivation of latent cytomegalovirus (CMV) in a human by another viral infection may induce virus-related symptoms. Based on this presumption, we investigated the effect of HBx on the activation of the CMV-IE, which is also known as a transactivator and potential oncogene. The HBx transactivated the CMV IE promoter by up to 4- and 18-fold factors in human liver HepG2 and lung fibroblast MRC-5 cells, respectively. Cotransfection of HBx with each transcription factor presented in the CMV-IE promoter showed that only NF-kappaB synergistically activated the promoter by up to a 14-fold factor. Serial deletion assays and point mutation analysis showed that the third NF-kappaB site (nt -267 to -258) and the second one (nt -162 to -153) appeared as the major responsible site and minor one, respectively, for the transactivation. These results suggest the possibility that the HBV infection of a cell previously infected by CMV would exert influence on the reactivation of the latent cytomegalovirus in a human to induce virus-related symptoms. PMID- 11900850 TI - Characterization of the murine cytomegalovirus 38 kDa m137 gene product. AB - Murine cytomegalovirus (MCMV) m137 null mutants, Deltam137A and Deltam137B, were generated by inserting a gpt cassette into a deleted region of the open reading frame. A polyclonal antiserum produced to an Escherichia coli expressed gst-m137 fusion protein was used to show that a 38 kDa polypeptide corresponding to the predicted m137 gene product was present in NIH 3T3 fibroblasts infected with wild type MCMV but was not detected in Deltam137 infected cells. The protein did not fractionate with infected cell membranes and was not detectable in purified wild type virions. Plaque size, plaque morphology, and viral yield did not differ significantly between Deltam137 and wild-type MCMV infected 3T3 fibroblasts. The results showed that deletion of the 38 kDa protein did not negatively effect viral growth in 3T3 fibroblasts indicating that the m137 gene product is not essential for replication in these cells. In vivo analysis revealed that two independently isolated m137 mutants showed a significant delay in time until death but ultimately killed 100% of the mice in a SCID mouse model of virulence. PMID- 11900853 TI - Increased sensitivity to NMDA is involved in alcohol-withdrawal induced cytotoxicity observed in primary cultures of cortical neurones chronically pre treated with ethanol. AB - Severe cellular damage and neuronal cell loss were previously observed in cultures of primary cortical neurones after chronic ethanol pre-treatment followed by ethanol-withdrawal. In this study, we investigated the circumstances and the possible cellular changes leading to alcohol-withdrawal induced neuronal cell death. When cultures were pre-treated with ethanol (25-200mM) once for 24 or 72h, the amount of the subsequent 24h alcohol-withdrawal induced cell death estimated by measuring the release of lactate dehydrogenase (LDH)-was elevated only in cultures pre-treated with 200mM ethanol for 72h. On the contrary, as little as 50mM ethanol produced significant (P<0.01) increase in the withdrawal induced LDH-release in cultures pre-treated repeatedly with ethanol once daily for three consecutive days. When ethanol was re-added to the cultures during the withdrawal period, the LDH-release was dose-dependently reduced to the level of control. In ethanol pre-treated cultures N-methyl-D-aspartate (NMDA) (0.01-1mM) induced excitotoxicity as well as NMDA evoked elevation of cytosolic calcium ion concentration was increased. In contrast, the depolarising agent veratridine (0.01-1mM) produced similar extent of neuronal injury and elevation in cytosolic calcium ion concentration in control as in ethanol pre-treated cultures. According to these observations, repeated ethanol treatment appears to cause more robust adaptive changes in cultured neurones leading to more pronounced withdrawal induced cellular damage than chronic but single treatment does. In addition, the glutamatergic neurotransmission, especially the NMDA receptor system seems to be highly involved in the adaptive changes and in the cytotoxic effect of alcohol-withdrawal. PMID- 11900854 TI - Inhibition of the mitochondrial respiratory chain complex activities in rat cerebral cortex by methylmalonic acid. AB - Propionic and methylmalonic acidemic patients have severe neurologic symptoms whose etiopathogeny is still obscure. Since increase of lactic acid is detected in the urine of these patients, especially during metabolic decompensation when high concentrations of methylmalonate (MMA) and propionate (PA) are produced, it is possible that cellular respiration may be impaired in these individuals. Therefore, we investigated the effects of MMA and PA (1, 2.5 and 5mM), the principal metabolites which accumulate in these conditions, on the mitochondrial respiratory chain complex activities succinate: 2,6-dichloroindophenol (DCIP) oxireductase (complex II); succinate: cytochrome c oxireductase (complexII+CoQ+III); NADH: cytochrome c oxireductase (complex I+CoQ+complex III); and cytochrome c oxidase (COX) (complex IV) from cerebral cortex homogenates of young rats. The effect of MMA on ubiquinol: cytochrome c oxireductase (complex III) and NADH: ubiquinone oxireductase (complex I) activities was also tested. Control groups did not contain MMA and PA in the incubation medium. MMA significantly inhibited complex I+III (32-46%), complex I (61-72%), and complex II+III (15-26%), without affecting significantly the activities of complexes II, III and IV. However, by using 1mM succinate in the assay instead of the usual 16mM concentration, MMA was able to significantly inhibit complex II activity in the brain homogenates. In contrast, PA did not affect any of these mitochondrial enzyme activities. The effect of MMA and PA on succinate: phenazine oxireductase (soluble succinate dehydrogenase (SDH)) was also measured in mitochondrial preparations. The results showed significant inhibition of the soluble SDH activity by MMA (11-27%) in purified mitochondrial fractions. Thus, if the in vitro inhibition of the oxidative phosphorylation system is also expressed under in vivo conditions, a deficit of brain energy production might explain some of the neurological abnormalities found in patients with methylmalonic acidemia (MMAemia) and be responsible for the lactic acidemia/aciduria identified in some of them. PMID- 11900855 TI - The inhibition of tryptophan hydroxylase, not protein synthesis, reduces the brain trapping of alpha-methyl-L-tryptophan: an autoradiographic study. AB - The effects of the tryptophan hydroxylase (TPH) inhibitor p-chlorophenylalanine (PCPA; 200mg/kg; 3 days), and of the protein synthesis inhibitor cycloheximide (CXM, 2mg/kg), on regional serotonin (5-HT) synthesis were studied using the alpha-[14C]methyl-L-tryptophan (alpha-[14C]MTrp) autoradiographic method. The objectives of these investigations were to evaluate the changes, if any, on 5-HT synthesis, as measured with alpha-MTrp method, following the inhibition of TPH by PCPA, or the inhibition of proteins synthesis by CXM. The rats were used in the tracer experiment approximately 24h after the last dose of PCPA was administered, and in the CXM experiments, they were used 30 min following a single injection of CXM. In both experiments, the control rats were injected with the same volume of saline (0.5 ml/kg; s.c.) and at the same times as the drug injections. The results demonstrate that trapping of alpha-MTrp, which is taken to be related to brain 5-HT synthesis, is drastically reduced (40-80%) following PCPA treatment. The inhibition of protein synthesis with CXM did not have a significant effect on the global brain trapping of alpha-MTrp and 5-HT synthesis. These findings suggest that the brain trapping of alpha-[14C]MTrp relates to brain 5-HT synthesis, but not to brain protein synthesis. PMID- 11900856 TI - Phenylethanolamine N-methyltransferase has beta-carboline 2N-methyltransferase activity: hypothetical relevance to Parkinson's disease. AB - Mammalian brain has a beta-carboline 2N-methyltransferase activity that converts beta-carbolines, such as norharman and harman, into 2N-methylated beta carbolinium cations, which are structural and functional analogs of the Parkinsonian-inducing toxin 1-methyl-4-phenylpyridinium cation (MPP+). The identity and physiological function of this beta-carboline 2N-methylation activity was previously unknown. We report pharmacological and biochemical evidence that phenylethanolamine N-methyltransferase (EC 2.1.1.28) has beta carboline 2N-methyltransferase activity. Specifically, purified phenylethanolamine N-methyltransferase (PNMT) catalyzes the 2N-methylation (21.1 pmol/h per unit PNMT) of 9-methylnorharman, but not the 9N-methylation of 2 methylnorharmanium cation. LY134046, a selective inhibitor of phenylethanolamine N-methyltransferase, inhibits (IC50 1.9 microM) the 2N-methylation of 9 methylnorharman, a substrate for beta-carboline 2N-methyltransferase. Substrates of phenylethanolamine N-methyltransferase also inhibit beta-carboline 2N methyltransferase activity in a concentration-dependent manner. beta-Carboline 2N methyltransferase activity (43.7pmol/h/mg protein) is present in human adrenal medulla, a tissue with high phenylethanolamine N-methyltransferase activity. We are investigating the potential role of N-methylated beta-carbolinium cations in the pathogenesis of idiopathic Parkinson's disease. Presuming that phenylethanolamine N-methyltransferase activity forms toxic 2N-methylated beta carbolinium cations, we propose a novel hypothesis regarding Parkinson's disease a hypothesis that includes a role for phenylethanolamine N-methyltransferase catalyzed formation of MPP+ -like 2N-methylated beta-carbolinium cations. PMID- 11900857 TI - Quinolinic acid stimulates synaptosomal glutamate release and inhibits glutamate uptake into astrocytes. AB - Quinolinic acid (QA) is an endogenous neurotoxin involved in various neurological diseases, whose action seems to be exerted via glutamatergic receptors. However, the exact mechanism responsible for the neurotoxicity of QA is far from being understood. We have previously reported that QA inhibits vesicular glutamate uptake. In this work, investigating the effects of QA on the glutamatergic system from rat brain, we have demonstrated that QA (from 0.1 to 10mM) had no effect on synaptosomal L-[3H]glutamate uptake. The effect of QA on glutamate release in basal (physiological K+ concentration) or depolarized (40 mM KCl) conditions was evaluated. QA did not alter K+-stimulated glutamate release, but 5 and 10mM QA significantly increased basal glutamate release. The effect of dizolcipine (MK 801), a noncompetitive antagonist of N-methyl-D-aspartate (NMDA) receptor on glutamate release was investigated. MK-801 (5 microM) did not alter glutamate release per se, but completely abolished the QA-induced glutamate release. NMDA (50 microM) also stimulated glutamate release, without altering QA-induced glutamate release, suggesting that QA effects were exerted via NMDA receptors. QA (5 and 10mM) decreased glutamate uptake into astrocyte cell cultures. Enhanced synaptosomal glutamate release, associated with inhibition of glutamate uptake into astrocytes induced by QA could contribute to increase extracellular glutamate concentrations which ultimately lead to overstimulation of the glutamatergic system. These data provide additional evidence that neurotoxicity of QA may be also related to disturbances on the glutamatergic transport system, which could result in the neurological manifestations observed when this organic acid accumulates in the brain. PMID- 11900858 TI - Evidence for swelling-induced adenosine and adenine nucleotide release in rat cerebral cortex exposed to monocarboxylate-containing or hypotonic artificial cerebrospinal fluids. AB - Recent reports have described a swelling-induced release of adenosine triphosphate (ATP) from a variety of non-nervous system cell types, which may be involved in the regulatory volume decrease (RVD) response. The present study examined the effects of swelling induced by applications of hypotonic or monocarboxylic acid containing artificial cerebrospinal fluid (aCSF) on the release of adenosine nucleotides and adenosine from the in vivo rat cerebral cortex using a cortical cup technique. Hypotonic aCSF (25mM NaCl) elicited a significant increase in adenosine, but not adenine nucleotide, release. Applications of sodium L-lactate, pyruvate, or acetate (all 20mM) evoked increases in adenine nucleotides but not adenosine. D-Lactate (20mM) enhanced adenosine and ATP release. Inhibition of the plasma membrane monocarboxylate transporter with cyano-4-hydroxycinnamate (4-CIN, 2mM) blocked the effects of L lactate on purine release. These in vivo results demonstrate that osmoregulatory processes in cortical cells evoke an efflux of adenine nucleotides and/or adenosine. In that these purines activate a variety of receptors, it is possible that they may function as autocrine or paracrine signaling agents, facilitating volume regulation and enhancing local blood flow. PMID- 11900859 TI - The diversity of p53 mutations among human brain tumors and their functional consequences. AB - The p53 tumor suppressor is implicated in cell cycle control, DNA repair, replicative senescence and programmed cell death. Inactivation of the p53 contributes to the wide range of human tumors, including glial neoplasms. In this review, we describe the regulation and biochemical properties of p53 protein that may explain its ability to activate various genetic programs underlying cellular responses to stress conditions. The overall spectrum of p53 mutations is rather shared between tumor types indicating that these mutations are not tumor type specific. However, there is one example of germ-line mutation of p53 gene (the deletion of the codon 236) that is associated with a familiar brain tumor syndrome. We compare the frequency and type of most common mutations among various brain tumours (focusing on glioblastomas) and their consequences on protein functions. Furthermore, we discuss the most promising approaches of potential brain tumor therapy, including an adenovirus-mediated p53 gene transfer. Human glioblastomas are highly sensitive to the effects of p53 activity when the wild-type p53 is introduced ectopically. It suggests that the genetic or pharmacological modulation of the p53 pathway is potentially important strategy in the treatment of human cancers. PMID- 11900860 TI - BN52021, a platelet activating factor antagonist, is a selective blocker of glycine-gated chloride channel. AB - We have found that the platelet activating factor antagonist (BN52021) is an effective blocker of the glycine (Gly) receptor-mediated responses in the hippocampal pyramidal neurons of rat. Using the whole-cell voltage clamp and concentration clamp recording techniques, we investigated the mechanism underlying the inhibitory action of this terpenoid on the glycine-induced chloride current. BN52021 selectively and reversibly inhibits glycine current in a non-competitive and voltage-dependent fashion. The antagonistic effect of this substance is more pronounced at positive membrane potentials. At holding potential -70mV and in the presence of 200 microM glycine IC50 value for the blocking action of BN52021 was 270+/-10nM. Repetitive applications of BN52021 reveal the use-dependence of its blocking action. When co-applied with strychnine (STR), a competitive glycine receptor antagonist, BN52021 does not alter the IC50 value for strychnine. The inhibitory effect of BN52021 on gamma-aminobutyric acid (GABA) current is at least 25 times less potent than the effect on glycine current. This substance fails to affect AMPA and NMDA responses. It may be concluded that BN52021 inhibits glycine-gated Cl- channels by interacting with the pore region and does not compete for the strychnine-binding centre. PMID- 11900862 TI - Developmental expression and activity of high affinity glutamate transporters in rat cortical primary cultures. AB - The expression and activity of glutamate transporters (EAAC1, GLAST and GLT1) were examined during the development of cortical neuron-enriched cultures. Protein content and mitochondrial respiration both increased during the first 7 days, later stabilized and decreased from DIV14. Glutamate transport and extracellular concentration were relatively constant from DIV3 to 18. The kinetic parameters of glutamate transport were at DIV7: K(m)=19+/-3 microM and V(max)=1068+/-83 pmol/mg protein/min and at DIV14: K(m)=40.8+/-9.3 microM and V(max)=1060+/-235 pmol/mg protein/min. The shift in K(m) towards higher values suggest a more important participation of GLAST after DIV14. At DIV7 and 14, glutamate transport was poorly sensitive to dihydrokainate (DHK) suggesting a weak participation of GLT1 in glutamate transport. Western blot experiments and immunocytochemistry showed that EAAC1 was expressed by neurons whatever the stage of the culture. GLAST was found in astrocytes as soon as DIV3 and labeling increased during the development of the culture. There was little neuronal GLT1 immunoreactivity at DIV7, only detected by immunocytochemistry. From DIV10 to 18, an increasing astrocytic expression of GLT1 was observed, also detected by Western blotting. These results show that: (1) glutamate uptake remains stable all along the development of the cultures although the pattern of expression of the different transporters is changing, suggesting that glutamate transport is highly regulated; (2) neuronal EAAC1 may play a critical role during the early stages of the culture when it is expressed alone; and (3) the developmental expression pattern of glutamate transporters in cortical neuron-enriched cultures is quite similar to that observed in vivo during early postnatal development. PMID- 11900861 TI - Presynaptic inhibitory effects of rocuronium and SZ1677 on [3H]acetylcholine release from the mouse hemidiaphragm preparation. AB - It has been shown that nondepolarizing muscle relaxants may have effects on nicotinic acetylcholine receptors (nAChRs) other than those located on the skeletal muscle: some of them possess inhibitory effects on neuronal nAChRs [Anesth. Analg. 59 (1980) 935; Trends Pharmacol. Sci. 9 (1988) 16; Pharmacol. Ther. 73 (1997) 75]. It was shown that, e.g. (+)-tubocurarine and pancuronium are able to inhibit ACh release from the axon terminals of hemidiaphragm preparations and produce tetanic fade indicating their presynaptic effect. In this study rocuronium, a nondepolarizing steroidal muscle relaxant with shorter onset of action, and SZ1677 [1-(3alpha-hydroxy-17beta-acetyloxy)-2beta-(1.4-dioxa-8 azaspiro-[4,5]-dec-8-yl)-(5alpha-androstane-16beta-yl)-1-(2-propenyl) pyrrolidinium bromide], a short-acting muscle relaxant [Ann. New York Acad. Sci. 757 (1995b) 84] inhibited the release of ACh in response to axonal stimulation, while alpha-bungarotoxin failed to reduce the stimulation evoked release of ACh and did not produce tetanic fade. These results indicate that in addition to their postsynaptic effect, rocuronium and SZ1677 have presynaptic inhibitory effects on neuronal nAChRs at the neuromuscular junction. The finding that alpha bungarotoxin does not inhibit the release and does not produce tetanic fade indicates that it possesses affinity only for the postsynaptic muscle nAChRs. PMID- 11900864 TI - Synthesis, characterisation and biological activity of novel 4-thiazolidinones, 1,3,4-oxadiazoles and some related compounds. AB - Two novel series of 4-thiazolidinone derivatives, namely 2-substituted-3-([4-(4 methoxybenzoylamino)benzoyl]amino)-4-thiazolidinones (7a-e) and 2-[4-(4 methoxybenzoylamino)benzoylhydrazono]-3-alkyl-4-thiazolidinones (5a-c) together with 2-[4-(4-methoxybenzoylamino)phenyl]-5-(substituted phenyl)amino-1,3,4 oxadiazoles (6a-c) have been synthesised as title compounds. N(1)-[4-(4 methoxybenzoylamino)benzoyl]-N(2)-substituted methylene hydrazines (3a-e) and 1 [4-(4-methoxybenzoylamino)benzoyl]-4-substituted phenyl thiosemicarbazides (4a-f) were also prepared and used as intermediate to give the title compounds. All synthesised compounds were screened for their antimycobacterial activity against Mycobacterium tuberculosis H37Rv and antimicrobial activities against various bacteria and fungi. Compounds 7a and 7b were found as the most active derivatives demonstrating 90 and 98% inhibition of mycobacterial growth of M. tuberculosis H37Rv in the primary screen at 6.25 microg mL(-1), respectively. However, level II assay revealed that the MIC values were not less than 6.25 microg mL(-1). None of the compounds showed significant antimicrobial activity against the microorganisms used whereas 3a and 7a inhibited the growth of several bacteria and fungi. PMID- 11900863 TI - Synthesis, antiarrhythmic, and antihypertensive effects of novel 1-substituted pyrrolidin-2-one and pyrrolidine derivatives with adrenolytic activity. AB - A series of 1-substituted pyrrolidin-2-one and pyrrolidine derivatives were synthesised and tested for electrocardiographic, antiarrhythmic, and antihypertensive activity as well as for alpha(1)- and alpha(2)-adrenoceptors binding affinities. Among the newly synthesised derivatives several compounds with 3-(4-arylpiperazin-1-yl)propyl moiety displayed strong antiarrhythmic (7a 12a) and antihypertensive (7a-11a) activities. Compound 11a, 1-[2-acetoxy-3-[4-(2 methoxyphenyl)piperazin-1-yl]propyl]pyrrolidin-2-one, was the most potent in this series. The pharmacological results and binding studies suggest that their antiarrhythmic and hypotensive effects may be related to their alpha-adrenolytic properties, and that those properties depend on the presence of the 1 phenylpiperazine moiety with a methoxy- or chloro- substituent in the ortho position in the phenyl ring. PMID- 11900865 TI - Reactions of purines-containing butenolides with L-cysteine or N-acetyl-L cysteine as model biological nucleophiles: a potent mechanism-based inhibitor of ribonucleotide reductase caused apoptosis in breast carcinoma MCF7 cells. AB - Thiols are the most reactive nucleophilic reagents among the biological models investigated. The reactivity of butenolides 1a-c, 2-4, and 6-8 toward L-cysteine, a model biological nucleophile, was studied spectrophotometrically. The rates of the reactions were measured and correlated with antitumour activity of these molecules. N-Acetylcysteine addition product 5, resulting from the treatment of butenolide 4 with glutathione precursor, N-acetyl-L-cysteine, was isolated. Unlike purine-containing gamma-(Z)-ethylidene-2,3-dimethoxybutenolides 1a-c, 4, 6, and 7, adduct 5 and butenolides 10-12 did not exhibit inhibitory activity against murine leukemias (L1210 and P388), breast carcinoma (MCF7), and human T lymphoblasts (Molt4/C8 and CEM/0) cell lines. As such, the biological activity of purine-containing butenolides can be attributed to their adenine moiety as a recognition site as well as their reactivity towards the cysteine residues of functional proteins forming covalent bond via reverse Michael type addition. Adenine-containing phosphonothioanhydride derivative 8 was also synthesised. Its reaction with N-acetyl-L-cysteine produced N,S-diacetylcysteine and thiophosphonate 9. Compound 9 did not exhibit anticancer activity; yet its precursor 8 displayed the most pronounced inhibition on all the examined malignant tumour cell lines. In the presence of L-cysteine, cytotoxicity of 4 and 8 was decreased, whereas glutathione addition more influenced on the cytotoxicity of 8. It was found that adenine-containing phosphonothioanhydride 8 functions as a significant irreversible inactivator of the Escherichia coli ribonucleoside diphosphate reductase. After treatment of MCF7 cells with compound 8, fluorescence microscopy demonstrated the presence of nucleus shrinkage or segmentation. This apoptotic morphology, however, was not pronounced in the presence of glutathione or dithiotheritol. PMID- 11900866 TI - Local intersection volume: a new 3D descriptor applied to develop a 3D-QSAR pharmacophore model for benzodiazepine receptor ligands. AB - In this work, we have developed a new descriptor, named local intersection volume (LIV), in order to compose a 3D-QSAR pharmacophore model for benzodiazepine receptor ligands. The LIV can be classified as a 3D local shape descriptor in contraposition to the global shape descriptors. We have selected from the literature 49 non-benzodiazepine compounds as a training data set and the model was obtained and evaluated by genetic algorithms (GA) and partial least-squares (PLS) methods using LIVs as descriptors. The LIV 3D-QSAR model has a good predictive capacity according the cross-validation test by "leave-one-out" procedure (Q(2)=0.72). The developed model was compared to a comprehensive and extensive SAR pharmacophore model, recently proposed by Cook and co-workers, for benzodiazepine receptor ligands [J. Med. Chem. 43 (2000) 71]. It showed a relevant correlation with the pharmacophore groups pointed out in that work. Our LIV 3D-QSAR model was also able to predict affinity values for a series of nine compounds (test data set) that was not included into the training data set. PMID- 11900867 TI - Anticonvulsant and neurotoxicity evaluation of some 6-chlorobenzothiazolyl-2 thiosemicarbazones. AB - Ten 6-chlorobenzothiazolyl-2-thiosemicarbazones were synthesised and screened for anticonvulsant and neurotoxic properties. Most of the compounds showed anticonvulsant activity against both maximal electroshock seizure (MES) and subcutaneous pentylenetetrazole screens. Eight compounds have shown good protection in the rat p.o. MES test at 30 mg kg(-1). Compound 1 [4-(6 chlorobenzothiazol-2-yl)-1-(3-isatinimino)thiosemicarbazone] emerged as the most promising one with an ED(50) of 17.86 and 6.07 mg kg(-1) in mice i.p. and rat p.o., respectively. Compound 1 showed a weak ability to block the expression of fully kindled seizures. PMID- 11900868 TI - (2-Arylhydrazonomethyl)-substituted xanthones as antimycotics: synthesis and fungistatic activity against Candida species. AB - A series of arylhydrazones derived from various 6,8-diacetoxy- or 6,8-dihydroxy-9 oxo-9H-xanthene carboxaldehydes were synthesized and evaluated for their in vitro antifungal properties against two human pathogenic yeasts (Candida albicans and C. krusei) according to a diffusion method. The activity was strongly dependent from the position of the (1-arylhydrazinyl-2-ylidene)methyl chain in the xanthone molecular skeleton. Compounds having the nitrogen side chain in the 4-position, with a further halogen substitution on the terminal phenyl ring showed fungistatic effects. Within this series, the 4-fluorophenylhydrazinyl derivative 13g exhibited the highest activity, particularly against C. krusei, with a greater efficacy than that of econazole, used as reference. PMID- 11900870 TI - Synthesis and in vitro cytotoxic activity of pyrrolo[2,3-e]indole derivatives and a dihydro benzoindole analogue. AB - The synthesis of pyrrolo[2,3-e]indole derivatives with the structural characteristics of DNA bis- and mono-intercalators are described. A dihydro benzoindol analogue was also synthesised to elucidate the major structural requirements for cytotoxic activity. A biological evaluation of the test compounds was carried out in six different tumoral cell lines. The factors that affect the cytotoxic activity appear to be the substituents on the phenyl group, the presence of an amide group capable of strong interactions such as hydrogen bonding and solubility. PMID- 11900869 TI - Synthesis and anticancer and anti-HIV testing of some pyrazino[1,2 a]benzimidazole derivatives. AB - In this study, some 1-methylene-2,3-diaryl-1,2-dihydropyrazino[1,2 a]benzimidazole and some 1-(2-arylvinyl)-3-arylpyrazino[1,2-a]benzimidazole derivatives were synthesised. The structure elucidation of the compounds was performed by IR, 1H-NMR and MASS spectroscopic data and elemental analyses results. Anticancer and anti-HIV activities of the compounds were examined, however no anti-HIV activity was seen; highly notable anticancer activity was obtained. It was also observed that the compounds were more potent against leukaemia cell lines. PMID- 11900871 TI - Pyrrolo[2,1-c][1,2,4]triazines from 2-diazopyrroles: synthesis and antiproliferative activity. AB - Pyrrolo[2,1-c][1,2,4]triazines 4a-g were directly obtained from the reaction of 2 diazopyrroles 1a and b with the sodium salts of beta-diketones, beta carbonitriles, and beta-dinitriles. Only when the 2-diazopyrroles were coupled with ethyl cyanoacetate, it was possible to isolate, together with the pyrrolotriazines, the intermediate hydrazones 3 which, in turn, cyclised to the title ring system. Pyrrolotriazines 4a-e were evaluated for cytotoxic activity against a panel of 60 human cancer cell lines by the National Cancer Institute and some of them demonstrated inhibitory effects in the growth of a wide range of cancer cell lines generally at 10(-5) M level and in some cases at micromolar concentrations. PMID- 11900873 TI - Hereditary cancer: family history, diagnosis, molecular genetics, ecogenetics, and management strategies. AB - The translation of knowledge about hereditary breast cancer and its improved control, as well as prevention through prophylactic surgery, has been significantly accelerated through the veritable explosive discoveries in molecular genetics inclusive of BRCA1 and BRCA2 germline mutations. Needed however, among the physician community, medical geneticists, and genetic counselors, is a raised level of knowledge about hereditary breast cancer syndromes. Particular attention needs to be given to their extant genotypic and phenotypic heterogeneity, their natural history, and foremost, the requirement of a sufficiently detailed family history, with knowledge as to how to interpret its significance so that hereditary cancer syndrome can be diagnosed, should it, in fact, exist in the particular family. Collectively, surveillance and management programs can then be developed for the patient and his or her high-risk relatives. We believe very firmly that this knowledge needs to be extended to the individual patient(s), first- and second-degree relatives so that they can benefit from this knowledge. PMID- 11900874 TI - Population aspects of cancer genetics. AB - A number of relatively rare, high-risk genes have been identified which predispose to common cancers such as breast, colon, and melanoma. Although these are clearly important in the clinical setting, it is also relevant to discuss the impact of these genes at the population level and to contrast these with that which could be ascribed to more common genetic variants which only confer a modest increased risk of cancer. In this review, we examine inferences about the role of genetics in cancer from ecological studies of incidence patterns from a number of population-based studies of familial and attributable risk. The relationship between the genetic model (genotypic risk, allele frequency, mode of inheritance) and the expected impact in the population in terms of both attributable risk and familial risk is presented. The advantages and limitations of using cancer occurrence in twins to measure the genetic contribution to specific cancer sites is discussed. The potential role of lower-penetrance genes in the overall cancer burden may be significant but may pose significant problems in the public health arena. PMID- 11900876 TI - The genetics of the hereditary xeroderma pigmentosum syndrome. AB - All living organisms are constantly exposed to endogenous or exogenous agents that can cause damage to the genomic DNA, leading to the loss of stable genetic information. Fortunately, all cells are equipped with numerous classes of DNA repair pathways which are able to correct many kinds of DNA damage such as bulky adducts, oxidative lesions, single- and double-strand breaks and mismah. The importance of these DNA repair processes is attested by the existence of several rare but dramatic hereditary diseases caused by defects in one of their repair pathways. These diseases are usually associated with early onset of malignancies confirming the direct relationship between unrepaired DNA lesions, mutations or chromosomal modifications and cancer incidence. Among these hereditary diseases the UV-hypersensitive ones have been particularly well studied and the xeroderma pigmentosum (XP) is probably the best known syndrome up to now in terms of genetics and biochemistry. PMID- 11900875 TI - DNA mismatch repair defects: role in colorectal carcinogenesis. AB - The inactivation of the DNA mismah repair (MMR) system, which is associated with the predisposition to the hereditary non-polyposis colorectal cancer (HNPCC), has also been documented in nearly 20% of the sporadic colorectal cancers. These tumors are characterized by a high frequency of microsatellite instability (MSI(+) phenotype), resulting from the accumulation of small insertions or deletions that frequently arise during replication of these short repeated sequences. A germline mutation of one of the two major MMR genes (hMSH2 or hMLH1) is found in half to two-thirds of the patients with HNPCC, whereas in sporadic cases hypermethylation of the hMLH1 promoter is the major cause of the MMR defect. Germline mutations in hMSH6 are rare and rather confer predisposition to late-onset familial colorectal cancer, and frequent extracolonic tumors. Yet, the genetic background of a number of HNPCC patients remains unexplained, indicating that other genes participate in MMR and play important roles in cancer susceptibility. The tumor-suppressor genes that are potential targets for the MSI driven mutations because they contain hypermutable repeated sequences are likely to contribute to the etiology and tissue specificity of the MSI-associated carcinogenesis. Because the prognosis and the chemosensitivity of the MSI(+) colorectal tumors differ from those without instability, the determination of the MSI phenotype is expected to improve the clinical management of patients. This review gives an overview of various aspects of the biochemistry and genetics of the DNA mismah repair system, with particular emphasis in its role in colorectal carcinogenesis. PMID- 11900877 TI - Inheritable forms of medullary thyroid carcinoma. AB - Medullary thyroid carcinoma (MTC) arises from parafollicular or C cells of the thyroid that produce calcitonin. It accounts for 5-10% of all thyroid cancers. Hereditary MTC represents 20-30% of all MTCs. It can be transmitted with an autosomal dominant pattern, either as a single entity, familial MTC, or it can arise as part of a multiple endocrine neoplasia (MEN) syndrome type 2A or 2B. The identification of hereditary MTC has been facilitated in recent years by the direct analysis of the ret proto-oncogene. PMID- 11900878 TI - Genetic and environmental factors in cutaneous malignant melanoma. AB - Cutaneous malignant melanoma (CMM) is an interesting example of multifactorial disease, where both genetic and environmental factors are involved and interact. Major risk factors include a personal and familial history of melanoma, cutaneous and pigmentary characteristics, sun exposure and reactions to sun exposure. Phenotypic risk factors are likely to be genetically determined. Two high-risk melanoma susceptibility genes-CDKN2A and CDK4-have been identified to date, with a third gene p14(ARF) also being suspected of playing a role. Other high-risk genes are anticipated by the existence of 9p21-unlinked families. A low-risk melanoma-susceptibility gene-MC1R-has also been identified. Current studies aim to identify other susceptibility genes as well as to determine the respective contributions and interactions of the various genetic and environmental factors of CMM and associated phenotypes. PMID- 11900879 TI - The Li-Fraumeni syndrome. AB - Li-Fraumeni syndrome (LFS) has been the most common terminology used for the syndrome. It is a rare familial dominantly inherited cancer syndrome characterized by a wide spectrum of neoplasms occurring in children and young adults. The canonical definition of LFS includes a proband diagnosed with sarcoma before 45 years of age, a first-degree relative with cancer before this same age and another first- or second-degree relative in the lineage with any cancer before this age or sarcoma at any age. Multiple studies have reported p53 germline mutations in LFS families in various parts of the world. As in sporadic tumors, loss of heterozygosity leading to the inactivation of the wild-type allele by deletion or mutation is observed in LFS tumors. Cancer-risk in mutation carriers has been estimated to be 73% in males and nearly 100% in females, the difference almost entirely explained by breast cancer. The identification of germline p53 mutations in rare cancer-prone families has given rise to the medical, counseling, psychological and ethical problems. PMID- 11900880 TI - TP53: a key gene in human cancer. AB - TP53 is mutated in most types of human cancers and is one of the most popular genes in cancer research. The p53 protein is a sensor of multiple forms of genotoxic, oncogenic and non-genotoxic stress. It suppresses growth and controls survival of stressed cells, and as such, is the focal point of selection pressures in tissues exposed to carcinogens or to oncogenic changes. Thus, the clonal expansion of cells with mutations in TP53 may be seen as the result of a selection process intrinsic to the natural history of cancer. In this review, we discuss the nature of these various forms of selection pressure. We present a hypothesis to explain why TP53 is often mutated as either an early or a late event in cancer. Furthermore, we also summarise current knowledge on the molecular consequences of mutation for loss of wild-type protein function, dominant-negative activity, and a possible gain of oncogenic function. PMID- 11900881 TI - BRCA1 and BRCA2 in hereditary breast cancer. AB - The hereditary breast and ovarian cancer susceptibility genes, BRCA1 and BRCA2, have established roles in genome integrity maintenance and in the control of homologous recombination. Recent work has produced valuable insights into the mechanisms of action of the gene products. This review summarizes some of these advances, and attempts to place them in the context of known functions of the genes. PMID- 11900882 TI - Endocytosis of G protein-coupled receptors: roles of G protein-coupled receptor kinases and beta-arrestin proteins. AB - Sequestration of G protein-coupled receptors from the cell surface is a commonly observed phenomenon following agonist-stimulation. This process is now believed to be important for receptor resensitization as well as for signal transduction. Over the years, numerous studies have aimed at understanding the molecular mechanisms underlying internalization. Proteins such as the G protein-coupled receptor kinases (GRKs) and the beta-arrestins, which were initially characterized as desensitizing molecules, have been shown to be important regulators of the endocytic process. Recently, numerous interacting partners have been identified for each of these two classes of proteins. However, the details regarding the sequence of these interactions and the cross-talk between signaling pathways containing the different protein complexes are just beginning to be uncovered. In this review, we summarize these findings and discuss the role of GRKs and beta-arrestins, two families of key regulatory proteins that regulate G protein-coupled receptor endocytosis. PMID- 11900883 TI - The medullary dorsal reticular nucleus as a pronociceptive centre of the pain control system. AB - The endogenous pain control system has long been considered as engaged in pain depression through the commitment of multiple descending actions that reduce the response capacity of spinal dorsal horn nociceptive neurones. Such a pure inhibitory antinociceptive nature was lately questioned by the observation of pronociceptive effects from areas classically regarded as antinociceptive. The thereby raised hypothesis of a more versatile functional arrangement that dynamically adjusts the pain modulatory effect to multiple conditions by balancing several excitatory and inhibitory actions found strong support on the recent discovery of a medullary area particularly dedicated to pain facilitation. Lesioning the medullary dorsal reticular nucleus (DRt) depresses nociceptive responses to acute and inflammatory pain, whereas stimulation produces the inverse effect. The decrease in formalin-induced pain behaviour following DRt lesioning is accompanied by a decrease of spinal noxious-evoked c-fos neuronal activation. DRt blocking by lidocaine results in a decrease of the nociceptive activity of spinal dorsal horn neurones, whereas stimulation by glutamate has the opposite effect. A reciprocal disynaptic putative excitatory circuit that links the DRt and the spinal dorsal horn and conveys nociceptive input through the ascending branch was described, indicating that the DRt pain facilitating action is mediated by a reverberating spino-DRt circuit that promotes the enhancement of the response capacity of spinal neurones to noxious stimulation.The demonstration of a primary pronociceptive centre in the endogenous pain control system brings new important data to the emerging concept of pain modulation as a dynamic and flexible process that integrates nociceptive processing by balancing multiple excitatory and inhibitory actions as the way of adapting to the various unsteady pain determinants. PMID- 11900884 TI - Recovery of motor and language abilities after stroke: the contribution of functional imaging. AB - In recent years, functional imaging techniques, like functional magnetic resonance imaging (fMRI), positron emission tomography (PET), and transcranial magnetic stimulation (TMS), have shown that the improvement of motor and language function after ischemic stroke is accompanied by extensive reorganizational changes in the human cortex. To better understand these changes and to judge to what extent they could be responsible for clinical improvement, some basic principles of the organization of the motor and language system are discussed.Non invasive functional imaging can have only a limited contribution in determining which of the possible underlying neural mechanisms, as they are known from animal experiments, play a role in functional recovery. However, they make it possible to define the functional consequences of anatomical lesions in individual patients and to correlate these functional consequences in the motor and language system with the clinical deficit. They can be used to assess the influence on the cortical reorganization of established and newer physiotherapies, logopedics and medical intervention, and they could be a useful tool in determining prognosis. PMID- 11900886 TI - Control of human luteal steroidogenesis. AB - The human corpus luteum (CL) undergoes a dynamic cycle of differentiation, steroid hormone production and regression during the course of non-fertile cycles. In humans and other primates, luteal steroidogenesis is absolutely dependent on pituitary-derived LH. However, changes in LH and LH receptor expression do not explain the marked decline in progesterone production at the end of the luteal phase. Changes in the level of the steroidogenic acute regulatory protein (StAR), a gene whose expression is controlled by LH most likely account for the cyclic pattern of progesterone production. During the mid to-late luteal phase of a fertile cycle, chorionic gonadotropin (hCG) rescues the CL, overcoming the actions of the factors inducing luteolysis. Although the agents causing regression of the CL in a non-fertile cycle are not yet known, intra-luteal growth factors and cytokines that modify the action of LH probably contribute to the reduction of StAR expression and the subsequent fall in progesterone production. PMID- 11900888 TI - Oogenesis as a foundation for embryogenesis. AB - The majority of oocytes in postnatal ovaries are small, non-growing and reside in primordial follicles. They have to undergo a prolonged phase of growth and differentiation before nuclear and cytoplasmic maturation enables them to resume meiosis and undergo fertilization. A better knowledge of this phase of oogenesis is essential for understanding causes of oocyte pathology and optimizing methods for growing oocytes in vitro and for cryopreservation. There could also be spin off discoveries for contraceptive strategies and pharmacologically controlling oocyte maturation. During oocyte growth, a molecular programme for development is assembled for the timely expression of mRNAs, some of which are expressed throughout oogenesis while others are 'masked' until or after meiotic maturation. Masking and stability in storage are largely due to a truncated poly(A) tail, controlled by regulatory sequences on the 3'-untranslated region (UTR) of the mRNA. Most maternal RNAs are degraded early in cleavage, there being a narrow overlap between persisting maternal mRNAs and activation of the embryonic genome. Accumulation of RNAs and proteins are not, however, the only major changes taking place during oogenesis. Cytoplasmic organelles multiply and redistribute, and there are epigenetic modifications of DNA for monoallelic expression of imprinted genes. The granulosa cells are obligatory for they provide physical support, nutrients and mediate the regulatory influences of gonadotrophic hormones. On the other hand, the oocyte actively influences the growth and differentiation of its granulosa cells. Thus, healthy embryos reflect the quality of both the oocyte and the granulosa cells. PMID- 11900887 TI - Endometrial function: cell specific changes in the uterine environment. AB - The uterus undergoes dynamic changes during the cycle and these events are largely driven by ovarian steroids. However, in the presence of an embryo, an additional series of changes that are not otherwise observed predominate. The ability of the embryo to modulate the uterine environment is restricted to a specific time of the cycle which is termed the 'window of receptivity'. Changes that occur within this window of receptivity and immediately following implantation can be divided into three distinct phases. The first phase, regulated by estrogen and progesterone, is characterized primarily by changes in both the luminal and glandular epithelial cells in preparation for blastocyst apposition and attachment. If the action of progesterone is antagonized, these changes are inhibited and the uterus is maintained in a pre-receptive state. The second phase is in the further modulation of these steroids induced changes by embryonic signals. In the primate, infusion of chorionic gonadotropin in a manner that mimics blastocyst transit, results in the endoreplication and plaque formation in the luminal epithelium. The glandular epithelium responds by increasing transcriptional and post-translational modifications of secretory proteins and the stromal fibroblasts initiate their differentiation process into a decidual phenotype. The final phase is associated with trophoblast invasion and remodeling of the endometrial stromal compartment. The most dramatic effect is on the stromal fibroblasts, which in response to embryonic stimuli, differentiate into decidual cells, the major cell type of the gestational endometrium. Thus, during the window of receptivity, signals from the embryo can dramatically alter the morphological and functional characteristics of the uterine endometrium. We suggest that these changes are critical to ensure prolonged maintenance of endometrial function during gestation and facilitate trophoblast invasion. PMID- 11900889 TI - Disruption of germ cell-Sertoli cell interactions leads to spermatogenic defects. AB - In the aging human testis, partially regressed tubules contain Sertoli cells with an altered appearance and reduced numbers of germ cells. Investigating the effects of aging on Sertoli cell-germ cell interactions from Brown Norway rats, we have found that a selective breakdown in germ cell-Sertoli interactions could lead to severe reductions in male fertility. Previous studies have identified two specific inducible germ cell markers (a von Ebner's-like protein and the Huntington disease protein) and two specific inducible Sertoli cell markers (a sertonin receptor and a novel gene) that are expressed in Sertoli cell-germ cell cocultures and in vivo [Endocrinology 140 (1999a) 5754; J. Biol. Chem. 27 (1999b) 10737]. We find that germ cells from aged regressed testes are unable to respond to selective signals from Sertoli cells, although germ cells from aged normal sized testes respond well. Similarly, Sertoli cells from aged regressed testes fail to respond to certain signals from young germ cells. We propose that selective disruptions in communication between Sertoli cells and germ cells contribute to germ cell loss during aging. PMID- 11900890 TI - Gonadotropin therapy for the treatment of anovulation and for ovarian hyperstimulation for IVF. AB - Knowledge of the mechanisms of single dominant follicle selection has led to the development of a novel and effective ovulation induction regimen for anovulatory women; the step down protocol. This commences with a fixed high gonadotropin dose followed by several decremental steps. For some patients the initial dose is too high, risking ovarian hyperstimulation syndrome. A major improvement to this approach would, therefore, be the ability to use initial screening characteristics to assess the individual FSH threshold beforehand. For IVF treatment, interfering in the process of single dominant follicle selection in ovulatory women by late follicular phase administration of low doses of FSH may result in a significantly reduced duration of stimulation and amounts of exogenous FSH preparations used. Less monitoring would be required and chances for short-term complications or long term risks may be reduced. PMID- 11900891 TI - Germ cell transplantation. AB - Transplantation of germ cells leads to restoration of spermatogenesis from spermatogonial stem cells. The original description of germ cell transplantation in 1994 has led to new approaches to explore many basic aspects of spermatogonial physiology. Combining germ cell transplantation with culture and cryopreservation of spermatogonia opens new pathways for genetic engineering and conservation of livestock. In addition, autologous transfer of spermatogonia might be used as an approach for fertility preservation in oncological patients. This review summarises the history of germ cell transplantation and presents an outlook on future research on spermatogonial stem cells. PMID- 11900893 TI - Paracrine dialogue in implantation. AB - We know that the implantation process requires a functionally normal embryo at the blastocyst stage and a receptive endometrium, but also a communication link between them is needed. This paracrine dialogue between the embryo, endometrium and the corpus luteum are known to occur in ruminants and primates, more specifically endometrial-embryonic interactions have been reported in rodents and primates but not in humans. This process is a highly regulated mechanism and many molecules take part in this cross-talk. Here, we present updated information in humans on the embryonic regulation of endometrial epithelial molecules such as chemokines, adhesion and anti-adhesion molecules, and leptin during the apposition and adhesion phases of human implantation. PMID- 11900892 TI - Perinatal neuroendocrine regulation. Development of the circadian time-keeping system. AB - During gestation, the perinatal neuroendocrine axis keeps clock time. In primates, the suprachiasmatic nucleus (biological clock in mammals), shows oscillatory function by midgestation. There is evidence in rodents that the mother, during pregnancy, entrains the fetal suprachiasmatic nucleus (SCN) and newborn circadian rhythms. We are investigating the role of maternal melatonin as an entraining signal for the newborn circadian time-keeping system in the Cebus apella (New World non-human primate). Twenty-four hour rhythms of temperature and cortisol are present in the 4 days old C. apella newborn. Preliminary data suggests that inhibition of maternal melatonin by exposing pregnant females to constant light alters these rhythms. We have found binding sites for melatonin and expression of mRNA for Mel 1A receptor in hypothalamus, kidney and testis. These preliminary results suggest that maternal melatonin may play a role in relating the perinatal circadian time-keeping system to environmental signals. PMID- 11900894 TI - Strategies to elucidate the mechanism of excessive theca cell androgen production in PCOS. AB - Polycystic ovary syndrome (PCOS) is characterized by increased ovarian androgen production. We have developed a strategy to identify differentially transcribed genes in PCOS theca cells. Using long-term cultures of normal and PCOS theca cells, we are characterizing the complete repertoire of differentially transcribed genes, the common transcription factor mediators, and the signal transduction cascade(s) that are abnormal in PCOS theca cells. The latter will be interrogated for mutations/genetic variants linked to PCOS. PMID- 11900895 TI - Assessment of the in vitro and in vivo biological activities of the human follicle-stimulating isohormones. AB - Gonadotropins are synthesized and released in different molecular forms. In this article, we present evidence that the glycosylation variants of human pituitary FSH exhibit differential and divergent effects at the target cell level and that less sialylated, short-lived variants may exert significant effects in in vivo conditions. Less acidic/sialylated glycoforms (elution pH value 6.60-4.60 as disclosed by high resolution chromatofocusing of anterior glycoprotein extracts), induced higher cAMP release, estrogen production and tissue-type plasminogen activator (tPA) enzyme activity as well as cytochrome P450 aromatase and tPA mRNA expression in cultured rat granulosa cells than the more acidic analogs (pH<4.76). By contrast, the more acidic/sialylated glycoforms induced higher alpha-inhibin subunit mRNA expression than their less acidic counterparts. In cumulus enclosed oocytes isolated from mice ovaries, addition of less acidic isoforms induced resumption of meiosis more efficiently than the more acidic analogs. Interestingly, the least acidic isoform (pH>7.10) behave as a strong antagonist of several FSH-mediated effects. Assessment of the in vivo effects of the isoforms on granulosa cell proliferation in follicles from immature rats, revealed that short-lived isoforms were equally or even more efficient than their more acidic counterparts in maintaining granulosa cell proliferation when administered immediately after hypophysectomy. These results show that the naturally occurring human FSH isoforms may exhibit differential or even unique effects at the target cell level and that factors other than the metabolic clearance rate of the molecule (including receptor-binding affinity and capability of the ligand to activate its receptor and trigger intracellular signaling) also play an important role in determining the net in vivo effects of a particular FSH variant. PMID- 11900902 TI - Proceedings of the International Conference on Reproductive Competence: Pathophysiology and Therapeutic Interventions. Santiago, Chile, November 28-30, 2000. PMID- 11900896 TI - Intracytoplasmic sperm injection. AB - Intracytoplasmic sperm injection (ICSI) with ejaculated, epididymal or testicular spermatozoa was first successful in 1992 and has since become the widely accepted treatment for couples with severe male-factor infertility. The outcome of several thousands of ICSI cycles in terms of fertilization, embryo cleavage and implantation is similar to that for conventional in-vitro fertilization in couples with tubal or idiopathic infertility. To evaluate the important issue of safety of the new technique of ICSI, a prospective follow-up study of children born after ICSI was carried out. The aim was to compile data on karyotypes, congenital malformations, growth parameters and developmental milestones. Parents' agreement to genetic counseling was obtained, as well as prenatal diagnosis, followed by a physical examination of the children at 2 months, 1 and 2 years. Important outcome data to be examined comprise information on major and minor congenital malformations obtained prenatally or after birth, as well as on the further development of the children. PMID- 11900903 TI - Use of the most likely failure point method for risk estimation and risk uncertainty analysis. AB - The most likely failure point (MLFP) method, developed within the field of structural reliability analysis (where it is known as the FORM/SORM method) is a technique for estimating the risk (probability) that a calculated quantity Q exceeds a set limit Q(lim) when some or all of the inputs to the calculation are uncertain. It can be used as an efficient stand-alone method for this type of risk calculation. However, for application within the field of toxic hazards, it is proposed as a means for performing sensitivity analyses, possibly in parallel with a risk calculation carried out by conventional methods. The basis of the method is outlined and its use is demonstrated by means of an example calculation of the risk arising from an installation containing chlorine. The calculation uses, as a consequence model, commercial software for the prediction of dense gas transport. The risk estimate is shown to be acceptably close to that obtained by the Monte Carlo method. The use of a proposed screening procedure utilising the sensitivity formulas that the method provides, in order to identify the most significant uncertainties, is demonstrated. The identification of a single set of input values containing sufficient information to summarise (at least approximately) the entire risk analysis is considered to be an important feature of the method and is proposed as the basis of a means for assessing the validity of the consequence model. PMID- 11900905 TI - A model for evaluating physico-chemical substance properties required by consequence analysis models. AB - Modeling systems for analyzing the consequences of chemical emergencies require as input values a number of physico-chemical substance properties, commonly as a function of temperature at atmospheric pressure. This paper presents a mathematical model "CHEMIC", which can be used for evaluating such substance properties, assuming that six basic constant quantities are available (molecular weight, freezing or melting point, normal boiling point, critical temperature, critical pressure and critical volume). The model has been designed to yield reasonably accurate numerical predictions, while at the same time keeping the amount of input data to a minimum. The model is based on molecular theory or thermodynamics, together with empirical corrections. Mostly, model equations are based on the so-called law of corresponding states. The model evaluates substance properties as a function of temperature at atmospheric pressure. These include seven properties commonly required by consequence analysis and heavy gas dispersion modeling systems: vapor pressure, vapor and liquid densities, heat of vaporization, vapor and liquid viscosities and binary diffusion coefficient. The model predictions for vapor pressure, vapor and liquid densities and heat of vaporization have been evaluated by using the Clausius-Clapeyron equation. We have also compared the predictions of the CHEMIC model with those of the DATABANK database (developed by the AEA Technology, UK), which includes detailed semi empirical correlations. The computer program CHEMIC could be easily introduced into consequence analysis modeling systems in order to extend their performance to address a wider selection of substances. PMID- 11900904 TI - Characterization of PAHs in the atmosphere of carbon black manufacturing workplaces. AB - The objective of this study was set out to characterize the polycyclic aromatic hydrocarbon (PAH) content in the atmosphere of an oil furnace carbon black manufacturing plant located in southern Taiwan. A standard semi-volatile sampling train, the PS-1 sampler, was used to collect samples from eight areas, including the feedstock oil unloading, furnace, filtering/micro-pulverization, pelletizing, packaging, office/outside, office/inside, and boundary area, respectively. For each area, side-by-side static samples were collected simultaneously and a total of 16 samples were obtained. For each collected sample, the adsorbent-retained PAH content and the filter-retained PAH content were used directly to determine the concentrations of gaseous-phase PAHs and particle-bound PAHs, respectively. The gas chromatograph/mass spectrometer (GC/MS) technique was used for PAH analyses, and a total of 21 PAH species were determined. Results show the gaseous phase PAHs accounted for only 69.2% of the total PAH content for samples collected from the packaging area, which was significantly lower than those samples collected from the rest of seven areas (ranging from 96.3 to 99.7%). The result is not so surprising since the packaging area had the highest dust concentration due to the releasing of carbon black dusts during the packaging process. In this study, we further examine the contribution of gaseous-phase PAHs to the total benzo[a]pyrene equivalent (BaP(eq)) content from the health-risk assessment view of point. It can be found the contribution of gaseous-phase PAHs to the total BaP(eq) content (63.1%) was quite comparable to the corresponding contribution to the total PAH content for samples collected from the packaging area. However, a different trend can be found for samples collected from the other seven areas, where the contributions of gaseous-phase PAHs to the total BaP(eq) content (ranging from 67.7 to 93.4%) were lower than the corresponding contributions to the total PAH content. The above results can be explained by PAH homologues that contained in both gaseous-phase and particle-bound PAHs. It was found the gaseous-phase PAHs contained higher fractions of less carcinogenic low molecular weight PAH homologues, whereas particle-bound PAHs contained higher fractions of more carcinogenic high molecular weight PAH homologues. Considering the contributions of gaseous-phase PAHs to both total PAH content and total BaP(eq) content were well above 50% for the eight studied areas, it is concluded that both particle-bound and gaseous-phase PAHs should be included for assessing the exposures of carbon black workers. PMID- 11900906 TI - Selection of a representative set of chemical accidents from a complex data matrix for the development of environment-accident index. AB - Chemical accidents often lead to negative consequences for the environment. Preparedness and proper actions are, therefore, essential components in order to minimise environmental effects. To assist and facilitate this work, a proposed planning tool, the environment-accident index (EAI), was formulated by Scott [J. Hazard. Mater. 61 (1998) 305]. As a result of a first validation of the index, based on 21 chemical accidents, the database was complemented with 42 additional accidents covering a broader spectrum of chemicals. The additional accidents were collected by means of an inquiry and their environmental consequences are, so far, unknown. The collected data had an overrepresentation of accidents involving petroleum products (69%). Because of the overrepresentation of this group of chemicals in the material, the data was skewed with respect to chemical properties. Since the model should be valid for a variety of chemical accidents, a method was needed which enabled a proper and unbiased selection of a representative subset of accidents to be used in development and validation of the model. For this purpose, the possibility to use multivariate data analysis in combination with statistical design was investigated. The result showed the feasibility of this method in the selection of a representative subset from a complex and skewed large dataset. Within the new dataset, 53% were accidents involving petroleum products and 47% involved other chemicals. The selected accidents will be used in further work to evaluate the environmental consequences, for model development and model validation. PMID- 11900907 TI - Vitrification of fly ash from municipal solid waste incinerator. AB - Fly ash from municipal solid wastes (MSW) incinerators in Korea contains a large amount of toxic materials and requires pertinent treatments. However, since fly ash in Korea has a high chlorine concentration, it is difficult to apply cementation and chemical treatment techniques. In this study, we report the vitrification of fly ash along with the properties of the glasses and leaching characteristics of heavy metal ions. Fly ash can be vitrified by melting at 1500 degrees C for 30 min with the addition of >5 wt.% of SiO2. Glasses showed Vickers hardness of 4000-5000 MPa, bending strength of 60-90 MPa and indentation fracture toughness of approximately 0.9 MPa m(1/2). Glasses also showed the excellent resistance against leaching of heavy metal ions with Cd2+ <0.04 ppm, Cr3+ <0.02 ppm, Cu2+ <0.04 ppm and Pb2+ <0.2 ppm. These results indicate that the vitrification technique is effective for the stabilization and recycling of toxic incinerator fly ash. PMID- 11900909 TI - Solubility enhancement of PCDD/F in the presence of dissolved humic matter. AB - From previous studies, we, the authors collected and arranged the octanol-water partition coefficients (K(ow)), water solubility (S0), and dissolved humic matter (DHM)-water partition coefficients (K(oc)) for 95 organic compounds, and presented the correlations between each physical property. The K(oc) and K(ow) of dioxins estimated were significantly increased while S0 was decreased on increasing the chlorine number. In the presence of DHM, solubility enhancement (Sw/S0, Sw is the actual solubility in the presence of DHM) in highly chlorinated PCDD/F such as HpCDDs and OCDD is higher than that in low chlorinated ones. It means that dioxins abundant wastes (fly ash) should not be codisposed with organic abundant wastes (sewage sludge, food waste or bottom ash, etc.) to minimize the leachability of dioxins. PMID- 11900908 TI - Use of hop (Humulus lupulus) agricultural by-products for the reduction of aqueous lead(II) environmental health hazards. AB - The agricultural by-products of the hop plant (Humulus lupulus L.) were investigated to determine their potential for use in the removal of heavy lead(II) ions from contaminated aqueous solutions. Separate batch laboratory experiments were performed to establish the optimal binding pH, time exposures, and capacity of the metal adsorption for lead(II) ions by dried and ground hop leaves and stems biomass. Results from these studies have shown a pH dependent binding trend from pH 2-6, with optimum binding occurring around pH 5.0. Time dependency experiments showed a rapid adsorption of lead(II) ions within the first 5 min of contact. Binding capacity experiments demonstrated that 74.2mg of lead(II) were bound per gram of leaf biomass. Similarly overall capacity was seen for the leaves and stems. Desorption of 99% of the bound lead(II) ions was achieved by exposing the metal laden biomass to 0.5M sodium citrate. Further experiments were performed with silica-immobilized hop tissues to determine the lead(II) binding ability under flow conditions. Comparison studies were performed with ion-exchange resins to evaluate the binding ability and to gain further insight into the metal binding mechanism. X-ray absorption spectroscopy experiments were also utilized to gain further insight into the possible lead(II) binding mechanism by the hop plant tissue. Results from these studies indicate that carboxyl ligands are involved in the binding of lead(II) from aqueous solution. These findings show that the use of hop agricultural waste products may be a viable alternative, for the removal and recovery of aqueous lead(II) ions from contaminated waters. PMID- 11900910 TI - Simultaneous control of acid gases and PAHs using a spray dryer combined with a fabric filter using different additives. AB - The purpose of this research was to simultaneously evaluate the removal efficiency of acid gases and PAHs from the flue gas emitted by a laboratory incinerator. This flue gas contained dust, acid gases, organics and heavy metals. A spray dryer combined with a fabric filter was used as the air pollution control device (APCD) in this study. The operating conditions investigated included different feedstock additives (polyvinyl chloride (PVC) and NaCl) and spray dryer additives (SiO2, CaCl2 and NaHCO3). The removal efficiency for SO2 could be enhanced by adding inorganic additives, such as SiO2, CaCl2 and NaHCO3. The presence of PVC in the incinerator feedstock also increased the removal efficiency of SO2in the spray dryer. The improved removal of PAHs could be attributed to the addition of feedstock additives (PVC and NaCl) and spray dryer additives (SiO2, CaCl2 and NaHCO3). PMID- 11900911 TI - Co-oxidation of p-hydroxybenzoic acid and atrazine by the Fenton's like system Fe(III)/H2O2. AB - The system Fe(III)/H2O2 has been used to oxidise an aqueous solution of p hydroxybenzoic acid (pHB) in the absence of light. In the process, typical operating variables such as reagent concentration exert a positive influence in the pHB degradation rate. Optimum pH has been found to be around 3. The kinetic study suggests that the mechanism involved in this system differs to some extent from that reported for the classic Fenton's chemistry in pure water. Thus, formation of a complex Fe(III)-pHB seems to be a key step to initiate the oxidising mechanism. Stoichiometric measurements of the H2O2 consumption per mole of pHB degraded indicate a possible reduction of complexed Fe(III). Simultaneous oxidation of pHB (and other similar compounds such as tyrosol (Ty) or p-coumaric acid (pCu)) and atrazine have shown a synergistic effect of the first substance to remove the pesticide. PMID- 11900912 TI - The effect of storage in an inert atmosphere on the release of inorganic constituents during intermittent wetting of a cement-based material. AB - Monolithic waste materials (e.g. Portland cement treated wastes) in many field scenarios do not remain continuously saturated, but experience intermittent wetting interspersed with periods of storage in an unsaturated environment. During storage, the matrix may loss moisture to the environment, promoting precipitation or redistribution of species. In addition, the matrix may react with the surrounding atmosphere through carbonation or oxidation. Upon subsequent leaching, changes in the chemical and physical composition incurred over the storage interval can influence the release of inorganic species. Current assessment approaches, which use continuous leaching data to project release over some assessment interval, do not allow for changes in leachability resulting from intermittent wetting and storage. Thus, this study evaluates the effect of storage intervals in an inert atmosphere on subsequent release of inorganic species from a synthetic Portland cement matrix. Tank leaching in deionized water was interspersed with storage at three relative humidity (RH) levels (nominally 0, 50 and 100% RH) in a 100% nitrogen atmosphere. Leaching data from the three intermittent wetting cases were compared to continuous leaching for the release of structural species (Ca, OH), highly soluble species (Na, K, Cl) and pH dependent species (As, Cd, Pb). The RH of storage environment, which acted as a boundary condition for the drying process, influenced the precipitation of species within dried pores and relaxation of pH and concentration gradients within water-filled regions. Gradient relaxation resulted from continued mass transport within saturated pores over the storage interval and was most evident when storage was conducted at 98% RH. However, when storage RH promoted drying of the matrix, the effect of gradient relaxation was balanced by precipitation. When release was normalized to total leaching time, relaxation of concentration gradients of highly soluble species resulted in greater cumulative release for the intermittently wetted cases than in the case of continuous leaching. The release of pH-dependent constituents was controlled by relaxation of the pH gradient and species solubility as a function of local pore water pH. Application of a current assessment protocol to estimate intermittent wetting release resulted in either over or underestimation of actual cumulative release, depending on the nature of the constituent of interest. These results imply that long-term constituent release from Portland cement-based waste forms should not be made by simple correction of saturated release assessments because alterations to the matrix leachability induced by the storage environment need to be considered. PMID- 11900913 TI - Adsorption of zinc by activated carbons prepared from solvent extracted olive pulp. AB - Activated carbons have been prepared by a two-step physical activation with steam at different burn-off levels to study the porosity development and its effect in zinc adsorption from aqueous solutions. The main material used was the residual from the extraction with solvent of the kernel-oil [solvent extracted olive pulp (SEOP)]. Olive, apricot and peach stone have been also used as different precursors. The products were characterized by N2 at 77K adsorption, Hg porosimetry and iodine number determination. The influence of surface complexes and pH has been investigated in an attempt to elucidate the adsorption phenomena. The effect of different treatments [demineralization with H2SO4 and oxidation with (NH4)2S2O8] was also evaluated for the adsorption of zinc species. Both basic and acidic carbons, originated from SEOP, show remarkable adsorption ability at solution pH=7. Their adsorption ability mainly depends on the content and nature of functional surface groups, the ash content of the precursors and the pH of the solution. These activated carbons were proved to be efficient adsorbents for the removal of water pollutants and contaminants. PMID- 11900914 TI - Effects from different types of construction refuse in the soil on electrodialytic remediation. AB - At abandoned industrial sites some of the previous buildings are often left behind. If the soil at such site is polluted with heavy metals and is to be remediated by an electrochemical method, the construction refuse within the soil matrix will influence the remediation action. The influence of different sorts of construction refuse on electrodialytic soil remediation was investigated in laboratory cells. An insulator, a stone, resulted in an uneven Cu removal in the close vicinity of the stone itself. An electric conductive screw disturbed the Cu removal due to the redox reactions occurring at the surface of the screw causing pH changes in the soil. Two types of refuse with ionic conducting properties were placed within the test cell, a piece of brick and concrete. The brick did not influence the Cu removal from the soil to a high extent, but it was seen that during the remediation the Cu concentration in the brick itself increased. In the case of concrete the Cu mobilized from the soil was simply found to adsorb very strongly to the concrete and thus the Cu could not be removed from the soil and the concrete as a whole. Furthermore, the removal of Cu in the soil next to the concrete was quite poor. It is very important to be aware of the presence of construction refuse at such sites when planning an electrochemical remediation action. All the refuse types investigated here influenced the Cu removal negatively compared to the reference experiment. PMID- 11900915 TI - An investigation into the sorption of heavy metals from wastewaters by polyacrylamide-grafted iron(III) oxide. AB - An adsorbent for heavy metals was synthesized by introducing carboxylate functional group into polyacrylamide-grafted hydrous iron(III) oxide. The product exhibits a very high adsorption potential for Pb(II), Hg(II) and Cd(II). The removal of metal ions by adsorption on adsorbent has been found to be contact time, concentration, pH and temperature dependent. The process follows first order reversible kinetics. The intraparticle diffusion of metal ions through pores in the adsorbent was shown to be the main rate-limiting step. The optimum pH range for the removal of metal ions was found to be 5.0-6.0. The thermodynamic parameters such as free energy change, enthalpy change and entropy change have been calculated to predict the nature of adsorption. The adsorption data were fitted using the Langmuir equation and maximum adsorption for each metal was estimated using their respective Langmuir equation constants. The method was applied for synthetic wastewaters. NaCl regeneration has been tried for several cycles with a view to recover the adsorbed metal ions and also to restore the sorbent to its original state. PMID- 11900916 TI - High temperature oxidation of C2Cl4/CH4 mixtures. AB - Experiments on high temperature oxidation of multi-chlorinated hydrocarbons, tetrachloroethylene (C2Cl4), with hydrocarbon fuels, CH4, were performed in a 15 mm i.d. tubular flow reactor. Temperatures ranged from 700 to 850 degrees C, with the average residence time in the range from 0.3 to 1.5s. Three equivalence ratios, phi=0.87 (fuel-lean (FL)), phi=1 (stoichiometry (S)), and phi=1.3 (fuel rich (FR)), were studied. The global Arrhenius equations for the decomposition of C(2)Cl(4) for each reactant set ratio are: k(lean)=5.77 x 10(15) exp(-30447/RT), k(stoi)=5.15 x 10(15) exp(-30421/RT), and k(rich)=6.32 x 10(14) exp(-28879/RT). The important reactions for destruction of parent C2Cl4 include: C2Cl4 --> C2Cl3 + Cl, C2Cl4 + H--> C2Cl3 + HCl and C2Cl4 + H --> C2HCl3 + Cl. The resulting reactant loss, and intermediate and final product profiles were determined. C2HCl3, C2Cl2, CO, CO2 and HCl are the major products for the reaction of C2Cl4/CH4/O2 mixtures for these three reaction systems. Minor intermediates include C2H3Cl, C2HCl, COCl2, CH3CHCl2, C2H4, C2H6, CCl2CHCH3 , trans-CHClCHCl, cis-CHClCHCl, trans-ClHC=CClCH(3), C6H6, and Cl2. The experimental data showed that as the oxygen concentration increased, the temperature needed to detect the resulting products decreased. PMID- 11900917 TI - Hydrometallurgical recovery of zinc and lead from electric arc furnace dust using mononitrilotriacetate anion and hexahydrated ferric chloride. AB - The purpose of this work was to study the feasibility at laboratory-scale of a new hydrometallurgical process for treating electric arc furnace dusts (EAFD). The proposed process is intended to extract zinc and lead from EAFD without destroying the iron oxides matrix. So, this material can be recycled by the steel industry. Independently of the origin of the samples, major mineralogical forms present in these wastes are Fe3O4, ZnO, ZnFe2O4 and PbOHCl. The proposed process consists of a hydrometallurgical treatment of wastes based on selective leaching of zinc and lead. Initially, a leaching is carried out utilizing a chelating agent, nitrilotriacetate anion (NTA3-), as the protonated form HNTA2-. Treatment of five EAFD samples for an hour at room temperature with a molar solution of reagent results in total leaching of the ZnO. In all cases the solubilized iron does not exceed 3 wt.%. The recovery of zinc and lead is performed by precipitation of metallic sulfides with a solution of Na2S4 sodium tetrasulfide 2M. These metallic sulfides can be used as metallurgical raw materials and the chelating reagent can be reused in the process after pH adjustment. The results of the normalized leaching test AFNOR X31-210 conducted on the leaching residues, shows that all the samples meet acceptance thresholds for hazardous wastes landfill. However, the residues contain a considerable amount of zinc as ZnFe2O4. The extraction of the zinc element requires the destruction of the ferrite structure. In this process, ZnFe2O4 is treated by FeCl3.6H2O. The reaction consists in a particle O2-/Cl- exchange allowing the recovery of zinc as ZnCl2 and iron as hematite Fe2O3. The separation of these products is accomplished by simple aqueous leaching. All of the zinc is extracted in a 8h treatment at 150 degrees C with a molar ratio FeCl3.6H2O/ZnFe2O4 equal to 10. Ultimate solid residues, which have been concentrated in iron, can be oriented towards the steel industry. PMID- 11900918 TI - Air monitoring of a coal tar cleanup using a mobile TAGA LPCI-MS/MS. AB - Real-time detection of air toxics is becoming increasingly important in understanding the sources and constituents of air pollution. The detection of low levels of air contaminants requires reliable sampling and calibration techniques, as well as sophisticated analytical instrumentation. Recently, a new low pressure chemical ionization (LPCI) source was developed for ambient air monitoring of benzene, toluene, xylene (BTX) and gas-phase polycyclic aromatic hydrocarbons (PAH) in real-time. This ion source in conjunction with a triple quadrupole (Q1, Q2, Q3) mass spectrometer (trace atmospheric gas analyzer (TAGA IIe)) has been proven highly useful for measuring selected air pollutants. The ion chemistry under LPCI conditions involves charge transfer reactions yielding parent ions which are selected in the first quadrupole, Q1, dissociated in the second quadrupole, Q2, and the resultant daughter ions are then identified by the third quadrupole, Q3. Monitoring of specific parent/daughter (P/D) ion pairs is used to measure concentrations of BTX and selected PAH. The response of the TAGA IIe is characterized through multi-point calibration curves. Detection limits (DL) as low as 0.5 microg/m(3) for BTX and PAH were accomplished by optimizing various TAGA IIe operating parameters. This unique method was applied in November 1999 to monitor emissions released during the cleanup of a historical coal tar site in Kingston, Ontario. This information was used by local officials for enhancing abatement activities or in some cases temporarily halting the excavation when levels of air toxics were higher than allowable provincial guidelines. PMID- 11900919 TI - The catalytic oxidation of aromatic hydrocarbons over supported metal oxide. AB - The catalytic activity of metals (Cu, Mn, Fe, V, Mo, Co, Ni, Zn)/gamma-Al2O3 was investigated to bring about the complete oxidation of benzene, toluene and xylene (BTX). Among them, Cu/gamma-Al2O3 was found to be the most promising catalyst based on activity. X-ray diffraction (XRD), Brunauer Emmett Teller method (BET), electron probe X-ray micro analysis (EPMA) and temperature programmed reduction (TPR) by H2 were used to characterize a series of supported copper catalysts. Increasing the calcination temperature resulted in decreasing the specific surface areas of catalysts and, subsequently, the catalytic activity. Copper loadings on gamma-Al2O3 had a great effect on catalytic activity, and 5 wt.% Cu/gamma-Al2O3 catalyst was observed to be the most active, which might be contributed to the well-dispersed copper surface phase. Using TiO2 (anatase), TiO2 (rutile), SiO2 (I) and SiO2 (II) as support instead of gamma-Al2O3, the activity sequence of 5 wt.% Cu with respect to the support was gamma-Al2O3 > TiO2 (rutile) > TiO2 (anatase)>SiO2 (I) > SiO2 (II), and this appeared to be correlated with the distribution of copper on support rather than with the specific surface area of the catalyst. The smaller particle size of copper, due to its high dispersion on support, had a positive effect on catalytic activity. The activity of 5 wt.% Cu/gamma-Al2O3 with respect to the VOC molecule was observed to follow this sequence: toluene > xylene > benzene. Increasing the reactant concentration exerted an inhibiting effect on the catalytic activity. PMID- 11900920 TI - Speciation of copper in the incineration fly ash of a municipal solid waste. AB - The speciation of copper and zinc in the incineration fly ash of a municipal solid waste in Taiwan was investigated in the present work. By the least-squares fitted X-ray absorption near edge structural (XANES) spectroscopy, we found that CuCO3, CuOH2, and CuO (fractions of 0.09, 0.39 and 0.51, respectively) were the main copper species in the fly ash. Quantitative analysis of the extended X-ray absorption fine structural (EXAFS) spectra indicated that the bond distance of Cu O in the fly ash was 1.96 A with a coordination number (CN) of 3.9 in the first shell of copper. In the second shell, the bond distance and CN of Cu-(O)-Cu were 2.91 A and 2.7, respectively. In addition, speciation of Zn was also examined in the same X-ray absorption energy (8780-9970 eV). The bond distance of Zn-O and Zn O-Zn were 1.97 and 2.94 A, respectively. However, the Zn-O-Cu structure was not found because of the physically unreasonable sigma(2) (Debye-Waller factor) values in the EXAFS data fitting process. PMID- 11900923 TI - Natural transmission routes of Neospora caninum between farm dogs and cattle. AB - Twelve dairy herds with evidence of post-natal infection with Neospora caninum were compared with 21 control herds with no evidence of post-natal infection. On the former farms, dogs consumed placenta or licked uterine discharge in 75 and 67% of the farms, respectively, while on control farms these activities occurred in 38 and 24% of the farms, respectively. On all control farms and all but three post-natally infected farms the dogs were fed colostrum or milk. Defecation of dogs on the feeding alley was observed in 92% of the post-natally infected farms and in 24% of the control farms. The same trend was observed for defecation of dogs in grass silage, in 75% of the post-natally infected farms and in 19% of the control farms; and in corn silage, in 50% of the post-natally infected farms and in 10% of the control farms. Consumption of placenta, material of aborted foetuses or uterine discharge in combination with defecation on the feeding alley, storage of grass or corn silage was observed in 19% of the control farms and in 75% of the post-natally infected farms. This study supports the hypothesis that farm dogs may become infected by foetal fluids or placental material of infected cattle, and may subsequently cause a post-natal infection of cattle in the herd by shedding oocysts. PMID- 11900922 TI - Point source exposure of cattle to Neospora caninum consistent with periods of common housing and feeding and related to the introduction of a dog. AB - Eight dairy herds with evidence of post-natal transmission of Neospora caninum were used to test the hypothesis of a point source exposure by a retrospective analysis of the housing and feeding of infected age-groups. The first N. caninum associated abortion or birth of N. caninum-seropositive offspring from the post natally infected age-group was considered as the first indication of the infection. In seven of the eight dairy herds, a point source exposure to N. caninum of the infected age-groups was found during a limited period of common housing and feeding. In all herds studied, the analysis indicated that the cattle had been infected shortly before the first abortions occurred. In all, except one herd, the post-natal infection was more directly related to housing than to feeding. Therefore, it appeared that the feed was contaminated in the feeding alley. In one herd, the total mixed ration was found to be the probable path of infection. In all farms studied, a new dog (young, adult dog or litter) had been introduced within a period of 1.5 years prior to the first indication of N. caninum infection in the cattle. As there was evidence in all herds of vertical transmission of neosporosis for years, it is hypothesized that the newly introduced dog was infected with N. caninum by materials from already infected cattle and subsequently transmitted the infection to other cattle by shedding of oocysts. PMID- 11900924 TI - Theileria buffeli infection of a Michigan cow confirmed by small subunit ribosomal RNA gene analysis. AB - Theileria buffeli, generally a benign parasite of cattle, has been reported in animals from Texas, Missouri and North Carolina. To date, there have been no reports of the parasite in cattle residing in northern portions of the US. An 8 year-old cow (Maine Anjou x Angus cross-bred) in Michigan presented with hemoglobinuria and a packed cell volume of 9. Blood films stained with Giemsa showed numerous intraerythrocytic parasites morphologically consistent with T. buffeli. The parasite was confirmed to be T. buffeli by SSU rRNA gene sequence analysis (SSU rRNA sequence, Type A). This represents the first report of this parasite in an animal in Michigan. PMID- 11900925 TI - Comparative pathogenicity of three genetically distinct Trypanosoma congolense types in inbred Balb/c mice. AB - Inbred Balb/c mice were infected with three clones of Trypanosoma congolense (Sam.28.1, Dind.3.1 and K60.1A) corresponding, respectively, to the three genetically distinct types (savannah, forest and kilifi) defined within this species, for the purpose of comparing their pathogenicity for a better understanding of the epidemiology of African trypanosomosis. Another clone of savannah type, IL 3000, was also tested simultaneously to study a probable strain variation. Both the clones of savannah type were found of extreme virulence with loss of appetite, rough hair, rapid respiration, lethargy, and all mice died within a week. Parasitaemias evolved rapidly to the first peak by day 3-5 post inoculation without any remission and the course of disease was correlated positively with the prepatent period. The clones of the forest type and the kilifi type were of low virulence with chronic infection and symptoms progressively less patent throughout the infection; only one mouse died in each experimental group. PMID- 11900926 TI - Production and characterization of a monoclonal antibody against recombinant fatty acid binding protein of Fasciola gigantica. AB - In Fasciola parasites fatty acid binding proteins (FABPs) are the carrier proteins that help in the uptake of fatty acids from the hosts' fluids. Attempts have been made to utilize both native and recombinant FABP (rFABP) for immunodiagnosis and vaccine development for fasciolosis. In this study, we have produced a number of monoclonal antibodies (MoAbs) against rFABP of Fasciola gigantica. These MoAbs were initially screened against rFABP by ELISA and then tested for their specificities by immunoblotting. Five stable clones were selected and characterized further: four of them were of the isotype IgG(1) while one clone was IgG(2a). All the MoAbs reacted with rFABP which has a molecular weight (MW) of 20 kD and with at least two isoforms of native proteins at MW 14.5 kD that were present in the tegumental antigen (TA) and crude worm extracts, and the excretion-secretion materials. Immunoperoxidase staining of frozen sections of adult parasites by using these MoAbs as primary antibodies indicated that FABP were present in high concentration in the parenchymal cells and reproductive tissues, in low concentration in the tegument and caecal epithelium. All MoAbs cross-reacted with a 14.5 kD antigen present in the whole body (WB) extract of Schistosoma mansoni, while no cross-reactivities were detected with antigens from Eurytrema pancreaticum and Paramphistomum spp. PMID- 11900927 TI - Cooperia punctata trickle infections: parasitological parameters and evaluation of a Cooperia recombinant 14.2 kDa protein ELISA for estimating cumulative exposure of calves. AB - Three groups of four calves each were trickle infected with three different levels of Cooperia punctata: 310 (group A), 1250 (group B) and 5000 (group C) third stage infective larvae (L3) twice a week over a 17-week period. Group D was the non-infected control group. Parasitological parameters as faecal egg counts (epg), worm burdens, size of worms and number of eggs per female were collected and the differences between the groups compared. Serological analyses were also conducted to investigate the efficiency of a recombinant C. oncophora CoES 14.2kDa protein in an ELISA to detect C. punctata antibodies. Group C had higher faecal egg counts until week 9 when the values decreased to those in group B. Mean faecal egg counts in group A were always lower than in the two other infected groups. The worm burdens were highest in group C, and lowest in group A, although the number of worms as a percentage of total larval intake was higher for the lower group. The mean length of the worms was shorter and the number of eggs per female lower for group C than for both other groups. ELISA using the CoES 14.2kDa proved to be efficient in measuring C. punctata antibodies. For group C it took 4 weeks to get increased levels of antibodies and this was one and 2 months more for groups B and A, respectively. Overall, there was a congruent relation between C. punctata antibodies and the cumulative exposure to the three different levels of trickle infections. PMID- 11900928 TI - Epidemiological studies on gastrointestinal helminths of dromedary (Camelus dromedarius) in semi-arid lands of eastern Ethiopia. AB - A total of 752 dromedaries were examined and 75% were found to be harbouring nematode eggs. The mean EPG was 1831 and the range was from 100 to 21,200. The prevalence rates in the four-age groups examined were 59.6% (3-7 years), 72.4% (8 12 years), 76.1% (13-17 years) and 83.9% (18-22 years). The prevalence rate for females and males were 77.6 and 64.8%, respectively and for long dry, short rainy, short dry and long rainy seasons were 66, 80, 69 and 82.6%, respectively. The mean EPG of faeces was significantly (P<0.01) higher for older animals compared to other group of younger animals (3-7 years), for females compared to males, and for rainy compared to dry season. Sixteen dromedary gastrointestinal organs were used for identification and counts of helminths. Among the adult worms identified, from the abomasum, Haemonchus longistipes had a 94% prevalence rate. From the small intestine Trichostrongylus colubriformis, Trichostrongylus probolurus, Impalaia tuberculata and Strongyloides papillosus were identified with prevalence rates of 75, 25, 63 and 20%, respectively. Moreover, cestodes such as Moniezia benedeni, Moniezia expansa, Avitellina spp. and Stilesia globipunctata with prevalence rates of 31, 13, 25 and 19%, respectively, were identified. I. tuberculata was identified for the first time in this country from a dromedary.The pathological lesions were more pronounced in higher infestations. Infestation level over approximately 1000 of H. longistipes and 15,000 in mixed infection of T. colubriformis and I. tuberculata, caused gross lesions of ulcerated and hyperaemic mucosa, and the odour of the fluid were fetid. The microscopic lesions observed were sloughing of epithelium, necrosis of glands, atrophy and loss of villi, haemorrhages and cellular infiltration mainly of eosinophiles and lymphocytes. PMID- 11900929 TI - Immunization of sheep against Fasciola gigantica with glutathione S-transferase. AB - Glutathione S-transferase in Fasciola gigantica (FgGST) was isolated by affinity chromatography, by which highly purified enzyme was obtained. FgGST on the SDS PAGE showed three protein bands ranging 24.5-26.5kDa. Glutathione S-transferase (GST) activity was determined by HABIG method. FgGST was evaluated as vaccine alone or in combination with either aluminum hydroxide or saponin in sheep against F. gigantica infection. ELISA was used for detection of anti-FgGST IgG. After vaccination, all sheep were challenged with 120 metacercaria of F. gigantica. The results indicated that anti-GST IgG was not elevated after challenge. All sheep were slaughtered 24-26 weeks after challenge. The results indicated that, although after second vaccination, antibody titers rose markedly in GST-Al(OH)(3) and GST-saponin groups, but declined 4 weeks after challenge. No correlation between anti-GST IgG titers and protection was observed. The highest fluke burden reduction was observed in the group vaccinated with GST-saponin (32%), but this reduction was not statistically significant in comparison with the control group. PMID- 11900930 TI - Echinococcosis/hydatidosis in western Iran. AB - In the present study, 115 stray dogs (56 males and 59 females, mixed breed), 86 golden jackal (Canis aureus, 42 males and 44 females), 60 red foxes (Vulpes vulpes, 33 males and 27 females), and three female wolves (Canis lupus) were examined for Echinococcus granulosus infection, as well as, 32,898 sheep, 10,691 goats, 15,779 cattle and 659 buffaloes for hydatid infection from five provinces in western Iran during 3 years (1997-2000). Meanwhile fertility rates of different types and forms of cysts isolated from infected animals and the viability of protoscolices were also determined. Results indicated that 19.1% of the dogs, 2.3% of the golden jackals and 5% of the red foxes were infected with Echinococcus granulosus. 11.1% of the sheep, 6.3% of the goats, 16.4% of the cattle and 12.4% of the buffaloes were also found to be infected with hydatid cyst. The cysts isolated from liver and lungs of the sheep show higher fertility rate than the cysts of liver and lungs of goats, cattle and buffaloes. PMID- 11900932 TI - Epidemiological data of patients hospitalized with burns and other traumas in some cities in the southeast of Brazil from 1991 to 1997. AB - This retrospective analysis of burn patients and victims of other forms of trauma from Ribeirao Preto and nearby cities admitted to hospitals in the city of Ribeirao Preto, Sao Paulo, Brazil, was carried out to determine the frequency of injuries of all types in order to identify the extent of the problem of burns relative to other forms of trauma. Data concerning 921 patients with burns and 60,344 patients with other traumatic injuries hospitalized during the period from 1991 to 1997 are described. Burns corresponded to 1.5% of the total number of traumatic injuries. When data are reported as absolute numbers or as incidence rate of hospitalized burn patients, burns were two times more frequent among men in most age groups. The case fatality ratio due to burns was 8.4% (77 deaths among 921 patients), with a rate of 6.4% for men and 12.2% for women. The case fatality ratio was higher among women than men regardless of the city of residence. The case fatality ratio was 3.2 and 4.4 times greater for men and women burn victims from other towns than for burn victims from Ribeirao Preto, indicating the need for additional equipment and training of medical and paramedical personnel in the initial measures to be taken with burn patients. PMID- 11900931 TI - Dirofilaria repens infection in a dog: diagnosis and treatment with melarsomine and doramectin. AB - Therapy of canine dirofilariois due to Dirofilaria repens is indicated for dogs suffering from clinical signs of this disease, such as dermal swelling, sub cutaneous nodules and pruritus. It is also important in order to decrease the risk of infection to other dogs and humans in the vicinity of the infected animal when suitable mosquito vectors are present. Combined therapy with the arsenic adulticide melarsomine and the avermectin microfilaricidal doramectin was effective in clearing infection with D. repens in a dog. The number of microfilariae dropped from 17 microl(-1) blood pre-treatment to 7 microl(-1) following the first adulticide injection and reached 0 a day after the microfilaricidal administration. The dog remained negative for D. repens microfilaremia during a follow-up period of 90 days. Euthanasia and necropsy performed 3 months after the initiation of therapy due to a progressive neoplastic disease revealed no evidence of filariae. PMID- 11900933 TI - Deliberate self-burning in Mazandaran, Iran. AB - The authors in a prospective descriptive study, via a demographic questionnaire, semi-structured interview and/or psychological autopsy examined 318 cases of self burning in Mazandaran, Iran during 3 years. The average age was 27 years and 83% of them were female. Most of them were married, home makers and with high school education. Sixty-two percent had an impulsive suicidal intention. The major motive was marital conflict. Ninety-five percent had a psychiatric diagnosis mostly adjustment disorder and 30% had a chronic physical illness. Mortality rate was 79%. High prevalence of self-burning in the young population, the pattern of demographic factors, their motivations and high prevalence of adjustment disorders highlights the need for making preventive measures, which should be focused on family structure, particularly in relation to marriage. PMID- 11900934 TI - The epidemiology of burns and smoke inhalation in secondary care: a population based study covering Lancashire and South Cumbria. AB - The epidemiology of burns and smoke inhalation in secondary care, for the population (1.6 million) of the four U.K. health authorities of Lancashire and South Cumbria is presented. Using health authority data from 1997 to 1999, it was found that 925 patients were admitted to hospital with either a primary diagnosis of burns or a primary diagnosis of smoke inhalation, in which 66% were male and 34% were female. The overall rate of admission was 0.29 per thousand. Highest rates were observed in children under the age of 5 and the elderly over the age of 75. Regression analysis confirmed an increase in admissions with increasing social deprivation.Mortality rates were shown to be highest in the over 75s. Rates of admitted burns in this study are higher than those reported from southern England. Paediatric and elderly injuries have been highlighted as high incidence groups. The epidemiology described here should assist in formulating strategies for prevention and the planning of further research. PMID- 11900935 TI - A review of the specialties that care for inpatient burns and smoke inhalation in the English counties of Lancashire and South Cumbria. AB - Prevention is by far the best strategy to minimise the burden of burns and smoke inhalation injuries on public health. However, it is inevitable that some injuries will occur despite the best attempts to prevent them. We must, therefore, optimise treatment in order to restore individuals to the best possible condition. Previous experience has shown that a wide range of specialties, many of which are untrained in burn care medicine, are involved in the care of inpatient burns/smoke inhalation victims in the UK. In light of this, a local review of which specialties care for such injuries was conducted for the population of Lancashire and South Cumbria in the north-west of England. Using population-based health authority data from 1997 to 1999, all Hospital Episodes relating to a primary diagnosis of burns or smoke inhalation were ascertained. The results showed that 41% of all burns episodes were treated by specialties other than burns/plastics. The short lengths of stay in non-plastics/burns specialties suggest that relatively minor injuries are being admitted to these units. Analysis of smoke inhalation injuries showed admission to various different specialties. Admission to burn services ensures that key specialties are available for the care of complex burn injuries. These multidisciplinary teams include burn nurses, burn surgeons and burn anaesthetists/intensivists. From the data available, it was not possible to assess the appropriateness of admission of burns and smoke inhalation injuries to the various branches of medicine. In order to assess appropriateness, we need information on severity of injury and outcome of treatment in each specialty. Further research in this area is required since it is concerning that many burns/smoke inhalation injuries are being treated by specialties with no formal training in burn care medicine. This may have major implications for service planning alongside changes in referral patterns. PMID- 11900936 TI - The UK pre-hospital management of burn patients: current practice and the need for a standard approach. AB - INTRODUCTION: In any system of burn care, first-aid, packaging and transportation of the burn injured patient from outside of hospital is a most important contribution to the successful management and outcome. This study aimed to assess the current initial care of burn patients given by the statutory ambulance services and then compare this to a survey of opinions among the plastic surgery and burns consultants in the United Kingdom (UK). METHODS: In 1999, each of the UK ambulance services was contacted via a postal questionnaire. A similar survey was sent to all of the plastic surgery consultants within the UK (taken from the specialist register) therefore, canvassing the plastic surgeons who deal less commonly with burn patients as well as the burns units. RESULTS: A total of 58% of ambulance services said that they had no treatment policy for burns patients; 97% sent patients to their nearest A&E department; 84% of services employed cooling; 12 different types of dressing were used for burn patients; 74% of services used nalbuphine hydrochloride and 97% used entonox; 74% services gave oxygen to all burn patients; 90% cannulated patients, with or without fluid administration. Plastic surgical opinion indicated that the most important aspects of basic first-aid should include: stopping the burning process; cooling (15 min (median)); airway, breathing and circulation assessment; clothing removal and dressings (clingfilm). Oxygen need not be given to all patients, but they should be kept warm and administered entonox and/or intravenous morphine. Most surgeons felt that patients should be taken to the nearest A&E and the majority of surgeons caring for this large group of patients did not have good and regular liaison with their local ambulance service. CONCLUSIONS: There seems to be a wide variation in the basic approach to the first-aid and pre-hospital care of burns patients. A significant improvement in management for this large and important group of patients is achievable, if a standard approach across all ambulance services could be achieved. PMID- 11900937 TI - Calcium induced the damage of myocardial mitochondrial respiratory function in the early stage after severe burns. AB - OBJECTIVE: To explore the role of Ca(2+) in the damage to myocardial mitochondrial respiratory function in the early stage after severe burns. METHODS: An experimental model of 30%TBSA full-thickness skin scalding was reproduced in rats. Myocardial mitochondria were isolated from control and burned rats in the 1st, 3rd, 6th, 12th and 24th hour post-burn. The mitochondrial respiratory function, contents of mitochondrial calcium ([Ca(2+)](m)), activities of mtPLA(2), mtNOS, F(0)F(1)-ATPase and cytochrome c oxidase were determined. RESULTS: (1) At the 1st hour post-burn, [Ca(2+)](m) was increased significantly and the myocardial mitochondrial respiratory function was significantly reinforced. At the same time, mitochondrial respiratory control rate (RCR) was elevated and positively correlated with [Ca(2+)](m) (r=0.8415, P<0.01). At the 3rd, 6th, 12th and 24th hour post-burn, [Ca(2+)](m) increased further to a higher level, however, the mitochondrial respiratory function was decreased from the peak value at 6h, and RCR was negatively correlated with [Ca(2+)](m). (2) The activities of mtNOS and mtPLA(2) were higher significantly at the 3rd, 6th, 12th and 24th hour post-burn than that of the control. After severe burns, mtNOS and mtPLA(2) activities were both positively correlated with [Ca(2+)](m) (r=0.8945, P<0.05; r=0.9271, P<0.01, respectively). (3) The F(0)F(1)-ATPase synthetic activity increased at the 1st hour post-burn, but it decreased to 51.4, 44.9, 77.6 and 87.4% of that of the control at the 3rd, 6th, 12th and 24th hour post burn respectively. The F(0)F(1)-ATPase hydrolytic activity decreased at the 1st hour post-burn and increased at the 3rd, however, it decreased again at the 6th, 12th and 24th hour post-burn. The activity of cytochrome c oxidase at the 3rd, 6th, 12th and 24th hour was low compared to the control. CONCLUSIONS: The changes of [Ca(2+)](m) were involved in damage to or regulation of mitochondrial respiratory function after severe burns. Appropriate increase of [Ca(2+)](m) reinforced the mitochondrial respiration at 1st hour after of burn injury, but Ca(2+) severe overload impairing F(0)F(1)-ATPase and cytochrome c oxidase directly, or, indirectly by activation of mtPLA(2) and mtNOS, might play an important role in damage to myocardial mitochondrial respiratory function at later stages after severe burns. PMID- 11900938 TI - Serum cholinesterase activity reflects morbidity in burned patients. AB - Blood samples of 200 patients with thermal injuries were drawn apparently to determine the trend of the cholinesterase activity. In relation to the severity of the injury, a characteristic decrease was noted during the first days after admission. A recovery to normal values was achieved in all survivors (150 patients) after a proportionate period of time, but in the group of non-survivors (50 patients), no complete recovery to normal levels was found. Furthermore, a significant relationship between serum cholinesterase activity and the severity of morbidity was detected, the fall of the cholinesterase activity at the very beginning was significantly higher (P<0.004) in patients who died (1.3kU/l) than in patients who survived (0.7kU/l). Already 24h after admission, the mean activity was significantly lower (P<0.003) in non-survivors (2.5kU/l) than in survivors (3.2kU/l). It seems that the serum cholinesterase is a sensitive indicator for the morbidity of patients with severe burn injuries. PMID- 11900939 TI - Long-term results of a clinical trial on dermal substitution. A light microscopy and Fourier analysis based evaluation. AB - Although dermal substitution is a main topic of current wound healing research, there is a paucity of clinical trials with a long-term clinical and histopathological evaluation. A clinical trial was conducted to perform an intra individual comparison of conventional treatment (split-thickness autograft) to a collagen/elastin dermal substitute in combination with an autograft. Promising results with this substitute were obtained with respect to dermal organisation and scar elasticity in animal studies and clinical trials with a short-term follow-up. Twenty-nine of the 42 pairs of the burn wounds and 28 of the 44 pairs of the scar reconstructions enrolled in the study were biopsied after 1 year. Promising but not statistically significant differences were found between substituted groups and control groups for epidermal thickness, basement membrane maturation, rete ridges (P=0.055), fibroblasts, myofibroblasts, inflammatory cells, vessels and extracellular matrix maturation. An objective and accurate technique, Fourier analysis, was used to evaluate collagen bundle orientation and packing. However, no statistically significant differences were obtained for these parameters. This microscopic evaluation provided no convincing evidence for a long-term effectiveness of a dermal substitute despite promising data over a short-term in in vitro and in vivo studies with the same material. PMID- 11900940 TI - Serum lactate, not base deficit, rapidly predicts survival after major burns. AB - BACKGROUND: Clinical studies document correlation of serum lactate and base deficit with mortality in trauma and sepsis. No study of the prognostic value of these two serum markers has been reported in burn injury. METHODS: Resuscitation data from 49 patients admitted to the adult Burn ICU were analyzed. Lactate and base deficit were analyzed upon admission and every 2h during the initial 48 h after admission. Resuscitation was managed per standard routine, blinded to these data, guided by the Parkland formula. Initial statistical analysis with Cox's regression model was used to determine the relationship between survival, resuscitation parameters, and demographics. Then, a logistic regression was used to determine if any of these variables were quickly predictive (initial values) of the risk of death. RESULTS: Two variables were predictive of mortality by the Cox regression model: (1) serum lactate value and (2) patient age. Furthermore, analysis by logistic regression revealed that the initial serum lactate value was separately predictive of mortality. CONCLUSION: In this study, serum lactate but not base deficit, was a predictor of mortality following major burns. Moreover, initial serum lactate values were also predictive of mortality separately. PMID- 11900941 TI - The importance of initial management: a case series of childhood burns in Vietnam. AB - The success of treatment of childhood burns is critically dependent on how well the initial management is performed. In this case series of 695 children with burns transferred to the National Burn Institute (NBI) in Hanoi from peripheral hospitals, the initial management of each patient was assessed for the following initial management measures: removal of the cause and immediate cooling with water at the accident site; and pain relief, dry dressing, administration of oxygen, and adequate fluid replacement at the peripheral hospital. Overall, 61 of the 695 children died, but of the 95 patients who received all of these initial management measures, all survived. There were no cases of irreversible shock, acute renal failure, or multiple organ failure in the patients who received adequate initial management. Provision of adequate initial management was also significantly protective against septicaemia. Thus in this group of subjects who survived until admission, effective initial management significantly reduced the risk of death and other complications such as irreversible shock, septicaemia and multiple organ failure. PMID- 11900942 TI - The importance of immediate cooling--a case series of childhood burns in Vietnam. AB - Numerous experimental studies have shown several benefits of treating burns by the immediate application of cool water. In this study of 695 children with burns, treated in the National Burn Institute (NBI), Hanoi, Vietnam, patients were assessed on admission according to first aid measures at the time of injury, i.e. the removal of the cause and immediate cooling with cold water. A total of 33% of the children who had had immediate cooling of the burn with water had deep burns, compared with 49% of the children who had not had immediate cooling. The prevalence ratio of deep burns was thus 0.68 (95% confidence interval (CI) 0.55 0.85); that is, there was an estimated reduction of 32% in the need for skin grafting, a reduction which was statistically significant. After adjusting for the effect of cooling the burn, removal of the causal agent reduced the odds of requiring skin grafting, but the reduction was not statistically significant. It is concluded that early cooling will prevent a significant percentage of superficial burns from progressing to deep burns. This will not only reduce the probability that skin grafting and expensive treatment will be required, but will reduce the risk of other consequences of deep burns, which may be fatal. Public health programs to promote immediate cooling of burns with cool water are at least as important as subsequent medical and surgical treatment in determining the outcome of burns in children. PMID- 11900943 TI - Treatment modalities for post-burn axillary contractures and the versatility of the scapular flap. AB - Inappropriate treatment of axillary burns frequently results in adduction contractures. In this clinical study we have reviewed 32 patients with different types of axillary post-burn adduction contractures. We have used a variety of surgical treatments for reconstruction of axillary contracture releasing defects such as simple grafting, Z-plasties and locally pedicled flaps. Among these alternatives, we preferred to use scapular island flap most frequently. In addition to conventional harvest of this flap, extension of its pedicle up to the subscapular ramification by passing it through the triangular space allowed its transfer even to the anterior axillary line defects in a vertical orientation without pedicle kinking. In conclusion, the island scapular flap is a good choice for reconstruction of all types of axillary contracture, releasing defects with satisfactory results in terms of function and cosmesis. PMID- 11900944 TI - Toxic shock syndrome following cessation of prophylactic antibiotics in a child with a 2% scald. AB - Toxic shock syndrome (TSS) is a rare but serious complication of burns in children. Symptoms usually start within 3 days of the burn injury and even children with minor burns can be susceptible. Prompt diagnosis and rapid treatment is crucial in limiting the morbidity associated with this condition. We report here a 19-month-old child with a 2% scald who developed TSS following cessation of a 5-day course of flucloxacillin. This case highlights a number of issues regarding the use of prophylactic antibiotics and TSS as well as illustrating the continuing need to educate parents concerning the importance of seeking a prompt medical opinion if the child becomes ill following even a minor burn injury. PMID- 11900945 TI - The use of linezolid in the treatment of vancomycin-resistant enterococcal septicaemia in two patients with burn injuries. AB - Vancomycin-resistant enterococci (VRE) are multi-resistant micro-organisms that have emerged as important nosocomial pathogens during the last decade. Emergence of this organism has been blamed mainly on the increased and inappropriate use of antibiotics, in particular, the cephalosporins and the glycopeptide, vancomycin. Burns patients are highly vulnerable to acquiring VRE infections, being both debilitated and immunocompromised, and often receiving antibiotics that further diminish their intrinsic microbial flora. We report on two patients with large burn injuries who acquired vancomycin-resistant enterococcal septicaemia during their in-patient stay. Both patients were successfully treated using the antibiotic, linezolid. Linezolid is the first in a new class of antibiotics known as the oxazolidinones whose mode of action inhibits early bacterial protein synthesis. Linezolid has a spectrum of activity against Gram-positive micro organisms including methicillin-resistant Staphylococcus aureus (MRSA) and VRE, and can provide a useful treatment alternative to the glycopeptides. PMID- 11900946 TI - Colonic necrosis and perforation following oral sodium polystyrene sulfonate (Resonium A/Kayexalate in a burn patient. PMID- 11900948 TI - Fillet flap from the ring finger dorsum for salvaging the little finger in electrically burned hand. PMID- 11900947 TI - The use of a PEG tube in a burn centre. AB - Patients with extensive burn injuries frequently require supplemental tube feeds in order to maintain caloric balance. Conventionally, nutrition is supplied via a naso-gastric tube (NGT). However, NGTs cause discomfort and numerous complications have been described, particularly following prolonged use. An alternative route to the gastro-intestinal tract comprises the percutaneous endoscopic gastrostomy (PEG) tube. This study describes by means of a retrospective analysis our experiences with the PEG tube in comparison with the NGT in a burn centre. Twelve burn patients, including two children and two patients with toxic epidermal necrolysis (TEN), were treated with a PEG tube. We could find no contra-indications to the placement of PEG tubes in this group. Placement of the tube through partial or full-thickness burn wounds did not give rise to complications. In a comparable group of 12 consecutive patients who were fed using a NGT, the NGT did give substantial discomfort to the patients and caused complications, especially in the patients with TEN. The complications that occurred during the use of a PEG tube were mainly caused by the small diameter, single-lumen design. Our experience suggests that the PEG tube is preferable to the NGT for patients who require prolonged feeding. For burn patients, modification of the design of the tube to include two exchangeable lumens of sufficient diameter, would improve performance. PMID- 11900949 TI - Unusual type of burn injury caused by industrial bakery ovens. PMID- 11900950 TI - Current attitudes to burns resuscitation in the UK. PMID- 11900951 TI - Dermal cellular inflammation in burns: an insight into the function of dermal microvascular anatomy. PMID- 11900953 TI - Value of existing serological tests for identifying badgers that shed Mycobacterium bovis. AB - In the UK there has been a sharp rise in the incidence of bovine tuberculosis since the early 1990s and the badger has been identified as an important wildlife reservoir for this infection. Infected badgers can excrete Mycobacterium bovis, putting other badgers and cattle at risk of becoming infected. Vaccination has been proposed as an approach to reducing the excretion of M. bovis by tuberculous badgers. In order to evaluate the efficacy of a badger vaccine it will be necessary to accurately determine the number of badgers excreting M. bovis without removing them for post-mortem evaluation. The existing live tests for tuberculosis in the badger (culture, indirect ELISA, Western blot) have not been assessed for their ability to detect badgers excreting M. bovis. Over the past 18 years, badgers from 31 social groups have been trapped and sampled in a study area of the Cotswold escarpment. We have examined the serological responses of 128 badgers trapped between 1985 and 1998 from social groups where M. bovis infection was endemic. These responses were compared with culture from faeces, urine, tracheal aspirates and bite wound swabs taken from these animals while alive. ELISA was found to be more sensitive than Western blot in detecting badgers excreting M. bovis. The majority of culture-positive badgers excreted M. bovis intermittently over the period of study. As a result, there was only a 27.5% chance of sampling a badger for culture when it was excreting M. bovis. In contrast, a positive ELISA result correctly predicted 68.2% of badgers with a history of excreting M. bovis. In the absence of alternative live tests for the badger, the Brock Test indirect ELISA appears to be more valuable than culture for measuring the effect of vaccination on reducing the number of badgers at risk of transmitting tuberculosis. PMID- 11900955 TI - Sero types, phage types and antibiotic susceptibilities of Salmonella strains isolated from horses in The Netherlands from 1993 to 2000. AB - We studied 232 Salmonella strains from horses with salmonellosis in The Netherlands, isolated in the period from 1993 to 2000 in order to provide insight in the dynamics of sero-, phage types (pt) and antibiotic susceptibilities over time. The strains were tested for susceptibility to seven antimicrobial agents using the agar diffusion method. In addition, the isolates were sero typed and Salmonella enterica subspecies enterica Typhimurium and Enteritidis strains were further phage typed. S. Typhimurium strains of phage type 506 and 401 (both classified as DT 104 in the English phage typing system) were additionally tested for their susceptibility to chloramphenicol (C), streptomycin (S) and sulfonamides (Su). Resistance was common against tetracycline and ampicillin. Most strains were susceptible to enrofloxacin (Enr) and ceftiofur (Cef). Resistance to tetracycline (T), kanamycin (K), ampicillin (A) and trimethoprim/sulfonamide (Sxt) combinations decreased from 1993 to 2000, whereas the resistance to gentamicin (G), ceftiofur and enrofloxacin was stable over time. S. Typhimurium was the predominant serovar and showed more (multiple) resistance compared to other Salmonella serovars. Sixteen different resistance patterns were found, with resistance to T alone and the combination of ACSSuT and AKSxtT being the most common. The multiresistant S. typhimurium phage type 506 (DT 104) was the most common phage type isolated from horses and most of these strains showed the pentadrug resistance pattern ACSSuT. The S. Typhimurium phage type 401 (DT 104) was also found frequently with an ASSuT resistance pattern. The most common S. Typhimurium phage types in horses corresponded with those found in humans, pigs and cattle in the same period in The Netherlands. PMID- 11900954 TI - A role for the Clostridium perfringens beta2 toxin in bovine enterotoxaemia? AB - Non-enterotoxigenic type A Clostridium perfringens are associated with bovine enterotoxaemia, but the alpha toxin is not regarded as responsible for the production of typical lesions of necrotic and haemorrhagic enteritis. The purpose of this study was to investigate the putative role of the more recently described beta2 toxin. Seven hundred and fourteen non-enterotoxigenic type A C. perfringens isolated from 133 calves with lesions of enterotoxaemia and high clostridial cell counts (study population) and 386 isolated from a control population of 87 calves were tested by a colony hybridisation assay for the beta2 toxin. Two hundred and eighteen (31%) C. perfringens isolated from 83 calves (62%) of the study population and 113 (29%) C. perfringens isolated from 51 calves (59%) of the control population tested positive with the beta2 probe. Pure and mixed cultures of four C. perfringens (one alpha+beta2+, one alpha+enterotoxin+ and two alpha+) were tested in the ligated loop assay in one calf. Macroscopic haemorrhages of the intestinal wall, necrosis and haemorrhages of the intestinal content, and microscopic lesions of necrosis and polymorphonuclear and mononuclear cell infiltration of the intestinal villi were more pronounced in loops inoculated with the alpha and beta2-toxigenic C. perfringens isolate. These results suggest in vivo synergistic role of the alpha and beta2 toxins in the production of necrotic and haemorrhagic lesions of the small intestine in cases of bovine enterotoxaemia. However, isolation of beta2-toxigenic C. perfringens does not confirm the clinical diagnosis of bovine enterotoxaemia and a clostridial cell counts must still be performed. PMID- 11900956 TI - Characterization of the carrier state in porcine reproductive and respiratory syndrome virus infection. AB - Porcine reproductive and respiratory syndrome (PRRS) virus infection results in clinically normal, but persistently infected animals. An understanding of the carrier state is necessary for prevention, control and/or elimination of PRRS virus. The objective of this experiment was to estimate the proportion of PRRS virus carriers over time and determine which combination of sample and diagnostic assay could most effectively identify persistently infected animals. Sixty 3-week old pigs were inoculated with PRRS virus ATCC VR-2332 and followed for up to 105 days post-inoculation (PI). Sixty age-matched animals served as uninoculated controls. Samples (serum, peripheral blood leukocytes, oropharyngeal scrapings, tonsil, bronchoalveolar lavage, lung tissue and tracheobronchial lymph nodes) were collected periodically and tested for evidence of PRRS virus infection by virus isolation (VI), swine bioassay and reverse transcriptase-nested polymerase chain reaction (RT-nPCR). The PRRS virus-specific antibody response was monitored with a commercial enzyme-linked immunosorbent assay (ELISA). Overall, PRRS virus was found in 51 of the 59 (84%) necropsied animals by VI or swine bioassay between 63 and 105 days PI, including 10 of the 11 (91%) of animals at day 105 PI. RT-nPCR on oropharyngeal scrapings was the most effective combination of assay and sample for detecting carriers. There was no significant difference in the antibody response of carrier vs. non-carrier animals. Infectious PRRS virus is present in most pigs the first 105 days following infection. Antibody response, as measured by a commercial ELISA, cannot be used to determine carrier status. RT-nPCR is a useful tool for detection of carriers, but diagnostic sample selection is critical if false negative results are to be avoided. PMID- 11900957 TI - Resistance and susceptibility to Marek's disease: nitric oxide synthase/arginase activity balance. AB - The metabolic NO pathway, catalyzed by the enzyme NO synthase in macrophages, is a key defense element against viruses and tumors. However, arginase is an other enzyme able to metabolize the substrate L-arginine, and the two enzymes are alternatively regulated by Th1 and Th2 cytokines in murine macrophages. Marek's disease is characterized by strong immunosuppression and development of T-cell lymphomas in chickens. Inoculation of the very virulent strain of MDV RB-1B induced strong and long-lasting arginase macrophage-dependent activity, which was inhibited by L-norvaline in vitro, but induced low NO production in monocytes and splenocytes from highly susceptible B(13)/B(13) chickens. By contrast, in B(21)/B(21) chickens genetically resistant to tumor development, RB-1B induced a weak and transient increase in arginase activity and strong NO production. The vaccinal HVT strain did not induce any arginase activity in monocytes or splenocytes. Moreover, vaccination with HVT prevented tumor appearance after RB 1B challenge and increase in arginase activity, but favored NO production in susceptible chickens. Differential expression of NO synthase and arginase was modulated in chicken macrophages, with IFN-gamma and LPS being strong inducers of both, depending on the type of macrophage, and TGF-beta 1 and PGE(2) stimulating only arginase activity. This increase in arginase activity in macrophages from chickens inoculated with Marek's disease virus might thus be due to a direct effect of the virus on macrophages, possibly through viral products, or to indirect effects on the cytokine balance. PMID- 11900958 TI - Cloning and expression of the superoxide dismutase gene of the feline strain of Porphyromonas gingivalis: immunological recognition of the protein by cats with periodontal disease. AB - Recent evidence suggests that feline members of the genus Porphyromonas are of consequence in periodontal disease in cats. Several possible virulence factors from feline strains of Porphyromonas gingivalis have been described that have similarities to those of human P. gingivalis. Both human and feline strains of P. gingivalis produce superoxide dismutase (SOD) which has been proposed as modulator of the inflammatory response during infection. The objective of this study was to clone the superoxide dismutase gene of feline P. gingivalis, to compare the characteristics of its product with that of the native enzyme and to determine its immunoreactivity in cats with periodontal disease. The sod gene of the feline strain Veterinary Pathology and Bacteriology (VPB) 3457 of P. gingivalis was amplified by PCR and cloned in frame with the alpha-peptide of the LacZ gene of E. coli in plasmid pUC19. This construct expressed SOD activity in E. coli with characteristics similar to those of the native SOD enzyme of P. gingivalis human strain 381 and the parent feline strain VPB 3457. The recombinant SOD had an apparent molecular weight of 54,700+/-1300 (S.E.M.) and was inactivated by 5mM hydrogen peroxide but not by 2mM KCN. There was a significant association (P=0.005) between the immunoreactivity of cats to P. gingivalis VPB 3457 soluble whole cell proteins on immunoblots and their responsiveness to the SOD protein. This suggests that cats showing a marked serum responsiveness to P. gingivalis itself, react to the SOD enzyme and further supports the role of feline P. gingivalis in periodontal disease. PMID- 11900959 TI - Phylogenetic analysis and PCR detection of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum based on the flagellin gene. AB - The flagellin genes (fliC) of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene (fliC) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I (), are more closely related to Bacillus subtilis than to Clostridium difficile, which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei, C. haemolyticum, C. novyi types A and B, and C. septicum. PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia. PMID- 11900960 TI - Evaluation of the long-term immune response in cattle after vaccination against paratuberculosis in two Dutch dairy herds. AB - In the past decades, vaccination against paratuberculosis in cattle was performed in The Netherlands only on a limited scale. Because of its interference with the diagnosis of bovine tuberculosis, vaccination was restricted to herds with a high prevalence of clinical cases of paratuberculosis and was meant to aid in the economical survival of the farm. Recently, a voluntary paratuberculosis certification program has started, based in part on serological screening of cattle of at least 3 years of age. Herds that have been vaccinated against paratuberculosis are, therefore, likely to encounter problems when entering this program. The aim of this study was to evaluate the immune response resulting from vaccination with a heat-killed paratuberculosis vaccine. Over a period of 12-14 years, new-born calves were vaccinated in two herds. The B-cell response was evaluated using both the complement-fixation test (CFT) and an enzyme-linked immunosorbent assay (ELISA) and the cell-mediated immune response was evaluated using the gamma-interferon assay. Data obtained show a marked and prolonged effect of the vaccination on both cellular and humoral immune responses, in particular to the paratuberculosis antigen but also to the bovine tuberculosis antigen, using the respective tests. These responses were detected rapidly after vaccination. The individual responses were highly variable between animals with respect to both the level and to the duration of the evoked immune response. No relation between the results obtained with the ELISA and the CFT was observed. In conclusion, for a large number of vaccinated cattle, a long lasting interference is to be expected with the presently available immunodiagnostic methods for both bovine tuberculosis and paratuberculosis. PMID- 11900961 TI - Peroxisome proliferator-activated receptor gamma (PPARgamma) and its ligands: a review. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of a class of nuclear hormone receptors intimately involved in the regulation of expression of myriad genes that regulate energy metabolism, cell differentiation, apoptosis and inflammation. Although originally discovered as a pivotal regulator of adipocyte differentiation, the roles that this transcription factor play in physiology and pathophysiology continue to grow as researchers discover its influence in the function of many cell types. This review highlights the roles that PPARgamma play in the regulation of gene expression associated with normal cell physiology as well as the pathophysiology of multiple diseases including obesity, diabetes and cancer. Additionally, naturally occurring and pharmaceutical ligands for the receptor as well as the potential role of PPARgamma as the receptor responsible for fatty acid-induced effects on gene expression will be described. PMID- 11900962 TI - Effects of aging and weaning on mRNA expression of leptin and CCK receptors in the calf rumen and abomasum. AB - In order to know the effects of weaning and volatile fatty acid feeding on gastric leptin expression, we investigated the expression of leptin and CCK receptor mRNA in the bovine rumen, abomasum and duodenum using RT-PCR in 3-week old pre-weaning, 13-week-old post-weaning and adult animals. Leptin mRNA was expressed in the rumen and abomasum of 3-week-old pre-weaning animals, but it was abolished in 13-week-old and adult animals. In the duodenum, leptin expression was observed in the 3-, 13-week-old and adult animals. In the rumen, CCK(A) receptor mRNA was expressed in 3-week-old animals, but not in 13-week-old and adult animals. In the abomasum, CCK(B) receptor expression gradually decreased from 3-week-old to adult animals. Expression of CCK(B) receptor and of CCK(A) receptor was slight in the rumen and abomasum, respectively. In the next study, we examined the effect of weaning of 6 weeks or non-weaning (fed on milk replacer alone (milk) or milk replacer with volatile fatty acids (milk+VFA) until 13 weeks old) on leptin mRNA expression in the rumen and abomasum. In 13-week-old calf rumen and abomasum, leptin mRNA expression was detected in non-weaning milk-fed animals at 13 weeks old, although it was not observed in weaning and non-weaned milk+VFA-fed animals. The change in CCK(A) receptor expression in the rumen was similar to those of leptin mRNA expression. CCK(B) receptor transcription in the abomasum of milk-fed animals was higher than that of the weaning and milk+VFA-fed animals. These results indicate that leptin expression is coincident with CCK receptor expression in calf stomachs, and that leptin and CCK receptor mRNA expression are affected by the change in the physiological status brought about by weaning and VFA feeding. PMID- 11900963 TI - Effect of apoptosis on phagocytosis, respiratory burst and CD18 adhesion receptor expression of bovine neutrophils. AB - Polymorphonuclear neutrophil leukocytes (PMN) play an important role in intramammary defense against infections by Escherichia coli. During mastitis, PMN are confronted with various inflammatory mediators that can modulate their function. In severely diseased cows, increased concentrations of lipopolysaccharide (LPS) and tumor necrosis factor (TNF)-alpha (TNF-alpha) are detected in plasma. Binding of LPS to membrane bound CD14 molecules on monocytes cause release of inflammatory mediators such as TNF-alpha. Because apoptosis of PMN promotes resolution of inflammation and because the LPS and TNF-alpha response in milk and blood is related to the severity of E. coli mastitis, the effect on apoptosis of bovine PMN of increased concentrations LPS and TNF-alpha was studied together with the functionality of apoptotic PMN. Bovine PMN apoptosis, as determined with annexin-V, was induced with high concentrations of either LPS (1000 and 10,000ng/mL) or TNF-alpha (10,000ng/mL) in whole blood following a 6h incubation at 37 degrees C. The apoptosis inducing effect of LPS on PMN was not inhibited following coculture with either anti-bovine TNF-alpha or anti-ovine CD14 monoclonal antibodies. When compared to controls, apoptotic PMN had a similar level of CD18 expression but lacked phagocytic and respiratory burst activity. This is the first study reporting the effects of apoptosis on bovine PMN function. These functional impairments in apoptotic PMN could be important in contributing to the establishment of intramammary infection. Well functioning PMN could finally determine the severity of mastitis following an invasion of bacteria in the mammary gland. PMID- 11900964 TI - Expression and localization of IGF family members in bovine antral follicles during final growth and in luteal tissue during different stages of estrous cycle and pregnancy. AB - The objectives of the study were to monitor the detailed pattern for mRNA expression (RT-PCR and RPA) of IGFs, IGFR-1, IGFBPs, GHR and localization of protein (immunohistochemistry) for IGF-1 and IGFR-1 in bovine follicle classes during final maturation and different corpus luteum (CL) stages during estrous cycle and during pregnancy. A relative high expression of IGF-1 in theca interna (TI) was observed before selection (E<0.5ng/mL). In GC, mRNA expression increased after selection. In contrast, IGF-2 was mainly expressed in the TI. The IGFR-1 mRNA was present in the TI and GC with increasing levels during final development. The expression results were confirmed by localization of IGF-1 and IGFR-1 proteins in GC and TI. There is clear evidence for the local expression of IGFBPs in TI and GC compartment with clear regulatory differences. In CL, the highest mRNA expression of IGF-1, IGF-2 and IGFR-1 was observed during early luteal phase, followed by a decrease, and then by a tendency of an increase during the mid and late luteal phases of the cyclic CL. This level remained low during pregnancy. Intense immunostaining for IGFR-1 in CL was observed mainly in large luteal cells. Evidence for a mRNA for all six IGFBPs were obtained with distinct differences for BP-3, -4 and -5. In conclusion, this comprehensive study gives clear evidence for an important role of the IGFs and IGFBPs in bovine follicular development and CL function. The relative amounts of IGFBPs may ultimately determine ovarian IGF action. PMID- 11900965 TI - Effect of short-term feed restriction and refeeding on serum concentrations of leptin, luteinizing hormone and insulin in ovariectomized gilts. AB - Ovariectomized gilts were either placed on full feed (FF) or restricted to one third of the full feed amount (RST) for 7 days. Blood samples were taken through jugular catheters every 15 min for 4 h at the end of the 7-day period. Then dietary treatments were reversed and 7 days later samples were taken as before. Serum concentrations of leptin, insulin and luteinizing hormone (LH) were determined by radioimmunoassay. LH pulse frequency and mean serum leptin and insulin concentrations were lower (P < 0.01) in RST than FF gilts. Reversal of treatment reversed the patterns of hormone secretion. These results confirm previous observations that feed restriction can inhibit pulsatile LH secretion and also decrease leptin and insulin secretion. PMID- 11900966 TI - Seasonal differences in progesterone production by luteinized bovine thecal and granulosa cells. AB - This study examined seasonal differences in progesterone (P4) production by granulosa cells (GC) and thecal cells (TC) that were luteinized in vitro during the winter or the summer; it also compared plasma P4 concentrations of lactating dairy cows in the two seasons. First-wave dominant follicles obtained from Holstein cows were dissected on day 6 of the cycle, GC and TC were separated, enzymatically dispersed, and cultured for 9 days in media containing 1% fetal calf serum, forskolin (10 micromol/mL) and insulin (2 microg/mL), to induce cell luteinization. All experimental procedures were identical and characteristics of the follicles were similar in the two seasons. During 9 days of culture, P4 production by luteinized GC was higher in winter than in summer, but the difference only tended to be significant. In contrast, luteinized TC produced three times as much P4 in winter as in summer (324 versus 100 ng/10(5)cells). In the in vivo experiment, P4 concentrations in plasma collected during entire estrous cycles in winter and summer were compared. The cows were, on average, at 70 days postpartum and yielded similar amounts of milk. Concentrations of progesterone in plasma were significantly higher in winter than in summer; during the mid-luteal phase the difference between the two seasons was 1.5 ng/mL. These results indicate that chronic effects of heat-stress are possibly carried over from an impaired follicle to an impaired corpus luteum (CL), and that luteinized TC are more susceptible to heat-stress than luteinized GC. PMID- 11900967 TI - Real-time RT-PCR quantification of insulin-like growth factor (IGF)-1, IGF-1 receptor, IGF-2, IGF-2 receptor, insulin receptor, growth hormone receptor, IGF binding proteins 1, 2 and 3 in the bovine species. AB - Reverse transcription (RT) followed by polymerase chain reaction (PCR) is the technique of choice for analysing mRNA in extremely low abundance. Real-time RT PCR using SYBR Green I detection combines the ease and necessary exactness to be able to produce reliable as well as rapid results. To obtain highly accurate and reliable results in a real-time RT-PCR a highly defined calibration curve is needed. We designed and developed nine different calibration curves, based on recombinant DNA plasmid standards and established them on a constant real-time PCR platform for the following factors: growth hormone receptor (GHR), insulin like growth factor (IGF)-1, IGF-1 receptor (IGF-1R), IGF-2, IGF-2 receptor (IGF 2R), insulin receptor (INSR), and IGF-binding proteins (IGF-BP) 1, 2 and 3. Developed assays were applied in the LightCycler system on bovine ileum and liver total RNA and showed high specificity and sensitivity of quantification. All assays had a detection limit of under 35 recombinant DNA molecules present in the capillary. The SYBR Green I determination resulted in a reliable and accurate quantification with high test linearity (Pearson correlation coefficient r > 0.99) over seven orders of magnitude from <10(2) to >10(8) recombinant DNA start molecules and an assay variation of maximal 5.3%. Applicability of the method was shown by analysing mRNA levels in newborn calves: mRNA concentrations per gram tissue of mRNAs of IGF-1, IGF-1R, IGF-2, IGF-2R, GHR, INSR, and IGF-BP1, 2 and 3 were all different between in liver and ileum and the traits all exhibited individual differences. PMID- 11900968 TI - Effect of intravenous infusion of recombinant ovine leptin on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine in growing prepubertal ewe lambs. AB - In sheep, serum concentrations of leptin change congruently with increases or decreases in nutritional status, while intracerebroventricular infusions of leptin dramatically suppress feed intake in well-fed lambs, and may also increase growth hormone (GH), and/or luteinizing hormone (LH) in undernourished lambs. The objective of the present study was to determine the effects of peripherally delivered ovine leptin, via intravenous infusions, on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine. Twelve ewe lambs weighing 29.4 +/- 0.7 kg were infused intravenously with a linearly increasing dose of leptin or saline (n = 6 per group) for 10 days, reaching a maximum dose delivered of 0.5mg/h on day 10. Feed intake was assessed twice daily, and blood samples were collected every 10 min for 6 h on days 0, 2, 5, 8, and 10. Serum concentrations of leptin increased in leptin-treated lambs by day 2 (P = 0.05), and continued to increase to concentrations 9-fold greater than saline-infused lambs by day 10 (P < 0.001). Despite the substantial increase in serum leptin, feed intake did not differ between leptin and saline-infused lambs except on day 3.5 (P = 0.01). Furthermore, intravenous infusions of leptin did not significantly influence serum concentrations of insulin, cortisol, IGF-1, thyroxine, LH, or GH. Collectively, these observations contrast with the potent hypophagic effects of leptin when delivered intracerebroventricularly into well fed lambs. The reasons for the disparate response of lambs treated intravenously with leptin, versus that reported for lambs treated intracerebroventricularly with leptin are not known, but may provide insight into the mechanism(s) of leptin resistance. PMID- 11900969 TI - Exercise-induced hyperkalemia in hypothyroid dogs. AB - We investigated the effect of hypothyroidism in dogs on (1) the Na+-, K+ -ATPase concentration in skeletal muscle, and (2) potassium (K+) homeostasis at rest and during exercise. Prior to and 1 year after induction of hypothyroidism by surgery and subsequent radiothyroidectomy, the Na+-, K+ -ATPase concentrations were quantified in biopsies of sternothyroid muscles of seven Beagle dogs by measuring [3H]ouabain binding capacity. In addition, plasma K+ concentrations were measured at rest and after treadmill exercise in six hypothyroid and seven euthyroid Beagle dogs. During hypothyroidism, the mean Na+ -, K+ -ATPase concentration in muscle biopsies was 41% lower than during euthyroidism. The mean resting plasma K+ value of the hypothyroid dogs was significantly (14%) higher than that of the euthyroid dogs. In the hypothyroid dogs, plasma K+ concentration increased significantly during exercise, whereas there was no rise in the euthyroid dogs. The rise in plasma K+ concentration could not be ascribed to muscle damage, as plasma creatine kinase concentrations remained within reference range. Also renal K+ retention was an unlikely explanation, as plasma aldosterone concentration and plasma renin activity rather increased than decreased during exercise. In conclusion, hypothyroid dogs tend to develop hyperkalemia during exercise, which for a large part can be explained by the severe reduction of the Na+ -, K+ ATPase capacity in the skeletal muscle pool. PMID- 11900970 TI - Xenopus marginal coil (Xmc), a novel FGF inducible cytosolic coiled-coil protein regulating gastrulation movements. AB - Gastrulation in vertebrates is a highly dynamic process driven by convergent extension movements of internal mesodermal cells, under the regulatory activity of the Spemann-Mangold or gastrula organizer. In a large-scale screen for genes expressed in the organizer, we have isolated a novel gene, termed Xmc, an acronym for Xenopus marginal coil. Xmc encodes a protein containing two widely spaced evolutionarily non-conserved coiled coils. Xmc protein is found in vesicular aggregates in the cytoplasm and associated with the inner plasma membrane. We show that Xmc is expressed in a dynamic fashion around the blastoporal circumference, in mesodermal cells undergoing morphogenetic movements, in a pattern similar to FGF target genes. Likewise, Xmc expression can be induced by ectopic XeFGF signaling and the early mesodermal expression is dependent on FGF receptor-mediated signaling. Morpholino-mediated translational 'knock-down' of Xmc results in embryos that display a reduced elongation of the antero-posterior axis and in a pronounced inhibition of morphogenetic movements in embryos and dorsal marginal zone explants. Xmc loss-of-function does not interfere with mesoderm induction or maintenance per se. Our results suggest that Xmc is a novel FGF target gene that is required for morphogenetic movements during gastrulation in Xenopus. PMID- 11900971 TI - Rotatin is a novel gene required for axial rotation and left-right specification in mouse embryos. AB - The genetic cascade that governs left-right (L-R) specification is starting to be elucidated. In the mouse, the lateral asymmetry of the body axis is revealed first by the asymmetric expression of nodal, lefty2 and pitx2 in the left lateral plate mesoderm of the neurulating embryo. Here we describe a novel gene, rotatin, essential for the correct expression of the key L-R specification genes nodal, lefty and Pitx2. Embryos deficient in rotatin show also randomized heart looping and delayed neural tube closure, and fail to undergo the critical morphogenetic step of axial rotation. The amino acid sequence deduced from the cDNA is predicted to contain at least three transmembrane domains. Our results show a novel key player in the genetic cascade that determines L-R specification, and suggest a causal link between this process and axial rotation. PMID- 11900972 TI - Homeostatic regulation of germinal stem cell proliferation by the GDNF/FSH pathway. AB - Stem cell regulatory mechanisms are difficult to study because self-renewal and production of differentiated progeny, which are both strictly controlled, occur simultaneously in these cells. To focus on the self-renewal mechanism alone, we investigated the behavior of germinal stem cells (GSCs) in progeny-deficient testes with defective GSC differentiation. In these testes, we found that the proliferation of undifferentiated spermatogonia, some of which are GSCs, was accelerated by high concentrations of glial cell line-derived neurotrophic factor (GDNF). Furthermore, we found that follicle-stimulating hormone (FSH) stimulation via homeostatic control was one of the major regulators of GDNF concentration. These results suggest that in mammalian testes, GSC proliferation and population size are regulated homeostatically by the GDNF/FSH pathway. PMID- 11900973 TI - Identification of novel Drosophila neural precursor genes using a differential embryonic head cDNA screen. AB - During Drosophila neuroblast lineage development, temporally ordered transitions in neuroblast gene expression have been shown to accompany the changing repertoire of functionally diverse cells generated by neuroblasts. To broaden our understanding of the biological significance of these ordered transitions in neuroblast gene expression and the events that regulate them, additional genes have been sought that participate in the timing and execution of these temporally controlled events. To identify dynamically expressed neural precursor genes, we have performed a differential cDNA hybridization screen on a stage specific embryonic head cDNA library, followed by whole-mount embryo in situ hybridizations. Described here are the embryonic expression profiles of 57 developmentally regulated neural precursor genes. Information about 2389 additional genes identified in this screen, including 1614 uncharacterized genes, is available on-line at 'BrainGenes: a search for Drosophila neural precursor genes' (http://sdb.bio.purdue.edu/fly/brain/ahome.htm). PMID- 11900974 TI - Cloning and developmental expression of a novel, secreted frizzled-related protein from the sea urchin, Strongylocentrotus purpuratus. AB - Wnt proteins and their receptors, members of the frizzled protein family, play a key role in regulating a wide range of developmental processes. Recently, putative regulators of Wnt signaling known as secreted frizzled-related proteins (SFRPs) have been identified in several vertebrates. Here, we describe the cloning of a novel SFRP (suSFRP1) from the sea urchin, Strongylocentrotus purpuratus. SuSFRP1 contains a putative signal sequence, four cysteine-rich domains and a single Ig domain. The developmental expression of suSFRP1 mRNA is highly dynamic and can be separated into three phases: (1) abrupt accumulation in most or all cells of the embryo at the early blastula stage; (2) restriction of expression to the prospective endoderm and animal pole region of the gastrula; and (3) expression in prospective muscle cells of the coelomic pouches during late embryogenesis. PMID- 11900975 TI - Expression of stef, an activator of Rac1, correlates with the stages of neuronal morphological development in the mouse brain. AB - STEF (Sif- and Tiam1-like exchange factor), a guanine nucleotide exchange factor, was identified as a candidate molecule in regulation of neural development. The STEF gene product specifically activates Rac1, a member of the Rho-like small G proteins. Here we report the detailed examination of the expression profile of the stef gene in the mouse brain. In situ hybridization revealed that the stef gene was expressed in a stage- and region-specific manner in the mouse brain; it was expressed during certain developmental stages in the cerebral cortex, the olfactory bulb, the rostral migratory pathway (RMP) and the hippocampus. In the cerebral cortex, stef transcripts were detected in migrating cells in the intermediate zone as well as neurons in the cortical plate. While the expression in the cerebral cortex was reduced at adult stages, considerable expression was found to be maintained in other regions (RMP, olfactory bulb, hippocampal formation), which are the tissues where neurons continue to undergo morphological remodeling including cellular migration, neurite extension and synapse formation even in adults. Thus, stef gene expression appears to correspond to neuronal morphological changes. PMID- 11900976 TI - Cloning and expression pattern of the lysozyme C gene in zebrafish. AB - Here, we report isolation and developmental expression pattern of the zebrafish lysozyme C gene. Amino acid sequence analysis showed that the zebrafish lysozyme C protein shared approximately 37-80% identities with the mouse, human, chicken, and carp counterparts. Whole-mount in situ hybridization showed that the lysozyme C gene was expressed in macrophages, as its expression was co-localized with the known myeloid lineage markers L-plastin and PU.1. At 20 hours postfertilization (hpf), most of the lysozyme C positive cells were localized in the yolksac and head mesenchyme but not in the intermediate cell mass, supporting the notion that the primitive macrophage originated from the yolksac (Development 126 (1999) 3735). At 36hpf, the lysozyme C positive cells scattered within the head and yolksac, and began to appear in the caudal part of axial vein. By 6 days postfertilization (dpf), the lysozyme C positive cells accumulated in the kidney where hematopoiesis had been indicated to take place after 4dpf (Dev. Dyn. 214 (1999) 323). Taken together, our results demonstrate that the lysozyme C gene is specifically expressed in myeloid lineage, suggesting that it could serve as an excellent marker for genetic screening of both primitive and definitive myeloid lineage development in zebrafish. PMID- 11900977 TI - Cloning and expression of Ventrhoid, a novel vertebrate homologue of the Drosophila EGF pathway gene rhomboid. AB - In Drosophila melanogaster, the seven-pass transmembrane protein Rhomboid (Rho) is a crucial positive modulator of EGF signaling playing a substantial role in patterning of the ventral neuroectoderm and fate specification of neuroblasts. Here, we describe the cloning and expression pattern of Ventrhoid (Vrho), the novel evolutionarily conserved vertebrate cDNA related to fruit fly rho. Most importantly, like rho in Drosophila, Vrho is also expressed in a spatially restricted manner. Vrho expression is most prominent along the developing ventral neural tube, and is also detectable in the ventral forebrain, prospective diencephalon, otic vesicles, mandibular arches, cranial sensory placodes, last formed pair of somites and hindgut in midgestational mouse embryos. PMID- 11900978 TI - Expression patterns of fibroblast growth factors-18 and -20 in mouse embryos is suggestive of novel roles in calvarial and limb development. AB - The normal development of calvarial bones and sutures critically relies on proper signalling through Fgf receptors, but the source and identity of cognate ligands have remained unknown. Reverse transcriptase polymerase chain reaction analysis in this study shows that a broad range of Fgf ligands are expressed in the coronal sutures separating the parietal and frontal bones. Analysis by whole mount in situ hybridization further reveals distinct expression patterns for Fgf 18, Fgf-20, and by comparison, Fgf-9, in the calvaria, and Fgfs-20 and -9 in the developing limbs, suggestive of their role in proliferation, differentiation and apoptosis. PMID- 11900979 TI - The proneural genes NEUROD1 and NEUROD2 are expressed during human trophoblast invasion. AB - During early human pregnancy, extravillous trophoblast cells invade the maternal tissue of the uterus in a way similar to invasion by cancer cells. However, the process of trophoblast invasion is regulated in a time and place restricted way, in contrast to cancer invasion. We screened first trimester placental tissue enriched by extravillous invasive trophoblasts for the expression of proneural basic helix-loop-helix (bHLH) transcription factors, which are important controllers of cell fate. Surprisingly, the presence of NEUROD1, NEUROD2 and ATH2 transcripts was found by reverse transcriptase polymerase chain reaction (RT-PCR) analysis in first trimester placentabed. Of these genes, the proneural genes NEUROD1 and NEUROD2 are expressed in different subsets of invasive trophoblasts. NEUROD1 expression is found in interstitial and endovascular invasive cells, while NEUROD2 expression is observed mainly in endovascular invasive cells, respectively. These data suggest that in addition to the involvement of proneural genes in neuron, neurendocrine and pancreas differentiation, these genes are involved in trophoblast differentation during progression of invasion. PMID- 11900980 TI - Identification of PGC7, a new gene expressed specifically in preimplantation embryos and germ cells. AB - The gene expression patterns of primordial germ cells (PGCs) and embryonic stem cells were analyzed by a modified serial analysis of gene expression. During the process, we cloned a novel gene, PGC7, which was preferentially expressed in PGCs. Immunohistochemical analysis revealed that PGC7 was specifically expressed in early pre-implantation embryos, PGCs and oocytes. These results suggest that PGC7 might play an important role in the development of PGCs and oocytes. PMID- 11900981 TI - Molecular cloning and expression of the chromatin insulator protein CTCF in Xenopus laevis. AB - The zinc finger protein CTCF has been shown to mediate multiple functions connected to gene repression. Transcriptional inhibition as well as enhancer blocking and chromatin insulation are documented for CTCF in men, mice and chickens. Additionally, hCTCF has been linked to epigenetics and disease. In line with these basic cellular functions, CTCF has been found to be expressed in every cell type and adult tissue tested and has thus been deemed an ubiquitous protein. Here, we report the identification of the CTCF homologue from Xenopus and the analysis of the spatio-temporal expression of xCTCF during embryogenesis. Within the DNA binding domain, xCTCF is virtually identical to other identified vertebrate CTCF proteins. Homology also extends to other conserved regions that are important for CTCF function. Although xCTCF mRNA is present during all stages of early Xenopus development, a remarkable increase in expression is observed in neuronal tissues. Early in development, xCTCF is highly expressed in the neural plate and later in the neural tube and developing brain. By tailbud stage, elevated expression is also seen in the developing sensory organs of the head. This is the first detailed description of the expression pattern of a vertebrate insulator protein during embryogenesis. PMID- 11900983 TI - Ran GTPase expression during early development of the mouse embryo. AB - Ran is a small GTP-binding protein involved in several essential roles for cell viability. This observation implies Ran might be ubiquitously expressed during development. However, Ran shows a differentiated expression pattern that is restricted to specific tissues from embryo to adult. At early embryonic stages of mouse development we found persistent Ran expression in proliferating neural tissue, neural crest derived dorsal root ganglions and sensory pits. We also showed an accumulation of Ran transcripts in main embryonic haematopoietic tissues: blood islands first and then hepatic bud. In advanced stages of development Ran is also expressed in other tissues showing a high cell turnover. PMID- 11900982 TI - Expression of the helix-loop-helix gene id3 in the zebrafish embryo. AB - Proteins of the Extramacrochaetae and Id subfamily of Helix-Loop-Helix (HLH) proteins are negative regulators of bHLH transcription factors. We cloned a cDNA from zebrafish which encodes a member of the id3 subfamily. High levels of transcripts accumulated in the germ ring and in the embryonic shield. Towards the end of gastrulation, Id3 was highly expressed in the anterior prechordal plate and hypoblast. At later stages, id3 expression was turned on and off in a large variety of tissues within short periods of time. These include the lateral mesoderm, the cornea, the lens, the brain, the neural crest, the retina and the fins. PMID- 11900984 TI - The expression of gbx-2 during zebrafish embryogenesis. AB - Gbx-2 is a homeobox gene essential for normal development of the midbrain and the anterior hindbrain. Zebrafish gbx-2 shares an overall similarity of 67.8, 68.1, 60.6 and 66.5% in amino acid sequence to human, mouse, chick and Xenopus Gbx-2, respectively. The expression of zebrafish gbx-2 is initially, before the completion of epiboly, restricted to the prospective posterior midbrain. The expression remains detectable until the end of pharyngula period. The gbx-2 mRNA is also detected in the otic vesicles, the dorsoposterior telencephalon, the rostral branchial arches, the pronephric duct and median fin fold. PMID- 11900986 TI - The roles of IFN gamma in protection against tumor development and cancer immunoediting. AB - Interferon-gamma (IFN gamma) is a cytokine that plays physiologically important roles in promoting innate and adaptive immune responses. The absence of IFN gamma production or cellular responsiveness in humans and experimental animals significantly predisposes the host to microbial infection, a result that validates the physiologic importance of this cytokine in preventing infectious disease. Recently, an additional role for IFN gamma in preventing development of primary and transplanted tumors has been identified. Although there now appears to be a consensus that IFN gamma promotes host responses to tumors, the mechanisms by which this cytokine achieves its effects remain unclear. In this review, we briefly discuss key issues of the molecular cell biology of IFN gamma and its receptor that are most relevant to IFN gamma-dependent anti-tumor effects and then focus on the data implicating IFN gamma as a critical immune system component that regulates tumor development. Potential mechanisms underlying IFN gamma's anti-tumor effects are discussed and a preliminary integrative model of IFN gamma's actions on tumors is proposed. Finally, the capacity of IFN gamma and lymphocytes to not only provide protection against tumor development but also to sculpt the immunogenic phenotype of tumors that develop in an immunocompetent host is presented and introduced as a "cancer immunoediting" process. PMID- 11900987 TI - Endogenous type I interferons as a defense against tumors. AB - We have reviewed the experimental results which indicate that endogenous type I interferon (IFN) present either constitutively or possibly induced by the tumor plays an important role in limiting the development of transplantable tumors in mice. Thus, treatment with potent polyclonal neutralizing antibodies to IFN alpha/beta markedly enhanced the subcutaneous growth, invasiveness and metastases of xenogeneic tumor cells (uninfected or infected with RNA or DNA viruses) in athymic nude mice; enhanced the intraperitoneal transplantability of six different syngeneic murine tumors in three strains of immunocompetent mice; and completely abrogated the resistance of allogeneic C57Bl/6 (H-2(b)) or C3H (H 2(k)) mice to the multiplication of Friend erythroleukemia cells (H-2(d)) in the liver and spleen resulting in the death of most mice. The mechanisms by which mice respond to the injection of relatively few tumor cells appear to be multiple, to depend on the site of tumor growth, to occur early and prior to an immunologic response. Endogenous type I IFN appears to constitute an essential component of these defense mechanisms enabling the host to restrict tumor growth. PMID- 11900988 TI - Interferon-alpha in tumor immunity and immunotherapy. AB - Interferon-alpha (IFN-alpha) is a pleiotropic cytokine belonging to type I IFN, currently used in cancer patients. Early studies in mouse tumor models have shown the importance of host immune mechanisms in the generation of a long-lasting antitumor response to type I IFN. Recent studies have underscored new immunomodulatory effects of IFN-alpha, including activities on T and dendritic cells, which may explain IFN-induced tumor immunity. Reports on new immune correlates in cancer patients responding to IFN-alpha represent additional evidence on the importance of the interactions of IFN-alpha with the immune system for the generation of durable antitumor response. This knowledge, together with results from studies on genetically modified tumor cells expressing IFN alpha, suggest novel strategies for using these cytokines in cancer immunotherapy and in particular the use of IFN-alpha as an immune adjuvant for the development of cancer vaccines. PMID- 11900989 TI - Tumor necrosis factor or tumor promoting factor? PMID- 11900990 TI - Chemokines in cancer. AB - Chemokines participate, by regulating cell trafficking and controlling angiogenesis, in the host response during infection and inflammation. Most of these mechanisms are also operating in cancer. The stimulation of angiogenesis and tumor growth--directly or indirectly through the recruitment of tumor associated macrophages--are typical situations where chemokines promote tumor development. On the other hand, chemokines could be used to the benefit of cancer patients as they act in the recruitment of dendritic cells (DC) or/and effector cells or for their angiostatic properties. However, chemokine-mediated recruitment of immature DC within tumors, due to factors produced by the tumor milieu, could lead to the induction of immune tolerance and, therefore, novel strategies to eradicate tumors based on chemokines should attempt to avoid this risk. PMID- 11900991 TI - Interleukin-12 in anti-tumor immunity and immunotherapy. AB - Interleukin-12 (IL-12) has an essential role in the interaction between the innate and adaptive arms of immunity by regulating inflammatory responses, innate resistance to infection, and adaptive immunity. Endogenous IL-12 is required for resistance to many pathogens and to transplantable and chemically induced tumors. In experimental tumor models, recombinant IL-12 treatment has a dramatic anti tumor effect on transplantable tumors, on chemically induced tumors, and in tumors arising spontaneously in genetically modified mice. IL-12 utilizes effector mechanisms of both innate resistance and adaptive immunity to mediate anti-tumor resistance. IFN-gamma and a cascade of other secondary and tertiary pro-inflammatory cytokines induced by IL-12 have a direct toxic effect on the tumor cells or may activate potent anti-angiogenic mechanisms. The stimulating activity of IL-12 on antigen-specific immunity relies mostly on its ability to determine or augment Th1 and cytotoxic T lymphocyte responses. Because of this ability, IL-12 has a potent adjuvant activity in cancer and other vaccines. The promising data obtained in the pre-clinical models of anti-tumor immunotherapy have raised much hope that IL-12 could be a powerful therapeutic agent against cancer. However, excessive clinical toxicity and modest clinical response observed in the clinical trials point to the necessity to plan protocols that minimize toxicity without affecting the anti-tumor effect of IL-12. PMID- 11900992 TI - Interleukin-2 and interleukin-15: immunotherapy for cancer. AB - Interleukin (IL)-2 and IL-15 are two cytokine growth factors that regulate lymphocyte function and homeostasis. Early clinical interest in the use of IL-2 in the immunotherapy of renal cell carcinoma and malignant melanoma demonstrated the first efficacy for cytokine monotherapy in the treatment of neoplastic disease. Advances in our understanding of the cellular and molecular biology of IL-2 and its receptor complex have provided rationale to better utilize IL-2 to expand and activate immune effectors in patients with cancer. Exciting new developments in monoclonal antibodies recognizing tumor targets and tumor vaccines have provided new avenues to combine with IL-2 therapy in cancer patients. IL-15, initially thought to mediate similar biological effects as IL-2, has been shown to have unique properties in basic and pre-clinical studies that may be of benefit in the immunotherapy of cancer. This review first summarizes the differences between IL-2 and IL-15 and highlights that better understanding of normal physiology creates new ideas for the immunotherapy of cancer. The application of high, intermediate, and low/ultra low dose IL-2 therapy in clinical trials of cancer patients is discussed, along with new avenues for its use in neoplastic diseases. The growing basic and pre-clinical evidence demonstrating that IL-15 may be useful in immunotherapy approaches to cancer is also presented. PMID- 11900993 TI - GM-CSF-based cellular vaccines: a review of the clinical experience. AB - Immunotherapy is playing an increasing role in the treatment of many cancers. The recent advances in antibody therapy gives much optimism that both passive (antibody therapy) as well as active (vaccine therapy) immunotherapeutic interventions will acquire an increasing presence in oncology. Granulocyte macrophage-colony stimulation factor (GM-CSF)-based vaccines have now been tested in several diseases in a variety of formulations. The success and broad applicability of such an approach rests on the development of an ideal vaccine formulation administered in the appropriate clinical context. This review summarizes the results from the clinical trials performed to date and discusses the future directions of GM-CSF-based cellular vaccine strategies aimed at maximizing the therapeutic benefit. PMID- 11900994 TI - Pharmacological concentrations of the HMG-CoA reductase inhibitor lovastatin decrease the formation of the Alzheimer beta-amyloid peptide in vitro and in patients. AB - Epidemiological studies demonstrate that hypercholesterolemia is a risk factor for Alzheimer's disease (AD). As the generation and accumulation of the beta amyloid peptide (Abeta) in the brain appears to be significant for the initiation and progression of AD, it is possible that cholesterol levels regulate Abeta formation and/or clearance. To test the effects of altering cholesterol on Abeta formation, we incubated cells with or without lovastatin acid, the active metabolite of the HMG-CoA reductase inhibitor lovastatin, and measured the fraction of Abeta formed from its precursor under each condition. We observed that treatment with lovastatin acid led to a profound decrease in the levels of Abeta formed. This effect was observed at concentrations of 0.05-5 microM, ranges where this compound is effective at inhibiting HMG-CoA reductase. To examine the effects of lovastatin on Abeta in vivo, human subjects who had elevated low density lipoprotein cholesterol were treated during a double-blind, randomized study with 10-60-mg once-daily doses of a controlled-release formulation of lovastatin, or matching placebo. Serum Abeta concentrations were measured before and after up to 3 months of treatment. Mean and median changes from baseline in serum Abeta concentrations showed a significant (p < 0.0348), dose-dependent decrease. Differences between the 40- and 60-mg dose groups and placebo were statistically significant (Dunnett's p< 0.05). Our results suggest a mechanism by which hypercholesterolemia may increase risk for AD and indicate that lovastatin reduces Abeta formation and may thereby be effective in delaying the onset and/or slowing the progression of AD. PMID- 11900995 TI - Gliomatosis cerebri has normal relative blood volume: really?! Who cares? Should you? PMID- 11900996 TI - The emergence of time-resolved contrast-enhanced MR imaging for intracranial angiography. PMID- 11900997 TI - Achieving gross total resection of brain tumors: intraoperative MR imaging can make a big difference. PMID- 11900998 TI - Dynamic contrast-enhanced T2*-weighted MR imaging of gliomatosis cerebri. AB - BACKGROUND AND PURPOSE: MR imaging characteristics of gliomatosis cerebri reiterate the diffuse nature of this tumor but are nonspecific and thus may pose a diagnostic challenge. Because perfusion MR imaging can provide a physiologic map of the microcirculation, we compared the measured relative cerebral blood volume (rCBV) at perfusion imaging with histopathologic findings in gliomatosis cerebri. MR spectroscopic findings were also reviewed. METHODS: Retrospective analysis was performed of conventional and perfusion MR images from seven patients with proved gliomatosis cerebri. The conventional MR images were evaluated for the presence or absence of contrast enhancement, necrosis, and extent of T2-weighted signal intensity abnormality. Dynamic contrast-enhanced T2* weighted gradient-echo echo-planar images were acquired during the first pass of a bolus injection of gadopentetate dimeglumine. The rCBV was calculated by using nondiffusible tracer kinetics and expressed relative to normal-appearing white matter. Pathologic findings were reviewed in all patients and compared with the MR perfusion data. Multivoxel 2D chemical shift imaging proton MR spectroscopic data were available for three patients and single-voxel data for one patient. RESULTS: Conventional MR images showed diffuse abnormality in all cases and absence of contrast enhancement in all but one case. Average rCBV range was 0.75 1.26 (mean, 1.02 +/- 0.42 [SD]). MR spectroscopic data revealed spectra consistent with presence of tumoral disease. Histopathologic review showed absence of vascular hyperplasia in all specimens. CONCLUSION: The low MR rCBV measurements of gliomatosis cerebri are in concordance with the lack of vascular hyperplasia found at histopathologic examination; thus, perfusion MR imaging provides useful adjunctive information that is not available from conventional MR imaging techniques. PMID- 11900999 TI - Estimation of tumor volume with fuzzy-connectedness segmentation of MR images. AB - BACKGROUND AND PURPOSE: Reproducible measurements of brain tumor volume are helpful in evaluating the response to therapy and the need for changing treatment plans. Our purpose was to adapt the fuzzy-connectedness segmentation technique to measure tumor volume. This technique requires only limited operator interaction. METHODS: Routine postoperative brain MR imaging was performed in 19 patients with primary malignant gliomas of the brain. Segmentation was performed on axial and coronal gadolinium-enhanced and axial fluid-attenuated inversion recovery (FLAIR) images by using a fuzzy-connectedness algorithm, and tumor volumes were generated. Operator interaction was limited to selecting representative seed points within the tumor and, if necessary, editing the segmented image to include or exclude improperly classified regions. RESULTS: Measurements of tumor volume were highly reproducible when they were obtained with no editing; intraobserver coefficients of variation were 0.15-0.37% and 0.29-0.38%, respectively, for enhanced images and FLAIR images. Editing consistently produced smaller volumes, at the cost of greater variability in volume measurements. Coefficients of variation for volumes with editing ranged from 0.2% to 1.3%. CONCLUSIONS: Fuzzy connected segmentation permits rapid, reliable, consistent and highly reproducible measurement of tumor volume from MR images with limited operator interaction. PMID- 11901000 TI - Primary sellar lymphoma: radiologic and pathologic findings in two patients. AB - We describe the clinical, MR imaging, and pathologic findings in two cases of primary sellar lymphoma in immunocompetent individuals. Both patients had hypopituitarism, and one patient had a sixth cranial nerve palsy. MR images depicted sellar and suprasellar masses in both patients, and extension into the cavernous and sphenoid sinuses was also present in one patient. The pathologic diagnosis in both cases was large B cell lymphoma. PMID- 11901001 TI - MR imaging characteristics of amyloid deposits in pituitary adenoma. AB - Although cases of pituitary adenomas containing amyloid deposits have been described in the literature, to our knowledge this is the first report to describe MR imaging characteristics of a pituitary adenoma containing almost entirely amyloid tissue. PMID- 11901002 TI - Whole-brain N-acetylaspartate concentration: correlation with T2-weighted lesion volume and expanded disability status scale score in cases of relapsing-remitting multiple sclerosis. AB - BACKGROUND AND PURPOSE: The T2-weighted MR imaging total lesion volume and Expanded Disability Status Scale (EDSS) score are two common measures of relapsing-remitting multiple sclerosis disability and pathologic abnormality. Because the whole-brain N-acetylaspartate concentration is considered to be a new marker of the disease burden, the purpose of this study was to evaluate the relationship among these three measures. METHODS: The whole-brain N acetylaspartate concentration and T2-weighted lesion volume were quantified by using MR imaging and proton MR spectroscopy in 49 patients with relapsing remitting multiple sclerosis (36 female and 13 male patients; average age, 39 years; age range, 24-55 years; average EDSS score, 2; range of EDSS scores, 0-6). Correlations among whole-brain N-acetylaspartate concentrations, T2-weighted lesion volumes, and EDSS scores were obtained. RESULTS: No correlation was found between whole-brain N-acetylaspartate levels and either T2-weighted lesion volumes or EDSS scores. A weak correlation was found between the EDSS scores and T2-weighted lesion volumes (P =.043, r(s) = 0.292). CONCLUSION: Despite the lack of correlation between whole-brain N-acetylaspartate concentration and the clinical disability reflected in the EDSS score, only the former evaluates the global neuronal cell disease in the entire brain, including those lesions that are occult to conventional imaging techniques. PMID- 11901003 TI - Sandlike appearance of Virchow-Robin spaces in early multiple sclerosis: a novel neuroradiologic marker. AB - BACKGROUND AND PURPOSE: The distinctive hyperintensity of multiple sclerosis (MS) lesions on T2-weighted brain MR images is well recognized. However, Virchow-Robin spaces (VRSs), especially in early MS, have not been described. Our purpose was to determine the frequency of VRSs in recent-onset MS. METHODS: Brain MR imaging was performed in 71 patients (mean age, 26.8 years; range, 20-41 years; 47 women, 24 men) within 3 months of MS onset. Proton density-, T2-, and T1-weighted images were obtained. Age-and sex-matched control subjects (mean age, 27.2 years; range, 22-41 years; 38 women, 22 men) who underwent brain MR imaging as a part of headache evaluation, and findings that were interpreted as normal served as controls. On high-convexity images (axial sections above the upper corpus callosum border), VRSs were identified as small (<2-mm diameter) sandlike areas isointense to CSF. VRSs were graded 0-3. RESULTS: VRSs were visualized in high convexity white matter in 55% of patients and 7% of control subjects (P <.001). In patients, 15% of VRSs were grade 1 (fewer than four), 23% were grade 2 (four to seven), and 62% were grade 3 (more than seven). In control subjects, all identified VRSs were grade 1. Among patients with and those without VRSs, age at onset, neurologic disability, and specific functional system involvement or mono- versus polysymptomatic involvement at onset did not differ. CONCLUSION: VRSs were more frequent in patients with recent-onset MS than in control subjects. The sandlike appearance of VRSs may be a neuroradiologic marker that reflects early inflammatory changes in MS. PMID- 11901005 TI - Identification of a by-product of nitric oxide synthase activity in human acute brain injury with in vivo proton magnetic resonance spectroscopy. AB - BACKGROUND AND PURPOSE: Laboratory studies have been used to identify nitric oxide as a notable mediator in neuronal death after acute brain injury. To our knowledge, this has not previously been confirmed with in vivo study in humans. Our purpose was to seek in vivo evidence for the induction of nitric oxide synthase (NOS) in human acute brain injury by using proton MR spectroscopy. METHODS: In vitro proton MR spectra were obtained in neural extracts from 30 human cadavers, and in vivo spectra were obtained in 20 patients with acute brain injury and in a similar number of control subjects. RESULTS: We identified a unique peak at 3.15 ppm by using in vivo proton MR spectroscopy in eight of 20 patients with acute brain injury but not in 20 healthy volunteers (P <.002). On the basis of in vitro data, we have tentatively assigned this peak to citrulline, a NOS by-product. CONCLUSION: To our knowledge, our findings suggest, for the first time, that excitotoxicity may occur in human acute brain injury. Confirmation with the acquisition of spectra in very early acute cerebral injury would provide a rationale for the use of neuroprotective agents in these conditions, as well as a new noninvasive method for quantification. PMID- 11901004 TI - Quantitative measurement of regional cerebral blood flow with flow-sensitive alternating inversion recovery imaging: comparison with [iodine 123] iodoamphetamin single photon emission CT. AB - BACKGROUND AND PURPOSE: Flow-sensitive alternating inversion recovery (FAIR) MR imaging is a technique for depicting cerebral perfusion without contrast enhancement. Our purpose was to determine whether quantification at FAIR imaging can be used to assess regional cerebral blood flow (rCBF) in a manner similar to [iodine 123]-iodoamphetamin ((123)I-IMP) single photon emission CT (SPECT). METHODS: Nine patients with internal carotid or major cerebral arterial stenosis underwent (123)I-IMP SPECT and FAIR imaging (single section, different TIs, 1.5 T) at rest and after acetazolamide (Diamox) stress. FAIR and (123)I-IMP rCBF values were compared and correlated. Receiver operating characteristic analysis was conducted to detect hypoperfused segments on FAIR images. RESULTS: rCBF values of normally perfused segments were 41.53 and 51.91 mL/100 g/min for pre- and post-acetazolamide (123)I-IMP studies, respectively. Corresponding values for pre- and post-acetazolamide FAIR images, respectively, were 46.64 and 59.60 mL/100 g/min with a TI of 1200 milliseconds and 53.23 and 68.17 mL/100 g/min with a TI of 1400 milliseconds. (123)I-IMP and FAIR results were significantly correlated, with both pre- and post-acetazolamide images. Sensitivity (86%) in detecting hypoperfused segments was significantly higher with post-acetazolamide images (TI, 1400 milliseconds), and specificity (82-85%) and accuracy (80-82%) were higher with all pre- and post-acetazolamide images (all TIs). CONCLUSIONS: The significant correlation, high specificity and accuracy in detecting hypoperfused segments, similar increases in flow on both post-acetazolamide images, and absence of the need for contrast enhancement suggest that FAIR imaging, like nuclear medicine study, is complementary to routine MR imaging in the assessment of cerebral perfusion. PMID- 11901006 TI - Influence of imaging parameters on high-intensity cerebrospinal fluid artifacts in fast-FLAIR MR imaging. AB - BACKGROUND AND PURPOSE: High-intensity CSF artifacts at the basal cisterns on MR images are often seen when a fast fluid-attenuated inversion recovery (FLAIR) technique is used. We investigated the influences of four optional fast-FLAIR sequence parameters on the high-intensity CSF artifacts. METHODS: A total of 377 patients (age range, 1 week to 91 years; mean 40.6 years; 186 female, 191 male) were examined with axial fast-FLAIR images obtained (TR/TE(eff)/TI, 8800/133/2200) with a 1.5-T system during 6 months. The effects of the optional addition of inferior inflow saturation (thickness, 80 mm), section flow compensation, and tailored radiofrequency (TRF) pulses, plus the choice of interleaving acquisition factors of 2 or 3, were evaluated for the presence of high-intensity CSF artifacts on the fast-FLAIR images. Two radiologists independently reviewed the fast-FLAIR images in 76 patients; afterward, a single observer reviewed the remainder of the images. RESULTS: The interobserver agreement rate in 76 cases was more than 90%. The use of TRF and/or three interleaving acquisitions resulted in a substantial reduction in the incidence of high-intensity CSF artifacts from about 80% to 40% (P <.05, two-sample two-sided Z test). Inferior inflow saturation and section flow compensation did not significantly improve image quality (P >.05). The results were consistent with the image quality ranking obtained in five healthy volunteers. CONCLUSION: The appropriate choice of sequence parameters in fast-FLAIR imaging reduces the incidence of high-intensity CSF artifacts that are frequently encountered in the presence of rapid CSF flow. PMID- 11901007 TI - Visualization of subdural strip and grid electrodes using curvilinear reformatting of 3D MR imaging data sets. AB - Curvilinear reformatting of 3D MR imaging data sets was used to visualize the position of subdural strip and grid electrodes relative to the underlying cerebral cortex in patients with epilepsy who were undergoing invasive electroencephalographic recordings. The contour of the cortical surface was delineated interactively, and topographical relationships among surface gyration, cortical lesions, and subdural electrodes were investigated by using serial convex planes parallel to the cortical surface. Electrode contacts could be marked and their positions projected to underlying areas at different depths. This method is apt for routine purposes and allows electrode positions to be displayed with respect to cortical and subcortical regions of interest. PMID- 11901008 TI - Dural arteriovenous fistulae: noninvasive diagnosis with dynamic MR digital subtraction angiography. AB - MR digital subtraction angiography (DSA) is a new diagnostic tool capable of producing dynamic images of the cerebral circulation with the injection of gadopentetate dimeglumine into a peripheral vein. Previous reports have concentrated on its potential as a noninvasive technique for the study of pial arteriovenous malformations. In this report, we present our early findings with MR DSA in the evaluation of intracranial dural arteriovenous fistulae. PMID- 11901009 TI - Diagnostic pitfall: atypical cerebral venous drainage via the vertebral venous system. AB - We report a case of atypical cerebral venous drainage in a 38-year-old woman with symptoms of benign paroxysmal positional vertigo. Thrombosis of the left internal jugular vein and sigmoid sinus was suspected on the basis of spin-echo and time of-flight MR findings, but multisection CT angiograms showed a patent sigmoid sinus and predominant drainage via the emissary veins toward the vertebral plexus, with only a minor contribution of the jugular veins. This case illustrates the variability of the venous anatomy in the craniocervical region. PMID- 11901010 TI - Reproducibility analysis of a new objective method for measuring arteriovenous malformation nidus size at angiography. AB - BACKGROUND AND PURPOSE: Currently, no specific method exists to measure arteriovenous malformation (AVM) nidus size, a requirement in assessing the success of treatment. Additionally, the commonly used evaluation provides only a linear one-dimensional measurement of this three-dimensional entity. The purpose of this study was to devise an improved method for measuring AVM nidus size, an irregularly shaped radiologic entity, that provides objective and reproducible results. METHODS: The procedure involved digitizing angiograms obtained before and after treatment, making the gray scale uniform, printing images on standard bond paper, delineating the nidus area, measuring the nidus area with a polar planimeter, and finally, correcting for geometric magnification. Three observers made the measurements. The corrected nidus areas were tabulated, and the mean, standard deviation, interobserver variability, and confidence intervals (CIs) were calculated. RESULTS: On both anteroposterior and lateral views, the Kendall coefficient of concordance (a measure of interobserver variability) was equal to 0.97, signifying excellent agreement. Additionally, these values were within the 95% CIs; this result showed that they were unlikely the result of chance. CONCLUSION: Precise measurements of an AVM nidus are required to properly analyze changes in the lesion after endovascular embolization (ie, to evaluate treatment success). Because of the irregular contours of an AVM nidus, measuring an area with planimetry, rather than with the usual linear dimensions, should yield more exact results. PMID- 11901011 TI - Tailored cognitive testing with provocative amobarbital injection preceding AVM embolization. AB - BACKGROUND AND PURPOSE: Transarterial embolization of cerebral arteriovenous malformations (AVMs) has been associated with postprocedural neurologic complications in 7-39% of patients. We sought to determine whether a method of targeted neurologic and cognitive testing during AVM embolization reduces the incidence of focal cognitive and other neurologic deficits associated with the procedure. METHODS: A cognitive neurologist extensively examined 12 patients prior to AVM embolization. In each patient, a battery of tests tailored to their specific abilities was developed by using stimuli selected from standard and experimental cognitive tests to probe specific brain regions related to the location of the AVM. In each feeder vessel to be embolized, a 50-mg bolus of sodium amobarbital was superselectively administered through a microcatheter; this was followed immediately by neurologic and cognitive testing with the tailored battery. After testing, the position of the microcatheter tip was checked with fluoroscopy. If the provocative test results were negative, the evaluated feeder was embolized with N-butyl cyanoacrylate glue. RESULTS: Although results with 27 of 29 provocative amobarbital injections were negative, results with two injections in two different individuals revealed cognitive deficits during tailored provocative testing. In both, the evoked deficits resolved with dissipation of the amobarbital effect; the feeder vessels were not embolized. Neurologic and cognitive evaluation after each of 27 embolizations revealed no major or minor deficits. CONCLUSION: In our experience, provocative amobarbital testing prior to AVM embolization was helpful in identifying vascular territories where embolization may lead to neurologic and cognitive deficits. PMID- 11901012 TI - Liquid 2-poly-hydroxyethyl-methacrylate embolization of experimental arteriovenous malformations: feasibility study. AB - BACKGROUND AND PURPOSE: We investigated the use of 2-poly-hydroxyethyl methacrylate (2-P-HEMA) as an embolic agent in swine arteriovenous malformations (AVMs). METHODS: In seven mini swine, experimental AVMs were created surgically. The aim of treatment was complete embolization of the nidus compartment filled by the feeding artery, without brain embolization. Six animals received pure liquid 2-P-HEMA, and one, 50% 2-P-HEMA. For radiopacity, liquid 2-P-HEMA was mixed with tungsten powder. Six animals underwent angiographic follow-up within 5-8 mo (mean, 6.5 mo). Evaluation criteria were controllability, procedural reproducibility, and duration of the nidus occlusion. To detect complications, brain MR imaging and CT were performed. Histopathologic studies were performed to prove occlusion and assess histopathologic responses. RESULTS: 2-P-HEMA was easily injected through microcatheters, with a reproducible technique. Because of the radiopacity of the mixture, deep nidus penetration was controlled with fluoroscopy and confirmed with CT and histopathologic examination. In five AVMs embolized with pure 2-P-HEMA, feeder obliteration was long term. One animal had vasospasm during embolization, and complete obliteration of the main feeder was maintained for 3 mo, but partial recanalization developed 2 mo later. One animal receiving pure 2-P-HEMA had an infarction. In the animal embolized with 50% 2-P HEMA, angiography and CT revealed embolic material in the circle of Willis; the animal died after embolization. No marked inflammatory reaction in the vessel wall or perivascular tissue was observed in the embolized AVMs. CONCLUSION: Experimental AVM embolization with pure 2-P-HEMA, made radiopaque with tungsten, is technically feasible in swine. Because of its properties, 2-P-HEMA has great potential as a therapeutic embolic agent. PMID- 11901013 TI - Angioplasty and stent placement in intracranial atherosclerotic stenoses and dissections. AB - BACKGROUND AND PURPOSE: Stent placement has been shown to increase the safety and effectiveness of balloon angioplasty in cervical carotid disease. Here, the authors investigated the feasibility, safety, and short-term outcome of stent assisted angioplasty for the treatment of intracranial stenoses. METHODS: Thirty four patients (age range, 12-77 years; mean age, 54 years) with symptomatic intracranial atherosclerotic lesions and dissections that produced stenosis of more than 50% were selected and treated with stents. Eighteen lesions (53%) were located in the anterior circulation, and 16 (47%) were in the vertebrobasilar complex. The mean stenosis was 75%. RESULTS: At follow up, 21 patients (62%) improved clinically, 11 (32%) remained stable, and the condition of two patients (6%) deteriorated. In all patients, the angiographic degree of stenosis was reduced to less than 30%. In 10 patients (29%), two or more stents were implanted: Two stents were implanted in six patients, and three, in four patients. The transient procedural morbidity rate was 12%, and the transient neurologic morbidity rate was 6%. One patient had hemorrhagic transformation due to reperfusion and died, and another patient had a massive myocardial infarction after 5 months. Twenty patients were followed up with angiography for at least 6 months, and none required repeat angioplasty. CONCLUSION: Endovascular revascularization of intracranial arteries by means of stent-assisted angioplasty is technically feasible, effective, and safe in selected patients. PMID- 11901014 TI - Stent-assisted coil placement in a wide-necked persistent trigeminal artery aneurysm with jailing of the trigeminal artery: a case report. AB - We report the successful stent placement and secondary coil placement in a wide necked persistent trigeminal artery (TA) aneurysm, in which the internal carotid artery (ICA) was preserved and the TA "jailed." Nine-month follow-up angiography revealed a completely obliterated aneurysm and patent parent ICA and TA. MR images obtained at 16-mo follow-up showed no infarction of the brain in the distribution of the jailed TA. The risks of stent jailing and possible solutions are discussed. PMID- 11901015 TI - Treatment of iatrogenic internal carotid artery laceration and carotid cavernous fistula with endovascular stent-graft placement. AB - The risk of fatal injury of the internal carotid artery (ICA) and surrounding anatomy during transsphenoidal surgery for pituitary adenoma is the most severe potential complication associated with this particular approach. We present a case in which iatrogenic injury to a patient's ICA and resultant carotid cavernous fistula and massive hemorrhage was successfully managed with the emergency placement of an endovascular stent-graft. Both findings in the relevant literature and practical considerations concerning both stent-grafts and other more commonly used options for the treatment of iatrogenic ICA injury are discussed. PMID- 11901016 TI - Intraarterially administered abciximab as an adjuvant thrombolytic therapy: report of three cases. AB - The intravenous administration of abciximab can be used as an adjuvant therapy to facilitate thrombolysis for acute cerebrovascular occlusion. However, to our knowledge, the intraaterial administration of abciximab has not been reported. We recently treated three patients with acute thrombosis of the cerebral arteries by using an intraarterial infusion of urokinase and abciximab. Even with small doses, we achieved rapid and complete recanalization without complications. We believe that the intraarterial infusion of abciximab may be promising for effective and safe recanalization of acute thrombotic occlusion of cerebral arteries. PMID- 11901017 TI - Recognition and importance of an infraoptic anterior cerebral artery: case report. AB - Although variations of the anterior cerebral artery (ACA)-anterior communicating artery complex are commonly identified on imaging studies, an infraoptic course of the ACA is exceedingly rare. What is believed to be the first case of an infraoptic course of the ACA discovered with MR angiography and further characterized with conventional angiography is presented. The high prevalence of associated aneurysms and the implications for surgical planning make preoperative recognition of this anomaly critical. PMID- 11901018 TI - Bilateral arch origin of the vertebral arteries. AB - A case of bilateral anomalous origins of the vertebral arteries (VAs) is reported. Both VAs arose directly from the aortic arch between the left common carotid artery and the left subclavian artery. The possible embryologic mechanism and clinical importance of this previously unreported variant are reviewed. PMID- 11901019 TI - Using Bayesian tissue classification to improve the accuracy of vestibular schwannoma volume and growth measurement. AB - BACKGROUND AND PURPOSE: True 3D measurements of tumor volume are time-consuming and subject to errors that are particularly pronounced in cases of small tumors. These problems complicate the routine clinical assessment of tumor growth rates. We examined the accuracy of currently available methods of size and growth measurement of vestibular schwannomas compared with that of a novel fast partial volume tissue classification algorithm. METHODS: Sixty-three patients with unilateral sporadic vestibular schwannomas underwent imaging. Thirty-eight of these patients underwent imaging two or more times at approximately 12-month intervals. Contrast-enhanced 3D T1-weighted images were used for all measurements. An experienced radiologist performed standard size estimations, including maximal diameter, elliptical area, perimeter, manually segmented area, intensity thresholded seeding volume, and manually segmented volume. A method for calculating volume was also used, incorporating Bayesian probability statistics to estimate partial volume effects. Manually segmented volume was obtained as a baseline standard measure. A computer-generated phantom exhibiting the intensity and partial volume characteristics of brain tissue, CSF, and intracanalicular vestibular schwannoma tissue was used to measure absolute accuracy of the standard technique and Bayesian partial volume segmentation. RESULTS: The Bayesian partial volume segmentation method showed the highest correlation (R(2) = 0.994) with the standard method, whereas the commonly used method of maximal diameter measurement showed poor correlation (R(2) = 0.732). Accuracy of Bayesian segmentation was shown to be more than twice that of manual segmentation, with an absolute accuracy of 5% (cf, 13%) and a remeasurement accuracy of 70 mm(3) (cf, 150 mm(3)). For the 38 patients who underwent imaging twice, definite tumor growth was shown for 12, potential growth for seven, no growth for 17, and definite shrinkage for two. CONCLUSION: Commonly used methods such as maximal diameter measurements do not provide adequate statistical accuracy with which to monitor tumor growth in patients with small vestibular schwannomas. Bayesian partial volume segmentation provides a more accurate and rapid method of volume and growth estimation. These differences in measurement accuracy translated into a significant improvement in clinical assessment, allowing identification of tumor growth in 10 of 12 cases that appeared to be static in size when manual segmentation techniques are used. The technique is quick to perform and suitable for use in routine clinical practice. PMID- 11901020 TI - Deep extension from carcinoma arising from the gingiva: CT and MR imaging features. AB - BACKGROUND AND PURPOSE: CT and MR imaging are useful for evaluating the extension of carcinomas in the face and neck. We evaluated the involvement by carcinoma arising from the gingiva (ie, gingival cancer) by using CT and MR imaging. METHODS: We retrospectively examined 122 patients with squamous cell carcinoma (SCCA) in the lower (88 patients) and upper (34 patients) gingiva. Extension of SCCA into the spaces of the face and neck was evaluated with CT and MR imaging, and findings were surgically confirmed. RESULTS: Spread into the face and neck spaces occurred in 58% of patients. The buccal space was the most common site of spread, occurring in 42% of the lower and of 47% of the upper gingival cancers. Spread into the masticator space occurred from the lower gingival cancers in the molar region (20%) but not from the anterior region. Masticator space involvement from the upper gingiva was rare (4%). The retromolar triangle and buccal space immediately anterior to the ramus served as a corridor for cancer extension from the lower gingiva into the masticator space. The sublingual space (11%) was a less common site of spread from the lower gingiva. CONCLUSION: Gingival cancers spread into the masticator, buccal, and sublingual spaces depending on the primary sites in the oral cavity. An understanding of the face and neck-space anatomy is important in diagnosing cancer extension in the oral cavity gingiva and in treating patients with such disease. PMID- 11901021 TI - Solitary fibrous tumor of the parapharyngeal space: MR imaging findings. AB - We report the MR imaging findings of a solitary fibrous tumor involving the parapharyngeal space. The tumor was a well-circumscribed solid mass with a lobulated contour. It had the same signal intensity as the muscle on T1-weighted MR images, heterogeneously high signal intensity on T2-weighted images, and homogeneous strong enhancement after the administration of contrast material. It mimicked a tumor originating from the deep lobe of the parotid gland. PMID- 11901022 TI - Neuroradiologic-pathologic correlation in a neurenteric cyst of the clivus. AB - A 28-year-old woman presented with left-sided frontotemporal headache lasting 6 wk. Head CT and MR imaging revealed a clival mass, which was interpreted as a chondrosarcoma. The lesion was removed at endoscopic endonasal surgery; histologic and immunohistochemical findings proved it to be neurenteric cyst. On CT scans, the lesion was lytic, with an intact cortex; it was uniformly hyperintense relative to gray matter on T1-weighted MR images and iso- to hypointense relative to CSF on T2-weighted MR images. PMID- 11901023 TI - MR imaging and MR spectroscopy in rhizomelic chondrodysplasia punctata. AB - A case of rhizomelic chondrodysplasia punctata was investigated with MR imaging of the brain and hydrogen-1 MR spectroscopy of the brain and blood. Areas with abnormal signal hyperintensity on T2-weighted images or hypointensity on T1 weighted images were detected in the subcortical white matter. MR spectroscopy of the brain showed that normal-appearing white matter was characterized by increased levels of mobile lipids and myo-inositol, reduced levels of choline, and the presence of acetate. The importance of these metabolic anomalies is correlated to the deficiency in plasmalogen biosynthesis. PMID- 11901024 TI - Isolated sulfite oxidase deficiency: MR imaging features. AB - Isolated sulfite oxidase deficiency is a rare autosomal inherited disorder of the normal degradation of sulfur-containing amino acids. Premature death in infancy secondary to severe neurologic deterioration is the usual outcome. This article provides an analysis, in temporal form, of brain imaging findings in this disorder. PMID- 11901025 TI - Balamuthia amebic encephalitis: radiographic and pathologic findings. AB - The radiographic findings of two patients (one, a rare survivor) with meningoencephalitis caused by Balamuthia mandrillaris are presented with pathologic correlation. PMID- 11901026 TI - Sturge-Weber disease with repercussion on the prenatal development of the cerebral hemisphere. AB - Sturge-Weber syndrome was diagnosed in a neonate on basis of a characteristic port-wine stain. In the absence of any acute neurologic episode, MR images obtained when the infant was aged 3 months showed a typical pial vascular dysplasia, as well as prominent hypotrophy of the homolateral hemisphere. Areas suggesting the presence of developmental dysplasia of the cerebral mantel were found in association with the typical pial vascular anomaly. The prenatal effect of Sturge-Weber disease on normal brain development may best be explored by using a better evaluation with cerebral imaging shortly after birth. PMID- 11901027 TI - Langerhans cell histiocytosis of the skull complicated with an epidural hematoma. AB - Langerhans cell histiocytosis (LCH) is a rare disorder that affects the pediatric population. LCH complicated with a neurologic deficit due to the presence of epidural involvement is a rare condition. We describe the CT imaging features in a 2-year-old boy who presented with drowsy consciousness resulting from an epidural hematoma caused by spontaneous bleeding in an LCH of the skull. CT is an excellent means of depicting the full extent of bony destruction and the nature of the process. PMID- 11901028 TI - Diffusion-weighted imaging in a patient with vertebral and epidural abscesses. AB - In this report, we describe the appearance of the spinal vertebral body and epidural abscesses in a patient who underwent diffusion-weighted imaging of the spine and CT-guided aspiration of one of the abscesses. The abscesses were hyperintense relative to the surrounding tissues on diffusion-weighted images, and they appeared dark on apparent diffusion coefficient maps; these findings were consistent with those of published reports of the brain and liver. PMID- 11901029 TI - The Fourth Asian-Oceanian Congress of Neuroradiology and Head & Neck Radiology. PMID- 11901030 TI - Joel Swartz. PMID- 11901031 TI - Intracranial hemorrhage and cerebral hyperperfusion syndrome after extracranial carotid artery angioplasty and stent placement. PMID- 11901033 TI - Oral anticoagulation for acute coronary syndromes. PMID- 11901034 TI - Successful treatment of amiodarone-induced thyrotoxicosis. AB - BACKGROUND: Amiodarone-induced thyrotoxicosis (AIT) is a difficult management problem about which there are little published data. We examined whether continuing amiodarone or differentiating AIT into 2 subtypes affected outcome. METHODS AND RESULTS: The type and duration of antithyroid treatment and response were recorded in a consecutive series of 28 cases. Comparisons were made between those in whom amiodarone either was continued or stopped and between those with either possible type 1 or type 2 AIT. Of the 28 cases, 5 had spontaneous resolution of AIT; 23 received carbimazole (CBZ) alone as first-line therapy. Eleven achieved long-term euthyroidism off CBZ or on a maintenance dose. Five became hypothyroid and required long-term thyroxine. Five relapsed after stopping CBZ treatment and were rendered euthyroid with either long-term CBZ (n=3) or radioiodine (n=2). Four were intolerant of CBZ and received propylthiouracil (PTU), with good effect in 3. One was resistant to thionamide alone (CBZ then PTU) and responded to adjunctive steroids. No difference in presentation or outcome was noted between those in whom amiodarone was continued or stopped or between possible type 1 or type 2 AIT. CONCLUSIONS: Continuing amiodarone has no adverse influence on response to treatment of AIT. First-line therapy with a thionamide alone is appropriate in iodine-replete areas, thus avoiding potential complications of other drugs. Differentiating between 2 possible types of AIT does not influence management or outcome. PMID- 11901035 TI - Local expression of interleukin-1 receptor antagonist by plasmid DNA improves mortality and decreases myocardial inflammation in experimental coxsackieviral myocarditis. AB - BACKGROUND: The inflammatory cytokines have an important role in the pathogenesis of viral myocarditis. Inerleukin-1 (IL-1) is one of the major cytokines that modulate the outcome of viral infection. Among the methods for in vivo gene transfer, direct injection of plasmid DNA is one that is simple and feasible. In this study, we expressed human IL-1 receptor antagonist (hIL-1Ra) in the mouse heart by direct injection of a novel plasmid vector and evaluated its effects on coxsackieviral (CVB3) myocarditis. METHODS AND RESULTS: A plasmid vector expressing hIL-1Ra (total 40 microg/mouse) was injected into the heart apex of 8 week-old inbred female Balb/C mice (day 3). On day 0, mice (IL-1Ra-CVB3, n=35) were infected intraperitoneally with 10(4) PFU of CVB3; control mice (pCK-CVB3, n=15) were injected with empty vector on day -3 and infected on day 0. hIL-1Ra was expressed in the heart, reached its peak on day 5, and persisted for 2 weeks. The 14-day survival rate of IL-1Ra-CVB3 was higher (77%) than that of controls (30%, P<0.01). Myocardial virus titers on day 3 were lower in IL-1Ra-CVB3 mice. Myocardial inflammation on day 7 and fibrosis on day 14 were markedly decreased in IL-1Ra-CVB3. CONCLUSION: These results showed that direct injection of the expression plasmid vector into the heart was an effective method to transfer the cytokine gene in vivo, and expressed IL-1Ra in the heart can modulate the deleterious effect of the host immune response in viral myocarditis. PMID- 11901036 TI - Catheter-based endomyocardial injection with real-time magnetic resonance imaging. AB - BACKGROUND: We tested the feasibility of targeted left ventricular (LV) mural injection using real-time MRI (rtMRI). METHODS AND RESULTS: A 1.5T MRI scanner was customized with a fast reconstruction engine, transfemoral guiding catheter receiver coil (GCC), MRI-compatible needle, and tableside consoles. Commercial real-time imaging software was customized to facilitate catheter navigation and visualization of injections at 4 completely refreshed frames per second. The aorta was traversed and the left ventricular cavity was entered under direct rtMRI guidance. Pigs underwent multiple injections with dilute gadolinium-DTPA. All myocardial segments were readily accessed. The active GCC and the passive Stiletto needle injector were readily visualized. More than 50 endomyocardial injections were performed with the aid of rtMRI; 81% were successful with this first-generation prototype. CONCLUSION: Percutaneous endomyocardial drug delivery is feasible with the aid of rtMRI, which permits precise 3-dimensional localization of injection within the LV wall. PMID- 11901037 TI - Randomized trial of a distal embolic protection device during percutaneous intervention of saphenous vein aorto-coronary bypass grafts. AB - BACKGROUND: Stents provide effective treatment for stenotic saphenous venous aorto-coronary bypass grafts, but their placement carries a 20% incidence of procedure-related complications, which potentially are related to the distal embolization of atherosclerotic debris. We report the first multicenter randomized trial to evaluate use of a distal embolic protection device during stenting of such lesions. METHODS AND RESULTS: Of 801 eligible patients, 406 were randomly assigned to stent placement over the shaft of the distal protection device, and 395 were assigned to stent placement over a conventional 0.014-inch angioplasty guidewire (control group). The primary end point-a composite of death, myocardial infarction, emergency bypass, or target lesion revascularization by 30 days-was observed in 65 patients (16.5%) assigned to the control group and 39 patients (9.6%) assigned to the embolic protection device (P=0.004). This 42% relative reduction in major adverse cardiac events was driven by myocardial infarction (8.6% versus 14.7%, P=0.008) and "no-reflow" phenomenon (3% versus 9%, P=0.02). Clinical benefit was seen even when platelet glycoprotein IIb/IIIa receptor blockers were administered (61% of patients), with composite end points occurring in 10.7% of protection device patients versus 19.4% of control patients (P=0.008). CONCLUSIONS: Use of this distal protection device during stenting of stenotic venous grafts was associated with a highly significant reduction in major adverse events compared with stenting over a conventional angioplasty guidewire. This demonstrates the importance of distal embolization in causing major adverse cardiac events and the value of embolic protection devices in preventing such complications. PMID- 11901038 TI - Angiogenic Gene Therapy (AGENT) trial in patients with stable angina pectoris. AB - BACKGROUND: The angiogenic response to myocardial ischemia can be augmented in animal models by gene transfer with the use of a replication defective adenovirus (Ad) containing a human fibroblast growth factor (FGF) gene. METHODS AND RESULTS: The objectives of the Angiogenic GENe Therapy (AGENT) trial were to evaluate the safety and anti-ischemic effects of 5 ascending doses of Ad5-FGF4 in patients with angina and to select potentially safe and effective doses for subsequent study. Seventy-nine patients with chronic stable angina Canadian Cardiovascular Society class 2 or 3 underwent double-blind randomization (1:3) to placebo (n=19) or Ad5-FGF4 (n=60). Safety evaluations were performed at each visit and exercise treadmill testing (ETT) at baseline and at 4 and 12 weeks. Single intracoronary administration of Ad5-FGF4 seemed to be safe and well tolerated with no immediate adverse events. Fever of <1-day duration occurred in 3 patients in the highest dose group. Transient, asymptomatic elevations in liver enzymes occurred in 2 patients in lower-dose groups. Serious adverse events during follow-up (mean, 311 days) were not different between placebo and Ad5-FGF4. Overall, patients who received Ad5-FGF4 tended to have greater improvements in exercise time at 4 weeks (1.3 versus 0.7 minutes, P=NS, n=79). A protocol-specified, subgroup analysis showed the greatest improvement in patients with baseline ETT < or =10 minutes (1.6 versus 0.6 minutes, P=0.01, n=50). CONCLUSIONS: Results show evidence of favorable anti-ischemic effects with Ad5-FGF4 compared with placebo, and it appears to be safe. Angiogenic gene transfer with Ad5-FGF4 shows promise as a new therapeutic approach to the treatment of angina pectoris. PMID- 11901039 TI - Effect of azithromycin treatment on endothelial function in patients with coronary artery disease and evidence of Chlamydia pneumoniae infection. AB - BACKGROUND: It has been suggested that infection with Chlamydia pneumoniae(CPn) can trigger inflammatory mechanisms that may in turn impair vascular endothelial function. The aim of the present study was to assess whether treatment with the macrolide antibiotic azithromycin improves endothelial function in patients with coronary artery disease and antibodies positive to CPn. METHODS AND RESULTS: We carried out a randomized, prospective, double-blind, placebo-controlled trial in 40 male patients (mean age, 55+/-9 years) with documented coronary artery disease and positive CPn-IgG antibody titers. After baseline evaluation, patients were randomized to receive either azithromycin or placebo for 5 weeks. Flow-mediated dilation (FMD) of the brachial artery and E-selectin, von Willebrand factor, and C-reactive protein (CRP) levels were assessed at study entry and at the end of the treatment period. Our results showed that patients who received azithromycin had a significant improvement in FMD (mean change, 2.1+/-1.1%; P<0.005). In contrast, FMD was not significantly changed in the placebo group (mean change, 0.02+/-0.2%, P=0.64). Azithromycin therapy also resulted in a significant decrease of E-selectin and von Willebrand factor levels. CRP levels were not significantly altered by treatment with either azithromycin or placebo. Beneficial effects of azithromycin treatment were independent from the presence of low (<1:32) or high (> or =1:32) CPn antibody titers. CONCLUSIONS: Our findings indicate that treatment with azithromycin has a favorable effect on endothelial function in patients with documented coronary artery disease and evidence of CPn infection irrespective of antibody titer levels. Whether these favorable actions of antibiotic treatment will translate into a beneficial effect on atherogenesis and cardiac events needs further investigation. PMID- 11901040 TI - Effects of long-term biventricular stimulation for resynchronization on echocardiographic measures of remodeling. AB - BACKGROUND: Long-term ventricular resynchronization therapy improves symptom status. Changes in left ventricular remodeling have not been adequately evaluated. METHODS AND RESULTS: Fifty-three patients with systolic heart failure and bundle-branch block underwent implantation of biventricular stimulation (BVS) devices as part of a randomized trial. Echocardiograms were acquired at randomization and at 6-week intervals until completion of 12 weeks of continuous BVS. There were no changes in heart rate or QRS duration after 12 weeks of BVS. Serum norepinephrine values did not change with BVS. After 12 weeks of BVS, left atrial volume decreased. Left ventricular end-systolic and end-diastolic dimensions and left ventricular end-systolic volume also decreased after 12 weeks of BVS. Sphericity index did not change. Measures of systolic function, including left ventricular outflow tract and aortic velocity time integral and myocardial performance index, improved. CONCLUSIONS: Long-term resynchronization therapy results in atrial and ventricular reverse remodeling and improved hemodynamics. PMID- 11901041 TI - Sex differences in the association between proinsulin and intact insulin with coronary heart disease in nondiabetic older adults: the Rancho Bernardo Study. AB - BACKGROUND: Insulin or insulin resistance is considered a coronary heart disease (CHD) risk factor, but proinsulin may have a stronger association with CHD than insulin. The role of sex differences in this association is unclear. We examined the cross-sectional association of proinsulin and insulin with CHD in older men and women without diabetes. METHODS AND RESULTS: A cross-sectional study of community-dwelling men (n=554) and women (n=902), 50 to 97 years of age, without diabetes by history or oral glucose tolerance test, was done between 1992 and 1996; plasma levels of intact insulin, proinsulin, and C-peptide were measured by radioimmunoassay. Based on questionnaire, medical history, or ECG abnormalities, 25% of men (n=136) and 24% of women (n=214) had prevalent CHD. All insulin variables were positively correlated with CHD risk factors. Compared with those without CHD, men and women with CHD had significantly higher levels of proinsulin. Women but not men with CHD also had higher levels of C-peptide and fasting and postchallenge insulin. Only proinsulin was significantly and independently associated with prevalent CHD in both men (OR=2.41, 1.42 to 4.11) and women (OR=1.80, 1.22 to 2.64) (adjusted for age, body mass index, systolic blood pressure, and HDL cholesterol). Similar analyses for fasting and postchallenge intact insulin and for C-peptide showed that among these three variables, only postchallenge insulin was significantly associated with CHD, and only in women. CONCLUSIONS: In older nondiabetic men and women, proinsulin was more strongly and consistently associated with CHD than was intact insulin. PMID- 11901042 TI - Left ventricular volume reduction by radiofrequency heating of chronic myocardial infarction in patients with congestive heart failure. AB - BACKGROUND: Myocardial infarct expansion and left ventricular (LV) remodeling are integral components in the evolution of chronic heart failure and predict morbidity and mortality. Radiofrequency (RF) heating and patch placement of chronic LV aneurysms caused a sustained reduction in LV infarct area and volume in an ovine infarct model. This study evaluated the effect of RF heating and epicardial patch as an adjunct to coronary artery bypass graft on LV volumes in patients with prior myocardial infarction, evidence of akinetic/dyskinetic scar, and LV ejection fraction < or =40%. METHODS AND RESULTS: Ten patients (3 female; mean age, 64+/-11 years) scheduled for coronary artery bypass graft were enrolled (Canadian Cardiovascular Society angina class 2.1+/-1.1; New York Heart Association class 3.1+/-0.5). Intraoperative digital photography demonstrated an acute 39% reduction in infarct area (n=5; P=0.01), and transesophageal ECGs demonstrated a 16% acute reduction in LV end-diastolic volumes (n=9; P=0.002) after RF treatment. There were no intraoperative or procedure-related postoperative complications, and during an average follow-up of >180 days, there have been no safety issues. All patients had complete relief of their angina and improvement in exercise tolerance. Serial transthoracic ECGs over the 6 months of follow-up after RF treatment demonstrated persistent reductions in LV end diastolic volume (29%; P<0.0001) and LV end-systolic volume (37%; P<0.0001) with improved ejection fraction (P<0.02). CONCLUSIONS: RF heating and patch placement in these 10 patients resulted in acute reduction in infarct area and ventricular volumes that were maintained 180 days after procedure. This technique may reduce the incidence of congestive heart failure and mortality in these patients and warrants investigation in larger clinical trials. PMID- 11901043 TI - Barium reduces resting blood flow and inhibits potassium-induced vasodilation in the human forearm. AB - BACKGROUND: Increasing extracellular K+ concentration within and just above the physiological range hyperpolarizes and relaxes vascular smooth muscle in vitro. These actions involve inwardly rectifying potassium channels (K(IR)) and Na+/K+ ATPase, which are inhibited, respectively, by Ba2+ and ouabain. The role (if any) of K(IR) in controlling human resistance vessel tone is unknown, and we investigated this in the forearm. METHODS AND RESULTS: Blood flow was measured by plethysmography in healthy men. Drugs and electrolytes were infused through the brachial artery. BaCl2 (4 micromol/min, also used in subsequent experiments) increased Ba2+ plasma concentration in the infused forearm to 50+/-0.8 micromol/L (mean+/-SEM) and reduced blood flow by 24+/-4% (n=8, P<0.001) without causing systemic effects. Ouabain (2.7 nmol/min), alone and with BaCl2, reduced flow by 10+/-2% and 28+/-3%, respectively (n=10). Incremental infusions of KCl (0.05, 0.1, and 0.2 mmol/min) increased flow from baseline by 1.0+/-0.2, 2.0+/-0.4, and 4.2+/-0.5 mL/min per deciliter forearm, respectively. Responses to KCl (0.2 mmol/min) were inhibited by BaCl2, alone and plus ouabain, by 60+/-9% and 88+/ 6%, respectively (both P< or =0.01). In control experiments, norepinephrine (240 pmol/min) reduced blood flow by 24+/-2% but had no significant effect on K+ induced vasodilation. BaCl2, alone or with ouabain, did not significantly influence responses to verapamil or nitroprusside. CONCLUSIONS: Ba2+ increases forearm vascular resistance. K+-induced vasodilation is selectively inhibited by Ba2+ and almost abolished by Ba2+ plus ouabain, suggesting a role for K(IR) and Na+/K+ ATPase in controlling basal tone and in K+-induced vasorelaxation in human forearm resistance vessels. PMID- 11901044 TI - Long-term effects of carotid sinus denervation on arterial blood pressure in humans. AB - BACKGROUND: After experimental carotid sinus denervation in animals, blood pressure (BP) level and variability increase markedly but normalize to preoperative levels within 10 to 14 days. We investigated the course of arterial BP level and variability after bilateral denervation of the carotid sinus baroreceptors in humans. METHODS AND RESULTS: We studied 4 women (age 41 to 63 years) who were referred for evaluation of arterial baroreflex function because of clinical suspicion of carotid sinus denervation attributable to bilateral carotid body tumor resection. The course of BP level and variability was assessed from repeated office and 24-hour ambulatory measurements (Spacelabs/Portapres) during 1 to 10 years of (retrospective) follow-up. Rapid cardiovascular reflex adjustments to active standing and Valsalva's maneuver were assessed. Office BP level increased from 132/86 mm Hg (range, 118 to 148/80 to 92 mm Hg) before bilateral surgery to 160/105 mm Hg (range, 143 to 194/90 to 116 mm Hg) 1 to 10 years after surgery. During continuous 24-hour noninvasive BP recording (Portapres), a marked BP variability was apparent in all 4 patients. Initial symptomatic hypotension on change to the upright posture and abnormal responses to Valsalva's maneuver were observed. CONCLUSIONS: Acute carotid sinus denervation, as a result of bilateral carotid body tumor resection, has a long term effect on the level, variability, and rapid reflex control of arterial BP. Therefore, in contrast to earlier experimental observations, the compensatory ability of the baroreceptor areas outside the carotid sinus seems to be of limited importance in the regulation of BP in humans. PMID- 11901045 TI - Long-term survival of dialysis patients in the United States with prosthetic heart valves: should ACC/AHA practice guidelines on valve selection be modified? AB - BACKGROUND: Minimal data exist on the long-term survival of dialysis patients after cardiac valve surgery. Current practice guidelines of the American College of Cardiology/American Heart Association Task Force on the management of patients with valvular heart disease proscribe the use of bioprosthetic (tissue) valves in hemodialysis patients. METHODS AND RESULTS: Dialysis patients hospitalized for heart valve replacement surgery from 1978 to 1998 were retrospectively identified from the US Renal Data System database. Long-term survival was estimated by the life-table method. The impact of demographic differences and comorbidity on outcome were examined in a Cox proportional hazards model. The in-hospital mortality of 5858 dialysis patients undergoing valve surgery was 20.7%. Aortic valve replacement was performed in 3415 patients (58%), mitral valve replacement in 1848 patients (32%), and combined aortic and mitral valve replacement in 562 patients (10%). Tissue valves were used in 881 patients. There was no significant difference in survival related to type of prosthetic valve. The 2-year survival rate was 39.7+/-3.5% with tissue valves versus 39.7+/-1.4% for nontissue valves. Compared with nontissue prosthetic valves, the use of tissue valves was not predictive of death (RR 0.98; 95% CI 0.90 to 1.07). CONCLUSIONS: There is no significant difference in survival of dialysis patients after cardiac valve replacement with tissue versus nontissue prosthetic valves. Current practice guidelines proscribing the use of bioprosthetic heart valves in hemodialysis patients should be rescinded. PMID- 11901046 TI - Natural history of Brugada syndrome: insights for risk stratification and management. AB - BACKGROUND: Treatment of patients with Brugada syndrome is complicated by the incomplete information on the natural history of the disease related to the small number of cases reported. Furthermore, the value of programmed electrical stimulation (PES) for risk stratification is highly debated. The objective of this study was to search for novel parameters to identify patients at risk of sudden death. METHODS AND RESULTS: Clinical data were collected for 200 patients (152 men, 48 women; age, 41+/-18 years) and stored in a dedicated database. Genetic analysis was performed, and mutations on the SCN5A gene were identified in 28 of 130 probands and in 56 of 121 family members. The life-table method of Kaplan-Meier used to define the cardiac arrest-free interval in patients undergoing PES failed to demonstrate an association between PES inducibility and spontaneous occurrence of ventricular fibrillation. Multivariate Cox regression analysis showed that after adjusting for sex, family history of sudden death, and SCN5A mutations, the combined presence of a spontaneous ST-segment elevation in leads V1 through V3 and the history of syncope identifies subjects at risk of cardiac arrest (HR, 6.4; 95% CI, 1.9 to 21; P<0.002). CONCLUSIONS: The information on the natural history of patients obtained in this study allowed elaboration of a risk-stratification scheme to quantify the risk for sudden cardiac death and to target the use of the implantable cardioverter defibrillator. PMID- 11901047 TI - BG9719 (CVT-124), an A1 adenosine receptor antagonist, protects against the decline in renal function observed with diuretic therapy. AB - BACKGROUND: Adenosine may adversely affect renal function via its effects on renal arterioles and tubuloglomerular feedback, but effects of adenosine blockade in humans receiving furosemide and ACE inhibitors is unknown. METHODS AND RESULTS: This was a randomized, double-blind, ascending-dose, crossover study evaluating 3 doses of BG9719 in 63 patients with congestive heart failure. Patients received placebo or 1 of 3 doses of BG9719 on 1 day and the same medication plus furosemide on a separate day. Renal function and electrolyte and water excretion were assessed. BG9719 alone caused an increase in urine output and sodium excretion (P<0.05). Although administration of furosemide alone caused a large diuresis, addition of BG9719 to furosemide increased diuresis, which was significant at the 0.75-microg/mL concentration. BG9719 alone improved glomerular filtration rate (GFR) at the 2 lower doses. Furosemide alone caused a decline in GFR. When BG9719 was added to furosemide, however, creatinine clearance remained at baseline at the 2 lower doses. CONCLUSIONS: In patients with congestive heart failure on standard therapy, including ACE inhibitors, BG9719 increased both urine output and GFR. In these same patients, furosemide increased urine output at the expense of decreased GFR. When BG9719 was given in addition to furosemide, urine volume additionally increased and there was no deterioration in GFR. A1 adenosine antagonism might preserve renal function while simultaneously promoting natriuresis during treatment for heart failure. PMID- 11901048 TI - Plasma norepinephrine predicts survival and incident cardiovascular events in patients with end-stage renal disease. AB - BACKGROUND: Sympathetic tone is consistently raised in patients with end-stage renal disease (ESRD). We therefore tested the hypothesis that sympathetic activation is associated with mortality and cardiovascular events in a cohort of 228 patients undergoing chronic hemodialysis who did not have congestive heart failure at baseline and who had left ventricular ejection fraction >35%. METHODS AND RESULTS: The plasma concentration of norepinephrine (NE) was used as a measure of sympathetic activity. Plasma NE exceeded the upper limit of the normal range (cutoff 3.54 nmol/L) in 102 dialysis patients (45%). In a multivariate Cox regression model that included all univariate predictors of death as well as the use of sympathicoplegic agents and beta-blockers, plasma NE proved to be an independent predictor of this outcome (hazard ratio [1-nmol/L increase in plasma NE]: 1.07, 95% CI 1.01 to 1.14, P=0.03). Similarly, plasma NE emerged as an independent predictor of fatal and nonfatal cardiovascular events (hazard ratio [1-nmol/L increase in plasma NE] 1.08, 95% CI 1.02 to 1.15, P=0.01) in a model that included previous cardiovascular events, pulse pressure, age, diabetes, smoking, and use of sympathicoplegic agents and beta-blockers. The adjusted relative risk for cardiovascular complications in patients with plasma NE >75th percentile was 1.92 (95% CI 1.20 to 3.07) times higher than in those below this threshold (P=0.006). CONCLUSIONS: Sympathetic nerve overactivity is associated with mortality and cardiovascular outcomes in ESRD. Controlled trials with antiadrenergic drugs are needed to determine whether interference with the sympathetic system could reduce the high cardiovascular morbidity and mortality in dialysis patients. PMID- 11901049 TI - Maternal hypercholesterolemia during pregnancy promotes early atherogenesis in LDL receptor-deficient mice and alters aortic gene expression determined by microarray. AB - BACKGROUND: Maternal hypercholesterolemia during pregnancy is associated with markedly enhanced fatty streak formation in human fetal aortas and accelerated progression of atherosclerosis in normocholesterolemic children. METHODS AND RESULTS: To establish the causal role of maternal hypercholesterolemia in a genetically homogeneous murine model and to test the hypothesis that pathogenic events during fetal development result in persistent changes in arterial gene expression, female LDL receptor-deficient (LDLR(-/-)) mice were fed regular chow or high-fat diets supplemented with 0.075% or 1.25% cholesterol during pregnancy. Lesion sizes were determined in the aortic origin of their chow-fed offspring at 3 months. Maternal hypercholesterolemia more than doubled lesion sizes in male offspring (P<0.0001 for the 0.0075% cholesterol group). Microarray analysis of the expression of 11 000 murine genes in the nonatherosclerotic descending aorta by Affymetrix gene chips suggested that 139 genes were significantly regulated in offspring of hypercholesterolemic mothers. A subset of 12 of the upregulated transcripts was subjected to secondary analysis by semiquantitative PCR of pooled RNA and 4 genes were found to be upregulated >1.7-fold. Quantitative PCR for one of these genes using RNA from individual mice yielded similar results. Comparative immunostaining for several of the above genes also indicated increased protein content in offspring of hypercholesterolemic mothers. CONCLUSIONS: These findings establish an atherogenic effect of maternal hypercholesterolemia in genetically uniform mice and indicate that changes in aortic gene expression persist long after fetal exposure to hypercholesterolemia. In addition to elucidating pathogenic mechanisms initiated during fetal development, this approach may identify genes in morphologically normal arteries that influence the susceptibility to classical risk factors of atherosclerosis. PMID- 11901050 TI - Nongenomic mechanisms of endothelial nitric oxide synthase activation by the selective estrogen receptor modulator raloxifene. AB - BACKGROUND: Nontranscriptional signaling through estrogen receptors (ERs) is important in the cardiovascular system. In particular, estrogen stimulates endothelial NO synthase (eNOS) via the phosphatidylinositol 3-kinase (PI3K) pathway. The selective estrogen receptor modulator (SERM) raloxifene is effective for the treatment of postmenopausal osteoporosis, but its ability to activate eNOS via PI3K is unknown. METHODS AND RESULTS: Human umbilical vein endothelial cells were cultured in estrogen-deprived, phenol red-free medium. Raloxifene stimulated eNOS in a concentration- and time-dependent manner. Activation of eNOS by raloxifene was blocked by the PI3K inhibitor wortmannin and by the ER antagonist ICI 182,780 but not by transcriptional or translational inhibitors. Coimmunoprecipitation studies demonstrated that, in a ligand-dependent manner, raloxifene increased ERalpha-associated p85alpha, p110alpha, and PI3K activity. This correlated temporally with increases in the serine and threonine phosphorylation and activation of protein kinase Akt. CONCLUSIONS: Our findings indicate that nongenomic ER signaling triggered by a SERM leads to a rapid activation of NO synthesis in human endothelial cells. The ability of raloxifene to facilitate ERalpha-PI3K interaction may provide additional insight into the structure-function relationship of specific SERMs, which promote the nontranscriptional effects of ER. PMID- 11901051 TI - Propranolol prevents the development of heart failure by restoring FKBP12.6 mediated stabilization of ryanodine receptor. AB - BACKGROUND: In heart failure, protein kinase A-mediated hyperphosphorylation of ryanodine receptors (RyRs) in sarcoplasmic reticulum (SR) causes dissociation of FKBP12.6 from RyRs. This results in an abnormal Ca2+ leak through RyRs, possibly leading to cardiac dysfunction. In the present study, we assess whether beta blockers can correct this defect in RyR in tachycardia-induced heart failure and thereby improve cardiac function. METHODS AND RESULTS: SRs were isolated from dog left ventricular muscles (normal group, 4 weeks of rapid right ventricular pacing with or without propranolol [P(+) or P(-)]). End-diastolic and end-systolic diameters both increased less in P(+) than P(-), associated with a smaller decrease in fractional shortening in P(+). In SR from P(-), a prominent Ca2+ leak was observed, and FK506 (which dissociates FKBP12.6 from RyR) did not induce an additional Ca2+ leak. However, there was no appreciable Ca2+ leak in SR from P(+), although FK506 induced a Ca2+ leak as in normal SRs. In SR from P(+), an FK506-induced conformational change in RyR, which was virtually absent in SR from P(-), was observed as in normal SRs. Both the stoichiometry of FKBP12.6 versus RyR, assessed by [3H]FK506 and [3H]ryanodine binding assays, and the protein expression of FKBP12.6, assessed by Western blot analysis, were restored by propranolol toward the levels seen in normal SRs. CONCLUSIONS: Low-dose propranolol corrects the defective interaction of FKBP12.6 with RyR (restoration of RyR conformational change and prevention of Ca2+ leak from RyR), apparently resulting in an attenuation of intracellular Ca2+ overload and hence preventing the development of left ventricular remodeling in heart failure. PMID- 11901053 TI - New concepts in diastolic dysfunction and diastolic heart failure: Part I: diagnosis, prognosis, and measurements of diastolic function. PMID- 11901052 TI - Effects of cariporide and losartan on hypertrophy, calcium transients, contractility, and gene expression in congestive heart failure. AB - BACKGROUND: The purpose of this study was to compare long-term effects of cariporide with those of losartan in postinfarction heart failure. METHODS AND RESULTS: Female Sprague-Dawley rats with large myocardial infarctions and sham controls were randomized to losartan, cariporide, or placebo after 7 days and treated for 49 days. Cardiac function was assessed by echocardiography and measurement of left ventricular pressures, and gene expression was assessed by competitive reverse transcription-polymerase chain reaction. Cell dimensions, shortening, and relaxation were determined by videomicroscopy and calcium transients by fura 2. Losartan reduced postinfarction systolic and diastolic left ventricular dilation (by 24% and 31%, respectively), left and right ventricular weight (by 22% and 26%, respectively), and cardiomyocyte hypertrophy length and width (by 62% and 54%, respectively). Induction of myocardial atrial natriuretic peptide decreased 66%. Cariporide did not affect postinfarction hypertrophy or atrial natriuretic peptide. Losartan and cariporide respectively improved reduced cellular contractility (55% and 30%) and reduced elevated systolic (86% and 27%) and diastolic (49% and 43%) calcium. Losartan and cariporide respectively reduced prolonged time to 50% relaxation (66% and 25%) and time to 50% calcium reduction (55% and 53%). CONCLUSIONS: Losartan and cariporide improve cardiomyocyte contractility and calcium regulation in chronic heart failure. Losartan has salutary effects on postinfarction remodeling and gene expression, whereas cariporide is neutral. PMID- 11901054 TI - Images in cardiovascular medicine. Left ventricular involvement in arrhythmogenic right ventricular cardiomyopathy. PMID- 11901055 TI - Images in cardiovascular medicine. Premature closure of the foramen ovale and ductus arteriosus in a fetus with transposition of the great arteries. PMID- 11901056 TI - Angiogenic gene therapy appears safe to treat stable angina. PMID- 11901057 TI - Body iron stores and coronary heart disease. PMID- 11901058 TI - Erythrocyte galactose 1-phosphate quantified by isotope-dilution gas chromatography-mass spectrometry. AB - BACKGROUND: Measurements of alpha-D-galactose 1-phosphate (Gal-1-P) in erythrocytes are used to monitor the adequacy of dietary therapy in the treatment of galactosemia. We have devised a gas chromatography-mass spectrometry (GC/MS) isotope-dilution method for quantification of Gal-1-P. METHODS: We prepared trimethylsilyl (TMS) derivatives and used alpha-D-[2-(13)C]Gal-1-P as the internal standard for GC/MS. Results obtained with this method were compared with those determined by the established enzymatic method for samples from 23 healthy individuals (11 children and 12 adults), 9 suspected patients with galactosemia, 12 galactosemic patients on diet therapy, and 2 newly diagnosed toxic neonates. RESULTS: The method was linear up to 2.5 mmol/L with a lower limit of detection of 2.1 nmol (0.55 mg/L). Intra- and interassay imprecision (CVs) was 2.2-8.8%. In the 23 healthy individuals, values ranged from nondetectable to 9.2 micromol/L (2.4 mg/L of packed erythrocytes). Galactosemic patients on diet therapy had values of 10.9-45 mg/L of packed erythrocytes, whereas the newly identified patients had values of 166 and 373 mg/L. CONCLUSIONS: The GC/MS method is precise and useful over the wide range of concentrations needed to assess the galactose burden in patients with galactosemia. PMID- 11901059 TI - Immunometric assay interference: incidence and prevention. AB - BACKGROUND: The primary aim of the study was to reduce interference in an in house two-site, two-step immunometric assay. METHODS: In the running laboratory routine, 11 261 samples were tested with a carcinoembryonic antigen (CEA) assay with bovine immunoglobulin but no murine immunoglobulins in the buffer, in parallel to our routine CEA assay, using 15 mg/L heat-treated nonspecific murine immunoglobulin (MAK33) in the buffer and with the Fc fragments removed from the capture antibody. RESULTS: The frequency of interference was estimated to be 4.0% (95% confidence interval, 3.3-4.7%). The addition of 15 mg/L native MAK33 had little effect (frequency, 3.9%; 95% confidence interval, 3.2-4.6%), whereas adding 15 mg/L heat-treated MAK33 reduced interference to 0.86% (0.61-1.12%), and adding 50 mg/L reduced it further to 0.06% (0-0.13%). Removing the Fc fragments by itself reduced interference to 0.10% (0.02-0.19%). There were no statistically significant differences for age (P <0.23) or gender (P <0.40) between patients with interference (n = 210) and a randomly selected interference-negative control group (n = 186). Interference was not constant in patients: 15 of 25 individuals positive for interference and with four or more samples screened for interference had an interference-negative sample either before or after the peak of interference. CONCLUSIONS: In a two-site, two-step immunometric assay using mouse monoclonal antibodies, use of heat-treated nonspecific murine immunoglobulin in the buffer or removal of the Fc fragment from the capture antibody could improve performance. PMID- 11901060 TI - Biochemical and genetic markers of iron status and the risk of coronary artery disease: an angiography-based study. AB - BACKGROUND: Iron may promote coronary atherosclerotic disease (CAD) by increasing lipid peroxidation. Studies on biochemical or genetic markers of body iron stores as risk factors for CAD have yielded conflicting results. METHODS: We studied 849 individuals with a clear-cut definition of the CAD phenotype, i.e., with (CAD; n = 546) or without (CAD-free; n = 303) angiographically documented disease. We determined serum ferritin, as a biochemical estimate of iron stores, and the C282Y mutation in the HFE gene, i.e., the main cause of hemochromatosis in Caucasians. The relationships of ferritin with serum markers of either inflammation [C-reactive protein (CRP)] or lipid peroxidation (malondialdehyde) were also investigated. RESULTS: Mean ferritin concentrations were slightly higher in CAD vs CAD-free individuals, but this difference disappeared after adjusting for sex and CRP. Ferritin was significantly correlated with CRP (Spearman's test, rho = 0.129; P <0.001). Heterozygotes for Cys282Tyr were 4.8% among the CAD group and 6.6% among the CAD-free group (P = 0.26). The prevalence of high concentrations of stored iron, defined as ferritin concentrations above the sex-specific upper quintiles of the control distribution, was also similar in the two groups. There was a higher prevalence of "iron depletion" in CAD-free vs CAD females (20% vs 8.8%, respectively), but this difference disappeared after adjustment for age and other cardiovascular risk factors (odds ratio, 0.66; 95% confidence interval, 0.21-2.08). No differences in iron markers were found in CAD patients with or without myocardial infarction. CONCLUSIONS: Our results do not support a role for biochemical or genetic markers of iron stores as predictors of the risk of CAD or its thrombotic complications. PMID- 11901061 TI - Detection of epitestosterone doping by isotope ratio mass spectrometry. AB - BACKGROUND: Epitestosterone is prohibited by sport authorities because its administration will lower the urinary testosterone/epitestosterone ratio, a marker of testosterone administration. A definitive method for detecting epitestosterone administration is needed. METHODS: We developed a gas chromatography-combustion-isotope ratio mass spectrometry method for measuring the delta(13)C values for urinary epitestosterone. Sample preparation included deconjugation with beta-glucuronidase, solid-phase extraction, and semipreparative HPLC. Epitestosterone concentrations were determined by gas chromatography-mass spectrometry for urines obtained from a control group of 456 healthy males. Epitestosterone delta(13)C values were determined for 43 control urines with epitestosterone concentrations > or =40 microg/L (139 nmol/L) and 10 athletes' urines with epitestosterone concentrations > or =180 microg/L (624 nmol/L), respectively. RESULTS: The log epitestosterone concentration distribution was gaussian [mean, 3.30; SD, 0.706; geometric mean, 27.0 microg/L (93.6 nmol/L)]. The delta(13)C values for four synthetic epitestosterones were low (less than or equal to -30.3 per thousand) and differed significantly (P <0.0001). The SDs of between-assay precision studies were low (< or =0.73 per thousand). The mean delta(13)C values for urine samples obtained from 43 healthy males was -23.8 per thousand (SD, 0.93 per thousand). Nine of 10 athletes' urine samples with epitestosterone concentrations >180 microg/L (624 nmol/L) had delta(13)C values within +/- 3 SD of the control group. The delta(13)C value of epitestosterone in one sample was -32.6 per thousand (z-score, 9.4), suggesting that epitestosterone was administered. In addition, the likelihood of simultaneous testosterone administration was supported by low delta(13)C values for androsterone and etiocholanolone. CONCLUSIONS: Determining delta(13)C values for urinary epitestosterone is useful for detecting cases of epitestosterone administration because the mean delta(13)C values for a control group is high ( 23.8 per thousand) compared with the delta(13)C values for synthetic epitestosterones. PMID- 11901062 TI - Candidate reference method for total thyroxine in human serum: use of isotope dilution liquid chromatography-mass spectrometry with electrospray ionization. AB - BACKGROUND: There is a need for a critically evaluated reference method for thyroxine to provide an accuracy base to which routine methods can be traceable. We describe a candidate reference method involving isotope-dilution coupled with liquid chromatography/mass spectrometry. METHODS: An isotopically labeled internal standard, thyroxine-d(5), was added to serum, followed by equilibration, protein precipitation, and ethyl acetate and solid-phase extractions to prepare samples for liquid chromatography-mass spectrometry electrospray ionization (LC/MS-ESI) analysis. For separation, a Zorbax Eclipse XDB-C(18) column was used with a mobile phase consisting of 1 mL/L acetic acid in acetonitrile-water (32:68 by volume) for positive ions and a Zorbax Extend-C(18) column with a mobile phase consisting of 2 mL/L ammonium hydroxide in methanol-water (32:68 by volume) for negative ions. [M + H](+) ions at m/z 778 and 783 for thyroxine and its labeled internal standard were monitored for positive ions and [M - H](-) ions at m/z 776 and 781 for negative ions. Samples of frozen serum pools were prepared and measured in three separate sets. RESULTS: Within-set CVs were 0.2-1.0%. The correlation coefficients of all linear regression lines (measured intensity ratios vs mass ratios) were 0.999-1.000. Positive- and negative-ion measurements agreed with a mean difference of 0.45% at three concentrations (50, 110, and 168 microg/L). The detection limits (at a signal-to-noise ratio of approximately 3 to 5) were 30 and 20 pg for positive and negative ions, respectively. The results from the LC/MS-ESI method were within 1 SD of the composite means from many routine clinical methods, although it appears that the clinical method means may be biased high by 4-5 microg/L across the concentrations. Some routine clinical methods may be biased by up to 20% at low concentrations. CONCLUSIONS: This well characterized LC/MS-ESI method for total serum thyroxine with a theoretically sound approach, demonstrated good accuracy and precision, and low susceptibility to interferences qualifies as a candidate reference method. Use of this reference method as an accuracy base may reduce the apparent biases in routine methods along with the high interlaboratory imprecision. PMID- 11901063 TI - Factors influencing serum neopterin concentrations in a population of blood donors. PMID- 11901064 TI - Serologic diagnosis of Hantaan virus infection based on a peptide antigen. PMID- 11901065 TI - Maternal nitric oxide supplementation decreases cord blood S100B in intrauterine growth-retarded fetuses. PMID- 11901066 TI - Hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome as a complication of preeclampsia in pregnant women increases the amount of cell-free fetal and maternal DNA in maternal plasma and serum. PMID- 11901067 TI - Accuracy of expected risk of Down syndrome using the second-trimester triple test. PMID- 11901068 TI - Correlation of serum immunoglobulin free light chain quantification with urinary Bence Jones protein in light chain myeloma. PMID- 11901069 TI - Genotyping of the apolipoprotein B R3500Q mutation using immobilized locked nucleic acid capture probes. PMID- 11901070 TI - Proton nuclear magnetic resonance spectral profiles of urine from children and adolescents with type 1 diabetes. PMID- 11901071 TI - Nanotechnology and applications: an all-language literature survey including books and patents. PMID- 11901072 TI - Effects of anticoagulant and time of plasma separation on measurement of homocysteine. PMID- 11901073 TI - Detection of cardiac troponin I early after onset of chest pain in six patients. PMID- 11901074 TI - Serum CrossLaps: pediatric reference intervals from birth to 19 years of age. PMID- 11901075 TI - Release kinetics of cardiac troponin T in survivors of confirmed severe pulmonary embolism. PMID- 11901076 TI - False-positive phencyclidine immunoassay results caused by venlafaxine and O desmethylvenlafaxine. PMID- 11901077 TI - False-positive troponin I attributed to a macrocomplex. PMID- 11901078 TI - Automated assay for fetal DNA analysis in maternal serum. PMID- 11901079 TI - Method-dependent changes in "HDL-cholesterol" with recombinant apolipoprotein A I(Milano) infusion in healthy volunteers. PMID- 11901086 TI - Atypical kinetic profiles in drug metabolism reactions. PMID- 11901087 TI - Polymorphisms in the ABCC2 (cMOAT/MRP2) gene found in 72 established cell lines derived from Japanese individuals: an association between single nucleotide polymorphisms in the 5'-untranslated region and exon 28. AB - We found nucleotide variability in the 5'-upstream region and exonic sequences of a gene-encoding canalicular multispecific organic anion transporter/multidrug resistance-associated protein 2 (cMOAT/MRP2) by polymerase chain reaction-based sequencing using genomic DNA from 72 established cell lines derived from 72 Japanese individuals. Four single nucleotide polymorphisms (SNPs) were found in the 5'-untranslational region and 21 in the exonic regions. Of them, 14 were nonsynonymous SNPs. One deletion of seven consecutive adenines resulting in a frameshift variant was also found. Four SNPs, c-24t, g1249a (V417I), c2366t (S789F), and c3972t (I1324I), were the same as those recently reported. A strong association was found between c-24t (5'-untranslated region) and c3972t (exon 28), with the promoter activity of the former worth being compared. PMID- 11901088 TI - The anthelminthic agent albendazole does not interact with p-glycoprotein. AB - Albendazole is a clinically important anthelminthic agent known to have variable and low oral bioavailability. The aim of this work was to determine whether albendazole, a CYP3A4 substrate, is also a substrate for the multidrug efflux transporter P-glycoprotein. Both in vitro and in vivo methods were used to assess the role of P-glycoprotein-mediated albendazole transport. In cultured LLC-PK1, L MDR1, and Caco-2 cells, albendazole was found not to be a P-glycoprotein substrate; the transport across LLC-PK1 and L-MDR1 cells revealed basal to apical versus apical to basal transport to a similar extent. In addition, there was no inhibitory effect of albendazole on digoxin transport in Caco-2 cells, and P glycoprotein inhibitors (verapamil and quinidine) did not affect transport across Caco-2 cells. The in vivo relevance of P-glycoprotein to albendazole disposition was assessed using mdr1a/1b(-/-) mice after intravenous administration of albendazole (15 mg/kg). A similar pattern of tissue distribution in both P glycoprotein-deficient and wild-type mice was observed. In conclusion, albendazole is neither a substrate nor an inhibitor of P-glycoprotein. Therefore, interactions between albendazole and P-glycoprotein substrates or inhibitors are unlikely to be clinically important. PMID- 11901089 TI - Absorption and metabolism of flavonoids in the caco-2 cell culture model and a perused rat intestinal model. AB - The purpose of present study was to determine the intestinal absorption and metabolism of genistein and its analogs to better understand the mechanisms responsible for their low oral bioavailability. The Caco-2 cell culture model and a perfused rat intestinal model were used for the study. In both models, permeabilities of aglycones (e.g., genistein) were comparable to well absorbed compounds, such as testosterone and propranolol. In the Caco-2 model, permeabilities of aglycones were at least 5 times higher (p < 0.05) than their corresponding glycosides (e.g., genistin), and the vectorial transport of aglycones was similar (p > 0.05). In contrast, vectorial transport of glucosides favored excretion (p < 0.05). Limited hydrolysis of glycosides was observed in the Caco-2 model, which was completely inhibited (p < 0.05) by 20 mM gluconolactone, a broad specificity glycosidase inhibitor. In the perfused rat intestinal model, genistin was rapidly hydrolyzed (about 40% in 15 min) in the upper intestine but was not hydrolyzed at all in the colon. Aglycones were rapidly absorbed (P*(eff) > 1.5), and absorbed aglycones underwent extensive (40% maximum) phase II metabolism via glucuronidation and sulfation in the upper small intestine. Similar to the hydrolysis, recovery of conjugated genistein was also region-dependent, with jejunum having the highest and colon the lowest (p < 0.05). This difference in conjugate recovery could be due to the difference in the activities of enzymes or efflux transporters, and the results of studies tend to suggest that both of these factors were involved. In conclusion, genistein and its analogs are well absorbed in both intestinal models, and therefore, poor absorption is not the reason for its low bioavailability. On the other hand, extensive phase II metabolism in the intestine significantly contributes to its low bioavailability. PMID- 11901090 TI - In vitro effect of standardized ginseng extracts and individual ginsenosides on the catalytic activity of human CYP1A1, CYP1A2, and CYP1B1. AB - Ginseng extract has been reported to decrease the incidence of 7,12 dimethylbenz[a]anthracene (DMBA)-initiated tumorigenesis in mice. A potential mechanism for this effect by ginseng is inhibition of DMBA-bioactivating cytochrome P450 (P450) enzymes. In the present in vitro study, we examined the effect of a standardized Panax ginseng (or Asian ginseng) extract (G115), a standardized Panax quinquefolius (or North American ginseng) extract (NAGE), and individual ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) on CYP1 catalytic activities, as assessed by 7-ethoxyresorufin O-dealkylation. G115 and NAGE decreased human recombinant CYP1A1, CYP1A2, and CYP1B1 activities in a concentration-dependent manner. Except for the competitive inhibition of CYP1A1 by G115, the mode of inhibition was the mixed-type in the other cases. A striking finding was that NAGE was 45-fold more potent than G115 in inhibiting CYP1A2. Compared with G115, NAGE also preferentially inhibited 7-ethoxyresorufin O dealkylation activity in human liver microsomes. Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1, either individually or as a mixture and at the levels reflecting those found in an inhibitory concentration (100 microg/ml) of NAGE or G115, did not influence CYP1 activities. However, at a higher ginsenoside concentration (50 microg/ml), Rb1, Rb2, Rc, Rd, and Rf inhibited these activities. Overall, our in vitro findings indicate that standardized NAGE and G115 extracts, which were not treated with calf serum or subjected to acid hydrolysis, inhibited CYP1 catalytic activity in an enzyme-selective and extract-specific manner, but the effects were not due to Rb1, Rb2, Rc, Rd, Re, Rf, or Rg1. PMID- 11901091 TI - Polymorphic variants (CYP2C9*3 and CYP2C9*5) and the F114L active site mutation of CYP2C9: effect on atypical kinetic metabolism profiles. AB - CYP2C9 wild-type protein has been shown to exhibit atypical kinetic profiles of metabolism that may affect in vitro-in vivo predictions made during the drug development process. Previous work suggests a substrate-dependent effect of polymorphic variants of CYP2C9 on the rate of metabolism; however, it is hypothesized that these active site amino acid changes will affect the kinetic profile of a drug's metabolism as well. To this end, the kinetic profiles of three model CYP2C9 substrates (flurbiprofen, naproxen, and piroxicam) were studied using purified CYP2C9*1 (wild-type) and variants involving active site amino acid changes, including the naturally occurring variants CYP2C9*3 (Leu359) and CYP2C9*5 (Glu360) and the man-made mutant CYP2C9 F114L. CYP2C9*1 (wild-type) metabolized each of the three compounds with a distinctive profile reflective of typical hyperbolic (flurbiprofen), biphasic (naproxen), and substrate inhibition (piroxicam) kinetics. CYP2C9*3 metabolism was again hyperbolic for flurbiprofen, of a linear form for naproxen (no saturation noted), and exhibited substrate inhibition with piroxicam. CYP2C9*5-mediated metabolism was hyperbolic for flurbiprofen and piroxicam but linear with respect to naproxen turnover. The F114L mutant exhibited a hyperbolic kinetic profile for flurbiprofen metabolism, a linear profile for naproxen metabolism, and a substrate inhibition kinetic profile for piroxicam metabolism. In all cases except F114L-mediated piroxicam metabolism, turnover decreased and the K(m) generally increased for each allelic variant compared with wild-type enzyme. It seems that the kinetic profile of CYP2C9-mediated metabolism is dependent on both substrate and the CYP2C9 allelic variant, thus having potential ramifications on drug disposition predictions made during the development process. PMID- 11901092 TI - Interaction of irinotecan (CPT-11) and its active metabolite 7-ethyl-10 hydroxycamptothecin (SN-38) with human cytochrome P450 enzymes. AB - The inhibition and mechanism-based inactivation potencies of irinotecan (7-ethyl 10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin; CPT-11) and its active metabolite (7-ethyl-10-hydroxycamptothecin; SN-38) for human cytochrome P450 (P450) enzymes were investigated to evaluate the potential for drug interactions involving CPT-11 using microsomes from insect cells expressing specific human P450 isoforms. The mechanism and potential for interaction were examined by Lineweaver-Burk analysis, and NADPH-, time- and concentration-dependent effects were observed. CPT-11 and SN-38 competitively inhibited CYP3A4 (testosterone 6 beta-hydroxylation) activity with K(i) values of 129 and 121 microM, respectively. CYP2A6 (coumarin 7-hydroxylation) and CYP2C9 (diclofenac 4' hydroxylation) activities exhibited a mixed type of inhibition comprising competitive and noncompetitive components in response to SN-38, the K(i) values being 181 and 156 microM, respectively. On the other hand, CYP1A2 (phenacetin O deethylation), CYP2B6 (7-ethoxycoumarin O-deethylation), CYP2C8 (paclitaxel 6 alpha-hydroxylation), CYP2C19 (S-mephenytoin 4'-hydroxylation), CYP2D6 (bufuralol 1'-hydroxylation), and CYP2E1 (chlorzoxazone 6-hydroxylation) were hardly affected by either compound. Furthermore, CPT-11 and SN-38 were suggested to be mechanism-based inactivators of CYP3A4. The k(inact) and K(I) values of CPT-11 and SN-38 were 0.06 min(-1) and 24 microM and 0.10 min(-1) and 26 microM, respectively. However, no inactivation of CYP2A6 and CYP2C9 by SN-38 was observed. These results mean that CPT-11 and SN-38 interact with human P450 isoforms, such as CYP2A6, CYP2C9, and CYP3A4, in vitro and imply that the significant drug interactions involving CPT-11 may be caused by a mechanism-based inactivation of CYP3A4 by SN-38 as an active metabolite of CPT-11 rather than competitive inhibition. PMID- 11901093 TI - Glucuronidation: an important mechanism for detoxification of benzo[a]pyrene metabolites in aerodigestive tract tissues. AB - UDP-glucuronosyltransferases (UGTs) have been implicated as important detoxifying enzymes for several major tobacco carcinogens. Because the aerodigestive tract is a primary target for exposure to tobacco smoke carcinogens, the major goal of the present study was to determine whether aerodigestive tract tissues exhibit glucuronidating activity against metabolites of benzo[a]pyrene (BaP) and to explore the pattern of expression of UGT genes in a series of aerodigestive tract tissue specimens. Glucuronidation of the phenolic BaP metabolites 3-, 7-, and 9 hydroxy-BaP was observed in all upper aerodigestive tract tissue microsome specimens tested, as determined by high-pressure liquid chromatography analysis. Glucuronidating activity toward the procarcinogenic BaP metabolite trans-BaP-7,8 dihydrodiol(+/-) was also detected in aerodigestive tract tissues. By semiquantitative duplex reverse transcription-polymerase chain reaction analysis, UGT1A7 and UGT1A10 were shown to be well expressed in all aerodigestive tract tissues examined, including tongue, tonsil, floor of mouth, larynx, and esophagus. UGT1A8 and UGT1A6 were expressed primarily in larynx; no expression was observed for UGTs 1A1, 1A3, 1A4, 1A5, 1A9. Of the family 2B UGTs, only UGT2B4 and UGT2B17 exhibited significant levels of expression in aerodigestive tract tissues. Of the aerodigestive tract-expressing UGTs, only UGTs 1A7, 1A8, and 1A10 exhibited glucuronidating activity against 7-hydroxy-BaP, with UGT1A10 exhibiting the highest affinity as determined by kinetic analysis (K(m) = 49 microM). No UGT expression or glucuronidating activity was observed for any of the lung specimens analyzed in this study. These results suggest that several family 1 UGTs may potentially play an important role in BaP detoxification in the aerodigestive tract. PMID- 11901094 TI - Development of an in vitro screening model for the biosynthesis of acyl glucuronide metabolites and the assessment of their reactivity toward human serum albumin. AB - An in vitro screening model was developed to determine the reactivity of acyl glucuronide metabolites from carboxylic drugs. This assay is composed of two phases. The first is a phase of biosynthesis of acyl glucuronides by human liver microsomes (HLM). The second, during which acyl glucuronides are incubated with human serum albumin (HSA), consists of assessing the reactivity of acyl glucuronides toward HSA. Both phases are performed successively in the same experiment. This model was validated using eight carboxylic drugs that were well known for their reactivity, their extent of covalent binding, and their immunological potential. These products were representative of the scale of reactivity. Each compound was incubated with HLM at 400 microM and metabolized into acyl glucuronide to different extents, ranging from 5.6% (tolmetin) to 89.4% (diclofenac). The first-order aglycone appearance rate constant and the extent of covalent binding to proteins were assayed during the incubation of acyl glucuronides formed with HSA for 24 h. Extensive isomerization phenomenon was observed for each acyl glucuronide between the two phases. An excellent correlation was observed (r(2), 0.94) between the extent of drug covalent binding to albumin and the aglycone appearance constant weighted by the percentage of isomerization. This correlation represents an in vitro reactivity scale, which will be helpful in drug discovery support programs to predict the covalent binding potential of new chemical entities. This screening model will also allow the comparison of acyl glucuronide reactivity for related structure compounds. PMID- 11901095 TI - Reductive metabolism of an alpha,beta-ketoalkyne, 4-phenyl-3-butyn-2-one, by rat liver preparations. AB - The reduction of the triple bond and carbonyl group of an alpha,beta-ketoalkyne, 4-phenyl-3-butyn-2-one (PBYO), by rat liver microsomes and cytosol was investigated. The triple-bond-reduced product trans-4-phenyl-3-buten-2-one (PBO) and the carbonyl-reduced product 4-phenyl-3-butyn-2-ol (PBYOL) were formed when PBYO was incubated with rat liver microsomes in the presence of NADPH. The triple bond of 1-phenyl-1-butyne, deprenyl, ethynylestradiol, ethinamate, and PBYOL, in which the triple bond is not adjacent to a carbonyl group, were not reduced by liver microsomes even in the presence of NADPH. PBO was further reduced to 4 phenyl-2-butanone (PBA) by liver cytosol with NADPH. PBYOL was also formed from PBYO by liver cytosol in the presence of NADPH or NADH. The microsomal triple bond reductase activity was inhibited by disulfiram, 7-dehydrocholesterol, and 18 beta-glycyrrhetinic acid but not beta-diethylaminoethyldiphenylpropylacetate or carbon monoxide. The triple-bond reductase activity in liver microsomes was not enhanced by several inducers of the rat cytochrome P450 system. These results suggested that the triple-bond reduction is caused by a new type of reductase, not cytochrome P450. The microsomal and cytosolic carbonyl reductase activities were not inhibited by quercitrin, indomethacin, or phenobarbital. Only S-PBYOL was formed from PBYO by liver cytosol. In contrast, liver microsomes produced R PBYOL together with the S-enantiomer to some extent. Ethoxyresorufin-O-dealkylase activity in rat liver microsomes was markedly inhibited by PBYO and PBO, partly by PBYOL, but not by PBA. PMID- 11901096 TI - Distribution of brimonidine into anterior and posterior tissues of monkey, rabbit, and rat eyes. AB - The objectives of the study were to evaluate the distribution of brimonidine (alpha2-adrenergic agonist) into anterior and posterior ocular tissues. Single or multiple doses of a 0.2 or 0.5% brimonidine tartrate solution were administered to one or both eyes of monkeys or to one eye of rabbits. Brimonidine was administered intraperitoneally to rats. After topical administration, [14C]brimonidine was rapidly absorbed into the cornea and conjunctiva and distributed throughout the eye. [14C]Radioactivity was higher and cleared more slowly in pigmented tissues (iris/ciliary body, choroid/retina, and optic nerve) than in nonpigmented tissues. Single and multiple dosing led to a similar drug distribution, with higher levels of brimonidine measured in pigmented tissues after multiple dosing. Most of the radioactivity extracted from ocular tissues represented unchanged brimonidine. In the rabbits and the monkey treated in only one eye, levels of radioactivity in the untreated eye were low, consistent with the low systemic levels and rapid drug clearance. Posterior ocular tissue concentrations of radioactivity exceeded systemic blood concentrations. The vitreous humor brimonidine concentrations in monkeys treated topically with 0.2% brimonidine tartrate was 82 +/- 45 nM. Vitreous levels in rabbits confirmed the penetration of brimonidine to the posterior segment. Similar concentrations of brimonidine (22 to 390 nM) were measured in the vitreous and retina of rats injected intraperitoneally with brimonidine. Both topically applied and systemically administered brimonidine reach the back of the eye at nanomolar concentrations sufficient to activate alpha2-adrenergic receptors. The brimonidine levels achieved at the retina are relevant for neuroprotection models. PMID- 11901097 TI - Disposition of the selective cholesterol absorption inhibitor ezetimibe in healthy male subjects. AB - Ezetimibe [SCH 58235; 1-(4-fluorophenyl)-3(R)-[3-(4-fluorophenyl)-3(S) hydroxypropyl]-4(S)-(4-hydroxyphenyl)-2-azetidinone], a selective cholesterol absorption inhibitor, is being developed for the treatment of primary hypercholesterolemia. The absorption, metabolism, and excretion of ezetimibe were characterized in eight healthy male volunteers in this single-center, single dose, open-label study. Subjects received a single oral 20-mg dose of [14C]ezetimibe (approximately 100 microCi) with 200 ml of noncarbonated water after a 10-h fast. Concentrations of radioactivity and/or ezetimibe (conjugated and unconjugated) were determined in plasma, urine, and fecal samples. Ezetimibe was rapidly absorbed and extensively conjugated following oral administration. The main circulating metabolite in plasma was SCH 60663 [1-O-[4-[trans-(2S,3R)-1 (4-fluorophenyl)-4-oxo-3-[3(S)-hydroxy-3-(4-fluorophenyl)propyl]-2 azetidinyl]phenyl]-beta-D-glucuronic acid], the glucuronide conjugate of ezetimibe. Plasma concentration-time profiles of unconjugated and conjugated drug exhibited multiple peaks, indicating enterohepatic recycling. Approximately 78 and 11% of the administered [14C]ezetimibe dose were excreted in feces and urine, respectively, by 240 h after drug administration. Total recovery of radioactivity averaged 89% of the administered dose. The main excreted metabolite was the glucuronide conjugate of ezetimibe. The primary metabolite in urine (0- to72-h composite) was also the glucuronide conjugate (about 9% of the administered dose). Significant amounts (69% of the dose) of ezetimibe were present in the feces, presumably as a result of SCH 60663 hydrolysis and/or unabsorbed drug. No adverse events were reported in this study. A single 20-mg capsule of [(14)C]ezetimibe was safe and well tolerated after oral administration. The pharmacokinetics of ezetimibe are consistent with extensive glucuronidation and enterohepatic recirculation. The primary metabolic pathway for ezetimibe is by glucuronidation of the 4-hydroxyphenyl group. PMID- 11901098 TI - Regioselective metabolism of taxoids by human CYP3A4 and 2C8: structure-activity relationship. AB - Paclitaxel and docetaxel are metabolized by liver microsomal monooxygenases into inactive metabolites further eliminated from the body via the bile route. In spite of their close chemical structure, the two drugs are oxidized by two different enzymes; CYP2C8 catalyzes the 6-hydroxylation on the taxane ring of paclitaxel, whereas CYP3A4 oxidizes docetaxel on the tert-butyl group of the lateral chain in C13. Since paclitaxel and docetaxel differ only by two substitutions, the role of individual modifications was investigated; the regioselectivity of hydroxylation was assessed by high-pressure liquid chromatography/mass spectrometry, and enzymes implicated in individual reactions were identified using human liver microsomes and recombinant P450 expressed in Ad293 cells. The biotransformation of docetaxel, 10-deacetylpaclitaxel, and 10 deacetylbaccatin III was steadily increased (2- to 5-fold) by the addition of an acetyl group in position 10, suggesting that the presence of a hydrophobic group in position 10 stimulated hydroxylation by P450 proteins. The absence of the lateral chain at C13 in baccatin III severely impaired the metabolism supported by CYP3A4. The presence of a tert-butyl group in the lateral chain of docetaxel favored the hydroxylation on the tert-butyl by CYP3A4, whereas the presence of a phenyl group in the lateral chain facilitated the oxidation on the taxane ring by CYP2C8. Collectively, these data strongly suggested that the structure of the lateral chain and the nature of substituent in position 10 play an important role in determining the regioselective oxidation by P450 proteins and modulate the reaction rate by human liver microsomes. PMID- 11901100 TI - CYP3A4 active site volume modification by mutagenesis of leucine 211. AB - The leucine 211 --> phenylalanine (L211F) and leucine 211 --> tyrosine (L211Y) mutant forms of cytochrome P450 3A4 have been generated by site-directed mutagenesis and expressed functionally in Escherichia coli. Substrate binding affinities (S50 values) for testosterone and 7-benzyloxy-4 trifluoromethylcoumarin (BFC) were similar for the mutants and wild-type CYP3A4 (49 and 21 microM for L211F, 35 and 20 microM for L211Y, and 33 and 20 microM for the wild type, respectively). For erythromycin, however, the K(m) values determined for the L211F and L211Y mutants were 2.4- and 10.5-fold higher than for the wild type. Furthermore, IC50 values for the inhibition of testosterone 6 beta-hydroxylation by erythromycin and troleandomycin for L211F were 2.4- and 3.7 fold higher, and those for L211Y were 3.4- and 9.2-fold higher than those measured for the wild type. Conversely, small inhibitors, such as diazepam, exhibited no significant difference in IC50 values between the wild type and the L211F and L211Y mutants. It is proposed that large substrates bound in the catalytic center of CYP3A4 with molecular volumes greater than approximately 600 A(3) were less well accommodated in the altered active sites, resulting in lower association energies and increased IC50 values. PMID- 11901099 TI - Determination of acetaminophen-protein adducts in mouse liver and serum and human serum after hepatotoxic doses of acetaminophen using high-performance liquid chromatography with electrochemical detection. AB - Acetaminophen-induced hepatotoxicity has been attributed to covalent binding of the reactive metabolite N-acetyl-p-benzoquinone imine to cysteine groups on proteins as an acetaminophen-cysteine conjugate. We report a high-performance liquid chromatography with electrochemical detection (HPLC-ECD) assay for the conjugate with increased sensitivity compared with previous methods. Previous methods to quantitate the protein-bound conjugate have used a competitive immunoassay or radiolabeled acetaminophen. With HPLC-ECD, the protein samples are dialyzed and then digested with protease. The acetaminophen-cysteine conjugate is then quantified by HPLC-ECD using tyrosine as an internal reference. The lower limit of detection of the assay is approximately 3 pmol/mg of protein. Acetaminophen protein adducts were detected in liver and serum as early as 15 min after hepatotoxic dosing of acetaminophen to mice. Adducts were also detected in the serum of acetaminophen overdose patients. Analysis of human serum samples for the acetaminophen-cysteine conjugate revealed a positive correlation between acetaminophen-cysteine conjugate concentration and serum aspartate aminotransferase (AST) activity or time. Adducts were detected in the serum of patients even with relatively mild liver injury, as measured by AST and alanine aminotransferase. This assay may be useful in the diagnostic evaluation of patients with hepatotoxicity of an indeterminate etiology for which acetaminophen toxicity is suspect. PMID- 11901101 TI - Delineating the contribution of secretory transporters in the efflux of etoposide using Madin-Darby canine kidney (MDCK) cells overexpressing P-glycoprotein (Pgp), multidrug resistance-associated protein (MRP1), and canalicular multispecific organic anion transporter (cMOAT). AB - Multidrug resistance conferred to cancer cells is often mediated by the expression of efflux transporter "pumps". It is also believed that many of the same transporters are involved in drug efflux from numerous normal endothelial and epithelial cell types in the intestine, brain, kidney, and liver. Etoposide transport kinetics were characterized in Caco-2 cells and in well established Madin-Darby canine kidney (MDCKII) cell lines that were stably-transfected with a human cDNA encoding P-glycoprotein (Pgp), human multidrug resistance protein (MRP1), or the canalicular multispecific organic anion (cMOAT) transporters to determine the roles of these transporters in etoposide efflux. Etoposide transport kinetics were concentration-dependent in the MDCKII-MDR1 and MDCKII cMOAT cells. The apparent secretory Michaelis constant (Km) and carrier-mediated permeability (Pc) values for Pgp and cMOAT were 254.96 +/- 94.39 microM and 5.96 +/- 0.41 x 10(-6) cm/s and 616.54 +/- 163.15 microM and 1.87 +/- 0.10 x 10(-5) cm/s, respectively. The secretory permeability of etoposide decreased significantly in the basal to apical (B to A) (i.e., efflux) direction, whereas the permeability increased 2.3-fold in the apical to basal (A to B) direction in MDCKII-MDR1 cells in the presence of elacridar (GF120918). Moderate inhibition of etoposide efflux by leukotriene C4 (LTC4) was observed in MDCKII-cMOAT cells. Furthermore, etoposide inhibited LTC4 efflux, confirming the involvement of cMOAT. The flux of etoposide in MDCKII-MRP1 cells was similar to that in MDCKII/wt control cells. The current results demonstrate that the secretory transport mechanism of etoposide involves multiple transporters, including Pgp and cMOAT but not MRP1. These results demonstrate that Pgp and cMOAT are involved in the intestinal secretory transport of etoposide. Since the intestinal secretion of etoposide was previously reported in the literature, it also suggests that they may be involved in the in vivo intestinal secretion of etoposide; however, mechanistic in vivo studies are required to confirm this. PMID- 11901102 TI - The oxidative biotransformation of losoxantrone (CI-941). AB - The oxidative biotransformation of the anticancer drug 7-hydroxy-2-[2-[(2 hydroxyethyl)amino]ethyl]-5-[[2-[(2-hydroxyethyl)amino]ethyl]amino]anthra[1,9 cd]pyrazol-6(2H)-one dihydrochloride (losoxantrone, CI-941) after incubation of primary cultures of rat hepatocytes has been investigated. The structures of twelve losoxantrone metabolites have been elucidated by means of high-performance liquid chromatography-mass spectometry, tandem mass spectrometry, and two dimensional NMR. In these mammalian hepatocytes, the CI-941 biotransformation includes a monohydroxylation of the phenolic substructure of the CI-941 chromophore via cytochrome P450 catalysis, resulting in metabolites having an ortho- and para-hydroquinonoid substructure, respectively. The identification of a glutathione conjugate as a follow-up metabolite confirms the oxidative activation of the ortho-hydroxylated losoxantrone metabolite. The oxidative activation establishes the ability of CI-941 to form covalent bonds to intracellular nucleophilic targets. Furthermore, the CI-941 metabolism was shown to be extremely suppressed in rat hepatocytes incubated with metyrapone. In contrast to these results, human tumor HepG2 cells did not show any CI-941 biotransformation after incubation. PMID- 11901103 TI - Oliver Nelson and quality protein maize. PMID- 11901104 TI - Predicting evolutionary potential: in vitro evolution accurately reproduces natural evolution of the tem beta-lactamase. AB - To evaluate the validity of our in vitro evolution method as a model for natural evolutionary processes, the TEM-1 beta-lactamase gene was evolved in vitro and was selected for increased resistance to cefotaxime, cefuroxime, ceftazadime, and aztreonam, i.e., the "extended-spectrum" phenotype. The amino acid substitutions recovered in 10 independent in vitro evolvants were compared with the amino acid substitutions in the naturally occurring extended-spectrum TEM alleles. Of the nine substitutions that have arisen multiple times in naturally occurring extended-spectrum TEM alleles, seven were recovered multiple times in vitro. We take this result as evidence that our in vitro evolution technique accurately mimics natural evolution and can therefore be used to predict the results of natural evolutionary processes. Additionally, our results predict that a phenotype not yet observed among TEM beta-lactamases in nature-resistance to cefepime-is likely to arise in nature. PMID- 11901105 TI - Geographic uniformity of the Lyme disease spirochete (Borrelia burgdorferi) and its shared history with tick vector (Ixodes scapularis) in the Northeastern United States. AB - Over 80% of reported cases of Lyme disease in the United States occur in coastal regions of northeastern and mid-Atlantic states. The genetic structure of the Lyme disease spirochete (Borrelia burgdorferi) and its main tick vector (Ixodes scapularis) was studied concurrently and comparatively by sampling natural populations of I. scapularis ticks along the East Coast from 1996 to 1998. Borrelia is genetically highly diverse at the outer surface protein ospC. Since Borrelia is highly clonal, the ospC alleles can be used to define clones. A newly designed reverse line blotting (RLB) assay shows that up to 10 Borrelia clones can infect a single tick. The clone frequencies in Borrelia populations are the same across the Northeast. On the other hand, I. scapularis populations show strong regional divergence (among northeastern, mid-Atlantic, and southern states) as well as local differentiation. The high genetic diversity within Borrelia populations and the disparity in the genetic structure between Borrelia and its tick vector are likely consequences of strong balancing selection on local Borrelia clones. Demographically, both Borrelia and I. scapularis populations in the Northeast show the characteristics of a species that has recently expanded from a population bottleneck. Major geological and ecological events, such as the last glacial maximum (18,000 years ago) and the modern-day expansion of tick habitats, are likely causes of the observed "founder effects" for the two organisms in the Northeast. We therefore conclude that the genetic structure of B. burgdorferi has been intimately shaped by the natural history of its main vector, the northern lineage of I. scapularis ticks. PMID- 11901106 TI - Crossing over between regions of limited homology in Escherichia coli. RecA dependent and RecA-independent pathways. AB - We have developed an assay for intermolecular crossing over between circular plasmids carrying variable amounts of homology. Screens of Escherichia coli mutants demonstrated that known recombination functions can only partially account for the observed recombination. Recombination rates increased three to four orders of magnitude as homology rose from 25 to 411 bp. Loss of recA blocked most recombination; however, RecA-independent crossing over predominated at 25 bp and could be detected at all homology lengths. Products of recA-independent recombination were reciprocal in nature. This suggests that RecA-independent recombination may involve a true break-and-join mechanism, but the genetic basis for this mechanism remains unknown. RecA-dependent crossing over occurred primarily by the RecF pathway but considerable recombination occurred independent of both RecF and RecBCD. In many respects, the genetic dependence of RecA dependent crossing over resembled that reported for single-strand gap repair. Surprisingly, ruvC mutants, in both recA(+) and recA mutant backgrounds, scored as hyperrecombinational. This may occur because RuvC preferentially resolves Holliday junction intermediates, critical to both RecA-dependent and RecA independent mechanisms, to the noncrossover configuration. Levels of crossing over were increased by defects in DnaB helicase and by oxidative damage, showing that damaged DNA or stalled replication can initiate genetic recombination. PMID- 11901107 TI - Fission yeast mutants affecting telomere clustering and meiosis-specific spindle pole body integrity. AB - In meiotic prophase of many eukaryotic organisms, telomeres attach to the nuclear envelope and form a polarized configuration called the bouquet. Bouquet formation is hypothesized to facilitate homologous chromosome pairing. In fission yeast, bouquet formation and telomere clustering occurs in karyogamy and persists throughout the horsetail stage. Here we report the isolation and characterization of six mutants from our screen for meiotic mutants. These mutants show defective telomere clustering as demonstrated by mislocalization of Swi6::GFP, a heterochromatin-binding protein, and Taz1p::GFP, a telomere-specific protein. These mutants define four complementation groups and are named dot1 to dot4 defective organization of telomeres. dot3 and dot4 are allelic to mat1-Mm and mei4, respectively. Immunolocalization of Sad1, a protein associated with the spindle pole body (SPB), in dot mutants showed an elevated frequency of multiple Sad1-nuclei signals relative to wild type. Many of these Sad1 foci were colocalized with Taz1::GFP. Impaired SPB structure and function were further demonstrated by failure of spore wall formation in dot1, by multiple Pcp1::GFP signals (an SPB component) in dot2, and by abnormal microtubule organizations during meiosis in dot mutants. The coincidence of impaired SPB functions with defective telomere clustering suggests a link between the SPB and the telomere cluster. PMID- 11901108 TI - Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD(+) salvage pathway. AB - The Sir2 protein is an NAD(+)-dependent protein deacetylase that is required for silencing at the silent mating-type loci, telomeres, and the ribosomal DNA (rDNA). Mutations in the NAD(+) salvage gene NPT1 weaken all three forms of silencing and also cause a reduction in the intracellular NAD(+) level. We now show that mutation of a highly conserved histidine residue in Npt1p results in a silencing defect, indicating that Npt1p enzymatic activity is required for silencing. Deletion of another NAD(+) salvage pathway gene called PNC1 caused a less severe silencing defect and did not significantly reduce the intracellular NAD(+) concentration. However, silencing in the absence of PNC1 was completely dependent on the import of nicotinic acid from the growth medium. Deletion of a gene in the de novo NAD(+) synthesis pathway BNA1 resulted in a significant rDNA silencing defect only on medium deficient in nicotinic acid, an NAD(+) precursor. By immunofluorescence microscopy, Myc-tagged Bna1p was localized throughout the whole cell in an asynchronously growing population. In contrast, Myc-tagged Npt1p was highly concentrated in the nucleus in approximately 40% of the cells, indicating that NAD(+) salvage occurs in the nucleus in a significant fraction of cells. We propose a model in which two components of the NAD(+) salvage pathway, Pnc1p and Npt1p, function together in recycling the nuclear nicotinamide generated by Sir2p deacetylase activity back into NAD(+). PMID- 11901109 TI - UV irradiation causes the loss of viable mitotic recombinants in Schizosaccharomyces pombe cells lacking the G(2)/M DNA damage checkpoint. AB - Elevated mitotic recombination and cell cycle delays are two of the cellular responses to UV-induced DNA damage. Cell cycle delays in response to DNA damage are mediated via checkpoint proteins. Two distinct DNA damage checkpoints have been characterized in Schizosaccharomyces pombe: an intra-S-phase checkpoint slows replication and a G(2)/M checkpoint stops cells passing from G(2) into mitosis. In this study we have sought to determine whether UV damage-induced mitotic intrachromosomal recombination relies on damage-induced cell cycle delays. The spontaneous and UV-induced recombination phenotypes were determined for checkpoint mutants lacking the intra-S and/or the G(2)/M checkpoint. Spontaneous mitotic recombinants are thought to arise due to endogenous DNA damage and/or intrinsic stalling of replication forks. Cells lacking only the intra-S checkpoint exhibited no UV-induced increase in the frequency of recombinants above spontaneous levels. Mutants lacking the G(2)/M checkpoint exhibited a novel phenotype; following UV irradiation the recombinant frequency fell below the frequency of spontaneous recombinants. This implies that, as well as UV-induced recombinants, spontaneous recombinants are also lost in G(2)/M mutants after UV irradiation. Therefore, as well as lack of time for DNA repair, loss of spontaneous and damage-induced recombinants also contributes to cell death in UV-irradiated G(2)/M checkpoint mutants. PMID- 11901110 TI - Analysis of conditional mutations in the Saccharomyces cerevisiae MLH1 gene in mismatch repair and in meiotic crossing over. AB - In mismatch repair (MMR), members of the MLH gene family have been proposed to act as key molecular matchmakers to coordinate mismatch recognition with downstream repair functions that result in mispair excision. Two members of this gene family, MLH1 and MLH3, have also been implicated in meiotic crossing over. These diverse roles suggest that a mutational analysis of MLH genes could provide reagents required to identify interactions between gene products and to test whether the different roles ascribed to a subset of these genes can be separated. In this report we show that in Saccharomyces cerevisiae the mlh1Delta mutation confers inviability in pol3-01 strain backgrounds that are defective in the Poldelta proofreading exonuclease activity. This phenotype was exploited to identify four mlh1 alleles that each confer a temperature-sensitive phenotype for viability in pol3-01 strains. In three different mutator assays, strains bearing conditional mlh1 alleles displayed wild-type or nearly wild-type mutation rates at 26 degrees. At 35 degrees, these strains exhibited mutation rates that approached those observed in mlh1Delta mutants. The mutator phenotype exhibited in mlh1-I296S strains was partially suppressed at 35 degrees by EXO1 overexpression. The mlh1-F228S and -I296S mutations conferred a separation-of function phenotype in meiosis; both mlh1-F228S and -I296S strains displayed strong defects in meiotic mismatch repair but showed nearly wild-type levels of crossing over, suggesting that the conditional mutations differentially affected MLH1 functions. These genetic studies suggest that the conditional mlh1 mutations can be used to separate the MMR and meiotic crossing-over functions of MLH1 and to identify interactions between MLH1 and downstream repair components. PMID- 11901112 TI - Isolation and characterization of the Cryptococcus neoformans MATa pheromone gene. AB - Cryptococcus neoformans is a heterothallic basidiomycete with two mating types, MATa and MATalpha. The mating pathway of this fungus has a number of conserved genes, including a MATalpha-specific pheromone (MFalpha1). A modified differential display strategy was used to identify a gene encoding the MATa pheromone. The gene, designated MFa1, is 42 amino acids in length and contains a conserved farnesylation motif. MFa1 is present in three linked copies that span a 20-kb fragment of MATa-specific DNA and maps to the MAT-containing chromosome. Transformation studies showed that MFa1 induced filament formation only in MATalpha cells, demonstrating that MFa1 is functionally conserved. Sequence analysis of the predicted Mfa1 and Mfalpha1 proteins revealed that, in contrast to other fungi such as Saccharomyces cerevisiae, the C. neoformans pheromone genes are structurally and functionally conserved. However, unlike the MFalpha1 gene, which is found in MATalpha strains of both varieties of C. neoformans, MFa1 is specific for the neoformans variety of C. neoformans. PMID- 11901111 TI - The novel adaptor protein, Mti1p, and Vrp1p, a homolog of Wiskott-Aldrich syndrome protein-interacting protein (WIP), may antagonistically regulate type I myosins in Saccharomyces cerevisiae. AB - Type I myosins in yeast, Myo3p and Myo5p (Myo3/5p), are involved in the reorganization of the actin cytoskeleton. The SH3 domain of Myo5p regulates the polymerization of actin through interactions with both Las17p, a homolog of mammalian Wiskott-Aldrich syndrome protein (WASP), and Vrp1p, a homolog of WASP interacting protein (WIP). Vrp1p is required for both the localization of Myo5p to cortical patch-like structures and the ATP-independent interaction between the Myo5p tail region and actin filaments. We have identified and characterized a new adaptor protein, Mti1p (Myosin tail region-interacting protein), which interacts with the SH3 domains of Myo3/5p. Mti1p co-immunoprecipitated with Myo5p and Mti1p GFP co-localized with cortical actin patches. A null mutation of MTI1 exhibited synthetic lethal phenotypes with mutations in SAC6 and SLA2, which encode actin bundling and cortical actin-binding proteins, respectively. Although the mti1 null mutation alone did not display any obvious phenotype, it suppressed vrp1 mutation phenotypes, including temperature-sensitive growth, abnormally large cell morphology, defects in endocytosis and salt-sensitive growth. These results suggest that Mti1p and Vrp1p antagonistically regulate type I myosin functions. PMID- 11901113 TI - Genetic and physical mapping of Avr1a in Phytophthora sojae. AB - The interaction between soybean and the phytopathogenic oomycete Phytophthora sojae is controlled by host resistance (Rps) genes and pathogen avirulence (Avr) genes. We have mapped the Avr1a locus in F(2) populations derived from four different P. sojae races. Four RAPD and nine AFLP markers linked to Avr1a were initially identified. Nine markers were used to compare genetic linkage maps of the Avr1a locus in two distinct F(2) populations. Distorted segregation ratios favoring homozygous genotypes were noted in both crosses. Segregation analysis of all the markers in one F(2) population of 90 progeny generated a map of 113.2 cM encompassing Avr1a, with one marker cosegregating with the gene. The cosegregating DNA marker was used to isolate P. sojae BAC clones and construct a physical map covering 170 kb, from which additional DNA markers were developed. Three markers occurring within the BAC contig were mapped in an enlarged population of 486 F(2) progeny. Avr1a was localized to a 114-kb interval, and an average physical to genetic distance ratio of 391 kb/cM was calculated for this region. This work provides a basis for the positional cloning of Avr1a. PMID- 11901115 TI - Selection and maintenance of androdioecy in Caenorhabditis elegans. AB - Caenorhabditis elegans is an androdioecious nematode composed of selfing hermaphrodites and rare males. A model of male maintenance demonstrates that selfing rates in hermaphrodites cannot be too high or else the frequency of males will be driven down to the rate of spontaneous nondisjunction of the X chromosome. After their outcrossing ability is assessed, males are found to skirt the frequency range in which they would be maintained. When male maintenance is directly assessed by elevating male frequency and observing the frequency change through time, males are gradually eliminated from the population. Males, therefore, appear to reproduce at a rate just below that necessary for them to be maintained. Populations polymorphic for a mutation (fog-2) that effectively changes hermaphrodites into females demonstrate that there is strong selection against dioecy. Factors such as variation in male mating ability and inbreeding depression could potentially lead to the long-term maintenance of males. PMID- 11901114 TI - Identification and characterization of the genes encoding the core histones and histone variants of Neurospora crassa. AB - We have identified and characterized the complete complement of genes encoding the core histones of Neurospora crassa. In addition to the previously identified pair of genes that encode histones H3 and H4 (hH3 and hH4-1), we identified a second histone H4 gene (hH4-2), a divergently transcribed pair of genes that encode H2A and H2B (hH2A and hH2B), a homolog of the F/Z family of H2A variants (hH2Az), a homolog of the H3 variant CSE4 from Saccharomyces cerevisiae (hH3v), and a highly diverged H4 variant (hH4v) not described in other species. The hH4-1 and hH4-2 genes, which are 96% identical in their coding regions and encode identical proteins, were inactivated independently. Strains with inactivating mutations in either gene were phenotypically wild type, in terms of growth rates and fertility, but the double mutants were inviable. As expected, we were unable to isolate null alleles of hH2A, hH2B, or hH3. The genomic arrangement of the histone and histone variant genes was determined. hH2Az and the hH3-hH4-1 gene pair are on LG IIR, with hH2Az centromere-proximal to hH3-hH4-1 and hH3 centromere-proximal to hH4-1. hH3v and hH4-2 are on LG IIIR with hH3v centromere proximal to hH4-2. hH4v is on LG IVR and the hH2A-hH2B pair is located immediately right of the LG VII centromere, with hH2A centromere-proximal to hH2B. Except for the centromere-distal gene in the pairs, all of the histone genes are transcribed toward the centromere. Phylogenetic analysis of the N. crassa histone genes places them in the Euascomycota lineage. In contrast to the general case in eukaryotes, histone genes in euascomycetes are few in number and contain introns. This may be a reflection of the evolution of the RIP (repeat induced point mutation) and MIP (methylation induced premeiotically) processes that detect sizable duplications and silence associated genes. PMID- 11901116 TI - Why are there males in the hermaphroditic species Caenorhabditis elegans? AB - The free-living nematode worm Caenorhabditis elegans reproduces primarily as a self-fertilizing hermaphrodite, yet males are maintained in wild-type populations at low frequency. To determine the role of males in C. elegans, we develop a mathematical model for the genetic system of hermaphrodites that can either self fertilize or be fertilized by males and we perform laboratory observations and experiments on both C. elegans and a related dioecious species C. remanei. We show that the mating efficiency of C. elegans is poor compared to a dioecious species and that C. elegans males are more attracted to C. remanei females than they are to their conspecific hermaphrodites. We postulate that a genetic mutation occurred during the evolution of C. elegans hermaphrodites, resulting in the loss of an attracting sex pheromone present in the ancestor of both C. elegans and C. remanei. Our findings suggest that males are maintained in C. elegans because of the particular genetic system inherited from its dioecious ancestor and because of nonadaptive spontaneous nondisjunction of sex chromosomes, which occurs during meiosis in the hermaphrodite. A theoretical argument shows that the low frequency of male mating observed in C. elegans can support male-specific genes against mutational degeneration. This results in the continuing presence of functional males in a 99.9% hermaphroditic species in which outcrossing is disadvantageous to hermaphrodites. PMID- 11901118 TI - Timing and targeting of P-element local transposition in the male germline cells of Drosophila melanogaster. AB - The P element in Drosophila melanogaster preferentially transposes into nearby sites. The local insertions display a preferential orientation toward the starting element. We investigated the mechanism of the P-element local transposition by isolating and characterizing local insertions in the male germline. We designed a genetic screen employing a marker gene that is carried in the P element and is dose sensitive. This dose effect allows isolation of flies containing newly transposed P elements in the presence of the starting element. A rapid molecular screen with PCR was used to identify 45 local insertions located within an approximately 40-kb genomic region on both sides of the starting element. Our system permits the isolation of the cluster progeny derived from a single insertion event, but none was isolated. The data suggest that local transposition occurs in the meiotic cell cycle. Nearly all of the local insertions were located within the promoter regions of the genes that were active in the male germline cells, suggesting that local insertions target predominantly active promoters. Our analysis shows that local transposition of the P element is highly regulated, displaying a cell-type specificity and a target specificity. PMID- 11901117 TI - Modifiers of terminal deficiency-associated position effect variegation in Drosophila. AB - Terminal deletions of a Drosophila minichromosome (Dp(1;f)1187) dramatically increase the position effect variegation (PEV) of a yellow(+) body-color gene located in cis. Such terminal deficiency-associated PEV (TDA-PEV) can be suppressed by the presence of a second minichromosome, a phenomenon termed "trans suppression." We performed a screen for mutations that modify TDA-PEV and trans suppression. Seventy suppressors and enhancers of TDA-PEV were identified, but no modifiers of trans-suppression were recovered. Secondary analyses of the effects of these mutations on different PEV types identified 10 mutations that modify only TDA-PEV and 6 mutations that modify TDA-PEV and only one other type of PEV. One mutation, a new allele of Su(var)3-9, affects all forms of PEV, including silencing associated with the insertion of a transgene into telomeric regions (TPE). This Su(var)3-9 allele is the first modifier of PEV to affect TPE and provides a unique link between different types of gene silencing in Drosophila. The remaining mutations affected multiple PEV types, indicating that general PEV modifiers impact TDA-PEV. Modifiers of TDA-PEV may identify proteins that play important roles in general heterochromatin biology, including proteins involved in telomere structure and function and the organization of chromosomes in the interphase nucleus. PMID- 11901119 TI - The Drosophila suppressor of underreplication protein binds to late-replicating regions of polytene chromosomes. AB - In many late-replicating euchromatic regions of salivary gland polytene chromosomes, DNA is underrepresented. A mutation in the SuUR gene suppresses underreplication and leads to normal levels of DNA polytenization in these regions. We identified the SuUR gene and determined its structure. In the SuUR mutant stock a 6-kb insertion was found in the fourth exon of the gene. A single SuUR transcript is present at all stages of Drosophila development and is most abundant in adult females and embryos. The SuUR gene encodes a protein of 962 amino acids whose putative sequence is similar to the N-terminal part of SNF2/SWI2 proteins. Staining of salivary gland polytene chromosomes with antibodies directed against the SuUR protein shows that the protein is localized mainly in late-replicating regions and in regions of intercalary and pericentric heterochromatin. PMID- 11901120 TI - A misexpression study examining dorsal thorax formation in Drosophila melanogaster. AB - We studied thorax formation in Drosophila melanogaster using a misexpression screen with EP lines and thoracic Gal4 drivers that provide a genetically sensitized background. We identified 191 interacting lines showing alterations of thoracic bristles (number and/or location), thorax and scutellum malformations, lethality, or suppression of the thoracic phenotype used in the screen. We analyzed these lines and showed that known genes with different functional roles (selector, prepattern, proneural, cell cycle regulation, lineage restriction, signaling pathways, transcriptional control, and chromatin organization) are among the modifier lines. A few lines have previously been identified in thorax formation, but others, such as chromatin-remodeling complex genes, are novel. However, most of the interacting loci are uncharacterized, providing a wealth of new genetic data. We also describe one such novel line, poco pelo (ppo), where both misexpression and loss-of-function phenotypes are similar: loss of bristles and scutellum malformation. PMID- 11901122 TI - Sequence of the Tribolium castaneum homeotic complex: the region corresponding to the Drosophila melanogaster antennapedia complex. AB - The homeotic selector genes of the red flour beetle, Tribolium castaneum, are located in a single cluster. We have sequenced the region containing the homeotic selector genes required for proper development of the head and anterior thorax, which is the counterpart of the ANTC in Drosophila. This 280-kb interval contains eight homeodomain-encoding genes, including single orthologs of the Drosophila genes labial, proboscipedia, Deformed, Sex combs reduced, fushi tarazu, and Antennapedia, as well as two orthologs of zerknullt. These genes are all oriented in the same direction, as are the Hox genes of amphioxus, mice, and humans. Although each transcription unit is similar to its Drosophila counterpart in size, the Tribolium genes contain fewer introns (with the exception of the two zerknullt genes), produce shorter mRNAs, and encode smaller proteins. Unlike the ANTC, this region of the Tribolium HOMC contains no additional genes. PMID- 11901121 TI - Genetic and molecular analysis of region 88E9;88F2 in Drosophila melanogaster, including the ear gene related to human factors involved in lineage-specific leukemias. AB - We identified and characterized the Drosophila gene ear (ENL/AF9-related), which is closely related to mammalian genes that have been implicated in the onset of acute lymphoblastic and myelogenous leukemias when their products are fused as chimeras with those of human HRX, a homolog of Drosophila trithorax. The ear gene product is present in all early embryonic cells, but becomes restricted to specific tissues in late embryogenesis. We mapped the ear gene to cytological region 88E11-13, near easter, and showed that it is deleted by Df(3R)ea(5022rx1), a small, cytologically invisible deletion. Annotation of the completed Drosophila genome sequence suggests that this region might contain as many as 26 genes, most of which, including ear, are not represented by mutant alleles. We carried out a large-scale noncomplementation screen using Df(3R)ea(5022rx1) and chemical (EMS) mutagenesis from which we identified seven novel multi-allele recessive lethal complementation groups in this region. An overlapping deficiency, Df(3R)Po(4), allowed us to map several of these groups to either the proximal or the distal regions of Df(3R)ea(5022rx1). One of these complementation groups likely corresponds to the ear gene as judged by map location, terminal phenotype, and reduction of EAR protein levels. PMID- 11901123 TI - History of infection with different male-killing bacteria in the two-spot ladybird beetle Adalia bipunctata revealed through mitochondrial DNA sequence analysis. AB - The two-spot ladybird beetle Adalia bipunctata (Coleoptera: Coccinellidae) is host to four different intracellular maternally inherited bacteria that kill male hosts during embryogenesis: one each of the genus Rickettsia (alpha Proteobacteria) and Spiroplasma (Mollicutes) and two distinct strains of Wolbachia (alpha-Proteobacteria). The history of infection with these male killers was explored using host mitochondrial DNA, which is linked with the bacteria due to joint maternal inheritance. Two variable regions, 610 bp of cytochrome oxidase subunit I and 563 bp of NADH dehydrogenase subunit 5, were isolated from 52 A. bipunctata with known infection status and different geographic origin from across Eurasia. Two outgroup taxa were also considered. DNA sequence analysis revealed that the distribution of mitochondrial haplotypes is not associated with geography. Rather, it correlates with infection status, confirming linkage disequilibrium between mitochondria and bacteria. The data strongly suggest that the Rickettsia male-killer invaded the host earlier than the other taxa. Further, the male-killing Spiroplasma is indicated to have undergone a recent and extensive spread through host populations. In general, male-killing in A. bipunctata seems to represent a highly dynamic system, which should prove useful in future studies on the evolutionary dynamics of this peculiar type of symbiont-host association. PMID- 11901124 TI - Mutualistic Wolbachia infection in Aedes albopictus: accelerating cytoplasmic drive. AB - Maternally inherited rickettsial symbionts of the genus Wolbachia occur commonly in arthropods, often behaving as reproductive parasites by manipulating host reproduction to enhance the vertical transmission of infections. One manipulation is cytoplasmic incompatibility (CI), which causes a significant reduction in brood hatch and promotes the spread of the maternally inherited Wolbachia infection into the host population (i.e., cytoplasmic drive). Here, we have examined a Wolbachia superinfection in the mosquito Aedes albopictus and found the infection to be associated with both cytoplasmic incompatibility and increased host fecundity. Relative to uninfected females, infected females live longer, produce more eggs, and have higher hatching rates in compatible crosses. A model describing Wolbachia infection dynamics predicts that increased fecundity will accelerate cytoplasmic drive rates. To test this hypothesis, we used population cages to examine the rate at which Wolbachia invades an uninfected Ae. albopictus population. The observed cytoplasmic drive rates were consistent with model predictions for a CI-inducing Wolbachia infection that increases host fecundity. We discuss the relevance of these results to both the evolution of Wolbachia symbioses and proposed applied strategies for the use of Wolbachia infections to drive desired transgenes through natural populations (i.e., population replacement strategies). PMID- 11901125 TI - Precocious expression of the Glide/Gcm glial-promoting factor in Drosophila induces neurogenesis. AB - Neurons and glial cells depend on similar developmental pathways and often originate from common precursors; however, the differentiation of one or the other cell type depends on the activation of cell-specific pathways. In Drosophila, the differentiation of glial cells depends on a transcription factor, Glide/Gcm. This glial-promoting factor is both necessary and sufficient to induce the central and peripheral glial fates at the expense of the neuronal fate. In a screen for mutations affecting the adult peripheral nervous system, we have found a dominant mutation inducing supernumerary sensory organs. Surprisingly, this mutation is allelic to glide/gcm and induces precocious glide/gcm expression, which, in turn, activates the proneural genes. As a consequence, sensory organs are induced. Thus, temporal misregulation of the Glide/Gcm glial-promoting factor reveals a novel potential for this cell fate determinant. At the molecular level, this implies unpredicted features of the glide/gcm pathway. These findings also emphasize the requirement for both spatial and temporal glide/gcm regulation to achieve proper cell specification within the nervous system. PMID- 11901126 TI - Multiple effects of genetic background on variegated transgene expression in mice. AB - BLG/7 transgenic mice express an ovine beta-lactoglobulin transgene during lactation. Unusually, transgene expression levels in milk differ between siblings. This variable expression is due to variegated transgene expression in the mammary gland and is reminiscent of position-effect variegation. The BLG/7 line was created and maintained on a mixed CBA x C57BL/6 background. We have investigated the effect on transgene expression of backcrossing for 13 generations into these backgrounds. Variable transgene expression was observed in all populations examined, confirming that it is an inherent property of the transgene array at its site of integration. There were also strain-specific effects on transgene expression that appear to be independent of the inherent variegation. The transgene, compared to endogenous milk protein genes, is specifically susceptible to inbreeding depression. Outcrossing restored transgene expression levels to that of the parental population; thus suppression was not inherited. Finally, no generation-dependent decrease in mean expression levels was observed in the parental population. Thus, although the BLG/7 transgene is expressed in a variegated manner, there was no generation-associated accumulated silencing of transgene expression. PMID- 11901127 TI - Linkage disequilibrium in domestic sheep. AB - The last decade has seen a dramatic increase in the number of livestock QTL mapping studies. The next challenge awaiting livestock geneticists is to determine the actual genes responsible for variation of economically important traits. With the advent of high density single nucleotide polymorphism (SNP) maps, it may be possible to fine map genes by exploiting linkage disequilibrium between genes of interest and adjacent markers. However, the extent of linkage disequilibrium (LD) is generally unknown for livestock populations. In this article microsatellite genotype data are used to assess the extent of LD in two populations of domestic sheep. High levels of LD were found to extend for tens of centimorgans and declined as a function of marker distance. However, LD was also frequently observed between unlinked markers. The prospects for LD mapping in livestock appear encouraging provided that type I error can be minimized. Properties of the multiallelic LD coefficient D' were also explored. D' was found to be significantly related to marker heterozygosity, although the relationship did not appear to unduly influence the overall conclusions. Of potentially greater concern was the observation that D' may be skewed when rare alleles are present. It is recommended that the statistical significance of LD is used in conjunction with coefficients such as D' to determine the true extent of LD. PMID- 11901128 TI - Crossover interference in the mouse. AB - We present an analysis of crossover interference in the mouse genome, on the basis of high-density genotype data from two reciprocal interspecific backcrosses, comprising 188 meioses. Overwhelming evidence was found for strong positive crossover interference with average strength greater than that implied by the Carter-Falconer map function. There was some evidence for interchromosomal variation in the level of interference, with smaller chromosomes exhibiting stronger interference. We further compared the observed numbers of crossovers to previous cytological observations on the numbers of chiasmata and evaluated evidence for the obligate chiasma hypothesis. PMID- 11901129 TI - Quantitative trait loci for inflorescence development in Arabidopsis thaliana. AB - Variation in inflorescence development patterns is a central factor in the evolutionary ecology of plants. The genetic architectures of 13 traits associated with inflorescence developmental timing, architecture, rosette morphology, and fitness were investigated in Arabidopsis thaliana, a model plant system. There is substantial naturally occurring genetic variation for inflorescence development traits, with broad sense heritabilities computed from 21 Arabidopsis ecotypes ranging from 0.134 to 0.772. Genetic correlations are significant for most (64/78) pairs of traits, suggesting either pleiotropy or tight linkage among loci. Quantitative trait locus (QTL) mapping indicates 47 and 63 QTL for inflorescence developmental traits in Ler x Col and Cvi x Ler recombinant inbred mapping populations, respectively. Several QTL associated with different developmental traits map to the same Arabidopsis chromosomal regions, in agreement with the strong genetic correlations observed. Epistasis among QTL was observed only in the Cvi x Ler population, and only between regions on chromosomes 1 and 5. Examination of the completed Arabidopsis genome sequence in three QTL regions revealed between 375 and 783 genes per region. Previously identified flowering time, inflorescence architecture, floral meristem identity, and hormone signaling genes represent some of the many candidate genes in these regions. PMID- 11901130 TI - The colinearity of the Sh2/A1 orthologous region in rice, sorghum and maize is interrupted and accompanied by genome expansion in the triticeae. AB - The Sh2/A1 orthologous region of maize, rice, and sorghum contains five genes in the order Sh2, X1, X2, and two A1 homologs in tandem duplication. The Sh2 and A1 homologs are separated by approximately 20 kb in rice and sorghum and by approximately 140 kb in maize. We analyzed the fate of the Sh2/A1 region in large genome species of the Triticeae (wheat, barley, and rye). In the Triticeae, synteny in the Sh2/A1 region was interrupted by a break between the X1 and X2 genes. The A1 and X2 genes remained colinear in homeologous chromosomes as in other grasses. The Sh2 and X1 orthologs also remained colinear but were translocated to a nonhomeologous chromosome. Gene X1 was duplicated on two nonhomeologous chromosomes, and surprisingly, a paralog shared homology much higher than that of the orthologous copy to the X1 gene of other grasses. No tandem duplication of A1 homologs was detected but duplication of A1 on a nonhomeologous barley chromosome 6H was observed. Intergenic distances expanded greatly in wheat compared to rice. Wheat and barley diverged from each other 12 million years ago and both show similar changes in the Sh2/A1 region, suggesting that the break in colinearity as well as X1 duplications and genome expansion occurred in a common ancestor of the Triticeae species. PMID- 11901131 TI - Genetic analyses of endoreduplication in Zea mays endosperm: evidence of sporophytic and zygotic maternal control. AB - Flow cytometry was used to assess the variability of endoreduplication in endosperms of maize inbred lines. Little variation was found between midwestern dent types, and high levels of endoreduplication were observed in popcorns. Endoreduplication is different between inbred lines by 13-18 days after pollination, and flow cytometric analysis of ploidy level was feasible until 20 DAP. To study the genetic regulation of endoreduplication, four inbreds were crossed to B73 and developing endosperms from both parental, reciprocal F(1), and backcross generations were subjected to flow cytometric analysis. Three measurements of endoreduplication were calculated from these data and analyzed as quantitative genetic traits. Multiple models of trait inheritance were considered including triploid, diploid, sporophytic maternal, and maternal and paternal zygotic nuclear inheritance. Maternal zygotic effects, often considered a form of parental imprinting, and maternal sporophytic effects were detected. To test the feasibility of introgressing a high endoreduplication phenotype into a midwestern dent inbred line, a backcross population was generated from B73 x Sg18. Parental and progeny endoreduplication levels were compared and heritabilities assessed. The heritabilities calculated from these data generally agree with the values calculated in the larger crossing experiments. PMID- 11901132 TI - The signature of positive selection at randomly chosen loci. AB - In Drosophila and humans, there are accumulating examples of loci with a significant excess of high-frequency-derived alleles or high levels of linkage disequilibrium, relative to a neutral model of a random-mating population of constant size. These are features expected after a recent selective sweep. Their prevalence suggests that positive directional selection may be widespread in both species. However, as I show here, these features do not persist long after the sweep ends: The high-frequency alleles drift to fixation and no longer contribute to polymorphism, while linkage disequilibrium is broken down by recombination. As a result, loci chosen without independent evidence of recent selection are not expected to exhibit either of these features, even if they have been affected by numerous sweeps in their genealogical history. How then can we explain the patterns in the data? One possibility is population structure, with unequal sampling from different subpopulations. Alternatively, positive selection may not operate as is commonly modeled. In particular, the rate of fixation of advantageous mutations may have increased in the recent past. PMID- 11901133 TI - Selection, load and inbreeding depression in a large metapopulation. AB - The subdivision of a species into local populations causes its response to selection to change, even if selection is uniform across space. Population structure increases the frequency of homozygotes and therefore makes selection on homozygous effects more effective. However, population subdivision can increase the probability of competition among relatives, which may reduce the efficacy of selection. As a result, the response to selection can be either increased or decreased in a subdivided population relative to an undivided one, depending on the dominance coefficient F(ST) and whether selection is hard or soft. Realistic levels of population structure tend to reduce the mean frequency of deleterious alleles. The mutation load tends to be decreased in a subdivided population for recessive alleles, as does the expected inbreeding depression. The magnitude of the effects of population subdivision tends to be greatest in species with hard selection rather than soft selection. Population structure can play an important role in determining the mean fitness of populations at equilibrium between mutation and selection. PMID- 11901134 TI - An estimator for pairwise relatedness using molecular markers. AB - I propose a new estimator for jointly estimating two-gene and four-gene coefficients of relatedness between individuals from an outbreeding population with data on codominant genetic markers and compare it, by Monte Carlo simulations, to previous ones in precision and accuracy for different distributions of population allele frequencies, numbers of alleles per locus, actual relationships, sample sizes, and proportions of relatives included in samples. In contrast to several previous estimators, the new estimator is well behaved and applies to any number of alleles per locus and any allele frequency distribution. The estimates for two- and four-gene coefficients of relatedness from the new estimator are unbiased irrespective of the sample size and have sampling variances decreasing consistently with an increasing number of alleles per locus to the minimum asymptotic values determined by the variation in identity-by-descent among loci per se, regardless of the actual relationship. The new estimator is also robust for small sample sizes and for unknown relatives being included in samples for estimating allele frequencies. Compared to previous estimators, the new one is generally advantageous, especially for highly polymorphic loci and/or small sample sizes. PMID- 11901135 TI - A model-based method for identifying species hybrids using multilocus genetic data. AB - We present a statistical method for identifying species hybrids using data on multiple, unlinked markers. The method does not require that allele frequencies be known in the parental species nor that separate, pure samples of the parental species be available. The method is suitable for both markers with fixed allelic differences between the species and markers without fixed differences. The probability model used is one in which parentals and various classes of hybrids (F(1)'s, F(2)'s, and various backcrosses) form a mixture from which the sample is drawn. Using the framework of Bayesian model-based clustering allows us to compute, by Markov chain Monte Carlo, the posterior probability that each individual belongs to each of the distinct hybrid classes. We demonstrate the method on allozyme data from two species of hybridizing trout, as well as on two simulated data sets. PMID- 11901136 TI - A coalescent-based method for detecting and estimating recombination from gene sequences. AB - Determining the amount of recombination in the genealogical history of a sample of genes is important to both evolutionary biology and medical population genetics. However, recurrent mutation can produce patterns of genetic diversity similar to those generated by recombination and can bias estimates of the population recombination rate. Hudson 2001 has suggested an approximate likelihood method based on coalescent theory to estimate the population recombination rate, 4N(e)r, under an infinite-sites model of sequence evolution. Here we extend the method to the estimation of the recombination rate in genomes, such as those of many viruses and bacteria, where the rate of recurrent mutation is high. In addition, we develop a powerful permutation-based method for detecting recombination that is both more powerful than other permutation-based methods and robust to misspecification of the model of sequence evolution. We apply the method to sequence data from viruses, bacteria, and human mitochondrial DNA. The extremely high level of recombination detected in both HIV1 and HIV2 sequences demonstrates that recombination cannot be ignored in the analysis of viral population genetic data. PMID- 11901137 TI - Modeling epistasis of quantitative trait loci using Cockerham's model. AB - We use the orthogonal contrast scales proposed by Cockerham to construct a genetic model, called Cockerham's model, for studying epistasis between genes. The properties of Cockerham's model in modeling and mapping epistatic genes under linkage equilibrium and disequilibrium are investigated and discussed. Because of its orthogonal property, Cockerham's model has several advantages in partitioning genetic variance into components, interpreting and estimating gene effects, and application to quantitative trait loci (QTL) mapping when compared to other models, and thus it can facilitate the study of epistasis between genes and be readily used in QTL mapping. The issues of QTL mapping with epistasis are also addressed. Real and simulated examples are used to illustrate Cockerham's model, compare different models, and map for epistatic QTL. Finally, we extend Cockerham's model to multiple loci and discuss its applications to QTL mapping. PMID- 11901143 TI - Structure of the globular tail of nuclear lamin. AB - The nuclear lamins form a two-dimensional matrix that provides integrity to the cell nucleus and participates in nuclear activities. Mutations in the region of human LMNA encoding the carboxyl-terminal tail Lamin A/C are associated with forms of muscular dystrophy and familial partial lipodystrophy (FPLD). To help discriminate tissue-specific phenotypes, we have solved at 1.4-A resolution the three-dimensional crystal structure of the lamin A/C globular tail. The domain adopts a novel, all beta immunoglobulin-like fold. FPLD-associated mutations cluster within a small surface, whereas muscular dystrophy-associated mutations are distributed throughout the protein core and on its surface. These findings distinguish myopathy- and lipodystrophy-associated mutations and provide a structural framework for further testing hypotheses concerning lamin function. PMID- 11901144 TI - Identification, characterization, and localization of a novel kidney polycystin-1 polycystin-2 complex. AB - The functions of the two proteins defective in autosomal dominant polycystic kidney disease, polycystin-1 and polycystin-2, have not been fully clarified, but it has been hypothesized that they may heterodimerize to form a "polycystin complex" involved in cell adhesion. In this paper, we demonstrate for the first time the existence of a native polycystin complex in mouse kidney tubular cells transgenic for PKD1, non-transgenic kidney cells, and normal adult human kidney. Polycystin-1 is heavily N-glycosylated, and several glycosylated forms of polycystin-1 differing in their sensitivity to endoglycosidase H (Endo H) were found; in contrast, native polycystin-2 was fully Endo H-sensitive. Using highly specific antibodies to both proteins, we show that polycystin-2 associates selectively with two species of full-length polycystin-1, one Endo H-sensitive and the other Endo H-resistant; importantly, the latter could be further enriched in plasma membrane fractions and co-immunoprecipitated with polycystin-2. Finally, a subpopulation of this complex co-localized to the lateral cell borders of PKD1 transgenic kidney cells. These results demonstrate that polycystin-1 and polycystin-2 interact in vivo to form a stable heterodimeric complex and suggest that disruption of this complex is likely to be of primary relevance to the pathogenesis of cyst formation in autosomal dominant polycystic kidney disease. PMID- 11901145 TI - alpha-Thrombin induces rapid and sustained Akt phosphorylation by beta-arrestin1 dependent and -independent mechanisms, and only the sustained Akt phosphorylation is essential for G1 phase progression. AB - In Chinese hamster embryonic fibroblasts (IIC9 cells) alpha-thrombin activates the MAPK(ERK) and phosphatidylinositol 3-OH-kinase (PI 3-kinase)/Akt pathways, and both are essential for progression through the G(1) phase of the cell cycle. We investigated in IIC9 cells, the role of beta-arrestin1 in alpha-thrombin signaling to these pathways. alpha-Thrombin stimulates rapid and sustained PI 3 kinase and Akt activities. Expression of a dominant negative beta-arrestin1 (beta arrestin1(V53D)) inhibits rapid but not sustained PI 3-kinase and Akt activities. Surprisingly, expression of beta-arrestin1(V53D) does not block activation of the MAPK(ERK) pathway. PI 3-kinase and Akt activities are also inhibited by expression of a beta-arrestin1 mutant, which impairs binding to c-Src (beta arrestin1(P91G-P121E)), indicating the involvement of c-Src in the rapid stimulation of the PI 3-kinase/Akt pathway. Consistent with these results, PP1, a selective inhibitor of c-Src family kinases, prevents alpha-thrombin-stimulated Akt phosphorylation. Expression of beta- arrestin1(V53D) does not prevent G(1) progression, as its expression has no effect on [(3)H]thymidine incorporation into DNA. In agreement with the ineffectiveness of beta-arrestin1(V53D) to block G(1) progression, cyclin D1 protein amounts and CDK4-cyclin D1 activity is unaffected by expression of beta-arrestin1(V53D). Thus in IIC9 cells, alpha thrombin activates rapid beta-arrestin1-dependent and sustained beta-arrestin1 independent Akt activity, suggesting that two mechanisms are involved. Furthermore, although blocking the beta-arrestin1-independent PI 3-kinase/Akt pathway prevents G(1) progression, inhibition of the beta-arrestin1-dependent pathway does not, indicating different roles for the rapid and sustained activities. PMID- 11901146 TI - Regulation of ATP-binding cassette sterol transporters ABCG5 and ABCG8 by the liver X receptors alpha and beta. AB - Mutations in the ATP-binding cassette (ABC) transporters ABCG5 and ABCG8 have recently been shown to cause the autosomal recessive disorder sitosterolemia. Here we demonstrate that the ABCG5 and ABCG8 genes are direct targets of the oxysterol receptors liver X receptor (LXR) alpha and LXRbeta. Diets containing high cholesterol markedly increased the expression of ABCG5/G8 mRNA in mouse liver and intestine. This increase was also observed using synthetic ligands of LXR and its heterodimeric partner, the retinoid X receptor. In situ hybridization analyses of tissues from LXR agonist-treated mice revealed that ABCG5/G8 mRNA is located in hepatocytes and enterocytes and is increased upon LXR activation. In addition, expression of the LXR target gene ABCA1, previously implicated in the control of cholesterol absorption, was also dramatically up-regulated in jejunal enterocytes upon exposure to LXR agonists. These changes in ABC transporter gene expression were not observed in mice lacking LXRs. Furthermore, in the rat hepatoma cell line FTO2B, LXR-dependent transcription of the ABCG5/G8 genes was cycloheximide-resistant, indicating that these genes are directly regulated by LXRs. The addition of ABCG5 and ABCG8 to the growing list of LXR target genes further supports the notion that LXRs serve as sterol sensors to coordinately regulate sterol catabolism, storage, efflux, and elimination. PMID- 11901147 TI - Structural analysis of the extracellular domain of vaccinia virus envelope protein, A27L, by NMR and CD spectroscopy. AB - This study presents the molecular structure of the extracellular domain of vaccinia virus envelope protein, A27L, determined by NMR and CD spectroscopy. A recombinant protein, eA27L-aa, containing this domain in which cysteines 71 and 72 were replaced with alanine, was constructed to prevent self-assembly due to intermolecular disulfide bonds between these two cysteines. The soluble eA27L-aa protein forms an oligomer resembling that of A27L on vaccinia virions. Heteronuclear correlation NMR spectroscopy was carried out on eA27L-aa in the presence or absence of urea to determine backbone resonance assignments. Chemical shift index (CSI) propensity analysis showed that eA27L-aa has two distinct structural domains, a relatively flexible 22-amino acid random coil in the N terminal region and a fairly rigid alpha-helix structure in the remainder of the structure. Binding interaction studies using isothermal titration calorimetry suggest that a 12-amino acid lysine/arginine-rich segment in the N-terminal region is responsible for glycosaminoglycan binding. The rigid alpha-helix portion of eA27L-aa is probably involved in the intrinsic self-assembly, and CSI propensity analysis suggests that region N37-E49, with a residual alpha-helix tendency, is probably the self-assembly core. Self-assembly was ascribed to three hydrophobic leucine residues (Leu(41), Leu(45), and Leu(48)) in this segment. The folding mechanism of eA27L-aa was analyzed by CD spectroscopy, which revealed a two-step transition with a Gibbs free energy of 2.5 kcal/mol in the absence of urea. Based on these NMR and CD studies, a residue-specific molecular model of the extracellular domain of A27L is proposed. These studies on the molecular structure of eA27L-aa will help in understanding how vaccinia virus enters cells. PMID- 11901148 TI - Identification of calmodulin isoform-specific binding peptides from a phage displayed random 22-mer peptide library. AB - Plants express numerous calmodulin (CaM) isoforms that exhibit differential activation or inhibition of CaM-dependent enzymes in vitro; however, their specificities toward target enzyme/protein binding are uncertain. A random peptide library displaying a 22-mer peptide on a bacteriophage surface was constructed to screen peptides that specifically bind to plant CaM isoforms (soybean calmodulin (ScaM)-1 and SCaM-4 were used in this study) in a Ca2+ dependent manner. The deduced amino acid sequence analyses of the respective 80 phage clones that were independently isolated via affinity panning revealed that SCaM isoforms require distinct amino acid sequences for optimal binding. SCaM-1 binding peptides conform to a 1-5-10 ((FILVW)XXX(FILV) XXXX(FILVW)) motif (where X denotes any amino acid), whereas SCaM-4-binding peptide sequences conform to a 1-8-14 ((FILVW)XXXXXX(FAILVW)XXXXX(FILVW)) motif. These motifs are classified based on the positions of conserved hydrophobic residues. To examine their binding properties further, two representative peptides from each of the SCaM isoform-binding sequences were synthesized and analyzed via gel mobility shift assays, Trp fluorescent spectra analyses, and phosphodiesterase competitive inhibition experiments. The results of these studies suggest that SCaM isoforms possess different binding sequences for optimal target interaction, which therefore may provide a molecular basis for CaM isoform-specific function in plants. Furthermore, the isolated peptide sequences may serve not only as useful CaM-binding sequence references but also as potential reagents for studying CaM isoform-specific function in vivo. PMID- 11901149 TI - Primer unblocking and rescue of DNA synthesis by azidothymidine (AZT)-resistant HIV-1 reverse transcriptase: comparison between initiation and elongation of reverse transcription and between (-) and (+) strand DNA synthesis. AB - Azidothymidine (AZT) is a widely used inhibitor of type 1 human immunodeficiency virus reverse transcriptase (RT) that acts as chain terminator. Upon treatment, mutations conferring AZT resistance to RT are gradually selected. It has been shown that resistant RT is able to unblock the AZT-terminated primer by an ATP dependent mechanism. However, this resistance mechanism has only been demonstrated for DNA-dependent DNA elongation. Here, we compared the AZT resistance of mutant RT during DNA elongation on DNA and RNA templates. We showed that, during DNA elongation, primer unblocking and rescue of DNA synthesis take place with similar rate constants on DNA and RNA templates. However, the fraction of a primer eventually repaired during RNA-dependent DNA synthesis is 2x lower compared with that of DNA-dependent synthesis, leading to reduced resistance. We also compared the initiation of reverse transcription, which uses tRNA(3)(Lys) as a primer and displays characteristic kinetic features, and the subsequent RNA dependent elongation. Unlike during elongation, resistant RT was unable to unblock the AZT-terminated primer during initiation of (-) DNA strand synthesis. Our results demonstrate that the efficiency of primer unblocking conferred by the AZT resistance mutations greatly vary during the different steps of the provirus synthesis. These results also suggest that inhibitors specifically targeting the initiation of reverse transcription might prove to be advantageous, as compared with elongation inhibitors. PMID- 11901150 TI - Three-dimensional modeling of thrombin-fibrinogen interaction. AB - Three-dimensional models of thrombin complexed with large fragments of the fibrinogen Aalpha and Bbeta chains are presented. The models are consistent with the results of recent mutagenesis studies of thrombin and with the information available on naturally occurring fibrinogen mutants. Thrombin recognizes fibrinogen with an extended binding surface, key elements of which are Tyr(76) in exosite I, located about 20 A away from the active site, and the aryl binding site located in close proximity to the catalytic triad. A highly conserved aromatic-Pro-aromatic triplet motif is identified in the primed site region of fibrinogen and other natural substrates of thrombin. The role of this triplet, based on the three-dimensional models, is to correctly orient the substrate for optimal bridge binding to exosite I and the active site. The three-dimensional models suggest a possible pattern of recognition by thrombin that applies generally to other natural substrates. PMID- 11901151 TI - Cyclooxygenase-2 is overexpressed in HER-2/neu-positive breast cancer: evidence for involvement of AP-1 and PEA3. AB - Markedly increased levels of cyclooxygenase-2 (COX-2) mRNA, protein, and prostaglandin E(2) synthesis were detected in HER-2/neu-transformed human mammary epithelial cells (184B5/HER) compared with its nontransformed partner cell line (184B5). HER-2/neu stimulated COX-2 transcription via the Ras --> Raf --> MAPK pathway. The inductive effects of HER-2/neu were mediated, in part, by enhanced binding of AP-1 (c-Jun, c-Fos, and ATF-2) to the cyclic AMP-response element ( 59/-53) of the COX-2 promoter. The potential contribution of the transcription factor PEA3 was also investigated. Elevated levels of PEA3 were detected in 184B5/HER cells. A PEA3 site (-75/-72) was identified juxtaposed to the cyclic AMP-response element. HER-2/neu-mediated activation of the COX-2 promoter was blocked by mutagenizing the PEA3 site or overexpressing antisense to PEA3. To determine whether HER-2/neu status was also a determinant of COX-2 expression in vivo, we compared levels of COX-2 protein in HER-2/neu-positive and -negative human breast cancers. Increased amounts of COX-2 were detected in HER-2/neu positive tumors. Taken together, these results suggest that closely spaced PEA3 and cyclic AMP-response elements are required for HER-2/neu-mediated induction of COX-2 transcription. The clear relationship between HER-2/neu status and COX-2 expression in human breast tumors suggests that this mechanism is likely to be operative in vivo. PMID- 11901153 TI - Protein kinase C-beta II Is an apoptotic lamin kinase in polyomavirus transformed, etoposide-treated pyF111 rat fibroblasts. AB - The role of protein kinase C-beta(II) (PKC-beta(II)) in etoposide (VP-16)-induced apoptosis was studied using polyomavirus-transformed pyF111 rat fibroblasts in which PKC-beta(II) specific activity in the nuclear membrane (NM) doubled and the enzyme was cleaved into catalytic fragments. No PKC-beta(II) complexes with lamin B1 and/or active caspases were immunoprecipitable from the NM of proliferating untreated cells, but large complexes of PKC-beta(II) holoprotein and its catalytic fragments with lamin B1, active caspase-3 and -6, and inactive phospho CDK-1, but not PKC-beta(I) or PKC-delta, could be immunoprecipitated from the NM of VP-16-treated cells, suggesting that PKC-beta(II) is an apoptotic lamin kinase. By 30 min after normal nuclei were mixed with cytoplasms from VP-16 treated, but not untreated, cells, PKC-beta(II) holoprotein had moved from the apoptotic cytoplasm to the normal NM, and lamin B1 was phosphorylated before cleavage by caspase-6. Lamin B1 phosphorylation was partly reduced, but its cleavage was completely prevented, despite the presence of active caspase-6, by adding a selective PKC-betas inhibitor, hispidin, to the apoptotic cytoplasms. Thus, a PKC-beta(II) response to VP-16 seems necessary for lamin B1 cleavage by caspase-6 and nuclear lamina dissolution in apoptosing pyF111 fibroblasts. The possibility of PKC-beta(II) being an apoptotic lamin kinase in these cells was further suggested by lamin B1-bound PKC-delta being inactive or only slightly active and by PKC-alpha not combining with the lamin. PMID- 11901152 TI - Clustered charged amino acids of human adenosine deaminase comprise a functional epitope for binding the adenosine deaminase complexing protein CD26/dipeptidyl peptidase IV. AB - Human adenosine deaminase (ADA) occurs as a 41-kDa soluble monomer in all cells. On epithelia and lymphoid cells of humans, but not mice, ADA also occurs bound to the membrane glycoprotein CD26/dipeptidyl peptidase IV. This "ecto-ADA" has been postulated to regulate extracellular Ado levels, and also the function of CD26 as a co-stimulator of activated T cells. The CD26-binding site of human ADA has been localized by homolog scanning to the peripheral alpha2-helix (amino acids 126 143). Among the 5 non-conserved residues within this segment, Arg-142 in human and Gln-142 in mouse ADA largely determined the capacity for stable binding to CD26 (Richard, E., Arredondo-Vega, F. X., Santisteban, I., Kelly, S. J., Patel, D. D., and Hershfield, M. S. (2000) J. Exp. Med. 192, 1223-1235). We have now mutagenized conserved alpha2-helix residues in human and mouse ADA and used surface plasmon resonance to evaluate binding kinetics to immobilized rabbit CD26. In addition to Arg-142, we found that Glu-139 and Asp-143 of human ADA are also important for CD26 binding. Mutating these residues to alanine increased dissociation rates 6-11-fold and the apparent dissociation constant K(D) for wild type human ADA from 17 to 112-160 nm, changing binding free energy by 1.1-1.3 kcal/mol. This cluster of 3 charged residues appears to be a "functional epitope" that accounts for about half of the difference between human and mouse ADA in free energy of binding to CD26. PMID- 11901154 TI - Spectral properties of bacterial nitric-oxide reductase: resolution of pH dependent forms of the active site heme b3. AB - Bacterial nitric-oxide reductase catalyzes the two electron reduction of nitric oxide to nitrous oxide. In the oxidized form the active site non-heme Fe(B) and high spin heme b(3) are mu-oxo bridged. The heme b(3) has a ligand-to-metal charge transfer band centered at 595 nm, which is insensitive to pH over the range of 6.0-8.5. Partial reduction of nitric-oxide reductase yields a three electron-reduced state where only the heme b(3) remains oxidized. This results in a shift of the heme b(3) charge transfer band lambda(max) to longer wavelengths. At pH 6.0 the charge transfer band lambda(max) is 605 nm, whereas at pH 8.5 it is 635 nm. At pH 6.5 and 7.5 the nitric-oxide reductase ferric heme b(3) population is a mixture of both 605- and 635-nm forms. Magnetic circular dichroism spectroscopy suggests that at all pH values examined the proximal ligand to the ferric heme b(3) in the three electron-reduced form is histidine. At pH 8.5 the distal ligand is hydroxide, whereas at pH 6.0, when the enzyme is most active, it is water. PMID- 11901155 TI - alpha 2-HS glycoprotein/fetuin, a transforming growth factor-beta/bone morphogenetic protein antagonist, regulates postnatal bone growth and remodeling. AB - Soluble transforming growth factor-beta (TGF-beta)/bone morphogenetic protein (BMP)-binding proteins are widely distributed in mammalian tissues and control cytokine access to membrane signaling receptors. The serum and bone-resident glycoprotein alpha2-HS-glycoprotein/fetuin (ASHG) binds to TGF-beta/BMP cytokines and blocks TGF-beta1 binding to cell surface receptors. Therefore, we examined bone growth and remodeling phenotypes in ASHG-deficient mice. The skeletal structure of Ahsg(-/-) mice appeared normal at birth, but abnormalities were observed in adult Ahsg(-/-) mice. Maturation of growth plate chondrocytes was impaired, and femurs lengthened more slowly between 3 and 18 months of age in Ahsg(-/-) mice. However, bone formation was increased in Ahsg(-/-) mice as indicated by greater cortical thickness, accelerated trabecular bone remodeling, and increased osteoblast numbers on bone surfaces. The normal age-related increase in cortical thickness and bone mineral density was accelerated in Ahsg( /-) mice and was associated with increased energy required to fracture. Bone formation in response to implanted BMP cytokine extended further from the implant in Ahsg(-/-) compared with Ahsg(+/+) mice, confirming the interaction between ASHG and TGF-beta/BMP cytokines in vivo. Our results demonstrate that ASHG blocks TGF-beta-dependent signaling in osteoblastic cells, and mice lacking ASHG display growth plate defects, increased bone formation with age, and enhanced cytokine dependent osteogenesis. PMID- 11901157 TI - Targeted disruption of the GAS41 gene encoding a putative transcription factor indicates that GAS41 is essential for cell viability. AB - The glioma-amplified sequence (GAS) 41 protein has been proposed to be a transcription factor. To investigate its functional role in vivo, we attempted to knock out the GAS41 gene by targeted disruption in the chicken pre-lymphoid cell line DT40. Heterozygous GAS41+/- cell lines generated by the first round of homologous recombination express approximately half the normal level of GAS41 mRNA. However, a homozygous GAS41-/- cell line with both GAS41 alleles disrupted was not obtained following the second round of transfection, indicating that the GAS41 gene is essential for cell viability. Indeed, homozygous GAS41-/- cell lines with two disrupted GAS41 alleles can be generated following substitution of the endogenous gene by stable integration of GAS41 cDNA controlled by a tetracycline-regulated CMV promoter. Inactivation of this promoter by tetracycline withdrawal results in rapid depletion of GAS41, causing a significant decrease in RNA synthesis and subsequently cell death. Thus, our results indicate that GAS41 is required for RNA transcription. PMID- 11901156 TI - Functional reconstitution of insulin receptor binding site from non-binding receptor fragments. AB - We have previously shown that a minimized insulin receptor (IR) consisting of the first 468 amino acids of the insulin receptor fused to 16 amino acids from the C terminus of the alpha-subunit (CT domain) bound insulin with nanomolar affinity (Kristensen, C., Wiberg, F. C., Schaffer, L., and Andersen, A. S. (1998) J. Biol. Chem. 273, 17780-17786). In the present study, we show that a smaller construct that has the first 308 residues fused to the CT domain also binds insulin. Insulin receptor fragments consisting of the first 468 or 308 residues did not bind insulin. However, when these fragments were mixed with a synthetic peptide corresponding to the CT domain, insulin binding was detectable. At concentrations of 10 microm CT peptide, insulin binding was fully reconstituted yielding apparent affinities of 9-11 nm. To further investigate the minimum requirement for the length of the N terminus of IR, we tested smaller receptor fragments for insulin binding in the presence of the CT peptide and found that a fragment consisting of the first 255 amino acids of IR was able to fully reconstitute the insulin binding site, yielding an apparent affinity of 11 +/- 4 nm for insulin. PMID- 11901158 TI - Phosphorylation of threonine 68 promotes oligomerization and autophosphorylation of the Chk2 protein kinase via the forkhead-associated domain. AB - Phosphorylation of Thr-68 by the ataxia telangiectasia-mutated is necessary for efficient activation of Chk2 when cells are exposed to ionizing radiation. By an unknown mechanism, this initial event promotes additional autophosphorylation events including modifications of Thr-383 and Thr-387, two amino acid residues located within the activation loop segment within the Chk2 catalytic domain. Chk2 and related kinases possess one or more Forkhead-associated (FHA) domains that are phosphopeptide-binding modules believed to be crucial for their checkpoint control activities. We show that the Chk2 FHA domain is dispensable for Thr-68 phosphorylation but necessary for efficient autophosphorylation in response to ionizing radiation. Phosphorylation of Thr-68 promotes oligomerization of Chk2 by serving as a specific ligand for the FHA domain of another Chk2 molecule. In addition, Chk2 phosphorylates its own FHA domain, and this modification reduces its affinity for Thr-68-phosphorylated Chk2. Thus, activation of Chk2 in irradiated cells may occur through oligomerization of Chk2 via binding of the Thr 68-phosphorylated region of one Chk2 to the FHA domain of another. Oligomerization of Chk2 may therefore increase the efficiency of trans autophosphorylation resulting in the release of active Chk2 monomers that proceed to enforce checkpoint control in irradiated cells. PMID- 11901159 TI - Human Herpesvirus 6 immediate-early 1 protein is a sumoylated nuclear phosphoprotein colocalizing with promyelocytic leukemia protein-associated nuclear bodies. AB - Immediate-early (IE) proteins are the first proteins expressed following viral entry and play a crucial role in the initiation of infection. We report the cloning and characterization of a full-length IE1 transcript and protein (IE1B) from human herpesvirus 6 (HHV-6) variant B. The IE1B transcript consists of five exons (3720 nucleotides), three of which are coding for the IE1 protein. The 1078 amino acid-long IE1B protein is 62% identical and 75% similar to the 941-amino acid IE1 from HHV-6 variant A. IE1B protein can be detected at 4 h post-infection (P.I.), and it is distributed as small intranuclear structures. The maximal number of IE1 bodies ( approximately 10-12/nucleus) is detected at 12 h P.I. after which the IE1 bodies condense into 1-3 larger entities by 24-48 h P.I. During infection the IE1B protein is phosphorylated on serine and threonine residues. IE1B undergoes further post-translational modification with its conjugation to the small ubiquitin-like modifier (SUMO-1) peptide. IE1B colocalizes with SUMO-1 and promyelocytic leukemia nuclear bodies during infection as well as in transfection experiments. Finally, IE1 from variant B is a weaker transactivator than IE1 from variant A, when assayed using heterologous promoters. Overall, the characterization of the HHV-6 IE1B protein presented highlights the similarity and divergence between IE1 from both variants and provides useful information pertaining to the early phase of infection. PMID- 11901160 TI - Eicosanoid regulation of vascular endothelial growth factor expression and angiogenesis in microvessel endothelial cells. AB - 12(R)-Hydroxy-5,8,14-eicosatrienoic acid (HETrE) is a potent inflammatory and angiogenic eicosanoid in ocular and dermal tissues. Previous studies suggested that 12(R)-HETrE activates microvessel endothelial cells via a high affinity binding site; however, the cellular mechanisms underlying 12(R)-HETrE angiogenic activity are unexplored. Because the synthesis of 12(R)-HETrE is induced in response to hypoxic injury, we examined its interactions with vascular endothelial growth factor (VEGF) in rabbit limbal microvessel endothelial cells. Addition of 12(R)-HETrE (0.1 nm) to the cells increased VEGF mRNA levels with maximum 5-fold increase at 45 min. The increase in VEGF mRNA was followed by an increase in immunoreactive VEGF protein. 12(R)-HETrE (0.1 nm) rapidly activated the extracellular signal-regulated kinases (ERKs) ERK1 and ERK2. Moreover, preincubation of cells with PD98059, a selective inhibitor of MEK-1, inhibited 12(R)-HETrE-induced VEGF mRNA. Addition of VEGF antibody to cells grown in Matrigel-coated culture plates inhibited 12(R)-HETrE-induced capillary tube-like formation, suggesting that VEGF mediates, at least in part, the angiogenic response to 12(R)-HETrE. The results indicate that in microvessel endothelial cells, 12(R)-HETrE induces VEGF expression via activation of ERK1/2 and that VEGF mediates, at least in part, the angiogenic activity of 12(R)-HETrE. Given the fact that both VEGF and 12(R)-HETrE are produced in the cornea after hypoxic injury, their interaction may be an important determinant in the development of neovascularized tissues. PMID- 11901161 TI - Mechanisms regulating adipocyte expression of resistin. AB - Resistin, also known as Adipocyte Secreted Factor (ADSF) and Found in Inflammatory Zone 3 (FIZZ3), is a mouse protein with potential roles in insulin resistance and adipocyte differentiation. The resistin gene is expressed almost exclusively in adipocytes. Here we show that a proximal 264-base pair fragment of the mouse resistin promoter is sufficient for expression in adipocytes. Ectopic expression of the adipogenic transcription factor CCAAT/enhancer-binding protein (C/EBPalpha) was sufficient for expression in non-adipogenic cells. C/EBPalpha binds specifically to a site that is essential for expression of the resistin promoter. Chromatin immunoprecipitation studies of the endogenous gene demonstrated adipocyte-specific association of C/EBPalpha with the proximal resistin promoter in adipocytes but not preadipocytes. C/EBPalpha binding was associated with the recruitment of coactivators p300 and CREB-binding protein and a dramatic increase in histone acetylation in the vicinity of the resistin promoter. The antidiabetic thiazolidinedione (TZD) drug rosiglitazone reduced resistin expression with an ED(50) similar to its K(d) for binding to peroxisome proliferator activated receptor gamma (PPARgamma). Other TZD- and non-TZD PPARgamma ligands also down-regulated resistin expression. However, no functional PPARgamma binding site was found within 6.2 kb of the transcriptional start site, suggesting that if PPARgamma is involved, it is either acting at a long distance from the start site, in an intron, or indirectly. Nevertheless, rosiglitazone treatment selectively decreased histone acetylation at the resistin promoter without a change in occupation by C/EBPalpha, CREB-binding protein, or p300. Thus, adipocyte specificity of resistin gene expression is because of C/EBPalpha binding, leading to the recruitment of transcriptional coactivators and histone acetylation that is characteristic of an active chromatin environment. TZD reduces resistin gene expression at least in part by reducing histone acetylation associated with the binding of C/EBPalpha in mature adipocytes. PMID- 11901162 TI - Listeriolysin O: a genuine cytolysin optimized for an intracellular parasite. AB - Cholesterol-dependent cytolysins (CDCs)* are produced by a large number of pathogenic gram-positive bacteria. A member of this family, listeriolysin O (LLO), is produced by the intracellular pathogen Listeria monocytogenes. A unique feature of LLO is its low optimal pH activity (approximately 6) which permits escape of the bacterium from the phagosome into the host cell cytosol without damaging the plasma membrane of the infected cell. In a recent study (Glomski et al., 2002, this issue), Portnoy's group has addressed the molecular mechanism underlying the pH sensitivity of LLO. Unexpectedly, a single amino acid substitution in LLO L461T results in a molecule more active at neutral pH and promoting premature permeabilization of the infected cells, leading to attenuated virulence. This finding highlights how subtle changes in proteins can be exploited by bacterial pathogens to establish and maintain the integrity of their specific niches. PMID- 11901163 TI - Keratinocyte junctions and the epidermal barrier: how to make a skin-tight dress. AB - Although intercellular junctions are known to be the major regulators of permeability of simple epithelia, they had not been thought to be important in regulating the permeability of stratified mammalian epithelia. Furuse et al. (2002)(this issue) demonstrate that functional tight junctions may indeed be a necessary part of the permeability barrier of the skin. PMID- 11901165 TI - Cyclin-dependent kinases govern formation and maintenance of the nucleolus. AB - In higher eukaryotic cells, the nucleolus is a nuclear compartment assembled at the beginning of interphase, maintained during interphase, and disorganized during mitosis. Even if its structural organization appears to be undissociable from its function in ribosome biogenesis, the mechanisms that govern the formation and maintenance of the nucleolus are not elucidated. To determine if cell cycle regulators are implicated, we investigated the putative role of the cyclin-dependent kinases (CDKs) on ribosome biogenesis and nucleolar organization. Inhibition of CDK1-cyclin B during mitosis leads to resumption of rDNA transcription, but is not sufficient to induce proper processing of the pre rRNA and total relocalization of the processing machinery into rDNA transcription sites. Similarly, at the exit from mitosis, both translocation of the late processing machinery and pre-rRNA processing are impaired in a reversible manner by CDK inhibitors. Therefore, CDK activity seems indispensable for the building of functional nucleoli. Furthermore, inhibition of CDKs in interphasic cells also hampered proper pre-rRNA processing and induced a dramatic disorganization of the nucleolus. Thus, we propose that the mechanisms governing both formation and maintenance of functional nucleoli involve CDK activities and couple the cell cycle to ribosome biogenesis. PMID- 11901164 TI - CD46 is phosphorylated at tyrosine 354 upon infection of epithelial cells by Neisseria gonorrhoeae. AB - The Neisseria type IV pilus promotes bacterial adhesion to host cells. The pilus binds CD46, a complement-regulatory glycoprotein present on nucleated human cells (Kallstrom et al., 1997). CD46 mutants with truncated cytoplasmic tails fail to support bacterial adhesion (Kallstrom et al., 2001), suggesting that this region of the molecule also plays an important role in infection. Here, we report that infection of human epithelial cells by piliated Neisseria gonorrhoeae (GC) leads to rapid tyrosine phosphorylation of CD46. Studies with wild-type and mutant tail fusion constructs demonstrate that Src kinase phosphorylates tyrosine 354 in the Cyt2 isoform of the CD46 cytoplasmic tail. Consistent with these findings, infection studies show that PP2, a specific Src family kinase inhibitor, but not PP3, an inactive variant of this drug, reduces the ability of epithelial cells to support bacterial adhesion. Several lines of evidence point to the role of c-Yes, a member of the Src family of nonreceptor tyrosine kinases, in CD46 phosphorylation. GC infection causes c-Yes to aggregate in the host cell cortex beneath adherent bacteria, increases binding of c-Yes to CD46, and stimulates c Yes kinase activity. Finally, c-Yes immunoprecipitated from epithelial cells is able to phosphorylate the wild-type Cyt2 tail but not the mutant derivative in which tyrosine 354 has been substituted with alanine. We conclude that GC infection leads to rapid tyrosine phosphorylation of the CD46 Cyt2 tail and that the Src kinase c-Yes is involved in this reaction. Together, the findings reported here and elsewhere strongly suggest that pilus binding to CD46 is not a simple static process. Rather, they support a model in which pilus interaction with CD46 promotes signaling cascades important for Neisseria infectivity. PMID- 11901166 TI - Induction of secretory pathway components in yeast is associated with increased stability of their mRNA. AB - The overexpression of certain membrane proteins is accompanied by a striking proliferation of intracellular membranes. One of the best characterized inducers of membrane proliferation is the 180-kD mammalian ribosome receptor (p180), whose expression in yeast results in increases in levels of mRNAs encoding proteins that function in the secretory pathway, and an elevation in the cell's ability to secrete proteins. In this study we demonstrate that neither the unfolded protein response nor increased transcription accounts for membrane proliferation or the observed increase in secretory pathway mRNAs. Rather, p180-induced up-regulation of certain secretory pathway transcripts is due to a p180-mediated increase in the longevity of these mRNA species, as determined by measurements of transcriptional activity and specific mRNA turnover. Moreover, we show that the longevity of mRNA in general is substantially promoted through the process of its targeting to the membrane of the endoplasmic reticulum. With respect to the terminal differentiation of secretory tissues, results from this model system provide insights into how the expression of a single protein, p180, could result in substantial morphological and functional changes. PMID- 11901167 TI - Alpha-glucosidase I is required for cellulose biosynthesis and morphogenesis in Arabidopsis. AB - Novel mutations in the RSW1 and KNOPF genes were identified in a large-scale screen for mutations that affect cell expansion in early Arabidopsis embryos. Embryos from both types of mutants were radially swollen with greatly reduced levels of crystalline cellulose, the principal structural component of the cell wall. Because RSW1 was previously shown to encode a catalytic subunit of cellulose synthase, the similar morphology of knf and rsw1-2 embryos suggests that the radially swollen phenotype of knf mutants is largely due to their cellulose deficiency. Map-based cloning of the KNF gene and enzyme assays of knf embryos demonstrated that KNF encodes alpha-glucosidase I, the enzyme that catalyzes the first step in N-linked glycan processing. The strongly reduced cellulose content of knf mutants indicates that N-linked glycans are required for cellulose biosynthesis. Because cellulose synthase catalytic subunits do not appear to be N glycosylated, the N-glycan requirement apparently resides in other component(s) of the cellulose synthase machinery. Remarkably, cellular processes other than extracellular matrix biosynthesis and the formation of protein storage vacuoles appear unaffected in knf embryos. Thus in Arabidopsis cells, like yeast, N-glycan trimming is apparently required for the function of only a small subset of N-glycoproteins. PMID- 11901168 TI - The Listeria monocytogenes hemolysin has an acidic pH optimum to compartmentalize activity and prevent damage to infected host cells. AB - Listeria monocytogenes is a facultative intracellular bacterial pathogen that escapes from a phagosome and grows in the host cell cytosol. The pore-forming cholesterol-dependent cytolysin, listeriolysin O (LLO), mediates bacterial escape from vesicles and is approximately 10-fold more active at an acidic than neutral pH. By swapping dissimilar residues from a pH-insensitive orthologue, perfringolysin O (PFO), we identified leucine 461 as unique to pathogenic Listeria and responsible for the acidic pH optimum of LLO. Conversion of leucine 461 to the threonine present in PFO increased the hemolytic activity of LLO almost 10-fold at a neutral pH. L. monocytogenes synthesizing LLO L461T, expressed from its endogenous site on the bacterial chromosome, resulted in a 100 fold virulence defect in the mouse listeriosis model. These bacteria escaped from acidic phagosomes and initially grew normally in cells and spread cell to cell, but prematurely permeabilized the host membrane and killed the cell. These data show that the acidic pH optimum of LLO results from an adaptive mutation that acts to limit cytolytic activity to acidic vesicles and prevent damage in the host cytosol, a strategy also used by host cells to compartmentalize lysosomal hydrolases. PMID- 11901169 TI - A novel polymer of tubulin forms the conoid of Toxoplasma gondii. AB - Toxoplasma gondii is an obligatory intracellular parasite, an important human pathogen, and a convenient laboratory model for many other human and veterinary pathogens in the phylum Apicomplexa, such as Plasmodium, Eimeria, and Cryptosporidia. 22 subpellicular microtubules form a scaffold that defines the cell shape of T. gondii. Its cytoskeleton also includes an intricate apical structure consisting of the conoid, two intraconoid microtubules, and two polar rings. The conoid is a 380-nm diameter motile organelle, consisting of fibers wound into a spiral like a compressed spring. FRAP analysis of transgenic T. gondii expressing YFP-alpha-tubulin reveals that the conoid fibers are assembled by rapid incorporation of tubulin subunits during early, but not late, stages of cell division. Electron microscopic analysis shows that in the mature conoid, tubulin is arranged into a novel polymer form that is quite different from typical microtubules. PMID- 11901170 TI - Tau blocks traffic of organelles, neurofilaments, and APP vesicles in neurons and enhances oxidative stress. AB - We studied the effect of microtubule-associated tau protein on trafficking of vesicles and organelles in primary cortical neurons, retinal ganglion cells, and neuroblastoma cells. Tau inhibits kinesin-dependent transport of peroxisomes, neurofilaments, and Golgi-derived vesicles into neurites. Loss of peroxisomes makes cells vulnerable to oxidative stress and leads to degeneration. In particular, tau inhibits transport of amyloid precursor protein (APP) into axons and dendrites, causing its accumulation in the cell body. APP tagged with yellow fluorescent protein and transfected by adenovirus associates with vesicles moving rapidly forward in the axon (approximately 80%) and slowly back (approximately 20%). Both movements are strongly inhibited by cotransfection with fluorescently tagged tau (cyan fluorescent protein-tau) as seen by two-color confocal microscopy. The data suggests a linkage between tau and APP trafficking, which may be significant in Alzheimer's disease. PMID- 11901171 TI - Tropomyosin inhibits ADF/cofilin-dependent actin filament dynamics. AB - Tropomyosin binds to actin filaments and is implicated in stabilization of actin cytoskeleton. We examined biochemical and cell biological properties of Caenorhabditis elegans tropomyosin (CeTM) and obtained evidence that CeTM is antagonistic to ADF/cofilin-dependent actin filament dynamics. We purified CeTM, actin, and UNC-60B (a muscle-specific ADF/cofilin isoform), all of which are derived from C. elegans, and showed that CeTM and UNC-60B bound to F-actin in a mutually exclusive manner. CeTM inhibited UNC-60B-induced actin depolymerization and enhancement of actin polymerization. Within isolated native thin filaments, actin and CeTM were detected as major components, whereas UNC-60B was present at a trace amount. Purified UNC-60B was unable to interact with the native thin filaments unless CeTM and other associated proteins were removed by high-salt extraction. Purified CeTM was sufficient to restore the resistance of the salt extracted filaments from UNC-60B. In muscle cells, CeTM and UNC-60B were localized in different patterns. Suppression of CeTM by RNA interference resulted in disorganized actin filaments and paralyzed worms in wild-type background. However, in an ADF/cofilin mutant background, suppression of CeTM did not worsen actin organization and worm motility. These results suggest that tropomyosin is a physiological inhibitor of ADF/cofilin-dependent actin dynamics. PMID- 11901172 TI - The role of ARK in stress-induced apoptosis in Drosophila cells. AB - The molecular mechanisms of apoptosis are highly conserved throughout evolution. The homologs of genes essential for apoptosis in Caenorhabditis elegans and Drosophila melanogaster have been shown to be important for apoptosis in mammalian systems. Although a homologue for CED-4/apoptotic protease-activating factor (Apaf)-1 has been described in Drosophila, its exact function and the role of the mitochondrial pathway in its activation remain unclear. Here, we used the technique of RNA interference to dissect apoptotic signaling pathways in Drosophila cells. Inhibition of the Drosophila CED-4/Apaf-1-related killer (ARK) homologue resulted in pronounced inhibition of stress-induced apoptosis, whereas loss of ARK did not protect the cells from Reaper- or Grim-induced cell death. Reduction of DIAP1 induced rapid apoptosis in these cells, whereas the inhibition of DIAP2 expression did not but resulted in increased sensitivity to stress induced apoptosis; apoptosis in both cases was prevented by inhibition of ARK expression. Cells in which cytochrome c expression was decreased underwent apoptosis induced by stress stimuli, Reaper or Grim. These results demonstrate the central role of ARK in stress-induced apoptosis, which appears to act independently of cytochrome c. Apoptosis induced by Reaper or Grim can proceed via a distinct pathway, independent of ARK. PMID- 11901173 TI - The role of cytochrome c in caspase activation in Drosophila melanogaster cells. AB - The release of cytochrome c from mitochondria is necessary for the formation of the Apaf-1 apoptosome and subsequent activation of caspase-9 in mammalian cells. However, the role of cytochrome c in caspase activation in Drosophila cells is not well understood. We demonstrate here that cytochrome c remains associated with mitochondria during apoptosis of Drosophila cells and that the initiator caspase DRONC and effector caspase DRICE are activated after various death stimuli without any significant release of cytochrome c in the cytosol. Ectopic expression of the proapoptotic Bcl-2 protein, DEBCL, also fails to show any cytochrome c release from mitochondria. A significant proportion of cellular DRONC and DRICE appears to localize near mitochondria, suggesting that an apoptosome may form in the vicinity of mitochondria in the absence of cytochrome c release. In vitro, DRONC was recruited to a >700-kD complex, similar to the mammalian apoptosome in cell extracts supplemented with cytochrome c and dATP. These results suggest that caspase activation in insects follows a more primitive mechanism that may be the precursor to the caspase activation pathways in mammals. PMID- 11901174 TI - Antimicrobial polypeptides in host defense of the respiratory tract. PMID- 11901175 TI - Cytokines in innate host defense in the lung. PMID- 11901177 TI - What sensitized cells just might be doing in glomerulonephritis. PMID- 11901176 TI - The pulmonary collectins, SP-A and SP-D, orchestrate innate immunity in the lung. PMID- 11901178 TI - Serial killer: angiotensin drives cardiac hypertrophy via TGF-beta1. PMID- 11901179 TI - A chicken-or-egg conundrum in apoptosis: which comes first? Ceramide or PKCdelta? PMID- 11901180 TI - Splenic immunity and atherosclerosis: a glimpse into a novel paradigm? PMID- 11901181 TI - Deficiency of UDP-galactose:N-acetylglucosamine beta-1,4-galactosyltransferase I causes the congenital disorder of glycosylation type IId. AB - Deficiency of the Golgi enzyme UDP-Gal:N-acetylglucosamine beta-1,4 galactosyltransferase I (beta4GalT I) (E.C.2.4.1.38) causes a new congenital disorder of glycosylation (CDG), designated type IId (CDG-IId), a severe neurologic disease characterized by a hydrocephalus, myopathy, and blood-clotting defects. Analysis of oligosaccharides from serum transferrin by HPLC, mass spectrometry, and lectin binding revealed the loss of sialic acid and galactose residues. In skin fibroblasts and leukocytes, galactosyltransferase activity was reduced to 5% that of controls. In fibroblasts, a truncated polypeptide was detected that was about 12 kDa smaller in size than wild-type beta4GalT I and that failed to localize to the Golgi apparatus. Sequencing of the beta4GalT I cDNA and gene revealed an insertion of a single nucleotide (1031-1032insC) leading to premature translation stop and loss of the C-terminal 50 amino acids of the enzyme. The patient was homozygous and his parents heterozygous for this mutation. Expression of a corresponding mutant cDNA in COS-7 cells led to the synthesis of a truncated, inactive polypeptide, which localized to the endoplasmic reticulum. PMID- 11901182 TI - Cardiomyocyte overexpression of iNOS in mice results in peroxynitrite generation, heart block, and sudden death. AB - Increased inducible nitric oxide synthase (iNOS) expression is a component of the immune response and has been demonstrated in cardiomyocytes in septic shock, myocarditis, transplant rejection, ischemia, and dilated cardiomyopathy. To explore whether the consequences of such expression are adaptive or pathogenic, we have generated a transgenic mouse model conditionally targeting the expression of a human iNOS cDNA to myocardium. Chronic cardiac-specific upregulation of iNOS in transgenic mice led to increased production of peroxynitrite. This was associated with a mild inflammatory cell infiltrate, cardiac fibrosis, hypertrophy, and dilatation. While iNOS-overexpressing mice infrequently developed overt heart failure, they displayed a high incidence of sudden cardiac death due to bradyarrhythmia. This dramatic cardiac phenotype was rescued by specific attenuation of transgene activity. These data implicate cardiomyocyte iNOS overexpression as sufficient to cause cardiomyopathy, bradyarrhythmia, and sudden cardiac death. PMID- 11901183 TI - Protective immunity against atherosclerosis carried by B cells of hypercholesterolemic mice. AB - Atherosclerosis is characterized by vascular inflammation and associated with systemic and local immune responses to oxidized LDL (oxLDL) and other antigens. Since immunization with oxLDL reduces atherosclerosis, we hypothesized that the disease might be associated with development of protective immunity. Here we show that spleen-associated immune activity protects against atherosclerosis. Splenectomy dramatically aggravated atherosclerosis in hypercholesterolemic apoE knockout (apoE degrees ) mice. Transfer of spleen cells from atherosclerotic apoE degrees mice significantly reduced disease development in young apoE degrees mice. To identify the protective subset, donor spleen cells were divided into B and T cells by immunomagnetic separation before transfer. Protection was conferred by B cells, which reduced disease in splenectomized apoE degrees mice to one-fourth of that in splenectomized apoE degrees controls. Protection could also be demonstrated in intact, nonsplenectomized mice and was associated with an increase in antibody titers to oxLDL. Fewer CD4(+) T cells were found in lesions of protected mice, suggesting a role for T-B cell cooperation. These results demonstrate that B cell-associated protective immunity develops during atherosclerosis and reduces disease progression. PMID- 11901184 TI - Thymic dendritic cells traffic to thymi of allogeneic recipients and prolong graft survival. AB - We have demonstrated that murine thymic dendritic cells (DCs) isolated from donor mice have the capability to home to thymi of fully allogeneic recipients after intravenous injections, where they induce T cell deletions and prolong donor strain airway and skin graft survival. In contrast, infused splenic DCs immigrated poorly to thymi, and did not affect graft survival. These findings suggest that preferential homing may be an important mechanistic difference among subpopulations of DCs that mediate immune functions and illustrate a novel methodology that could have utility for induction of specific immunologic nonreactivity to allografts, or other disease-associated antigens. PMID- 11901185 TI - The alpha(1D)-adrenergic receptor directly regulates arterial blood pressure via vasoconstriction. AB - To investigate the physiological role of the alpha(1D)-adrenergic receptor (alpha(1D)-AR) subtype, we created mice lacking the alpha(1D)-AR (alpha(1D)(-/-)) by gene targeting and characterized their cardiovascular function. In alpha(1D)-/ mice, the RT-PCR did not detect any transcript of the alpha(1D)-AR in any tissue examined, and there was no apparent upregulation of other alpha(1)-AR subtypes. Radioligand binding studies showed that alpha(1)-AR binding capacity in the aorta was lost, while that in the heart was unaltered in alpha(1D)-/- mice. Non anesthetized alpha(1D)-/- mice maintained significantly lower basal systolic and mean arterial blood pressure conditions, relative to wild-type mice, and they showed no significant change in heart rate or in cardiac function, as assessed by echocardiogram. Besides hypotension, the pressor responses to phenylephrine and norepinephrine were decreased by 30-40% in alpha(1D)-/- mice. Furthermore, the contractile response of the aorta and the pressor response of isolated perfused mesenteric arterial beds to alpha(1)-AR stimulation were markedly reduced in alpha(1D)-/- mice. We conclude that the alpha(1D)-AR participates directly in sympathetic regulation of systemic blood pressure by vasoconstriction. PMID- 11901186 TI - Microtumor growth initiates angiogenic sprouting with simultaneous expression of VEGF, VEGF receptor-2, and angiopoietin-2. AB - Tumors have been thought to initiate as avascular aggregates of malignant cells that only later induce vascularization. Recently, this classic concept of tumor angiogenesis has been challenged by the suggestion that tumor cells grow by co opting preexisting host vessels and thus initiate as well-vascularized tumors without triggering angiogenesis. To discriminate between these two mechanisms, we have used intravital epifluorescence microscopy and multi-photon laser scanning confocal microscopy to visualize C6 microglioma vascularization and tumor cell behavior. To address the mechanisms underlying tumor initiation, we assessed the expression of VEGF, VEGF receptor-2 (VEGFR-2), and angiopoietin-2 (Ang-2), as well as endothelial cell proliferation. We show that multicellular aggregates (<< 1 mm(3)) initiate vascular growth by angiogenic sprouting via the simultaneous expression of VEGFR-2 and Ang-2 by host and tumor endothelium. Host blood vessels are not co-opted by tumor cells but rather are used as trails for tumor cell invasion of the host tissue. Our data further suggest that the established microvasculature of growing tumors is characterized by a continuous vascular remodeling, putatively mediated by the expression of VEGF and Ang-2. The results of this study suggest a new concept of vascular tumor initiation that may have important implications for the clinical application of antiangiogenic strategies. PMID- 11901187 TI - TGF-beta1 mediates the hypertrophic cardiomyocyte growth induced by angiotensin II. AB - Angiotensin II (Ang II), a potent hypertrophic stimulus, causes significant increases in TGFb1 gene expression. However, it is not known whether there is a causal relationship between increased levels of TGF-beta1 and cardiac hypertrophy. Echocardiographic analysis revealed that TGF-beta1-deficient mice subjected to chronic subpressor doses of Ang II had no significant change in left ventricular (LV) mass and percent fractional shortening during Ang II treatment. In contrast, Ang II-treated wild-type mice showed a >20% increase in LV mass and impaired cardiac function. Cardiomyocyte cross-sectional area was also markedly increased in Ang II-treated wild-type mice but unchanged in Ang II-treated TGF beta1-deficient mice. No significant levels of fibrosis, mitotic growth, or cytokine infiltration were detected in Ang II-treated mice. Atrial natriuretic factor expression was approximately 6-fold elevated in Ang II-treated wild-type, but not TGF-beta1-deficient mice. However, the alpha- to beta-myosin heavy chain switch did not occur in Ang II-treated mice, indicating that isoform switching is not obligatorily coupled with hypertrophy or TGF-beta1. The Ang II effect on hypertrophy was shown not to result from stimulation of the endogenous renin angiotensis system. These results indicate that TGF-beta1 is an important mediator of the hypertrophic growth response of the heart to Ang II. PMID- 11901188 TI - Bacterial induction of autoantibodies to beta2-glycoprotein-I accounts for the infectious etiology of antiphospholipid syndrome. AB - The antiphospholipid syndrome (APS) is characterized by the presence of pathogenic autoantibodies against beta2-glycoprotein-I (beta2GPI). The factors causing production of anti-beta2GPI remain unidentified, but an association with infectious agents has been reported. Recently, we identified a hexapeptide (TLRVYK) that is recognized specifically by a pathogenic anti-beta2GPI mAb. In the present study we evaluated the APS-related pathogenic potential of microbial pathogens carrying sequences related to this hexapeptide. Mice immunized with a panel of microbial preparations were studied for the development of anti-beta2GPI autoantibodies. IgG specific to the TLRVYK peptide were affinity purified from the immunized mice and passively infused intravenously into naive mice at day 0 of pregnancy. APS parameters were evaluated in the infused mice on day 15 of pregnancy. Following immunization, high titers of antipeptide [TLRVYK] anti beta2GPI Ab's were observed in mice immunized with Haemophilus influenzae, Neisseria gonorrhoeae, or tetanus toxoid. The specificity of binding to the corresponding target molecules was confirmed by competition and immunoblot assays. Naive mice infused with the affinity-purified antipeptide Ab's had significant thrombocytopenia, prolonged activated partial thromboplastin time and elevated percentage of fetal loss, similar to a control group of mice immunized with a pathogenic anti-beta2GPI mAb. Our study establishes a mechanism of molecular mimicry in experimental APS, demonstrating that bacterial peptides homologous with beta2GPI induce pathogenic anti-beta2GPI Ab's along with APS manifestations. PMID- 11901190 TI - Oxidation of the zinc-thiolate complex and uncoupling of endothelial nitric oxide synthase by peroxynitrite. AB - Nitric oxide (NO) is produced by NO synthase (NOS) in many cells and plays important roles in the neuronal, muscular, cardiovascular, and immune systems. In various disease conditions, all three types of NOS (neuronal, inducible, and endothelial) are reported to generate oxidants through unknown mechanisms. We present here the first evidence that peroxynitrite (ONOO(-)) releases zinc from the zinc-thiolate cluster of endothelial NOS (eNOS) and presumably forms disulfide bonds between the monomers. As a result, disruption of the otherwise SDS-resistant eNOS dimers occurs under reducing conditions. eNOS catalytic activity is exquisitely sensitive to ONOO(-), which decreases NO synthesis and increases superoxide anion (O(2)(.-)) production by the enzyme. The reducing cofactor tetrahydrobiopterin is not oxidized, nor does it prevent oxidation of eNOS by the same low concentrations of OONO(-). Furthermore, eNOS derived from endothelial cells exposed to elevated glucose produces more O(2)(.-), and, like eNOS purified from diabetic LDL receptor-deficient mice, contains less zinc and fewer SDS-resistant dimers. Hence, eNOS exposure to oxidants including ONOO(-) causes increased enzymatic uncoupling and generation of O(2)(.-) in diabetes, contributing further to endothelial cell oxidant stress. Regulation of the zinc thiolate center of NOS by ONOO(-) provides a novel mechanism for modulation of the enzyme function in disease. PMID- 11901189 TI - Acute intensive insulin therapy exacerbates diabetic blood-retinal barrier breakdown via hypoxia-inducible factor-1alpha and VEGF. AB - Acute intensive insulin therapy is an independent risk factor for diabetic retinopathy. Here we demonstrate that acute intensive insulin therapy markedly increases VEGF mRNA and protein levels in the retinae of diabetic rats. Retinal nuclear extracts from insulin-treated rats contain higher hypoxia-inducible factor-1alpha (HIF-1alpha) levels and demonstrate increased HIF-1alpha-dependent binding to hypoxia-responsive elements in the VEGF promoter. Blood-retinal barrier breakdown is markedly increased with acute intensive insulin therapy but can be reversed by treating animals with a fusion protein containing a soluble form of the VEGF receptor Flt; a control fusion protein has no such protective effect. The insulin-induced retinal HIF-1alpha and VEGF increases and the related blood-retinal barrier breakdown are suppressed by inhibitors of p38 mitogen activated protein kinase (MAPK) and phosphatidylinositol (PI) 3-kinase, but not inhibitors of p42/p44 MAPK or protein kinase C. Taken together, these findings indicate that acute intensive insulin therapy produces a transient worsening of diabetic blood-retinal barrier breakdown via an HIF-1alpha-mediated increase in retinal VEGF expression. Insulin-induced VEGF expression requires p38 MAPK and PI 3-kinase, whereas hyperglycemia-induced VEGF expression is HIF-1alpha-independent and requires PKC and p42/p44 MAPK. To our knowledge, these data are the first to identify a specific mechanism for the transient worsening of diabetic retinopathy, specifically blood-retinal barrier breakdown, that follows the institution of intensive insulin therapy. PMID- 11901192 TI - Expansion of pre-existing, lymph node-localized CD8+ T cells during supervised treatment interruptions in chronic HIV-1 infection. AB - To date, most studies have focused on the characterization of HIV-1-specific cellular immune responses in the peripheral blood (PB) of infected individuals. Much less is known about the comparative magnitude and breadth of responses in the lymphoid tissue. This study analyzed HIV-1-specific CD8+ T cell responses simultaneously in PB and lymph nodes (LNs) of persons with chronic HIV-1 infection and assessed the dynamics of these responses during antiretroviral treatment and supervised treatment interruption (STI). In untreated chronic infection, the magnitude of epitope-specific CD8+ T cell activity was significantly higher in LNs than in PB. Responses decreased in both compartments during highly active antiretroviral therapy, but this decline was more pronounced in PB. During STI, HIV-1-specific CD8+ T cell responses in PB increased significantly. Enhancement in breadth and magnitude was largely due to the expansion of pre-existing responses in the LNs, with new epitopes infrequently targeted. Taken together, these data demonstrate that HIV-1-specific CD8+ T cells are preferentially located in the LNs, with a subset of responses exclusively detectable in this compartment. Furthermore, the enhanced CD8+ T cell responses observed during STI in chronically infected individuals is largely due to expansion of pre-existing virus-specific CD8+ T cells, rather than the induction of novel responses. PMID- 11901191 TI - Protein kinase Cdelta amplifies ceramide formation via mitochondrial signaling in prostate cancer cells. AB - We studied the role of protein kinase C isoform PKCdelta in ceramide (Cer) formation, as well as in the mitochondrial apoptosis pathway induced by anticancer drugs in prostate cancer (PC) cells. Etoposide and paclitaxel induced Cer formation and apoptosis in PKCdelta-positive LNCaP and DU145 cells but not in PKCdelta-negative LN-TPA or PC-3 cells. In contrast, these drugs induced mitotic cell cycle arrest in all PC cell lines. Treatment with Rottlerin, a specific PKCdelta inhibitor, significantly inhibited drug-induced Cer formation and apoptosis in LNCaP cells, as did overexpression of dominant negative-type PKCdelta. Overexpression of wild-type PKCdelta had an opposite effect in PC-3 cells. Notably, etoposide induced biphasic Cer formation in LNCaP cells. The early and transient Cer increase resulted from de novo Cer synthesis, while the late and sustained Cer accumulation was derived from sphingomyelin hydrolysis by neutral sphingomyelinase (nSMase). Cer, in turn, induced mitochondrial translocation of PKCdelta and stimulated the activity of this kinase, promoting cytochrome c release and caspase-9 activation. Furthermore, the specific caspase 9 inhibitor LEHD-fmk significantly inhibited etoposide-induced nSMase activation, Cer accumulation, and PKCdelta mitochondrial translocation. These results indicate that PKCdelta plays a crucial role in activating anticancer drug-induced apoptosis signaling by amplifying the Cer-mediated mitochondrial amplification loop. PMID- 11901194 TI - Transient receptor potential 1 regulates capacitative Ca(2+) entry and Ca(2+) release from endoplasmic reticulum in B lymphocytes. AB - Capacitative Ca(2+) entry (CCE) activated by release/depletion of Ca(2+) from internal stores represents a major Ca(2+) influx mechanism in lymphocytes and other nonexcitable cells. Despite the importance of CCE in antigen-mediated lymphocyte activation, molecular components constituting this mechanism remain elusive. Here we demonstrate that genetic disruption of transient receptor potential (TRP)1 significantly attenuates both Ca(2+) release-activated Ca(2+) currents and inositol 1,4,5-trisphosphate (IP(3))-mediated Ca(2+) release from endoplasmic reticulum (ER) in DT40 B cells. As a consequence, B cell antigen receptor-mediated Ca(2+) oscillations and NF-AT activation are reduced in TRP1 deficient cells. Thus, our results suggest that CCE channels, whose formation involves TRP1 as an important component, modulate IP(3) receptor function, thereby enhancing functional coupling between the ER and plasma membrane in transduction of intracellular Ca(2+) signaling in B lymphocytes. PMID- 11901193 TI - Complement activation selectively potentiates the pathogenicity of the IgG2b and IgG3 isotypes of a high affinity anti-erythrocyte autoantibody. AB - By generating four IgG isotype-switch variants of the high affinity 34-3C anti erythrocyte autoantibody, and comparing them to the IgG variants of the low affinity 4C8 anti-erythrocyte autoantibody that we have previously studied, we evaluated in this study how high affinity binding to erythrocytes influences the pathogenicity of each IgG isotype in relation to the respective contributions of Fcgamma receptor (FcgammaR) and complement. The 34-3C autoantibody opsonizing extensively circulating erythrocytes efficiently activated complement in vivo (IgG2a = IgG2b > IgG3), except for the IgG1 isotype, while the 4C8 IgG autoantibody failed to activate complement. The pathogenicity of the 34-3C autoantibody of IgG2b and IgG3 isotypes was dramatically higher (>200-fold) than that of the corresponding isotypes of the 4C8 antibody. This enhanced activity was highly (IgG2b) or totally (IgG3) dependent on complement. In contrast, erythrocyte-binding affinities only played a minor role in in vivo hemolytic activities of the IgG1 and IgG2a isotypes of 34-3C and 4C8 antibodies, where complement was not or only partially involved, respectively. The remarkably different capacities of four different IgG isotypes of low and high affinity anti erythrocyte autoantibodies to activate FcgammaR-bearing effector cells and complement in vivo demonstrate the role of autoantibody affinity maturation and of IgG isotype switching in autoantibody-mediated pathology. PMID- 11901195 TI - Major histocompatibility complex class II presentation of cell-associated antigen is mediated by CD8alpha+ dendritic cells in vivo. AB - Antigen-specific B cells express major histocompatibility complex class II and can present antigen directly to T cells. Adoptive transfer experiments using transgenic B and T cells demonstrated that antigen-specific B cells can also efficiently transfer antigen to another cell for presentation to T cells in vivo. To identify the antigen-presenting cell that receives antigens from B cells, a strategy was developed to follow the traffic of B cell-derived proteins in vivo. B cells were labeled with the fluorescent dye CFSE and loaded with antigen, before adoptive transfer into recipient mice. Populations of splenocytes from the recipient mice were later assayed for the presence of fluorescent proteins and for the ability to activate T cells. A small number of CD8alpha+CD4-CD11b(lo) dendritic cells (DCs) contain proteins transferred from B cells and these DCs effectively present antigens derived from the B cells to T cells. The results suggest that CD8alpha+ DCs sample the cells and membranes in their environment for presentation to T cells circulating through the T cell zone. This function of CD8alpha+ DCs may be relevant to the priming of an immune response or the induction of T cell tolerance. PMID- 11901196 TI - Generation of human CD8 T regulatory cells by CD40 ligand-activated plasmacytoid dendritic cells. AB - Although CD8 T cell-mediated immunosuppression has been a well-known phenomenon during the last three decades, the nature of primary CD8 T suppressor cells and the mechanism underlying their generation remain enigmatic. We demonstrated that naive CD8 T cells primed with allogeneic CD40 ligand-activated plasmacytoid dendritic cells (DC)2 differentiated into CD8 T cells that displayed poor secondary proliferative and cytolytic responses. By contrast, naive CD8 T cells primed with allogeneic CD40 ligand-activated monocyte-derived DCs (DC1) differentiated into CD8 T cells, which proliferated to secondary stimulation and killed allogeneic target cells. Unlike DC1-primed CD8 T cells that produced large amounts of interferon (IFN)-gamma upon restimulation, DC2-primed CD8 T cells produced significant amounts of interleukin (IL)-10, low IFN-gamma, and no IL-4, IL-5, nor transforming growth factor (TGF)-beta. The addition of anti-IL-10 neutralizing monoclonal antibodies during DC2 and CD8 T cell coculture, completely blocked the generation of IL-10-producing anergic CD8 T cells. IL-10 producing CD8 T cells strongly inhibit the allospecific proliferation of naive CD8 T cells to monocytes, and mature and immature DCs. This inhibition was mediated by IL-10, but not by TGF-beta. IL-10-producing CD8 T cells could inhibit the bystander proliferation of naive CD8 T cells, provided that they were restimulated nearby to produce IL-10. IL-10-producing CD8 T cells could not inhibit the proliferation of DC1-preactivated effector T cells. This study demonstrates that IL-10-producing CD8 T cells are regulatory T cells, which provides a cellular basis for the phenomenon of CD8 T cell-mediated immunosuppression and suggests a role for plasmacytoid DC2 in immunological tolerance. PMID- 11901197 TI - Differential requirement for LAT and SLP-76 in GPVI versus T cell receptor signaling. AB - Mice deficient in the adaptor Src homology 2 domain-containing leukocyte phosphoprotein of 76 kD (SLP-76) exhibit a bleeding disorder and lack T cells. Linker for activation of T cells (LAT)-deficient mice exhibit a similar T cell phenotype, but show no signs of hemorrhage. Both SLP-76 and LAT are important for optimal platelet activation downstream of the collagen receptor, GPVI. In addition, SLP-76 is involved in signaling mediated by integrin alphaIIbbeta3. Because SLP-76 and LAT function coordinately in T cell signal transduction, yet their roles appear to differ in hemostasis, we investigated in detail the functional consequences of SLP-76 and LAT deficiencies in platelets. Previously we have shown that LAT(-/-) platelets exhibit defective responses to the GPVI specific agonist, collagen-related peptide (CRP). Consistent with this, we find that surface expression of P-selectin in response to high concentrations of GPVI ligands is reduced in both LAT- and SLP-76-deficient platelets. However, platelets from LAT(-/-) mice, but not SLP-76(-/-) mice, aggregate normally in response to high concentrations of collagen and convulxin. Additionally, unlike SLP-76, LAT is not tyrosine phosphorylated after fibrinogen binding to integrin alphaIIbbeta3, and collagen-stimulated platelets deficient in LAT spread normally on fibrinogen-coated surfaces. Together, these findings indicate that while LAT and SLP-76 are equally required for signaling via the T cell antigen receptor (TCR) and pre-TCR, platelet activation downstream of GPVI and alphaIIbbeta3 shows a much greater dependency on SLP-76 than LAT. PMID- 11901198 TI - Progress toward a human CD4/CCR5 transgenic rat model for de novo infection by human immunodeficiency virus type 1. AB - The development of a permissive small animal model for the study of human immunodeficiency virus type (HIV)-1 pathogenesis and the testing of antiviral strategies has been hampered by the inability of HIV-1 to infect primary rodent cells productively. In this study, we explored transgenic rats expressing the HIV 1 receptor complex as a susceptible host. Rats transgenic for human CD4 (hCD4) and the human chemokine receptor CCR5 (hCCR5) were generated that express the transgenes in CD4(+) T lymphocytes, macrophages, and microglia. In ex vivo cultures, CD4(+) T lymphocytes, macrophages, and microglia from hCD4/hCCR5 transgenic rats were highly susceptible to infection by HIV-1 R5 viruses leading to expression of abundant levels of early HIV-1 gene products comparable to those found in human reference cultures. Primary rat macrophages and microglia, but not lymphocytes, from double-transgenic rats could be productively infected by various recombinant and primary R5 strains of HIV-1. Moreover, after systemic challenge with HIV-1, lymphatic organs from hCD4/hCCR5 transgenic rats contained episomal 2-long terminal repeat (LTR) circles, integrated provirus, and early viral gene products, demonstrating susceptibility to HIV-1 in vivo. Transgenic rats also displayed a low-level plasma viremia early in infection. Thus, transgenic rats expressing the appropriate human receptor complex are promising candidates for a small animal model of HIV-1 infection. PMID- 11901199 TI - Short-lived and long-lived bone marrow plasma cells are derived from a novel precursor population. AB - The contribution that long-lived bone marrow (BM) plasma cells (PCs) provide to enduring humoral immunity has been underscored by a number of recent studies. However, little is known about the immediate precursors that give rise to long lived PCs in the BM of immune individuals. We have identified subsets of antigen experienced B cells within the immune BM that are precursors to PCs. These PC precursors arise in the BM 14 days after immunization and persist for greater than 9 months. Phenotypically distinct subsets of PC precursors give rise to short-lived or long-lived PCs. The differentiation of PC precursors to PCs occurs in the absence of antigen and requires cell division. The functional significance of these newly identified PC precursors in the persistence and quality of the humoral immune response is discussed. PMID- 11901200 TI - Epidermal transglutaminase (TGase 3) is the autoantigen of dermatitis herpetiformis. AB - Gluten sensitivity typically presents as celiac disease, a common chronic small intestinal disorder. However, in certain individuals it is associated with dermatitis herpetiformis, a blistering skin disease characterized by granular IgA deposits in the papillary dermis. While tissue transglutaminase has been implicated as the major autoantigen of gluten sensitive disease, there has been no explanation as to why this condition appears in two distinct forms. Here we show that while sera from patients with either form of gluten sensitive disease react both with tissue transglutaminase and the related enzyme epidermal (type 3) transglutaminase, antibodies in patients having dermatitis herpetiformis show a markedly higher avidity for epidermal transglutaminase. Further, these patients have an antibody population specific for this enzyme. We also show that the IgA precipitates in the papillary dermis of patients with dermatitis herpetiformis, the defining signs of the disease, contain epidermal transglutaminase, but not tissue transglutaminase or keratinocyte transglutaminase. These findings demonstrate that epidermal transglutaminase, rather than tissue transglutaminase, is the dominant autoantigen in dermatitis herpetiformis and explain why skin symptoms appear in a proportion of patients having gluten sensitive disease. PMID- 11901201 TI - Polycomb group gene rae28 is required for sustaining activity of hematopoietic stem cells. AB - The rae28 gene (rae28), also designated as mph1, is a mammalian ortholog of the Drosophila polyhomeotic gene, a member of Polycomb group genes (PcG). rae28 constitutes PcG complex 1 for maintaining transcriptional states which have been once initiated, presumably through modulation of the chromatin structure. Hematopoietic activity was impaired in the fetal liver of rae28-deficient animals (rae28-/-), as demonstrated by progressive reduction of hematopoietic progenitors of multilineages and poor expansion of colony forming units in spleen (CFU-S(12)) during embryonic development. An in vitro long-term culture-initiating cell assay suggested a reduction in hematopoietic stem cells (HSCs), which was confirmed in vivo by reconstitution experiments in lethally irradiated congenic recipient mice. The competitive repopulating units (CRUs) reflect HSCs supporting multilineage blood-cell production. CRUs were generated, whereas the number of CRUs was reduced by a factor of 20 in the rae28-/- fetal liver. We also performed serial transplantation experiments to semiquantitatively measure self-renewal activity of CRUs in vivo. Self-renewal activity of CRUs was 15-fold decreased in rae28-/-. Thus the compromised HSCs were presumed to reduce hematopoietic activity in the rae28-/- fetal liver. This is the first report to suggest that rae28 has a crucial role in sustaining the activity of HSCs to maintain hematopoiesis. PMID- 11901202 TI - B-1a B cells that link the innate and adaptive immune responses are lacking in the absence of the spleen. AB - Splenectomized individuals are prone to overwhelming infections with encapsulated bacteria and splenectomy of mice increases susceptibility to streptococcal infections, yet the exact mechanism by which the spleen protects against such infections is unknown. Using congenitally asplenic mice as a model, we show that the spleen is essential for the generation of B-1a cells, a B cell population that cooperates with the innate immune system to control early bacterial and viral growth. Splenectomy of wild-type mice further demonstrated that the spleen is also important for the survival of B-1a cells. Transfer experiments demonstrate that lack of these cells, as opposed to the absence of the spleen per se, is associated with an inability to mount a rapid immune response against streptococcal polysaccharides. Thus, absence of the spleen and the associated increased susceptibility to streptococcal infections is correlated with lack of B 1a B cells. These findings reveal a hitherto unknown role of the spleen in generating and maintaining the B-1a B cell pool. PMID- 11901203 TI - Pharmacological stimulation of the cholinergic antiinflammatory pathway. AB - Efferent activity in the vagus nerve can prevent endotoxin-induced shock by attenuating tumor necrosis factor (TNF) synthesis. Termed the "cholinergic antiinflammatory pathway," inhibition of TNF synthesis is dependent on nicotinic alpha-bungarotoxin-sensitive acetylcholine receptors on macrophages. Vagus nerve firing is also stimulated by CNI-1493, a tetravalent guanylhydrazone molecule that inhibits systemic inflammation. Here, we studied the effects of pharmacological and electrical stimulation of the intact vagus nerve in adult male Lewis rats subjected to endotoxin-induced shock to determine whether intact vagus nerve signaling is required for the antiinflammatory action of CNI-1493. CNI-1493 administered via the intracerebroventricular route was 100,000-fold more effective in suppressing endotoxin-induced TNF release and shock as compared with intravenous dosing. Surgical or chemical vagotomy rendered animals sensitive to TNF release and shock, despite treatment with CNI-1493, indicating that an intact cholinergic antiinflammatory pathway is required for antiinflammatory efficacy in vivo. Electrical stimulation of either the right or left intact vagus nerve conferred significant protection against endotoxin-induced shock, and specifically attenuated serum and myocardial TNF, but not pulmonary TNF synthesis, as compared with sham-operated animals. Together, these results indicate that stimulation of the cholinergic antiinflammatory pathway by either pharmacological or electrical methods can attenuate the systemic inflammatory response to endotoxin-induced shock. PMID- 11901204 TI - Two subsets of naive T helper cells with distinct T cell receptor excision circle content in human adult peripheral blood. AB - During ageing thymic function declines and is unable to meet the demand for peripheral T helper (Th) cell replenishment. Therefore, population maintenance of naive Th cells must be at least partly peripherally based. Such peripheral postthymic expansion of recent thymic emigrants (RTEs) during ageing consequently should lead to loss or dilution of T cell receptor excision circles (TRECs) from a subset of naive T cells. We have identified two subsets of naive Th cells in human adult peripheral blood characterized by a striking unequal content of TRECs, indicating different peripheral proliferative histories. TRECs are highly enriched in peripheral naive CD45RA(+) Th cells coexpressing CD31 compared with peripheral naive CD45RA(+) Th cells lacking CD31 expression, in which TRECs can hardly be detected. Furthermore we show that CD31(-)CD45RA(+) Th cells account for increasing percentages of the naive peripheral Th cell pool during ageing but retain phenotypic and functional features of naive Th cells. As CD31 is lost upon T cell receptor (TCR) engagement in vitro, we hypothesize that TCR triggering is a prerequisite for homeostatically driven peripheral postthymic expansion of human naive RTEs. We describe here the identification of peripherally expanded naive Th cells in human adult blood characterized by the loss of CD31 expression and a highly reduced TREC content. PMID- 11901207 TI - Deactivation of vasodilator responses by GRK2 overexpression: a mechanism or the mechanism for hypertension? PMID- 11901206 TI - Harnessing a neural-immune circuit to control inflammation and shock. PMID- 11901205 TI - Modulation of LIGHT-HVEM costimulation prolongs cardiac allograft survival. AB - LIGHT (TNFSF14), a tumor necrosis factor superfamily member expressed by activated T cells, binds to herpes virus entry mediator (HVEM) which is constitutively expressed by T cells and costimulates T cell activation in a CD28 independent manner. Given interest in regulating the effector functions of T cells in vivo, we examined the role of LIGHT-HVEM costimulation in a murine cardiac allograft rejection model. Normal hearts lacked LIGHT or HVEM mRNA expression, but allografts showed strong expression of both genes from day 3 after transplant, and in situ hybridization and immunohistology-localized LIGHT and HVEM to infiltrating leukocytes. To test the importance of LIGHT expression on allograft survival, we generated LIGHT-/- mice by homologous recombination. The mean survival of fully major histocompatibility complex-mismatched vascularized cardiac allografts in LIGHT-/- mice (10 days, P < 0.05) or cyclosporine A (CsA)-treated LIGHT+/+ mice (10 days, P < 0.05) was only slightly prolonged compared with LIGHT+/+ mice (7 days). However, mean allograft survival in CsA-treated LIGHT-/- allograft recipients (30 days) was considerably enhanced (P < 0.001) compared with the 10 days of mean survival in either untreated LIGHT /- mice or CsA-treated LIGHT+/+ controls. Molecular analyzes showed that the beneficial effects of targeting of LIGHT in CsA-treated recipients were accompanied by decreased intragraft expression of interferon (IFN)-gamma, plus IFN-gamma-induced chemokine, inducible protein-10, and its receptor, CXCR3. Treatment of LIGHT+/+ allograft recipients with HVEM-Ig plus CsA also enhanced mean allograft survival (21 days) versus wild-type controls receiving HVEM-Ig (mean of 7 days) or CsA alone (P < 0.001). Our data suggest that T cell to T cell mediated LIGHT/HVEM-dependent costimulation is a significant component of the host response leading to cardiac allograft rejection. PMID- 11901208 TI - p75-nerve growth factor as an antiapoptotic complex: independence versus cooperativity in protection from enediyne chemotherapeutic agents. AB - Growth factors, including nerve growth factor (NGF), have been hypothesized to play a role in resistance to chemotherapeutic agent-induced apoptosis. Induction by NGF of resistance to apoptosis is primarily thought to be the result of its binding to its high-affinity receptor, TrkA. The low-affinity NGF receptor, p75, has long been thought merely to facilitate NGF binding to TrkA. However, we have previously shown that the binding of NGF to its low-affinity receptor, p75, protects neuroblastoma cells that do not express TrkA against apoptosis induced by enediyne chemotherapeutic agents. In cells that express both receptors, it is not clear what determines which receptor is responsible for the protective effect of NGF. We now show that, in enediyne-treated SH-SY5Y neuroblastoma transfectants with native levels of p75 and a low TrkA/p75 ratio (1/100), the anti-apoptotic effect of NGF requires binding to p75. In contrast, in transfectants with native levels of p75 and a high TrkA/p75 ratio (100/100), NGF treatment prevents enediyne-induced apoptosis by a mechanism independent of p75 binding. Treatment of low TrkA/p75 ratio cells with NGF results in activation and nuclear translocation of NF-kappaB and tyrosine phosphorylation of TrkA. Analogous treatment of high TrkA/p75 ratio cells results only in phosphorylation of TrkA even though nuclear factor (NF)-kappaB signaling is not inactive and can be initiated by other ligands. The ratio of TrkA/p75 in cells that express both receptors probably contributes to the determination of which of the two known roles of p75 (i.e., TrkA independent or TrkA facilitatory) are responsible for NGF-mediated protection from enediyne-induced apoptosis. PMID- 11901209 TI - Identification of a potent and selective pharmacophore for Cdc25 dual specificity phosphatase inhibitors. AB - Small molecules provide powerful tools to interrogate biological pathways but many important pathway participants remain refractory to inhibitors. For example, Cdc25 dual-specificity phosphatases regulate mammalian cell cycle progression and are implicated in oncogenesis, but potent and selective inhibitors are lacking for this enzyme class. Thus, we evaluated 10,070 compounds in a publicly available chemical repository of the National Cancer Institute for in vitro inhibitory activity against oncogenic, full-length, recombinant human Cdc25B. Twenty-one compounds had mean inhibitory concentrations of <1 microM; >75% were quinones and >40% were of the para-naphthoquinone structural type. Most notable was NSC 95397 (2,3-bis-[2-hydroxyethylsulfanyl]-[1,4]naphthoquinone), which displayed mixed inhibition kinetics with in vitro K(i) values for Cdc25A, -B, and -C of 32, 96, and 40 nM, respectively. NSC 95397 was more potent than any inhibitor of dual specificity phosphatases described previously and 125- to 180 fold more selective for Cdc25A than VH1-related dual-specificity phosphatase or protein tyrosine phosphatase 1b, respectively. Modification of the bis thioethanol moiety markedly decreased enzyme inhibitory activity, indicating its importance for bioactivity. NSC 95397 showed significant growth inhibition against human and murine carcinoma cells and blocked G(2)/M phase transition. A potential Cdc25 site of interaction was postulated based on molecular modeling with these quinones. We propose that inhibitors based on this chemical structure could serve as useful tools to probe the biological function of Cdc25. PMID- 11901210 TI - Transport of amino acid-related compounds mediated by L-type amino acid transporter 1 (LAT1): insights into the mechanisms of substrate recognition. AB - The L-type amino acid transporter 1 (LAT1) is an Na(+)-independent neutral amino acid transporter subserving the amino acid transport system L. Because of its broad substrate selectivity, system L has been proposed to be responsible for the permeation of amino acid-related drugs through the plasma membrane. To understand the mechanisms of substrate recognition, we have examined the LAT1-mediated transport using a Xenopus laevis oocyte expression system. LAT1-mediated [(14)C]phenylalanine uptake was strongly inhibited in a competitive manner by aromatic-amino acid derivatives including L-dopa, alpha-methyldopa, melphalan, triiodothyronine, and thyroxine, whereas phenylalanine methyl ester, N-methyl phenylalanine, dopamine, tyramine, carbidopa, and droxidopa did not inhibit [(14)C]phenylalanine uptake. Gabapentin, a gamma-amino acid, also exerted a competitive inhibition on LAT1-mediated [(14)C]phenylalanine uptake. Although most of the compounds that inhibited LAT1-mediated uptake were able to induce the efflux of [(14)C]phenylalanine preloaded to the oocytes expressing LAT1 through the obligatory exchange mechanism, melphalan, triiodothyronine, and thyroxine did not induce the significant efflux. Based on the experimental and semiempirical computational analyses, it is proposed that, for an aromatic amino acid to be a LAT1 substrate, it must have a free carboxyl and an amino group. The carbonyl oxygen closer to the amino group needs a computed charge of -0.55 approximately 0.56 and must not participate in hydrogen bonding. In addition, the hydrophobic interaction between the substrate side chain and the substrate binding site of LAT1 seems to be crucial for the substrate binding. A substrate, however, becomes a blocker once Connolly accessible areas become large and/or the molecule has a high calculated logP value, such as those for melphalan, triiodothyronine, and thyroxine. PMID- 11901211 TI - Overexpression of the ubiquitin-conjugating enzyme Cdc34 confers resistance to methylmercury in Saccharomyces cerevisiae. AB - A search was made for genes that confer resistance to methylmercury in yeast using a genomic DNA library derived from Saccharomyces cerevisiae. The genomic library was introduced into yeast and transformants that grew in the presence of a normally toxic concentration of methylmercury were selected. We sequenced the genomic DNA fragment in the plasmid from the clone with the highest resistance to methylmercury and analyzed the sequence for presence of an open reading frame that might confer resistance to methylmercury. We identified a gene, CDC34 (also known as UBC3), that increased resistance to methylmercury when overexpressed in yeast. CDC34 encodes a ubiquitin-conjugating enzyme; such proteins play important roles in the selective targeting of proteins for degradation. Overexpression of UBC4 and of UBC7, two other genes for ubiquitin-conjugating enzymes, also conferred resistance to methylmercury. Yeast strains transformed with the CDC34 gene were resistant not only to methylmercury but also to mercuric chloride and p chloromercuribenzoate. To our knowledge, this is the first demonstration that overexpression of genes for ubiquitin-conjugating enzymes confers resistance to xenobiotics. Our results suggest that ubiquitination system might be involved in protection against the toxicity of mercury compounds, such as methylmercury, in eukaryotic cells. PMID- 11901212 TI - Induction of biphasic DNA double strand breaks and activation of multiple repair protein complexes by DNA topoisomerase I drug 7-ethyl-10-hydroxy-camptothecin. AB - Camptothecins demonstrate a broad spectrum of antitumor activity. Although they are known to trap DNA topoisomerase I on DNA, form cleavable complexes, and generate DNA breaks upon collision with DNA or RNA polymerases, the precise mechanisms predictive for antitumor activity remain to be identified. Recent studies using panels of colorectal and breast cancer cell lines indicate that events downstream of cleavable complexes are more relevant. In this study, we chose SN-38, an active metabolite of irinotecan, to characterize DNA double strand breaks and repair mechanisms induced by this type of drugs using a human head and neck squamous cell carcinoma cell line A253. The results showed that 2-h exposure of cells to an IC(50) concentration of SN-38 induces biphasic DNA double strand break (DSBs): an immediate phase, which was greatly reduced within 8 h, and a lagging phase, culminating 24 h after drug removal. Three DNA double-strand break repair protein complexes were activated: DNA-dependent protein kinase (DNA PK), NBS1-MRE11-RAD50, and BRCA1. Aphidicolin, a DNA polymerase inhibitor, abolished both phase I DSBs and the activation of repair protein complexes, suggesting that they resulted from the collision between the cleavable complex and DNA polymerase of S-phase cells. This is in contrast to ionizing radiation induced activation of DNA-PK and NBS1-MRE11-RAD50 complexes that occur predominantly among non-S-phase cells. The trigger for phase II DSBs cannot be abolished by aphidicolin. The data also indicate that DNA fragments in the size of 50 to 200 kilobases were detected in the lagging phase. This suggests that the late DNA DSBs were associated with apoptotic cell death. PMID- 11901213 TI - Vascular-targeted overexpression of G protein-coupled receptor kinase-2 in transgenic mice attenuates beta-adrenergic receptor signaling and increases resting blood pressure. AB - Cardiovascular regulation is tightly controlled by signaling through G protein coupled receptors (GPCRs). beta-Adrenergic receptors (ARs) are GPCRs that regulate inotropy and chronotropy in the heart and mediate vasodilation, which critically influences systemic vascular resistance. GPCR kinases (GRKs), including GRK2 (or betaARK1), phosphorylate and desensitize agonist-activated betaARs. Myocardial GRK2 levels are increased in heart failure and data suggest that vascular levels may also be elevated in hypertension. Therefore, we generated transgenic mice with vascular smooth muscle (VSM) targeted overexpression of GRK2, using a portion of the SM22alpha promoter, to determine its impact on vascular betaAR regulation. VSM betaAR signaling, as determined by adenylyl cyclase and mitogen-activated protein (MAP) kinase activation assays, was attenuated when GRK2 was overexpressed 2- to 3-fold. In vivo vasodilation in response to betaAR stimulation using isoproterenol was attenuated and conscious resting mean arterial blood pressure was elevated from 96 +/- 2 mm Hg in nontransgenic littermate control (NLC) mice (n = 9) to 112 +/- 3 mm Hg and 117 +/ 2 mm Hg in two different lines of SM22alpha-GRK2 transgenic mice (n = 7 and n = 5, respectively; p < 0.05). Interestingly, medial VSM thickness was increased 30% from 29.8 +/- 1.6 microm in NLC mice (n = 6) to 39.4 +/- 1.6 microm in SM22alpha GRK2 mice (n = 7) (p < 0.05) and vascular GRK2 overexpression was sufficient to cause cardiac hypertrophy. These data indicate that we have developed a unique mouse model of hypertension, providing insight into the contribution that vascular betaAR signaling makes toward resting blood pressure and overall cardiovascular regulation. Moreover, they suggest that GRK2 plays an important role in vascular control and may represent a novel therapeutic target for hypertension. PMID- 11901214 TI - Novel "nonkinase" phorbol ester receptors: the C1 domain connection. AB - In recent years, there have been great advances in our understanding of the pharmacology and biology of the receptors for the phorbol ester tumor promoters and the second messenger diacylglycerol (DAG). The traditional view of protein kinase C (PKC) as the sole receptor for the phorbol esters has been challenged with the discovery of proteins unrelated to PKC that bind phorbol esters with high affinity, suggesting a high degree of complexity in the signaling pathways activated by DAG. These novel "nonkinase" phorbol ester receptors include chimaerins (a family of Rac GTPase activating proteins), RasGRPs (exchange factors for Ras/Rap1), and Munc13 isoforms (scaffolding proteins involved in exocytosis). In all cases, phorbol ester binding occurs at the single C1 domain present in these proteins and, as in PKC isozymes, ligand binding is a phospholipid-dependent event. Moreover, the novel phorbol ester receptors are also subject to subcellular redistribution or "translocation" by phorbol esters, leading to their association to different effector and/or regulatory molecules. Clearly, the use of phorbol esters as specific activators of PKC in cellular models is questionable. Alternative pharmacological and molecular approaches are therefore needed to dissect the involvement of each receptor class as a mediator of phorbol ester/DAG responses. PMID- 11901216 TI - Up-regulation of tryptophan hydroxylase expression and serotonin synthesis by sertraline. AB - The neurotransmitter serotonin is involved in a variety of brain functions, and abnormal changes in serotonin neurotransmission are associated with an array of psychiatric disorders, including depression. Sertraline is a selective serotonin reuptake inhibitor (SSRI) and an effective antidepressant. Sertraline increases the serotonin concentration in the synaptic cleft by a short-term action; however, clinical improvement is observed only after several weeks, suggesting that the therapeutic effect may be caused by long-term alterations in serotonin transmission. We determined the effects of sertraline on serotonin synthesis in vivo and in vitro. Long-term treatment of rats with sertraline up-regulated mRNA and protein levels of the serotonin-synthesizing enzyme tryptophan hydroxylase (TPH), as determined by in situ hybridization and immunocytochemistry, respectively. In vitro studies using RBL-2H3 cells also showed an increase in mRNA and protein levels of TPH by sertraline, as determined by Northern blot and immunoblot analyses, respectively. This was accompanied by increases in the levels of TPH enzymatic activity and total serotonin. These data demonstrate that in addition to the known short-term action as an uptake blocker, sertraline also exerts a long-term effect on the serotonin neurotransmission by enhancing serotonin synthesis. A similar effect was observed with another SSRI, fluoxetine, but not with the non-SSRI chlorpromazine. The up-regulation of TPH gene expression by sertraline was attenuated by the protein kinase A (PKA) inhibitor N [2-(p-bromocinnamylamine)-ethyl]-5-isoquinolinesulfonamine, suggesting that a mechanism involving the PKA signaling pathway might at least in part mediate the long-term therapeutic action. PMID- 11901215 TI - Side-chain substitutions within angiotensin II reveal different requirements for signaling, internalization, and phosphorylation of type 1A angiotensin receptors. AB - Binding of the peptide hormone angiotensin II (AngII) to the type 1 (AT(1A)) receptor and the subsequent activation of phospholipase C-mediated signaling, involves specific determinants within the AngII peptide sequence. In contrast, the contribution of such determinants to AT(1A) receptor internalization, phosphorylation and activation of mitogen-activated protein kinase (MAPK) signaling is not known. In this study, the internalization of an enhanced green fluorescent protein-tagged AT(1A) receptor (AT(1A)-EGFP), in response to AngII and a series of substituted analogs, was visualized and quantified using confocal microscopy. AngII-stimulation resulted in a rapid, concentration-dependent internalization of the chimeric receptor, which was prevented by pretreatment with the nonpeptide AT(1) receptor antagonist EXP3174. Remarkably, AT(1A) receptor internalization was unaffected by substitution of AngII side chains, including single and double substitutions of Tyr(4) and Phe(8) that abolish phospholipase C signaling through the receptor. AngII-induced receptor phosphorylation was significantly inhibited by several substitutions at Phe(8) as well as alanine replacement of Asp(1). The activation of MAPK was only significantly inhibited by substitutions at position eight in the peptide and specific substitutions did not equally inhibit inositol phosphate production, receptor phosphorylation and MAPK activation. These results indicate that separate, yet overlapping, contacts made between the AngII peptide and the AT(1A) receptor select/induce distinct receptor conformations that preferentially affect particular receptor outcomes. The requirements for AT(1A) receptor internalization seem to be less stringent than receptor activation and signaling, suggesting an inherent bias toward receptor deactivation. PMID- 11901217 TI - Arachidonic acid differentially affects basal and lipopolysaccharide-induced sPLA(2)-IIA expression in alveolar macrophages through NF-kappaB and PPAR-gamma dependent pathways. AB - Secretory type IIA phospholipase A(2) (sPLA(2)-IIA) is a critical enzyme involved in inflammatory diseases. We have previously identified alveolar macrophages (AMs) as the major pulmonary source of lipopolysaccharide (LPS)-induced sPLA(2) IIA expression in a guinea pig model of acute lung injury (ALI). Here, we examined the role of arachidonic acid (AA) in the regulation of basal and LPS induced sPLA(2)-IIA expression in AMs. We showed that both AA and its nonmetabolizable analog, 5,8,11,14-eicosatetraynoic acid (ETYA), inhibited sPLA(2)-IIA synthesis in unstimulated AMs. However, only AA inhibited sPLA(2)-IIA expression in LPS-stimulated cells, suggesting that this effect requires metabolic conversion of AA. Indeed, cyclooxygenase inhibitors abolished this down regulation. Prostaglandins PGE(2), PGA(2), and 15d-PGJ(2) also inhibited the LPS induced sPLA(2)-IIA expression. Nuclear factor-kappaB (NF-kappaB) was found to regulate sPLA(2)-IIA expression in AMs. Both AA and ETYA inhibited basal activation of NF-kappaB but had no effect on LPS-induced NF-kappaB translocation, suggesting that suppression of sPLA(2)-IIA synthesis by AA in LPS-stimulated cells occurs via a NF-kappaB-independent pathway. 15-Deoxy-Delta(12,14)-PGJ(2) and ciglitazone, which are, respectively, natural and synthetic ligands for peroxisome proliferator-activated receptor-gamma (PPAR-gamma), inhibited LPS induced sPLA(2)-IIA synthesis, whereas PPAR-alpha ligands were ineffective. Moreover, electrophoretic mobility shift assay showed PPAR activation by AA and PPAR-gamma ligands in LPS-stimulated AMs. Our results suggest that the down regulation of basal sPLA(2)-IIA expression is unrelated to the metabolic conversion of AA but is dependent on the impairment of NF-kappaB activation. In contrast, the inhibition of LPS-stimulated sPLA(2)-IIA expression is mediated by cyclooxygenase-derived metabolites of AA and involves a PPAR-gamma-dependent pathway. These findings provide new insights for the treatment of ALI. PMID- 11901218 TI - Heterogeneous nuclear ribonucleoprotein A1 and regulation of the xenobiotic inducible gene Cyp2a5. AB - The heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) functions in the packaging of nascent RNA polymerase II transcripts and participates in a variety of nuclear and cytoplasmic processes that modulate gene expression. The RNA binding characteristics of hnRNP A1 suggest that it can modulate the expression of specific genes, but little is known about its possible targets in vivo. In this article, we show that hnRNP A1 interacts with the transcript of a cytochrome P450 gene, Cyp2a5, induced by xenobiotics and during liver damage. Binding of the hnRNP A1 to CYP2A5 mRNA was demonstrated by immunoprecipitation of the xenobiotic stimulated (37/39 kDa) CYP2A5 mRNA-protein complex with a monoclonal anti-hnRNP A1 antibody, by partial trypsin digestion of the complex, and by showing that the RNA-protein complex is not formed with protein extracts from cells lacking the hnRNP A1. We also show that a specific hepatotoxic inducer of the Cyp2a5 gene, pyrazole, increases the cytoplasmic levels of hnRNP A1 in vivo. Finally, we show that hnRNP A1 can be overexpressed in mouse primary hepatocytes, leading to an accumulation of the CYP2A5 mRNA. Collectively, these results indicate that the hnRNP A1 is an important regulator of the Cyp2a5 gene. PMID- 11901219 TI - Involvement of activator protein-1 in transcriptional regulation of the human mu opioid receptor gene. AB - mu-Opioid receptors mediate such opioid effects as analgesia, euphoria, and immunomodulation. Gene expression of mu-opioid receptors can be modulated by various substances, including cytokines, hormones, and drugs. Some of these stimuli (e.g., IL-1beta and cocaine) have been shown to activate members of the AP-1 transcription factor family. In addition, transcription of the mu-opioid receptor gene is induced by the phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA), an activator of protein kinase C, which in turn is an activator of AP-1 transcription factors. This indicates that signaling pathways involving protein kinase C and activator protein 1 (AP-1) transcription factors are important for the specific expression pattern of the mu-opioid receptor gene. In this report, we show that TPA activates AP-1 as well as the transcription factor nuclear factor kappaB (NFkappaB) in the mu-opioid receptor expressing neuroblastoma cell line SH SY5Y. In transfection experiments performed in these cells, both factors trans-activate expression of reporter gene constructs containing the human mu-opioid receptor gene promoter. By excluding the effects of TPA on NFkappaB with the specific NFkappaB inhibitor sulfasalazine, AP-1 regulatory elements were localized. Two AP-1 elements, which differ in one nucleotide each from the classic AP-1 binding site, were delineated to positions 2388 and -1434 of the promoter. Independent of their orientation, these elements conferred TPA responsiveness on the heterologous thymidine kinase promoter. AP-1 binding to these elements was confirmed using electrophoretic mobility shift and immunoshift assays. PMID- 11901221 TI - Regulation of extracellular signal-regulated kinase cascades by alpha- and beta isoforms of the human thromboxane A(2) receptor. AB - Thromboxane A(2) (TXA(2)) stimulates mitogenic growth of vascular smooth muscle. In humans, TXA(2) signals through two TXA(2) receptor (TP) isoforms, termed TPalpha and TPbeta. To investigate the mechanism of TXA(2)-mediated mitogenesis, regulation of extracellular signal-regulated kinase (ERK) signaling was examined in human embryonic kidney 293 cells stably overexpressing the individual TP isoforms. The TXA(2) mimetic 9,11-dideoxy-9alpha,11alpha-methano epoxy prostaglandin F(2alpha) (U46619) elicited concentration- and time-dependent activation of ERK1 and -2 through both TPs with maximal TPalpha- and TPbeta mediated ERK activation observed after 10 and 5 min, respectively. U46619 mediated ERK activation was inhibited by the TP antagonist [1S-[1alpha,2beta-(5Z) 3beta,4alpha-]]-7-[3-[[2-(phenylamino)carbonyl]hydrazine] methyl]-7-oxabicyclo[ 2,2,1-]hept-2yl]-5-heptenoic acid (SQ29,548), and by the mitogen-activated protein kinase kinase inhibitor 2'-amino-3'-methoxyflavone (PD 98059). Although ERK activation through TPalpha was dependent on 2-[1-(dimethylaminopropyl)-1H indol-3-yl]-3-(1H-indol-3-yl)-maleimide (GF 109203X)-sensitive protein kinase (PK) Cs, ERK activation through TPbeta was only partially dependent on PKCs. ERK activation through both TPalpha and TPbeta was dependent on PKA and phosphoinositide 3-kinase (PI3K) class 1(A), but not class 1(B), and was modulated by Harvey-Ras, A-Raf, c-Raf, and Rap1B/B-Raf and also involved transactivation of the epidermal growth factor receptor. Additionally, PKB/Akt was activated through TPalpha and TPbeta in a PI3K-dependent manner. In conclusion, we have defined the key components of TXA(2)-mediated ERK signaling and have established that both TPalpha and TPbeta are involved. TXA(2)-mediated ERK activation through the TPs is a complex event involving PKC-, PKA-, and PI3K dependent mechanisms in addition to transactivation of the EGF receptor. TPalpha and TPbeta mediate ERK activation through similar mechanisms, although the time frame for maximal ERK activation and PKC dependence differs. PMID- 11901220 TI - Regulation of dopamine D(1) receptor trafficking by protein kinase A-dependent phosphorylation. AB - The aim of this study was to use pharmacological inhibition of protein kinase A and mutation of potential protein kinase A phosphorylation sites to determine the role of protein kinase A-catalyzed phosphorylation of the dopamine D(1) receptor in agonist-stimulated desensitization and internalization of the receptor. To facilitate purification and imaging of the D(1) receptor, we attached a polyhistidine tag to the amino terminus and enhanced green fluorescent protein to the carboxyl terminus of the receptor (D(1)-EGFP). D(1)-EGFP was similar to the untagged D(1) receptor in terms of affinity for agonist and antagonist ligands, coupling to G proteins, and stimulation of cyclic AMP accumulation. D(1)-EGFP and two mutants in which either Thr268 or Ser380 was replaced with Ala were stably expressed in NS20Y neuroblastoma cells. Pretreatment with the protein kinase A inhibitor H-89 (N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide) or substitution of Ala for Thr268 reduced agonist-stimulated phosphorylation of the receptor and resulted in diminished trafficking of the receptor to the perinuclear region of the cell. Substitution of Ala for Thr268 had no effect, however, on agonist-induced receptor sequestration or desensitization of cyclic AMP accumulation. Substitution of Ala for Ser380 had no effect on D(1) receptor phosphorylation, sequestration, desensitization, or trafficking to the perinuclear region. We conclude that protein kinase A-dependent phosphorylation of the D(1) receptor on Thr268 regulates a late step in the sorting of the receptor to the perinuclear region of the cell, but that phosphorylation of Thr268 is not required for receptor sequestration or maximal desensitization of cyclic AMP accumulation. PMID- 11901222 TI - Induction of rat organic anion transporting polypeptide 2 by pregnenolone-16alpha carbonitrile is via interaction with pregnane X receptor. AB - The rat organic anion transporting polypeptide 2 (oatp2; Slc21a5) is a liver transporter that mediates the uptake of a variety of structurally diverse compounds, and has a high affinity for cardiac glycosides. Treatment of rats with pregnenolone-16alpha-carbonitrile (PCN), a ligand for the rodent pregnane X receptor (PXR), significantly enhances the rat oatp2 gene expression. To understand the molecular mechanism of oatp2 induction by PCN, rat oatp2 gene was cloned. The rat oatp2 gene consists of 16 exons; alternative splicing of the second noncoding exon gives rise to the two published rat oatp2 cDNAs. Approximately 8700 base pairs (bp) of the 5'-flanking region of the rat oatp2 gene were linked to the luciferase reporter gene and used in transient transfection assays in H4IIE cells. Treatment of PCN induced the expression of the reporter gene in a dose-dependent manner. Four potential PXR response elements (PXREs) were identified in the 5'-flanking region of the rat oatp2 gene. One element (DR3-1) is located approximately -5000 bp with three more (DR3-2, DR3 3, and DR3-4) clustered at about -8000 bp. Results from electrophoretic mobility shift assays showed that the PXR-retinoid X receptor alpha heterodimer binds to the DR3-2 with the highest affinity, to the DR3-4 and DR3-1 with a lower affinity, and weakly or not at all to the DR3-3. Furthermore, a series of partial deletions of the 5'-flanking region illustrated that both the proximal and distal clusters of PXREs are required for maximal induction of rat oatp2 by PCN. In conclusion, these data elucidate the molecular mechanism by which PCN treatment induces rat oatp2 gene expression. In addition, this study identifies rat oatp2 as a direct PXR-targeted gene and further supports the hypothesis that activation of PXR affects a network of genes that is involved in either metabolism or transport of drugs, steroids, and bile acids. PMID- 11901223 TI - Cloning of CYP2J2 gene and identification of functional polymorphisms. AB - CYP2J2 is abundant in cardiovascular tissue and active in the metabolism of arachidonic acid to eicosanoids that possess potent anti-inflammatory, vasodilatory, and fibrinolytic properties. We cloned and sequenced the entire CYP2J2 gene (approximately 40.3 kb), which contains nine exons and eight introns. We then sequenced the CYP2J2 exons and intron-exon boundaries in 72 healthy persons representing African, Asian, and European/white populations as part of the National Institutes of Health/National Institute of Environmental Health Sciences Environmental Genome Single Nucleotide Polymorphism Program. A variety of polymorphisms were found, four of which resulted in coding changes (Arg158Cys, Ile192Asn, Asp342Asn, and Asn404Tyr). A fifth variant (Thr143Ala) was identified by screening a human heart cDNA library. All five variant cDNAs of CYP2J2 were generated by site-directed mutagenesis and expressed in Sf9 insect cells by using a baculovirus system. The recombinant wild-type and variant CYP2J2 proteins immunoreacted with peptide-based antibodies to CYP2J2 and displayed typical cytochrome P450 (P450) CO-difference spectra; however, the Asn404Tyr and Ile192Asn variants also had prominent spectral peaks at 420 nm. The ability of these variants to metabolize arachidonic acid and linoleic acid was compared with that of wild-type CYP2J2. Three variants (Asn404Tyr, Arg158Cys, and Thr143Ala) showed significantly reduced metabolism of both arachidonic acid and linoleic acid. The Ile192Asn variant showed significantly reduced activity toward arachidonic acid only. The Asp342Asn variant showed similar metabolism to wild type CYP2J2 for both endogenous substrates. Based on these data, we conclude that allelic variants of the human CYP2J2 gene exist and that some of these variants result in a P450 protein that has reduced catalytic function. Insofar as CYP2J2 products have effects in the cardiovascular system, we speculate that these variants may be functionally relevant. PMID- 11901224 TI - Carriers involved in targeting the cytostatic bile acid-cisplatin derivatives cis diammine-chloro-cholylglycinate-platinum(II) and cis-diammine-bisursodeoxycholate platinum(II) toward liver cells. AB - Molecular bases for targeting bile acid-cisplatin derivatives Bamet-R2 [cis diammine-chloro-cholylglycinate-platinum(II)] and Bamet-UD2 [cis-diammine bisursodeoxycholate-platinum(II)] toward liver cells were investigated. Carriers for bile acids [human Na(+)-taurocholate cotransporting polypeptide (NTCP)], organic anions [organic anion transporting polypeptide (OATP)], and organic cations [organic cation transporter (OCT)] were expressed in Xenopus laevis oocytes (XO) and Chinese hamster ovary (CHO) cells. Drug uptake was measured by flameless atomic absorption of platinum. Rat Oatp1- or rat Ntcp-transfected CHO cells were able to take up Bamets, but not cisplatin, severalfold more efficiently than wild-type cells. This uptake was enhanced by butyrate-induced expression of both carriers. Uptake of both Bamets by Ntcp-transfected CHO cells was stimulated by extracellular sodium. The amount of Bamets, but not cisplatin, taken up by XO was enhanced when expressing OATP-A, OATP-C, NTCP, OCT1, or OCT2, a nonhepatic OCT isoform used for comparative purposes. Bamet uptake by XO was inhibited by known substrates of these carriers (glycocholate for NTCP and OATP C, ouabain for OATP-A, and quinine for OCT1 and OCT2). Drug uptake versus substrate concentration revealed saturation kinetics (K(m) was in the 8-58 microM range), with the following order of efficiency of transport (V(max)/K(m)) for Bamet-R2: OATP-C > OCT2 > OATP-A > NTCP > OCT1; and the following order of efficiency of transport for Bamet-UD2: OATP-C > OCT2 > OATP-A > OCT1 > NTCP. Increasing the generation of cationic forms of Bamets by incubation in the absence of chloride increased drug uptake by OATP-A, OCT1, and OCT2 but reduced that achieved by NTCP and OATP-C. These results suggest a role for carriers of organic anions and cations in Bamet-R2 and Bamet-UD2 uptake, which may determine their ability to accumulate in liver tumor cells and/or be taken up and efficiently excreted by hepatocytes. PMID- 11901225 TI - Tracazolate reveals a novel type of allosteric interaction with recombinant gamma aminobutyric acid(A) receptors. AB - Tracazolate, a pyrazolopyridine, is an anxiolytic known to interact with gamma aminobutyric acid (GABA)(A) receptors, adenosine receptors, and phosphodiesterases. Its anxiolytic effect is thought to be via its interaction with GABA(A) receptors. We now report the first detailed pharmacological study examining the effects of tracazolate on a range of recombinant GABA(A) receptors expressed in Xenopus laevis oocytes. Replacement of the gamma2s subunit within the alpha1beta3gamma2s receptor with the epsilon subunit caused a dramatic change in the functional response to tracazolate from potentiation to inhibition. The gamma2s subunit was not critical for potentiation because alpha1beta3 receptors were also potentiated by tracazolate. gamma2/epsilon chimeras revealed a critical N-terminal domain between amino acids 206 and 230 of gamma2, governing the nature of this response. Replacement of the beta3 subunit with the beta1 subunit within alpha1beta3gamma2s and alpha1beta3epsilon receptors also revealed selectivity of tracazolate for beta3-containing receptors, determined by asparagine at position 265 within transmembrane 2. Replacement of gamma2s with gamma1 or gamma3 revealed a profile intermediate to that of alpha1beta1epsilon and alpha1beta1gamma2s. alpha1beta1delta receptors were also potentiated by tracazolate; however, the maximum potentiation of the EC(20) was much greater than on alpha1beta1gamma2. Concentration-response curves to GABA in the presence of tracazolate for alpha1beta1epsilon and alpha1beta1gamma2s revealed a concentration-related decrease in maximum current amplitude, but a leftward shift in the EC(50) only on alpha1beta1gamma2. Like alpha1beta1gamma2s, GABA concentration-response curves on alpha1beta1delta receptors were shifted to the left with increased maximum responses. Tracazolate has a unique pharmacological profile on recombinant GABA(A) receptors: its potency (EC(50)) is influenced by the nature of the beta subunit; but more importantly, its intrinsic efficacy, potentiation, or inhibition is determined by the nature of the third subunit (gamma1-3, delta, or epsilon) within the receptor complex. PMID- 11901226 TI - Delayed formation of hydrogen peroxide mediates the lethal response evoked by peroxynitrite in U937 cells. AB - The toxicity paradigm used in the present study involves exposure of U937 cells to a concentration of authentic peroxynitrite, leading to a rapid necrotic response mediated by mitochondrial permeability transition. We found that addition of catalase after treatment with peroxynitrite specifically prevents the loss of mitochondrial membrane potential and the ensuing lethal response. The protective effects of catalase were mimicked by the cocktail glutathione peroxidase/reduced glutathione. A defensive role of intracellular catalase was implied by experiments showing that catalase-depleted cells are hypersensitive to peroxynitrite and that cells with an increased catalase content, selected for their resistance to H(2)O(2), are cross-resistant to peroxynitrite. Further experiments demonstrated that H(2)O(2) formation takes place after peroxynitrite exposure. Various approaches using inhibitors of the mitochondrial respiratory chain as well as respiration-deficient cells revealed that the oxidant is produced upon dismutation of superoxides generated at the level of complex III. Interestingly, respiration-deficient cells were found to be resistant to peroxynitrite toxicity, and all those treatments increasing formation of H(2)O(2) produced a parallel increase in toxicity. In conclusion, the results presented in this study indicate that peroxynitrite-induced impairment of electron transport from cytochrome b to cytochrome c1 leads to delayed formation of hydrogen peroxide, which plays a pivotal role in the ensuing necrotic response. PMID- 11901227 TI - Down-regulation of DNA topoisomerase IIalpha in human colorectal carcinoma cells resistant to a protoberberine alkaloid, berberrubine. AB - Berberrubine, a protoberberine alkaloid that exhibits antitumor activity in animal models, has been identified as a specific poison of DNA topoisomerase II in vitro. To better understand the mechanisms of cellular response to berberrubine, human colorectal carcinoma cells (AMC5) were selected for resistance to berberrubine. The resulting cell line (AMC5/B1) was 5.3-fold resistant to berberrubine in the absence of MDR1 overexpression. The AMC5/B1 line was cross-resistant to topoisomerase II-targeted drugs but showed no cross resistance to other antitumor drugs. The patterns of cross-resistance to various drugs led us to examine the cellular contents of topoisomerase II. Topoisomerase II activity was approximately 2.8-fold lower in AMC5/B1 cells compared with parental cells. The AMC5/B1 line contained approximately 5-fold decrease in topoisomerase IIalpha protein level and approximately 2.5-fold decrease in topoisomerase IIalpha mRNA level. A comparison of the degradation kinetics of topoisomerase IIalpha mRNA demonstrated that there was no difference in mRNA stability between the two cell lines. Furthermore, the activity of topoisomerase IIalpha promoter in AMC5/B1 cells was about 25% of that in AMC5 parental cells when transient transfection experiments were performed with the promoter luciferase reporter gene. These results indicate that down-regulation of topoisomerase IIalpha in AMC5/B1 cells occurs at the transcriptional level. Nucleotide sequencing of the topoisomerase IIalpha promoter regions revealed no mutations in AMC5/B1 cells. In summary, resistance to berberrubine in AMC5 cells is associated with decreased level of catalytically active topoisomerase IIalpha, suggesting that topoisomerase IIalpha is the cellular target of berberrubine in vivo. PMID- 11901228 TI - Dopamine transporter mutants with cocaine resistance and normal dopamine uptake provide targets for cocaine antagonism. AB - Cocaine's blockade of dopamine reuptake by brain dopamine transporters (DAT) is a central feature of current understanding of cocaine reward and addiction. Empirical screening of small-molecule chemical libraries has thus far failed to provide successful cocaine blockers that allow dopamine reuptake in the presence of cocaine and provide cocaine "antagonism". We have approached this problem by assessing expression, dopamine uptake, and cocaine analog affinities of 56 DAT mutants in residues located in or near transmembrane domains likely to play significant roles in cocaine recognition and dopamine uptake. A phenylalanine-to alanine mutant in putative DAT transmembrane domain 3, F154A, retains normal dopamine uptake, lowers cocaine affinity 10-fold, and reduces cocaine stereospecificity. Such mutants provide windows into DAT structures that could serve as targets for selective cocaine blockers and document how combined strategies of mutagenesis and small molecule screening may improve our abilities to identify and design compounds with such selective properties. PMID- 11901229 TI - Cytochrome P450 3A conjugation to ubiquitin in a process distinct from classical ubiquitination pathway. AB - We characterize a novel microsome system that forms high-molecular-mass (HMM) CYP3A, CYP2E1, and ubiquitin conjugates, but does not alter CYP4A or most other microsomal proteins. The formation of the HMM bands was observed in hepatic microsomes isolated from rats treated 1 week or more with high doses (50 mg/kg/day) of nicardipine, clotrimazole, or pregnenolone 16alpha-carbonitrile, but not microsomes from control, dexamethasone-, nifedipine-, or diltiazem treated rats. Extensive washing of the microsomes to remove loosely attached proteins or cytosolic contaminants did not prevent the conjugation reaction. In contrast to prototypical ubiquitination pathways, this reaction did not require addition of ubiquitin, ATP, Mg(2+), or cytosol. Addition of cytosol did result in the degradation of the HMM CYP3A bands in a process that was not blocked by proteasome inhibitors. Immunoprecipitated CYP3A contained HMM ubiquitin. Even so, mass spectrometric analysis of tryptic peptides indicated that the HMM CYP3A was in molar excess to ubiquitin, suggesting that the formation of the HMM CYP3A may have resulted from conjugation to itself or a diffuse pool of ubiquitinated proteins already present in the microsomes. Addition of CYP3A substrates inhibited the formation of the HMM CYP3A and the cytosol-dependent degradation of HMM CYP3A. These results suggest that after extended periods of elevated CYP3A expression, microsomal factors are induced that catalyze the formation of HMM CYP3A conjugates that contain ubiquitin. This conjugation reaction, however, seems to be distinct from the classical ubiquitination pathway but may be related to the substrate-dependent stabilization of CYP3A observed in vivo. PMID- 11901230 TI - The batrachotoxin receptor on the voltage-gated sodium channel is guarded by the channel activation gate. AB - Batrachotoxin (BTX), from South American frogs of the genus Phyllobates, irreversibly activates voltage-gated sodium channels. Previous work demonstrated that a phenylalanine residue approximately halfway through pore-lining transmembrane segment IVS6 is a critical determinant of channel sensitivity to BTX. In this study, we introduced a series of mutations at this site in the Na(v)1.3 sodium channel, expressed wild-type and mutant channels in Xenopus laevis oocytes, and examined their sensitivity to BTX using voltage clamp recording. We found that substitution of either alanine or isoleucine strongly reduced channel sensitivity to toxin, whereas cysteine, tyrosine, or tryptophan decreased toxin action only modestly. These data suggest an electrostatic ligand receptor interaction at this site, possibly involving a charged tertiary amine on BTX. We then used a mutant channel (mutant F1710C) with intermediate toxin sensitivity to examine the properties of the toxin-receptor reaction in more detail. In contrast to wild-type channels, which bind BTX almost irreversibly, toxin dissociation from mutant channels was rapid, but only when the channels were open, not when they were closed. These data suggest the closed activation gate trapped bound toxin. Although BTX dissociation required channel activation, it was, paradoxically, slowed by strong membrane depolarization, suggesting additional state-dependent and/or electrostatic influences on the toxin binding reaction. We propose that BTX moves to and from its receptor through the cytoplasmic end of the open ion-conducting pore, in a manner similar to that of quaternary local anesthetics like QX314. PMID- 11901231 TI - Determinants of 4-aminopyridine sensitivity in a human brain kv1.4 k(+) channel: phenylalanine substitutions in leucine heptad repeat region stabilize channel closed state. AB - The biophysical and pharmacological effects of individual phenylalanine-for leucine (Phe-for-Leu) substitutions in the leucine heptad repeat region located at the cytosolic surface of the channel pore, on whole-cell K(+) currents, were studied in cloned and mutated human brain Kvl.4 K(+) channels (hKvl.4) transiently transfected into HeLa cells. Although L2 and L5 are not considered part of the 4-aminopyridine (4-AP) binding site, unlike the L4 heptad leucine, Phe substitutions at L2 (L464) or L5 (L485) increase 4-AP sensitivity by 400 fold, as seen previously in the L4F mutant channel. Greater depolarizing shifts manifest in the voltage dependence of activation and inactivation in L2F (20 mV) and L5F (30 mV) than in L4F (10 mV) relative to hKv1.4. L1F (L457) and L3F (L471) increase 4-AP sensitivity by 8- and 150-fold, respectively, and produce depolarizing shifts in activation of approximately 5 mV without affecting inactivation. The apparent free energy differences of 4-AP binding in each mutant suggest enhanced drug-channel interactions (L2F > or = L4F > or = L5F > L3F > L1F). Deactivation kinetics are accelerated in L2F (11-fold), L5F (8-fold), L1F (5-fold), and L3F (2-fold), at -50 mV. All Phe-for-heptad-Leu substitutions produce gating changes suggesting variable stabilization of the channel closed state conformation, with L1F, L2F, and L5F exhibiting the strongest correlations between altered gating and increased 4-AP sensitivity. If 4-AP blocks the open channel by promoting closure of the activation gate (recent Armstrong-Loboda model), then changes in the leucine heptad repeat that stabilize the channel closed state may contribute to increased 4-AP sensitivity by amplifying the mechanism of 4-AP block. PMID- 11901232 TI - Effects of the anticonvulsant retigabine on cultured cortical neurons: changes in electroresponsive properties and synaptic transmission. AB - The whole-cell patch-clamp technique was used to examine the effects of retigabine, a novel anticonvulsant drug, on the electroresponsive properties of individual neurons as well as on neurotransmission between monosynaptically connected pairs of cultured mouse cortical neurons. Consistent with its known action on potassium channels, retigabine significantly hyperpolarized the resting membrane potentials of the neurons, decreased input resistance, and decreased the number of action potentials generated by direct current injection. In addition, retigabine potentiated inhibitory postsynaptic currents (IPSCs) mediated by activation of gamma-aminobutyric acid(A) (GABA(A)) receptors. IPSC peak amplitude, 90-to-10% decay time, weighted decay time constant, slow decay time constant, and, consequently, the total charge transfer were all significantly enhanced by retigabine in a dose-dependent manner. This effect was limited to IPSCs; retigabine had no significant effect on excitatory postsynaptic currents (EPSCs) mediated by activation of non-N-methyl-D-aspartate ionotropic glutamate receptors. A form of short-term presynaptic plasticity, paired-pulse depression, was not altered by retigabine, suggesting that its effect on IPSCs is primarily postsynaptic. Consistent with the hypothesis that retigabine increases inhibitory neurotransmission via a direct action on the GABA(A) receptor, the peak amplitudes, 90-to-10% decay times, and total charge transfer of spontaneous miniature IPSCs were also significantly increased. Therefore, retigabine potently reduces excitability in neural circuits via a synergistic combination of mechanisms. PMID- 11901233 TI - Functional expression of a novel ginsenoside Rf binding protein from rat brain mRNA in Xenopus laevis oocytes. AB - We have shown that ginsenoside Rf (Rf) regulates voltage-dependent Ca(2+) channels through pertussis toxin (PTX)-sensitive G proteins in rat sensory neurons. These results suggest that Rf can act through a novel G protein-linked receptor in the nervous system. In the present study, we further examined the effect of Rf on G protein-coupled inwardly rectifying K(+) (GIRK) channels after coexpression with size-fractionated rat brain mRNA and GIRK1 and GIRK4 (GIRK1/4) channel cRNAs in Xenopus laevis oocytes using two-electrode voltage-clamp techniques. We found that Rf activated GIRK channel in a dose-dependent and reversible manner after coexpression with subfractions of rat brain mRNA and GIRK1/4 channel cRNAs. This Rf-evoked current was blocked by Ba(2+), a potassium channel blocker. The size of rat brain mRNA responding to Rf was about 6 to 7 kilobases. However, Rf did not evoke GIRK current after injection with this subfraction of rat brain mRNA or GIRK1/4 channel cRNAs alone. Other ginsenosides, such as Rb(1) and Rg(1), evoked only slight induction of GIRK currents after coexpression with the subfraction of rat brain mRNA and GIRK1/4 channel cRNAs. Acetylcholine and serotonin almost did not induce GIRK currents after coexpression with the subfraction of rat brain mRNA and GIRK1/4 channel cRNAs. Rf evoked GIRK currents were not altered by PTX pretreatment but were suppressed by intracellularly injected guanosine-5'-(2-O-thio) diphosphate, a nonhydrolyzable GDP analog. These results indicate that Rf activates GIRK channel through an unidentified G protein-coupled receptor in rat brain and that this receptor can be cloned by the expression method demonstrated here. PMID- 11901234 TI - The virally encoded fungal toxin KP4 specifically blocks L-type voltage-gated calcium channels. AB - KP4 is a virally encoded fungal toxin secreted by the P4 killer strain of Ustilago maydis. Previous studies demonstrated that this toxin inhibits growth of the target fungal cells by blocking calcium uptake rather than forming channels, as had been suggested previously. Unexpectedly, this toxin was also shown to inhibit voltage-gated calcium channel activity in mammalian cells. We used whole cell patch-clamp techniques to further characterize this activity against mammalian cells. KP4 is shown to specifically block L-type calcium channels with weak voltage dependence to the block. Because KP4 activity is abrogated by calcium, KP4 probably binds competitively with calcium to the channel exterior. Finally, it is shown that chemical reagents that modify lysine residues reduce KP4 activity in both patch-clamp experiments on mammalian cells and in fungal killing assays. Because the only lysine residue is K42, this residue seems to be crucial for both mammalian and fungal channel activity. Our results defining the type of mammalian channel affected by this fungal toxin further support our contention that KP4 inhibits fungal growth by blocking transmembrane calcium flux through fungal calcium channels, and imply a high degree of structural homology between these fungal and mammalian calcium channels. PMID- 11901235 TI - On the history of lacunes, etat crible, and the white matter lesions of vascular dementia. AB - The history of lesions associated with vascular dementia (17th to 19th century) is reviewed. Recognition of ischemic and hemorrhagic stroke types dates back to the 17th century; however, at that time a third type ('cerebral congestion') emerged as the most common form of apoplexy. This entity vanished as arterial hypertension became established with the introduction of the sphygmomanometer (1905). Before the 19th century, apoplexy was considered a uniformly fatal disease, although Willis first recognized post-stroke dementia in 1672. Dechambre (1838) first reported 'lacunes' in stroke survivors with small cerebral softenings. Durand-Fardel (1842) described interstitial atrophy of the brain (leukoaraiosis) and etat crible (cribriform state) reflecting chronic cerebral congestion. In 1894, Alzheimer and Binswanger identified 'arteriosclerotic brain atrophy,' a form of vascular dementia characterized by 'miliary apoplexies' (lacunes). Also in 1894, Binswanger described the disease that now bears his name. In 1901, Pierre Marie coined the name etat lacunaire (lacunar state) for the clinical syndrome of elderly patients with multiple lacunes. PMID- 11901236 TI - Pathophysiology of age-related cerebral white matter changes. AB - The pathogenesis of age-related cerebral white matter changes (WMC) is still a matter of investigation. Alterations of deep small vessels, such as arteriolosclerosis, are considered to play a central role in the development of WMC. Stenosis or occlusion of small vessels may cause sudden or more chronic ischemia resulting in small areas of necrosis (lacunar infarction) or diffuse alterations consistent with the definition of white matter incomplete infarct. Moreover, the arteriolosclerotic changes may cause loss of autoregulation in the deep white matter and consequent cerebral blood flow fluctuations in response to changes of systemic blood pressure. Both these types of mechanisms may be particularly harmful because the blood supply of the white matter is of the terminal type with scarce anastomoses. Other pathophysiological hypotheses on the origin of WMC have been raised, and they can probably be considered as complementary to the ischemic one. The small vessel alterations could lead to damage of the blood-brain barrier and chronic leakage of fluid and macromolecules in the white matter. The increased interstitial fluid concentration in abnormal white matter may be also a consequence of arterial hypertensive bouts. Genetically determined factors could play an important role in the development of WMC, and at least one disease (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy - CADASIL) characterized by severe leukoencephalopathy exists with a determined genetic origin. It is possible that other genetic factors contribute, by interaction with conventional risk factors, to the development of white matter injury in nonfamilial cases. PMID- 11901237 TI - Neuropathology of white matter lesions in vascular cognitive impairment. AB - The white matter is an important locus for tissue damage in vascular cognitive impairment and white matter lesions often dominate over gray matter changes. The spectrum of ischemic white matter lesions histopathologically represents focal and diffuse lesions, the most common form being the combination of both, in varying proportions. In the combined pathology, the diffuse lesion represents a gradient zone of damage towards surrounding normal tissue and may hold over 200 times the volume of an identified focal lesion, the lacunar infarct. Pathogenetically, the focal lesion results from the acute reaction to regional ischemia, while the diffuse white matter lesion represents the adjustment to altered perfusional and physiological conditions within the tissue. PMID- 11901238 TI - Evolution of white matter lesions. AB - A 3-year follow-up of 273 participants (mean age 60 years) of the Austrian Stroke Prevention Study provides first information on the rate, clinical predictors and cognitive consequences of MRI white matter lesions (WML) in elderly individuals without neuropsychiatric disease. Lesion progression was found in 17.9% of individuals over a time period of 3 years. Diastolic blood pressure and early confluent or confluent white matter hyperintensities at baseline were the only significant predictors of white matter hyperintensity progression. Genetic association studies in the setting of the Austrian Stroke Prevention Study provide first evidence for genetic susceptibility factors for progression of WML. We observed associations with the paraoxonase Leu-->Met 54 polymorphism and with the M235T polymorphism of the angiotensinogen gene. Lesion progression had no influence on the course of neuropsychologic test performance over the observational period, but the statistical power of this analysis was low. PMID- 11901240 TI - CT and MRI rating of white matter lesions. AB - Rating scales play an important role in the evaluation of computed tomography (CT) or magnetic resonance-detected white matter lesions (WML). Unfortunately, this type of visual semiquantitative assessment is not yet an optimal tool because commonly agreed concepts regarding its use are lacking. To generate a discussion platform for further improvement of CT and MRI rating, we will provide some basic definitions, summarize the advantages and disadvantages of scoring schemes and review current efforts towards the improvement of this tool. Future research will have to concentrate on deepening our understanding of the histopathologic substrates of WML and on strategies to document their progression. PMID- 11901239 TI - Imaging of white matter lesions. AB - Magnetic resonance imaging (MRI) is very sensitive for the detection of white matter lesions (WML), which occur even in normal ageing. Intrinsic WML should be separated from physiological changes in the ageing brain, such as periventricular caps and bands, and from dilated Virchow-Robin spaces. Genuine WML are best seen with T2-weighted sequences such as long TR dual-echo spin-echo or FLAIR (fluid attenuated inversion recovery); the latter has the advantage of easily separating WML from CSF-like lesions. Abnormal T2 signal in MRI is not specific, and can accompany any change in tissue composition. In the work-up of WML in small vessel disease, magnetic resonance angiography can be used to rule out (concomitant) large vessel disease, and diffusion-weighted MRI to identify new ischaemic lesions (amidst pre-existing old WML). The differential diagnosis of WML includes hereditary leukodystrophies and acquired disorders. The leukodystrophies that can present in adult age include metachromatic leukodystrophy, globoid cell leukodystrophy, adrenomyeloneuropathy, mitochondrial disorders, vanishing white matter, and cerebrotendinous xanthomatosis. These metabolic disorders typically present with symmetrical abnormalities that can be very diffuse, often with involvement of brainstem and cerebellum. Only the mitochondrial disorders tend to be more asymmetric and frequently involve the grey matter preferentially. Among the acquired white matter disorders, hypoxic-ischaemic causes are by far the most prevalent and without further clinical clues there is no need to even consider the next most common disorder, i.e. multiple sclerosis (MS). Among the nonischaemic disorders, MS is far more common than vasculitis, infection, intoxication and trauma. While vasculitis can mimic small vessel disease, MS has distinctive features with preferential involvement of the subcortical U-fibres, the corpus callosum, temporal lobes and the brainstem/cerebellum. Spinal cord lesions are very common in MS, but do not occur in normal ageing nor in small vessel disease. PMID- 11901241 TI - CADASIL: a monogenic condition causing stroke and subcortical vascular dementia. AB - Mutations in Notch3 are the cause of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), an inherited small vessel disease leading to subcortical strokes and vascular dementia. The phenotypic presentation is variable but remarkable for a high frequency of migraine with aura. Magnetic resonance images show a microangiopathic pattern of lesions. Prominent involvement of the temporopolar white matter and involvement of the temporopolar arcuate fibers are conspicuous findings seen in many patients. The underlying angiopathy is characterized by a unique type of ultrastructural basal lamina deposits and by degeneration of vascular smooth muscle cells which are the major source of Notch3 expression. In line with these findings there is evidence for a functional impairment of vascular smooth muscle cells. CADASIL has opened a new perspective in studying basic mechanisms of vessel wall degeneration and ischemic tissue damage related to small vessel disease. PMID- 11901242 TI - Cerebral amyloid angiopathy and vessel dysfunction. AB - Cerebral amyloid angiopathy (CAA), defined by deposition of the beta-amyloid peptide in medium and small cortical and meningeal vessels, is a well-recognized cause of hemorrhagic stroke. This paper reviews the accumulating evidence supporting an additional role for CAA in producing vessel dysfunction, reduced cerebral blood flow and ischemia. Ischemic lesions are characteristic of several hereditary CAA syndromes, including a recently described mutation of the amyloid precursor protein associated with dementia (but not hemorrhagic stroke) in an Iowa family. Ischemic lesions are seen in some sporadic CAA patients as well, and recent data from transgenic mice suggest potential mechanisms by which beta amyloid may alter vessel physiology. Future studies will seek to define the clinical importance of vascular beta-amyloid as a potential target for drug therapy in dementia. PMID- 11901244 TI - Cognition and white matter lesions. AB - Although it is recognized that ischemic stroke is a potent risk factor for vascular dementia, the influence of white matter lesions (WML) on cognitive function is less clear. In community-based MRI studies that have administered mental status tests to subjects who were free of clinically evident neurologic disease, a weak relationship between WML and generalized cognitive function has been reported. In studies that have administered neuropsychological test batteries, a stronger and more specific association has been recognized between WML and deficits in executive function, most likely due to the involvement of frontal-subcortical pathways. Cognitive deficits may be related to the total volume of the WML, with a threshold perhaps needing to be surpassed before such deficits are evident, but it is likely that the location of the WML also plays a role, with that threshold varying in association with the distribution of the lesions. Potential confounders of the results of previous studies include small, strategically located subcortical infarctions that may be masked by more extensive WML and other comorbid neurologic disorders, particularly Alzheimer's disease. Future studies should be prospective, utilize standardized methods for structural and functional brain imaging, and administer comprehensive neuropsychological assessments in order to more rigorously investigate the relationship between evolving WML and declining cognitive functions. PMID- 11901243 TI - Small vessel disease and Alzheimer's dementia: pathological considerations. AB - Current evidence suggests that the neuropathology of Alzheimer type of dementia comprises more than amyloid plaques and neurofibrillary tangles. At least a third of Alzheimer disease (AD) cases may exhibit significant cerebrovascular pathology, which constitutes distinct small vessel disease (SVD). Cerebral amyloid angiopathy, microvascular degeneration affecting the cerebral endothelium and smooth muscle cells, basal lamina alterations, hyalinosis and fibrosis are often evident in AD. These changes may be accompanied by perivascular denervation that is causal in the cognitive decline of AD. Amyloid beta protein may cause degeneration of both the larger perforating arterial vessels as well as cerebral capillaries, which represent the blood-brain barrier. In addition, macro- and microinfarctions, haemorrhages, lacunes and ischaemic white matter changes are also present in AD. The development of SVD in late-onset AD may engage an interaction of perivascular mediators as well as circulation-derived factors that perturb the brain vasculature. Peripheral vascular disease such as long-standing hypertension, atrial fibrillation, coronary or carotid artery disease and diabetes could further modify the cerebral circulation such that a sustained hypoperfusion or oligaemia is impacted upon the ageing brain. PMID- 11901245 TI - Subcortical vascular dementia. AB - Vascular dementia (VaD) incorporates different vascular mechanisms and changes in the brain, and has different causes and clinical manifestations. Current criteria for VaD select an aetiologically and clinically heterogeneous group and this definitional heterogeneity has affected clinical trial results. Focus on a more homogeneous group, such as that with subcortical (ischaemic) VaD, could be an alternative in clinical drug trials. This subtype incorporates two small vessel clinical entities - 'Binswanger's disease' and 'the lacunar state'. It comprises small vessel disease as the primary vascular aetiology, lacunar infarct(s) and ischaemic white matter lesions as the primary type of brain lesions, subcortical location as the primary location of lesions, and the subcortical clinical syndrome as the primary clinical manifestation. Subcortical VaD is expected to show a more predictable clinical picture, natural history, outcome and treatment responses. PMID- 11901246 TI - New approaches to clinical trials in vascular dementia: memantine in small vessel disease. AB - Although criteria for the diagnosis of vascular dementia (VaD) are established, the diagnostic concept is still controversial and there is no regulatory guidance for clinical drug development. Clinical trials in VaD present a number of pitfalls and challenges and, so far, no compound has received regulatory approval for this indication. The methodological issues of clinical VaD trials are discussed using the development of memantine for this indication as an example. In a pooled analysis of two placebo-controlled trials with the NMDA-antagonist memantine in VaD, the cognitive benefit by memantine treatment was more pronounced for patients with 'small vessel disease' than for those with other neuroradiological findings at baseline. In a subgroup of patients with 'large vessel disease' or macrolesions, there was less cognitive decline among the placebo patients. It may therefore be helpful to use predefined diagnostic subcategories in clinical studies in this indication. The findings further suggest that stroke or multiple infarctions may not be the primary reason for cognitive decline in VaD patients. PMID- 11901249 TI - Penetration of ravuconazole, a new triazole antifungal, into rat tissues. AB - Ravuconazole (BMS 207147, ER-30346) is a long-lasting triazole antifungal agent active against a broad spectrum of fungal pathogens including non-albicans Candida, Aspergillus, Cryptococcus and key dermatophytic fungi. The penetration of ravuconazole into rat tissues was examined. Fifty-five 7-week-old specific pathogen free female rats were used in this study. Plasma, lung and uterus tissue of rats were taken at 1, 2, 4, 8, 12, 16, 24, 32, 48, 60, and 72 h (n = 5) after oral administration of 10 mg/kg of ravuconazole. The quantitative assays of ravuconazole by HPLC after the extraction with diethylether were conducted for each tissue sample homogenate. tmax, t1/2, and Cmax of ravuconazole is 8 h, 16.9 h and 1.68 microg/ml, respectively. The concentrations of ravuconazole in rat uterus and lung tissues were 2-to 6 times higher than the corresponding blood concentrations. The ratio of plasma to lung levels of ravuconazole was superior to the published data of other azoles. Considering its antifungal spectrum, ravuconazole would thus be a good candidate for treatment of deep-seated fungal infections caused by Candida, Aspergillus and Cryptococcus. PMID- 11901250 TI - Selection of Klebsiella pneumoniae mutants with high-level cefotaxime resistance during growth in serum containing therapeutic concentrations of cefotaxime. AB - BACKGROUND: In a previous investigation of the genetic characterization of extended-spectrum beta-lactamases (ESBLs) in Klebsiella pneumoniae from Zagreb, Croatia, 20 strains were found to produce SHV-2 beta-lactamase. Those strains displayed varying degrees of beta-lactam resistance and a wide range of beta lactamase activity. We concluded that more resistant isolates were hyperproducers of SHV-2 beta-lactamase. METHODS: In this investigation, we tried to develop hyperproducing variants from 8 low-level SHV-2 beta-lactamase-producing Klebsiella strains by subculturing them in serum containing therapeutic concentrations of cefotaxime (CTX). RESULTS: In most cases, there was a moderate increase in CTX resistance (twofold to threefold), except in one strain which displayed a 16-fold increase in the minimum inhibitory concentration (MIC) of CTX after incubation in the serum. That strain showed a marked increase in enzyme activity as well. The strains with a moderate increase in CTX MIC did not produce more enzyme after exposure to the serum, except for one strain which had a threefold rise in beta-lactamase activity after exposure to serum. CONCLUSIONS: In this investigation, it was established that the mutants with high-level CTX resistance developed very quickly in the biological fluids containing therapeutic concentrations of CTX. It is reasonable to expect that a similar process occurs in patients infected with an ESBL-producing K. pneumoniae strain during antibiotic treatment. Since most of the high-level CTX-resistant mutants did not have a marked rise in beta-lactamase activity after exposure to serum, it is possible that the elevated resistance was due to some other mechanism, such as reduced penicillin-binding protein affinity, changes in outer membrane proteins or efflux by multidrug efflux pumps. PMID- 11901251 TI - MEN 10700, a new penem antibiotic: in vitro antibacterial activity on clinical isolates. AB - MEN 10700 is a new broad-spectrum penem, currently in preclinical development. In the present study, the activity of MEN 10700 was compared to that of imipenem, meropenem, cefotaxime, ampicillin/sulbactam, amikacin and ciprofloxacin against 619 gram-positive and gram-negative bacterial strains, and to that of imipenem, meropenem, cefotaxime, ceftriaxone, ceftazidime, cefepime and ampicillin/sulbactam against 38 strains of ciprofloxacin-resistant Escherichia coli, and against 19 extended-spectrum-beta-lactamase (ESBL)-producing strains of the KES group. MEN 10700 was highly active against most gram-positive and gram negative strains, and overall demonstrated comparable activity to imipenem and meropenem. Methicillin-susceptible Staphylococcus aureus and methicillin susceptible Staphylococcus epidermidis were highly susceptible to MEN 10700, which was the most potent among the antibiotics tested. MEN 10700 was less potent than the carbapenem antibiotics on Morganella morganii, Serratia marcescens and Acinetobacter spp. Ciprofloxacin-resistant E. coli were uniformly susceptible to MEN 10700, imipenem and meropenem, with MIC90 values in the range of < or = 0.12 0.25 mg/l, while showing much lower susceptibility to the other antibiotics tested, including the fourth-generation cephalosporin cefepime. This feature was even more evident in ESBL-producing strains of the KES group, with an MIC90 of 1- 2 mg/l for MEN 10700, imipenem and meropenem, and a MIC90 of 16-64 mg/l for the other antibiotics tested, including cefepime. PMID- 11901252 TI - Comparison of the Sensititre YeastOne colorimetric microdilution panel and the NCCLS broth microdilution method for antifungal susceptibility testing against Candida species. AB - We evaluated the commercially prepared Sensititre YeastOne colorimetric antifungal panel to determine the susceptibility of 170 Candida spp isolates to amphotericin B, fluconazole, itraconazole, and flucytosine. The NCCLS reference microdilution method (M27-A document) was used as reference method. The YeastOne panel was performed according to the manufacturer's instructions. For the colorimetric method, MICs were determined at 24 h of incubation. MICs for the NCCLS reference method were read at 48 h of incubation. The overall agreement within +/-2 dilutions by both methods was calculated against the four antifungal agents. This agreement was 92.9, 68.2, 77.6 and 80% for amphotericin B, fluconazole, itraconazole, and flucytosine, respectively. Thirteen isolates (7.6%) showed very major discrepancies for fluconazole and 12 (7%) for itraconazole. We found that the reading of MIC with the YeastOne panel was somewhat easier than the reading of reference MIC, although the determination of endpoint was sometimes difficult, especially for azoles, because the trailing effect appeared in a high percentage of isolates. PMID- 11901253 TI - Evaluation of the new VITEK 2 system for determination of the susceptibility of clinical isolates of Streptococcus pneumoniae. AB - BACKGROUND: The VITEK 2 is a new version of the automated system for organism identification and susceptibility testing. One of the differences between this system and its predecessor, VITEK, is the ability to perform rapid susceptibility testing of Streptococcus pneumoniae. This study compares the results of susceptibility testing of S. pneumoniae using the VITEK 2 system and a commercial microbroth dilution method, Sensititre. METHODS: A group of 214 clinical isolates of S. pneumoniae were selected to include isolates with previously documented penicillin resistance. The antimicrobial agents tested were benzylpenicillin, cefotaxime, erythromycin, trimethoprim/sulfamethoxazole, tetracycline, chloramphenicol, imipenem and vancomycin. RESULTS: The best agreement was achieved with vancomycin (100%), erythromycin (95.8%) and tetracycline (95.8%). The lowest level of agreement was found with benzylpenicillin (88.6%) and cefotaxime (90.1%). We observed rates of 12.3 and 15.7% for minor errors with penicillin and cefotaxime, respectively, and 1 very major error for cefotaxime. CONCLUSION: The VITEK 2 allows rapid determination of the antimicrobial susceptibility of S. pneumoniae and demonstrated a good degree of agreement with the Sensititre method for most of the antimicrobials tested. PMID- 11901254 TI - Investigation of various antibiotic combinations using the E-Test method in multiresistant Pseudomonas aeruginosa strains. AB - BACKGROUND: Antibiotic combinations are frequently used in order to obtain wide spectrum effects in the treatment of serious infections such as septicemia and endocarditis, and also to produce an in vivo effect against strains which are defined as resistant to the known inhibiting or fatal dose of one antibiotic. The synergistic effects of combinations such as aminoglycoside + beta-lactam, aminoglycoside + quinolone and quinolone + beta-lactam on Pseudomonas aeruginosa have been revealed in different studies. The multiple resistance rate of nosocomial P. aeruginosa strains isolated from intensive care units (ICUs) has been reported as high in many studies. METHODS: In this study, the effects of various combinations of antibiotics (aminoglycoside + beta-lactam and aminoglycoside + quinolone) against 101 multiresistant P. aeruginosa strains which were isolated from the ICUs of three different hospitals in Istanbul were investigated using the E-test method. The combinations for which the highest synergistic effects were determined by the E-test method were also tested with the checkerboard method, i.e. in addition to the E-test method, in 19 of a total of 23 strains. RESULTS: When the synergistic results which were obtained with the combinations of aminoglycoside + beta-lactam were compared with those of the aminoglycoside + quinolone combinations, they were determined to be higher for the two aminoglycosides gentamicin (GM) and tobramycin (TM). We determined the synergistic rates to be 23, 21, 19, 18, 16, 14, 10 and 10% for GM + ceftriaxone (TX), GM + piperacillin (PP), GM + ceftazidime (TZ), TM + PP, TM + TX, TM + TZ, GM + ciprofloxacin (CI) and TM + CI, respectively. The GM + TX combination - for which the highest synergistic effects were determined with the E-test stripes - was also determined as synergistic with the checkerboard method in 19 of a total of 23 strains (23%), and the agreement rate between the two methods was 100% (kappa > 0.7). The highest synergistic effects against strains which were sensitive to both of the antibiotics which constitute the combinations were found for the GM + TX combination, as 50%, whereas for strains which were resistant to both of the antibiotics, this was found for the TM + PP combination, also as 50%. CONCLUSIONS: We consider that the minimal inhibitory concentration values of antibiotics are not sufficient alone in order to constitute a combination for multiresistant strains and it would be advisable to begin a treatment by applying a combination study. The E-test method has been evaluated as a good alternative for combination investigations because of its ease both of application and evaluation and also for its good agreement with the standard checkerboard method. PMID- 11901255 TI - Cytotoxic effects of diazenes on tumor cells in vitro. AB - BACKGROUND: We have shown previously that diazenecarboxamides (diazenes) were cytotoxic for several tumor cell lines. Their target seems to be the intracellular glutathione (GSH). To improve the solubility and biological activity of these drugs, new compounds have been synthesized. In the present study we examined the cytotoxic effect of six new diazenes: BR-25, UP-39, UP-11, JK-1024, RL-514 and RL-625. METHODS: The cytotoxicity of diazenes was tested on nine human tumor cell lines: laryngeal carcinoma HEp2 cells, cervical carcinoma HeLa cells, mammary carcinoma MCF-7 cells, breast adenocarcinoma SK-BR-3 cells, and glioblastoma A1235 cells and their four drug-resistant cell lines. Cytotoxicity was determined using the colorimetric MTT assay. The intracellular GSH content (following the treatment with diazenes) was examined spectrophotometrically in human cervical carcinoma cells by the procedure developed by Tietze. RESULTS: Results show that diazene UP-39 was mostly efficient, significantly reducing the cell survival of all nine cell lines examined, including four drug-resistant cell lines. Diazenes UP-11, RL-514 and BR 25 given in the highest concentrations decreased the survival of some examined cell lines, but to less than 50%. Diazenes RL-625 and JK-1024 had no influence on the cell survival. None of the examined diazenes significantly influenced the intracellular level of GSH. CONCLUSION: Our results suggest that diazene UP-39 may be a promising new drug for the treatment of tumors and encourage further research on this compound. PMID- 11901257 TI - Ceftriaxone versus other cephalosporins for perioperative antibiotic prophylaxis: a meta-analysis of 43 randomized controlled trials. AB - The efficacy of ceftriaxone versus other cephalosporins in the perioperative prophylaxis of surgical wound, urinary tract and respiratory tract infections was compared in a meta-analysis of randomized controlled trials published between 1986 and 1996, identified from the Medline, Embase, SIGLE, ROPU, DHSS-Data and Medikat Cologne databases. Studies were grouped by type of infection, operative specialty, wound classification, study quality and other factors, and assessed for relative risk (RR). Forty-three studies with a total of 13,482 patients met our inclusion criteria. RR for surgical wound infection (n = 13,303 patients) was 30% lower in the ceftriaxone versus control groups [98.3% confidence interval (CI): 0.55-0.89; p = 0.0002]. In urinary tract infections (n = 8,865 patients), the primary analysis of all studies showed marked superiority for ceftriaxone (RR: 0.53; 98.3% CI: 0.43-0.67) but not in studies with CDC-defined infections (RR: 0.63; 98.3% CI: 0.36-1.12). In both types of infection, ceftriaxone was superior in contaminated operations. The data showed no advantage for ceftriaxone in other operations. In respiratory tract infections (n = 9,567 patients), there was no significant difference: the RR was 0.81 (98.3% CI: 0.61-1.09; p = 0.04). PMID- 11901258 TI - Susceptibility of 57 bloodstream and urinary isolates of Candida species from a single children's university hospital to 6 antifungals. PMID- 11901256 TI - Protection effects of Taurine supplementation against cisplatin-induced nephrotoxicity in rats. AB - BACKGROUND: Taurine, which is the major intracellular free beta-amino acid, is known to be an antioxidant and a membrane-stabilizing agent. This study was designed to investigate the protective role of taurine supplementation against cisplatin-induced nephrotoxicity. METHODS: Male Wistar rats were divided into six groups and treated as follows: (1) saline-treated control drinking tap water, (2) saline-treated plus taurine-supplemented (1.5% taurine in the drinking water), (3) saline-treated plus taurine-depleted (3% beta-alanine in the drinking water), (4) cisplatin-treated, CDDP 6 mg/kg intraperitoneally, (5) taurine-supplemented plus CDDP-treated and (6) taurine-depleted plus CDDP-treated. Rats were sacrificed 7 days after CDDP treatment, and serum as well as kidneys were isolated and analyzed. RESULTS: CDDP-treated rats showed increased kidney weight as a percentage of total body weight, serum creatinine and BUN levels and decreased serum albumin and calcium levels. Also, CDDP treatment resulted in a depletion of kidney GSH content, a reduction in the kidney glutathione peroxidase (GSH-Px) activity and increased kidney MDA production level. Taurine supplementation attenuated CDDP-induced nephrotoxicity which was manifested by jeopardizing the elevation in serum creatinine and BUN levels and the reduction in serum albumin and calcium levels. Moreover, taurine supplementation restored kidney GSH content and GSH-Px activity and reduced platinum accumulation and MDA production levels in the kidney tissue following CDDP treatment. Histopathological examination of the kidney of CDDP-treated rats revealed tubular atrophy, tubular necrosis and desquamation of renal tubular cells. However, taurine supplementation protected against CDDP-induced histopathological changes. CONCLUSIONS: The data suggest that taurine supplementation effectively attenuates the accumulation of platinum within kidney tissue and counteracts the deleterious effect of CDDP on the renal tubular function. PMID- 11901260 TI - A study of menses-related changes to the larynx in singers with voice abuse. AB - Fifteen vocally abusive and 15 healthy female classical singers were examined before and outside menses by means of laryngoscopy, laryngostroboscopy and electroglottography. Investigation before menses revealed more extensive alteration of the larynx in vocally abusive singers than in healthy ones. Subsequent investigation, after the inappropriate use of their voice had been eliminated, indicated decreased changes to the larynx. It was concluded that premenstrual hyperchanges were a result of combined abuse- and menses-related influence on the larynx, and that laryngeal examination before menstruation could be a useful test for the presence of physiologically correct singing. PMID- 11901261 TI - Aerodynamics of esophageal voice production with and without a Groningen voice prosthesis. AB - OBJECTIVE: Aerodynamic aspects of esophageal voice production in laryngectomees have been studied to clarify and compare the physiology of injection (IE) and button-assisted (TE) esophageal voice. METHODS: Simultaneous measurements of intratracheal, sub- and suprapseudoglottic pressure, transpseudoglottic flow and sound pressure level (SPL) in laryngectomees were carried out. Efficiency of substitute voice production and pressure loss over the prosthesis were calculated. Relationships between the variables were computed by means of regression analysis, and Pearson's product-moment correlations. RESULTS: (1) TE speakers use a higher subpseudoglottic pressure and a higher mean airflow rate during phonation than IE speakers; (2) in TE and IE speakers SPL values are about the same; (3) in assessing voice performance of TE speakers intratracheal air pressure is the most important aerodynamic variable. CONCLUSION: The aerodynamic model of button-assisted esophageal voice production presented here provides a basis for future studies in this field. PMID- 11901262 TI - [Influence of voice onset on the perceptual analysis of dysphonia]. AB - The perceptual analysis of voice for 40 dysphonic and normal subjects was studied according to the GRBAS method and with the consensus of three experts. In order to evaluate the effects of voice onset on the evaluation by the jury, we presented two sample materials for each subject: a sustained /alpha/ and the same sustained /alpha/ for which voice onset was suppressed. In a blind test, the two sample materials were presented in the same session in a random way. The jury did not notice the sustained /alpha/ for which voice onset was suppressed.The influence of voice onset on the jury was based on the percentage of identical answers for each material. Our results show that the influence of voice onset is correlated with the dysphonic level. Voice onset would be of major importance for intermediate dysphonia (G1, G2), whereas it seems not to exert any influence in the case of normal voices (G0) or severe dysphonia (G3). PMID- 11901263 TI - Changes during long-term management of locked-in syndrome: a case report. AB - This paper describes various approaches of treatment to a male client suffering from the locked-in syndrome following closed head injury. During the course of therapy, which started more than 5 years after onset and lasted for as much as 11 years, the client progressed from the so-called classical to the incomplete locked-in state. The different approaches as well as the outcomes are presented in detail, followed by a discussion about interdisciplinary issues, duration of treatment and the impact of a patient's personality on the course and goals of the treatment as well as on the mode of communication. PMID- 11901264 TI - The relevance of stroboscopy in functional dysphonias(1). AB - OBJECTIVES: Functional dysphonias are disorders of the voice characterized by sound and efficiency disturbances of the voice without any organic changes of structures being detectable. At present, functional dysphonias are generally subclassified into hyper- and hypofunctional dysphonias in clinical practice. STUDY DESIGN: The study was designed for a critical evaluation of the relevance of stroboscopy to the diagnostics and classification of functional dysphonias. METHODS: 45 patients were examined (27 hyperfunctional, 15 hypofunctional and 3 mixed type) using videostroboscopy. Several stroboscopic parameters were taken into consideration. Three geometrical and three time-dependent parameters were first analyzed in a uni- and multidimensional way, then cluster analyses were performed. RESULTS: We could not confirm the clinical subdivision into hyper- and hypofunctional dysphonias as based on anamnestic data, perceptual evaluation of voice sound, voice profile measurements and videostroboscopy. Quantitative measurements of selected parameters did not correlate with qualitative subjective stroboscopic assessment. In addition to this, it was not possible to identify separate clusters of stroboscopic findings. CONCLUSIONS: The results do not deny the clinical relevance of stroboscopy to the diagnostics of functional dysphonias as a very useful tool to exclude organic lesions. However, a reliable subclassification into different types of functional dysphonias was not possible. PMID- 11901265 TI - GENEHUNTER: your 'one-stop shop' for statistical genetic analysis? AB - The past decade has brought a proliferation of statistical genetic (linkage) analysis techniques, incorporating new methodology and/or improvement of existing methodology in gene mapping, specifically targeted towards the localization of genes underlying complex disorders. Most of these techniques have been implemented in user-friendly programs and made freely available to the genetics community. Although certain packages may be more 'popular' than others, a common question asked by genetic researchers is 'which program is best for me?'. To help researchers answer this question, the following software review aims to summarize the main advantages and disadvantages of the popular GENEHUNTER package. PMID- 11901266 TI - Single-nucleotide variant in multiple copies of a deleted in azoospermia (DAZ) sequence - a human Y chromosome quantitative polymorphism. AB - The evolution of the deleted in azoospermia (DAZ) gene family supports prevalent theories on the origin and development of sex chromosomes and sexual dimorphism. The ancestral DAZL gene in human chromosome 3 is known to be involved in germline development of both males and females. The available phylogenetic data suggest that some time after the divergence of the New World and Old World monkey lineages, the DAZL gene, which is found in all mammals, was copied to the Y chromosome of an ancestor to the Old World monkeys, but not New World monkeys. In modern man, the Y-linked DAZ gene complex is located on the distal part of the q arm. It is thought that after being copied to the Y chromosome, and after the divergence of the human and great ape lineages, the DAZ gene in the former underwent internal rearrangements. This included tandem duplications as well as a T > C transition altering an MboI restriction enzyme site in a duplicated sequence. In this study, we report on the ratios of MboI-/MboI+ variant sequences in individuals from seven worldwide human populations (Basque, Benin, Egypt, Formosa, Kungurtug, Oman and Rwanda) in the DAZ complex. The ratio of PCR MboI- and MboI+ amplicons can be used to characterize individuals and populations. Our results show a nonrandom distribution of MboI-/MboI+ sequence ratios in all populations examined, as well as significant differences in ratios between populations when compared pairwise. The multiple ratios imply that there have been more than one recent reorganization events at this locus. Considering the dynamic nature of this locus and its involvement in male fertility, we investigated the extent and distribution of this polymorphism. PMID- 11901267 TI - Frequency Distribution and Haplotypic Heterogeneity of beta(E)-Globin Gene among Eight Minority Groups of Northeast Thailand. AB - The frequencies of hemoglobin E and beta(E)-globin gene haplotypes were determined in eight minority groups living in the northeastern part of Thailand. A total of 478 samples of eight minority groups, namely Soui, Thai Khmer, So, Yor, Phuthai, Thai Puan, Thai Loei and Thai Dam, were examined. High prevalences of hemoglobin E (>50%) were observed in Soui, Thai Khmer, So, Yor and Phuthai inhabiting the region near Cambodia and Laos. Thai Puan, Thai Loei, Thai Dam and native Thai living in the same geographical area had prevalences of 42.6, 35.9, 21.4 and 27.9%, respectively. A prevalence of 9.5% was found among the Thai with Chinese background living in the same area. Beta-globin gene haplotypes analysis demonstrated that most of the beta(E)-globin genes in these Thai populations were associated with two haplotypes: (- + - + + + -) and (+ - - - - + -) on chromosomes with framework 2 variety. Some beta(E)-globin genes in Soui and Thai Khmer groups were associated with the framework 3 chromosome. Genetic distances based on the beta( )-globin gene haplotypes between minority groups revealed that Soui and Thai Khmer were closely related to each other. This finding has a valuable implication for study of the origin and spread of hemoglobin E in the region. PMID- 11901268 TI - HLODs, trait models, and ascertainment: implications of admixture for parameter estimation and linkage detection. AB - Maximizing the homogeneity lod is known to be an appropriate procedure for estimating parameters of the trait model in an approximately 'ascertainment assumption free' (AAF) manner. We have investigated whether this same property also holds for the heterogeneity lod (HLOD). We show that, when the genetic models at linked and unlinked loci differ, HLODs are not AAF, and maximizing the HLOD yields parameter estimates that are for all practical purposes meaningless; indeed, the admixture parameter alpha does not even measure the proportion of linked families within the sample, as is commonly supposed. In spite of this, our results confirm a large body of evidence supporting the use of HLODs as robust tools for linkage detection, and suggest further that maximizing the HLOD over both alpha and parameters of the trait model can improve accuracy in estimation of the recombination fraction theta;. These findings have important implications for the optimal handling of nuisance parameters in linkage analysis, particularly when evaluating the evidence for or against linkage based on multiple independent heterogeneous sets of data. PMID- 11901269 TI - Faster haplotype frequency estimation using unrelated subjects. AB - Linkage disequilibrium (LD) between tightly linked loci provides fine mapping information of disease-predisposing allelic variants. The most common method of LD analysis involves unrelated cases and controls. We have previously proposed model-free and permutation tests for diseases with unknown mode of inheritance that can be applied to several highly polymorphic loci. However, performing such analyses remained computer intensive. In this report we propose a speed-up of both the gene-counting procedure and the permutation procedure. We demonstrate the improved method with an analysis of schizophrenia and human leucocyte antigen markers, and an analysis of alcoholism and mitochondrial aldehyde dehydrogenase markers. Our implementation also allows the rapid calculation of permutation based LD measures and related statistics. PMID- 11901270 TI - Mutation analysis in a patient with succinic semialdehyde dehydrogenase deficiency: a compound heterozygote with 103-121del and 1460T > A of the ALDH5A1 gene. AB - We saw a 17-month-old boy with moderate psychomotor retardation, and enzymatically diagnosed succinic semialdehyde dehydrogenase (SSADH) deficiency. After extracting mRNA and genomic DNA from his cultured lymphoblasts, we analyzed the entire coding region of the ALDH5A1 gene using reverse transcription polymerase chain reaction (RT-PCR) and genomic PCR followed by sequencing. He was demonstrated to be a compound heterozygote with two novel mutations (103-121 del and 1460T>A). The former leads to a frameshift and premature termination, and the latter is a missense mutation, V487E. Both mutations were also detected in the genomic DNA. Taken together with previous mutation reports, genetic heterogeneity was suspected for SSADH deficiency, and may account for the wide range of its phenotype. PMID- 11901271 TI - The linkage information content value of polymorphism genetic markers in model free linkage analysis. AB - Guo and Elston [Hum Hered 1999;49:112-118] developed a linkage information content (LIC) value to measure the informativeness of a marker for identity-by descent (IBD) sharing status of relative pairs. LIC values were derived for five types of relative pairs: full sib, half sib, grandparent-grandchild, first cousin and avuncular. In this paper, we give corrected LIC values for full sib, grandparent-grandchild, first cousin and avuncular pairs, and indicate the availability of a computer program to calculate them. PMID- 11901273 TI - Hypothesis testing for data from different family study designs. AB - A test statistic that is valid for data collected according to a particular type of family study design is not necessarily valid when applied to data obtained from a different type of family study design. When this can occur, a different test that usually is valid is developed for each type of family study design. However, investigators might find that their data come from two (or more) different family study designs, each requiring a different test, yet they want an overall conclusion, essentially a valid hypothesis test that is as powerful as possible. When the underlying genetic model is unknown, it is not clear how to proceed, as several alternative approaches might appear feasible. By using as an example the development of a test of association for data concerning affected singletons and their parents and affected sib pairs and their parents, it is shown that it may not be possible to develop a universally optimal approach without knowledge of the underlying genetic model. PMID- 11901272 TI - Analysis of CCR5Delta32 geographic distribution and its correlation with some climatic and geographic factors. AB - We studied the possible effects of climatic-geographic factors on the world distribution of the mutant allele for the chemokine receptor gene CCR5, which has a 32-bp deletion (CCR5Delta32) preventing cell invasion by the primary transmitting strain of HIV-1. New data on CCR5 polymorphisms in Russian, Ukrainian, and Moldavian populations are presented. All available data on CCR5Delta32 frequencies in the Old World (number of populations n = 77) were used for construction of a geographical gene map to analyze possible correlations between allele frequencies and eight climatic-geographic parameters. A strong positive correlation was found between the allele frequency and latitude (r = 0.72), a strong negative correlation with annual radiation balance (r = -0.66), and a weaker negative correlation with longitude (r = -0.34). Partial correlations were calculated excluding the influence of latitude. The negative correlation between the allele frequency and annual radiation balance decreased (r = -0.42), but remained large and significant. We propose that the existence of correlations between the cline of CCR5Delta32 frequencies and climatic-geographic parameters provides evidence for a possible effect of either natural environmental factors or large-scale population movements on the distribution of this allele. PMID- 11901274 TI - Impaired olfaction predicts cognitive decline in nondemented older adults. AB - In 1992, the twelve-item Brief Smell Identification Test((R)) and, in 1992 and 1996, a variety of measures of verbal learning and memory, executive control, and global function were administered to a total of 359 individuals (286 men and 73 women; mean age in 1992 74.3 years). Individuals with a history of stroke or impaired cognition at baseline were excluded from analyses. Impaired olfactory function (present or absent) was related to a greater 4.5-year decline on several indices of verbal memory, but not to a decline on measures of executive control or of global functioning after adjustment for baseline cognitive performance, age, education, gender, and history of smell difficulties. Olfactory loss remained associated with a decline in components of verbal memory, independently of the apolipoprotein E epsilon4 status. The predictive utility of impaired smell identification in older adults appears to be specific to a decline in components of verbal memory. PMID- 11901275 TI - Severe dysosmia is specifically associated with Alzheimer-like memory deficits in nondemented elderly retirees. AB - OBJECTIVES: To determine whether or not (1) impaired olfactory function is associated with impaired memory on neuropsychological testing in healthy retirees, and if so then (2) whether memory impairment is most consistent with a mesiotemporal rather than frontal system disorder. METHODS: 173 independent residents of a continuing care retirement community were studied. Subjects completed the University of Pennsylvania Smell Identification Test (UPSIT) and a battery of both general and specific cognitive measures that included the Mini Mental State Examination (MMSE) and the Executive Interview (EXIT25). Subjects were examined twice over 3 years. RESULTS: UPSIT performance was normal in 21% and in the 'anosmic' range in 25% of subjects. Anosmic UPSIT performance was associated with significantly worse performance on all cognitive tests. However, only short-term verbal memory was independently associated with UPSIT-defined anosmia. This association remained significant after adjusting for the other cognitive and sociodemographic variables. The memory deficits of anosmic subjects were qualitatively consistent with a cortical type (type 1) dementing illness such as Alzheimer's disease (AD). Over time, UPSIT-defined 'anosmic' cases suffered significantly greater declines on both the MMSE and the EXIT25, independently of baseline age, gender and MMSE score. CONCLUSIONS: Impaired odor identification in individuals without overt dementia is associated with an AD like memory impairment and an increased rate of cognitive decline. The comorbid association of these deficits is consistent with the known hierarchical spread of preclinical AD pathology and may be a specific indicator of future clinical AD dementia. PMID- 11901276 TI - Prevalence of dementia and apolipoprotein e genotype distribution in the elderly of buttapietra, verona province, Italy. AB - We investigated the prevalence of dementia and the apolipoprotein E (APOE) genotype distribution in the elderly of Buttapietra, a village near Verona, Italy. All residents over the age of 74 (n = 238), including those who were institutionalized, were studied using a direct-contact, single-phase design. The overall prevalence of dementia, clinically defined by DSM-III-R criteria, was 15.8 cases per 100 population, with age-specific figures increasing steeply with advancing age in both sexes. Alzheimer's disease (AD) was the most frequent dementing disorder (43%). APOE genotyping was determined after DNA amplification by restriction isotyping. We found that the epsilon4 allele and the epsilon3/epsilon4 genotype were associated with all types of dementia, although only the association of epsilon3/epsilon4 with AD reached statistical significance (odds ratio 4.5, 95% confidence interval 1.3-16.1). However, as reported in other Mediterranean countries, the frequency of the epsilon4 allele in our population was low (8.9%), suggesting that the population-attributable risk for AD, at least for elderly individuals (> or =75 years), could be small. PMID- 11901277 TI - A case-control analysis of nonsteroidal anti-inflammatory drugs and Alzheimer's disease: are they protective? AB - In many studies of nonsteroidal anti-inflammatory drugs (NSAIDs) and Alzheimer's disease (AD), the exposure to NSAIDs was concurrent with AD or based on self (or surrogate) report. We conducted a case-control analysis of the Quebec participants in the Canadian Study of Health and Aging who received a diagnosis of AD (cases) or were found to be cognitively unimpaired on screening (controls). Information on drug use was obtained from the Quebec Provincial Pharmaceutical Services Database. There was no significant difference in the proportion of cases and controls who had received any NSAID prescriptions in the 3 years prior to the onset of symptoms of dementia; amongst NSAID users, there was no difference in mean dose or duration. Our findings, using a measure of drug use prior to symptom onset and not subject to recall bias, do not support a protective effect for NSAIDs. PMID- 11901278 TI - Evaluation and correction for a 'training effect' in the cognitive assessment of older adults. AB - BACKGROUND: Repeated administration of psychometric instruments frequently results in a higher score on retesting, the so-called training effect. Yet, a training effect has been poorly considered in longitudinal studies of cognitive changes in older persons. METHODS: We investigated the presence, magnitude and potential adjustments for training effect in the older participants of the Systolic Hypertension in the Elderly Program (SHEP). SHEP evaluated the cognitive status effects of a diuretic-based treatment of isolated systolic hypertension versus placebo. Changes in the short Comprehensive Assessment and Referral Evaluation (short-CARE) questionnaire score, from baseline through 4 years of follow-up, were assessed in 4,718 participants. In this study, we used two regression techniques to adjust data for the training effect. RESULTS: In both study groups, a training effect was evident as a progressive improvement in the short-CARE score throughout year 1. Thereafter, cognitive scores tended to deteriorate, more in the placebo than in the active treatment group (p = 0.055). When follow-up scores were adjusted based upon baseline data, the difference between the study groups reached statistical significance (p = 0.019), but the apparent overall trend towards deterioration in cognitive score was no longer observed. Adjustment of baseline data preserved this apparent temporal course, but did not improve the discrimination between the two study groups (p = 0.165). CONCLUSIONS: In SHEP, repeated cognitive assessments were likely biased by a training effect which could be only partially corrected by statistical techniques. Studies of changes in the cognitive status of older persons should be designed appropriately to estimate and minimize the consequences of a training effect in follow-up data. PMID- 11901280 TI - Lifetime prevalence of Bell's palsy in rural Bolivia: a door-to-door survey. AB - We carried out a door-to-door survey in rural areas of the Cordillera Province, Bolivia, to determine the prevalence of the most common neurological diseases in a sample of about 10,000 inhabitants. A team of non-doctor health workers administered a standard screening instrument for neurological diseases, a slightly modified version of the World Health Organization protocol. All subjects found positive at the screening phase underwent a complete neurological examination. On screening, we found 1,130 positive subjects, of whom 909 were aged 15 years and above. After the neurological examination, we found 52 cases who had experienced Bell's palsy during their life in the population aged 15 years and above. The lifetime prevalence on November 1, 1994 was 11.1/1,000 (95% confidence interval 7.8-14.5) for the population aged 15 years and above. The prevalence was higher in women than in men (13.7 and 8.7/1,000, respectively) and increased with age, reaching a peak in the group aged 65 years or more (31.7/1,000). Only 3 cases (5.8%) had received medical therapy. PMID- 11901279 TI - Anticardiolipin antibodies and mortality in patients with ischemic stroke: a prospective follow-up study. AB - The purpose of the present prospective observational study was to assess whether or not the presence of anticardiolipin antibodies (aCL) in unselected first ischemic stroke patients is associated with adverse outcome. Consecutive patients (n = 300; mean age 64 years; 48% males) presenting with a first acute ischemic stroke were evaluated for IgG aCL and were systematically followed up. During a median follow-up of 21 months, 58 patients (19%) died. Mortality rates were higher in patients with aCL >20 IgG phospholipid units (GPL) [33 vs. 18%; relative risk (RR) 1.94, 95% confidence interval (CI) 1.02-3.67; p = 0.042] or >40 GPL (40 vs. 19%; RR 2.46, 95% CI 1.05-5.75; p = 0.037). Elevated aCL did not confer an increased risk during follow-up of a combined end point of stroke, myocardial infarction and vascular death or of nonfatal thrombo-occlusive events. Rates of malignancy detected during follow-up were higher among patients with aCL >20 GPL (19 vs. 5%, p = 0.007) and >40 GPL (27 vs. 6%, p = 0.01). The excess mortality associated with elevated aCL was eliminated after adjustment for age, cardiovascular risk factors and malignancy. These results demonstrate that aCL above 20-40 GPL among consecutive ischemic stroke patients is a marker of increased mortality during follow-up, but older age and higher rates of cardiovascular risk factors and malignancy detected during follow-up account for the higher mortality. PMID- 11901281 TI - Corneal allograft rejection: current understanding. 2. Clinical implications. PMID- 11901282 TI - Ultrasound biomicroscopy of residual vitreous base after vitreoretinal surgery. AB - Residual vitreous base after vitreoretinal surgery was evaluated by ultrasound biomicroscopy (UBM). Twenty aphakic and pseudoaphakic patients (20 eyes) undergoing surgery for different vitreoretinal diseases were evaluated by high frequency (50 MHz), high-resolution (50 microm) UBM, performed the day before surgery, weekly up to 1 month after surgery and then monthly. One week after surgery, the vitreous remnants were 'hardly visible' in 3 cases, 'visible' in 6 cases and 'highly visible' in 11. At the end of the follow-up (2.2 months), the 'hardly visible' cases increased to 6 and the 'visible' cases to 12, while the 'highly visible' cases decreased to 2. UBM demonstrated that vitreous base remnants were present in spite of accurate surgery; a spontaneous volume reduction of vitreous was observed during the follow-up. PMID- 11901283 TI - [Color duplex ultrasound of the temporal artery: replacement for biopsy in temporal arteritis]. AB - The diagnosis of temporal arteritis (TA) is generally confirmed by biopsy. To investigate the diagnostic accuracy of color duplex sonography (CDS), both temporal arteries of 20 patients with suspected TA were prospectively insonated prior to biopsy. Detection of >or=1 hypoechogenic perivascular halo was used as CDS criterion, a temporal artery biopsy and the criteria of the American College of Rheumatology (ACR) as references. The frequency of halo disappearance after 3 months of steroid therapy was also studied. CDS showed TA in 6, biopsy in 12 and ACR criteria in 15 patients. CDS sensitivity was 50 and 40%, and specificity 100%, using the biopsy and the ACR criteria, respectively. After 3 months of steroid treatment, 1 patient still showed halos. In conclusion, detection of halos confirms, whereas the absence of halos does not exclude the diagnosis of TA suggesting that ultrasound may replace biopsy in single patients with typical clinical signs and symptoms and a halo. PMID- 11901284 TI - Postural modifications of the oscillatory potentials of the electroretinogram in primary open-angle glaucoma. AB - The postural variations in the retinal microcirculation in glaucomatous patients were studied by evaluation of the oscillatory potentials (OPs) of the ERG. The OPs in scotopic adaptation were examined in subjects with primary open-angle glaucoma and in an age- and sex-matched normal control group in different body positions (seated, supine, anti-Trendelenburg, Trendelenburg). In the seated position, the difference of mean OP amplitude between the control group and the glaucomatous patients was highly significant (p < 0.001). In the normal subjects the OP amplitude in the Trendelenburg position was statistically lower with respect to the values obtained in all other positions (p < 0.05). In the glaucomatous patient group, the OP amplitude in the anti-Trendelenburg position was increased as compared to the other positions (p < 0.05). The study showed a reduction in amplitude of OPs in glaucomatous patients and their different behaviour in both groups with changes in body position. PMID- 11901285 TI - Comparisons of risk factors and visual field changes between juvenile-onset and late-onset primary open-angle glaucoma. AB - PURPOSE: To identify the differences in risk factors and visual field (VF) changes between juvenile primary open-angle glaucoma (JOAG) and late-onset chronic open-angle glaucoma (COAG). METHODS: The demographic and presenting clinical data of 27 JOAG and 30 COAG patients were retrospectively reviewed. Comparisons between the two groups were performed using Mann-Whitney U test, Wilcoxon signed-rank test, and Fisher's exact test. RESULTS: A family history of glaucoma (37%) and a history of steroid usage (14.8%) were identified in JOAG patients only. The JOAG patients had a longer axial length (p < 0.001) and more often a myopic refractive state (p < 0.001) than the COAG patients. Patients with COAG showed a deeper (p = 0.016) and a more extensive (p = 0.008) defect in the superior than in the inferior hemifield, as well as a deeper (p = 0.016) and a more extensive (p = 0.001) defect in the superior than in the inferior arcuate area, while JOAG patients showed symmetric VF defects between the superior and inferior hemifields. Purely diffuse VF depression is more common in JOAG than in COAG patients (p = 0.03). CONCLUSIONS: JOAG patients demonstrated more axial myopic changes than patients with COAG as well as a pattern of superior-inferior symmetric VF defects. Axial myopia may play a critical role in the pathogenesis of JOAG. PMID- 11901286 TI - Selection of scleral buckling for primary retinal detachment. AB - PURPOSE: Rhegmatogenous retinal detachment (RRD) may be caused by a flap tear or by an atrophic hole along the lattice degeneration. The aim of this study was to see whether different types of scleral buckling could achieve comparable reattachment rates in eyes with specific types of RRD. PATIENTS AND METHODS: 128 eyes with RRD were assigned to receive 1 of 3 buckling procedures according to the following guidelines: retinal detachments caused by flap tears were treated with radial segmental buckling; retinal detachments caused by atrophic holes with limited lattice degeneration were treated with circumferential segmental buckling, and retinal detachments caused by multiple breaks with extensive lattice degeneration were treated with encircling buckling. RESULTS: 56 eyes received radial segmental buckling, 36 eyes received circumferential segmental buckling, and 36 eyes received encircling buckling. The reattachment rates in these three groups were 83.9, 86.1, and 88.9%, respectively (no statistically significant difference). The visual outcomes were comparable in all groups. Younger age, an increased requirement for subretinal fluid drainage, longer operation time, and myopic shift were noted in the encircling group. CONCLUSIONS: Comparable reattachment rates could be achieved in all three groups according to our guidelines. Segmental buckling is appropriate for two thirds of RRD in this study and has fewer complications than encircling buckling. Every retinal detachment behaves differently and should be subjected to its optimal buckling procedure to achieve the best results and to avoid unnecessary operative complications. PMID- 11901287 TI - Impact of increased intraocular pressure on long-term corneal endothelial cell density after penetrating keratoplasty. AB - PURPOSE: Besides immunological graft rejection, persistently increased intraocular pressure (IOP) is among the most important causes for graft failure after penetrating keratoplasty (PK). The purpose of this study was to assess the longitudinal development of IOP after PK and to investigate possible correlations with corneal endothelial cell density. METHODS: This longitudinal prospective study included 209 eyes after PK with a complete follow-up at 3 months, 6 months, before first suture removal (16 +/- 5 months) and after complete suture removal (21 +/- 5 months). At each examination, IOP was measured by Goldmann applanation tonometry. Endothelial cell density was assessed by specular microscopy (EM 1100, Tomey). The indications for PK were 48% keratoconus, 34% Fuchs' dystrophy, 5.4% stromal dystrophies, 8.6% secondary bullous keratopathy and 4% corneal scars. An iridotomy was performed routinely during PK. The postoperative treatment with topical steroids was standardized. RESULTS: Preoperatively, the mean IOP was 13.6 +/- 2.9 mm Hg with increased IOP (>21 mm Hg) in 2.0% of eyes. After 3 months, the incidence of increased IOP (24.5 +/- 4.6 mm Hg) was highest (6.7%) and decreased thereafter to 3.0% after complete suture removal. No patients showed IOP higher than 30 mm Hg. A persistently increased IOP for more than 3 months was seen in 2% of patients. At 3 months postoperatively, the mean endothelial cell density was 1,977 +/- 496/mm(2) and did not decrease significantly (p > 0.05) until 6 months (1,771 +/- 507 cells/mm(2)). At the end of the follow-up period, the mean endothelial cell density was significantly reduced (1,347 +/- 501 cells/mm(2)). There was no significant correlation between IOP and mean endothelial cell density at any postoperative examination stage (p > 0.24). Patients with persistently increased IOP for more than 3 months did not have significantly different endothelial cell densities in comparison with those without increased IOP. CONCLUSION: From 6 months after PK, the incidence of increased IOP was not higher than the preoperative level. Mild to moderate temporary IOP elevations after PK do not seem to affect the endothelial cell density of the graft until complete suture removal. Further long-term studies are necessary to assess the clinical relevance of these observations. PMID- 11901289 TI - Effect of botulinum toxin A on facial wrinkle lines in Koreans. AB - Two kinds of botulinum toxin type A were clinically evaluated in rhytidectomy. Twenty Korean patients with facial wrinkles were fully assessed following treatments with random injections. The mean degree of wrinkles before the injections was 2.83 and the mean corrective effect was 70.0% at least 3 months afterward. The effect lasted less than 6 months in only 9 cases. The complications were tingling sensations in 3 cases (15.0%), temporary lid swelling in 5 cases (25.0%) and lagophthalmos in 3 cases (15.0%). No serious or permanent adverse effects were observed. Botulinum toxin type A rhytidectomy was a very effective method of removing various facial wrinkles although the treatment for complications and side effects will need to be considered. PMID- 11901288 TI - A comparison of the short-term hypotensive effects and side effects of unilateral brimonidine and apraclonidine in patients with elevated intraocular pressure. AB - PURPOSE: To compare the short-term ocular hypotensive efficacy and side effects of 0.2% brimonidine and 0.5% apraclonidine in patients with elevated intraocular pressure (IOP). METHODS: We performed a double-masked, placebo-controlled study to compare the efficacy of the application of 0.2% brimonidine and 0.5% apraclonidine for the effect of IOP, systemic blood pressure and heart rate in 20 newly diagnosed ocular hypertensive patients. Effects on the untreated fellow eye and ocular side effects were also determined. All measurements were performed 1, 2, 4, 6 and 8 h after the instillation of one drop. RESULTS: Brimonidine and apraclonidine significantly reduced IOP from baseline at all observation times. No significant difference was observed between the treatment groups. IOP decreased significantly in the untreated fellow eye in the brimonidine group at 4 , 6- and 8-hour checks and at 6-hour checks in the apraclonidine group when compared with placebo. Blood pressure and heart rate decreased significantly in the brimonidine group compared with placebo. Apraclonidine did not affect blood pressure or heart rate any differently than placebo. The pupil diameter and the interpalpebral fissure width significantly increased in the apraclonidine group, but not in the brimonidine group. There were no significant differences in the overall incidence of foreign body sensation, burning and stinging and dry mouth in the treatment groups. CONCLUSIONS: In the short-term, brimonidine was effective in reducing IOP in patients with elevated IOP and was equivalent in efficacy to apraclonidine. On the other hand, a significant change in blood pressure and heart rate was observed with brimonidine; there was no change at all in the apraclonidine group. PMID- 11901290 TI - Ocular firework injuries at New Year's eve. AB - PURPOSE: To prospectively study mechanisms and injury characteristics of ocular firework burns. METHODS: A prospective analysis of all patients with firework injuries attending the Department of Ophthalmology, University of Vienna, between 1994 and 2001. We looked for classes of fireworks and mechanisms of injuries. The number of diagnoses was established and their severity classified. RESULTS: During this period (8 years) we identified 116 eyes of 102 patients. 67/102 (66%) of all injuries were caused by class II and III fireworks. Patients ranged between 4 and 83 years. Ocular injuries occurred more frequently in males (69, 68%) and affected the right eye in 53 patients (52.5%). Ocular firework injuries to minors (under the age of 18 years) occurred in 48 (49%). The most common types of injury were skin and corneal erosions and abrasions (32/116, 28%). 11/116 eyes (10%) had severe injuries. 8 of 11 severe injuries (72%) occurred in minors (<18 years). 2 patients (1.7%) developed permanent blindness. In 32/116 patients (28%), the ocular trauma resulted in visual impairment, mainly due to corneal scars or retinal pathologies. In all patients, the severity index was 1.4 +/- 0.8. In minors the severity index was 1.6 +/- 1.0, in adults 1.2 +/- 0.5 (p = 0.013). CONCLUSION: Injuries in minors were significantly more severe than those in adults. Possible preventive measures include legislation, education of minors and eye protection. PMID- 11901291 TI - Effect of sympathetic denervation on rabbit choroidal blood flow. AB - In this study we demonstrate the existence of sympathetic innervation and compare the effect of unilateral or bilateral superior cervical sympathectomy on albino rabbit choroidal blood flow (CBF) during changes in perfusion pressure (PP). Forty albino rabbits weighing between 2.0 and 3.0 kg were randomly divided into three groups. The bilateral sympathectomy group (group S) included 10 rabbits (20 eyes) that received bilateral sympathectomy 1 week prior to the study. The unilateral sympathectomy group (group U) included 20 rabbits (20 eyes) that received unilateral sympathectomy 1 week prior to the study. Only the eyes ipsilateral to sympathectomy were utilized. The other 10 rabbits (20 eyes) served as controls (group N), each received the same procedure as the experimental groups except that the superior cervical ganglion (SCG) was preserved. The blood cell flux (PF), velocity (V), and concentration of moving blood cells (CMBC) were recorded simultaneously by means of a laser Doppler flowmeter (Perimed PF4001), while the intraocular pressure was increased linearly with a syringe pump. When the PP decreased steadily, the PF, V, and CMBC remained constant until PP <55 mm Hg, then decreased proportionally to the PP. When the PP decreased from 75 to 0 mm Hg, the PF, V and CMBC decreased from 100 to 6.87 +/- 0.97%, 8.44 +/- 0.92%, and 18.67 +/- 0.91% in group N, to 18.56 +/- 1.62%, 19.30 +/- 1.84%, and 38.02 +/ 9.10% in group U, and to 18.38 +/- 2.89%, 16.78 +/- 1.48%, and 34.58 +/- 4.42% in group S. The changes in PF, V, and CMBC were similar in groups S and U. Both group S and U had higher PF, V, and CMBC values than group N at comparable PPs below 55 mm Hg. These results indicate that the SCG plays a role in CBF regulation. Both unilateral and bilateral sympathectomy led to a higher PF in both groups S and U rabbits, indicating increased CBF while PP decreased gradually. This suggests that the rabbit choroid does not receive crossed innervation. The plateau response in each curve demonstrates the presence of autoregulation. This autoregulation was unchanged by either unilateral or bilateral sympathetic denervation. PMID- 11901292 TI - Optic neuropathy from folic acid deficiency without alcohol abuse. AB - A 47-year-old woman with a 2-month history of bilateral progressive visual loss was found to have a bilateral retrobulbar optic neuropathy. Her serum vitamin B(12) concentration and hemoglobin level were normal, but her serum folic acid concentration was decreased. The patient had a minimal alcohol intake and moderate tobacco use that had been unchanged for over 20 years; however, she had markedly altered her diet 4 years earlier in the setting of clinical depression. After treatment with oral folic acid and diet modification without change in her tobacco or alcohol use, the patient's visual function returned to normal. This case supports the role of folic acid deficiency as an important cause of some cases of nutritional optic neuropathy. PMID- 11901293 TI - Anterior ischemic optic neuropathy associated with idiopathic aldosteronism and hypertension. AB - A 49-year-old man suffered from bilateral anterior ischemic optic neuropathy almost simultaneously, and was diagnosed with idiopathic aldosteronism associated with hypertension. Because this patient had multiple organ disorders, multiple cerebral infarctions, and a mild loss of renal function, it was important to treat his primary disease. PMID- 11901294 TI - Adie's tonic pupil-induced angle-closure glaucoma. AB - A 52-year-old woman was diagnosed as having Adie's tonic pupil in her right eye. She reported few episodes of blurring of vision in her right eye in the recent few months. On one of the routine follow-up visits, right intraocular pressure (IOP) was 70 mm Hg and on gonioscopy the angle was closed 360 degrees. Medical treatment resulted in IOP reduction and laser iridotomy was then performed. This is, to our knowledge, the first description of intermittent angle-closure glaucoma attacks induced by a tonic pupil. The possibility of angle-closure glaucoma should be considered in patients with a tonic pupil, especially with symptoms of blurred vision or ocular pain. PMID- 11901295 TI - Activation of the metallothionein IIA promoter and other key stress response elements by ursodeoxycholate in HepG2 cells: relevance to the cytoprotective function of ursodeoxycholate. AB - Ursodeoxycholate, used to treat a variety of pathologies, has the ability to reverse cytotoxic and hepatotoxic conditions. We examined HepG2, a hepatic cell line, treated with increasing levels of ursodeoxycholate, for responses of a range of promoters/response elements responsive to DNA damage, heavy metal ions, protein denaturants, aromatic hydrocarbons, retinoids, changes in intracellular AMP levels, end endoplasmic reticulum stress. The metallothionein IIA promoter was the most highly activated by ursodeoxycholate. Since ursodeoxycholate protects against the cytotoxic effects of deoxycholate, our data, combined with observations made by others, implicate metallothionein IIA as being important in this protective pathway. PMID- 11901296 TI - Diminished inibitory action of ethanol on the contraction of gallbladder isolated from chronically ethanol-fed Guinea pigs. AB - Ethanol is known to decrease the gallbladder contractility. The purpose of this study was to clarify the mechanism of tolerance to the inhibitory action of ethanol on the gallbladder contractility. Male guinea pigs were fed ethanol (3%) or calorie-matched sucrose in the drinking water for 4 weeks. Then, the gallbladder was isolated, and its isometric tension was measured. The contractile responses to KCl, BAY K8644, histamine, and phorbol 12,13-dibutyrate in the normal medium were not different between the gallbladder strips from ethanol-fed and control guinea pigs. Ethanol at 25 mmol/l in vitro did not affect the contractile responses to KCl and BAY K8644 in the gallbladder strips from both ethanol-fed and control guinea pigs. On the other hand, ethanol at 25 mmol/l in vitro significantly inhibited the contractile responses to histamine and phorbol 12,13-dibutyrate in the gallbladder strips from the control guinea pigs, but it did not affect the contractile response to histamine and significantly augmented that to phorbol 12,13-dibutyrate in the strips from the ethanol-fed guinea pigs. Diphenhydramine, a selective H(1) receptor antagonist, abolished the histamine contraction in gallbladder strips from both control and ethanol-fed guinea pigs, while cimetidine, a selective H(2) receptor antagonist, did not affect histamine contraction, implying that histamine-induced contraction of guinea pig gallbladders is mediated only by H(1) receptors. Verapamil (1 micromol/l) completely inhibited the phorbol 12,13-dibutyrate-induced contraction of the strips from both ethanol-fed and control guinea pigs. The histamine-induced contraction was partly inhibited in the absence of Ca(2+) in the medium. In the gallbladder strips from both ethanol-fed and control guinea pigs, ethanol at 25 mmol/ in vitro did not affect the histamine-induced contraction in the absence of extracellular Ca(2+). Tolerance to the inhibitory action of ethanol developed selectively on contractile responses to histamine and phorbol 12,13-dibutyrate. Chronic ethanol administration produces tolerance to in vitro gallbladder contractility mediated by the Ca(2+) entry through L-type Ca(2+) channels linked with protein kinase C activation. PMID- 11901297 TI - Effect of SMP-500, a novel acyl-coA:cholesterol acyltransferase inhibitor, on the cholesterol esterification and its hypocholesterolemic properties. AB - We investigated the effects of SMP-500, a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, on ACAT activities in the liver and intestine, and in macrophages. We measured its effects on the serum cholesterol levels and hepatic cholesterol content in mice, rabbits and hamsters. SMP-500 inhibited ACAT activities in rabbit liver and small intestine microsomes with IC(50) values of 72 and 84 nmol/l, respectively, and acted as a competitive inhibitor of rabbit liver ACAT. SMP-500 potently inhibited cholesterol esterification in rat peritoneal macrophages (IC(50) = 15 nmol/l). In high-fat and high-cholesterol diet-fed mice and in high-cholesterol diet-fed rabbits, SMP-500 reduced the serum cholesterol levels and the hepatic cholesterol content. SMP-500 also reduced the serum and hepatic cholesterol in normal chow-fed hamsters in a dose-dependent manner. In all the animal models, SMP-500 reduced the hepatic free cholesterol content as well as the total and esterified cholesterol. Administered orally, SMP 500 had a direct inhibitory effect on hepatic ACAT activity. These results indicate that SMP-500 is a potent and competitive ACAT inhibitor and may have a therapeutic potential for treating hypercholesterolemia and atherosclerosis. PMID- 11901298 TI - Effect of 17beta-estradiol exposure on vasorelaxation induced by K(+) channel openers and Ca(2+) channel blockers. AB - 17beta-Estradiol has been shown to relax blood vessels partly through inhibition of Ca(2+) channels at supraphysiological concentrations; however, it is unknown whether acute exposure of the isolated artery rings to near physiological concentrations of sex steroid hormones could modulate the ionic channels that are involved in regulation of vascular tone. Brief incubation (20 min) with 17beta estradiol (1-3 nmol/l) did not alter the relaxant response to three blocking agents of L-type voltage-sensitive Ca(2+) channels, nifedipine, diltiazem and verapamil in either endothelium-intact or endothelium-denuded rat mesenteric artery rings. In contrast, 17beta-estradiol at 3 nmol/l significantly attenuated the relaxation induced by K(+) channel openers, cromakalim and pinacidil in endothelium-denuded rings. Similarly, preincubation with progesterone (3 nmol/l) inhibited pinacidil-induced relaxation with much less effect on cromakalim induced relaxation. It appears that 17beta-estradiol and progesterone attenuated the cromakalim- and pinacidil-induced relaxation in a different manner. These results suggest that acute exposure to female sex steroid hormones at near physiological levels may reduce the activity of ATP-sensitive K(+) channels in the rat arteries. PMID- 11901299 TI - A novel attribute of enoxaparin: inhibition of monocyte adhesion to endothelial cells by a mechanism involving cell adhesion molecules. AB - Enoxaparin is a low molecular weight heparin, widely accepted as anticoagulant or antithrombotic drug, and is likely to have a role in acute inflammation. To evaluate the anti-inflammatory potential of enoxaparin, we investigated the direct effect of the drug on the activation of endothelial cells. For this purpose we set up an in vitro system in which cultured valvular endothelial cells (VEC) activated by tumor necrosis factor alpha or lipopolysaccharide were exposed to a monocytic cell line; these conditions induced a significant adhesion of monocytes to VEC. Adhesion assays, ELISA, and flow cytometric analysis revealed that pretreatment with enoxaparin, at a relevant plasma concentration (16 microg/ml), acts upon activation of VEC by inhibition of lipopolysaccharide induced E-selectin expression and tumor necrosis factor stimulated ICAM-1 expression, thus reducing monocyte adhesion to VEC. These results suggest a novel function of enoxaparin, namely to protect VEC from activation and inhibiting the expression of cell adhesion molecules. PMID- 11901300 TI - Distribution and muscle-sparing effects of clenbuterol in hindlimb-suspended rats. AB - Based on their anabolic properties in skeletal muscles, beta-adrenergic agonists are of interest as potential countermeasures to microgravity-induced skeletal muscle atrophy. The levels of clenbuterol (Cb), a beta(2)-adrenergic agonist, in both plasma and skeletal muscle were higher in hindlimb-suspended rats than in their nonsuspended Cb-treated controls. Cb treatment was shown to help maintain the body weight in suspended rats, while reducing the amount of mesenteric fat. However, hindlimb suspension attenuated Cb's lipolytic effects. In skeletal muscle, the magnitude of response to unloading and Cb treatment followed a general regional pattern and was muscle and type specific. The highest magnitude of response to unloading was in predominantly slow-twitch muscles, and the least responsive were the predominately fast-twitch muscles. PMID- 11901301 TI - Desensitization of alpha(2)-adrenoceptors which regulate noradrenaline synthesis and release after chronic treatment with clorgyline in the rat brain. AB - The sensitivity of alpha(2)-adrenoceptors which regulate synthesis and release of noradrenaline was investigated in hippocampus, parietal cortex, and hypothalamus of rats treated with clorgyline. After administering a DOPA decarboxylase inhibitor, the in vivo tyrosine hydroxylase activity and the noradrenaline content were evaluated. Acute and chronic treatment with clorgyline led to both increases of noradrenaline levels and decreases of tyrosine hydroxylase activity, determined as the accumulation of DOPA. Whereas the alpha(2)-adrenoceptor agonist clonidine induced a similar reduction in tyrosine hydroxylase activity in the group subjected to the acute treatment and in the control group, it failed to do so after chronic clorgyline treatment. In hippocampal and cortical synaptosomes, a reduction in the sensitivity of alpha(2)-adrenoceptors which regulate [(3)H]noradrenaline release, reflected by the shift to the right of the concentration-effect curves for oxymetazoline, was also found after the repeated treatment. These results indicate a desensitization of alpha(2)-adrenoceptors after chronic treatment with clorgyline. PMID- 11901302 TI - STI571 (imatinib mesylate): the tale of a targeted therapy. AB - STI571 (imatinib mesylate) is an example of the successful development of a targeted agent. Its target is the constitutively active tyrosine kinase (p210bcr abl) in a hematologic neoplasm, chronic myelogenous leukemia (CML). The results in early clinical trials were remarkable and led to rapid approval by the Food and Drug Administration for clinical use of the STI571 in CML. This article reviews the pre-clinical and clinical development of this agent and also discusses some of the prevailing theories to explain the emerging problem of resistance. Future directions for this drug, possibly directed at other targets, are also discussed. PMID- 11901303 TI - Clostridium spores for tumor-specific drug delivery. AB - Insufficient blood supply of rapidly growing tumors leads to the presence of hypoxia, a well-known feature in solid tumors. Hypoxia is known to decrease the efficiency of currently used anti-cancer modalities like surgery, chemotherapy and radiotherapy. Therefore, hypoxia seems to be a major limitation in current anti-cancer therapy. The use of non-pathogenic clostridia to deliver toxic agents to the tumor cells takes advantage of this unique physiology. These strictly anaerobic, Gram-positive, spore-forming bacteria give, after systemic administration, a selective colonization of hypoxic/necrotic areas within the tumor. Moreover, they can be genetically modified to secrete therapeutic proteins like cytosine deaminase or tumor necrosis factor-alpha. The specificity of this protein delivery system can be further increased when expression is controlled by the use of a radio-inducible promoter, leading to increased spatial and temporal regulation of protein expression. This approach of bacterial vector systems to target protein expression to the tumor can be considered very safe since bacteria can be eliminated at any moment by the addition of proper antibiotics. The Clostridium-based delivery system thus presents an alternative therapeutic modality to deliver anti-tumor agents specifically to the tumor site. This high selectivity offers a major advantage in comparison with the classical gene therapy systems. PMID- 11901304 TI - COX-2 inhibitors in cancer treatment and prevention, a recent development. AB - Epidemiological and experimental studies have demonstrated the effect of non steroidal anti-inflammatory drugs (NSAIDs) in the prevention of human cancers. NSAIDs block endogenous prostaglandin synthesis through inhibition of cyclooxygenase (COX) enzymatic activity. COX-2, a key isoenzyme in conversion of arachidonic acid to prostaglandins, is inducible by various agents such as growth factors and tumor promoters, and is frequently overexpressed in various tumors. The contribution of COX-2 to carcinogenesis and the malignant phenotype of tumor cells has been thought to be related to its abilities to (i) increase production of prostaglandins, (ii) convert procarcinogens to carcinogens, (iii) inhibit apoptosis, (iv) promote angiogenesis, (v) modulate inflammation and immune function, and (vi) increase tumor cell invasiveness, although some studies indicated that NSAIDs have COX-2-independent effects. A number of clinical trials using COX-2 inhibitors are in progress, and the results from these studies will increase our understanding of COX-2 inhibition in both cancer treatment and prevention. The combination of COX-2 inhibitors with radiation or other anti cancer or cancer prevention drugs may reduce their side effects in future cancer prevention and treatment. Recent progress in the treatment and prevention of cancers of the colon, esophagus, lung, bladder, breast and prostate with NSAIDs, especially COX-2 inhibitors, is also discussed. PMID- 11901305 TI - Altered irinotecan metabolism in a patient receiving phenytoin. AB - The systemic exposure to the anticancer agent irinotecan (CPT-11) and its active metabolite SN-38 were 79 and 92% reduced, respectively, relative to literature data, by concomitant phenytoin therapy. This finding suggests that increased doses of CPT-11 should be given to patients treated simultaneously with these drugs, to achieve adequate levels of SN-38. PMID- 11901306 TI - Enhancement of fotemustine (Muphoran) cytotoxicity by amifostine in malignant melanoma cell lines. AB - Fotemustine (Muphoran, S10036), a nitrosourea derivative active in the treatment of malignant melanoma and primary brain tumors, was evaluated in combination with the free radicals cytoprotective agent amifostine (Ethyol, WR-2721) and its alkaline phosphatase (AP)-generated active metabolite WR-1065 in four human melanoma (RPMI-7950, SK-MEL2, SK-MEL5 and WM-115) and lung fibroblast (MRC-5) cell lines. No difference in AP activity was found among the melanoma cell lines, but AP was found to be significantly higher in MRC-5. For combination experiments, cell lines were first exposed to amifostine or WR-1065 for 15 min and then exposed to fotemustine for two cell doubling times. Non-cytotoxic amifostine and WR-1065 concentrations used (0.2 and 0.6 and 0.1 and 0.3 mmol/l, respectively) were deduced from clinically achieved plasma values. Interactions were analyzed from the variations in IC(50) of fotemustine induced by pre exposure of the cells to amifostine or WR-1065. In all melanoma cell lines, amifostine enhanced the cytotoxic activity of fotemustine as a significant decrease in IC(50) was observed. No significant difference was found between synergistic effects achieved with amifostine and WR-1065 given at half concentrations. No differential effect was found in the MRC-5 cell line as compared with the melanoma cell lines. Expression variation of O(6)-methylguanine methyltransferase was not found to be implicated in the interaction. The present results demonstrating that amifostine or its main active metabolite do not impair the cytotoxicity of fotemustine justify an extensive clinical evaluation of this combination in metastatic melanoma. PMID- 11901307 TI - Coupling of the antitumoral enzyme bovine seminal ribonuclease to polyethylene glycol chains increases its systemic efficacy in mice. AB - Bovine seminal ribonuclease (BS-RNase) is an antitumoral active enzyme exhibiting specific antitumoral action against a number of different cancer cell lines. However, its systemic use is limited by its pharmacokinetic properties and antigenicity. Therefore, it was conjugated to polyethylene glycol (PEG) chains to overcome these problems. Measurement of aspermatogenic effects of the preparation after s.c. injection and injection into the scrotum was chosen as a model for the distribution of the enzyme in the body mediated by the linkage to PEG chains. Additionally, the antigenicity of BS-RNase coupled to PEG chains (BS-RNase-PEG) was compared to that of free BS-RNase, as antigenicity is known to be one of the main obstacles in the use of protein-based drugs. BS-RNase-PEG caused aspermatogenic effects after systemic administration to mice in very low concentrations at which free BS-RNase is not effective. Moreover, BS-RNase possessed a very low antigenicity as long as it was coupled to the PEG chains. In order to investigate the antitumoral efficacy of BS-RNase-PEG in vivo, preliminary experiments on the effect of the conjugate on neuroblastoma growth in mice were performed in a UKF-NB-3 xeno-transplantate model, demonstrating a drastically increased anti-tumoral activity of the conjugate compared to the free enzyme. PMID- 11901308 TI - Eradication of osteosarcoma lung metastasis using intranasal gemcitabine. AB - We sought to determine whether gemcitabine, a new pyrimidine antimetabolite, could inhibit the growth of human osteosarcoma cells (OS) in vitro and in vivo. Four human OS cell lines (MG-63, TE-85, SAOS-2 and SAOS-LM7) were used to assess the activity of the drug in vitro. Gemcitabine caused growth inhibition and cell death in all four cell lines as measured using the MTT and colony-forming assays (IC(50) = 6.5 nM-9 microM and 7-14 nM, respectively). Using our newly developed human SAOS-LM7 OS lung metastasis mouse model, we assessed the in vivo activity of gemcitabine given i.p. and intranasally (i.n.). Mice were treated twice weekly for 3 weeks and then once weekly for 3 weeks using either i.p. or i.n. gemcitabine starting 4 weeks after tumor cell injection. The i.p. injection, at 120 mg/kg, resulted in a decrease in lung weights and the size of the nodules. However, no significant reduction in the number of metastatic nodules was seen (control median: >200 versus gemcitabine median: 150, p = 0.084). In contrast, the number of lung metastases was significantly decreased in mice that received i.n. gemcitabine at 15 (median: 1; range: 0-115, p<0.005) and 12 mg/kg (median: 41; range: 7-163, p = 0.005) when compared with control mice (median: >200). Intranasal therapy is a non-invasive method of drug delivery and has the advantage of targeting the lung, resulting in a higher drug concentration in the tumor area. In our study, i.n. instillation of gemcitabine inhibited the growth of lung metastases at an 8- to 10-fold lower dose than that used i.p. and appeared to be more effective in eradicating OS lung nodules. Because the lung is the most common site of OS metastasis, our data suggest that i.n. gemcitabine may be a novel therapeutic approach in the treatment of OS lung metastases. PMID- 11901309 TI - Pre-irradiation semi-intensive chemotherapy with carboplatin and cyclophosphamide in malignant glioma: a phase II study. AB - We undertook a phase II trial in 17 patients with malignant glioma and large measurable disease to assess response rate and survival with pre-irradiation chemotherapy, using higher doses than standard, trying to improve the outcome. Patients characteristics were: male/female 10/7, age 49 (range 23-59), median Karnofsky index 90% (range 70-100), glioblastoma multiforme/anaplastic astrocytoma 14/3. Treatment consisted of 2 cycles of carboplatin 200 mg/m(2) days 1-3 (or AUC x 8, total dose) plus cyclophosphamide 1000 mg/m(2) days 1-3. One partial response (6.5%) and two stabilizations (13.5%) were observed after pre irradiation chemotherapy. Twelve out of 15 patients (80%) progressed after chemotherapy. Median survival time was 7.6 months and the survival at 1 year was 33%. Main toxicity was hematologic in the first cycle: neutropenia grade 4 in 100%; thrombocytopenia grade 4 in 73% and grade 3 in 27%; anemia grade 3 in 7%; in the second cycle: neutropenia and thrombocytopenia grade 4 in 100% and anemia grade 3 in 50%). No toxic death was related to treatment. This regimen showed limited activity in malignant glioma with large residual disease after surgery or biopsy. PMID- 11901310 TI - Phase II trial of 9-nitrocamptothecin (RFS 2000) for patients with metastatic cutaneous or uveal melanoma. AB - The camptothecin derivative 9-nitrocamptothecin (9-NC) has demonstrated clinical activity in patients with ovarian and pancreatic carcinomas. Preclinical studies have shown promising activity of 9-NC for melanoma. We have thus conducted a phase II clinical trial of 9-NC for patients with metastatic cutaneous and uveal melanoma. Twenty-eight patients were enrolled in the trial, with diagnoses evenly divided between the two types of melanoma. 9-NC was administered orally at a starting dose of 1.5 mg/m(2)/day for 5 consecutive days of each week. No complete or partial responses were observed. Stabilization of disease was achieved in four individuals (15%) for durations of 3, 4, 6 and 8 months. Hematologic toxicity was moderate. Gastrointestinal side effects were common with 43% of the patients experiencing grade 3 or 4 diarrhea and 18% reporting grade 3 or 4 vomiting. In contrast to other 9-NC clinical trials, no patients developed chemical cystitis with gross hematuria. We conclude that, in keeping with the general chemoresistance of melanoma, 9-NC at the dose and schedule studied in this trial is significantly toxic and is not active for metastatic melanoma of cutaneous or uveal origin. PMID- 11901311 TI - Individualized long-term chemotherapy for recurrent ovarian cancer after failing high-dose treatment. AB - Chemotherapy for recurrent ovarian carcinoma (ROC) produces response rates of 10 80% depending on the prevalence of platinum resistance. Most patients relapse within 1 year and median progression-free survival is generally no more than 6 months. Newly developed ATP chemosensitivity assays (ATP-TCA) offer the opportunity for individualized therapy and have shown promising results compared to standard regimens. We report on an unusual case of long-term survival in a patient with stage III c ovarian cancer failing postoperative platinum-based high dose treatment who subsequently underwent repeated chemotherapy over a period of 4 years. The chemotherapy protocol was selected by pretherapeutic ex vivo ATP based chemosensitivity testing of autologous tumor tissue. To our knowledge, this is one of the few cases of ROC in which partial remissions using conventionally dosed chemotherapy were achieved repeatedly despite a unfavorable relapse-free interval after high-dose chemotherapy for primary disease. We conclude that ATP TCA-directed chemotherapy for ROC can select active and tolerable regimens even in difficult therapeutic situations in which no standards recommendation exists. PMID- 11901312 TI - The role of glucocorticoids in the treatment of fulminant hepatitis induced by dacarbazine. AB - Dacarbazine (DTIC) is commonly used for the treatment of malignant melanoma and Hodgkin's disease. A very rare complication of this cytotoxic agent is acute vascular hepatic damage, e.g. veno-occlusive disease or Budd-Chiari syndrome. The pathophysiological mechanism involved seems to be an immune reaction. This complication is frequently fatal. We report a patient who developed severe hepatic failure following DTIC treatment who responded favorably to treatment with glucocorticosteroid. PMID- 11901314 TI - Time to laparotomy for intra-abdominal bleeding from trauma does affect survival for delays up to 90 minutes. AB - OBJECTIVE: We examined the relationship between survival and time in the emergency department (ED) before laparotomy for hypotensive patients bleeding from abdominal injuries. METHODS: Patients in the Pennsylvania Trauma Systems Foundation trauma registry with isolated abdominal vascular, solid organ, or wall injuries grade 3 to 6 and hypotension were identified. Deaths were predicted from the prehospital time, systolic blood pressure (SBP) on ED admission, and time in the ED before either laparotomy or ED death. RESULTS: Two-hundred forty-three patients met the criteria. SBP ranged from 30 to 90 mm Hg. Time to the ED ranged from 7 to 185 minutes. Time in the ED ranged from 7 to 915 minutes. Overall, 98 patients died (40%). The risk ratio for the SBP increased, as expected, as SBP dropped. The risk ratio for time spent in the ED before laparotomy increased until 90 minutes, then significantly decreased below all earlier values. Logistic regression on the 165 patients spending 90 minutes or less in the ED showed that the probability of death increased with time in the ED. The increase was as much as 0.35% per minute. CONCLUSION: Among patients in a trauma registry who were hypotensive on arrival in the ED and had major injuries isolated to the abdomen requiring emergency laparotomy, the probability of death showed a relationship to both the extent of hypotension and the length of time in the ED for patients who were in the ED for 90 minutes or less. The probability of death increased approximately 1% for each 3 minutes in the ED. PMID- 11901313 TI - Hypoxia is not the sole cause of lactate production during shock. AB - BACKGROUND: Traditionally, elevated blood lactate after hemorrhage is interpreted as tissue hypoperfusion, hypoxia, and anaerobic glycolysis. The severity and duration of the increase in blood lactate correlate with death. Recent in vitro studies indicate that epinephrine stimulates lactate production in well oxygenated skeletal muscle by increasing activity of the Na+-K+-adenosine triphosphatase (ATPase), which derives a significant amount of adenosine triphosphate from glycolysis. Using in vivo microdialysis, we tested whether inhibiting the Na+-K+ pump with ouabain could reduce muscle lactate production during local exposure, via the microdialysis probe, to epinephrine or during hemorrhage in rats. METHODS: Microdialysis catheters were placed in the muscle of both thighs of pentobarbital-anesthetized male Sprague-Dawley rats (275-350 g) and perfused (1 microL/min) with Krebs-phosphate buffer (pH 7.4) containing ethanol (5 mmol/L) to permit assessment of changes in local blood flow. To inhibit the Na+-K+-ATPase, ouabain (2-3 mmol/L) was added to the perfusate of one leg. In one series of studies, epinephrine was added to the perfusate. In another series, rats were hemorrhaged to a mean arterial pressure of 45 mm Hg for 30 minutes, followed by resuscitation with shed blood and 0.9% sodium chloride. Dialysate fractions were analyzed for lactate and ethanol fluorometrically. RESULTS: Lactate rose during epinephrine exposure or during hemorrhage and resuscitation. Treatment with ouabain reduced dialysate lactate concentration significantly in both series of studies. Local blood flow was reduced by either epinephrine or hemorrhage, but returned toward baseline afterward. Ouabain had no apparent effect on local blood flow. CONCLUSION: Increased Na+-K+ATPase activity during epinephrine treatment or hemorrhage contributes to muscle lactate production. Hypoxia is not necessarily the sole cause of hyperlactatemia during and after hemorrhagic shock. PMID- 11901315 TI - Is delayed laparotomy for blunt abdominal trauma a valid quality improvement measure in the era of nonoperative management of abdominal injuries? AB - BACKGROUND: Review of hemodynamically stable patients who undergo laparotomy for trauma greater than 4 hours after admission is an American College of Surgeons quality improvement filter. We reviewed our recent experience with patients who underwent laparotomy for trauma greater than 4 hours after admission to evaluate the reasons for delay, and to determine whether they were because of failure of nonoperative management or other causes. METHODS: The registry at our Level I trauma center was searched from January 1998 through December 2000 for patients who required a laparotomy for trauma greater than 4 hours after admission. Of 3,369 admitted blunt trauma patients, 90 (2.7%) underwent laparotomy for trauma, of which 26 (29%) were identified as delayed laparotomies greater than 4 hours after admission. RESULTS: The most common mechanism of injury was motor vehicle crash, the mean Injury Severity Score was 18, and 65% of the patients had significant distracting injuries. Five patients had laparotomy greater than 24 hours after admission. The average time to the operating room in the remaining patients was 8.6 hours. Clinical examination (61%) findings were the most common indication for operation. Gastrointestinal (GI) tract injury was the most common injury associated with delay in laparotomy (58%). CONCLUSION: GI tract injuries are the predominant injury leading to delayed laparotomy for blunt trauma (58%). Failed nonoperative management of solid organ injuries occurred less frequently (15%). Future efforts should concentrate on earlier identification of GI tract injury. Delayed laparotomy for blunt abdominal trauma is a valid quality improvement measure. PMID- 11901316 TI - Slow channel calcium inhibition blocks proinflammatory gene signaling and reduces macrophage responsiveness. AB - BACKGROUND: This study investigates the possible intracellular mechanisms responsible for calcium antagonist protection in tissue-fixed macrophages, a central modulator of the proinflammatory phenotype. METHODS: Rabbit alveolar macrophages were exposed to lipopolysaccharide in the presence of different specific calcium antagonists. Cellular and nuclear protein were extracted and analyzed by Western blot for the phosphorylated forms of PYK2, ERK 1/2, and p38, and nuclear translocation of NF-kappaB and AP-1. Tumor necrosis factor-alpha (TNF alpha) expression was measured by an L929 bioassay on cellular supernatants. Statistical analysis was performed by unpaired Student's t tests. RESULTS: Cells pretreated with 100 to 500 micromol/L of diltiazem or 50 to 100 micromol/L of verapamil, both slow channel calcium blockers, led to dose-dependent reductions in lipopolysaccharide-induced PYK2 and ERK 1/2 phosphorylation, and nuclear translocation of AP-1 when compared with controls (p < 0.05). Neither inhibitor had any significant effect on p38 or NF-kappaB translocation. EGTA an extracellular calcium chelator, had no significant effect on any intracellular process studied. A dose-dependent reduction in TNF-alpha production was demonstrated with diltiazem and verapamil (p < 0.05), with no effect induced by EGTA. CONCLUSION: Slow channel calcium influx is essential for optimal intracellular signaling through PYK2 and ERK 1/2. This reduced intracellular signaling correlated with reduced AP-1 translocation and TNF-alpha production. Extracellular calcium chelation had no significant effect on intracellular signaling or TNF-alpha production. This study further elucidates the protective mechanism of action of calcium channel blockade by diltiazem and verapamil by reducing intracellular calcium release and down-regulating the excessive proinflammatory phenotype. PMID- 11901317 TI - Activated polymorphonuclear leukocytes enhance production of leukocyte microparticles with increased adhesion molecules in patients with sepsis. AB - BACKGROUND: Leukocyte microparticles (MPs) derived from polymorphonuclear leukocytes (PMNLs) have been recently found to be activators of vascular endothelium in vitro. The precise role of leukocyte MPs has not been clarified in patients suffering severe insult. The objective of this study was to evaluate production of leukocyte MPs and expression of adhesion molecules on the MP surface in patients with sepsis. METHODS: Twenty-one patients with severe infection (fulfilling the criteria of sepsis with serum C-reactive protein > 10 mg/dL) and 21 healthy volunteers were included as study subjects. Production of leukocyte MPs, expression of CD11b on the MPs, and oxidative activity of PMNLs were measured by flow cytometry in the presence and absence of formyl-methionyl leucyl-phenylalanine. CD11b expression was evaluated according to the MP size (more than, equal to, or less than 1.0 microm). Soluble E-selectin, thrombomodulin, and PMNL elastase were also measured in blood. RESULTS: Production of leukocyte MPs and superoxide production in PMNLs with and without formyl-methionyl-leucyl-phenylalanine increased significantly in patients with sepsis in comparison with production in normal volunteers. In patients with sepsis, expression of CD11b was also markedly enhanced on MPs less than 1.0 microm in diameter in comparison with expression in control subjects. Levels of soluble E-selectin, thrombomodulin, and PMNL elastase in blood were significantly increased in patients with sepsis. We succeeded in detecting leukocyte MPs visually by fluorescence microscopy. CONCLUSION: Activated PMNLs enhance production of leukocyte MPs with increased adhesion molecules in patients with sepsis. Activated leukocyte MPs may play a role in the pathogenesis of endothelial activation and leukocyte-endothelium interaction in the presence of sepsis. PMID- 11901318 TI - Free hemoglobin enhances tumor necrosis factor-alpha production in isolated human monocytes. AB - BACKGROUND: A systemic inflammatory response (SIR) is seen in approximately 75% of patients with complex blunt liver injuries treated nonoperatively. Many feel this response is caused by blood, bile, and necrotic tissue accumulation in the peritoneal cavity. Our current treatment for these patients is a delayed laparoscopic washout of the peritoneal cavity, resulting in a dramatic resolution of the SIR. Spectrophotometric analysis of the intraperitoneal fluid has confirmed the presence of high concentrations of free hemoglobin (Hb). We hypothesize that free Hb enhances the local peritoneal response by increasing tumor necrosis factor-alpha (TNF-alpha) production by monocytes, contributing to the local inflammatory response and SIR. METHODS: Monocytes from five healthy volunteers were isolated and cultured in RPMI-1640 for 24 hours. Treatment groups included saline controls, lipopolysaccharide ([LPS], 10 ng/mL, from Escherichia coli), human Hb (25 microg/mL), and Hb + LPS. Supernatants were analyzed by enzyme-linked immunosorbent assay. Student's t test with Mann-Whitney posttest was used for statistical analysis with p < or = 0.05 considered significant. RESULTS: Free Hb significantly increased TNF-alpha production 915 +/- 223 pg/mL versus saline (p = 0.02). LPS and Hb + LPS further increased TNF-alpha production (2294 pg/mL and 2501 pg/mL, respectively, p < 0.001) compared with saline controls. CONCLUSION: These data confirm that free Hb is a proinflammatory mediator resulting in the production of significant amounts of TNF-alpha. These in vitro findings support our clinical data in which timely removal of intraperitoneal free hemoglobin helps prevent its deleterious local and systemic inflammatory effects in patients with complex liver injuries managed nonoperatively. PMID- 11901319 TI - The role of surveillance duplex scanning in preventing venous thromboembolism in trauma patients. AB - BACKGROUND: This study was performed to determine the role of duplex scanning in preventing pulmonary embolism (PE), the correlation of venous thromboembolism (VTE) risk score with the incidence of deep venous thrombosis (DVT), and patients who may benefit from surveillance duplex scanning. METHODS: Age, sex, Injury Severity Score (ISS), VTE score, length of stay, diagnoses, and bleeding risk were recorded from the trauma registry in patients who had a duplex scan from 1995 to 2000. RESULTS: There were 1,513 duplex scans obtained (10,141 trauma admissions), 253 (2.5%) cases of DVT (52% above-knee, 8% upper extremity), and 30 cases of PE (0.3%). Only 5 of 21 duplex scans were positive in PE patients. DVT patients were older (52.9 vs. 46.7 years), with higher ISS (24.0 vs. 20.8) than patients without DVT. Regression analysis showed poor correlation between VTE score and DVT incidence (r2 = 0.27). Univariate analysis identified age, ISS, and VTE score as risk predictors for DVT. CONCLUSION: Adherence to an evidence-based VTE prophylaxis protocol is more important than surveillance duplex scanning in preventing VTE in trauma patients. PMID- 11901320 TI - Blood culturing practices in a trauma intensive care unit: does concurrent antibiotic use make a difference? AB - BACKGROUND: Febrile trauma patients have repeated blood cultures drawn during a prolonged hospitalization. We examined the diagnostic yield of blood cultures in severely injured patients to determine whether concurrent antimicrobial therapy or prophylactic administration of antibiotics affects blood culture growth. We also determined how rapidly growth changed to determine whether total numbers of blood cultures could be decreased. The hypotheses of the study were that concurrent antimicrobial administration affects blood culture yield, prophylactic administration alters the culture result, and repetitive culturing is unnecessary. METHODS: A retrospective chart review of trauma patients with minimum Injury Severity Score of 15 and minimum 5-day intensive care unit length of stay was performed. The dates and results of blood cultures and antibiotic type and administration dates were recorded. "Prophylactic" antibiotics were defined as antibiotics administered on admission to the unit. Computer software was used to match the blood culture date to the period of antimicrobial administration. Categorical data were compared using Fisher's exact test. RESULTS: Two hundred fifty-eight patients met entry criteria, and 208 charts were complete for review. One hundred twenty-nine patients had 347 sets of blood cultures drawn. The positive blood culture rate was 10.8% in patients off antibiotics, and 13.9% in patients on antibiotics (p = 0.68). All prophylactic antibiotics included a beta-lactam. Only 18% of positive blood cultures in patients receiving prophylactic antibiotics were sensitive to beta-lactams as opposed to 59% sensitivity in those who did not receive prophylaxis (p = 0.03). One hundred seventy-six sets of blood cultures were performed after an initial positive culture. Only three patients with an initial positive culture had a second positive culture with a different organism. The mean time to culturing a new organism after initial growth was 19 days. CONCLUSION: Concurrent antimicrobial administration does not alter blood culture yield. Prophylactic administration alters the type of organism cultured. Little new information is gained from repetitive culturing. PMID- 11901321 TI - The impact of a repealed motorcycle helmet law in Miami-Dade County. AB - OBJECTIVE: To study the impact of helmet nonuse in motorcycle crashes after the repeal of a mandatory helmet law in the state of Florida. METHODS: We prospectively studied all patients evaluated at the University of Miami/Jackson Memorial Medical Center from July 1, 2000, through December 31, 2000, involved in motorcycle crashes, and compared them with those seen during the same time period the year before the helmet law change. RESULTS: In 1999, before the repeal of the helmet law, there were 52 cases evaluated at our center compared with 94 after the law change. Helmet usage decreased from 1999 (83%) to 2000 (56%). The number of brain injuries (Abbreviated Injury Scale score > or = 2) during this same time period increased from 18 to 35, and the number of fatalities from 2 to 8. CONCLUSION: The repeal of a motorcycle helmet law significantly increased the number and severity of brain injuries admitted to our trauma center. PMID- 11901322 TI - Aortic intimal injuries from blunt trauma: resolution profile in nonoperative management. AB - OBJECTIVE: To provide preliminary data on the resolution profile of aortic intimal injuries treated nonoperatively and on the safety of nonoperative management of these injuries. METHODS: Five blunt trauma patients diagnosed by transesophageal echocardiography (TEE) with traumatic intimal injury of the aorta were assigned to nonoperative management. This included beta-blockade to maintain systolic blood pressure between 80 and 90 mm Hg and heart rate between 60 and 80 beats/min, serial TEE studies, and invasive monitoring in the intensive care unit. The evolution of injury, the effectiveness of nonoperative treatment, and the potential need for an operative intervention were monitored. RESULTS: The patients had a mean Injury Severity Score of 32 and sustained multiple associated thoracic and extrathoracic injuries. Aortic injuries were located at the level of the ligamentum arteriosum and in the descending aorta adjacent to the diaphragm in three and two patients, respectively. The mean size of injury was 12.5 mm (range, 5-20 mm) and a thrombus attached to the endothelium was present in three of the five patients. Complete resolution of injury occurred within 9.4 +/- 6.6 days (range, 3-19 days). All patients remained hemodynamically stable and adequately perfused. All demonstrated progressive resolution of their aortic intimal injuries. No complications related to the aortic injuries were identified during a mean follow-up of 16.8 months. CONCLUSION: This small series suggests that aortic intimal injuries smaller than 20 mm in hemodynamically stable patients treated with beta-blockade resolve within several days. This approach appears safe when monitored by serial TEE studies performed by experienced experts, and continuous invasive hemodynamic monitoring. PMID- 11901323 TI - Acute and long-term clinical significance of myocardial contusion following blunt thoracic trauma: results of a prospective study. AB - BACKGROUND: The clinical significance of myocardial contusion (MC) following blunt thoracic trauma is still unknown. Accordingly, in this prospective study in a regional trauma center we investigated the acute and long-term clinical significance of MC. METHODS: One-hundred eighteen patients with manifest or suspected blunt thoracic trauma were evaluated for cardiac injury. Initial assessment was based on results of electrocardiographic, echocardiographic, and serial enzyme measurements. A follow-up (FU) assessment conducted at 3 and 12 months, respectively, was performed using electrocardiography, echocardiography, and a bicycle ergometry exercise test. RESULTS: Of the 118 patients in the total sample, 67 (56.8%) required admission to the surgical intensive care unit (SICU) because of the severity of their noncardiac injuries. Fourteen patients of the total sample (11.8%) were diagnosed with MC and 13 of these (92.9%) belonged to the intensive care admissions, thus representing an incidence of 19.4% in this patient group. During the hospitalization period none of the patients with MC experienced acute cardiac complications. Eighty-six patients (72.0%) were assessed at FU. No new pathologies were found except in one patient. Exercise testing revealed no ECG abnormalities and none of the patients experienced limitations on the bicycle ergometer due to a cardiac cause. CONCLUSIONS: The incidence of 19.7% of MC in our patients with blunt chest injury requiring intensive care treatment indicates that this condition is frequent in polytraumatized patients. Outcome and prognosis in patients with MC is favorable and, thus, routine cardiac work-up is not indicated. Specific diagnostic and therapeutic measures should be limited to cases where cardiac complications develop. PMID- 11901325 TI - Evaluation of staff workload during resuscitation of trauma patients. AB - OBJECTIVE: Evaluating the medical staff workload during resuscitation of trauma patients is one of the important quality assurance activities to provide adequate medical manpower, especially for patients with life-threatening or severe injuries. Nevertheless, there is no method available to measure and calculate the amount of workload during resuscitation. We sought to develop a new framework of Workload Scoring System (WSS) to evaluate and quantify the medical staff workload during resuscitation. METHODS: From July 1996 to July 1998, the records of 11,800 trauma patients were prospectively collected from our computer-stored medical record system. The Workload Scoring System points with reference to age, different triage category on the basis of triage version of the Revised Trauma Score (RTS), level category on the basis of Injury Severity Score (ISS), and Abbreviated Injury Scale (AIS) in six body regions were calculated to survey the medical staff workload. RESULTS: The WSS points were 18.51 +/- 0.80 for triage I, 11.88 +/- 0.17 for triage II, and 6.90 +/- 0.04 for triage III trauma patients. The WSS points were 23.10 +/- 0.67 for Level I, 20.34 +/- 0.25 for Level II, 12.87 +/- 0.08 for Level III, and 6.03 +/- 0.02 for Level IV trauma patients. There were statistically significant differences among triage I, II, and III trauma patients, and among Level I, II, III, and IV trauma patients (p < 0.01). The worse the physiologic status and the greater the anatomic damage, the more medical staff workload was needed. Multiple regression with linear model may predict WSS points as an equation of -8.920 + 1.375 ISS + 1.785 RTS + 0.424 Age (r2 = 0.621), which accounts for 62.1% of the variance in WSS points. CONCLUSION: WSS provides a valuable tool to measure and quantify the medical staff workload during resuscitation as a function of -8.920 + 1.375 ISS + 1.785 RTS + 0.424 Age. The greatest benefit of this methodology is to forecast the expected medical staff workload to allocate sufficient medical manpower to provide the desired trauma care. PMID- 11901324 TI - Impact of discontinuing a hospital-based air ambulance service on trauma patient outcomes. AB - BACKGROUND: The clinical benefit of aeromedical transportation of injured patients in the civilian population has been debated. The purpose of this study was to examine the effects of discontinuing a hospital-based helicopter transport program on trauma patient outcomes, with the hypothesis that the loss of an air ambulance would result in increased transport time and increased mortality among severely injured patients. METHODS: Data on injury severity and patient outcomes were collected prospectively for the 12 months immediately preceding and 24 months following discontinuation of the helicopter ambulance service. Transport time, mortality rate, and hospital length of stay was compared. RESULTS: The number of trauma patient admissions decreased 12%, with a 17% decrease in admissions of severely injured patients. Transport time decreased, with no change in mortality. CONCLUSION: Discontinuation of a hospital-based air ambulance service did not increase transport time or increase mortality for trauma patients. PMID- 11901326 TI - Functional results of unilateral mandibular condylar process fractures after open and closed treatment. AB - BACKGROUND: This retrospective study compared the functional results of unilateral mandibular condylar process fractures treated either by open reduction or by closed treatment. METHODS: Sixty-six patients with unilateral mandibular condylar process fractures were reviewed. Thirty-six patients received open reduction, and the other 30 underwent closed treatment (intermaxillary fixation only). Each group was further divided into condylar and subcondylar subgroups according to fracture level. The functional outcome was evaluated by posttreatment occlusion status, maximal mouth opening, facial symmetry, chin deviation, and temporomandibular joint symptoms. RESULTS: Patients undergoing closed treatment exhibited more condylar motility than those treated by open reduction. Patients in the condylar subgroup with open reduction presented less chin deviation (21.43%) compared with those with closed treatment (56.25%; p = 0.072). Although a greater severity of subcondylar fractures existed in patients treated with open reduction, patients treated with open reduction or closed treatment did not reveal a significantly functional difference. CONCLUSION: The present study revealed that patients with condylar neck or head fractures gained more benefits from open reduction in terms of chin deviation and temporomandibular joint pain. For subcondylar fractures, open reduction provides satisfactory functional results in patients with severely displaced fractures. PMID- 11901327 TI - Complications of rigid intramedullary rodding of femoral shaft fractures in children. AB - BACKGROUND: Intramedullary rodding of femoral shaft fractures has been frequently performed in adults, but until recently rarely in children. It was the purpose of this study to investigate the experience with this treatment method at a pediatric trauma center. METHODS: From 1987 to 1998, 54 children were treated for traumatic femoral fractures with intramedullary rods at a major pediatric trauma center. The average age was 15 years 3 months, ranging between 11 years 4 months and 17 years 11 months. The average follow-up was 5 years 3 months, ranging from 20 months to 10 years 1 month. RESULTS: All of the fractures occurred secondary to trauma and the most common anatomic fracture site was the femoral midshaft. Complications encountered included 8 instances of minor limb length discrepancy, 11 instances of discomfort because of rod prominence, 1 case of avascular necrosis of the femoral head, 2 instances of heterotopic ossification over the rod tip, 1 broken rod, and 3 cases that demonstrated decreased external rotation of the affected limb. One child developed osteomyelitis after intramedullary rodding for a fracture previously treated with external fixation. There were no cases of surgically induced nonunion or malunion and only one delayed union secondary to infection. CONCLUSION: Results of this series demonstrate intramedullary rodding to be an effective treatment modality for femoral fractures in skeletally mature children. In children with open femoral physes, rigid rodding should be avoided because of the small but serious occurrence of avascular necrosis of the femoral head. Intramedullary rodding is not recommended in children initially treated with external fixation because of the increased risk of infection. PMID- 11901328 TI - Reduction of severe wrist injuries in snowboarding by an optimized wrist protection device: a prospective randomized trial. AB - BACKGROUND: The benefits of sport are well recognized, but many activities carry a sport-specific injury risk. Snowboarding has become an increasingly popular winter sport in Austria in recent years, with an estimated 900,000 participants annually. Roughly 6,000 of these suffer from injury and up to 2,000 sustain moderate or severe wrist injuries (mainly fractures of the distal radius and epiphysiolyses). METHODS: We conducted a prospective, randomized, controlled trial to test the protective effect of a wrist protector, which differs in position, stiffness, length, and fixation from conventional protectors. Seven hundred twenty-one snowboarders were randomized into two groups. The risk factors and the injuries that occurred were registered by questionnaires and, in case of medical treatment, by medical reports. Time until injury (in half-days) was compared by the proportional hazards model. RESULTS: Nine severe wrist injuries were sustained in the unprotected control group and only one in the protected group (hazard ratio, 0.13; 95% confidence limits, 0.02, 1.04). Twelve snowboarders of the protector group secretly discarded their protectors during the trial (including the snowboarder who suffered the one and only severe wrist injury of this group). A per-protocol analysis was therefore performed, which demonstrated a more accentuated result (p = 0.003). There was no statistically significant increase in the incidence of other types of injury. Experience was shown to be a further protective factor. CONCLUSION: We recommend the use of a wrist protector, particularly for novices participating in this sport. As in other domains of medicine, preventive measures can decrease morbidity also in terms of sport injuries. PMID- 11901329 TI - Late whiplash syndrome: correlation of brain SPECT with neuropsychological tests and P300 event-related potential. AB - BACKGROUND: The acceleration forces infringing the cervical spine in whiplash injury are frequently associated with multiple cerebral symptoms. The purpose of this study was to determine whether there is a correlation between cerebral perfusion findings, P300 recording (an electrophysiologic marker of cognitive ability), and neuropsychological tests in patients with whiplash injury. METHODS: Twenty patients with chronic whiplash injury underwent extensive clinical evaluation and neuropsychological testing. A brain single-photon emission computed tomography (SPECT) study using 99mTc-HMPAO was performed in all patients within 24 hours of neuropsychological evaluation. P300 event-related potentials were performed in 15 patients and in 9 normal volunteers. RESULTS: Thirteen of 20 patients had brain perfusion abnormalities on the SPECT studies, in one or more regions. Eight of 15 patients had abnormal P300 studies. Seven of eight patients with abnormal P300 had also an abnormal SPECT study. Seven of 15 patients had normal P300 results, 6 of them with a normal SPECT and 1 with SPECT abnormalities. There was no significant correlation between the SPECT findings or the P300 results and the scores of attention and working memory. There was, however, close agreement between the SPECT and P300. CONCLUSION: SPECT perfusion abnormalities in patients with chronic whiplash syndrome correlate well with P300 recording. The combination of these studies with neurocognitive and neurobehavioral tests may be useful in identifying a subgroup of patients having organic brain lesions. PMID- 11901330 TI - Using the SF-36 for characterizing outcome after multiple trauma involving head injury. AB - BACKGROUND: The purpose of this study was to evaluate the validity of the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) for examining outcomes after multiple trauma and to investigate whether the addition of items selected to measure cognitive function could improve the sensitivity of the SF-36 for identifying differences in outcomes for patients with and without head injury. METHODS: One thousand two hundred thirty patients discharged from 12 trauma centers were interviewed 1 year after injury. The interview included the SF-36 supplemented with four items chosen to assess cognitive function. RESULTS: The resulting cognitive function scale is internally consistent and measures a component of health that is independent of the dimensions incorporated in the SF 36. It correlates well with established measures of brain injury severity and discriminates among patients with and without brain injury. CONCLUSION: This study underscores the need to supplement the SF-36 with a measure of cognitive function when evaluating outcome from multiple trauma involving head injury. Further studies are needed to validate the specific items chosen for measuring cognitive function. PMID- 11901332 TI - Barrier precautions in trauma: is knowledge enough? AB - OBJECTIVES: The risk of blood and body fluid exposure and, therefore, risk of blood-borne disease transmission is increased during trauma resuscitations. Use of barrier precautions (BPs) to protect health care workers (HCWs) from exposure and infection has been codified in hospital rules and in national trauma education policy. Despite these requirements, reported rates of BP compliance vary widely. The reasons for noncompliance are not known. This study assesses self-reported rates of BP usage during resuscitations among trauma professionals, explores reasons for noncompliance, and compares self-reported compliance rates with actual observed compliance rates. METHODS: A survey regarding BPs was distributed to all HCWs involved in trauma resuscitations at our Level I trauma center. All surgical and emergency medicine residents as well as attending faculty from both disciplines and nursing staff were included in this study. A total of 161 surveys were distributed and 123 were returned. RESULTS: Most HCWs (114 of 123 [93%]) reported at least one exposure (usually intact skin contact) to blood or other body fluids. A considerable variation in the type of BP used was reported for those HCWs who reported use of BPs "all of the time." Of the HCWs who reported universal use of BPs, reported usage rates were as follows: gloves, 105 of 123 (85%); eyewear (no side protectors), 58 of 123 (47%); eyewear (side protectors), 20 of 123 (16%); gowns, 22 of 123 (18%); and masks, 5 of 123 (4%). The two most common reasons for noncompliance were "time factors" (61%) and "BPs are too cumbersome" (29%). Observed compliance rates were statistically significantly lower than self-reported rates in all BPs except gloves (p < 0.02). CONCLUSION: The wide variation in BP use and the gap between perceived and actual usage that we have observed suggest that the effectiveness of current educational approaches to ensure BP use is inadequate. PMID- 11901331 TI - Complications of plate fixation in metacarpal fractures. AB - BACKGROUND: The objective of this study is to assess the complications after open reduction and plate fixation of extra-articular metacarpal fractures. METHODS: We retrospectively reviewed the clinical and radiologic records of 129 consecutive patients with 157 metacarpal fractures treated by open reduction and internal fixation with plates between 1993 and 1999. Intra-articular fractures and fractures of the thumb metacarpal were excluded. Eighty-one patients (64 men and 17 women) with 104 fractures were available for review, at an average follow-up of 13.6 months (range, 6-27 months). RESULTS: Twenty-eight patients (35%) and 33 fractures (32%) had one or more complications, including difficulty with fracture healing (12 patients [15%]), stiffness (eight patients [10%]), plate loosening or breakage (seven patients [8%]), complex regional pain syndrome (two patients), and one patient who developed a deep infection. CONCLUSION: Despite technical advances in implant material, design, and instrumentation, plate fixation of metacarpal fractures remains fraught with complications and unsatisfactory results. PMID- 11901333 TI - The effect of posterior sag on the fixation stability of intertrochanteric hip fractures. AB - BACKGROUND: This study evaluates the effects of posterior sag on the fixation stability of intertrochanteric hip fractures. METHODS: A simulated, two-part intertrochanteric fracture was created in human cadaveric femurs. One of each pair was stabilized using a sliding hip screw in anatomic reduction and the other in 30 degrees of posterior sag. Measurements for load versus inferior head displacement, gapping, and shearing were made in axial and torsional loading. RESULTS: Initial axial and torsional loading showed no significant differences between the two groups. During cyclic loading, the osteotomy gap in the posteriorly angulated specimens decreased by 0.11 cm at 10 cycles (p = 0.006) and by 0.22 cm at 10,000 cycles (p = 0.33), corresponding to a 2-degree and 6-degree reduction in sag angle. Axial stiffness differed between the two groups: 10.3 N/mm for anatomic versus 6.7 N/mm (p = 0.002) for posteriorly angulated specimens. Loading to failure demonstrated no significant differences between the two groups. CONCLUSION: This study demonstrated that 30 degrees of posterior sag does not result in a significant difference in construct strength or stability. PMID- 11901334 TI - Incidence, severity, and patterns of intrathoracic and intra-abdominal injuries in motorcycle crashes. AB - BACKGROUND: Although severe head injuries have been reduced with helmet use, little has been done to address the severity of trauma to organs of the trunk in motorcycle crashes. We detail the frequency, severity, and pattern of intrathoracic and intra-abdominal injuries that may be helpful in the recognition and medical treatment of such injuries. METHODS: Diagnostic and treatment information from emergency room, hospital, and coroner records from a cohort of motorcyclists injured from 1991 to 1992 were obtained from 28 hospitals and 11 coroners in California. RESULTS: Multiple intrathoracic and intra-abdominal injuries were common, and the number and bilaterality of rib fractures were strongly associated with chances of injuries to the thoracic and abdominal organs. CONCLUSION: Patients with severe injury in one anatomic region of the trunk are very likely to have severe injuries in the same or other anatomic regions. These patients are best treated in trauma centers, where rapid diagnosis and treatment are possible. PMID- 11901335 TI - Superior mesenteric artery-duodenal fistula presenting as a late complication of an abdominal gunshot wound. PMID- 11901337 TI - Bilateral diaphragm rupture: a unique presentation. PMID- 11901336 TI - Delayed hemorrhage after blunt hepatic trauma: case report. PMID- 11901338 TI - Long-term results after laparoscopic repair of traumatic diaphragmatic hernias. PMID- 11901339 TI - Traumatic rupture of aorta should be ruled out in severe injuries from paragliding: report of three cases. PMID- 11901340 TI - Traumatic rupture of the innominate and left common carotid artery: case report. PMID- 11901341 TI - Delayed cardiac tamponade after blunt chest trauma in a child. PMID- 11901342 TI - Penetrating injury from a less-lethal bean bag gun. PMID- 11901343 TI - Lightning-induced injury on an airplane: coronal discharge and ball lightning. PMID- 11901344 TI - Magnetic resonance imaging of simultaneous bilateral quadriceps tendon rupture in a weightlifter: case report. PMID- 11901345 TI - Cervical epidural hematoma after spinal manipulation therapy: case report. PMID- 11901346 TI - Airbag-mediated craniocervical injury in a child restrained with safety device. PMID- 11901347 TI - A new screw catheter kit for the bedside treatment of chronic subdural hematomas. PMID- 11901348 TI - The use of chemical incapacitant sprays: a review. PMID- 11901349 TI - Arterial blood gas analysis in the initial evaluation of the nonintubated adult blunt trauma patient. PMID- 11901351 TI - Ultrasound probably has a bright future in the diagnosis of pneumothorax. PMID- 11901352 TI - Late thoracic outlet syndrome secondary to malunion of the fractured clavicle: case report and review of the literature. PMID- 11901353 TI - The image of trauma: dive tank explosion amputation. PMID- 11901354 TI - The Fawn-Hooded (FH/Wjd) rat: a genetic animal model of comorbid depression and alcoholism. AB - The Fawn-Hooded (FH/Wjd) rat is an inbred strain of rat that has been reported to exhibit both high immobility in the forced swim test and high voluntary ethanol intake, measures that have been periodically linked with depression and alcoholism in humans. The present paper will first present a survey of the literature and previously unpublished findings that bear on the question of whether FH/Wjd rats should be considered genetic animal models of depression and alcoholism. Subsequently, behavioral studies of the FH/Wjd rats, the non-drinking ACI/N strain, and their F1 and F2 intercrosses will be described. Under free choice conditions, the FH/Wjd rat drinks up to 6 g/kg 10% ethanol per day. This intake was sufficient to render the rats tolerant to the hypothermic effects of injected ethanol (2.5 g/kg). Rats that had been voluntarily drinking for at least 6 weeks also exhibited withdrawal-induced anxiety in the social interaction, elevated plus maze, and ultrasonic vocalization tasks. The FH/Wjd rat exhibits a 25-30% increase in alcohol intake when the alcohol is returned after a 24-h period of deprivation. It responds to drugs that are effective in humans with a reduction in alcohol intake. Therefore, the FH/Wjd rat meets most of the criteria for an animal model of alcoholism. Chronic antidepressant treatments correct several of the abnormalities exhibited by the FH/Wjd rats, including the exaggerated immobility in the forced swim test. Therefore, the FH/Wjd rats also fulfill some of the criteria for an animal model of depression. On the contrary, inbred ACI/N rats do not drink much alcohol voluntarily and are quite active in the forced swim test. The FH/Wjd and ACI/N rats were intercrossed to obtain the F1 and F2 progenies, which were then tested for alcohol intake and immobility. Alcohol intake and immobility were distributed in different patterns in the F1 and F2 progenies. Alcohol intake was intermediate in the F1 progeny, while immobility was closer to the FH/Wjd parents. In the F2 progeny, chi-square analyses indicated that the distributions were significantly different. In addition, there were no significant litter effects, indicating that maternal effects did not appear to occur. There were also no significant differences among rats with different coat colors, suggesting that the Fawn-Hooded phenotype can be separated from the measures of alcohol intake and immobility. We conclude that the FH/Wjd rat is a genetic animal model of depression and alcoholism, but that the two measures reflective of these states are under separate genetic controls. PMID- 11901355 TI - Analysis of polymorphisms in the olfactory G-protein Golf in major depression. AB - It is well established that G-proteins represent essential regulatory components in transmembrane signaling. The alpha subunit of the olfactory G-protein Golf (GNAL) maps to a region on chromosome 18 where linkage to affective disorders has been reported, as well as a parent-of-origin effect in affective disorders with some markers near the locus for the alpha subunit of the Golf gene. We investigated whether two polymorphisms in the alpha subunit of the Golf gene (A- >G in intron 3, and T-->G in intron 10) are associated with major depression in 176 major depressive patients compared with 145 healthy control subjects, and additionally tested for a parent-of-origin effect in separated gender groups. In the control group, we found a significant increase in the G-allele frequency of the intron 3 polymorphism in females (P=0.0036, odds ratio=2.13, 95% confidence interval=1.29-3.54, Fisher's Exact Test). In patients, we found a similar tendency for higher G-allele frequencies in females. Concerning the intron 10 polymorphism, no differences in the genotype or allele frequencies were detectable for any of the separated gender groups. Also, the total patient and control groups showed no differences in allele or genotype frequencies for any of the investigated polymorphisms. The results of this study agree with the reported parent-of-origin effects on chromosome 18, but do not support the hypothesis that the Golf gene is a major susceptibility factor for major depression. PMID- 11901356 TI - Manic-depressive illness: an association study with the inositol polyphosphate 1 phosphatase and serotonin transporter genes. AB - Association studies with candidate genes may contribute towards the understanding of the etiopathogenesis of bipolar disorder. Candidate genes in bipolar disorders are those related to aminergic neurotransmission, which is the target of the effects of antipsychotics and antidepressants, as well as genes related to signal transduction pathways, reporting the target for the mood-stabilizing effects of lithium. Association with such candidate genes may provide clues towards the understanding of the biological components of bipolar disorder. An association study was performed between the 5' regulatory region of the serotonin transporter gene (5-HTTLPR), the inositol polyphosphate 1-phosphatase gene (INPP1) and bipolar disorder using our sample of proband/parent trios. A total of 101 bipolar probands were considered eligible for the study. Since both parents had to be available, mean age at onset of bipolar disorder in probands was relatively young. However, the mean duration of illness and the number of episodes were consistent with a stable diagnosis. In our trios sample, the transmission disequilibrium test revealed no preferential transmission of alleles of the 5 HTTLPR and INPP1 from heterozygous parents to probands. Therefore, additional family-based data are warranted, possibly with a more complete subdivision of 5 HTTLPR alleles, since short and long alleles have recently been divided into four and six kinds of allelic variant, respectively, with significant ethnic differences in allele and genotype distributions. PMID- 11901357 TI - No association for D2 and D4 dopamine receptor polymorphisms and methamphetamine abuse in Chinese males. AB - The D2 and D4 dopamine receptors (DRD2 and DRD4) play major roles in the central effects of psychostimulants and in the reward system. Previous studies, although not all, have demonstrated associations between the DRD2 TaqI and the DRD4 exon III variable number tandem repeat (VNTR) polymorphisms and substance dependence. For this study, we have investigated the associations between these two polymorphisms and methamphetamine (MAP) dependence, as manifested in a Chinese male sample population. No significant difference was demonstrated for genotype or allele frequency when comparing MAP-dependent and control cases for the DRD2 TaqI and the DRD4 gene exon III VNTR polymorphisms, suggesting that these two polymorphisms do not play major roles in MAP dependence for our sample of Chinese males. PMID- 11901359 TI - Genetic analysis of the (CTG)n NOTCH4 polymorphism in 65 multiplex bipolar pedigrees. AB - A strong genetic association between the NOTCH4 locus on chromosome 6 and schizophrenia was recently reported. Based on the data suggesting overlapping susceptibility for schizophrenia and bipolar disorder, we genotyped the polymorphic (CTG)n encoding polyleucine repeat in exon 1 of NOTCH4 in 65 pedigrees ascertained for a genetic linkage study of bipolar disorder. In addition, we analyzed a subset of our pedigrees with psychotic features at this locus. We failed to find any association between the (CTG)n NOTCH4 polymorphism and either the bipolar or the psychotic bipolar phenotype in our 65 pedigrees. PMID- 11901358 TI - Fine mapping of the chromosome 2p12-16 dyslexia susceptibility locus: quantitative association analysis and positional candidate genes SEMA4F and OTX1. AB - A locus on chromosome 2p12-16 has been implicated in dyslexia susceptibility by two independent linkage studies, including our own study of 119 nuclear twin based families, each with at least one reading-disabled child. Nonetheless, no variant of any gene has been reported to show association with dyslexia, and no consistent clinical evidence exists to identify candidate genes with any strong a priori logic. We used 21 microsatellite markers spanning 2p12-16 to refine our 1 LOD unit linkage support interval to 12cM between D2S337 and D2S286. Then, in quantitative association analysis, two microsatellites yielded P values<0.05 across a range of reading-related measures (D2S2378 and D2S2114). The exon/intron borders of two positional candidate genes within the region were characterized, and the exons were screened for polymorphisms. The genes were Semaphorin4F (SEMA4F), which encodes a protein involved in axonal growth cone guidance, and OTX1, encoding a homeodomain transcription factor involved in forebrain development. Two non-synonymous single nucleotide polymorphisms were found in SEMA4F, each with a heterozygosity of 0.03. One intronic single nucleotide polymorphism between exons 12 and 13 of SEMA4F was tested for quantitative association, but no significant association was found. Only one single nucleotide polymorphism was found in OTX1, which was exonic but silent. Our data therefore suggest that linkage with reading disability at 2p12-16 is not caused by coding variants of SEMA4F or OTX1. Our study outlines the approach necessary for the identification of genetic variants causing dyslexia susceptibility in an epidemiological population of dyslexics. PMID- 11901361 TI - CYP2D6 polymorphisms and atypical antipsychotic weight gain. AB - Reports have linked atypical antipsychotics (AAPs) with weight gain. The polymorphic CYP2D6 involved in metabolism has been associated with medication morbidity. Eleven subjects receiving olanzapine were genotyped for CYP2D6 to examine the relationship between 2D6 and AAP weight gain. Using a linear regression, the dependent variable was percent change in body mass index (BMI). Genotype, dose and duration of treatment were independent. Genotype was significant (P<0.0097) for those with a *1/*3 or *4 genotype experiencing a larger percent BMI change than those with a *1/*1 genotype. This may be due to increased olanzapine concentrations leading to increased exposure, which may trigger AAP weight gain. PMID- 11901360 TI - Alpha2-macroglobulin exon 24 (Val-1000-Ile) polymorphism is not associated with late-onset sporadic Alzheimer's dementia in the Hungarian population. AB - Several lines of biochemical evidence support a role of alpha2-macroglobulin (A2M) in the pathogenesis of Alzheimer's dementia (AD). A2M participates in the general defence mechanism against proteinases and it is supposed to be involved in the degradation of beta-amyloid peptide (betaAP). Furthermore, A2M has been shown to reduce betaAP fibril formation, and it is upregulated in the acute-phase inflammatory response like the process occurring in the AD brain. The exon 18 splice acceptor deletion polymorphism and the exon 24 (Val-1000-Ile) GG genotype were reported to be associated with AD, but the results are contradictory. Since the Hungarian population is genetically distinct from the other European ethnic groups, we examined whether the risk for developing AD is increased in the A2M GG carriers. The interaction of apolipoprotein E (apoE) and A2M polymorphisms was also examined. The distribution of A2M genotypes and alleles in the entire data set was consistent with the previous negative observations in which A and G allelic frequencies were comparable in both groups (72% and 28% in the AD population, and 72% and 28% in the control population, respectively). The GG genotype was over-represented (14%) only in the apoE epsilon4 non-carrier subgroup of AD probands (7% in the control group), but the difference was not significant. Our data suggest that, although A2M has an important role in the AD specific neurodegenerative process, its exon 24 Val-1000-Ile polymorphism is not likely to be associated with late-onset sporadic AD in the Hungarian population. PMID- 11901363 TI - Repetitive-strain injury cases. PMID- 11901364 TI - So you want to be the boss! PMID- 11901365 TI - Complementary therapies for asthma. PMID- 11901366 TI - Disclosing personal information for research purposes. PMID- 11901367 TI - Pediatric case management for respiratory syncytial virus outcomes. PMID- 11901369 TI - Telephonic response to workers in crisis. PMID- 11901370 TI - Issues and considerations in pharmacology: a call for case managers. PMID- 11901371 TI - Tragedy teaches valuable lessons in the wake of 9-11. PMID- 11901372 TI - Creating partnerships for optimal transplant care. PMID- 11901373 TI - The science of collaboration. PMID- 11901374 TI - Controlling costs and improving diabetes care. AB - A simplification of the threat that diabetes poses to industrialized society is that the costs of inadequate treatment in terms of dollars, suffering, and functional impairment, despite increasing knowledge, are ever increasing. This increased threat is a result of the combined effects of the rising incidence of diabetes, the increasing costs of adequate care, and the perverse effects of a medical care system that fails to encourage lifelong patient commitment and behavior change. PMID- 11901375 TI - New interventions in diabetes with medical nutrition therapy. AB - Nutrition therapy has been the focus of diabetes management since before insulin was discovered.(1) Many theories and approaches have been recommended and reemerged over the years. Since the Diabetes Control and Complications Trial (DCCT) results were released in 1993, nutrition is considered the most critical and pivotal component of diabetes care in achieving blood glucose goals. We have seen increased emphases on individualized nutrition therapy and the dietitian as a true partner in diabetes care, research, and management.(1) Advances in nutrition therapy now center on methods to improve behavioral change because it is the major challenge facing people with diabetes. Access to nutrition therapy and self-management training is critical to improve clinical outcomes and reduce health care costs otherwise spent on clinic visits, expensive medications, emergency room visits, and hospitalizations.(1) PMID- 11901377 TI - Generational differences. PMID- 11901376 TI - New advances in the diagnosis and management of nausea and vomiting. AB - This article has three aims, the first of which is to provide an overview of the assessment and management of chronic nausea and vomiting. Second, it details the definitions, approach to assessment, diagnostic classifications, and treatment options of patients with nausea and vomiting. Finally, specific emphasis is placed on chronic drug refractory nausea and vomiting (gastroparesis) and the clinical cost and quality of life data associated with gastric electric stimulation. PMID- 11901378 TI - Early results of the Universal total wrist arthroplasty in rheumatoid arthritis. AB - Early results of 2 surgeons involved in a prospective study of the Universal total wrist prosthesis (KMI, San Diego, CA) are reported. Twenty-two prostheses were implanted in 19 patients for the treatment of severe rheumatoid arthritis. Two-year follow-up results of 8 wrists and 1-year follow-up results of 14 wrists were reviewed. Total arcs of motion (flexion-extension, radial-ulnar deviation, and pronation-supination) all improved significantly after arthroplasty. Individual motions that were most limited before surgery (extension, radial deviation, and supination) improved the most. Disabilities of the Arm, Shoulder, and Hand outcome scores improved 14 points at 1 year and 24 points at 2 years. Three prostheses (14%) were unstable and required further treatment; all 3 were in patients with highly active disease and severe wrist laxity. The Universal prosthesis provides a good early outcome in rheumatoid patients without severe preoperative wrist laxity. PMID- 11901379 TI - Volar fixation for dorsally displaced fractures of the distal radius: a preliminary report. AB - Using a volar approach to avoid the soft tissue problems associated with dorsal plating, we treated a consecutive series of 29 patients with 31 dorsally displaced, unstable distal radial fractures with a new fixed-angle internal fixation device. At a minimal follow-up time of 12 months the fractures had healed with highly satisfactory radiographic and functional results. The final volar tilt averaged 5 degrees; radial inclination, 21 degrees; radial shortening, 1 mm; and articular incongruity, 0 mm. Wrist motion at final follow-up examination averaged 59 degrees extension, 57 degrees flexion, 27 degrees ulnar deviation, 17 degrees radial deviation, 80 degrees pronation, and 78 degrees supination. Grip strength was 79% of the contralateral side. The overall outcome according to the Gartland and Werley scales showed 19 excellent and 12 good results. Our experience indicates that most dorsally displaced distal radius fractures can be anatomically reduced and fixed through a volar approach. The combination of stable internal fixation with the preservation of the dorsal soft tissues resulted in rapid fracture healing, reduced need for bone grafting, and low incidence of tendon problems in our study. PMID- 11901380 TI - Osteotomy for malunited fractures of the distal radius: a comparison of structural and nonstructural autogenous bone grafts. AB - Two cohorts of 10 patients who had a corrective osteotomy for a malunited fracture of the distal radius with a pi-shaped plate and screw fixation were compared retrospectively to see whether the outcome was affected by using a nonstructural cancellous bone graft compared with a trapezoidal corticocancellous bone graft. The indications for the osteotomy, surgical techniques, and postoperative rehabilitation were consistent and all surgical procedures were done by the same surgeon. All osteotomies healed without loss of the surgical correction. Follow-up radiographic and functional results were comparable between groups. Use of a nonstructural, cancellous only bone graft-appealing in its relative simplicity-seems safe and efficacious. PMID- 11901381 TI - The effects of dorsally angulated distal radius fractures on carpal kinematics. AB - A cadaver model was used in a biomechanical study of dorsally angulated distal radius fractures to evaluate alterations in carpal kinematics. Distal radius fractures were simulated by dorsal closing-wedge osteotomy and fixed with a custom-designed external fixator. A magnetic tracking device measured the carpal bone motions in several positions of dorsal angulation from neutral tilt to 30 degrees dorsal tilt. Changes in carpal alignment showed different patterns between each specimen consisting of a spectrum from dorsal subluxation of the entire carpus to adaptive dorsal carpal instability (DISI deformity). Components of carpal bone motion were altered markedly at all positions of dorsal angulation of the distal radius. The severity of the DISI deformity and related carpal instability correlated well with the alterations of carpal kinematics during wrist flexion and extension, whereas dorsal subluxation alone had a poor relationship with changes in carpal kinematics. The amount of DISI deformity and the degree of dorsal angulation of the radius may be prognostic factors when considering whether to perform a corrective osteotomy of the distal radius. PMID- 11901382 TI - Three-dimensional in vivo kinematics of the distal radioulnar joint in malunited distal radius fractures. AB - How malunion of the distal radius affects the kinematics of the distal radioulnar joint in vivo was evaluated. A novel computed tomography image-based technique was used to quantify radioulnar motion in both wrists of 9 patients who had unilateral malunited distal radius fractures. In the injured wrists dorsal angulation averaged 21 degrees +/- 6 degrees, radial inclination averaged 18 degrees +/- 5 degrees, and radial shortening averaged 21 +/- 3 mm. Clinically, the average range of motion of the injured wrists was 75 degrees +/- 25 degrees pronation and 73 degrees +/- 23 degrees supination. Kinematics of the radius during pronation and supination in the malunited forearms was indistinguishable from that in the uninjured forearms. In both the axis of rotation of the radius passed through the center of the ulnar head, although it shifted slightly ulnar and volar in supination and radial and dorsal during pronation. In contrast to previous in vitro biomechanical findings, there was no dorsovolar radial translation at the extremes of pronation or supination and no translation of the radius along the rotation axis. Soft tissues may play a larger role in limiting function than previously appreciated, and treatment may require correction of altered soft tissue structures as well as any abnormal bone anatomy. PMID- 11901383 TI - An anatomic reconstruction of the distal radioulnar ligaments for posttraumatic distal radioulnar joint instability. AB - Fourteen patients with posttraumatic distal radioulnar joint instability were treated with a reconstruction of the distal radioulnar ligaments. The technique is anatomically accurate, is reproducible, and requires less dissection than previously described techniques. Candidates for the procedure had joint instability and an irreparable triangular fibrocartilage complex. Ten patients had bidirectional instability. Two patients had a concurrent corrective osteotomy of the distal radius for a malunion. The procedure restored stability and relieved symptoms in 12 of 14 patients at 1 to 4 years' follow-up evaluation. One patient with a deficient sigmoid notch and one with ulnocarpal ligament injury did not achieve full stability. All patients attained near full pronation and supination. The procedure is an effective treatment for an unstable distal radioulnar joint when its articular surfaces are intact and the other wrist ligaments are functional, and it can be used in combination with a distal radius corrective osteotomy. PMID- 11901384 TI - Validation of the extensor carpi ulnaris groove as a predictor for the recognition of standard posteroanterior radiographs of the wrist. AB - This study was designed to establish the extensor carpi ulnaris groove (ECUG) as a reliable radiographic criterion for recognition of true neutral posteroanterior (PA) radiographs and to verify precise measurements of ulnar variance in a large cohort of patients. In 197 patients 197 wrists were evaluated with a series of radiographic views obtained during routine wrist arthrography. Posteroanterior views were taken in all patients at 90 degrees, 45 degrees, and 0 degrees arm abduction and in 171 patients at 90 degrees elbow flexion and 90 degrees arm abduction with full elbow extension. The ECUG position was classified according to its profile with the ulnar styloid as excellent, acceptable, or unacceptable. Ulnar variance was measured on all x-ray films. Statistical analysis included interobserver reliability with 100 x-ray films measured by 2 evaluators. The ECUG was excellent or acceptable in 100% of the x-rays on standard PA views (arm abducted 90 degrees ), 87% excellent or acceptable and 13% unacceptable on 45 degrees arm abduction views, and 23% excellent or acceptable and 77% unacceptable on 0 degrees arm abduction (adducted) views (all with the elbow flexed at 90 degrees ). With the arm at 90 degrees abduction and full elbow extension the ECUG was excellent or acceptable in 91% of cases. These results show that the ECUG is a reliable criterion to verify arm position during PA wrist radiography and therefore provides a standard for making treatment decisions. The need for repeat radiographs should be reduced. PMID- 11901385 TI - Periarterial sympathectomy in scleroderma patients: intermediate-term follow-up. AB - The use of periarterial sympathectomy (PAS) to manage chronic digital ischemia caused by scleroderma remains controversial. The duration of efficacy of PAS in managing scleroderma symptoms was evaluated by examining microvascular physiology, health-related quality of life, and patient satisfaction. Twenty-two patients (29 hands) with scleroderma, chronic vascular insufficiency, and a history of nonhealing digital ulcers unresponsive to nonsurgical treatment were evaluated. Isolated cold stress testing and laser Doppler fluxmetry measurements were used to evaluate microvascular perfusion before surgery and after a mean follow-up period of 31 months (range, 7-108 months). Digital temperatures did not change after surgery, but microvascular perfusion had increased significantly at a mean of 31 months in 22 hands. Health-related quality-of-life data were collected at a mean of 46 months (range, 11-108 months) after surgery. Subjective improvement (fewer ulcers, faster ulcer healing, and decreased pain) was reported for 18 of 22 patients (24 of 29 hands) at a mean follow-up period of 46 months; 6 patients remained ulcer free. Follow-up of scleroderma patients after PAS documented improved microvascular perfusion as evaluated by laser Doppler fluxmetry and by variable clinical results. PMID- 11901387 TI - Vascular endothelial growth factor promotion of neoangiogenesis in conventional nerve grafts. AB - The effect of vascular endothelial growth factor (VEGF) on angiogenesis and neovascularization of conventional nerve grafts was evaluated in 48 rabbits. A 2.5-cm segment of right sciatic nerve was removed and orthotopically repaired. This graft was wrapped in dialysis tubing to prevent vessel ingrowth from adjacent tissue. An osmotic pump delivered either VEGF (100 ng/h for 3 days) or control solution. Evaluation methods included angiography, vessel density, and nerve blood flow measurement at 3, 7, and 14 days. On day 3, 42% of the control nerves and 100% of VEGF-treated nerves had partial longitudinal neovascularization. Vessel density (0.84 +/- 0.22 vessel/mm(2)) and nerve blood flow [25.34 +/- 7.62 mL/(min.100 g)] in VEGF-treated nerves were significantly higher than control group values [0.23 +/- 0.13 vessel/mm(2) and 5.35 +/- 0.99 mL/(min.100 g)]. Progressive improvement in parameters was seen at 7 and 14 days. Vascular endothelial growth factor infusion accelerates longitudinal neoangiogenesis and shortens the nerve ischemic time to 3 days in this model. The revascularization of VEGF-treated conventional nerve grafts in a poorly vascularized bed is identical to that of grafts in a healthy bed. PMID- 11901386 TI - An advanced neuroprosthesis for restoration of hand and upper arm control using an implantable controller. AB - An advanced neuroprosthesis that provides control of grasp-release, forearm pronation, and elbow extension to persons with cervical level spinal cord injury is described. The neuroprosthesis includes implanted and external components. The implanted components are a 10-channel stimulator-telemeter, leads and electrodes, and a joint angle transducer; the external components are a control unit and transmitter-receiver coil. The system has completed preclinical testing and has been implanted fully in 3 persons and partially in 1 person, all with tetraplegia caused by spinal cord injury at C5 and C6. The minimum follow-up time for any system component is 16 months. All subjects had improvements in grasp strength, range of motion, and ability to grasp objects and increased independence in activities of daily living. Each subject became a regular user of the neuroprosthesis and is satisfied with it. The implanted components have not caused any medical complications. The operation of the electrodes and sensors has been stable. The data show that this advanced neuroprosthetic system is safe and can provide grasping and reaching ability to individuals with cervical level spinal cord injury. PMID- 11901388 TI - Assessment of nerve graft donor sites used for reconstruction of traumatic digital nerve defects. AB - Several donor nerve graft sites commonly are used when repairing segmental defects in sensory nerves distal to the wrist. The cross-sectional area and number of fascicles of donor nerves and specific digital nerve segments were investigated to provide guidelines for selection of nerve graft harvest sites according to defects encountered. Nerve segments were harvested from 10 fresh cadavers (20 upper extremities). Five sites of nerve graft were harvested: lateral and medial antebrachial cutaneous nerves (LABCN, MABCN), posterior and anterior interosseous nerves (PIN, AIN), and sural nerves. Four sites of typical segmental nerve defects were harvested in a zone protocol: common digital nerve (zone 4), proper digital nerve (zone 3), digital nerve distal to main dorsal branch at the metacarpophalangeal joint (zone 2), and digital nerve distal to trifurcation at fingertip (zone 1). Sural nerve is the most anatomically similar nerve graft for defects in zone 4 by cross-sectional area and number of fascicles. Lateral antebrachial cutaneous nerve is most appropriate for zone 2 and 3 injuries by both criteria. Fingertip grafts for zone 1 injuries displayed cross-sectional area similarity to PIN, AIN, and MABCN. With regard to number of fascicles, zone 1 digital nerves are most similar to LABCN donors. PMID- 11901389 TI - Chondroitin sulfate-coated polyhydroxyethyl methacrylate membrane prevents adhesion in full-thickness tendon tears of rabbits. AB - Polyhydroxyethyl methacrylate (pHEMA) membranes coated on one side with chondroitin sulfate (CS) were used to block adhesion physically and to reduce friction between healing flexor tendons and the surrounding tissue in rabbit forepaws after surgical repair. Digits with pHEMA-only, standard tendon sheath repair, and with no sheath repair were the controls. Over 12 weeks the CS-coated membranes were evaluated for joint flexion, adhesion limitation, and tendon healing progress. The membranes initially allowed for better flexion (ie, for 6 weeks), but their relative superior effectiveness faded afterward. Histology showed that adhesions were less severe and healing was better in the CS-pHEMA membranes at 3 and 6 weeks. If further studies determine precise amounts or thicknesses of CS coats that will maximize its healing properties, CS-pHEMA should prove useful in clinical settings in which restoration of tendon sheath integrity with a minimum of adhesions is not possible. PMID- 11901391 TI - Resection of the flexor digitorum superficialis reduces gliding resistance after zone II flexor digitorum profundus repair in vitro. AB - The effect of complete or partial resection of the flexor digitorum superficialis (FDS) tendon on the gliding resistance of the flexor digitorum profundus (FDP) tendon after FDP tendon repair was investigated. Twenty-four human FDP tendons were cut to 80% of their transverse section and repaired with a modified Kessler or a Massachusetts General Hospital augmented Becker suture technique. Gliding resistance was measured with the following constructs: intact state, sutured FDP tendon with FDS tendon intact, sutured tendon without FDS tendon, and sutured tendon with one slip of FDS tendon excised. After FDP repair the gliding resistance after modified Kessler repair increased 247% with FDS intact, 132% with one slip of FDS present, and 103% with FDS entirely removed. With a Becker repair, resistance increased 671% compared with normal with the FDS intact, 379% with one slip of the FDS, and 348% without the FDS tendon. Preserving the whole FDS resulted in a significantly larger increase in gliding resistance after FDP repair than did full or partial FDS removal, which were not significantly different from each other. These results suggest that the FDS tendon affects the gliding resistance under the pulley after FDP repair and that partial FDS excision may facilitate gliding of a bulky FDP repair. PMID- 11901390 TI - Long-term viability of articular cartilage after microsurgical whole-joint transplantation and immunosuppression with rapamycin, mycophenolate mofetil, and tacrolimus. AB - The survival or rejection of articular cartilage in heterotopic vascularized joint transplants in rats immunosuppressed with rapamycin (SDZ RAD), mycophenolate mofetil (MMF), and tacrolimus was evaluated histologically up to 1 year after surgery. The vascularized knee joint of an ACI donor rat was transplanted to the groin of a Lewis recipient rat. Nonimmunosuppressed allografts were evaluated after 6 weeks and 3 months, and immunosuppressed allografts and control isografts were evaluated after 6 weeks, 3 months, 6 months, and 1 year. No rejection was seen in the control isografts. All allografts without immunosuppression were rejected at 6 weeks and 3 months. Eighteen of 21 knee joint transplants immunosuppressed with SDZ RAD and 17 of 22 knee joint transplants immunosuppressed with MMF were rejected between 6 weeks and 1 year. SDZ RAD and MMF caused significant side effects including compromised wound healing and bone marrow suppression culminating in weight loss and death. Eighteen of 19 knee joints immunosuppressed with tacrolimus showed no signs of rejection up to 1 year after surgery. Long-term intermittent immunosuppression with tacrolimus was significantly superior to SDZ RAD and MMF in preventing rejection of the transplanted articular cartilage of a vascularized knee joint allograft up to 1 year after surgery. PMID- 11901392 TI - Comparison of nonsurgical treatment measures for de Quervain's disease of pregnancy and lactation. AB - de Quervain's disease of pregnancy and lactation is usually self-limited and responds well to nonsurgical treatment. We conducted a randomized prospective study on 19 wrists of 18 patients with de Quervain's disease who were either pregnant or breast-feeding. One group had a cortisone injection into the tendon sheath and the other group used thumb spica splints. All 9 patients with injections had complete pain relief with only one late recurrence. None of the patients with splints had complete pain relief; however, at the end of the lactation period, 8 had spontaneous resolution of symptoms and 1 received a cortisone injection. de Quervain's disease of pregnancy and lactation is self limited and can be treated successfully with cortisone injection. Splinting does not provide satisfactory pain relief. PMID- 11901394 TI - The rapid exchange grip strength test and the detection of submaximal grip effort. AB - This study assessed the reliability of the rapid exchange grip test for detecting submaximal grip effort, particularly evaluating its performance with motivated subjects with genuine hand weakness secondary to pain. Fifty normal participants performing with maximum effort then feigning hand weakness and 50 patients recovering from carpal tunnel surgery were studied. The results showed that the dynamic measure of grip strength equaled or exceeded the static measure in 28% of maximally performing participants (72% specificity), 58% of the carpal tunnel decompression patients (42% specificity), and 74% of participants giving submaximal grip effort (74% sensitivity). Sensitivities and specificities for other criteria of a positive test were also determined. Our findings suggest that the rapid exchange grip strength test cannot reliably detect voluntary submaximal effort. PMID- 11901393 TI - Variation in insertion of the abductor digiti minimi: an anatomic study. AB - We dissected 70 cadaver hands to examine the anatomy of the insertion of the abductor digiti minimi (ADM). The insertion had 2 forms. In 58 hands the insertion of the ADM was divided into 2 tendinous portions: one attached the base of the proximal phalanx (bone insertion) and the other attached the extensor aponeurosis (extensor insertion). In 12 hands the bone insertion existed solely. The mean length and width of the bone insertion were 10.6 +/- 3.1 and 4.0 +/- 0.9 mm and those of the extensor insertion were 18.5 +/- 4.0 and 2.1 +/- 1.1 mm, respectively. The mean proportion of the width of the extensor insertion to that of whole tendon was 31% (range, 0% to 50%). These observations show greater variation in the insertional anatomy of the ADM than reported previously. PMID- 11901395 TI - Evaluation of the Spanish version of the DASH and carpal tunnel syndrome health related quality-of-life instruments: cross-cultural adaptation process and reliability. AB - The methodologic requirements for cross-national use of specific health outcome instruments in hand surgery are presented. The Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire as a specific upper-extremity instrument and the Carpal Tunnel Syndrome (CTS) questionnaire as a specific disease instrument were adapted to Spanish for use in international studies. The adaptations were based on translation/back translation methodology. Meetings of translators, researchers, and patients were organized to produce successive versions. A study involving 50 people was carried out to consider the relative value (through a visual analog scale) of each response choice of the questionnaire items. Internal consistency and reproducibility of the Spanish version of both instruments were assessed by self-administering the questionnaires to 42 patients with the diagnosis of CTS on 2 different occasions 1 week apart. The average ratings of equivalence of the translated version with the original were high, regardless of the difficulty of translation. The adaptation process of the DASH and CTS instruments concluded with questionnaires conceptually equivalent to the original and with an acceptable level of reliability. PMID- 11901396 TI - Dupuytren's cord involving the septa of Legueu and Juvara: a case report. AB - A patient with Dupuytren's disease with involvement of the palmar fascial complex and digital contracture is described. A vertical cord had developed in the transverse ligament of the palmar aponeurosis fibers and the underlying septa of Legueu and Juvara. The cord was composed of a pretendinous band, transverse ligament of the palmar aponeurosis, and septum of Legueu and Juvara. The cord was attached deeply in the soft tissue confluence of the sagittal band, palmar plate, and interpalmar plate ligament. Involvement of the transverse ligament of the palmar aponeurosis and septa of Legueu and Juvara in Dupuytren's disease is rare. Understanding of the normal and pathologic fascial anatomy explains their simultaneous involvement and is necessary for complete ablation of the diseased tissue. PMID- 11901397 TI - High-pressure cement injection injury of the hand: a case report. AB - Modern equipment allows injection of substances at much higher pressures than previously. We describe a high-pressure cement injection injury to the hand and how its management differs from other injection injuries. This injury was treated by the established standard for this surgical wound: immediate debridement. The wound had the same mechanism, pathology, bacteriology, and treatment as other similar wounds. Prognosis after high-pressure injection injuries, however, also depends on the substance injected. Treatment for cement injection injuries differs because of the unique properties of cement. Immediate intervention is necessary for decompression and minimization of chemical burn. Removing the final few fragments of cement after they have hardened may decrease the number of debridements and soft tissue destruction. Serial x-rays can be used to guide debridements, but if serial x-ray films are not obtained, a final x-ray is mandatory to ensure removal of all cement. PMID- 11901398 TI - The intravenous foreign body: a report of 2 cases. AB - Two cases of metallic foreign body injury to the upper limb are described. In both cases the foreign body was clearly visible on x-rays, considered to be lodged in the soft tissues, but migrated to one of the large subcutaneous veins. One subsequently migrated to the heart; the other was removed from the peripheral vein. PMID- 11901399 TI - Bilateral bipartite lunate: a case report. AB - A rare case of bilateral bipartite lunate is reported. Radiographs showed that the lunate was divided into palmar and dorsal parts. Computed tomography and magnetic resonance imaging were used to differentiate this variant from lunate fracture. PMID- 11901400 TI - Southern California Society for Surgery of the Hand Award: best clinical contribution. PMID- 11901402 TI - Trephine bone grafting technique. PMID- 11901403 TI - Bridging a peripheral nerve defect using collagen filaments. PMID- 11901406 TI - Hereditary neuropathy with liability to pressure palsies. PMID- 11901405 TI - Hereditary neuropathy with liability to pressure palsies. PMID- 11901408 TI - Magnetic resonance imaging of triangular fibrocartilage complex lesions. PMID- 11901411 TI - Continence: the science, art, and charity of decency. PMID- 11901412 TI - Comments regarding January 2001 ostomy articles. PMID- 11901413 TI - Pouchitis: Part 1: Etiologies and risk factors. PMID- 11901414 TI - Quantitative and qualitative research: competition or parallel play? PMID- 11901415 TI - Managing thermal injuries within WOCN practice. AB - WOC nurses may be asked to manage small burn wounds or to follow up with patients who have residual wound care issues after recovering from a major burn injury. Aspects of care include identifying patients who warrant a higher level of burn care expertise, managing small wounds, recognizing the common complications of burns, and determining the needs of patients undergoing burn rehabilitation. Persons managing burn wounds will incorporate a variety of techniques to facilitate wound cleansing and dressing, pain management, psychological support, and minimization of complications. In addition, depending on the setting, extent, and nature of the wounds, the patient's abilities and rehabilitation requirements will need to be determined. The patient and his or her family need to be educated about the normal changes that follow a burn injury and how to manage these issues. PMID- 11901416 TI - Bacterial counts and types found on wound care supplies used in the home setting. AB - OBJECTIVE: Our objective was to determine the length of time that wound care supplies can be stored in the home setting before they become contaminated. With the rise in the numbers of antibiotic-resistant organisms, we wanted to try to determine if we were using safe practices when storing supplies in the home. DESIGN: This was a prospective, exploratory study. SETTING AND SUBJECTS: The setting was homes of persons who required wound care by a home health care agency in the Midwest. The types of wounds included postoperative wound complications, diabetic foot ulcer, pressure ulcer, venous stasis ulcer, chemotherapy extravasation, and radiation burn. Methods and Instruments: Cultures were collected from wound care supplies when the package was opened and on day 7 and day 14 with use of standard laboratory techniques. Two data collection tools were developed: a flow sheet used to record the length of time the package was opened and a tool that reported the laboratory's bacteriologic results to the investigator. RESULTS: Cultures were obtained from 3 sterile items, including one package of 4 x 4 gauze, one disposable suture set, and one bottle of (1/4)-inch packing gauze, before they were placed in the patient's home. These 3 baseline cultures were the control samples; they showed no growth. Cultures of the open packages of reusable supplies in the home began to show growth of different organisms by day 7. Cultures of the open packages left in the home for longer periods of time almost always showed a larger growth and variety of organisms that potentially could become problematic for a patient. CONCLUSION: This study identified the number and types of organisms found on wound care supplies used in the home setting on day 7 and day 14. Some colonies of organisms were large enough to be pathogenic to certain patients. This information was used to develop protocols for orientation of new staff, to perform skills testing of current staff, and to reinforce storage practices of wound care supplies left in the home. PMID- 11901417 TI - The development and pilot testing of a teaching booklet for oncology patients' self-assessment and perineal skin care. AB - Oncology patients often experience skin breakdown as a result of chemotherapy. Often the loss of skin integrity is neither identified nor treated until it becomes severe and painful. If patients were taught to identify early signs of breakdown and to report these symptoms to a health care provider, treatment for these conditions could begin at an early stage. Nursing staff identified the need to develop a booklet that would both instruct adult oncology patients concerning necessary perineal care following therapy and encourage them to partner with the nursing staff to provide this care through self-assessment and reporting. The focus of this article is the process of developing and pilot testing such a booklet by 27 female oncology patients undergoing peripheral stem cell transplantation. Participants were given a handheld mirror as an aid to assess perineal skin changes. Feedback from both staff and patients was very positive. As patients identified and reported perineal skin changes to the nursing staff, both patients and nurses implemented established perineal skin care protocols. This teaching booklet, The Perineal Skin Self-Assessment Guide, focuses on the partnering of nurses and patients to promote the involvement of patients in their own care. PMID- 11901418 TI - Interstitial cystitis: a guide to recognition, evaluation, and management for nurse practitioners. AB - Interstitial cystitis (IC) is a chronic disorder of unknown etiology that affects the lower urinary tract of up to 500,000 women and men in the United States. It is characterized by bladder and pelvic pain that varies from moderate discomfort to severe, debilitating pain and related lower urinary tract symptoms including nocturia, diurnal urinary frequency, and urgency. Because the symptoms of IC superficially resemble a urinary tract infection, it is often misdiagnosed and may remain so for months or even years. This article discusses the clinical manifestations of IC, including its differentiation from acute or recurring bacterial cystitis. Options for managing this significant and often debilitating voiding dysfunction are also discussed. PMID- 11901419 TI - Experiences of Swedish men and women 6 to 12 weeks after ostomy surgery. AB - PURPOSE: The purpose of this study was to describe patients' experiences, with a focus on body image, 6 to 12 weeks after a stoma operation. METHOD: Nine patients (8 of whom received preoperative teaching) and who were expected to have their colostomy, ileostomy, or urostomy for at least 6 months were interviewed about their feelings, attitudes, and life experiences after undergoing ostomy surgery. MAIN OUTCOME: Seven themes were identified: alienation from the body, altered body image, influences on sexual life, uncertainty, influences on social life, influences on sports and leisure activities, and physical problems. CONCLUSION: Despite the fact that 8 of the 9 subjects were given comprehensive preoperative teaching, all of the interviewees reported that the stoma influenced their daily life in many unexpected ways, and some expressed severe difficulty in coming to terms with the stoma. With a deeper understanding of patients' experiences, the WOC nurse and other health care staff can more effectively prepare and support patients in adapting to their new situation. PMID- 11901420 TI - Patient with peristomal pyoderma gangrenosum. PMID- 11901421 TI - Quantal release of free radicals during exocytosis of phagosomes. AB - Secretion of lysosomes and related organelles is important for immune system function. High-resolution membrane capacitance techniques were used to track changes in membrane area in single phagocytes during opsonized polystyrene bead uptake and release. Secretagogue stimulation of cells preloaded with beads resulted in immediate vesicle discharge, visualized as step increases in capacitance. The size of the increases were consistent with phagosome size. This hypothesis was confirmed by direct observation of dye release from bead containing phagosomes after secretagogue stimulation. Capacitance recordings of exocytosis were correlated with quantal free radical release, as determined by amperometry. Thus, phagosomes undergo regulated secretion in macrophages, one function of which may be to deliver sequestered free radicals to the extracellular space. PMID- 11901422 TI - Myosin IXb is a single-headed minus-end-directed processive motor. AB - Myosin is an actin-based molecular motor that constitutes a diverse superfamily. In contrast to conventional myosin, which binds to actin for only a short time during cross-bridge cycling, recent studies have demonstrated that class V myosin moves along actin filaments for a long distance without dissociating. This would make it suitable for supporting cargo movement in cells. Because myosin V has a two-headed structure with an expanded neck domain, it has been postulated to 'walk' along the 36-nm helical repeat of the actin filament, with one head attached to the actin and leading the other head to the neighbouring helical pitch. Here, we report that myosin IXb, a single-headed myosin, moves processively on actin filaments. Furthermore, we found that myosin IXb is a minus end-directed motor. In addition to class VI myosin, this is the first myosin superfamily member identified that moves in the reverse direction. The processive movement of the single-headed myosin IXb cannot be explained by a 'hand-over hand' mechanism. This suggests that an alternative mechanism must be operating for the processive movement of single-headed myosin IXb. PMID- 11901423 TI - The myosin converter domain modulates muscle performance. AB - Myosin is the molecular motor that powers muscle contraction as a result of conformational changes during its mechanochemical cycle. We demonstrate that the converter, a compact structural domain that differs in sequence between Drosophila melanogaster myosin isoforms, dramatically influences the kinetic properties of myosin and muscle fibres. Transgenic replacement of the converter in the fast indirect flight muscle with the converter from an embryonic muscle slowed muscle kinetics, forcing a compensatory reduction in wing beat frequency to sustain flight. Conversely, replacing the embryonic converter with the flight muscle converter sped up muscle kinetics and increased maximum power twofold, compared to flight muscles expressing the embryonic myosin isoform. The substitutions also dramatically influenced in vitro actin sliding velocity, suggesting that the converter modulates a rate-limiting step preceding cross bridge detachment. Our integrative analysis demonstrates that isoform-specific differences in the myosin converter allow different muscle types to meet their specific locomotion demands. PMID- 11901424 TI - Deregulated human Cdc14A phosphatase disrupts centrosome separation and chromosome segregation. AB - We show that human Cdc14A phosphatase interacts with interphase centrosomes, and that this interaction is independent of microtubules and Cdc14A phosphatase activity, but requires active nuclear export. Disrupting the nuclear export signal (NES) led to Cdc14A being localized in nucleoli, which in unperturbed cells selectively contain Cdc14B (ref. 1). Conditional overproduction of Cdc14A, but not its phosphatase-dead or NES-deficient mutants, or Cdc14B, resulted in premature centrosome splitting and formation of supernumerary mitotic spindles. In contrast, downregulation of endogenous Cdc14A by short inhibitory RNA duplexes (siRNA) induced mitotic defects including impaired centrosome separation and failure to undergo productive cytokinesis. Consequently, both overexpression and downregulation of Cdc14A caused aberrant chromosome partitioning into daughter cells. These results indicate that Cdc14A is a physiological regulator of the centrosome duplication cycle, which, when disrupted, can lead to genomic instability in mammalian cells. PMID- 11901425 TI - Aortic dissection: advances in imaging and endoluminal repair. PMID- 11901427 TI - Angioplasty of the innominate artery in 89 patients: experience over 19 years. AB - PURPOSE: To assess retrospectively the success of percutaneous transluminal angioplasty (PTA) in treating innominate artery stenoses and occlusions in a large series of patients with long-term follow-up results. METHODS: In symptomatic (upper limb claudication, transient ischemic attack, vertebrobasilar insufficiency) patients with high-degree (>60%) stenosis, innominate artery PTA was performed. Long-term follow-up was undertaken by blood pressure measurements on both arms as well as subclavian, right common carotid and right vertebral duplex scan. RESULTS: Between 1981 and 1999, the primary success rate of 89 innominate artery PTA (84 stenoses, 5 occlusions) was 96.4%. Complications included one left occipital lobe infarction (2%), two puncture-site thromboses (3%) and four transient ischemic attacks (6%). Two patients with restenosis were successfully treated with re-PTA. Cumulative primary patency was 98 +/- 2% at 6 months, 93 +/- 4% at 16-117 months; secondary patency was 100% at 6 months, 98 +/ 2% at 12-117 months. Sixty-one percent of the patients became symptomless, 32% improved, 7% showed no improvement. CONCLUSION: Angioplasty of the innominate artery has been proven to be safe and effective on a large series of patients. For innominate artery stenosis and short occlusion, PTA should be the treatment of choice. PMID- 11901426 TI - Gene therapy techniques for peripheral arterial disease. AB - Somatic gene therapy is the introduction of new genetic material into selective somatic cells with resulting therapeutic benefits. Vascular wall and, subsequently, cardiovascular diseases have become an interesting target for gene therapy studies. Arteries are an attractive target for gene therapy since vascular interventions, both open surgical and endovascular, are well suited for minimally invasive, easily monitored gene delivery. Promising therapeutic effects have been obtained in animal models in preventing post-angioplasty restenosis and vein graft thickening, as well as increasing blood flow and collateral development in ischemic limbs. First clinical trials suggest a beneficial effect of vascular endothelial growth factor in achieving therapeutic angiogenesis in chronic limb ischemia and the efficacy of decoy oligonucleotides to prevent infrainguinal vein graft stenosis. However, further studies are mandatory to clarify the safety issues, to develop better gene delivery vectors and delivery catheters, to improve transgene expression, as well as to find the most effective and safe treatment genes. PMID- 11901428 TI - Human thrombin injection for the percutaneous treatment of iatrogenic pseudoaneurysms. AB - PURPOSE: Thrombin injection is becoming well established for the percutaneous management of iatrogenic pseudoaneurysms. All the published series to date use bovine thrombin, and there have been reports of adverse immunologic effects following its use. Our study aimed to assess the efficacy of human thrombin injection for pseudoaneurysm occlusion. METHODS: Fourteen patients with iatrogenic pseudoaneurysms underwent a color Doppler ultrasound examination to assess their suitability for percutaneous human thrombin injection. Human thrombin 1000 IU was then injected into the pseudoaneurysm sac under sterile conditions and with ultrasound guidance. A further color Doppler ultrasound examination was performed 24 hr later to confirm occlusion. RESULTS: All 14 pseudoaneurysms were successfully occluded by human thrombin injection. In two cases a second injection of thrombin was required, but there were no other complications, and all pseudoaneurysms remained occluded at 24 hr. CONCLUSION: Ultrasound-guided human thrombin injection is simple to perform, effective and safe. We recommend that human thrombin becomes the agent of choice for percutaneous injection into iatrogenic pseudoaneurysms. PMID- 11901429 TI - Percutaneous imaging-guided access for the treatment of calculi in continent urinary reservoirs. AB - PURPOSE: To describe our long-term experience with percutaneous access to continent urinary reservoirs for calculus removal. PATIENTS AND METHODS: A retrospective study of 13 procedures in 10 patients was performed. In 2 of the 13 procedures, access and calculus removal was performed in a single session. In the other 11 procedures, initial access was obtained using ultrasonography, fluoroscopy, and/or computed tomography. The patients then returned at a later date for a second step where the access was dilated and the calculi were removed. RESULTS: Access was achieved successfully in all cases with no complications. At mean follow-up time of 13.6 months (range 1-94 months) one patient had died of complications unrelated to her continent urinary reservoir. Another patient had been placed on suppressive antibiotics for recurrent calculi. The remaining patients were stone free and without late complication. CONCLUSIONS: Percutaneous removal of reservoir calculi can be performed safely, avoiding potential injury to the continence valve mechanism by a direct cystoscopic approach. We propose a two-stage procedure using CT guidance for initial access as the preferred technique. PMID- 11901430 TI - Complications with outpatient angiography and interventional procedures. AB - PURPOSE: To prospectively identify the complications, and rates of complication, in outpatient angiography and interventional procedures. METHODS: There were 1050 consecutive patients, 646 men and 404 women, aged 17-89 years, with a total of 1239 procedures studied in a 2-year period, 1997 to 1999. RESULTS: There were 560 cases of aorto-femoral angiography, resulting in 124 complications (22%), with pain or hematoma in 110. There were 206 cases of neck and cerebral angiography, resulting in 51 complications (25%), with pain and hematoma in 34, transient ischemic attack in 2 and cerebrovascular accident in 1. There were 197 interventional procedures, with 177 being balloon dilatations, resulting in 68 complications (35%), with 2 having hematomas and 1 having hematoma/abscess requiring active treatment. There were 276 cases having various "other" procedures (e.g., renal angiography), resulting in 65 complications (24%), with pain and hematoma in 61. No procedure-related death occurred. Eighteen cases (1.5%) had significant complications, with contrast allergy in eight. CONCLUSION: Outpatient angiography and intervention are relatively safe, with low significant complication rates. PMID- 11901431 TI - A polyhydroxybutyrate biodegradable stent: preliminary experience in the rabbit. AB - PURPOSE: The lifelong persistence of foreign bodies within the arteries may contribute to restenosis. Thus, biodegradable devices might decrease recurrence rates. METHODS: Eleven polyhydroxybutyrate biodegradable stents and 13 tantalum stents were implanted into the iliac arteries of New Zealand white rabbits for up to 30 weeks. After killing the animals, the specimens were harvested, fixed in formalin, processed in paraffin, and stained. RESULTS: Polyhydroxybutyrate instigated intense inflammatory and proliferative reactions with an increase in collagen (2.4- to 8-fold vs native segments), thrombosis and in-stent lumen narrowing (375.5-606.6 mm vs 655.6 +/- 268.8 mm in native segments). The elastic membranes were destroyed in all specimens. The tantalum stents increased the in stent lumen progressively (769.7 +/- 366.6 mm vs 1309.9 +/- 695.3 mm), penetrated the external elastic membrane, and increased mural collagen content (6- to 8.6 fold vs native segments). Neither restenoses nor thromboses occurred. CONCLUSIONS: In the rabbit iliac artery, polyhydroxybutyrate stents caused intensive inflammatory vascular reactions which ban them from clinical use. PMID- 11901433 TI - Spontaneous rupture of hepatocellular carcinoma supplied by the right renal capsular artery treated by transcatheter arterial embolization. AB - We present a case of spontaneous rupture of hepatocellular carcinoma (HCC) with poor liver function which was treated by transcatheter arterial embolization (TAE). The patient's bilirubin value was 3.8 mg/dL. The tumor was fed by the right renal capsular artery according to selective arteriography. It was subsequently treated by TAE. With successful TAE, no hepatic failure was related to the treatment. We believe that if tumors are fed only by extrahepatic collateral vessels, TAE may be an effective treatment even in patients with poor liver function. PMID- 11901432 TI - Acute small bowel hemorrhage in three patients with end-stage renal disease: diagnosis and management by angiographic intervention. AB - Three patients who had undergone hemodialysis for end-stage renal disease, presented with acute small bowel hemorrhage, and were treated with superselective transcatheter arterial embolization via coaxial microcatheters. In all patients pre-procedure upper gastrointestinal (GI) endoscopy and colonoscopy had failed to demonstrate the source of the hemorrhage. Selective diagnostic angiography revealed frank extravasations of contrast from the small bowel arteries (one jejunal artery and two ileal arteries). After superselection of feeding arteries with a microcatheter, transcatheter embolization using Gelfoam and microcoils was performed in all three patients. Immediate hemostasis was achieved in all patients and the patients were discharged free from symptoms 3-5 days after embolization. No evidence of intestinal ischemia or infarction was noted, with the time from procedure to last follow-up ranging from 4 to 12 months. We conclude that superselective angiography is a valuable tool for diagnosing and treating acute small bowel hemorrhage in patients with end-stage renal disease when endoscopic evaluation has failed. PMID- 11901435 TI - Two fatal complications after parallel tracheal-esophageal stenting. AB - Two patients with malignant obstructions of both the trachea and esophagus underwent parallel stent placement with Gianturco-Rosch Z (GRZ) stents for palliation of symptoms. Fatal hemorrhage occurred in both patients 2 and 3 weeks after stent placement respectively. An autopsy performed on one of these patients demonstrated esophageal tissue necrosis and erosion with perforation of both the tracheal and esophageal walls at sites where the stent struts were in direct opposition, leading to bleeding from the esophageal venous plexus. GRZ stents have been successful in the treatment of both solitary tracheal and esophageal stenoses. However, parallel tracheal-esophageal stenting with GRZ stents places patients at high risk for complications due to the high radial force exerted by this particular stent and the minimal amount of intervening tissue between the two structures. PMID- 11901434 TI - Urokinase lysis for acute left subclavian artery thrombosis after placement of infusion catheter: report of two cases. AB - We present two cases of acute subclavian and/or axillary arterial occlusion after transaxillary catheterization with an implantable port for hepatic arterial infusion chemotherapy. They were successfully treated with thrombolytic therapy using intraarterial administration of urokinase without removal of the infusion catheter system. We consider that this treatment is suitable for managing acute thrombosis of the conduit artery after catheterization for hepatic arterial infusion chemotherapy. PMID- 11901436 TI - Restoration of liver function and portosystemic pressure gradient after TIPSS and late TIPSS occlusion. AB - TIPSS (transjugular intrahepatic portosystemic shunt) may be indicated to control bleeding from esophageal and gastric varicose veins, to reduce ascites, and to treat patients with Budd-Chiari syndrome and veno-occlusive disease. Numerous measures to improve the safety and methodology of the procedure have helped to increase the technical and clinical success. Follow-up of TIPSS patients has revealed shunt stenosis to occur more often in patients with preserved liver function (Child A, Child B). In addition, the extent of liver cirrhosis is the main factor that determines prognosis in the long term. Little is known about the effects of TIPSS with respect to portosystemic hemodynamics. This report deals with a cirrhotic patient who stopped drinking 7 months prior to admission. He received TIPSS to control ascites and recurrent esophageal bleeding. Two years later remarkable hypertrophy of the left liver lobe and shunt occlusion was observed. The portosystemic pressure gradient dropped from 24 mmHg before TIPSS to 11 mmHg and remained stable after shunt occlusion. The Child's B cirrhosis prior to TIPSS turned into Child's A cirrhosis and remained stable during the follow-up period of 32 months. This indicates that liver function of TIPSS patients may recover due to hypertrophy of the remaining non-cirrhotic liver tissue. In addition the hepatic hemodynamics may return to normal. In conclusion, TIPSS cannot cure cirrhosis but its progress may be halted if the cause can be removed. This may result in a normal portosystemic gradient, leading consequently to shunt occlusion. PMID- 11901437 TI - Percutaneous removal of a fractured endostent remnant from the portal vein. AB - We report the case of a liver transplant patient who developed a biliary stricture 3 years postoperatively which was treated with an endostent. During endoscopic removal, the stent fractured and a portion of it lodged itself within the intrahepatic portion of a portal vein branch. The endostent fragment was retrieved percutaneously using interventional radiology techniques. Risk factors for endostent fracture and migration as well as various percutaneous retrieval methods are reviewed in this article. PMID- 11901438 TI - Transvenous transjugular renal core biopsy with a redesigned biopsy set including a blunt-tipped needle. AB - A novel 19-gauge, blunt-tipped, side cutting single throw, 70-cm long transjugular needle, specifically designed for transvenous kidney biopsy, was used in seven patients with high risk for bleeding. A mean of 4 device-passes (3 6) per patient resulted in a satisfactory specimen for pathological diagnosis. Immediate post-biopsy nonenhanced CT was performed to evaluate for bleeding at the biopsy site. All patients were observed for 2 hr after the procedure. No clinically significant immediate or late complication was noted. PMID- 11901439 TI - Re: ureteroarterial fistula: endovascular repair with a stent-graft. PMID- 11901440 TI - Re: spontaneous rupture of an intercostal artery due to neurofibromatosis type I disease treated by percutaneous embolization. PMID- 11901442 TI - Vascular Dementia. PMID- 11901444 TI - Will there be room for the teaching of internal medicine in a university hospital? AB - To answer the question addressed, two working groups, one made of the staff of a University clinic, the other one composed of practising general internists, have discussed the assets and weaknesses of a University service of Internal Medicine for postgraduate training. The groups agreed on a number of points: patients' characteristics (complexity and co-morbidities), quality of teaching, method acquisition for clinical reasoning, as well as absence of exposure to ambulatory patients and of follow-up. The groups differed in their views related to the lack of training in psychiatry and psychosocial problems or to hospital dysfunctions. Opening of internal medicine to primary care appears to be necessary at the same time as individual qualities among the senior staff are to be developed, such as critical analysis and self-questioning. PMID- 11901445 TI - A single high dose of inhaled corticosteroids: a possible treatment of asthma exacerbations. AB - Recovery from an asthma exacerbation may take days or weeks even after the introduction of appropriate exacerbation therapy. However airway responsiveness and sputum eosinophils can be reduced within 6 hours by a single dose of inhaled corticosteroids. AIM: To determine if a single dose of 3200 microg of budesonide increases the rate of recovery from an asthma exacerbation. METHODS: Nineteen asthmatic subjects with an asthma exacerbation following withdrawal of inhaled corticosteroids were randomised to receive either usual care (doubling their dose of inhaled corticosteroids) plus placebo or usual care plus a single dose of 3200 microg of budesonide in a double-blind manner. Subjects monitored peak flow (PEF), symptoms, and beta agonist use daily for four weeks. The lowest PEF reading for each week was calculated as a percentage of the best peak flow value achieved in the recent past (PEF lowest % best). RESULTS: In the first week following exacerbation, PEF (lowest % best) was significantly greater in the budesonide group than in the placebo group (87.4 +/- 4.7 vs. 76.7 +/- 5.3; p = 0.029). However in the fourth week following exacerbation PEF was not significantly different (p = 0.728). The proportion of subjects who had a symptom free day during the first week was significantly higher in the budesonide group (p = 0.0012). CONCLUSION: A single high dose of inhaled corticosteroids added to usual exacerbation treatment might increase the rate of recovery from a mild exacerbation of asthma. PMID- 11901446 TI - Laparoscopic adrenalectomy: the new standard? AB - PRINCIPLES: Since 1994 we have been removing most non-malignant classified pathologies of the adrenal gland laparoscopically. Does this minimal invasive procedure involve advantages over the conventional approach? METHODS: Retrospective analysis of 22 all-consecutive laparoscopic adrenalectomies in 21 patients (10 women, 11 men, age 26-70 years, mean 43 years, 11 right, 9 left, one bilateral in MEN IIa syndrome). These procedures were performed between 1994 and 2001 transperitoneally in the lateral decubitus position, recently by use of the Ultracision device and once with a handport. These results are compared with 20 consecutive open transperitoneal unilateral adrenalectomies with similar pathologies (13 women, 7 men, age 28-77 years, median 51.5 years, 8 right, 12 left) carried out between 1988 and 1993. RESULTS: The mean operating times were 150 and 115 minutes with the laparoscopic and the open procedure respectively (p <0.011). On the other hand, mean hospital stay (6 versus 15 days, p <0.00001), intraoperative blood loss (200 versus 300 ml, p <0.04) and postoperative need for analgesics were significantly shorter or lower. Two out of the first five laparoscopic operations had to be converted into open adrenalectomy due to intraabdominal adhesions and a diaphragmatic injury with pneumothorax. In both groups three complications occurred (14% and 15%). CONCLUSION: Laparoscopic adrenalectomy is a safe, effective and useful procedure involving a shorter hospital stay, lower intraoperative blood loss and a lower postoperative analgesics requirement compared with the open approach. The laparoscopic approach is the procedure of choice for all benign adrenal pathologies. PMID- 11901447 TI - Gemcitabine-related pulmonary toxicity. AB - BACKGROUND: Gemcitabine is an increasingly used and generally well tolerated anticancer drug. Rarely, it leads to potentially fatal pulmonary toxicity. CASE DESCRIPTIONS AND RESULTS: We describe the clinical features of 5 patients with gemcitabine-related pulmonary toxicity. Due to early diagnosis, prompt discontinuation of the drug, and treatment with steroids, toxicity was reversible in all cases. CONCLUSION: Early recognition of gemcitabine-related pulmonary toxicity is mandatory. PMID- 11901448 TI - [The HYGEA Study]. PMID- 11901449 TI - [HYGEA (Hygiene in gastroenterology--endoscope reprocessing): Study on quality of reprocessing flexible endoscopes in hospitals and in the practice setting]. AB - The quality of reprocessing gastroscopes, colonoscopes and duodenoscopes in daily routine of 25 endoscopy departments in hospitals and 30 doctors with their own practices was evaluated by microbiological testing in the HYGEA interventional study. In 2 test periods, endoscopes ready for use in patients were found contaminated at high rates (period 1: 49 % of 152 endoscopes; period 2: 39 % of 154 endoscopes). Culture of bacterial fecal flora (E. coli, coliform enterobacteriaceae, enterococci) was interpreted indicating failure of cleaning procedure and disinfection of endoscopes. Detection of Pseudomonas spp. (especially P. aeruginosa) and other non-fermenting rods - indicating microbially insufficient final rinsing and incomplete drying of the endoscope or a contaminated flushing equipment for the air/water-channel - pointed out endoscope recontamination during reprocessing or afterwards. Cause for complaint was found in more than 50 % of endoscopy facilities tested (period 2: 5 in hospitals, 25 practices). Reprocessing endoscopes in fully automatic chemo-thermally decontaminating washer-disinfectors with disinfection of final rinsing water led to much better results than manual or semi-automatic procedures (failure rate of endoscopy facilities in period 2 : 3 of 28 with fully automatic, 8 of 12 with manual, 9 of 15 with semi-automatic reprocessing). The study results give evidence for the following recommendations: 1. Manual brushing of all accessible endoscope channels has to be performed even before further automatic reprocessing; 2. For final endoscope rinsing, water or aqua dest. should only be used disinfected or sterile-filtered; 3. Endoscopes have to be dried thoroughly using compressed air prior to storage; 4. Bottle and tube for air/water-channel flushing have to be reprocessed daily by disinfection or sterilization, and in use, the bottle have to be filled exclusively with sterile water. The HYGEA study shows that microbiological testing of endoscopes is useful for detection of insufficient reprocessing and should be performed for quality assurance in doctors' practices, too. The study put recommendations for reprocessing procedures in more concrete terms. PMID- 11901450 TI - Non-invasive evaluation of activity in inflammatory bowel disease by power Doppler sonography. AB - BACKGROUND: To study the vascularization in the diseased bowel wall by power Doppler sonography in patients with inflammatory bowel disease. PATIENTS AND METHODS: The diseased bowel wall was investigated in 99 patients with inflammatory bowel disease (60 patients with Crohn's disease and 39 patients with ulcerative colitis) either with active disease or in remission by B-mode and power Doppler sonography. Disease activity was determined by clinical indices. Twenty healthy age and sex matched individuals served as controls. RESULTS: Bowel wall was thickened in active Crohn's disease (mean 7 mm, range 4-14) and ulcerative colitis (mean 5 mm, range 2-15) as compared to healthy controls (mean 2 mm, range 1-3), p < 0.001. In contrast to healthy controls blood vessels were detected in the bowel wall in 100 % of patients with active Crohn's disease and 91 % with active ulcerative colitis. Vascularization was significant decreased in patients with quiescent versus active disease in ulcerative colitis (p < 0.05), while in Crohn's disease there was no significance between active and remission phase. CONCLUSIONS: Thickened and hypervascularized bowel wall are characteristic findings in inflammatory bowel disease. A combination of B-mode and power Doppler sonography offers an additional noninvasive procedure for the determination of activity in patients with inflammatory bowel disease. PMID- 11901451 TI - Prevalence of functional bowel disorders and related health care seeking: a population-based study. AB - OBJECTIVES: There are few population-based studies on prevalence of functional bowel disorders (FBD) and related health care seeking. The aim of the present study was to evaluate the prevalence of FBD in a population-based sample and to assess FBD-related health care seeking and medication in Germany. MATERIAL: Cross sectional study, based on an age- and sex-stratified random sample of 2,400 subjects aged 21-80 years in Dusseldorf, Germany (about 500,000 population). Assessment was performed using a postal written questionnaire. METHODS: Prevalence of gastrointestinal pain or discomfort in the past 12 months was assessed, in particular, lower abdominal pain and irritable bowel syndrome (IBS). Furthermore, health care seeking and medication (prescribed and over-the-counter) due to FBD was assessed. Multiple logistic regression (survey estimated) was performed to evaluate associations of FBD with age, sex, and the socioeconomic status (SES). RESULTS: 1,281 subjects (53.4 %) were analyzed. Standardized prevalences were 22.6 % (95 %-CI: 20.3 - 25.1 %) for lower abdominal pain and 12.5 % (10.7-14.5 %) for IBS. Both lower abdominal pain and IBS were significantly less frequent in the older population compared to younger subjects. No significant differences were found for gender and SES. Among subjects with lower abdominal pain and IBS, 55.1 % and 49.3 % reported health care seeking due to their GI disorders, and 63.9 % and 56.2 % reported use of medication, respectively. CONCLUSIONS: A high prevalence of functional bowel disorders was found in this population-based study in Germany. Only about half of the subjects reported health care seeking due to their bowel disorders. Self-medication with over-the-counter agents was frequently performed. PMID- 11901452 TI - [Cystic lymphangioma with endosalpingiosis as a rare cause of gastrointestinal bleeding]. AB - We report on a 31-year-old woman with an iron-deficiency anemia and positive Guaiac test, existing for more than 2 years. Despite an extensive gastrointestinal examination a bleeding source could not be found. Also angiography of the abdominal arteries with injection of heparin as provocative protocol was normal. Enteroclysis showed a remarkable finding which led to laparotomy. A cystic lymphangioma with endolymphatic endosalpingiosis as the bleeding source was identified and removed. Lymphangioma is a rare benign soft tissue tumor which usually appears during the first 2 years of life. Only in some cases the lymphangioma is intraabdominal and there can be different, no specific symptoms. Endosalpingiosis is also a rare benign disease, it consists of scattered epithelium of the Fallopian tube. It is the first case to our knowledge, in which the appearance of the endosalpingiosis in a lymphangioma is described. PMID- 11901454 TI - [Crohn's disease and stromal sarcoma of the small bowel--a rare coincidence]. AB - We report the case of a 29-year-old female who suffers for more than 13 years from Crohn's disease. A small bowel resection had to be performed because of a complicated and insufficiently controlled course of the disease with beginning subileus. At laparotomy a leiomyosarcoma was found. This case represents the forth case in the literature of an association between chronic inflammatory bowel disease and malign mesenchymal tumors. Tumors of epithelial and lymphoreticular origin are known as a much more common complication in Crohn's disease and especially ulcerative colitis. An exact histological characterization of these sarcomas is important to define the prognosis of the disease. If recurrent subileus is poorly controlled in Crohn's disease stromal tumors with their frequent intraluminal growth have to be considered as part of the differential diagnosis. PMID- 11901453 TI - [Intussusception after Billroth II procedure--an uncommon cause of an upper gastrointestinal ileus]. AB - We present a case of a 73-year-old male patient with Korsakow's disease who was admitted with upper gastrointestinal bleeding and recurrent vomiting. He had received partial gastric resection with Billroth II reconstruction 39 years before for recurrent ulcer disease. At gastroscopy erosive gastritis with no active bleeding and a structure-resembling necrotic mucosa suspicious for intussuscepted small bowel was seen. At exploratory laparotomy jejunogastric intussusception of 50 cm of small bowel through Braun's enteroanastomosis into the gastric remnant was found. After reposition the bowel recovered well and resection was unnecessary. As prophylaxis the bowel was partially attached by sutures in terms of a partial Noble's Operation. The patient's recovery was uneventful after surgery. Jejunogastric intussusception is a rare cause of upper gastrointestinal bleeding and ileus. PMID- 11901455 TI - [Probiotics in gastroenterology]. AB - Probiotics are defined as live microorganisms of human origin. Their use may favorably influence human health and ameliorate or prevent certain diseases. Prebiotics are non-digestible foodstuffs (fiber, oligofructans - "colonic foods"), which enter the colon and are metabolized by the probiotics. Probiotics should fulfill the following criteria: Phenotypic and genotypic classification, no pathogenic properties, human origin, application in the living state, resistance to gastric acid and bile, ability to adhere to colonocytes, ability to colonize the gut, clinically proved favorable health-effect, and safety. Experimental and clinical studies supplied evidence of the possible use of probiotics in the following diseases: Traveler's diarrhea, antibiotic-associated diarrhea, relapsing Clostridium difficile colitis, infantile diarrhea, rotavirus enteritis, inflammatory bowel disease, irritable bowel syndrome, colon cancer, peritonitis, acute pancreatitis, and diarrhea associated with HIV infection. Probiotics displayed the following effects in these studies: Involvement in production of essential nutrients of the colonic mucosa, beneficial effect on intestinal immunity, recovery of the disturbed gut mucosal barrier and prevention of microbial translocation, elimination of toxins and eradication of microbial pathogens, production of steroids from cholesterol and reduction of its pool in circulation, participation in regulation of intestinal functions, reduced incidence of chemically induced colon tumors in rodents. Probiotics open new therapeutic modalities in a number of diseases and it may be expected that their importance will increase with growing knowledge and experience. PMID- 11901456 TI - [Preoperative short-time radiotherapy in resectable rectal cancer: A "new standard" coming up?]. PMID- 11901457 TI - [NOD2: A functional and positional candidate gene in Crohn's disease]. PMID- 11901458 TI - Arthroscopically-assisted reduction of intra-articular fractures and soft tissue management of distal radius. AB - Arthroscopy was used to help to reduce intra-articular fractures of the distal radius and treat soft tissue injuries in 33 acute cases. The fractures were treated by reduction under arthroscopic control and percutaneous fixation either with or without external fixation. The triangular fibrocartilage complex (TFCC) was torn in 18 of 33 patients (54%). All tears were peripheral and were repaired with arthroscopic procedures. Scapholunate (SL) ligament injuries prevailed in six (18%) patients; most of them exhibited instability in the SL joint. They received SL ligament debrided and transfixed with K-wires. Four (12%) of the patients suffered lunotriquetral (LT) ligament injuries; three of them also received transfixation with K-wires. Six (18%) of the patients exhibited chondral fractures. All fractures healed without measurable incongruity of joint surface and at follow-up (24 to 36 months), 11 patients displayed excellent results and 22 patients displayed good results according to the Mayo modified wrist score. PMID- 11901459 TI - Conservative treatment of redisplaced Colles' fractures in elderly patients older than 60 years old - anatomical and functional outcome. AB - A retrospective study was carried out to compare the functional outcome in two groups of patients who were 60 years old or older. One group had Colles' fractures, which had been reduced and did not redisplace. The other group's fractures redisplaced and were treated conservatively. Of the 11 patients in the first group, 82% had excellent or good functional outcome, which was not significantly different when compared with the second group of 25 patients with 68% having excellent or good functional outcome. PMID- 11901460 TI - Lunate plasty for Kienbock's disease: use of a pedicled vascularised radial bone graft combined with shortening of the capitate and radius. AB - We treated eight patients with Kienbock's disease (two patients each with stage 1, 2, 3a and 3b disease by Lichtman's classification) by removing a pedicled, vascularised bone segment from the dorsal aspect of the distal radius and engrafting it into the lunate. Additional shortening of the radius was performed in patients with the ulna-minus or null variant. Shortening of the capitate and capito-hamate fusion were also performed in patients with stage 3 disease. All patients were relieved of their wrist pain at rest and during movement, and the mean grip strength increased from 37% of that in the contralateral hand before surgery to 80% after surgery. The mean post-operative range of motion in the affected wrist was 92% of that in the opposite wrist in patients with stage 1 and 2 disease, and 53% in patients with stage 3 disease. Post-operative assessment revealed that four patients had excellent results, three had good results, and one had a fair result. PMID- 11901461 TI - Magnetic resonance angiography of the forearm and hand in children. AB - A two-dimensional time-of-flight magnetic resonance angiography was performed in 29 arms in 25 pediatric patients with congenital hand abnormalities, whose average age was three years and eight months. Venous structures were eliminated with presaturation technique and the remaining arterial system was evaluated. Magnetic resonance angiography demonstrated major arteries in the forearm well but not their branches. Even the biggest branch of the artery sometimes could not be detected. Magnetic resonance angiography and Allen test were consistent in determining patency of the palmar arch in 62% of the cases but the sensitivity was only 28%. Our experience showed that non-invasive, convenient, two dimensional time-of-flight magnetic resonance angiography was useful for detecting continuity and spatial localisation of the major arteries in a child's forearm. However, it was not a complete alternative to conventional angiography and was unsatisfactory in delineating the vascular anatomy in the hand. PMID- 11901462 TI - Prevention of the parrot beak deformity in fingertip injuries. AB - The nail is supported by the dorsal tuft of the terminal phalanx. Following distal fingertip trauma, varying amounts of nail support may be lost resulting in the nail curving palmarwards. This curvature is dependent upon the degree of bony loss, the amount of remaining nail bed and the degree of scar contracture at the hyponychial-pulp interface. The parrot beak or hooked nail deformity is most commonly caused by tight closure of a fingertip amputation and excessive palmar tension at the hyponychial-pulp suture line. A simple technique using a hypodermic needle, that eliminates tension from the suture line at the fingertip pulp-hyponychial interface and prevents hooked nail deformity is described. This allows healing to take place without any palmarwards pull of the scar and a subsequent parrot beak deformity. PMID- 11901463 TI - Pathogenesis of the gradually elongated nerve. AB - The rabbits' sciatic nerves were lengthened by 30 mm in increments of 2.0 mm/day (2-mm group) and 4.0 mm/day (4-mm group). In the 2-mm group, the phosphorylated neurofilament (p-NF) immunoreactivity of axons was similar to that of the control group. However, in the 4-mm group, number of p-NF positive axons decreased. The number of p-NF positive cells at the seventh lumbar dorsal root ganglion cells of the 4-mm group was significantly larger than that of the control group. Abnormal p-NF immunoreactivity in the 4-mm group suggested an impairment of the axonal flow. Leakage of Evans blue-albumin into the endoneurial space, which meant destruction of the blood-nerve barrier function, was clearly evident in the 4-mm group, but minor in the 2-mm group. A speed of 2.0 mm/day, therefore, appears to be critical for safe nerve elongation in this model. PMID- 11901464 TI - The effect of aspirin and prostaglandin E(1) on the patency of microvascular anastomosis in the rats. AB - The purpose of this study was to evaluate the effect of oral prostaglandin E(1)(PGE(1)) on the patency of the microvascular anastomosis of the carotid artery in rat. A total of 48 rats were used, and divided into three groups. The first group (A) was used as a control with no medical agent being used after anastomosis, the second group (B) was medicated with aspirin, and the third group (C) was medicated with oral PGE(1). In each group, four rats were sacrificed serially on every post-operative 3, 5, 10 and 15 days after arterial anastomosis. Patency and histologic evaluations at the anastomotic site were observed. The results revealed that the PGE(1) therapied group showed highest patency rate (100%), lesser formation of mural thrombosis, and also minimal changes in the intimal hyperplasia and medial fibrosis. From these findings, we could conclude that PGE(1) has superior effect on maintaining the patency after microvascular surgery. PMID- 11901465 TI - The evaluation of pre- and post-operative classification system for cubital tunnel syndrome. AB - Pre- and post-operative classification systems for cubital tunnel syndrome are discussed in this study. Although there are several pre-operative evaluations, a suitable system should be easy to categorise, simple, reliable and reproducible. McGowan's grading system has been widely used for these reasons; however, grade II included complicated cases. Accordingly, McGowan's grade II is divided into two groups, such as grade II-A (relatively good prognostic group) and group II-B (relatively poor prognostic group). As far as the post-operative classification system is concerned, 4 grade (excellent, good, fair, and poor) classification is the most useful and easy to understand. PMID- 11901466 TI - Membranous tissue under the flexor carpi ulnaris muscle as a cause of cubital tunnel syndrome. AB - Membranous tissue under the flexor carpi ulnaris (FCU), hypertrophy and fascia of the FCU were examined in 90 patients with cubital tunnel syndrome; it (submuscular membrane) was found in all patients, excluding five whose data were incomplete. Impressions on the nerve under the FCU were noted in 75 patients. In 43 patients, the submuscular membrane was judged to be the cause of neural compression. These 75 patients included 60 and 15 cases whose fascia of the FCU were judged to the thick and thin, respectively. Among the 58 with a thick submuscular membrane, 56 patients had an impression under the membrane. Fifteen patients had a flat impression on the nerve in the proximal compartment of the FCU, which was formed by ligamentous and bony floor and thick fascia of the FCU. It is believed that increased pressure in the compartment is the cause of a flat impression. Slight compression of the nerve under the FCU with compressions in proximal and distal part of the nerve can cause symptoms of the nerve compression by multiple crush effect. PMID- 11901467 TI - Cubital tunnel release with lift-type endoscopic surgery. AB - A new technique of endoscopic release of the ulnar nerve at the elbow was designed for cubital tunnel syndrome. After three 5 mm incisions were made along the line of the ulnar nerve, the skin and subcutaneous tissue were lifted up using a fine tape to produce a provisional space. An endoscope was inserted through the one incision, and the constricting ligaments and fascia were released using a retrograde knife inserted through the other incision under endoscopic vision. Eight patients were treated using this method, and successful results were achieved. No neurovascular complications occurred, and all pre-operative complaints were resolved within three weeks. Our surgical series indicated an earlier return to work and daily activity due to early healing of incisions and minimal post-operative pain. PMID- 11901468 TI - Ulnar groove plasty for friction neuropathy at the elbow. AB - In this paper, we report on a new operation for ulnar neuropathy caused by friction at the elbow. The operation consists of ulnar groove plasty proximal to the cubital tunnel. The ulnar nerve is replaced into this reconstructed groove. Patients are relieved of discomfort, and motor and sensory functions are recovered. The nerve is kept stable throughout the full range of elbow motion and showed neither irritation nor adhesion. Friction ulnar neuropathy is traditionally treated by anterior transpositon or medial epicondylectomy. The ulnar nerve may become entrapped in scar tissue after these operations. We believe that this anatomical position is optimum for the nerve and that this procedure is essential for treatment of friction neuropathy. PMID- 11901469 TI - Gene therapy for rheumatoid arthritis. AB - Advances in understanding the biology of rheumatoid arthritis (RA) have opened new therapeutic avenues. One of these, gene therapy, involves the delivery to patients of genes encoding anti-arthritic proteins. This approach has shown efficacy in animal models of RA, and the first human, phase I trial has just been successfully completed. Hand surgery featured prominently in this pioneering study, as a potentially anti-arthritic gene encoding the interleukin-1 receptor antagonist was transferred to the metacarpophalangeal joints of subjects with RA one week before total joint arthroplasty. This study has confirmed that it is possible to transfer genes safely to human joints. It should pave the way for additional application of gene therapy to arthritis and other orthopaedic conditions. PMID- 11901470 TI - Congenital true complete symphalangism of all proximal interphalangeal joints of hands with carpal anomalies: a case report. AB - Symphalangism is a rare condition which manifests in either PIP or DIP joint congenital fusion. Symphalangism may be with some other skeletal deformities. In our case, all PIP joints of both hands were fused with bilateral hypoplasia of carpal bones and Minaar type III lunatotriquetral coalition congenitally. No motion was detected in PIP joints with absence of cutaneous creases over all PIP joints. Radiologically, carpal hypoplasia and lunatotriquetral coalition were seen in all fingers with the absence of PIP joints. The patient's skeletal survey revealed no other pathology. He had no complaints related to his hands. So, regular follow-up was recommended. This pathology was presented as rarely observed although clinical problem is usually not so much. PMID- 11901471 TI - Recurrent dislocation of the ECU tendon in a golf player: release of the extensor retinaculum and partial resection of the ulno-dorsal ridge of the ulnar head. AB - A 20-year-old golf player who developed recurrent dislocation of the extensor carpi ulnaris (ECU) tendon is reported. The patient was successfully treated by release of the extensor retinaculum and partial resection of the ulno-dorsal ridge of the ulnar head, because the methods caused no redislocation and required only a brief period of rehabilitation. PMID- 11901472 TI - The Hair-Thread-Tourniquet Syndrome - report of an unusual presentation of this rare condition. AB - The Hair-Thread-Tourniquet syndrome is a rare disorder with merely 17 cases of finger involvement reported so far. This is a report of one such case of accidental digital constriction, which presented as late as three months after injury. This is the longest reported duration between injury and detection. The constriction band was in the form of an eschar lying in an epithelialised crease. Treatment involved excision of the crease followed by multiple z-plasties to close the resulting defect. This is the first case in English literature to report epithelialisation of the crease and consequently the first case to require excision of the band followed by z-plasty. PMID- 11901473 TI - Dupuytren's disease of the wrist. AB - The authors present the case of a 47-year-old female who presents with bilateral Dupuytren's disease of the wrist. To date, three English language papers reporting on the occurrence of Dupuytren's disease at the wrist have been presented. These reports represent cases in males with unilateral involvement of their wrists where there has been primary Dupuytren's disease of the palm apparently spreading to the wrist in continuity. Our case demonstrates isolated bilateral Dupuytren's disease of the wrist in a female. The authors feel that this clinical presentation is rare and that consideration should be given to the diagnosis of Dupuytren's disease when there is a painful lesion of the soft tissues adjacent to the wrist. The presence of bilateral lesions at the wrist should further heighten the suspicion of Dupuytren's disease even in the absence of associated disease in the palm or fingers. A discussion of the previous reports is included. PMID- 11901474 TI - Metastasis of an adenocarcinoma of the stomach to the 4th metacarpal bone. AB - Metastatic tumours of the hand are uncommon. The majority of these tumours affect the phalanges and the primary tumours are usually bronchogenic in origin, with breast and kidney tumours next in frequency. Metastatic gastrointestinal to the hand is rare and usually from the colon. We report a case of poorly differentiated adenocarcinoma of the stomach antrum presenting with a metastatic lesion to the right 4th metacarpal bone. A review of the literature is included. PMID- 11901475 TI - The bacterial cell wall as a source of antibacterial targets. AB - This review focuses on target-based approaches for developing new chemical classes of antibacterial agents aimed at the bacterial cell wall. The clinical success of antibiotics such as beta-lactams and glycopeptides validates this chemotherapeutic strategy and emerging resistance to these agents warrants the development of new antibacterial drugs. Understanding the mechanism of action and resistance to beta-lactams and glycopeptides at a molecular level has supported the development of new agents that prevent transpeptidation and transglycosylation reactions of peptidoglycan polymerisation. The enzymes involved in the synthesis of the peptidoglycan structural unit have also been targeted for antibacterial discovery. The influence of bacterial genetics and genomics, structural biology, assay development and the properties of known inhibitors on these approaches will be discussed in the context of drug discovery. PMID- 11901476 TI - Emerging targets in the AKT pathway for treatment of androgen-independent prostatic adenocarcinoma. AB - Prostatic adenocarcinoma (CaP) is the most common, non-cutaneous malignancy and the second-leading cause of cancer death in men. The disease has two distinct phases: the androgen-dependent phase, which can be treated effectively with androgen ablation therapies, and the androgen-independent phase, for which there is no effective life-prolonging therapy. An estimated 32,000 men will die this year from androgen-independent, metastatic CaP. Efforts to understand the metastatic progression of CaP and the emergence of androgen-independent disease have begun to illuminate the molecular events involved. Recent work suggests that CaP progression to androgen-independent, metastatic disease involves a dampened apoptotic response, a release from the cell cycle block that initially follows androgen withdrawal and a shift from dependence on paracrine-derived growth and survival factors to autonomous production of these key proteins. Functional loss of the tumour suppressor phosphatase and tensin homologue deleted on chromosome ten (PTEN) and subsequent activation of the AKT pathway, have been prominently implicated in the progression of CaP to androgen-independence. Activation of the AKT pathway can suppress the apoptotic response, undermine cell cycle control and selectively enhance the production of key growth and survival factors. Though many proteins and intracellular signalling pathways can influence these biological processes, activation of the AKT pathway may be a particularly potent signal involved in CaP progression to androgen-independence and therefore presents a series of potential targets for therapy of advanced androgen independent CaP. PMID- 11901477 TI - The Cytokine Odyssey 2001, a joint meeting of the Society of Leukocyte Biology and the International Cytokine Society. 8-11 November 2001, Maui, Hawaii, USA. AB - The Cytokine Odyssey 2001 was held at the Outrigger Wailea Resort in Maui, Hawaii, USA. The meeting, jointly sponsored by the International Cytokine Society (ICS, 9th Annual Meeting) and the Society of Leukocyte Biology (SLB, 35th Annual Meeting), was organised by Carl Ware (Chair) from the La Jolla Institute for Allergy and Immunology (La Jolla, USA) and Thomas Hamilton (Co-Chair) from the Cleveland Clinic Foundation (Cleveland, USA). This international conference was designed to bring together leading investigators in molecular and cellular biology, physiology and genetics, interested in cytokines and cells of the immune system. This forum was aimed to assess the impact of this expanding science on new approaches to disease intervention [1]. PMID- 11901478 TI - New discoveries in prostate cancer biology and treatment. 5-9 December 2001, Naples, Florida, USA. AB - Androgen independence and bone metastasis are lethal complications in patients with advanced prostate cancer. Presently, there is no cure for patients with androgen-independent prostate cancer. In order to develop more effective therapies for this disease, the molecular events involved in the development of androgen independence and bone metastasis must be elucidated and then targeted by therapeutic agents. Several studies presented at a recent conference on prostate cancer sponsored by the American Association for Cancer Research (AACR) provided evidence that prostate cancer metastasis to bone is mediated by the prostate cancer cell expression of molecules that allow the cells to invade, grow in and stimulate cells in the bone microenvironment resulting in an osteoblastic reaction. Androgen independence was reportedly mediated by an increased expression of survival genes following androgen ablation therapies and several molecular mechanisms involved in genetic instability. Treatment strategies are being designed to target some of the molecular events involved in androgen independence and bone metastasis. Targeting these molecular events with combinational therapies will hopefully delay the progression to androgen independence in patients with early stage disease, suppress the growth of androgen-independent cells in patients with advanced disease and enhance the chemosensitivity of androgen-independent cells. PMID- 11901479 TI - Emerging therapeutic targets in tuberculosis: post-genomic era. AB - Every minute, somewhere in the world four people die from tuberculosis (TB), yet it has been nearly 40 years since a novel drug was introduced to treat this disease. The ever increasing number of TB cases together with the advent of multi drug resistant (MDR) TB, has stimulated the search for novel anti-TB agents. An array of novel drug targets is provided by the mycobacterial cell wall, whose integrity is essential for bacterial viability. Over the years researchers have identified potential drug targets that are associated with the synthesis of various cell wall constituents. This classic approach, together with the unravelling of the Mycobacterium tuberculosis genome sequence, has placed TB drug research in an unprecedented position. An entire new set of genetic and bioinformatic tools for probing potential drug targets is now available. As therapies using first-line drugs like isoniazid (INH) or rifampin in combination with second-line drugs, like ethambutol (EMB) still continues, a number of substituted fluoroquinolones are being considered as the new generation of anti TB drugs for their favourable pharmacokinetic profile and excellent oral bioavailability. In this review, the future of anti-TB drugs is discussed with reflection on the structure and biosynthesis of cell wall constituents that are potential drug targets. The importance and relevance of the M. tuberculosis genome sequence for the development of novel anti-TB drugs, have also been underscored. PMID- 11901480 TI - Novel therapeutic targets in osteoporosis. AB - Osteoporosis is a common condition in which significant bone loss occurs resulting in an increased risk of sustaining fractures. Several licensed therapies are available to treat this condition, which suffer from several disadvantages including limited efficacy, high cost and poor long-term patient adherence as a consequence of significant side effects and inconvenient methods of administration. A wide range of therapeutic targets have been developed to provide a basis for developing newer therapies which overcome these limitations. These can be subdivided into those that are primarily directed towards inhibiting osteoclast-dependent bone resorption and those that stimulate osteoblastic bone formation. Targets can be grouped as follows: systemic factors such as steroid and peptide hormones; local factors produced in bone involved in osteoblast and osteoclastic regulation; and cellular targets such as cell membrane receptors and attachment proteins, cellular enzymes and nuclear transcription factors. To date, only a small proportion of these targets have yielded novel compounds to have entered clinical trials. However, it is anticipated that these will provide the basis for significant numbers of new therapies for osteoporosis in the foreseeable future. PMID- 11901481 TI - Neuroendocrine influence on thymic haematopoiesis via the reticulo-epithelial cellular network. AB - The thymus provides an optimal cellular and humoral microenvironment for a cell line committed differentiation of haematopoietic stem cells. The immigration process requires the secretion of at least one peptide, called thymotaxin, by cells of the reticulo-epithelial (RE) network of the thymic stromal cellular microenvironment. The thymic RE cells are functionally specialised based on their intrathymic location and this differentiation is modulated by various interaction signals of differentiating Thymocytes and other nonlymphatic, haematopoietic stem cells. The subcapsular, endocrine, RE cell layer is comprised of cells filled with periodic acid Shiff's-positive granules, which also express A2B5/TE4 cell surface antigens and MHC Class I (HLA A, B, C) molecules. Thymic nurse cells also produce thymosins beta 3 and beta 4 and display a neuroendocrine cell specific immunophenotype (IP): Thy-1+, A2B5+, TT+, TE4+, UJ13/A+, UJ127.11+, UJ167.11+, UJ181.4+ and presence of common leukocyte antigen (CLA+). Cortical RE cells express a surface antigen, gp200-MR6, which plays a significant role of thymocyte differentiation. Medullar RE cells display MHC Class II (HLA-DP, HLA-DQ, HLA-DR) molecule restriction. Thymic RE cells also produce numerous cytokines that are important in various stages of haematopoietic cell activation and differentiation. The co-existence of pituitary hormone and neuropeptide secretion, as well as the production of a number of interleukins and growth factors, and expression of receptors for all, by RE cells is an unique molecular biological phenomenon. Thymic neuroendocrine polypeptides are the source of self antigens presented by the MHC molecules to differentiating haematopoietic stem cells. On the level of individual RE cells, the numerous projections associated with a single cell, which engulf developing lymphocytes, nurturing and guiding them in their maturation, may differ in their hormone production and/or hormone receptor expression profile, thus allowing a single cell to be involved in distinct, separate steps of the T-cell and other haematopoietic cell maturation process. Thymic RE cells represent an important cellular and humoural network within the thymic microenvironment and are involved in the homeopathic regulation mechanisms of the multicellular organism. The intrathymic T-lymphocyte selection is a complex, multistep process, influenced by several functionally specialised RE cells and under immuno-neuroendocrine regulation control reflecting the dynamic changes of the mammalian organism. PMID- 11901482 TI - Targeting apoptosis in cancer chemotherapy. AB - The aim of cancer biology is for a better understanding of the molecular basis of cancer, with the expectation that this will result in therapeutic advances and improved outcomes for patients. The discovery of apoptosis has contributed much to our understanding of the mechanisms of cell death, in both normal and neoplastic cells, and it has led to changes in the way that chemotherapy has been viewed. It is now increasingly accepted that part of the efficacy of conventional chemotherapeutic drugs is due to their ability to induce apoptosis, although this area is not without controversy. This has allowed advances in the fundamental understanding of apoptosis to have similar impacts upon cancer biology. It is now possible to construct a framework where cellular decisions about life and death can be seen as the result of a balance of pro- and anti-apoptotic signals, enacted by protein members of the Bcl-2 family, controlling mitochondrial cytochrome c release. This framework has allowed the importance of providing death signals and abrogating survival signals to both be appreciated. A range of novel approaches to the induction of apoptosis by downregulating survival signalling are described. In addition, many alternative strategies aimed at targeting particular molecular abnormalities of neoplastic cells as a means of inducing apoptosis are also under investigation and several of these are discussed. The mechanistic understanding of cell death will have profound impacts upon the practice of oncology and outlook for many patients. PMID- 11901484 TI - Meeting the market: information technology and nursing administration education. PMID- 11901483 TI - BCR-ABL as a target for novel therapeutic interventions. AB - The BCR-ABL oncogene is the result of a reciprocal translocation between the long arms of chromosome 9 and 22 t(9; 22). There is good experimental evidence demonstrating that BCR-ABL is the single causative abnormality in chronic myeloid leukaemia (CML), making it a unique model for the development of molecular targets. In addition to CML, BCR-ABL transcripts can be found in a minority of acute lymphoblastic leukaemias and very rarely in acute myeloid leukaemia (AML). Elucidating the molecular mechanisms and downstream pathways of BCR-ABL has led to the design of several novel therapeutic approaches. In this review, molecular targeting of BCR-ABL will be discussed based on the inhibition of protein tyrosine kinase activity, antisense strategies and immunomodulation. PMID- 11901485 TI - The Brain Tumor Foundation of Canada: the role of facilitators for its support groups. AB - The mission of the Brain Tumor Foundation of Canada is to fund brain tumor research, to provide patient and family support services, and to educate the public. The focus of this paper is on the role of support groups. Support groups are an important resource to support individuals with brain tumors and their families. There is an ongoing need for qualified professionals to act as facilitators for these groups. Neuroscience nurses are uniquely qualified for this role. PMID- 11901486 TI - Multiple sclerosis in childhood: understanding and caring for children with an "adult" disease. AB - Multiple sclerosis (MS) is rare in childhood but may occur more frequently than originally believed. In light of the complex nature of MS, the expanding availability of new MS treatments, and the developmental needs of pediatric patients, multidisciplinary care of pediatric patients with MS is a necessity. Our review of MS in childhood aims to increase the awareness of neuroscience nurses about MS in children and adolescents, and to expand the knowledge of pediatric nurses concerning this "adult" disease. Our experience in developing a multidisciplinary pediatric MS clinic that addresses both health and developmental needs of children with MS is presented and discussed. The nurse's role in the care of these children and their families through assessment, support, education, advocacy, referral and coordination of care is emphasized. PMID- 11901487 TI - The context, content and consequences of mothering a child with disabilities. AB - Due to recent advances in biomedical science and technology, infants and children with complex medical needs and disabling conditions are surviving and being cared for at home more than ever before. Sociocultural influences and economic policies place mothers in the home as primary caregivers of children. Their caregiving work is largely invisible. The information provided in this paper is intended to make the work of mothering a child with disabilities more visible while alerting nurses to the impact of this trend on mothers. Strategies to minimize the negative consequences are included. PMID- 11901488 TI - What to do about flat heads: preventing and treating positional occipital flattening. AB - Across Canada there has been an increasing incidence of positional occipital flattening. This increase appears to be related to the recent change in infant sleep position to supine. In this paper, two patterns of positional occipital flattening, positional plagiocephaly and positional brachycephaly, are outlined. While there is no evidence of long-term developmental or neurological problems that result from positional occipital flattening, the infant's appearance can be distressing to parents who will then seek treatment. Prevention of positional occipital flattening requires a community approach with timely screening and early intervention should the infant's skull appear flat. Treatment involves repositioning the infant coupled with physiotherapy if there is neck muscle involvement. Should repositioning alone be ineffective, a helmet or headband program may be implemented. Neuroscience nurses can work in partnership with the community to ensure prevention strategies are implemented and timely interventions initiated. PMID- 11901489 TI - Family member perspectives during acute care when a close relative is entirely dependent. PMID- 11901490 TI - Think first report--June 2000. PMID- 11901491 TI - The Mary Glover Lecture, June 14, 2000. Back to the future: reform, research, and nursing authority. PMID- 11901492 TI - Impact of a family-centred approach on a couple living with a brain tumour: a case study. AB - The diagnosis of a malignant brain tumour can be devastating for both patients and their families. Despite neurological and cancer treatment advances, there has been little progress in extending life expectancy for these patients. Cassileth et al. (1985) suggest that of all types of cancer, brain tumours cause the most psychosocial repercussions for the family. The brain tumour clinical nurse specialist (CNS) is in a strategic position to intervene with families along the continuum of care by using a family-oriented approach which can facilitate their adaptation to this health problem. The present case study will describe the illness experience of one couple where a member had a brain tumour. The process of assessment, interventions and outcomes will be highlighted. The CNS utilized the Calgary Family Assessment Model (CFAM) (Wright & Leahey, 1984) and Calgary Family Intervention Model (CFIM) (Wright & Leahey, 1994) to guide her work with this family. An example of an intervention was the illness narrative approach. In this approach the concepts of illness experience, perceptions, beliefs and support were used as part of the therapeutic modality which served to enhance the coping strategies of the couple and decreased their crisis situation. This collaborative style of nursing promoted the well-being throughout the illness trajectory of both the family and the CNS. PMID- 11901494 TI - The third millennium--a renaissance for nursing? PMID- 11901493 TI - Education and information needs identified by patients and key family members prior to surgery for a skull base neoplasm: implications for practice. AB - Many patients with extensive skull base neoplasms poorly understand the nature of their problems. Therefore, education of patients and family members is an important component of care. Research has established the benefits of pre operative education for other types of surgery and information seeking as an important method of coping. Yet in 184 references on skull base surgery (1996 1999), no case-based or research data that examined education and information needs prior to hospitalization for surgery was found. The investigators present findings from Phase 1 of a descriptive research study designed to determine the education and information needs perceived by patients and family members at their initial visit to the neurosurgeon and on admission to hospital. Data was collected, using interviews and a questionnaire, from 18 patients with skull base neoplasms and 15 key family members. The study findings provide insight into the experience of patients and families during a time period (prehospitalization) that has not been explored. Results indicate that key education needs of participants are related to the brain tumour and surgery. Findings reveal patient participants in contrast to family members had little in the way of information needs. Underlying and impacting the education and information needs is the theme of 'Hearing the News'. Relevance of the results to nursing practice in the pre operative phase is addressed. PMID- 11901495 TI - Molecular analysis of cancer. An overview. AB - Human cancers are generally characterized by acquisition of a series of somatic mutations. Molecular techniques, such as those described in this volume, have been used to identify a plethora of chromosomal translocations and mutations associated with carcinogenesis. The analysis and comparison of the array of genetic changes occurring in malignancy will enable a move toward a better understanding of cancer development. This will eventually lead to the development of improved therapies tailored to take into account the cytogenetic and molecular characteristics of specific human cancers. PMID- 11901496 TI - Use of DNA fingerprinting to detect genetic rearrangements in human cancer. PMID- 11901497 TI - Mutation analysis of large genomic regions in tumor DNA using single-strand conformation polymorphism. Lessons from the ATM gene. PMID- 11901498 TI - Mutational analysis of oncogenes and tumor suppressor genes in human cancer using denaturing gradient gel electrophoresis. PMID- 11901499 TI - Detection of mutations in human cancer using nonisotopic RNase cleavage assay. PMID- 11901500 TI - Mutational analysis of the neurofibromatosis type 1 gene in childhood myelodysplastic syndromes using a protein truncation assay. PMID- 11901501 TI - Mutation analysis of cancer using automated sequencing. PMID- 11901502 TI - Detection of differentially expressed genes in cancer using differential display. PMID- 11901503 TI - Genomewide gene expression analysis using cDNA microarrays. PMID- 11901504 TI - Gene expression profiling in cancer using cDNA microarrays. PMID- 11901505 TI - Wilms tumor gene WT1 as a tumor marker for leukemic blast cells and its role in leukemogenesis. PMID- 11901506 TI - Detection of aberrant methylation of the p15INK4B gene promoter. PMID- 11901507 TI - Clonality studies in cancer based on X chromosome inactivation phenomenon. PMID- 11901508 TI - Telomere length changes in human cancer. PMID- 11901509 TI - Measurement of telomerase activity in human hematopoietic cells and neoplastic disorders. PMID- 11901510 TI - Spectral karyotyping in cancer cytogenetics. PMID- 11901511 TI - Comparative genomic hybridization analysis. PMID- 11901512 TI - Detection of chromosomal deletions by microsatellite analysis. PMID- 11901513 TI - Detection and quantification of leukemia-specific rearrangements. PMID- 11901514 TI - Detection of chromosome abnormalities in leukemia using fluorescence in situ hybridization. PMID- 11901515 TI - Detection of t(2;5)(p23;q35) translocation by long-range PCR of genomic DNA. PMID- 11901517 TI - Due to the nursing shortage, mandatory overtime is a necessary evil. PMID- 11901518 TI - Given the nursing shortage, is mandatory overtime a necessary evil? PMID- 11901519 TI - Preparing nurse leaders. A leadership education model. PMID- 11901520 TI - Integration of research, education, and practice. When mission meets reality. PMID- 11901521 TI - The Institute for Nursing Leadership. Developing and sustaining leadership for the profession. PMID- 11901522 TI - [The health system at the hour of choice]. PMID- 11901523 TI - [One more step in zero tolerance of elder abuse]. PMID- 11901524 TI - [The first supply of milk]. PMID- 11901525 TI - [Nursing care and advanced practice]. PMID- 11901526 TI - [Creative passion: the art of being kindled]. PMID- 11901527 TI - [The future of our profession: priorities, tendencies and developments]. PMID- 11901528 TI - [Telemedicine carves out a place]. PMID- 11901529 TI - [The painful facts about scarring]. PMID- 11901531 TI - [The future of collaboration between nurses and family physicians]. PMID- 11901533 TI - [The less common road of Carolyn Joan Pepler (interview by karine Fortin)]. PMID- 11901532 TI - A profession united to define its future. PMID- 11901530 TI - ["Children are not miniature adults"]. PMID- 11901534 TI - Diabetes mellitus and the risk of prostate cancer. AB - BACKGROUND: Little is known about etiologic factors for prostate cancer. Several studies have suggested a protective effect of diabetes mellitus on the risk of prostate cancer, though a study by our group has found an elevated risk of prostate cancer following ischemic heart disease. PURPOSE: The purpose of this study was to investigate the association of diabetes mellitus with prostate cancer in the same setting in which we had found an elevated risk following ischemic heart disease. Our study differed from prior studies in utilizing a multi-racial population. Another purpose was to investigate stage-specific effects. METHODS: We conducted a hospital-based case-control study in our University Medical Center in New York City. Cases were patients with prostate cancer seen at our Medical Center between January 1, 1984 and December 31, 1986. All cases were histologically diagnosed and had undergone a biopsy or surgical procedure at Columbia-Presbyterian Medical Center (CPMC). The controls were patients who underwent a surgical procedure for benign prostatic hypertrophy (BPH) during the same time frame and were not found to have prostate cancer. Prior history of diabetes was determined by review of the medical records. Logistic regression was used to assess the association between prior history of diabetes mellitus and prior history of prostate cancer. RESULTS: We compared 320 cases to 189 controls, and found a lower risk for prostate cancer in diabetics overall (adjusted Odds Ratios (OR) 0.6, 95% Confidence Interval (CI) 0.3-1.1), though it was not statistically significant. No association was seen with Stage A prostate cancer, but there was a significant reduction in risk for stages B, C, and D combined (adjusted OR 0.47, 0.2-0.9). This effect appeared to be mainly concentrated among whites and Hispanics. CONCLUSIONS: Diabetics appear to have a lower risk of prostate cancer, though this effect may be limited to whites. An understanding of this association and its race specificity may help to explain the major difference in incidence rates for prostate cancer between blacks and whites. PMID- 11901535 TI - Immunomodulatory effects of low dose cis-Diaminedichloroplatinum (cisplatin) combined with UFT and PSK in patients with advanced colorectal cancer. AB - It is well known that cell-mediated immunity is suppressed in patients with neoplastic diseases. We have reported that soluble receptors for interleukin-2 (sIL-2R) and tumor necrosis factor (sTNF-R1) are elevated in the serum of patients with advanced colorectal cancer. The presence of these soluble receptors and immunosuppressive cytokines, including interleukin-10 (IL-10), might be important in the mechanisms of immunosuppression. cis-Diaminedichloroplatinum (cisplatin) has been reported to immunomodulate, especially when used in low dose in combination with 5-Fluorouracil (5-FU). In this study, cisplatin and UFT, a form of uracil and tegafur which is a prodrug of 5-FU, were administered with immunomodulator Polysaccharide K (PSK) to ten patients with colorectal cancer, who showed distant metastasis in the liver or lung, and the serum levels of sIL 2R and sTNF-R1 and the production of gamma-interferon (gamma-INF) and interleukin 10 by peripheral blood mononuclear cells were measured. The serum concentrations of sIL-2R and the production of IL-10 were reduced (p < 0.05) after 2 months of treatment. Thus, this combination appeared to have immunomodulative potential in patients with advanced colorectal cancer. PMID- 11901536 TI - A phase II study of topotecan in non-Hodgkin's lymphoma: an Ohio State University phase II research consortium study. AB - PURPOSE: This multicenter phase II trial evaluated the efficacy and toxicity of the topoisomerase I inhibitor topotecan (TPT, 9-dimethylaminomethyl-10 hydroxycamptothecin) in patients with refractory or relapsing non-Hodgkin's lymphoma. PATIENTS AND METHODS: Thirty-two patients with previously treated non Hodgkin's lymphoma were accrued in this study from June 1992 to June 1997. Patients were eligible if they had measurable disease and had received two or less chemotherapy treatments for indolent lymphoma or no more than one treatment for aggressive lymphoma. Nineteen patients with aggressive lymphoma including seven with transformed indolent disease and 11 patients with indolent disease were treated. Two additional patients had both indolent and aggressive lymphoma in the bone marrow and lymph nodes at entrance into the study. Topotecan was administered as a 30-min infusion at a dose of 1.25 mg/m2 for five days and repeated at three week intervals. RESULTS: Thirty-two patients were evaluable for toxicity and 29 patients were evaluable for response. There were five objective responses including two complete remissions and three partial remissions (17%; CI 4-30%). Four of the five responders had aggressive disease and had been responsive to prior chemotherapy. The duration of remissions were short, lasting a median of 2 months (range 2-52 months). The major toxicity seen was myelosuppression with 28 of the 32 patients having grade three or greater granulocytopenia. CONCLUSION: Topotecan given as a five day 30 min infusion at a dose of 1.25 mg/m2 has modest activity in previously treated non-Hodgkin's lymphoma as compared to other active agents. PMID- 11901537 TI - Toxicity of a 24-hour infusion of gemcitabine in biliary tract and pancreatic cancer: a pilot study. AB - The antitumor activity of gemcitabine is not dose-response related but schedule dependent. Based on the results of a published phase I study in patients with nonsmall-cell lung cancer we started a pilot study of a 24-hr infusion of gemcitabine in patients with adenocarcinoma of the pancreas and biliary tract cancer. Twenty-five patients were enrolled and received a 24-hr infusion of gemcitabine once weekly on three consecutive out of 4 weeks. Dose levels of gemcitabine ranged from 100 to 150 mg/m2. One of 13 chemotherapy-naive patients had a partial response. Dose-limiting toxicity (DLT) was thrombocytopenia in pretreated patients and neutropenia in chemotherapy-naive patients. Other toxicities were oral mucositis, fever, flu-like symptoms, and asthenia. Maximum tolerated dose (MTD), especially in pretreated patients, was 100 mg/m2. PMID- 11901538 TI - A phase II trial of interferon-alpha and toremifene in advanced renal cell cancer patients. AB - Treatment options for patients with metastatic renal cell carcinoma are limited. Interferon-alpha has an overall response rate of 10-15% in phase II and III clinical trials and is considered a standard option for patients. Though the anti estrogen toremifene has shown only modest single agent activity in renal cell carcinoma, evidence for synergy of anti-estrogens with interferon-alpha exists in renal cell and other cancers. Therefore, a phase II trial was undertaken to test the combination of interferon-alpha and toremifene in advanced renal cell carcinoma. Thirteen patients with measurable metastatic or unresectable local disease were treated with interferon-alpha at a dose of 5 million units/m2 three times a week and daily oral toremifene at 300 mg daily in divided doses. Patients were treated for 12 weeks and then restaged. Clinical response was the primary endpoint of the trial. Four patients (31%) had evidence of stable disease at 12 weeks, while the remaining nine patients (69%) progressed on treatment. Toxicity was moderate, with grade 2 or 3 fatigue, nausea and anorexia each noted in 31% of patients. We conclude that the combination of interferon-alpha plus toremifene demonstrates no significant activity in advanced renal cell carcinoma. PMID- 11901540 TI - Comparison of the expression of P-glycoprotein, Ki-67, and P-53 to technetium-99m tetrofosmin mammoscintigraphic findings. AB - Adjuvant and neoadjuvant chemotherapy is being used increasingly in the treatment of breast cancer patients. However, drug resistance plays an important role in the failure of chemotherapy in breast cancer. The aim of this study was to compare technetium-99m tetrofosmin (Tc-TETRO) mammoscintigraphic findings with the expression of drug resistance proteins (p-glycoprotein [Pgp], Ki-67 and mutant p53) in human breast cancer tissues. Thirty patients diagnosed with infiltrating ductal breast cancer underwent Tc-TETRO mammoscintigraphy before surgery or biopsy. Protein expression was investigated by immunohistochemical analyses on multiple nonconsecutive sections from surgery or biopsy specimens. Tumor to background (T/B) ratios calculated by Tc-TETRO mammoscintigraphic findings were correlated with protein expression determined by immunohistochemical analyses. The Tc-TETRO T/B ratios were significantly lower for tumors in 12 patients with positive Pgp expression than for those in 18 patients with negative expression (1.19 +/- 0.13 and 1.94 +/- 0.33, p value < 0.01). However, differences in Tc-TETRO T/B ratios between the patients with positive and negative Ki-67 or mutant p53 expression were not found in this study. Our data confirmed that Tc-TETRO mammoscintigraphic findings are useful for determination of the presence of Pgp expression in breast cancer patients, but no significant relations between Tc-TETRO mammoscintigraphic findings and Ki67 or mutant p53 were found. PMID- 11901539 TI - A phase II study of CI-980 in previously untreated extensive small cell lung cancer: an Ohio State University phase II research consortium study. AB - CI-980, (ethyl (S)-(5-amino-1,2-dihydro-2-methyl-3-phenylpyrido[3,4-b] pyrazine-7 yl) carbamate 2-hydroxyethansulfonate (1:1)), is a water-soluble mitotic inhibitor. It acts by binding to the colchicine-binding site on tubulin, a site different from that of the vinca alkaloids, inhibiting tubulin polymerization. Cells exposed to CI-980 accumulate in M phase and die. In preclinical tumor models, CI-980 showed a broad spectrum of activity, including in multi-drug resistant tumor cell lines, with activity at least equal to that of vincristine. Extensive small cell lung cancer, despite its responsiveness to chemotherapy, is usually an incurable disease with survival in patients of less than one year. Due to the preclinical activity of CI-980 and its similar mechanism of action to drugs effective in small cell lung cancer, a phase II trial in extensive small cell lung cancer was initiated by The Ohio State University Phase II Research Consortium. A "window of opportunity" design was chosen where a short six-week trial of the drug was given unless there was significant objective response. Twelve patients were entered in the study and underwent a total of 16 cycles of chemotherapy. The median age of the patients was 54 years old (range 34-71) and performance status was ECOG 0 (four patients), ECOG 1 (seven patients), and ECOG 2 (one patient). The patients were treated with a 72-hr infusion at a dose of 4.5 mg/m2/day. Toxicity was predominantly myelosuppression with granulocytopenia (nine episodes), and anemia (seven episodes). There were no objective responses with 11 patients being removed from study due to progressive disease. Evaluation of leukocyte microtubule structure in peripheral blood revealed microtubule depolymerization, which was seen after treatment (t = 72 hr) and was reversible within 24 hr of stopping the drug. We conclude that despite antitumor activity demonstrated in preclinical studies, CI-980 does not have biological activity in previously untreated small cell lung cancer at this dose and infusion protocol. PMID- 11901541 TI - Successful treatment of transitional cell carcinoma of the urethra with chemotherapy. PMID- 11901542 TI - Peptide vaccines for cancer. AB - The revelation that immune cytolytic and helper T-cells recognize intracellularly degraded peptides processed via the proteosome apparatus, inserted into the endoplasmic reticulum and transported to the surface for association with major histocompatibility locus (MHC) molecules on specialized antigen-presenting cells has revolutionized the cancer vaccine field. Understanding the science of antigen processing and presentation has provided new reagents, delivery systems, and new investigative leads for the generation of immune responses against cancer. The cloning of tumor antigen genes has proceeded rapidly in melanoma, due to the ease with which melanoma-specific T-cells can be propagated in vitro, but breast, cervix, and lung cancer are not far behind. The cloning and identification of tumor antigens recognized by T-cells and data from initial clinical trials with peptides vaccines derived from those antigens are presented. PMID- 11901543 TI - Cancer/testis antigens: structural and immunobiological properties. AB - Characterization of tumor-associated antigens recognized by cytotoxic T lymphocytes which has evolved during recent years opens new possibilities for specific anti-cancer immunotherapy. Among different groups of tumor-associated antigens, cancer/testis (CT) antigens (expressed in many tumors and among normal tissues only in testes) represent the most perspective antigens for immunotherapy because of their broad tumor-specific expression. More than 50 CT antigens have been described so far and, for many of them, epitopes recognized by T lymphocytes have been identified. The most studied group of CT antigens is the MAGE proteins, which form the so-called MAGE superfamily, together with some MAGE-like proteins that have a different distribution than classical CT antigens. The MAGE superfamily includes five families: MAGE-A, MAGE-B, MAGE-C, MAGE-D, and necdin. Comparison of the structure of members of MAGE superfamily points to the existence of a domain organization of these proteins. The central, core domain (second domain) is highly conservative. The first domain is homologous among MAGE family members with a CT expression, but unique for each member of the MAGE-D and necdin families. In addition to the homology of the central domain, the third domain is also homologous among all members of MAGE superfamily, but to a much lesser extent. The MAGE-D proteins contain an additional, fourth domain, which in the case of MAGE-D3 coincides with trophinin, a separate molecule described previously as an adhesion molecule that participates in embryo implantation. The structural classification of the members of MAGE superfamily might help in the future to understand the biological function of MAGE proteins. One important property of the CT antigens is the up-regulation of their expression by DNA demethylating agents, indicating a possible mechanism for their re-expression in tumors. One of the implications of this particular property could be that a combination of immunotherapy targeting CT antigens with chemotherapy inducing up regulation of CT antigens might result in more efficient tumor eradication. PMID- 11901544 TI - New opportunities in chemoprevention research. AB - The importance of prevention in reducing the morbidity and mortality from cancer has been widely recognized. With the demonstration of tamoxifen's ability to prevent breast cancer in women, the feasibility of cancer chemoprevention in humans is now established. Future clinical chemoprevention studies should focus on phytochemicals, cancer preventive compounds in fruits, vegetables and other plants. Many phytochemicals are excellent potential chemopreventive agents, because, in addition to their cancer preventive effects, they are relatively non toxic and inexpensive, they can be taken orally and some of them have other health benefits as well. New opportunities in clinical chemoprevention research include investigating chemopreventive effects of phytochemicals and conducting studies in patients with cancer. There is also a great need to investigate potential benefits and risks of administering phytochemicals before, during or after conventional therapies, such as surgery, chemotherapy, radiation or hormonal therapy. In addition, administration of chemopreventive agents prior to surgery provides an opportunity to investigate the modulation of genetic and epigenetic pathways by putative cancer preventive compounds and nutrients. PMID- 11901545 TI - Oxaliplatin: a review of evolving concepts. PMID- 11901546 TI - Malignant worms: what cancer research can learn from C. elegans. AB - Developmental processes in the nematode C. elegans are controlled by pathways of gene functions that are analogous to those used in mammals. Hence, genetic studies in C. elegans have helped build the frameworks for these regulatory pathways. Many homologs of human genes that are targets for mutation in cancer have been found to function at distinct steps within such genetic pathways. This way, studies in C. elegans have provided important clues about the functions of human oncogenes and tumor suppressors. Understanding how human cancer genes function and act in signaling cascades is of great importance. This information reveals what kind of molecular changes contribute to the process of cell transformation. Moreover, additional candidate oncogenes and tumor suppressors may be revealed by identifying the functional partners of genes with an established role in cancer. Furthermore, identifying a cascade of gene functions increases the number of potential targets for therapeutic intervention, as blocking either one of multiple genes may interfere with signal transduction through the pathway. Simultaneous approaches in a number of different model systems act synergistically in solving pathways of gene functions. By using multiple models, the field takes advantage of the strengths of each system and circumvents its limitations. As one of the most powerful genetic animal systems, C. elegans will continue to reveal new mammalian signaling components. In addition, now that the C. elegans genome sequence has been completed, an increasing number of researchers are likely to discover homologs of human disease genes in the nematode and to analyze gene function in the worm model. Combined with the great potential of this animal in drug screens, it is simple to predict that C. elegans will worm its way deeper and deeper into cancer research. PMID- 11901547 TI - Melatonin protects against oxidative stress caused by delta-aminolevulinic acid: implications for cancer reduction. AB - delta-Aminolevulinic acid (ALA) is a precursor of haem. The increased concentration of ALA is typically related to acute intermittent porphyria, hereditary tyrosinemia, and lead poisoning. delta-Aminolevulinic acid produced in excess accumulates in a number of organs, causes oxidative damage, and often leads to cancer. Melatonin (N-acetyl-5-methoxytryptamine) is a well-known antioxidant, free radical scavenger, and exhibits anticarcinogenic properties. It protects DNA, lipids, and proteins from oxidative damage. The protective effects of melatonin against ALA-induced oxidation of guanine bases, lipid peroxidation, and alterations in membrane fluidity in several organs have been documented. There is an inverse relationship between melatonin and ALA concentrations in both experimental and clinical conditions of porphyria. The marked efficacy of melatonin in protecting against ALA-related oxidative stress, its oncostatic properties, and low toxicity constitute reasons to consider the use of this indoleamine as a co-treatment in patients suffering from disturbances related to ALA accumulation. PMID- 11901548 TI - Oxaliplatin: new opportunities for the treatment of gastrointestinal cancer. PMID- 11901549 TI - Colorectal cancer screening under the age of 50 is less cost-effective. PMID- 11901550 TI - On the frame of reference in psychotherapy and psychoanalysis. PMID- 11901551 TI - Psychoanalysis and creativity: beyond Freud and Waelder. PMID- 11901552 TI - Cluster B personality traits and attachment. PMID- 11901553 TI - Countertransference factors in the psychology of psychopharmacology. PMID- 11901554 TI - Angels in the architecture: contemporary case of a orphic functioning. PMID- 11901555 TI - Working through in psychoanalytic psychotherapy: an alternative and complementary path. PMID- 11901557 TI - Fromm's concern with feminine values. PMID- 11901556 TI - Toward a neurobiology of the unconscious. PMID- 11901558 TI - Of time, narrative, and cast away. PMID- 11901559 TI - Narcissism in collecting art and antiques. PMID- 11901560 TI - The currency of mythology. PMID- 11901561 TI - The contemporary failure of nerve and the crisis in psychoanalysis. AB - The American Academy of Psychoanalysis is undergoing an identity crisis at this time, which is at least to a large extent a function of the whole current identity crisis in the field of psychoanalysis itself. In order to better understand this crisis, in this article I have first reviewed a similar situation which occurred in the history of classical Greece. Plato's famous Academy underwent a progressive deterioration and disintegration and fragmentation, until it ended up merely the handmaiden of another discipline, Christian theology, for a thousand years. I then propose that the identity crisis in psychoanalysis today has to do with our failure of nerve in the teeth of the abusive behavior of insurance companies regarding the payment for psychoanalysis and the current cultural ambience demanding "fast-fast-fast" relief. I call in this article for a return to Freud's basic principles as a focus for our identity. Of course we cannot ignore new discoveries in neurobiology if they are well established, or what we learn from the study of enactments in the here-and-how of the analytic procedure. Certainly the findings of Freud that are contradicted by firmly accepted empirical findings in neurobiology and other disciplines call for revision of some of his ideas, as do his mistaken views on the psychology of women and on certain other topics such as art, religion, and evolutionary biology. But this should not be permitted to blur our continuing focus on the fundamental principles of the clinical practice of psychoanalysis as Freud developed them over his lifetime. In this article I briefly reviewed those basic principles and proposed that we employ them as the basis for our identity as psychoanalysts and psychoanalytic psychiatrists. It represents a failure of nerve to drift this way and that with current fads and with the continuously deteriorating ambiance of our culture as the world slides into rampant global capitalism. Franz Alexander said years ago that psychoanalytic psychotherapy is one of the last remnants of the humanistic ideal, focussing on the individual unique person and his or her transcendent possibilities as well as maladaptive pathology. This article represents a clarion call for a debate on the identity of the American Academy of Psychoanalysis and what it stands for, which can only be clarified if we have a sharp focus on what we basically mean by "psychoanalysis." As Saul Bellow puts it (Atlas, 2000) in discussing the disappointing current situation for the arts and the humanistic disciplines, the intelligent public is waiting to hear from these disciplines what it cannot hear from pure science: Out of the struggle at the center has come an immense, painful longing for a broader, more flexible, fuller, more coherent, more comprehensive account of what we human beings are, who we are, and what this life is for...the individual struggles with dehumanization for the possession of his soul, (p.462). Below points out, in talking about writers, and in a discussion equally applicable to psychoanalysts, that if we do not "come again into the center it will not be because the center is preempted. It is not. [We] are free to enter if [we] so wish" (p.462). PMID- 11901562 TI - Force health protection: 10 years of lessons learned by the Department of Defense. AB - The Department of Defense has applied lessons learned since the Persian Gulf War to develop the force health protection (FHP) strategy. The goal of this new, unified strategy is to protect the health of military members from medical and environmental hazards associated with military service to the maximum extent possible. FHP is an evolving strategy that seeks to balance the military health system's responsibilities to promote and sustain health and wellness throughout each person's military service; prevent acute and chronic illnesses and injuries during training and deployment; and rapidly stabilize, treat, and evacuate casualties. In addition, FHP demands a continuous assessment of the current and future health of military members through medical surveillance, longitudinal health studies, adequate medical record documentation, and clinical follow-up. Effective communication with military members, leaders, veterans, families, and the public regarding military members' health status and the health risks of military service is a key element of the FHP strategy. PMID- 11901563 TI - The Institute of Medicine's independent scientific assessment of Gulf War health issues. AB - The Institute of Medicine has frequently been the source of expert advice to the government and others on questions related to health and medicine. Such has been the case as Congress, federal agencies, and veterans attempt to resolve conflicts and develop policies to address the health concerns of Persian Gulf War veterans. Twelve reports issued by Institute of Medicine committees address what is known about exposures and illnesses in Gulf War veterans and what additional information is needed, how clinical programs for Gulf War veterans could be improved, and what strategies could help prevent or better address similar health problems in the future. The Institute of Medicine reports recommend guidelines and interventions to treat sufferers of medically unexplained symptoms, longitudinal studies to measure changes in health status, and improved risk communication. They emphasize the need for the maintenance of retrievable electronic records of baseline health status, of exposures, and of health events that occur during a service member's career. PMID- 11901564 TI - Innovation in veterans' health care and assistance: the Department of Veterans Affairs 10 years after the Gulf War. AB - The Department of Veterans Affairs (VA) has responded to significant challenges in treating and compensating Persian Gulf War veterans by adapting existing programs and developing new ones. The VA established a Gulf War health examination registry and expanded existing "Vet Centers" to provide assistance to Gulf War veterans. Health care eligibility income limitations were eliminated. Outreach efforts included a national newsletter, veterans' organization briefings, and other products. The VA is developing targeted training programs and continuing medical education for health care providers. Numerous major research initiatives have begun. Innovations include the establishment of environmental hazards research centers, clinical demonstration projects, and centers for the study of war-related illness. These efforts required increased coordination among federal agencies and collaboration with other countries. In a precedent-setting development, Congress gave the VA authority to compensate certain veterans with undiagnosed illnesses. Veterans from future conflicts and peacekeeping missions can expect improved services from the VA as a result of these initiatives. PMID- 11901565 TI - Oxygen supplementation of the Impact 754 ventilator. AB - Oxygen supplementation of available transport ventilators may become a relevant issue in environments in which pressurized oxygen is not available. Ventilators available for use by U.S. Army personnel in austere locations were compared. The results clearly demonstrate the effectiveness of off-the-shelf oxygen generators providing adequate oxygen supplementation with both portable ventilators. Others should be able to use this information for the benefit of their patients in field environments, especially forward surgical teams and those in austere health care locations. PMID- 11901566 TI - Physician-to-physician consultation via electronic mail: the Walter Reed Army Medical Center Ask a Doc system. AB - INTRODUCTION: Physician-to-physician consultation and discussion have traditionally been conducted by telephone, paper, and "curbside" (face to face meetings). The implementation and use of physician-to-physician consultation via electronic mail in a military health care system has not been reported previously. METHODS: The group mail function of the Composite Health Care System, the main outpatient medical automation system for the Department of Defense, was modified to create mailgroups for every specialty of the Walter Reed Army Medical Center to facilitate ease of physician-to-physician consultation. This modification was called the "Ask a Doc" system. The system was deployed to a 21 state health care network among triservice participants. RESULTS: There were 3,121 consultations logged from April 22, 1998, to December 31, 2000. Growth in use expanded initially and was sustained during a 3-year period. Average response time to consultations was less than 1 day (11.93 hours). Additional training and maintenance requirements were minimal. In general, the use of electronic consultation mirrored that of clinical practice. Most specialty consultations involved the disciplines of internal medicine. CONCLUSIONS: Use of the Ask a Doc system was representative of total clinical workload and increased access to specialty medical care over a wide geographic area. The distribution of use indicated that user statistics were legitimate, and quality improvement programs could easily troubleshoot the system. Ask a Doc was inserted into a regional health care network with minimal cost to support and implement and was sustained with very little effort for 3 years. Barriers to even wider use currently include lack of secure communications and the difficulty in assigning workload credit for electronic consultations. PMID- 11901567 TI - Ambulatory medical visits among anthrax-vaccinated and unvaccinated personnel after return from southwest Asia. AB - The Department of Defense launched a mandatory anthrax immunization program for military personnel in December 1997. This program has been criticized for many reasons, including concern over side effects. This study was designed to give a quick answer to the question of whether vaccinated persons who deployed to southwest Asia were more likely to seek medical care upon their return than their unvaccinated counterparts. The results demonstrated that there was no greater risk for vaccinated persons to have a diagnosis recorded in the Ambulatory Data System (0.96 RR) than unvaccinated persons. In addition, there was no significant increased risk for a recorded diagnosis in any 1 of the 17 International Classification of Diseases, Ninth Revision, categories or for 16 specific adverse health conditions. PMID- 11901568 TI - Does the diagnosis of coronary calcification with electron beam computed tomography motivate behavioral change in smokers? AB - BACKGROUND: Electron beam computed tomography is promoted as a test that may enhance patient motivation to alter cardiovascular risk behaviors. METHODS: We surveyed a consecutive sample of asymptomatic, active smokers (N = 144) who underwent screening electron beam computed tomography about their current motivation to alter their smoking behavior in the context of their tomography results. RESULTS: Patients with coronary artery calcification (42% of the sample) were more likely to perceive increased cardiovascular risk (42 vs. 13%; p < 0.01). Overall, most patients (59%) rated themselves as more motivated to quit smoking after electron beam computed tomography, but there was no relationship between motivational levels or smoking behavioral change and the presence of coronary artery calcification. CONCLUSIONS: In smokers predominantly self referred for electron beam computed tomography, the presence of coronary artery calcification does not appear to influence motivation for smoking cessation or smoking behavior. PMID- 11901569 TI - Pertussis in a military and military beneficiary population: case series and review of the literature. AB - Three cases of pertussis (whooping cough) identified in a military emergency department are reported. Two of these cases involved infants with typical presentations. One of these infants was too young to have received immunizations, and the other child was only partially immunized. The third case involved an active duty soldier with a chronic cough. Pertussis has become increasingly important as a cause of chronic cough in adults. As a result of the infectivity of this organism, close-quarter situations, such as day care centers and military barracks, create the opportunity for substantial person-to-person transmission. Typical and atypical clinical presentations of pertussis are discussed, with an emphasis on currently available diagnostic modalities. The epidemiology and pathophysiology of this disease are also reviewed. The medical management of active duty soldiers and their dependents (both pediatric and adult) with this largely underappreciated infection and their close contacts is presented. PMID- 11901570 TI - An epidemiological survey of Clostridium perfringens-associated enterotoxemia at an army veterinary treatment facility. AB - The purpose of this study was to determine the incidence of Clostridium perfringens-associated vomiting and diarrhea in canine and feline patients at the Okinawa Branch Veterinary Service and to establish a relationship between dietary indiscretion and the presence of clostridial enterotoxemia. Stool samples were obtained from 38 control animals and 44 animals with vomiting and diarrhea during a 3-month period. For each sample, fecal cytology were examined for the presence of C. perfringens, white blood cells, and red blood cells. A survey was also completed by the animal's owner, which provided the researcher with information on the owner's pet feeding practices. After the data were analyzed, 27% of animals with vomiting and diarrhea were found to have C. perfringens upon examination of their fecal samples (p < 0.05), compared with 11% for control animals. Results of our survey revealed that when owners were questioned regarding their feeding practices, 45% of the animals with C. perfringens positive fecal samples and with clinical signs of vomiting and/or diarrhea had an episode of dietary indiscretion soon before the illness (p < 0.05). Therefore, we conclude that C. perfringens is an important cause of vomiting and diarrhea in our patient population and that a causal relationship may exist between dietary indiscretion and C. perfringens-associated intestinal disease. PMID- 11901571 TI - Determination of the reasons for unfitness for military service in Turkey. AB - This study aims to evaluate the frequencies and diagnosis types of cases in which young adult males were classified as unfit for military service before or during military service. This cross-sectional study was conducted between November 1998 and October 1999 by investigating the military medical board records of 21,985 "unfit" cases. Unfit for military service decisions were based on any of 436 different diagnoses. Twenty-three diagnosis types, each with a frequency of greater than 1.0%, collectively accounted for the majority of cases (59.2%). Initial medical examinations before service detected 64.0% of cases, whereas the remaining 36.0% were detected during service. Initial medical examinations can be a remarkable data source to determine the health profile of a young adult male population. High rates of failure in initial medical examinations call for improvements to initial medical examination procedures, which should eventually lead to cost savings for the military. PMID- 11901572 TI - Accuracy of prescription medication reporting among active duty U.S. Army dental patients. AB - Dentist routinely ask patients about medication usage before dental treatment. The purpose of this study was to determine if active duty personnel relate accurate information about medications they have been prescribed in the 6 months before an annual dental examination. This study also examined whether patients knew the dosage of the medication they were prescribed. The information in this study was obtained from 100 active duty soldiers at their dental examinations. The data were verified through the Composite Health Care System computer information system. In this study, 35% of patients had been prescribed at least one medication. Fifty-one percent of the patients prescribed medication in this study accurately reported their prescription history in the 6 months before their dental examinations. Forty-five percent of the patients that reported being prescribed medication actually knew the correct dosage of the medication they were prescribed. PMID- 11901573 TI - Case management of asthma for family practice patients: a pilot study. AB - OBJECTIVE: Asthma is a chronic inflammatory disease of the airways that affects between 14 and 15 million persons in the United States. It is responsible for more than 470,000 hospitalizations annually and an estimated $6 billion in total medical cost. The Asthma Case Management Program instituted at our hospital is based on the concept of patient self-management. It involves patient education, a home treatment plan (HTP), and physician/nurse follow-up. This study was performed to determine whether an organized case management program improves outcomes and cost in family practice asthma patients. STUDY DESIGN: A prospective cohort study of 48 asthmatic patients was conducted with a retrospective review. Data were obtained from health, pharmacy, and computer records. PATIENTS AND METHODS: Forty-eight asthma patients, ages 1 year to adult, assigned to the Family Practice Department were enrolled in the Asthma Case Management Program. This cohort was offered asthma education from a provider trained in national asthma guidelines. Most of these patients received a coordinated HTP completed by their primary care provider. The asthma case manager initiated regularly scheduled nursing follow-up. Hospital admissions, emergency department and clinic visits, number of chest radiographs, and use of beta 2 agonists and anti inflammatory drugs were recorded for a mean of 6 months before and 6 months after the intervention. Twenty-eight patients who had received the HTP as part of their intervention were compared with 12 patients who did not. A cost analysis was completed. RESULTS: All measured parameters showed favorable changes after intervention. Statistically significant decreases in clinic visits, chest radiographs ordered, beta 2 agonists, and oral anti-inflammatory drugs were obtained with the 28 patients who received the HTP. Six-month resource savings after intervention were estimated at $19,677.42 ($491.90 per patient). Ninety three percent of these savings are attributed to those patients with the HTP. There were no statistically significant improvements and considerably fewer savings for those patients not on the HTP. CONCLUSION: A combined intervention consisting of patient education, a coordinated self-monitoring plan, and patient follow-up was associated with improved care and economic outcomes in this group. The greatest clinical improvement and resource savings are clearly seen in those patients who have received the HTP as part of their asthma case management. Every effort should be made to include the HTP as the central part of asthma case management. PMID- 11901575 TI - Effectiveness in meeting recommended standards for annual diabetic eye examinations at a veterans health administration facility. AB - This study determined the effectiveness of one Veterans Affairs Medical Center facility at providing annual diabetic eye examinations. A medical records review of a simple random sample of 350 diabetic patients was conducted to assess the impact of delayed access to care, practitioner referral patterns, patient no-show rates, and sample sizes used in performance reports. A 55.6% adherence rate was found, which increased to 81% when adjusted for future scheduled appointments. A high rate of physician referral for diabetic eye care was documented (68.9%), with a 10.2% patient no-show rate. The primary factor adversely affecting diabetic eye examination rates at this facility was a delayed access to care, complicated by an increased demand for eye care services. Also, sample sizes used in official performance reports were not sufficient to accurately determine true examination rates at the local level. PMID- 11901574 TI - Racial variations in Department of Veterans Affairs ambulatory care use and unmet health care needs. AB - Our objective was to describe racial/ethnic variations in Department of Veterans Affairs (VA) ambulatory care use and its association with the presence of unmet health care needs. Using the 1992 National Survey of Veterans, we examined race/ethnicity and unmet health care need for ambulatory care users of VA and non VA facilities. Black and Hispanic veterans were more likely to report any VA use. In unadjusted analyses, American Indian/Eskimo, Hispanic, and black veterans were 4.4, 2.5, and 1.9 times more likely, respectively, than white veterans to report an inability to get needed care. Adjusting for VA ambulatory care use diminished the disparity in inability to get needed care between American Indian/Eskimo or Hispanic veterans and white veterans and eliminated the disparity between black and white veterans. Our findings support the VA's role as a medical safety net provider and suggest that VA ambulatory care use is effective in mitigating health-related racial disparities for some veterans. Additional facilitators for reducing unmet need should be explored. PMID- 11901576 TI - Changes in food intake and body weight associated with basic combat training. AB - This research project evaluated changes in food selections, food intake, and body weight during 8 weeks of basic combat training (BCT). During the first week of BCT, 139 soldiers from two companies volunteered for participation in the study. In the eighth week of BCT, 92 soldiers were available for retesting. A digital photography method for measuring food selections and food intake was developed for this study. Fruit intake of soldiers was very low at the beginning and end of BCT. Food intake for grains and milk products was low during the first week of BCT but improved by the end of BCT. Average body weight decreased during the 8 weeks of BCT, but heavier soldiers tended to lose weight and thinner soldiers tended to gain weight. These findings suggest that the overall effect of BCT was a trend toward improvement of healthy eating and healthy body weight. PMID- 11901577 TI - Effectiveness of two versions of a sexually transmitted diseases/human immunodeficiency virus prevention program. AB - Little is known about the comparative effectiveness of human immunodeficiency virus prevention interventions that differ in duration but contain similar content. The objective of this study was to evaluate and compare the effectiveness of two versions (6 hours vs. 3 hours) of a behavioral intervention called the STD/HIV Intervention Program (SHIP) in a sample of Marines. Marines were exposed to either a 6-hour or a 3-hour version of SHIP. Comparisons of pre test and post-test knowledge, attitude, and behavioral intention scores revealed similar results for both versions. For both versions of the intervention, scores on sexually transmitted diseases/human immunodeficiency virus knowledge were significantly higher after the intervention. Both the 6-hour and the 3-hour versions of SHIP also led to significant increases on scales measuring social norms and behavioral intentions. The two versions of SHIP appeared to be of comparable effectiveness for producing short-term changes in knowledge, attitudes, and behavioral intentions. PMID- 11901578 TI - Training military surgeons: a challenge for the future. AB - The last 10 years has seen a reduction in defense spending and a contraction in military force size in all NATO countries. This has had a direct effect on military medical capability. In some allied countries, this reduction has extended to the virtual disappearance of independent military hospitals. Military surgeons are now few in number, and fewer still have had recent operational experience. This article addresses the problem and offers some solutions. PMID- 11901579 TI - Recurrent syncope in a 31-year-old army staff sergeant. PMID- 11901580 TI - Biocompatibility of electrochemically activated aqueous solutions: an animal study. AB - The purpose of the study was to evaluate the biocompatibility of electrochemically activated aqueous solutions on experimental animals. Nine rabbits were subjected to acute eye irritation/corrosion tests, nine rabbits to acute dermal irritation/corrosion and 30 rats to acute oral toxicity (LD50) tests. No significant negative effects were noted. Considering the relatively high levels of exposure of the animals to the solutions and the low levels in the anticipated real clinical situation it is argued that these solutions are indeed biocompatible. PMID- 11901581 TI - The influence of orthodontic bracket base diameter and mesh size on bond strength. AB - Previous studies have shown that the size of the foil mesh and surface area of the bracket base correlated to bond strength to tooth enamel. The purpose of this study was to investigate the in vitro shear bond strength (SBS) of orthodontic brackets with 80 and 100 gauge mesh bases as well as mini and standard size bases. Eighty extracted human premolar teeth were randomly allocated into four groups of twenty teeth each. Premolar brackets of different mesh and bracket base area sizes were bonded to the enamel with a conventional 'two paste' orthodontic bonding agent. The SBS was determined and the bond failure sites were assessed using light-optical microscope and SEM. Bond failure occurred in all groups primarily at the bracket/adhesive interface. Mean values for SBS were 9.97 +/- 2.94 MPa and 10.72 +/- 2.54 MPa for 80 gauge mini and standard size respectively, and 10.45 +/- 3.27 MPa and 11.39 +/- 3.32 MPa for 100 gauge mini and standard size. A one-way ANOVA and an unpaired t-test revealed no significant difference in mean SBS (P > 0.05) between 80 and 100 gauge mini and standard size brackets, and no significant difference in mean SBS between brackets of the same surface area with a different gauge mesh size. The clinical relevance of this finding is that the clinician can select smaller brackets with no reduction in effectiveness of the treatment procedure. PMID- 11901582 TI - The role of the dentist in the prevention and early diagnosis of oral cancer. PMID- 11901583 TI - [Lingualized occlusion in the South African context]. AB - The search for the ideal artificial tooth arrangement that maximizes denture stability, comfort, aesthetics, and function has occupied the dental literature for many years and still continues to do so. Of the many occlusal schemes that have been presented to the dental profession, that of lingualized occlusion has emerged as one of the more popular. The popularity of lingualized occlusion stems from the simplicity and flexibility of the concept and from its wide application to clinical practice (Parr & Ivanhoe, 1996). The registration of a repeatable correct centric jaw relation is not always possible. We don't know whether the patient will use centric relation during normal function. It is therefore useful to provide the patient with some freedom of movement around centric. lingualized occlusion provides freedom in centric. For many dentists the arrangement of artificial denture teeth into balanced occlusion is difficult and time consuming. As a result this task is most often performed by the dental technician. In the South African countryside dental laboratories are often far away. If dentists perform the arrangement of the denture teeth, time and costs can be saved. The mounting of denture teeth in lingualized occlusion is simple and fast. This will motivate dentists to arrange denture teeth themselves, with obvious benefits for both the patient and the dentist. The School of Oral Health Sciences of the University of Stellenbosch teaches this concept to its undergraduate students in order to improve the prosthetic service to the large edentulous population of South Africa. PMID- 11901584 TI - Use of technopolymer clasps in prostheses for patients due to have radiation therapy. AB - Patients with large oral defects as a result of a maxillectomy procedure, who are due to have postoperative radiotherapy, need to have the density of the defect restored to ensure standardised radiation distribution. This is achieved with various types of boluses that often require tissue surface positioning stents to help support them. Traditional metal-clasp retained stents were discarded as the clasps caused backscatter of the radiation beams. A radiolucent material was needed to retain these prostheses. Dental D, an acetal resin was used in the fabrication of a positioning stent. It was assessed in terms of ease of manufacture, cost, fit, retention and radiolucency. The material was found to be more costly and time-consuming to manufacture than conventional metal-retained acrylic resin prostheses, but its radiolucency made it ideal for use in patients during radiotherapy treatment. Future studies will determine if the long-term strength of the material will allow the clasps to be used in a definitive prosthesis and thus help offset the initial time and costs. PMID- 11901585 TI - The context of a dental school in Africa. PMID- 11901586 TI - Iatrogenic oral ulceration. PMID- 11901587 TI - Feline visual changes associated with enrofloxacin. PMID- 11901588 TI - An ethicist's commentary on veterinarians producing autogenous vaccines and compounding antimicrobial drugs. PMID- 11901589 TI - Veterinary schools could lose accreditation. PMID- 11901590 TI - Veterinarians and SPCAs: an essential partnership. PMID- 11901591 TI - An alternative to undergraduate streaming and limited licensure. PMID- 11901592 TI - Clinical, radiographic, endoscopic, bronchoalveolar lavage and lung biopsy findings in horses with exercise-induced pulmonary hemorrhage. AB - A clinical study was performed to determine whether clinical, endoscopic, radiographic, bronchoalveolar lavage (BAL) cytological, and pulmonary biopsy findings could be correlated in horses with exercise-induced pulmonary hemorrhage (EIPH) compared with controls. Racing standardbred horses were selected as either EIPH (n = 10) or control (n = 10), based on repeated postexertional endoscopy of the lower airways. Complete physical and respiratory examinations were performed and blood samples were submitted for arterial blood gas analysis, hematologic study, and fibrinogen determination. Bilateral chest radiographs were taken with the horse standing, and a BAL sample was obtained for cytological examination. Lung was biopsied transcutaneously. Weighted scores were calculated for clinical, radiographic, and pulmonary biopsy findings. The conclusion was that only routine physical examination may help the clinician when EIPH is suspected in horses, especially when there are abnormal findings on percussion of the caudodorsal areas of the chest. PMID- 11901593 TI - Treatment of canine atopic dermatitis with zafirlukast, a leukotriene-receptor antagonist: a single-blinded, placebo-controlled study. AB - Zafirlukast and placebo were administered orally as individual agents to 20 dogs with atopic dermatitis. The pruritus was effectively reduced by at least 50% in 2/18 (11%) dogs that completed the trial with zafirlukast. Two dogs vomited after administration of the active drug. PMID- 11901594 TI - Minimum space allowance for transportation of swine by road. AB - Space allowance for animals in transit is a consistent concern in many countries that are developing codes of practice and regulations to assure humane treatment of food producing animals. The minimum space allowance requirements for a broad size range of swine in transit has not been well described or scientifically substantiated. A maximal loading pressure recommendation for pigs weighing from 5 to 250 kg was derived by a consultative process involving the swine transportation industry, animal welfare groups, and a literature review. The recommended maximal loading pressure under ideal conditions for swine loaded in groups can be described as a hoerl model y = (37.53)(0.9969)w(W0.5008), where y = loading pressure in kg body weight/m2 and W = average animal body weight in kilograms. PMID- 11901595 TI - An improved polymerase chain reaction assay for the detection of Tritrichomonas foetus in cattle. AB - An improved polymerase chain reaction test has been developed to detect Tritrichomonas foetus, the causative agent of trichomoniasis in cattle. The test amplifies a region of the 5.8S ribosomal RNA gene of T. foetus, and it is simple, sensitive, and specific when compared with traditional methods to examine field samples. PMID- 11901598 TI - Mortality associated with gastric ulceration in swine. AB - On a large swine operation, necropsy revealed that 39 of 146 (27%) pigs during one week in April and 37 of 137 (27%) pigs during one week in June died from hemorrhage due to gastric ulceration. Contributory factors may have been feed withdrawal one day per week and feeding finely ground pellets. PMID- 11901596 TI - Abomasal ulceration and abomaso-pleural fistula in an 11-month-old beefmaster bull. AB - An 11-month-old, beefmaster bull presented with anorexia and signs of respiratory disease. Physical examination, thoracic ultrasonography and radiography, and pleural fluid analysis indicated pericarditis and septic neutrophilic pleuropneumonia. Postmortem findings were abomasal adherence to the diaphragm, a fibrotic fistulous tract connecting the abomasum and pleural cavity, granulomatous abomasitis, granulomatous pericarditis, and fibrinonecrotic pleuritis. PMID- 11901597 TI - Behavior modification and pharmacotherapy for separation anxiety in a 2-year-old pointer cross. AB - Separation anxiety is a common behavioral problem in dogs. Treatment is based on developing a behavior modification protocol that gradually desensitizes and counter-conditions the dog to being left alone, by rewarding calm, relaxed behavior. Judicious use of pharmacotherapy can be a useful adjunct to a behavior modification program. PMID- 11901600 TI - Engineering revival plan generates optimism. PMID- 11901599 TI - Outbreak of Cysticercus bovis (Taenia saginata) in feedlot cattle in Alberta. PMID- 11901601 TI - Benchmarking must embrace best value. PMID- 11901603 TI - Trusts benefit from pre-engineered construction methods. PMID- 11901602 TI - A dream achieved. PMID- 11901604 TI - Modular theatres operate optimally. PMID- 11901605 TI - Providing quality speedily. PMID- 11901606 TI - New surgical wing for Monaghan General. Modular facility precisely meets requirements. PMID- 11901608 TI - Taking political responsibility for nursing's future. AB - House and Senate bills, H.R. 4387 and S. 1864, to develop and fund new initiatives for solving the nursing and nurse educator shortage, were passed shortly before the 107th Congress recessed for Christmas. These bills are now slated for compromise through a Congressional conference committee. The window of opportunity to influence the outcome of this process is short. To gain favorable political action that will improve our ability to care for patients and educate future nurses during this process, ACT NOW. PMID- 11901609 TI - Mitochondrial disorders: a clinician's primer. AB - Mitochondrial disorders represent an extremely challenging set of conditions, partly due to the complexities of the mitochondrial genetic system. Nurses must be prepared with knowledge about genetic conditions so that they can participate fully in early identification of affected individuals and assist them in accessing the most current genetic diagnostic and treatment therapeutics. This knowledge will be critical as nurses collect family history data, provide current and accurate genetic information and support to individuals and families, and refer to and collaborate with genetic specialists in their communities. PMID- 11901610 TI - Is gastric occult blood analysis affecting patient care delivery? AB - Two adult medical-surgical units in a large university hospital challenged the value of performing gastric occult blood testing in the acute care setting. As a result of their investigational findings, gastric occult blood testing was eliminated. Nursing time and patient care dollars were spent more efficiently, without compromising patient outcomes. PMID- 11901611 TI - Necrotizing fasciitis: pathophysiology and treatment. AB - Necrotizing fasciitis (NF) is a life-threatening bacterial infection causing necrosis of the fascia, underlying skin, and vasculature. NF spreads rapidly, making immediate diagnosis important for survival. Treatment may involve the administration of several broad-spectrum antibiotics, surgical debridement, and skin grafting. In the following two articles, the pathophysiology, medical management, and nursing care of patients are discussed. An in-depth model care plan illustrates the complexity of the disease and its treatment. PMID- 11901612 TI - Necrotizing fasciitis: a model nursing care plan. AB - Accurate assessment and timely interventions are critical in caring for patients diagnosed with necrotizing fasciitis. A case example helps nurses assess and intervene the numerous problems commonly experienced by patients with necrotic fasciitis. PMID- 11901613 TI - Electronic data, the Internet, and private practice. PMID- 11901615 TI - Preventing and treating meningococcal meningitis. AB - Meningitis is a bacterial or viral inflammation of the meninges. Three thousand cases are reported each year with a mortality rate of 5% to 40%. Socioeconomic factors play a role in meningitis and may affect the speed of diagnosis and subsequent initiation of treatment. Transmission and pathogenesis of the disease enables nurses to recognize signs and symptoms and implement treatment and prevention strategies. PMID- 11901616 TI - [Angioplasty for atherosclerotic lesions of the left main coronary artery]. AB - The authors analyze results of coronary angioplasty and stenting in patients with ischemic heart disease with a lesion of the left main coronary artery. Injuries of the left main coronary artery took place in 14 (0.74%) out of 1900 patients after coronary angioplasty. A detailed analysis of the operation technique, indications and contraindications for endovascular intervention is given. The results of balloon angioplasty with and without the following stenting are compared. PMID- 11901618 TI - [Extended and combined operations for lung cancer]. AB - On the basis of classificating components of the TNM system 1720 patients with "later" stages of the development of lung cancer were selected from 3000 patients with this disease operated upon in the clinic for many years. Specific features of performing extended and extended combined operative interventions, treatment at the postoperative period are described and substantiated. After their discharge from the hospital correspondingly 26% and 21.5% of the patients who had the IIIA stage of the disease are alive for more than 5 years, that convinces in expediency of their surgical treatment. PMID- 11901617 TI - [Immediate results of reconstructive surgery in end-stage dilated cardiomyopathy]. AB - First experiences with multistage surgical reconstruction of the left ventricle, heart valves and coronary arteries in patients with the end stage dilated cardiomyopathy are summarized. During the last three years operations have been made on 21 patients aged from 24 to 63, eight patients having idiopathic cardiomyopathy and 13 ischemic cardiomyopathy. They had markedly disturbed hemodynamics, ejection fraction of the left ventricle less than 30% and its diastolic diameter more than 70 mm. Most of the patients were candidates for heart transplantation. The Batista and Dor operations were made in combination with plastic operations on the mitral and tricuspid valves. In patients with ischemic heart disease myocardial revascularization was also performed. Four patients died after operation from arrhythmia and heart failure, the others' state had improved with less sizes of the heart and 10-12% greater ejection fraction. A conclusion was made that such operations were expedient. PMID- 11901619 TI - [The concept of abruption of pancreatic necrosis--key to the solution of the problem of destructive pancreatitis]. AB - The main criterion of severity of acute destructive pancreatitis (ADP) is the volume of pancreatonecrosis formed during the first 24 hours of the disease. A prognostic system is developed allowing to urgently detect the "risk group"- patients with the developing massive pancreatonecrosis by means of the assessment of clinical criteria. In the process of intensive therapy of the "risk group" patients there occurs arresting, "abruption" of the destructive process which greatly improves the results of treatment (the frequency of purulent complications decreases from 32% to 14%, lethality from 45% to 19%). Different methods of "abrupting" therapy are described, the best of them is a combination of curative plasmapheresis with the intraaortal perfusion of the splanchnic area with antienzymes, antihypoxants and antithrombotics. In the group of 17 patients with severe ADP treated by the method described suppuration was noted in 1 (6%) patient, and no lethal outcomes. The pressing problem is to increase the number of patients with severe ODP treated by the method of "abrupting" therapy as early as possible which can be achieved with the help of early hospitalization, with using prognostic systems and creation of a network of specialized pancreatological clinics with the corresponding equipment. PMID- 11901620 TI - [Role of spermatic vein phlebography and scleroembolization in treating varicocele and preventing its recurrence]. AB - The phlebograms and intravascular treatment of varicocele were performed in 406 patients aged from 10 to 60 years. Scleroembolization of the left internal spermatic vein was successfully used in 402 of them. Dilatation and retrograde flow through the right internal spermatic vein were recorded in 101 of 238 patients during transjugular catheterization. This investigation has confirmed high effectiveness of the method of the intravascular combined scleroembolization in treatment of varicocele. Using the transjugular access facilitates not only performing the left side intervention but represents the only efficient means to fulfill the right side phlebography of the spermatic vein and its obliteration. The strategy of using the bilateral intervention for the detection of retrograde contrast of the right internal spermatic vein considerably improves the results of treatment of varicocele and decreases risk of relapses. PMID- 11901622 TI - [Microbiological specifics in the course of suppurative inflammatory processes in soft tissues of patients with diabetes mellitus]. AB - The focal microflora of pyo-inflammatory processes has been studied in dynamics in 200 patients with diabetes mellitus. It was found that a longer and more severe course was due to the changed specific composition of causative agents and to the increased specific diversity of the latter in the process of pyo inflammatory progressing. It was established that the usage of the method of "close" wound management with the application of oxytocin inhibited the change of the causative agents in the course of the disease, led to a more rapid elimination of microorganisms from the pyo-inflammatory focus, to the less frequency of recurrent surgical interventions and shorter duration of courses of treatment in contrast to the results obtained with the other methods of treatment. PMID- 11901621 TI - [Mechanisms of the development of venous hypertension in patients with varicose disease]. AB - The examination of 539 patients with varicose disease was fulfilled with the help of ultrasonic (dopplerography, angioscanning), rentgenological (phlebography, lymphography), functional (rheovasography, photoplethysmography) methods. Two forms of venous hypertension were established: superficial and deep. Hemodynamic disorders in patients with the superficial form result from the formation of isolated blood refluxes in the superficial, perforant and profound veins. In patients with this form the muscular-venous pump has mild disturbance and is clinically manifested as a slowly progressing course of the disease. The basis for the development of the deep form is a severe valve incompetence of the profound veins with the formation of associated blood refluxes in all the elements of the muscular-venous pump. The latter completely looses its function which leads to a rapid lympho-venous decompensation. PMID- 11901623 TI - [Objective assessment of trauma severity in patients with combined injuries]. AB - The work presents an analysis of using scales VPH-P(MT) and VPH-P(SP) on the basis of a retrospective investigation of results of treatment of 268 patients with a combined injury of the abdomen. A conclusion is made that these scales have reserves for an improvement. The authors propose to supplement the scale VPH P(SP) with the index of the patient's age, and the scale VPH-P(MT)--with an index of the blood loss volume. PMID- 11901624 TI - [Diagnosis and treatment of iatrogenic and criminal injuries of the esophagus]. AB - An experience with treatment of iatrogenic and criminal traumas of the esophagus included 57 patients. There were 36 patients with iatrogenic traumas (perforation of the esophagus during diagnostic and curative manipulations and operations on the lungs and esophagus). Different kinds of operations were made depending on the localization of the injury. Three patients of this group died. Criminal traumas of the esophagus (wounds with cold steel) were treated in 21 patients, sutures on the esophagus were put in 19 of them, drainage--in 2 patients. All the patients recovered. Better results of treatment of the penetrating iatrogenic and criminal injuries of the esophagus can be achieved by timely diagnosis and operation using the proper volume and method. PMID- 11901625 TI - [Prophylaxis of recurrent multiple echinococcosis of the liver in children]. AB - The article presents an experience with treatment of 47 children aged from 3 to 14 with multiple echinococcosis of the liver. The authors have made a comparative analysis of the antiparasitic remedies and of a wide spectrum bactericidal solution of Betadin. The results of microscopic, histological and electron microscopic investigations have shown Betadin to be highly effective and harmless for the intraoperative prophylactics of recurrent echinococcosis. It is expedient to use Betadin for multiple echinococcal lesion of the liver in children. PMID- 11901626 TI - [Complex radiologic investigation for disorders of defecation in children]. AB - The work presents findings obtained by the radial examination of the anorectal area including routine methods such as irrigography and balloon proctography as well as modern technologies such as endorectal ultrasonography with a transducer of high resolution (7.5 MHz) and computed tomography in 250 children with more common syndromes in colonoproctology: constipation and incontinence of feces. Results of the endosonographic investigation of the normal sphincter apparatus in children are described. It is confirmed that endosonography is of great importance for the detection of the isolated intumescence of the anal sphincter. Computed tomography is also necessary for traumas of the anorectal area in children. PMID- 11901627 TI - [Anastomotic suture incompetence after transperitoneal resection of the rectum]. AB - Sphincter saving operations were made in 68.1% of 276 radical operations on the rectum, 85.6% of them being transperitoneal resections. The advantages of this operation are: less traumaticity, blood loss and postoperative lethality as well as saved function of the sphincter apparatus. Incompetence of the sigmorectal anastomosis sutures developed in 29 (18.6%) patients. The dependence of frequency of this complication on the sex and age of the patients, on the character of the disease and localization of tumor, on using suturing apparatuses, radiation treatment and unloading colostomy was studied. The distinctions found proved to be unreliable. Direct causes of the development of the complication are thought by the author to be technical defects of the surgery, disturbed blood supply and regenerative abilities of the sutured end of the gut. Recommendations are given how to prevent the development of the complication and of its more severe consequences. PMID- 11901628 TI - [Use of Pervomur for prophylaxis of infectious complications in elective surgeries of abdominal organs]. AB - After the experimental investigations performed, the 0.6-0.8% solution of Pervomur (as irrigation of the operation area or application) was used in 103 patients. As compared with the control group the intraoperative using of Pervomur allowed the frequency of postoperative complications to be reduced from 21% to 15% and the time of treatment was shortened. PMID- 11901629 TI - [Use of the new non-depolarizing myorelaxant, Nimbex, for anesthesiologic maintenance in high-risk thoracic operations]. AB - The article presents main results of the open randomized investigation of the effectiveness and safety of the new non-depolarizing myorelaxant Nimbex included in the composition of anesthesiological maintenance of thoracal operations of high risk. The authors give data of its influence on neuromuscular conduction and main hemodynamic indices during different periods of the operative intervention. The assessment of the intubation condition, depth of the neuromuscular block and of the dynamics of reestablishment of the neuromuscular conduction is shown. A conclusion is made of the possibility to use the myorelaxant in question as a preparation of choice for the maintenance of thoracal operations in high risk patients. PMID- 11901630 TI - [Assessment of adequacy of analgesia with spectral analysis of sinus rhythm variability in the maintenance of anesthesia]. AB - A spectral analysis of the sinus rhythm (5-minute groups of cardiointervals) was made in 32 patients during anesthesia by the method of a fast Fourier transform with the estimation of the spectral density in 3 conventional ranges (HF, LF, VLF). Its indices were shown to depend on the stages of anesthesiological maintenance and operative intervention. PMID- 11901632 TI - [Cholecystogenic cyst after mucoclasis of the gallbladder as a cause of postcholecystectomy syndrome]. PMID- 11901633 TI - [Transcatheter embolization in hemobilia after transcutaneous cholangitic drainage]. PMID- 11901634 TI - [An accessory pancreas in the wall of Meckel's diverticulum]. PMID- 11901631 TI - [New surgical methods for treating hepatic hydatic cyst]. AB - The authors have developed a set of instruments for endovideosurgical treatment of hydatid cysts of the liver. It was successfully used in operative ablation of the cysts by a traditional method in 9 patients, and in 7 patients laparoscopic echinococcus ectomy was performed without any complications. PMID- 11901635 TI - [Long-term complications of a foreign body in the small intestine]. PMID- 11901636 TI - [Acute strangulated colonic ileus (volvulus) in the early postoperative period after cesarean section]. PMID- 11901637 TI - [Role of enteral feeding in nutritional support of surgical patients]. AB - Main stages of artificial nutritional support (ANS) in the surgical clinic are considered as a scientific system of diagnostic and curative measures. Principle aspects of trophological screening of the surgical patient are elucidated. The algorithm of choice of the principle of ANS is presented: parenteral and enteral. Grounds are given to confirm the priority of the enteral method of ANS at the main stages of managing the surgical patient: before operation, during it, at the early and late postoperative periods. PMID- 11901638 TI - [Aleksei Nikolaevich Maksimenkov (1906-1968)]. PMID- 11901639 TI - [Current state of the problem of treating acute cholecystitis]. PMID- 11901640 TI - [Unfavorable effects of leukocytes in hemotransfusion in surgery]. PMID- 11901641 TI - What is the evidence for a causal link between hygiene and infections? AB - Even in an era in which access to personal "cleanliness" and a public health infrastructure are readily available in developed countries, illnesses associated with day care centres and homes continue to be a problem. The inhabitants of less developed countries, on the other hand, must contend with an inadequate public health infrastructure, lack of education programmes, and economic limitations in obtaining hygiene products. Therefore, less developed countries carry a greater burden of morbidity and mortality from infectious illnesses. The objective of this review is to examine and assess the epidemiological evidence for a causal relation between hygiene practices and infections. The Medline database was searched from January 1980 to June 2001 and studies were included if the outcome(s) was infection or symptoms of infection, and if the independent variable(s) was one or more hygiene measures. The strength of the association as measured by the relative reduction in risk of illness was appreciable and generally greater than 20%. Despite methodological strengths and limitations of the studies assessed, the weight of evidence collectively suggests that personal and environmental hygiene reduces the spread of infection. The results from this review demonstrate that there is a continued, measurable, positive effect of personal and community hygiene on infections. PMID- 11901642 TI - World War I may have allowed the emergence of "Spanish" influenza. AB - The 1918 influenza pandemic caused 40 million deaths, and so dwarfed in mortality and morbidity the preceding pandemic of 1889 and the 1957 and 1968 pandemics. In retrospect, much can be learnt about the source, the possible subterranean spread of virus, and the genetic basis of virulence. The World Health Organization has urged every nation to prepare a pandemic plan for the first global outbreak of the 21st century. We present an appraisal of epidemiological and mortality evidence of early outbreaks of respiratory disease in France and the UK in the years 1915 to 1917. Certain of these earlier focal outbreaks--called epidemic bronchitis rather than influenza--occurred during the winter months when influenza was known to be in circulation, and presented with a particular heliotrope cyanosis that was so prominent in the clinical diagnosis in the world pandemic outbreak of 1918-1919 (the Great Pandemic). The outbreaks in army camps at Etaples in France and Aldershot in the UK in 1916-1917 caused very high mortality in 25-35 year olds. Increased deaths from bronchopneumonia and influenza were also recorded in England. We deduce that early focal outbreaks of influenza-like disease occurred in Europe and on the balance of probability the Great Pandemic was not initiated in Spain in 1918 but in another European country in the winter of 1916 or 1917. We suggest that the pandemic had its origins on the Western Front, and that World War I was a contributor. PMID- 11901643 TI - Stage of the epidemic and viral phenotype should influence recommendations regarding mother-to-child transmission of HIV-1. AB - This article argues for a new approach to use of nevirapine in the prevention of vertical transmission of HIV-1. Existing antenatal surveillance should be strengthened to plan geographical allocation, and subsequent evaluation, of a "nevirapine plus" programme. As the epidemic evolves the programme should also and, ideally, care should be tailored to individual women. Underpinning this approach is evidence that a more virulent viral phenotype appears in many patients with advanced HIV-1 infection. This phenotype will become more common at the population level as the epidemic progresses. As efficacy of zidovudine correlates with viral phenotype, and use of the drug may alter phenotype, there is an urgent need for a replacement that is safe to use with nevirapine. PMID- 11901644 TI - Walter E Stamm--towards control of urinary tract infections. Interview by Pam Das. PMID- 11901645 TI - Anthrax update. PMID- 11901646 TI - Software forecasts tropical diseases. PMID- 11901647 TI - Pilot HIV care programme begins in Senegal. PMID- 11901648 TI - Vaginal vaccine for recurrent urinary-tract infections. PMID- 11901649 TI - Bacterial obsession. PMID- 11901650 TI - Jungle conceals Ebola origins. PMID- 11901652 TI - Can vaccines thwart the consequences of a bioterrorist attack? PMID- 11901653 TI - AIDS control in sub-Saharan Africa--are more drugs and money the solution? PMID- 11901654 TI - Resistance of Candida species to antifungal agents: molecular mechanisms and clinical consequences. AB - Candida albicans and related species pathogenic for man become resistant to antifungal agents, in particular triazole compounds, by expression of efflux pumps that reduce drug accumulation, alteration of the structure or concentration of antifungal target proteins, and alteration of membrane sterol composition. The clinical consequences of antifungal resistance can be seen in treatment failures in patients and in changes in the prevalences of Candida species causing disease. These effects were seen unequivocally in HIV-infected patients with oropharyngeal candida infections, but their incidence has decreased dramatically with the introduction of highly active antiretroviral therapy. The evidence for similar emergence of antifungal-resistant yeast strains and species in other types of candida infections is confounded by non-standardised susceptibility testing methods and definitions of a resistant fungal isolate. Recent large-scale surveys of yeasts isolated from blood cultures, based on standardised methodology and resistance definitions, do not support the view that antifungal resistance in pathogenic yeasts constitutes a significant or growing therapeutic problem. PMID- 11901655 TI - BCG--different strains, different vaccines? AB - After nearly a century of use, BCG vaccines continue to generate controversy and confusion. Their ability to prevent tuberculosis in studies has been inconsistent. When they have been protective, no clear mechanism of action has been established. Furthermore, the existence of different BCG strains has been described since the 1940s. These strains vary according to several laboratory properties, which may or may not translate into a discernible effect on vaccination. With recent genomic comparisons, it is now clear that different BCG vaccine strains have evolved and differ from each other and from the original BCG first used in 1921. Some of these genetic alterations explain certain variations in laboratory properties of BCG. However, these mutations in BCG strains have yet to be shown to affect BCG-associated protection and/or adverse effects. Continuing research is attempting to assess the effect of these genetic alterations on properties of BCG strains, with the goals of suggesting the ideal BCG for vaccination and providing avenues for improvement on existing BCG vaccines. PMID- 11901657 TI - Reflections on a rose: a story of loss and longing. AB - This case study discusses the psychotherapy of an older woman who presented with anxiety and depression decades after a heartbreaking loss in childhood. Maggie's story recalls a lifetime of longing and searching for her father, whose death was denied by those closest to her. Religious proscriptions contravened the nine-year old's accepting the reality of his death and understanding her own intense feelings. Masked, unresolved, complicated grieving punctuated her life. Maggie entered therapy as an adult confounded by death. "Creative integrationism" therapy included traditional grief work, guided imagery, journal writing, and poetry. As Maggie began to confront the deaths of loved ones, she explored the meaning of life and became more fully engaged in living. Several years after termination, Maggie revisited her journey in therapy and shared her reflections with the authors. This study illustrates the need for a grieving child to accomplish the tasks of mourning; the healing role of ritual; and the role of the psychiatric nurse to facilitate grieving even when the profound and unresolved loss comes to the fore after more than half a century has passed. PMID- 11901656 TI - Efficacy of highly active antiretroviral therapy in HIV-1 infected children. AB - Although the reduction in HIV-1-related deaths with highly active antiretroviral therapy (HAART) is similar in adults and children, the extent of the changes in two important surrogate markers HIV-1 RNA levels and CD4+ T cell counts, differs widely. In most paediatric studies virological response rates to HAART are inferior to those in adults. This review provides an overview of the paediatric clinical studies using HAART and seeks to improve the understanding of factors that may contribute to success or failure of HAART in children. An overview of all current articles on paediatric clinical trials using HAART is provided. 23 papers were available. HIV-1 RNA loads and CD4+ T cell counts were used as primary outcome measures. Virological response rates were highly variable, both among the different antiretroviral drugs but also among different studies using the same medication. Four studies in which dosages of the administrated protease inhibitor (PI) were adjusted after pharmacokinetic evaluation had superior virological response rates compared with those in which fixed dosages were used. Immunological response rates were more uniform than virological responses. In almost all studies increases of CD4+ T cell counts are reported independent of the extent of the virological response. Side-effects of HAART were generally mild, transient, and of gastrointestinal origin. Significant percentages of patients with serum lipid abnormalities were reported in three paediatric studies. However, signs of clinical lipodystrophy were not observed. The inferior virological response rates, which have been reported in HIV-1 infected children treated with HAART form a reflection of the challenges that are encountered in the treatment of these children. Difficulties with adherence and with the pharmacokinetics of PIs in children require an intensive, child-adjusted approach. A practical approach to therapy in institutions without tertiary care facilities may be induction therapy with a lopinavir containing regimen (lacking a need for therapeutic drug monitoring), to reduce high viral load levels followed by an easily tolerated maintenance regimen, for example containing abacavir or nevirapine. PMID- 11901658 TI - The enigma of severe mental illness: a Swedish perspective. AB - Today mental health professionals are challenged in supporting people with severe mental illness that live within their communities. The community treatment is, however, characterized by an uncertainty about how to best support them in their everyday lives, and professionals from different disciplines often have divergent opinions about the care. The aim of this study is to explicate the existential meaning of living with severe mental illness. Interviews with persons who relocated from an institutional setting to a community placement were analyzed within an interpretive approach. The results of the study found that people with severe mental illness experience an existential loneliness due to difficulties in changing previous suppositions about human relationships. They do not develop connections through shared new experiences with other people in their lives. One central implication of the findings is that because people with severe mental illness seem unable to benefit from new experiences, mental health nurses should consider relational aspects when planning, implementing, and evaluating nursing care. PMID- 11901659 TI - Post-polio syndrome: psychological adjustment to disability. AB - Although the Pan American Health Organization declared in 1995 that polio had been eliminated in the Western Hemisphere, life-altering effects of the disease continue for many survivors. It is known as Post-Polio Syndrome (PPS). The sheer number of individuals experiencing the symptoms has attracted the attention of the medical community. These physical symptoms are severe enough to change the quality of life and require lifestyle changes for people with PPS to cope with the disease. The psychological implications for individuals who must face the reemergence of a disease they thought they had defeated 30 to 40 years ago are staggering. Thus, there is a crucial need for health care professionals, especially mental health nurses and psychotherapists, to address mental health issues that individuals with PPS experience. PMID- 11901660 TI - Ethics and praxis: alternative strategies to physical restraint and seclusion in a psychiatric setting. AB - This descriptive article highlights a 42-month project in which a comprehensive program revision was implemented in a psychiatric hospital that included numerous alternative strategies to the use of patient restraint and seclusion. The results of this project include a 94% reduction in the rate of restraint and seclusion, development of extensive staff and patient education programs, and comprehensive programmatic alterations consistent with a paradigm shift emphasizing collaboration, empowerment, and ethical clinical practice. PMID- 11901662 TI - A plea for end-of-life content in the journal. PMID- 11901661 TI - Collaboration--together we can find the way in dual diagnosis. AB - Service systems in health and community agencies are struggling to deliver mental health services to adults with an intellectual disability. Many professionals feel ill equipped to assess and treat mental health disorders in this population. This Australian case study describes the collaborative effort required to meet the complex health needs of a client with an intellectual disability and the needs of her family, and the role played by a specialist, Disability Health Service. The key elements of this successful interagency collaboration are outlined and include good communication, adequate resourcing, and a willingness to resolve dynamic tensions and learn from each other. PMID- 11901663 TI - Keeping it together: how women use the biomedical explanatory model to manage the stigma of depression. AB - Although considerable research has been conducted on women who are depressed, the actual experiences and voices of women have not been central to this research. Therefore little is known about how women make sense of depression as they live with and manage it in their daily lives. Our purposes in doing this study were to (1) examine how women experience and manage depression and treatment, and (2) investigate the core components of women's explanatory models of depression (including beliefs about etiology, onset of symptoms, pathophysiology, course of illness, and treatment needs). We interviewed 43 women living in a small city in Western Canada who had sought treatment within the previous five years. Data were analyzed using the constant comparison method of grounded theory. In this paper we will focus on the core concept, Keeping it Together, and its three supporting categories, (1) Taking Up a Biomedical Explanation for Depression, (2) Using the Biomedical Explanatory Model (BEM) to Manage the Stigma of Depression, and (3) The Inadvertent Effects of Adopting a BEM. PMID- 11901664 TI - Increased prevalence of malignant diseases in the close neighborhood of children with cancer. AB - Clustering of cancer in families may be due to chance, inherited genetic mutations, common exposure to environmental agents, or a combination of these factors. The authors, to address a clinical impression that cancer occurs more often in the environment of a child with cancer, investigated whether the prevalence of cancer among children and adults in the neighborhood of children with cancer was higher than the prevalence in the neighborhood of healthy children. One hundred thirty-seven children diagnosed with a malignant disease between 1981 and 1992 at the Department of Pediatrics, University Hospital of Linkoping, Sweden, were investigated and compared with 232 healthy control children. The control children were traced from the official Swedish population registry. It was found that 13 percent of the children with cancer and six percent of the control children were close neighbors of other children diagnosed with cancer (p < .05). Cancer also was more common in the circles of acquaintances around the children with cancer than in circles of acquaintances around control children (p < .03). The frequency of cancer in the neighborhood or in the circle of acquaintances was significantly greater in older children than in younger children. These results support the hypothesis that environmental factors can initiate or precipitate cancer in children as well as in adults. PMID- 11901666 TI - Meeting employee expectations: exploring change through employee feedback. PMID- 11901665 TI - The effects of different environmental education programs on the environmental behavior of seventh-grade students and related factors. AB - This study used random allocation to separate out groups of students from four Taipei junior high schools, each of which underwent a different environmental education program, in order to examine the effects of such programs on students' environmental behavior and related factors. Results indicate that Taiwanese junior high schools should coordinate the teaching of environmental programs with other school activities to obtain the most ideal results. PMID- 11901667 TI - Risk factors for acute myeloid leukemia and multiple myeloma: a combination of GIS and case-control studies. AB - Risk factors for acute myeloid leukemia (AML) and multiple myeloma (MM) include exposure to toxic chemicals present in tobacco smoke, as well as to emissions from industrial operations and petroleum refinery waste dumps. The study reported here identified these risk factors among case patients and control patients in Orange County, California, from 1984 to 1993 and determined the significance of the risk factors in the study population. A case-control study was performed for 604 cases of AML and 643 cases of MM; there were 7,112 control subjects who had colon cancer. The model included the variables smoking history, occupational history, and residence in a census tract with a petroleum refinery waste dump. A geographic information system (GIS) analysis also was performed to correlate the incidence of AML and MM with proximity to the six dump sites that received large amounts of petroleum refinery waste. Current smokers were found to be at an increased risk of AML with an odds ratio of 2.0. Laborer/equipment cleaners and transportation workers/movers were at risk of AML with odds ratios of 3.5 and 2.4, respectively. Construction/resource extraction workers were at risk of MM with an odds ratio of 2.8. GIS analysis determined that the risk for MM was 1.6 cases per mile for 10 or more years of residence near a large chemical dump. The authors were able to identify census tracts with a high incidence of AML and MM, and to perform distance analysis using a statistical measure of spatial randomness. The case-control study identified occupational and lifestyle risk factors for AML and MM that were not apparent from census-tract-level data. PMID- 11901668 TI - Nonlinear kinetics: at the crossroads of chemistry, physics and life sciences. PMID- 11901669 TI - Investigation of nonlinear dynamical properties by the observed complex behaviour as a basis for construction of dynamical models of atmospheric photochemical systems. AB - The importance of the investigation of nonlinear dynamical properties (NDPs) of the atmospheric photochemical systems (PCSs) was demonstrated in ref. 1 and 2 (A. M. Feigin and I. B. Konovalov, J. Geophys. Res., 1996, 101 (D20), 26038; 1. B. Konovalov, A. M. Feigin and A. Y. Mukhina, J. Geophys. Res., 1999, 104 (D3), 3669). The only known way to study NDPs of any natural dynamical system (including atmospheric PCSs) is to construct a mathematical model of the system. The key point here is adequacy of the NDPs of the constructed model to the system observed. We propose a new approach to construction of such an adequate model for systems manifesting nonstationary chaotic behaviour and describe an algorithm based exclusively on nonlinear dynamical analysis of the observed time series (TS) without invoking any a priori knowledge about the properties of the system observed. Potentialities of the algorithm are demonstrated with the aid of a computer model of the mesospheric PCS. The duration of the "observed" TS is limited so that the system demonstrates only one--chaotic--type of behaviour, without any bifurcations throughout the observed TS. The proposed algorithm enabled us to make a correct prognosis of bifurcation sequences and calculate probabilities to reveal, at the time instant of interest, predicted regimes of the system's behaviour for times much greater than the length of the initial TS. PMID- 11901670 TI - A new chemical system for studying pattern formation: bromate-hypophosphite acetone-dual catalyst. AB - A modified version of the short-lived BrO3(-)-H2PO2(-)-Mn(II)-N2 oscillator, the BrO3(-)-H2PO2(-)-acetone-dual catalyst system, where the catalyst pair can be Mn(II)-Ru(bpy)3SO4, or Mn(II)-ferroin, or Mn(II)-diphenylamine, shows long lasting batch oscillations in the potential of a Pt electrode and in colour, accompanying periodic transitions between the oxidised and reduced forms of the catalysts. Experimental conditions for the oscillations are established. The origin of the batch oscillations and the role of the catalyst pair in the oscillatory behaviour are discussed. The new system is ideally suited to the study of waves and patterns in reaction-diffusion systems, since in addition to the longevity of its spatial behaviour in batch, it produces no gaseous or solid products and exhibits significant photosensitivity. PMID- 11901671 TI - Low-dimensional manifolds in tropospheric chemical systems. AB - Ordinary differential equations derived from large nonlinear systems of chemical reactions are computationally expensive to solve because of the large number of coupled species and the large range of time-scales present. The range of time scales often spans several orders of magnitude leading to stiff systems of equations requiring implicit numerical techniques. The use of slow manifolds for the description of long time-scale chemical processes has two advantages in that it reduces the number of variables required and also the stiffness of the chemical system by assuming that the fast time-scales are in local equilibrium with respect to the slower ones. The method exploits the existence of a low dimensional manifold within a large-dimensional species phase space onto which the system quickly collapses. This paper investigates the existence of slow manifolds for nonlinear tropospheric chemical systems and presents a simple method for estimating the local dimension of the manifold using linear perturbation theory. The method is demonstrated for several tropospheric mechanisms over diurnal simulations including a subset of the Master Chemical Mechanism describing butane oxidation and the formation of ozone in the troposphere. It is shown that the intrinsic dimension of the slow manifold varies diurnally and depends on photolytic processes and the relative concentrations of major pollutants. PMID- 11901672 TI - A numerical study of spatial structure during oscillatory combustion in closed vessels in microgravity. AB - The existence and spatial development of gas-phase, thermokinetic oscillations under the influence of mass and thermal diffusion have been investigated by numerical methods in a 1-dimensional system. The conditions correspond to those that would be experienced under microgravity. The interest arises because there have been recent experimental investigations of oscillatory reactions, involving cool flames during butane oxidation, as part of the NASA, KC135 microgravity flight programme. The Sal'nikov, thermokinetic scheme, which is a two-variable model representing an intermediate chemical species and reactant temperature (taking the form P-->A-->B), forms the basis of the present work. In this model, thermal feedback occurs through the exothermicity of the second step and the non linearity is derived from its temperature dependence. There are no known chemical examples that satisfy Sal'nikov's formal structure but Griffiths and co-workers conceived an experimental analogue under terrestrial conditions whereby a gaseous reactant was allowed to flow from an external reservoir into a closed, heated reactor at a controlled rate via a capillary tube which fed the reactant to the centre of the vessel. The exothermic reaction that occurred in the vessel satisfied the necessary conditions for the second step and the inflow, with no temperature dependence, represented a physical analogue to the first step of the Sal'nikov scheme. Thermokinetic oscillations were observed and the range of conditions for their existences was investigated. One of the experimental systems was the exothermic reaction between hydrogen and chlorine. To represent the Sal'nikov conditions hydrogen was fed slowly into the reactor, which already contained chlorine. We have exploited this chemical system and its experimental implementation in the present paper to investigate the behaviour when no convection or bulk gas motion occurs and when heat and mass transport is driven solely by diffusion. We study the response of alternative numerical approaches to the way in which the first step of the scheme is simulated. In the first, the precursor (P) is supplied at the same rate simultaneously throughout the cells representing the reactor. This is close to the concept of the Sal'nikov model. In the second method, a fixed rate of supply is applied at the inner boundary of the axisymmetric, 1-dimensional system. This is analogous to the experimental procedure. The numerical results show how oscillatory states can be sustained as a result of heat and mass transport by diffusion. The temporal and spatial evolution of reaction in a range of circumstances is discussed. PMID- 11901673 TI - Spatial bifurcations of fixed points and limit cycles during the electrochemical oxidation of H2 on Pt ring-electrodes. AB - Pattern formation during the oscillatory oxidation of H2 on Pt ring-electrodes in the presence of electrosorbing ions was studied under potentiostatic control for three different positions of the reference electrode (RE). The position of the RE crucially affects the degree of the global feedback which is imposed by the potentiostatic operation mode, and the three configurations selected corresponded to zero, maximum and intermediate global coupling. In the absence of global coupling, 'communication' among different positions occurs exclusively through migration coupling (the electrochemical counterpart to diffusion in reaction diffusion systems). In this case, spatially inhomogeneous oscillations that were attributed to a spatial bifurcation of the homogeneous limit cycle were observed throughout. This implies that the system is Benjamin-Feir unstable. For the strongest global coupling adjustable, travelling pulses were found that emerged in a wave bifurcation with n = 1 from the homogeneous steady state. The pulses exhibited modulations in velocity and width that most likely resulted from the interaction between inhomogeneities of the catalytic surface and the nonlinear reaction dynamics. In the case of an intermediate global coupling strength, a diversity of spatio-temporal motions was observed. The dynamics ranged from pulses over target patterns and so-called asymmetric target patterns to mixed states where two or three of these states alternate. For some parameters these mixed states were in addition separated by bursts of the system to a nearly homogeneous unreactive state. PMID- 11901674 TI - Identification of the intermittency-I route to chaos in oscillating CO oxidation on zeolite-supported Pd. AB - For the oscillating oxidation of CO on a zeolite-supported palladium catalyst the transition to chaos could be observed in a very narrow region of the CO concentration in the feed. The reaction was carried out under the conditions of a continuous stirred tank reactor. A careful choice of the method for time series analysis led to the unambiguous identification of the intermittency-I route to chaos in the catalytic system despite the rather limited number of data points which can be acquired under normal pressure conditions. The route to chaos could be derived from the variation of the Fourier spectrum and the Poincare section as a function of the CO concentration in the feed. The embedding dimensions for the observed chaotic attractors of dE > or = 10 are much higher than the embedding dimensions obtained during UHV single crystal studies. High embedding dimensions indicate that the dynamic behaviour of the system has to be simulated with a distributed model which describes the collective behaviour of many Pd particles in the zeolite crystallite. PMID- 11901675 TI - HPLC analysis of complete BZ systems. Evolution of the chemical composition in cerium and ferroin catalysed batch oscillators: experiments and model calculations. AB - In the last few years many new reaction routes and intermediates have been discovered in the mechanism of the Belousov-Zhabotinsky (BZ) reaction with the aid of high performance liquid chromatography (HPLC). These previous HPLC studies, however, were limited to the Ce(4+)-organic substrate (malonic or bromomalonic acid) systems only. Very recently some measurements were made on a cerium catalysed full BZ system but only in its induction period. The present work follows the evolution of the main chemical components in a cerium and in a ferroin catalysed full BZ system from the start until the end of the oscillatory regime in a batch reactor. While recording the potential oscillations of a bromide selective electrode we measured from time to time the concentration of the following components: malonic and bromomalonic acids and bromate as main components; malonyl malonate, ethanetetracarboxylic and bromoethenetricarboxylic acids which are recombination products of organic free radicals; oxidized intermediates: tartronic, oxalic (OA) and mesoxalic (MOA) acids, and brominated products: dibromoacetic and tribromoacetic acids. Recombination products are generated in the intervals when the autocatalytic reaction is "switched off". In the course of the autocatalytic periods, however, the organic radicals react with the inorganic bromine dioxide radical mainly which leads to the formation of MOA and OA. Due to a very fast Ce(4+)-MOA reaction, MOA can be detected in the ferroin catalysed BZ system only. Our model calculations deal exclusively with the cerium catalysed system. The suggested new Marburg-Budapest-Missoula (MBM) model includes both negative feedback loops (bromous acid-bromide ion Oregonator type and bromine dioxide-organic free radicals Radicalator type feedback) and the recently discovered radical-radical recombination reactions. Comparison of the experimental data with the model calculations shows a good qualitative agreement but some open problems still remain. To overcome these problems oxygen atom transfer and other redox reactions are proposed. PMID- 11901676 TI - Oscillatory dynamics protect enzymes and possibly cells against toxic substances. AB - We have used the oscillating peroxidase-oxidase (PO) reaction as a model system to study how oscillatory dynamics may affect the influence of toxic reaction intermediates on enzyme stability. In the peroxidase-oxidase reaction reactive intermediates, such as hydrogen peroxide, superoxide, and hydroxyl radical are formed. Such intermediates inactivate many cellular macromolecules such as proteins and nucleic acids. These reaction intermediates also react with peroxidase itself to form an inactive enzyme. The fact that the PO reaction shows bistability between an oscillatory and a steady state gives us a unique possibility to compare such inactivation when the system is in one of these two states. We show that inactivation of peroxidase is slower when the system is in an oscillatory state, and using numerical simulations we provide evidence that oscillatory dynamics lower the average concentration of the reactive intermediates. PMID- 11901677 TI - pH oscillations in the hemin-hydrogen peroxide-sulfite reaction. AB - Oscillatory behaviour in the pH value has been observed during the oxidation of sulfite by hydrogen peroxide mediated by hemin, a well known enzyme model compound, in a continuous-flow stirred tank reactor. The dynamics of this reaction has been studied for a variety of flow rates of the reactants. As the flow rates increase, the oscillations evolve from relaxation oscillations to more complex shapes, displaying, among others, bursting behaviour. A reaction mechanism is proposed that involves the autocatalytic oxidation of HSO3- by H2O2, while slow equilibria between different pH-dependent forms of hemin account for the feedback loop which gives rise to oscillatory dynamics. It is shown in experiments that no participation of CO2 is required for oscillations to occur. PMID- 11901678 TI - Control of the excitability of neuronal tissue by weak external forces. AB - The spreading depression (SD) is a pronounced example of excitation-depression waves in excitable media, to which neuronal tissue according to its structure and functions belongs. SD waves can especially easily be observed in the vertebrate retina which is neuronal tissue and a true part of the central nervous system (CNS). According to the high intrinsic optical signal (IOS) concomitant with the retinal spreading depression (rSD), it can be monitored with standard video imaging techniques, thus the retina has been used in our studies as a suitable model system for neuronal tissue in general. In particular, the control of wave set-up and propagation in excitable media by weak external forces is of high interest. Accordingly, the interaction of rSD waves with DC and AC electromagnetic fields of low amplitude and frequency and with gravity has been investigated in this study. The dependence of rSD-wave propagation velocity on the given parameters as one important indication of excitability control has been investigated in detail. Our results with rSD waves are partially compared to another well known excitable medium, the Belousov-Zhabotinsky reaction, where some data about the effects of electrical fields and gravity have already been published. PMID- 11901679 TI - Spatio-temporal dynamics in glycolysis. AB - During the glycolytic degradation of sugar in a thin layer of yeast extract, travelling waves of NADH and protons can be generated that carry a state of high enzymatic activity through the system. The controlled initiation of such waves with an activator of the enzyme phosphofructokinase (PFK) and the influence of various salts and co-factors on the propagation dynamics are investigated. Furthermore a first study of the dispersion of waves is presented. The experimental characterisation of this in vitro system contributes to unravelling the possible role of glycolysis for biological information processing. In this context, the provision of chemically available energy in the absence of compartmentation by glycolysis is of primary importance. PMID- 11901680 TI - Synchronization of glycolytic oscillations in a yeast cell population. AB - The mechanism of active phase synchronization in a suspension of oscillatory yeast cells has remained a puzzle for almost half a century. The difficulty of the problem stems from the fact that the synchronization phenomenon involves the entire metabolic network of glycolysis and fermentation, and consequently it cannot be addressed at the level of a single enzyme or a single chemical species. In this paper it is shown how this system in a CSTR (continuous flow stirred tank reactor) can be modelled quantitatively as a population of Stuart-Landau oscillators interacting by exchange of metabolites through the extracellular medium, thus reducing the complexity of the problem without sacrificing the biochemical realism. The parameters of the model can be derived by a systematic expansion from any full-scale model of the yeast cell kinetics with a supercritical Hopf bifurcation. Some parameter values can also be obtained directly from analysis of perturbation experiments. In the mean-field limit, equations for the study of populations having a distribution of frequencies are used to simulate the effect of the inherent variations between cells. PMID- 11901681 TI - Complex morphogenesis of surfaces: theory and experiment on coupling of reaction diffusion patterning to growth. AB - Reaction-diffusion theory for pattern formation is considered in relation to processes of biological development in which there is continuous growth and shape change as each new pattern forms. This is particularly common in the plant kingdom, for both unicellular and multicellular organisms. In addition to the feedbacks in the chemical dynamics, there is then another loop linking size and shape changes with the reaction-diffusion patterning of growth controllers in the growing region. In studies by computation, the codes must incorporate, alongside the usual solvers of the partial differential dynamic equations, a versatile growth code, to express any kind of shape change. We have found that regulation of shape change in particular ways (e.g. to make narrow-angle branchings) demands new features in our chemical mechanisms. Our growth algorithm is for a surface growing tangentially, but moving outward and changing shape to accommodate the extra area. This is potentially applicable both to the tunica layer of multicellular plant meristems and to the growing tip of the cell surface, e.g. in the morphogenesis of single-celled chlorophyte algae which display branching processes: whorl formation in Acetabularia (Dasycladales) and repeated dichotomous branching in Micrasterias (Desmidiaceae). For computational studies, a hemispherical shell is a reasonable idealization of the initial shape. We describe results of two types of study: (1) Pattern formation by three reaction diffusion models, with contrasted nonlinearities, on the hemispherical shell, particularly to find conditions for robust formation of annular pattern or pattern for dichotomous branching, both of which are common in plants. (2) Sequential dichotomous branchings in a system growing and changing in shape from the hemispherical start. PMID- 11901682 TI - Chemical waves in open flows of active media: their relevance to axial segmentation in biology. AB - The boundary forcing of open flows of active media can lead to a variety of spatiotemporal structures, depending on the local kinetics of the medium and on the characteristics of the forcing. Here, we demonstrate that regardless of the local kinetics, the combination of flow and boundary forcing is a powerful method for replacing intrinsic modes with extrinsic ones. This entrainment of dynamics has important implications for biological morphogenesis. During early embryonic development it is frequently observed that stripes of gene expression and segments arise one after the other along a growth-axis. We show that axial growth can be viewed as an open flow of cells away from a growth zone. Based on this realisation, we demonstrate using three generic reaction-diffusion-advection schemes how a space-periodic structure is induced, one "segment" at a time along the growth/flow axis, by a segmental clock that is synchronised within the growth zone. The schemes are investigated in the context of an abrupt and a gradual change in the properties of the segmental clock. Experimental observations provide evidence that the latter is involved in the early development of many vertebrates. PMID- 11901683 TI - Excitability in chemical and biochemical pH-autocatalytic systems. AB - Using two different kinds of pH systems--the papain catalyzed hydrolysis of N benzoyl-L-arginine ethyl ester in a membrane reactor and the bromate-sulfite ferrocyanide (BSF) reaction in the CSTR--we study the relation among excitability, oscillations and bistability, and the ability of the system to respond to external periodic perturbations. Excitable properties of dynamical systems are examined in terms of a threshold set which is used to characterise dynamics in the reactor subject to external periodic stimuli. A precise definition and a method of calculating the threshold set are formulated. Two kinds of excitability distinguished by either direct or indirect initiation of the activatory process are found in both pH systems. Periodic pulsed perturbations of the BSF system display a nontrivial dependence of an excitation number on the forcing period. We examined this system also in oscillatory mode by looking at the phase shifts caused by single-pulse perturbations and constructing the phase transition curves (PTCs). PMID- 11901684 TI - Spatial bistability and waves in a reaction with acid autocatalysis. AB - The phenomenon of spatial bistability has recently been proposed for a comprehensive understanding of a number of chemical patterns observed in open spatial reactors consisting of thin films of gel diffusively fed from one side. We study experimentally and numerically this phenomenon in the tetrathionate chlorite reaction characterized by an acid superautocatalysis. We focus on the similarities and differences with previous studies on the chlorine dioxide-iodide reaction. In addition, we show that this reaction, which is only bistable in a continuous stirred tank reactor, can exhibit oscillatory and traveling waves when diffusion comes into play. Our computations suggest that the nonstationary behaviour originates from differential diffusive transport. PMID- 11901685 TI - Pattern formation and spatial self-entrainment in bistable chemical systems. AB - We describe the formation of spatial structures generated by diffusive instabilities in bistable systems. The coupling between the different spatial modes emanating from the two homogeneous steady states can then give rise to self parametric instabilities favoring the occurrence of resonant rhombic or quasiperiodic structures such as superlattices or quasicrystalline patterns. PMID- 11901686 TI - Turbulent fronts in resonantly forced oscillatory systems. AB - Phase fronts in the forced complex Ginzburg-Landau equation, a model of a resonantly forced oscillatory reaction-diffusion system, are studied in the 3:1 resonance regime. The focus is on the turbulent (Benjamin-Feir-unstable) regime of the corresponding unforced system; in the forced system, phase fronts between spatially uniform phase-locked states exhibit complex dynamics. In one dimension, for strong forcing, phase fronts move with constant velocity. As the forcing intensity is lowered there is a bifurcation to oscillatory motion, followed by a bifurcation to a regime in which fronts multiply via the nucleation of domains of the third homogeneous phase in the front. In two dimensional systems, rough fronts with turbulent, complex internal structure may arise. For a critical value of the forcing intensity there is a nonequilibrium phase transition in which the turbulent interface grows to occupy the entire system. The phenomena we explore can be probed by experiments on periodically forced light sensitive reaction diffusion systems. PMID- 11901687 TI - Experimental and theoretical studies of feedback stabilization of propagating wave segments. AB - Experimental and theoretical studies of the excitability boundary for spiral wave behavior are presented. The boundary is defined by unstable wave segments, which are stabilized by using a negative-feedback control algorithm. A kinematic description of the constant-size, constant-shape wave segments is presented. PMID- 11901688 TI - Effects of non-ionic micelles on transient chaos in an unstirred Belousov Zhabotinsky reaction. AB - The behaviour of the Ce(IV)-catalyzed Belousov-Zhabotinsky (BZ) system has been monitored at 20.0 degrees C in unstirred batch conditions in the absence and presence of different amounts of the non-ionic micelle-forming surfactants hexaethylene glycol monodecyl ether (C10E6) and hexaethylene glycol monotetradecyl ether (C14E6). The influence of the non-ionic surfactants on both the kinetics of the oxidation of malonic acid (MA) by Ce(IV) species and the behaviour of the BZ reaction in stirred batch conditions has also been studied over a wide surfactant concentration range. The experimental results have shown that, in unstirred batch conditions, at surfactant concentrations below the critical micelle concentration (c.m.c.) no significant change in the dynamics of the Belousov-Zhabotinsky system occurs. Beyond this critical concentration the presence of micelles forces the BZ system to undergo a chaos-->quasi-periodicity- >period-1 transition. Thus, the surfactant concentration has been considered as a bifurcation parameter for a Ruelle-Takens-Newhouse (RTN) scenario. Addition of increasing amounts of non-ionic surfactants has no significant effect on the kinetics of the reaction between MA and Ce(IV), but it influences the oscillatory parameters of the stirred BZ system. At surfactant concentrations below the c.m.c. all the oscillatory parameters are practically unaffected by the presence of surfactant, while beyond this critical value the induction period is the same as in aqueous solution but both the oscillation period and the duration of the rising portion of the oscillatory cycle decrease. In all cases, the experimental trends have been ascribed to the enhancement in the medium viscosity due to the presence of micelles. PMID- 11901689 TI - Control and coupling of spiral waves in excitable media. AB - We report the complex dynamics of spiral waves observed in the ferroin-catalyzed BZ reaction. The reaction is run in an open unstirred reactor (CFUR) with the catalyst immobilized on a polysulfone membrane. The catalyst-loaded membrane is placed between two well stirred compartments which are fed with solutions of sulfuric acid/malonic acid/bromide and sulfuric acid/bromate, respectively. An electrical field perpendicular to the membrane can be applied via Pt-ring electrodes or, alternatively, via transparent electrodes made of ITO-coated glass. In the field-free case relatively simple target and spiral patterns are observed in the membrane. If an alternating electrical field is applied the spiral core drifts through the membrane. The actual trajectory of the spiral tip depends on the amplitude and frequency of the applied electrical field. If the perturbation parameters are chosen properly the wave fronts break up and new spiral cores emerge under the influence of the alternating field. Complex spatio temporal patterns may be induced which are reminiscent of "spiral-chaos". After switching off the perturbation the system returns to its previous, "simple" behaviour. Our experimental observations are confirmed by model calculations based on the Barkley model of spiral waves. The technique of using modulated excitability to control the dynamics of spiral waves is further extended to the coupling of two spirals in two CFURs. We present numerical simulations based on two identical excitable reaction-diffusion (RD) systems which are mutually coupled. The coupling is based on the observation of an arbitrarily chosen point inside each of the RD systems: If a chemical wave passes the point of optical observation in system 1 an electric field is applied to system 2 and vice versa. Thus, a local observation made in one system is transformed in a global perturbation of the second CFUR. We report the observation of CFUR states where the two spiral waves are spatially and temporally coupled to each other. PMID- 11901690 TI - Nonlinear behaviour of simple ionic systems in hydrogel in an electric field. AB - The stationary behavior of ionic reaction-transport systems contained in hydrogel located between two reservoirs of electrolytes is investigated. The effects of the applied voltage, the composition of the electrolytes in the reservoirs and the distribution of fixed charge in the hydrogel on the spatial patterns of charge, electric potential, temperature and concentrations of the individual components are studied systematically by mathematical modelling. Such systems, containing only several ions (for example, Cl-, K+, OH- and H+) can function as electrolyte diodes and transistors and can exhibit oscillations and hysteresis. The mathematical description of the studied systems is based on balances of species, enthalpy and charge, on Poisson's equation and on the finite rate description of the water dissociation/recombination reaction, without assuming local electroneutrality. The modeling results are compared with experiments, including the system with hydrogel connecting reservoirs of strong base and acid studied by Noszticzius and co-workers. [L.H. Hegedus, N. Kirschner, M. Wittmann and Z. Noszticzius, J. Phys. Chem. A, 1998, 102, 6491 (ref. 1); L. H. Hegedus, N. Kirschner, M. Wittmann, P. Simon and Z. Noszticzius, Chaos, 1999, 9, 283 (ref. 2).] PMID- 11901691 TI - Self-oscillating polymer chain in a laser field. AB - We have investigated the dynamical behaviour of the rhythmic conformational change between the folded compact state and the unfolded state in a single polymer chain under thermodynamically open conditions. It is shown that the spontaneous rhythmic change in the conformation of a single polymer chain (T4DNA, 166 kbp, contour length: 56 microns) is generated using a focused continuous wave (CW) Nd:YAG laser beam (wavelength lambda = 1064 nm), where the focused laser beam plays a dual role, both trapping a polymer chain at the focus and creating a temperature gradient around the focus. Furthermore, the whole process of the rhythmic conformational change: the course of melting, nucleation and growth between the folded and unfolded states has been clarified. The rhythmic change in the conformation is discussed in terms of the limit-cycle oscillation driven by the dissipation of the photon energy. PMID- 11901692 TI - [Endovascular dilatation in the treatment of patients with obliterating atherosclerosis of main arteries of lower extremities]. PMID- 11901693 TI - [Infrequent localization of rupture of abdominal aortic aneurysm]. PMID- 11901694 TI - [Post-traumatic hepatitis]. PMID- 11901695 TI - [Indications for hypoxitherapy]. AB - Hypoxytherapy is the non-medicinal therapeutic method using gaseous hypoxic mixture (GHM) with decreased oxygen contents. The method is based on the activation of body protective mechanisms, phagocytosis stimulation, microcirculation improvement, sedative effect. GHM therapy is indicated in neurosis, CHD, hypertension, chronic pulmonary obstructive disease, to prevent from side effect of ionizing radiation, to increase the resistance during complex therapy of oncologic patients. The method is contraindicated in acute diseases, decompensation of chronic diseases. The authors noted that it is reasonable to use GHM low doses in rehabilitation period after acute pneumonia and in geriatrics. Quite satisfactory effect was obtained in therapy of lower extremity atherosclerosis. For the first time the fact of concrement passage under GHM influence was registered and hypoxytherapy was included into the complex therapy of urolithiasis. PMID- 11901696 TI - [Structural alterations in erythrocyte membranes in patients with myocardial infarction]. AB - The patients with myocardial infarction have marked changes in the structure of both protein and lipid parts of the membrane. These changes are connected with destruction of antioxide cell protection against the background of developing hypoxia and "oxide stress". In such patients the increase in HbA2 and HbF factors may be regarded as the adaptive mechanism of hypoxia conditions that develop due to decrease in myocardial propulsive ability and deterioration of blood rheological properties. Change of structural and functional characteristics of erythrocyte membranes leads to increase in their hardness that is connected with the increase in blood viscosity, pressure upon the impaired myocardium and extension of necrotic zones. Similar structures of erythrocyte and cardiomyocyte membranes allow to draw a conclusion about the presence of similar disorders in the latter membranes. It shows the necessity to introduce into the treatment scheme of myocardial infarction patients the drugs inhibiting lipid oxidation. PMID- 11901697 TI - [Endoscopic videosurgery in a gynecologic department of a military hospital]. PMID- 11901698 TI - [Medical support of the personnel of check and observation posts in the zone of Georgian-Abkhazian conflict]. AB - During the operations for peace support by collective forces it is necessary to deploy check (CP) and observation (OP) posts usually at a distance of dozens kilometers from each other and from the battalion base camp. Depending on CP and OP dislocation place, problems they should solve the number of servicemen in the posts can vary from 12 to 50. For rendering medical care the company base post has the permanent medical instructor of the company and in the other CPs and Ops- supernumerary medical instructors of the posts from the number of the trained servicemen. The experience of medical support of collective forces for peace support in the zone of Georgian-and-Abkhazian conflict allowed to summarize and outline some ways of its improvement which first of all are directed to more effective work of medical instructors. PMID- 11901699 TI - [Characteristics of sympathetic adrenal responses in marginal arterial hypertension]. PMID- 11901700 TI - [Evaluation of effectiveness of therapeutic paraffin-containing girdle "Physomed" in patients with urolithiasis]. PMID- 11901701 TI - [Use of vaccine "Euvacs B" in a large polyclinic hospital]. PMID- 11901702 TI - [Eye condition in operators of visual display terminals]. PMID- 11901703 TI - [Development of military medical expertise in Moscow Military District]. PMID- 11901704 TI - [Higher medical school within the walls of Moscow military hospital]. PMID- 11901705 TI - [Establishment of psychiatric services for servicemen during combat operations]. PMID- 11901706 TI - Carcinoid tumor. AB - Carcinoid tumors are slow growing malignancies which occur most frequently in the gastrointestinal tract (about 74%). They can also be found in the bronchus, ovary, lung, thymus, kidney or thyroid gland. Carcinoid tumors are usually identified histologically by their affinity to silver salts, or more specifically by immunocytochemistry using antibodies against their specific cellular products. Survival rates depend on the location of primary tumor, extent of locoregional and metastatic disease, functional status of the tumor and the feasibility of complete surgical extirpation. Clinical manifestations are often vague or absent. Nevertheless, tumours secrete bioactive mediators which may in approximately of 10% of patients engender various elements of characteristics of carcinoid syndrome. Patients with advanced carcinoid disease should be treated with aggressive medical and surgical therapies. (Ref. 103.) PMID- 11901707 TI - Mechanisms of replication of alpha- and betaherpesviruses and their pathogenesis. AB - The diseases caused by herpes simplex virus (HSV) and human cytomegalovirus (CMV) differ and distinct differences in biological properties of these viruses can be noticed at laboratory work. Despite of this, the structure of DNA and the replication cycle of both viruses shows remarkably common features. Analogous proteins encoded by both viruses, act at initiation of viral DNA transcription, at viral DNA synthesis, at nucleocapsid formation and envelopment. On other hand, considerable differences occur during maturation of virions and at their egress from infected cells. Both viruses in question developed strategies to escape immune recognition by cytotoxic T cells and/or to interfere with the antibody response. Both viruses are widespread in human population and are able to establish latency. Finally, their prevention and/or prophylaxis by effective vaccines has not been solved. Recently, the significance of both viruses has increased. HSV2 is an important pathogen acquired by sexual contact, while CMV reactivates under immunosuppression (post-transplantation, tumours, combined activation in the presence of human immune deficiency virus) and/or causes congenital infection. Chemotherapy of HSV mediated diseases seems more effective than that of CMV mediated infection, because the CMV inhibitor ganciclovir is much more toxic than the CMV inhibitor acyclovir and its derivatives. (Tab. 6, Fig. 5, Ref. 52.) PMID- 11901709 TI - Thirty two years old patient with adult Still's disease. AB - Adult Still's disease is characterized by diverse clinical and laboratory findings, which may lead to errors in the differential diagnosis, and possible injury of the patient's health due to wrong therapeutic management. In the following case report, we describe a case of a 32-year old patient with fever of unknown etiology. The final diagnosis of Morbus Still adultorum was determined five months after his first check-up. The course of the disease was complicated by acute hepatitis caused by drug toxic damage. We have applied immunosuppressive therapy with very good clinical and laboratory responses. (Ref. 9.) PMID- 11901708 TI - Adaptive changes of antioxidant status in development of experimental diabetes. AB - BACKGROUND: In clinical and experimental research, attention is paid to the role of antioxidant defense systems in the prevention of diabetic complications. Little information is available about regulation of the endogenous level of antioxidants in the state of chronic oxidative stress in relation to the development of diabetes, and particularly about coenzyme Q as one of the most important endogenous antioxidants with an irreplaceable function in mitochondrial bioenergetics. PURPOSE: To examine changes in concentrations of two important lipophilic antioxidants, coenzyme Q9 and alpha-tocopherol, in rat tissues during the development of experimental diabetes. METHODS: Experimental diabetes in male Wistar rats, was induced by streptozotocin in the dose of 55 mg/kg intravenously. Coenzyme Q9 (CoQ9) and alpha-tocopherol (alpha-toc.) were determined in myocardial and skeletal muscles and in kidney tissue after 1, 6 and 8 months of diabetes duration by the method of high-performance liquid chromatography. RESULTS: Myocardial CoQ9 content increased progressively in the course of diabetes development by 14, 29 and 61%, while in skeletal muscles and kidney, the increases were not dependent on the duration of diabetes. The content of alpha toc. increased in the myocardium after 8 months of diabetes duration, in kidney tissue and skeletal muscles, it did not change in comparison with control rats. CONCLUSIONS: An increased content of the lipophilic antioxidants coenzyme Q9 and alpha-tocopherol in tissues of diabetic rats is regarded as an adaptation of the antioxidant defense system to chronic oxidative stress. The exact mechanisms of accumulation of these antioxidants in diabetic tissues could be elucidated by studies investigating their relation to changes in lipid content and to the total of bioenergetic and antioxidant capacities. (Fig. 3, Ref. 25.) PMID- 11901710 TI - Modulation of evoked potentials by interfering mental activity. AB - BACKGROUND: Non-specific influences (attention, mental set, mental imagery) take part in modulation of information processing in the human central nervous system at various levels of central hierarchy. OBJECTIVES: To broaden knowledge concerning the conjunction of motor control and cognition as well as to discuss some possible psychophysiological mechanisms. METHODS: Somatosensory evoked potentials, motor related cortical potentials and the saccadic eye movement related potentials were recorded over frontal and parietal cortices in the control condition and compared with those registered under the influence of a mental imagery task introduced as a modulatory factor. Special focus was devoted to their early components. MAIN RESULTS: Early components of the above potentials, which are assumed to represent the coding of physical characteristics of evoking stimuli, are changed under the influence of mental activity. Frontal evoked potentials are markedly modulated as compared to those recorded parietally. In general, the results support the view that the preparation for both, real movements and mentally simulated movements share probably some common mechanisms at the cortical level. CONCLUSIONS: Modulatory effects of mental activity upon the used evoked potentials point to the possible participation of the central gating mechanism, attention focussing and some other psychophysiological mechanisms related to internal representation of sequence of events. The results suggest that experimental conditions need to be exactly specified and examined subjects must be properly instructed. (Fig. 3, Ref. 12.) PMID- 11901711 TI - Health--mental health--quality of life. AB - BACKGROUND: Three positive aspects of human life manifestations are most frequently referred to as different, but closely interrelated concepts: health, mental health and quality of life (QoL). OBJECTIVES: Discussion of most frequently presented (WHO) definitions of the above mentioned concepts and of their certain shortcomings. METHODS: Critical review of concepts and definitions. MAIN RESULTS: The definition of health stresses the functional aspects of all life manifestations of man, i.e. biological, psychological, and social. The functional qualities of mental health are characterised by five areas. QoL is discussed as a broad concept pertaining to the set of material, biological, psychological, social and cultural needs and demands related to the well-being and life satisfaction of an individual. CONCLUSIONS: All of the discussed concepts point to the great complexity of factors playing their roles in human health, mental health, and the QoL. They all require an integrating and/or integrated concept as to the optimal character of biological, psychological, social, and cultural manifestations of human life. (Ref. 23.) PMID- 11901712 TI - Sensitive markers of the repolarization alterations in systemic hypertension. AB - INTRODUCTION: The early phase of essential hypertension has been associated with changes in cardiovascular regulation caused by imbalance in some parts of autonomic nervous system. Autonomic effect of various stimuli on haemodynamic variables is usually tested by changes in blood pressure (BP) and/or heart rate (HR). It is known that increased sympathetic drive of ventricles can interfere with repolarization process. THE AIM: This study was focused on reactive changes of maximal spatial T vector (sTmax) and R-R intervals, in relation to BP changes in 79 boys and men, averaged age 17 +/- 2 yrs. 36 from them where adolescents with elevated BP (high normal or hypertension I according to WHO/ISH1999) (EBP), 19 normotensives (NBP), and 24 normotensive sportsmen. R-R intervals and maximal spatial T vector were recorded by a PC (Cardiag METE, Prague) with Frank lead system while sitting in mid-respiratory position, during mental arithmetic (MA) and during passive head-up tilting to 60 degrees. BP was measured simultaneously by a cuff sphygmomanometer, using phases 1 and 5 of Korotkoff sounds. RESULTS: MA resulted in significant BP increase in all subgroups, however the reactive changes of systolic BP as well as magnitude of R-R shortening, and sTmax decline were about two times higher in sportsmen. Head-up tilting evoked in all subgroups a significant increase of diastolic BP that was again higher in sportsmen vs. EBP and NBP. R-R interval became significantly (p < 0.02) and more often shortened in sportsmen than in EBP and NBP. Relatively more evident decline of sTmax (more than 20%) in EBP was the highest change from all observed parameters to both tested stimuli. CONCLUSION: Our results indicate that for analysis of the effect of stimuli, which modulate balance in autonomic nervous system, it is helpful to pay attention also to the parameters of repolarization process that may represent a sensitive indicator of sympathetic tonization in myocardial ventricles. (Tab. 4, Fig. 2, Ref. 27.) PMID- 11901713 TI - Transtrochanteric rotational osteotomy, section after Sugioka. AB - BACKGROUND: Femoral caput necrosis is an actual therapeutic problem, because it appears mainly in mid-aged people. The necrotic focus is most often localized in the proximal and ventral parts of the femoral bone capitulum, which, from the biomechanical point of view is the most loaded part. These cases can be possibly treated by transtrochanteric rotational osteotomy. By means of this operation we rotate the necrotic focus to the less loaded part of the joint in correlation with the acetabulum. PURPOSE: The aim of this paper is to inform about a rarely performed operation after Sugioka: transtrochanteric rotational osteotomy in coincidence with avascular necrosis of the femoral bone caput. We would like to point to the possibility of a joint-saving operation, which enables to postpone implantation of total endoprosthesis to older age. METHODS: We analyzed the operational therapy performed at our department in the period 1998-2000, focusing especially on joint-saving operations due to femoral caput necrosis. We turned our attention to transtrochanteric rotational osteotomy after Sugioka. In four casuistics, we describe the results of these operations. RESULTS: Transtrochanteric rotational osteotomy was performed in 12 patients. The treatment is described in four casuistics. The results after 1-3 years are good. The illness has not progressed in any of patients, pain has regressed, mobility improved and it was not necessary to implant total endoprosthesis. CONCLUSION: We consider the transtrochanteric rotational osteootomy, section after Sugioka, as one of the possible treatments in mid-aged patients. Our effort is to perform joint-saving operations in order to postpone the implantation of total endoprosthesis. (Fig. 10, Ref. 13.) PMID- 11901714 TI - Media. A look behind the scenes. PMID- 11901715 TI - Retention and recruitment. Leaving by the back door. PMID- 11901716 TI - Why MS nurses need to multiply. PMID- 11901717 TI - Technology. Brave new world? PMID- 11901718 TI - Would the grass be any greener? PMID- 11901720 TI - Roses are red, bruises are blue. PMID- 11901719 TI - Are nursing students really so badly off? PMID- 11901721 TI - Can hoist use degrade a patient? PMID- 11901722 TI - Address your stress. PMID- 11901723 TI - 'How could someone like me have a nervous breakdown?'. PMID- 11901724 TI - NHS in crises. What's goin' on? PMID- 11901726 TI - Is the NSF causing gaps in care? PMID- 11901725 TI - Stroke: a group learning approach. PMID- 11901728 TI - A support initiative for nursing homes. PMID- 11901727 TI - Log on to develop practice. PMID- 11901729 TI - HIV and AIDS. 3. Treatment and care issues. PMID- 11901730 TI - Violence at work. A&E staff say trust neglects safety. PMID- 11901732 TI - Long-term care. Health Which? Survey reveals poor staff checks. PMID- 11901731 TI - Infection control. Naming MRSA trusts could alarm patients. PMID- 11901733 TI - Whistle-blowing. Second hearing for nurse's unfair dismissal claim. PMID- 11901734 TI - Child protection. Baseline training is not enough. PMID- 11901735 TI - Public health. Stick to advising MMR, nurses told. PMID- 11901736 TI - Violence at work. Staff fear for their lives as patients riot. PMID- 11901737 TI - Let's work together to create an integrated leg ulcer service. PMID- 11901738 TI - Evaluating the cost and efficacy of leg ulcer care provided in two large UK health authorities. AB - OBJECTIVE: This prospective study investigated the cost and efficacy of leg ulcer care over a three-month period during 1993, 1994 and 1999. It compared two health authorities (Stockport and Trafford) whose populations totalled 540,000. METHOD: All patients with active leg ulcers were invited to community leg ulcer clinics offering research-based innovations in care. Patients without significant arterial disease (ankle brachial pressure index less than 0.8) were treated with multilayer compression bandaging. RESULTS: The 42% healing rate reported in the original Stockport study was maintained at 40% in 1999, although the 65% healing rate achieved by the leg ulcer clinics in 1993 was not replicated, with rates falling to 46%. Following the opening of community leg ulcer clinics in Trafford, healing rates rose from 20% to 42%. The annual expenditure on leg ulcer care in Stockport increased from 65,545.56 Pounds to 83,344.30 Pounds, while in Trafford the cost of care dropped from 151,375.35 Pounds to 53,176.76 Pounds between 1994 and 1999. CONCLUSION: This study suggests that reductions in costs and improvements in healing rates can be sustained in a mature community leg ulcer clinic programme staffed by specialist leg ulcer nurses. PMID- 11901739 TI - Why do some cavity wounds treated with honey or sugar paste heal without scarring? AB - As well as having antimicrobial properties, honey and sugar paste are associated with scarless healing in some cavity wounds. This article uses evidence to suggest why these products can modify excessive collagen production to prevent scarring. PMID- 11901740 TI - ABPI Dopplers and DVT. PMID- 11901741 TI - Fistula management following an appendicectomy: nursing challenges. AB - Preservation of skin integrity, nutritional support and removal of odour are key features of fistula management. This case study describes the care of a patient with a fistula wound that discharged copious amounts of semi-solid faecal matter. PMID- 11901742 TI - Identifying protein production in wound healing: current techniques. AB - Proteins regulate would healing. Knowledge of the proteins present in poorly healing wounds may reveal new targets for treatment. This article describes how proteomics can be used to help identify the proteins involved in would healing. PMID- 11901743 TI - Cost-effectiveness of treating deep diabetic foot ulcers with Promogran in four European countries. AB - OBJECTIVE: This study aimed to estimate the cost-effectiveness of treating non superficial diabetic foot ulcers with Promogran plus good wound care (GWC) compared with GWC alone in four European countries (France, Germany, Switzerland and UK). METHODS: An existing Markov-based health economic model of non superficial diabetic foot ulcers was adapted to incorporate the relative efficacy of Promogran compared with GWC alone as demonstrated in a randomised controlled trial. Treatment with Promogran was modelled for a maximum of three months. Country-specific treatment costs were used to estimate the incremental cost per ulcer-free day gained over 12 months. Some parameter assumptions were changed to assess the sensitivity of the results. RESULTS: Within the first three months of treatment, 26% of ulcers in the Promogran cohort healed compared with 20.7% in the GWC cohort. Over the 12 months, the average number of months spent in the healed state was 3.41 (GWC) and 3.75 (Promogran). Promogran treatment was found to be cost-saving in all four countries, using year 2000 Euro values. CONCLUSION: Promogran with GWC may be cost-effective, perhaps even cost-saving, under a wide variety of assumptions for the treatment of neuropathic foot ulcers. DECLARATION OF INTEREST: This study was funded by Ethicon Gmbh (Johnson and Johnson), Germany. PMID- 11901744 TI - Nurses' views about pain and trauma at dressing changes: a central European perspective. AB - OBJECTIVE: This study investigated nursing and medical practitioners' perceptions of pain and trauma at dressing change in three German-speaking European countries: Austria, Germany and Switzerland. It follows a similar study by Hollinworth and Collier conducted in the UK. METHOD: A total of 3300 questionnaires were posted to practitioners in the three countries. All of the practitioners had attended at least one educational event on wound management organised by the researchers. The questionnaire contained closed questions about the participants' experience of dressing change and pain, their place of work and their freedom to select wound management products. RESULTS: A 15.1% response rate was achieved. The main aim at dressing change was to prevent trauma to the wound (30%) and to prevent infection (29%) and pain (21%). Dressing removal (51%) and wound cleansing (41%) were cited as the phases of dressing pain most likely to cause pain. Factors perceived to cause pain during dressing changes included dressings that adhere to the wound area (35%), particularly direct adherence to the wound (29%) and dried-out dressings (28%). Techniques used to prevent pain included rehydrating dressings. The responses also pointed to insufficient knowledge about low-adherent or non-adherent dressings among some respondents. CONCLUSION: The respondents perceived dressing removal and wound cleansing to be the most painful wound-care interventions. Their key objectives were to prevent pain and trauma at dressing changes. Lack of information on low- and non-adherent dressings needs to be addressed. DECLARATION OF INTEREST: This study was funded by Molnlycke Health Care AG, Switzerland. PMID- 11901745 TI - Clinical governance. 2: Obtaining consent in wound care: what are the key issues? AB - When undertaking a therapeutic intervention, healthcare professionals must ensure that they receive informed consent from the patient in order to protect both the individual and themselves. PMID- 11901746 TI - Can individualised nutritional support improve healing in therapy-resistant leg ulcers? AB - OBJECTIVE: This study investigated whether an individually designed programme of nutritional support can improve healing in otherwise therapy-resistant venous leg ulcers. METHOD: Six primary health-care patients, aged between 79 and 93 years, with venous ulcers that had been open for one year or more (range: 1.5-8 years) were recruited into the study. The patients were asked to follow an individualised diet plan which included the use of liquid dietary supplements. Ulcer area, anthropometric and biochemical variables, and energy and nutrient intake were assessed before intervention and then regularly for nine months. RESULTS: At nine months ulcer healing had occurred in two patients, of whom one had had ulcers on both legs. In a third patient the ulceration on one leg had healed and that on the other leg had almost healed. In a fourth patient, the ulcer area reduced by approximately 90%. CONCLUSION: The use of nutritional support might have assisted the wound healing in these patients. Although the relationship between nutritional supplementation and wound healing is not well defined, an appropriate nutritional plan is recommended if undernourishment is suspected and leg ulcers are not healing. PMID- 11901747 TI - Tissue viability programmes at the University of Hertfordshire. AB - Good practice requires an in-depth understanding of wound management and tissue repair, as well as critical-thinking skills. These education programmes give students an opportunity to develop these qualities. PMID- 11901748 TI - Management of a heavily exuding, painful wound with necrotising subcutaneous infection. AB - Wounds with necrotising fasciitis are often malodorous and produce copious exudate. Selecting appropriate dressings can alleviate these symptoms and improve the patient's quality of life within a short time period. PMID- 11901749 TI - Wound-bed preparation. PMID- 11901750 TI - Burn wound management in patients with epilepsy: adopting a multidisciplinary approach. AB - Many burns that occur following an epileptic seizure are deep due to prolonged contact with the thermal source. Primary care staff need to be aware of this and ready to refer patients to a burns unit. PMID- 11901751 TI - An exploration of patients' understanding of leg ulceration. AB - OBJECTIVE: This study aimed to identify patients' understanding of leg ulceration by examining their knowledge of the disease process and their expectations of their treatment outcomes. It also aimed to identify their attitudes to any patient information provided to them. METHOD: A total of 101 patients (median age: 75 years, range: 23-91; 54% female) with current leg ulceration of 16 months' median duration (range: 1-480) were asked about their understanding of their condition and treatment and their expectations of patient information provided by health-care professionals. RESULTS: The majority of patients believed they knew the cause of their ulceration (66%), most frequently attributing it to trauma (28%). The most important part of the treatment was felt to be bandaging (27%) and visiting the clinic (12%). Thirty-nine percent did not know what the term 'venous' meant. 'Trauma' was frequently described as a psychological problem, with only 7% associating the word with a knock or wound. Over half of the patients (64%) expressed an interest in acquiring further information, particularly on how they could assist ulcer healing (51%). CONCLUSION: For patient education to be effective, it must be tailored to the patients' vocabulary using simple concepts and straightforward, unambiguous messages. PMID- 11901752 TI - DVT: the forgotten factor in leg ulcer prevention. PMID- 11901753 TI - Clinical governance. 1: Evidence-based practice. AB - Practitioners have a responsibility to ensure their practice is based on sound clinical evidence and that the care delivered is of a high quality. What are the best ways of achieving this in the reality of the modern NHS? PMID- 11901754 TI - The cellular and molecular mechanism of CD4/CD8 lineage commitment. AB - A unique feature of alpha beta T-cell development is the central role played by clonally distributed T-cell receptors (TCR), which are encoded by somatically rearranged gene segments that produce a diverse, non-germline encoded set of receptors. Fate determination in individual T-cells is mediated by ligand receptor signals that arise from unprogrammed genetic interactions, under conditions in which the relevant ligand concentration and the receptor affinity are not evolutionarily controlled. A precursor T-cell with a TCR that either fails to demonstrate appreciable self-reactivity or binds with high affinity to reasonably abundant self-peptide major histocompatibility complex (MHC)-ligands will undergo apoptosis. In contrast, a precursor T-cell that shows lower affinity to moderately abundant ligands will receive suitable signals for survival and maturation. Recently, we have developed a rapid in vitro two-step organ culture system that permits homogeneous populations of non-transformed precursor T-cells to undergo selective commitment to the CD4 or CD8 lineage. Using this model, we have shown that the choice of positively selected ab T-cells between the CD4 helper and CD8 cytotoxic lineages is regulated by the TCR signaling duration in response to self-peptides bound to the MHC. PMID- 11901755 TI - Effects of constitution, atraumatic vertebral fracture and aging on bone mineral density and soft tissue composition in women. AB - Constitution, atraumatic vertebral fracture and aging affect bone mineral density (BMD) and soft tissue composition. The high body weight of obese women involves a high mechanical load being exerted on weight bearing bones compared with thin women, which probably contributes to their higher BMD and the lower incidence of fractures in obese women compared with thin women. Atraumatic vertebral fracture (AVF) is a typical osteoporotic fracture and its favorite site of AVF is the vertebral bodies of the thoracolumbar region. The BMD of weight bearing bones is lower in patients with AVF than in patients without AVF, whereas there is no significant difference in soft tissue composition between the two. The regional and total BMD decrease with advancing age. The magnitude of the decrease in lumbar and thoracic BMD is high compared with other regional BMD, and total fat mass and total lean mass decline with age to their respective minimal level. The high rate of decrease in lumbar and thoracic BMD appears to be due to the high content of trabecular bone compared with other regional bones. PMID- 11901756 TI - Antimutagenicity of Murdannia loriformis in the Salmonella mutation assay and its inhibitory effects on azoxymethane-induced DNA methylation and aberrant crypt focus formation in male F344 rats. AB - An 80% ethanol extract of Murdannia loriformis, a Thai medicinal plant, was examined for antimutagenic activity and cancer chemopreventive activity. In the Salmonella mutation assay, the extract showed antimutagenicity against 2-amino-3 methylimidazo [4,5-f]quinoline, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline, 2 amino-3,8-dimethylimidazo [4,5-f]quinoxaline, 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine, 2-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole, 3 amino-1-methyl-5H-pyrido[4,3-b]indole, 2-amino-6-methyldipyrido [1,2-a:3',2'-d] imidazole, 2-aminodipyrido[1,2-a:3',2'-d]imidazole, 2-aminoanthracene, 2-(2 furyl)-3-(5-nitro-2-furyl) acrylamide, N-methyl-N'-nitro-N-nitrosoguanidine and methylazoxymethanol acetate and reduced their mutagenicities to 31.4-67.9% at the dose of 10 mg/plate. However, it did not inhibit the mutagenicities of 2-amino-9H pyrido[2,3-b]indole, 2-amino-3-methyl-9 H-pyrido[2,3-b]indole, benzo[a]pyrene,N ethyl-N'-nitro-N-nitrosoguanidine and 1-nitropyrene. The extract itself showed no mutagenicity. The chemopreventive activity of M. loriformis was examined using azoxymethane (AOM)-induced aberrant crypt focus (ACF) formation in the colon of F344 rats. The extract at doses of 0.1-1.0 g/kg wt significantly inhibited ACF formation in the initiation stage (21-51%), although it was more effective at a lower dose. In the post-initiation stage, the extract also tended to inhibit ACF formation (12-27%) and significantly decreased the number of larger ACFs that have more than 3 aberrant crypts per focus. The extract inhibited the formation of O6-methylguanine and N7-methylguanine in the colonic mucosa and muscular layers but not or increased in the liver. These results indicate that M. loriformis extract has antimutagenic activity toward various known mutagens and that it inhibits AOM-induced ACF formation both in the initiation and post initiation stages in the rat colon. PMID- 11901757 TI - Distribution of calbindin-D28K immunoreactive neurons in rat primary motor cortex. AB - Distribution of calbindin-D28K immunoreactive cells in the primary motor area of the adult rat neocortex was studied in the present experiment. In the primary motor cortex, calbindin-D28K immunoreactivity was found in two populations of cortical neurons. One was composed of neurons heavily labeled with anti-calbindin antibody, which were present in two bands corresponding to cortical layers II III, and V. The morphological types of these cells were varied; they had oval, fusiform or mutiangular somata. The proximal dendrites of the heavily stained cells showed that these cells were non-pyramidal neurons, and they were either bitufted or multipolar cells. The other was a weakly stained population, mainly concentrated in layers II and III, that also contained pyramidal neurons. In addition, one outstanding feature of the neuropil staining deep to layer II was the labeling of the long, vertically oriented bundles of immunoreactive processes. Such a distinct pattern of calbindin-D28K immunoreactive neurons in the primary motor cortex suggests a relatively high density of calcium channels exists in the superficial layers of the rat primary motor cortex. PMID- 11901758 TI - Natural history of extruded lumbar intervertebral disc herniation. AB - We studied the natural history of extruded lumbar intervertebral discs using MRI. Forty-nine patients with lumbar disc herniation were included in this study. Ages ranged from 19 to 57. On the T2-weighted sagittal MR image, the signal intensity in the herniated mass was measured and the ratio to that in the original nucleus (i.e., nucleus pulposus from which they extruded) was calculated (signal intensity ratio; SIR). The relationship with SIR and duration of illness was evaluated. In ten patients who were re-examined by MRI after conservative treatment, the size of the herniation measured by T1-weighted axial MR image was compared before and after treatment. The signal intensity of HNP became higher than that of the original nucleus immediately following herniation and thereafter decreased with time, suggesting that initial hydration of the HNP occurred shortly after herniation followed by dehydration of the HNP. The size of the HNP with a SIR value of 1.2 and higher on T2-weighted MR images decrease with time, however, the HNP with a SIR below 1.2 did not show any size reduction. The SIR of 1.2 and higher is a good indicator predicting spontaneous reduction of the HNP. Dehydration in the HNP may play an important role in the reduction of the lumbar disc herniation. PMID- 11901760 TI - Traits of irrational beliefs related to eating problems in Japanese college women. AB - This study focused on the relation of irrational beliefs and Body Mass Index (BMI) to inappropriate eating attitudes in Japanese college women. A total of 110 nonclinical subjects completed the Japanese Irrational Belief Test (JIBT) and the Japanese version of the Eating Attitudes Test (EAT). The JIBT subscale of 'self expectation' had significant positive correlations with the EAT total score and the subscales of 'obsession with eating', 'dieting' and 'obese-phobia'. The JIBT subscale of 'dependence' had a significant positive correlation with the EAT subscale of 'obsession with eating'. BMI score showed significant positive correlations with the EAT total score and the subscales of 'dieting' and 'obesephobia'. The present results suggest that characteristic irrational beliefs are associated with inappropriate eating attitudes, suggesting that clarifying and then modifying the irrationality may be a useful method of preventive intervention in nonclinical young women with eating problems. PMID- 11901759 TI - Two Y-chromosome-specific polymorphisms 12f2 and DFFRY in the Japanese population and their relations to other Y-polymorphisms. AB - This study of male-specific genetic markers in the Japanese population was carried out in an attempt to refine the existing theories concerning its population genetics and migration events. We examined the relation between the constructed haplotypes of three biallelic Y-chromosome-specific markers (YAP, 47z and SRY) and the results of studying two other Y-specific polymorphisms of both 12f2 and DFFRY markers. The 12f2 marker was completely absent in 14.7% of Japanese males; all of them were haplotype II males. None of the Japanese males from other haplotypes or other East Asian populations showed any deletion of 12f2. In all haplotype II Japanese men, we found that DFFRY gene harbors a (C- >T) substitution polymorphism that was not found in any other population of this study. These results suggested that although haplotype II Japanese males share with the other haplotype II men from different geographical areas in having the YAP insertion on their Y-chromosomes, their Y-chromosomal structure is somewhat characteristically different. They are probably descendants of the ancestral Jomonese population who lived in Japan before the Yayoi immigrants entered Japan approximately 2300 years ago. These findings suggested that linkage studies between Y-specific markers are helpful in understanding the migratory patterns in East Asia. We also suggested that Japanese males have characteristically different Y-chromosomes compared with other populations. PMID- 11901761 TI - Linkage between prostate cancer incidence and different alleles of the human Y linked tetranucleotide polymorphism DYS19. AB - We studied the allele frequency distribution of the Y-chromosome linked tetranucleotide polymorphic microsatellite locus DYS19 in 90 prostate cancer Japanese patients from both Tokushima University hospital (Tokushima) and Saint Marianna University hospital (Kawasaki), Japan, comparing them to 99 matched male controls. Y-chromosomes from Japan as well as others from different geographical regions worldwide showed the five different alleles (A-E) with sizes varying from 186-202 bp, respectively. Comparison between DYS19 allelic frequency distribution among Japanese patients with prostate cancer and that of normal controls revealed significant differences regarding susceptibility or resistance to prostate cancer. We found that males with allele C of DYS19 are more susceptible to develop prostate cancer than males with other alleles (p = 0.02). The Odds Ratio was 2.04 with a 95% confidence interval (0.75-2.42), compared with males having other alleles. In contrast, males with the D allele of DYS19 were less exposed to prostate cancer than other males (p = 0.002); the Odds Ratio was 0.26 with a 95% confidence interval of (0.65-3.71). These findings support our hypothesis that male descendants from different Y-chromosomal origins are different regarding their susceptibility or resistance to develop prostate cancer (as a male-specific cancer). PMID- 11901762 TI - Angiostatic effects of corticosteroid on wound healing of the rabbit ear. AB - Wound healing is a complex biologic process with initial inflammation, granulation tissue formation, and matrix remodeling. We observed the relation between angiostatic effects and corticosteroid administration time in the rabbit ear chamber. Angiogenesis in the chamber was studied using a microscope television system. Two experiments were undertaken to represent the systemic and the topical administration of steroids. In experiment 1, 10 mg of triamcinolone acetonide was injected three times intramuscularly (on the day of implantation of the chamber, and the 7th and 14th day after implantation). Vascularization in this group was significantly delayed at the 7th, 14th, and 21st days but no difference from controls was observed in the size and density of vessels after its completion. In experiment 2, 3 mg of triamcinolone acetonide was injected once into the skin adjacent to the chamber on the 10th day after installment of chambers or on the day of installment. In the former group, new vascular growth was delayed until the 21st day after installment. The hemorrhagic zone had narrowed and vascular dilation was observed. In the latter group, endothelial budding was delayed and vascular constriction occurred. New vascular growth was severely delayed and granulation filling of the chamber was not completed. These results suggest not only that the topical administration had the stronger inhibitory effect on neovascularization than the systemic administration but that the effect differed depending on the stage of wound healing. In view of this effect of this steroid, we should pay careful attention to the time when steroids are administered to patients. PMID- 11901763 TI - Effect of ursodeoxycholic acid on azoxymethane-induced aberrant crypt foci formation in rat colon: in vitro potential role of intracellular Ca2+. AB - The studies were conducted to examine the precise nature of the suppressive effect of ursodeoxycholic acid (UDCA) on colonic aberrant crypt foci (ACF) formation. Fischer 344 rats were treated with a single dose of azoxymethane (AOM) (20 mg/kg, s.c.) and fed basal diet (MF) supplemented with UDCA (0.4%) during an initiation or a post-initiation stage. ACF were enumerated at the 2nd, 5th and 8th weeks after AOM administration (15-18 rats/group). The number of ACF in the UDCA treated group was decreased significantly in the initiation and post initiation stages at the 2nd (P < 0.01, P < 0.0001) and 8th weeks (P < 0.001, P < 0.0001), respectively, compared with untreated controls. In the time-course experiments, the effect of continuous feeding of UDCA (0.4%) on ACF formation was evaluated. ACF number was decreased significantly (P < 0.005) until the 16th week. UDCA showed a significant dose-dependent suppression of ACF number from a range of 0.1-0.4% UDCA. To approach the subcellular mechanisms of the effect of bile acids, the intracellular free Ca2+ concentration ([Ca2+]i) of bile acid treated rat colonic cancer cells (ACL-15) was examined. DCA and CDCA, which are promotive on ACF formation, induced a rapid increase in [Ca2+]i, while UDCA and CA, which are suppressive or non-effective on ACF formation, did not. These findings suggest that the promotive effect of bile acids may involve intracellular Ca2+ signaling. PMID- 11901764 TI - Genetic modification of dendritic cells and its application for cancer immunotherapy. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs). DCs pulsed with peptides of tumor-associated antigens (TAA) and tumor lysate have been used in cancer immunotherapy. An early clinical study demonstrated the safety of the use of DCs, but the clinical response was not sufficient. The gene modification of DCs with TAA and soluble factor genes such as cytokine and chemokine genes has been examined to enhance the antigen-presenting capacity of DCs. Viral vectors including retroviruses and adenoviruses have been reported to be useful to obtain a sufficient transduction efficiency into DCs. TAA gene transduced DCs could have several advantages compared with TAA peptide-pulsed DCs as follows: 1) The use of TAA gene-modified DCs are not restricted by MHC haplotypes. 2) The gene transduction with TAA genes is likely to present the unknown TAA peptides on DCs. 3) The gene-modified DCs show the prolonged presentation of TAA peptides. The transduction of DCs with cytokine genes including IL-12 and GM-CSF have also been reported to argument the antitumor effects of DCs. Although the results in the experimental systems were promising, the clinical application of gene-modified DCs includes several problems such as the standardization of methods of manipulation and gene-transduction of DCs. Approaches to solve them require further studies. PMID- 11901766 TI - Personal identification from skeletal remain by D1S80, HLA DQA1, TH01 and polymarker analysis. AB - The completely reduced skeleton was found in a mountain stream. We presumed that the skeleton was a woman from a morphology feature of the skull and the pelvis bone. The level of the suture of the skull indicated that the age was the first half of the 70-years old from the 60-years old. As a result of the police investigation, the possibility of 66 years old woman who was missing for about six months was suspected. To inquire into her identification, a skull and left thighbone were cut off, and blood was collected from the suspect's daughters. We examined blood (ABO) and DNA types (D1S80, HLA DQA1 TH01 and polymarker system) for the skeleton and the suspect's families. Blood and DNA types analysis of two daughters revealed that their patients have 19 alleles in 9 blood and DNA types. Twelve alleles were admitted in the skeletal remain among presumed 19 alleles. There was no blood and DNA type to deny the mother and daughter relation all of nine types. The skeletal remain was not contradicted from the above-mentioned result though thought daughters' mother. PMID- 11901765 TI - Interleukin (IL)-12 gene transduction and its functional expression into human bronchial epithelial cells (BEAS-2B) by adenovirus vector. AB - Interleukin (IL)-12 is known as a cytokine that augments the Th1 type response. Especially in allergic diseases such as a bronchial asthma, IL-12 induced restoration of the balance of the Th1/Th2 type immune response is an attractive strategy. In this study, the functional properties of the human bronchial epithelial cell line (BEAS-2B) transduced by an adenoviral vector encoding the human IL-12 gene were examined. Adenovirus vectors, AxCAegfp and Ax1CIhp40ip35 were transduced into BEAS-2B cells. Wild and gene-transduced BEAS-2B cells were incubated and the concentrations of IL-12 and IFN-gamma produced by co-cultured lymphocytes in the supernatant were measured using ELISA. The expressions of surface adhesion molecules, such as CD54 and CD106 were analyzed using flow cytometry. The efficiency of transgene expression of BEAS-2B cells was in a multiplicity of infection (MOI)-dependent manner and at an MOI of 30, the efficiency was approximately 80%. The gene-modified BEAS-2B cells produced biologically active IL-12 in dose- and time-dependent manners. IL-12 gene transduction did not significantly affect the expression of adhesion molecules (CD 54, CD106 and HLA-A,B,C) by BEAS-2B cells. These results suggest that the IL 12 gene may be successfully transduced into human bronchial epithelial cells by adenoviral vector to express IL-12 activity in vivo. PMID- 11901767 TI - Constructing the human dance of meaning. PMID- 11901768 TI - Ambiguity and hope: disclosure preferences of less acculturated elderly Mexican Americans concerning terminal cancer--a case story. PMID- 11901769 TI - Reading futility: reflections on a bioethical concept. PMID- 11901770 TI - What "race" cannot tell us about access to kidney transplantation. PMID- 11901771 TI - Denying culture in the transplant arena: technocratic medicine's myth of democratization. PMID- 11901772 TI - Do genetic relationships create moral obligations in organ transplantation? PMID- 11901773 TI - Adults are not big children: examining surrogate consent to research using adults with dementia. PMID- 11901774 TI - Equal access to cloning? PMID- 11901775 TI - Moral conundrums in the courtroom: reflections on a decade in the culture of pain. PMID- 11901776 TI - Compassionate utilitarianism: the unknown Bentham revealed. PMID- 11901777 TI - Pitfalls in detaining the 'dangerous'. PMID- 11901778 TI - Assessing psychiatric problems in ethnic minority patients. PMID- 11901779 TI - Reducing repeated deliberate self-harm. PMID- 11901781 TI - Treating obsessive compulsive disorder. PMID- 11901780 TI - Managing perinatal psychiatric problems. PMID- 11901782 TI - Practical diagnosis and management of seizures. PMID- 11901783 TI - Getting ready for the oral. PMID- 11901785 TI - Outcomes associated with restored and unrestored deciduous molar teeth. AB - INTRODUCTION: Most dental treatment for children in the United Kingdom (UK) is provided by general dental practitioners (GDPs) working in the National Health Service (NHS). A working party of the British Society of Paediatric Dentistry, in a special publication from the Dental Practice Board, has suggested that failure to provide restorative care for the deciduous dentition is unacceptable, yet GDPs are filling fewer teeth in young children. The study aimed to evaluate the health outcomes obtained from restoring carious deciduous molar teeth. METHOD: The dental records of 677 children cared for by 50 GDPs in the north west of England were analysed. RESULTS: The results showed that 18.8% of deciduous molars with unrestored caries and 17.0% with a history of restorative care went on to be extracted because of pain or sepsis. CONCLUSION: The results suggest that the risk of carious deciduous molars being extracted is similar whether these teeth receive restorative care or not. PMID- 11901786 TI - Saving pulps--a biological basis. An overview. AB - AIM: Cavity preparation and restoration variables have an uncertain relationship to pulp injury and repair responses. The purpose of this paper is to determine the importance of cavity preparation and restoration variables, by ranking them according to their effect on pulp injury (odontoblast survival) and pulp repair (reactionary dentine secretion). METHOD: The seven studies reviewed are all based on the protocols described by Murray, Smith and colleagues in 2000 and 2001. RESULTS: The studies reviewed provide new perspectives on the importance of cavity preparation and restoration variables in mediating pulp activity. CONCLUSIONS: The onset of post-operative complications may be most productively minimised by focusing practitioner attention to aspects of treatment highlighted in this review. PMID- 11901787 TI - Cough syrup addiction and rampant caries: a report of two cases. AB - Sugar-containing syrups such as cough medications have been recognised as potentially cariogenic, especially for chronically sick children suffering from otitis media, seizure disorder, congenital cardiac diseases and asthma. Unfortunately, an increasing number of juveniles are addicted to cough syrups containing centrally-acting drugs and these individuals not only end up with medical and social problems, but also with dental manifestations. This paper addresses the latter problem with the help of two illustrative cases. PMID- 11901788 TI - Vocational training today and tomorrow. PMID- 11901789 TI - An audit on the placement and replacement of restorations in a general dental practice. AB - OBJECTIVES: To determine the reasons for placing and replacing restorations in a general dental practice amongst a group of patients. METHODS: This study used the method devised by Mjor and later modified by Burke and others. Instruction and data collection sheets were sent out to general dental practitioners (GDPs) for participation in a multicentre study. The GDPs were asked to record consecutively all restorations placed over a period of one month. The author is presenting his data collected over a period of six months using the same methodology. The author's results will be compared with a VDP study and the multicentre study. RESULTS: 779 restorations were placed during the six months of the study. Three hundred and sixty-four (47%) were initial placements and 415 (53%) replacements. Primary fractured teeth totalled (16%) of the restorations. Other reasons for restoration placement were marginal fracture/degradation (31%), non-carious defects (13%) and both primary and secondary caries (9%). The proportion of glass ionomer restorations increased with the age of the patient compared with amalgam and composites (chi-square: p < 0.001), averaging 64% in age groups between 35 and 75 years, increasing to 88% in those patients over 75 years. CONCLUSIONS: The principal reasons for placement and replacement of restorations in this study were repairs to tooth fractures. The proportion of amalgams and composites decreased with age while the proportion of glass ionomers increased. PMID- 11901790 TI - Advances in orthodontics. AB - There has been tremendous progress in orthodontics since Edward Angle first popularised the fixed orthodontic appliance at the turn of the century. Recent years have seen an increased demand for orthodontic treatment from both adolescents and adults and, in addition, patient and clinician expectations of treatment outcomes continue to rise. A desire for more aesthetic materials has resulted in both smaller and 'tooth-coloured' appliances. Improvements in technology, often outside orthodontics, have also led to the development of new materials. The best example of this was the development of nickel titanium alloy by the NASA space programme, which was subsequently adapted for use in nickel titanium archwires. Other technological advances adopted for use in orthodontics include magnets, computerised imaging systems and distraction osteogenesis. This review paper looks at some of the innovations in the fields of materials as well as in techniques and appliance systems. PMID- 11901791 TI - Introducing problem-based learning to special needs dentistry--a case study. AB - INTRODUCTION: In order to promote training and education in special-needs dentistry an attempt was made to introduce problem-based learning (PBL) as a method of postgraduate dental education. The aim of this paper was to review the principles of PBL and report on a case study using this methodology. METHOD: The case study was of a PBL session, on the subject of 'problems of obtaining appropriate dental care for people with epilepsy', undertaken at a national conference. Delegates were asked to complete a pre- and post-session questionnaire on PBL and their attitudes to the session. RESULTS: The session received a mixed response. Only 33 (35%) thought the session was valuable and only 20 (31%) thought it was better than conventional teaching methods and yet over half (55%) said they would like to attend more PBL in special-needs dentistry. Professionals complementary to dentistry were more likely to find the PBL session of value and to prefer the method to a more conventional format than dentists were (chi-square = 5.5, df = 1, p < 0.05 and chi-square = 5.9, df = 1, p < 0.05 respectively). CONCLUSION: Valuable feedback was received from delegates. This will enable improvements to be made in future courses so that the effectiveness of PBL can be optimised. PMID- 11901792 TI - Molecular diagnostics: a powerful new component of the healthcare value chain. PMID- 11901793 TI - n-CoDeR concept: unique types of antibodies for diagnostic use and therapy. AB - The n-CoDeR recombinant antibody gene libraries are built on a single master framework, into which diverse in vivo-formed complementarity determining regions (CDRs) are allowed to recombine. These CDRs are sampled from in vivo-processed and proof-read gene sequences, thus ensuring an optimal level of correctly folded and functional molecules. By the modularized assembly process, up to six CDRs can be varied at the same time, providing a possibility for the creation of a hitherto undescribed genetic and functional variation. The n-CoDeR antibody gene libraries can be used to select highly specific, human antibody fragments with specificities to virtually any antigen, including carbohydrates and human self proteins and with affinities down into the subnanomolar range. Furthermore, combining CDRs sampled from in vivo-processed sequences into a single framework result in molecules exhibiting a lower immunogenicity compared to normal human immunoglobulins, as determined by computer analyses. The distinguished features of the n-CoDeR libraries in the therapeutic and diagnostic areas are discussed. PMID- 11901794 TI - Microfluidic arrays in genetic analysis. AB - The goal of genetic analysis is to discover genetic markers that are informative for providing high confidence, positive predictive value in managing phenotypic outcomes. Primary consensus sequence data, genetic polymorphism databases and associated phenotype data are rapidly making genetic analysis more useful. Therefore, genetic analysis applications are gradually becoming more mainstream. The diversity and complexity of genetic analysis currently requires an array of analytical techniques, instrument platforms and software to support all the steps from data acquisition to interpretation. As supporting research technologies mature, they are incorporating increasing levels of automation, system integration and miniaturization. Microfluidic arrays are positioned to play a key role in routine genetic analysis, particularly as they begin to appear in more fully integrated analytical platforms. PMID- 11901795 TI - MALDI TOF mass spectrometry: an emerging platform for genomics and diagnostics. AB - Matrix-assisted laser desorption/ionization-time of flight (MALDI TOF) mass spectrometry has become, in recent years, a tool of choice for large molecule analyses. The platform is ideal for analysis of protein and nucleic acid sequence, structure and purity. MALDI TOF is the method of choice for quality assurance in oligo and peptide synthesis. Exact mass measurements along with signal intensity detection provide a high level of quality assurance as to the accuracy of the measurement. This accuracy has the potential to significantly lower the level of indeterminate assays that require retest. PMID- 11901796 TI - PCR-CTPP: a new genotyping technique in the era of genetic epidemiology. AB - A new PCR method, PCR-CTPP (polymerase chain reaction with confronting two-pair primers) was invented to genotype a relatively large number of samples in a cost effective and time-saving manner. In this method, allele-specific DNA products are amplified by means of applying appropriately designed two-pair primers (four primers) into an ordinary PCR tube. Single genotyping for G2886T at L-myc, Arg72Pro of p53 and Glu487Lys of ALDH2 as well as duplex genotyping for C-31T of IL-1B with VNTR of IL-1RN and A385T of secretor gene with se5, are demonstrated as examples with the primers and PCR conditions. PMID- 11901797 TI - New developments in the diagnosis of maternal and congenital CMV infection. AB - Recent advances in the screening of pregnant women with Cytomegalovirus (CMV) IgM, CMV IgG and CMV IgG avidity serologic tests, has led to a more accurate diagnosis of CMV infection. When serologic screening is performed early in gestation, it is possible to identify those women at risk of intrauterine transmission of the virus, i.e., those women with a primary CMV infection, who should be enrolled in prenatal diagnosis. The use of quantitative PCR on amniotic fluid from pregnant women at 21-22 weeks of gestation in prenatal diagnosis is an effective diagnostic tool to distinguish between CMV infection and CMV disease in the fetus and newborn. Quantitative PCR on peripheral blood leukocytes from CMV infected newborns can be used to monitor viral load, especially during treatment with ganciclovir. These advances in serology and quantitative virology should lead to more accurate diagnosis of maternal and congenital CMV infection. PMID- 11901798 TI - DNA array technology and diagnostic microbiology. AB - Near instantaneous detection of pathogens from clinical material, combined with simultaneous prediction of their antimicrobial resistance profiles, would revolutionize the impact of microbiology on the management of infection. Array based assays allow a range of characteristics to be rapidly and simultaneously determined. At present these systems have found their primary role as research tools for the monitoring of mRNA expression in the form of DNA microarrays or 'chips'. As fabrication costs reduce and validated targeted arrays are developed, it is inevitable they will be used for more routine applications. Microfluidics offers the exciting possibility of combining purification, amplification and detection in a single disposable device; microarrays are particularly suitable for use within these systems. Arrays will become an important tool for clinical diagnostics. PMID- 11901799 TI - Detecting recurrent bladder cancer: new methods and biomarkers. AB - In this review, a series of biomarkers and molecular assays are compared with conventional urothelial cytology in their ability to detect recurrent bladder cancer. The tests considered in detail include the BTA test, NMP 22 test, DNA ploidy measurements, telomerase determinations and microsatellite instability assays. Although all of these measurements show some degree of improvement for cancer detection, the microsatellite instability assay shows the highest sensitivity and specificity. Additional biomarkers considered in the review include bladder cancer tumor antigens, growth factors, cell adhesion molecules and various molecular markers including cell cycle regulatory genes and p53 mutations. PMID- 11901800 TI - Molecular profiling of tissue samples using laser capture microdissection. AB - The management of cancer and other genetically based diseases is far from optimal in even our most advanced medical centers. There is still uncertainty regarding how diseases will progress in certain patients, toxicity that must be tolerated with imprecise treatment regimens and significant potential for treatment failure. As our understanding of the complexity of these diseases has increased, it has become clear that we must move toward precisely tailored approaches to treating each individual patient. To that end, a major goal of current medical research is the rapid identification of the specific molecular alterations in each patient's disease. This will enable the design of optimal diagnosis, prognosis and treatment, significantly improving survival. This review describes one important approach to the genetic analysis of disease--molecular profiling- and the tenets and technologies necessary for its success. PMID- 11901801 TI - Molecular typing methods for the epidemiological identification of Clostridium difficile strains. AB - Toxigenic Clostridium difficile is the etiologic agent of C. difficile-associated diarrhea (CDAD), the most common cause of nosocomial diarrhea. Cross-infection between patients and transmission through the environment and medical personnel are important factors in the acquisition of CDAD. In order to understand differences in epidemiology and pathogenesis, a number of typing schemes have been developed. We will review the typing methods used to study the epidemiology of C. difficile infections and how they have evolved from a phenotypic identification to state of the art molecular methods, detecting genetic polymorphisms among strains. These molecular methods include PCR-based methods (arbitrarily primed-PCR [AP-PCR] and PCR ribotyping), restriction endonuclease analysis (REA) and pulse field gel electrophoresis (PFGE). The application, usefulness and feasibility of these methods are compared and discussed. Finally, the role of genomics as a tool to investigate CDAD is introduced. PMID- 11901802 TI - Diagnosis of microsatellite instability-positive colorectal cancer. AB - The continuing increase in knowledge regarding the genetic basis of carcinogenesis has led to diverse efforts to exploit this knowledge clinically, primarily in the form of predictive genetic testing. In conjunction with family history, gene tests are intended to improve individual cancer risk assessment. At present, genetic testing for colorectal cancer (CRC) risk--in the form of microsatellite instability (MSI) screening and DNA sequencing--is applied in hereditary nonpolyposis colorectal cancer (HNPCC). In this inherited colorectal tumor syndrome, determining the genetic status may result in an individually tailored surveillance program and prophylactic treatment, reducing cancer morbidity and mortality. PMID- 11901804 TI - New developments in PCR-based diagnosis. Report of a session at the 11th European Congress of Clinical Microbiology and Infectious Diseases Istanbul, Turkey 1-4 April, 2001. PMID- 11901805 TI - Mutation detection using fluorescent hybridization probes and melting curve analysis. AB - The LightCycler is a real-time PCR instrument that combines a thermocycler and a micro-volume fluorimeter. LightCycler technology is gaining popularity due to its ability to detect mutations quickly and accurately. Multiple base alterations are discriminated using hybridization probes and fluorescent melting curves. This review focuses on mutation detection and base discrimination by fluorescent hybridization probes. Assay designs for single base mutation detection and complex multiplex reactions are discussed. Types of mutations detected and reported applications are reviewed. Guidelines using melting curve analysis for the clinical laboratory are presented. PMID- 11901806 TI - Standardization and quality control in molecular diagnostics. PMID- 11901803 TI - Universal Linkage System: versatile nucleic acid labeling technique. AB - Over the last two decades nonradioactive nucleic acid labeling and detection systems have overcome the safety, disposal, stability and cost problems that are associated with radioactive techniques. Besides traditional, enzyme-mediated, nonradioactive labeling methods (e.g., random priming, nick translation or labeling by PCR), several chemical labeling systems have been developed (e.g., ULS, psoralen, alkylating agents). These methods provide fluorescent (or hapten) labeled probes for fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH) and microarray-based techniques. In this review the DNA-based molecular diagnostic applications and perspectives of the Universal Linkage System (ULS) technology will be described. PMID- 11901807 TI - Genomics 101: what the practicing physician needs to know about the genetics revolution. June 8-9, 2001, Chicago, IL, USA. PMID- 11901808 TI - NAATs to diagnose Chlamydia trachomatis genital infection: a promise still unfulfilled. AB - Nucleic acid amplification tests (NAATs) have created a revolution in our ability to diagnose chlamydial infections. They are markedly more sensitive (while maintaining exquisite specificity) than previously used tests. They can be used with noninvasively collected specimens (first-catch urine for men or women and vaginal swabs from women). This allows their use in screening asymptomatic individuals who represent the bulk of prevalent infections. Mishandling of urine specimens can lead to false-negative results and few are aware of that potential for getting incorrect results. The leading deterrent to the acceptance of NAATs has been their perceived cost. This is based on the purchase price of tests and fails to consider the total cost of testing, which also includes costs of specimen collection, where NAATs have a major advantage. PMID- 11901809 TI - Array-based proteomics: the latest chip challenge. AB - Array-based protein technologies are emerging for basic biological research, molecular diagnostics and therapeutic development with the potential of providing parallel functional analysis of hundreds or perhaps hundreds of thousands of proteins simultaneously. Array-based methods are becoming prevalent within proteomics research due to the desire to analyze proteins in an analogous format to that of the DNA microarray. Novel protein biochips are under development in academic laboratories and emerging biotechnology companies to advance the pace and scope of scientific discovery. This review will define array-based proteomics, its current applications and future directions, as well as examine the challenges and limitations of this projected billion dollar industry. PMID- 11901810 TI - Clinical utility of viral quantification as a tool for disease monitoring. AB - The possibility to detect viral DNA or RNA in a quantitative manner has already contributed significantly to the management and diagnosis of viral infections, as well as to the understanding of virus-host interactions. New developments in amplification techniques based on real-time detection, as well as automation of the whole process, will soon be introduced in a diagnostic laboratory setting, thereby enabling a rapid turnaround time to generate both quantitative and qualitative results. New guidelines for disease management, as well as extensive quality control and standardization programs must be introduced. PMID- 11901811 TI - Detection and diagnosis of mycobacterial pathogens using PCR. AB - In the year 2001, it is estimated that 3 million people will die from tuberculosis, caused by the infectious agent, Mycobacterium tuberculosis. After decades of decline in the disease, the resurgence of tuberculosis seen worldwide in the 1990s sparked a renewed interest and commitment of funds for research into M. tuberculosis and other pathogenic mycobacterial species. The discovery of the PCR in the 1980s has had a major influence on the progress made possible in the study of these fastidious, tough-walled and slow-growing mycobacterial species. In the last 10 years, PCR has allowed us to amplify parts of the genome, decipher the nucleotide sequence, discover new mycobacterial species, determine epidemiological relationships between strains and identify genetic changes involved in drug resistance. PMID- 11901812 TI - Molecular methods for rapid detection of group B streptococci. AB - Group B streptococci (GBS) are an important cause of neonatal sepsis and meningitis. Implementation of selective intrapartum chemoprophylaxis based on either a screening-based approach or a risk-based approach has led to a substantial decrease in the morbidity and mortality of GBS disease. Current 'gold standard' detection methods for GBS are selective broth cultures of combined vaginal and anal specimens collected at 35-37 week's gestation. Rapid immunological detection methods, including latex agglutination test, enzyme immunoassay and optical immunoassay, as well as hybridization-based test, are available. These methods are useful in rapid identification of heavily colonized women, but are unable to detect light GBS colonization due to poor sensitivity. Recent development of real-time PCR and fluorescence labeling technologies has provided new detection platforms for bacterial identification. GBS-specific PCR assays using these new technologies offer promising tools for sensitive and specific detection of GBS directly from clinical specimens. The application of these assays in the current prevention strategy will simplify the prevention practice and rationalize antibiotic use. PMID- 11901813 TI - Human tissue kallikrein gene family: a rich source of novel disease biomarkers. AB - The organization of the human tissue kallikrein gene family has now been fully elucidated. This family contains 15 genes encoding secreted serine proteases, which share significant homologies at both the DNA and amino acid level. Two members of the human kallikrein gene family, prostate-specific antigen and human kallikrein 2, have already found important clinical application as prostate cancer biomarkers. In this review, we examine the diagnostic and prognostic value of the 15 human kallikrein genes and proteins. It is clear that at least a few members show promise of becoming novel cancer and other disease biomarkers. PMID- 11901814 TI - Detection of immunoglobulin heavy chain gene rearrangements in hematologic malignancies. AB - B-cell precursors undergo a unique somatic immunoglobulin heavy chain gene rearrangement process. The generated VHDHJH junction is a successful marker in lymphoproliferative malignancies at initial diagnosis for detection of clonality and during treatment for monitoring minimal residual disease. VHDHJH errors are often involved in recurring structural chromosomal aberrations of lymphoid malignancies, with consequent deregulated expression of the juxtaposed oncogenes, e.g., c-myc, bcl-2 or CCND1. Besides cytogenetics, a variety of molecular techniques are becoming increasingly established, including Southern blotting, PCR and real-time PCR, as well as fluorescence in situ hybridisation. Different approaches may be chosen to evaluate lymphoid malignancies either at diagnosis or follow-up in the light of increasing relevance and proven clinical utility. PMID- 11901816 TI - Biological indicators for the identification of ionizing radiation exposure in humans. AB - While the effects of acute high-dose irradiation are well-documented, less is known about the effects of low level chronic radiation exposure. Physical dosimetry cannot always be relied upon, so dose estimates and determination of past radiation exposure must often be based upon biological indicators. Some of the established methods used in the assessment of nuclear accidents are reviewed here, including cytogenetic analyses, mutation-based assays and electron spin resonance. As interest in research on low-level radiation exposures expands, there is an increasing need for new biomarkers that can identify exposed individuals in human populations. Developments in high-throughput gene expression profiling may enable future development of a rapid and noninvasive testing method for application to potentially exposed populations. PMID- 11901815 TI - Telomerase in human tumors: molecular diagnosis and clinical significance. AB - Shortening of structures known as telomeres, which cap the ends of chromosomes, is postulated to limit the lifespan of human cells. Activation of telomerase, an enzyme that synthesizes telomeric DNA, is an essential step in cell immortalization. Telomerase is ordinarily inactive in most somatic cells, but can be detected in nearly all tumors. The activation of telomerase in malignant cancers seems to be an important step in tumorigenesis, whereby the cell gains the ability of indefinite proliferation. Due to the association between telomerase expression and malignancy, the enzyme is expected to be a useful tumor marker and a new anticancer therapeutic target. However, recent results scale down to some extent the initial enthusiastic expectations for telomerase as the ideal malignancy marker. PMID- 11901818 TI - Diagnostic tests for fragile X syndrome. AB - Fragile X syndrome is a common X-linked hereditary disease, characterized by mental retardation, macroorchidism and mild facial abnormalities and is almost always caused by the absence or deficit of the FMR1 protein. In the majority of cases, the disease is associated with an expansion of a CGG repeat, located in the 5' UTR of the FMR1 gene. Diagnostic methods include PCR amplification and Southern blotting, which are performed on DNA isolated from peripheral leukocytes. Recently, varying immunocytochemical tests have been described to identify fragile X patients, based on the detection of FMR1 protein in cells by a monoclonal antibody. This review provides an update on the different DNA methods and gives specific attention to both the newly developed PCR method and antibody methods for prenatal and postnatal diagnosis of the fragile X syndrome. PMID- 11901817 TI - Molecular genetics of X-linked mental retardation: a complex picture emerging. AB - Mental retardation or intellectual disability is a heterogeneous group of disorders of the human brain affecting 2-3% of the general population. It is becoming evident that a large proportion of mental retardation is genetically determined, which means that it can be molecularly defined and thus precisely diagnosed. Building knowledge and understanding about molecular processes leading to 'malfunction of human brain' will clearly bring benefits to patient management, disease prevention and ultimately disease treatment and will also assist in tackling much harder questions of the molecular basis of human cognitive ability. In this review the current knowledge of the molecular genetics of X-chromosome-linked mental retardation and its nonspecific forms in particular is discussed, together with limitations affecting diagnosis and likely new approaches that need to be implemented. PMID- 11901819 TI - Diagnostic and scientific applications of TaqMan real-time PCR in neuroblastomas. AB - This review summarizes the present data on the diagnostic and scientific applications of TaqMan real-time PCR in neuroblastomas, a novel fluorescence based method for quantitation of gene expression and gene copy number variations as gene amplification and locus haploinsufficiencies. Particular emphasis is spent on precision and accuracy and the comparison of TaqMan real-time PCR with more traditional methods for quantitative PCR. Provided a proper assessment for each new marker, this method has been shown to be an easy, fast and reliable alternative to the more traditional methods for the determination of MYCN amplification and neuropeptide Y gene expression. The combination with single cell picking in the neuroblastoma tissue may be a future target of the application of real-time PCR. PMID- 11901820 TI - Beyond our genome: the application of proteomics in molecular diagnostics. PMID- 11901822 TI - Down syndrome collaboration. PMID- 11901823 TI - IGEN to continue MesoScale Diagnostics joint venture. PMID- 11901821 TI - Orchid BioSciences launches ELUCIGENE CF29 screening test. PMID- 11901824 TI - Genesis Bioventures to acquire Biomedical Diagnostics. PMID- 11901825 TI - Xanthon and Chiron to collaborate. PMID- 11901826 TI - Studies confirm clinical utility of Ventana's INFORM HPV ASR. PMID- 11901827 TI - Microscopic cantilever aids assay for prostate cancer. PMID- 11901828 TI - Genetic diagnostics: harnessing technologies to transform diagnostics. June 28 29, 2001, Venice-Lido, Italy. PMID- 11901829 TI - EuroBiochips 2001. June 6-8 2001, Munich, Germany. PMID- 11901830 TI - Pharmacogenetics: the Rx perspective. AB - Pharmacogenetics is changing the way medicines are discovered, developed and delivered to patients. In this article, we present the 'prescription' perspective -how the results of pharmacogenetic research will help minimize the risk of costly adverse drug reactions and treatment failures, by providing predictive tools to enable healthcare providers to prescribe the right medicine for the right patient. We discuss the challenges of this research and its clinical application; its implications for drug development; evolving concepts of informed consent; related ethical, legal and social issues; changing definitions of 'genetic testing'; and the creation of an international Pharmacogenetics Working Group. PMID- 11901831 TI - Genome- and proteome-based technologies: status and applications in the postgenomic era. AB - Genes are transcribed to pre-mRNA, further processed to various mRNAs and then translated into proteins that may be post-translationally modified and subsequently function as the ultimate effecting molecules in the cell. Diagnostic options may be addressed as hybridization-based techniques to monitor nucleotide mutations or transcript levels. These techniques are highly suitable for high throughput analyses based on DNA chip technology. They will enter the diagnostic practice as routine assays, although some obstacles must be addressed. Proteomics based techniques are less suitable for high-throughput analyses at the moment, but are closer to the functional level. The combination of 2-dimensional polyacrylamide gel electrophoresis and mass spectrometry analyses make a strong couple that may enter as diagnostic applications once more automated. PMID- 11901832 TI - Genetic strategies for the personalization of antipsychotic treatment. AB - Pharmacogenetic research into complex traits, such as response to antipsychotic treatment has proved a difficult task. Nevertheless, investigation of drug metabolic enzymes has revealed polymorphisms in specific cytochrome P450 genes responsible for treatment-induced toxic reactions. However, the picture becomes more complicated when drug target sites are investigated in search of genetic influence. Most antipsychotic drugs are multitarget, denoting a complex mechanism of action. Although individual genes have been reported to influence antipsychotic response, no single gene can account for the variability observed in treatment response. Current investigations focus on single gene variants that may be associated with particular side effects or symptoms as well as contributing to general response. The scope of this article is to review recent advances of pharmacogenetic research on antipsychotic drugs and the strategies under development for the individualization of treatment. PMID- 11901833 TI - Testing for antineutrophil cytoplasmic antibodies. AB - The most common reason to request a test for antineutrophil cytoplasmic antibodies (ANCA) is to diagnose Wegener's granulomatosis and microscopic polyangiitis and to monitor inflammatory activity in these diseases. Several retrospective and prospective studies have suggested that the demonstration of ANCA lacks sensitivity and specificity, but these series have detected ANCA with neutrophil-indirect immunofluorescence alone, have used a disease classification that did not describe microscopic polyangiitis and have included patients with inactive disease. The 'International Consensus Statement on Testing and Reporting ANCA' has been developed to optimize the clinical relevance of ANCA testing by the adoption of standardized testing and reporting procedures. International collaborative efforts continue to focus on improving the tests for ANCA. PMID- 11901834 TI - Detection of Helicobacter pylori virulence-associated genes. AB - Helicobacter pylori is an important human pathogen and persistent colonization of the human gastric mucosa can cause severe gastrointestinal diseases. The bacterium should not be considered as a uniform organism, but as a population of closely related and yet genetically diverse bacteria. Several genes of H. pylori (such as vacA and cagA) have been identified as being virulence-associated and may have important clinical and epidemiological implications. Assessment of virulence-associated genes of H. pylori should be included in clinical and epidemiological studies as well as therapeutic trials, in order to stratify between patient groups, harboring H. pylori strains with particular virulence genotypes. Molecular determination of antibiotic resistance will be especially useful for treatment studies. Together with our increasing knowledge about the human genome, typing of H. pylori will facilitate the management of gastroenterological pathologies. PMID- 11901835 TI - Molecular techniques in Whipple's disease. AB - Whipple's disease is a systemic infection, caused by the bacterium Tropheryma whipplei, with protean clinical manifestations characterized by fever, weight loss, diarrhea, polyarthritis, skin hyperpigmentation and adenopathy. For a long time, due to the inability to culture the causative organism, diagnosis was based on histologic examination of infected tissues, usually duodenal biopsies, which revealed diastase-resistant periodic acid-Schiff-positive staining. Now, PCR of various tissues or fluid is emerging as a way to diagnose Whipple's disease. However, the presence of T. whipplei DNA in saliva, gastric juice or duodenal biopsies of healthy individuals has led to questions regarding the specificity of the molecular techniques involved. After a series of failures, stable culture was achieved in 2000. Subsequently, the generation of rabbit polyclonal antibodies has led to the detection of the bacterium in tissues by immunohistology. However, culture and immunohistology are very recent techniques and are not yet widely used. Propagation of the bacterium will lead to extensive molecular knowledge of T. whipplei, which will help in the diagnosis and understanding of the epidemiology and pathogenicity of Whipple's disease. PMID- 11901836 TI - Hepatitis C virus infection: early diagnosis and identification of response to antiviral therapy. AB - Early diagnosis of hepatitis C infection and early identification of virologic response to antiviral therapy represent major hallmarks of the quality of a case. They contribute to reducing the risk of hepatitis C infection from blood product and improve disease management in patients treated with antivirals. Some of the current issues and perspectives involved in detection and quantification of viral load during the incubation phase of infection and monitoring the early phase of antiviral therapy are discussed. PMID- 11901837 TI - Cytokeratin-derived markers of lung cancer. AB - Various evaluation methods are available for aiding clinicians in lung cancer management. Some of these methods are highly specific. However, they are also invasive and burdened by non-negligible complication rates (e.g., mediastinoscopy); other methods are highly accurate and noninvasive, but require expensive equipment and well-trained personnel (e.g., PET scanning); others are fast, inexpensive and safe. However, their diagnostic yield is low and requires further clinical testing (an example of such tests is the chest-x-ray film). There is probably only one way to perform an easy, inexpensive, repeatable test, which is also fairly accurate and predictive. This is tumor marker testing, which -as a large and specialized literature shows--can be highly effective when based on a cytokeratin-derived marker assay. PMID- 11901838 TI - Endocrine tumors of the digestive tract and pancreas: histogenesis, diagnosis and molecular basis. AB - Although relatively rare, endocrine tumors of the digestive tract and pancreas have been widely investigated and represent a complex tumor entity. The two major categories of well-differentiated and poorly differentiated tumors show important phenotypic and clinical differences. In well-differentiated tumors the multiple endocrine neoplasia syndrome of Type 1 (MEN1) gene is frequently abnormal, though a complex multiple gene involvement is postulated for different tumor types. Poorly differentiated carcinomas show frequent p53 gene hyperexpression/defects, characterizing severe cell abnormality and possibly accounting for the malignancy of such carcinomas. PMID- 11901840 TI - PD-loop technology: PNA openers at work. AB - A concise survey of the emerging PD-loop technology is presented, which outlines several exemplary methods with robust DNA diagnostic potential: duplex DNA capture, topological DNA labeling, nondenaturing DNA sequencing and hybridization of molecular beacons to double-stranded DNA. Advantages of these new PNA-based assays over existing techniques for sequence-specific detection and manipulation of DNA duplexes are discussed. Future prospects for the further development of PD loop technology are highlighted. PMID- 11901839 TI - Association mapping: where we've been, where we're going. AB - This paper provides a review of recent work in the area of marker-phenotype association studies, specifically as used for localizing--or mapping--genes affecting a trait of interest. We describe the basis of association mapping and discuss a number of the commonly used techniques. We have also included references to various papers that have evaluated the use of these methods. PMID- 11901841 TI - Towards high-throughput genotyping of SNPs by dynamic allele-specific hybridization. AB - Analysis of single nucleotide polymorphisms (SNPs)--the most common form of variation in the human genome--has become a popular strategy for discovering genes involved in complex diseases such as Alzheimer's disease, obesity and diabetes. It is also widely anticipated that SNPs will play a major role in pharmacogenomics, where the identification of variations in specific genes relevant to drug efficacy, toxicity and metabolism will help to establish optimal therapeutic strategies for individual patients. Reflecting these expectations, many new SNP-related technologies have appeared over the past few years, each with unique advantages, but all with the common goal of simplifying and expediting SNP analysis. We recently introduced a technique termed dynamic allele specific hybridization (DASH), a convenient method for SNP (and insertion deletion) genotyping, which is highly applicable to both basic research and clinical diagnostics. Commercial DASH devices are now available, making the technology affordably accessible for all laboratories. PMID- 11901842 TI - Arrays, molecular diagnostics, personalized therapy and informatics. PMID- 11901844 TI - Pyrosequencing establishes two new collaborations. PMID- 11901843 TI - Molecular diagnostic markets must prove effectiveness. PMID- 11901845 TI - Potential screening test for prostate cancer. PMID- 11901846 TI - Abbott Laboratories to acquire Vysis. PMID- 11901847 TI - ACLARA and Roche diagnostics collaborate. PMID- 11901848 TI - Cholestech and Bayer diagnostics in comarketing agreement. PMID- 11901849 TI - diaDexus grants license to Quest Diagnostics for osteoporosis diagnostic test. PMID- 11901850 TI - SYN-X pharma introduces Alzheimer's disease diagnostic aid. PMID- 11901851 TI - Pharmacogenetics: the Dx perspective. AB - The promise of the rapidly developing field of pharmacogenetics is that genetically determined propensities of individual patients to respond favorably or adversely to a given pharmacologic agent will be able to be determined prior to administration of that drug. The realization of that promise, however, is predicated on a number of developments in the capabilities of diagnostic laboratories. These developments include the introduction of automated technologies for efficiently and accurately detecting, quantifying and decoding specific nucleic acid sequences and the concomitant availability of information technology-based applications for rapidly analyzing, interpreting and then communicating complex genetic data to healthcare providers. This article will review currently available and developmental molecular diagnostic technology and in addition, describe the current status and speculate on the future of pharmacogenetic testing in the clinical laboratory. PMID- 11901852 TI - Genome medicine promised by microarray technology. AB - The human genome project has now been completed, which markedly changes the way to analyze gene functions. Recently developed DNA microarray technologies enable us to explore genome-wide gene expression in the diseased tissues. In this review, we introduce the principles and applications of microarray technologies (such as DNA, tissue and cell microarrays) to molecular diagnostics, drug target discovery and validation of drug effects. PMID- 11901853 TI - Microsatellites: perspectives and potentials of mass spectrometric analysis. AB - Mass spectrometry is a powerful analytical tool in biotechnology. The 'soft' ionization and desorption technologies matrix-assisted laser desorption/ionization and electrospray ionization have enabled mass spectrometric analysis of large biomolecules, such as proteins and nucleic acid amplification products, and paved the way for mass spectrometry to become a leading technology in current genomics and proteomics efforts. Large-scale analysis of single nucleotide polymorphisms by mass spectrometry has been commercially established. This article reviews applications of mass spectrometry for microsatellite analysis. Features and capabilities of the two most prominent techniques, matrix assisted laser desorption/ionization and electrospray-ionization mass spectrometry, are compared and their potential to address the limitations of conventional microsatellite analysis based on comparison of gel electrophoretic mobilities is explored. PMID- 11901854 TI - Molecular detection and genotyping of human papillomavirus. AB - Human papillomavirus infections are associated with the development of cervical neoplasia. Human papillomavirus is a group of heterogeneous viruses, comprising many genotypes, which can be divided into high-risk and low-risk types, depending on their association with disease. Therefore, accurate molecular diagnostic tools are required for detection and identification of human papillomavirus. Monitoring of human papillomavirus infection is necessary for adequate patient management and follow-up during treatment. This review describes the different molecular methods available for human papillomavirus detection and identification of genotypes. PMID- 11901855 TI - Molecular diagnostics in monogenic and multifactorial forms of type 2 diabetes. AB - Type 2 diabetes represents a major and increasing contributor to morbidity and mortality worldwide. Of the world's population, 10% either have Type 2 diabetes, or will develop it during their lifetime. Realization that inherited factors play a leading role in determining individual susceptibility to this condition provides a framework for improved molecular understanding of disease pathogenesis through susceptibility gene discovery. In particular, identification and characterization of these susceptibility variants should lead to improved targeting of available preventative and therapeutic measures and increasingly 'personalized' therapy. This article surveys present knowledge about the genetic basis of Type 2 diabetes and discusses the current and future role of genetic diagnostics, with an emphasis on lessons learned from study of monogenic forms of the disease. PMID- 11901856 TI - Recent advances in the diagnosis of Lyme disease. AB - Diagnosis of human Lyme borreliosis is usually based on serology, which has a number of pitfalls. In the early phase of the disease serology can still be negative, whereas false-positive results are also common. The interpretation of confirmatory Western blot tests is not always easy. Furthermore, routine serology cannot discriminate between active and past infection. In addition, recombinant antigens are being introduced to improve serologic tests. New developments in the diagnosis of Lyme disease are the development of PCR tests. This review gives an overview of the molecular diagnostic possibilities of Lyme borreliosis, mainly by PCR, and describes some interesting possibilities for future serology. PMID- 11901857 TI - Thymidine kinase: diagnostic and prognostic potential. AB - Thymidine kinase is a cell cycle-dependent marker that can be detected in the serum of patients diagnosed with many different types of cancer. Serum levels of thymidine kinase have also been shown to reflect the progression of cancer as well as an indication of the efficacy of chemotherapeutic intervention. A new monoclonal antibody assay for thymidine kinase has been developed, which is capable of detecting thymidine kinase in both serum and tumor tissue. Thymidine kinase assay kits should be available at low cost and could serve as an effective low cost test for the detection and progression of many types of human cancer. PMID- 11901858 TI - Molecular diagnosis of animal diseases: some experiences over the past decade. AB - The experiences of a veterinary research and diagnostic laboratory are summarized on the development and application of the PCR to diagnose a wide range of viral diseases in animals. The group started the routine diagnostic application of the PCR as early as 1988 and today a total of 35 nested PCR assays are in routine use for the detection of 15 DNA and 20 RNA viruses. Special tools and laboratory practice were applied to avoid false-positive results, while false-negatives are avoided by internal controls (mimics). At present, the classical nested PCR methods are being replaced by real-time TaqMan and molecular beacon assays and the multiplex real-time PCR detection of viruses is also under development. By direct sequencing of the PCR products, phylogeny studies are performed and molecular epizootiology results are provided for rapid and exact identification of virus variants. Molecular epizootiology also contributes to trace the routes of virus spreading on large geographic areas. Recently, large efforts have been made to follow the recommendations of Office International des Epizooties for the standardization and international harmonization of the molecular diagnostic assays. PMID- 11901859 TI - Molecular diagnostic testing for infectious diseases using TMA technology. AB - Molecular diagnostic tests based on nucleic acid amplification technologies (NAATs) have become widely established in clinical microbiology laboratories in recent years. The acceptance of these tests has been driven by the development of more accurate and less labor-intensive commercial assay kits by diagnostic manufacturers. Infectious disease diagnostic assays using transcription-mediated amplification (TMA) NAAT have become increasingly popular in many clinical microbiology laboratories. Recent technology developments have improved the performance and simplified the use of the TMA assays. These new technologies have been applied to the development of multiplex TMA tests to improve the testing accuracy for organisms, such as Chlamydia trachomatis and Neisseria gonorrhoeae in clinical microbiology laboratories. TMA tests for HIV-1 and HCV have also led to improvements in blood bank testing which can improve the safety of the public blood supply. PMID- 11901860 TI - Nanobeads as a solid phase in a solute homogeneous assay format. AB - The measurement of numerous samples as in drug screening or diagnostics has been improved significantly over recent years. The processing of a great number of carriers with 96, 384 or 1536 wells is not the limiting step anymore and more than 100,000 samples can be analyzed within 24 h. New challenges arise in optimizing data quality and assay volume in order to gain more reliable results with a reduced consumption of assay reagents. Both are addressed with an approach using dispersed nanoparticles analyzed by a new detection technology, fluorescence intensity distribution analysis. This homogeneous assay is a generic format holding great potential in expanding the assay strategy portfolio for diagnostic and screening applications. PMID- 11901883 TI - [Prognostic value of the assessment of ischemic status at admission to the coronary care unit after pre-hospital thrombolysis]. AB - The object of this study was to assess and analyse TIMI (Thrombolysis in Myocardial Infarction) grade and secondary major cardiac events of patients with acute myocardial infarction benefiting from pre-hospital thrombolysis according to their predefined clinical and electrical "ischaemic status" (Active, Inactive, Intermediate) on admission to the coronary care unit and at the end of thrombolysis (90th minute). This single centre study was undertaken from March 1994 to August 1999 on 161 patients treated by thrombolysis by the emergency ambulance service for acute myocardial infarction (< or = 6 hours). The mean age was 56.2 +/- 11.3 years with 8.7% of women. On admission to the coronary care unit. 30.8% were classified as Inactive and 51.6% as Active. At the end of thrombolysis, 62.3% were classified as Inactive and 27.7% as Active. Nearly 93% of TIMI 3 flow was observed in Inactive patients at the 90th minute and 76.7% of TIMI < or = 2 flow in Active patients (p < 0.0001). Global hospital mortality was 2.48% but it was zero in the Inactive group at the end of thrombolysis. With an average follow-up of 26.9 +/- 15.8 months, the incidence of major cardiac events was 34.1%, including 16.1% of revascularisation of the target lesion. In multivariate analysis, predictive factors for TIMI < or = 2 at the end of thrombolysis included persistent pain, the number of leads with ST elevation and the absence of regression of ST elevation on admission to the coronary care unit. The only predictive factor for secondary major cardiac events was persistent ST elevation at the 90th minute of thrombolysis. PMID- 11901884 TI - [Knowledge and perception of cardiovascular risk factors in Africa South of the Sahara]. AB - The objectives of our study were to evaluate the perception and the knowledge of cardiovascular risk factors (CVRF) by the population and their management by health care workers (HCW) in Burkina Faso. The survey targeted specific socio professional groups representative of Burkina Faso. The survey team administrated a questionnaire and measured some constants. The sampling method was empiric selection by quotas. The sample was composed of 2,000 subjects: 1,073 men, 927 women, 1,800 represents of the general population and 200 HCW. Of 1,800 non health workers, 1,475 had ever heard about hypertension. The representation of hypertension as "a disease" decreased with instruction level while its reprentation "a risk factor" increased with instruction level (p < 0.001). The main sources of information on CVRF were talks with parents and friends, radio broadcasting and discussion with HCW. One hundred and forty of 200 HCW defined hypertension as an elevation of blood pressure, mainly systolic (130 cases) according to WHO criteria. Hypertension was classified after alcohol, tobacco smoking, obesity as the fourth CVRF. One hundred and seventy seven of 302 cases of hypertension were previously unknown: 97 of the 125 old cases were treated and 74 had not normal blood pressure levels. Hypertension and other CVRF are not well known in Burkina Faso population and are not well-managed by HCW. Political decision makers and donor institutions should pay more attention on the public health problem represented by hypertension and other CVRF in developing countries because of they are not communicable. PMID- 11901885 TI - [Contribution of the implantable ECG monitor in the etiologic diagnosis of syncope and unexplained recurrent syncopal attacks. Initial experience with 32 patients]. AB - Following an exhaustive aetiological investigation, 10 to 26% of syncopal attacks remain unexplained. In these cases the correlation between symptoms and rhythm is a deciding step for the aetiological diagnosis. We report our initial experience using an implantable electrocardiographic monitor, a new diagnostic tool in patients suffering from syncope and recurrent unexplained syncopal attacks. RESULTS: The study included 32 patients (average age 55 +/- 22 years; 23 males) suffering from syncope and/or recurrent syncopal attacks remaining unexplained following an exhaustive aetiological investigation. The average follow up was 10.2 +/- 2.5 months. No case of sudden death was registered, and the device was removed in only one patient due to poor tolerance. During follow up, 21 recordings were memorized and analysed in 15 patients (45%), giving an average of 1.4 recordings per patient. The average interval for recurrence of symptoms after implantation was from 84 +/- 104 days, 75% of the episodes coming in the first 2 months following implantation. An arrhythmia was detected on 10 occasions: a malignant ventricular arrhythmia in 2 patients, a non-sustained ventricular tachycardia in 1 patient, a junctional tachycardia in 1 patient, entry into paroxysmal atrial fibrillation in 4 patients, a sinus bradycardia in 1 patient, and a sinus pause for 19 seconds in 1 patient. In one patient ST segment depression was documented following anterior chest pain. The tracing was normal with sinus rhythm recorded on 10 occasions, representing the only documented information in 4 patients. In total, an aetiology was found in 11 of the 32 patients evaluated (34%). Once the aetiological diagnosis was established and a specific treatment initiated, all the patients became asymptomatic. CONCLUSIONS: Our preliminary results underline the significance of the implantable ECG monitor in the diagnostic approach to recurrent unexplained syncopal attacks. The exact place of this tool in the decisional algorithm for syncope remains to be defined with further studies. PMID- 11901886 TI - [Very long-term outcome of 68 vena cava filters percutaneously implanted]. AB - Between January 1987 and December 1991, 68 consecutive patients aged 71.5 +/- 12.0 years underwent percutaneous implantation of a vena caval filter, mainly the LGM (N = 64). Fifty seven patients had pulmonary embolism, 61 had deep vein thrombosis of the lower limbs. The average follow-up interval was 4.9 +/- 3.3 years (7.0 +/- 2.7 years for the patients still alive). The follow-up included a telephonic enquiry to determine the date and cause of death, recurrent deep vein thrombosis and/or pulmonary embolism; surviving patients underwent clinical examination, plain abdominal X-ray with a lateral decubitus view and duplex ultrasonography of the lower limb veins to assess the patency of the filter. Fifty three per cent of the patients died. Four predictive factors for mortality were identified: a contra-indication to anticoagulant therapy, chronic post embolic cor pulmonale, an indication of prophylactic implantation in the elderly and the presence of underlying malignant disease. There were 5.8% recurrences of pulmonary embolism, 26.1% of lower limb deep vein thrombosis and 25% of filter thrombosis. The only predictive factor of thrombosis was a proximal venous thrombus and was associated in 50% of filter thrombosis. Seventy per cent of the plain abdominal X-rays were abnormal with 9 displacements. 9 migrations and 10 closures of the filters. There was a significant correlation between closure on plain abdominal X-ray and caval thrombosis and between recurrent deep vein thrombosis and caval thrombosis. The frequency of long-term complications after implantation of a caval filter in this study suggests that interruption of the vena cava should be reserved for the only validated indications in the presence of a formal contra-indication to or failure of anticoagulant therapy. Other indications require evaluation with prospective randomised trials. PMID- 11901887 TI - [Anticoagulation strategy in coronary angioplasty]. AB - The explosion in the number of angioplasties rapidly imposed a necessary compromise between effective anticoagulation during the procedure to prevent thrombotic phenomena, related mainly to the use of foreign intravascular materials, and the limitation of haemorrhagic complications, the corollary to this type of treatment. Modern universally accepted protocols, the fruit of over 20 years' experience, are described by the authors. However, the constant progress in the management of acute coronary syndromes and the increasing use of new anticoagulant and antiaggregant molecules imply an obligatory adaptation of the use of heparin in all its forms, which should be based on irrefutable scientific evidence: the preliminary results are then discussed. Finally, future trends will be outlined with the development of new, better targeted sites of anticoagulation which may provide optimal safety in the medium term. PMID- 11901888 TI - [Let's make people know our research better!]. PMID- 11901890 TI - [Role of electric stimulation in apnea syndromes]. AB - The sleep apnoea syndrome is the best known apnoeic syndrome. It is observed in 4% of men and 2% of women. Nasal ventilation with continuous positive pressure is the best treatment for most patients. To date, electrical stimulation has a limited role in its treatment as it is used only when the apnoea requires ventilation by tracheotomy. This electrogenic ventilation requires so-called diaphragmatic stimulators. Although severe bradycardia may occur during sleep apnoea, there is usually no indication for cardiac pacing. However, recent publications have reported an anti-apnoeic effect of permanent atrial pacing. The modes of action remain unclear but these results support other recently reported data concerning the value of pacing in cardiac failure, the high incidence of sleep apnoea in cardiac failure patients and the possibility of diagnosing and monitoring apnoea by minute ventilation sensors. Therefore, there appears to be a field of research for cardiac pacing in apnoea syndromes. The authors review the principal reported data on the indications and possibilities of extra-cardiac and cardiac stimulation in apnoeic syndromes. PMID- 11901889 TI - [Stress ultrasonography under low doses of dobutamine, contractile reserve and dilated cardiomyopathy]. AB - The demonstration of a myocardial contractile reserve with low dose dobutamine is an emerging imaging technique in patients with dilated cardiomyopathy. This contractile reserve is correlated with a better prognosis and enables identification of subgroups of patients who could increase their left ventricular ejection fractions under carvedilol. A review of the published literature shows that the method does not expose patients to major risk, providing patients are selected and carefully monitored during the procedure. Complementary studies of larger numbers of patients are required to confirm its value as a prognostic and therapeutic marker in patients with dilated cardiomyopathy. PMID- 11901891 TI - [Primary cardiac lymphoma. Report of a case]. AB - Malignant non-Hodgkins lymphomas have a secondary cardiac localisation in 20% of cases. However, a cardiac primary site is rare (44 cases described up to now). A positive diagnosis is rarely made before death. There is great interest in echocardiography, a non-invasive method, to identify these tumours early. The prognosis remains nevertheless gloomy. We report the case of a child aged 8 years, admitted with a scenario of low output right cardiac insufficiency. Chest radiography identified cardiomegaly with a prominent right border, and the electrocardiograph showed right auricular hypertrophy. A tumour mass infiltrating the right atrium, the right ventricle and the lateral face of the left ventricle was discovered on trans-thoracic echocardiography. Investigation for tumour spread was negative. The patient died before operation in a state of extreme low output. The histology favoured a highly malignant non-Hodgkins lymphoma type B. PMID- 11901893 TI - [The best of 2001. Heart failure]. AB - This year the prevalence, prognosis and the cost of left ventricular function have been clarified. The significance of B natiuretic peptide has been confirmed. The influence of race on the sensitivity to different therapeutic measures has been brought up. Tobacco and diabetes appear as risk factors in cardiac insufficiency. The importance of parietal thickening regarding hypertrophic cardiomyopathy is beginning to be debated, and the risk associated with a new pregnancy in women affected by dilated cardiomyopathy of the peripartum period has been demonstrated. The significance of betablockers is confirmed, although that of angiotensin II receptor antagonists remains a source of questions, despite several advances. Endothelin receptor antagonists present divergent results. Ventricular resynchronisation advances, but its beneficial role regarding the reduction of morbidity and mortality remains to be proved. PMID- 11901894 TI - [The best of 2001. Arrhythmia]. AB - Among the classic problems of rhythmology, which can be classified topographically into atrial rhythm disturbances, junctional rhythm disturbances and ventricular rhythm disturbances, it is certainly the atrial rhythm disorders, including atrial fibrillation, which constitute the "final frontier". Population aging comes with an indisputable increase in the incidence of this arrhythmia, which poses not only classic therapeutic problems (is it better to correct the fibrillation or to be content with slowing it) but also new ones (pulmonary vein tissue ablation for example). It is still very difficult to make a choice among the quantity of ideas and new techniques proposed in rhythmology. From the publications appearing in 2001 in the New England Journal of Medicine, the leading clinical journal, we contemplate topical issues this year concerning three principle subjects: the treatment of atrial fibrillation yet again, arrhythmias of genetic origin, and ventricular rhythm disturbances in the era of the implantable defibrillator. PMID- 11901892 TI - [Immediate and long-term results of radiofrequency ablation of accessory atrioventricular pathways]. AB - The aim of this study was to report the authors' experience of radiofrequency ablation of accessory atrioventricular pathways over a 10 year period (01-91 to 10-00), and the effect of the "learning curve" on the results. The data of 400 patients admitted to primo-ablation of a bundle of Kent was analysed retrospectively. A total of 481 ablations were performed (1.20 per patient). The cumulative global success in the 414 accessory pathways treated was 90.6%. The primary success rate increased from the 1st to the 4th quartile from 68 to 97% (p = 0.0001). The mean duration of fluoroscopy and number of ablation sites decreased from the 1st to the 4th quartile respectively from 47 +/- 27 to 25 +/- 18 minutes (p = 0.0001) and from 8.5 +/- 7.8 to 4.5 +/- 3.8 minutes (p = 0.0001). The average recurrence rate over the four quartiles was 3.6. The overall complication rate was 1.44%. The improved primary success rate from 1991 to 2000 and, in parallel, the reduction of the number of inappropriate ablation sites and fluoroscopy duration are explained not only by the "learning curve" of our centre but also by the benefits of the application of scientific acquisitions (unipolar recordings, criteria for ablation site localisation...) and technical progress (ablation with temperature monitoring...) over this period. PMID- 11901895 TI - [The best of 2001. Echocardiography]. AB - The year 2001 will be characterised by the confirmation of the essential place of harmonic imaging in echocardiography, a technique which has improved in a very important fashion the definition of the endocardial contour, thus giving a more reliable analysis of the segment kinetics, as much at rest as during a physical or pharmacological stress. The advances in the field of stress echocardiography essentially give confirmation of the excellent prognostic value of this technique, either post-infarction or in chronic coronary disease, including hypertensive patients, and with left ventricular hypertrophy or a conduction disorder. The aspirations of a more quantitative approach to regional myocardial function with the aid of myocardial tissue Doppler have been partly confirmed, even if no simple parameter can yet provide information of the presence of myocardial segment viability or ischaemia. The contribution of Doppler echocardiography in the unraveling of left ventricular diastolic insufficiency is crucial, with the description of parameters allowing characterisation of the type of left ventricular diastolic function anomaly and non-invasive evaluation of the filling pressures. The advances in transoesophageal echocardiography have essentially provided a better characterisation of the thrombo-embolic risk markers in atrial fibrillation and in the search for a cardiac source of embolus. Valuable information has equally been provided in acute aortic pathology particularly in the description of symptomatology and evolution of isthmic ruptures of the aorta. The first images of transthoracic echocardiography with real time three-dimensional reconstruction have finally been presented, allowing a glimpse of the enormous possibilities of this method, compared to recorded reconstruction which is routinely performed, especially in transoesophageal echocardiography. Though myocardial contrast echocardiography continues to induce very many fundamental publications, the clinical applications are slow to convince, facing the difficulties of proposing an adequate treatment and standardising the image, and an interpretation which takes account of physiological parameters (coronary micro-circulation). PMID- 11901896 TI - [The best of 2001. Heart stimulation]. AB - Despite the profusion of articles reported in 2001 related to cardiac stimulation, only 5 will be analysed in this review. The choice which has guided this arbitrary selection is the significance of the results for all cardiologists and not exclusively for a group of specialists. Cardiac stimulation continues advancing little by little in the treatment of severe cardiac insufficiency in patients with a complete left branch block. This year the MIRACLE study takes the limelight. Its very clear protocol (stimulation in one group compared to no stimulation in the other) reaches a conclusion which can be considered as indisputable given the large number of patients included: biventricular stimulation significantly improves the functional state of patients at 6 months. In the framework of incapacitating reflex syncope a third randomised study on highly selected patients confirms the indication for this special therapy, which is cardiac stimulation in this situation, even when it is compared with pharmacological "therapy" (betablockade). Syncope, supplying the greatest indication for stimulation, has indisputably made a "breakthrough" in 2001 with the publication of the first recommendations on the subject by a working group of the European Society of Cardiology and thanks to the publication of 2 studies which used implantable Holters to try to clarify their mechanism and to improve the management of patients. PMID- 11901897 TI - [The best of 2001. Clinical pharmacology]. AB - In the field of cardiovascular pharmacology the year 2001 has been marked by the demonstration of the clinical significance of new anti-thrombotics and by the interesting results with anti-endothelins. Whereas the failure of oral antiGPIIb IIIa has been confirmed, melagatran (an anti-thrombin administered orally) and pentasaccharide (a new subcutaneous anti-Xa) have proved their efficacy in the prevention of venous thromboembolic disease compared to low molecular weight heparins, with an acceptable incidence of unwanted effects. Anti-endothelins under development have variable mechanisms of action from one drug to another. Their efficacy in cardiac insufficiency, pulmonary artery hypertension and arterial hypertension is suggested by clinical studies investigating small population; a more important study in decompensated cardiac insufficiency has not however shown a reduction of the morbidity and mortality with one of these drugs. The clinical significance of angiotensin II receptor antagonists has been confirmed for the indications which they share with ACE inhibitors (cardiac insufficiency, prevention of diabetic nephropathy). However, there are not enough comparative trials for these indications between these two classes in order to draw conclusions about the equivalence or superiority of one or the other. Of help elsewhere has been a re-interpretation of the mode of action of arterial wall drugs, and the inflammatory theory of atherosclerosis, putting the accent for example on the reduction of C reactive protein with a statin, or on the anti inflammatory effect of aspirin. However, one study has not shown any benefit in giving a short course of corticosteroids in unstable angina. A very prominent event in the year 2001 remains the withdrawal of cerivstatin due to fatal rhabdomyolysis. The consequences go far beyond this drug, as its withdrawal justifies a fresh examination of the risk-benefit ratio for all the statins, with the probable corollary of a halt in the escalation of prescriptions. In this context, the new ezetimibe-type cholesterol absorption inhibitors could be a future solution. PMID- 11901898 TI - [The best of 2001. Pediatric cardiology]. AB - During this last year, paediatric cardiology has gone through significant changes, although no capital evolution is to be pointed out. Foetal cardiology, initiated some years ago has been remarkably developed. The rate of cardiac anomalies detected is increasing, which leads to very difficult discussions regarding therapeutic abortion and even increasing legal responsibility as evidenced by "arret Perruche". Interventional cardiology is more and more resorted to and a reliable, safe and efficient procedure which improved atrial septal defect (ASD) occlusion devices. Data on occlusion of ASD and patent foramen ovale confirm the reliability of these methods. In the field of imaging. Nuclear Magnetic Resonance angiography which gadolinium can, in some cases, replace fluoroscopy with clearer pictures. Reinforced interest is given to Primary Pulmonary Hypertension (PPH) due to genetic studies and new therapies. The rate of familial PPH is about 6%. It is an autosomic dominant disorder located on chromosome 2q31-32. New treatment use various prostacyclines administrated by different ways: subcutaneously, orally or by inhalation. It is too early to assess their impact on the prognosis. Finally, we may note the paradoxical importance of congenital heart disease in adult as an outcome of interventional cardiology and follow-up of operated children becoming adults. As the beginning of the 21st century, the definition of paediatric cardiology has considerably evolved. PMID- 11901900 TI - [The best of 2001. Thrombosis]. AB - Although it is impossible to recall all the new developments of the year 2001 regarding thrombosis, we have selected certain results: several, with although "negative" results, are highly interesting because they contradict preliminary studies which could have predicted therapeutic perspectives that would considerably alter the management of a common and serious illness. In this sense, they remind us that the large randomised studies (RCT) remain the necessary and indispensable tool for validating a large scale therapy: others, because they come within the logic of certain earlier results, and in this sense it is good for a clinician to see that a certain logic still exists in cardiology; others again, because they define a certainly real benefit, but whose limits of applicability could sometimes escape clinicians. PMID- 11901899 TI - [The best of 2001. Coronary angioplasty or post-interventional cardiology]. AB - RAVEL, a major advance, represents the cardinal event of the year, if not the last ten years. For the first time zero-rate restenosis has been obtained by the placing of a stent coated with a drug with cytostatic properties. This trial puts the first steps of focal chemotherapy delivered by an endoprosthesis into a clinical form. This spectacular result begs to be confirmed in the long term, as well as in the numerous angiographic forms of stenosis currently indicating the placing of a stent. Such a result in the preventive treatment of restenosis of course casts a shadow on the curative intrastent treatment of restenosis, and especially on intra-coronary radiation. In the areas of acute coronary syndromes, and more particularly of myocardial infarction, the trials of pharmacological interventions have multiplied, bringing scattered information which everyone will find of relevance, and which by the wide choice that they suggest, give the prescriber a real degree of freedom. CAPTIM reveals that the clinical results of pre-hospital lysis followed by angioplasty (usually pre-planned) are identical to those of a primary angioplasty. This is good news for the numerous patients who are victims of an MI far from a centre equipped with interventional facilities. These studies, which do not summarize all the latest interventional work, testify to the evolution of the discipline which, currently controlling the area surrounding angiography of stenosis, is directed towards the focal and systemic pharmacological approach to the lesion. PMID- 11901901 TI - [The best of 2001. Vascular medicine]. AB - Length of treatment after venous thromboembolism remains a controversial issue. Two studies published in 2001 did not demonstrate a higher benefit of long-term treatment compared to short-term treatment. New antithrombotic drugs, such as pentasaccharid and ximelagatran are beginning to appear in this field. Long-term travel as a risk factor for venous thrombolysis has also been studied and a recent study demonstrates that it is a risk factor of pulmonary embolism for a travel above 5000 km. No major advances occurred in the arterial field, except the absence of major differences between carotid surgery and carotid angioplasty in the CAVATAS study. This study required to continue to compare these two therapeutic options in randomized trials. PMID- 11901902 TI - [The best of 2001. Nuclear cardiology and MRI]. AB - The year 2001 has been very fertile for innovations and developments in the field of cardiac imaging. In Nuclear Cardiology, this year has been marked above all by a very large number of studies on the gated-SPECT, which allows joint analysis of the perfusion and contraction of the left ventricle. The contribution of this technique is recognised as being very significant in all fields using conventional tomoscintigraphy (screening for cardiac disease, evaluations of prognosis and viability,...). Innovation has also been focused in the field of radio-tracers, with studies on the very promising markers of apoptosis or cellular hypoxia. Lastly, much work has been conducted on the gamma-camera, especially to adapt it for imaging with the tracers used in positron emission tomography (PET). This will enable the distribution to all the Nuclear Medicine services of FDG imaging, which is the reference technique for studying myocardial viability. In MRI much work has been carried out on sequences using gadolinium, a vascular contrast agent, in order to identify and localise the sequelae of infarction, even when they are limited to the sub-endocardium. The most spectacular technical innovations were probably in the methods allowing an increase of the recorded signal (SENSE) and in updating the imaging sequences in real time, without apnoea and without ECG synchronisation. But, as every year, significant progress has also been made in MRI angiography of the coronary vessels, and thus in the analysis of myocardial tissue perfusion, with sequences which can be used without the injection of contrast medium. PMID- 11901903 TI - [The best of 2001. Valve diseases]. AB - The publications in 2001 regarding valvulopathies have concerned all sectors of this pathology. Aortic valvulopathies are the object of new work supporting the relationship between aortic sclerosis or stenosis and cardiovascular risk factors. They confirm the analogy between lesions of inflammatory origin observed on calcified valves and atherosclerotic plaques (Mohlner). They find higher rates of serum lipids in the case of valvular replacement for stenosis than for aortic insufficiency albeit in an older population (Novaro). Monin shows the possibility of a better pre-operative prognostic approach for advanced aortic stenoses at low transvalvular gradient with left ventricular dysfunction, for which the post operative results are better when low dose stress echocardiography has shown the existence of a contractile reserve. For the results of aortic surgery with biological prostheses it is widely reported that they behave as homografts (O'Brien), stented heterografts (Puvimanasinghe) or stent-less (Hubaut). A controversy exists on the subject of the degenerative mechanism of bioprostheses between the supporters of the immunological hypothesis (Human) and those of the purely degenerative hypothesis (Mitchell). This controversy is far from being insignificant because the infectious or other risks run by patients with bioprostheses are conceivable with the addition of an immuno-suppressant treatment. Among the mitral valvulopathies, insufficiencies with an ischaemic origin have a harmful effect on the long term prognosis even for medium leaks (Grignoni). As for the method of repairing these ischaemic leaks, consensus has not been reached between the proponents of exclusive revascularisation, plasty or replacement (Mickleborough, Otsuji). The quality of the very long term results for mitral plasty by Carpentier's technique for rheumatic mitral insufficiency (Chauvaud) or non-rheumatic (Braunberger, Mohty) is confirmed, especially for the latter. Its feasibility by a minimally invasive approach is reported (Schroeyers). Anticoagulation for prostheses remains one of the challenges for valvular surgery. The addition of a platelet anti-aggregant is not accepted by all, due to the increased haemorrhagic risk. A meta-analysis of 2,199 operations seems in favour of this addition if the dose is weak (Massel). It's a question of an attitude having become normal practice across the Atlantic, but not in Europe (Englberger). PMID- 11901904 TI - [The best of 2001. Arterial hypertension]. AB - Last year, in 2001, the results of several major clinical trials have been published, concerning hypertensive patients with type 2 diabetes (IRMA, RENAAL and IDNT studies) and patients with previous strokes. Angiotensin II antagonists (irbesartan and losartan) are able to reduce the rate of progression of diabetic nephropathy in hypertensive patients with type 2 diabetes. This preventive effect occurs independently of the stage of renal dysfunction (early stage in IRMA, patent nephropathy in RENAAL and advanced nephropathy in IDNT). The PROGRESS study shows that the decrease in blood pressure, in response to an ACE inhibitor/diuretic bitherapy (perindopril/indapamide), in patients with previous minor stroke or transient ischaemic attack, reduces significantly the risk of recurrent stroke. PMID- 11901905 TI - Epidemiology, etiology, and prevention of lung cancer. AB - Over the past century, lung cancer has gone from an obscure disease to the leading cause of cancer death worldwide. Initially an epidemic disease among men in industrialized nations, lung cancer now has become the leading cancer killer in both sexes in the United States and an increasingly common disease of both sexes in developing countries. Lung cancer incidence largely mirrors smoking prevalence, with a latency period of several decades. Other important risk factors for the development of lung cancer include environmental exposure to tobacco smoke, radon, occupational carcinogens, and pre-existing nonmalignant lung disease. Studies in molecular biology have elucidated the role that genetic factors play in modifying an individual's risk for lung cancer. Although chemopreventive agents may be developed to prevent lung cancer, prevention of smoking initiation and promotion of smoking cessation are currently the best weapons to fight lung cancer. No other malignancy has been shown to have such a strong epidemiologic relation between a preventable behavior and incidence of disease. Despite this knowledge, more than 20% of all Americans smoke, and tobacco use is exploding in developing countries. Based on current and projected smoking patterns, it is anticipated that lung cancer will remain the leading cause of cancer death in the world for decades to come. PMID- 11901906 TI - Staging classification of lung cancer. A critical evaluation. AB - The International System for Staging Lung Cancer has been validated as a prognostic index and questioned regarding the implications of factors that require further study. As technology for evaluating the anatomic extent of disease is increasingly refined, the accuracy of clinical staging is greatly improved and provides a major benefit for individualized treatment selection. Advancing knowledge of the origin and development of lung tumors presents the challenge of appropriate integration of this body of science into clinical practice. PMID- 11901907 TI - Imaging techniques for diagnosis and staging of lung cancer. AB - Noninvasive imaging modalities continue to offer opportunities in the diagnosis and staging of lung cancer. Positron emission tomography with fluorodeoxyglucose imaging in lung cancer represents an important, new, and accurate tool in the armamentarium of the radiologist. Computed tomography scanning, MR imaging, and PET play a significant role in the selection of patients for curative lung cancer surgery, decreasing the number of unnecessary thoracotomies. The imaging modalities also help to direct the choice of appropriate invasive and surgical procedures. PMID- 11901908 TI - Bronchoscopy and needle biopsy techniques for diagnosis and staging of lung cancer. AB - Lung cancer is the leading cause of cancer deaths in the United States. The individual therapeutic approach and prognosis depends on accurate diagnosis and staging. Flexible bronchoscopy (FB) and transthoracic needle biopsy (TNB) are the most widely used techniques for this purpose. This article provides a critical overview of indications, diagnostic yield, and limitations of bronchoscopy and TNB in the diagnosis of lung cancer. PMID- 11901909 TI - Functional evaluation before lung resection. AB - Advances in operative technique and perioperative care have reduced surgical morbidity and mortality considerably after pulmonary resections. Various single and combined parameters of functional operability have been proposed to assess the surgical risk. Patients with normal or only slightly impaired pulmonary function (FEV1 and DLCO > or = 80% predicted) and no cardiovascular risk factors can undergo pulmonary resections up to a pneumonectomy without further investigation. For others, exercise testing, pulmonary split-function studies, or a combination of these methods are recommended. Cardiopulmonary exercise testing, most frequently performed as a symptom-limited test with the measurement of VO2max, assesses the pulmonary and cardiovascular reserves. A VO2max of less than 10 mL/kg/minute generally is considered prohibitive for any resection, a value of greater than 20 mL/kg/minute or greater than 75% predicted normal, safe for major resections. Split-function studies are radionuclide-based estimations of the ppo values of various parameters. The currently used ppo parameters are FEV1-ppo, DLCO-ppo, and VO2max-ppo. Suggested cutoff values for safe resection are: FEV1 ppo and DLCO-ppo 40% or greater than predicted, and V(r)O2max-ppo 35% or greater than predicted, combined with an absolute value of greater than or equal to 10 mL/kg/minute. The lowest acceptable ppo values remain to be confirmed by additional prospective studies. Resections involving not more than one lobe usually lead to an early functional deficit followed by recovery. The permanent loss in pulmonary function is small (approximately 10%) and exercise capacity is reduced only slightly or not at all. Pneumonectomy leads to an early permanent loss of about 33% in pulmonary function and approximately 20% in exercise capacity. Pulmonary function tests alone therefore overestimate the functional loss after lung resection. PMID- 11901910 TI - Therapy for stage I and stage II non-small cell lung cancer. AB - Despite complete resection of what seems to be all evident tumor, one third to three quarters of patients with stages I and II NSCLC ultimately succumb to this neoplasm. Patients who are cured of an original NSCLC or small cell cancer remain at risk for a new primary lung cancer. Although the importance of lifelong surveillance is clear, the extent and timing of optimal follow-up remain undefined. Although clinicians refer to the development after treatment of clinically discernible sites of tumor as "recurrence," it is probably more accurate to consider these foci as "persistence"--that is, the locoregional site was not sterilized by surgery, and the distant implants were present from the outset but undetected. Although data are sparse, induction and improved adjuvant therapy for early NSCLC may be helpful. Much further experience is needed. Further study and application of biologic indicators in addition to TNM staging likely will help identify patients at high risk for surgical failure who may benefit by combination treatment. PMID- 11901911 TI - Building a better therapy for stage IIIA non-small cell lung cancer. AB - What do clinicians know about stage IIIA lung cancer? They know accurate staging is critical and requires wide application of mediastinoscopy. They know that surgery and radiation alone each can cure a small subset of patients, and complete resection is of the utmost importance in surgically treated patients. They know that chemotherapy can increase the number of patients cured when combined with definitive radiation, and concurrent chemoradiotherapy seems superior to sequential. Neoadjuvant chemotherapy also seems to cure more patients than surgery alone, but more data are necessary. Trimodality therapy remains a promising but unproved approach in patients with stage IIIA disease. With the exciting new molecularly targeted agents, trials examining quad-modality therapy are just around the corner. PMID- 11901912 TI - Therapy for stage IIIB and stage IV non-small cell lung cancer. AB - The treatment options for unresectable stage III NSCLC include definitive RT, chemotherapy, combined chemoradiotherapy, or supportive care. Compared with radiation alone or chemotherapy alone, the combination of chemotherapy and standard RT confers a modest survival benefit at the cost of increased toxicity for patients with an excellent performance status. For metastatic disease, combination chemotherapy--in particular, platinum-based regimens--improves symptom control and survival. Newer chemotherapeutic agents with higher response rates and favorable toxicity profiles are improving outcome even for the elderly and debilitated patients and those refractory to first-line chemotherapy. Evolving understanding of the molecular events in tumorigenesis is uncovering a host of promising targets for mechanism-based therapy. Many of these novel target modulators likely will require combination with conventional chemotherapy for optimal results. PMID- 11901913 TI - Management of small cell lung cancer. AB - Small cell lung carcinoma typically presents as a central endobronchial lesion in chronic cigarette smokers with hilar enlargement and disseminated disease. The diagnostic pathology should be reviewed by a pathologist accomplished in reading pulmonary pathology, and, if any doubt exists in the diagnosis, additional special stains or diagnostic material should be obtained. Patients with extensive stage disease should be managed by combination chemotherapy, whereas patients with limited stage disease should be treated with etoposide/cisplatin plus concurrent chest irradiation. The chemotherapy should be administered for 4 to 6 months and then should be discontinued. Prophylactic cranial irradiation should be given to patients who achieve a complete remission. Patients should be retreated with chemotherapy if they develop a relapse of their small cell lung cancer. The patients who are followed in complete remission should be observed carefully for second cancers, and appropriate therapy should be administered if the cancer reappears. PMID- 11901915 TI - Paraneoplastic syndromes associated with bronchogenic carcinoma. AB - Paraneoplastic syndromes associated with lung cancer are diverse in their presentation, pathophysiology, and implications. They can be seen as a diagnostic and therapeutic challenge or as an opportunity to detect an otherwise asymptomatic malignancy. Unraveling the mechanisms that produce these syndromes will lead to insight into tumor biology that will be translated into novel approaches for early detection and therapy. PMID- 11901914 TI - Therapeutic bronchoscopy in lung cancer. Laser therapy, electrocautery, brachytherapy, stents, and photodynamic therapy. AB - Therapeutic bronchoscopic techniques such as LPR, EC, brachytherapy, stents, and PDT are effective tools in the palliation and local control of lung cancer. Palliation of malignant tracheobronchial obstruction by LPR, stents, brachytherapy, PDT, or a combination thereof results in relief of dyspnea, hemoptysis, and postobstructive pneumonia. Importantly, it avoids intubation in patients with respiratory distress and facilitates the weaning of patients from MV. In the exciting field of lung cancer screening and treatment of early lung cancer, PDT, brachytherapy, EC, and LPR may represent treatment alternatives to surgical resection, especially in a select group of patients with high surgical risk or favorable endobronchial lesions. Clinicians await the results of future studies, which will (1) better define the impact of each treatment modality on patient care in terms of cost, survival, and improvement in quality of life, and (2) determine the optimal combination therapy relative to bronchoscopic and conventional treatment for effective palliation and cure of lung cancer. PMID- 11901916 TI - Gene therapy for lung neoplasms. AB - The field of gene therapy is still in its infancy, but significant accomplishments have been achieved. The ability to transfer genes safely and successfully into animals and patients clearly has been established. It is highly likely that in the near future, gene therapy will be shown to have clear therapeutic efficacy in diseases such as the treatment of hemophilia (using adeno associated virus vectors) and the stimulation of angiogenesis in peripheral vascular disease and myocardial ischemia. Although only Phase 1 cancer gene therapy trials for thoracic malignancy have been conducted (usually in patients with large tumor burdens and at submaximal doses), there are some hints of efficacy at higher doses of vector in trials for localized malignancy. The studies reviewed in this article demonstrate the first attempts to use gene therapy vectors for lung cancer and mesothelioma. Although none of the diseases studied was "cured," valuable lessons have been learned from these trials, especially in defining the challenges of relatively inefficient and transient delivery of transgene in vivo. Using this knowledge, the second phase of gene therapy research has begun, with a strong focus on developing improved vector technology. Given the progress so far, there is little doubt that gene therapy will become a key approach for the treatment of thoracic malignancies in the near future. PMID- 11901917 TI - Preventing lung cancer by stopping smoking. AB - Much progress has been made in recent years in treating tobacco dependence. The 2000 USPHS Guideline extends the understanding of effective treatments and encourages clinicians to be more diligent in recognizing tobacco users in the practice and more aggressive in treating every tobacco user. The guideline outlines the potential use of the five first-line medications (bupropion, nicotine patches, gum, nasal spray, and inhaler) and the two second-line medications (nortriptyline and clonidine). The use of these medications can be tailored to meet the patients needs and combinations of these medications can be used when appropriate. More intensive treatment, such as residential treatment, may be needed for more addicted smokers. PMID- 11901919 TI - Computed tomography screening for lung cancer. AB - The development of CT technology reopened the lung cancer screening debate. Computed tomography screening for lung cancer certainly meets all the criteria required for an appropriate screening test. First and perhaps most importantly, the disease for which the screening is being performed should have a significant prevalence in the population being studied and be a significant health risk for those afflicted with it. Lung cancer is the leading cause of cancer death in both men and women, and one of the most lethal of all cancers. PMID- 11901918 TI - Lung cancer evolution to preinvasive management. AB - Cancer screening measures generally are accepted for cervical, breast, and colon cancer. With prostate cancer screening, the practice is used broadly, but the evidence for this approach is not established clearly. The aggregate clinical experience with these screening procedures provides a useful precedent to consider in designing a new approach to lung cancer screening. For instance, formidable logistic issues exist in responsibly evaluating the vast at-risk population for lung cancer. Obvious issues include the needs for cost-economy and precise classification procedures. In the field of cervical cancer screening, recent developments in automated image analysis highlight the potential for improving the precision of diagnosis by moving beyond traditional manual cytomorphologic evaluation. The authors have outlined how the overexpression of hnRNP A2/B1 was associated significantly with the eventual development of lung cancer, as corroborated by several independent studies. In discussing this marker, they reviewed several issues with relevance to the general challenge in using biomarkers as screening tools for lung cancer. The experience with cervical cancer screening informs the development of a cellular diagnostic screening platform for lung cancer. Issues such as optimized cell preparation, reproducibility, and assay precision are fundamental to the success of the platform for lung cancer detection. The recent Institute of Medicine study of health care delivery provides an excellent point of departure in outlining the global infrastructure that will be necessary in the evolution of a prevention oriented lung cancer care system. The power of early detection in saving lives from cancer is reflected in the fact that more people die from a single cancer without any validated screening tool (i.e., lung cancer) than die from the aggregate of the four other major cancers, including breast cancer, colon cancer, prostate cancer, and cervical cancer cases--which all have more established early detection approaches. New technologies may provide an opportunity to engage lung cancer routinely at a fundamentally early stage in its natural history, which may provide an opportunity for clinicians to reconsider what may be the best way to manage lung cancer. Further sustained efforts will be required to define the true value of these new approaches through clinical trials as the specialty moves responsibly to routine preventative care of individuals at risk for lung cancer. PMID- 11901920 TI - Diagnosis of roentgenographically occult lung cancer by sputum cytology. AB - The specialty has the knowledge and technology to change the outcome of lung cancer. Lung cancer, diagnosed in early stages, is as curable as all other cancers. Sputum cytology is the initial step in diagnosing roentgenographically occult lung cancer. Sputum cytology is complementary to CT scanning. Sputum cytology identifies small central lesions, and CT scanning discovers peripheral tiny adenocarcinomas. PMID- 11901921 TI - Pathology of lung cancer. AB - This article reviews current concepts in pathologic classification of lung cancer based on 1999 World Health Organization (WHO)/International Association for the Study of Lung Cancer (IASLC) classification. Preinvasive lesions including squamous dysplasia/carcinoma in situ, atypical adenomatous hyperplasia and diffuse idiopathic pulmonary neuroendocrine cell hyperplasia are discussed in addition to current concepts of bronchioloalveolar carcinoma and neuroendocrine tumors. PMID- 11901922 TI - Molecular biology of lung cancer: clinical implications. AB - This review summarizes the rapidly expanding knowledge of the molecular pathogenesis of lung cancer. It is clear that respiratory epithelial cells require many genetic alterations to become invasive and metastatic cancer. Much more is to be learned, but with modern technology. Clinicians can detect "field cancerized" regions and preneoplastic and malignant cells, therefore offering the opportunity to intercede with biomarker-monitored prevention and early detection efforts. Such molecular screening and detection efforts will likely be coupled to advances in low-dose computed tomographic imaging, positron emission tomography scans, and other imaging modalities. Although this molecular marker approach has great potential, there is not yet a molecular marker validated in large prospective trials that has major independent predictive prognostic value. There is an urgent need for large, adequately powered, carefully designed prospective studies to identify clinically useful new biomarkers. Finally, new therapeutic strategies with genetic manipulation, small molecules, antibodies, vaccines, and, particularly, new drugs targeting specific biologic pathways found to be abnormal in lung provide for future optimism. Researchers need to define their individual value, especially when integrated with standard therapies. PMID- 11901923 TI - Columnar-lined esophagus. Definitions. AB - Recognition of a columnar-lined esophagus requires precise criteria by which to delimit the esophagus and the stomach. Endoscopically recognizable landmarks such as the squamocolumnar junction (SCJ or Z-line) can be used to identify structures at the gastroesophageal junction. Once the SCJ is located proximal to the gastroesophageal junction, a columnar-lined segment of esophagus is visible. If biopsy specimens from the columnar-lined segment show specialized intestinal metaplasia, then the patient has Barrett's esophagus (BE), and the extent of the columnar lining determines if it is short- or long-segment BE. PMID- 11901924 TI - Standard antireflux operations in patients who have Barrett's esophagus. Current results. AB - Several therapeutic options exist for patients who have BE, and treatment should be individualized (Fig. 1). The best option in patients who have a high surgical risk or who reject surgery is lifelong conservative treatment, adjusting the PPI dosage with pH-metric controls. In patients who have a low surgical risk the best option is Nissen fundoplication. Only in cases in which esophageal shortening prevents a tension-free fundoplication from being done is a Collis gastroplasty associated with a fundoplication indicated. Other options may be indicated only in exceptional circumstances: (a) duodenal switch, when, after multiple failures with previous surgery, the approach to the esophagogastric junction is extremely difficult; and (b) esophageal resection, when there is a nondilatable esophageal stenosis and in cases in which the histologic study reveals the presence of high grade dysplasia. Whatever treatment is used, an endoscopic surveillance program is mandatory, since, with the exception of total esophagectomy, no therapeutic option completely eliminates the risk for progression to adenocarcinoma. PMID- 11901925 TI - Results of the Collis-Nissen gastroplasty to control reflux disease in patients who have Barrett's esophagus. AB - Barrett's esophagus is an end-stage gastroesophageal reflux complication with a potential for malignant transformation. This condition probably is involved in esophageal cancer being perceived today as the most rapidly increasing cancer in Western countries. Numerous observations suggest that standard antireflux operations fail over time because of long-term inflammatory and fibrotic changes in the esophageal wall that cause shortening of the esophagus. The addition of esophageal elongation by gastroplasty provides a reliable repair by creation of a tension-free repair, whereas the durable antireflux effects are provided by the total fundoplication around the neoesophagus. The restored LES tone further helps control the mucosal damage and the chronic inflammatory changes. Complete regression of the abnormal mucosa still does not occur, and persistent irritation of that mucosa still entails the risk for progression toward dysplasia. The natural history of the columnar-lined mucosa in BE may be altered by medical or surgical intervention. It is too early to judge in which settings these interventions will be meaningful. PMID- 11901926 TI - Results of laparoscopic antireflux operations in patients who have Barrett's esophagus. AB - In most patients who have Barrett's esophagus and who are undergoing open or laparoscopic antireflux surgery, there is a significant improvement in symptom control that is equivalent to that in patients who have uncomplicated gastroesophageal reflux disease. The requirement for reoperation in patients with Barrett's esophagus may be slightly higher, although in the two laparoscopic series published to date, the rate is still only approximately 6%. How much this will increase with longer follow-up, time alone will tell, but given the good results in approximately 95% of patients operated to date, the authors do not believe that the diagnosis of Barrett's esophagus should be considered a blanket contraindication for laparoscopic antireflux surgery. Clearly, in most patients with Barrett's esophagus, an antireflux operation will not result in regression of Barrett's mucosa. It is still unclear whether antireflux surgery provides any protection against subsequent development of esophageal adenocarcinoma. What is clear, however, is that after antireflux surgery, patients who have Barrett's esophagus are still at risk for developing adenocarcinoma and should remain in surveillance programs. The authors believe that laparoscopic antireflux surgery is a safe and effective approach for the cure of reflux-related symptoms in patients who have Barrett's esophagus. PMID- 11901927 TI - Familial factors in the etiology of gastroesophageal reflux disease, Barrett's esophagus, and esophageal adenocarcinoma. AB - Familial aggregation of gastroesophageal reflux symptoms has been established clearly in patients with gastroesophageal reflux disease and its complications. Preliminary reports of twin studies suggest that this aggregation has a significant genetic component. A genetic predisposition to gastroesophageal reflux may be expressed phenotypically as disordered gastroesophageal motility and hiatus hernia but these disorders may be secondary to chronic gastroesophageal reflux. Linkage studies, the discovery of candidate genes for gastroesophageal reflux, and their phenotypic expression are awaited with interest. PMID- 11901928 TI - Roux-en-Y long limb diversion as the first option for patients who have Barrett's esophagus. AB - In summary, vagotomy plus antrectomy and the Roux-en-Y procedure is based on the following points: (a) patients who have BE show several foregut abnormalities, including incompetent lower esophageal sphincter, impairment in the esophageal clearance, severe gastroesophageal acid reflux, and frequent duodenoesophageal reflux; (b) late results of classic antireflux procedure in BE are poor with a high recurrence rate owing to a progressive loosening of the wrap; (c) the esophageal damage is produced by the injurious component of the refluxate; and (d) among patients who underwent classic antireflux surgery, a certain proportion developed dysplasia or even adenocarcinoma in the follow-up. The authors have observed that the simple correction of the valve is not enough in many cases, because it does not abolish the gastroesophageal reflux but only diminishes it. In patients who have BE and therefore have impaired esophageal clearance, few reflux episodes can maintain or even induce more damage. With the reduction diversion antireflux procedure, the quality of the corrected valve is secondary, and the main goal is to avoid the reflux of injurious components of the refluxate instead of the refluxate itself, which is almost always impossible. Late results support this hypothesis, and the authors propose this surgical procedure as an alternative treatment in patients who have complicated BE or in patients who have long-segment BE. Among patients who have gastroesophageal reflux and intestinal metaplasia of the cardia or with a noncomplicated short-segment BE, laparoscopic antireflux surgery is the authors' first choice, and only the late objective evaluation of surgical treatment demonstrates which surgical technique is the more adequate to a particular patient who has BE. PMID- 11901929 TI - Role of mucosal ablative therapy in the treatment of the columnar-lined esophagus. AB - With the high prevalence of gastroesophageal reflux-like symptoms in the United States and the association between GERD symptoms and the premalignant condition of BE, there is more and more demand for new and efficacious techniques to treat BE. A wide variety of endoscopic mucosal ablative techniques have been developed with promising initial results. Long-term control of neoplastic risk, however, has not been demonstrated, and most studies demonstrate that there is still potentially some intestinal mucosa present underneath squamous mucosa. Currently, more study is needed to determine which patient groups require therapy of any kind and to determine which therapies would be the most efficacious. Genetic markers may aid in identification of subgroups that are at risk for cancer and help to identify those who would respond to mucosal therapy. Even in patients who have HGD, subgroups of patients who have focal HGD have been found to have better prognosis than those who have more widespread HGD. Currently, there is sufficient information to consider mucosal ablative techniques in patients who are not good surgical candidates. Photodynamic therapy, APC, KTP, Nd:YAG and argon lasers, MPEC, and EMR may provide good alternatives, depending on the degree of dysplasia, the extent of disease, and the age of the patient. Photodynamic therapy and Nd:YAG laser therapy have been applied to more neoplastic lesions, whereas KTP:YAG, APC, and multipolar coagulation have been successful in nondysplastic Barrett's mucosa. In the future, there will be more information to justify the application of mucosal ablative therapy in selected patients. PMID- 11901930 TI - Experimental Barrett's esophagus and the origin of intestinal metaplasia. AB - Many questions remain unanswered regarding the pathogenesis and the cell origin of Barrett's esophagus. Recent studies suggest that progenitor cell populations, which are presumed to reside at the basal layer in the squamous epithelium and at the esophageal glands duct epithelium, may differentiate into a glandular phenotype leading to the development of columnar epithelium in the distal esophagus. Other studies also support the hypothesis that cardiac epithelium may precede the occurrence of specialized intestinal metaplasia. It remains unclear whether cardiac-type epithelium in Barrett's esophagus arises from squamous epithelium or from migration of native cardiac epithelium at the EGJ into the distal esophagus. Experimental animal models of chronic reflux esophagitis, although with some shortcomings when researchers extrapolate the study data to the human situation, have provided interesting insights into possible mechanisms associated with the occurrence of Barrett's esophagus. A better understanding of the molecular mechanisms regulating the development of Barrett's esophagus is necessary for developing new strategies directed toward prevention and treatment of this metaplastic condition with a potential risk for malignant transformation. PMID- 11901931 TI - Role of acid and bile in the genesis of Barrett's esophagus. AB - Clinical and experimental studies have shown that acid and bile reflux are increased in patients who have Barrett's esophagus. The combination of both seems the key factor in the pathogenesis of Barrett's esophagus. This factor has been confirmed by immunohistochemical studies that show that environmental factors, such as acid and bile, are involved in the pathogenesis of Barrett's esophagus. There is a critical pH range between 3 and 6 in which bile acids exist in their soluble, un-ionized form; can penetrate cell membranes; and accumulate within mucosal cells. At a lower pH, bile acids are precipitated, and at a higher pH, bile acids exist in their noninjurious ionized form. Thus incomplete gastric acid suppression, as is the case with most medical treatment regimens for gastroesophageal reflux, may in fact predispose to the development of Barrett's esophagus. PMID- 11901933 TI - Dysplasia in barrett's esophagus. New techniques and markers. AB - In this article, the authors describe the histologic features and classification of dysplasia in Barrett's esophagus. The histologic problems in the diagnosis and grading of dysplasia are discussed. Techniques that can serve as aids to the histologic diagnosis of dysplasia, such as flow cytometry and molecular markers, are reviewed. PMID- 11901932 TI - Endoscopy in Barrett's esophagus. Surveillance during reflux management and new advances in the diagnosis and early detection of dysplasia. AB - Given the alarming rise in the incidence of esophageal cancer and the fact that Barrett's esophagus is clearly a precursor to this disease, effective surveillance is desirable. Endoscopic surveillance is recommended by major endoscopic and gastrointestinal societies based on the available data and hypothetic models suggest that the costs of endoscopic surveillance for Barrett's esophagus may be reasonable when compared with other commonly applied cancer screening strategies. Although, however, most implicated physicians agree that surveillance is warranted, recommended guidelines often are not followed. This occurrence may reflect the importance of some of the practical limitations inherent to carrying out intensive endoscopic biopsy protocols in large numbers of eligible patients. In an effort to improve the surveillance process, several new techniques have been tested and are in development. These techniques are aimed at facilitating the histologic sampling of larger areas of metaplastic epithelium, at better targeting sites more likely to harbor dysplasia and cancer, and at replacing endoscopic biopsies with nonhistologic tissue analysis. Although many of these newer techniques are promising, however, none are currently close to widespread clinical application. The current standard for surveillance remains the use of systematic endoscopic biopsies, with the frequency of surveillance endoscopies determined by the severity of any dysplastic changes that are found. Given the large number of patients that are likely to be eligible for screening and the current constraints in terms of physician availability and health-care resources, endoscopic biopsy will remain the cornerstone of Barrett's esophagus surveillance strategies unless newer alternatives are clearly advantageous in terms of accuracy, cost, availability, and ease of application. In the future, however, advances in techniques for minimally invasive ablation of Barrett's epithelium may make endoscopic surveillance obsolete altogether. PMID- 11901934 TI - Appearance and prognosis of dysplasia in Barrett's esophagus. AB - Dysplasia in a patient who has Barrett's esophagus (BE) is the first step in the neoplastic process. The grade of dysplasia correlates with the likelihood of developing adenocarcinoma of the esophagus during follow-up surveillance endoscopy and biopsy. Surveillance intervals are proposed based on the existing data of the likelihood of developing adenocarcinoma. In the future these intervals will be refined as a result of better definition of the risk factors the development of adenocarcinoma of the esophagus. Risk stratification of the individual patients by age, gender, grade of dysplasia, length of BE, length of hiatal hernia, and presence of genetic mutations will be possible. PMID- 11901935 TI - High-grade dysplasia in Barrett's esophagus. The case for esophagectomy. AB - The main principles for optimal management of HGD arising in Barrett's esophagus are that unequivocal diagnosis of HGD is a prerequisite for making the decision of any kind of treatment. HGD must be resected because of the presence of neoplastic cells in the lamina propria in 40% of patients. No reliable endoscopic or endosonographic feature exists that allows accurate prediction of the existence of neoplastic cells within the lamina propria of a patient having HGD in endoscopic biopsy material. Prompt decision to remove an HGD lesion as soon as unequivocal histologic diagnosis has been settled prevents the development of extraesophageal neoplastic spread. Esophagectomy is preferable to endoscopic mucosal excision because approximately 20% of patients who have HGD in preoperative biopsy material carry neoplastic cells beyond the muscularis mucosae. Esophagectomy can be limited to the removal of the esophageal tube without extended lymphadenectomy because 96% of patients who have HGD in endoscopic biopsy samples have a neoplastic process confined to the esophageal wall. Esophageal resection must encompass all the Barrett's area because of the risk for the further development of a second cancer in the metaplastic remnant. Vagus-sparing esophagectomy with colon interposition or elevation of the antrally innervated stomach up to the neck is preferable to conventional esophagectomy with gastric pull up because the former procedure maintains gastric function intact, whereas the latter exposes patients to the risk for the long-term development of reflux esophagitis and even of metaplastic transformation of the proximal esophageal remnant. Subtle details in the understanding of a given patient's clinical course may be critical for making the decision of the most relevant mode of therapy; therefore, patients who have HGD should be treated in dedicated centers, the experience of which offers the best chances of uneventful recovery if the surgical option is retained. PMID- 11901936 TI - Role of molecular biology in the follow-up of patients who have Barrett's esophagus. AB - At present, the follow-up of patients who have Barrett's esophagus (BE) should occur within the setting of an endoscopic biopsy surveillance program and with the frequency of surveillance as proposed by the American College of Gastroenterology. In the future, patients who have BE will be further stratified according to their risk for progression to invasive carcinoma. This stratification will permit the development of more rational surveillance programs. Models that incorporate epidemiologic risk factors, reflux symptoms, and endoscopic and histologic findings will likely include panels of biomarkers for further stratification of patients as low, intermediate, or high risk. Therefore, the challenge over the next decade will be to define the role of molecular markers in endoscopic surveillance strategies and to identify additional clinically relevant molecular markers for prognosis as intermediate markers for chemoprevention and as molecular targets for novel gene therapies. PMID- 11901937 TI - [The neglected control of proteinuria during pregnancy]. AB - In two women, primigravidae aged 29 and 27 years, no dipstick test for proteinuria was carried out despite symptoms of preeclampsia at 34 5/7 and 25 5/7 weeks of pregnancy, respectively. Both babies died in utero. The women were treated in the intensive care unit; the first woman died due to 'haemolysis, elevated liver enzymes, low platelet count' (HELLP) syndrome, while the second woman was able to return home in a reasonable condition on antihypertensive medication. Dipstick tests for proteinuria should always be carried out in pregnant women with symptoms of preeclampsia in order to avoid the death and serious morbidity which can be associated with eclampsia. PMID- 11901938 TI - [Is there an indication for additional local irradiation in conserving treatment of breast cancer patients aged 60 and over?]. AB - Recent results from the European Organisation for Research and Treatment of Cancer (EORTC) trial of additional irradiation in patients with breast cancer, show that after breast-conserving surgery and radiotherapy (50 Gy) of the whole breast, an additional dose of 16 Gy on the tumour bed significantly reduces the local recurrence rate from 7.3% to 4.3%. A relative reduction was seen in all age groups but was most significant in patients aged 40 years and below (19.5% versus 10.2%). In women aged 60 years and over, the local recurrence rate after radiotherapy of 50 Gy (without the additional radiation dose) is already very low (4.0%). Therefore it is questionable whether an additional dose of 16 Gy (reducing the recurrence rate to 2.5%) is still justified as a standard treatment in this age group. PMID- 11901939 TI - [Reduced risk of stroke recurrence due to hypotensive medication, irrespective of the initial blood pressure]. AB - The 'Perindopril protection against recurrent stroke study' (PROGRESS) demonstrated that for patients with a history of stroke or transient ischaemic attack during the previous 5 years, a blood-pressure-lowering regimen based on the combination of a diuretic and an angiotensin-converting enzyme (ACE) inhibitor, reduces the risk of stroke recurrence (fatal and non-fatal) by 28% (95%-CI: 17-38). This effect was irrespective of the initial neurological diagnosis (ischaemic or haemorrhagic stroke) and blood pressure level. Patients who were treated with just the ACE inhibitor did not exhibit these effects. This large-scale clinical trial demonstrates that hypotensive medication in the form of a diuretic combined with an ACE inhibitor is a beneficial strategy for the secondary prevention of stroke in normotensive and hypertensive patients. PMID- 11901940 TI - [Constitutional eczema; the possibilities of local treatment]. AB - Atopic dermatitis is a common skin disease with a major impact on the quality of life. The cause of this disease is unknown and the diagnosis is made using a set of diagnostic features. In very moderate cases topical treatment of the dry skin may be sufficient. In more severe cases topical treatment with corticosteroids is recommended. Dependent on the steroid used and the severity of the eczema, different application schemes and treatment methods may be used. In exceptional cases, tar derivatives, topical antibacterial compounds and non-steroidal anti inflammatory drugs may be applied. In the near future, the limited therapeutic arsenal of topically effective substances will almost certainly be extended to include drugs such as tacrolimus and pimecrolimus with an inhibiting effect on inflammatory cytokines. PMID- 11901942 TI - [Diagnostic image (78). A young girl with attacks of anxiety]. AB - Asthmatic attacks in a 1.5-year-old girl were followed by expulsion of an Ascaris lumbricoides specimen: the respiratory symptoms were caused by Loeffler's syndrome. PMID- 11901941 TI - [Clinical thinking and decision making in daily practice. An elderly man with hyponatremia]. AB - An 88-year-old man presented with nausea and vomiting. Recently a cutaneous B cell lymphoma had been diagnosed on his right cheek. Laboratory investigation showed hyponatraemia. Fluid restriction was started, based on the diagnosis of the syndrome of inappropriate antidiuretic hormone secretion (SIADH). However the hyponatraemia persisted and a diagnosis of 'reset osmostat' was made. CT of the abdomen revealed slight bilateral adrenal enlargement, which was interpreted as adrenal incidentaloma. No other localisation of the lymphoma, besides that on the right cheek, was seen. Although the symptoms initially disappeared, they recurred and were quickly followed by hypotension. The patient died. Post-mortem examination showed bilateral destruction of the adrenal glands due to lymphoma. The correct diagnosis was Addison's disease. This case shows that diseases do not always present with all the classical symptoms, and that it is important to consider test characteristics of diagnostic tests and to judge investigations in the context of the other clinical findings. PMID- 11901943 TI - [Tissue donation in nursing homes; the situation two years after the introduction of the Organ Donation Act]. AB - OBJECTIVE: To ascertain the state of affairs in Dutch nursing homes with regard to the Organ Donation Act, two years after its introduction. DESIGN: Descriptive, questionnaire study. METHOD: By means of a structured questionnaire the availability, implementation and use of the protocol on tissue donation as well as difficulties in the use of this protocol were enquired about in April 2000 in all 253 Dutch combined and categorical somatic nursing homes. The attitude of nursing home physicians to tissue donation in nursing homes was also assessed. The questionnaire was sent to heads of the Medical Departments, who supplied the answers in cooperation with the nursing home physicians. Finally, partly based on registration data from the Donor Register and from Bio Implant Services, an inventory was made of the number of occasions the national Donor Register was consulted and of the number of tissue donations from nursing homes. RESULTS: In total, 186 questionnaires were returned (response: 73.5%). A protocol on tissue donation had been introduced by 136 (73%) of the Dutch nursing homes, and 96 (52%) of the nursing homes actually used such a protocol. Since the introduction of the Organ Donation Act, the number of tissue donations from nursing homes has not increased: 1-2 skin and 15-20 cornea donors per year. The nursing home physicians were of the opinion that compliance with the protocol required a considerable amount of time, work and emotions, yet due to the large number of contraindications for donation, these efforts were scarcely rewarded. PMID- 11901944 TI - [Malaise after a trip to Africa: amoebic liver abscess]. AB - During a flight to Kenya, a 42-year-old man took a therapeutic dose of chloroquine because of fever. He regularly travelled to Africa and always took chloroquine and proguanil for malaria prophylaxis. The fever disappeared but he did not fully recover. He complained of malaise and weight loss. Fourteen years previously he had suffered from amoebic dysentery. One month after the onset of the patient's complaints, an amoebic liver abscess was suspected on the basis of his medical history, an elevated ESR and a slight leukocytosis. The diagnosis was confirmed by ultrasonography and serology. PMID- 11901945 TI - [Airway fire, a serious complication during laryngeal laser surgery]. AB - During laryngeal laser surgery a 74-year-old male experienced endotracheal tube cuff ignition. This caused severe damage to the trachea. Eventually the patient died after 26 days on the intensive care unit due to a underlying cause. Microlaryngeal and tracheobronchial surgery require a good level of cooperation between the anaesthesiologist and the ENT surgeon, especially when a laser is used. To reduce the risk of an airway fire occurring, a number of precautions can (and must) be taken. Completing a checklist before the laser is used can prove helpful in this respect. PMID- 11901946 TI - [Clinical thinking and decision-making in practice. 4 times ERCP, 6 times echography of the upper abdomen and 3 CT scans in a woman with recurrent fever and bacteremia]. PMID- 11901947 TI - [CBO guideline 'high blood pressure' (revision)]. PMID- 11901948 TI - [CBO guideline 'high blood pressure' (revision)]. PMID- 11901949 TI - ['Caring always'; physicians' expression of condolences to the relatives of their patients]. PMID- 11901950 TI - [Myotoxicity and rhabdomyolisis due to statins]. PMID- 11901951 TI - [The law on corpse care]. PMID- 11901952 TI - ['Doctor, I don't want to know'--investigation of denial in patients with lung cancer]. PMID- 11901953 TI - [Mistakes in methodology. XXXVI. Likelihood ratios and Bayes' rule]. PMID- 11901954 TI - [Diagnostic image (70). A man with fever, swollen joints and erythema nodosum]. PMID- 11901955 TI - [PEG-catheter: a trap without escape?]. PMID- 11901956 TI - The Journal has a long tradition of scientific excellence and rigour. PMID- 11901957 TI - Does size rescaling require central attention? AB - The goal of the present experiment was to determine if object size rescaling in the shape-matching task requires central resources. Two polygons were presented side by side as the second task in a psychological refractory period paradigm. The polygons could be the same as each other or mirror images of each other and the size ratio between the two polygons was varied. In the first task, subjects were required to make a speeded response to the pitch of a tone. The polygon task followed at varying SOAs and the subjects then made a speeded same/mirror image judgement on the polygons. The size ratio effect was additive with SOA indicating that size rescaling is capacity demanding and that it requires central attention. PMID- 11901958 TI - The influence of cue-task association and location on switch cost and alternating switch cost. AB - Task-switching performance is strongly influenced by whether the imperative stimulus uniquely specifies which task to perform: Switch cost is substantial with bivalent stimuli but is greatly reduced with univalent stimuli, suggesting that available contextual information influences processing in task-switching situations. The present study examined whether task-relevant information provided by task cues influences the magnitude of switch cost in a parallel manner. Cues presented 500 ms prior to a trivalent stimulus indicated which of three tasks to perform. These cues either had a preexisting association with the to-be-performed task (verbal cues), or a recently learned association with the task (spatial and shape cues). The results paralleled the effects of stimulus bivalence: substantial switch cost with recently learned cue-task associations and greatly reduced switch cost with preexisting cue-task associations. This suggests that both stimulus-based and cue-based information can activate the relevant task set, possibly providing external support to endogenous control processes. Alternating switch cost, a greater cost for switching back to a recently abandoned task, was also observed with both preexisting and recently learned cue-task associations, but only when all tasks were presented in a consistent spatial location. When spatial location was used to cue the to-be-performed tasks, no alternating-switch cost was observed, suggesting that different processes may be involved when tasks are uniquely located in space. Specification of the nature of these processes may prove to be complex, as post-hoc inspection of the data suggested that for the spatial cue condition, the alternating-switch cost may oscillate between cost and benefit, depending on the relevant task. PMID- 11901959 TI - Letter encoding in visual word identification: more evidence for a word integration explanation for parafoveal preview effects in reading. AB - A letter string presented briefly in the parafovea facilitates naming a foveally presented word provided that the two stimuli are orthographically similar. The facilitation (called priming) is asymmetrical in that to obtain it, both letter strings must have the first letters in common. One possible explanation, a letter integration hypothesis, proposes that readers only identify the letters at the beginning of the parafoveal stimulus, an action that facilitates processing the target. Another explanation, a word-integration hypothesis, postulates that all the letters of the parafoveal stimulus are identified and that the asymmetry occurs because the first letters of the parafoveal stimulus are weighted more heavily than the later ones. The two accounts differ in the way the position of the first letter is determined: The first postulates that readers know the side to identify first without reference to the stimulus; the second postulates that readers establish an order on the stimuli postcategorically. To distinguish the views, we presented English and Hebrew stimuli to bilingual readers. Readers could not anticipate the position of the first letters; hence, if the letter integration explanation is correct, the asymmetry in the priming should be attenuated. Consistent with the word-integration explanation, however, priming occurred when the target shared the beginning letters with the prime in both languages. PMID- 11901960 TI - Strategies of text retrieval: a criterion shift account. AB - This study scrutinized people's ability to apply different strategies to randomly intermixed immediate and delayed test items. In three experiments, participants first read one set of stories. Later, they read more stories, and after each one, answered intermixed questions about that story and one of the earlier ones. The experiments cumulatively manipulated amount of delay, test probe plausibility, probe relation (explicit, paraphrase, inference), and testing procedure (mixed versus uniform delay). Using signal detection response criterion as the index of strategy, we contrasted the single criterion hypothesis, according to which one text retrieval criterion is applied to all test items, and a multiple-criterion hypothesis. The results consistently favoured the multiple-criterion hypothesis. The results also indicated that the presence of immediate and delayed probes mutually influence one another: Less extreme signal detection criteria were adopted under mixed than uniform testing. It was concluded that text retrieval strategy is continually calibrated with reference to the quality of the test probes. PMID- 11901961 TI - Probability matching: encouraging optimal responding in humans. AB - Two hundred undergraduate students participated in a repeated-trials binary choice procedure in which choice of one outcome was correct on 75% of trials. Subjects received 192 trials and were divided into five conditions: (1) control; (2) subjects were given the actual probabilities; (3) subjects were told if they did well they could leave early; (4) competition condition; (5) midway through the task subjects were asked to recommend a strategy for another subject. Half of the subjects in each group were told that the best they could do was to be correct on 75% of the trials. This manipulation permitted assessment of the hypothesis that subjects in probability-matching tasks are seeking a strategy that will be correct on 100% of the trials. The results partially confirmed this hypothesis. In addition, two of the variables improved performance significantly (giving probabilities and asking subjects to recommend a strategy). However, while subjects in all groups improved significantly over trials, optimal choice did not occur in this task. PMID- 11901962 TI - Factors associated with distress in urban residential fire survivors. AB - PURPOSE: To identify factors associated with recovery in a sample of urban residential fire survivors. DESIGN AND METHODS: 440 survivors, of residential fires were interviewed at approximately 3, 6, and 13 months after the fire to measure psychological distress. A set of factors was identified that correlated with survivors' ability to recover from the fire event. Potential predictors of increased distress were identified. Hypotheses were that participants who were lower in socioeconomic status, who were minority members, who had less social support, who engaged in attributional thinking, and had greater concurrent life stresses would have greater psychological distress in response to a residential fire and would be less able to recover from the fire event. FINDINGS: Distress after fire was high at 3 months and decreased for the majority of participants, although one-third of survivors had higher distress at 13 months than at 3 months. Loss of control and attributional variables had the strongest influence on psychological distress over time. CONCLUSIONS: The findings are consistent with stress-response tendencies expected after a stressful event. A set of predictor variables was identified to help clinicians target survivors at high risk for psychological distress after a residential fire. PMID- 11901963 TI - Risk factors for breast cancer in Jordanian women. AB - PURPOSE: To investigate risk factors associated with breast cancer in Jordanian women. DESIGN: Retrospective case-control study based on data from the Jordanian Cancer Registry, in 1996. METHODS: One hundred women with breast cancer (cases) and 100 women without breast cancer (controls) were interviewed in their homes. A questionnaire was developed in Arabic to investigate the risk factors associated with breast cancer in Jordanian women. FINDINGS: Bivariate analysis indicated significant differences between the cases and controls, including age of menarche and menopause, use of households' pesticides, stressful life events, and direct trauma to the breast. Logistic regression analysis indicated higher odds ratios for breast enlargement, irregular menstruation, use of hair dye, oral contraceptives, and fertility drugs. CONCLUSIONS: Significant differences in correlates of breast cancer were found between the cases and the controls. PMID- 11901964 TI - Expressing health experience through embodied language. AB - PURPOSE: To describe embodied language for Japanese elders who suffered a stroke or cardiac disease within the previous year. Embodied language is the overlap of feeling and temporal word use with blood pressure during descriptions of health experience. DESIGN: Exploratory. METHODS: Blood pressure and word use were recorded simultaneously when 17 cardiac and 20 stroke participants described their health experiences for 4 minutes. Blood pressure was measured using a tonometric monitor and word use was measured using linguistic analysis software. Descriptive and nonparametric statistics were used. FINDINGS: Participants with strokes retained higher blood pressure after talking than did cardiac participants. The two groups showed contrasting relationships between word use and blood pressure, particularly for temporal words. CONCLUSIONS: This collaborative research between Japanese and American colleagues was a step toward deciphering shared values, which are important to understanding health for people who have lived through life-changing illness events. PMID- 11901965 TI - Validation of the smoking self-efficacy survey for Taiwanese children. AB - PURPOSE: To determine the psychometric properties of the Chinese version of the smoking self-efficacy (SSE) survey. DESIGN AND METHODS: The SSE survey was translated into Chinese then was back-translated into English, reviewed for content validity, pilot tested, and administered to 401 children between December 1998 and August 1999. A random cluster sampling method was used in this study. FINDINGS: Reliability was indicated by Cronbach's alpha coefficient, .98. The validity of the SSE scale was determined by face validity, item-total correlation coefficient, content validity index, and concurrent validity. Principal component analysis was done to determine the construct validity of the SSE scale. The revised SSE scale had three components accounting for 74.3% of the total variance with alpha of .96. The correlation coefficient between the SSE and revised SSE scale was .99. CONCLUSIONS: Findings show that the revised SSE scale is not only parsimonious but also it is as reliable and valid as is the original SSE scale. This translated instrument is appropriate for use in studies of smoking behavior in Taiwanese children aged 11 to 14 years. Further research will be needed to validate the SSE scale with different populations and settings in Taiwan. PMID- 11901966 TI - Acknowledging unexplained fatigue of tired women. AB - PURPOSE: To investigate indicators of fatigue, including depression, sense of powerlessness, and body aches, and to examine differences between symptomatic and asymptomatic women. DESIGN: Descriptive, comparative analysis. METHODS: Investigators evaluated fatigue, depression, sense of powerlessness, and body aches for two groups of women in a small, rural community. Participants in one group (n = 20) reported subjective experiences of fatigue and the other group (n = 20) did not. No participant had a recognizable disease or physiologic alteration that would account for her fatigue. Symptoms in groups were compared using t tests with Bonferroni adjustment. RESULTS: Although the asymptomatic group members were younger, the groups did not differ in ethnicity, mean weight, number of medications taken, or normality of laboratory values. Women who reported feeling fatigued also had significantly higher scores on the depression and fatigue subscales of the fatigue instrument and significantly lower scores on the power instrument. For participants reporting fatigue, fatigue correlated with depression and depression negatively correlated with sense of power. Data did not indicate how fatigue and depression, or depression and sense of power, are interrelated. CONCLUSIONS: Findings provide support for the importance of acknowledging fatigued women's often readily dismissed complaints not only of fatigue, but of depression and sense of powerlessness, and for conducting further research regarding these complaints in women with no objective indicators of fatigue. PMID- 11901967 TI - Maximizing retention in community-based clinical trials. AB - PURPOSE: To identify and discuss retention strategies and their effectiveness in community-based clinical trials in the last decade. METHODS: Online and hand searches for reports for the period 1990-1999. A total of 87 reports on 64 different trials were found; 21 of these published reports included a description of both retention strategies and outcomes. These studies, rank-ordered on participant retention, were compared. FINDINGS: Despite differences in study populations, interventions, and endpoints, community-based trials with the highest retention rates included a combination of retention strategies. CONCLUSIONS: As more funding agencies emphasize clinical trials, more investigators will be held accountable for credible and generalizable findings based on retaining the projected number of study participants. The small number of published reports for the decade that identified both retention strategies and outcomes substantiates the need to better document these factors in future research reports. PMID- 11901968 TI - A patient-derived perspective of health-related quality of life with peripheral arterial disease. AB - PURPOSE: To evaluate the effects of peripheral artery disease (PAD) on health related quality of life (HRQoL) from the patient's perspective to establish a foundation for systematic PAD-specific HRQoL assessment in this population. DESIGN: Grounded theory methodology. METHODS: Open-ended, tape-recorded interviews were conducted with 38 patients (24 men, 14 women) 44 to 83 years old (mean 65) from two US medical centers to report patient experience of PAD and its perceived effects on HRQoL. Tapes were transcribed and analyzed to identify themes and conceptual domains pertinent to the experience of PAD in this population. RESULTS: Seven major themes were identified: (a) delay in diagnosis and frustration with management of disease; (b) pain; (c) limitation in physical functioning; (d) limitation in social and role functioning; (e) compromise of self; (f) uncertainty and fear; and (g) adaptation to the effects of the disease and demonstration of resiliency. CONCLUSIONS: These findings indicate important psychosocial and emotional consequences of PAD that existing HRQoL questionnaires do not indicate. More complete data might lead to greater understanding of the effects of PAD, serving as the foundation for a more sensitive instrument to assess HRQoL as a basis for more effective interventions. PMID- 11901969 TI - Patients' attitudes toward advance directives. AB - PURPOSE: To explore hospitalized patients' attitudes toward advance directives, their reasons for completing or not completing advance directive forms, and demographic differences between patients who did and did not complete advance directive forms. DESIGN AND METHOD: The convenience sample comprised 30 hospitalized patients in North Carolina. Participants were interviewed using an adapted advance directive attitude survey (ADAS), and were asked five general questions about advance directives. Validity and reliability were established on the adapted tool. FINDINGS: The overwhelming majority of participants had received information on advance directives and they were moderately positive about them. The majority who had completed advance directives were Caucasian, female, over age 65, had less than a high school education, and perceived their health as poor. Most believed that an advance directive would ensure they received the treatment they desired at the end of life. CONCLUSIONS: Patients' attitudes alone did not determine who will and will not complete advance directives. Most participants who completed advance directives had specific reasons for doing so. Nurses have responsibility for discussing advance directives with patients, families, and physicians to ensure adequate education about the completion of advance directives. PMID- 11901970 TI - Does international nurse recruitment influence practice values in U.S. hospitals? AB - PURPOSE: To determine if both U.S. and international nurses place high value on organizational attributes that comprises the elements of professional nursing practice and if the absence of these attributes is associated with high levels of burnout in both U.S. and international nurses. DESIGN: A secondary analysis of survey data was conducted with a sample of 799 nurses; 547 were born in the United States (US) and 252 were from 34 other countries. METHODS: Culture of origin was coded according to Hofstede's classification system. Nursing values related to the professional practice environment were measured using the Nursing Work Index, and burnout was measured using the Maslach Burnout Inventory. Analysis of variance and least-squares regression procedures were used to test hypotheses. FINDINGS: No significant differences were found between U.S. and international nurses in the overall degree to which they valued a professional nursing practice environment. A professional practice environment had a significant, inverse effect on job-related burnout on both U.S. and international nurses. CONCLUSIONS: These findings challenge a prevailing sentiment that international nurses hold values inconsistent with professional practice models as defined in the U.S. context. These findings also show that absence of a professional practice environment will produce similar high levels of burnout in both U.S. and foreign-born nurses. To address the nursing shortage, administrators should ensure that organizational attributes are consistent with a professional nursing practice environment. PMID- 11901972 TI - Patient education for managing pain. PMID- 11901971 TI - Health promotion and participatory action research with South Asian women. AB - PURPOSE: To examine South Asian immigrant women's health promotion issues and to facilitate the creation of emancipatory knowledge and self-understanding regarding health-promoting practices; to promote health education and mobilization for culturally relevant action. METHOD: The study was based on critical social theory; the research model was participatory action research (PAR). Two groups of South Asian women (women from India and of Indian origin) who had immigrated to Canada participated in the project. The qualitative data were generated through focus groups. Reflexive and dialectical critique were used as methods of analyzing qualitative data. The data were interpreted through reiterative process, and dominant themes were identified. FINDINGS: Three themes that were extracted from the data were: (a) the importance of maintaining culture and tradition, (b) placing family needs before self, and (c) surviving by being strong. An issue for action was the risk of intergenerational conflicts leading to alienation of family members. Over a period of 3 years, the following action plans were carried out: (a) workshops for parents and children, (b) sharing of project findings with the community, and (c) a presentation at an annual public health conference. CONCLUSIONS AND IMPLICATIONS: The project activities empowered participants to create and share knowledge, which was then applied toward action for change. Health and health promotion were viewed as functions of the women's relationships to the world around them. PMID- 11901973 TI - Recognizing women's work. PMID- 11901974 TI - Methodological issues in Web-based research. AB - PURPOSE: To examine methodological issues associated with using the Internet and World Wide Web for scientific research, namely, issues related to the nature of the sample, the testing environment and environmental factors, privacy, and confidentiality, and response rates. METHODS: Reviews of literature and personal observation and experience. FINDINGS: Web-based research provides many advantages such as access to specific, sometimes difficult-to-find populations, speed of data access, and decreased costs for data collection and data entry. Such benefits are likely to increase as more nurse scientists use the Web for research purposes. CONCLUSIONS: Nurse researchers who use this technology must consider the methodological problems associated with Web-based methods. The research potential exists, but the methodological issues discussed in this paper are real and, if not addressed, they can seriously affect the validity of study findings. PMID- 11901975 TI - State policies and nurses with substance use disorders. AB - PURPOSE: To investigate the effects of two state regulatory policies on nurses with substance use disorders (SUD) on relapse rates and retention in the nursing workforce. DESIGN & METHODS: This longitudinal comparative study consisted of six data-collection points in 6 months. One sample of 100 registered nurses (RNs) and licensed practical nurses or vocational nurses (LPN/VNs) had disciplinary actions taken against their licenses by four U.S. state regulatory boards using a traditional, disciplinary approach. The alternative sample consisted of 119 RNs and LPN/VNs from three U.S. states where the regulatory board diverted nurses to programs for treatment and for determining suitability to return to practice. Data were collected via mailed surveys. FINDINGS: When compared to the sample, the alternative sample had more nurses with active licenses, fewer with criminal convictions, and more nurses employed in nursing. No difference in relapse rate was found. Fewer than 15% of nurses in either sample experienced one or more episodes of relapse during the 6-month study. Of those who did relapse, the majority used alcohol or street drugs. CONCLUSIONS: Alternative policies worked as well as disciplinary policies and were a more humane and rehabilitative approach. Alternative policy is consistent with the recommendation in Healthy People 2010 to eliminate financial barriers limiting access to treatment. Further investigation is needed to identify situations that trigger relapse. PMID- 11901976 TI - Response of animals to dietary gramine. I. Performance and selected hematological, biochemical and histological parameters in growing chicken, rats and pigs. AB - The effects of feeding varied levels of low- and high-gramine yellow lupin seeds (LG and HG, respectively), and of synthetic gramine added to the diets in amounts ranging from 0.15 to 1.2 g per kg were investigated in one experiment on growing chicken and in two experiments on growing rats. The comparison of LG and HG lupin and the effect of 0.5 g gramine per kg of LG diet were determined in a growth balance experiment with pigs. Organ weights and histology, blood parameters and activity of liver enzymes were determined. The response to HG lupin and gramine concentration varied among the species, the rats being more affected than chicken; no adverse effects of HG lupin or gramine were found in growing pigs. The common reaction of rats and chicken to the high levels of gramine (native or synthetic) was the decrease of feed intake and body gain. The increase of the relative weight of liver or kidney, changes in hematological parameters and liver enzymes were found only in rats. The estimated NOAEL (no-observed-adverse-effect level) of gramine was about 0.3 g/kg diet for rats, 0.65 g for chicken and at least 0.5 g for growing pigs. PMID- 11901977 TI - Response of animals to dietary gramine. II. Effects of feeding high-gramine yellow lupin seeds on reproductive performance of rats and on selected hematological and biochemical parameters in offspring. AB - Two groups of 26 male and 26 female rats at the initial age 30 +/- 2 days were fed during 31 weeks on diets containing 20 percent of yellow lupin seeds having low (LG) or high (HG) gramine content. The animals were mated twice within nutritional groups, 1 male: 1 female, and their main reproductive parameters were recorded. In both reproductive cycles body weight of females at mating, after parturition and after 21-days lactation was lower in HG than in LG group. Fertility rate and body weight of neonates were not affected by the diet while number of neonates per litter tended to be lower by 0.7 and 0.8 pups in HG than in LG group. Body weight of weaners was also substantially smaller in dams fed on HG than LG diet. Relative weight of spleen but not of liver, kidney and heart was significantly greater in HG females. Four weeks old males and females issued from the first litters born to LG and HG animals (ten males and ten females per treatment) were fed individually on respective diets during 3 weeks. Feed intake and growth rate did not differ between the treatments. In males relative weight of liver and testicles was greater, while hematocrit and red blood count were lower in HG than in LG group. In females organ weights did not differ. Activity of liver enzymes determined in males was not affected by the diet. It may be concluded that high-gramine lupin affects negatively lactational performance, probably via lower feed intake, but it does not induce apparent teratogenic effects in the progeny. PMID- 11901978 TI - Effects of mould and toxin contaminated barley on laying hens performance and health. AB - Moulded and mycotoxin containing barley was incorporated into the diets for laying hens to study the effects on performance and health. Health indicators were different blood plasma parameters and liver vitamin A and E levels. A total of 30 hens were fed 3 diets, one supplemented with 30% of toxin-free and two with differently moulded barley from 1997 and 1998 for 7 weeks. The moulded diets contained low to moderate concentrations of ochratoxin A, zearalenone, deoxynivalenol and nivalenol. Inclusion of mouldy barley in the diets had an adverse effect on feed intake, feed conversion, digestibility of nutrients, egg production and egg quality. Plasma alkaline phosphatase was increased and certain biochemical blood parameters (bilirubin, uric acid, chlorine, protein, albumin, vitamin A) were also higher or changed compared to control. The ochratoxin A contamination although relatively low could have contributed to some of these effects as well as reduced intake of feed. The higher mould contamination and an unidentified cell-toxic constituent in the diet containing barley from 1998 can probably also explain the more marked effects from this diet. PMID- 11901979 TI - Influence of high vitamin E dosages on retinol and carotinoid concentration in body tissues and eggs of laying hens. AB - The aim of the study was to contribute to the discussion of overdosing vitamin E in laying hens. A total of 45 laying hens, divided into 5 groups were fed diets supplemented with either 0; 100; 1000; 10,000 or 20,000 mg dl-alpha-tocopheryl acetate/kg diet over a period of 10 weeks. Concentrations of vitamins A and E were measured in plasma, various tissues and egg yolk. Furthermore egg yolk colour and some carotinoids were measured in egg yolks. None of the vitamin E doses significantly influenced performance of the hens. As expected, vitamin E concentration in plasma, all tissue samples and egg yolk was significantly increased with increasing tocopherol content in the diet. The egg yolk showed the highest vitamin E concentration, followed by liver and muscles. Feeding 1000 mg alpha-tocopheryl acetate per kg diet resulted in an increase of vitamin A concentration in the liver. Very high doses (10,000 and 20,000 mg/kg diet) significantly decreased retinol concentration in the liver and egg yolk, as well as carotinoid concentration in the egg yolk. The lower carotinoid concentration in egg yolk resulted in a decreased intensity of egg yolk colour. A prooxidative and/or competitive effect of very high doses of vitamin E with other fat soluble substances has been discussed. PMID- 11901980 TI - Brown and white adipose tissue lipid composition in three successive progenies of rats: effects of ethanol exposure. AB - The effect of ethanol exposure on the fatty acid composition of brown and white adipose tissue in three successive rat progenies at the end of an experimental period (24 weeks) was studied. Ethanol-treated rats received a standard rat chow diet and 5, 10 and 15% ethanol in the ad libitum drinking fluid over 3 successive weeks. Then a concentration of 20% ethanol was maintained for 5 additional weeks up to the end of the experimental period. The males and females in the ethanol treated group were mated to obtain the 1st generation of offspring. Then female and male rats from the 1st generation were mated to obtain the 2nd generation. Finally, males and females from the 2nd generation were mated to obtain the 3rd generation of ethanol treated rats. Another group served as control and received only water and a standard rat chow diet. The control group was handled in the same way as the other experimental groups. In the 1st and 2nd generations the percentage of stearic acid (18:0) decreased and palmitoleic (16:1n7) and oleic acid (18:1n9) increased in both adipose tissues of ethanol-treated rats with respect to control. Additionally, n-3 and n-6 series were reduced both in brown and white adipose tissues. In the 3rd generation the fatty acid composition of the white adipose tissue was similar to that of control rats. Thus, no significant difference in essential fatty acids and oleic acid (18:1n9) were found. However, the fatty acid composition of the brown adipose tissue, in the 3rd generation, was similar to that observed in the 1st and 2nd generation. Thus, a decrease in essential fatty acids and an increase in oleic acid (18:1n9) was found. This suggests adaptation to ethanol consumption during successive progenies in white adipose tissue. However, in brown adipose tissue the values indicate a triglyceride storing during the thermogenesis, which is more important to newborns. PMID- 11901981 TI - Effect of harvest date and variety on ruminal degradability of ensiled maize grains in dairy cows. AB - In this study we investigated the influence of harvest date and genotype on the ruminal degradability of the organic matter of ensiled maize grains. Grains of the varieties Avenir, Byzance, CGS 5104 and CGS 5107 from six different harvest dates were available; they are classified as intermediate types between flint and dent corn. The six harvest dates, during which time the dry matter content of the ensiled grains rose from 52% to 66%, extended from 1st September to 19th October. Assuming a passage rate of k = 0.08, the effective ruminal degradability declined in this period on average from 93% to just under 79%; variety-specific deviations also increased markedly during this period. The dry matter content (x, DM in %) of the ensiled grains had a profound influence on ruminal degradation: a highly significant curvilinear decline in ruminal degradability (y) was calculated at increasing DM levels (k = 0.08), which can be described by the equation y = 0.072x2 (+/- 0.010) + 7.417x (+/- 1.186) - 98.71 (+/- 34.58) (B = 0.96; sy.x[%] = 1.36). The ruminal degradability of ensiled maize grains is about 5-10% higher than that of fresh maize grains. PMID- 11901982 TI - [Cooperation between criminal investigators and forensic medicine. A complementary system and its peculiarities, illustrated by a case from general practice]. AB - Usually criminal investigations involving issues of medical misconduct require comprehensive inquiries which normally include the confiscation of medical files and the determination of certain events in their chronological order. This information is then submitted to the relevant experts as a basis for preparing their opinions. Where the subject of the investigations is complex, however, or where no suitable patient documentations are available, cooperation with the medical experts should start at an earlier stage already and be more comprehensive. On the basis of the relevant legal principles an already concluded investigation ("dietary capsule case") is critically discussed. PMID- 11901983 TI - [An initially unexplained death during prison sentence. Diagnostic verification by atomic absorption spectrometry]. AB - The authors report on the death of a 25-year-old man due to electric current in prison, which was not recognized at first. The fatal accident was caused by a self-made electric kettle. The only electrothermal skin lesions were some pinhead sized electric marks on the left thumb. Careful external inspection of the body in combination with an analysis of the conductor material (aluminium) by means of atomic absorption spectrometry (AAS) revealed the cause of death and the source of accidental electrocution. PMID- 11901985 TI - [Fatal thoracic compression after being partially run over by an automobile]. AB - The authors report on a fatal pedestrian accident in which the victim lay on the street and got under a car being run over by one of the front wheels. Both the car driver and the killed pedestrian were strongly alcohol-intoxicated at the time of the accident (2.45 a.m.). After the car had come to a standstill, the pedestrian remained trapped under the car until she was rescued by the fire brigade. On the basis of the autopsy findings and the technical expert opinion it could be reconstructed that the pedestrian's death was not caused by the consequences of being run over by the car, but by the subsequent compression of her thorax in the final accident position. PMID- 11901984 TI - [An unusual case of 3 fatal, accidental CO poisoning cases by re-routed ventilation]. AB - A case of fatal carbon monoxide poisoning is presented where three members of a family, aged between 23 and 66 years, died while having dinner. After analysing the scene and evaluating the personal circumstances there was no doubt that death was due to accidental carbon monoxide poisoning. The fact that three persons of different ages died almost simultaneously as well as the technical reconstruction appears noteworthy to the authors. Scrutiny of the technical circumstances revealed that the waste air containing carbon monoxide was re-routed because of the oppressively hot weather conditions. The fumes from the stove heating in the living room flowed into the chimney and from there via another pipe into the kitchen oven and finally reached the atmosphere of the kitchen where the three members of the family died. PMID- 11901987 TI - [Snuff. Historical film comments on a current topic]. AB - The movie genre "snuff" appeared in the late 70s and shows the allegedly real, often cruel killing of people. More recently, a growing number of short video clips have been distributed via the internet, which also belong into this category and were clearly recognized as fictional on the basis of technical details by two study groups at the 80th Annual Meeting of the German Society of Legal Medicine held in Interlaken from 25 to 29 September 2001 (Schyma/Seidl). By means of a brief review of film history the article shows that "snuff movies" originated from a certain prevailing trend, examples of which are the murder of the actress Sharon Tate by the group around Charles Manson, the development of B horror movies, a promotional campaign for a film which flopped in 1971 (renamed several times by the distribution company from "American Cannibal" to "Snuff" to "Big Snuff") and elements of urban legends. Retrospectively there are no clues that Snuff films--i.e. movies openly distributed and actually available to the public, which were not private documentations found by the police at the homes of killers during investigations of homicides (such as in the murder series of the couple Bernardo/Homolka)--show real killings. PMID- 11901986 TI - [Mallory-Weiss syndrome as the cause of sudden, unexpected death]. AB - The Mallory-Weiss-syndrome is an upper gastrointestinal haemorrhage due to longitudinal mucosal lacerations in the oesophagogastric junction. The mucosal lacerations occur as a sequel of vomiting or any other increase in intraabdominal pressure and account for up to 15% of all upper gastrointestinal bleedings. If death is due to a bleeding Mallory-Weiss-tear, however, massive blood stains on the corpse and around it frequently give rise to the suspicion of an unnatural cause of death. For this reason, autopsy is usually indispensable to elucidate the circumstances in which death occurred. The authors carried out a retrospective analysis of 5958 autopsies performed between 1997 and 2001 at the Institute of Legal Medicine, University of Hamburg, in order to identify all cases of Mallory-Weiss-syndrome as cause of sudden, unexpected death. The results (9 cases, amounting to 0.15% of all autopsies) suggest that Mallory-Weiss syndrome is probably much more common as a cause of sudden death than previously described. Regarding epidemiological aspects, men clearly predominate; the average age was 48 years. It should also be stressed that in 8 of the 9 cases there was a previous history of chronic alcohol abuse. PMID- 11901988 TI - [Cadaver mix-up--an avoidable problem?]. AB - Depending on local conditions it may happen from time to time that no officer involved in the previous investigation is present during the forensic autopsy. In this case, the post mortem examiner often has to rely solely on written documentation concerning the identity of the deceased. The authors report a case where the body had already been mixed up with another body by the undertaker prior to the inspection by the police. In spite of clear distinguishing characteristics in the written documentation, this led to the wrong body being autopsied. For the post mortem examiner, the only safe way to identify a body seems to be the presence of persons who had known the deceased or of police officers as acknowledging witnesses. If these conditions are not given, other means of safe information on the identity are called for, for example a "police body card" similar to an evidence card. PMID- 11901989 TI - [Theoretical studies of QTL analysis of complex genetic diseases in humans]. AB - Some genes controlling human diseases have been located on the regions less than 1 cM using linkage disequilibrium, and a few of them have been cloned. Z. W. Luo developed a method that can detect and estimate the coefficient of linkage disequilibrium between a marker locus and quantitative trait locus (QTL), and raised the theoretical strategies for high-resolution mapping of complex genetic disorders in humans. Based on the data mentioned above, a method for linkage test between a marker locus and QTL was set up, and a method for linkage test between two marker loci and epistatic QTL was suggested for the first time. PMID- 11901990 TI - [Cloning and sequencing of junction fragment with exon 51 deletion of Dystrophin gene]. AB - To study the mechanism of Dystrophin gene deletion, we obtained the deletion junction fragment of exon 51 by inverse PCR and analyzed the sequence characteristic of breakpoints and deletion junction fragment. After the full sequence of intron 51 was finished, the rough site of breakpoint in intron 51 of a DMD patient with exon 51 deletion was detected by PCR with 9 pairs of primers spaced every 3-5 kb in intron 51. Then the junction fragment was amplified by nested inverse PCR. After sequencing the junction fragment, the 3' breakpoint and partial sequences of intron 50 were determined by comparing with the normal sequences in intron 51. The primer was designated to sequence intron 50 according to the sequence of junction fragment, and then the 5' breakpoint was determined. A total of 1,614 bp in intron 50 was sequenced. The 5' and 3' breakpoints were located in the THE-1 internal sequence (Transposon-like Human Element, THE) and L2 sequence respectively. There are 3 bp junctional homology and no errors near the junction point. This is the second report that the deletion breakpoint located directly in THE-1 sequence studied at the DNA level. We here firstly reported that there is a THE-1 sequence in intron 50. THE-1 and non-homologue end joining repair mechanism may be associated with the Dystrophin gene deletion. PMID- 11901991 TI - [Relationship of growth hormone (GH 2) genotypes with some production performances in pig]. AB - The genotypes of 117 Nanchang White pigs and 361 Large Yorkshire pigs at GH 2 locus were detected by PCR-RFLP. The PCR products were cut by Apa I, and produced two alleles: A(449 + 101 + 55 bp), and B(316 + 133 + 101 + 49 bp). Effects of different genotypes on some important production traits involving the birth weight, 2-month body weight, 6-month body weight, corrected back-fat thickness, average back-fat thickness, feed to gain ratio and lean percentage were analyzed. The results showed that in Nanchang White pigs, no significant differences were observed between different genotypes and different growth and carcass traits; while in Large Yorkshire, the pigs with BB genotype had more lean percentage than pigs with AA genotype (P < 0.05). PMID- 11901992 TI - [Cloning and sequencing of fatty acid binding proteins gene in chicken]. AB - A pair of primers was designed according to the sequence of mammal fatty acid binding protein (FABP) gene, then PCR amplified to chicken genome. After the product of PCR was cloned and sequenced, homologous comparison was done among porcine heart fatty acid binding protein gene and porcine adipocyte fatty acid binding protein gene. The result showed that the sequence of chicken FABP gene had 68% and 75% homology with porcine H-FABP and A-FABP gene respectively, and had 75% homology with porcine AFABP on amino acid level. The result of Northern showed that the gene only expressed in fat tissues. PMID- 11901993 TI - [Studies of single nucleotide polymorphism of PPAR gene and its associations with fattiness trait in chicken]. AB - In this experiment, the AA broiler fat traits and three Chinese special breeds (Shiqiza, Beijing youji and Baier) were used to study the effect of PPAR-alpha gene on fat trait. Coding region of the gene was amplified by seven pairs of primers, and then single nucleotide polymorphisms (SNPs) were detected by the technique of single strand conformation polymorphism (SSCP) and finally confirmed by sequencing. One nucleotide variation was found, the result of chi 2 analysis showed that the distribution of three genotypes was very different among different breeds(P < 0.01). The result of variance analysis showed that the birds with BB genotype had a higher abdominal fat weight than the birds with other genotypes (AA and AB). It implied that PPAR-alpha gene could be a candidate locus or linked to a major gene to significantly affect abdominal fat traits in chicken. PMID- 11901994 TI - [Comparative chromosome painting shows the red panda (Ailurus fulgens) has a highly conserved karyotype]. AB - We have established a comparative chromosome map between red panda (Ailurus fulgens, 2n = 36) and dog by chromosome painting with biotin-labelled chromosome specific probes of the dog. Dog probes specific for the 38 automates delineated 71 homologous segments in the metaphase chromosomes of red panda. Of the 38 autosomal paints, 18 probes each delineated one homologous segment in red panda genome, while the other 20 ones each detected two to five homologous segments. The dog X chromosome-specific paint delineated the whole X chromosome of the red panda. The results indicate that at least 28 fissions (breaks), 49 fusions and 4 inversions were needed to "convert" the dog karyotype to that of the red panda, suggesting that extensive chromosome rearrangements differentiate the karyotypes of red panda and dog. Based on the established comparative chromosome homologies of dog and domestic cat, we could infer that there were 26 segments of conserved synteny between red panda and domestic cat. Comparative analysis of the distribution patterns of conserved segments defined by dog paints in red panda and domestic cat genomes revealed at least 2 cryptic inversions in two large chromosomal regions of conserved synteny between red panda and domestic cat. The karyotype of red panda shows high degree of homology with that of domestic cat. PMID- 11901995 TI - [Introduction of foreign DNA into the genome of rare minnow by sperm electroporation]. AB - Gene transfer was investigated in rare minnow via electroporated sperm. The sperm of the fish mixed with linear DNA (pCAhLFc) was electroporated. Then mature eggs were in vitro fertilized with these sperm cells. The DNA was extracted from the fries and analyzed by PCR. The percentage of fries with foreign genes varied between 25.5% and 66.7%. The observation of the electroporated sperm under microscope indicates that both the vigor of sperms and the successful ratio of the fertilization diminish, indicating that the electroporation with various parameters can hurt the sperm on different extent. The sperm cells electroporated with foreign genes, then was treated by DNA, and the DNA was extracted and was analyzed by PCR. It was found that foreign gene still exists, which approves that the electroporation can force the foreign gene enter the sperm of rare minnow. PMID- 11901996 TI - Comparative allozyme analysis of severel grasshopper species. AB - The allele frequency of four allozyme loci in four grasshopper species from two families of Catantopidae and Oedipodidae was examined using horizontal starch gel electrophoresis. The zymograms show that all four species have two loci in malate dehydrogenase (MDH). At MDH-1 one moderately migrated allele is shared and dominant in all four species. Locusta migratoria manilensi has two allele fixations in lactate dehydrogenase (LDH) and malic enzyme (ME); while Gastrimargus saussure has a fixed allele at MDH-1 and a unique fixation at MDH-2. The overall genetic variance was the highest in Oxya chinensis (Allele per locus = 3.0; He = 0.22), but the lowest in L. migratoria manilens (Allele per locus = 1.5; He = 0.013). Allozyme data suggests the four species are close in phylogenetic relationship, but differentiated in genetic variation levels. PMID- 11901998 TI - Genetic analysis of the panicle traits related to yield sink size of rice. AB - In the study, ten panicle traits associated with yield sink size were measured in a recombinant inbred population derived from Zhenshan 97 x Minghui 63. Generally, spikelets per panicle were more closely correlated with number of secondary branch per panicle, spikelets on secondary branch per secondary branch, and spikelet density. A total of 53 QTL were detected for ten traits in two years. Approximately 43.4% QTLs were detected in both two years, suggesting environmental effects on traits. Five chromosomal regions (G359-RG532 and C567 C86-RG236 on chromosome 1, R712-RM29 on chromosome 2, P-RG424 on chromosome 6, C148-RM258 on chromosome 10) were detected to have effects on multiple panicle traits. QTLs for traits, which were correlated, were generally localized in similar chromosomal regions, suggesting that pleiotropy and (or) linkage are the molecular basis of relationship between them. A large number of digenic interactions were detected, 18.2% of which were detected simultaneously in both two years. The proportion of common interactions was trait-depended, ranging 8.7% for spikelets on secondary branch per secondary branch to 32.6% for panicle length. Approximately 26.7% of common two-locus combinations had pleiotropic effects by simultaneously influencing two or more traits. Overall, the results indicate that each panicle trait is controlled by several QTLs, genotype x environment interaction, and a large number of epistatic interactions. PMID- 11901997 TI - [Identification of a resistance gene to bacterial blight (Xanthomonas oryzae pv. oryzae) in a somaclonal mutant HX-3 of indica rice]. AB - Using the mature embryo of a susceptible rice variety Minghui 63 as the explant, we have obtained a somaclonal mutant HX-3 through selection in vitro, which has showed resistance to bacterial blight. In 8 successive years, the resistance of R1 to R9 generations of HX-3 was identified by ZJ173, a typical bacterial blight strain in Yangtsu River valley, and the results showed that the resistance of HX 3 was stable and heritable. Genetic analysis also indicated that the resistance of HX-3 to bacterial blight was under a dominant gene controlling. Using 32 bacterial blight strains collected in China, Philippines and Japan, the resistance spectrum of HX-3 and other 13 testers with different major dominant resistance genes were tested. Results of 2 years (1999-2000) experiment showed that HX-3 had a broad resistance spectrum, which seemed to be different with those of the other dominant resistance genes identified. Allelic tests were also conducted by crossing HX-3 with IRBB4, IRBB7, CBB12 and IRBB21, and the F2 populations of each of the 4 crosses demonstrated resistant and susceptible plant segregation, indicating that the resistance gene in HX-3 different from Xa-4, Xa 7, Xa-12 and Xa-21. All these results proved that there was a new resistance gene in HX-3. We have designated the new gene as Xa-25(t). PMID- 11901999 TI - [Homoeologous grouping of R. kamoji chromosomes introduced in wheat using RFLP molecular marks]. AB - Twenty six DNA probes from seven homoeologous groups of triticeae were screened to reveal the RFLP between 45 wheat-R. kamoji derivatives and their parents R. kamoji, Chinese Spring, Yangmai 5. The result showed that the introduced R. kamoji chromosomes in 16 wheat-R. kamoji alien chromosome lines including additions, substitution or putative translocations were grouped into homoeologous group 1, 3, 5, 6 and 7. Alien chromosome pairs could be readily transmitted into the descendants. The added chromosomes in K139, K141, K214, K218, K219 and K224 disomic addition lines were grouped into homoeologous group 1, but the added chromosomes in K214 and K218 were different from K219 and K224 which originated from different genomes of R. kamoji. Ditelosomic addition line K147 might involve a R. kamoji chromosome long arm homoeologous to group 1 of wheat, and the added R. kamoji 1 L chromosomes in K139, K141 and K147 probably derived from different three genomes of R. kamoji. U chromosome of R. kamoji. showed homology to wheat homoeologous group 1. Homoeologous group 1 chromosome of R. kamoji, particularly its long arm is related to genes for scab resistance. Result also demonstrated a possible rearrangement occurred between homoeologous group 1 and group 6 of R. kamoji. Two R. kamoji chromosomes introduced in K203 were grouped to homoeologous group 1 and 6, respectively. In K166, the introgressed R. kamoji chromosome involved the short arm of group 5. Another alien chromosome line K177 was revealed as to be with introduced chromosome involving group 5L, 6S and 7SL of R. kamoji. Results also confirmed the homoeology between S, H and Y genomes of R. kamoji. PMID- 11902000 TI - [Cloning and characterization of D-113 gene promoter from cotton]. AB - To study the expression of late embryogenesis abundant gene in seeds, the 1,024 bp 5' flanking sequence of D-113 gene, a late embryogenesis abundant gene of Gossypium hirsutum cv. Coker 312, was cloned by PCR. The similarity compared with the sequence of Lea protein gene family published was 92.50%. There are three putative ABREs and one enhancer-like which riches A/T in the promoter. The promoter was fused to the beta-glucuronidase gene to form pLD II. Via a particle bombardment, pLD II was introduced into embryogenic calli of cotton and seeds of Brassica napus which were all treated with abscisic acid for 3d before bombardment, also into roots, stems and leafs of cotton. Transient expression was measured histochemically as spot number 24 h after bombardment. GUS sexpression was observed in the seeds of Brassica napus and the embryogenic calli of cotton, but not found in roots and leaves of cotton. Those results indicated that the expression of D-113 gene promoter was embryo specific. PMID- 11902001 TI - Growth and differentiation of transgenic callus regulated by phytohormones and antibiotics in transformation of loblolly pine. AB - Mature zygotic embryos of loblolly pine (Pinus taeda L.) were transformed by Agrobacterium tumefaciens strain LBA 4404 harbouring the plasmid pBI121 which carried the selectable marker gene, neomycin phosphotransferase II (npt II) controlled by the promoter of the nopaline synthase gene, and the uidA reporter gene, encoding beta-glucuronidase (GUS) driven by the cauliflower mosaic virus 35S promoter. Organogenic transgenic calli and transgenic regenerated plantlets were produced on selection medium containing 15 mg/L kanamycin, and confirmed by GUS histochemical staining, polymerase chain reaction (PCR), and southern blot analysis. Influences of phytohormone (BA/IBA) and antibiotics on growth and differentiation of organogenic transgenic calli were investigated. Of the phytohormone (BA/IBA) and antibiotics administered, 500 mg/L carbenicillin combined with 2 mg/L BA and 0.5 mg/L IBA (BA/IBA = 4) resulted in a 54.2% higher increase in the growth of transgenic calli as well as in the differentiation of transgenic calli, which was 45.7% more than that of control on the 6th week of culture. Claforan at 500 mg/L combined with 2 mg/L BA and 0.5 mg/L IBA resulted in a 40.8% increase in the growth of transgenic calli, and 38.7% increase in the frequency of transgenic calli forming adventitious shoots compared with the control. The growth and differentiation of transgenic calli of loblolly pine was reduced preferentially by higher BA/IBA (BA/IBA = 8), as well as high concentration of antibiotics (carbenicillin and claforan, 550 mg/L each). But it was observed that 450 mg/L and 500 mg/L carbenicillin and claforan caused an increase in growth and differentiation of transgenic calli. These results suggested that the establishment of an efficient Agrobacterium tumefaciens mediated transformation protocol for stable integration of foreign genes into loblolly pine was also dependent on the regulation of phytohormone and antibiotic on growth and differentiation of transgenic calli. This work could be useful for the future studies of genetic transformation of conifers. PMID- 11902003 TI - [A study of the effects on the symbiotic nitrogen fixation of Sinorhizobium fredii with the introduction of dctABD and nifA genes]. AB - A recombinant plasmid pHN307 containing C4-dicarboxylic acid transport genes (dctABD) from Sinorhizobium meliloti, nifA genes from Klebsiella pneumoniae and reporter genes luxAB from pDB30 was constructed by using pTR102 as the vector. The pHN307 was then introduced into the S. fredii HN01, YC4 and GR3 by tri parental mating, and the stability of pHN307 in the transconjugants under free living and symbiotic condition was also investigated. The results of plant pot experiment indicated that the introduction of pHN307 in the transconjugants could significantly increase the nodule fresh weight, shoot dry weight (biomass) and total nitrogen content of the symbiotic plants with soybean variety of Heilong 33. When the transconjugants were in symbiotic with soybean variety of Chuanzao No. 1, HN01 (pHN307) could significantly increase its root nodule number and fresh weight; GR3 (pHN307) could significantly increase its root nodule number, nodule fresh weight, shoot dry weight and total nitrogen content, but YC4(pHN307) demonstrated negative effect under the same condition. The results of this study suggested that the introduction of dctABD and nifA could improve the symbiotic nitrogen fixation efficiency and nodulation ability of the rhizobia tested, respectively, and its effect was relevant to the characteristics of recipient rhizobia and soybean variety. PMID- 11902002 TI - [Screening and detecting of proteins interaction with KyoT]. AB - To study the function of KyoT2 in vivo, two-hybrid yeast, purification of KyoT2 protein, preparation of antibody and GST-pulldown methods were used in the experiments. 42 clones were obtained after 5 x 10(6) clones were screened by four kinds of nutrition limitation and beta-galactosidase assay, 22 clones were obtained after restriction of positive clones. Finally, 13 genes were obtained by sequence assay. Two of these were RBP-Jk and PIAS1. After they and KyoT2 changed vectors, negative two-hybrid yeast was finished. The result was positive; Using KyoT2 protein and antibody GST-pulldown of KyoT2 and RBP-Jk, KyoT2 and PIAS1 were done, the result was also positive. Therefore, KyoT2 interacted with RBP-Jk and PIAS1. PMID- 11902004 TI - [The polymorphism of four Y-STR loci and its application in forensic medicine]. AB - We have obtained the alleles and haplotype distribution of 100 unrelated males from the Chinese Han and Zhuang populations at the four Y chromosome-specific loci A10, C4, A7.1, A7.2 by fluorescent primers and with 377 DNA sequencer. We observed 7, 6, 6, 6 alleles in loci A10, C4, A7.1, A7.2 respectively, the gene diversity (GD) values are 0.7776/0.629, 0.773/0.732, 0.5978/0.7272, 0.6664/0.6458 (Zhuang/Han). 114 haplotypes were found in 200 males, haplotype diversity (HD) values are 0.9786, 0.9772 (Zhuang/Han). We have confirmed the core repeats and its repeating number of these alleles by sequencing. A tetraplex PCR system consisting of the A10, C4, A7.1 and A7.2 loci was set up and it is of good specificity. We investigated the validation at these four Y-STRs loci in forensic application such as the male specificity, genetic stability and the sensitivity. In conclusion, the four Y-STRs loci are very suitable to forensic medicine analysis and paternity test. PMID- 11902006 TI - What price compromise? PMID- 11902005 TI - Clinical application of the tongue elevator. PMID- 11902007 TI - Molar control. Part 2. PMID- 11902008 TI - Effects of three electric toothbrushes on orthodontic bracket retention. PMID- 11902010 TI - A bite orthotic for the resting period between two phases of treatment. PMID- 11902009 TI - Precision distractor placement with a custom-made index. PMID- 11902011 TI - Lower incisor extraction treatment with the Invisalign system. PMID- 11902013 TI - A world for dental volunteers. PMID- 11902012 TI - The expanding requirement for informed consent in the practice of dentistry. PMID- 11902014 TI - PDA volunteers bring smiles to Healthlink. PMID- 11902015 TI - Obstacles to prostate cancer screening in African-American men. AB - The purpose of this correlational study was to measure structural obstacles to a free prostate cancer screening. The sample consisted of 549 men, 69% who were African-American. The men attended a prostate cancer educational program and were offered free prostate cancer screening at their physician of choice. Structural obstacles that were predictors in screening participation were "making an appointment" (p = 0.02), "planning for an appointment" (p = 0.05), and "reminders of prostate cancer screening" (p = 0.02). The demographic variables of race and marital status were also predictors of screening participation. Implications for health education are given. PMID- 11902016 TI - Self-esteem matters: racial & gender differences among rural southern adolescents. AB - Self-esteem does matter! It matters so much that Oprah dedicated an entire issue of "O" magazine to address the subject. "It's a woman's most treasured possession" (Winfrey, 2000a). Self-esteem has a profound influence on adolescent health promotion behaviors. This study contributes to understanding the role self esteem plays in the behavior of adolescents. Utilizing a secondary data analysis, race and gender self-esteem differences among adolescents were investigated. The sample of 1,237 students (46% African-American and 52% White) from rural southern areas consisted of 744 females and 493 males. Self-esteem was assessed using the Miller Self-esteem Questionnaire (SEQ). The Hendricks Perceptual Health Promoting Determinants Model (HPHD) provided the theoretical framework for the study. The results of the study revealed a statistically significant difference in various aspects of self-esteem according to race and gender. African-Americans and males had a higher self-esteem which is consistent with many prior studies. PMID- 11902017 TI - Examining medication knowledge and behavior of older African-American adult day care clients. AB - The purposes of this descriptive pilot study were to: 1) examine medication knowledge and behavior of older African-American adult day care clients; and, 2) describe how community health nurses working in adult day care centers can improve medication regimen compliance by utilizing the Rice Model of Dynamic Self Determination. Convenience sampling was used to recruit 40 older African Americans attending adult day care at two urban geriatric centers. Findings from this study suggested that the older African-Americans sampled needed assistance from adult children or other caregivers to follow their medication regimen as evidenced by not being able to open a pill bottle, or not having a system to take medications, having trouble swallowing their pills, and obtaining prescribed medications from the drug store. Further findings from this study also suggested that among these older African-American subjects that there was an enormous knowledge deficit regarding side effects of medications and usefulness of medications. In addition, the Rice Model was applied to a case study to illustrate how community heath nurses can improve medication regimen compliance for adult day care clients. PMID- 11902018 TI - Patterns of race and gender representation in health assessment textbooks. AB - With growing diversity in the nursing profession and in society, nursing educators face the challenge of selecting textbooks that promote cultural competence. A content analysis of photographs in nine health assessment textbooks was conducted in order to determine: 1) how accurately nurses and patients are presented in terms of race and gender; 2) how race and gender representations have changed over the last decade; and 3) how gender representations vary across different types of chapters within the texts. Although some recently published textbooks included a substantial proportion of photos of minority nurses, males and racial minorities were generally under-represented as nurses. Women and most ethnic minorities were consistently under-represented as patients. Female patients were also under-represented in cardiovascular and respiratory chapters and over-represented in chapters on reproduction. Nursing educators should screen visual images in textbooks for evidence of race and gender bias and inform publishers of the need for non-biased teaching materials. PMID- 11902019 TI - Use of focus groups for pain and quality of life assessment in adults with sickle cell disease. AB - This study assessed the effectiveness of using focus groups to obtain information about the characteristics of pain and quality of life in adults with sickle cell disease and their families. Five focus group sessions were held. Four groups consisted of adults diagnosed with sickle cell disease and one was composed of family members. Although focus groups were useful for addressing the purposes of the study, several volunteers in the patient group were unable to attend due to the occurrence of pain episodes. Adults with sickle cell disease identified recurring disabling pain and its consequences as interfering with their physical, emotional, and social quality of life. Several reported a pain aura that signaled the initiation of a painful episode. Useful self-care techniques and coping strategies were also identified. Common emotional responses included anger, hostility, depression, disenfranchisement, death anxiety and fatalism. The belief by those with sickle cell disease that health professionals viewed them as drug dependent often fueled angry and hostile responses. Religion was a major source for coping. Family members' quality of life was affected and they felt the negative impact of sickle cell disease on family relationships. Family members also reported feelings of helplessness, guilt, and parental self-blame. Health care providers could use information gained through this study to positively influence the care of adults with sickle cell disease. PMID- 11902020 TI - African-American women's perceptions of mammography screening. AB - While mammography has been shown to decrease breast cancer mortality, many African-American women are not receiving annual screenings. African-American women's reasons for not having mammograms are not well understood. This study therefore surveyed 164 African-American women concerning barriers to mammography screening. Outreach coordinators in two urban and one rural site in Texas asked African-American women to complete a checklist about the barriers to mammography screening. The 23-item Mammography Barriers Checklist, which was developed based upon one of the author's clinical experience and the research literature, included both internal and external barriers to screening. Women in all three geographic areas identified fear of finding cancer and mammography cost as the most important reasons for not having mammograms. These results suggest that outreach strategies that address fears related to mammography screening and help women find low-cost mammography resources may be more effective than those focusing strictly on providing information. PMID- 11902021 TI - Empowerment: a view of two low-income African-Americans communities. AB - Field theory and empowerment were used as guiding conceptual frameworks to address empowerment issues in two low-income inner city African-Americans communities. Field theory and empowerment provided a conceptualization of these communities in terms of the possible impact of the physical environment of these communities on their residents and the health care professionals who worked with these residents. The most likely response is learned helplessness and depression that are antithetical to empowerment. These frameworks also were helpful in generating strategies to foster empowerment among these community residents. These strategies include helping residents to redefine their behavior as ways of coping with a hostile environment that confronts them with poverty and racism and to reconnect with natural supports in the community for the purpose of enhancing community coalitions and alliances. PMID- 11902022 TI - The effects of socioeconomic status and increased body mass index on cardiovascular disease in African-American women. AB - Today within the United States, CVD is the leading cause of death in women with the highest mortality being seen in African-American women. A review of literature revealed that over the last decade, within the United States, there has been an overall reduction in the death rate due to CVD. However, the rate of decline has been less for women than for men and less for African-American women than for White women. Findings from some studies indicate that African-American women have increased risk factors as compared to other ethnic groups for CVD based upon conditions and behaviors affecting lifestyle. Fortunately, most of the CVD risk factors are modifiable and their occurrence can be widely prevented. Therefore, it is imperative that health care providers approach the issue of risk factors for CVD in African-American women as a heart disease epidemic. This approach is necessary if the United States is going to improve the health of all Americans, eliminate disparities, and improve the quality of life. PMID- 11902024 TI - Ending health disparities in African-American men: a clarion call to arms. PMID- 11902023 TI - An intervention to increase coping and reduce health care utilization for school age children and adolescents with sickle cell disease. AB - The purpose of this study was to examine whether coping with pain changed immediately and one year after a self-care intervention for school-age children and adolescents with sickle cell disease (SCD). Sixty-five children and 32 adolescents attended an educational program for living with SCD. They were then randomly assigned to relaxation, art therapy or attention-control groups. Coping was measured before, after the intervention, and 12 months later. Thirty-three children and 14 adolescents completed the one year follow-up. Although there was no significant increase in the overall number of coping strategies school-age children and adolescents used from baseline to 12 months, there was a significant increase in those strategies specifically targeted by the intervention. For adolescents, there was a significant increase in the total number of coping scores used before the intervention and one year later. When compared to well African-American adolescents, overall coping scores in this sample were significantly lower. Health care utilization related to clinic visits, emergency department visits and hospitalizations decreased significantly for all participants in the study. PMID- 11902025 TI - The development and demise of a cataract surgery database. AB - BACKGROUND: The American Academy of Ophthalmology (AAO; San Francisco) launched a national eye care outcomes initiative in 1996, in response to strong interest by third-party payers and managed care in performance measurement and outcomes. The AAO's outcomes initiative NEON (National Eyecare Outcomes Network) began with the design and launch of a prospective observational registry of patients undergoing cataract surgery. METHODS: Participants submitted a common set of data regarding patients' demographics, preoperative ophthalmologic history, physical exam, test results, functional status and symptoms, intraoperative procedures and events, and postoperative outcomes for all patients undergoing first or second eye cataract surgery. RESULTS: Between January 1, 1996, and March 30, 2001, a total of 249 ophthalmologists submitted data on 17,876 patients undergoing first or second eye surgery. All preoperative, intraoperative, and postoperative clinical data forms were submitted for 9,937 patients (55.7%). After surgery, 93% of patients achieved a best corrected visual acuity of 20/40, 89% improved their visual functioning, and 92% experienced fewer cataract-related symptoms. DISCUSSION: At the end of March 2001, NEON was discontinued because of a lack of participation and demand by members or third parties for this information. The momentum for accountability and performance measures never quite materialized into advantages for contracting for physicians or requirements by payers. In the future, more scientific evidence regarding the validity and meaning of outcome measures and differences in measurements, investment in health information technology infrastructure, use of technology to collect information at the point of care, and incentives favoring data collection and analysis will be needed to pave the way for renewed interest in outcomes. PMID- 11902026 TI - Private sector unlikely to follow Medicare lead in providing health plan disenrollment comparisons. AB - BACKGROUND: There have been substantial efforts to improve the measurement and reporting of comparative quality information. A three-stage effort to develop comparative voluntary disenrollment measures for private health insurance plans is described. The literature on disenrollment and how key groups might use disenrollment information is reviewed; the development of a comparative survey of disenrollment is described; reasons employers, purchasing coalitions, and plans were ultimately unwilling or unable to sponsor the survey are delineated; and implications of these findings are discussed. DATA AND METHODS: Methods used to develop the survey included review of existing literature on disenrollment, review of extant disenrollee surveys, cognitive testing, and expert review of the survey. Informal and formal interviews were conducted to assess the feasibility of different sponsors. RESULTS: A disenrollment survey instrument that covered areas of common interest to consumers, purchasers, and plans could be developed, but sponsors to test the collection and reporting of these data could not be recruited. This was due to four interrelated factors: technical challenges in developing appropriate samples, wide variation in resources and capabilities of purchasers and plans, the perception that the costs of the survey outweighed the benefits of comparative information on disenrollment to the different sponsors, and the absence of strong demand from purchasers, regulators, or consumers to motivate plans to collect or report comparative information on disenrollment. IMPLICATIONS: Several major barriers must be overcome before disenrollment information can become a component of comparative health care quality measures for the privately insured. PMID- 11902027 TI - Beyond rapid cycle: a one-day safety summit tool to prevent mislabeled laboratory specimens. PMID- 11902028 TI - Using comparison charts to assess performance measurement data. AB - BACKGROUND: In 1997 the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) announced the ORYX initiative, which integrates outcomes and other performance measurement data into the accreditation process. JCAHO uses control and comparison charts to identify performance trends and patterns that are provided to JCAHO surveyors in advance of a health care organization's (HCO's) survey. During the survey, the HCO is asked to explain its rationale for its selection of performance measures, how the ORYX data have been analyzed and used to improve performance, and the outcomes of these activities. CONSTRUCTING COMPARISON CHARTS: A comparison chart, a graphical summary of the comparison analysis, consists of actual (or observed) rates, expected rates, and expected ranges (upper and lower limits) for a given time frame. The expected range describes the degree of certainty that a given point is different from the average score (population). THE USE OF COMPARISON CHARTS: Comparison charts are primarily useful for telling an HCO whether one of its selected performance measures may be evidencing one of the three types of measurement outcomes: exemplary performance, average performance, or substandard performance (indicating an opportunity for improvement). The comparison charts compare an HCO's outcomes to those of its comparison group or to its risk-adjusted data. The charts provide guidance to an HCO about whether it should continue to monitor a process so as to maintain its current level of performance or whether it should try to improve its current performance. PMID- 11902029 TI - Improving children's health care: an interview with Charles Homer. Interview by Steven Berman. PMID- 11902032 TI - Needles threaten everyone. PMID- 11902031 TI - Men in nursing, unite! PMID- 11902030 TI - Eclampsia. PMID- 11902034 TI - Double-checking for errors. Lessons learned. PMID- 11902033 TI - Changing an ostomy appliance. PMID- 11902035 TI - Dietary supplement. Is kava safe? PMID- 11902036 TI - Extended-release drugs. Timing in everything. PMID- 11902037 TI - Inapsine. Weighing the risk of fatal arrhythmias. PMID- 11902038 TI - Arixtra injection. FDA approves synthetic anticlotting drug. PMID- 11902039 TI - Xeloda. Warning: drug interaction increase bleeding risks. PMID- 11902040 TI - Avelox. Respiratory antibiotic now in i.v. form. PMID- 11902041 TI - Microbe of the month. Streptococci viridans. PMID- 11902042 TI - Guarding your patient against ARDS. PMID- 11902043 TI - Steady the course of Parkinson's disease. PMID- 11902044 TI - Taking the measure of a father's grief. PMID- 11902045 TI - Writing easy-to-read teaching aids. PMID- 11902046 TI - Documenting for "PROs". PMID- 11902047 TI - Warming up to i.v. infusion. PMID- 11902048 TI - Reviewing common cardiovascular drugs. PMID- 11902049 TI - Latex allergy. Separating fact from fiction. PMID- 11902050 TI - Understanding hypokalemia. PMID- 11902051 TI - Patient falls. Walk with me. PMID- 11902052 TI - Brain death. Heart of the matter. PMID- 11902053 TI - Realistic expectations for safety devices. PMID- 11902054 TI - Understanding nociceptive pain. PMID- 11902055 TI - Myths & facts...about animal-assisted therapy. PMID- 11902056 TI - Understanding St. John's wort. PMID- 11902057 TI - Inching up. Annual sampling of clinical indicators for ESRD patients shows continuing improvement. PMID- 11902058 TI - Were patient voices heard in the technician certification debate in Georgia? PMID- 11902059 TI - Reflections on the optimal dialysis modality distribution: a North American perspective. PMID- 11902060 TI - The biological and economic value of oral organic iron in maintenance dialysis. AB - With the widespread use of recombinant erythropoietin (EPO) for patients with end stage renal disease (ESRD), management of iron deficiency is an ongoing issue for the renal team. Effective iron replacement and maintenance play a vital role in efficient use of EPO. For hemodialysis patients, intravenous (i.v.) iron has proven convenient and, as an ancillary drug outside of the composite rate, generates profits for dialysis facilities. Improvements in the vehicle with which i.v. iron is administered have led to a reduction in severe or fatal reactions common with iron dextran products. Oral iron has had a spotty track record as an effective therapy for dialysis patients. Compliance has been hindered by patient discomfort when taking oral iron. Patients on peritoneal dialysis and those with chronic kidney disease remain good candidates for oral iron because of convenience, and oral formulas could prove more effective even in the hemodialysis patient population if they were better tolerated and better absorbed, and if using them would not place an economic burden on the patient and/or an economic hardship on the facility. In a capitated/bundled payment environment, oral iron may become a blessing rather than a curse for facilities that need to find more economic ways of providing services. Heme-iron, now undergoing clinical studies, may be a reliable replacement for i.v. iron in that scenario. PMID- 11902061 TI - Heme iron polypeptide: a radically different oral iron. PMID- 11902062 TI - Billing for clinical social work services in dialysis facilities. PMID- 11902063 TI - NKF takes the next step in K/DOQI process with guidelines for CKD. PMID- 11902064 TI - Who will you call when disaster strikes in the dialysis facility? PMID- 11902065 TI - Protests continue over proposed vitamin D restrictions. PMID- 11902066 TI - Integration in nephrology. Life Options unveils new kidney disease paradigm. PMID- 11902067 TI - The value of active involvement. A CKD patient looks ahead. PMID- 11902068 TI - One CKD patient's journey. From kidney transplant back to dialysis. PMID- 11902069 TI - Exploring chronic kidney disease. An interview with Bryan Becker. PMID- 11902070 TI - Novel intraoperative cerebral blood flow monitoring by laser-Doppler scanner. AB - Laser-Doppler (LD) flowmetry was used to measure tissue perfusion non-invasively and continuously during neurosurgical operations using an LD scanner. Scanning was usually completed in 20 seconds. Measurements were processed in software to provide a color-coded image of the tissue perfusion. Moreover, the measurement data, expressed in LD-units, could be used for statistical data analysis. No physical contact was necessary between the scanning device and the exposed brain tissue. The imager provided two-dimensional microvascular flow maps non invasively and quantitatively during brain operations, and could show the CO2 reactivity in the vessels. LD scanning flowmetry is a promising intraoperative monitoring method for cerebral blood flow changes. PMID- 11902071 TI - Constructive interference in steady state imaging of moyamoya disease. AB - The diagnostic accuracy of three-dimensional constructive interference in steady state (CISS) magnetic resonance (MR) imaging was evaluated for the assessment of idiopathic moyamoya disease. Six consecutive patients underwent MR angiography, CISS imaging, and digital subtraction angiography. MR angiography and CISS imaging visualization of the steno-occlusive changes in the distal internal carotid arteries and the development of moyamoya vessels in the basal cistern were compared to the results obtained by digital subtraction angiography. MR angiography revealed the steno-occlusive changes correctly in nine and overestimated the changes in three of 12 hemispheres examined. CISS imaging showed the steno-occlusive changes defined as decreased caliber of the internal carotid artery correctly in two, underestimated the changes in nine, and overestimated the changes in one of the 12 hemispheres. MR angiography detected moyamoya vessels correctly in five and underestimated the vessels in seven of the 12 hemispheres. CISS imaging revealed the moyamoya vessels correctly in 10, underestimated the vessels in one, and overestimated the vessels in one of the 12 hemispheres. CISS imaging can supplement MR angiography in the non-invasive diagnosis of moyamoya disease, especially for the evaluation of moyamoya vessels. PMID- 11902072 TI - Transfemoral superior ophthalmic vein approach via the facial vein for the treatment of carotid-cavernous fistulas--two case reports. AB - Two elderly female patients with carotid-cavernous fistulas (CCFs) were treated by transvenous embolization through a transfemoral superior ophthalmic vein approach via the facial vein. Complete occlusion of CCFs was not achieved with this technique exclusively, but the technique was effective in these cases. This technique provides an alternative to other transvenous approaches for the treatment of CCFs. PMID- 11902073 TI - Ruptured aneurysm associated with partially duplicated posterior communicating artery--case report. AB - A 65-year-old woman presented with a ruptured saccular aneurysm associated with a rare variation of the posterior communicating artery (PcoA), partially duplicated PcoA. The PcoA with this variation forked just distal to the aneurysmal neck, and the two branches independently merged into the posterior cerebral artery. Initial clipping failed to isolate the aneurysm from one of the two branches, so the aneurysmal dome continued to pulsate and bleed. Temporary clipping of the proximal internal carotid artery revealed the fork of the two branches just distal to the aneurysmal neck. A curved Yasargil clip was used to clip the aneurysm and preserve the PcoA and branches. Careful observation of this PcoA variation is needed because the arterial structures may be hidden by the thickened arachnoid membrane. PMID- 11902074 TI - Primary hemangioma of the occipital bone in the region of the torcula--two case reports. AB - Two rare cases of subtorcular occipital bone hemangioma occurred in 26-year-old and 30-year-old female patients. Partial resection was performed in both cases because of the proximity to the torcula. No recurrence was seen at follow-up examination at 9 and 12 months. PMID- 11902075 TI - Presigmoid transpetrosal approach for the treatment of a large trochlear nerve schwannoma--case report. AB - A 61-year-old man presented with a rare, large trochlear nerve schwannoma manifesting as left-sided weakness and hypesthesia, bilateral bulbar pareses, and trochlear nerve paresis persisting for 3 months. T1-weighted magnetic resonance imaging with gadolinium revealed an intensely enhanced, well-circumscribed lesion with multicystic formation occupying the prepontine and interpeduncular cisterns and compressing the pons and midbrain with greater extension to the right. The mass was completely removed through the presigmoid transpetrosal approach with preservation of the posterior cerebral, superior cerebellar, and basilar arteries and their branches. Neuroradiological examination after 3 years demonstrated no recurrence. Enlargement of a tumor in the cisternal portion is inclined to involve and/or encase the adjacent major arteries and their branches. The presigmoid transpetrosal approach is one of the best surgical routes to remove a large trochlear nerve schwannoma safely and completely. PMID- 11902076 TI - Monostotic fronto-orbital fibrous dysplasia with convulsion--case report. AB - A 28-year-old man presented with monostotic fronto-orbital fibrous dysplasia associated with convulsions. Signs of meningeal irritation were observed. Computed tomography (CT) showed right frontal sinusitis, and destruction from the inner to outer table with expansion of the diploic space. T1- and T2-weighted magnetic resonance imaging showed an abnormal low-intensity mass, with heterogeneous gadolinium enhancement. Although the meningitis resolved, signs of infection continued for 2 months due to sinusitis. Treatment of the right frontal sinusitis was undertaken, accompanied by open biopsy. The histological diagnosis was fibrous dysplasia. Once the infection had completely resolved, orbitofrontal reconstruction was undertaken. Cranioplasty was carried out using cranial bone cement. Three-dimensional CT was valuable to show the likely postoperative result. PMID- 11902077 TI - Spinal cord compression due to extramedullary hematopoiesis associated with polycythemia vera--case report. AB - A 69-year-old woman with a 14-year history of polycythemia vera suffered progressive paraparesis due to epidural involvement of hematopoietic tissue. Magnetic resonance (MR) imaging demonstrated extensive epidural masses. Decompressive surgery and radiotherapy were performed and she made an almost complete clinical recovery. Serial MR imaging showed no regrowth of the other epidural masses. Extramedullary hematopoiesis occurs in patients with various hematologic disorders involving a chronic increase in the production of red blood cells, and is often associated with thalassemia, but is less common with polycythemia vera. The most frequent sites are the spleen, liver, and kidney. Extramedullary hematopoietic tissue occurring within the spinal canal and causing cord compression is very rare. Total surgical excision is not usually feasible because of the diffuse nature of extramedullary hematopoietic tissue and the possibility of recurrence, but acute neurological deterioration does require emergency surgery. Extramedullary hematopoiesis is radiosensitive and displays a rapid response to low dosages, so radiation therapy is recommended for residual tumors. Considering the possibility of central nervous system extramedullary hematopoiesis in patients with polycythemia vera, an early diagnosis is necessary for a favorable prognosis. PMID- 11902078 TI - Unusual arachnoid cyst of the quadrigeminal cistern in an adult presenting with apneic spells and normal pressure hydrocephalus--case report. AB - A 67-year-old woman was admitted to our clinic with symptoms of normal pressure hydrocephalus, lower cranial nerve pareses, and pyramidal and cerebellar signs associated with respiratory disturbances. Computed tomography (CT) and magnetic resonance imaging revealed a 4.7 x 5.4 cm quadrigeminal arachnoid cyst causing severe compression of the tectum and entire brain stem, aqueduct, and cerebellum, associated with moderate dilation of the third and lateral ventricles. Emergency surgery was undertaken due to sudden loss of consciousness and impaired breathing. The cyst was totally removed by midline suboccipital craniotomy in the prone position. Postoperatively, her symptoms improved except for the ataxia and impaired breathing. She was monitored cautiously for over 15 days. CT at discharge on the 18th postoperative day revealed decreased cyst size to 3.9 x 4.1 cm. Histological examination confirmed the diagnosis of the arachnoid cyst of the quadrigeminal cistern. The patient died of respiratory problems on the 5th day after discharge. Quadrigeminal arachnoid cysts may compress the brain stem and cause severe respiratory disturbances, which can be fatal due to apneic spells. Patients should be monitored continuously in the preoperative and postoperative period until the restoration of autonomous ventilation is achieved. PMID- 11902079 TI - Fast inversion recovery magnetic resonance imaging with the real reconstruction method: a diagnostic tool for cerebral gliomas. AB - The fast inversion recovery (IR) technique was evaluated for the localization of gliomas. Fast IR imaging with real reconstruction and T1-weighted spin echo (SE) imaging before and after contrast administration were performed in 20 patients with gliomas. The tumor-to-white matter contrast ratio (TWCR), tumor-to-gray matter contrast ratio (TGCR), tumor-to-white matter contrast-to-noise ratio (TWCNR), and tumor-to-gray matter contrast-to-noise ratio (TGCNR) were calculated and compared. Fast IR imaging visualized tumors with significantly higher TWCR, TGCR, TWCNR, and TGCNR values (p < 0.01) than those for T1-weighted SE imaging. In particular, fast IR imaging clearly revealed seven non-enhanced tumors that were poorly visualized on T1-weighted SE imaging. Fast IR imaging showed a similar TGCR and significantly higher TWCR (p < 0.01) compared to T1-weighted SE imaging with contrast medium in 13 enhanced tumors. However, fast IR imaging showed similar TWCNR and lower TGCNR compared to T1-weighted SE imaging with contrast medium. The fast IR technique can discriminate tumors from normal cerebral tissues with high contrast and without the use of contrast medium. This technique is extremely useful for the localization of non-enhanced tumors. PMID- 11902080 TI - Acute intestinal obstruction: diagnosis and management. AB - In acute intestinal obstruction, the clinician must distinguish between acute small bowel obstruction (ASBO) and acute colonic obstruction (ACO). In cases of ASBO, management depends on whether the patient has had previous abdominal surgery. Most cases of ACO require surgery, although mechanical causes must be distinguished from pseudo-obstruction for different management techniques. PMID- 11902081 TI - Diagnosis and immediate care of wrist and hand. PMID- 11902082 TI - eSTEP: a Web-based learning resource for basic surgical trainees. AB - A website has been set up by the Royal College of Surgeons of England for trainees registered on the Surgeons in Training Education Programme (STEP). eSTEP has been designed to provide an interactive component to the course and looks to have added a new dimension to basic surgical training. PMID- 11902083 TI - Mullerian duct cyst: a case history and literature review. PMID- 11902084 TI - Bony skull neoplasms masquerading as giant cell arteritis. PMID- 11902085 TI - Systemic lupus erythematosus presenting with brainstem lesions. PMID- 11902086 TI - Percutaneous balloon mitral valvuloplasty. PMID- 11902087 TI - Diagnosing hepatitis E in alcoholic patients. PMID- 11902088 TI - Managing schizophrenia. PMID- 11902089 TI - Saline or Hartmann's solution: is it still a controversy? PMID- 11902090 TI - Use of the bispectral index to monitor anaesthesia. PMID- 11902091 TI - Tonsillectomy and variant Creutzfeldt-Jakob disease. PMID- 11902092 TI - Pericardial constriction: diagnosis and management. AB - The diagnosis of pericardial constriction is challenging and elusive. It is a postinflammatory condition that occurs when a thickened, fibrotic, scarred and sometimes calcified pericardium firmly encases the cardiac chambers and restricts filling of the heart, causing venous overload and diminished cardiac output. This review includes the diagnosis and management of this condition. PMID- 11902093 TI - Treatment paradigms in heart failure. AB - Heart failure patients suffer very severe morbidity, high mortality and are common in medical practice. Their need for effective treatments is only partially answered by current options. Effective treatments often address cellular and pathophysiological mechanisms which are involved in the progression of heart failure. This article reviews those mechanisms and treatments. PMID- 11902094 TI - Lowering lipids after a stroke or transient ischaemic attack. AB - There is a lot of clinical uncertainty about how to aggressively pursue elevated cholesterol levels in patients following stroke or transient ischaemic attack. This article reviews the evidence linking cholesterol level with stroke and looks at whether treatment with lipid-lowering drugs can be justified. PMID- 11902095 TI - Profound hypotension: ethical considerations. AB - The use of profound induced hypotension to provide better operating conditions for surgery is long established. However, it is a controversial technique and it may be argued that it is inappropriate in modern anaesthetic practice. A currently used technique is reviewed against the benchmark of a lawsuit concerning profound hypotension. PMID- 11902096 TI - Restoring post-prandial insulin release in type 2 diabetes. AB - Tight blood glucose control is a primary aim of type 2 diabetes treatment. Combining metformin with the amino acid derivative, nateglinide, tackles both beta cell dysfunction and insulin resistance, and produces a greater decrease in haemoglobin A1c levels than treatment with either drug alone. PMID- 11902097 TI - Inflammatory responses after surgery. AB - The inflammatory response after major surgery is of great importance for patients, physicians and perioperative medicine in general. This article, although not intended to be comprehensive, provides an overview of present knowledge about inflammatory mechanisms, predictive parameters and therapeutic approaches. PMID- 11902098 TI - [Accuracy of magnetic resonance tomography in the diagnosis of meniscal tears]. PMID- 11902099 TI - [Rhinitis in old age]. AB - The chronic rhinitis is a frequent problem with age. In the nose, normal physiologic changes of aging include loss of nasal tip support, atrophy of mucus producing mucosal glands, and decreased olfaction. The fragmentation and weakening of the cartilage of the septum also causes airflow changes contributing to nasal stuffiness. These changes contribute to geriatric rhinitis, the symptoms of which are often attributed by the older patient to "allergies" or "sinus trouble". An understanding of these anatomic changes, linked with a thorough history and physical examination, allows the physician to properly manage geriatric rhinitis. Earnest causes like tumors or mycosis must be excluded. The treatment depends on the etiology of the rhinitis and also the symptoms of the patient. Therapy includes avoidance of all sorts of allergens (dust, fumes, sprays) and appropriate attention to minimizing home environmental and occupational exposures. Pharmacotherapy most often involves liquifying--not drying--nasal secretions with oral and topical preparations. Conservative surgical treatment is occasionally indicated. PMID- 11902100 TI - [Early-onset generalized polyarthritis (Stickler syndrome)]. AB - A 25-years old woman complained of pain especially in the knees and hips since she was 11 years old. Her pain worsened with weight bearing and activity. Aged 16 she underwent a transposition osteotomy of the right femoral neck. Clinical and radiographic examination demonstrated an early-onset polytopic osteoarthritis. The molecular DNA analysis confirmed the diagnosis of Stickler syndrome. The present case report demonstrates the importance of early recognition of the diagnosis and the management of osteoarthritis. PMID- 11902101 TI - [Vaginal granulocytic sarcoma: CT and MR imaging]. AB - A 30-year-old female patient with vaginal bleeding was referred to the gynecological unit of our hospital. Speculum examination showed a lobulated tumor, 5 cm in size, at the vaginal fornix. MRI demonstrated a tumor encompassing the ventral part of the vagina and the entire cervix. Computed tomography diagnosed pathologically enlarged mediastinal lymph nodes. Subsequent examinations revealed an acute myeloic leukemia, synchronous histopathological examination of the vaginal tumor led to the rare diagnosis of a granulocytic sarcoma. PMID- 11902102 TI - [Dunbar syndrome]. PMID- 11902103 TI - [Sarcoidosis of the submandibular gland]. PMID- 11902104 TI - Progress towards poliomyelitis eradication, Egypt, 2001. PMID- 11902105 TI - Influenza. PMID- 11902106 TI - Proceedings of the Conference of the European Society of Veterinary and Comparative Nutrition. Amsterdam, 24 April 2000. PMID- 11902107 TI - Estrogen doesn't prevent second strokes. PMID- 11902108 TI - Mechanisms of toxicity, carcinogenesis, cancer prevention, and cancer therapy. Abstracts of the 16th Aspen Cancer Conference. July 15-18, 2001. Aspen, Colorado. PMID- 11902109 TI - Protein misfolding and disease. Papers of a discussion meeting at the Royal Society on 23 and 24 February 2000. PMID- 11902110 TI - [118th Congress of the German Society of Surgery. 1-5 May 2001. Abstracts]. PMID- 11902111 TI - Calcium-induced cytochrome c release from CNS mitochondria is associated with the permeability transition and rupture of the outer membrane. AB - The mechanisms of Ca2+-induced release of Cytochrome c (Cyt c) from rat brain mitochondria were examined quantitatively using a capture ELISA. In 75 or 125 mm KCl-based media 1.4 micromol Ca2+/mg protein caused depolarization and mitochondrial swelling. However, this resulted in partial Cyt c release only in 75 mm KCl. The release was inhibited by Ru360, an inhibitor of the Ca2+ uniporter, and by cyclosporin A plus ADP, a combination of mitochondrial permeability transition inhibitors. Transmission electron microscopy (TEM) revealed that Ca2+-induced swelling caused rupture of the outer membrane only in 75 mm KCl. Koenig's polyanion, an inhibitor of mitochondrial porin (VDAC), enhanced swelling and amplified Cyt c release. Dextran T70 that is known to enhance mitochondrial contact site formation did not prevent Cyt c release. Exposure of cultured cortical neurons to 500 microM glutamate for 5 min caused Cyt c release into the cytosol 30 min after glutamate removal. MK-801 or CsA inhibited this release. Thus, the release of Cyt c from CNS mitochondria induced by Ca2+ in vitro as well as in situ involved the mPT and appeared to require the rupture of the outer membrane. PMID- 11902112 TI - Dopamine D4 receptors are heterogeneously distributed in the striosomes/matrix compartments of the striatum. AB - Two important aspects of striatal function, exploratory behaviour and motor co ordination, require the integrity of the dopamine D4 receptor subtype. These receptors are also implicated in the pathophysiology of certain neuropsychiatric disorders. However, the distribution of D4 receptors in the striatum has not yet been described and this situation impairs our understanding of the anatomical substrate in which D4 receptors function. We developed a D4 receptor-specific antibody that has permitted us to investigate the regional and cellular localization of the receptor in the neostriatum of the rat, mouse, cat and monkey. The subcellular distribution and the synaptic organization of this receptor were also determined in the rat striatum. We found moderate levels of D4 receptor expression in the caudoputamen and lower levels in the nucleus accumbens. These receptors were expressed in cell bodies and in the neuropil and were heterogeneously distributed among different striatal compartments, being more abundant in striosomes than in the matrix. At the subcellular level, the receptor immunoreactivity was mainly localized to dendritic shafts and spines. The prominent immunoreactivity observed in the striosomes indicates that integrative processes involved in D4-mediated limbic behaviours occurs through the striosomes rather than accumbens, whereas the motor behaviour is based in the striatal matrix. PMID- 11902113 TI - Essential role of caspase-11 in activation-induced cell death of rat astrocytes. AB - We have previously shown that rat astrocytes undergo apoptosis upon inflammatory activation. Nitric oxide (NO) produced by activated astrocytes was the major cytotoxic mediator in this type of autoregulatory apoptosis. However, an inhibitor of nitric oxide synthase did not completely block the apoptosis of activated astrocytes, suggesting the presence of other apoptotic pathways. Here, we present evidence that caspase-11 is an essential molecule in NO-independent apoptotic pathway of activated astrocytes. Inflammatory activation (lipopolysaccharide, interferon-gamma, and tumor necrosis factor-alpha treatment) of rat astrocyte cultures and C6 glioma cells led to the induction of caspase-11 followed by activation of caspases-11, -1, and -3. In contrast, NO donors induced activation of caspase-3 only. Inactivation of caspase-11 by the transfection of dominant negative mutant or treatment with the caspase inhibitors rendered the astrocytes partially resistant to the apoptosis following inflammatory activation, but not NO donor exposure. These results indicate that inflammatory stimuli not only induce the production of cytotoxic NO, but also initiate NO independent apoptotic pathway through the induction of caspase-11 expression. PMID- 11902115 TI - A new role for apolipoprotein E: modulating transport of polyunsaturated phospholipid molecular species in synaptic plasma membranes. AB - Phospholipids and their acyl group composition are important in providing the proper membrane environment for membrane protein structure and function. In particular, the highly unsaturated phospholipids in synaptic plasma membranes in the CNS are known to play an important role in modulating receptor function and neurotransmitter release processes. Apolipoprotein E (apoE) is a major apolipoprotein in the CNS, mediating the transport of cholesterol, phospholipids and their fatty acids, particularly in reparative mechanisms during neuronal injury. This study was performed to determine whether deficiency in the apoE gene contributes to an alteration of the phospholipids in synaptic plasma membranes. Phospholipid molecular species were identified and quantitated by HPLC/electrospray ionization-mass spectrometry. Analysis of the different phospholipid classes in membranes of apoE-deficient and C57BL/6 J mice indicated no obvious differences in the distribution of different phospholipid classes but substantial differences in composition of phospholipid molecular species. Of special interest was the prevalence of phospholipids (phosphatidylcholine, diacyl phosphatidylethanolamine, and phosphatidylserine) with 22:6n-3 in both the sn-1 and sn-2 positions of SPM and these phospholipid species were significantly higher in apoE-deficient mice as compared to control mice. Since polyunsaturated fatty acids in neurons are mainly supplied by astrocytes, these results revealed a new role for apoE in regulating polyunsaturated phospholipid molecular species in neuronal membranes. PMID- 11902114 TI - Phosphatidylinositol 3-kinase is a central mediator of NMDA receptor signalling to MAP kinase (Erk1/2), Akt/PKB and CREB in striatal neurones. AB - Ca2+ influx through NMDA receptors can initiate molecular changes in neurones which may underlie synaptic plasticity, neuronal development, survival and excitotoxicity. Signalling through the MAP kinase (Erk1/2) cascade may be central to these processes. We previously demonstrated that Ca2+-permeable AMPA receptors activate Erkl/2 through a phosphatidylinositol 3-kinase (PI 3-kinase)-dependent mechanism. We now report that NMDA receptor activation of Erk1/2 was also blocked by inhibitors of PI 3-kinase (LY 294002, wortmannin). In addition, pre-treatment of neurones with pertussis toxin inhibited NMDA-induced Erk1/2 activation, indicating a role for heterotrimeric Gi/o proteins. PI 3-kinase directs activation of the serine-threonine kinase Akt (PKB). Treatment of striatal neurones with glutamate induced a rapid Ca2+-dependent and PI 3-kinase-dependent phosphorylation of Akt (Ser473), which was not blocked by the Mek inhibitors PD98059 or U0126. Targets for Erk1/2 and Akt pathways include transcription factors. Glutamate-induced phosphorylation of cAMP response element binding protein (CREB; Ser133) was partially blocked with either PD98059, U0126, LY294002 or wortmannin but was very strongly inhibited on co-application of LY294002 and PD98059. We propose that NMDA receptor stimulation can activate Erk1/2 and Akt signalling pathways in a PI 3-kinase dependent manner which may target CREB in the nucleus. PMID- 11902116 TI - Neuroprotection by adenosine A2A receptor blockade in experimental models of Parkinson's disease. AB - Adenosine A2A receptors are abundant in the caudate-putamen and involved in the motor control in several species. In MPTP-treated monkeys, A2A receptor-blockade with an antagonist alleviates parkinsonian symptoms without provoking dyskinesia, suggesting this receptor may offer a new target for the antisymptomatic therapy of Parkinson's disease. In the present study, a significant neuroprotective effect of A2A receptor antagonists is shown in experimental models of Parkinson's disease. Oral administration of A2A receptor antagonists protected against the loss of nigral dopaminergic neuronal cells induced by 6-hydroxydopamine in rats. A2A antagonists also prevented the functional loss of dopaminergic nerve terminals in the striatum and the ensuing gliosis caused by MPTP in mice. The neuroprotective property of A2A receptor antagonists may be exerted by altering the packaging of these neurotoxins into vesicles, thus reducing their effective intracellular concentration. We therefore conclude that the adenosine A2A receptor may provide a novel target for the long-term medication of Parkinson's disease, because blockade of this receptor exerts both acutely antisymptomatic and chronically neuroprotective activities. PMID- 11902117 TI - Endogenous morphine modulates acute thermonociception in mice. AB - The endogenous synthesis of morphine has been clearly demonstrated throughout the phylogenesis of the nervous system of mammals and lower animals. Endogenous morphine, serving as either a neurotransmitter or neurohormone, has been demonstrated in the nervous system of both vertebrates and invertebrates. As one of the effects of exogenous morphine is the modulation of pain perception, we investigated the effects that the depletion of endogenous morphine had on nociceptive transmission. The immunoneutralization of endogenous morphine from brain extracellular spaces was obtained through the intracerebroventricular administration of affinity purified anti-morphine IgG to mice, which then underwent the hot plate test. Endogenous morphine immunoneutralization decreased thermal response latency and attenuated the anti-nociceptive effect of the mu selective agonist DAMGO in hot plate test suggesting that endogenous morphine is involved in pain modulation. PMID- 11902118 TI - Effect of endogenous serotonin on the binding of the 5-hT1A PET ligand 18F-MPPF in the rat hippocampus: kinetic beta measurements combined with microdialysis. AB - By using a combination of an original beta+-sensitive intracerebral probe and microdialysis, the effect of increased endogenous serotonin on specific binding of 18F-MPPF [4-(2'-methoxyphenyl)-1-[2'-[N-(2"-pyridinyl)-p fluorobenzamido]ethyl]piperazine] to the serotonin-1A (5-HT1A) receptors was investigated in the hippocampus of the anaesthetized rat. Our beta-sensitive probe prototype was sensitive enough to obtain specific 18F-MPPF time-activity curves in the rodent (hippocampus/cerebellum ratio approximately 2). The serotonin neuronal release was pharmacologically enhanced using fenfluramine at three different doses (1, 2 and 10 mg/kg intravenous) multiplying by 2-15 the extracellular serotonin in the hippocampus. These extracellular variations of extracellular serotonin resulted in dose-ranging decreases in 18F-MPPF-specific binding in the same rat. Our results showed for the first time that 18F-MPPF binding could be modulated by modifications of extracellular serotonin in the rat hippocampus. These results were confirmed by the enhancement of extracellular radioactivity collected in dialysates after the displacement of 18F-MPPF by fenfluramine. After modelization, 18F-MPPF binding could constitute an interesting radiotracer for positron emission tomography in evaluating the serotonin endogenous levels in limbic areas of the human brain. PMID- 11902119 TI - Functional expression of corticotropin-releasing hormone (CRH) receptor 1 in cultured rat microglia. AB - Corticotropin-releasing hormone (CRH), known as a key regulator of the hypothalamic-pituitary-adrenal axis response to stress, elicits its biological effects by binding to two membrane receptors (CRH-R1 and CRH-R2). The present studies examined the presence of functional expression of CRH receptors in cultured microglia of rat. CRH-R1 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR), western blotting and receptor chemical cross-linking assay in cultured microglia. CRH-R2 mRNA was undetectable by RT-PCR. The radioligand binding analysis using [125I]Tyr-rat/human CRH revealed a high affinity binding site (Kd of 1.2 nm and Bmax of 84 fmol/mg of protein). Competition studies using CRH and related peptides indicated kinetic and pharmacological characteristics consistent with the CRH-R1 receptor subtype. Receptor chemical cross-linking assay demonstrated a single band of CRH receptor with a molecular weight of -77 kDa, which was inhibited in the presence of excess unlabeled rat/human CRH in a dose-dependent manner and inhibited by a CRH receptor antagonist astressin. Functional coupled cAMP production in cultured microglia was stimulated by exogenous addition of CRH and related peptides in a dose-dependent manner and blocked by astressin. Our findings suggest the functional expression of CRH-R1 receptor in rat microglia, indicating an important mechanism of interaction between immune and neuroendocrine systems in brain physiological and pathological conditions. PMID- 11902120 TI - Overexpression of c-Fos is sufficient to stimulate tyrosine hydroxylase (TH) gene transcription in rat pheochromocytoma PC18 cells. AB - The AP1 site within the tyrosine hydroxylase gene proximal promoter is essential for the response of the gene to numerous stimuli. Stimulation of this gene is often associated with induction of the AP1 transcription factor, c-Fos. However, many stimuli activate or induce multiple transcription factors that interact with this AP1 site or other sites within the gene's proximal promoter. Hence, it remains unclear whether c-Fos induction by itself is sufficient to stimulate the tyrosine hydroxylase gene. In this study we produce rat pheochromocytoma PC18 cells that overexpress c-Fos under control of the tet-inducible system. We demonstrate that induction of c-Fos leads to dramatic stimulation of tyrosine hydroxylase gene transcription rate measured using nuclear run-on assays. This stimulation is closely associated quantitatively with the induction of c-Fos and does not apparently require phosphorylation of c-Fos. The response is partially dependent on the AP1 site within the tyrosine hydroxylase proximal promoter. However, the response of the proximal promoter to c-Fos induction is relatively small compared with that of the endogenous gene. Consequently, our results suggest that c-Fos exerts its influence on the tyrosine hydroxylase gene via multiple mechanisms that are dependent and independent of the proximal promoter AP1 site. PMID- 11902121 TI - Evaluation of the protective effect of oestradiol against toxicity induced by 6 hydroxydopamine and 1-methyl-4-phenylpyridinium ion (Mpp+) towards dopaminergic mesencephalic neurones in primary culture. AB - Recent findings suggest that gonadal steroid hormones are neuroprotective and may provide clinical benefits in delaying the development of Parkinson's disease. In this report we investigated the ability of oestradiol to protect mesencephalic dopaminergic neurones cultured in serum-free or serum-supplemented medium from toxicity induced by 6-hydroxydopamine or 1-methyl-4-phenylpyridinium ion (MPP+). The efficiency of both toxins and oestradiol was evaluated by tyrosine hydroxylase (TH) immunocytochemistry, [3H]dopamine ([3H]DA) uptake, length of dopaminergic processes and lactate dehydrogenase (LDH) release measurement. In cultures grown in serum-supplemented medium, a 2-h pre-treatment with high concentrations (10-100 microM) of 17beta-oestradiol or 17alpha-oestradiol, the stereoisomer with weak oestrogenic activity, protected both dopaminergic and non dopaminergic neurones from toxicity induced by 6-hydroxydopamine (6-OHDA; 40 or 100 microM) and by the high MPP+ concentrations (50 microM) necessary to obtain significant neuronal death under those culture conditions. At these concentrations, MPP+ was no longer selective for dopaminergic neurones but affected all cells present in the culture. In contrast, the hormonal treatments did not protect against selective degeneration of dopaminergic neurones induced by lower MPP+ concentrations (below 10 microM), related to inhibition of complex I of respiratory chain. In cultures grown in serum-free medium, oestradiol concentrations higher than 1 microM induced neuronal degeneration and no protection against 6-OHDA or MPP+ toxicity was observed at lower concentrations of the steroid. The neuroprotective effects of 17alpha- or 17beta-oestradiol evidenced in this model might be due to the antioxidant properties of these compounds. However, other non-genomic effects of the steroids cannot be excluded. PMID- 11902122 TI - Differential cell surface expression of GABAA receptor alpha1, alpha6, beta2 and beta3 subunits in cultured mouse cerebellar granule cells influence of cAMP activated signalling. AB - In this study we have used mature, primary cultured mouse cerebellar granule cells (CGCs) to initiate our studies on the mechanisms governing neuronal trafficking of GABAA receptors (GABARs). Initially the steady-state distribution of GABAR alpha1, alpha6, beta2 and beta3 subunits between the cell surface and cell interior was quantified. Cell surface proteins were modified with a membrane impermeable cross-linking agent, bis(sulfosuccinimidyl)suberate (BS3) or the proteolytic enzyme, chymotrypsin. The proportion of unmodified (intracellular) and modified (cell surface) subunits was quantified by immunoblotting. We found that 51% of alpha6, 74% of alpha1, and 83% of beta2/3 were expressed at the cell surface, thus identifying a sizeable intracellular pool of alpha6 in contrast to the low levels of intracellular alpha1 and beta2/3. Chronic activation of protein kinase A (PKA) in CGCs in vitro, post-transcriptionally up-regulated expression of alpha1, beta2 and beta3 but not beta6. This was paralleled by an increase in the BZ-S subtype of [3H]Ro154513 binding sites. GABAR alpha1 was increased at the cell surface and in the cell interior, beta2 was increased almost exclusively at the cell surface whilst beta3 was increased almost exclusively in the cell interior. The intracellular pool of alpha6 was not affected. Thus, GABAR subunits are subject to differentially regulated trafficking, affording yet greater scope for GABAR diversity and plasticity. PMID- 11902123 TI - Locally synthesized angiotensin modulates pineal melatonin generation. AB - We aimed to study the mechanisms and the significance of the influence exerted by the renin-angiotensin system (RAS) on the pineal melatonin production. Pineal melatonin and other indoles were determined by HPLC with electrochemical detection after angiotensin AT1-receptor blockade with Losartan in vivo or in cultured glands. N-acetyltransferase (NAT) activity was radiometricaly measured. To test the in vivo relevance of the local RAS, pineal melatonin and its indole precursors were determined in transgenic rats with inhibited production of angiotensinogen exclusively in astrocytes, TGR(ASrAOGEN). Tryptophan hydroxylase (TPH) and NAT mRNA levels were determined by real-time RT-PCR. Pineal melatonin content was significantly decreased by AT1-receptor blockade in vivo, in cultured glands and in TGR(ASrAOGEN) (35%, 32.4% and 17.5% from control, respectively). Losartan produced a significant decrease of pineal 5-hydroxytryptophan, serotonin, 5-hydroxyindole acetic acid and N-acetylserotonin in pineal cultures. Also, the pineal content of the precursor indoles in TGR(ASrAOGEN) rats was significantly lowered. The reduction of 5-hydroxytryptophan levels by 33-75% in both in vivo and in vitro studies suggests a decreased activity of TPH. Moreover, the TPH mRNA levels in TGR(ASrAOGEN) rats were significantly lower than control rats. On the other hand, NAT activity was unaffected by Losartan in pineal culture and its expression was not significantly different from control in TGR(ASrAOGEN) rats. Our results demonstrate that a local pineal RAS exerts a tonic modulation of indole synthesis by influencing the activity of TPH via AT1 receptors. PMID- 11902124 TI - Functional characterization of a novel serotonin receptor (5-HTap2) expressed in the CNS of Aplysia californica. AB - Serotonin has been shown to be a neuromodulator in the Aplysia californica CNS. The diversity of serotonin actions is due to the existence of several different receptor subtypes. In this study we report the cloning of a full-length cDNA, coding for a novel serotonin receptor (5-HTap2). The receptor protein bears the characteristics of G protein-coupled receptors. It shares 68% and 34% of its amino acid sequence identity with the 5-HTlym receptor from Lymnaea stagnalis and the mammalian 5-HT1A receptor, respectively. When transfected in HEK 293 cells, 5 HTap2 was negatively coupled to adenylate cyclase. Ligand binding analysis indicated that the order of potencies of various drugs for the inhibition of [3H]LSD binding was: methiothepin > metergoline > 5-CT > PAPP > 5-HT > ketanserin > NAN-190 > 8-OH-DPAT > clozapine. RT-PCR amplification of RNA isolated from different tissues indicated that this receptor is expressed in the CNS and in bag cells. The expression of 5-HTap2 restricted to the CNS suggests an important role for this receptor in the modulation of neuronal functions in Aplysia. Moreover, the high expression of 5-HTap2 in the bag cells, associated with its pharmacological profile, suggests that this receptor may be implicated in modulating the afterdischarge during the egg-laying behavior. PMID- 11902125 TI - A link between maze learning and hippocampal expression of neuroleukin and its receptor gp78. AB - Neuroleukin (NLK) is a multifunctional protein involved in neuronal growth and survival, cell motility and differentiation, and glucose metabolism. We report herein that hippocampal expression of NLK and its receptor gp78 is associated with maze learning in rats. First, mRNA levels of NLK and gp78 were significantly increased in hippocampi of male Fischer-344 rats following training in the Stone T-maze and the Morris water maze. Second, a parallel increase was found in hippocampal NLK and gp78 proteins after maze learning. Third, NLK and gp78 mRNA and protein expression in hippocampus was reduced in a group of aged rats that showed more errors during the acquisition of the Stone maze task as compared with young rats. Finally, application of recombinant NLK to hippocampal neurons significantly enhanced glutamate-induced ion currents, functional molecular changes that have been correlated with learning in vivo. Taken together, our results identify a novel association of hippocampal expression of NLK and its receptor gp78 with rat maze learning. Interaction of NLK with gp78 and subsequent signaling may strengthen synaptic mechanisms underlying learning and memory formation. PMID- 11902126 TI - Reduced nitric oxide metabolites in CSF of patients with tetrahydrobiopterin deficiency. AB - We investigated CSF concentrations of nitrite and nitrate as indicators of nitric oxide (NO) production in patients with tetrahydrobiopterin (BH4) deficiencies. Patients with 6-pyruvoyl-tetrahydropterin synthase, sepiapterin reductase and dihydropteridine reductase deficiencies exhibited decreased CSF nitrite + nitrate levels compared with healthy control subjects. Reduced levels of nitrite + nitrate were not influenced by oral administration of 2.5-5.0 mg/kg tetrahydrobiopterin. Our data indicate impaired NO synthase function in patients with BH4 deficiency and suggest possible involvement in the neuronal cell dysfunction. PMID- 11902128 TI - Bibliography. Current world literature. Epidemiology and health services research. PMID- 11902129 TI - Bibliography. Current world literature. Nonarticular rheumatism, sports-related injuries, and related conditions. PMID- 11902127 TI - Re: Oxygen in wound healing: more than a nutrient. PMID- 11902130 TI - The ACCC: what's it all about?. PMID- 11902131 TI - [A vesicular disease in a young woman]. PMID- 11902132 TI - [A feverish patient in good condition]. PMID- 11902133 TI - [Clinical relevance of polyclonal IgA gammopathy]. PMID- 11902134 TI - [Current status of diagnosis and therapy of Whipple's disease]. PMID- 11902135 TI - 2002 Trustees of the Year. Missouri lawyer brings a hospital to hometown. PMID- 11902136 TI - Transplantation-related toxicity and acute intestinal graft-versus-host disease after conditioning regimens intensified with Rhenium 188-labeled anti-CD66 monoclonal antibodies. PMID- 11902137 TI - Discrepancy between antithrombin activity methods revealed in Antithrombin Stockholm: do factor Xa-based methods overestimate antithrombin activity in some patients? PMID- 11902138 TI - RHCE represents the ancestral RH position, while RHD is the duplicated gene. PMID- 11902139 TI - Deletions of the derivative chromosome 9 do not account for the poor prognosis associated with Philadelphia-positive acute lymphoblastic leukemia. PMID- 11902140 TI - Does the P(2X1del) variant lacking 17 amino acids in its extracellular domain represent a relevant functional ion channel in platelets? PMID- 11902141 TI - Role of surface IgM and IgD on survival of the cells from B-cell chronic lymphocytic leukemia. PMID- 11902142 TI - Immunohistochemical localization of phosphorylated AKT in multiple myeloma. PMID- 11902143 TI - Anticoagulants for prophylaxis and treatment of thromboembolic events. PMID- 11902144 TI - Novel electrophysiologic parameter of dispersion of atrial repolarization: comparison of different atrial pacing methods. AB - INTRODUCTION: Heterogeneity of ventricular repolarization plays a major role in reentrant tachyarrhythmias in cardiac tissue. However, the role of atrial repolarization added activation time (AT) to refractoriness in atrial vulnerability has not been investigated in detail. METHODS AND RESULTS: The study population consisted of 34 patients: 18 with atrial fibrillation (AF) and 16 without AF (control group). The effective refractory periods (ERPs) in the right atrial appendage, low lateral right atrium, high right septum, and distal coronary sinus, and ATs from P wave onset to each electrogram during sinus rhythm and right atrial appendage, low lateral right atrial, high right septal, distal coronary sinus, and biatrial pacing were measured. Atrial recovery time, defined as the sum of AT and ERP, and its dispersions during sinus rhythm, right atrial appendage, low lateral right atrial, high right septal, distal coronary sinus, and biatrial pacing were calculated. Both ERP dispersion and atrial recovery time dispersion during sinus rhythm were significantly greater in the AF group than in the control group. Atrial recovery time dispersion during distal coronary sinus, high right septal, or biatrial pacing was significantly smaller than that during right atrial appendage or low lateral right atrial pacing in each group. In particular, atrial recovery time dispersion during distal coronary sinus pacing was the smallest of the five pacing methods in the AF group. P wave duration during biatrial or high right septal pacing was significantly shorter than during right atrial appendage, low lateral right atrial, or distal coronary sinus pacing in each group. CONCLUSION: Atrial recovery time dispersion is suitable as an electrophysiologic parameter of atrial vulnerability. Distal coronary sinus pacing may prevent AF by increasing homogeneity of atrial repolarization, whereas biatrial and high right septal pacing contribute not only homogeneity of atrial repolarization but also improvement of atrial depolarization. PMID- 11902146 TI - Determination of thickness, refractive index, and thickness irregularity for semiconductor thin films from transmission spectra. AB - A simplified theoretical model has been proposed to predict optical parameters such as thickness, thickness irregularity, refractive index, and extinction coefficient from transmission spectra. The proposed formula has been solved for thickness and thickness irregularity in the transparent region, and then the refractive index is calculated for the entire spectral region by use of the interference fringes order. The extinction coefficient is then calculated with the exact formula in the transparent region, and an appropriate model for the refractive index is used to solve for the extinction coefficient in the absorption region (where the interference fringes disappear). The proposed model is tested with the theoretical predicted data as well as experimental data. The calculation shows that the approximations used for solving a multiparameter nonlinear equation result in no significant errors. PMID- 11902145 TI - Clinical evaluation of a policy of early repeated internal cardioversion for recurrence of atrial fibrillation. AB - INTRODUCTION: The clinical value of cardioversion (CV) of persistent atrial fibrillation (AF) is limited by the high rate of early AF recurrence, which may be related to the persistence of atrial electrical remodeling. We examined the hypothesis that the likelihood of maintaining sinus rhythm after CV of persistent AF is significantly enhanced by a policy of early repeated CV. METHODS AND RESULTS: Fifty-nine patients with persistent AF underwent internal CV (CV 1). Those patients cardioverted were monitored with daily transtelephonic ECG. In the event of AF recurrence, these patients were admitted rapidly for repeat CV (CV 2) and, if further recurrence occurred, a third CV (CV 3) was performed. Daily ECG monitoring was continued until 1 month of sinus rhythm was maintained or a total of three CVs were performed. Of the 59 patients undergoing CV 1, 43 were discharged in sinus rhythm and 29 subsequently had AF recurrence during monitoring. Twenty-three of these underwent CV 2 and 11 of these underwent CV 3. Of those having repeated CVs, only 4 patients maintained sinus rhythm for 1 month (3 after CV 2 and 1 after CV 3). The remaining patients had repeated AF recurrence during the monitoring period. Mean time from AF recurrence to CV 2 was 20+/-13 hours and from AF recurrence to CV 3 was 13+/-7.2 hours. Atrial effective refractory periods increased from 189+/-16 msec at CV 1 to 215+/-18 msec at CV 3 (P < 0.05), indicating reversal of atrial electrical remodeling during this period. CONCLUSION: A policy of early repeated CVs for AF recurrence has very limited clinical value despite evidence of reversal of atrial electrical remodeling. The time between AF recurrence and repeat CV may need to be reduced further if such a policy is to succeed. PMID- 11902147 TI - Symmetrically coated pellicle beam splitters for dual quarter-wave retardation in reflection and transmission. AB - A trilayer pellicle that consists of a high-index center layer that is symmetrically coated on both sides by a low-index film can be designed to produce differential reflection and transmission phase shifts of +/- 90 degrees at oblique incidence and equal throughput for the p and the s polarizations. Such a device splits a beam of incident linearly polarized light into two orthogonal circularly polarized components that travel in well-separated angular directions. Examples of infrared dual quarter-wave retarders that use a symmetrically coated Ge pellicle at 77 degrees angle of incidence are presented. A 50-50% splitter requires a symmetric pellicle with at least five layers. Error analysis shows that the thicknesses of the high-index layers must be tightly controlled. These circular polarization beam splitters are intended for operation with a well collimated light source and can be used as the basis of a novel circular polarization Michelson interferometer. PMID- 11902148 TI - Active aberration correction for the writing of three-dimensional optical memory devices. AB - We describe an active optical system that both measures and corrects the aberrations introduced when writing three-dimensional bit-oriented optical memory by a two-photon absorption process. The system uses a ferroelectric liquid crystal spatial light modulator (FLCSLM) configured as an arbitrary wave-front generator that is reconfigurable at speeds as great as 2.5 kHz. A method of aberration measurement by the FLCSLM wave-front generator is described. The same device is also used to correct the induced aberrations by preshaping the wave fronts with the conjugate phase aberration as well as to scan the focal spot in three dimensions. Experimental results show the correction of both on- and off axis aberrations, allowing the writing of data at depths as great as 1 mm inside a LiNbO3 crystal. PMID- 11902149 TI - Prescriptions: why they're so complicated. PMID- 11902150 TI - [Fur color gene profiles and length of hair of several cat populations in Cuba, Costa Rica, Colombia, Paraguay, Chile and Argentina, and possible genetic origins of these cat populations]. AB - Until the present moment, only a scarce number of Latin American domestic cat populations have been studied from a population genetic standpoint. For this reason, the cat populations of La Havana (Cuba), San Jose (Costa Rica), Bogota and Ibague (Colombia), Asuncion (Paraguay), Santiago (Chile) and Buenos Aires (Argentina) were sampled for several coat genes. The results obtained were as follows: (1) there was a strong genetic resemblance between several Hispanic American cat populations (especially, those of Buenos Aires, San Jose and the two Colombian populations studied) and those from South Western United States (California, Texas and Colorado), which adds support to the suspicion that these populations probably have a common origin; (2) The cat population of Santiago (Chile), contrarily to the other Hispanic American populations studied, showed a strong genetic resemblance with some Anglo North American populations; and (3) The l (long hair) and d (dilution) alleles showed systematic higher frequencies in the Hispanic American populations than those observed in Spain. Although the Hispanic American populations were not identical to the current Spanish populations (with the exception of Asuncion), this historic genetic experiment was very different to that found for the British populations and their overseas colonies. PMID- 11902151 TI - Enhancing tumor implantation and growth rate of Ramos B-cell lymphoma in nude mice. AB - The ability of a human B-cell lymphoma cell line to grow subcutaneously as tumors in nude mice was investigated. The effect of pretreating mice with cyclophosphamide or whole-body irradiation (WBI) was compared with no pretreatment of the mice. Both methods of pretreatment resulted in a higher tumor implantation rate, compared with that for non-pretreated controls. In mice that underwent WBI-pretreatment, a tumor implantation rate of 100% was observed, whereas mice pretreated with cyclophosphamide had a tumor implantation rate of 80%. In non-pretreated control mice, an implantation rate of only 50% was observed. Three weeks after injection, tumor size was significantly larger in mice of the pretreated groups, compared with that in mice of the group that did not receive pretreatment. Furthermore, particularly in the group pretreated with WBI, the tumors grew more synchronously, compared with tumors in the control group. Results of this study indicate that pretreatment with cyclophosphamide or WBI improves the tumor implantation rate of Ramos cells in nude mice, providing a workable animal model for studying human B-cell lymphoma. PMID- 11902152 TI - Priming in implicit memory tasks: prior study causes enhanced discriminability, not only bias. AB - R. Ratcliff and G. McKoon (1995, 1996, 1997; R. Ratcliff, D. Allbritton, & G. McKoon, 1997) have argued that repetition priming effects are solely due to bias. They showed that prior study of the target resulted in a benefit in a later implicit memory task. However, prior study of a stimulus similar to the target resulted in a cost. The present study, using a 2-alternative forced-choice procedure, investigated the effect of prior study in an unbiased condition: Both alternatives were studied prior to their presentation in an implicit memory task. Contrary to a pure bias interpretation of priming, consistent evidence was obtained in 3 implicit memory tasks (word fragment completion, auditory word identification, and picture identification) that performance was better when both alternatives were studied than when neither alternative was studied. These results show that prior study results in enhanced discriminability, not only bias. PMID- 11902153 TI - False memory and the discrepancy-attribution hypothesis: the prototype familiarity illusion. AB - According to the discrepancy-attribution hypothesis (B. W. A. Whittlesea & L. D. Williams, 1998), people experience a feeling of familiarity when they perceive their processing to be surprising, but for an indefinite reason. This hypothesis has been successful in explaining several illusions of familiarity. Here, it is applied to the prototype-familiarity effect, an illusion of remembering that occurs when people are shown prototype words after studying lists of associates. The experiments showed that studying associates enhances semantic, but not perceptual, processing of prototypes. They also showed that claims of recognizing prototypes can be modified by presenting them in predictive or incongruous contexts at test. The evidence suggests that the effect results from an evaluation process that monitors the coherence of processing. PMID- 11902154 TI - Sugar transport in (hyper)thermophilic archaea. AB - Hyperthermophilic archaea show important metabolic adaptations for growth on carbohydrates under hostile conditions. For carbohydrate uptake so far only ABC type transporters have been described that are equipped with a uniquely high affinity as compared to mesophilic bacterial systems. This allows these organisms to efficiently scavenge all available carbohydrates from the extreme environment. PMID- 11902155 TI - Study of Corynebacterium diphtheriae strains isolated in Romania, northwestern Russia and the Republic of Moldova. AB - A selection of 167 Corynebacterium diphtheriae strains isolated in Romania, the Russian Federation and the Republic of Moldova were analysed by biotyping, phage typing, the toxin production test and by molecular techniques such as ribotyping, pulsed field gel electrophoresis and random amplified polymorphic DNA, in order to establish the epidemiological relatedness, genetic divergence and strain circulation within and between the bordering countries. Using a set of five digoxigenin-labeled oligonucleotides and BstEII digestion, 34 ribotypes were identified. The strains isolated in the epidemic areas (Russia and Moldova) were very closely related but different from those isolated in Romania. C1 and C5 were the main ribotypes identified in these areas. Neither ribotype was found in Romania, where the main circulating types were C3 and C7. Field inversion gel electrophoresis was more discriminative than ribotyping and revealed 54 macrorestriction profiles after SfiI restriction. Both methods showed a significant homogeneity of the strains from epidemic areas and a large diversity among the Romanian strains. Random amplification was useful as an identification method for the epidemic strains, but not for the Romanian ones which displayed a large number of amplification profiles. The phenotypic methods associated with molecular typing techniques enabled distinguishing between strains, detecting the epidemic clone, and sustaining the absence of transmission across borders. PMID- 11902157 TI - Abstracts of the 22nd Annual Conference on Dialysis. Tampa, Florida, USA. March 4 6, 2002. PMID- 11902156 TI - Cholesterol modulates amiodarone-membrane interactions in model and native membranes. AB - The effects of cholesterol, a lipid mostly found in the sarcolemmal membranes, on the interaction of amiodarone with synthetic models of dimyristoylphosphatidylcholine (DMPC) and with native models of mitochondria and brain microsomes was studied. Alterations on the structural order of lipids were assessed by fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) probing the bilayer core, and of the propionic acid derivative 3-(p-(6-phenyl) 1,3,5-hexatrienyl)phenylpropionic acid (DPH-PA) probing the outer regions of the bilayer. As detected by the probes and according to classic observations, cholesterol progressively increased the molecular order in the fluid phase of DMPC. Additionally, it modulated the type and extension of amiodarone effects. For low cholesterol concentrations (< or =10-15 mol%), amiodarone (50 microM) ordered DMPC bilayers and the effects were almost identical to those observed in pure DMPC. For higher cholesterol concentrations, amiodarone ordering effects decreased slightly and faded for cholesterol concentrations as high as 25 and 30 mol%, when detected by DPH-PA and DPH, respectively. Above these high cholesterol concentrations, a crossover from ordering to disordering effects of amiodarone was apparent, either in the upper region of the bilayer or the hydrophobic core. The effects of amiodarone in native membranes of mitochondria and brain microsomes, in which "native" cholesterol accounts for about 0 and 25 mol%, respectively, correlated reasonably with the results in models of synthetic lipids. There is a close relationship between cholesterol concentration and amiodarone effects, in either synthetic models or native model membranes. Therefore, it may be predicted that the lipid physicochemical properties regulated by cholesterol concentration will also modulate the effects of amiodarone in sarcolemma. PMID- 11902158 TI - Humic acid enhanced remediation of an emplaced diesel source in groundwater. 1. Laboratory-based pilot scale test. AB - The enhanced solubility of petroleum-derived compounds in humic acid solutions is the basis for a new groundwater remediation technology. In this unique pilot scale test, a stationary contaminant source consisting of diesel fuel was placed below the water table in a model sand aquifer (1.2 x 5.5 x 1.8-m deep) and flushed with water at a flow rate of 2 cm/h over 5 years. At 51 days, laboratory grade humic acid was added to the water and maintained at a level of approximately 0.8 g/l. The addition of humic acid had only a small impact on the aqueous transport of the BTEX components, which were rapidly dissolved from the diesel, but had a large effect on the flushing of PAHs, including methylated naphthalenes (MNs). Binding to aqueous humic acid enhanced the solubilization of MNs two- to tenfold. During aqueous transport, biodegradation of the BTEX and PAHs occurred, limiting the lateral and longitudinal extent of the diesel contaminant plume in the model aquifer. It appears that through enhanced solubilization, the overall biodegradation rate of the MNs was increased. As the various MNs were depleted from the diesel source, the MN plume shrank and then disappeared. PMID- 11902160 TI - Abstracts of the 4th Annual Meeting of the Working Group on Echocardiography of the European Society of Cardiology. Lisbon, Portugal, 6-9 December 2000. PMID- 11902159 TI - Humic acid enhanced remediation of an emplaced diesel source in groundwater. 2. Numerical model development and application. AB - A pilot scale experiment for humic acid-enhanced remediation of diesel fuel, described in Part 1 of this series, is numerically simulated in three dimensions. Groundwater flow, enhanced solubilization of the diesel source, and reactive transport of the dissolved contaminants and humic acid carrier are solved with a finite element Galerkin approach. The model (BIONAPL) is calibrated by comparing observed and simulated concentrations of seven diesel fuel components (BTEX and methyl-, dimethyl- and trimethylnaphthalene) over a 1500-day monitoring period. Data from supporting bench scale tests were used to estimate contaminant-carrier binding coefficients and to simulate two-site sorption of the carrier to the aquifer sand. The model accurately reproduced the humic acid-induced 10-fold increase in apparent solubility of trimethylnaphthalene. Solubility increases on the order of 2-5 were simulated for methylnaphthalene and dimethylnaphthalene, respectively. Under the experimental and simulated conditions, the residual 500 ml diesel source was almost completely dissolved and degraded within 5 years. Without humic acid flushing, the simulations show complete source dissolution would take about six times longer. PMID- 11902161 TI - Abstracts of the International Anesthesia Research Society 76th Clinical and Scientific Congress. San Diego, California, USA. March 16-20, 2002. PMID- 11902163 TI - Abstracts of the 72nd Annual Meeting of the Zoological Society of Japan. October 6-8, 2001. PMID- 11902162 TI - [Abstracts of the 20th Congress of the German Society of Perinatal Medicine. 29 November-1 December 2001. Berlin, Germany]. PMID- 11902164 TI - Abstracts of the 5th Annual Meeting of the Working Group on Echocardiography of the European Society of Cardiology. Nice, France, 5-8 December 2001. PMID- 11902165 TI - Space biology and medicine. PMID- 11902166 TI - Biological investigations aboard biosatellite Cosmos-782. AB - The Cosmos-782 flight from 25 November to 15 December 1975, carried biological experiments designed to study the effects of weightlessness on insects and fish and on gravitropism and growth in several seed varieties. Investigations carried out on Drosophila melanogaster measured the frequency of recessive lethal mutations and the change in genetic distances in the sex chromosome. The study of Fundulus heteroclitus eggs and fry compared the effects of weightlessness and artificial gravity. Plants experiments studied spatial orientation of over and underground organs of Pinus silvestris and Crepis capillaris seeds. Other investigations used Phycomyces blakesleanus to compare spatial orientation and growth and development in weightlessness and artificial gravity. PMID- 11902167 TI - Engineering and simulation of life sciences Spacelab experiments. AB - The third in a series of Spacelab Mission Development tests was conducted at the Johnson (correction of Johnston) Space Center as a part of the development of Life Sciences experiments for the Space Shuttle era. The latest test was a joint effort of the Ames Research and Johnson Space Centers and utilized animals and men for study. The basic objective of this test was to evaluate the operational concepts planned for the Space Shuttle life science payloads program. A three-man crew (Mission Specialist and two Payload Specialists) conducted 26 experiments and 12 operational tests, which were selected for this 7-day mission simulation. The crew lived on board a simulated Orbiter/Spacelab mockup 24 hr a day. The Orbiter section contained the mid deck crew quarters area, complete with sleeping, galley and waste management provisions. The Spacelab was identical in geometry to the European Space Agency Spacelab design, complete with removable rack sections and stowage provisions. Communications between the crewmen and support personnel were configured and controlled as currently planned for operational shuttle flights. For this test a Science Operations Remote Center was manned at the Ames Research Center and was managed by simulated Mission Control and Payload Operation Control Centers at the Johnson Space Center. This paper presents the test objectives, description of the facilities and test program, and the results of this test. PMID- 11902168 TI - Simulation of space cabin atmosphere. AB - The Soyuz 22 space cabin atmosphere was studied for volatile organic trace contaminants. By gas chromatography the following constituents were identified: methane, ethane, heptane, methanol, ethanol, n-propanol, acetaldehyde, acetone, ethyl benzene. Except for acetone, concentrations of the above compounds were close to the values determined in the mock-up experiments. PMID- 11902169 TI - Radiation danger in prolonged manned space flights. AB - The author examines methods for prognosticating solar activity for several decades to determine optimum periods for prolonged space flight. The focus of the discussion is the presence of a change in magnetic polarity at the beginning of each 11-year solar cycle, resulting in a 22-year cycle. An historical review of solar cycles determined a super cycle of about 180 years. Using this data, it is determined that solar activity will be weak through the end of the 20th century. PMID- 11902170 TI - Space motion sickness. AB - Space motion sickness, presumably triggered by sudden entry into a weightless environment, occurred with unexpected frequency and severity among astronauts who flew the Skylab missions. Recovery from symptoms was complete within 3-5 days, and as revealed by the Skylab M131 Human Vestibular Function Experiment, all crewmembers were immune to experimentally induced motion sickness after mission day 8. This syndrome has been recognized as a possible threat to the early mission well-being and operational efficiency of at least some individuals who will fly space missions in the future. The causes of space motion sickness are not clearly understood, nor have satisfactory methods been identified to date for its prediction, prevention and treatment. In order to minimize the potential impact of this syndrome on Space Shuttle crew operations the National Aeronautics and Space Administration has organized a broad program of inter-disciplinary research involving a large number of scientists in the United States. Current research on the etiology of space motion sickness is based to a large extent on the so called sensory conflict theory. Investigations of the behavioral and neurophysiological consequences of intralabyrinthine, as well as intermodality sensory conflict are being performed. The work in this area is being influenced by the presumed alterations that occur in otolith behavior in weightlessness. In addition to sensory conflict, the possible relationship between observed cephalad shifts of body fluids in weightlessness and space motion sickness is being investigated. Research to date has failed to support the fluid shift theory. Research underway to identify reliable test methods for the prediction of susceptibility to space motion sickness on an individual basis includes attempts to (a) correlate susceptibility in different provocative environments; (b) correlate susceptibility with vestibular and non-vestibular response parameters, the latter including behavioral, hemodynamic and biochemical factors and (c) correlate susceptibility with rate of acquisition and length of retention of sensory adaptation. Controlled studies are also being performed during parabolic flight as a means of attempting to validate predictive tests for susceptibility to this syndrome. Research to develop new or improved countermeasures for space motion sickness is underway in two primary areas. One of these involves anti motion sickness drugs. Significant achievements have been realized with regard to the identification of new highly efficacious drug combinations, dose levels and routes of administration. Although pronounced individual variations must be accounted for in selecting the optimum drug and dose level, combinations of promethazine plus ephedrine or scopolamine plus dexidrine are presently the drugs of choice. Work is also underway to identify side effects associated with anti motion sickness drug use and to identify new drugs which may selectively modify activity in central neural pathways involved in motion sickness. In addition to research on drugs, efforts are being made to develop practical vestibular training methods. Variables which influence rate of acquisition of adaptation, length of retention of adaptation and transfer of protective adaptation to new environments are being evaluated. Also, included in this area is the use of biofeedback and autogenic therapy to train individuals to regulate autonomic responses associated with motion sickness. While valuable new knowledge is expected to evolve from these combined research programs, it is concluded that the final validation of predictive tests and countermeasures will require a series of controlled space flight experiments. PMID- 11902172 TI - A mathematical and experimental simulation of the hematological response to weightlessness. AB - Two ground-based methods of weightlessness simulation--a computer model of erythropoiesis feedback regulation and bedrest--were used to investigate the mechanisms which lead to loss of red cell mass during spaceflight. Both methods were used to simulate the first Skylab mission of 28 days. Human bedrest subjects lose red cell mass linearly with time and in this study the loss was 6.7% at the end of four weeks (compared to 14% in Skylab). Postbedrest recovery of red cell mass was delayed for two weeks during which time a further decline in this quantity was noted. This is consistent with the first Skylab mission but not with the two longer flights of two and three months. Hemoconcentration, observed early in the study, was essentially maintained despite red cell loss because of continued loss of plasma volume. The computer model, using the time-varying hematocrit data to estimate red cell production rates, predicted dynamic behavior of plasma volume and red cell mass that was in close agreement with the measured values. The results support the hypothesis that red cell loss during supine bedrest is a normal physiological feedback process in response to hemoconcentration enhanced tissue oxygenation and suppression of red cell production. In contrast, the delayed postbedrest recovery of red cell mass was more difficult to explain, especially in the light of enhanced reticulocyte indices observed at the onset on ambulation. Model simulation suggested the possibilities, still to be experimentally demonstrated, that this period was marked by some combination of increased oxygen-hemoglobin affinity, small reductions in mean red cell life span, ineffective erythropoiesis, or abnormal reticulocytosis. The question of whether hemoconcentration is the sole contributor to spaceflight red cell losses also remains to be resolved. PMID- 11902171 TI - Exercise response to simulated weightlessness. AB - Two bed rest analog studies of space flight were performed; one 14 d and the other 28 d in duration. Exercise response was studied in detail during the 28 d study and following both the 14 d and 28 d studies. This paper relates the results of these studies to physiologic changes noted during and following space flight. The most consistent change noted after both bed rest and space flight is an elevated heart rate during exercise. A second consistent finding is a postflight or postbed rest reduction in cardiac stroke volume. Cardiac output changes were variable. The inability to simulate inflight activity levels and personal exercise makes a direct comparison between bed rest and the results from specific space flights difficult. PMID- 11902173 TI - Effects of prolonged space flight on rat skeletal muscle. AB - The effect of a 20-day space flight on water, Na+, K+, Mg2+, Ca2+ and glycogen contents as well as on activities of glycogen metabolism enzymes--glycogen synthetase and glycogen phosphorylase--of rat skeletal muscles was studied. This data is regarded as an integral test characterizing the state of contractile tissue of the animals at the final stage of flight aboard biosatellites. The measurements indicate that there were no significant changes of cations and glycogen contents nor of the enzymic activities in fast-twitch muscles during the 20-day spaceflight. At the same time dehydration in these muscles was observed, which disappeared on the 25th postflight day. In slow-twitch antigravitational skeletal muscle (m. soleus) there was a decrease of K+ and increase of Na+ in the tissue contents. The changes disappeared at the end of the on-earth readaptation period. From the pattern of these observations, we can conclude that the 20-day space flight leads to some reversible biochemical changes of the rat skeletal muscles. A conclusion can be drawn about necessity of creating, aboard the spaceship, an artificial load on antigravitational skeletal muscles. PMID- 11902174 TI - A study of metabolic balance in crewmembers of Skylab IV. AB - A metabolic balance study was conducted on the three crewmembers of the 84-day Skylab IV earth orbital mission. Dietary intake was controlled, monitored, and kept very nearly constant for a period commencing 21 days prior to flight, throughout flight, and for a period of 18 days postflight. Within the first 30 days of flight urine calcium rose to a level approx. 100% above preflight levels and remained elevated for the remainder of the flight. Fecal calcium excretion increased more slowly but continued to accelerate throughout the flight and did not return to baseline levels during the postflight period. Urinary nitrogen increased to 25-30% above preflight levels within one month following launch and thereafter gradually subsided toward control values. The overall losses of calcium averaged approx. 200 mg per day throughout the mission while nitrogen losses averaged 590 mg. Various other indices of musculoskeletal deterioration are discussed and correlated. The parallelism between the effects of weightlessness and bed rest is reviewed. It is noted, that no evidence is yet available as to the identity of the initial biological response to the absence of gravity. PMID- 11902175 TI - Amino aciduria in weightlessness. AB - Urinary excretion of amino acids by the 9 Skylab crewmen was studied as an indicator of the metabolic effects caused by exposure to the space flight environment. Intake was consistent in quality and quantity throughout the 28, 59 and 84-day flights for each of the crewmen and complete collections were accomplished. The results indicated an increased excretion in most amino acids during the first month of flight which remained elevated in the second and third months but to a lesser extent. Additional indications of change in muscle and skeletal metabolism were observed. These results point to the desirability of obtaining additional indices of alterations in protein synthetic processes in conjunction with future space flights. PMID- 11902176 TI - Fluid volumes changes induced by spaceflight. AB - The blood volume (BV), plasma volume (PV), and extracellular fluid volume changes produced in crewmembers during spaceflights of 11-84 days were compared to changes after 14 or 28 days of bedrest. Spaceflight and bedrest produce approximately equal BV changes but the recorded PV change after spaceflight was less. However, the diurnal change in PV may explain the smaller decreases recorded after spaceflight. The cardiovascular deconditioning caused by spaceflight and bedrest was compared using the mean heart rate response to lower body negative pressure (LBNP) testing at -50 mmHg pressure. These tests show approximately equal LBNP produced heart rate changes after bedrest and spaceflight. A countermeasure which includes 4 hr of LBNP treatment at -30 mmHg and the ingestion of one l. of saline was studied and found capable of returning the heart rate response and the PV of bedrested subjects to control (prebedrest) levels suggesting that it would be useful to the crewmembers after a spaceflight. PMID- 11902177 TI - Lunar polar ice deposits: scientific and utilization objectives of the Lunar Ice Discovery Mission proposal. AB - The Clementine mission has revived interest in the possibility that ice exists in shadowed craters near the lunar poles. Theoretically, the problem is complex, with several possible sources of water (meteoroid, asteroid, comet impact), several possible loss mechanisms (impact vaporization, sputtering, photoionization), and burial by meteorite impact. Opinions of modelers have ranged from no ice to several times 10(16) g of ice in the cold traps. Clementine bistatic radar data have been interpreted in favor of the presence of ice, while Arecibo radar data do not confirm its presence. The Lunar Prospector mission, planned to be flown in the fall of 1997, could gather new evidence for the existence of ice. If ice is present, both scientific and utilitarian objectives would be addressed by a lunar polar rover, such as that proposed to the NASA Discovery program, but not selected. The lunar polar rover remains the best way to understand the distribution and characteristics of lunar polar ice. PMID- 11902179 TI - So-called "partial birth abortion" bans: bad medicine? Maybe. Bad law? Definitely! PMID- 11902178 TI - Early renal changes in 45 degrees HDT rats. AB - BACKGROUND: Both microgravity and simulated microgravity models, such as the 45HDT (45 degrees head-down tilt), cause a redistribution of body fluids indicating a possible adaptive process to the microgravity stressor. Understanding the physiological processes that occur in microgravity is a first step to developing countermeasures to stop its harmful effects, i.e., (edema, motion sickness) during long-term space flights. HYPOTHESIS: Because of the kidneys' functional role in the regulation of fluid volume in the body, it plays a key role in the body's adaptation to microgravity. METHODS: Rats were injected intramuscularly with a radioactive tracer and then lightly anesthetized in order to facilitate their placement in the 45HDT position. They were then placed in the 45HDT position using a specially designed ramp (45HDT group) or prone position (control group) for an experimental time period of 1 h. During this period, the 99mTc-DTPA (technetium-labeled diethylenepentaacetate, MW=492 amu, physical half life of 6.02 h) radioactive tracer clearance rate was determined by measuring gamma counts per minute. The kidneys were then fixed and sectioned for electron microscopy. A point counting method was used to quantitate intracellular spaces of the kidney proximal tubules. RESULTS: 45HDT animals show a significantly (p=0.0001) increased area in the interstitial space of the proximal tubules. CONCLUSIONS: There are significant changes in the kidneys during a 1 h exposure to a simulated microgravity environment that consist primarily of anatomical alterations in the kidney proximal tubules. The kidneys also appear to respond differently to the initial periods of head-down tilt. PMID- 11902180 TI - Research misconduct--an indictment and possible solution. AB - The core problems of scientific misconduct are systemic to the infrastructure of the conduct of scientific research itself, and therefore are probably immune to any short-term solutions. Renaming federal agencies, reorganizing monitoring activities, appointing new personnel, and other similar proposed remedies are necessary but insufficient measures. Indeed, the etiological factors in misconduct include an erosion of trust that stands in the way of the ethical integrity flowing from the major ethical principles identified in the Belmont Report. To make those principles applicable and useful to meet the challenges of current research activities, especially those involving human subjects, depends on our ability to foster an unprecedented cooperation of community resources. PMID- 11902181 TI - Trials with errors--preserving the integrity of clinical trials. AB - The crucial final test of medical research is the clinical trial, which determines whether a drug or discovery really is an effective therapy. All people who participate in clinical trials--researchers, sponsors, volunteers, analysts, reviewers, overseers, others--have opportunities to strengthen or weaken the integrity of the trial system by their behavior. Medical research is now officially married to business, and "profitable" connotes something different to each partner. Only if research and business can profit in parallel will the alliance succeed. Every person who is involved in the medical research business faces temptations and must choose how to react. Each has power and must choose how to wield it. Several centuries before this marriage, the Englishman Izaak Walton noted that "Health is...a blessing that money cannot buy." PMID- 11902182 TI - Protecting the human subjects of social science research--the role of institutional review boards. AB - Current and proposed reforms of the regulatory schema for protecting human subjects of research have focused attention on Institutional Review Board (IRB) responsibilities. Consensus on the need for strengthening the oversight of these boards is all but certain--with the exception of research in the social and behavioral sciences, where an argument for less oversight is being made. The thesis in this article is that respecting the salient features of research in the social and behavioral sciences will ameliorate the tensions leading to this demand and offer better protection to the subjects of social and behavior studies. PMID- 11902183 TI - Protecting human subjects in research--occasional views along a road less traveled. AB - Public trust in biomedical research is eroding rapidly because too many investigators participating in human subjects research have failed to take personal responsibility for their actions. In this essay, taken partly from an address to the 66th Annual Research Colloquium of the Massachusetts General Hospital, January 2001, and a presentation to the research community of the Washington University School of Medicine in September, 2000, in St. Louis, Missouri; Greg Koski shares his views about protecting human subjects in biomedical research. PMID- 11902184 TI - Clinical research involving children. PMID- 11902185 TI - Taking the "I" out of IRB--and putting "community" in. AB - If one looks back on the history of American research ethics, a bold pattern emerges. Since World War II, about every twenty years or so a breach of the social contract between investigators and human research subjects galvanizes public and professional interest in the ethical foundations and oversight mechanisms governing research with humans. PMID- 11902186 TI - Brave new egg. PMID- 11902187 TI - On the disposition of frozen embryos. PMID- 11902191 TI - Rationality and principles: a criticism of the ethic of care. PMID- 11902192 TI - The ethical implications of the Human Genome Project for the workplace. PMID- 11902193 TI - The ethics of the broader usage of Prozac: social choice or social bias? PMID- 11902194 TI - The relationship between ethical ideology and ethical behavior intentions: an exploratory look at physicians' responses to managed care dilemmas. AB - Within the past few years, managed care health insurance programs have become commonplace. With managed care programs, however, physicians are facing increasing ethical pressures. This paper examines the relationship between physicians' behavior intentions with respect to four managed care ethical scenarios and their responses to Forsyth's (1980) Ethics Position Questionnaire (EPQ). This is one of the first papers to compare this scale to behavioral intentions in the workplace. We provide a literature review of the ethical dilemmas that doctors face under a managed care system and conduct a national random sample of general practitioners and surgeons regarding the four managed care ethical dilemmas. The results show that the doctors surveyed are significantly more idealistic than relativistic. In relating the EPQ to the ethical scenarios, however, there was no support for the proposition that ethical ideology was related to the ethical behavioral intentions. This suggests more research is needed to establish the links between ethical positions, attitudes, and behavioral intentions. Finally, there were little differences in EPQ scores by practice or demographic variables, the only significant result being that general surgeons are significantly more idealistic than family practitioners. PMID- 11902195 TI - Treatment with azithromycin improves endothelial function. PMID- 11902196 TI - Clot-busters may increase mortality in the elderly. PMID- 11902197 TI - One previous owner. PMID- 11902198 TI - In memoriam: tougher rules could be the legacy of gene therapy's first death. PMID- 11902199 TI - Health and happiness...or doom and gloom? PMID- 11902200 TI - Before their time. Surgery on the unborn is a reality. But can it be justified when a baby's life is not at risk? PMID- 11902201 TI - Genetic bounty. PMID- 11902202 TI - The green man: invoking God and nature won't solve our problems with biotechnology. PMID- 11902205 TI - Gene warrior. Interview by Ehsan Masood. PMID- 11902203 TI - Looking for a miracle. PMID- 11902210 TI - Birth control as an international program. PMID- 11902211 TI - Assisted reproduction and reproductive rights: the case of in vitro fertilization. AB - In vitro fertilization (IVF) and other assisted reproduction technologies (ARTs) have become widely accepted as therapy for a wide array of fertility problems and accompanied by the rapid expansion of clinics that provide full range of ARTs. Although these technologies undoubtedly offer benefits for some individuals, they raise important questions over reproductive rights to safe and effective treatment as well as access. This article analyzes current data concerning the safety, effectiveness, and cost of IVF. It concludes that IVF and related techniques have been transformed too rapidly and easily from experimental to therapy status, despite evidence that suggests considerable caution is warranted. Unfortunately, the widespread diffusion of IVF has preceded rather than followed firm evidence of its value in extending the reproductive rights of women and couples. Resources might better be directed toward prevention of fertility problems and discovering the causes of infertility. PMID- 11902212 TI - Coping with Dolly: scenes from the National Bioethics Advisory Commission. PMID- 11902213 TI - Creating a clone in ninety days: in search of a cloning policy. PMID- 11902214 TI - Report to the European Commission: ethical aspects of cloning techniques. PMID- 11902215 TI - The crucial role of stewardship in health care ethics. PMID- 11902216 TI - A case of misdirected love? In vitro fertilization the the quest for fertility. PMID- 11902217 TI - Collective action and the goals of medicine. PMID- 11902218 TI - COX-2 inhibitors: a role in Alzheimer's disease? PMID- 11902219 TI - Lotronex(tm): therapy for diarrhea predominant irritable bowel syndrome. PMID- 11902220 TI - Entacapone: adjunctive use in patients with advanced Parkinson's disease. PMID- 11902221 TI - Tenecteplase: single-bolus thrombolytic therapy for acute myocardial infarction. PMID- 11902222 TI - Prevnar(tm): a pneumococcal conjugate vaccine for infants and young children. PMID- 11902223 TI - Linezolid for the treatment of serious gram-positive infections. PMID- 11902224 TI - Vagus nerve stimulation (VNS) for treatment-resistant depression. PMID- 11902225 TI - Transdermal contraceptive patch--a new birth control option. PMID- 11902226 TI - Do neuraminidase inhibitors prevent influenza? PMID- 11902227 TI - Preventing restraint deaths. PMID- 11902228 TI - Infant abductions: preventing future occurrences. PMID- 11902229 TI - Blood transfusion errors: preventing future occurrences. PMID- 11902230 TI - High-alert medications and patient safety. PMID- 11902231 TI - Operative and post-operative complications: lessons for the future. PMID- 11902232 TI - Making an impact on health care. PMID- 11902233 TI - Fatal falls: lessons for the future. AB - Health care organizations that have experienced sentinel events due to falls have identified the root causes and risk reduction strategies included in this issue. In addition, experts have commented on the events and the related root causes and risk reduction strategies. The Joint Commission offers this information for consideration by hospitals, long term care facilities and behavioral health care organizations in their continuing efforts to reduce the risk of falls of their patients, residents or individuals served. PMID- 11902234 TI - Infusion pumps: preventing future adverse events. AB - The Joint Commission offers this information for health care organizations to consider in their continuing efforts to reduce the risk of adverse events in the care of their patients, residents or individuals served. In the past two years, health care organizations have reported to JCAHO two sentinel events involving the use of infusion pumps. While it is difficult to reach meaningful conclusions from only two cases, these cases have raised a concern about patient safety involving the use of infusion pumps. In recent months there has been a great deal of discussion about potential problems with infusion pumps. Therefore, JCAHO provides the following expert comments and recommendations to alert health care organizations on how to prevent such adverse events from occurring. PMID- 11902235 TI - Mix-up leads to a medication error. AB - Recently, the Joint Commission's Board of Commissioners approved new and revised standards directly focused on patient safety and medical/health care error reduction in hospitals. The standards, to be implemented July 1, 2001, augment the nearly 50 percent of current Joint Commission standards directly related to patient safety. This issue of Sentinel Event Alert includes information accredited hospitals may use in conducting a proactive risk assessment, a requirement of the new patient safety standards. PMID- 11902236 TI - Lessons learned: fires in the home care setting. PMID- 11902237 TI - Kernicterus threatens healthy newborns. PMID- 11902239 TI - Exposure to Creutzfeldt-Jakob Disease. PMID- 11902238 TI - Look-alike, sound-alike drug names. PMID- 11902240 TI - Medical gas mix-ups. PMID- 11902241 TI - Preventing needlestick and sharps injuries. PMID- 11902242 TI - Medication errors related to potentially dangerous abbreviations. PMID- 11902243 TI - A follow-up review of wrong site surgery. PMID- 11902244 TI - Preventing ventilator-related deaths and injuries. PMID- 11902245 TI - Evidence report finds corticosteroids benefit COPD exacerbations. PMID- 11902246 TI - Report reviews stroke intervention strategies. PMID- 11902247 TI - Guidelines on high blood pressure in pregnancy give new classifications of disease. PMID- 11902248 TI - Guidelines on diabetic foot disorders stress foot exams, multidisciplinary management as key preventive measures. PMID- 11902249 TI - Randomized, placebo-controlled trial of lisofylline for early treatment of acute lung injury and acute respiratory distress syndrome. AB - OBJECTIVE: To determine whether the administration of lisofylline (1-[5R hydroxyhexyl]-3,7-dimethylxanthine) would decrease mortality in patients with acute lung injury (ALI) or acute respiratory distress syndrome (ARDS). DESIGN: A prospective, randomized, double-blind, placebo-controlled, multicenter study. SETTING: Intensive care units at 21 hospitals at the ten centers constituting the ARDS Clinical Trials Network. PATIENTS: A total of 235 patients who met eligibility criteria were enrolled in the study (116 into the lisofylline group, 119 into the placebo group). INTERVENTIONS: Patients were randomized to receive either lisofylline or placebo. The dose of lisofylline was 3 mg/kg with a maximum dose of 300 mg intravenously every 6 hrs. The intravenous solution of study drug was administered over 10 mins every 6 hrs. Dosing was continued for 20 days or until the patient achieved 48 hrs of unassisted breathing. MEASUREMENTS AND MAIN RESULTS: The trial was stopped by the Data Safety Monitoring Board for futility at the first scheduled interim analysis. The patient groups had similar characteristics at enrollment. No significant safety concerns were associated with lisofylline therapy. There was no significant difference between groups in the number of patients who had died at 28 days (31.9% lisofylline vs. 24.7% placebo, p = .215). There was no significant difference between the lisofylline and placebo groups in terms of resolution of organ failures, ventilator-free days, infection-related deaths, or development of serious infection during the 28 day study period. The median number of organ failure-free days for the five nonpulmonary organ failures examined (cardiovascular, central nervous system, coagulation, hepatic, and renal) was not different between the lisofylline and placebo groups. Although lisofylline has been reported to decrease circulating free fatty acid levels, we did not find any such treatment effect compared with placebo. CONCLUSIONS: In this study, there was no evidence that lisofylline had beneficial effects in the treatment of established ALI/ARDS. PMID- 11902250 TI - A phase II randomized, controlled trial of continuous hemofiltration in sepsis. AB - OBJECTIVE: To study the effect of early and continuous venovenous hemofiltration (CVVH) on the plasma concentrations of several humoral mediators of inflammation and subsequent organ dysfunction in septic patients. DESIGN: Randomized, controlled trial. SETTING: Intensive care unit of a tertiary hospital. PATIENTS: Twenty-four patients with early septic shock or septic organ dysfunction. INTERVENTIONS: Random allocation to receive 48 hrs of isovolemic CVVH at 2 L/hr of fluid exchange or no hemofiltration. MEASUREMENTS AND MAIN RESULTS: We measured the plasma concentrations of complement fractions C3a and C5a, interleukins 6, 8, and 10, and tumor necrosis factor alpha at baseline and 2, 24, 26, 48, and 72 hrs. A multiple organ dysfunction score (MODS) was calculated daily for each patient until death or discharge from the intensive care unit. The concentrations of most mediators decreased between baseline and 72 hrs. Some significant falls in concentration could be identified between specific time points, but CVVH was not associated with an overall reduction in any plasma cytokine concentrations. There was also no difference between the mean cumulative MODS for control survivors (43.3 +/- 19.7) and CVVH survivors (33.2 +/- 19.0; p = .30), and no difference between the average MODS calculated for all controls (4.1 +/- 1.9) and all CVVH subjects (3.3 +/- 1.7; p = .26). CVVH did not improve oxygenation, lower the platelet count, or reduce the duration of vasopressor support and mechanical ventilation. CONCLUSIONS: Early use of CVVH at 2 L/hr did not reduce the circulating concentrations of several cytokines and anaphylatoxins associated with septic shock, or the organ dysfunction that followed severe sepsis. CVVH using current technology cannot be recommended as an adjunct to the treatment of septic shock unless severe acute renal failure is present. PMID- 11902251 TI - Low-dose prostacyclin preserves renal function in high-risk patients after coronary bypass surgery. AB - OBJECTIVE: Renal failure after bypass is still a threatening problem prolonging hospital care and reducing overall survival. The following pilot study was aimed to analyze whether perioperative low-dose prostacyclin infusion is able to preserve renal function in a selected group of patients who according to a poor cardiac function were stratified as high risk for the development of renal failure after bypass. DESIGN: Prospective randomized study. SETTING: Tertiary care university medical center. PATIENTS: Thirty-four patients scheduled for primary cardiac bypass surgery were included in the study (prostacyclin n = 17, control n = 17). Inclusion criteria were normal renal function before surgery and a cardiac ejection fraction <40%. INTERVENTIONS: Low-dose prostacyclin (2 ng/kg/min) was added to the standard anesthetic protocol. Infusion was started immediately before surgery and was continued for a maximum of 48 MEASUREMENTS AND MAIN RESULTS: Significant differences in the endogenous creatinine clearance were found between the prostacyclin and the control group. Whereas there was a significant drop in the creatinine clearance at 6 hrs after surgery in the control group with a prolonged recovery period, values in the prostacyclin group remained stable. Creatinine clearance before intervention was 100 +/- 22 mL/min in the control group and 91 +/- 22 mL/min in the prostacyclin group, values at 24 hr were 68 +/- 34 mL/min vs. 103 +/- 37 mL/min, respectively (p < .01). Significant findings in favor for the prostacyclin group were also found for urine output and the fractional excretion rate of sodium. CONCLUSION: This first pilot study indicates that low-dose prostacyclin may be of substantial value for preserving renal function in high-risk patients after coronary bypass surgery. PMID- 11902252 TI - Sedation, analgesia, and neuromuscular blockade of the critically ill adult: revised clinical practice guidelines for 2002. PMID- 11902253 TI - Clinical practice guidelines for the sustained use of sedatives and analgesics in the critically ill adult. PMID- 11902254 TI - Cardiomyocyte intracellular calcium and cardiac dysfunction after burn trauma. AB - OBJECTIVE: To examine the effects of pharmacologic agents designed to limit burn mediated calcium overload on cardiomyocyte [Ca2+] and cardiac contractile function. DESIGN: Experimental, comparative study. SETTING: Cellular biology and physiology laboratory. SUBJECTS: Adult Sprague Dawley rats. INTERVENTIONS: Rats were given third-degree burn injury over 40% of the total body surface area, were fluid resuscitated, and then were divided randomly to receive one of five treatments: vehicle (normal saline); amiloride (50 mg/kg) to inhibit H+-Na+ exchange and subsequent Na+-Ca2+ exchange; dantrolene (10 mg/kg, 30 mins, 6 and 22 hrs postburn) to inhibit sarcoplasmic reticulum Ca2+ release; diltiazem (10 mg/kg given over first 6 hrs postburn); or amlodipine (0.07 mg/kg, 24 hrs preburn and 30 mins postburn) to block calcium slow channels. Appropriate controls (sham burns given the appropriate pharmacologic agent) were included in each group. Twenty-four hrs postburn, left ventricular function (Langendorff), cardiomyocyte [Ca2+]i and [Na+]i measured by fura-2-AM or sodium-binding benzofurzan isophthalate loading of cardiomyocytes, and myocyte secretion of tumor necrosis factor-alpha (enzyme-linked immunosorbent assay) were assessed in shams and burns from each experimental group. This time point was selected based on our previous work confirming maximal ventricular contractile defects and maximal cytokine secretion 24 hrs postburn. MEASUREMENTS AND MAIN RESULTS: Burn trauma increased myocyte [Ca2+]i and [Na+]i, promoted tumor necrosis factor-alpha secretion by cardiomyocytes, and impaired left ventricular function. All pharmacologic agents reduced the burn-mediated Ca2+/Na+ accumulation in cardiomyocytes and ablated burn-mediated tumor necrosis factor-alpha secretion by myocytes; in contrast, dantrolene and amiloride provided significantly greater cardioprotection than pharmacologic agents that specifically targeted Ca2+ slow channels (diltiazem and amlodipine). CONCLUSION: Our data suggest that the calcium antagonists used in this study provide cardioprotection by modulating several aspects of the overall inflammatory cascade rather than solely limiting cardiomyocyte accumulation of calcium. PMID- 11902255 TI - Clinical practice guidelines for sustained neuromuscular blockade in the adult critically ill patient. PMID- 11902257 TI - Marked difference in pathophysiology between tissue factor- and lipopolysaccharide-induced disseminated intravascular coagulation models in rats. AB - OBJECTIVE: Tissue factor and lipopolysaccharide frequently have been used to induce disseminated intravascular coagulation in experimental animal models. Although the pathophysiology of disseminated intravascular coagulation may differ according to the agents used to induce it, these previous models have not distinguished between the use of different disseminated intravascular coagulation inducing agents. In this study, we attempted to evaluate the characteristic features of these agents in two types of disseminated intravascular coagulation models, with special reference to selected hemostatic parameters and pathologic findings in the kidney. DESIGN: Prospective, comparative, experimental study. SETTING: Laboratory at a university hospital. SUBJECTS: Twenty-seven male Wistar rats, age 6-7 wks, weighing 160-170 g. INTERVENTIONS: Three groups of animals were studied: a control group (n = 8) receiving physiologic saline, a tissue factor-treated group (n = 11) receiving tissue factor 3.75 units/kg, and a lipopolysaccharide-treated group (n = 8) receiving lipopolysaccharide 30 mg/kg; each group sustained infusion for 4 hrs via the tail vein. MEASUREMENTS AND MAIN RESULTS: The degree of hemostatic activation in both types of experimental disseminated intravascular coagulation was identical, based on the results of thrombin-antithrombin III complex levels. Markedly elevated D-dimer concentrations were observed without organ dysfunction or fibrin deposition in the kidney on administration of tissue factor, whereas markedly elevated plasminogen activator inhibitor activity, decreased antithrombin III activity, severe organ failure, and marked fibrin deposition in the kidney were observed for lipopolysaccharide administration. CONCLUSION: Because pathophysiology differed remarkably between the tissue factor- and lipopolysaccharide-induced disseminated intravascular coagulation models in rats, we recommend that they be assessed carefully as distinct entities to determine implications of their experimental and clinical use. PMID- 11902256 TI - Crystalloid strong ion difference determines metabolic acid-base change during in vitro hemodilution. AB - OBJECTIVES: To determine the relationship between the strong ion difference (SID) of a diluting crystalloid and its metabolic acid-base effects on in vitro blood dilution. DESIGN: Prospective in vitro study. SETTING: University research laboratory. SUBJECTS: Normal human blood. INTERVENTIONS: Three solutions were prepared, each with [Na] 140 mmol/L. [Cl-] for solutions 1, 2, and 3 was 120, 110, and 100 mmol/L, respectively, the other anion being HCO3-. SID values were thus 20, 30, and 40 mEq/L, respectively. Serial dilutions of well-oxygenated fresh venous blood were performed anaerobically by using each of solutions 1-3 as well as 0.9% saline (SID = 0 mEq/L) and Hartmann's solution (SID = -4 mEq/L). MEASUREMENTS AND MAIN RESULTS: Blood gas and electrolyte analyses were performed before and after each dilution. Apart from dilutions with solution 3 (crystalloid SID 40 mEq/L) during which plasma SID did not change, plasma SID decreased during hemodilution. In contrast, base excess increased during hemodilution with solutions 3 and 2 (crystalloid SID 40 mEq/L and 30 mEq/L, respectively) and decreased only with the remaining three solutions. The relationships between hemoglobin concentrations and both plasma SID and whole blood base excess throughout dilution were linear, with slopes proportional to the SID of the diluent in each case. Linear regression revealed that the SID of crystalloid producing a zero base excess/hemoglobin concentration slope during blood dilution (i.e., no change in metabolic acid-base status) is 23.7 mEq/L. CONCLUSIONS: On in vitro hemodilution, there is a simple linear relationship between diluent crystalloid SID and the rate and direction of change of plasma SID and whole blood base excess. Direct extrapolation to in vivo situations such as acute normovolemic hemodilution and large volume correction of extracellular fluid deficits requires experimental confirmation. PMID- 11902258 TI - Effects of endotoxin tolerance on liver function after hepatic ischemia/reperfusion injury in the rat. AB - OBJECTIVE: It is known that endotoxin tolerance prevents lethality after ischemia/reperfusion injuries (e.g., myocardial infarction) in laboratory animals. We used a rat model of partial hepatic ischemia/reperfusion to investigate whether endotoxin tolerance prevents associated lethality and disorders of liver function. DESIGN: Prospective animal study. SETTING: University research facility. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Hepatic ischemia was initiated by atraumatic clipping across the portal venous and hepatic arterial blood supply to the left lateral lobe for 90 mins. The common bile duct was canalized, and in a second set of experiments the bile duct of the left lateral lobe was canalized selectively. Bile flow, bile acids, and transaminases were determined during ischemia and 300 mins of reperfusion in endotoxin-tolerant and -nontolerant rats. MEASUREMENTS AND MAIN RESULTS: Endotoxin-nontolerant animals showed a 50% lethality after hepatic ischemia/reperfusion injuries. All endotoxin-tolerant rats survived and did not react with any change in bile flow, showing a constant flow. The amount of bile acids in the common bile duct was reduced during ischemia and regained the concentrations of sham-operated animals 60 mins after reperfusion. From 180 mins after reperfusion, the difference between endotoxin-tolerant and -nontolerant animals was statistically significant. When bile acid concentration was determined in the ischemic left lateral lobe, ischemia/reperfusion was found to significantly decrease in endotoxin-nontolerant rats 60 mins after reperfusion. In contrast, endotoxin-tolerant rats produced normal amounts of bile acids 60 mins after reperfusion. At 120 mins after reperfusion, the amount of bile acids in the formerly ischemic left lateral lobe was more than normal. CONCLUSIONS: In this model of partial hepatic ischemia/reperfusion, endotoxin tolerance prevents ischemia/reperfusion injury-associated lethality and local disorders of liver function. This phenomenon induced by endotoxin tolerance may be useful in liver surgery to prevent ischemia/reperfusion injury. PMID- 11902259 TI - Anti-ovine interleukin-1beta monoclonal antibody immunotherapy in an ovine model of gram-negative septic shock. AB - OBJECTIVE: To investigate the efficacy of an anti-ovine interleukin-1beta monoclonal antibody to ameliorate pathophysiological derangements and improve survival in an ovine model of gram-negative septic shock. DESIGN: Prospective, placebo-controlled, interventional study (24-hr study period). SETTING: University hospital animal research laboratory. SUBJECTS: Ten awake, mature female sheep. INTERVENTIONS: Seven milligrams per kilogram of intravenous anti ovine interleukin-1beta immunoglobin G1 monoclonal antibody (anti-interleukin 1beta group, n = 5) or equivalent amount of protein (5% human albumin; control group, n = 5) was infused over 1 hr (time-zero minus 1 hr to time-zero) and followed by an intravenous LD100 live Escherichia coli infusion (time-zero to time-zero plus 1 hr). Normal saline, maintenance and boluses to maintain baseline filling pressures, and gentamicin, 3 mg/kg intravenous, at time-zero plus 2 and time-zero plus 13 hrs. MEASUREMENTS AND MAIN RESULTS: Hemodynamic and oxygen transport indexes as well as hematological, biochemical, cytokine (interleukin 1beta, tumor necrosis factor-alpha), and endotoxin measurements were performed at baseline (time-zero minus 1 hr), on completion of the monoclonal antibody/placebo (time-zero) and E. coli (time-zero plus 1 hr) infusions, and at multiple time points thereafter (time-zero plus 1.5 hrs to time-zero plus 24 hrs). Baseline data were not different between the treatment groups. From time-zero plus 1.5 hrs onward, in the anti-interleukin-1beta group, there was a sustained increase in mean arterial pressure, decreased peripheral vasodilation, and an attenuated metabolic acidosis, relative to the control group (p < or = .01, repeated measures analysis of variance). Predicted percentage increases in mean arterial pressure and systemic vascular resistance index relative to the control group were 35% and 40%, respectively. Resuscitation fluid requirements were also decreased: anti-interleukin-1beta group, 4.1 +/- 2.9 mL x kg(-1) x hr(-1); control group, 10.6 +/- 1.8 mL x kg(-1) x hr(-1) (p < or = .01, Student's t test). Survival was not different (anti-interleukin-1beta group, 40%; control group, 0%; p > .01, log-rank test). CONCLUSIONS: Adjunctive therapy with anti ovine interleukin-1beta monoclonal antibody in ovine gram-negative septic shock was associated with improved hemodynamic performance. However, the beneficial effects were incomplete and survival was not significantly improved. PMID- 11902260 TI - Partial liquid ventilation and positive end-expiratory pressure reduce ventilator induced lung injury in an ovine model of acute respiratory failure. AB - OBJECTIVE: To examine the isolated and combined effects of positive end expiratory pressure (PEEP) and partial liquid ventilation (PLV) on the development of ventilator-induced lung injury in an ovine model. DESIGN: Prospective controlled animal study. SETTING: University-based cardiovascular animal physiology laboratory. SUBJECTS: Thirty-eight anesthetized supine sheep weighing 22.3 +/- 2.2 kg. INTERVENTIONS: Animals were ventilated for 6 hrs (respiratory rate, 15; FIO2, 1.0, inspiratory/expiratory ratio, 1:1) with one of five pressure-controlled strategies, expressed as peak inspiratory pressure (PIP)/PEEP: low-PIP, 25/5 cm H2O (n = 8); high-PIP, 50/5 cm H2O (n = 8); high-PIP PLV, 50/5 cm H2O-PLV (n = 8); high-PEEP, 50/20 cm H2O (n = 7); and high-PEEP-PLV, 50/20 cm H2O-PLV (n = 7). MEASUREMENTS AND MAIN RESULTS: Compared with the low PIP control, high-PIP ventilation increased airleak, shunt, histologic evidence of lung injury, neutrophil infiltrates, and wet lung weight. Maintaining PEEP at 20 cm H2O or adding PLV reduced the development of physiologic shunt and dependent histologic injury indexes. Neither higher PEEP nor PLV reduced the high incidence of barotrauma observed in high-PIP animals. CONCLUSIONS: We conclude that application of PLV or PEEP at 20 cm H2O may improve gas exchange and afford lung protection from ventilator-induced lung injury during high-pressure mechanical ventilation in this model. PMID- 11902261 TI - Thioredoxin is associated with endotoxin tolerance in mice. AB - OBJECTIVE: Oxidative stress and subsequent lipid peroxidation appear to be central to the lethal effect of lipopolysaccharide. We hypothesized that induction of an antioxidant protein, thioredoxin, would play an important role in the development of endotoxin tolerance and reduce mortality rates in lipopolysaccharide-treated mice. DESIGN: Prospective, randomized, controlled study. SETTING: University research laboratory. SUBJECTS: Adult, male, ddy mice. INTERVENTIONS: In survival-curve experiments, mice were pre-treated intravenously with a low dose of Escherichia coli lipopolysaccharide (20 microg/mouse, pretreatment group) or saline (control group). A large dose of lipopolysaccharide (200 microg/mouse) subsequently was injected into the tail vein 16 hrs after pretreatment. In experiments to measure the expression of thioredoxin after lipopolysaccharide challenge, mice were injected intravenously with different concentrations of lipopolysaccharide (between 20 and 200 microg/mouse, designated the lipopolysaccharide group) or with saline (control group). MEASUREMENTS AND MAIN RESULTS: The survival rate during the 72-hr observation period was significantly higher in the pretreatment group (82%) than in the control group (30%; p = .025 by Mantel-Cox log rank analysis). After lipopolysaccharide challenge, thioredoxin concentrations in the lung, heart, and liver of mice in the pretreated group were about 1.5- to 2-fold higher than those in the control group. Enhancement in these organs became apparent about 6 hrs after lipopolysaccharide challenge and lasted until at least 24 hrs. The levels of accumulation of 4-hydroxy-2-nonenal modified proteins after a large dose of lipopolysaccharide challenge were lower in the pretreatment group than in the control group. CONCLUSIONS: These results demonstrate that mice with an increased concentration of thioredoxin, induced by pretreatment with a low dose of lipopolysaccharide, had increased survival when given a subsequent high-dose challenge of lipopolysaccharide. Thus, thioredoxin is associated with the development of endotoxin tolerance in mice. PMID- 11902262 TI - Overexpression of Bcl-2 in the intestinal epithelium improves survival in septic mice. AB - OBJECTIVES: The aim of this study was to determine whether decreasing intestinal epithelial apoptosis in sepsis would alter mortality rates. The roles of the antiapoptotic protein Bcl-2 and the "executioner" protease caspase-3 in sepsis induced gut cell death also were evaluated. DESIGN: Prospective, randomized, controlled trial. SETTING: Animal laboratory in an academic medical center. INTERVENTIONS: Transgenic mice that overexpress Bcl-2 throughout the small intestinal epithelium (n = 23) and littermate controls (n = 27) were subjected to cecal ligation and puncture (CLP) and followed for 8 days to assess survival. A second group of transgenic (n = 15) and littermate animals (n = 15) were subjected to CLP and were killed between 16 and 48 hrs postoperatively to assess for intestinal apoptosis and active caspase-3 staining. MEASUREMENTS AND MAIN RESULTS: Survival of transgenic animals was 83% 8 days after CLP compared with 44% for littermate controls (p < .005). Survival curves between the two groups of animals began diverging within 24 hrs. Overexpression of Bcl-2 was associated with a significant decrease in apoptosis between 16 and 24 hrs post-CLP (p < .05) as well as decreased staining for active caspase-3. CONCLUSIONS: Decreasing intestinal epithelial cell death via overexpression of Bcl-2 improves survival in septic mice. The gut may play a central role in the pathophysiology of sepsis. PMID- 11902263 TI - Inhibition of interleukin-6-activated janus kinases/signal transducers and activators of transcription but not mitogen-activated protein kinase signaling in liver of endotoxin-treated rats. AB - OBJECTIVES: Endotoxin-induced cytokines, such as interleukin-6, mediate systemic inflammatory responses through multiple cellular signaling pathways. Interleukin 6 is also responsible for the synthesis of acute phase proteins. Recent studies have shown that endotoxin can inhibit signal transducers and activators of transcription (STAT)-3 tyrosine phosphorylation in cultured cells, suggesting that this effect may limit the synthesis of acute phase proteins. The purpose of this study was to examine the effects of endotoxin on interleukin-6 activation of STATs and mitogen-activated protein (MAP) kinase pathways in rat liver in vivo. DESIGN: Controlled laboratory study. SETTING: Medical school laboratory. SUBJECTS: Specific pathogen-free male Sprague Dawley rats. INTERVENTIONS: Under anesthesia, interleukin-6 was injected into the portal vein of rats 4 hrs after the bolus intravenous administration of endotoxin (1 mg/kg) or saline. The effects of interleukin-6 on key intermediates in early steps of the interleukin-6 signaling pathway, including janus kinase-1, gp 130, the interleukin-6 receptor, STAT1, and STAT3, were examined in both saline and endotoxin-treated rats. MEASUREMENTS AND MAIN RESULTS: In endotoxin-treated rats, there was significant inhibition of interleukin-6 activation of janus kinase-1, gp 130, the interleukin 6 receptor, STAT1, and STAT3. These signaling changes were associated with decreased tissue abundance of interleukin-6 receptors and STAT3. In contrast to its effects on the janus kinase/STAT pathways, interleukin-6 activation of MAP kinases (extracellular signal-regulated kinase-1, extracellular signal-regulated kinase-2, and p38) was unaffected by endotoxin. CONCLUSIONS: The pathway-specific inhibition of interleukin-6 signaling responses in the liver may be an important determinant of the pathophysiologic consequences of endotoxin exposure. PMID- 11902264 TI - Effects of hypertonic saline, mannitol, and urea with regard to absorption and rebound filtration in cat skeletal muscle. AB - OBJECTIVE: To study the effects of the hypertonic solutions 15% mannitol, 3% and 7.5% saline, and 30% urea at clinically relevant plasma concentrations with regard to absorption and rebound effects on tissue volume in skeletal muscle. DESIGN: A prospective, experimental study. SETTING: University laboratory. SUBJECTS: Twenty-eight anesthetized cats. INTERVENTIONS: The study was performed on an autoperfused and denervated cat calf muscle placed in a fluid-filled plethysmograph. Muscle volume changes and capillary filtration coefficient (reflecting capillary fluid conductivity) were measured before, during, and after intra-arterial infusion (4 mL/hr) of the hypertonic solutions. Mannitol and 3% saline have the same osmolality and were compared specifically in an attempt to distinguish osmotic effects from those specific to the compound. MEASUREMENTS AND MAIN RESULTS: All solutions reduced muscle volume during the infusion (p < .05). The maximum volume reduction persisted after 2 hrs of infusion for 3% and 7.5% saline, whereas there was a tendency for volume recovery during the urea infusion and a complete recovery back to control for mannitol. After discontinuation of the infusions, the muscle volume increased for all four solutions, stabilizing at the initial control for 3% and 7.5% saline, whereas it increased to levels above control for mannitol and urea (p < .05). Capillary filtration coefficient was increased by hypertonic saline (p < .05) but was unaffected by mannitol and urea. CONCLUSIONS: The effectiveness of a hypertonic solution in reducing tissue volume and its tendency to cause a rebound volume increase depends not only on the osmolality of the solution. Hypertonic saline may in the long run be superior to mannitol and urea to increase plasma volume or decrease tissue volume of an organ, because it lacks rebound effects. Alterations in capillary filtration coefficient (fluid conductivity) may reflect volume changes of the capillary endothelial cell and thereby differences in cell membrane permeability for the hypertonic solutions, also consistent with the obtained differences in tissue volume effects. PMID- 11902265 TI - Antithrombin effects on endotoxin-induced microcirculatory disorders are mediated mainly by its interaction with microvascular endothelium. AB - OBJECTIVE: To investigate whether the protective effect of antithrombin III, which has been shown to exert beneficial effects during septic disorders, including reduction of endotoxin-associated leukocyte/endothelial cell interaction and capillary perfusion failure, is mainly based on its anticoagulant capacity or direct effects on the microvascular endothelium. DESIGN: Animal study with three treatment groups. SETTING: Animal research facility. SUBJECTS: Syrian golden hamsters, 6-8 wks old with a body weight of 60-80 g. INTERVENTIONS: In skinfold preparations of hamsters, normotensive endotoxemia was induced by intravenous administration of 2 mg/kg endotoxin (lipopolysaccharide, 2 mg/kg). Antithrombin III (n = 7 animals; 250 units/kg) or tryptophan49-blocked antithrombin III (n = 6; 250 units/kg) was substituted intravenously 5 mins before lipopolysaccharide administration. Saline-treated animals (n = 11), receiving only lipopolysaccharide, served as controls. Tryptophan49-blocked antithrombin III binds to glycosaminoglycans at the endothelial surface to a significantly lower extent while retaining its progressive anticoagulant effects. MEASUREMENTS AND MAIN RESULTS: Compared with controls, antithrombin III significantly reduced lipopolysaccharide-induced arteriolar and venular leukocyte adherence (p < .01) and prevented depression of functional capillary density (p < .01), whereas tryptophan49-blocked antithrombin III failed to significantly improve both variables. As measured in vivo by a monoclonal fluorescein isothiocyanate-labeled anti-antithrombin III antibody and intravital microscopy, the lack of effect of tryptophan49-blocked antithrombin III was associated with significantly lower antithrombin III/endothelium binding coefficients after 1 hr, 3 hrs, and 24 hrs of endotoxemia (p < .01). CONCLUSIONS: We conclude that specific antithrombin III interactions with cell-surface glycosaminoglycans on the endothelium rather than anticoagulant properties are the mechanism of antithrombin III-mediated attenuation of leukocyte/endothelial cell interaction and capillary perfusion failure. PMID- 11902267 TI - Endogenous lipoproteins impact the response to endotoxin in humans. AB - OBJECTIVE: To determine whether endogenous lipoproteins can abrogate the host response to lipopolysaccharide (LPS) in vivo. DESIGN: Randomized, placebo controlled study. SETTING: Urban public hospital with academic affiliation. SUBJECTS: Eighteen healthy, normolipidemic, normal weight volunteers, 21-35 yrs of age. INTERVENTIONS: Fasting and postprandial (hypertriglyceridemic) subjects were injected with endotoxin (LPS, Lot EC-5, 4 ng/kg = 20 endotoxin units/kg) as either a bolus or following preincubation of the LPS with autologous whole blood vs. saline. In addition, LPS-induced cytokine production was determined ex vivo to examine the capacity of fasting vs. hypertriglyceridemic whole blood to attenuate the effect of large, potentially lethal concentrations of LPS in humans. MEASUREMENTS: Vital signs were recorded and serial blood samples analyzed for changes in white blood cell count, cytokine, and stress hormone levels over 24 hrs. The distribution of lipoproteins in fasting and postprandial blood after preincubation was determined using 125I-LPS. MAIN RESULTS: Endogenous lipoproteins abrogated the host response to LPS in vivo, but only after preincubation with the LPS. Peak oral temperature (p < .05) and white blood cell count (p < .05), and plasma tumor necrosis factor (TNF)-alpha (p < .01) and adrenocorticotropic hormone levels (p < .03) were significantly reduced in volunteers injected with LPS preincubated with whole blood vs. LPS preincubated with saline. Approximately 80% of the LPS was bound to lipoproteins after preincubation with either fasting or hypertriglyceridemic blood. Thus, protection was associated with lipoprotein binding. In addition, hypertriglyceridemic but not fasting blood inhibited the ex vivo TNF-alpha response to large, highly toxic doses of LPS (p < .05). Without the preincubation of lipoproteins with LPS, there was a trend for an exaggerated clinical and TNF-alpha response in the hypertriglyceridemic subjects. CONCLUSION: Preincubation of LPS with whole blood promotes lipoprotein-LPS binding and is associated with an attenuated response to this toxic macromolecule. Although the clinical relevance of these data requires further elucidation, our results continue to support a lipid-based therapeutic strategy to combat gram-negative sepsis. PMID- 11902266 TI - Parental perspectives on end-of-life care in the pediatric intensive care unit. AB - OBJECTIVE: To identify priorities for quality end-of-life care from the parents' perspective. DESIGN: Anonymous, self-administered questionnaire. SETTING: Three pediatric intensive care units in Boston. PARTICIPANTS: Parents of children who had died after withdrawal of life support. MEASUREMENT AND MAIN RESULTS: Parents' views of the adequacy of pain management, decision making, and social support during and after the death of their child were measured with the Parental Perspectives Questionnaire. Of 96 eligible households, 56 (58%) responded. In 90% of cases, physicians initiated discussion of withdrawal of life support, although nearly half of parents had considered it independently. Among decision-making factors, parents rated the quality of life, likelihood of improvement, and perception of their child's pain as most important. Twenty percent of parents disagreed that their children were comfortable in their final days. Fifty-five percent of parents felt that they had little to no control during their child's final days, and nearly a quarter reported that, if able, they would have made decisions differently. There were significant differences (p < .001) between the involvement of family, friends, and staff members at the time of death and greater agreement (p < .01) about the decision to withdraw support between parents and staff members than with other family members. CONCLUSIONS: Parents place the highest priorities on quality of life, likelihood of improvement, and perception of their child's pain when considering withdrawal of life support. Parents make such decisions and garner psychosocial support in the context of a social network that changes over time and includes healthcare professionals, family, and friends. PMID- 11902268 TI - Randomized clinical trial of extended use of a hydrophobic condenser humidifier: 1 vs. 7 days. AB - OBJECTIVE: To determine whether extended use (7 days) would affect the efficiency on heat and water preservation of a hydrophobic condenser humidifier as well as the rate of ventilation-acquired pneumonia, compared with 1 day of use. DESIGN: Prospective, controlled, randomized, not blinded, clinical study. SETTING: Twelve bed intensive care unit of a university hospital. PATIENTS: One hundred and fifty five consecutive patients undergoing mechanical ventilation for > or = 48 hrs. INTERVENTIONS: After randomization, patients were allocated to one of the two following groups: a) heat and moisture exchangers (HMEs) changed every 24 hrs; b) HMEs changed only once a week. Devices in both groups could be changed at the discretion of the staff when signs of occlusion or increased resistance were identified. MEASUREMENTS AND MAIN RESULT: Efficient airway humidification and heating were assessed by clinical variables (numbers of tracheal suctionings and instillations required, peak and mean airway pressures). The frequency rates of bronchial colonization and ventilation-acquired pneumonia were evaluated by using clinical and microbiological criteria. Endotracheal tube occlusion, ventilatory support variables, duration of mechanical ventilation, length of intensive care, acquired multiorgan dysfunction, and mortality rates also were recorded. The two groups were similar at the time of randomization. Endotracheal tube occlusion never occurred. In the targeted population (patients ventilated for > or = 7 days), the frequency rate of ventilation-acquired pneumonia was 24% in the HME 1 day group and 17% in the HME 7-day group (p > .05, not significant). Ventilation acquired pneumonia rates per 1000 ventilatory support days were 16.4/1000 in the HME 1-day group and 12.4/1000 in the HME 7-day group (p > .05, not significant). No statistically significant differences were found between the two groups for duration of mechanical ventilation, intensive care unit length of stay, acquired organ system derangements, and mortality rate. There was indirect evidence of very little, if any, change in HME resistance. CONCLUSIONS: Changing the studied hydrophobic HME after 7 days did not affect efficiency, increase resistance, or altered bacterial colonization. The frequency rate of ventilation-acquired pneumonia was also unchanged. Use of HMEs for > 24 hrs and up to 7 days is safe. PMID- 11902269 TI - Adult long-segment tracheal stenosis attributable to complete tracheal rings masquerading as asthma. AB - OBJECTIVE: a) To report on an adult patient with congenital long-segment tracheal stenosis from complete tracheal rings complicated by tracheomalacia; b) to highlight the fact that some patients with airway narrowing could be misdiagnosed as having bronchial asthma; and c) to discuss our management with a custom-made tracheostomy tube extending to the carina. DESIGN: Case report. SETTING: A university hospital's 14-bed medical/surgical intensive care unit. PATIENT: A 21 yr-old patient, with a history of what was labeled as asthma, was admitted to the intensive care unit with diabetic ketoacidosis, pneumonia, respiratory failure, and septic shock. INTERVENTIONS: Her therapy included assisted mechanical ventilation through an endotracheal tube. Initially, a size 6.0 endotracheal tube was used. Finally, a custom-made tracheostomy tube extending to the carina was inserted to manage her persistent infantile trachea. MEASUREMENT AND MAIN RESULTS: During 4 months in the intensive care unit, she suffered numerous airway problems from her narrow trachea that were eventually attributed to congenital long-segment tracheal stenosis from complete tracheal rings. Bacterial pneumonia, viral tracheobronchitis, and tracheomalacia complicated her course. Multiple attempts at extubation failed and, after translaryngeal endotracheal tubes and tracheostomy tubes of decreasing size, her airway was managed with a size 5.0 custom-made tracheostomy tube with the tip extending to her carina. She was totally dependent on this tube. CONCLUSION: Airway narrowing may masquerade as asthma. Congenital tracheal stenosis is rare and is associated with a high mortality rate. Complete tracheal rings presenting in adulthood are extremely rare, and we report the first case of long-segment pantracheal stenosis presenting in adulthood. Surgical treatment with tracheoplasty is difficult. A custom-made tracheostomy tube to stent the entire trachea is one management option. Tracheal stenosis should be excluded in patients with a chronic lack of response to therapy for asthma. PMID- 11902271 TI - Cardiopulmonary resuscitation for pulseless rhythms and asystole. PMID- 11902270 TI - Late diagnosis of ornithine transcarbamylase defect in three related female patients: polymorphic presentations. AB - OBJECTIVE: To describe three female patients of one family with different phenotypes of the same mutation of the ornithine transcarbamylase gene. X-linked inherited ornithine transcarbamylase deficiency is the most frequent urea cycle disorder. Many of the hemizygous males die during the neonatal period. Women, who are mostly healthy carriers, can also develop symptomatic hyperammonemia. DESIGN: Case study. SETTING: Intensive care unit and internal medicine unit at a university hospital. PATIENTS: The 20-yr-old female propositus was hospitalized for unexplained coma. She had a history of headaches, recurrent vomiting, specific anorexia for high-protein foods, and an acute neurologic crisis with alleged food poisoning 8 yrs before. The present episode began with psychiatric symptoms and seizures treated by diazepam and valproate. This unexplained coma, associated with respiratory alkalosis and major brain swelling on brain computed tomography scan, revealed hyperammonemia leading to the diagnosis of ornithine transcarbamylase deficiency. Continuous venovenous hemodiafiltration and treatment with sodium benzoate and phenylbutyrate improved the situation. However, the patient had some neurologic sequelae. DNA studies have disclosed a pathogenic mutation in the ornithine transcarbamylase gene of the patient, her mother, and her sister. For the mother, the disease was overlooked despite the onset of unusual headaches and neurologic signs that mimicked a cerebral tumor 12 yrs before. The 28-yr-old sister of the propositus has always been asymptomatic, even during pregnancy. CONCLUSIONS: Diagnosis of urea cycle disorder should be considered in any patient with unexplained neurologic and psychiatric disorders with selective anorexia, even in adulthood. Unexplained coma with cerebral edema and respiratory alkalosis requires urgent measurement of ammonemia and metabolic work-up. PMID- 11902272 TI - Lisofylline: anti-acute respiratory distress syndrome or just anti-inflammatory? PMID- 11902273 TI - Benefiting from lipoprotein-endotoxin interactions. PMID- 11902274 TI - Source of inflammatory mediators in cardiopulmonary bypass. PMID- 11902275 TI - Stomach gas laboratory in critical care. PMID- 11902276 TI - A spoonful of sugar...? PMID- 11902277 TI - Elevation of plasma peptidoglycan and peripheral blood neutrophil activation during hemorrhagic shock: plasma peptidoglycan reflects bacterial translocation and may effect neutrophil activation. PMID- 11902278 TI - Modulation of types I and II acute phase reactants with insulin-like growth factor-1/binding protein-3 complex in severely burned children. PMID- 11902279 TI - Oxygen-induced acute hypercapnia in chronic obstructive pulmonary disease: what's the problem? PMID- 11902280 TI - Saline-induced hyperchloremic metabolic acidosis. PMID- 11902281 TI - Induction of apoptosis in sepsis: cell suicide may be beneficial. PMID- 11902282 TI - Cytokine signaling in sepsis: redundancy, crosstalk, and regulatory mechanisms. PMID- 11902283 TI - Starting the quest to define optimal pediatric end-of-life care. PMID- 11902284 TI - Clinical guidelines for the treatment of ventilator-associated pneumonia. PMID- 11902285 TI - Effect of the interleukin-6 promoter polymorphism (-174 G/C) on the incidence and outcome of sepsis. AB - OBJECTIVE: A biallelic polymorphism within the human interleukin (IL)-6 gene promoter region (-174 G/C) has been shown to affect IL-6 transcription in vitro and IL-6 plasma levels in healthy adults. Because IL-6 is excessively released into the circulation during sepsis and closely correlates with the clinical course, we studied whether this promoter polymorphism has an effect on the incidence and/or outcome of sepsis. DESIGN: Population-based association study in critically ill patients and healthy controls. SETTING: Surgical intensive care unit (ICU) in a German university hospital. PATIENTS: Surgical patients (n = 326) of German Caucasian origin with an ICU stay of at least 3 days admitted between 1997 and 1999 were prospectively enrolled. In a subset of 50 patients, sepsis was diagnosed according to consensus criteria (American College of Chest Physicians 1992). Healthy sex-matched adults of the same ethnic and geographic background served as controls. INTERVENTIONS: Blood sampling. MEASUREMENTS AND MAIN RESULTS: The (-174 G/C) polymorphism was genotyped by an allele-specific polymerase chain reaction. IL-6 plasma levels were determined by enzyme-linked immunosorbent assay. Genotype distribution and allele frequencies did not differ significantly between patients with or without sepsis and healthy controls. In patients who finally succumbed to sepsis, significantly less GG homozygotes were observed compared with survivors (p = .008). Median systemic IL-6 levels in septic patients closely correlated with outcome (p < .0001) but were not associated with the IL-6 promoter genotype. CONCLUSIONS: The IL-6 promoter polymorphism (-174 G/C) does not affect the incidence of sepsis. However, the GG homozygous genotype is significantly associated with an improved survival in sepsis. Because this association is independent from the systemic IL-6 response, we suggest that other genetically linked polymorphisms may be the primary cause. PMID- 11902286 TI - Continuous monitoring of gastric intraluminal carbon dioxide pressure, cardiac output, and end-tidal carbon dioxide pressure in the perioperative period in patients receiving cardiovascular surgery using cardiopulmonary bypass. AB - OBJECTIVE: To verify the hypothesis that the gastric intraluminal PCO2 (PgCO2) changes independently of the change in cardiac output (CO) during and after cardiovascular surgery using cardiopulmonary bypass (CPB), and that the elevation of PgCO2 affects the patients' morbidity. DESIGN: Prospective, noninterventional study. SETTING: Medical/surgical intensive care unit and operating theater of a university hospital. PATIENTS: Sixteen adults patients receiving elective cardiovascular surgery using CPB. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: After induction of anesthesia, the patients were fitted with a gastric tube equipped at the tip with a CO2 sensor (ion-selective field effect transistor) that can continuously measure real-time PgCO2, and a pulmonary artery catheter capable of monitoring continuous CO (CCO) and end-tidal CO2. Data from the devices was uploaded to a personal computer every 2 mins until the catheter was pulled off based on clinical judgment (PgCO2 values were blinded to everyone except the investigator). One patient expired as a result of multiple organ failure subsequent to sepsis, and postoperative morbidity assessed by the peak SOFA (sequential organ failure assessment) score (mean +/- SD 6.9 +/- 3.5; range, 2-13) was correlated with the peak PgCO2 during intensive care unit stay (mean +/ SD 74.1 +/- 30.7 mm Hg; range, 45-169 mm Hg) (p < .01, by regression analysis). The peak PgCO2 during surgery (mean +/- SD 71.1 +/- 18.1 mm Hg; range, 44-115 mm Hg) had no correlation with the postoperative morbidity. From analysis of CCO before, during, and after returning from the above 60 mm Hg of PgCO2, PgCO2 changed independently of CCO. CONCLUSIONS: PgCO2 changed independently of CCO, and its postoperative elevation was related to morbidity, even in the group of patients with a good outcome. Continuous monitoring of PgCO2 is useful for the detection of morbidity and can be expected to help elucidate the pathophysiology of change of PgCO2. PMID- 11902287 TI - Reliability of a new algorithm for continuous cardiac output determination by pulse-contour analysis during hemodynamic instability. PMID- 11902288 TI - Effect of an education program on decreasing catheter-related bloodstream infections in the surgical intensive care unit. AB - OBJECTIVE: The purpose of the study was to determine whether an education initiative aimed at improving central venous catheter insertion and care could decrease the rate of primary bloodstream infections. DESIGN: Pre- and postintervention observational study. SETTING: Eighteen-bed surgical/burn/trauma intensive care unit (ICU) in an urban teaching hospital. PATIENTS: A total of 4,283 patients were admitted to the ICU between January 1, 1998, and December 31, 2000. INTERVENTIONS: A program primarily directed toward registered nurses was developed by a multidisciplinary task force to highlight correct practice for central venous catheter insertion and maintenance. The program consisted of a 10 page self-study module on risk factors and practice modifications involved in catheter-related infections as well as a verbal in-service at staff meetings. Each participant was required to take a pretest before taking the study module and an identical test after its completion. Fact sheets and posters reinforcing the information in the study module were also posted throughout the ICU. MEASUREMENTS AND MAIN RESULTS: Seventy-four primary bloodstream infections occurred in 6874 catheter days (10.8 per 1000 catheter days) in the 18 months before the intervention. After the implementation of the education module, the number of primary bloodstream infections fell to 26 in 7044 catheter days (3.7 per 1000 catheter days), a decrease of 66% (p < .0001). The estimated cost savings secondary to the decreased infection rate for the 18 months after the intervention was between $185,000 and $2.808 million. CONCLUSIONS: A focused intervention primarily directed at the ICU nursing staff can lead to a dramatic decrease in the incidence of primary bloodstream infections. Educational programs may lead to a substantial decrease in cost, morbidity, and mortality attributable to central venous catheterization. PMID- 11902290 TI - Clinical utility of blood cultures drawn from central venous or arterial catheters in critically ill surgical patients. AB - OBJECTIVE: To determine the sensitivity, specificity, and predictive values of cultures done with blood drawn through a central venous or arterial catheter compared with peripheral venipuncture. DESIGN: Retrospective cohort study of critically ill surgical patients in whom samples for paired cultures were drawn through a central venous or arterial catheter and peripheral venipuncture. SETTING: Tertiary-care, university-affiliated medical center. PATIENTS: Two hundred seventy-one patients hospitalized on a surgical and a cardiothoracic intensive care unit between November 1994 and August 1997. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blinded assessments of culture results done by two physicians were used as the gold standard. Sensitivity, specificity, and positive and negative predictive values were compared for culture of blood from catheters and culture of blood from peripheral venipuncture. Of 499 observations, 426 were catheter-negative/venipuncture-negative, 19 were catheter-positive/venipuncture positive, 18 were catheter-negative/venipuncture-positive, and 36 were catheter positive/venipuncture-negative pairs. For catheter draws compared with peripheral venipuncture, sensitivity was 78% (confidence interval [CI], 65% to 90%) and 65% (CI, 50% to 79%) (p = .2), specificity was 95% (CI, 94% to 97%) and 98% (CI, 97% to 99%) (p = .002), positive predictive value was 63% (CI, 51% to 76%) and 78% (CI, 64% to 91%) (p = .1) and negative predictive value was 98% (CI, 96% to 99%) and 97% (CI, 95% to 98%) (p = .3). When central venous specimens as differentiated from arterial catheter specimens were compared with peripheral venipuncture, the difference between positive predictive values reached statistical significance (61% and 82%; p = .04). CONCLUSIONS: In critically ill surgical patients, cultures of blood drawn through a catheter are less specific than those obtained from a peripheral venipuncture. Both types of cultures have an excellent negative predictive value. Positive predictive value of cultures of blood drawn through a catheter is low and, when obtained from a central line, statistically less than from a peripheral venipuncture. Additional cultures seem to be necessary for the proper interpretation of a positive culture drawn through a catheter in critical care patients. PMID- 11902289 TI - Protein turnover, lipolysis, and endogenous hormonal secretion in critically ill children. AB - OBJECTIVES: The catabolic state is a major contributor to morbidity and mortality of critical illness and may be related to endocrine changes. We studied whether protein and lipid turnover correlate with insulin and growth and thyroid hormone plasma levels in critically ill infants. DESIGN: Prospective clinical study. SETTING: Pediatric intensive care unit. PATIENTS: Twelve critically ill children and ten age-matched controls. MEASUREMENTS: We measured lipolysis and protein turnover by infusing albumin-bound uniformly 13C palmitic acid and 2H3-leucine for 3 hrs and 2H5-glycerol for 5 hrs to critically ill infants. Simultaneously, we measured serum growth hormones, insulin, C-peptide, thyroid-stimulating hormone, T4, T3, albumin, retinol binding protein (RBP), and prealbumin. Hormone and serum protein levels were also measured in six children when recovered from critical illness. Ten healthy age-matched children served as controls for hormone serum levels comparison. RESULTS: Palmitic acid and glycerol turnover were 5.6 +/ 2.2 micromol/kg/min and 12.2 +/- 7.3 micromol/kg/min, respectively, whereas alpha-ketoisocaproic turnover was 4.9 +/- 2.8 micromol/kg/min. Alpha ketoisocaproic turnover positively correlated (R = 0.7, p = .03) with duration of pediatric intensive care unit admission and with prealbumin and RBP serum levels (R = 0.9, p = .001). Insulin-like growth factor binding protein (IGFBP)-2 was significantly higher and IGFBP-3 was significantly lower in critically ill children (p = .03 and p = .04 vs. recovery phase, respectively). No other hormonal differences were found. Serum albumin was significantly lower in sick children. We found a significant correlation between prealbumin and RBP and IGFBP 3 (R = 0.6, p = 0.03 and R = 0.6, p = .04, respectively). Alpha-ketoisocaproic turnover positively correlated with IGFBP-1 (R = 0.79, p = .01) and did not correlate with insulin-like growth factor I (R = -0.5, p = .15 [not significant]) No other correlations were found. Lipid turnover measurements did not correlate with any endogenous hormone levels or with duration of critical illness. CONCLUSION: Protein turnover but not lipolysis correlated with a persisting critically ill condition, serum prealbumin, RBP, and plasma IGFBP-1. PMID- 11902291 TI - Elevated expression of inducible nitric oxide synthase and inflammatory cytokines in the alveolar macrophages after esophagectomy. AB - OBJECTIVE: To evaluate the role of inducible nitric oxide (NO) synthase (iNOS) and inflammatory cytokines in alveolar macrophages (AMs) after esophagectomy in the pathogenesis of acute lung injury. DESIGN: Prospective, exploratory, open labeled clinical study. SETTING: Intensive care unit and operating room in a university hospital. PATIENTS: Thirteen patients receiving esophagectomy with carcinoma of the esophagus (postesophagectomy group), ten patients just before the surgery (preoperation group), and seven patents receiving surgery less invasive than esophagectomy (other surgery group) were selected. INTERVENTIONS: Bronchoalveolar lavage fluid (BALF) and blood samples were obtained from study groups. MEASUREMENTS AND MAIN RESULTS: The AMs in the BALF collected from each group were stained immunohistochemically with antibodies against iNOS, interleukin (IL)-6, and IL-8. The intensities of these expressions were determined by semiquantitative cytofluorometric system. NOx (NO2- + NO3-), IL-6, and IL-8 levels in the BALF and plasma were measured concurrently. The expressional intensities of iNOS, IL-6, and IL-8 in AMs obtained from the postesophagectomy group were maximal 24 hrs after the skin incision and significantly more evident than those from other groups. The IL-6, IL-8, and NOx levels in BALF and IL-6 and IL-8 levels in plasma in the postesophagectomy patients were also elevated. The intensities of iNOS and inflammatory cytokines expressions in AMs were closely related to postoperative respiratory failure. CONCLUSIONS: The activation of topical alveolar macrophages may be involved in the pathogenesis of pulmonary complications in the postoperative period after esophagectomy. PMID- 11902292 TI - Elevation of plasma peptidoglycan and peripheral blood neutrophil activation during hemorrhagic shock: plasma peptidoglycan reflects bacterial translocation and may affect neutrophil activation. AB - OBJECTIVE: To investigate the relations among bacterial transloation, plasma peptidoglycan elevation, and peripheral blood neutrophil activation during hemorrhagic shock. DESIGN: Prospective, randomized, unblinded animal study. SETTING: Surgical research laboratories of Shiga University of Medical Science. SUBJECTS: Male, specific pathogen-free Sprague-Dawley rats. INTERVENTIONS: The rats were randomly divided into three groups: a conventional group with normal intestinal flora (NF), an antibiotic (streptomycin and penicillin G) decontaminated group (AD), and a sham shock group with normal intestinal flora. The NF and AD groups were subjected to hemorrhagic shock (mean arterial pressure 30 mm Hg, for 30 to 90 mins). Rats were killed at 30, 60, and 90 mins after shock induction. Systemic blood and mesenteric lymph nodes (MLNs) were cultured for the determination of bacterial translocation (BT). Systemic plasma peptidoglycan and endotoxin concentrations were measured. To evaluate peripheral blood neutrophil activation, phagocytosis and hydrogen peroxide generation were assayed by flow cytometry. MEASUREMENTS AND MAIN RESULTS: In the NF group, BT to MLNs was significantly increased from 30 mins after shock induction. Blood culture and plasma endotoxin were positive at 90 mins but there were no significant differences. Assayed plasma peptidoglycan was significantly increased at 90 mins. Phagocytosis and hydrogen peroxide generation were significantly increased. Assayed plasma peptidoglycan concentrations showed significant positive correlations with the magnitude of BT to MLNs (r2 = .54) and hydrogen peroxide generation (r2 = .22) in individual animals. Furthermore, BT and these parameters were significantly suppressed in the AD group. CONCLUSIONS: First, we concluded that assayed plasma peptidoglycan reflects BT induced by hemorrhage because the increase in assayed plasma peptidoglycan was suppressed, as was BT, by antibiotic decontamination. Second, peripheral blood neutrophil activation was also suppressed when BT was prevented. We concluded BT to be involved in neutrophil activation. Our findings suggest hydrogen peroxide generation by neutrophils to be involved in plasma peptidoglycan elevation. PMID- 11902293 TI - Modulation of types I and II acute phase reactants with insulin-like growth factor-1/binding protein-3 complex in severely burned children. AB - OBJECTIVE: To determine whether 0.5 mg/kg insulin-like growth factor (IGF) 1/binding protein (IGFBP)-3, given intravenously, effectively alters the acute phase response in severely burned children. DESIGN: Longitudinal trial with each patient serving as their own control. SETTING: University-affiliated pediatric bum center. PATIENTS: Nine children, 15 yrs of age or less, with burns covering >40% of the total body surface area. INTERVENTIONS: Standard burn care with early burn wound excision and grafting. Blood sampled at defined time points before and after operative procedures. MEASUREMENTS AND RESULTS: Determination of types I and II acute phase reactant proteins, constitutive serum proteins, serum cytokines, serum IGF-1, IGFBP-3, and growth hormone levels. Treatment with IGF 1/BP-3 attenuated increases in type I (complement 3, alpha1-acidglycoprotein) and type II (haptoglobin, alpha1-antitrypsin) acute phase proteins. Further, IGF-1/BP 3 increased constitutive serum protein levels (prealbumin, retinol binding protein, transferrin) and decreased serum IL-6 levels. CONCLUSIONS: Low-dose IGF 1/BP-3 effectively attenuated the type I and type II hepatic acute phase response, increased serum levels of constitutive proteins, and modulated the hypermetabolic response. PMID- 11902295 TI - Regional variation in child mortality at hospitals lacking a pediatric intensive care unit. AB - OBJECTIVE: To investigate statewide variation in failure to utilize existing regional pediatric intensive care units (PICUs). METHODS: Deaths of children in hospitals lacking specialized units (non-PICU hospitals) were postulated to represent possible PICU utilization failures. A survey study was performed on hospital inpatient discharges and deaths in 1997, using data obtained from the New York Statewide Planning and Research Cooperative System (SPARCS). Children 0 14 yrs old were studied, excluding neonatal Diagnosis-Related Groups and emergency department deaths. Hospitals were considered to have a PICU if they had a board-certified pediatric intensivist on staff, and either New York State designation as a PICU or a separate dedicated unit for children. Non-PICU hospital pediatric death rates were compared for health service areas to determine whether regional variation occurred. RESULTS: Statewide, 157 of 584 (27%) pediatric hospital inpatient deaths occurred in non-PICU hospitals. Significant variation was seen among eight regions in pediatric death rates in non-PICU hospitals (p < .05). The 114 of 328 (35%) New York City inpatient deaths occurring in non-PICU hospitals significantly exceeded the 43 of 256 (17%) throughout the remainder of the state (p < .05). New York City non-PICU hospital death rates also were higher than in the rest of the state, when expressed per 100,000 pediatric population (8.04 vs. 2.00), and per 1,000 non-PICU hospital discharges (2.25 vs. 1.18), respectively (p < .05). Forty percent of New York City non-PICU hospitals experienced a pediatric inpatient death compared with only 13% in the rest of the state (p < .05). CONCLUSIONS: Although the death of some children in hospitals lacking a PICU is expected, the significant regional variation in these deaths suggests that local obstacles, perhaps unique to metropolitan areas, may interfere with access to existing pediatric critical care resources. PMID- 11902294 TI - Inhaled nitric oxide increases endothelin-1 levels: a potential cause of rebound pulmonary hypertension. AB - OBJECTIVE: Inhaled nitric oxide (iNO) is front-line therapy for pulmonary hypertension after repair of congenital heart disease. However, little clinical data exists regarding the effects of iNO on regulators of pulmonary vascular resistance. An imbalance between primary vasodilators, such as NO, and vasoconstrictors, such as endothelin-1 (ET-1), has been implicated in rebound pulmonary hypertension upon iNO withdrawal. The objective of this study was to determine whether iNO therapy alters plasma ET-1 levels. DESIGN: This is a prospective study involving pediatric and adult patients at risk for pulmonary hypertension. SETTING: Pediatric patients were in the cardiac intensive care unit and adult patients were in a tertiary-care hospital. PATIENTS: Group 1 included children with congenital heart disease requiring iNO for treatment of pulmonary hypertension after cardiopulmonary bypass (n = 15), group 2 was adults receiving iNO (n = 10), and group 3 included children at risk for pulmonary hypertension after bypass that did not require iNO (n = 8). INTERVENTIONS: Dosages of iNO were 2-60 ppm. The duration of therapy ranged from 23 to 188 hrs in group 1 and 29 to 108 hrs in group 2. MEASUREMENTS AND MAIN RESULTS: Arterial blood was obtained for the measurement of ET-1 levels before and during iNO therapy and 24 hrs after iNO withdrawal. Group 1 mean ET-1 levels increased to 127% of baseline by 12 hrs of iNO, remained elevated at 48 hrs (p < .05), then decreased to 71% of iNO levels 24 hrs after withdrawal (p < .01). Group 2 ET-1 levels increased to 147%, and 137% of baseline at 12 and 24 hrs of iNO therapy, then fell to 68% of baseline within 24 hrs of discontinuing iNO. ET-1 levels in group 3 decreased after surgery (p < .05). CONCLUSIONS: These data suggest that iNO increased plasma ET-1 levels, which subsequently decreased when iNO was discontinued. Increased circulating ET-1 levels might contribute to rebound pulmonary hypertension upon iNO withdrawal. PMID- 11902296 TI - Clinical trials and tribulations. PMID- 11902297 TI - Cancer clinical trials: risks and benefits. PMID- 11902298 TI - Fractures in pediatric Ewing sarcoma. AB - PURPOSE: To determine the incidence, timing, and clinical significance of long bone fractures in children with Ewing sarcoma family of tumors (ESFT). PATIENTS AND METHODS: We retrospectively reviewed 93 consecutive cases of ESFT of the long bones seen at a single institution over the course of a 37-year period. RESULTS: Fracture occurred in 14 (15%) of 93 patients with long-bone ESFT, most commonly in the femur. Approximately 30% of patients with tumors of the femur had fractures at some point in the course of their disease. The incidence of fracture was highest among patients with tumors of the proximal third of the femur (50%); these fractures were usually present at the time of initial diagnosis. Nine (64%) of the 14 fractures occurred after the start of radiotherapy, and three of these were associated with either local recurrence or second malignancy. CONCLUSIONS: Patients with femoral ESFT are at high-risk for fracture. If fractures occur after the completion of therapy, recurrence or second malignancy should be suspected. PMID- 11902299 TI - Visual improvement despite radiologically stable disease after treatment with carboplatin in children with progressive low-grade optic/thalamic gliomas. AB - BACKGROUND: The purpose of this study was to examine the clinical and radiologic response to carboplatin by children with progressive optic/thalamic gliomas. PATIENTS AND METHODS: Between July 1997 and July 1999, 12 consecutive children were treated with monthly carboplatin for progressive optic/thalamic gliomas. RESULTS: Five children have completed 12 cycles of carboplatin and five children are currently receiving treatment. Two children had progressive disease noted both clinically and radiologically. Nine children have stable radiologic disease and one child has had a partial radiologic response to chemotherapy. Eight children have had regular visual assessments. Four children (three with stable radiology and one with a partial radiologic response) have had improvement in their vision. Three children with radiologically stable disease have had no change in vision. One child has had deterioration in vision despite radiologically stable disease. CONCLUSIONS: The results suggest that the clinical response of optic/thalamic gliomas to carboplatin, as measured by visual acuity and visual fields, may be better than predicted by radiologic assessment. These data suggest that a prospective clinical study is warranted of the role of carboplatin in children with progressive optic/thalamic gliomas and visual impairment. PMID- 11902300 TI - Randomized, double-blind, crossover, placebo-controlled trial of intravenous ondansetron for the prevention of intrathecal chemotherapy-induced vomiting in children. AB - BACKGROUND: The purpose of this study was to determine the effectiveness of intravenous ondansetron in preventing vomiting after the administration of intrathecal chemotherapy in children. PATIENTS AND METHODS: Twenty-six children (ages 18 mo to 15 y) receiving intrathecal chemotherapy with either methotrexate or the combination of methotrexate, hydrocortisone, and Ara-C for the prophylactic treatment of central nervous system leukemia were randomly assigned to receive an infusion of normal saline or ondansetron at one of two doses (0.15 or 0.45 mg/kg) 30 minutes before undergoing the procedure. One hundred forty-six infusions were administered (51 placebo, 47 at the lower ondansetron dose, and 48 at the higher dose). Each patient acted as his or her own control, and each patient was studied at least three times. RESULTS: Twenty-three of 26 patients (88.5%) had postprocedural vomiting on at least one occasion. At least one episode of vomiting occurred during the 24 hours after the procedure in fifty-two of the procedures (35.6%). The incidence of vomiting was significantly greater after infusion of placebo than after either low-dose or high-dose ondansetron. The likelihood of severe vomiting was even more significantly reduced by the preadministration of ondansetron. Almost all of the intrathecal treatments associated with severe vomiting occurred after the infusion of placebo. CONCLUSIONS: Vomiting induced by intrathecal chemotherapy can be greatly reduced by the intravenous administration of ondansetron before the procedure, and severe vomiting can be virtually eliminated. PMID- 11902301 TI - Constitutional balanced chromosomal rearrangements and neoplasm in children. AB - A predisposition to tumor development is currently associated with some, but not all, constitutional chromosomal abnormalities. In a series of 578 children, in which conventional cytogenetic investigation was performed on material from various benign and malignant tumors, four boys and one girl were also found to have constitutional balanced chromosomal rearrangements. The figure of 5 in 578 is notable because the reported incidence of balanced rearrangements in newborns is approximately 1 in 450. Thereby suggesting that some, if not all, children with balanced constitutional chromosomal rearrangements have an increased predisposition for neoplasms developing. PMID- 11902302 TI - Comparison of multidimensional flow cytometry with standard morphology for evaluation of early marrow response in pediatric acute lymphoblastic leukemia. AB - PURPOSE: We compared multidimensional flow cytometry (MDF) with morphology in evaluating early marrow response to induction chemotherapy in pediatric ALL. METHODS: Chemotherapy response was determined by standard morphology or by MDF assessed by residual leukemic cell percentage remaining in the marrow on days 7, 14, and 28 of induction. Bone marrow response was classified as M3 (>25% leukemic blasts) or M1/M2 (< or = 25% leukemic blasts). Multidimensional flow cytometry evaluation was compared with that of standard morphology. Available day-7 and day 14 marrow slides were also reevaluated by a single pathologist without patients' clinical information. RESULTS: Of 46 day-7 specimens, eight (17%) had discordant MDF and morphologic results (P < 0.001), including six classified as M3 by morphology but were M1/M2 by MDF, and two were classified as M3 by MDF but were M1/M2 by morphology. Of 24 day-14 bone marrow specimens, five (20.5%) were discordant (P < 0.001), including two classified as M3 by morphology but were M1/M2 by MDF, and three were classified as M3 by MDF but were M1/M2 by morphology. Reevaluation of the blinded day-7 and day-14 marrow slides yielded discordance between repeated pathology readings of 11% (P < 0.001) and 6% (P = 0.04), respectively. CONCLUSION: Our data show significant discordance between the morphologic and MDF evaluation of early marrow response. Early response to therapy is a significant prognostic indicator in pediatric acute lymphoblastic leukemia and is used to alter subsequent treatment; thus, precise assessment of response is important. A larger comparison of MDF with morphology for the evaluation of early response, including correlation with clinical outcome, is warranted. PMID- 11902304 TI - Alpha-interferon therapy induces improvement of platelet counts in children with chronic idiopathic thrombocytopenic purpura. AB - PURPOSE: To investigate alpha-interferon (IFN) therapy for children with chronic idiopathic thrombocytopenic purpura (ITP). PATIENTS AND METHODS: Patients with refractory ITP lasting more than 12 months from diagnosis were included if they had platelet counts <50 x 10(9)/L and had received no treatment during the past month. Patients received IFN (3 x 10(6) U/m2 per dose), three times per week for 4 weeks; if partial (<150 x 10(9)/L) or no response was obtained, the same dose was continued for another 8 weeks. In patients with favorable response and subsequent decrease to pre-treatment values, an additional 4 weeks of treatment could be administered. RESULTS: Fourteen patients (ages 4-20 y) receiving 17 IFN courses were included. Mean initial platelet count was 29 +/- 15 x 10(9)/L. A significant increase was achieved during 14 of 17 courses (82.4%). All but two responses were transitory, and platelets returned to initial values after IFN discontinuation (mean 44 +/- 26 days). Considering the best response achieved by each patient, we observed: 1) 10 patients who achieved a sustained improvement of platelet count throughout the treatment period, decreasing to initial values after therapy was stopped; 2) one patient who achieved platelet count >150 x 10(9)/L, remaining with normal platelets at 18 months; 3) one patient who achieved platelet count >150 x 10(9)/L, remaining with platelets between 100 and 140 x 10(9)/L at 48 months; 4) one patient who had no response; and 5) one patient in whom therapy worsened the thrombocytopenia. A mild to moderate flu like syndrome and a moderate decrease of the absolute neutrophil count were the only side effects observed. CONCLUSION: Interferon therapy induces a significant increase of platelet count and seems to be a valid alternative therapy to attempt the achievement of prolonged remission in refractory ITP, to defer splenectomy in younger children, or to improve platelet count before planned splenectomy. PMID- 11902303 TI - Low relapse rate in children with acute lymphoblastic leukemia after risk directed therapy. AB - PURPOSE: Even though acute lymphoblastic leukemia (ALL) responds well to chemotherapy, relapse remains the major problem. This study documents relapse and survival rates in 85 consecutive children (33 at good risk, 52 at high risk) with ALL diagnosed in 1991 to 1996. PATIENTS AND METHODS: Until 1993, the New York II protocol for the high-risk group and a combination of UKALL XI (induction) and R blocks of ALL-REZ BFM-87 (intensification) regimens for patients at good risk were used. To reduce toxicity, the protocols were subsequently modified. Consolidation treatment was the same for both groups, consisting of a lower cytarabine dose and methotrexate removal, whereas intensification was changed only for the high-risk group using the BB block of the NHL-BFM-90 protocol. The bone marrow clearance of leukemia was assessed on day 22, and minimal residual disease was detected using polymerase chain reaction analysis of Ig heavy-chain gene rearrangements. RESULTS: Seventy patients had common precursor B lineage ALL, six had pre-B-ALL, eight had T-ALL, and one had B-ALL. Two patients never achieved remission and died. Six patients died of consolidation-related complications. Four more patients died, two during induction and two during maintenance therapy. Two other children had relapse (2.3%), both of whom were treated with the earlier protocols and then underwent bone marrow transplantation. Four more children with morphologically complete remission showed minimal residual disease (which reached the levels of 1 leukemic cell among 10(2)-10(4) normal cells) with the use of clone-specific probes at several points of the study intervals, but never had relapse. The 5-year overall and event-free survival rates were 86% and 83%, respectively. The 5-year overall survival rates for good-risk and high-risk groups were 94% and 81%; the corresponding event-free rates were 91% and 78%. The 5-year event-free survival rate in the patients at high risk was significantly higher after the protocol change (90% vs. 65%, P = 0.04). CONCLUSIONS: The modification proved to be effective in diminishing the therapeutic toxicity and improving the efficacy, mainly for the high-risk group. PMID- 11902305 TI - Effect of hydroxyurea therapy on resting energy expenditure in children with sickle cell disease. AB - Effects of hydroxyurea therapy on resting energy expenditure (REE) in children with sickle cell disease have not been evaluated. Eight children with sickle cell disease were examined before hydroxyurea therapy and again 6.9 +/- 3.5 months after hydroxyurea initiation. Resting energy expenditure, dietary intake, and growth were assessed. In six children, baseline REE was elevated, and REE decreased an average of 17% with hydroxyurea. This was associated with a significant increase in fetal hemoglobin. These pilot data suggest that hydroxyurea may curtail the hypermetabolic state observed in children with sickle cell disease and may offer a clinically important secondary benefit. PMID- 11902306 TI - Renal cell carcinoma as a secondary malignancy after treatment of acute promyelocytic leukemia. AB - Numerous children have been treated successfully for cancer and are surviving into adulthood. As this population has aged, an increasing number of secondary malignancies has emerged. Renal cell carcinoma (RCC) is a rare tumor in childhood and has not been documented previously to occur after treatment of acute promyelocytic leukemia (APL). This report describes the clinical course of APL treated in a child in whom RCC subsequently developed during adolescence approximately 5 years after therapy. PMID- 11902307 TI - Stage 4 squamous cell carcinoma of the tongue in a child: complete response to chemoradiotherapy. AB - This report describes a complete response to a chemoradiotherapy regimen in a child with an advanced and unresectable squamous cell carcinoma of the tongue. An 8-year-old girl had stage 4 squamous cell carcinoma of the tongue (T4N2M0), causing severe trismus and dysphagia. She received hyperfractionated external beam radiotherapy (total 74.4 Gy) and concomitant intravenous infusion of hydroxyurea (0.313 mg/m2 per min) for 43 days. Grade 3 mucositis and myelosuppression were the main toxicities. There was marked symptomatic improvement, and the patient achieved a complete response. She is disease-free 24 months after treatment, and all the acute symptoms have resolved. The regimen was well tolerated with acceptable toxicity and led to a complete objective response. This regimen needs further evaluation to confirm its efficacy and to ascertain its long-term effects in children. PMID- 11902308 TI - Long-term treatment with ketamine in a 12-year-old girl with severe neuropathic pain caused by a cervical spinal tumor. AB - A 12-year-old girl presented with head and neck pain, myoclonic movements, and decreased strength in all extremities caused by a cervical spinal tumor (glioblastoma multiforme). A partial resection of the tumor was performed. Three weeks later, she had superficial pain distributed in all dermatomes below her cervical medullary lesion. Touch (e.g., gentle hugs from relatives) and movements elicited paroxysm of intense pain. The pain was not relieved by increased doses of morphine. A test dose of ketamine (7.5 mg intravenous) provided an abrupt decrease in pain intensity, and continuous infusions of subcutaneous morphine and intravenous ketamine were started. Benzodiazepines were administered to avoid psychotomimetic effects from ketamine and to diminish myoclonic movements. The doses of analgesics and benzodiazepines were increasingly titrated (subcutaneous morphine 163-750 mg/24 hr, intravenous ketamine 36-410 mg/24 hr, subcutaneous midazolam 5-20 mg/24 hr, and intravenous diazepam 11.5-122.5 mg/24 hr) until her death 67 days after start of ketamine. She remained awake until the last day before her death. For the last 29 days of life, the pain treatment regimen was successfully continued in her home (400-km distance from the hospital). In conclusion, this case demonstrates that ketamine treatment may be effective in children with severe neuropathic pain not responsive to other analgesics. This patient also demonstrates the feasibility of long-term ketamine treatment in pediatric oncology and that such treatment can be administered in a home care setting. PMID- 11902309 TI - Complex regional pain syndrome in pediatric patients with severe factor VIII deficiency. AB - Two boys with severe factor VIII deficiency that initially presented with acute onset of joint pain and swelling consistent with an uncomplicated hemarthrosis are reported. When appropriate management failed to provide resolution of symptoms, alternate diagnoses were considered. Both boys ultimately had complex regional pain syndrome (CRPS) diagnosed. The delay in diagnosis contributed to prolonged patient discomfort and lack of appropriate therapy. Complex regional pain syndrome encompasses a group of disorders that are characterized by pain severity or duration disproportionate to that expected. It is uncommon in the pediatric population. Because early diagnosis and appropriate treatment may improve outcome, it is important for practitioners to consider CRPS in the differential diagnosis of persistent pain in children with hemophilia. PMID- 11902310 TI - Hemophagocytosis and granulomas in the bone marrow of a child with Down syndrome. AB - Persistent fever with pancytopenia and hepatomegaly with negative blood cultures and no obvious focus of infection in a child with Down syndrome should arouse a suspicion of leukemia. Bone marrow examination and clot biopsy from one such patient revealed hemophagocytosis and granulomas, with serologic evidence of recent Epstein-Barr virus infection. Bone marrow granulomas are not a feature of Epstein-Barr infection. Later, bone marrow culture and repeat blood culture grew Salmonella typhi. Thus, in a febrile child, when performing a bone marrow aspirate, a clot biopsy and culture for infectious etiology may be helpful, even when leukemia is strongly suspected. PMID- 11902311 TI - Herpesvirus-6 encephalitis complicated by Wernicke-Korsakoff syndrome in a pediatric recipient of unrelated cord blood transplantation. AB - A 10-year-old girl with M2 acute myeloid leukemia underwent an unrelated cord blood transplantation in refractory first relapse. On day +13, after 48 hours with fever, she showed a measles-like rash, and on day +15, she began experiencing neurologic symptoms (headache, tremors, weakness, nystagmus, mild confusion, speaking, taste, and behavior disturbances, and focal seizures). She also had amnesia for recent events with disability to learn, mimicking Wernicke Korsakoff syndrome. Computed tomography of the brain and cerebrospinal fluid (CSF) and electroencephalogram were nonspecific. We found human herpesvirus 6 (HHV-6) DNA in CSF and cytomegalovirus in bronchoalveolar lavage using polymerase chain reaction techniques. Treatment with ganciclovir and foscarnet was effective, with total resolution of symptoms. PMID- 11902312 TI - Successful treatment of refractory Langerhans cell histiocytosis with unrelated cord blood transplantation. AB - A 2-month-old girl presented for treatment with a diffuse rash, interstitial pneumonia, otorrhea, and lymphadenopathy. Skin biopsy confirmed Langerhans cell histocytosis by electron microscopy. After receiving multiple courses of chemotherapy, only marginal improvement was achieved, with progressive marrow and liver involvement. The decision was made to pursue a human leukocyte antigen identical unrelated cord blood transplantation. Two years after transplant, the bone marrow was clear of Langerhans cell histocytosis and 100% donor engraftment. The poor prognosis of patients with an inadequate response to therapy and the presence of organ dysfunction (marrow and liver) substantiated the decision to pursue an unrelated cord blood transplantation. PMID- 11902313 TI - Unrelated cord blood transplantation for an infant with chemotherapy-resistant progressive Langerhans cell histiocytosis. AB - The authors describe a patient successfully treated with unrelated cord blood transplantation (CBT) for chemotherapy-resistant progressive Langerhans cell histiocytosis (LCH). An 8-month-old boy had LCH diagnosed based on the histologic examination of skin lesions. Despite intensive chemotherapy and immunotherapy, the disease was progressive, with organ dysfunction. He received unrelated CBT after a conditioning regimen consisting of total body irradiation, etoposide, and melphalan. He was in complete remission 12 months after the transplantation. The authors suggest that CBT could be considered in the treatment of patients with chemotherapy-resistant progressive LCH, especially if there are no available human leukocyte antigen-matched family donors. PMID- 11902314 TI - Psychiatric disorders before and after surgery for epilepsy. AB - PURPOSE: To examine the frequency of psychiatric disorders before and after surgery for epilepsy and the postoperative changes. METHODS: We examined the psychiatric status of 226 patients aged 18 years or older and with IQ >50 who underwent focal resection surgery after age 15 years at our center and were followed up for >2 years after surgery. RESULTS: Seventy-eight (34.5%) patients had psychiatric disorders before and/or after surgery, comprising 36 patients with psychosis, 15 with affective disorders, and 27 with neurotic/behavioral disorders. All but four of these patients had temporal lobe epilepsy. Of the 61 patients who had psychiatric disorders before surgery, 22 patients were free of psychiatric symptoms after surgery (eight with psychosis, 13 with neurotic/behavior disorders, one with affective disorders), and 39 patients continued having psychiatric symptoms after surgery (22 with psychosis, 12 with neurotic/behavior disorders, five with affective disorders). Nine patients had a transient affective disorder that appeared 1-2 months after surgery and disappeared within 1-2 months. In eight patients, chronic psychiatric symptoms manifested after surgery (psychosis in six cases, neurotic/behavior disorders in two), one of whom exhibited postictal psychosis after drug noncompliance. Psychosis that disappeared after surgery was often seizure related. The patients with aggravated psychiatric conditions after surgery had a lower age at epilepsy onset and at surgery, and exhibited deviated personality traits even before surgery. CONCLUSIONS: One third of the surgical patients had psychiatric disorders before and/or after surgery. Preexisting psychiatric problems disappeared after surgery in one third of the patients, but persisted after surgery in the remaining patients, with aggravation in a few. Transient affective disorders appeared in some patients immediately after surgery, and chronic psychiatric disorders appeared after surgery in a minority of patients. The patients having aggravated psychiatric conditions after surgery tended to be young and to have deviated personality traits. The social situation of the patients having psychiatric disorders after surgery was not good. PMID- 11902315 TI - Psychiatric and neuropsychological problems in epilepsy surgery: analysis of 100 cases that underwent surgery. AB - PURPOSE: For the past 20 years (1978-1997), a series of 100 cases of uncontrolled epilepsy had surgery in our department under the stated standard for surgical indications and were followed up for 2-22 years after surgery. METHODS: We evaluated 70 cases of temporal lobectomy, 20 cases of neocortical focal resection, and 10 cases of corpus callosotomy. RESULTS: Analysis of postoperative seizure control showed that 78 cases were class 1 or 2 (no or rare seizures), 14 cases were class 3 (worthwhile improvement), and eight cases were class 4 (no improvement). As generally accepted, temporal lobectomy was the most effective operative procedure, yielding excellent or good results in 87% of the 70 cases so treated. Among the nine cases in whom various psychiatric symptoms developed after surgery, four cases showed neurotic and five cases psychotic symptoms. Patients with psychosis had delusions of various types as a core symptom, combined with other symptoms such as anxiety, irritability, aggression, and depressive state. In two patients with psychosis who had episodes of delusions in the interictal phase before surgery, the symptoms were extremely resistant. The full IQ score of the Wechsler Adult Intelligence Scale-Revised (WAIS-R) was increased after temporal lobectomy in 75% of the cases (p < 0.01; n = 44). The general MQ score in 31 cases, however, showed a 50:50 split between increase and decrease postoperatively. In correlation with the dominance of language by the Wada test, the general MQ score in the 15 cases of nondominant temporal lobe resection showed a significant increase (p < 0.05); whereas the MQ in the 16 cases of dominant-side operation did not change. CONCLUSIONS: Our new test, in which hippocampal stimulation and supraspan learning are combined, seems to be efficient for estimating the postoperative outcome of memory function. PMID- 11902316 TI - Predisposed susceptibility and partial seizure disorder. PMID- 11902317 TI - Age-related clinical and neurophysiologic characteristics of intractable epilepsy associated with cortical malformation. AB - PURPOSE: To elucidate the relationship between the characteristics of cortical malformation (CM) and those of associated epilepsy, and also to investigate the prognostic value of the clinical and magnetic resonance imaging (MRI) findings for the seizure and mental outcome. METHODS: We studied 41 patients with CM and epilepsy, and the patients were divided according to the age at onset of epilepsy into two groups: one group of 15 patients with very early onset before age 3 months, and the other group of 26 patients with onset at 3 months or later. Statistical relationship was examined between the types of dysplastic lesions demonstrated by MRI and the age at onset of epilepsy. The effects of the onset age and the features of CM on the outcome of seizures and mental or developmental state also were analyzed. RESULTS: The very early-onset epilepsy was related to the dysplastic patterns of reduced sulci and blurred cortical-subcortical junction, which suggested focal CM, whereas the later-onset epilepsy was related to polymicrogyria. The age at onset of epilepsy was related to the poor seizure outcome, and both the onset age and wide distribution of CM were related to severe retardation. CONCLUSIONS: The type of CM influences the expression of associated epilepsy, especially its age-related features. The age at onset of epilepsy plays an important role in the seizure and mental outcome. PMID- 11902318 TI - Neuroradiologic findings in focal cortical dysplasia: histologic correlation with surgically resected specimens. AB - PURPOSE: We investigated the neuroradiologic characteristics of focal findings of surgically resected specimens obtained from 47 patients with focal cortical dysplasia (FCD). METHODS: Forty cases were detected by magnetic resonance imaging (MRI), and two cases were detected only by single-photon emission computed tomography (SPECT), but five cases could not be detected before operation. RESULTS: MRI revealed abnormal gyri and sulci in 34 patients (pachygyric in 18, polymicrogyric in 10, both in six), and blurring of the gray matter-white matter junction in 29 (72%) patients. Signal abnormalities were found in 36 (90%) patients, in the gray matter in 32, with white matter in 30, and at the gray matter-white matter junction in 13. Moreover, peculiar patterns of abnormal signals in the white matter were recognized, including remarkably abnormal subcortical signals of T2 hyperintensity and T1 hypointensity adjacent to the dysplastic cortex in 15 cases, high radiated T2 signals extending from the ependymal surface of the lateral ventricle to the overlying cortex in 11 cases, and widespread abnormal signals in the white matter with gray matter involvement in four cases. Histologically, these abnormal signals corresponded to various degrees of dyslamination and morphologic abnormalities of neurons and glial cells in the gray matter, and to dysmyelination, ectopic clustering of dysplastic neurons, glial proliferation, and necrotic change in the white matter. Regional cerebral blood flow SPECT showed interictal hypoperfusion in 29 (62%) of the 47 patients, interictal hyperperfusion in two, and ictal hyperperfusion in 28 of the 34 patients associated with FCD. [123I]iomazenil SPECT demonstrating the distribution of central benzodiazepine receptors showed low accumulations localized spatially corresponding to the epileptogenic foci associated with FCD in seven of eight patients. CONCLUSIONS: These results demonstrate that neuroimaging reflects various structural and functional changes closely related to epileptogenesis in FCD. PMID- 11902319 TI - Intractable epilepsy: definition and neurobiology. PMID- 11902320 TI - Surgical strategy and outcomes for epileptic patients with focal cortical dysplasia or dysembryoplastic neuroepithelial tumor. AB - PURPOSE: The purpose of this study was to clarify and compare the influence of surgical strategy on relief from seizures in patients with focal cortical dysplasia (FCD) and those with dysembryoplastic neuroepithelial tumor (DNT). METHODS: Six patients with FCD and five patients with DNT, all of whom underwent surgical resection for medically intractable epilepsy, were compared in terms of presurgical seizure types and frequency, location of lesions, magnetic resonance imaging (MRI), single-photon emission computed tomography (SPECT) with 99mTc-ECD, scalp electroencephalogram (EEG), and long-term video-EEG recording. Prolonged subdural recordings and intraoperative electrocorticograms (ECoG) were analyzed. The influences of surgical strategies on seizure outcomes were retrospectively analyzed. RESULTS: In all the FCD patients, ictal SPECT revealed hyperperfusion in the regions where MRI showed FCD. Interictal epileptiform activity and ictal seizure onset on ECoG performed with subdural electrodes were localized on the FCD itself. In contrast, the tumors of all the DNT patients were depicted as hypoperfuse areas on interictal SPECT scans. Ictal SPECT in one DNT patient showed hyperperfusion in the area enclosing the tumor. Interictal spiking in all DNT patients and ictal seizure onset in two DNT patients were not in the lesions themselves but in an area enclosing the lesion. All but one patient with FCD who underwent total lesionectomy became seizure free. All DNT patients who underwent resection of the epileptogenic cortex associated with lesionectomy became seizure free or achieved a 90% reduction in seizures. CONCLUSIONS: FCD has intrinsic epileptogenicity, whereas DNT is encompassed by epileptogenic cortical areas. Therefore, total lesionectomy is an essential strategy for FCD, whereas resection of the epileptic focus associated with lesionectomy of a DNT lesion is necessary to control seizures. PMID- 11902322 TI - Neuropsychology of epilepsy. AB - PURPOSE: The neuropsychological approach to epilepsy is indispensable for assessment of cognitive function in an interictal period including pre- and postsurgical evaluation, and for disclosing the semiology of nonconvulsive status epilepticus. Another use of the neuropsychological approach is to identify a seizure-precipitating factor by loading systematic cognitive tasking, termed "neuropsychological EEG activation" (NPA), during standard EEG recordings. METHODS: In this study, NPA tasks consisted of reading, speaking, writing, written arithmetic calculation, mental arithmetic calculation, and spatial construction. RESULTS: The NPA tasks provoked epileptic discharges in 7.9% of the 480 epileptic patients and were often accompanied by myoclonic seizures. Among the cognitive tasks, mental activities mainly associated with use of the hands [i.e., writing (68.4%), written calculation (55.3%), and spatial construction (63.2%)] provoked the most discharges. Seizure-precipitating mental activities were found to be almost exclusively related to idiopathic generalized epilepsies (IGEs). CONCLUSIONS: These results suggest that NPA is a useful tool for examining the relationship between cognitive function and epileptic seizures, and that the IGE patients with myoclonic seizures are vulnerable to higher mental activity. PMID- 11902321 TI - Neuropsychological changes after surgical treatment for temporal lobe epilepsy. AB - PURPOSE: The effect of unilateral temporal lobectomy on seizure frequency is well recognized, but little is known about the neuropsychological changes that occur after surgical treatment. We assessed neuropsychological status in 26 patients with an average age of 35 years before and after unilateral temporal lobectomy for medically intractable TLE. METHODS: Neuropsychological examination to assess cognitive function, memory, attention, visuospatial analysis, language, and emotional functions was performed preoperatively and at 1 month and 1 year after the surgery. RESULTS: At both 1 month and 1 year after the surgery, the patients had improved scores, compared with the preoperative scores, on the Wechsler Adult Intelligence Scale-Revised (WAIS-R: verbal IQ, performance IQ, and full-scale IQ), Wechsler Memory Scale-Revised (WMS-R: verbal, general, and delayed paired associates memory), and Raven Colored Progressive Matrices. In the Minnesota Multiphasic Personality Inventory (MMPI), significant decreases were observed at 1 year after the surgery in the scores for infrequency, hypochondriasis, psychasthenia, and schizophrenia. Patients in whom the seizures had been relieved postoperatively also had improved scores on the WAIS-R, WMS-R, and Raven Colored Progressive Matrices. CONCLUSIONS: These data suggest that neuropsychological improvement postoperatively is influenced by the reduction in the frequency of seizures after surgery. PMID- 11902323 TI - Developmental assessment-based surgical intervention for intractable epilepsies in infants and young children. AB - PURPOSE: To define the most appropriate time for surgery for medically intractable epilepsies in infants and young children. METHODS: First we examined retrospectively the changes in developmental quotients (DQs) during the clinical course and the clinical factors affecting the DQ in 39 consecutive patients younger than 15 years, who underwent surgical treatment for intractable epilepsy. Second, we examined prospectively five new patients for early detection of developmental arrest or regression by periodic developmental assessments and whether this could lead to early surgical intervention, eventually resulting in minimal developmental defects. RESULTS: Retrospective studies revealed that the DQ progressively decreased with age and that the reduction of DQ was related to continuing frequent seizures in many patients. The prospective studies demonstrated that periodic developmental assessments could detect the reduction of DQ at 5 months or later after onset of frequent seizures in three patients. In two other patients, operations were performed before reduction of DQs, and their postoperative DQ levels were normal. The post-operative recovery of DQ was complete in one patient whose operation was performed 3 months after reduction of DQ, whereas it was incomplete in two others whose operations were carried out at 12 and 14 months after reduction, respectively. Furthermore, three patients with normal developmental outcome had shorter periods between the onset of frequent seizures and the operation (< or = 7 months) than those of two patients with developmental delay (> or = 17 months). CONCLUSIONS: To minimize the developmental defects, periodic developmental assessments should be initiated when frequent seizures have occurred, and surgery should be considered as soon as possible when DQ reduction is recognized. PMID- 11902324 TI - Special needs of the adolescent with epilepsy. AB - Adjustment to the inherent changes of adolescence is a challenge for many children, even healthy ones. Epilepsy may make successful adjustment even more difficult by disrupting self-control, adversely affecting the ability to conform to one's peers, and interfering with independence. Epilepsy and its treatment may affect education, employment, use of alcohol and illicit drugs, sexual activity, and pregnancy. These issues, in turn, may affect seizure control. Matching the medicine to the patient is particularly important in teenagers, a group of patients with special needs. Common seizure syndromes presenting in adolescence are briefly reviewed and used to illustrate key issues that may arise in the selection of an antiepileptic medication (AED). Compliance, interaction with other medications including oral contraceptive medications, cosmetic effects, teratogenicity, and impact on behavior are all factors to be considered. Newer medications may have some advantages over older compounds in this regard, and evidence of their utility is reviewed in the specific context of the epilepsy syndromes. In addition to prescribing medication, other important parts of the therapeutic management involve education, counseling, support, and advocacy. To identify issues and to provide appropriate counseling, it is beneficial to devote some of the office visit to an independent interaction with the patient, apart from the parents. One study questionnaire indicated that nurse specialists may be particularly valuable in supporting and meeting the needs of patients in this age group. Some centers have developed elegant methods of helping the patient make the transition from child neurology to adult neurology. PMID- 11902325 TI - Elderly patients with systemic disease. AB - Although the healthy elderly seizure patient can often be treated with antiepileptic drugs (AEDs) similarly to a younger counterpart, the elderly with systemic disease require a more complex approach. The interplay of multiple organ dysfunctions and multiple non-AED medications greatly increases the challenges of managing the majority of older epilepsy patients. In addition, age-related physiologic changes in hepatic and renal function, volume of distribution, and exaggerated sensitivity to side effects alter the usual "rules" for administering AEDs. Thorough knowledge of the general principles of geriatric physiology and pharmacology may predict optimal selection of initial or subsequent AED therapy in this population. PMID- 11902326 TI - Cognitive and behavioral effects of epilepsy treatment. PMID- 11902327 TI - The challenge of treatment selection for epilepsy. PMID- 11902328 TI - Treatment of benign epilepsy syndromes throughout life. AB - Classification of epilepsy has been refined to a degree that begins to allow identification of syndromes that are benign, if not in clinical pattern, at least with regard to ultimate outcome. Infants, children, adolescents, and even adults, on occasion, develop seizures that challenge judgment regarding selection of drugs and duration of treatment. Childhood absence epilepsy tends to follow a benign course, with abatement of need for treatment in many patients. However, in approximately 40% of patients with onset of this syndrome, there is evolution to juvenile myoclonic epilepsy (JME), a form of seizure disorder that requires life long treatment. One syndrome that is clearly almost always benign is benign epilepsy with centrotemporal spikes (BECTS). This clinical problem is defined by complaints, observations of behavior, and by a specific electroencephalographic pattern. Some patients are not treated; many undergo brief administration of medication. Infantile convulsive seizures that may be partial or with myoclonus tend to be familial and require treatment for 1 or 2 years. Time is required to make a decision about treatment because observation over time is required to be sure patients are developing normally, a hallmark of these syndromes. Familial groupings of partial seizures in adolescents appear to follow a benign course, as do those in many elderly patients with seizures after cerebral infarction. Treatment decisions require clinical judgment and observations over time. Drug selection is governed more by experience of individual clinicians, because benign syndromes occur infrequently and prospective studies are seldom performed. PMID- 11902329 TI - Importance of CpG dinucleotides in activation of natural IFN-alpha-producing cells by a lupus-related oligodeoxynucleotide. AB - The oligodeoxyribonucleotide (ODN) 5'-TTTTCAATTCGAAGATGAAT-3' (ODN H), identified in systemic lupus erythematosus (SLE) serum, induced the production of interferon (IFN)-alpha in human peripheral blood mononuclear cells (PBMC) when combined with lipofectin. Flow cytometric analysis with staining for surface antigens and intracellular IFN-alpha, showed that the IFN-alpha-producing cells (IPC) were the natural IPC, also termed type 2 dendritic cell precursors (pDC2) or plasmacytoid monocytes. The importance of unmethylated CpG dinucleotides for the interferogenic activity of ODN was studied. Methylation of CpG impaired the activity of single-stranded (ss) ODN H, but increased that of the complementary ssODN I. Furthermore, CpG-methylated double-stranded (ds) ODN Hmet-Imet lost, but hemimethylated dsODN H-Imet retained interferogenic activity. Inversion of the CpG to GpC had no effect on the interferogenic activity of ssODN H, increased that of ssODN I, however abolished the activity of dsODN H-I. Alteration of the CpG in ODN H to ApG and in the ODN I to CpT destroyed their activity. The induction of IFN-alpha is therefore sequence-specific, but unmethylated CpGs are not always required, especially not in ssODNs. Interferogenic DNA sequences could therefore be more frequent in eukaryotic genomes than previously thought and their capacity to activate natural IPC may have implications for immune responses to microbial antigens and nuclear autoantigens. PMID- 11902330 TI - The role of surface ig binding in the activation of human B cells by phosphorothioate oligodeoxynucleotides. AB - Phosphorothioate oligodeoxynucleotides (sODNs) can induce T-cell-independent polyclonal activation of human B cells by a mechanism that depends on both sequence and back-bone structure. Because matrix-bound as well as soluble sODNs are mitogenic, this stimulation may result from the engagement of surface receptor(s). In order to investigate whether surface immunoglobin (Ig) could be a receptor for sODNs, the interaction of sODNs-fluorescein isothiocyanate (FITC) with Ig-coated beads was examined. sODNs specifically bound to human IgM and IgG. Moreover, binding of sODN to human B cells induced temperature-dependent capping of bound receptors and colocalization of FITC-sODN and IgM into aggregated caps on the surface of human B cells. A role of surface Ig was furthermore shown by observations that antibody-mediated capping of B-cell surface IgM or IgD inhibited subsequent binding of sODNs and that the capacity of sODN to stimulate human B cells was blocked by excess IgM or IgG, by nonstimulatory antibodies to sIgM, as well as by a variety of negatively charged molecules. Together, these results indicate that sODNs engage surface Ig by charge-charge interactions that lead to activation of human B cells. PMID- 11902331 TI - Downmodulation of CD18 and CD86 on macrophages and VLA-4 on lymphocytes in experimental tuberculosis. AB - Development and evaluation of new vaccines and immunotherapy against tuberculosis demand a better understanding of the immune mechanisms in this disease. Costimulatory signals and intercellular contact seem to be pivotal in determining whether recognition of antigen by T cells leads to activation or anergy. In this paper, we show that virulent M. tuberculosis H37Rv downmodulates the ex vivo expression of CD18 and CD86 on peritoneal macrophages and VLA-4 on lymphocytes but does not disturb the in vitro production of interleukin (IL)-12 and interferon (IFN)-gamma after intraperitoneal infection. In addition, splenocytes from infected mice produce IL-10, while the expression of cell surface receptors is unchanged. The interplay among IL-12, IFN-gamma and IL-10 in vivo and the downmodulation of cell-surface receptors during the infection at the inflammatory site may contribute to the explanation of the maintenance of infection. PMID- 11902332 TI - Association of CD40 ligand expression on HTLV-I-infected T cells and maturation of dendritic cells. AB - Human T lymphotropic virus type I (HTLV-I) induces HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T-cell leukemia (ATL). The development of HAM/TSP is associated with rapid maturation of dendritic cells (DCs), while ATL is accomplished with their maturation defect. The DC maturation is induced by cell-to-cell contact with CD4+ T cells expressing CD40 ligand (L). We determined the influence of CD40L expressed on various HTLV-I infected T cells on the DC maturation. Around 60% of CD4+ T cells infected with HTLV-I for 1 week, expressed CD40L molecules involved in DC maturation. DCs matured by the CD40L+ T cells activated autologous CD4+ and CD8+ T cells. HTLV-I immortalized T-cell lines established from healthy donors consistently expressed CD40L molecules for 3 months, however, some lines lost the expression soon thereafter. Interleukin (IL)-2-independent and transformed lines lacked that expression. Furthermore, T cells obtained from HAM/TSP patients expressed CD40L molecules for at least 3 weeks, whereas T cells from ATL patients did not express that. The CD40L T cells did not induce DC maturation, and required exogenous CD40L molecules for maturation. The CD40L+ T-cell-induced maturation was blocked by anti-CD40L antibody. Therefore, the lack of CD40L expression on HTLV-I infected T cells may be associated with the development of ATL. PMID- 11902333 TI - Genesis of progressive T-cell deficiency owing to a single missense mutation in the common gamma chain gene. AB - Patients with a moderate X-linked combined immunodeficiency (XCID) owing to a single missense mutation in the common gamma chain (gammac) gene (L-->Q271) were found to have a progressive T-cell deficiency. Blood T cells from four older subjects with XCIDL-->Q271 were studied to ascertain the basis of that progression. Few CD4+ T cells displayed the phenotype (CD45RA+ CD62L+) or deletion circles from T-cell receptor (TCR) Vbeta-gene rearrangements found in recent thymic emigrants. These deficiencies were more severe in older males with XCIDL-->Q271. Relative frequencies of fresh CD4+ and CD8+ T cells that bound annexin V, an early indicator of programmed cell death, or propidium iodide, an indicator of cell necrosis, were greater in XCIDL-->Q271 T cells than in normal fresh T cells. The binding of annexin V and propidium iodide to XCIDL-Q271 T cells increased marginally after stimulation with anti-CD3, but binding by fresh or stimulated XCIDL-Q271 T cells exceeded that found in normal stimulated T cells. Also, telomeres from XCIDL-->Q271 CD4+ T cells were shortened in these patients compared to normal young adults. It therefore appears that the thymus is dysfunctional and that mature T cells are not effectively rescued from apoptosis or replication senescence via gamma-mediated pathways in XCIDL-->Q271. PMID- 11902334 TI - Blood gammadelta T cells and gammadelta TCR V gene specificities in a single missense mutation (L-->Q271) in the common gamma chain gene. AB - The numbers of blood CD4+, CD8+, and CD4-CD8- T cells bearing alphabeta T-cell receptor (TCR) or gammadelta TCR molecules in males with a single missense mutation (L-->Q271) in the common gamma chain gene (gamma(c)) were investigated by flow cytometry. Virtually all XCIDL-->Q271 blood T cells that were CD4+ or CD8+ displayed alphabeta TCR but no gammadelta TCR. In contrast, CD4-CD8- T cells from affected males usually displayed gammadelta TCR, but no alphabeta TCR. The gammadelta TCR specificities were also studied. Except for the oldest subject, there was a direct relationship between blood CD3+ T cells that displayed gammadelta TCR and Vgamma9 and Vdelta2a specificities. Relative frequencies of CD3+ blood T cells that were Vgamma9+ or Vdelta2a+ were inversely related to age. In the oldest patient, the only detected gammadelta TCR specificity was Vdelta1. Thus, in contrast to mice with no gamma(c), XCIDL-Q271 blood T cells that bear gammadelta TCR with Vgamma9/Vdelta2a specificities develop but then decline in late childhood and thereafter. TCR with the Vdelta1 specificity then appear in older survivors with XCIDL-->Q271. PMID- 11902335 TI - Responses of the rat immune system to arthritogenic adjuvant oil. AB - T-cell mediated inflammatory joint diseases with similarities to rheumatoid arthritis (RA) can be triggered in arthritis-prone rat strains by intradermal injection of adjuvant oils. The pathogenesis of oil-induced arthritis (OIA) remains elusive, and a largely unresolved question is how the rat immune system responds to arthritogenic oils such as incomplete Freund's adjuvant (IFA). Here we report that IFA already induces increased plasma levels of the acute-phase reactants (APR) fibrinogen and alpha1-acid glycoprotein at day 4 postinjection (p.i). In contrast, no early responses were detected in the joints before infiltration of the T cells, which coincided with arthritis onset at 11-14 days post injection (d.p.i.) The infiltrating cells were possibly derived from draining lymph nodes (LN), which were hyperplastic and contained increased cell numbers from 4 days p.i. and onwards. The magnitude of the early increase in cell numbers and APR was regulated by non-major histocompatibility complex (MHC) genes, as determined by comparison between arthritis-susceptible DA rats and arthritis-resistant but MHC-identical LEW.lAV1 and PVG.1AV1 rats. Arthritisprone DA rats developed a weak acute-phase response, suggesting that this systemic response may be counteracting disease. The DA rats also had the largest early increase in LN-cell numbers, suggesting that the LN hyperplasia is part of a disease pathway. The analysis of hyperplastic LN after in vivo labelling with bromodeoxyuridine (BrdU) revealed increased numbers and proportions of proliferating lymphocytes, including T cells. Furthermore, polymerase chain reaction (PCR)-analysis of LN cytokine mRNA revealed upregulation of interleukin (IL)-1beta at 4 d.p.i. We conclude that adjuvant oil exposure triggers both systemic acute phase reactions and local activation of the peripheral lymphoid system. These responses are genetically regulated and may determine arthritis development and susceptibility. PMID- 11902336 TI - Experimental IgG antibody production in vitro by peripheral blood and tonsil surface gamma+ B lymphocytes from Plasmodium falciparum-immune West Africans. AB - Antigen reactive B cells in tonsil specimens from teenagers from a region moderately exposed to P. falciparum were capable of being differentiated in vitro and producing specific immunoglobulin (Ig)G in up to 33% of individual experiments. Mononuclear cells or purified (s)gamma+ CD19+ B cells from peripheral blood or tonsil specimens from P falciparum-immune Senegalese subjects produced antigen-specific IgG upon appropriate stimulation in vitro. One fraction of this IgG was produced de novo by differentiated B cells and another fraction was likely bound on the surface of circulating or resident CD19+ sgamma+ B cells which were found in significantly greater numbers in individuals from rural Senegal as compared to nonimmune European controls. This study further documents the baseline levels of in vitro driven anti-P. falciparum IgG antibody production by mononuclear cells from blood and tonsils in immune populations exposed to P. falciparum differentially. Furthermore, this study demonstrates the relevance and potential utility of tonsils as a source of B lymphocytes to characterize further specific antibody responses to P. falciparum antigens in immune populations. PMID- 11902337 TI - Ontogeny of the murine Ig joining chain gene and protein. AB - We used Northern blot analysis in order to investigate the ontogeny of the murine joining (J)-chain gene. No J-chain expression was detected in embryonic tissues, including liver, spleen and intestine, but an expression of mu heavy chain was detected in foetal liver at day 17. J-chain expression was detected in the spleen at day 9 and in the intestine at day 15 after birth. Western blot analysis was carried out in order to compare the protein levels of J and mu heavy chains in serum from day 8 to day 24 after birth, using antihuman J chain and antimouse mu chain antibodies. Although mu chain protein could be detected in serum from day 8, J-chain protein was detectable only at day 24. These results suggest that the expression of J chain is a later event than the mu chain in the mouse, which thus differs in embryogenesis from humans. PMID- 11902338 TI - Stronger proliferative response to membrane versus cell-wall Streptococcal proteins by peripheral blood T cells in chronic plaque psoriasis. AB - Proliferative responses of peripheral blood mononuclear cells (PBMC) to group A streptococcal (GAS) antigens have been studied in 24 patients with psoriasis and 15 disease controls. Extracts of cell wall (including M protein) from types M4 and M12 GAS, recombinant M6 protein, and both cell-wall and cell-membrane extracts from type M6 (M6+) GAS and its corresponding M gene deletion mutant (M6 ) were tested. PBMC from psoriatic patients proliferated more strongly to cell wall extracts from M12 versus M4 (P = 0.0348), and to M6+ versus M6- (P = 0.0019) GAS with, in most cases, moderate proliferation to recombinant M6 protein. The psoriatic response to M12 cell wall was significantly increased compared to the controls (P = 0.0032). In psoriatics, M6+ membrane extracts induced a markedly greater proliferation than those of cell wall (P = 0.0002); responses to M6+ (P = 0.0039) and M6- (P = 0.0114) membrane extracts were higher than those of the control PBMC. Both groups showed a decreased response to the M6- versus M6+ membrane extracts (P = 0.0030; P = 0.0181, respectively). This study has demonstrated that patients with psoriasis have a heightened circulating T-cell response to cell wall M protein and to non-M proteins present on the cell wall and membrane of GAS. PMID- 11902339 TI - The effect of IgM-enriched human Ig and rabbit antithymocyte globulin on the stimulation of mononuclear cells. AB - Whether IgM-enriched intravenous Ig (pentaglobin) is a useful adjunct treatment for graft versus host disease (GvHD) prophylaxis in allogeneic stem-cell transplantation is unclear. Clinical data with the use of a five-agent GvHD prevention regimen, including pentaglobin and antithymocyte globulin (ATG), are encouraging. In vitro both have been reported to modulate alloreactive T cells. We compared their inhibitory effect on the phytohemagglutinin-induced lymphocyte proliferation. ATG blocked the proliferation of lymphocytes at lower doses and much stronger than pentaglobin. The combination of both was not different from ATG alone. In pentaglobin, glucose used as stabiliser, caused the effect. Starting at a concentration of 40 mg/dL glucose, glucose alone showed a dose dependent inhibition of phytohemaglutinin (PHA)-induced proliferation. For the in vivo application of pentaglobin, the results suggest that pentaglobin does not inhibit the proliferation of T cells. PMID- 11902341 TI - The patient.com, part 2. PMID- 11902340 TI - Increased expression of fas ligand in human tuberculosis and leprosy lesions: a potential novel mechanism of immune evasion in mycobacterial infection. AB - To study the location and mechanism of apoptosis within the human tuberculosis (TB) and leprosy lesions, parallel sections were analyzed for mycobacterial antigens (M.Ag), Fas ligand (FasL), Fas, CD68 and Mac387 by immunohistochemistry, and apoptotic cells by the terminal deoxynucleotidyl-transferase-mediated dUTP digoxigenin nick end labelling method. Cutaneous leishmaniasis and foreign body granulomas were analyzed for comparison. The heavily infected macrophages in multibacillary TB and leprosy granulomas very strongly expressed FasL, indicating that a mycobacterial infection can induce an increased expression of FasL in a population of infected macrophages, which may protect them from the attack of Fas expressing lymphocytes. However, macrophages with high levels of leishmania amastigotes did not selectively express FasL, suggesting that this phenomenon is specific for the mycobacteria. Interestingly, in the well-formed TB granulomas, 84% of the multinucleated giant cells strongly expressed FasL. The expression of Fas was weak (34%) or absent. A higher number (33%) of epithelioid cells expressed FasL than Fas (23%). Lymphocytes were scanty among the epithelioid cells. The frequency of apoptotic cells was higher in the epithelioid cells (0.25%) than the mononuclear cells in the mantle zone (0.14%). Thus, the epithelioid cells and the multinucleated giant cells by virtue of the increased expression of FasL may make these granulomas an immune privileged site for mycobacteria. PMID- 11902342 TI - Unscramble the alphabet soup of medicare payment systems. PMID- 11902343 TI - Eliminating the issue of skin color in assessment of the blanch response. AB - OBJECTIVE: The high melanin concentration in dark skin prevents the observation of a blanch response to light finger pressure. The objective of this study was to determine the ability of visible and near-infrared spectroscopy (the technique used in pulse oximetry) to monitor a blanch response from in vivo spectra in individuals with light and dark skin, based on changes in blood volume. DESIGN: A quasi-experimental repeated measures design was employed. A stepper motor with an attached spectrophotometer probe was used to deliver controlled pressure to the participants' forearms, mimicking the finger-blanching test. Visible and near infrared spectra were acquired throughout the blanching cycle. SETTING: The In Vivo Tissue Optics Lab at the Institute for Biodiagnostics, Winnipeg, Manitoba, Canada. PARTICIPANTS: A convenience sample of 10 healthy light-skinned individuals and 10 healthy dark-skinned individuals. RESULTS: Determined by analysis of the spectra, the 2 groups differed in pigmentation in both the visible (P<.01) and near-infrared (P<.01) regions of the absorbance spectrum. There was a significant difference in total hemoglobin at high and low pressure in both the visible (P<.01) and near-infrared (P<.05) regions. CONCLUSIONS: The observation of a significant difference in total hemoglobin at high and low pressure in both light- and dark-skinned groups in this study demonstrates the ability of visible and near-infrared spectroscopy to monitor blood volume changes associated with a blanch response. These findings support the potential use of this technology as the basis of a clinically useful blanch response tool that is insensitive to skin color. PMID- 11902344 TI - Comparison of 2 wound volume measurement methods. AB - OBJECTIVE: To compare 2 wound volume measurement techniques, the Kundin device and stereophotogrammetry, on 2 wound shapes. DESIGN: Using 2 wound measurement techniques, the interrater and intrarater reliability and the bias and standard error of measurement of an L-shaped and a pear-shaped plaster of paris wound model were assessed. SETTING: A clinical laboratory of a school of nursing. PARTICIPANTS: Twenty-four raters, all but 2 being registered nurses, measured each of the wounds using both techniques. INTERVENTIONS: Each rater measured each wound twice using each method in a randomly assigned order defined on a card that was drawn from a box. Measurements were recorded on a researcher-designed data collection form, which included some demographic data related to each participant. MAIN OUTCOME MEASURES: The study hypothesis was that there would be no significant difference in accuracy between the 2 wound volume measurement methods. RESULTS: The least biased and most accurate technique was stereophotogrammetry, with the smallest standard of error of measurement. Interrater reliability of average ratings was identical for both methods at 0.98. For single ratings, stereophotogrammetry was slightly higher than the Kundin device. Intrarater reliability was higher on the pear-shaped wound for the Kundin device, which had lower interrater reliability, suggesting that nurses were consistent in the direction and size of personal error. Intrarater reliability for stereophotogrammetry was identical to that of the Kundin device for the L shaped wound and lower for the pear-shaped wound. CONCLUSIONS: Although both techniques have acceptable accuracy, stereophotogrammetry is more accurate and has more clinical applications. PMID- 11902345 TI - Developing an interdisciplinary evidence-based skin care pathway for long-term care. PMID- 11902347 TI - Surveillance of mortality during a refugee crisis--Guinea, January-May 2001. AB - Since 1990, the republic of Guinea (2000 population: 7.5 million) has accepted 390,000-450,000 refugees from Sierra Leone and Liberia. During this 10-year period, refugees have lived in small villages scattered throughout rural southeastern Guinea. During September-December 2000, attacks by armed factions in Guinea led to the widespread displacement of refugees living in the southeastern camps; the refugees subsequently were transferred to safer camps in the northwest. Approximately 280,000 refugees initially were estimated to have been displaced. After the attacks, the number of refugees relocated was approximately 58,000. This report demonstrates methods used to calculate mortality rates when large populations are displaced. The findings indicate that the number of refugees in Guinea before the relocation probably was overestimated. The mortality rates calculated using conservative denominator numbers did not meet the definition of an emergency phase of a complex emergency, and mortality rates were lower for refugees compared with baseline rates for the local population. Accurate methods are needed to estimate population size in complex emergencies to provide resources to vulnerable groups. PMID- 11902346 TI - Pressure ulcers in America: prevalence, incidence, and implications for the future. An executive summary of the National Pressure Ulcer Advisory Panel monograph. PMID- 11902348 TI - Imported wild poliovirus causing poliomyelitis--Bulgaria, 2001. AB - In March 2001, a 13-month-old unvaccinated Roma (i.e., gypsy) girl from Bourgas, Bulgaria, had onset of bilateral leg weakness. The National Enterovirus Laboratory in the capital city of Sofia subsequently isolated a wild type 1 poliovirus in the patient's stool. In April, a second case, with wild type 1 poliovirus isolate was found in lambol located approximately 50 mileswest of Bourgas in an unvaccinated 26-month-old Roma girl who had onset of paralysis of both legs. Subsequent analyses indicated that these viruses were related closely to a strain isolated from Uttar Pradesh, India, in July 2000. A third confirmed case with clinical and serologic evidence of poliomyelitis was diagnosed in a 3 month-old Roma boy in Bourgas who had onset of paralysis on May 7. Following the identification of the poliovirus, the Bulgarian Ministry of Health implemented contact investigations, screening of children at high risk, retrospective record review, intensified acute flaccid paralysis (AFP) surveillance, and mass vaccinations. This report summarizes the outbreak investigation and supplemental vaccination activities in response to these polio cases. High routine vaccination coverage and certification standard AFP surveillance are necessary to detect rapidly and prevent the spread of poliovirus importations in areas and countries where polio is not endemic. PMID- 11902349 TI - Education, performance, quality and the march of technology. PMID- 11902350 TI - Effects of low-level laser therapy on malignant cells: in vitro study. AB - The aim of this study was to assess the effect of 635- and 670-nm laser irradiation on H.Ep.2 cells in vitro using MTT. In addition to our previous report on the effects of LLLT on the proliferation of laryngeal carcinoma cells in which it was found that irradiaton H.Ep.2 cells with 670-nm laser results in increased cell proliferation, it was decided to evaluate the effect of increased doses of laser light on these cells. The cells, obtained from SCC of the larynx, were routinely processed from defrost to the experimental condition. The cultures were kept either at 5% or 10% of FBS. Twenty-four hours after transplantation, the cells were irradiated with laser light (5-mW diode lasers; 635 and 670-nm; beam cross section approximately 1 mm) at local light doses between 0.04 and 4.8.10(4) Jm(-2). For 670 nm, significant differences in the proliferation were observed between the two concentrations of FBS (p = 0.002) and between irradiated cultures and controls (p = 0.000). Although the results were not significant, 635 nm irradiated cells also proliferated more than nonirradiated ones. This occurred under both conditions of nutrition. It is concluded, that irradiation with 670-nm laser light applied at doses between 0.04 and 4.810(4) Jm(-2) could significantly increase proliferation of laryngeal cancer cells. PMID- 11902351 TI - Er:YAG laser effects on enamel occlusal fissures: an in vitro study. AB - This study evaluated by scanning electron microscopy (SEM) the morphological changes in occlusal fissure enamel, of permanent models, irradiated by Er:YAG laser using contact and noncontact fiberoptics in vitro. Previous studies have demonstrated the efficacy of Er:YAG laser for dental hard tissue removal and cavity preparation. The treatment of occlusal fissures in noncarious permanent human molars (n = 9) was carried out with Er:YAG laser (KEY Laser II) using handpiece no. 2051, noncontact, focused (12 mm), water spray-cooled, pulse energy 200 mJ, and frequency 2 Hz (group 1), and handpiece no. 2055 with a quartz fiberoptic 50/10, in contact, air cooled, pulse energy setting of 350 mJ and frequency 2 Hz (group 2) and 400 mJ/2 Hz (group 3). The specimens were sectioned, dehydrated in a graded series of aqueous ethanol, dried, and sputtering with gold. Morphological change analysis on occlusal fissures was performed by SEM. Group 1 showed removal of fissure debris and predominantly enamel etching-like patterns, and groups 2 and 3 showed irregular edges, melting, and recrystallization of fissure enamel, with a lava-like structure and bubble-like voids. The results of this in vitro study suggest that the irradiation of fissures by Er:YAG laser using a fiberoptics (contact and air cooled) produced melting and recrystallization of fissures enamel. Further studies are required with different energy parameters and water cooling to evaluate the thermal effects on teeth. PMID- 11902352 TI - An update on photodynamic therapy applications. AB - Photodynamic therapy (PDT), following health agency approvals throughout the world for various cancers and other diseases, is slowly being accepted as a standard treatment to be added to the medical practitioner's armamentarium. This includes palliative treatments such as for obstructive esophageal and lung cancers as well as those intended for cure, early stage lung cancer, and actinic keratoses. A particularly new and important application is for the treatment of age-related macular degeneration (AMD), where PDT (Visudyne) has made a major impact on the outcome of this disease, the major cause of blindness in those over the age of 50. In the cancer field, while not yet approved (pending), the use of PDT in treatment of high-grade dysplasia (HGD) in Barrett's esophagus may well change how this disease is currently treated (often esophagectomy). Mechanistically, the recognition of apoptosis as an important mode of cell death following PDT and the critical role of the inflammatory process and immunity has only recently been recognized. This review updates the current and future role of PDT in cancer and other diseases. PMID- 11902353 TI - Effects of 1047-nm neodymium laser radiation on skin wound healing. AB - Previous research in our laboratory has shown that the polarization component of the electrical field plays an important role on the healing process of inflammatory lesions created in the end of the spinal column of Lewis rats, using a He-Ne laser at lambda = 632.8 nm. It is well known that polarization is lost in a turbid medium, such as living tissue. However, the Nd:YLF wavelength (lambda = 1,047 nm) allows more polarization preservation than lambda = 632.8 nm, and the Nd:YLF laser beam has been used in clinical trials as a biostimulating agent. In this work, we investigated the influence of a low-intensity, linearly polarized Nd:YLF laser beam on skin wound healing, considering two orthogonal directions of polarization. We have considered a preferential axis as the animals' spinal column, and we aligned the linear laser polarization first parallel, then perpendicular to this direction. Burns of about 6 mm in diameter were created with liquid N2 on the back of the animals, and the lesions were irradiated on days 3, 7, 10, and 14 postwounding, D = 1.0 J/cm2. Lesions 1 and 2 were illuminated using Nd:YLF pulsed laser radiation. Lesion 1 was irradiated with linear polarization parallel with the rat spinal column. Lesion 2 was irradiated using the same protocol, but the light polarization was aligned with the perpendicular relative orientation. Control lesions were not irradiated. We have taken photographs from the wound areas on the 3rd, 7th, 10th, 14th, and 17th postoperative day for a biometrical analysis. The results have shown that lesion 1 healed faster than the control lesions (p < 0,05), which presented a smaller degree of healing after 14 days postwounding. PMID- 11902354 TI - Laser literature watch. PMID- 11902355 TI - Lumbar percutaneous KTP532 wavelength laser disc decompression and disc ablation in the management of discogenic pain. AB - The objective of this research was to determine the outcome of laser disc decompression and laser disc ablation in the management of painful degenerative disc disease with or without associated disc prolapse. Nonendoscopic percutaneous laser disc decompression was performed under x-ray control via the posterolateral approach with side-firing probes. All patients with chronic back pain who had reproduced pain during discography of a nature, pattern, and distribution similar to what they experienced normally were included in the study. Magnetic resonance which confirmed stenosis and sequestrated discs, and patients with acute neurological findings were excluded from the study. Laser disc decompression or ablation was done using the KTP532 wavelength. The functional outcome was assessed prospectively using the Oswestry Disability Index. Clinical benefit was considered significant in those patients with a percentage change in the index of > or =50% at review 3-9 years (mean, 5.33 years) following surgery. A total of 52% of patients demonstrated a sustained significant clinical benefit, with an additional 21% in whom functional improvement was noted. Cohort integrity was 67%. Long-term benefit of the laser disc ablation and decompression for discogenic pain suggests a mechanism other than principally mechanical as a cause of chronic back and sciatic pain. It may suggest that efficacy occurs by reduction in the intradiscal production of irritative products and by an effect upon discal and annular neoneuralization. The sustained nature of the benefit after long-term preoperative symptoms (mean, 4.7 years) rules out any placebo effect. Selection should be restricted to patients without significant lateral recess stenosis, retrolisthesis or olisthesis of > or =3 mm, significant dorsal or foraminal osteophytosis, extrusion, or sequestration. PMID- 11902356 TI - Pressure ulcer surface area measurement using instant full-scale photography and transparency tracings. AB - BACKGROUND AND PURPOSE: An essential element in pressure ulcer management is the assessment of wound healing through measurement. The purpose of this study was to investigate the intraobserver and interobserver reliability of measuring pressure ulcers using instant full-scale photography combined with transparency tracings. SUBJECTS: 26 patients in 3 long-term-care facilities. METHODS: Duplicate photographs of 30 wounds from 26 subjects were obtained once a week over a period of 2 weeks, resulting in 120 photographs. Duplicate tracings of the photographs were subsequently assessed by 2 independent observers, resulting in 480 observations. The Intraclass Correlation Coefficient (ICC) was used as an indicator of chance-corrected agreement to estimate the relative reliability for the interobserver and intraobserver data. An Altman-Bland plot was also constructed to measure the relationship between interobserver differences and wound surface area. RESULTS: Analysis of the data showed that all measurement comparisons were highly reliable (ICCs = 0.99). No statistical differences between observed surface areas could be demonstrated. CONCLUSIONS: The combined wound measurement method described in this study represents a simple, practical, and inexpensive technique to accurately monitor and evaluate pressure ulcer healing. An instant full-scale photographic technique combined with transparency tracings of a wound should be considered for wound measurement, rather than each technique used independently. The results of this study indicate that measurements obtained with this combined method are highly reliable within and between observers. PMID- 11902357 TI - Maintenance of skin integrity as a clinical indicator of nursing care. AB - OBJECTIVE: To assess maintenance of skin integrity in hospitalized patients as a clinical indicator of quality nursing care. DESIGN: Descriptive correlational study. SETTING: 17 acute care urban and rural hospitals in Texas. PARTICIPANTS: 723 hospitalized patients from 33 medical-surgical units. MAIN OUTCOME MEASURES: Pressure ulcers since admission and nursing care practices per unit. MAIN RESULTS: Overall prevalence of pressure ulcers since admission (4.7%) was lower than previously reported for acute care settings. Prevalence of pressure ulcers in hospitalized patients (10%) was higher than found in the Texas Nurses Association feasibility study (3.7%). The skin integrity ratio was strongly correlated for several unit variables, including number of beds per unit (r = 0.623) and average daily census per unit (r = 0.909). Benchmarking data across units showed that units with subjects that maintained skin integrity had a lower percentage of patients assessed on admission and performed daily assessments less frequently than did the units with subjects that did not maintain skin integrity. In addition, the units with subjects that maintained skin integrity classified fewer patients as at risk for pressure ulcer development and did not implement a skin care protocol for these patients. CONCLUSION: Patients who developed a pressure ulcer after admission were older and had more risk for pressure ulcers than those who maintained skin integrity. Benchmarking data detected various differences in nursing care. Significant relationships between study variables demonstrate the importance of assessing clinical indicators to monitor nursing care. Outcomes such as skin integrity, pressure ulcer since admission, and nosocomial ratio represent the quality of nursing care. PMID- 11902358 TI - Customer service: an important leadership skill for every ADA member. PMID- 11902359 TI - The key to third party reimbursement? Establish a dialogue with physicians on their level. PMID- 11902360 TI - What roles should RDs be "claiming" when dealing with health claims? PMID- 11902362 TI - Developmental training areas for upskilling. PMID- 11902363 TI - Do dietetics professionals need to be concerned with dress on the job? PMID- 11902361 TI - Client satisfaction: turning referrals into regulars. PMID- 11902364 TI - Folic acid fortification: additional issues. PMID- 11902365 TI - Nutrition programs to benefit from election-year politics? PMID- 11902366 TI - Thirty years of the Older Americans Nutrition Program. PMID- 11902367 TI - Sugar and sugars: myths and realities. PMID- 11902368 TI - Perceptions and practices of self-defined current vegetarian, former vegetarian, and nonvegetarian women. AB - OBJECTIVE: To assess the diversity of vegetarians' dietary practices and how they change over time, and to explore perceptions of meat and dairy products among vegetarians, former vegetarians, and nonvegetarians. DESIGN: Cross-sectional survey; qualitative interviews with a subsample. SUBJECTS/SETTING: Ninety self defined current vegetarian, 35 former vegetarian and 68 nonvegetarian women in Vancouver, British Columbia. A subsample of 15 subjects completed qualitative interviews. STATISTICAL ANALYSIS PERFORMED: Group comparisons using 1-way analysis of variance with post-hoc testing for continuous variables, chi2 for categorical variables. RESULTS: Of 90 current vegetarians, 51 and 14 reported occasional use of fish or chicken respectively. Fifty-six vegetarians, including 4 of 6 vegans, reported that their diets had become more restrictive over time, and 48 planned additional changes, most frequently a reduction in dairy product use. Reasons cited by former vegetarians for resuming omnivorous diets included: not feeling healthy, concern about their nutritional status, a change in living situation, or missing the taste of meat. Perceptions of meat and dairy products differed significantly by dietary pattern: nonvegetarians and former vegetarians were more likely than current vegetarians to agree with statements inferring positive attributes (eg, nutrient content). In contrast, more current and former vegetarians than nonvegetarians agreed with statements inferring negative attributes (eg, presence of contaminants). APPLICATIONS/CONCLUSIONS: Dietitians who counsel women need to be aware of the heterogeneity of dietary practices and beliefs regarding use of animal products to provide advice appropriate to each individual. At a broader level, addressing women's food safety and animal welfare concerns will likely require collaboration among food industry and government, health, and consumer agencies. PMID- 11902369 TI - Feeding dysfunction is associated with poor growth and health status in children with cerebral palsy. AB - OBJECTIVE: To describe parent-reported feeding dysfunction and its association with health and nutritional status in children with cerebral palsy. DESIGN: Anthropometry was measured and z scores calculated. The Child Health Questionnaire was used to assess health status, and a categorical scale (none to severe) was used to classify subjects according to severity of feeding dysfunction. SUBJECTS: 230 children (9.7+/-4.6 years; 59% boys) with moderate to severe cerebral palsy were recruited from 6 centers in the United States and Canada. STATISTICAL ANALYSES: Descriptive statistics, the Kruskal-Wallis and Pearson chi2 tests. RESULTS: Severity of feeding dysfunction was strongly associated with indicators of poor health and nutritional status. The mean weight z scores were -1.7, -2.5, -3.3, and -1.8 among children with none, mild, moderate, or severe (largely tube-fed) feeding dysfunction, respectively (P= .003). Similar results were observed for height z score (P=.008), triceps z score (P=.03), and poor Global Health score (part of the Child Health Questionnaire) (P<.001). Subjects who were tube fed were taller (P=.014) and had greater body fat stores (triceps z score, P=.001) than orally fed subjects with similar motor impairment. For subjects exclusively fed by mouth, a dose-response relationship was observed between feeding dysfunction severity and poor nutritional status. Subjects with only mild feeding dysfunction had reduced triceps z score (-0.9) compared with those with no feeding problems (-0.3). CONCLUSION: For children with moderate to severe cerebral palsy, feeding dysfunction is a common problem associated with poor health and nutritional status. Even children with only mild feeding dysfunction, requiring chopped or mashed foods, may be at risk for poor nutritional status. Parental report of feeding dysfunction with a structured questionnaire may be useful in screening children for nutritional risk. PMID- 11902371 TI - The food intake recording software system is valid among fourth-grade children. AB - OBJECTIVES: To assess the validity of the Food Intake Recording Software System (FIRSSt) against observation of school lunch and a 24-hour dietary recall (24hDR); and to test the effects of sequencing, observation and a hair sample as a bogus pipeline on accuracy of dietary report. DESIGN: Six-group design systematically varying sequence of self-report (FIRSSt vs 24hDR), observation of school lunch and hair sample as a bogus pipeline manipulation, with random assignment of participants. SUBJECTS/SETTING: 138 fourth-grade students in 2 elementary schools. MAIN OUTCOME MEASURES: Accuracy of reported food consumption was measured in terms of matches, intrusions, and omissions among the FIRSSt, 24hDR, and as observed at school lunch. Students also completed self-report of performance with FIRSSt. STATISTICAL ANALYSIS PERFORMED: t tests, Pearson correlations, analysis of variance, factor analysis. RESULTS: When compared with school lunch observation for one meal, FIRSSt attained 46% match, 24% intrusion and 30% omission rates, while a dietitian-conducted 24hDR obtained 59% match, 17% intrusion, and 24% omission rates. FIRSSt attained 60% match, 15% intrusion, and 24% omission rates against 24hDR for all meals in the previous day. There was no evidence of sequence of assessment affecting accuracy indicators, but there was a weak effect of school lunch observation on percent intrusions. Obtaining a hair sample reduced the omission rate for FIRSSt vs 24hDR and increased the match rate for 24hDR vs observation, thereby enhancing this as a bogus pipeline procedure. Children generally enjoyed completing FIRSSt. Hispanic children were more likely to report problems using FIRSSt. APPLICATIONS/CONCLUSIONS: FIRSSt is somewhat less accurate than a dietitian-conducted 24hDR. However, this lower-cost procedure provides a promising method for assessing diet among children. Observation of consumption at school lunch may be reactive and artificially increase agreement. Obtaining a hair sample as a bogus pipeline may be a valuable technique for enhancing the accuracy of dietary assessment among children. PMID- 11902370 TI - Nutritional status of teenage female competitive figure skaters. AB - OBJECTIVE: To assess the nutritional status of female competitive figure skaters during preseason, competitive season, and off-season. SUBJECTS: Eighteen female competitive figure skaters, age range 14 to 16 years, from the New England region. STATISTICS: Data was analyzed by repeated measures analysis of variance, Duncan Multiple Range Tests, one-sample t tests, and confidence intervals. DESIGN: Nutrient intakes were determined from 3-day diet records. Body composition was assessed through heights, weights, and underwater weighing. Blood samples were drawn for analysis of selected indexes of nutritional status. RESULTS: Height and weight did not differ significantly among the seasons. Body fat was 1.1 kg higher off-season compared with preseason. Energy intake over the 3 seasons did not vary significantly (mean preseason: 1,678 kcal/day; competitive season, 1,630 kcal/day; off-season: 1,673 kcal/day) (P>.05). During the competitive season 78%, 50%, and 44% of the skaters had intakes less than 67% of RDA for folate, iron, and calcium, respectively. Most of the biochemical indexes of nutritional status were within normal limits. CONCLUSIONS: The results indicate that teenage female skaters have relatively low energy intake and inadequate intakes of certain nutrients, which may account for some of the observed seasonal variations in blood markers of nutritional status. These findings point to the need for nutrition education for these athletes, especially during their competitive season when nutritional status may be compromised. PMID- 11902372 TI - Medical costs are reduced when children with intractable epilepsy are successfully treated with the ketogenic diet. AB - The ketogenic diet is used for children with drug refractory epilepsies. Although this diet was developed early in the 20th century, its use was infrequent until recently. One of the concerns about the ketogenic diet is the cost of initiating the diet. The process generally involves an inpatient visit that can last several days. In this study, the health care costs for 15 children with drug refractory epilepsy who received their care at Vanderbilt University Medical Center were compared for the period of time 6 to 12 months before they began the ketogenic diet and 6 to 12 months after they began the ketogenic diet. All comparisons within each child were done on the same amount of time before and after initiation of the diet. Total costs affiliated with care for the 15 children were: $352,820.20 for the prediet period, $41,221.91 for the diet initiation, and $149,436.86 for the postdiet-initiation period. We conclude that successful maintenance on the ketogenic diet provides a substantial financial benefit. PMID- 11902373 TI - The history of enteral nutrition therapy: from raw eggs and nasal tubes to purified amino acids and early postoperative jejunal delivery. AB - Although enteral feeding therapy has existed since ancient Egypt, most of the major advances in enteral feeding techniques and formulas took place during the 20th century, including postpyloric tube placement in 1910; continuous and controlled delivery of liquid nutrition in 1916; feeding during surgery and modification of macronutrients in 1918; feeding via a pump in 1930; recognition of the importance of nutrition therapy during injury recovery and the addition of micronutrients and early postoperative feeding in 1940; the introduction of commercial products during the 1950s with chemically defined formulas following a decade later; and the development of modern formulas during the 1970s. The purpose of this review is to provide a historical account of enteral nutrition, including modes and routes of delivery, types of diet, and refinements in delivery techniques and formulas and to offer the history of the therapy as a resource for developing and improving enteral feeding techniques and therapies and implementing optimal patient care strategies. PMID- 11902374 TI - Growth retardation in children with epilepsy on the ketogenic diet: a retrospective chart review. PMID- 11902375 TI - Foods on students' trays when they leave the cafeteria line as a proxy for foods eaten at lunch in a school-based study. PMID- 11902376 TI - Food choices of third-grade children in Texas. PMID- 11902377 TI - Prevalence of nonvitamin, nonmineral supplement usage among university students. PMID- 11902378 TI - Glycemic index of popular sport drinks and energy foods. PMID- 11902379 TI - Nutrition knowledge of collegiate athletes in a Division I National Collegiate Athletic Association institution. PMID- 11902380 TI - The relationship between health beliefs and behaviors and dietary intake in early adolescence. PMID- 11902381 TI - Is nutritional diagnosing a critical step in the nutrition care process? PMID- 11902382 TI - Effects of nutrition education programs on anthropometric measurements and pregnancy outcomes of adolescents. PMID- 11902383 TI - California Project LEAN's Food on the Run program: an evaluation of a high school based student advocacy nutrition and physical activity program. PMID- 11902384 TI - Assessment of nutritional status and motivation to make behavior changes among adolescents. PMID- 11902385 TI - Adolescent growth and development. PMID- 11902386 TI - Adolescent psychosocial development. PMID- 11902387 TI - Strategies for counseling adolescents. PMID- 11902388 TI - Individual and environmental influences on adolescent eating behaviors. AB - Food choices of adolescents are not consistent with the Dietary Guidelines for Americans. Food intakes tend to be low in fruits, vegetables, and calcium-rich foods and high in fat. Skipping meals is also a concern among adolescents, especially girls. Factors influencing eating behaviors of adolescents need to be better understood to develop effective nutrition interventions to change eating behaviors. This article presents a conceptual model based on social cognitive theory and an ecological perspective for understanding factors that influence adolescent eating behaviors and food choices. In this model, adolescent eating behavior is conceptualized as a function of individual and environmental influences. Four levels of influence are described: individual or intrapersonal influences (eg, psychosocial, biological); social environmental or interpersonal (eg, family and peers); physical environmental or community settings (eg, schools, fast food outlets, convenience stores); and macrosystem or societal (eg, mass media, marketing and advertising, social and cultural norms). PMID- 11902389 TI - Designing effective nutrition interventions for adolescents. AB - By altering dietary behaviors, nutrition interventions during adolescence have the potential of affecting children at that time and later in life. The majority of interventions implemented in the teen years have occurred in schools, but other intervention sites have included after-school programs, summer camps, community centers, libraries, and grocery stores. Programs with successful outcomes have tended to be behaviorally based, using theories for the developmental framework; included an environmental component; delivered an adequate number of lessons; and emphasized developmentally appropriate strategies. One planning method that can be used in the development of nutrition interventions is Intervention Mapping. The steps of Intervention Mapping include conducting a needs assessment, developing proximal program objectives, mapping appropriate strategies and methods to address the objectives, planning the program design, planning program adoption and implementation, and evaluation. The use of intervention-planning techniques, coordination of nutrition and physical education interventions, using technological advances such as CD-ROMs, incorporation of policy changes into intervention efforts, and dissemination of effective programs are all trends that will influence the future development of effective nutrition programs for adolescents. PMID- 11902390 TI - HealthWorks! Weight management program for children and adolescents. PMID- 11902391 TI - Body basics: a nutrition education program for adolescents about food, nutrition, growth, body image, and weight control. PMID- 11902392 TI - Promoting a healthful lifestyle and encouraging advocacy among university and high school students. PMID- 11902393 TI - Clueless in the mall: a web site on calcium for teens. PMID- 11902394 TI - An environmental intervention to improve a la carte foods at middle schools. PMID- 11902395 TI - Impact of an educational seminar on high school students' knowledge of folic acid supplementation and its role in the prevention of birth defects. PMID- 11902396 TI - Nutritional issues for adolescents. AB - This article describes important issues for the nutritional health of adolescents. The issues discussed include dietary factors involved in cardiovascular and cancer disease risk; osteoporosis and bone mineralization; overweight and obesity; related risk factors such as type 2 diabetes; and eating disorders. The discussion focuses on nutritional issues for the general adolescent population, rather than high-risk adolescents. Data are primarily drawn from large, population-based studies using representative samples of adolescents. PMID- 11902397 TI - The great beginnings program: impact of a nutrition curriculum on nutrition knowledge, diet quality, and birth outcomes in pregnant and parenting teens. PMID- 11902398 TI - The right bite program: a theory-based nutrition intervention at a minority college campus. PMID- 11902400 TI - Integrating the Food Guide Pyramid and Physical Activity Pyramid for positive dietary and physical activity behaviors in adolescents. PMID- 11902399 TI - Gimme 5: an innovative, school-based nutrition intervention for high school students. PMID- 11902401 TI - Youth tobacco surveillance--United States, 2000. AB - PROBLEM/CONDITION: Tobacco use is the single leading preventable cause of death in the United States, accounting for approximately 430,000 deaths each year. The prevalence of cigarette smoking nationwide among high school students increased during the 1990s, peaking during 1996-1997, then began a gradual decline. Approximately 80% of tobacco users initiate use before age 18 years. If the trend in early initiation of cigarette smoking continues, approximately 5 million children aged <18 years who are living today will die prematurely because they began to smoke cigarettes during adolescence. The economic costs associated with tobacco use ranges from $53 billion to $73 billion per year in medical expenses and $47 billion in lost productivity. Because of these health and economic consequences, CDC has recommended that states establish and maintain comprehensive tobacco-control programs to reduce tobacco use among youth. REPORTING PERIOD: January 2000 through December 2000. DESCRIPTION OF THE SYSTEM: To assist states in developing and maintaining their state-based comprehensive tobacco prevention and control programs, CDC developed the Youth Tobacco Surveillance and Evaluation System, which includes international, national, and state school-based surveys of middle school and high school students. Two components of this system are discussed in this report--the National Youth Tobacco Survey and the state Youth Tobacco Surveys. The national survey is representative of students in the 50 states and the District of Columbia; 35,828 students in 324 schools completed questionnaires in the spring of 2000. Twenty nine state surveys were conducted in the spring and fall of 2000; state sample sizes ranged from 583 to 33,586 students. This report summarizes data from the 2000 national survey and state surveys. RESULTS AND INTERPRETATION: Findings from the National Youth Tobacco Survey indicate that current tobacco use ranges from 15.1% among middle school students to 34.5% among high school students. Cigarette smoking is the most prevalent form of tobacco use, followed by cigar smoking and smokeless tobacco use. Approximately one half of current cigarette smokers in middle school and high school report that they usually smoke Marlboro cigarettes. Black students are more likely to smoke Newport cigarettes than any other brand. More than one half of current cigarette smokers in middle school and high school report that they want to stop smoking. Nearly one fourth of middle school and high school students who have never smoked cigarettes are susceptible to initiating cigarette smoking in the next year. Exposure to secondhand smoke (e.g., environmental tobacco smoke) is substantially higher among both middle school and high school students. During the week before the survey, approximately 9 out of 10 current cigarette smokers and one half of never cigarette smokers were in the same room with someone who was smoking cigarettes; and 8 out of 10 current cigarette smokers and 3 out of 10 never cigarette smokers rode in a car with someone who was smoking cigarettes. Approximately 70% of middle school and 57% of high school students who currently smoke cigarettes live in a home where someone smokes cigarettes. Among never cigarette smokers, approximately 3 out of 10 live in a home where someone smokes cigarettes. Approximately 69% of middle school and 58% of high school students aged <18 years who currently smoke cigarettes were not asked to show proof of age when they bought or tried to buy cigarettes. Approximately 8 out of 10 middle school and high school students have seen antismoking commercials. Eight out of 10 middle school students report having seen actors using tobacco on television or in the movies, and approximately 11% of middle school and 16% of high school students who had never used tobacco would wear or use something with a tobacco company name or picture on it. This rate increases to nearly 60% for current tobacco users. PUBLIC HEALTH ACTIONS: Youth Tobacco Survey data are used by health and education officials to improve national and state programs to prevent and control youth tobacco use. Several states use the data in presentations to their state legislators to demonstrate the need for funding smoking cessation and prevention programs for youth. PMID- 11902402 TI - Quantitative trait locus mapping in chickens by selective DNA pooling with dinucleotide microsatellite markers by using purified DNA and fresh or frozen red blood cells as applied to marker-assisted selection. AB - Many large, half-sib sire families are an integral component of chicken genetic improvement programs. These family structures include a sufficient number of individuals for mapping quantitative trait loci (QTL) at high statistical power. However, realizing this statistical power through individual or selective genotyping is yet too costly to be feasible under current genotyping methodologies. Genotyping costs can be greatly reduced through selective DNA pooling, involving densitometric estimates of marker allele frequencies in pooled DNA samples. When using dinucleotide microsatellite markers, however, such estimates are often confounded by overlapping "shadow" bands and can be confounded further by differential amplification of alleles. In the present study a shadow correction procedure provided accurate densitometric estimates of allele frequency for dinucleotide microsatellite markers in pools made from chicken purified DNA samples, fresh blood samples, and frozen-thawed blood samples. In a retrospective study, selective DNA pooling with thawed blood samples successfully identified two QTL previously shown by selective genotyping to affect resistance in chickens to Marek's disease. It is proposed that use of selective DNA pooling can provide relatively low-cost mapping and use in marker-assisted selection of QTL that affect production traits in chickens. PMID- 11902403 TI - Genotype-by-environment interaction with broiler genotypes differing in growth rate. 3. Growth rate and water consumption of broiler progeny from weight selected versus nonselected parents under normal and high ambient temperatures. AB - One cycle of high-intensity selection on BW was conducted to study correlated effects on performance under high ambient temperature (AT). From a large flock of a commercial sire-line, 3 males and 15 females with the highest BW at 35 d of age were mated and produced a group of 120 BW-selected chicks. Three average-BW males and 15 average-BW females from the same flock were mated to produce a control group of 120 chicks. On Day 17, the two groups were equally divided between two temperature-controlled chambers and housed in individual cages. One chamber was set to a normal AT (NAT; constant 22 C) and the second chamber to high AT (HAT; constant 32 C). Under NAT, the relative advantage of the selected broilers over the controls did not change from 17 to 42 d of age, averaging about 15% for BW gain and 9.7% for feed consumption. These differences were halved under HAT from Days 17 to 28 and were reversed from 28 to 42 d of age, when the selected broilers consumed significantly less feed and gained less BW than the controls. Water-to-feed ratio was measured in each AT treatment. From 28 to 42 d of age, averaged over the two groups, birds under HAT consumed 2.5 g water/g of feed compared to only 1.5 g water/g feed under NAT. The diminished superiority of the selected broilers under HAT led to a substantial genotype-by-environment interaction involving high AT and within-stock genetic differences in growth rate. It appears that broilers selected for rapid growth under optimal conditions do not achieve their genetic potential under high AT. Thus, specific indicators of adaptation to heat, possibly water consumption or body temperature, should be added to commercial selection for rapid growth to improve broiler performance in hot climates. PMID- 11902404 TI - Genetic and phenotypic correlations between antibody responses to Escherichia coli, infectious bursa disease virus (IBDV), and Newcastle disease virus (NDV), in broiler lines selected on antibody response to Escherichia coli. AB - The genetic control of antibody (Ab) response to Escherichia coli (EC), infectious bursa disease virus, and Newcastle disease virus and the genetic and phenotypic correlation between these Ab responses, were evaluated under farm conditions in which chicks were simultaneously exposed to these antigens. The experimental population comprised five groups: two lines divergently selected for high (HH) or low (LL) Ab response to EC vaccination; a commercial broiler dam line (CC), from which HH and LL had been derived; and the HH x CC and LL x CC hybrid groups (HC and LC, respectively). Lines LL and HH expressed similar symmetric divergence to all three antigens. The ranking of the LL, LC, CC, HC, and HH genetic groups according to their mean Ab responses and their very high linear correlation with the LL vs. HH genomic scale clearly indicate the additive nature of the genetic divergence between these lines. Several estimates of correlation were calculated between Ab responses of each pair of antigens and between BW and Ab to each antigen. The high correlation between group means, the near-zero within-group correlation, and the low phenotypic correlation indicate the strongly positive genetic correlation between Ab responses and no correlation with BW. The results of this study suggest that overall immunocompetence of commercial broilers can be improved by selection for high Ab response of young chicks to controlled immunization with a single antigen, without counteracting further selection for high BW. PMID- 11902405 TI - Genetics of growth and reproduction in the turkey. 15. Effect of long-term selection for increased egg production on the genetics of growth and egg production traits. AB - A line (E) of turkeys selected long-term (37 generations) for increased egg production was reciprocally crossed with its randombred control population (RBC1) that served as the base population of the E line to study the influence of long term selection on the development of nonadditive genetic variation for egg production and body weight traits. Heterosis was significant for BW at 8, 16, and 20 wk of age and at 50% production (females only). At 16 and 20 wk of age, heterosis for BW was significant only for male offspring. No heterosis was observed in the reciprocal crosses for egg production when measured for 84, 180, or 250 d. Heterosis was significant for rate of response to stimulatory lighting of 14 h light per day (days from stimulatory lighting to production of first egg). Based on data for a 250-d production period, heterosis was observed in average clutch length but not in total days lost from broodiness or the effective length of the laying period (250-d lost in periods of 5 or more consecutive d at the end of the laying period). The present results suggest that long-term selection for increased egg production and the correlated decrease in BW increased the relative nonadditive genetic variation in BW. Reciprocal effects were significant for BW at 8 and 16 wk of age, probably due to a large difference in egg weight between the E and RBC1 lines. PMID- 11902406 TI - Fearfulness and performance related traits in selected lines of Japanese quail (Coturnix japonica). AB - Fearfulness and economic traits were studied in three lines of Japanese quail. Two of the lines were of the same genetic origin and were subjected to divergent selection for the duration of tonic immobility (TI), a measure of fearfulness. Birds were selected for long (LTI) or short (STI) duration of TI. The third line (DD) was of a different genetic origin and had been selected for early egg production. Fear, growth, residual feed intake, and measures of egg composition and production varied among lines. The distribution of TI in Line DD was closer to that from Line STI. Residual feed intake and shell content were lowest in the DD line. The DD birds laid more broken eggs than quail of the other lines. The STI line birds had higher BW and laid more, but smaller, eggs than LTI line birds. Eggs laid by LTI line birds had higher albumen content, but lower percentage shell, than those laid by STI line birds. When all traits were considered together, there was an overall tendency for STI line birds to out perform LTI birds with DD line birds showing intermediate performance. This finding supports the notion that there is a relationship between fearfulness and productivity. However, the skewed distribution of TI precluded estimation of correlation with production traits in the LTI and STI lines. No significant relationships among fear and production-related traits were found in the DD line, which contradicts the notion that fearfulness and production are related. PMID- 11902408 TI - Effect of nest design, passages, and hybrid on use of nest and production performance of layers in furnished cages. AB - Production performance, including egg quality, and proportion of eggs laid in nests were studied in furnished experimental cages incorporating nests, litter baths, and perches. The study comprised a total of 972 hens of two genotypes: Lohmann Selected Leghorn (LSL) and Hy-Line White. The birds were studied from 20 to 80 wk of age, and conventional four-hen cages were included as a reference. In furnished cages for six hens, the effects of 30 or 50% vs. 100% nest bottom lining (Astro turf) were studied with LSL hens. Nest bottom lining had no significant effect on egg production or proportions of cracked or dirty eggs, but the use of nests was significantly higher in cages incorporating nests with 100% lining, compared with 50 or 30%. The two hybrids were compared when housed in large, group-furnished cages for 14 or 16 hens of two designs; with a rear partition with two pop holes or fully open, i.e., no rear partition. LSL birds produced significantly better and had a significantly lower proportion of cracked eggs. There was no difference between H- and O-cages, either in production or in egg quality. LSL birds laid a significantly lower proportion of eggs in the nests, especially in O-cages, implying a significant hybrid x cage interaction. When housed in conventional cages, the hybrids did not differ in proportion of cracked eggs but differed in production traits. It was concluded that with the present nest design, the proportion of nest bottom lining cannot be reduced without affecting birds' use of nests, but the proportion did not affect exterior egg quality. The effect of genotype should be considered in the further development of furnished cages. PMID- 11902409 TI - Egg quality in furnished cages for laying hens--effects of crack reduction measures and hybrid. AB - Egg quality, egg production, and hen use of facilities, with respect mainly to proportion of eggs laid in nests and bird locations after lights-out, were studied in furnished-cage models for six or eight birds. In these cages, most of the eggs are laid in the 25 cm wide nest, i.e., egg laying is concentrated to a much smaller area compared with conventional cages. The study (20 to 80 wk of age) used 1,296 hens of three genotypes Hy-Line White, Hy-Line Brown, and Lohmann Selected Leghorn. Conventional four-hen cages were included as a reference. We separately studied the effects on the proportion of eggs cracked by egg saver wires in front of the egg cradle, long nest curtains, and position of nest opening (rear or front) with 30 or 100% of the nest bottom being lined with AstroTurf. Egg saver wires and long nest curtains lowered the proportions of cracked eggs significantly by softly catching and reducing the speed of the eggs before entering the egg cradle. The position of the nest opening affected none of the measured parameters. Less lining in the bottom of the nest, i.e., a reduction to 30% coverage, resulted in significantly higher proportions of dirty eggs and lower proportions of eggs laid in nests. Hybrid differences were found in most of the measured traits. We concluded that devices like egg savers and long nest curtains are effective measures in reducing cracks in furnished cages, where egg laying is concentrated to a much smaller area than in conventional cages. Reduction of the bottom lining to 30% makes nests less attractive to birds laying eggs. PMID- 11902407 TI - Effect of induced molting on albumen quality, hatchability, and chick body weight from broiler breeders. AB - Commercial Cobb broiler breeders were subjected to molting from 55 to 62 wk of age. Incubating eggs were collected before molting and after molting and were stored for 18, 13, 8, or 3 d. Sample eggs were broken to measure albumen Haugh units (HU) and pH. Two hundred twenty-five eggs per treatment, stored for 18, 13, 8, or 3 d, were incubated for 21 d; the hatched chicks were weighed at the end of incubation and again after Day 7 of rearing. As the storage time increased, albumen HU decreased (P < 0.001). At all storage times, HU after molting were higher than those before molting (P < 0.001). Albumen pH increased with storage time (P < 0.001). A molting x storage interaction on pH was observed after 8 d of storage (P = 0.03). Hatchability of eggs increased after hens were molted, if the eggs were stored for a long time (P < 0.001). Body weights of 1-d-old chicks from the eggs of hens before molt were heavier than those from eggs after molting (P < 0.001). Conversely, at 7 d, the chicks from the eggs after molting were significantly heavier than those from eggs before molting (P < 0.001). We concluded that molting was a procedure to improve hatchability and chick juvenile growth. If eggs need to be stored, we recommended that fresh eggs with high HU value be stored rather than those with low HU values. PMID- 11902410 TI - The use of diclazuril in extended withdrawal anticoccidial programs: 1. Efficacy against Eimeria spp. in broiler chickens in floor pens. AB - A 49-d floor pen study was conducted with broiler chickens to compare the effects of different anticoccidial withdrawal times on the efficacy of 1 ppm diclazuril. The starter diet in three treatments contained 66 ppm salinomycin + 50 ppm roxarsone (3-nitro-4-hydroxyphenylarsonic acid), followed by 1 ppm diclazuril in the grower diet commencing on Day 17. Diclazuril was withdrawn from these treatments on Day 28, 35, or 42 (finisher diet), respectively. Two other treatments in the study were given 66 ppm salinomycin + 50 ppm roxarsone in the starter and grower diets to Day 28 or no anticoccidial (unmedicated). The starter (Days 0 to 16), grower (Days 17 to 35), and finisher (Days 36 to 49) diets in each treatment included 55 ppm bacitracin methylene disalicylate for growth promotion. Fifty 1-d-old chicks were randomly allotted to each of 10 pens per treatment using a randomized complete block design. Birds in each pen were raised on litter naturally contaminated with a mixture of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. The results demonstrated that some performance loss occurred in the salinomycin Day 28 treatment. Means for weight gain and feed conversion on Day 49 were improved (P < 0.05) in each diclazuril treatment in comparison with the salinomycin and unmedicated treatments. Feed conversion means in the diclazuril Day 35 and Day 42 treatments were improved (P < 0.05) in comparison with the diclazuril Day 28 treatment, indicating that shorter withdrawal times provided further benefit. PMID- 11902411 TI - The use of diclazuril in extended withdrawal anticoccidial programs: 2. Immunity to Eimeria tenella challenge after drug withdrawal. AB - The effect of diclazuril medication on the development of natural immunity to Eimeria tenella was determined. Birds in two treatments, nonexposed, non challenged (NENC) and nonexposed, challenged (NEC), were from a holding group raised under conditions designed to prevent coccidial infection. Birds in five other treatments, from the same hatch and source as birds in the first two treatments, were from a floor pen study involving a natural exposure to Eimeria spp. These birds were assigned to the current study based on their respective treatments in the original floor pen study as follows: unmedicated (UNM), 66 ppm salinomycin (SAL) + 50 ppm roxarsone in the starter and grower diets to Day 28 (SAL 28), 66 ppm SAL + 50 ppm roxarsone in the starter diet and 1 ppm diclazuril (DIC) in the grower diet to Day 28 (DIC 28) or to Day 35 (DIC 35), or in the grower and finisher diets to Day 42 (DIC 42). Each treatment comprised three floor pens of 10 birds (female) in a randomized complete block design. All birds were fed an UNM finisher diet during the 7-d challenge study. Birds in each treatment, except NENC, were individually inoculated (p.o.) with 1 x 10(5) Eimeria tenella sporulated oocysts on Day 1. Based on bird performance and cecal lesion scores, birds in the DIC 35 and 42 treatments had a low immunity to the challenge infection, birds in the DIC 28 treatment were partially immunized, and the highest levels of immunity were observed in the UNM and SAL 28 treatments. PMID- 11902412 TI - Use of antibiotics and roxarsone in broiler chickens in the USA: analysis for the years 1995 to 2000. AB - In 1995, an antibiotic (ANT) was used in starter, grower, and withdrawal (WD) feeds by 94.3, 98.2, and 75.1% of broiler production units, but by 2000, ANT use had declined to 64.8, 66.9, and 48.1% respectively. Roxarsone (ROX) was used in the starter and grower feeds by 69.8 and 73.9% of production units. Bacitracin (BAC) was used more frequently than other antibiotics in the starter and grower feed. Virginiamycin (VIR) was used most frequently in the WD feed. Most units (69.4%) reported use of two different antibiotics. The use of programs comprising two ANT decreased, whereas programs with a single ANT increased during the period of study. A combination of ionophore (ION) + ROX + ANT was employed most frequently in the starter and grower feeds, whereas an ANT alone was used most frequently in the WD ration. The use of ION + ROX + ANT declined from 1995 to 2000, but use of ION + ROX increased. There were no significant differences in calorie conversion whether plants used ION + ROX + ANT, ION + ROX, or ION + ANT. The number of days to rear birds to 2.27 kg was significantly greater for production units using ION + ROX. Mortality was lower for units that used ION + ROX + ANT and ION + ROX than for those that used ION + ANT. Production units that used ION + ANT were more likely to rear birds to a weight greater than 2.5 kg than to 2.0 to 2.5 kg. Units in the South and Central states were more likely to use an ION + ROX than those in the Northeast and Atlantic states, whereas for ION + ROX + ANT the reverse was the case. The cost of medicating with ION + ROX + ANT decreased from 1995 to 1998. PMID- 11902413 TI - Morphological changes in heart and lungs of broilers experiencing pulmonary hypertension syndrome caused by Enterococcus faecalis. AB - In a previous report, a method of identification of birds experiencing early symptoms of pulmonary hypertension syndrome (PHS) caused by challenge with Enterococcus faecalis was delineated. This method involved subjective heart scores based on visual observation of a cavity on the external surface of the right ventricular wall (RVW), as well as tonicity and thickness of this wall. Accuracy in identifying birds 48 h postchallenge with E. faecalis was acceptable. However, this method did not attempt to offer other morphological or physiological characteristics for further understanding the etiology of PHS. In the present study, three trials were conducted to establish morphological characteristics of the heart from birds challenged with E. faecalis. In Trials 1 and 2, discrepancies were found in heart length (HL) and thickness of the RVW. In Trial 3, the dry weight of the right ventricle (RV) increased after challenge with E. faecalis, as was the ratio of the mass of the RV to the mass of the total ventricle (TV). Histopathological evidence of hearts and especially lungs of birds challenged with E. faecalis were suggestive of PHS. Results indicated that RV, RV:TV ratio, and histopatholgical evaluation of heart and lungs are complementary to diagnosis of PHS. PMID- 11902414 TI - Melatonin and the enhancement of immune responses in immature male chickens. AB - Understanding the role of melatonin in affecting different physiological functions, especially immune responses, is becoming increasingly important in the basic and applied sciences. Enhancing the immune response will result in increasing disease resistance and, therefore, improve production efficiency. The purpose of the present study was to investigate the effects of melatonin, administered during the light or dark period, on BW, feed consumption (FC), and immune responses of immature chickens. Eight-week-old Cornell White Leghorn males were used in this study. The doses of melatonin were 0, 5, 10, 20, and 40 mg/kg BW. Melatonin was administered s.c. every 24 h for 7 consecutive d. The chicks were randomly divided into two groups; one group received injection during the middle of the light period, and the other group received injection during the middle of the dark period. All birds received 16 h light and 8 h darkness during a 24-h period. Body weights were measured before and after melatonin treatment, and FC was also measured. After the seven injections, blood samples were collected from the brachial vein, and total white blood cell (WBC) counts, differential cell counts, and activities of T and B lymphocytes were measured. Body weight was not significantly affected by dose of melatonin or time of injection. Furthermore, melatonin did not significantly affect FC; however, FC was significantly lower in the group that was injected in the dark vs. light period. The WBC counts of birds injected with 40 mg melatonin/kg BW were significantly higher than the WBC counts of saline-injected birds. The heterophil/lymphocyte (H/L) ratios of birds injected during the light period were significantly higher than those of birds injected during the dark period. T- and B-lymphocyte proliferation were significantly higher in birds injected with 40 mg melatonin/kg BW compared to saline-injected birds. These results indicate that melatonin in vivo is important in enhancing not only circulating WBC but also activities of B and T lymphocytes of immature male chickens without adversely affecting BW. PMID- 11902415 TI - Effect of vitamin E and A supplementation on egg yolk alpha-tocopherol concentration. AB - The objective of this study was to asses the effect of dietary dl-alpha tocopheryl acetate (alpha-TAC) and vitamin A supplementation on egg yolk alpha tocopherol (alpha-T) concentration. A total of 96 ISA brown hens (a brown-egg laying hen), 32 wk of age, was used. Eight concentrations of alpha-TAC (0, 20, 40, 80, 160, 320, 640, and 1,280 IU/kg diet) were included in a corn-soybean meal based diet containing 4,000 IU of vitamin A/kg diet. Production parameters were not affected by dietary treatment. Interhen variation in yolk alpha-T concentration increased at higher dietary intakes. Logarithmic transformation stabilized variance and showed the existence of a linear relationship between dietary alpha-TAC supplementation level and alpha-T egg yolk concentration (P < 0.001). As the dietary concentration of alpha-TAC rose, efficiency of uptake of alpha-T from feed to yolk decreased linearly (P < 0.001). A parallel experiment was carried out in which four additional groups of hens were fed diets containing four levels of dietary alpha-TAC (0, 40, 160, and 640 IU/kg) and 40,000 IU vitamin A/kg diet. A significantly lower egg yolk alpha-T concentration was found in hens fed diets including the highest level of vitamin A supplementation (P < 0.001). No interaction of dietary alpha-TAC and vitamin A concentrations was observed on egg yolk alpha-T concentration. No effect of dietary treatment was observed on yolk oxidation and other egg quality characteristics. PMID- 11902416 TI - A comparison of eight grades of fat as broiler feed ingredients. AB - Seven samples of feed- or pet food-grade fats (feed- and pet food-grade poultry greases, restaurant grease, white grease, animal/vegetable oil blend, palm oil, yellow grease) and one food-grade edible fat (soybean oil) were evaluated for quality and fatty acid composition. Active oxygen method (AOM) stability at 20 h ranged from 2 to 370 meq/kg; iodine value from 78 to 130 g/100 g; total moisture, insolubles, and unsaponifiables from 0.46 to 3.33%; initial peroxide values from 0.2 to 18.4 meq/kg; and free fatty acids from 0.08 to 21.0%. The ME of the fats ranged from 7.1 to 12.7 kcal/g and was positively correlated with AOM stability and iodine value. When the fats were incorporated into corn-and-soybean-meal based diets at 3 or 6%, no differences in live performance due to fat source were observed. Increasing fat level from 3 to 6% decreased feed conversion by 3.4 points (1.628 vs. 1.662 g/g). Feeding feed-grade poultry grease resulted in significantly smaller abdominal fat pads compared to the other fat sources. Only moisture, insolubles, unsaponifiables, and free fatty acids were significantly correlated with performance responses. Differences were noticed in abdominal fat pad color (lightness and redness) due to fat source. Differences in MEn were not reflected in differences in bird performance. PMID- 11902417 TI - Nutrient use in chicks during the first week posthatch. AB - We examined effects of feeding chicks differing levels of macronutrients on performance during the first 7 d posthatch in this study. Four experiments were conducted using male Ross x Ross broiler chicks. The first experiment examined, in a 3 x 3 factorial design, the effect of feeding fat at 3, 7, and 11% and protein at 18, 23, and 28%. With increasing protein and fat levels feed intake and BW decreased. The second experiment examined, in a 3 x 2 factorial design, the effect of fat at 3, 7, and 10% and cellulose at 3 and 13%. Increasing fat in the diet decreased feed intake at both cellulose levels with little effect on BW. High cellulose decreased feed intake and BW. Experiment 3 examined, in a 3 x 2 factorial design, the effect of protein at 19, 23, and 27% and cellulose at 3 and 13%. Increasing protein levels decreased feed intake with little effect on BW. Cellulose decreased feed intake as levels increased. The fourth experiment examined, in a 4 x 2 factorial, the effect of feeding protein at 19, 21, 23, and 25% and fat at 4.5 and 9%, for 1 wk followed by standard diets through marketing. Significant differences in feed intake, BW and feed efficiency were observed at 7 d, but by 18 d differences were not significant and by 31 and 41 d no treatment effects were observed. Fat and protein percentage in the carcass in all experiments were not altered by dietary treatments and thus the efficiency of protein and fat retention decreased with increasing dietary intake. This study indicates that feeding diets with varying macronutrient levels to chicks during the first week post-hatch has distinct effects as compared to older broilers. It appears that once limiting amino acids and energy are provided the influence of dietary composition on immediate posthatch growth is limited. PMID- 11902418 TI - Effects of age on nutrient digestibility in chicks fed different diets. AB - Three experiments were conducted to determine the effects of age on apparent MEn and apparent amino acid (AA) digestibility of various diets for New Hampshire x Columbian (Experiments 1 and 2) and commercial broiler (Experiment 3) male chicks. Excreta were collected at 0 to 2, 3 to 4, 7, 14, and 21 d of age in all experiments, and nutrient digestibility was determined using acid-insoluble ash as a marker. The first experiment evaluated a corn-soybean meal (SBM) diet. Both MEn and digestibility of AA increased with age, and broken-line regression analysis predicted a plateau at 14 d for MEn and 10 d of age for AA digestibility. For example, MEn increased from 2,970 to 3,430 kcal/kg DM, and lysine digestibility increased from 78 to 89% between 0 and 14 d of age. The second experiment evaluated cornstarch-crystalline AA, dextrose-casein, corn-SBM, and corn-canola meal diets. The MEn of the corn-SBM, corn-canola meal, and cornstarch-crystalline AA diets increased from 0 to 14 d of age. In contrast, the MEn of the dextrose-casein diet was high (3,800 kcal/kg DM) immediately after hatching and did not increase substantially with increasing age. Digestibility of AA increased with age for the corn-SBM and corn-canola meal diets, and broken line regression analysis again predicted a plateau at approximately 10 d of age. Conversely, AA digestibility of the dextrose-casein and cornstarch-crystalline AA diets was high immediately after hatching (93 to 96%) and increased only slightly with age. The results of Experiment 3 with commercial broiler chicks also showed significant (P < 0.05) increases in MEn and AA digestibility with increasing age. The results of this study indicated that the MEn and AA digestibility of corn-SBM and corn-canola meal diets increase with age for young chicks. The results also showed that MEn and AA digestibility were very high for a dextrose-casein diet immediately after hatching. Thus, the latter ingredients may have beneficial effects for very young chicks. PMID- 11902419 TI - Changes in the populations of mitotic and apoptotic cells in white follicles during atresia in hens. AB - The aim of this study was to determine whether the population of mitotic cells changes in correlation with apoptotic cell population in follicular tissues during atresia of white follicles in hens. Hens were injected with 5-bromo-2' deoxyuridine (BrdU) 1 h before tissue collection. The small white follicles were classified as healthy follicles and as early or late atretic follicles by histological observation. Mitotic and apoptotic cells were determined by immunocytochemistry for BrdU and terminal deoxytranceferase-mediated dUTP nick end labeling (TUNEL), respectively. The BrdU labeling was observed in some of the granulosa cells and thecal fibroblast-like cells in healthy follicles, whereas the population of the labeled cells was reduced in the granulosa and theca layers of atretic follicles. The image analysis confirmed that the frequency of BrdU positive cells declined significantly in the granulosa and theca layers of early atretic follicles compared with those of healthy follicles. In contrast, the TUNEL-positive cells were negligible in healthy follicles. However, they were localized in the granulosa and theca layers of early and late atretic follicles, and those in the theca layer were more inside than outside. The frequency of TUNEL-positive cells was significantly increased with the progress of atretic changes. These results suggest that the population of mitotic cells decreases in association with increase of apoptotic cells during the atretic process of white follicles. PMID- 11902420 TI - Characterization of Staphylococcus aureus isolated from poultry meat in Spain. AB - Ninety-six Staphylococcus aureus isolates from retail chicken carcasses in Spain were characterized using cultural and biochemical tests. The strains were phage typed with the international bacteriophage set for typing S. aureus of human origin. Eighty-eight (91.7%) strains were of the poultry ecovar. Strains of human ecovar were not found. These facts are congruent with findings of other authors. Ninety (93.7%) strains were phage typeable. Lysis by phages of Group III was the most frequent with 66 (68.7%) sensitive strains. Twenty-eight (29.2%) strains were sensitive at 100 routine test dilution (RTD) and only 16 (16.7%) at RTD. By using reversed phage typing, we managed to increase the number of phage typeable strains by 46 (47.9%). More than one S. aureus phage type was detected in 14 (35%) carcasses, which emphasizes the convenience of subtyping several S. aureus isolates from the same food sample in epidemiological studies. Two phage patterns (75/84 and 6/1030/ W57) were the most common. The S. aureus isolates were closely related, as 78 strains showed the most common or indistinguishable (<2 phage reaction differences) phage patterns. PMID- 11902421 TI - The relationship between raw broiler breast meat color and composition. AB - Experiments were conducted to compare the chemical composition of broiler breast meat that was naturally lighter than normal, normal, and darker than normal. In each of three separate replicated trials (wk), fillets were obtained from three commercial processing plants. Approximately 25 fillets of each color group were selected based on International Commission on Illumination (CIE) lightness values as follows: lighter than normal (L* > 53), normal (48 < L* < 51), and darker than normal (L* < 46). The fillets from each replicate, plant, and color group were ground and mixed together, and samples for the 27 treatment groups subjected to color, pH, and chemical analyses (protein, ash, moisture, total lipids, iron, glycogen, and fatty acids profile). The whole fillets had significantly different color values for the three color groups at 0 and 24 h prior to grinding. Of the ground meat samples, there were significant treatment and plant differences in composition. There were no color treatment effects on moisture, lipid, glycogen, iron, ash, or fatty acid ratios. Meat from the light group had significantly lower protein values than the normal or dark meat and lower ash than the dark group. The light group also had significantly higher levels of C16:1 and lower levels of C18:0 and C20:4 fatty acids than the dark group. Among the three plants, there were significant effects for breast meat color and composition. Results indicated that, although plant had more effect on composition, differences by color group might indicate that extreme variation in color may be due to long-term genetic factors as well as short-term antemortem stress. PMID- 11902422 TI - Effect of dietary conjugated linoleic acid on the growth rate of live birds and on the abdominal fat content and quality of broiler meat. AB - Two experiments were conducted to investigate the influence of dietary conjugated linoleic acid (CLA) on the growth, body composition, abdominal fat accumulation, and meat quality in broilers. In Experiment 1, 50 broilers that were 3 wk old (total of 200 birds) were assigned to one of the four diets containing 0, 0.25, 0.5, or 1% CLA and were fed for 3 wk. In Experiment 2, 40 broilers that were 3 wk old (total of 120 birds) were assigned to one of the three diets containing 0, 2, or 3% CLA and fed for 5 wk. At the end of the 3-wk feeding trial, the average body weight of broilers for Experiment 1 was about 2.20 kg per bird for all treatments. For Experiment 2, after 5-wk feeding trial, the average body weights of birds were 4.04, 3.99, and 3.93 kg for the control, 2% CLA, and 3% CLA groups, respectively, with a non significant reduction in body weight as the levels of dietary CLA increased. There was no difference in abdominal fat weight, the total body fat, and protein content in broilers among the CLA treatments for Experiment 1. However, when the dietary CLA was increased to 2 or 3%, the total body fat content was reduced. The whole body fat content decreased from 14.2% in the control to 11.9 and 12.2% for 2 and 3% CLA, respectively. Dietary CLA at 2 and 3% levels influenced meat quality. After cooking, the breast meat from 2 or 3% dietary CLA treatment was harder and drier, and the color was a little darker than that of the control. These changes could be caused by the decreased unsaturated fatty acid content in meat after CLA feeding, which increased the melting point of the fat. PMID- 11902423 TI - REM sleep behavior disorder: clinical, developmental, and neuroscience perspectives 16 years after its formal identification in SLEEP. PMID- 11902424 TI - Practice parameters for the use of auto-titrating continuous positive airway pressure devices for titrating pressures and treating adult patients with obstructive sleep apnea syndrome. An American Academy of Sleep Medicine report. AB - Continuous positive airway pressure (CPAP) is used to treat patients with the obstructive sleep apnea syndrome (OSAS). The current standard is for an attendant technician to titrate CPAP during full polysomnography to obtain a fixed single pressure. The patient uses CPAP nightly at this fixed single pressure. Recently, devices using new technology that automatically titrate positive airway pressure (APAP) have become available. Such devices continually adjust pressure, as needed, to maintain airway patency (APAP titration). These adjustments can be made with or without attendant technician intervention. Data obtained during APAP titration can be used to provide a fixed single pressure for subsequent treatment. Alternatively, APAP devices can be used in self-adjusting mode for treatment (APAP treatment). A task force of the Standards of Practice Committee of the American Academy of Sleep Medicine reviewed the available literature. Based on this review, the Standards of Practice Committee developed these practice parameters as a guide to the appropriate use of APAP. Recommendations are as follows: 1) A diagnosis of OSAS must be established by an acceptable method. 2) APAP titration and APAP treatment are not currently recommended for patients with congestive heart failure, significant lung disease (e.g., chronic obstructive pulmonary disease), daytime hypoxemia and respiratory failure from any cause, or prominent nocturnal desaturation other than from OSA (e.g., obesity hypoventilation syndrome). In addition, patients who do not snore (either due to palate surgery or naturally) should not be titrated with an APAP device that relies on vibration or sound in the device's algorithm. 3) APAP devices are not currently recommended for split-night studies since none of the reviewed research studies examined this issue. 4) Certain APAP devices may be used during attended titration to identify by polysomnography a single pressure for use with standard CPAP for treatment of OSA. 5) Once an initial successful attended CPAP or APAP titration has been determined by polysomnography, certain APAP devices may be used in the self-adjusting mode for unattended treatment of patients with OSA. 6) Use of unattended APAP to either initially determine pressures for fixed CPAP or for self-adjusting APAP treatment in CPAP naive patients is not currently established. 7) Patients being treated with fixed CPAP on the basis of APAP titration or being treated with APAP must be followed to determine treatment effectiveness and safety, and 8) a re-evaluation and, if necessary, a standard attended CPAP titration should be performed if symptoms do not resolve or the CPAP or APAP treatment otherwise appears to lack efficacy. PMID- 11902425 TI - The use of auto-titrating continuous positive airway pressure for treatment of adult obstructive sleep apnea. An American Academy of Sleep Medicine review. AB - This paper reviews the efficacy of auto-titrating continuous positive airway pressure (APAP) for treatment of obstructive sleep apnea. It is based on a review of 30 articles published in peer review journals conducted by a task force appointed by the American Academy of Sleep Medicine to develop practice parameters for use of APAP devices for treatment of obstructive sleep apnea (OSA). The data indicate that APAP can be used to treat many patients with OSA (auto-adjusting) or to identify an effective optimal fixed level of continuous positive airway pressure (CPAP) for treatment (auto-titration). Patients with significant congestive heart failure, chronic obstructive pulmonary disease (COPD), or significant amounts of central apnea were excluded from many treatment trials and there is insufficient evidence that APAP can be used to treat these patients. Many clinical trials have been performed in patients already on CPAP or with the initial APAP night in a laboratory setting. At this time only a few studies have evaluated initial titration with APAP in CPAP-naive patients in an unattended setting. Further studies of APAP in this circumstance are needed. No studies have systematically compared the efficacy of one APAP technology with another. Devices using different technology may not give the same results in a given patient. Devices solely dependent on vibration may not work in non-snorers or patient who have undergone upper-airway surgery. High mask or mouth leaks may prevent adequate titration in devices monitoring snoring, flow, or impedance (forced oscillation technique). Review of the raw data to identify periods of high leak was performed in several of the APAP titration studies, to identify a pressure for fixed CPAP treatment or to determine if the titration was adequate. There is conflicting evidence for and against the premise that treatment with APAP increases acceptance and adherence compared to fixed CPAP. In studies demonstrating an increase in adherence with APAP, there was similar improvement in measures of daytime sleepiness as with fixed CPAP treatment. Further studies are needed to determine if APAP can increase acceptance or adherence with positive pressure treatment in patients with OSA. PMID- 11902426 TI - Clomipramine suppresses postnatal REM sleep without increasing wakefulness: implications for the production of depressive behaviors. AB - Clomipramine (CLI), a REM sleep suppressant, alleviates symptoms of depression in adults but produces depressive behaviors if applied neonatally. Both effects of CLI as applied to adults and to neonates have been interpreted as consequences of its involvement in REM sleep deprivation. However, the paradox of these conflicting effects remains to be understood. The current study attempts to find the possible answer by studying the effects of CLI on postnatal sleep. Eight postnatal rats were evaluated polysomnographically for nine days. Four rats were treated with CLI, 40 mg/kg/day for six days, and four rats were treated with equivolume saline during the same period. The results showed that 1) CLI treatment did not reduce the time of phasic muscle activity which appears during slow wave EEG as it did during REM sleep; 2) during treatment, rats treated with CLI had 44.66%-68.62% REM sleep reduction, varied according to age; 3) REM sleep reduction during treatment was generally compensated by non-REM sleep, so that total sleep (and wakefulness) was comparable to that experienced by rats treated with saline; 4) an obvious REM sleep rebound was observed after drug withdrawal at the age of P19. These results suggest that 1) the stage that shows phasic muscle activity simultaneously with a high amplitude EEG is not REM sleep and is likely to be independent from non-REM sleep in terms of the percentile change; 2) REM sleep reduction without a corresponding increase in wakefulness in postnatal rats is likely the mediator of postnatal RSD in the production of adult depression; and 3) the neuronal bases responsible for REM rebound function by the end of the postnatal third week. PMID- 11902427 TI - The siesta and mortality in the elderly: effect of rest without sleep and daytime sleep duration. AB - STUDY OBJECTIVE: To examine the effects of daytime rest without sleep and duration of the siesta on mortality. DESIGN: Longitudinal observation. SETTING AND PARTICIPANTS: Population sample of 442 community-dwelling 70-year-old subjects examined both at home and in a geriatric hospital. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Overall mortality for those who neither rested nor slept was 9.3%, for those who rested without sleep 10.9%, and for those who slept 19%, p<0.02. Males had higher mortality (19%) than females (10%), p<0.006. Rest without sleep had no effect on mortality: in males, 13%, 21%, 18% for no rest, rest of less than one hour, or more than one hour with respective 8%, 6% and 4% in females. However, daytime sleep in males of more than one hour had 28.2% mortality, whereas sleep of one hour or less had 13.6% mortality, p=0.02. In females mortality rate was not different by sleep duration: 16% and 13.6% for those who slept for one hour or less, or more than one hour, respectively. In a multivariate analysis in females, a siesta of one hour or less was associated with risk odds ratio (ROR) of 4.67 and 95% confidence interval (CI) 1.22-17.80 and of one to two hours, ROR was 5.57 and 95% CI 1.05-29.49. For males, siesta of less than one hour was not associated with excess risk, ROR 0.9 and 95% CI 0.39 2.38; a siesta of one to two hours with ROR of 2.61 and 95% CI 1.01-6.80; and two hours or more ROR 13.6 and 95% CI 0.98-2.10. CONCLUSIONS: Rest without sleep is not associated with excess risk of mortality. However, siesta of one to two hours is associated with increased mortality in males whereas, in women, a siesta of less than one hours confers the excess risk. PMID- 11902429 TI - The epidemiology of narcolepsy in Olmsted County, Minnesota: a population-based study. AB - STUDY OBJECTIVES: To determine the age- and sex-specific incidence rates and prevalence of narcolepsy in a United States community. DESIGN: The records linkage system of the Rochester Epidemiology Project was utilized to ascertain all patients with narcolepsy seen in Olmsted County, Minnesota between 1960 and 1989. Age- and sex-specific incidence rates were calculated, using census data. Prevalence of narcolepsy on January 1,1985 was calculated. SETTING: N/A. PATIENTS OR PARTICIPANTS: Community patients diagnosed with narcolepsy by a validated set of diagnostic criteria. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The incidence rate per 100,000 persons per year was 1.37 (1.72 for men and 1.05 for women). The incidence rate was highest in the 2nd decade, followed in descending order by the 3rd, 4th and 1st decades. The prevalence on January 1, 1985 was 56.3 per 100,000 persons. Approximately 36% of prevalence cases did not have cataplexy. CONCLUSIONS: Narcolepsy is not a rare disorder. It appears to be commoner in men. It originates most commonly in the 2nd decade. Narcolepsy without cataplexy is an important subgroup, warranting further study. PMID- 11902428 TI - ICSD diagnostic criteria for narcolepsy: interobserver reliability. Intemational Classification of Sleep Disorders. AB - STUDY OBJECTIVES: To estimate the reliability of the diagnosis of narcolepsy after clinical interview and polysomnographic evaluation among sleep medicine doctors, before and after training in application of the International Classification of Sleep Disorders (ICSD). SETTING: Videotaped semi-structured interviews of 10 patients complaining of daytime sleepiness of different etiologies. Questions referred to ICSD criteria for narcolepsy. A further series of 10 cases of narcolepsy without cataplexy were simulated, with at least a random one to three of the ICSD polysomnographic criteria at pathological levels. PARTICIPANTS AND DESIGN: Seventeen doctors were required to classify each videotaped case as "ascertained," "possible," or "excluded" narcolepsy, in two sessions: one before and one after discussion of ICSD criteria. The observers were invited to confirm or exclude the diagnosis of narcolepsy in the 10 simulated cases, according to the given polysomnographic findings, before and after an agreed proposal of the interpretation of ICSD polysomnographic criteria. Interobserver reliability was calculated using Kappa statistics. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Interobserver reliability of clinical judgement improved from "substantial" at baseline (Kappa 0.61) to "almost perfect" after training (Kappa 0.95). Interobserver reliability of polysomnographic findings was "fair" at baseline (Kappa 0.24), unanimous after the proposed interpretation of ICSD polysomnographic criteria. CONCLUSIONS: Baseline reliability of diagnostic judgement in suspected narcolepsy was found satisfactory among Italian sleep medicine doctors. Educational training, based on discussion of ICSD criteria, further improved agreement. Diagnosis based on polysomnographic findings, not reliable at baseline, needed a strict interpretation of ICSD criteria to attain standardization. PMID- 11902430 TI - Nightmare prevalence, nightmare distress, and self-reported psychological disturbance. AB - STUDY OBJECTIVES: The relationship between nightmare prevalence, nightmare distress, and self-reported psychological disturbance was assessed prospectively. DESIGN: Differences in self-reported psychological disturbance as a function of nightmare prevalence was investigated by MANCOVA's with non-nightmare dreams as the covariate as well as Pearson correlations. The relative contribution of nightmare prevalence and distress to the prediction of psychological disturbance was investigated through multiple regression analyses. SETTING: N/A. PARTICIPANTS: 116 participants (mean age = 20 years) completed self-report indices of depression, anxiety, dissociation, psychosis-proneness, and a psychiatric symptom checklist and kept a nightmare log for 21 consecutive nights. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Frequent nightmares were associated with higher levels of psychological disturbance. Individuals who reported 3 or more nightmares across the 3 weeks reported more dissociation, psychosis proneness and psychiatric symptoms than participants reporting 2 nightmares or less. However, nightmare prevalence and distress were not significantly correlated and differentially predicted to different types of waking psychological disturbance. Multiple regressions further indicated that nightmare distress accounted for much of the unique explanatory variance in predicting clinical states associated with high negative affect (anxiety and depression). Last, there was no evidence for a specific relationship between nightmares and psychosis-proneness. CONCLUSIONS: The findings suggest that it is not the incidence of nightmares which is associated with poorer waking psychological functioning, especially anxiety and depression states, but the reported distress associated with the nightmare experience which is the critical variable in predicting higher psychological disturbance. PMID- 11902431 TI - Associations between symptoms of inattention, hyperactivity, restless legs, and periodic leg movements. AB - STUDY OBJECTIVES: Attention-deficit/hyperactivity disorder (ADHD) has shown associations with restless legs syndrome (RLS) and periodic leg movements during sleep (PLMS) among small samples of referred children, but whether RLS or PLMS are common more generally among hyperactive children has not been well studied. DESIGN: Cross-sectional survey. SETTING: Two university-affiliated but community based general pediatrics clinics. PATIENTS: N=866 children (469 boys), aged 2.0 to 13.9 years (mean 6.8+/-3.2 years), with clinic appointments. INTERVENTIONS: N/A. MEASUREMENTS: A validated Pediatric Sleep Questionnaire assessed for PLMS (a 6-item subscale), restless legs, growing pains, and several potential confounds of an association between behavior and PLMS or RLS. Parents also completed two common behavioral measures, a DSM-IV-derived inattention/hyperactivity scale (IHS) and the hyperactivity index (HI, expressed as a t-score) of the Conners' Parent Rating Scale. RESULTS: Restless legs were reported in 17% (95% C.I. [15, 20]) of the subjects. Positive HI scores (>60) were found in 13% [11, 16] of all subjects, 18% [12, 25] of children with restless legs, and 11% [9, 14] of children without restless legs (chi-square p<0.05). Odds ratios between HI>60 and each of the following were: a one-s.d. increase in the overall PLMS score, 1.6 [1.4, 1.9]; restless legs, 1.9 [1.1, 3.2]; and growing pains, 1.9 [0.9, 3.6] (all age and sex-adjusted). Results were similar for high IHS scores (>1.25). The associations between each behavioral measure and the PLMS score retained significance after statistical adjustment for sleepiness, snoring, restless sleep in general, or stimulant use. CONCLUSIONS: Inattention and hyperactivity among general pediatric patients are associated with symptoms of PLMS and RLS. If either condition contributes to hyperactivity, the magnitude of association suggests an important public health problem. PMID- 11902432 TI - Diagnostic codes associated with hypnotic medications during outpatient physician patient encounters in the United States from 1990-1998. AB - STUDY OBJECTIVES: This investigation tabulates the most common diagnostic codes associated with use of hypnotic medications during physician-patient encounters in the U.S. DESIGN: Estimates were derived from the National Ambulatory Medical Care Survey (NAMCS) from 1990 through 1998. Diagnoses were coded according to the International Classification of Disease-Clinical Modification-9th Edition (ICD-9 CM). SETTING: The NAMCS collects outpatient visit data from nonfederal physicians of all specialties, except anesthesiology, pathology, and radiology. PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Hypnotic medications were more commonly associated with psychiatric codes than with the symptom of insomnia. Primary insomnia did not appear among the codes most commonly associated with a hypnotic prescription. CONCLUSIONS: Hypnotics were more commonly associated with psychiatric codes than with insomnia codes, but it is unknown whether the coding accurately reflects the true diagnoses. Still, since there is minimal data on the efficacy and safety of hypnotics in persons with psychiatric disorders, these findings may signal a critical knowledge gap in the treatment of insomnia. PMID- 11902433 TI - Actigraph placement and sleep estimation in children. AB - STUDY OBJECTIVES: To determine whether actigraph placement affects sleep estimation in children. DESIGN: Descriptive study. SETTING: Naturalistic setting. PARTICIPANTS: Twenty children aged 7-12 years from primary schools. INTERVENTIONS: N/A. MEASUREMENTS: Motor activity was measured from the waist and non-dominant wrist with actigraphs for three consecutive days during a school week. RESULTS: The minute-by-minute agreement of sleep-wake states between the two measurement sites was 92.5%. Wrist- and waist-recorded sleep parameters correlated well and the mean values did not differ. CONCLUSIONS: Although the placement of the actigraph slightly affected the measured activity parameters, its influence on 3-night mean sleep estimates in children was not statistically significant. PMID- 11902434 TI - Nightcap: a reliable system for determining sleep onset latency. AB - Previous studies have demonstrated that the Nightcap home-based sleep-monitoring system can differentiate waking, NREM and REM sleep based on eyelid movements (ELMs) and head movement behavior. The present study aims at determining the reliability of the Nightcap in determining the human sleep onset latency (SOL) as revealed by standard polysomnography (PSG). Four naps were recorded in each of ten normal subjects using both PSG and the Nightcap simultaneously. The Nightcap algorithm scored sleep onset as the first of 4 consecutive 30-sec epochs with less than 5 ELMs. The mean percentage of agreement between the Nightcap and PSG was 93% (k = 0.79), and the average absolute difference was 45 sec (13.3% of SOL(PSG)). SOL(NC) differed by less than 1 min in 85% of onsets. Recordings of EEG activity from 90 sec before and after PSG-identified sleep onsets were subjected to spectral analysis. Changes in spectral power in the theta (4-7 Hz) and alpha (8-12 Hz) frequency bands during the transition into light sleep correlated well with eyelid behavior. However, changes in ELM density predicted sleep onset better than did changes in theta and alpha spectral power. These results suggest that the Nightcap may be a potential alternative to the PSG technique in the assessment of SOL in normal subjects. PMID- 11902435 TI - Chronic behavioral disorders of human REM sleep: a new category of parasomnia. 1986 [classical article]. PMID- 11902436 TI - Marine natural products. AB - This review covers the marine natural products literature for the year 2000 and is organized phylogenetically, with sections on marine microorganisms and phytoplankton, green algae, brown algae, red algae, sponges, coelenterates, bryozoans, molluscs, tunicates. echinoderms and miscellaneous marine organisms. There is an emphasis on new structures, stressing their biological activities, source organisms and countries of origin, and also syntheses that confirm the structures of known compounds. The review contains 869 structures and 592 references, of which 434 appeared between January and December 2000. PMID- 11902437 TI - Glycopeptide antibiotics and development of inhibitors to overcome vancomycin resistance. AB - This review outlines the history of vancomycin and other key glycopeptide antibiotics and the molecular basis of vancomycin resistance, and focuses on the development of synthetic inhibitors of vancomycin-resistant enzymes VanR, S, A, H and X to overcome resistance. The literature up to July 2001 is reviewed and 119 references are cited. PMID- 11902438 TI - The oxylipin chemistry of attraction and defense in brown algae and diatoms. AB - This review covers the research on brown algal pheromones from the first structural characterisation of an active principle in 1971 to the recent detailed insight into their biosynthesis. Development of analytical methods and bioassays that lead to the identification of a structural variety of different fatty acid derived pheromones are reported. Special emphasis is focused on the inactivation of initially released pheromones through pericyclic reactions. The impact of pheromone-research on the defensive chemistry of brown algae and diatoms is discussed. 121 references are cited. PMID- 11902439 TI - Insights on the conformational stability of collagen. AB - This review describes work on the conformational stability of the collagen triple helix. In 1994, the structure of collagen was determined at high resolution. Since then, much work has been done on synthetic mimics of collagen that contain host-guest peptides, tethers, peptoid residues, or analogs of the prevalent 4(R) hydroxy-L-proline residues. This work has revealed much about the chemical basis for collagen stability, and could spawn useful new biomaterials. The literature from 1994 to mid 2001 is reviewed, and 116 references are cited. PMID- 11902440 TI - From Lobry de Bruyn to enzyme-catalyzed ammonia channelling: molecular studies of D-glucosamine-6P synthase. AB - This review summarizes the state of knowledge on D-glucosamine-6P synthesis catalyzed by glucosamine-6P synthase. The mechanisms of L-glutamine hydrolysis, ammonia transfer and fructose-6P conversion into D-glucosamine-6P are analyzed with the E. coli enzyme in light of recent X-ray structures. With 92 references this paper covers the literature up to June 2001 and emphasizes the potential implication of the mammalian glucosamine-6P synthase in type 2 diabetes. PMID- 11902441 TI - Biosynthesis and attachment of novel bacterial polyketide synthase starter units. AB - The biosynthesis and mode of attachment of a wide range of polyketide synthase (PKS) starter units in bacteria are covered in this review. Natural, unnatural, and engineered starter units associated with type I and type II PKSs are reported. The literature through early 2001 is reviewed, and 240 references cited. PMID- 11902442 TI - Renal cell neoplasia. PMID- 11902443 TI - Light microscopic and electron microscopic diagnosis of gastrointestinal opportunistic infections in HIV-positive patients. AB - The surgical pathologist is expected to recognise gastrointestinal opportunistic infections in biopsies from HIV-positive patients, and patients immunocompromised iatrogenically by cancer therapy, steroid treatment and transplantation immunosuppression regimes. This review article presents the diagnostic features in gastrointestinal biopsies of microsporidia, cyclospora, isospora, cryptosporidia, mycobacteria, adenovirus, enteropathogenic bacteria, cryptococcus and leishmania. All of these infections have been diagnosed at our hospital in Sydney, Australia, since the AIDS epidemic began. A protocol for the examination and assessment of these gastrointestinal biopsies is presented and discussed. PMID- 11902444 TI - Cutaneous lymphomas: which pathological classification? AB - Cutaneous lymphomas are rare and although some are a manifestation of systemic lymphoma, the majority arise primarily from the skin. These primary cutaneous lymphomas comprise predominantly T cell subtypes and represent a wide spectrum of disorders. Pathologists can currently choose to label these conditions according to three classifications (REAL, EORTC or WHO) but each has shortcomings. Nonetheless, in an attempt to unify the field, we would recommend that pathologists make every attempt to categorise these conditions according to the WHO classification. This classification can encompass all the conditions and aligns the cutaneous lymphomas with the broader systemic lymphoproliferative conditions. PMID- 11902445 TI - Identification of tumours with the CD43 only phenotype during the investigation of suspected lymphoma: a heterogeneous group not necessarily of T cell origin. AB - AIMS: CD43 is usually employed as a T cell marker in the immunophenotypic work-up of suspected cases of non-Hodgkin's lymphoma (NHL). In this setting, tumours expressing CD43 in the absence of other T or B cell markers (CD43 only phenotype) are rare. We present four cases with this aberrant phenotype seen at our institution. METHODS: The CD43 only phenotype was defined as expression of CD43 in the absence of expression of B cell markers CD20 and CD79a, and T cell markers CD3 and CD5, on initial immunohistochemistry performed on biopsies of suspected NHL. Combinations of further immunohistochemistry, flow cytometry, cytogenetic analysis and molecular studies were used to enable further diagnosis and lineage assignment. RESULTS: The four cases were subsequently diagnosed as: one case of extramedullary acute myeloid leukaemia, one case of null cell anaplastic large cell lymphoma, and two cases of extranodal diffuse large B cell lymphoma. None were demonstrated to be of T cell origin. CONCLUSIONS: Our series further confirms the lack of specificity of CD43 expression for T cell lineage. Documentation of the CD43 only phenotype in suspected cases of NHL therefore requires further investigation to both correctly diagnose and clarify lineage of these tumours. PMID- 11902446 TI - Intestinal epithelial lesions associated with signet ring cell carcinoma of the colon and small intestine. AB - AIMS: The purpose of this study was to investigate the epithelial lesions associated with signet ring cell carcinoma (SRCC) of the colon and small intestine and the possible mechanism of tumour development. METHODS: Twenty-seven cases of adenocarcinoma with a signet ring cell (SRC) component of the colon and small intestine were divided into three groups depending on the association of the SRCC with: (1) epithelium without definite epithelial dysplasia, (2) adenoma, and (3) common type of adenocarcinoma (CTCA) with SRCC component occupying more than 50%, 30%, or less than 30% of the tumour. RESULTS: Most carcinomas were of T3 or T4 type, using the TNM standard staging system. The SRCC component was histopathologically similar in all groups. In group 1 (four cases, linitis plastica type), the overlying epithelium was normal or showed indefinite epithelial dysplasia and occasionally contained intra-epithelial SRCs. In groups 2 and 3 (two and 21 cases, respectively), seven cases contained multiple foci of intra-epithelial SRCs in areas separated from the invasive carcinoma. Transitional areas between SRCC and adenoma or CTCA were also identified. Immunostaining for p53 showed a varied extent of positive reactivity in 23 SRCC. The degree and the extent of reactivity appeared to increase with the stage of the carcinoma. Most intra-epithelial SRCs were immunoreactive for p53. Linitis plastica SRCC was associated with extensive p53 reactivity of the 'atypical' and the adjacent 'normal' epithelium. CONCLUSIONS: SRCC may arise from either CTCA, adenoma, 'atypical' epithelium or a combination of these epithelia. SRCC accounts for the bulk of carcinoma in each of these categories. In linitis plastica SRCC, positive reactivity for p53 is extensive in the adjacent 'normal' colonic epithelium and extends as far as 3cm from the microscopically identified SRCC margin. PMID- 11902447 TI - Double pituitary adenomas: a series of three patients. AB - Multiple pituitary adenomas may occur in up to 1.6-3.3% of patients with Cushing's syndrome. We report three patients with double pituitary adenomas detected at surgery. Two patients had Cushing's disease, but trans-sphenoidal exploration revealed a small prolactinoma in each. One prolactinoma also contained small numbers of basophils. Re-operation in both patients because of persistent Cushing's syndrome showed an ACTH-secreting micro-adenoma. The third patient with acromegaly had two macro-adenomas discovered in different parts of the gland at surgery: one plurihormonal and one null cell tumour. Careful evaluation of pre-operative MRI may not always detect more than one pituitary adenoma. PMID- 11902448 TI - Spitz naevus versus Spitzoid melanoma: when and how can they be distinguished? AB - Spitz naevus is a benign melanocytic lesion that shares many histological features with melanoma. While Spitz naevi characteristically occur in children and young adults and melanomas in the middle-aged and elderly, either tumour can occur in patients of any age. In many cases, the histopathological diagnosis of Spitz naevus is straightforward, particularly in small lesions displaying many or all of the typical histological features and occurring in young patients. Tumours that deviate from the classic description, however, cause difficulties in diagnosis. In this review, we highlight histopathological features of Spitz naevi and those that may be useful in distinguishing Spitz naevi from melanomas. We find that the presence of good symmetry, Kamino bodies, and uniformity of cell nests or sheets from side-to-side favours a Spitz naevus. The presence of abnormal mitoses, a dermal mitotic rate of >2/mm2, and mitotic figures within 0.25 mm of the deep border of the lesion favours a melanoma. Immunohistochemical stains for HMB45 and Ki67 sometimes provide additional useful information. Despite this, in some cases it may not be possible to give an unequivocal diagnosis. Recommendations for the reporting of such cases are provided. New techniques have also demonstrated chromosomal, molecular and genetic differences between Spitz naevi and melanomas. This report highlights these new data and speculates on their possible future role in the diagnosis of borderline lesions. PMID- 11902449 TI - Digital image capture applied to electron microscopic examination of renal biopsies. AB - AIMS: Digital image capture systems to replace traditional film cameras are now available for most electron microscopes. For a diagnostic electron microscope laboratory the test of this new technology is in its application to the examination of renal biopsy specimens. METHODS: A long-term comparison is made between the work procedures employed with conventional film photography versus digital image capture in routine renal biopsy examination. RESULTS: Digital image capture has lead to a reduction in turnaround time and allows for more images to be collected per case, providing more diagnostic information. Ultrastructural measurement is made easier, accuracy of patient records is improved and electronic communication of results is more accessible. Significant operational cost savings are also possible. CONCLUSION: A quicker and more comprehensive assessment of renal biopsy specimens is possible using digital image capture for ultrastructural examination. PMID- 11902450 TI - The autopsy: legal and ethical principles. AB - Recent controversies both in Australia and overseas have focused public attention on autopsy practice. It has become apparent that there is an increasing expectation amongst the public for more detailed information concerning autopsy procedures and, in particular, tissue retention. This article attempts to review the legal and ethical issues with respect to postmortem examination in Australia and to contemplate potential offences and penalties should such obligations be breached. PMID- 11902451 TI - Increasing resistance of Helicobacter pylori to clarithromycin: is the horse bolting? AB - AIMS: To determine whether there has been a change in the patterns of susceptibility to various antibiotics of our isolates of Helicobacter pylori over a 5-year period from 1996 to 2000. METHODS: Five hundred and fourteen isolates of H. pylori grown from gastric biopsies were tested for susceptibility to amoxycillin, clarithromycin, metronidazole and tetracycline. The usage of macrolide antibiotics in Australia was examined by calculating the numbers of prescriptions issued under the Australian pharmaceutical benefits scheme between 1992 and 2000. RESULTS: There were no changes in susceptibility of H. pylori to amoxycillin and tetracycline and there was a slight decline in resistance to metronidazole. In contrast, there was a stepwise 4-fold increase from 3.8 to 15.7% in the number of isolates resistant to clarithromycin and a similar increase in the mean minimum inhibitory concentration of clarithromycin during the 5-year period of observation. There was no change in overall macrolide consumption in Australia over this and the preceding 3 years. However, the pattern changed, with erythromycin usage being halved and being replaced by roxithromycin and clarithromycin. CONCLUSIONS: Resistance of H. pylori to clarithromycin is increasing, possibly as a consequence of increased usage of roxithromycin and clarithromycin. More patients are likely to fail to respond to empirical therapy and will need microbiological investigation. PMID- 11902452 TI - Test and teach. A large tumour arising from the cribriform plate. Intracranial schwannoma unrelated to a major cranial nerve. PMID- 11902453 TI - Test and teach. An unusual tumour presenting in the lungs. Metastatic adult granulosa cell tumour of the ovary, microfollicular pattterns. PMID- 11902454 TI - Intravascular lymphomatosis of the lung and liver following eyelid lymphoma in a Chinese man and review of primary pulmonary intravascular lymphomatosis. AB - Intravascular lymphomatosis (IVL) is characterised by an almost exclusive intravascular proliferation of malignant lymphoid cells, with the diagnosis often made only when the illness is in its terminal phase or at autopsy. We detail a case of IVL affecting the lung and liver of a 49-year-old Chinese man presenting primarily with lung symptoms and incidental findings of abnormal serum transaminase levels, the ante-mortem diagnosis being established on transbronchial lung biopsy and percutaneous liver biopsy specimens, respectively. Histology disclosed CD20 + CD5 - CD10 [corrected] - malignant large mononuclear B cells within the lumina of the blood vessels of the affected organs as well as sinusoids of the liver. Significantly, the patient had a history of large B cell lymphoma affecting the eyelid 18 months prior to the angiotropic disease, suggesting a possible link between the more common types of non-Hodgkin's lymphoma and IVL. A brief review of all cases of primary pulmonary intravascular lymphomatosis is also presented. PMID- 11902455 TI - Adult xanthogranuloma of the vulva: case report and review. AB - We report a case of adult xanthogranuloma arising on the labia majora of a 48 year-old female. Although histologically similar to juvenile xanthogranuloma, the clinical presentation and natural history are unique. To our knowledge this is the first case described of this rare histiocytic lesion arising in the vulva. PMID- 11902456 TI - Meningoencephalomyelitis with vasculitis due to varicella zoster virus: a case report and review of the literature. AB - Varicella zoster virus (VZV) encephalitis is associated with large or small vessel vasculopathy. We report the case of a 67-year-old woman with a history of non-Hodgkin's lymphoma and cancers of the breast and colon, who presented with a zosteriform rash and Brown-Sequard syndrome. Despite 10 days therapy with intravenous acyclovir, meningoencephalitis developed and the patient died 15 days after onset of neurological symptoms. Autopsy showed meningoencephalomyelitis with necrotising vasculitis of leptomeningeal vessels, which is a rare complication of VZV, and we review the literature of the nine similar published cases. Polymerase chain reaction of cerebrospinal fluid for VZV was negative 6 days after onset of neurological symptoms, but became positive by day 10. Only one multinucleated giant cell with intranuclear Cowdry type A inclusions was seen within an endothelial cell in a leptomeningeal vessel involved by vasculitis. PMID- 11902457 TI - Multistep carcinogenesis in anaplastic thyroid carcinoma: a case report. AB - We previously established an anaplastic thyroid carcinoma cell line (KOA2) that had double mutations: an N-ras mutation and a p53 gene mutation. To clarify multistep carcinogenesis, we analysed surgical material from the patient from whom KOA2 was derived for abnormalities in the N-ras and p53 genes. The resected material had two histologically different lesions: a follicular neoplasm and an anaplastic carcinoma. The N-ras mutation was observed in both lesions, but the p53 gene mutation only in the anaplastic lesion. These facts indicate that an N ras mutation may induce follicular neoplasm and a subsequent p53 mutation may have caused the follicular neoplasm to transform to anaplastic carcinoma in this patient. This report suggests direct evidence for multistep carcinogenesis in anaplastic thyroid carcinoma. PMID- 11902458 TI - Integrated urban drainage, status and perspectives. AB - This paper summarises the status of urban storm drainage as an integrated professional discipline, including the management-policy interface, by which the goals of society are implemented. The paper assesses the development of the discipline since the INTERURBA conference in 1992 and includes aspects of the papers presented at the INTERURBA-II conference in 2001 and the discussions during the conference. Tools for integrated analysis have been developed, but there is less implementation than could be expected. That is due to lack of adequate knowledge about important mechanisms, coupled with a significant conservatism in the business. However, significant integrated analyses have been reported. Most of them deal with the sewer system and the treatment plant, while few incorporate the receiving water as anything but the object of the loads to be minimised by engineering measures up-stream. Important measures are local infiltration, source control, storage basins, local treatment and real time control. New paradigms have been introduced: risk of pollution due to system failure, technology for water reuse, sustainability, new architecture and greener up-stream solutions as opposed to down-stream concrete solutions. The challenge is to combine the inherited approaches with the new approaches by flexibility and adaptability. PMID- 11902459 TI - Online-simulation of the WWTP to minimise the total emission of WWTP and sewer system. AB - In this paper, the application of a WWTP-Online-Simulation with the objective to reduce the total emission into the receiving waters is explained. Apart from an introduction and a short description of the possible reduction potentials, first results of the current research project (financed by the German ministry BMBF) are presented. Results of the pilot plant with different experiments of increased stormwater inflow than usual and different control strategies showed the possibility to treat stormwater up to the quadruple dry-weather flow while still meeting the effluent values. However, this is not always guaranteed, and thus a monitoring system with integrated control strategies which is adapted to the load case "stormwater" with prognosis load cases becomes necessary. In the presented example, the simulation (Activated Sludge Model 2d) achieved an excellent match with the measured effluent values of the aeration tank (NH4-N, NO3-N) over a period of several months. The most important prerequisites for good (online-) simulation results are the exact knowledge of the plant and the plausibility and alternative concepts for the measured values in case of sensor failure. PMID- 11902460 TI - Real time control of the integrated urban wastewater system using simultaneously simulating surrogate models. AB - The urban wastewater system (sewer and treatment plant) has a major impact on the river water quality of urban streams. To minimise this impact, real time control is a valuable option. Since the ultimate goal of any control strategy is to optimise the quality of the river system, it is useful to take pollutant immissions into account when determining the control strategy and/or the setpoints of the controller. However, a simultaneously simulating model of the complete system is needed in order to allow design and evaluation of such control strategies. In this work an integrated model of the urban wastewater system is created. This has been accomplished by implementing surrogate models of the three subsystems within a single software platform. The coupled submodels are subsequently used in a semi-hypothetical case study to optimise the resulting river water quality. An ammonia sensor in the river has been used to control the amount of water treated biologically in the treatment plant. It was shown that this integrated control could lower the peak ammonia concentration in the part of the river downstream of the treatment plant. Hence, a proof of principle has been given that the use of measurements in the river to perform control actions in the sewer system and the treatment plant is a promising option. PMID- 11902461 TI - The sewer as a bioreactor--a dry weather approach. AB - The sewer is a reactor for chemical and microbial transformations of wastewater. These in-sewer processes affect the quality of the wastewater and thereby the sewer itself, the subsequent treatment and the receiving water quality. The paper focuses on the interactions between the dry weather in-sewer chemical and microbial transformations of the wastewater and the corresponding processes in a downstream located treatment plant. A conceptual understanding of the sewer processes is crucial in this respect. PMID- 11902462 TI - Probabilistic emission and immission modelling: case-study of the combined sewer WWTP-receiving water system at Dessel (Belgium). AB - The impact of the combined urban drainage and WWTP system of the village of Dessel (Belgium) on the Witte Nete receiving water is modelled both in terms of emissions and immissions. The hydrodynamic and water quality modelling is performed both in a deterministic and probabilistic way. For the deterministic modelling, detailed physically based and simplified conceptual models are used in a complementary way. In the probabilistic modelling, the different uncertainties in the deterministic model are classified in input uncertainties, parameter uncertainties and model-structure uncertainties. The probabilistic simulation results can be used in risk analysis and management, for the determination of the major uncertainty-sources and priorities in model improvement, for model bias elimination and for efficient model calibration. PMID- 11902463 TI - A practical application of integrated urban pollution modelling in Flanders (Belgium): the catchment of Tielt. AB - In view of a European Innovation Project (DG XIII) on the application of the Urban Pollution Management (UPM) procedure, a European consortium was set up to carry out pilot studies in Belgium, France, Ireland, Italy and Portugal. On the Belgian pilot catchment of Tielt the impact from sewer system and sewage treatment plant (STP) on the receiving water courses was studied for both the existing situation and for a number of possible improvement schemes, using the Intermittent Standards, described in the UPM procedure. The integrated modelling revealed some interesting conclusions on the relative impact of the schemes considered. PMID- 11902464 TI - Wet weather water quality modelling of a Portuguese urban catchment: difficulties and benefits. AB - This paper discusses the use of water quality deterministic modelling together with an integrated approach to assess the impact of urban stormwater discharges into ephemeral watercourses, based on the study of a Portuguese catchment. The description of the main aspects, difficulties and benefits found during data collection and model calibration and verification is presented, and the associated uncertainties and errors discussed. Experimental results showed a strong short- and long-term impact of sewer discharges on rivers, and confirmed deposition, resuspension and transport of pollutants as important processes for the water quality. However, the resuspension of riverbed sediment pollutants during storms was probably more significant than the direct impact of the urban discharges. The HydroWorks model was used since it allows for the calculation of pollutant build-up on catchment surfaces and in gully pots, their wash-off, and the deposition and erosion of sediments in sewers. However, it uses several constants, which could not be independently calibrated, increasing the uncertainty already associated with the data. River flows have quite different magnitude from the sewer system overflows, which, together with the difficulties in evaluating river flow rates, makes the integrated modelling approach rather complex and costly. PMID- 11902465 TI - Criteria for assessment of the operational potential of the urban wastewater system. AB - Application of real-time control (RTC) is one possible measure to increase the performance of the urban wastewater system. However, the potential and the benefits of control depend strongly on the characteristics of the individual site under question. Conventionally, to evaluate this potential, a detailed feasibility study had to be carried out. In some cases, such a study may well conclude that, for the given site, real-time control does not have any significant potential, thus resulting in unnecessarily having spent precious resources for a detailed study. It would be desirable to have a methodology that allows simple, and cost-effective, screening of sites for which the analysis of real-time control may be beneficial. Earlier research led to the provision of an easy-to-apply scoring system which allows a quick assessment of the RTC potential of controlling flow in sewer systems. However, since this procedure does not take into account water quality aspects, or the treatment plant or the receiving water body, it cannot be used for assessing the potential of RTC of the complete system, let alone for integrated RTC. This paper describes the first part of an on-going project which aims at establishing an enhanced procedure for assessing the real-time control potential for the entire urban wastewater system. After providing a definition of the term "RTC potential", a large number of (partly hypothetical) case studies (varying a number of key parameters of the wastewater system) is simulated, using the simulation tool SYNOPSIS. For each of these sites, a number of real-time control algorithms are developed and optimised, following a general procedure, which allows for local, global and integrated scenarios to be considered. Analysis of the results reveals those system parameters which are of particular significance to the RTC potential of urban wastewater systems. These are discussed and assessed in this paper. Furthermore, the results of a simulation study are provided which indicate a clear potential of integrated control even for many case studies for which local control provides hardly any benefits. Subsequent studies will complement the simulation study by comparison with a number of real case studies in various countries. PMID- 11902467 TI - Sustainable rainwater management in the Emscher river catchment area. AB - The wastewater management system of the Emscher region is currently being radically restructured. The receiving waters currently surviving as open sewers are to be freed of their wastewater burden and reconstituted to a state as natural as possible, while the wastewater is to be routed underground to the treatment plants. Great importance is attached to the most natural possible rainwater management, in order to buffer extreme run-off situations in the watercourses and to minimize the costs for residential-area water management engineering. Rethinking, which in many cases percolates through only slowly, is necessary in many respects for this purpose. A contest has been set up in the Emscher catchment area in order to accelerate this in the existing residential areas. Seepage, decentralized retention, disconnection and discharge into bodies of water and watercourses have been financially supported. The results are presented and the further procedure deriving from them discussed. PMID- 11902466 TI - Options for alternative types of sewerage and treatment systems directed to improvement of the overall performance. AB - Technology for future houses may well include a high-tech water recycling unit that makes tapwater while people drink bottled water of high quality. There may be toilets that produce just a bag of dry fertiliser per year, hopefully without fossil energy. Rainwater infiltration is increasingly replacing storm sewers anyway. Many urban areas of the future could simply be without sewerage systems. Technical feasibility is given even today and economic feasability is coming closer by advances in membrane technology. However, there are more likely scenarios than this. One person produces about 500 litres of urine and 50 litres of faeces per year (= blackwater). The same person, produces in a range of 20,000 to over 100,000 litres of wastewater. Black- and greywater (wastewater without toilet) do have very different characteristics. If blackwater is collected separately with low dilution it can be converted to safe natural fertiliser, replacing synthetic products and preventing spreadout of pathogens and other pollutants to receiving waters. New sanitation concepts are now built in several countries as pilot projects. One example is a vacuum-biogas system for around 400 inhabitants that has been built in Lubeck, Germany. It does perform recovery of resources and energy in an urban area. This type of sanitation can serve around up to 10,000 people and can be arranged in independent modules for larger settlements. Another pilot project based on urine-sorting flush toilets (no-mix toilets) has been built in the rural water-mill museum "Lambertsmuhle" near Cologne, Germany. Urine or yellow water is collected with low dilution and can be used as fertiliser as projects in Sweden have shown--the nutrient composition suits many types of soil. Brownwater (the solids and flush from the sorting toilet) is converted to small volume by a two-chamber composting tank with a filtration system. The compost can be used as soil conditioner. These and other concepts can be economic and show new ways for the many water scarce areas around the world, too. PMID- 11902468 TI - The influence of stormwater treatment on the hydraulic and pollution load- balance for an entire river basin. AB - The installation of about 500 stormwater detention facilities (SDFs) led to a significant drop of the pollution originating from stormwater runoff in the river basin of the Ruhr which covers 4,488 km2. The German technical directives on the design of SDFs are briefly outlined and the specific costs for such plants are given. The average costs for one kilogram COD held back by SDFs in combined systems amount to Euro 3.73 (calculated without consideration of the subsequent removal in the municipal wastewater treatment plant (WWTP)). The tank volume for stormwater storage can be minimised by application of real-time management systems which allow a dynamic operation of all SDFs in a catchment area. PMID- 11902469 TI - Bayesian decision analysis as a tool for defining monitoring needs in the field of effects of CSOs on receiving waters. AB - In recent years, decision analysis has become an important technique in many disciplines. It provides a methodology for rational decision-making allowing for uncertainties in the outcome of several possible actions to be undertaken. An example in urban drainage is the situation in which an engineer has to decide upon a major reconstruction of a system in order to prevent pollution of receiving waters due to CSOs. This paper describes the possibilities of Bayesian decision-making in urban drainage. In particular, the utility of monitoring prior to deciding on the reconstruction of a sewer system to reduce CSO emissions is studied. Our concern is with deciding whether a price should be paid for new information and which source of information is the best choice given the expected uncertainties in the outcome. The influence of specific uncertainties (sewer system data and model parameters) on the probability of CSO volumes is shown to be significant. Using Bayes' rule, to combine prior impressions with new observations, reduces the risks linked with the planning of sewer system reconstructions. PMID- 11902470 TI - Cost benefit risk--a concept for management of integrated urban wastewater systems? AB - Urban wastewater systems should be evaluated and analysed from an integrated point of view, taking all parts of the system, that is sewer system, wastewater treatment plant and receiving waters into consideration. Risk and parameter uncertainties are aspects that hardly ever have been addressed in the evaluation and design of urban wastewater systems. In this paper we present and discuss a probabilistic approach for evaluation of the performance of urban wastewater systems. Risk analysis together with the traditional cost-benefit analysis is a special variant of multi-criteria analysis that seeks to find the most feasible improvement alternative for an urban wastewater system. The most feasible alternative in this context is the alternative that has the best performance, meaning that the alternative has the lowest sum of costs, benefits and risks. The sum is expressed as the Net Present Cost (NPC). To use NPC as a decision variable has the problematic effect, that two alternatives performing completely differently when focusing on environmental cost can have the same NPC. The extreme example is one alternative with high risk and low cost and another with low risk and high cost. In this example it is up to the decision-maker to decide whether she wants to spend the budget on preventive installations or cleaning up after failures in the environment. PMID- 11902471 TI - Sewer losses and interactions with groundwater quality. AB - Inflow/infiltration (I/I) and infiltration/exfiltration (I/E) are interactive processes which dynamically affect sewer and groundwater performance. The incidence and condition of "critical" sewers in the UK are identified together with chemical and bacterial methods of quantifying I/E and its potential impact on sewer performance and on urban groundwater pollution. Whilst the impacts of I/E do not appear to be substantial on the basis of existing evidence, some caution is advocated in respect of long term sewer sustainability. PMID- 11902472 TI - Effective environmental regulation to maximise the benefits of integrated wastewater management. AB - On the 25 November 1999, the Secretary of State for the Environment in England and Wales announced the outcome of the water companies' third Periodic Review. As a result, a major environmental investment programme will be completed during the period 2000/2005. Overall in England and Wales, it is planned that almost 3,600 km of rivers will be protected or improved and 3,800 unsatisfactory CSOs will be improved to allow receiving water objectives to be met. This means that over the next 5-year period an average of nearly 18 discharges will be improved every week. The implementation of the programme will involve a large workload for the Environment Agency in planning and authorising the individual schemes for delivery by the water companies. The paper illustrates how the Urban Pollution Management procedure and associated environmental standards are to be used to underpin the Environment Agency's policy, to encourage the use of integrated wastewater planning and, as a result, ensure that the anticipated environmental improvements will be delivered. PMID- 11902473 TI - Pollution based real time control of wastewater systems. AB - Wastewater systems are traditionally built as static systems to handle a design load. The real load varies, though, and hardly ever equals the design load. This implies that wastewater systems hardly ever operate in an optimum way, especially during wet weather. Real time control (RTC) of regulators can improve the operation by better fit of the system to the actual state and load. RTC based on pollutant concentrations together with hydraulic conditions (pollution based real time control, PBRTC) is investigated in this paper to assess the potential pollutant load reduction on receiving waters at wet weather without expansion of transport or storage capacity. Both CSOs and WWTP effluents contribute to the pollutant discharges to receiving waters and both are considered. Three cases are studied to assess the potential benefit of PBRTC. Giving priority to the most polluted wastewater for treatment and storage in branched interceptor systems can reduce CSO discharge loads by more than 20%. Biological WWTPs and especially activated sludge plants are more complex and less stable than chemical precipitation plants during and after high pollutant and hydraulic load. Biological plants can hence profit more from PBRTC than chemical precipitation plants. Receiving waters that are sensitive to acute effects caused by intermittent discharges can benefit more from PBRTC than receiving waters with problems connected to long-term accumulation of pollution. PMID- 11902474 TI - Effects of real time control of sewer systems on treatment plant performance and receiving water quality. AB - Four individual mathematical submodels simulating different subsystems of urban drainage were intercoupled to an integral model. The submodels (for surface runoff, flow in sewer system, wastewater treatment plant and receiving water) were calibrated on the basis of field data measured in an existing urban catchment investigation. Three different strategies for controlling the discharge in the sewer network were defined and implemented in the integral model. The impact of these control measures was quantified by representative immission state parameters of the receiving water. The results reveal that the effect of a control measure may be ambivalent, depending on the referred component of a complex drainage system. Furthermore, it is demonstrated that the drainage system in the catchment investigation can be considerably optimised towards environmental protection and operation efficiency if an appropriate real time control on the integral scale is applied. PMID- 11902475 TI - The effect of extended in-sewer storage on wastewater treatment plant performance. AB - A project funded by UKWIR is under way in the UK to develop a relatively simple methodology whereby the effects of the introduction of extended in-sewer storage at CSOs on downstream sewerage and treatment can be assessed. Recent legislation (UK and European) has compelled many sewer system operators to introduce systems which increase in-sewer retention times, and also retain more flow and load within sewer networks. The project has reviewed existing knowledge about the interaction between in-sewer flow and treatment plants, together with available models. The study is utilising a "benchmark" of 3 configurations of treatment plant and dynamic simulation using the WRc STOAT software, with minor modifications to ensure that effects on odour generation and nutrient removal processes are adequately modelled. As no existing sewer flow quality model can represent the range of conditions possible in sewer networks, a combined application of the Hydroworks model and a new model developed at Aalborg University is being used for this part of the study. PMID- 11902476 TI - The impact of the controlled emptying of in-sewer storage on wastewater treatment plant performance. AB - The use of in-sewer storage is generally considered to be an effective means of minimising the effects of intermittent discharges into receiving watercourses during combined flows. Despite this, very little information is available about the consequential effects these flows may have on recipient wastewater treatment plant performance. Typical problems may include biomass washout (hydraulic), and reduced biological reactor performance due to dilute loading (biological). A study is described where detailed analysis was carried out to ascertain the consequential effects of prolonged dilute loading on an activated sludge wastewater treatment plant in Perth, Scotland. Consideration was given to likely storage volumes which may have been utilised in the catchment to resolve local problems. A comprehensive analysis of resulting treatment plant performance was carried out for variations in flow and various wet weather loadings. It is concluded that storage may cause little or no benefit with respect to ammonia total emissions due to reduced treatment of dry weather flows subsequent to the prolonged combined loading period. This was exacerbated by the long regeneration times of nitrifying bacteria. However, an overall benefit with respect to BOD total emissions would always be expected, as appropriately sized storage would retain the first foul flush at the CSOs, thereby compensating the increased emissions from the downstream wastewater treatment plant. PMID- 11902477 TI - Biodegradability of wastewater--a method for COD-fractionation. AB - Characterization of wastewater for simulation of in-sewer transformations can be carried out by interpretation of oxygen uptake rate measurements in combination with a conceptual model of the microbial transformations involved. This interpretation can be done by iterative procedures by solving the differential equations constituting the model or by the application of a more "manual" method- the latter being the topic of this paper. Examples where different wastewaters are characterized illustrate the method. PMID- 11902478 TI - Assessment of stormwater impacts on an urban stream with a detention pond. AB - Impacts of urban development on a small creek with an on-stream stormwater pond, which was built to mitigate the effects of a 13-ha commercial plaza on creek flows, were assessed by investigations of the creek-pond system hydrology, water and sediment chemistry and toxicity, and benthic communities. The hydrology and conventional pollutant loadings of this system were dominated by the creek catchment (4.5 km2), which contributed more than 95% of the total flow; the rest was contributed by the plaza runoff which was less polluted than typical residential runoff. Conventional bioassays (Daphnia magna, Microtox) did not indicate any confirmed acute toxicity in the creek flow, plaza drainage, or pond outflow. However, sediments accumulated in the pond were rather polluted and several sediment bioassays (including Sediment Microtox) indicated their severe toxicity. In benthic community assessments, taxa richness and total counts of benthic organisms did not change much when moving from upstream to downstream of the pond. Thus, the pond accumulates sediments and toxicants and thereby prevents further degradation of the creek condition downstream of the plaza drainage outfall. PMID- 11902479 TI - Managing sewer solids for the reduction of foul flush effects--Forfar WTP. AB - In times of high sewer flow, conditions can exist which enable previously deposited material to be re-entrained back into the body of the flow column. Pulses of this highly polluted flow have been recorded in many instances at the recently constructed wastewater treatment plant (WTP) in Forfar, Scotland. Investigations have been undertaken to characterise the incoming flows and to suggest remedial measures to manage the quality fluctuations. Initial visits to the works and incoming pipes indicated a high degree of sediment deposition in the two inlet pipes. Analyses were carried out and consequently, changes to the hydraulic regime were made. Measurements of sediment level, sediment quality, wall slime and bulk water quality were monitored in the period following the remedial works to observe any improvements. Dramatic alterations in each of the determinands measured were recorded. Analyses were then undertaken to determine long term sediment behaviour and to assess the future usefulness of existing upstream sediment traps. It was concluded that with proper maintenance of the traps, the new hydraulic regime is sufficient to prevent further significant build up of sediment deposits and reduce impacts on the WTP. Further investigations made by North of Scotland Water Authority highlighted trade inputs to the system which may also have contributed to the now managed foul flush problem. PMID- 11902481 TI - Anoxic transformations of wastewater organic matter in sewers--process kinetics, model concept and wastewater treatment potential. AB - The sewer is an integral part of the urban wastewater system: the sewer, the wastewater treatment plant and the local receiving waters. The sewer is a reactor for microbial changes of the wastewater during transport, affecting the quality of the wastewater and thereby the successive treatment processes or receiving water impacts during combined sewer overflows. This paper presents the results of studies on anoxic processes, namely denitrification, in the bulk water phase of wastewater as it occurs in sewers. Experiments conducted on 12 different wastewater samples have shown that the denitrification process in the bulk wastewater can be simplified by the reduction of nitrate to nitrogen with significant accumulation of nitrite in the water phase. Utilization of nitrate was observed not to be limited by nitrate for concentrations above 5 gNO3-N/m3. The denitrification rates, under conditions of excess substrate and electron acceptor, were found to be in the range of 0.8-2.0 g NO3-N/(m3h). A discussion on the interaction of the sewer processes and the effects on a downstream located wastewater treatment plant (WWTP) is provided. PMID- 11902480 TI - Dissolved oxygen in gravity sewers--measurement and simulation. AB - Dissolved oxygen (DO) concentrations were during 2 months continuously measured in an intercepting sewer. Measurements were made upstream and downstream in a 3.6 km gravity sewer. DO showed significant diurnal variations mainly caused by changes in the organic matter composition of the wastewater. At low temperatures the gravity sewer was strictly aerobic. However, towards the end of the measuring campaign, DO concentrations decreased as temperature increased and the sewer became anaerobic part of the day. A conceptual model that takes into account bulk water and biofilm DO uptake as well as reaeration was used to simulate the DO measured. Using measurements from the upstream station as input, the model was calibrated to yield good validation results of the DO at the downstream station. PMID- 11902482 TI - Erosion mechanisms in combined sewers and the potential for pollutant release to receiving waters and water treatment plants. AB - The problems associated with solids in sewerage systems result in common difficulties such as blockages and flooding and the subsequent maintenance requirements have been well documented. Concerns regarding pollutant release have also been demonstrated, with the contribution from in-sewer solids to the quality of the flow during a storm event being especially significant. These events known as "foul flushes" in combined sewers typically occur in the initial period of storm flows, when the concentration of suspended sediments and other pollutants are significantly higher than at other times. Traditionally impacts from these events have been related to the suspended solids phase of the flow passing through a CSO structure. It is now apparent that much of the suspended load originates from solids eroded from the bed. The "near bed solids" which are re entrained into the flow, together with solids eroded from the bulk bed, account for large changes in the suspended sediment concentration under time varying flow conditions. The influence of these eroded solids and their potential impact on receiving waters and treatment plants will be reviewed using data obtained from field studies carried out in the main Dundee interceptor sewer in Scotland. This paper describes some of the methods employed to investigate the characteristics of the pollutants associated with solids erosion in combined sewers. PMID- 11902483 TI - Anaerobic transformations of wastewater organic matter and sulfide production- investigations in a pilot plant pressure sewer. AB - Anaerobic transformations of wastewater organic matter and sulfide production rate were studied using a pilot plant pressure sewer (inner diameter: 102 mm, length: 47 m). Furthermore, a process model description including carbon and sulfur cycle was presented. Wastewater characterization based on oxygen utilization rate (OUR) measurement and VFA analysis was employed. Under anaerobic conditions, a net production of readily biodegradable substrate was observed, which fact is important for biological removal of nitrogen and phosphorus at subsequent wastewater treatment plants. Model parameters were determined on the basis of experimental findings. The model simulation of transformations of organic matter in sewers can be used as input to the model simulation and evaluation of the processes in wastewater treatment plants. The model is also useful to evaluate the problems in both sewers themselves and treatment plants caused by hydrogen sulfide. PMID- 11902484 TI - Deterministic modelling of integrated urban drainage systems. AB - Today, the main concepts required for describing the dynamics of drainage in an entire urban area are known and models are available that can reasonably simulate the behaviour of the urban water system. Still, such integrated modelling is a complex exercise not only due to the sheer size of the model, but also due to the different modelling approaches that reflect the history of the sub-models used and of the purpose they were built for. The paper reviews the state of the art in deterministic modelling, outlines experiences and discusses problems and future developments. PMID- 11902485 TI - Analysis of scenarios for sewerage, wastewater treatment and prioritised load on environment from the Greater City of Copenhagen. AB - The sewer system for the Greater Copenhagen area covers an area of 4460 ha contributing to the runoff. The total area serves in total 8 municipalities, however it is dominated by the areas in the City of Copenhagen proper. The catchments merge into interceptors, which feed two large treatment plants. The effluent from the two treatment plants discharges during dry weather to Oresund, the sound between Denmark and Sweden. This large system has been analysed for selected scenarios with respect to handling runoff in an optimal way in order to minimise the loads on the most sensitive receiving waters and optimising treatment plant performance. PMID- 11902486 TI - Is peritoneal carcinomatosis an incurable disease or controllable locoregional condition?--Challenge of surgeons with intraperitoneal hyperthermic chemotherapy. PMID- 11902487 TI - Review of a personal experience in the management of carcinomatosis and sarcomatosis. AB - BACKGROUND: Peritoneal surface malignancy can result from seeding of gastrointestinal cancer or abdomino-pelvic sarcoma; it can also occur as a primary disease, such as peritoneal mesothelioma. In the past, this clinical situation was treated only with palliative intent. METHODS: An aggressive approach to peritoneal surface malignancy involves peritonectomy procedures, perioperative intraperitoneal chemotherapy and knowledgeable patient selection. The clinical assessments necessary for valid clinical judgements include the cancer histopathology (invasive vs expansive progression), the preoperative abdominal and pelvic CT, the peritoneal cancer index and the completeness of cytoreduction score. Proper patient selection is mandatory for optimizing the results of treatment. RESULTS: In a series of phase II studies, appendiceal tumors with peritoneal seeding became the paradigm for success with an 85% long term survival in selected patients. Carcinomatosis from colon cancer had an overall 5-year survival of 50% with selected patients. Also, sarcomatosis patients overall had a 40% 5-year survival in selected patients. Peritoneal mesothelioma showed a 36% 5-year survival. In all malignancies, early aggressive treatment of minimal peritoneal surface dissemination showed the greatest benefit. CONCLUSIONS: Oncologists must accept responsibility for knowledgeable management of peritoneal surface dissemination of cancer because a curative approach has been demonstrated in large phase II studies and all historical controls show 0% long-term survival. Adjuvant phase III studies with perioperative intraperitoneal chemotherapy in diseases where peritoneal surface spread occurs are indicated. PMID- 11902488 TI - Levels of soluble intercellular adhesion molecule-1 and total sialic acid in serum of patients with laryngeal cancer. AB - BACKGROUND: Adhesion molecules have been implicated in tumor progression. In this study, we aimed to investigate serum soluble intercellular adhesion molecule-1 (sICAM-1) and total sialic acid (TSA) levels in laryngeal carcinoma and correlate their levels with the cancer stage. METHOD: The sera from 35 patients with laryngeal cancer (10 at stage II, 12 at stage III and 13 at stage IV) were extracted before treatment. The concentrations of sICAM-1 and TSA were measured by enzyme-linked immunoassay and the thiobarbituric acid method, respectively and compared with those from a healthy control group (n = 34). RESULTS: Mean serum sICAM-1 and TSA levels were found to be higher in the total patient group (the lowest level belonging to stage II) than in the control group (p < 0.001, control versus total patient group). As the stage of the disease increased, higher levels of sICAM-1 and TSA were determined. The correlations between TSA and sICAM-1 became more significant as the stage of the disease increased (r= 0.67, p < 0.05 in stage II, r= 0.86, p < 0.001 in stage III and r = 0.90, p < 0.001 in stage IV). CONCLUSION: These data reveal that the significant correlations between sICAM-1 and TSA in laryngeal cancer, more prominent at advanced stage, might reflect the similar nature of these molecules, which function as adhesion molecules. PMID- 11902489 TI - Therapeutic results of alternating chemoradiotherapy for nasopharyngeal cancer using cisplatin and 5-fluorouracil: its usefulness and controversial points. AB - BACKGROUND: The present study was conducted to evaluate the therapeutic results of alternating chemoradiotherapy for locally advanced nasopharyngeal cancer (NPC). METHODS: The subjects consisted of six patients with stage III nasopharyngeal cancer and 26 patients with stage IV nasopharyngeal cancer. Using 6 MV photons, radiotherapy was performed at an exposure of 1.8-2.0 Gy five times per week. That is, a total absorbed dose of 36-40 Gy was irradiated between the base of the skull and supraclavicular fossa. After decreasing the irradiation field, an absorbed dose of 26-30 Gy was additionally given thereafter. One course of chemotherapy consisted of the administration of 5-fluorouracil (5-FU) at a dose of 700 mg/m2/24 h for 5 days (days 1-5) and cisplatin (CDDP) at a dose of 50 mg/m2/24 h for 2 days (days 6-7) and a total of 2-3 courses of chemotherapy were performed. During the alternating chemoradiotherapy, chemotherapy was performed initially and 3-5 days after completing the chemotherapy, radiotherapy was performed for 3-4 weeks. Thereafter, chemotherapy and radiotherapy were performed alternately. RESULTS: The scheduled courses of alternating chemoradiotherapy were completed in 30 (94%) of 32 patients. Although one patient developed shock induced by metal allergy to CDDP, no severe adverse effects were noted in any other patients. In these 32 patients, the overall 5-year survival rate was 75% (95% confidence interval: 60-90%) and the progression-free survival rate was 63% (95% CI: 46-89%). CONCLUSIONS: This method of alternating chemoradiotherapy yielded higher or at least similar survival rates and lower toxicities than concurrent chemoradiotherapy and is worth trying in a multi-institutional study. PMID- 11902490 TI - A phase I/II study of cisplatin and vinorelbine chemotherapy in patients with advanced non-small cell lung cancer. AB - BACKGROUND: A combination of cisplatin and vinorelbine chemotherapy is effective in cases of advanced non-small cell lung cancer, but the optimum administration schedule for both drugs has not yet been defined. The aim of this study was to determine the maximum dose of vinorelbine that can be tolerated while receiving a fixed dose of cisplatin every 3 weeks and to observe the response in Japanese patients with advanced non-small cell lung cancer who had not previously received chemotherapy. METHODS: Cisplatin was given at a dose of 80 mg/m2 on day 1. Vinorelbine was administered on days 1 and 8 at a starting dose of 25 mg/m2 that was then increased by 5 mg/m2 increments. This treatment was repeated every 3 weeks. RESULTS: Twenty-one patients received a total of 54 chemotherapy cycles consisting of three different vinorelbine dosages. Toxicity and efficacy were evaluated in all of the patients. The main dose-limiting toxicity was neutropenia. Grades 3-4 leukopenia and neutropenia were observed in 57% and 86% of all cycles, respectively. These conditions were reversible and did not result in death from toxicity. The most severe non-hematological toxicity symptom was a grade 3 infection and reaction at the site of injection. The maximum tolerated dose of vinorelbine was 35 mg/m2. The objective response was noted in one of six patients at dose level 1, in four of 12 patients at dose level 2 and in two of three patients at dose level 3. CONCLUSION: The recommended doses were 80 mg/m2 for cisplatin and 30 mg/m2 for vinorelbine. The combination of cisplatin and vinorelbine repeated every 3 weeks is well tolerated and has shown promising anti tumor activity against non-small cell lung cancer. PMID- 11902491 TI - Chemotherapy of thymic carcinoma: analysis of seven cases and review of the literature. AB - BACKGROUND: Thymic carcinoma has a dismal prognosis compared with thymoma, because most of such tumors have locoregional invasion when diagnosed. Thus the important step in the management of thymic carcinoma is the introduction of systemic chemotherapy. However, as thymic carcinoma is a rare neoplasm, treatment with chemotherapy has not been studied systematically. METHODS: We analyzed seven cases of primary thymic carcinoma, treated with various chemotherapy regimens in our hospital from 1990 to 1999, and carried out a literature review of case reports of thymic carcinoma successfully treated with chemotherapy. RESULTS: All four cases who received modified ADOC therapy obtained partial responses. Other chemotherapeutic regimens (CHOP-E, PVB) were not effective. CONCLUSION: Based on the results of this study and the literature review, we feel that a positive response is obtainable with chemotherapy for thymic carcinoma. Modified ADOC therapy showed consistent efficacy in thymic carcinoma in this study. PMID- 11902492 TI - Weekly high-dose 5-fluorouracil (5-FU), leucovorin (LV) and bimonthly cisplatin in patients with advanced gastric cancer. AB - BACKGROUND: A phase II clinical trial was performed to evaluate the activity and toxicity of bimonthly cisplatin and weekly 24-h infusion of high-dose 5 fluorouracil and leucovorin in patients with advanced gastric cancer. PATIENTS AND METHODS: From September 1997 to March 1998, 23 chemo-naive patients of advanced gastric cancer were enrolled in this study. The regimen consisted of weekly 24-h infusion of 5-FU (2,600 mg/m2) and LV 150 mg and bimonthly cisplatin (25-50 mg/m2) bolus for 12 weeks followed by a 2-week break. RESULTS: There were 10 male and 13 female patients with a median age of 52 years. A total of 428 chemotherapy treatments were given with a mean of 11. Seventeen patients were evaluable for response. There were 41% (7/17) partial response, 18% (3/17) stable disease and 41% (7/17) progressive disease. The grade III or IV toxicity included anorexia 35% (8/23), fatigue 26% (6/23), vomiting 17% (4/23) and mucositis 9% (2/23). One patient developed perforated duodenal stump after chemotherapy. One patient died of hyperammonemia-related coma. The median times to disease progression and overall survival were 3.5 and 7 months, respectively. CONCLUSIONS: This regimen showed modest activity against gastric cancer. However, there was no survival advantage and there was greater toxicity than with weekly high-dose 5-FU-LV alone. PMID- 11902493 TI - Phase II study of a weekly 8-hour 5-fluorouracil and leucovorin infusion for patients with advanced colorectal cancer: dose adjusted according to its toxicity. AB - BACKGROUND: 5-fluorouracil (5-FU) clearly behaves as two different drugs according to the schedules for its administration. A weekly, 8-h 5-FU continuous infusion (CI) regimen may produce a dual effect, because it elicits both a high plasma 5-FU level and also a durable exposure to 5-FU, which may have the advantage of inhibiting both DNA synthesis and RNA activities. The plasma 5-FU level, however, cannot be monitored in most hospitals, so we initiated a pragmatic clinical trial with this weekly 8-h 5-FU Cl regimen and adjusted the drug's dose according to the detected toxicity. METHODS: The initial dose of 5-FU was 1200 mg/m2 and this was escalated by 200 mg/m2 weekly, provided that no evidence of significant (grade 2 or greater) toxicity became apparent. Twenty-six patients entered the study from June 1998 to March 1999. RESULTS: The median dose of 5-FU delivered was 1600 mg/m2. The major symptoms precluding dose escalation were nausea and vomiting. Seven patients demonstrated a partial response (26.9%), 11 patients revealed stable disease (42.3%) and eight exhibited progressive disease (30.8%). CONCLUSION: This weekly 8-h CI 5-FU protocol with the adjustment of dose according to toxicity was not able to achieve the same 5-FU dose and response rate as in previous studies with pharmacokinetic monitoring of 5-FU levels. However, with the concurrent administration of intensive anti-emetic premedication, it is still possible to achieve adequate plasma 5-FU levels by adjusting the 5-FU dose according to elicited toxicity. PMID- 11902494 TI - Pancreas cancer incidence in the world. PMID- 11902495 TI - Confusion over mammography screening intensifies. PMID- 11902496 TI - Chemotherapy safe in pregnancy. PMID- 11902497 TI - Delhi launches cancer awareness campaign. PMID- 11902498 TI - Euthanasia: the Dutch experience and what it entails in practice. PMID- 11902500 TI - Human recombinant erythropoietin and quality of life: a wonder drug or something to wonder about? AB - Over the past decade an increasing number of studies have supported the use of recombinant human erythropoietin (epoetin) in cancer patients, suggesting that it improves haemoglobin concentrations for some. There is also evidence that this treatment may lead to improvement in quality of life for cancer patients. This systematic review examines the issue. We identified and critically reviewed 13 trials. Although some of the results indicate that epoetin has positive effects on quality of life, methodological limitations inherent in most of the studies hamper interpretation of data. Evidence from this review suggests that more robust designs are required to show any significant quality-of-life benefits for cancer patients undergoing epoetin treatment. PMID- 11902499 TI - Trastuzumab: hopes and realities. AB - Despite improvements in care of patients with breast cancer, up to half develop refractory or resistant disease. There is therefore a need for new, modified anticancer therapies with greater effectiveness, tolerability to patients, and tumour specificity. Trastuzumab (Herceptin) is the first clinically available oncogene-targeted therapeutic agent for treatment of solid tumours. Clinical trials in patients positive for HER2 (human epidermal-growth-factor receptor 2) show that trastuzumab is effective and well tolerated; as a single-agent second line or third-line treatment, the drug produced durable tumour responses. First line trastuzumab in combination with chemotherapy, particularly paclitaxel, significantly improved time to disease progression, duration of response, and time to treatment failure. Combination therapy resulted in a 25% improvement in overall survival compared with chemotherapy alone. Patients with HER2 gene amplification, high overexpression of HER2 (3+ on immunohisto-chemistry), or both features, obtained the greatest clinical benefit. Trastuzumab is the first monoclonal antibody with efficacy in breast cancer and the first gene-product targeted therapy to produce a significant survival advantage in this disease. Trastuzumab is likely to find its ultimate role in the adjuvant setting. Its development provides a model for the integration of other gene-targeted therapies into breast-cancer management to improve survival and quality of life. PMID- 11902501 TI - Myeloprotection with drug-resistance genes. AB - One of the many applications of gene transfer for cancer gene therapy is the transfer of drug-resistance genes into bone-marrow stem cells for myeloprotection. Protection of the hosts' bone marrow should allow for dose escalation that may be useful for eradicating minimal residual disease in a post transplant situation. A number of drug resistance genes, whose products include mutant forms of enzymes that confer resistance to chemotherapeutic drugs, are discussed. Advances in hematopoietic stem cell isolation and ex vivo manipulation has kept pace with improvements in retroviral vector technology to make hematopoietic stem cell transduction a distinct reality. Clinical trials, which have established that the approach is safe, are now being designed to address more therapeutically relevant issues. PMID- 11902503 TI - Nonsteroidal anti-inflammatory drugs, apoptosis, and colon-cancer chemoprevention. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) can inhibit colorectal tumorigenesis and are among the few agents known to be chemopreventive. Epidemiological studies and experiments with animals have shown that NSAIDs have powerful anticolorectal cancer properties, but the mechanism of these effects remains unclear. NSAIDs can inhibit neoplastic growth by inducing apoptosis in cancer cells; the way they do this is currently an area of intense investigation. The most well-characterised pharmacological feature of NSAIDs is their inhibition of the enzyme cyclo-oxygenase (COX), which catalyses the synthesis of prostaglandins. Several studies have shown that COX inhibition prevents cell proliferation and promotes apoptosis. The chemopreventive effects of NSAIDs are thought to occur via this pathway. Other observations indicate that NSAIDs also promote apoptosis through mechanisms that are independent of COX inhibition. This idea is supported by the finding that compounds that are structurally similar to NSAIDs, but do not inhibit COX, also have chemopreventive and proapoptotic properties. COX-dependent and COX-independent mechanisms of apoptosis induction are not mutually exclusive, and it is likely that both have a role in the biological activity of NSAIDs. Knowledge of how NSAIDs prevent neoplastic growth will greatly aid the design of better chemopreventive drugs and novel treatments for colorectal cancer. PMID- 11902502 TI - Diagnostic accuracy of dermoscopy. AB - The accuracy of the dinical diagnosis of cutaneous melanoma with the unaided eye is only about 60%. Dermoscopy, a non-invasive, in vivo technique for the microscopic examination of pigmented skin lesions, has the potential to improve the diagnostic accuracy. Our objectives were to review previous publications, to compare the accuracy of melanoma diagnosis with and without dermoscopy, and to assess the influence of study characteristics on the diagnostic accuracy. We searched for publications between 1987 and 2000 and identified 27 studies eligible for meta-analysis. The diagnostic accuracy for melanoma was significantly higher with dermoscopy than without this technique (log odds ratio 4.0 [95% CI 3.0 to 5.1] versus 2.7 [1.9 to 3.4]; an improvement of 49%, p = 0.001). The diagnostic accuracy of dermoscopy significantly depended on the degree of experience of the examiners. Dermoscopy by untrained or less experienced examiners was no better than clinical inspection without dermoscopy. The diagnostic performance of dermoscopy improved when the diagnosis was made by a group of examiners in consensus and diminished as the prevalence of melanoma increased. A comparison of various diagnostic algorithms for dermoscopy showed no significant differences in their diagnostic performance. A thorough appraisal of the study characteristics showed that most of the studies were potentially influenced by verification bias. In conclusion, dermoscopy improves the diagnostic accuracy for melanoma in comparison with inspection by the unaided eye, but only for experienced examiners. PMID- 11902504 TI - Mediastinal lymphoadenopathy in a patient with breast cancer. PMID- 11902505 TI - Quality-of-life assessment in palliative care. AB - Success in oncology has traditionally been measured in terms of cure, survival, and tumour response. However, more recently, health-related quality of life has emerged as an important outcome, particularly in the palliation setting. We review published randomised studies from two areas in palliation: those assessing the effectiveness of palliative care programmes and those looking at the effects of palliative chemotherapy compared with best supportive care. In the latter studies, there was an improvement in research methods between the late 1980s and 2000, owing to the use of standardised instruments, specification of endpoints, and improvements in data presentation and interpretation. A range of health related quality-of-life instruments were used in the studies, which makes comparisons difficult. This was particularly true of the palliative-care programmes. Attrition due to the death of patients in the study groups was also a problem and needs to be taken into account in study planning and design, as well as in data collection. A common standard for scoring health-related quality of life measurements both within and between instruments would improve the interpretation of findings and their clinical application, thereby giving them greater effect on clinical decision-making. PMID- 11902506 TI - Peter Cardy--MacMillan Cancer Relief. PMID- 11902507 TI - Theory and practice. PMID- 11902508 TI - Poor cancer-screening uptake among ethnic minorities. PMID- 11902509 TI - Use of new cancer drugs in India. PMID- 11902510 TI - Malignant disease and von Willebrand factor. PMID- 11902511 TI - Procaspase activation in lung carcinomas. PMID- 11902512 TI - The changing role of UK primary cancer care. PMID- 11902513 TI - Variations in treatment and survival in breast cancer. AB - To achieve optimum quality of care for women with breast cancer in the UK, uniformity of care in accordance with consensus guidelines is needed. This review highlights variations in provision of care for women with breast cancer, with particular emphasis on care received in the UK, examines differences in survival, and discusses the factors that may underlie these differences. Strong variation in treatment was identified, which appeared to affect survival significantly. These findings reinforce the need for women with breast cancer to be treated by dedicated specialists working within a multidisciplinary team to provide a high standard of care. PMID- 11902514 TI - Angiogenesis as a biomarker and target in cancer chemoprevention. AB - Angiogenesis, the formation of new blood vessels from existing vasculature, is essential to the late stages of carcinogenesis, allowing tumours to grow beyond 1 2 mm in diameter, invade surrounding tissue, and metastasise. However, more than two decades ago, angiogenesis that preceded neoplastic transformation was seen. Indeed, it can be detected in inflammatory and infectious diseases that increase the risk of developing cancer. Recent advances in fluorescence endoscopy and histological assessment suggest that, for certain cancers, the degree of new blood-vessel formation may differ between the early and late stages of carcinogenic progression. The association between angiogenesis and cancer occurrence, and ease of detection of this process in accessible tissues early in carcinogenesis, mean that angiogenesis fulfils the criteria for a biomarker of the effectiveness of chemopreventive intervention. There is also some evidence that biochemical assays of angiogenic growth factors may offer similar potential as surrogate biomarkers. Many natural and synthetic chemopreventive agents in development or in clinical use inhibit new vessel formation in vivo. Validation of angiogenesis as a biomarker for the effectiveness of chemoprevention should further the advancement of some chemopreventive agents. PMID- 11902515 TI - Metronomic scheduling: the future of chemotherapy? AB - Tumour endothelium is a new target for anticancer treatments. Proliferating endothelial cells from the tumour, even if qualitatively different from those of blood vessels in the normal tissue of origin, remain putatively normal and genetically stable cells. The results of recent experimental studies have suggested that frequent administration of certain cytotoxic agents at low doses (a tenth to a third of the maximum tolerated dose), known as 'metronomic' chemotherapy, increases the antiangiogenic activity of the drugs. The effects of these metronomic schedules of cytotoxic agents may be further enhanced by concurrent administration of novel, selective, treatments that inhibit, at a molecular level, the processes of tumour formation and growth eg angiogenesis, growth factor pathways, and other signal transduction cascades. The need to treat patients for long periods also supports the use of metronomic scheduling for chemotherapy, to minimise toxicity and to target both proliferating tumour cells and endothelial cells. This review describes the experimental studies involving metronomic schedules of chemotherapy, alone and in combination with angiogenesis inhibitors, and suggests a new therapeutic anticancer paradigm for controlling cancer by long-term therapy, based on the development of combinations of metronomic cytotoxic agents with individually tailored compounds designed to target specific molecules. PMID- 11902516 TI - No randomised trial of prostate-cancer screening in Norway. AB - The Norwegian Urological Cancer Group (NUCG) decided against participating in the European Randomised Study on Screening for Prostate Cancer, to which they had planned to contribute data from a population of 75 000 men aged between 50 and 65. The plan was abandonned for three main reasons: Norwegian doubts about the statistical power of the trial, the possible ethical implications of screening a large number of men with no symptoms, and the belief that the trial would not produce clinically important results. In this review, the NUCG's reasons for not participating in the trial are discussed in full. PMID- 11902517 TI - Dealing with stress, burnout, and grief in the practice of oncology. AB - There is a substantial amount of published work describing how to help patients and families deal with stress, grief, and loss, but there is much less advice available for clinicians dealing with similar issues. Working as an oncologist is inherently difficult, and racked by emotional and psychological traumas. However, it can also be tremendously rewarding. In this review, I discuss the extent of stress and burnout in the practice of oncology, its causes and manifestations, and strategies for overcoming it. PMID- 11902518 TI - Abortion and breast cancer: a hard decision made harder. AB - Over recent years, concerns have been raised about a possible causal relation between induced abortion and subsequent breast cancer. The abrupt hormonal changes associated with termination of pregnancy may induce changes in breast epithelial cells at a stage when they are not fully differentiated and therefore more vulnerable to later development of breast cancer. This review examines the published evidence supporting and refuting this hypothesis and concludes that there are, to date, insufficient data to justify warning women of future breast cancer risk when counselling them about abortion. PMID- 11902519 TI - Futile care in oncology: when to stop trying. PMID- 11902520 TI - Steven Pantilat: a palliative care specialist. (Interview by Barbara Boughton). PMID- 11902521 TI - Dealing with uncertainty. PMID- 11902523 TI - Mariano Barbacid: director of Spain's first national cancer centre. Interviewed by Adrian Burton. PMID- 11902522 TI - Paediatric oncology in Argentina: medical and ethical issues. AB - The process of globalisation is affecting health and health care in Argentina, as it is in many other countries. The full extent of this effect is still unclear, but winners and losers in the world economy are emerging--not only different countries, but also sectors or populations within those countries. There are serious inequalities in health-care provision in Argentina, so that not all children with cancer receive the best possible therapy. What happens to those children who don't? How do staff feel when they have to turn away new patients? Only by asking these questions and examining and understanding the answers can we begin the process of improving the status of paediatric oncology in Argentina. PMID- 11902524 TI - A paradox of progress. PMID- 11902526 TI - Belgian Parliament calls for organised cervical cancer screening and HPV research throughout Europe. PMID- 11902525 TI - Critical times for Argentinian cancer patients. PMID- 11902527 TI - Prediction of the response of colorectal cancer to systemic therapy. AB - Adjuvant chemotherapy with fluorouracil and folinic acid improves overall survival for resected carcinoma of the colon of Dukes' stage C by 10-12%. In metastatic disease, response rates with fluorouracil-based regimens are about 25%. Combination with newer agents such as irinotecan and oxaliplatin can improve response rates to more than 50% in selected patients. New treatments with novel molecular targets will soon be entering clinical use. Despite these improvements, many patients undergo chemotherapy for resistant cancer, thus incurring side effects without benefit. Expression of particular genes can be tested at the protein or RNA level and can be correlated with response or resistance to particular systemic therapies. Thus, predictive-factor testing of tumour biopsy samples may allow us to select chemotherapy or immunotherapy treatments with a high likelihood of benefit for the individual patient. PMID- 11902528 TI - The biological treatment of renal-cell carcinoma and melanoma. AB - Biological therapies are claiming a place in the routine management of some solid tumours. In this review we focus on the biological treatment of melanoma and renal-cell carcinoma, identifying the background to current practice and areas of promise that may be in routine clinical use in the near future. Melanomas and renal-cell carcinomas are particularly resistant to chemotherapy and radiotherapy and are characterised by the host immune response to the tumours. For this reason there has been particular interest in the biological therapy of these diseases. Biological therapies differ from chemotherapeutic approaches in their mechanism of action, time to response, and side-effect profiles. Although biological treatment has a long history, it is only with recent advances in immunology and molecular biology that progress has been made. In the next few years investigators expect to build on their research experience with biotherapeutic agents to provide tangible benefits for patients. PMID- 11902529 TI - Gastric MALT lymphoma: from aetiology to treatment. AB - The development of gastric mucosa-associated lymphoid tissue (MALT) lymphoma is dependent on Helicobacter pylori infection. Bacterial colonisation of the gastric mucosa triggers lymphoid infiltration and the formation of acquired MALT. The bacterial infection induces and sustains an actively proliferating B-cell population through direct (autoantigen) and indirect (intratumoral T cells specific for H. pylori) immunological stimulation. Moreover, the bacterial infection provokes a neutrophilic response, which causes the release of oxygen free radicals. These reactive species may promote the acquisition of genetic abnormalities and malignant transformation of reactive B cells. A transformed clone carrying the translocation t(1;18)(q21;q21) forms a MALT lymphoma, the growth of which is independent of H. pylori and will not respond to bacterial eradication. Malignant clones without t(11;18)(q21;q21), but with other genetic abnormalities, such as trisomy 3 or microsatellite instability, depend critically on immune stimulation mediated by H. pylori for their clonal expansion. In the early stages, the tumour can be successfully treated by eradication of the bacterium, whereas at later stages the tumour may escape its growth dependency through acquisition of additional genetic abnormalities such as t(1;14)(p22;q32) and t(1;2)(p22,p12) involving the BCL-10 gene. Finally, further genetic abnormalities, such as inactivation of the tumour suppressor genes, p53 and p16, can lead to high-grade transformation. Detection of these abnormalities may help with the clinical management of patients with gastric MALT lymphoma. PMID- 11902530 TI - The changing face of UK primary cancer care. PMID- 11902531 TI - Japan plans new cancer screening system. PMID- 11902533 TI - Experts dissatisfied with cancer care in Germany. PMID- 11902532 TI - FISHy decisions in breast cancer treatment choice. PMID- 11902534 TI - National prostate tumour bank launched in Australia. PMID- 11902535 TI - An ID card for tumour cell lines: HLA typing can help. PMID- 11902536 TI - Positron emission tomography in the diagnosis and staging of lung cancer: a systematic, quantitative review. AB - Lung cancer is the cause of 32% of all male cancer deaths and 25% of all female cancer deaths. Because the prognosis depends on early diagnosis and staging, continuous evaluation of the diagnostic tools available is important. The aim of this study was to assess the diagnostic value of dedicated positron emission tomography (PET) and gamma-camera PET in the diagnostic investigation of non small-cell lung cancer (NSCLC). A systematic literature search was carried out in the MEDLINE and EMBASE databases and the Cochrane Controlled Trials Register. We identified 55 original works on the diagnostic performance of PET with fluorodeoxyglucose in the investigation of NSCLC. For diagnosis of NSCLC, the mean sensitivities and specificities were, respectively, 0.96 (SE 0.01) and 0.78 (0.03) for dedicated PET, and 0.92 (0.04) and 0.86 (0.04) for gamma-camera PET. In the mediastinal staging of NSCLC, the results were 0.83 (0.02) and 0.96 (0.01) for dedicated PET and 0.81 (0.04) and 0.95 (0.02) for ganuna-camera PET. We conclude that dedicated PET could be a valuable tool in the diagnosis and staging of NSCLC. However, studies of populations with a lower prevalence of NSCLC are recommended. PMID- 11902537 TI - Vascular endothelial growth factor: its prognostic, predictive, and therapeutic implications. AB - Since the discovery that cancer development requires the growth of new blood vessels, many investigations have revealed the key molecules in the regulation of new vessel formation. One of the most important of these molecules is vascular endothelial growth factor (VEGF)--an endothelial-cell-specific mitogen and survival factor. VEGF also causes increased vascular permeability and recruits progenitor endothelial cells from the bone marrow. Clinical observations have confirmed that VEGF status is closely associated with the extent of neovascularisation and prognosis in many solid tumours. VEGF status is predictive of resistance to various treatments, including radiotherapy, chemotherapy, and endocrine therapy. Preliminary results also indicate that anti-VEGF treatment suppresses cancer progression without serious toxic effects. Various approaches for the control of cancers involving inhibition of the activity of VEGF are currently being investigated. This review considers the clinical implications of VEGF, particularly its prognostic, predictive, and therapeutic value. PMID- 11902538 TI - Gene expression profiling of cancer by use of DNA arrays: how far from the clinic? AB - DNA arrays allow the simultaneous analysis of expression levels for thousands of genes in normal and pathological tissues and hold great promise in molecular medicine, notably in cancer research. The great biological and clinical diversity present in human tumours is poorly characterised by the current classification systems. DNA arrays can provide a better understanding of oncogenesis, leading to improvements in cancer management. First, the identification of new target genes and pathways will allow the development of specific molecular-based anticancer drugs. Secondly, expression profiles will permit tumour classification in more homogeneous diagnostic and prognostic groups, as well as the identification of new clinically and biologically relevant tumour subclasses. Here, we review the technology and present some cancer studies with promising results. Finally, we discuss some of the issues that must be resolved in the near future, so that DNA arrays can fulfil the aims mentioned above. PMID- 11902539 TI - Esthesioneuroblastoma: a meta-analysis and review. AB - Our objective was to review recent developments in diagnosis, staging, and treatment of esthesioneuroblastoma (ENB). A meta-analysis of publications between 1990 and 2000 was carried out, and studies were classified according to their main subject: origin/aetiology of ENB, histopathological diagnosis, and treatment. Data so far point to the basal progenitor cells of the olfactory epithelium as the origin of ENB. Histopathological diagnosis remains difficult and is based on results of antigen expression detected through a panel of antibodies by immunohistochemistry. RT-PCR of HASH expression could be a specific marker of ENB. Overall and disease-free survival at 5 years averaged 45% (SD 22) and 41% (SD 21) in the studies included in the meta-analysis. Survival in Hyams' grades I-II was 56% (SD 20) compared with 25% (SD 20) in grades III-IV (odds ratio 6.2). In patients with metastases in cervical lymph nodes (on average 5% of the total) survival was 29%, compared with 64% for patients with N0 disease (odds ratio 5.1). Survival according to treatment modalities was 65% for surgery plus radiotherapy, 51% for radiotherapy and chemotherapy, 48% for surgery, 47% for surgery plus radiotherapy and chemotherapy, and 37% for radiotherapy alone. The histopathological grading according to Hyams and the presence of cervical lymph node metastases emerged as prognostic factors. A combination of surgery and radiotherapy seems to be the optimum approach to treatment. The exact role of chemotherapy in treatment protocols is still unclear. The role of elective neck dissection is unclear. PMID- 11902540 TI - Patients' preferences for adjuvant chemotherapy in early breast cancer: a review of what makes it worthwhile. AB - Adjuvant chemotherapy for early breast cancer improves survival but is unpleasant and inconvenient. Women and clinicians need information about the magnitude of survival benefits considered sufficient to make the side-effects and inconvenience worthwhile. We sought studies that quantified the minimum required survival benefit by asking women who had experienced adjuvant chemotherapy. Four studies involving 512 women were appraised and summarised. All studies referred to chemotherapy given between 1980 and 1996, but the methods varied widely, especially the way in which women were recruited and questioned. However, the results were remarkably consistent. Most women said that small improvements in survival were sufficient to make adjuvant chemotherapy worthwhile. Women with dependants, social support, and milder side-effects judged smaller benefits worthwhile. Age, education, employment, and income status were not predictive of the women's responses. Optimum decision-making about the relative benefit of adjuvant chemotherapy requires up-to-date information from women receiving current chemotherapy and supportive regimens. PMID- 11902541 TI - Trends in biomarker research for cancer detection. AB - A key challenge in cancer control and prevention is detection of the disease as early as possible, enabling effective interventions and therapies to contribute to reduction in mortality and morbidity. Biomarkers are important as molecular signposts of the physiological state of a cell at a specific time. Active genes, their respective protein products, and other organic chemicals made by the cell create these signposts. As a normal cell progresses through the complex process of transformation to a cancerous state, biomarkers could prove vital for the identification of early cancer and people at risk of developing cancer. We discuss current research into the genetic and molecular signatures of cells, including microsatellite instability, hypermethylation and single-nucleotide polymorphisms. The use of genomic and proteomic high-throughput technology platforms to facilitate detection of early cancer by means of biomarkers, and issues on the analysis, validation, and predictive value of biomarkers based on these technologies are also discussed. We report on recent advances in identifying sources of biomarkers that can be accessed by noninvasive techniques, such as buccal-cell isolates, as well as traditional sources such as serum or plasma. We also focus on the work of the Early Detection Research Network at the National Cancer Institute, harnessing expertise from leading national and international institutions, to identify and validate biomarkers for the detection of precancerous and cancerous cells in assessing risk of cancer. The network also has a role in linking discovery to process development, resulting in early detection tests and clinical assessment. PMID- 11902542 TI - Left atrial sarcoma presenting as cerebral infarction. PMID- 11902543 TI - What price specialisation? PMID- 11902544 TI - Learning from the literature. PMID- 11902545 TI - Clinical research comes under scrutiny. PMID- 11902546 TI - Vitamine D derivatives convert colon cancer cells. PMID- 11902547 TI - Mosaic tumour blood vessels and high-dose chemotherapy for breast cancer. PMID- 11902548 TI - The unresolved crisis in clinical research can be resolved. PMID- 11902549 TI - Childhood acute lymphoblastic leukaemia--current status and future perspectives. AB - The current cure rate of 80% in childhood acute lymphoblastic leukaemia attests to the effectiveness of risk-directed therapy developed through well-designed clinical trials. In the past decade there have been remarkable advances in the definition of the molecular abnormalities involved in leukaemogenesis and drug resistance. These advances have led to the development of promising new therapeutic strategies, including agents targeted to the molecular lesions that cause leukaemia. The importance of host pharmacogenetics has also been recognised. Thus, genetic polymorphisms of certain enzymes have been linked with host susceptibility to the development of de novo leukaemia or therapy-related second cancers. Furthermore, recognition of inherited differences in the metabolism of antileukaemic agents has provided rational selection criteria for optimal drug dosages and scheduling. Treatment response assessed by measurements of submicroscopic leukaemia (minimal residual disease) has emerged as a powerful and independent prognostic indicator for gauging the intensity of therapy. Ultimately, treatment based on biological features of leukaemic cells, host genetics, and the amount of residual disease should improve cure rates further. PMID- 11902550 TI - Intramedullary spinal cord metastasis. PMID- 11902551 TI - The role of the coagulation system in tumour angiogenesis. AB - The coagulation system, which is activated in most cancer patients, has an important role in tumour biology. It may make a substantial contribution to tumour angiogenesis, which represents an imbalance in the normal mechanisms that allow organised healing after injury. The recently recognised, but steadily growing, knowledge of the relationship between the coagulation and angiogenesis pathways has research and clinical implications. Manipulation of these systems may minimise both the neoangiogenesis essential for tumour growth and associated thromboembolic complications. However, since surgery is the primary treatment for most cancers, the angiogenesis of wound healing and haemostatic competence must be maintained. In this article, we summarise the complex interactions between the coagulation system and the angiogenic process that occur in cancer growth. We focus upon the contributions of the vascular endothelium, platelets, and coagulation factors to the angiogenic process and explore the coagulation system as a therapeutic target. PMID- 11902552 TI - Lymph-node metastases in patients with melanoma: what is the optimum management? AB - Lymph-node metastasis is an indicator of poor prognosis for patients with melanoma. The management of regional nodes is controversial, with continuing debate about whether surgery or radiotherapy of positive lymph nodes improves long-term survival or whether nodal involvement is merely a marker of aggressive disease. However, there is general agreement that systemic chemotherapy is rarely an effective form of management. This review therefore considers surgical and radiotherapeutic aspects of lymph-node management in patients with melanoma. We discuss regional control and survival after lymph-node surgery in retrospective series, randomised trials of elective lymph-node dissection, the role of 'sentinel' lymph-node biopsy, radiobiology and radiotherapy fractionation issues in melanoma treatment, retrospective studies of adjuvant nodal radiotherapy, and finally, randomised trials of adjuvant radiotherapy after lymph-node dissection. PMID- 11902553 TI - Antiviral approaches for cancers related to Epstein-Barr virus and human papillomavirus. AB - Epstein-Barr virus and human papillomaviruses (HPV) are DNA viruses underlying the carcinogenesis of 15-20% of human cancers worldwide. Viral oncoproteins are involved in malignant transformation and maintenance of the malignant phenotype, mainly through interaction between oncoproteins and products of tumour-suppressor genes. The use of vaccines to prevent the occurrence of HPV-related cancers is being investigated. Several approaches have been used to inhibit expression of viral oncoproteins. The first strategy uses antisense oligodeoxynucleotides against viral oncoproteins; downregulation of the oncoproteins can influence tumour cell growth and restore sensitivity to cytotoxic agents. Another approach uses antiviral drugs such as acyclic nucleoside phosphonates; inhibition of virus replication can lead to downregulation of viral oncoproteinsand ultimately reactivate tumour-suppressor-gene pathways. In addition, the combination of acyclic nucleoside phosphonates with conventional cytotoxic agents is more effective than either agent alone. These data provide the basis for a novel anticancer strategy to improve the therapeutic ratio in virus-related cancers, which needs to be further investigated for clinical applications. PMID- 11902554 TI - One hundred years of radiotherapy in Turkey. AB - The study and practice of radiology in Turkey began in 1897, only 2 years after the discovery of X-rays. A simple X-ray machine was constructed in Istanbul, consisting of a Crookes tube, a Ruhmkorff coil, and a home-made battery. This machine was first used on wounded soldiers, for diagnostic purposes. The first report of X-rays being used therapeutically in Turkey was published in a national journal in 1904. By 1933, the most up-to-date radiotherapy equipment of the time had been installed in every major city in the country. Innovative radiotherapy techniques, such as rotational treatment, were also being tried in 1930s. Today, there are 45 radiotherapy centres in Turkey, and 400 radiation oncologists and 80 medical physicists practise there. PMID- 11902555 TI - Treatment of metastatic melanoma in the brain with temozolomide and thalidomide. PMID- 11902557 TI - Alfred Knudson and his two-hit hypothesis. (Interview by Ezzie Hutchinson). PMID- 11902556 TI - Cancer care: what are the priorities? PMID- 11902558 TI - Hidden dangers. PMID- 11902559 TI - India's new smoking laws--progress or politics? PMID- 11902560 TI - Hatching headless chickens from rational eggs. PMID- 11902561 TI - Embryonic stem cells. PMID- 11902562 TI - Improving cancer care at the end of life. PMID- 11902563 TI - Epidemiology of breast cancer. AB - Breast cancer is the commonest cause of cancer death in women worldwide. Rates vary about five-fold around the world, but they are increasing in regions that until recently had low rates of the disease. Many of the established risk factors are linked to oestrogens. Risk is increased by early menarche, late menopause, and obesity in postmenopausal women, and prospective studies have shown that high concentrations of endogenous oestradiol are associated with an increase in risk. Childbearing reduces risk, with greater protection for early first birth and a larger number of births; breastfeeding probably has a protective effect. Both oral contraceptives and hormonal therapy for menopause cause a small increase in breast-cancer risk, which appears to diminish once use stops. Alcohol increases risk, whereas physical activity is probably protective. Mutations in certain genes greatly increase breast-cancer risk, but these account for a minority of cases. PMID- 11902564 TI - Mantle-cell lymphoma. AB - During the past decade, mantle-cell lymphoma has been established as a new disease entity. The normal counterparts of the cells forming this malignant lymphoma are found in the mantle zone of the lymph node, a thin layer surrounding the germinal follicles. These cells have small to medium-sized nuclei, are commonly indented or cleaved, and stain positively with CD5, CD20, cyclin D1, and FMC7 antibodies. Because of its morphological appearance and a resemblance to other low-grade lymphomas, many of which grow slowly, this lymphoma was initially thought to be an indolent tumour, but its natural course was not thoroughly investigated until the 1990s, when the BCL1 oncogene was identified as a marker for this disease. Mantle-cell lymphoma is a discrete entity, unrelated to small lymphocytic or small-cleaved-cell lymphomas. PMID- 11902565 TI - Role of nitric oxide in carcinogenesis and tumour progression. AB - Nitric oxide (NO) is a short-lived molecule required for many physiological functions, produced from L-arginine by NO synthases (NOS). It is a free radical, producing many reactive intermediates that account for its bioactivity. Sustained induction of the inducible form of NOS (iNOS) in chronic inflammation may be mutagenic, through NO-mediated DNA damage or hindrance to DNA repair, and thus potentially carcinogenic. Expression of iNOS is positively associated with P53 mutation in tumours of the colon, lung, and oropharynx. Progression of a large majority of human and experimental tumours seems to be stimulated by NO resulting from activation of iNOS or constitutive NOS, whereas inhibition is documented in others. This discrepancy is largely explained by differential sensitivity of tumour cells to NO-mediated cytostasis or apoptosis and clonal evolution of NO resistant and NO-dependent cells. P53 mutation or loss is one of many events linked with NO resistance and dependence. NO can stimulate tumour growth and metastasis by promoting migratory, invasive, and angiogenic abilities of tumour cells, which may also be triggered by activation of cyclo-oxygenase (COX)-2. Thus, selective inhibitors of NOS, COX, or both may have a therapeutic role in certain cancers. PMID- 11902566 TI - Clinical role of positron emission tomography in oncology. AB - Positron emission tomography (PET) is now in routine use in oncology, through the success of metabolic imaging, mainly with fluorodeoxyglucose (FDG). Clear benefit is obtained with FDG PET in the assessment of patients with recurrent or residual disease, especially colorectal cancer and lymphoma. Preoperative staging of non small-cell lung cancer with FDG PET is of proven benefit. Staging and restaging of patients with melanoma of stage II or greater is useful, and FDG PET has also been successfully used to investigate single pulmonary nodules. Tumour grading has been assessed, especially in the brain, but an important and emerging indication is the evaluation of tumour response with PET. Rapid decline of FDG uptake has been observed in responsive cancers. Further advances are being made with other fluorine-18-labelled and generator-based PET tracers, the only ones that can be used in units without dedicated cyclotrons. PMID- 11902567 TI - Treatment of cancer pain with transdermal fentanyl. AB - Pain is a feature of many cancers, particularly in the advanced stages at which the palliative care approach to symptom control achieves the best outcomes. The holistic approach generally dictates that any treatment of the cancer per se has symptom control as the primary objective at this advanced stage. Pain, which invariably increases with disease progression, is treated with opioids and adjuvant analgesic drugs together with physical therapies. Orally administered opioid drugs are used preferentially because of cost and convenience, but other routes of administration (subcutaneous, rectal, spinal) are also possible. More recently, transdermal fentanyl has been evaluated in the treatment of moderate to severe cancer pain. The rate of fentanyl absorption is constant (after a lag period), and the dose is altered by increasing or decreasing the area of skin covered by the patch (size and/or number of patches). The dosing interval for these systems is generally 3 days. The extent of pain relief provided by transdermal fentanyl and sustained release morphine formulations is similar, with quality-of-life instruments showing no consistent preference for either formulation. Open studies have suggested a lower risk of constipation. Transdermal fentanyl is effective in the treatment of severe cancer pain, particularly when the oral route is unavailable. PMID- 11902568 TI - Asymptomatic patient with an increasing concentration of CEA. PMID- 11902569 TI - 'Best supportive care' has had its day. AB - The term 'best supportive care' has been used since 1988 to describe the control arm in trials evaluating chemotherapy in advanced cancers. There are problems with this term. First, it implies that we more strenuously optimise these components of care in trials than in routine oncology practise, when there is no evidence that we do. Secondly, the name implies that it is effective, when usually it is not. Finally, 'best' suggests that we have reached a clear landmark of progress in non-chemotherapeutic palliation, when clearly we have not. 'Best supportive care' is an unhelpful and misleading term which should be avoided. In future trials, it should be replaced by 'standard palliative care ' with the type and frequency of key palliative interventions documented. PMID- 11902570 TI - Richard Wooster on cancer and the Human Genome Project (interview by Ezzie Hutchinson). PMID- 11902571 TI - Assisted suicide and cancer. PMID- 11902572 TI - Managing the managers. PMID- 11902573 TI - The promise of retinoids to fight against cancer. AB - Retinoids have a reputation for being both detrimental and beneficial: they are teratogens, but they also have tumour-suppressive capacity. Cell biology and genetics have significantly improved our understanding of the mechanisms that underlie the anti-proliferative action of retinoids. Recent elucidation of the pathways that are activated by retinoids will help us to exploit the beneficial aspects of this powerful class of compounds for cancer therapy and prevention. PMID- 11902575 TI - Cell senescence and cancer. AB - Historically, the senescent state has been associated with, and was named after, the cell-cycle arrest that occurs after cells have undergone an intrinsically defined number of divisions in vitro. More recently, however, it has been shown that extrinsic factors, including those encountered in normal tissue-culture environments, can prematurely induce an indistinguishable senescent phenotype. In this review, we discuss the pathways of cell senescence, the mechanisms involved and the role that these pathways have in regulating the initiation and progression of cancer. PMID- 11902574 TI - Histone deacetylases and cancer: causes and therapies. AB - Together, histone acetyltransferases and histone deacetylases (HDACs) determine the acetylation status of histones. This acetylation affects the regulation of gene expression, and inhibitors of HDACs have been found to cause growth arrest, differentiation and/or apoptosis of many tumours cells by altering the transcription of a small number of genes. HDAC inhibitors are proving to be an exciting therapeutic approach to cancer, but how do they exert this effect? PMID- 11902576 TI - FRA3B and other common fragile sites: the weakest links. AB - In 1979, the first chromosome alteration associated with familial cancer was reported. Five years later, a fragile site was observed in the same chromosome region. The product of the fragile histidine triad (FHIT) gene, which encompasses this fragile site, is partially or entirely lost in most human cancers, indicating that it has a tumour-suppressor function. Inactivation of only one FHIT allele compromises this suppressor function, indicating that a 'one-hit' mechanism of tumorigenesis is operative. Are genes disrupted at other fragile sites? And, are these genes also tumour suppressors? PMID- 11902577 TI - To cycle or not to cycle: a critical decision in cancer. AB - Tumour cells undergo uncontrolled proliferation, yet tumours most often originate from adult tissues, in which most cells are quiescent. So, the proliferative advantage of tumour cells arises from their ability to bypass quiescence. This can be due to increased mitogenic signalling and/or alterations that lower the threshold required for cell-cycle commitment. Understanding the molecular mechanisms that underlie this commitment should provide important insights into how normal cells become tumorigenic and how new anticancer strategies can be devised. PMID- 11902578 TI - Assessing TP53 status in human tumours to evaluate clinical outcome. AB - TP53 is probably the most extensively studied tumour-suppressor gene, and patients with TP53 mutations are known to have a poor outcome. However, inconsistencies in the analysis of TP53 status, and failure to realize that different mutations behave in different ways, prevent us from effectively applying our vast knowledge of this protein in clinical practice. What simple steps can be taken to ensure that patients benefit from our understanding of TP53? PMID- 11902579 TI - Comprehensive cancer centres and the war on cancer. AB - Comprehensive Cancer Centres are now recognized as an important weapon in the war on cancer, but they had to fight a very different battle to become accepted by the academic community. Why were these centres developed? How do they contribute to cancer research? Have they achieved the aims for which they were set up? And how should they be improved? It is important to answer these questions because we believe that cancer centres, though in need of improvement, are vital parts of our anticancer strategy. PMID- 11902580 TI - Chromosome translocations: dangerous liaisons revisited. AB - Although it has been clear for more than a century that the chromosomes in human tumour cells are often wildly abnormal, there has been controversy as to whether these changes are primary events or are merely secondary epiphenomena that reflect the genomic instability of these cells. The prevailing view for most of this period was that chromosome changes were secondary events. What happened to change this view? PMID- 11902581 TI - Scaling down imaging: molecular mapping of cancer in mice. AB - The development of miniaturized imaging equipment and reporter probes has improved our ability to study animal models of disease, such as transgenic and knockout mice. These technologies can now be used to continuously monitor in vivo tumour development, the effects of therapeutics on individual populations of cells, or even specific molecules. If these techniques prove effective in mice, they might be translated into the clinic in the future, where they could be used to non-invasively detect and monitor treatment of human cancers. PMID- 11902582 TI - The promise and peril of surrogate end points in cancer research. AB - Both experimental and observational studies of cancer need to have an end point. Traditionally, in aetiological and prevention studies, that end point has been the incidence of cancer itself, whereas in therapeutic trials, the end point is usually time to cancer recurrence or death. But cancer takes a long time to develop in an individual and is rare in the population. Therefore, aetiological studies and prevention trials must be large and lengthy to be meaningful. Similarly, many therapeutic trials require a long follow-up of large numbers of patients. Surrogate end points--markers of preclinical cancer or of imminent recurrence--are therefore an attractive alternative. But how can we be sure that a study with a surrogate outcome gives us the right answer about the true end point? PMID- 11902583 TI - Helicobacter pylori and gastrointestinal tract adenocarcinomas. AB - Although gastric adenocarcinoma is associated with the presence of Helicobacter pylori in the stomach, only a small fraction of colonized individuals develop this common malignancy. H. pylori strain and host genotypes probably influence the risk of carcinogenesis by differentially affecting host inflammatory responses and epithelial-cell physiology. Understanding the host-microbial interactions that lead to neoplasia will improve cancer-targeted therapeutics and diagnostics, and provide mechanistic insights into other malignancies that arise within the context of microbially initiated inflammatory states. PMID- 11902584 TI - Hypoxia--a key regulatory factor in tumour growth. AB - Cells undergo a variety of biological responses when placed in hypoxic conditions, including activation of signalling pathways that regulate proliferation, angiogenesis and death. Cancer cells have adapted these pathways, allowing tumours to survive and even grow under hypoxic conditions, and tumour hypoxia is associated with poor prognosis and resistance to radiation therapy. Many elements of the hypoxia-response pathway are therefore good candidates for therapeutic targeting. PMID- 11902585 TI - Multidrug resistance in cancer: role of ATP-dependent transporters. AB - Chemotherapeutics are the most effective treatment for metastatic tumours. However, the ability of cancer cells to become simultaneously resistant to different drugs--a trait known as multidrug resistance--remains a significant impediment to successful chemotherapy. Three decades of multidrug-resistance research have identified a myriad of ways in which cancer cells can elude chemotherapy, and it has become apparent that resistance exists against every effective drug, even our newest agents. Therefore, the ability to predict and circumvent drug resistance is likely to improve chemotherapy. PMID- 11902586 TI - Immune evasion in human papillomavirus-associated cervical cancer. AB - Tumour-associated viruses produce antigens that, on the face of it, are ideal targets for immunotherapy. Unfortunately, these viruses are experts at avoiding or subverting the host immune response. Cervical-cancer-associated human papillomavirus (HPV) has a battery of immune-evasion mechanisms at its disposal that could confound attempts at HPV-directed immunotherapy. Other virally associated human cancers might prove similarly refractive to immuno-intervention unless we learn how to circumvent their strategies for immune evasion. PMID- 11902587 TI - The case for population-based screening for colorectal cancer. AB - Screening for colorectal cancer is commanding increasing attention. Other cancer screening programmes have been a part of public consciousness for some time, but, until recently, colorectal cancer screening has remained in the background. Fuelled by new research, market opportunities and increased recognition of individual risk, screening for colorectal cancer is becoming a recommended procedure, but controversy about how best to implement widespread screening remains. PMID- 11902588 TI - Landmark papers in multiple sclerosis. PMID- 11902589 TI - Interferon beta-lb is effective in relapsing-remitting multiple sclerosis. II. MRI analysis results of a multicenter, randomized, double-blind, placebo controlled trial. 1993 [classical article]. PMID- 11902590 TI - Copolymer 1 reduces relapse rate and improves disability in relapsing-remitting multiple sclerosis: results of a phase III multicenter, double-blind, placebo controlled trial. 1995. PMID- 11902592 TI - Interferon beta-lb is effective in relapsing-remitting multiple sclerosis. I. Clinical results of a multicenter, randomized, double-blind, placebo-controlled trial. 1993 [classical article]. PMID- 11902591 TI - Impact of interferon beta-1a on neurologic disability in relapsing multiple sclerosis. 1997. PMID- 11902594 TI - The 5-year risk of MS after optic neuritis: experience of the optic neuritis treatment trial. 1997. PMID- 11902593 TI - The impact of blinding on the results of a randomized, placebo-controlled multiple sclerosis clinical trial. 1994 [classical article]. PMID- 11902595 TI - Extended use of glatiramer acetate (Copaxone) is well tolerated and maintains its clinical effect on multiple sclerosis relapse rate and degree of disability. 1998 [classical article]. PMID- 11902596 TI - Annotation: Chronic Fatigue Syndrome in children and adolescents. AB - BACKGROUND: Over the past two decades Chronic Fatigue Syndrome (CFS) of childhood has gained increasing prominence. A number of clinical reports and case-control studies have examined the nature of the disorder, its associations, response to treatment and outcome. METHOD: A review of publications on childhood CFS was undertaken and reference to work on adult CFS made. Most studies on childhood CFS have been on markedly affected children attending specialist pediatric clinics and very little is known about the condition as it presents in the community or to general medical services. RESULTS: The main symptom is fatigue in association with a variety of physical symptoms and with marked and prolonged functional impairment. CFS is commonly reported as being brought on by acute infections. Co morbid psychiatric (usually mood) disorders are present in at least a half. Personality problems and health attitudes have been described as possible predisposing and maintaining factors. Clinical reports indicate that family work focused on engagement and on a rehabilitation programme (including graded increasing activity and treatment of psychiatric co-morbidity) can help even the more severely impaired children. Recovery may be expected in over two-thirds. CONCLUSIONS: CFS presents as a distinct, markedly impairing disorder of childhood. In its severe form, it is often associated with mood disorders. Further research into milder forms and into the efficacy of different treatment interventions is specially needed. PMID- 11902597 TI - Deviant partner involvement and offending risk in early adulthood. AB - BACKGROUND: This paper examines the effects of deviant and non-deviant partner involvement at age 21 on patterns of continuity and change in offending between the ages of 18 and 21 years in a birth cohort of 983 young men and women. RESULTS: Results showed that those involved with a non-deviant partner had lower rates of offending at age 21 than those with no partner, whilst those without a partner had lower rates of offending at age 21 than those involved with a deviant partner. Associations between deviant/non-deviant partner involvement and offending risk best fitted a main effects model in which both offending at age 18 and young people's partnership choices at age 21 made independent and additive contributions to the prediction of offending at age 21. There was no interaction between offending at age 18 and partner choice at age 21 in determining offending risk in early adulthood. In addition, the effects of deviant/non-deviant partner involvement on patterns of offending were the same for men and women, and were found to persist even after extensive control for the confounding effects of a wide range of selection factors measured during childhood and adolescence. CONCLUSION: These results highlight the importance of partnership choices during early adulthood in determining young people's risk of offending as adults. PMID- 11902598 TI - Differences between preschool children with ODD, ADHD, and ODD+ADHD symptoms. AB - BACKGROUND: This study examines differences between children (ages 3 to 6 years) who have the symptoms of oppositional defiant disorder (ODD) with or without attention-deficit/hyperactivity disorder (ADHD), ADHD alone, and a nonODD/ADHD comparison group. Parent (N=595) and teacher (N=538) ratings were obtained for children attending the same community early childhood programs and for youngsters evaluated in an outpatient clinic (N=224) using a DSM-IV-referenced rating scale. RESULTS: Differences between symptom groups varied depending on how they were configured (teacher versus parent ratings) and setting (clinic versus community). In general, the ODD+ADHD group received the highest (and the comparison group the lowest) ratings of severity for the symptoms of other disorders, difficulties with peers, and developmental deficits. Moreover, the clinical impact of comorbidity was largely additive. Differences between youngsters with ODD versus ADHD symptoms were most apparent for teacher-defined groups in the community sample and parent-defined groups in the clinic sample. CONCLUSIONS: Collectively, these findings provide preliminary evidence for the notion that ODD and ADHD may constitute distinct clinical entities in preschool-aged children and suggest that informant may be an important consideration in the formulation of diagnostic criteria. PMID- 11902599 TI - Post-traumatic stress in children following motor vehicle accidents. AB - BACKGROUND: This study examined children's experiences following a motor vehicle accident (MVA). METHODS: Approximately 9 months following the accident, children (n=50) and their parents (n=50) participated in extensive interviews about the accident and in comprehensive, structured diagnostic interviews concerning overall psychological functioning. Additional assessments included post-traumatic stress questionnaires, archival police report records, and emergency treatment medical records. RESULTS: Of the 50 children, 7 children (14%) met criteria for PTSD diagnosis, and an additional 5 children met criteria for specific phobia (10%) related to the automobile accident on the structured diagnostic interview (DICA-R-C; total of 24%). Degree of physical injury predicted more PTSD symptoms, and previous accident experiences predicted fewer symptoms, before and after controlling for other variables. Holding degree of physical injury and age constant revealed that social support predicted fewer PTSD symptoms. CONCLUSIONS: Findings suggest the possible inoculating role of previous accidents and the importance of social support following MVA injury. PMID- 11902600 TI - Coping with birthparent loss in adopted children. AB - BACKGROUND: Relationships among adopted children's appraisals of birthparent loss, their coping strategies for managing such loss, and child and parent reports of child adjustment were investigated within the context of a stress and coping model of adoption adjustment. METHODS: Eighty-two 8-12-year-old adopted children and one of their parents participated. Children completed questionnaires assessing their negative affect about birthparent loss, their curiosity about birthparents, their use of coping strategies to manage birthparent-related distress, and their levels of depression, anxiety, and global self-worth. Parents reported on children's externalizing and internalizing behavior problems and social competence. RESULTS: Children who reported higher levels of negative affect about birthparent loss also reported higher levels of depression and lower self-worth. Curiosity about birthparents predicted parent-rated externalizing behavior. Behavioral avoidant coping was associated with greater self-reported anxiety and parent-rated externalizing behavior, whereas problem solving coping was associated with increased parent-rated social competence. CONCLUSIONS: The findings, though limited by issues of measurement and sampling, add to the knowledge base regarding adopted children's appraisal and coping behaviors, and provide partial support for a stress and coping model of adopted children's adjustment. PMID- 11902601 TI - Psychosocial adjustment in siblings of children with autism. AB - BACKGROUND: This study investigated psychosocial adjustment in siblings of children with autism compared to siblings of children with Down syndrome and siblings of normally developing children. In addition, the relationships between feelings of loneliness, social support and psychosocial adjustment, and the influence of gender and family size on psychological adjustment were examined. METHODS: Ninety siblings (30 per group) between the ages of 8 and 18 and one parent of each child participated in this study. RESULTS: Results indicated that siblings of children with autism, as well as comparison siblings, were well adjusted and reported low levels of loneliness. Siblings of children with autism also reported that they received high levels of social support in their lives. CONCLUSIONS: Large family size appears to facilitate healthy adjustment in siblings of children with autism. PMID- 11902602 TI - The separateness of social and emotional loneliness in childhood. AB - BACKGROUND: Much of the childhood loneliness research is misleading because it confounds objective and subjective measures of loneliness. The overall aim of this research was to examine the relationship between social isolation and emotional loneliness. METHOD: Three extreme groups were identified in a sample of 640 4-9-year-old children. There were two ('rejected' [N=60] and 'lonely' [N=146]) in which social and emotional loneliness were unrelated. The first were socially isolated (rejected) but they did not feel lonely. The second group felt lonely but they were not socially isolated. The third group ('rejected/ lonely') consisted of 61 children who were rejected and also felt lonely. RESULTS: Felt loneliness and social rejection were experienced together by 61 children, but 206 children experienced either one or the other, but not both. The fourth and largest group [N=374] were neither rejected nor lonely. Differences between the groups were found on direct observation measures of solitariness, sociability, and aggression; peer reports of shyness, aggression, prosocial behaviour, disruptive behaviour and inability to take teasing; self-reports of self-worth and competence, self-reports of supportive relationships; and measures of language use. CONCLUSIONS: The results indicate that it is loneliness and not rejection that co-occurs with emotional problems. PMID- 11902603 TI - Social communication in children with epilepsy. AB - BACKGROUND: This study examined measures of social communication that involve the use of language in formulating and organizing thoughts and its relationship with seizure-related, developmental, cognitive, and behavioral variables in 92 children with complex partial seizure disorder (CPS), 51 with primary generalized epilepsy (PGE), and 117 normal children, aged 5.1-16.9 years. METHODS: Coding the children's speech samples with the Kiddie Formal Thought Disorder Rating Scale (Caplan et al., 1989) and Halliday and Hasan's (1976) analysis of cohesion demonstrated social communication deficits in both seizure disorder groups. RESULTS: The CPS patients had both formal thought disorder and cohesion deficits and the PGE group had mild cohesion deficits. IQ, as well as fronto-temporal and bilateral spike and wave activity were associated with the severity of the social communication deficits of the CPS group. The social communication deficits of the PGE group, however, were related to IQ and seizure control. CONCLUSIONS: Recurrent CPS and PGE and fronto-temporal localization of epileptic activity might impair the development of children's communication skills. PMID- 11902604 TI - High motion coherence thresholds in children with autism. AB - BACKGROUND: We assessed motion processing in a group of high functioning children with autism and a group of typically developing children, using a coherent motion detection task. METHOD: Twenty-five children with autism (mean age 11 years, 8 months) and 22 typically developing children matched for non-verbal mental ability and chronological age were required to detect the direction of moving dots in a random dot kinematogram. RESULTS: The group of children with autism showed significantly higher motion coherence thresholds than the typically developing children (i.e., they showed an impaired ability to detect coherent motion). CONCLUSIONS: This finding suggests that some individuals with autism may show impairments in low-level visual processing--specifically in the magnocellular visual pathway. The findings are discussed in terms of implications for higher-level cognitive theories of autism, and the suggestion is made that more work needs to be carried out to further investigate low-level visual processing in autism. PMID- 11902605 TI - Frontal brain activation in anxious school children. AB - BACKGROUND: An atypical EEG pattern of frontal brain activation has been found in infants and adults with emotional disorders. Eighty-two 8-year-old children and 56 11-year-old children were examined with regard to the following questions: 1. Do children who are diagnosed with anxiety disorder exhibit an atypical pattern of frontal brain activation? 2. Can this pattern be demonstrated in children of different ages? and 3. Are there gender differences in these patterns similar to those that have been demonstrated in adults? Baseline EEG activity was subjected to power spectral analysis. RESULTS: In 8- and 11-year-old anxious girls, the well-known pattern of greater right than left frontal activation emerged that has been found previously in internalizing preschoolers. Healthy girls showed no frontal asymmetry at 8 years of age, and a greater left than right frontal brain activation at 11 years. In contrast, healthy boys demonstrated a significantly greater right than left frontal activation, whereas anxious boys displayed no frontal asymmetry at the age of 8, and a greater left than right frontal activation at the age of 11. CONCLUSIONS: Children suffering from anxiety disorders exhibited a significantly different pattern of frontal brain activation than healthy children without any lifetime diagnosis of mental disorder. Distinct gender differences in frontal activation were found in anxious as well as in healthy children. This could be demonstrated in two samples of different ages. PMID- 11902606 TI - Prosthodontic biomaterials and adverse reactions: a critical review of the clinical and research literature. AB - Prosthodontic biomaterials include impression materials, luting cements, and restorative materials. They consist of metals and alloys ceramics, and polymer materials and are retained in patients for <60 min or for decades. Oral release of compounds from biomaterials occurs, and adverse reactions may follow dental treatment. Especially in allergically vulnerable patients contact allergy may occur. There are reports from many different countries on contact allergy from gold/palladium alloys, components from polymer-based materials, chromium/cobalt alloys, and nickel. Notifications on adverse reactions in Norway, Sweden, and England are handled by a registry in which patient reactions and occupational exposure are recorded. Data from The Adverse Reaction Unit in Bergen and Umea have been a most valuable basis in extending knowledge in a field of current interest in dentistry. A review of the clinical and research literature relating to prosthodontic biomaterials and adverse reactions shows that reliable methods seem necessary to expose the frequency of adverse reactions in general dentistry, including prosthodontic treatment. PMID- 11902607 TI - Comparative analysis of some mouthrinses on the production of volatile sulfur containing compounds. AB - Oral malodor is mainly caused by the presence of volatile sulfur-containing compounds (VSC) produced by proteolytic periodontopathic bacteria in the oral cavity. Different solutions have been used as mouthrinses, trying to reduce malodor, and a large number is on the market. The aim of this study was to compare the effect of three commercially available mouthrinses with a simple inexpensive solution of zinc (zinc acetate 0.1%) on the production of VSC in vivo. Two of the solutions contained triclosan, one of them with fluoride and the other with sodium bicarbonate, and the third one contained herbal components. Seven healthy subjects rinsed with cysteine to induce production of VSC at baseline. After halitosis induction and VSC measurements, the subjects rinsed with the test solution, and mouth airVSC analyses were then performed by means of gas chromatography subsequent to repeated cysteine rinses after 30, 60, and 120 min. The data were calculated as percentage reduction of VSC from baseline. The percentage reduction of VSC decreased over time for all experimental groups. Zinc acetate had clearly the highest percentage reduction, starting from 95.68% at 30 min and with 69.27% after 2 h. The three other mouthrinses produced a VSC reduction of 23.92% 49.86% after 30 min, decreasing to 13.06%-37.09% after 2 h. One-way ANOVA (P = 0.05) was applied, and comparisons showed no differences between the commercially available solutions, but zinc acetate was significantly better than these. It may be concluded that some commercial mouthrinses are markedly less effective than a simple and cheap solution of zinc acetate. PMID- 11902608 TI - Dental care utilization: a study of 50- to 75-year-olds in southern Sweden. AB - This study investigates dental care utilization in an adult population in Southern Sweden in relation to dental and social conditions, attitudes to costs, and perceived need to obtain dental services. The study was based on responses to a questionnaire sent in 1998 to a random sample, 1974 persons, aged 56-75 years. The response rate was 66%. A significantly higher probability of dental care utilization less than once a year was found for men, for those with few remaining teeth, and for those with removable dentures. A higher probability of dental care utilization less than once a year was found for those who stated perceived need to obtain dental care with no possibility because of the cost and for those who stated that the cost had influenced their attendance for dental care. The results showed that there were differences for sex and dental conditions in dental care utilization and that dental care utilization was related to attitudes towards costs of dental care. PMID- 11902609 TI - Placement and replacement of restorations in primary teeth. AB - This practice-based study aimed to record the use of restorative materials, the type of restoration by class, and the reason for and the age of failed restorations in primary teeth by means of a survey of placement and replacement of restorations in 1996 and 2000/2001. Written alternative criteria for placement and replacement of restorations were provided for the participating clinicians. Details on 2281 restorations showed that primary caries was the main reason for inserting restorations in primary teeth. Replacements of failed restorations represented 14% of the fillings (n = 2040) in 1996 and 9% in 2000/2001 (n = 241). More than 80% or the fillings in primary teeth were of tooth-colored material, predominantly of the light-cured type. About 50% of failed amalgam and glass ionomer-type restorations were replaced due to secondary caries. The median age of amalgam restorations (3 years) was significantly higher than that of tooth colored restorations (2 years). Any possible advantage of a cariostatic effect of glass ionomer-type materials is apparently annulled by their short longevity compared with amalgam. PMID- 11902610 TI - Effect on prostaglandin E2 and leukotriene B4 levels by local administration of glucocorticoid in human masseter muscle myalgia. AB - Our aim was to determine whether masseter muscle (M) and plasma (P) levels of prostaglandin E2 (PGE2) or leukotriene B4 (LTB4) are influenced by local glucocorticoid administration and whether such changes would be associated with corresponding changes in local pain or hyperalgesia. Eighteen patients with fibromyalgia and 15 with local masseter myalgia were examined immediately before and 2 weeks after intramuscular administration of glucocorticoid with regard to masseter muscle resting pain and tenderness to palpation, pressure pain threshold, maximum voluntary mouth opening (MVM), and pain on maximum voluntary mouth opening. The primary criteria for inclusion were presence of pain for a period of at least 3 months and tenderness to digital palpation in the masseter muscle region. At both visits microdialysis samples were obtained from the masseter muscle, and venous blood was collected for analysis of PGE2 and LTB4. Dialysate levels of M-PGE2 did not change significantly after glucocorticoid administration, but reduction of masseter resting pain and increase of MVM were associated with decrease of M-PGE2 in the patients with fibromyalgia. Dialysate levels of M-LTB4 increased in both groups. In the patients with local myalgia the plasma level of LTB4 also increased, and this increase was associated with a decrease of pain and masseter tenderness. In conclusion, this study shows that reduction of masseter level of PGE2 after intramuscular glucocorticoid administration is associated with a decrease of resting pain in patients with fibromyalgia. In addition, the masseter muscle level of LTB4 increases in patients with fibromyalgia and local myalgia. PMID- 11902611 TI - Effect of a hydrophobic tooth coating on gingival health, mutans streptococci, and enamel demineralization in adolescents with fixed orthodontic appliances. AB - The effect of an anti-adhesive enamel coating on plaque accumulation, gingival health, and enamel demineralization was evaluated in 39 adolescents undergoing treatment with fixed orthodontic appliances using a prospective split-mouth design. Immediately after bracket insertion, the polymer was randomly applied after enamel etching to the buccal surfaces of the teeth in the left or right upper quadrant, leaving the opposite quadrant as an untreated control (218 test and 216 control teeth). Reapplications were carried out every 3rd month during the course or the study. The following data were collected at baseline and at designated follow-ups: visible plaque index, total viable counts and proportion of mutans streptococci in plaque samples, gingival bleeding index, and amount of gingival crevicular fluid. The incidence of enamel demineralization adjacent to the appliances was scored clinically at the termination of the orthodontic treatment. The observation time ranged from 6-24 months, during which the participants used fluoride rinses and toothpaste daily. A slightly impaired gingival health and increased levels or mutans streptococci compared with baseline was disclosed during the treatment. Five subjects showed enamel demineralization on a total of 30 teeth at the time of de-bonding. The results indicated no statistically significant differences between the enamel-coated and untreated teeth with regard to the studied variables at any follow-up. In conclusion, the present findings did not support a clinically beneficial effect of the polymeric tooth coating in a low-caries group of adolescents treated with fixed orthodontic appliances. PMID- 11902612 TI - Nursing personnel's views on oral health from a health promotion perspective: a grounded theory analysis. AB - The aim of this study was to develop a model for how nursing personnel view oral health in general and the oral health of the care receivers in particular, applying a health promotion perspective and using grounded theory analysis. Data were collected through interviews with 17 nursing personnel, selected by strategic sampling. Analysis of the transcribed interviews showed that there were four strategies, related to staff education, hospital resources, and leadership motivation. The strategies were grounded in data and emerged from the interaction between the two main categories: 'the valuation of the importance of oral health' and 'the behavior towards oral health maintenance'. They were characterized as the routine, theoretical, practical, and flexible strategies, with the latter considered ideal. As increased knowledge is one important part in enhancing the nursing personnel's ability to perform oral hygiene procedures, there is a need for education among nursing personnel, primarily among those using a routine strategy. PMID- 11902613 TI - Long-term physical inactivity and oral health in Finnish adults with intellectual disability. AB - Physical inactivity is prevalent among patients with intellectual disability. Because little is known about the oral effects of poor mobility, we reviewed the medical and dental charts of institutionalized dentate patients (n = 214; 40.2 years +/- 12.1) of the Special Welfare District of Southwestern Finland. The number of decayed, missing, and filled teeth (DMFT), the number of retained teeth, dental treatment visits, and the type of the first treatment visit were recorded. Physical activity was good in 55% and severely reduced or completely absent in 45% of the patients. The degree of intellectual disability was mild or moderate in 40% and severe or profound in 60% of the patients. The walking patients weighed more (64.3 (19.6) versus 44.4 (14.4) kg; P< 0.001), had fewer secondary diagnoses (1.4 (1.3) versus 2.2 (1.4); P< 0.001), fewer daily medications (4.0 (2.1) versus 4.8 (2.4); P< 0.02), higher DMFT scores (18.5 (8.2) versus 14.8 (9.2); P < 0.05), and more dental treatment visits (2.7 (2.4) versus 2.0 (1.3); P< 0.03) than patients with poor physical activity. Periodontal treatment given as the primary type of dental care was more common among subjects with poor mobility than among those with good motor activity (P < 0.002). Poor physical activity was related to better dental health, higher need for periodontal therapy, and fewer dental visits than in patients with good motor activity. PMID- 11902614 TI - Prevalence of short-root anomaly in healthy young adults. AB - Short-root anomaly (SRA), occurring mostly in maxillary incisors, is defined as developmentally very short, blunt dental roots. The condition has a genetic background and is related to hypodontia. Earlier population studies have been based on schoolchildren with developing dentitions and have indicated prevalence figures between 1% and 10%. We studied a random sample of existing panoramic radiographs of 2000 university students for SRA. Roots as long as or shorter than the crowns in the incisors and visually evaluated as very short, blunt roots bilaterally in the posterior teeth were classified as SRA. The prevalence was 1.3%. According to anamnestic information, half the SRA patients had undergone orthodontic therapy, but pre-treatment radiographs were unavailable. In 70% of the SRA patients the short-rooted tooth pairs were upper incisors, but also involved were maxillary premolars, lateral incisors, and lower second premolars. Women were significantly more often affected. We discuss other factors known to cause short-rooted teeth and conclude that the population prevalence for genetic SRA in fully developed dentitions is close to our 1.3%. PMID- 11902615 TI - Wet or dry, normal or deproteinized dentin surfaces as substrate for dentin adhesives. AB - Bond strength between resin composite and dentin mediated by several dentin adhesives applied to dry or wet acid-etched dentin or to dry or wet acid-etched and deproteinized dentin were measured and analyzed. Human dentin were A) acid etched and blot-dried for 1 sec (= wet), B) as A but dried with air for 10 sec (= dry), C) acid-etched and treated with hypochlorite and then dried for 1 sec, and D) as C but dried with air for 10 sec. Eight dentin adhesives were used in each group for bond strength measurements. The results were compared by means of ANOVA and the Tukey test. Collagen removal increased significantly the strength of the bond by 10-18 MPa for five adhesives when tested dry (D versus A). When tested wet, collagen removal increased the strength by 10-12 MPa for three adhesives (C versus A). Normal etched dentin showed a reduction in strength of 14-15 MPa for three or the adhesives when tested dry instead of wet (B versus A). For one dentin adhesive no significant change in bond strength due to collagen removal and/or drying conditions was observed. It was hypothesized that low technique sensitivity of an adhesive may be linked to its ability to wet and adhere to collapsed collagen fibers and to the surface of the underlying mineralized tissue. Comparisons of bond strengths obtained by using dried or wet acid-etched dentin and dried or wet acid-etched and deproteinized dentin may be useful for evaluating the efficiencies or dentin adhesives. PMID- 11902616 TI - Perceiving personal discrimination: the role of group status and legitimizing ideology. AB - It was hypothesized that relative group status and endorsement of ideologies that legitimize group status differences moderate attributions to discrimination in intergroup encounters. According to the status-legitimacy hypothesis, the more members of low-status groups endorse the ideology of individual mobility, the less likely they are to attribute negative outcomes from higher status group members to discrimination. In contrast, the more members of high-status groups endorse individual mobility, the more likely they are to attribute negative outcomes from lower status group members to discrimination. Results from 3 studies using 2 different methodologies provide support for this hypothesis among members of different high-status (European Americans and men) and low-status (African Americans, Latino Americans, and women) groups. PMID- 11902617 TI - The dynamic time course of stereotype activation: activation, dissipation, and resurrection. AB - Stereotypes activated upon initial exposure to a stereotyped individual may dissipate as the exposure continues. Participants observing a videotaped interview with a Black person showed activation of the stereotype of Black people following 15 s of observation but not following 12 min of observation. However, the discovery of a disagreement with the stereotyped individual may bring the dissipated stereotype back to mind. Participants who discovered, at the end of a 12-min videotaped interview with a Black person, that this person disagreed with them about the verdict in a court case showed activation of the stereotype of Black people, whereas participants who discovered instead that the Black person agreed with them did not. Participants who disagreed with a Black person also applied the Black stereotype to him, but this stereotype application was detected only on an implicit measure of application, not on an explicit measure. PMID- 11902618 TI - Subtyping versus bookkeeping in stereotype learning and change: connectionist simulations and empirical findings. AB - A distributed connectionist network can account for both bookkeeping (M. Rothbart, 1981) and subtyping (M. B. Brewer, V. Dull, & L. Lui, 1981; S. E. Taylor, 1981) effects. The finding traditionally regarded as demonstrating subtyping is that exposure to moderate (compared with extreme) disconfirmers leads to subsequent ratings of the group that are less stereotypic. Despite learning that is incremental and analogous to bookkeeping, the simulations replicate this finding and suggest that the "subtyping" pattern of results will be drastically reduced if disconfirmers are encountered before the stereotype is well-established. This novel prediction holds with human participants and offers a tantalizing suggestion: Although moderate disconfirmers may produce more stereotype change. stereotype development might be discouraged by exposure to either extreme or moderate disconfirmers. PMID- 11902619 TI - The inaction effect in the psychology of regret. AB - Previous research showed that decisions to act (i.e., actions) produce more regret than decisions not to act (i.e., inactions). This previous research focused on decisions made in isolation and ignored that decisions are often made in response to earlier outcomes. The authors show in 4 experiments that these prior outcomes may promote action and hence make inaction more abnormal. They manipulated information about a prior outcome. As hypothesized, when prior outcomes were positive or absent, people attributed more regret to action than to inaction. However, as predicted and counter to previous research, following negative prior outcomes, more regret was attributed to inaction, a finding that the authors label the inaction effect. Experiment 4, showing differential effects for regret and disappointment, demonstrates the need for emotion-specific predictions. PMID- 11902621 TI - Could it happen to you? Predicting the impact of downward comparisons on the self. AB - Three studies examined the impact of downward comparisons on the self. Worse-off others exerted an impact only when participants drew an analogy between themselves and the other. When participants did draw an analogy, the impact of the other on the self was determined by perceived vulnerability to the other's negative fate. When vulnerability was low, downward comparisons enhanced self evaluations. When vulnerability was high, downward comparisons deflated self evaluations, but activated a prevention orientation, boosting motivation aimed at avoiding the negative experience of the other. PMID- 11902620 TI - The spontaneous use of a group typology as an organizing principle in memory. AB - Five studies investigated the spontaneous use of group typology in encoding information about various social groups. Participants saw faces or behaviors along with a label indicating the group membership of the face or the behavior. Labels corresponded to 2 groups each of 3 group types (i.e., 2 intimacy groups, 2 task-oriented groups, and 2 social categories). Recognition results showed more within-group-type errors than between-group-types errors. A free-recall task replicated these results, as the sequence of remembering items showed that memory organization reflected the group typology. A final study investigated the effects of group typology on the speed and accuracy of category membership verification. Results demonstrate the spontaneous use of an implicit group typology and its influence on the cognitive organization of information about groups. PMID- 11902622 TI - Social norms and the expression and suppression of prejudice: the struggle for internalization. AB - The authors studied social norms and prejudice using M. Sherif and C. W. Sherif's (1953) group norm theory of attitudes. In 7 studies (N = 1,504), social norms were measured and manipulated to examine their effects on prejudice; both normatively proscribed and normatively prescribed forms of prejudice were included. The public expression of prejudice toward 105 social groups was very highly correlated with social approval of that expression. Participants closely adhere to social norms when expressing prejudice, evaluating scenarios of discrimination, and reacting to hostile jokes. The authors reconceptualized the source of motivation to suppress prejudice in terms of identifying with new reference groups and adapting oneself to fit new norms. Suppression scales seem to measure patterns of concern about group norms rather than personal commitments to reducing prejudice; high suppressors are strong norm followers. Compared with low suppressors, high suppressors follow normative rules more closely and are more strongly influenced by shifts in local social norms. There is much value in continuing the study of normative influence and self-adaptation to social norms, particularly in terms of the group norm theory of attitudes. PMID- 11902623 TI - A room with a cue: personality judgments based on offices and bedrooms. AB - The authors articulate a model specifying links between (a) individuals and the physical environments they occupy and (b) the environments and observers' impressions of the occupants. Two studies examined the basic phenomena underlying this model: Interobserver consensus, observer accuracy, cue utilization, and cue validity. Observer ratings based purely on offices or bedrooms were compared with self- and peer ratings of occupants and with physical features of the environments. Findings, which varied slightly across contexts and traits, suggest that (a) personal environments elicit similar impressions from independent observers, (b) observer impressions show some accuracy, (c) observers rely on valid cues in the rooms to form impressions of occupants, and (d) sex and race stereotypes partially mediate observer consensus and accuracy. Consensus and accuracy correlations were generally stronger than those found in zero acquaintance research. PMID- 11902624 TI - Thinking about oneself and others: the relational-interdependent self-construal and social cognition. AB - These studies focus on the relational-interdependent self-construal's association with implicit or indirect cognitive processes. In the relational-interdependent self-construal, the self is defined largely in terms of close relationships, resulting in variation in self-related processes. In Studies 1 and 2, the relational self-construal was associated with positive implicit evaluations of relational concepts and with tightly organized cognitive networks of relational terms. Studies 3 and 4 demonstrated that this self-construal was associated with memory for and implicit organization of relational information. In Studies 5 and 6, the relational self-construal was positively related to the degree to which participants described themselves and a friend similarly. The implications of the relational self-construal for theories of relationship cognition and for other self-related cognitive processes are discussed. PMID- 11902625 TI - Adult attachment and the perception of facial expression of emotion. AB - Adult attachment orientation has been associated with specific patterns of emotion regulation. The present research examined the effects of attachment orientation on the perceptual processing of emotional stimuli. Experimental participants played computerized movies of faces that expressed happiness, sadness, and anger. Over the course of the movies, the facial expressions became neutral. Participants reported the frame at which the initial expression no longer appeared on the face. Under conditions of no distress (Study 1), fearfully attached individuals saw the offset of both happiness and anger earlier, and preoccupied and dismissive individuals later, than the securely attached individuals. Under conditions of distress (Study 2), insecurely attached individuals perceived the offset of negative facial expressions as occurring later than did the secure individuals, and fearfully attached individuals saw the offset later than either of the other insecure groups. The mechanisms underlying the effects are considered. PMID- 11902626 TI - Motivated decision making: effects of activation and self-centrality of values on choices and behavior. AB - Six studies examined the value-behavior relation and focused on motivational properties of values, the self, and value activation. Priming environmental values enhanced attention to and the weight of information related to those values, which resulted in environmentally friendly consumer choices. This only occurred if these values were central to the self-concept. Value-congruent choices were also found in response to countervalue behavior in an unrelated context. Donating behavior congruent with central altruistic values was found as a result of enhanced self-focus, thus demonstrating the importance of the self in the value-behavior relation. The external validity of the value-centrality measure and its distinction from attitudes were demonstrated in the prediction of voting. Values were thus found to give meaning to, energize, and regulate value congruent behavior, but only if values were cognitively activated and central to the self. PMID- 11902627 TI - Measuring needs with the thematic apperception test: a psychometric study. AB - Three apperception theories that explain how people respond to Thematic Apperception Test cards are proposed: a simple apperception theory, an apperception theory with a dynamic component, and an apperception theory with 2 types of responses. Each theory is translated into an item response theory model and is applied to need for achievement (nAch) data. The analysis indicates that the best fitting model is provided by the apperception theory with 2 types of responses, also referred to as the drop-out apperception theory. The 1st type of response predicted by this theory is determined by the nAch level of the person and the achievement-response-eliciting value of the card; this response is diagnostic for the nAch level of the person. The 2nd type of response is not determined by the 2 aforementioned characteristics and is therefore not diagnostic of the person's nAch level. The results are cross-validated for need for power and need for affiliation. PMID- 11902628 TI - Treatment of patients with acute renal failure during Marmara earthquake. AB - Crush syndrome or traumatic rhabdomyolysis is a serious and epidemic clinical case that develops among casualties of earthquakes or other catastrophic events. Hypocalcaemia is one of the main symptoms that accompanies crush syndrome. It is essential that immediate IV volume replacement treatment be initiated for such patients, which is known to positively affect prognosis and help prevent the development of acute renal failure. Following the earthquake of 17th August 1999 in the north-west of Turkey, 333 casualties were hospitalised in the Medical Faculty of Uludag University. 60 (18%) of these patients [34 males (56.6%) and 26 females (43.4%)] received haemodialysis treatment. The average number of HD sessions received bythe patients was 48. Intermittent HD applied to 44 patients (73.3%) was the preferred method of treatment. Fluid overload and azotemia were the leading indications (76.6%) for dialysis. The lower extremities were mainly injured in these patients [33 (55%)]. Therefore, subclavian catheterisation was preferred in those patients with trauma at this site. 594 (67.1%) HD sessions were applied with limited heparin due to the risk of bleeding. Hypotension in 94 patients (61%) and thrombosis in 60 (45.7%) were the main complications. Of the complications not due to HD, infection in 23 patients (38.3%) was the most significant. 21 patients (35%) were lost during the treatment. It was thanks to the sacrifices and co-operation among national and international medical teams that the mortality rate was low. PMID- 11902629 TI - The social climate in chronic haemodialysis units as perceived by patients and nurses. AB - The purpose of this research was to check if there was a correlation between how patients perceived the atmosphere in dialysis units compared to the nurses' perception. The intention was to examine commonality and differences between the two groups. This would serve as a basis to change the behaviour of the nurses and/or to develop or create settings that are more suitable. Sixteen haemodialysis units in Israel participated in this study, a total of 190 participants of which ninety-three were nurses and ninety-seven were patients. Patients and nurses gave their consent in writing and answered anonymously. The Moos Ward Atmosphere Scale was used using 100 true and false questions. These questions were condensed into six main categories. Our results showed that the patients and staff had significantly different perceptions in the following categories: 1. Openness and sensitivity, 2. Staff attitude, 3. Order and organization, 4. Mutual support, 5. New treatment approaches. The greatest degree of agreement between the two groups was found in only one category: that of the "doctor's attitude" or behaviour. Three studies have been found that have investigated the unit atmosphere as perceived by patients and staff-two in Israel in an oncology and a psychiatric unit. Rhodes did the third study in a haemodialysis unit in the Unites States and his results are compared with the results in this study. The differences found between nurses and patients show that there is a communication problem. It is recommended that interpersonal communication be improved to close the gap in perceptions, thereby improving the unit atmosphere. New strategies should be developed for coping and helping the patient to adjust. PMID- 11902630 TI - Job-related stress among nursing personnel in Greek dialysis units. AB - The purpose of this study was to describe specific work-related factors that contribute to increased levels of stress experienced by nursing personnel, and to compare their impact on nurses and assistant nurses who work in Dialysis Units (DU) to those who work in Peritoneal Dialysis Units (PDU) in Greece. The sample of the study consisted of 682 members of nursing personnel working in DU and in PDU in Greek Hospitals. The collection of data was done by means of a questionnaire. The latter included questions about the motivation of nursing personnel for having chosen to work in these units along with questions about the stress factors related to the patients and their care, the role of the nurse in the unit, and the working conditions. According to the results of the study, the percentage of nursing personnel working in DU or PDU by their own choice was 71% and 8.3% respectively, whereas the rest of the staff were placed there irrespective of their preferences, by the Administrators. Among nursing personnel who had chosen to work in DU, the reported motives that contributed to their decision were the absence of a night shift (27.8%), and working in a closed unit. The main motive that incited nursing personnel to work in PDU was the acquisition and the application of specialized knowledge. The percentage of nursing personnel working in DU that expected high levels of job satisfaction was 77%, whereas the corresponding percentage for nursing personnel working in PDU was 65%. However, only 44% and 37% of the nursing personnel working in DU and PDU reported high levels of job satisfaction. The most important stressors related: i) To the patient: were the risk of contamination (79% DU, 84% PDU) and the death of a patient (77% DU, 80% PDU). 2) To the role of nursing personnel in the unit: were increased responsibilities (65% DU, 37% PDU), low involvement of the nursing personnel in decision making (58% DU, 54% PDU), and low professional status of nursing personnel. 3) To the working conditions: were the shortage of nursing personnel (74% DU, 99% PDU), limited material (74% DU, 57% PDU), the closed environment (75% DU, 64% PDU) and the daily work routine (78% DU, 61% PDU). In conclusion, we can say that working in DU and PDU provokes increased stress in nursing personnel, even though the implicated stress factors differ between these units. PMID- 11902631 TI - The relationship between interdialytic weight gain and patient compliance. A single centred cohort study (n=21). AB - Haemodialysis patients are subject to many restrictions and face a lifetime attempting to adhere to various regimes. This small study was designed to explore the relationship between fluid gain, compliance and staff reinforcement. This was achieved by measuring dietetic and nursing educational input using interdialytic weight gain as a barometer of patient compliance. This study suggests renal nurses, dietitians and other members of the multi-disciplinary team may have less influence than previously assumed. PMID- 11902632 TI - The grown-up patient. The new customer in dialysis or--how to handle the demanding and emancipated dialysis patient. AB - The treatment of dialysis patients is under pressure. As a result of strict budgeting and increased administrative work, enhancement and the further development of the dialysis health care system is needed. An essential element of that development is a radical change in the patient/nurse relationship. Customer relationship management assumes that the patient is seen as a client, is encouraged to make decisions on their treatment and also emphasises the professionalism of nursing. PMID- 11902633 TI - Subjective global assessment of nutrition a useful diagnostic tool for nurses? AB - Malnutrition in maintenance haemodialysis (HD) patients is closely related with morbidity and mortality in this population. AIM OF THE STUDY: To evaluate feasibility, performance and information given by SGA (subjective global assessment), a semi-quantitative method of nutritional evaluation (based on a medical questionnaire and a simple clinical examination) carried out by a group of 12 nurses. METHOD: In March 1999, a feasibility study was organised to evaluate 9 patients during HD. Since July 1999, a nutritional evaluation of all the patients meeting the required parameters (e.g. duration of HD > 6 months, day sessions) has been set up every 4 months. RESULTS: In March 1999, preliminary results demonstrated an average learning time per SGA of 15'. In July 1999, 32 patients were evaluated, albuminemia (micromol/l) and pre-albuminemia (g/l) were analysed according to the SGA classification (A = good nutrition, B= light to moderate malnutrition, C= severe malnutrition). The albuminemia and the prealbuminemia of patients A (respectively 541+/-45 and 0.37+/-0.10) were higher than those of patients B (482+/-41 and 0.31+/-0.01), and those of patients C (381+/-54 and 0.19+/-0.1), by variance analysis (p < 0.0001). The information drawn from the SGA reveal a severe malnutrition, a light to moderate malnutrition, and good nutrition in respectively 13%, 63% and 25% of the patients, and a noticeable muscular atrophy (moderate to severe) in 43% of cases. Anorexia and major gastro-intestinal symptom (nausea, vomiting, and diarrhoea) are found in 14% of cases. CONCLUSION: Beside traditional methods of screening and evaluation of malnutrition in HD patients, the use of SGA by a nursing team uncovers useful information on nutritional status of patients, especially in areas lacking facilities such as a laboratory, dietetic department or permanent presence of doctors. PMID- 11902634 TI - Nutritional screening tools for CAPD patients: are computers the way forward? AB - The aim of this study was to develop a computer programme (CP) to identify CAPD patients at risk of malnutrition as well as screen monthly biochemical results of transplant (tx), haemodialysis (HD), CAPD and nephrology (Ne) patients for abnormal levels such as hyperkalaemia. METHODS: The CP was designed using the programme Proton. Proton can automatically download biochemical results from pathology making it possible to generate a list of patients with results outside a desired biochemical range in accordance with national renal standards for adult patients. Biochemical measures of nutritional status were used to define malnutrition as 2 or more results outside of these parameters (see Table 1): 10% weight loss, Kt/V < 1.9, normalised protein catabolic rate (nPCR) < 1.2: predialysis urea < 15 mmol/L, phosphate (PO4) < 0.75 mmol/L, potassium (K), < 3.3 mmol/L. The CP was compared with standardised dietetic assessments (SDA) for validity. An SDA includes a review of medical, social history, biochemistry (blood and urine), dialysis prescriptions, treatment plans, medications, weight changes, BMI, dietary intake compared to therapeutic guidelines with documented aims. The therapeutic guidelines used to complete dietary assessment include: protein 1.2-1.5 g/kg Ideal Body Weight (IBW), energy: aim for BMI of 20-25 kg/m2, sodium: 100-120 mmol/L, K: 1 mmol/kg IBW, fluid: urine output + medical advice, PO4: 175-200 mg of PO4/10 g of protein. 3005 patients from Richard Bright Renal Unit in Bristol, England, including 4 Satellite HD units, had blood results screened for abnormal levels using CP (see Table 1). In the clinic setting, the length of time taken to review biochemical results was measured before and after implementation. RESULTS: The CP identified 88% of CAPD patients at risk of malnutrition compared with 67% by SDA. The time taken to look up biochemical results of patients attending the outpatient clinics was reduced by approximately 30 minutes leaving only the diet history computer screen to review for each patient. DISCUSSION: The CP should be compared with subjective global assessment (SGA) for validity in the identification of malnutrition. Serum cholesterol, prealbumin, protein equivalent of total nitrogen appearance (nPNA), C reactive protein (CRP) and bicarbonate should be incorporated into the CP in order to improve its specificity. CONCLUSION: Using the present parameters, the CP over diagnosed the number of CAPD patients at risk of malnutrition. However, the CP improves time management and rationalisation of dietetic activity by screening abnormal biochemistry. PMID- 11902636 TI - Education of patients and use of insulin pumps in diabetic treatment. AB - Diabetes Mellitus is a chronic disease well known for its fatal complications. Therefore, an international call for measures was declared to keep complications of diabetes to a minimum in order to reduce the costs of treating these complications (St. Vincent Declaration). The common international guidelines for treatment of patients with Type 1 Diabetes Mellitus are the Diabetes Control and Complications Trial (DCCT). The United Kingdom Prospective Diabetes Study (UKPDS) investigated the use of tight glycaemic control in those with Type 2 Diabetes Mellitus. This paper will explain the importance of intensive insulin treatment for diabetic patients, even when they are dialysing, to prevent diabetic complications. There's always a possibility of reducing diabetic complications to ensure a better quality of life. It will also discuss the different treatment options available including Continuous Subcutaneous Insulin Infusion, which has been demonstrated as the most effective mode of treatment. PMID- 11902635 TI - The nurses knowledge, awareness and acceptance of tissue-organ donation. AB - BACKGROUND: Organ donation and transplantation, although widely accepted as a successful medical procedure, is one of the new concepts in nursing. Organ donation does not occur as often as needed and the reasons for acceptance or refusal are not clear. To meet this demand more organs and tissues need to be recovered from potential donors. Nurses working on transplantation units are given in-service training and gain knowledge through experience. Nurses are in a position to inform, and to ask families to donate organs, and also to inform potential donors on their ward to the units. METHODS: A prospective, descriptive and semi-analytic study was designed. Data were collected using 25 structured and semi-structured questionnaires, over a week in April 2000. 3 general and 2 midwifery hospitals were visited and the purpose of the study explained. Staff were asked to participate assuring anonymity. A conference on the issue was offered on completion of the questionnaire if desired. The questionnaire came with an explanation of the purpose of the study, and thanked staff for their participation. They were given to the nurses in charge on each ward, and collected 3 days later by the researchers. The respond rate was 65.6%. RESULTS: Of the nurses, 87.7% had positive thoughts about the organ donation, but only 10.8% knew the donation law, 68.8% would consider donating organs of their own, 58.7% would consider signing a consent card, and only 36.7% would donate organs from their family members. Although, the majority had positive views about the issue only 34.4% showed willingness to talk to families and ask for donations, 84.0% would inform potential donors in the unit. Adequate knowledge and level of education were the factors effecting acceptance and willingness to be involved in organ harvesting efforts (p < 0.001 and p < 0.05). CONCLUSIONS: The only significant predictors of their acceptance and willingness were education (chi2 = 6.45, p < 0.05), and to have adequate knowledge (chi2 = 21.90, p < 0.001). Nurses were found in need of education about all aspects of brain death and organ donation including how and when to approach families to inform and ask for organs, and how to support families throughout the process. A brochure should be prepared in detail to guide them on this task. More research should be done to clarify the reasons for refusal of organ donation. PMID- 11902637 TI - CDC recommendations. PMID- 11902638 TI - Nursing classifications (NIC and NOC) and hepatitis C in dialysis. PMID- 11902639 TI - Latex allergy; a misjudged problem! PMID- 11902640 TI - Quinoxaline chemistry. Part 14. 4-(2-Quinoxalylamino)-phenylacetates and 4-(2 quinoxalylamino)-phenylacetyl-L-glutamates as analogues--homologues of classical antifolate agents. Synthesis and evaluation of in vitro anticancer activity. AB - Among a new series of 26 4-(3-substituted-2-quinoxalylamino)phenylacetates and 4 (3-substituted-2-quinoxalylamino)-phenylacetyl-L-glutamates, eight were selected at NCI for evaluation of their in vitro anticancer activity. The results obtained in comparison with the corresponding nor-compounds series seem to indicate that this type of homologation is not helpful. PMID- 11902641 TI - Synthesis and evaluation for biological activity of 3-alkyl and 3-halogenoalkyl quinoxalin-2-ones variously substituted. Part 4. AB - A new series of 3-isopropyl-, 3-trifluoromethyl- and 3-bromomethylquinoxaline-2 ones variously substituted on the benzo-moiety were synthesized and submitted to a preliminary in vitro evaluation for antibacterial, antifungal and anti-HIV activities. Furthermore, all compounds were also tested for cytotoxicity. Results of the screening showed that compound 10 exhibits moderate antimicrobial activity against Staphylococcus aureus (MIC = 33 microM), and that 25 and 26 showed interesting cytotoxicity versus mock-infected MT-4 cells. All the other compounds were inactive. PMID- 11902642 TI - Synthesis and biological activity of some 4-substituted 1-[1-(2,3-dihydroxy-1 propoxy)methyl-1,2,3-triazol-(4 & 5)-ylmethyl]-1H-pyrazolo[3,4-d]pyrimidines. AB - Cycloaddition of 7a, b with 6 gave, after separation and deprotection, two regioisomers 10a, b and 11a, b. The deprotected acyclic nucleoside 10a used as the precursor for the preparation of 4-amino (12), 4-methylamino (13), 4 benzylamino (14), 4-methoxy (15) and 4-hydroxy (16) analogues. All acyclic nucleosides were evaluated for their inhibitory effects against HIV-1(IIIB), HIV 2(ROD) in MT-4 cells, for their anti-tumor activity and for their inhibitory effects against Mycobacterium tuberculosis. No marked activity was found. PMID- 11902643 TI - Two new spectrophotometric approaches to the multicomponent analysis of the acetaminophen and caffeine in tablets by classical least-squares and principal component regression techniques. AB - Classical least-squares (CLS) and principal component regression (PCR) techniques were proposed for the simultaneous analysis of tablets containing acetaminophen and caffeine without using a chemical separation procedure. The chemometric calibrations were prepared by measuring the absorbances values at the 15 wavelengths in the spectral region 215-285 nm and by using a training set of the mixtures of both drugs in 0.1 M HCI. The obtained chemometric calibrations were used for the estimation of acetaminophen and caffeine in samples. The numerical calculations were performed with the 'MAPLE V' software. By applying two techniques to synthetic mixtures, the mean recoveries and the relative standard deviations in the CLS and PCR techniques were found as 99.5 and 1.29, 99.7 and 1.00% for acetaminophen and 99.9 and 1.92, 100.0 and 1.178% for caffeine, respectively. Our results were compared with those obtained previously by one of us considering HPLC method as a reference method. These two methods were successfully applied to a pharmaceutical tablet formulation of two drugs. PMID- 11902644 TI - Antimicrobial activity of 5-arylidene aromatic derivatives of hydantoin. Part 2. AB - Various 5-chloroarylidene-2-amino substituted derivatives of imidazoline-4-one were synthesized and evaluated for their activity in vitro against Mycobacterium tuberculosis and other type strains of bacteria and fungi. 2-Chloro- and 2,4 dichlorobenzylidene substituted hydantoins exhibited antimycobacterial effect. The most potent compounds 3i, 3j, 3o, 3q and 3s were classified for further tests. The antimitotic effect of the investigated hydantoins was also examined. PMID- 11902645 TI - Synthesis of (+)- and (-)-cis-2-[(1-adamantylamino)-methyl]-1-phenylcyclopropane derivatives as high affinity probes for sigma1 and sigma2 binding sites. AB - Selective ligands for either sigma1 (sigma1) or sigma2 binding sites are potentially useful for gaining a better understanding of the physiological functions of these proteins. Moreover, potent and selective homochiral sigma1 and sigma2 binding site ligands represent leads to potential radioligands for tumour imaging with positron emission tomography (PET). On the basis of their structural similarity to previous leads, new (+)- and (-)-cis-2-[(1-adamantylamino)-methyl] 1-phenylcyclopropane derivatives were synthesised and their binding affinities for sigma1 and sigma2 binding sites were determined. Each enantiomer showed high affinity for both sigma1 and sigma2 binding sites, but only (-)-cis-methyl-2-[[1 adamantyl(methyl)amino]methyl]-1-phenylcyclopropane-carboxylate, (-)-4, showed appreciable selectivity for binding to sigma1 versus sigma2 sites. The enantiomers of cis-(2-[[1-adamantyl(methyl)amino]methyl]-1 phenylcyclopropyl)methanol, 6, expressed the highest affinity for sigma1 and sigma2 binding sites. Ligands (-)-4, (+)-6 and (-)-6 might be rapidly labelled in their N-methyl groups by methylation of the N-desmethyl analogues with [11C]iodomethane to provide prospective radioligands for PET. The N-desmethyl analogues, which are also high affinity ligands, were prepared and shown to undergo satisfactory methylation with iodomethane. PMID- 11902647 TI - Synthesis and antibacterial activity of pyridazino[4,3-b]indole-4-carboxylic acids carrying different substituents at N-2. AB - The synthesis and the in vitro evaluation of antibacterial activity of new pyridazino[4,3-b]indole-4-carboxylic acids 2-4, 6 against some selected representative of Gram-positive and Gram-negative bacteria are reported. The role of the lipophilicity in the modulation of the antibacterial activity of the tested compounds is discussed. All the synthesized compounds appear quite weak against Gram-positive bacteria, whereas have no significant activity against Gram negative bacteria. Only derivative 2g possesses an interesting activity against Gram-positive bacteria. PMID- 11902646 TI - Influence of irradiation sterilization on poly(lactide-co-glycolide) microspheres containing anti-inflammatory drugs. AB - Gamma-irradiation is finding increasing use in the sterilization of pharmaceutical products. However, irradiation might also affect the performance of drug delivery systems. In this study, the influence of gamma-irradiation on the physicochemical properties of two commonly used non-steroidal anti inflammatory drugs (NSAIDs) [naproxen sodium (NS) and diclofenac sodium (DS)] was investigated. The drugs were incorporated in poly(lactide-co-glycolide) (PLGA, 50:50; molecular weight 34000 or 88000 Da) microspheres. The biodegradable microspheres were irradiated at doses of 5, 15, 25 kGy using a 60Co source. Drug loading of irradiated and non-irradiated microspheres with both 34000 and 88000 Da polymers were essentially the same. A significant difference was noticed in the particle sizes of the irradiated as compared to the non-irradiated formulations. Notably, in release studies, the amount of active substance released from PLGA microspheres showed an increase with increasing irradiation dose. In DSC, the glass transition temperatures (Tg) of microspheres exhibited a slow increase with irradiation dose. PMID- 11902648 TI - Synthesis and biological activity of 5-alkyl-6-(alkylsulfanyl)- or 5-alkyl-6 (arylsulfanyl)pyrazine-2-carboxamides and corresponding thioamides. AB - Nucleophilic substitution of chlorine in 5-alkyl-6-chloropyrazine-2-carboxamides with various alkyl and arylthiolates afforded 20 5-alkyl-6-(alkylsulfanyl)- and 5 alkyl-6-(arylsulfanyl)pyrazine-2-carboxamides. The reaction of the amides with Lawesson's reagent yielded the corresponding thioamides. The assessment of in vitro antimycobacterial and antifungal activity of the compounds was carried out. In both series, the antimycobacterial activity increases with increasing molecular weight of the alkylsulfanyl group in position 6 of the pyrazine ring. Thioamides exhibited higher activity than the corresponding amides. 5-Butyl-6 (phenylsulfanyl)pyrazine-2-carbothioamide (2j) possessed the highest activity (91% inhibition) against Mycobacterium tuberculosis and also the highest lipophilicity (log P = 4.95). Only a poor in vitro antifungal effect was noted in 5-butyl-6-(butylsulfanyl)pyrazine-2-carboxamide (1i) and 6-(ethylsulfanyl)-5 isobutylpyrazine-2-carbothioamide (2q) against Trichophyton mentagrophytes and Absidia corymbifera. PMID- 11902649 TI - Synthesis and antimycobacterial activity of 3-aryl-, 3-cyclohexyl- and 3 heteroaryl- substituted-2-(1H(2H)-benzotriazol-1(2)-yl)prop-2-enenitriles, prop-2 enamides and propenoic acids. II. AB - A series of 32 3-aryl-, 3-cyclohexyl-, and 3-heteroaryl-substituted-2-(1H(2H) benzotriazol-1(2)-yl)-prop-2-enenitriles, prop-2-enamides and propenoic acids, was synthesized as a part of our research in the antitubercular field, according to an international program with the Tuberculosis Antimicrobial Acquisition and Coordinating Facility (TAACF). This work reports the preparation and analytical and spectroscopic characterization (MS, UV, IR, 1H NMR) of all compounds synthesized. Among these only a few compounds (E-4b,c, E-5a, E-7e and E-8d) were found to be endowed with modest growth inhibition of Mycobacterium tuberculosis. However, the obtained results allowed to acquire interesting structure-activity relationships. PMID- 11902650 TI - Titrimetric and spectrophotometric assay of some antihistamines through the determination of the chloride of their hydrochlorides. AB - Two simple, rapid and reliable methods for the determination of four antihistamines based on the measurement of the chloride of their hydrochlorides are described. In the titrimetric method, the chloride content of each drug is determined by titrating with mercury(II) nitrate using diphenylcarbazone bromothymol blue as indicator. In the spectrophotometric method, to a fixed concentration of mercury(II)-diphenylcarbazone complex different amounts of drug are added and the decrease in absorbance of mercury(II)-diphenylcarbazone complex, consequent to the replacement of diphenylcarbazone of the complex by the chloride of the drug, was measured at 540 nm. The stoichiometry of the reaction that forms the basis for titrimetry is assessed. Different variables affecting the color formation in spectrophotometry were studied and optimized. At the wavelength of maximum absorption, Beer's law is obeyed in the 0-100 microg ml(-1) range. The molar absorptivity and Sandell sensitivity are calculated. The proposed methods were applied for the analysis of pharmaceutical formulations containing these drugs. Statistical treatment of the experimental results indicates that the procedures are precise and accurate. Excipients used as additives in pharmaceutical formulations did not interfere in the proposed procedures as shown by the recovery studies. PMID- 11902652 TI - Synthesis of variously 9,9-dialkylated octahydropyrimido [3,4-a]-s-triazines with potential antifungal activity. AB - 9,9-Dialkyloctahydropyrimido[3,4-a]-s-triazines were synthesized by iminodimethylation reaction between a 5,5-dialkyl-6-aminopyrimidine-2.4(3H,5H) dione, a substituted aniline and two moles of formaldehyde. The synthesis of.5,5 dialkyl-6-aminopy-rimidinedione consisted of the condensation of urea with ethyl 2,2-dialkylcyanoacetates. 18 Octahydropyrimido[3,4-a]-s-triazines were synthesized and compounds resulting from a supplementary aminomethylation were also obtained. Most of these compounds were tested for antifungal activity in vitro. Only 9.9-dibutyl-6,8-dioxo-3(2-chlorophenyl)2,3,4,5,6,7,8,9 octahydropyrimido[3,4-a]-s-triazine showed some activity against Microsporum canis. PMID- 11902651 TI - Synthesis and anti-inflammatory activity of 1-acylthiosemicarbazides, 1,3,4 oxadiazoles, 1,3,4-thiadiazoles and 1,2,4-triazole-3-thiones. AB - Sixteen 1-(2-naphthyloxyacetyl)-4-substituted-3-thiosemicarbazide, 2-(2 naphthyloxymethyl)-5-substitutedamino-1,3,4-oxadiazole, 2-(2-naphthyloxymethyl)-5 substitutedamino-1,3,4-thiadiazole and 5-(2-naphthyloxymethyl)-4-substituted 1,2,4-triazole-3thione derivatives have been prepared and evaluated as orally active anti-inflammatory agents with reduced side-effects. The structures of the compounds were confirmed by IR and 1H NMR spectral data and microanalysis. The anti-inflammatory and ulcerogenic activities of the compounds were compared with naproxen, indomethacin and phenylbutazone. In carrageenan-induced foot pad edema assay, 2-(2-naphthyloxymethyl)-5-methylamino-1,3,4-oxadiazole, 5-(2 naphthyloxymethyl)-4-methyl-1,2,4-triazole-3-thione and 5-(2-naphthyloxymethyl)-4 ethyl-1,2,4-triazole-3-thione showed an interesting anti-inflammatory activity. In the air-pouch test, 1,3,4-oxadiazole and 1,2,4-triazole-3-thione derivatives reduced total number of leukocytes of the exudate that indicates excellent inhibition of prostaglandin production. Side effects of the compounds were examined on gastric mucosa, liver and stomach and none of the compounds showed significant side effects compared with reference nonsteroidal anti-inflammatory drugs (NSAIDs). PMID- 11902653 TI - A study of the conditions of the supercritical fluid extraction in the analysis of selected anti-inflammatory drugs in plasma. AB - Supercritical fluid extraction (SFE) was employed to analyze selected anti inflammatory drugs in plasma. Evaluation of selected drugs (ibuprofen, indomethacin, and flufenamic acid) was performed using the HPLC method on columns with the reverse phase C-18 and detection in the UV region of the spectrum. A study of the conditions of SFE carried out for 30 min at 50 degrees C investigated the magnitude of the pressure of carbon dioxide suitable for drug extraction, the selection of the collecting solvent, and the modification of CO2 with an organic solvent. The results of the study made it possible to determine the optimal procedure for SFE of ibuprofen, indomethacin, and flufenamic acid from plasma, which renders their HPLC quantification possible. PMID- 11902654 TI - Synthesis and calcium channel antagonist activities of 3-nitrooxyalkyl, 5-alkyl 1,4-dihydro-2,6-dimethyl-4-(1-methyl-5-nitro-2-imidazolyl)-3, 5 pyridinedicarboxylates. AB - A group of racemic 3-[(2-nitrooxyethyl), (3-nitrooxypropyl), (4-nitrooxybutyl) or (1,3-dinitrooxy-2-propyl)], 5-methyl (ethyl or propyl) 1,4-dihydro-2,6-dimethyl-4 (1-methyl-5-nitro-2-imidazolyl)-3,5-pyridinedicarboxylates (18-29) were synthesized using modified Hantzsch reaction that involved the condensation of 2 nitrooxyethyl (8), 3-nitrooxypropyl (9), 4-nitrooxybutyl (10) or 1,3-dinitrooxy-2 propyl (13) acetoacetate with methyl (14), ethyl (15) or isopropyl (16) 3 aminocrotonate and 1-methyl-5-nitroimidazole-2-carboxaldehyde (17). In vitro calcium channel antagonist activities were determined using a guinea pig ileum longitudinal smooth muscle assay. Compounds 18-29 exhibited superior, or equipotent, calcium antagonist activity (IC50= 10(11) - 10(-13) M range) relative to the reference drug nifedipine (IC50 = 1.07 +/- 0.12 x 10(-11) M), which could serve as potential probes to investigate the in vivo release of nitric oxide which induces vascular muscle relaxation. PMID- 11902655 TI - Design and development of 2,3-benzodiazepine (CFM) noncompetitive AMPA receptor antagonists. AB - 2,3-Benzodiazepines represent a class of heterocyclic compounds that interact with AMPA-type glutamate receptors in a noncompetitive manner. These compounds have attracted great interest for their pharmacological effects against acute and chronic neurodegenerative diseases, such as ischemia and epilepsy. We synthesized a large number of 2,3-benzodiazepine derivatives, which showed anticonvulsant properties in different seizure models and a noncompetitive blockade of AMPA receptor. This article will briefly mention our work in this field and the main SAR considerations. PMID- 11902656 TI - Ring substituted 3-phenyl-1-(2-pyrazinyl)-2-propen-1-ones as potential photosynthesis-inhibiting, antifungal and antimycobacterial agents. AB - Four series of ring substituted (E)-3-phenyl-1-(2-pyrazinyl)-2-propen-1-ones were prepared by means of modified Claisen-Schmidt condensation of acetylpyrazines with aromatic aldehydes. The structures were confirmed by elemental analysis, IR, 1H NMR and 13C NMR spectra. The compounds were tested for specific biological properties and some derivatives exhibited photosynthesis-inhibiting, antifungal and antimycobacterial properties. The most pronounced effects were observed with compounds substituted with phenolic groups. Ortho-hydroxyl substituted derivatives were more potent than the corresponding para-hydroxyl substituted analogues. PMID- 11902657 TI - Synthesis of some 1,2,4-triazolo[3,2-b]-1,3-thiazine-7-ones with potential analgesic and antiinflammatory activities. AB - Starting from 3-substituted-1,2,4-triazole-5-thiones (la-h), eight new 5 carbomethoxy-2-substituted-7H-1,2,4-triazolo[3,2-b]-1,3-thiazine-7-ones (2a-h) were synthesized and characterized by spectral and elementary analysis. The obtained compounds were submitted to preliminary pharmacological assay to evaluate their antiinflammatory and analgesic activities as well as gastrointestinal irritation liability and acute toxicity. Among the compounds studied, compounds 2c, 2d, 2e and 2h showed most remarkable antiinflammatory activity in the carrageenan and serotonin induced edema and in the inhibition of castor oil-induced diarrhea tests. The analgesic activity of these active compounds correlated with their antiinflammatory activities in the inhibition of acetic acid-induced writhing test. In gastric ulceration studies, the compounds were found safety at low dose levels (10 and 20 mg/kg). PMID- 11902658 TI - Molecular modeling as a powerful technique for understanding small-large molecules interactions. AB - In the present review we summarize recent work, aimed at a better understanding of the interactions in macromolecule ligand complexes, performed by means of computational tools such as pseudoreceptor generation, molecular docking, conformational search and energy minimization. While the first approach has been applied when the three-dimensional structural properties of the biological target were unknown, the remaining protocols exploited the knowledge of the overall structure of the involved macromolecules and their active sites. Molecular modeling techniques were used in the cases reported to study and propose macromolecular binding sites and to predict their interactions with bioactive conformers of the ligands. PMID- 11902660 TI - Synthesis and antimicrobial activity of some new hydrazones of 4-fluorobenzoic acid hydrazide and 3-acetyl-2,5-disubstituted-1,3,4-oxadiazolines. AB - A series of hydrazide hydrazones and 1,3,4-oxadiazolines of 4-fluorobenzoic acid hydrazide were prepared and evaluated as potential antimicrobial agents and were tested for their antibacterial and antifungal activities against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. From these compounds, 4-fluorobenzoic acid[(5-nitro-2-furanyl)methylene]hydrazide (1a) showed equal activity with ceftriaxone against S. aureus. In addition, the MIC values of compounds 1c, 1d and 2a for the same strain were in the range of those reported for ceftriaxone according to NCCLS 1997. PMID- 11902659 TI - 30-day intravenous administration of VRCTC-310-ONCO in rabbits. AB - VRCTC-310-ONCO, an agent based on the snake phospholipase A2 (crotoxin), is currently under clinical development. After phase I study in patients by intramuscular administration, the interest of intravenous (IV) dosing arose. To evaluate IV administration of VRCTC-310-ONCO in rabbits, ten animals were subjected to surgical implant of fixed jugular catheter, by which they received daily IV doses of 0.03 mg/kg body weight of VRCTC-310-ONCO for 30 days (n = 8) or saline (n = 2). The procedure was well tolerated in all rabbits. One of the animals died after the sixth dose of VRCTC-310-ONCO with CNS involvement; two additional rabbits required dose-reduction. All other rabbits achieved 30 days of treatment and were sacrificed. All rabbits (even controls) developed lymphocytosis and mild anaemia, without changes in blood neutrophils. No changes were found in serum transaminases (GOT and GPT), cholesterol, triglycerides, and y-glutamyl transpeptidase. At necropsy, chronic granulation tissue was found surrounding the implant in all rabbits. VRCTC-3 10-ONCO-treated rabbits presented generalised and marked swelling of hepatocytes, with areas of cytoplasmic vacuolisation. No abnormalities were found in kidney, heart, lung, spleen, adrenal gland, uterus, testes and ovary. Additional studies with IV route for VRCTC-310-ONCO, including humans, are required to define its toxicity in the clinical setting. PMID- 11902661 TI - Isoxazolo[3,4-d]pyridazinones and analogues as Leishmania mexicana PDE inhibitors. AB - A series of isoxazolopyridazinones and analogues has been prepared and evaluated as Leishmania mexicana phosphodiesterase (PDE) inhibitors. Some of the synthesized compounds showed a moderate PDE inhibitory activity at 100 microM and preliminary structure-activity relationships were discussed. PMID- 11902662 TI - 2-Diazoindoles: building blocks for the synthesis of antineoplastic agents. AB - 2-Diazoindoles were prepared by diazotization of the corresponding 2-aminoindoles followed by neutralisation. 2-Diazoindoles were utilised for the synthesis of 2 triazenoindoles, indolo[2,1-d][1,2,3,5]tetrazines and indolo[2,1 c][1,2,4]triazines. Most of these compounds exhibited in vitro antiproliferative activity. PMID- 11902664 TI - Oxidative folding of bovine pancreatic ribonuclease A: insight into the overall catalysis of the refolding pathway by phosphate. AB - The effects of the strong stabilizing anion, phosphate, on the oxidative folding of bovine pancreatic ribonuclease A were examined. Phosphate was found to catalyze several steps involved in the oxidative folding process at pH 8.0 and 25 degrees C, resulting in an increase in the rate of pre-equilibration of unstructured species on the folding pathway. In the presence of 400 mM phosphate, the overall increase in the rate of regeneration of native protein was caused primarily by the increased formation and stabilization of tertiary structure in the nativelike intermediates, des-[40-95] and des-[65-72], involved in the rate determining step. Based on the regeneration of native protein and the stability of Cys--> Ala substituted mutant analogs of the des-species, (C40A, C95A) and (C65A, C72A), it is suggested that the primary role of phosphate is to catalyze the overall regeneration of native protein through nonspecific electrostatic and hydrogen-bonding effects on the protein and solvent. PMID- 11902663 TI - Identification of functionally important cysteines in the alpha-subunit of transducin by chemical cross-linking techniques. AB - Transducin (T), the G-protein in the visual system, is a heterotrimer arranged as two functional units, Talpha and Tbetagamma. N,N'-1,2-phenylenedimaleimide (o PDM) and N,N'-1,4-phenylenedimaleimide (p-PDM), two cysteine specific homobifunctional agents, were used to covalently cross-link T and its units. A complete inhibition in T function was observed in the presence of these compounds. Incubation of Talpha with o-PDM or p-PDM resulted in the formation of high-molecular-weight oligomers of 70-, 105-, 140-, and >200 kDa, as well as intramolecular cross-linked polypeptides that migrated as 35- and 37-kDa bands. Additionally, the treatment of Tbetagamma with both reagents produced a major species of 46-kDa. The combination of intact Talpha and o-PDM- or p-PDM-treated Tbetagamma reconstituted T native activities. On the contrary, when o-PDM- or p PDM-modified Talpha was incubated with intact Tbetagamma, more than 90% inhibition on T function was observed. Hence, the cysteines modified and/or cross linked on Talpha represent functionally important residues of T. PMID- 11902665 TI - Glycerol-induced formation of the molten globule from acid-denatured cytochrome c: implication for hierarchical folding. AB - At high concentration (98% or higher, v/v), glycerol induces collapse of acid denatured cytochrome c into a compact state, the G(U) state, showing a molten globule character. The G(U) state possesses a nativelike alpha-helix structure but a tertiary conformation less packed with respect to the native state. The spectroscopic properties of the G(U) state closely resemble those of the molten globule stabilized by the organic solvent from the native protein (called the G(N) state), indicating that glycerol can stabilize the molten globule of cytochrome c either from the native or the acid-denatured protein. The G(U) and the G(N) states show spectroscopic (and, thus, structural) properties and stabilities comparable to those of molten globules stabilized by different effectors, despite the fact that the mechanisms involved in the molten globule formation may significantly differ. This implies in cytochrome c a hierarchy for the rupture (native-to-molten globule) or the formation (unfolded-to-molten globule) of intramolecular interactions leading to the stabilization of the molten globule state of the protein, independently from the effector responsible for the structural transition, in accord with the sequential model proposed by Englander and collaborators. PMID- 11902666 TI - Primary structure characterization of Bothrops jararacussu snake venom lectin. AB - The complete amino acid sequence of the lectin from Bothrops jararacussu snake venom (BJcuL) is reported. The sequence was determined by Edman degradation and amino acid analysis of the S-carboxymethylated BJcuL derivative (RC-BJcuL) and from its peptides originated from enzymatic digestion. The sequence of amino acid residues showed that this lectin displays the invariant amino acid residues characterized in C-type lectins. Amino acids analysis revealed a high content of acidic amino acids and leucine. These findings suggest that BJcuL, like other snake venom lectins, possesses structural similarities to the carbohydrate recognition domain (CRD) of calcium-dependent animal lectins belonging to the C type beta-galactoside binding lectin family. PMID- 11902667 TI - Diethylpyrocarbonate inhibition of vacuolar H+-pyrophosphatase possibly involves a histidine residue. AB - Vacuolar proton pumping pyrophosphatase (H+-PPase; EC 3.6.1.1) plays a pivotal role in electrogenic translocation of protons from cytosol to the vacuolar lumen at the expense of PPi hydrolysis. A histidine-specific modifier, diethylpyrocarbonate (DEPC), could substantially inhibit enzymic activity and H+ translocation of vacuolar H+-PPase in a concentration-dependent manner. Absorbance of vacuolar H+-PPase at 240 nm was increased upon incubation with DEPC, demonstrating that an N-carbethoxyhistidine moiety was probably formed. On the other hand, hydroxylamine, a reagent that can deacylate N carbethoxyhistidine, could reverse the absorption change at 240 nm and partially restore PPi hydrolysis activity as well. The pKa of modified residues of the enzyme was determined to be 6.4, a value close to that of histidine. Thus, we speculate that inhibition of vacuolar H+-PPase by DEPC possibly could be attributed to the modification of histidyl residues on the enzyme. Furthermore, inhibition of vacuolar H+-PPase by DEPC follows pseudo-first-order rate kinetics. A reaction order of 0.85 was calculated from a double logarithmic plot of the apparent reaction constant against DEPC concentration, suggesting that the modification of one single histidine residue on the enzyme suffices to inhibit vacuolar H+-PPase. Inhibition of vacuolar H+-PPase by DEPC changes Vmax but not Km values. Moreover, DEPC inhibition of vacuolar H+-PPase could be substantially protected against by its physiological substrate, Mg2+-PPi. These results indicated that DEPC specifically competes with the substrate at the active site and the DEPC-labeled histidine residue might locate in or near the catalytic domain of the enzyme. Besides, pretreatment of the enzyme with N-ethylmaleimide decreased the degree of subsequent labeling of H+-PPase by DEPC. Taken together, we suggest that vacuolar H+-PPase likely contains a substrate-protectable histidine residue contributing to the inhibition of its activity by DEPC, and this histidine residue may located in a domain sensitive to the modification of Cys-629 by NEM. PMID- 11902668 TI - Cofactors in sarcosine oxidase from Corynebacterium sp. U-96. AB - Sarcosine oxidase from Corynebacterium sp. U-96 is a heterotetrameric enzyme that was reported to contain 1 mol of covalently bound FAD and 1 mol of non-covalently bound FAD. This work describes the result of reinvestigation of the cofactors in this enzyme. The enzyme was found to contain 1 mol of non-covalently-bound NAD+, 1 mol of non-covalently-bound FAD, and 1 mol of covalent FMN. The covalent FMN was identified by the mass and amino acid sequence analyses of the flavin peptide. PMID- 11902669 TI - Evaluation of hydrophobicity versus chaperonelike activity of bovine alphaA- and alphaB-crystallin. AB - Calf lens alphaA-crystallin isolated by reversed-phase HPLC demonstrates a slightly more hydrophobic profile than alphaB-crystallin. Fluorescent probes in addition to bis-ANS, like cis-parinaric acid (PA) and pyrene, show higher quantum yields or Ham ratios when bound to alphaA-crystallin than to alphaB-crystallin at room temperature. Bis-ANS binding to both alphaA- and alphaB-crystallin decreases with increase in temperature. At room temperature, the chaperone-like activity of alphaA-crystallin is lower than that of alphaB-crystallin whereas at higher temperatures, alphaA-crystallin shows significantly higher protection against aggregation of substrate proteins compared to alphaB-crystallin. Therefore, calf lens alphaA-crystallin is more hydrophobic than alphaB-crystallin and chaperone like activity of alpha-crystallin subunits is not quantitatively related to their hydrophobicity. PMID- 11902670 TI - Primary structures of alpha- and beta-subunits of alpha-amylase inhibitors from seeds of three cultivars of Phaseolus beans. AB - The primary structures of three alpha-amylase inhibitors (TAI, DAI, and MAI-2) consisting of glycoprotein subunits alpha and beta from the respective seeds of three cultivars of Phaseolus beans, Toramame (Phaseolus vulgaris L.), Daifukumame (Phaseolus vulgaris L.), and Murasakihanamame (Phaseolus coccineus L.) were determined by sequencing the peptide fragments derived from their enzymatic digestions. Major sugar chains of the inhibitors were also assessed by analyzing glycopeptides in the enzymatic digests. The subunits, alpha and beta, were shown to be composed of 76 and 139 amino acid residues, respectively, in each inhibitor. The overall amino acid sequences of the inhibitors were slightly different from one another. Furthermore, the sequence of TAI was the same as that deduced from a cDNA clone encording alpha-amylase inhibitor-1 from the common bean (Phaseolus vulgaris L.). It was also revealed that there were two N glycosylation sites in each alpha-subunit: PA-derivatives of the major N-glycans were estimated to be M6B at Asn(12) and M9A at Asn(65). Each beta-subunit of TAI and MAI-2 had two N-glycosylation sites, while the beta-subunit of DAI had only one site. The major N-glycans pyridylaminated were estimated to be M3X at Asn(63) in each beta-subunit and M3FX at Asn(83) in beta-subunits of TAI and MAI-2. PMID- 11902671 TI - PPAR expression and function during vertebrate development. AB - The peroxisome proliferator activated receptors (PPARs) are ligand activated receptors which belong to the nuclear hormone receptor family. As with other members of this superfamily, it is thought that the ability of PPAR to bind to a ligand was acquired during metazoan evolution. Three different PPAR isotypes (PPARalpha, PPARbeta, also called 6, and PPARgamma) have been identified in various species. Upon binding to an activator, these receptors stimulate the expression of target genes implicated in important metabolic pathways. The present article is a review of PPAR expression and involvement in some aspects of Xenopus laevis and rodent embryonic development. PPARalpha and beta are ubiquitously expressed in Xenopus early embryos but become more tissue restricted later in development. In rodents, PPARalpha, PPARbeta and PPARgamma show specific time- and tissue-dependent patterns of expression during fetal development and in the adult animals. PPARs are implicated in several aspects of tissue differentiation and rodent development, such as differentiation of the adipose tissue, brain, placenta and skin. Particular attention is given to studies undertaken by us and others on the implication of PPARalpha and beta in rodent epidermal differentiation. PMID- 11902673 TI - Regulation of cell migration during tracheal development in Drosophila melanogaster. AB - Most of the knowledge concerning the intracellular mechanisms involved in cell locomotion have been obtained from in vitro studies of cells in culture. Many of the concepts derived from these studies have been partially confirmed in in vivo systems but numerous questions regarding the developmental control of cell migration remain to be addressed. Tracheal morphogenesis in Drosophila melanogaster embryos represents an in vivo model system to study the genetic control of cell migration. We review what is known about tracheal development and regulation of tracheal cell migration. We try to link these in vivo studies and the movement of cells over two dimensional substrates and elaborate on important questions which remain to be addressed in the future. PMID- 11902672 TI - Loss and gain of domains during evolution of cut superclass homeobox genes. AB - The cut superclass of homeobox genes has been divided into three classes: CUX, ONECUT and SATB. Given the various completed genomes, we have now made a comprehensive survey. We find that there are only two cut domain containing genes in Drosophila, one CUX and one ONECUT type. Caenorhabditis elegans has undergone an expansion of the ONECUT subclass genes and has a gene cluster with three ONECUT class genes, one of which has lost the cut domain. Two of these genes contain a conserved sequence motif, termed OCAM, which also occurs in another gene in C. elegans this motif seems to be nematode specific. A recently uncovered C. elegans CUX gene has sequence conservation in its amino-terminus with vertebrate CUX proteins. Further, the 5' end of this gene containing the conserved region can undergo alternative splicing to give rise to a protein with a different carboxy-terminus lacking the cut- and homeodomain. This protein is conserved in its entirety with vertebrate genes termed CASP--which are also alternative splice products of the CUX genes--and with plant and fungal genes. The highly divergent SATB genes share a conserved amino terminal domain, COMPASS, with the Drosophila defective proventriculus gene and a C. elegans ORF. These two "COMPASS" family genes encode two highly divergent homeodomains, may be homologues of the SATB genes and thus probably belong to the cut superclass, too. PMID- 11902674 TI - Extensive conservation of sequences and chromatin structures in the bxd polycomb response element among Drosophilid species. AB - The Polycomb Response Element (PRE) is the nucleation site for the Polycomb silencing complexes. The sequences responsible for the recruitment of the components of the Polycomb complex are not well understood. A comparison of the bxd PRE sequences from several different Drosophila species shows that some changes have occurred during phylogeny but large blocks of sequence are conserved after a divergence of some 60 million years. We compare the PRE sequences, the sites of some known PRE binding proteins, the conservation of DNasel hypersensitive sites and relate them to the sequence of the Ultrabithorax promoter which these PREs regulate. PMID- 11902675 TI - New telomere formation during the process of chromatin diminution in Ascaris suum. AB - Chromatin diminution in the parasitic nematode Ascaris suum represents an interesting case of developmentally programmed DNA rearrangement in higher eukaryotes. At the molecular level, it is a rather complex event including chromosome breakage, new telomere formation and DNA degradation. Analysis of a cloned somatic telomere (pTel1) revealed that it has been newly created during the process of chromatin diminution by the addition of telomeric repeats (TTAGGC)n to a chromosomal breakage site (Muller et al., 1991). However, telomere addition does not occur at a single chromosomal locus, but at many different sites within a short chromosomal region, termed CBR1 (chromosomal breakage region 1). Here we present the cloning and the analysis of 83 different PCR amplified telomere addition sites from the region of CBR1. The lack of any obvious sequence homology shared among them argues for a telomerase-mediated healing process, rather than for a recombinational event. This hypothesis is strongly supported by the existence of 1-6 nucleotides corresponding to and being in frame with the newly added telomeric repeats at almost all of the telomere addition sites. Furthermore, we show that telomeres are not only added to the ends of the retained chromosomal portions, but also to the eliminated part of the chromosomes, which later on become degraded in the cytoplasm. This result suggests that de novo telomere formation during the process of chromatin diminution represents a non-specific process which can heal any broken DNA end. PMID- 11902676 TI - Maternal-effect loci involved in Drosophila oogenesis and embryogenesis: P element-induced mutations on the third chromosome. AB - A collection of 1609 recessive P-lethal mutations on the third chromosome was tested in germline clones for effects on egg differentiation and embryonic development. In 164 lines, normal differentiation of the egg chamber is prevented and in 841 lines, embryos develop abnormally. This latter group of maternal effect mutations was subdivided into 23 classes based on the cuticular phenotypes. Our collection comprises new alleles of previously characterized genes (e.g. kayak, punt, string, tramtrack). For some of the genes identified in this screen, a maternal contribution to embryonic development has not been described previously (e.g. extramacrochaete, Trithorax-like, single minded, couch potato, canoe). The genes classified in our study with a dual function during oogenesis and embryogenesis not only substantially extends the existing collection of maternal-effect genes but will also aid further understanding of how patterning of the Drosophila embryo is controlled by the maternal genome. PMID- 11902677 TI - The beginnings of developmental biology in Swiss universities. AB - This contribution describes the pioneer work in Developmental Biology, initiated by Swiss scientists. The anatomist W. His (1831-1904) deserves credit as the founder of "Descriptive Embryology". Using novel microscopical techniques, he documented the formation of the embryonic body of various vertebrates as an approach to reveal the mechanisms of morphogenesis. Based on studies of the chick embryo, he designed a fate map of the germ-disk, comprising organ-forming regions, from which the corresponding embryonic structures originate. F. Baltzer (1886-1974) initiated original studies on the role of nuclear and cytoplasmic factors in the development of sea urchin hybrids and newt merogons. Through his experiments on interspecific and intergeneric chimeras of amphibians, he further contributed to the emerging field of "Developmental Genetics". F.E. Lehmann (1901 70) inaugurated work on "Chemical Embryology" and later moved to "Cell Biology". His discoveries on stage- and regional-specific inhibition of morphogenesis by LiCl in newt embryos were important for the understanding of malformations. Further studies regarding the action of cytostatic substances on tail regeneration in tadpoles were intended to yield information on growth control. Original contributions to "Experimental Embryology" were also made by R. Geigy (1902-1995), who devised an ingenuous procedure for obtaining sterile Drosophila flies by UV-irradiation of eggs. In later studies on anuran metamorphosis, he discovered that the morphogenetic effects of the metamorphic hormones are organ specific and that competence of the larval tissues to respond to thyroid hormone is stage-dependent. PMID- 11902678 TI - mgm 1, the earliest sex-specific germline marker in Drosophila, reflects expression of the gene esg in male stem cells. AB - The pathway that controls sex in Drosophila has been well characterized. The elements of this genetic hierarchy act cell-autonomously in somatic cells. We have previously shown that the sex of germ cells is determined by a different mechanism and that somatic and autonomously acting elements interact to control the choice between spermatogenesis and oogenesis. A target for both types of signals is the enhancer-trap mgm1, which monitors male-specific gene expression in germ cells. Here we report that mgm1 reflects the expression of escargot (esg), a member of the snail gene family, which are transcription factors with zink finger motifs. Genes of this family partially redundantly control a number of processes involving cell fate choices. The regulation of gene expression in germ cells by sex-specific esg enhancers is already seen in embryos. Therefore, autonomous and non-autonomous sex-specific factors that participate in germline sex determination are already present at this early stage. esg is expressed in the male gonad, both in somatic cells and in germline stem cells. We show that esg expression in the male germline is not required for proper sex determination and spermatogenesis, as functional sperm is differentiated by mutant germ cells in wild type hosts. However, somatic esg expression is required for the maintenance of male germline stem cells. PMID- 11902679 TI - A new gene in Drosophila melanogaster, Ravus, the phantom of the modifier of position-effect variegation Su(var)3-7. AB - In a search for homologues of the dominant modifier of position-effect variegation Su(var)3-7, we have identified one ORF in Drosophila melanogaster. The 359 amino acid deduced protein is much shorter than the 1169 amino acid protein Su(var)3-7. Surprisingly, the two genes are very close to each other at 87E on the polytene chromosome map, and are transcribed divergently. The triplet coding for the N-terminus amino acid of the new gene lies only 368 base pairs from the start of transcription of Su(var)3-7. This opposite orientation of the homologue has led us to name it Ravus. The N-terminus of the Ravus protein contains only one of the seven unusual zinc fingers of Su(var)3-7. A second region of similarity encodes an acidic domain. Finally, there is a block of high similarity near the C-terminus of the two proteins. It corresponds to a new conserved protein domain, BESS, found also in the BEAF and Stonewall Drosophila proteins. We have constructed a tagged Ravus protein, and have expressed it as a heat-shock inducible transgene. Ravus associates in vivo with polytene chromosomes but, in contrast to the heterochromatin-associated protein Su(var)3 7, does not show specificity for the chromocenter. Ravus does not seem either to modify the genomic silencing of position-effect variegation, as over-expression of the transgene does not affect the variegated phenotype of a number of rearrangements tested. PMID- 11902680 TI - A screen for genes expressed in Drosophila imaginal discs. AB - The development of Drosophila imaginal discs serves as a model system to understand how genes determine the shape and size of an organ. The identification of genes involved in this process is an important step towards this goal. Here we describe a P-element based enhancer trap screen for genes expressed in the larval imaginal discs. Our aim was to establish a large collection of enhancer trap lines each showing expression of Gal4 in imaginal discs. To this end, we improved the well established P-element vector pGawB in order to obtain higher in vivo transposition frequencies. In addition we chose an F1-screening approach using UAS-GFP as a reporter gene. This system permits the efficient screening of larval and pupal stages of living animals and the detection of imaginal gene expression patterns through the transparent cuticle. The procedure has been optimized for high-throughput. 2'000 P-element insertions have been established which exhibit expression in imaginal discs. PMID- 11902682 TI - A nested deletion approach to generate Cre deleter mice with progressive Hox profiles. AB - In mice, the loxP/Cre recombinase-dependent system of recombination offers powerful possibilities for engineering genetic configurations of interest. This system can also be advantageously used for conditional mutagenesis in vivo, whenever such an approach is required due to deleterious effects of either one mutation, or a combination thereof. Here, we report on the production of an allelic series of insertions of a Hoxd11/Cre fusion transgene at different positions within the HoxD complex, in order to produce the CRE recombinase with a 'Hox profile' progressively more extended. We used the R26R (R26R) reporter mouse line to functionally assess the distribution and efficiency of the CRE enzyme and discuss the usefulness of these various lines as deleter strains. PMID- 11902681 TI - Embryonic expression of Xenopus SGLT-1L, a novel member of the solute carrier family 5 (SLC5), is confined to tubules of the pronephric kidney. AB - Plasma membrane proteins of the solute carrier family 5 (SLC5) are responsible for sodium-coupled uptake of ions, sugars and nutrients in the vertebrate body. Mutations in SLC5 genes are the cause of several inherited human disorders. We have recently reported the cloning and transport properties of SGLT-1L, a Xenopus homologue of the sodium-dependent glucose cotransporter 1 (SGLT-1) [Nagata et al. (1999) Am. J. Physiol. 276: G1251 -G 1259]. Here, we describe the phylogenetic relationship of SGLT-1L with other members of the SLC5 family and characterize its expression during Xenopus embryogenesis and in organ cultures. Sequence comparisons and phylogenetic analyses of all known vertebrate SLC5 sequences indicated that Xenopus SGLT-1L encodes a novel SLC5 member, which shares highest amino acid identity with mammalian ST-1 proteins. Temporal and spatial expression of SGLT-1L during Xenopus embryogenesis was examined by whole mount in situ hybridization. Initiation of SGLT-1L expression occurred in the late tailbud embryo. Remarkably, expression was restricted to the developing pronephric kidney. SGLT-1L was highly expressed in tubular epithelia, but completely absent from the epithelia of the duct. Analysis of growth factor-treated animal caps indicated that expression of SGLT-1L could also be induced in organ cultures. Taken together, our findings indicate that the expression of sodium-dependent solute cotransporter genes in early segments of the excretory system appears to be conserved between pronephric and metanephric kidneys. Furthermore, we establish SGLT-1L as a novel, highly specific molecular marker for pronephric tubule epithelia undergoing maturation and terminal differentiation in Xenopus. PMID- 11902684 TI - Ernst Hadorn, a pioneer of developmental genetics. AB - This article gives a short and personal portrait of Ernst Hadorn (1902-1976), one of the most influential developmental biologists in Europe. Hadorn initially worked with amphibia, but then soon turned to Drosophila where he very successfully studied lethal factors and the development of imaginal disks. PMID- 11902683 TI - Cell-intrinsic and cell-extrinsic cues regulating lineage decisions in multipotent neural crest-derived progenitor cells. AB - Multipotent stem cells must generate various differentiated cell types in correct number and sequence during neural development. In the peripheral nervous system (PNS), this involves the formation of postmigratory progenitor cell types which maintain multipotency and are able to give rise to neural and non-neural cells in response to instructive growth factors. We propose that fate restrictions in such progenitor cells are controlled by the combinatorial interaction of different extracellular signals, including community effects in response to both neurogenic and gliogenic factors. In addition, distinct progenitor cell types display intrinsic differences which modulate their response to the extracellular environment. Thus, a progenitor cell is apparently able to integrate multiple intrinsic and extrinsic cues and thereby to choose fates appropriate for its location. Fate analysis of genetically modified progenitor cells will help to identify the molecules involved. This approach appears promising given the identification of multipotent progenitor cells from the mouse PNS and the availability of genetics in the mouse system. PMID- 11902685 TI - From transdetermination to the homeodomain at atomic resolution. An interview with Walter J. Gehring. PMID- 11902686 TI - Cnidarians as a model system for understanding evolution and regeneration. AB - Hydra and Podocolyne are two cnidarian animals which provide complementary advantages for analysing developmental mechanisms possibly reflecting the basic developmental processes shared by most bilaterians. Interestingly, these mechanisms remain accessible all along the life of these animals, which bud and regenerate, whatever their age. The Hydra polyp permits a direct study of the molecular cascades linking amputation to regeneration. Podocoryne displays a complete life cycle, polyp and medusa stages with a fast and inducible sexual cycle and an unparalleled In vitro transdifferentiation potential. In both cases, a large number of evolutionarily conserved molecular markers are available, and analysis of their regulation highlights the molecular mechanisms which underly pattern formation in these two species. PMID- 11902687 TI - Xenopus helveticus, an endangered species? AB - This review traces the history of Xenopus research in Switzerland, its worldwide beginnings and British chapters having been summarised previously (Gurdon and Hopwood, 2000). As in other countries, Xenopus was initially used in the pharmaceutical industry at Basel for pregnancy testing. Developmental biologists became interested in this peculiar amphibian because it may be induced to ovulate all year round. Swiss Xenopus research is reviewed over 50 years, from the introduction of Xenopus by Rudolf Weber to the University of Bern, the return from Great Britain of two Swiss expatriates, Michail Fischberg and Max Birnstiel through the numerous pupils of the founder labs to the independently arisen Xenopus research units in the country. Besides developmental biology, Swiss Xenopus research engaged in immunology, genetics and cell biology, the latter focusing mainly on the oocyte. It set highlights in molecular biology by isolating some of the first eukaryotic genes and analysing their transcriptional regulation and post-transcriptional modifications through 'surrogate genetic' approaches in the oocyte system. An important line of research applied this system to study nuclear trafficking. Presently, functional testing mainly serves to characterise the function of proteins produced from expression vectors injected into the oocyte. A main accent of developmental studies was, from the early beginnings and still today, set on molecular characterisation of gene function in the embryo. PMID- 11902688 TI - Developmental biology in Geneva: a three century-long tradition. AB - It was in the first half of the 18th century when life sciences started to flourish in the independent republic of Geneva. However, it is difficult to identify a genuine school of developmental biologists during that era. Nevertheless, several prominent scientists over the past two and a half centuries have established and maintained a strong tradition of studies in embryological development and reproduction. In this short historical account, we briefly pay tribute to these famous forerunners, by emphasizing both the originality and quality of their work, as well as the many accompanying conceptual and methodological advances. We start with Abraham Trembley (1710-1784) and the discovery of Hydra and of regeneration, and with Charles Bonnet (1720-1793) who, amongst other contributions, first observed parthenogenetic development. In the 19th century, Carl Vogt (1817-1895) and Edouard Claparede (1832-1871) were well known scientists in this field of research, whereas Hermann Fol (1845-1892) can be considered as one of the pioneers, if not the founder, of causal embryology, through his experiments on lateral asymmetry in manipulated chicken. More recently, Emile Guyenot (1885-1963) and Kitty Ponse (1897-1982) perpetuated this tradition, which is well alive nowadays in the city of Calvin. PMID- 11902689 TI - The genetic control of eye development and its implications for the evolution of the various eye-types. AB - Mutations in the Pax 6 homologs of mammals and insects prevent eye development and targeted expression of both mammal and insect Pax 6 homologs is capable of inducing functional ectopic eyes. Supported by RNA interference experiments in planarians and nemerteans, these findings indicate that Pax 6 is a universal master control gene for eye morphogenesis. Since all metazoan eyes use rhodopsin as a photoreceptor molecule and the same master control gene for eye development, we postulate a monophyletic origin of the various eye types. The finding of well developed eyes in jellyfish which essentially lack a brain, leads us to propose that the eye as a sensory organ evolved before the brain which is an information processing organ. The finding of highly developed eyes with a lens, vitreous body, stacked membranes like a retina and shielding pigment in unicellular dinoflagellates, raises the possibility that the prototypic eyes might have been acquired from symbionts. PMID- 11902690 TI - Musca domestica, a window on the evolution of sex-determining mechanisms in insects. AB - The genetic cascades regulating sex determination of the housefly, Musca domestica, and the fruitfly, Drosophila melanogaster, appear strikingly different. The bifunctional switch gene doublesex, however, is present at the bottom of the regulatory cascades of both species, and so is transformer-2, one of the genetic elements required for the sex-specific regulation of doublesex. The upstream regulators are different: Drosophila utilizes Sex-lethal to coordinate the control of sex determination and dosage compensation, i.e., the process that equilibrates the difference of two X chromosomes in females versus one X chromosome in males. In the housefly, Sex-lethal is not involved in sex determination, and dosage compensation, if existent at all, is not coupled with sexual differentiation. This allows for more adaptive plasticity in the housefly system. Accordingly, natural housefly populations can vary greatly in their mechanism of sex determination, and new types can be generated in the laboratory. PMID- 11902691 TI - Conserved genetic mechanisms for embryonic brain patterning. AB - A wealth of comparative embryological studies on the expression and function of homeotic genes and cephalic gap genes indicates that both gene groups are important for establishing and specifying the anteroposterior body axis during embryogenesis in bilaterian animals. Recently, studies of this kind have been extended to embryonic brain development in two genetic model systems, Drosophila and mouse. These studies demonstrate striking similarities in the pattern of expression and mode of action of these developmental control genes during embryonic patterning of the brain in both species. Thus, in both insect and mammalian species, members of the homeotic gene complex are involved in patterning the posterior brain anlage, where they control regionalized neuronal identity, and members of the cephalic gap genes, notably the otd/Otx gene family, are involved in patterning the anterior brain anlage where they control regionalized neurogenesis and neuronal identity. Furthermore, striking cross phylum rescue experiments show that insect and mammalian members of the orthodenticle gene family can functionally replace each other in embryonic brain and CNS patterning. Comparable cross-phylum rescue experiments have now also been carried out for the empty spiracles cephalic gap gene family. Taken together, these experiments suggest that the genetic mechanisms involved in embryonic brain development are conserved and indicative of a common evolutionary origin of the insect and vertebrate brain. For a more extensive and quantitative investigation of the molecular conservation of developmental mechanisms for brain patterning, functional genomic experiments are now underway in Drosophila. These experiments exploit the advent of sequenced genome information and the technology for large scale transcript imaging, with the goal of identifying the entire set of downstream genes which is under the control of these regulatory genes in embryonic brain development. PMID- 11902692 TI - Chromosome 21: a small land of fascinating disorders with unknown pathophysiology. AB - In the year 2000 we celebrated the sequencing of the entire long arm of human chromosome 21. This achievement now provides unprecedented opportunities to understand the molecular pathophysiology of trisomy 21, elucidate the mechanisms of all monogenic disorders of chromosome 21, and discover genes and functional sequence variations that predispose to common complex disorders. All of that requires the functional analysis of gene products in model organisms, and the determination of the sequence variation of this chromosome. PMID- 11902693 TI - Defining the cell lineages of the islets of Langerhans using transgenic mice. AB - In this Special Issue of the Int. J. Dev. Biol., we summarize our own studies on the development of the mouse endocrine pancreas, with special emphasis on the cell lineage relationships between the four islet cell types. Considerable knowledge concerning the ontogeny of the endocrine pancreas has been gained in recent years, mainly through the use of two complementary genetic approaches in mice: gene inactivation and genetic labelling of precursor cells. However, neither gene inactivation in KO mice nor co-localisation of hormones in single cells during development can be taken as evidence for cell lineage relationships among different cell types. The beta-cell lineage analysis was started by selectively ablating specific islet cell types in transgenic mice. We used the diphtheria toxin A subunit coding region under the control of insulin, glucagon or pancreatic polypeptide (PP) promoters, in order to eliminate insulin-, glucagon- or PP-expressing cells, respectively. Contrary to the common view, we demonstrated that glucagon cells are not precursors of insulin-producing cells. These results were in addition the first evidence of a close ontogenetic relationship between insulin and somatostatin cells. We pursued these analyses using a novel, more subtle approach: progenitor cell labelling through the expression of Cre recombinase in doubly transgenic mice. We were able to unequivocally establish that 1) adult glucagon- and insulin-producing cells derive from precursors which have never transcribed insulin or glucagon, respectively; 2) insulin cell progenitors, but not glucagon cell progenitors transcribe the PP gene and 3) adult glucagon cells derive from progenitors which do express pdx1. PMID- 11902695 TI - Adverse effects of smoking. Selected annotated bibliography. PMID- 11902694 TI - Cigarette smoking--a paradox. PMID- 11902696 TI - Bronchoscopy: coverage and reimbursement. PMID- 11902697 TI - Determinants of outcome for patients admitted to a long-term ventilator unit. AB - BACKGROUND: This prospective cohort study was done to identify determinants of successful weaning from mechanical ventilation among patients admitted to the 10 bed long-term ventilator unit (LTVU) of a teaching hospital. METHODS: Prospective patient surveillance and data collection were done on 472 patients admitted to the LTVU over a 4-year period (January 1996 to December 1999). RESULTS: Multiple logistic regression analysis showed that the absence of home mechanical ventilation at the time of hospital admission, absence of intensive care unit (ICU) readmission, and admission to the LTVU from a nonmedical service were independently associated with successful weaning. No statistical difference between hospital survivors and nonsurvivors was associated with length of stay in the LTVU and length of stay in the hospital. CONCLUSIONS: Patients admitted to an LTVU require prolonged hospitalizations and intensive resource utilization. These data suggest that improved methods for identifying patients who are unlikely to benefit from prolonged mechanical ventilation may assist physicians in their discussions with patients and family members as they consider various treatment options. PMID- 11902699 TI - Cardiac surgery in patients with moderate renal impairment. AB - BACKGROUND: There is a paucity of information concerning the results of cardiac surgery in patients with moderate impairment of renal function. We reviewed our recent experience to determine the results of operation and the long-term outcome. METHODS: Since January 1992, we have performed cardiac surgical procedures utilizing total cardiopulmonary bypass on 57 adult patients with preoperative serum creatinine values > or = 2.0 mg/dL and no history of dialysis. Operative procedures done were coronary artery bypass (39 patients), repeated coronary artery bypass (2), valve replacement with or without coronary artery bypass (12), and other procedures (4). RESULTS: No operative deaths occurred. There were 3 hospital deaths. Only 5 patients required perioperative dialysis; in 5 additional patients, chronic dialysis was begun from 4 to 24 months postoperatively. The surviving patients who were not receiving dialysis had a mean creatinine value of 2.4 mg/dL at most recent follow-up. CONCLUSIONS: Adult patients with moderate renal impairment can safely have major cardiac procedures. The majority of patients maintain stable renal function postoperatively. The overall results of cardiac surgery in this patient population are good. PMID- 11902698 TI - Accidental injection of epinephrine from an autoinjector: invasive treatment not always required. AB - BACKGROUND: Individual case reports of accidental injection with epinephrine appear in the literature and seem to represent the worst case scenarios. We present a case series of 28 exposures to epinephrine via autoinjector. METHOD: All accidental parenteral injections of epinephrine by autoinjector reported to two regional poison information centers over a 2-year period were included. RESULTS: Injection sites included digits (23 cases), palm (4 cases), and thigh (1 case). Symptoms included swelling, pallor, pain, and erythema. Four patients reported no effect, and 9 required no treatment. Ten patients obtained relief with warm soaks, 1 patient had massage only, and 2 patients were lost to follow up. Fourteen were examined in the emergency department, and 14 were treated at home. CONCLUSION: Although some injection injuries must be treated in an emergency facility, many can be treated at home. Immediate referral to a health care facility is not needed in all cases and at times is unwarranted. PMID- 11902700 TI - Legislators' views on tobacco policy: are there regional differences in Kentucky? AB - BACKGROUND: Tobacco-growing states have few tobacco control laws, which are relatively weak compared with those in non-tobacco-growing states. Region of the country has been shown to be a predictor of legislators' intentions to vote for cigarette tax increases. METHODS: A total of 116 lawmakers (84%) participated in face-to-face interviews before the 1998 Kentucky General Assembly. Five regions of Kentucky were identified by the five political caucuses. Legislative voting records on two tobacco control bills introduced during the 1998 Kentucky General Assembly were examined. RESULTS: There was little regional variation in opinions toward tobacco control policy among Kentucky legislators. Regional variation was evident only in relation to reducing the state's dependence on tobacco production, raising cigarette taxes, and adopting a law to prohibit teen possession of tobacco products. CONCLUSIONS: Health advocates from tobacco growing states might use regional information to garner support for selected tobacco control policies among lawmakers. PMID- 11902701 TI - Ketorolac for pain management after abdominal surgical procedures in infants. AB - BACKGROUND: While the use of intravenous ketorolac in infants less than 6 months old has not been previously reported, ketorolac may reduce opioid use and prevent opioid-associated side effects that are frequent in this age group. We reviewed our experience with ketorolac in infants to develop recommendations for using it after abdominal surgery. METHODS: We reviewed the records of 10 infants less than 6 months old who received ketorolac supplemented with morphine and 8 infants who received morphine alone for pain after abdominal surgery. RESULTS: Infants receiving ketorolac required less morphine than infants receiving morphine alone in the first 48 hours after surgery. Four patients receiving ketorolac did not require any supplemental morphine. CONCLUSIONS: Ketorolac reduces the amount of morphine required after abdominal surgery in infants less than 6 months old. The use of ketorolac deserves further study as a method of reducing opioid-associated adverse effects in this patient group. PMID- 11902702 TI - Medium and long-term growth in children receiving intranasal beclomethasone dipropionate: a clinical experience. AB - BACKGROUND: A 12-month controlled pediatric study of intranasal beclomethasone dipropionate (BDP) reported a 0.9 cm decrease in annual height growth velocity. Since children with allergic rhinitis may be treated for years, this report evaluates long-term height growth effects. METHODS: We reviewed the clinical charts of children with allergic rhinitis who were treated for the first time with intranasal BDP and were less than 10 years of age at initiation. Height was determined by stadiometry before intranasal corticosteroid therapy and compared with height at a final visit. RESULTS: Sixty children aged 24 to 117 months (mean age, 70 months) were treated for an average of 36 months. The pretherapy height percentile was 44.6, which increased to the 52.2 percentile at the final visit. CONCLUSIONS: Long-term clinical use of intranasal BDP in children was not associated with decreased height growth. This outcome may reflect decreased long term compliance compared with a short-term study. However, the treatment remained effective. Some children may be at special risk. Careful height measurements are recommended every 6 months. PMID- 11902703 TI - Patients with bipolar illness admitted to a general medical service. AB - BACKGROUND: This study was done to assess the size and characteristics of the patient population with bipolar illness treated on the general medical service of two divisions of the Charleston Area Medical Center, the largest hospital complex in West Virginia. METHODS: A total of 779 admitting and/or discharge summaries were reviewed. RESULTS: The average age of the manic patients (50.5 years) was lower than that of schizophrenic patients (56.1 years) or the general population (58.9 years). The most common reasons for admission in the manic group were chest pain (5 patients) and drug overdose (4 patients). Alcohol abuse was more common in the bipolar group (20%) than in the schizophrenic group (11%) or the general population (12%). CONCLUSIONS: Additional studies of the prevalence of bipolar disorder in West Virginia are warranted. PMID- 11902704 TI - Literary psychiatric observation and diagnosis through the ages: King Lear revisited. AB - Shakespeare's plays, and in particular King Lear, have been a favorite source of clinical observation and diagnosis for psychiatrists for the past two centuries. Most authors agree that the description of Lear's mental symptoms is remarkably consistent and close to life. This article summarizes previous attempts to diagnose the mental illness of King Lear, featuring, among others, such entities as mania, senile dementia, delirium, depression, and brief reactive psychosis, and offers a new diagnosis according to the modern diagnostic criteria, namely, bipolar I disorder, most recent episode manic, severe with psychotic features. PMID- 11902705 TI - Respiratory syncytial virus infection and the primary care physician. AB - Respiratory syncytial virus (RSV) infection is a common viral illness affecting almost all children within their first few years of life. In most young children, RSV results in a mild respiratory infection. It is, however, the single most important cause of bronchiolitis and pneumonitis in infancy and contributes to significant morbidity and even mortality in a subset of high-risk children. There are new developments in the diagnosis, treatment, and prevention of RSV infection in infants and children. Early recognition of young children at high risk for severe RSV infection and apnea can help to minimize the morbidity and mortality. PMID- 11902706 TI - Percutaneous balloon mitral valvuloplasty during a twin gestation. AB - Although percutaneous balloon mitral valvuloplasty is known to be a safe and effective procedure during a singleton pregnancy, its use during a twin gestation is limited to a single report with a poor obstetric outcome. In the case we are reporting, a 29-year-old nulliparous patient with a twin gestation was seen at 24 weeks' gestation with congestive heart failure caused by previously undiagnosed mitral stenosis. She had successful percutaneous balloon mitral valvuloplasty, and two healthy infants were subsequently delivered at 36 weeks' gestation. PMID- 11902707 TI - Diabetic gustatory sweating. AB - Gustatory sweating is a potential manifestation of autonomic dysfunction in diabetes. This syndrome is seen in long-standing diabetes and is associated with nephropathy, peripheral neuropathy, and other signs of dysautonomia. Symptoms of profuse head and neck diaphoresis with eating may suggest this clinical diagnosis. We present a patient who had complicated diabetes with symptoms of gustatory sweating and other evidence of dysautonomia. Diagnosis and treatment possibilities are discussed, with a review of the literature and an emphasis on bedside testing. PMID- 11902708 TI - Dermatofibrosarcoma protuberans with lung metastasis in a patient with progressive systemic sclerosis. AB - A female patient with progressive systemic sclerosis and pulmonary fibrosis had a dermatofibrosarcoma protuberans on the right thigh. After resection of the tumor, new lesions occurred in the scar, and wide excision was repeated. Two years later, a lung metastasis was discovered, and segmental resection was done. After 1 year of follow-up, no local recurrences or metastases were found. PMID- 11902709 TI - Airway obstruction due to bilateral giant pulmonary artery aneurysms. AB - Large pulmonary artery aneurysms are rare, but they are associated with significant morbidity and mortality. Significant airway obstruction due to extrinsic compression solely by a pulmonary artery aneurym is an extremely rare occurrence. We present a case of large bilateral pulmonary artery aneurysms causing extrinsic airway compression with collapse of the left primary bronchus in a 51-year-old woman. This is the first report in an adult in which airway compression due solely to the pulmonary artery aneurysm resulted in airway collapse. Furthermore, we describe the use of interventional bronchoscopy with stent placement as a minimally invasive alternative to surgery for treatment of these patients. PMID- 11902710 TI - Syncope in an adult with uncontrolled asthma. AB - Cough syncope occurs primarily in middle-aged male smokers with chronic obstructive pulmonary disease (COPD). It has also been described in children with asthma. I report the case of a 34-year-old nonsmoker who had syncope due to coughing, and who also related a 1-year history of cough with wheezing. Chest examination revealed diffuse wheezing and a prolonged expiratory phase, and pulmonary function testing with a methacholine challenge confirmed hyperreactive airways. Notably, while undergoing spirometric testing he had a recurrent syncopal episode. His syncope resolved with medical therapy for asthma. PMID- 11902711 TI - Importance of individualized decision in the management of massive pulmonary embolism. PMID- 11902712 TI - A "hospitalist" rotation increases short-term knowledge of fourth-year medical students. PMID- 11902713 TI - The Escherichia coli K-12 sheA gene encodes a 34-kDa secreted haemolysin. AB - Haemolytic toxins belong to one of several classes of virulence factors that contribute to bacterial pathogenicity. The non-pathogenic Escherichia coli K-12 laboratory strain was considered for years to be non-haemolytic. However, overproduction of several transcriptional regulators induced the appearance of a haemolytic activity that is absent under usual laboratory growth conditions. In this work, we have cloned and characterized an E. coli K-12 gene, sheA, whose overexpression results in a haemolytic phenotype. It maps to min 27 on the genetic map, and codes for a 34-kDa polypeptide with at least one putative transmembrane segment. This polypeptide, which has neither signal peptide nor other known secretory motifs, is secreted to the medium during the exponential growth phase. In vitro coupled transcription/translation assays, using a plasmid carrying only the sheA gene as template, resulted in the production of a polypeptide with haemolytic activity per se. Our results demonstrate that the sheA gene actually encodes the E. coli K-12 chromosomal haemolysin. The SheA haemolysin does not resemble other known cytolytic toxins, and it may represent the prototype of a novel family, as suggested by the presence of homologues in several E. coli pathogenic strains and in Shigella flexneri. PMID- 11902714 TI - PilC of pathogenic Neisseria is associated with the bacterial cell surface. AB - Adherence of pathogenic Neisseria to target host cells is mediated by pili. PilC1 and PilC2 are two high-molecular-weight proteins involved in pilus assembly and cellular adherence functions of the pili. Inactivation of pilC1 or pilC2 in N. meningitidis resulted in clones that expressed the same number of pili as the parent, contained no alterations in pilE and showed no detectable differences in PilE glycosylation. However, the PilC2+ pilC1- mutant showed much reduced adherence to target cells, indicating that production of PilC1 is essential for pilus-mediated adherence. To study further the functional differences between the meningococcal pilC genes, we determined the complete nucleotide sequence of pilC1 and pilC2 of N. meningitidis. Alignment of six PilC sequences demonstrated that PilC is composed of both conserved and variable regions. By immunogold labelling of bacterial sections we showed that PilC is present in the membranes of both piliated and non-piliated bacteria. Further, we demonstrated that PilC is associated with the bacterial cell surface. PMID- 11902715 TI - The Rhizobium meliloti exoK gene and prsD/prsE/exsH genes are components of independent degradative pathways which contribute to production of low-molecular weight succinoglycan. AB - When grown on medium supplemented with the succinoglycan-binding dye, Calcofluor, and visualized under UV light, colonies of Rhizobium meliloti (Sinorhizobium meliloti) exoK mutants produce a fluorescent halo with a delayed onset relative to wild-type colonies. By conducting transposon mutagenesis of exoK mutants of R. meliloti and screening for colonies with even more severe delays in production of these fluorescent halos, we identified three genes, designated prsD, prsE, and exsH, which are required for the eventual production of fluorescent halos by exoK colonies. Nucleotide sequence indicates that the prsD and prsE genes encode homologues of ABC transporters and membrane fusion proteins of Type I secretion systems, respectively, whereas exsH encodes a homologue of endo-1,3-1,4-beta glycanases with glycine-rich nonameric repeats typical of proteins secreted by Type I secretion systems. The exoK gene and the prsD/prsE/exsH genes were shown to be components of independent pathways for production of extracellular succinoglycan degrading activities and for production of low-molecular-weight succinoglycan by R. meliloti. Based on these results, we propose that ExsH is a succinoglycan depolymerase secreted by a Type I secretion system composed of PrsD and PrsE, and that the ExsH and ExoK glycanases contribute to production of low molecular-weight succinoglycan. PMID- 11902716 TI - The Rhizobium leguminosarum prsDE genes are required for secretion of several proteins, some of which influence nodulation, symbiotic nitrogen fixation and exopolysaccharide modification. AB - NodO is a secreted protein from Rhizobium leguminosarum bv. viciae with a role in signalling during legume nodulation. A Tn5-induced mutant was identified that was defective in NodO secretion. As predicted, the secretion defect decreased pea and vetch nodulation but only when the nodE gene was also mutated. This confirms earlier observations that NodO plays a particularly important role in nodulation when Nod factors carrying C18:1 (but not C18:4) acyl groups are the primary signalling molecules. In addition to NodO secretion and nodulation, the secretion mutant had a number of other characteristics. Several additional proteins including at least three Ca2+-binding proteins were not secreted by the mutant and this is thought to have caused the pleiotropic phenotype. The nodules formed by the secretion mutant were unable to fix nitrogen efficiently; this was not due to a defect in invasion because the nodule structures appeared normal and nodule cells contained many bacteroids. The mutant formed sticky colonies and viscous liquid cultures; analysis of the acidic exopolysaccharide revealed a decrease in the ratio of reducing sugars to total sugar content, indicating a longer chain length. The use of a plate assay showed that the mutant was defective in an extracellular glycanase activity. DNA sequencing identified the prsDE genes, which are homologous to genes encoding protease export systems in Erwinia chrysanthemi and Pseudomonas aeruginosa. An endoglycanase (Egl) from Azorhizobium caulinodans may be secreted from R. leguminosarum bv. viciae in a prsD-dependent manner. We conclude that the prsDE genes encode a Type I secretion complex that is required for the secretion of NodO, a glycanase and probably a number of other proteins, at least one of which is necessary for symbiotic nitrogen fixation. PMID- 11902717 TI - Residence at the expression site is necessary and sufficient for the transcription of surface antigen genes of Pneumocystis carinii. AB - The major surface glycoprotein (MSG) of P. carinii f. sp. carinii is a family of proteins encoded by a family of heterogeneous genes. Messenger RNAs encoding different MSG isoforms start with the same sequence, called the upstream conserved sequence (UCS), which is encoded by a single locus. The mechanism by which the UCS becomes part of different MSG mRNAs is not obvious because at least 15 loci, which are distributed throughout the genome, encode MSGs. One possibility is that attachment to the UCS locus is required for the transcription of an MSG gene. The alternative to this expression site model is that mRNAs acquire the UCS by RNA splicing. To distinguish between these two models, UCS/MSG junctions in the genome were compared with UCS/MSG junctions in mRNA. The UCS/MSG junctions in the mRNA matched those in the genome, as would be expected if splicing did not contribute to the attachment of the UCS to the 5' ends of MSG mRNAs. Given that few if any MSG mRNAs lack the UCS, the correspondence between the UCS/MSG junctions in transcripts and those in the genome indicates that attachment to the UCS is both necessary and sufficient for transcription of an MSG gene. PMID- 11902718 TI - Unsuspected prophage-like elements in Salmonella typhimurium. AB - We present evidence for the existence of two large (approximately 50 kb) excisable segments in the chromosome of Salmonella typhimurium. The two elements- designated Gifsy-1 and Gifsy-2--cover, respectively, the 57 units and the 24 units of the genetic map where they contribute indicative rare restriction sites. The two elements are closely interrelated and both contain a region of sequence similarity to the recE locus of the Rac prophage of Escherichia coli. Mutations within this region of Gifsy-1 yield the classical 'Sbc' phenotype: they suppress the recombination defect of recB mutants, apparently by activating a normally silent recE-like gene. At the same time, these 'sbcE' mutations activate a Xis type function that promotes excision of one or other of the two elements. Predictably, curing of Gifsy-1 results in the loss of recB mutant suppression. Surprisingly, the suppressor phenotype is also lost in cells cured for Gifsy-2 even though the Gifsy-1-associated sbcE mutation is still present. Moreover, the excision frequency of Gifsy-1 drops dramatically in Gifsy-2-cured cells. Thus, both elements must co-operate in the activation of recombination and excision functions. Overall, the data presented here suggest that Gifsy-1 and Gifsy-2 are cryptic prophages. They are distinct from previously described Fels prophages. Unlike Fels, they are not specific to S. typhimurium strain LT2 since they are both also found in a virulent S. typhimurium isolate (ATCC 14028s). PMID- 11902720 TI - Requirements for ribosomal protein S1 for translation initiation of mRNAs with and without a 5' leader sequence. AB - It has previously been proposed that Escherichia coli ribosomal protein S1 is required for the translation of highly structured mRNAs. In this study, we have examined the influence of structural features at or near the start codon of different mRNAs. The requirement for ribosomal protein S1 for translation initiation was determined when (i) the ribosome-binding site (RBS) was either preceded by a 5' non-translated leader sequence; (ii) the RBS was located 5' proximal to a mRNA start codon; and (iii) the start codon was the 5' terminal codon as exemplified by leaderless mRNAs. In vitro translation studies revealed that the leaderless lambda cl mRNA is translated with Bacillus stearothermophilusribosomes, naturally lacking a ribosomal protein S1 homologue, whereas ompA mRNA containing a 5' leader is not. These studies have been verified by toeprinting with E. coli ribosomes depleted for S1. We have shown that S1 is required for ternary complex formation on ompA mRNA but not for leaderless mRNAs or for mRNAs in which the RBS is close to the 5' end. PMID- 11902719 TI - Osmostress response in Bacillus subtilis: characterization of a proline uptake system (OpuE) regulated by high osmolarity and the alternative transcription factor sigma B. AB - Exogenously provided proline has been shown to serve as an osmoprotectant in Bacillus subtilis. Uptake of proline is under osmotic control and functions independently of the known transport systems for the osmoprotectant glycine betaine. We cloned the structural gene (opuE) for this proline transport system and constructed a chromosomal opuE mutant by marker replacement. The resulting B. subtilis strain was entirely deficient in osmoregulated proline transport activity and was no longer protected by exogenously provided proline, attesting to the central importance of OpuE for proline uptake in high-osmolarity environments. The transport characteristics and growth properties of the opuE mutant revealed the presence of a second proline transport activity in B. subtilis. DNA sequence analysis of the opuE region showed that the OpuE transporter (492 residues) consists of a single integral membrane protein. Database searches indicated that OpuE is a member of the sodium/solute symporter family, comprising proteins from both prokaryotes and eukaryotes that obligatorily couple substrate uptake to Na+ symport. The highest similarity was detected to the PutP proline permeases, which are used in Escherichia coli, Salmonella typhimurium and Staphylococcus aureus for the acquisition of proline as a carbon and nitrogen source, but not for osmoprotective purposes. An elevation of the osmolarity of the growth medium by either ionic or non-ionic osmolytes resulted in a strong increase in the OpuE-mediated proline uptake. This osmoregulated proline transport activity was entirely dependent on de novo protein synthesis, suggesting a transcriptional control mechanism. Primer extension analysis revealed the presence of two osmoregulated and tightly spaced opuE promoters. The activity of one of these promoters was dependent on sigma A and the second promoter was controlled by the general stress transcription factor sigma B. PMID- 11902721 TI - Association of a partial H-rpt element with the type 3 capsule locus of Streptococcus pneumoniae. PMID- 11902722 TI - Disrespectful type IV pilins. PMID- 11902723 TI - Negative autoregulation of the Rhizobium meliloti fixK gene is indirect and requires a newly identified regulator, FixT. AB - fixK genes are crp/fnr homologues that have been discovered in diverse Rhizobium spp., in which they are usually essential for symbiotic nitrogen fixation. One recurrent function of fixK genes in rhizobia is to activate the transcription of operons required for respiration in the microoxic environment of the nodule. In a similar manner to its Escherichia coli crp and fnr homologues, R. meliloti fixK regulates its own expression negatively. However, we demonstrate here that fixK negative autoregulation is not direct and, instead, involves a newly identified gene, fixT, the expression of which depends on fixK. Inactivation of fixT resulted in derepression of fixK expression under free-living microoxic conditions. Furthermore, constitutively expressed fixT strongly repressed fixK lacZ expression in the absence of a functional fixK gene. Several lines of evidence indicate that fixT is active via its protein product FixT. FixT does not resemble any protein present in databases so far. Nodules induced by a fixT mutant were Fix+, thus demonstrating that fixT is not essential for symbiotic nitrogen fixation. PMID- 11902724 TI - Enzymic and genetic basis for bacterial growth on malonate. AB - Various bacteria are able to grow aerobically or anaerobically on malonate as sole source of carbon and energy. Independent of the mechanism for energy conservation, the decarboxylation of malonate is the key reaction in the decomposition of this compound. To achieve malonate decarboxylation under physiological conditions, the substrate must be converted into an activated (thioester) derivative. We report here on the malonate decarboxylases of Malonomonas rubra and Klebsiella pneumoniae. These enzymes perform an interesting substrate activation mechanism by generating a malonyl thioester with the enzyme. Formation of the malonyl-S-enzyme involves an 'activation module' that comprises the acetylation of a specific thiol group of an acyl carrier protein (ACP) and the transfer of the ACP moiety to malonate, yielding malonyl-S-ACP and acetate. The malonyl-S-ACP is subsequently decarboxylated with regeneration of the acetyl ACP. The malonate activation mechanism is related to the activation of citrate by citrate lyase. The relationship extends to the identical 2'-(5''-phosphoribosyl) 3'-dephospho-CoA thiol cofactor that is bound covalently to the corresponding ACP subunit. In Klebsiella pneumoniae, malonate is decarboxylated by a water-soluble enzyme complex. In the anaerobic bacterium Malonomonas rubra, malonate decarboxylation is catalysed by a set of water-soluble as well as membrane-bound enzymes that function together in converting the free energy of the decarboxylation reaction into delta muNa+. Therefore, this malonate decarboxylase includes a biotin carrier protein that accepts the CO2 moiety from malonyl-S-ACP and delivers it to a membrane-bound decarboxylase acting as a Na+ pump. Genes encoding the individual protein components that perform the decarboxylation of malonate in K. pneumoniae or M. rubra have been identified within the mdc and mad gene clusters respectively. The function of most of the derived proteins could be envisaged from sequence similarities with proteins of known functions. The genetic evidence firmly supports the idea that malonate decarboxylation is carried out by the two different decarboxylases, as deduced from the biochemical studies of the enzymes. PMID- 11902725 TI - Alterations in the flow of one-carbon units affect KinB-dependent sporulation in Bacillus subtilis. AB - Endospore formation in Bacillus subtilis is primarily dependent on the phosphorylation of the key transcription factor Spo0A by two major kinases, KinA and KinB, thought to be activated by distinct signals. Using a strategy designed to detect mutations that specifically affect the signalling pathway to KinB, we have isolated a Tn10 insertion mutant in one of two adjacent lrp-like genes coding for homologues of the Escherichia coli leucine-responsive regulatory protein (Lrp) and another mutant in the glyA gene encoding the serine hydroxymethyl transferase (SHMT). SHMT catalyses interconversion of serine and glycine while transferring the resulting one-carbon unit into the C1 pool through methylene tetrahydrofolate. Sporulation experiments performed in a series of supplemented media indicated that the role of SHMT in the KinB pathway is to feed the pool of C1 units recruited for the biosynthesis of key metabolites, which include the methyl donor S-adenosyl-methionine (SAM). The results of experiments using L-ethionine suggest that SAM is involved in post-synthetic methylation reactions or biosynthesis of metabolites that serve to activate KinB. Truncated LrpA and LrpB peptides that have retained the DNA-binding domain but have lost the C-terminal half of the protein appear to act as repressors of glyA transcription and KinB-dependent sporulation. However, deletions of lrpA, lrpB or lrpAB have little effect on glyA transcription or sporulation through KinB, suggesting that other effectors, such as additional Lrp homologues, may act in conjunction with LrpA and LrpB. Our results indicate that lrpA-lrpB together with the biosynthetic glyA gene lie on a common signalling pathway meant to activate the KinB sensor kinase. PMID- 11902726 TI - Nascent membrane and presecretory proteins synthesized in Escherichia coli associate with signal recognition particle and trigger factor. AB - The Escherichia coli signal recognition particle (SRP) and trigger factor are cytoplasmic factors that interact with short nascent polypeptides of presecretory and membrane proteins produced in a heterologous in vitro translation system. In this study, we use an E. coli in vitro translation system in combination with bifunctional cross-linking reagents to investigate these interactions in more detail in a homologous environment. Using this approach, the direct interaction of SRP with nascent polypeptides that expose particularly hydrophobic targeting signals is demonstrated, suggesting that inner membrane proteins are the primary physiological substrate of the E. coli SRP. Evidence is presented that the overproduction of proteins that expose hydrophobic polypeptide stretches, titrates SRP. In addition, trigger factor is efficiently cross-linked to nascent polypeptides of different length and nature, some as short as 57 amino acid residues, indicating that it is positioned near the nascent chain exit site on the E. coli ribosome. PMID- 11902727 TI - Induction of the Bacillus subtilis ptsGHI operon by glucose is controlled by a novel antiterminator, GlcT. AB - Glucose is the preferred carbon and energy source of Bacillus subtilis. It is transported into the cell by the glucose-specific phosphoenolpyruvate:sugar phosphotransferase system (PTS) encoded by the ptsGHI locus. We show here that these three genes (ptsG, ptsH, and ptsI) form an operon, the expression of which is inducible by glucose. In addition, ptsH and ptsl form a constitutive ptsHI operon. The promoter of the ptsGHI operon was mapped and expression from this promoter was found to be constitutive. Deletion mapping of the promoter region revealed the presence of a transcriptional terminator as a regulatory element between the promoter and coding region of the ptsG gene. Mutations within the ptsG gene were characterized and their consequences on the expression of ptsG studied. The results suggest that expression of the ptsGHI operon is subject to negative autoregulation by the glucose permease, which is the ptsG gene product. A regulatory gene located upstream of the ptsGHI operon, termed glcT, was also identified. The GlcT protein is a novel member of the BglG family of transcriptional antiterminators and is essential for the expression of the ptsGHI operon. A deletion of the terminator alleviates the need for GlcT. The activity of GlcT is negatively regulated by the glucose permease. PMID- 11902729 TI - Mutations leading to increased levels of resistance to glycopeptide antibiotics in VanB-type enterococci. AB - The vanB gene cluster mediates glycopeptide resistance by production of peptidoglycan precursors ending in the depsipeptide D-alanyl-D-lactate (D-Ala-D Lac) instead of D-Ala-D-Ala found in susceptible enterococci. Synthesis of D-Ala D-Lac and hydrolysis of D-Ala-D-Ala is controlled by the VanR(B)S(B) two component regulatory system that activates transcription of the resistance genes in response to vancomycin but not to teicoplanin. Two substitutions (A3C-->G or D168-->Y) in the VanS(B) sensor kinase resulted in induction by teicoplanin, indicating that the N-terminal domain of the protein was involved in glycopeptide sensing. A substitution (T237-->K) located in the vicinity of the putative autophosphorylation site of VanS(B) (H233) was associated with a constitutive phenotype and affected a conserved residue known to be critical for the phosphatase activity of related kinases. A mutant producing an impaired host D Ala:D-Ala ligase required vancomycin for growth, since D-Ala-D-Lac was only produced under inducing conditions. The ddl and vanS(B) mutations, alone or in combination, resulted in various resistance phenotypes that were determined by the amount of D-Ala-D-Ala and D-Ala-D-Lac incorporated into peptidoglycan precursors under different inducing conditions. PMID- 11902728 TI - Molecular organization of the genes required for the synthesis of type 1 capsular polysaccharide of Streptococcus pneumoniae: formation of binary encapsulated pneumococci and identification of cryptic dTDP-rhamnose biosynthesis genes. AB - We report here the molecular organization of the capsular locus (cap1) of the type 1 pneumococcus. This locus is located between dexB and aliA and flanked by IS1167 insertion elements. Sequence analysis showed that the cluster contains 11 genes (cap1A to cap1K), which are apparently arranged as a single transcriptional unit. The presence of a functional promoter (cap1p) located upstream of cap1A has been demonstrated and the transcription start point was mapped by primer extension analysis. A 14.3 kb fragment containing the genes cap1ABCDEFGHIJK and including cap1p was sufficient to allow the synthesis of a type 1 capsule in Streptococcus pneumoniae. An internal deletion of cap1E leads to an unencapsulated phenotype demonstrating that this gene is essential for capsular production. The cap1K gene has been expressed in Escherichia coli resulting in UDP-glucose dehydrogenase (UDP-GlcDH) activity. Moreover, this gene was able to restore the synthesis of type 3 capsule when cloned into a plasmid and introduced by transformation into S. pneumoniae cap3A mutants deficient in UDP-GlcDH. In marked contrast with what was previously thought, recombination between cap1K and cap3A does occur. We provide data on the molecular mechanism that leads to the formation of binary encapsulated pneumococcal cells, i.e. strains that simultaneously produce type 1 and type 3 capsules. Downstream of cap1K, one truncated and three complete open reading frames homologous to those involved in the biosynthesis of dTDP-rhamnose, a monosaccharide that does not participate in the formation of type 1 polysaccharide, have been identified in all the clinical strains of type 1 pneumococcus tested. Our results provide new insights into the generation of capsule diversity in pneumococci. PMID- 11902730 TI - Down syndrome, transient myeloproliferative syndrome, and leukemia: bridging development and neoplasia. PMID- 11902731 TI - Telomerase and the benign and malignant megakaryoblastic leukemias of Down syndrome. AB - The most common form of leukemia in Down syndrome patients is megakaryoblastic leukemia. There are two forms of the disease. Transient leukemia (TL) is a form of megakaryoblastic leukemia that occurs in newborns with Down syndrome and usually disappears spontaneously within the first 3 months of life. Acute megakaryoblastic leukemia (AMKL) occurs in Down syndrome children within the first 4 years of life and is fatal without treatment. The megakaryoblasts of TL and AMKL are indistinguishable by light and electron microscopy; yet, TL is benign and AMKL is malignant. One of the hallmarks of many malignancies is the expression of telomerase. It is therefore hypothesized that the transient, benign form of megakaryoblastic leukemia (TL) would not contain telomerase activity, whereas telomerase would be demonstrable in the malignant form of the disease. Telomerase activity was determined in the blood and/or bone marrow aspirates in 29 cases of AMKL and 34 cases of TL. The authors found telomerase activity in 15 of 29 (52%) cases, of AMKL and in only 4 of 34 (12%) cases of TL (P < 0.001). Furthermore, three of the four telomerase-positive TL cases were particularly severe, of which two were fatal. Telomerase activity is found frequently in the leukemic cells of the malignant form of megakaryoblastic leukemia but rarely in the benign form of the disease (TL). Observations provide evidence that telomerase may be a critical factor for the malignant conversion of leukemic cells. PMID- 11902732 TI - Alpha-interferon in combination with cytarabine in children with Philadelphia chromosome-positive chronic myeloid leukemia. AB - BACKGROUND: Philadelphia chromosome-positive chronic myelogenous leukemia (CML) is a rare disease in children, and the optimal therapy is not clearly defined in these patients when a human leukocyte antigen-identical donor is not available. The present work focuses on the therapeutic efficacy and the toxicity of interferon (IFN) alpha 2b in combination with cytosine arabinosine (Ara-C) in patients younger than age 18 years enrolled in the randomized trial CML 91, which compared the efficacy of IFN and cytosine arabinoside (Ara-C) with IFN alone in 810 patients with CML in the chronic phase. PATIENTS AND METHODS: Twelve patients younger than age 18 years were enrolled in the randomized trial CML 91. Hydroxyurea and IFN (5 million units/m2, once a day) were given as initial treatment in all patients. After randomization, six patients received IFN (5 million units/m2, once per day) and Ara-C (20 mg/m2 for 10 days each month) (IFN plus Ara-C group), and six patients received IFN alone (5 million units/m2 once per day) (IFN group). RESULTS: Six months after the beginning of the treatment, a complete hematologic response was obtained in all the patients in the IFN plus Ara-C group and in four patients in the IFN group. A major cytogenetic response was observed in three patients in the IFN plus Ara-C group and in two patients in the IFN group. Five patients from the IFN group who crossed over to receive Ara-C did not experience additional hematologic toxicity. Three patients in the IFN plus Ara-C group and two from the IFN group are alive, in major cytogenetic response, with a follow-up of 18 to 48 months. CONCLUSION: The combination of IFN and Ara-C induces complete hematologic and major cytogenetic responses and is well tolerated in patients younger than age 18 years with CML. This combination may offer an alternative to bone marrow transplantation in children in the chronic phase of CML without a histocompatible donor. PMID- 11902733 TI - Transient myeloproliferative disorder, a disorder with too few data and many unanswered questions: does it contain an important piece of the puzzle to understanding hematopoiesis and acute myelogenous leukemia? PMID- 11902734 TI - Predictors of outcome in the pediatric intensive care units of children with malignancies. AB - PURPOSE: Children with malignancies in whom life-threatening complications develop are traditionally considered as having a grim prognosis. Clinical predictors of short-term outcome for rational triage to pediatric intensive care units (PICU) were retrospectively assessed. PATIENTS AND METHODS: The records of 94 children consecutively admitted to the PICU at the authors' institution between January 1989 and January 1999 were reviewed, and predictors of 30-day mortality rates were delineated using stepwise logistic regression. RESULTS: The children's mean age was 7.3 years (range, 2-21). Their diseases included hematologic malignancies 45 (48%), extracranial solid tumors 21 (22%), and intracranial tumors 28 (30%). The overall 30-day survival rate was 66%. Mortality was highest among children admitted for respiratory failure (40%). High mortality was also found for those with circulatory collapse (33.3%) and neurologic deterioration (31%). The admitting pediatric risk of mortality score (PRISM) among the survivors was 6.6 +/- 1.3, compared with 15.2 +/- 3 among nonsurvivors (P < 0.01). The number of organ system failures was higher among the nonsurvivors on admission (P < 0.001). The need for ventilatory or inotropic support corresponded to worse outcome (P < 0.001 or P < 0.01, respectively). Overall, 36 (38%) of the children had sepsis during their PICU stay, with a mortality rate of 50% compared with 24% among nonseptic children (P < 0.01). Sepsis present on admission was later correlated with the development of organ system failure (P < 0.01). CONCLUSIONS: New trends in therapeutic approaches to children with malignancies can clearly improve outcome. The high (66%) survival rate justifies policy of early admission to the PICU of children in whom signs of multiorgan involvement start to develop, as reflected by high PRISM and the need for ventilatory or inotropic support. Further refinement of reliable clinical predictors of survival will enable better triage of these children to the PICU for possible prevention of systemic complications and reduction of mortality rates. PMID- 11902735 TI - High-grade osteosarcoma of the extremity: differences between localized and metastatic tumors at presentation. AB - BACKGROUND: In osteosarcoma, as in other tumors, the presence of metastases at presentation is generally considered a consequence of late diagnosis. To verify this, the authors investigated whether there was a relationship between the stage of the disease at presentation and several clinical and pathologic characteristics, including the interval between the onset of first symptoms or signs and the final diagnosis. PATIENTS AND METHODS: One thousand seventy-one patients with high-grade osteosarcoma of the extremity were observed between 1980 and 1999. Of these, 891 had a localized tumor and 180 had metastases at the time of diagnosis. RESULTS: Compared with patients with localized disease, patients with detectable metastases at the time of diagnosis had higher serum levels of alkaline phosphatase, larger primary lesions, and tumors often located in the femur and humerus. In terms of time to diagnosis, the interval between the onset of first symptoms and the final diagnosis was significantly shorter in patients with metastases than in patients with localized tumor. This surprising finding probably reflects a more rapid growth of the tumor. CONCLUSIONS: These results suggest a different biologic phenotype and aggressiveness of the tumor in a subgroup of patients and that the stage of the disease at presentation depends more on the properties of these tumors than on late diagnosis. PMID- 11902736 TI - Response without shrinkage in bilateral Wilms tumor: significance of rhabdomyomatous histology. AB - PURPOSE: To test the hypothesis that poor response to chemotherapy in patients with bilateral Wilms tumor may be associated with the appearance of rhabdomyomatous histology, suggesting a differentiation response. METHODS: Twenty six patients with bilateral Wilms tumor were treated at the authors' hospital between 1985 and 1995. Radiologic response to presurgical chemotherapy was assessed, and postsurgery histology was reviewed. RESULTS: There was a significant association between rhabdomyomatous differentiation in postchemotherapy surgical specimens and poor radiologic response. Poor response did not, however, necessarily mean poor outcome: of 11 patients with rhabdomyomatous differentiation, 7 are alive and disease-free, 2 died of complications, and only 2 died of uncontrolled Wilms tumor. CONCLUSIONS: Rhabdomyomatous differentiation in postchemotherapy bilateral Wilms tumor is associated with poor radiologic response. This observation may indicate a differentiation response rather than an absolute failure of response to chemotherapy. Clinical measures other than tumor volume are needed to distinguish between tumors that respond to chemotherapy but do not shrink, and those that genuinely do not respond. PMID- 11902738 TI - Evaluation of risk prediction criteria for episodes of febrile neutropenia in children with cancer. AB - PURPOSE: To evaluate the feasibility of risk stratification of children with cancer and febrile neutropenia using a simple set of criteria from data available to the clinician at the time of the patient's presentation. PATIENTS AND METHODS: This study is a retrospective cohort study of all children with cancer admitted to a single institution with fever and neutropenia (defined as an absolute neutrophil count < 500 cells/mm3) in a 1-year period. Patients were defined a priori as low risk if they were outpatients at the time of presentation with febrile neutropenia, had an anticipated duration of neutropenia less than 7 days, and had no significant comorbidity. All others were considered high risk. Data was analyzed by first admission for each patient and secondarily for all admissions for febrile neutropenia. RESULTS: There were 188 admissions in 104 patients for febrile neutropenia during the study period. Of these 47% were high risk and 53% were low risk. The duration of fever was not significantly different in the two groups. However, the duration of neutropenia and the length of hospital stay were significantly longer in the high-risk group. The frequency of bacteremia, other documented infection, and serious medical complications was significantly different in the two groups. Overall, the rate of any adverse event was 4% in the low-risk group versus 41% in the high-risk group. CONCLUSIONS: Simple criteria available to the clinician at the time of evaluation of the child with cancer who has fever and neutropenia allow the selection of a population at low risk for bacteremia or serious medical complication. A prospective study is planned using these risk criteria, evaluating outpatient oral antibiotic therapy in low-risk children with cancer. PMID- 11902737 TI - Incidence of anemia in children with solid tumors or Hodgkin disease. AB - Anemia is a hematologic abnormality commonly discussed during the treatment of childhood cancer, but its incidence has not been previously reported. As the basis for determining the incidence of anemia, this retrospective review of medical records combined databases containing the records of all patients 1 to 18 years of age with newly diagnosed neuroblastoma, rhabdomyosarcoma, Hodgkin disease, Ewing sarcoma, or osteosarcoma from two pediatric oncology centers. Data from 405 patients were included in the analysis of hemoglobin at the time of diagnosis. Across diagnoses, 51% to 74% of patients were anemic using the Centers for Disease Control and Prevention age- and sex-specific values to define anemia. The long-term complications of anemia in children with cancer are unknown. Further investigation of the clinical significance of anemia, including its impact on quality of life, is warranted. PMID- 11902739 TI - Transfusion volume in infants with very low birth weight: a randomized trial of 10 versus 20 ml/kg. AB - BACKGROUND: Although preterm infants often require transfusions of red blood cells for anemia of prematurity, the optimal volume of blood to be transfused has not been established. OBSERVATIONS: Infants with birth weights between 500 and 1,500 g were randomly assigned to receive 10 or 20 mL/kg red blood cells. Infants with transfusions of 20 mL/kg had a greater hemoglobin (14.2 +/- 1.9 vs. 12.0 +/- 1.9 g/dL, P = 0. 003) and hematocrit (41.2 +/- 5.9 vs. 32.3 +/- 7.1%, P = 0.001) levels after transfusion compared with those who received transfusions of 10 mL/kg. There were no measured differences in pulmonary function in either group after transfusion. CONCLUSIONS: Transfusion with 20 mL/kg red blood cells produces a significantly greater increase in hemoglobin and hematocrit levels than does a transfusion with 10 mL/kg, without any detrimental effects on pulmonary function. PMID- 11902740 TI - Clinical and laboratory features of 178 children with recurrent epistaxis. AB - PURPOSE: To determine the clinical and laboratory features of 178 children referred for the evaluation of recurrent epistaxis to an outpatient hematology clinic in a university medical center. PATIENTS AND METHODS: Medical records of 3681 outpatient pediatric hematology referrals were retrospectively review, and 178 children with recurrent epistaxis from 1985 to 1999 were identified. Historic (other bleeding symptoms: gingival bleeding, easy bruising, menorrhagia, and gross blood in the urine or stool: duration and severity of the epistaxis episodes; and family history of bleeding) and laboratory (complete blood count and coagulation tests) data were analyzed. RESULTS: There were 103 boys and 75 girls with a median age of 84 months (range 15-219 months). Sixty-seven percent (n = 119) did not have a coagulopathy diagnosed and 33% (n = 59) did. The diagnoses included von Willebrand disease in 33, platelet aggregation disorders in 10, thrombocytopenia in seven, mild factor VIII deficiency in three, Bernard Soulier syndrome in two, factor VII deficiency in one, factor IX deficiency in one, and factor XI deficiency in one, and coagulation inhibitor in one. Of the historic data, only a family history of bleeding was predictive of diagnosing a coagulopathy (P = 0.023). The duration and severity of the epistaxis and the presence of other bleeding symptoms had no predictive value. Children with a coagulopathy diagnosed had a longer median partial thromboplastin time (PTT) (33.1 vs. 30.5 seconds; P = 0.012). CONCLUSIONS: One-third of children presenting with recurrent epistaxis have a diagnosable coagulopathy. A positive family history and a prolonged PPT are useful predictive data. PMID- 11902741 TI - Acute megakaryoblastic leukemia after transient myeloproliferative disorder with clonal karyotype evolution in a phenotypically normal neonate. AB - We report a case of transient myeloproliferative disorder (TMD) in a neonate without features of Down syndrome (DS) with clonal karyotype evolution, after apparent spontaneous resolution of TMD, but eventually progressing to acute megakaryoblastic leukemia (AMKL). The patient had petechiae, thrombocytopenia, and blastemia. Trisomy 21 with a satellited Y chromosome (Yqs) was found in proliferating blasts. A stimulated peripheral blood culture confirmed the constitutional origin of the Yqs, but did not reveal the presence of any trisomic 21 cell. By the age of 3 months, clonal chromosome evolution in the form of an interstitial deletion of the long-arm of chromosome 13 [del(13)(q13q31)] was detected along with trisomy 21 in unstimulated bone marrow cultures. However, remission was achieved without treatment at the age of 4 months. Trisomy 21 and del(13)(q13q31) were not identified in either cytogenetics or fluorescence in situ hybridization studies at that time. The child was asymptomatic until the age of 20 months when anemia and thrombocytopenia prompted a bone marrow biopsy, revealing changes consistent with AMKL. The remission proceeded by clonal karyotype evolution in a neonate with TMD demonstrates that clonal karyotype evolution does not indicate an immediately progressive disease. However, the development of AMKL after TMD in this case illustrates the increased risk for leukemia in TMD cases, even without DS. The gradual clonal evolution of the blasts in our patient suggests that "multiple hits" oncogenesis applies to TMD progression to acute leukemia. PMID- 11902742 TI - Presentation of M4 acute myeloid leukemia in anuric renal failure with hyperuricemia and enlarged kidneys. AB - Extramedullary acute myeloid leukemia (AML) is not uncommon. It has been shown to involve the kidneys in most postmortem cases but is most often clinically insignificant. By contrast, acute tumor lysis syndrome is rare in AML, especially at initial diagnosis. The authors report the management of a patient with AML who had acute tumor lysis syndrome that was probably potentiated by renal leukemia and resulted in renal failure. This patient achieved remission with dose-modified induction chemotherapy administered while he was dialysis-dependent. PMID- 11902743 TI - Use of 2-chlorodeoxyadenosine to treat infantile myofibromatosis. AB - A 3-year-old boy had fever and bone pain. Magnetic resonance imaging of his femurs showed marrow replacement; iliac crest marrow biopsy revealed myelofibrosis. Although the pathologic criteria for Langerhans cell histiocytosis were not met, the clinical picture led to treatment with etoposide and methylprednisolone, without clinical improvement. One month after presentation, generalized tonic-clonic seizures occurred, and magnetic resonance imaging revealed parenchymal brain lesions. 2-chlorodeoxyadenosine was used. Because of the unexpected lack of response to etoposide and methylprednisolone, a second bone biopsy was performed. The diagnosis was revised to infantile myofibromatosis. After six courses of 2-chlorodeoxyadenosine, brain and bone lesions regressed, with resolution of the clinical symptoms. PMID- 11902744 TI - Down syndrome and the transient myeloproliferative disorder: why is it transient? PMID- 11902745 TI - Expansion of trisomy 8 and Sweet syndrome in a prolonged course of aplastic anemia. AB - We describe a 17-year-old boy with aplastic anemia who had Sweet syndrome develop with increasing expansion of trisomy 8. The diagnosis of aplastic anemia was made at 6 years of age. Cytopenias partially responded to danazol therapy. Cytogenetic studies of bone marrow (BM) cells were normal until the detection of trisomy 8 at 14 years of age. This clone increased with the progression of cytopenias. Cell sorting and fluorescence in situ hybridization analysis revealed that trisomy 8 was present only in nonlymphoid elements. When the patient was 17 years of age, Sweet syndrome developed. BM study showed myelodysplastic features, in which trisomy 8 occupied 74% of BM cells with additional chromosomal changes. Trisomy 8 may contribute to the late transformation of myeloid lineages in BM failure. PMID- 11902746 TI - Persistent developmental delay despite successful bone marrow transplantation for purine nucleoside phosphorylase deficiency. AB - A 10-month-old girl with a history of recurrent candidiasis, developmental delay, and a fulminant varicella infection is described. The diagnosis of purine nucleoside phosphorylase (PNP) deficiency was suggested by a reduced level of serum uric acid and confirmed by measurement of PNP activity. A human leukocyte antigen-matched bone marrow transplantation resulted in immune reconstitution, but poor neurodevelopmental progression. PMID- 11902747 TI - Sensitivity of a blood culture drawn through a single lumen of a multilumen, long term, indwelling, central venous catheter in pediatric oncology patients. AB - PURPOSE: The study assessed the sensitivity of a blood culture drawn from only one lumen of a multilumen, long-term, indwelling, central venous catheter in pediatric oncology patients. METHODS: Episodes of positive blood cultures were included if cultures were sent simultaneously from multiple lumens of a long term, indwelling, central line and if the culture was thought to represent a true bacteremia. RESULTS: Discordant results from cultures drawn simultaneously from different lumens occurred in 13 (32%) of 41 episodes. The estimated sensitivity of a culture drawn from a single lumen was 84%. CONCLUSION: Drawing blood cultures from all lumens of these lines should be considered in this patient population. PMID- 11902748 TI - Salmonella infection associated with a pet lizard in siblings with sickle cell anemia: an avoidable risk. PMID- 11902749 TI - Severe lung fibrosis after chemotherapy in a child with ataxia-telangiectasia. PMID- 11902750 TI - Successful engraftment of unrelated donor stem cells in two children with congenital amegakaryocytic thrombocytopenia. PMID- 11902751 TI - Incidence and treatment of potentially lethal diseases in transient leukemia of Down syndrome: Pediatric Oncology Group Study. AB - Transient leukemia (TL or transient myeloproliferative disorder) occurs in approximately 10% of newborn infants with Down syndrome. The disorder is characterized by the presence of megakaryoblasts in the peripheral blood; most cases resolve spontaneously within the first 3 months of life, and the child is well thereafter. However, there are cases in which a severe, potentially lethal form of disease develops, manifesting as hepatic fibrosis or cardiopulmonary failure. Hitherto, the incidence of these severe forms of the disease has not been reported. A prospective study of TL was conducted by the Pediatric Oncology Group (POG Study 9481) in which 48 children with TL were identified. Life threatening disease occurred in nine patients (19%); seven had hepatic fibrosis and two had cardiopulmonary failure. Five children died of the disease within the first 3 months of life, none of whom received antileukemic therapy. One patient died on day 31 after receiving minimal therapy within 1 day of death. Three children received low-dose cytosine arabinoside (Ara-C) (0.4-1.5 mg/kg every 12 hours for 5 or 7 days). In all these patients, the disease resolved. It is concluded that potentially lethal disease is relatively common in TL, and the available evidence suggests that these diseases are responsive to low-dose Ara-C therapy. PMID- 11902752 TI - Foals of desired sex--and with minimal sperm numbers. PMID- 11902753 TI - Equine clinical cytogenetics--human chromosomes sitting on horse chromosomes. PMID- 11902754 TI - Cultivation and characterisation of primary and subcultured equine keratinocytes. AB - We describe the establishment and characterisation of equine keratinocyte cultures with maintenance of a high proliferative capacity up to the second passage. Improved attachment and growth were obtained by seeding primary cells on equine feeder layers. Subcultured keratinocytes showed optimal growth when seeded on collagen type I. The proliferation rate of cells on this substrate exceeded that seen for cells seeded on equine feeder layers. By immunohistochemistry, epithelial origin and state of differentiation of the equine keratinocytes were determined. They expressed keratin and desmoplakin I/II, but lacked keratin 10. Electron microscopy revealed typical features of cultured keratinocytes. Purity of keratinocyte cultures was determined by vimentin staining. This is the first report on the establishment of equine keratinocytes derived from lip epithelium. It forms the basis to study equine keratinocyte biology and the pathogenesis of epidermal diseases. Since wound healing represents a severe problem in equine dermatology, our data may be essential for the establishment of new and improved therapy. PMID- 11902755 TI - Hysteroscopic insemination of low numbers of flow sorted fresh and frozen/thawed stallion spermatozoa. AB - The objective of this experiment was to determine the effects of flow cytometric sorting and freezing on stallion sperm fertility. A 2 x 2 factorial design was used to delineate effects of flow sorting and freezing spermatozoa. Oestrus was synchronised (July-August) in 41 mares by administering 10 ml altrenogest (2.2 mg/ml) per os for 10 consecutive days, followed by 250 microg cloprostenol i.m. on Day 11. Ovulation was induced by administering 3,000 iu hCG i.v. either 6 h (fresh spermatozoa) or 30 h (frozen/thawed spermatozoa) prior to insemination. Mares were assigned randomly to one of 4 sperm treatment groups. Semen was collected from 2 stallions with an artificial vagina and processed for each treatment. Treatment 1 (n = 10 mare cycles) consisted of fresh, nonsorted spermatozoa and Treatment 2 (n = 16 mare cycles) of fresh, flow sorted spermatozoa. Spermatozoa to be sorted were stained with Hoechst 33342 and sorted into X- and Y-chromosome-bearing populations based on DNA content using an SX MoFlo sperm sorter. Treatment 3 (n = 16 mare cycles) consisted of frozen/thawed nonsorted spermatozoa (frozen at 33.5 x 106 sperm/ml in 0.25 ml straws) and Treatment 4 (n = 15 mare cycles) of flow sorted frozen/thawed spermatozoa (frozen at 64.4 x 10(6) sperm/ml). Concentrations of sperm in both cryopreserved treatments were adjusted, based on predetermined average post-thaw motilities, so that each insemination contained approximately 5 x 10(6) motile spermatozoa. Hysteroscopic insemination of 5 x 10(6) motile spermatozoa in a volume of 230 microd was used for all treatments. Pregnancy was determined ultrasonographically 16 days postovulation. No differences were found (P>0.1) in the pregnancy rates for mares inseminated with fresh nonsorted (4/10 = 40.0%), fresh flow sorted (6/16 = 37.5%), frozen/thawed nonsorted (6/16 = 37.5%) and flow sorted frozen/thawed spermatozoa (2/15 = 133%). Pregnancy rates tended (P = 0.12) to be lower following insemination of frozen/thawed flow sorted spermatozoa. Further studies are needed with a larger number of mares to determine if fertility of flow sorted frozen/thawed spermatozoa can be improved. PMID- 11902756 TI - Water intake and fluid shifts in horses: effects of hydration status during two exercise tests. AB - In the present study, the main objective was to study factors affecting postexercise voluntary water intake in horses. Four Standardbred horses (mean +/- s.e. bwt 500 +/- 8 kg) were used to study water intake and effects of altering hydration status before an incremental exercise test (INCR) and a 40 min constant velocity exercise test (CONST) on a treadmill. Exercise was performed during normohydration (N), after dehydration for 24 h (DEH) and after hyperhydration with 12 l water 30 min before exercise (HH). DEH resulted in a bodyweight loss of 3% and there were signs of some fluid uptake prior to exercise in both HH trials. By the end of the INCR, the calculated change in plasma volume (PVcalc) was -13 +/- 1, -21 +/- 1 and -11 +/- 3% in the N, DEH and HH trials, respectively. During the highest exercise velocities a hypotonic shift of fluid was seen in all INCR trials. There was a greater accumulation of plasma lactate (pLA) in HH-than in N INCR, probably caused by the extra weight to be carried. CONST induced a similar fluid loss (3%) in all trials, but the decrease in PVcalc at the end of exercise was significantly smaller in HH (-7 +/- 2%) than in N (-14 +/- 1%) and DEH (-19 +/- 2%). In DEH-INCR and DEH-CONST, plasma sodium concentration (pNa) was higher than in N until drinking water was offered 1 h postexercise. In the presence of both an increased pNa and a decrease in PVcalc when dehydrated, the horses drank immediately when offered water postexercise. In N-CONST, there was a significant decrease in calculated PVcalc (-10 +/- 2%) but no increase in pNa when water was given and in this trial the horses rehydrated less rapidly. Plasma aldosterone concentration (PAC) had increased to the same magnitude in all trials after about 10 min, irrespective of type of exercise or hydration status. It was concluded that when both an osmotic and hypovolemic thirst stimulus was present, the horses rehydrated more rapidly postexercise. PMID- 11902757 TI - Inhalation of organic dusts and lipopolysaccharide increases gelatinolytic matrix metalloproteinases (MMPs) in the lungs of heaves horses. AB - We report the effects of mouldy hay/straw exposure, inhaled hay dust suspension (HDS) and inhaled lipopolysaccharide (LPS) on bronchoalveolar lavage fluid (BALF) gelatinolytic matrix metalloproteinase (MMP) levels and degree of activation in healthy (n = 6) and heaves- (previously termed chronic obstructive pulmonary disease) affected (n = 6 or 7) horses. Gelatinolytic MMPs in BALF were quantified by zymography, and gelatinases were shown by Western immunoblotting to be MMP-2 and MMP-9. Hay/straw and HDS challenges increased BALF total gelatinolytic activity only in heaves horses, with the majority of gelatinolytic activity comprising pro- and active MMP-9. The 5 h duration hay/straw challenge increased BALF gelatinolytic MMP activity in heaves horses at 5 and 24 h after the start of this challenge, with activity returning to baseline by Day 4. In contrast to hay/straw and HDS challenges, LPS inhalation increased BALF gelatinolytic MMP activity in both groups. For all challenges, absolute BALF neutrophil counts were highly significantly correlated (P<0.0001) with levels of proMMP-9 and active MMP 9, but not with levels of MMP-2 (P>0.05). As gelatinolytic MMPs are pro inflammatory agents, they may contribute to lung dysfunction and tissue destruction in heaves horses exposed to airborne organic stable dusts. PMID- 11902758 TI - The use of magnetic motor evoked potentials in horses with cervical spinal cord disease. AB - The aim of this study was to investigate the use of magnetic motor evoked potentials as an ancillary diagnostic test in horses with cervical cord lesions. Transcranial magnetic stimulation was performed in 12 ataxic horses and the results of the evoked responses were compared to those found in normal horses. The latency and peak-to-peak amplitude of the potentials in the 12 ataxic horses were significantly different from those measured in normal horses. The configuration of the abnormal potentials was also polyphasic. Normalisation of the evoked potentials occurred in none of the horses, presented after a period of clinical improvement. These findings demonstrate that the technique is also able to detect lesions in horses with subtle clinical signs of incoordination. Magnetic transcranial stimulation is a valuable ancillary test to assess the integrity of the motor tracts. The technique is painless and safe and shows good sensitivity to detect lesions along the descending motor pathways. PMID- 11902759 TI - Subjective and quantitative scintigraphic assessment of the equine foot and its relationship with foot pain. AB - It was hypothesised that in solar bone images of the front feet of clinically normal horses, or horses with lameness unrelated to the front feet, there would be less than a 10% difference in the ratio of uptake of radiopharmaceutical in either the region of the navicular bone, or the region of insertion of the deep digital flexor tendon (DDFT), compared to the peripheral regions of the distal phalanx. Nuclear scintigraphic examination of the front feet of 15 Grand Prix show jumping horses, all of which were free from detectable lameness, was performed using dorsal, lateral and solar images. The results were compared with the examinations of 53 horses with primary foot pain, 21 with foot pain accompanying another more severe cause of lameness and 49 with lameness or poor performance unrelated to foot pain. None of the horses with foot pain had radiological changes compatible with navicular disease. All the images were evaluated subjectively. The solar views were assessed quantitatively using regions of interest around the navicular bone, the region of insertion of the deep digital flexor tendon and the toe, medial and lateral aspects of the distal phalanx. In 97% of the feet of normal showjumpers, there was <10% variance of uptake of the radiopharmaceutical in the navicular bone, the region of insertion of the DDFT and the peripheral regions of the distal phalanx. There was a significant difference in uptake of radiopharmaceutical in the region of the navicular bone in horses with foot pain compared to normal horses. There was a large incidence of false positive results related to the region of insertion of the DDFT. Lateral pool phase images appeared more sensitive in identifying potentially important DDFT lesions. There was a good correlation between a positive response to intra-articular analgesia of the distal interphalangeal joint and intrathecal analgesia of the navicular bursa and increased uptake of radiopharmaceutical in the region of the navicular bone in the horses with primary foot pain. It is concluded that quantitative scintigraphic assessment of bone phase images of the foot, in combination with local analgesic techniques, can be helpful in the identification of the potential source of pain causing lameness related to the foot, but false positive results can occur, especially in horses with low heel conformation. PMID- 11902760 TI - Changes in fibre type composition of gluteus medius and semitendinosus muscles of Dutch Warmblood foals and the effect of exercise during the first year postpartum. AB - In order to obtain broader insights into the equine musculoskeletal system, we studied the fibre type composition of 2 locomotory muscles in biopsies from Dutch Warmblood foals taken at 3 different ages in the first year postpartum. The muscle fibre types were determined histochemically as well as immunohistochemically. ATPase-characterised IIB fibres appear to express either IId or type lIa plus IId myosin heavy chain (MHC). A high percentage of fibres classified as IIA with ATPase expressed both fast types of MHC. The type I classification by the 2 methods matched almost completely. There was an increase with age of fibres expressing I and IIa MHC in the gluteus medius. At the same time, there was a decrease of fibres expressing IId MHC and fibres co-expressing MHC IIa and IId. MHC expression of the semitendinosus muscle did not change over time at first, but from age 22-48 weeks there was a decrease in the percentage of type IId fibres. In general, the gluteus medius contained more type I fibres but fewer type IId fibres compared to the semitendinosus. At most ages the fibre type compositions of both muscles correlated with one another. To examine the effect of exercise, one-third of the foals were given box rest, one-third received training and one-third kept at pasture during the first 22 weeks of life. The 3 exercise groups differed in their fibre type composition; however, these differences could not be attributed to the effect of exercise. PMID- 11902761 TI - A comparison of three horseshoeing styles on the kinetics of breakover in sound horses. AB - A variety of horseshoe designs are believed to 'ease' breakover, or the unloading of the foot once the heels leave the ground. In this study, conventional toe-clip shoes, quarter-clip shoes, fitted to the white line at the toe, and Natural Balance horseshoes were fitted to the front feet of 9 sound Irish Draught-cross type horses. Forceplate and video motion analyses were undertaken during trot locomotion to determine the moment arm of the ground reaction force on the distal interphalangeal (DIP) joint, the peak DIP joint moment and the peak compressive force on the navicular bone. DIP joint moment arm during breakover was reduced with both Natural Balance (mean +/- s.d. 77 +/- 7 mm) and quarter-clip shoes (78 +/- 9 mm) compared to the toe-clip shoes (86 +/- 6 mm) (P<0.01). Peak DIP joint moment was not significantly different (175 +/- 37,171 +/- 38 and 175 +/- 31 Nmm/kg, in Natural Balance, quarter-clip and toe-clip shoes, respectively) and neither was peak force on the navicular bone (5.52 +/- 1.52, 5.79 +/- 1.53 and 6.14 +/- 1.47 N/kg, respectively). Breakover duration (heel off to toe off) was not significantly reduced by the Natural Balance shoes (39 +/- 6 ms) or the quarter-clip shoes (40 +/- 6 ms) compared to toe-clip shoes (42 +/- 9 ms). This study has demonstrated that the use of Natural Balance shoes reduces the moment arm of the ground reaction force (GRF) during breakover but does not reduce the peak DIP joint moment or the force on the navicular bone. PMID- 11902762 TI - The effect of a pectin-lecithin complex on prevention of gastric mucosal lesions induced by feed deprivation in ponies. AB - This study examined whether a product containing a pectinlecithin complex (Pronutrin) (1) could prevent gastric lesions induced in the equine gastric squamous epithelial mucosa using a protocol of intermittent feed deprivation that resulted in prolonged increased gastric acidity (Murray and Eichorn 1996). Eight ponies were used and served as their own controls in 2 trials in which there were 72 h cumulative deprivation (alternating 24 h with no feed, then 24 h free choice hay), with a 4-week interval between trials. Ponies were assigned randomly to receive either 250 g Pronutrin plus 200 g pelleted feed, or 450 g pelleted feed only. Ponies were conditioned to each treatment for 7 days and received Pronutrin and pellets or only pellets once daily during the feed deprivation protocol. Gastroscopy was performed at the beginning and conclusion of the feed deprivation protocol. The endoscopist (MJ.M.) was blinded as to treatments, and lesion severity was scored on a scale of 0-5. Gastroscopy revealed normal-appearing gastric mucosa at the beginning of feed deprivation, with the exception of 2 ponies which had focal squamous mucosal erosion and 1 pony with focal glandular mucosal erosion. After 72 h cumulative feed deprivation, each pony, except 1 pony in one of the trials, developed erosions or ulcers in the gastric squamous mucosa. There was no difference (P = 0.6) in the presence or severity of gastric lesions between treatments. Lesions did not develop in the gastric glandular mucosa as a result of the intermittent feed deprivation with either treatment. In this study, the pectin-lecithin complex in Pronutrin failed to prevent lesions in the gastric squamous mucosa induced by intermittent feed deprivation. PMID- 11902763 TI - Anatomical study of the notches in the nasal process of the equine incisive bone. PMID- 11902765 TI - Use of zoo-FISH to characterise a reciprocal translocation in a thoroughbred mare: t(1;1 6)(q16;q21.3). PMID- 11902764 TI - Detection of high circulating concentrations of inhibin pro- and -alphaC immunoreactivity in mares with granulosa-theca cell tumours. PMID- 11902766 TI - Osteochondrosis of the second cervical vertebra of a horse. PMID- 11902768 TI - A neural network model to predict the wastewater inflow incorporating rainfall events. AB - Under steady-state conditions, a wastewater treatment plant usually has a satisfactory performance because these conditions are similar to design conditions. However, load variations constitute a large portion of the operating life of a treatment facility and most of the observed problems in complying with permit requirements occur during these load transients. During storm events upsets to the different physical and biological processes may take place in a wastewater treatment plant, and therefore, the ability to predict the hydraulic load to a treatment facility during such events is very beneficial for the optimization of the treatment process. Most of the hydrologic and hydraulic models describing sewage collection systems are deterministic. Such models require detailed knowledge of the system and usually rely on a large number of parameters, some of which are uncertain or difficult to determine. Presented in this paper, an artificial neural network (ANN) model that is used to make short term predictions of wastewater inflow rate that enters the Gold Bar Wastewater Treatment Plant (GBWWTP), the largest plant in the Edmonton area (Alberta, Canada). The neural model uses rainfall data, observed in the collection system discharging to the plant, as inputs. The building process of the model was conducted in a systematic way that allowed the identification of a parsimonious model that is able to learn (and not memorize) from past data and generalize very well to unseen data that was used to validate the model. The neural network model gave excellent results. The potential of using the model as part of a real-time process control system is also discussed. PMID- 11902767 TI - Removal of phenol from water by adsorption-flocculation using organobentonite. AB - Bentonite modified with short chain cationic surfactant might be the basis of a new approach to removing organic pollutants from water. The treatment process involves dispersing bentonite to the contaminated water and then adding a small cationic surfactant so as to result in flocs which are agglomerates of organobentonite and bound organic pollutants. The flocs are then removed from the solution by sedimentation. Experimental results indicate that BTMA-bentonite displays a high affinity for phenol, possibly because phenol molecules interact favorably with the benzene ring in BTMA ion through increased pi-pi type interactions. Under appropriate operating conditions, 90% phenol removal and nearly 100% bentonite recovery could be achieved by the adsorption flocculation process using BTMA-bentonite. Additionally, the insensitivity of the process to the changing ionic strength of the solution and rapid adsorption kinetics made adsorption-flocculation with BTMA-bentonite attractive for continuous treatment of large volumes of industrial wastewater. The bentonite may function as a recyclable surfactant support for the adsorption and subsequent combustion of organic pollutants. PMID- 11902769 TI - Complete physico-chemical treatment for coke plant effluents. AB - Naturally found coal is converted to coke which is suitable for metallurgical industries. Large quantities of liquid effluents produced contain a large amount of suspended solids, high COD, BOD, phenols, ammonia and other toxic substances which are causing serious pollution problem in the receiving water to which they are discharged. There are a large number of coke plants in the vicinity of Jharia Coal Field (JCF). Characteristics of the effluents have been evaluated. The present effluent treatment systems were found to be inadequate. Physico-chemical treatment has been considered as a suitable option for the treatment of coke plant effluents. Ammonia removal by synthetic zeolite, activated carbon for the removal of bacteria, viruses, refractory organics, etc. were utilized and the results are discussed. A scheme has been proposed for the complete physico chemical treatment, which can be suitably adopted for the recycling, reuse and safe disposal of the treated effluent. Various unit process and unit operations involved in the treatment system have been discussed. The process may be useful on industrial scale at various sites. PMID- 11902770 TI - P-nitrophenol degradation by activated sludge attached on nonwovens. AB - p-Nitrophenol (PNP) is a toxic compound that enters the environment during manufacturing and processing of a variety of industrial products. This study demonstrates the use of inexpensive and durable nonwovens as a biomass retainer for the biological degradation of p-nitrophenol. An essential aspect of p nitrophenol degradation was the cultivation of p-nitrophenol degrading biomass prior to its attachment on the nonwovens. Results of continuous flow experiments demonstrated that using the nonwovens could attain consistent high-rate p nitrophenol degradation. 500 mg-PNP/L was completely degraded at a hydraulic retention time of 11 h. Specific and volumetric p-nitrophenol loading rates were determined to be 165 mg-PNP/g-MLSS/d and 1.6 g-PNP/L/d, respectively. Nitrite released from p-nitrophenol breakdown was not completely nitrified to nitrate. Characteristics of p-nitrophenol degrading sludge were also investigated. PMID- 11902771 TI - Advanced oxidation of a reactive dyebath effluent: comparison of O3, H2O2/UV-C and TiO2/UV-A processes. AB - In the present study the treatment efficiency of different AOPs (O3/OH- H2O2/UV-C and TiO2/UV-A) were compared for the oxidation of simulated reactive dyebath effluent containing a mixture of monochlorotriazine type reactive dyes and various dye auxiliary chemicals at typical concentrations encountered in exhausted reactive dyebath liquors. A525 (color), UV280 (aromaticity) and TOC removal rates were assessed to screen the most appropriate oxidative process in terms of reactive dyebath effluent treatment. Special emphasis was laid on the effect of reaction pH and applied oxidant (O3, H2O2) dose on the observed reaction kinetics. It was established that the investigated AOPs were negatively affected by the Na2CO3 content (= 867 mg/L) which is always present at high concentrations in dychouse effluents since it is applied as a pH buffer and dye fixation agent during the reactive dyeing process. The ozonation reaction exhibited almost instantaneous decolorization kinetics and a reasonable TOC reduction rate. It appeared to be stable under the investigated advanced oxidation conditions and outranked the other studied AOPs based on the above mentioned criteria. Besides, the electrical energy requirements based on the EE/O parameter (the electrical energy required per order of pollutant removal in 1 m3 wastewater) was calculated for the homogenous AOPs in terms of decolorization kinetics. In view of the electrical energy efficiency, ozonation and H2O2/UV-C oxidation at the selected treatment conditions appear to be promising candidates for full-scale dyehouse effluent decolorization. PMID- 11902772 TI - Drinking water denitrification by a membrane bio-reactor. AB - Drinking water denitrification performance of a bench scale membrane bio-reactor (MBR) was investigated as function of hydraulic and biological parameters. The reactor was a stirred tank and operated both in batch and continuous mode. The mixed denitrifying culture used in the batch mode tests was derived from the mixed liquor of a wastewater treatment plant in Erzincan province in Turkey. But the culture used in the continuous mode tests was that obtained from the batch mode tests at the end of the denitrification process. The nitrate contaminated water treated was separated from the mixed liquor suspended solids (MLSS) containing active mixed denitrifying culture and other organic substances by a membrane of 0.2 microm average pore diameter. The results indicated that the use of a membrane module eliminated the need for additional post treatment processes for the removal of MLSS from the product water. Concentration of nitrite and that of MLSS in the membrane effluent was below the detectable limits. Optimum carbon to nitrogen (C/N) ratio was found to be 2.2 in batch mode tests. Depending on the process conditions, it was possible to obtain denitrification capacities based on the reactor effluent and membrane effluent up to 0.18kgm(-3)day(-1) and 2.44 kg m(-2) day2(-1) NO(3-)-N, respectively. The variation of the removal capacity with reactor dilution rate and membrane permeate flux was the same for two different degrees of [MLSS]0/[NO3-N]0 (mass) ratios of 25.15 and 49.33. The present MBR was able to produce a drinking water with NO(3-)-N concentration of less than 4 ppm from a water with NO3-N contamination level of 367 ppm equivalent to a NO(3-)-N load of 0.310 kgm(-3) day(-1). The results showed that MBR system used was able to offer NO(3-)-N removals of up to 98.5%. It was found that the membrane limiting permeate flux increased with increasing MLSS concentration. PMID- 11902773 TI - Accuracy analysis of a respirometer for activated sludge dynamic modelling. AB - The aim of the paper is to assess the experimental errors arising from the operation of a closed respirometer using autotrophic biomass. A closed, intermittent-flow device has been set-up for the measurement of oxygen uptake rate (OUR) and parameter calibration. After describing the device structure and operation, the factors affecting accuracy have been assessed. Inaccuracies may be caused by two groups of parameters: design parameters, including flow rate, volume, sampling time, numerical algorithm, sample injection and environmental parameters, concerning the physicochemical conditions of the experiment, such as unwanted oxygen transfer, pH, and the influence of sludge condition on "start-up" behaviour. It is shown to what extent each of them affects the final accuracy of the OUR measurement. In the second part of the paper, the respirometric data are used to calibrate a two-step nitrification model and their impact on the accuracy of the estimation of model parameters is assessed. Confidence limits are derived for the identifiable parameter combinations and the practical identifiability assessed with the aid of trajectory sensitivity analysis. PMID- 11902774 TI - Factors affecting seasonal variation of membrane filtration resistance caused by Chlorella algae. AB - A seasonal fluctuation pattern was observed in membrane filtration resistance by Chlorella algae cultured in open ponds in the tropical environment. In order to investigate the causes of this phenomenon, Chlorella was cultivated under controlled conditions and the cake resistance was measured by batch filtration in dead-end mode. The filtration resistance was found to be a function of environmental conditions. Algae could grow favourably and offered low specific cake resistance (R,s) on the order of 10(11) m/g for the culture temperature from 28 degrees C to 35 degrees C. The algal growth was inhibited and the specific cake resistance increased to the order of 10(12) m/g below or above this optimum temperature range. Strong solar radiation, coupled with high temperatures, also inhibited the growth of algae and resulted in higher specific cake resistance. The specific cake resistance of algae cultured at different temperatures increased with the amount of the extracellular organic matter (EOM) extracted by 0.1 N NaOH. Hence EOM, rather than bacteria present in the mono-algal culture, was considered to be the primary factor affecting the cake resistance. The specific cake resistance increased drastically after actively growing cells were stored in nutrient-free water under dark conditions. However, the resistance was slightly decreased when the algal cells were stored in NSIII nutrient media in a dark room, indicating the effect of nutrient availability on the change of the specific cake resistance under the light-limiting conditions. EOM extracted from the cells kept in the nutrient-free water contained less sugar than the fresh culture, whereas the EOM extracted from the cells stored in the NSIII media contained more sugar. The molecular distribution of the EOM shifted from below 1,000 kDa before storage to more than 2,000 kDa after storage in both the nutrient-free and NSIII media. PMID- 11902775 TI - The efficiency of oxidation ponds at the Kraft pulp and paper mill at Webuye in Kenya. AB - Two streams of wastewater from the Kraft Pulp and Paper Mills at Webuye are released from the pulping, bleaching and paper pressing departments. One stream is made up of clear wastewater and the second is made up of turbid wastewater. The wastewaters from the mills are treated using oxidation ponds, after which the treated wastewater is discharged into the River Nzoia. The wastewater was analysed for Cd, Cr, Cu, Ni, Pb. Zn, Ca, K, Mg and Na. The results are that the concentrations of Ca and Na significantly increased in the treated wastewater. The concentrations of Cd, Pb, Zn, K and Mg also increased in the treated wastewater, though not significantly. The concentrations of Cr, Cu and Ni decreased in the treated wastewater but not significantly. Mass loadings of the metals discharged into River Nzoia were calculated. It was found that the mean mass loading of two metals (Ca and Na) significantly increased in the treated wastewater. Five metals (Cd, Pb, Zn, K and Mg) increased in the treated wastewater although not significantly. Three metals (Cr, Cu and Ni) decreased in the treated wastewater but the decrease was not significant. The concentrations of Cd, Cu, Ni, Pb and Zn were all above the limits set by EC for drinking water. These findings pose serious questions of the effectiveness of oxidation ponds system to safeguard the receptor against pollutants from the pulp and paper industry at Webuye. PMID- 11902776 TI - Migration of dissolved heavy metal compounds and PCP in the presence of colloids through a heterogeneous calcareous gravel and a homogeneous quartz sand-pilot scale experiments. AB - In preparation to field experiments with in situ mobilized colloids, a set of pilot scale aquifer tank experiments was performed to gain an insight into the mass transfer of several heavy metal ions and pentachlorophenol (PCP) in a heterogeneous gravel aquifer and a homogeneous sandy aquifer. The experiments simulate a spill of dissolved contaminants and their subsequent transport with in situ mobilized colloids that are not yet in equilibrium with the contaminant. Contaminants with a weak tendency to adsorb to the stationary sediment matrix, e.g., PCP, establish a dynamic equilibrium with the mobile colloidal phase. The fraction of contaminants sorbed to colloids remains constant along the observed flow path. In contrast, for contaminants that strongly sorb to the sediment matrix, the fraction associated with colloids increases with increasing flow distance. In either case, the filtration of colloids is the limiting factor for the colloidal mass transfer. It is also shown that the recovery rates decrease rapidly within both sediments, and that the transport parameters (flow velocity, dispersivity) are only representative for the mobile fraction of the contaminant. On the given experimental scale, there was no evidence of a preferred colloidal transport, or even a transport of colloids ahead of the conservative tracer. PMID- 11902777 TI - Studies on enhancement of Cr(VI) biosorption by chemically modified biomass of Rhizopus nigricans. AB - This study reports the biosorption of Cr(VI) by chemically modified biomass of Rhizopus nigricans and the possible mechanism of Cr complexation to the adsorbent. The cell wall of this fungus possesses strong complexing property to effectively remove Cr(VI) anions from solution and wastewater. The mechanism of Cr adsorption by R. nigricans was ascertained by chemical modifications of the dead biomass followed by FTIR spectroscopic analysis of the cell wall constituents. Treatment of the biosorbent with mild alkalies (0.01 N NaOH and ammonia solution) and formaldehyde (10%, w/v) deteriorated the biosorption efficiency. However, extraction of the biomass powder in acids (0.1 N HCl and H2SO4), alcohols (50% v/v, CH3OH and C2H5OH) and acetone (50%, v/v) improved the Cr uptake capacity. Reaction of the cell wall amino groups with acetic anhydride reduced the biosorption potential drastically. Blocking of the-COOH groups by treatment with water soluble carbodiimide also resulted in initial lag in Cr binding. Biomass modification experiments conducted using Cetyl Trimethyl Ammonium Bromide (CTAB), Polyethylenimine (PEI), and Amino Propyl Trimethoxy Silane (APTS) improved the biosorption efficiency to exceptionally high levels. The FTIR spectroscopic analysis of the native, Cr bound and the other types of chemically modified biomass indicated the involvement of amino groups of Rhizopus cell wall in Cr binding. The adsorption data of the native and the most effectively modified biomass were evaluated by the Freundlich and the Langmuir adsorption isotherms and the possible adsorption phenomena are also discussed. PMID- 11902778 TI - Ozonation of naphthalene sulfonic acids in aqueous solutions. Part I: elimination of COD, TOC and increase of their biodegradability. AB - Ozonation of 11 naphthalene sulfonic acids (NSA) in the aqueous solution was studied by bubbling at 31 degrees C at an ozone dose rate of 5.56 mg min(-1) l( 1). COD, TOC and BOD5 of these compounds were tested. It was found that COD and TOC can be removed effectively by ozonation as expected. More than 40% COD of compounds No. I (2-amino-1-NSA), No. 5 (1-hydroxy-7-amino-5-NSA), No. 6 (6 hydroxy-1-NSA), No. 8 (6-amino-1,4-naphthalene disulfonic acid) and No. 11 (I hydroxy-6-amino-3-NSA) has been eliminated at an ozone dosage of 5.56 mg min(-1) l(-1) for 2 h. Although TOC removal was very different, a good biodegradability was reached for NSAs with an average ozone consumption of 3.0 mgl(-1) for a TOC0 concentration of 100 mg(-1). Ozonation and biotreatment should be good alternatives for these compounds, especially after 20% TOC reduction and TOC removal were more sensitive in predicting an increase in biodegradability during ozonation, than that of COD. NSAs are similar in their behavior with reference to ozone consumption. In order to obtain a good biodegradability of NSAs at a TOC0 concentration of 100 mgl(-1), an ozone consumption between 2.0 and 4.0 mg mg(-1) ACOD is needed for this setup and 3.0 mg O3 mg(-1) TOC0 requirement may be more practical in predicting the biological behavior of naphthalene compounds. PMID- 11902779 TI - Adsorption of cadmium by activated carbon cloth: influence of surface oxidation and solution pH. AB - The surface of activated carbon cloth (ACC), based on polyacrylonitrile fibre as a precursor, was oxidised using nitric acid, ozone and electrochemical oxidation to enhance cadmium ion exchange capacity. Modified adsorbents were physically and chemically characterised by pH titration, direct titration, X-ray photoelectron spectroscopy, elemental analysis, surface area and porosimetry, and scanning electron microscopy. BET surface area decreased after oxidation, however, the total ion exchange capacity increased by a factor of approximately 3.5 compared to the commercial as-received ACC. A very significant increase in cadmium uptake, by a factor of 13, was observed for the electrochemically oxidised ACC. Equilibrium sorption isotherms were determined at pH 4, 5 and 6 and these showed that cadmium uptake increased with increasing pH. There was clear evidence of physical damage to ozone-oxidised fibre, however, acid and electrochemically oxidised samples were completely stable. PMID- 11902780 TI - Nonionic surfactant effects on pentachlorophenol biodegradation. AB - Several potential mechanisms of surfactant-induced inhibition of pentachlorophenol (PCP) biodegradation were tested using a pure bacterial culture of Sphingomonas chlorophenolicum sp. Strain RA2. PCP degradation, glucose degradation, and oxygen uptake during endogenous conditions and during glucose degradation were measured for batch systems in the presence of the nonionic surfactant Tergitol NP-10 (TNP10). TNP10 did not exert toxicity on RA2 as measured by dissolved oxygen uptake rates under endogenous conditions and glucose biodegradation rates. TNPIO reduced the substrate inhibition effect of PCP at high PCP concentrations, resulting in faster PCP degradation rates at higher concentrations of TNP10. Calculations of a micelle partition coefficient (Kmic) show that PCP degradation rates in the presence of surfactant can be explained by accounting for the amount of PCP available to the cell in the aqueous solution. A model is discussed based on these results where PCP is sequestered into micelles at high TNP10 concentrations to become less available to the bacterial cell and resulting in observed inhibition. Under substrate toxicity conditions, the same mechanism serves to increase the rate of PCP biodegradation by reducing aqueous PCP concentrations to less toxic levels. PMID- 11902781 TI - A hydro-chemical study of a mountainous watershed: the Ganga, India. AB - A hydro-chemical study has been carried out on a 37-km stretch of the River Ganga from Deoprayag to Rishikesh (India) during the period from April 1999 to March 2000. The assessment of sediment and nutrient load has been considered to evaluate the current state of pollution through real time measurements. The values of pH and conductance are well within the limits prescribed for drinking water. The maximum suspended sediment concentrations of 1,405 and 2,002 mg/L were recorded at Deoprayag and Rishikesh, respectively, during the rainy season. A large amount of sediment and nutrient load is transported from the watershed during the rainy season. Concentrations of N(O3-)-N and N(H3-)-N at Deoprayag varied from 0.30 to 0.50 and 0.02 to 0.12 mg/L, respectively, depending on season. Examination of the results showed clearly that N(H3-)-N was generally low as compared to N03-N. Depending on the pH and temperature of soils, NH4+ and NO3- ions are produced in the watershed through ammonification and nitrification of organic matter and mobilized into rivers through run-off. Dissolved N and P from fertilizer application, sewage and non-point source run-off contribute significant quantities of these nutrients in river water. The nitrate and phosphate are transported from the cropland either by being adsorbed on to soil particles that are subsequently eroded, or dissolved in runoff water from agricultural land. The data generated through the study will be useful for development and management planning of the hilly watershed. PMID- 11902783 TI - Methods for preparing synthetic freshwaters. AB - Synthetic solutions that emulate the major ion compositions of natural waters are useful in experiments aimed at understanding biogeochemical processes. Standard recipes exist for preparing synthetic analogues of seawater, with its relatively constant composition, but, due to the diversity of freshwaters, a range of compositions and recipes is required. Generic protocols are developed for preparing synthetic freshwaters of any desired composition. The major problems encountered in preparing hard and soft waters include dissolving sparingly soluble calcium carbonate, ensuring that the ionic components of each concentrated stock solution cannot form an insoluble salt and dealing with the supersaturation of calcium carbonate in many hard waters. For acidic waters the poor solubility of aluminium salts requires attention. These problems are overcome by preparing concentrated stock solutions according to carefully designed reaction paths that were tested using a combination of experiment and equilibrium modeling. These stock solutions must then be added in a prescribed order to prepare a final solution that is brought into equilibrium with the atmosphere. The example calculations for preparing hard, soft and acidic freshwater surrogates with major ion compositions the same as published analyses, are presented in a generalized fashion that should allow preparation of any synthetic freshwater according to its known analysis. PMID- 11902782 TI - Determination of aliphatic hydrocarbons in urban runoff samples from the "Le Marais" experimental catchment in Paris centre. AB - Aliphatic hydrocarbons were assessed in runoff and waste waters from an urban catchment located in Paris. Runoff were sampled from different types of urban surfaces (11 roofs representing four different covering materials, two courtyards and six streets). Waste water samples were collected at the catchment outlet during dry and wet weather periods as well. This paper gives an overview of the results on the concentration and distribution points of view for both the particulate and the dissolved phases. Results were discussed on the basis of the median. Accordingly, the temporal variability was taken into account. Thus, the concentration medians ranged from 345 to 827, from 297 to 790, and from 393 to 1,359 microg L(-1) in the roof, courtyard and street samples, respectively. The levels found at the catchment outlet during dry and wet weather periods were of the same order of magnitude, i.e. 700 microgL(-1). The particulate phase represented 85% of the total aliphatic hydrocarbon content whatever the sample. PMID- 11902785 TI - Removal of phosphate by layered double hydroxides containing iron. AB - Iron-based layered double hydroxides (M2+(a)Fe3+(b) (OH)2(a+b) CO3(2-) b/2mH2O) were synthesized. Removal of phosphate by the compounds was studied from the viewpoint of buffering pH effect of the compounds and buffering capacity of solution. The compounds released metal cations (Mg2+, Ca2 , Fe3+) and/or their hydroxides responding to various water environments due to their buffering pH function. The released cations and/or hydroxides worked effectively as coagulants for the phosphate removal. The removal of phosphate depended on the buffering capacity of the solution that is the function of the solution pH and the concentration of phosphate. The removal of phosphate from the solution with small buffering capacity followed a Langmuir-type isotherm. The removal of phosphate from the solution with larger buffering capacity was largely increased. The removal of phosphate by the compounds was analyzed based on the model describing the buffering pH effect of the compounds from which the amount of released cations (CB) can be determined. The removal was well correlated with the amount of dissolution of the compounds, CB. The mechanism of phosphate removal was examined based on the removal efficiency (mol of removed phosphate/mol of released alkali). The efficiencies showed below one in the solution with large buffering capacity and above one in the solution with small buffering capacity. The efficiency below one showed the removal of phosphate through coagulation by the released metal cations and hydroxides. The successful removal of dilute phosphate (0.2mg P/l) from the drain water was also demonstrated. PMID- 11902784 TI - Calorimetry: a tool for assessing microbial activity under aerobic and anoxic conditions. AB - For many years, calorimetric measurements have been used for understanding, modelling, controlling, and optimising chemical reactions. Calorimetry could be as well utilised to investigate biological processes, which however, involve very small amount of heat and therefore require very sensitive instruments. For this purpose, a Mettler Toledo RCI (Reaction calorimeter) was modified, changing both hardware and software, increasing its resolution up to 5 10m W/l. Such sensitivity allows the monitoring of aerobic and anoxic processes. This paper points out the excellent agreement between calorimetric and respirometric data, obtained simultaneously under aerobic conditions using activated sludge from a lab-scale scale reactor. Heat production rate can be directly converted in oxygen uptake rate by means of a correlation factor, whose value is approximately the same for all aerobic respiratory metabolisms. Taking into account this factor, calorimetric data were introduced in a chemical oxygen demand based model and processed for the estimation of kinetic parameters of heterotrophic biomass. Aerobic heterotrophic, denitrifying, and autotrophic nitrifying activity were determined by specific calorimetric tests. The effect of potentially toxic or inhibitory substances on the activity of all microbial communities was as well pointed out in these measurements. PMID- 11902786 TI - Derivation and application of a new model for heavy metal biosorption by algae. AB - An equilibrium model for describing the relationships between important parameters for heavy metal sorption by algae was derived through a thermodynamics approach. In this model, both the removal efficiency of heavy metal and metal adsorption per unit algal biomass are considered to be simple functions of the ratio of algal biomass concentration to the initial metal concentration for selected conditions, i.e. as at constant pH and temperature. The model was found to fit the experimental results well (judged by the correlation-regression coefficient, R2), for the adsorption of cadmium, copper, lead and zinc by two algal species, Oocystis sp. (both living and non-living) and Chlorococcum sp. The applicability of the model was also supported by the reprocessed results of experimental data given in the literature, i.e. for the metal species, Cd, Pb, Cu and Ag, the algal species, Chlorella vulgaris, Scenedesmus quadricauda and Cladophora crispata, and both batch and continuous fixed-bed reactors. It was also demonstrated that the model could be applied over a broad range of pH for cadmium and copper adsorption by Oocystis sp. However, the model was not applicable at very low and high pH levels, due to negligible adsorption and precipitation, respectively. PMID- 11902787 TI - Phosphorus removal using cow bone in hydroxyapatite crystallization. AB - The applicability of cow bone as a seed material in hydroxyapatite crystallization was investigated in this research. Seed crystal used for the experiments was prepared by the calcination of cow bone. The effects of initial calcium concentration, pH, alkalinity, reaction temperature condition, and calcination temperature were examined for synthetic solution by batch experiment. The experimental results showed that a good phosphorus removal could be achieved by cow bone crystal seeding. The phosphorus removal rate with various calcium concentrations and pHs could be predicted from high relationships between residual phosphorus concentration and pH (after reaction). The effects of alkalinity, reaction temperature condition, and calcination temperature were also examined by rate constant analysis. PMID- 11902788 TI - Fenton oxidation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and octahydro 1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX). AB - Oxidation of the high explosives hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1.3,5,7-tetrazocine (HMX) using Fenton's reagent proceeds rapidly between 20 degrees C and 50 degrees C at pH 3. At an H2O2: Fe2+: RDX molar ratio of 5,178: 48: 1, RDX and HMX were completely removed in 1 to 2 h. All the experimental data could be fit to a pseudo first-order rate equation. The reaction rate was also strongly dependent on Fenton's reagent concentrations. NO3 and N2 were identified as nitrogen byproducts from RDX and HMX oxidation. The experiment with radiolabeled RDX showed that approximately 37% of organic carbon in RDX was mineralized to CO2. We observed formaldehyde and formic acid as a short-lived intermediate. No other volatile or nonvolatile byproducts were found from GC/MS analysis. The results show that RDX and HMX can be effectively mineralized with Fenton's reagents. PMID- 11902789 TI - Ultrasound pretreatment of elemental iron: kinetic studies of dehalogenation reaction enhancement and surface effects. AB - This work presents data showing the kinetic improvement afforded by ultrasound pretreatment and illustrates the physical and chemical changes that take place at the iron surface. First-order rate constants improved as much as 78% with 2h of ultrasound pretreatment. Scanning electron microscopy (SEM) and surface area analysis were used for confirmation of the physical changes that take place after ultrasound was used on iron surfaces exposed to a variety of conditions. X-ray photoelectron spectroscopy was used to determine chemical surface characteristics before and after ultrasound use. SEM and surface area analysis showed that ultrasound use clears the iron surface of debris increasing the surface area up to 169%. In addition, exposure to ultrasound alters ratios of surface species, such as adventitious carbon to carbonyl carbon and iron to oxygen, and removed hydroxides thus making the iron more reactive to reductive dehalogenation. PMID- 11902790 TI - Identification and modelling of dry weather processes in gully pots. AB - This paper presents the results of a monitoring and modelling programme, carried out to study the processes occurring in gully pots during dry weather. The monitoring programme involved estimation of the change in gully pot liquor quality, under field and laboratory conditions. The contents (i.e. liquor and sludge) of pots draining five different types of roads were monitored over dry periods in the winter and summer. A bench scale study was carried out to study the influence of temperature variations and sludge digestion by-products on gully liquor quality. The change in quality was measured in terms of chemical oxygen demand (COD), dissolved oxygen (DO) and ammonium concentration. The trends for change in COD and DO, were found to be broadly similar for all road types. However, ammonium transformation was found to follow different patterns at different locations. Several dry weather processes, such as COD decay, ammonium transformation, oxygen depletion and uptake by sludge, oxygen transfer from the atmosphere and benthic release of COD, have been identified. Other processes, found to take place during dry weather, include development of a scum layer over the surface of the gully liquor and sludge bulking. A model is proposed which can predict the change in gully liquor quality in terms of COD, DO and ammonium concentrations. Data collected during the study has been used to successfully calibrate and verify the model. PMID- 11902791 TI - Modelling of the retention of uncharged molecules with nanofiltration. AB - In this paper, a model is developed for the retention of organic molecules with a given nanofiltration membrane at different pressures as a function of the molecular weight. The Spiegler-Kedem transport equations were used to derive the reflection coefficient, the maximal retention that would theoretically be obtained at infinite transmembrane pressure, from experimental retention values for a large set of molecules with the effective diameter of the molecule as a size parameter. Secondly, the pore size distribution of the membrane is derived from the experimental reflection coefficients. This allows to calculate the reflection coefficient for a molecule with a given effective diameter. Since this parameter is not readily available, a correlation between the effective diameter and the molecular weight has been established and introduced in the model equations. Subsequently, the contribution of diffusion in the transport of molecules through the membrane was evaluated by introducing a membrane diffusion parameter, which was determined experimentally for the membranes NF70, NTR 7450 and UTC-20. Finally, the pore size distribution, the diffusion parameter and the experimental water flux through the membrane were used to calculate the retention as a function of the molecular weight and pressure for the same three membranes. This allows to determine retention curves at different pressures, and to calculate the variation of the MWC with pressure. PMID- 11902792 TI - Degradation of 2,4-dichlorophenoxyacetic acid by ionizing radiation: influence of oxygen concentration. AB - Ionizing radiation has been proved as a promising method for the degradation of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The gamma-radiolytic decomposition and chloride formation follows an apparent first order kinetic, kde/kCl = 3. For complete removal of 500 microM 2,4-D, a dose of 4 kGy is required. Phenolic intermediates (2,4-dichlorophenol, the isomers 2-chloro-4 hydroxy- and 4-chloro-2-hydroxy phenoxyacetic acid and three hydroxylation products of 2,4-D) are formed. Using oxygen saturation during irradiation, they are removed again with a dose of 4 kGy. For the formation of the main organic product acetic acid a reaction mechanism is discussed. Oxygen concentration enhances strongly fragmentation and mineralization. A reduction of 36% TOC could be achieved. PMID- 11902793 TI - A molecular typing approach for evaluating bioaerosol exposure in wastewater treatment plant workers. AB - Bioaerosols associated with wastewater treatment process may represent a health risk for occupationally exposed personnel. To evaluate microbial contamination in plant workers, we compared oral cavity isolates against isolates collected from aerosol surrounding the aeration basin. Typing was performed by metabolic profile and arbitrarily primed-polymerase chain reaction. The latter is more sensitive and rapid than conventional tests. After comparison, isolates from the air samples were not related to those obtained from the exposed workers. This molecular approach can support bioaerosol risk evaluation . PMID- 11902794 TI - Removal of THM precursors by GAC: Ankara case study. AB - The effectiveness of granular activated carbon (GAC) adsorption for the removal of natural organic matter and trihalomethanes from Ivedik Water Treatment Plant of Ankara City is investigated. Freundlich Isotherm constants K and n were determined as 17.61 and 1.66, respectively. Bench-scale GAC columns were run with empty bed contact times (EBCT) varying from 0.40 to 2.67 min to evaluate adsorption performance. 50% exhaustion values were used for comparison. The treated volumes of water increased with EBCT, showing a linear increase in GAC service life. Correspondingly, the carbon usage rate decreased. The capacities calculated by the isotherm equation and achieved by columns were also compared. The column capacities were within 43-65% of the isotherm capacities at complete breakthrough. However, they were only within 8-17% of the isotherm capacities at 50% breakthrough. PMID- 11902795 TI - Methane yield as a monitoring parameter for the start-up of anaerobic fixed film reactors. AB - This paper describes the variation of the methane yield during the start-up period of an anaerobic fluidized bed reactor. After a lag phase, with acclimatized sludge, the methane yield increased with time during biofilm development up to the theoretical steady yield value, reported to be around 0.351 CH4/g CODdeg. The establishment of the biofilm required a high consumption of organic material through the microbial synthesis (anabolism), thereby reducing the proportion of substrate converted to methane. As a result, this yield could be an indirect metabolic parameter for evaluating a start-up operation. It could provide vital information about bacterial fixation processes and is easy to be applied to any biofilm reactor, such as anaerobic filters, where biomass sampling is impracticable. Monitoring this parameter could also give useful dynamic information about the different steps of colonization and biomass attachment, which could be used to improve the start-up performance. PMID- 11902796 TI - Recent trends in stabilizing protein structure upon encapsulation and release from bioerodible polymers. AB - Sustained release of pharmaceutical proteins from biocompatible polymers offers new opportunities in the treatment and prevention of disease. The manufacturing of such sustained-release dosage forms, and also the release from them, can impose substantial stresses on the chemical integrity and native, three dimensional structure of proteins. Recently, novel strategies have been developed towards elucidation and amelioration of these stresses. Non-invasive technologies have been implemented to investigate the complex destabilization pathways that can occur. Such insights allow for rational approaches to protect proteins upon encapsulation and release from bioerodible systems. Stabilization of proteins when utilizing the most commonly employed procedure, the water-in-oil-in-water (w/o/w) double emulsion technique, requires approaches that are based mainly on either increasing the thermodynamic stability of the protein or preventing contact of the protein with the destabilizing agent (e.g. the water/oil interface) by use of various additives. However, protein stability is still often problematic when using the w/o/w technique, and thus alternative methods have become increasingly popular. These methods, such as the solid-in-oil-in-oil (s/o/o) and solid-in-oil-in-water (s/o/w) techniques, are based on the suspension of dry protein powders in an anhydrous organic solvent. It has become apparent that protein structure in the organic phase is stabilized because the protein is "rigidified" and therefore unfolding and large protein structural perturbations are kinetically prohibited. This review focuses on strategies leading to the stabilization of protein structure when employing these different encapsulation procedures. PMID- 11902797 TI - Poly(L-lysine) as a model drug macromolecule with which to investigate secondary structure and membrane transport, part I: Physicochemical and stability studies. AB - Low oral bioavailability of therapeutic peptides and proteins generally results from their poor permeability through biological membranes and enzymatic degradation in the gastrointestinal tract. Since different secondary structures exhibit different physicochemical properties such as hydrophobicity, size and shape, changing the secondary structure of a therapeutic polypeptide may be another approach to increasing its membrane permeation. Poly(L-lysine) was used as a model polypeptide. The objectives of this study were to induce secondary structural changes in poly(L-lysine) and to determine the time course over which a given conformer was retained. In addition, the hydrophobicity of each secondary structure of poly(L-lysine) was assessed. The circular dichroism (CD) studies demonstrated that the conditions employed could successfully induce the desired secondary structural changes in poly(L-lysine). The alpha-helix conformer appeared to be more stable at 25 degrees C whereas the beta-sheet conformer could be preserved at 37 degrees C. On the other hand, the random coil conformer was retained at both temperatures. Significant losses of the alpha-helix and the beta sheet conformers were observed when the pH was reduced. The change in ionic strength did not affect any of the conformers. The octanol/buffer partitioning studies indicated that the alpha-helix and the beta-sheet conformers exhibited significantly different (P < 0.05) hydrophobicities. In conclusion, variation of pH and temperature conditions can be used to induce secondary structural changes in poly(L-lysine). These changes are reversible when the stimuli are removed. The alpha-helix and the beta-sheet conformers of poly(L-lysine) are more lipophilic than the native random coil conformer. Thus, poly(L-lysine) may represent an ideal model polypeptide with which to further investigate the effects of secondary structure on membrane diffusion or permeation. PMID- 11902798 TI - Crystallization conditions and formation of orthorhombic paracetamol from ethanolic solution. AB - Orthorhombic paracetamol exhibits far better tabletability than the monoclinic form and its bulk crystallization from solution attracts much interest. In this study, temperature changes in supersaturated ethanolic solution have been recorded after seeding with orthorhombic crystals under different cooling temperatures (Tc) and agitation rates (AR). Average cooling rate (CR), time for maximum temperature deviation (tmax) and area confined between curves of measured and reference temperature plots (AUC) were calculated and correlated with crystal yield (Y). The micromeritic (size and shape) and the compression properties, the density and the orthorhombic content of the crystalline product were evaluated and related to the main crystallization conditions applied (Tc and AR). Conditions for optimal crystal yield and orthorhombic content were elucidated. It was found that crystal yield (Y) increased with AR and decreased with Tc. The ratio tmax/CR provided good prediction of crystal yield (Y = 58.92-1.386 tmax/CR, r2 = 0.964 and P = 0.0001). Tc and AR linearly affected crystal size and the size distribution, probably due to alterations in supersaturation, but they did not affect the crystal shape significantly. Density and compression properties (yield pressure and elastic recovery) were determined by the content of the orthorhombic form, which increased linearly with AR (P = 0.009) and with Tc (P = 0.039) when agitation was between 300 and 500 rev min(-1), while tmax decreased. At 700 rev min(-1) orthorhombic content was maximized and became independent to Tc. Higher orthorhombic content and crystal yield was expected for lower Tc and for lower tmax, which corresponded to higher AR and might have also been affected by alteration of seeding and harvesting procedure. PMID- 11902799 TI - Stereoselective distribution and stereoconversion of zopiclone enantiomers in plasma and brain tissues in rats. AB - Concentrations of (-)-zopiclone and (+)-zopiclone were determined in plasma and brain after oral administration, to investigate the stereoselectivity of distribution in rats. Zopiclone enantiomers were administered separately to rats and concentrations were determined by chiral HPLC in plasma and brain. In initial experiments, rats were treated with urethane before cannulation for blood sampling but as this drug modified zopiclone pharmacokinetics, it was not used in subsequent studies. This study showed that zopiclone pharmacokinetics after oral gavage in rats are stereoselective. After oral administration of (+)-zopiclone, no stereoconversion was observed in plasma. Conversely, after administration of ( )-zopiclone, both enantiomers were found in plasma and brain with (+)-zopiclone/( )-zopiclone ratios of 1 and 8.4 in plasma and brain, respectively. Our findings suggest that zopiclone undergoes stereoconversion and that it is stereospecifically distributed to the brain. PMID- 11902800 TI - Omeprazole increases permeability across isolated rat gastric mucosa pre-treated with an acid secretagogue. AB - Triple therapy using proton-pump inhibitors (PPIs) in combination with oral antibiotics for the treatment of Helicobacter pylori-associated gastritis has shown increased efficacy for reasons that are still poorly understood. Possible explanations include a direct antibacterial effect of the PPIs or a PPI-mediated increase in bacterial susceptibility to antibiotics. Using an in-vitro model of rat gastric mucosa, we examined fluxes of a radiolabelled marker molecule through the interepithelial tight junctions under normal conditions and under the influence of an acid secretagogue (50 microM histamine) and a PPI (100 microM omeprazole). Paracellular fluxes of the radiolabel (represented by calculation of apparent permeability coefficients) were linear over 2 h. Fluxes of the marker increased significantly after treatment with histamine followed by omeprazole, but were unaltered in paired preparations exposed to the same drugs given in reverse order. Enhancements in paracellular permeability were mirrored in separate experiments using a detergent (Triton X-100), a bile salt (deoxycholate) and an agent that disrupts the cytoskeleton (cytochalasin D) to interfere with tight junctional integrity. The results suggest that exposure of acid-secreting gastric mucosa to omeprazole widens the interepithelial spacing in a manner that may facilitate enhanced macromolecular transport. Increases in antibiotic delivery from the blood to the gastric lumen via such a mechanism may account for the greater eradication rates observed with PPI-based triple therapy in H. pylori associated gastritis. PMID- 11902801 TI - Synthesis and physicochemical assessment of novel 2-substituted 3-hydroxypyridin 4-ones, novel iron chelators. AB - Novel 3-hydroxypyridin-4-one containing tridentate ligands were synthesised and their physicochemical properties characterised, including ionisation constants and stoichiometric titration with Fe(III). There is an urgent demand for orally active iron chelators with potential for the treatment of thalassaemia. In principle, tridentate ligands are likely to be more kinetically stable than bidentate molecules, but to date no satisfactory molecules have been identified. Fe(III) stability constants were assessed by competition with the hexadentate ligand EDTA. In all cases no evidence was found for a tridentate mode of iron chelation; instead the ligands behaved as bidentate hydroxypyridinones. As a consequence they provide no advantage over the more simple alkyl hydroxypyridinones. PMID- 11902802 TI - Anti-inflammatory and antioxidant properties of Helichrysum italicum. AB - The anti-inflammatory and antioxidant activities of the aerial part of Helichrysum italicum extracts have been established in various in-vivo and in vitro experimental models. The results obtained on the acute oedemas induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) and ethyl phenylpropiolate in the mouse ear, by serotonin and phospholipase A2 (PLA2) in the mouse paw, on chronic inflammation induced by repeated application of TPA in the mouse ear and on the delayed-type hypersensitivity induced by sheep red blood cells suggest that said anti-inflammatory activity is due to the effects of compounds expressed via a corticoid-like mechanism. In addition, the antioxidant activity of the extracts seems to be implicated in this anti-inflammatory activity, as the former inhibits enzymatic and non-enzymatic lipid peroxidation and has free-radical scavenger properties. We conclude that the anti-inflammatory activity of Helichrysum italicum can be explained by multiple effects, including inflammatory enzyme inhibition, free-radical scavenging activity and corticoid-like effects. PMID- 11902804 TI - 1-O-hexyl-2,3,5-trimethylhydroquinone inhibits IkappaB phosphorylation and degradation-linked inducible nitric oxide synthase expression: beyond antioxidant function. AB - Inducible nitric oxide (NO) production in macrophages plays an important role in atherosclerosis, the protective effects of vitamin E and its derivatives perhaps being partly mediated by alteration in this parameter. We have investigated the influence of a novel synthesized vitamin E derivative, 1-O-hexyl-2,3,5 trimethylhydroquinone (HTHQ), on NO production in the RAW 264.7 mouse macrophage cell line. HTHQ dose-dependently inhibited lipopolysaccharide (LPS)-induced NO production through reducing LPS-triggered inducible nitric oxide synthase (iNOS) expression. The phosphorylation and subsequent degradation of IkappaB caused by LPS in RAW 264.7 cells was markedly blocked. The free radical scavenging activity of HTHQ was only 2-fold that of vitamin E, whereas its inhibition of NO production was found to be nearly 500-fold stronger. Our results indicated that HTHQ suppressed NO production in macrophages by blocking IkappaB degradation and thus inhibiting iNOS expression. The inhibitory activity of HTHQ on NO production did not parallel its free radical scavenging activity, implying a possible involvement of additional functions. PMID- 11902803 TI - Distribution kinetics of solutes in the isolated in-situ perfused rat head using the multiple indicator dilution technique and a physiological two-barrier model. AB - The purpose of this study was to determine the pharmacokinetics of [14C]diclofenac, [14C]salicylate and [3H]clonidine using a single pass rat head perfusion preparation. The head was perfused with 3-[N-morpholino] propane sulfonic acid-buffered Ringer's solution. 99mTc-red blood cells and a drug were injected in a bolus into the internal carotid artery and collected from the posterior facial vein over 28 min. A two-barrier stochastic organ model was used to estimate the statistical moments of the solutes. Plasma, interstitial and cellular distribution volumes for the solutes ranged from 1.0 mL (diclofenac) to 1.6 mL (salicylate), 2.0 mL (diclofenac) to 4.2 mL (water) and 3.9 mL (salicylate) to 20.9 mL (diclofenac), respectively. A comparison of these volumes to water indicated some exclusion of the drugs from the interstitial space and salicylate from the cellular space. Permeability-surface area (PS) products calculated from plasma to interstitial fluid permeation clearances (CL(PI)) (range 0.02-0.40 mL s(-1)) and fractions of solute unbound in the perfusate were in the order: diclofenac > salicylate > clonidine > sucrose (from 41.8 to 0.10 mL s(-1)). The slow efflux of diclofenac, compared with clonidine and salicylate, may be related to its low average unbound fraction in the cells. This work accounts for the tail of disposition curves in describing pharmacokinetics in the head. PMID- 11902805 TI - Hyperhomocysteinaemia and cardiovascular risk in female ovariectomized rats: role of folic acid and hormone replacement therapy. AB - Hyperhomocysteinaemia is an independent risk factor for arteriosclerosis, recurrent thromboembolic complications and osteoporosis. After menopause, a high level of total homocysteine seems to be secondary to the altered hormonal status. Hormone replacement therapy (HRT) limits the development of coronary artery disease through a variety of mechanisms. One such mechanism is through affecting homocysteine metabolism. Folate and vitamin B12 deficiencies are considered to be major risks for hyperhomocysteinaemia. This study, therefore, was undertaken to examine whether lowering homocysteine with HRT or folic acid in ovariectomized rats could attenuate cardiovascular complications. Sixty sexually mature female Wistar rats were ovariectomized. Three weeks later, they were treated with estradiol (15 microg kg(-1), every two weeks, i.m.) or folic acid (90 microg daily, orally), either alone or in a combined form for four weeks. In addition, groups of ovariectomized rats (positive control) and healthy rats (negative control) were given cottonseed oil. Blood samples were then collected for serum and plasma separation. Serum total homocysteine, folate, estradiol, plasma nitric oxide (NO), lipid profile, and susceptibility of non-high-density-lipoprotein cholesterol (non HDLC) content to oxidation were determined. In ovariectomized rats, hyperhomocysteinaemia was established and associated with significant increments of both atherogenic indexes (total cholesterol/HDLC, low-density lipoprotein cholesterol (LDLC)/HDLC) and susceptibility of their non HDLC to oxidation. However, plasma NO, serum folate, and estradiol levels significantly decreased. HRT and folic acid significantly reduced total homocysteine and susceptibility of non HDLC to oxidation and increased plasma NO content. Moreover, a significant negative correlation was found between total homocysteine versus folate and estradiol (r = -0.5, P < 0.01; r = -0.25, P < 0.05, respectively). Meanwhile, a positive correlation with the susceptibility of lipoprotein to oxidation was observed (r = 0.85, P < 0.001). In conclusion, a low folate level is found to be associated with elevated total homocysteine. Folic acid supplementation, either individually or in a combined form with HRT, has a beneficial effect in low estrogen status subsequent to ovariectomy. PMID- 11902806 TI - Gastroprotective activity of solidagenone on experimentally-induced gastric lesions in rats. AB - The gastroprotective effect of the labdane diterpene solidagenone was assessed on gastric ulcer in rats. The effect of a single oral dose of the compound was evaluated at 50, 100 and 200 mg kg(-1) in the following test systems: pylorus ligature (Shay), aspirin- and ethanol-induced gastric ulcers. In pylorus-ligated rats (Shay model), the ulcerative index decreased by 37% with solidagenone pre treatment at the three assayed doses. The effect of a single oral dose of 50 mg kg(-1) solidagenone was comparable with ranitidine at the same concentration and similar to higher doses of the compound. A significant effect (P < 0.001) at 100 and 200 mg kg(-1) was observed in the aspirin-induced ulcer model. At both doses, reduction in the number of lesions was approximately 50% compared with controls. The effect was comparable with the reference compound ranitidine (50 mg kg(-1)). With the ethanol-induced gastric ulcers, the effect of solidagenone at 100 and 200 mg kg(-1) was similar to a single oral dose of 20 mg kg(-1) omeprazole with a 50% reduction of the mean number of lesions compared with controls. In acute toxicity tests on mice, intraperitoneal administration of solidagenone showed no toxicity at doses up to 600 mg kg(-1). This is the first report on the gastroprotective activity of a labdane diterpene. PMID- 11902807 TI - Involvement of peripheral cyclooxygenase-1 and cyclooxygenase-2 in inflammatory pain. AB - Pain-induced functional impairment in the rat (PIFIR) is a model of inflammatory and arthritic pain similar to that of clinical gout. Nociception is induced by the intra-articular injection of uric acid into the right hind limb, inducing its dysfunction. Animals then receive analgesic drugs and the recovery of functionality over time is assessed as an expression of antinociception. We have examined the role of peripheral prostaglandins synthesized by cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in inflammatory pain using the PIFIR model. Rofecoxib (a selective COX-2 inhibitor) and SC-560 (a selective COX-1 inhibitor) both produced dose-dependent effects. When the inhibitors were administered before uric acid, they showed similar potency, but the antinociceptive efficacy of SC-560 was lower than rofecoxib; the best antinociceptive effects were obtained with the dose of 100 microg/articulation of each inhibitor (pre treatment). In post-treatment (inhibitors administered after the uric acid), rofecoxib showed the least antinociceptive effect and SC-560 was more potent than rofecoxib. The inhibition of both COX-1 and COX-2 produced a more profound analgesic effect than the inhibition of either COX-1 or COX-2 alone. The present data support the idea that both COX isoforms contribute to the development and maintenance of local inflammatory nociception. Thus, it could be expected that inhibition of both COX-1 and COX-2 is required for non-steroidal anti inflammatory drugs (NSAID)-induced antinociception in the rat. These findings suggest that the therapeutic effects of NSAIDs may involve, at least in part, inhibition of COX-1 and COX-2. PMID- 11902808 TI - The long-lasting anti-anginal effects of CP-060S in a rat model of arginine vasopressin-induced myocardial ischaemia. AB - The anti-anginal effect of CP-060S, a new cardioprotective agent that prevents myocardial Na+-, Ca2+-overload and has Ca2+-channel blocking activity, was evaluated in a rat model of arginine8-vasopressin (AVP)-induced cardiac ischaemia. Infusion of AVP (0.2 IU kg(-1)) depressed the electrocardiogram (ECG) ST segment, an index of myocardial ischaemia. Vehicle, CP-060S and diltiazem were given orally 1, 2, 4, 8, 12 and 24 h before the administration of AVP. CP-060S, at 3 mg kg(-1) and 10 mg kg(-1), suppressed AVP-induced ST-segment depression for 2 h and 12 h, respectively. In contrast, diltiazem, at 10 and 30 mg kg(-1), suppressed AVP-induced ST-segment depression for only 1 h. The persistent suppression of the AVP-induced ST-segment depression by CP-060S correlated with the time course of changes in its plasma concentration. The minimum effective concentration of CP-060S was estimated to be 30 ng mL(-1) (approximately 50 nM), consistent with its vasorelaxant potency in rat isolated aortic strips (concentration producing 50% relaxation of KCl contraction, IC50 = 32.6+/-8.3 nM). Intravenously administered CP-060S, at 300 microg kg(-1) and diltiazem at 500 microg kg(-1) showed similar haemodynamic changes, whereas CP-060S, at 300 microg kg(-1), significantly suppressed AVP-induced ST-segment depression and diltiazem, at 500 microg kg(-1), had no effect on AVP-induced ST-segment depression. In summary, orally administered CP-060S exerted a long-lasting anti anginal effect proportionate to the time course of changes in its plasma concentration in a rat model of AVP-induced ischaemia. PMID- 11902809 TI - Simvastatin and lovastatin, but not pravastatin, interact with MDR1. AB - The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, pravastatin, was compared with simvastatin and lovastatin from the viewpoint of susceptibility to interaction with or via the multidrug transporter, MDR1 (P glycoprotein). This was carried out using the MDR1-overexpressing cell line LLC GA5-COL150, established by transfection of MDR1 cDNA into porcine kidney epithelial LLC-PK1 cells, and [3H]digoxin, which is a well-documented substrate for MDR1. Pravastatin, at 25-100 microM, had no effect on the transcellular transport of [3H]digoxin whereas simvastatin and lovastatin suppressed the basal to-apical transport of [3H]digoxin and increased the apical-to-basal transport. It was suggested that recognition by MDR1 was due to the hydrophobicity. In conclusion, simvastatin and lovastatin are susceptible to interaction with or via MDR1, but pravastatin is not. This is important information when selecting the HMG-CoA reductase inhibitors for patients taking drugs that are MDR1 substrates. PMID- 11902810 TI - Characterisation of the effects of potassium channel modulating agents on mouse intestinal smooth muscle. AB - The actions of agents which modulate ATP-sensitive potassium (K(ATP)) channels in excitable cells were investigated in an in-vitro preparation of mouse ileum from which the mucosa was removed. A range of potassium channel openers of diverse structure, cromakalim (0.1-100 microM), pinacidil (0.1-200 microM) and its analogue P1060 (0.1-200 microM), SDZ PCO400 ((-)-(3S,4R)-3,4-dihydro-3-hydroxy 2,2-dimethyl-4-(3-oxo-cyclopent-1-enyloxy)-2H-1-benzopyran-6-carbonitrile) (0.3 60 microM), caused concentration-related reduction in twitch height of electrical field stimulated ileum. P1060 and SDZ PCO400 were the most potent agents; diazoxide (0.1-100 microM) was without effect. The order of inhibitory potency, based on EC50 values (concentration of a relaxant producing 50% of the maximum inhibition of twitch) was: P1060 = SDZ PCO400 > cromakalim > pinacidil. The relaxant effect of the potassium channel openers was antagonised by the sulfonylureas glibenclamide (0.1-1.0 microM) and glipizide (3-30 microM) but the nature of the antagonism differed. Antagonism of P1060 and SDZ PCO400 by glibenclamide appeared to be competitive whereas the antagonism of relaxation induced by cromakalim and pinacidil was apparently not competitive. Both phentolamine (1-10 microM) and tolbutamide (100-300 microM) showed competitive antagonism of the actions of pinacidil while yohimbine (1-20 microM) did not antagonise relaxation and appeared to have actions at sites other than the K(ATP) channel in this preparation. The relative effectiveness of the antagonists on pinacidil-induced relaxation was found to be: glibenclamide > phentolamine > tolbutamide > yohimbine, which is in agreement with studies in other tissues. The results show that many structurally diverse potassium channel openers are potent relaxants of mouse ileum. These observations are consistent with the existence of ATP-dependent K+ channels in murine intestinal muscle which, however, differ somewhat in properties from those reported for vascular muscle and pancreatic beta-cells. PMID- 11902811 TI - A validated high performance liquid chromatographic method for the analysis of Goldenseal. AB - Goldenseal (Hydrastis canadensis L.) has emerged as one of the top ten herbal supplements on the worldwide market. A rapid, simple and validated high performance liquid chromatographic method, with photodiode array detection, has been developed for the analysis of commercial Goldenseal products. Samples were treated by sonication with acidified methanol/water. The method was validated for LOD, LOQ, linearity, reproducibility and recovery with good results. PMID- 11902812 TI - Sensitivity of dissolution rate to location in the paddle dissolution apparatus. AB - The aim of the present study was to determine the apparent diffusion boundary layer and dissolution rate constant for various surfaces of compacts and at various locations in the USP paddle dissolution apparatus. Benzoic acid compacts were coated with paraffin wax leaving only the surface under investigation free for dissolution. The dissolution rates for various surfaces at varying locations in the paddle dissolution vessel were determined from the slope of the dissolution profile (amount dissolved (mg) versus time (min)). The apparent diffusion boundary layer and dissolution rate constant were calculated and were found to vary depending on the surface of the compact from which dissolution took place and also on the location and size of the compact. It may be concluded that, in developing models to describe the dissolution from solid dosage forms, it is not accurate to assume constant hydrodynamics and mass transfer rates at all surfaces of the system, or in different locations within the test device. A more exact description of the hydrodynamics would be necessary in order to precisely model drug dissolution in the paddle dissolution apparatus. PMID- 11902813 TI - The study on organic nitrates, part V. New derivatives of piperazine potential NO donors. AB - We have obtained a series of non-symmetrical 1,4-disubstituted derivatives of piperazine, with the structure of organic nitrates, as potential NO donors. These compounds were obtained from respective hydroxyl derivatives of piperazine in an esterification reaction by fuming nitric acid. The obtained nitrates were tested in-vitro by reaction with a sulfhydryl compound. The structure of the most active nitrate and its hydroxyl analogue was used for the calculation of geometrical optimization with the determination of 3D-QSAR by a semi-empirical method PM3 using HyperChem 4.5. PMID- 11902814 TI - Snuff--how dangerous is it? The controversy continues. PMID- 11902816 TI - Acute and chronic neutropenias. What is new? AB - Recently, some of the mechanisms and consequences in the severe chronic neutropenias (e.g. the neutrophil elastase gene mutations and the risk to progress to myelodysplasia and acute leukaemia) and in drug-induced agranulocytosis (e.g. the apoptosis-inducing ability of metabolites of clozapine) have been elucidated, and new aspects of autoimmune and the large granular lymphocyte syndrome were described (e.g. aberrant elaboration of Fas-ligand causing neutrophil apoptosis). Investigations of the mild to moderate chronic neutropenias have shown the significance of interactions between the myeloid development and the immune network (e.g. relations to immunoglobulin aberrations). Granulocyte-colony stimulation factor (G-CSF) is widely used in patients with severe chronic neutropenia, however, its use in other conditions is mostly based on anecdotal evidence. In addition, immune modulating regimens, such as metothrexate, ciclosporine and monoclonal antibodies, are increasingly employed for the autoimmune neutropenias. PMID- 11902815 TI - Progress in the development of immunotherapy for the treatment of patients with cancer. AB - Several recent developments have hallmarked progress in tumour immunology and immunotherapy. The use of interleukin-2 (IL-2) in cancer patients demonstrated that an immunological manipulation was capable of mediating the regression of established growing cancers in humans. The identification of the genes encoding cancer antigens and the development of means for effectively immunizing patients against these antigens has opened important new avenues of exploration for the development of effective active and cell-transfer immunotherapies for patients with cancer. PMID- 11902817 TI - Carotid and femoral atherosclerosis, cardiovascular risk factors and C-reactive protein in relation to smokeless tobacco use or smoking in 58-year-old men. AB - Objectives. To examine the associations between smokeless tobacco use, smoking, cardiovascular risk factors, inflammation and ultrasound-assessed measures of atherosclerosis in the carotid and femoral arteries. Subjects. The study was performed in a population-based sample of clinically healthy men (n = 391) all 58 years old. Exclusion criteria were cardiovascular or other clinically overt diseases or continuous medication with cardiovascular drugs. Methods. The habits of smoking and oral moist snuff use were assessed by questionnaires. C-reactive protein (CRP) was assessed by high sensitive enzyme-linked immunosorbent assay (ELISA). Intima-media thickness (IMT) in the carotid bulb, the common carotid artery and the common femoral artery and plaque occurrence were measured by ultrasound. Results. The use of oral moist snuff was associated with serum triglycerides and waist-hip ratio (WHR), but not with CRP or ultrasound-assessed measures of subclinical atherosclerosis. Smoking, on the other hand, was associated with CRP, the components in the metabolic syndrome and IMT as well as plaques in the carotid and femoral arteries. In comparison to never-smokers the current smokers had higher values of WHR, triglycerides, C-reactive protein and IMT in carotid bulb and femoral artery. Ex-smokers were in general more obese and had a femoral IMT that was in-between that of never-smokers and current smokers. Conclusions. Tobacco smoking, but not oral moist snuff use, was associated with carotid and femoral artery IMT, and increased levels of CRP. Current smoking was also associated with abdominal obesity. Ex-smokers though, are generally more obese. Smoking was also associated with hyperinsulinaemia, dyslipidaemia and high blood pressure, i.e. the metabolic syndrome. The inhaled smoke from the combustion of tobacco seems to be an important aetiological factor in the atherosclerotic process. PMID- 11902818 TI - Urinary albumin excretion is independently associated with C-reactive protein levels in overweight and obese nondiabetic premenopausal women. AB - OBJECTIVES: C-reactive protein (CRP) and microalbuminuria are nowadays considered markers of chronic inflammation of the arterial wall and of endothelial dysfunction, respectively. An increase of CRP levels and of urinary albumin excretion (UAE) rate have both been reported to be independently associated with a higher risk of cardiovascular morbidity and mortality in the general population. The aim of the present study was to evaluate the possible correlation between UAE and CRP concentrations in overweight and obese premenopausal women. DESIGN AND SETTING: A cross-sectional study in a primary care setting. SUBJECTS, MAIN OUTCOME MEASURES: CRP levels and UAE rate were measured in 103 overweight and obese premenopausal women, aged 18-45 years. Other measurements included: central fat accumulation, as evaluated by waist circumference, insulin resistance, as calculated by homeostatic model assessment (HOMAIR); fat-free mass (FFM), as measured by bioimpedance analysis; blood pressure; and fasting plasma levels of glucose, insulin, and lipids. RESULTS: Urinary albumin excretion was positively correlated with body mass index (BMI) (P < 0.01), waist circumference (P < 0.001), diastolic blood pressure (P < 0.01), triglycerides (P < 0.01), HOMAIR (P < 0.05), and CRP levels (P < 0.05); and negatively associated with HDL cholesterol (P < 0.001). After multivariate analysis, diastolic blood pressure, HDL cholesterol, and CRP levels maintained their significant correlation with UAE. CONCLUSION: Our study shows a strong relationship between UAE and CRP concentrations, irrespective of age and other anthropometric and metabolic variables. On this basis, it can be argued that inflammation of the arterial wall, as indicated by higher CRP plasma levels, and endothelial dysfunction, as shown by higher UAE rate, might represent simultaneous phenomena in the development of atherosclerosis in overweight and obese premenopausal women. PMID- 11902820 TI - The NeiI polymorphism in the cyclin D1 gene and sporadic primary hyperparathyroidism. AB - OBJECTIVES: The cell cycle regulator cyclin D1 plays an important role in parathyroid tumourigenesis. The NciI polymorphism in exon 4 has recently been associated with early onset of hereditary nonpolyposis colorectal cancer and is a prognostic indicator of nonsmall cell lung cancer and squamous cell carcinomas. Furthermore, a limited study of 28 primary hyperparathyroidism (pHPT) patients displayed a tendency of NciI influence on HPT development. We hypothesized that the NciI polymorphism may relate to a risk of developing pHPT. DESIGN, SETTING AND SUBJECTS: We genotyped 182 patients with sporadic pHPT and matched controls for the cyclin D1 polymorphism. A total of 88 pHPT patients and controls were recruited via a health-screening. RESULTS: The frequency distribution of the NciI genotypes NN, Ni, and ii were in pHPT patients and controls 22, 44 and 34%, and 26, 49 and 25%, respectively. The calculated allele frequencies were A = 0.56; G = 0.44 in cases and A = 0.49; G = 0.51 in controls. The frequency distributions did not differ comparing cases and controls when subgrouped after age and menopausal status. The NciI genotypes were not significantly associated with age of the individuals, serum (s)-calcium, s-parathyroid hormone (PTH), bone mineral density (BMD) or parathyroid tumour weight in any of the groups of pHPT patients or controls. CONCLUSIONS: No significant differences in distribution of the genotypes could be detected between the groups, suggesting that the polymorphism has minor or no pathogenic importance in the development of pHPT. Our results suggest that determination of the NciI polymorphism in the cyclin D1 gene is not a clinically useful tool for prediction of pHPT. PMID- 11902819 TI - Combined hormone replacement therapy improves endothelial function in healthy postmenopausal women. AB - OBJECTIVES: Large scale epidemiological studies suggest that hormone replacement therapy (HRT) reduces cardiovascular events in postmenopausal women. Improvement in endothelial function may contribute to this protective effect. DESIGN: In a prospective, double blind study, 61 healthy postmenopausal women were randomized to receive either oral continuous combined HRT [oestradiol 2 mg and norethisterone acetate (NETA) 1 mg per day] or placebo. Endothelial function, assessed by flow-mediated vasodilation (FMD) of the brachial artery and expression of soluble endothelial cell adhesion molecules (CAM) were determined before, after 3 and 6 months of therapy. RESULTS: The FMD was significantly improved in women on combined HRT (from 5.97% to 10.94% after 3 months and to 10.58% after 6 months; both P < 0.01 versus baseline values) and did not change in the placebo group (6.92% at baseline, 5.86% after 3 and 6.26% after 6 months). After 3 months of combined HRT, significant decreases of 24.6% for E-selectin and 13.9% for intercellular adhesion molecule-1 (ICAM-1) were observed (both P < 0.01 versus baseline values) and were sustained after 6 months of therapy, whilst no differences emerged in the placebo group. CONCLUSIONS: Oestradiol and norethisterone acetate improve endothelial function by both enhancing FMD and reducing the levels of soluble E-selectin and ICAM-1 in healthy postmenopausal women. PMID- 11902821 TI - Sleep disturbance in association with elevated pulse rate for prediction of mortality--consequences of mental strain? AB - OBJECTIVES: Sleep deprivation has experimentally been shown to adversely influence glucose metabolism, endocrine function and sympathovagal balance in young men without known serious disease. We investigated the impact of sleep problems and resting heart rate in a large sample of self-reported, healthy middle-aged men and women on long-term mortality. METHODS: In all 22,444 men and 10,902 women participated in a population-based health screening (71% mean attendance), including blood sampling and examination of blood pressure (BP) and pulse rate after 10 min supine rest, as well as a self-administered questionnaire on sleep problems. Mortality was assessed from national death registers. RESULTS: Sleep disturbances were related to increased cardiovascular risk factor levels at baseline in both sexes, and predicted total and cause-specific mortality after a mean of 12 years (women) and 17 years (men) of follow-up. In men, self-reported healthy at baseline, total mortality during follow-up was independently predicted by both sleep problems and increased resting heart rate, also after adjustment for smoking, body mass index (BMI), systolic BP, cholesterol, smoking and problematic alcohol drinking habits. A step-wise increased total mortality was shown in men reporting successively worse sleep problems and higher heart rate, highest hazard ratio 2.7 [95% confidence interval (CI) = 2.1-3.4] after adjustments, compared with men free from sleep problems and with normal heart rate. CONCLUSIONS: Sleep disturbance is a predictor of total and cause-specific mortality in both sexes, but only interacts with increased resting heart rate for this prediction in healthy men. Sleep problems correlated cross-sectional with disturbances in lipid and glucose metabolism, even after adjustment for degree of obesity and smoking. Sleep disturbance is a symptom for a biological pathway that is correlated to premature mortality. One possible explanation would be that it acts in concert with sympathetic nervous activation (SNA), both being consequences of chronic stress exposure. PMID- 11902822 TI - Adrenocorticotrophic hormone exerts marked lipid-lowering effects in simvastatin treated patients. AB - OBJECTIVE: Recently, it was reported that treatment with adrenocorticotrophic hormone (ACTH) has a strong lipid-lowering effect in healthy individuals. The mechanism behind this has not been established. The aim of the present investigation was to study the effect of ACTH on the plasma lipoprotein pattern in patients treated with a 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor. DESIGN: The ACTH treatment was given to 10 patients who were on long term treatment with simvastatin 40 mg daily. ACTHI-24 was administered at the dose of 1 mg daily for four consecutive days. Blood samples for analyses of lipids, lipoproteins and apolipoproteins were collected before and after treatment. Second baseline was obtained 2 weeks after the end of treatment. RESULTS: The serum concentrations of cholesterol, triglycerides, low density lipoprotein (LDL) cholesterol, apolipoprotein B and lipoprotein(a) fell significantly by 16, 23, 23, 10 and 38%, respectively. The serum apolipoprotein E concentration increased significantly by 39%; the fraction that was not associated with apolipoprotein B increased by 47% whereas the fraction that was did not change significantly. There were no changes in the serum concentrations of high density lipoprotein (HDL) cholesterol and apolipoprotein AI. At the second baseline, the lipid variables had generally returned to previous levels. CONCLUSIONS: In patients on long-term simvastatin treatment, ACTH had marked lowering effects on the lipoproteins that contain apolipoprotein B. Moreover, the serum apolipoprotein E concentration increased significantly in response to ACTH treatment. PMID- 11902823 TI - Specific bronchoalveolar lavage fluid T cells associate with disease in a pair of monozygotic twins discordant for sarcoidosis. AB - A 49-year-old Caucasian woman had an acute onset of sarcoidosis. Bronchoscopy with bronchoalveolar lavage (BAL) showed a pronounced accumulation of BAL fluid CD4+ T cells expressing the T-cell receptor (TCR) AV2S 3 gene. In line with this observation, the patient was HLA-DR 17 positive, previously shown to strongly correlate with lung compartmentalized AV2S3+ T cells. At follow-up after recovery, reduced numbers of BAL fluid AV2S3+ T cells were found. Interestingly, BAL fluid of a healthy monozygotic twin sister contained normal numbers of AV2S3+ lung T cells. This report shows the T-cell repertoire of BAL fluid T cells to correlate with the disease (sarcoidosis). indicating a local and specific immune response triggered by an unknown antigen in sarcoidosis. PMID- 11902824 TI - Fas/FasL-independent activation-induced cell death of T lymphocytes from HIV infected individuals occurs without DNA fragmentation. AB - We assessed the effects of activation with phorbol myrystic acetate (PMA) and ionomycin on peripheral blood mononuclear cells (PBMC) from HIV-infected individuals by (51)Cr release, propidium iodide (PI) uptake, electron microscopy, and DNA analysis. Up to 70% (51)Cr release was induced from PBMC of HIV-infected individuals, versus up to 26% (51)Cr release from PBMC of non-HIV-infected volunteers. Flow cytometry identified mostly T cells undergoing activation induced cell death (AICD). The kinetics of (51)Cr release and the effects of cold target inhibitors were consistent with cell-mediated cytotoxicity. Certain anti CD3 antibodies or extracellular Ca(2+) chelation prevented AICD, but antagonistic anti-Fas antibodies, caspase inhibitors, and cycloheximide had no effect. The antioxidants thiourea and N-acetylcysteine reduced AICD, indicating a role for oxidative stress. Electron microscopy revealed plasma membrane disruption with nuclear integrity, while DNA analysis showed intact chromosomal DNA. This form of T cell AICD triggered by PMA and ionomycin differs from classical apoptosis in the absence of either caspase involvement or DNA fragmentation. PMID- 11902825 TI - Natural killer cells express estrogen receptor-alpha and estrogen receptor-beta and can respond to estrogen via a non-estrogen receptor-alpha-mediated pathway. AB - Natural killer (NK) cells play a crucial role in host defense against pathogens and immune surveillance against cancer. Given that estrogens have been reported to suppress NK cell activity, we sought to elucidate the mechanisms by which estrogen mediates this effect. We demonstrate by immunocytochemical staining with estrogen receptor-alpha (ERalpha)- and estrogen receptor-beta (ERbeta)-specific antibodies that both ERalpha and ERbeta are expressed in murine NK cells. We also compared the ability of high doses of 17beta-estradiol ( approximately 800 pg/ml) to regulate NK cell activity in wild-type and estrogen receptor-alpha-deficient (ERalphaKO) mice. 17beta-estradiol elicited a significant decrease in NK cell activity in both wild-type and ERalphaKO mice (P < 0.001). These data suggest that ERbeta or possibly a novel receptor is involved in mediating estrogen action on NK cell activity and raise the potential for therapeutic modulation of NK cell activity with selective estrogen receptor modulators (SERMS). PMID- 11902826 TI - Intercellular exchange of class II MHC complexes: ultrastructural localization and functional presentation of adsorbed I-A/peptide complexes. AB - Activated rat T cells, like human T cells, synthesize class II MHC glycoproteins (MHCII) and absorb MHCII from neighboring T cells. This study focused on interactions of myelin basic protein (MBP)-specific T cells that either synthesized MHCII or absorbed MHCII during activation to assess cellular structures associated with presentation of functional MHCII/peptide complexes. Synthesis of MHCII by CD4(+)TCR(+) T cells involved I-A(+) multivesicular MHC class II-like compartments (MIIC), release of MHCII(+) vesicles, and expression of MHCII on a dendritic arborization. T-cell-mediated adsorption of MHCII was a saturable process that required close cell proximity, actin polymerization, and a permissive temperature. Adsorbed MHCII existed on vesicles that were intimately associated with the responder cell membrane. T cells bearing adsorbed vesicular MHCII presented antigen and were specifically lysed by CD4(+) T cell responders, but when labeled with anti-MHCII antibody were not susceptible to complement mediated lysis. In summary, this study reveals vesicular compartments associated with synthesis and intercellular exchange of functional MHCII/peptide complexes. PMID- 11902827 TI - Enhancement of antigen driven lymphocyte proliferation secondary to GP41-induced B7 expression on adherent monocytes. AB - HIV-1 viral proteins are known to have immune regulatory effects. The interplay between these molecules and the host immune cells is complex. In this study the immune regulatory effects of gp41 on lymphocyte proliferation were evaluated as a function of the state of the monocyte. It is shown that monocyte adherence to tissue culture plates prevents suppression of lymphocyte proliferation to recall antigen in the presence of gp41. In addition, gp41 can enhance proliferation to low concentrations of Casta antigen when PBL are permitted time to adhere. It is shown that these effects are in part mediated through enhanced expression of the costimulatory molecules B7 and CD40. Cyclosporin A was not able to fully abrogate gp41-enhanced proliferation, indicating participation of a calcium-independent pathway. In addition, concentrations of anti-IL2 receptor antibody sufficient to inhibit maximal proliferation to antigen did not fully inhibit PBL proliferation to antigen that is augmented with gp41. Taken together these results suggest that modification of the monocyte state of activation or differentiation could mediate a response to gp41 that is immune enhancing. PMID- 11902828 TI - Ontogeny of the follicular dendritic cell phenotype and function in the postnatal murine spleen. AB - Follicular dendritic cells (FDCs) represent a unique cell population of antigen trapping cells restricted to follicles within the secondary lymphoid tissues. FDCs appear to be involved in the formation of primary follicles during the ontogeny of lymphoid tissue. We sought to determine the kinetics and tissue distribution of cells in the spleen of newborn mice expressing various differentiation antigens restricted to FDCs using immunohistochemistry with monoclonal antibodies (mAb) against FDCs and in vivo immune complex binding and retention. The earliest FDC-specific marker displayed was the antigenic determinant recognized by the FDC-M1 mAb, which was detectable by Day 3 prior to follicle formation on cells located around the peripheral part of the developing white pulp. The appearance of CD21/35 (complement receptor Type 2 and 1, CR1.2) was observed at the end of the first week, revealing a focal pattern in B-cell rich areas. In addition, at that time there were some FDC-M1-positive cells in the nonfollicular part of the periarteriolar region. The administration of anti horseradish peroxidase antibody followed by soluble antigen HRP into 7-day-old newborn mice resulted in the trapping and retention of immune complexes onto FDCs even in the absence of Fcgamma receptors. The appearance of another FDC-specific marker, FDC-M2, was observed during the second week after birth and was restricted on the cells located in the same area as CR1.2 cells. The Fcgamma receptor Type II appeared on FDCs after the second postnatal week. The above sequence of phenotypic maturation could also be observed in newborns after lethal irradiation at Day 3. This indicates that not only mature FDCs but also their precursors are highly radioresistant, and their phenotypic maturation follows a programmed path that requires only a small number of mature B cells. PMID- 11902830 TI - DNA array and biological characterization of the impact of the maturation status of mouse dendritic cells on their phenotype and antitumor vaccination efficacy. AB - We systematically investigated the impact of the relative maturation levels of dendritic cells (DCs) on their cell surface phenotype, expression of cytokines and chemokines/chemokine receptors (by DNA array and RNase protection analyses), biological activities, and abilities to induce tumor immunity. Mature DCs expressed significantly heightened levels of their antigen-presenting machinery (e.g., CD54, CD80, CD86) and numerous cytokines and chemokines/chemokine receptors (i.e., Flt-3L, G-CSF, IL-1alpha and -1beta, IL-6, IL-12, CCL-2, -3, -4, -5, -17, and -22, MIP-2, and CCR7) and were significantly better at inducing effector T cell responses in vitro. Furthermore, mice vaccinated with tumor peptide-pulsed mature DCs better survived challenge with a weakly immunogenic tumor (8 of 8 survivors) than did mice vaccinated with less mature (3 of 8 survived) or immature (0 of 8 survivors) DCs. Nevertheless, intermediate-maturity DCs expressed substantial levels of Flt-3L, IGF-1, IL-1alpha and -1beta, IL-6, CCL-2, -3, -4, -9/10, -17, and -22, MIP-2, osteopontin, CCR-1, -2, -5, and -7, and CXCR-4. Taken together, our data clearly underscore the critical nature of employing DCs of full maturity for DC-based antitumor vaccination strategies. PMID- 11902829 TI - Expression and regulation of inducible IkappaB kinase (IKK-i) in human fibroblast like synoviocytes. AB - IkappaB kinase (IKK) plays a key role in the regulation of nuclear factor kappaB (NF-kappaB). We previously demonstrated the expression of two kinases, IKK1 and IKK2, in fibroblast-like synoviocytes (FLS) and determined their functional consequences for inflammatory gene expression in vitro and in vivo. Recently, a novel inducible IkappaB kinase has been described, namely, IKK-i or IKK-epsilon, which is functionally and structurally distinct from constitutively expressed IKK1 and IKK2. Therefore, we investigated the expression and regulation of this novel kinase in FLS from patients with rheumatoid arthritis and osteoarthritis. Interestingly, constitutive gene expression and protein expression were observed in all cell lines examined. TNFalpha stimulation for 24 h increased IKK-i expression 7.2 +/- 1.8-fold in FLS (P < 0.02). IL-1 also significantly increased IKK-i gene expression. Time course experiments demonstrated that IKK-i gene expression increased within 3 h of TNFalpha stimulation and persisted for at least 24 h. Dose-response studies showed that as little as 1 ng/ml of TNFalpha increased IKK-i gene expression. Constitutive IKK-1 gene expression was also noted in rheumatoid arthritis, osteoarthritis, and normal synovium. This is the first report demonstrating constitutive expression and cytokine regulation of this novel kinase in primary human synovial cells. PMID- 11902831 TI - Nitric oxide protects thymocytes from gamma-irradiation-induced apoptosis in correlation with inhibition of p53 upregulation and mitochondrial damage. AB - Apoptosis plays a crucial role in clonal deletion in the thymus, and NO has been shown to prevent apoptosis in some cell types. Therefore, we examined the effect of NO on gamma-irradiation-induced thymocyte apoptosis. Treatment of 5 Gy gamma irradiated thymocytes with 1 mM SNAP reduced cell death from 78 to 49% after 8 h incubation (spontaneous cell death in medium control cells was 26%). Coincubation with ZVAD blocked both the spontaneous cell death and the cell death induced by SNAP or gamma-irradiation. The gamma-irradiation-induced increase in caspase 3 and 6 activities was inhibited in the presence of SNAP. The increase in cytosolic cytochrome c as well as the decrease in mitochondrial membrane potential after gamma-irradiation was inhibited in the presence of SNAP. SNAP treatment also decreased the p53 upregulation in gamma-irradiated cells. In summary, we found that NO exerts a protective effect on mouse thymocyte apoptosis induced by gamma irradiation. The mechanism of this protective effect may involve inhibition of p53 upregulation and reduction in mitochondrial damage, with subsequent inhibition of downstream caspase activation. PMID- 11902832 TI - Interleukin-10 contributes development of macrophage suppressor activities by macrophage colony-stimulating factor, but not by granulocyte-macrophage colony stimulating factor. AB - Macrophages are known to possess suppressor activities in immune responses. To determine the effects of GM-CSF and M-CSF on the expression of macrophage suppressor activities, monocyte-derived macrophages cultured with GM-CSF (GM Mphis) were compared with those cultured with M-CSF (M-Mphis) for antigen specific proliferation and interferon-gamma (IFN-gamma) production by lymphocytes. Both GM-Mphis and M-Mphis equally suppressed lymphocyte proliferation, but only M-Mphis suppressed IFN-gamma production in response to purified protein derivative (PPD). M-Mphis, but not GM-Mphis, released IL-10 not only in the course of macrophage differentiation but also in response to PPD after maturation to macrophages. From the results that (i) exogenous IL-10 suppressed IFN-gamma production, but not proliferation of lymphocytes, and that (ii) neutralizing antibody to IL-10 reversed suppressor activities of M-Mphis on IFN-gamma production, but not lymphocyte proliferation, it appeared that IL-10 was the major factor responsible for suppression of IFN-gamma production. Thus, these results suggest that only M-CSF augments IL-10-dependent suppressor activity of macrophages on IFN-gamma production and that both GM-CSF and M-CSF induce IL-10-independent macrophage suppressor activity on lymphocyte proliferation. PMID- 11902833 TI - Peptide affinity for MHC influences the phenotype of CD8(+) T cells primed in vivo. AB - Priming C57BL/6 mice with dominant antigenic peptides of ovalbumin (OVA) or bovine insulin (INS) in complete Freund's adjuvant generates antigen-specific, H 2K(b)-restricted, CD8(+) CTL. OVA-CTL produced type 1 cytokines IFN-gamma and TNF alpha, whereas INS-CTL produced IL-5 and IL-10 with low levels of IL-4 and IFN gamma. Here, we investigate whether differential binding affinities of the OVA and INS peptides to H-2K(b) influence the phenotype of the CD8(+) CTL. OVA(257 264) was found to have significantly higher binding affinity than the INS A chain(12-21) toward K(b). Exchanging the MHC anchor residues between the OVA and INS peptides reversed the K(b) binding capacity of the altered peptides. The lower affinity, altered OVA peptides induced CTL that produced IL-5 and IL-10 in addition to IFN-gamma, whereas high binding affinity, altered INS peptides induced CTL that produced IFN-gamma but not IL-5 or IL-10. These data suggest that MHC binding affinity of peptides can regulate the phenotype of the resulting CD8(+) T cells. PMID- 11902834 TI - Reciprocal "flipping" underlies substrate recognition and catalytic activation by the human 8-oxo-guanine DNA glycosylase. AB - Both 8oxo-guanine and formamidopyrimidines are major products of oxidative DNA damage that can result in the fixation of transversion mutations following replication if left unrepaired. These lesions are targeted by the N-DNA glycosylase hOgg1, which catalyses excision of the aberrant base followed by cleavage of the phosphate backbone directly 5' to the resultant abasic site in a context, dependent manner. We present the crystal structure of native hOgg1 refined to 2.15 A resolution that reveals a number of highly significant conformational changes on association with DNA that are clearly required for substrate recognition and specificity. Changes of this magnitude appear to be unique to hOgg1 and have not been observed in any of the DNA-glycosylase structures analysed to date where both native and DNA-bound forms are available. It has been possible to identify a mechanism whereby the catalytic residue Lys 249 is "primed" for nucleophilic attack of the N-glycosidic bond. PMID- 11902835 TI - Identification of two middle promoters upstream DNA ligase gene 30 of bacteriophage T4. AB - Bacteriophage T4 DNA ligase gene 30 lies in the cluster of prereplicative genes located counterclockwise from map units 149 to 121. Based on the early transcription studies this gene has been considered as a typical early gene of bacteriophage T4. In agreement with this assignment, two strong T4 early promoters, P(E )30.8 (128.6) and P(E )30.7 (128.2), located about 3.1 and 2.7 kb upstream from gene 30 have been revealed by promoter mapping and sequence analysis. In addition, the existence of a putative early promoter just upstream of gene 30 was proposed from the sequence data. However, here we show that the putative early promoter just upstream of gene 30 is, in fact, a T4 middle promoter. Furthermore, we detected one more middle promoter located in the genomic region between early promoter P(E )30.7 (128.2) and DNA ligase gene 30 in the coding region of gene 30.3. Both new middle promoters have differences from the consensus MotA box, while their -10 regions match the sigma(70) consensus sequence very well. The 5' ends of MotA-dependent transcripts directed from these promoters, as well as the kinetics of 5' end accumulation in the cells, have been determined by primer extension analysis. The results of these analyses indicate that both MotA-dependent and MotA-independent promoters control the transcription of T4 DNA ligase gene 30 in vivo. Moreover, we show that the first transcripts for gene 30 are directed from its own middle promoter, P(M)30. PMID- 11902836 TI - Dynalign: an algorithm for finding the secondary structure common to two RNA sequences. AB - With the rapid increase in the size of the genome sequence database, computational analysis of RNA will become increasingly important in revealing structure-function relationships and potential drug targets. RNA secondary structure prediction for a single sequence is 73 % accurate on average for a large database of known secondary structures. This level of accuracy provides a good starting point for determining a secondary structure either by comparative sequence analysis or by the interpretation of experimental studies. Dynalign is a new computer algorithm that improves the accuracy of structure prediction by combining free energy minimization and comparative sequence analysis to find a low free energy structure common to two sequences without requiring any sequence identity. It uses a dynamic programming construct suggested by Sankoff. Dynalign, however, restricts the maximum distance, M, allowed between aligned nucleotides in the two sequences. This makes the calculation tractable because the complexity is simplified to O(M(3)N(3)), where N is the length of the shorter sequence. The accuracy of Dynalign was tested with sets of 13 tRNAs, seven 5 S rRNAs, and two R2 3' UTR sequences. On average, Dynalign predicted 86.1 % of known base-pairs in the tRNAs, as compared to 59.7 % for free energy minimization alone. For the 5 S rRNAs, the average accuracy improves from 47.8 % to 86.4 %. The secondary structure of the R2 3' UTR from Drosophila takahashii is poorly predicted by standard free energy minimization. With Dynalign, however, the structure predicted in tandem with the sequence from Drosophila melanogaster nearly matches the structure determined by comparative sequence analysis. PMID- 11902837 TI - Contribution of the intrinsic curvature to measured DNA persistence length. AB - The persistence length of DNA, a, depends both on the intrinsic curvature of the double helix and on the thermal fluctuations of the angles between adjacent base pairs. We have evaluated two contributions to the value of a by comparing measured values of a for DNA containing a generic sequence and for an "intrinsically straight" DNA. In each 10 bp segment of the intrinsically straight DNA an initial sequence of five bases is repeated in the sequence of the second five bases, so any bends in the first half of the segment are compensated by bends in the opposite direction in the second half. The value of a for the latter DNA depends, to a good approximation, on thermal fluctuations only; there is no intrinsic curvature. The values of a were obtained from measurements of the cyclization efficiency for short DNA fragments, about 200 bp in length. This method determines the persistence length of DNA with exceptional accuracy, due to the very strong dependence of the cyclization efficiency of short fragments on the value of a. We find that the values of a for the two types of DNA fragment are very close and conclude that the contribution of the intrinsic curvature to a is at least 20 times smaller than the contribution of thermal fluctuations. The relationship between this result and the angles between adjacent base-pairs, which specify the intrinsic curvature, is analyzed. PMID- 11902838 TI - X-ray crystallographic studies of serotonin N-acetyltransferase catalysis and inhibition. AB - The structure of serotonin N-acetyltransferase (also known as arylalkylamine N acetyltransferase; AANAT) bound to a potent bisubstrate analog inhibitor has been determined at 2.0 A resolution using a two-edge (Se, Br) multiwavelength anomalous diffraction (MAD) experiment. This acetyl-CoA dependent enzyme is a member of the GCN5-related family of N-acetyltransferases (GNATs), which share four conserved sequence motifs (A-D). In serotonin N-acetyltransferase, motif A adopts an alpha/beta conformation characteristic of the phylogenetically invariant cofactor binding site seen in all previously characterized GNATs. Motif B displays a significantly lower level of conservation among family members, giving rise to a novel alpha/beta structure for the serotonin binding slot. Utilization of a brominated CoA-S-acetyl-tryptamine-bisubstrate analog inhibitor and the MAD method permitted conclusive identification of two radically different conformations for the tryptamine moiety in the catalytic site (cis and trans). A second high-resolution X-ray structure of the enzyme bound to a bisubstrate analog inhibitor, with a longer tether between the acetyl-CoA and tryptamine moieties, demonstrates only the trans conformation. Given a previous proposal that AANAT can catalyze an alkyltransferase reaction in a conformationally altered active site relative to its acetyltransferase activity, it is possible that the two conformations of the bisubstrate analog observed crystallographically correspond to these alternative reaction pathways. Our findings may ultimately lead to the design of analogs with improved AANAT inhibitory properties for in vivo applications. PMID- 11902839 TI - A new algorithm for analysis of oligonucleotide arrays: application to expression profiling in mouse brain regions. AB - Oligonucleotide arrays are a powerful technology for measuring the expression of thousands of genes simultaneously. Improvements in the sensitivity and precision of the measurements, which often pose a challenge to users, would assist the practical application of the technology. Here, we describe a new analysis algorithm for assessing changes in gene expression using oligonucleotide arrays. Changes in expression are detected in terms of the statistical significance (S score) of change, which combines signals detected by multiple probe pairs according to an error model characteristic of oligonucleotide arrays. We show that the S-score is sensitive and reliable, enabling us to obtain more consistent results than with existing methods. Cluster analysis of S-score data of four brain regions exhibits patterns that are more distinctive because of improved data quality. In our case study of two mouse brain regions, over 200 genes were identified to have detectable changes between the ventral tegmental area and the prefrontal cortex. The genes with the most distinctive changes are found to be related to myelin or neurofilament synthesis, calcium signaling, and transcription factors. Many of these findings are in agreement with previous studies, using other techniques, such as in situ hybridization. Overall, our findings suggest that this new algorithm may have broad applicability for improving the analysis of oligonucleotide array data. PMID- 11902840 TI - Crystal structures of artocarpin, a Moraceae lectin with mannose specificity, and its complex with methyl-alpha-D-mannose: implications to the generation of carbohydrate specificity. AB - The seeds of jack fruit (Artocarpus integrifolia) contain two tetrameric lectins, jacalin and artocarpin. Jacalin was the first lectin found to exhibit the beta prism I fold, which is characteristic of the Moraceae plant lectin family. Jacalin contains two polypeptide chains produced by a post-translational proteolysis which has been shown to be crucial for generating its specificity for galactose. Artocarpin is a single chain protein with considerable sequence similarity with jacalin. It, however, exhibits many properties different from those of jacalin. In particular, it is specific to mannose. The structures of two crystal forms, form I and form II, of the native lectin have been determined at 2.4 and 2.5 A resolution, respectively. The structure of the lectin complexed with methyl-alpha-mannose, has also been determined at 2.9 A resolution. The structure is similar to jacalin, although differences exist in details. The crystal structures and detailed modelling studies indicate that the following differences between the carbohydrate binding sites of artocarpin and jacalin are responsible for the difference in the specificities of the two lectins. Firstly, artocarpin does not contain, unlike jacalin, an N terminus generated by post translational proteolysis. Secondly, there is no aromatic residue in the binding site of artocarpin whereas there are four in that of jacalin. A comparison with similar lectins of known structures or sequences, suggests that, in general, stacking interactions with aromatic residues are important for the binding of galactose while such interactions are usually absent in the carbohydrate binding sites of mannose-specific lectins with the beta-prism I fold. PMID- 11902841 TI - Crystal structure of yeast acetohydroxyacid synthase: a target for herbicidal inhibitors. AB - Acetohydroxyacid synthase (AHAS; EC 4.1.3.18) catalyzes the first step in branched-chain amino acid biosynthesis. The enzyme requires thiamin diphosphate and FAD for activity, but the latter is unexpected, because the reaction involves no oxidation or reduction. Due to its presence in plants, AHAS is a target for sulfonylurea and imidazolinone herbicides. Here, the crystal structure to 2.6 A resolution of the catalytic subunit of yeast AHAS is reported. The active site is located at the dimer interface and is near the proposed herbicide-binding site. The conformation of FAD and its position in the active site are defined. The structure of AHAS provides a starting point for the rational design of new herbicides. PMID- 11902842 TI - Base flexibility in HIV-2 TAR RNA mapped by solution (15)N, (13)C NMR relaxation. AB - Binding of the HIV tat protein to the TAR (transactivating response region) RNA element activates transcription of the HIV viral genome. The complex of TAR with argininamide serves as a model for the RNA conformation in the tat-TAR complex. The dynamics of the HIV-2 TAR-argininamide complex was investigated by measurements of the relaxation rates of protonated base carbon and nitrogen nuclei. Six auto-correlation rates as well as cross-correlation rates were measured to map the frequencies of base motion in the HIV-2 TAR-argininamide complex. These measurements reveal a broad range of dynamic heterogeneity exhibited by hexanucleotide loop, the dinucleotide bulge, and the A-form helical regions. U23 in the bulge undergoes the largest dynamic change on binding argininamide, while U25 remains flexible, reflecting the large conformational change that is triggered upon ligand binding. PMID- 11902843 TI - A structural basis for S100 protein specificity derived from comparative analysis of apo and Ca(2+)-calcyclin. AB - Calcyclin is a homodimeric protein belonging to the S100 subfamily of EF-hand Ca(2+)-binding proteins, which function in Ca(2+) signal transduction processes. A refined high-resolution solution structure of Ca(2+)-bound rabbit calcyclin has been determined by heteronuclear solution NMR. In order to understand the Ca(2+) induced structural changes in S100 proteins, in-depth comparative structural analyses were used to compare the apo and Ca(2+)-bound states of calcyclin, the closely related S100B, and the prototypical Ca(2+)-sensor protein calmodulin. Upon Ca(2+) binding, the position and orientation of helix III in the second EF hand is altered, whereas the rest of the protein, including the dimer interface, remains virtually unchanged. This Ca(2+)-induced structural change is much less drastic than the "opening" of the globular EF-hand domains that occurs in classical Ca(2+) sensors, such as calmodulin. Using homology models of calcyclin based on S100B, a binding site in calcyclin has been proposed for the N-terminal domain of annexin XI and the C-terminal domain of the neuronal calcyclin-binding protein. The structural basis for the specificity of S100 proteins is discussed in terms of the variation in sequence of critical contact residues in the common S100 target-binding site. PMID- 11902844 TI - Twist and shear in beta-sheets and beta-ribbons. AB - The structures of the beta-sheets and the beta-ribbons have been analysed using high-resolution protein structure data. Systematic asymmetries measured in both parallel and antiparallel beta-structures include the sheet twist and the strand shear. In order to determine the origin of these asymmetries, numerous interactions and correlations were examined. The strongest correlations are observed for residues in antiparallel beta-sheets and beta-ribbons that form non H-bonded pairs. For these residues, the sheet twist is correlated to the backbone phi angle but not to the psi angle. Our analysis supports the existence of an inter-strand C(alpha)H(alpha)...O weak H-bond, which, together with the CO...HN H bond, constitutes a bifurcated H-bond that links neighbouring beta-strands. Residues of beta-sheets and beta-ribbons in high-resolution protein structures form a distinct region of the Ramachandran plot, which is determined by the formation of the bifurcated H-bond, the formation of an intra-strand O...H(alpha) non-bonded polar interaction, and an intra-strand O...C(beta) steric clash. Using beta-strands parameterised by phi-psi values from the allowed beta-sheet region of the Ramachandran plot, the shear and the right-hand twist can be reproduced in a simple model of the antiparallel and parallel beta-ribbon that models the bifurcated H-bonds specifically. The conformations of interior residues of beta sheets are shown to be subsets of the conformations of residues of beta-ribbons. PMID- 11902845 TI - Protein kinase substrate recognition studied using the recombinant catalytic domain of AMP-activated protein kinase and a model substrate. AB - We have expressed a truncated form of the alpha1 kinase domain of AMP-activated protein kinase (AMPK) in Escherichia coli as a glutathione-S-transferase fusion (GST-KD). A T172D mutant version did not require prior phosphorylation and was utilized for most subsequent studies. We have also created a recombinant substrate (GST-ACC) by expressing 34 residues around the major phosphorylation site (Ser79) on rat acetyl-CoA carboxylase-1/alpha (ACC1) as a GST fusion. This was an excellent substrate that was phosphorylated with similar kinetic parameters to ACC1 by both native AMPK and the bacterially expressed kinase domain. We also constructed a structural model for the binding of the ACC1 sequence to the kinase domain, based on crystal structures for related protein kinases. The model was tested by making a total of 25 mutants of GST-ACC and seven mutants of GST-KD, and measuring kinetic parameters with different combinations. The results reveal that AMPK and ACC1 interact over a much wider region than previously realized (>20 residues). The features of the interaction can be summarised as follows: (i) an amphipathic helix from P-16 to P-5 on the substrate binds in a hydrophobic groove on the large lobe of the kinase; (ii) basic residues at P-6 and P-4 bind to two acidic patches (D215/D216/D217 and E103/D100/E143, respectively), on the large lobe; (iii) a histidine at P+3 interacts with D56 on the small lobe; (iv) the side-chain of P+4 leucine could not be precisely positioned, but a new finding was that asparagine or glutamine could replace a hydrophobic residue at this position. These interactions position the serine residue to be phosphorylated in close proximity to the gamma-phosphate group of ATP. Although based on modelling rather than a determined structure, this represents one of the most detailed studies of the interaction between a kinase and its substrate achieved to date. PMID- 11902846 TI - Integrating the prevention of eating disorders and obesity: feasible or futile? AB - The rate of obesity in adults and youth has doubled in the past 20 years; during this same period there has been an increase in the prevalence of "dysfunctional eating behaviors," including eating disorders and unhealthy weight loss practices. Despite the fact that obesity, eating disorders, and unhealthy weight loss practices are cultivated in the same cultural context-an increasingly "toxic" environment regarding food and weight-these problems are regarded as distinct, with different origins, courses, and approaches to prevention and treatment. In this article, we present conceptual and practical reasons for adopting an integrated approach to the prevention of the spectrum of problems related to eating and weight (i.e., eating disorders, obesity, and unhealthy weight loss practices), suggest personal, socioenvironmental, and behavioral factors to be included in an integrated approach to prevention, and provide some ideas for developing an integrated program using a media literacy/advocacy approach. We conclude with a discussion of challenges to the development of interventions aimed at the broad spectrum of weight-related problems and suggestions for addressing these challenges. PMID- 11902847 TI - Page for patients. Breast feeding. PMID- 11902848 TI - Effects of a tailored health promotion program for female blue-collar workers: health works for women. AB - BACKGROUND: This study assessed the effects of the Health Works for Women (HWW) intervention on improving multiple behaviors including nutrition and physical activity among rural female blue-collar employees in North Carolina. METHODS: Nine small to mid-size workplaces were randomly assigned to either intervention or delayed intervention conditions. After a baseline survey, an intervention consisting of two computer-tailored magazines and a natural helpers program was conducted over 18 months. Delayed worksites received one tailored magazine. Approximately 77 and 76% of baseline respondents completed follow-up surveys at 6 and 18 months, respectively, and 538 women (63%) completed all three surveys. RESULTS: At the 18-month follow-up, the intervention group had increased fruit and vegetable consumption by 0.7 daily servings compared to no change in the delayed group (P < 0.05). Significant differences in fat intake were observed at 6 months (P < 0.05) but not at 18 months. The intervention group also demonstrated improvements in strengthening and flexibility exercise compared to the delayed group. The rates of smoking cessation and cancer screening did not differ between study groups. CONCLUSIONS: The HWW project was a successful model for achieving certain health behavior changes among blue-collar women. PMID- 11902850 TI - A longitudinal study of the effects of tobacco smoking and other modifiable risk factors on ill health in middle-aged and old Americans: results from the Health and Retirement Study and Asset and Health Dynamics among the Oldest Old survey. AB - BACKGROUND: While the effects of smoking and other modifiable risk factors on mortality and specific diseases are well established, their effects on ill health more generally are less known. Using two national, longitudinal surveys, the objective of this study was to analyze the effect of smoking and other modifiable risk factors on ill health, defined in a multidimensional fashion (i.e., disability, impaired mobility, health care utilization, and self-reported health). METHODS: The analyses were based on the Health and Retirement Study (HRS) (12,652 persons 50-60 years old surveyed in 1992, 1994, 1996, and 1998) and the Asset and Health Dynamics among the Oldest Old survey (8,124 persons 60-70 years old surveyed in 1993, 1996, and 1998). RESULTS: Smoking was strongly related to mortality and to ill health, with similar relative effects in the middle-aged and the elderly. There were consistent adverse dose-response relationships between smoking and ill health in the HRS. Persons who had quit smoking at least 15 years prior to the survey were no more likely than never smokers to experience ill health. A dose-response relationship was found between exercise and ill health. For body mass index and alcohol, there were U-shaped relationships with ill health. CONCLUSIONS: Public health efforts designed to encourage smoking cessation should emphasize improvements in ill health in addition to decreased mortality. PMID- 11902851 TI - Antibiotic use and upper respiratory infections: a survey of knowledge, attitudes, and experience in Wisconsin and Minnesota. AB - BACKGROUND: Public attitudes and expectations contribute to inappropriate antibiotic prescribing and antibiotic resistance. This study assessed knowledge, attitudes, and experiences regarding antibiotic use for respiratory infection or illness. METHODS: Random-digit-dialing telephone surveys of adults and parents of children <5 years old were conducted in Wisconsin and Minnesota during 1999. RESULTS: The survey was completed by 405 adults and 275 parents of children <5 years old. The median age was 32 years for parents and 50 years for adults. Seven percent of parents and 17% of adults believed that antibiotics are never or almost never necessary for bronchitis. More than 70% in each group believed that antibiotics are needed for green or yellow nasal drainage, and nearly half of respondents believed that they knew whether an antibiotic was needed before seeing a physician. Exposure to multiple information sources on antibiotic resistance in the past 6 months was independently associated with a knowledge score greater than or equal to the median for nine questions. CONCLUSIONS: The general public has misconceptions regarding indications for antibiotic use, and this may contribute to inappropriate prescribing. Providing multiple and varied antibiotic-related informational messages may increase knowledge of appropriate antibiotic prescribing and decrease patient demand for antibiotics. PMID- 11902852 TI - 5 a day Achievement Badge for African-American Boy Scouts: pilot outcome results. AB - BACKGROUND: Boy Scouts are an important channel to complement school-based programs to enable boys to eat more fruit, 100% juice, and vegetables (FJV) for chronic disease prevention. The "5 a Day Achievement Badge" program was presented on a pilot study basis to African-American Boy Scout troops in Houston. METHODS: Troops were the unit of recruitment and random assignment to treatment and control groups. The badge program was presented in Fall 1997 by trained dietitians and included activities to increase availability and accessibility of fruit and vegetables at scouts' homes, increase preferences for vegetables, and train in the preparation of FaSST (fast, simple, safe, and tasty) recipes. Weekly comic books demonstrated and reinforced what scouts were expected to do at home. A weekly newsletter with recipes was sent to parents. The program was revised and presented to the control group in Winter 1998. Two 24-h recalls were the primary assessment tools. Telephone interviews were conducted with parents. RESULTS: The intervention resulted in a 0.8 FJV serving difference (post values of treatment versus control groups with pre value covaried). CONCLUSIONS: The changes obtained suggest that the intervention was effective in promoting dietary change. PMID- 11902849 TI - Tracking of physical and physiological risk variables among ethnic subgroups from third to eighth grade: the Child and Adolescent Trial for Cardiovascular Health cohort study. AB - BACKGROUND: The Child and Adolescent Trial for Cardiovascular Health (CATCH), a multisite field trial, tested the effectiveness of multiple interventions for cardiovascular disease risk behaviors in children in third through fifth grades. This paper reports the tracking of physiologic variables through eighth grade. METHODS: The cohort began with 5,106 third grade students from diverse ethnic backgrounds: 69% Caucasian, 14% Hispanic, 13% African American, and 4% other. Seventy-two percent of students were remeasured. Measures described are serum lipids, blood pressure, and body anthropometrics. Tracking was examined across three time points (third, fifth, and eighth grades) with a scaled Kendall concordance coefficient and percentage retention within quintiles across time. RESULTS: For the overall sample, tacking was strongest for body mass index (BMI) (Kendall coefficient = 0.86) and weight (0.86), followed by skinfold thicknesses (0.72-0.78), serum lipids (0.67-0.72), and blood pressure (0.45-0.51). For BMI, 96% of students stayed within +/-1 quintile from third to fifth grades; 90% stayed within this range from third to eighth grades. CONCLUSIONS: There were small but noticeable gender and ethnic differences: tracking was stronger among boys and African American students. These results demonstrate that the children's relative level of cardiovascular risk remained stable over a 6-year period. PMID- 11902853 TI - Vitamin C intake and apolipoproteins in a healthy elderly Japanese population. AB - BACKGROUND: Adequate vitamin C (AsA) intake may lower the risk of arteriosclerotic cardiovascular disease, but little is known about its influence on the progression of atherogenic disease in the elderly. METHODS: We examined whether AsA intake was associated with serum lipids, apolipoprotein A-1 (ApoA1) and apolipoprotein B (ApoB), in 680 Japanese elderly persons. RESULTS: There were no significant gender differences among mean serum lipids and apolipoprotein concentrations and intakes of macronutrients. AsA intake had a significant positive association with serum concentrations of high-density cholesterol and ApoA1, but an inverse association with serum concentrations of low-density cholesterol and ApoB, after adjusting for age, body mass index, total energy, and macronutrients. AsA intake was strongly inversely related to ApoA1/ApoB. CONCLUSION: Increased AsA intake could play an important role in lipid composition and could be of potential importance in the genesis and prevention of atherogenic disease in the elderly. PMID- 11902854 TI - Tobacco smoking habits, beliefs, and attitudes among medical students in Tirana, Albania. AB - BACKGROUND: Many surveys throughout the world have evaluated the smoking behaviors, beliefs, and attitudes of medical students, but no information is available from Albania. METHOD: A cross-sectional survey in classroom settings using a self-administered questionnaire was performed at the University of Tirana during October 2000. RESULTS: In the first and fifth years of medical school, 149 (68.5% women) and 185 (55% women) students, respectively, completed the questionnaire, with overall response rates of 82 and 92%. Tobacco smoking prevailed among males. The smoking rates among first-year medical students were 34% among men and 5% among women. Among fifth-year students, 55% of the men and 34% of the women smoked. The percentages of occasional smokers were 29 and 49%, respectively, among the first- and fifth-year students who smoked. Most medical students reported knowing the health hazards of tobacco. Most students in both groups believed that smoking should be restricted in hospitals. Slightly more than half of the students stated that they will regularly advise smokers against smoking in their future jobs. CONCLUSION: Tobacco smoking is common among medical students in Albania. Targeted antismoking training should be mandatory for medical students in Albania. PMID- 11902855 TI - Use of nonphysician health care providers for skin cancer screening in the primary care setting. AB - BACKGROUND: Nonphysician health care providers are in an optimal position to provide cancer prevention and screening services. METHODS: We conducted a survey of primary care physicians to determine physician use and amenability to use of nonphysician health care providers to perform skin cancer screening in comparison with other cancer screening examinations. RESULTS: A total of 1,363 eligible physicians completed the survey. Of these, 631 physicians (46%) reported a nurse practitioner or physician assistant performing at least one type of cancer screening examination on their patients. Twenty-nine and 22% of all physicians reported nurse practitioners or physician assistants performing skin cancer screening, respectively. Family physicians were more likely to use nurse practitioners and physician assistants to perform these cancer screening examinations than internists (chi(2) test, P = 0.001 for each examination). Skin examinations were performed less frequently by nurse practitioners and physician assistants than all other cancer screening examinations. A total of 73-79% of family physicians and 60-70% of internists were amenable to having a nonphysician health care provider perform one or more of these examinations. CONCLUSIONS: Primary care physicians are currently utilizing nonphysician health care providers to perform cancer screening examinations and the majority of those surveyed are amenable to the use of these providers for such examinations. This suggests that one possible strategy for increasing skin cancer screening is through an expanded role of nonphysician health care providers. PMID- 11902856 TI - Men's and women's knowledge and perceptions of breast cancer and mammography screening. AB - BACKGROUND: Although most men are not directly affected by breast cancer, they participate in decisions influencing breast cancer screening and contribute to shaping the social norm on mammography screening. This study tested the hypothesis that men may be less knowledgeable than women about breast cancer and mammography and have less favorable perceptions of mammography screening. METHODS: A survey was mailed to 952 women and 370 men aged 40 to 80 years, randomly selected from the general population of Geneva, Switzerland. Information collected included knowledge and perceptions about breast cancer and mammography, familiarity with screening recommendations, and perceived usefulness of an organized screening program. RESULTS: Men were almost as knowledgeable as women about breast cancer (difference in z-scores: -0.12; 95% CI: -0.25, 0.02; P = 0.10) and breast cancer screening (difference: -0.12; 95% CI: -0.25, 0.02; P = 0.09). Both men and women perceived mammography screening to be useful, but men had significantly higher z-scores of positive attitude toward mammography than women (difference: 0.28; 95% CI: 0.14, 0.42; P < 0.001). Younger and more educated respondents of either sex were both more knowledgeable and more favorably inclined toward screening. CONCLUSION: Men were as knowledgeable about breast cancer and mammography screening as women but had more favorable attitudes toward breast cancer screening than women. Actions to strengthen community support for mammography screening programs should primarily target older and less educated persons of either sex. PMID- 11902858 TI - Smoking behaviors and regular source of health care among African Americans. AB - OBJECTIVES: The purpose of the study was to determine associations between having a regular source of health care, advice from a physician to quit smoking, and smoking-related behaviors among African American smokers. METHODS: A secondary analysis was conducted on data obtained from an intervention study with a posttest assessment of the effectiveness of smoking status as a vital sign. The setting was an adult walk-in clinic at a large inner-city hospital and 879 African American adult current smokers were examined. RESULTS: Among African American smokers, there was an association between having a regular source of health care and planning to quit smoking within the next 30 days (OR = 1.46; 95% CI: 1.04-2.05), receiving physician advice to quit (OR = 1.46; 95% CI: 1.02 2.10), and smoking < or =10 cigarettes a day (OR = 1.42; 95% CI: 1.00-2.03). CONCLUSIONS: African American current smokers with a regular source of health care were further along the quitting process than those without a regular source of health care. Our findings indicate a potential benefit of complementing programs that increase physician cessation advice rates with policies that increase rates of health insurance and the likelihood that individuals have a regular source of health care. PMID- 11902859 TI - Water-stabilized cavitands. AB - Tetrabenzimidazole cavitands 4 were prepared by condensation of ortho esters with octaamino cavitand 3 in 70-80% yield. Molecular modeling predicted that no intramolecular hydrogen bonds are possible between the imidazole fragments in the vase conformation of 4. Instead, this conformation provides four perfect binding sites for hydroxyl-containing molecules through an N-H---O-H---N pattern. Such interactions provide the means for sealing the cavitand's cavity. Accordingly, dry compounds 4 are not soluble in dry CDCl3 but readily dissolve upon addition of small amounts of alcohols or by saturation of the solution with water. 1H NMR spectroscopy revealed that in these solutions molecules 4 adopt a vase conformation while 1D GOESY experiments revealed their monomeric nature. In water saturated CDCl3, these cavitands 4 form kinetically stable 1:1 inclusion complexes with tetramethylphosphonium bromide and triethylammonium chloride in which the cation is incorporated into the pi-basic cavity. Thus, cavitands 4 are a novel class of open-ended molecular containers capable of the formation of highly kinetically stable complexes upon assistance by hydrogen-bonding water molecules. PMID- 11902857 TI - Use of mammographic screening by HIV-infected women in the Women's Interagency HIV Study (WIHS). AB - BACKGROUND: Although HIV-positive women may be less likely than women in general to receive mammography due to socioeconomic disadvantage, HIV diagnosis may increase opportunities for medical interactions which encourage mammography. METHODS: HIV-positive (2,059) and HIV-negative (569) Women's Interagency HIV Study (WIHS) participants reported ever/never history of mammography at baseline (in 1994, 1995) and, at each 6-month follow-up visit, if they had been screened since their last visit. National Health Interview Survey (NHIS) data for 1994 were used to compare WIHS participants to U.S. women. Factors independently related to mammography were determined using logistic regression for baseline data and proportional hazards for follow-up data. Results were adjusted for age. RESULTS: Among women > or =40, fewer WIHS women, regardless of HIV status, reported screening than U.S. women (67% HIV-positive, 62% HIV-negative, 79% NHIS; P < 0.0001). First-time screening while on study was associated with being HIV positive [rate ratio (95% confidence interval) = 1.6 (1.1, 2.3)]. Factors independently associated with screening were related to health care access and usage. CONCLUSIONS: WIHS women, a disadvantaged population, reported less mammography than the general population. HIV-positive women reported more screening than HIV-negative women, possibly because of greater opportunity to interact with the health care system. PMID- 11902860 TI - DNA-based magnetic nanoparticle assembly acts as a magnetic relaxation nanoswitch allowing screening of DNA-cleaving agents. AB - Monodisperse magnetic nanoparticles conjugated with complementary oligonucleotide sequences self-assemble into stable magnetic nanoassemblies resulting in a decrease of the spin-spin relaxation times (T2) of neighboring water protons. When these nanoassemblies are treated with a DNA cleaving agent, the nanoparticles become dispersed, switching the T2 of the solution back to original values. These qualities render the developed nanoparticles and their nanoassemblies as magnetic relaxation switches capable of screening for DNA cleaving compounds by magnetic resonance methods such as MRI and NMR. PMID- 11902861 TI - Use of chirally modified zeolites and crystals in photochemical asymmetric synthesis. AB - Three different approaches to asymmetric induction in the cis-to-trans photoisomerization of a number of 1-benzoyl-2,3-diphenylcyclopropane derivatives are reported: the use of chiral inductors and covalent chiral auxiliaries in MY zeolites and the use of ionic chiral auxiliaries in crystals. High levels of asymmetric induction were achieved using the latter two methods-up to 71% through the use of covalent chiral auxiliaries in zeolites and a remarkable 99% via the solid state ionic chiral auxiliary approach. In the zeolite method, the diastereomeric excess was found to depend strongly on the nature of the zeolite cation, M(+), and in the ionic chiral auxiliary approach, evidence is presented that it is the fixed orientation of the benzoyl group with respect to the cyclopropane ring that controls enantioselectivity in the crystalline state-a finding that is directly relevant to theoretical work on this topic. PMID- 11902862 TI - Direct H atom abstraction from spore photoproduct C-6 initiates DNA repair in the reaction catalyzed by spore photoproduct lyase: evidence for a reversibly generated adenosyl radical intermediate. AB - Spore photoproduct (SP) lyase, which catalyzes the direct reversal of SP (5 thyminyl-5,6-dihydrothymine) to thymine monomers, is the only identified nonphotoactivatable pyrimidine dimer lyase. Unlike DNA photolyase, SP lyase does not contain a flavin cofactor and does not require light for activation. Instead, preliminary studies point to the presence of an iron-sulfur cluster in SP lyase and the requirement for S-adenosylmethionine (AdoMet) for catalytic activity, suggesting that SP lyase belongs to the growing group of iron-sulfur cluster and AdoMet-dependent radical enzymes. Here we provide evidence for the role of AdoMet as a reversible deoxyadenosyl radical generator, which initiates repair by hydrogen atom abstraction from C-6 of SP. Reaction of 6-(3)H-SP, but not methyl (3)H-SP, with SP lyase and AdoMet results in transfer of (3)H to AdoMet, while no tritiated 5'-deoxyadenosine is observed. When 5'-tritiated AdoMet is used in the reaction with unlabeled SP, transfer of (3)H into the repaired thymine monomers is observed. These results point to the reversible generation of a 5' deoxyadenosyl radical intermediate, which reacts directly with the DNA lesion to initiate a radical-mediated beta-scission. We also demonstrate that AdoMet is a catalytic cofactor that is not consumed during turnover. Together, these results support a novel radical-based mechanism for the repair of UV-induced DNA damage. PMID- 11902863 TI - Receptor-bound conformation of an alpha(5)beta(1) integrin antagonist by (15)N edited 2D transferred nuclear overhauser effects. AB - We report the results of (15)N-edited 2D transferred NOE experiments of the partially (15)N-labeled alpha(5)beta(1) antagonist c[Mpa(15)N-Arg-(15)N-Gly-(15)N Asp-(15)N-Asp-(15)N-Val-Cys]-NH(2) (Mpa denotes mercaptopropionic acid) in the presence of the native alpha(5)beta(1) receptor. The alpha(5)beta(1) integrin receptor is believed to be involved in tumor metastasis and the rational design of alpha(5)beta(1) integrin antagonist is therefore of considerable interest. Our experiments provide insight into the alpha(5)beta(1) receptor-bound conformation of the antagonist c[MpaRGDDVC]-NH2 and will be important for the design of novel antagonists. PMID- 11902864 TI - Changing DNA grooves--a 1,4,5,8-naphthalene tetracarboxylic diimide bis intercalator with the linker (beta-Ala)(3)-Lys in the minor groove. AB - We have been investigating a modular, threading DNA polyintercalator design based upon the 1,4,5,8-naphthalene tetracarboxylic diimide (NDI) intercalating unit. Previously, we have reported the NMR analysis of a bis-intercalator-DNA complex in which the peptide linker between NDI units was found to occupy the DNA major groove (Guelev, Lee, Sorey, Hoffman, Iverson, Chem. Biol. 2001, 8, 415-425). Here we describe the NMR analysis of a complex between a related bis-intercalator known to display altered DNA sequence specificity. In this case, the linker resides in the DNA minor groove. We have thus shown that within this set of sequence specific bis-intercalators, both DNA grooves can be accessed, setting the stage for longer threading polyintercalators designed to have linkers occupying both grooves in an alternating fashion. PMID- 11902865 TI - Chi(1) rotamer populations and angles of mobile surface side chains are accurately predicted by a torsion angle database potential of mean force. AB - The equilibrium angles and distributions of chi(1) rotamers for mobile surface side chains of the small, 63-residue, B1 domain of protein L have been calculated from the static crystal structure by rigid body/torsion angle simulated annealing using a torsion angle database potential of mean force and compared to those deduced by Monte Carlo analysis of side chain residual dipolar couplings measured in solution. Good agreement between theory and experiment is observed, indicating that for side chains undergoing rotamer averaging that is fast on the chemical shift time scale, the equilibrium angles and distribution of chi(1) rotamers are largely determined by the backbone phi/psi torsion angles. PMID- 11902866 TI - Enantioselective synthesis of medium-ring heterocycles, tertiary ethers, and tertiary alcohols by Mo-catalyzed ring-closing metathesis. AB - The Mo-catalyzed asymmetric ring-closing metathesis (ARCM) of various achiral trienes leads to the formation of medium-ring unsaturated heterocycles in high yield and with excellent enantioselectivity. Reactions may be carried out on gram scale and in the absence of solvent. The unsaturated siloxanes obtained enantioselectively can be readily functionalized to obtain synthetically useful and difficult-to-access tertiary alcohols. PMID- 11902867 TI - Dynamic kinetic resolution via dual-function catalysis of modified cinchona alkaloids: asymmetric synthesis of alpha-hydroxy carboxylic acids. AB - A highly enantioselective catalytic transformation of racemic alpha-hydroxy acids to optically active alpha-hydroxy acids is reported. A new procedure was developed for the condensation of racemic alpha-hydroxy acids with trichloromethyl chloroformate (diphosgene) at room temperature in the presence of activated charcoal to form 5-substituted-1,3-dioxolane-2,4-diones in 90-100% yield. An efficient dynamic kinetic resolution of 5-aryl dioxolanediones was realized via a modified cinchona alkaloid-catalyzed alcoholytic opening of the dioxolanedione ring, generating a variety of optically active alpha-hydroxy esters in 91-96% ee and 61-85% chemical yield. In this dynamic kinetic resolution, the modified cinchona alkaloid was found to serve dual catalytic roles, mediating both the rapid racemization of the 5-aryl dioxolanediones and the enantioselective alcoholytic ring opening of the 5-aryl dioxolanediones. Consequently, both enantiomers of the 5-aryl dioxolanediones were converted to highly enantiomerically enriched aromatic alpha-hydroxy esters in yields (61 85%), far exceeding the maximum of 50% for a normal kinetic resolution. This development not only represents an expansion of the scope of asymmetric acyl transfer catalysis of synthetic catalysts but also provides a new approach for the development of efficient chemical dynamic kinetic resolutions promoted by a single catalyst. 5-Alkyl dioxolanediones were resolved by a conventional but highly enantioselective kinetic resolution to provide alpha-hydroxy acids and esters in high optical purity and good yields. PMID- 11902868 TI - The first fullerene-metal sandwich complex: an unusually strong electronic communication between two C(60) cages. AB - Reaction of Rh(6)(CO)(9)(dppm)(2)(mu(3)-eta(2),eta(2),eta(2)-C(60)) (1) with C(60) in refluxing chlorobenzene followed by treatment with CNR (R = CH(2)C(6)H(5)) at room temperature affords the first fullerene-metal sandwich complex Rh(6)(CO)(5)(dppm)(2)(CNR)(mu(3)-eta(2),eta(2),eta(2)-C(60))(2) (2). Compound 2 has been characterized by an X-ray diffraction study. Electrochemical study of 2 reveals six well-separated reversible redox couples localized at C(60) cages due to a strong electronic communication between the two C(60) centers via the Rh(6) cluster spacer. PMID- 11902869 TI - Organotellurium compounds as novel initiators for controlled/living radical polymerizations. Synthesis of functionalized polystyrenes and end-group modifications. AB - Polymer-end mimetic organotellurium compounds initiate controlled/living radical polymerization of styrene derivatives that allows accurate molecular weight control with defined end-groups. Transformations of the end-groups via radical and ionic reactions provide a variety of end-group modified polystyrenes. PMID- 11902870 TI - Three-component cycloadditions: the first transition metal-catalyzed [5+2+1] cycloaddition reactions. AB - Prompted by our studies of transition metal-catalyzed [4+4], [4+2], [5+2], and [6+2] cycloadditions and by the view that these two-component reactions could be intercepted by a third component of one or more atoms, a new three-component transition metal-catalyzed cycloaddition is described. This new [5+2+1] cycloaddition proceeds in good to excellent yield and with high or complete regioselectivity with a variety of carbonyl-substituted alkynes to give bicyclo[3.3.0]octenone adducts, resulting from transannular closure of the intermediate eight-membered-ring cycloadduct. Effects of concentration, temperature, pressure, and catalyst loading on the efficiency of the reaction are discussed. This process provides access to complex building blocks for synthesis based on simple, readily available components. PMID- 11902872 TI - Selected-control hydrothermal synthesis of alpha- and beta-MnO(2) single crystal nanowires. AB - A selective-control hydrothermal method has been developed in the preparation of alpha- and beta-MnO(2) single-crystal nanowires. The crystal structure and morphology of the final products can be influenced by the concentration of NH(4)(+) and SO(4)(2-). PMID- 11902871 TI - A pseudo-Jahn-Teller distortion in an Mo(2)(mu(2)-O)(2) ring having the shortest Mo(IV)-Mo(IV) double bond. AB - Four structures of edge-sharing bioctahedral compounds of the type Mo(2)(mu(2) DArF)(2)(eta(2)-L-L)(2)(mu(2)-O)(2), where DArF is an anion of an N,N' diarylformamidine and L-L is a chelating acetate or DArF group, are reported here. The cores of the ring formed by the Mo(2)(mu(2)-O)(2) are very similar with very short Mo-Mo distances of 2.306[2] A. These are consistent with the presence of a Mo=Mo double bond of the type sigma(2)pi(2). As expected for these electronic configurations, the compounds are diamagnetic. The most striking characteristic is the distortion of the Mo(2)(mu(2)-O)(2) ring where a set of two Mo-O distances are significantly shorter then the other set (by ca. 0.05 A). This D(2)(h)--> C(2)(h) distortion is explained on the basis of a pseudo-Jahn-Teller effect. PMID- 11902873 TI - Asymmetric catalysis of hetero-ene reactions with tridentate Schiff base chromium(III) complexes. AB - Tridentate Schiff base chromium(III) complex 1 catalyzes the asymmetric hetero ene reaction between aryl aldehydes and either 2-methoxypropene or 2 trimethylsilyloxypropene to provide a series of beta-hydroxyenol ether products in high yields and enantioselectivities. X-ray crystallographic analysis of a closely related chromium complex reveals a bridged, dimeric structure bearing aquo-bound six-coordinate Cr(III) centers. A mechanism is proposed wherein water dissociation is effected by means of a chemical desiccant (BaO or silyl enol ether), thereby revealing the site for aldehyde complexation. PMID- 11902874 TI - Polymer mediated self-assembly of magnetic nanoparticles. AB - We present a simple polymer-mediated process of assembling magnetic FePt nanoparticles on a solid substrate. Alternatively absorbing the PEI molecule and FePt nanoparticles on a HO-terminated solid surface leads to a smooth FePt nanoparticle assembly with controlled assembly thickness and dimension. Magnetic measurements show that the thermally annealed FePt nanoparticle assembly as thin as three nanoparticle layers is ferromagnetic. The magnetization direction of this thin FePt nanoparticle assembly is readily controlled with the laser assisted magnetic writing. The reported process can be applied to various substrates, nanoparticles, and functional macromolecules and will be useful for future magnetic nanodevice fabrication. PMID- 11902875 TI - Effect of molecule-metal electronic coupling on through-bond hole tunneling across metal-organic monolayer-semiconductor junctions. AB - Using Hg/alkyl-chain-monolayer/p-Si devices we find that the type of contact between the chains and the electrodes (chemical bonding or not) is of critical importance for electronic transport across the junctions. As the semiconductor is p-type, the transport is that of holes. In agreement with theory we find that holes tunnel more efficiently through alkyl chains than do electrons. PMID- 11902876 TI - Enantioselective addition of ketene silyl acetals to nitrones catalyzed by chiral titanium complexes. Synthesis of optically active beta-amino acids. AB - A chiral titanium complex, Ti(O-i-Pr)(4)/BINOL/tert-butylcatechol, catalyzes enantioselective addition reaction of ketene silyl acetals to nitrones to give optically active beta-amino acid derivatives which are biologically active compounds and useful synthetic intermediates of natural products and pharmaceuticals such as beta-lactam antibiotics. The combined process of catalytic oxidation of secondary amines and enantioselective carbon-carbon bond formation of nitrones thus obtained with ketene silyl acetals provides a useful two-step method for the synthesis of optically active beta-amino acid derivatives and related nitrogen compounds. PMID- 11902877 TI - Interactions of aziridines with nickel complexes: oxidative-addition and reductive-elimination reactions that break and make C-N bonds. AB - Reaction of the N-tosylaziridines (p-CH(3)C(6)H(4)SO(2))NCH(2)CHR (1a, R = H; 1b, R = Me; 1c, R = n-Bu; 1d, R = i-Pr) with (bpy)Ni(cod) (2; bpy = 2,2'-bipyridine; cod = 1,5-cyclooctadiene) or (bpy)NiEt(2) (3) results in elimination of cod or butane from 2 and 3, respectively, and oxidative addition of an aziridine C-N bond to give the azametallacyclobutane complexes (bpy)Ni(NTosCHRCH(2)) (4a, R = H; 4b, R = Me; 4c, R = n-Bu; 4d, R = i-Pr) as maroon solids in 50-70% isolated yields. The structure of 4b exhibits a puckered four-membered azametallacycle containing a pyramidal nitrogen and with Ni-N(1) = 1.911(5) A; the tosyl group on N and the methyl substituent on the adjacent C are disposed in an anti conformation. The monodeuterated aziridine syn-(p-CH(3)C(6)H(4)SO(2))NCHDCH-n-Bu (1e) reacts with either 2 or 3 to give (bpy)Ni[NTosCH(n-Bu)CHD] (4e) in 60-65% yield, having an anti arrangement of the methine and methylene protons in the azametallacycle, and indicates that >95% inversion of stereochemistry has occurred at the methylene carbon during the oxidative-addition reaction. When the azametallacyclobutane complexes 4a-e are exposed to oxygen, oxidatively induced reductive elimination ensues, giving the free aziridines in 30-60% isolated yields. In the oxidation of 4e, the product aziridine is spectroscopically identical to its parent, 1e, indicating the elimination that forms the C-N bond also proceeds with inversion of stereochemistry (approximately 92% by (1)H NMR) at the methylene carbon. PMID- 11902878 TI - Dynamic kinetic resolution via asymmetric conjugate reduction: enantio- and diastereoselective synthesis of 2,4-dialkyl cyclopentanones. AB - Herein, we report the kinetic and the dynamic kinetic resolutions of racemic 3,5 dialkyl-2-cyclopenten-1-ones. Kinetic resolution, with good selectivity factors (25-52), was achieved by conjugate reduction with catalytic CuCl/NaOt-Bu/(S)-p tol-BINAP and stoichiometric quantities of poly(methylhydrosiloxane) (PMHS). When stoichiometric amounts of NaOt-Bu and t-BuOH were included in the reaction mixture, rapid racemization of the starting material occurred allowing for the dynamic kinetic resolution of the cyclopentenone substrates. In this process, chiral 2,4-dialkylcyclopentanones were isolated with high stereoselectivity (ee > or = 91%, dr > or = 90:10) and in high yield (> or =89%). PMID- 11902879 TI - Nonactin biosynthesis: the initial committed step is the condensation of acetate (malonate) and succinate. AB - Nonactin is a macrotetrolide antibiotic produced by Streptomyces griseus subsp. griseus ETH A7796 that has shown activity against the P170-glycoprotein efflux pump associated with multiple drug resistant cancer cells. Nonactin is a polyketide, albeit a highly atypical one. The structure is composed of two units of each of the enantiomers of nonactic acid, arranged in a macrocycle, so that the molecule has S4 symmetry and is achiral. The monomer units, (+)- and (-) nonactic acid, are derived from acetate, succinate, and propionate, although the exact details of the assembly process are quite unclear. We have used feeding experiments with a series of multiple stable isotope labeled precursors to elucidate the details of the first committed step of nonactic acid biosynthesis. We have found that the (13)C label from 3-ketoadipate is incorporated specifically into both nonactic acid and its homologue, homononactic acid. The data conclusively show that the first committed step of nonactin biosynthesis is the coupling of a succinate derivative with either acetate or malonate. The differentiation into either nonactate or homononactate occurs after the initial condensation; the homologues are not derived from use of a different "starter unit" by the nonactate polyketide synthase. The first step of nonactin biosynthesis involves achiral intermediates; differentiation between the known enantiocomplementary biosynthesis pathways to form each enantiomer of the precursor monomer units likely occurs after the initial condensation reaction. PMID- 11902881 TI - Measurement of electrostatic interactions in protein folding with the use of protein charge ladders. AB - This paper describes a new method for the measurement of the role of interactions between charged groups on the energetics of protein folding. This method uses capillary electrophoresis (CE) and protein charge ladders (mixtures of protein derivatives that differ incrementally in number of charged groups) to measure, in a single set of electrophoresis experiments, the free energy of unfolding (DeltaG(D-N)) of alpha-lactalbumin (alpha-LA) as a function of net charge. These same data also yield the hydrodynamic radius, R(H), and net charge measured by CE, Z(CE), of the folded and denatured proteins. Alpha-LA unfolds to a compact denatured state under mildly alkaline conditions; a small increase in R(H) (11%, 2 A) coincides with a large increase in Z(CE) (71%, -4 charge units), relative to the folded state. The increase in Z(CE), in turn, predicts a large pH dependence of free energy of unfolding (-22 kJ/mol per unit increase in pH), due to differences in proton binding in the folded and denatured states. The free energy of unfolding correlates with the square of net charge of the members of the charge ladder. The differential dependence of DeltaG(D-N) on net charge for holo alpha-LA, (partial differential) DeltaG(D-N)/(partial differential)Z = -0.14Z kJ/mol per unit of charge. This dependence of DeltaG(D-N) on net charge is a result of a net electrostatic repulsion among charge groups on the protein. These results, together with data from pH titrations, show that both the effects of electrostatic repulsion and differences in proton binding in the folded and denatured states can play an important role in the pH dependence of this protein; the relative magnitude of these effects varies with pH. The combination of charge ladders and CE is a rapid and efficient tool that measures the contributions of electrostatics to the energetics of protein folding, and the size and charge of proteins as they unfold. All this information is obtained from a single set of electrophoresis experiments. PMID- 11902880 TI - A noncovalent approach to antiparallel beta-sheet formation. AB - Four tripeptide chains, when attached to the same end of a hydrogen-bonded duplex (1.2) with the unsymmetrical, complementary sequences of ADAA/DADD, have been brought into proximity, leading to the formation of four hybrid duplexes, 1a.2a, 1a.2b, 1b.2a, and 1b.2b, each of which contains a two-stranded beta-sheet segment. The extended conformations of the peptide chains were confirmed by 1D and 2D NMR. The peptide strands stay registered through hydrogen bonding and the beta-sheets are stabilized by side chain interactions. Two-dimensional NMR data also indicate that the duplex template prevents further aggregation in the peptide segment. When the peptide chains are attached to the two different termini of the duplex template, NMR studies show the presence of a mixture with no clearly defined conformations. In the absence of the duplex template, the tripeptides are found to associate randomly. Finally, isothermal titration calorimetry studies revealed that the hybrid duplex 1a.2a was more stable than either the duplex template or the peptides alone. PMID- 11902882 TI - Influence of silaproline on peptide conformation and bioactivity. AB - The analogue gamma-(dimethylsila)-proline, denoted silaproline (Sip), was synthesized in both enantiomerically pure forms by diastereoselective alkylation of a chiral glycine equivalent with use of Schollkopf's bis-lactim ether method. The effect of replacing a proline residue in model peptides by this new proline surrogate has been examined in the crystal state by X-ray diffraction and in solution by IR absorption and NMR techniques. Silaproline and proline-containing sequences exhibit very similar conformational properties. Silaproline was also substituted for proline in a neurotensin (8-13) analogue that retained biological activity and exhibited enhanced resistance to biodegradation. PMID- 11902883 TI - Acyclic diastereoselection in prochiral radical addition to prochiral olefins. AB - The stereochemical preference (syn or anti) when prochiral radicals add to prochiral acceptors is of fundamental interest. The primary focus of this research was to determine which factors influence the relative stereochemistry between the beta and gamma chiral centers when these are formed concurrently. While moderate diastereoselectivity was found for addition of alkyl (6a-d) and alpha-alkoxy radicals (16a-c) (< or =6:1 syn) to acceptors 4, 7, 8, 10, and 14, consistently high selectivity was observed with less reactive halogenated radicals (6f,g) (>15:1 anti). Steric influence in alkyl radical additions was difficult to evaluate due to decreased reactivity when using bulky reaction partners; however, more reactive alpha-alkoxy radicals, it was found that increasing steric bulk leads to moderate increases in selectivity. In addition, higher selectivity was observed when employing lanthanide Lewis acids whose environment (reactivity) was modified using achiral additives, suggesting a potentially simple means for selectivity enhancements in radical reactions. Overall these results indicate that significant stereoelectronic effects are necessary to achieve high levels of selectivity in prochiral radical additions to prochiral acceptors. PMID- 11902884 TI - Using equilibrium isotope effects to detect intramolecular OH/OH hydrogen bonds: structural and solvent effects. AB - A comparative (1)H NMR study of partially deuterated 1,3- and 1,4-diols has demonstrated that intramolecular hydrogen bonds of different geometry can give rise to equilibrium isotope shifts of opposite sign in hydrogen-bond-accepting solvents such as DMSO-d(6), acetone-d(6), and THF-d(8). The sign inversion is interpreted in terms of the ability of solvent molecules to form competitive intermolecular hydrogen bonds with the diol and in terms of the limiting chemical shifts for the interior and exterior hydroxyl groups. Deuterium is shown to prefer the intermolecular solvent hydrogen bond by 10.9 +/- 0.5 cal/mol for 1,4 diol 3 dissolved in DMSO-d(6) at room temperature. Pyridine-d(5) is shown to be capable of amplifying positive (downfield) isotope shifts measured in DMSO-d(6), in some cases by as much as a factor of 3. Its use is demonstrated for the assignment of the syn or anti relative configuration of 2,4-pentanediol and for the amplification of isotope shifts used to detect intramolecular hydrogen bonds in alpha- and beta-cyclodextrin. Studies in apolar solvents such as CD(2)Cl(2) and benzene-d(6) reveal that the isotope shift is negative (upfield) for all hydrogen bond geometries studied. Larger isotope shifts are measured in benzene d(6), and a rationale for this amplification is presented. The use of apolar solvents is particularly useful for assigning the syn or anti configuration of 2,4-pentanediol. PMID- 11902885 TI - Switching "on" and "off" the expression of chirality in peptide rotaxanes. AB - The hydrogen-bond-directed synthesis, X-ray crystal structures, and optical properties of the first chiral peptide rotaxanes are reported. Collectively these systems provide the first examples of single molecular species where the expression of chirality in the form of a circular dichroism (CD) response can selectively be switched "on" or "off", and its magnitude altered, through controlling the interactions between mechanically interlocked submolecular components. The switching is achievable both thermally and through changes in the nature of the environment. Peptido[2]rotaxanes consisting of an intrinsically achiral benzylic amide macrocycle locked onto various chiral dipeptide (Gly-L Ala, Gly-L-Leu, Gly-L-Met, Gly-L-Phe, and Gly-L-Pro) threads exhibit strong (10 20k deg cm(2) dmol(-1)) negative induced CD (theta;) values in nonpolar solvents (e.g. CHCl(3)), where the intramolecular hydrogen bonding between thread and macrocycle is maximized. In polar solvents (e.g., MeOH), where the intercomponent hydrogen bonding is weakened, or switched off completely, the elliptical polarization falls close to zero in some cases and can even be switched to large positive values in others. Importantly, the mechanism of generating the switchable CD response in the chiral peptide rotaxanes is also determined: a combination of semiempirical calculations and geometrical modeling using the continuous chirality measure (CCM) shows that the chirality is transmitted from the amino acid asymmetric center on the thread via the macrocycle to the C terminal stopper of the rotaxane. This understanding could have important implications for other areas where chiral transmission from one chemical entity to another underpins a physical or chemical response, such as the seeding of supertwisted nematic liquid crystalline phases or asymmetric synthesis. PMID- 11902886 TI - Total synthesis of (-)-tetrazomine. Determination of the stereochemistry of tetrazomine and the synthesis and biological activity of tetrazomine analogues. AB - The first total synthesis of the potent antitumor antibiotic (-)-tetrazomine has been accomplished. A new method for the formation of the allylic amine precursor to an azomethine ylide has been developed and exploited in an efficient [1,3] dipolar cycloaddition to afford the key tetracyclic intermediate used in the synthesis of (-)-tetrazomine. Several analogues of tetrazomine have been synthesized and tested for antimicrobial and biochemical activity. PMID- 11902887 TI - Formation and stability of enolates of acetamide and acetate anion: an Eigen plot for proton transfer at alpha-carbonyl carbon. AB - Second-order rate constants were determined in D(2)O for deprotonation of acetamide, N,N-dimethylacetamide, and acetate anion by deuterioxide ion and for deprotonation of acetamide by quinuclidine. The values of k(B) = 4.8 x 10(-8) M( 1) s(-1) for deprotonation of acetamide by quinuclidine (pK(BH) = 11.5) and k(BH) = 2-5 x 10(9) M(-1) s(-1) for the encounter-limited reverse protonation of the enolate by protonated quinuclidine give pK(a)(C) = 28.4 for ionization of acetamide as a carbon acid. The limiting value of k(HOH) = 1 x 10(11) s(-1) for protonation of the enolate of acetate anion by solvent water and k(HO) = 3.5 x 10(-9) M(-1) s(-1) for deprotonation of acetate anion by HO(-) give pK(a)(C) approximately 33.5 for acetate anion. The change in the rate-limiting step from chemical proton transfer to solvent reorganization results in a downward break in the slope of the plot of log k(HO) against carbon acid pK(a) for deprotonation of a wide range of neutral alpha-carbonyl carbon acids by hydroxide ion, from -0.40 to -1.0. Good estimates are reported for the stabilization of the carbonyl group relative to the enol tautomer by electron donation from alpha-SEt, alpha-OMe, alpha-NH(2), and alpha-O(-) substituents. The alpha-NH(2) and alpha-OMe groups show similar stabilizing interactions with the carbonyl group, while the interaction of alpha-O(-) is only 3.4 kcal/mol more stabilizing than for alpha OH. We propose that destabilization of the enolate intermediates of enzymatic reactions results in an increasing recruitment of metal ions by the enzyme to provide electrophilic catalysis of enolate formation. PMID- 11902888 TI - Two-dimensional (2D) pulsed electron paramagnetic resonance study of VO(2+) triphosphate interactions: evidence for tridentate triphosphate coordination, and relevance to bone uptake and insulin enhancement by vanadium pharmaceuticals. AB - Two- and four-pulse electron spin echo envelope modulation (ESEEM) and four-pulse two-dimensional hyperfine sublevel correlation (HYSCORE) spectroscopies have been used to determine the solution structure of a 3:1 triphosphate:vanadyl solution at pH 5.0. Limited quantitative data were extracted from the two pulse spectra; however, HYSCORE proved to be more useful in the detection and interpretation of the (31)P and (1)H couplings. Three sets of cross-peaks were observed for each nucleus. For the (31)P couplings, three sets of cross-peaks were observed in the HYSCORE spectrum, and contour line shape analysis yielded coupling constants of approximately 15, 9, and 1 MHz. HYSCORE cross-peaks in the proton region were partially overlapping; however, interpretation of the proton coupling was simplified through the use of one-dimensional four-pulse ESEEM and subsequent analysis of the sum combination peaks. Comparison of the derived isotropic and anisotropic coupling constants with results from earlier ESEEM and electron nuclear double resonance (ENDOR) studies was consistent with the presence of at least one, and most likely two, water molecules coordinated in the equatorial plane of the vanadyl cation. The vanadyl-triphosphate system was shown to be an accurate model of the in vivo vanadyl-phosphate coupling constants determined in an earlier study (Dikanov, S. A.; Liboiron, B. D.; Thompson, K. H.; Vera, E.; Yuen, V. G.; McNeill, J. H.; Orvig, C. J. Am. Chem. Soc. 1999, 121, 11004.) Comparison of these values to those found in previous spectroscopic studies of vanadyl-triphosphate interactions, along with a detailed structural interpretation, are presented. This work represents the first detection of tridentate polyphosphate coordination to the vanadyl ion, and the first observation of an axial phosphate interaction not previously reported in earlier ENDOR and pulsed electron paramagnetic resonance studies. PMID- 11902889 TI - Solution synthesis of gadolinium nanoparticles. AB - Gadolinium nanoparticles have been produced at subambient temperature by alkalide reduction. The nanoparticles display maxima in the temperature dependence of their magnetization, cooled in the absence of an applied external field, at T(max) of 5.0 and 17.5 K for unheated samples and samples annealed at 1000 degrees C for 4 h, respectively. Field cooled behavior deviates at temperatures slightly above T(max), increasing at lower temperature. Curie-Weiss law fits of the high-temperature data yield magnetic moments in close agreement with those expected for noninteracting Gd(3+) ions, suggesting that the behavior seen is due to a magnetic transition rather than superparamagnetism. Magnetization is linearly dependent on field at temperatures higher than 7-8 times T(max) and shows remanence-free hysteresis at lower temperature, suggesting metamagnetism. Some annealed samples show evidence of additional ferromagnetic interactions below approximately 170 K. Magnetic entropy curves generated from magnetization data are consistent with that expected for a paramagnet. PMID- 11902890 TI - Comprehensive thermodynamic characterization of the metal-hydrogen bond in a series of cobalt-hydride complexes. AB - A detailed structural and thermodynamic study of a series of cobalt-hydride complexes is reported. This includes structural studies of [H(2)Co(dppe)(2)](+), HCo(dppe)(2), [HCo(dppe)(2)(CH(3)CN)](+), and [Co(dppe)(2)(CH(3)CN)](2+), where dppe = bis(diphenylphosphino)ethane. Equilibrium measurements are reported for one hydride- and two proton-transfer reactions. These measurements and the determinations of various electrochemical potentials were used to determine 11 of 12 possible homolytic and heterolytic solution Co-H bond dissociation free energies of [H(2)Co(dppe)(2)](+) and its monohydride derivatives. These values provide a useful framework for understanding observed and potential reactions of these complexes. These reactions include the disproportionation of [HCo(dppe)(2)](+) to form [Co(dppe)(2)](+) and [H(2)Co(dppe)(2)](+), the reaction of [Co(dppe)(2)](+) with H(2), the protonation and deprotonation reactions of the various hydride species, and the relative ability of the hydride complexes to act as hydride donors. PMID- 11902891 TI - Supramolecular chirogenesis in zinc porphyrins: equilibria, binding properties, and thermodynamics. AB - Complexation mechanism, binding properties and thermodynamic parameters of supramolecular chirality induction in the achiral host molecule, syn (face-to face conformation) ethane-bridged bis(zinc porphyrin), upon interaction with chiral monoamine and monoalcohol guests have been studied by means of the UV-vis, CD, (1)H NMR, and ESI MS techniques. It was found that the chirogenesis process includes three major equilibria steps: the first guest ligation to a zinc porphyrin subunit of the host (K(1)), syn to anti conformational switching (K(S)), and further ligation by a second guest molecule to the remaining ligand free zinc porphyrin subunit (K(2)), thus forming the final bis-ligated species possessing supramolecular chirality. The validity of this equilibria model is confirmed by the excellent match between the calculated and experimentally observed spectral parameters of the bis-ligated species. The second ligation proceeds in a cooperative manner as K(2) > K(1) for all supramolecular systems studied, regardless of the structure of the chiral ligand used. The binding properties are highly dependent on the nature of the functional group (amines are stronger binders than alcohols) and on the structure of the chiral guests (primary and aliphatic amines have overall binding constant values greater than those of secondary and aromatic amines, respectively). PMID- 11902892 TI - Metastable isonitrosyl structure of the nitroprusside anion confirmed by nuclear inelastic scattering. AB - Nuclear inelastic scattering (NIS) measurements were performed on a guanidium nitroprusside ((CN(3)H(6))(2)[Fe(CN)(5)NO], GNP) monocrystal at 77 K after the sample was illuminated with blue light (450 nm) at 50 K to populate the two metastable states, MS(1) and MS(2), of the nitroprusside anion. A second measurement was performed at 77 K after warming up the illuminated crystal to 250 K where the metastable states decay to the groundstate. The measured spectra were compared with simulated NIS spectra that were calculated by using density functional methods. Comparison of measured and simulated spectra provides strong evidence for the isonitrosyl structure of the metastable MS(1) state proposed by Carducci et al. (Carducci, M. D.; Pressprich, M. R.; Coppens, P. J. Am. Chem. Soc. 1997, 119, 2669-2678). PMID- 11902893 TI - Planar three-coordinate high-spin Fe(II) complexes with large orbital angular momentum: Mossbauer, electron paramagnetic resonance, and electronic structure studies. AB - Mossbauer spectra of [LFe(II)X](0) (L = beta-diketiminate; X = Cl(-), CH(3)(-), NHTol(-), NHtBu(-)), 1.X, were recorded between 4.2 and 200 K in applied magnetic fields up to 8.0 T. A spin Hamiltonian analysis of these data revealed a spin S = 2 system with uniaxial magnetization properties, arising from a quasi-degenerate M(S) = +/-2 doublet that is separated from the next magnetic sublevels by very large zero-field splittings (3/D/ > 150 cm(-1)). The ground levels give rise to positive magnetic hyperfine fields of unprecedented magnitudes, B(int) = +82, +78, +72, and +62 T for 1.CH(3), 1.NHTol, 1.NHtBu, and 1.Cl, respectively. Parallel-mode EPR measurements at X-band gave effective g values that are considerably larger than the spin-only value 8, namely g(eff) = 10.9 (1.Cl) and 11.4 (1.CH(3)), suggesting the presence of unquenched orbital angular momenta. A qualitative crystal field analysis of g(eff) shows that these momenta originate from spin-orbit coupling between energetically closely spaced yz and z(2) 3d orbital states at iron and that the spin of the M(S) = +/-2 doublet is quantized along x, where x is along the Fe-X vector and z is normal to the molecular plane. A quantitative analysis of g(eff) provides the magnitude of the crystal field splitting of the lowest two orbitals, /epsilon(yz) - epsilon(2)(z)/ = 452 (1.Cl) and 135 cm(-1) (1.CH(3)). A determination of the sign of the crystal field splitting was attempted by analyzing the electric field gradient (EFG) at the (57)Fe nuclei, taking into account explicitly the influence of spin-orbit coupling on the valence term and ligand contributions. This analysis, however, led to ambiguous results for the sign of epsilon(yz) - epsilon(2)(z). The ambiguity was resolved by analyzing the splitting Delta of the M(S) = +/-2 doublet; Delta = 0.3 cm(-1) for 1.Cl and Delta = 0.03 cm(-)(1) for 1.CH(3). This approach showed that z(2) is the ground state in both complexes and that epsilon(yz) - epsilon(2)(z) approximately 3500 cm(-1) for 1.Cl and 6000 cm(-1) for 1.CH(3). The crystal field states and energies were compared with the results obtained from time-dependent density functional theory (TD-DFT). The isomer shifts and electric field gradients in 1.X exhibit a remarkably strong dependence on ligand X. The ligand contributions to the EFG, denoted W, were expressed by assigning ligand-specific parameters: W(X) to ligands X and W(N) to the diketiminate nitrogens. The additivity and transferability hypotheses underlying this model were confirmed by DFT calculations. The analysis of the EFG data for 1.X yields the ordering W(N(diketiminate)) < W(Cl) < W(N'HR), W(CH(3)) and indicates that the diketiminate nitrogens perturb the iron wave function to a considerably lesser extent than the monodentate nitrogen donors do. Finally, our study of these synthetic model complexes suggests an explanation for the unusual values for the electric hyperfine parameters of the iron sites in the Fe-Mo cofactor of nitrogenase in the M(N) state. PMID- 11902895 TI - Structural analysis of the solid amorphous binuclear complexes of iron(III) and aluminum(III) with chromium(III)-DTPA chelator using energy dispersive X-ray diffraction. AB - This paper is concerned with the structural data obtained for two amorphous binuclear complexes of iron(III) and aluminum(III) with chromium(III) diethylentriaminepentaacetic acid (chromium(III)-DTPA, CrL(2)(-)) using the energy-dispersive X-ray diffraction technique. Fe(OH)CrL(H(2)O)(6) and Al(OH)CrL(H(2)O)(6) are binuclear complexes, the metals ions being bridged via oxygen atoms. The metal ions are all octahedrally coordinated. PMID- 11902894 TI - Olefin cis-dihydroxylation versus epoxidation by non-heme iron catalysts: two faces of an Fe(III)-OOH coin. AB - The oxygenation of carbon-carbon double bonds by iron enzymes generally results in the formation of epoxides, except in the case of the Rieske dioxygenases, where cis-diols are produced. Herein we report a systematic study of olefin oxidations with H(2)O(2) catalyzed by a group of non-heme iron complexes, i.e., [Fe(II)(BPMEN)(CH(3)CN)(2)](2+) (1, BPMEN = N,N'-dimethyl-N,N'-bis(2 pyridylmethyl)-1,2-diaminoethane) and [Fe(II)(TPA)(CH(3)CN)(2)](2+) (4, TPA = tris(2-pyridylmethyl)amine) and their 6- and 5-methyl-substituted derivatives. We demonstrate that olefin epoxidation and cis-dihydroxylation are different facets of the reactivity of a common Fe(III)-OOH intermediate, whose spin state can be modulated by the electronic and steric properties of the ligand environment. Highly stereoselective epoxidation is favored by catalysts with no more than one 6-methyl substituent, which give rise to low-spin Fe(III)-OOH species (category A). On the other hand, cis-dihydroxylation is favored by catalysts with more than one 6-methyl substituent, which afford high-spin Fe(III)-OOH species (category B). For catalysts in category A, both the epoxide and the cis-diol product incorporate (18)O from H(2)(18)O, results that implicate a cis-H(18)O-Fe(V)=O species derived from O-O bond heterolysis of a cis-H(2)(18)O-Fe(III)-OOH intermediate. In contrast, catalysts in category B incorporate both oxygen atoms from H(2)(18)O(2) into the dominant cis-diol product, via a putative Fe(III) eta(2)-OOH species. Thus, a key feature of the catalysts in this family is the availability of two cis labile sites, required for peroxide activation. The olefin epoxidation and cis-dihydroxylation studies described here not only corroborate the mechanistic scheme derived from our earlier studies on alkane hydroxylation by this same family of catalysts (Chen, K.; Que, L, Jr. J. Am. Chem. Soc. 2001, 123, 6327) but also further enhance its credibility. Taken together, these reactions demonstrate the catalytic versatility of these complexes and provide a rationale for Nature's choice of ligand environments in biocatalysts that carry out olefin oxidations. PMID- 11902896 TI - Deuterium structural effects in inorganic and bioinorganic aggregates. AB - Deuterium kinetic isotope effects are widely used in chemical and biological research. Deuterium thermodynamic effects on the aqueous synthesis of inorganic materials, however, seem not to have been recognized. Here we report that the simple replacement of H(2)O with D(2)O in the synthesis of a solid-state manganese complex results in a new structurally and magnetically distinct phase. When iron oxides are synthesized, the relative amount of the mineral phases obtained in H(2)O vs D(2)O is different. The morphology and magnetic properties of the iron core of the iron storage protein ferritin are likewise different when mineralization is carried out in heavy water. The formation of extra inorganic solids, change in the ratio of two phases or alteration of a single phase morphology in D(2)O suggest that new inorganic and bioinorganic metal complexes might be obtained by using the thermodynamic isotope effect. PMID- 11902897 TI - Oxalate-bridged complexes of dimolybdenum and ditungsten supported by pivalate ligands: ((t)BuCO(2))(3)M(2)(mu-O(2)CCO(2))M(2)(O(2)C(t)Bu)(3). Correlation of the solid-state, molecular, and electronic structures with Raman, resonance Raman, and electronic spectral data. AB - The compounds ((t)BuCO(2))(3)M(2)(mu-O(2)CCO(2))M(2)(O(2)C(t)Bu)(3) (M(4)OXA), where M = Mo or W, are shown by analysis of powder X-ray diffraction data to have extended lattice structures wherein oxygen atoms from the oxalate and pivalate ligands of one M(4)OXA molecule are linked to metal atoms of neighboring molecules. Raman, resonance Raman, electronic absorption (2-325 K in 2-MeTHF), and emission spectra are reported, together with corresponding spectra of the mu O(2)(13)C(13)CO(2) isotopomers. To aid in the assignment, the Raman spectra of K(2)C(2)O(4).H(2)O and K(2)(13)C(2)O(4).H(2)O have also been recorded. The visible region of the electronic spectra is dominated by intense, fully allowed MLCT transitions, M(2) delta to oxalate pi*, which show pronounced thermochromism and extensive vibronic progressions associated with the oxalate ligand at low temperatures. With excitation into these charge-transfer bands, strong resonance enhancement is seen for Raman bands assigned to the oxalate nu(1)(a(g)) and, to a lesser extent, nu(2)(a(g)) modes. Electronic structure calculations for the model compounds (HCO(2))(3)M(2)(mu-O(2)CCO(2))M(2)(O(2)CH)(3), employing density functional theory (gradient corrected and time-dependent) with the Gaussian 98 and ADF 2000 packages, predict the planar oxalate D(2h) configuration to be favored, which maximizes M(2) delta to oxalate pi* back-bonding, and indicate low barriers (<8 kcal mol(-1)) to rotation about the oxalate C-C bonds. PMID- 11902898 TI - Organometallic ruthenium(II) diamine anticancer complexes: arene-nucleobase stacking and stereospecific hydrogen-bonding in guanine adducts. AB - Organometallic ruthenium(II) arene anticancer complexes of the type [(eta(6) arene)Ru(II)(en)Cl][PF(6)] (en = ethylenediamine) specifically target guanine bases of DNA oligomers and form monofunctional adducts (Morris, R., et al. J. Med. Chem. 2001). We have determined the structures of monofunctional adducts of the "piano-stool" complexes [(eta(6)-Bip)Ru(II)(en)Cl][PF(6)] (1, Bip = biphenyl), [(eta(6)-THA)Ru(II)(en)Cl][PF(6)] (2, THA = 5,8,9,10 tetrahydroanthracene), and [(eta(6)-DHA)Ru(II)(en)Cl][PF(6)] (3, DHA = 9,10 dihydroanthracene) with guanine derivatives, in the solid state by X-ray crystallography, and in solution using 2D [(1)H,(1)H] NOESY and [(1)H,(15)N] HSQC NMR methods. Strong pi-pi arene-nucleobase stacking is present in the crystal structures of [(eta(6)-C(14)H(14))Ru(en)(9EtG-N7)][PF(6)](2).(MeOH) (6) and [(eta(6)-C(14)H(12))Ru(en)(9EtG-N7)][PF(6)](2).2(MeOH) (7) (9EtG = 9 ethylguanine). The anthracene outer ring (C) stacks over the purine base at distances of 3.45 A for 6 and 3.31 A for 7, with dihedral angles of 3.3 degrees and 3.1 degrees, respectively. In the crystal structure of [(eta(6) biphenyl)Ru(en)(9EtG-N7)][PF(6)](2).(MeOH) (4), there is intermolecular stacking between the pendant phenyl ring and the purine six-membered ring at a distance of 4.0 A (dihedral angle 4.5 degrees). This stacking stabilizes a cyclic tetramer structure in the unit cell. The guanosine (Guo) adduct [(eta(6) biphenyl)Ru(en)(Guo-N7)][PF(6)](2).3.75(H(2)O) (5) exhibits intramolecular stacking of the pendant phenyl ring with the purine five-membered ring (3.8 A, 23.8 degrees) and intermolecular stacking of the purine six-membered ring with an adjacent pendant phenyl ring (4.2 A, 23.0 degrees). These occur alternately giving a columnar-type structure. A syn orientation of arene and purine is present in the crystal structures 5, 6, and 7, while the orientation is anti for 4. However, in solution, a syn orientation predominates for all the biphenyl adducts 4, 5, and the guanosine 5'-monophosphate (5'-GMP) adduct 8 [(eta(6) biphenyl)Ru(II)(en)(5'-GMP-N7)], as revealed by NMR NOE studies. The predominance of the syn orientation both in the solid state and in solution can be attributed to hydrophobic interactions between the arene and purine rings. There are significant reorientations and conformational changes of the arene ligands in [(eta(6)-arene)Ru(II)(en)(G-N7)] complexes in the solid state, with respect to those of the parent chloro-complexes [(eta(6)-arene)Ru(II)(en)Cl](+). The arene ligands have flexibility through rotation around the arene-Ru pi-bonds, propeller twisting for Bip, and hinge-bending for THA and DHA. Thus propeller twisting of Bip decreases by ca. 10 degrees so as to maximize intra- or intermolecular stacking with the purine ring, and stacking of THA and DHA with the purine is optimized when their tricyclic ring systems are bent by ca. 30 degrees, which involves increased bending of THA and a flattening of DHA. This flexibility makes simultaneous arene-base stacking and N7-covalent binding compatible. Strong stereospecific intramolecular H-bonding between an en NH proton oriented away from the arene (en NH(d)) and the C6 carbonyl of G (G O6) is present in the crystal structures of 4, 5, 6, and 7 (average N...O distance 2.8 A, N-H...O angle 163 degrees ). NMR studies of the 5'-GMP adduct 8 provided evidence that en NH(d) protons are involved in strong H-bonding with the 5'-phosphate and O6 of 5'-GMP. The strong H-bonding from G O6 to en NH(d) protons partly accounts for the high preference for binding of [(eta(6)-arene)Ru(II)en](2+) to G versus A (adenine). These studies suggest that simultaneous covalent coordination, intercalation, and stereospecific H-bonding can be incorporated into Ru(II) arene complexes to optimize their DNA recognition behavior, and as potential drug design features. PMID- 11902900 TI - Quantum mechanics/molecular mechanics studies of triosephosphate isomerase catalyzed reactions: effect of geometry and tunneling on proton-transfer rate constants. AB - The role of tunneling for two proton-transfer steps in the reactions catalyzed by triosephosphate isomerase (TIM) has been studied. One step is the rate-limiting proton transfer from Calpha in the substrate to Glu 165, and the other is an intrasubstrate proton transfer proposed for the isomerization of the enediolate intermediate. The latter, which is not important in the wild-type enzyme but is a useful model system because of its simplicity, has also been examined in the gas phase and in solution. Variational transition-state theory with semiclassical ground-state tunneling was used for the calculation with potential energy surface determined by an AM1 method specifically parametrized for the TIM system. The effect of tunneling on the reaction rate was found to be less than a factor of 10 at room temperature; the tunneling becomes more important at lower temperature, as expected. The imaginary frequency (barrier) mode and modes that have large contributions to the reaction path curvature are localized on the atoms in the active site, within 4 A of the substrate. This suggests that only a small number of atoms that are close to the substrate and their motions (e.g., donor-acceptor vibration) directly determine the magnitude of tunneling. Atoms that are farther away influence the effect of tunneling indirectly by modulating the energetics of the proton transfer. For the intramolecular proton transfer, tunneling was found to be most important in the gas phase, to be similar in the enzyme, and to be the smallest in water. The major reason for this trend is that the barrier frequency is substantially lower in solution than in the gas phase and enzyme; the broader solution barrier is caused by the strong electrostatic interaction between the highly charged solute and the polar solvent molecules. Analysis of isotope effects showed that the conventional Arrenhius parameters are more useful as experimental criteria for determining the magnitude of tunneling than the widely used Swain-Schaad exponent (SSE). For the primary SSE, although values larger than the transition-state theory limit (3.3) occur when tunneling is included, there is no clear relationship between the calculated magnitudes of tunneling and the SSE. Also, the temperature dependence of the primary SSE is rather complex; the value of SSE tends to decrease as the temperature is lowered (i.e., when tunneling becomes more significant). For the secondary SSE, the results suggest that it is more relevant for evaluating the "coupled motion" between the secondary hydrogen and the reaction coordinate than the magnitude of tunneling. Although tunneling makes a significant contribution to the rate of proton transfer, it appears not to be a major aspect of the catalysis by TIM at room temperature; i.e., the tunneling factor of 10 is "small" relative to the overall rate acceleration by 10(9). For the intramolecular proton transfer, the tunneling in the enzyme is larger by a factor of 5 than in solution. PMID- 11902899 TI - Structural characterization of metallopeptides designed as scaffolds for the stabilization of nickel(II)-Fe(4)S(4) bridged assemblies by X-ray absorption spectroscopy. AB - In earlier work, de novo designed peptides with a helix-loop-helix motif and 63 residues have been synthesized as potential scaffolds for stabilization of the [Ni(II)-X-Fe(4)S(4)] bridged assembly that is the spectroscopically deduced structure of the A-Cluster in clostridial carbon monoxide dehydrogenase. The 63mers contain a consensus tricysteinyl ferredoxin domain in the loop for binding an Fe(4)S(4) cluster and Cys and His residues proximate to the loop for binding Ni(II), with one Cys residue designed as the bridge X. The metallopeptides HC(4)H(2)-[Fe(4)S(4)]-Ni and HC(5)H-[Fe(4)S(4)]-M, containing three His and one Cys residue for Ni(II) coordination and two His and two Cys residues for binding M = Ni(II) and Co(II), have been examined by Fe-, Ni-, and Co-K edge spectroscopy and EXAFS. All peptides bind an [Fe(4)S(4)](2+) cubane-type cluster. Interpretation of the Ni and Co data is complicated by the presence of a minority population of six-coordinate species with low Z ligands, designated for simplicity as [M(OH(2))(6)](2+). Best fits of the data were obtained with ca. 20% [M(OH(2))(6)](2+) and ca. 80% M(II) with mixed N/S coordination. The collective XAS results for HC(4)H(2)-[Fe(4)S(4)]-Ni and HC(5)H-[Fe(4)S(4)]-M demonstrate the presence of an Fe(4)S(4) cluster and support the existence of the distorted square-planar coordination units [Ni(II)(S.Cys)(N.His)(3)] and [Ni(II)(S.Cys)(2)(N.His)(2)] in the HC(4)H(2) and HC(5)H metallopeptides, respectively. In the HC(5)H metallopeptide, tetrahedral [Co(II)(S.Cys)(2)(N.His)(2)] is present. We conclude that the designed scaffolded binding sites, including Ni-(mu(2)-S.Cys)-Fe bridges, have been achieved. This is the first XAS study of a de novo designed metallopeptide intended to stabilize a bridged biological assembly, and one of a few XAS analyses of metal derivatives of designed peptides. The scaffolding concept should be extendable to other bridged metal assemblies. PMID- 11902901 TI - Multiple quantum filtered NMR studies of the interaction between collagen and water in the tendon. AB - We studied the physical processes and the chemical reactions involved in magnetization transfer between water and large proteins, such as collagen, in bovine Achilles tendon. Since the NMR spectrum for such proteins is broadened by very large dipolar interactions, the NMR peaks of the various functional groups on the protein cannot be separated from one another on the basis of their different chemical shifts. A further complication in observing the protein spectrum is the intense narrow peak of the abundant water. Thus, magnetization transfer (MT) within the protein or between water and the protein cannot rely on differences in the chemical shifts, as is commonly possible in liquids. We present a method that separates the protein spectrum from that of the water spectrum on the basis of their different intramolecular dipolar interactions, enabling exclusive excitation of either the protein or water. As a result, the protein spectrum as well as the effect of spin diffusion within the protein can be measured. In addition, the MT rates from the protein to water and vice versa can be measured. Two types of mechanisms were considered for the MT: chemical exchange- and dipolar interaction-related processes (such as NOE). They were distinguished by examining the effects of the following experimental conditions: (a) temperature; (b) pH; (c) ratio of D(2)O to H(2)O in the bathing liquid; (d) interaction of the protein with small molecules other than water, such as DMSO and methanol. Our results lead us to the conclusion that the MT is dominated below the freezing point by the dipolar interaction between the protein and water, while an exchange of protons between the protein and the water molecules is the most significant process above the freezing point. On the basis of the fact that the spin temperature is established for the protein on a time scale much shorter than that of the MT, we could measure protein spectra that are distinguished by the contributions made to them by the various functional groups; i.e., contributions of methylenes were distinguished from those of methyls. PMID- 11902902 TI - Hoogsteen-based parallel-stranded duplexes of DNA. Effect of 8-amino-purine derivatives. AB - The structure of parallel-stranded duplexes of DNA-containing a mixture of guanines (G) and adenines (A) is studied by means of molecular dynamics (MD) simulation, as well as NMR and circular dichroism (CD) spectroscopy. Results demonstrate that the structure is based on the Hoogsteen motif rather than on the reverse Watson-Crick one. Molecular dynamics coupled to thermodynamic integration (MD/TI) calculations and melting experiments allowed us to determine the effect of 8-amino derivatives of A and G and of 8-amino-2'-deoxyinosine on the stability of parallel-stranded duplexes. The large stabilization of the parallel-stranded helix upon 8-amino substitution agrees with a Hoogsteen pairing, confirming MD, NMR, and CD data, and suggests new methods to obtain DNA triplexes for antigene and antisense purposes. PMID- 11902903 TI - Electron-nuclear double resonance spectroscopic evidence that S adenosylmethionine binds in contact with the catalytically active [4Fe-4S](+) cluster of pyruvate formate-lyase activating enzyme. AB - Pyruvate formate-lyase activating enzyme (PFL-AE) is a representative member of an emerging family of enzymes that utilize iron-sulfur clusters and S adenosylmethionine (AdoMet) to initiate radical catalysis. Although these enzymes have diverse functions, evidence is emerging that they operate by a common mechanism in which a [4Fe-4S](+) interacts with AdoMet to generate a 5' deoxyadenosyl radical intermediate. To date, however, it has been unclear whether the iron-sulfur cluster is a simple electron-transfer center or whether it participates directly in the radical generation chemistry. Here we utilize electron paramagnetic resonance (EPR) and pulsed 35 GHz electron-nuclear double resonance (ENDOR) spectroscopy to address this question. EPR spectroscopy reveals a dramatic effect of AdoMet on the EPR spectrum of the [4Fe-4S](+) of PFL-AE, changing it from rhombic (g = 2.02, 1.94, 1.88) to nearly axial (g = 2.01, 1.88, 1.87). (2)H and (13)C ENDOR spectroscopy was performed on [4Fe-4S](+)-PFL-AE (S = (1)/(2)) in the presence of AdoMet labeled at the methyl position with either (2)H or (13)C (denoted [1+/AdoMet]). The observation of a substantial (2)H coupling of approximately 1 MHz ( approximately 6-7 MHz for (1)H), as well as hyperfine-split signals from the (13)C, manifestly require that AdoMet lie close to the cluster. (2)H and (13)C ENDOR data were also obtained for the interaction of AdoMet with the diamagnetic [4Fe-4S](2+) state of PFL-AE, which is visualized through cryoreduction of the frozen [4Fe-4S](2+)/AdoMet complex to form the reduced state (denoted [2+/AdoMet](red)) trapped in the structure of the oxidized state. (2)H and (13)C ENDOR spectra for [2+/AdoMet](red) are essentially identical to those obtained for the [1+/AdoMet] samples, showing that the cofactor binds in the same geometry to both the 1+ and 2+ states of PFL-AE. Analysis of 2D field-frequency (13)C ENDOR data reveals an isotropic hyperfine contribution, which requires that AdoMet lie in contact with the cluster, weakly interacting with it through an incipient bond/antibond. From the anisotropic hyperfine contributions for the (2)H and (13)C ENDOR, we have estimated the distance from the closest methyl proton of AdoMet to the closest iron of the cluster to be approximately 3.0-3.8 A, while the distance from the methyl carbon to the nearest iron is approximately 4-5 A. We have used this information to construct a model for the interaction of AdoMet with the [4Fe-4S](2+/+) cluster of PFL-AE and have proposed a mechanism for radical generation that is consistent with these results. PMID- 11902905 TI - Cyclic carbon cluster dianions and their aromaticity. AB - Cyclic carbon cluster dianions (CC(2))(2-)(n)(n = 3-6) are investigated by ab initio methods with regard to their geometric properties, electronic stability, and aromaticity. The unique wheel-like structures of these dianions consist of a n-membered carbon ring, where a C(2) unit is attached to each carbon atom. All investigated dianions represent stable gas-phase dianions. While the smallest member of this family (CC(2))(2-)(3) is clearly aromatic, the aromatic character decreases rapidly with increasing ring size. The geometries and the aromaticity of the cyclic clusters (CC(2))(2-)(n)(n = 3-6) can be nicely explained using resonance structure arguments. PMID- 11902904 TI - The interaction of MS-325 with human serum albumin and its effect on proton relaxation rates. AB - MS-325 is a novel blood pool contrast agent for magnetic resonance imaging currently undergoing clinical trials to assess blockage in arteries. MS-325 functions by binding to human serum albumin (HSA) in plasma. Binding to HSA serves to prolong plasma half-life, retain the agent in the blood pool, and increase the relaxation rate of water protons in plasma. Ultrafiltration studies with a 5 kDa molecular weight cutoff filter show that MS-325 binds to HSA with stepwise stoichiometric affinity constants (mM(-1)) of K(a1) = 11.0 +/- 2.7, K(a2) = 0.84 +/- 0.16, K(a3) = 0.26 +/- 0.14, and K(a4) = 0.43 +/- 0.24. Under the conditions 0.1 mM MS-325, 4.5% HSA, pH 7.4 (phosphate-buffered saline), and 37 degrees C, 88 +/- 2% of MS-325 is bound to albumin. Fluorescent probe displacement studies show that MS-325 can displace dansyl sarcosine and dansyl-L asparagine from HSA with inhibition constants (K(i)) of 85 +/- 3 microM and 1500 +/- 850 microM, respectively; however, MS-325 is unable to displace warfarin. These results suggest that MS-325 binds primarily to site II on HSA. The relaxivity of MS-325 when bound to HSA is shown to be site dependent. The Eu(III) analogue of MS-325 is shown to contain one inner-sphere water molecule in the presence and in the absence of HSA. The synthesis of an MS-325 analogue, 5, containing no inner-sphere water molecules is described. Compound 5 is used to estimate the contribution to relaxivity from the outer-sphere water molecules surrounding MS-325. The high relaxivity of MS-325 bound to HSA is primarily because of a 60-100-fold increase in the rotational correlation time of the molecule upon binding (tau(R) = 10.1 +/- 2.6 ns bound vs 115 ps free). Analysis of the nuclear magnetic relaxation dispersion (T(1) and T(2)) profiles also suggests a decrease in the electronic relaxation rate (1/T(1e) at 20 MHz = 2.0 x 10(8) s(-1) bound vs 1.1 x 10(9) s(-1) free) and an increase in the inner-sphere water residency time (tau(m) = 170 +/- 40 ns bound vs 69 +/- 20 ns free). PMID- 11902906 TI - High resolution capillary electrophoresis of carbon nanotubes. AB - Purification of single-walled carbon nanotubes by capillary electrophoresis (CE) is demonstrated. Real-time Raman spectroscopy of the separation process and single-wavelength UV/vis detection show the ability of CE to provide high resolution separations of nanotube fractions with baseline separation. AFM images of collected fractions demonstrate that separations are based on tube length. The separation method is suggested to be based on alignment of the nanotubes along the separation field. PMID- 11902907 TI - Stereoelectronic effects and general trends in hyperconjugative acceptor ability of sigma bonds. AB - A systematic study of general trends in sigma acceptor properties of C-X bonds where X is a main group element from groups IVa-IIa is presented. The acceptor ability of the C-X sigma bonds in monosubstituted ethanes increases when going to the end of a period and down a group. Enhancement of acceptor ability of C-X sigma bonds as one moves from left to right in periods parallels the increase in electronegativity of X, whereas augmentation of acceptor ability in groups is opposite to the changes in electronegativity of X and in the C-X bond polarization, following instead the decrease in the energy of sigma(C)(-)(X) orbitals when one moves from the top to the bottom within a group. This simple picture of acceptor ability of sigma bonds being controlled by electronegativity in periods and by sigma orbital energy in groups is changed in monosubstituted ethenes where the role of electronegativity of the substituent X becomes more important due to increased overlap between sigma orbitals. The combination of several effects of similar magnitude influences acceptor ability of sigma bonds in monosubstituted ethenes in a complex way. As a result, the acceptor ability of sigma bonds can be significantly modified by substitution and is conformer dependent. Stereoelectronic effects displayed by C-X bonds with X from second and third periods are highly anisotropic. For example, C-chalcogen bonds are excellent sigma acceptors at the carbon end but poor sigma acceptors at the chalcogen end. This effect can be relied upon in the design of molecular diodes with sigma bridges with unidirectional electron conductivity. While the general trends revealed in this work should be useful for the qualitative understanding of stereoelectronic effects, one should bear in mind that the magnitude of hyperconjugative effects is extremely sensitive to small variations in structure and in substitution. This advocates for the increased role of theoretical methods in analysis of stereoelectronic effects. PMID- 11902908 TI - Dioxins in food: a modern agricultural perspective. AB - This review attempts to cover and summarize the literature available on polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in the environment with regard to problems of interest to agriculture. The coverage of the literature is extensive (120 references) but, by all means, not complete. Issues that are addressed in this review include a background summary of dioxins in the environment and their potential human health risks; current knowledge on the levels of dioxins in the U.S. food supply and comparisons to European data; descriptions of recent food contamination episodes; an evaluation of methods that may reduce incurred levels of dioxins in livestock and meats; and the status and limitations of dioxin analysis and rapid screening methods with regard to widespread monitoring programs. Research areas in agriculture where data and experimental results are scarce or nonexistent are also pointed out. PMID- 11902909 TI - Recent developments in food characterization and adulteration detection: technique-oriented perspectives. AB - This review covers mainly publications that appeared in Analytical Abstracts (Royal Society of Chemistry) from January 1990 to February 2001. The number of publications on this topic continues to grow, and during the past three years (1998-2000) about 150 reviews and/or overviews have been published in the area of food. Numerous techniques and food matrices or chemical components are presented and discussed in these reviews. The present review is intentionally limited to eight techniques or classes of techniques and intends to be a "technique by technique" presentation of "what was used" or "what is used" to characterize food products and to detect their possible adulteration. The present review focuses on the following techniques: microscopic analysis; HPLC; GC, GC-(MS, FTIR); UV visible spectrophotometry; AAS/AES, ICP-(AES, MS); IRMS, GC-IRMS, GC-C-IRMS; DSC; IR, mid-IR, and NMR (202 references). Emphasis is placed as much as possible on chemometrical treatment of analytical data, which are commonly used to achieve the final objective, either food characterization or adulteration detection. Finally, a brief description is given of the new generation of analytical systems that combine powerful analytical techniques and powerful computer software for a best extraction of the information from analytical data. PMID- 11902910 TI - Development of a CE method to analyze organic acids in dairy products: application to study the metabolism of heat-shocked spores. AB - Organic acids are relevant in dairy products for nutritional reasons and because they contribute to the flavor and aroma. They are the major products of carbohydrate catabolism of lactic acid bacteria and nonstarter bacteria associated with milk. In several research and quality programs, it is very important to develop a rapid and sensitive method for their quantitative determination in dairy products to monitor bacterial growth and activity. A capillary electrophoresis method for the simultaneous determination of oxalic, citric, formic, succinic, orotic, uric, pyruvic, acetic, propionic, lactic, and butyric acid in less than 18 min has been developed. Various parameters affecting analysis, including capillary length, type, composition, and pH of the electrolyte have been optimized. Some alternatives are given to improve the separation of particular organic acids of special interest. Its application to analyze the quality of some dairy products has been investigated. In addition, the suitability of the technique to determine profiles of organic acids generated during the metabolism of heat-shocked spores has been demonstrated. PMID- 11902911 TI - Quantification of exopolysaccharide, lactic acid, and lactose concentrations in culture broth by near-infrared spectroscopy. AB - Near-infrared spectroscopy (NIRS) was used for the simultaneous prediction of exopolysaccharide (EPS; 0-3 g/L) and lactic acid (0-59 g/L) productions as well as lactose (0-68 g/L) concentration in supernatant samples from pH-controlled batch cultures of Lactobacillus rhamnosus RW-9595M in supplemented whey permeate medium. To develop calibration equations, the correlation between the second derivative of 164 NIRS transmittance spectra and concentration data obtained with reference methods was calculated at the wavelength between 1653-1770 and 2041 2353 nm, using a partial least-squares method (PLS). The lactic acid and lactose concentrations were measured by HPLC, and the EPS concentration was estimated by a new ultrafiltration method. The PLS correlation coefficient (R(2)) and the standard error of cross-validation for the calibrations were 91% and 0.26 g/L for EPS, 99% and 2.54 g/L for lactic acid, and 98% and 3.32 g/L for lactose, respectively. The calibration equations were validated with 45 randomly selected culture samples from 6 cultures that were not used for calibration. A high agreement between data of the reference methods and those of NIRS was observed, with correlation coefficients and standard errors of prediction of 99% and 1.64 g/L for lactic acid, 99% and 4.5 g/L for lactose, and 91% and 0.32 g/L for EPS. The results suggest that NIRS could be a useful method for rapid monitoring and control of EPS lactic fermentations. PMID- 11902912 TI - PCR technique for identification of mussel species. AB - Random amplified polymorphic DNA (RAPD) analysis has been applied to the identification of four mussels species: Mytilus edulis, Mytilus chilensis, Mytilus galloprovincialis, and Perna canaliculus. Amplifications of DNA from mussel were carried out using random primers. The most distinctive bands were then isolated, cloned, and sequenced to design specific primers. Finally, DNA from different mussels was amplified with these specific primers, and results allow genetic identification of M. galloprovincialis from the rest of the mussel species. PMID- 11902913 TI - Determination of hydrolyzable tannins (gallotannins and ellagitannins) after reaction with potassium iodate. AB - A widely used method for analyzing hydrolyzable tannins afer reaction with KIO(3) has been modified to include a methanolysis step followed by oxidation with KIO(3). In the new method, hydrolyzable tannins (gallotannins and ellagitannins) are reacted at 85 degrees C for 20 h in methanol/sulfuric acid to quantitatively release methyl gallate. Dried plant samples can be methanolyzed under the same conditions to convert hydrolyzable tannins to methyl gallate. Oxidation of the methyl gallate by KIO(3) at pH 5.5, 30 degrees C, forms a chromophore with lambda(max) 525 nm, which is determined spectrophotometrically. The detection limit of the method is 1.5 microg of methyl gallate, and with plant samples, relative standard deviations of less than 3% were obtained. PMID- 11902915 TI - Advanced solid phase extraction using molecularly imprinted polymers for the determination of quercetin in red wine. AB - Solid phase extraction (SPE) based on molecularly imprinted polymers (MIPs) is a novel approach for sample preparation and preconcentration, gaining increased interest in the fields of environmental, clinical, and food analysis. The first application combining MIPs with SPE for advanced beverage analysis is reported. MIPs for the flavonoid quercetin have been generated, using quercetin as a template molecule in a self-assembly approach and yielding imprinting of 1% of the used template. The MIP achieved a capacity of 0.4 g quercetin per gram polymer and a recovery rate of 98.2%. The application of these synthetic receptors as SPE material for the selective extraction and preconcentration of quercetin from synthetic and red wine samples was investigated. Red wine samples from a French Merlot were directly applied onto the SPE cartridge. The collected fractions were analyzed by high-pressure liquid chromatography. For verification of the obtained results, a similarly prepared nonimprinted polymer and a classical octadecyl silane reversed-phase cartridge were applied as the SPE matrix during control experiments. The MIP enabled the selective extraction of quercetin from a complex matrix, such as red wine, spiked with 8.8 mg per liter quercetin, demonstrating the potential of molecularly imprinted solid phase extraction for rapid, selective, and cost-effective sample pretreatment. PMID- 11902914 TI - Development of an immunoassay for the residues of the herbicide bensulfuron methyl. AB - To develop a competitive indirect enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of the sulfonylurea herbicide bensulfuron methyl, seven structurally related haptens were synthesized. Four of them mimicking the target analyte were conjugated to keyhole limpet hemocyanin by the N-hydroxysuccinimide activated ester method to use as immunogens, and all of them were conjugated to bovine serum albumin to use as plate-coating antigens. Polyclonal antibodies raised in rabbits and the coating antigens were screened and selected for the assay in simple homologous and heterologous ELISA formats. Three sensitive heterologous ELISAs were selected and optimized, showing the average IC(50) values of bensulfuron-methyl as low as 0.17, 0.09, and 0.09 ng/mL, the detection ranges of 0.04-0.60, 0.01-0.60, and 0.04-0.25 ng/mL, and the lowest detection limits of 0.03, 0.002, and 0.03 ng/mL, respectively. The cross reactivities of other sulfonylurea herbicides and metabolites of bensulfuron methyl to the antibodies were less than 15% in the two assays. Recoveries from the analyte-fortified water samples in assay I were in the range of 81-125% by simple dilution. The correlation between the ELISA and HPLC was 0.999 (n = 15) with a slope of 1.37 in the analysis of groundwater samples fortified with bensulfuron-methyl. The results obtained strongly indicate that the ELISA can be a highly sensitive and convenient tool for detecting bensulfuron-methyl residues in agricultural and environmental samples. PMID- 11902917 TI - Development and validation of oxygen radical absorbance capacity assay for lipophilic antioxidants using randomly methylated beta-cyclodextrin as the solubility enhancer. AB - We recently reported the improved oxygen radical absorbance capacity (ORAC) assay using fluorescein (FL) as the fluorescent probe. The current ORAC(FL) assay is limited in hydrophilic antioxidant due to the aqueous environment of the assay. Lipophilic antioxidants mainly include the vitamin E family and carotenoids, which play a critical role in biological defense systems. In this paper, we expanded the current ORAC(FL) assay to lipophilic antioxidants. Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water solubility enhancer for lipophilic antioxidants. Seven percent RMCD (w/v) in a 50% acetone H(2)O mixture was found to sufficiently solubilize vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). This newly developed ORAC assay (abbbreviated ORAC(FL-LIPO)) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrate that the ORAC(FL-LIPO) assay is reliable and robust. For the first time, by using 6-hydroxy-2,5,7,8-tetramethyl-2-carboxylic acid as a standard (1.0), the ORAC values of alpha-tocopherol, (+)-gamma-tocopherol, (+)-delta tocopherol, alpha-tocopherol acetate, tocotrienols, 2,6-di-tert-butyl-4 methylphenol, and gamma-oryzanol were determined to be 0.5 +/- 0.02, 0.74 +/- 0.03, 1.36 +/- 0.14, 0.00, 0.91 +/- 0.04, 0.16 +/- 0.01, and 3.00 +/- 0.26, respectively. The structural information of oxidized alpha-tocopherol obtained by liquid chromatography/mass spectrometry reveals that the mechanism for the reaction between the vitamin E and the peroxyl radical follows the hydrogen atom transfer mechanism, which is in agreement with the notion that vitamin E is the chain-breaking antioxidant. PMID- 11902918 TI - Comparison of gravimetry and hydrolysis/derivatization/gas chromatography-mass spectrometry for quantitative analysis of fat from standard reference infant formula powder using supercritical fluid extraction. AB - This paper describes a comparative study of the gravimetric versus hydrolysis/derivatization/gas chromatography-mass spectrometry determination of fat in infant formula. Fat was extracted using supercritical carbon dioxide modified with a small amount of ethanol, the extract was weighed, and the total fat was determined gravimetrically. Subsequently, another sample of the supercritical fluid fat extract was hydrolyzed to yield free fatty acids, which were converted to their methyl ester derivatives (FAMEs). Quantification was performed by GC-MS. NIST Standard Reference Material (SRM-1846) was used to validate both fat determination methods. Results showed that the gravimetric average percent fat was 26.86%, whereas the GC-MS method yielded 24.64%. Some peaks were detected in the ion chromatogram from the GC-MS that were identified as nonfatty acids such as aldehydes, which may account for the higher percentage fat measured as weight of extract rather than measured as FAMEs expressed as triglycerides. PMID- 11902916 TI - Detection of vinegary defect in virgin olive oils by metal oxide sensors. AB - An array of metal oxide sensors has been set up for detecting the vinegary defect in virgin olive oil. The optimization process was carried out evaluating the variables affecting the process by three desirability functions. Repeatability studies for 6 months and within day were done to evaluate the sensor responses and remove those with high relative standard deviation. The sensor responses were preprocessed applying five weight functions previously to build a regression model. Samples of Spanish Arbequina and Picual virgin olive oil varieties spiked with different amounts of acetic acid (15-200 mg/L) were used as a training set for the regression model. The test set was composed of samples of Italian Coratina virgin olive oil spiked with the vinegary standard at five percentages (10, 25, 40, 50, and 75). A fine-adjusted regression coefficient (R(adj)(2) = 0.98) was computed with the test set. PMID- 11902920 TI - Study of catechin and xanthine tea profiles as geographical tracers. AB - The contents of gallic acid, epigallocatechin gallate, epigallocatechin, epicatechin, epicatechin gallate, catechin, caffeine, theophylline, and theobromine were determined in a set of 45 tea samples, including fermented (black and red) and nonfermented (green) teas of different geographical origins (i.e., China, Japan, Kenya, Sri Lanka, and India). A reversed-phase high performance liquid chromatographic method with gradient elution and photometric detection at 275 nm was used to carry out the analysis. Before the HPLC determination, an extraction step was developed using a mixture of acetonitrile and water (60:40, v/v). Pattern recognition techniques involving principal component analysis (PCA) and linear discriminant analysis (LDA) were applied to differentiate the tea samples according to their geographical origins. Catechins, gallic acid, and tea alkaloids are adequate chemical descriptors to distinguish between fermented and nonfermented tea samples cultivated in different geographical areas. PMID- 11902921 TI - Selective responses of three Ginkgo biloba leaf-derived constituents on human intestinal bacteria. AB - The selective responses of Ginkgo biloba leaf-derived materials against six intestinal bacteria was examined using an impregnated paper disk method and compared with that of bilobalide, ginkgolides A and B, kaempferol, and quercetin. The components of G. biloba leaves were characterized as kaempferol 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside), kaempferol 3-O-(2' '-O-beta-D glucopyranosyl)-alpha-L-rhamnopyranoside, and quercetin 3-O-alpha-(6' "-p coumaroylglucosyl-beta-1,4-rhamnoside) by spectroscopic analysis. The growth responses varied with each bacterial strain tested. At 2 mg/disk, kaempferol 3-O alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) and quercetin 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) revealed potent inhibition against Clostridium perfringens, and kaempferol 3-O-(2' '-O-beta-D-glucopyranosyl)-alpha L-rhamnopyranoside showed a clear inhibitory effect on Escherichia coli. At 0.5 mg/disk, quercetin 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) showed a strong activity against C. perfringens, but weak activity was exhibited by kaempferol 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) against C. perfringens and kaempferol 3-O-(2' '-O-beta-D-glucopyranosyl)-alpha-L rhamnopyranoside against E. coli. No inhibition was observed from treatments conducted with bilobalide, ginkgolides A and B, kaempferol, or quercetin. Furthermore, these isolated compounds did not inhibit Bifidobacterium bifidum, B. longum, B. adolescentis, or Lactobacillus acidophilus. PMID- 11902922 TI - Antioxidant activity of chemical components from sage (Salvia officinalis L.) and thyme (Thymus vulgaris L.) measured by the oil stability index method. AB - A new abietane diterpenoid, 12-O-methyl carnosol (2), was isolated from the leaves of sage (Salvia officinalis L.), together with 11 abietane diterpenoids, 3 apianane terpenoids, 1 anthraquinone, and 8 flavonoids. Antioxidant activity of these compounds along with 4 flavonoids isolated from thyme (Thymus vulgaris L.) was evaluated by the oil stability index method using a model substrate oil including methyl linoleate in silicone oil at 90 degrees C. Carnosol, rosmanol, epirosmanol, isorosmanol, galdosol, and carnosic acid exhibited remarkably strong activity, which was comparable to that of alpha-tocopherol. The activity of miltirone, atuntzensin A, luteolin, 7-O-methyl luteolin, and eupafolin was comparable to that of butylated hydroxytoluene. The activity of these compounds was mainly due to the presence of ortho-dihydroxy groups. The 1,1-diphenyl-2 picrylhydrazyl radical scavenging activity of these compounds showed the similar result. PMID- 11902919 TI - Rapid fluorescence polarization immunoassay for the mycotoxin deoxynivalenol in wheat. AB - The fungus Fusarium graminearum, a pathogen of both wheat and maize, produces a toxin, deoxynivalenol (DON), that causes disease in livestock. A rapid test for DON in wheat was developed using the principle of fluorescence polarization (FP) immunoassay. The assay was based on the competition between DON and a novel DON fluorescein tracer (DON-FL2) for a DON-specific monoclonal antibody in solution. The method, which is a substantial improvement over our previous DON FP immunoassay, combined a rapid (3 min) extraction step with a rapid (2 min) detection step. A series of naturally contaminated wheat and maize samples were analyzed by both FP immunoassay and liquid chromatography (HPLC-UV). For wheat the HPLC-UV and FP methods agreed well (linear regression r(2) = 0.936), but for maize the two methods did not (r (2) = 0.849). We conclude that the FP method is useful for screening wheat, but not maize, for DON. PMID- 11902923 TI - Atmospheric pressure ionization LC-MS-MS determination of urushiol congeners. AB - This paper describes atmospheric pressure ionization (API) LC-MS-MS determination of urushiols, 3-n-alkenyl- and -alkyl-substituted catechols responsible for poison oak dermatitis. Urushiol was isolated from Western poison oak according to the method of Elsohly et al. (1) (J. Nat. Prod. 1982, 45, 532-538)-the purified preparation contained C(17)- and C(15)-substituted urushiols with zero, one, two, and three double bonds as determined from GC-MS analysis of trimethylsilyl derivatives. Urushiol mixtures were separated on a C(18) reversed phase HPLC column with a methanol-water gradient with urushiols eluting in 100% methanol. Atmospheric pressure chemical ionization (APCI) produced primarily [M - H](-) and MH(+) molecule ions. Electrospray ionization (ESI) yielded [M - H](-) and adduct ions including [M + Cl](-). Daughter ions of [M - H](-) included quinoid radical anions ([M - H - H(2)](-) and m/z 122(-)) and a benzofuran phenate (m/z 135(-)). A suite of hydrocarbon fragments were produced by collision-induced dissociation of MH(+) directly or via an intermediate [MH - H(2)O](+) daughter ion. Six urushiol congeners, one not previously reported in poison oak, were determined by negative ion API-LC-MS-MS with detection limits of approximately 8 pg/microL (ESI) and approximately 800 pg/microL (APCI). API-LC-MS-MS was used to determine urushiol in surface wipes, air samples, and plant materials. PMID- 11902924 TI - Profiling and characterization antioxidant activities in Anoectochilus formosanus hayata. AB - Phytochemical characteristics and antioxidant activities of the crude and fractionated plant extracts of Anoectochilus formosanus were evaluated using five different assay systems. An acid-treatment (2 N HCl in 95% ethanol) was employed to treat a butanol fraction (BuOH), creating an acid-hydrolyzed BuOH fraction. The IC(50) values for DPPH radicals in the BuOH and acid-hydrolyzed BuOH fractions were 0.521 and 0.021 mg/mL, respectively. The acid-hydrolyzed BuOH exhibited approximately 5-fold higher activity in scavenging superoxide anion than catechin. The acid-hydrolyzed BuOH fraction also effectively protected phi x174 supercoiled DNA against strand cleavage induced by H(2)O(2) and reduced oxidative stress in HL-60 cells. Metabolite profiling showed that the aglycones of flavonoid glycosides in BuOH were produced after acid hydrolytic treatment, and this resulted in a significant increase in antioxidant activities of acid hydrolyzed BuOH. One new diarylpentanoid, kinsenone, and three known flavonoid glycosides and their derivatives were identified for the first time from A. formosanus, with strong antioxidant properties. PMID- 11902925 TI - Larvicidal activity of isobutylamides identified in Piper nigrum fruits against three mosquito species. AB - The insecticidal activity of materials derived from the fruits of Piper nigrum against third instar larvae of Culex pipiens pallens, Aedes aegypti, and A. togoi was examined and compared with that of commercially available piperine, a known insecticidal compound from Piper species. The biologically active constituents of P. nigrum fruits were characterized as the isobutylamide alkaloids pellitorine, guineensine, pipercide, and retrofractamide A by spectroscopic analysis. Retrofractamide A was isolated from P. nigrum fruits as a new insecticidal principle. On the basis of 48-h LC(50) values, the compound most toxic to C. pipiens pallens larvae was pipercide (0.004 ppm) followed by retrofractamide A (0.028 ppm), guineensine (0.17 ppm), and pellitorine (0.86 ppm). Piperine (3.21 ppm) was least toxic. Against A. aegypti larvae, larvicidal activity was more pronounced in retrofractamide A (0.039 ppm) than in pipercide (0.1 ppm), guineensine (0.89 ppm), and pellitorine (0.92 ppm). Piperine (5.1 ppm) was relatively ineffective. Against A. togoi larvae, retrofractamide A (0.01 ppm) was much more effective, compared with pipercide (0.26 ppm), pellitorine (0.71 ppm), and guineensine (0.75 ppm). Again, very low activity was observed with piperine (4.6 ppm). Structure-activity relationships indicate that the N-isobutylamine moiety might play a crucial role in the larvicidal activity, but the methylenedioxyphenyl moiety does not appear essential for toxicity. Naturally occurring Piper fruit-derived compounds merit further study as potential mosquito larval control agents or as lead compounds. PMID- 11902926 TI - Authentication of Panax ginseng and Panax quinquefolius using amplified fragment length polymorphism (AFLP) and directed amplification of minisatellite region DNA (DAMD). AB - AFLP profiles characteristic to Panax ginseng and Panax quinquefolius were generated using primers E-AGG/M-CAA. P. ginseng samples from different farms in China and Korea are homogeneous genetically [similarity index (SI) = 0.88-0.99], whereas samples of P. quinquefolius from different sources are much more heterogeneous (SI = 0.64-0.96). Detailed analysis of one of the polymorphic bands in P. ginseng led to the identification of a minisatellite Pg2, which contains eight repeats of 5'-AGGACTCATCACATTGTTACTC. The minisatellite DNA was consequently used in directed amplification minisatellite region DNA analysis to authenticate the two ginsengs. PMID- 11902927 TI - Controlling molecular weight and degree of deacetylation of chitosan by response surface methodology. AB - Response surface methodology (RSM) was used for controlling molecular weight (MW) and degree of deacetylation (DOD) of chitosan in chemical processing. In a reduced model, MW of chitosan is y = 1736166.406 - 250.745X(1)X(2) - 265.452X(2)X(3), with R( 2) = 0.86, and DOD of chitosan is y = 30.6069 + 0.3396X(1) + 0.4948X(2) + 0.0094X(3)(2), with R( 2) = 0.89. MW of chitosan depends on the crossproduct of temperature and NaOH concentration and the crossproduct of NaOH concentration and time, and DOD depends linearly on temperature and NaOH concentration, and quadratically on time. Chitosan was widely depolymerized in a range from 1,100 kDa to 100 kDa and deacetylated from 67.3 to 95.7% by NaOH alkaline treatment. MW and DOD of chitosan were drastically decreased and increased, respectively, with increase of temperature, reaction time, and NaOH concentration. Furthermore, the rate of MW decrease and DOD increase of chitosan gradually decreased with prolonged reaction time. PMID- 11902928 TI - Flavor and texture of banana chips dried by combinations of hot air, vacuum, and microwave processing. AB - The behavior of 16 volatile compounds of banana during a combination of air drying (AD) and vacuum microwave-drying (VMD) of banana chips was characterized. Samples were AD to remove 60, 70, 80, or 90% of moisture (wet basis) and then subjected to VMD to achieve a final moisture content of 3% (dry basis). Banana slices were also dehydrated using only AD, VMD, and freeze-drying (FD) for comparison. Samples that underwent more VMD had significantly lower levels of volatile compounds, which is attributed to the decreased formation of an impermeable solute layer on the surface of the chips. High values for water solubility and relative volatility of compounds correlated with losses during VMD; however, additional factors appear to influence the behavior of compounds during VMD processing. The optimal process of 90%AD/10%VMD yielded crisper banana chips with significantly higher volatile levels and sensory ratings than AD chips. PMID- 11902929 TI - Effects of commercial processing on levels of antioxidants in oats (Avena sativa L.). AB - The effects of various commercial hydrothermal processes (steaming, autoclaving, and drum drying) on levels of selected oat antioxidants were investigated. Steaming and flaking of dehulled oat groats resulted in moderate losses of tocotrienols, caffeic acid, and the avenanthramide Bp (N-(4'-hydroxy)-(E) cinnamoyl-5-hydroxy-anthranilic acid), while ferulic acid and vanillin increased. The tocopherols and the avenanthramides Bc (N-(3',4'-dihydroxy-(E)-cinnamoyl-5 hydroxy-anthranilic acid) and Bf (N-(4'-hydroxy-3'-methoxy)-(E)-cinnamoyl-5 hydroxy-anthranilic acid) were not affected by steaming. Autoclaving of grains (including the hulls) caused increased levels of all tocopherols and tocotrienols analyzed except beta-tocotrienol, which was not affected. Vanillin and ferulic and p-coumaric acids also increased, whereas the avenanthramides decreased, and caffeic acid was almost completely eliminated. Drum drying of steamed rolled oats resulted in an almost complete loss of tocopherols and tocotrienols, as well as a large decrease in total cinnamic acids and avenanthramides. The same process applied to wholemeal made from groats from autoclaved grains resulted in less pronounced losses, especially for the avenanthramides which were not significantly affected. PMID- 11902930 TI - Changes in the cell wall network during the thermal dehydration of alfalfa stems. AB - The effects of heat treatments used to dry alfalfa stems were investigated. Heating at 70 or 100 degrees C caused no major change in the cell wall composition, but xylanase had lower activity on the cell wall of heated material and the amount of xylose released varied with the temperature used. Chemical fractionation of cell wall carbohydrates showed that the main changes occurring during stem dehydration concerned pectic polymers and probably hemicelluloses. There was less material soluble in ammonium oxalate from alfalfa heated at 100 degrees C than from fresh alfalfa. The results suggest that heat processing causes some changes in the cell wall network. Environmental scanning electron microscopy was used to examine fully hydrated tissues at high resolution. There was cell distortion without disruption of cell walls as water was lost. PMID- 11902931 TI - Flavonoid and carbohydrate contents in Tropea red onions: effects of homelike peeling and storage. AB - The content of anthocyanins, flavonols, and carbohydrates of Tropea red onions (Allium cepa L.) was determined by HPLC and HPLC-MS. Cyanidin derivatives constitute >50% of total anthocyanins, but delphinidin and petunidin derivatives, which have not been reported in red onions thus far, were also detected. The flavonoid distribution in the different layers of the bulbs indicates that, after homelike peeling, the edible portion contains 79% of the total content of quercetin 4'-glucoside but only 27% of the anthocyanins. Storage of onions for 6 weeks in different conditions, all of them mimicking home storage habits, resulted in a decrease to 64-73% of total anthocyanins. The same trend was verified for the total antioxidant activity, which was reduced to 29-36%. A decrease in glucose and fructose content correlated with anthocyanin degradation was also observed. Storage at low temperature seems to better preserve the onion anthocyanins. PMID- 11902932 TI - SAR studies of sesquiterpene lactones as Orobanche cumana seed germination stimulants. AB - Studies of the structure-activity relationship (SAR) directed to evaluate the effect of several sesquiterpene lactones (SL) as germination stimulants of three Orobanche spp. (O. cumana, O. crenata, and O. ramosa) have been achieved. Results are compared with those obtained in the same bioassay with an internal standard, the synthetic analogue of strigol GR-24. A high specificity in the germination activity of SL on the sunflower parasite O. cumana has been observed, and a relationship between such activity and the high sunflower SL content is postulated. Molecular properties of the natural and synthetic germination stimulants (GR-24, GR-7, and Nijmegen-1) and SL have been studied using MMX and PM3 calculations. Consequently, comparative studies among all of them and their activities have been made. SL tested present similarities in molecular properties such as the volume of the molecule and the spatial disposition of the carbon backbone to the natural germination stimulant orobanchol. These properties could be related to their biological activity. PMID- 11902933 TI - FTIR study of glyphosate-copper complexes. AB - Complexes of the herbicide glyphosate (GPS) and the heavy metal Cu were studied by infrared spectroscopy under controlled pH, in order to know the mechanisms involved in the formation of these complexes. In CuGPS(-), the IR spectrum shows participation of the carboxylate and phosphonic moieties of the GPS molecule. The formation of the complex produces a lower symmetry in the phosphonate group because of loss of the resonance situation of PO(3)(2)(-) groups, with a subsequent split of their absorption bands. Carboxylate groups are participating by forming unidentate complexes. No conclusion is reached about the involvement of the amino group, but previous EPR findings indicate coordination of GPS to Cu via nitrogen. Consequently, glyphosate in this complex functions with a tridentate character by forming two chelate rings. A study of the CuGPSH species was not possible due to overlapping of its absorption bands with those of free GPS species. PMID- 11902934 TI - Identification of fonofos metabolites in Latuca sativa, Beta vulgaris, and Triticum aestivum by packed capillary flow fast atom bombardment tandem mass spectrometry. AB - The metabolism of fonofos, a thiophosphonate insecticide, was investigated in mature lettuce (Latuca sativa), beet (Beta vulgaris), and wheat (Triticum aestivum). Six new metabolites were identified by LC-MS and LC-MS-MS analysis using fast atom bombardment (FAB) and packed capillary LC columns with application of the on-column focusing technique. These methods provided the sensitivity required to identify unknown metabolites that were present in the mature plants at only 20-230 ppb. Structural elucidation was facilitated by use of fonofos labeled with both carbon-14 and carbon-13 in the phenyl ring. In all three plants fonofos was converted to a glucose conjugate of thiophenoxylactic acid. Oxidation of the glucose conjugate gave isomeric sulfoxides in all species examined. Thiophenoxylactic acid was found esterified to malonic acid in lettuce. In beets, S-phenylcysteine was found as its malonic acid amide. A second metabolite unique to beets was N-(malonyl) [2[(ethoxyethylphosphinothionyl)oxy]phenyl]cysteine. This novel structure was confirmed by synthesis. PMID- 11902935 TI - Disappearance of azoxystrobin, pyrimethanil, cyprodinil, and fludioxonil on tomatoes in a greenhouse. AB - The disappearance of azoxystrobin, pyrimethanil, cyprodinil, and fludioxonil on tomatoes in greenhouse was studied. At the preharvest interval, except for cyprodinil, the pesticide residues were below the MRL fixed in Italy. The mechanism of disappearance studied with model systems shows that the decrease in residues was due to codistillation and photodegradation in pyrimethanil, to photodegradation in fludioxonil, and to evaporation and codistillation in cyprodinil. Azoxystrobin residues were stable during all experiments. PMID- 11902936 TI - Soybean (Glycine max) cell wall composition and availability to feed enzymes. AB - Defatted untoasted soybean cotyledons and hulls were fractionated as water solutes (WSc and WSh) and water unextractable (WUc and WUh). Further fractionation of WUc through deproteinization yielded the isolation of a water unextractable solid (WUS) fraction that was mainly composed (molar percent) of galactose (28.1%), glucose (27.8%), arabinose (13.3%), and uronic acids (17.6%), which accounted for 76% of the water insoluble polysaccharides in soybean cotyledons (WUc). The cell wall (WUS) was sequentially fractionated with chelating agents (chelating agent soluble solids, ChSS) and a gradient of agents (dilute alkali, DASS; 1 M alkali, 1MASS; and 4M alkali, 4MASS), which gave a final cellulosic residue. The ChSS and DASS extracts were characterized as pectin rich fractions, whereas 1MASS and 4MASS were hemicellulose- and cellulose-rich fractions. Incubation in vitro of the WUc fraction with pectinase, cellulase, and xylanase resulted in the release of low amounts (not more than 5% bound basis) of monosaccharides, mostly uronic acids, xylose, and arabinose. Protein extraction hardly increased this release after enzymatic incubation (<7%). However, progressive fractionation of the cell wall matrix markedly increased the release of monosaccharides from pectin- (ChSS and DASS) and hemicellulose-rich fractions (1MASS and 4MASS). Significant degradation of cellulose (up to 20%) was achieved only after complete protein, pectin, and hemicellulose extraction. PMID- 11902937 TI - Characterization of the major proteins of tubers of yam bean (Pachyrhizus ahipa). AB - Tubers of six accessions of ahipa (Pachyrhizus ahipa) contained between 0.77 and 1.34% nitrogen on a dry weight basis. This corresponds to 4.8 to 8.4% crude protein based on a nitrogen to protein conversion factor of 6.25; but detailed analysis of AC230 showed that although 93% of the total N was extracted with buffer containing 1.0 M NaCl, about a third of this was lost on dialysis. It was calculated, therefore, that salt-soluble proteins comprise about 60% of the total tuber nitrogen, with low-molecular-mass nitrogenous components comprising a further 30%. Electophoretic analysis of the salt-soluble proteins showed similar patterns of components in the six accessions, with none being present in amounts sufficiently high to suggest a role as storage proteins. Furthermore, light microscopy failed to show significant deposits of protein within the tuber cells. Five "major" protein bands, which together accounted for about 19% of the total salt-soluble protein fraction were purified and subjected to N-terminal amino acid sequencing. Comparison of these with sequences in protein databases revealed similarities to alpha-amylases, chitinases and chitin binding proteins, cysteine proteinases (including major components from P. erosus tubers), a tuberization specific protein from potato, and proteins induced in soybean and pea by stress or the plant hormone abscisic acid, respectively. It was concluded that the primary roles of these proteins are probably in aspects of tuber metabolism and development and/or conferring protection to pests and pathogens, and that true storage proteins are not present. The absence of storage proteins is consistent with the biological role of the tubers as storage organs for carbohydrates (cf cassava tuberous roots) rather than as propagules (cf yam and potato tubers). PMID- 11902938 TI - Analysis of products of animal origin in feeds by determination of carnosine and related dipeptides by high-performance liquid chromatography. AB - Products of animal origin such as meat meal were commonly used as sources of protein and amino acids for the production of compound feeds. Because the feeding of such products is prohibited in Germany, the official feedstuff control of the government must evaluate feeds for the forbidden use of products of animal origin. Microscope examination is the official method to prove animal-originated adulterations of feeds. This paper proposes a high-performance liquid chromatography method for the determination of the dipeptide carnosine and related dipeptides (anserine and balenine) and shows the dependence of the contents of anserine, balenine, and carnosine in compound feeds on the content of meat meal in feeds. The presented method can complete and confirm the result of the microscopic method for evidence of components of animal origin in feeds. PMID- 11902939 TI - Enzymatic cleaning of inorganic ultrafiltration membranes fouled by whey proteins. AB - The aim of this work was to study the cleaning of inorganic membranes fouled with whey protein solutions using the enzymatic formulation Alcalase (Novo Nordisk A/S). Hydraulic and chemical methods were considered to characterize the cleanliness of the membranes. Cleaning efficiency was observed to be a function of the operating conditions. The operating conditions tested were the following: recycling versus non-recycling of permeate, pH of the cleaning solution, addition of alkali to regulate the pH, enzymatic agent concentration, and cleaning time. The best conditions to perform the cleaning were related to the best conditions to hydrolyze whey proteins in a discontinuous reactor using the same enzyme preparations. Very high cleaning efficiencies (>90%) were achieved in short operating times (20 min). However, residual matter was observed on the membrane surface. PMID- 11902940 TI - Dissipation of monosodium methane arsonate (MSMA) on peanuts. AB - Monosodium methane arsonate (MSMA), the salt of methylarsonic acid (MAA), is a herbicide commonly used to control weeds along roadsides, in cotton, in turf, and on noncrop sites. In recent years questions have arisen regarding the source and nature of arsenic residues in raw agricultural commodities relative to misuse, inadvertent exposure, or rotational crop residues. Field experiments were conducted to determine the fate of MSMA that is applied to peanut foliage. Persistence, dissipation, recovery, and detection of MAA from leaf rinsate were characterized as well as resulting total arsenic and MAA concentrations in peanut kernels. MSMA was applied to peanut foliage at 105, 210, 315, and 420 g of active ingredient (ai)/ha. Peanut leaves were sampled before and after irrigation events over the next 7 days. Peanuts were harvested at maturity and analyzed for MAA and total arsenic. A confined rotational crop experiment was conducted to determine the potential for MAA residues in soil to be taken up by peanuts in fields rotated from cotton that was treated with MSMA. MAA was not detected in any peanut samples from the rotational crop experiment, even when peanuts were planted only 30 days after MSMA application to the soil at 2.24 kg of ai/ha. Field experiments showed that MSMA recovery from leaves with an aqueous rinse declined quickly but was not greatly affected by irrigation. However, quantifiable amounts of MAA were present 1 week after application and after two irrigation events, and MAA and total arsenic were measured in mature peanut kernels from all plots that received MSMA. MAA was not detected in untreated checks. Total arsenic was below the limit of quantification in untreated controls. PMID- 11902941 TI - Accumulation of Zn, Cd, Cu, and Pb in Chinese cabbage as influenced by climatic conditions under protected cultivation. AB - Accumulation of heavy metals from agricultural soils contaminated by low levels heavy metals has important implications in the understanding of heavy metal contamination in the food chain. Through field experiments (1994-1996), the influence of thermal regime under different treatments on the accumulation of zinc, cadmium, copper, and lead in Chinese cabbage [Brassica pekinensis (Lour) Rupr. cv. Nagaoka 50] grown in a Calcareous Fluvisol (Xerofluvent) in Granada (southern Spain) was examined. Two floating row covers were used: T(1) (perforated polyethylene, 50 microm thick) and T(2) (17 g m(-2) polypropylene nonwoven fleece). An uncovered cultivation (T(0)) served as control. Zn, Cd, Cu, and Pb levels in the whole tops of experimental plants were analyzed. Treatments T(1) and T(2) gave rise to differences in environmental conditions with respect to T(0). The influence of environmental factors manipulated by floating row covers (particularly under T(1)) increased total heavy metal accumulation in the above ground plant biomass with respect to the open-air crop. The total contents of Zn, Cd, Cu, and Pb were 30, 50, 90, and 40% higher in T(1), respectively, than in T(0). This technique could be used in contaminated zones for different plant species because the thermal effect favors the process of phytoextraction and thus reduces the contamination. PMID- 11902942 TI - Determination of semivolatile compounds in Baltic herring (Clupea harengus membras) by supercritical fluid extraction-supercritical fluid chromatography-gas chromatography-mass spectrometry. AB - The on-line supercritical fluid extraction-supercritical fluid chromatography-gas chromatography method was applied to the determination of volatile compounds of raw and baked Baltic herring (Clupea harengus membras). The analytes were extracted with supercritical carbon dioxide at 45 degrees C and 10 MPa pressure. After extraction, the volatiles and coeluted lipids were separated on-line using supercritical fluid chromatography and the volatile fraction was introduced directly into a gas chromatograph. In all, 30 compounds were identified from fish samples with mass spectrometry. The most abundant compounds in the fresh Baltic herrings were heptadecane and 1-heptadecene. When the fish were stored for 3-6 days at 6 degrees C, the total peak area of the volatiles began to increase and the proportions of short chain acids (acetic, propanoic, 2-methylpropanoic, and 3 methylbutanoic) also increased. After 8-9 days of storage, 3-methylbutanoic acid comprised about 36 and 40% of all volatiles in raw and baked herring, respectively. PMID- 11902943 TI - Effect of gelatinization and starch-emulsifier interactions on aroma release from starch-rich model systems. AB - Release of selected volatile aldehyde compounds from starch-rich matrices was studied by headspace extraction, using solid-phase microextraction, and gas chromatography. Changes in the rheological properties of the starch-based matrices, due to starch concentration, gelatinization, and interactions with emulsifiers, were studied by steady shear and dynamic methods. The degree of volatile retention was found to depend on the compound properties, starch concentration, native structure of the granules, and presence of emulsifiers. The nongelatinized starch granules were more effective in lowering the volatile headspace quantities. Loss of the native structural integrity of the granules decreases the retention ability. For the nongelatinized starch dispersions, the more hydrophilic emulsifier showed a more pronounced effect on the matrix rheology and also on the aroma retention. A different behavior was observed for the gelatinized systems. Interpretation of the volatile release profiles was made on the basis of the matrix physical properties and interactions among components. PMID- 11902944 TI - Modeling the partition of volatile aroma compounds from a cloud emulsion. AB - Parameters determining the partitioning behavior of volatile compounds between a cloud emulsion and the gas phase were measured under static equilibrium headspace conditions, using volatiles (e.g., ethyl hexanoate, cymene, and octanol) representing different volatilities and different degrees of hydrophobicity. The significant factors were the molecular characteristics of the volatile and the concentration of the oil phase. The nature of the lipid (C8 and C12 triglycerides), particle size, and emulsifier type (modified starch and gum arabic) did not significantly alter volatile partitioning. An empirical model based on the partition behavior and physicochemical parameters of 67 volatile compounds was produced. This predicted the partition of volatiles (R(2) = 0.83) in cloud emulsions as a function of lipid content. The significant terms (P < 0.05) in the empirical model were Log P, Log solubility, the dipole vector, and the oil fraction. PMID- 11902945 TI - Purification of hydroperoxide lyase from green bell pepper (Capsicum annuum L.) fruits for the generation of C6-aldehydes in vitro. AB - The aim of this work was to compare the efficiency of different extracts of hydroperoxide lyase from green bell peppers in producing aldehydes: a crude extract, a chloroplastic fraction, and a purified enzyme were investigated. From a crude extract, the HPO lyase was purified by ion-exchange chromatography with a 22.3-fold increase in purification factor. Analysis by SDS-PAGE electrophoresis under denaturating conditions showed only one protein with a molecular weight of 55 kDa, whereas size-exclusion chromatography indicated a molecular weight of 170 kDa. A maximum of 7500 mg of aldehydes per g of protein was obtained with the purified enzyme within 20 min of bioconversion compared to 392 and 88 mg of aldehydes per g of protein within 50 and 60 min, respectively, for the chloroplast fraction and the crude extract. PMID- 11902946 TI - Characterization of the most odor-active compounds of Iberian ham headspace. AB - Gas chromatography-olfactometry (GC-O) based on detection frequency (DF) was used to characterize the most odor-active compounds from the headspace of Iberian ham. Twenty-eight odorants were identified by GC-O on two capillary columns, including aldehydes (11), sulfur-containing compounds (7), ketones (5), nitrogen-containing compounds (2), esters (2), and an alcohol. Among them, the highest odor potencies (DF values) were found for 2-methyl-3-furanthiol, 2-heptanone, 3-methylbutanal, methanethiol, hexanal, hydrogen sulfide, 1-penten-3-one, 2-methylpropanal, ethyl 2-methylbutyrate, and (E)-2-hexenal. Nine of the 28 most odor-active compounds were identified for the first time as aroma components of dry-cured ham, including hydrogen sulfide, 1-penten-3-one, (Z)-3-hexenal, 1-octen-3-one, and the meaty-smelling compounds 2-methyl-3-furanthiol, 2-furfurylthiol, 3-mercapto-2 pentanone, 2-acetyl-1-pyrroline, and 2-propionyl-1-pyrroline. PMID- 11902947 TI - Precursors of 2-acetyl-1-pyrroline, a potent flavor compound of an aromatic rice variety. AB - The biological formation of a potent flavor compound, 2-acetyl-1-pyrroline, in the aromatic rice variety (Khao Dawk Mali 105) was studied in seedlings and callus of the rice. Concentrations of 2-acetyl-1-pyrroline were determined by GC MS-SIM using an isotope dilution method. Increases in concentration occurred when proline, ornithine, and glutamate were present in solution, with proline increasing the concentration by more than 3-fold compared to that of the control. Results of tracer experiments using (15)N-proline, (15)N-glycine, and proline-1 (13)C indicated that the nitrogen source of 2-acetyl-1-pyrroline was proline, whereas the carbon source of the acetyl group was not the carboxyl group of proline. 2-acetyl-1-pyrroline was formed in the aromatic rice at temperatures below that of thermal generation in bread baking, and formed in the aerial part of aromatic rice from proline as the nitrogen precursor. PMID- 11902948 TI - Identification of malodorous, a wild species allele affecting tomato aroma that was aelected against during domestication. AB - Vegetable cultivation favored the inclusion of pleasant aromas in the produce, whereas unpleasant aromas were selected against. Introgression lines, generated by hybridization of a cultivated tomato (Lycopersicon esculentum) to its wild relative L. pennellii, were used to map quantitative trait loci (QTL) that influence tomato aroma. A marked undesirable flavor was detected by taste panelists in L. pennellii fruits and was related to an introgressed segment from the short arm of chromosome 8. Analysis of the ripe fruits' volatiles of chromosome 8 introgressed lines revealed an up to 60-fold increase in the levels of 2-phenylethanol and phenylacetaldehyde, as compared to the cultivated tomato. This effect was associated with a 10 cM segment originating from the wild species. Although 2-phenylethanol and phenylacetaldehyde have favorable contribution to tomato aroma when present at low levels, phenylacetaldehyde has a nauseating objectionable aroma when present in levels >0.005 ppm. The loss of the ability to produce high levels of phenylacetaldehyde contributed to the development of desirable aroma of the cultivated tomato. The findings provide a genetic explanation for one of the aroma changes that occurred during the domestication of the tomato. PMID- 11902949 TI - Comparison of volatile compounds in water- and oil-soluble annatto (Bixa orellana L.) extracts. AB - Annatto is a natural food colorant extracted from the seeds of the Bixa orellana L. plant. Annatto is used in Latin American cuisine to add a deep red color as well as distinctive flavor notes to fish, meat, and rice dishes. In the United States, annatto extracts are primarily used to impart orange/yellow hues to cheese and other dairy foods. The objective of this study was to identify and compare volatile compounds present in water- and oil-soluble annatto extracts. Volatile compounds were recovered using dynamic headspace-solvent desorption sampling and analyzed using GC-MS. Compounds were identified by comparison to a mass spectral database, Kovats indexes, and retention times of known standards. Of the 107 compounds detected, 56 compounds were tentatively identified and 51 were positively identified. Volatile profile differences exist between water- and oil- soluble extracts, and annatto extracts contain odorants with the potential to influence food aroma. PMID- 11902951 TI - Accelerated shelf life testing of whey-protein-coated peanuts analyzed by static headspace gas chromatography. AB - Four different formulations of whey-protein-based coatings were used to coat peanuts. Four controls were used to investigate the effects of different ingredients in the coating formulation on the peanut shelf life. Untreated peanuts were designated as the reference. The peanut samples were stored in duplicate at 40, 50, and 60 degrees C for storage durations of up to 31 weeks. The analysis of hexanal indicated that the coated samples were oxidized significantly slower than the reference; hence, the predicted shelf life was longer for the coated samples. However, the investigation of the control ingredients revealed that even when only water was applied onto the peanuts the oxidation was delayed. PMID- 11902950 TI - Identification and quantification of aroma-active components that contribute to the distinct malty flavor of buckwheat honey. AB - Characteristic aroma components of buckwheat honey were studied by combined sensory and instrumental techniques. Relative aroma intensity of individual volatile components was evaluated by aroma extract dilution analysis (AEDA) of solvent extracts and by gas chromatography-olfactometry (GCO) of decreasing headspace samples (GCO-H). Results indicated that 3-methylbutanal, 3-hydroxy-4,5 dimethyl-2(5H)-furanone (sotolon), and (E)-beta-damascenone were the most potent odorants in buckwheat honey, with 3-methylbutanal being primarily responsible for the distinct malty aroma. Other important aroma-active compounds included methylpropanal, 2,3-butanedione, phenylacetaldehyde, 3-methylbutyric acid, maltol, vanillin, methional, coumarin, and p-cresol. PMID- 11902952 TI - Metal ions restore the proteolytic resistance of denatured conglutin gamma, a lupin seed glycoprotein, by promoting its refolding. AB - The susceptibility to trypsin of conglutin gamma, a lupin seed glycoprotein affected by this enzyme only when in a non-native conformation, was used to study the effect of Zn(2+) and other metal ions on the structural dynamics of the protein. When acid-treated trypsin-susceptible conglutin gamma was incubated at neutral pH in the presence of Zn(2+), it became resistant to tryptic attack, contrary to the protein treated in the absence of Zn(2+). The time course of this refolding event has been quantitatively evaluated by SDS-PAGE. Amino acid sequencing of the major polypeptide fragments, produced by trypsin before completion of the refolding process, indicated that only a few cleavable bonds were accessible to the enzyme. This suggested that the presence of metal ions affected the pathway of degradation of the protein, by inducing its folding. Among the other metal ions tested, Ni(2+) also promoted the adoption of a trypsin resistant conformation of conglutin gamma, whereas Mn(2+) and Ca(2+) had only much lower effects. The relevance of these findings for a deeper understanding of the in vivo degradation of plant food proteins and how it is affected by metal ions are discussed. PMID- 11902953 TI - Isolation and analysis of kappa-casein glycomacropeptide from goat sweet whey. AB - Glycomacropeptide (GMP) was purified from goat sweet whey by anion-exchange and hydrophobic interaction chromatography. Approximately 0.06% (w/v) of sweet whey was recovered as GMP. Amino acid analysis of the GMP preparation showed that the content of phenylalanine (an amino acid that does not occur in goat GMP) was negligible, indicating that the GMP was of high purity. The goat GMP contained 25 microg sialic acid per mg of dry weight. This was approximately 3-fold lower than the sialic acid concentration in bovine GMP reported in the literature. Gel electrophoretic results demonstrated that most of the goat GMP occurs as a dimer. The GMP was intensely stained with Coomassie blue in 50% methanol containing 12.5% (w/v) trichloroacetic acid, but showed very weak metachromasia with the same dye in 45% methanol containing 10% acetic acid, a preparation commonly used to stain protein. PMID- 11902954 TI - Comparative quality assessment of cultured and wild sea bream (Sparus aurata) stored in ice. AB - Comparative quality assessment of cultured and wild sea bream stored in ice for up to 23 days was achieved by the monitoring of sensory quality, levels of nucleotide, nucleotide breakdown products, and texture by a texturometer. The changes in sensory quality of both raw and cooked fish were assessed using the modified Tasmanian and Torry schemes, respectively. K and related values (freshness indicators), namely, K, K(i), G, P, H, and F(r), were calculated. Linear increases (r(2) > or = 0.99) in K, K(i), G, and P (and a decrease in F(r)) values for cultured sea bream and in the H value for wild sea bream with increasing storage periods were observed. The limit for acceptability of cultured and wild sea bream stored in ice was approximately 16-18 days (average K, K(i), G, and P values: approximately 35-40%; H values: approximately 5% for cultured and 10% for wild; and F(r ) values: approximately 65-70%). The texture of cultured and wild sea bream decreased throughout the storage period, and they were not significantly (p > 0.05) different until after day 16 when the wild sea bream was significantly softer than the cultured. The sensory score of both cultured and wild raw fish showed a good relationship with some freshness and texture indicators over the entire storage period (r(2) values > or = 0.99). These indicators were K, K(i), G, P, and F(r) values for cultured and H value for wild fish. PMID- 11902955 TI - Levels of stilbene oligomers and astilbin in French varietal wines and in grapes during noble rot development. AB - Phenolics from grapes and wines can play a role against oxidation and development of atherosclerosis. Stilbenes have been shown to have cancer chemopreventive activity and to protect lipoproteins from oxidative damage. A method for the direct determination of stilbene oligomers (viniferin and pallidol) as well as astilbin in different types of wine using high-performance liquid chromatography with UV detection is described. In a survey of 21 commercial wines from the south of France, levels of pallidol and viniferin are reported for the first time in different types of wines. Viniferin was found to be present only in red and botrytized sweet white wines with levels between 0.1 and 1.63 mg/L; pallidol was not found in dry and sweet white wines but only in wines made by maceration with stems, with levels between 0.38 and 2.22 mg/L. Highest levels of astilbin were found in Egiodola (15.13 mg/L), Merlot (11.61 mg/L), and Cabernet Sauvignon (8.24 mg/L) for red wines and in Sauvignon (5.04 mg/L) for white varietal wines. Astilbin levels are highest for recent vintages, but pallidol is not found in older vintages. During noble rot development in Sauvignon or Semillon grapes from the Sauternes area, levels of trans-astringin, trans-resveratrol, trans-piceid, and pallidol are quite low (<0.5 mg/kg of grapes). Viniferin and astilbin levels become optimum at 2 and 30 mg/kg, respectively, during spot grape and speckle grape stages. PMID- 11902956 TI - Effect of antioxidants, citrate, and cryoprotectants on protein denaturation and texture of frozen cod (Gadus morhua). AB - To investigate the role of antioxidants and cryoprotectants in minimizing protein denaturation in frozen lean fish, cod fillets were treated with either antioxidants (vitamin C (500 mg kg(-1)) or vitamin C (250 mg kg(-1)) + vitamin E (250 mg kg(-1))), antioxidants (vitamins C + E 250 mg kg(-1)each) with citrate (100 mg kg(-1)), cryoprotectants (4% (w/w) sucrose + 4% (w/w) sorbitol), or a mixture of antioxidants (vitamins C + E 250 mg kg(1)), citrate (100 mg kg(-1)), and cryoprotectants (sucrose 40 g kg(-1) + sorbitol 40 g kg(-1)). Untreated and treated fish samples were stored at -10 degrees C; cod fillets stored at -30 degrees C were used as a control. Stored frozen samples were analyzed at intervals for up to 210 days for changes in protein extractability, thermodynamic parameters (transition temperature T(m) and enthalpy DeltaH), structure by FT Raman spectroscopy, and rheological properties by large and small deformation tests. Results indicated that protein denaturation and texture changes were minimized in the presence of cryoprotectants, as well as in the presence of antioxidants with citrate, antioxidants alone, or the mixture of antioxidants, citrate, and cryoprotectants. In the presence of increased formaldehyde levels in fish treated with vitamin C, toughening was still lower compared to that of the 10 degrees C control due to the antioxidant property of vitamin C. Thus, ice crystal formation and lipid oxidation products are the major factors that cause protein denaturation in lean frozen fish, and antioxidants in addition to cryoprotectants can be used to minimize toughness. PMID- 11902958 TI - Chemical profiling to differentiate geographic growing origins of coffee. AB - The objective of this research was to demonstrate the feasibility of this method to differentiate the geographical growing regions of coffee beans. Elemental analysis (K, Mg, Ca, Na, Al, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Mo, S, Cd, Pb, and P) of coffee bean samples was performed using ICPAES. There were 160 coffee samples analyzed from the three major coffee-growing regions: Indonesia, East Africa, and Central/South America. A computational evaluation of the data sets was carried out using statistical pattern recognition methods including principal component analysis, discriminant function analysis, and neural network modeling. This paper reports the development of a method combining elemental analysis and classification techniques that may be widely applied to the determination of the geographical origin of foods. PMID- 11902957 TI - Effect of caffeic acid on the color of red wine. AB - The copigmentation effect of prefermentation additions of different doses of caffeic acid was investigated during the 1997 harvest. Microfermentation with the major red grape cultivars Listan negro and Negramoll, grown in the Canary Islands, was carried out with the same protocol. Visible and UV spectra were registered periodically. HPLC chromatograms were carried out. The color enhancement of cv. Negramoll wine varied between 13 and 75% (AU at 520 nm), and that of cv. Listan negro wine between 25 and 45% at the end of fermentation. During aging these values were enhanced to reach even >100% in some cases. An initial complex of the 1:1 type, where one molecule of caffeic acid associates with one molecule of anthocyanin, has been identified using the mathematical procedure of Brouillard et al. (J. Am. Chem. Soc. 1989, 111, 2604-2610). Caffeic acid seems to contribute to color stability and protection against oxidation. The importance of nonpigment composition in pigment extraction and color retention during and after fermentation is demonstrated. PMID- 11902959 TI - Chemometric studies of chemical compounds in five cultivars of potatoes from Tenerife. AB - A statistical study of correlation and multivariate analysis on the chemical composition of five cultivars of potatoes harvested in Tenerife was carried out to establish the relationships between the chemical compounds and, therefore, to differentiate the samples according to traditional and recent importation potatoes, cultivars, and species/subspecies. A large number of significant correlations between the chemical compounds were found, which suggests biochemical relationships among them. After factor analysis, the dimension space was reduced from 24 variables to eight factors, accounting for 77.2% of the total variance. Starch, moisture, organic acids, and metals are the variables that make it possible to characterize the system without losing very much information. Total differentiation of potato samples according to the criteria species/subspecies and cultivars was obtained using discriminant analysis with all the variables. However, with only four variables (weight of tuber, starch, amylose, and glucose + fructose) it is possible to differentiate between the traditional and recent importation potatoes. PMID- 11902961 TI - Distribution of gamma-glutamyl-beta-alanylhistidine isopeptide in the macromolecular fractions of commercial meat extracts and correlation with the color of the macromolecular fractions. AB - The measurement of gamma-glutamyl-beta-alanylhistidine isopeptide in the macromolecular fraction of various commercial meat extracts indicated that all of the commercial meat extracts tested contained the isopeptide, in concentrations ranging from 0.04 to 0.87 micromol/g of dry matter. This variation was suggested to be due to the differences between the processes of extraction and the differences in the initial amounts of carnosine. A positive correlation between the content of gamma-glutamyl-beta-alanylhistidine and the color of the macromolecular fraction was observed. These results suggested that gamma-glutamyl beta-alanylhistidine is widely distributed in meat products and that the content can be used as an index of protein denaturation during the heating process. PMID- 11902960 TI - Intact carbohydrate structures as part of the melanoidin skeleton. AB - Model melanoidins from monomeric, oligomeric, and polymeric carbohydrates, and amino acids formed under aqueous as well as water-free reaction conditions, were submitted to acidic catalyzed hydrolysis. Their degradation products were detected qualitatively and quantitatively by HPTLC and HPLC-DAD. A considerable amount of monomer carbohydrates from hydrolysis of model melanoidins formed under water-free reaction conditions was detected. It can be seen clearly that the amount of carbohydrates released increased with increasing degree of polymerization of the carbohydrates used as starting material. In comparison, the hydrolysis of melanoidins formed in aqueous condition resulted in only a small glucose release. It seems that in the Maillard reaction under water-free conditions, a significant amount of di- and oligomer carbohydrates were incorporated into the melanoidin skeleton as complete oligomer with intact glycosidic bond, forming side chains at the melanoidin skeleton. Additional side chains could be formed by transglycosylation reactions. With increasing water content, hydrothermolytic as well as retro-aldol reactions of the starting carbonyl components became significant, and therefore the possibility of forming side chains decreased. The results are consistent with the postulated melanoidin structure being built up mainly from sugar degradation products, probably branched via amino compounds. PMID- 11902962 TI - Oxidative stability of fish and algae oils containing long-chain polyunsaturated fatty acids in bulk and in oil-in-water emulsions. AB - The oxidative stability of long-chain polyunsaturated fatty acid (PUFA) and docosahexaenoic acid (DHA)-containing fish and algae oils varies widely according to their fatty acid composition, the physical and colloidal states of the lipids, the contents of tocopherols and other antioxidants, and the presence and activity of transition metals. Fish and algal oils were initially much more stable to oxidation in bulk systems than in the corresponding oil-in-water emulsions. The oxidative stability of emulsions cannot, therefore, be predicted on the basis of stability data obtained with bulk long-chain PUFA-containing fish oils and DHA containing algal oils. The relatively high oxidative stability of an algal oil containing 42% DHA was completely lost after chromatographic purification to remove tocopherols and other antioxidants. Therefore, this evidence does not support the claim that DHA-rich oils from algae are unusually stable to oxidation. Addition of ethylenediaminetetraacetic acid (EDTA) prevented oxidation of both fish and algal oil emulsions without added iron and at low iron:EDTA molar concentrations. EDTA, however, promoted the oxidation of the corresponding emulsions that contained high iron:EDTA ratios. Therefore, to be effective as a metal chelator, EDTA must be added at molar concentrations higher than that of iron to inhibit oxidation of foods containing long-chain PUFA from either fish or algae and fortified with iron. PMID- 11902963 TI - Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays). AB - Polymerase Chain Reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. In this paper, recombinant DNAs introduced into the seven lines of GM maize, such as Event 176, Bt11, T25, MON810, GA21, DLL25, and MON802, are sequenced. On the basis of the obtained sequence, 14 primer pairs for the detection of the segments, such as promoter, terminator regions, and construct genes, were designed. To confirm the specificities of the designed primer pairs, PCR was performed on genomic DNAs extracted from GM and non-GM maize, GM and non-GM soy, and other cereal crops. Because the presence of the corresponding DNA segments was specifically detected in GM crops by the designed primer pairs, it was concluded that this method is useful for fast and easy screening of GM crops including unauthorized ones. PMID- 11902964 TI - Identification of anthocyanin-flavanol pigments in red wines by NMR and mass spectrometry. AB - Three newly formed Port wine pigments were isolated by Toyopearl HW-40(s) gel chromatography and semipreparative HPLC. Furthermore, the pigments were identified by mass spectrometry (LC/MS) and NMR techniques (1D and 2D). These anthocyanin-derived pigments showed UV-visible spectra different from those of the original grape anthocyanins. These pigments correspond to malvidin 3 glucoside linked through a vinyl bond to either (+)-catechin, (-)-epicatechin, or procyanidin dimer B3 [(+)-catechin-(+)-catechin]. NMR data of these pigments are reported for the first time. PMID- 11902965 TI - Distribution of various peptides in citrus fruits (grapefruit, lemon, and orange). AB - Tissue- and species-specific peptides of the grapefruit have been investigated by SDS-PAGE and Western blot. Five peptides from the juice and one peptide from the peel were isolated by preparative gel electrophoresis. Polyclonal antibodies were developed against them in mice. It can be established that 82, 63, and 46 kDa peptides occurred exclusively in the samples prepared from the grapefruit and the lemon juice, whereas in the orange juice, only the 82 kDa peptide could be detected. The 31 kDa peptide is characteristic for the peel samples of grapefruit and lemon. The 210 kDa peptide did not show any specificity. A 117 kDa peptide appeared in the juice and peel of grapefruit and in the peel of lemon but not in the orange. From the data of this study, it is supposed that some of the polyclonal antibodies developed against characteristic juice and peel peptides can be used to test commercial grapefruit juice products for adulteration. PMID- 11902966 TI - Development of a potentiometric method to measure the resistance to oxidation of white wines and the antioxidant power of their constituents. AB - This work describes a new potentiometric method to evaluate the resistance to oxidation of white wines. Reduction and oxidation titrations were made, and coefficient of variation obtained were 10.87 and 2.65%, respectively. The antioxidant powers of ascorbic acid (Aas) and sulfur dioxide (SO(2)) were evaluated by this method, SO(2) proving to be much less active in this respect than ascorbic acid. The two agents did not demonstrate any antioxidant synergy. A relationship between oxygen present and ascorbic acid was found by the proposed method (1 mmol of O(2) <--> 0.84 mmol of Aas). This method enables the distinction of different wines on the basis of their resistance to oxidation. PMID- 11902968 TI - Hibiscus protocatechuic acid or esculetin can inhibit oxidative LDL induced by either copper ion or nitric oxide donor. AB - Oxidation of low-density lipoprotein (LDL) could increase the incidence of atherosclerosis. Previous studies have shown that copper and sodium nitroprusside (SNP) possess the ability to oxidize LDL in a dose-dependent condition. They increase the existing negative charge in LDL and increase the electrophoretic mobility. In this study, we used protocatechuic acid (PCA) and/or esculetin (ECT) to define the antioxidative activity in oxidative LDL by relative electrophoretic mobility (REM) and thiobarbituric acid-relative substances (TBARS). The data showed that ECT and PCA possessed stronger antioxidative activity than vitamin E in oxidative LDL. A previous study showed that the level of oxidative LDL can be determined by the cholesterol degradation and fragmentation of Apo B. Our results showed that Cu(2+)-mediated oxidative LDL can induce 31% cholesterol degradation and significant fragmentation of Apo B. Both PCA and ECT exhibited remarkable ability to rescue the cholesterol degradation and Apo B fragmentation. Taken together, both PCA and ECT showed strong potency to inhibit oxidative LDL induced by copper or an NO donor. Additionally, their nontoxic characteristics elevated the possibility for their use in the daily diet; and should further prevent atherosclerosis effectively. PMID- 11902967 TI - Physicochemical and functional properties of buckwheat protein product. AB - This study was conducted to compare the physicochemical and functional properties of buckwheat protein product (BWP), soy protein isolate (SPI), and casein. BWP was prepared from buckwheat flour by the method including alkaline extraction and isoelectric precipitation. The amino acid composition of BWP was very similar to that of buckwheat flour. The protein solubility (PS) of BWP was much greater than that of SPI at all pH levels (pH 2-10) but lower than that of casein at pH 7-10. The isoelectric point of BWP was around pH 4. The higher aromatic hydrophobicities (ARH) of BWP, SPI, and casein were obtained at lower pH levels (pH 2-3). The emulsifying stability (ES) of BWP was lower than those of SPI and casein at high pH levels (pH 7-10). At all pH levels, BWP formed a thin emulsion. Regression analysis showed that the ARH of BWP was significantly associated with the ES. Although the water holding capacity of BWP was quite lower than that of SPI, its fat absorption capacity was slightly higher than those of SPI and casein. These results indicated that the physicochemical properties of BWP were different from those of SPI or casein. Thus, BWP is a potential source of functional protein for possible food application. PMID- 11902970 TI - Synthesis of theaflavin from epicatechin and epigallocatechin by plant homogenates and role of epicatechin quinone in the synthesis and degradation of theaflavin. AB - Oxidation products of (-)-epicatechin and (-)-epigallocatechin by treatment with homogenates of 62 plants belonging to 49 families were compared. Forty-six plants were capable of synthesizing theaflavin, a black tea pigment, regardless of whether they contained catechins. Loquat, Japanese pear, and blueberry had activities higher than that of fresh tea leaves after 5 h of treatment; furthermore, these plants oxidized theaflavin to theanaphthoquinone. An additional new metabolite, dehydrotheasinensin, was generated on treatment with fresh tea leaves, eggplant, and unripened Japanese orange. Evidence for the oxidation of epigallocatechin and theaflavin by electron transfer to epicatechin quinone was demonstrated in a time course study using bananas and trapping the quinone intermediates as glutathione conjugates. PMID- 11902969 TI - Repression of the 14-3-3 gene affects the amino acid and mineral composition of potato tubers. AB - Recently, transgenic potato plants were created showing underexpression of the 20R isoform of the 14-3-3 protein. The transgenic plants grown in tissue culture showed a significant increase in nitrate reductase activity and a decrease in nitrate level. The transgenic line with the lowest 14-3-3 quantity was field trialed (1997-2000) and analyzed. The reduction in the 14-3-3 protein level consistently resulted in a starch content increase and in an increase in the ratio of soluble sugars to starch in the tubers, although the latter was only barely visible. The determination of amino acid composition in the tubers showed a significant increase in methionine, proline, and arginine content and a slight but consistent increase in hydrophobic amino acid and lysine content in the cells of the transgenic potato plants. We also observed an increase in the crude protein content, from 19 to 22.1% of the control value in consecutive years. It is proposed that all of these changes might have resulted from the downregulation of nitrate reductase and sucrose phosphate synthase activities by 14-3-3, although other potential mechanisms cannot be excluded (e.g., an increase in enzyme protein level). 14-3-3-repressed transgenic plants showed a significant increase in calcium content in their tubers. It is thus proposed that a function of the isolated 14-3-3 isoform is in the control of amino acid synthesis and calcium metabolism. However, the mechanism of this control is as yet unknown. PMID- 11902971 TI - Relationship between cyanogenic compounds in kernels, leaves, and roots of sweet and bitter kernelled almonds. AB - The relationship between the levels of cyanogenic compounds (amygdalin and prunasin) in kernels, leaves, and roots of 5 sweet-, 5 slightly bitter-, and 5 bitter-kernelled almond trees was determined. Variability was observed among the genotypes for these compounds. Prunasin was found only in the vegetative part (roots and leaves) for all genotypes tested. Amygdalin was detected only in the kernels, mainly in bitter genotypes. In general, bitter-kernelled genotypes had higher levels of prunasin in their roots than nonbitter ones, but the correlation between cyanogenic compounds in the different parts of plants was not high. While prunasin seems to be present in most almond roots (with a variable concentration) only bitter-kernelled genotypes are able to transform it into amygdalin in the kernel. Breeding for prunasin-based resistance to the buprestid beetle Capnodis tenebrionis L. is discussed. PMID- 11902972 TI - Qualitative determination of specific protein glycation products by matrix assisted laser desorption/ionization mass spectrometry Peptide mapping. AB - The nonenzymatic reaction between reducing sugars and proteins, known as the Maillard reaction, has received increased recognition from nutritional science and medical research. The development of new analytical techniques for the detection of protein-bound Maillard products is therefore crucial. In this study, we applied peptide mapping by matrix-assisted laser desorption/ionization time-of flight mass spectrometry to investigate the formation of structurally specific Maillard products on glycated lysozyme (AGE-lysozyme), produced upon incubation with D-glucose. In parallel, we synthesized N(epsilon)-(carboxymethyl)lysine modified lysozyme (CML-lysozyme) and N(epsilon)-(carboxyethyl)lysine-modified lysozyme, two well-described glycation products, as model substances. 3 Deoxyglucosone-modified lysozyme and methylglyoxal-modified lysozyme were prepared as examples of glycation products incubated with dicarbonyl compounds. We were able to detect specific modifications on AGE-lysozyme, which were assigned to CML, imidazolone A, and the Amadori product. PMID- 11902974 TI - Identification of aminoethylcysteine ketimine decarboxylated dimer, a natural antioxidant, in dietary vegetables. AB - Aminoethylcysteine ketimine decarboxylated dimer (simply named dimer) is a natural sulfur-containing tricyclic compound detected, until now, in human urine, bovine cerebellum, and human plasma. Recently, the antioxidant properties of this compound have been demonstrated. In this investigation, the presence of aminoethylcysteine ketimine decarboxylated dimer was identified in garlic, spinach, tomato, asparagus, aubergine, onion, pepper, and courgette. Identification of this compound in dietary vegetables was performed using gas chromatography, high-performance liquid chromatography, and gas chromatography mass spectrometry. Results from GC analysis range in the order of 10(-4) micromol of dimer/g for all the tested vegetables. These results and the lack of a demonstrated biosynthetic pathway in humans might account for a dietary supply of this molecule. PMID- 11902973 TI - Antioxidant properties of ferulic acid and its related compounds. AB - Antioxidant activity of 24 ferulic acid related compounds together with 6 gallic acid related compounds was evaluated using several different physical systems as well as their radical scavenging activity. The radical scavenging activity on 1,1 diphenyl-2-picrylhydrazyl (DPPH) decreased in the order caffeic acid > sinapic acid > ferulic acid > ferulic acid esters > p-coumaric acid. In bulk methyl linoleate, test hydroxycinnamic acids and ferulic acid esters showed antioxidant activity in parallel with their radical scavenging activity. In an ethanol-buffer solution of linoleic acid, the activity of test compounds was not always associated with their radical scavenging activity. Ferulic acid was most effective among the tested phenolic acids. Esterification of ferulic acid resulted in increasing activity. The activity of alkyl ferulates was somewhat influenced by the chain length of alcohol moiety. When the inhibitory effects of alkyl ferulates against oxidation of liposome induced by AAPH were tested, hexyl, octyl, and 2-ethyl-1-hexyl ferulates were more active than the other alkyl ferulates. Furthermore, lauryl gallate is most effective among the tested alkyl gallates. These results indicated that not only the radical scavenging activity of antioxidants, but also their affinity with lipid substrates, might be important factors in their activity. PMID- 11902975 TI - Sodium copper chlorophyllin: in vitro digestive stability and accumulation by Caco-2 human intestinal cells. AB - Sodium copper chlorophyllin (SCC), a mixture of water-soluble chlorophyll derivatives, is used as both a food colorant and a common dietary supplement. Although the potential antimutagenic and antioxidant properties of this commercial preparation have been demonstrated, limited information is available on its digestion and absorption by humans. Stability of SCC was examined during simulated gastric and small intestinal digestion. Three preparations were subjected to in vitro digestion: SCC in water, SCC in water + 10% corn oil, and SCC in applesauce. SCC components from raw material preparations and in digested samples were analyzed by C(18) HPLC with photodiode array detection. Cu(II)chlorin e(4), the major chlorin component of SCC, was relatively stable during simulated digestion. In contrast, greater than 90% of Cu(II)chlorin e(6) was degraded to undetermined products during digestion. Recovery of Cu(II)chlorin e(6) after digestion was increased by incorporation of SCC into applesauce, suggesting a protective role of the inclusion matrix for stabilization of labile SCC components. Accumulation of SCC derivatives was investigated by using differentiated cultures of the TC7 clone of the Caco-2 human intestinal cell line. Cellular accumulation from media containing 0.5 to 60 ppm SCC was linear with intracellular content ranging between 0.2 and 29.6 microg of total SCC per mg of cellular protein. Uptake of SCC by Caco-2 cells was significantly (p < 0.01) lower in cultures incubated at 4 degrees C than in those incubated at 37 degrees C. Although intracellular SCC was transported into both apical and basolateral compartments when Caco-2 cells were grown on inserts, apical efflux was significantly greater (p < 0.01) than basolateral efflux. Stability of Cu(II)chlorin e(4) during in vitro digestion and effective uptake by Caco-2 enterocyte-like cells support the likelihood that a portion of this SCC component or its metabolites is absorbed from the human intestine. PMID- 11902976 TI - 90-day oral toxicity study of a grape seed extract (IH636) in rats. AB - To assess the safety of grape seed extract with less than 5.5% catechin monomers (IH636), 4 groups of male and female Sprague-Dawley rats were provided grape seed extract in the diet at levels of 0 (control), 0.5, 1.0, or 2.0% for a period of 90 days. All animals survived the duration of the study, and no significant changes in clinical signs, hematological parameters, organ weights, ophthalmology evaluations, or histopathological findings were observed. A significant increase in food consumption was observed in male and female rats provided the grape seed extract diets compared to that of the control rats, especially in male rats consuming 2.0% grape seed extract. This effect was not accompanied by increases in body weight gains. Grape seed extract appeared to increase the insoluble fraction of the diet. Male rats in the high-dose group exhibited decreased serum iron levels and decreased serum iron/total iron binding capacity ratio compared to those of the controls, although all values were within historical ranges for Sprague-Dawley rats. In conclusion, administration of the grape seed extract IH636 to male and female Sprague-Dawley rats in the feed at levels of 0.5, 1.0, or 2.0% for 90 days did not induce any significant toxicological effects. PMID- 11902977 TI - Antimicrobial synergistic effect of linolenic acid and monoglyceride against Bacillus cereus and Staphylococcus aureus. AB - The antimicrobial effect of linolenic acid with or without monoglyceride (glycerol laurate or glycerol myristate) against six food-borne microorganisms was determined in broth medium. Minimum inhibitory concentration of linolenic acid on Bacillus cereus and Staphylococcus aureus was 20 and 50 ppm, respectively. The growth of B. cereus treated with linolenic acid at 10 ppm with 10 ppm monoglyceride was more inhibitory than that of linolenic acid alone, and the viable cell population was reduced 2-4 log cycles compared to that of the control. When linolenic acid was added at that level, the adenosine triphosphate (ATP) concentration of extracellular fluid was drastically increased compared with that of the control, and the combined effect with monoglyceride was higher than that with linolenic acid alone. However, the intracellular ATP concentration decreased compared with that of the control. From these results, we concluded that linolenic acid has a strong antimicrobial activity against B. cereus and S. aureus, and that linolenic acid combined with monoglyceride showed stronger antimicrobial activity than using linolenic acid alone. PMID- 11902978 TI - Free radical studies of ellagic acid, a natural phenolic antioxidant. AB - Ellagic acid, a plant-derived polyphenol, inhibits gamma-radiation (hydroxyl radical) induced lipid peroxidation in rat liver microsomes in a dose- and concentration-dependent manner. Its antioxidant capacity has been estimated using the 1,1-diphenyl-2-picrylhydrazyl radical assay. To understand the actual mechanisms involved in antioxidant activity and the free radical scavenging ability,a nanosecond pulse radiolysis technique has been employed. The rate constants for the reactions of several reactive oxygen species and reactive nitrogen species such as hydroxyl, peroxyl, and nitrogen dioxide radicals have been found to be in the range of 10(6)-10(9) M(-1) s(-1). The ellagic acid radicals have been characterized by the absorption spectra and decay kinetics. Studies on the reactions of ellagic acid with the 2,2'-azinobis(3 ethylbenzothiazoline-6-sulfonate) radical and the radicals of ellagic acid with ascorbate have been used to estimate its one-electron reduction potential. Ellagic acid has also been found to be a good scavenger of peroxynitrite. Using stopped-flow reaction analyzer with absorption detection, the rate constant for this reaction has been determined to be 3.7 x 10(3) M(-1) s (-1). The electron spin resonance spectra of the oxidized ellagic acid radicals have been recorded by horseradish peroxidase and hydrogen peroxide method. PMID- 11902981 TI - Scald abuse. PMID- 11902979 TI - Treatment of an ulcerated hemangioma with recombinant platelet-derived growth factor. PMID- 11902982 TI - Apathy and meteors. PMID- 11902983 TI - Phase 1 and 2 trial of bexarotene gel for skin-directed treatment of patients with cutaneous T-cell lymphoma. AB - OBJECTIVE: To evaluate the safety, dose tolerance, and efficacy of topical bexarotene gel in patients with early-stage cutaneous T-cell lymphoma (CTCL). DESIGN: Phase 1 and 2, open-label, dose-escalation clinical trial of bexarotene gel. SETTING: Three university-based clinics. PARTICIPANTS: Sixty-seven adults with early-stage (TNM stages IA-IIA) CTCL. INTERVENTIONS: Bexarotene gel, 0.1%, 0.5%, and 1.0%, applied in incremental dose adjustments from 0.1% gel every day to 1.0% gel 4 times daily or the maximal tolerated dose. MAIN OUTCOME MEASURES: Patients were followed for efficacy and safety, and treatment continued as long as they benefited. Response (> or =50% improvement) was evaluated by the Physician's Global Assessment of cutaneous disease and by an overall severity assessment of cutaneous disease, including signs of CTCL and area involved. RESULTS: Most patients tolerated topical bexarotene at 1% gel twice daily for routine use. Adverse events were generally mild to moderate in severity and were confined to treatment sites. Treatment-limiting toxic effects were associated with skin irritation and increased with gel exposure. Patients achieved an overall response rate of 63% and a clinical complete response rate of 21%. Median projected time to onset of response was 20.1 weeks (range, 4.0-86.0 weeks), and the estimated median response duration from the start of therapy was 99 weeks. Patients with no previous therapy for mycosis fungoides responded at a higher rate (75%) than those who previously underwent topical therapies (67%). CONCLUSIONS: Bexarotene gel was well tolerated, was easily self-applied, and had a substantial response rate in treating patients with early-stage CTCL. PMID- 11902984 TI - Association of dissatisfaction with care and psychiatric morbidity with poor treatment compliance. AB - OBJECTIVES: To examine factors associated with compliance with dermatologic treatment. DESIGN: Longitudinal study. Quality of life and psychological well being were measured before the dermatologic visit with a self-completed questionnaire. Telephone interviews were performed 3 days and 4 weeks after the visit to evaluate patient satisfaction and medication adherence, respectively. SETTING: Outpatient clinics of a large dermatologic hospital in Rome, Italy. PATIENTS: A total of 1389 outpatients were contacted and 722 (52%) agreed to participate. Among them, 424 responded to the inclusion criteria and were enrolled in the study. Of these, 396 (93%) completed the telephone interviews. MAIN OUTCOME MEASURE: Self-reported compliance with dermatologic treatment. RESULTS: The dermatologists' prescriptions were not exactly followed by 44% of patients. In multiple logistic regression analysis, treatment adherence was strongly associated with complete satisfaction. Poor quality of life on the emotions scale (indicating mainly high levels of shame and embarrassment) was also associated with medication adherence. On the contrary, a strong negative association was observed between psychiatric morbidity and compliance. CONCLUSIONS: This is the first longitudinal study on dermatologic patients showing that dissatisfaction with care and psychiatric morbidity are significantly and independently associated with poor medication adherence. To improve medication adherence, particular attention should be dedicated to the physician's interpersonal skills, which emerged as a major component of patient satisfaction. Moreover, our results highlight the need for a timely identification and appropriate management of psychiatric disorders in everyday dermatologic practice. PMID- 11902985 TI - Vascular inflammation (vasculitis) in sweet syndrome: a clinicopathologic study of 28 biopsy specimens from 21 patients. AB - BACKGROUND: Sweet syndrome is characterized by painful, erythematous plaques of rapid onset accompanied by fever. Absence of vasculitis is a histologic criterion for diagnosis. However, recent reports suggest that vasculitis should not exclude the diagnosis. We hypothesized that vasculitis can occur in Sweet syndrome and that it represents an epiphenomenon rather than a primary immune-mediated process. DESIGN: Skin biopsy specimens from patients with Sweet syndrome were reviewed to determine the prevalence of vasculitis. The clinicopathologic features of cases with vasculitis were evaluated for statistically significant associations. Specimens with vasculitis underwent immunofluorescence staining. SETTING: University department of dermatology, university hospital, and private practice. PATIENTS: Medical records and biopsy specimens of 21 patients meeting diagnostic criteria for Sweet syndrome were reviewed. INTERVENTIONS: None. RESULTS: The prevalence of vasculitis was 29% (6 of 21 patients). There was a significant association of vasculitis with lesions of longer duration (P =.02). Vascular immunoglobulin and complement could not be demonstrated in cases of Sweet syndrome with vasculitis. CONCLUSIONS: Vasculitis is not a primary, immune mediated process in Sweet syndrome but occurs secondary to noxious products released from neutrophils. Blood vessels in lesions of longer duration are more likely to develop vasculitis than those of shorter duration because of prolonged exposure to noxious metabolites. Vasculitis does not exclude a diagnosis of Sweet syndrome. PMID- 11902986 TI - Long-term effectiveness of treatment with terbinafine vs itraconazole in onychomycosis: a 5-year blinded prospective follow-up study. AB - OBJECTIVE: To examine long-term cure and relapse rates after treatment with continuous terbinafine and intermittent itraconazole in onychomycosis. DESIGN: Long-term prospective follow-up study. SETTING: Three centers in Iceland. SUBJECTS: The study population comprised 151 patients aged 18 to 75 years with a clinical and mycological diagnosis of dermatophyte toenail onychomycosis. INTERVENTIONS: In a double-blind, double-dummy study, patients were randomized to receive either terbinafine (250 mg/d) for 12 or 16 weeks or itraconazole (400 mg/d) for 1 week in every 4 for 12 or 16 weeks (first intervention). Patients who did not achieve clinical cure at month 18 or experienced relapse or reinfection were offered an additional course of terbinafine (second intervention). MAIN OUTCOME MEASURES: The primary efficacy criterion was mycological cure, defined as negative results on microscopy and culture at the end of follow-up and no requirement of second intervention treatment. Secondary efficacy criteria included clinical cure without second intervention treatment and mycological and clinical relapse rates. RESULTS: Median duration of follow-up was 54 months. At the end of the study, mycological cure without second intervention treatment was found in 34 (46%) of the 74 terbinafine-treated subjects and 10 (13%) of the 77 itraconazole-treated subjects (P<.001). Mycological and clinical relapse rates were significantly higher in itraconazole vs terbinafine-treated patients (53% vs 23% and 48% vs 21%, respectively). Of the 72 patients who received subsequent terbinafine treatment, 63 (88%) achieved mycological cure and 55 (76%) achieved clinical cure. CONCLUSION: In the treatment of onychomycosis, continuous terbinafine provided superior long-term mycological and clinical efficacy and lower rates of mycological and clinical relapse compared with intermittent itraconazole. PMID- 11902987 TI - Neutrophilic dermatosis (pustular vasculitis) of the dorsal hands: a report of 7 cases and review of the literature. AB - BACKGROUND: Neutrophilic dermatosis (pustular vasculitis) of the dorsal hands is a recently described disorder, which may clinically resemble a localized variant of Sweet syndrome. OBJECTIVES: To describe the clinical and histopathologic characteristics of this rare disorder; to compare and contrast these features with those of Sweet syndrome; and to investigate possible associations with systemic diseases. OBSERVATIONS: Seven women were referred for pustular or ulcerative plaques and nodules on the dorsal hands. In most patients, the initial diagnosis was cutaneous infection, but antibiotic therapy was ineffective. Skin biopsy specimens showed dense dermal neutrophilic infiltrates with leukocytoclasis and fibrinoid vascular necrosis. Cutaneous cultures yielded negative findings in all cases. Prednisone and dapsone appeared to be helpful, but recurrences were common. Minocycline hydrochloride was of uncertain benefit. Among the 7 patients, possible systemic associations included bowel disorders and a urinary tract infection. CONCLUSIONS: Neutrophilic dermatosis of the dorsal hands may be closely related to Sweet syndrome but frequently shows the histologic pattern of leukocytoclastic vasculitis. Recognition of this disorder is important, because it may be misdiagnosed as a localized cutaneous infection. Additional studies are needed to investigate further the possible associations with internal diseases, especially bowel disorders. PMID- 11902988 TI - The first international consensus on mucous membrane pemphigoid: definition, diagnostic criteria, pathogenic factors, medical treatment, and prognostic indicators. AB - OBJECTIVE: We aimed to develop consensus-based recommendations for streamlining medical communication among various health care professionals, to improve accuracy of diagnosis and treatment, and to facilitate future investigations for mucous membrane pemphigoid. PARTICIPANTS: Because of the highly specific nature of this group of diseases, the 26 invited participants included either international scholars in the field of mucous membrane pemphigoid or experts in cutaneous pharmacology representing the 3 medical disciplines ophthalmology, oral medicine, and dermatology. EVIDENCE: The first author (L.S.C.) conducted a literature search. Based on the information obtained, international experts who had contributed to the literature in the clinical care, diagnosis, and laboratory investigation for mucous membrane pemphigoid were invited to participate in a consensus meeting aimed at developing a consensus statement. CONSENSUS PROCESS: A consensus meeting was convened and conducted on May 10, 1999, in Chicago, Ill, to discuss the relevant issues. The first author drafted the statement based on the consensus developed at the meeting and the participants' written comments. The draft was submitted to all participants for 3 separate rounds of review, and disagreements were reconciled based on literature evidence. The third and final statement incorporated all relevant evidence obtained in the literature search and the consensus developed by the participants. The final statement was approved and endorsed by all 26 participants. CONCLUSIONS: Specific consensus-based recommendations were made regarding the definition, diagnostic criteria, pathogenic factors, medical treatment, and prognostic indicators for mucous membrane pemphigoid. A system of standard reporting for these patients was proposed to facilitate a uniform data collection. PMID- 11902989 TI - Interventions for mucous membrane pemphigoid/cicatricial pemphigoid and epidermolysis bullosa acquisita: a systematic literature review. AB - OBJECTIVE: To identify and critically evaluate evidence from randomized controlled trials for the efficacy of treatments for mucous membrane pemphigoid (MMP)/cicatricial pemphigoid (CP) and epidermolysis bullosa acquisita (EBA). SEARCH STRATEGY: Review of MEDLINE from 1966 through March 2000, EMBASE from 1980 through March 2000, and the Cochrane Controlled Trials Register (February 28, 2001) to identify randomized controlled trials for the efficacy of treatments in MMP/CP and EBA. SELECTION CRITERIA: All randomized controlled trials of therapeutic interventions that included patients with MMP/CP or EBA confirmed by immunofluorescence study findings. All age groups were included. RESULTS: We found 2 small randomized controlled trials of MMP/CP, both conducted in patients with severe eye involvement. We were not able to identify a randomized controlled trial of therapeutic interventions in EBA. CONCLUSIONS: There is evidence from 2 small trials that severe ocular CP responds best to treatment with cyclophosphamide, and mild to moderate disease seems effectively suppressed by treatment with dapsone. No treatment recommendations can be made for EBA because to our knowledge no randomized controlled trials are published. Even though systemic corticosteroids are regarded as the gold standard in the treatment of MMP/CP and EBA, there is poor evidence from the literature that they are the best treatment for these diseases. PMID- 11902990 TI - A systematic review of treatments for bullous pemphigoid. AB - OBJECTIVE: To assess the effectiveness of treatments for bullous pemphigoid. METHODS: The Cochrane Library search strategy was used to identify randomized controlled trials from MEDLINE and EMBASE, from their inception to September 30, 2001. All randomized controlled trials on interventions for bullous pemphigoid, confirmed by immunofluorescence studies, were included. RESULTS: We found 6 randomized controlled trials with a total of 293 patients. Two trials, one comparing prednisolone, 0.75 mg/kg per day, with prednisolone, 1.25 mg/kg per day, and the other comparing methylprednisolone with prednisolone, did not find any significant difference in effectiveness. The higher dose of prednisolone, however, was associated with more severe adverse effects. Combination treatments of prednisone with azathioprine in one trial and of prednisolone with plasma exchange in another were useful in halving the corticosteroid dose required (mean +/- SD, 0.52 +/- 0.28 mg/kg in the plasma exchange-treated group vs 0.97 +/- 0.33 mg/kg in the prednisolone only-treated group). However, a fifth trial, including all 3 treatment groups (prednisolone alone, prednisolone and azathioprine, and prednisolone and plasma exchange), failed to confirm the benefit of combination treatment over prednisolone alone. A trial of 20 patients, comparing prednisone with tetracycline and niacinamide, found no statistically significant difference in response between the 2 groups, but the prednisone-treated group had more serious adverse effects. CONCLUSIONS: There is inadequate evidence for a recommendation of a specific treatment for bullous pemphigoid, and there is a need for larger randomized controlled trials with adequate power. Starting doses of prednisolone greater than 0.75 mg/kg per day do not seem to give additional benefit, and it seems that lower doses may be adequate for disease control. The effectiveness of the addition of plasma exchange or azathioprine to corticosteroids has not been established. Combination treatment with tetracycline and niacinamide seems useful, although this needs further validation. PMID- 11902991 TI - Prevention of atopic eczema: a dream not so far away? PMID- 11902992 TI - Doctor's orders: rethinking compliance in dermatology. PMID- 11902993 TI - Bexarotene gel: a Food and Drug Administration-approved skin-directed therapy for early-stage cutaneous T-cell lymphoma. PMID- 11902995 TI - Treatment considerations while awaiting the ideal bullous pemphigoid trial. PMID- 11902996 TI - Horny excrescences on the hands. PMID- 11902994 TI - Skin lesions of Sweet syndrome and its dorsal hand variant contain vasculitis: an oxymoron or an epiphenomenon? PMID- 11902997 TI - Recurrent annular erythematous scaly patches. PMID- 11902998 TI - Chalky-yellow nodules on a neonate. PMID- 11902999 TI - Skin injury after electroencephalography. PMID- 11903000 TI - A comment on "Butcher's warts: dermatological heritage or testable misinformation?". PMID- 11903001 TI - In search of living frozen adipocytes? PMID- 11903003 TI - Cutaneous small vessel vasculitis: an entity with frequent renal involvement. PMID- 11903004 TI - Nail salons can be risky business. PMID- 11903010 TI - Surprise finding spurs Ebola researchers' hopes. PMID- 11903011 TI - Epidemic of obesity expands its spread to developing countries. PMID- 11903012 TI - Sweeping new congressional plan to fight cancer is previewed. PMID- 11903019 TI - Does high intake of vitamin A pose a risk for osteoporotic fracture? PMID- 11903017 TI - First-line vs second-line antibiotics for treatment of sinusitis. PMID- 11903021 TI - Meningiomas in women with lymphangioleiomyomatosis. PMID- 11903023 TI - Partnerships between universities and industry. PMID- 11903024 TI - Partnerships between universities and industry. PMID- 11903026 TI - Nosocomial transmission of vancomycin-resistant enterococci from surfaces. PMID- 11903027 TI - Cognitive outcome after off-pump and on-pump coronary artery bypass graft surgery: a randomized trial. AB - CONTEXT: Coronary artery bypass graft (CABG) surgery is associated with a decline in cognitive function, which has largely been attributed to the use of cardiopulmonary bypass (on-pump procedures). Cardiac stabilizers facilitate CABG surgery without use of cardiopulmonary bypass (off-pump procedures) and should reduce the cognitive decline associated with on-pump procedures. OBJECTIVE: To compare the effect of CABG surgery with (on-pump) and without (off-pump) cardiopulmonary bypass on cognitive outcome. DESIGN AND SETTING: Randomized controlled trial conducted in the Netherlands of CABG surgery patients enrolled from March 1998 through August 2000, with 3- and 12-month follow-up. PARTICIPANTS AND INTERVENTION: Patients scheduled for their first CABG surgery (mean age, 61 years; n = 281) were randomly assigned to off-pump surgery (n = 142) or on-pump surgery (n = 139). MAIN OUTCOME MEASURES: Cognitive outcome at 3 and 12 months, which was determined by psychologists (blinded for randomization) who administered 10 neuropsychological tests before and after surgery. Quality of life, stroke rate, and all-cause mortality at 3 and 12 months were secondary outcome measures. RESULTS: Cognitive outcome could be determined at 3 months in 248 patients. Cognitive decline occurred in 21% in the off-pump group and 29% in the on-pump group (relative risk [RR], 0.65; 95% confidence interval [CI], 0.36 1.16; P =.15). The overall standardized change score (ie, improvement of cognitive performance) was 0.19 in the off-pump vs 0.13 in the on-pump group (P =.03). At 12 months, cognitive decline occurred in 30.8% in the off-pump group and 33.6% in the on-pump group (RR, 0.88; 95% CI, 0.52-1.49; P =.69). The overall standardized change score was 0.19 in the off-pump vs 0.12 in the on-pump group (P =.09). No statistically significant differences were observed between the on pump and off-pump groups in quality of life, stroke rate, or all-cause mortality at 3 and 12 months. CONCLUSION: Patients who received their first CABG surgery without cardiopulmonary bypass had improved cognitive outcomes 3 months after the procedure, but the effects were limited and became negligible at 12 months. PMID- 11903028 TI - Risk of non-Hodgkin lymphoma in celiac disease. AB - CONTEXT: Celiac disease is one of the most common lifelong disorders. Non-Hodgkin lymphoma is a possible complication of celiac disease and may lead to a large portion of lymphoma cases. OBJECTIVE: To quantify the risk for developing non Hodgkin lymphoma of any primary site associated with celiac disease. DESIGN AND SETTING: Multicenter, case-control study conducted between January 1996 and December 1999 throughout Italy. PATIENTS: Cases were older than 20 years (median, 57; range, 20-92 years) with non-Hodgkin lymphoma of any primary site and histological type and were recruited at the time of the diagnosis. Controls were healthy adults (2739 men and 2981 women) from the general population. MAIN OUTCOME MEASURE: Positive test result for class A serum antiendomysial antibody. RESULTS: Celiac disease was diagnosed in 6 (0.92%) of 653 patients with lymphoma. Of the 6 cases, 3 were of B-cell and 3 were of T-cell origin. Four of 6 cases had lymphoma primarily located in the gut. In the control group, 24 (0.42%) had celiac disease. The odds ratio (adjusted for age and sex) for non-Hodgkin lymphoma of any primary site associated with celiac disease was 3.1 (95% confidence interval [CI], 1.3-7.6), 16.9 (95% CI, 7.4-38.7) for gut lymphoma, and 19.2 (95% CI, 7.9-46.6) for T-cell lymphoma, respectively. The risk for non Hodgkin lymphoma for the overall population, which was adjusted for age and sex, was 0.63% (95% CI, - 0.12% to 1.37%). CONCLUSION: Celiac disease is associated with an increased risk for non-Hodgkin lymphoma, especially of T-cell type and primarily localized in the gut. However, the association does not represent a great enough risk to justify early mass screening for celiac disease. PMID- 11903029 TI - Relationship between insulin resistance and an endogenous nitric oxide synthase inhibitor. AB - CONTEXT: Increased levels of asymmetric dimethylarginine (ADMA) are associated with endothelial dysfunction and increased risk of cardiovascular disease. Several cardiovascular risk factors are associated with reduced sensitivity to insulin, but elevated ADMA concentrations have not been fully linked to the metabolic syndrome. OBJECTIVE: To evaluate the relationship between insulin sensitivity and plasma ADMA concentrations, and to determine whether a pharmacological treatment that increases insulin sensitivity would also modulate ADMA concentrations. DESIGN, SETTING, AND SUBJECTS: Cross-sectional study, containing a nonrandomized controlled trial component, of 64 healthy volunteers without diabetes (42 women, 22 men; 48 with normal blood pressure and 16 with hypertension), which was conducted at a university medical center between October 2000 and July 2001. INTERVENTION: Rosiglitazone (4 mg/d for 4 weeks and then 4 mg twice daily for 8 weeks), an insulin-sensitizing agent, was given to 7 insulin resistant subjects with hypertension. These subjects were studied before and after 12-week treatment. MAIN OUTCOME MEASURES: Insulin sensitivity measured by the insulin suppression test, and fasting plasma levels of low-density lipoprotein cholesterol, triglycerides, high-density lipoprotein cholesterol, glucose, insulin, and ADMA concentrations. RESULTS: Plasma ADMA concentrations were positively correlated with impairment of insulin-mediated glucose disposal in nondiabetic, normotensive subjects (r = 0.73; P<.001). Consistent with the metabolic syndrome, ADMA levels were also positively correlated with fasting triglyceride levels (r = 0.52; P<.001) but not with low-density lipoprotein cholesterol levels (r = 0.19; P =.20). Plasma ADMA concentrations increased in insulin-resistant subjects independent of hypertension. Pharmacological treatment improved insulin sensitivity and reduced mean (SD) plasma ADMA concentrations from 1.50 (0.30) to 1.05 (0.33) micromol/L (P =.001). CONCLUSION: A significant relationship exists between insulin resistance and plasma concentrations of ADMA. Pharmacological intervention with rosiglitazone enhanced insulin sensitivity and reduced ADMA levels. Increases in plasma ADMA concentrations may contribute to the endothelial dysfunction observed in insulin-resistant patients. PMID- 11903030 TI - Biochemical diagnosis of pheochromocytoma: which test is best? AB - CONTEXT: Diagnosis of pheochromocytoma depends on biochemical evidence of catecholamine production by the tumor. However, the best test to establish the diagnosis has not been determined. OBJECTIVE: To determine the biochemical test or combination of tests that provides the best method for diagnosis of pheochromocytoma. DESIGN, SETTING, AND PARTICIPANTS: Multicenter cohort study of patients tested for pheochromocytoma at 4 referral centers between 1994 and 2001. The analysis included 214 patients in whom the diagnosis of pheochromocytoma was confirmed and 644 patients who were determined to not have the tumor. MAIN OUTCOME MEASURES: Test sensitivity and specificity, receiver operating characteristic curves, and positive and negative predictive values at different pretest prevalences using plasma free metanephrines, plasma catecholamines, urinary catecholamines, urinary total and fractionated metanephrines, and urinary vanillylmandelic acid. RESULTS: Sensitivities of plasma free metanephrines (99% [95% confidence interval [CI], 96%-100%]) and urinary fractionated metanephrines (97% [95% CI, 92%-99%]) were higher than those for plasma catecholamines (84% [95% CI, 78%-89%]), urinary catecholamines (86% [95% CI, 80%-91%]), urinary total metanephrines (77% [95% CI, 68%-85%]), and urinary vanillylmandelic acid (64% [95% CI, 55%-71%]). Specificity was highest for urinary vanillylmandelic acid (95% [95% CI, 93%-97%]) and urinary total metanephrines (93% [95% CI, 89%-97%]); intermediate for plasma free metanephrines (89% [95% CI, 87%-92%]), urinary catecholamines (88% [95% CI, 85%-91%]), and plasma catecholamines (81% [95% CI, 78%-84%]); and lowest for urinary fractionated metanephrines (69% [95% CI, 64% 72%]). Sensitivity and specificity values at different upper reference limits were highest for plasma free metanephrines using receiver operating characteristic curves. Combining different tests did not improve the diagnostic yield beyond that of a single test of plasma free metanephrines. CONCLUSION: Plasma free metanephrines provide the best test for excluding or confirming pheochromocytoma and should be the test of first choice for diagnosis of the tumor. PMID- 11903031 TI - beta-Blockers and reduction of cardiac events in noncardiac surgery: scientific review. AB - CONTEXT: Recent studies suggest that perioperatively administered beta-blockers may reduce the risk of adverse cardiac events in patients undergoing major noncardiac surgery. OBJECTIVE: To review the efficacy of perioperative beta blockade in reducing myocardial ischemia, myocardial infarction, and cardiac or all-cause mortality from randomized trials. DATA SOURCES: A MEDLINE and conventional search of English-language articles published since 1980 was performed to gather information related to perioperative cardiac complications and beta-blockade. Reference lists from all relevant articles and published recommendations for perioperative cardiac risk management were reviewed to identify additional studies. STUDY SELECTION AND DATA EXTRACTION: Prospective randomized studies (6) were included in the analysis if they discussed the impact of beta-blockade on perioperative cardiac ischemia, myocardial infarction, and mortality for patients undergoing major noncardiac surgery. Articles were examined for elements of trial design, treatment protocols, important biases, and major findings. These elements were then qualitatively compared. DATA SYNTHESIS: We identified 5 randomized controlled trials: 4 assessed myocardial ischemia and 3 reported myocardial infarction, cardiac, or all-cause mortality. All studies sought to achieve beta-blockade before the induction of anesthesia by titrating doses to a target heart rate. Of studies reporting myocardial ischemia, numbers needed to treat were modest (2.5-6.7). Similarly modest numbers needed to treat were observed in studies reporting a significant impact on cardiac or all-cause mortality (3.2-8.3). The most marked effects were seen in patients at high risk; the sole study reporting a nonsignificant result enrolled patients with low baseline risk. As a group, studies of perioperative beta-blockade have enrolled relatively few carefully selected patients. In addition, differences in treatment protocols leave questions unanswered regarding optimal duration of therapy. CONCLUSIONS: Despite heterogeneity of trials, a growing literature suggests a benefit of beta-blockade in preventing perioperative cardiac morbidity. Evidence from these trials can be used to formulate an effective clinical approach while definitive trials are awaited. PMID- 11903032 TI - beta-Blockers and reduction of cardiac events in noncardiac surgery: clinical applications. AB - Recent studies suggest that beta-blockers administered perioperatively may reduce the risk of adverse cardiac events and mortality in patients who have cardiac risk factors and undergo major noncardiac surgery. The objective of this article is to provide practicing physicians with examples of perioperative beta-blocker use in practice by using several hypothetical cases. Although current evidence describing the effectiveness of perioperative beta-blockade may not address all possible clinical situations, it is possible to formulate an evidence-based approach that will maximize benefit to patients. We describe how information from several sources can be used to guide management of patients with limited exercise tolerance, those at highest risk for perioperative cardiac events, patients who are taking beta-blockers long-term, and those with relative contraindications to beta-blockade. Even though fine points of their use remain to be elucidated, perioperative beta-blocker use is important and can be easily applied in practice by any physician involved with the care of patients perioperatively. PMID- 11903033 TI - Protecting the brain in coronary artery bypass graft surgery. PMID- 11903034 TI - Insulin resistance, ADMA levels, and cardiovascular disease. PMID- 11903035 TI - Rotavirus vaccine and the news media, 1987-2001. AB - CONTEXT: In August 1998, the US Food and Drug Administration licensed the first vaccine against rotavirus, the most important cause of severe childhood diarrhea. Fourteen months later, amid intense media activity, the vaccine was withdrawn after an association was found with intussusception. OBJECTIVES: To examine the character of news media stories about rotavirus vaccine before and after intussusception became an issue, to evaluate what prompted the stories, and to assess the extent to which they evoked public reaction. DESIGN AND SETTING: We searched Lexis-Nexis and Video Monitoring Services of America databases for rotavirus vaccine stories from the first US clinical trials (January 1, 1987) until 17 months after withdrawal (March 31, 2001) and examined calls to the National Immunization Hotline during the period in which rotavirus vaccine information was captured (July 1-December 31, 1999). MAIN OUTCOME MEASURES: Mention of vaccine benefits and adverse events, classification of stories as positive, negative, or neutral toward the vaccine, story stimuli, and public response. RESULTS: We included 280 newspaper (primary subject of analysis), 49 wire service, and 257 television stories. Prior to identification of the intussusception association (January 1, 1987-July 14, 1999), 21% of 188 newspaper stories mentioned vaccine adverse events and only 2 stories were negative toward the vaccine. Ninety-nine percent of stories mentioned vaccine benefits. During the period surrounding withdrawal (July 15-December 31, 1999), 93% of 90 stories mentioned adverse events and 77% were negative toward the vaccine. Eighty-four percent mentioned vaccine benefits. The rate of stories per month was 14-fold greater than the preceding period (P<.001); temporal and geographic patterns of media and hotline activity were similar. Thereafter (January 1, 2000-March 31, 2001), only 2 stories focused on rotavirus vaccine. Scientific research or public health actions prompted 80% of stories. Wire service and television stories showed similar patterns. The increase in rotavirus stories in July 1999 was followed by an increase in calls to the National Immunization Hotline regarding rotavirus but not other topics. The number of rotavirus calls that month was 57% higher than for any other childhood vaccine for any month since the hotline began in 1997. Rotavirus calls ceased almost completely after withdrawal of the vaccine in October 1999. CONCLUSIONS: In response to reports about an adverse event, news media stories about vaccines can change abruptly from positivity to negativity. Since most vaccine stories may be stimulated by research and public health actions, opportunities exist to provide the media with accurate information necessary to avoid the "early idealization-sudden condemnation" pattern seen with rotavirus vaccine. PMID- 11903040 TI - Isoform-specific knockdown and expression of adaptor protein ShcA using small interfering RNA. AB - Many eukaryotic genes are expressed as multiple isoforms through the differential utilization of transcription/translation initiation sites or alternative splicing. The conventional approach for studying individual isoforms in a clean background (i.e. without the influence of other isoforms) has been to express them in cells or whole organisms in which the target gene has been deleted; this is time-consuming. Recently an efficient post-transcriptional gene-silencing method has been reported that employs a small interfering double-stranded RNA (siRNA). On the basis of this method we report a rapid alternative approach for isoform-specific gene expression. We show how the adaptor protein ShcA can be suppressed and expressed in an isoform-specific manner in a human cell line. ShcA exists in three isoforms, namely p66, p52 and p46, which differ only in their N terminal regions and are derived from two different transcripts, namely p66 and p52/p46 mRNAs. An siRNA with a sequence shared by the two transcripts suppressed all of them. However, another siRNA whose sequence was present only in p66 mRNA suppressed only the p66 isoform, suggesting that the siRNA signal did not propagate to other regions of the target mRNA. The expression of individual isoforms was achieved by first down-regulating all isoforms by the common siRNA and then transfecting with an expression vector for each isoform that harboured silent mutations at the site corresponding to the siRNA. This allowed functional analysis of individual ShcA isoforms and may be more generally applicable for studying genes encoding multiple proteins. PMID- 11903041 TI - Structure determination of the exopolysaccharide produced by Lactobacillus rhamnosus strains RW-9595M and R. AB - Exopolysaccharides (EPSs) were isolated and purified from Lactobacillus rhamnosus strains RW-9595M, which has been shown to possess cytokine-stimulating activity, and R grown under various fermentation conditions (carbon source, incubation temperature and duration). Identical (1)H NMR spectra were obtained in all cases. Molecular masses were determined by gel permeation chromatography. The primary structure was elucidated using chemical and spectroscopic techniques. Organic acid, monosaccharide and absolute configuration analyses gave the following composition: pyruvate, 1; D-glucose, 2; D-galactose, 1; and l-rhamnose, 4. Methylation analysis indicated the presence of three residues of 3-linked rhamnose, and one residue each of 2,3-linked rhamnose, 2-linked glucose, 3-linked glucose and 4,6-linked galactose. The EPS was submitted to periodate oxidation followed by borohydride reduction. Monosaccharide analysis of the resulting polysaccharide gave the new composition: rhamnose, 4; and glucose, 1. Methylation analysis confirmed the loss of the 2-linked glucose and 4,6-linked galactose residues. On the basis of one- and two-dimensional (1)H and (13)C NMR data, the structure of the native EPS was consistent with the following heptasaccharide repeating unit: [3Rha alpha-3Glc beta-3[Gal4,6(R)Py alpha-2]Rha alpha-3Rha alpha 3Rha alpha-2Glc alpha-](n) where Rha corresponds to rhamnose (6-deoxymannose) and Py corresponds to pyruvate acetal. Complete (1)H and (13)C assignments are reported for the native and the corresponding pyruvate-hydrolysed polysaccharide. Electrospray MS and MS/MS data are given for the oligosaccharide produced by Smith degradation. PMID- 11903044 TI - Human insulin-like growth factor II leader 2 mediates internal initiation of translation. AB - Insulin-like growth factor II (IGF-II) is a fetal growth factor, which belongs to the family of insulin-like peptides. During fetal life, the IGF-II gene generates three mRNAs with different 5' untranslated regions (UTRs), but identical coding regions and 3' UTRs. We have shown previously that IGF-II leader 3 mRNA translation is regulated by a rapamycin-sensitive pathway, whereas leader 4 mRNA is constitutively translated, but so far the significance of leader 2 mRNA has been unclear. Here, we show that leader 2 mRNA is translated efficiently in an eIF4E-independent manner. In a bicistronic vector system, the 411 nt leader 2 was capable of internal initiation via a phylogenetically conserved internal ribosome entry site (IRES), located in the 3' half of the leader. The IRES is composed of an approx. 120 nt ribosome recruitment element, followed by an 80 nt spacer region, which is scanned by the ribosomal pre-initiation complex. Since cap dependent translation is down-regulated during cell division, leader 2 might facilitate a continuous IGF-II production in rapidly dividing cells during development. PMID- 11903043 TI - Cyclophilin-A is involved in excitotoxin-induced caspase activation in rat neuronal B50 cells. AB - Glutamate and the NO donor, nitroprusside, synergistically induced the death of B50 cells from a rat CNS-derived neuroblastoma cell line. With low [nitroprusside] (10 microM) both nitroprusside and glutamate were required. Under these conditions, nuclei became pyknotic and caspases were activated. The activities of caspase-3 and caspase-6 (effector caspases) were higher than those of caspase-8 and caspase-9 (initiator caspases). The activation of all four caspases was inhibited by cyclosporin A, with the order of susceptibility caspase 8=caspase-9=caspase-6>caspase-3. To identify the possible locus of cyclosporin A action, we used an antisense oligodeoxynucleotide to suppress the level of cyclophilin-A to<5% of its control value. Cyclophilin-A suppression largely reproduced the inhibitory effects of cyclosporin A. These results provide the first indication that cyclophilin-A participates in the activation of the caspase cascade in neuronal cells, in particular in the form of cascade elicited by excitotoxic stimuli. It is concluded that neuroprotection by cyclosporin A against excitotoxin-induced apoptosis is, at least partly, due to inhibition of cyclophilin-A. PMID- 11903042 TI - Platelet-derived growth factor-BB-mediated glycosaminoglycan synthesis is transduced through Akt. AB - Previously we have demonstrated that the phosphoinositide 3-kinase (PI-3K) signal transduction pathway mediates platelet-derived growth factor (PDGF)-BB-induced glycosaminoglycan (GAG) synthesis in fetal lung fibroblasts. In the present study we further investigated the signal-transduction pathway(s) that results in PDGF BB-induced GAG synthesis. Over-expression of a soluble PDGF beta-receptor as well as a mutated form of the beta-receptor, unable to bind PI-3K, diminished GAG synthesis in fetal lung fibroblasts subsequent to PDGF-BB stimulation. The PI-3K inhibitor wortmannin blocked PDGF-BB-induced Akt activity as well as significantly diminishing PDGF-BB-mediated GAG synthesis. Expression of dominant negative PI-3K also abrogated Akt activity and GAG synthesis. Furthermore, expression of dominant-negative Akt abrogated endogenous Akt activity, Rab3D phosphorylation and GAG synthesis, whereas expression of constitutively activated Akt stimulated Rab3D phosphorylation and GAG synthesis in the absence of PDGF-BB. Over-expression of wild-type PTEN (phosphatase and tensin homologue deleted in chromosome 10) inhibited Akt activity and concomitantly attenuated GAG synthesis in fibroblasts stimulated with PDGF-BB. These data suggest that Akt is an integral protein involved in PDGF-BB-mediated GAG regulation in fetal lung fibroblasts. PMID- 11903045 TI - Ceramides increase the activity of the secretory phospholipase A2 and alter its fatty acid specificity. AB - Modulation of human recombinant secretory type II phospholipase A(2) activity by ceramide and cholesterol was investigated using model glycerophospholipid substrates composed of phosphatidylethanolamine and phosphatidylserine dispersed in aqueous medium. Enzyme activity was monitored by measurement of released fatty acids using capillary GC-MS. Fatty acids from the sn-2 position of the phospholipids were hydrolysed by the enzyme in proportion to the relative abundance of the phospholipid in the substrate. Addition of increasing amounts of ceramide to the substrate progressively enhanced phospholipase activity. The increased activity was accomplished largely by preferential hydrolysis of polyunsaturated fatty acids, particularly arachidonic acid, derived from phosphatidylethanolamine. The addition of sphingomyelin to the substrate glycerophospholipids inhibited phospholipase activity but its progressive substitution by ceramide, so as to mimic sphingomyelinase activity, counteracted the inhibition. The presence of cholesterol in dispersions of glycerophospholipid substrate-containing ceramides suppressed activation of the enzyme resulting from the presence of ceramide. The molecular basis of enzyme modulation was investigated by analysis of the phase structure of the dispersed lipid substrate during temperature scans from 46 to 20 degrees C using small-angle synchrotron X ray diffraction. These studies indicated that intermediate structures created after ceramide-dependent phase separation of hexagonal and lamellar phases represent the most susceptible form of the substrate for enzyme hydrolysis. PMID- 11903046 TI - An alternative model of H ferritin promoter transactivation by c-Jun. AB - c-Jun is a member of the activator protein 1 family, and its interaction with different nuclear factors generates a wide spectrum of complexes that regulate transcription of different promoters. H ferritin promoter transcription is tightly dependent on nuclear factor Y (NFY). Ferritin transcription is activated by c-Jun, although the promoter does not contain a canonical binding site. NFY, on the other hand, does not bind c-Jun in vitro, whereas in vivo c-Jun is found in the complex containing NFY. Moreover, a c-Jun-GCN4 chimaeric construct containing only the transactivation domain of Jun and the basic-region leucine zipper domain of GCN4 stimulates the H ferritin promoter. A synthetic GAL4 promoter and the cognate activator, the fusion protein NFY-GAL4, are potently activated by c-Jun. Titration of p300 by co-expressing E1A abolishes the stimulatory effect. Moreover, another p300-dependent promoter, the cAMP-response element, can be superactivated by c-Jun using the same mechanism. These data indicate that c-Jun, when activated or overexpressed, is recruited to the H ferritin promoter by p300, which links NFY, bound to DNA, to the complex. These results add a new level of complexity to transcriptional regulation by c-Jun, which can activate p300-dependent promoters without binding directly to the target DNA. PMID- 11903047 TI - Evidence that zymogen granules do not function as an intracellular Ca2+ store for the generation of the Ca2+ signal in rat parotid acinar cells. AB - Rat parotid acinar cells lacking zymogen granules were obtained by inducing granule discharge with the beta-adrenoceptor agonist isoproterenol. To assess whether zymogen granules are involved in the regulation of Ca(2+) signalling as intracellular Ca(2+) stores, changes in cytosolic free Ca(2+) ion concentration ([Ca(2+)](i)) were studied with imaging microscopy in fura-2-loaded parotid acinar cells lacking zymogen granules. The increase in [Ca(2+)](i) induced by muscarinic receptor stimulation was initiated at the apical pole of the acinar cells, and rapidly spread as a Ca(2+) wave towards the basolateral region. The magnitude of the [Ca(2+)](i) response and the speed of the Ca(2+) wave were essentially similar to those in control acinar cells containing zymogen granules. Western blot analysis of the inositol 1,4,5-trisphosphate receptor (IP(3)R) was performed on zymogen granule membranes and microsomes using anti-IP(3)R antibodies. The immunoreactivity of all three IP(3)Rs was clearly observed in the microsomal preparations. Although a weak band of IP(3)R type-2 was detected in the zymogen granule membranes, this band probably resulted from contamination by the endoplasmic reticulum (ER), because calnexin, a marker protein of the ER, was also detected in the same preparation. Furthermore, Western blotting and reverse transcriptase-PCR analysis failed to provide evidence for the expression of ryanodine receptors in rat parotid acinar cells, whereas expression was clearly detectable in rat skeletal muscle, heart and brain. These results suggest that zymogen granules do not have a critical role in Ca(2+) signalling in rat parotid acinar cells. PMID- 11903048 TI - Investigation of the role of Endo180/urokinase-type plasminogen activator receptor-associated protein as a collagenase 3 (matrix metalloproteinase 13) receptor. AB - Procollagenase 3 can be activated by interaction with and cleavage by the cell associated membrane type 1 metalloproteinase (MT1 MMP; MMP 14). It has also been shown to bind to a specific receptor, and is subsequently internalized via the low-density lipoprotein-related receptor by osteoblast cell lines. The receptor was identified as a recycling glycoprotein of the macrophage mannose receptor family, Endo180. In order to ascertain whether there is a relationship between Endo180 binding and procollagenase 3 activation, we have compared procollagenase 3 activation by an HT1080 fibrosarcoma cell line overexpressing MT1 MMP, without and with overexpression of Endo180. No difference in procollagenase 3 activation was observed, and neither was the enzyme bound to the cells or internalized. In contrast, the osteoblast cell lines, MG63 and UMR-106, both bound and internalized procollagenase 3. However, immunolocalization studies showed that the Endo180 abundantly expressed by these cells did not co-localize with the procollagenase 3. In further biochemical studies we confirmed that procollagenase 3 did not bind to Endo180, using both ligand- blotting and immunoprecipitation techniques. We conclude that Endo180 is unlikely to be a receptor for collagenase 3 in relation to either its activation or cell binding and internalization, and that other interaction partners must be sought. PMID- 11903050 TI - Expression, purification and characterization of human glutamate dehydrogenase (GDH) allosteric regulatory mutations. AB - Glutamate dehydrogenase (GDH) catalyses the reversible oxidative deamination of l glutamate to 2-oxoglutarate in the mitochondrial matrix. In mammals, this enzyme is highly regulated by allosteric effectors. The major allosteric activator and inhibitor are ADP and GTP, respectively; allosteric activation by leucine may play an important role in amino acid-stimulated insulin secretion. The physiological significance of this regulation has been highlighted by the identification of children with an unusual hyperinsulinism/hyperammonaemia syndrome associated with dominant mutations in GDH that cause a loss in GTP inhibition. In order to determine the effects of these mutations on the function of the human GDH homohexamer, we studied the expression, purification and characterization of two of these regulatory mutations (H454Y, which affects the putative GTP-binding site, and S448P, which affects the antenna region) and a mutation designed to alter the putative binding site for ADP (R463A). The sensitivity to GTP inhibition was impaired markedly in the purified H454Y (ED(50), 210 microM) and S448P (ED(50), 3.1 microM) human GDH mutants compared with the wild-type human GDH (ED(50), 42 nM) or GDH isolated from heterozygous patient cells (ED(50), 290 and 280 nM, respectively). Sensitivity to ADP or leucine stimulation was unaffected by these mutations, confirming that they interfere specifically with the inhibitory GTP-binding site. Conversely, the R463A mutation completely eliminated ADP activation of human GDH, but had little effect on either GTP inhibition or leucine activation. The effects of these three mutations on ATP regulation indicated that this nucleotide inhibits human GDH through binding of its triphosphate tail to the GTP site and, at higher concentrations, activates the enzyme through binding of the nucleotide to the ADP site. These data confirm the assignment of the GTP and ADP allosteric regulatory sites on GDH based on X-ray crystallography and provide insight into the structural mechanisms involved in positive and negative allosteric control and in inter-subunit co-operativity of human GDH. PMID- 11903049 TI - Monitoring of exocytosis and endocytosis of insulin secretory granules in the pancreatic beta-cell line MIN6 using pH-sensitive green fluorescent protein (pHluorin) and confocal laser microscopy. AB - The dynamics of exocytosis/endocytosis of insulin secretory granules in pancreatic beta-cells remains to be clarified. In the present study, we visualized and analysed the motion of insulin secretory granules in MIN6 cells using pH-sensitive green fluorescent protein (pHluorin) fused to either insulin or the vesicle membrane protein, phogrin. In order to monitor insulin exocytosis, pHluorin, which is brightly fluorescent at approximately pH 7.4, but not at approximately pH 5.0, was attached to the C-terminus of insulin. To monitor the motion of insulin secretory granules throughout exocytosis/endocytosis, pHluorin was inserted between the third and fourth amino acids after the identified signal peptide cleavage site of rat phogrin cDNA. Using this method of cDNA construction, pHluorin was located in the vesicle lumen, which may enable discrimination of the unfused acidic secretory granules from the fused neutralized ones. In MIN6 cells expressing insulin-pHluorin, time-lapse confocal laser scanning microscopy (5 or 10 s intervals) revealed the appearance of fluorescent spots by depolarization after stimulation with 50 mM KCl and 22 mM glucose. The number of these spots in the image at the indicated times was counted and found to be consistent with the results of insulin release measured by RIA during the time course. In MIN6 cells expressing phogrin-pHluorin, data showed that fluorescent spots appeared following high KCl stimulation and remained stationary for a while, moved on the plasma membrane and then disappeared. Thus we demonstrate the visualized motion of insulin granule exocytosis/endocytosis using the pH-sensitive marker, pHluorin. PMID- 11903051 TI - Solute carrier 11a1 (Slc11a1; formerly Nramp1) regulates metabolism and release of iron acquired by phagocytic, but not transferrin-receptor-mediated, iron uptake. AB - Solute carrier 11a1 (Slc11a1; formerly Nramp1; where Nramp stands for natural resistance-associated macrophage protein) is a proton/bivalent cation antiporter that localizes to late endosomes/lysosomes and controls resistance to pathogens. In the present study the role of Slc11a1 in iron turnover is examined in macrophages transfected with Slc11a1(Gly169) (wild-type) or Slc11a1(Asp169) (mutant=functional null) alleles. Following direct acquisition of transferrin (Tf)-bound iron via the Tf receptor, iron uptake and release was equivalent in wild-type and mutant macrophages and was not influenced by interferon gamma/lipopolysaccharide activation. Following phagocytosis of [(59)Fe]Tf-anti-Tf immune complexes, iron uptake was equivalent and up-regulated similarly with activation, but intracellular distribution was markedly different. In wild-type macrophages most iron was in the soluble (60%) rather than insoluble (12%) fraction, with 28% ferritin (Ft)-bound. With activation, the soluble component increased to 82% at the expense of Ft-bound iron (<5%). In mutant macrophages, 40 50% of iron was in insoluble form, 50-60% was soluble and <5% was Ft-bound. Western-blot analysis confirmed failure of mutant macrophages to degrade complexes 24 h after phagocytic uptake. Confocal microscopy showed that complexes were within lysosome-associated membrane protein 1-positive vesicles in wild-type and mutant macrophages at 30 min and 24 h, implying failure in the degradative process in mature phagosomes in mutant macrophages. NO-mediated iron release was 2.4-fold higher in activated wild-type macrophages compared with mutant macrophages. Overall, our data suggest that iron acquired by phagocytosis and degradation is retained within the phagosomal compartment in wild-type macrophages, and that NO triggers iron release by direct secretion of phagosomal contents rather than via the cytoplasm. PMID- 11903052 TI - Involvement of protein kinase D in Fc gamma-receptor activation of the NADPH oxidase in neutrophils. AB - Protein kinases involved in the activation of the NADPH oxidase by Fc gamma receptors in neutrophils were studied. Of three different protein kinase C (PKC) inhibitors, Go 6976 inhibited the NADPH oxidase completely, whereas bisindolylmaleimide I and Ro 31-8220 caused a 70-80% inhibition. Thus a Go 6976 sensitive, bisindolylmaleimide I/Ro 31-8220-insensitive component contributes to NADPH oxidase activation induced by Fc gamma receptors. Down-regulation of PKC isotypes resulted in inhibition of Fc gamma-receptor-activated NADPH oxidase, but a down-regulation-insensitive component was still present. This component was sensitive to Go 6976, but insensitive to Ro 31-8220. It has been shown previously that protein kinase D/PKC-mu (PKD) shows this same pharmacology in vitro. We show that PKD is present in neutrophils and that, in contrast with PKC isotypes, PKD is not down-regulated. Therefore PKD may participate in NADPH oxidase activation. To obtain direct evidence for this we adopted an antisense approach. Antisense PKD inhibited NADPH oxidase induced by Fc gamma-receptor stimulation by 50% and the Ro 31-8220-insensitive component in the activation was inhibited by antisense PKD. In vitro kinase assays showed that PKD is activated by presenting IgG opsonized particles to neutrophils. Furthermore, PKD localizes to the area of particle intake in the cell and phosphorylates two of the three cytosolic components of the NADPH oxidase, p40(phox) and p47(phox). Taken together, these data indicate that Fc gamma receptors engage PKD in the regulation of the NADPH oxidase. PMID- 11903053 TI - Major outer membrane proteins and proteolytic processing of RgpA and Kgp of Porphyromonas gingivalis W50. AB - Porphyromonas gingivalis is an anaerobic, asaccharolytic Gram-negative rod associated with chronic periodontitis. We have undertaken a proteomic study of the outer membrane of P. gingivalis strain W50 using two-dimensional gel electrophoresis and peptide mass fingerprinting. Proteins were identified by reference to the pre-release genomic sequence of P. gingivalis available from The Institute for Genomic Research. Out of 39 proteins identified, five were TonB linked outer membrane receptors, ten others were putative integral outer membrane proteins and four were putative lipoproteins. Pyroglutamate was found to be the N terminal residue of seven of the proteins, and was predicted to be the N-terminal residue of 13 additional proteins. The RgpA, Kgp and HagA polyproteins were identified as fully processed domains in outer membranes prepared in the presence of proteinase inhibitors. Several domains were found to be C-terminally truncated 16-57 residues upstream from the N-terminus of the following domain, at a residue penultimate to a lysine. This pattern of C-terminal processing was not detected in a W50 strain isogenic mutant lacking the lysine-specific proteinase Kgp. Construction of another W50 isogenic mutant lacking the arginine-specific proteinases indicated that RgpB and/or RgpA were also involved in domain processing. The C-terminal adhesin of RgpA, designated RgpA27, together with RgpB and two newly identified proteins designated P27 and P59 were found to migrate on two-dimensional gels as vertical streaks at a molecular mass 13-42 kDa higher than that calculated from their gene sequences. The electrophoretic behaviour of these proteins, together with their immunoreactivity with a monoclonal antibody that recognizes lipopolysaccharide, is consistent with a modification that could anchor the proteins to the outer membrane. PMID- 11903054 TI - Maintenance of the filamentous actin cytoskeleton is necessary for the activation of store-operated Ca2+ channels, but not other types of plasma-membrane Ca2+ channels, in rat hepatocytes. AB - The roles of the filamentous actin (F-actin) cytoskeleton and the endoplasmic reticulum (ER) in the mechanism by which store-operated Ca(2+) channels (SOCs) and other plasma-membrane Ca(2+) channels are activated in rat hepatocytes in primary culture were investigated using cytochalasin D as a probe. Inhibition of thapsigargin-induced Ca(2+) inflow by cytochalasin D depended on the concentration and time of treatment, with maximum inhibition observed with 0.1 microM cytochalasin D for 3 h. Cytochalasin D (0.1 microM for 3 h) did not inhibit the total amount of Ca(2+) released from the ER in response to thapsigargin but did alter the kinetics of Ca(2+) release. The effects of cytochalasin D (0.1 microM) on vasopressin-induced Ca(2+) inflow were similar to those on thapsigargin-induced Ca(2+) inflow, except that cytochalasin D did inhibit vasopressin-induced release of Ca(2+) from the ER. Cytochalasin D (0.1 microM) inhibited vasopressin-induced Mn(2+) inflow (predominantly through intracellular messenger-activated non-selective cation channels), but the degree of inhibition was less than that of vasopressin-induced Ca(2+) inflow (predominantly through Ca(2+)-selective SOCs). Maitotoxin- and hypotonic shock induced Ca(2+) inflow were enhanced rather than inhibited by 0.1 microM cytochalasin D. Treatment with 0.1 microM cytochalasin D substantially reduced the amount of F-actin at the cell cortex, whereas 5 microM cytochalasin D increased the total amount of F-actin and caused an irregular distribution of F actin at the cell cortex. Cytochalasin D (0.1 microM) caused no significant change in the overall arrangement of the ER [monitored using 3',3' dihexyloxacarbocyanine iodide [DiOC(6)(3)] in fixed cells] but disrupted the fine structure of the smooth ER and reduced the diffusion of DiOC(6)(3) in the ER in live hepatocytes after photobleaching. It is concluded that (i) the concentration of cytochalasin D is a critical factor in the use of this agent as a probe to disrupt the cortical F-actin cytoskeleton in rat hepatocytes, (ii) a reduction in the amount of cortical F-actin inhibits SOCs but not intracellular messenger activated non-selective cation channels, and (iii) inhibition of the activation of SOCs and reduction in the amount of cortical F-actin is associated with disruption of the organization of the ER. PMID- 11903055 TI - Protein kinase A enhances, whereas glycogen synthase kinase-3 beta inhibits, the activity of the exon 2-encoded transactivator domain of heterogeneous nuclear ribonucleoprotein D in a hierarchical fashion. AB - Heterogeneous nuclear ribonucleoprotein D (hnRNP D) is implicated in transcriptional regulation. Alternative splicing of exons 2 and 7 generates four isoforms of the protein. We report here that only isoforms that contain the product of exon 2 (amino acids 79-97) were able to transactivate. Moreover, the exon 2-encoded protein domain alone was sufficient to drive transcription. TATA binding protein and p300 interacted with a synthetic peptide corresponding to exon 2, and both proteins co-precipitated with hnRNP D. Stimulation of protein kinase A (PKA) and protein kinase C (PKC) synergistically induced the transactivating ability of hnRNP D, and the exon 2-encoded domain was sufficient for this inducibility. In kinase assays PKA phosphorylated Ser-87 of hnRNP D, whereas glycogen synthase kinase-3 beta (GSK-3 beta) phosphorylated Ser-83, but only if Ser-87 had been pre-phosphorylated by PKA. Phosphorylation of Ser-87 enhanced, whereas phosphorylation of Ser-83 repressed, transactivation. Overexpression of GSK-3 beta inhibited transactivation by hnRNP D, but stimulation of PKC negated the inhibitory effect of GSK-3 beta. We suggest that a hierarchical phosphorylation pathway regulates the transactivating ability of hnRNP D: PKA activates hnRNP D, but at the same time renders it sensitive to inhibition by GSK-3 beta; the latter inhibition can be suspended by inactivating GSK-3 beta with PKC. PMID- 11903056 TI - Glycosylation and epitope mapping of the 5T4 glycoprotein oncofoetal antigen. AB - The human 5T4 oncofoetal antigen is a focus for development of several antibody directed therapies on the basis of the murine monoclonal antibody against 5T4 (mAb5T4), which recognizes a conformational epitope. 5T4 molecules are highly N glycosylated transmembrane glycoproteins whose extracellular domain contains two regions of leucine-rich repeats (LRRs) and associated flanking regions, separated by an intervening hydrophilic sequence. Using a series of deletion and mutated cDNA constructs as well as chimaeras with the murine homologue, we have mapped the mAb5T4 epitope to the more membrane-proximal LRR2 or its flanking region. Analysis of the glycosylation of the seven consensus Asp-Xaa-Ser/Thr sites was consistent with all of the sites being glycosylated. A combination of two high mannose chains (predominantly octasaccharide) and five mostly sialylated bi-, tri and tetra-antennary complex chains with minor quantities of core fucose were detected. The two glycosylation sites, which are the most likely to have predominantly high-mannose chains, are in the only two regions that show significant differences between the human and the 81% identical mouse sequence. A site-directed mutation, which abolished glycosylation at one of these sites (position 192), did not alter antigenicity. The other, which is nearest to the N terminus in the human, has an Asn-Leu-Thr to Asn-Leu-Leu conversion in the mouse, so cannot be glycosylated in the latter species. The large complex glycosylation at the other sites is likely to influence the antigenicity and tertiary structure generating the 5T4 epitope. PMID- 11903057 TI - Comparative characterization of two DEAD-box RNA helicases in superfamily II: human translation-initiation factor 4A and hepatitis C virus non-structural protein 3 (NS3) helicase. AB - Eukaryotic initiation factor 4A (eIF4A) is an ATP-dependent RNA helicase and is homologous to the non-structural protein 3 (NS3) helicase domain encoded by hepatitis C virus (HCV). Reported here is the comparative characterization of human eIF4A and HCV NS3 helicase in an effort to better understand viral and cellular helicases of superfamily II and to assist in designing specific inhibitors against HCV infections. Both eIF4A and HCV NS3 helicase domain were expressed in bacterial cells as histidine-tagged proteins and purified to homogeneity. Purified eIF4A exhibited RNA-unwinding activity and acted on RNA or RNA/DNA but not DNA duplexes. In the absence of cellular cofactors, eIF4A operated unwinding in both the 3' to 5' and 5' to 3' directions, and was able to unwind blunt-ended RNA duplex, suggesting that bidirectionality is an intrinsic property of eIF4A. In contrast, HCV NS3 helicase showed unidirectional 3' to 5' unwinding of RNA and RNA/DNA, as well as of DNA duplexes. With respect to NTPase activity, eIF4A hydrolysed only ATP or dATP in the presence of RNAs, whereas HCV NS3 helicase could hydrolyse all ribo- and deoxyribo-NTPs in an RNA-independent manner. In parallel, only ATP or dATP could drive the unwinding activity of eIF4A whereas HCV NS3 could function with all eight standard NTPs and dNTPs. The observed differences in their substrate specificity may prove to be useful in designing specific inhibitors targeting HCV NS3 helicase but not human eIF4A. PMID- 11903058 TI - Nuclear receptor corepressor-dependent repression of peroxisome-proliferator activated receptor delta-mediated transactivation. AB - The nuclear receptor corepressor (NCoR) was isolated as a peroxisome-proliferator activated receptor (PPAR) delta interacting protein using the yeast two-hybrid system. NCoR interacted strongly with the ligand-binding domain of PPAR delta, whereas interactions with the ligand-binding domains of PPAR gamma and PPAR alpha were significantly weaker. PPAR-NCoR interactions were antagonized by ligands in the two-hybrid system, but were ligand-insensitive in in vitro pull-down assays. Interaction between PPAR delta and NCoR was unaffected by coexpression of retinoid X receptor (RXR) alpha. The PPAR delta-RXR alpha heterodimer bound to an acyl-CoA oxidase (ACO)-type peroxisome-proliferator response element recruited a glutathione S-transferase-NCoR fusion protein in a ligand-independent manner. Contrasting with most other nuclear receptors, PPAR delta was found to interact equally well with interaction domains I and II of NCoR. In transient transfection experiments, NCoR and the related silencing mediator for retinoid and thyroid hormone receptor (SMRT) were shown to exert a marked dose-dependent repression of ligand-induced PPAR delta-mediated transactivation; in addition, transactivation induced by the cAMP-elevating agent forskolin was efficiently reduced to basal levels by NCoR as well as SMRT coexpression. Our results suggest that the transactivation potential of liganded PPAR delta can be fine-tuned by interaction with NCoR and SMRT in a manner determined by the expression levels of corepressors and coactivators. PMID- 11903059 TI - Characterization of the role of the AMP-activated protein kinase in the stimulation of glucose transport in skeletal muscle cells. AB - Stimulation of AMP-activated protein kinase (AMPK) in skeletal muscle has been correlated with an increase in glucose transport. Here, we demonstrate that adenoviral-mediated expression of a constitutively active mutant of AMPK alpha leads to activation of glucose transport in a skeletal-muscle cell line, similar to that seen following treatment with 5-amino-imidazolecarboxamide (AICA) riboside, hyperosmotic stress or insulin. In contrast, expression of a dominant negative form of AMPK blocked the stimulation of glucose transport by both AICA riboside and hyperosmotic stress, but was without effect on either insulin or phorbol-ester-stimulated transport. These results demonstrate that activation of AMPK is sufficient for stimulation of glucose uptake into muscle cells, and is a necessary component of the AICA riboside- and hyperosmotic-stress-induced pathway leading to increased glucose uptake. On the other hand, AMPK is not required in the insulin- or phorbol-ester-mediated pathways. Long-term (5 days) expression of the constitutively active AMPK mutant increased protein expression of GLUT1, GLUT4 and hexokinase II, consistent with previous reports on the chronic treatment of rats with AICA riboside. Expression of constitutively active AMPK had no detectable effect on p38 mitogen-activated protein kinase levels, although interestingly the level of protein kinase B was decreased. These results demonstrate that long-term activation of AMPK is sufficient to cause increased expression of specific proteins in muscle. Our results add further support to the hypothesis that long-term activation of AMPK is involved in the adaptive response of muscle to exercise training. PMID- 11903060 TI - Opposing roles of serine/threonine kinases MEKK1 and LOK in regulating the CD28 responsive element in T-cells. AB - T-cell activation requires signals from both the T-cell receptor (TcR) and other co-stimulatory molecules such as CD28. TcR- and CD28-mediated signals are integrated during T-cell activation resulting in the expression of cytokine genes such as interleukin-2 (IL-2). An enhancer element (CD28RE) of the IL-2 gene specifically responsive to CD28 signals has been previously identified and characterized. This response element and an adjacent Activated Protein-1 (nuclear factor-interleukin-2B) site together (RE/AP1) were shown to complex with c-rel, AP-1 and other factors. However, details of the signal transduction pathways leading from CD28 to the composite response element remain poorly understood. We present data showing that overexpression of the serine threonine kinase, mitogen activated protein kinase/extracellular-signal-regulated kinase kinase kinase-1 (MEKK1), but not nuclear factor-kappa B inducing kinase, or MAP kinase/ERK kinase 1 (MEK1), can significantly increase the level of CD28RE/AP1-driven luciferase (Luc) reporter gene expression in Jurkat E6-1 cells. A MEKK1 dominant negative mutant blocked such activation induced by stimulation with Raji B cells and the superantigen staphylococcus enterotoxin E (SEE), as well as via CD3/CD28. Mutations in either site of the RE/AP1 element abolished MEKK1-induced Luc expression. Calcineurin inhibitors, CsA and FK520, or inhibitors of p38 kinase (SB 203580), or MEK1 (PD 098059), did not affect MEKK1-induced reporter activation. These results directly implicate MEKK1 in the CD28 signalling pathway that activates the CD28 response element. Co-expression of the lymphocyte oriented kinase (LOK) kinase attenuated Raji/SEE-induced IL-2 production in Jurkat cells, as well as MEKK1 and Raji/SEE-induced reporter gene activation. These data suggest that MEKK1 and LOK may have opposing roles in regulating the CD28RE/AP1 element. PMID- 11903061 TI - De novo-synthesized ceramide is involved in cannabinoid-induced apoptosis. AB - Delta(9)-Tetrahydrocannabinol (THC) and other cannabinoids have been shown to induce apoptosis of glioma cells via ceramide generation. In the present study, we investigated the metabolic origin of the ceramide responsible for this cannabinoid-induced apoptosis by using two subclones of C6 glioma cells: C6.9, which is sensitive to THC-induced apoptosis; and C6.4, which is resistant to THC induced apoptosis. Pharmacological inhibition of ceramide synthesis de novo, but not of neutral and acid sphingomyelinases, prevented THC-induced apoptosis in C6.9 cells. The activity of serine palmitoyltransferase (SPT), which catalyses the rate-limiting step of ceramide synthesis de novo, was remarkably enhanced by THC in C6.9 cells, but not in C6.4 cells. However, no major changes in SPT mRNA and protein levels were evident. Changes in SPT activity paralleled changes in ceramide levels. Pharmacological inhibition of ceramide synthesis de novo also prevented the stimulation of extracellular-signal-regulated kinase and the inhibition of protein kinase B triggered by cannabinoids. These findings show that de novo-synthesized ceramide is involved in cannabinoid-induced apoptosis of glioma cells. PMID- 11903062 TI - Glutamic acid-65 is an essential residue for catalysis in Proteus mirabilis glutathione S-transferase B1-1. AB - The functional role of three conserved amino acid residues in Proteus mirabilis glutathione S-transferase B1-1 (PmGST B1-1) has been investigated by site directed mutagenesis. Crystallographic analyses indicated that Glu(65), Ser(103) and Glu(104) are in hydrogen-bonding distance of the N-terminal amino group of the gamma-glutamyl moiety of the co-substrate, GSH. Glu(65) was mutated to either aspartic acid or leucine, and Ser(103) and Glu(104) were both mutated to alanine. Glu(65) mutants (Glu(65)-->Asp and Glu(65)-->Leu) lost all enzyme activity, and a drastic decrease in catalytic efficiency was observed for Ser(103)-->Ala and Glu(104)-->Ala mutants toward both 1-chloro-2,4-dinitrobenzene and GSH. On the other hand, all mutants displayed similar intrinsic fluorescence, CD spectra and thermal stability, indicating that the mutations did not affect the structural integrity of the enzyme. Taken together, these results indicate that Ser(103) and Glu(104) are significantly involved in the interaction with GSH at the active site of PmGST B1-1, whereas Glu(65) is crucial for catalysis. PMID- 11903064 TI - Phlogiston--fire air--oxygen. The fascinating story of an 18th century discovery. PMID- 11903063 TI - Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. AB - Dysregulation of the human transforming acidic coiled-coil (TACC) proteins is thought to be important in the evolution of breast cancer and multiple myeloma. However, the exact role of these proteins in the oncogenic process is currently unknown. Using the full-length TACC1 protein as bait to screen a human mammary epithelial cDNA library, we have identified two genes that are also amplified and overexpressed in tumours derived from different cellular origins. TACC1 interacts with the C-terminus of both the microtubule-associated colonic and hepatic tumour overexpressed (ch-TOG) protein, and the oncogenic transcription factor glioma amplified sequence 41/NuMA binding protein 1 (GAS41/NuBI1; where NuMA stands for nuclear mitotic apparatus protein 1). This suggests that the TACC proteins can form multiple complexes, dysregulation of which may be an important step during tumorigenesis. PMID- 11903065 TI - Priestley, the furious free thinker of the enlightenment, and Scheele, the taciturn apothecary of Uppsala. PMID- 11903067 TI - Effective inhibition of nitric oxide production by aminoguanidine does not reverse hypotension in endotoxaemic rats. AB - BACKGROUND: Excess production of nitric oxide (NO) by the inducible NO synthase (iNOS) has been implicated in the pathophysiology of septic shock. Using methaemoglobin (metHb) and the stable NO metabolite nitrate as markers of NO formation, we assessed the effect of iNOS blockade by aminoguanidine (AG) on hypotension and NO formation in endotoxaemic rats. METHODS: In 32 male Wistar rats under chloralose anaesthesia, MetHb (at 15 and 330 min, respectively) and plasma nitrate (at 330 min) were determined. Mean arterial pressure, heart rate and haematocrit were monitored. The LPS group (n=8) received bacterial endotoxin (LPS), 3 mg kg(-1) i.v. and was subsequently monitored for 5 h. At 2 h after LPS, the LPS+AG20 group (n=8) received AG, 5 mg kg(-1), and 5 mg kg(-1) h(-1) for the remaining 3 h. The LPS+AG100 group (n=8) instead received 25 mg kg(-1), followed by 25 mg kg(-1) h(-1). The NaCl group (n=8) was given corresponding volumes of isotonic saline. RESULTS: AG decreased the LPS-induced rise in plasma nitrate by about 50% in the LPS+AG20 group. MetHb levels, however, were not appreciably reduced by this dose. Both NO metabolites reached control levels after the higher dose of AG. LPS caused a progressive decrease in haematocrit. AG did not influence the LPS-induced hypotension, tachycardia or haemodilution. CONCLUSION: AG inhibited NO formation in a dose-dependent way. Yet, AG had no haemodynamic effects, suggesting a minor cardiovascular influence of iNOS in this endotoxin model, in parallel to what has been found in microbial sepsis. PMID- 11903066 TI - Changes in serum S100beta protein and Mini-Mental State Examination after cold (28 degrees C) and warm (34 degrees C) cardiopulmonary bypass using different blood gas strategies (alpha-stat and pH-stat). AB - BACKGROUND: The effect of cardiopulmonary bypass temperature and blood gas management on the brain is still controversial. This study was designed to compare the changes in S100beta protein concentration and Mini-Mental State Examination in patients undergoing cold (28 degrees C) vs. warm (34 degrees C) cardiopulmonary bypass using different blood gas strategies (alpha-stat and pH stat). METHODS: Sixty patients were randomly allocated to one of four equal groups (cold alpha-stat, cold pH-stat, warm alpha-stat, warm pH-stat). Serum S100beta concentrations were measured before CPB, directly after CPB, at 4.5 h and at 24 h after CPB. Mini-Mental State Examination was performed one day before surgery and on day five after the operation. Antegrade warm blood cardioplegia (37 degrees C) was used in all patients. RESULTS: There was no significant difference in postoperative S100beta protein levels between the four groups. Also, there was no interaction between bypass temperature and type of blood gas strategy on S100beta levels after bypass (directly after bypass, 4.5 h and 24 h after bypass). Mini-Mental State Examination score was not affected by blood gas strategy but it was significantly lower in patients undergoing cold cardiopulmonary bypass surgery: median (range), 26 (12-29) vs. 27 (23-30) in warm patients, P = 0.014. There was no significant correlation between Mini-Mental State Examination score 5 days after CPB and S100beta levels at any of the studied time-points after CPB. CONCLUSION: These results support the use of warm CPB (34 degrees C) in patients undergoing coronary artery bypass surgery regardless of the type of blood gas strategy. PMID- 11903069 TI - The influence of anaesthesia and surgery on the circadian rhythm of melatonin. AB - BACKGROUND: Operations are typically associated with sleep and other circadian rhythm disturbances. The present study was set up to evaluate the influence of spinal and general anaesthesia associated with knee surgery on the circadian rhythm of melatonin, which has sleep inducing properties. Previously this context has been studied only in some invasive operations and it might be that general anaesthesia induces more disturbances on circadian rhythm of melatonin than operations done with patients awake. METHODS: The circadian secretion pattern of melatonin was monitored during the pre- and postoperative evenings, nights and mornings to clarify possible anaesthesia/surgery-induced changes in the nocturnal secretion of melatonin and in the phase of the melatonin rhythm. The study included 20 patients scheduled for minor orthopaedic operations. The patients were randomised to receive either spinal or general anaesthesia. Melatonin was measured from evening and morning saliva samples radioimmunologically. The nocturnal urine before and after surgery was radioimmunologically examined for 6 hydroxymelatonin sulphate. RESULTS: Melatonin secretion evaluated from the saliva samples was significantly diminished during the first postoperative evening as compared with that during the preoperative evening (P<0.001). There was also a significant decline of 26% (P<0.05) in postoperative 6-hydroxymelatonin sulphate excretion. There was no significant difference in melatonin secretion between the spinal and general anaesthesia groups. CONCLUSION: Our findings suggest that anaesthesia in conjunction with surgery acutely disturbed the normal circadian rhythm of melatonin by delaying the onset of nocturnal melatonin secretion. PMID- 11903068 TI - Effects of nicorandil on myocardial function and metabolism in the post-ischaemic reperfused heart with or without inhalation anaesthetics. AB - BACKGROUND: Nicorandil, which is an ATP-sensitive K channel opener, has been reported to protect the ischaemic myocardium. However, its interaction with inhalation anaesthetics on the ischaemic myocardium has not been well elucidated. So, we have investigated whether isoflurane or sevoflurane modify the effects of nicorandil on cardiac function and metabolism in the rat heart-lung preparation. METHODS: Animals were allocated to 4 groups as follows: Control group, no drug; Nic group, nicorandil; Nic+Iso group, nicorandil and isoflurane; Nic+Sev group, nicorandil and sevoflurane. Seven minutes after the start of perfusion, nicorandil was administered and 10 min after the start of perfusion, the heart was rendered globally ischaemic for 10 min, and then the heart was reperfused for 10 min. RESULTS: LVdP/dt max in the Nic group was higher than those in the other groups. Right atrial pressure in the Nic+Iso and Nic+Sev groups was significantly higher than in the Control and Nic groups. Myocardial ATP in the Nic group was higher than in the other groups. DHBA levels in the perfusate in the Nic and Nic+Iso groups were lower than those in the Control and Nic+Sev groups, but those in the Nic+Sev group were higher than those in the other groups. CONCLUSIONS: Nicorandil improved post-ischaemic cardiac function and preserved high-energy phosphates. However, these beneficial effects of nicorandil were abolished by the combination with isoflurane or sevoflurane. In addition, sevoflurane increased hydroxyl radical formation in the post-ischaemic reperfused heart. PMID- 11903070 TI - Reducing the risk of systemic embolization during gynecologic laparoscopy--effect of volume preload. AB - BACKGROUND: About 27% of the population is known to have a patent foramen ovale. It can be opened if the left atrial pressure is less than the right atrial pressure. This pressure reversal has been reported during gynecologic laparoscopic surgery. The present paper describes with help of transesophageal echocardiography the pressure relationship between the atria during laparoscopic surgery and the effect of volume preload. METHODS: Twenty-one gynecologic ASA 1-3 patients were included in this open study. The movement of interatrial septum was monitored with transesophageal echocardiography during the procedure. If the septum movement was to the left, the patient was given 500 mL hydroxyethyl starch to increase the filling pressures. RESULTS: After induction, the mobile part of foramen ovale rounded to the right in 15 patients but six patients showed movement to the left. After pneumoperitoneum and head-down tilt, one patient of the six returned to normal but eight additional patients showed movement to the left. These 13 patients had a filling infusion of 500 mL hydroxyethyl starch. The movement was normalized in 12 patients. We saw echogenic particles coming from the inferior caval vein in every patient. Only one patient had very small atrial septal defects and no embolic complications. CONCLUSION: The head-down tilt and pneumoperitoneum had a more negative influence on the filling of the left side than on the filling of the right side of the heart. The pressure reversal occurs in systole during expiration of mechanical ventilation. The infusion of volume helps to normalize the pressure relationship and to diminish the embolic risk. PMID- 11903071 TI - Construction and evaluation of a manikin for perioperative heat exchange. AB - BACKGROUND: During surgery hypothermia can be avoided only if the heat exchange between the body surface and the environment can be controlled. To allow a systematic analysis of this heat exchange, we constructed and evaluated a copper manikin of the human body. METHODS: The manikin consists of six tubes (head, trunk, two arms and two legs) painted matt-black to simulate the emissivity of the human skin. Hot-water mattresses are bonded to the inner surface of the copper tubes to set the surface temperature. Calibrated heat flux transducers were placed on the following points to determine the heat exchange coefficient for radiation and convection (hRC) of the manikin: Forehead, chest, abdomen, upper arm, forearm, dorsal hand, anterior thigh, anterior leg and foot. Room temperature was set to 22 degrees C. Surface temperature of the manikin was set between 22 degrees C and 38 degrees C. The hRC was determined by linear regression analysis as the slope of the temperature gradient between the manikin and the room versus the measured heat flux. Subsequently we studied five minimally clothed volunteers in a climate chamber. Initial chamber temperature was set to 29 degrees C and was lowered slowly to 12 degrees C. The hRC was determined as described above for each volunteer. RESULTS: The hRC of the manikin was 11.0 W m(-2) degrees C(-1) and hRC of the volunteers was 10.8 W m(-2) degrees C(-1). CONCLUSION: The excellent correlation of hRC between the volunteers and the manikin will allow the manikin to be used for standardised studies of perioperative heat exchange. PMID- 11903072 TI - Temperature-related fluid extravasation during cardiopulmonary bypass: an analysis of filtration coefficients and transcapillary pressures. AB - BACKGROUND: Cardiopulmonary bypass (CPB) as used for cardiac surgery and for rewarming individuals suffering deep accidental hypothermia is held responsible for changes in microvascular fluid exchange often leading to edema and organ dysfunction. The purpose of this work is to improve our understanding of fluid pathophysiology and to explore the implications of the changes in determinants of transcapillary fluid exchange during CPB with and without hypothermia. This investigation might give indications on where to focus attention to reduce fluid extravasation during CPB. METHODS: Published data on "Starling variables" as well as reported changes in fluid extravasation, tissue fluid contents and lymph flow were analyzed together with assumed/estimated values for variables not measured. The analysis was based on the Starling hypothesis where the transcapillary fluid filtration rate is given by: JV=Kf [Pc-Pi-sigma(COPp-COPi)]. Here Kf is the capillary filtration coefficient, sigma the reflection coefficient, P and COP are hydrostatic and colloid osmotic pressures, and subscript 'c' refers to capillary, 'i' to the interstitium and 'p' to plasma. RESULTS AND CONCLUSION: The analysis indicates that attempts to limit fluid extravasation during normothermic CPB should address primarily changes in Kf, while changes in both Kf and Pc must be considered during hypothermic CPB. PMID- 11903073 TI - Splanchnic vasoconstriction by angiotensin II is arterial pressure dependent. AB - BACKGROUND: Our hypothesis was that splanchnic vasoconstriction by exogenous angiotensin II (Ang II) is significantly potentiated by local mechanisms increasing vasomotor tone and that splanchnic tissue oxygenation during administration of Ang II is perfusion pressure dependent. The aim was to study local splanchnic circulatory effects and tissue oxygenation during intravenous infusion of Ang II at different levels of regional arterial driving pressure in a whole-body large animal model. METHODS: Ang II was infused in incremental doses (0-200 microg x h-1) in anaesthetised instrumented pigs (n=8). Mean superior mesenteric arterial pressure (PSMA) was adjusted by a local variable perivascular occluder. Perivascular ultrasound and laser-Doppler flowmetry were used for measurements of mesenteric venous blood flow and superficial intestinal blood flow, respectively. Intestinal oxygenation was evaluated by oxygen tissue tension (PtiO2) and lactate fluxes. RESULTS: Ang II produced prominent and dose-dependent increases in mesenteric vascular resistance (RSMA) when the intestine was exposed to systemic arterial pressure, but Ang II increased RSMA only minimally when PSMA was artificially kept constant at a lower level (50 mmHg) by the occluder. Although Ang II decreased PtiO2 at a PSMA of 50 mmHg, splanchnic lactate production was not observed. CONCLUSION: We demonstrate that splanchnic vasoconstriction by exogenous Ang II is dependent on arterial driving pressure, suggesting significant potentiation through autoregulatory increases in vasomotor tone. Intestinal hypoxaemia does not seem to occur during short-term infusion of Ang II in doses that significantly increases systemic arterial pressure. PMID- 11903074 TI - Effect of amino acid solutions on intraoperative hypothermia and postoperative shivering. Comparison of two anesthetic regimens. AB - BACKGROUND: Intraoperative hypothermia is a major adverse effect of general anesthesia. The different anesthetics may influence thermoregulation differently. Proteins or amino acids have been postulated to stimulate heat production. The purpose of this study is to compare the effects of intraoperative administration of amino acid solutions on intraoperative hypothermia and postoperative shivering in two different anesthetic regimens. METHODS: Forty ASA I-III craniotomy patients were assigned to four groups of 10 patients in a randomized prospective study, as follows: ISO - isoflurane-based anesthesia; PRO - propofol-based anesthesia; ISO + AA - Isoflurane-based anesthesia with supplementation of amino acid infusion; PRO + AA - Isoflurane-based anesthesia with supplementation of amino acid infusion. Hemodynamic parameters, esophageal temperature and postoperative shivering scores were recorded. RESULTS: Core temperatures were higher during emergence in amino acid-treated propofol group, compared with the other groups. The core temperature decreased significantly in three groups throughout the operation, except the in amino acid-treated propofol group. The shivering intensity was less in the amino acid-treated groups. CONCLUSION: The anesthetic method may influence the thermic effect of amino acids under general anesthesia. Propofol anesthesia has more thermogenic effect than isoflurane when combined with amino acid solutions. PMID- 11903075 TI - The effects of platelet apheresis in total hip replacement surgery on platelet activation. AB - BACKGROUND: Autologous platelet rich plasma (PRP) harvest with autotransfusion devices has been used for 10 years in cardiac surgery and recently in orthopedics as a blood saving method. The quality of the harvested platelets has not been adequately examined, in part because of methodological difficulties in studying platelet function during surgery. METHODS: Twenty patients undergoing primary total hip replacement (THR) were studied. Ten patients underwent an immediate preoperative platelet apheresis to obtain concentrated platelet rich plasma (c PRP). The other 10 patients not undergoing apheresis were allocated to a control group. Platelet activation was evaluated as the population expressing P-selectin on the surface of platelets in the c-PRP and in blood samples collected pre-, per and postoperatively. The method used was flow cytometry. RESULTS AND CONCLUSIONS: A minor population of activated platelets was found to be circulating in the patients' blood, with a highly significant difference between patients (P = 0.005), and with a range of 1-23% in peroperative activation. PRP harvest did not significantly alter platelet activity. The platelet apheresis procedure did not inhibit platelet function in the c-PRP, as judged by a high proportion of platelets that could be activated in ADP stimulation experiments (mean value +/- SD 86% +/- 7.5%). PMID- 11903076 TI - Pigs are not a reliable experimental model in the study of the haemodynamic and respiratory effects of CO2 pneumoperitoneum. AB - BACKGROUND: Haemodynamic and respiratory effects of a CO2 pneumoperitoneum (intra abdominal pressure = 12 mmHg) associated to a head-up position(15 degrees ) were studied in 20 pigs using a Swan-Ganz catheter and the Single Breath Test for CO2. The pneumoperitoneum induced a moderate rise in mean arterial pressure (+17%) (P<0.001) without any variation in heart rate, cardiac output and systemic vascular resistances. RESULTS: The following respiratory effects were observed: an increase in PaCO2 (+20%) (P<0.001), PE'CO2 (+31%) (P<0.001), expired volume of CO2 (+28%) (P<0.001), arterial to end-tidal CO2 gradient (+80%) (P<0.001) and alveolar dead space (+40%) (P<0.001) occured. Alveolar ventilation remained stable. Finally and contrary to healthy human patient, intraperitoneal CO2 insufflation in pig induced slight haemodynamic changes and major respiratory modifications. CONCLUSION: Thus, our data do not support the conclusion that the pig is a reliable experimental model for studying the pathophysiology of CO2 pneumoperitoneum-induced changes in haemodynamic and respiratory parameters, in human patients. PMID- 11903077 TI - Intravenous fluids warming by mattress is simple and efficient during pediatric surgery. AB - BACKGROUND/AIM: Temperature control is essential during pediatric surgery. The effectiveness of two methods of warming intravenous (i.v.) fluids to preserve normothermia was compared during abdominal surgery. METHODS: Intraoperative core temperature (CT) was measured in 59 children, 8 years of age and younger. Patients were randomly allocated to two study groups according to the method of fluids warming. In Group M (n = 30), fluids were warmed by placing the i.v. tubing under a warming mattress, and in Group T (n = 29), by using an active i.v. fluid tube warming system. Observations of CT, infusion fluid temperature (IFT) at the entry to the patient and other relevant parameters were made at 30-min intervals throughout the surgical procedure. RESULTS: The two groups did not differ significantly by age, gender, body weight or length of surgical procedure. Although baseline IFT was significantly lower in Group M than in Group T (33.8 degrees C vs. 35.1 degrees C), it increased during the procedure by 1.2 degrees C in Group M compared to a 0.2 degrees C increase in Group T. Baseline CT was also lower by 0.5 degrees C in Group M compared to Group T (NS), but CT increased in Group M by 1.0 degrees C compared to 0.2 degrees C in Group T, resulting in similar final CTs. CONCLUSIONS: The increase in CT was more pronounced in patients where fluids were warmed under the warming mattress (Group M) than in those with fluids warmed by a coil warming device (Group T). The elevation in CT seen in Group M is associated with an increase in infusion fluid temperature at the line just before the i.v. cannula. Both methods of fluid warming (by placing the i.v. tubing under warming mattress and by using a fluid warming system) effectively preserved normothermia during abdominal surgery in children. PMID- 11903079 TI - Intrathecal administration of liposomal neostigmine prolongs analgesia in mice. AB - BACKGROUND: There is substantial evidence that cholinomimetic drugs increase pain threshold. However, the profound side effects of these agents have limited their clinical use either as analgesics or as analgesic adjuncts. A delivery system that would assure a slow and sustained drug release may be of value in ameliorating the problem of untoward effects. METHODS: The acetylcholinesterase inhibitor neostigmine was encapsulated into multilamellar lipid vesicles composed of phosphocholine and cholesterol. Three doses of plain or liposomal neostigmine were administered by the intrathecal route to mice (n=8-10/group), and analgesic duration was quantified by tail flick test. The doses were chosen based on preliminary experiments, which showed the maximum tolerated intrathecal doses of plain and liposomal neostigmine formulation were 0.625 microg and 80 microg, respectively. Two other doses for each formulation were then derived by 1:1 serial dilutions. Results were compared using survival analysis. RESULTS: The median antinociceptive duration for plain neostigmine was 0.33, 0.99 and 1.0 h for the 0.115, 0.312 and 0.625 microg doses, respectively. For the liposomal formulation, the median antinociceptive duration was 1.0, 1.5 and 6.0 h for the 20, 40 and 80 microg doses, respectively. CONCLUSIONS: Liposomal neostigmine provides prolonged spinal antinociception, and permits the safe administration of a relatively large dose, because drug is gradually released from the liposomal depot. This technology holds promise for the development of a clinically useful neostigmine formulation to provide spinal analgesia. PMID- 11903081 TI - The neck crease as a landmark of Chassaignac's tubercle in stellate ganglion block: anatomical and radiological evaluation. AB - BACKGROUND: Stellate ganglion block (SGB) is most commonly performed at the transverse process of the sixth cervical vertebra, the identification of which could be difficult in patients with short and wide necks. This study was conducted to evaluate whether the neck skin crease is a reliable indicator of the C6 level. METHODS: Forty-nine relatively obese pain clinic patients were investigated. They assumed a standard position for SGB. A radiopaque wire was placed along the neck skin crease caudad to the thyroid cartilage. Next, a radiopaque indicator was placed on the skin above the tubercle found to be most prominent by palpation. X-rays of the neck were obtained after each procedure. RESULTS: The probability that the neck crease would cross C5, C6 and C7 was 16%, 71%, and 12%, respectively. The most prominent tubercle corresponded to the C5, C6 and C7 levels in 16%, 69% and l4% of cases, respectively. CONCLUSION: The studied means to identify the C6 transverse process was found to correlate well with each other (P<0.001). Since in 30% of cases the C6 process could not be identified by any of the studied means, radiological guidance is recommended in order to ensure optimal safety and efficacy of SGB in selected cases. PMID- 11903078 TI - Cost-effectiveness of analgesia after Caesarean section. A comparison of intrathecal morphine and epidural PCA. AB - BACKGROUND: Patient-controlled analgesia (PCA) techniques and intrathecal morphine are the most widely used treatments for post-Caesarean section pain. However these methods have not been compared with respect to analgesic quality and cost differences. METHODS: Fifty-three patients scheduled for elective or semi-urgent Caesarean section were randomized to receive for postoperative analgesia either epidural PCA with a mixture containing bupivacaine 0.06% and sufentanil 1 microg x ml(-1) or intrathecal morphine 0.15 mg together with the spinal anaesthetic and to be supplemented with paracetamol and tramadol. Analgesic efficacy, side-effects and costs were calculated during 48 h. RESULTS: VAS pain scores both at rest and during mobilization were lower in the PCA group, more particularly during the second postoperative day. Nausea and vomiting were more frequently registered in the morphine treated patients. PCA treated patients stayed longer in the recovery room but required fewer nurse interventions on the surgical ward. Manpower and drug costs were equal in both groups. The differences in total costs (Euro) amounted to euros 33 and were mainly caused by the more expensive equipment required for epidural PCA. Satisfaction and hospital discharge were similar for both treatments. CONCLUSIONS: It was concluded that epidural PCA induced better pain relief, caused less nausea/vomiting but was more expensive than intrathecal morphine. PMID- 11903080 TI - Post-operative analgesia following total knee replacement: an evaluation of the addition of an obturator nerve block to combined femoral and sciatic nerve block. AB - BACKGROUND: Femoral and sciatic nerve block may not provide complete post operative analgesia following total knee replacement. This study was designed to evaluate whether the addition of an obturator nerve block to combined femoral and sciatic nerve block improves the quality of post-operative analgesia following primary total knee replacement. METHODS: Sixty patients were randomised into one of two groups: combined femoral and sciatic nerve block with 15 ml 0.75% ropivacaine to each nerve or combined femoral and sciatic nerve block with 15 ml 0.75% ropivacaine to each nerve and an obturator nerve block with 5 ml 0.75% ropivacaine. RESULTS: Peripheral nerve blocks were successful in 85% of patients. The group which received the obturator nerve block showed a significant increase in the time until their first request for analgesia (mean 257.0 vs. 433.6 min) and a significant reduction in the total requirements for morphine throughout the study period (mean 83.8 vs. 63.0 mg) (P<0.05). There were no systemic or neurological sequelae in any of the groups. CONCLUSIONS: The addition of an obturator nerve block to femoral and sciatic blockade improved post-operative analgesia following total knee replacement. PMID- 11903082 TI - The effect of sevoflurane on glutamate release and uptake in rat cerebrocortical presynaptic terminals. AB - BACKGROUND: Volatile anaesthetics exert their effect in the brain mainly by reducing synaptic excitability. Isoflurane abates excitation by reducing the release and increasing the uptake of transmitter glutamate into the presynaptic terminal. The exact molecular mechanisms exerting these effects, however, are not clear. Voltage-gated calcium channels have been proposed as the pharmacological target. The present study examines the effect of sevoflurane on synaptic glutamate release and free cytosolic calcium and the effect on high- and low affinity uptake of L-glutamate using isolated presynaptic terminals prepared from rat cerebral cortex. METHODS: Released glutamate was measured fluorometrically in a spectrophotometer as the fluorescence of NADPH and calcium as the fluorescence of fura-2. 4-aminopyridine was used to induce membrane depolarization. Glutamate uptake was measured in a series of different concentrations of L-glutamate corresponding to the high- and the low- affinity uptake systems adding a fixed concentration og radiolabelled glutamate. The labelling was measured by counting disintegrations per min in a beta-scintillation counter. RESULTS: Sevoflurane reduced the calcium-dependent glutamate release in a dose-dependent manner as sevoflurane 1.5, 2.5 and 4.0% reduced the release by 58, 69 and 94%, respectively (P<0.05). Membrane depolarization induced an increase in free cytosolic calcium by 25%. Sevoflurane did not affect this increase. Neither the high- nor the low affinity uptake transporter systems are affected by the anaesthetic. CONCLUSION: These results indicate that different volatile anaesthetics may act differently on the presynaptic terminal. The exact modes of action have to be further investigated. PMID- 11903083 TI - Incidence and severity of postoperative nausea and vomiting are similar after metoclopramide 20 mg and ondansetron 8 mg given by the end of laparoscopic cholecystectomies. AB - BACKGROUND: Ondansetron has a well documented antiemetic prophylactic effect, whereas in most studies of postoperative nausea and vomiting (PONV), metoclopramide is less efficacious. This can be attributed to the short-lasting effect of metoclopramide when a low dose is given at the beginning of surgery. We wanted to test a 20-mg dose of metoclopramide given at the end of surgery, using ondansetron 8 mg as a reference. METHODS: 122 patients scheduled for elective laparoscopic cholecystectomy under general anesthesia were studied in a randomized, double-blind study design. At the end of the procedure, the patients received either metoclopramide 20 mg or ondansetron 8 mg intravenously. The patients were observed for 24 h for PONV, pain, side-effects and need for rescue antiemetic medication. RESULTS: No significant differences in the incidence of PONV or need for rescue antiemetic treatment was observed in the 0-24 h postoperative study period. The overall incidence of PONV was 43% in the ondansetron group and 47% in the metoclopramide group. The ondansetron patients had a significantly higher incidence of moderate or strong pain during the postoperative observation period (61% vs. 35% in the metoclopramide group) (P < 0.05). No significant differences in side-effects between the groups were observed. CONCLUSIONS: Metoclopramide 20 mg i.v. given at the end of laparoscopic cholecystectomy resulted in a similar incidence of PONV compared with ondansetron 8 mg. The patients receiving metoclopramide had less pain than the patients receiving ondansetron. PMID- 11903084 TI - Successful treatment of restless legs syndrome with an implanted pump for intrathecal drug delivery. AB - Two patients with incapacitating symptoms from restless legs syndrome, not adequately responding to conventional treatment with dopaminergic drugs, were implanted with a pump device (Isomed) for intrathecal delivery of morphine and bupivacaine. The treatment resulted in total resolution of all symptoms with few side effects. PMID- 11903086 TI - Normal pressure hydrocephalus and cerebral blood flow: a review. AB - Normal pressure hydrocephalus is a neurological disease which poses both diagnostic and therapeutic problems for the clinician. The measurement and characterisation of cerebral blood flow has been proposed as a tool for resolving such problems as well as elucidating its pathophysiology. We review the results of studies in which this tool has been applied to normal pressure hydrocephalus patients and consider the merits of the techniques that have been utilised. Finally, consideration is given to feasible future studies and the methods that could be employed in the study of cerebral blood flow and metabolism in patients with normal pressure hydrocephalus. PMID- 11903087 TI - Duodenal levodopa infusion in Parkinson's disease--long-term experience. AB - Motor fluctuations in parkinsonian patients can be reduced by intraduodenal infusion of levodopa. Between 1991 and 1998 continuous daytime administration of levodopa through a transabdominal port has been used in 28 very advanced patients over a total period of 1045 months. A stable suspension of levodopa and carbidopa (Duodopa) has been developed. Patients were characterized by early onset, long history of disease and levodopa therapy. The reason for infusion was in all cases related to on-off fluctuations. All patients experienced a general improvement after the introduction of continuous treatment. There have been no severe complications. Six patients have taken the decision to curtail their treatment. The mean daily levodopa consumption has been slightly reduced on infusion as compared to oral therapy. Nine of the first group of patients participating in the new therapy have been regularly evaluated by means of rating scales and movement analyses. Short-term results have already been published and a follow-up showing continued positive effect after 4-7 years of continuous duodenal infusion is presented. PMID- 11903088 TI - Allergy and childhood epilepsy: a close relationship? AB - OBJECTIVES: The possibility that certain foods or allergens may induce convulsions has already been reported in the literature. None of the relevant studies has, however, shown a close correlation between allergy and epilepsy, most reports being anecdotal and open to various causal hypotheses. The case control study reported here was undertaken to test the hypothesis that epilepsy is linked to allergy. MATERIAL AND METHODS: Seventy-two epileptic children and a group of 202 controls in the same age bracket were investigated for allergy together with their immediate families. RESULTS: The study revealed significantly higher rates of eczema in the mothers and rhinitis in the siblings of the patients studied as well as generally higher incidence of allergic pathologies in both of these groups with respect to the relevant controls. A significantly higher incidence of allergy to cow's milk and asthma was also documented in the epileptic children with respect to the control group. Prick tests gave a significantly higher rate of positive results for cow's milk proteins in the cases examined with respect to the controls. The total serum IgE of a random sample of cases and controls showed no difference in mean values. CONCLUSION: The study appears to bear out the hypothesis of a higher incidence of allergy in the children with epilepsy and their immediate families than in the controls and their families. PMID- 11903089 TI - Prevalence of multiple sclerosis in Belgrade, Yugoslavia. AB - OBJECTIVES: To estimate the distribution of multiple sclerosis in the Belgrade population. METHODS: All persons who were affected and/or died from multiple sclerosis (Poser's criteria), with residence in the Belgrade region had been collected from January 1, 1985 to December 31, 1996. Prevalence was adjusted by direct method, using world population. RESULTS: From 1985 to 1996, 823 patients were suffering from multiple sclerosis. Sex ratio was 1:1.9. The mean age at onset was 32.2 +/- 9.8 years. A relapsing-remitting course of multiple sclerosis was reported in 50.7% patients, secondary progressive in 36.4%, patients, and primary progressive in 12.9% patients. On December 31, 1996, age-adjusted prevalence of multiple sclerosis in Belgrade was 41.5/100,000, 28.2/100,000 for males, and 54.1/100,000 for females. During the period studied, statistically highly significant increasing trend of multiple sclerosis prevalence was observed (P = 0.0001). CONCLUSIONS: According to findings presented in this study, Belgrade is an area with high prevalence of multiple sclerosis. PMID- 11903090 TI - Blood pressure variability and leukoaraiosis amount in cerebral small-vessel disease. AB - OBJECTIVES: To analyze the correlation between blood pressure (BP) variability and leukoaraiosis (LA) amount in patients with symptomatic cerebral small-vessel disease. MATERIALS AND METHODS: We included 25 hypertensive patients: 13 with Binswanger's disease (BD) and 12 with a first-ever lacunar infarction (LI). Baseline office BP was obtained for 3 consecutive weeks. From a 24-h ambulatory BP monitoring performed 1 week later we obtained average systolic (SBP) and diastolic (DBP) BP for daytime, nighttime and 24-h periods. SBP and DBP variability was defined as the within-subject standard deviation of all readings. A standardized cerebral MR was performed in each patient and an LA score was calculated. RESULTS: No statistically significant correlation was obtained between the LA score and any of the following BP values: 1) Baseline SBP and DBP; 2) 24-h, daytime or nighttime SBP and DBP, and 3) 24-h, daytime or nighttime SBP and DBP variability. CONCLUSION: Increased BP variability is not associated with greater amounts of leukoaraiosis. PMID- 11903091 TI - Patients in a persistent vegetative state: caregiver attitudes and reactions. AB - OBJECTIVES: This exploratory study investigated the problems encountered by caregivers of long-stay hospital patients in a persistent vegetative state. MATERIAL AND METHODS: Sixteen primary caregivers completed questionnaires designed to assess their personality, psychophysical distress, coping strategies and caregiving-related problems. RESULTS: Males showed a higher level of emotional distress and neuroticism than females. All of the caregivers used situation-oriented coping strategies less over time. had apparently unsatisfactory family relationships, and their emotional distress increased with disease duration. The thoughts of the possible death of the patient were associated with anxiety and depressive symptoms. The caregivers' everyday lives were characterized by limited social relationships, and indoor and outdoor interests. CONCLUSION: Our study underlines the importance of psychosocially assessing PVS patient caregivers, who are often alone in coping with a irreversible situation. It also introduces a questionnaire (FSQ2) that seems to be sufficient to assess the caregiving-related problems. PMID- 11903092 TI - Abnormal exploratory eye movements in schizophrenic patients vs healthy subjects. AB - OBJECTIVE: We examined previously described exploratory eye movements abnormalities as biologic markers in schizophrenic patients in comparison with age-matched healthy subjects. MATERIAL AND METHODS: Using an eye-mark recorder, eye movements were analysed for mean gazing time, total number of gazing points, mean eye scanning length, total eye scanning length, and total gazing times as subjects viewed six simple pictures in preparation for copying them. RESULTS: In patients, and to a lesser extent, out-patients, with schizophrenia showed a longer gazing time, fewer gazing points, a shorter mean and total eye scanning length. and longer gazing time than healthy subjects. In schizophrenic patients. negative symptom scores were positively correlated with mean gazing time (r = 0.33), and negatively correlated with the total number of gazing points (r = 0.29) as well as, the mean (r = -0.40) and total scanning length (r = -0.46). CONCLUSION: Exploratory eye movements are a biologic marker useful for evaluation of schizophrenia. PMID- 11903093 TI - Familial carpal tunnel syndrome: a report of a Finnish family. AB - The existence of familial carpal tunnel syndrome (FCTS) as a separate autonomic entity has been discussed during the last few years. In order to contribute with more data to the literature, we report here the results of clinical. electrophysiological, pathological and radiological studies performed in 5 patients belonging to the same Finnish pedigree. The disease appeared usually before the second decade with numbness and pain on the I--III digits. In most patients symptoms were unilateral but within 2 years they became bilateral. In all patients typical electrophysiological features of median nerve entrapment have been recorded. X-rays of the wrist showed narrow carpal tunnel in all patients. In all patients the possibility of having HNPP as well as familial amyloidosis has been excluded by molecular genetic and pathological studies. All patients underwent surgery and at postoperative stage symptoms were relieved or completely disappeared. Our study supports the theory that FCTS exists as a separate autonomic entity, therefore it is important in front of a sporadic case to investigate the family occurrence of CTS. PMID- 11903095 TI - Diffusion-weighted MR findings of central pontine and extrapontine myelinolysis. AB - Diffusion-weighted MR (DWI) can detect changes in water diffusion associated with cellular dysfunction, which enables the differentiation of cytotoxic edema from vasogenic edema. In this study on DWI findings in central pontine (CPM) and extrapontine myelinolysis (EPM), DWI showed high signal intensities in the bilateral pons, midbrain, and genu of the corpus callosum. The corresponding apparent diffusion coefficient values were rather low. This suggests that cytotoxic edema does in fact exist in CPM and EPM and that DWI can be useful in the rapid diagnosis and prediction of the various types of edema occurring in active demyelinating diseases. PMID- 11903094 TI - Blepharoclonus in multiple sclerosis. AB - OBJECTIVE: Keane described 2 patients with gaze-evoked blepharoclonus (BLC), a form of reflex BLC, and multiple sclerosis (MS). A search for common areas of demyelination and focal axonal atrophy (T1 black holes) of the central nervous system (CNS) in 11 patients with MS exhibiting eyelid closure BLC was conducted employing magnetic resonance imaging (MRI). Finding lesions in common CNS locations on these patients can help to elucidate the pathogenesis of this restricted movement disorder. MATERIALS AND METHODS: Eleven adult patients with relapsing-remitting, primary or secondary progressive MS were studied. MRI views were completed employing a 1.5-tesla scanner. Contrast Axial T1 imaging was obtained in 9 patients. RESULTS: TL blackholes were not identified. Ten patients had multiple, scattered periventricular (PV) areas of demyelination. Four patients exhibited brainstem lesions of diverse but inconsistent locations including midbrain, cerebellar peduncle, pons and medulla. In 2 of the patients the brainstem lesions were transient but BLC persisted after the lesions regressed. CONCLUSION: No common areas of CNS demyelination or focal axonal atrophy were identified on these patients with MS and BLC. The pathogenesis and clinical significance of BLC in MS remains to be elucidated. PMID- 11903096 TI - Tuberculous meningoencephalitis in HIV-seronegative patients: variety of clinical presentation and impact on diagnostics and treatment. AB - Tuberculous meningoencephalitis (TBM), an infrequent disease in Western European countries, shows a wide heterogeneity of clinical symptoms. MATERIAL AND METHODS: We present 4 patients (age range 42-72 years) with the definite diagnosis of isolated TBM. All patients were HIV-seronegative, only 1 patient was known to be immunoincompetent on admission due to acute myelocytic leukemia; other reasons for immune suppression were detected in 2 other patients (leukemia and idiopathic CD4+ T-lymphocytopenia, respectively). RESULTS: The diagnosis of TBM was confirmed in 3 cases by culture from CSF, in 1 case Mycobacterium tuberculosis was proven only in tracheal aspirate. In 1 patient M. bovis was found, which is an extremely rare cause of TBM in Germany. We report the contributions of different diagnostic tools (CSF analysis, neuroimaging) in reaching the presumptive diagnosis and in monitoring the further course. All patients developed neurological complications despite prompt tuberculostatic treatment. Three of the patients presented a chronic severe loss of consciousness of unclear origin. CONCLUSION: The possible causative relationships of these complications and their impact on the prognosis are discussed. PMID- 11903097 TI - Hypertrophic pachymeningitis with anti-neutrophil cytoplasmic antibody (p-ANCA), and diabetes insipidus. AB - We treated a patient with idiopathic cranial hypertrophic pachymeningitis and elevated serum titer of perinuclear anti-neutrophil cytoplasmic antibody (p-ANCA) reactive against myeloperoxidase. This 67-year-old man showed multiple cranial nerve-palsies, central diabetes insipidus (DI), and an intrasellar mass. DI and intrasellar mass had been present for 3 years, and DI had been well controlled by intranasal desmopressin. His nerve-palsies were most likely caused by thickened dura matter detected by the brain MRI. Granuloma may develop in the sella, and MRI findings in our patient are compatible to it. Corticosteroid and oral cyclophosphamide therapy improved his neurological symptoms and serum p-ANCA level with showing good correlation. DI improved temporally for 2 months. Few other cases of hypertrophic pachymeningitis with elevated p-ANCA have been reported, however the etiology is unknown. As p-ANCA antibodies have been detected in many of vasculitides, microvasculitis may be involved in some cases of idiopathic hypertrophic pachymeningitis. PMID- 11903098 TI - Machado-Joseph disease with retinal degeneration and dementia. AB - OBJECTIVES: To clarify the phenotypic varieties in Machado-Joseph disease (MJD). MATERIALS AND METHODS: We studied a 64-year-old man with ataxia, retinal degeneration and dementia neurologically, ophthalmologically and genetically. RESULTS: The patient noted dysesthesia of his hands at age 57 and later had memory disturbance. He had gait disturbance and needed a wheelchair at age 64. His total IQ was 61 on the WAIS-R. He had loss of central vision, ophthalmoplegia, hearing impairment, dysarthria, truncal and limb ataxia, sensory disturbance, and mild weakness of the extremities. Electrophysiologically he was suspected to have polyneuropathy. Brain MRI showed marked atrophy of the cerebellum and pons with mild cerebral atrophy. Ophthalmologic evaluation revealed multiple chorioretinal atrophy. Expanded CAG repeat numbers in MJD1 were 64. CONCLUSION: These findings indicate that the clinical features of MJD might cover a wider spectrum than previously expected, though it is possible that these complications, namely retinal degeneration and dementia, were incidental findings in this patient. PMID- 11903100 TI - Carotid Doppler - costs and need after stroke or TIA. AB - OBJECTIVES: To estimate the need for and the costs of carotid Doppler and carotid endarterectomy after stroke or TIA in non-selected hospitalized patients. MATERIAL AND METHODS: During 25 months hospitalized patients with stroke or TIA, in whom carotid endarterectomy could be relevant, were examined with carotid Doppler. If a significant stenosis was found, they were further evaluated for surgery. Based on our results, the requirement for future carotid endarterectomy and Doppler screening was estimated, and the costs of the procedures calculated. RESULTS: Among 1351 patients 703 were screened with carotid Doppler. Forty-five had severe (70-99%) stenosis of the relevant carotid artery. Only 3 were operated on. The future costs of screening were estimated under different assumptions. CONCLUSION: Carotid endarterectomy is expensive due to the large number of patients screened with carotid Doppler per operated patient. A careful clinical selection of patients for screening is necessary. PMID- 11903102 TI - Influence of benzodiazepines on antiparkinsonian drug treatment in levodopa users. AB - OBJECTIVES: Animal studies showed that benzodiazepines decrease the concentration of dopamine in the striatum. Benzodiazepines may therefore affect the treatment of Parkinson's disease. This study determined whether start of a benzodiazepine in patients on levodopa was followed by a faster increase of antiparkinsonian drug treatment. METHODS: Data came from the PHARMO database, which includes information on drug dispensing for all residents of six Dutch cities. Selected were all patients aged 55 years and older who used levodopa for at least 360 days. The rate of increase of antiparkinsonian drug treatment was compared between starters of a benzodiazepine and controls who did not start a benzodiazepine with the use of Cox's proportional hazard model. RESULTS: Identified were 45 benzodiazepine starters (27 women, mean age 76.4 years) and 169 controls (83 women, 74.3 years). Antiparkinsonian drug treatment increased faster in the benzodiazepine group; relative risk was 1.44 (95% confidence interval 0.80-2.59). CONCLUSION: This study has not found any statistically significant increase in antiparkinsonian drug treatment when a benzodiazepine was started in a small population of chronic levodopa users. PMID- 11903101 TI - Beta-CIT-SPECT combined with UPDRS appears to distinguish different parkinsonian conditions. AB - OBJECTIVES: An earlier study in l-dopa responding patients with idiopathic Parkinson's disease documented that progressive nigro-striatal degeneration shown with single photon emission computed tomography (SPECT) and the cocain analog iodine-123-beta-CIT (beta-CIT) correlated linearly with increasing motor scores in the Unified Parkinson's Disease Rating Scale (UPDRS). Here we have extended the study to include 2 tremor patients with mild parkinsonism, 2 poor l-dopa responders with parkisonism and 2 non l-dopa responders with severe parkinsonism. METHODS: SPECT scanning was performed 20 h after injection of beta-CIT and UPDRS was done at the time of beta-CIT injection. RESULTS: All patients in the present study showed less nigro-striatal degeneration in relation to UPDRS motor scores than the patients with idiopathic Parkinson's disease. CONCLUSION: The results suggest that the the combination of beta-CIT-SPECT and UPDRS motor scores has the potential to differentiate idiopathic Parkinson's disease from other parkinsonian conditions. PMID- 11903104 TI - Clinical utility of EEG in alcohol-related seizures. AB - OBJECTIVE: To study if electroencephalogram (EEG) can discriminate between alcohol-related seizures (ARS) and seizures unrelated to alcohol use. MATERIAL AND METHODS: Alcohol-related seizures was defined as a seizure in a patient with score > or = 8 in the Alcohol Use Disorders Identification Test (AUDIT). Twenty seven patients with ARS (22 without epilepsy: ARSwE), 21 AUDIT-negative epileptic patients with seizures (ES), and 30 other AUDIT negative patients with seizures (OS) were studied. Thirty-seven epilepsy outpatients and 79 sciatica inpatients were controls. RESULTS: Epileptiform and slow activity were less frequent in the ARSwE than in the ES group. Alpha amplitude was lower in the ARSwE than the other groups. Photoparoxysmal activity was not observed. EEG was associated with a larger negative predictive value (78% probability of non-ARS if EEG was abnormal) than a positive predictive value (55% probability of ARS if EEG was normal). CONCLUSION: A definitely abnormal EEG suggests epilepsy or symptomatic seizures unrelated to alcohol. The predictive value of a normal EEG is limited, but the typical post-ictal finding in ARS is nevertheless a normal low-amplitude EEG record. PMID- 11903103 TI - VNS therapy in clinical practice in children with refractory epilepsy. AB - OBJECTIVES: To study the efficacy, tolerability and safety of the vagus nerve stimulation (VNS) therapy in clinical practice, in 16 children and adolescents with refractory epilepsy. METHODOLOGY: We assessed the efficacy of VNS therapy, retrospectively by comparing seizure frequency, duration and severity at the time of most recent follow up (av: 24.9 months) to that in the 4 weeks prior to VNS surgery. Changes in quality of life, sleep and behaviour at last review was compared with that prior to VNS. Adverse effects elicited by specific questioning, spontaneous reporting and clinical examination are described. RESULTS: Vagus nerve stimulation resulted in a >50% reduction in seizure frequency in 62.5% of children with 25% achieving a >90% reduction. Vagus nerve stimulation was well tolerated in all but one of our cohort, with no serious side effects. CONCLUSION: Our results support its role as one of the options in intractable childhood epilepsy. PMID- 11903105 TI - The Vaga study of headache epidemiology II. Jabs: clinical manifestations. AB - OBJECTIVES: To describe clinical characteristics of cephalic jabs. MATERIAL AND METHODS: In a population study in Vaga, Norway, 1838 18-65-year-old-parishioners (88.6% of the target group) were subjected to a semi-structured interview, based on a headache questionnaire. RESULTS: Of 627 cases of jabs, 68% had single jabs; 4% volleys, and 28% a mixture of volleys and singlets. Most individuals had experienced only few jabs. Exceptionally, there were multiple attacks per day, even per hour. The ratio between an anterior and posterior location was 2.6. Neck movements and Valsalva manoeuvres seemingly occasionally precipitated attacks. Attacks were generally of mild/moderate intensity. Unilaterality prevailed over bilaterality; but unilateral pain might shift side. CONCLUSION: Cephalic jabs are generally solitary paroxysms, with rather long intervals between attacks. Jabs do not only occur in the trigeminal area. Occasionally vocalization and more frequently jolts accompany the paroxysm. PMID- 11903106 TI - Contribution of cerebellum and brainstem in the control of eye movement: evidence from a functional study in a clinical model. AB - The idiopathic cerebellar ataxias (IDCA) comprise a wide spectrum of neurodegenerative diseases with heterogeneous neuropathology, characterized by the negativity of search for any known genetic mutation. On the basis of both their clinical presentation and their magnetic resonance imaging pattern, patients with IDCA can be subdivided into patients with a purely cerebellar syndrome and atrophy of the cerebellum (IDCA-C) and patients with additional noncerebellar symptoms and atrophy of both cerebellum and brainstem (IDCA-P). The aim of the present study was to evaluate the disaggregated contribution of brainstem and cerebellum in the control of eye movements, by means of an extensive battery of quantitative tests covering most oculomotor subfunctions related to lesions of the cerebellum and the brainstem. The smooth-pursuit movement analysis showed a decrease in gain and magnitude in both subgroups of IDCA with respect to normal controls, without any significant differences in the prevalence pattern between the two subgroups; the mean values of these parameters, however, were significantly lower in IDCA-P than in IDCA-C subjects in both gain (P < 0.01) and magnitude (P < 0.001). No statistically significant difference was observed between the two subgroups in the analysis of saccadic movements or in the other parameters investigated. The distinction between IDCA-P and IDCA-C subgroups has clinical implications, as a poorer prognosis is related to brainstem involvement, which may occur late in the course of the disease. Thus, the possibility to detect the brainstem involvement, also in association with cerebellar impairment, by a relatively simple eye-movement analysis, potentially useful mainly in follow-up investigations, needs to be evaluated further. PMID- 11903107 TI - Dysphagia in multiple sclerosis - prevalence and prognostic factors. AB - The aim of the study was to analyse swallowing function and to identify reliable prognostic factors associated with dysphagia in a consecutive series of patients with multiple sclerosis (MS). Swallowing examination was performed by means of indirect and direct methods (fiberendoscopic evaluation) in 143 consecutive patients with primary and secondary progressive MS. Dysphagia was found in 49 patients (34.3%). A close relationship with dysphagia was found in the patients with severe brainstem impairment (OR=3.24; 95% CI 1.44-7.31) as compared to the patients without. There was also a significant correlation with pronounced severity of illness (OR=2.99; CI 1.36-6.59). Compensatory strategies were sufficient to resolve the dysphagia in 46 cases (93.8%). The potential risk of aspiration and malnutrition and the high efficacy of swallowing rehabilitation suggests that all MS patients should have a careful evaluation of deglutition functionality, especially those with brainstem impairment and a high grade of disability level. PMID- 11903108 TI - Autonomic function in demyelinating and axonal subtypes of Guillain-Barre syndrome. AB - OBJECTIVES: To investigate whether or not the pattern and extent of autonomic involvement differ between the two subtypes of Guillain-Barre syndrome (GBS), namely acute inflammatory demyelinating polyneuropathy (AIDP) and acute motor axonal neuropathy (AMAN). METHODS: Head-up tilt test, R-R interval variation, plasma noradrenaline concentration, skin vasomotor reflex (SVR) and sympathetic sweat response (SSwR) were used to estimate autonomic function in seven AIDP and eight AMAN patients. RESULTS: Heart rate and plasma noradrenaline concentration were significantly high in the AIDP group but not in the AMAN group. Skin vasomotor reflexes were generally preserved and SSwRs were impaired in patients with severe neurological deficits for both AIDP and AMAN groups. CONCLUSION: The patterns of autonomic involvement are qualitatively different between AIDP and AMAN. Acute inflammatory demyelinating polyneuropathy is characterized by cardio sympathetic hyperactivity, excessive or reduced sudomotor function and preserved skin vasomotor function, while AMAN is not necessarily generally associated with marked autonomic dysfunction except for the sudomotor hypofunction seen in patients with severe neurological deficits. PMID- 11903109 TI - Recognition memory for emotional pictures in Alzheimer's patients. AB - OBJECTIVE: The purpose of the present study was to examine whether Alzheimer's Disease (AD) patients can benefit from the emotional content of visual stimuli in a picture recognition test. METHODS: Sixteen patients with AD and 19 normal controls matched for age and years of education, were studied. Sixteen pictures (with varying emotional contents) were presented to each participant. Thirty minutes later, a recognition test was applied with the target-pictures mixed among 34 others of similar content. The subjects were instructed to rate them as pleasant, unpleasant or indifferent. RESULTS: The total of pictures correctly recognized by the AD patients (75.4% of the target-pictures) was smaller than that of the controls (96.4%). Controls recognized more emotional pictures than indifferent pictures. CONCLUSION: Emotional content enhanced recognition of pictures in normal subjects, whereas for the Alzheimer's subjects the emotional significance attached to the pictures was of no benefit to enhance recognition. PMID- 11903110 TI - CTG repeat polymorphism in DMPK gene in healthy Yugoslav population. AB - OBJECTIVES: Myotonic dystrophy type 1 (DM1) is caused by large expansions of cytosine-thymine-guanine (CTG)-repeats in myotonic dystrophy protein kinase (DMPK)-gene. This gene is highly polymorphic in healthy individuals. It has been proposed that expanded alleles originated from the group of large sized normal alleles. If this is correct, one should expect a positive correlation between the frequency of large sized normal alleles and a prevalence of this disorder in a population. In this paper we determined the distribution of alleles of DMPK gene in healthy Yugoslav population. MATERIAL AND METHODS: A sample of 235 healthy individuals of Yugoslav origin have been genotyped for the alleles of DMPK locus. RESULTS: We found 22 different alleles, ranging in size from 5 to 29 repeats. Among 470 chromosomes studied, 41 chromosomes had more than 18 repeats (8.72%). CONCLUSIONS: Relatively high frequency of large sized normal alleles found in our population, suggest that prevalence of DM1 in Yugoslavia should not be different from the prevalence in other European populations. PMID- 11903111 TI - Cerebrovascular reactivity in patients under long-term acetazolamide treatment. AB - BACKGROUND AND PURPOSE: The acetazolamide (AZA) test is a well-accepted method for measuring the vascular reactivity of the cerebral arteries. In order to investigate the nature of this reactivity after long-term daily AZA treatment, the cerebral blood velocity (CBV) was measured using transcranial Doppler in patients under continuous AZA treatment after a single AZA 1 g intravenous (IV) dose. METHODS: Thirteen patients (eight women, five men) on long-term daily AZA (750 mg/day, mean treatment duration 68 +/- 12+ months) were included in the study. The CBV of the middle cerebral artery (MCA) and the basilar artery (BA), including the values of peak velocity, mean velocity and pulsatility index (PI) were measured. The examination was performed twice - with the initial IV administration of AZA and 20 min later. The results were compared with those of 10 age matched volunteers. RESULTS: A consistent significant increase of CBV in the right and left MCA (P < 0.001 for both arteries) was found in all study participants. A highly significant decrease of peak CBV in the BA (P < 0.001) was found in the post-AZA velocities of the patient's group. In the control group, a consistent significant increase in all post-AZA tests was demonstrated (P < 0.001). CONCLUSIONS: A mild elevation of blood velocity in the MCAs concomitant with a highly significant decrease of velocity in the BA was present in all examined patients. These patterns of CBV changes indicate the presence of a 'steal phenomenon' from the posterior to the anterior circulation and stress the necessity for caution when evaluating the indications for performance of the AZA test in patients under continuous AZA therapy. PMID- 11903113 TI - Long term follow up of a hemimasticatory spasm. AB - OBJECTIVES: To describe the clinical and neurophysiological findings in a case of hemimasticatory spasm (HMS) followed during 14 years of evolution. MATERIAL AND METHODS: A woman suffered from very frequent paroxysmal episodes of painful involuntary occlusion of the jaw. Neurophysiological studies were performed at the 3, 12 and 14 years after the onset of symptoms. They included a needle electromyographic (EMG) evaluation of the main jaw closing and opening muscles, the jaw reflex (JR), the masseteric silent period (MSP) and the masseteric inhibitory reflex (MIR). RESULTS: Clinical symptoms remained unchanged throughout the period of observation. Conventional EMG never disclosed neurogenic signs. Voluntary closure of the jaw systematically provoked an abnormal activity with muscle cramps characteristics, restricted to the left masseter muscle. Left JR response was normal in the first evaluation and became delayed and of reduced amplitude in the second. The MSP and MIR were abolished on the left side during the spasmodic episodes whereas they were strictly normal out of them. The MIR abnormalities showed the characteristic pattern of an efferent lesional type. CONCLUSIONS: Hemimasticatory spasm probably is the consequence of an abnormal trigeminal hyperexcitability likely induced by the demyelinating lesion of its peripheral motor pathway. The main neurophysiological abnormalities may persist unmodified over a long course of the disease and allow the differential diagnosis of HMS from oromandibular dystonia and temporomandibular dysfunction (TMD). PMID- 11903112 TI - Leukotrienes in patients with clinically active multiple sclerosis. AB - OBJECTIVES: The role of leukotrienes (LTs) in the pathophysiology of multiple sclerosis (MS) has been controversially discussed in the past. Studies of LTs in the cerebrospinal fluid (CSF) revealed different results mainly because of analytical difficulties. MATERIAL AND METHODS: In the present study we used highly sensitive and specific analytical methods for measuring LTs in the CSF as well as in urine samples from 20 patients with active MS and 20 control patients with noninflammatory neurological disorders. RESULTS: LTB4 concentrations in CSF were almost twice as high in MS patients compared with controls (P < 0.001). CSF concentrations of the cysteinyl-LTs (LTC4, LTD4 and LTE4) as well as urinary LTE4 showed no significant differences compared with controls (P > 0.05). In addition, there was no significant association between CSF pleocytosis, clinical severity or time of disease onset. CONCLUSIONS: The increased concentration of LTB4 in the CSF of MS patients may indicate a biological importance for this mediator in MS. PMID- 11903115 TI - Familial and environmental risk factors in Parkinson's disease: a case-control study in north-east Italy. AB - BACKGROUND AND OBJECTIVE: The aetiology of Parkinson's disease remains unknown, although both genetic susceptibility and environmental factors are considered putative contributors to its origin. We performed a case-control study to investigate the association of familial and environmental risk factors with Parkinson's disease (PD). METHODS: We studied 136 patients with neurologist confirmed PD and 272 age- and sex-matched controls, affected by neurological diseases not related to PD. The risk of developing idiopathic PD associated with the following familial and environmental factors: positive family history of PD, positive family history of essential tremor (ET), age of mother at subject's birth, rural birth, rural living, well water use, farming as an occupation, exposure to pesticides, head tremor, exposure to general anaesthesia and to ionizing radiations, food restriction, concentration camp imprisonment and smoking has been assessed by using univariate and multivariate statistical techniques. RESULTS: In the conditional multiple logistic regression analysis, positive family history of PD (OR 41.7, 95% CI 12.2-142.5, P < 0.0001), positive family history of ET (OR 10.8, 95% CI 2.6-43.7, P < 0.0001), age of mother at subject's birth (OR 2.6, 95% CI 1.4-3.7, P=0.0013), exposure to general anaesthesia (OR 2.2, 95% CI 1.3-3.8, P=0.0024), farming as an occupation (OR 7.7, 95% CI 1.4-44.1, P=0.0212) and well water use (OR 2.0, 95% CI 1.1-3.6, P=0.0308) exhibited a significant positive association with PD, whereas smoking showed a trend toward an inverse relationship with PD (OR 0.7, 95% CI 0.4-1.1, P < 0.06). CONCLUSIONS: We conclude that both familial and environmental factors may contribute to PD aetiology. PMID- 11903116 TI - Long-term intrathecal Baclofen infusion in supraspinal spasticity of adulthood. AB - OBJECTIVE: To evaluate long-term results of chronic intrathecal Baclofen infusion on the spasticity, on the spasms and to evaluate the side-effects of the intrathecal Baclofen in patients with supraspinal spasticity. CLINICAL MATERIALS AND METHODS: Fourteen patients with severe progressive refractory to medical therapy spasticity were evaluated after chronic intrathecal Baclofen infusion performed by implantation of subcutaneous pump. The patients had suffered traumatic or anoxic acquired brain injuries. The clinical evaluation was made using Ashworth Scale (AS) and the Spasm Frequency Scale (SFS). RESULTS: The intrathecal therapy showed a statistically significant improvement of the tone and of the spasms. CONCLUSIONS: The intrathecal infusion of Baclofen seems to be an effective treatment in patients with supraspinal spasticity. PMID- 11903117 TI - Varicella, ephemeral breastfeeding and eczema as risk factors for multiple sclerosis in Mexicans. AB - OBJECTIVE: It has been suggested that the incidence of multiple sclerosis (MS) in Mexico and other countries of Latin America has increased steadily for the last two decades. We made a thorough search of antecedents on MS patients that could be potential risk factors. METHODS: A case-control study was conducted using a questionnaire that included demographic, nutritional, infectious and personal antecedents previously identified in other reports as possible risk factors for MS. RESULTS: The frequency of varicella, ephemeral breastfeeding and eczema in the medical history of MS patients were significant when compared with controls; all appeared to be mutually additive. However, they were unrelated with clinical characteristics or disease severity. CONCLUSION: During the last decades, breastfeeding has been abandoned in large segments of society and the incidence of varicella and childhood eczema keeps a north-south gradient similar to that described for MS. These factors may participate in the sharp increase of MS in countries like Mexico traditionally considered as an area of very low incidence. PMID- 11903119 TI - Prognostic factors in stroke rehabilitation: the possible role of pharmacological treatment. AB - OBJECTIVES: The aim of the present study was to determine the impact of commonly used and potentially detrimental drugs on rehabilitation results and to clarify their role as prognostic factors. MATERIAL AND METHODS: The study included 154 patients admitted to a rehabilitation hospital for sequelae of a first stroke. Multivariate analyses were performed using effectiveness of treatment, evaluated by both the Barthel Index (BI) and the Rivermead Mobility Index (RMI) and low response on both of these indexes as dependent variables. Independent variables were medical, demographic and pharmacological factors. RESULTS: The use of detrimental drugs was negatively associated with effectiveness on both BI and RMI. Severity of stroke (Canadian Neurological Scale score at admission) and hemineglect were the other negative prognostic factors that significantly entered the analyses. On the other hand, the presence of Broca's aphasia positively influenced the recovery, essentially due to prolonged length of stay. The presence of detrimental drugs and hemineglect were associated with a higher risk of low response on both BI and RMI. CONCLUSION: These findings confirm that the use of some drugs can influence rehabilitation results. Therefore, the choice of pharmacological treatment of stroke patients should be carefully evaluated by considering the potential detrimental effects of some drugs commonly used for the treatment of coincidental medical conditions. PMID- 11903118 TI - Prevalence of herpesvirus DNA in MS patients and healthy blood donors. AB - OBJECTIVES: The aim of this study is to determine the DNA prevalence of different members of Herpesviridae in multiple sclerosis (MS) patients and to describe the possible effect of beta-interferon treatment on such prevalence. MATERIAL AND METHODS: With a nested polymerase chain reaction (PCR) assay we have studied the DNA of the peripheral blood mononuclear cells (PBMCs) of 204 whole blood samples, [102 from patients with relapsing-remitting MS (RRMS), of which 62 were treated with beta-interferon, and 102 from healthy blood donors]. RESULTS: We only found a statistically significant difference for human herpesvirus type 6 (HHV-6) DNA prevalence (P < 0.0001): HHV-6 is 2.26 times more frequent in MS patients than in healthy donors. There was no difference in the HHV-6 prevalence between beta interferon treated and untreated patients. CONCLUSION: 1. Among the herpesviruses, HHV-6 was the only one showing altered prevalence. This either indicates that HHV-6 is involved in the pathogenesis of MS, or it simply indicates that MS influences latency or reactivation of HHV-6 without any direct involvement of HHV-6 in the disease process of MS. 2. Treatment with beta interferon does not make a difference on the DNA prevalence of the herpesviruses studied in our MS patients. PMID- 11903120 TI - Natural anticoagulants (antithrombin III, protein C, and protein S) in patients with mild to moderate ischemic stroke. AB - BACKGROUND AND PURPOSE: The role of the natural anticoagulants, antithrombin III (AT III), protein C (PC), and protein S (PS), in patients with mild to moderate ischemic stroke remains uncertain. We aimed to find out whether their levels in peripheral blood correlated with the severity of neurological deficit or can predict clinical outcome and recurrence. METHODS: We studied AT III, PC, and free PS levels in 55 consecutive patients likely to survive the study period on admission, 1 week, 1 month and 3 months after a first-ever ischemic stroke. Sex- and age-matched controls were studied once. All patients underwent a full neurological examination and blood sampling at each study time point; comprehensive stroke risk factors were recorded, and the etiology of the ischemic stroke was determined. All patients were contacted 3 years later for possible recurrent ischemic events. RESULTS: AT III level was found to be significantly lower at all time points after stroke; PC level was significantly increased on admission and normal at subsequent measurements, and PS level was normal on admission but significantly decreased later. The levels of the natural anticoagulants did not correlate with the etiology of stroke, any stroke risk factor, or neurological scores, except that the AT III level on admission showed significant correlation with stroke severity and disability at 3 months. Natural anticoagulant levels did not predict recurrence of ischemic stroke. CONCLUSIONS: The measurements of the level of AT III, PC, or PS did not deliver useful information for management of patients with mild or moderate ischemic stroke, expect that AT III level on admission might predict outcome. PMID- 11903121 TI - Primary intraventricular haemorrhage in adults. AB - INTRODUCTION: Primary intraventricular haemorrhage (PIVH) is an uncommon type of intracerebral haemorrhage. Relatively little is known about clinical and imaging features, and even less about prognosis and predictors of mortality. MATERIAL AND METHODS: We analysed clinical and imaging features, causative factors and outcome of 26 patients with CT brain scan evidence of PIVH. A multivariate regression model of failure time data was used to assess predictors of in-hospital mortality. RESULTS: Loss of consciousness was the first manifestation of PIVH in six patients and occurred after all other symptoms in five. In other patients, onset was characterized by headache, vomiting, confusion and disorientation (n=8) or by headache with or without vomiting (n=7). Angiography revealed vascular malformations in eight patients (31%). Other possible causative factors were clotting disorder in one patient and arterial hypertension in 10. No cause was identified in seven patients. Early hydrocephalus was the most frequent complication and resolved spontaneously in a minority of patients. In-hospital mortality was high (42%): four patients died early of direct consequence of bleeding and seven died after clinical worsening because of increasing hydrocephalus or other adverse events. Multivariate analysis indicated Glasgow Coma Scale < or = 8 (OR 4.67; 95% CI 1.22-17.92) and early hydrocephalus (OR 4.93; 95% CI 1.13-21.59) as independent predictors of in-hospital mortality. CONCLUSION: In patients with PIVH, hydrocephalus seems to be a critical determinant of in-hospital mortality and this suggests the need for early treatment strategies. PMID- 11903122 TI - Drug-associated headache is unrecognized in patients treated at a neurological centre. AB - OBJECTIVE: We studied the frequency of unrecognized headache associated with overuse of analgesic drugs in a population of headache patients treated at a neurological centre. METHODS: Patients in North Norway referred to a neurologist for headache during a 2-year period completed a questionnaire. From a total of 945, 262 patients (28%) reported headache 3 days or more per week and used analgesic drugs on a daily bases. RESULTS: A specific diagnoses given by the neurologist was reported in 134 of the patients (51%). Only two patients reported that they suffered from a possible drug-associated headache. CONCLUSION: This study shows that drug overuse may be the cause of chronic headache in more than 1/4 patients referred to neurologists. Drug-associated headache is a difficult diagnosis which deserves more attention because it is a common and treatable condition. PMID- 11903123 TI - Is squatting a triggering factor for stroke in Indians? AB - BACKGROUND: Undocumented observations of ours suggested increased occurrence of strokes in the early hours of the morning and in the toilets where most Indians still squat. The present study has been designed to assess the validity of this observation and to identify the likely triggering factor in stroke onset in such situations. MATERIAL AND METHODS: Part A of the study looked into stroke onset data of 100 consecutive CT-confirmed stroke patients specially in relation to time of day, place, posture and activity of the individual. Part B of the study included 67 healthy volunteers (age 24-49 years) whose supine and squatting blood pressure (BP) were measured and the differences noted. Part C of the study consisted of repeating the same procedure in 104 known hypertensives on treatment (age 28-60 years). RESULTS: Part A of the study revealed that most strokes (52%) occurred in the morning and at home (86%) and over a third (36%) while in toilets. Thirty-six per cent of the strokes occurred while the subjects squatted, mostly during the act of defecation. More than half of the haemorrhagic strokes occurred while the subjects were in squatting position. In normal healthy volunteers squatting produced a small (8.09 +/- 7.04 mmHg) but significant rise in systolic blood pressure (SBP) but not in diastolic blood pressure (DBP) (0.9 +/- 7.38 mmHg). In contrast, in treated hypertensives squatting produced a significantly higher rise in both SBP (14.49 +/- 11.63 mmHg) and DBP(9.10 +/- 9.19 mmHg). CONCLUSION: The relationship of the clinical observations regarding stroke onset with the BP changes noted on squatting in healthy as well as hypertensive subjects appears to be more than fortuitous. Squatting induced rise in BP appears to be an important triggering factor for stroke onset in subjects at risk in India. PMID- 11903125 TI - Diffusion-weighted magnetic resonance imaging in Wernicke's encephalopathy. AB - OBJECTIVE: To report diffusion-weighted imaging (DWI) findings and postulate the pathogenic mechanism of Wernicke's encephalopathy (WE). PATIENT: A 47-year-old woman presented with altered consciousness, ophthalmoplegia, and ataxia. DWI revealed the abnormal signal changes in periaqueductal gray matter, mamillary bodies and bilateral medial thalami. Apparent diffusion coefficient (ADC) map revealed the high signal intensity lesions in bilateral medial thalami, suggestive of vasogenic edema. The abnormal signal intensity lesions disappeared on follow-up imaging with clinical improvement. CONCLUSIONS: Vasogenic edema plays an important role in the pathogenesis of WE and can be reversed by proper management. DWI findings in the early stage of WE may provide useful information about the prognosis. PMID- 11903124 TI - Reversible peripheral neuropathy in idiopathic hypoparathyroidism. AB - We describe a 40-year-old male with idiopathic hypoparathyroidism presenting with tetany, proximal weakness, signs of hypocalcaemia including Chvostek and Trousseau's and diminished tendon reflexes in the upper and lower limbs. Electrophysiological studies revealed a sensory-motor neuropathy, predominantly axonal as evidenced by decreased CMAP amplitudes, with normal distal latencies velocites, except for median nerve where a prolonged distal latency was observed. Serial nerve conduction studies were performed at repeated intervals for 2 years, while he received treatment for hypoparathyroidism (calcium and vitamin D supplementation). A progressive improvement in neuropathy both clinical and on electrophysiological studies was observed. Occurrence of peripheral neuropathy in hypocalcaemic states such as hypoparathyroidism and its reversibility after normalization of calcium homeostasis lend proof to the role of critical Ca2+ ion concentration in the normal functioning of the peripheral axons. PMID- 11903126 TI - Effects of long-term inhibition of neuronal nitric oxide synthase on blood pressure and renin release. AB - Nitric oxide (NO) produced by neuronal NO-synthase (nNOS) in macula densa cells may be involved in the control of renin release. 7-Nitro indazole (7-NI) inhibits nNOS, and we investigated the effect of short- (4 days) and long-term (4 weeks) 7 NI treatment on blood pressure (BP), plasma renin concentration (PRC) and glomerular filtration rate (GFR) in rats on different salt diets. Rats were divided into three groups and given low-salt (LS), normal (C) and high-salt (HS) diets. Each diet group was subdivided into two groups treated either with 7-NI or vehicle. Long-term 7-NI-treated rats (LS and C) showed increased BP compared with controls (LS: 149 +/- 4 vs. 133 +/- 3; C: 146 +/- 4 vs. 127 +/- 4 mmHg). Blood pressure in HS rats did not differ from that in controls. Plasma renin concentration was stimulated in LS-rats (251 +/- 64 mGU mL(-1)) compared with C and HS rats (42 +/- 8 and 39 +/- 5 mGU mL(-1), respectively) but was not significantly affected by chronic 7-NI treatment (350 +/- 103, 49 +/- 10 and 50 +/- 15 mGU mL(-1) in LS, C and HS, respectively). In rats treated with 7-NI for 4 days, no effect on BP was seen, but PRC was increased in 7-NI treated LS rats compared with vehicle treated LS rats (107 +/- 15 vs. 56 +/- 1 mGU mL(-1)). Stimulation of PRC in LS rats was further enhanced by 7-NI after 4 days of treatment, but not affected in rats treated for 4 weeks. This suggests that inhibition of nNOS stimulates renin release but that this stimulatory effect in the long run might be depressed by the increase in blood pressure. PMID- 11903127 TI - Modifications of microvascular filtration capacity in human limbs by training and electrical stimulation. AB - This study investigated whether an increase in microvascular surface area as a result of endurance training, which increases human skeletal muscle capillarity, would translate to greater capacity for fluid filtration compared with strength training, which does not affect capillary supply. Values for filtration capacity, Kf, derived from the slope of calf volume change, Jv, measured by venous occlusion plethysmography, against cuff pressure during a protocol of small cumulative pressure steps, were significantly higher in endurance athletes (5.78 +/- 0.88 mL min(-1) 100 mL(-1) mmHg(-1) x 10(-3), P < 0.05) than controls (3.38 +/- 0.32 mL min(-1) 100 mL(-1) mmHg(-1) x 10(-3) whereas strength-trained athletes had values similar to control (4.08 +/- 0.56 mL min(-1) 100 mL(-1) mmHg( 1) x 10(-3), ns), suggesting that surface area is important. However, when sedentary subjects underwent either a 4-week unilateral dynamic plantarflexion training programme (70% peak power, 20 min day(-1), 5 days week(-1) or a calf muscle electrical stimulation programme (8 Hz, 3 x 20 min day(-1), 5 days week( 1), neither of which caused limb blood flow to alter after training nor would be expected to increase capillarity, only the stimulation group showed a significant increase in Kf (6.68 +/- 0.62 mL min(-1) 100 mL(-1) mmHg(-1) x 10(-3) post training vs. 3.38 +/- 0.38 pre-training, P < 0.05). This may be because stimulation enhances perfusion preferentially to glycolytic fibres, or maintains high levels of vascular endothelial growth factor (VEGF) or changes lymph clearance. PMID- 11903128 TI - Concentric force enhancement during human movement. AB - In order to understand the possible mechanisms contributing to enhanced concentric performance in stretch-shortening cycle exercises in vivo the present study examined knee extension torque, electromyogram (EMG) activity and fascicle length of the vastus lateralis muscle in maximal and submaximal human movements. Maximal concentric knee extensions (120 degrees s(-1)) were done after pre stretch and pre-isometric conditions by nine volunteers. During shortening at the knee angle of 115 degrees (180 degrees = extended) the knee extension torque was found to be greater in pre-stretch condition (272 vs. 248 N m, P < 0.05) although the torque level prior to shortening was smaller than in pre-isometric condition (268 vs. 314 N m, P < 0.05). At the moment of torque enhancement the EMG activity levels or fascicle lengths did not differ between the conditions. It is proposed that besides specific experimental conditions the present enhancement may be related to longer fascicle length prior to shortening (by 4.1 cm, P < 0.05) in pre-stretch condition and to modified length-tension properties. Fascicle length behaviour was found to play an important role also in unilateral, submaximal sledge-jump conditions where pre-loading was altered but the concentric net impulse and joint angular movements were the same. In repeated drop jumps with greater pre-load the changes in fascicle length were smaller than in the counter movement jump that was characterized by a lower force and activity level in the eccentric phase. Results from the present maximal and submaximal loading conditions suggest that the benefits of stretch-shortening cycle muscle function may come through different interactive mechanisms that may be task specific. PMID- 11903129 TI - Insulin action on rates of muscle protein synthesis following eccentric, muscle damaging contractions. AB - The purpose of this study was to determine whether eccentric, muscle-damaging contractions affect insulin action on muscle protein synthesis. Male Wistar rats (n = 28) were anaesthetized either once or twice separated by 7 days' rest, and one limb was electrically stimulated to contract eccentrically, while the contralateral limb served as a non-stimulated control. Twenty-four and 48 h after contractions, rates of protein synthesis were assessed in soleus and red or white gastrocnemius muscles during a hindlimb perfusion with or without insulin (20 000 microU mL(-1)). Rates of protein synthesis were not different in non-stimulated muscle, with or without insulin (P > 0.05). In red or white gastrocnemius without insulin, rates of protein synthesis were significantly reduced (P < 0.05) 24 and 48 h after a single session and 48 h after a double session of muscle contractions. However, protein synthesis was normalized with insulin 24 and 48 h after contractions in red, and 48 h after contractions in white gastrocnemius. In soleus muscle, protein synthesis was impaired only 48 h after the second session, but partially restored by insulin (P < 0.05). These results indicate that muscle becomes more sensitive to insulin action on rates of protein synthesis after muscle-damaging contractions. PMID- 11903130 TI - Effect of glycogen loading on skeletal muscle cross-sectional area and T2 relaxation time. AB - This study was performed to investigate if glycogen loading of skeletal muscles, by binding water, would effect the cross-sectional area (CSA) and if an altered water content would alter the transverse relaxation time (T2) measured by magnetic resonance imaging (MRI). Five healthy volunteers participated in a programme with 4 days of extremely carbohydrate-restricted meals followed by 4 days of extremely high carbohydrate intake. The CSA and T2 of thigh and calf muscles were related to the intramuscular glycogen content evaluated at days 4 and 8. An increase in glycogen content from 281 to 634 mmol kg(-1) dry wt increased the CSA of the vastus muscles by 3.5% from 78 +/- 11 to 80 +/- 12 cm2 and the thigh circumference by 2.5% from 146 +/- 20 to 150 23 cm2. Calf circumference increased non-significantly by 4% from 78 +/- 15 to 82 +/- 19 cm2. Mono-exponential T2 decreased in m. tibialis anterior from 27.8 +/- 1.2 to 26.9 +/- 1.7 ms, did not change in m. vastus lateralis 26.5 +/- 1.9 ms/26.6 +/- 1.3 ms or in m. gastrocnemius 29.5 +/- 1.0 ms/29.8 +/- 1.9 ms. Glycogen loading increased the signal intensity mainly at different echo times (TE) 15 and 30 ms. The study shows that increased glycogen filling in the muscles increases muscle CSA and that this can be detected by MRI. The signal intensity increased the most at shorter TEs suggesting a more tight intracellular binding of water in glycogen loaded muscles. PMID- 11903131 TI - Cyclopiazonic acid and thapsigargin reduce Ca2+ influx in frog skeletal muscle fibres as a result of Ca2+ store depletion. AB - We have investigated the influence of the sarcoplasmic reticulum (SR) Ca2+ content on the retrograde control of skeletal muscle L-type Ca2+ channels activity by ryanodine receptors (RyR). The effects of cyclopiazonic acid (CPA) and thapsigargin (TG), two structurally unrelated inhibitors of SR Ca(2+) adenosine triphosphatase (ATPase), were examined on the SR Ca2+ content, the calcium current and contraction in single frog semitendinosus fibres using the double mannitol-gap technique. At moderate concentrations that only partially inhibited Ca2+ sequestration by the SR, CPA (2-4 microM) induces a concentration dependent depression of contraction and Ca2+ current amplitudes. When Ba2+ is the charge carrier, the inward current is not changed by CPA suggesting that this Ca(2+)-pump inhibitor does not directly affect dihydropyridine Ca2+ channels. Similar effects were obtained with TG (1-5 microM). Changes in Ca2+ currents and contraction were accompanied by a reduced Ca2+ loading of the SR. We attribute the modulation of the Ca2+ current to the selective inhibition of the SR Ca2+ ATPase, resulting in a decreased Ca2+ release and thereby a reduced activation of calcium inward currents. This is therefore taken to represent a calcium release dependent modulation of skeletal muscle L-type Ca2+ channels. PMID- 11903132 TI - Inhibition of human alpha1E subunit-mediated ca2+ channels by the antipsychotic agent chlorpromazine. AB - Chlorpromazine is a neuroleptic antipsychotic agent with a long history of clinical use. Its primary mode of action is thought to be through modulation of monoaminergic inter-neuronal communication; however, its side-effect profile indicates substantial activities in other systems. Recent work has begun to uncover actions of this compound on ion channels. In this light we have investigated the actions of chlorpromazine on the recombinant alpha1E subunit encoded voltage-sensitive Ca2+ channel (VSCC) that is believed to encode drug resistant R-type currents found in neurones and other cells. Chlorpromazine produced a dose-dependent antagonism of these channels that was reversed on drug removal. The mean IC50 was close to 10 microM. At this concentration, the level of antagonism observed was dependent on the membrane potential, with greater inhibition being observed at more negative test potentials. Furthermore, chlorpromazine induced substantial changes in the steady-state inactivation properties of alpha1Ebeta3-mediated currents, although it was not seen to elicit a corresponding change in inactivation kinetics. These results are discussed with regard to the possible clinical mechanisms of chlorpromazine actions. PMID- 11903133 TI - Reduced autonomic activity during stepwise exposure to high altitude. AB - Several studies have shown increased sympathetic activity during acute exposure to hypobaric hypoxia. In a recent field study we found reduced plasma catecholamines during the first days after a stepwise ascent to high altitude. In the present study 14 subjects were exposed to a simulated ascent in a hypobaric chamber to test the hypothesis of a temporary reduction in autonomic activity. The altitude was increased stepwise to 4500 m over 3 days. Heart rate variability (HRV) was assessed continuously in seven subjects. Baroreceptor reflex sensitivity (BRS) was determined in eight subjects with the 'Transfer Function' method at baseline, at 4500 m and after returning to baseline. Resting plasma catecholamines and cardiovascular- and plasma catecholamine- responses to cold pressor- (CPT) and mental stress-test (MST) were assessed daily in all and 12 subjects, respectively. Data are mean +/- SEM. Compared with baseline at 4500 m there were lower total power (TP) (35 457 +/- 26 302 vs. 15 001 +/- 11 176 ms2), low frequency (LF) power (3112 +/- 809 vs. 1741 +/- 604 ms2), high frequency (HF) power (1466 +/- 520 vs. 459 +/- 189 ms2) and HF normalized units (46 +/- 0.007 vs. 44 +/- 0.006%), P < or = 0.001. Baroreceptor reflex sensitivity decreased (15.6 +/- 2.1 vs. 9.5 +/- 2.6 ms mmHg(-1), P = 0.015). Resting noradrenaline (NA) decreased (522 +/- 98 vs. 357 +/- 60 pmol L(-1), P = 0.027). The increase in systolic blood pressure (SBP) and NA during mental stress was less pronounced (21 +/- 4 vs. 10 +/- 2% and 25 +/- 9 vs. -2 +/- 8%, respectively, P < 0.05). The increase in SBP during cold pressor test decreased (16 +/- 3 vs. 1 +/- 6%, P = 0.03). Diastolic blood pressure, HR and adrenaline displayed similar tendencies. We conclude that a transient reduction in parasympathetic and sympathetic activity was demonstrated during stepwise exposure to high altitude. PMID- 11903135 TI - Dynamic cardiomyoplasty: a new summing up. PMID- 11903136 TI - Altruism versus reality: an academic medical center's role in device development. PMID- 11903138 TI - Risk of bacterial infection in patients under intravenous iron therapy: dose versus length of treatment. AB - Some studies have suggested that intravenous iron therapy may be associated with an increased risk of infection. We analyzed the incidence of bacterial infection in 111 hemodialysis patients. Group 1 (n = 39, transferrin saturation <20%) received 10 doses of 100 mg of intravenous iron saccharate, 3 doses per week (28 treatment days); Group 2 (n = 13, transferrin saturation <20%) received 20 doses, 3 doses per week (70 treatment days); and Group 3 (n = 59, transferrin saturation 20-50%) received 10 doses, 1 dose per week (70 treatment days). The follow-up was 150 days for all groups, and all infectious episodes were recorded. Pulmonary infection was the most frequent event observed in all of the groups. In an incidence-density analysis, Group 2, which received a total of 20 doses, presented a significantly higher incidence of infection than Group 3, which received only 10 doses over the same period (0.13 versus 0.06 infections per patient per month, p = 0.04). No difference was observed between Groups 1 and 2 suggesting that the risk of infection during iron therapy is dose dependent rather than time length dependent. PMID- 11903139 TI - Customized bicarbonate buffered dialysate and replacement solutions for continuous renal replacement therapies: effect of crystallization on the measured levels of electrolytes and buffer. AB - The aim of this study was to investigate the impact of the addition of calcium to bicarbonate solutions for continuous renal replacement therapy (CRRT). We tested single bag (bicarbonate and calcium mixed 24 h before testing) and double bag solutions (mixed immediately before) with and without the addition of 4 mEq/L of acetate. Prescribed calcium varied from 0 to 5 mEq/L. All test solutions containing calcium showed crystallization at light microscopy. The double bag solutions decreased but did not prevent crystallization. The addition of acetate did not interfere with crystallization. Crystallization, as measured by the weight of the crystals after filtration of the solutions, showed a significant positive correlation with the calcium deficit (prescribed minus measured) and with partial pressure of carbon dioxide. The measured level of calcium was lower than expected and correlated with crystallization. Our results suggest that the use of bicarbonate solutions containing calcium as replacement fluids for CRRT is a potentially unsafe procedure. PMID- 11903141 TI - Different low constant flows can equally determine the lower inflection point in acute respiratory distress syndrome patients. AB - Among the possible techniques to obtain the pressure-volume (P x V) curve at the bedside the low constant flow (CF) is the easiest and quickest one. However, the best value for CF to perform a good semi-static P x V curve is still to be determined. The purpose of this study was to evaluate the influence of 4 different CFs (1, 2, 5, and 10 L/min) on determination of lower inflection point of the P x V curve (L-Pflex) and upper inflection point of the P x V curve (U Pflex) on the maximum slope and on the inspiratory work of breathing (up to volume of 1.35 L; inspiratory work L/cm H2O), comparing the volume estimated from the CF with the measured volume obtained by the respiratory inductive plethysmograph. The design was a prospective study, and the setting was an adult medical intensive care unit of a university hospital. There were 7 acute respiratory distress syndrome (ARDS) patients, less than 5 days of installation, after the standardization of lung volume history received sequentially from 4 different low inspiratory CFs in 2 trials. The P x V curve lasted from 73 +/- 1.6 s (1 L/min) to 8.8 +/- 0.69 s (10 L/min). The L-Pflex differed in the 2 performed trials (p = 0.04). There was no difference of L-Pflex among the 4 CFs comparing the 3 methods (p = 0.072) used for its calculation as well as comparing the estimated and the measured volume (p = 0.456). The maximum slope decreased significantly while increasing the flow from 1 to 10 L/min just in the estimated volume (p = 0.03). The inspiratory work did not increase with the increment of the flow either in the estimated volume (p = 0.217) or in the measured volume (p = 0.149). The U-Pflex differed among the trials (p = 0.003) and the methods used for its calculation (p < 0.01). Constant flows from 1 to 10 L/min can equally determine L-Pflex in ARDS patients and is an easy and quick way to obtain the L Pflex in order to optimize positive end expiratory pressure (PEEP) in ARDS patients. PMID- 11903140 TI - Reciprocal influences between ambulatorial peritoneal dialysis and pulmonary function. AB - The aim of this study was to verify if dialysis solution volumes used in patients on continuous ambulatory peritoneal dialysis (CAPD) interfere with pulmonary function and if the pulmonary volumes interfere with the lymphatic absorption of the peritoneal cavity. We submitted 10 CAPD patients with a mean age of 48 +/- 18 years and on CAPD for 35 +/- 27 months to the following evaluations: first, measurement of the lymphatic absorption from the peritoneal cavity; second, measurement of the hydrostatic intraperitoneal pressure; and third, expirometry with the peritoneal cavity full of dialysis solution and empty. There were no differences between the expirometry results obtained with the peritoneal cavity full and empty of dialysis solution, and the results were in accordance with the prediction for this population. The values did not correlate with the peritoneal lymphatic absorption of the peritoneal cavity. The cumulative lymphatic absorption of the peritoneal cavity after 4 h dialysis solution permanence was 197 +/- 93 ml, and the hydrostatic intraperitoneal pressure was 13.9 +/- 2.8 column centimeters of water. Neither of these correlated with pulmonary volumes. In conclusion, CAPD did not interfere with the pulmonary function, nor did the pulmonary function influence the lymphatic absorption of the peritoneal cavity of these patients. PMID- 11903142 TI - Real time monitoring of oxygen saturation in extracorporeal circulation using an optical reflectance transducer. AB - A novel optical reflectance transducer for continuous and real time monitoring of oxygen saturation (So2) in extracorporeal tubings during cardiopulmonary bypass (CPB) is described. The optical transducer integrates 2 light-emitting diodes and a photodiode. The transducer is attached to the extracorporeal tubing by a transparent flow through a 3/8 x 3/8 inch connector/cuvette. Four transducers were built and evaluated in vitro in a mock extracorporeal system at a controlled So2. The correlation index (r2) between the So2 measured by the transducers and values determined by a commercial blood gas analyzer was 0.998 with an absolute difference < 0.5%. The performance of the transducer was also evaluated during 16 CPB surgeries in patients with an r2 of 0.873 and an absolute difference < 3.5%. The results obtained demonstrate the applicability of the optical reflectance transducer for monitoring So2 in the tubings of the CPB circuit. PMID- 11903143 TI - Clinical experience with heart valve homografts in Brazil. AB - The purpose of this paper is to describe the development and progress of the first heart valve homograft bank in Brazil as well as to report the 5-year clinical results. The bank was started in 1995 and employs modern techniques of cryopreservation. Organ procurement increased from 11 hearts in 1995 to 138 hearts in 2000. In the beginning of the experience, only 2 hospitals were using these valves, but this increased to 18 centers in 2000. Clinical experience at the major center includes 117 cases of the Ross procedure, 62 aortic homograft implantations, and 18 cases of mitral homografts. Five-year survival after the Ross procedure was 99.1%, and survival free from any kind of complication was 88.8%. No patients are on anticoagulants, and the incidence of thromboembolism was null. We conclude that auto- and homografts are probably the best alternative to aortic valve replacement for young patients in developing countries. PMID- 11903144 TI - In vitro construction of a potential skin substitute through direct human keratinocyte plating onto decellularized glycerol-preserved allodermis. AB - This work demonstrates that glycerol-preserved acellular allodermis can be used as support for the proliferation of human keratinocytes and that the characteristics of this bioengineered tissue suggest its possible use as a permanent skin substitute for therapeutic challenges such as extensive burns as well as its possible use as an in vitro model for pharmacological studies. The removal of all basal membrane components during preparation of the dermal support also provides an original in vitro situation that allows observation of the reorganization of the dermal-epidermal junction. The tissue composite obtained is constituted of dermis covered by a well attached, multistratified epithelium with morphological characteristics that resemble human epidermis as evidenced by light and transmission electron microscopy, including the neoformation, albeit incomplete, of the dermal-epidermal junction. Assessment of involucrin and cytokeratin 14 expression by immunohistochemical assays established differentiation patterns. Both immerse and air-liquid interface culture systems were tested. PMID- 11903145 TI - Pulsatile ventricular assist device with pericardial inner lining. AB - Preserved pericardium in contact with blood is not thrombogenic, therefore avoiding the use of anticoagulants, and has excellent mechanical properties. Our objective is to take advantage of these characteristics and build a pulsatile ventricular assist device (VAD) with pericardium used as the inner lining of the blood chamber. A mold is used for the tanning of the pericardium, rendering it with an exact shape. A flexible polymeric structure is designed to serve as a base for the pericardium, guiding it and limiting its rate of strain. It consists of two halves, which when outfitted with the interior pericardium lining and connected to each other, form the blood chamber. This assembly is housed in rigid polyvinyl chloride (PVC) shells making up the air chamber for the pneumatic activation. Valves are likewise made of pericardium. Sealing of the chambers was tested statically up to 300 mm Hg with no air or fluid leakage. The device was tested for 60 continuous days in a mock loop, demonstrating hydrodynamic performance adequate for ventricular assist. Micrographs (confocal laser and scanning electron microscopy) were obtained of several pericardium areas, especially on the flexing regions that are a transition between the wet and dry regions. No sign of damage to the pericardium was observed either with the naked eye or at the microscopic level. From the hydraulic performance and materials viewpoints, a completely pericardium-lined pulsatile VAD displaying a polymeric structure that avoids unpredictable bending and limits strain is feasible. The results warrant further studies regarding biocompatibility and strength advantages. PMID- 11903146 TI - Mechanical behavior and stability of the internal membrane of the InCor ventricular assist device. AB - This paper describes and analyzes the mechanical behavior of the internal membrane of the InCor VAD (Heart Institute [InCor], University of Sao Paulo, Brazil), applying the knowledge and tools of structural engineering analysis. This membrane plays an important role in the operation of the ventricular assist device (VAD) because it separates the blood chamber from the pneumatic one, transmitting the pneumatic load to the blood, thus making the desired blood flow possible. The loading repeats itself every time the VAD beats. Therefore the performance, reliability, and durability of the membrane are critical for the performance of the VAD. The mathematical model is based on the large deflection theory of thin shells and on the finite element method. The snap-through instability phenomenon, which is responsible for transmission of the pneumatic load to the blood, was observed in the membrane both when modeled mathematically and experimentally. Principal stresses and strain distributions were obtained with this model at certain load levels along the pre- and postbuckling paths. PMID- 11903147 TI - Initial management of severe hemorrhage with an oxygen-carrying hypertonic saline solution. AB - We tested the hypothesis that the combination of polymerized bovine hemoglobin (PBHg) with hypertonic saline may be beneficial for the initial management of hemorrhagic shock in 22 mongrel dogs (15 +/- 1 kg) bled to a mean arterial pressure (MAP) of 40 mm Hg in 5 min and maintained at this level for 45 min (shed blood volume approximately 50 ml/kg). Animals were treated with a 4 ml/kg bolus over 4 min of one of the following fluids: whole blood, 7.5% NaCl (HS), 13 g/dl of PBHg, or 7.5% NaCl combined with polymerized bovine hemoglobin (HS-PBHg). No additional intervention was performed, and the animals were followed for 60 min after treatment. PBHg and HS-PBHg produced a sustained, significant increase in MAP. Cardiac output was transiently increased only after HS and HS-PBHg. A partial increase in superior mesenteric artery blood flow was observed, particularly after HS-PBHg. We concluded that small volumes of PBHg alone restore MAP, but not blood flow. The combination of PBHg with hypertonic saline provides improvements in cardiac output and mesenteric blood flow, suggesting a potential benefit for the initial management of major blood loss. PMID- 11903148 TI - Effects of hemoglobin-based blood substitutes on vasoactivity of rat aortic rings. AB - Our objective is to characterize the vasoactive properties of a 10% alphaalpha diaspirin cross-linked human hemoglobin (alphaalphaHb) and to test the hypothesis that sodium nitroprusside (SNP)-induced relaxation is inhibited in the presence of alphaalphaHb. Experiments were performed on aortic rings from 18 Wistar rats; the rings were suspended in aerated Krebs solution. Changes in isometric tension were measured to increasing concentrations of alphaalphaHb (1.8 x 10(-9) to 10( 4) M) on phenylephrine (PE)-induced contraction (3 x 10(-7) M), on acetylcholine (ACh)-induced relaxation (10(-8) to 10(-6) M), on SNP-induced relaxation (10(-9) and 10(-8) M), and on PE-induced contraction with an endothelin-1 (ET1) receptor antagonist, BQ123 (10(-5) M). Control rings received no alphaalphaHb. A concentration-dependent increase of the PE-precontraction (1.3%, 6.8%, 17.4%, and 34%, respectively) as well as the inhibition and reversal of ACh-induced relaxation was observed after alphaalphaHb. The presence of alphaalphaHb decreased the SNP-induced relaxation in the presence or absence of endothelium. The relaxation induced by SNP was reduced with time in the presence, but not in the absence, of alphaalphaHb. In conclusion, although pharmacological modulation of the vasoconstriction is possible with nitric oxide donors, our findings suggest that in the clinical setting, large sustained donor doses may be required. PMID- 11903149 TI - A new approach to the study of latissimus dorsi muscle vasoreactivity in rats. AB - In this paper we describe a new approach to the study of changes in latissimus dorsi (LD) muscle microcirculation in rats. The experiments were carried out under anesthesia in normal male Wistar rats (C, n = 6) and in diabetes-induced rats (D, streptozotocin, 50 mg/kg, i.v., n = 6). The left LD muscle was exposed in order to preserve the proximal tendon with its thoracodorsal nerve and artery. The animal was kept in lateral decubitus over a heating board attached to the mechanical stage of the intravital microscope. The ventral surface of the muscle was exposed over a transparent plate and fixing. The image of the LD vascularization was transferred to the camera system, which was connected to a microcomputer equipped with software (KS-300, Kontron Elektronik, Munich, Germany) for image storage. The vasoreactivity of LD was analyzed by changes in arteriole diameter after topically administered noradrenaline (0.3 microg/ml) and acetylcholine (300 microg/ml). The microscopic image provided by the described optical setup permitted clear resolution of capillary vessels and a stable preparation over a period of 3-4 h. D rats showed increased vasodilatation (29 +/ 2% vs. 18 +/- 2.6% in C) and similar vasoconstriction (25.5 +/- 3% vs. 27.5 +/- 3.3% in C) as compared to C rats. The method described in this paper is suitable for the study of changes in responsiveness of LD arterioles, vessels which represent the major site of vascular resistance and are most actively involved in the control of tissue perfusion. PMID- 11903150 TI - Nonstimulated cardiomyoplasty improves hemodynamics in myocardial-infarcted rats. AB - Cardiomyoplasty has been proposed as an alternative surgical treatment for congestive heart failure. The girdling effect of the muscle wrap is believed to reduce diastolic wall stress. We tested the hypothesis that nonstimulated or passive cardiomyoplasty (CDM) would reduce hemodynamic deficits in rats with experimentally induced myocardial infarction (MI). Four groups of animals were studied: intact (C, n = 6), CDM (n = 6), MI by ligation of the left coronary artery (n = 6), and left latissimus dorsi CDM performed 14 days post-MI (MI + CDM, n = 6). All groups were studied 8 weeks after MI and/or CDM or from the beginning of the experiment in controls. MI rats had a lower mean arterial pressure and higher end-diastolic pressure (EDP) compared with controls. End diastolic pressure (EDP) and the left ventricular-body weight ratio (LV/BW) were reduced in the MI group after CDM. These data suggest that passive girdling of the heart provided by CDM may improve post-MI cardiac function. PMID- 11903151 TI - Vulval disease in pre-pubertal girls. AB - Children present with vulval complaints less frequently than do adults; although there are many similarities between paediatric and adult groups of patients with vulval disease, there are also important differences. In both groups, dermatitis, psoriasis and lichen sclerosus are the most frequently seen dermatoses. Birthmarks and congenital abnormalities presenting for the first time are more of an issue in children than in adults. Fusion of the labia and streptococcal vulvovaginitis are conditions seen only in the paediatric group. Sexually transmitted diseases such as genital warts and genital herpes are not common in this group and should always raise the possibility of child sexual abuse. Chronic vulvovaginal candidiasis, although a very common problem in adult patients, is not seen in the prepubertal group. PMID- 11903153 TI - Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis is a rare condition that can affect any organ of the body. Patients of all ages may present to the dermatologist and it is important to make the diagnosis as quickly as possible, because time from presentation to diagnosis is of prognostic importance in adults with the disease. Langerhans cell histiocytosis is now classified as a class I histiocytosis and research into this disease has been very active over the past 10 years. We now know that the disease represents a clonal expansion of a Langerhans cell, which shows phenotypic evidence of activation. The tissue damage induced by the disease appears to be related to local cytokine release. In single system disease, Langerhans cell histiocytosis is responsive to local therapy but, in resistant single system disease or in multisystem disease, etoposide is the most effective monochemotherapy. Some patients will need maintenance treatment with azathioprine or 6-mercaptopurine with or without methotrexate. In such cases, physicians who are used to treating chronic and relapsing diseases and who have experience with these drugs, such as dermatologists, are the most appropriate to manage patients with Langerhans cell histiocytosis. PMID- 11903154 TI - Objective assessment of port-wine stains following treatment with the 585 nm pulsed dye laser. AB - Previous studies assessing the treatment of port-wine stains (PWS) with the 585 nm pulsed dye laser have relied on either subjective clinical assessment or in vivo measurement of skin colour alone. The aim of the present retrospective study was to develop an objective method of assessing available pre- and post-treatment photograph pairs. Port-wine stains depicted in photographs of 23 patients following six or more treatment sessions were assessed for changes in colour (DeltaH*) and PWS size by computer image analysis and were compared with a subjective assessment of PWS reduction by a "blinded" physician examining the same images. The post-treatment mean reduction in the PWS assessed by the physician was 39.7%. A global assessment score incorporating values of DeltaH* and PWS size by computer analysis showed a mean reduction of 12%, with a more significant correlation with the physician assessment (Spearman's rank correlation coefficient 0.627; P=0.001) than changes in size or colour alone. PMID- 11903155 TI - Accuracy and reliability of store-and-forward teledermatology: preliminary results from the St George Teledermatology Project. AB - Teledermatology is the practice of dermatology across distances (and time) and involves the transfer of electronic information. To be effective and safe, the teledermatology process needs to demonstrate an acceptable level of accuracy and reliability. Accuracy is reflected by the degree of concordance (agreement) between the teledermatology and face-to-face diagnoses. Reliability is dependent on how consistently a set of results can be reproduced across different operators. Mean concordance (primary diagnoses) achieved by four dermatologists studying 53 store-and-forward diagnostic cases, originating from 49 referred patients, was 79% (range 73-85%). When the differential diagnoses were taken into account, the variation across individual dermatologists narrowed further, with a mean of 86% (range 83-89%). In contrast, the mean general practitioner (GP; n=11) concordance (GP face-to-face vs reference dermatologist store-and-forward diagnoses) was 49%. An interim review of all 49 teledermatology patients showed no adverse outcome at the end of 3 months. The ability to request face-to-face visits by dermatologists, combined with GPs maintaining primary care of the referred patient, serve as additional safeguards for patients using a telemedicine system. Our results indicate that teledermatology management of referred skin complaints is both accurate and reliable. PMID- 11903156 TI - Inflammatory linear verrucous epidermal naevi: a review of 23 cases. AB - Twenty-three patients diagnosed with inflammatory linear verrucous epidermal naevi (ILVEN) over a 13 year period are reviewed retrospectively. These ILVEN showed a predilection for the buttock and legs and were usually unilateral. Onset of the lesions was usually within the first 6 months of life and extension beyond the original margins occurred in 26% of cases. Sixteen of the 23 patients were male. Ten patients had ILVEN predominantly involving the left side, 12 were on the right side and, in one case, the side involved was not recorded. PMID- 11903157 TI - Report of 19 cases of photoallergic contact dermatitis to sunscreens seen at the Skin and Cancer Foundation. AB - We report on our experience with sunscreen allergy between 1992 and 1999 and also review the international literature on sunscreen allergy. There were a total of 21 allergic reactions to sunscreen chemicals observed in 19 patients over the 8 years. There were nine positive photopatch reactions to oxybenzone, eight to butyl methoxy dibenzoylmethane, three to methoxycinnamate and one to benzophenone. No positive reactions were observed to para aminobenzoic acid. Six patients also had positive patch tests to components of the sunscreen base. In our experience, sunscreen chemicals are the most common cause of photoallergic contact dermatitis. PMID- 11903158 TI - Tinea due to Trichophyton violaceum and Trichophyton soudanense in Hamilton, New Zealand. AB - Between 1994 and 2000, 63 isolates of Trichophyton violaceum and five isolates of Trichophyton soudanense were recorded in both private and public laboratories in Hamilton, New Zealand. A retrospective analysis of medical records of these patients was performed. From these 68 isolates, 58 were recovered from scalp specimens and 10 were recovered from other body sites. There were 51 patients with tinea capitis and nine patients in the tinea corporis group. Six patients had more than one isolate reported at different laboratories. As expected, the vast majority of scalp infections (46/51 patients) were children, with an overall median age of 6 years (range 8 months to 66 years). All patients in the tinea capitis group, except one, were refugee immigrants from East Africa. Of nine patients in the tinea corporis group, six were refugees from the same area. For tinea capitis, 31 patients received systemic antifungal therapy for at least 4 weeks, with either terbinafine (21 patients), griseofulvin (four patients) or itraconazole (six patients). Five patients received topical antifungal creams or shampoo as monotherapy only. The remainder (15 patients) received either no therapy or no record was available. The emergence of these two pathogens as causes of tinea capitis in Hamilton closely correlates with the increasing number of refugees from endemic areas. There is a high rate of person-to-person transmission with these anthropophilic organisms in children as well as adults in the family. Transmission of infection to the local population has been observed, but there is no evidence to date to suggest that these organisms have become endemic in the local population. PMID- 11903159 TI - Artefactual skin disease in children and adolescents. AB - As in adults, artefactual skin disease in children and adolescents has heterogenous presentations with multifactorial aetiology. We report a series of 32 young patients aged 8-16 years. There were 24 females and 8 males. In over half of the cases the lesions were on the head and neck. The types of lesions encountered included physical injury producing grazing, erosions and deep ulcers, chemical and thermal burns, hair cutting and shaving and skin painting. We discuss the approach that we believe should be taken with these patients, emphasizing the role of dermatology-psychiatry liaison in their management. Of great importance is the avoidance of confrontation of the patient but the clear exposition of the nature of the problem to the parents. The major aim should be to have the family accept the need for expert psychiatric assistance. PMID- 11903160 TI - Antiepiligrin (laminin 5) cicatricial pemphigoid complicated and exacerbated by herpes simplex virus type 2 infection. AB - A 50-year-old man with antiepiligrin (laminin 5) cicatricial pemphigoid (AeCP) involving the eyes, mouth and skin required a combination of systemic drug therapies to suppress the ocular disease. Herpes simplex virus type 2 infection of the mouth and pharynx precipitated an acute deterioration, with laryngeal involvement and an increase in oral ulceration. This is an unusual complication of long-term immunosuppression and illustrates some of the difficulties in the management of patients with AeCP. Clinical improvement was obtained with oral antiviral therapy and adjustment of his immunosuppressive regimen. PMID- 11903161 TI - Lupus vulgaris in a child following BCG immunization. AB - A three year old girl presented with lupus vulgaris of the upper arm, which appeared 1 month after BCG immunization. The diagnosis was confirmed by polymerase chain reaction after histology and culture were negative for mycobacteria. Complete healing followed 6 months of oral isoniazid. PMID- 11903162 TI - Bullous lesions in Bazex syndrome and successful treatment with oral psoralen phototherapy. AB - A 59-year-old man presented with a psoriasiform dermatitis with associated bullae and destructive nail dystrophy of the hands and feet. He had lost 10 kg weight over 6 months and a mass in the neck was noted. He was provisionally diagnosed with Bazex syndrome (acrokeratosis paraneoplastica) and subsequent investigations revealed a squamous cell carcinoma in the right piriform fossa. His skin lesions were treated with oral psoralen and ultraviolet A phototherapy and this cleared the cutaneous changes, but the nail changes have persisted at 18 months follow up. PMID- 11903163 TI - Long-term thalidomide for actinic prurigo. AB - A 35-year-old man presented at the age of 8 years with recurrent pruritic papulovesicular lesions on his face and body appearing within minutes of light exposure. A recent positive finding of human leukocyte antigen (HLA) DR4 with the rare DRB1*0407 subtype confirmed a diagnosis of actinic prurigo. Thalidomide (100 mg/day) was commenced at the age of 11 years after an unsuccessful trial of other treatments and his lesions resolved within 2 months. Attempts to withdraw thalidomide have resulted in recurrence of photosensitivity and the patient has remained on a virtually continuous maintenance dose of thalidomide (50 mg/ day) for 23 years. His cumulative dose is estimated to be over 400 g. To date, he has not experienced any adverse effects and investigations have shown no evidence of neuropathy. This case illustrates the safe long-term use of thalidomide. PMID- 11903164 TI - Reactive perforating collagenosis: a condition that may be underdiagnosed. AB - Reactive perforating collagenosis is a perforating disorder developing in adults, usually in association with diabetes mellitus or renal failure. We present three cases diagnosed at the Royal Prince Alfred Hospital in a 5 month period. All three patients had long-standing diabetes mellitus, hypertension, hypercholesterolaemia and ischaemic heart disease. Each patient presented with generalized pruritus and a papular eruption across the trunk and limbs. More than one biopsy or multiple levels were needed before the diagnostic histological features were seen. The first patient responded to 0.5% phenol with 10% glycerine in sorbolene cream. The second patient did not respond to topical betamethasone diproprionate 0.5 mg/g cream and antihistamines (hydroxyzine 25 mg nocte) and required narrow-band ultraviolet (UV) B. The third patient, having failed to respond to topical betamethasone diproprionate 0.5 mg/g cream and wet dressings, antihistamines (hydroxyzine 25 mg tds and doxepin 50 mg nocte) and UVB required acitretin 25 mg orally per day. Because reactive perforating collagenosis responds to treatment, we believe this condition should be considered in patients with diabetes mellitus or renal failure presenting with pruritus and that biopsy of intact lesions may need multiple levels to help establish the diagnosis. PMID- 11903165 TI - Collarette scaling in pityriasis rosea demonstrated by digital epiluminescence dermatoscopy. AB - Collarette scaling is a characteristic sign in pityriasis rosea. The use of digital epiluminescence dermatoscopy is proposed to assist in the recognition of this sign as this technique can magnify the lesions, eliminate other epidermal changes, and demonstrate the morphology and direction of scaling. PMID- 11903166 TI - Sarcoidosis with prominent giant cells. AB - A 31-year-old man presented with a widespread papular eruption and systemic symptoms including renal colic and decreased exercise tolerance. The combination of clinical features and laboratory investigations that revealed an elevated angiotensin converting enzyme level and hypercalcaemia enabled a diagnosis of sarcoidosis to be made. Multiple skin biopsies showed prominent Touton-like giant cells which delayed the diagnosis. Giant cells are frequently seen in sarcoidal granulomas but in some cases their prominence and Touton-like appearance may suggest alternative diagnoses such as xanthogranulomas. PMID- 11903167 TI - Angiokeratoma circumscriptum naeviforme: rare presentation on the neck. PMID- 11903168 TI - Multiple corticosteroid allergies. PMID- 11903170 TI - Various diffuse infiltrative lung diseases: high resolution computed tomography appearances. AB - High resolution CT (HRCT; 1-2 mm collimation scans reconstructed with a high spatial frequency algorithm) is an established technique for the evaluation of diffuse infiltrative lung diseases. For many of these diseases, the features shown on HRCT correlate well with the histopathological abnormalities. The purpose of this pictorial essay is to illustrate HRCT appearances in various diffuse infiltrative lung diseases. PMID- 11903171 TI - Magnetic resonance imaging of the wrist ligaments. AB - The advent of small arthroscopes has enabled the hand surgeon to visualize the ligaments of the wrist directly with resultant increased accuracy in diagnosing and treating pathology. Orthopaedists are now demanding a preoperative assessment and this, in turn, has necessitated that radiologists have a comprehensive understanding of wrist anatomy. High resolution MR imaging can identify the wrist ligaments reliably and provide information concerning their integrity. Interpretation is not straightforward; there is considerable anatomic variation, and there are perforations, defects and degenerative tears that can be troublesome in diagnosing injury. However, with experience and attention to anatomic detail, the radiologist can provide useful information regarding structural abnormalities. When injured, the ligaments of the wrist behave as other joint ligaments do. Findings following injury include discontinuity of normal striated bands, incomplete disruption, irregularities and alteration in normal signal. Fluid pooling around a ligament and concomitant bone injury are other clues to injury. The identification of such structural abnormalities may help to explain altered biomechanics and improve the management of patients following wrist injury. PMID- 11903172 TI - Intramuscular fluid collections and their association with longitudinal rotator cuff tears. AB - This study draws attention to the association between intramuscular fluid collections occurring at the myo-tendinous junction of the rotator cuff secondary to longitudinal tears of their tendons. PMID- 11903173 TI - Chordoma: review of clinicoradiological features and factors affecting survival. AB - This study reviews the clinicoradiological features of cranial and sacrospinal chordomas and identifies factors affecting survival. Nineteen patients seen between January 1980 and December 2000 with histopathological diagnosis of chordomas were retrospectively reviewed with reference to clinical presentation, imaging features, treatment modalities and post-therapy status. Eight had tumours in the skull base while 11 patients had spinal and sacrococcygeal lesions. Surgical resection was performed in 16 patients whose subsequent natural history was used to identify clinical indicators that may influence survival. Completeness of resection, age, gender and postoperative irradiation were subjected to analysis using the Cox proportional hazard models. Kaplan-Meir survival curves illustrate the survival distributions. Diplopia and facial pain are prime clinical presentations in cranial lesions, while extremity weakness and a sacrogluteal mass are common complaints in the sacrospinal group. Lesional calcifications are present in 40% while an osteolytic soft tissue mass is detectable by CT in all cases. Heterogeneous signals and internal septations on T2-weighted MRI are predominant features. In sacrospinal tumours, complete excision with adjuvant radiotherapy achieves the best results with a disease-free survival of more than 5 years. The clinical and imaging findings in this study are in accordance with those of other series. Except for complete surgical excision followed by radiotherapy in the subset of patients with sacrospinal tumours, none of the other clinical indicators show a statistical significant influence on survival. PMID- 11903174 TI - Disappearing liver sign. AB - The development of a pneumothorax is a recognized complication of ultrasound guided percutaneous liver biopsy. A way of detecting such a complication using real-time ultrasound is described, whereby the well-visualized liver disappears from the monitor screen. We also discuss the reasons for this occurrence. PMID- 11903176 TI - Efficacy of daily bedside chest radiography as visualized by digital luminescence radiography. AB - To determine the diagnostic impact of daily bedside chest radiography in comparison with digital luminescence technique (DLR; storage phosphor radiography) and conventional film screen radiography, a prospective randomized study was completed in 210 mechanically ventilated patients with a total of 420 analysed radiographs. The patients were allocated to two groups: 150 patients underwent DLR, and 60 patients underwent conventional film screen radiography. Radiological analysis was performed consensually and therapeutic efficacy was assessed by the clinicians. There was no statistical significant difference between the frequency of abnormal findings seen on DLR and conventional film screen radiography. In total, 448 abnormal findings were present in 249 of 300 DLR and 97 of 120 conventional film screen radiographs. The most common findings were signs of overhydration (41%), pleural effusion (31%), partial collapse of the lung (11%) and pneumothorax (2%). One hundred and twenty-three of 448 (27%) of these abnormal findings were thought to have a considerable impact on patient management. The high rate of abnormal findings with significant impact on patient management suggests that the use of daily bedside chest radiography may be reasonable. PMID- 11903177 TI - Primary leiomyosarcoma of inferior vena cava, a rare entity: Imaging features. AB - Leiomyosarcoma of the inferior vena cava (IVC) is an uncommon neoplasm, most frequently seen in the sixth decade with a female predominance. Imaging modalities allow an early and accurate preoperative diagnosis resulting in a higher rate of surgical resection and improved survival. Imaging findings in a 65 year-old woman with leiomyosarcoma of IVC are described. Computed tomography and MRI typically showed a non-fatty, necrotic intraluminal IVC mass with extension to retroperitoneal compartments. Cavography was useful to evaluate the collateral circulation associated with the extensive intraluminal mass. The lesion was biopsied under ultrasound guidance. PMID- 11903178 TI - Analysis of a clinical sign in traditional Chinese medicine using Doppler ultrasound. AB - Using Doppler ultrasound, a study was done on the characteristics of the radial pulse at the wrist in pregnant and non-pregnant women in order to test the hypothesis that the traditional Chinese medicine 'smooth pulse' of pregnancy has a physiological basis. Comparison of pulse characteristics in a small series of subjects strongly supports this hypothesis. PMID- 11903175 TI - Treatment of renovascular disease with percutaneous stent insertion: long-term outcomes. AB - Renal artery stenosis is a common, progressive cause of hypertension and renal impairment, and is frequently treated with percutaneous transluminal dilatation and stenting. The outcome of this procedure is still being evaluated. The records of 198 consecutive patients who had stents inserted at the Royal Melbourne Hospital were analysed retrospectively, and adequate follow-up information on 148 (75%), in whom a total of 182 renal arteries had been treated was obtained. Technical success was achieved in 144 patients (97%). Complications occurred in 19 patients (13.3%), with major complications occurring in 10 (7.0%) and one death occurring in relation to the procedure. A fall in average systolic blood pressure of 13.2 mmHg (12.1-14.3 mmHg) was seen and a fall in diastolic blood pressure of 10.1 mmHg (9.3-10.9 mmHg), without an increase in the number of antihypertensive drugs used. Renal function remained stable in the majority of patients, particularly those who had minimal baseline renal impairment. Restenosis was common after 6 months, occurring eventually in 29% of screened patients, but was not shown to affect clinical outcomes. Insertion of renal artery stents is a safe and effective treatment for renal artery stenosis. PMID- 11903180 TI - Comparison of chronic exposures received by radiation workers using different biological end-points with the doses recorded by TLD. AB - The frequency of different biological end-points such as translocation, dicentrics (DC) and micronuclei (MN) was studied in 14 radiation workers and 21 non-radiation workers. The average frequencies for different types of aberrations were significantly higher in radiation workers compared to those of respective aberrations in non-radiation workers. Out of 14 radiation workers, eight subjects showed a dose above the detection limit as per translocation and seven subjects as per DC frequency and no patient showed a dose above the detection limit as per MN frequency. Regression analysis carried out between the recorded doses according to Thermo Luminescence Dosimeter (TLD) and the dose estimated as per translocation frequency gave a correlation coefficient of 0.32, whereas that obtained with TLD dose and the dose estimated as per DC was 0.81. When the correlation was made between the TLD dose, which was above 0.15 Gy (the detection limit for translocation), and the dose estimated as per translocation frequency in these subjects, a correlation coefficient of 0.98 was found. A similar analysis between the TLD dose above 0.5 Gy (the detection limit for DC) and the dose estimated as per DC frequency in these subjects, a correlation coefficient of 0.26 resulted. This paper discusses the reasons for the poor correlation obtained between TLD dose and dose estimated as per DC and MN frequency. PMID- 11903179 TI - Phase III, randomized, double-blind, cross-over comparison of gadoteridol and gadopentetate dimeglumine in magnetic resonance imaging of patients with intracranial lesions. AB - This study compares the efficacy and safety of gadoteridol with that of gadopentetate dimeglumine for enhanced MRI in subjects with intracranial lesions. A total of 92 subjects at three European centres underwent one MRI study enhanced with 0.1 mmol/kg gadoteridol and another with 0.1 mmol/kg gadopentetate dimeglumine. Contrast agents were assigned in random order, separated by 3-7 days. Eighty subjects were evaluated for efficacy. The presence of pathology, degree of enhancement, location and number of lesions, as well as additional information gained, were compared for each subject's unenhanced and enhanced scans for both the gadoteridol and gadopentetate dimeglumine examination. Safety was evaluated in all treated subjects by means of pre- and post-dose vital signs, laboratory tests and by monitoring for adverse events. There was no significant difference in the number of lesions visualized pre- and post-contrast for the two contrast agents. A high degree of correlation was noted between the two blinded readers. When post-contrast image sets were compared between contrast agents, there was no significant difference in superiority of one agent over the other for any of the evaluators (P > 0.05). No significant differences for any safety parameter were noted between the two agents. Gadoteridol and gadopentetate dimeglumine are effective and well tolerated for use in contrast-enhanced MRI of the CNS at a dose of 0.1 mmol/kg. PMID- 11903181 TI - Diagnosis and management of astrocytomas, oligodendrogliomas and mixed gliomas: a review. AB - Low-grade gliomas are a diverse group of neoplasms which, as the name implies, are thought to arise from glial cells. Common among this group are astrocytomas (low-grade astrocytoma; LGA), oligodendrogliomas and mixed gliomas. Among these, LGA is the commonest low-grade glioma and, occasionally, although incorrectly, the terms are used interchangeably. Advances in imaging technology have improved the accuracy of preoperative diagnosis. The management of low-grade gliomas is controversial. Recent evidence suggests that previously considered standard therapy (i.e. surgery plus radiotherapy) may not be in the patient's best interests. A review of the available published research concerning low-grade gliomas is therefore timely. PMID- 11903182 TI - Imaging with F-18 FDG PET is superior to Tl-201 SPECT in the staging of non-small cell lung cancer for radical radiation therapy. AB - Thallium-201 (Tl-201) single photon emission computed tomography (SPECT) is funded for evaluation of malignancy in Australia and may have utility for staging of non-small cell lung cancer (NSCLC) if CT results are equivocal. Fluorine-18 fluorodeoxyglucose (F-18 FDG) positron emission tomography (PET) is superior to CT for staging NSCLC but is more expensive and less widely available than Tl-201 SPECT. Therefore, these techniques were prospectively compared in 27 radical radiation therapy candidates. Patients were allocated a conventional, PET and Tl 201 stage. Tumour to background ratios (TBR) were recorded for the primary on both techniques. Metastatic disease was confirmed by surgical pathology, serial imaging or clinical follow up. Tumour to background ratios were consistently higher for FDG PET than Tl-201 SPECT (P < 0.0001). Positron emission tomography detected all known primary tumours but Tl-201 failed to image four primary tumours (15%). In 10 of 18 cases of discordance between PET and Tl-201 SPECT regarding stage, corroboration was available from pathology or disease progression. Positron emission tomography was shown to have a 100% positive predictive value, including all three patients with PET-detected distant metastases (P=0.002). Results indicate that PET is superior to Tl-201 SPECT scanning in the staging of NSCLC for radical radiation therapy, and that the low sensitivity for detection of local and metastatic disease is likely to limit the clinical impact and cost-effectiveness of this technique despite its lower cost. PMID- 11903183 TI - Locally advanced cervix cancer: chemotherapy prior to definitive surgery or radiotherapy. A single institutional experience. AB - Primary or neoadjuvant chemotherapy prior to definitive local therapy has potential advantages for locally advanced cervix cancer. It can downstage a cancer and allow definitive local therapy to be technically possible (surgery), or potentially more effective (radiotherapy). It can also eradicate subclinical systemic metastases. This report reviews a single institution's experience of neoadjuvant chemotherapy prior to definitive local therapy for cervix cancer over a 13-year period. One hundred and six patients were treated with this intent. The patients were analysed for their response to chemotherapy, treatment received, survival, relapse and toxicity. The chemotherapy was feasible and the majority of patients had a complete or partial response (58.5%). Eight patients did not proceed to local treatment. Forty-six patients had definitive surgery and 52 had definitive radiotherapy. The 5-year overall survival was 27% and the majority of patients died with disease. The first site of relapse was usually in the pelvis (46.2%). Late complications that required ongoing medical therapy (n=6) or surgical intervention (n=2) were recorded in eight patients (7.5%). On univariate analysis stage (P=0.04), tumour size (P=0.01), lymph node status (P=0.003), response to chemotherapy (P=0.045) and treatment (P=0.003) were all significant predictors of survival. On multivariate analysis, tumour size (P < 0.0001) and nodal status (P=0.02) were significant predictors of survival. Despite the impressive responses to chemotherapy of advanced cervix cancer, there is evidence from randomized trials that it does not improve or compromise survival prior to radiotherapy. As its role prior to surgery remains unclear, it should not be used in this setting outside a prospective randomized trial. PMID- 11903184 TI - HASTE MRCP and MRI findings in alveolar echinococcosis of the liver. AB - Alveolar echinococcosis is a rare mass-producing inflammatory process of the liver. Experience with MRI, and particularly magnetic resonance cholangiopancreatography (MRCP), demonstrates that features of this disease are limited. The HASTE (half-Fourier acquisition single-shot turbo spin echo) MRCP and MRI findings of alveolar echinococcosis of the liver are presented in this report. HASTE MRCP was used to define the biliary system and the biliary system mass relationship. It was found that results were comparable with those of invasive techniques such as endoscopic retrograde cholangiopancreatography and percutaneous transhepatic cholangiography. PMID- 11903185 TI - Magnetic resonance imaging features of an infected right atrial myxoma. AB - Myxomas are the most common primary intracardiac tumours, most commonly occurring in the left atrium, and rarely become infected. A search of the published research located only six case reports of infected right atrial myxoma. In this seventh reported case, the MRI findings are presented for the first time. PMID- 11903186 TI - Embolization of proper hepatic artery pseudoaneurysm complicating choledochoscopic laser therapy. AB - Biliary papillomatosis is a rare disease characterized by multiple papillary proliferation of the epithelial cell of the bile duct. Because it has a tendency to be recurrent, the treatment strategy is choledochoscopic laser therapy. A patient with biliary papillomatosis treated by choledochoscopic laser therapy, which was complicated by massive haemobilia and shock, is presented. An intrahepatic artery pseudoaneurysm was diagnosed on angiography. A coexisting occlusion necessitated a superselective embolization of the pseudoaneurysm in order to avoid devascularization of the left lobe of the liver. PMID- 11903188 TI - Unusual extrathyroidal iodine accumulation in a post-ablative I-131 scan. AB - A case of iodine-131 accumulation in the gall bladder following an ablative dose for thyroid carcinoma is presented. PMID- 11903187 TI - Hydatid pulmonary emboli. AB - A case of secondary hydatosis, initially misdiagnosed as pulmonary metastases, is presented. The dissemination of hydatid cysts within the lungs in this case was the consequence of direct rupture of a hepatic hydatid into the inferior vena cava. A brief overview of the pathophysiology of hydatid disease, including a discussion of the types of hydatid rupture (contained, communicating and direct), is presented. PMID- 11903189 TI - Investigations of fatal causes of chest pain: case report and literature review. AB - A case of fatal ascending aortic dissection (AAD) misdiagnosed as pulmonary embolism (PE) despite strong radiological evidence is described. The occurrence of this serious pathology is uncommon. Its prompt diagnosis and treatment are crucial. Anticoagulant therapy for pulmonary embolism should be withheld until acute aortic dissection is excluded definitively. A management approach to optimise the outcome of patients with chest pain in which ascending aortic dissection and/or pulmonary embolism are suspected is presented. PMID- 11903190 TI - Multiple bilateral giant fibroadenomas associated with cyclosporine A therapy in a renal transplant recipient. AB - A 31-year-old woman developed a right breast mass following cyclosporine A therapy after a renal transplant. Several large breast masses continued to grow bilaterally. Mammography and ultrasonography showed features of giant fibroadenomas. The diagnosis was confirmed by biopsy of one of the masses. Awareness of the association between cyclosporine A therapy and fibroadenoma development in renal transplant recipients is highlighted. PMID- 11903191 TI - Pancreatic islet cell tumours presenting as recurrent acute pancreatitis: imaging features in three cases. AB - We present three cases of recurrent pancreatitis that occurred in patients with small islet cell tumours of the pancreas which were obstructing the main pancreatic duct. This is a very uncommon presentation of pancreatic islet cell tumours. The radiological findings in these cases are shown and the implications for imaging of 'idiopathic' relapsing pancreatitis are discussed. PMID- 11903193 TI - Prosthetic devices within radiotherapy fields: planning implications and results. AB - Prosthetic devices made of plastic or silicon are occasionally present in patients requiring radiotherapy (RT). The effect of RT on these devices and the potential implications of their presence on RT planning are relatively unknown. Three examples are presented in which various devices were included in RT fields. In two of the examples in vitro testing of the devices with high single doses of radiation was undertaken. Radiotherapy was given to all patients with high doses received by the devices. Minor adjustments were made to the planning techniques to reduce the doses to the devices. Neither significant complications nor any malfunctions of the devices were noted subsequently. PMID- 11903192 TI - Idiopathic hypereosinophilic syndrome: magnetic resonance imaging findings in endomyocardial fibrosis. AB - Significant eosinophilia and even eosinophilic tissue infiltration has been associated with a variety of clinical disorders including allergic and immunodeficiency states, drug reaction, infection, parasitic infestation and malignancy. Eosinophilia without an underlying aetiology and with multi-organ dysfunction has been designated idiopathic hypereosinophilic syndrome. We report a case of endomyocardial fibrosis with MRI findings. PMID- 11903194 TI - 'Indolent' plasmacytoma. AB - A patient presented with lethargy, nausea and diarrhoea and had a 10-year history of neurological symptoms in his legs. He was found to have renal failure. Investigations demonstrated a longstanding plasmacytoma of the sacrum and progression to myeloma. Such an indolent course for plasmacytoma is rare. A review of the clinical case and management of solid plasmacytomas is presented. PMID- 11903196 TI - Comprehensive evaluation of patients with haematuria on multi-slice computed tomography scanner: protocol design and preliminary observations. AB - An imaging protocol with a multi-slice CT scanner that allows comprehensive assessment of patients with haematuria is described. This protocol allows evaluation of the kidneys, ureters and bladder in a single examination using CT. This approach should streamline the diagnostic work-up of patients with haematuria. PMID- 11903195 TI - Isolated hypoglossal nerve palsy due to skull base metastasis from breast cancer. AB - We describe a 44-year-old woman who presented with an isolated unilateral hypoglossal nerve paralysis caused by a skull base metastasis from breast cancer. The patient had a modified radical mastectomy followed by local radiotherapy and adjuvant chemotherapy. Fourteen months later she presented with difficulty in speaking. Physical examination revealed an isolated left hypoglossal nerve paralysis. The MRI scan showed a mass lesion involving the left occipital condyle extending into hypoglossal canal. PMID- 11903197 TI - The future of radiology. PMID- 11903198 TI - RANZCR Conference registration fees. PMID- 11903199 TI - Re: How many new patients should a radiation oncologist see? PMID- 11903200 TI - Wigg and Morgan: 'New' hope? PMID- 11903201 TI - Re: How do waiting times affect radiation dose fractionation schedules? PMID- 11903206 TI - The collaborative practice model for bipolar disorder: design and implementation in a multi-site randomized controlled trial. AB - Bipolar disorder remains a high morbidity and costly illness in general clinical practice, despite the availability of efficacious medications. This 'efficacy effectiveness gap' may be addressed by better organizing systems of care. One type of intervention is the 'collaborative practice model' which can be defined as an organization of care that a) emphasizes development in the patient of illness management skills, and b) supports provider capability and availability in order to c) engage patients in timely, joint decision-making regarding their illness. This article describes such a collaborative practice model for bipolar disorder, designed to be widely adoptable and sustainable in general clinical practice. The first part of the article describes the theoretical background from which the collaborative practice approach developed, emphasizing its origins in the lithium clinics of the 1970s, in nursing theory and practice, and more recently in the management of chronic medical diseases. The second part describes the structure of one such intervention, the Bipolar Disorders Program (BDP) developed in the Veterans Affairs health care system. The third part summarizes results from single-site studies of the intervention. The fourth part describes several key issues in its implementation in an ongoing multi-site randomized controlled trial, VA Cooperative Study Program (CSP) # 430. Data to date indicate that such collaborative practice interventions may improve important process and intermediate outcome variables for bipolar disorder. The BDP provides an example of a multi-faceted collaborative practice model that can be manualized and implemented across multiple sites in a randomized controlled trial. PMID- 11903207 TI - Response to lithium maintenance treatment in bipolar disorders: comparison of women and men. AB - OBJECTIVES: Possible sex differences in responses to mood-stabilizing treatment remain poorly defined. Since women with bipolar disorder reportedly have more features that may predict a poor prognosis (depression and rapid cycling), we tested the hypothesis that women respond less well to lithium maintenance treatment. METHODS: Clinical characteristics of 360 women and men with DSM-IV bipolar I or II disorder were compared before and during clinical lithium maintenance monotherapy in a mood disorders clinic by preliminary bivariate comparisons, multivariate analysis, and survival analysis of time stable during treatment. RESULTS: Women (n = 229) versus men (n = 131) were: more likely to have type II disorder (1.6 times), 3.2 years older at illness onset, more often depressed-before-manic (1.4 times), considered unipolar depressive 1.9 years longer and started maintenance treatment 5.5 years later. However, women differed little from men before treatment in overall morbidity, average episode frequency and risk of suicide attempts. Contrary to prediction, women showed non significantly superior responses to lithium treatment, and a significant 60% longer median time before a first recurrence during treatment, despite 7% lower average serum lithium concentrations. CONCLUSIONS: Women were diagnosed as bipolar later than men with corresponding delay of lithium maintenance treatment that proved to be at least as effective as in men. PMID- 11903208 TI - Clinical correlates of psychiatric comorbidity in bipolar I patients. AB - OBJECTIVES: To ascertain the clinical implications of psychiatric comorbidity in the course and outcome of bipolar I patients. METHODS: One hundred and twenty nine bipolar I outpatients in remission [Young Mania Rating Scale (Y-MRS) < 7, Hamilton Depression Rating Scale (HDRS) < 9] were assessed by means of the Structured Clinical Interview for DSM-III-R axis I and axis II (SCID-I and SCID II) in order to detect all possible psychiatric comorbid diagnoses. The sample was split according to the presence of psychiatric comorbidity and the groups were compared. RESULTS: Psychiatric comorbidity was detected in 31% of the sample. A higher number of mixed features, depressive episodes and suicide attempts and a predominance of depressive onset amongst comorbid bipolar patients were the most relevant differences between the two groups. CONCLUSIONS: There is an association between depression, suicidality and comorbidity in bipolar I disorder. As comorbidity had a clear relevance in the course and outcome of bipolar illness, this issue should be specifically assessed in clinical practice. PMID- 11903211 TI - Postpartum positioning and attachment education for increasing breastfeeding: a randomized trial. AB - BACKGROUND: Although lactation experts suggest that a correct positioning and attachment technique reduces breastfeeding problems and enhances long-term breastfeeding, evidence from randomized trials is lacking. The objective of this study was to evaluate the effect of postpartum positioning and attachment education on breastfeeding outcomes in first-time mothers. METHOD: A randomized trial was performed in a public hospital in Adelaide, South Australia, where 160 first-time mothers were randomly allocated to receive either structured one-to one education (experimental group) or usual postpartum care (control group) within 24 hours of birth. The primary outcome was breastfeeding at 6 weeks and 3 and 6 months postpartum; other outcomes were nipple pain and trauma in hospital and at 6 weeks and 3 and 6 months, and satisfaction with breastfeeding. RESULTS: No significant differences occurred in breastfeeding rates between the groups at each endpoint, although a trend in the direction of lower rates was seen at each endpoint in the experimental group. This group reported less nipple pain on days 2 (p = 0.004) and 3 (p = 0.04), but this was not sustained on follow-up. No differences were observed in nipple trauma in hospital or in self-reported nipple pain and/or trauma at the three endpoints. Experimental group women were less satisfied with breastfeeding at 3 and 6 months postpartum when using a one-item measure; however, a multiple-item measure showed no significant differences at the three endpoints. CONCLUSIONS: The intervention did not increase breastfeeding duration at any assessment time or demonstrate any differences between the groups on secondary outcomes. The trend toward lower breastfeeding rates in the experimental group suggests a need for a larger trial to evaluate whether or nor postpartum positioning and attachment education may negatively affect breastfeeding. PMID- 11903210 TI - Maternal satisfaction with active management of labor: a randomized controlled trial. AB - BACKGROUND: Active management of labor reduces the length of labor and rate of prolonged labor, but its effect on satisfaction with care, within a randomized controlled trial, has not previously been reported. The study objectives were to establish if a policy of active management of labor affected any aspect of maternal satisfaction, and to determine the independent explanatory variables for satisfaction with labor care in a low-risk nulliparous obstetric population. METHODS: Nulliparous women at National Women's Hospital in Auckland, New Zealand, in spontaneous labor at term with singleton pregnancy, cephalic presentation, and without fetal distress were randomized after the onset of labor to active management (n = 320) or routine care (n = 331). Active management included early amniotomy, two-hourly vaginal assessments, and early use of high dose oxytocin for slow progress in labor. Routine care was not prespecified. Maternal satisfaction with labor care was assessed by postal questionnaire at 6 weeks postpartum. Sensitivity analyses were performed, and logistic regression models were developed to determine independent explanatory variables for satisfaction. RESULTS: Of the 651 women randomized in the trial, 482 (74%) returned the questionnaires. Satisfaction with labor care was high (77%) and did not significantly differ by treatment group. This finding was stable when sensitivity analysis was performed. The first logistic regression model found independent associations between satisfaction and adequate pain relief, one-to-one midwifery care, adequate information and explanations by staff, accurate expectation of length of labor, not having a postpartum hemorrhage, and fewer than three vaginal examinations during labor. The second model found fewer than three vaginal examinations and one-to-one midwifery care as significant explanatory variables for satisfaction with labor care. CONCLUSIONS: Active management did not adversely affect women's satisfaction with labor and delivery care in this trial. Future studies should concentrate on measurement of potential predictors before and during labor. PMID- 11903214 TI - Assessing women's preferences for intrapartum care. AB - BACKGROUND: Recent government reports have recommended involving consumers in the planning of health services. Although satisfaction surveys have traditionally been used, they have several limitations. This paper describes a relatively new method of eliciting consumer preferences that allows respondents (women) to indicate the importance that they attribute to specific aspects of a service. The aim was to explore the feasibility of using a discrete choice experiment to assess the importance to women of different aspects of intrapartum care. METHODS: In this pilot study of 301 women at low obstetric risk, data were collected using an anonymous self-complete questionnaire given to each participant by the midwife at the booking visit. RESULTS: The results of the regression model suggest that respondents prefer maternity units that offer greater continuity of caregiver, more methods of pain relief, continuous fetal heart rate monitoring, a homely appearance, routine involvement of medical staff, and greater involvement for the woman in the decision-making process. Although all attributes were important to women, they were not all of equal importance. For example, if continuity of caregiver were achieved at the expense of decreasing the availability of pain relief then women would be worse off. CONCLUSIONS: The discrete choice experiment appears to be a useful tool in assessing the strength of women's preferences for different aspects of maternity care. Future research should include a qualitative approach to explore in greater depth the processes involved in shaping women's preferences. PMID- 11903213 TI - Position for newborn sleep: associations with parents' perceptions of their nursery experience. AB - BACKGROUND: In the United States, sudden infant death syndrome is the leading cause of death among infants between the ages of 1 and 12 months. Although its etiology is unclear, infants who sleep in the prone or side positions are at increased risk. The objective of this study was to examine the association between the perceptions of inner city parents about teaching and modeling during the postpartum period of infant sleeping position, and their choice of sleeping position for their infants. METHODS: A convenience sample of parents of 100 healthy infants who came for the 2-week well-child visit at an urban primary care center were invited to complete a questionnaire and to report on the position in which infants were placed for sleep. RESULTS: Forty-two percent of parents reported that they usually placed their infants in the supine position for sleep; 26 percent placed their infants to sleep in the prone position at least some of the time. Parents who reported being told by a doctor or a nurse to have their infants sleep in the supine position were more likely to choose that position. Similarly, those who reported seeing their infants placed to sleep exclusively in the supine position in the hospital were also more likely usually to choose that position. Parents who reported that they both were told by a doctor or a nurse to put their infants to sleep in the supine position and reported seeing their infants exclusively placed that way in the nursery were the most likely usually to choose that position for their infants to sleep. CONCLUSIONS: Perceptions by parents of instructions from a doctor or a nurse of the position in which the infants were placed in the nursery were associated with the position parents reported placing their infants to sleep at home. Efforts to promote the supine sleeping position in the inner-city setting should address both practices and education provided to parents in the nursery during the postpartum hospital stay and should be sufficiently powerful to align their perceptions of the postpartum experience with current American Academy of Pediatrics recommendations. PMID- 11903212 TI - Epidural analgesia use as a marker for physician approach to birth: implications for maternal and newborn outcomes. AB - BACKGROUND: Understanding the association between caregiver belief systems and practice patterns is an emerging area of research. We hypothesized an association between a maternity caregiver's belief system and his or her behavior. The study objective was to determine if a family physician's overall approach to maternity care, as measured by average use of epidural analgesia, was associated with maternal and fetal outcomes. METHODS: Retrospective analysis was conducted of the births of three cohorts of 1992 nulliparous, low-risk women attended by 96 family physicians within an 18-month period in the department of family practice at the largest maternity hospital in Canada. Cohorts were based on the physicians' mean use of epidural analgesia for the women. Family physicians attending fewer than 5 births were excluded. The main outcome measures, by physician epidural utilization cohort, were maternal/newborn morbidity, procedure rates, consultation rates, and length of stay. RESULTS: Family physicians were separated into cohorts based on their mean use of epidural analgesia at rates of: low, 0-30 percent (15 physicians, 263 births); medium, 31-50 percent (55 physicians, 1323 births); and high, 51-100 percent (26 physicians, 406 births). After adjustment for maternal age and race, patients of low versus high epidural users were admitted at a later state of cervical dilation (mean 4.0 vs 3.1 cm), received less electronic fetal monitoring (76.4 vs 87.2%) and oxytocin augmentation (12.2 vs 29.8%), sustained fewer malpositions (occiput posterior or transverse) (23.2 vs 34.2%), had fewer cesarean sections (14.0 vs 24.4%), less obstetric consultation (47.9 vs 63.8%), and fewer newborn special care admissions (7.2 vs 12.8%). CONCLUSIONS: In our setting, high use of epidural analgesia is a marker for a style of practice characterized by malpositions leading to dysfunctional labors and higher intervention rates leading, in turn, to excess maternal/newborn morbidity. PMID- 11903216 TI - Hospitalization for early bonding of the genetic mother after a surrogate pregnancy: report of two cases. AB - BACKGROUND: In surrogate pregnancies the genetic parents have little opportunity for early bonding to their infant, either prenatally (in utero) or in the immediate postnatal period. The purpose of this article is to describe a new method for encouraging early parent-infant bonding after surrogate pregnancy by hospitalizing the genetic mother in the maternity ward immediately after the delivery. METHODS: Two genetic mothers were hospitalized in the maternity ward (rooming-in system) at the Rabin Medical Center in Israel immediately after delivery of their infants by surrogate arrangement. The first birth was a singleton pregnancy with vaginal delivery and the second, a twin pregnancy with delivery by cesarean section. The genetic parents were counseled by a social worker from the adoption agency, starting 3 months before the estimated date of delivery. The parents were referred to the hospital social worker before the delivery. To assess attachment, we observed the parents' behavior toward their children during two daily 15-minute periods of free, unstructured interaction. RESULTS: The parents showed good primary caregiving functions and established affective verbal and physical contact with the infants. They began to recognize the infants' needs and temperament, and exhibited an aura of self-confidence during their interactions. All expressed satisfaction with the method at discharge and reported on reduction of their fears about returning home with the infants. CONCLUSIONS: We believe that early hospitalization of the genetic mother in a surrogate delivery may be desirable to establish good and safe early mother infant bonding, and that it should be considered for adoption as regular hospital policy. Further randomized studies with larger samples over the long term are warranted. PMID- 11903215 TI - Postdischarge surveillance after cesarean section. AB - BACKGROUND: Cesarean section is a major surgical procedure with a relatively short hospital stay. A significant rate of surgical site infection after this procedure is missed by standard inpatient surveillance. This study aimed to evaluate a method of postdischarge surveillance and compare results with the incidence of infection before discharge. METHOD: A postdischarge survey was sent on day 30 to 277 women who had delivered by cesarean section during the 12-month study period. A follow-up telephone interview was conducted if the questionnaire had not been returned within 2 weeks, if a diagnosis of infection could not be clearly determined from the participant's responses, or to confirm the diagnosis of infection. If follow-up was not completed, a chart audit was undertaken. RESULTS: A total response rate of 89 percent (247/277) was obtained, and 28 women with a surgical site infection were identified from the survey. Telephone follow up and chart review of patients with possible infection and of nonresponders identified 32 percent more postdischarge infections (14/42). The overall infection rate was 17 percent compared with 2.8 percent at discharge. CONCLUSIONS: Postdischarge surveillance is necessary to determine accurate surgical site infection rates after cesarean section, increase awareness of caregivers about infection control problems, and indicate the need for appropriate follow-up care. Women undergoing a cesarean delivery should be informed of the risk of postdischarge infection and educated about the signs and symptoms of infection. PMID- 11903224 TI - Detailed information of the role of authors and that of their sponsors in scientific research. PMID- 11903217 TI - Commentary: routines in maternity units: are they still appropriate for 2002? PMID- 11903225 TI - Topical 5-aminolaevulinic acid photodynamic therapy for the treatment of skin conditions other than non-melanoma skin cancer. AB - Topical 5-aminolaevulinic acid (ALA) photodynamic therapy (PDT) is used increasingly for superficial non-melanoma skin cancer (NMSC) and dysplasia. However, the relative accumulation of the photosensitizer protoporphyrin IX (PpIX) in diseased tissue is not specific for neoplastic disease, and has been shown after the application of ALA to benign proliferative skin conditions such as viral warts and psoriasis. This review appraises the quality of evidence available for the use of topical ALA-PDT in the treatment of skin conditions other than NMSC. The diseases that have been studied in most detail are recalcitrant viral warts, acne, psoriasis and cutaneous T-cell lymphoma. Publications relating to the treatment of other diseases by topical PDT are restricted to small case series or case reports. The relevant literature will be discussed and the potential for topical PDT in the treatment of several skin diseases is highlighted, although more detailed studies are required to clarify the role of PDT beyond the treatment of NMSC. PMID- 11903226 TI - The humanized SCID mouse model to study HLA class II-linked autoimmunity to desmoglein 3 in pemphigus vulgaris. AB - BACKGROUND: Pemphigus vulgaris (PV) is a severe autoimmmune skin disorder induced by antibodies against desmoglein (Dsg) 3 on epidermal keratinocytes. OBJECTIVES: To establish an active animal model of PV to analyse the T-cell-regulated production of pathogenic antibodies in vivo. METHODS: Immunodeficient SCID mice were injected with peripheral blood lymphocytes (PBL) from an HLA-DRB1*0402/DQ8+ patient with PV or a DRB1*0402/DQ8+ healthy donor, with or without subsequent injections of human Dsg3 or preincubation of PBL with Dsg3. RESULTS: Human immunoglobulins (2.7-18.5 mg mL-1) were detected in all the mice after 8 weeks. Only one of 30 PBL-treated mice developed IgM against Dsg3 and showed intercellular IgM deposits in skin, nostrils and tongue. In contrast, in a previous study, 41% of SCID mice injected with PBL from patients with PV developed anti-Dsg3 antibodies in vivo. CONCLUSIONS: Our inability to reproduce these findings may be due to the transfer of slightly lower numbers of PBL (20 x 10(6) vs. 25-30 x 10(6)). PMID- 11903227 TI - Epidermal barrier lipids in human vernix caseosa: corresponding ceramide pattern in vernix and fetal skin. AB - BACKGROUND: Vernix caseosa is a protective biofilm covering the fetus during the last trimester. Vernix and epidermal barrier lipids (i.e. cholesterol, free fatty acids and ceramides) appear to share protective functions for fetal and neonatal skin. OBJECTIVES: To analyse vernix samples for epidermal barrier lipid content, and to compare lipid profiles of vernix with those of fetal and postnatal epidermis. METHODS: Vernix samples were collected from 21 healthy term neonates. Skin samples were collected from 10 fetuses aborted between gestational week (GW) 16 and 25, nine infants and 11 older children. Lipids were extracted according to standard protocols and analysed by high-performance thin-layer chromatography. RESULTS: Vernix contained 196.5 +/- 70.1 microg barrier lipids mg-1 protein (mean +/- SD). Cholesterol formed the major barrier lipid fraction (52.8%), followed by free fatty acids (27.7%) and ceramides (20.1%). The ceramide composition of vernix resembled that of mid-gestational (GW 23-25) fetal epidermis both qualitatively and quantitatively, while there were major differences from postnatal epidermis. The total epidermal ceramide concentration increased significantly between prenatal and postnatal samples. CONCLUSIONS: The composition pattern of ceramides mirrors that of mid-gestational fetal epidermis. Vernix thus represents a 'homologous' substitute for the immature epidermal barrier in fetal skin. The differential role of individual ceramides in this process remains to be established. PMID- 11903228 TI - Propionibacterium acnes and inflammation in acne; P. acnes has T-cell mitogenic activity. AB - BACKGROUND: Circumstantial evidence suggests that Propionibacterium acnes has a role in the inflammation of acne. This could be effected by antigenic or superantigenic or mitogenic reactions. OBJECTIVES: The purpose of this investigation was to determine whether P. acnes had only antigenic activity or additional superantigenic and mitogenic activity. METHODS: A lymphocyte transformation assay was used to detect responses to a mixture of eight P. acnes whole cell isolates, and their supernatant culture fluids. In order to determine the nature of T-cell reactions to P. acnes cells a mouse-antihuman major histocompatibility complex class II monoclonal antibody was used in the lymphocyte transformation assay to inhibit the antigenic stimulation of lymphocytes. An analysis of the T-cell receptor (TCR) variable region beta (BV) repertoire was undertaken using flow cytometry of the unstimulated and stimulated cells. RESULTS: Peripheral blood mononuclear cells (PBMNC) from adults with no history of acne responded strongly to stationary growth phase cells of P. acnes, less strongly to cells in the exponential growth phase. No response was detected to supernatant culture fluids. PBMNC from five cord blood samples (CBMNC) responded maximally after 3 and 7 days of incubation with stationary growth phase cells of P. acnes. The reaction of CBMNC to P. acnes cells was not suppressed completely by the blocking antibody. The analysis of the TCRBV repertoire indicated that P. acnes induced no deletion or over-representation of certain BV element-bearing T cells. The TCRBV analysis was repeated after preincubation with the blocking antibody. Deletion of T cells bearing certain BV components occurred and there was no over-representation of T cells carrying certain BV components. CONCLUSIONS: Two mechanisms of lymphocyte activation by P. acnes cells are proposed, antigen and mitogen driven. These results are consistent with the histological evidence of inflammation in acne lesions. PMID- 11903229 TI - Human Ro60 (SSA2) genomic organization and sequence alterations, examined in cutaneous lupus erythematosus. AB - BACKGROUND: The Ro 60 kDa protein (Ro60 or SSA2) is the major component of the Ro ribonucleoprotein (Ro RNP) complex, to which an immune response is a specific feature of several autoimmune diseases. The genomic organization and any sequence variation within the DNA encoding Ro60 are unknown. OBJECTIVES: To characterize the Ro60 gene structure and to assess whether any sequence alterations might be associated with serum anti-Ro antibody in subacute cutaneous lupus erythematosus (SCLE), thus potentially providing new insight into disease pathogenesis. METHODS: The cDNA sequence for Ro60 was obtained from the NCBI database and used for a BLAST search for a clone containing the entire genomic sequence. The intron exon borders were confirmed by designing intronic primer pairs to flank each exon, which were then used to amplify genomic DNA for automated sequencing from 36 caucasian patients with SCLE (anti-Ro positive) and 49 with discoid LE (DLE, anti-Ro negative), in addition to 36 healthy caucasian controls. RESULTS: Heteroduplex analysis of polymerase chain reaction (PCR) products from patients and controls spanning all Ro60 exons (1-8) revealed a common bandshift in the PCR products spanning exon 7. Sequencing of the corresponding PCR products demonstrated an A > G substitution at nucleotide position 1318-7, within the consensus acceptor splice site of exon 7 (GenBank XM001901). The allele frequencies were major allele A (0.71) and minor allele G (0.29) in 72 control chromosomes, with no significant differences found between SCLE patients, DLE patients and controls. CONCLUSIONS: The genomic organization of the DNA encoding the Ro60 protein is described, including a common polymorphism within the consensus acceptor splice site of exon 7. Our delineation of a strategy for the genomic amplification of Ro60 forms a basis for further examination of the pathological functions of the Ro RNP in autoimmune disease. PMID- 11903230 TI - EEC (Ectrodactyly, Ectodermal dysplasia, Clefting) syndrome: heterozygous mutation in the p63 gene (R279H) and DNA-based prenatal diagnosis. AB - BACKGROUND: Germline mis-sense mutations in the DNA-binding domain of the p63 gene have recently been established as the molecular basis for the autosomal dominant EEC (Ectrodactyly, Ectodermal dysplasia, Clefting) syndrome. OBJECTIVES: To examine genomic DNA from a 36-year-old woman, her 58-year-old father and her 11-year-old son, all with the EEC syndrome, to determine the inherent p63 mutation and, after genetic counselling, to use knowledge of the mutation to undertake a first-trimester DNA-based prenatal diagnosis in a subsequent pregnancy. METHODS: Fetal DNA was extracted from chorionic villi and used to amplify exon 7 of p63 containing the potential mutation. Direct sequencing and restriction endonuclease digestion (loss of AciI site on mutant allele) were used for DNA-based prenatal diagnosis. RESULTS: We identified a heterozygous arginine to histidine p63 mutation, R279H, in all three affected individuals. Prenatal diagnosis demonstrated a homozygous wild-type sequence predicting an unaffected child: a healthy boy was subsequently born at full-term. CONCLUSIONS: These data expand the p63 gene mutation database and provide the first example of a DNA based prenatal test in this ectodermal dysplasia syndrome. PMID- 11903232 TI - Melasma: histopathological characteristics in 56 Korean patients. AB - BACKGROUND: Melasma is a common acquired symmetrical hypermelanosis characterized by irregular light to dark brown macules and patches on sun-exposed areas of the skin. Its histopathological characteristics are not fully understood. OBJECTIVES: To characterize the histopathological features of facial melasma skin in comparison with adjacent normal skin. METHODS: Biopsies were taken from both melasma lesional skin and adjacent perilesional normal skin in 56 Korean women with melasma. The sections were stained using haematoxylin and eosin, Fontana Masson, diastase-resistant periodic acid-Schiff, Masson trichrome and Verhoeff van Gieson stains, and immunostaining for melanocytes. Data on the changes in number of melanocytes and melanin contents of the epidermis were analysed by a computer-assisted image analysis program. The ultrastructure of the skin was also examined. RESULTS: The amount of melanin was significantly increased in all epidermal layers in melasma skin. The staining intensity and number of epidermal melanocytes increased in melasma lesions. Lesional skin showed more prominent solar elastosis compared with normal skin. Melanosomes increased in number and were more widely dispersed in the keratinocytes of the lesional skin. Lesional melanocytes had many more mitochondria, Golgi apparatus, rough endoplasmic reticulum and ribosomes in their cytoplasm. A dihydroxyphenylalanine reaction was apparent in the cisternae and vesicles of the trans-Golgi network in melanocytes from lesional skin. CONCLUSIONS: Melasma is characterized by epidermal hyperpigmentation, possibly caused both by an increased number of melanocytes and by an increased activity of melanogenic enzymes overlying dermal changes caused by solar radiation. PMID- 11903231 TI - Oral administration of persimmon leaf extract ameliorates skin symptoms and transepidermal water loss in atopic dermatitis model mice, NC/Nga. AB - BACKGROUND: We have previously shown that persimmon leaf extract and its major flavonoid constituent, astragalin, inhibited histamine release by basophils and that oral administration of these substances prior to the onset into an atopic dermatitis (AD) model mouse, NC/Nga, prevented development of dermatitis. OBJECTIVES: This study was designed to assess the clinical therapeutic effect of persimmon leaf extract and astragalin in NC/Nga mice suffering from dermatitis and the dose-response preventive effects of persimmon leaf extract on dermatitis and transepidermal water loss (TEWL). METHODS: The efficacy of persimmon leaf extract or astragalin in NC/Nga mice was judged by measurement of skin severity, scratching behaviour, serum IgE levels or TEWL. RESULTS: Oral administration of persimmon leaf extract (250 mg kg(-1)) or astragalin (1.5 mg kg-1) for 4 weeks into NC/Nga mice with overt dermatitis resulted in a decrease in the severity of the condition. The preventive effect of persimmon leaf extract on the dermatitis was dose-dependent and continuous intake of persimmon leaf extract significantly decreased its onset and development. In addition, TEWL was also suppressed at a persimmon leaf extract dose of 250 mg kg(-1). No significant adverse reaction by these substances could be observed. CONCLUSIONS: These observations suggest that persimmon leaf extract or the flavonoid astragalin may be alternative substances for the management of AD. PMID- 11903233 TI - Differentiation between malignant and benign follicular adnexal tumours of the skin by DNA image cytometry. AB - BACKGROUND: We have demonstrated in previous studies that DNA image cytometry (DNA ICM) can be helpful in detecting malignancy in sebaceous tumours of the Muir Torre syndrome and sweat gland tumours. However, little is known about DNA ICM in cutaneous adnexal tumours with follicular differentiation. OBJECTIVES: To study a larger series of benign and malignant follicular adnexal tumours with DNA ICM. METHODS: We studied 13 malignant follicular tumours (seven trichilemmal carcinomas, five malignant proliferating pilar tumours, one pilomatrix carcinoma) and 55 benign follicular tumours (four tumours of the follicular infundibulum, seven Winer's pores, eight trichilemmomas, two trichofolliculomas, 16 trichoepitheliomas, 13 pilomatrixomas, five trichoblastomas) by DNA ICM. All cases were clear-cut as malignant or benign, respectively, on histopathological criteria. The stemline interpretation according to Bocking et al. (DNA distribution in gastric cancer and dysplasia. In: Precancerous Conditions and Lesions of the Stomach, Zhang YC, Kawai K, eds. Berlin: Springer-Verlag, 1993: 103-20) was performed in all cases. In addition, 5[c]-exceeding events (5cEE) and the 2[c] deviation index (2cDI) were calculated, except in one histopathologically benign tumour, which revealed euploid polyploidization, as the analysis of 5cEE and 2cDI is not valid in that case. RESULTS: A 2cDI threshold of 0.24 proved to be the most reliable marker for the distinction between malignant and benign follicular tumours. On the basis of this feature, all malignant and benign tumours were correctly classified. A specificity of 100% was achieved by all three interpretation methods, but the sensitivity of 2cDI for the detection of malignant tumours was superior to the analysis of 5cEE (sensitivity 77%) and to the stemline interpretation (sensitivity 23%). CONCLUSIONS: DNA ICM may be helpful in distinguishing between malignant and benign follicular tumours. PMID- 11903234 TI - Expression of tyrosinase, MIA and MART-1 in sentinel lymph nodes of patients with malignant melanoma. AB - BACKGROUND: Regional lymph node status is an important predictor of survival in patients with malignant melanoma. Mapping of sentinel lymph nodes using sensitive molecular techniques has recently been introduced. Malignant melanoma is heterogeneous in terms of its biological, immunological and metastatic properties, and melanoma cells exhibit a polymorphous expression of tumour markers. Thus, assays that include multiple markers appear to be more sensitive than single-marker assays. OBJECTIVES: To characterize the molecular profiles of melanoma cells in sentinel lymph nodes employing the mRNA expression of tyrosinase, MIA and MART-1 as markers. METHODS: Samples of sentinel lymph nodes from 17 melanoma patients and 18 control nodes from non-melanoma patients were assayed by reverse transcriptase-polymerase chain reaction, using specific primers for each marker. RESULTS: We found that both tyrosinase and MIA expression were sensitive indicators of micrometastases in sentinel lymph nodes that were negative on routine histopathological examination, and that the finding of micrometastases expressing MART-1 in sentinel lymph nodes was negatively correlated with overall survival. CONCLUSIONS: Characterization of the molecular profiles of melanoma cells constitutes a valid means of detecting metastatic melanoma cells in sentinel lymph nodes, and of predicting the survival of melanoma patients. PMID- 11903235 TI - Long-term results in patients with onychomycosis treated with terbinafine or itraconazole. AB - BACKGROUND: Previously, a double-blind, randomized, multicentre study (LION study) compared the efficacy of continuous terbinafine 250 mg daily for 3 or 4 months with itraconazole pulse therapy 400 mg daily for 3 or 4 months. At the end of the study at week 72 terbinafine proved to be more effective. OBJECTIVES: To perform a 4-year follow-up of the Finnish participants in this study. METHODS: Patients were re-examined clinically and mycologically. RESULTS: Complete clinical and mycological cure with terbinafine for 4 months was 78% compared with 35% with terbinafine for 3 months, 24% with itraconazole for 4 months and 28% with itraconazole for 3 months. CONCLUSIONS: These results suggest that the initial treatment for onychomycosis should be a 4-month continuous course of terbinafine. PMID- 11903237 TI - Melanoma detection rate and concordance between self-skin examination and clinical evaluation in patients attending a pigmented lesion clinic in Italy. AB - BACKGROUND: The early diagnosis of melanoma is based on the collaboration between dermatologists and family doctors, who filter subjects to be referred to a pigmented lesion clinic (PLC). Following growing media coverage, there is increasing concern in the general population about the risk of the 'changing mole', resulting in a progressively increased workload in PLCs. AIM AND METHODS: We investigated the causes of referral to a PLC in a series of 193 attendees seen consecutively at the PLC of the University of Florence. Because the number of naevi is the major risk factor for melanoma in Mediterranean populations, the concordance between self-counting of naevi and the clinical evaluation of a PLC dermatologist in order to classify high-risk individuals was also investigated. RESULTS: Detection of a clinically suspicious lesion at dermatological examination occurred in 13 of 193 subjects referred by general practitioners (6.7%), with three melanomas confirmed histologically (overall detection rate: three of 193, 1.6%). The positive predictive value of the 'presence of a suspicious lesion', the cause of referral in 39.9% of subjects, was 9.1% when based on the gold standard criterion represented by the clinical detection of a suspicious lesion by the dermatologist and 3.8% based on the histological diagnosis of melanoma; the negative predictive value was 94.8% (100% when based on the histological diagnosis of melanoma), suggesting that the clinical detection of a suspicious lesion in subjects with different causes of referral (such as risk factors for melanoma, or the need to be reassured about moles) is unlikely. There was poor agreement between self-evaluation based on the presence of multiple naevi and the dermatological examination (gold standard) for both common and atypical naevi. The highest concordance (kappa = 0.32, 95% confidence interval 0.20-0.43) was associated with a dichotomized count of naevi as up to 50 or more than 50 naevi. CONCLUSIONS: In order to reduce the PLC workload, the filtering role of the family doctor needs to be improved, so that only subjects with a specific suspicious lesion are referred to the PLC. The self-assessment of melanoma risk based on the presence of multiple naevi was not reliable. PMID- 11903236 TI - Terbinafine (Lamisil) treatment of toenail onychomycosis in patients with insulin dependent and non-insulin-dependent diabetes mellitus: a multicentre trial. AB - BACKGROUND: Diabetes mellitus (DM) affects an estimated 175 million people world wide. Approximately one-third of patients with DM have toenail onychomycosis. OBJECTIVES: To determine the efficacy and safety of terbinafine treatment of toenail onychomycosis in patients with DM receiving insulin and/or oral antidiabetic agents. Special interest was focused on potential drug interactions with oral hypoglycaemic substances. METHODS: In a multicentre trial, patients suffering from insulin-dependent DM (IDDM) or non- insulin-dependent DM (NIDDM) with toenail onychomycosis were treated for 12 weeks with oral terbinafine 250 mg daily and followed up to 48 weeks. In addition to clinical, mycological and laboratory investigations, blood glucose levels were monitored. RESULTS: At the end of the trial (week 48), a mycological cure rate of 73% was achieved. The rates of clinical cure and complete cure (mycological cure plus clinical cure) were 57% and 48%, respectively. There was no statistically significant difference between the NIDDM and IDDM groups with respect to the cure rates (P > 0.05). No hypoglycaemic episode was reported and none of the patients had hypoglycaemia during the treatment phase. CONCLUSIONS: With excellent cure rates and a good tolerability profile, terbinafine should continue to be a drug of choice for the treatment of toenail onychomycosis in the rising number of NIDDM patients receiving multiple medication. PMID- 11903238 TI - The clinical spectrum of dystrophic epidermolysis bullosa. AB - BACKGROUND: Dystrophic epidermolysis bullosa (DEB) is a genodermatosis resulting from mutations in COL7A1, the gene encoding type VII collagen. The site and specific nature of the underlying mutation determine the clinical phenotype, which ranges widely from a severe mutilating condition to a relatively mild disorder. OBJECTIVES: To document the clinical spectrum of DEB within a defined complete population. METHODS: Since 1992, when compilation of the U.K. epidermolysis bullosa register began, an exhaustive search for DEB sufferers within the Scottish population has been undertaken and their clinical features comprehensively recorded. RESULTS: One hundred and twenty-eight DEB sufferers have been identified within the Scottish population. In descending order, the frequencies of the different forms of DEB were dominant DEB (DDEB) in 88 individuals (68%), DEB of uncertain inheritance in 24 (19%) and recessive DEB (RDEB) in 16 patients (13%). Within this latter group, nine (7%) had the mutilating Hallopeau-Siemens subtype (RDEB-HS), five (4%) had localized (RDEB loc) and two (2%) had a predominantly flexural (inverse) form of RDEB. During the study, two patients with RDEB died from squamous cell carcinomas (SCCs), one originating in the skin and the second arising in the oesophagus. Gastrointestinal problems such as dysphagia, constipation and anal fissures, and restriction of mouth opening were experienced by the majority of patients with RDEB and by a significant minority of DDEB sufferers. Pseudosyndactyly was most severe in RDEB-HS, all those over 9 years of age having mitten deformities of the hands. Milder pseudosyndactyly or flexion contractures of the fingers were present in younger patients with this subtype, in most adults suffering from other subtypes of RDEB and in 6% of those with DDEB. External ear involvement, a feature not often reported in DEB, was common in RDEB and also occurred in a minority of those with DDEB. Pruriginous lesions and albopapuloid lesions were each present in both DDEB and RDEB. CONCLUSIONS: Most patients with DEB have relatively mild dominantly inherited disease, only a minority suffering from severe recessive subtypes. Scarring, gastrointestinal involvement, albopapuloid lesions and a pruriginosa-like pattern each occur in both DDEB and RDEB. With increasing age, SCC is a major cause of morbidity and mortality. PMID- 11903239 TI - Results of photopatch testing in Rotterdam during a 10-year period. AB - BACKGROUND: Photoallergic contact dermatitis is of importance in a proportion of photodermatoses and can be evaluated through photopatch testing. OBJECTIVES: To conduct a retrospective evaluation of photopatch tests performed in patients with suspected photodermatoses at the clinic at the University Hospital Rotterdam during a 10-year period. METHODS: During the first 5(1/2) years 44 patients were tested with a standard set of 14 allergens, and during the next 4(1/2) years 55 patients were tested with a standard set of 23 allergens. RESULTS: Photocontact reactions were found in 9% and 27% of patients in the two periods, respectively. In the second period, positive reactions were mostly produced by sunscreens. The difference in the percentage of positive photopatch tests was probably caused by the difference in composition of the standard set of allergens (more sunscreens in the second period), this being the only alteration in the test procedure. CONCLUSIONS: The standard set of photoallergens has to be updated periodically. Standardization of the test procedure is needed to compare the test results of different institutions. PMID- 11903240 TI - Do we alter ultraviolet sensitivity in vivo with stratum corneum rehydration? A pilot study and review of the literature. AB - BACKGROUND: Numerous therapeutic schemes recommend topical administration of emollients immediately prior to ultraviolet (UV) B therapy. The rationale behind the clinical improvement is a presumed enhancement of UV transmission through the epidermis. Originating from this clinical observation, there has been some concern as to whether a well-hydrated skin in general might be more susceptible to actinic damage. OBJECTIVES: To investigate whether rehydration of healthy skin causes an altered UVB sensitivity in vivo. METHODS: We determined minimal erythema doses (MEDs) and erythema sum scores (ESSs) after differential rehydration of the skin in 10 healthy volunteers. In each subject six UVB phototests were performed after pretreatment with five different emulsifying ointments (unguentum emulsificans and dilutions with 30, 50, 70 and 90% aqua purificans) plus a negative control. In vivo evaluation of stratum corneum hydration was performed by measurement of electrical capacitance. RESULTS: The results of this randomized, double-blind in vivo study indicated that rehydration of normal stratum corneum with the emulsifying ointments tested did not result in a significantly altered sensitivity to the erythematous effects of UVB irradiation (no significant differences in MED and ESS). Furthermore, there was no correlation between measured stratum corneum hydration and the erythema response of healthy skin. CONCLUSIONS: Although many schemes recommend the administration of emollients prior to UV therapy, there have also been calls for caution, as an uncritical application may interfere with such treatment. We showed that the emulsifying ointments tested exhibited no photoprotective potential and thus are suitable for the pretreatment of psoriasis prior to phototherapy. It has long been discussed whether the effects of emollient pretreatment on response to UV occur only in psoriatic skin or also in healthy skin. Our results indicated that stratum corneum rehydration did not result in a significantly increased erythema response of healthy skin to UVB exposure. With regard to the use of rehydrating cosmetics in everyday life, the outcome of our pilot study is reassuring, as we could not confirm with our experimental design that well-hydrated healthy skin is more prone to actinic damage. PMID- 11903241 TI - Flashlamp pulsed dye laser and argon-pumped dye laser in the treatment of port wine stains: a clinical and histological comparison. AB - BACKGROUND: Port-wine stains (PWS) are congenital vascular malformations occurring in 0.3% of children. The pulsed dye laser is a well established treatment for PWS. OBJECTIVES: To compare, clinically and histologically, the effects of the flashlamp pulsed dye laser with the argon-pumped dye laser in the treatment of PWS. METHODS: Thirty patients were treated on two to four test areas with both laser types using different energy fluences. A flashlamp pulsed dye laser with 0.45 ms pulse duration and a spot size of 5 mm was compared with an argon-pumped dye laser, with a spot size of 1 mm delivered with a robotic scanning laser handpiece (Hexascan) and 70-190 ms pulse duration. Both were tuned to 585 nm. Twelve weeks later the degree of lightening was evaluated and biopsies were taken. To count the vessels the skin sections were stained with CD34 using an immunohistochemical technique. The vessels were divided into three groups by diameter (d): d < 10 microm, 10 < or = d < 20 microm, d > or = 20 microm. RESULTS: The clinical results showed a significantly better lightening using the flashlamp pulsed dye laser than with the argon-pumped dye laser. The histological results showed a significant decrease in the number of vessels of diameter larger than 20 microm in treated compared with untreated lesions. We found no histological difference in the number of vessels between the two laser treatments. However, there was a tendency towards more small vessels (diameter < 10 microm) after one treatment with the flashlamp pulsed dye laser compared with untreated PWS. CONCLUSIONS: The flashlamp pulsed dye laser is clinically superior to the argon-pumped dye laser in the treatment of PWS. PMID- 11903242 TI - Mohs' micrographic surgery using frozen sections alone may be unsuitable for detecting single atypical melanocytes at the margins of melanoma in situ. AB - BACKGROUND: It remains questionable whether micrographic surgery with frozen sections is an appropriate technique for excision of melanoma in situ (MIS) of the lentigo maligna type. Advocates of the technique have interpreted MIS as being histologically defined by nests and contiguous atypical melanocytes on the basal layer. Others, however, have viewed the periphery of MIS as consisting of scattered single atypical melanocytes, a finding that may be difficult or impossible to establish on frozen sections. OBJECTIVES: To examine the reliability of micrographic surgery using frozen sections interpreted by an experienced Mohs' surgeon, in the excision of MIS. METHODS: From a total of 154 specimens, frozen sections from the 50 specimens with margins that were considered difficult to interpret were thawed, sent for routine processing and then examined 'blind' by a dermatopathologist. RESULTS: Using the dermatopathologist's report on paraffin-embedded sections as a reference point, the sensitivity and specificity of frozen sections were calculated to be 59% and 81%, respectively. CONCLUSIONS: Using these histological criteria, micrographic surgery with frozen sections alone is unreliable in the excision of MIS. PMID- 11903243 TI - Teaching dermatology to medical students: a survey of current practice in the U.K. AB - BACKGROUND: In 1993, the General Medical Council recommended that all medical schools should revise their curricula for undergraduate medical education and foster more interdisciplinary collaboration in teaching. In accordance with these recommendations, new curricula have been introduced in U.K. medical schools. OBJECTIVES: To assess the impact of changes in medical curricula on the teaching of dermatology to medical undergraduates. METHODS: A questionnaire was sent to the dermatologists responsible for organizing the teaching of undergraduate dermatology in each of the 24 medical schools in England, Scotland, Wales and Northern Ireland. RESULTS: Replies were received from all schools. Nineteen of the 24 schools had already introduced integrated curricula and the others were changing more slowly. Some dermatology was included in the core curriculum in all schools. Dermatologists in 14 schools contributed to the teaching of basic science and students in 18 schools were able in years 1 and 2 to see patients in primary care (14) and/or the hospital (13). In nine of these schools, students could meet dermatology patients in these early clinical sessions. Nine schools used some problem-based learning (PBL) in addition to other teaching methods, but PBL predominated in four schools and in two of these schools most students never met a dermatologist. Dermatology was a compulsory clinical attachment in 21 schools, but the length of attachments varied and was less than 5 days in four schools. Students had to pass a dermatology assessment at the end of the clinical attachment in 14 schools and there was assessment of knowledge of dermatology in final examinations in all schools. Students had an early opportunity to explore a dermatology topic in depth in 17 schools, and 20 schools offered or were planning to introduce special study modules in dermatology. Interdisciplinary teaching links were common. Resources for out-patient teaching were inadequate in 16 schools and university support poor in 10 schools. Few departments had direct access to the considerable health service funding that is paid to National Health Service Trusts to reimburse the costs of teaching medical students. CONCLUSIONS: In general, dermatology has maintained a reasonable profile in the new undergraduate curricula, but dermatology experience is inadequate in four schools. Dermatologists should maximize opportunities for introducing dermatology into the curriculum by familiarizing themselves with the forces that are driving curriculum reform, participating in curriculum development, keeping abreast of changes in medical education and using opportunities for interdisciplinary teaching. PMID- 11903245 TI - Lupus anticoagulant and venous leg ulceration. AB - BACKGROUND: Most leg ulcers occur in patients with venous insufficiency. However, not all patients with venous insufficiency develop leg ulcers. Recent studies have found that factors causing clotting abnormalities, e.g. anticardiolipin antibody (ACA), are associated with leg ulcers. Although lupus anticoagulant, like ACA, belongs to the group of antiphospholipid antibodies, its presence in patients with venous leg ulceration has not been previously reported. OBJECTIVES: To determine the presence of lupus anticoagulant in patients with venous leg ulceration. METHODS: We investigated the presence of lupus anticoagulant in 27 patients with venous leg ulcers and compared these data with controls. Lupus anticoagulant was evaluated in all subjects by the Russell's viper venom test. RESULTS: Of 27 patients with venous leg ulceration, 16 (59%) were shown to have lupus anticoagulant, while only one of 32 controls (3%) was found to have lupus anticoagulant. Thus, lupus anticoagulant was significantly more frequent in patients with venous leg ulcers than in controls (P < 0.001). CONCLUSIONS: We suggest that lupus anticoagulant could be a hitherto unknown factor contributing to the development of venous leg ulcers. PMID- 11903244 TI - The importance of a dedicated patch test clinic. AB - BACKGROUND: A specialist patch test clinic was set up in April 1997 at the Department of Dermatology, South Infirmary-Victoria Hospital, Cork, Ireland. The number of batteries available was expanded from six to 21 and the routine testing of patients to their own products was introduced, as was prick testing for latex hypersensitivity. OBJECTIVES: To assess the impact of introducing this clinic on the detection of allergic contact dermatitis. METHODS: Patch test results for the first full year of operation of the clinic (1998) were compared with those in the year prior to setting it up (1996). RESULTS: Although the number of patients tested rose after the introduction of the new clinic, the difference was not significant as the number of new dermatology general referrals had also risen. Thirty-one of the 91 patients tested in 1996 had positive patch tests compared with 84 of 158 tested in 1998 (P = 0.0036). Eighteen allergens were detected in 1996 and 53 in 1998. Two patients were positive to their own products in 1996, compared with 12 in 1998 (P = 0.04). The commercial batteries were negative in four of these cases. Three cases of latex hypersensitivity were detected in 1998. CONCLUSIONS: The introduction of a specialist patch test clinic resulted in an increase in detected cases of allergic contact dermatitis. The larger range of batteries available and the more widespread testing of patients' own products were the principal factors involved. PMID- 11903246 TI - Subcutaneous infection by Microsporum gypseum. AB - We report a case of subcutaneous infection caused by the dermatophyte Microsporum gypseum in an immunocompetent host. The patient acquired the infection in the knee through a traumatic inoculation of a splinter. To our knowledge this is the first case of subcutaneous infection caused by this fungus. PMID- 11903247 TI - Cutaneous larva migrans with folliculitis: report of seven cases and review of the literature. AB - Seven patients (four men, three women, mean age 31 years), all returning from the tropics, presented with pruritic folliculitis and creeping eruption. The folliculitis consisted of 20-100 follicular papules and pustules confined to a particular area of the body, mainly the buttocks. The creeping eruption consisted of two to 10 serpiginous or linear burrows 1-5 cm long located either in the same area or in a different area from the folliculitis. Five patients were cured with one to three courses of ivermectin (one course in two cases, two courses in two cases and three courses in one case) and two patients were cured with a 3-day regimen of albendazole. Folliculitis should be added to the dermatological manifestations of cutaneous larva migrans. Treatment is more difficult than in classical forms of cutaneous larva migrans. PMID- 11903248 TI - Calcipotriol for erythema annulare centrifugum. AB - : Erythema annulare centrifugum (EAC) is an uncommon inflammatory skin disease of unknown aetiology. No therapy is currently available. We describe a 73-year-old woman with a 3-year history of EAC that was resistant to topical and systemic glucocorticoids, antifungals, and psoralen plus ultraviolet A treatment. After 3 months of treatment with topical calcipotriol the lesions cleared completely and did not recur during a 6-month follow-up period. Vitamin D analogues may be of value in the therapy of EAC. PMID- 11903249 TI - Cutaneous sarcoidosis during interferon alfa and ribavirin treatment of hepatitis C virus infection: two cases. AB - Interferon-induced sarcoidosis is well documented. We report two new cases of sarcoidosis in two patients with hepatitis C virus infection treated with interferon alfa and ribavirin. These patients developed cutaneous sarcoidosis about 3 months after the beginning of the combination therapy. Spontaneous regression of the lesions was noted after discontinuation of the treatment. There have been more than 20 observations of the appearance or aggravation of this granulomatosis with interferon alfa and more recently with the combination of interferon alfa plus ribavirin. Dermatological signs are found in 50% of cases, and are often diagnostic. Other clinical symptoms of sarcoidosis resemble side effects of interferon. The evolution is fairly stereotypical and is marked by a regression of the lesions following a dose reduction or curtailment of interferon. Interferon alfa acts by stimulating the T-helper (Th) 1 immune response. In addition to its antiviral action, ribavirin also enhances the Th1 response. Indeed, the superiority of the combination of interferon alfa and ribavirin in terms of antiviral action is corroborated by the enhancement of a Th1-type immune reaction by this combination. At the same time, this immune cell reaction triggers a greater granulomatous reaction. PMID- 11903250 TI - Hair darkening in porphyria cutanea tarda. AB - Repigmentation of grey hair is rare, but has been described in several clinical settings. It has most often been reported as a postinflammatory effect, but several drugs, chronic arsenic exposure and coeliac disease have also been cited in addition to darkening as a spontaneous phenomenon. We report two patients with sustained repigmentation of the hair in association with porphyria cutanea tarda. The mechanism for this repigmentation remains elusive, but presumably involves recruitment of outer root sheath melanocytes, which are then activated to form functional hair bulb melanocytes. PMID- 11903251 TI - Regression of multiple viral warts in a human immunodeficiency virus-infected patient treated by triple antiretroviral therapy. PMID- 11903252 TI - Are we going OTT about ITT? PMID- 11903254 TI - Extragenital lichen sclerosus successfully treated with topical calcipotriol: evaluation by in vivo confocal laser scanning microscopy. PMID- 11903255 TI - Dermatological complications of etanercept therapy for rheumatoid arthritis. PMID- 11903256 TI - Treatment of severe psoriasis and psoriatic arthritis with leflunomide. PMID- 11903257 TI - Treatment of erythromelalgia with a serotonin/noradrenaline reuptake inhibitor. PMID- 11903258 TI - Lack of modification of virological status after chemotherapy or radiotherapy for classic Kaposi's sarcoma. PMID- 11903259 TI - Porokeratosis of Mibelli: successful treatment with 5% imiquimod cream. PMID- 11903260 TI - Contact anaphylaxis from natural rubber latex used as an adhesive for hair extensions. PMID- 11903261 TI - Paraneoplastic pemphigus triggered by Castleman's disease. PMID- 11903262 TI - Liquid nitrogen cryotherapy of common warts: cryo-spray vs. cotton wool bud. PMID- 11903264 TI - Topical tacrolimus and pimecrolimus are not associated with skin atrophy. PMID- 11903266 TI - Recurrent angio-oedema and solitary molluscum contagiosum as presenting signs of non-Hodgkin's B-cell lymphoma. PMID- 11903268 TI - AHRQ releases updated health insurance coverage statistics. PMID- 11903269 TI - Tobacco use: all things must pass. PMID- 11903270 TI - A young woman with recurrence of breast cancer. PMID- 11903271 TI - Exercise discussions during cancer treatment consultations. AB - PURPOSE: Physical exercise has been shown to help cancer survivors cope with treatment side effects. The purpose of this study was to examine the role of the oncologist in promoting exercise in cancer survivors. DESCRIPTION OF STUDY: Cancer survivors who had recently completed treatment (N = 311) were mailed a self-administered questionnaire that assessed the following: whether exercise was discussed during any of their treatment consultations; whether they preferred that exercise be discussed during this time; their perceived oncologist approval for exercise; and the amount of exercise they performed during treatment. RESULTS: Descriptive statistics showed that 28.4% of cancer survivors reported that their oncologist initiated a discussion of exercise during their treatment consultation, that 13.9% said that they initiated a discussion, and that 57.8% said that exercise was not discussed. Survivors younger than 60 years of age were more likely to initiate a discussion of exercise than were older survivors. Survivors whose oncologist initiated a discussion of exercise also reported a stronger normative belief for exercise, performing more frequent exercise during treatment, and performing more total minutes of exercise during treatment. Most (82.2%) survivors preferred that the oncologist initiate the discussion of exercise. CLINICAL IMPLICATIONS: These findings indicate that most cancer survivors responding to this survey preferred that their oncologist initiate a discussion of exercise. Such a discussion appears to increase survivor exercise levels during treatment. If confirmed in a prospective, randomized, controlled trial, the implication is that an oncologist-initiated discussion of exercise during treatment consultations may be a cost-effective strategy for promoting exercise in cancer survivors. PMID- 11903272 TI - Lake Superior Rural Cancer Care Project, Part III: provider practice. AB - PURPOSE: Effective methods that encourage rural primary-care physicians to adopt state-of-the-art cancer-management practices are needed. The purpose of this study was to evaluate educational and systems strategies to improve rural primary care physicians' cancer practice behaviors. DESCRIPTION OF STUDY: The Lake Superior Rural Cancer Care Project was a group-randomized, controlled trial conducted with 18 rural communities in the North Central United States over 4 years. Although the unit of analysis was the community, the subjects were 104 primary-care physicians and 2089 rural patients with cancer. The intervention was educational and comprised systems strategies that targeted rural primary-care physicians and their healthcare delivery systems. The outcome measures reported here were physician practice behaviors regarding cancer diagnosis, staging, treatment, clinical trial participation, and post-treatment surveillance. RESULTS: The intervention significantly improved 5 of the 37 cancer practice end points. The overall result of the study did not support the majority of the study hypotheses. Because 16 practice end points were found to be at acceptable performance levels, the possibility of a measurable intervention effect was limited. CLINICAL IMPLICATIONS: Earlier, the authors reported the results of the intervention on providers' cancer management knowledge, which showed significant improvement. The present study findings demonstrated that improving provider knowledge does not necessarily improve practice performance. Changing practice behaviors requires much more effort. Furthermore, interventions found to be effective in other diseases, types of providers, or settings may not work on rural providers for cancer management. PMID- 11903273 TI - Age at diagnosis and quality of life in breast cancer survivors. AB - PURPOSE: The purpose of the study was to determine the relationship between life stage variables (ie, age at diagnosis and years of survival) and quality-of-life (QOL) outcomes in long-term survivors of breast cancer. DESCRIPTION OF STUDY: In this cross-sectional study, 105 long-term survivors of breast cancer participated in a mailed survey assessing QOL. Participants were selected from a cancer center tumor registry using a stratified random-sampling procedure that was based on age at diagnosis. The Quality of Life-Cancer Survivors scale was used to assess QOL outcomes in the physical, psychological, social, and spiritual domains. RESULTS: Long-term survivors of breast cancer who had received diagnoses at an older age (> 65 years) showed significantly (P <.05) worse QOL outcomes in the physical domain, while those who had received diagnoses at a younger age (27-44 years) showed worse QOL outcomes in the social domain than other age groups. A nonlinear relationship was observed, with long-term survivors who had received diagnoses in middle age (45-65 years) showing better QOL outcomes in the physical domain and in overall QOL. Age at diagnosis and years of survival were significant predictors of QOL outcomes. CLINICAL IMPLICATIONS: These findings indicate that the life stage at diagnosis can help to predict long-term QOL outcomes in breast cancer survivors. Educational strategies to help oncology professionals develop a better understanding of the impact of age at diagnosis may be important in developing tailored interventions that respond to the specific needs of breast cancer survivors at each life stage. PMID- 11903274 TI - Reflections on nutritional issues associated with cancer therapy. AB - PURPOSE: It is now recognized that dietitians have a significant role to play in the care of oncology patients, many of whom have problems with nutrition. However, it is increasingly understood that there are important obstacles to assisting patients with nutritional problems in the oncology setting. The purpose of this article is to discuss the incidental findings of two recent studies, highlighting the importance of and problems associated with nutritional issues in patients with a hematologic malignancy. OVERVIEW: This article presents findings from two recent studies with a subset of patients who have hematologic malignancies. This diagnostic subgroup has specific challenges associated with nutrition and eating because of the intensive and aggressive treatments that patients endure. An explorative, descriptive, iterative, qualitative methodology was used for both studies. The focus was on exploring the lived experience of treatment in oncology/hematology for patients (prospective) and caregivers (retrospective). CLINICAL IMPLICATIONS: The findings indicate that the issues surrounding food and eating are considered to be of great significance both to patients who are undergoing intensive treatment for a hematologic malignancy and to their caregivers. The significance of food is not seen purely in relation to its nutritional value, but as an important quality-of-life issue. Eating problems during intensive chemotherapy are perceived as highly stressful and can be, in the case of treatment-related anorexia, life threatening. Recommendations are made for interventions that could reduce the stress and nutritional difficulties for both patients and caregivers. PMID- 11903275 TI - Genetic testing: issues related to privacy, employment, and health insurance. PMID- 11903276 TI - Resources for facilitating back to school programs. PMID- 11903277 TI - Tretinoin for the treatment of acute promyelocytic leukemia. PMID- 11903278 TI - Hemicrania continua: diagnostic criteria and nosologic status. PMID- 11903279 TI - Long-term follow-up of patients treated for chronic headache with analgesic overuse. PMID- 11903280 TI - Hemicrania continua: diagnostic criteria and nosologic status. AB - Proposals for the diagnostic criteria for hemicrania continua (HC) and also for the nosological status of HC are set forth. The clinical constellation of symptoms and signs making up HC consists of: unilaterality without side shift; absolute indomethacin effect; and long-lasting repetitive attacks of varying duration, eventually with a chronic pattern, the pain being mild to severe. For the typical clinical picture of HC, including a positive 'indotest', we propose the term hemicrania continua vera. More or less analogous, but 'indotest negative' clinical pictures have provisionally been termed hemicrania generis incerti (of undetermined nature). At the present level of knowledge, the diagnosis of hemicrania generis incerti should be made mostly by exclusion. HC may possibly best be classified along with chronic paroxysmal hemicrania (CPH) as this is the only other headache absolutely responsive to indomethacin. The bond between these two headaches on the one hand and cluster headache on the other should, at most, be a loose one. Interrelationships of these four classifiable headaches are briefly discussed. PMID- 11903281 TI - Long-term follow-up of patients treated for chronic headache with analgesic overuse. AB - The study aim is to describe the long-term clinical outcome of 102 chronic headache patients with analgesic daily use. They were assessed for daily drug intake (DDI), headache index (HI) and quality of life (QoL) and compared with a parallel group of patients with active chronic daily headache but no analgesic overuse. For the primary study group, baseline 1995 DDI was 1.80 +/- 1.87 and did not differ significantly in 1999. Patients who daily continued to use analgesics had a higher 1995 baseline DDI (t = 2.275, P = 0.025), a longer drug abuse history (t = 2.282, P = 0.025) and a higher DDI (t = 4.042, P < 0.001) 4 years later. At 4 years of follow-up, only one-third of patients initially treated for chronic daily headache and analgesic overuse are successful in refraining from chronic overuse. Those subjects appear to have a persistence for combination analgesic agents; however, their QoL is slightly better than that of patients who revert to episodic headache or continue with chronic daily headache but do not overuse analgesic agents. Persistent analgesic overuse seems to be linked to the length of abuse and to the number of drugs ingested. PMID- 11903282 TI - Physical impairments in cervicogenic headache: traumatic vs. nontraumatic onset. AB - In order to quantify the physical impairments associated with different types of headache, 77 subjects belonging to four different groups (postmotor vehicle accident cervicogenic headache subjects, cervicogenic headache subjects nontraumatic, migraine patients and control subjects) were evaluated using the following variables: posture, cervical range of motion, strength of the neck flexors and extensors, endurance of the short neck flexors, manual segmental mobility, proprioception of the neck, and pain (McGill Pain Questionnaire and the skin roll test). The results of this study showed that postmotor vehicle accident cervicogenic patients have significantly limited active cervical range of motion (in flexion/extension and rotations), present decreased strength and endurance of neck flexors and decreased strength of the extensor muscles. Our results suggest that there are enough differences between the postmotor vehicle accident and nontraumatic cervicogenic headache subjects to warrant caution when analysing the data of these two subgroups together, as several studies have done in the past. The onset of headache is therefore an important variable that should be controlled for when attempting to characterize the physical impairments associated with cervicogenic headache. PMID- 11903283 TI - The prevalence of migraine in patients with bipolar and unipolar affective disorders. AB - There is a well-known association between migraine and affective disorders, but the information is sparse concerning the prevalence of migraine in subgroups of the affective disorders. The present study was undertaken to investigate the prevalence of migraine in unipolar depressive, bipolar I and bipolar II disorders. Patients with major affective disorders (n = 62), consecutively admitted to an open psychiatric ward, were examined with a semi-structured interview based on DSM-IV diagnostic criteria, combined with separate criteria for affective temperaments. Diagnosis of unipolar and bipolar I disorders followed the DSM-IV criteria, while bipolar II disorder encompassed patients with either discrete hypomanic episodes or a cyclothymic temperament. Migraine was diagnosed according to IHS-criteria. Symptoms of migraine were found to be common in these patients, both in those with unipolar depression (46% prevalence of migraine) and in those with bipolar disorders (44% prevalence). Among the bipolar patients there was, however, a striking difference between the two diagnostic subgroups, with a prevalence of 77% in the bipolar II group compared with 14% in the bipolar I group (P = 0.001). These results support the contention that bipolar I and II are biologically separate disorders and point to the possibility of using the association of bipolar II disorder with migraine to study both the pathophysiology and the genetics of this affective disorder. PMID- 11903284 TI - Prevalence and clinical features of migraine in a population of visually impaired subjects in Curitiba, Brazil. AB - To investigate the relevance of lacking or diminished visual input on the expression of migraine, we evaluated its prevalence and clinical features in a population of visually impaired subjects. Between September 1999 and April 2000, 203 visually impaired subjects with a headache inventory were surveyed. Those with headache were assessed according to IHS criteria for the presence of migraine. Migraineurs had their symptoms further detailed through an interview and a headache diary. Of the 104 subjects reporting headaches during the last 6 months, 29 had migraine (14.2%). The prevalence of migraine was not influenced by whether the visual impairment was complete or partial. Mean frequency of migraine attacks was 2.7/month. Most subjects (96%) reported severe and/or moderate attacks. Nausea, vomiting, aggravation by activity and phonophobia were reported by 62%, 37.9%, 86.2% and 96.6% of the subjects, respectively. Visual impairment does not seem to influence prevalence of migraine or its clinical features. PMID- 11903285 TI - Dose, efficacy and tolerability of long-term indomethacin treatment of chronic paroxysmal hemicrania and hemicrania continua. AB - Indomethacin has consistently been proven to provide complete and sustained relief of symptoms in hemicrania continua (HC) and chronic paroxysmal hemicrania (CPH), but is not devoid of side-effects. The goal of this retrospective study is to assess the dose and side-effects of prolonged indomethacin treatment of HC and CPH. Twenty-six patients with either HC or CPH were followed during an average of 3.8 years after onset of treatment with indomethacin. Relief of symptoms occurred within 3 days of treatment, with 84 +/- 32 mg/day of indomethacin. With time, 42% of patients experienced a decrease of up to 60% in the dose of indomethacin required to maintain a pain-free state. Six (23%) patients showed adverse events, mostly gastrointestinal and relieved with ranitidine. No major side-effects were observed. These results indicate that prolonged indomethacin treatment of HC or CPH has a good safety and tolerability profile with a reduction of up to 60% in the initial dose. PMID- 11903286 TI - Headache in magical and medical papyri of ancient Egypt. AB - Despite the intensity with which many scholars have studied the evolution of Egyptian medicine, interdisciplinary studies on the history of headache are scarcely extant. Following a short discussion of historiographical issues, the main objective of this paper is to present a comprehensive and detailed overview on this subject. Scattered references to headache are extracted from so-called magical papyri and from medical texts of the New Kingdom. Although little is known about the quality of headache and about accompanying symptoms, four predominant localizations are distinguished. Due to the lack of precise descriptions it is impossible to establish the retrospective diagnosis of migraine. Explanations of the origin of cephalalgia and of the corresponding therapeutic actions differ according to the nature of the source. In magical papyri, headaches are attributed to the action of demons and supernatural forces, whereas medical papyri emphasize the role of head trauma and of 'pain matter' occurring in the body. Treatment could be magical, pharmacological or surgical. Examples of incantations and prescriptions are analysed in detail. PMID- 11903287 TI - Symptomatic trigeminal-autonomic cephalalgia evolving to trigeminal neuralgia: report of a case associated with dual pathology. PMID- 11903288 TI - Rathke's cleft cyst as a secondary cause of headache:a case report. PMID- 11903289 TI - Melkersson-Rosenthal syndrome presenting as a new daily persistent headache: relief with dapsone. PMID- 11903290 TI - Cluster headache without autonomic symptoms? PMID- 11903291 TI - Hypnic headache and travel across time zones: a case report. PMID- 11903292 TI - Evidence-based migraine therapy. PMID- 11903294 TI - Proceedings and abstracts of the 3rd Franco-Australian Meeting on Hypertension. Corsica, France, June 2001. PMID- 11903295 TI - Renal and cardiac sympathetic baroreflexes in hypertensive rabbits. AB - 1. The purpose of the present study was to assess the changes to renal sympathetic nerve activity (RSNA) baroreflexes during the development of hypertension after renal clipping in conscious rabbits. 2. Rabbits were fitted with a clip on the right renal artery or underwent a sham operation under halothane anaesthesia. A recording electrode was implanted on the left renal nerve 1 week before the experiment, 3 or 6 weeks after the initial operation. During the experiment, drug-induced ramp rises and falls in mean arterial pressure (MAP) were used to produce RSNA and heart rate (HR) baroreflex curves. The RSNA for each experiment was calibrated against maximum RSNA evoked by stimulation of baroreceptor-independent trigeminal afferents. 3. Mean arterial pressure was 20 and 36% higher 3 and 6 weeks after clip implantation, respectively. Renal sympathetic nerve activity baroreflex curves were reset rightwards accordingly, but the shape of the RSNA curves was differentially affected. 4. At both hypertensive periods, MAP-HR baroreflex gain was markedly reduced due to a reduction in curvature. The HR baroreflex range was increased. The RSNA baroreflex gain was reduced at 3 weeks, which was due to a 35% lower RSNA baroreflex range, but was similar to sham animals at 6 weeks. 5. The results show that, in established two kidney, one clip hypertension in rabbits, the sympathetic baroreflex is relatively well preserved but sensitivity of cardiac baroreflexes is attenuated. Therefore, the short-term inhibition of RSNA baroreflexes is not related to the level of blood pressure or the development of secondary changes, such as cardiac or vascular hypertrophy, but may be related to circulating angiotensin, which is known to increase at this time. PMID- 11903296 TI - Does it make sense to develop new centrally acting cardiovascular drugs? AB - 1. The autonomic nervous system plays a pivotal role in modulating all the components of the cardiovascular regulation. Therefore, one can assume that drugs targeting this system may be useful in the management of several cardiovascular diseases. 2. Drugs acting on central nervous system centres seem to be modulators rather than blockers; as such, they are expected to preserve the contraregulatory processes and to generate only a few side effects. 3. Because the sympathetic nervous system is largely involved in the regulation of vasomotor tone, centrally acting antihypertensive drugs were developed first. 4. Recently, new leader compounds selective for non- adrenergic imidazoline recepetors have been synthetized. Although such drugs have no capacity to activate alpha2 adrenoceptors, they have been proven to be hypotensive. These drugs are expected to be even better tolerated than the currently available centrally active drugs. They may also have additional beneficial effects. 5. Here, the experimental evidence suggesting that such drugs may be useful in the management of some cardiac arrhythmias and/or left ventricular dysfunction will be reviewed. PMID- 11903297 TI - Paring down on Descartes: a review of brain noradrenaline and sympathetic nervous function. AB - 1. The conceptual framework of mind-body interaction can be traced back to the seminal observations of the French philosopher and mathematician Rene Descartes (1596-1650). Descartes succeeded in eliminating the soul's apparent physiological role and established the brain as the body's control centre. 2. While the pivotal role played by the central nervous system (CNS) in the maintenance of physiological and psychological health has long been recognized, the development of methods designed for the direct examination of human CNS processes has only recently come to fruition. 3. There exists a substantial body of evidence derived from clinical and experimental studies indicating that CNS monoaminergic cell groups, in particular those using noradrenaline as their neurotransmitter, participate in the excitatory regulation of the sympathetic nervous system and the development and maintenance of the hypertensive state. 4. In essential hypertension, particularly in younger patients, there occurs an activation of sympathetic nervous outflows to the kidneys, heart and skeletal muscle. The existence of a correlation between subcortical brain noradrenaline turnover and total body noradrenaline spillover to plasma, resting blood pressure and heart rate provides further support for the observation that elevated subcortical noradrenergic activity subserves a sympathoexcitatory role in the regulation of sympathetic preganglionic neurons of the thorocolumbar cord. PMID- 11903298 TI - Effect of alpha-adrenoceptor blockade on the 0.4 Hz sympathetic rhythm in conscious rats. AB - 1. The present study examined the origin of the 0.4 Hz rhythm in renal sympathetic nerve activity (RSNA) in rats. It was anticipated that, after elimination of 0.4 Hz oscillations of arterial pressure (AP) by alpha adrenoceptor blockade, the persistence or disappearance of a 0.4 Hz rhythm in RSNA would point to an endogenous (central oscillator) or baroreflex origin, respectively. 2. Arterial pressure and RSNA were recorded in seven conscious rats, before and after acute alpha-adrenoceptor blockade with phentolamine (5 mg/kg, i.v.). In each condition, power and coherence spectra were calculated over 15 min periods of rest. 3. In control conditions, highly coherent AP and RSNA oscillations were observed near 0.4 Hz. After phentolamine administration, spectral power in the mid-frequency (0.27-0.74 Hz) band was significantly reduced for both AP and RSNA and maximum power was shifted towards 0.7 Hz. 4. The disappearance of the RSNA rhythm at 0.4 Hz after phentolamine administration favours the hypothesis of a baroreflex origin. The new oscillation near 0.7 Hz can derive either from the activity of a previously unrecognized central oscillator or from a faster feedback mechanism involving cotransmitters of noradrenaline acting with shorter time constants (e.g. ATP). PMID- 11903299 TI - Sympathetic nerve biology in essential hypertension. AB - 1. Although the importance of sympathetic nervous activation in the pathogenesis of essential hypertension is well documented, the exact pathophysiology of the sympathetic nervous dysfunction present remains to be delineated. There are several possible explanations for the increased spillover of noradrenaline from the kidneys and heart to plasma, a key piece of evidence supporting the neurogenic basis of essential hypertension, in addition to the obvious one of an increased rate of sympathetic nerve firing. 2. The possibility that there may be an increase in the density of sympathetic innervation in human hypertension, well documented in the spontaneously hypertensive rat, is currently under investigation by us. 3. Adrenaline cotransmission is present in the cardiac sympathetic nerves of patients with essential hypertension, presumptive evidence of their exposure to high levels of stress and a possible basis for the observed increase in cardiac noradrenaline spillover, through presynaptic augmentation of noradrenaline release. 4. Phenotypic evidence exists also of faulty noradrenaline reuptake into the sympathetic nerves of the heart in essential hypertension, an abnormality that would amplify the sympathetic neural signal by impairing removal of noradrenaline from the synaptic cleft. PMID- 11903300 TI - Neural pathways from the lamina terminalis influencing cardiovascular and body fluid homeostasis. AB - 1. The lamina terminalis, a region of the brain with a high concentration of angiotensin AT1 receptors, consists of three distinct nuclei, the median preoptic nucleus, the subfornical organ and organum vasculosum of the lamina terminalis (OVLT). These latter two regions lack a blood-brain and detect changes in plasma angiotensin (Ang) II concentration and osmolality. 2. Efferent neural pathways from the lamina terminalis to the hypothalamic paraventricular and supraoptic nuclei mediate vasopressin secretion in response to plasma hypertonicity and increased circulating levels of AngII. 3. Studies using the neurotropic virus pseudorabies, which undergoes retrograde transynaptic neuronal transport following injection into peripheral sites, show that neurons in the lamina terminalis have efferent polysynaptic neural connections to the peripheral sympathetic nervous system. Some of these neurons have been shown to have polysynaptic connections to the kidney and to express AT1 receptor mRNA. We propose that circulating AngII acts at AT1 receptors in the subfornical organ and OVLT to influence the sympathetic nervous system. It is likely that the neural pathway subserving this influence involves a synapse in the hypothalamic paraventricular nucleus. 4. The lamina terminalis may exert an inhibitory osmoregulatory influence on renin secretion by the kidney. This osmoregulatory influence may be mediated by inhibition of renal sympathetic nerve activity and appears to involve a central angiotensinergic synapse. 5. The lamina terminalis exerts an osmoregulatory influence on renal sodium excretion that is independent of the renal nerves and is probably hormonally mediated. PMID- 11903301 TI - Glucocorticoid-induced hypertension: from mouse to man. AB - 1. Adrenocorticotrophic hormone (ACTH) raises blood pressure in humans, sheep, rat and mouse. In rat and humans, but not sheep, the hypertension can be explained by glucocorticoid excess. 2. In both rat and humans, the hypertension is associated with a rise in cardiac output and renal vascular resistance. 3. In both rat and humans, the nitric oxide system is implicated in glucocorticoid hypertension. 4. In both rat and humans, hypertension due to naturally occurring glucocorticoids is not prevented by drugs that block classical glucocorticoid or mineralocorticoid receptors. 5. Abnormalities in glucocorticoid metabolism may contribute to some forms of 'essential' hypertension. PMID- 11903302 TI - Coronary dilatation reserve in experimental hypertension and chronic heart failure: effects of blockade of the renin-angiotensin system. AB - 1. The aim of the present study was to investigate left and right ventricular (LV and RV, respectively) coronary vasodilatation reserve (CVR; fluorescent microsphere technique) in rats with hypertension (spontaneously hypertensive rats (SHR)) or congestive heart failure (CHF) and the effects of early and chronic renin-angiotensin system (RAS) blockade thereupon. 2. In adult SHR, both LV and RV CVR were impaired, especially in the non-hypertrophied RV, the main factor involved being coronary vascular remodelling. Blockade of the RAS normalized both LV and RV CVR, mainly through the prevention of hypertension and suppression of the resulting pericoronary fibrosis. 3. In postischaemic CHF rats, there was an early and severe degradation of LV and RV CVR that developed before any significant vascular remodelling and appeared to be linked to the deterioration of cardiac hypertrophy and haemodynamics. This degradation in CVR further worsened over the longer term due to late-developing pericoronary fibrosis and endothelial dysfunction. Blockade of the RAS had no early effects on LV and RV CVR, but improved RV CVR over the long term, mainly by limiting RV hypertrophy and by preventing the development of pericoronary fibrosis and coronary endothelial dysfunction. 4. In kallikrein-kinin system-deficient mice, CVR was not different from that of wild-type mice, suggesting that this system is not implicated in normal CVR regulation. PMID- 11903303 TI - Mineralocorticoids and cardiac fibrosis: the decade in review. AB - 1. Over the past decade, aldosterone has been shown to have direct extra epithelial actions and substantial (patho)physiological roles in the cardiovascular system in the context of inappropriate salt status. In experimental studies on uninephrectomized rats given 0.9% NaCl solution to drink, these include blood pressure elevation via activation of circumventricular mineralocorticoid receptors in the central nervous system and production of pressure-independent cardiac hypertrophy and fibrosis by a direct effect on the heart. 2. In the Randomized Aldactone Evaluation Study (RALES) trial, patients with severe congestive heart failure (CHF) were continued on their current therapy (angiotensin-converting enzyme inhibitor, diuretic etc.) and given either placebo or spironolactone at an average dose of 26 mg/day. Mineralocorticoid receptor inhibition was accompanied by a 30% improvement in mortality and 35% less hospitalization, striking confirmation of a pathophysiological role for aldosterone in CHF. 3. Although the current basic and clinical studies are conflicting, there is evidence both for aldosterone synthesis by the failing human heart and for substantial cardiac metabolism of aldosterone. The extent to which this potential paracrine source for aldosterone may be involved in cardiac hypertrophy and cardiac fibrosis remains to be established. 4. Belatedly, aldosterone-induced proteins (e.g. serum and glucocorticoid-regulated kinase (SGK)) have been identified in epithelial mineralocorticoid target tissue. Studies are currently in progress on the cellular and molecular mechanisms involved in the coronary vasculitis provoked early in the mineralocorticoid/salt model, which, in turn, appears to trigger the subsequent perivascular and interstitial fibrotic response. PMID- 11903304 TI - Environmental and genetic determinants of intima-media thickness of the carotid artery. AB - 1. The aim of the present study was to investigate carotid intima-media thickness (CIMT) in relation to anthropometric, environmental and genetic factors, as well as cholesterol and blood pressure levels. 2. The study sample was composed of 89 families, with no documented cardiovascular disease, consisting of 369 subjects (aged from 10 to 54 years) from the Stanislas cohort. 3. Carotid intima-media thickness was measured by B-mode ultrasonography. Fifteen genetic markers, including genes involved in lipid metabolism, the regulation of blood pressure, thrombosis, platelet function and endothelial cell adhesion, were studied by multiplex assay. 4. The effects of gender, age, smoking, alcohol, body mass index, cholesterol, blood pressure and genetic factors were studied using ANOVA and bivariate and regression analyses. 5. Segregation analysis was also performed to estimate the contribution of genetic and environmental factors to CIMT variability. 6. Carotid intima-media thickness values were not affected by age or by gender up to 18 years of age. Thereafter, CIMT values increased sharply in men and remained significantly higher than in women. 7. Approximately 30% of CIMT variability was attributable to genetic factors. Associations between CIMT and polymorphisms in the apolipoprotein CIII, cholesteryl ester transfer protein, methylene tetrahydrofolate reductase and fibrinogen genes were observed and explained approximately 20% of CIMT variation in men. 8. In women, none of the studied polymorphisms was associated with CIMT variation. 9. Our study gives new perspectives for understanding CIMT variability in healthy middle-aged subjects. PMID- 11903305 TI - Local pulse pressure is a major determinant of large artery remodelling. AB - 1. The aim of the present brief review is to show that the pulsatile component of blood pressure is a stronger determinant of large artery remodelling than the steady component (i.e. mean blood pressure). 2. Pulse pressure, which is a strong determinant of cardiovascular events, including coronary heart disease and stroke, is increased when large arteries stiffen. Local pulse pressure, measured with applanation tonometry in normotensives and patients with essential hypertension, explains a significant part of the variance of intima-media thickness at the site of the carotid artery, a proximal elastic artery, whereas mean blood pressure does not contribute. Local pulse pressure has no influence on intima-media thickness at the site of the radial artery, a distal muscular artery that undergoes very little stroke change in diameter. 3. The decrease in carotid pulse pressure is also a major determinant of the regression of carotid intima media thickness after antihypertensive treatment. Local pulse pressure can influence not only intima-media thickness, but also internal diameter. Indeed, there is a significant association between the lumen enlargement of the ascending aorta in patients with Marfan syndrome and pulse pressure. In addition, carotid pulse pressure is positively correlated with carotid internal diameter in normotensives and hypertensives, and the decrease in carotid internal diameter during long-term antihypertensive treatment is influenced by the decrease in carotid pulse pressure and not by the reduction in mean blood pressure. 4. We suggest that the effects of pulse pressure on large artery remodelling may explain part of its predictive value on cardiovascular events. PMID- 11903307 TI - Alcohol and endothelial function: a brief review. AB - 1. In spite of the dose-related effects of alcohol consumption to increase blood pressure, regular light to moderate alcohol intake appears to confer protection against both coronary artery disease and ischaemic stroke. In contrast, heavy alcohol consumption increases the risk of coronary artery disease and the risk of both haemorrhagic and ischaemic stroke. 2. Effects of alcohol consumption on endothelial cell function may be relevant to these disparate effects on cardiovascular outcomes. In in vitro animal studies, low doses of alcohol have been demonstrated to increase release of nitric oxide and augment endothelium mediated vasodilatation, whereas higher doses impair endothelium-dependent relaxation responses. In contrast, chronic administration of alcohol to rats has generally been associated with tolerance to the acute inhibitory effects of alcohol on endothelium-mediated vasodilatation and may even result in augmentation of such responses. 3. The few human studies to date that have examined the effects of alcohol on endothelial function have focused on postischaemic flow-mediated dilation of the brachial artery (FMD). Although blunted FMD responses have been reported in alcoholic subjects, acute administration of alcohol or short-term interventions to reduce alcohol intake have had no effect to either improve or impair FMD. 4. Further studies in humans assessing acute and longer term dose-related effects of alcohol on endothelial function in both conduit and resistance vessels will be necessary if the relevance of the findings from in vitro and in vivo animal studies are to be understood in the context of the complex interrelationships of alcohol with cardiovascular disease. PMID- 11903306 TI - Analysis of agonist-evoked nitric oxide release from human endothelial cells: role of superoxide anion. AB - 1. Dichlorofluorescein oxidation and electrochemical monitoring of in situ nitric oxide (NO) release from cultured human endothelial cells reveals that agonists such as thrombin and histamine simultaneously stimulate transient superoxide production. 2. The duration of *NO release was increased only in the simultaneous presence of extracellular L-arginine and exogenous superoxide dismutase. In contrast, the inhibition of membrane reduced nicotinamide adenine dinucleotide (phosphate) oxidases, the major source of *O2- in endothelial cells, did not prolong *NO release, although extracellular L-arginine was also present. Comparison of these two experimental conditions suggested that H2O2 was involved in the extension of the *NO signal. 3. The present study demonstrates that, in the absence of external L-arginine, *O2- production does not constitute the major pathway controlling the duration of agonist-induced *NO signal. These results suggest that L-arginine and H2O2 act jointly to maintain nitric oxide synthase in an activated form. PMID- 11903308 TI - Role of arterial smooth muscle tone and geometry in the regulation of peripheral conduit artery mechanics by shear stress. AB - 1. Although arterial blood flow is recognized as an important modulator of vascular tone and geometry, the effect of acute changes in shear-stress on conduit artery mechanics has not been fully investigated in humans because of technical limitations. 2. To assess, respectively, the effects of decreases and increases in flow and shear stress on radial artery tone and mechanics, arterial pressure (photoplethysmography), total blood viscosity, radial artery internal diameter, wall thickness (echotracking) and blood flow (Doppler) were measured in healthy volunteers (mean (+/-SEM) age 25 +/- 1 years) during a distal flow arrest (n=12) and hand skin heating (n=18). 3. Radial artery flow decreased from 31 +/- 4 to 7 +/- 1 10(-3) L/min during distal flow arrest (P < 0.001) and increased from 10 +/- 2 to 22 +/- 4 and 69 +/- 6 10(-3) L/min during heating (P < 0.001). At mean arterial pressure, these changes in flow were respectively associated with a parallel flow-dependent reduction and increase in diameter and midwall stress. There was no significant modification in mean elastic modulus. Compliance did not change when flow decreased and only increased at the highest level of flow. Finally, the cross-sectional compliance and incremental modulus were fitted as functions of midwall stress. The decrease in flow was associated with an upward shift of the modulus-midwall stress curve and a downward shift of the compliance-midwall stress curve. The increase in flow was associated with a downward shift of the modulus-midwall stress curve and an upward shift of the compliance-midwall stress curve at each level of wall shear stress. 4. By using two different procedures, we obtained similar results concerning the direct effects of increases and decreases in flow on stiffness of the arterial wall and on arterial compliance and demonstrated the presence of a flow-dependent regulation of arterial smooth muscle tone of peripheral conduit arteries in humans. PMID- 11903309 TI - Vasculoprotective effects of oestrogens. AB - 1. The rationale for preclinical research on the atheroprotective effect of oestrogens is based on the epidemiological evidence that women are protected against the clinical complications of atherosclerosis until menopause. However, this protection, probably due to sex hormones, is progressively lost within the years following menopause. 2. In addition, numerous studies have clearly demonstrated the atheroprotective effect of oestrogens in all animal models. 3. In the present paper, we first summarize our understanding of the pathophysiology of atherosclerosis. We then focus on the recognized target of oestradiol (E2) in the vessel wall: the classical target, namely the endothelium, and a recently characterized target, namely cells of the inflammatory-immune system. Finally, we discuss how unknown mechanisms in atherosclerosis could be responsible for the absence of effect of hormone-replacement therapy in the Heart and Estrogen/ progestin Replacement Study (HERS). PMID- 11903310 TI - 'Slow pressor' hypertension from low-dose chronic angiotensin II infusion. AB - 1. Angiotensin (Ang) II causes growth-related effects on vascular smooth muscle cells in vitro and in vivo. 2. Chronic infusions of AngII systemically, at doses that are initially subpressor, result in slowly progressive increases in arterial pressure ('slow-pressor' hypertension). It has been suggested that the hypertension is due to induced growth in systemic resistance vessel walls by the AngII infusions. 3. We report the results of several studies investigating whether there are also induced structural changes in renal resistance vessels during chronic AngII infusions. We have developed models in Sprague-Dawley rats in which low-dose AngII infusions, either into the renal artery (thus restricting the effects to the kidney) or systemically, result in hypertension. 5. In both models, we have evidence suggesting that chronic AngII infusions have resulted in apparent structurally induced reductions in renal vasculature lumen upstream to the glomerulus. 6. The role of these renal changes in the development of the hypertension remain to be determined. PMID- 11903311 TI - Large artery stiffness: structural and genetic aspects. AB - 1. Large artery stiffness is a principal determinant of pulse pressure and both are related to cardiovascular mortality independently of other major risk factors. A clearer understanding of the structural and genetic processes that contribute to large artery properties may provide novel approaches to therapy. 2. Age, atherosclerosis and gender are three important factors that contribute to large artery stiffening. Each influences the artery elastic matrix and its relationship to medial smooth muscle cells. Genetic and hormonal modulation of the extracellular matrix proteins and their regulators, including matrix metalloproteinases (MMPs), may account for some interindividual differences. 3. In a study of 213 healthy individuals and 105 patients with coronary artery disease (CAD), we examined whether stromelysin-1 (MMP-3) genotype, determined by the 5A/6A promoter polymorphism, influences large artery stiffening. In healthy individuals, the 5A/5A genotype was linked with stiffer large arteries and higher systolic blood pressure compared with other genotypes. 4. Genetic variation in the extracellular matrix protein fibrillin-1, using a pentanucleotide repeat polymorphism, was assessed as a potential determinant of large artery stiffness in patients with CAD. The 2-3 genotype was associated with stiffer large arteries, higher pulse pressure and more severe CAD than other genotypes. 5. Females experience a greater increase in large artery stiffness with age than males, with a time-course suggestive of sex steroid modulation. The mechanisms mediating such gender differences have not been established, but the known regulatory role of sex steroids with respect to MMPs likely contributes. 6. The demonstration that genetic and hormonal modulation of extracellular matrix components and MMPs contributes to age, atherosclerotic and gender-related differences in large artery mechanical properties suggests these proteins may be important targets for therapy. PMID- 11903312 TI - Renin gene expression: the switch and the fingers. AB - 1. Now that many of the factors and control elements that regulate renin transcription have been identified, the scene is set to address the question of the mode of control. 2. Based on current gene control theories, either renin gene transcription in each cell undergoes gradual responses over a continuous range or transcription is switched completely on or completely off. The latter model of 'binary' or 'variegated' expression fits with observations such as the 'recruitment' of new cells for renin expression during strong physiological stimulation and the progressive switching off of expression during development. 3. The renin gene offers an excellent general model for testing the mode of control of genes that are subject to continuous modulatory influences from the demands of physiological perturbations. This is because the promoter is well characterized and is subject to the influence of a strong far-upstream enhancer, one of the key elements of the variegation model. 4. Renin is also controlled at the post-transcriptional level and this, like transcriptional control, involves cAMP mechanisms. We have cloned the human and mouse homologues of a protein (ZNF265) that is important in renin mRNA processing and stability. This uses 'zinc fingers' to bind the mRNA. The role of this and other proteins in splicing and stabilization of mRNA is now being elucidated. 5. Unravelling the mechanisms that determine rate of supply of renin mRNA to the biosynthetic machinery is being assisted by advances in concepts and techniques in the rapidly moving field of genomics. PMID- 11903313 TI - Phenotypic and genetic heterogeneity of familial hyperkalaemic hypertension (Gordon syndrome). AB - 1. Familial hyperkalaemic hypertension (FHH), also called pseudohypoaldosteronism type II (PHA2) or Gordon syndrome, is a rare Mendelian-form of low-renin hypertension. The first cases of FHH were reported approximately 30 years ago and they described the peculiar biochemical abnormalities (i.e. hyperkalaemia and hyperchloraemic acidosis despite a normal glomerular filtration rate). 2. Since then, more than 90 single cases and families have been reported in the literature. These various reports show marked differences in phenotype. 3. Our group has now collected 14 unrelated pedigrees originating from different parts of France and Europe. We confirm the large variations in the age of discovery and in the severity of the biochemical abnormalities from one individual to another and from one family to another one. 4. Blood pressure levels have no significant relationship with hyperkalaemia or hyperchloraemia, but there is a positive relationship with age, as in the normal population. 5. Analyses of clinical features and Mendelian segregation in our families demonstrate autosomal-dominant inheritance, as expected from the literature. 6. Efforts have been made in the past years to unravel the gene responsible for the disease. Until now, a primary responsibility of the gene encoding the thiazide-sensitive Na-Cl cotransporter (SLC12A3) has been excluded in PHA2 families. Three loci have been identified on chromosomes 1 (PHA2A), 17 (PHA2B) and 12 (PHA2C). 7. More recently, analysis of three additional pedigrees, including 10 affected subjects, with over 25 members allowed us to demonstrate further genetic heterogeneity and the existence of at least a fourth locus. 8. The genetic heterogeneity of this syndrome, and thus the variety of molecular defects, suggests the role of either several new components of the same pathway, multiple aldosterone- regulated effectors or direct or indirect partners of the Na-Cl cotransporter. PMID- 11903314 TI - Sex, genes and blood pressure. AB - 1. Throughout most of life, males have higher average blood pressures than females. This sexual dichotomy may be related to genetic factors including the X and Y sex chromosomes and genes that control sex steroids. Resultant physiological differences between men and women may also be relevant to the quantitative variation of blood pressure within the sexes. 2. The present overview collates our published and novel sex-related genetic data in relation to blood pressure from the Victorian Family Heart Study. These include a multipoint quantitative linkage analysis of the X chromosome and genetic association studies of single nucleotide polymorphisms (SNP) of the Y chromosome and genes encoding the androgen receptor (AR), oestrogen receptor alpha (ERalpha), 5alpha-reductase types I and II (SRD5A1 and SRD5A2) and aromatase (CYP19). 3. Systolic blood pressure (SBP) was linked (Z=3.3, genome-wide P < 0.05) to a region of the X chromosome that encompassed the AR gene and the Y chromosome was associated with diastolic blood pressure (DBP; P=0.03). In new analyses, we observed a possible association between a SNP in AR and DBP in 369 males (84.5 vs 82.1 mmHg for genotype A vs genotype B, respectively; P=0.06) and a significant association between haplotypes of the Y chromosome and AR SNP in males (P=0.01) with a difference of nearly 6 mmHg DBP between extreme groups. Associations were also observed for polymorphisms of SRD5A1 and ERalpha with DBP and SBP in males, respectively. 4. The findings indicate that genes related to sexual phenotypes may be relevant to the normal variation in blood pressure, even within the sexes. Further genetic and physiological analyses will be required to confirm these observations and to determine the mechanisms of action and the nature of any interactions. PMID- 11903315 TI - New elements in human renin promoter involved in cell-specific expression. AB - 1. The renin-angiotensin system plays a major role in blood pressure regulation and electrolyte homeostasis through the action of angiotensin (Ang) II. The first and rate-limiting step in the production of AngII is the conversion of angiotensinogen into AngI, which is catalysed by the aspartyl protease renin (EC 3.4.23.15). Circulating active renin is mainly synthesized, processed and secreted by the juxtaglomerular cells within the kidney. 2. To determine the renin 5'-flanking sequences involved in cell and tissue specificity, ex vivo and in vivo studies were performed. Several constructs of various lengths of renin promoter linked to the luciferase gene were first tested ex vivo by transfection in primary cultures of human chorionic cells. The constructs giving a high and specific expression in renin-producing cells were then tested in vivo in a transgenic mice model. 3. The reporter gene chosen to generate transgenic mice was LacZ and the screening was performed in embryos at the embryonic day (E) 15 stage, at which mouse renin is expressed in the developing vessels of the kidney. 4. Only constructs containing more than 5.7 kb of the human renin promoter lead to specific expression of beta-galactosidase in the kidney. 5. Our results demonstrate that the human renin distal promoter region allows a more restricted expression of LacZ in the renin-expressing cells in transgenic mice. PMID- 11903316 TI - The kallikrein-kinin system in humans. AB - 1. Kinin peptides are implicated in many physiological and pathological processes, including the regulation of blood pressure and sodium homeostasis, inflammation and the cardioprotective effects of preconditioning. In humans, the plasma and tissue kallikrein-kinin systems (KKS) generate bradykinin and kallidin peptides, respectively. 2. We established methodology for the measurement of bradykinin and kallidin peptides and their metabolites in order to study the function of the plasma and tissue KKS in humans. 3. Bradykinin peptides were more abundant than kallidin peptides in blood and cardiac atrial tissue, whereas kallidin peptides were predominant in urine. The levels of kinin peptides in tissue were higher than in blood, confirming the primary tissue localization of the KKS. 4. Angiotensin-converting enzyme inhibition increased blood levels of bradykinin and kallidin peptides. 5. Blood levels of kallidin peptides were suppressed in patients with severe cardiac failure, indicating that the activity of the tissue KKS is suppressed in this condition. 6. Bradykinin peptide levels were increased in the urine of patients with interstitial cystitis, suggesting a role for these peptides in the pathogenesis and/or symptomatology of this condition. 7. Cardiopulmonary bypass, a model of activation of the contact system, activated both the plasma and tissue KKS. 8. Measurement of individual bradykinin and kallidin peptides and their metabolites gives important information about the operation of the plasma and tissue KKS and their role in physiology and disease states. PMID- 11903317 TI - Angiotensin I-converting enzyme and metabolism of the haematological peptide N acetyl-seryl-aspartyl-lysyl-proline. AB - 1. Angiotensin I-converting enzyme (ACE) has two homologous active N- and C terminal domains and displays activity towards a broad range of substrates. The tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) has been shown to be hydrolysed in vitro by ACE and to be a preferential substrate for its N-terminal active site. This peptide reversibly prevents the recruitment of pluripotent haematopoietic stem cells and normal early progenitors into the S-phase. 2. Angiotensin I-converting enzyme inhibitors, given as a single dose to normal subjects or during long-term treatment in hypertensive patients, result in plasma AcSDKP levels five- to six-fold higher and urine concentrations 40-fold higher than those of control subjects and/or patients. Thus, AcSDKP is a natural peptide hydrolysed by the N-terminal domain of ACE in vivo. In addition, ACE may be implicated in the process of haematopoietic stem cell regulation by permanently degrading this natural circulating inhibitor of cell entry into the S-phase. 3. Besides hydrolysis by ACE, the second very effective mechanism by which AcSDKP is cleared from plasma is glomerular filtration. Because of its high sensitivity and specificity, the measurement of AcSDKP in plasma and urine provides a valuable tool in screening specific inhibitors of the N-terminal domain of ACE and in monitoring ACE inhibition during chronic treatment. 4. The long-term consequences of AcSDKP accumulation are not known. During chronic ACE inhibition in rats, AcSDKP levels slightly increase in organs with high ACE content (kidneys, lungs). To significantly increase its concentration in target haematopoietic organs (the extracellular fraction of bone marrow), AcSDKP has to be infused on top of a captopril-based treatment. 5. A selective inhibitor of the N-domain of ACE in vitro and in vivo has been identified recently. The phosphinic peptide RXP 407 does not interfere with blood pressure regulation, but does increase, dose dependently, plasma concentrations of AcSDKP in mice, in contrast with lisinopril, which affects the metabolism of both AcSDKP and angiotensin I. N Terminal-selective ACE inhibitors may be used to selectively control AcSDKP metabolism in target haematopoietic organs. This new therapeutic strategy may be of value for protecting haematopoietic cells from the toxicity of cancer chemotherapy. PMID- 11903318 TI - Interactions between sodium and angiotensin. AB - 1. Increased sodium intake causes decreased formation of angiotensin (Ang) II and increased AngII causes increased Na+ retention. 2. Increased sodium intake and increased AngII causes cardiac hypertrophy, but decreased sodium intake regresses cardiac hypertrophy despite high AngII levels. Likewise, decreased Na+ and blockers of the renin-angiotensin system (RAS) in neonatal rats have similar effects on subsequent blood pressure development. 3. Cardiac hypertrophy due to renal hypertension does not regress when the RAS is blocked and rats are on a high salt intake. Likewise, sodium restriction alone does not cause regression; combination of reduced NaCl intake and RAS blockade is required. 4. High doses of perindopril and losartan in combination cause a syndrome in rats on 0.2% NaCl that leads to profound hypotension, polyuria, renal impairment and involution of the heart and death. This is reversed or prevented by a high (4%) NaCl intake, which also prevents the plasma angiotensinogen depletion that occurs with combined blockade on 0.2% NaCl intake. 5. Intake of NaCl and AngII interact at many levels. It is postulated that there is an important interaction at the cellular level that can explain the above events. PMID- 11903319 TI - Different aortic reflection wave responses following long-term angiotensin converting enzyme inhibition and beta-blocker in essential hypertension. AB - 1. In arterial hypertension, aortic wave reflections contribute to determining central systolic and pulse pressures. The present study assessed the central pressure alterations at the level of the common carotid artery following 1 month treatment with perindopril or atenolol and investigated during the 8 h following drug intake. 2. Twenty patients suffering from permanent hypertension were included after a 4 week run-in placebo period in a double-blind, randomized cross over study comparing the angiotensin-converting enzyme (ACE) inhibitor perindopril with the beta-blocker atenolol during a 4 week treatment period. 3. Before and during the 8 h after drug intake, serial measurements included brachial artery systolic and diastolic blood pressures (SBP and DBP, respectively; mercury sphygmomanometer), carotid artery SBP and pulse pressure (PP; applanation tonometry), aortic pulse wave velocity (Complior; Colson, Les Lilas, France) and arterial wave reflections from the aorta (applanation tonometry; Sphygmocor; PWV Medical, Sydney, NSW, Australia). 4. Both treatments decreased brachial and carotid artery SBP, DBP and PP. Heart rate and pulse wave velocity decreased following atenolol (P < 0.001). Pulse wave velocity was reduced slightly following perindopril (NS). Arterial wave reflections were significantly (P < 0.001) decreased with perindopril in comparison with atenolol, but this effect on wave reflections was not associated with a larger decrease in carotid artery PP. 5. Thus, during chronic treatment, ACE inhibition and selective beta1-adrenoceptor blockade resulted in a similar decrease in brachial and carotid artery PP, but only atenolol reduced heart rate. Aortic pulse wave velocity was reduced with both drugs, but atenolol appeared more effective in improving aortic stiffness. Arterial wave reflections were decreased only following perindopril. 6. Central pulse pressure was improved following 1 month treatment with an ACE inhibitor or beta-adrenoceptor blockade following a decrease in arterial wave reflections with perindopril and a higher decrease in regional aortic stiffness with atenolol. PMID- 11903320 TI - Recent developments concerning diet and hypertension. AB - 1. Recent data from randomized controlled dietary trials have shown blood pressure-lowering effects of foodstuffs and dietary patterns to be of practical importance for both individual and population blood pressure control. 2. The salient studies include Dietary Approaches to Stop Hypertension (DASH) trials, on complex dietary patterns and of additive effects of salt restriction, Trial of Nonpharmacologic Interventions in the Elderly (TONE), on weight control and sodium restriction as substitutes for drug therapy, and two Australian trials showing additive effects of dietary fish and weight control and of dietary protein and fibre in treated hypertensives. 3. Regular coffee drinking raised blood pressure in older hypertensives, whereas potential antihypertensive effects of dietary anti-oxidants require further scrutiny. PMID- 11903321 TI - Is primary aldosteronism underdiagnosed in clinical practice? AB - 1. Primary aldosteronism is a syndrome consisting of hypertension, suppressed renin activity or concentration and high aldosterone levels in plasma or urine. The main steps in diagnosis are the determination of renin and aldosterone levels, the demonstration of renin-aldosterone dissociation and discrimination between idiopathic hyperplasia and Conn's adenoma, with only Conn's adenoma amenable to surgery. 2. Patients with resistant hypertension and/or hypokalaemia should be screened for primary aldosteronism with simple, redundant hormonal tests. The aldosterone to renin ratio is a logical initial screening test, a high ratio demonstrating renin-aldosterone dissociation. Criteria for a high ratio should be determined in each laboratory. 3. In patients with documented primary aldosteronism, computed tomography scan and adrenal vein sampling help to distinguish between idiopathic hyperplasia and Conn's adenoma. 4. Patients with low renin hypertension, idiopathic hyperplasia and Conn's adenoma have overlapping values for plasma concentrations of potassium, renin and aldosterone and the aldosterone to renin ratio. Because primary aldosteronism subtypes are quantitative diseases, the true prevalence of primary aldosteronism cannot be defined. 5. The use of sensitive screening tests (e.g. aldosterone to renin ratio) gives a higher prevalence of diagnosed cases of primary aldosteronism, but not of surgically correctable forms. Therefore, there is no clinical evidence that primary aldosteronism is underdiagnosed. 6. There is a need for tests to predict the postoperative blood pressure outcome of surgery in subjects with Conn's adenoma. PMID- 11903322 TI - Familial varieties of primary aldosteronism. AB - 1. Improved approaches to screening and diagnosis have revealed primary aldosteronism (PAL) to be much more common than previously thought, with most patients normokalaemic. The spectrum of this disorder has been further broadened by the study of familial varieties. 2. Familial hyperaldosteronism type I (FH-I) is a glucocorticoid-remediable form of PAL caused by the inheritance of an adrenocorticotrophic hormone (ACTH)- regulated, hybrid CYP11B1/CYP11B2 gene. Diagnosis has been greatly facilitated by the advent of genetic testing. The severity of hypertension varies widely in FH-I, even among members of the same family, and has demonstrated relationships with gender, degree of biochemical disturbance and hybrid gene crossover point position. Hormone "day curve" studies show that the hybrid gene dominates over wild-type CYP11B2 in terms of aldosterone regulation. This may be due, in part, to a defect in wild-type CYP11B2-induced aldosterone production. Control of hypertension in FH-I requires only partial suppression of ACTH and much smaller glucocorticoid doses than previously recommended. 3. Familial hyperaldosteronism type II (FH-II) is not glucocorticoid remediable and is not associated with the hybrid gene mutation. Familial hyperaldosteronism type II is clinically, biochemically and morphologically indistinguishable from apparently non-familial PAL. Linkage studies in one informative family did not show segregation of FH-II with the CYP11B2, AT1 or MEN1 genes, but a genome-wide search has revealed linkage with a locus in chromosome 7. As has already occurred in FH-I, elucidation of causative mutations is likely to facilitate earlier detection of PAL. PMID- 11903323 TI - Progress in reducing the burden of stroke. AB - 1. The burden of stroke worldwide is growing rapidly, driven by an ageing population and by the rapid rate of urbanization and industrialization in the developing world. There are approximately 5 million fatal and 15 million non fatal strokes each year and over 50 million survivors of stroke alive, worldwide, today. 2. The most important determinant of stroke risk is blood pressure, with a strong, continuous relationship between the level of the systolic and diastolic pressures and the risk of initial and recurrent stroke, in both Western and Asian populations. 3. Randomized clinical trials have clearly demonstrated that blood pressure lowering reduces the risk of initial stroke by 35-40% in hypertensive patients; but, until recently, there was no conclusive evidence that blood pressure lowering was effective in the secondary prevention of stroke. 4. The Perindopril Protection Against Recurrent Stroke Study (PROGRESS) has provided definitive evidence that blood pressure lowering in patients with previous stroke or transient ischaemic attack (TIA) reduces the incidence of secondary stroke by 28%, of major vascular events by 26% and of major coronary events by 26%. These reductions were all magnified by approximately 50% in a subgroup of patients in whom the angiotensin-converting enzyme inhibitor perindopril was routinely combined with the diuretic indapamide. 5. Successful global implementation of a treatment with perindopril and indapamide in patients with a history of stroke or TIA would markedly reduce the burden of stroke and could avert between 0.5 and one million strokes each year, worldwide. PMID- 11903324 TI - Genetic influences on cardiovascular responses to an acoustic startle stimulus in rats. AB - 1. The aim of the present study was to assess the cardiovascular differences among five inbred rat strains (n=16 per strain), including spontaneously hypertensive rats (SHR), Wistar Kyoto (WKY) rats, Wistar Furth (WF) rats, Fischer (F344) rats and Lewis (Lew) rats and the usual outbred Wistar (W) rat strain (n=25). 2. These strains were compared under resting conditions for blood pressure (BP) and heart rate (HR) levels and for their baroreceptor-HR reflex sensitivity. In addition, their responses to an acoustic startle stimulus were measured. 3. A consistent rise in BP was observed among the groups as a result of the noise stimulus. This rise in systolic BP (SBP) averaged (+/-SEM) 37 +/- 2 mmHg in the SHR and 34 +/- 4 mmHg in F344 rats, while the response was only 23 +/ 3 mmHg in WKY rats. Pulse pressure (PP) was increased following noise in all groups. The delay for the BP response for all groups combined was 1.6 +/- 0.1 s. 4. Most animals had minimal HR variations, except F344 rats, responding with a 42 +/- 13 b.p.m. decrease 3.0 s after the stimulus (i.e. 1.3 s after the maximal 34 +/- 4 mmHg SBP rise). 5. The highest SBP (160 +/- 3 mmHg) and diastolic BP (104 +/- 3 mmHg) were observed in inbred SHR. Other groups were normotensive. Resting PP was elevated for SHR (56 +/- 2 mmHg) compared with the other groups (40 +/- 2 mmHg). The highest HR was found in F344 and WF rats, with 389 +/- 11 and 372 +/- 7 b.p.m., respectively. The lowest HR was observed in SHR and Lewis rats, with 335 +/- 7 and 323 +/- 7 b.p.m., respectively. The least sensitive baroreflex function was observed in SHR (0.8 +/- 0.1 b.p.m./mmHg) compared with the other strains (1.4 +/- 0.2 b.p.m./mmHg). 6. The present study confirms the importance of genetic factors on the cardiovascular responses of rats to a noise startle stimulus. Two inbred normotensive rat strains, namely F344 and WKY rats, which exhibit a substantial difference in pressor response to noise, may be used to unravel the mechanisms of sympathetic activation. PMID- 11903325 TI - Endothelin blockade in angiotensin II hypertension: prevention and treatment studies in the rat. AB - 1. The participation of endothelin (ET) in the development and maintenance of hypertension induced by angiotensin (Ang) II was assessed using the non-specific ET receptor antagonist bosentan (30 mg/kg per day). 2. In the prevention study, bosentan was given 24 h prior to and during the 17 day period of AngII infusion (200 ng/kg per min, s.c., osmotic pump), whereas in the treatment study, bosentan was administered from day 10 to day 17. Tail-cuff pressure (TCP) was measured before and on days 10 and 17 of AngII infusion. At the end of studies, heart weight index was calculated as the ratio of heart to bodyweight (HWI) and the wall thickness of the carotid artery (perfusion/fixation at 120 mmHg) was measured. Tail-cuff pressure increased from 129 +/- 3 to 179 +/- 7 and 189 +/- 9 mmHg on days 10 and 17 of AngII infusion, respectively. 3. Final TCP was markedly lowered in rats pretreated with bosentan, whereas TCP remained comparable with untreated hypertensive rats when bosentan was given from day 10 of AngII infusion (177 +/- 9 mmHg). 4. The increase in cardiac mass associated with AngII hypertension was similarly attenuated in the two groups receiving bosentan. 5. The HWI was 3.49 +/- 0.12 mg/g in untreated hypertensive rats and 3.18 +/- 0.08 and 3.11 +/- 0.09 mg/g in rats pretreated with bosentan and those receiving the antagonist from day 10. 6. The increase in carotid wall thickness induced by AngII was prevented, but not reversed, by bosentan. 7. These results support the hypothesis that endogenous ET participates only in the initial phase of AngII hypertension. In addition, the beneficial effect of bosentan of cardiac mass but not on arterial wall thickness is in favour of a role of ET as a local mediator of the cardiac hypertrophic effect of AngII, independently of the level of blood pressure and duration of hypertension. PMID- 11903326 TI - Impact of physical and physiological factors on arterial function. AB - 1. Arterial function measurements are increasingly used as surrogate markers of cardiovascular disease and it is important to define which non-pathological factors may influence these measurements. 2. The present study examined the influence of gender, height, body mass index (BMI), waist : hip ratio, heart rate and arterial pressure on pulse wave velocity (PWV), systemic arterial compliance (SAC) and central pressure augmentation index (AI) in 285 normal subjects, 98 males and 187 females, aged 50-82 years. 3. There were significant gender differences in PWV (higher in men), SAC (higher in men) and central pressure AI (lower in men). 4. Both SAC and AI were correlated with height in men and women and height largely accounted for gender differences. 5. Systemic arterial compliance was positively, whereas AI was negatively, correlated with BMI. 6. Both PWV and AI were significantly correlated with heart rate and central pulse pressure. 7. These findings may have implications for cardiovascular disorders. Reduced central arterial compliance and increased central pressure augmentation are potential mediators for the increased cardiovascular risk of short stature. A slow heart rate may contribute to increased central arterial pressure with potentially adverse consequences in older subjects. PMID- 11903327 TI - Blood pressure lowering in diabetes: a brief review of the current evidence and description of a new trial. AB - 1. Diabetes is a major global public health problem. The prevalence of this disease is predicted to increase sharply in the coming decades, particularly in less-developed regions of the world. 2. Most premature morbidity and mortality associated with diabetes relates to markedly increased risks of major cardiovascular diseases, such as myocardial infarction and stroke (macrovascular events), as well as microvascular complications, such as nephropathy and retinopathy. 3. Hypertension is a prevalent and important risk factor for vascular events in these patients. However, observational data demonstrate a continuous relationship between blood pressure and risk of vascular events, suggesting that even those individuals considered normotensive may benefit from blood pressure lowering. 4. Trials of blood pressure lowering among mostly hypertensive individuals with diabetes have demonstrated benefit of intervention on macrovascular and microvascular outcomes. Recent data may suggest additional effects of angiotensin- converting enzyme inhibitors independent of blood pressure lowering. 5. Issues where data are lacking with respect to blood pressure lowering in diabetes include the effects of blood pressure lowering among non-hypertensive individuals and the effects of blood pressure lowering regimens based on different classes of drug. 6. Data expected to address some of these issues are being collected. These include a prospective meta-analysis of blood pressure-lowering trials with large numbers of patients with diabetes. A new large-scale randomised trial, ADVANCE (Action in Diabetes and Vascular Disease), is also described. PMID- 11903328 TI - Inhibitory effects of angiotensin II on barosensitive rostral ventrolateral medulla neurons of the rat. AB - 1. The brain renin-angiotensin system can influence arterial baroreceptor reflex control of blood pressure (BP) through both direct and indirect effects on sympathetic premotor neurons of the rostral ventrolateral medulla (RVLM). The present study examined the direct effect of angiotensin (Ang) II applied by microiontophoresis on the ongoing activity of single RVLM neurons. 2. In 26 urethane-anaesthetized Wistar rats, recordings of single unit activities of barosensitive RVLM neurons were made from one barrel of a six-barrel micropipette assembly. The other five barrels were filled with either L-glutamate, AngII, valsartan (an AT1 receptor antagonist), PD 123177 (an AT2 receptor antagonist) and saline. All drugs were applied by microiontophoresis. 3. Mean BP was 83 +/- 3 mmHg. Application of AngII inhibited the ongoing activity of RVLM neurons, identified as barosensitive because their activity was inhibited by a phenylephrine- induced increase in BP, from 12.6 +/- 1.5 to 5.4 +/- 1.1 Hz (n=24; P < 0.001). Angiotensin II also inhibited the glutamate-evoked excitation of barosensitive RVLM neurons from 15 +/- 3 to 5.8 +/- 2.0 Hz (n=6; P < 0.001). Valsartan significantly increased neuronal activity from 9.5 +/- 2.3 to 13.5 +/- 3.2 Hz (n=7, P < 0.01), whereas PD 123177 significantly decreased neuronal activity from 13.5 +/- 3.5 to 9.9 +/- 2.8 Hz (n=13; P < 0.01). 4. The results suggest that AngII exerts a tonic inhibitory effect on barosensitive RVLM neurons, which is presumably mediated through AT1 receptor stimulation. PMID- 11903330 TI - Right place-right time; real time protein-protein interactions. PMID- 11903331 TI - Shutting off mammalian gene expression the easy way. PMID- 11903329 TI - Goals of genetic counseling. AB - The goals of genetic counseling have differed over the past three decades. Two schools of thought are prominent in reviewing past literature. One upholds the goal of preventing birth defects and genetic disorders while the other promotes a goal of improved psychological well-being in client adaptation to a genetic condition or risk. Both types of goals emphasize that clients should make their own reproductive decisions; however, the former relies on clients making decisions that will reduce the impact of genetic disorders. The differences in the types of goals may be due to the training and orientation of genetics health care providers, socio-cultural views, or priorities of health care settings. Regardless, there are ample reasons to dismiss the prevention of birth defects as a goal. This mini-review recommends use of genetic counseling sub-specialties as a framework for considering different client needs and thus different counseling goals and specific aims in the reproductive, pediatric/adult, and common disease settings. Given the extent of new genetic information, technologies, and the need to evaluate genetic counseling practice, genetics health care providers should work toward arriving at consensus on the goals of genetic counseling, and in doing so, the needs of clients should be considered. PMID- 11903332 TI - Genetic studies yield conflicting data in schizophrenia. PMID- 11903333 TI - Trisomy 17p10-p12 resulting from a supernumerary marker chromosome derived from chromosome 17: molecular analysis and delineation of the phenotype. AB - We report a 5-year-old boy with a small de novo marker chromosome derived from the proximal short arm of chromosome 17. His clinical features include hypotonia, global developmental delay, oval face with large nose and prominent ears, and ligamentous laxity of the fingers. Magnetic resonance imaging of the brain demonstrated mildly delayed myelination. G-band chromosome analysis revealed mosaicism for a small marker chromosome in 85% of the peripheral blood cells analyzed. Fluorescence in situ hybridization and microsatellite polymorphism studies showed that the der(17) was of maternal origin and included genetic material from the 17p10-p12 region, but did not contain the PMP22 gene. One breakpoint mapped within the centromere and the second breakpoint mapped adjacent to the Charcot-Marie-Tooth disease type 1A proximal low-copy repeat (CMT1A-REP). We compare the clinical characteristics of our patient with those previously reported to have a duplication involving the proximal short arm region of chromosome 17 to further delineate the phenotype of trisomy 17pl0-p12. PMID- 11903334 TI - Association of a single nucleotide polymorphism in CPB2 encoding the thrombin activable fibrinolysis inhibitor (TAF1) with blood pressure. AB - Thrombin-activable fibrinolysis inhibitor (TAFI) is a hepatically secreted zymogen, whose substrates include bradykinin. The CPB2 gene encoding TAFI is a candidate gene for blood pressure. A recently identified single nucleotide polymorphism (SNP) in the CPB2 coding region, designated as 1057C > T, results in an amino acid change at TAFI residue 325 (Ile > Thr325). We found that the genotype based on this SNP was significantly associated with blood pressure in aboriginal Canadians. Specifically, analysis of variance showed that homozygotes for CPB2 1057T had significantly lower diastolic blood pressure than subjects with other CPB2 genotypes. CPB2 genotype accounted for approximately 3% of the total variation in diastolic blood pressure. consistent with the expected magnitude of a modest genetic effect in a complex trait such as blood pressure. Although the mechanism underlying the association is unclear, the findings are of interest because TAFI may provide a link between coagulation and blood pressure regulation. PMID- 11903335 TI - A PCR-based method for detecting known mutations in the human UDP galactose-4' epimerase gene associated with epimerase-deficiency galactosemia. AB - Epimerase-deficiency galactosemia results from impairment of the human enzyme UDP galactose-4'-epimerase (GALE). We report a rapid, internally controlled PCR-based method for detection of nine naturally occurring point mutations in human GALE associated with epimerase deficiency. These mutations were derived from patients whose clinical presentations ranged from mild to severe; all but one of these mutations have been reported previously. The tests described here work well on both cDNA and genomic samples and require no specialized equipment beyond a thermal cycler and an agarose gel electrophoresis system. Finally, although these tests in no way replace the need for biochemical diagnosis in epimerase deficiency galactosemia, they do provide the possibility of additional molecular information to support a biochemical diagnosis and facilitate the possibility of more accurate carrier testing, should that option be desired. PMID- 11903336 TI - Familial case of Potocki-Shaffer syndrome associated with microdeletion of EXT2 and ALX4. AB - Multiple exostosis, biparietal foramina, minor craniofacial abnormalities, and mental retardation are characteristic of the syndrome associated with a proximal deletion of 11p (MIM # 601224), which has been shown to be a true contiguous gene deletion syndrome. The presence of multiple exostosis is associated with deletion of the EXT2 gene. Similarly, the presence of biparietal foramina has been shown to be associated with the deletion of ALX4 located proximally to EXT2. Specific genes related to mental retardation and craniofacial abnormalities, however, have yet to be identified. We report on a family with a microdeletion of 11(pll.2p11.2) with multiple exostosis and biparietal foramina without mental retardation or craniofacial abnormalities. Our results suggest that genes related to mental retardation and craniofacial development must be located outside of the D11S1785-D11S1385 region. PMID- 11903337 TI - Evaluation for sleep apnea in patients with Ehlers-Danlos syndrome and Marfan: a questionnaire study. AB - Sleep complaints are frequently reported by patients with Marfan and Ehlers Danlos syndrome (EDS). We examined the exact nature of sleep complaints in these patients. A representative sample of Marfan and EDS patients responded to a general sleep questionnaire, including the Epworth Sleepiness Scale (ESS) and the Medical Outcomes Study Short-Form 36 (SF-36) health-related quality of life (QOL) questionnaire. Fifteen Marfan patients and 9 EDS patients were evaluated and compared to 24 healthy controls, matched for age, sex and body mass index. Maintaining sleep was frequently disturbed in Marfan (40%, p < 0.04) as well as in EDS patients (56%, p < 0.01). Sleep apnea was exclusively reported by Marfan patients (27%, p = 0.03). Periodic limb movements were much more reported in EDS (67%, p = 0.02) than in Marfan (27%, p = 0.25) compared to controls (8%). Pain and back complaints were highly presented in both groups, but most pronounced in EDS patients (47% in Marfan versus 77% in EDS). No differences for the scores in the ESS were found. For all SF-36 questionnaire items, scores were much lower in patient groups, except for emotional problems. We found that sleep complaints were not rare in Marfan and EDS patients and correlated well with different QOL items. Our study calls for greater attention to the presence of apnea, pain and periodic limb movements in these patients. PMID- 11903338 TI - Tandem duplication mosaicism: characterization of a mosaic dup(5q) and review. AB - Mosaicism for tandem duplications is rare. Most patients reported had abnormal phenotypes of varying severity, depending on the chromosomal imbalance involved and the level of mosaicism. Post-zygotic unequal sister-chromatid exchange has been proposed as the main mechanism for tandem duplication mosaicism. However, previous molecular analyses have implicated both meiotic and post-zygotic origins for the duplication. We describe a newborn male who was originally diagnosed in utero with arrhythmia and tetralogy of Fallot. He had multiple dysmorphic features including telecanthus, blepharophimosis, high broad nasal bridge with a square-shaped nose, flat philtrum, thin upper lip, down-turned corners of the mouth, high-arched palate, micrognathia, asymmetric ears, and long, thin fingers and toes. Karyotyping of peripheral blood lymphocytes showed mosaicism for a tandem duplication of part of the long arm of one chromosome 5: mos46,XY,dup(5)(q13q33)[6]/46,XY[45]. Fibroblast cultures had the same mosaic karyotype with a higher frequency of the dup(5) clone: mos46,XY,dup(5)(q13q33)[9]/46,XY[21]. Fluorescence in situ hybridization analysis with a wcp5 confirmed the chromosome 5 origin of the additional material. Parental karyotypes were normal indicating a de novo origin of the dup(5) in the proband. Molecular analyses of chromosome 5 sequence-tagged-site (STS) markers in our family were consistent with a post-zygotic origin for the duplication. Therefore, mosaicism for tandem duplications can arise both through meiotic or mitotic errors, as a result of unequal crossing over or unequal sister-chromatid exchange, respectively. Our review indicates that mosaicism for tandem duplications is likely under-ascertained and that parental karyotyping of probands with non-mosaic tandem duplications should be performed. PMID- 11903339 TI - Familial deletion of (8)(q24.13q24.22) associated with a normal phenotype. AB - We report a familial deletion of (8q) detected in amniocytes of a fetus with a normal ultrasound and in the phenotypically normal mother, who has now had three pregnancy losses. Chromosome analysis of amniocytes and maternal peripheral blood cells showed an interstitial deletion of (8)(q24.13q24.22), which is distal to the region associated with Langer-Giedion syndrome (LGS) or trichorhinophalangeal (TRP) syndrome. This deletion was confirmed by fluorescence in situ hybridization with a c-myc cosmid clone and chromosome 8 painting library. PMID- 11903340 TI - Lack of association of the common TaqIB polymorphism in the cholesteryl ester transfer protein gene with angiographically assessed coronary atherosclerosis. AB - The anti-atherogenic effect of cholesteryl ester transfer protein (CETP) genetic variants associated with lowered enzyme activity is controversial. Moreover, in a few studies, this effect has been evaluated in the presence of a certain risk factor constellation. We addressed this issue in a case-control study, where 415 subjects with angiographically documented coronary artery disease (CAD +), 397 subjects without CAD (in 215, CAD was excluded by coronarography (CAD-)), and 188 healthy population controls, were screened for the CETP TaqIB polymorphism. The prevalence of the low-activity TaqIB2 allele was 0.396 in CAD+, and 0.428 and 0.416 in CAD- and population controls, respectively (p = 0.40). Its presence was significantly associated with increased high-density lipoprotein cholesterol (HDL C) in population controls (1.40 +/- 0.40 mmol/l in B1B1, 1.52 +/- 0.39 mmol/l in B1B2 and 1.58 + 0.46 mmol/l in B2B2; p < 0.03 for trend), but not in the other groups. The CETP TaqIB polymorphism accounted for < 1% of the HDL-C variance in the whole cohort (p = 0.048). After adjustment for other risk factors, the CETP TaqIB2 allele was found not to be associated with significant changes in CAD risk independently of an assumed either dominant (odds ratio (OR) 0.97; 95% confidence interval (CI) 0.66-1.44; p = 0.89) or recessive effect (OR 0.68; 95% CI 0.42 1.12; p = 0.13). The CETP TaqIB polymorphism did not show a significant interaction with other risk factors in influencing CAD risk. Our findings do not support the hypothesis that a genetic variant resulting in lowered CETP activity is associated with reduced risk of coronary atherosclerosis. PMID- 11903341 TI - Apolipoproteins AI, B, and E polymorphisms in severe aortic valve stenosis. AB - Hypercholesterolemia has been related to aortic valve stenosis (AS). Polymorphisms of apolipoproteins (apo) AI, B, and E are associated with variable levels of plasma lipids, but the association between these polymorphisms and AS is unknown. In a case-control study of groups matched by age, sex, comparable body mass index, hypertension, triglycerides, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol, we analyzed the distribution of apo AI A/G mutation, apo B signal peptide insertion/deletion, apo B XbaI restriction fragment length. and apo E polymorphisms in 62 non-diabetic patients with severe aortic valve stenosis and 62 control subjects. All patients underwent echocardiographic analysis. Univariate analysis showed a higher prevalence of the XbaI X + /X + genotype (p = 0.007) of apo B and the apo E2 allele (p = 0.034) in patients with severe AS. Apo polymorphisms were not associated with lipid levels, left ventricular mass, or the aortic gradient. PMID- 11903342 TI - PCR-PRINS-FISH analysis of structurally abnormal sex chromosomes in eight patients with Turner phenotype. AB - According to cytogenetic analysis, about 50% of Turner individuals are 45,X. The remaining cases have a structurally abnormal X chromosome or are mosaics with a second cell line containing a normal or abnormal sex chromosome. In these mosaics, approximately 20% have a sex marker chromosome whose identity cannot usually be determined by classical cytogenetic methods, requiring the use of molecular techniques. Polymerase chain reaction (PCR), primed in situ labeling (PRINS), and fluorescence in situ hybridization (FISH) analyses were performed in 8 patients with Turner syndrome and 45,X mosaic karyotypes to determine the origin and structure of the marker chromosome in the second cell line. Our data showed that markers were Y-derived in 2 patients and X-derived in the remaining 6 patients. We were also able to determine the breakpoints in the two Y chromosomes. The use of cytogenetic and molecular techniques allowed us to establish unequivocally the origin, X or Y, of the marker chromosomes in the 8 patients with Turner phenotype. This study illustrates the power of resolution and utility of combined cytogenetic and molecular approaches in some clinical cases. PMID- 11903344 TI - Segmental uniparental disomy of 7q31-qter is rare in Silver-Russell syndrome. PMID- 11903345 TI - The VACTERL association: lessons from the Sonic hedgehog pathway. PMID- 11903346 TI - A Reply. PMID- 11903343 TI - The frequency of common mutations among patients with mucopolysaccharidosis types I, II and IIIA diagnosed in Austria over the last 17 years. PMID- 11903347 TI - Genetic services. AB - This is a time of unprecedented increase in knowledge about the genetic basis of disease against a background of rapidly changing technology. Advances happen quickly, with the new knowledge rapidly becoming relevant to services for patients, and hence there is an increasing demand for, and expectation of, genetic services. This brings a challenge for health services worldwide to keep pace with the expectations of their populations. There is also a need for public and professional education and dialogue to dispel some of the hype and myths about what can be achieved. It should not be forgotten that any services must be provided and developed within a broad ethical framework. A statement from a WHO expert consultation (1) concluded that 'Genetic advances will only be acceptable if their application is carried out ethically, with due regard to autonomy, justice, education and the beliefs and resources of each nation and community'. That so many public bodies have commented on genetic services and their associated ethical, legal and social issues emphasizes not only the importance of this area in health care, but also that the wider issues are of great importance in democratic societies. The aim of this paper is to review the current situation in the provision of genetic services, to examine the drivers for change, to speculate on the likely need for future services and to suggest models for the development of such services. Pharmacogenetics and the determination of genetic factors indicating susceptibility to infection will not be discussed since these have been the subject of several recent excellent reviews (2, 3). Genetic services have been defined as health measures implemented to help people with a genetic disadvantage and their families to live and reproduce as normally as possible (4). Broadly they can be divided into those services which target whole populations with a view to identifying those at increased risk; and those which focus on the needs of families which are affected by a genetic disorder or who perceive themselves to be at increased risk. PMID- 11903348 TI - HotSpots. PMID- 11903349 TI - Standardization of PCR amplification for fragile X trinucleotide repeat measurements. AB - To provide the clinical diagnostics community with accurate protocols and measurements for the detection of genetic disorders, we have established a quantitative measurement program for trinucleotide repeats associated with human disease. In this study, we have focused on the triplet repeat associated with fragile X syndrome. Five cell lines obtained from the Coriell Cell Repository were analyzed after polymerase chain reaction (PCR) amplification and size separation. These cell lines were reported to contain CGG repeat elements (ranging from 29 to 110 repeats). Our initial measurements focused on measurement variability: (a) between slab-PAGE and capillary (CE) separation systems (b) interlane variability (slab-PAGE) (c) intergel variability, and (d) variability associated with amplification. Samples were run in triplicate for all measurements, and the analysis performed using Gene Scan analysis software. The repeat sizes were verified by DNA sequence analyzes. The standard deviations for interlane measurements in slab-gels ranged from 0.05 to 0.35. There was also little variation in size measurements performed on different gels and among PCR amplifications. The CGG repeat measurements performed by capillary electrophoresis were more precise, with standard deviations ranging from 0.02 to 0.29. The slab-PAGE and CE size measurements were in agreement except for the pre mutation alleles, which yielded significantly smaller sizes by CE. PMID- 11903351 TI - Telomere-specific fluorescence in situ hybridization analysis of couples with five or more recurrent miscarriages. AB - Fluorescence in situ hybridization analysis using telomere specific probes has been used to detect cryptic translocations in the chromosomal telomeric regions. This study was performed in five clinically normal couples who have had five or more spontaneous abortions and whose karyotypes were found to be normal using conventional cytogenetic techniques. Using the telomere specific probes, in one couple we determined a cryptic translocation between chromosome 3 and 10, and, in another couple, the signal in chromosome 20 was detected in another chromosome, which was probably a D group chromosome. Additionally, in the latter and also in two other couples, we observed a polymorphism. The approach will be helpful for screening cryptic translocations using telomere specific multiple probe sets in couples with recurrent miscarriages. As prenatal diagnosis will be available for these couples for future pregnancies, it will be possible to help these families to have healthy fetuses. PMID- 11903350 TI - A prospective evaluation of the angiotensin-converting enzyme D/I polymorphism and left ventricular remodeling in the 'Healing and Early Afterload Reducing Therapy' study. AB - The D/I (deletion, D, insertion, I) polymorphism of the angiotensin-converting enzyme (ACE) gene has been extensively studied for its association with a number of cardiovascular and other disease states. However, its potential association with differential clinical efficacy of ACE inhibitors (ACE-I) administered to patients who had suffered a myocardial infarction (MI), i.e. the prevention of left ventricular (LV) remodeling, has so far not been specifically studied. The aim of the study was to investigate whether the D/I polymorphism of the ACE gene is associated with the incidence of post-MI LV remodeling in patients drawn from the 'Healing and Early Afterload Reducing Therapy' (HEART) Study. The ACE D/I polymorphism was characterized by the polymerase chain reaction (PCR) in 265 subjects from the 'Healing and Early Afterload Reducing Therapy' Study, a double blind, placebo-controlled trial with the objective of determining whether early or delayed administration of the ACE-I, ramipril, in patients with acute anterior wall MI would be optimal in reducing LV enlargement. Selected frequencies for the ACE D and I alleles were 0.59 and 0.41 (placebo-high dose group), 0.56 and 0.44 (low dose-low dose group), and, 0.60 and 0.40 (high dose-high dose group), respectively. All observed genotype frequencies were in Hardy-Weinberg equilibrium. There was no evidence for an association between genotype and outcome regarding LV size or function, nor with the initial blood pressure response after ACE-I administration (adjusted for covariates). Our data provide no evidence for an association of the ACE D/I polymorphism with the risk of LV remodeling post-MI in the presence of ACE-I therapy, and therefore do not suggest that differential clinical efficacy of ACE-inhibitors is related to this genetic marker. PMID- 11903352 TI - The E326K mutation and Gaucher disease: mutation or polymorphism? AB - Gaucher disease is caused by mutations in the gene for human glucocerebrosidase, a lysosomal enzyme involved in the intracellular hydrolysis of glucosylceramide. While over 150 different glucocerebrosidase mutations have been identified in patients with Gaucher disease, not all reported mutations have been fully characterized as being causative. One such mutation is the E326K mutation, which results from a G to A nucleotide substitution at genomic position 6195 and has been identified in patients with type 1, type 2 and type 3 Gaucher disease. However, in each instance, the E326K mutation was found on the same allele with another glucocerebrosidase mutation. Utilizing polymerase chain reaction (PCR) screening and restriction digestions of both patients with Gaucher disease and normal controls, we identified the E326K allele in both groups. Of the 310 alleles screened from patients with Gaucher disease, the E326K mutation was detected in four alleles (1.3%). In addition, screening for the E326K mutation among normal controls from a random population revealed that three alleles among 316 screened (0.9%) also carried the E326K mutation. In the normal controls with the E326K allele, the glucocerebrosidase gene was completely sequenced, but no additional mutations were found. Because the E326K mutation may be a polymorphism, we caution that a careful examination of any allele with this mutation should be performed to check for the presence of other glucocerebrosidase mutations. PMID- 11903353 TI - Fertility and pregnancy outcome in Danish women with Turner syndrome. AB - The aim of the investigation was to study fertility in Danish women diagnosed with Turner syndrome (TS), and to describe their offspring. In total, 410 women in the fertile age were registered in the Danish Cytogenetic Central Register with TS between January 1973 and December 1993. Their karyotype were as follows: 49% with 45,X, 23% with mosaicism and a structural abnormality of the second X, 19% with 45,X/46,XX mosaicism, and 9% with 46,XX and a structural abnormality of the second X. Thirty-three women, one with 45,X, 27 with mosaicism and 5 with 46,XX and a structural abnormality of the second X, gave birth to 64 children. Two women had become pregnant after in vitro fertilization, including a woman with 45,X after an egg donation. Thus, 31 women(7.6%) had achieved at least one spontaneous pregnancy, but 48% of the fertile women registered with 45,X/46,XX had 45,X in less than 10% of the analysed cells. Twenty-five of the 64 children had a chromosome analyzed. Six of the 25 examined children, including three siblings, had chromosomal aberrations. No case of Down's syndrome was present, and only two children had malformations. Fertility in women registered with TS is higher than earlier reported. However, only women with 45,X/46,XX mosaicism or 46,XX and structural abnormality of the second X, gave birth to live children after spontaneous pregnancies. PMID- 11903354 TI - A previously undescribed nonsense mutation of the HFE gene. AB - A patient with clinically manifest hereditary hemochromatosis was found to be heterozygous for the c.845 A-->G (C282Y) mutation. As simple heterozygotes for this mutation do not develop the hemochromatosis phenotype, the coding region of the patient's HFE gene was sequenced and a previously undescribed nonsense mutation was identified at c.211 C-->T (R74X). The patient's brother who also had the hemochromatosis phenotype shared his HFE genotype. To determine how common such mutations might be, the coding and 5' region of the HFE genes of 11 subjects who had been found in a large population survey to be heterozygous for the C282Y mutation and had elevated ferritin levels were sequenced. No mutations were found. Sequencing of the HFE gene also revealed two polymorphisms that had not previously been noted, -467 C-->G and -970 T-->G. Neither of these mutations appear to cause an abnormality in iron metabolism. PMID- 11903355 TI - Distribution of HFE C282Y and H63D mutations in the Balearic Islands (NE Spain). AB - The HFE gene contains two main missense mutations: C282Y and H63D. Individuals with these mutations carry a risk of developing hereditary haemochromatosis (HH). The common form of this disease is due to homozygosity for the C282Y mutation. Population studies have shown the variation of the prevalence of these mutations in different countries and ethnic groups. The purposes of this current study were to determine the prevalence of the C282Y and H63D mutations in the Balearic Islands and the genotypic characterization of patients diagnosed with HH, as well as those with iron overload and liver diseases. A total of 1330 Balearic chromosomes were analyzed. The results showed that the populations of the Balearic Islands were not homogeneous. No C282Y carriers were observed in a group of descendants of Majorcan Jews (Chuetas) and the frequency was very low in Minorca (1.2%) in comparison with the other islands of Majorca (4.7%) and Ibiza (6.5%). The carrier frequency of the H63D mutation was similar in the three islands and very high (43.1%) in the descendants of Majorcan Jews. The study of patients was carried out in 129 individuals. The homozygous C282Y genotype was the principal one involved in hereditary haemochromatosis (90%), whereas the other HH patients were C282Y/H63D compound heterozygous and H63D homozygous. PMID- 11903356 TI - Klinefelter syndrome is a common cause for mental retardation of unknown etiology among prepubertal males. AB - Klinefelter syndrome (KS) has not typically been associated with mental retardation (MR), however, in recent years a growing body of evidence suggested that KS boys often experience language deficits and academic difficulties. In this study, we screened DNA samples from 1205 patients originally referred for fragile X syndrome (FRAX) testing, because of MR of unknown etiology and detected 8 KS patients. A similar number of males in the same age group were found to have FRAX; 3 of them had a family history of FRAX. Based on these findings, KS might be the most common cause of MR of unknown etiology among prepubertal males. Because of the significant benefits of early recognition and treatment of KS, we emphasize the importance of cytogenetic testing of all prepubertal males with cognitive impairment even without dysmorphic features. PMID- 11903357 TI - Partial Xp duplication in a girl with dysmorphic features: the change in replication pattern of late-replicating dupX chromosome. AB - In this paper we present the case of a girl at the age of 32 months with dysmorphic features, including general muscular hypotonia, developmental delay and mental retardation. The cytogenetic analysis revealed de novo partial duplication of Xp: 46,X,dup(X)(p11.23-->p22.33: :p11.23-->p22.33). To characterize the duplication, X painting, Kallman (KAL), yeast artificial chromosomes (YACs) and bacterial artificial chromosomes (BACs) covering Xp11.23- >Xp22.33 region were used. Selective inactivation of the abnormal X chromosome using HpaII digestion of the AR gene was evident. After BrdU incorporation the abnormal X was late-replicating in all lymphocytes examined. There was one peculiar exception observed: the break-point region was consistently early replicating. The replicating pattern of this region corresponded to the active X chromosome. Methylation pattern of late replicating X chromosome was studied also using antibodies against 5-methylcytosine. The pattern corresponded to the normally inactive X chromosome, with the exception of the previously observed break-point region which revealed an early replicating pattern with strong fluorescent signal, similar to the pattern of the active X chromosome. The observed phenomenon could lead to the abnormal phenotype of the patient, with some normally inactive genes of the break-point region escaping the inactivation process. The abnormal clinical findings could also be due to tissue-dependent differences in the inactivation pattern. PMID- 11903358 TI - Tetraploidy in a 26-month-old girl (cytogenetic and molecular studies). AB - Liveborn infants with tetraploidy are very rare in human pregnancies and usually die during the first days or months. Seven cases of liveborn infants with tetraploidy have previously been reported. Among them only two 92, XXXX infants survived for longer than 12 months. Here we report on the case of a 26-month-old girl with tetraploidy. The main clinical features of tetraploidy are facial dysmorphism, severely delayed growth and developmental delay. On the basis of molecular studies we discuss the possible origin of the additional chromosome sets in our proband. To our knowledge, this infant is the first reported case of tetraploidy who lived up to 26 months. PMID- 11903359 TI - Association of a polymorphism of the ecNOS gene with myocardial infarction in a subgroup of Turkish MI patients. AB - In this study we examined a possible association between a 27 base pair (bp) repeat polymorphism in intron 4 of the ecNOS gene and myocardial infarction (MI) in a subgroup of the Turkish population. We compared MI and control groups for the frequencies of the ecNOS alleles and their genotypes. The frequency of the ecNOS 4a/a and 4a/b genotypes was found to be significantly higher in the MI group than in the control group. Interestingly, the frequency of the ecNOS 4a/b polymorphism was found to be significantly higher in the selected MI group (patients with no known secondary risk factors) than in the control and non selected MI group. We found that the patients with MI had the frequency of the a/a genotype 4.3%, of the a/b genotype 26.6% and the b/b genotype 69.1%. The controls, however, showed only 0.6% for a/a, 18.0% for a/b and 81.4% for the b/b genotype (P < 0.001; chi2 = 13.626). In this study, we show that myocardial infarction is associated with one subtype of ecNOS gene polymorphism. PMID- 11903360 TI - Familial Mediterranean fever: the potential for misdiagnosis of E148V using the E148Q usual RFLP detection method. PMID- 11903361 TI - Autosomal dominant isolated velopharyngeal insufficiency. PMID- 11903362 TI - A Japanese patient with cerebrotendinous xanthomatosis has different mutations within two functional domains of CYP27. PMID- 11903363 TI - The role of antibiotics in the management of acute otitis media in children. PMID- 11903364 TI - The Audioscan: a high frequency resolution audiometric technique and its clinical applications. AB - The Audioscan is a form of high definition audiometry based on iso-hearing level frequency sweeps, which was developed by Meyer-Bisch in 1990. Compared with traditional tone audiometry, it sweeps across the preset frequency range at a predetermined sweep rate and provides a continuous audiometric curve. Because the device has a maximum frequency range of 125-16 000 Hz with 64 frequencies per octave, the Audioscan method can, theoretically, give 64 times as many values as fixed-frequency audiometry, which may give greater accuracy and sensitivity. An advantage of this is its capacity to detect mild audiometric deficits such as narrow notches situated between the frequencies normally tested. These may represent very limited auditory lesions, at a stage when they cannot be detected by routine audiological methods. Thus, the Audioscan method can give not only a detailed audiometric curve, but also provide important indicators of mild auditory dysfunction. The Audioscan device (e.g. Essilor model) is commercially available for clinical purpose. It is a software-based system, which can also be used for pure tone audiometry and Bekesy audiometry. This paper reviews the general aspects of the Audioscan technique and current applications for detecting auditory dysfunction. This would be valuable to provide some guidelines on the Audioscan assessment, and contribute to a clarification of the clinical application of Audioscan and facilitate further research. PMID- 11903365 TI - CT of the paranasal sinuses: a review of the correlation with clinical, surgical and histopathological findings. AB - Computerized tomography (CT) of the paranasal sinuses provides valuable information but this should be interpreted in the context of the history and examination as the prevalence of incidental mucosal changes in an asymptomatic population is approximately 30%. A review of the presence or extent of the various anatomical variations that are found in the paranasal sinuses does not differ between a symptomatic and an asymptomatic population. This makes it unlikely that these are very important in either initiating or sustaining paranasal sinus disease. CT provides an excellent map to help the sinus surgeon operate. CT provides information about the extent of mucosal disease but this correlates poorly with symptoms, surgical findings and histopathology. CT does provide invaluable information to help in the diagnosis of atypical sinus infections, malignancy and in the management of the complications of rhinosinusitis. A normal CT in a patient with facial pain should make the doctor consider another diagnosis. In essence, CT helps to support a clinical diagnosis but it should not be interpreted out of context, and it is therefore vital that doctors communicate the clinical picture to their radiological colleagues, and that they learn to interpret the radiographs. PMID- 11903367 TI - Clinical and pathological characteristics of oral lichen planus in hepatitis C positive and -negative patients. AB - The reported prevalence rate of anti-hepatitis C virus (HCV) antibodies in patients with oral lichen planus shows wide geographical variation and ranges from 0 to 65%. Certain characteristic clinical features have been attributed to oral lichen planus associated to HCV infection. The purpose of this investigation has been to assess hypothetical clinical differences, as well as differences in the intensity of the subepithelial inflammatory infiltrate between oral lichen planus-HCV +ve patients and oral lichen planus-HCV -ve patients. A total of sixty two patients entered the study. Their mean age was 63.5 +/- 14.49 years, and 48.4% of them were men and 51.6% women. Patients were classified according to their serum HCV positivity. Age, sex, clinical presentation (reticular or atrophic-erosive), extension of the lesions, location of the lesions, number of locations affected, intensity of the inflammatory infiltrate and Candida albicans colonization were recorded for each patient. Reticular lichen planus was the most frequent clinical presentation in both HCV +ve (57.1%) and HCV -ve patients (63.6%). C. albicans colonization ranged from 42.8% in HCV +ve and 41.7% in HCV ve patients. HCV + ve patients showed certain oral locations more frequently affected than HCV -ve ones: lip mucosa, 28.6% versus 7.3%; tongue, 57.1% versus 29.1%; and gingiva, 71.4% versus 23.6%. The number of affected intraoral locations was higher in HCV +ve patients (71.4%) than among HCV -ve ones (20.4%; chi2 = 8.34; P < 0.011). No statistically significant differences could be established in terms of density of subepithelial inflammatory infiltrate between the groups. Our results reinforce the need for liver examination in all patients with oral lichen planus, particularly those showing lesions on the gingiva with multiple intraoral locations affected, as no pathological differences could be identified between HCV + ve and HCV -ve patients. PMID- 11903366 TI - An evaluation of the best head position for instillation of steroid nose drops. AB - Steroid nose drops are used frequently to treat rhinosinusitis and nasal polyposis. The middle meatal area is of key importance in the pathophysiology of these conditions. This study assesses which of three head positions commonly used to instil nose drops resulted in the highest coverage of this area. Discomfort levels were also studied using a visual analogue scale for each position. Five volunteers were studied in: (i) head back (HB); (ii) head forward and down (HFD); and (iii) lying head back (LHB) positions. Betamethasone nose drops, dyed with fluorescein, were instilled into each nostril and the distribution was studied endoscopically. The middle meatus area distribution was highest in the LHB position (55.51%), followed by HFD (31.55%) and HB (6.87%) positions. Comparison of distribution between HB and LHB (P = 0.002) and between HB and HFD (P = 0.045) was statistically significant. The HFD position was most uncomfortable (mean discomfort score 8.8) whereas the HB (2.4) and LHB (2.6) were similar. The LHB would, therefore, appear to be the most suitable position for instillation of steroid nose drops. PMID- 11903368 TI - Is electronystagmography of diagnostic value in the elderly? AB - We aimed to determine the diagnostic value of electronystagmography (ENG) in a community-based sample of dizzy subjects over 65 years old. A total of 96 asymptomatic controls and 149 dizzy subjects underwent ENG. Clinical diagnoses were made on standardized criteria. ENG results were classed as normal or abnormal, according to reference ranges derived from the controls. Rates of ENG abnormality in different diagnostic categories, sensitivities, specificities and predictive values were calculated. Central vascular disease was common (105 out of 149 subjects); peripheral vestibular disease was not (14). Spontaneous nystagmus had a positive predictive value of 95% for central vascular disease, but was only 18%-sensitive, and was usually detectable clinically. ENG had no other significant diagnostic value. ENG failed to discriminate dizzy subjects from controls and failed to differentiate various dizziness syndromes. ENG was of no practical value in this community-derived sample of dizzy elderly subjects. PMID- 11903369 TI - Circulating angiogenic cytokines as tumour markers and prognostic factors in head and neck squamous cell carcinoma. AB - This pilot study investigated the potential use of three circulating angiogenesis related cytokines, basic fibroblast growth factor (bFGF), angiogenin (ANG) and endostatin, as tumour markers and prognostic factors in patients with head and neck squamous cell carcinoma (HNSCC). A total of 30 patients with HNSCC treated with curative intent and 15 healthy controls were studied. Serum (bFGF and ANG) and plasma (endostatin) was assayed by enzyme-linked immunoabsorbance assay (ELISA). None of the cytokines was raised in HNSCC patients when compared with controls. Serum bFGF was not associated with any clinico-pathological or outcome parameters, although there was a trend towards higher levels in more advanced and aggressive tumours. Lower serum angiogenin (sANG) levels were associated with loco-regional disease recurrence (P = 0.036). Using a cut-off level of 400 pg/mL, a low level of sANG predicted tumour recurrence with a relative risk of 4.0 (95% CI: 0.7-24.0). Plasma endostatin was associated with higher histological grade (P = 0.01) and with both disease recurrence (P = 0.045) and death from disease (P = 0.021). Plasma endostatin above a cut-off point of 70 ng/mL could predict tumour recurrence with a relative risk of 4.7 (95% CI: 1.1-19.7). These data suggest that plasma endostatin and sANG have potential roles as prognostic factors and require further investigation. PMID- 11903370 TI - Effect of UPPP with respect to site of pharyngeal obstruction in sleep apnoea: follow-up at 18 months by overnight recording of airway pressure and flow. AB - Continuous recording of upper airway pressure and airflow can identify the sites of obstructive events during sleep, and their relative distribution along the upper airway segments. A separation of transpalatal and subpalatal obstructive events has been used. The purpose of the present study was to investigate if uvulopalatopharyngoplasty (UPPP) had reduced transpalatal more than subpalatal obstructive events 18 months postoperatively, and to investigate any influence of age and obesity. Fourteen consecutive male patients were investigated by overnight recording of upper airway pressure and airflow before, and 18 months after, UPPP. The localization of obstructive events pre- and postoperatively revealed that the proportion of obstructive events located transpalatally were reduced from 65.1 to 30.5% of all (P < 0.05). Viewing the two sites separately, the reduction in number of transpalatal events (81%) was higher than the reduction in subpalatal events (42%) (P < 0.05). Age + Body Mass Index (BMI) correlated inversely with relative reduction in subpalatal events but did not correlate to any change in transpalatal events. Treatment response with regards to reduction in recorded Apnoea Hypopnoea Index (AHI) was achieved in 9 out of the 11 patients who had mainly transpalatal obstructive events preoperatively. PMID- 11903371 TI - Is shoulder impingement syndrome a problem in otolaryngologists? AB - Shoulder problems are well-documented as an occupational illness. The incidence of occupational shoulder problems is increasing. A postal questionnaire survey was conducted to see if otolaryngologists are more susceptible to shoulder impingement syndrome because of their occupation. Endocrinologists were used as the control group. Among 556 questionnaires sent to otolaryngologists, 367 (64.6%) responses were returned compared with 210 questionnaires sent to endocrinologists, of which 138 (65.7%) responses were returned. A total of 88 (24.0%) of the otolaryngologists had suffered from impingement syndrome compared with 15 (10.9%) of the endocrinologists, which was significantly different. Of those with impingement syndrome, more endocrinologists gave a history of injury or overuse compared with otolaryngologists. There must be another factor, leading to increased incidence of impingement syndrome, which may be because of the continuous flexion, and abduction of their shoulders during examination and operating on patients. PMID- 11903372 TI - Does flying after myringoplasty affect graft take rates? AB - Flying at altitude soon after myringoplasty may theoretically have an adverse effect on graft take rates owing to variation in external air pressure, together with suboptimal Eustachian tube function, causing graft displacement. We wished to assess the effect of flying after myringoplasty by comparing success rates of patients flying within a week postoperatively (n = 37) with a control group (n = 37) by carrying out a retrospective, controlled cohort study. The primary outcome measure was graft success, defined as a 100% closure at first outpatient follow up (2.5 months). Our results showed that there was no difference in early graft success rates between the flying group (32/37: 86%) and non-flying group (29/37: 78%)(P = 0.32; 95% CI of difference: -9% to 27%). In conclusion, flying at altitude in a pressurized environment within a week of myringoplasty does not adversely affect early operative success. PMID- 11903373 TI - Therapeutic improvement of Eustachian tube function: a review. AB - An impaired Eustachian tube is assumed to be an important factor in the pathogenesis of different middle ear diseases. Therefore, several investigators have studied different treatment strategies to improve Eustachian tube function. The aim of this review is to provide a comprehensive summary of the results of these studies on improvement of tubal function. The English language literature was searched systematically to identify all articles that described the effect of different interventions on Eustachian tube function. Although the results were not uniform throughout the different studies and despite several restrictions of the reviewed studies, the results of this review indicate that the function may be improved by medical intervention. However, it seems premature to recommend any of the interventions reviewed in this paper to improve function in humans. More studies, preferably randomized, placebo-controlled trials, should be conducted to assess the efficacy of different interventions. PMID- 11903374 TI - Temporomandibular joint dysfunction following tonsillectomy. AB - We report a prospective, controlled trial to assess temporomandibular joint (TMJ) dysfunction following the use of a Boyle-Davis mouth gag during tonsillectomy. TMJ function was evaluated in patients undergoing tonsillectomy and a control group undergoing nasal surgery preoperatively and 6 weeks postoperatively. The main outcome measures were symptoms and signs of TMJ dysfunction and interincisal distance. A mean reduction of 0.89 mm in interincisal distance (P < 0.01) was noted postoperatively in the tonsillectomy patients. There was no statistically significant reduction of interincisal distance in patients undergoing nasal surgery. There was a statistically significant reduction in interincisal distance in the post-tonsillectomy patients, caused by fibrous healing of the tonsillar bed or fibrous ankylosis of the TMJ. PMID- 11903375 TI - HLA associations with nasopharyngeal carcinoma in Southern Chinese: a meta analysis. AB - The literature relating to human leucocyte antigens (HLA) and nasopharyngeal carcinoma (NPC) identifies conflicting ranges of possible allelic associations. We aimed to clarify this by conducting a systematic review to identify and quantify associations present across all of the available studies. A literature search was performed and, subsequently, a meta-analysis was performed on 13 published studies using both fixed-effects and random-effects models when appropriate. Evidence for positive associations between NPC and the HLA alleles A2, B14 and B46 (P = 1.57 x 10-5, 1.13 x 10-3 and 6.38 x 10-5 respectively) were found, and negative associations were identified for the alleles A11, B13 and B22 (P = 5.42 x 10-3, 0.017 and 0.009). Whereas an association between HLA-B13 or B22 and NPC has not been noted previously, the results for HLA-A2, A11, B14 and B46 are in accordance with published studies. There is evidence that specific allele subtypes or combinations of alleles may carry particular risk for NPC. PMID- 11903376 TI - Radiation-induced cancers of the pharynx and larynx: a study of five clinical cases. AB - Radiation-induced cancer, a rare clinical entity, is often difficult to diagnose and manage. This study reports a series of five cases of radiocarcinogenesis of the pharynx and/or larynx that developed after external radiotherapy. The primary lesion was diagnosed at a mean age of 50 years (+/-12.9) and the radiation induced cancer at a mean age of 59 years (+/-13.1), giving a latent period of 9 years (+/-3.7). Analysis of gammagraphic records indicated that four of the patients had developed a secondary tumour in the penumbra of irradiation fields. In these zones, the delivered dose was between 20 and 80% of the prescribed dose, corresponding to an estimated cumulative mean dose of 14.1-56.3 Gy. These results are compared with data in the literature to determine the diagnostic criteria for radiation-induced cancer, possible predisposition (genetic or acquired) and the dose effect. PMID- 11903377 TI - Surgery for acquired atresia of the external ear canal. PMID- 11903378 TI - Foreword. PMID- 11903379 TI - The influence of age, gender and ethnicity on cadaveric organ recovery rate. AB - In view of the influence of donor factors such as age on graft outcome and the performance standards that measure OPO productivity by the number of organs recovered and transplanted, it is important to understand the relationship of certain donor factors on organ recovery for transplantation from cadaveric donors. We examined the influence of donor age, gender and ethnicity on the number and type of transplanted organs recovered from 598 consecutive cadaveric donors in our OPO between 1994 and July 1999. The highest number of organs/donor ocurs in the 11-20 donor age range and declines significantly with each age range. The type of organ recovered is also influenced by age, but the least effect is on liver recovery. No difference was seen in the number of organs recovered/donor by race. When the data were re-analyzed with regard to renal and extra-renal organs transplanted/million donor population, 78% of the kidneys (n=781/1006) were from the 11-50 age range and 81% of the extra-renal organs (n=822/1,192) were from that age range. Stepwise regression yielded a model where donor age significantly influenced (P=0.001) the number of organs recovered. Finally, the incidence of recovered and transplanted organs was significantly higher in males compared with females for hearts [51% (187/360) vs. 40% (86/214); P<0.006] and pancreata [18% (66/360) vs. 11% (24/214); P<0.02]. The number of organs recovered and transplanted from cadaveric organ donors is influenced predominantly by the age of the donor, with the exception being liver donors. Increasing organ recovery and transplantation of organs from donors from the two age extremes results in less gain in the number of organs/million population than recovery from the 11-50 age range. PMID- 11903380 TI - Presentation and outcomes for organ donation in patients with cerebral gunshot wounds. AB - This study was undertaken to examine the presentation and outcomes relative to solid organ donation in patients with fatal cerebral gunshot wounds at a level I trauma center over a 7-year period. A retrospective chart review of patients with such wounds over the years 1993-99 was completed. Eighty (80) patients were considered potential solid organ donors. Of these, 28 (35%) became organ donors, yielding 97 transplantable organs. Ninety-six percent presented with a GCS of less than 6. Mean SBP on presentation was 130, ranging from 48 to 225. Median time from presentation to death was 18 hours. Intravenous fluids given over the first 6 hours averaged 4.3 liters. Pressors were required in 68% of cases, blood products in 34%. Consent rate for donation was 32% when requested by a physician and 59% when requested by an organ procurement organization (OPO) co-ordinator. No request was made in 15 cases. Patients with fatal cerebral gunshot wounds, but with solid organ donor potential, have a characteristic presentation. Those with hemodynamic stability and those whose hypotension responds promptly to treatment can be expected to have a donor potential despite their devastating brain injury. Minimal time and resources are required to support such patients. Additional organs may have been obtained if the request for donation was consistently separated from the families' notification of brain death, and if the request was initiated by an OPO coordinator rather than a physician. Further, all patients admitted with cerebral gunshot wounds and poor neurologic function should have local OPO referral, potential survival notwithstanding. PMID- 11903381 TI - Organ procurement organization (OPO), best practices. AB - There are currently 59 organ procurement organizations (OPOs) in the United States which serve their assigned geographic areas with variable productivity. Knowledge of organizational characteristics, programs and practices of more successful OPOs may be useful to increase the productivity of less successful OPOs. A preliminary survey of all OPO executive directors in the United States ascertained the most important beneficial and detrimental factors affecting their success. Site visits were then conducted at OPOs based on a selection process utilizing population size, geographic location, minority population, donors per million population and donors per thousand deaths among potential donors. All OPOs were categorized and the highest ranking OPOs in each of seven categories, based on 4 years of national data, were selected for the site visits. Regression analysis and correlation analysis using Pearson's product-moment correlation were performed. The survey to identify the important factors was returned by 47 (77%) of 61 OPOs existent in 1999. The most important beneficial factors identified by responding OPOs were adequate staffing and experience, allocation of responsibilities, hospital development and leadership. The most important detrimental factors were inadequate staffing and experience, poor donor hospital/transplant center/ OPO relationships and failure in the consent process. Site visits of the highest-ranking OPOs demonstrated all had respected, experienced leadership focused on the donation process; efficient mechanisms for resolving allocation or transplant center conflicts; systems for monitoring activity and tracking outcomes; excellent communication between OPO and transplant centers; open internal communication at all levels of the OPO; immediate, on-site response to vascular donor referrals; and volunteer support of public and/or professional education. Regression and correlation analysis demonstrated that as minority population increases, OPO performance declines (P < 0.03). Moreover, independent OPOs were associated with poorer performance regardless of minority population (P < 0.05). All of the successful OPOs visited had strong leadership, excellent donor hospital and transplant center relationships, well-developed communication and innovative methods to deal with their minority populations. Application of these practices within all OPOs could significantly enhance organ donation. PMID- 11903382 TI - HLA phenotypes of ESRD patients are risk factors in the panel-reactive antibody (PRA) response. AB - To determine whether recipient HLA phenotypes are correlated with an increased or decreased risk of alloantibody sensitization in end-stage renal disease (ESRD) candidates for first or repeat kidney transplantation; we analyzed 19440 kidney allograft recipients consisting of 13,216 Caucasians and 6224 non-Caucasians transplanted between 10/87 and 11/98 at South-Eastern Organ Procurement Foundation (SEOPF) member institutions. Relative risk values and 95% confidence limits were obtained using Wolfe's method. Logistic regression was used to adjust for covariates that influence sensitization, i.e. ethnicity, gender, age, pregnancies, transfusions, primary/repeat transplant and living versus cadaver donor. Univariate analysis of the entire cohort indicated that nine HLA allelotypes (DR1,4,7; B8,12,40; A1,2,11) were associated with a significantly reduced risk of sensitization, and five allelotypes (B42,B53; A 10,19,36) were associated with an increased risk of PRA responses. Corrected for the number of statistical comparisons, recipients with DR1, DR4, A1 or A2 were 15% less likely to be sensitized per allelotype. Recipients with B42, B53 or A36 were at increased risk of preformed antibodies, after correction of the P value, for an average of 38% increased risk per allelotype. In the multivariate analysis, HLA phenotypes identified as independent risk factors associated with protection against sensitization were DR1,4,7; B12(44,45); and A1,2, with an average reduced risk of 9% per allelotype. The only independent susceptibility allelotype was A36 with an increased risk of 29%. The A10 (25,26,34,66) group reached borderline significance. We also looked for HLA-DR,-B,-A combinations that could potentially represent protective or at risk haplotypes/genotypes. Stepwise logistic regression identified five combinations associated with protection: DR1-B35-A3; DR1-B35-A2; DR1-B44-A2; DR4-B44-A2; DR7-B57-A1 (RR range 0.83-0.63) with 27% average reduced risk per combination. Phenotype combinations associated with an increased risk of sensitization were: DR2-B44-A2; DR2B53-A2; DR3-B8-A1: DR3-B42 A30; DR6-B42-A30; DR11-B53-A30 (RR range 2.76-1.48) with an average increased risk of 70% per combination. This study provides strong evidence that HLA-linked genes influence the anti-HLA PRA response. The magnitude of the altered PRA response risk in DR-B-A combinations was approximately twice that of the allelotypes at individual loci. HLA-DR genes seemed to contribute most of the altered risk. The correlations between DR types and PRA responsiveness are consistent with the DR types previously regarded as predictors of kidney graft survival. The magnitude of increased PRA risk attributable to an allelotype or combination was approximately twice that associated with a decreased risk. We conclude that some HLA class II-linked genes modulate the PRA response in a clinically significant manner. This immune response gene (Ir) regulation probably operates through polymorphic HLA molecules in their physiologic roles of antigen processing and presentation to helper T cells. PMID- 11903383 TI - IgM antibodies identified by a DTT-ameliorated positive crossmatch do not influence renal graft outcome but the strength of the IgM lymphocytotoxicity is associated with DR phenotype. AB - A positive crossmatch that is rendered negative by treating the serum with the IgM-reducing agent dithiothreitol (DTT) is generally reported not to influence short-term renal graft outcome. Its effect on long-term (> or = 3 years) cadaveric and live-donor transplant function, however, is less clear. We evaluated the effect of IgM antibodies in a DTT-ameliorated positive crossmatch (DTT-APXM) on long-term renal graft outcome in 1,290 consecutive cadaveric renal transplants (8-year survival) and 384 live-donor renal transplants (7-year survival) from patients transplanted between 1990 and 1999. The data show that 1- and 8-year graft survival for cadaveric renal transplants in patients with IgM antibodies (n=72) (DWFG censored = 91% and 65%; DWFG not censored = 90% and 60%) was not significantly different from the group without IgM antibodies (n = 1,218) (DWFG censored = 92% and 71%; DWFG not censored = 87% and 55%) (log-rank = 0.25 for DWFG censored, log-rank = 0.92 for DWFG not censored). The one- and seven year graft survival for live-donor renal transplants in patients with IgM antibodies seen in a DTT-APXM (n = 22) (DWFG censored = 95% and 83%; DWFG not censored = 95% and 66%) was not significantly different from the group without IgM antibodies (n = 362) (DWFG censored = 94% and 81%; DWFG not censored = 92% and 73%) (log-rank = 0.61 for DWFG censored, log-rank = 0.89 for DWFG not censored). DR phenotype was found to be associated with the strong (>40% cell death) IgM reactivity in both black and white patients. In white patients, DR2 was more frequently seen with a strong IgM crossmatch (48.2%) than in molecularly typed controls (28.5%) (P < 0.03) and concomitant with that DR increase, DR4 was decreased in white patients (6.8%) compared with controls (25.5%) (P < 0.02). In black patients with strong IgM reactivity, DR6 was increased in patients (46.1%) compared with controls (20.5%) (P = 0.07) and concomitant with that DR6 increase, DR5 was decreased in frequency in black patients (7.6%) compared with controls (41%) (P < 0.03). These data show that long-term graft survival in renal transplantation is not negatively influenced by the presence of donor-reactive lymphocytotoxic antibodies in the crossmatch ameliorated by serum DTT treatment. They also suggest that the strength of the IgM antibody response is regulated in part by certain gene (s) of the DR region. PMID- 11903384 TI - Positive effect on T-cell regulatory apoptosis by mycophenolate mofetil. AB - The regulatory benefit of apoptosis (activation-induced cell death, AICD) in T cells may be impacted by immunosuppressive agents. We examined this for mycophenolate mofetil (MMF) compared with cyclosporine (CYA). Peripheral blood leukocytes (PBL) were stimulated by either Staph enterotoxin B (SEB) or by anti CD3 plus anti-CD28. Cell division analysis (sequential reduction in carboxyflourescein diacetate succinimidyl ester, CFSE) was used to measure proliferation and determine status of different cell generations. Apoptosis was measured by annexin V staining, and FasL expression by anti-FasL antibody staining, of activated cells using flow cytometry. CSA and mycophenolic acid (MPA, the active agent of MMF) were added in titration in 3-day cultures. We found that CSA caused diminution in apoptosis but MPA increased it with SEB stimulation. The CSA effect on apoptosis was present when a more calcineurin dependent stimulus. anti-CD3+ anti-CD28, was used but the MPA effect was less, producing a decrease only in the undivided cells. To look more directly at the differential effect on calcineurin-dependent AICD gene induction of the two agents, we measured Fas-L expression with anti-CD-3 + CD28 stimulation, and confirmed that CYA caused a major decrement in appearance of Fas-L, whereas MPA caused a converse accumulation of it. This seems to be explained by the block more distal in cell activation, resulting in a build-up of a precursor in the activation pathways. We conclude that MMF treatment may be rationale as an adjunct to calcineurin inhibitor treatment because of its converse effect on T cell regulatory apoptosis. PMID- 11903385 TI - Lifestyle behaviors affect cardiovascular risk status in men 1 year after kidney transplantation. AB - Persons with end-stage renal disease have an accelerated risk for cardiovascular (CV) morbidity and mortality. Unfortunately, their accelerated CV risk persists even after kidney transplantation associated with pretransplant and post transplant vascular disease. number of rejections treated with high-dose steroids, prolonged use of immunosuppressive therapy post-transplant, effects of comorbid chronic conditions and male gender. Unhealthy, modifiable lifestyle practices often augment their CV risk. The purpose of this study was to examine health-related lifestyle behaviors and estimate CV risks of men 1 year following kidney transplantation. Using the Healthier People Network Health Risk Appraisal (HPN-HRA), we examined CV risk characteristics of 34 men (M age=47.3 years) enrolled in an urban, mid-southern transplant center. Participants were assessed via self-administered paper-and-pencil questionnaires and medical record review provided biological data for analyses. HPN-HRA software was used to analyze biological and behavioral characteristics to compute CV risk estimates. Data were analyzed using descriptive, inferential and correlational analyses. The majority of participants reported smoke cessation (n= 19) or never having smoked (n = 8), and consumption of one or less alcoholic drinks per week (n=28). On the other hand, increased risk for heart attack and stroke were associated with advancing age (P<0.0001), white race (P<0.05) and higher systolic blood pressure (P<0.01). While risk for heart attack was associated with current cigarette smoking (P<0.01), risk for stroke was not. Conversely, risk for stroke was associated with higher BMI (P<0.05), risk for heart attack was not. Thus, the study provides evidence that male kidney transplant survivors exhibit a significant number of non-modifiable and modifiable characteristics that contribute to their posttransplant CV risk. PMID- 11903386 TI - Immunosuppressant side effect profile does not differ between organ transplant types. AB - Transplantation has enhanced the quality of life of all transplant recipients, but concern remains regarding the side effects of immunosuppressant drugs. In order to respond to these concerns, a survey to ascertain the side effect profile of transplant recipients was undertaken to identify the impact of chronic immunosuppression on quality of life. A nationwide survey of solid organ transplant recipients was carried out using a newly developed immunosuppressant side effect survey. Kidney, kidney-pancreas, liver and heart recipients responded to the survey (n = 505) and reflect the national distribution based on the UNOS data for organ type, recipient race and gender. The survey had four subscales: emotional burden, life/role responsibilities, mobility and GI distress. A fifth subscale included miscellaneous side effects that are more prevalent during the first 2 years post-transplant. Frequency and severity of each side effect were coded on a scale of 0-4 from 'no problem' to 'always' a problem. The entire range of possible scores (0-160) was reported, reflecting adequate variability in the responses. The sample consisted of 51% males, 77% Caucasians, 15% African Americans, with the remaining 8% other races. There were 225 (44.5%) kidney, 147 (29.1%) liver, 101 (20%) heart and 32 (6.4%) pancreas included. Age ranged from 18-71 years with time since transplant 1-21 years. Overall frequency (12.1 +/- 6.08), severity (10.5 +/- 6.96) and weighted scores (25.4 +/- 19.9) were low suggesting that, as a whole, immunosuppressant side effects, while present, were not severe or troublesome for most patients. Side effect profiles appeared similar among organ types. Differences were detected in the GI distress subscale with the heart recipients reporting significantly less GI distress than liver recipients (13.8 vs. 19.2; P<0.05). Side effect impact on mobility tended to increase between time eras; however, no statistical significance was detected. Side effects are a concern among health-care professionals; however, based on the results of this study, immunosuppressant-related side effects are not detrimental to quality of life and show no differences between types of organ transplanted. PMID- 11903387 TI - Diurnal cyclosporine dosing optimizes exposure and reduces the risk of acute rejection after kidney transplantation. AB - Acute rejection (AR) following transplantation may be due to episodic subtherapeutic cyclosporine (CsA) levels related to diurnal variation of hepatic drug metabolism. We postulated that asymmetrical dosing of CsA based on individualized pharmacokinetic profiles would optimize drug exposure and decrease the risk of AR. We prospectively treated all patients undergoing kidney transplantation with a diurnally split dose of CsA microemulsion given q 12 hours (3.5 mg/kg q a.m., 3.0 mg/kg qPM). Morning doses were adjusted to reach a day time area under the concentration curve (AUC) of 7,800 ng hour/ml (utilizing 2 hour and 6 hour levels) and evening doses were adjusted to a morning trough of 300 ng/ml. Patients received high-dose steroids tapered to 20 mg prednisone by day 6. CsA was started within 36 hours and mycophenolate mofetil (1000 mg q 12 hour) was added on day 3 in most patients and continued for 3 months. Only one patient received antibody induction. Thirty kidneys (67% cadaveric) were transplanted into 28 adult patients (50% African American, 57% men). Therapeutic targets were reached in all patients prior to discharge and maintained during the study period. At 3 months follow-up, there was not a single episode of documented AR and mean creatinine was 1.5 +/- 0.1 mg/ml. Twelve patients required biopsy for allograft dysfunction; however no histological evidence of AR or CsA-toxicity was identified and the creatinine normalized in each case without altering immunosuppression. Patients continued to require increased CsA doses in the AM compared to the PM (P<0.05) throughout the study to maintain target levels. Diurnal dosing of CsA based on individual pharmacokinetic profiles optimizes CsA exposure and reduces the risk of AR during the first 3 months after transplantation. PMID- 11903388 TI - Transplantation of livers from hbc Ab positive donors into HBc Ab negative recipients: a strategy and preliminary results. AB - Here we describe a strategy for using livers from hepatitis B core antibody (anti HBc) positive donors in anti-HBc negative recipients and report our preliminary results. Adult anti-HBc negative recipients were immunized against hepatitis B virus (HBV) prior to transplantation. Liver biopsies from anti-HBc positive, HBs Ag negative donors were performed at the time of procurement to rule out acute hepatitis or chronic liver disease. Donor serum and liver samples were collected for HBV DNA analysis by PCR. Recipients were given HBIG (10000 units, i.v.) during the anhepatic phase of transplantation. Patients were treated with lamivudine (150 mg) beginning on postoperative day (POD) 1. If HBV DNA was not detected in either donor liver or serum by PCR, recipient antiviral therapy was stopped. If donor liver and serum were positive for HBV DNA by PCR, the recipient was maintained on combination lamivudine and HBIG therapy. If HBV DNA was detected in donor liver but not in donor serum, the patient was managed on lamivudine therapy alone. Between February 1999 and June 2000, six anti-HBc negative recipients received liver transplants from anti-HBc positive donors. PCR analysis of serum from the six donors was negative for HBV DNA in each, while donor liver PCR analysis was positive in five of six for HBV DNA. Accordingly, all patients were given HBIG in the anhepatic phase of transplantation and five of six were maintained on daily lamivudine therapy. Follow-up periods have ranged from 2 to 18 months. There has been no emergence of de novo hepatitis B. Serial serum HBs Ag and HBV DNA assays have all proven negative. Moreover, while on lamivudine therapy, 2 patients now have undetectable HBV DNA in hepatic allograft biopsies by PCR analysis. Our strategy for using livers from anti-HBc donors has yielded promising initial results. De novo hepatitis B has not occurred and our data suggest residual hepatitis B virus may be eradicated in recipients maintained on lamivudine therapy. PMID- 11903389 TI - Further improvements in laparoscopic donor nephrectomy: decreased pain and accelerated recovery. AB - Fear of postoperative pain is a disincentive to living donor kidney transplantation. Laparoscopic donor nephrectomy (LDN) was developed in part to dispel this disincentive. The dramatic increase in the number of laparoscopic donor nephrectomies performed at our institution has been in part due to the reduction in postoperative pain as compared to traditional, open donor nephrectomy. We sought to further diminish the pain associated with this surgical technique. The purpose of this study was to compare the efficacy of three different postoperative pain management regimens after LDN. All living kidney donors performed laparoscopically (n=43) between September 1998 and April 2000 were included for analysis. Primary endpoints included postoperative narcotic requirements and length of stay. Narcotic usage was converted to morphine equivalents (ME) for comparison purposes. Patients received one of three pain control regimens (group 1: oral and intravenous narcotics; group II: oral and intravenous narcotics and the On-Q pump delivering a continuous infusion of subfascial bupivicaine 0.5%; and group III: oral and intravenous narcotics and subfascial bupivicaine 0.5% injection). Postoperative intravenous and oral narcotic use as measured in morphine equivalents was significantly less in group III versus groups I and II (group III: 28.7 ME versus group I: 40.2 ME, group II: 44.8 ME; P<0.05). Postoperative length of stay was also shorter for group III (1.8 days) versus group I (2.5 days) and group II (2.9 days). LDN has been shown to be a viable alternative to traditional open donor nephrectomy for living kidney donation. We observed that the use of combined oral and intravenous narcotics alone is associated with greater postoperative narcotic use and increased length of stay compared to either a combined oral and intravenous narcotics plus continuous or single injection subfascial administration of bupivicaine. The progressive modification of our analgesic regimen has resulted in decreased postoperative oral and intravenous narcotic use and a reduction in the length of stay. We recommend subfascial infiltration with bupivicaine to the three laparoscopic sites and the pfannenstiel incision at the conclusion of the procedure to reduce postoperative pain. We believe this improvement in postoperative pain management will continue to make LDN even more appealing to the potential living kidney donor. PMID- 11903390 TI - Multiple renal arteries do not pose an impediment to the routine use of laparoscopic donor nephrectomy. AB - Since the first description by Ratner and collegues in 1996, laparoscopic live donor nephrectomy is gaining wide acceptance in an attempt to minimize the donor morbidity, length of hospital stay and length of time to return to work. It is unknown whether multiple renal arteries pose additional problems with laparoscopic donor nephrectomy. In November 1998, our institution initiated laparoscopic donor nephrectomy program. In the ensuing 19 months, we performed 25 living donor renal transplants, 24 of them using laparoscopic donor nephrectomy. The left kidney was procured in all cases. Eight donor candidates (33%) had two or more renal arteries (two arteries in five patients and three patients). RESULTS: In six cases (25%), findings at surgery differed from the CT angography results (in four cases, CT angiogram reported fewer arteries than were found at surgery and in two cases it reported more). We found no significant differences in both donor outcomes and recipient, based on the presence or absence of multiple renal arteries. Among donor outcomes, we found equivalent results for donor warm ischemia time total donor operating time, and donor length of stay. For recipient outcomes, we found no significant differences between groups for the incidence of acute tubular necrosis (ATN), graft survival and most recent serum creatinine. In one case, we constructed two arteries into a single conduit on the backtable prior to transplantation. However, in most cases with multiple arteries, we implanted the arteries separately into the recipient external iliac artery. Based on this experience, we do not find the presence of multiple renal arteries to be a barrier to the successful use of kidney grafts procured by laparoscopic donor nephrectomy. PMID- 11903391 TI - End-stage renal disease in liver transplants. AB - Renal dysfunction is one of the most significant problems following orthotopic liver transplantation (OLTx). Since the major risk factor for delayed renal dysfunction following OLTx is presumed to be cyclosporine (CsA) nephrotoxicity, it has been suggested that CsA is the most probably cause of end-stage renal disease (ESRD) in this population of patients. To test this hypothesis the records of OLTx patients in our center who developed ESRD requiring dialysis were reviewed. There were 132 consecutive adult patients with end-stage liver disease (ESLD) who received 146 OLTxs between 1990 and 2000. Five patients (3.4%) developed ESRD requiring dialysis. Four of the five patients developed nephrotic range proteinuria prior to reaching ESRD. Renal biopsy in four patients showed focal segmental glomerulosclerosis, diabetic nephropathy, membranous nephropathy and cyclosporine toxicity. The underlying hepatic and metabolic disease may have played a role in the genesis of glomerular diseases in these OLTx patients. Perhaps if more renal biopsies are performed in OLTx patients with chronic renal failure, we might discover that, although CsA/tacrolimus therapy is a definite risk factor for post-transplantation chronic renal failure, other disease processes may also play a significant role. PMID- 11903392 TI - Contact allergy to fragrances: clinical and experimental investigations of the fragrance mix and its ingredients. PMID- 11903393 TI - Protein kinase C and the development of diabetic vascular complications. AB - Hyperglycemic control in diabetes is key to preventing the development and progression of vascular complications such as retinopathy, nephropathy and neuropathy. Increased activation of the diacylglycerol (DAG)-protein kinase C (PKC) signal transduction pathway has been identified in vascular tissues from diabetic animals, and in vascular cells exposed to elevated glucose. Vascular abnormalities associated with glucose-induced PKC activation leading to increased synthesis of DAG include altered vascular blood flow, extracellular matrix deposition, basement membrane thickening, increased permeability and neovascularization. Preferential activation of the PKCbeta isoform by elevated glucose is reported to occur in a variety of vascular tissues. This has lead to the development of LY333531, a PKCbeta isoform specific inhibitor, which has shown potential in animal models to be an orally effective and nontoxic therapy able to produce significant improvements in diabetic retinopathy, nephropathy, neuropathy and cardiac dysfunction. Additionally, the antioxidant vitamin E has been identified as an inhibitor of the DAG-PKC pathway, and shows promise in reducing vascular complications in animal models of diabetes. Given the overwhelming evidence indicating a role for PKC activation in contributing to the development of diabetic vascular complications, pharmacological therapies that can modulate this pathway, particularly with PKC isoform selectivity, show great promise for treatment of vascular complications, even in the presence of hyperglycemia. PMID- 11903394 TI - When and how to start insulin treatment in gestational diabetes: a UK perspective. AB - Gestational diabetes mellitus, however currently defined, is relatively rare in a UK Caucasian population, but is much more common in other ethnic groups. There is likely soon to be better agreement on diagnostic levels of hyperglycaemia in pregnancy, but there is still considerable reluctance to start insulin therapy. There is now good evidence that insulin administered twice daily during the third trimester to mothers who have even a mild degree of hyperglycaemia will reduce fetal size, and in particular fetal adiposity. In relation to recent concepts of the transgenerational passage of Type 2 diabetes and obesity, further epidemiological investigation is required. Insulin treatment in pregnancy may also prove to have a role in prevention of Type 2 diabetes in the next generation. PMID- 11903395 TI - Normalization of pregnancy outcome in pregestational diabetes through functional insulin treatment and modular out-patient education adapted for pregnancy. AB - AIM: To investigate whether modular out-patient group education for flexible, Functional Insulin Treatment (FIT) adapted for pregnancy can eliminate diabetes associated neonatal complications in pregestational diabetes. RESEARCH DESIGN AND METHODS: Outcome analysis of the modular out-patient group education and FIT based on separate insulin dosages for fasting, eating or correcting hyperglycaemia in 76 consecutive pregnancies (in 20 cases first after conception) of 59 patients with pregestational diabetes (Type 1 diabetes, n = 54). CONTROLS: (a) diabetic pregnancies: historical controls; (b) non-diabetic pregnancies: retrospective case-controlled study; (c) population-based data of all Austrian newborns registered within the respective time period. RESULTS: HbA1c of 113 +/- 18% of mean value (= 100%) of non-diabetic, non-pregnant population (103 +/- 14% during the last pregnancy trimester), and self-monitored blood glucose of 5.6 +/- 0.7 mmol/l (5.3 +/- 0.7 mmol/l during the last trimester) was achieved throughout all FIT pregnancies. Severe hypoglycaemia occurred in 14 pregnancies. The gestational age at delivery was 39.2 +/- 1.5 weeks (four cases (5.4%) < 37 weeks) with a birth weight of 3305 +/- 496 g. Four newborns (5.3%) were above the 90th, and nine (11.8%) below the 10th percentile for weight of reference population based data. Hypoglycaemia was recorded in six newborns (8%). Malformations were found in two infants whose mothers booked for diabetes FIT education only after conception. The caesarean delivery rate was 25%. In comparison with historical diabetic pregnancy controls we demonstrated a reduction in major complications, and compared with non-diabetic women, a lowering of diabetes-related neonatal complication rates to general population levels. CONCLUSIONS: Structured, comprehensive, modular out-patient group education promoting self-choice of insulin dose for flexible, normal eating prior to conception normalizes pregnancy outcome in diabetes. PMID- 11903397 TI - Enhanced endothelial activation in diabetic patients with unstable angina and non Q-wave myocardial infarction. AB - AIMS: Diabetes mellitus (DM) is associated with chronic endothelial dysfunction. Diabetic patients presenting with acute coronary syndromes have a worse prognosis than non-diabetics. An acute inflammatory reaction at the site of coronary plaque rupture and increased expression of surface and soluble cellular adhesion molecules (CAMs) are pathological features of acute coronary syndromes. We set out to characterize the expression of soluble CAMs in patients with and without diabetes presenting with unstable angina (UA) and non Q-wave myocardial infarction (NQMI). METHODS: Patients presenting with UA and NQMI had serum samples taken on presentation, after 72 h and then 3, 6 and 12 months after discharge. Levels of soluble intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin and P-selectin were measured using an ELISA technique. RESULTS: We studied 15 diabetic patients and 15 age- and sex matched non-diabetic patients presenting with either UA or NQMI. Levels of soluble E-selectin were elevated in the diabetic patients in comparison with the non-diabetic patients at all measured time points: 74 +/- 10 ng/ml vs. 47 +/- 3 ng/ml, P < 0.03 at t = 0 h, 55 +/- 5 ng/ml vs. 38 +/- 2 ng/ml, P < 0.02 at t = 72 h. However, levels of soluble P-selectin were lower in the diabetic cohort during follow-up: 134 +/- 15 ng/ml vs. 225 +/- 32 ng/ml, P < 0.02 at t = 3/12 and 112 +/ 8 ng/ml vs. 197 +/- 23 ng/ml, P < 0.02 at t = 6/12. There was no significant difference in levels of soluble ICAM-1 and VCAM-1 between diabetic and non diabetic patients. CONCLUSIONS: Levels of soluble E-selectin are significantly elevated in diabetic patients presenting with UA and NQMI in comparison with non diabetics. This finding may reflect enhanced endothelial activation which may contribute to the adverse prognosis of diabetic patients with acute coronary syndromes. PMID- 11903396 TI - Brain natriuretic peptide increases urinary albumin and alpha-1 microglobulin excretion in Type 1 diabetes mellitus. AB - Atrial natriuretic peptide (ANP) increases urinary albumin excretion in Type 1 diabetes mellitus (DM). Brain natriuretic peptide (BNP) is structurally and functionally related to ANP, but its effect on urine albumin excretion rate (UAER) is unknown. AIMS: To compare the albuminuric effects of intravenous infusion of ANP and BNP, and to assess the effect of both peptides on tubular protein excretion. METHODS: Eight subjects with Type 1 DM were randomised to a three leg, double blind, and placebo controlled study. On each study day, subjects were euglycaemic clamped and subsequently water loaded (20 mL/kg orally, plus urine losses) to steady state diuresis. When in steady state, creatinine clearance was estimated in three separate 1 hour periods. At the end of the first period, a 1 hour intravenous infusion of either placebo, ANP 0.025 microg/kg/min, or BNP 0.025 microg/kg/min was administered. There followed a 1 hour recovery period. Urine was collected at 15 min intervals for estimation of urine albumin (ACR) and alpha1 microglobulin creatinine ratio (MCR). Results were analysed by anova. RESULTS: Creatinine clearance was similar on the three study days, and was unaltered by any infusion. ACR was unaltered by placebo (1.3 +/- 0.5-1.2 +/- 0.4 mg/mmol, mean +/- SD, p = 0.81), but increased compared to placebo with infusion of both ANP (1.2 +/- 0.4-9.8 +/- 8.4 mg/mmol, P = 0.0004), and BNP (1.1 +/- 0.4 13.4 +/- 8.6 mg/mmol, P = 0.0001). The MCR was unaltered by placebo infusion (P = 0.89), but increased compared with placebo after infusion of ANP (5.4 +/- 0.9 12.3 +/- 4.2 mg/mmol, P < 0.0001), and BNP (5.4 +/- 0.8-12.1 +/- 2.5 mg/mmol, P < 0.0001). CONCLUSIONS: Intravenous infusion of BNP and ANP both increase the urine excretion of albumin and the tubular protein alpha1 microglobulin, independent of creatinine clearance. PMID- 11903398 TI - Non-diabetic relatives of Type 2 diabetic families: dietary intake contributes to the increased risk of diabetes. AB - AIMS: Non-diabetic first degree relatives of Type 2 diabetic patients are at increased risk of developing diabetes and cardiovascular disease. This is assumed to reflect a shared genetic predisposition. The aim of this study was to test the hypothesis that lifestyle factors, specifically dietary factors, are also important to the increased risk in non-diabetic relatives. METHODS: Dietary intake was assessed using a validated food frequency questionnaire in 149 non diabetic first degree relatives (age 20-65 years) from families of North European extraction with two or more living Type 2 diabetic family members, and 143 age- and sex-matched control subjects from the background population with no family history of diabetes. RESULTS: Relatives reported higher absolute intakes of total fat (mean (95% confidence intervals) 83 (76-91) vs. 71 (66-76) g/day, P = 0.01), saturated fat (SFA; 39 (36-43) vs. 33 (30-36) g/day, P < 0.01) and cholesterol (391 (354-427) vs. 318 (287-349) mg/day, P < 0.01), and a lower intake of non starch polysaccharide (P < 0.05). Considered as percentage of total daily energy intake, relatives had higher intakes of total fat (P < 0.01) and SFA (P < 0.02), and a lower intake of carbohydrate (P < 0.02). These differences remained after exclusion of suspected under- and over-reporters of dietary intake. CONCLUSIONS: Non-diabetic relatives of Type 2 diabetic patients were found to consume diets that will promote rather than prevent the development of diabetes and cardiovascular disease. This suggests that the increased risk to non-diabetic relatives is therefore not entirely genetic, and there is scope for decreasing the risk through lifestyle modification. PMID- 11903399 TI - Associations between apolipoprotein E phenotype, glucose metabolism and cognitive function in men. An explorative study in a population sample. AB - AIMS: To investigate the associations of the apolipoprotein E phenotype (apoE) and disturbed glucose metabolism with cognitive function in a random population sample. METHODS: A cross-sectional study was conducted, in which 528 men aged 54 or 60 years were recruited randomly from a larger population-based sample of 1516 men. A subject was defined as having abnormal glucose tolerance (AGT), if he had a clinical diagnosis of diabetes, with either dietary or oral antidiabetic treatment or showed impaired glucose tolerance in an oral glucose tolerance test. The subjects were divided into three groups according to apolipoprotein E phenotypes: (a) E2/4, E3/4 or E4/4 (apoE E4); (b) E 3/3 (apoE E3); and (c) E2/2 or E2/3 (apoE E2). Memory function was examined using a word-list learning with Buschke's selective reminding method and test. Executive functions were assessed with the Trail Making Test A and B. RESULTS: Those subjects with apoE E2 and abnormal glucose metabolism demonstrated the worst cognitive executive control compared to other groups. Simple cognitive speed did not differ between the groups. CONCLUSIONS: The exploratory analyses revealed that subjects with apoE E2 allele and AGT had worse glycaemic control and cognitive executive control compared to other groups. Different apolipoprotein phenotypes together with impaired glucose tolerance may have different cumulative adverse effects on age related cognitive performance. Some subgroups of subjects may be especially vulnerable to cognitive impairment. PMID- 11903400 TI - Population-based survey and analysis of trends in the prevalence of diabetic nephropathy in Type 1 diabetes. AB - AIMS: To determine the prevalence of the various stages of diabetic nephropathy in Type 1 diabetes in a population-based survey. To make direct comparison with results from previous published studies to assess current trends. METHODS: Identification of all Type 1 patients using a population-based diabetes register. Urine samples for albumin assay were obtained at clinic visit and by postal request. Prevalence rates were calculated specifically for direct comparisons with previously published surveys. RESULTS: This study and European data from the 1990s show a clear reduction in the cumulative prevalence of microalbuminuria and established nephropathy compared with surveys from Copenhagen (1985), Pittsburgh (1986-8) and Boston (1992). In North Wales in 1999 the overall cumulative prevalence of microalbuminuria was 27.2% (95% confidence interval (CI) 24.3 30.1%) and established nephropathy was 9.6% (7.7-11.5%). Comparisons with data from EURODIAB and from Spain indicated similar results, although the prevalence of microalbuminuria was lower in North Wales than in the EURODIAB study. Significantly lower rates of nephropathy were seen in more recent Swedish cohorts. Diabetic nephropathy remains more common in males. Microalbuminuria before 10 years duration of diabetes was seen at all post-pubertal ages. CONCLUSIONS: Rates of nephropathy in Europe in the 1990s are lower than a decade ago. Modern methods of management are therefore associated with demonstrable benefit at the population level. The lowest rates of nephropathy are associated with optimum glycaemic control in Swedish data, indicating the importance of metabolic in addition to haemodynamic factors. PMID- 11903401 TI - Use of oral fluorescein angiography in the diagnosis of macular oedema within a diabetic retinopathy screening programme. AB - AIMS: To assess if oral fluorescein angiography (OFA) is a suitable screening method to detect macular oedema in diabetic retinopathy. METHODS: Eighty-four diabetic patients were included in the study. They were from a consecutive series of patients attending the diabetic eye-screening clinic, with retinopathy at the macula requiring ophthalmology assessment. All patients were subsequently examined in the eye hospital, by ophthalmologist slit lamp biomicroscopy assessment as the gold standard, followed by oral fluorescein angiography. RESULTS: This study indicates a sensitivity of 92% and specificity of 81%. Only 4.8% of patients developed a minor reaction to oral fluorescein; 84.5% of images were of good quality. CONCLUSIONS: Oral fluorescein angiography is an efficient and highly sensitive tool for the detection of macular oedema. It can be used as an adjunct in the diabetic screening service to identify patients with oedema within a disc diameter of the macula. Ultimately it will ensure that only necessary and smaller numbers of patients are referred to ophthalmologists. PMID- 11903402 TI - Biochemical marker for cerebral oedema complicating severe diabetic ketoacidosis. PMID- 11903404 TI - Hyperhomocysteinaemia and neuropathy in Type 2 diabetes. PMID- 11903403 TI - Serum S-100beta protein as a biochemical marker for cerebral oedema complicating severe diabetic ketoacidosis. PMID- 11903405 TI - Hypoglycaemia induced by disopyramide in a patient with Type 2 diabetes mellitus. PMID- 11903406 TI - A possible hypoglycaemic effect of maitake mushroom on Type 2 diabetic patients. PMID- 11903409 TI - Hepatotoxicity of thiazolidinediones. PMID- 11903410 TI - Insulin release and suppression by tacrolimus, rapamycin and cyclosporin A are through regulation of the ATP-sensitive potassium channel. AB - AIM: By focusing on the pancreatic beta cell response to tacrolimus, cyclosporin A (CsA) and rapamycin we hoped to identify immunophilin, calcineurin and/or novel mechanism involvement and advance the understanding of immunosuppressant regulated insulin control. METHODS: A glucose responsive beta cell model was established in which the glucose response was blocked by immunosuppressant treatment and this model was used to further characterise this effect. Quantification of insulin release to immunosuppressants and specific inhibitors was used to identify the mechanism involved. RESULTS: It was found that upon the addition of tacrolimus, rapamycin, or CsA, rapid and significant exocytosis of cellular insulin was seen. A dose response study of this effect revealed optimal concentration windows of 50- 80 nm for tacrolimus, 100-300 nm for rapamycin, and 7-12 mm for CsA in RIN-5F cells. Optimal insulin release for HIT-T15 cells was similar. Additional experiments demonstrate that immunosuppressant pretreatment blocked the subsequent immunosuppressant induced insulin release but not that of a thapsigargin control, suggesting that suppression and release are non-toxic, specific and in the same pathway. Further experiments showed that this insulin release was a calcium dependent process, which was blocked by inhibitors of l type calcium channels. Continued studies showed that the specific ATP-sensitive potassium channel agonist diazoxide (150 mm) also blocked immunosuppressant induced insulin release. CONCLUSIONS: A model that fits this data is a novel calcineurin-independent immunophilin mediated partial closing of the ATP sensitive potassium channel, which would lead to an initial insulin release but would reduce subsequent responses through this pathway. PMID- 11903411 TI - Effects and serum levels of glibenclamide and its active metabolites in patients with type 2 diabetes. AB - OBJECTIVE: To study the effects and serum levels of glibenclamide (Gb) and its active metabolites in patients on chronic Gb medication on different daily doses. MATERIAL AND METHODS: Fifty patients with type 2 diabetes on regular Gb therapy (1.75-14.0 mg daily). Blood samples were taken immediately before and 90 min after regular Gb intake. A standardized breakfast was served 30 min after drug intake. Serum insulin and proinsulin levels were determined by ELISA methods without cross-reactivities. Serum drug levels were determined by HPLC. Fischer's R to Z-test (correlation coefficients) and paired Student t-tests were used when comparing values within the entire group and unpaired non-parametric Mann-Whitney tests were used when comparing high and low dose levels. A p-value < 0.05 was considered significant. RESULTS: There were significant correlations between daily Gb dose, on the one hand, and, on the other, HbAlc (r = 0.55), Delta insulin (r = - 0.59) and Delta-proinsulin (r = - 0.52) levels. Significant correlations between Gb therapy duration and insulin (r = - 0.40) and proinsulin (r = - 0.34) secretion and between Gb dose and ratio proinsulin/insulin (RPI) at both time points (r = 0.32 and 0.30) were also found. The RPI was lower after Gb intake. In patients on > or = 10.5 mg steady state serum metabolite levels (Ml and Ml + M2) were higher (29(0-120) and 33 (0-120) ng/ml) than those of Gb itself (18(0-64) ng/ml). A great inter-subject variability in Gb levels at both time points was seen. CONCLUSIONS: Our results indicate that, in patients on chronic medication, Gb is capable of stimulating both insulin and proinsulin secretion; the effect on insulin release is relatively greater. The effect was more pronounced in patients on a low Gb dose, either because of less impaired beta cells in those receiving low doses, or due to reduced sulphonylurea sensitivity in those on high dosage (down-regulation). In patients on a daily dose of 10.5 mg or more, serum metabolite levels of clinical relevance were demonstrated; the metabolites may contribute to hypoglycaemic events. PMID- 11903412 TI - The effects of intensive glycaemic control on body composition in patients with type 2 diabetes. AB - AIM: To examine the effects of improved glycaemic control over 20 weeks on the type and distribution of weight change in patients with type 2 diabetes who at baseline have poor glycaemic control. METHODS: Forty-three patients with type 2 diabetes and HbA1c > 8.9% were randomised to either intensive glycaemic control (IC) n = 21 or usual glycaemic control (UC) n = 22 for 20 weeks. Dual energy X ray absorptiometry was used to assess the type and distribution of weight change during the study. RESULTS: After 20 weeks HbA1c was significantly lower in patients randomised to IC than UC (HbA1c IC 8.02 +/- 0.25% vs. UC 10.23 +/- 0.23%, p < 0.0001). In the IC group weight increased by 3.2 +/- 0.8 kg after 20 weeks (fat-free mass increased by 1.8 +/- 0.3 kg) compared to 0.02 +/- 0.70 kg in UC (p = 0.003). The gain in total body fat mass comprised trunk fat mass (IC 0.94 +/- 0.5 kg vs. UC 0.04 +/- 0.4 kg, p = 0.18) and peripheral fat mass (total body fat - trunk fat) (IC 0.71 +/- 0.32 kg vs. UC -0.21 +/- 0.28 kg, p = 0.04). Blood pressure and serum lipid concentrations did not change over time in either group. CONCLUSIONS: Intensive glycaemic control was associated with weight gain which was distributed in similar proportions between the central and peripheral regions and consisted of similar proportions of fat and fat-free mass. Blood pressure and serum lipid concentrations were not adversely affected. PMID- 11903413 TI - The effects of magnesium sulphate and EDTA in the hypercholesterolaemic rabbit. AB - Numerous clinical reports suggest the beneficial effects of chelation therapy for the treatment of atherosclerosis. However, the results of these studies are inconclusive and controversial. The purpose of this present study was to examine the prophylactic and therapeutic effects of chelation liquid (CHL) in experimental atherosclerosis. Twenty New Zealand white rabbits were fed a 1% cholesterol-supplemented diet for 45 days. In the prophylactic phase of the study subcutaneous 300 mg EDTA + 500 mg magnesium sulphate (MgSO4) injections (five rabbits) and isotonic saline (five rabbits) were given to test and control groups, respectively, along with cholesterol rich diet. The CHL treatment ameliorated the rise of serum cholesterol and serum triglyceride concentrations, lowered serum calcium concentrations and reduced the aortic atheroma. In the therapeutic phase of the experiment the cholesterol diet was stopped and the remaining 10 animals were returned to normal diet. Five of these rabbits were given CHL injections and other five animals were given isotonic saline injections for 121 days. Although the level of cholesterol and triglyceride were not significantly different in the two groups, the serum calcium concentration and the percentage of the area of flate aortic specimen occupied by atheroma were significantly lower in the CHL treated rabbits as compared to controls. It is concluded that CHL injections have a definite prophylactic effect on atherogenesis in the cholesterol-fed rabbit, and may have some therapeutic value in the regression phase. Further confirmatory studies are suggested. PMID- 11903414 TI - Effect of acarbose on weight maintenance after dietary weight loss in obese subjects. AB - AIMS: Acarbose is a well established antidiabetic drug and is known to exert a modest weight-lowering effect. The aim of this study was to assess the potential of acarbose to improve weight maintenance after a substantial weight loss by dietary measures in obese subjects. DESIGN: Randomised, double-blind, placebo controlled trial of the effect of acarbose on weight change over a 6-month follow up period. PATIENTS AND METHODS: One hundred and ten obese subjects with a BMI > or = 32 and < or = 38 kg/m2 were included in the study and underwent a 10-16-week very-low-calorie diet programme to initiate weight loss. Then, subjects were randomised to receive either acarbose or placebo for 26 +/- 2 weeks. The primary variable was body weight. The primary efficacy analysis was performed in the per protocol population (n = 75). RESULTS: After an initial mean weight loss of 10.0 +/- 3.4 kg, 54 subjects received acarbose at increasing dosage and 56 subjects received placebo treatment. After 14 weeks of follow-up, there was no change in body weight in the two groups. After 26 weeks, completed by 37 subjects in the acarbose group and by 38 subjects in the placebo group, a small weight regain of 0.6 kg was documented in the latter, whereas no weight increase was observed under acarbose treatment (p = 0.38, analysis of covariance with initial body weight as covariable). CONCLUSION: In obese individuals who undergo a hypocaloric diet and achieve a substantial loss of body weight, acarbose treatment provides only a very modest, not significant benefit to stabilise weight reduction. Thus, acarbose is not a useful adjunct to improve weight maintenance in obese subjects after weight loss. PMID- 11903415 TI - Long-term glycaemic improvement after addition of metformin to insulin in insulin treated obese type 2 diabetes patients. AB - AIM: To assess the adjunct effect of metformin to insulin in type 2 diabetes. METHODS: Obese and overweight type 2 diabetes patients treated with insulin for at least 1 year, and with poor glycaemic control (HbA1c > upper reference level + 2%), were included in a randomised, double-blind, placebo-controlled study. Patients were treated for 12 months with either metformin (850 mg b.i.d.) or placebo added to their usual insulin, which was stabilized during a 3-month placebo run-in period, but thereafter attempted to be unchanged. RESULTS: Thirty seven patients were included. Two patients dropped out during run-in. There were no differences between the metformin (n = 16) and placebo (n = 19) group at baseline. Most patients received multiple insulin injections. Metabolic control was improved by addition of metformin. Mean change in HbA1c from baseline showed highly significant difference between groups at 3, 6, 9 and 12 months. Mean change (percentage units +/- s.d.) at 12 months was -1.1 +/- 0.7% vs. + 0.3 +/- 0.8% (p < 0.001) for HbA1c and -1.4 +/- 2.1 mmol/l vs. + 0.6 +/- 2.2 mmol/l (p = 0.025) for fasting blood glucose. Mean low density lipoprotein (LDL) cholesterol change differed slightly at 6 months, but not at 12 months. There were no changes in insulin dose, blood pressure, body weight, triglycerides, total- and high density lipoprotein (HDL) cholesterol, fibrinogen, C-peptide and laboratory safety variables, including serum B12. Combination therapy was well-tolerated with the same adverse event rate as insulin alone, but more patients with diarrhoea. CONCLUSION: Addition of metformin to insulin induced and maintained clinically significant and consistent long-term reduction of hyperglycaemia in obese, insulin-treated type 2 diabetes patients. PMID- 11903416 TI - Effects of chronic murine and human leptin infusion on plasma leptin and corticosterone levels and energy balance in lean Zucker rats. AB - AIM: To clarify whether centrally delivered leptin can access the circulation and to determine to what extent the effects of i.c.v. h-leptin and m-leptin on body weight and plasma corticosterone are due to reduced food intake. METHODS: Male lean Zucker rats were infused i.c.v. with recombinant m-leptin or h-leptin (42 microg/day) for 7 days. Terminal plasma leptin levels were measured using selective r-leptin, m-leptin and h-leptin RIA. Plasma h-leptin and corticosterone levels were determined on days 0, 2, 4 and 6 of h-leptin infusion. Interscapular brown adipose tissue weight and UCP-1 mRNA expression (an indicator of thermogenic capacity) were also measured. RESULTS: The terminal plasma leptin level was elevated (from 2.2 +/- 0.4 to 42.7 +/- 20.2 ng/ml) in the h-leptin treated lean rats to levels similar to those in vehicle i.c.v. infused fa/fa rats (72.2 +/- 4.7 ng/ml), but this was only detectable when the h-leptin radioimmunoabsorbent assay (RIA) was used. Further, both m-leptin and h-leptin infusions in lean rats elevated terminal plasma corticosterone (352 +/- 37 and 389 +/- 55 ng/ml, respectively) to levels similar to those in i.c.v. rats (386 +/ 62 ng/ml), whereas diet-restriction by pair-feeding, with the h-leptin group, in lean rats had no effect (207 +/- 45 ng/ml). The increase in plasma corticosterone level coincided with the maximum hypophagic effects of leptin and preceded the appearance and sustained elevation of exogenous human leptin in the circulation. Both m-leptin and h-leptin i.c.v. infusion reduced body weight gain (3% and 4%, respectively, compared to pair-fed group) and increased UCP-1 expression (11-fold and 16-fold, respectively) in lean rats. However, h-leptin elicited an earlier effect than m-leptin on body weight, manifested as an earlier reduction in food intake and greater increase in UCP-1 expression. h-Leptin also elicited a greater reduction in body weight gain than did pair-feeding. CONCLUSIONS: Intracerebroventricular-infused m-leptin or h-leptin was detected in the circulation. Furthermore, m-leptin and h-leptin elevated plasma corticosterone levels and h-leptin caused some weight loss in lean rats independently of its suppression of food intake. The elevation of corticosterone levels in the lean rats may be a mechanism whereby they resist excessive weight loss in response to leptin. PMID- 11903417 TI - The NEPI antidiabetes study (NANSY). 1: short-term dose-effect relations of glimepiride in subjects with impaired fasting glucose. AB - AIM: NANSY is a randomised, placebo-controlled Swedish-Norwegian study which aims to include 2 x 1112 male and female subjects with impaired fasting glucose (IFG), to assess whether conversion to type 2 diabetes can be delayed by addition of sulphonylurea to dietary regulation and increased exercise. This pilot study was conducted to find the optimum dose of glimepiride in NANSY. METHODS: In a double blind trial in primary care 25 IFG subjects were in random order exposed to single doses and one-week treatments with 0 (placebo), 0.5, 1.0 and 2.0 mg glimepiride once daily. The optimum dose was assessed by measuring blood glucose during oral 75 g glucose tolerance test (OGTT), comparing fasting blood glucose, and the area under the blood glucose curve (AUC), and by monitoring hypoglycaemic events. RESULTS: With single doses, there was a clear dose-response relationship for the reduction in AUC, with a statistically significant difference only between placebo (mean 1981, 95% confidence intervals (CI) 1883-2078) and 2 mg glimepiride (mean 1763, 95% CI 1665-1861). However, following 1-week treatments, the only significant difference was between placebo (mean 1934, 95% CI 1856-2012) and 1 mg glimepiride (mean 1714, 95% CI 1637-1792). Correspondingly, the only statistically significant difference in fasting blood glucose day 7 was between placebo (5.87 mmol/l, 95% CI 5.68-6.05 mmol/l) and 1 mg glimepiride (5.42 mmol/l, 95% CI 5.21-5.62 mmol/l). Chemical hypoglycaemia was common but hypoglycaemic symptoms were rare and similar between the active doses, and easily countered by the subjects. CONCLUSIONS: The sulphonylurea dose-effect curve may be bell shaped, perhaps due to down regulation of sulphonylurea receptors during chronic exposure. Alternatively, the finding could be a rebound phenomenon, secondary to preceding hypoglycaemia. The optimum dose for NANSY was found to be 1 mg glimepiride. PMID- 11903418 TI - Blood glucose- and triglyceride-lowering effect of trans-dehydrocrotonin, a diterpene from Croton cajucara Benth., in rats. AB - AIM: The objective of this study was to assess in rats the antidiabetic effects (i.e. reduction of hyperglycaemia and hypertriglyceridaemia) of trans dehydrocrotonin (t-DCTN), a bioactive diterpene isolated from the popular medicinal plant Croton cajucara. METHODS: Hyperglycaemia was induced by intraperitoneal injection of streptozotocin (STZ) and hypertriglyceridaemia by oral administration of ethanol in rats. The blood glucose levels were measured by the glucose oxidase method using commercially available enzyme kits. RESULTS: Treating rats with t-DCTN (50 mg/kg) significantly reduced STZ-induced increases in blood glucose levels as well as ethanol-induced increases in blood triglycerides. CONCLUSION: The results suggest that t-DCTN has an antidiabetic potential that warrants further research on its mechanism and clinical significance. PMID- 11903419 TI - Insulin-like growth factor I, epidermal growth factor and transforming growth factor beta expression and their association with intrauterine fetal growth retardation, such as development during human pregnancy. AB - AIM: Fetal intrauterine growth retardation (IUGR) is one of the most common obstetric problems, with a frequency of 12% in Mexico. In the past, investigations have focused on extrinsic causes of IUGR. More recent studies have examined the intrinsic factors that cause fetal intrauterine growth. Maintenance of fetal growth has been attributed to insulin-like growth factor (IGF), epidermal growth factor (EGF) and transforming growth factor beta (TGF-beta). The objective of this study was to assess the levels of these growth factors during pregnancy and to determine whether or not low concentrations are associated with IUGR. METHODS: Nine women whose pregnancies were complicated by IUGR and a group of nine women whose pregnancies exhibited normal fetal intrauterine growth were studied. IUGR was determined by sonography and confirmed by weight at birth. Venous blood samples were taken from both groups of pregnant women at the end of each trimester. Enzyme-linked immunosorbent assays, immunoradiometric assays and radioimmunoassays were used to process samples, and the results were analysed by anova. RESULTS: IGF-I levels increased in both groups during pregnancy, but the increase was lower (p < 0.001) in the IUGR group throughout pregnancy and at delivery. EGF did not show any significant changes during pregnancy. Blood TGF beta levels varied only during the first trimester of pregnancy. The differences were not statistically significant. However, TGF-beta concentrations were higher in the pregnancies with IUGR. Women in the IUGR group were smaller than in the control group (p < 0.05), and, using the covariance test (p < 0.05), this was found to be correlated with IGF-I levels but not with EGF or TGF-beta levels. CONCLUSIONS: Changes in fetal weight might be explained by the different concentrations of IGF. The structural homology between IGF-1 and insulin could mean that the presence of higher levels of IGF would result in a increased energetic metabolism that could contribute to fetal growth. EGF levels were not related to IUGR, and TGF-beta levels increased only during the first 3 months in the IUGR group. This observation correlates with the in vitro action of TGF-beta as a negative factor of growth, but as a positive support for sustaining early pregnancy. Our data illustrates that low height represents an increased risk factor for IUGR. These data also correlate with the studies involving extrinsic factors. PMID- 11903421 TI - Treatment of high-risk diabetic patients with angiotensin II receptor blockers. AB - In the United States, approximately 16 million people have diabetes; 90-95% have type 2 diabetes. They are at increased risk of developing hypertension and cardiovascular disease (CVD). The benefits of treating hypertension in diabetic patients and the potential to delay complications and reduce mortality have been demonstrated in clinical trials. Increasing evidence shows that angiotensin converting enzyme (ACE) inhibitors and angiotensin II (Ang II) receptor blockers (ARBs) may be equally effective in delaying progressive renal disease in diabetic patients. Large, multicentre trials are ongoing to confirm the efficacy and superior safety profile of ARBs in this population. PMID- 11903420 TI - Losartan modifies glomerular hyperfiltration and insulin sensitivity in type 1 diabetes. AB - AIM: The effect of the angiotensin II receptor antagonist losartan on renal haemodynamics and insulin-mediated glucose disposal was examined in normotensive, normoalbuminuric type 1 diabetic patients using a double-blind, placebo controlled, cross-over design. METHODS: Diurnal blood pressure, glomerular filtration rate (GFR, determined using [125I]-iothalamate), renal plasma flow (RPF, determined using [131I]-hippuran) and urinary albumin excretion rate (UAE) were measured, and a hyperinsulinaemic, euglycaemic clamp with indirect calorimetry was performed in nine patients (age 30 +/- 7 years (mean +/- s.d.), HbA1c 8.1 +/- 1.1%) following 6 weeks' administration of either losartan 50 mg/day or placebo. RESULTS: Diurnal blood pressure was significantly reduced after losartan compared with placebo (122/70 +/- 11/8 vs. 130/76 +/- 12/6 mmHg, p < 0.05). A significant decline in GFR (133 +/- 23 vs. 140 +/- 22 ml/min, p < 0.05) and filtration fraction (FF; GFR/RPF) (24.6 +/- 3.5 vs. 26.2 +/- 3.6%, p < 0.05) was observed in the losartan vs. placebo groups. RPF and UAE did not change. Isotopically determined glucose disposal rates were similar after losartan and placebo in the basal (2.61 +/- 0.53 vs. 2.98 +/- 0.93 mg/kg/min) and insulin-stimulated states (6.84 +/- 2.52 vs. 6.97 +/- 3.11 mg/kg/min). However, the glucose oxidation rate increased significantly after losartan vs. placebo in the basal state (1.72 +/- 0.34 vs. 1.33 +/- 0.18, mg/kg/min, p < 0.01) and during insulin stimulation (2.89 +/- 0.75 vs. 2.40 +/- 0.62 mg/kg/min, p < 0.03). Basal and insulin-stimulated non-oxidative glucose disposal tended to decrease after losartan; however, this was not significant. Endogenous glucose production and lipid oxidation were unchanged after treatment and similarly suppressed during hyperinsulinaemia. Glycaemic control, total cholesterol, high-density lipoprotein (HDL)-cholesterol and triglycerides were stable in both losartan and placebo groups. CONCLUSIONS: Losartan reduces blood pressure, glomerular hyperfiltration and FF, and improves basal and insulin-stimulated glucose oxidation in normotensive, normoalbuminuric type 1 diabetic patients. PMID- 11903423 TI - Paediatric 'best sellers': a cornucopia of ideas. PMID- 11903424 TI - Publish or perish: easier said than done. PMID- 11903425 TI - Paediatic emergency departments: old needs, new challenges and future opportunities. PMID- 11903426 TI - Current trends in the management of major paediatric trauma. PMID- 11903427 TI - Practical approach to the febrile child in the emergency department. PMID- 11903428 TI - What every emergency physician needs to know about childhood asthma. PMID- 11903429 TI - Analysis of the study design and manuscript deficiencies in research articles submitted to Emergency Medicine. AB - OBJECTIVE: To describe and analyse the study design and manuscript deficiencies in original research articles submitted to Emergency Medicine. METHODS: This was a retrospective, analytical study. Articles were enrolled if the reports of the Section Editor and two reviewers were available. Data were extracted from these reports only. Outcome measures were the mean number and nature of the deficiencies and the mean reviewers' assessment score. RESULTS: Fifty-seven articles were evaluated (28 accepted for publication, 19 rejected, 10 pending revision). The mean (+/- SD) number of deficiencies was 18.1 +/- 6.9, 16.4 +/- 6.5 and 18.4 +/- 6.7 for all articles, articles accepted for publication and articles rejected, respectively (P = 0.31 between accepted and rejected articles). The mean assessment scores (0-10) were 5.5 +/- 1.5, 5.9 +/- 1.5 and 4.7 +/- 1.4 for all articles, articles accepted for publication and articles rejected, respectively. Accepted articles had a significantly higher assessment score than rejected articles (P = 0.006). For each group, there was a negative correlation between the number of deficiencies and the mean assessment score (P > 0.05). Significantly more rejected articles ' em leader did not further our knowledge' (P = 0.0014) and ' em leader did not describe background information adequately' (P = 0.049). Many rejected articles had ' em leader findings that were not clinically or socially significant' (P = 0.07). Common deficiencies among all articles included ambiguity of the methods (77%) and results (68%), conclusions not warranted by the data (72%), poor referencing (56%), inadequate study design description (51%), unclear tables (49%), an overly long discussion (49%), limitations of the study not described (51%), inadequate definition of terms (49%) and subject selection bias (40%). CONCLUSIONS: Researchers should undertake studies that are likely to further our knowledge and be clinically or socially significant. Deficiencies in manuscript preparation are more frequent than mistakes in study design and execution. Specific training or assistance in manuscript preparation is indicated. PMID- 11903430 TI - Agreement between troponin T levels from plain and heparinized tubes. AB - OBJECTIVE: To determine whether there was a clinically significant effect on troponin T measurement when the sample was collected in a heparinized (plasma) blood collection tube compared with a serum tube. METHODS: Prospective cohort study using a convenience sample of 198 patients with undifferentiated illness presenting to an Emergency Department who required troponin T measurement. Samples were collected in both plain (serum) tubes and plasma tubes for comparison. All samples were measured using an Elecsys 2010 Immunoassay system (Roche-Boehringer Mannheim, Germany). RESULTS: There were 35 troponin T measurements > or = 0.03 microg/L (the limit of reproducibility of the test). The negative predictive value for troponin T performed in heparinized tubes compared with plain tubes was 100% (95% confidence interval 96.4-100) at the > or = 0.03 microg/L level and 100% (95% confidence interval 97-100%) at the > or = 0.1 microg/L level. At a cut-off point for risk stratification in acute coronary syndromes (> or = 0.1 microg/L), there was 100% concordance between the two measurements for each sample. CONCLUSION: The use of plasma (heparinized) tubes for the collection of troponin T samples is unlikely to produce clinically significant false-negative results compared with collection of troponin T samples in serum (plain) tubes. PMID- 11903431 TI - The effect of the InterLink cannula on fluid flow rates and haemolysis. AB - OBJECTIVE: To examine the effect of the InterLink cannula system (Becton Dickinson, New Jersey, NY, USA) on intravenous fluid flow rates and red blood cell haemolysis. METHODS: An in vitro study was performed with crystalloid and packed red blood cells run through intravenous catheters of various sizes, with and without the InterLink cannula. Data recorded and analysed included fluid flow rates and plasma free haemoglobin concentration. RESULTS: The InterLink cannula significantly reduced flow of crystalloid through a 14G catheter (reduction of median flow rate by 0.72 L/h, 12.3%, P < 0.001). There was a reduction of flow of packed red blood cells through a 14G catheter (0.11 L/h) but this was not statistically significant. There was no significant reduction of flow through smaller catheters and no significant effect on red blood cell haemolysis. CONCLUSIONS: The InterLink cannula reduced flow rates through large diameter intravenous catheters. Because of other factors affecting fluid infusion in vivo this is of minor clinical significance. No increased haemolysis of red blood cells occurred with the InterLink cannula. PMID- 11903433 TI - Angioedema in the emergency department: a presentation of lymphoma. AB - A 58-year-old woman presented to emergency departments on several occasions with episodic angioedema. Lymphoplasmacytic lymphoma with an IgM paraprotein (Waldenstrom's macroglobulinemia) was eventually diagnosed 14 months later in association with acquired C1 esterase inhibitor deficiency. Resolution of the angioedema and C1 esterase inhibitor deficiency was achieved with danazol and treatment of the underlying lymphoma. PMID- 11903432 TI - Abstract to publication ratio for papers presented at scientific meetings: how does emergency medicine compare? AB - OBJECTIVES: The aims of the present study were to determine the publication rate of abstracts presented by Australasian emergency physicians at major emergency medicine meetings and to identify the site of publication of papers. METHOD: All free paper abstracts presented (oral and poster) by Australasian emergency physicians and trainees at five Australasian College for Emergency Medicine/Australasian Society for Emergency Medicine and International Conference on Emergency Medicine meetings between 1995 and 1998 were identified retrospectively from conference programmes. In order to determine whether or not the abstract had been published, the PubMed database (http://www4.ncbi.nlm.nih.gov/PubMed/) was searched using the presenter's name and key words from the abstract. In addition, a hand search of the non-abstracted journal Emergency Medicine was conducted. RESULTS: Of the 207 free paper abstracts identified, 73 (35%) had been published as full articles. Papers were published in a variety of journals; however, Emergency Medicine accounted for almost half the published papers. The mean time between presentation and publication was 12.6 months (median 11 months). CONCLUSION: The abstract to publication rate for papers presented by Australasian emergency physicians and trainees at Australasian College for Emergency Medicine/Australasian Society for Emergency Medicine and International Conference on Emergency Medicine meetings is 35%, which is lower than that reported by some other established specialities, but comparable to rates reported for US-based national and international emergency medicine meetings. Future research should look at barriers to the publication of findings and ways to assist the publication process. PMID- 11903434 TI - Recommendations for the management of over-anticoagulation with warfarin. PMID- 11903440 TI - 'Sieve', 'sort' or START. PMID- 11903442 TI - Tracheostomy is the only safe option. PMID- 11903451 TI - Authors' Credentials and Affiliations. PMID- 11903450 TI - CME Overview. PMID- 11903452 TI - Authors' Disclosure Statements. PMID- 11903453 TI - Self-Assessment Quiz. PMID- 11903454 TI - Posttest Answer Sheet/Registration Form. PMID- 11903455 TI - Evaluation Form Matching the Medicine to the Patient. PMID- 11903456 TI - Cardiovascular regulation through hypothalamic GABA(A) receptors in a genetic absence epilepsy model in rat. AB - PURPOSE: Gamma-aminobutyric acid (GABA) plays a vital role in both central cardiovascular homeostasis and pathogenesis of epilepsy. Epilepsy affects autonomic nervous system functions. In this study, we aimed to clarify the role of GABAA receptors in hypothalamic cardiovascular regulation in a genetically determined animal model of absence epilepsy. METHODS: Nonepileptic Wistar rats and genetic absence epilepsy rats from Strasbourg (GAERS) were instrumented with a guide cannula for drug injection and extradural electrodes for EEG recording. After a recovery period, iliac arterial catheters were inserted for direct measurement of mean arterial pressure and heart rate. Bicuculline, a GABA(A) receptor antagonist, was injected into the dorsomedial (DMH) or posterior (PH) hypothalamic nuclei of nonepileptic control rats or GAERS. Blood pressure, heart rate, and EEG recordings were performed in conscious unrestrained animals. RESULTS: Bicuculline injections into the hypothalamus produced increases in blood pressure and heart rate of both control rats and GAERS. The DMH group of GAERS showed a twofold increase in the blood pressure and the heart rate compared with those of control rats. Pressor responses to bicuculline, when microinjected into the PH, were similar in the nonepileptic animals and GAERS. Conversely, the amplitude of tachycardic responses to the administration of bicuculline into the PH was significantly higher in GAERS compared with those of control rats. CONCLUSIONS: The bicuculline-induced increases in blood pressure and heart rate were more prominent when given in the DMH of GAERS. These results indicate an increased GABA(A) receptor-mediated cardiovascular response through the DMH in conscious rats with absence epilepsy. PMID- 11903457 TI - Anticonvulsant profile and teratogenicity of N-methyl-tetramethylcyclopropyl carboxamide: a new antiepileptic drug. AB - PURPOSE: The studies presented here represent our efforts to investigate the anticonvulsant activity of N-methyl-tetramethylcyclopropyl carboxamide (M-TMCD) and its metabolite tetramethylcyclopropyl carboxamide (TMCD) in various animal (rodent) models of human epilepsy, and to evaluate their ability to induce neural tube defects (NTDs) and neurotoxicity. METHODS: The anticonvulsant activity of M TMCD and TMCD was determined after intraperitoneal (i.p.) administration to CF#1 mice, and either oral or i.p. administration to Sprague-Dawley rats. The ability of M-TMCD and TMCD to block electrical-, chemical-, or sensory-induced seizures was examined in eight animal models of epilepsy. The plasma and brain concentrations of M-TMCD and TMCD were determined in the CF#1 mice after i.p. administration. The induction of NTDs by M-TMCD and TMCD was evaluated after a single i.p. administration at day 8.5 of gestation in a highly inbred mouse strain (SWV) that is susceptible to valproic acid-induced neural tube defects. RESULTS: In mice, M-TMCD afforded protection against maximal electroshock (MES) induced, pentylenetetrazol (Metrazol)-induced, and bicuculline-induced seizures, as well as against 6-Hz "psychomotor" seizures and sound-induced seizures with ED50 values of 99, 39, 81, 51, and 10 mg/kg, respectively. In rats, M-TMCD effectively prevented MES- and Metrazol-induced seizures and secondarily generalized seizures in hippocampal kindled rats (ED50 values of 82, 45, and 39 mg/kg, respectively). Unlike M-TMCD, TMCD was active only against Metrazol induced seizures in mice and rats (ED50 values of 57 and 52 mg/kg, respectively). Neither M-TMCD nor TMCD was found to induce NTDs in SWV mice. CONCLUSIONS: The results obtained in this study show that M-TMCD is a broad-spectrum anticonvulsant drug that does not induce NTDs and support additional studies to evaluate its full therapeutic potential. PMID- 11903458 TI - Chromosomal abnormalities and epilepsy: a review for clinicians and gene hunters. AB - PURPOSE: We analyzed databases on chromosomal anomalies and epilepsy to identify chromosomal regions where abnormalities are associated with clinically recognizable epilepsy syndromes. The expectation was that these regions could then be offered as targets in the search for epilepsy genes. METHODS: The cytogenetic program of the Oxford Medical Database, and the PubMed database were used to identify chromosomal aberrations associated with seizures and/or EEG abnormalities. The literature on selected small anomalies thus identified was reviewed from a clinical and electroencephalographic viewpoint, to classify the seizures and syndromes according to the current International League Against Epilepsy (ILAE) classification. RESULTS: There were 400 different chromosomal imbalances described with seizures or EEG abnormalities. Eight chromosomal disorders had a high association with epilepsy. These comprised: the Wolf Hirschhorn (4p-) syndrome, Miller-Dieker syndrome (del 17p13.3), Angelman syndrome (del 15q11-q13), the inversion duplication 15 syndrome, terminal deletions of chromosome 1q and 1p, and ring chromosomes 14 and 20. Many other segments had a weaker association with seizures. The poor quality of description of the epileptology in many reports thwarted an attempt to make precise karyotype phenotype correlations. CONCLUSIONS: We identified certain chromosomal regions where aberrations had an evident association with seizures, and these regions may be useful targets for gene hunters. New correlations with specific epilepsy syndromes were not revealed. Clinicians should continue to search for small chromosomal abnormalities associated with specific epilepsy syndromes that could provide important clues for finding epilepsy genes, and the epileptology should be rigorously characterized. PMID- 11903460 TI - Treatment of refractory status epilepticus with pentobarbital, propofol, or midazolam: a systematic review. AB - BACKGROUND: New continuous infusion antiepileptic drugs (cIV-AEDs) offer alternatives to pentobarbital for the treatment of refractory status epilepticus (RSE). However, no prospective randomized studies have evaluated the treatment of RSE. This systematic review compares the efficacy of midazolam (MDL), propofol (PRO), and pentobarbital (PTB) for terminating seizures and improving outcome in RSE patients. METHODS: We performed a literature search of studies describing the use of MDL, PRO, or PTB for the treatment of RSE published between January 1970 and September 2001, by using MEDLINE, OVID, and manually searched bibliographies. We included peer-reviewed studies of adult patients with SE refractory to at least two standard AEDs. Main outcome measures were the frequency of immediate treatment failure (clinical or electrographic seizures occurring 1 to 6 h after starting cIV-AED therapy) and mortality according to choice of agent and titration goal (cIV-AED titration to "seizure suppression" versus "EEG background suppression"). RESULTS: Twenty-eight studies describing a total of 193 patients fulfilled our selection criteria: MDL (n = 54), PRO (n = 33), and PTB (n = 106). Forty-eight percent of patients died, and mortality was not significantly associated with the choice of agent or titration goal. PTB was usually titrated to EEG background suppression by using intermittent EEG monitoring, whereas MDL and PRO were more often titrated to seizure suppression with continuous EEG monitoring. Compared with treatment with MDL or PRO, PTB treatment was associated with a lower frequency of short-term treatment failure (8 vs. 23%; p < 0.01), breakthrough seizures (12 vs. 42%; p < 0.001), and changes to a different cIV-AED (3 vs. 21%; p < 0.001), and a higher frequency of hypotension (systolic blood pressure <100 mm Hg; 77 vs. 34%; p < 0.001). Compared with seizure suppression (n = 59), titration of treatment to EEG background suppression (n = 87) was associated with a lower frequency of breakthrough seizures (4 vs. 53%; p < 0.001) and a higher frequency of hypotension (76 vs. 29%; p < 0.001). CONCLUSIONS: Despite the inherent limitations of a systematic review, our results suggest that treatment with PTB, or any cIV-AED infusion to attain EEG background suppression, may be more effective than other strategies for treating RSE. However, these interventions also were associated with an increased frequency of hypotension, and no effect on mortality was seen. A prospective randomized trial comparing different agents and titration goals for RSE with obligatory continuous EEG monitoring is needed. PMID- 11903459 TI - Multiple subpial transection for intractable partial epilepsy: an international meta-analysis. AB - PURPOSE: Because the number and variety of patients at any single facility is not sufficient for clinical or statistical analysis, data from six major epilepsy centers that performed multiple subpial transections (MSTs) for medically intractable epilepsy were collected. METHODS: A meta-analysis was performed to elucidate the indications and outcome, and to assess the results of the procedure. Overall, 211 patients were represented with data regarding preoperative evaluation, procedures, seizure types and frequencies before and after surgery, postoperative deficits, and demographic information. Fifty-three patients underwent MST without resection. RESULTS: In patients with MST plus resection, excellent outcome (>95% reduction in seizure frequency) was obtained in 87% of patients for generalized seizures, 68% for complex partial seizures, and 68% for simple partial seizures. For the patients who underwent MST without resection, the rate of excellent outcome was only slightly lower, at 71% for generalized, 62% for complex partial, and 63% for simple partial seizures. EEG localization, age at epilepsy onset, duration of epilepsy, and location of MST were not significant predictors of outcome for any kinds of seizures after MST, with or without resection. New neurologic deficits were found in 47 patients overall, comparable in MST with resection (23%) or without (19%). CONCLUSIONS: These preliminary results suggest that MST has efficacy by itself, with minimal neurologic compromise, in cases in which resective surgery cannot be used to treat uncontrolled epilepsy. MST should be investigated as a stand-alone procedure to allow further development of criteria and predictive factors for outcome. PMID- 11903462 TI - Independent component analysis of ictal EEG in medial temporal lobe epilepsy. AB - PURPOSE: Application of independent component analysis (ICA) to interictal EEGs and to event-related potentials has helped noise reduction and source localization. However, ICA has not been used for the analysis of ictal EEGs in partial seizures. In this study, we applied ICA to the ictal EEGs of patients with medial temporal lobe epilepsy (TLE) and investigated whether ictal components can be separated and whether they indicate correct lateralization. METHODS: Twenty-four EEGs from medial TLE patients were analyzed with the extended ICA algorithm. Among the resultant 20 components in each EEG, we selected components with an ictal nature and reviewed their corresponding topographic maps for the lateralization. We then applied quantitative methods for the verification of increased quality of the reconstructed EEGs. RESULTS: All ictal EEGs were successfully decomposed into one or more ictal components and nonictal components. After EEG reconstruction with exclusion of artifacts, the lateralizing power of the ictal EEG was increased from 75 to 96%. CONCLUSIONS: ICA can separate successfully the manifold components of ictal rhythms and can improve EEG quality. PMID- 11903461 TI - Serum and CSF glutamine levels in valproate-related hyperammonemic encephalopathy. AB - PURPOSE: To investigate ammonia and glutamine levels in valproate (VPA)-related hyperammonemic encephalopathy (VHE). METHODS: We reviewed the medical records and EEG recordings of seven adults diagnosed with VHE. RESULTS: Venous ammonia levels were elevated in five (71%) of the seven patients. Elevated serum or cerebrospinal fluid (CSF) glutamine levels were found in four (80%) of five cases tested, including two who had normal ammonia levels. Initial behavioral signs included violent outbursts in three patients, paranoid ideation severe enough to require restraint in two cases, and milder abnormalities in two instances. The severity of encephalopathy was not related to any particular serum VPA level. In four patients serum VPA levels did not exceed 100 microg/ml, and in one case, VHE developed after taking only one 250-mg dose. Symptoms eventually cleared after reducing the dose of, or discontinuing, VPA. Liver-function tests were normal. Each of six patients tested had EEG findings that supported the diagnosis of VHE and excluded nonconvulsive status epilepticus. The rate of normalization of one patient's serum glutamine level and the EEGs of two cases correlated better with the timing of their delayed clinical recovery than did the more rapid rate of decline of the serum ammonia levels. CONCLUSIONS: Serum or CSF glutamine levels are initially elevated in a majority of patients with suspected VHE, sometimes in the absence of hyperammonemia. Glutamine levels may be useful adjunctive laboratory tests for the diagnosis of VHE. PMID- 11903464 TI - Temporal lobe epilepsy surgery: outcome, complications, and late mortality rate in 215 patients. AB - PURPOSE: We studied the surgical outcome, complications, and the late mortality rate in a large group of patients with medically refractory temporal lobe epilepsy (TLE). METHODS: Two-hundred fifteen patients with TLE were treated surgically between 1984 and 1999 after a comprehensive presurgical evaluation. Patients were followed up at 6 weeks, 3-6 months, and yearly thereafter. In addition, questionnaires were sent on the anniversary of their surgery. Surgical outcome (Engel's classification), complication rate, and factors contributing to late mortality were analyzed. Standardized mortality ratios (SMRs) were calculated. RESULTS: There was no surgical mortality. Two (0.9%) had mild hemiparesis, one (0.4%) had a hemianopia, seven (3.2%) had transient cranial nerve palsies, and eight (3.7%) had transient postoperative language difficulties. One hundred forty-eight (69%) became seizure free, 43 (20%) had rare seizures, 14 (6.5%) had worthwhile seizure reduction, and 10 (4.6%) had no improvement (follow-up, 1-15 years). Three (2%) of 148 seizure-free patients died during follow-up, compared with eight (11.9%) of 67 not seizure-free patients. The mean duration of epilepsy before surgery for the surviving patients was 17.8 years, and for those patients who died, 25.9 years (p < 0.05). Six (5.7%) of 104 patients with right-sided resections died during follow-up, compared with five (4.5%) of 111 with left-sided resections. CONCLUSIONS: Eighty-nine percent of patients became seizure free or had rare seizures, with low morbidity, and no surgical mortality. The late mortality occurred predominantly in patients with persistent seizures (SMR, 7.4). Those patients who died had a longer duration of epilepsy before surgery. In contrast, among those patients who became seizure free, the mortality rate was much lower, and similar to the general population of Indiana (SMR, 1.7). PMID- 11903463 TI - Video-EEG monitoring in the elderly: a review of 94 patients. AB - PURPOSE: We sought to determine the utility and results of video-EEG monitoring in elderly patients. There is an increased incidence of epilepsy in the elderly population. Few studies have assessed the characteristics of epileptic and nonepileptic seizures in this age group. Diagnostic evaluation with video-EEG monitoring is a means to distinguish these different types of events. METHODS: The authors reviewed all patients aged 60 years and older who were admitted to the epilepsy monitoring unit at Columbia-Presbyterian Medical Center from January 21, 1991, to April 12, 1999. RESULTS: A total of 94 patients and 99 patient admissions were identified, accounting for 8% of all admissions. The average age was 70 years, and the mean length of stay was 3.8 days. Typical events were recorded in 75 of the 99 patient admissions. A total of 118 seizures was recorded in 46 patients, and 98 nonepileptic events were seen in 27 patients. Of the patients with nonepileptic events, 13 had psychogenic seizures. The majority of patients with nonepileptic events were taking antiepileptic medication. Whereas 76% of the patients with epileptic events had interictal epileptiform discharges, 26% of the patients with nonepileptic events had epileptiform discharges as well. CONCLUSIONS: Video-EEG monitoring in the elderly leads to a definitive diagnosis in the majority of patients in a relatively short time. Interictal recordings are inadequate in determining the nature of paroxysmal events. Nonepileptic events are common in the elderly, including psychogenic seizures, and these are often misdiagnosed and mistreated as epileptic seizures. PMID- 11903465 TI - Add-on phenytoin fails to prevent early seizures after surgery for supratentorial brain tumors: a randomized controlled study. AB - PURPOSE: To determine the potential effectiveness of phenytoin (PHT) in preventing early postoperative seizures in patients undergoing craniotomy for supratentorial brain tumors. METHODS: Two hundred patients requiring elective craniotomy for supratentorial brain tumors were randomized to two groups of equal size, with a prospective, open-label, controlled design. One group received PHT (18 mg/kg as an intravenous intraoperative load, followed by additional daily doses aimed at maintaining serum PHT concentrations within the 10- to 20-aeg/ml range) for 7 consecutive days. In the other group, PHT was not administered. More than 90% of patients in both groups continued to take preexisting anticonvulsant medication (AEDs) with carbamazepine or phenobarbital throughout the study. The primary efficacy end point was the number of patients remaining free from seizures during the 7-day period after the operation. RESULTS: Of 100 patients allocated to PHT, 13 experienced seizures during the 7-day observation period, compared with 11 of 100 patients in the placebo group (p > 0.05). Most seizures occurred in the first day after surgery in both groups. There were no differences between groups in the proportion of patients experiencing more than one seizure, but there was a trend for generalized seizures to be more common in PHT-treated patients than in controls (11 vs. five patients, respectively). Status epilepticus occurred in one patient in the PHT group and in two patients in the control group. Of the 13 PHT-treated seizure patients, 11 had serum PHT concentrations within the target range, and only two had concentrations below range on the days their seizures occurred. CONCLUSIONS: PHT, given at dosages producing serum concentrations within the target range, failed to prevent early postoperative seizures in patients treated with concomitant AEDs. Prophylactic administration of PHT cannot be recommended in these patients. PMID- 11903466 TI - Epidemiologic features of infantile spasms in Slovenia. AB - PURPOSE: To determine the epidemiologic features of infantile spasms (ISs) in Slovenia. METHODS: Medical records of all children with ISs in Slovenia in the period from 1985 to 1995, based on community pediatrician referrals to four hospitals, including all pediatric EEG laboratories, were retrospectively studied. The outcome was assessed by a follow-up study in 1998. RESULTS: Forty seven children with ISs were identified over an 11-year interval. The cumulative incidence was 2.06 per 10,000 live births. Among 29 (61.7%) children with symptomatic etiology, 14 cases had prenatal etiology [tuberous sclerosis (TS), seven; vascular insult, three; cerebral malformations, three; Down syndrome, one child], 14 perinatal and one postnatal cause: anoxic brain damage after cardiac surgery. Cryptogenic and idiopathic etiology were diagnosed in 13 (27.6%) and five (10.6%) of 47 cases, respectively. The age of onset of ISs ranged from 2 to 10 months. As initial treatment, steroids were used in 19 children (remission in 10); vigabatrin in seven (remission in four), and other antiepileptic drugs (AEDs) in 20 children (remission in six). According to the follow-up study, 18 (38.3%) children were seizure free, and 14 of them had normal mental development. Among 29 mentally retarded children (14 severely), 26 belonged to the symptomatic group. Four children died. CONCLUSIONS: The incidence of ISs in Slovenia is similar to that in some parts of the United States, but lower than that in Finland or Sweden. The outcome depends mainly on etiology. Additional neuroimaging studies are needed for evaluation of cryptogenic cases. PMID- 11903467 TI - Dissociative and associated psychopathological symptoms in patients with epilepsy, pseudoseizures, and both seizure forms. AB - PURPOSE: A controversy currently exists regarding the significance of dissociation and conversion in the pathogenesis of pseudoepileptic seizures. After the abolition of the term "hysterical neurosis" from the current diagnostic systems, these seizures were diagnosed as either Dissociative Disorders (ICD-10) or in the DSM IV as Somatoform disorder, most often of conversion type. Recent studies of patients with Dissociative Disorders found that most patients also had conversion symptoms. METHODS: In the present study, 60 patients of an outpatient clinic for epilepsy were assessed for the presence of dissociative symptoms and general psychopathologic symptoms by using the German version of the Dissociative Experience Scale (DES) and the Symptom Check List (SCL-90-R). RESULTS: The patients with pseudoepileptic seizures showed a significantly higher incidence of dissociation (p < 0.0098) and general psychopathologic symptoms (p < 0.0083). Depression, anxiety, and obsession were dominating psychopathologic symptoms in all patients. CONCLUSIONS: The significantly higher incidence of dissociation in the patients with pseudoepileptic seizures suggests dissociation in the pathogenesis of these seizures. PMID- 11903468 TI - Psychiatric comorbidity and hostility in patients with psychogenic nonepileptic seizures compared with somatoform disorders and healthy controls. AB - PURPOSE: To investigate the prevalence of psychiatric comorbidity and level of anxiety, depression, and aggression in patients with psychogenic nonepileptic seizures compared with those in patients with somatoform disorders and healthy controls. METHODS: Twenty-three patients with psychogenic nonepileptic seizures (PNESs) and 23 age- and sex-matched patients with somatoform disorders (SDs) underwent a clinical and a semistructured psychiatric interview (MINI) and filled in the Hospital Anxiety and Depression scale (HAD) and the Aggression Questionnaire (AQ). Twenty-three sex- and age-matched controls without psychopathology also underwent a clinical interview and completed the HAD and AQ. RESULTS: PNES reported more minor head injuries in the past than did the two comparison groups, and more unspecific EEG dysrhythmias were observed on EEG. Twenty-one PNES patients and 18 with SDs had comorbid psychiatric diagnoses. However, the mean number of comorbid psychiatric diagnoses was higher in the PNES group (1.9 +/- 0.3 compared with 1.5 +/- 0.5 in the SD group; p = 0.003). Ten PNES patients additionally had a somatoform pain disorder, and seven had an undifferentiated somatoform disorder. Both patient groups reported significantly higher levels of anxiety, depression, and anger than did the healthy controls, but the PNES patients had significantly higher level of hostility than both comparison groups. CONCLUSIONS: Increased psychiatric comorbidity is known to be associated with poorer response to regular interventions, and hostility is associated with more hostile coping patterns, often interfering with treatment compliance. Thus the increased prevalence of soft neurologic signs and comorbid psychiatric disorders and increased hostility as well in the PNES group, emphasizes that assessment and treatment of patients with PNES referred to a tertiary center requires an integrated approach involving both neurologic and psychiatric resources. PMID- 11903469 TI - Agranulocytosis associated with lamotrigine in a patient with low-grade glioma. AB - PURPOSE: To report agranulocytosis associated with lamotrigine (LTG) in a patient with a brain tumor. METHODS: A 59-year-old woman with a low-grade glioma and difficult-to-control partial seizures developed agranulocytosis between 9 and 14 weeks after starting LTG. The patient underwent chemotherapy 2 years previously. RESULTS: After stopping LTG, the agranulocytosis persisted for 9 days despite 7 days of granulocyte-colony-stimulating factor (G-CSF), and ultimately resolved. CONCLUSIONS: In this case, the use of LTG was associated with the development of agranulocytosis, and it is likely that this association was causative. PMID- 11903470 TI - Lidocaine-dependent early infantile status epilepticus with highly suppressed EEG. AB - We report an early infantile patient characterized by intractable hyponatremia, progressive megalencephaly, and epileptic seizures with an EEG pattern that alternated between interictal low-voltage background and ictal burst activity. Repeatedly all the abnormal findings improved in a lidocaine-dependent manner. Given the pharmacologic mechanisms of lidocaine as a sodium channel blocker, we speculate that our patient had a sodium channel dysfunction. PMID- 11903471 TI - Fenfluramine as a potential antiepileptic drug. PMID- 11903472 TI - Alfentanil-induced epileptiform activity. PMID- 11903475 TI - Basic mechanisms of psychotropic drugs. AB - Many epilepsy patients, particularly those with complex partial seizures, also develop psychiatric disorders during the course of their illness and have to be treated with psychotropic drugs in addition to their antiepileptic medication. However, the brains of epileptic patients can be considered pathologically altered and psychotropic drugs may thus have profound and stronger effects on seizure threshold or unwanted side effects than in purely psychiatric patients. Thus, the knowledge of the mechanisms of psychotropic drugs is necessary to predict their effects in epilepsy patients. In this review, current concepts of the mechanisms of neuroleptic, antidepressant, and anxiolytic drugs emerging from basic and preclinical research are summarized, and the potential impact of using these drugs in epilepsy patients is discussed. PMID- 11903476 TI - Atypical antipsychotics and serotoninergic antidepressants in patients with epilepsy: pharmacodynamic considerations. AB - PURPOSE: To discuss the pharmacodynamic aspects of the administration of atypical antipsychotics (APs) and serotoninergic antidepressants (SSRIs) to patients with epilepsy. METHODS: This article represents an overview of all studies concerning the administration of APs and SSRIs to people with epilepsy. In particular, it deals with the relationship between neuroleptics (NLTs), APs, SSRIs, serotonin, and dopamine, with special focus on the possible epileptogenic role of psychoactive drugs. RESULTS: NLTs may induce seizures by blocking D2, H1, and.1 receptors, or by sexual hormone activation or a pharmacologic kindling mechanism. The difference among APs in their ability to induce seizures is related mainly to the percentage of D2-receptor occupancy and possibly also to their action on neurosteroids. Seizures occur at SSRIs therapeutic doses, with a 0.1-4% incidence. Coversely, in animal studies fluoxetine was claimed to exert an anticonvulsant action. CONCLUSIONS: The study of the pharmacodynamic aspects of the administration of APs and SSRIs to patients with epilepsy can help to evaluate the importance of some mechanisms of action of several psychoactive drugs in relation to their pro- or anticonvulsant activity. PMID- 11903477 TI - Antidepressant drugs: indications and guidelines for use in epilepsy. PMID- 11903478 TI - Antipsychotic drugs and epilepsy: indications and treatment guidelines. PMID- 11903479 TI - On the use of tranquillisers in epilepsy. AB - In this article, the use of benzodiazepines (BZDs) in epilepsy is reviewed. In particular, it concentrates on the use of oral BZDs, and, following a brief discussion as to their value in psychiatric disorders generally, their more specific use in epilepsy is then noted. The main oral drugs are the 1,4BZDs and 1,5-BZDs. The latter is represented only by clobazam (CLB), and the advantages of the 1,5-BZDs over the 1,4-BZDs are reviewed. It is concluded that CLB has a significant value in a number of situations. It is a first-line add-on therapy when standard medications have failed. It is useful in patients that have catamenial seizures, clusters of seizures, and also in the prevention of postictal syndromes. It is used in childhood epilepsy, and is also helpful in the management of febrile seizures. PMID- 11903480 TI - Psychostimulants and epilepsy. AB - PURPOSE: The aim of this article is to review the literature on the effects of psychostimulants in epileptic subjects in order to reach a consensus statement regarding the use or abuse of these substances. METHODS: Psychostimulant substances have been considered the drugs that share the ability to produce excitation of the CNS leading to convulsions. The stimulation may be at cortical, brainstem, or spinal levels. In this article, the following cortical stimulants are analyzed and discussed: cocaine, amphetamine and related agents, caffeine, cannabinoids, and psychedelic drugs. This review is based on research done using pharmacological textbooks and Medline. RESULTS: The use of cocaine is associated with the occurrence of seizures. The reported frequency varies from 1% to 40% of addicted subjects, based on the typology of the considered study. Amphetamines and related drugs rarely induce epileptic seizures at therapeutic doses, but seizures may occur after the first dosing. Caffeine at high doses may induce epileptic seizures because of its adenosine receptor-antagonizing properties. Marijuana, at variance with other psychostimulants, owing to its serotonin mediated anticonvulsant action, could have a medical use for the treatment of epilepsy. Psychedelic compounds rarely induce epileptic seizures, but the most common clinical CNS complication after ingestion of ecstasy is the occurrence of seizures. CONCLUSIONS: The use of psychostimulants, except for marijuana, can induce single or multiple seizures in healthy subjects. PMID- 11903481 TI - The use of antidepressant and antipsychotic drugs in elderly epilepsy patients. PMID- 11903482 TI - Clinical significance of pharmacokinetic interactions between antiepileptic and psychotropic drugs. AB - As antiepileptic drugs (AEDs) and psychotropic agents are increasingly used in combination, the possibility of pharmacokinetic interactions between these compounds is relatively common. Most pharmacokinetic interactions between AEDs and psychoactive drugs occur at a metabolic level, and usually involve changes in the activity of the cytochrome P450 mixed-function oxidases (CYP) involved in their biotransformation. As a consequence of CYP inhibition or induction, plasma concentrations of a given drug may reach toxic or subtherapeutic levels, and dosage adjustments may be required to avoid adverse effects or clinical failure. Enzyme-inducing AEDs, such as carbamazepine (CBZ), phenytoin (PHT), and barbiturates, stimulate the oxidative biotransformation of many concurrently prescribed psychotropics. In particular, these AEDs may decrease the plasma concentrations of tricyclic antidepressants, many antipsychotics, including traditional compounds, i.e., haloperidol and chlorpromazine, and newer agents, i.e., clozapine, risperidone, olanzapine, quetiapine, and ziprasidone, and some benzodiazepines. Conversely, new AEDs appear to have a lower potential for interactions with all psychotropic drugs. While antipsychotics and anxiolytics do not significantly influence the pharmacokinetics of most AEDs, some newer antidepressants, such as viloxazine, fluoxetine, and fluvoxamine, may lead to higher serum levels of some AEDs, namely CBZ and PHT, through inhibition of CYP enzymes. No significant pharmacokinetic interactions have been documented between AEDs and lithium. Information about CYP enzymes responsible for the biotransformation of individual agents and about the effects of these compounds on the activity of specific CYP enzymes may help in predicting and avoiding clinically significant interactions. Apart from careful clinical observation, serum level monitoring of AEDs and psychotropic drugs can be useful in determining the need for dosage adjustments, especially if there is any change in seizure control, or possible toxicity. PMID- 11903483 TI - When is it inappropriate to prescribe psychotropic medication? AB - Psychotropic medication can be of great value in the treatment of people with epilepsy, but there are situations in which it is not appropriate and other strategies are needed. A number of different strategies, usually not involving psychotropic medication, are required for the behavioural disturbance associated with prodromal mood changes, anxiety-provoking auras, focal discharges, frequent absence seizures, adverse reactions to antiepileptic medication or adverse reactions to antiepileptic drug interactions, difficulties arising from uneven cognitive profiles, reactions to the epilepsy, and causes that are not related to the epilepsy. The first step should always be to determine the cause or causes of behavioural disturbance. A systematic approach to the assessment of the behavioural or psychiatric problems, using a structured diagnostic framework of possible causes, provides the basis for correct diagnosis and management. This systematic approach assists the clinician in deciding when other strategies are preferable to the prescription of psychotropic medication. PMID- 11903484 TI - Post-prandial hypertriglyceridemia and endothelium-dependent vasorelaxation. PMID- 11903485 TI - Advanced glycation and the immune system: stimulation, inhibition or both? PMID- 11903486 TI - Endogenous anti-inflammatory response after coronary injury in a porcine model. AB - BACKGROUND: Corticotropin-releasing hormone (CRH)/adrenocorticotropic hormone (ACTH)/cortisol is the major anti-inflammatory system. After percutaneous translumenal angioplasty, an inflammatory process is triggered. We investigate whether CRH/ACTH/cortisol axis is activated after deep vessel wall injury (DVWI). MATERIALS AND METHODS: Plasma and leukocyte CRH and ACTH, serum cortisol and IL 1beta, and leukocyte cAMP were measured (ELISA) in 16 pigs after anaesthesia (baseline), 60 min into anaesthesia without causing vascular injury and 90 min after DVWI of the left anterior descending (LAD) coronary artery induced by percutaneous directional atherectomy (Atherocath GTO 7F; DVI, Inc., Temecula, USA). Biochemical variables were also measured at baseline, 60 and 180 min into anaesthesia in six additional pigs without coronary intervention. RESULTS: MANOVA showed that CRH/ACTH/Cortisol, cAMP and IL-1beta production was not modified during anaesthesia. Post-DVWI plasma CRH (0.077 +/- 0.046 ng mL-1), and cellular cAMP (0.14 +/- 0.067 pmol 10(-6) cells) increased significantly (P = 0.001) with respect to their baseline values (CRH = 0.036 +/- 0.013 ng mL-1; cAMP = 0.081 +/- 0.034 pmol 10-6). There was also a statistically significant increase (P = 0.02) in post-DVWI IL-1beta (from 46.6 +/- 12.8 to 64.05 +/- 13.5 pg mL-1), and in serum cortisol (P = 0.05) compared to its baseline values (8.98 +/- 3.2 microgr dL-1 vs. 6.57 +/- 2.3 microgr dL-1, respectively). CONCLUSION: In our experimental model, coronary vessel wall injury-activated CRH/ACTH/cortisol axis caused a significant increase in plasma CRH, cortisol and cellular cAMP levels, which may influence the response of coronary arteries to injury. PMID- 11903487 TI - Effects of short-term feeding of a highly palatable diet on vascular reactivity in rats. AB - BACKGROUND: Numerous studies have shown that long-term consumption of high-fat, high-energy diet results in obesity, which in turn, leads to cardiovascular disorders. However, there is little or no data on the acute effects of a highly palatable diet on vascular function. MATERIAL AND METHODS: In this study we aimed to evaluate the changes in metabolic and vascular reactivity in Wistar rats fed a palatable diet for 2 days. RESULTS: Two-days feeding of rats with a palatable diet did not effect body weight, fat-pad mass or gastrocnemius muscles weights. Nor there were any changes in plasma glucose, insulin or leptin levels. However, compared with chow-fed rats, palatable diet-fed rats had significantly raised plasma free fatty acids and triglycerides levels (for both, P < 0.01). Compared with chow-fed animals, vasorelaxation responses to carbamylcholine and sodium nitroprusside were significantly attenuated in palatable diet-fed rats (for both, P < 0.01). However, there were no differences in histamine-induced vasorelaxation between chow-fed and palatable diet-fed rats. CONCLUSION: These data indicates that diet-induced endothelium-dependent and -independent vascular dysfunction occurs long before obesity develops. PMID- 11903488 TI - Renal proximal tubular cell growth and differentiation are differentially modulated by renotropic growth factors and tyrosine kinase inhibitors. AB - BACKGROUND: The renotropic growth factors (GFs), hepatocyte GF (HGF), epidermal GF (EGF), and insulin-like GF-I (IGF-I) accelerate renal regeneration in animal models after toxic or ischemic injury. These GFs initiate their biological effects on renal tubular cells by interaction with specific transmembrane receptor tyrosine kinases. MATERIALS AND METHODS: In the proximal tubular cell line PT-1, the biological effects of HGF, EGF, and IGF-I and the growth inhibitory effects of different tyrosine kinase inhibitors (TKIs) were investigated. Receptor binding and tyrosine kinase phosphorylation were determined by ligand binding studies and Western blot analysis. RESULTS: HGF, EGF, and IGF-I bound with nanomolar affinity to their specific cell membrane receptor tyrosine kinases. In contrast to EGF or IGF-I, HGF induced a variety of cell morphological changes, including cell scattering, formation of tubular structures, and expression of long microvilli on the apical cell membrane. HGF was a 10-fold more potent and more effective growth promoter than EGF or IGF-I. Among the TKIs tested, the mitogenic effect of HGF could be more specifically inhibited by emodin and tyrphostin, that of EGF by methyl-2,5-dihydroxycinnamate, lavendustin A, and genistein, and that of IGF-I by geldanamycin. CONCLUSIONS: In contrast to EGF and IGF-I, HGF stimulated both growth and differentiation of renal proximal tubular cells, demonstrating the amazing biological potency of this renotropic growth factor. Selective TKIs may be a promising approach to modulate diseases with abnormalities in protein kinase signalling pathways such as renal cell carcinoma. PMID- 11903490 TI - The improvement of renal concentration capacity after surgery for primary hyperparathyroidism. AB - BACKGROUND: Improvement of renal concentration capacity was long ago shown to occur after surgery for primary hyperparathyroidism (pHPT). Study of concentration capacity is of interest, as it was also shown to be a predictive factor for the risk of death in patients with pHPT, and it affected the risk of death independently of 33 other variables in multivariate analysis. METHODS: There were 98 patients with verified pHPT operated on in the years 1958-81, who had urine osmolality determinations performed both before and after surgery: 63 immediately after, and 35 with mean 3.9 years delay (SD = 1.8). Another seven patients with pHPT had urine osmolality determinations performed preoperatively only. Non-parametric sign tests, regression analysis, and correlation tests were performed. RESULTS: Both patients with severe or moderate, and mild pHPT showed a substantial change of renal concentration capacity, with mean increase of 28.3% (SD = 28.4). The increase generally occurred soon after surgery. In eight out of 98 patients, there was no improvement. A relationship was found between improvement and preoperative peak serum calcium level. In seven out of seven patients followed, untreated for mean 5.3 years (SD = 3.2), there was a mean 15% (SD = 8.0) deterioration of renal concentration capacity. CONCLUSIONS: The findings of this study add cause for surgery in patients with pHPT and give no reason for different treatment of severe, moderate or mild disease. PMID- 11903489 TI - Food deprivation increases post-heparin lipoprotein lipase activity in humans. AB - OBJECTIVE: To study the effect of fasting on lipoprotein lipase (LPL) activity in human post-heparin plasma, representing the functional pool of LPL. DESIGN: Fourteen healthy volunteers were recruited for the study. The subjects were fasted for 30 h. Activities of LPL and hepatic lipase (HL), and LPL mass, were measured in pre- and post-heparin plasma in the fed and in the fasted states, respectively. For comparison, LPL and HL activities were measured in pre- and post-heparin plasma from fed and 24-h-fasted guinea pigs. RESULTS: Fasting caused a significant drop in the levels of serum insulin, triglycerides and glucose in the human subjects. Post-heparin LPL activity increased from 79 +/- 6.4 mU mL-1 in the fed state to 112 +/- 10 mU mL-1 in the fasted state (P < 0.01), while LPL mass was 361 +/- 29 in the fed state and 383 +/- 28 in the fasted state, respectively (P = 0.6). In contrast, fasting of guinea pigs caused an 80% drop in post-heparin LPL activity. The effect of fasting on human and guinea pig post heparin HL activity were moderate and statistically not significant. CONCLUSIONS: In animal models such as rats and guinea pigs, post-heparin LPL activity decreases on fasting, presumably due to down-regulation of adipose tissue LPL. In humans, fasting caused increased post-heparin LPL activity. PMID- 11903491 TI - Re-adaptation of the gastroduodenal mucosa to DNA synthesis during protracted stress. AB - BACKGROUND: Previous study showed that under the influence of protracted physical stress, the DNA synthesis (DS) of the gastroduodenal mucosa rises, the highest peak being reached at 4 weeks. At 8 stress weeks, the DS decreases to values similar to those recorded at the beginning of the experiment. The possibility that this DS adaptation (DSA) could be maintained beyond 8 weeks in animals allowed to subsequent resting (stress-free) weeks, was now explored. METHODS: Sixty-five rats were investigated. Sixty rats were transported to the stress laboratory. Thirty were water plunged and 30 sham handled once a day, 5 days a week, for 8 weeks. After 8 stress weeks, groups of 5 animals were allowed to rest for one, two, three and four weeks, ending with single water plunging or sham handling. All 65 animals received ip injection of 3H-thymidine before they were sacrificed. The ratio radioactive DNA/total DNA reflected the DS of the stomach and duodenum. RESULTS: DSA was achieved at 8 stress weeks (stomach). The DS in the stomach of stressed rats had significantly increased at 10 weeks (P < 0.05), but at 12 weeks it had decreased to 8 weeks values. The DS in the duodenum of stressed rats had significantly increased at 9, 10 and 11 weeks (P < 0.05), but at 12 weeks DS values had decreased to those of 8 weeks. CONCLUSIONS: The DSA at 8 stress weeks had not prevailed because the DS increased considerably in the following weeks. At 9-11 weeks, DS values had significantly increased (P < 0.05) in stressed rats, but at 12 weeks a DS re-adaptation (DSRA) had occurred. Thus, it took 8 weeks to achieve DSA (stomach), but only 4 weeks to accomplish DSRA. Animals experienced sham handling as an stressor (milder) as it also induced DS re-adjustments in the gastroduodenal mucosa. Autoradiography showed that the labelling in DNA synthesizing cells was limited almost exclusively to the mucosal layer. The model described may prove of value to studies aimed at abating the disparate fluctuations of DNA synthesis in the gastroduodenal mucosa during the various phases of protracted stress. PMID- 11903493 TI - In vitro effect of advanced glycation end-products on human polymorphonuclear superoxide production. AB - BACKGROUND: Advanced glycation end-products (AGEs) are elevated in the sera of diabetic patients. The latter are prone to severe bacterial infections. Advanced glycation end-products have been shown to modulate immune competent cell activities. In this study we examined the in vitro effect of advanced glycation end-products on superoxide anion generation by human polymorphonuclear leukocytes. MATERIALS AND METHODS: Advanced glycation end-products were prepared by incubation of bovine serum albumin (BSA) with glucose for 90 days. Superoxide production was measured as the superoxide dismutase-inhibitable reduction of ferricytochrome c. The effect of advanced glycation end-products on superoxide production was evaluated in both baseline (nonstimulated) and stimulated (by either formyl-methionyl-leucyl-phenylalanine, or phorbol-myristate-acetate) polymorphonuclear leukocytes. RESULTS: The baseline superoxide production of polymorphonuclear leukocytes was significantly increased by advanced glycation end-products in a dose-dependent manner. In contrast, in stimulated polymorphonuclear leukocytes advanced glycation end-products significantly inhibited superoxide production, again in a dose-dependent manner. This inhibitory effect of advanced glycation end-products was observed after dialyzing AGE-BSA, thereby eliminating the possible influence of reactive carbohydrates. No modification of superoxide production was seen with BSA and only a mild inhibitory effect of glucose at high concentrations. CONCLUSIONS: Advanced glycation end-products depress superoxide production by stimulated polymorphonuclear leukocytes. As superoxide plays an essential role in bactericidal activity, this polymorphonuclear leukocyte dysfunction may be a contributory factor to the increased prevalence and severity of bacterial infection seen in diabetic patients. PMID- 11903492 TI - Butyrate and the cytokine-induced alpha1-proteinase inhibitor release in intestinal epithelial cells. AB - BACKGROUND: Alpha1-proteinase inhibitor (alpha1-PI), an anti-inflammatory protein thought to play a role in the intestinal inflammation, is synthesised by and released from the intestinal epithelial cells. IL-1beta is a key proinflammatory cytokine in the abnormal immune response that occurs in inflammatory bowel disease. Butyrate is a normal luminal constituent in the colon, known to be of benefit in preventing inflammatory bowel disease. Direct modes of action of butyrate in intestinal inflammation have been poorly studied so far. The aim of this study was to investigate the effects of butyrate on cytokine-mediated alpha1 PI release in intestinal epithelial cells. METHODS: Differentiated Caco-2 cells were incubated with IL-1beta in the presence or absence of 2 mM butyrate. Alpha1 PI expression in the cells was evaluated by Western blot analysis and alpha1-PI release by ELISA. RESULTS: Treatment with butyrate alone had no effect on alpha1 PI expression in differentiated Caco-2 cells. However, treatment of the cells with 2 mM butyrate significantly reduced the alpha1-PI level in IL-1beta-treated cells. In the cell culture medium, the presence of butyrate impaired the IL-1beta induced alpha1-PI release to 17-35%. The treatment induced no change in the number of detached cells or the percentage of viable cells. CONCLUSION: Our data show that butyrate inhibits alpha1-PI release from Caco-2 colonocytes treated with IL-1beta. It is therefore likely that anti-inflammatory actions of butyrate occur via a mechanism that does not involve direct regulation of cytokine-induced anti-inflammatory protein expression in intestinal epithelial cells. PMID- 11903494 TI - Serum amyloid A and high-density lipoprotein cholesterol: serum markers of inflammation in sarcoidosis and other systemic disorders. AB - Hypocholesterolemia has been observed in several inflammatory diseases such as rheumatoid arthritis, myeloproliferative disorders, systemic lupus erythematosus and sarcoidosis. Serum amyloid A is an acute-phase reactant that is related to the high-density lipoprotein cholesterol. This review discusses the relationship between the activation of the cells of the monocyte-macrophage system, determined by the serum amyloid A levels, and the lipid metabolism, measured as alterations in plasma lipoprotein concentrations. The mechanisms of this association during acute inflammation are also discussed in this review. PMID- 11903495 TI - Mixed cryoglobulinemia is associated with increased risk for death, or neoplasia in HIV-1 infection. AB - BACKGROUND: Cryoglobulinemia has been reported in several chronic infectious and autoimmune diseases, and in patients with HIV-1 infection. Cryoglobulinemia associated with hepatitis C virus infection is considered a risk factor for the development of neoplasia, especially B-cell non-Hodgkin lymphoma. This study was undertaken to investigate whether the presence of circulating cryoglobulins is associated with survival or development of neoplastic disease in HIV-1 infection. DESIGN: We evaluated 87 unselected consecutive HIV-1 infected patients for the presence of cryoglobulinemia and they were prospectively followed up for a median of 34 months, with clinic visits at 4-month intervals. None of the patients had neoplasia at study entry. Time-to-event analysis for death, neoplasm and B-cell lymphoproliferative disorder were performed with Cox proportional hazards models. RESULTS: Mixed cryoglobulinemia (types II and III) was detected in 24 (28%) of the 87 patients. During the follow up, 12 patients died and 8 developed neoplastic disease. Multivariate analysis showed that circulating cryoglobulins were an independent predictor of death [relative risk (RR), 4.97; 95% confidence intervals (CI), 1.26-19.63] and development of neoplasia (RR, 5.18; 95% CI, 1.23 21.83). In addition, cryoglobulinemia reached borderline significance as a predictor of lymphoproliferative disorder of B-cell origin (P = 0.08; RR, 4.53; 95% CI, 0.83-24.75). CONCLUSIONS: Our results suggest that cryoglobulinemia is associated with an increased risk for death, neoplasia or development of lymphoproliferative disorder of B-cell origin, in HIV-1 infected patients. PMID- 11903498 TI - Complement and its implications in cardiac ischemia/reperfusion: strategies to inhibit complement. AB - Although reperfusion of the ischemic myocardium is an absolute necessity to salvage tissue from eventual death, it is also associated with pathologic changes that represent either an acceleration of processes initiated during ischemia or new pathophysiological changes that were initiated after reperfusion. This so called "reperfusion injury" is accompanied by a marked inflammatory reaction, which contributes to tissue injury. In addition to the well known role of oxygen free radicals and white blood cells, activation of the complement system probably represents one of the major contributors of the inflammatory reaction upon reperfusion. The complement may be activated through three different pathways: the classical, the alternative, and the lectin pathway. During reperfusion, complement may be activated by exposure to intracellular components such as mitochondrial membranes or intermediate filaments. Two elements of the activated complement contribute directly or indirectly to damages: anaphylatoxins (C3a and C5a) and the membrane attack complex (MAC). C5a, the most potent chemotactic anaphylatoxin, may attract neutrophils to the site of inflammation, leading to superoxide production, while MAC is deposited over endothelial cells and smooth vessel cells, leading to cell injury. Experimental evidence suggests that tissue salvage may be achieved by inhibition of the complement pathway. As the complement is composed of a cascade of proteins, it provides numerous sites for pharmacological interventions during acute myocardial infarction. Although various strategies aimed at modulating the complement system have been tested, the ideal approach probably consists of maintaining the activity of C3 (a central protein of the complement cascade) and inhibiting the later events implicated in ischemia/reperfusion and also in targeting inhibition in a tissue-specific manner. PMID- 11903496 TI - Incidence and geographical distribution of sudden infant death syndrome in relation to content of nitrate in drinking water and groundwater levels. AB - BACKGROUND: Previous studies indicate that the enteral bacterial urease is inhibited in victims of sudden infant death syndrome (SIDS). One possible inhibitor of this bacterial activity is nitrate. If ambient pollution by nitrate is involved in the etiology of SIDS only a fraction of the nitrate concentration not infrequently found in drinking water would be enough for this inhibition. METHODS: Occurrence of SIDS (n = 636) in Sweden during the period 1990 through 1996 were analysed regarding geographical and seasonal distribution in relation to the nitrate concentration in drinking water and changes in the groundwater level. RESULTS: Both the birth rate and the incidence of SIDS decreased during the study period. One quarter of the municipalities constituting 11% of the population had no cases, the maximum incidence being 6.5 per 1000 live births. Seasonality: The northernmost parts of the country had its highest incidence when the rest of the country had its lowest incidence, and the occurrence of individual deaths was associated with the recharge of groundwater which increases its nitrate content. The local incidence of SIDS was correlated (rs = 0.34-0.87) to maximally recorded concentrations of nitrate in drinking water. CONCLUSIONS: The seasonal distribution of SIDS was widely different from the south to the north of the country and seems to be associated with differences in the groundwater level changes subsequent to precipitation, frost penetration, and melting of snow. Use of drinking water with high peak concentrations or great variations in nitrate concentration was correlated to the incidence of SIDS. PMID- 11903499 TI - Inhibition of rat parotid and submandibular gland functions by ofloxacin, a fluoroquinolone antibiotic. AB - While fluoroquinolones are widely used in the treatment of various infectious disease, not enough attention has been paid to their adverse effects on salivary glands functions. In the present study, the effects of ofloxacin, a fluoroquinolone antibiotic, on rat parotid and submandibular gland functions, were examined in an acute experiment. Ofloxacin (OFLX) was administered intraperitoneally at various doses (20, 40, and 80 mg/kg). Pure parotid and submandibular saliva were collected intraorally by microployethylene tubes under anaesthesia using a dissecting microscope. Flow rate, amylase activity, total protein, and calcium concentrations were reduced by all doses of OFLX (20, 40, 80 mg/kg, P < 0.01) in parotid saliva. In parotid saliva, sodium and potassium were increased by doses of 20 and 40 mg/kg (P < 0.01) and 20, 40, 80 mg/kg (P < 0.01) respectively. In submandibular saliva, flow rate, total protein and calcium concentrations were significantly reduced by all doses (20, 40, 80 mg/kg, P < 0.01). Sodium and potassium concentrations were also increased by a dose of 80 mg/kg, (P < 0.05) in submandibular saliva. It is concluded that ofloxacin inhibits rat salivary gland functions, which might be observed as a side-effect in humans. Properties of fluoroquinolones to alter intracellular cAMP and calcium levels and their ability to suppress DNA, RNA and protein synthesis of acinar cells might be possible reasons for the observed changes. PMID- 11903501 TI - Role of endothelium in adenosine receptor-mediated vasorelaxation in hypertensive rats. AB - The present study was designed to investigate the role of endothelium derived relaxing factor nitric oxide (NO) in adenosine A2 receptor mediated vasorelaxation in normotensive (WKY) and hypertensive (SHR) rat aortic ring preparations. Adenosine analogues, 2-chloroadenosine (CAD) and 5 ethylcarboxamidoadenosine (NECA) produced concentration-dependent (10(-9)-10(-4) M) relaxation in phenylephrine (1 x 10(-6) M) precontracted vascular rings, which was significantly shifted to the right in SHR compared to WKY rats. Endothelium removal attenuated CAD and NECA relaxation responses in both SHR and WKY and abolished the difference in relaxation between SHR and WKY vascular tissues. The relaxation response to CAD was antagonised by adenosine A2 receptor antagonist, 8 sulfophenyltheophylline (8-SPT, 50 x 10(-6) M). The antagonism by 8-SPT was lower in SHR as compared to WKY tissues. L-monomethylarginine (L-LMMA) (30 x 10(-6) M) significantly shifted the CAD relaxation to the right, which was reversed by the addition of L-arginine (100 x 10(-6) M) in both SHR and WKY rats. However, the rightward shift by L-NMMA was smaller in SHR compared to WKY vascular tissues. Vasorelaxation response to acetylcholine (1 x 10(-6) M) was significantly inhibited (50%) in SHR rings compared to WKY. The relaxation produced by sodium nitroprusside (10(-9)-10(-5) M) in endothelium-intact and -denuded aortic rings showed no difference between SHR and WKY. Isoproterenol produced concentration dependent (10-9-10-5 M) relaxation, which was shifted to the right in SHR compared to WKY rings with an intact endothelium, while the removal of endothelium abolished the difference in the response between SHR and WKY. The results suggest: (i) adenosine A2 receptors mediate vasorelaxation in part through endothelium possibly by releasing nitric oxide (NO); (ii) the impairment of endothelium may be one of the factors for the attenuation of adenosine receptor and receptor-mediated responses in SHR. PMID- 11903500 TI - Vanadium pharmacokinetics and oral bioavailability upon single-dose administration of vanadyl sulfate to rats. AB - Vanadium pharmacokinetic parameters and oral bioavailability were determined after administration of vanadyl sulfate, an antidiabetic agent, to male Wistar rats. An optimal sampling design was used over a 21-day period; vanadium was measured in blood by atomic absorption spectrophotometry (AAS). After i.v. bolus injection (3.025 mg V/kg body weight), a three-compartment model was fitted to the data. Mean (+/- SD) half-lives were 0.90 +/- 0.56 hours, 24.8 +/- 14.5 h and 201 +/- 74 h, respectively, for the three phases observed. Vanadium clearance averaged 37.6 +/- 15.8 mL/h. Initial volume of distribution was 2.43 +/- 1.22 L/kg whereas total volume of distribution was 25.4 +/- 3.9 L/kg; these values largely exceeded body weight (i.e. 300 g), in agreement with a great uptake and retention of vanadium in tissues. After oral gavage administration (15.12 and 7.56 mg V/kg body weight), vanadium disposition was best described by a three compartment model, with absorption appearing to occur by a zero-order rate. This process lasted 10.3 +/- 1.3 h and 10.9 +/- 1.1 h for the two dosage levels, respectively. Half-lives corresponding to the terminal log-linear part of the curve were 173.5 +/- 1.6 h and 172 +/- 6 h (Bayesian estimates). No dose dependency was observed for any of the parameters determined. Absolute bioavailabilities, with reference to the i.v. administration, were 12.5% and 16.8% when determined from AUCmod. Bioavailability appeared to be higher than generally stated in the literature. PMID- 11903502 TI - Mechanisms of decreased bradykinin- induced vasodilation in experimental hyperlipemia-hyperglycemia: contribution of nitric oxide and Ca2+-activated K+ channels. AB - Common complications of diabetes are accelerated atherosclerosis and vascular disturbances. We investigated whether the simultaneous insult of hyperlipemia hyperglycemia affects the reactivity of the resistance arteries to bradykinin (BK), and if so, what are the mechanisms responsible for this disturbance. Experiments were conducted on male Golden Syrian hamsters rendered hyperlipemic (H) by a fat-rich diet, diabetic (D) by streptozotocin injection, or simultaneously hyperlipemic-diabetic (HD). Normal age-matched animals were used as controls (C). At 24 weeks after the induction of disease(s) the vascular reactivity of the mesenteric resistance arteries to BK (10(-8)-10(-4) M) was assayed by the myograph technique. To explore the role of nitric oxide (NO) in modulating the endothelium-dependent BK-induced relaxation, two experimental approaches were employed: (i) in vivo administration of L-arginine (622.14 mg/kg bw) to H, D, and HD hamsters (for 12 weeks); (ii) in vitro blockage of nitric oxide synthase by N(omega)-nitro- L-arginine methyl ester (10(-4) M). To evaluate the contribution of Ca2+-activated K+ channel(s) to BK-induced relaxation, the resistance arteries were exposed to 10(-3) M tetraethylammonium. Comparatively, the endothelium-independent relaxation was assayed using sodium nitroprusside (10(-8)-10(-4) M). The results showed that compared to the H and D groups, the HD hamsters exhibited the most reduced vasodilation of the resistance arteries to BK (34.09 +/- 1.5%). The diminished vasodilation was found to be due to a dual mechanism: an L-arginine:NO pathway and a NO-independent process, mediated via Ca2+-activated K+ channels. In vivo administration of L-arginine had favourable effects especially in the HD group, which manifested (i) an; 30% improvement of attenuated BK relaxation, (ii) an increase in sensitivity of the response to BK, (iii) a 3-fold diminishment of plasma hyperglycemia. Collectively, these data explain in part, the mechanisms and possible ways to correct the arterial endothelial dysfunction when diabetes is complicated with hyperlipemia. PMID- 11903503 TI - Effects of some divalent cations on nitrergic relaxations in the mouse corpus cavernosum. AB - Acute effects of some divalent cations (Cd2+, Ni2+, Co2+, Zn2+, Mn2+ and Sn2+) were investigated on neurogenic and endothelium-dependent relaxations in the isolated mouse corpus cavernosum. Neither neurogenic nor endothelium-dependent relaxation was affected by cations at the concentrations used (up to 100 microM), except Cd2+. Although Cd2+ (20 and 40 microM) did not cause any significant alteration in the acetylcholine- (ACh) or sodium nitroprusside- (SNP) induced relaxation, it inhibited electrical field stimulation- (EFS) produced relaxation significantly. Zn2+ and selenium could not reverse this inhibitory action. Cd2+ did block the EFS-evoked guanethidine-sensitive contraction in the presence of N(G)-nitro-L-arginine. Elevation of external Ca2+ content significantly reduced the inhibitions due to Cd2+ on the EFS-induced relaxation and on the EFS-evoked guanethidine-sensitive contraction. In the Ca2+-omitted medium, EFS-induced relaxation disappeared, while acetylcholine-elicited relaxation resisted. Verapamil was ineffective on the relaxation produced by EFS or acetylcholine. However, it significantly diminished phenylephrine-induced contractions. These findings suggest that unlike other cations at the concentrations used in the present study, Cd2+ may have an effect on an external Ca2+-dependent mechanism at the neuronal level, and this effect may be responsible for its acute inhibitory action on the neurogenic relaxation in the mouse corpus cavernosum. PMID- 11903504 TI - Cooling-induced gastrointestinal smooth muscle contractions in the rat. AB - The aim of this study was to assess the effect of cooling on smooth muscle contraction in various parts of the gastrointestinal tract (esophagus, stomach, duodenum, jejunum and colon) and to investigate the basic mechanism underlying cooling-induced (CIC) tonic and rhythmic contractions. Recordings of isometric tension from smooth muscle strips of different parts of the rat gastrointestinal tract were performed using organ-bath techniques, and stepwise cooling was applied. Cooling was tested before and after the addition of various standard agents interfering with known neurogenic (autonomic blockers, tetrodotoxin, capsaicin) and myogenic mechanisms of contraction (calcium channel blockers, Sarcoplasmatic and Ca2+-ATPase pump inhibitors). Step-wise cooling (37 degrees C to 5 degrees C) of all gastrointestinal smooth muscle preparations induced reproducible graded tonic contractions, inversely proportional to temperature. CIC was most pronounced in the jejunum. Cooling abolished rhythmic smooth muscle activity. CIC was not dependent on a neural mechanism nor the release of neurotransmitters, but linked to translocation of calcium. It was reduced by incubation in Ca2+-free solution. Blockage of the Ca2+-ATPase pump, which inhibits the extrusion of calcium, plays a significant role in the process and enhances CIC. Cooling of gastrointestinal smooth muscle preparations induces graded myogenic contractions inversely proportional to the temperature. The mechanism is not dependent on local nervous control but related to a temperature sensitive process of calcium translocation. PMID- 11903506 TI - Sigma receptors: from discovery to highlights of their implications in the cardiovascular system. AB - Sigma receptors are the targets of many ligands, of which some (the haloperidol for instance) are psychoactive, and of substances known to have antiarrhythmic effects (amiodarone and clofilium). They are involved in a variety of cardiovascular functions, such as the regulation of cardiac contractility and rhythm and the regulation of coronary and peripheral arterial vasomotricity. This short review will focus on some aspects regarding the ligands, the binding sites, the intracellular coupling and the cardiovascular functions of these enigmatic receptors. PMID- 11903505 TI - Isoniazid pharmacokinetics in children according to acetylator phenotype. AB - The pharmacokinetics of isoniazid (INH) was studied in children (0-196 months old) according to their acetylator phenotype, estimated from the metabolic acetyl INH/INH molar plasma concentration ratio (MR) measured 3 h after INH oral administration. There were 17 slow (MR < 0.48) and 17 fast acetylators (MR > or = 0.48). The mean apparent plasma clearance was significantly lower, the mean apparent volume of distribution higher and the half-life longer in the slow acetylator group (C1, 0.298 +/- 0.099 L/h/kg; Vd, 1.56 +/- 0.65 L/kg; t1/2, 3.88 +/- 01.89 h) than in the fast acetylator group (Cl, 0.528 +/- 0.234 L/h/kg; Vd, 1.06 +/- 0.45; t1/2, 1.64 +/- 1.1 h). The half-life decreased with age. An impaired isoniazid elimination was suggested in children less than three months old, which may be in favour of an individual dose adjustment in this population. PMID- 11903507 TI - Vascular reactivity to angiotensin II alone or combined with a thromboxane A2 mimetic in the isolated perfused kidney of Lyon hypertensive rats. AB - The aim of this study was to evaluate whether thromboxane A2-prostaglandin H2 (TP) receptor activation potentiates the renal vasoconstrictor effect of Angiotensin II (Ang II) in genetically hypertensive rats of the Lyon strain (LH). Concentration-response curves (CRCs) to Ang II (5 pM to 10 nM), to the specific TP receptor agonist U46619 (7.5-960 nM) and to a mixture of Ang II + U46619 (fixed molar ratio of 1 : 9) were obtained in single-pass perfused kidneys isolated from 8 week-old LH and low blood pressure (LL) control rats. Baseline vascular resistance was significantly increased in LH compared to LL kidneys. Comparison of the CRCs obtained for Ang II and U46619 showed that, in both strains, Ang II was about 100 times more potent than U46619. For both drugs, the pD2 or slope values did not differ among the two strains. Co-activation of TP receptors, analyzed with the method of Poch and Holzmann, tended to potentiate the effects of Ang II in LH but not in LL kidneys. In conclusion, isolated perfused kidneys of LH rat did not exhibit an increased vascular sensitivity to acute infusion of Ang II or U46619 compared to control LL ones. In addition, the results suggest that the interactions between Ang II and TP receptor agonist may differ among the two strains. PMID- 11903508 TI - Non-contribution of renin-angiotensin system to pressor response to N(G)-nitro-L arginine in dogs. AB - Acute systemic blockade of nitric oxide (NO) production by nonselective inhibitors of NO synthase (NOS) isoforms, including N(G)-nitro-L-arginine methyl ester (L-NAME) and N(G)-nitro-L-arginine (L-NNA), has been shown to produce a long-lasting pressor response in conscious and anaesthetised animals. The present study was undertaken to clarify whether the renin-angiotensin system contributes to the development of this pressor response to L-NNA. Systemic blood pressure and heart rate were continuously monitored in dogs anaesthetised with pentobarbital. Plasma renin activity in the blood obtained from a femoral artery and a renal vein was measured by use of radioimmunoassay. The acute pressor response produced by the intravenous administration of L-NNA was accompanied by reduced renin activity in both systemic and renal vascular beds. Captopril, an angiotensin converting enzyme inhibitor, counteracted the pressor response to L-NNA, whereas candesartan, an angiotensin AT1-receptor antagonist, had no apparent effect on it. The counteraction by captopril of the L-NNA-induced pressor response was likely to be attributable to enhancement by captopril of depressor responses to bradykinin, as HOE-140, a bradykinin B2 receptor antagonist, neutralised the effect of captopril. These results suggest that the pressor response acutely produced by the intravenous injection of a NOS inhibitor is not mediated by the renin-angiotensin system in anaesthetised dogs. PMID- 11903509 TI - Effects of tramadol on behavioural indicators of colic pain in a rat model of ureteral calculosis. AB - This study investigated the effect of prolonged administration of tramadol vs. placebo on behavioural indicators of ureteral pain and referred lumbar muscle hyperalgesia in a rat model of artificial ureteral calculosis. Four groups of 10 rats each (female, Sprague-Dawley) were treated twice a day, for 4 days, with i.p. injections of tramadol 1.25 mg/kg, 2.5 mg/kg, 5 mg/kg or saline, respectively. The first injection was delivered 45 min before laparotomy (under pentobarbital anaesthesia) for formation of the stone in the upper left ureter via injection of dental cement. All rats were video-taped 24 h non-stop from the immediate postoperative period until the 4th day for recording of behavioural ureteral crises indicative of colic pain. Lumbar muscle sensitivity was tested daily over the same period by verifying presence or absence of vocalization upon pinching of the parietal layers at L1 level, bilaterally, at a constant predefined pressure value with calibrated forceps. Tramadol significantly reduced number and global duration (ANOVA, P < 0.008 and P < 0.004) of ureteral crises with respect to saline and the effect was dose-dependent (linear regression analysis between doses and parameters of crises, P < 0.003 and P < 0.002). The drug also significantly reduced the incidence of referred muscle hyperalgesia (ANOVA, P < 0.0001). It is concluded that tramadol is highly effective in controlling pain phenomena from urinary stones and can represent a valid therapeutic approach in patients with urinary colics. PMID- 11903510 TI - Interaction of chelerythrine with inositol phosphate metabolism. AB - Chelerythrine, a potent inhibitor of protein kinase C (PKC), was evaluated for its effect on inositol phosphate (IP) metabolism in newborn rat cardiomyocytes in culture. In a first step, we evaluated the effect of chelerythrine on IP accumulation in basal conditions. For a 10(-4) M dose, 5-phosphatase activity (which dephosphorylates IP3 into IP2) was completely blocked and we observed a large increase in IP accumulation limited to IP2 without any increase in IP3, strongly suggesting that chelerythrine at this dose modifies IP metabolism. At a lower dose (10(-5) M) of chelerythrine, which did not modify IP accumulation and 5-phosphatase activity in basal conditions, the response to angiotensin II stimulation was completely abolished by the addition of chelerythrine. We conclude thus that chelerythrine, even at 10(-5) M, interacts markedly with IP metabolism, and caution should be exerted when interpreting the results obtained with this drug, which is still currently used at this dose. PMID- 11903511 TI - 5-fluorocytosine-related bone-marrow depression and conversion to fluorouracil: a pilot study. AB - The aim of this study is to investigate whether fluorouracil (5-FU) could be responsible for bone-marrow depression occurring in fluorocytosine (5-FC) treated patients. Six 5-FC treated patients were included in this pilot study. Toxicity was monitored by means of thrombocyte and leucocyte counts. 5-FC and 5-FU serum levels were measured using a high-performance liquid chromatography (HPLC) assay that allows simultaneous determination of both compounds. The amounts of 5-FU in the 34 available serum samples remained below the limit of quantitation (< 0.05 mg/L), whereas 5-FC levels could be detected in all samples. Instead, low levels of the 5-FU catabolite alpha-fluoro-beta-alanine (FBAL) were detected in several of the investigated serum samples. In case of three patients thrombocyte counts remained within the normal range during 5-FC treatment, whereas one patient developed thrombocytopenia (50 x 10(9) thrombocytes/L) during therapy. Furthermore, one patient developed leucocytopenia (2.6 x 10(9) leucocytes/L) during 5-FC therapy, whereas the remaining five patients were suffering from leucocytosis prior to 5-FC therapy. In conclusion, we found nondetectable 5-FU serum concentrations (< 0.05 mg/L) in ICU patients treated with intravenous 5-FC, making it unlikely that 5-FC-associated toxicity results from 5-FU exposure in patients receiving intravenous 5-FC therapy. These findings may be explained by the fact that our patients received 5-FC intravenously instead of orally, therefore not allowing active conversion of 5-FC to 5-FU by the human intestinal microflora. PMID- 11903512 TI - Adverse drug reactions: keeping up to date. AB - The amount of published literature on adverse drug reactions is overwhelming; for example, the serial publication Side Effects of Drugs Annual lists and critically discusses over 3000 references each year. As a group, pharmacotherapeutics journals publish more papers on adverse drug reactions than journals that cover other fields, but even so they publish a minority of the total number of papers, and no single journal or group of journals can be highlighted as being a frequent source of primary information. Non-specialists must therefore rely on secondary literature (reviews) and tertiary literature (critical summaries) for information. Most of the primary published literature is in the form of anecdotal reports (30%) and formal studies or randomized controlled trials (35%). The anecdotal reports vary in quality; a new serial publication devoted to this type of article would bring some of the literature together and would improve the quality of reporting. Although many of the randomized controlled trials are of good quality and large enough to reveal benefit, most are too small to provide robust information about adverse drug reactions. Systematic reviews are too few in number (only 1.25% of publications on adverse drug reactions cited in Side Effects of Drugs Annual); more are needed. PMID- 11903517 TI - The scientist's perspective. PMID- 11903513 TI - Psychomotor and cognitive effects of piribedil, a dopamine agonist, in young healthy volunteers. AB - Piribedil is a dopamine agonist acting on D2 and D3 central nervous system dopamine receptors. This drug has been administered to 12 young healthy male volunteers (age 22 +/- 2 years) according to a single center randomized, double blind, two ways cross-over, placebo controlled trial, including a washout period of one week. Placebo and piribedil were administered by a single intravenous infusion over 2 h (3 mg). Psychomotor performance and cognitive functions were assessed through a standardized and computerized psychometric tests battery and a continuous electroencephalogram (EEG) mapping. Piribedil improved simple reaction time (P=0.02), immediate (P=0.045 and 0.004), and delayed free recall (P=0.05), dual coding test (P=0.02) and increased theta and fast beta waves on the EEG (P < 0.05 and 0.001, respectively). No deleterious effect was observed on the tests exploring attention and concentration via the other procedures. It is concluded that a single intravenous perfusion of piribedil 3 mg improves alertness and the information processing speed within the central nervous system, in healthy volunteers. PMID- 11903518 TI - The clinician's perspective. PMID- 11903519 TI - The industry's perspective partnership: advancing the care of persons with headache. PMID- 11903520 TI - Managing migraine: a patient's perspective. PMID- 11903522 TI - Images from Headache. PMID- 11903523 TI - Butalbital in the treatment of headache: history, pharmacology, and efficacy. AB - Analgesics containing butalbital compounded with aspirin, acetaminophen, and/or caffeine are widely used for the treatment of migraine and tension-type headache. The butalbital-containing compounds are efficacious in placebo-controlled trials among patients with episodic tension-type headaches. Despite their frequent clinical use for migraine, they have not been studied in placebo-controlled trials among patients with migraine. Barbiturates can produce intoxication, hangover, tolerance, dependence, and toxicity. Butalbital can result in intoxication that is clinically indistinguishable from that produced by alcohol. Butalbital-containing analgesics can produce drug-induced headache in addition to tolerance and dependence. Higher doses can produce withdrawal syndromes after discontinuation. Butalbital-containing analgesics may be effective as backup medications or when other medications are ineffective or cannot be used. Because of concerns about overuse, medication-overuse headache, and withdrawal, their use should be limited and carefully monitored. PMID- 11903525 TI - Comparison of intravenous valproate versus intramuscular dihydroergotamine and metoclopramide for acute treatment of migraine headache. AB - OBJECTIVE: To determine the effectiveness and tolerability of intravenous valproate for the acute treatment of migraine headache with or without aura (International Headache Society diagnostic criteria 1.1 and 1.2) compared with intramuscular metoclopramide 10 mg followed 10 minutes later by intramuscular dihydroergotamine 1 mg. BACKGROUND: Divalproex sodium is approved for prophylaxis of migraine headache. We studied the possible effectiveness of intravenous sodium valproate for the treatment of acute migraine headache. Valproate offers a treatment option for patients with migraine who recently have used a triptan or dihydroergotamine, theoretically avoiding the risk of drug interactions or cardiovascular complications. DESIGN/METHODS: In an open-label randomization, patients with an established diagnosis of migraine with or without aura were administered either intravenous valproate or intramuscular dihydroergotamine with metoclopramide to treat moderate-to-severe migraine headache of 24 to 96 hours' duration. Forty patients alternately received either 500 mg intravenous valproate or 10 mg metoclopramide intramuscularly followed by 1 mg dihydro- ergotamine. Patients rated severity of headache and the presence or absence of nausea, photophobia, or phonophobia at baseline, and at 1, 2, 4, and 24 hours. RESULTS: With intravenous valproate, 50% of patients reported headache improvement from moderate or severe to none or mild at 1 hour following treatment, 60% reported such improvement at 2 hours, 60% at 4 hours, and 60% at 24 hours. Corresponding improvement rates for dihydroergotamine were 45% at 1 hour, 50% at 2 hours, 60% at 4 hours, and 90% at 24 hours. Intravenous valproate and intramuscular dihydroergotamine provided similar relief from associated migrainous symptoms (nausea, photophobia, and phonophobia) during the first 4 hours following treatment. While none of the patients who received intravenous valproate experienced drug-related side effects during treatment, 15% of patients who took dihydroergotamine experienced one or more episodes of nausea and diarrhea during the first 4 hours of treatment. CONCLUSIONS: Intravenous valproate is similar in effectiveness to dihydroergotamine/metoclopramide as abortive therapy for prolonged moderate-to-severe acute migraine headache. Although the results were not statistically significant (P =.3635), intravenous valproate appears to offer a safe, effective, and well-tolerated treatment for patients with acute migraine. Relative to dihydroergotamine/metoclopramide, however, headache relief was not as likely to be sustained at 24 hours as with intravenous valproate. PMID- 11903524 TI - Topiramate in migraine prevention: a double-blind, placebo-controlled study. AB - OBJECTIVE: To evaluate the efficacy of topiramate in the preventative treatment of episodic migraine. BACKGROUND: Topiramate is a broad-spectrum antiepileptic drug effective for treatment of multiple seizure types in adults and children. Antiepileptic agents have demonstrated efficacy in migraine prevention, and open label experience from our clinic has suggested that topiramate might be effective for this use. We consequently conducted a single-center, double-blind, placebo controlled trial to evaluate the efficacy and safety of topiramate for the preventative treatment of migraine. METHODS: Forty patients, aged 19 to 62 years (mean, 38.2 years), were randomly assigned in a 1:1 ratio to receive topiramate (n = 19; all women) or placebo (n = 21; 20 women, 1 man). Following a prospective baseline phase of 4 weeks, the study drug dose was titrated weekly in 25-mg increments over 8 weeks to 200 mg per day or to the maximum tolerated dose. The titration phase was followed by an 8-week maintenance phase. RESULTS: During the entire double-blind phase, topiramate-treated patients experienced a significantly lower 28-day migraine frequency (3.31 +/- 1.7 versus 3.83 +/- 2.1; P =.002) compared to placebo, irrespective of use of concomitant migraine prevention medications. The mean 28-day migraine frequency was reduced by 36% in patients receiving topiramate as compared with 14% in patients receiving placebo (P =.004). Twenty-six percent of the patients on topiramate and 9.5% of the patients on placebo achieved a 50% reduction in migraine frequency (P >.05). The mean dose of topiramate was 125 mg per day (range, 25 to 200 mg per day). Topiramate was well tolerated; 2 of 19 topiramate-treated patients discontinued treatment due to adverse events. Adverse effects that occurred more frequently in topiramate-treated patients included paresthesia, weight loss, altered taste, anorexia, and memory impairment. CONCLUSIONS: Preventative therapy with topiramate significantly reduced migraine frequency. Larger multicenter clinical studies may further delineate the role of topiramate in migraine prevention. PMID- 11903526 TI - Triptans in the treatment of basilar migraine and migraine with prolonged aura. AB - OBJECTIVE: To report on the use of triptans in migraine with prominent neurologic symptoms. BACKGROUND: As stated in their package inserts, the triptans are contraindicated in patients with basilar or familial hemiplegic migraine, and physicians are reluctant to prescribe these drugs to other patients with prominent or prolonged aura. METHODS: We evaluated 13 patients with basilar migraine, familial hemiplegic migraine, or migraine with prominent or prolonged aura who had received triptans. RESULTS: Excellent; no adverse events. CONCLUSION: The contraindication of triptans in basilar migraine should be reconsidered. Similarly, prominent or prolonged aura may not represent a reasonable contraindication to triptan therapy. PMID- 11903527 TI - Botulinum toxin type A and EMG: a key to the understanding of chronic tension type headaches? AB - BACKGROUND: The pathogenesis of chronic tension-type headache remains unclear, and the role of muscle tension is especially controversial. Botulinum toxin type A, a potent inhibitor of muscle tone, has been used to treat chronic tension-type headache. OBJECTIVE: To determine whether clinical response to treatment of chronic tension-type headache with Botox A parallels changes in resting muscle activity recorded through serial electromyography (EMG). METHODS: We randomly assigned eight patients with chronic tension-type headache to pericranial injection of 500 MU Botox A versus placebo (isotonic saline). RESULTS: At 6 and 12 weeks following treatment, there were no significant differences in clinical outcome between the placebo and the Botox A groups. This occurred despite EMG evidence of a reduction in resting muscle activity in the Botox A-treated patients. CONCLUSION: These results support the hypothesis that peripheral mechanisms such as increased muscle tone play, at most, a minor role in the pathophysiology of chronic tension-type headache. PMID- 11903528 TI - Occipital nerve blocks and managed care: a review of the reviewers. PMID- 11903529 TI - Lower cervical intramuscular injections for headache relief. PMID- 11903534 TI - Antiepileptic drugs: how they work in headache. AB - Antiepileptic drugs (AEDs) are promising agents for the prevention of migraine and other head pain. Migraine and epilepsy share several clinical features and respond to many of the same pharmacologic agents, suggesting that similar mechanisms may be involved in their pathophysiology. The mechanisms of action of AEDs are not fully understood, and a single drug may have more than one mechanism, both in epilepsy and in migraine. Valproate, topiramate, and gabapentin are likely to affect nociception by modulating gamma-aminobutyric acid (GABA-) and/or glutamate-mediated neurotransmission. All three AEDs enhance GABA mediated inhibition. Valproate and gabapentin interfere with GABA metabolism to prevent its ultimate conversion to succinate, and topiramate potentiates GABA mediated inhibition by facilitating the action of GABA receptors. In addition, topiramate acts directly on non-N-methyl-D-aspartate, alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid/kainate glutamate receptors. Valproate, topiramate, and possibly gabapentin inhibit sodium ion channels. All three drugs modulate calcium ion channel activity. Valproate blocks T-type calcium ion channels; topiramate inhibits high-voltage-activated L-type calcium ion channels; and gabapentin binds to the alpha2delta subunit of L-type calcium ion channels. AEDs may be useful in migraine prevention through such mechanisms as modulating the biochemical phenomena of aura or acting directly on the nociceptive system. Further evaluations of AEDs in migraine models will provide a better understanding of the pathophysiology and prevention of migraine. PMID- 11903535 TI - Shared mechanisms and comorbidities in neurologic and psychiatric disorders. AB - Migraine may be comorbid with several other neurologic and psychiatric conditions, including mood disorders (eg, depression, anxiety, panic disorder), epilepsy, stroke, and essential tremor. Comorbidity presents physicians with opportunities and challenges for both diagnosis and treatment. All diseases must be considered, and therapeutic strategies may need to be modified to avoid potential drug interactions. Comorbidities also may provide clues to the pathophysiologies and any shared mechanisms of the two disorders. Longitudinal studies have demonstrated a bidirectional influence between migraine and major depression, but not between migraine and other severe headache. Migraine is strongly and consistently associated with panic disorder. The risk of migraine in epilepsy is increased particularly in individuals with head trauma, partial seizures, and a positive family history of migraine. The influence is bidirectional. There is also growing evidence of an association between migraine and stroke, particularly among women of childbearing age and individuals who experience migraine with aura. Lastly, a bidirectional association between migraine and essential tremor also exists. These findings suggest that migraine, major depression, epilepsy, and essential tremor share one or more common etiologies. Clinicians should be mindful of them as they design treatment strategies, and also should consider the use of a single pharmacologic agent that is effective for all conditions. PMID- 11903536 TI - Antiepileptic drugs in migraine prevention. AB - Migraineurs may continue to experience attacks, despite daily use of one or more agents from a wide range of drugs, including beta-blockers, calcium channel blockers, serotonin antagonists, tricyclic antidepressants, monoamine oxidase inhibitors, and antiepileptic agents. Divalproex sodium is the only antiepileptic drug approved for migraine prevention. Gabapentin, topiramate, and other antiepileptic agents are being evaluated for migraine prevention and treatment. Prospective, double-blind, placebo-controlled clinical trials of divalproex, gabapentin, and topiramate for migraine prevention generally were composed of a prospective baseline period, a dose titration period, and a fixed-dose treatment period. The primary efficacy variable was a reduction in the 28-day frequency of migraine headache. Patients receiving divalproex for 12 weeks at doses up to 1500 mg/day achieved significant decreases in the migraine frequency (P<.05), corresponding to reductions of 30% to 40% compared with baseline. Nearly half of the divalproex-treated patients had a 50% or more reduction from baseline in headache frequencies (P< or =.05). Asthenia, vomiting, somnolence, tremor, and alopecia were common adverse events associated with divalproex. Significant reductions in migraine frequency were also observed with gabapentin (1800 to 2400 mg/day) when compared with placebo (P<.01), and nearly half of all patients treated at the highest dose experienced a reduction in headache rate of 50% or more. Somnolence was the most commonly reported adverse event among the gabapentin-treated patients. Two single-center, double-blind, placebo-controlled clinical trials evaluated topiramate for migraine prevention. A lower 28-day migraine frequency was seen during 18 weeks of administration at a maximum daily dose of 200 mg (P =.09). In a second study, a significantly lower mean 28-day migraine frequency was observed during 16 weeks of treatment with topiramate (P =.0015). Mean reduction in migraine frequency was also significantly greater in topiramate-treated patients (P =.0037). Paresthesias, diarrhea, somnolence, and altered taste were commonly reported adverse events in the topiramate-treated patients. Unlike some patients given divalproex or gabapentin, some given topiramate reported weight loss. Large, double-blind, placebo-controlled trials may prove the effectiveness of novel antiepileptic drugs in migraine prevention. PMID- 11903537 TI - Antiepileptic drugs in the management of cluster headache and trigeminal neuralgia. AB - Cluster headache and trigeminal neuralgia are relatively rare but debilitating neurologic conditions. Although they are clinically and diagnostically distinct from migraine, many of the same pharmacologic agents are used in their management. For many patients, the attacks are so frequent and severe that abortive therapy is often ineffective; therefore, chronic preventive therapy is necessary for adequate pain control. Cluster headache and trigeminal neuralgia have several distinguishing clinical features. Cluster headache is predominantly a male disorder; trigeminal neuralgia is more prevalent in women. Individuals with cluster headaches often develop their first attack before age 25; most patients with trigeminal neuralgia are between age 50 and 70. Cluster headaches are strongly associated with tobacco smoking and triggered by alcohol consumption; trigeminal neuralgia can be triggered by such stimuli as shaving and toothbrushing. Although the pain in both disorders is excruciating, cluster headache pain is episodic and unilateral, typically surrounds the eye, and lasts 15 to 180 minutes; the pain of trigeminal neuralgia lasts just seconds and is usually limited to the tissues overlying the maxillary and mandibular divisions of the trigeminal nerve. Cluster headache is unique because of its associated autonomic symptoms. Although the pathophysiology of cluster headache and trigeminal neuralgia are not completely understood, both appear to have central primary processes, and these findings have prompted investigations of the effectiveness of the newer antiepileptic drugs for cluster headache prevention and for the treatment of trigeminal neuralgia. The traditional antiepileptic drugs phenytoin and carbamazepine have been used for the treatment of trigeminal neuralgia for a number of years, and while they are effective, they can sometimes cause central nervous system effects such as drowsiness, ataxia, somnolence, and diplopia. Reports of studies in small numbers of patients or individual case studies indicate that the newer antiepileptic drugs are effective in providing pain relief for trigeminal neuralgia and cluster headache sufferers, with fewer central nervous system side effects. Divalproex has been shown to provide effective pain control and to reduce cluster headache frequency by more than half in episodic and chronic cluster headache sufferers. Topiramate demonstrated efficacy in a study of 15 patients, with a mean time to induction of cluster headache remission of 1.4 weeks (range, 1 day to 3 weeks). In the treatment of trigeminal neuralgia, gabapentin has been shown to be effective in an open-label study. When added to an existing but ineffective regimen of carbamazepine or phenytoin, lamotrigine provided improved pain relief; it also may work as monotherapy. Topiramate provided a sustained analgesic effect when administered to patients with trigeminal neuralgia. The newer antiepileptic drugs show considerable promise in the management of cluster headache and trigeminal neuralgia. PMID- 11903539 TI - Low-dose tizanidine with nonsteroidal anti-inflammatory drugs for detoxification from analgesic rebound headache. AB - OBJECTIVE: To describe an outpatient regimen for analgesic detoxification and resolution of analgesic rebound headache. BACKGROUND: Frequent analgesic use is believed to promote the transformation of episodic migraine into a chronic, pervasive headache syndrome. Management of pain precipitated by analgesic withdrawal is crucial to treatment success. Outpatient treatment protocols designed to achieve successful withdrawal will reduce costs and potentially lead to more widespread implementation of therapy. METHODS: Patients with appropriate histories were managed on an outpatient basis for detoxification by discontinuation of the offending analgesic and initiation of treatment with tizanidine and a long-acting nonsteroidal anti-inflammatory drug. Patients kept diaries of pain and medication use. Results were evaluated at 6 and 12 weeks. Patients able to tolerate no or trivial analgesic use (ie, 4 or fewer doses in each 2-week period) were considered responders. RESULTS: At 6 weeks, 36 patients (65%) were responders. At 12 weeks, 38 patients (69%) were responders. The chronic daily headache pattern had resolved at 12 weeks in 34 patients (62%). CONCLUSIONS: This treatment protocol was well tolerated and yielded a high degree of efficacy, demonstrating that outpatient management can be effective for achieving analgesic withdrawal and resolution of analgesic rebound headache. PMID- 11903541 TI - Prevalence of menstrually related migraine and nonmigraine primary headache in female students of Belgrade University. AB - OBJECTIVES: To determine prevalence and characteristics of menstrually related migraine and nonmigraine headache in female students of Belgrade University. METHODS: A questionnaire was administered to female students during randomly selected classes of the Schools of Medicine and Pharmacy. Diagnoses were assigned according to the criteria of the International Headache Society and MacGregor's stricter definition of "menstrual" migraine. RESULTS: Of 1943 female students (18 to 28 years old), 1298 (66.8%) had primary headaches. Among 1298 students with headache, 245 (12.6%) had migraine and 1053 (54.2%) had nonmigraine headache. The prevalence rates of migraine versus nonmigraine headache in relation to the menstrual cycle were: premenstrual, 0.9% versus 4.4%; menstrual, 1.5% versus 1.5%; menstrually associated, 6.1% versus 10.1%; menstrually unchanged, 2.7% versus 19.2%; and menstrually unrelated, 1.4% versus 18.9%. Female students with migraine had menstrually related attacks more frequently than students with nonmigraine headache (67.7% versus 29.5%). This difference was most prominent among students with menstrual migraine compared with students with menstrual nonmigraine headache (12.2% versus 2.7%). Exacerbation of migraine during menstruation was slightly more severe and more complex than exacerbation of nonmigraine headache. Female students with migraine versus nonmigraine headache did not differ significantly in age, age at onset of menarche, or age at onset of headache. Female students with migraine were significantly more likely to report a positive family history for migraine and menstrual migraine, severe attacks, reduced work activity, and aura. CONCLUSION: The results obtained are in accord with the prevailing opinion that there is a relationship between migraine and female sex hormones, and suggest that women with nonmigraine headache are also susceptible to hormonal fluctuations. PMID- 11903540 TI - Coping style and social support in men and women suffering from cluster headache or migraine. AB - OBJECTIVE: To investigate similarities and differences between patients with cluster headache and patients with migraine. BACKGROUND: Patients with migraine and patients with cluster headache are considered, by many clinical neurologists, to be different psychologically and socially. METHODS: Twenty-five age-matched pairs of men and 24 age-matched pairs of women with either migraine or episodic cluster headache (men aged 31 to 62 years; mean, 47 years; women aged 23 to 72 years; mean, 44 years) were compared with regard to coping profiles as reflected in two "coping wheels," one for the present situation and one for the future. In addition, availability of attachment and social interaction was assessed by means of the ISSI (Interview Schedule for Social Interaction). RESULTS: Women with cluster headache anticipated fewer activities for themselves than women with migraine, and findings were similar in the male pairs. The men with cluster headache also anticipated significantly fewer activities for themselves in the present and with others in the present and in the future than the men with migraine. There was no significant difference as to emotional loading between the two groups. A tendency to more optimistic anticipation was found in the women with cluster headache. There were highly significant differences between the two groups in the "future" wheel. The group with migraine expected more concrete activities and more activities with their families in the future, and they also described their present situation to involve more activities with others. CONCLUSIONS: Results from the present study differ from those from studies utilizing more conventional questionnaires. In particular, we found that patients with cluster headache have fewer close social contacts than patients with migraine. PMID- 11903542 TI - Depression, automatic thoughts, alexithymia, and assertiveness in patients with tension-type headache. AB - OBJECTIVE: The role of psychological factors related to headache has long been a focus of investigation. The aim of this study was to evaluate depression, automatic thoughts, alexithymia, and assertiveness in persons with tension-type headache and to compare the results with those from healthy controls. METHODS: One hundred five subjects with tension-type headache (according to the criteria of the International Headache Society classification) and 70 controls were studied. The Beck Depression Inventory, Automatic Thoughts Scale, Toronto Alexithymia Scale, and Rathus Assertiveness Schedule were administered to both groups. Sociodemographic variables and headache features were evaluated via a semistructured scale. RESULTS: Compared with healthy controls, the subjects with headache had significantly higher scores on measures of depression, automatic thoughts, and alexithymia and lower scores on assertiveness. Subjects with chronic tension-type headache had higher depression and automatic thoughts scores than those with episodic tension-type headache. CONCLUSIONS: These findings suggested that persons with tension-type headache have high depression scores and also may have difficulty with expression of their emotions. Headache frequency appears to influence the likelihood of coexisting depression. PMID- 11903544 TI - Primary headaches: a convergence hypothesis. AB - After reviewing the historic differentiation between migraine and tension-type headache, the authors note that the similarities between these two types of primary headaches outweigh the differences, and so hypothesize that these headaches share a common pathophysiology. The convergence hypothesis for primary headaches links the clinical features of an evolving headache to current pathophysiological models. The authors suggest that successive symptoms experienced clinically reflect an escalating pathophysiological process, beginning with the premonitory period and progressing into tension-type headache and, if uninterrupted, finally into migraine. The clinical manifestations of other headache types, such as so-called sinus headache or temporomandibular headache, may also be explained by this model. A convergence hypothesis for primary headaches has important implications for earlier recognition, diagnosis, and treatment. PMID- 11903543 TI - Potential validity of conducting research on headache in internet populations. AB - OBJECTIVE: While it is technically feasible to conduct migraine research on the Internet, can one be sure that the subjects who participate actually have the disorder? We examined this issue in 109 sequential subjects in an Internet study of migraineurs' symptoms. BACKGROUND: The increasing use of the Internet by the US population provides an opportunity for using this medium in clinical research, but, to be useful, the results of such research must be valid and generalizable. METHODS: Using postings to Internet newsgroups of migraineurs, we recruited subjects to a migraine research site. We examined reported symptoms by comparing them with the International Headache Society's criteria for diagnosis of migraine and by subjective review by a neurologist. We also attempted to contact the subject's primary care physicians to confirm their diagnosis. RESULTS: We found considerable evidence for the validity of the participants' self-reported diagnoses. Most subjects with professed migraine reported quality of pain (97%) and associated symptoms (92%) consistent with that diagnosis, and review of their symptoms and questionnaire responses by the neurologist suggested that almost all subjects (97%) had migraine. Personal physicians confirmed a diagnosis of migraine in 90% of consenting subjects (n=49). CONCLUSIONS: The validity of self reported diagnosis of migraine does not appear to be an obstacle to conducting research in subject populations on the Internet. PMID- 11903545 TI - Headache of gastrointestinal origin: case studies. AB - Three patients with headache of gastrointestinal origin are described. Reflux, dyspepsia, and constipation caused the headaches, which responded promptly to treatment of the respective gastrointestinal disorders. PMID- 11903546 TI - Characteristics of headache associated with acute carbon monoxide poisoning. AB - OBJECTIVE: To evaluate systematically the characteristics of headache due to acute exposure to carbon monoxide. BACKGROUND: Headache is the most commonly reported symptom in acute carbon monoxide poisoning. While it is often described as throbbing and diffuse, a systematic characterization of carbon monoxide associated headache has never been published. METHODS: Patients referred for hyperbaric oxygen treatment of acute carbon monoxide poisoning were asked whether headache was part of their symptom complex. When present, specific details about the nature of the headache were collected from 100 consecutive patients through use of a standardized questionnaire. RESULTS: Information on carbon monoxide associated headache was collected from 34 female and 66 male patients with a mean carboxyhemoglobin level of 21.3%+/- 9.3%. The most common location for pain was frontal (66%), although more than one location was involved in 58% of patients. Nature of the pain at any time during its course was dull in 72% of patients and sharp in 36%. Headache was throbbing in 41%, continuous in 74%, and intermittent in 16% of those evaluated. Peak intensity of pain did not correlate with the carboxyhemoglobin level. Headache improved prior to hyperbaric oxygen treatment in 72%, resolving entirely in 21%. Of those with residual headache, pain improved with hyperbaric oxygen in 97%, resolving entirely in 44%. CONCLUSION: The headache accompanying acute carbon monoxide poisoning is extremely variable in nature. "Classic" throbbing, diffuse headaches were rarely described by patients. There are no patterns which can be considered characteristic to aid in the diagnosis of carbon monoxide-induced headache. Similarly, no trait was identified which might allow elimination of carbon monoxide poisoning from the differential diagnosis of headache. PMID- 11903547 TI - Sexual headache and stroke in a heavy cannabis smoker. AB - Sexual headaches usually develop during orgasm. Stroke complicating is rare. We report the case of a young man and heavy cannabis smoker who suffered posterior cerebral artery infarction during his first episode of coital headache. PMID- 11903548 TI - Migraine and Raynaud phenomenon: possible late complications of Kawasaki disease. AB - Migraine and Raynaud phenomenon often coexist and may reflect similar vascular reactions. Both have been associated with vascular endothelial cell dysfunction. Kawasaki disease is a systemic vasculitis of unknown etiology that affects children and may lead to the formation of coronary artery aneurysms. Endothelial cell dysfunction has been demonstrated late in Kawasaki disease and is not restricted to coronary vessels. We report the case of a patient who developed typical migraine with aura and Raynaud phenomenon at the age of 14, 12 years after onset of Kawasaki disease. His migraine responded well to pizotifen, and both migraine and Raynaud phenomenon improved after initiation of treatment with valproic acid. We postulate that both migraine and Raynaud phenomenon in this case represent late consequences of Kawasaki disease and result from extracoronary endothelial dysfunction. PMID- 11903549 TI - Malignant coital headache. PMID- 11903550 TI - Headache and a pupil-sparing third nerve palsy. PMID- 11903551 TI - Physician-assisted suicide and the prescription of narcotics. PMID- 11903554 TI - Re: "Chronic Helicobacter pylori infection and migraine: a case-control study" (Pinessi L, Savi L, Pellicano R, et al. Headache. 2000;40:836-839). PMID- 11903552 TI - Gabapentin in migraine prophylaxis: is it effective and well tolerated? PMID- 11903555 TI - Acetaminophen and metabolite: lack of effect on nitric oxide synthase (constitutive or inducible). PMID- 11903556 TI - What do patterns of genetic variability reveal about mitochondrial recombination? AB - Recent claims that patterns of genetic variability in human mitochondria show evidence for recombination, have provoked considerable argument and much correspondence concerning the quality of the data, the nature of the analyses, and the biological realism of mitochondrial recombination. While the majority of evidence now points towards a lack of effective recombination, at least in humans, the debate has highlighted how difficult the detection of recombination can be in genomes with unusual mutation processes and complex demographic histories. A major difficulty is the lack of consensus about how to measure linkage disequilibrium. I show that measures differ in the way they treat data that are uninformative about recombination, and that when just those pairwise comparisons that are informative about recombination are used, there is agreement between different statistics. In this light, the significant negative correlation between linkage disequilibrium and distance, in at least some of the data sets, is a real pattern that requires explanation. I discuss whether plausible mutational and selective processes can give rise to such a pattern. PMID- 11903557 TI - Haplotype frequency distribution and discriminatory power of two mtDNA fragments in a marine pelagic teleost (Atlantic herring, Clupea harengus). AB - Two regions of the mtDNA of Atlantic herring (Clupea harengus), ND3/4 and ND5/6, were surveyed by RFLP analysis to assess the effects of marker variability on the power of statistical tests. Although haplotype diversity was similar, marked differences in the number of common haplotypes were observed between the two regions, with ND3/4 having a more even distribution of haplotype frequencies. ND3/4 also revealed higher differentiation among samples than ND5/6, similar to previously published microsatellite data of the same samples. The data suggested that the highly skewed haplotype distribution of many mtDNA markers may be one of the reasons for the less frequent detection of population differentiation compared to microsatellites. Pooling ('binning') of related haplotypes did not increase differentiation revealed by each mtDNA region individually, but greatly strengthened geographical patterns shown by both regions combined. The data provided evidence for genetic differentiation of Icelandic herring from other north-eastern Atlantic stocks, and indicated genetic differences between Baltic and Celtic Sea herring. However, mtDNA data failed to confirm previously reported differences between Barents Sea herring, Norwegian fjord populations and Norwegian spring spawners. PMID- 11903558 TI - High levels of diploid male production in a primitively eusocial bee (Hymenoptera: Halictidae). AB - Under single locus complementary sex determination (sl-CSD), diploid males are produced from fertilized eggs that are homozygous at the sex-determining locus. Diploid males are effectively sterile, and thus their production generates a costly genetic load. Using allozyme electrophoresis, a large number of diploid males were detected in natural populations of the primitively eusocial bee, Halictus poeyi Lepeletier collected in southern and central Florida during May 2000. Estimates for the proportion of diploids that are male ranged from 9.1% to 50%, while the frequency of matched matings ranged from 18.2% to 100%. The effective number of alleles at the sex-determining locus ranged from two to 11, with an average of five alleles. The effective population size of Halictus poeyi was estimated to be 19.6 +/- 2.5 SE. These data are interpreted in the light of the biogeographic history of Florida and the social biology/population dynamics of H. poeyi. PMID- 11903559 TI - Molecular and cytogenetic characterization of highly repeated DNA sequences in the vole Microtus cabrerae. AB - The genus Microtus presents several species with extremely large sex chromosomes that contain large blocks of constitutive heterochromatin. Several cytogenetic and molecular studies of the repetitive sequences in species of the genus Microtus have demonstrated that the heterochromatin is highly heterogeneous. We have cloned and characterized a family of repetitive DNA sequences from M. cabrerae, a species with large heterochromatic blocks on the giant sex chromosomes. These repetitive sequences are 65.84% A-T rich, organized in tandem, with a 161-bp unit and are located on the centromeric region of autosomes and the X chromosome. In addition, this repetitive DNA is located throughout the entire heterochromatic block of the X chromosome and on three interstitial bands in the heterochromatic block of the Y chromosome. Comparative analysis of this family of repetitive sequences from three Microtus species revealed that the development of these sequences has occurred by concerted evolution. Our results support the hypothesis that the heterochromatic blocks from the sex chromosomes of different species are evolving independently and they probably have the genetic capacity to amplify and retain different satellite DNAs. For a topic related to the location of these repetitive DNA sequences on the Y chromosome of M. cabrerae, we propose a model to explain the origin of a length polymorphism previously described for this chromosome. PMID- 11903560 TI - Genetic differentiation of the malaria vector Anopheles gambiae across Nigeria suggests that selection limits gene flow. AB - Gene flow was investigated in Anopheles gambiae from eight localities that span the ecological zones of Nigeria (arid savanna zones in the north gradually turn into humid forest zones in the south). Genetic differentiation was measured over 10 microsatellite loci and, to determine any effects of selection, five loci were located within chromosome inversions and the other five were outside inversions. Over all loci, the largest estimates of differentiation were in comparisons between localities in the savanna vs. forest zones (range FST 0.024-0.087, Nm 2.6 10.1; RST 0.014-0.100, Nm 2.2-16.4). However, three loci located within inversions on chromosome II, whose frequencies varied clinically from north to south, were responsible for virtually all of the differentiation. When the three loci were removed, genetic distances across the remaining seven loci were markedly reduced even between localities in the forest and savanna zones (range FST 0.001-0.019, Nm 12.7-226.1) or no longer significant (P > 0.05) in the case of RST. Although tests of isolation by distance gave seemingly equivocal results, geographical distance does not appear to limit gene flow. These observations suggest that gene flow is extensive across the country but that selection on genes located within some inversions on chromosome II counters the homogenizing effects of gene flow. PMID- 11903561 TI - Meiotic pairing of sex chromosome fragments and its relation to atypical transmission of a sex-linked marker in Ephestia kuehniella (Insecta: Lepidoptera). AB - The physical basis of non-Mendelian segregation of a sex-linked marker was studied in sex- chromosome mutant females of eight ASF ('abnormal segregating females') lines in the flour moth, Ephestia kuehniella. Electron microscopical analysis of microspread synaptonemal complexes revealed that in one line, the Z chromosome segment that contained the dz+ allele was translocated onto an autosome. The resulting quadrivalent visible in early female meiosis was 'corrected' into two bivalents in later stages. This explains autosomal inheritance of the sex chromosome marker in this strain. In the other seven ASF lines, the type of meiotic pairing of an additional fragment (Zdz+) of the Z chromosome was responsible for abnormal segregation of the marker gene. In several of these lines, Zdz+ contained a piece of the W chromosome in addition to the Z segment, as was confirmed by comparative genomic hybridization (CGH). Zdz+ formed three alternative pairing configurations with the original sex chromosomes: (i) a WZZdz+ trivalent, (ii) a WZ bivalent and a Zdz+ univalent or (iii) a ZZdz+ bivalent and a W univalent. In the most frequent WZZdz+ configuration, Zdz+ synapsed with Z and, consequently, segregated with W, simulating W linkage. This explains the predominant occurrence of the parental phenotypes in the progeny. Zdz+ univalents or W univalents, on the other hand, segregated randomly, resulting in both parental and nonparental phenotypes. In two of these lines, the Zdz+ was transmitted only to females. The results suggest that the W chromosome segment in Zdz+ of these lines contains a male-killing factor which makes it incompatible with male development. Our data provide direct evidence for the regular transmission of radiation-induced fragments from lepidopteran chromosomes through more than 50 generations. This is facilitated by the holokinetic nature of lepidopteran chromosomes. We conclude that Zdz+ fragments may persist as long as they possess active kinetochore elements. PMID- 11903562 TI - Ribosomal genes in Coregonid fishes (Coregonus lavaretus, C. albula and C. peled) (Salmonidae): single and multiple nucleolus organizer regions. AB - Major rDNA loci, i.e. nucleolus-organizing regions (NORs), were assigned using chromomycin-A3 (CMA3) staining followed by sequential silver (Ag) staining and in situ hybridization (ISH) with a rDNA probe to the chromosomes of the European whitefish (Coregonus lavaretus), the peled (Coregonus peled) and the vendace (Coregonus albula), three closely related coregonine salmonid fishes. One pair of NOR-bearing chromosomes was found in the peled karyotype. Multichromosomal, but stable, locations of rDNA sites on three pairs of chromosomes were observed in the European whitefish karyotype. Multichromosomal polymorphic locations, both in site and number, were observed in the karyotype of the vendace. Several Ag-, CMA3 and ISH-positive regions were found which defined up to seven cytotypes of five NOR-bearing chromosomes. All positive Ag-NORs detected corresponded both to rDNA ISH- and CMA3-positive signals, which suggests extensive structural polymorphism in the locations of rDNA sites. Stable NOR sites were found at the same location on both homologous elements of the chromosome no. 9 in all individuals, while the remaining NORs were quite variable between individuals, and often present in heterozygous condition. The apparently similar and parallel evolutionary rDNA differentiation patterns in the subfamilies Coregoninae and Salmoninae (family Salmonidae) are observed and discussed. PMID- 11903563 TI - Reproductive processes in two oak (Quercus) contact zones with different levels of hybridization. AB - Patterns of reproductive isolation between two sympatric species of oaks, Quercus gambelii and Q. grisea, that exhibit strong ecological differentiation were examined. A full diallel cross using four trees of each species (i.e. all possible pollinations among eight trees) was performed. This design was repeated at two sites that represent different outcomes of sympatry: (1) a xeric mountain ridge where many hybrids are established (HZ); and (2) a mesic valley bottom where virtually no hybrids are established (MOCYN). By measuring fruit survival at several developmental stages, both the timing and strength of reproductive barriers within and between sites, species, cross types, and pollen dosage levels were examined. In three of four cases, heterospecific fruit set was significantly reduced compared to conspecific fruit set. This reduction occurred after the time of fertilization, but before the onset of embryo growth. Increasing the dose of pollen from an average of 9-194 grains/stigma did not affect this result. Thus, early postfertilization processes play a strong role in species fidelity in these oaks. Quercus gambelii experienced a five-fold decrease in conspecific fruit set at HZ relative to MOCYN. In contrast, heterospecific fruit set of Q. gambelii was the same at both sites. Poor Q. gambelii pollen performance is implicated as playing the major role in this result. One Q. gambelii individual at HZ was highly fecund, and had higher heterospecific than conspecific fruit set; slight introgression in this tree was detected uisng RAPD markers. The Environmental Emasculation Hypothesis that posits that environmental stress can increase the probability of hybrid formation by reducing the competitive ability of male gametes of one species is proposed. PMID- 11903564 TI - The molecular phylogeny of the Sparidae (Pisces, Perciformes) based on two satellite DNA families. AB - In this study, the phylogenetic relationships and which the taxonomic status of the species belonging to the Sparidae family (Pisces: Perciformes) are analysed and revised. This study includes species of this family that are distributed by the North-eastern Atlantic and Mediterranean coasts, is based on the analysis of two satellite DNA families. While one satellite DNA, the centromeric EcoRI family, extends to all the species analysed, the other, the subtelomeric DraI family, is restricted to only six of the 16 species studied. Based on phylogenetic use of these two markers, we conclude that the Sparidae family is composed by two major lineages: one comprising the species of the genera Sparus, Diplodus, Lithognathus, Boops, Sarpa and Spondyliosoma, and one species of Pagellus (P. bogaraveo); and the other lineage is comprised of the species of Pagrus and Dentex, and one species of Pagellus (P. erythrinus). This classification is consistent across the two markers used and clearly contradicts previous morphological phylogenies based mainly on dentition. In addition, the current status and the phylogenetic position of some of the species analysed (i.e. species of Pagrus, Dentex and Pagellus) are not supported by our analyses. Finally, we discuss the value of the morphological characters used until now for the classification of this group of fish. PMID- 11903565 TI - Estimation of linkage disequilibrium for loci with multiple alleles: basic approach and an application using data from bighorn sheep. AB - The great expansion of population genetic data using molecular techniques now allows examination of the extent of linkage disequilibrium for many pairs of loci, each locus often with multiple alleles. The expectation-maximization (EM) algorithm for generating maximum likelihood estimates of gametic frequencies from multiallelic genotypic data is described and applied. The EM algorithm is used in desert bighorn sheep where the population size, and consequently the sample size, is often small. We calculated haplotype frequencies for all pairwise combinations of five major histocompatibility loci and three microsatellite loci in 14 populations; the performance of the algorithm is discussed. Disequilibrium values are calculated and tested for statistical significance. High levels of disequilibrium are found between all pairs of major histocompatibility complex (MHC) loci and between MHC and a linked microsatellite locus. PMID- 11903570 TI - Ways of escape: are all tumours angiogenic? PMID- 11903571 TI - Mucinous (colloid) adenocarcinomas secrete distinct O-acylated forms of sialomucins: a histochemical study of gastric, colorectal and breast adenocarcinomas. AB - AIMS: Mucinous (colloid) adenocarcinomas represent a distinct group of tumours defined by the presence of large amounts of extracellular mucins. By using histochemical methods, we analysed mucins secreted by mucinous versus non mucinous adenocarcinomas and looked for differential secretion profiles. METHODS AND RESULTS: Sixty-four adenocarcinomas were studied (23 colorectal, 17 gastric, and 24 breast tumours). Thirty-two tumours were of the colloid type. The following methods were applied to paraffin tissue sections: (i) Alcian blue (pH 2.5) and periodic acid-Schiff (PAS); (ii) high iron diamine and Alcian blue (pH 2.5); (iii) periodic acid borohydride, potassium hydroxide, and PAS; (iv) periodic acid-thionine Schiff, potassium hydroxide, and PAS; and (v) periodic acid-borohydride and PAS. Most adenocarcinomas secreted acidic mucins, with sialomucins predominating over sulfomucins, except for non-mucinous adenocarcinomas of the breast which showed predominant neutral mucins. All mucinous adenocarcinomas contained C9-O-acyl sialic acid as mono, di(C8,C9)-, or tri(C7,C8,C9)-O-acyl forms. Acidic mucins secreted by the majority of non-colloid adenocarcinomas consisted of non-O-acylated sialomucins. CONCLUSIONS: C9-O acylation of sialic acid is a characteristic feature of mucinous adenocarcinomas and can be readily detected by histochemical methods. PMID- 11903572 TI - Extranodal mantle cell lymphoma mimicking marginal zone cell lymphoma. AB - AIM: We report a case of mantle cell lymphoma masquerading as a marginal zone cell lymphoma. METHODS AND RESULTS: In the initial manifestation in the palatine tonsils, the neoplastic cells were found to grow exclusively within the marginal zones of secondary follicles which showed a preserved mantle zone. The few immunostains performed showed a B-cell phenotype including an immunoglobulin light chain restriction. The extranodal manifestation, the growth pattern, and the immunophenotype led to the diagnosis of an extranodal marginal zone B-cell non-Hodgkin's lymphoma (NHL). The specimen from the relapse occurring 8 months later exhibited diffuse monomorphous cells co-expressing B-cell antigens and CD5, CD43 and cyclin D1, leading to the diagnosis of mantle cell lymphoma. Re investigation of the initial biopsy revealed that the neoplastic cells within the marginal zones had a mantle cell lymphoma immunophenotype expressing cyclin D1, the immunoglobulin heavy chains IgD and IgM and partly CD5. Both lesions harboured identical clonal immunoglobulin gene rearrangements proving that they represented different manifestations of the same lymphoma. CONCLUSION: This case emphasizes the importance of broad immunohistological investigation of B-cell NHLs involving the marginal zone. PMID- 11903573 TI - HSP70 is selectively over-expressed in the blast cells of the germinal centres and paracortex in reactive lymph nodes. AB - AIMS: We evaluated by immunohistochemistry HSP70 expression in reactive lymph nodes since its morphological expression and location have not been previously described and correlated with lymphocyte kinetics. METHODS AND RESULTS: Ninety six cases of non-specific lymphadenitis were immunostained for HSP70, CD20, CD3, Ki67, Bcl-2, CD21. The type and the location of HSP70-positive cells were determined. Their number out of 2000 cells in each germinal centre and in each paracortical area was counted at 60x magnification with the help of a quantitative grid. Seventeen percent of germinal centre cells and 7.6% of the paracortex cells were positive. This difference was highly significant. The positively reacting cells were B-cells and had a blast (centroblast or immunoblast) morphology, with negative mantle and marginal lymphocytes and T cells. Lymphoplasmacytoid cells and plasma cells reacted only weakly or were negative. Germinal centre antigen-presenting cells and interdigitating dendritic cells reacted from lightly to moderately. CONCLUSIONS: HSP70 was selectively over expressed by B-blasts mainly located within germinal centres with a lower number in the paracortex. The difference in the mean number between the two sites was statistically highly significant. No correlation was found with bcl-2 and Ki67 expression. Mantle, marginal and T-lymphocytes were always negative. The biological meaning and role of this over-expression in centroblasts and immunoblasts remain to be elucidated. PMID- 11903574 TI - CD9 immunohistochemical staining of breast carcinoma: unlikely to provide useful prognostic information for routine use. AB - AIMS: CD9, a cell membrane glycoprotein, is found in a variety of tumour cells and is believed to regulate cell motility and possibly cell growth. It has been reported that the absence of CD9 is associated with increased aggressiveness of breast carcinoma, but no detailed studies of the distribution of CD9 in normal and abnormal breast tissue are available. This investigation was aimed at studying the distribution of CD9 in a wide variety of breast biopsies including normal, benign, and malignant cases, and assessing its usefulness as a prognostic marker in breast cancer. METHODS AND RESULTS: Sections of 113 breast biopsies from female and male patients including 10 normal, 23 benign, and 80 malignant cases were examined. The monoclonal antibody CD9 (Novacastra Ltd, Newcastle-upon Tyne, UK) was used with the avidin-biotin complex immunoperoxidase technique. The results were assessed semiquantitatively using a four scale system of 3+, 2+, 1+ and negative. All normal and benign epithelial cells were strongly stained (3+). In female breast carcinomas, 40% were 3+, 49% were 2+, and 11% were 1+. Both cases of male breast carcinomas scored 3+. For female breast cancers, the results were then correlated to tumour grade, the presence or absence of lymph node metastases, and oestrogen and progesterone receptor status. No significant statistical correlation was found with any of these parameters. We then examined 11 axillary lymph nodes with metastases from some of the above cases. Three of these cases had a CD9 score of 3+, seven were 2+, and one was 1+. The metastatic tumours in all 11 cases were strongly stained (3+). CONCLUSIONS: Immunostaining for CD9 is unlikely to provide any useful additional prognostic information for clinical purposes. PMID- 11903575 TI - CD99/MIC-2 surface protein expression in breast carcinomas. AB - AIM: To evaluate the expression of CD99/MIC-2 surface protein in invasive breast carcinomas and demonstrate whether or not there is a relationship with tumour phenotype. METHODS AND RESULTS: Thirty-five invasive breast carcinomas, including five metaplastic carcinomas, were stained with CD99 primary antibodies using standard protocols based on streptavidin-biotin-peroxidase method. Four out of five metaplastic carcinomas expressed CD99/MIC-2 protein, three of them were matrix-producing carcinomas. From the other 30 cases, only an invasive apocrine carcinoma was positive. There was no statistical correlation between CD99 expression and the parameters analysed (histological typing and grading, proliferative index and nodal status). CONCLUSIONS: CD99/MIC-2 is expressed in breast carcinomas, especially in the matrix-producing variant of metaplastic carcinomas, which impairs its use as a marker to differentiate metaplastic carcinomas from primary and metastatic sarcomas of the breast. It seems to have no prognostic implications. However, phenotype similarities with other chondromyxoid tumours that also express the protein, like mesenchymal chondrosarcomas, suggest a relationship between MIC-2 reactivity and morphological differentiation. PMID- 11903576 TI - Malignant epithelioid mesothelioma: anti-mesothelial marker expression correlates with histological pattern. AB - AIMS: Malignant epithelioid mesothelioma shows marked cytoarchitectural diversity. The aim of the study was to evaluate how immunoreactivity with mesothelial markers related to histological pattern. METHODS AND RESULTS: Ninety two cases of malignant epithelioid mesothelioma (60 pleural, 32 peritoneal) were examined and classified as exhibiting tubulopapillary, adenomatoid, solid, small cell or pleomorphic patterns. All cases were immunohistochemically stained with thrombomodulin, calretinin, CD44H, and cytokeratin 5/6. Cases of malignant mesothelioma exhibited a number of different histological forms. Immunohistochemical expression of each mesothelial marker tested was not homogeneous across different histological patterns of malignant epithelioid mesothelioma, even within the same tumour section. Calretinin (with nuclear expression) was identified to show the highest overall sensitivity and lowest range variation in staining (67% sensitivity in small cell areas to 100% expression in pleomorphic areas). Cytokeratin 5/6 and thrombomodulin yielded similar overall sensitivity. Thrombomodulin appeared to demonstrate higher sensitivity for small cell variant tumour (83% sensitivity). A notable advantage with cytokeratin 5/6 was that expression was more diffuse in nature rather than the focal membranous elaboration seen in thrombomodulin. The widest range of staining was seen in small cell mesothelioma (83% sensitivity with thrombomodulin to 17% sensitivity with cytokeratin 5/6) and in tubulopapillary areas (90% sensitivity with calretinin to 38% sensitivity with CD44H). CONCLUSIONS: Calretinin appears most useful and shows the highest overall sensitivity for malignant epithelioid mesothelioma, with good expression in areas displaying a tubulopapillary, adenomatoid, solid and pleomorphic pattern. For small cell mesothelioma, thrombomodulin appears to confer higher sensitivity and is advocated, in this setting, as the first line mesothelial marker. Cytokeratin 5/6 is a useful and easily interpretable mesothelial marker. CD44H is not of particular use in the diagnosis of malignant epithelioid mesothelioma. Accurate interpretation of immunohistochemistry in mesothelioma requires an awareness of the immunophenotypic heterogeneity identified in different histological forms of the tumour, and this is of particular importance in small biopsies. PMID- 11903577 TI - Expression of cell cycle regulatory proteins in the multistep process of oesophageal carcinogenesis: stepwise over-expression of cyclin E and p53, reduction of p21(WAF1/CIP1) and dysregulation of cyclin D1 and p27(KIP1). AB - AIMS: Cell cycle regulatory proteins were analysed by immunohistochemistry in order to clarify how their expression changes with the degree of atypia as oesophageal surface squamous epithelium progresses from normal mucosa, through reactive change, low-grade dysplasia, and high-grade dysplasia to mucosal invasive carcinoma. METHODS AND RESULTS: Immunostaining for cyclin D1, cyclin E, p21, p27, p53 and Ki67 proteins was performed using 22 normal mucosa, 17 reactive change, 22 low-grade dysplasia, 15 high-grade dysplasia and 22 mucosal invasive carcinoma specimens. Normal mucosa, low-grade dysplasia and high-grade dysplasia samples were taken from patients without any oesophageal invasive carcinoma by endoscopic biopsy or endoscopic mucosal resection, and reactive change and mucosal invasive carcinoma were obtained from oesophagectomy material. Stepwise over-expression of cyclin E (P < 0.0001) and p53 (P < 0.0001), reduction of p21 (P=0.0189) and dysregulation of cyclin D1 and p27 were observed in the multistep process of oesophageal carcinogenesis. Significant differences in expression of p27 (P < 0.0001), p53 (P=0.0299) and Ki67 (P=0.0101) were observed between reactive change and low-grade dysplasia. Furthermore, expression of cyclin D1, cyclin E, p27 and p53 in mucosal invasive carcinoma were significantly different from those in high-grade dysplasia (P=0.0079, P=0.0237, P=0.0042 and P= 0.0299, respectively). CONCLUSIONS: Cell cycle regulatory proteins, cyclin E, p53 and p21 show stepwise over-expression or reduction with progression of oesophageal carcinogenesis, correlating with the increased cell proliferation observed with Ki67 labelling. We conclude that immunohistochemical analysis for p27, p53 and Ki67 is practically useful for the discrimination between low-grade dysplasia and reactive change. Cyclin D1, cyclin E, p27 and p53 help to distinguish high-grade dysplasia from mucosal invasive carcinoma. PMID- 11903578 TI - Expression of MMP-2 is associated with progression and lymph node metastasis of gastric carcinoma. AB - AIMS: One important step in tumour invasion is the penetration of the basement membrane. Matrix metalloproteinases (MMPs) play a key role in the migration of normal and malignant cells through the basement membrane. The aim of this study was to investigate correlations between matrix metalloproteinase 2 (MMP-2) immunoreactivity and currently used classification systems and possible relationships between lymph node metastasis and MMP-2 expression. METHODS AND RESULTS: This prospective study analysed specimens obtained from 114 gastric cancer patients (mean age 64 years; range 33-86 years) who underwent gastrectomy with extended lymphadenectomy. All specimens were categorized according to UICC classification, WHO classification, tumour differentiation, Lauren classification, Ming classification and Goseki classification. Formalin-fixed paraffin-embedded tumour specimens were stained using an avidin-biotin complex peroxidase assay. MMP-2 expression in the tumour epithelium was studied by immunohistochemistry with semiquantitative (score 0-3) evaluation. The MMP-2 staining pattern was positive (score 1-3) in 93 (81.6%) specimens and negative (score 0) in 21 (18.4%) samples. No significant correlations were found between MMP-2 expression and other variables such as age, tumour differentiation, WHO, Lauren, Goseki, and Ming classifications. In contrast, the intensity of MMP-2 staining in tumour cells correlated significantly with depth of tumour infiltration (T-stage), lymph node metastasis (N-stage), distant metastasis (M stage), and UICC stage. CONCLUSIONS: Expression of MMP-2 is strongly associated with tumour progression and lymph node metastasis in gastric cancer. Therefore MMP-2 staining may be clinically useful as predictor of tumour progression, especially for lymph node metastasis. PMID- 11903580 TI - Adenomatous polyposis coli (APC), beta-catenin, and cadherin are expressed in human bone and cartilage. AB - AIMS: Members of the cadherin and catenin families are involved in chondrogenesis and catenin gene mutations have been detected in malignant tumours of bone. This study was undertaken to assess in detail expression of cadherin, beta-catenin and the associated tumour suppressor gene product APC in bone and cartilage at different stages of human skeletal maturity and in non-neoplastic and neoplastic osteoarticular disease. METHODS AND RESULTS: Immunohistochemical staining of formalin-fixed paraffin-embedded normal and osteoarthritic adult articular cartilage, fetal growth plate and a series of tumours of bone and cartilage was undertaken with a panel of antibodies against APC, beta-catenin, and pan cadherin. This study demonstrated expression of APC, beta-catenin and cadherin in normal and diseased bone and cartilage. APC was present both in osteoblasts and osteoclasts but not in osteocytes. Although only weak APC staining of occasional growth plate hypertrophic chondrocytes and normal articular chondrocytes was seen, APC staining was increased in osteoarthritic articular cartilage. beta catenin and pan-cadherin staining was strongly positive in osteoclasts and osteoblasts, with expression being lost when bone cells differentiated into osteocytes. Expression of APC, beta-catenin and pan-cadherin in bone tumours was similar to that of non-neoplastic adult tissues. CONCLUSIONS: These findings suggest previously unrecognized roles for APC in regulation of function of chondrocytes, osteoblasts and osteoclasts and support the view that catenin cadherin interactions are important in regulation of bone cell activity. Abnormalities of expression or function of these molecules may be important in formation of bone tumours and their clinical behaviour. PMID- 11903581 TI - Arthroplasty-associated malignant fibrous histiocytoma: two case reports. AB - AIMS: Sarcoma localized to the site of an arthroplasty procedure is a rare occurrence, and detailed histological depictions and descriptions are limited. We report the clinicopathological findings in two cases of arthroplasty-associated malignant fibrous histiocytoma (MFH) and review the literature. METHODS AND RESULTS: The patients were an elderly man and woman. Medical histories, radiographs and slides were reviewed. Immunohistochemistry, electron microscopy, cytogenetics, and electron dispersion spectroscopy were performed in one case. Both were destructive femoral bone tumours that appeared 2 and 8 years post-total hip arthroplasty, and pursued aggressive clinical courses. The histology was similar in both tumours, consisting of high-grade, pleomorphic sarcoma with numerous osteoclastic giant cells, prominent phagocytic activity, and entrapped particles of bone cement. Literature review disclosed 14 previous reports of arthroplasty-associated MFH, representing the most common phenotype. A number of materials and factors related to arthroplasty procedure, such as metal corrosion, wear debris, osteonecrosis, and chronic inflammation, have been implicated as causative agents. CONCLUSIONS: Arthroplasty-associated MFH is a rare and aggressive tumour. Although the aetiology remains unclear, the small number of arthroplasty-associated sarcomas compared with the large number of joint replacement operations performed over the past four decades suggests a coincidental as opposed to a causal relation. PMID- 11903583 TI - Panel for distinction between mesothelioma and adenocarcinoma: confidence limits for a small study. PMID- 11903579 TI - Combined examination of p27(Kip1), p21(Waf1/Cip1) and p53 expression allows precise estimation of prognosis in patients with gastric carcinoma. AB - AIMS: In order to estimate the prognostic values of p27(Kip1), p21(Waf1/Cip1), and p53, alone and in combination, we investigated immunohistochemically the expression of p27(Kip1), p21(Waf1/Cip1), and p53 proteins in gastric carcinomas. METHODS AND RESULTS: The expression of p27(Kip1), p21(Waf1/Cip1), and p53 was immunohistochemically examined in 140 gastric carcinomas. Positive expression of p27(Kip1) and p21(Waf1/Cip1) correlated significantly with a favourable prognosis (P < 0.05), whereas, positive expression of p53 tended to correlate with poor prognosis. Multivariate survival analysis revealed that TNM stage of tumour (P < 0.001), lymph node state (P=0.005), and p27(Kip1) expression (P=0.006) were independent prognostic factors. A striking stratification of mortality rate was found when patients were divided into four groups according to the expression of p21(Waf1/Cip1) and p27(Kip1). The mortality rate was higher in patients with both p21(Waf1/Cip1)- and p27(Kip1)-negative gastric carcinoma than in patients with one or both positive carcinomas (P < 0.01). In addition, if the four p21(Waf1/Cip1)/p27(Kip1) groups were compared based on p53 status, p53+ cases tended to have a higher mortality rate than p53- cases. CONCLUSION: Our results suggest that low expression of both p27(Kip1) and p21(Waf1/Cip1), could be useful as markers of poorer prognosis, and the combined examination of p27(Kip1), p21(Waf1/Cip1) and p53 expression allows reliable estimation of prognosis for patients with gastric carcinoma. PMID- 11903582 TI - Secondary malignant giant-cell tumour of bone: molecular abnormalities of p53 and H-ras gene correlated with malignant transformation. AB - AIMS: We report two cases of secondary malignant giant-cell tumour occurring without irradiation therapy. To elucidate the mechanism of malignant transformation in this tumour, we searched for the molecular abnormalities of p53, MDM2 and the H-ras genes. METHODS AND RESULTS: These cases were retrieved after a review of 103 cases of primary giant-cell tumour of bone, registered in our institute. One case occurred in the distal femur of a 42-year-old female after surgical curettage, while the other arose in the acetabulum of a 25-year old male after en bloc resection. Microscopically, the malignant tumour in the distal femur was composed of a proliferation of ovoid or fusiform cells arranged in fascicles with high mitotic activities. The malignant transformed tumour in the acetabulum was made up of pleomorphic tumour cells with atypical mitoses. In the tumour of the distal femur, both p53 and H-ras mutations were detected. Abnormal nuclear accumulation of p53 protein and c-myc expression were also revealed by immunohistochemistry. In both cases, the recurrent malignant tumour over-expressed MMP-9 and revealed a higher MIB-1-labelling index compared with the primary conventional giant-cell tumour. CONCLUSIONS: Our results suggest that multiple oncogene or tumour suppressor gene mutations may play an important role during malignant transformation in conventional giant-cell tumours. PMID- 11903585 TI - Retrorectal pseudotumour induced by Ivalon. PMID- 11903586 TI - Proximal epithelioid sarcoma--a misnomer. PMID- 11903587 TI - Iron-induced oesophageal injury and pigmentation. PMID- 11903588 TI - Endosalpingiosis of the colon and appendix. PMID- 11903589 TI - A bright yellow stromal nodule within a uterine leiomyoma: a case report. PMID- 11903590 TI - Naevus cell aggregates in numerous lymph nodes in a case of mucinous mammary carcinoma. PMID- 11903593 TI - Ki67 protein: the immaculate deception? AB - This article updates our previous review of Ki67 published in Histopathology 10 years ago. In this period the numbers of papers published featuring this antibody has increased 10-fold from 338 to 3489 indicating the considerable enthusiasm with which this antibody has been studied. This review attempts to provide an update on the characterization of the Ki67 protein, its function and its use as a prognostic or diagnostic tool. PMID- 11903595 TI - Splenic small B-cell lymphoma with predominant red pulp involvement: a diffuse variant of splenic marginal zone lymphoma? AB - AIMS: Splenic marginal zone lymphoma (SMZL) has been characterized by a micronodular pattern of infiltration, biphasic cytology, follicular replacement and the presence of marginal zone differentiation. Here we describe four cases with some distinctive features, such as diffuse splenic infiltration, lack of micronodules, marginal zone cytology, p53 inactivation and cutaneous involvement. METHODS AND RESULTS: In the course of a review of cases of SMZL, we recognized the existence of a subset of four cases of splenic B-cell lymphoma, with predominantly red pulp involvement, absence of follicular replacement, and a monomorphous population of tumoral cells resembling marginal zone B-cells, with scattered nucleolated blast cells. The immunophenotype (bcl2+, CD5-, CD10-, CD43 , CD23-, cyclin D1-, IgD- (3/4)) was consistent with SMZL. Bone marrow infiltration (4/4) and peripheral blood involvement (2/4) showed similar findings to those described for SMZL in these locations. However, unlike classical SMZL, 2/4 had cutaneous involvement, and 4/4 cases showed either p53 mutation or anomalous p53 staining (p53+, p21-). CONCLUSIONS; In spite of a diffuse pattern of splenic infiltration, cutaneous involvement and p53 alterations, these cases have findings that overlap with those corresponding to classic SMZL (symptomatology, morphology of bone marrow, lymph nodes, peripheral blood involvement, and immunophenotype). We suggest that these cases be considered a putative variant of SMZL. PMID- 11903594 TI - Accessory cell tumour: a clinicopathological study of 16 aggressive tumours containing EBV-positive Hodgkin and Reed-Sternberg-like giant cells. AB - AIMS: Lymph nodes contain non-lymphoid accessory cells including follicular dendritic cells and interdigitating dendritic cells. Functionally, these cells belong to the category of immune accessory cells involved in antigen presentation to B or T-lymphocytes. Neoplastic proliferation of these cells is very uncommon. We present here the clinicopathological features of 16 cases of accessory cell tumour. METHODS AND RESULTS: We performed electron microscopic and immunohistochemical examinations, and used in-situ hybridization for EBV-encoded RNA (ISH-EBV) to detect the EBV genome in 11 cases, and Southern blot analysis to assess EBV clonality in two cases. Tumour cells were composed of oval-to-spindle cells arranged in diffuse, vague storiform, fascicular and sometimes whorled patterns in a background of small lymphocytes. In all cases, binucleated or multinucleated Hodgkin and Reed-Sternberg-like giant cells were encountered. Staining for CD68 was positive in all cases. CD21, CD35, Ki-M4p, Ki-FDC1p, and S100 exhibited variable reactivity. ISH-EBV yielded positive labelling in seven of 11 cases, of which five exhibited EBV only in Hodgkin and Reed-Sternberg-like giant cells. Southern blot analysis showed clonality of EBV terminal repeats (EBV TR) in the two cases examined. Electron microscopic examination showed that many of the tumour cells had numerous interwoven long villous cell processes connected by occasional desmosomes. Many tumours were very refractory to chemotherapy and radiation, with a few exceptions, and half of the cases classified initially as stage IV. A short survival time, of 10 months or less, was observed in seven of 16 patients. CONCLUSIONS: Our study identified more aggressive behaviour of accessory cell tumours. Our results suggest that EBV may potentially induce activation of accessory cells to form Hodgkin and Reed-Sternberg-like giant cells, which correspond with poor prognosis. PMID- 11903596 TI - Histiocyte-rich, T-cell-rich B-cell lymphoma: a distinct diffuse large B-cell lymphoma subtype showing characteristic morphologic and immunophenotypic features. AB - AIMS: The clinicopathological features of histiocyte-rich, T-cell-rich B-cell lymphoma (HRTR-BCL) were first recognized in 1992. In this study, 60 cases of HRTR-BCL were analysed in order to provide a detailed morphological and immunophenotypical profile of the disorder. METHODS AND RESULTS: HRTR-BCL is easily distinguished from other B-cell lymphomas rich in stromal T-cells by (i) a diffuse or vaguely nodular growth pattern, (ii) the presence of a minority population of CD15-, CD20+ large neoplastic B-cells, (iii) a prominent stromal component composed of both T-cells and non-epithelioid histiocytes, and (iv) the scarcity of small reactive B-cells. These criteria also enable a reliable distinction from lymphocyte-rich classical Hodgkin's lymphoma (CHL), from lymphocyte-predominant Hodgkin's lymphoma (LPHL), paragranuloma type and from peripheral T-cell lymphoma. Based on the morphology of the neoplastic cells and on their frequent bcl-6 immunoreactivity, we speculate that HRTR-BCL may be derived from a progenitor cell of germinal centre origin. CONCLUSIONS: HRTR-BCL presents characteristic clinical features, affecting predominantly middle-aged men who present with advanced stage disease and are at high risk of treatment failure. Considering these distinctive clinicopathological features, recognizing HRTR-BCL as a lymphoma entity may be justified. PMID- 11903597 TI - Thymic sarcomatoid carcinoma with skeletal muscle differentiation: report of two cases, one with cytogenetic analysis. AB - AIMS: Malignant thymic tumour histologically resembling a soft tissue sarcoma is extremely rare and defined as sarcomatoid carcinoma in the recent World Health Organization (WHO) classification. We report two such cases in which the tumour cells showed a prominent rhabdomyoblastic differentiation and analyse whether these tumours retain an epithelial nature at least in part. METHODS AND RESULTS: One tumour occurred in a 51-year-old man (Case 1) and the other in a 40-year-old woman (Case 2). Microscopically, both tumours consisted essentially of two types of tumour cells: spindle and large round cells, with no apparent epithelial components. Osteosarcomatous small foci were also found in Case 2. Immunohistochemically, desmin and muscle-specific actin were positive in the majority of both types of tumour cells, whereas myogenin was predominant in the spindle cells and myoglobin in the large round cells. Some of both types of cells expressed cytokeratin with co-expression of myoglobin in the large round cells, but with no myogenin in the spindle cells. Some cytokeratin-positive spindle cells were also negative for desmin. Ultrastructural examination of a recurrent tumour in Case 2 revealed some epithelial features among the spindle cells. Cytogenetic study of the same tumour showed a complex abnormality including der(16)t(1;16)(q12;q12.1), an identical pattern previously reported in a case of thymic squamous cell carcinoma. CONCLUSIONS: The findings support the definition in the WHO classification of sarcomatoid carcinoma that includes purely sarcomatous tumour as in the present cases. Occurrence of this type of tumour may indicate a relationship between thymic epithelial cells and myoid cells and/or a potential for divergent differentiation in thymic epithelial tumours. PMID- 11903598 TI - Sarcomatoid/metaplastic carcinoma of the breast: a clinicopathological study of 12 cases. AB - AIMS: To analyse the clinical and pathological features with long-term follow-up of a series of 12 cases of sarcomatoid carcinoma of the breast. methods and results: The cases were selected from the surgical files of the Department of Pathology, University of Edinburgh, between 1977 and 1988. The following clinical parameters were recorded: the age of the patients, size of tumour, presence or absence of lymph node or distant metastases, and patient survival. Pathological assessment included: the type of epithelial and mesenchymal components, the proportion of monophasic to biphasic tumours and the presence of adjacent in-situ carcinoma/atypical epithelial proliferation. The mean age of the patients was 61 years with a median of 64 and range 46-82 years. The mean size of the tumour was 52 mm (range 22-100 mm). None of the patients had distant metastasis at presentation and only one case had local lymph node metastasis which had a carcinomatous appearance. Five women were still alive after a minimum 12-year follow-up period. Four patients died of their disease (three with lung metastasis only and one with lung and bone metastases), one died of carcinoma of the cervix and two patients were lost to follow-up. Pathologically, four cases (33.3%) had no or almost undetectable epithelial structures by light microscopy, i.e. "monophasic sarcomatoid carcinoma". The remaining cases revealed varying proportions of both epithelial and mesenchymal elements, i.e. "biphasic sarcomatoid carcinoma". Of the epithelial component, six (50%) tumours had predominantly carcinoma of no special type, one lobular and one tubular carcinoma. The mesenchymal component was fibromatosis/nodular fasciitis-like, malignant fibrous histiocytoma-like (MFH), osteosarcoma-like and fibrosarcoma like in five (42%), four (33%), two (17%) and one (8%) tumours, respectively. In 3/4 monophasic tumours, the mesenchymal component was of a low-grade fibromatosis/nodular fasciitis type. In 6/12 (50%) of the cases there was associated in-situ atypical epithelial proliferation (five ductal carcinoma in situ (DCIS) and one atypical ductal hyperplasia). CONCLUSIONS: From this small series it appears that sarcomatoid carcinoma is an uncommon tumour, which is large in size and tends to lack local or distant metastasis at presentation. Pathologists should be alert to the presence of the bland monophasic sarcomatoid carcinoma which has a pure mesenchymal appearance on light microscopy, but epithelial components demonstrated by cytokeratin immunohistochemistry. These showed metastases on long-term follow-up, similar to other histological patterns of sarcomatoid carcinoma. PMID- 11903599 TI - Malignant mesenchymoma of the uterus, arising in a leiomyoma. AB - AIMS: To document and find evidence for the rare occurrence of malignant progression of a benign uterine leiomyoma with divergent mesenchymal differentiation. In a 54-year-old female a large pedunculated tumour was encountered which had suddenly increased in size and had apparently arisen at the site of a subserosal uterine leiomyoma first described 19 years earlier. The tumour seemed histologically diverse in its composition. The possibility of malignant progression of the benign leiomyoma with divergent mesenchymal differentiation was entertained. METHODS AND RESULTS: In the resected tumour, smooth muscle, osseous and adipose components were identified and these were assessed using recognized histological criteria of malignancy. In addition to a benign smooth muscle component, malignant leiomyosarcomatous tissue was seen in addition to a malignant osseous component and a malignant adipose component. The tumour thus met the criteria of a malignant mesenchymoma. The relationship of the different components of the tumour was analysed by immunohistochemistry and with molecular loss of heterozygosity (LOH) analysis. In the osseous and leiomyosarcomatous components a similar LOH pattern was observed. The adipose component showed a distinct LOH pattern. Retention of smooth muscle differentiation in the osseous component was demonstrated by desmin immunostaining. CONCLUSION: Malignant transformation of benign uterine leiomyoma may rarely occur. Mesenchymal stem cells underlying these tumours may show divergent mesenchymal differentiation. PMID- 11903600 TI - Fatty acid synthase is highly expressed in carcinoma, adenoma and in regenerative epithelium and intestinal metaplasia of the stomach. AB - AIMS: To investigate the relation of fatty acid synthase (FAS) expression to the clinicopathological characteristics of gastric cancers and gastric tumorigenesis. METHODS AND RESULTS: FAS expression was examined immunohistochemically in 626 gastric cancers, 51 gastric adenomas, and non-neoplastic epithelium contiguous with cancer tissue including normal foveolae, intestinal metaplasia, regenerative epithelium, and gastric glands. FAS expression was found in more than 70% of gastric cancers. Interestingly, it occurred preferentially in differentiated carcinomas, male cases, and in patients over 51 years old. Although previous reports showed that FAS expression is closely related to cancer progression, in gastric cancers FAS expression had no relationship with prognosis, cancer progression as indicated by depth of invasion, venous and lymphatic permeation, and distant metastasis. Gastric tubular adenoma and intestinal metaplasia, which are thought to be associated with well-differentiated gastric carcinomas, showed a frequency of FAS expression similar to that of differentiated carcinomas; however, normal foveolae and gastric glands showed no or less FAS expression. CONCLUSIONS: FAS expression occurs at the early stage of tumorigenesis and plays important roles in the formation of precancerous foci rather than in the progression of carcinoma of the stomach. PMID- 11903602 TI - Multifocal distribution of pulmonary capillary haemangiomatosis. AB - AIMS: We investigated a case of pulmonary capillary haemangiomatosis, a rare condition, to determine the extent of the pathological changes within the lungs. Systematic histological sampling has not previously been performed in this condition. METHODS AND RESULTS: A 52-year-old woman with a history of ischaemic cardiomyopathy suffered from repeated respiratory infections, which were attributed to chronic pulmonary congestion. She died suddenly of fulminant pulmonary thromboembolism. An autopsy was performed and lung tissue was sampled at multiple sites. Beside passive congestion, the lungs showed well-circumscribed areas containing proliferations of small capillaries infiltrating the pulmonary septa and the walls of otherwise normal blood vessels and bronchi. The most severely affected areas were found to be in the periphery of both lower lobes. A diagnosis of pulmonary capillary haemangiomatosis was made. CONCLUSIONS: This is the first case of pulmonary capillary haemangiomatosis in which systematic histological sampling has been performed. Mapping of lesions disclosed the multifocal distribution of pulmonary capillary haemangiomatosis in this patient. PMID- 11903601 TI - Galectin-3, a marker of well-differentiated thyroid carcinoma, is expressed in thyroid nodules with cytological atypia. AB - AIMS: The distribution of galectin-3, a widely recognized marker of well differentiated thyroid carcinoma, was investigated in 95 thyroid lesions including nodules with foci of cytoarchitectural atypia. METHODS AND RESULTS: Twenty-eight papillary carcinomas, five follicular carcinomas, one Hurthle cell carcinoma, three poorly differentiated carcinomas, one anaplastic carcinoma, 25 nodular hyperplasias and 27 follicular adenomas, including nodules with atypical features, three neoplasms of undetermined malignant potential and two thyroiditis cases were examined. By immunohistochemistry, galectin-3 was consistently found in carcinomas; otherwise benign nodules exhibited galectin-3-positive clusters of cells with poorly developed features of differentiated carcinoma (mainly of papillary type) such as nuclear chromatin clearing, nuclear clefting, pseudoinclusions, which, in each case, were not histologically sufficient to warrant a definitive diagnosis of malignancy. In other nodules galectin-3 staining was negative. The latter were either clearly benign or showed constantly a minor degree of chromatin clearing and of other atypical features when compared with galectin-3-positive cases. CONCLUSIONS: Galectin-3, a reliable marker of differentiated thyroid carcinoma as confirmed in our series of malignant neoplasms, appears expressed in nodules with an overall benign appearance but with focal areas suspicious for malignancy. The significance of such findings needs to be further investigated. PMID- 11903603 TI - Patterns of episialin/MUC1 expression in endometrial carcinomas and prognostic relevance. AB - AIMS: To investigate episialin/MUC1 expression in the normal, hyperplastic and neoplastic endometrium, and relate patterns of tumour MUC1 reactivity with histopathological characteristics, oestrogen and progesterone receptor (ER and PR) status, bcl-2 and p53 oncoproteins and with clinical behaviour. METHODS AND RESULTS: We studied 42 normally cycling endometria, 45 endometrial hyperplasias of various forms, and 111 endometrial carcinomas of endometrioid and non endometrioid cell types with specific monoclonal antibodies employing standard immunohistochemical techniques. The follow-up period ranged from 34 to 182 months with a median of 86 months. Epithelial mucin episialin/MUC1 was consistently expressed in the normal endometrium, following a cyclical pattern: "apical membrane staining" in early and mid-proliferative endometrium; "purely cytoplasmic staining" in late proliferative endometrium; and "cytoplasmic staining with intraluminal secretions" in secretory endometrium. Immunostaining patterns in simple and complex hyperplasia were similar to late proliferative endometrium, while atypical hyperplasias and endometrial carcinomas either simulated patterns of proliferative endometrium or lacked MUC1 reactivity. Membranous MUC1 positivity was statistically more frequent in endometrioid carcinomas compared with carcinomas of non-endometrioid type (P = 0.006). Cytoplasmic MUC1 positivity was significantly associated with poor prognosis, while MUC1-negative carcinomas were associated with PR expression and an improved survival (P=0.04). There was no association of MUC1 patterns with bcl-2 and p53 immunoreactivity or with other histopathological variables. CONCLUSIONS: Episialin/MUC1 is an integral component of the normal premenopausal endometrium and is probably hormonally regulated. It is frequently expressed in endometrial hyperplasias and carcinomas. The loss of MUC1 expression from endometrial carcinomas is associated with a favourable prognosis. PMID- 11903604 TI - Lesson of the month. An unusual brain "tumour". PMID- 11903605 TI - Angiogenesis in lymph node metastases. PMID- 11903607 TI - Lack of angiogenesis in lymph node metastases of carcinomas is growth pattern dependent. PMID- 11903608 TI - bcl-2 and p21 immunostaining of cervical tubo-endometrial metaplasia. PMID- 11903610 TI - Apoptosis and Rushton body formation. PMID- 11903609 TI - Immunophenotypic and genotypic analysis of a case of primary peripheral primitive neuroectodermal tumour (pPNET) of the urinary bladder. PMID- 11903611 TI - Prostatic lymphoma of mucosa-associated lymphoid tissue: an uncommon location. PMID- 11903612 TI - Immunopathogenesis of hepatitis C virus infection. AB - Hepatitis C virus, a recently identified member of the family Flaviviridae, is an important cause of chronic viral hepatitis and cirrhosis. There are similarities in the nature of the immune response to this pathogen with immunity in other flavivirus and hepatotropic virus infections, such as hepatitis B. However, the high rate of viral persistence after primary hepatitis C infection, and the observation that neutralizing antibodies are not protective, would suggest that there are a number of important differences between hepatitis C, other flaviviruses, and hepatitis B. The phenomenon of quasispecies evolution and other viral factors have been proposed to contribute to immune evasion by hepatitis C virus. In the face of established persistent infection, virus-specific cytotoxic T lymphocytes may exert some control over viral replication. However, these same effectors may also be responsible for the progressive liver damage characteristic of chronic hepatitis C infection. The nature of protective immunity, including the role of innate immune responses early after hepatitis C exposure, remains to be defined. PMID- 11903613 TI - DNA vaccines: future strategies and relevance to intracellular pathogens. AB - Increasing awareness of microbial threat has rekindled interest in the great potential of vaccines for controlling infectious diseases. The fact that diseases caused by intracellular pathogens cannot be overcome by chemotherapy alone has increased our interest in the generation of highly efficacious novel vaccines. Vaccines have proven their efficacy, as the immunoprotection they induce appears to be mediated by long-lived humoral immune responses. However, there are no consistently effective vaccines available against diseases such as tuberculosis and HIV, and other infections caused by intracellular pathogens, which are predominantly controlled by T lymphocytes. This review describes the T-cell populations and the type of immunity that should be activated by successful DNA vaccines against intracellular pathogens. It further discusses the parameters that need to be fulfilled by protective T-cell Ag. We then discuss future approaches for DNA vaccination against diseases in which cell-mediated immune responses are essential for providing protection. PMID- 11903614 TI - Ultraviolet light induced injury: immunological and inflammatory effects. AB - This article reviews many of the complex events that occur after cutaneous ultraviolet (UV) exposure. The inflammatory changes of acute exposure of the skin include erythema (sunburn), the production of inflammatory mediators, alteration of vascular responses and an inflammatory cell infiltrate. Damage to proteins and DNA accumulates within skin cells and characteristic morphological changes occur in keratinocytes and other skin cells. When a cell becomes damaged irreparably by UV exposure, cell death follows via apoptotic mechanisms. Alterations in cutaneous and systemic immunity occur as a result of the UV-induced inflammation and damage, including changes in the production of cytokines by keratinocytes and other skin-associated cells, alteration of adhesion molecule expression and the loss of APC function within the skin. These changes lead to the generation of suppressor T cells, the induction of antigen-specific immunosuppression and a lowering of cell-mediated immunity. These events impair the immune system's capacity to reject highly antigenic skin cancers. This review gives an overview of the acute inflammatory and immunological events associated with cutaneous UV exposure, which are important to consider before dealing with the complex interactions that occur with chronic UV exposure, leading to photocarcinogenesis. PMID- 11903615 TI - Induction of CD8+ T-lymphocyte responses to a secreted antigen of Mycobacterium tuberculosis by an attenuated vaccinia virus. AB - Protective immunity against Mycobacterium tuberculosis infection requires the activation of mycobacterium-specific CD8+ T cells, as well as CD4+ T cells. Therefore, optimizing strategies that stimulate CD8+ T cells recognizing dominant mycobacterial antigens, including secreted proteins, may lead to the development of more effective vaccines against tuberculosis. To generate a viral vaccine that is safe in humans, the early secreted protein, MPT64, was expressed in the attenuated vaccinia virus (VV) strain, modified vaccinia virus Ankara (MVA-64). The immunogenicity of MVA-64 was compared with that of the Western Reserve strain of VV (VVWR-64). The replication-defective MVA-64 was as efficient as VVWR-64 in inducing specific antibodies and cytolytic T-cell responses to a defined H-2-Db restricted epitope on MTP-64. In addition, priming with MPT64-expressing plasmid DNA (DNA-64), and boosting with either MVA-64 or VVWR-64, markedly enhanced MPT64 specific cytolytic and IFN-gamma-producing CD8+ T-cell responses. These findings suggest that MVA may be a suitable vaccine carrier for stimulating mycobacterium specific CD8+ T-cell responses and may be particularly relevant for developing vaccines for use in regions endemic for tuberculosis and HIV infection. PMID- 11903616 TI - A locus affecting immunoglobulin isotype selection (Igis1) maps to the MHC region in C57BL, BALB/c and NOD mice. AB - We have previously shown that H2b mice with B10 or BALB genetic backgrounds have higher basal levels of IgG2a than H2k and H2d congenic strains and, hence, have low IgG1/IgG2a ratios, which is consistent with a T1 cytokine milieu. The phenotypic marker of the high IgG2a levels, denoted immunoglobulin isotype-1 (Igis1) was provisionally mapped telomeric of IEbeta using MHC recombinant mice. In addition, data from B10.A(2R), B10.A(1R) and B10.A(18R) mice indicated that Igis1 may lie in a 27 kb region between G7b (Sm-X5) and G7c. In the present study we confirm that Igis1 is in the H2 region using BALB and B10 congenic F2 mice. H2bb F2 mice had higher IgG2a levels than the H2dd parental strains. H2bd F1 and F2 mice on the B10 background produced IgG2a levels comparable with the H2bb parental strain, indicating that the b allele was dominant. In contrast, the H2bd F1 and F2 mice on the BALB background produced IgG2a levels between those of the H2bb and H2dd parental strains, indicating codominance of the b and d alleles. This suggests that a background gene influences regulation of IgG2a levels by Igis1. Non-obese diabetic (NOD) mice (KdIAg7IEnu11Db), which can develop type 1 diabetes, had higher levels of IgG2a than NOD-H2d congenic mice. Thus, Igis1 affects isotype selection in the presence of non-MHC diabetes genes. As type 1 diabetes is associated with T1 responses, Igis1 may affect susceptibility to this condition. PMID- 11903617 TI - Changes in behaviour, cortisol and lymphocyte types during isolation and group confinement of sheep. AB - This experiment compared changes in complex behaviour patterns, adrenal corticosteroid secretion and the numbers of various types of lymphocytes in sheep that were subjected to the stress of confinement. Grazing Merino ewes (n=80 in five replicated experiments) were confined either in groups of four per pen or in total isolation from other sheep. The percentage of CD4+ lymphocytes increased while the percentage of CD8+ lymphocytes decreased over the experimental period. This result was more pronounced in isolated sheep than in grouped sheep. The increase in CD4:CD8 was greater for isolated sheep than for grouped sheep and greater for 2 week sheep than for 3 week sheep. The percentage of CD5+ cells also increased, less so in isolated than in grouped animals. Interpreting these changes as a recovery of immune competence following introduction of a stressor, it is apparent that isolation impaired immune system recovery more severely than group confinement. Physiological and behavioural adaptation over the period were characterized by a decline in the adrenocortical response, resumption of the normal pattern of flocking behaviour and a reduction in motor activity during the test. These findings add to the evidence pointing to the possible correspondence between critical features of the psychoneural, neuroendocrine and immune systems. PMID- 11903619 TI - B lymphocytes not required for progression from insulitis to diabetes in non obese diabetic mice. AB - Previous studies have implicated B lymphocytes in the pathogenesis of diabetes in the non-obese diabetic (NOD) mouse. While it is clear that B lymphocytes are necessary, it has not been clear at which stage of disease they play a role; early, late or both. To clarify when B lymphocytes are needed, T lymphocytes were transferred from 5-week-old NOD female mice to age-matched NOD/severe combined immunodeficiency (SCID) recipient mice. NOD/SCID mice, which lack functionally mature T and B lymphocytes, do not normally develop insulitis or insulin dependent diabetes melitus (IDDM). The NOD/SCID mice that received purified T lymphocytes from 5-week-old NOD mice subsequently developed insulitis and diabetes even though they did not have detectable B lymphocytes. This suggests that while B lymphocytes may be essential for an initial priming event they are not requisite for disease progression in the NOD mouse. PMID- 11903618 TI - Granulocyte-macrophage colony-stimulating factor activates the transcription of nuclear factor kappa B and induces the expression of nitric oxide synthase in a skin dendritic cell line. AB - Nitric oxide (NO) produced by skin dendritic cells and keratinocytes plays an important role in skin physiology, growth and remodelling. Nitric oxide is also involved in skin inflammatory processes and in modulating antigen presentation (either enhancing or suppressing it). In this study, we found that GM-CSF stimulates the expression of the inducible isoform of nitric oxide synthase (iNOS) in a fetal-skin-derived dendritic cell line (FSDC) and, consequently, increases the nitrite production from 11.9 +/- 3.2 micromol/L (basal level) to 26.9 +/- 4.2 micromol/L. Pyrrolidinedithiocarbamate (PDTC) inhibits nitrite production, with a half maximal inhibitory concentration (IC50) of 19.3 micromol/L and the iNOS protein expression in FSDC. In addition, western blot assays revealed that exposure of FSDC to GM-CSF induces the phosphorylation and degradation of the inhibitor of NF-kappaB (IkB), with subsequent translocation of the p50, p52 and RelB subunits of the transcription nuclear factor kappa B (NF kappaB) from the cytosol to the nucleus. Electrophoretic mobility shift assays (EMSA) showed that FSDC exposure to GM-CSF activates the transcription factor NF kappaB. Together, these results show that GM-CSF induces iNOS expression in skin dendritic cells by a mechanism involving activation of the NF-kappaB pathway. PMID- 11903620 TI - Can MHC class II genes mediate resistance to type 1 diabetes? AB - Numerous studies have associated carriage of HLA-DRB1*1501, DQA1*0102 and DQB1*0602 (DR15, DQ6) with dominant resistance to type 1 diabetes and have concluded that one or more of the component HLA class II molecules mediate this effect. Mechanisms for MHC class II-mediated resistance to diabetes have been proposed from studies of transgenic mice, usually using the diabetes-prone non obese diabetic (NOD) strain. However, these studies have not reached any consensus on a plausible mechanism. In this study we question why the role of central MHC genes in resistance to diabetes has not been addressed, as the central MHC carries markers of susceptibility to diabetes in linkage disequilibrium with several genes with known or putative immunoregulatory functions. To illustrate the type of studies required to address this issue, we selected diabetes patients and control subjects for carriage of HLA-DR15 and the C allele at position +738 in the inhibitor of kappa B-like gene (IKBL). These alleles mark the 7.1 haplotype (HLA-A3, B7, IKBL738*C, DR15, DQ6). HLA-DR15 was the most effective marker of resistance, but an effect may be evident with IKBL738*C in a larger study. Moreover, carriage of the entire haplotype was particularly rare in patients. The best explanation for this is that the critical gene lies between IKBL and HLA-DRB1, and is more closely linked to HLA-DRB1. Candidate genes at the centromeric end of the central MHC are reviewed, highlighting the need for further study. PMID- 11903621 TI - Differential expression of individual gene copies from within a tRNA multigene family in the mulberry silkworm Bombyx mori. AB - In mulberry silkworm Bombyx mori, tRNA1(Gly) constitutes a multigene family from which the individual members are transcribed at different levels in vitro in homologous nuclear extracts. We report here the quantification of functional transcripts of these gene copies in vivo in B. mori-derived BmN cells based on a suppression assay. The gene copies were converted to encode suppressor tRNAs and co-transfected into cell lines with reporter gene(s) harbouring one or more nonsense mutations and the reporter gene activity was quantified. Individual members of the gene family were transcribed to very high-, medium- and very low levels, following the same pattern as in vitro. All these gene copies were maximally expressed in Bm cells as compared to other insect cell lines. PMID- 11903622 TI - Repression of Manduca sexta ferritin synthesis by IRP1/IRE interaction. AB - Mammalian ferritin subunit synthesis is controlled at the translational level by the iron regulatory protein 1 (IRP1)/iron responsive element (IRE) interaction. Insect haemolymph ferritin subunit messages have an IRE in the 5'-untranslated region (UTR). We have shown that recombinant M. sexta IRP1 represses the in vitro translation of both the heavy and light chain ferritin subunits from this species without altering transcription. Deletion of either the 5'-UTR or the IRE from the mRNA abolishes IRP1 repression. Our studies indicated that the translational control of ferritin synthesis by IRP/IRE interaction could occur in insects in a manner similar to that of mammals. To our knowledge, this is the first report of the control of insect ferritin synthesis by IRP1/IRE interaction. Furthermore, this is the first indication that the synthesis of a secreted ferritin subunit can also be controlled in this manner. PMID- 11903623 TI - Functional expression of a locust tyramine receptor in murine erythroleukaemia cells. AB - The LCR/MEL system (Locus Control Region/Murine Erythroleukaemia cells) was employed to express and characterize the Locusta migratoria tyramine receptor (TyrLoc), an insect G protein-coupled receptor. Functional agonist-dependent responses were recorded in stable, tyramine receptor expressing cell clones (MEL TyrLoc). Tyramine elicited a dose-dependent increase of cytosolic Ca2+-ions and an attenuation of forskolin-induced cyclic adenosine monophosphate (AMP) production. Octopamine was shown to be a weak agonist for both responses. In addition, yohimbine proved to be a potent tyramine receptor antagonist. This study reports the first application of the LCR/MEL expression system in functional assays for G protein-coupled receptors and therefore expands the capabilities of this system by exploiting the functionality of the signal transduction pathways. PMID- 11903624 TI - Phylogenetic relationships among fruit flies, Bactrocera (Diptera, Tephritidae), based on the mitochondrial rDNA sequences. AB - Nucleotide sequences of a 1.6 kb long portion of the mitochondrial DNA containing the majority of the 16S rRNA gene, the tRNAval gene, and the 5' half-region of the 12S rRNA gene were determined for forty-eight individuals of nineteen Bactrocera species and one other tephritid taxon, Anastrepha ludens. Phylogenetic analyses were performed using the consistently aligned 1.5 kb long sequences, excluding seventeen portions that could not be aligned unambiguously and were aligned inconsistently among analyses using different programs and parameters. Results of phylogenetic analyses were highly congruent among distance, unweighted parsimony, and weighted parsimony methods in terms of topology of resulting trees. Bootstrap analyses supported many of the phylogenetic relationships resolved by these analyses. Although aligned sequences showed apparent biases in nucleotide content and substitutions that differed among sites within the stem/loop structure of rRNA molecules, compensation for such factors did not greatly affect the topology. The results are discussed in relation to the taxonomic positions of species used in this study and in relation to the phylogenetic and diagnostic utility of the mitochondrial rDNA fragment. PMID- 11903625 TI - Regulation of midgut defensin production in the blood-sucking insect Stomoxys calcitrans. AB - The Stomoxys midgut defensin (Smd) family of genes are exclusively expressed in the anterior midgut of adult flies. Their putative function is protection of the stored bloodmeal from microbial attack. Smd genes are constitutively expressed, up-regulated in response to a bloodmeal and further up-regulated by immune stimulation per os but only in the presence of a bloodmeal not a sugar meal. Smd genes are down-regulated in response to a systemic immune challenge. Smd gene constructs transfected into l(2)mbn cells undertake constitutive expression but are not up-regulated by immune challenge. Electrophoretic mobility shift assays (EMSA) suggest the rel-like sites in the proximal promoter region of Smd genes do not bind midgut factors and so are non-functional. PMID- 11903626 TI - Structure and evolution of the mitochondrial DNA complete control region in the Drosophila subobscura subgroup. AB - The complete A + T-rich region of mitochondrial DNA (mtDNA) has been cloned and sequenced in the species of the Drosophila subobscura subgroup D. subobscura, D. madeirensis and D. guanche. Comparative analysis of these sequences with others already published has identified new sequence motifs that are conserved in Drosophila and other insects. A putative bi-directional promoter and a stop signal are proposed to be involved in the primary mtDNA strand replication of Drosophila. This region strongly resolves relationships of the species included in a phylogenetic analysis, both for closely related species and also at deeper phylogenetic levels when only the left and central domains are taken into account. PMID- 11903627 TI - A DNA fingerprinting procedure for ultra high-throughput genetic analysis of insects. AB - Existing procedures for the generation of polymorphic DNA markers are not optimal for insect studies in which the organisms are often tiny and background molecular information is often non-existent. We have used a new high throughput DNA marker generation protocol called randomly amplified DNA fingerprints (RAF) to analyse the genetic variability in three separate strains of the stored grain pest, Rhyzopertha dominica. This protocol is quick, robust and reliable even though it requires minimal sample preparation, minute amounts of DNA and no prior molecular analysis of the organism. Arbitrarily selected oligonucleotide primers routinely produced approximately 50 scoreable polymorphic DNA markers, between individuals of three independent field isolates of R. dominica. Multivariate cluster analysis using forty-nine arbitrarily selected polymorphisms generated from a single primer reliably separated individuals into three clades corresponding to their geographical origin. The resulting clades were quite distinct, with an average genetic difference of 37.5 +/- 6.0% between clades and of 21.0 +/- 7.1% between individuals within clades. As a prelude to future gene mapping efforts, we have also assessed the performance of RAF under conditions commonly used in gene mapping. In this analysis, fingerprints from pooled DNA samples accurately and reproducibly reflected RAF profiles obtained from individual DNA samples that had been combined to create the bulked samples. PMID- 11903628 TI - Recurrent gains and losses of large (84-109 bp) repeats in the rDNA internal transcribed spacer 2 (ITS2) of rhipicephaline ticks. AB - We studied the internal transcribed spacer 2 (ITS2) in twenty-two spp. of ticks from the subfamily Rhipicephalinae. A 104-109 base pair (bp) region was imperfectly repeated in most ticks studied. Mapping the number of repeat copies on to a phylogeny from the ITS2 showed that there have been many independent gains and losses of repeats. Comparison of the sequences of the repeat copies indicated that in most taxa concerted evolution had played little if any role in the evolution of these regions, as the copies clustered by sequence position rather than species. In our putative secondary structure, each repeat copy can fold into a distinct and almost identical stem-loop complex; a gain or loss of a repeat copy apparently does not impair the function of the ITS2 in these ticks. PMID- 11903629 TI - Germline transformation of the malaria vector, Anopheles gambiae, with the piggyBac transposable element. AB - Germline transformation of the major African malaria vector, Anopheles gambiae, was achieved using the piggyBac transposable element marked with the enhanced green fluorescent protein (EGFP) injected into mosquito embryos. Two G1 generation male mosquitoes expressing EGFP were identified among 34 143 larvae screened. Genomic Southern data and sequencing of the piggyBac insertion boundaries showed that these two males arose from one piggyBac insertion event in the injected G0 embryos. Genetic cross data suggest that the insertion site of the element either resulted in, or is tightly linked to, a recessive lethal. This was demonstrated by a deficiency in the number of EGFP-expressing offspring from inbred crosses but expected ratios in outcrosses to non-transformed individuals and failure to establish a pure-breeding line. The insertion was weakly linked to the collarless locus on chromosome 2 and was shown by in situ hybridization to be located in division 28D of that chromosome. Particularly high levels of expression were observed uniformly in salivary glands and, in most individuals, in the anterior stomach. An improvement in the injection technique at the end of the studies resulted in increased G0 hatching, transient expression and EGFP expression rates among G1 progeny. PMID- 11903630 TI - Fine scale mapping in the sex locus region of the honey bee (Apis mellifera). AB - Isolating an unknown gene with fine-scale mapping is possible in a "non-model" organism. Sex determination in honey bees consists of a single locus (sex locus) with several complementary alleles. Diploid females are heterozygous at the sex locus, whereas haploid males arise from unfertilized eggs and are hemizygous. The construction of specific inbred crosses facilitates fine scale mapping in the sex locus region of the honey bee. The high recombination rate in the honey bee reduces the physical distance between markers compared with model organisms and facilitates a novel gene isolation strategy based on step-wise creation of new markers within small physical distances. We show that distances less than 25 kb can be efficiently mapped with a mapping population of only 1000 individuals. The procedure described here will accelerate the mapping, analysis and isolation of honey bee genes. PMID- 11903631 TI - Overproduction of a P450 that metabolizes diazinon is linked to a loss-of function in the chromosome 2 ali-esterase (MdalphaE7) gene in resistant house flies. AB - Up-regulation of detoxifying enzymes in insecticide-resistant strains of the house fly is a common mechanism for metabolic resistance. However, the molecular basis of this increased insecticide metabolism is not well understood. In the multiresistant Rutgers strain, several cytochromes P450 and glutathione S transferases are constitutively overexpressed at the transcriptional level. Overexpression is the result of trans-regulation, and a regulatory gene has been located on chromosome 2. A Gly137 to Asp point mutation in alphaE7 esterase gene, leading to the loss of carboxylesterase activity, has been associated with organophosphate resistance in the house fly and the sheep blowfly. We show here that purified recombinant CYP6A1 is able to detoxify diazinon with a high efficiency. We also show that either the Gly137 to Asp point mutation in alphaE7 esterase gene or a deletion at this locus confer resistance and overproduction of the CYP6A1 protein. Based on these findings, we propose it is the absence of the wild-type Gly137 allele of the alphaE7 gene that releases the transcriptional repression of genes coding for detoxification enzymes such as CYP6A1, thereby leading to metabolic resistance to diazinon. PMID- 11903632 TI - Developmental variation in epidermal growth factor receptor size and localization in the malaria mosquito, Anopheles gambiae. AB - The AGER gene encoding the epidermal growth factor receptor (EGFR) of the malaria mosquito Anopheles gambiae was cloned and sequenced. It represents a canonical member of this family of tyrosine kinase proteins exhibiting many similarities to orthologues from other species, both on the level of genomic organization and protein structure. The mRNA can be detected throughout development. Western analysis with an antibody raised against the extracellular domain of the mosquito protein suggests developmental variation in protein size and location that may be involved in the function of EGFR in the mosquito. PMID- 11903655 TI - The effects of oxytocin and arginine vasopressin in vitro on epididymal contractility in the rat. AB - Oxytocin (OT) and arginine vasopressin (AVP) have been found to be present in the reproductive system of the mammalian male, but their function in this system is unclear. This study examined the effects of these two hormones on contractions of the rat caput epididymis, as well as the hormones' effects on epididymal diameter during contractions. The initial segments of caput epididymides were observed in vitro in a solution of modified Tyrode's solution, with a test solution containing a concentration of either OT or AVP being added after a 5-min period. The frequency and diameter of the epididymal contractions were measured before and after addition of each test solution. Also observed was the effect of magnesium concentrations on the ability of OT to induce changes in epididymal contractility. This study found that the frequency of epididymal contractions increase and that the diameter of the epididymal tubules decrease proportionally during contractions with the addition of either OT or AVP. Furthermore, we noted that Mg++ has an inverse correlation with the frequency of contractions of the caput epididymis both in the presence and absence of OT. These findings suggest that both OT and AVP may have a role in regulating epididymal contractility and thus, perhaps, sperm transport through that organ. PMID- 11903656 TI - Enzymatic and immunochemical evaluation of phospholipid hydroperoxide glutathione peroxidase (PHGPx) in testes and epididymal spermatozoa of rats of different ages. AB - Selenium (Se) and selenoproteins such as glutathione peroxidases are necessary for the proper development and fertilizing capacity of sperm cells. Phospholipid hydroperoxide glutathione peroxidase (PHGPx, E.C. 1.11.1.12) is a monomeric seleno-enzyme present in different mammalian tissues in soluble and bound form. Its function, like the other glutathione peroxidases, was originally viewed as a protective role against hydroperoxides, but direct and indirect evidence indicates that it has additional regulatory roles. PHGPx is present in testis cells and sperm cells, and its appearance is hormone regulated. We present here biochemical data, which clearly indicate that the enzyme specific activity in rat is age-dependent during the life-span monitored (from 36 to 365 days), with a maximum at 3 months of age in the testis germ cells and at 6 months of age in the isolated epididymal sperm cells. Western blotting and immunocytochemical analysis by means of anti-PHGPx antibodies show the different distribution and the strong binding of PHGPx in the testes and sperm cell subcellular compartments (nucleus, acrosome, mitochondria and residual bodies) of rats of different age. The presence of the protein exhibits in the testis cells a pattern different from that of the catalytic activity, with a maximum at 6 months of age. The subcellular distribution of PHGPx is qualitatively, but not quantitatively, unchanged during ageing. These different behaviours are compared and discussed. PMID- 11903657 TI - Capacitation status and in vitro fertility of boar spermatozoa: effects of seminal plasma, cumulus-oocyte-complexes-conditioned medium and hyaluronan. AB - In the present study, the effects of seminal plasma (SP), cumulus-oocyte complexes (COCs) conditioned medium (CCM) and hyaluronan (HA) on functional changes and in vitro fertilizing ability of porcine spermatozoa were examined. In in vitro fertilization (IVF) experiments, 10% (v/v) of exogenous SP in the fertilization medium prevented sperm penetration (using fresh-extended and frozen thawed ejaculated spermatozoa). Analysis of frozen-thawed CCM revealed a HA content to levels of 30 ng/mL per incubated COC. Presence of frozen-thawed CCM did not, however, prove effective to increase (furthermore decreasing) oocyte penetration in vitro, and neither did supplementation with exogenous HA at the same concentration as that present in the CCM (secreted by COCs). Analysis of sperm capacitation using the chlortetracycline (CTC) assay showed that frozen thawed CCM had no elevating effect on 'B-pattern' spermatozoa (implying capacitation-like changes) and that addition of 10% (v/v) SP held spermatozoa in the 'F-pattern' (intact) status. Dose of 500 microg/mL HA and freshly prepared CCM increased, however, the frequency of capacitated spermatozoa (B-pattern) without resulting in increased rates of 'AR-pattern' (acrosome-reacted) spermatozoa, compared with controls. The present results confirm the decapacitating effect of SP and suggest capacitating actions of HA (dose-related) and CCM (freshly prepared) on boar spermatozoa in vitro. The unclear effects of frozen-thawed CCM and a low dose of HA on penetration rates of boar spermatozoa call for further researches of their function in vivo. PMID- 11903658 TI - The effect of (R,S)-ornidazole on the fertility of male mice and the excretion and metabolism of 36Cl-(R,S)-ornidazole and 36Cl-(R,S)-alpha-chlorohydrin in male mice and rats. AB - (R,S)-Ornidazole, an effective antifertility agent for male rats at 400 mg/kg/day, was ineffective at this dose in male mice and at 1000 mg/kg/day caused neural effects. The compound was not excreted unchanged and more polar metabolites and Cl- were detected in 0-8 h urine following a single injection (400 mg/kg). In 8-24 h urine even these metabolites and most Cl ion were absent, indicating rapid metabolism of ornidazole. There was no organ specific accumulation of 36Cl-(R,S)-ornidazole in murine tissues. After injection of 36Cl (R,S)-alpha-chlorohydrin, another antifertility agent in the rat but not the mouse, there was also no tissue-specific accumulation of radioactivity in the reproductive tract of either species. Urinary excretion rates of alpha chlorohydrin were twice as rapid in mice as in rats. In mice, alpha-chlorohydrin was the major urinary metabolite, but in the rat metabolites included Cl-, 3 chlorolactate (BCLA) at 5 and 10 h and BCLA only at 24 h. BCLA was the major metabolite detected in most tissues at 10 and 24 h. In the rat cauda (but not caput) epididymidis the glycolytic inhibitor 3-chlorolactaldehyde was present at 5 h (but not 10 h), indicative of early metabolism. These results demonstrate a greater metabolism and excretion of putative antifertility agents in the mouse than the rat, lowering the amount of effective inhibitor circulating in the animal, which may explain why (R,S)-alpha-chlorohydrin and (R,S)-ornidazole are ineffective in this species at the dosages and injection times used, despite their spermatozoa being sensitive to inhibition by (R,S)-alpha-chlorohydrin in vitro. PMID- 11903659 TI - Application of a 1.48-microm diode laser for bisecting oocytes into two identical hemizonae for the hemizona assay. AB - Laser systems are very promising new technical tools in assisted reproduction. It was investigated if laser radiation can replace the mechanical cutting procedure via micromanipulator in the hemizona assay (HZA), a commonly used bioassay to determine the sperm-zona pellucida binding capacity. An oocyte was bisected precisely into two identical hemizonae with approximately 20 laser pulses (pulse length 30 msec) using a 1.48-microm diode laser. Compared with the conventional method using microscalpels for zona bisection, laser treated hemizonae showed equivalent sperm-binding and within the two groups there was no detectable difference between matching hemizonae in their capacity for tight sperm-binding. To evaluate whether laser radiation affects the outcome of the HZA when effects of certain substances are investigated, the spermatozoa were preincubated with human follicular fluid (hFF), which inhibits the binding of spermatozoa to zona pellucida in vitro. Supplementation with follicular fluid exerted an inhibitory effect in both groups. The hemizona index (HZI) showed no statistical differences between the two methods. Therefore, the 1.48-microm diode laser is a suitable new instrument for generating equally sized hemizonae. There is no use for holding pipettes and microscalpels, on the contrary, for performing the HZA the laser is a precise, very quick and easy to use new working tool. PMID- 11903660 TI - Effect of human sperm exposure to progesterone on sperm-oocyte fusion and sperm zona pellucida binding under various experimental conditions. AB - In this study, the effect of human sperm exposure to progesterone on sperm/oocyte fusion, using the hamster egg penetration test, and on sperm/zona pellucida (ZP) binding, using the hemizona assay, was investigated under various experimental conditions. A brief exposure of human spermatozoa to progesterone exerted a stimulatory effect on sperm/oocyte fusion which was dose-dependent, capacitation dependent, influenced by the source of serum albumin in capacitating medium, and was higher than that produced by the exposure to progesterone from the onset of capacitation. The exposure of capacitated spermatozoa to progesterone during 20 min-spermatozoa/ZP-coincubation produced an enhancement of ZP-binding, which was not significantly influenced by the source of serum albumin in capacitating medium. A significantly lower ZP-binding was exhibited by spermatozoa exposed to progesterone from the beginning of capacitation. These results indicate that progesterone exerts a stimulatory effect on human sperm's fertilizing ability, which occurs mainly in post-capacitation events directly involved in sperm/oocyte fusion and in ZP-binding. Conditions optimizing these effects are provided. They should be taken into account in the standardization of experimental and clinical studies designed to evaluate the response of human spermatozoa to progesterone. PMID- 11903661 TI - Dietary fatty acid composition affects aminopeptidase activities in the testes of mice. AB - The autocrine/paracrine control mechanisms of local factors, such as the renin angiotensin system and the thyrotropin-releasing hormone (TRH), seem to play a relevant role in testicular physiology. It has been proposed that dietary fat composition influences male reproductive function modifying the cholesterol phospholipid composition of testicular plasma membranes. Modifications in the composition and physical properties of the membranes may lead to alterations in the activities of membrane-bound (M-B) enzymes. We have previously demonstrated that cholesterol and steroid hormones affect aminopeptidase (AP) activities. Dietary fatty acids with different degrees of saturation modified AP activities in the serum of mice and an olive oil supplemented diet influenced the AP activities in the testes of mice. We hypothesized that the modification of dietary fat composition may affect angiotensin- [glutamyl-AP (GluAP), aspartyl-AP (AspAP)] and TRH- [pyroglutamyl-AP (pGluAP)] degrading activities in the testis. In this study, we investigated the effect of diets supplemented with sunflower oil (SFO), fish oil (FO), olive oil (OO), lard (L) or coconut oil (CO) on soluble (Sol) and M-B GluAP, AspAP and pGluAP in mice testis, using arylamides as substrates. Sol GluAP activity did not show differences among groups. However, Sol AspAP and Sol pGluAP progressively decreased with the degree of saturation of the fatty acid used in the diet. In contrast, M-B GluAP progressively increased with the degree of saturation of the fatty acid used in the diet. For M-B AspAP activity, mice fed diets containing FO showed significantly higher levels than those fed diets containing SFO, OO and L but not those containing CO. For M-B pGluAP activity, the highest levels were observed for mice fed diets containing FO and OO. The present data suggest that the type of fat used in the diet may influence the autocrine/paracrine functions of locally synthesized angiotensin peptides and TRH in the testis, and consequently may be important in male reproductive functions. PMID- 11903662 TI - Androgen administration in middle-aged and ageing men: effects of oral testosterone undecanoate on dihydrotestosterone, oestradiol and prostate volume. AB - The gradual reduction of plasma testosterone in middle-aged and older men from mid-life onwards coincides paradoxically with the time when there is progressive growth of the prostate, a highly androgen-dependent organ. The growing interest in androgen therapy for older men makes it essential to understand the effects of exogenous testosterone on the non-diseased prostate, yet few studies are available. The present study examined prostate volume, prostate-specific antigen (PSA) and lower urinary tract symptom (IPSS) score in 207 men, aged 40-83 years, presenting with clinical features of age-related androgen deficiency [sexual and/or urinary dysfunction, elevated lutenizing hormone (LH)] who were treated for 6 months with oral testosterone undecanoate (TU). Men were divided into two groups, group 1 (n=92, plasma testosterone levels > 13 nmol/L) were treated with 80 mg daily; group 2 (n=115, plasma testosterone levels < 13 nmol/L) were treated with given 120 mg daily. Before treatment and after 1, 3 and 6 months of treatment, prostate volume was measured by ultrasound and hormones [testosterone, dihydrotestosterone, oestradiol, LH, follicle-stimulating hormone (FSH)] and PSA were measured. Within 1 month of treatment, the elevated blood LH levels were markedly decreased in all men in group 1, as well as most men in group 2. Group 2 was subdivided into men whose LH levels were suppressed (n=95, group 2a) and those whose LH levels did not suppress (n=20, group 2b). Men in group 1 and 2a had marked decreases in prostate volume, PSA and lower urinary tract symptom (IPSS) scores whereas no significant changes were observed in group 2b. Groups 1 and 2a also had more striking suppression of LH, FSH, dihydrotestosterone and oestradiol whereas group 2b had no significant increases in blood testosterone concentrations. These findings suggest that exogenous testosterone in middle-aged and older men with some clinical features of age-related androgen deficiency can retard or reverse prostate growth and that elevated plasma LH may be a useful index of severity of age-related androgen deficiency. PMID- 11903665 TI - Sporotrichosis. PMID- 11903666 TI - Factitious disease of the breast of a male due to injection of liquid plastic. AB - BACKGROUND: Factitious disease involving the breast is unusual. The rarely reported cases of this entity have been encountered in middle-aged women. In general, factitious disease can be distinguished on clinical and/or psychological grounds from self-induced disorders seen in malingerers and among those demonstrating Munchausen's syndrome. METHODS: We report herein a male patient displaying factitious disease of the breast due to injection of a high viscosity liquid plastic material. RESULTS: Establishment of the proper diagnosis was greatly delayed due to a lack of suspicion of this entity. Only direct confrontation of the patient with the biopsy results (lipogranulomatosis) led to a reluctant and then only partial admission of the self-induced nature of this patient's illness. CONCLUSION: The clinician should remain vigilant for factitious disease when confronted with chronic or recurrent lesions of a bizarre or atypical morphology. Any body site can be involved, including the breast. Management is difficult and is best accomplished in conjunction with an appropriate mental health professional. Continued dramatic surgical interventions are contraindicated. PMID- 11903667 TI - Expression of bcl-2 protein in active skin lesions of Behcet's disease. AB - BACKGROUND: Expression of bcl-2 protein has been shown to play an important role in the pathogenesis of some inflammatory as well as neoplastic disorders. In this study we have investigated the presence of bcl-2 protein in active skin lesions of Behcet's disease and compared these results with normal skin samples of Behcet's disease (BD) patients and BD unrelated leukocytoclastic vasculitis. METHODS: Active skin lesions of 23 Behcet's disease patients, normal skin samples of seven Behcet's disease patients, and archival biopsy specimens of 23 cutaneous leukocytoclastic vasculitis were investigated for the presence of bcl-2 protein by immunohistochemical methods. Results of staining were assessed semiquantitatively. Chi-square tests were used for statistical analysis. RESULTS: Expression of bcl-2 protein were demonstrated in 16 of 23 (69.5%) and 8 of 23 (34.7%) patients with Behcet's disease and leukocytoclastic vasculitis, respectively. There were statistically significant difference between two groups (x2 = 4.27, P < 0.05). None of the normal skin samples of Behcet's disease patients showed bcl-2 expression. CONCLUSION: Expression of bcl-2 protein may play a particular role in the development of skin lesions in Behcet's disease by causing prolonged survival of infiltrating lymphocytes. PMID- 11903668 TI - Pancreatic panniculitis in a kidney transplant recipient. AB - BACKGROUND: Pancreatic panniculitis is a rare complication of pancreatic disease. Histopathologic findings are pathognomonic and may be helpful for early diagnosis. METHODS: We present a female kidney transplant recipient, with systemic lupus erythematosus on immunossuppresive therapy, who developed panniculitis. RESULTS: Histological features strongly suggested pancreatic panniculitis and this was confirmed by clinical, laboratory and image findings. CONCLUSION: The characteristic histologic features of pancreatic panniculitis may help to uncover undiagnosed pancreatic disease. Early therapy may avoid ensuing serious complications. PMID- 11903669 TI - Allergic contact dermatitis to temporary tattoos with positive para phenylenediamine reactions: report of four cases. AB - BACKGROUND: In recent years, temporary tattoos instead of permanent tattoos have become popular worldwide. Although contact allergy to temporary henna tattoos appears to be rare in the past, it is progressively more commonly reported. PATIENTS: Four Taiwanese patients of allergic contact dermatitis following application of temporary tattoos were patch tested and they were followed up for 1 year after treatment. RESULTS: All four of the tested patients were positive to paraphenylenediamine. At 1-year follow-up, all four patients still showed various degrees of remnant hyperpigmentation on their previous tattooed areas. CONCLUSION: Temporary tattoos may pose similar risks of allergic reactions associated with permanent tattoos. A high risk of prolonged post-inflammatory hyperpigmentation after allergic contact dermatitis from temporary tattoos should also be alerted, especially in Asian skin type. PMID- 11903670 TI - Lichen planus pemphigoides: report of two cases. PMID- 11903671 TI - A case of cutaneous lymphangiectasis secondary to breast cancer treatment. PMID- 11903672 TI - Oral erosive lichen planus associated with thymoma. PMID- 11903673 TI - A case of mucosal leishmaniasis: beneficial usage of polymerase chain reaction for diagnosis. PMID- 11903674 TI - Lichenoid photodermatitis associated with nimesulide. PMID- 11903675 TI - Mycobacterium chelonae/abscessus complex infection in a liver transplant patient. PMID- 11903676 TI - Generalized lichenoid drug eruption following Blaschko lines. PMID- 11903677 TI - Eccrine syringosquamous metaplasia. PMID- 11903678 TI - Cutaneous metastases from hepatocellular carcinoma. PMID- 11903680 TI - The Philippine Society of Cutaneous Medicine's 16th annual convention "Dermatology: insights and innovations". PMID- 11903679 TI - A simple surgical technique for the treatment of steatocystoma multiplex. AB - BACKGROUND: Since steatocystoma multiplex is an uncommon disorder, its treatment has not been discussed in detail in the textbooks and surgical excision is the most commonly mentioned method. In this article, we describe a very simple surgical technique, which was developed by modifying previous reports. METHODS: A 29-year-old woman, diagnosed clinically and histologically as steatocystoma multiplex, was treated with this modified technique. We punctured the cysts under local anesthesia with a sharp-tipped cautery point and evacuated the contents by squeezing the cyst with a fine forceps. Then, the cyst wall was grasped by the forceps and the sacs were extracted through small holes. More than 50 cysts were treated. RESULTS: The treatment was well tolerated by the patient. No complications developed during or after the procedure. After 14 months follow up no recurrences were observed and the results were cosmetically excellent. CONCLUSIONS: This modified technique is very simple and time saving. Its cosmetic and long-term results are successful. We believe that it must be considered as the treatment of choice for steatocystoma multiplex. PMID- 11903681 TI - Synopsis of the 15th Chinese Congress of Dermatopathology. PMID- 11903682 TI - Sweet's syndrome associated with francisella tularensis infection. PMID- 11903683 TI - Value of prostate specific antigen alpha1-antichymotrypsin complex for the detection of prostate cancer in patients with a PSA level of 4.1-10.0 ng/mL: comparison with PSA-related parameters. AB - BACKGROUND: The aim of the present study was to evaluate the usefulness of prostate specific antigen alpha1-antichymotrypsin complex (PSA-ACT) in the differential diagnosis of prostate cancer in patients with a PSA level of 4.1 10.0 ng/mL compared to several PSA- and PSA-ACT-related parameters. METHODS: Serum samples were obtained from 103 patients with no evidence of malignancy on biopsy and 29 with histologically confirmed prostate cancer. All patients had pretreatment serum PSA levels between 4.0 and 10.0 ng/mL. The different forms of serum PSA, including total PSA (tPSA), free PSA (fPSA) and PSA-ACT were measured using immunofluorometric techniques with different monoclonal antibodies against PSA and ACT. Furthermore, tPSA and PSA-ACT densities of the whole prostate (PSAD and ACTD, respectively) and the f-to-tPSA and the f-to-PSA-ACT ratios (F/T ratio and F/ACT ratio, respectively) were calculated. RESULTS: The differences between patients with prostate cancer and benign prostatic disease were significant with respect to all six parameters examined in this study. Analysis of receiver operating characteristics revealed that the areas under the curve for PSA-ACT, ACTD and the F/ACT ratio were larger than those for tPSA, PSAD and the F/T ratio, respectively. However, there were no significant differences in discrimination between benign and malignant diseases among these six parameters. CONCLUSIONS: In patients who have an intermediate serum PSA level, PSA-ACT and its associated parameters may not be significantly superior in the differential diagnosis between prostate cancer and benign prostatic diseases compared to tPSA and its traditional relatives. PMID- 11903684 TI - The correlation between penile tumescence measured by the erectometer and penile rigidity by the RigiScan. AB - BACKGROUND: The goal of this study was to evaluate the correlation between increments of penile tumescence and penile rigidity measured by the erectometer and the RigiScan, respectively. METHODS: Nocturnal penile tumescence (NPT) was measured in 25 volunteers (mean age, 49.5 years). The erectometer and the RigiScan were used simultaneously for a total of 47 nights. We studied the correlation between maximum penile circumferential changes determined by the erectometer and penile rigidity patterns measured by the RigiScan. RESULTS: Maximum circumferential changes during NPT measured by the erectometer were well correlated to those determined by the RigiScan (correlation coefficient, 0.719). In addition, penile circumferential changes measured by the erectometer corresponded well to the penile rigidity pattern determined by the RigiScan (P=0.0001). Specifically, maximum penile circumferential changes of more than 30 mm and less than 10 mm had 70% positive predictive value and 100% negative predictive value for predicting the normal rigidity pattern, respectively. CONCLUSIONS: Penile circumferential changes measured by the erectometer were well correlated to penile rigidity measured by the RigiScan, particularly when the increments were larger than 30 mm or less than 10 mm. These results suggested that the erectometer was a useful tool to estimate the penile rigidity patterns of the RigiScan. PMID- 11903685 TI - Need for sling surgery in patients with large cystoceles and masked stress urinary incontinence. AB - BACKGROUND: We studied the need for sling surgery in patients who suffered from large cystoceles and masked stress urinary incontinence. METHODS: Twenty patients who had large cystoceles but neither evidence nor history of stress incontinence were enrolled in this study. The cystocele was reduced using a reducing device. Masked urinary incontinence was identified by a 60 m pad weighing test and a stress test. The cystocele was reduced using a pessary ring in 14 patients, or a vaginal pack formed of two rolls of ordinary 28 x 28 cm gauze in six patients. Ten of 20 patients were diagnosed with masked stress urinary incontinence and were treated with anterior colporrhaphy and a suburethral sling procedure. The other 10 patients were continent after use of a cystocele reducing device and were treated with anterior colporrhaphy alone. Average follow-up periods of the patients with or without masked stress urinary incontinence were 51.2 months (range, 24.0-72.0 months) or 57.6 months (range, 27.0-70.0 months), respectively. RESULTS: One of the 10 patients diagnosed with masked stress incontinence had mild stress urinary incontinence postoperatively. None of the 10 continent patients had stress incontinence after anterior colporrhaphy alone. CONCLUSIONS: Reducing devices of protruding cystocele were clinically useful in the detection of masked stress incontinence. Sling surgery was effective to prevent emerging stress urinary incontinence for patients who suffered from cystocele and masked stress incontinence. PMID- 11903686 TI - Changes in cellular immunity during chemotherapy for testicular cancer. AB - BACKGROUND: The changes in vivo in immunocyte functions during chemotherapy that is administered in combination with granulocyte colony-stimulating factor (G-CSF) in humans have not been fully investigated. This study was designed to examine neutrophil functions and the activities of natural killer (NK) cells, during the administration of chemotherapy and G-CSF for the treatment of testicular cancer. METHODS: Seven patients with germ cell tumors at stage IIA, IIB or IIIB, who were treated with bleomycin, etoposide and cisplatin (BEP), were enrolled in the study. Numbers and activities of neutrophils and NK cells were measured at various times during and after the first course of chemotherapy. Neutrophil phagocytosis was quantitated by flow cytometry with fluorescent latex beads. Bactericidal activity was measured in terms of colony-forming units. The activity of NK cells was measured by monitoring the release of 51Cr. RESULTS: After BEP chemotherapy, CD16+ and CD56+ cell counts, and neutrophil granulocyte counts decreased while there were no significant changes in the number of lymphocytes. Phagocytosis by neutrophils was enhanced after administration of G-CSF. The activity of NK cells was severely impaired after chemotherapy and did not change after administration of G-CSF. CONCLUSIONS: After BEP chemotherapy for testicular cancer with G-CSF, neutrophil function was not at all inferior to those before treatment. Natural killer cell activity was suppressed by the BEP chemotherapy and did not change after administration of G-CSF. PMID- 11903687 TI - Growth fractions of human renal cell carcinoma defined by monoclonal antibody Ki 67. Predictive values for prognosis. AB - BACKGROUND: We used immunohistochemical techniques to elucidate the role of growth fractions of renal cell carcinoma in the clinicopathology of the condition and patient survival. METHODS: Fifty-two fresh-frozen nephrectomy specimens were immunostained with Ki-67 monoclonal antibody. Ki-67 indexes were determined to examine the relationship between tumor size, grade, stage and survival curve. This study included 43 men and nine women with the mean age 58.4 +/- 11.7 years, who had been followed up for 39 +/- 25 months. RESULTS: The Ki-67 index ranged from 0.6 to 14.1%, averaging 4.6 +/- 5.8%. It was 2.8 +/- 2.4% in tumors <5 cm, 4.7 +/- 3.6% in tumors > or =5 cm and 7.1 +/- 9.0% in tumors > or =10 cm. The Ki 67 index of grades 1, 2 and 3 tumors was 2.3 +/- 1.1%, 3.3 +/- 2.7% and 12.0 +/- 10.4%, respectively. Grade 3 tumors had a significantly higher Ki-67 index than grade 1 or grade 2 tumors. There was no correlation between the Ki-67 index and tumor stage. Patients with a Ki-67 index < 5.6% had a better prognosis than those with an index > 5.6% (P=0.029). However, multivariate analysis demonstrated that tumor size (P=0.034) and grade (P=0.038) were higher in hazard ratio than the Ki 67 index. CONCLUSIONS: Most renal cell carcinomas had low growth fractions. Although a high Ki-67 index should indicate a poor prognosis, Ki-67 did not correlate to metastasis. We believe it is necessary to investigate the factors, other than growth potential, that affect metastasis. PMID- 11903689 TI - Effects of quercetin on the heat-induced cytotoxicity of prostate cancer cells. AB - BACKGROUND: It has been demonstrated that prostate cancer cells are relatively sensitive to heat stress. We have reported that heat treatment at 43 degrees C increases the expression of heat shock protein 70 (hsp70) in prostate cancer cells, leading to apoptosis. Hsp70 is a protein that protects cells against heat damage. Cells with lower levels of hsp70 have been shown to have a higher sensitivity to heat stress. Therefore, downregulation of hsp70 is expected to enhance heat-induced inhibitory effects on cell growth. Quercetin has been reported to be an agent that inhibits hsp70 expression. The present study was undertaken to investigate the effects of quercetin and/or heat on the growth of prostate cancer cells in vitro. METHODS: Three human prostate cancer cell lines were used: Lncap; PC-3; and JCA-1. The cells were treated with quercetin and/or heat. Alterations in the cell cycle and hsp70 expression were examined by means of flow cytometry (FCM). The apoptotic cells were detected by FCM using fluorescein isothiocyanate (FITC) labeled annexin V. RESULTS: Treatment with quercetin alone resulted in an apparent decrease of hsp70-positive cells and an increase of subG1 cells in JCA-1 and LNcap cells. Quercetin inhibited an increase of hsp70 expression after heat treatment and increased the number of subG1 cells with lower levels of hsp70 in JCA-1 and LNcap cells. Quercetin was found to enhance heat-induced inhibitory effects on cell growth and heat-induced apoptosis in both JCA-1 and LNcap cells. CONCLUSION: These results suggest that quercetin may enhance heat-induced cytotoxicity in prostate cancer cell lines through the inhibition of hsp70 production. PMID- 11903688 TI - GnRH analog, leuprorelin acetate, promotes regeneration of rat spermatogenesis after severe chemical damage. AB - BACKGROUND: Future fertility is a major concern for cancer patients who undergo intensive chemotherapy. There has been controversy about whether hormonal treatments may have protective effects against the severe spermatogenic damage caused by chemotherapy or irradiation. Recently, it has been proposed that gonadotrophin-releasing hormone (GnRH) analogs administered after testicular damage stimulate the recovery of spermatogenesis. In this study, we have investigated the effects of GnRH agonist, leuprorelin, on the damage to spermatogenesis induced by busulfan. METHODS: Fisher rats were treated with busulfan, 25 mg/kg, intraperitoneally. The effects of subcutaneous injections of leuprorelin before or after treatment were evaluated histologically 18 weeks later. RESULTS: The percentage of 'recovered' seminiferous tubules was 27.7 +/- 12.6% in control rats without leuprorelin and 26.9 +/- 10.2% in rats with leuprorelin injected 4 weeks before busulfan. Rats in both groups showed poor recovery of spermatogenesis with an increase of intratesticular fluid. However, rats treated with leuprorelin three times (4 weeks apart) after busulfan showed an improvement of up to 56.5 +/- 12.0% (P < 0.05). A focal but massive necrotic lesion in the testis was observed only in this group of rats. CONCLUSIONS: The results demonstrated that leuprorelin administered after chemical testicular damage enhanced the recovery of spermatogenesis. At the same time, a possible significant side-effect of leuprorelin was noted. PMID- 11903690 TI - Asynchronous (heterochronic) bilateral renal infarction associated with primary antiphospholipid syndrome. AB - We report the rare case of a 51-year-old man with asynchronous (heterochronic) bilateral renal infarction associated with primary antiphospholipid syndrome. He was treated for right renal infarction, but 2 months later, while under anticoagulant treatment, he had a recurrence on the other side of the renal infarction. The laboratory work-up confirmed antiphospholipid syndrome. Six months later the patient has not experienced any new thrombotic episodes and is receiving oral anticoagulants and antiplatelet therapy. PMID- 11903691 TI - Extraperitoneal bladder rupture secondary to rectal impalement. AB - Impalement injury to both the urinary bladder and the rectum is a rare occurrence. A 45-year-old man was referred to our hospital because he had been stabbed through the internal femoral skin by a steel bar. Retrograde cystography and computed tomography failed to demonstrate bladder rupture. We carried out cystoscopy (CS) because of the existence of gross hematuria and found a penetrating wound in the posterior wall. Colon fiberscopy revealed two wounds in the rectum, one of which communicated with the bladder. A diagnostic laparoscopy revealed no associated injuries in the peritoneal cavity. Vesicorectal injury was diagnosed and treatment included transanal closure of fistulas and indwelling ureteral and urethral catheters. Postoperative CS revealed complete closure of the injury. PMID- 11903692 TI - Retroperitoneal cystic metastasis from a small clear cell renal carcinoma. AB - A 39-year-old housewife was referred to our hospital for the treatment of a small renal tumor. A 25 x 35 mm cystic mass that had been detected by computerized tomography scan just caudal to the renal hilus proved to be a metastasis from the renal carcinoma of clear cell type. The pathogenesis may have been due to tumor cells obstructing a lymphatic vessel draining the kidney. Cystic metastasis from renal cell carcinoma is very rare and this appears to be the second published case in the world. PMID- 11903693 TI - Mycobacterium tuberculosis infection of bilateral cervical lymph nodes after renal transplantation. AB - We report the case of a 52-year-old man who underwent a renal transplantation and subsequently developed extrapulmonary tuberculosis. The immunosuppressive agent was intravenously administered continuously together with antituberculosis drugs. The tuberculosis improved and renal function has been well preserved for more than 3 years post transplantation. PMID- 11903694 TI - Early stage small cell carcinoma of the urinary bladder. AB - A 73-year-old man with primary small cell carcinoma of the bladder underwent radical cystectomy. The pathological findings revealed the tumor confined to the submucosal layer (pT1) without metastasis. No adjuvant chemotherapy was carried out. He is alive with no evidence of the disease 24 months after the operation. PMID- 11903695 TI - Neurofibromatosis involving the urinary bladder. AB - We present two interesting cases of a 24-year-old man and a 14-year-old boy, uncle and nephew, with lower urinary tract symptoms, cafe au lait patches and subcutaneous nodules. Ultrasonography and computed tomography scans showed a large, irregular lobulated soft tissue mass between the bladder and sacrum. Cystoscopy, laparotomy and biopsies revealed neurofibromatosis involving the urinary bladder. No enlargement of the tumor or upper urinary tract obstruction has occurred during the long-term follow up. We recommend meticulous follow up of patients with giant intrapelvic neurofibromatosis. PMID- 11903696 TI - Post-traumatic high flow priapism: demonstrable findings of penile enhanced computed tomography. AB - Post-traumatic high flow priapism is a rare disease. A review of English published reports revealed 63 cases. Enhanced computed tomography (CT) of the penis has not previously been used as a diagnostic method for post-traumatic high flow priapism. We present a case of post-traumatic high flow priapism diagnosed with enhanced CT of the penis. Additionally, diagnostic modalities for post traumatic high flow priapism are discussed with review of published work. PMID- 11903697 TI - Report of a 45, X male with monoorchism and distal hypospadias. AB - We present the rare case of a 5-year-old boy with a 45, X karyotype. The boy's family pedigree analysis was unremarkable. On admission, he was 97.4 cm tall (2.0 SD below normal references) and weighed 13.9 kg (1.0 SD below normal references). Mild mental retardation was suspected on the PPVT scale. Physical examination revealed a well-developed penis with subglanular hypospadias. His left testis was located well down at the bottom of the scrotum and the right testis was impalpable. Unilateral testicular agenesis and persistence of mullerian remnants within the hernia sac were noted on the previous records of inguinal exploration on the right side. The left testis was biopsied through a scrotal incision and prepubertal testicular tissue was confirmed. No ovary was found on laparotomy exploration. Hypospadias was repaired with meatal advancement and glanuloplasty (MAGPI) urethroplasty. His postoperative course was uneventful. Our report represents a rare case of a boy with a 45, X karyotype. Current theories for testicular development were reviewed. PMID- 11903702 TI - Occupational therapy in accident and emergency departments: a randomized controlled trial. AB - BACKGROUND: The National Service Framework for Older People published in 2001 requires action to improve the discharge process of older people from hospital. Currently, many older people released from accident and emergency departments are unable to perform basic activities of daily living. This could delay recovery and increase demand for emergency primary care services. AIMS: To evaluate the potential for an occupational therapist in an accident and emergency department to reduce unmet functional needs after releasing patients aged 75 years or more with a primary diagnosis of limb, rib or back trauma. METHODS: A randomized controlled trial was employed, with the intervention group receiving an occupational therapy assessment and given, or arrangements made, for appropriate treatments and equipment before release. Controls received routine care. All patients were reassessed at home, 7 days later. RESULTS: Of 72 patients, 39 were recruited with 19 randomized to the intervention group and 20 to the control group. The median age was 81 years (75-92 years) and 31 (79%) were female. At baseline, 20 (51.2%) patients had problems in performing one or more of four basic activities assessed. At the follow-up assessment, in the intervention group the proportion of patients having no problems with these activities increased by 54% over and above the change in the control group, compared with that at baseline (P < 0.001). No effect on anxiety or demand for primary care was observed. CONCLUSIONS: In our local hospital, over 50% of older patients with limb, rib or back trauma would have left the accident and emergency department unable to perform basic activities of daily living. This might be overcome by employing occupational therapists to assess and meet the functional needs of these patients before sending them home. These results need confirming in a larger study. PMID- 11903701 TI - Oration. Nursing: a new era for action. PMID- 11903703 TI - You can do it if you set your mind to it: a qualitative study of patients with coronary artery disease. AB - AIMS OF THE STUDY: To gain increased knowledge and understanding of what it means to be afflicted with coronary artery disease (CAD) and how it affects the life/lifestyle of the individual. BACKGROUND: Research has documented that education, counselling and behavioural interventions are important elements of cardiac rehabilitation and compliance with treatment. Compliance is generally better with medical treatment than with recommended lifestyle changes. Another influencing aspect is locus of control, i.e. people's own understanding of control is the foundation for the decisions patients make more or less consciously regarding compliance with caring/nursing, treatment and lifestyle changes. METHODS: Eight individuals with diagnosed coronary artery disease were interviewed about their life situation, and the opportunities and obstacles they encountered in making lifestyle changes. These interviews were transcribed and then analysed using a hermeneutic approach. FINDINGS: The findings included three areas: (1) The causes of coronary artery disease describing different factors, such as heredity, lifestyle and demands. (2) Difficulties in the work of rehabilitation, which was explained in terms of informants' feelings of confusion, uncertainty and sadness. (3) Successful rehabilitation consisted of two factors: the personality of the individual patient and external support. CONCLUSIONS: Patients may comply well with follow-up visits but less with lifestyle changes. By identifying different 'characteristics', 'prerequisites' and 'difficulties' that describe patients' compliance, it should be possible to make treatment more individual. Nurses have a significant role in supporting these patients since they are more accessible than physicians. Nurses also have a responsibility to work together with patients to empower them, in order to make their lifestyle changes and self-care activities manageable. PMID- 11903704 TI - From hospital to home care: a randomized controlled trial of a Pain Education Programme for cancer patients with chronic pain. AB - AIM OF THE STUDY: To investigate the role of district nurses in the care of cancer patients with chronic pain at home, as well as the effects of a Pain Education Programme for patients and their district nurses. The Pain Education Programme consisted of a tailored multi-method approach in which they were educated about pain, instructed how to report pain, and how to contact health care providers. BACKGROUND: No educational programs for patients in pain have been studied in outpatients nor integrated with the home care provided. DESIGN AND METHODS: One hundred and four patients and their 115 district nurses were enrolled in a prospective, longitudinal, randomized controlled study. The primary outcome of interest was type of care provided by district nurses, satisfaction with the pain treatment, and agreement in estimating patients' pain intensity. RESULTS: Results showed that continuity of care was poor as only 36% of the district nurses were informed about patients' pain by hospital nurses. Pain was rarely the reason for referring the patient to district nursing after discharge. Although pain control was not a main reason for district nurses to visit a patient, pain was a subject for discussion in 76% of visits. Besides discussing the pain problem with patients, district nurses provided only a few pain relieving interventions. District nurses randomized to the intervention group significantly better estimated patients' pain intensity, and were more satisfied about patients' pain treatment, but no differences were found in their assessment of patients' pain relief. CONCLUSIONS: These findings suggest a significant but moderate effect of the Pain Education Programme, with district nurses only playing a minor role in the treatment of cancer pain. PMID- 11903705 TI - Mothers' evolving relationships with doctors and nurses during the chronic childhood illness trajectory. AB - AIMS: In this paper the evolution of mothers' relationships with doctors and nurses during the chronic childhood illness trajectory is explored and discussed and the implications for child health care are considered. The discussion reports one aspect of a qualitative study that sought to determine whether time of diagnosis influenced mothers' coping across the illness trajectory. BACKGROUND: The quality of relationships between mothers, who are usually the primary carers, and nursing and medical staff is central to the experience of coping with chronic childhood illness. Mothers need to develop expertise in a wide range of technical skills and knowledge of complex health care issues, while also coming to terms with the uncertainty of a condition with an unpredictable trajectory. Attempts to understand the significance of relationships between those living with chronic illness and staff have highlighted a number of factors that may influence the degree to which they are viewed as satisfactory. The way staff communicate with families has been identified as one of the mediating processes through which the quality of such is determined. DESIGN: Mothers of 15 children diagnosed presymptomatically and 14 children diagnosed postsymptomatically with the chronic illness Vesicoureteric Reflux (VUR) were interviewed to assess and compare coping strategies across the trajectory. Mothers' written consent was obtained and interviews were tape-recorded, transcribed and analysed using the 'Framework Technique'. FINDINGS AND CONCLUSION: The need to develop and sustain trusting relationships with staff was reported as a continual source of stress for mothers. The formation of satisfactory alliances, based on mutual respect and good communication early in the trajectory, in particular during the prediagnostic phase, was found to be instrumental in coping and competence development during the later chronic phase of the trajectory. Further research is needed to map prospectively the evolution of relationships between mothers and staff across the trajectory. PMID- 11903706 TI - 'Breaking bad news' within a paediatric setting: an evaluation report of a collaborative education workshop to support health professionals. AB - AIMS OF PROJECT: To evaluate a workshop to prepare health professionals for breaking bad news in the paediatric setting. BACKGROUND: Breaking bad news can be difficult for health professionals, and it seems that few receive specific training for undertaking this challenging task. Latterly, however, there have been reports of training programmes being developed to prepare health professionals for breaking bad news, although most focus upon meeting needs of medical staff only. While doctors have a uniquely important role in breaking bad news it is evident that other health professionals, most frequently nurses, can be equally involved in this encounter. Accordingly, nurses and other professionals need training to recognize the contribution that they can make in ensuring sensitive and effective bad news disclosure. DESIGN: A one-day, multi professional, experiential training workshop. METHOD: Forty-five participants, mainly nurses (34, 76%) and doctors (10, 22%), attended one of five breaking bad news workshops set up for staff working in a range of paediatric settings including Accident and Emergency and Intensive Care. Each of the workshops was facilitated by three facilitators from varied backgrounds. Using an experiential design, participants were supported to explore and reflect upon breaking bad news issues, which also included engagement with actors to act out realistic bad news scenarios. Debriefing, using a positive learner-centred model of feedback, provided the main platform for promoting learning. Following the workshop, participants completed an evaluation questionnaire, seeking their perceptions of the effectiveness of the workshop in enhancing knowledge and communication skills. DATA ANALYSIS: Atlas.ti, a qualitative computer data analysis software program was used to explore the evaluation comments made by participants, resulting in the generation of common themes. FINDINGS: Seven themes, including development of practice, the value of sharing, benefit of feedback, and team work, emerged from responses. All responses indicated that the workshop had been beneficial and an effective training method, with most participants (40 of 89%) indicating that they would strongly recommend their colleagues to attend a similar workshop. CONCLUSION: The educational approach reported is considered by participants to be beneficial in preparing health professionals for breaking bad news in a collaborative way. PMID- 11903707 TI - Development of quality of nursing care in Thailand. AB - BACKGROUND: In Thailand there is a need to establish definitions of quality of nursing care and to determine how it is measured. There is an urgent need to focus more closely on indicators of nursing quality. In Thailand there is no agreed upon definition of quality nursing care or indicators by which to measure it. PURPOSE: The purpose of this research was to develop a definition of quality of nursing care and to begin the process of defining quality of care indicator measures to evaluate the care given in acute care settings. METHODS: This descriptive study was conducted in Thailand. The process included two phases. The first phase used individual interviews and focus group discussion and the second phase included consultation with quality of nursing care experts. An interview guide was used to structure the interviews and the draft definitions of quality and suggested indicators were used when consulting with the experts in the second phase of the study. FINDINGS: The findings are organized into two sections: (1) definition of quality of nursing care and quality of nursing indicators; and (2) consultation with Thailand quality experts to refine the definition and quality of care indicators. The indicators were categorized into three groups: structure, process, and outcome. LIMITATIONS: This descriptive study using qualitative method relied upon the opinions of various stakeholders. Their opinions may or may not have any direct relationship to the scientific literature related to quality. Many of the indicators need to be further refined. CONCLUSIONS: The findings generally support the initial work done in the United States of America. There is a need to further refine the various Thailand indicators. This study is the first in Thailand that has attempted to address quality of nursing care. PMID- 11903708 TI - Profile of first-line nurse managers in New South Wales, Australia, in the 1990s. AB - AIMS: To determine a demographic profile, employment history and career plans of a sample of nursing unit managers (first-line managers) in New South Wales (NSW), Australia in 1999. To compare the profile of first-line nurse managers in 1999 with those in 1989. BACKGROUND: This study replicates another undertaken a decade earlier (see Duffield 1992). Tracking the changes to nurse manager positions is important, given changes to the educational preparation of nurses and restructuring within hospitals which have occurred in the past decade. METHOD: The same questionnaire was used in both studies, with minor amendments and the addition of two items to reflect changes to organizational structures, whereby nurse managers are now responsible for non-nursing staff. In 1999 all first-line nurse managers in the largest health service in NSW were invited to participate in the study. RESULTS: A response rate of 77% was achieved. There were few differences of note in the demographic profile from 1989 to 1999. However educationally, first-line nurse managers in 1999 were more highly qualified. A greater proportion had higher degrees and increasingly, in the management discipline. CONCLUSION: Perhaps reflecting these qualifications, more managers in 1999 indicated their intention to move to more senior management positions in the next decade. The mode for experience in this role of 1 year in both 1989 and 1999 reflects a worrying trend of high turnover and inexperience amongst this group of managers. While maternity relief might account for this result, further research needs to determine more precise reasons. The ad hoc bases on which expert clinicians (clinical nurse specialists) act as the manager in his/her absence need to be critically examined. Alternative strategies such as introducing a formal second- in-charge position may ensure more successful recruitment and retention of staff to these critical management positions. PMID- 11903709 TI - A study of work stress, patient handling activities and the risk of low back pain among nurses in Hong Kong. AB - RATIONALE: Low back pain (LBP) remains a common and costly problem among the nursing profession. Several studies have indicated that LBP is attributed to mentally straining or demanding work, fatigue or exhaustion or general work satisfaction. AIMS: This study aims to measure the magnitude of LBP among Hong Kong nurses and its association with the work-related psychological strain and patients handling activities. RESEARCH METHODS AND MEASURES: A cross-sectional study of Hong Kong hospital nurses was conducted. Three hundred and seventy-seven nurses were recruited from six district hospitals. They were registered nurses or enrolled nurses working full-time for at least 1 month in the current ward. One hundred and seventy-eight (47.2%) study subjects were randomly selected from two district hospitals and 199 (52.8%) study nurses made up the convenience sample. Possible bias from psychological distress, socio-demographics and lifestyle factors was controlled for. Data were collected by face-to-face interviews. The data included work factors (both psychological stress and patient handling activities related to work), demographics, psychological distress and lifestyle factors and the occurrence of LBP. RESULTS: Of the 377 nurses interviewed, 153 (40.6%) reported having LBP within the last 12 months. With symptoms of LBP as the outcome, risks were increased where nurses self-reported that they only occasionally or never enjoyed their work [adjusted odds ratio (OR) 2.07], where frequent manual repositioning of patients on the bed was required (adjusted OR 1.84) and where they were required to assist patients while walking (adjusted OR 2.11) after adjustment for other potential confounders. CONCLUSION: The results indicate that an association exists between work stress, manual lifting and LBP prevalence. The main route to prevention of LBP among nurses is likely to lie in improved ergonomics and psychological health in their work place. Good posture and correct transferring techniques in ward situations should be reinforced with hands-on practice performed on nurses' common types of clients. PMID- 11903713 TI - Families, nurses and intensive care patients: a review of the literature. AB - 1. Nurses striving to give holistic care to provide quality care for their patients, need to recognize the importance of caring for patients' families. 2. A detailed review of the literature examining the relationships between nurses and intensive care patients' families was undertaken to examine its strengths and weaknesses as a basis for further study. 3. Studies show that although nurses are often in the best position to meet families' needs, their needs are not always met. 4. The building of good relationships with families is essential for nurses, and yet evidence shows that some nurses have difficulties in this area. 5. Good practice is identified and obstacles nurses face in forming relationships with families are explored. 6. Strategies for improving the interaction process between intensive care nurses and patients' families are systematically evaluated. PMID- 11903710 TI - A stepwise multivariate analysis of factors that contribute to stress for mental health nurses working in the community. AB - AIMS: The aim of the study was to examine the variety, frequency and severity of stressors experienced by community mental health nurses (CMHNs) in Wales. BACKGROUND: Numerous studies undertaken throughout the United Kingdom (UK) have indicated that those health professionals working as part of community teams are experiencing increasing levels of stress and burnout. Sample sizes have tended to be small and participants have been drawn mainly from sites in England. METHODS: A questionnaire booklet, which included a number of validated measures, was distributed to 614 CMHNs. These included Maslach Human Services Survey, Community Psychiatric Nursing (CPN) Stress Questionnaire, PsychNurse Coping Questionnaire, Rosenberg Self-Esteem Scale and General Health Questionnaire GHQ-12. The study was the largest of its kind conducted in the UK. RESULTS: Data were collected from 301 CMHNs, representing a response rate of 49%. Community mental health nurses identified the most stressful issues as trying to maintain a good quality service in the midst of long waiting lists and poor resources and having too many interruptions while trying to work in the office. The best demographic predictors of high stress scores were having an unsupportive line manager, working with a specific client group and not having job security. These factors accounted for 20% of the variance in the total stress score. When the results from the psychometric instruments were included, 46% of the variance in the total stress score was accounted for. The predictive variables were emotional exhaustion, working with a specific client group, job security and alcohol consumption. CONCLUSIONS: These findings indicate that there is a need to create more supportive environments both in terms of job security and management support, especially for those working in the fields of severe mental illness and rehabilitation. PMID- 11903714 TI - Relocation stress in critical care: a review of the literature. AB - 1. Transfer to the ward following a period in intensive care may cause stress for patients. 2. A review of the literature reveals that this phenomenon has been described in a number of different ways, such as transfer stress, transfer anxiety, translocation syndrome and, more recently, relocation stress. 3. This paper reviews the various concepts before arriving at a more operational definition of the phenomenon. 4. It attempts to reveal what causes this phenomenon and to what extent it exists. 5. Patients' responses to transfer are identified and the physical and psychological problems that have been associated with discharge from intensive care are discussed. 6. Lists of interventions that the literature suggests may reduce or prevent this phenomenon from occurring are reviewed. 7. Recommendations for practice development and further research are made. PMID- 11903715 TI - Nursing activity in general intensive care. AB - 1. In this cost-conscious climate there is a need to make explicit and justify the rationale to support direct patient contact by Registered Nurses. The current shortage of qualified nursing staff means that it is essential that experience and expertise be utilized to the benefit of patients and the service as a whole. 2. This study used a descriptive approach to describe, categorize and quantify the activities of nurses working in a six-bed general intensive care unit. 3. Data were collected using a self-reporting diary log sheet that identified the focus of an individual's activity at 5-minute intervals. All Registered Nurses, on all shifts over a 7-day period, completed log sheets. 4. The results demonstrate that nurses working in this general intensive care unit spent 85% of their time in activities associated with providing direct patient care. However, up to 6% of time was spent undertaking non-nursing duties, and analysis of unit activity provided data to support an increase in the establishment and review of the shift patterns of health care assistants. 5. The findings of the study indicate that nurses in charge of shifts spend 24.1% of their time in managerial and administrative activity; this reduces the amount of time spent in direct patient contact. PMID- 11903716 TI - The hidden benefit: the supportive function of the nursing handover for qualified nurses caring for dying people in hospital. AB - 1. The nursing handover is a key activity for nurses working in acute hospital wards in the NHS. Little scholarly attention has been paid to the use nurses make of the information exchanged during nursing handover or how certain features of the nursing handover might impact positively or negatively on patient outcomes. 2. This paper draws on data from a phenomenological study of 28 qualified diplomate nurses. 3. During the course of non-directive semi-structured interviews, some of the participants in this study expressed the opinion that nursing handover was helpful in enabling them to work with dying people in the acute hospital medical wards in which they worked. 4. The nurses identified two important functions of the nursing handover. The first was as a forum for discussing opinions and expressing feelings. The second was as a source of information on which to base their nursing decisions and actions. 5. It is proposed that some qualified nurses need help with the emotional labour of caring for dying people and that the nursing handover can assist in emotional adaptation, so enabling the management of troubling thoughts or feelings experienced in the course of caring for someone who is dying. 6. The role of the nursing handover in providing emotional support for nurses has been little studied and is a potentially useful area of future research, especially if it can be related to patient experiences and outcomes. PMID- 11903717 TI - Mothers' experience of social support following the death of a child. AB - 1. This study aimed at analysing the grief and coping of mothers whose child had died under the age of 7 years. The paper describes the social support received as experienced by mothers. 2. Data were collected using a survey (n=91) and interviews (n=50) with mothers who had lost their child at least 1 year previously. The questionnaire contained questions concerning background characteristics, the Hogan Grief Reaction Checklist and open-ended questions. Survey data were analysed using a two-way analysis of variance, Wilcoxon test, cross-tabulation and content analysis. Interview data were analysed using inductive content analysis. 3. Findings showed that the spouse, children, grandparents, next of kin, friends and colleagues were the main sources of support. 4. Support consisted of emotional support, informational and instrumental support, and consolation and caring. Informational support consisted of advice and guidance from the mother's own mother or fellow sufferers. Instrumental support consisted of assistance with practical issues. Negative support manifested itself in unwarranted interference by relatives in the family's affairs or breaking up of friendships. 5. Mothers expected professional practitioners to provide honest information about the dying child's illness and practical arrangements after the child's death, and to keep up hope as long as the child was alive. 6. The care facility was also expected to maintain contact with the family after the child's death. PMID- 11903718 TI - Nurses' decision-making in collecting information for the assessment of patients' nursing problems. AB - 1. The paper addresses two questions: Firstly, what kind of information do nurses acquire from cancer patients for purposes of judging their patients' problems and preparing a care plan? Secondly, how systematically do nurses proceed in the decision-making process from the formulation of initial assumptions about the patient's situation to the final definition of problems? 2. The instrument used for data collection was a computer-simulated case description compiled by a team of four nursing researchers and one medical researcher. The case description was based on a real patient history. 3. The sample consisted of 107 Registered Nurses on four oncology, two internal medicine and five surgical wards of two central university hospitals in Finland. Data were collected in autumn 1998 and spring 1999 using a laptop computer and a tape recorder. 4. The four most important problems identified by nurses at baseline were pain (85%), pain medication (59%), family situation (66%) and spread of cancer (49%). Presented with a list of 23 options, they obtained additional information on average on 13 areas. Almost one third collected information from 16 to 22 areas. On average nurses identified 12 of the 28 nursing problems specified. A statistically significant association was observed between information acquisition and problem definition in seven different variables. These had to do with pain, general condition and prognosis. 5. Nurses adequately prioritized their patients' problems and systematically collected data on those problems. On the other hand they also identified a number of problems that were not relevant to the situation. PMID- 11903719 TI - Nurses' altered conceptions of work in a ward with all-RN staffing. AB - 1. The aim of this study was to investigate how nurses' conceptions of their patients and work changed after reorganization to all RN-staffing and the adoption of a patient-in-focus philosophy on the ward. 2. The study builds on the perspective that the individual's conception of work precedes and forms the basis for the development of knowledge, skills and attributes used in accomplishing work. 3. The findings are based on a secondary analysis of two open interviews with 22 nurses on the ward. These interviews were conducted on two occasions with an interval of 2 years. The third interview was carried out 6 months later, when 10 nurses were asked to talk about a patient's care episode in a narrative form. 4. The nurses' conceptions changed towards a holistic view of the patient, they developed a new approach to work and they used the altered circumstances in their work. PMID- 11903720 TI - Barriers to and facilitators of research utilization among Finnish registered nurses. AB - 1. The aim of nursing research is to produce a sound foundation for evidence based nursing; the job of nurses is to make the best possible use of that foundation and apply the knowledge produced to the practice of nursing. 2. The purpose of this study was to identify and describe barriers to and facilitators of research utilization from the point of view of Finnish Registered Nurses. 3. The BARRIERS Scale was administered to 316 nurses in two major hospitals; 253 nurses returned the questionnaire, giving a response rate of 80%. The structured data were processed with SPSS 9.0, and the unstructured data were interpreted using the method of content analysis. 4. The main barriers to research utilization identified by the respondents were: the fact that most research is published in a foreign language; that physicians will not co-operate with implementation; and that statistical analyses are difficult to understand. The facilitators mentioned most often were nurses' positive attitudes and abilities. Other important facilitators included the support and activity of a ward sister as well as encouragement, a favourable attitude and collaboration on the part of all staff members. 5. The findings are discussed in relation to the Finnish healthcare context and nurse education, and evidence-based nursing practice. PMID- 11903721 TI - Cognitive behavioural therapy for clients with schizophrenia: implications for mental health nursing practice. AB - 1. This paper discusses the application of cognitive behavioural therapy (CBT) in the treatment of clients with schizophrenia and the implications for mental health nursing practice. 2. The literature shows that CBT has positive effects for clients in improving mental state and reducing relapse rate. 3. Because of their direct client contact, mental health nurses are in the best position to assess, address and manage clients' psychotic symptoms and problems. 4. A nursing care plan is used to illustrate how the principles of CBT could be incorporated into nursing care. 5. Education and training are important to equip nurses with the necessary knowledge and skills to implement the therapy. 6. Nurses have to take into account cultural issues related to the use of CBT. 7. Culturally sensitive research needs to be conducted to assess the outcome of the therapy. PMID- 11903722 TI - Carers' interactions with patients suffering from severe dementia: a difficult balance to facilitate mutual togetherness. AB - 1. A phenomenological-hermeneutic approach was used to illuminate carers' video recorded interactions in connection with supervision for individualized nursing care. 2. In order to disclose any changes in the carers' interactions with patients suffering from severe dementia the video recordings were conducted before, during and after the intervention. 3. The content of the videos was transcribed as a text, mainly verbal communication. Due to the rich data the videos and text were kept together as a whole in every step of the analysis. 4. After an initial naive understanding, different subthemes emerged in the structural analyses: promoting competence, struggling for co-operation, deep communication for communion, showing respect for the unique person, skills in balancing power, distance in a negative point of view, and fragmentary nursing situations. 5. The overall theme was 'Carers' balancing in their interactions, verbal as well as non-verbal, to promote a sense of mutual togetherness with the patient'. 6. The supervision intervention contributed to an improvement in carers' skills in balancing in their interactions. In the caring process carers' and patients' shared experiences and, due to patients' disabilities, interactions depended mainly on carers' qualities and capabilities for this confirming nursing care. PMID- 11903723 TI - The relationship between organizational climate and the content of daily life for people with dementia living in a group-dwelling. AB - 1. One factor influencing the outcome of care may be nursing staff's experience of the organizational work climate. The aim of the study was to investigate how people with dementia spend their time in group-dwelling units (GD) with either a creative or less creative organizational climate. 2. For the study, two GD units assessed as having a creative organizational climate and two units assessed as having a less creative climate were selected. Eighteen residents living in the units assessed as creative and 20 residents living in the units assessed as less creative participated in the study. 3. For measuring the organizational climate the Creative Climate Questionnaire was used. Observations of residents' activities were classified according to the Patient Activity Classification. For measuring residents' functional ability the Multi-Dimensional Dementia Assessment Scale was used. Their cognitive capacity was measured with the Mini Mental State Examination. 4. Residents living in the units assessed as having a creative organizational climate spent 45.2% of the time with nursing staff, while those in the less creative climate spent 25.6% (P < 0.001). Time spent with fellow residents in the creative climate was 13.9% and in the less creative climate 31.3% (P < 0.001). There was no significant difference between the units according time spent with relatives and time spent alone. 5. Since the purpose of GD is to offer care adapted to the abilities and psychosocial needs of people suffering from dementia, a less creative climate can be a threat to the aims of GD. In order to maintain these, it is important for managers to be aware of the work climate and its impact on care for people with dementia. PMID- 11903724 TI - The meaning of fatigue and tiredness as narrated by women with fibromyalgia and healthy women. AB - 1. The aim of this study was to elucidate the meaning of fatigue and tiredness as narrated by women with fibromyalgia (FM) and healthy women. Twenty-five women with FM were interviewed with a narrative approach about the meaning of the lived experience of fatigue and tiredness. A reference group of 25 healthy women was interviewed about the same topic. 2. A phenomenological-hermeneutic method inspired by the French philosopher Ricoeur was used to interpret the interview text. 3. The meaning of fatigue and tiredness was related differently by women with FM and healthy women. The findings are presented in four major themes for women with FM -- the body as a burden, an absent presence, an interfering obstacle and being in hope of alleviation -- and in one major theme for healthy women: needing recovery. 4. Women with FM narrated fatigue as making it obvious that I have a body, instead of I am my body; the lived body becomes urgently present, as an 'it'. Healthy women narrated tiredness as a natural phenomenon when they need recovery and time to rest. 5. The findings are interpreted in the light of the phenomenological work on the lived body by Leder, Toombs and Merleau Ponty. PMID- 11903725 TI - A study of parenteral use of methotrexate in rheumatic conditions. AB - 1. This paper reports the findings of a small pragmatic study to compare the safety and efficacy of methotrexate administered by intramuscular and subcutaneous injection, and to teach patients to self-administer methotrexate by the subcutaneous route. 2. Eight patients with rheumatic conditions, already receiving a stable weekly dose of methotrexate by intramuscular injection, were entered into this 13-week study. 3. Serum levels of methotrexate were measured on six consecutive occasions: three whilst patients received intramuscular methotrexate and then three after switching to the subcutaneous route. 4. Patients were taught to self-administer their methotrexate subcutaneously and were then discharged to perform this task at home. 5. Levels of disease activity and psychological scores were measured at the start and end of the study. Satisfaction with self-administration and teaching of injection techniques were assessed at 13 weeks. 6. Serum methotrexate levels were not significantly affected by the route of administration. All patients were able to perform self injection safely and seven out of eight preferred self-administration at home. 7. This small study demonstrates that there is no difference in the safety and efficacy of methotrexate given by either parenteral route. Patients were able to administer safely methotrexate subcutaneously. Self-administration reduced hospital visits, was more convenient for patients and improved patient satisfaction. PMID- 11903727 TI - Working with women experiencing mid-trimester termination of pregnancy: the integration of nursing and feminist knowledge in the gynaecological setting. AB - 1. Working with women experiencing a mid-trimester termination of pregnancy is part of clinical practice in many gynaecological services. In this paper recent research with nurses working in the gynaecological area is drawn on to explore the issues for nurses working with women experiencing mid-trimester termination. Mid-trimester terminations are those carried out between approximately 12 and 20 weeks. 2. Mid-trimester termination results in the delivery of a fetus and this event requires sensitive management as it is has the potential to cause distress for the women due to the psychological and physical impact of the procedure. However, health professionals involved can also find this a distressing clinical event due to the complex nature of the management and care required. 3. Consideration of this clinical event from a feminist perspective led to my exploring the way in which feminist theory could be applied in the situation of mid-trimester termination. Using notions from feminist theory can assist in the management of this process, and feminist concepts related to the centrality of women's experience can be integrated into actual practice. 4. A series of recommendations are provided in this paper to show the way in which feminist concepts can be integrated into clinical practice. Integrating feminist principles into practice can support both the woman experiencing the abortion and the nurse whose role in the event is sustained and intimate, and result in positive outcomes for both women. This can result in an environment that is safe and supportive for all women involved in mid-trimester terminations. PMID- 11903726 TI - Investigating the effect of erectile dysfunction on the lives of men: a qualitative research study. AB - 1. The aim of this project was to identify and explore the issues facing men who live with erectile dysfunction (ED), in particular men's' relationships with women partners and men's interactions with the wider world. 2. In order to gain an understanding of their everyday lives, a qualitative research design was used. This is an account of the interpretation and analysis of nine interviews with men living with ED that were carried out during the autumn of 1997. 3. The analysis identified two main themes, 'loss' and 'being alone with it'; with meta categories 'making sense of it' and 'telling other people', and 'place of sex'. The latter acts as a bridge between the two themes. 4. The implications for nursing practice are considered and recommendations are made for practice, education and research. PMID- 11903729 TI - Use of a bereavement service among suddenly bereaved families in Hong Kong. PMID- 11903728 TI - Sleep quality and responses to insufficient sleep in women on different work shifts. AB - 1. The aim of this study was to examine degrees of cognitive behavioural effects of fatigue, mood changes and somatic responses to sleep loss in women with and without sufficient sleep, and to explore possible links between effects of sleep loss and specific sleep disturbances in selected groups. 2. A total 156 women working in a casualty department on different work shifts responded to a questionnaire which measured sleep quality, strain and symptoms related to working conditions, as well as effects of sleep loss. 3. About 40% of the women had perceived insufficient sleep during the last 6 months. They perceived significantly worse sleep quality and a higher degree of strain according to working conditions than the others. Palpitation and dysphoria as effects of sleep loss were independently predicted by sleep quality. Dysphoria was also predicted by difficulty in falling asleep. Cognitive behavioural effects of fatigue was predicted by disturbed sleep. Palpitation effects led to a 10-fold increase in the probability of cognitive behavioural effects of fatigue. The effects were most prominent among women suffering from gastrointestinal problems of long duration and chronic pain. 4. Responses to reduced sleep quality in women constitute a form of stress, with sympathetic activation, increased susceptibility to infection, moderate cognitive impairment, mood changes and somatic distress. PMID- 11903730 TI - HIV drug therapy: a nurse-led approach. PMID- 11903731 TI - Patterns of reflux in gastroesophageal reflux disease: are they of clinical use? PMID- 11903732 TI - Membranous obstruction of the inferior vena cava (obliterative hepatocavopathy, Okuda). AB - Confusion prevails throughout the world regarding the definition and classification of the Budd-Chiari syndrome. The original patients (Budd and Chiari) described had hepatic vein thrombosis, but this syndrome now encompasses various hepatic venous outflow blocks, of which membranous obstruction of the inferior vena cava (IVC) is the most common. This author has been suggesting that the classical Budd-Chiari syndrome or hepatic vein thrombosis and membranous obstruction of IVC or primary thrombosis of IVC at its hepatic portion are epidemiologically, pathologically and clinically different, and that they should be treated as two clinical entities that are not to be mixed. The two diseases have a different onset, different clinical manifestations and a different natural history. Whereas hepatic vein thrombosis is a severe disease with an acute onset, IVC thrombosis presents mildly at onset, but it recurs and eventually turns into a fibrous occlusion of IVC of varying thickness or stenosis of a various degree. The fibrous IVC occlusion is found as a mysterious thin membrane, but is more often much thicker than a membrane, and therefore 'membrane' is a misnomer. Although the genesis is not established, formation of a thin membrane may be an outcome of recurrent thrombosis. The past congenital vascular malformation theory no longer holds, because the disease occurs mostly in adulthood, and transformation of thrombosis into a membrane has now been well documented pathologically as well as clinically. This author suggests that a term 'obliterative hepatocavopathy' replace membranous obstruction of IVC, the term 'Budd-Chiari syndrome' be abandoned, and that primary hepatic venous outflow block be divided into primary hepatic vein thrombosis and primary IVC thrombosis (obliterative hepatocavopathy). PMID- 11903733 TI - Upright versus supine reflux in gastroesophageal reflux disease. AB - BACKGROUND: Postural measures are early recommendations in the management of heartburn, and are aimed at preventing acid reflux through an incompetent lower esophageal sphincter (LES). However, LES incompetence is found in only a minority of patients, and transient LES relaxations, primarily in the upright position, are currently recognized as the main pathophysiological abnormality in gastroesophageal reflux disease (GERD). We investigated the importance of supine acid reflux in patients with GERD. METHODS: Upon review of their clinical, manometric, pH monitoring and endoscopic characteristics, 85 patients with reflux symptoms were classified into three groups: Group A (n=22), consisting of symptomatic patients without esophagitis or pathological reflux; group B (n=38), symptomatic patients with reflux but no endoscopic esophagitis; and group C (n=25), symptomatic patients with both ulcerative or complicated esophagitis and pathological reflux. RESULTS: All groups were similar in age distribution. Groups B and C had a higher prevalence of hiatal hernia and reflux symptoms. Manometry revealed similar LES pressures in groups A and B, but lower LES pressure in group C (P < 0.005). In groups A and B, supine reflux, in terms of percentage of time with pH < 4, was less pronounced than upright reflux (P < 0.0001). In contrast, group C supine reflux was as pronounced as the upright reflux. CONCLUSIONS: The majority of patients reflux in the upright position. Only patients with complicated esophagitis have significant bipositional acid reflux. These findings suggest that unless the patient has severe reflux disease, postural measures may not be indicated. PMID- 11903734 TI - Predominant nocturnal acid reflux in patients with Los Angeles grade C and D reflux esophagitis. AB - BACKGROUND AND AIMS: Nocturnal gastric acid breakthrough (NAB) is defined as an intragastric pH < 4.0 lasting more than 1 h during the night in patients taking a proton pump inhibitor (PPI). Gastroesophageal reflux disease (GERD) patients with nocturnal gastroesophageal acid reflux accompanied by NAB are thought to be refractory to PPI treatment. The aim of this study was to endoscopically identify the patients with predominant nocturnal gastroesophageal acid reflux. METHODS: The subjects were 37 patients with erosive reflux esophagitis (Los Angeles classification (LA) grade A, 12; B, 10; C, eight; and D, seven cases) and a control group of 20 patients without esophagitis. The results of ambulatory 24 h gastric and esophageal pH monitoring were compared among different grades of esophagitis. RESULTS: Gastroesophageal reflux during 24 h in patients with high grade esophagitis was more frequent than for patients with low-grade esophagitis or no esophagitis. Although the length of esophageal acid exposure (percentage time with pH < 4.0) in patients with grade A or without esophagitis was longer in the daytime, that in patients with grades C and D was longer during the night. The reason for the delayed nocturnal acid exposure was the longer nocturnal acid clearance in high-grade reflux esophagitis. CONCLUSIONS: Nocturnal exposure of the esophagus to acid occurs frequently in patients with LA grades C and D esophagitis. Thus, the existence of NAB with resulting nocturnal acid reflux should be considered when the patient with high-grade esophagitis shows resistance to PPI treatment. PMID- 11903735 TI - Helicobacter pylori alters n-6 fatty acid metabolism and prostaglandin E2 synthesis in rat gastric mucosal cells. AB - BACKGROUND AND AIMS: Little is known about whether Helicobacter pylori infection alters fatty acid metabolism in gastric mucosal cells. By using cultured rat gastric mucosal cells (RGM-1), we investigated the effect of H. pylori broth culture filtrates on this point. Furthermore, our study aimed to find out whether n-6 long chain polyunsaturated fatty acids from linoleic acid are formed in RGM-1 cells. METHODS: Rat gastric mucosal cells were incubated with 10, 20 and 40 microg/mL of linoleic acid or medium alone. Phosphatidylcholine content extracted from whole RGM-1 cells was quantitated by using a densitometer, and its fatty acid composition was analyzed by using gas chromatography. Prostaglandin E2 concentration in the culture medium was measured by using radioimmunoassay. The expression of cyclooxygenase (COX)-1 and COX-2 was examined by using reverse transcription-polymerase chain reaction. In addition, after incubation with [1 14C] linoleic acid, radioactivities of both linoleic acid and arachidonic acid components of the PC fraction were counted. The effects of H. pylori broth culture filtrates on PC content, its fatty acid composition and prostaglandin (PG)E2 synthesis were also assessed. RESULTS: Linoleic acid addition caused an increase in the composition of arachidonic acid, as well as linoleic acid, and also in PGE2 concentration. Cyclo-oxygenase-2 expression was induced in RGM-1 cells by the addition of linoleic acid. In addition, [1-14C] linoleic acid added to the culture medium was converted to [1-14C] arachidonic acid in RGM-1 cells. Helicobacter pylori broth culture filtrates decreased linoleic acid composition and increased arachidonic acid composition. Moreover, after incubation with H. pylori broth culture filtrates, PGE2 concentrations were higher than that of the controls. CONCLUSIONS: These findings suggest the presence of fatty acid elongase and Delta5- and Delta6-desaturases synthesize arachidonic acid from linoleic acid in RGM-1 cells. Thus, H. pylori infection may enhance PGE2 synthesis and accelerate n-6 fatty acid metabolism in gastric mucosal cells, which could make the gastric mucosal barrier more fragile. PMID- 11903736 TI - Vitamin C inhibits corpus gastritis in Helicobacter pylori-infected patients during acid-suppressive therapy. AB - BACKGROUND: Previous studies have shown that gastric acid suppression worsens corpus gastritis in Helicobacter pylori (H. pylori)-positive patients. We evaluated the effect of acid-suppressive therapy and vitamin C on H. pylori associated gastritis. METHODS: Forty patients with reflux esophagitis were divided into three groups by the status of H. pylori and therapy: group A (n=15), H. pylori (+) and omeprazole 20 mg; group B (n=15), H. pylori (+) and omeprazole 20 mg + vitamin C 1200 mg; and group C (n=10), H. pylori (-) and omeprazole 20 mg. In all three groups, the mucosal interleukin (IL)-8 contents, H. pylori colonization density, neutrophil infiltration in the corpus, and serum gastrin were evaluated at entry and 2 weeks after starting therapy; in group B, serum vitamin C levels were also measured. RESULTS: In group A, the IL-8 contents and the degree of neutrophil infiltration during therapy exceeded those at entry, whereas in groups B and C, these values did not change significantly with treatment. Helicobacter pylori colonization density during therapy was similar to that at entry in all three groups. The serum gastrin (in all groups) and vitamin C levels (in group B) during therapy exceeded those at entry. CONCLUSIONS: Potent acid suppression worsens H. pylori-associated corpus gastritis, although such worsening gastritis may be inhibited by vitamin C. PMID- 11903737 TI - Influence of Helicobacter pylori infection and cetraxate on gastric mucosal blood flow during healing of endoscopic mucosal resection-induced ulcers. AB - BACKGROUND AND AIM: Helicobacter pylori (H. pylori) infection is known to affect the gastric microcirculation, and cetraxate is reported to accelerate gastric ulcer healing, possibly by augmenting gastric mucosal blood flow (MBF). The aim of this study is to clarify the effect of H. pylori infection and cetraxate on MBF during gastric ulcer healing. METHODS: Forty-two patients who had undergone endoscopic mucosal resection (EMR) were studied. Mucosal blood flow was measured by the use of a laser Doppler flowmeter in the surrounding mucosa and at the ulcer margin, before, 1 day, 1 week and 4 weeks after EMR. Helicobacter pylori infection was confirmed by the use of bacterial culture and histology. After EMR, patients were randomly assigned to receive 30 mg lansoprazole (u.i.d; L-regimen) or 30 mg lansoprazole (u.i.d.) with 200 mg cetraxate (q.i.d; LC-regimen) for 4 weeks. RESULTS: The MBF ratio (MBF at ulcer margin/MBF in surrounding mucosa) 1 week after EMR was significantly lower than that before or 4 weeks after EMR only in H. pylori-positive patients treated with the L-regimen. No such decrease in MBF was observed after 1 week in H. pylori-positive patients treated with the LC regimen or in H. pylori-negative patients. CONCLUSION: A transient decrease in MBF was detected at the ulcer margin during healing of EMR-induced ulcers in H. pylori-infected patients. Cetraxate seemed to prevent this decrease in MBF. PMID- 11903738 TI - Influence of Helicobacter pylori infection on the prevalence of reflux esophagitis in Japanese patients. AB - BACKGROUND AND AIM: Reflux esophagitis is caused by esophageal motor dysfunction in patients with sufficient gastric acid secretion. Helicobacter pylori causes atrophic gastritis and influences gastric acid secretion. Hiatus hernia (HH) of the esophagus causes motor dysfunction in the lower esophagus. Therefore, this study aimed to test whether H. pylori infection, gastric mucosal atrophy and HH are predictive factors for reflux esophagitis. METHODS: Helicobacter pylori infection was examined in 781 patients by the measurement of serum immunoglobulin (Ig)G antibody, bacteriological culture and histological examination of biopsy specimens. The prevalence of HH, endoscopically identified gastric mucosal atrophy (closed- or open-type) and reflux esophagitis were investigated by reviewing endoscopic films. Investigated patients were divided into three age groups, under 49, 50-69, and over 70 years. The prevalence of esophagitis, H. pylori infection, gastric mucosal atrophy, and HH were compared to identify the possible predictive factors for reflux esophagitis by using logistic regression analysis. RESULTS: Sixty-nine patients with reflux esophagitis were found among the 781 investigated cases. The odds ratios of negative H. pylori infection, endoscopically identified closed-type gastric mucosal atrophy, and HH for the prevalence of reflux esophagitis were 1.342, 1.751 and 5.527, respectively. These results indicated that the presence of H. pylori infection was only a weak negative risk factor, and that HH was the most reliable endoscopic predictive factor for reflux esophagitis. CONCLUSION: Helicobacter pylori infection is a weak negative risk factor for the prevalence of reflux esophagitis, while HH is the most reliable predictive factor. PMID- 11903740 TI - Expression of B7 costimulatory molecules by cells infiltrating the colon in experimental colitis induced by oral dextran sulfate sodium in the mouse. AB - BACKGROUND AND AIM: T-cell activation, mediated by the interaction with major histocompatibility complex (MHC)-peptide complexes and B7 costimulatory molecules on antigen-presenting cells, is an essential event in the pathogenesis of inflammatory bowel disease (IBD). We investigated the expression of B7 costimulatory molecules on cells in the colon in an experimental mouse model of IBD to determine whether the B7/ligand interaction could provide a target for therapeutic intervention in IBD. METHODS: Experimental colitis was induced in mice by oral consumption of water substituted with 5% dextran sulfate sodium (DSS). Mice (n=4) were killed 1, 2, 3, 4 and 7 days after commencing DSS consumption, and colonic tissue was collected and examined immunohistochemically for T cells, B cells, macrophages and cells expressing B7-1 or B7-2. RESULTS: Compared to control mice drinking water, macrophage numbers in the colonic epithelium were elevated sevenfold by day 1 and T cells were elevated threefold by day 3 following commencement of DSS consumption. Numbers of infiltrating B7 positive (B7+) cells were not significantly elevated until day 7 when B7-1+, B7 2+ cells and macrophages were increased 20-fold compared to normal mice. CONCLUSION: These results demonstrate that an initial and rapid infiltration of the colonic epithelium by B7-negative macrophages is followed by an infiltration of T cells and subsequent upregulation of the B7 costimulatory molecules potentiating the inflammatory reaction in this disease model. These results suggest an intervention strategy based on the blockade of the B7-costimulatory axis could find application in the treatment of inflammatory bowel disease. PMID- 11903739 TI - Effect of omeprazole-induced achlorhydria on trefoil peptide expression in the rat stomach. AB - BACKGROUND: Omeprazole is an inhibitor of the H+K+ ATPase of the gastric parietal cell, which is used clinically to suppress gastric acid secretion. It has also been found to inhibit gastric mucin production; however, its effects on the synthesis and secretion of the trefoil peptides, which are also expressed by mucus cells, and which play a key role in cytoprotection and epithelial repair, are unknown. METHODS: Rats (n=8) were given either omeprazole (30 mg/kg per day; p.o.) or inert carrier for 1 week, and the effects on synthesis and peptide expression of the gastric trefoil peptides, TFF1/pS2 and TFF2/SP, were compared. RESULTS: As expected, omeprazole treatment abolished H+ ion production with a mean gastric juice pH of 7.2 compared with 2.4 for controls. The omeprazole group had elevated total protein levels of 35-fold and TFF1/pS2 peptide levels elevated fourfold, respectively, but not TFF2/SP peptide in gastric juice, suggesting that the increased pH reduced the viscosity of adherent mucus, thereby increasing gastric juice concentrations by dissolution of adherent TFF1/pS2 and increased secretion. Concomitant with increased TFF1/pS2 secretion was a fall in predominantly antral mucosal trefoil peptide concentrations. In contrast to trefoil secretory rates, the steady-state synthesis of both TFF1/pS2 and TFF2/SP was unchanged after omeprazole treatment, implying both a large cellular pool of processed peptide and rapid secretion. CONCLUSION: The increase in the concentration of TFF1/pS2 in gastric secretions during chronic omeprazole-induced achlorhydria may be important in preventing tissue injury and promoting repair in response to an increased luminal bacterial population. PMID- 11903741 TI - Chronic narcotic use in inflammatory bowel disease patients: prevalence and clinical characteristics. AB - BACKGROUND: Narcotic addiction can be a significant problem in inflammatory bowel disease (IBD). However, there are few published reports about this problem. METHODS: All patients prescribed narcotics chronically in the absence of demonstrable organic pathology were identified on the computerized Brisbane IBD Research Group database (n=332 patients with informative data as of 1 January 1999). Individual case records were reviewed with regard to clinical, psychiatric and social characteristics of these patients, and the prevalence of psychiatric disorders were compared with a control group of IBD patients. RESULTS: Eleven patients were identified. Nine had complete datasets, eight with Crohn's disease (CD), of which six had previous stricturing ileal disease, and one patient had ulcerative colitis, making a prevalence of 2.7% of IBD patients and 5.1% of CD patients. A 67% prevalence of a psychiatric disorder in narcotic users was significantly greater than the 8% prevalence in the control group of IBD patients (odds ratio 22, 95% CI 3.24-177). CONCLUSIONS: A significant proportion of IBD patients without demonstrable organic pathology were chronic narcotic users. Psychiatric disorders are common in this subgroup, as with chronic functional abdominal pain syndromes. It is suggested that inappropriate narcotic use in IBD patients can be reduced by appreciating that narcotics are a temporary therapy only for IBD patients, and awareness of pre-existing social and psychiatric disorders, which not only impact on clinical presentation of pain, but also help define the subgroup of patients who are at risk of narcotic misuse. PMID- 11903742 TI - Glucose-6-phosphate dehydrogenase deficiency is associated with increased initial clinical severity of acute viral hepatitis A. AB - BACKGROUND AND AIM: In glucose-6-phosphate dehydrogenase (G6PD) deficiency, the enzyme is deficient in liver cells as well as in erythrocytes. It has been suggested that this may be associated with a more severe clinical presentation of acute viral hepatitis A. The aim of this study is to determine the severity of liver disease in patients with viral hepatitis and G6PD deficiency. METHODS: Eighteen patients with diagnosed G6PD deficiency and acute hepatitis A were compared with 18 matched control patients with hepatitis A in a university hospital for liver disease severity and clinical outcome. RESULTS: Two of 18 patients with G6PD deficiency had neurological deterioration. Patients with G6PD deficiency had a mean peak prothrombin time (PT) that was significantly prolonged as compared with the control group (15.5 +/- 3.7 vs 12.9 +/- 2.0 s, respectively, P < 0.02), and a significantly higher proportion had an abnormal PT (PT > 13.3 s): 61 versus 11% (P < 0.0001). Hemolysis occurred in 44% of the G6PD deficiency patients. Total and direct bilirubin were significantly higher in all patients with G6PD deficiency, including patients without hemolysis. There was no significant difference in liver enzyme levels between the two groups. Patients with G6PD deficiency had a longer average hospital stay (9.5 +/- 4.8 vs 3.4 +/- 0.8 days, respectively, P < 0.001). There was no difference in the final clinical outcome between the two groups, and recovery of liver function was seen in all patients. CONCLUSIONS: Glucose-6-phosphate dehydrogenase deficiency in patients with hepatitis A causes a more severe initial clinical presentation, but does not alter the final clinical outcome. PMID- 11903743 TI - Anti-oxidant ebselen causes the resolution of experimentally induced hepatic fibrosis in rats. AB - BACKGROUND: Hepatic fibrosis occurs because of injury to the liver parenchyma and biliary system. We have investigated the effect of an organic selenium anti oxidant, ebselen, in the resolution of experimentally induced hepatic fibrosis, and evaluated its effect on various paradigms involved in hepatic fibrosis. METHODS: Following pretreatment with phenobarbitone, liver fibrosis was induced in male Fischer 344 rats by using carbon tetrachloride treatment for 10 weeks. Carbon tetrachloride-treated rats were randomly assigned into two groups: (i) no ebselen; and (ii) ebselen administered for 3 weeks following a 10-week carbon tetrachloride treatment period. Normal controls were: (i) neither carbon tetrachloride nor ebselen treated; or (ii) ebselen treated for 13 weeks. Liver sections were stained with hematoxylin and eosin, Masson trichrome and stained for reticulin by using silver impregnation. Reverse transcription-polymerase chain reaction was used to analyze the steady-state levels of gene(s) involved in: (i) hepatic fibrosis, namely, transforming growth factor-beta1, procollagen I and III, tissue inhibitor of metalloproteinase-1 and matrix metalloproteinase-13; (ii) oxidative stress, namely, cytochrome P4502E1; and (iii) preneoplastic liver foci, namely, the placental form of glutathione-S-transferase. RESULTS: Histological staining showed that ebselen resolves carbon tetrachloride-induced hepatic fibrosis. Treatment with ebselen reduced steady-state levels of transforming growth factor-beta1, procollagen I and III, tissue inhibitor of metalloproteinase-1, cytochrome P4502E1 and placental form glutathione-S transferase transcripts, and increased transcripts of matrix metalloproteinase 13. CONCLUSION: These findings provide evidence that ebselen significantly causes the resolution of carbon tetrachloride-induced hepatic fibrosis in rats. PMID- 11903744 TI - Effect of alcohol withdrawal on liver transaminase levels and markers of liver fibrosis. AB - BACKGROUND AND AIM: Acute alcohol withdrawal causes changes in hepatic blood flow and metabolism that may result in liver damage. This study aims to assess liver function tests and markers of hepatic fibrogenesis following alcohol withdrawal in alcoholics with clinically compensated liver disease. METHODS: Serial liver function tests and clinical assessments were performed on 22 male alcoholics during alcohol withdrawal. Plasma tissue inhibitor of metalloproteinase 1 (TIMP1), an inhibitor of collagen degradation, and plasma amino-terminal procollagen III peptide (PIIINP), a collagen precursor molecule, were measured in these alcoholics and in 11 control subjects. RESULTS: Transaminase levels did not change significantly over 7 days when all subjects were analyzed together. However, 32% of subjects showed a marked transaminase rise. These subjects did not differ from the others in baseline characteristics or short-term outcome, but had a greater benzodiazepine requirement. Only one subject consumed paracetamol (acetaminophen; 1-2 g/day). He had the largest transaminase rise. By comparing PIIINP assays, intact PIIINP concentration appears to increase following alcohol withdrawal. The TIMP1 levels were elevated in alcoholic subjects, but did not change following withdrawal. CONCLUSIONS: Increasing PIIINP suggests that hepatic fibrogenesis increases, or hepatic clearance falls, during acute alcohol withdrawal. The TIMP1 elevation in these alcoholics suggests that the inhibition of collagen degradation occurs while liver disease is still compensated. The period following alcohol withdrawal may be a time of marked increased susceptibility to paracetamol. The biochemical changes we observed were not associated with adverse short-term outcome, but the cumulative effect after repeated episodes of abrupt withdrawal may be of concern. PMID- 11903745 TI - A Japanese herbal medicine, Sho-saiko-to, prevents gut ischemia/reperfusion induced hepatic microvascular dysfunction in rats. AB - BACKGROUND AND AIM: We have reported that gut ischemia/reperfusion (I/R) causes hepatic microvascular dysfunction. Nitric oxide (NO) has been found to be a modulator of the adhesive interactions between leukocytes, platelets, and endothelial cells. Sho-saiko-to (TJ-9), a Japanese herbal medicine, is reported to have protective effects against liver injury and to regulate NO production. The objective of this study was to determine whether TJ-9 affects hepatic microvascular dysfunction elicited by gut I/R, and to investigate the role of NO. METHODS: Male Wistar rats were exposed to 30 min of gut ischemia followed by 60 min of reperfusion. Intravital microscopy was used to monitor leukocyte recruitment and the number of non-perfused sinusoids (NPS). Plasma tumor necrosis factor (TNF)-alpha and alanine aminotransferase (ALT) activities were measured. In another set of experiments, TJ-9 (1 g/kg per day intragastrically) was administered to rats for 7 days. In some experiments, dexamethasone (ST) (2 mg/kg per day intravenously) was administered. RESULTS: In control rats, gut I/R elicited increases in the number of stationary leukocytes, NPS, and plasma TNF alpha and ALT activities, and these changes were mitigated by the pretreatment with TJ-9. Pretreatment with an NO synthase inhibitor diminished the protective effects of TJ-9 on the increase in leukostasis in the pericentral region, NPS, and plasma TNF-alpha levels, but not its effect on the increase in leukostasis in the midzonal region, total number of stationary leukocytes, or plasma ALT activities. Pretreatment with TJ-9 increased plasma nitrite/nitrate levels. The responses caused by gut I/R were attenuated by the pretreatment with ST. Pretreatment with an NO synthase inhibitor did not affect the effect of ST. CONCLUSIONS: These results suggest that TJ-9 attenuates the gut I/R-induced hepatic microvascular dysfunction and inflammatory responses such as TNF-alpha production in the early phase via enhancement of NO production, and sequential hepatocellular damage via its anti-inflammatory effect like corticosteroid effect. PMID- 11903746 TI - Effect of endotoxin pretreatment on hepatic stellate cell response to ethanol and acetaldehyde. AB - BACKGROUND AND AIM: The role of endotoxin in alcohol-induced liver damage is well recognized. How pre-exposure to endotoxin might affect alcohol injury is not known. We herein studied the effect of endotoxin pretreatment on hepatic stellate cell (HSC) response to ethanol and acetaldehyde. METHODS: Rat HSC (CFSC-2G) were exposed to media supplemented with 1 microg/mL lipopolysaccharide (LPS). This was followed by a 24 h exposure to media containing LPS plus 50 mmol/L ethanol or 175 micromol/L acetaldehyde. Lipid peroxidation, collagen, and tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6 and transforming growth factor (TGF) beta1 secretion were determined at the end of both periods of exposure. RESULTS: Lipopolysaccharide pretreatment did not modify lipid peroxidation induced by ethanol or acetaldehyde alone. Glutathione (GSH) content decreased to 4.2 +/- 0.5 and 16.3 +/- 0.8 nmol protein after exposure to ethanol or acetaldehyde alone, and decreased further with LPS pretreatment (2.4 +/- 0.2 and 2.7 +/- 0.3 nmol/mg protein, respectively). Oxidized GSH (GSSG) content increased in ethanol and acetaldehyde LPS-pretreated cells only. Collagen secretion increased to 988 +/- 82 and 1169 +/- 91 microg/10(6) cells after exposure to acetaldehyde or LPS alone. Lipopolysaccharide pretreatment enhanced collagen secretion significantly in both ethanol- and acetaldehyde-treated cells (969 +/- 56 and 1360 +/- 72 microg/10(6) cells, respectively). Interleukin-6 production increased to 288 +/- 48, 1195 +/- 86 and 247 +/- 35 pg/mL per 10(6) cells after ethanol, acetaldehyde and LPS exposure, and increased further with LPS pretreatment in ethanol-exposed cells (680 +/- 23 pg/mL 10(6) cells). CONCLUSION: Lipopolysaccharide pretreatment of HSC adds to the damage produced by ethanol and acetaldehyde by diminishing GSH content and increasing GSSG content, collagen and IL-6 secretion. PMID- 11903747 TI - High viral loads, serum alanine aminotransferase and gender are predictive factors for the development of hepatocellular carcinoma from viral compensated liver cirrhosis. AB - BACKGROUND AND AIMS: The aims of the present study were to determine the occurrence rate of hepatocellular carcinoma (HCC) and to assess the risk factors for the development of HCC in compensated viral liver cirrhosis. METHODS: Two hundred and thirty-nine cirrhotic patients (65 hepatitis B surface antigen (HBsAg) positive, 165 hepatitis C virus (HCV) antibody positive (anti-HCV), and nine with both HBsAg and anti-HCV positivity) were studied. The Kaplan-Meier method evaluated by a log-rank test was used to estimate the cumulative probability of HCC development. Independent predictors of HCC development were estimated by using the Cox proportional hazard regression analysis. RESULTS: Dual infection manifested as HBsAg and anti-HCV positive was the highest risk of HCC. Multivariate analysis indicated that anti-HCV positive, HBsAg positive, and lactate dehydrogenase were independent predictors of the development of HCC among individuals with viral cirrhosis. In the HBsAg-positive group, a high-titer of HBV-DNA (more than 3.7 log genome equivalents (LGE)/mL) was most predictive of HCC development. In the anti-HCV-positive group, male gender and a high-titer of HCV-RNA (more than 1.0 Meq/mL) were predictive factors for the development of HCC. CONCLUSIONS: Individuals with high viral loads should be monitored for the development of HCC. Clinical efforts at eradicating or reducing the viral load may reduce the risk for HCC. PMID- 11903748 TI - N-Acetylglucosaminyltransferase V as a possible aid for the evaluation of tumor invasiveness in patients with hepatocellular carcinoma. AB - BACKGROUND: A close relationship has been shown to exist between the metastatic potential and beta1-6 branched oligosaccharides in human and rodent cells. N acetylglucosaminyltransferase V (GnT-V) catalyzes this process. Although this phenomenon has been reported, little is known about the clinical usefulness of the determination of GnT-V in the evaluations of tumor invasiveness in hepatocellular carcinoma (HCC). In this study, we measured the GnT-V activity in serum of patients with HCC, together with its activity and gene expression in HCC tissues, and elucidated the clinical usefulness of the GnT-V level in evaluating tumor invasiveness. METHODS: Seventy-three serum samples from 38 patients with HCC, 11 with chronic hepatitis, eight with hepatic cirrhosis and 16 healthy controls were used. Twenty-one liver tissues were obtained by surgical resection from 17 patients with HCC, three with colorectal cancers and one with gallbladder cancer metastatic to the liver. The GnT-V activity was determined by using high performance liquid chromatography. The GnT-V mRNA was quantified by using competitive RT-PCR. RESULTS: There were statistically significant correlations between GnT-V activity in sera of HCC, and GnT-V activity and GnT-V mRNA expression in tumor tissue. The mean GnT-V activity in the sera of patients with HCC increased in accordance with the degree of tumor invasion. The HCC group with intrahepatic and extrahepatic metastases showed the highest serum GnT-V-value. CONCLUSIONS: The present study demonstrated that there was a close association between tumor invasiveness and GnT-V activity in sera, and that the measurement of GnT-V may improve prognostic estimates and therapeutic outcomes for patients with HCC. PMID- 11903750 TI - Long-term outcome of autoimmune hepatitis in children. AB - BACKGROUND AND AIM: Autoimmune hepatitis (AIH) is a chronic disease of unknown etiology, which usually progresses to cirrhosis if not diagnosed and treated promptly. Data on long-term follow up in children with AIH are scant. The aim of this study is to assess the long-term outcome of autoimmune hepatitis in children with respect to clinical and laboratory features at presentation. METHODS: Data were extracted from the medical records of patients presenting over a 28-year period (1972-2000) to the Royal Children's Hospital, Melbourne, Australia. Additional information was obtained by interviewing patients, and their current physicians. Of the 30 patients (22 females, mean age 9 years) identified, 18 had type I, three had type II, four had autoimmune-polyendocrinopathy syndrome type 1, one had infantile giant-cell hepatitis associated with Coomb's-positive hemolytic anemia, and four were seronegative (antinuclear antibody (ANA), smooth muscle antibody (SMA) and liver-kidney microsomal antibody (LKM)). RESULTS: Clinical features at presentation included hepatomegaly (86%), jaundice (66%) and splenomegaly (50%). Initial investigations revealed a median serum bilirubin level of 55 micromol/L (range 6-425), median aspartate aminotransferase level of 678 IU (range 70-2548), and abnormal clotting in 33% of patients. Liver biopsies were performed on all patients at presentation and 11 showed cirrhosis (36%). The mean follow-up period was 10.0 +/- 7.8 years with 43% being followed for > 10 years. Only two patients died and one required transplantation. Fourteen (50%) patients continue to be on low dose prednisolone with azathioprine, two (7%) are on prednisolone alone, and six (21%) are on no therapy. When the cirrhotic and non-cirrhotic patients were compared, the albumin level at presentation was significantly lower in the cirrhotic group (P=0.01). Of the patients who were cirrhotic at presentation, six (54%) remain compensated with a mean follow-up period of 8 years. All 24 patients currently under follow up are engaged in age appropriate activities including school, part- or full-time work. CONCLUSION: Autoimmune hepatitis has a favorable long-term outcome with a transplant-free survival rate of 90% over a mean period of 10.0 +/- 7.8 years (range: 0.5-23), and a normal or near-normal lifestyle irrespective of presenting clinical, laboratory or histological features. PMID- 11903749 TI - Comparison of clinicopathological features of patients with hepatocellular carcinoma seropositive for alpha-fetoprotein alone and those seropositive for des gamma-carboxy prothrombin alone. AB - BACKGROUND: There has been no comparative study of the clinicopathological features of HCC patients who are seropositive for alpha-fetoprotein (AFP) alone and those who are seropositive for des-gamma-carboxy prothrombin (DCP) alone. The authors, thus, performed this comparative study. METHODS: The clinicopathological features of patients with solitary hepatocellular carcinoma (HCC), who underwent a hepatectomy were compared among the four below groups according to the seropositivity of AFP and DCP: group A, seronegative for both AFP below 20 ng/mL and DCP below 40 mAU/mL; group B, seropositive for AFP above 100 ng/mL and seronegative for DCP; group C, seronegative for AFP and seropositive for DCP above 100 mAU/mL; and group D, seropositive for both AFP and DCP. RESULTS: Group B patients showed a higher incidence of HCC with an indistinct margin, and a somewhat higher incidence of small HCC less than 2 cm in greatest dimension compared with group C patients. By contrast, group C patients had a higher frequency of HCC with a distinct margin compared with that of an indistinct margin, large tumors more than 3 cm compared with that of small tumors less than 2 cm, and a somewhat higher frequency of moderately to poorly differentiated HCC compared with that of well-differentiated HCC. Our HCC cases showed advanced clinicopathological features in the order of group C, group B and group A. Groups C and D patients showed similar characteristics. CONCLUSIONS: Hepatocellular carcinoma patients who were seropositive for AFP alone demonstrated clinicopathological features of less advanced HCC compared with those who were seropositive for DCP alone. PMID- 11903751 TI - Hepatobiliary and pancreatic: awoman with a back pain and increased urinary amylase. PMID- 11903752 TI - Gastrointestinal: diaphragmatic hernia. PMID- 11903753 TI - Therapeutic effects on intestinal Behcet's disease of an intravenous drug delivery system using dexamethasone incorporated in lipid emulsion. AB - Recurrent intestinal ulcer is a frequent problem in the management of Behcet's disease. However, no standard therapy for intestinal Behect's disease has been established. We report two patients with intestinal Behcet disease and recurrent ileal ulcers who were treated successfuly with a lipid emulsion of dexamethasone. In one patient, the cecal ulcer did not relapse after the intravenous administration initiation of a lipid emulsion of dexamethasone once a week, despite the discontinuation of prednisolone. In the other patient, the cecal ulcer showed a healing tendency, and oral administration of prednisolone was reduced from 40 to 15 mg/day after intravenous administration of a lipid emulsion of dexamethasone. Both patients experienced no complications associated with the administration of the emulsion. These results suggest that an intravenous drug delivery system using a lipid emulsion of dexamethasone is useful for treatment of intestinal Behcet's disease. PMID- 11903755 TI - Occult hepatocellular carcinoma with high fucosylated alpha-fetoprotein. PMID- 11903757 TI - Self-restriction of medications due to cost in seniors without prescription coverage. AB - OBJECTIVE: Little is known about patients who skip doses or otherwise avoid using their medications because of cost. We sought to identify which elderly patients are at highest risk of restricting their medications because of cost, and how prescription coverage modifies this risk. DESIGN AND PARTICIPANTS: Cross sectional study from the 1995-1996 wave of the Survey of Asset and Health Dynamics Among the Oldest Old, a population-based survey of Americans age 70 years and older. MEASUREMENTS: Subjects were asked the extent of their prescription coverage, and whether they had taken less medicine than prescribed for them because of cost over the prior 2 years. We used bivariate and multivariate analyses to identify risk factors for medication restriction in subjects who lacked prescription coverage. Among these high-risk groups, we then examined the effect of prescription coverage on rates of medication restriction. MAIN RESULTS: Of 4,896 seniors who regularly used prescription medications, medication restriction because of cost was reported by 8% of subjects with no prescription coverage, 3% with partial coverage, and 2% with full coverage (P <.01 for trend). Among subjects with no prescription coverage, the strongest independent predictors of medication restriction were minority ethnicity (odds ratio [OR], 2.9 compared with white ethnicity; 95% confidence interval [95% CI], 2.0 to 4.2), annual income <$10,000 (OR, 3.8 compared with income > or =$20,000; 95% CI, 2.4 to 6.1), and out-of-pocket prescription drug costs >$100 per month (OR, 3.3 compared to costs < or =$20; 95% CI, 1.5 to 7.2). The prevalence of medication restriction in members of these 3 risk groups was 21%, 16%, and 13%, respectively. Almost half (43%) of subjects with all 3 risk factors and no prescription coverage reported restricting their use of medications. After multivariable adjustment, high-risk subjects with no coverage had 3 to 15 times higher odds of medication restriction than subjects with partial or full coverage (P <.01). CONCLUSIONS: Medication restriction is common in seniors who lack prescription coverage, particularly among certain vulnerable groups. Seniors in these high-risk groups who have prescription coverage are much less likely to restrict their use of medications. PMID- 11903759 TI - A physician-based voluntary reporting system for adverse events and medical errors. AB - OBJECTIVE: To create a voluntary reporting method for identifying adverse events (AEs) and potential adverse events (PAEs) among medical inpatients. DESIGN: Medical house officers asked their peers about obstacles to care, injuries or extended hospitalizations, and problems with medications that affected their patients. Two independent reviewers coded event narratives for adverse outcomes, responsible parties, preventability, and process problems. We corroborated house officers' reports with hospital incident reports and conducted a retrospective chart review. SETTING: The cardiac step-down, oncology, and medical intensive care units of an urban teaching hospital. INTERVENTION: Structured confidential interviews by postgraduate year-2 and -3 medical residents of interns during work rounds. MEASUREMENTS AND MAIN RESULTS: Respondents reported 88 events over 3 months. AEs occurred among 5 patients (0.5% of admissions) and PAEs among 48 patients (4.9% of admissions). Delayed diagnoses and treatments figured prominently among PAEs (54%). Clinicians were responsible for the greatest number of incidents (55%), followed by workers in the laboratory (11%), radiology (15%), and pharmacy (3%). Respondents identified a variety of problematic processes of care, including problems with diagnosis (16%), therapy (26%), and failure to provide clinical and support services (29%). We corroborated 84% of reported events in the medical record. Participants found voluntary peer reporting of medical errors unobtrusive and agreed that it could be implemented on a regular basis. CONCLUSIONS: A physician-based voluntary reporting system for medical errors is feasible and acceptable to front-line clinicians. PMID- 11903758 TI - Patient-centered processes of care and long-term outcomes of myocardial infarction. AB - OBJECTIVE: To examine whether patients' experiences with nontechnical aspects of care such as patient education and discharge planning are associated with long term outcomes. DESIGN: Observational cohort study. SETTING: Twenty-three New Hampshire hospitals during 1996 and 1997. PARTICIPANTS: Acute myocardial infarction (AMI) patients (N=2,272) enrolled prior to discharge. MEASUREMENTS: Surveys asking about problems with care and health were mailed to patients 1, 3, and 12 months after discharge. Patients were stratified into "worse" or "better" care groups on the basis of their hospital care problem score. Outcomes included self-reported overall health, physical health, mental health, chest pain, and shortness of breath. Other clinical measures were obtained from hospital discharge abstracts. MAIN RESULTS: The 1-, 3-, and 12-month surveys were returned by 1,346 (59.2%), 1,046 (46%), and 964 (42.4%) enrolled patients, respectively. The primary analytic cohort consisted of the 762 patients who completed both the 1- and 12-month surveys. After adjustment for postdischarge health status and other clinical factors, patients experiencing worse hospital care had lower ratings of overall health (48.4 vs 52.5 on 100-point scale; P=.02) and physical health (59.7 vs 68.4; P <.001), and were more likely to have chest pain (odds ratio [OR], 1.6; confidence interval [CI], 1.0 to 2.4; P=.04) 12 months after their AMI than other patients. However, differences in reports of chest pain were reduced if patients reporting worse hospital care had better experiences with subsequent ambulatory care. CONCLUSIONS: Patients' experiences with nontechnical processes of AMI hospital care are associated with long-term outcomes; however the association between a negative hospital experience and subsequent chest pain may be offset by more positive outpatient experiences. PMID- 11903760 TI - Specialists' and primary care physicians' participation in medicaid managed care. AB - OBJECTIVE: To compare specialist and primary care physician participation in California's Medicaid fee-for-service and managed care programs. DESIGN: Cross sectional survey. PARTICIPANTS: A probability sample stratified by county and by race of 962 specialist physicians and 713 primary care physicians practicing in the 13 largest counties in California in 1998. MEASUREMENTS AND ANALYSIS: We used physician self-report from mailed questionnaires to compare acceptance of new Medicaid and new Medicaid managed care patients by specialists versus primary care physicians and by physician demographics, practice setting, attitudes toward Medicaid patients, and attitudes toward Medicaid managed care. We analyzed results using logistic regression with data weighted to represent the total population of primary care and specialist physicians in the 13 counties. MAIN RESULTS: Specialists were as likely as primary care physicians to have any Medicaid patients in their practices (56% vs 56%; P=.9). Among physicians accepting any new patients, specialists were more likely than primary care physicians to be taking new Medicaid patients but were significantly more likely to limit their acceptance to only Medicaid fee-for-service patients. Thus, specialists were much less likely than primary care physicians to accept new Medicaid managed care patients. After controlling for physician demographics, practice settings, and attitudes toward Medicaid patients and Medicaid managed care, specialists remained much less likely to accept new Medicaid managed care patients. CONCLUSIONS: Expansion of Medicaid managed care may decrease access to specialists as specialists were less likely to accept new Medicaid managed care patients compared to Medicaid fee-for-service patients. Any decrease in access may be mitigated if states are able to contract with group model HMOs and to recruit minority physicians. PMID- 11903761 TI - Attitudes toward colorectal cancer screening tests. AB - OBJECTIVE: To examine patient and physician preferences in regard to 5 colorectal cancer screening alternatives endorsed by a 1997 expert panel, determine the impact of patient and physician values regarding certain test features on screening preference, and assess physicians' perceptions of patients' values. DESIGN: Cross-sectional survey. SETTING: A general internal medicine practice at an academic medical center in 1998. PARTICIPANTS: Patients (N=217; 76% response rate) and physicians (N=39; 87% response rate) at the study setting. MEASUREMENTS AND MAIN RESULTS: Patients preferred fecal occult blood testing (43%) or colonoscopy (40%). In patients for whom accuracy was the most important test feature, colonoscopy (62%) was the preferred screening method. Patients for whom invasive test features were more important preferred fecal occult blood testing (76%; P <.001). Patients and physicians were similar in their values regarding the various test features. However, there was a significant difference between physicians' perceptions of which test features were important to patients compared with the patients' actual responses (P <.001). The largest discrepancy was for accuracy (patient actual 54% vs physician opinion 15%) and discomfort (patient actual 15% vs physician opinion 64%). CONCLUSIONS: Patients have distinct preferences for colorectal cancer screening tests that are associated with the importance placed on certain test features. Physicians incorrectly perceive those factors that are important to patients. Physicians should incorporate patient values in regard to certain test features when discussing colorectal cancer screening with their patients and when eliciting their screening preferences. PMID- 11903762 TI - Prostate cancer screening practices and beliefs. AB - OBJECTIVE: To examine primary care physician prostate-specific antigen (PSA) testing and prostate cancer screening beliefs, practices, and trends over time. DESIGN: Longitudinal physician survey. SETTING: Community and academic primary care practices in a major East Coast city. PARTICIPANTS: Physicians with > or =100 patients enrolled in an HMO serving 90,000 patients were surveyed in 1993 and 1998 regarding prostate cancer screening beliefs and practices. In 1993, 176 physicians (76%) completed the survey. In 1998, 76% of the 1993 respondents responded to a second survey. Associations between and changes over time in beliefs and self-reported PSA testing were analyzed. MEASUREMENTS AND MAIN RESULTS: Physicians reported ordering PSA tests in 73% of health maintenance exams (HMEs) in 1993 and 81% of HMEs in 1998. PSA testing significantly increased between 1993 and 1998 with 43% of physicians reporting increased testing and only 13% reporting reduced testing. Between 1993 and 1998, physician attitudes favoring PSA testing increased although less than half of physicians believed that aggressive early treatment improved patient outcomes. Community versus academic practice location was also associated with PSA testing. CONCLUSIONS: Physicians reported high and increasing rates of PSA testing from 1993 to 1998 and more favorable attitudes toward PSA testing. Despite conflicting expert recommendations and a lack of consistent, high-quality supporting evidence, PSA screening appears to be increasingly considered a standard of care by practicing physicians. However, beliefs other than improved patient outcomes due to screening may be the primary drivers of increased PSA testing. PMID- 11903763 TI - Resident utilization of information technology. AB - OBJECTIVE: To determine if a simple educational intervention can increase resident physician literature search activity. DESIGN: Randomized controlled trial. SETTING: University hospital-based internal medicine training program. PATIENTS/PARTICIPANTS: Forty-eight medical residents rotating on the general internal medicine service. INTERVENTIONS: One-hour didactic session, the use of well-built clinical question cards, and practical sessions in clinical question building. MEASUREMENTS AND MAIN RESULTS: Objective data from the library information system that included the number of log-ons to medline, searching volume, abstracts viewed, full-text articles viewed, and time spent searching. Median search activity as measured per person per week (control vs intervention): number of log-ons to medline (2.1 vs 4.4, P <.001); total number of search sets (24.0 vs 74.2, P <.001); abstracts viewed (5.8 vs 17.7, P=.001); articles viewed (1.0 vs 2.6, P=.005); and hours spent searching (0.8 vs 2.4, P <.001). CONCLUSIONS: A simple educational intervention can markedly increase resident searching activity. PMID- 11903764 TI - Ultrasonography performed by primary care residents for abdominal aortic aneurysm screening. AB - A prospective pilot study was undertaken to assess a protocol to educate primary care residents in how to personally perform ultrasonography for abdominal aortic aneurysm screening. Resident exams were proctored by a primary care physician trained in ultrasonography and were scored on the level of competence in doing the examination. Patients had ultrasound performed by a resident, followed by repeat examination by the vascular lab. Primary care resident abdominal aortic imaging was achieved in 79 of 80 attempts. Four abdominal aortic aneurysms were identified. There were 75 normal examinations; resident ultrasonography results were consistent with the results of the vascular lab. Ten residents achieved an abdominal aortic ultrasound-independent competence level after an average of 3.4 proctored exams. The main outcome of this study is that a primary care resident, with minimal training in ultrasonography imaging, is able to rapidly learn the technique of ultrasonography imaging of the abdominal aorta. PMID- 11903766 TI - Impact of a first-year primary care experience on residency choice. AB - We designed a retrospective cohort study of first-year medical students to assess the impact of a community-based primary care course, Introduction to Primary Care (IPC), on residency choice. In the group that took IPC (n=282), 48.2% entered generalist residencies (internal medicine, pediatrics, family medicine, or medicine/pediatrics), compared to 38.2% in the group that wanted IPC (n=398) and 39.6% in the group that did not want IPC (n=245). Controlling for gender, students who took IPC had a 40% higher odds of selecting a generalist residency than those who wanted to take IPC (odds ratio [OR], 1.42; 95% confidence interval [CI], 1.04 to 1.95). There was no difference between those who wanted IPC and those who did not (OR, 1.08; CI, 0.78 to 1.52). The community-based primary care experience was positively associated with students' selection of generalist residencies. PMID- 11903765 TI - Initiation and continuation of newer antiretroviral treatments among medicaid recipients with AIDS. AB - OBJECTIVE: To examine initiation of newer antiretroviral treatments across sociodemographic subgroups during the 3 years following the introduction of these treatments, and explore persistence on treatment and its association with patient characteristics. DESIGN: Merged Medicaid paid claims and HIV/AIDS surveillance data were used to analyze use of protease inhibitors (PIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) over time. Survival analysis techniques were used to analyze initiation of PI/NNRTI use. Ordinary least squares and logistic regression were used to determine predictors of persistence on PI/NNRTI therapy. SETTING AND PARTICIPANTS: The study population consisted of 2,459 New Jersey non-HMO adult Medicaid beneficiaries with AIDS, identified through a match between HIV/AIDS Registry and Medicaid files. Their PI/NNRTI use was followed from March 1996, when the first PI was licensed, to the end of 1998. MEASUREMENTS AND MAIN RESULTS: African Americans initiated treatment on average 8 months later than non-Hispanic whites; initiation of treatment was also slower for injection drug users and for those who did not receive case management through a Medicaid waiver program. These bivariate findings were confirmed with a multivariate time to-treatment analysis using proportional hazards regression. Among those initiating PI/NNRTI use, 35% had discontinued it by the end of follow-up. Bivariate analyses of treated individuals found that PI/NNRTI use as a proportion of follow-up time was lower for African Americans and Hispanics, and higher for older individuals and for those receiving case management through a Medicaid waiver program, while injection drug use history was not associated with persistence. These findings were confirmed by a regression analysis, which found that controlling for other characteristics, African-American race, and Hispanic ethnicity were each associated with a significant 8% reduction in the proportion of time on PI/NNRTIs following initiation of treatment. Alternative approaches for modeling persistence produced similar results. CONCLUSIONS: Results suggest that consistent longitudinal use is difficult for many patients. Persistence of use was lower for minority beneficiaries despite comparable coverage for pharmacy and other health services through Medicaid. Our findings suggest the need to examine nonfinancial barriers to appropriate use of highly active antiretroviral therapy, and to develop and test programmatic strategies for supporting patients in remaining on these regimens consistently. PMID- 11903767 TI - Medicare prescription coverage and congressional gridlock. PMID- 11903768 TI - Cancer screening in primary care. Are we communicating? PMID- 11903769 TI - Processes and outcomes in acute myocardial infarction. PMID- 11903771 TI - Specialty training and specialization among physicians who treat HIV/AIDS in the United States. AB - OBJECTIVE: To assess the association of specialty training and experience in the care of HIV disease with HIV-specific knowledge, referral patterns, and HIV related education activities. DESIGN: Cross-sectional survey. SETTING: The United States. PARTICIPANTS: Physicians caring for patients in the HIV Costs and Service Utilization Study, a study of a probability sample of HIV-infected individuals in the United States. MEASUREMENTS AND MAIN RESULTS: Measures included physicians' reports of specialty training and HIV caseload, scores on an HIV-specific knowledge test, referral patterns, and attendance rates at HIV-related educational activities. Approximately 72% (379) of the eligible physicians completed a survey. Of these, 152 (40%) had infectious disease (ID) training, and 213 (56%) were generalists; 4% of ID-trained physicians and 37% of generalist physicians did not consider themselves HIV experts. The median current caseloads were 150 and 200 patients for ID experts and generalist experts, respectively. In contrast, the median caseload for non-expert generalists was 5. Mean scores on the knowledge scale were similar for ID and generalist experts (9.0 items correct out of 11 vs 8.5; P=not significant), but lower for generalist non-experts (6.5 items correct; P <.01). Experts had attended more local and national HIV meetings than non-experts (9.3 vs 2.7; P <.01, and 2.3 vs.40; P <.01, respectively) in the past year. Fewer ID experts ever referred than generalist experts (13.0% vs 27.3%; P=.01). In multivariable models that included specialty training and caseload, physicians with caseloads of 20 to 49 and >50 were more likely to have a high knowledge score (defined as 80% or more correct, odds ratio [OR], 2.8; P=.04 and OR, 5.7; P <.001, respectively), and the effect of specialty was attenuated (OR, 2.7; P=.02 decreased from OR, 7.8; P <.001 in a model without caseload). In the models predicting referral practices, both experience (OR,.25; P <.01 and OR,.17; P <.01 for caseloads of 20 to 49 and >50, respectively) and specialty (OR,.19; P <.01 and OR,.09; P <.01 for generalist and ID experts, respectively) were significant. CONCLUSIONS: In a national sample of physicians, HIV-specific knowledge was more strongly associated with HIV caseload than with specialty training. In addition, although referral practices were related to both experience and specialty, generalist experts and ID physicians reported similar behaviors. This suggests that generalist physicians, through clinical experience and self-education, can develop specialized knowledge in HIV care. PMID- 11903770 TI - How well do clinicians estimate patients' adherence to combination antiretroviral therapy? AB - OBJECTIVE: Adherence to combination antiretroviral therapy is critical for clinical and virologic success in HIV-infected patients. To combat poor adherence, clinicians must identify nonadherent patients so they can implement interventions. However, little is known about the accuracy of these assessments. We sought to describe the accuracy of clinicians' estimates of patients' adherence to combination antiretroviral therapy. SETTING: Public HIV clinic. DESIGN: Prospective cohort study. During visits, we asked clinicians (nurse practitioners, residents and fellows, and their supervising attending physicians) to estimate the percentage of antiretroviral medication taken by patients over the last 4 weeks and predicted adherence over the next 4 weeks. Adherence was measured using electronic monitoring devices, pill counts, and self-reports, which were combined into a composite adherence measure. PATIENTS AND PARTICIPANTS: Clinicians estimated 464 episodes of adherence in 82 patients. RESULTS: Among the 464 adherence estimates, 264 (57%) were made by principal care providers (31% by nurse practitioners, 15% by fellows, 6% by residents, and 5% by staff physicians) and 200 (43%) by supervising attending physicians. Clinicians' overestimated measured adherence by 8.9% on average (86.2% vs 77.3%). Greater clinician inaccuracy in adherence prediction was independently associated with higher CD4 count nadir (1.8% greater inaccuracy for every 100 CD4 cells, P=.005), younger patient age (3.7% greater inaccuracy for each decade of age, P=.02), and visit number (P=.02). Sensitivity of detecting nonadherent patients was poor (24% to 62%, depending on nonadherence cutoff). The positive predictive value of identifying a patient as nonadherent was 76% to 83%. CONCLUSIONS: Clinicians tend to overestimate medication adherence, inadequately detect poor adherence, and may therefore miss important opportunities to intervene to improve antiretroviral adherence. PMID- 11903773 TI - How are patients' specific ambulatory care experiences related to trust, satisfaction, and considering changing physicians? AB - CONTEXT: Few data are available regarding the consequences of patients' problems with interpersonal aspects of medical care. OBJECTIVE: To assess the relationships between outpatient problem experiences and patients' trust in their physicians, ratings of their physicians, and consideration of changing physicians. We classified as problem experiences patients' reports that their physician does not always 1) give them enough time to explain the reason for the visit, 2) give answers to questions that are understandable, 3) take enough time to answer questions, 4) ask about how their family or living situation affects their health, 5) give as much medical information as they want, or 6) involve them in decisions as much as they want. DESIGN: Telephone survey during 1997. PARTICIPANTS: Patients (N=2,052; 58% response) insured by a large national health insurer. MEASUREMENTS: Patient trust, overall ratings of physicians, and having considered changing physicians. RESULTS: Most patients (78%) reported at least 1 problem experience. In multivariable analyses, each problem experience was independently associated with lower trust (all P <.001) and 5 of 6 with lower overall ratings (P <.001). Three problem experiences were independently related to considering changing physicians: physicians not always giving answers to questions that are understandable (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.3 to 3.0), not always taking enough time to answer questions (OR, 3.3; 95% CI, 2.2 to 5.2), and not always giving enough medical information (OR, 4.0; 95% CI, 2.4 to 6.6). CONCLUSIONS: Problem experiences in the ambulatory setting are strongly related to lower trust. Several are also associated with lower overall ratings and with considering changing physicians, particularly problems related to communication of health information. Efforts to improve patients' experiences may promote more trusting relationships and greater continuity and therefore should be a priority for physicians, educators, and health care organizations. PMID- 11903772 TI - Screening for diabetes in an outpatient clinic population. AB - BACKGROUND: Opportunistic disease screening is the routine, asymptomatic disease screening of patients at the time of a physician encounter for other reasons. While the prevalence of unrecognized diabetes in community populations is well known, the prevalence in clinical populations is unknown. OBJECTIVE: To describe the prevalence, predictors, and clinical severity of unrecognized diabetes among outpatients at a major medical center. DESIGN AND SETTING: A cross-sectional observational study at the Durham Veterans Affairs Medical Center. SUBJECTS: Outpatients without recognized diabetes (N=1,253). METHODS: We screened patients for diabetes by using an initial random Hemoglobin A1c (HbA1c) measurement, and then obtaining follow-up fasting plasma glucose (FPG) for all subjects with HbA1c > or =6.0%. A case of unrecognized diabetes was defined as either HbA1c > or =7.0% or FPG > or =7 mmol/L (126 mg/dL). Height and weight were obtained for all subjects. We also obtained resting blood pressure, fasting lipids, and urine protein in subjects with HbA1c > or =6.0%. RESULTS: The prevalence of unrecognized diabetes was 4.5% (95% confidence interval [CI], 3.4 to 5.7). Factors associated with unrecognized diabetes were the diagnosis of hypertension (adjusted odds ratio [OR], 2.5; P=.004), weight >120% of ideal (adjusted OR, 2.2; P=.02), and history of a parent or sibling with diabetes (adjusted OR, 1.7; P=.06). Having a primary care provider did not raise or lower the risk for unrecognized diabetes (P=.73). Based on the new diagnosis, most patients (61%) found to have diabetes required a change in treatment either of their blood sugar or comorbid hypertension or hyperlipidemia in order to achieve targets recommended in published treatment guidelines. Patients reporting a primary care provider were no less likely to require a change in treatment (P=.20). CONCLUSIONS: If diabetes screening is an effective intervention, opportunistic screening for diabetes may be the preferred method for screening, because there is substantial potential for case-finding in a medical center outpatient setting. A majority of patients with diabetes diagnosed at opportunistic screening will require a change in treatment of blood sugar, blood pressure, or lipids to receive optimal care. PMID- 11903776 TI - Early introduction of an evidence-based medicine course to preclinical medical students. AB - Evidence-based Medicine (EBM) has been increasingly integrated into medical education curricula. Using an observational research design, we evaluated the feasibility of introducing a 1-month problem-based EBM course for 139 first-year medical students at a large university center. We assessed program performance through the use of a web-based curricular component and practice exam, final examination scores, student satisfaction surveys, and a faculty questionnaire. Students demonstrated active involvement in learning EBM and ability to use EBM principles. Facilitators felt that students performed well and compared favorably with residents whom they had supervised in the past year. Both faculty and students were satisfied with the EBM course. To our knowledge, this is the first report to demonstrate that early introduction of EBM principles as a short course to preclinical medical students is feasible and practical. PMID- 11903774 TI - Patient protection and risk selection: do primary care physicians encourage their patients to join or avoid capitated health plans according to the patient's health status? AB - BACKGROUND: Individual physicians who are paid prospectively, as in capitated health plans, might tend to encourage patients to avoid or to join these plans according to the patient's health status. Though insurance risk selection has been well documented among organizations paid on a prospective basis, such physician-level risk selection has not been studied. OBJECTIVE: To assess physician reports of risk selection in capitated health plans and explore potentially related factors. DESIGN AND PARTICIPANTS: National mailed survey of primary care physicians in 1997-1998, oversampling physicians in areas with more capitated health plans. RESULTS: The response rate was 63% (787 of 1,252 eligible recipients). Overall, 44% of physicians reported encouraging patients either to join or to avoid capitated health plans according to the patients' health status: 40% encouraged more complex and ill patients to avoid capitated plans and 23% encouraged healthier patients to join capitated plans. In multivariable models, physicians with negative perceptions of capitated plan quality, with more negative experiences in capitated plans, and those who knew at each patient encounter how they were being compensated had higher odds of encouraging sicker patients to avoid capitated plans (odds ratios, 2.0, 2.2, and 2.0; all confidence intervals >1). CONCLUSIONS: Many primary care physicians report encouraging patients to join or avoid capitated plans according to the patient's health status. Although these physicians' recommendations might be associated primarily with concerns about quality, they can have the effect of insulating certain health plans from covering sicker and more expensive patients. PMID- 11903775 TI - Cross-cultural similarities and differences in attitudes about advance care planning. AB - OBJECTIVE: Culture may have an important impact on a patient's decision whether to perform advance care planning. But the cultural attitudes influencing such decisions are poorly defined. This hypothesis-generating study begins to characterize those attitudes in 3 American ethnic cultures. DESIGN: Structured, open-ended interviews with blinded content analysis. SETTING: Two general medicine wards in San Antonio, Texas. PATIENTS: Purposive sampling of 26 Mexican American, 18 Euro-American, and 14 African-American inpatients. MEASUREMENTS AND MAIN RESULTS: The 3 groups shared some views, potentially reflecting elements of an American core culture. For example, majorities of all groups believed "the patient deserves a say in treatment," and "advance directives (ADs) improve the chances a patient's wishes will be followed." But the groups differed on other themes, likely reflecting specific ethnic cultures. For example, most Mexican Americans believed "the health system controls treatment," trusted the system "to serve patients well," believed ADs "help staff know or implement a patient's wishes," and wanted "to die when treatment is futile." Few Euro Americans believed "the system controls treatment," but most trusted the system "to serve patients well," had particular wishes about life support, other care, and acceptable outcomes, and believed ADs "help staff know or implement a patient's wishes." Most African Americans believed "the health system controls treatment," few trusted the system "to serve patients well," and most believed they should "wait until very sick to express treatment wishes." CONCLUSION: While grounded in values that may compose part of American core culture, advance care planning may need tailoring to a patient's specific ethnic views. PMID- 11903778 TI - Talking about money: how primary care physicians respond to a patient's question about financial incentives. AB - Patients sometimes express concern about the influence of "perverse" financial incentives on their care. We recruited a convenience sample of 101 primary care physicians and obtained information on their compensation. Then we audiotaped them as they role-played a response to a videotaped mock patient who asked them how they were paid and how their method of compensation affected clinical decisions. Overall, 36% of the physicians did not give enough information in their role-play response to allow an independent determination of how they were paid. Adopting a broad spectrum of attitudes and approaches, nearly every physician avoided discussing the role of incentives and stressed instead that he or she could be trusted under any circumstance. PMID- 11903777 TI - Beyond the examination room: primary care performance and the patient-physician relationship for low-income women. AB - OBJECTIVE: To assess whether primary care performance of low-income women's primary care delivery sites is associated with the strength of their relationships with their physicians. DESIGN: Random-digit-dial and targeted household telephone survey of a population-based sample. SETTING: Washington, D.C. census tracts with > or =30% of households below 200% of federal poverty threshold. PARTICIPANTS: Women over age 40 (N=1,205), 82% of whom were African American. MEASUREMENTS AND MAIN RESULTS: The response rate was 85%. Primary care performance was assessed using women's ratings of their systems' accessibility (organizational, geographic, and financial), continuity, comprehensiveness, and coordination. Respondents' ratings of trust in their physicians, communication with their physicians, and compassion shown by their physicians were used to operationalize the patient-physician relationship. Controlling for population and insurance characteristics, 4 primary care features were positively associated with women's trust in and communication with their physicians: continuity with a single clinician, organizational accessibility of the practice, comprehensive care, and coordination of specialty care services. Better organizational access, but not geographic or financial access, was associated with greater levels of trust, compassion, and communication (odds ratios [ORs], 3.2, 7.4, and 6.9, respectively; P < or =.01). Women who rated highest their doctor's ability to take care of all of their health care needs (highest level of comprehensiveness) had 11 times the odds of trusting their physician (P < or =.01) and 6 times the odds of finding their physicians compassionate and communicative (P < or =.01), compared to those with the lowest level of comprehensiveness. CONCLUSIONS: Primary care delivery sites organized to be more accessible, to link patients with the same clinician for their visits, to provide for all of a woman's health care needs, and to coordinate specialty care services are associated with stronger relationships between low-income women and their physicians. Primary care systems that fail to emphasize these features of primary care may jeopardize the clinician-patient relationship and indirectly the quality of care and health outcomes. PMID- 11903779 TI - Trust, distrust and trustworthiness. PMID- 11903780 TI - Who cares for AIDS? PMID- 11903781 TI - A call for papers: Community-based participatory research. PMID- 11903782 TI - Guidelines for anticoagulation therapy for venous thromboembolism. PMID- 11903783 TI - Editorial: Continuing professional development. PMID- 11903784 TI - Treating obesity: a follow-up study. Can the stages of change model be used as a postal screening tool? AB - AIMS: We have previously shown that although a postal questionnaire based on the stages of change model (SCQ) failed to distinguish outpatients most likely to lose weight, it appeared to influence attendance rates. We therefore audited attendance upon receiving a pre-appointment SCQ in the post and compared this to previous standard practice in 1996. METHODOLOGY AND RESULTS: Seventy-eight obese outpatients (BMI 36.7 +/- 6.7 kg m-2, age 43 +/- 15 years) (mean +/- SD) were included. Twenty-nine per cent of patients failed to return an SCQ and were not sent an appointment and therefore did not block dietetic time. Eleven per cent returned an SCQ but did not attend (DNA) visit 1, whilst 21% attended visit 1 but DNA visit 2. Thirty-nine per cent attended both appointments and lost a significant amount of weight (105 +/- 23 vs. 103.2 +/- 23 kg, P < 0.0001) in 4 weeks. There was no difference in SCQ results between groups. Overall attendance rate at initial and follow-up appointments increased by 11%, while DNA rates fell by 20% compared with the 1996 audit. CONCLUSION: The SCQ has sifted out one in four patients who previously DNA an initial dietetic outpatient appointment. This has reduced waiting-list time as appointments are only book on return on the SCQ form, increased effective use of dietetic time through increased attendance rates, and lowered the DNA rate of follow-up appointments. PMID- 11903785 TI - Probability of assertive behaviour, interpersonal anxiety and self-efficacy of South African registered dietitians. AB - OBJECTIVES: There is little information on the probability of assertive behaviour, interpersonal anxiety and self-efficacy in the literature regarding dietitians. The objective of this study was to establish baseline information of these attributes and the factors affecting them. METHOD: Questionnaires collecting biographical information and self-assessment psychometric scales measuring levels of probability of assertiveness, interpersonal anxiety and self efficacy were mailed to 350 subjects, who comprised a random sample of dietitians registered with the Health Professions Council of South Africa. RESULTS: Forty one per cent (n=145) of the sample responded. Self-assessment inventory results were compared to test levels of probability of assertive behaviour, interpersonal anxiety and self-efficacy. The inventory results were compared with the biographical findings to establish statistical relationships between the variables. The hypotheses were formulated before data collection. CONCLUSION: Dietitians had acceptable levels of probability of assertive behaviour and interpersonal anxiety. The probability of assertive behaviour was significantly lower than the level noted in the literature and was negatively related to interpersonal anxiety and positively related to self-efficacy. PMID- 11903786 TI - Dietary restraint in relation to nutrient intake, physical activity and iron status in adolescent females. AB - AIM: To investigate the prevalence of dietary restraint in a female adolescent population, and to examine the nutritional consequences of dietary restraint and its implications for iron status. METHODS: A total of 64 adolescent females, aged 14-15 years, were recruited from two all-girl schools in central London. Nutrient intake, body weight, physical activity and iron status were measured. Findings were compared between three groups of subjects classified by dietary restraint. RESULTS: Adolescents with a higher BMI percentile were more likely to be highly restrained. Scores on the dietary restraint psychometric measures were comparable with other UK studies in this age group. Energy intake was inversely related to dietary restraint (mean energy intake (SE) for each restraint group were: low 8.99 MJ (0.48), medium 7.98 MJ (0.22) and high 7.35 MJ (0.39) P < 0.05); however, a corresponding relationship between dietary restraint and reduced micronutrient intakes was not found. Highly restrained eaters obtained more of their energy intake from bread, fruit and cheese and less from meat, meat products and confectionery. Levels of physical activity were not significantly different between the dietary restraint groups. There was a poor relationship between reported energy intake and estimated energy expenditure. Haematological parameters of iron status were similar across the restraint groups. CONCLUSIONS: Dietary restraint was exercised by the consumption of a "healthy eating diet". Dieting was not related to a lower iron status; however, the low dietary iron intake and poor iron status of the whole sample is of concern. PMID- 11903787 TI - Breast-feeding knowledge and attitudes of teenage mothers in Liverpool. AB - OBJECTIVES: To determine the knowledge and attitudes of teenage mothers towards breast-feeding. DESIGN: A questionnaire of teenage (< 20 years) and non-teenage (> or = 20 years) primigravidae attending the antenatal care services at the Liverpool Women's Hospital, during the period April-May 2000. RESULTS: Forty teenage primigravidae and 40 non-teenage primigravidae registered for the survey. Teenagers had poorer knowledge about breast-feeding than the non-teenagers, and fewer teenagers considered breast milk the best food for their baby. More teenagers than non-teenagers planned to bottle feed [23 (57.5%) vs. 9 (22.5%), P=0.002]. Only one teenager had knowledge about colostrum. Teenagers were more often single, had a lower level of education, higher unemployment, higher smoking frequency and less contact with a person who had previously breast-fed. CONCLUSION: Teenage primigravidae have poor knowledge regarding breast-feeding compared with non-teenage primigravidae. A greater proportion of teenagers opted not to breast-feed compared with non-teenagers. Health education classes stressing the importance of breast-feeding should be emphasized in antenatal teenage clinics. More research is needed to understand how to improve the knowledge and motivation of adolescent girls to breast feed. PMID- 11903788 TI - Dietary calcium in galactosaemia. AB - The diets of 19 galactosaemic patients were assessed for calcium and phosphorous intake. Despite the use of infant soya formula or calcium-supplemented soya milk the reference nutrient intake (RNI) for calcium was only met in 26% of the group; all patients met > 100% of the RNI for phosphorous. The regular assessment of the diets of galactosaemia patients is recommended as the milk-free nature of the diet can lead to an inadequate calcium intake. Lactose-free calcium supplements should be prescribed if the diet alone is inadequate. PMID- 11903789 TI - Impact of stepped verbal and written reinforcement of fluid balance advice within an outpatient haemodialysis unit: a pilot study. AB - BACKGROUND: The present pilot study assesses a programme of verbal and written fluid balance advice within an out-patient haemodialysis unit. METHODS: Twenty one haemodialysis patients were followed over three separate 6-week periods. Each 6-week period assessed dietetic and nursing educational input using a stepped approach with interdialytic weight gain (IDWG) as a barometer of patient compliance. Weight gain was statistically examined using the non-parametric Friedman test. RESULTS: Forty-eight per cent of the sample group demonstrated an overall improvement in mean weight gain; however, this was not statistically significant (P=0.504). CONCLUSION: The pilot study suggests that further studies with greater numbers and a control group would have a useful contribution to make in this field. PMID- 11903790 TI - An audit of nutritional care delivered to elderly inpatients in community hospitals. AB - BACKGROUND: There is continuing concern over the lack of attention to the nutritional needs of older people in hospitals. A 2-year audit project was undertaken to examine the nutritional care of inpatients in Leicestershire Community Hospitals. METHOD: The methods used included analysis of menu cycles; observation of meal and drink provision, wastage, supplement usage and portion sizes; and patient satisfaction questionnaire examination. RESULTS: Patient menus were nutritionally inadequate for energy, fibre and vitamin D, and protein levels were variable. The percentage of meal wastage and inadequate portion sizes were of concern. Patient satisfaction results were overall positive. CONCLUSION: Patient energy intakes are a major concern with low calorie provision from menus exacerbated by a deficit in recommended portion sizes and a high percentage of meal wastage. Routine audits need to be implemented to monitor both portion size and meal wastage, and to address patient satisfaction issues to improve the overall intakes of patients. Multidisciplinary team input is required to address the above issues and additional recommendations to promote nutrition as a key component in clinical care. PMID- 11903791 TI - The methodology of nutritional screening and assessment tools. AB - BACKGROUND: The aim of this paper was to give a critical appraisal of the methodology of nutritional screening and assessment tools. METHODS: A literature search identified 44 tools. Each tool was assessed in relation to details of its application, method of derivation and evaluation of its performance. RESULTS: The findings indicate that tools were published with insufficient details regarding their intended use and method of derivation, and with an inadequate assessment of their effectiveness. An appraisal of these features judged that no one tool satisfied a set of criteria regarding scientific merit. CONCLUSION: There is thus a need to ensure that nutritional screening and assessment tools are developed using procedures based on good design and sound statistical practice. This paper suggests that a unified approach using multivariate techniques could make a significant contribution to this process. PMID- 11903793 TI - International perspectives. PMID- 11903795 TI - Radiation damage of water in environmental scanning electron microscopy. AB - Specimen damage from the electron beam poses a considerable problem with electron microscopy. This damage is particularly acute in environmental scanning electron microscopy (ESEM) for two reasons. Firstly, owing to its ability to stabilise insulating and hydrated specimens, ESEM lends itself to polymeric and biological materials that are typically highly beam-sensitive. Secondly, water acts as a source of small, highly mobile free radicals, which accelerate specimen degradation. By taking the results of single-particle simulations of electron water interactions, we determine the concentration of reactive species in a water specimen under ESEM conditions. We consider 12 species, which are produced in a Gaussian distribution, and annihilate according to a second-order reaction scheme. Self-diffusion along the concentration gradient is also modelled. We find that the dominant reactive species is the hydroxyl (.OH) radical. Annihilation of this species is suppressed due to the lower concentration of reactants. The relatively stable hydrogen peroxide is also found at large concentrations. By comparing two beam energies, 5 and 25 keV, we find a drastic increase in the quantities of reactive species produced with beam energy. The longer range of 25 keV primary electrons spreads reactive species over a wider region, which then decay far more slowly. PMID- 11903796 TI - Detection of mitochondrial DNA in living animal cells with fluorescence microscopy. AB - The detection of mitochondrial DNA (mtDNA) in living human cells could be useful for understanding mitochondrial behaviour during cellular processes and pathological mtDNA depletions. However, until now, human mtDNA has not been visualized in living cells with fluorescence microscopy, although it has been easily detected in organisms with larger mtDNA. Previous reports have stated that mtDNA staining results in homogeneous fluorescence of mitochondria or that animal mitochondria are refractory to DAPI staining. This paper shows that mtDNA of cultured green monkey kidney CV-1 can be stained using a very low concentration of DAPI, then detected by a cooled Photometrics CCD camera with 14-bit resolution detection. Indeed, under these conditions CV-1 cells have small fluorescent spots in the cytoplasm that colocalize with mitochondria, even after mitochondrial movements, uncoupling by carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone and swelling. These observations have been reproduced for the human fibroblast foreskin cell line HS68. These results and known properties of DAPI as a specific DNA stain strongly suggest that mtDNA can be detected and visualized by fluorescence microscopy in human living cells, with potential developments in the study of mtDNA in normal and pathological situations. PMID- 11903797 TI - Microscopical investigations on the epicuticle of mammalian keratin fibres. AB - The existence of a thin chemically resistant layer, the epicuticle, close to the surfaces of all undamaged mammalian keratin fibres has been known since 1916. The identification of such a specific structure within the fibre cuticle has remained elusive. Now, through transmission electron microscope investigations of stained transverse sections of hairs from various animal species, the epicuticle has been tentatively identified as a sharply defined and continuous layer approximately 13 nm thick covering the entire outwardly facing intracellular surface of every cuticle cell. The staining behaviour of the epicuticle leads one to suppose that it is rich in cystine and that thioester-bound lipids might be present within its bulk. With the atomic force microscope it was established that the undamaged outer surface of all mammalian keratin fibres, even including those from the monotremes, were longitudinally striated. The lateral spacing of the striations was always in the range 0.29-0.39 microm. Striations only occurred on the freely exposed outer surfaces of the original undamaged fibres; evidently arising by some, as yet undefined, interaction in the follicle with the cuticle of the inner root sheath. By stripping off fatty acids known to be covalently attached to the fibre's outer surface, the striations were shown to reflect a corresponding irregularity of the epicuticle's surface. PMID- 11903798 TI - Atomic force microscopy of the erythrocyte membrane skeleton. AB - The atomic force microscope was used to examine the cytoplasmic surface of untreated as well as fixed human erythrocyte membranes that had been continuously maintained under aqueous solutions. To assess the effects of drying, some membranes were examined in air. Erythrocytes attached to mica or glass were sheared open with a stream of isotonic buffer, which allowed access to the cytoplasmic membrane face without exposing cells to non-physiological ionic strength solutions. Under these conditions of examination, the unfixed cytoplasmic membrane face revealed an irregular meshwork that appeared to be a mixture largely of triangular and rectilinear openings with mesh sizes that varied from 35 to 100 nm, although few were at the upper limit. Fixed ghosts were similar, but slightly more contracted. These features represent the membrane skeleton, as when the ghosts were treated to extract spectrin and actin, these meshworks were largely removed. Direct measurements of the thickness of the membrane skeleton and of the lateral dimensions of features in the images suggested that, especially when air dried, spectrin can cluster into large, quite regularly distributed aggregates. Aggregation of cytoskeletal components was also favoured when the cells were attached to a polylysine-treated substrate. In contrast, the membrane skeletons of cells attached to substrates rendered positively charged by chemical derivatization with a cationic silane were much more resistant to aggregation. As steps were taken to reduce the possibility of change of the skeleton after opening the cells, the aggregates and voids were eliminated, and the observed structures became shorter and thinner. Ghosts treated with Triton X-100 solutions to remove the bilayer revealed a meshwork having aggregated components resembling those seen in air. These findings support the proposition that the end-to-end distance of spectrin tetramers in the cell in the equilibrium state is much shorter than the contour length of the molecule and that substantial rearrangements of the spectrin-actin network occur when it is expanded by low ionic strength extraction from the cell. This study demonstrates the applicability of AFM for imaging the erythrocyte membrane skeleton at a resolution that appears adequate to identify major components of the membrane skeleton under near-physiological conditions. PMID- 11903799 TI - Bias of a length density estimator based on vertical projections. AB - Attention is paid to the stereological estimator of the length density of lineal features in three-dimensional space. In Gokhale (J. Microsc. (1990) 159, 133 141), the estimator based on measurements performed on a projection of the content of vertical slices with a given thickness Delta was derived. The aim of this paper is to show that using five vertical slices with systematically chosen orientations yields a reliable result, i.e. the bias of the estimator is smaller than 5%. In the case of a choice of the vertical axis such that most lineal features are not perpendicular or nearly perpendicular to the vertical axis, a reliable result can be obtained using only three systematic orientations of vertical slices. PMID- 11903800 TI - Tissue shrinkage and unbiased stereological estimation of particle number and size. AB - This paper is a review of the stereological problems related to the unbiased estimation of particle number and size when tissue deformation is present. The deformation may occur during the histological processing of the tissue. It is especially noted that the widely used optical disector may be biased by dimensional changes in the z-axis, i.e. the direction perpendicular to the section plane. This is often the case when frozen sections or vibratome sections are used for the stereological measurements. The present paper introduces new estimators to be used in optical fractionator and optical disector designs; the first is, as usual, the simplest and most robust. Finally, it is stated that when tissue deformation only occurs in the z-direction, unbiased estimation of particle size with several estimators is possible. PMID- 11903801 TI - Imaging the three orientation variants of the DO22 phase by 3D atom probe microscopy. AB - Three-phase NiAlV alloys were investigated using a three-dimensional atom probe. Ageing at 800 degrees C gives rise to the precipitation of two ordered phases within the supersaturated FCC solid solution, namely Ni3Al (L1(2) structure) and Ni3V (DO22 structure). The DO22 phase has three orientation variants which need to be identified in 3DAP images. It is shown that an appropriate choice of analysis site enables us to image the chemical order within both L1(2) and DO22 ordered phases and to distinguish the three orientation variants of the DO22 phase in reconstructed images. The lateral resolution of 3DAP in these experimental conditions was estimated through simple considerations to be less than 0.3 nm. PMID- 11903802 TI - Studies of the microstructure of polymer-modified bitumen emulsions using confocal laser scanning microscopy. AB - Polymer-modified bitumen emulsions present a safer and more environmentally friendly binder for enhancing the properties of roads. Cationic bitumen emulsion binders containing polymer latex were investigated using confocal laser scanning microscopy. The latex was incorporated into the bitumen emulsion by using four different addition methods and all emulsions were processed with a conventional colloid mill. The emulsion binder films were studied after evaporation of the emulsion aqueous phase. We show how the microstructure and distribution of the polymer varies within the bitumen binder depending on latex addition method, and that the microstructure of the binder remains intact when exposed to elevated temperature. It was found that a distinctly fine dispersion of polymer results when the polymer is blended into the bitumen before the emulsifying process (a monophase emulsion). In contrast, bi-phase emulsion binders produced by either post-adding the latex to the bitumen emulsion, or by adding the latex into the emulsifier solution phase before processing, or by comilling the latex with the bitumen, water and emulsifier all resulted in a network formation of bitumen particles surrounded by a continuous polymer film. The use of emulsified binders appears to result in a more evenly distributed polymer network compared to the use of hot polymer-modified binders, and they therefore have greater potential for consistent binder cohesion strength, stone retention and therefore improved pavement performance. PMID- 11903803 TI - Confocal laser scanning microscopy as a tool to determine cyanobacteria biomass in microbial mats. AB - We have developed a method based on confocal laser scanning microscopy for detection and quantification of cyanobacteria from the Ebro Delta microbial mats. Cyanobacteria play a major role as primary producers in microbial mats; it is difficult, however, to apply classical methods to estimate their biomass because they establish strong interactions with detritic particles. The protocol described here allows the localization of individual cells or filaments with micrometre precision without the need to either manipulate or stain the samples. This method is suitable for studying biomass 'in situ' in microbial mats. PMID- 11903806 TI - Neuroendocrinology with feeling. PMID- 11903807 TI - Targeted cytotoxic analogue of luteinizing hormone-releasing hormone (LH-RH) only transiently decreases the gene expression of pituitary receptors for LH-RH. AB - A cytotoxic analogue of LH-RH, AN-207, consisting of 2-pyrrolinodoxorubicin (AN 201) linked to carrier [D-Lys6]LH-RH, was developed for targeted therapy of cancers expressing LH-RH-receptors. To determine its possible side-effects on the pituitary gland, we investigated the gene expression of pituitary LH-RH-receptors and LH secretion in ovariectomized female and normal male rats after treatment with the maximum tolerated dose of AN-207. The effect of AN-207 on the gene expression of the pituitary GH-RH-receptors and GH secretion was also assessed in male rats. Five hours after a single i.v. injection of AN-207 at 175 nmol/kg, there was a 39-51% decrease in mRNA expression for the pituitary LH-RH-receptors in male and female rats. The carrier, at an equimolar dose, caused a similar reduction (37-39%), whereas the cytotoxic radical AN-201, at an equitoxic dose (110 nmol/kg), produced only a 12-24% decrease (NS) in the mRNA expression of LH RH-receptors. AN-207 and the carrier analogue induced a comparable 90-100-fold increase in serum LH concentrations in male rats, and the same 12-fold elevation in OVX rats at 5 h. Seven days after treatment with AN-207, the mRNA levels for the LH-RH receptors and the serum LH concentration were back to normal in both sexes. AN-207, the carrier, and AN-201 had no significant effect on the expression of mRNA for GH-RH-receptors in the pituitary. In vitro, a continuous perfusion of pituitary cells with 10 nM AN-207 did not affect the hormone releasing function of the targeted LH cells or the nontargeted GH cells. Our results demonstrate that cytotoxic LH-RH analogue AN-207, at the maximum tolerated dose causes only a transient decrease in the gene expression of the pituitary LH-RH receptors, and the levels of mRNA for LH-RH receptor fully recover within 7 days. Moreover, the carrier hormone moiety, and not the cytotoxic radical in AN-207 is responsible for this transient suppression. Our findings suggest that the therapy with cytotoxic LH-RH analogues will not inflict permanent damage to pituitary function. PMID- 11903808 TI - Nocturnal accumulation of cyclic 3',5'-guanosine monophosphate (cGMP) in the chick pineal organ is dependent on activation of guanylyl cyclase-B. AB - The role of cGMP in the avian pineal is not well understood. Although the light sensitive secretion of melatonin is a well-known output of the circadian oscillator, pharmacologically elevated cGMP levels do not result in altered melatonin secretory amplitude or phase. This suggests that pineal cGMP signalling does not couple the endogenous circadian oscillator to the expression of melatonin rhythms. Nonetheless, the free-running rhythm of cGMP signalling implies a link to the circadian oscillator in chick pinealocytes. As the circadian rhythm of cGMP levels in vitro is not altered by pharmacological inhibition of phosphodiesterase activity, we infer that the synthesis, rather than the degradation of cGMP, is under circadian control. In vitro experiments with the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine as well as with an inhibitor of the NO-sensitive soluble guanylyl cyclase (sGC), showed that the NOS-sGC pathway does not play a major role in the circadian control of cGMP generation. In organ culture experiments, we demonstrated that C-type natriuretic peptide (CNP), but not atrial natriuretic peptide (ANP), elevated daytime levels of cGMP. As CNP acts on the membrane guanylyl cyclase isoform B (GC-B), which is expressed at very high levels in mammalian pineals, we investigated the effect of the membrane GC-specific inhibitor HS-142 on chick pineal cGMP levels. CNP induced daytime cGMP levels were reduced by HS-142. More importantly, 'spontaneously' high nocturnal levels of cGMP in vitro were reduced to daytime levels by acute addition of HS-142. These data implicate endogenous nocturnal CNP release and subsequent activation of GC-B as the major input driving cGMP synthesis in the chick pineal organ. PMID- 11903809 TI - Distribution of androgen receptor mRNA expression and immunoreactivity in the brain of the green anole lizard. AB - Male courtship and copulation are androgen dependent in the green anole lizard, and female receptivity can be facilitated by testosterone. However, only a few, and relatively large, regions in the brain have been implicated in the control of these behaviours. In situ hybridization and immunohistochemistry were therefore used to determine in detail where androgens are likely to act in the brains of breeding males and females. A 697-bp fragment of the anole androgen receptor (AR) was cloned from total RNA isolated from the kidney, which contains the highly androgen-sensitive renal sex segment. The cloned fragment spanned part of the C, the entire D, and part of the E domains, and shared a high degree of similarity with the AR of various species. 35S-labelled antisense and sense probes were generated from the 697-bp fragment for use in in situ hybridization, and the AR antibody PG-21 was used for immunohistochemistry. Both sexes consistently had AR mRNA expression and immunoreactivity in areas associated with vertebrate reproductive behaviours and in motor areas of the brainstem. Interestingly, the PG-21 antibody produced labelling in both the nucleus and cytoplasm, including neuronal processes. The distribution of mRNA and immunoreactivity were comparable in males and females, and the amount of labelling was generally similar, although slightly greater in females. The expression pattern of AR in this species supports the idea that distribution is highly conserved among vertebrates, but that it probably does not dictate behavioural differences between the sexes in anoles. PMID- 11903810 TI - Sympathoadrenal system differentially affects photoperiodic changes in humoral immunity of Siberian hamsters (Phodopus sungorus). AB - Siberian hamsters (Phodopus sungorus) rely on photoperiod as a primary cue to coordinate seasonally appropriate changes in physiology and behaviour. Among these seasonal changes is reduced immune function in short 'winter-like' days, compared to long 'summer-like' days. Previous evidence suggests that immune function is regulated, in part, by the sympathoadrenal system. The precise role of the sympathoadrenal system in regulating photoperiodic changes in immune function, however, remains unspecified. The goal of the present study was to examine the differential contributions of direct sympathetic innervation of immune target tissue, as well as adrenal medullary catecholamines, to photoperiodic changes in immune function in male Siberian hamsters. In Experiment 1, hamsters underwent either bilateral surgical removal of the adrenal medulla (ADMEDx), or sham surgeries, and were maintained in long (LD 16 : 8) or short days (LD 8 : 16). In Experiment 2, hamsters received either surgical denervation of the spleen, or sham surgeries, and were then housed in long or short days. Serum anti-KLH IgG concentrations and splenic norepinephrine (NE) content were determined in both experiments. Short-day hamsters had reduced humoral immunity compared to long-day hamsters. ADMEDx reduced immune function, but only in long day hamsters. In contrast, splenic denervation reduced humoral immunity, but only in short-day hamsters. Splenic NE content was increased in short days and by ADMEDx. NE content was markedly reduced in denervated hamsters compared to sham operated hamsters. Collectively, these results suggest that the sympathoadrenal system is associated with photoperiodic changes in immune function. PMID- 11903813 TI - Centrally regulated blood pressure response to vasoactive peptides is modulated by corticosterone. AB - To investigate the role of brain glucocorticoid (GR) and mineralocorticoid receptors (MR) in centrally evoked blood pressure responses, the effects of intracerebroventricular (i.c.v.) administration of angiotensin II and vasopressin were studied in adrenalectomized rats with and without corticosterone or aldosterone replacement. Five groups were examined: (i) Adrenalectomy (ADX); (ii) ADX + a subcutaneously implanted 20-mg corticosterone pellet (low corticosterone); (iii) ADX + 100 mg corticosterone pellet (high corticosterone); (iv) ADX + 6 microg/24 h aldosterone via Alzet minipump (Aldo); and (v) Sham adrenalectomy (Sham). Pressor responses to 150 ng angiotensin II and 50 ng vasopressin i.c.v. were determined in freely moving rats using biotelemetry. The results show that, compared to sham rats, ADX rats showed significantly reduced pressor responses. This reduction of the pressor response to angiotensin II could be reversed and even further enhanced by replacement of the ADX rats with high corticosterone concentrations. In contrast, with aldosterone, a depressor type response was observed. Corticosterone replacement could not restore the pressor response to vasopressin. We conclude that the pressor response to centrally administered vasoactive substances is substantially attenuated by removal of the adrenals and that, in the case of angiotensin II, this is due to the lack of high concentrations of circulating corticosterone occupying both MR and GR. However, predominant MR occupancy appears to play an opposite role and attenuates the angiotensin II-induced pressor response. PMID- 11903811 TI - Noradrenaline and dopamine regulation of prolactin secretion in sheep: role in prolactin homeostasis but not photoperiodism. AB - The role of noradrenaline (NA) and dopamine (DA) in the hypothalamic control of prolactin (PRL) secretion was investigated in hypothalamic intact (control) and hypothalamo-pituitary disconnected (HPD) Soay rams. The animals were exposed to alternating 16-weekly periods of short (8 L : 16D) and long days (16 L : 8D) to induce marked cyclical changes in PRL secretion in both groups (as demonstrated previously). Selective NA and DA receptor antagonists (dose: 1.2 micromol/kg) were administered under short days (low endogenous PRL secretion), and agonists (dose: 0.0012-0.12 micromol/kg) were administered under long days (high endogenous PRL secretion). The acute changes in blood PRL concentrations were measured over 4 h as the index of responsiveness. Under short days, treatment with WB4101 (alpha-1 adenoceptor antagonist), and rauwolscine (alpha-2 antagonist), consistently increased PRL secretion in control, but not in HPD rams. The treatments produced similar acute, drug-specific behavioural effects in both groups. Propranolol (beta antagonist) had no effect on PRL secretion, while sulpiride (DA D-2 antagonist) induced a marked increase in blood PRL concentrations in control rams (> 4 h), and a transient effect in HPD rams (15 min). Under long days, when endogenous PRL secretion was increased, phenylephrine (alpha-1 agonist) produced no effects, while bromocriptine (DA D-2 agonist) robustly decreased PRL concentrations in both control and HPD rams, even at the lowest treatment dose. Overall, the positive responses to the antagonists in the control rams, support the view that DA (acting via D-2 receptors), and to a lesser extent NA (acting via alpha-1/alpha-2 receptors), negatively regulate PRL secretion. In contrast, the lack of responses to the antagonists in the HPD rams, support the view that neither DA, nor NA, mediate the photoperiodic control of PRL secretion. PMID- 11903812 TI - GABA mediates steroid-induced astrocyte differentiation in the neonatal rat hypothalamus. AB - Our previous work has demonstrated that astrocytes in the developing arcuate nucleus of the rat hypothalamus are sexually dimorphic as a result of differential exposure to oestradiol. Moreover, our experiments in neonatal rats suggest an absence of oestrogen receptors (ER) in arcuate nucleus astrocytes in vivo. This, along with the conspicuous lack of evidence in the literature confirming the presence of ER in arcuate nucleus astrocytes of the neonatal rat brain, led us to question the mechanism by which oestrogen induces changes in arcuate nucleus astrocyte morphology. Based on our previous findings that oestradiol increases gamma-aminobutyric acid (GABA) levels in the neonatal rat arcuate, we hypothesize that GABA is released from neighbouring oestrogen sensitive neurones and alters arcuate nucleus astrocyte morphology. Here, we report that in vivo reduction of GABA synthesis in the neonatal rat brain by antisense oligodeoxynucleotides to glutamic acid decarboxylase prevented gonadal steroid-induced astrocyte differentiation in males and testosterone-treated females. Conversely, activation of GABAA receptors with the agonist muscimol increased astrocyte differentiation in females in the absence of gonadal steroids. Given that GABA is made only in neurones and that its synthesis is increased by oestradiol, we conclude that it acts as a diffusible factor inducing the differentiation of neighbouring astrocytes. PMID- 11903814 TI - Involvement of postsynaptic EP4 and presynaptic EP3 receptors in actions of prostaglandin E2 in rat supraoptic neurones. AB - We have reported that supraoptic nucleus (SON) neurones are excited by prostaglandin E2 (PGE2) presumably via dual postsynaptic PG receptors, FP receptors and unidentified EP receptors, and that presynaptic EP receptors may also be involved in the excitation. In the present study, to clarify the receptor mechanism of the PGE2-mediated actions on SON neurones, we studied the pre- and postsynaptic effects of four newly developed EP agonists that are selective for each of the four EP receptors, EP1-4, on rat SON neurones using extracellular recording and whole-cell patch-clamp techniques. The EP4 agonist ONO-AE1-329 mimicked the excitatory effects of PGE2, whereas the EP1 agonist ONO-DI-004, the EP2 agonist ONO-AE1-257 and the EP3 agonist ONO-AE-248 had little or no effect. The effects of ONO-AE1-329 were unaffected by the EP1/FP/TP antagonist, ONO-NT 012, which potently suppressed the excitation caused by the FP agonist fluprostenol and PGE2. ONO-AE1-329 caused marked excitation when responses to fluprostenol were desensitized by repeated applications of fluprostenol. Patch clamp analysis in SON neurones showed that ONO-AE1-329 induced inward currents at a holding potential of -70 mV and the reversal potential of the currents was 35.1 +/- 2.3 mV. On the other hand, the frequency of spontaneous inhibitory postsynaptic currents recorded from SON slice preparations was suppressed by ONO AE-248, but unaffected by the other three EP agonists. These results suggest that SON neurones possess postsynaptic EP4 receptors and that gamma-aminobutyric acid neurones innervating SON neurones possess presynaptic EP3 receptors in their terminals. Activation of the two EP receptors may be involved in the excitatory regulation of SON neurones by PGE2. PMID- 11903815 TI - Rapid stimulation of the PI3-kinase/Akt signalling pathway in developing midbrain neurones by oestrogen. AB - Oestrogen promotes the differentiation of neurones in the central nervous system. In the rodent midbrain, the maturation of dopaminergic neurones appears to be under oestrogen control. This is supported by the fact that dopaminergic cells contain nuclear oestrogen receptors-alpha/beta (ER). Second, aromatase is transiently expressed in the developing midbrain. In previous studies, we have shown that oestrogen increases dopamine synthesis and plasticity of dopamine cells. These effects are transmitted through classical nuclear ER but require also the stimulation of nonclassical signalling pathways involving the activation of membrane receptors. This study attempted to identify nonclassical oestrogen dependent signalling cascades which might be stimulated downstream of membrane ERs. Using cultured mouse midbrain cells, we could demonstrate by Western blotting, that oestrogen rapidly phosphorylates Akt, a kinase which is implicated in the phosphatidylinositol 3 (PI3)-kinase pathway. This effect was only seen in midbrain neurones but not astrocytes. Oestrogen-induced Akt phosphorylation was time- and dose-dependent, showing highest responses after 30 min and at a steroid concentration of 10(-8) and 10(-6) M. Immunocytochemistry for phosphorylated Akt (pAkt) demonstrated that pAkt is predominantly found in a nuclear/perinuclear position and that oestrogen exposure increased the number of pAkt-positive cells. To investigate the mechanisms which are involved in transmitting oestrogen effects on the cellular level, cells were treated with antagonists for distinct signalling pathways. The application of the nuclear ER antagonist ICI 182 780 did not abolish the oestrogen-induced Akt phosphorylation. In contrast, interrupting intracellular calcium signalling with BAPTA completely prevented this effect. The PI3-kinase inhibitor LY294002 also inhibited the activation of Akt by oestrogen. Our study clearly indicates that oestrogen can rapidly stimulate the PI3 kinase/Akt signalling cascade in differentiating midbrain neurones. This effect requires the intermediate activation of calcium-dependent signalling pathways. In conclusion, oestrogen effects in the developing midbrain appear to be connected with the PI3-kinase/Akt signalling mechanism. PMID- 11903816 TI - Ghrelin: a newly discovered hormone. AB - Ghrelin is a gastric peptide that stimulates growth hormone secretion and increases adiposity. It is the first identified natural ligand for a previously cloned growth hormone secretagogue receptor which is present in the pituitary gland and the hypothalamic region of the brain. Insights into its regulation will now enable its physiological role to be elucidated and its potential clinical use evaluated. PMID- 11903817 TI - The unexplained survival of cells in oral cancer: what is the role of p53? AB - In normal oral epithelium the cells divide, mature, differentiate, and die. This sequence is not normally followed in oral cancer. Instead, the death of the cells is somehow prevented, although the pathways toward cell death in normal oral epithelium and the defects in oral cancer are not well defined. However, several components in the system have been identified, and information on their interactions is becoming available. This review summarizes the evidence for cell death being due to apoptosis and the central role of the p53 gene product in its regulation. Areas for future research are also identified. PMID- 11903818 TI - Comparison of HPV infection, p53 mutation and allelic losses in post-transplant and non-posttransplant oral squamous cell carcinomas. AB - BACKGROUND: Oral squamous cell carcinoma (SCC) is increasingly found in transplant recipients, although little is known of the natural history of the disease or the mechanism underlying this increase. METHODS: In this article we describe the history of development of 5 oral post-transplant SCCs (PSCCs) and compare their genetic profiles to 34 non-posttransplant SCCs (NPSCCs). RESULTS: Of the five patients with PSCCs, 3 had bone marrow transplants and two, kidney. All three PSCCs from bone marrow recipients were preceded locally by graft-vs. host disease (GVHD). Two of the GVHD were biopsied and demonstrated dysplasia. Similar frequencies of loss of heterozygosity (LOH) occurred in PSCCs and NPSCCs at 3p, 9p, 17p and 8p, with lower frequencies in PSCCs at 4q (39% vs. 0%), 11q (53% vs. 20%) and 13q (45% vs. 20%), although the latter were not significantly different. Only 1 PSCC had a p53 mutation, compared to historical values of 40 60% for NPSCC. Interestingly, human papillomavirus (HPV) DNA was detected in 3 (60%) PSCCs, in comparison to only 4 (12%) of the 34 NPSCCs (P = 0.0346). CONCLUSIONS: Dysplasia in oral GVHD may be a strong indicator of cancer risk and should not be regarded as reactive changes to lichenoid mucosites. The low level of p53 mutation and increased HPV infection support the involvement of HPV in the development of PSCC, while the similarity in LOH patterns suggests that other aspects of carcinogenesis may be comparable in these two types of SCCs. PMID- 11903819 TI - Altered cytokeratin expression during chemoprevention of experimental hamster buccal pouch carcinogenesis by garlic. AB - BACKGROUND: Cytokeratins (also known as keratins (K)) are members of the family of intermediate filaments and form major components of the mammalian epithelial cell cytoskeleton. Cytokeratins have emerged as reliable cellular markers of oral cancer development and chemoprevention because of their abundance, stability and high antigenicity. METHODS: We investigated the effect of aqueous garlic extract on cytokeratin expression during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. Hamsters were divided into four groups of six animals. Animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin, on the right buccal pouches, three times a week for 14 weeks. Group 2 animals were painted with DMBA as in group 1 and also received 250 mg/kg body weight aqueous garlic extract orally on alternate days to the DMBA application. Group 3 animals received garlic extract only, as in group 2. Group 4 animals received neither DMBA nor garlic extract and served as the control. The hamsters were killed after an experimental period of 14 weeks. RESULTS: Cytokeratin expression was studied using human monoclonal antibodies AE1 and AE3, which react with type I and II keratins. In DMBA-induced squamous cell carcinomas, decreased expression of high molecular weight keratins was observed. Administration of garlic extract to animals painted with DMBA suppressed HBP carcinomas and restored normal cytokeratin expression. CONCLUSION: The results of the present study suggest that inhibition of HBP carcinogenesis by garlic may be due to its regulatory effects on differentiation, tumour invasiveness, migratory and metastatic potential. We suggest that one of the mechanisms of tumour inhibition by garlic is an influence on cellular differentiation. PMID- 11903820 TI - Rising trends in oral cancer mortality in Spain, 1975-94. AB - BACKGROUND: Epidemiological studies have shown that the worldwide incidence and mortality rates for oral cancer have increased considerably over the last decade. This study investigates the Spanish trend in mortality of oral cancer from 1975 94. METHODS: Age-standardized, truncated, cumulative, age-specific and potential years of life lost (PYLL) rates were calculated by gender. Poisson regression models allowed the measurement of age and period effects. RESULTS: Age standardized and cumulative mortality rates increased in males, while truncated and PYLL rates doubled. Changes were less marked in females. There were annual increases in oral cancer mortality from 1975-94, of 25% and 9% in males and females, respectively. In males there was an interaction between age and period. There was also an increase in age-specific mortality rates in males. CONCLUSIONS: Mortality from oral cancer rose substantially in males, with concomitant changes in the age-distribution pattern of mortality. Increases were less marked in females, with some minor modifications to the age-distribution pattern of mortality. PMID- 11903822 TI - Expression of RANTES and CCR1 in oral lichen planus and association with mast cell migration. AB - BACKGROUND: T lymphocytes and mast cells infiltrate the lamina propria in oral lichen planus (OLP). Chemokines and their receptors are involved in T cell and mast cell migration and accumulation during the inflammatory process. METHODS: In the present study, we investigated the role of RANTES and its receptors in OLP using immunohistochemistry, RT-PCR and an in vitro chemotaxis assay. RESULTS: RANTES and CCR1 were expressed on T cells and mast cells in OLP, while OLP lesional T cell supernatants stimulated CCR1 mRNA expression in a human leukemia mast cell line (HMC-1). TNF-alpha stimulated CCR1, CCR4 and CCR5 mRNA expression in the same cell line. OLP lesional T cell supernatants stimulated HMC-1 migration, which was partly inhibited by anti-RANTES antibody. CONCLUSIONS: The present study shows, for the first time, the distribution of RANTES and CCR1 in OLP. It is hypothesized that RANTES and CCR1 may play important roles in mast cell trafficking and related events in OLP. PMID- 11903821 TI - Quantification of oral mucositis due to radiotherapy by determining viability and maturation of epithelial cells. AB - BACKGROUND: An in-vitro assay has been developed for quantitative assessment of chemotherapy induced oral mucositis. In the present study this method was evaluated for assessment of irradiation mucositis at a cellular level. METHODS: Ten patients participated in this consecutive study. All patients were treated with conventional fractionated curative postoperative radiotherapy. Prior to, and weekly during, the irradiation course, oral washings were obtained to determine viability of epithelial cells by trypan blue dye exclusion. Maturation of epithelial cells was assessed from smears (Papanicolaou staining). The viability data were compared with the WHO-score for mucositis. RESULTS: Epithelial cell viability increased during the first three weeks of radiation (P = 0.04), and was seen earlier than the subjective mucosal changes with the WHO-score. Cell maturity shifted from immature and intermediate to mature (P = 0.03). CONCLUSIONS: The cell viability assay can be considered an objective method for following the development of irradiation mucositis, and seems to be more sensitive during the first three weeks of irradiation than the WHO-scoring method. PMID- 11903823 TI - Co-existence between oral lesions and opportunistic systemic diseases among HIV infected subjects in Thailand. AB - BACKGROUND: The purpose of this study was to determine whether any relationship exists between the occurrence of oral lesions and opportunistic systemic diseases among HIV-infected subjects. METHODS: A cross-sectional analytical study was performed in two hundred and seventy-eight HIV-infected heterosexual persons and intravenous drug users (IVDUs)(230 males and 48 females, aged 16-65 years, mean 31.9 years). Eighty-six HIV-free subjects from the same population were included as controls (61 males and 25 females, aged 17-63 years, mean age 33.1 years). The following information was recorded for each patient: age, gender, risk group and stage of HIV infection, immune status, medication, systemic disease and presence of oral lesions. RESULTS: Oral candidiasis was the most common oral lesion among HIV-infected individuals (40%), followed by hairy leukoplakia (HL)(26%). The three most common systemic diseases among the subjects were tuberculosis (TB)(53%), cryptococcosis (14%) and Pneumocystis carinii pneumonia (PCP)(11%). Logistic regression analysis revealed a significant association between the occurrence of TB and the presence of oral candidiasis (OR 2.8; 95% CI 1.6-4.8; P < 0.001), and the occurrence of PCP and the presence of HL (OR 2.2; 95% CI 1.1 4.3; P < 0.001). Positive predictive values of any oral lesions and oral candidiasis in predicting TB were 87% (95% CI 73.0-94.6) and 67% (95% CI 51.9 80.0), respectively. CONCLUSIONS: We concluded that oral candidiasis might be used as a clinical marker for TB, and HL for PCP. Recognition of the lesions by health-care providers may indicate the need for more intensive clinical and laboratory monitoring and possibly initiation of prophylaxis against these opportunistic systemic infections. PMID- 11903824 TI - Oral manifestations of an HIV positive cohort in the era of highly active anti retroviral therapy (HAART) in South London. AB - BACKGROUND: Human Immunodeficiency Virus (HIV) infection is associated with oral manifestations of diagnostic and prognostic importance. With the advent of Highly Active Anti-retroviral Therapy (HAART) there is anecdotal evidence to suggest that the prevalence of oral lesions has declined. The number of prevalence studies, carried out in the era of HAART is, however, meagre. Our aim was to study the prevalence of the oral manifestations of HIV in a population, predominantly on HAART, attending a Genito-Urinary Medicine Centre in South London. METHODS: This cross sectional study included 203 adult volunteers, comprising 76% males and 24% females. One third of the subjects were from the predominantly African or Afro- Caribbean ethnic minority groups resident in London. The relationship between the prevalence of oral lesions and demographic variables, therapeutic regimes, viral load and CD4 counts were evaluated. RESULTS: One hundred (49%) of the patients had no detectable oral lesions. Oral lesions detected most frequently included oral hairy leukoplakia (9.9%), HIV associated periodontal diseases (9.9%) and oral candidiasis (4.9%). Three subjects had multiple papillomatous growths. Most cases (n = 17/20) of oral hairy leukoplakia were in individuals with a detectable (> 400 copies/ml) plasma RNA viral load. The majority (n = 8/10) of our patients with oral candidiasis had a plasma RNA viral load > 10,000 copies/ml and half (n = 5/10) had a CD4 count < 200 cells/mm3. Logistic regression analysis suggested that the presence of an oral lesion was not associated with any demographic features except for periodontal diseases which were associated with tobacco smoking (P = 0.023). CONCLUSIONS: The prevalence of so called 'strongly associated' oral lesions of HIV is low in this South London HIV-infected population on HAART, and the relative frequency is different from that cited in the literature from the pre HAART era. The oral lesions detected were found mostly in people with low CD4 counts and high HIV-1 RNA viral loads, suggesting they were very immunocompromised, not on, or declining therapy, or that their therapy was failing. PMID- 11903825 TI - Involvement of substance P, mast cells, TNF-alpha and ICAM-1 in the infiltration of inflammatory cells in human periapical granulomas. AB - BACKGROUND: Substance P (SP), a potent proinflammatory peptide present in sensory neurons, is believed to be a major mediator of neurogenic inflammation. The aim of this study was to examine the localization and involvement of SP, mast cells and tumor necrosis factor-alpha (TNF-alpha)-positive cells in human periapical granulomas. METHODS: Sections from seven periapical granulomas were stained using a variety of immunohistochemical methods. RESULTS: SP-immunoreactive nerve fibers, mast cells and TNF-alpha-positive cells were found localized in the vicinity of blood vessels in all the samples of periapical granulomas. The vascular endothelial cells stained positively for E-selectin and intercellular adhesion molecule-1. SP, TNF-alpha-positive cells and E-selectin could not be detected in clinically healthy periodontal ligament, and served as a negative control. CONCLUSION: Our findings suggest that SP, mast cells, TNF-alpha-positive cells and E-selectin may modulate the pathogenesis of apical periodontitis and may be responsible for stimulating the formation of granuloma with the resorption of periapical bone. PMID- 11903826 TI - Clear cell odontogenic carcinoma with ghost cells and inductive dentin formation report of a case in the mandible. AB - An atypical case of clear cell odontogenic carcinoma occurred in the mandible of a 50-year-old Japanese man. Microscopically, the tumor mainly consisted of clear, glycogen-rich epithelial cell nests, aggressively proliferated in the mandible and invading into the surrounding soft tissues. Within the tumor cell nests, clusters of ghost cells were observed in addition to occasional foci of microcystic degeneration. Hyaline, osteoid or dentinoid-like deposits were also demonstrated adjacent to the tumor cell nests. Dentinal tubules were clearly revealed in some of these structures. Hence, this tumor was regarded as an epithelial neoplasm with odontogenic ectomesenchyme induction. PMID- 11903827 TI - Uncommon dermoid cyst presented in the mandible possibly originating from embryonic epithelial remnants. AB - A case of an intraosseous dermoid cyst that had developed in the mandible of a 29 year-old-male is reported. The patient was admitted to our dental clinic complaining of pain in the right molar area of the mandible. The patient had no particular history of the present illness. A radiographic examination revealed diffuse bone absorption in the right molar area. A biopsy showed the lesion to be an intraosseous dermoid cyst which was surgically enucleated. Microscopic examination of the excised tissue showed it to be well circumscribed with fibrous tissues, and the cystic space lined by keratinized epithelium. In addition, sweat glands and hair follicle-like skin appendages were identified in the cystic wall, where a daughter cyst was also found. We suggested that this uncommon cyst had developed in the mandible, as a result of embryonic epithelium migration. The patient remained in good health during the three year follow-up. PMID- 11903829 TI - Should parents assess the interpersonal skills of doctors who treat their children? A literature review. AB - Medical consultations in which doctors display good interpersonal skills are associated with a wide range of positive health outcomes. Obtaining the perceptions of patients, or parents in paediatric settings, regarding the interpersonal skills demonstrated by their doctors could provide feedback on doctor behaviours that influence health outcomes. It could also offer an alternative to more traditional methods of assessing interpersonal skills, such as using standardized patients. Patient perceptions of doctor interpersonal skills are most commonly obtained through patient-completed satisfaction questionnaires. A literature review was conducted with the aim of examining the potential role of parent perceptions in the evaluation of paediatric interviews. Studies identified were reviewed for information regarding the rationale for obtaining parent perceptions, the reliability and validity of measures used, the association between parent evaluations and specific doctor interview behaviours and the acceptability and feasibility of obtaining this information. Practical applications of the information obtained from parent evaluations were also sought. There was considerable support for the inclusion of patient evaluations in the assessment of the interpersonal skills of medical students and doctors. Reliable and valid measurement of patient evaluations can be obtained and patients are willing to provide this information. A clear association between specific doctor interview behaviours and parent satisfaction ratings was demonstrated. However, the important issue of feasibility and acceptability to doctors and medical students of obtaining patient perceptions of interpersonal skills, was not addressed. Patient evaluations of the interpersonal skills displayed by their doctors should be a component of clinical skills assessments. Information obtained from parents relating to the care of children could provide feedback to doctors regarding their personal interviewing style. In order to make full use of the information obtained from parents, there is a need for further study to establish the sensitivity of parent evaluations and methods to facilitate the process of obtaining this information. PMID- 11903828 TI - Perspectives on infective ear disease in indigenous Australian children. AB - Australian Aboriginal children experience the highest rates of bacterial respiratory diseases reported in the literature. Neonatal acquisition of multiple bacterial pathogenic species and strains predicts persistent and severe disease throughout childhood, particularly infective ear disease. The dynamics of bacterial nasopharyngeal colonization and transmission are poorly understood. The importance of host factors, bacterial competition and co-operation in the transition from asymptomatic carriage to disease are also uncertain. Treatment outcomes are poor, possibly due to the high density of bacterial infection following early age exposure, poor compliance and increasing levels of antibiotic resistance. The relationship between antibiotic use, clinical outcomes and bacterial resistance needs to be better understood in high-risk populations if the benefits associated with treatment are to be maximized. While there is an urgent need for vaccines, the early age of infection and the high rates of transmission and bacterial antigenic diversity mean these may also be less effective than predicted from studies in low-risk populations. PMID- 11903831 TI - Shared bathing and drowning in infants and young children. AB - OBJECTIVE: A study was undertaken to look at possible risks of shared bathing in early childhood. METHODS: Autopsy databases were searched at the Women's and Children's Hospital and State Coroner's Office, Adelaide, Australia, from January 1963 to December 1999, the Victorian Institute of Forensic Medicine, Melbourne, Australia, from January 1991 to December 1999, and the Children's Hospital-San Diego, San Diego, USA, from January 1990 to December 1999, for all cases registered as drowning in children aged 2 years and under who were in a bath with another child around the time of death. RESULTS: A total of 17 cases were found. The age range of the victims was 8-22 months (average=11.8 months), with a male to female ratio of 10:7. The surviving children (who were all siblings) were significantly older, with an age range of 19-48 months (average=30.4 months), and a male to female ratio of 12:5. (The survivors were on average 18.5 months older than the victims, range=11-32 months). In every case the children had been left unsupervised for variable periods of time. CONCLUSIONS: These cases represented a significant component -- between 22 and 58% -- of bath drownings in the 2 years and under age group. It would appear that shared bathing for young children and infants may only acceptable if adult supervision is constant, and that a young sibling is not an appropriate person to look after an infant or toddler in the bath. While the risk of leaving an infant in a filled bath may be exacerbated if an older child is also present, further population-based studies are required to examine this hypothesis. PMID- 11903830 TI - Discharge guidelines for children with acute asthma: a consensus statement. PMID- 11903832 TI - Comparison of Dinamap 8100 with sphygmomanometer blood pressure measurement in a prepubertal diabetes cohort. AB - OBJECTIVE: To compare the conventional sphygmomanometer with the semiautomated Dinamap 8100 (Critikon, Tampa, FL, USA) for the measurement of blood pressure in prepubertal children with insulin-dependent diabetes mellitus. METHODOLOGY: Blood pressure was measured using both methods in 61 prepubertal children (aged 8-13 years) on 189 occasions over 4 years. The measurements were compared using the Bland-Altman plot. Tracking correlations of blood pressure centiles over time were analyzed by the general estimating equation. RESULTS: Accuracy criteria of the Association for the Advancement of Medical Instrumentation were met and a British Hypertensive Society 'B' grading was reached. Differences in systolic and diastolic blood pressure were found between the two methods (P < 0.01). For systolic blood pressure, common correlations were 0.54 (Dinamap) and 0.51 (sphygmomanometer) and for diastolic blood pressure were 0.33 and 0.42, respectively. CONCLUSION: The Dinamap 8100 is an acceptable alternative in clinic practice and research for prepubertal children. PMID- 11903833 TI - Low-density lipoprotein subclasses in children under 10 years of age. AB - OBJECTIVE: To examine the prevalence of small dense low density lipoprotein (LDL) in a cohort of children under 10 years of age and to investigate the relationship to other biochemical variables and to measures of fatness. A preponderance of small dense LDL (pattern B), is associated with obesity, abdominal fat accumulation, insulin resistance and risk of heart disease in adults. METHODOLOGY: LDL peak particle diameter (PPD) was determined by gel electrophoresis in 53 children under 10 years of age and in 65 of their parents: apoproteins A1 and B were determined by turbidimetry. Anthropometric variables, basic lipid profiles, insulin and leptin had been determined previously. Differences between patterns A (large light particles > 25.5 nm diameter) and B were examined by t-test, Chi-square, or Mann-Whitney test. Relationships between the variables were reported as Pearson correlation coefficients. RESULTS: Pattern B (PPD of < or = 25.5 nm) prevalence was 7.5% in children and 11% in parents (17% in men and 5% in women). Most of the children (86%) who had PPD < or = 26.0 nm also had parents with PPD in this range. A strong association was found between children's and mother's PPD (r=0.60, P < 0.001), but this was somewhat less with fathers (r=0.40, P=0.02). Children in the lowest tertile of PPD had a tendency towards a higher body mass index, waist, fat mass and insulin. CONCLUSIONS: The prevalence of small dense LDL is lower in children under 10 years of age than in their parents; fathers had a higher prevalence of pattern B than mothers and there is some evidence of a familial effect in the inheritance of pattern B. PMID- 11903834 TI - Chest physiotherapy and porencephalic brain lesions in very preterm infants. AB - OBJECTIVE: National Women's Hospital is one of two hospitals to report a destructive brain lesion, namely encephaloclastic porencephaly (ECPE), in extremely preterm infants. It has been associated with non-cephalic presentation, early hypotension and the number of chest physiotherapy treatments in the first month. The aim of the present study was to determine the temporal relationship between ECPE and chest physiotherapy use in very low-birth weight (VLBW) infants in our unit. METHODOLOGY: Cerebral ultrasound scan reports, post-mortem reports, clinical and physiotherapy records and, if indicated, original ultrasound films were reviewed for all VLBW babies admitted between 1985 and 1998. RESULTS: Over the 14 year period in question, 2219 babies with a birth weight < or = 1500 g were admitted. Encephaloclastic porencephaly was found in only the 13 previously reported babies born between 1992 and 1994. Encephaloclastic porencephaly was excluded in 1564 (70%) babies. In 621 (28%) babies who did not have late ultrasound scans, ECPE was thought to be unlikely either because the babies never had any chest physiotherapy (n=479) or because they had chest physiotherapy but were known to be neurodevelopmentally normal on follow up (n=142). Data were incomplete for 21 babies (0.9%). The number of chest physiotherapy treatments per baby decreased from a median of 95 prior to 1989 to 38 and the age of starting treatment increased from 5 to 8 days after 1990. The use of chest physiotherapy ceased in 1995. CONCLUSIONS: Encephaloclastic porencephaly emerged as a problem at a time when the use of chest physiotherapy had decreased. The cluster of cases seen between 1992 and 1994, although associated with the number of chest physiotherapy treatments given, began to appear because of some other factor. PMID- 11903835 TI - Effect of cisapride on gastric emptying in premature infants with feed intolerance. AB - OBJECTIVE: To assess the effect of cisapride on gastric emptying and gastro oesophageal reflux (GOR) symptoms in preterm infants with feed intolerance. METHODS: Sixteen preterm infants (gestational age 24-35 weeks) with feed intolerance were enrolled in the study. Infants were randomized to receive 7 days of cisapride 0.2 mg/kg four times a day, immediately followed by 7 days of placebo or vice versa. Gastric emptying was measured using the [13C]-octanoic acid breath test prior to study entry and repeated on day 5, 6 or 7 after randomization and 5, 6 or 7 days after crossover. The symptoms of GOR were monitored during the study period using a standardized reflux chart. Weight was recorded daily. RESULTS: There was no change in gastric emptying in infants prescribed cisapride (gastric half-emptying time (t1/2) 31.9 +/- 4.7 vs 34.2 +/- 3.9 min for placebo vs cisapride, respectively; P = 0.65). Infants on cisapride had slower growth and there was no change in reflux symptoms. CONCLUSIONS: The use of cisapride in preterm infants with feed intolerance cannot be recommended. PMID- 11903836 TI - Seroprevalence of hepatitis B, hepatitis C and human immunodeficiency virus antibodies among multitransfused thalassaemic children in Shiraz, Iran. AB - OBJECTIVE: To evaluate the rate of seropositivity to hepatitis B and C and human immunodeficiency virus (HIV) infections among children with beta-thalassaemia major receiving multiple transfusions in Shiraz, Iran, compared with healthy controls. METHODOLOGY: The study was performed during 1999-2000 on multitransfused children with beta-thalassaemia major registered by the Shiraz Thalassaemia Society. Hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (HCV) antibodies (Ab) and HIV Ab were checked using a second-generation enzyme linked immunosorbent assay (ELISA). Positive tests were confirmed by western blots. Healthy blood donors were used for the control group. RESULTS: Hepatitis B surface antigen, anti-HCV Ab and HIV Ab were positive in four of 755 (0.53%, 95% confidence interval (CI)=0.17-1.3), 73 of 466 (15.7%; 95% CI=12.6-19.2) and none of 466 patients tested, respectively. Positive sera for HBsAg, anti-HCV Ab and HIV Ab were found in 85 (1.07%), 47 (0.59%) and 27 (0.34%) of 7879 control children, respectively. The rate of anti-HCV Ab was significantly higher in patients than in the control group (P < 0.0001). In patients, the rate of positive anti-HCV Ab was significantly higher than the rate for positive HBsAg (P < 0.0001). CONCLUSION: It is concluded that HCV is the current major problem in multitransfused children with thalassaemia major and more careful pretransfusion screening of blood for anti-HCV must be introduced in our blood banks. PMID- 11903837 TI - Scooter injuries in children. AB - OBJECTIVES: To study the causes, patterns of injury and use of safety equipment in children presenting with 'push/kick' scooter-related injuries. To draw comparisons with in-line skate, skateboard and bicycle injuries and to suggest strategies for injury prevention. METHODS: A retrospective review of medical data was undertaken for 12 consecutive months to September 2000. All children aged < 15 years who had attended the Sydney Children's Hospital with scooter, in-line skate (rollerblade), skateboard or bicycle injuries were identified. Children with scooter injuries for the latter 6 month period were contacted by telephone and interviewed, together with their parents, using a structured questionnaire. RESULTS: There was a marked rise in the number of scooter injuries from October 1999 to September 2000. Sixty-one per cent of these injuries occurred during the final 3 months of the study period, making scooters the most common cause of injury in the studied groups for this period. Forty-two per cent of scooter injuries were fractures. Only 3% of children used safety equipment at the time of injury, despite 86% owning some form of safety equipment. Children were less likely to use safety equipment with a scooter than with any other form of activity studied. (Chi-squared P=0.000). CONCLUSIONS: Scooters are a common cause of childhood injury, resulting in injury patterns similar to those caused by in line skates, skateboards and bicycles. Safety equipment is rarely worn when scooters are ridden. Injury patterns and riding styles suggest that if existing guidelines for in-line skating, skateboarding and bicycling are modified and applied to scooters, a reduction in injury numbers may be achieved. PMID- 11903838 TI - The epidemiology of invasive pneumococcal disease in children in Far North Queensland. AB - OBJECTIVES: To describe the epidemiology of invasive pneumococcal disease in children under 5 years of age in Far North Queensland and to examine the potential impact of a seven- and 11-valent conjugate pneumococcal vaccine. METHODS: A review of all cases of invasive pneumococcal disease in children under 5 years of age in Far North Queensland over a 9 year period (1992-2000). The distribution of the serotypes of isolates causing invasive pneumococcal disease was compared with the serotypes contained in the two vaccines. RESULTS: The annual incidence in indigenous and non-indigenous children under 5 years of age was 163 (95% confidence interval (CI) 122-213) and 42 (95% CI 31-55) cases per 100 000 children, respectively. For children under 2 years of age, these figures were 297 (95% CI 208-411) and 71 (95% CI 49-100), respectively. There was a greater variety of serotypes isolated from indigenous children (n=17) than from non-indigenous children (n=9; P < 0.01). The serotypes within the seven-valent vaccine accounted for 62% (95% CI 46-75%) and 88% (95% CI 76-95%) of the isolates from indigenous and non-indigenous children, respectively (P < 0.01). Serotypes within the 11-valent vaccine accounted for 72% (95% CI 57-84%) of the isolates from indigenous children under 5 years of age, but did not account for any extra isolates from non-indigenous children. CONCLUSION: Although the seven- and 11 valent conjugate pneumococcal vaccines cover only approximately 60 and 70%, respectively, of the isolates that cause invasive disease in indigenous children in Far North Queensland, they nevertheless have the potential to prevent much morbidity in and hospitalization of these children. It will be essential to maintain surveillance following the introduction of conjugate pneumococcal vaccines so as to monitor changes in the incidence of invasive pneumococcal disease, particularly in high-risk children. PMID- 11903839 TI - Maternal transmission of infectious pathogens in breast milk. AB - Breast milk provides a superior source of nutrition and many other health benefits to the newborn infant. However, breastfeeding has also been recognized as a route of transmission to the newborn of some important pathogens that may result in disease. In the present article, up-to-date research and recommendations on the maternal transmission of infectious pathogens in breast milk or by breastfeeding are discussed in the context of local epidemiology and in the setting of an industrialized country. PMID- 11903840 TI - Comparison of means and proportions using confidence intervals. PMID- 11903841 TI - An unusual cause of hepatitis. PMID- 11903842 TI - Cytomegalovirus infection in pregnancy. PMID- 11903843 TI - Pyridoxine-dependent seizures: a case report and a critical review of the literature. AB - Pyridoxine-dependent seizures are a recognized, although rare, cause of intractable seizures in neonates. Patients with this autosomal recessive disorder have recurrent seizures that are resistant to conventional anticonvulsants but respond dramatically to intravenous administration of pyridoxine. Life-long supplementation with pyridoxine is required to prevent seizure recurrence. In the absence of a biological marker for the disease, clinical diagnosis is often delayed and severe neurological sequelae are common. Herein, we report on the clinical course of a neonate with pyridoxine-dependent seizures. Delayed normalization of the electroencephalogram and a normal developmental outcome (at 15 months of age) on a dose of 10 mg/kg pyridoxine are distinctive features of the present case. We also review recent clinical observations and neurochemical studies that have added to our knowledge of this disorder. PMID- 11903844 TI - Cerebral vasculitis in a child following post-streptococcal glomerulonephritis. AB - Cerebral vasculitis following acute post-streptococcal glomerulonephritis (APSGN) is a rare neurological complication. An 11-year-old girl with biopsy proven APSGN developed an acute seizure disorder. Clinical and computed tomography findings were consistent with vasculitis. PMID- 11903845 TI - Asymptomatic bladder phaeochromocytoma in a 7-year-old boy. AB - Bladder phaeochromocytoma in a child is very rare. In the case presented here, it was an incidental finding in the youngest child reported in the literature. The phaeochromocytoma was located at the right ureterovesical junction causing obstruction, resulting in hydronephrosis. The dilemma in the diagnosis and management of this challenging condition is presented. The absence of typical symptoms of phaeochromocytoma is emphasized. PMID- 11903846 TI - Short falls and intracranial injury. PMID- 11903855 TI - Autonomic activity during human sleep as a function of time and sleep stage. AB - While there is a developing understanding of the influence of sleep on cardiovascular autonomic activity in humans, there remain unresolved issues. In particular, the effect of time within the sleep period, independent of sleep stage, has not been investigated. Further, the influence of sleep on central sympathetic nervous system (SNS) activity is uncertain because results using the major method applicable to humans, the low frequency (LF) component of heart rate variability (HRV), have been contradictory, and because the method itself is open to criticism. Sleep and cardiac activity were measured in 14 young healthy subjects on three nights. Data was analysed in 2-min epochs. All epochs meeting specified criteria were identified, beginning 2 h before, until 7 h after, sleep onset. Epoch values were allocated to 30-min bins and during sleep were also classified into stage 2, slow wave sleep (SWS) and rapid eye movement (REM) sleep. The measures of cardiac activity were heart rate (HR), blood pressure (BP), high frequency (HF) and LF components of HRV and pre-ejection period (PEP). During non-rapid eye movement (NREM) sleep autonomic balance shifted from sympathetic to parasympathetic dominance, although this appeared to be more because of a shift in parasympathetic nervous system (PNS) activity. Autonomic balance during REM was in general similar to wakefulness. For BP and the HF and LF components the change occurred abruptly at sleep onset and was then constant over time within each stage of sleep, indicating that any change in autonomic balance over the sleep period is a consequence of the changing distribution of sleep stages. Two variables, HR and PEP, did show time effects reflecting a circadian influence over HR and perhaps time asleep affecting PEP. While both the LF component and PEP showed changes consistent with reduced sympathetic tone during sleep, their pattern of change over time differed. PMID- 11903857 TI - Active processing of declarative knowledge during REM-sleep dreaming. AB - The ability to process recently acquired knowledge is clearly maintained during sleep. Here we assess whether and how far the sleeper controls this processing (in a non-volitional and non-conscious manner). We posit that during sleep, the cognitive concerns of previous waking may guide access to, and processing of, items of declarative knowledge with which those concerns are associated. In a delayed recall task, before each of three sleep onsets in the same night, 12 subjects heard a different nonsense sentence. When awakened in rapid eye movement (REM) sleep, they were asked to report their dream experience and to recall the last sentence heard. Occurrences of incorporation into dream content were more frequent for this sentence than for the sentences heard before previous sleep onsets, and also more frequent than occurrences of similar contents in reports from a control night. However, the modalities of elaboration of dream contents did not vary. These findings indicate that cognitive concern can affect the accessing of recently acquired declarative knowledge during sleep, but not the modalities by which this is inserted into dream content. They also suggest that cognitive concern may help consolidate knowledge by increasing the likelihood of it being processed during sleep. PMID- 11903856 TI - Slow release caffeine and prolonged (64-h) continuous wakefulness: effects on vigilance and cognitive performance. AB - Some long work or shift work schedules necessitate an elevated and prolonged level of vigilance and performance but often result in sleep deprivation (SD), fatigue and sleepiness, which may impair efficiency. This study investigated the effects of a slow-release caffeine [(SRC) at the daily dose of 600 mg] on vigilance and cognitive performance during a 64 h continuous wakefulness period. Sixteen healthy males volunteered for this double-blind, randomised, placebo controlled, two-way crossover study. A total of 300-mg SRC or placebo (PBO) was given twice a day at 21:00 and 9:00 h during the SD period. Vigilance was objectively assessed with continuous electroencephalogram (EEG), the multiple sleep latency tests (MSLT) and wrist actigraphy. Cognitive functions (information processing and working memory), selective and divided attention were determined with computerised tests from the AGARD-NATO STRES Battery (Standardised Tests for Research with Environmental Stressors). Attention was also assessed with a symbol cancellation task and a Stroop's test; alertness was appreciated from visual analogue scales (VAS). Tests were performed at the hypo (02:00-04:00 h, 14:00 16:00 h) and hypervigilance (10:00-12:00 h, 22:00-00:00 h) periods during SD. Central temperature was continuously measured and safety of treatment was assessed from repeated clinical examinations. Compared with PBO, MSLT showed that SRC subjects were more vigilant from the onset (P=0.001) to the end of SD (P < 0.0001) whereas some cognitive functions were improved till the thirty third of SD but others were ameliorated through all the SD period and alertness was better from the thirteenth hour of SD, as shown by Stroop's test (P=0.048). We showed that 300-mg SRC given twice daily during a 64-h SD is able to antagonize the impairment produced on vigilance and cognitive functions. PMID- 11903858 TI - The effects of 1 week of REM sleep deprivation on parvalbumin and calbindin immunoreactive neurons in central visual pathways of kittens. AB - Many maturational processes in the brain are at high levels prenatally as well as neonatally before eye-opening, when extrinsic sensory stimulation is limited. During these periods of rapid brain development, a large percentage of time is spent in rapid eye movement (REM) sleep, a state characterized by high levels of endogenously produced brain activity. The abundance of REM sleep in early life and its ensuing decline to lower levels in adulthood strongly suggest that REM sleep constitutes an integral part of the activity-dependent processes that enable normal physiological and structural brain development. We examined the effect of REM sleep deprivation during the critical period for visual development on the development of two calcium-binding proteins that are associated with developmental synaptic plasticity and are found in the lateral geniculate nucleus (LGN) and visual cortex. In this study, REM sleep deprivation was carried out utilizing a computer-controlled, cage-shaking apparatus that successfully suppressed REM sleep. Body weight data suggested that this method of REM sleep deprivation produced less stress than the classical multiple-platform-over-water method. In REM sleep-deprived animals with normal binocular vision, the number of parvalbumin-immunoreactive (PV) neurons in LGN was found to be lower compared with control animals but was not affected in visual cortex. The pattern of calbindin-immunoreactivity (CaB) was unchanged at either site after REM sleep deprivation. Parvalbumin-immunoreactivity develops later than calbindin immunoreactivity in the LGN, and the REM sleep deprivation that we applied from postnatal day 42-49 delayed this essential step in the development of the kitten's visual system. These data suggest that in early postnatal brain development, REM sleep facilitates the usual time course of the expression of PV immunoreactivity in LGN neurons. PMID- 11903859 TI - Mental health, parental rules and sleep in pre-adolescents. AB - A health-sleep model concerning the relationship of mental health and parental rules with time in bed and sleep quality has been developed on the basis of survey data collected from 448 children in the first trimester of the regular school year. The children had a mean age of about 11 years and were attending last two grades of primary school. The relations between mental health characteristics, parental rules concerning sleep, sleep environment, sleep quality and time in bed are analysed using a structural equation model. In addition to a latent variable for 'mental health', two uncorrelated latent variables had to be introduced for sleep to achieve a satisfactory fit. One latent variable is related to sleep quality (restorative sleep), and the other relates to lying awake in bed prior to sleep (awake in bed). Restorative sleep shows a strong relationship with mental health, and awake in bed is related to having an own bedroom. PMID- 11903860 TI - Association between polysomnographic sleep measures and health-related quality of life in obstructive sleep apnea. AB - Many facets of health-related quality of life are diminished in obstructive sleep apnea (OSA) as they are in other chronic medical conditions. We speculated that impairment in health-related quality of life (HRQoL) might result from the fatigue and daytime somnolence associated with the sleep disorder, as an indirect result from the fragmentation of night-time sleep in OSA. Our hypothesis was that sleep fragmentation measures would correlate with poorer HRQoL measured by medical outcomes study (MOS) subscales. Thirty-nine patients with polysomnographically-confirmed OSA participated in this study. Pearson's correlations were performed with the following sleep architecture variables: wake after sleep onset, the total number of brief arousals, the number of respiratory related arousals, the rate of respiratory events per hour, and total sleep time. To our surprise, although the total number of arousals was associated with health distress (r=-0.481, P < 0.005), it did not correlate with any other subscales indicating poorer physical and mental health. The relatively insensitive measure of total sleep time (TST) correlated in the expected direction with most subscales. However, after controlling for age and gender, respiratory disturbance indices (RDI) and/or number of arousals emerged as significantly associated with mobility, cognitive functioning, social functioning, energy and fatigue, and health distress. Our findings suggest that polysomnographic indicators of sleep quality and sleep continuity may be an important influence determining many aspects of HRQoL in OSA patients. PMID- 11903861 TI - Neurological impairments and sleep-wake behaviour among the mentally retarded. AB - The objective of the present study was to evaluate the relationship between the sleep-wake behaviour and neurological impairments among mentally retarded people. The sleep-wake behaviour of 293 mentally retarded subjects living in a rehabilitation center was studied by a standardized observation protocol carried out by trained staff members. The protocol consisted of brief check-ups of the subjects' sleep-wake status at 20-min intervals for five randomly chosen 24-h periods during 4 months. From the raw data five sleep-wake behaviour variables were formed. The data concerning the subject characteristics (age, body mass index (BMI), gender, degree of mental retardation, presence of locomotor disability, that of epilepsy, blindness or deafness and the usage of psychotropic medications) were collected from the medical records. Two main findings emerged: (1) severe locomotor disability, blindness and active epilepsy were found to be independent predictors of increased daytime sleep and increased number of wake sleep transitions and (2) the subjects with a combination of two or all three of these impairments had a significantly more fragmented and abnormally distributed sleep than those with none or milder forms of these impairments. Age, BMI, degree of mental retardation and the studied medications played a minor role in the sleep disturbances of the study population. Finally, deafness was not found to be associated with any of the measured sleep-wake variables. PMID- 11903862 TI - Sleep during Ramadan intermittent fasting. AB - During the month of Ramadan intermittent fasting, Muslims eat exclusively between sunset and sunrise, which may affect nocturnal sleep. The effects of Ramadan on sleep and rectal temperature (Tre) were examined in eight healthy young male subjects who reported at the laboratory on four occasions: (i) baseline 15 days before Ramadan (BL); (ii) on the eleventh day of Ramadan (beginning of Ramadan, BR); (iii) on the twenty-fifth day of Ramadan (end of Ramadan, ER); and (iv) 2 weeks after Ramadan (AR). Although each session was preceded by an adaptation night, data from the first night were discarded. Polysomnography was taken on ambulatory 8-channel Oxford Medilog MR-9000 II recorders. Standard electroencephalogram (EEG), electro-oculogram (EOG) and electromyogram (EMG) recordings were scored visually with the PhiTools ERA. The main finding of the study was that during Ramadan sleep latency is increased and sleep architecture modified. Sleep period time and total sleep time decreased in BR and ER. The proportion of non-rapid eye movement (NREM) sleep increased during Ramadan and its structure changed, with an increase in stage 2 proportion and a decrease in slow wave sleep (SWS) duration. Rapid eye movement (REM) sleep duration and proportion decreased during Ramadan. These changes in sleep parameters were associated with a delay in the occurrence of the acrophase of Tre and an increase in nocturnal Tre during Ramadan. However, the 24-h mean value (mesor) of Tre did not vary. The nocturnal elevation of Tre was related to a 2-3-h delay in the acrophase of the circadian rhythm. The amplitude of the circadian rhythm of Tre was decreased during Ramadan. The effects of Ramadan fasting on nocturnal sleep, with an increase in sleep latency and a decrease in SWS and REM sleep, and changes in Tre, were attributed to the inversion of drinking and meal schedule, rather than to an altered energy intake which was preserved in this study. PMID- 11903863 TI - Sleep and daytime sleepiness in retinitis pigmentosa patients. AB - OBJECTIVE: We examined sleep, daytime sleepiness and the ability to stay awake during the day in patients affected with retinitis pigmentosa (RP), to further delineate the role of photoreceptors in the circadian cycle. METHODS: Twelve individuals diagnosed with RP (40 +/- 8 years) And 12 normally sighted healthy individuals (39 +/- 7 years) matched for age, body mass index (BMI) and sex were selected for the study. Participants had their sleep recorded on two consecutive nights and were monitored on the two following days. On the first day, their ability to stay awake and on the second, their sleep propensity were assessed using the Maintenance of Wakefulness Test (MWT) and the Multiple Sleep Latency Test (MSLT), respectively. Self-report measures were obtained using the Pittsburgh Sleep Quality Index (PSQI), the Epworth Sleepiness Scale (ESS), and the Toronto Hospital Alertness Test (THAT). RESULTS: Subjective daytime sleepiness (ESS: 9 +/- 5 vs. 6 +/- 4, P=0.053) and objectively measured sleep propensity (MSLT: 10 +/- 5 vs. 17 +/- 3 min, P < 0.000) were significantly higher in RP patients than controls, whilst their alertness (THAT: 29 +/- 9 vs. 38 +/- 7, P=0.016) and ability to stay awake (MWT: 21 +/- 9 vs. 29 +/- 2 min, P=0.006) were significantly reduced. Retinitis pigmentosa participants had more disturbed nighttime sleep, with significantly more awakenings (arousal index: 14 +/- 8 vs. 8 +/- 6 h, P=0.039), and tended to have less rapid eye movement (REM) sleep (19 +/- 5 vs. 22 +/- 3%, P=0.094). CONCLUSION: Patients with RP have increased daytime sleepiness, reduced alertness and more disturbed nighttime sleep of poorer quality than their normally sighted counterparts, suggesting an influence of photoreceptor degeneration on the circadian cycle. PMID- 11903864 TI - Clinical and polysomnographic characteristics of 34 patients with Kleine-Levin syndrome. AB - There is only scant information on sleep characteristics and long-term follow-up in patients with Kleine-Levin syndrome (KLS). This study describes the clinical course, results of polysomnography and long-term follow-up in a relatively large group of patients with KLS. During the years 1982-97, we encountered 34 patients (26 males and eight females) with KLS. We were able to obtain the original polysomnographs from 28 males and four females. In 25 patients, data regarding their present state of health were obtained. Fourteen agreed to be present at a detailed interview and examination while 11 gave the information by phone. The mean age at onset was 15.8 +/- 2.8 years and the mean diagnostic delay, 3.8 +/- 4.2 years. The mean duration of a single hypersomnolent attack was 11.5 +/- 6.6 days. The main abnormal findings extracted out of 35 polysomnographs obtained from 32 patients during and/or in-between attacks included: decreased sleep efficiency, and frequent awakenings from sleep stage 2. All 25 patients reported present perfect health, with no evidence of behavioral or endocrine dysfunction. In adolescents with periodic hypersomnia, the diagnosis of KLS should be explored. Sleep recordings during a hypersomnolent period will often show frequent awakenings from sleep stage 2. The long-term prognosis is excellent. PMID- 11903865 TI - Veterinary pharmacovigilance: between regulation and science. AB - Veterinary pharmacovigilance has shown a remarkable development in recent years. After presenting briefly the regulatory context, this paper considers the scope of veterinary pharmacovigilance, causality assessment of suspected adverse drug reactions (ADRs), the systems existing in different countries and the evolution for the coming years. The involvement of veterinary research and teaching institutions should certainly become more important in this cross-road activity. PMID- 11903866 TI - Pharmacokinetic behavior of marbofloxacin after intravenous and intramuscular administrations in adult goats. AB - The pharmacokinetic behavior of marbofloxacin was studied in goats after single dose intravenous (i.v.) and intramuscular (i.m.) administrations of 2 mg/kg bodyweight. Drug concentration in plasma was determined by high performance liquid chromatography (HPLC) and the data collected were subjected to compartmental and noncompartmental kinetic analysis. This compound presented a relatively high volume of distribution (Vss=1.31 L/kg), which suggests good tissue penetration, and a total body clearance (Cl) of 0.23 L/kg small middle doth, which is related to a long elimination half-life (t1/2beta=7.18 h and 6.70 h i.v. and i.m., respectively). Pharmacokinetic parameters were not significantly different between both routes of administration. Marbofloxacin was rapidly absorbed after i.m. administration (Tmax=0.9 h) and had high bioavailability (F=100.74%). PMID- 11903867 TI - A pharmacokinetic study of amoxycillin in febrile beagle dogs following repeated administrations of endotoxin. AB - The pharmacokinetics of amoxycillin was studied in nine male beagle dogs under healthy and febrile conditions. In Period 1, dogs received 20 mg/kg of an oral suspension of amoxycillin. Intravenous doses of saline, 2 and 20 microg/kg of endotoxin (LPS from Escherichia coli serotype) were administered to dogs (three per group) prior to administration of 20 mg/kg of amoxycillin in Period 2. Rectal temperature and behavioral changes were recorded and blood samples were collected over 12 h for pharmacokinetic analysis. Amoxycillin was assessed in plasma using liquid chromatography coupled with mass spectrometry. Plasma concentrations were analysed using a one-compartment model with lag-time for absorption using an iterative two-stage method. As compared with control groups, amoxycillin clearance decreased significantly with preliminary treatments of 2 microg/kg endotoxin (0.209 vs. 0.140 L/h kg, P < 0.05) and 20 microg/kg endotoxin (0.214 vs. 0.075 L/h kg, P < 0.05). As a result of this, the area under curve for the 2 and 20 microg/kg endotoxin groups increased significantly 100.4 vs. 149.4 microg h/mL (P < 0.05) and 99.2 vs. 277.7 microg h/mL (P < 0.05), respectively. Other drugs currently used for the treatment of fever and septic shock should be re evaluated using a febrile animal model to avoid improper dose administration. PMID- 11903869 TI - Morphine, pethidine and buprenorphine disposition in the cat. AB - Pharmacokinetics of morphine, buprenorphine and pethidine were determined in 10 cats. Six cats received morphine (0.2 mg/kg) intravenously and four intramuscularly. Five received buprenorphine (0.01 mg/kg) intravenously and six intramuscularly. Six received pethidine (5 mg/kg) intramuscularly. Jugular venous blood samples were collected at time points to 24 h, and plasma morphine concentrations were measured by high performance liquid chromatograpy (HPLC), buprenorphine by radioimmunoassay (RIA) and pethidine by gas chromatography. Our data for morphine show elimination half-life (t1/2el) 76.3 min intravenous (i.v.) and 93.6 min intramuscular (i.m.); mean residence time (MRT) 105.0 and 120.5 min; clearance (Clp) 24.1 and 13.9 mL/kg/min; and volume of distribution (V(dss)) 2.6 and 1.7 L/kg, respectively. Comparable data for buprenorphine are t1/2el 416.8 and 380.2 min; MRT 417.6 and 409.8 min; Clp 16.7 and 23.7 mL/kg/min; and V(dss) 7.1 and 8.9 L/kg. For i.m. pethidine, t1/2el 216.4 min; MRT 307.5 min; Clp 20.8 mL/kg/min and V(dss) 5.2 L/kg. For i.m. dosing, the tmax for morphine, buprenorphine and pethidine were 15, 3 and 10 min, respectively. The pharmacokinetics of the three opioids in cats are broadly comparable with those of the dog, although there is a suggestion that the cat may clear morphine more slowly. PMID- 11903868 TI - An assessment of antimicrobial consumption in food producing animals in Kenya. AB - Antimicrobial agents are useful for control of bacterial infections in food animals and man. Their prudent use in these animals is important to control any possible development and transfer of resistance between animals and man. The objective of this study was to generate quantitative information to evaluate antimicrobial usage patterns by animal species, route of administration, antimicrobial class and type of use from 1995 to 1999 in Kenya. Theses data are essential for risk analysis and planning and can be helpful in interpreting resistance surveillance data, and evaluating the effectiveness of prudent use efforts and antimicrobial resistance mitigation strategies. Data on quantities of active substance classes were collected from the official records of the Pharmacy and Poisons Board of the Ministry of Health and analysed in MS Excel 2000 program. The mean antimicrobial consumption for the 5-year period was 14 594 +/- 1457 kg per year. This was distributed in the various antimicrobial classes as follows: 7975 kg (54.65%) of tetracyclines, 3103.96 kg (21.27%) of sulfonamides and 954.5 kg (6.56%) of aminoglycosides, 905 kg (6.20%) of beta-lactams, 94 kg (0.64%) of quinolones, 35 kg (0.24%) of macrolides and 24 kg (0.16%) of others (tiamulin). Mean consumption per year among the various food animals was: 10 989 +/- 357 kg in large animals (cattle, sheep, pigs and goats), 2906 +/- 127 kg in poultry alone and 699 +/- 427 kg in both large animals and poultry. These quantities represented 56.56% (8255 kg) consumption per year for parenteral use, 41.79% (6098 kg) for oral use and 1.65% (241 kg) for topical use (intramammary and eye ointments) in cattle. With respect to intended use in food producing animals, the mean consumption per year was: 13 178 kg (90.30%) for therapeutic use (ST), 4 kg (0.03%) for prophylactic treatment (PT) and 1411 +/- 246 kg (9.67%) was used both for therapeutic and prophylactic purposes (GPT). The study confirmed that antimicrobials are not used for growth promotion in Kenya. There was no specific trend in the quantities of active antimicrobial classes. This study has revealed that the tetracyclines, sulfonamides and trimethoprim, nitrofurans aminoglycosides, beta-lactams and the quinolones are the most commonly used drugs in food-producing animals in Kenya. Tetracyclines contributed approximately 55% of the total consumption, and there was an increasing trend in the consumption of quinolones from 1998. PMID- 11903870 TI - Oxidative monensin metabolism and cytochrome P450 3A content and functions in liver microsomes from horses, pigs, broiler chicks, cattle and rats. AB - The oxidative metabolism of monensin, an ionophore antibiotic extensively used in veterinary practice as a coccidiostat and a growth promoter, was studied in hepatic microsomal preparations from horses, pigs, broiler chicks, cattle and rats. As assayed by the measurement of the amount of the released formaldehyde, the rate of monensin O-demethylation was nearly of the same order of magnitude in all species, but total monensin metabolism, which was estimated by measuring the rate of substrate disappearance by a high-performance liquid chromatography (HPLC) method, was highest in cattle, intermediate in rats, chicks and pigs, and lowest in horses. When expressed as turnover number (nmol of metabolized monensin/min nmol cytochrome P450-1), the catalytic efficiency (chick >> cattle >> pig approximately rat > horse) was found to correlate inversely with the well known interspecies differences in the susceptibility to the toxic effects of the ionophore, which is characterized by an oral LD50 of 2-3 mg/kg bodyweight (bw) in horses, 50-80 mg/kg bw in cattle and 200 mg/kg bw in chicks. Chick and cattle microsomes also displayed both the highest catalytic efficiency toward two P450 3A dependent substrates (erythromycin and triacetyloleandomycin) and the highest immunodetectable levels of proteins cross-reacting with anti rat P450 3A1/2. Further studies are required to define the role played by this isoenzyme in the oxidative biotransformation of the drug in food producing species. PMID- 11903871 TI - Continuous measurement of caffeine and two metabolites in blood and skeletal muscle of unrestrained adult horses by semi-automated in vivo microdialysis. AB - Concentrations of caffeine (CA) and two metabolites were measured simultaneously in venous blood and splenius muscle of adult horses using a semi-automated in vivo microdialysis sampling technique. Dialysates from muscle and jugular vein were collected continuously for 48 h and drug levels were determined by high performance liquid chromatography (HPLC). Following i.v. injection, CA (3 mg/kg) attained a peak blood level of nearly 5400 +/- 600 ng/mL and decreased with a half-life of 15.3 +/- 0.7 h. Pharmacokinetic and statistical comparisons between CA concentrations in jugular dialysates and plasma samples revealed no significant differences between these sampling techniques. However, measurements in muscle and blood revealed unexpected pharmacokinetic differences, including significantly elevated concentrations of CA in muscle for 4 h following drug administration. In contrast, the CA metabolites theophylline (TP) and theobromine (TB) exhibited delayed appearances in muscle and blood with peak concentrations of 300 +/- 60 ng/mL (TP) and 150 +/- 50 ng/mL (TB) detected in both tissues 1 day following CA administration. This study demonstrates that our novel semi automated microdialysis procedure for continuous monitoring of drug and metabolite levels may be useful for related studies in other domesticated large animal species. PMID- 11903872 TI - Acetaminophen UDP-glucuronosyltransferase in ferrets: species and gender differences, and sequence analysis of ferret UGT1A6. AB - The principal objective of this study was to determine whether ferrets glucuronidate acetaminophen more slowly compared with other species, and if so investigate the molecular basis for the difference. Acetaminophen-UDP glucuronosyltransferase (UGT) activities were measured using hepatic microsomes from eight ferrets, four humans, four cats, four dogs, rat, mouse, cow, horse, monkey, pig and rabbit. Gender differences between male and female ferret livers were explored using enzyme kinetic analysis. Immunoblotting of microsomal proteins was also performed using UGT-specific antibodies. Finally, the exon 1 region of UGT1A6, a major acetaminophen-UGT, was sequenced. Glucuronidation of acetaminophen was relatively slow in ferret livers compared with livers from all other species except cat. Gender differences were also apparent, with intrinsic clearance (Vmax/Km) values significantly higher in male compared with female ferret livers. Furthermore, Vmax values correlated with densitometric measurements of two protein bands identified with a UGT1A subfamily-specific antibody. No deleterious mutations were identified in the exon 1 or flanking regions of the ferret UGT1A6 gene. In conclusion, like cats, ferret livers glucuronidate acetaminophen relatively slowly. However, unlike cats, in which UGT1A6 is encoded by a pseudogene and dysfunctional, there are no defects in the ferret UGT1A6 gene which could account for the low activity. PMID- 11903873 TI - Effects of altered plasma alpha-1-acid glycoprotein levels on pharmacokinetics of some basic antibiotics in pigs: simulation analysis. AB - Effects of altered plasma alpha-1-acid glycoprotein (AGP) levels on pharmacokinetic parameters of basic antimicrobials, erythromycin (EM), lincomycin (LM) and clindamycin (CM) were evaluated in pigs by simulation analysis. Intravenous (i.v.) injections of EM, LM and CM were performed to obtain pharmacokinetic parameters in healthy conditions. Binding parameters were obtained from an in vitro study using ultrafiltration. Simulation studies indicated that an increase of plasma AGP levels resulted in a decrease of both volume of distribution at steady state (Vdss) and total body clearance (Cltot) for all the drugs. Elimination rate constant for LM was almost unchanged by an increase of plasma AGP levels, whereas those for EM and CM were increased. Plasma concentration-time profiles at a high AGP level (often observed in pathophysiological conditions) were also simulated. All of the total plasma concentration-time profiles were different from those at normal AGP level. The differences were characterized by a higher initial concentration with faster or similar elimination. Unbound plasma concentration-time profile of LM was unaffected by AGP levels, whereas EM and CM were eliminated from plasma more rapidly at high AGP level. These results suggested that adjustment of dosage regimen of EM and CM is required in pathophysiological conditions, but that of LM is not required. PMID- 11903874 TI - Comparative studies on Polyferm and Fermosorb, two oral (ferment + sorbent) - type preparations designed for therapy/prophylaxis of intestinal infections in animal neonates. AB - Polyferm and Fermosorb are oral acid resistant antimicrobial enzyme preparations designed specifically for therapy/prophylaxis of intestinal infections in animal neonates. Both are authorized for use throughout the former Soviet Union, but until now only Fermosorb is being applied on a large scale. The comparative studies on these two preparations, described in this paper, were carried out in order to find differences between the preparations. Characteristics that were compared included stability of the preparations in acidic environment as well as in storage (in vitro studies), and their efficacy for the treatment and prophylaxis of colibacillosis in newborn calves (in vivo studies). Results of in vitro studies revealed that proteolytic enzymes of Polyferm (as well as lytic enzymes of Fermosorb) were suitably (and in a very similar magnitude) protected from the influence of the acidic environment. The complete enzyme activity retention period in storage at room temperature of Polyferm and Fermosorb was equally high (5 years). In vivo studies performed on 2000 calves revealed that both preparations were highly effective and, although the efficacy of Polyferm was a bit lower than that of Fermosorb (93.6% vs. 95.0%, 94.6% vs. 95.8% for therapy and prophylaxis of colibacillosis, respectively), no statistically significant differences in the number of Polyferm vs. Fermosorb cured/protected animals were found. It is concluded that there were no reasons, other than the lack of supportive advertising materials, that might impede the utility of Polyferm. PMID- 11903875 TI - Oral bioavailability of levamisole in goats. PMID- 11903876 TI - Chiral pharmacokinetics of ketorolac in sheep after intravenous and intramuscular administration of the racemate. PMID- 11903877 TI - Inadequacy of low-volume resuscitation with hemoglobin-based oxygen carrier hemoglobin glutamer-200 (bovine) in canine hypovolemia. PMID- 11903879 TI - Update on hepatic stem cells. AB - The liver, like most organs in an adult healthy body, maintains a perfect balance between cell gain and cell loss. Though normally proliferatively quiescent, simple hepatocyte loss such as that caused by partial hepatectomy, uncomplicated by virus infection or inflammation, invokes a rapid regenerative response to restore liver mass. This restoration of moderate cell loss and 'wear and tear' renewal is largely achieved by hepatocyte self-replication. Furthermore, cell transplant models have shown that hepatocytes can undergo significant clonal expansion. Such observations indicate that hepatocytes are the functional stem cells of the liver. More severe liver injury activates a facultative stem cell compartment located within the intrahepatic biliary tree, giving rise to cords of biliary epithelia within the lobules before these cells differentiate into hepatocytes. A third population of stem cells with hepatic potential resides in the bone marrow; these haematopoietic stem cells can contribute to the albeit low renewal rate of hepatocytes, make a more significant contribution to regeneration, and even completely restore normal function in a murine model of hereditary tyrosinaemia. How these three stem cell populations integrate to achieve a homeostatic balance is not understood. This review focuses on three aspects of liver stem cell biology: 1) the hepatic stem cell candidates; 2) models of cell transplantation into the liver; and 3) the therapeutic potential of hepatic stem cells. PMID- 11903880 TI - Neuronal nitric oxide synthase immunoreactivity in the guinea-pig liver: distribution and colocalization with neuropeptide Y and calcitonin gene-related peptide. AB - AIMS/BACKGROUND: The innervation pattern of the guinea-pig liver is similar to that of the human liver. However, many aspects of the distribution of the neuronal isoform of the enzyme nitric oxide synthase (nNOS) in the guinea-pig liver and its colocalization with neuropeptides remain to be elucidated. METHODS: The distribution of nNOS was studied in fixed guinea-pig liver by light microscopic immunohistochemistry. Confocal analysis was used to determine its colocalization with neuropeptide Y (NPY) or calcitonin gene-related peptide (CGRP). RESULTS: nNOS-immunoreactive (nNOS-IR) nerves were observed in relation to hilar and interlobar vessels and in Glisson's capsule. A few nNOS-IR ganglia were observed in the extrahepatic bile duct and close to the interlobar portal triads. In addition, nNOS-IR fibers were located in the interlobular portal triads and pervading the parenchyma. Moreover, nNOS-IR nerves were demonstrated for the first time in the larger central veins and in the hepatic vein. nNOS-NPY and nNOS-CGRP colocalizations were detected in the fibromuscular layer of the bile duct and periductal plexus, respectively. CONCLUSIONS: These results support the phylogenetic conservation of the nNOS-IR hepatic innervation and its possible contribution to the regulation of hepatic blood flow and certain hepatic functions. PMID- 11903881 TI - Presence of low levels of anti-HBs antibody in so-called 'anti-HBc alone' subjects. AB - BACKGROUND/AIMS: The 'anti-Hbc alone' pattern could sometimes be that of subjects who produced anti-HBs after recovery, but at a lower level than that detectable using commercial assays. This study aimed to test this hypothesis. METHODS: A total of 104 'anti-HBc alone' serum samples, i.e.positive for the anti-HBc antibody but not for HBsAg nor for anti-HBs antibody, were recruited when routine testing a broad population of employees, patients and pregnant women from a university hospital. A possible subliminal anti-HBs production, that would have escaped detection by commercial EIAs, was investigated by comparing the optical densities (ODs) obtained in vaccinees (commercial anti-HBs EIA) to those of a control group of 100 nonimmunised and nonvaccinated subjects. RESULTS: The median OD was significantly higher (p<0.0001) in the 'anti-HBc alone' subjects (OD=0.035) than in the controls (OD=0.023). Thirty-six percent of the 'anti-HBc alone' subjects had an anti-HBs OD higher than the median OD of the controls+2SD. 'Anti-HBc alone' subjects with anti-HBe antibody had higher anti-HBs ODs (0.041) than had those without anti-HBe (0.029). In 'anti-HBc alone' subjects, the anti HBs ODs, although under the cut-off value of the EIA, were found to be higher than in the controls. CONCLUSION: Our results show low anti-HBs production in some of the subjects studied. PMID- 11903882 TI - Effects of salviainolic acid A (SA-A) on liver injury: SA-A action on hepatic peroxidation. AB - BACKGROUND/AIMS: This study aimed to investigate the actions of salviainolic acid A (SA-A), an antiperoxidative component of Salvia miltiorrhiza (Sm), on rat liver injury and fibrosis. METHODS: Acute and chronic rat liver injury models were established using carbon tetrachloride (CCl4). After 48 h (acute) or during 6 weeks of CCl4 injection, rats were further divided and treated with biphenyl dimethyl-dicarboxylate (BDD) or colchicine, as a control antifibrotic treatment, with Sm, a herbal compound, or SA-A, a water-soluble extract of Sm. Liver function was investigated by assessing alanine transaminase (ALT) and aspartate transaminase (AST) activities, histological analysis, hydroxyproline (Hyp) and malondiadehyde (MDA) content. In vitro, isolated cultured hepatocytes were injured with CCl4 gas for 24 h, followed by treatment with either vitamin E or various concentrations of SA-A. The extent of hepatocyte injury was monitored by analyzing various lipid peroxidative parameters such as superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), lactase dehydrogenase (LDH), and glutathione peroxidase (GSH-PX) levels in hepatocyte supernatants. RESULTS: SA-A significantly decreased abnormal serum ALT activity both in acutely and chronically injured rat livers, decreased abnormal serum AST activity, Hyp and MDA content and attenuated hepatic collagen deposition. After CCl4 incubation and injury, the activities of AST, ALT CAT, GSH-PX and LDH and MDA content in hepatocyte supernatants increased significantly, but GSH levels decreased significantly. SA-A markedly improved these pathological changes in a dose dependent manner. 10(-4) mol/l SA-A had stronger inhibitory action than vitamin E. CONCLUSIONS: Our studies suggest that SA-A has antiperoxidative effects on injured hepatocytes in liver injury and fibrosis induced by CCl4. PMID- 11903883 TI - Large-needle biopsy versus thin-needle biopsy in diagnostic pathology of liver diseases. AB - BACKGROUND/AIMS: A study was carried out to determine whether thin-needle biopsy (TNB) yields enough material to study diffuse liver diseases. METHODS: Using TNB (20G and 21G) and a conventional Menghini-type large-needle biopsy (LNB; 17G), the amounts of tissue obtained and the histopathological diagnoses were compared. The biopsies were obtained by surgeons with a several-stroke method (17GS) and by physicians with a single-strike method (17GP, 20GP, 21GP). A total of 343 biopsy specimens from 258 patients were included in the study. RESULTS: A comparison of the mean values for the length of the core biopsy, as well as the mean numbers of portal tracts and terminal hepatic veins among the four groups showed significant differences (p<0.001): the mean number of portal tracts obtained with 17GS was 13.8+/-6.5, with 17GP it was 9.7+/-5.9, with 20GP it was 6.7+/-4.4, and with 21GP it was 4.0+/-3.1. A comparison of the histopathological diagnoses showed no major differences between 17GP and 20GP; the diversity and frequencies of the diagnoses were similar. CONCLUSION: We suggest that the use of TNB, particularly 20G-size needles, could be extended to the investigation of diffuse liver diseases in which LNB carries a high risk of complications or is contraindicated, and when the diagnosis is the primary reason for the biopsy rather than grading or staging of a known diffuse disease. PMID- 11903884 TI - Meta-analysis of propylthiouracil for alcoholic liver disease--a Cochrane Hepato Biliary Group Review. AB - AIMS/BACKGROUND: The aim of this review was to determine the benefits and adverse effects of propylthiouracil for patients with alcoholic liver disease. METHODS: Systematic Cochrane Review of randomised clinical trials. The Cochrane Hepato Biliary Controlled Clinical Trials Register, The Cochrane Library, MEDLINE, and full text searches were combined. All analyses were performed according to the intention-to-treat method. Only randomised clinical trials studying patients with alcoholic steatosis, alcoholic fibrosis, alcoholic hepatitis and/or alcoholic cirrhosis were included. Interventions encompassed propylthiouracil at any dose versus placebo or no intervention. The trials could be double-blind, single-blind or unblinded. RESULTS: Six randomised clinical trials randomising 710 patients demonstrated no significant effects of propylthiouracil versus placebo on mortality (Peto odds ratio (OR) 0.91, 95% confidence interval (CI) 0.59 to 1.40), liver-related mortality (OR 0.78, CI 0.45 to 1.33), complications to the liver disease (OR 1.14, CI 0.58 to 2.24), and liver histology. Propylthiouracil was associated with a nonsignificant trend toward an increased risk of nonserious adverse events (OR 1.49, CI 0.74 to 2.99) and with the rare occurrence of serious adverse events (leukopenia). CONCLUSIONS: This systematic review could not demonstrate any significant effect of propylthiouracil on any clinically important outcomes (mortality, liver-related mortality, liver complications and liver histology) of patients with alcoholic liver disease. PMID- 11903885 TI - Insulin-like growth factor-I restores the reduced somatostatinergic tone controlling growth hormone secretion in cirrhotic rats. AB - BACKGROUND/AIMS: An altered growth hormone/insulin-like growth factor-I (GH/IGF I) axis occurs in advanced liver cirrhosis, characterised by diminished serum levels of IGF-I and increased concentrations of GH. Under normal conditions, GH release is mediated by somatostatin (SS) inhibition. However, the influence of SS on GH release in cirrhosis is not well known. IGF-I supplementation has beneficial effects in experimental cirrhosis, and - under physiological conditions - IGF-I increases SS, inhibiting GH. The aims of this work were to study SS tone in cirrhotic animals and to evaluate whether IGF-I treatment influences SS tone, controlling GH secretion in cirrhosis. METHODS: We studied the influence of SS on GH secretion by assessing GH response to pyridostigmine (PD) in cirrhotic rats treated and untreated with IGF-I. Liver cirrhosis was induced with CCl4-inhalation for 11 weeks in male Wistar rats. The animals were randomly divided into two groups: CI+IGF (n=12), which received IGF-I treatment for 12 days (2 microg/100 g body wt-1 x d-1) and CI (n=12), which received saline. Healthy controls (CO, n=12) were studied at the same time. On day 13, animals from each group were subdivided into two groups (n=6) in order to explore the effect of a PD intrajugular bolus (10 microg x 100 gbw-1) on serum GH levels (at 0,10,20,30 and 60 min), which were assessed by RIA. RESULTS: PD bolus did not exert any effect on GH serum levels in the CI group, suggesting a low SS tone in cirrhotic rats. However, PD induced an increase in GH levels into CO and CI+IGF groups. In conclusion, as occurs under normal conditions, the cholinergic system is a significant modulator of GH secretion in experimental liver cirrhosis. CONCLUSION: Cirrhotic rats have a reduced somatostatinergic tone which can be restored by IGF-I supplementation, suggesting that somatostatin is the main factor involved in the feed-back regulation between GH and IGF-I in cirrhosis. PMID- 11903886 TI - Minimal liver injury in chronic hepatitis C virus infection is associated with low levels of soluble TNF-alpha/Fas receptors and acquisition in childhood. AB - BACKGROUND/AIMS: The rate of progression to cirrhosis of chronic hepatitis C might be related to an upregulation of TNF-alpha/Fas pathways. METHODS: The serum levels of soluble TNF-alpha type II receptor (sTNFr-II) and soluble Fas antigen (sFas) were analyzed in patients with different histological outcomes of chronic parenterally acquired HCV infection of similar duration. RESULTS: One hundred and forty-five HCV-infected patients had a known duration of infection. Twelve (8.3%) patients had minimal changes and were assigned to the case group. The control group was selected from the 24 (17%) patients with cirrhosis and the 54 (37%) with chronic active hepatitis (CAH). Two controls, one with CAH and one with cirrhosis, were paired with the cases following these criteria: duration of infection, transmission route and sex. The proportions of genotype 1b and HCV RNA serum levels were similar among the groups. The median serum levels of sTNFr-II and sFas were significantly lower in the case group than in the control groups. The cases were significantly younger when they became infected than the control groups. Indeed, most cases were infected within the first 10 years of life. sTNFr II and age at infection were independently associated with the minimal injury case group. When sTNFr-II was excluded from the logistic regression model, sFas and age at infection were independently associated with the case group. CONCLUSION: The rate of progression of parenterally acquired chronic hepatitis C to end-stage liver disease might be related to an upregulation of the TNF alpha/Fas pathways and an age-dependent host response. PMID- 11903887 TI - Sinusoidal endothelial cell injury by superoxide anion and iron in the Propionibacterium acnes-pretreated and lipopolysaccharide-stimulated rat liver. AB - AIMS/BACKGROUND: We attempted to measure the generation of superoxide anion, examine its site of release and determine its pathological role in Propionibacterium acnes-lipopolysaccharide-induced liver injury in the rat. METHODS: The P. acnes-pretreated (16 mg/kg i.v.) rat liver was perfused with buffer containing lipopolysaccharide (2.5 microg/ml). Chemiluminescence enhanced with Cypridina luciferin analog, MCLA, and reduction of nitro blue tetrazolium were used for detecting superoxide anion. Leakage of enzymes and release of cytokines into the perfusate, and histological specimens were also examined. RESULTS: Superoxide dismutase-inhibitable chemiluminescence peaked at 30 min of lipopolysaccharide infusion and blue formazan precipitate was histochemically deposited mainly on hepatic macrophages. Purine nucleoside phosphorylase (PNP) activity in the perfusate, as a marker of sinusoidal endothelial cell injury, reached its maximum at 50 min and aspartate aminotransferase (AST) activity, as a marker of hepatocyte injury, reached a plateau at 90 min. Simultaneous treatment with superoxide dismutase and deferoxamine mesylate significantly suppressed the leakage of PNP and AST. Release of tumor necrosis factor-alpha and growth-related oncogene/cytokine-induced neutrophil chemoattractant-1 lagged behind PNP leakage. Light microscopy showed destruction of the sinusoids followed by hepatocyte necrosis. Electron microscopy revealed adherence of hepatic macrophages to sinusoidal endothelial cells. CONCLUSION: These results indicate that superoxide anion released from hepatic macrophages may induce sinusoidal endothelial cell injury via interaction with iron in the P. acnes-lipopolysaccharide-treated liver. PMID- 11903888 TI - Adaptive evolutionary conservation: towards a unified concept for defining conservation units. AB - Recent years have seen a debate over various methods that could objectively prioritize conservation value below the species level. Most prominent among these has been the evolutionarily significant unit (ESU). We reviewed ESU concepts with the aim of proposing a more unified concept that would reconcile opposing views. Like species concepts, conflicting ESU concepts are all essentially aiming to define the same thing: segments of species whose divergence can be measured or evaluated by putting differential emphasis on the role of evolutionary forces at varied temporal scales. Thus, differences between ESU concepts lie more in the criteria used to define the ESUs themselves rather than in their fundamental essence. We provide a context-based framework for delineating ESUs which circumvents much of this situation. Rather than embroil in a befuddled debate over an optimal criterion, the key to a solution is accepting that differing criteria will work more dynamically than others and can be used alone or in combination depending on the situation. These assertions constitute the impetus behind adaptive evolutionary conservation. PMID- 11903889 TI - How mitochondrial DNA diversity can help to understand the dynamics of wild cultivated complexes. The case of Medicago sativa in Spain. AB - In order to clarify the relationships (genetic exchange and shared ancestry) between natural and cultivated populations of alfalfa (Medicago sativa L.) in Spain, we investigated the patterns of mitochondrial DNA variation (characterized through restriction fragment length polymorphism) for 248 individuals in seven natural and six cultivated populations of this species. Mitochondrial variation was evidenced in both natural and cultivated populations of M. sativa. Among the seven mitotypes identified in the species, two were specific of the natural populations, a result attesting the fact that the Spanish wild form of M. sativa is an original genetic pool compared to the cultivated one. Other mitotypes were observed in both natural and cultivated populations, suggesting the occurrence of gene flow through seeds from cultivated towards natural populations. Comparisons with previously gathered nuclear and phenotypic data give insights into the different evolutionary forces acting on the different kinds of Spanish natural populations examined so far. PMID- 11903890 TI - MHC class II beta sequence diversity in the deer mouse (Peromyscus maniculatus): implications for models of balancing selection. AB - We studied population polymorphism at a major histocompatibility complex (MHC) class II beta gene in the deer mouse (Peromyscus maniculatus). We found that: (i) a single population of P. maniculatus has significantly higher levels of DNA and protein sequence diversity than worldwide samples from homologous genes in other taxa, including humans and mice; and (ii) the genealogy of allelic sequences in P. maniculatus deviates significantly from theoretical expectation under a model of symmetric balancing selection, in that alleles are relatively more divergent than expected. We suggest that the observation of high levels of pairwise allelic sequence divergence and deviation of the genealogy from theoretical expectation in P. maniculatus together provide support for a divergent allele advantage model for the maintenance of MHC polymorphism. PMID- 11903891 TI - Glacial biogeography of North American coho salmon (Oncorhynchus kisutch). AB - To study the glacial biogeography of coho we examined 20 microsatellite loci and mitochondrial DNA D-loop sequence in samples from Alaska to California. Microsatellite data divided our samples among five biogeographic regions: (1) Alaska and northern coastal British Columbia; (2) the Queen Charlotte Islands; (3) the mainland coast of British Columbia and northern Washington State; (4) the Thompson River; and (5) Oregon and California. The D-loop sequence data suggested three geographical regions: (1) Oregon and California; (2) the Thompson River; and (3) all the other sites north of the southern ice margin. Microsatellite data revealed no difference in the number of alleles in different regions, but mitochondrial DNA data revealed a cline of decreasing diversity from south to north. We suggest that the two signals presented by these different marker types illuminate two time frames in the history of this species. Endemic microsatellite diversity in Alaska and on the Queen Charlotte Islands provides evidence in favour of Fraser Glaciation refugia in these regions. The loss of mitochondrial variation from south to north suggests that one of the earlier, more extensive, Pleistocene glaciations eliminated coho from its northern range. PMID- 11903892 TI - Prey specialization may influence patterns of gene flow in wolves of the Canadian Northwest. AB - This study characterizes population genetic structure among grey wolves (Canis lupus) in northwestern Canada, and discusses potential physical and biological determinants of this structure. Four hundred and ninety-one grey wolves, from nine regions in the Yukon, Northwest Territories and British Columbia, were genotyped using nine microsatellite loci. Results indicate that wolf gene flow is reduced significantly across the Mackenzie River, most likely due to the north south migration patterns of the barren-ground caribou herds that flank it. Furthermore, although Banks and Victoria Island wolves are genetically similar, they are distinct from mainland wolf populations across the Amundsen Gulf. However, low-level island-mainland wolf migration may occur in conjunction with the movements of the Dolphin-Union caribou herd. Whereas previous authors have examined isolation-by-distance in wolves, this study is the first to demonstrate correlations between genetic structure of wolf populations and the presence of topographical barriers between them. Perhaps most interesting is the possibility that these barriers reflect prey specialization by wolves in different regions. PMID- 11903893 TI - Nucleotide diversity in populations of balitorid cave fishes from Thailand. AB - Genetic variabilities in four cave and eight surface species of balitorid freshwater fishes from Thailand were assessed using random amplified polymorphic DNA (RAPD). Cave species have consistently lower RAPD variation than surface species and it is hypothesized that this difference is a function of reduced population size in cave fishes. Indices of nucleotide diversity (pi) were calculated from the RAPD data and are four to five times higher for the surface species than the cave species: 6.4 x 10-3 vs. 1.4 x 10-3. The pi-values for cave fishes are significantly lower than those for surface balitorids and those measured in other species using RAPD/amplified fragment length polymorphism data. PMID- 11903894 TI - Social and genetic characteristics of geographically isolated populations in the ant Formica cinerea. AB - The ant Formica cinerea in northern Europe has geographically isolated populations that were examined using five microsatellite loci. The populations differ widely regarding the social organization of colonies. Based on genetic relatedness (r) among worker nest mates, the populations were classified as M type with monogynous (single queen) colonies (r > 0.59), as P type with polygynous colonial networks (r < 0.1), or as intermediate with weakly polygynous colonies (0.16 < r < 0.47). The social types showed weak geographical clustering, but the overall distribution indicated that the shift between the social types has occurred several times. The geographically isolated populations had slightly reduced levels of genetic diversity compared to populations from areas where the species is abundant and continuously distributed. Many of the isolated populations consisted of monogynous or weakly polygynous colonies, making their effective population sizes small, and some of them also showed weak bottleneck effects. The overall level of microsatellite diversity within populations was relatively high and differentiation among populations low, indicating recent connections. Isolation of populations may thus be a new phenomenon resulting from reduction of suitable habitats. At the local level, we obtained limited support from a group of nearby subpopulations in southern Finland to the hypothesis that the P type is connected to restricted dispersal. Other P type populations did not, however, show similar elevated levels of differentiation. PMID- 11903895 TI - A hybrid zone between hydrothermal vent mussels (Bivalvia: Mytilidae) from the Mid-Atlantic Ridge. AB - This study provides the first example of a hybrid zone between animal taxa distributed along the mid-ocean ridge system. We examined the distribution and genetic structure of deep-sea hydrothermal vent mussels (Bivalvia: Mytilidae) along a 2888-km portion of the Mid-Atlantic Ridge between 37 degrees 50' N and 14 degrees 45' N latitude. Mitochondrial DNA (mtDNA), allozymes and multivariate morphometric evidence discriminated between individuals of a northern species, Bathymodiolus azoricus, and a southern species, B. puteoserpentis, that were separated by an intermediate ridge segment almost devoid of mussels. A small sample of mussels from Broken Spur, a vent locality along this intermediate zone, revealed a mixed population with gene frequencies and morphology that were broadly intermediate to those of the northern and southern species. Multilocus clines in mtDNA and allozyme frequencies were centred over the intermediate zone. We consider intrinsic and extrinsic processes that might limit genetic exchange across this hybrid zone. PMID- 11903896 TI - Extensive trans-species mitochondrial polymorphisms in the carabid beetles Carabus subgenus Ohomopterus caused by repeated introgressive hybridization. AB - To study the potential importance of introgressive hybridization to the evolutionary diversification of a carabid beetle lineage, we studied intraspecific and trans-species polymorphisms in the mitochondrial NADH dehydrogenase subunit 5 (ND5) gene sequence (1083 bp) in four species of the subgenus Ohomopterus (genus Carabus) in central and eastern Honshu, Japan. Of the four species, C. insulicola is parapatric with the other three, and can hybridize naturally with at least two. This species possesses two haplotypes of remote lineages. We classified ND5 haplotypes using polymerase chain reaction restriction fragment length polymorphism with TaqI endonuclease for 524 specimens, and sequenced 143 samples. Analysis revealed that each species was polyphyletic in its mitochondrial DNA phylogeny, representing a marked case of trans-species polymorphism. Recent one-way introgression of mitochondria from C. arrowianus nakamurai to C. insulicola, and from C. insulicola to C. esakii, was inferred from the frequency of identical sequences between these species and from direct evidence of hybridization in their contact zones. Other intraspecific polymorphisms in the four species may be due to undetected introgressive hybridization (e.g. C. insulicola to C. maiyasanus) or from stochastic lineage sorting of ancestral polymorphisms. This beetle group has a genital lock-and-key system, with species-specific or subspecies-specific genital morphology that may act as a barrier to hybridization. However, our results demonstrate that introgressive hybridization has occurred multiple times, at least for mitochondria, despite differences among, and stability within, morphological characters that distinguish local populations. Thus, hybridization and introgression could have been key processes in the evolutionary diversification of Ohomopterus. PMID- 11903898 TI - Source population of dispersing rock-wallabies (Petrogale lateralis) identified by assignment tests on multilocus genotypic data. AB - The ability to confidently identify or exclude a population as the source of an individual has numerous powerful applications in molecular ecology. Several alternative assignment methods have recently been developed and are yet to be fully evaluated with empirical data. In this study we tested the efficacy of different assignment methods by using a translocated rock-wallaby (Petrogale lateralis) population, of known provenance. Specimens from the translocated population (n = 43), its known source population (n = 30) and four other nearby populations (n = 19-32) were genotyped for 11 polymorphic microsatellite loci. The results identified Bayesian clustering, frequency and Bayesian methods as the most consistent and accurate, correctly assigning 93-100% of individuals up to a significance threshold of P = 0.01. Performance was variable among the distance based methods, with the Cavalli-Sforza and Edwards chord distance performing best, whereas Goldstein et al.'s (deltamu)2 consistently performed poorly. Using Bayesian clustering, frequency and Bayesian methods we then attempted to determine the source of rock-wallabies which have recently recolonized an outcrop (Gardners) 8 km from the nearest rock-wallaby population. Results indicate that the population at Gardners originated via a recent dispersal event from the eastern end of Mt. Caroline. This is only the second published record of dispersal by rock-wallabies between habitat patches and is the longest movement recorded to date. Molecular techniques and methods of analysis are now available to allow detailed studies of dispersal in rock-wallabies and should also be possible for many other taxa. PMID- 11903897 TI - Fine-scale genetic structure, estuarine colonization and incipient speciation in the marine silverside fish Odontesthes argentinensis. AB - The identification of incipient ecological species represents an opportunity to investigate current evolutionary process where adaptive divergence and reproductive isolation are associated. In this study we analysed the genetic structure of marine and estuarine populations of the silverside fish Odontesthes argentinensis using nine microsatellite loci and 396 bp of the mitochondrial DNA (mtDNA) control region. Our main objective was to investigate the relationship among estuarine colonization, divergent selection and speciation in silversides. Significant genetic structure was detected among all marine and estuarine populations. Despite the low phylogeographic structure in mtDNA haplotypes, there was clear signal of local radiations of haplotypes in more ancient populations. Divergence among marine populations was interpreted as a combined result of homing behaviour, isolation by distance and drift. On the other hand, ecological shifts due to the colonization of estuarine habitats seem to have promoted rapid adaptive divergence and reproductive isolation in estuarine populations, which were considered as incipient ecological species. This conclusion is supported by the existence of a set of environmental factors required for successful reproduction of estuarine ecotypes. The pattern of genetic structure indicates that phenotypic and reproductive divergence evolved in the face of potential gene flow between populations. We suggest that the 'divergence-with-gene-flow' model of speciation may account for the diversification of estuarine populations. The approach used can potentially identify 'incipient estuarine species', being relevant to the investigation of the evolutionary relationships of silversides in several coastal regions of the world. PMID- 11903899 TI - Identification of the origin of an Atlantic salmon (Salmo salar L.) population in a recently recolonized river in the Baltic Sea. AB - The founder event in a recently recolonized salmon population in the Baltic Sea (Gulf of Finland) was investigated. To identify the origin of the founders, four wild populations and two hatchery stocks were analysed using six microsatellite loci. The results of assignment tests and factorial correspondence analysis suggest that the initial recolonizers of the river Selja originated from the geographically nearest (7 km) wild population (river Kunda) but as the result of stocking activities, interbreeding between recolonizers and hatchery individuals has occurred in subsequent years. Although the hatchery releases are outnumbering the wild salmon recruitment in the Baltic Sea at present, our results suggest that the native populations may still have an important role in colonization processes of the former salmon rivers. PMID- 11903900 TI - Strategies for microsatellite isolation: a review. AB - In the last few years microsatellites have become one of the most popular molecular markers used with applications in many different fields. High polymorphism and the relative ease of scoring represent the two major features that make microsatellites of large interest for many genetic studies. The major drawback of microsatellites is that they need to be isolated de novo from species that are being examined for the first time. The aim of the present paper is to review the various methods of microsatellite isolation described in the literature with the purpose of providing useful guidelines in making appropriate choices among the large number of currently available options. In addition, we propose a fast and easy protocol which is a combination of different published methods. PMID- 11903901 TI - Microarrays in ecology and evolution: a preview. AB - Microarray technology provides a new tool with which molecular ecologists and evolutionary biologists can survey genome-wide patterns of gene expression within and among species. New analytical approaches based on analysis of variance will allow quantification of the contributions of among individual variation, genotype, sex, microenvironment, population structure, and geography to variation in gene expression. Applications of this methodology are reviewed in relation to studies of mechanisms of adaptation and divergence; delineation of developmental and physiological pathways and networks; characterization of quantitative genetic parameters at the level of transcription ('quantitative genomics'); molecular dissection of parasitism and symbiosis; and studies of the diversification of gene content. Establishment of microarray resources is neither prohibitively expensive nor technologically demanding, and a commitment to development of gene expression profiling methods for nonmodel organisms could have a tremendous impact on molecular and genetic research at the interface of organismal and population biology. PMID- 11903903 TI - Patterns of genetic and phenotypic variation in Iris haynei and I. atrofusca (Iris sect. Oncocyclus = the royal irises) along an ecogeographical gradient in Israel and the West Bank. AB - Iris haynei and I. atrofusca are two closely related narrow endemics distributed vicariously along an ecogeographical north-south gradient in Israel and the West Bank. To obtain baseline information of the taxonomic status, conservation and population history of these taxa, we investigated patterns of phenotypic variation and the partitioning of genetic variation within and among populations using dominant random amplified polymorphic DNA (RAPD) markers. Multivariate (principal components analysis) and taxonomic distance analyses based on morphometric traits from eight populations revealed no unambiguous separation into two distinct groups. Results of genetic analyses for nine populations differed only slightly when either allele- or marker-based approaches were employed. Mean within-population diversity was high (0.258 for Nei's expected heterozygosity), but there was no significant relationship between genetic diversity and either population size or latitude. Although the range-wide estimate of GST ( approximately 0.20) revealed relatively high differentiation among populations this value was inflated because of a small, but significant, component of molecular variance among regions viz. taxa ( approximately 5%). Limited long-distance dispersal capabilities in conjunction with a linearized habitat distribution are proposed to contribute to the approximate isolation by distance pattern observed. It also appears that extant populations are currently deviating from equilibrium conditions because of primary divergence of a formerly more widespread ancestral population. Given the absence of deep genetic and phenotypic subdivision among northern (I. haynei) vs. central/southern (I. atrofusca) populations, we argue for a revision of their species status. Nonetheless, we recommend conservation attention to these geographically differentiated segments as separate management units, which can be seen as an instructive example of incipient species formation. PMID- 11903902 TI - The population genetics of host specificity: genetic differentiation in dove lice (Insecta: Phthiraptera). AB - Some species of parasites occur on a wide range of hosts while others are restricted to one or a few host species. The host specificity of a parasite species is determined, in part, by its ability to disperse between host species. Dispersal limitations can be studied by exploring the genetic structure of parasite populations both within a single species of host and across multiple host species. In this study we examined the genetic structure in the mitochondrial cytochrome oxidase I (COI) gene of two genera of lice (Insecta: Phthiraptera) occurring on multiple sympatric species of doves in southern North and Central America. One genus, Columbicola, is generally less host-specific than the other, Physconelloides. For both genera we identified substantial genetic differentiation between populations of conspecific lice on different host species, generally 10-20% sequence divergence. This level of divergence is in the range of that often observed between species of these two genera. We used nested clade analysis to explore fine scale genetic structure within species of these feather lice. We found that species of Physconelloides exhibited more genetic structure, both among hosts and among geographical localities, than did species of Columbicola. In many cases, single haplotypes within species of Columbicola are distributed on multiple host species. Thus, the population genetic structure of species of Physconelloides reveals evidence of geographical differentiation on top of high host species specificity. Underlying differences in dispersal biology probably explain the differences in population genetic structure that we observed between Columbicola and Physconelloides. PMID- 11903905 TI - Chloroplast DNA diversity in Calluna vulgaris (heather) populations in Europe. AB - Variation in the chloroplast genome of Calluna vulgaris (heather), the dominant species of northwest European heath communities, was analysed using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) and microsatellites. No length polymorphisms were detected in the 100-200 base pair (bp) fragments amplified by the conserved microsatellite primers, and sequencing revealed that the repeat regions were interrupted relative to the corresponding sequence in Nicotiana tabacum. In contrast, PCR-RFLP analysis revealed high levels of haplotype diversity within populations (hS = 0.443, hT = 0.842), as well as substantial differentiation between populations (GST = 0.473). Diversity and differentiation were higher in southern Europe than in northern Europe. Interpreted in the light of data from allozyme studies and pollen core records, the results suggest that the main glacial refugia for C. vulgaris were located in southwest Europe, including northern Spain, the Pyrenees and the Massif Central region of France. There is also evidence for diffuse survival of the species at more northerly latitudes. PMID- 11903904 TI - Population structure, history and gene flow in a group of closely related land snails: genetic variation in Partula from the Society Islands of the Pacific. AB - Previous studies of Partula land snails from the Society Islands, French Polynesia, have shown that populations within species are highly differentiated in terms of their morphology, behaviour, ecology and molecular genetic variation. Despite this level of variability, differences between species are sometimes small, possibly reflecting the fact that reproductive isolation is not always complete and there exists the opportunity for genetic exchange between taxa through hybridization. The present study uses sequence data from a mitochondrial gene to further investigate genetic variation in Society Island Partula. Most populations are found in this study to be highly differentiated, but within individual species there seems to be no simple relationship either between genetic distance and geographical proximity, or between variation in mitochondria and that in allozymes or morphological characteristics. Among species there appears to be no simple correlation between degrees of reproductive isolation and genetic relatedness according to mitochondrial DNA. The results suggest that past events as well as ongoing drift and selection may have been important in affecting patterns of variation. Similarities among species at specific localities suggest that there must have been some genetic exchange in the past, although this may not necessarily reflect ongoing rates of hybridization. The discrepancy between results for different markers probably reflects the differential effects of drift and selection on mitochondrial and nuclear genes. PMID- 11903906 TI - Phylogenetic relationships among the Canary Island Steganacaridae (Acari, Oribatida) inferred from mitochondrial DNA sequence data. AB - The mite genus Steganacarus is represented in the Canary Islands by three endemic species, one recently discovered species, and several morphotypes of uncertain taxonomic position. We used a fragment of the mitochondrial cytochrome oxidase I gene to reconstruct the phylogenetic relationships among representatives of the different taxa from the three central islands of the archipelago, Tenerife, La Gomera and Gran Canaria. Sequence data were analysed by both maximum parsimony and maximum likelihood methods. The inferred phylogenetic relationships do not correlate well with current morphological taxonomy but reveal four deeply divergent and geographically coherent lineages, one each on Gran Canaria and La Gomera and two on Tenerife. No pattern of molecular differentiation was observed among different morphotypes. Possible explanations for this incongruence are suggested in relation to the ecology and biogeography of the group. A recently discovered Steganacarus species from La Gomera, morphologically quite distinct from the other Canarian Steganacarus, is clearly identified as a taxon distantly related to all the other Canarian samples. PMID- 11903907 TI - Male reproductive competition in spawning aggregations of cod (Gadus morhua, L.). AB - Reproductive competition may lead to a large skew in reproductive success among individuals. Very few studies have analysed the paternity contribution of individual males in spawning aggregations of fish species with huge census population sizes. We quantified the variance in male reproductive success in spawning aggregations of cod under experimental conditions over an entire spawning season. Male reproductive success was estimated by microsatellite-based parentage analysis of offspring produced in six separate groups of spawning cod. In total, 1340 offspring and 102 spawnings distributed across a spawning season were analysed. Our results show that multiple males contributed sperm to most spawnings but that paternity frequencies were highly skewed among males, with larger males on average siring higher proportions of offspring. It was further indicated that male reproductive success was dependent on the magnitude of the size difference between a female and a male. We discuss our results in relation to the cod mating system. Finally, we suggest that the highly skewed distribution of paternity success observed in cod may be a factor contributing to the low effective population size/census population size ratios observed in many marine organisms. PMID- 11903908 TI - Barriers to gene flow from oilseed rape (Brassica napus) into populations of Sinapis arvensis. AB - One concern over growing herbicide-tolerant crops is that herbicide-tolerance genes may be transferred into the weeds they are designed to control. Brassica napus (oilseed rape) has a number of wild relatives that cause weed problems and the most widespread of these is Sinapis arvensis (charlock). Sinapis arvensis seed was collected from 102 populations across the UK, within and outside B. napus-growing areas. These populations were tested for sexual compatibility with B. napus and it was found that none of them hybridized readily in the glasshouse. In contrast to previous studies, we have found that hybrids can be formed naturally with S. arvensis as the maternal parent. Six diverse B. napus cultivars (Capricorn, Drakkar, Falcon, Galaxy, Hobson and Regent) were tested for their compatibility with S. arvensis but no cultivar hybridized readily in the glasshouse. We were unable to detect gene transfer from B. napus to S. arvensis in the field, confirming the extremely low probability of hybridization predicted from the glasshouse work. PMID- 11903909 TI - Differential patterns of spatial divergence in microsatellite and allozyme alleles: further evidence for locus-specific selection in the acorn barnacle, Semibalanus balanoides? AB - We compared patterns of genetic structure at potentially selected (two allozyme loci) and neutral molecular markers (six microsatellite loci) in the acorn barnacle, Semibalanus balanoides from the Gulf of St. Lawrence. Our results confirmed the presence of a geographical shift in alleles MPI and GPI near the Miramichi River. In contrast, no significant patterns of population differentiation among samples located north and south of the river mouth were detected for four of six microsatellite loci. However, analysis of molecular variance (amova) at individual loci revealed that a significant proportion of the total variance in allele frequencies was partitioned among samples located north and south of the river for both the allozyme and the other two microsatellite loci. The two most common alleles at these microsatellites showed frequencies that were highly correlated (r = 0.65-0.74, P < 0.05) with those of the MPI*2 allele, perhaps because of either physical linkage or epistasis. The two allozyme loci were significantly correlated in barnacles located north of the Miramichi River (r = 0.86, P < 0.05). Overall, our results supported the hypothesis that the broad scale pattern of allozyme allelic shifts is maintained by selection. They also indicated that microsatellites may not always behave in a neutral way and must be used cautiously, especially when evidence for genetic structuring relies on only a few assayed loci. PMID- 11903910 TI - Sex-specific genetic differentiation and coalescence times: estimating sex-biased dispersal rates. AB - I derive the equilibrium values of sex-specific FST parameters, in an island model for a dioecious species with sex-biased dispersal and binomial distribution of family size before dispersal (as assumed in a Wright-Fisher population). I show that FST may take different values among males and among females whenever dispersal is a trait conditioned on gender. This has not always been recognized, because some models assumed that genes are sampled before dispersal. In particular, the ratios of sex-specific FST parameters evaluated after dispersal over FST evaluated before dispersal are simple functions of sex-specific dispersal rates. Therefore, a simple moment-based estimator of sex-specific dispersal rate is proposed. This method is based on the comparison of FST estimated before and after dispersal and assumes equilibrium between migration and drift. I evaluate this method through stochastic simulations for a range of sex-specific dispersal rates and sampling effort (sample size, number of loci scored). PMID- 11903911 TI - Data from amplified fragment length polymorphism (AFLP) markers show indication of size homoplasy and of a relationship between degree of homoplasy and fragment size. AB - We investigate the distribution of sizes of fragments obtained from the amplified fragment length polymorphism (AFLP) marker technique. We find that empirical distributions obtained in two plant species, Phaseolus lunatus and Lolium perenne, are consistent with the expected distributions obtained from analytical theory and from numerical simulations. Our results indicate that the size distribution is strongly asymmetrical, with a much higher proportion of small than large fragments, that it is not influenced by the number of selective nucleotides nor by genome size but that it may vary with genome-wide GC-content, with a higher proportion of small fragments in cases of lower GC-content when considering the standard AFLP protocol with the enzyme MseI. Results from population samples of the two plant species show that there is a negative relationship between AFLP fragment size and fragment population frequency. Monte Carlo simulations reveal that size homoplasy, arising from pulling together nonhomologous fragments of the same size, generates patterns similar to those observed in P. lunatus and L. perenne because of the asymmetry of the size distribution. We discuss the implications of these results in the context of estimating genetic diversity with AFLP markers. PMID- 11903913 TI - Propagating contractions of the circular muscle evoked by slow stretch in flat sheets of guinea-pig ileum. AB - Flat sheet preparations of guinea-pig ileum were stretched circumferentially and the propagation of circular muscle contractions along the preparation was investigated. Slow stretch, at 100 microm s-1, of a 50-mm long flat sheet of intestine, evoked circular muscle contraction orally, which propagated, without decrement, for up to 30 mm. This occurred despite circular muscle shortening being prevented, and in the absence of propulsion of contents. Thus, propagation in this flat sheet preparation could not explained on the basis of neuro mechanical interactions, as previously proposed. Irrespective of the length of preparations, contraction amplitude decreased significantly in the most aboral 10 15 mm of intestine. This was not due to descending inhibitory pathways, but was associated with interruption of ascending excitatory pathways near the aboral end. Slow waves were not detected in circular muscle cells in any preparation (n=8). Smooth muscle action potentials evoked in circular muscle cells, in the presence of tetrodotoxin (TTX, 0.6 micromol L-1), did not propagate for more than 1 mm in the longitudinal axis. Propagation of circular muscle activity, evoked by slow stretch of flat sheet preparations, reveals the presence of a mechanism other than myogenic spread or the neuro-mechanical interactions previously proposed to account for propagation; the nature of this mechanism remains to be determined. PMID- 11903914 TI - Pharmacological modulation of human gastric volumes demonstrated noninvasively using SPECT imaging. AB - Three-dimensional single-photon emission computed tomography (SPECT) imaging allows noninvasive measurement of human postprandial gastric accommodation. The aim of this study was to determine whether 99mTCO4-SPECT demonstrates effects on pre- and postprandial gastric volumes of intravenous (i.v.) erythromycin lactobionate and sublingual isosorbide dinitrate, as predicted from previous literature. Twenty volunteers received no medication (controls), while 12 were randomized to either i.v. erythromycin 2 mg kg-1 over 20 min, or 10 mg sublingual isosorbide. After a 10-min preprandial SPECT measurement, a standard 300-mL, 300 kcal liquid meal was ingested, followed by a 20-min postprandial measurement. Gastric images were reconstructed from transaxial images and total volume was measured using the Analyseeth software system. Fasting gastric volume was greater with isosorbide [223 +/- 14 (SE) mL vs. 174 +/- 9 mL, control; P < 0.05], and postprandial volume was lower with erythromycin [393 +/- 27 mL vs. 582 +/- 17 mL, control; P < 0.05]. The ratio of postprandial over fasting volume and mean difference between pre- and postprandial volumes were significantly lower in both drug groups compared to controls. We conclude that 99mTCO4-SPECT imaging is able to semiquantitatively demonstrate pharmacological modulation of fasting gastric volume and postprandial accommodation in humans. PMID- 11903915 TI - Intestinal motor stimulation by the 5-HT4 receptor agonist ML10302: differential involvement of tachykininergic pathways in the canine small bowel and colon. AB - 5-Hydroxytryptamine (5-HT)4 receptor agonists stimulate gut motility through cholinergic pathways, although there are data suggesting that noncholinergic (tachykininergic) excitatory pathways may also be involved. Differences may exist between the small bowel and colon. Our aims were: (i) to compare the prokinetic effect exerted by the 5-HT4 receptor agonist ML10302 in the canine small bowel and colon in vivo; and (ii) to investigate the role of tachykininergic pathways in mediating this response. In fasting, conscious dogs, chronically fitted with electrodes and strain-gauge force transducers along the small bowel and colon, intravenous injection of ML10302 (35 microg kg-1) immediately stimulated spike activity and significantly increased propagated myoelectrical events at both intestinal levels. In the small bowel, the effects of ML10302 were unchanged by previous administration of the selective NK1 tachykinin receptor antagonist SR140333, the NK2 tachykinin receptor antagonist SR48968, or the NK3 tachykinin receptor antagonist SR142801. In the colon, all tachykinin receptor antagonists significantly inhibited stimulation of spike and mechanical activity by ML10302, without affecting ML10302-induced propagated myoelectrical events. Atropine (100 microg kg-1 i.v.) suppressed the stimulatory effect of ML10302 at both intestinal levels. In conclusion, the 5-HT4 receptor agonist ML10302 induced significant prokinesia both in the small bowel and colon through activation of cholinergic pathways. Tachykininergic pathways are not involved in the ML10302-induced prokinesia in the small bowel, but they play an important role in mediating the colonic motor response to ML10302. PMID- 11903916 TI - Relaxation of canine gallbladder to nerve stimulation involves adrenergic and non adrenergic non-cholinergic mechanisms. AB - Electrical field stimulation (EFS) of dog gallbladder strips induced a frequency dependent contractile response followed by an off-relaxation that was turned into a pure inhibitory response after atropine pretreatment. Guanethidine reduced the atropine-induced relaxing responses, so an adrenergic mechanism can partially account for the nerve-mediated gallbladder relaxation. However, guanethidine pretreatment also revealed a nonadrenergic noncholinergic (NANC) relaxation induced by EFS, which was frequency independent. NANC relaxations were reduced by L-arginine methyl ester (L-NAME, 100 micromol L-1), a nitric oxide synthase inhibitor (D-p-Cl-Phe6, Leul7; 10 micromol L-1), a vasoactive intestinal peptide (VIP) receptor antagonist, and an inhibitor of haem oxygenase, (copper protoporphyrin IX; CuPP-IX; 10 micromol L-1), suggesting that nitric oxide (NO), VIP and carbon monoxide (CO), respectively, are released in response to EFS. Immunoreactivities for haem oxygenase-2 (HO-2) and VIP, and histochemical staining for NADPH diaphorase were observed in nerve cell bodies and fibres, demonstrating the presence of CO, VIP and NO as putative NANC neurotransmitters in dog gallbladder. These data support the hypothesis that NO, VIP and CO contribute to NANC relaxation of the canine gallbladder. PMID- 11903917 TI - Validation of a stable isotope gastric emptying test for normal, accelerated or delayed gastric emptying. AB - To validate a 13C-Spirulina platensis breath test for measurement of accelerated or delayed gastric emptying, we measured gastric emptying of egg containing 13C S. platensis and 99mTc-sulphur colloid by breath 13 CO2 every 15 min over 3 h and scintigraphy every 15-30 min over 5 h in 57 healthy volunteers. Thirty-three received no treatment, 10 received erythromycin, and 14 atropine. A generalized linear regression model predicted half-emptying time by scintigraphy (t1/2S) from breath 13CO2 (t1/2B) data. Accuracy was assessed by standard deviation (SD) of differences between t1/2S and t1/2B and by receiver operating characteristic (ROC) curves. Regression models using breath samples at baseline, and 45, 90, 105 and 120 min, predicted t1/2B (mean +/- SD) at 118 +/- 59 min, similar to t1/2S (118 +/- 67 min). Correlation between t1/2B and t1/2S was significant (r=0.88; P < 0.0001). Differences between t1/2S and t1/2B were: 18-19.2 min for t1/2 < 70 150 min, and 68.3 min for t1/2 > 150 min. Breath test detected abnormal emptying with a sensitivity of 86% and specificity of 80%. Thus, the 13C-S. platensis test measures gastric emptying t1/2 for solids, which is accelerated or delayed to mimic a range of conditions from dumping syndrome to severe gastroparesis, with high sensitivity and specificity. Additional breath samples are needed to increase sensitivity in detecting accelerated gastric emptying. PMID- 11903918 TI - The relationship between propagated contractions and pseudoaffective changes in blood pressure in response to intestinal distension. AB - We investigated the relationship between changes in small intestinal motility and changes in blood pressure and heart rate in response to intestinal distension. Rats were maintained under stable anaesthesia with alpha-chloralose, and jejunal motility, blood pressure and heart rate were recorded. Pressure changes during propagated contractions of the circular muscle were recorded in the jejunum when the intraluminal pressure was maintained at 10 mmHg. Raising the pressure in 10 mmHg increments from 10 mmHg to 40 mmHg increased the frequency of propagated contractions from 0.30 +/- 0.06 min-1 (mean +/- SEM) to 1.29 +/- 0.09 per min. In contrast, amplitudes of contractions above baseline pressure decreased from 19.5 +/- 0.6 mmHg to 7.8 +/- 0.5 mmHg. Simultaneously, blood pressure and heart rate were both increased. Pretreatment of rats with capsaicin, or severing the mesenteric nerves acutely, prevented these cardiovascular responses, but did not influence the changes in propagated activity caused by distension. Propagated contractions were blocked by hexamethonium (10 mg kg-1, intravenously [i.v.]) and by local application of 2% lidocaine, but propulsion was unchanged by hyoscine (1 mg kg-1, i.v.). Phentolamine (1 mg kg-1, i.v.) increased the frequency of propagated contractions. The methods described in this work allow the effects of drugs on intrinsic intestinal reflexes to be distinguished from their effects on extra-intestinal, pseudoaffective reflexes. In addition, unlike other experiments using anaesthetized rats, blood pressure increased in response to distension, as it does in mammals that are not anaesthetized. The experiments demonstrate that the neural pathways for propagated contractions that rely on intrinsic nerve circuits, including intrinsic primary afferent neurones, and the neural pathways for extrinsic reflexes that signal pain or discomfort in the intestine, which involve capsaicin-sensitive spinal afferent neurones, are independent. PMID- 11903919 TI - Diurnal variation of abdominal motor responses to colorectal distension and plasma cortisol levels in rats. AB - Most patients with functional bowel disorders complain of daytime symptoms while they remain asymptomatic at night. As symptoms are associated with heightened visceral sensitivity, we hypothesized that circadian fluctuations of the visceral sensory function occur. At four different timepoints (06.00, 12.00, 18.00 and 24.00 h), colorectal distensions (CRD) were performed in fasting conscious male Lewis rats using a balloon catheter and a barostat device. The abdominal wall contractions (behavioural pain response) were assessed during colorectal distension by abdominal wall electromyography (EMG). Plasma levels for endogenous cortisol were determined simultaneously at these timepoints. EMG responses to CRD were significantly (P < 0.05) higher at midnight and in the early morning. Plasma cortisol levels peaked in the evening. In night-active Lewis rats, the behavioural pain response to noxious visceral stimulation is augmented at night and fluctuations of visceral sensitivity are accompanied by circadian changes of plasma concentrations of endogenous cortisol. We conclude that there are marked circadian fluctuations in visceral sensory functions. Thresholds are low during time periods of normal behavioural activity. These findings suggest that fluctuation of the sensory functions may be linked to the circadian variability of symptoms in patients with functional GI disorders. PMID- 11903920 TI - Altered periodic rectal motor activity: a mechanism for slow transit constipation. AB - The pathophysiology of slow transit constipation is poorly understood. Both decreased and increased distal colonic motility have been reported. In healthy humans, a 3 cycles per minute (cpm), periodic rectal motor activity (PRMA) has been described. Our aim was to investigate the characteristics of PRMA and to assess its role in the pathogenesis of constipation. A six-sensor solid-state probe was placed with the tip sensor in the mid-transverse colon, without sedation, and prolonged colonic motility was recorded in nine patients with slow transit constipation (1M, 8F) and in 11 healthy subjects (3M, 8F). Subjects were free to ambulate. We examined the frequency, nocturnal vs. diurnal variation, and characteristics of PRMA, and its relationship to proximal colonic motility. All subjects showed PRMA. The rhythm was similar (2.5-4 cpm) in both groups. However, constipated patients exhibited a greater (P < 0.001) number of PRMA cycles than controls. The duration of each cycle and amplitude of pressure waves during PRMA were also greater (P < 0.05) at night in patients compared with controls. In patients, 40% of PRMA cycles were associated with a proximal colonic motor event compared with 81% in controls (P < 0.02). The area under the curve of all colonic pressure waves and incidence of specialized propagating pressure waves was lower (P < 0.05) in patients during daytime. When compared with controls, constipated patients exhibited reduced daytime colonic pressure waves and a higher frequency of PRMA. Most of the PRMA was unrelated to proximal colonic activity in constipated patients in contrast with findings in control patients. In addition to decreased colonic motility, this excessive and unco-ordinated phasic rectal activity may further impede stool transport and contribute to the pathogenesis of slow transit constipation. PMID- 11903921 TI - The tripeptide feG reduces endotoxin-provoked perturbation of intestinal motility and inflammation. AB - Lipopolysaccharide (LPS)-induced intestinal endotoxaemia perturbs motility and causes activation and influx of inflammatory cells into the muscle tissue. Because rat submandibular gland peptide T (SGP-T; Thr-Asp-Ile-Phe-Glu-Gly-Gly), its carboxyl-terminal fragment tripeptide, FEG (Phe-Glu-Gly) and its D-isomeric analogue, feG, modulate intestinal anaphylactic reactions, we examined whether these peptides also modulate LPS-induced intestinal endotoxaemia in conscious rats. The disruption of the fasting pattern of intestinal MMCs (migrating motor complexes), induced by intravenous LPS (20 microg kg-1) injection, was prevented by all three peptides. The extravasation of leucocytes into the peritoneal cavity and increased expression of the activation marker CD18 on mesenteric tissue leucocytes (18 h after intraperitoneal injection of LPS) were reduced by orally administered feG, which also significantly decreased the number of intestinal tissue leucocytes expressing the integrin CD18. We conclude that feG attenuates both the immediate (intestinal motility) and late ( approximately 18 h) inflammatory reactions provoked by endotoxaemia. PMID- 11903922 TI - Biphasic alterations in gastrointestinal transit following endotoxaemia in mice. AB - Lipopolysaccharide (LPS)-induced alterations of gastrointestinal transit were studied in mice using activated charcoal. LPS (10 mg kg-1) induced biphasic alterations of intestinal transit. Increase (acceleration phase) and delay (lag phase) in gastrointestinal transit were observed at 90 and 480 min after LPS injection, respectively. LPS administration induced significant increases in tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta and nitrate levels in blood serum with maximal levels observed at 1.5, 4, and 8 h after LPS administration, respectively. The effects of recombinant human lzactoferrin (rhLF) on LPS- induced alteration of gastrointestinal transit, and production of TNF-alpha, IL-1beta and nitrate were also studied. Animals were pretreated with rhLF 24 hours before intraperitoneal administration of LPS. RhLF significantly increased gastrointestinal transit during the lag phase. In addition, rhLF decreased the level of TNF-alpha in endotoxaemic animals. The levels of IL-1beta and nitrate were not significantly changed by rhLF. In conclusion, the effect of LPS on gastrointestinal transit is biphasic and the mechanism controlling the second phase most likely depends on TNF-alpha production, while the first phase most likely does not depend on TNF-alpha. On the other hand, it may be regulated by IL-1beta and nitric oxide production. PMID- 11903925 TI - Cellular stress and injury responses in the brains of adult Vietnamese patients with fatal Plasmodium falciparum malaria. AB - Immunohistochemical techniques have been used to investigate specific patterns of potentially reversible cellular injury, DNA damage, and apoptosis in the brainstems of Vietnamese patients who died of severe Plasmodium falciparum malaria. The degree and pattern of neuronal and glial stress responses were compared between patients with cerebral and non-cerebral malaria (CM), and appropriate non-malaria infected controls. The following markers were examined: (i) heat shock protein 70 (HSP70), for reversible injury; (ii) heme oxygenase-1, for oxidative stress; (iii & iv) two DNA-repair proteins, poly(ADP) ribose polymerase (PARP) and DNA-dependent protein kinase catalytic subunit; (v) poly(ADP) ribose, an end-product of PARP activity; and (vi) caspase-3-active, for apoptosis. Stress responses were found in a range of cell types as reflected by the widespread expression of HSP70. Oxidative stress predominated in the vicinity of vessels and haemorrhages. Some degree of DNA damage was found in the majority of malaria patients, but the distribution and frequency of the damage was much less than that observed in controls with irreversible neuronal injury. Similarly, caspase-3-active expression, as a measure of apoptosis, was no higher in the majority of malaria patients than the negative control cases, although 40% of CM cases expressed caspase-3-active in a small number of neurones of the pontine nuclei or within swollen axons of the pontocerebellar and corticospinal tracts. In conclusion, cells within the brainstem of all patients who died from severe malaria showed staining patterns indicative of considerable stress response and reversible neuronal injury. There was no evidence for a specific pattern of widespread irreversible cell damage in those patients with cerebral malaria. PMID- 11903926 TI - Expression of peripherin in the brain of macaques (Macaca mulatta and Macaca fascicularis) occurs in astrocytes rather than neurones and is associated with encephalitis. AB - Peripherin is a member of the type III intermediate filament family, expressed in neurones of the peripheral nervous system of many species and in a discrete subpopulation of neurones of the central nervous system (CNS) during early development in rodents. Previous studies on rats have shown that peripherin immunoreactivity increased significantly in cell bodies of spinal motor neurones following axonal injury. Our study examined the expression of peripherin in the cerebrum of normal macaques (Macaca mulatta and Macaca fascicularis) and those with encephalitis of viral (simian immunodeficiency virus and simian virus 40) or autoimmune (experimental allergic encephalomyelitis) aetiology. Immunohistochemistry, immunoelectronmicroscopy, immunofluorescence and confocal microscopy were performed on tissue sections using antibodies against cell specific markers and peripherin. Peripherin-positive cells were absent in the cerebrum of normal macaques of all ages examined, whereas animals with encephalitis had peripherin-positive cells associated with inflammatory infiltrates. Further evaluation revealed that these peripherin-positive cells were not neurones, but were predominantly astrocytes expressing glial fibrillary acidic protein. Our study suggests that peripherin is not neurone-specific in the CNS of macaques; peripherin is expressed in astrocytes of animals with encephalitis. PMID- 11903927 TI - Deafferentation-induced abnormal neurofilament phosphorylation in red nucleus neurones. AB - Hippocampal deafferentation has been proposed as a pathogenetic mechanism for neurofibrillary tangle (NFT) formation in human mesolimbocortical dementia. We previously developed a rodent model of hippocampal deafferentation involving bilateral destructive lesions of the ventrotegmental area (VTA), septum of the medial forebrain and entorhinal cortex combined with pharmacological inhibition of serotonin 5-HT2 and dopamine D1 receptors. Unexpectedly, we observed an alteration in phosphorylated neurofilament protein immunoreactivity and argyrophilia in magnocellular neurones of the red nucleus. Here, we determined the neuroanatomical, pharmacological and temporal requirements for this effect on red nucleus neurones. We found that abnormal phosphorylation and argyrophilia were critically dependent on bilateral destruction of the VTA and antagonism of 5 HT2 receptors. Although extensive neurofilament hyperphosphorylation and argyrophilia were observed in red nucleus magnocellular neurones within nine days of treatment, no NFTs were formed and these effects were transitory. Resolution of these cytoskeletal abnormalities was accompanied by increased expression of the calcium binding protein, parvalbumin, suggesting that alterations in intraneuronal calcium levels may modify the deafferentation response. PMID- 11903928 TI - The glioma-associated gangliosides 3'-isoLM1, GD3 and GM2 show selective area expression in human glioblastoma xenografts in nude rat brains. AB - This work describes the in vivo expression and distribution of glioma-associated gangliosides (GD3, GM2, 3'-isoLM1) in a novel human brain tumour nude rat xenograft model. In this model, the tumours, which are established directly from human glioblastoma biopsies, show extensive infiltrative growth within the rat brain. This model therefore provides an opportunity to study ganglioside expression not only within the macroscopic tumour, but also in brain areas with tumour cell infiltration. The ganglioside expression was studied by confocal microscopy of immunostained brain sections using antiganglioside monoclonal antibodies. Xenografts from four human glioblastoma multiformes were established in rats and the brains removed after 3-4 months. Ganglioside GD3 was expressed in the tumour parenchyma while ganglioside 3'-isoLM1 was more abundantly expressed in the periphery of the tumour associated with areas of tumour cell invasion. GM2 expression was only seen in one tumour, where it was located within the main tumour mass. Double staining with a pan antihuman monoclonal antibody (3B4) and the antiganglioside monoclonal antibodies confirmed that the ganglioside expression was associated with tumour cells. This work supports the concept of different biological roles for individual gangliosides and indicates that antibodies or ligands directed against GD3 and 3'-isoLM1 might be complementary when applied in the treatment of human glioblastomas. PMID- 11903929 TI - Expression of P-selectin and intercellular adhesion molecule-1 in human brain after focal infarction or cardiac arrest. AB - Data from experimental studies indicate that acute inflammation contributes to ischaemic brain damage. Tethering of neutrophils to brain endothelium is mediated by selectins, and subsequent adhesion and migration by endothelial intercellular adhesion molecule-1 (ICAM-1) and neutrophil CD18. In experimental studies of ischaemia-reperfusion injury, brain damage has been ameliorated by administration of antibodies to these adhesion molecules. We studied the expression of P selectin and ICAM-1 in sections of brain from patients who had experienced cardiac arrest or focal brain infarction, and who died 3.5 h to 9 days later. Endothelial immunopositivity for both adhesion molecules was maximal at about 2-3 days then declined. Between 1 day and 3 days, P-selectin was also detected on platelets in blood vessels within infarcted tissue. Within infarcts, but not sections of brain from cardiac arrest patients, P-selectin and ICAM-1 were again detectable at 1 week, when hyperplastic endothelial cells were labelled in capillaries in and immediately adjacent to the infarcted tissue. The finding that P-selectin and ICAM-1 are upregulated within focally infarcted brain tissue supports the concept that blocking neutrophil adhesion may be of benefit in treating atherothrombotic strokes in man. PMID- 11903930 TI - Immunoreactivity for Bcl-2 and C-Jun/AP1 in hippocampal corpora amylacea after ischaemia in humans. AB - Corpora amylacea (CAm) are regarded as a hallmark of brain ageing, but little is known about their role in normal and pathological circumstances. CAm contain, in addition to glucose polymers, ageing-, stress- and proinflammatory proteins. In view of their almost universal occurrence and their cumulation with time, formation of CAm may represent a basic mechanism for the management of metabolic degradation products. In this context, we studied samples from post-mortem cases with repetitive brain hypoxic episodes in the past history. We investigated, by immunohistochemistry, the presence of Bcl-2, c-jun and bax in CAm. CAm showed immunoreactivity for the mitochondrial membrane associated protein Bcl-2, and for the major component of activator protein 1 transcriptional factor c-Jun. We found higher numbers of CAm in the hippocampus and the dentate gyrus in cases with repetitive cerebral hypoxia than in controls. We conclude that: (1) the presence of C-Jun and Bcl-2 within the glucose polymer mass of CAm may be related to mitochondrial damage and/or a transient overload of proteolytic systems during cellular injury; and (2) repetitive cellular stress during life may cause the age related increase of CAm in elderly subjects. PMID- 11903931 TI - Quantitative electroencephalography spectral analysis and topographic mapping in a rat model of middle cerebral artery occlusion. AB - Electroencephalography (EEG) has a long history in clinical evaluations of cerebrovascular disease. Distinct EEG abnormalities, such as increased slow delta activity, voltage depression and epileptiform discharge, have been identified in stroke patients. However, preclinical use of EEG analysis of cerebral ischaemia is less documented. We report a new rat model of EEG topographic mapping during permanent and transient middle cerebral artery occlusion. Ten EEG electrodes were implanted on the rat skull, symmetrically covering the cortical regions of two hemispheres. Monopolar EEG recordings were acquired from each animal at multiple time points during the initial 24 h, and again once daily for 7 days. Traditional EEG examinations, quantitative EEG (qEEG) spectral analysis and topographic EEG mapping were employed for comprehensive data analyses. Several distinct spatiotemporal EEG abnormalities were identified in the ischaemic rat brain. In the ipsilateral hemisphere, pronounced increase in delta activity was observed in each recorded area within 24 h of injury. While sharp waves and spike complexes dominated the parietal region, a nearly isoelectric EEG state was seen in the temporal region. After 48 h, spontaneous, albeit incomplete, recovery of EEG activities developed in all rats. Reperfusion appeared to promote delta and alpha recovery more efficiently. The contralateral EEG changes were also recorded in two phases: an acute moderate increase in delta activities with intermittent rhythmic activities, followed by a delayed and significant increase in beta activities across the hemisphere. The similarities of rat qEEG profiles identified in this study to that of stroke patients and the application of topographic mapping broaden our research technology for preclinical experimental studies of brain injury. PMID- 11903933 TI - Optimal blood gas management during deep hypothermic paediatric cardiac surgery: alpha-stat is easy, but pH-stat may be preferable. PMID- 11903934 TI - Scaling for size: some implications for paediatric anaesthesia dosing. PMID- 11903935 TI - Extrapolation of cardiac index from analysis of the left ventricular outflow velocities in children: implication of the relationship between aortic size and body surface area. AB - BACKGROUND: It has recently been reported in critically ill patients that a linear relationship exists between cardiac index (CI) measured with thermodilution and mean aortic blood flow velocity (MAFV). This hypothesis can be validated mathematically only if the aortic area (AA index) indexed to body surface area (BSA) remains constant and if the relationship between aortic diameter (PhiAo) and BSA is nonlinear. However, several other equations have described the relationship between BSA and, respectively, PhiAo and aortic area (AA) in children. The aim of this study was to determine if the relationships calculated between BSA and aortic size in children (without left ventricular outflow tract abnormality) could validate the hypothesis that MAFV and CI are well linked linearly, leading to its use to determine CI. METHODS: Two hundred and thirty-two measurements performed in 126 children and infants were retrospectively analysed. PhiAo was measured in the long axis view at the annulus using two-dimensional mode echocardiography with a 5-MHz transducer. Various linear and nonlinear relationships between BSA and, respectively, PhiAo, PhiAoindex, AA and AAindex were determined based on a nonlinear regression method with a model as follows: y=a(xc) + b. The comparisons between regressions were conducted based on the estimation error. RESULTS: The relationships between PhiAo and BSA appeared nonlinear and was well described by: PhiAo=2.96(BSA1/4) - 1.31 with a non-zero y-intercept and PhiAo=1.64(BSA1/2) with a zero y-intercept. In contrast, the relationships between AA and BSA were linear. The AAindex was not linked to BSA and can be considered as constant. The coefficient a of the equation appeared similar to those obtained mathematically with the relationship previously described between MAFV and CI. CONCLUSIONS: The hypothesis that CI can be extrapolated to the measurement of MAFV appears valid as regards the relationships calculated between aortic size and BSA in children without left ventricular outflow tract abnormality. PMID- 11903936 TI - Difficult tracheal intubation induced by maxillary distraction devices in craniosynostosis syndromes. AB - BACKGROUND: Difficult intubation occurred during anaesthesia for removal of maxillary distraction devices in five of seven children with syndromal craniosynostoses (four with Apert, two with Pfeiffer and one with Crouzon syndrome). METHODS: Intubation was assessed in terms of laryngeal view and an established intubation difficulty score and had been straightforward before device insertion. Difficulty was induced by trismus due to device insertion and by increased maxillary prominence. This was compounded by preexisting mandibular hypoplasia. Cephalometric analysis, with each child acting as their own control, demonstrated anterior displacement of the maxilla and increased maxillary vertical height, as well as increased protuberance of the maxillary incisors. RESULTS: All five difficult tracheal intubations were associated with preoperative Mallampati scores of 3 or 4 and the nine straightforward intubations with scores of 1 or 2. Maximal interincisor distance was less than the lower 95% confidence limit for age in all five children who were difficult to intubate at the time of device removal. No child had a failed intubation, but all had significantly increased intubation difficulty. CONCLUSIONS: In view of the risks of trauma, hypoxia and aspiration associated with difficult direct laryngoscopy, we recommend elective fibreoptic intubation at anaesthesia for removal of maxillary distraction osteogenesis devices in these children. PMID- 11903937 TI - Comparison of fast versus slow rewarming following acute moderate hypothermia in rats. AB - BACKGROUND: The aim of this study was to compare the biochemical and physiological responses of fast vs. slow rewarming from moderate hypothermia in anaesthetized rats. METHODS: Anaesthetized rats were surface cooled to 28 degrees C, for 20 min, then rewarmed either quickly over 30 min or slowly over 120 min with monitoring of vital signs, systemic vascular resistance (SVR), cardiac output, biochemical changes and activity for 31 days. RESULTS: At hypothermia, cardiac output decreased to 77 +/- 38 ml x min(-1) and lactate increased to 4.62 +/- 4.73 mmol x l(-)1. Fast rewarming caused an abrupt increase in cardiac output (270 +/- 24 ml x min(-1)) and a sharp drop in SVR (325.6 +/- 23.3 dyne x s(-1) x cm(-5)), compared with a smoother course with cardiac output (142 +/- 18 ml x min(-1), P < 0.01) and SVR (662.8 +/- 41.0 dyne x s(-1) x cm(-5), P < 0.01), measured during slow rewarming. Lactate failed to return to normal values (upon returning to normothermia) (2.5 +/- 0.75 mmol x l(-1)) only in the fast rewarming group. In both groups, activity in the open field was not different from control rats. CONCLUSIONS: In rats, moderate hypothermia for 20 min does not appear to cause lasting biochemical or behavioural consequences, whether rewarming lasted over 30 or 120 min. However, there was a greater early change in cardiac output and heart rate, due to systemic vasodilatation in the fast rewarming animals. These acute changes may have consequences in patients with compromised cardiovascular reserves. PMID- 11903938 TI - Management of general anaesthesia in infants and children with a history of idiopathic pulmonary haemorrhage. AB - BACKGROUND: Idiopathic pulmonary haemorrhage in infants is a rare disorder that is endemic to metropolitan Cleveland, Ohio. Since 1993, 32 infants with this disorder were diagnosed and treated at our institution, one of them after developing pulmonary haemorrhage during induction of anaesthesia. Of this population, five patients have undergone a total of 10 general anaesthetics at some time after the initial diagnosis of pulmonary haemorrhage. METHODS: We performed a retrospective chart review of these cases to identify whether any risk factors for anaesthesia-related morbidity were present, to review the anaesthetic technique and to identify morbidity related to residual underlying pulmonary disease. RESULTS: No patients experienced any anaesthesia related complication nor any perioperative respiratory problem. CONCLUSIONS: These data may be useful to anaesthesiologists in other geographical locations since this disorder has been reported in other parts of the USA, and presumably may exist in other areas of the world. PMID- 11903939 TI - The analgesic efficacy and neuroendocrine response in paediatric patients treated with two analgesic techniques: using morphine-epidural and patient-controlled analgesia. AB - BACKGROUND: Pain treatment is one of the main concerns of paediatric anaesthesiologists. The purpose of this study was to assess and compare the quality of analgesia and stress suppression by morphine when used [epidural (single shot) (EP) or with intravenous (i.v.) for patient-controlled analgesia (PCA) in children]. METHODS: Forty-four children, aged 5-15 years, and who were undergoing major genitourinary or lower abdominal surgery with a standardized general anaesthesia technique, were included in this study. In the EP group (n=24) 0.1 mg x kg(-1) morphine in 0.2 ml x kg(-1) saline were given epidurally at the L3-4 level and in the PCA group (n=20) 0.1 mg x kg-1 morphine was given i.v. immediately after intubation. Postoperative PCA bolus doses were 0.5 mg for patients weighing less than 20 kg, 1 mg for children weighing 20-30 kg and 1.5 mg for children weighing 30-40 kg. Blood samples were withdrawn following induction and at 1, 8, 12 and 24 h after morphine administration for measurement of blood glucose, insulin, cortisol and morphine levels. Patients were observed for 24 h postoperatively; heart rate, systolic blood pressure, respiratory rate, FACES pain scores, sedation scores and complications were recorded. RESULTS: The PCA group received 0.56 +/- 0.33 mg x kg(-1) x day(-1) morphine. The FACES pain scores, sedation scores, cortisol, blood glucose and insulin levels were similar in both groups. Haemodynamic and respiratory evaluations and cortisol levels were stable but blood glucose and insulin changes at certain time periods were significant (P < 0.05). Serum morphine levels and incidence of vomiting were different between groups (P < 0.05). Serum morphine levels were similar at the first hour. CONCLUSIONS: Both techniques provided sufficient pain relief and attenuated the hormonal response without life-threatening complications. PMID- 11903940 TI - Ilioinguinal and iliohypogastric nerve block revisited: single shot versus double shot technique for hernia repair in children. AB - BACKGROUND: We attempted to determine the efficacy of a one plane ilioinguinal and iliohypogastric nerve block with a single shot and double shot techniques. METHODS: In a randomized single blind study, 90 children, aged 2-12 years, received a single shot (SS) or a double shot (DS) technique for ilioinguinal and iliohypogastric (IG-IH) nerve block for inguinal hernia repair. In the SS group, 0.25 ml x kg(-1) of 0.25% bupivacaine was given one fingerbreadth medial to the anterior superior iliac spine under the external oblique aponeurosis. In the DS group, one-third of the total dose of bupivacaine was given as for the SS group. The remaining two-thirds was deposited 0.5 cm above and lateral to the mid inguinal point deep to the external oblique aponeurosis. RESULTS: The success rates of both techniques were similar, at 72%, although the presence of local anaesthetic in the inguinal canal was significantly higher with the DS technique. The incidence of femoral nerve block was 4.5% with the SS and 9% with the DS technique (P > 0.05). Parental satisfaction with postoperative pain relief was high, at 94%. CONCLUSIONS: The DS technique, while technically more difficult, does not improve the success rate of the IG-IH nerve block compared with the SS technique. PMID- 11903941 TI - A survey of parental satisfaction during parent present induction of anaesthesia for children undergoing cardiovascular surgery. AB - METHODS: To assess parental reaction and possible complications of parent present induction (PPI) for children undergoing cardiovascular surgery, the parents of 183 patients were provided with a questionnaire to complete after they had participated in PPI. Questions included: prior experience with PPI, which member of the staff initiated the plan for PPI, parental role in the process, how prepared parents felt, and overall satisfaction. RESULTS: PPI was successfully performed in the 183 patients surveyed. No parent was asked to leave the operating room because of respiratory or haemodynamic complications; 77.6% of the parents had no prior experience with PPI; however, 91.8% were aware of their role in the operating room, 94.5% were aware how their child would be anaesthetized and 96.7% felt prepared for their role and believed that this was a positive experience for themselves and their child. CONCLUSIONS: This prospective survey demonstrated a high level of parental acceptance and satisfaction for PPI in children undergoing cardiovascular surgery, with a low incidence of untoward events, despite the underlying congenital heart disease. Further work is necessary to objectively characterize anxiety levels associated with induction of anaesthesia in this group of patients and parents. PMID- 11903942 TI - Preoperative oral granisetron for the prevention of vomiting following paediatric surgery. AB - BACKGROUND: We evaluated the efficacy of granisetron, 5-hydroxytryptamine type 3 receptor antagonist, given orally, preoperatively, for the prevention of postoperative vomiting in children undergoing general anaesthesia for surgery (inguinal hernia, phimosis-circumcision). METHODS: In a randomized, double blinded manner, 100 children, ASA physical status I, aged 4-11 years, received orally placebo or granisetron at three different doses (20 microg x kg(-1), 40 microg x kg(-1), 80 microg x kg(-1)) 60 min before surgery (n=25 of each). The same standard general anaesthetic technique was used. RESULTS: The percentage of patients being emesis-free during 0-6 h after anaesthesia was 56% with placebo, 64% with graniseron 20 microg x kg(-1) (P=0.773), 88% with granisetron 40 microg x kg(-1) (P=0.027) and 92% with granisetron 80 microg x kg(-1) (P=0.01); the corresponding rate during 6-24 h after anaesthesia was 60%, 68% (P=0.768), 92% (P=0.02) and 92% (P=0.02) (P-values versus placebo). No clinically serious adverse events were observed in any of the groups. CONCLUSIONS: In summary, preoperative oral granisetron 40 microg x kg(-1) is effective for the prevention of vomiting following paediatric surgery (inguinal hernia, phimosis circumcision). Increasing the doses to 80 microg x kg(-1) provides no demonstrable additional benefit. PMID- 11903943 TI - Therapeutic applications of regional anaesthesia in paediatric-aged patients. PMID- 11903944 TI - Anaesthetic management of a patient with microvillus inclusion disease for intestinal transplantation. AB - We report the anaesthetic management of a 3-year-old-child with microvillus inclusion disease undergoing isolated small bowel transplantation. He required long-term total parenteral nutrition which was complicated with numerous episodes of catheter related sepsis. This resulted in thrombosis of the major blood vessels which critically restricted vascular access available for intravenous nutrition, becoming a life-threatening condition for the patient. Haemodynamic, respiratory parameters and urinary output were well preserved throughout the procedure. Besides a transitory increase in potassium following graft revascularization, biochemical changes were small. Anaesthetic management included comprehensive preoperative assessment, central venous angiography to depict accessibility of central and peripheral veins, assurance of additional vascular access through the intraoperative catheterization of the left renal vein, perioperative epidural analgesia and preservation of splanchnic perfusion to ensure implant viability. PMID- 11903945 TI - Perioperatively acquired methaemoglobinaemia in a preterm infant. AB - Methaemoglobinaemia is an uncommon cause of neonatal cyanosis. A case that illustrates the aetiology, diagnosis, and treatment of this condition is presented. PMID- 11903946 TI - An abnormal dentition in progeria. PMID- 11903947 TI - Does internal jugular vein cannulation in infants require a SMART needle rather than a smart anaesthetist? PMID- 11903949 TI - Oxygen saturation during transfer. PMID- 11903950 TI - Simpler version of a videolaryngoscope. PMID- 11903952 TI - Problems associated with the clinical use of proton pump inhibitors. AB - Proton pump inhibitors have proven to be important in the treatment of acid/peptic diseases. However, their therapeutic uses have extended over time, which may alter their risk:benefit ratio. Significant rebound acid hypersecretion has been demonstrated, which persists for at least two months. Its clinical significance remains unknown. Enhanced oxyntic gastritis may be responsible for the increased acid suppressive effects of proton pump inhibitors in H. pylori infected patients. It remains unclear whether either H. pylori eradication or diminishing rates of infection in the developed world will therefore decrease the efficacy of proton pump inhibitors. Problems with carcinoids in rodents have not been found in man. However, the consequences of very long-term use of proton pump inhibitors remain unknown. Similarly, the development of atrophic gastritis in H. pylori-positive patients treated with proton pump inhibitors, with the long-term concern of gastric carcinoma development, remains controversial. PMID- 11903953 TI - Genotoxic effects of the alkaloids harman and harmine assessed by comet assay and chromosome aberration test in mammalian cells in vitro. AB - Harman and harmine are beta-carboline alkaloids which are present in plants widely used in medical practice, in beverages used for religious purposes in Brazil, as well as in tobacco smoke and over cooked food. In view of the controversial results observed in the literature about the mutagenic effects of these alkaloids, we studied their cytotoxic and genotoxic effects in V79 Chinese hamster lung fibroblasts in vitro using single-cell gel assay, Comet assay, either in the presence or in absence of an exogenous metabolic activation system (S9-mix), and by the chromosome aberration test without S9-mix. Harmine was more cytotoxic than harman. Both harman and harmine increased aberrant cell frequency and induced DNA damage by the Comet assay. These results suggest that harman and harmine are genotoxic in V79 cells, probably as a consequence of their ability to induce DNA strand breaks. PMID- 11903954 TI - Differential sensitivity to 8-hydroxy-2-(di-n-propylamino)tetralin-induced corticosterone secretion and hypothermia between Sprague-Dawley rats from different breeding colonies. AB - The sensitivity of Sprague-Dawley rats from 4 different breeding colonies (ALAB, M&B, B&K, Charles River) and one breeding colony of Wistar rat (M&B) to the 5 hydroxytryptamine1A (5-HT1A) receptor stimulatory effect of 8-hydroxy-2-(di-n propylamino)tetralin hydrobromide (8-OH-DPAT) resulting in corticosterone secretion and hypothermia was compared. The dose-response curves of the increase in plasma corticosterone showed that ALAB and M&B rats were 3.5 times more sensitive to 8-OH-DPAT than B&K and Charles River rats, the Wistar rats being in between. The attenuation of the corticosterone response 24 hr after a single injection of 1 mg/kg 8-OH-DPAT was greater for the ALAB and M&B rats than for B&K, Charles River and Wistar rats. The comparison of the 8-OH-DPAT-induced hypothermia in the various rat colonies showed a similar pattern: the sensitivity of ALAB rats was about twice that of M&B, B&K and Wistar rats, Charles River rats being 9 times less sensitive. The attenuation of the response 24 hr after 1 mg/kg 8-OH-DPAT measured as the shift in dose-response showed similar shift factors (4.1 to 6.7) for all rat colonies except for the B&K rats (3.0). The hypothermic response at 0.1 mg/kg 8-OH-DPAT was significantly lower for the Charles River and B&K rats than for the ALAB rats. Similarly was the maximal attenuation of the hypothermic effect in these rats less than half of that of the ALAB rats. The possible cause of the observed differences in the response to 8-OH-DPAT between these rat colonies is discussed in terms of receptor reserves and the involvement of other transmitter systems in the responses. PMID- 11903955 TI - Nordihydroguaiaretic acid elevates osteoblastic intracellular Ca2+. AB - Nordihydroguaiaretic acid (NDGA) is widely used as a pharmacological tool to inhibit lipoxygenases; however, recent evidence suggests that it increases renal intracellular [Ca2+]i via novel mechanisms. Here the effect of NDGA on Ca2+ signaling in MG63 osteoblastic cells was explored using fura-2 as a Ca2+ indicator. NDGA (2-50 microM) increased [Ca2+]i in a concentration-dependent manner. The signal comprised an initial rise and an elevated phase over a time period of 4 min. Removing extracellular Ca2+ reduced 2-50 microM NDGA-induced signals by 62+/-2%. After incubation with 50 microM NDGA in Ca2+-free medium for several minutes, addition of 3 mM CaCl2 induced an increase in [Ca2+]i. NDGA (50 microM)-induced [Ca2+]i increases were not changed by pretreatment with 10 microM of verapamil, diltiazem, nifedipine, nimodipine and nicardipine. In Ca2+-free medium, pretreatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (1 microM) inhibited 50 microM NDGA-induced [Ca2+]i increases by 69+/-3%. Inhibition of phospholipase C with 2 microM U73122 had little effect on 50 microM NDGA-induced Ca2+ release. Several other lipoxygenase inhibitors had no effect on basal [Ca2+]i. At a concentration that did not increase basal [Ca2+]i, NDGA (1 microM) did not alter 10 microM ATP- or 1 microM thapsigargin-induced [Ca2+]i increases. Alteration of protein kinase C activity with 1 nM phorbol 12 myristate 13-acetate or 2 microM GF 109203X did not affect 50 microM NDGA-induced [Ca2+]i increases. Together, the results show that NDGA increased [Ca2+]i in osteoblasts in a lipoxygenase-independent manner, by releasing stored Ca2+ in a fashion independent of phospholipase C activity, and by causing Ca2+ influx. PMID- 11903956 TI - The effect of acute oxidative stress on the ultrastructure of the perfused rat liver. AB - Ageing and liver disease are associated with ultrastructural changes in the hepatic sinusoid. Because of the possibility that reactive oxygen species could mediate these processes, we examined the effect of acute oxidative stress on the ultrastructure of the intact liver. Rat livers were perfused ex vivo, in situ with hydrogen peroxide via the portal vein. The livers were then fixed and the ultrastructure of the liver tissue examined with transmission and scanning electron microscopy. The effects of hydrogen peroxide were largely confined to the perisinusoidal areas. The sinusoidal endothelial cells became swollen and more porous, with large gaps replacing sieve plates. The space of Disse showed an increase in volume and the density of hepatocyte projections decreased. Kupffer cell activation was noted. Little or no ultrastructural change was observed within the hepatocytes. Oxidative stress delivered via the portal vein dramatically alters the ultrastructure of the perisinusoidal regions of the liver. This process may contribute to the pathogenesis of disease and age-related changes in the liver. PMID- 11903957 TI - Effects of ketoconazole on oestrus cycle and convulsant action of pentylenetetrazol in mice. AB - The frequency of oestrus cycles in female mice was significantly reduced by the implantation of a pellet of subcutaneous ketoconazole (50 mg every 6 days). The effect was more pronounced after 22 days than after 13 days and it was probably related with the progressive reduction in steroid synthesis induced by the inhibition of key steroidogenic P450 cytochromes by the drug. In addition, the influence of ketoconazole on the incidence of seizures after the administration of intraperitoneal pentylenetetrazol was evaluated in female mice. Pentylenetetrazol produced a higher percentage of seizures during dioestrus than during oestrus. Pretreatment with ketoconazole significantly increased the percentage of animals with induced seizures in oestrus but not in dioestrus as compared to controls, probably through reduced progesterone levels. The reduced seizure threshold confirm the modulatory role exerted by progesterone on central nervous system excitability, and may be relevant in epileptic patients undergoing antifungal therapy. PMID- 11903958 TI - Aluminium phosphide exposure: implications on rat brain lipid peroxidation and antioxidant defence system. AB - We investigated the effect of aluminium phosphide exposure (10 mg/kg body weight) on lipid peroxidation and antioxidant defence system in different regions of rat brain. A significant increase in lipid peroxidation in cerebrum, cerebellum and brain stem was observed in aluminium phosphide-exposed rats, which was accompanied by a marked decrease in the activities of antioxidant enzyme, superoxide dismutase and catalase. A decline in the activity of glutathione reductase was also observed, however, no change was seen in the activity of glutathione peroxidase following aluminium phosphide administration. Decreased levels of non-protein thiols and total sulfhydryl groups were also observed after aluminium phosphide treatment. It seems evident that aluminium phosphide exposure significantly enhanced neuronal lipoperoxidative damage with concomitant alterations in the antioxidant defence status thus having serious bearing on the functional and structural status of the central nervous system. PMID- 11903959 TI - Copper, ceruloplasmin and superoxide dismutase (SOD1) in patients with Down's syndrome. AB - The aim of this study was primarily to investigate whether similar signs of copper dyshomeostasis occur in dementia with age in Down's syndrome as previously found in Alzheimer's disease. Copper was accordingly determined in plasma, ceruloplasmin concentration in serum, ceruloplasmin oxidative activity and ceruloplasmin specific oxidative activity (activity related to mass) in serum, and superoxide dismutase (SOD1) in erythrocytes in 35 (27 males, 8 females) 18-53 years old (average 37 years) patients with Down's syndrome (Down's patients) and their age- and gender-matched controls. SOD1 activity was on an average almost 50% higher in the patients than in their controls but the evidence of a causal relationship between increased SOD1 activity and Down's syndrome appears at best equivocal. Copper and ceruloplasmin levels and ceruloplasmin activities were similar in the patients and their controls. Ceruloplasmin and copper levels increased significantly with age in the patients but not in the controls. Ceruloplasmin activities or SOD1 activity did not change significantly with age, neither in the patients nor in the controls as whole groups. When SOD1 activity and ceruloplasmin activities of the oldest in the patients group (40 years or older) were compared with those of the younger patients, respectively, SOD1 activity and specific oxidative activity, but not ceruloplasmin oxidative activity were found to decrease significantly with age. The results thus suggest that development of dementia in Down's patients with age is paralleled with decrease in SOD1 activity and specific oxidative activity but not with decrease in ceruloplasmin oxidative activity itself as was also found in Alzheimer's patients. PMID- 11903960 TI - Calcium supplementation efficiently reduces lead absorption in suckling rats. AB - The effect of calcium supplementation on tissue lead was evaluated in suckling Wistar rats. Such data are not yet available in the literature. The following artificial feeding regimen was used for calcium supplementation: cow's milk by addition of 1%, 3% or 6% Ca as CaHPO(4)x2H(2)O suspension to increase the daily calcium intake about 1.4, 2 or 3 times above control values. Artificial feeding was applied during 7 hr each day for nine consecutive days (from day 6 through 15 after birth). The effect of such treatment on lead absorption and elimination was evaluated in two separate experiments: calcium supplementation during oral lead exposure (as acetate; daily dose 2 mg Pb/kg body wt.; total Pb dose 18 mg/kg body wt.) or after a single intraperitoneal lead administration (5 mg/kg body wt.). At the end of experiments, lead in tissues (liver, kidneys, brain and carcass), and essential elements (Ca, Fe, Zn, Cu) were analysed by atomic absorption spectrometry. Calcium supplementation caused a statistically significant decrease of lead in all tissues of sucklings orally exposed to lead. This decrease was dose-related being about 1.3, 1.5 and 2 times lower in groups supplemented with 1%, 3%, or 6% calcium compared to controls, respectively. Increased calcium intake had no effect on incorporated lead after parenteral lead exposure. Calcium supplementation increased carcass calcium and had no effect on trace elements in tissues, pups' general appearance and body weight gain. It is concluded that higher calcium intake might be a way of efficient reduction of lead absorption during the suckling period. PMID- 11903961 TI - Dose-dependent inhibition of the CYP2D6 catalyzed oxidation of sparteine by mepyramine in healthy volunteers. PMID- 11903962 TI - Changes in the activities of antioxidant enzymes in response to virus infection and hormone treatment. AB - Activities of enzymes involved in the detoxification of reactive oxygen species (catalase, glutathione reductase, peroxidase and superoxide dismutase (SOD)) were examined in the leaves of Phaseolus vulgaris L. var. Top Crop treated with plant hormones and infected with a non-lesion-forming isolate of white clover mosaic potexvirus (WClMV). The activities of catalase, glutathione reductase and SOD rapidly declined after infection while peroxidase activity was enhanced. These changes occurred before the rapid increase (5 days) in WClMV replication. A mild chlorosis appeared 7-10 days after inoculation but necrosis was never observed on inoculated leaves. Plants treated with dihydrozeatin, salicylic acid and jasmonic acid prior to WClMV inoculation showed elevated catalase, glutathione reductase, and peroxidase activity, while SOD activities remained the same as in water treated controls. These treatments all inhibited virus replication with enzyme activities remaining near control levels. We propose that a decline in free radical scavenging capacity may be required before a rapid increase in virus replication can take place. Treatments increasing the ability of the plant to scavenge reactive oxygen species may hinder virus replication. A possible role for reactive oxygen species as a requirement for virus replication is discussed. PMID- 11903963 TI - Isolation and characterization of an extended thioredoxin h from poplar. AB - A cDNA coding for a thioredoxin h has been isolated from a xylem/phloem poplar cDNA library by RACE-PCR. The nucleotide sequence called popTrx-h2 is homologous to other thioredoxins h isolated from plants but differs from the other thioredoxins h by presenting a 30 amino acid long N-terminus extension. A variant of this cDNA lacking the N-terminal extension was also generated by PCR. Both cDNAs have been introduced into an expression plasmid (pET-3d) and the recombinant proteins have been expressed to a high level and purified from Escherichia coli cells. Protein sequencing showed that a part of the N-terminal extension was cleaved in the E. coli cells, with the first 19 amino acids missing, suggesting the presence of a putative cleavage site in the N-terminal extension of popTrx-h2. Both recombinant proteins display unusual catalytic properties compared to other thioredoxins h characterized so far, i.e. a weak reduction by Arabidopsis thaliana NADPH-dependent thioredoxin reductase, and a weak activation of the chloroplastic NADP-malate dehydrogenase, a non physiological target enzyme. Northern blot experiments indicate that the transcripts of popTrx-h2 are present in leaves and roots, albeit at a lower level compared to the earlier characterized popTrx-h1. PMID- 11903964 TI - Activation of pyrophosphate:fructose-6-phosphate 1-phosphotransferase by fructose 2,6-bisphosphate stimulates conversion of hexose phosphates to triose phosphates but does not influence accumulation of carbohydrates in phosphate-deficient tobacco cells. AB - The aim of this work was to investigate the contribution of fructose 2,6 bisphosphate to the regulation of carbohydrate metabolism under phosphate stress. The study exploited heterotrophic tobacco callus lines expressing a modified mammalian 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase that increased the fructose 2,6-bisphosphate content of the tissue. The phosphate status of two transgenic and one untransformed cell line was perturbed by incubation with 2 deoxyglucose, a phosphate-sequestering agent, and by growth of callus on phosphate-depleted media. 31P-NMR spectroscopy confirmed that both treatments decreased cellular levels of inorganic phosphate and phosphorylated metabolites. Despite large decreases in the amounts of phosphate esters, UDPglucose and adenylates in response to phosphate deficiency, the fructose 2,6-bisphosphate content of each line was unaffected by 2-deoxyglucose and increased during growth on phosphate-limited media. Short-term treatment of callus with 2-deoxyglucose had only minor effects on the carbohydrate status of each line, whereas long-term phosphate deficiency caused an increase in starch and a decrease in soluble sugar content in both transgenic and control lines. There were no consistent differences between the three callus lines in metabolism of [U-14C]glucose in response to incubation with 2-deoxyglucose. In contrast, there was a decrease in partitioning of label into glycolytic products (particularly organic acids) in untransformed callus during growth on phosphate-depleted medium. This decrease was greatly attenuated in the transgenic lines with increased fructose 2,6 bisphosphate content. This suggests that the conversion of hexose phosphates to triose phosphates is constrained under phosphate-deficient conditions, and that this restriction can be relieved by activation of pyrophosphate:fructose-6 phosphate 1-phosphotransferase. However, since the transgenic and control lines did not differ in the extent to which the carbohydrate content changed in response to growth on phosphate-depleted media, it is concluded that an increase in flux through pyrophosphate:fructose-6-phosphate 1-phosphotransferase is not a major component of the metabolic response of heterotrophic tobacco cells to phosphate deficiency. PMID- 11903965 TI - Cell wall modifications of bean (Phaseolus vulgaris) cell suspensions during habituation and dehabituation to dichlobenil. AB - Bean (Phaseolus vulgaris L.) cell suspensions were adapted for growth in 12 &mgr;M dichlobenil (2,6-dichlorobenzonitrile or DCB) by a stepwise increase in the concentration of the inhibitor in each subculture. Non-tolerant suspensions (I50 = 0.3 &mgr;M) gave rise to single cells or small clusters while tolerant cell suspensions (I50 = 30 &mgr;M) grown in DCB formed large clusters. The cells in these clusters were surrounded by a thick and irregular cell wall with a lamellate structure and lacking a differentiated middle lamella. Analysis of habituated cell walls by Fourier transform infrared spectroscopy and cell wall fractionation revealed: (1) a reduced amount of cellulose and hemicelluloses, mainly xyloglucan (2) qualitative and quantitative differences in pectin levels, and (3) a non-crystalline and soluble beta-1,4-glucan. When tolerant cells were returned to medium lacking DCB, the size of the cell clusters was reduced; the middle lamella was only partly formed, and the composition of the cell wall gradually reverted to that obtained with non-tolerant cells. However, dehabituated cells (I50 = 12 &mgr;M) were 40-fold more tolerant to DCB than non tolerant cells and were only 2.5-fold more sensitive than tolerant cells. PMID- 11903966 TI - Impact of four years exposure to different levels of ozone, phosphorus and drought on chlorophyll, mineral nutrients, and stem volume of Norway spruce, Picea abies. AB - Saplings of one clone of Norway spruce, Picea abies (L.) Karst, were planted in 120 l pots in 1991 and exposed to three levels of ozone, two levels of phosphorus and two levels of water supply in 42 open-top chambers (OTCs), during 1992-1996. The effects of pots and OTCs were also tested. Nutrient concentrations of the needles were not affected by ozone, while the low phosphorus supply (LP) and drought stress (D) treatments had significant effects on several mineral nutrients, e.g. phosphorus, calcium, magnesium, manganese, sulphur and boron. Ozone reduced the chlorophyll concentration in the 2- and 3-year-old needles in 1994 and 1995. The highest ozone concentration reduced the stem volumes (- 8%), as well as the stem lengths (- 5%), of the saplings in 1993 and 1994, after two and three years of exposure. After the fourth growing season this ozone-induced reduction in stem volume disappeared which might be caused by pot limitation. LP supply and D both caused large decreases in the stem volume and length. The needles from LP treatment had as high P concentration as 1.2-1.5 mg g-1, implying a need for increasing the critical value for phosphorus. The OTC enclosure stimulated the stem volume growth significantly compared to saplings growing in ambient plots. This was suggested to be attributed to the slightly higher temperature in the OTCs. The overall result is that ozone in southern Sweden is likely to have negative effects on Norway spruce trees, although much less than other environmental factors, e.g. water and phosphorus. PMID- 11903967 TI - Requirement for far-red light to maintain secondary needle extension growth in northern but not southern populations of Pinus sylvestris (Scots pine). AB - Extension growth of secondary needles is under photoperiodic control in Pinus sylvestris. To test for the effects of far-red light on maintaining this extension growth, seedlings of six populations originating from latitudes between 57 degrees and 67 degrees N were raised for 11 weeks in continuous incandescent (metal halogen) light at 300 &mgr;mol m-2 s-1 and 20 degrees C and then transferred at the same temperature to a daily regime of 8 h incandescent light (230 &mgr;mol m-2 s-1) followed by a 16 h day extension with cool white fluorescent light (40 &mgr;mol m-2 s-1, R/FR ratio 7.5) or with incandescent lamps (20 &mgr;mol m-2 s-1, R/FR ratio 2.0). For the seedlings from the three populations north of 64 degrees, needle extension growth over 42 days in the FR poor day extension treatment was lower by up to 40% than in the FR-rich day extension treatment, whereas for the seedlings from the three southern populations the needle extension growth was similar in both day extension treatments. The requirement for FR in day extensions is characteristic of 'light dominant' photoperiodic control mechanisms. It appears that P. sylvestris changes from dark-dominant night timekeeping to light-dominant day timekeeping with increasing latitude, as with the photoperiodic control of budset in Picea abies. PMID- 11903968 TI - Pectin-bound beta-galactosidase present in cell walls of carrot cells under the different calcium status. AB - Glycosyl-hydrolytic enzymes from suspension-cultured carrot (Daucus carota L. cv. Kintoki) cells grown in calcium (Ca2+)-deficient and normal liquid media were studied after extraction successively by K-phosphate (pH 7.0) and Na-acetate (pH 5.2) containing 3 M LiCl. The same activities were detected in two protein fractions from control and Ca2+-deprived cells. The specific activities of alpha galactosidase and polygalacturonase decreased under Ca2+ deprivation, but beta galactosidase activity in the buffer-soluble protein from Ca2+-deprived cells increased 1.7-fold compared to control cells. Upon ion exchange and size exclusion chromatography the fraction (Ca-Ia-I) in the buffer-soluble protein from Ca2+-deprived cells represented beta-galactosidase activity associated with a galacturonic acid-rich polysaccharide peak, whereas the corresponding fraction could hardly be detected in the buffer-soluble protein from control cells. Several of the same glycosidase activities were detected in the extract solubilized with cyclohexane-trans-1,2-diaminetetra-acetate (CDTA) from active cell walls of Ca2+-deprived cells as in the extract of control cells, but the beta-galactosidase activity was considerably reduced under Ca2+ deprivation. Following the same chromatography the fraction (CDTA-Ca-1) of beta-galactosidase activity in the extract solubilized with CDTA from active cell walls of Ca2+ deprived cells was also completely overlapping with the peak of galacturonic acid rich polysaccharide. The molecular mass of fractions Ca-Ia-I and CDTA-Ca-1 was 300 kDa, and the polysaccharides in these two fractions were composed of approximately equal amounts of rhamnosyl and galacturonosyl residues. These results suggest that the increase of beta-galactosidase in the buffer-soluble protein fraction from Ca2+-deprived cells is the result of solubilization of a part of the acidic pectic polymer-bound beta-galactosidase due to the structural changes in the cell walls that occur during Ca2+ deprivation. PMID- 11903969 TI - Differences in salt sensitivity of four deciduous tree species to soil or airborne salt. AB - Seedlings of four deciduous tree species maple (Acer pseudoplatanus), beech (Fagus sylvatica), horse chestnut (Aesculus hippocastanum) and lime (Tilia cordata) were exposed to de-icing salt (NaCl) either through the soil or applied to the above ground plant parts. A soil solution of 1.65 g l-1 NaCl was maintained from the start of the experiment in January 1999 until termination in June 1999. The main effects caused by salt treatment through the soil were a reduction in photosynthesis of up to 50% and the development of leaf chlorosis or necrosis covering up to 50% of the total leaf area for the most sensitive species (lime and beech); maple and horse chestnut were relatively tolerant. There was no significant correlation between Cl or Na concentration in leaves and the relative sensitivity of the species. Saturated salt solution was applied to bark, buds or leaf scars on two occasions three weeks apart during the winter season. This affected the timing of bud break with delays of up to eight days compared with the controls. In the most sensitive species the above ground salt treatments partly prevented bud break (beech) or reduced photosynthesis (lime). Uptake through the bark was most important for the development of stress effects, compared with uptake through the other above ground plant parts. PMID- 11903970 TI - Steady-state chlorophyll fluorescence (Fs) measurements as a tool to follow variations of net CO2 assimilation and stomatal conductance during water-stress in C3 plants. AB - Water stress experiments were performed with grapevines (Vitis vinifera L.) and other C3 plants in the field, in potted plants in the laboratory, and with detached leaves. It was found that, in all cases, the ratio of steady state chlorophyll fluorescence (Fs) normalized to dark-adapted intrinsic fluorescence (Fo) inversely correlated with non-photochemical quenching (NPQ). Also, at high irradiance, the ratio Fs/Fo was positively correlated with CO2 assimilation in air, with electron transport rate calculated from fluorescence, and with stomatal conductance, but no clear correlation was observed with qP. The significance of these relationships is discussed. The ratio Fs/Fo, measured with a portable instrument (PAM-2000) or with a remote sensing FIPAM system, provides a good method for the early detection of water stress, and may become a useful guide to irrigation requirements. PMID- 11903971 TI - Differential expression and characterization of three metallothionein-like genes in Cavendish banana (Musa acuminata). AB - Metallothioneins (MTs) are cysteine-rich polypeptides that are involved in metal detoxification and homeostasis in both prokaryotes and eukaryotes. In this study, we report the isolation and characterization of three members (MT2A, MT2B and MT3) of the MT-like gene family from ripening banana fruit and their differential expression in various banana organs and during fruit development and ripening. All members of the MT-like gene encode small cysteine-rich polypeptides of 65-79 amino acid residues. MT2A shared a high sequence similarity (54-77%) with several type-2 MTs in plants, while MT3 was highly homologous (51-61%) with type-3 MTs. The three members expressed differentially in various organs but transcripts were generally more abundant in reproductive than vegetative organs. During fruit development, the MT2A transcript was barely detectable in ovary but increased to a high level in young fruit at 20 days after shooting (DAS) and declined gradually thereafter as fruit developed. In contrast, both MT2B and MT3 expressed poorly in young fruits (20-60 DAS) and transcripts were detected only in fruits at later stages of development. As ripening progressed, expression of MT2A decreased but that of MT3 increased. Expression of MT members during ripening appeared to be differentially regulated by ethylene, whose levels were low in FG and TY fruit but surged climacteristically in MG and declined sharply as ripening advanced further. Exogenous application of ethylene at 5 ppm or higher concentrations down-regulated MT2A expression and the inhibitory effect of ethylene could be partially suppressed by the presence of norbornadiene, an inhibitor of ethylene action. Ethylene had no effect on transcript accumulation of MT2B and MT3. However, MT3 expression was greatly enhanced in response to metals such as CdSO4, CuSO4 and ZnSO4. These results suggest that increased MT3 expression may be associated with excess metal ions present in ripening fruit tissues. This study also provided evidence, for the first time, that ethylene and metals play a regulatory role in expression of MT-like genes in banana. PMID- 11903972 TI - Flavonol synthase gene expression during citrus fruit development. AB - We isolated a cDNA clone (CitFLS) encoding flavonol synthase (FLS) from the satsuma mandarin (Citrus unshiu Marc.) fruit and investigated the steady state of CitFLS RNA expression during the fruit development. The CitFLS was 1274 bp long, encoded 335 amino acid residues, and belonged to a family of 2-oxoglutarate dependent dioxygenases. The level of CitFLS transcript was higher in the young leaves than in the old leaves, and it was high at the early developmental stage and low at the mature stage in the juice sacs/segment epidermis (edible part). On the other hand, the CitFLS transcript increased in the peel during fruit maturation. These results indicated that the satsuma mandarin CitFLS was differentially regulated in the developmental stage and in a tissue-specific manner. Additionally, satsuma mandarin peel tissues produced rutin (a flavonol glycoside) from an exogenous dihydroquercetin (taxifolin), indicating the ability of these tissues to produce flavonols. PMID- 11903973 TI - Isolation and characterization of six peach cDNAs encoding key proteins in organic acid metabolism and solute accumulation: involvement in regulating peach fruit acidity. AB - As in many other fleshy fruits, the predominant organic acids in ripe peach (Prunus persica (L.) Batsch) fruit are malic and citric acids. The accumulation of these metabolites in fruit flesh is regulated during fruit development. Six peach fruit-related genes implicated in organic acid metabolism (mitochondrial citrate synthase; cytosolic NAD-dependent malate dehydrogenase, and cytosolic NADP-dependent isocitrate dehydrogenase) and storage (vacuolar proton translocating pumps: one vacuolar H+-ATPase, and two vacuolar H+ pyrophosphatases) were cloned. Five of these peach genes were homologous to genes isolated from fruit in other fleshy fruit species. Phylogenetic and expression analyses suggested the existence of a particular vacuolar pyrophosphatase highly expressed in fruit. The sixth gene was the first cytosolic NAD-dependent malate dehydrogenase gene isolated from fruit. Gene expression was studied during the fruit development of two peach cultivars, a normal-acid (Fantasia) and a low-acid (Jalousia) cultivar. The overall expression patterns of the organic acid-related genes appeared strikingly similar for the two cultivars. The genes involved in organic acid metabolism showed a stronger expression in ripening fruit than during the earlier phases of development, but their expression patterns were not necessarily correlated with the changes in organic acid contents. The tonoplast proton pumps showed a biphasic expression pattern more consistent with the patterns of organic acid accumulation, and the tonoplast pyrophosphatases were more highly expressed in the fruit of the low-acid cultivar during the second rapid growth phase of the fruit. PMID- 11903974 TI - Regulation of the expression of a putative ethylene receptor, PeERS2, during the development of passion fruit (Passiflora edulis). AB - We isolated a full-length cDNA (PeERS2) that encoded the homologue in passion fruit of ERS1 of Arabidopsis and examined its expression during development of passion fruit. PeERS2 was 2357 bp long and included a single open reading frame that encoded a putative protein of 634 amino acids with a calculated molecular mass of 70.8 kDa. Expression of PeERS2 mRNA in arils of passion fruit was enhanced during ripening and after treatment with ethylene, but its level remained very low in seeds over the course of ripening. Accumulation of PeERS2 mRNA in arils was markedly reduced in fruits treated with 2,5-norbornadiene (NBD), but simultaneous application of ethylene abolished the inhibitory effects of NBD, suggesting that the continuous action of ethylene might promote ripening, with a concomitant increase in the abundance of PeERS2 mRNA. Levels of transcripts of the PeERS1 and PeERS2, which encode similar but not identical receptors for ethylene, increased during senescence of flowers and expression of PeERS2 mRNA was also enhanced during formation of the separation layer. The levels of transcripts of PeETR1 (the gene for yet another ethylene receptor) and PeERS1 were, respectively, higher than those of PeERS2 in sepals and ovaries. The transcripts of all three genes for ethylene receptors were barely detectable in anthers. These results suggest that the expression of the three genes for ethylene receptors is differentially regulated and that expression of the gene for PeERS2 is regulated not only by ethylene itself but also by developmental factors. Expression of each of the three individual genes for ethylene receptors might be controlled by different molecular mechanisms in the various tissues. PMID- 11903975 TI - Effects of putrescine accumulation in tobacco transgenic plants with different expression levels of oat arginine decarboxylase. AB - Arginine decarboxylase (ADC; EC 4.1.1.19) is a key enzyme in one of the two possible ways to synthesize putrescine (Put) in plants. In previous work (Masgrau et al. 1997), we observed an altered phenotype (growth inhibition, leaf chlorosis and necrosis) in tobacco transgenic plants (Nicotiana tabacum L. var. Wisconsin 38) containing the oat ADC cDNA under the control of a tetracycline inducible promoter, the severity of which was correlated with Put content. Now we have analysed the T2 generation of a selected transgenic line (line 52), which in previous generations was characterized by presenting a moderate increase in ADC activity and polyamine levels, but no phenotype alterations. Studying two selected individuals, one with a high expression level of the transgene and the other with a moderate expression level, we demonstrate that only the one with increased polyamine content displays the altered (toxic) phenotype. The possible causes of toxicity have been analysed. The results suggest that either Put or its oxidation products, via diamine oxidase (DAO; EC 1.4.3.6), are the responsible factors for the deleterious effects observed in the transgenic plants. PMID- 11903976 TI - Catalase gene expression in response to auxin-mediated developmental signals. AB - The effect of auxin on maize catalase gene expression was examined at several different developmental stages during embryo and seedling development. All three catalase genes and their respective proteins were induced by both natural and synthetic auxin in immature embryos. Total catalase (CAT) activity increased dramatically in response to high concentrations of auxin, with CAT-2, which is not normally expressed at this stage, being the isozyme most responsible for the increase. Cat1 transcript accumulated to high levels at 2-8 h after auxin treatment, while Cat2 and Cat3 transcripts increased dramatically, but only after 12 h. In CAT-2 null mutant lines, the CAT-1 isozyme compensated for the missing CAT-2 activity and was the major isozyme responsible for the observed increase in total CAT activity. Auxin treatment mimics the germination process (i.e. induces germination) in immature embryos. Thus, the observed early induction of CAT-1 and the later increase of CAT-2 during the germination process may be due, in part, to changes in auxin content. In germinating embryos, auxin also induces total CAT activity and Cat transcript accumulation, although to a lesser extent. Auxin also induces Cat1 transcript accumulation in young leaves. The involvement of ROS in the auxin response is discussed. PMID- 11903977 TI - Lignification and lignin heterogeneity for various age classes of bamboo (Phyllostachys pubescens) stems. AB - The lignification process and lignin heterogeneity of fibre, vessel and parenchyma cell walls for various age classes of bamboo stems of Phyllostachys pubescens Mazel were investigated. It was shown that protoxylem vessels lignified in the early stage of vascular bundle differentiation, metaxylem vessel and fibre walls initiated lignification from the middle lamella and cell corners after the completion of vascular bundle differentiation. Most of the parenchyma cell walls lignified after the stem reached its full height, while a few parenchyma cells remained non-lignified even in the mature culm. The cell walls of fibres and most parenchyma cells thickened further during the stem growth to form polylamellate structure and the lignification process of these cells may last even up to 7 years. The fibre walls were rich in guaiacyl lignin in the early stage of lignification, and lignin rich in syringyl units were deposited in the later stage. Vessel walls mainly contained guaiacyl lignin, while both guaiacyl and syringyl lignin were present in the fibre and parenchyma cell walls. PMID- 11903978 TI - Origin and basipetal transport of the IAA responsible for rooting of carnation cuttings. AB - The origin and transport of the IAA responsible for rooting was studied in carnation (Dianthus caryophyllus L.) cuttings obtained from secondary shoots of the mother plants. The presence of mature leaves in the cuttings was essential for rooting. Removal of the apex and/or the youngest leaves did not reduce the rooting percentage as long as mature leaves remained attached. Removal of mature leaves inhibited rooting for a 24-day period during which the basal leaves grew and reached maturity. After this period rooting progressed as in intact cuttings. Auxin (NAA + IBA) applied to the stem base of defoliated cuttings was about 60% as effective as mature leaves in stimulating rooting. Application of NPA to the basal internode resulted in full inhibition of rooting. The view, deduced from these results, that auxin from mature leaves is the main factor controlling the rooting process was reinforced by the fact that mature leaves contained IAA and exported labelled IAA to the stem. The distribution of radioactivity after application of (5-3H)-IAA to mature leaves showed that auxin movement in the stem was basipetal and sensitive to NPA inhibition. The features of this transport were studied by applying 3H-IAA to the apical cut surface of stem sections excised from cuttings. The intensity of the transport was lower in the oldest node than in the basal internode, probably due to the presence of vascular traces of leaves. Irrespective of the localization of the sections and the carnation cultivar used, basipetal IAA transport was severely reduced when the temperature was lowered from 25 to 4 degrees C. The polar nature of the IAA transport in the sections was confirmed by the inhibition produced by NPA. Local application of IAA to different tissues of the sections revealed that polar auxin transport was associated with the vascular cylinder, the transport in the pith and cortex being low and apolar. The present results strongly support the conclusion that IAA originating from the leaves and transported in the stem through the polar auxin transport pathway was decisive in controlling adventitious rooting. PMID- 11903979 TI - Breakage of Pseudotsuga menziesii seed dormancy by cold treatment as related to changes in seed ABA sensitivity and ABA levels. AB - The main aims of the present work were to investigate whether a chilling treatment which breaks dormancy of Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) seeds induces changes in the sensitivity of seeds to exogenous ABA or in ABA levels in the embryo and the megagametophyte, and whether these changes are related to the breaking of dormancy. Dormant seeds germinated very slowly within a narrow range of temperatures (20-30 degrees C), the thermal optimum being approximately 25 degrees C. The seeds were also very sensitive to oxygen deprivation. Treatment of dormant seeds at 5 degrees C improved further germination, and resulted in a widening of the temperature range within which germination occurred and in better germination in low oxygen concentrations. In dry dormant seeds the embryo contained about one-third of the ABA in the megagametophyte. ABA content of both organs increased during the first 4 weeks of chilling. It then decreased sharply in the megagametophyte to the level in the embryo after 7-15 weeks of chilling. At 15 degrees C, a temperature at which dormancy was expressed, the ABA level increased in the embryo and the megagametophyte of dormant unchilled seeds whereas it decreased in the organs of chilled seeds. The longer the chilling treatment, the faster the decrease in ABA after the transfer of seeds from 5 degrees C to higher temperatures, and the decrease was faster at 25 than at 15 degrees C. These results suggest that the breaking of dormancy by cold was associated with a lower capacity of ABA biosynthesis and/or a higher ABA catabolism in the seeds subsequently placed at 15 or 25 degrees C. Moreover, the chilling treatment resulted in a progressive decrease in the sensitivity of seeds to exogenous ABA. However, seeds remained more sensitive to ABA at 15 than at 25 degrees C. The possible involvement of ABA synthesis and of responsiveness of seeds to ABA in the breaking of dormancy by cold treatment is discussed. PMID- 11903980 TI - Carbon dioxide and ethylene interactions in tulip bulbs. AB - The effect of CO2 on ethylene-induced gummosis (secretion of polysaccharides), weight loss and respiration in tulip bulbs (Tulipa gesneriana L.) was investigated. A pretreatment with 1-MCP prevented these ethylene-induced effects, indicating that ethylene action must have been directed via the ethylene receptor. Treatment with 0.3 Pa ethylene for 2 days caused gummosis on 50% of the total number of bulbs of cultivar Apeldoorn, known to be sensitive for gummosis. Addition of CO2 (10 kPa) reduced the ethylene-induced gummosis to 18%. In a second experiment the influence of ethylene and CO2 on respiration and FW loss of bulbs of the cultivar Leen van der Mark was studied. A range of ethylene partial pressures (0.003-0.3 Pa) was applied continuously for 29 days. Ethylene caused a transient peak in O2 consumption rate during the first days after the start of application. The relation between O2 consumption rate and ethylene partial pressure could be described by Michaelis-Menten kinetics. Respiratory peaks were reduced by CO2. This inhibition by CO2 could not totally be due to competition with ethylene at the receptor binding-site, as was indicated by the use of an O2 consumption model. Pre-treatment of bulbs with 1-MCP and subsequent exposure to CO2 showed that CO2 could influence respiration irrespective of any interaction with ethylene. Ethylene and CO2 both stimulated weight loss. The effect of combined treatments of ethylene and CO2 on weight loss was at least as strong as the sum of the separate effects, which implies that competition between ethylene and CO2 at the receptor binding-site was unlikely. PMID- 11903981 TI - The public-private partnership for the Central American handwashing initiative: reflections from a private sector perspective. PMID- 11903983 TI - Improving private practitioner sick-child case management in two urban communities in Pakistan. AB - OBJECTIVE: To evaluate if INFECTOM, a multicomponent behaviour change strategy, would alter the care received by children visiting private healthcare providers so that it was more consistent with the IMCI algorithm. METHODS: Community surveys in two low income communities in Pakistan identified children who had visited healthcare providers in the preceding 2 weeks complaining of diarrhoea, cough or rapid breathing, or fever. Interviewers asked the mothers of these children whether providers performed specific behaviours recommended by the Integrated Management of Childhood Illness (IMCI) algorithm. These data were analysed to generate provider-specific IMCI-related behaviour profiles. A team including community representatives met with the providers, discussed the correct IMCI algorithm behaviour, reviewed the percentage of time each of their practices was consistent with IMCI recommendations, and negotiated a contract with a numerical target for improved practices. This cycle of survey, discussion of results and contracting was repeated three times over 10 months. RESULTS: Twenty two providers, 13 of whom (59%) had a medical degree, regularly treated children in the two communities. Sixteen of the 21 targeted behaviours (76%) occurred with significantly increased frequency during the course of the intervention. Of the 10 practices that ill children with any of the syndromes should have received, at baseline children averaged receiving 4.3. In the final model, each subsequent round of evaluation was associated with a 0.57 increase in the number of appropriate practices performed at visits to non-Bachelor's Degree in Medicine (MBBS) qualified providers (P < 0.001) and a 0.75 increase among visits to MBBS qualified providers (P=0.004). The percentage of children who received an injection decreased from 70 to 56% (P=0.03). CONCLUSIONS: INFECTOM altered the practices of private providers so that they were more consistent with the IMCI algorithm. Efforts to further develop this approach could improve the quality of clinical healthcare in other settings. PMID- 11903982 TI - Case management of malaria in under-fives at primary health care facilities in a Tanzanian district. AB - OBJECTIVE: To study case management of malaria in children under 5 years of age at primary health care facilities in Kibaha district, Tanzania and to evaluate the accuracy of self-reported mothers'/guardians' information on chloroquine use in children. METHOD: A random sample of 652 mothers/guardians with sick children under 5 years of age attending 10 primary health care facilities was observed and interviewed. Blood samples for determination of chloroquine levels were taken from all children and thick smears for detection of malaria parasites were taken from the children who were prescribed chloroquine. Information on diagnoses and prescriptions was collected from recording books. RESULTS: Fever and respiratory problems were the most common complaints and accounted for 75% and 46% of the presenting conditions, respectively (some complained of both). Fifty-four per cent of the children received medication at home, most commonly antipyretics and chloroquine, 20% had been taken to another health facility and 3% to traditional healers before coming to the health facilities. There was a significantly higher use of antipyretics among home treated children compared with those taken previously to health facilities (P or= 1000 nmol/l). Of those prescribed chloroquine, 16% already had high blood concentrations of the drug. CONCLUSION: Health care services, i.e. presumptive malaria diagnosis, consultation time and procedure for physical examination need to be improved. PMID- 11903984 TI - Insecticide-treated bednet use, anaemia, and malaria parasitaemia in Blantyre District, Malawi. AB - OBJECTIVE: To evaluate the use of insecticide-treated bednets and the effectiveness of social marketing for their distribution. METHODS: Systematic cluster sample survey of 1080 households in 36 census enumeration areas across Blantyre district, Malawi, in February 2000. RESULTS: A total of 672 households had one or more children under 5. Bednet ownership was low (20.5% of households) overall, and significantly lower in rural areas than urban areas (6.4 vs. 29.8%, P=0.001). Only 3.3% of rural children under 5 had slept under a net the previous night, compared with 24.0% of urban children (P < 0.001). When asked why they did not own a net, nearly all (94.9%) caretakers in households without nets stated they had no money to buy them. In multivariate statistical models that controlled for the influence of house structure, urban vs. rural location, gender of the head of household, and the primary caretaker's education, rural children under 5 in households without nets experienced a statistically significant higher prevalence of malaria parasitaemia [RR (risk ratio) 4.9, 95% CI (confidence interval) 2.3-10.5] than children in households with at least one bednet. This was also true for urban children under 5 (RR 2.1, 95% CI 1.0-4.2, P=0.04). CONCLUSION: Social marketing approaches to promoting insecticide-treated nets in Blantyre District may have produced measurable health benefits for children in those households in which residents bought and used the products. Market-based approaches may take years to achieve high levels of coverage and may exaggerate inequities between urban and rural populations. PMID- 11903985 TI - Wide distribution of Plasmodium ovale in Myanmar. AB - The presence of Plasmodium ovale has never been previously reported in Myanmar. Using blood samples obtained in many villages across the country between 1996 and 2000, molecular diagnosis of Plasmodium species was made with semi- or full nested polymerase chain reaction (PCR) with species-specific primers, followed by agarose gel electrophoresis to detect amplification products. The presence of P. ovale was also confirmed with the another PCR-based diagnosis, the microtiterplate hybridization (MPH) method using species-specific probes. Both methods target the A type of the small subunit ribosomal RNA gene of the four human malaria parasites. Plasmodium ovale DNA was amplified in samples from 65 (4.9%) of 1323 PCR-positive patients, with perfect agreement between results obtained by nested PCR and MPH. Only four P. ovale-infected patients had single species infection; all others were coinfected with P. falciparum, P. vivax and/or P. malariae. Quadruple infections were observed in six subjects. Parasites with typical P. ovale morphology were found in only 19 patients by conventional microscopy of Giemsa-stained thin smears or fluorescence microscopy of acridine orange-stained thin smears. Plasmodium ovale infections were found in villages situated in the southern, central and western regions of Myanmar, suggesting that P. ovale may be widely distributed in this country. PMID- 11903986 TI - Community factors associated with malaria prevention by mosquito nets: an exploratory study in rural Burkina Faso. AB - Malaria-related knowledge, attitudes and practices (KAP) were examined in a rural and partly urban multiethnic population of Kossi province in north-western Burkina Faso prior to the establishment of a local insecticide-treated bednet (ITN) programme. Various individual and group interviews were conducted, and a structured questionnaire was administered to a random sample of 210 heads of households in selected villages and the provincial capital of Nouna. Soumaya, the local illness concept closest to the biomedical term malaria, covers a broad range of recognized signs and symptoms. Aetiologically, soumaya is associated with mosquito bites but also with a number of other perceived causes. The disease entity is perceived as a major burden to the community and is usually treated by both traditional and western methods. Malaria preventive practices are restricted to limited chloroquine prophylaxis in pregnant women. Protective measures against mosquitoes are, however, widespread through the use of mosquito nets, mosquito coils, insecticide sprays and traditional repellents. Mosquito nets are mainly used during the rainy season and most of the existing nets are used by adults, particularly heads of households. Mosquito nets treated with insecticide (ITN) are known to the population through various information channels. People are willing to treat existing nets and to buy ITNs, but only if such services would be offered at reduced prices and in closer proximity to the households. These findings have practical implications for the design of ITN programmes in rural areas of sub-Saharan Africa (SSA). PMID- 11903987 TI - Level and dynamics of malaria transmission and morbidity in an equatorial area of South Cameroon. AB - We conducted parasitological and entomological malaria surveys among the population of Mengang district in southern Cameroon to analyse the relationship between malaria transmission intensity and malaria morbidity. We investigated two adjacent areas which differ 10-fold in transmission intensity [annual entomological inoculation rate (EIR) 17 vs. 170], but have very similar Plasmodium falciparum malariometric profiles with parasite prevalences of 58 vs. 64%, high parasitaemia prevalences (> 1000 parasites/microl) of 15 vs. 16% and the same morbidity of 0.17-0.5 attacks/person/year. Plasmodium malariae prevalence was 14 vs. 16%. One possible explanation is that the similarity of the duration of the short and high transmission seasons in both areas is equally, if not more, significant for parasitological and clinical profiles as the annual EIR. We discuss the relationships between variations in transmission levels, parasitaemia and clinical incidence, and draw parallels to similar situations elsewhere. PMID- 11903988 TI - Simple method for long-term copro-preservation of Cryptosporidium oocysts for morphometric and molecular analysis. AB - Preservation of Cryptosporidium oocysts in faecal specimens containing 75% ethanol is suitable for subsequent morphometric and molecular analysis. No significant morphologic alteration occurred after storage at ambient temperatures, ranging from 22 to 38 degrees C, for more than 2 years. After washing, sugar floatation and DNA extraction, a nested polymerase chain reaction targeting the small subunit ribosomal RNA gene successfully amplified Cryptosporidium DNA in all 15 isolates examined. The sensitivity of detection by polymerase chain reaction (PCR) was found to be as high as 1.25 oocysts per reaction (mean=3.01, SD=1.14). Importantly, a 2.2-kb of the complete DNA sequence of a gene encoding Cryptosporidium thrombospondin-related adhesive protein (TRAP C1) was also consistently amplified by PCR in all isolates. The PCR-amplified product can be used as a good template for sequencing. Therefore, this simple procedure should be useful for epidemiological analysis of clinical samples from outbreaks, endemic or sporadic cases of cryptosporidiosis when long-term storage of oocysts is required. PMID- 11903989 TI - In vitro activities of ferrochloroquine against 55 Senegalese isolates of Plasmodium falciparum in comparison with those of standard antimalarial drugs. AB - The in vitro activities of ferrochloroquine, chloroquine, quinine, mefloquine, halofantrine, amodiaquine, artesunate, atovaquone, cycloguanil and pyrimethamine were evaluated against Plasmodium falciparum isolates from Senegal (Dielmo, Ndiop), using an isotopic micro-drug susceptibility test. The IC50 values for ferrochloroquine ranged from 0.55 to 28.2 nM and the geometric mean IC50 for the 55 isolates was 7.9 nM (95% CI, 6.5-9.7 nM). Ferrochloroquine was 35 times more active than chloroquine (35-fold greater against chloroquine-resistant isolates), quinine, mefloquine, amodiaquine, cycloguanil and pyrimethamine. Weak positive correlations were observed between the responses to ferrochloroquine and that to chloroquine, quinine, and amodiaquine, but not compulsorily predictive of cross resistance. There was no significant correlation between the response to ferrochloroquine and that to mefloquine, halofantrine, artesunate, atovaquone, cycloguanil and pyrimethamine. Ferrochloroquine may be an important alternative drug for the treatment of chloroquine-resistant malaria. PMID- 11903990 TI - Has directly observed treatment improved outcomes for patients with tuberculosis in southern Thailand? AB - OBJECTIVE: To validate the practice of directly observed treatment (DOT) and evaluate its effect on treatment outcomes. METHODS: This follow-up study conducted in 24 districts in southern Thailand included 411 new, smear-positive, pulmonary tuberculosis (TB) patients who started treatment between February and September 1999. Patients and/or their observers were interviewed about their actual DOT practice during the first 2 months of treatment. Treatment outcomes were evaluated at the end of the second month and at the end of treatment. RESULTS: Of 411 patients, 379 were assigned to DOT but only 68 practised strict DOT for every dose during the first 2 months. Adjusted odds ratios (ORs) for 'no sputum conversion' and 'unsuccessful treatment' were 1.1 (95% CI 0.6-2.1) and 1.3 (95% CI 0.6-2.8), respectively, for those who practised strict DOT vs. the rest. CONCLUSIONS: Actual practice of DOT was quite different from what was intended at the assignment. Practice of strict DOT during the first 2 months was not associated with sputum conversion or treatment success in this study area. PMID- 11903991 TI - Is Rhodnius robustus (Hemiptera: Reduviidae) responsible for Chagas disease transmission in Western Venezuela? AB - We present evidence for the putative role of Rhodnius robustus as extradomestic vector of Chagas disease in Western Venezuela. First, we assessed the validity of this triatomine species by genetic characterization in relation with some other species of the prolixus group. Random amplified polymorphic DNA data showed a clear separation between this species and R. prolixus and indicated a probable genetic heterogeneity within R. robustus. Faeces and gut contents were microscopically examined in 54 of 137 R. robustus collected in palm trees. According to this morphological examination, 18% were positive for Trypanosoma cruzi, 11% harboured T. rangeli and 11% showed mixed infection. Five of the seven samples examined gave a polymerase chain reaction major band of 270 bp specific of T. cruzi. The hybridization probes showed that R. robustus may transmit clones 20 and 39 (or genetically related ones) in Venezuela. Such a transmission might occur when, in absence of domestic R. prolixus and attracted by artificial light, R. robustus enters houses and feeds on humans, or when people are bitten outdoors. The lack of bugs inside houses could mean that the insects leave houses after feeding, or die without reproducing there. PMID- 11903992 TI - Reasons for poor cataract surgery uptake - a qualitative study in rural South Africa. AB - OBJECTIVES: To understand the reasons for poor cataract surgery uptake in people with blindness or severe visual impairment in rural South Africa. METHODS: A qualitative analysis of detailed, domiciliary interviews with a community-based random sample of elderly Zulus who were blind or severely visually impaired as a result of operable cataract, who had previously been invited for surgery but had failed to attend. RESULTS: Fear of surgery and a fatalistic attitude to the inevitability and irreversibility of blindness in old age were the main reasons for failure to attend for surgery. There was a lower level of disability and perceived need than had been assumed for people with such poor visual acuity. Non surgical western style health care for systemic illness was common but few patients had sought any form of assistance for their poor vision. Issues of cost and accessibility were relatively unimportant. CONCLUSION: Provision of affordable and accessible cataract surgery for the blind and severely visually impaired members of a community does not guarantee that it will be taken up. Other barriers to surgery may be revealed when practical issues such as cost and accessibility are addressed. Perceptions of visual disability among subjects with cataract may differ from simple objective clinical standards. PMID- 11903994 TI - Ruth Sanger. PMID- 11903995 TI - Monoclonal anti-D for immunoprophylaxis. AB - Routine antenatal prophylaxis with anti-D has become accepted as desirable, but concerns have been expressed about the adequacies of supply and safety of polyclonal anti-D. Human monoclonal anti-D has been produced using Epstein-Barr virus (EBV)-transformed peripheral B cells, sometimes coupled with fusions to myeloma cell lines. More recently, molecular biology techniques have been used to produce human monoclonal anti-D in a variety of different ways. Many monoclonal antibodies (mAbs) have been characterized for fine specificity and in vitro functional activity in International Workshops. Two mAbs have been shown to cause red cell clearance and immunosuppression in male volunteers. Considerations for the future development of monoclonal anti-D for prophylactic use are reviewed. PMID- 11903996 TI - Early detection of hepatitis B surface antigen and detection of HBsAg mutants: a comparison of five assays. AB - BACKGROUND AND OBJECTIVES: Assays for the detection of hepatitis B surface antigen (HBsAg) face two challenges: the emergence of sensitive techniques for detection of hepatitis B virus DNA and the existence of HBsAg mutant hepatitis B viruses. We studied the sensitivity of five modern assays for detection of HBsAg. MATERIALS AND METHODS: The sensitivity of two mini robot-based assays--IMx HBsAg V2 and Vidas HBsAg--and three enzyme-linked immunosorbent assays (ELISAs) with microtitre plate format--Hepanostika Uni-Form II v1.2, Monolisa Ag HBs Plus and HBsAg Test System 3--was compared testing 11 HBsAg seroconversion series, serial dilutions prepared from three HBsAg standards and nine HBsAg mutant samples. RESULTS AND CONCLUSION: Overall, the IMx HBsAg V2 assay showed the highest sensitivity. It outperformed the Vidas HBsAg in analytical sensitivity and in detection of HBsAg mutants. Among the microtitre plate ELISAs, the Monolisa Ag HBs Plus outperformed the HBsAg Test System 3 in detection of HBsAg mutants and it surpassed the Hepanostika Uni-Form II v1.2 in analytical sensitivity. PMID- 11903997 TI - Hepatitis B core antibodies in Danish blood donors: a surrogate marker of risk behaviour. AB - BACKGROUND AND OBJECTIVES: The aim of this work was to determine the prevalence of antibodies to hepatitis B core antigen (anti-HBc) among Danish blood donors and to correlate this with risk factors for blood-borne and sexually transmitted diseases. MATERIALS AND METHODS: During a 5-month period, 10 862 consecutive donors in the County of Funen were screened for anti-HBc, and repeat-reactive samples were confirmed by supplementary testing. Information on risk factors was assessed by questionnaire in 585 consecutive anti-HBc-negative blood donors and compared with information obtained from confirmed positive donors. RESULTS: The prevalence of confirmed positive anti-HBc among donors was 0.70% (76/10 862, 95% confidence interval [CI]: 0.55-0.87). One donor was positive for anti-HBc immunoglobulin M (IgM); none tested positive for hepatitis B virus (HBV) DNA. In a logistic regression analysis, age, female gender, tattoos and commercial sexual relations, were independent predictive factors for the presence of anti-HBc. CONCLUSION: Anti-HBc is a surrogate marker for previous risk behaviour in the Danish blood donor population. We suggest that screening for anti-HBc may be used among new donors to supplement interviews on risk behaviour. PMID- 11903998 TI - Parvovirus B19 DNA in plasma pools and plasma derivatives. AB - BACKGROUND AND OBJECTIVES: Human parvovirus B19 (B19) has been transmitted by various plasma-derived medicinal products. The aim of this study was to determine the frequency and the level of B19 DNA contamination in plasma pools destined for fractionation and in a broad range of plasma derivatives. In addition, removal of B19 DNA by the manufacturing process was investigated in cases where corresponding samples from plasma pool and product were available. MATERIALS AND METHODS: Plasma pool samples and blood products were tested for B19 DNA by nested polymerase chain reaction (PCR), and the viral DNA content was determined by TaqMan quantitative PCR. RESULTS: Two-hundred and twenty two of 372 plasma pools for fractionation contained B19 DNA at concentrations of 10(2)-10(8) genome equivalents/ml (geq/ml). While approximately 65% of the DNA-positive plasma pools were only moderately contaminated (< 10(5) geq/ml), 35% contained > 10(6) geq/ml. High frequencies of contamination were detected in Factor VIII (79 of 91), prothrombin complex concentrates (38 of 43) and Factor IX (41 of 62), where the concentration of B19 DNA ranged between 102 and 107 geq/ml. A lower level of B19 DNA contamination was found in antithrombin III (five of 26 samples), in anti-D immunoglobulins (three of 37 samples) and in albumin (four of 51 samples), with levels ranging between 10(2) and 10(3) geq/ml. Furthermore, investigation of plasma pools for solvent/detergent plasma (S/D plasma), from two manufacturers, revealed B19 DNA in 15 of 66 batches at concentrations of 10(2)-10(8) geq/ml. Similar concentrations were detected in the corresponding final S/D plasma products. Anti-B19 immunoglobulin G (IgG) was found in plasma pools and S/D plasma at concentrations of approximately 40 IU/ml. CONCLUSION: Although positive PCR results do not necessarily reflect infectivity, these data show that B19 is a common contaminant in plasma pools and in plasma-derived medicinal products. Considering the resistance of animal parvoviruses to inactivation by heat and chemical agents, and the absence of specific information for B19, the risk of B19 transmission by plasma products should be considered. Physicians should be aware of this problem when treating patients of B19-related risk groups. The plasma fractionation industry should continue their efforts to avoid B19 contamination of plasma derivatives and develop methods which are effective in removing/inactivating parvovirus B19. PMID- 11903999 TI - Species differences in the blood content of the normal cellular isoform of prion protein, PrP(c), measured by time-resolved fluoroimmunoassay. AB - BACKGROUND AND OBJECTIVES: The concern that variant Creutzfeldt-Jakob disease could be transmitted via blood transfusion has prompted studies of blood infectivity in animal models. As normal prion protein acts as a substrate for conversion to the abnormal form associated with infectivity, we have quantified its distribution in mice and hamsters, the most commonly used animal models. MATERIALS AND METHODS: A time-resolved fluoroimmunoassay was used to measure normal prion protein in hamster and mouse tissues, including blood. RESULTS: Levels of prion protein in hamster blood were remarkably low compared with human blood. In contrast, levels in mouse blood were quite similar to human blood; however, there were differences in the distribution of normal prion between cellular and cell-free fractions. CONCLUSION: Differences between levels of normal prion in blood of animal models and humans should be considered as a possible contributor to infectivity study outcomes in these models. PMID- 11904000 TI - Loss of phosphotyrosine phosphatase activity and changes in the tyrosine phosphorylation state of proteins after storage of sheep platelets in plasma or Seto solution at 4 degrees C. AB - BACKGROUND AND OBJECTIVES: During platelet storage an array of deleterious changes occur, through mechanisms not fully understood, which impair platelet haemostasis. Transfused platelets should maintain the integrated networks of signalling pathways that regulate platelet activation and functionality. We hypothesized that protein phosphorylation and dephosphorylation, which play a fundamental role in these pathways, might be affected by platelet storage. We therefore investigated whether the activity of phosphotyrosine phosphatase (PTP), which belongs to an oxidant-susceptible group of enzymes involved in the platelet signal-transduction pathways that ensure platelet functionality, is affected by platelet storage. MATERIALS AND METHODS: Using sheep platelet species as a model system, we conducted serial studies on the membranes of platelets and microparticles shed during platelet storage, in their own plasma or in a synthetic medium called Seto, for up to 5 days at 4 degrees C. RESULTS: A progressive decrease in both total and specific membrane-associated PTP activities from whole platelets (but not from microparticles) located within each platelet storage bag was observed from day 1 onwards in both types of storage media. These decreases could be partly avoided by the addition of vitamin E. Additionally, the observed decrease in PTP activity was accompanied with increases in the tyrosine phosphorylation of proteins from whole platelets or crude platelet membranes, the tyrosine phosphorylation state of proteins from microparticles remaining basically unchanged. CONCLUSION: Our findings suggest that alterations of at least the tyrosine phosphorylation balance might be one of the reasons for the decrease in the haemostatic function of stored platelets. PMID- 11904001 TI - Effectiveness of a protocol to improve transfusion practice in knee replacement surgery. AB - BACKGROUND AND OBJECTIVES: The available guidelines on red cell transfusion are not clearly defined and therefore have had only a modest impact on transfusion practice. The aims of this study were to assess the rate of compliance with a transfusion algorithm and its effect on transfusion practice. MATERIALS AND METHODS: A prospective observational study was carried out on 101 patients who underwent primary elective total knee replacement surgery. RESULTS: Only 30% of the patients were transfused. An overall compliance rate of 77% was achieved with the transfusion algorithm. CONCLUSION: A transfusion algorithm, together with staff education, is effective in reducing both the number of patients transfused and inappropriate transfusions. PMID- 11904002 TI - Molecular characterization of weak D phenotypes by site-directed mutagenesis and expression of mutant Rh-green fluorescence protein fusions in K562 cells. AB - BACKGROUND AND OBJECTIVES: Mutations detected in 161 weak D samples from Caucasians have been classified into 16 types. Because flow cytometry using monoclonal anti-D antibodies (mAbs) has shown that weak D red cells display type specific antigen density, these mutations in transmembranous regions have been assigned weak D phenotypes. The present study attempts to confirm or refute this assignment. MATERIALS AND METHODS: We amplified DNA from four Japanese weak D samples using the polymerase chain reaction (PCR), and directly sequenced the amplified DNA. Using site-directed mutagenesis, we constructed three vectors expressing mutant RHDs-- G212C, V270G (weak D type 1) and G358A (type 2)--in K562 cells. The expression of RhD antigens was examined by flow cytometry using mAbs. RESULTS: A new mutation resulting in a conversion at amino acid residue 212 (Gly to Cys) was detected in a Japanese weak D sample. K562 cells transduced with mutant RhD cDNA reacted weakly in a type-specific manner with mAbs. CONCLUSIONS: The mutations--G212C (new weak D type), V270G (weak D type 1) and G358A (type 2)- in transmembranous regions had obvious effects on the D epitopes recognized by mAbs. The results of this study provide direct evidence that these mutations can account for weak D phenotypes. PMID- 11904004 TI - GB virus-C/hepatitis G virus and transfusion-transmitted virus infection in blood donors in a tertiary care hospital in south India. PMID- 11904003 TI - Point mutations in KEL exon 8 determine a high-incidence (RAZ) and a low incidence (KEL25, VLAN) antigen of the Kell blood group system. AB - BACKGROUND AND OBJECTIVES: The molecular basis of two Kell blood group antigens, RAZ (provisionally KEL27) and VLAN (KEL25), were determined. MATERIALS AND METHODS: The DNA sequences of the open reading frames and the flanking intron regions of the 19 KEL exons from RAZ and VLAN probands were compared with that of common KEL. Genotyping assays were designed to confirm and detect RAZ and VLAN phenotypes. RESULTS: A homozygous G865A mutation, encoding lysine instead of glutamic acid at amino acid position 249 of Kell protein, defines the RAZ phenotype, while a heterozygous G863A mutation in KEL, encoding an arginine to glutamine substitution at amino acid 248, characterizes the VLAN phenotype. CONCLUSION: Point mutations G865A and G863A, in adjacent codons of KEL exon 8, which cause amino acid substitutions, characterize the RAZ and VLAN Kell blood group phenotypes. PMID- 11904005 TI - Receptor-mediated haemagglutination screening and reduction in the viral load of parvovirus B19 DNA in immunopurified Factor VIII concentrate (Cross Eight M). PMID- 11904006 TI - Transfusion-related acute lung injury (TRALI). PMID- 11904017 TI - Steal affecting the central nervous system. AB - Steal is a pathophysiological process in which increased blood flow through a low resistance vascular bed is sufficient to divert flow away from a region of the central nervous system. Three disease states in which steal may cause neurological deficits due to central nervous system ischemia are reviewed. Subclavian steal occurs when stenosis of the subclavian artery proximal to the vertebral origin causes retrograde flow in the left vertebral artery. Patients with anatomic subclavian steal usually do not develop neurological symptoms but may rarely present with posterior circulation ischemia. Arteriovenous malformations alter cerebral blood flow patterns and regional perfusion pressure. It has been hypothesized that cerebral arteriovenous malformations may cause neurological deficits due to steal and that these deficits may be cured with arteriovenous malformation treatment. Intra-arterial pressure measurements and transcranial velocity studies show regional hemodynamic alterations. However, these changes have not been correlated with presenting symptoms. Evidence from single-photon emission computed tomography does suggest a relationship between regional hypoperfusion and neurological deficits. Coarctation of the aorta may divert flow from the spinal cord circulation through intercostal arteries distal to the stenosis. This is a possible but unproven mechanism of myelopathology. Steal syndromes may be amenable to treatment by open surgical or endovascular approaches. Experimental studies of the pathophysiology of steal are strengthened by precise definitions of the measured parameters and innovative applications of technology. PMID- 11904018 TI - New trends in neuromodulation for the management of neuropathic pain. AB - Since its first application in 1967, the methodology and technology of spinal cord stimulation for the management of chronic, intractable pain have evolved continuously. Despite these developments and improved knowledge of the effects of spinal anatomy and epidural contact configuration on paresthesia coverage, the clinical results of spinal cord stimulation-particularly the long-term effects are still unsatisfactory in many patients. This dissatisfaction has come primarily from the failure of single-electrode configurations to provide consistent paresthesia coverage of the entire painful area. Therefore, new approaches were developed during the late 1990s that attempted to selectively cover one or more dermatomes with paresthesia as well as to provide sequential stimulation of different anatomic sites. These approaches have been applied both intraspinally and extraspinally by stimulating either the spinal nerves or the dorsal columns. To target parts of the latter, different methods have been developed and tested using either two-dimensional contact configurations or electronic field steering. These developments hold promise for improving long term outcomes as well as increasing the number of pain conditions that can be treated with neuromodulation therapy. In this review, the history, theoretical basis, and evolution of these methodologies, as well as the ways in which they represent new trends in neuromodulation, are discussed. PMID- 11904019 TI - Microvascular decompression in the management of glossopharyngeal neuralgia: analysis of 217 cases. AB - OBJECTIVE: Glossopharyngeal neuralgia (GPN) is a rare condition that often presents with the seemingly benign symptom of deep throat pain. Medical management of this condition has not been very effective, and surgical therapy has ranged from nerve sectioning to microvascular decompression (MVD). We present our experience with more than 200 patients who underwent MVD for treatment of GPN at our institution. METHODS: We conducted a retrospective review of our database and identified patients who presented for treatment of presumed GPN. When possible, patients were contacted by telephone for collection of follow-up information regarding symptom relief, complications, functional outcomes, and patient satisfaction. Univariate and multivariate analyses were performed to identify predictors of good outcomes after MVD. Subgroup analyses were performed with quartiles of approximately 50 patients each, for assessment of the effects of improvements in techniques and anesthesia during this 20-year period. RESULTS: We observed GPN to be more common among female (66.8%) than male (33.2%) patients, with an overall mean patient age of 50.2 years (standard deviation, 14.4 yr). The most common presenting symptoms were throat and ear pain and throat pain alone, and the mean duration of symptoms was 5.7 years (standard deviation, 5.8 yr; range, 1-32 yr). Symptoms appeared almost equally on the left side (54.8%) and the right side (45.2%). The overall immediate success rate exceeded 90%, and long-term patient outcomes and satisfaction were best for the typical GPN group (with pain restricted to the throat and palate). Complication rates decreased across quartiles for all categories evaluated. CONCLUSION: MVD is a safe, effective form of therapy for GPN. It may be most beneficial for patients with typical GPN, especially when symptoms are restricted to deep throat pain only. PMID- 11904020 TI - Microvascular decompression to treat hemifacial spasm: long-term results for a consecutive series of 143 patients. AB - OBJECTIVE: The concept of neurovascular decompression for the treatment of hemifacial spasm is now widely accepted. In this study, we report our long-term results for 145 cases treated with this procedure. METHODS: The results of 145 microvascular decompressions to treat hemifacial spasm (performed between 1980 and 1998) among 143 patients (62.2% female patients and 37.8% male patients; mean age, 54.5 yr) are presented. The onset of symptoms was typical in 95.9% of cases and atypical in 4.1%. Platysma muscle involvement was observed for 24.5% of patients, with a higher incidence among female patients (74.3%). Patients were monitored with annual questionnaires. Twenty-six patients were lost to follow-up monitoring, and 117 are still undergoing follow-up monitoring, with an average period of 9.6 years (range, 1-17.6 yr). RESULTS: At discharge, 69 patients (59%) were spasm-free and 48 patients (41%) experienced further spasm. At 6 months, the number of spasm-free patients had increased to 108 (92.3%), whereas only 9 patients (7.7%) complained of hemifacial spasm; 44 patients were spasm-free at an average time of 15 weeks. In follow-up examinations (average period, 9.4 yr), 106 patients were spasm-free. Seven patients experienced only temporary relief, with recurrence after 4.5 years. Two patients were spasm-free after 4 or 6 weeks, and the recurrence of spasm was observed 1 year later. Two patients were never completely spasm-free. Among the patients who did not undergo previous surgery elsewhere, only two experienced recurrence. CONCLUSION: Deafness was the main postoperative complication (8.3%); most of those cases (66%) occurred before the routine use of intraoperative evoked potential monitoring. Analysis of our series demonstrates that this surgical procedure involves very low risk, is well tolerated by elderly patients, is associated with very low recurrence rates, and is a definitive treatment for more than 90% of cases. PMID- 11904021 TI - Role of postoperative magnetic resonance imaging after microvascular decompression of the facial nerve for the treatment of hemifacial spasm. AB - OBJECTIVE: This study was performed to investigate the role of postoperative three-dimensional short-range magnetic resonance angiography in the prediction of clinical outcomes after microvascular decompression (MVD) for the treatment of hemifacial spasm. METHODS: We examined pre- and postoperative magnetic resonance imaging scans obtained between March 1999 and May 2000 for 122 patients with hemifacial spasm, to evaluate the degree of detachment of the vascular contact and changes in the positions of offending vessels. The degree of vascular decompression of the facial nerve root was classified into three groups, i.e., contact, partial decompression, or complete decompression. Contact was defined as unresolved compression, as indicated by postoperative three-dimensional short range magnetic resonance angiography. Partial decompression was defined as incompletely resolved compression; vascular indentation of the facial nerve was improved, but contact with the facial nerve remained. Complete decompression was defined as completely resolved compression. These findings were compared with the surgical findings and clinical outcomes. RESULTS: Of 122 patients with MVD, complete decompression of offending vessels at the root entry zone of the facial nerve was observed for 106 patients (86.9%), partial decompression was observed for 10 patients (8.2%), and contact with offending vessels was observed for 6 patients (4.9%) by using postoperative three-dimensional short-range magnetic resonance angiography. Our study demonstrated that the types of offending vessels affected neither the degree of decompression of the root entry zone of the facial nerve nor surgical outcomes (P > 0.05). Also, there was no significant relationship between the degree of decompression and improvement of symptoms (P > 0.05). Furthermore, there was no significant relationship between the degree of decompression and the timing of symptomatic improvement (P > 0.05). CONCLUSION: Our data suggest that MVD of the facial nerve alone may not be sufficient to resolve symptoms for all patients with hemifacial spasm. Therefore, unknown factors in addition to vascular compression may cause symptoms in certain cases, and it may be necessary to remove those factors, simultaneously with MVD, to obtain symptom resolution. PMID- 11904022 TI - Delayed surgical resection of central nervous system germ cell tumors. AB - OBJECTIVE: To determine the value of delayed surgical resection in patients with central nervous system germ cell tumors who exhibit less than complete radiographic response despite declining serum and cerebrospinal fluid (CSF) tumor markers after initial chemotherapy. METHODS: We retrospectively analyzed 126 patients enrolled on two international multicenter clinical trials (the First and Second International Central Nervous System Germ Cell Tumor Studies) for patients with newly diagnosed central nervous system germ cell tumors. After at least three cycles of chemotherapy, 10 of these patients underwent delayed surgical resection owing to evidence of residual radiographic abnormalities despite declining or completely normalized serum and CSF levels of alpha-fetoprotein and human chorionic gonadotropin. RESULTS: Eight of these patients demonstrated nongerminomatous germ cell tumor elements at the time of initial diagnosis. In these patients, either serum or CSF tumor markers were elevated initially. Two patients demonstrated pure germinomas with normal levels of serum and CSF tumor markers. After chemotherapy, radiographic evaluation revealed a partial response in seven patients, a minor response in one patient, and stable disease in two patients. All 10 patients had either normal or decreasing levels of serum and CSF tumor markers before second-look surgery. At delayed surgical resection, 7 of the 10 patients underwent gross total resection, and 3 patients underwent subtotal resection of residual lesions. Pathological findings at second-look surgery demonstrated three patients to have mature teratomas, two with immature teratomas, and five with necrotic or scar tissue alone. To date, 7 of the 10 patients have had no recurrence during an average follow-up time of 36.9 months (range, 3-96 mo). Three of four patients with nongerminomatous germ cell tumors who had tumor markers that were decreased, but not normalized, before second-look surgery eventually developed tumor dissemination/progression, and they required subsequent radiation therapy despite having teratoma or necrosis/scar tissue at delayed surgery. In contrast, three of four patients with nongerminomatous germ cell tumors and completely normalized markers did not progress and did not require radiation therapy. CONCLUSION: Delayed surgical resection should be considered in patients with central nervous system germ cell tumors who have residual radiographic abnormalities and normalized tumor markers, because these lesions are likely to be teratoma or necrosis/scar tissue. However, second-look surgery should be avoided in patients whose tumor markers have not normalized completely. PMID- 11904023 TI - Management strategy for adult patients with dorsal midbrain gliomas. AB - OBJECTIVE: Dorsal midbrain gliomas (DMGs) involving the tectum occur more commonly in children than in adults. These lesions are often indolent in nature, and patients require treatment only for obstructive hydrocephalus. Because limited information is available concerning adults with this type of lesion, we describe our experience and management strategy in adults with DMGs. METHODS: We retrospectively reviewed the charts and magnetic resonance imaging scans of five adult patients (four men, one woman; mean age, 51.6 yr; range, 23-69 yr) who were treated from March 1992 to August 2001 for DMGs involving the tectum. The mean follow-up time for these patients was 71.4 months (range, 25-113 mo). We analyzed the data to determine the optimal treatment strategy and outcomes of patients with DMGs. Tumor volume was analyzed objectively with Scion Image software (Scion Corp., Frederick, MD) to document changes in volume and determine whether treatment strategy differed significantly with tumor size. RESULTS: Tumors in two of these patients were found incidentally, and three others presented with obstructive hydrocephalus. Magnetic resonance imaging scans demonstrated an isolated tectal glioma in one patient, tectal and tegmental (periaqueductal) gliomas in three patients, and a tectal glioma with right thalamic extension in one patient. Treatment consisted of routine follow-up for the two asymptomatic patients and cerebrospinal fluid diversion surgery for the three patients with hydrocephalus. Volumetric analysis demonstrated that all asymptomatic patients had tumors smaller than 9.3 cm(3), and symptomatic patients had tumors larger than 28.5 cm(3). All follow-up magnetic resonance imaging scans revealed stable dorsal midbrain lesions, and no patient required tumor-specific therapy. CONCLUSION: Although tumors of the dorsal midbrain occur primarily in the pediatric population, similar tumors may occur in adults. As has been learned from experience with children, these lesions are often clinically and radiographically stable and require only appropriate therapy for obstructive hydrocephalus. We advocate a similar conservative strategy of routine imaging follow-up and treatment for hydrocephalus in adult patients. Surgery and other therapy should be reserved for patients with progressive lesions. PMID- 11904024 TI - Diffusion-weighted magnetic resonance imaging in patients with subarachnoid hemorrhage. AB - OBJECTIVE: To evaluate the occurrence and distribution of direct brain injury caused by acute subarachnoid hemorrhage (SAH) by the use of magnetic resonance imaging. METHODS: Computed tomography and magnetic resonance imaging, including diffusion-weighted imaging (DWI), were performed in 32 patients with SAH by use of a 1.5-T whole-body superconductive scanner equipped with an echo planar imaging system. In all cases, computed tomographic and magnetic resonance imaging scans were obtained at the time of admission, before angiography and surgical intervention. RESULTS: No abnormalities were revealed by DWI in any of the low grade SAH patients. However, five (71%) of seven patients diagnosed as having poor-grade SAH (World Federation of Neurosurgical Societies Grades 4 and 5) displayed multiple, patchy focal abnormalities on DWI. Computed tomographic scans obtained at admission failed to clearly demonstrate all of the damaged areas of the brain that were visualized by DWI. These lesions were located in supratentorial cerebral parenchyma, but not in the thalamus, basal ganglia, or cerebellar hemisphere. These multiple widespread lesions exhibiting laminar involvement of the cerebral cortex were not associated with the site of the ruptured aneurysm. CONCLUSION: DWI revealed widespread multifocal lesions in the cerebral cortex of acute poor-grade SAH patients. DWI provides accurate images of all areas of brain damage directly attributable to SAH. PMID- 11904025 TI - Prognostic significance of hypernatremia and hyponatremia among patients with aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: Abnormal serum sodium levels (hyponatremia and hypernatremia) are frequently observed during the acute period after aneurysmal subarachnoid hemorrhage (SAH) and may worsen cerebral edema and mass effect. We performed this study to determine the prognostic significance of serum sodium concentration abnormalities. METHODS: We analyzed prospectively collected data for the placebo treatment group in a clinical trial conducted at 54 neurosurgical centers in North America. The presence of hypernatremia (serum sodium concentration of >145 mmol/L) and hyponatremia (serum sodium concentration of <135 mmol/L) was determined with serum sodium measurements obtained at admission and 3, 6, and 9 days after SAH. The effects of hypernatremia and hyponatremia on the risk of symptomatic vasospasm and on 3-month outcomes were analyzed after adjustment for the following potential confounding factors: age, sex, preexisting hypertension, admission Glasgow Coma Scale score, initial mean arterial pressure, subarachnoid clot thickness, intraventricular blood or intraparenchymal hematoma, ventricular dilation, and aneurysm size and location. RESULTS: Of 298 patients in the analysis, 58 (19%) developed hypernatremia and 88 (30%) developed hyponatremia. Hypernatremia was significantly associated with poor outcomes (odds ratio, 2.7; 95% confidence interval, 1.2-6.1). A positive correlation was observed between the highest sodium values recorded and Glasgow Outcome Scale scores at 3 months (P < 0.0001 by analysis of variance). Hyponatremia was not associated with 3 month outcomes (odds ratio, 1.9; 95% confidence interval, 0.9-4.3). Neither hypernatremia nor hyponatremia was associated with the risk of symptomatic vasospasm. CONCLUSION: Hyponatremia seems to be more common than hypernatremia after SAH. However, hypernatremia after SAH is independently associated with poor outcomes, and this association is independent of previously identified outcome predictors, including age and admission Glasgow Coma Scale scores. Further studies are needed to define the underlying mechanism of this association. PMID- 11904027 TI - There is no transmantle pressure gradient in communicating or noncommunicating hydrocephalus. AB - OBJECTIVE: To examine whether a transmantle pressure gradient exists in adult patients with communicating and noncommunicating hydrocephalus. METHODS: Ten patients participated in the study. The mean patient age was 57 +/- 18 years (range, 20-80 yr); seven patients had communicating hydrocephalus, and three had noncommunicating hydrocephalus. Microsensors were used to measure the intracranial pressure (ICP), for 17 to 24 hours during sleeping and waking periods, in the right lateral ventricle (ICP(IV)) and in the subarachnoid space (ICP(SAS)) over the right cerebral convexity simultaneously. Patient activities and body positions were documented. The hydrostatic pressure difference between the two sensors was calculated from cranial x-rays for four basic body positions and compared with the actual body positions of the patients and the measured difference between the two sensors. For three 10-minute periods, the exact transmantle pressure gradient was calculated for each patient as ICP(IV) - ICP(SAS), adjusted for the hydrostatic pressure difference. RESULTS: The measured pressure difference between the two sensors was always within the limits of the maximal possible hydrostatic pressure difference, and it correlated well with the expected difference for the various body positions: mean correlation coefficient, 0.79 +/- 0.10 (range, 0.65-0.92). The exact mean transmantle pressure was -0.01 +/- 0.24 mmHg (range, -0.4 to 0.4 mmHg). ICP waves caused by cardiac pulse, respiration, and B waves were identical in both spaces. CONCLUSION: This study demonstrates no factual support for existence of a transmantle pressure gradient in nonacute communicating or noncommunicating hydrocephalus. PMID- 11904026 TI - Characteristics of ventricular shunt malfunctions among patients with neurocysticercosis. AB - OBJECTIVE: Ventricular shunts used to relieve hydrocephalus among patients with neurocysticercosis have been plagued by high shunt malfunction rates. We examined the characteristics of shunt malfunctions among patients with neurocysticercosis. METHODS: This is a retrospective chart review of data for 122 patients who were admitted with a diagnosis of cysticercosis during a 5.5-year period. Cases of hydrocephalus requiring shunt placement were reviewed with respect to the segment of shunt obstruction, disease activity at the time of shunt placement, and the effects of antihelminthic treatment on shunt failures and longevity. RESULTS: Twenty-one patients required 49 operations for relief of hydrocephalus, including 22 new shunt placements, 23 revisions, and 4 cyst extirpations. Of these shunt failures, 78% occurred within the first 12 months and 96% within 3 years. Of the failures that occurred in the distal segment, 75% occurred within 6 months. By comparison, 33% of proximal segment obstructions and 50% of the total number of valve obstructions occurred within the first 6 months. Of the shunts placed during the vesicular stage of infection, 63% required revisions, compared with 29% of those placed during the colloidal through calcified stages. Nineteen shunts were placed during the vesicular stage, and nine patients received a full course of antihelminthic treatment after shunt placement. In less than 6 months, 33% of the cases involving shunt placement followed by antihelminthic treatment exhibited shunt failure, compared with 90% of the cases without antihelminthic treatment (P < 0.05, chi(2) test). CONCLUSION: Among patients with vesicular stage cysticercosis, placement of a ventriculoperitoneal shunt followed by a course of antihelminthic medication seems to promote shunt longevity. PMID- 11904029 TI - Blood flow and ischemia within traumatic cerebral contusions. AB - OBJECTIVE: To provide evidence of irreversible ischemia in cerebral contusions among patients with severe traumatic brain injuries and to clarify the potential viability of tissue in the pericontusional zone, quantitative regional cerebral blood flow (rCBF) measurements obtained with the xenon-enhanced computed tomographic method were correlated with the areas of contusions, by using image fusion. METHODS: rCBF measurements obtained during the acute phase (mean, 2 d after injury; range, 0-10 d) were statistically correlated with the extent of tissue necrosis identified as focal atrophy on late follow-up computed tomographic scans (mean time after the xenon-enhanced computed tomographic cerebral blood flow investigation, 265 d; range, 30-1047 d). RESULTS: Seventeen patients exhibited 26 traumatic contusions. All contusions progressed to late focal atrophic areas on the follow-up computed tomographic scans. The rCBF values within the traumatic contusions ranged from 0.5 to 22.0 ml/100 g/min, with a mean of 5.9 +/- 5.9 ml/100 g/min. The contusions exhibited a specific rCBF profile, presenting as a core of severe lethal ischemia surrounded by variable but gradually increasing perfusion with increasing distance from the ischemic core. CONCLUSION: The ischemic profile of the contusions, with a pericontusional zone of low rCBF, presents the potential risk of secondary ischemic insults, similar to the risk in the ischemic penumbral zones surrounding areas of acute ischemic stroke. PMID- 11904028 TI - Cerebral extraction of oxygen and intracranial hypertension in severe, acute, pediatric brain trauma: preliminary novel management strategies. AB - OBJECTIVE: To evaluate long-term clinical outcomes after severe, acute, pediatric brain trauma, in relation to cerebral extraction of oxygen (CEO(2)) and intracranial pressure abnormalities treated with a protocol to simultaneously normalize both parameters. METHODS: Forty-five acutely comatose children who had sustained severe, non-missile brain trauma were prospectively evaluated and treated according to a protocol to maintain normalized values not only for intracranial pressure and perfusion pressure but also for CEO(2) (the arteriojugular oxyhemoglobin saturation difference). Six-month clinical outcomes were assessed in relation to physiological abnormalities observed during the acute phase of injury. RESULTS: At 6 months after injury, 37 children (82.2%) had achieved favorable clinical outcomes, whereas eight children (17.8%) had not. The mortality rate was 4.4% (two children only). For the overall series, intracranial hypertension was closely associated with the development of relative cerebral hyperperfusion (decreased CEO(2)), especially after postinjury Day 1. A comparison of data for children with favorable versus unfavorable clinical outcomes revealed statistically significant between-group differences for high intracranial pressure and low CEO(2) values, both of which were more prominent in the unfavorable outcome group. No significant within- or between-group differences with respect to blood pressure were observed. CONCLUSION: In severe, acute, non-missile pediatric brain trauma, phasic physiological patterns demonstrated an association between the development of intracranial hypertension and relative cerebral hyperperfusion (decreased global CEO(2)), especially after postinjury Day 1. Unfavorable clinical outcomes were significantly related to more pronounced intracranial hypertension and more profound concomitant decreases in CEO(2), indicating hyperoxic uncoupling between global cerebral consumption of oxygen and cerebral blood flow. PMID- 11904030 TI - Quantitative assessment of surgical decompression of the cervical spine with cine phase contrast magnetic resonance imaging. AB - OBJECTIVE: We measured cerebrospinal fluid flow velocity by use of cine phase contrast magnetic resonance imaging to quantitate the effectiveness of surgical decompression in patients with cervical myelopathy. METHODS: Forty-seven patients with cervical myelopathy attributable to either spondylosis or ossification of the posterior longitudinal ligament were studied. Thirty-five patients underwent anterior cervical decompression and fusion; 12 others underwent expansive laminoplasty. Patients were examined preoperatively and postoperatively by use of a 1.5-T scanner with a pulse-gated cine phase contrast sequence. Cerebrospinal fluid flow direction and velocity in the ventral subarachnoid space were determined at the C1 and T1 levels. Forty-four healthy control subjects were examined to determine normal flow velocity parameters. Severity of cervical myelopathy was evaluated preoperatively and postoperatively by use of Japan Orthopedic Association scores to calculate the extent of recovery. RESULTS: Preoperatively, cerebrospinal fluid flow velocity in the caudal direction was significantly lower at both C1 and T1 than velocities measured in healthy controls. Both decompressive procedures essentially returned patient velocities to control values. Clinical recovery from myelopathy did not differ between anterior and posterior decompression. Postoperative increase in flow velocity correlated with clinical recovery after posterior (P < 0.0008) but not anterior decompression. CONCLUSION: Cine phase contrast magnetic resonance imaging provides quantitative assessment of cervical spine decompression, with particularly good clinical applicability to posterior procedures. PMID- 11904031 TI - Laser surface scanning for patient registration in intracranial image-guided surgery. AB - OBJECTIVE: To report our clinical experience with a new laser scanning-based technique of surface registration. We performed a prospective study to measure the calculated registration error and the application accuracy of laser surface registration for intracranial image-guided surgery in the clinical setting. METHODS: Thirty-four consecutive patients with different intracranial diseases were scheduled for intracranial image-guided surgery by use of a passive infrared surgical navigation system. Surface registration was performed by use of a Class I laser device that emits a visible laser beam. The Polaris camera system (Northern Digital, Waterloo, ON, Canada) detects the skin reflections of the laser, which the software uses to generate a virtual three-dimensional matrix of the anatomy of each patient. An advanced surface-matching algorithm then matches this virtual three-dimensional matrix to the three-dimensional magnetic resonance therapy data set. Registration error as calculated by the computer was noted. Application accuracy was assessed by use of the localization error for three distant anatomic landmarks. RESULTS: Laser surface registration was successful in all patients. For the surgical field, application accuracy was 2.4 +/- 1.7 mm (range, 1-9 mm). Application accuracy was higher for the surgical field of frontally located lesions (mean, 1.8 +/- 0.8 mm; n = 13) as compared with temporal, parietal, occipital, and infratentorial lesions (mean, 2.8 +/- 2.1 mm; n = 21). CONCLUSION: Laser scanning for surface registration is an accurate, robust, and easy-to-use method of patient registration for image-guided surgery. PMID- 11904032 TI - Neuronavigation by intraoperative three-dimensional ultrasound: initial experience during brain tumor resection. AB - OBJECTIVE: Three-dimensional (3-D) ultrasound is an intraoperative imaging modality used in neuronavigation as an alternative to magnetic resonance imaging (MRI). This article summarizes 4 years of clinical experience in the use of intraoperative 3-D ultrasound integrated into neuronavigation for guidance in brain tumor resection. METHODS: Patients were selected for inclusion in the study on the basis of the size and location of their lesion. Preoperative 3-D MRI data were registered and used for planning as in other conventional neuronavigation systems. Intraoperative 3-D ultrasound images were acquired three to six times, and tumor resection was guided on the basis of these updated 3-D images. RESULTS: Intraoperative 3-D ultrasound represents a good solution to the problem of brain shift in neuronavigation because it easily provides an updated, and hence more accurate, map of the patient's true anatomy in all phases of the operation. Ultrasound makes it possible to follow the progression of the operation, and it improves the radicality of tumor resection by detecting tumor tissue that would remain if the imaging technology had not been used (in 53% of the cases). Integration of 3-D ultrasound with navigation technology solves the orientation problem experienced previously with two-dimensional ultrasound in neurosurgery. The technology makes it possible to directly compare intraoperative ultrasound and MRI data regarding visualization of the lesion. Ultrasound image quality is useful for guiding surgical procedures. CONCLUSION: Intraoperative 3-D ultrasound seems to provide a time- and cost-effective way to update high-quality 3-D maps used in neuronavigation. PMID- 11904033 TI - Degloving transfacial approach with Le Fort I and nasomaxillary osteotomies: alternative transfacial approach. AB - OBJECTIVE: We present surgical results obtained with the use of an alternative transfacial approach to the central cranial base. METHODS: A degloving transfacial approach, which is a combination of the midface degloving procedure, the Le Fort I osteotomy with a pediculated cartilaginous septum, and a nasomaxillary osteotomy, was used in 13 procedures for 8 patients. The lower clivus and upper cervical spine were approached via a submucosal route, without opening of the oropharyngeal mucosa. The wall of the nasopharynx was closed with the mucosa of the bony septum. Several patients had previously undergone other surgical procedures and received radiotherapy. RESULTS: The follow-up periods ranged from 4 months to 6.4 years. The same procedure was repeated three times for one patient, with intervals of 5.5 and 1.5 months, and twice for three patients, with intervals of 8.2, 6.3, and 1.3 years. A maxillary antrotomy or bifrontal craniotomy with removal of the orbital bar was combined with this technique. No significant or insurmountable technical problems were encountered, even among patients who had undergone previous surgery or radiotherapy. CONCLUSION: Our technique is relatively simple, with good cosmetic results, and affords sufficient access to the central cranial base from the frontal base down to the upper cervical spine, especially for epidural lesions located in the midline between the carotid arteries. It offers much lower risks of damage to vital neurovascular structures, as well as of meningeal or pharyngeal infectious problems, wound dehiscence, and cerebrospinal fluid leakage. This procedure can be repeated without any increase in difficulty. PMID- 11904034 TI - Neurophysiological monitoring for the nucleus caudalis dorsal root entry zone operation. AB - OBJECTIVE: The purpose of this report is to describe a neurophysiological monitoring technique that can decrease the incidence of complications while maintaining the effectiveness of the nucleus caudalis dorsal root entry zone (DREZ) operation. METHODS: Needle electrodes were used to stimulate the supraorbital, infraorbital, mental, and median nerves after the nucleus caudalis was surgically exposed. The DREZ electrode was used to record responses from the various areas in and near the nucleus. The target site was localized. Before lesioning, the site was stimulated with the DREZ electrode and electromyographic activation was sought. If no activation was observed, a lesion was made. RESULTS: Five patients underwent a total of seven nucleus caudalis DREZ procedures with complete neurophysiological monitoring. The mean number of lesions per procedure in this series was 5.4. Six procedures (86%) resulted in immediate pain relief, and five (71%) produced persistent benefit after a mean follow-up period of 12 months. Only one patient (20%) (one of seven procedures) who underwent a unilateral DREZ procedure had ataxia, which resolved within a few days. No complications were noted at follow-up. CONCLUSION: Despite patients in this series receiving fewer lesions, the efficacy of the DREZ operation was comparable to that reported in earlier studies. There were fewer complications when neurophysiological monitoring was used. Such monitoring should be considered for nucleus caudalis DREZ operations. PMID- 11904035 TI - Meningovenous structures of the petroclival region: clinical importance for surgery and intravascular surgery. AB - OBJECTIVE: The goals of this investigation were to perform a detailed analysis of petroclival microanatomic features, to investigate the course of the abducens nerve in the petroclival region, and to identify potential causes of injury to neurovascular structures when anterior transpetrosal or transvenous endovascular approaches are used to treat pathological lesions in the petroclival region. METHODS: Petroclival microanatomic features were studied bilaterally in seven cadaveric head specimens, which were injected with colored silicone before microdissection. Another cadaveric head was used for histological section analyses. RESULTS: A lateral or medial location of the abducens nerve dural entrance porus, relative to the midline, was correlated with the course and angulation of the abducens nerve in the petroclival region. The angulation of the abducens nerve was greater and the nerve was closer to the petrous ridge in the lateral type, compared with the medial type. The abducens nerve exhibited three changes in direction, which represented the angulations in the petroclival region, at the dural entrance porus, the petrous apex, and the lateral wall of the internal carotid artery. The abducens nerve was covered by the dural sleeve and the arachnoid membrane, which became attenuated between the second and third angulation points. The abducens nerve was anastomosed with the sympathetic plexus and fixed by connective tissue extensions to the lateral wall of the internal carotid artery and the medial wall of Meckel's cave at the third angulation point. There were two types of trabeculations inside the sinuses around the petroclival region (tough and delicate). CONCLUSION: The petroclival part of the abducens nerve was protected in a dural sleeve accompanied by the arachnoid membrane. Therefore, the risk of abducens nerve injury during petrous apex resection via the anterior transpetrosal approach, with the use of the transvenous route through the inferior petrosal sinus to the cavernous sinus, should be lower than expected. The presence of two anatomic variations in the course of the abducens nerve, in addition to findings regarding nerve angulation and tethering points, may explain the relationships between adjacent structures and the susceptibility to nerve injury with either surgical or endovascular approaches. Venous anatomic variations may account for previously reported cases of subarachnoid hemorrhage with the endovascular approach. PMID- 11904036 TI - Occurrence of a glioblastoma-associated tenascin-C isoform in cerebral cavernomas and neighboring vessels. AB - OBJECTIVE: Regrowth of cerebral cavernomas after apparently complete excision, de novo occurrence, and evidence of proliferation-related patterns raise the question as to their intrinsic growth potential. A particular isoform (Type III repeat c) of the glycoprotein tenascin-C (TN-C), typically associated with the vessels of anaplastic gliomas, is regarded as a marker of vascular proliferation in lesions growing within brain tissue. This study sought to ascertain whether this isoform is expressed in cerebral cavernomas to gain further insight into the growth potential of these lesions. METHODS: Sixteen cerebral cavernomas and three fragments of normal brain underwent immunohistochemical examinations via two antibody fragments obtained by phage display technology. Previous characterization demonstrated that the fragment TN-12 recognizes the epidermal growth factor-like repeat, common to all TN-C isoforms. On the contrary, the fragment TN-11 was found to be highly specific for the Type III repeat c isoform. RESULTS: Accumulation of total TN-C was found in the vascular walls and in the interspaces between the blood cavities of all examined cavernomas. When the antibody fragment TN-11 was used, staining of the subendothelial layers occurred in both the bulk of the cavernomas and vessels of the white matter surrounding the lesions, but staining was absent in the control specimens. CONCLUSION: The distribution of the Type III repeat c isoform of TN-C, a putative marker of vascular proliferation, within cerebral cavernomas is consistent with the hypothesis of a growth potential of cerebral cavernomas. Enlargement of these lesions might involve recruitment of neighboring vasculature, which is possibly dependent on environmental conditions. PMID- 11904037 TI - Sustained arterial narrowing after prolonged exposure to perivascular endothelin. AB - OBJECTIVE: The vasoconstrictor peptide endothelin-1 (ET-1) produces narrowing of cerebral arteries and has been implicated in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage. Little is known, however, regarding the physiological consequences of prolonged exposure of arterial wall to ET-1. METHODS: In 30 rats, normal saline or 10(-8) mol/h of ET-1 was continuously applied for 3 or 5 days to the adventitial surface of the femoral artery in a Silastic cuff via an osmotic infusion pump. Vessels were examined for histopathological changes and luminal narrowing during ET-1 infusion (3 or 5 d) or at intervals from 2 to 9 days after infusion was stopped. RESULTS: Marked arterial constriction (30-40% arterial diameter reduction) was present during continuous ET-1 infusion for 3 or 5 days. For both 3- and 5-day ET-1 infusions, significant reduction in arterial cross sectional area persisted up to 4 days after cessation of infusion, after which normal caliber returned. In arteries with persistent luminal narrowing after cessation of ET-1 infusion, light microscopic findings revealed morphological changes in the vessel wall similar to those observed in cerebral vasospasm after subarachnoid hemorrhage, with apparent increased collagen deposition in media and adventitia. CONCLUSION: Continuous infusion of ET-1 produces reversible arterial narrowing that persists beyond the usual interval of physiological effect for this agent. Prolonged arterial constriction may produce physiological changes in arterial wall that act to maintain a narrowed lumen. PMID- 11904038 TI - Exposure of the dorsal root ganglion in rats to pulsed radiofrequency currents activates dorsal horn lamina I and II neurons. AB - OBJECTIVE: Application of pulsed radiofrequency (RF) currents to the dorsal ganglion has been reported to produce long-term relief of spinal pain without causing thermal ablation. The present study was undertaken to identify spinal cord neurons activated by exposure of the dorsal ganglion to pulsed RF currents in rats. METHODS: Left-sided hemilaminectomy was performed in adult Sprague Dawley rats to expose the C6 dorsal root ganglion. An RF electrode (0.5 mm diameter) with a thermocouple for temperature monitoring was positioned on the exposed ganglion, and rats were assigned to one of three treatment groups: pulsed RF treatment (20 ms of 500-kHz RF pulses delivered at a rate of 2 Hz for 120 s to produce tissue heated to 38 degrees C), continuous RF (continuous RF currents for 120 s to produce tissue heated to 38 degrees C), or sham treatment (no RF current; electrode maintained in contact with ganglion for 120 s). RESULTS: Treatment with pulsed RF but not continuous RF was associated with a significant increase in the number of cFOS-immunoreactive neurons in the superficial laminae of the dorsal horn as observed 3 hours after treatment. CONCLUSION: Exposure of the dorsal ganglion to pulsed RF currents activates pain-processing neurons in the dorsal horn. This effect is not mediated by tissue heating. PMID- 11904039 TI - Overexpression of basic fibroblast growth factor and Bcl-xL with adenoviral vectors protects primarily cultured neurons against glutamate insult. AB - OBJECTIVE: Excitatory amino acid (EAA) toxicity seems to be an important mechanism of neuronal cell death after cerebral infarction. We examined the inhibitory effects of neuronal cell death caused by EAA in vitro by means of adenoviral gene transfer of neurotrophic basic fibroblast growth factor (bFGF) and antiapoptotic Bcl-xL. METHODS: Recombinant adenoviral vectors expressing human bFGF gene with secretory signals of interleukin-2 and human Bcl-xL gene were constructed. Primarily cultured rat neuronal cells were treated with glutamate to cause EAA, and the neuroprotective effects of gene transfer by these adenoviral vectors were investigated at several time points of infection. RESULTS: Each adenoviral infection to primarily cultured neuronal cells exhibited neuroprotective effects against EAA caused by glutamate. Both gene transfer of bFGF with secretory signal and Bcl-xL transfer to neuronal cells exhibited the synergistic neuroprotective effects against EAA. These effects were most prominent with gene transfer 4 hours before glutamate insult; gene transfer performed simultaneously with and up to 4 hours after the insult exhibited definite neuroprotective effects. CONCLUSION: These experiments revealed marked neuroprotective effects of adenoviral gene transfer of bFGF and Bcl-xL into neuronal cells in vitro. The findings may lead to new approaches for treating occlusive cerebrovascular disease. PMID- 11904040 TI - History and state of neurosurgery in Austria. AB - Although surgeons in Austria, especially in Vienna, were counted among the leading specialists at the end of the 19th century, neurosurgery did not evolve as a distinct discipline before the turn of the century; achievements were episodic until Anton von Eiselsberg became an enthusiastic surgeon of the central nervous system at the beginning of the 20th century. On the threshold of modern microneurosurgery, he was succeeded in Vienna by Leopold Schonbauer and then by Herbert Kraus. Although Schonbauer kept a certain distance from neurosurgery before World War II, a special department of neurosurgery was founded at the University of Graz Medical Faculty in 1950. In contrast, it was not until 1964 that Kraus founded the first department of neurosurgery at the University of Vienna Medical Faculty, later followed by the one at Innsbruck. War injuries involving the brain during both World Wars I and II also had considerable impact on further progress in brain and spine surgery in Austria. At present, Austria harbors three university departments of neurosurgery (Vienna, Graz, and Innsbruck) and three more departments at community hospitals, in addition to four at state hospitals. Each is equipped with modern devices, including the capacity for radiosurgery in five institutions. In 1954, a scientific society was founded, but neurosurgery was not established as a distinct specialty in medicine in Austria until 1976. PMID- 11904041 TI - Healed cranial weapon injury from medieval coventry, England. AB - Archaeological excavation recovered medieval human skeletal remains from Coventry Cathedral. A disarticulated skull from the desecrated Copston Chapel displayed major cranial weapon injury. Osteological examination indicates that the injuries, although extensive, were not fatal. The cause and chronicity of the lesions are discussed. The rarity of medieval British healed cranial weapon injuries has prompted this report. Documentary sources for medieval cranial surgery are included. PMID- 11904042 TI - Cerebellar aspergillosis: case report and literature review. AB - OBJECTIVE AND IMPORTANCE: An unusual, but not unique, case of cerebellar aspergillosis associated with autologous peripheral blood stem cell transplantation for breast cancer is presented. CLINICAL PRESENTATION: A 45-year old woman with breast cancer underwent chemotherapy and radiotherapy as well as autologous peripheral blood stem cell transplantation. She developed a cerebellar aspergillosis abscess that was treated successfully with two surgical resections. INTERVENTION: After removal of pus and the abscess wall, the patient received local application of amphotericin B (AmB). She received AmB 1 mg/kg/d for 3 months and itraconazole 100 mg/kg/d for 1 year. After 3 months of AmB treatment, magnetic resonance imaging revealed that disease had not recurred. CONCLUSION: In cases of central nervous system aspergillosis, to increase the therapeutic efficiency, AmB can also be applied to the abscess cavity. Computed tomographic and contrast-enhanced magnetic resonance imaging scans play an important role in establishing early diagnosis in high-risk, immunocompromised patients. PMID- 11904044 TI - Aneurysm originating from the fenestration of the posterior cerebral artery: case report. AB - OBJECTIVE AND IMPORTANCE: A rare case of an aneurysm arising at the fenestration of the P2 segment of the posterior cerebral artery is reported. CLINICAL PRESENTATION: A 37-year-old man presented with severe headache and disturbance of consciousness. Computed tomographic scanning showed diffuse subarachnoid hemorrhage. Cerebral angiography revealed an aneurysm at the fenestration of the P2 segment of the right posterior cerebral artery. The aneurysm was located at the middle portion of the lower trunk of the fenestration. An unruptured arteriovenous malformation was incidentally found in the right thalamus. INTERVENTION: A right frontotemporal craniotomy with orbitozygomatic osteotomy was made, and the aneurysm was successfully clipped. One year after the operation, gamma knife surgery was performed for the right thalamic arteriovenous malformation. CONCLUSION: This is the first reported case of an aneurysm originating from the middle portion of a fenestrated posterior cerebral artery. PMID- 11904043 TI - Intraparenchymal tension pneumatocele complicating frontal sinus osteoma: case report. AB - OBJECTIVE AND IMPORTANCE: A relatively rare condition of intraparenchymal tension pneumatocele secondary to a frontal sinus osteoma eroding posteriorly and breaching dura mater is described. The scanty body of literature on this subject is briefly summarized, and the importance of this condition as a result of its life-threatening but readily treatable intracranial mass effect is outlined. CLINICAL PRESENTATION: The patient presented with acute deterioration in conscious state and lateralizing signs from the mass effect of gas under tension. Two weeks earlier, he had experienced vague and subtle changes in personality noticeable only to his family. INTERVENTION: The patient was cured by a frontal craniotomy, partial excision of the osteoma, and suture repair of the dural defect after evacuation of the pressurized air cavity. CONCLUSION: This rare condition should be urgently treated in the event of acute deterioration. To prevent a life-threatening situation from arising, elective surgery should be considered for patients known to have air sinus osteomas that are at risk of erosion into the cranial cavity. PMID- 11904045 TI - Intracranial aneurysm and vasculopathy after surgery and radiation therapy for craniopharyngioma: case report. AB - OBJECTIVE AND IMPORTANCE: This case report illustrates the possible occurrence of intracranial aneurysms after surgery and radiation-induced vasculopathy. CLINICAL PRESENTATION: An internal carotid bifurcation aneurysm was diagnosed in a 19-year old woman in a routine follow-up examination by magnetic resonance imaging 5 years after subtotal removal of a giant cystic craniopharyngioma treated by postoperative external radiotherapy. The presence of the aneurysm was confirmed by angiography. INTERVENTION: It was decided to treat the aneurysm by embolization with Guglielmi detachable coiling. However, at the beginning of the procedure, a few weeks after the diagnosis, a dramatic reduction in the carotid artery blood flow was observed, along with signs of thrombosis inside the aneurysm. In light of these findings, the procedure was aborted. Four months later, another angiographic examination confirmed the exclusion of the aneurysm and compensatory flow through the external carotid artery. CONCLUSION: During the assessment of patients who have undergone postoperative radiotherapy, the potential for the development of aneurysms and radiation-induced vasculopathy exists and should be kept in mind. Considering the potential for spontaneous thrombosis of these aneurysms, cautious judgment is recommended before making a decision to treat them. PMID- 11904046 TI - Trochlear nerve neuroma manifested with intractable atypical facial pain: case report. AB - OBJECTIVE AND IMPORTANCE: Trochlear nerve neuromas are extremely rare. Seventeen surgical cases of this pathological condition have been reported in the English literature. The presented case is distinct from previous reports. CLINICAL PRESENTATION: A 26-year-old woman presented with atypical facial pain. The neurological examination results were normal. Magnetic resonance imaging revealed a left parasellar mass. INTERVENTION: A left pterional craniotomy was performed, providing access to the left parasellar area. After incision of the tentorial edge, the tumor was observed to originate from the short segment of the trochlear nerve that runs between the tentorial leaves. The neuroma was totally removed. CONCLUSION: The facial pain resolved immediately after surgery. Although facial dysesthesias have been noted among patients with trochlear nerve neuromas, here the atypical facial pain was the only clinical manifestation. In all previously reported cases, neuromas originated from the cisternal segment of the trochlear nerve (always before the site of nerve entrance into the tentorial leaves) and expanded mainly into the prepontine and interpeduncular cisterns. Subtemporal and suboccipital approaches were used. In this case, the tumor arose from the short segment of the nerve running between the tentorial leaves. The tumor did not extend either into the ambient cistern or into the cavernous sinus but did involve the parasellar area. A pterional approach was appropriate for tumor removal. A trochlear nerve neuroma should be considered as a potential cause of atypical facial pain. PMID- 11904047 TI - Intraoperative assessment of aneurysm clipping using magnetic resonance angiography and diffusion-weighted imaging: technical case report. AB - OBJECTIVE AND IMPORTANCE: To use intraoperative magnetic resonance imaging, including magnetic resonance angiography and diffusion-weighted imaging, to monitor the surgical treatment of a patient with an intracranial aneurysm. TECHNIQUE: Intraoperative imaging was performed with a ceiling-mounted, mobile, 1.5-T magnet (developed in collaboration with Innovative Magnetic Resonance Imaging Systems, Inc., Winnipeg, MB, Canada) that included high-performance 20 mT/m gradients. Pre- and postclipping, intraoperative, T1-weighted, angiographic and diffusion-weighted magnetic resonance images were obtained from a patient with an incidental, 8-mm, anterior communicating artery aneurysm. RESULTS: T1 weighted images demonstrated brain anatomic features, with visible shifts induced by surgery. Magnetic resonance angiography demonstrated the aneurysm and indicated that, after clipping, the A1 and A2 anterior cerebral artery branches were patent. Diffusion-weighted studies demonstrated no evidence of brain ischemia. CONCLUSION: For the first time, intraoperative magnetic resonance imaging has been used to monitor the surgical treatment of a patient with an intracranial aneurysm. PMID- 11904048 TI - Management of intraprocedural arterial thrombus in cerebral aneurysm embolization with abciximab: technical case report. AB - OBJECTIVE AND IMPORTANCE: Thromboembolic complications after cerebral aneurysm treatment with Guglielmi detachable coils (Boston Scientific/Target, Fremont, CA) are not infrequent; in a University of California, Los Angeles institutional review of 720 treated aneurysms, thromboembolic complications occurred in 2.5% of cases. The development of intraluminal thrombus during the embolization procedure, however, may be diagnosed promptly and treated effectively with appropriate therapy. This report describes the use of intravenously administered abciximab for the treatment of intraprocedural arterial thrombus encountered during the coil embolization of a recently ruptured anterior communicating artery aneurysm. CLINICAL PRESENTATION: A 45-year-old man presented with severe headache 12 days before transfer to our institution. He had no neurological deficits at admission. Previous computed tomography of the brain demonstrated subarachnoid hemorrhage, and magnetic resonance angiography from the other institution demonstrated a 4-mm anterior communicating artery aneurysm. INTERVENTION: The patient underwent Guglielmi detachable coil embolization of the aneurysm under systemic heparinization. During the embolization, however, a thrombus developed in the proximal left A2 segment. The patient was given an intravenous infusion (20 mg) of abciximab for 10 minutes, and within 15 minutes dissolution of the thrombus was observed with no angiographic evidence of distal emboli. After reversal of general anesthesia, the patient exhibited minimal right leg weakness, which resolved within 1 hour. CONCLUSION: Abciximab may be a useful adjunct for endovascular treatment of patients with cerebral aneurysms in whom intraprocedural arterial thrombus is encountered. PMID- 11904049 TI - Interactive use of cerebral angiography and magnetoencephalography in arteriovenous malformations: technical note. AB - OBJECTIVE: To minimize the risks associated with treating cortical cerebral arteriovenous malformations (AVMs), we developed a technique combining functional imaging and cerebral angiography. The functional loci obtained by performing magnetoencephalography (MEG) are projected onto stereoscopic pairs of a stereotactically derived digital subtraction angiogram. The result is a simultaneous three-dimensional perspective of the angioarchitecture of an AVM and its relationship to the sensorimotor cortex. METHODS: Eight patients underwent multimodality brain imaging, including magnetic resonance imaging, functional mapping via MEG, and stereotactic angiography using a modified Compass fiducial system (Compass International, Rochester, MN). The coordinates derived by performing MEG were superimposed onto stereotactic, stereoscopic, angiographic pairs using custom-made distortion correction and coordinate transfer software. RESULTS: The magnetoencephalographic angiogram allowed simultaneous viewing of the angioarchitecture of the AVM nidus, the feeding vessels, and the draining veins and their relationship to the normal cerebral vasculature and functional cortex. This imaging technique was particularly valuable in identifying en passant vessels that supplied functional cortex and was used during the treatment of these lesions. CONCLUSION: The techniques of MEG and cerebral angiography were combined to provide simultaneous viewing of both modalities in a three dimensional perspective. This technique can aid in risk stratification in the management of patients with cerebral AVMs. In addition, this technique can facilitate the selective targeting of vessels, thus potentially reducing the risks associated with embolization of these formidable lesions. PMID- 11904050 TI - Transvascular coil hooking procedure to retrieve an unraveled Guglielmi detachable coil: technical note. AB - OBJECTIVE: A patient with an anterior communicating artery aneurysm was treated by use of endovascular coiling, and a Guglielmi detachable coil (Boston Scientific/Target, Fremont, CA) fractured distal to its connection to the delivering catheter. The unraveled coil floated out from the aneurysm to extend into the bifurcation of the left middle cerebral artery. We describe the microsurgical procedure used to retrieve the coil after an endovascular approach failed. METHODS: The left anterior cerebral artery was punctured just below the aneurysm neck, and a titanium microhook was introduced to anchor the coil and pull it out. Slight traction was exerted before sectioning the coil to avoid protrusion of the stump into the parent vessel. RESULTS: The unraveled coil was removed in totality without permanent morbidity. CONCLUSION: This report describes the case of a rare complication of coil embolization treated with a minimal transarterial coil hooking procedure. PMID- 11904051 TI - Overview of trauma care and its prospect. AB - Trauma is a prominent problem in modern society, which is called "the disease of developed society" or "the twin brother of modern civilization". Nowadays, over one million people die of trauma and several ten million people are injured worldwide each year. In China, over one hundred thousand people die of trauma and several millions are injured each year. Therefore, trauma has become the fifth cause of death in urban districts and the fourth cause of death in the countryside. According to the calculation of years of potential life lost (YPLL), trauma then turns into the first cause of death, indicating that the hazard to the society and loss of labor forces induced by trauma is more severe than any other diseases. Therefore, much attention has been paid to trauma research at home and abroad. This paper is to review the recent advances in trauma care and to offer some prospects for the next century's development. PMID- 11904052 TI - Hand surgery in China: advances and prospects. PMID- 11904053 TI - Ultrastructural investigation of traumatic avascular necrosis of femoral-head. AB - OBJECTIVE: To investigate ultrastructurally into traumatic avascular necrosis of femoral head. METHODS: Femoral heads were procured during endoprosthetic replacement for 8 cases of intracapsular fracture of femoral neck, which had been subjected to internal fixation and developed Stage II avascular necrosis. The specimens were processed and studied under transmission electron microscope. RESULTS: In the normal bone tissues of the avascular necrotic femoral heads, the collagen fibrils constituting the collagen bundles of the bone matrix were of uniform diameter, and were regularly arrayed with orientation. In the necrotic bone tissues of the avascular femoral heads, the collagen fibrils were of unequal diameter, and were haphazardly arrayed without orientation. From the trabeculae of the avascular femoral heads emerged three different conditions of osteocytes: (1) In the trabeculae of normal bone tissues, the osteocytes belonged to both degenerative phase and formative phase osteocytes with the latter showing signs of degeneration; (2) In the necrotic trabeculae, the osteocytes perished leaving behind empty lacunae; (3) Over the trabeculae of the interface region could be discerned many active osteogenic cells, which were on their way of transformation into osteoblasts. In the trabeculae here, the osteocytes belonged to both formative phase and degenerative phase osteocytes. CONCLUSIONS: These results suggest that in the femoral heads with traumatic avascular necrosis, there is active osteogenetic activity around the necrotic bone tissues, amply implying a local reparative response following partial bone necrosis. PMID- 11904054 TI - The effects of mild hypothermia on patients with severe traumatic brain injury. AB - OBJECTIVE: To investigate the protective effects of mild hypothermia (33-35 degrees C) on the outcome of patients with severe traumatic brain injury (TBI) (GCS<8). METHODS: Patients in the mild hypothermia group were cooled to 33-35 degrees C by cooling blanket with muscular relaxant, and patients in the normothermia group were maintained at 37-38 degrees C. RESULTS: The result showed that the mortality was 26.1% (6/23) in the mild hypothermia group and 58.3% (14/24) in the normothermia group respectively (P<0.05). The mild hypothermia also markedly reduced intracranial pressure (P<0.01 and inhibited hyperglycermia (P<0.05). No significant side-effects were found during hypothermic treatment. CONCLUSIONS: Our clinical data have demonstrated that mild hypothermia is a useful method for management of patients with severe traumatic brain injury. PMID- 11904055 TI - Protective effect of bactericidal/permeability-increasing protein in mice with E. coli sepsis. AB - OBJECTIVE: To investigate the effect of bactericidal/permeability-increasing protein(BPI) on the outcome of sepsis in mice and its possible mechanism. METHODS: Sepsis was induced by injection of 2x10(6) colony-formed unit E. coli J5 via the tail vein. BPI of 5 mg/kg or equal volume of normal saline(NS) were injected intravenously at the same time. Endotoxin and TNFalpha levels in serum were assayed using a chromogenic limulus amebocyte lysate test and ELISA respectively. RESULTS: Seventy-two hour survival rate of septic mice was significantly higher in the BPI group (15/18) than in the NS group(8/18, P<0.01). Serum endotoxin levels in the BPI group (1.3+/-0.3 and 0.7+/-0.4 &mgr;g/L) were significantly lower than those in the NS group (3.9+/-0.8 and 2.5+/-0.9 &mgr; g/L, P<0.01) 0.5 and 1 hour following injection of bacteria respectively. The peak levels of serum tumor necrosis factor-alpha(TNFalpha)in the BPI group (1.9+/ 0.6 &mgr;g/L) were also markedly lower than those in the NS group (3.8+/-0.8 &mgr;g/L, P<0.01) 1.5 hours following bacterial injection. But there was no significant difference in blood bacterial count between the BPI and NS groups 0.5, 1.5 and 3.0 hours after injection of bacteria. CONCLUSIONS: BPI has a marked protective effect on E. coli sepsis, which might be related to its action against bacterial endotoxin and its inhibition of TNFalpha production in sepsis. PMID- 11904056 TI - Experimental study of phrenic nerve transfer for treatment of brachial plexus root avulsion. AB - OBJECTIVE: To investigate the effects of neurorrhaphy, nerve grafting and nerve implantation in phrenic nerve transfer for treatment of brachial plexus root avulsion. METHODS: One hundred and eighty Sprague-Dawley rats were divided into three groups: the neurorrhaphy group, the nerve grafting group and the nerve implantation group. Evaluations such as motor nerve latency, amplitude of compound actional potentials, muscle weight, cross sectional areas of muscle fibers and myelinated axons, and muscle function were performed at 1, 2, 3, 4, 5 and 6 months postoperatively. All data were analyzed by the Student-Newman Keuls test in SAS software. RESULTS: Variable recovery in each group was found at different postoperative intervals. At six months after operation, the following results were observed in descending superiority: neurorrhaphy > nerve grafting > nerve implantation. Nerve implantation demonstrated the function recovery was 75.76%. CONCLUSIONS: This study demonstrates that neurorrhaphy is statistically superior to nerve grafting and nerve grafting superior to nerve implantation, and also confirms the validity of phrenic nerve implantation. But when no reparable distal nerve stump is available, nerve implantation is an effective method in phrenic nerve transfer for brachial plexus root avulsion to some extent. PMID- 11904057 TI - Effects of LPS, PLA(2) and OFR on proton translocation across inner mitochondrial membrane and H(+)-ATPase in the liver. AB - OBJECTIVE: To elucidate the effects of lipopolysaccharide (LPS), phospholipase A(2) (PLA(2)) and oxygen free radical (OFR) on proton transmembrane translocation and H(+)-ATPase. METHODS: The normal rats were sacrificed for preparetion liver mitochondria and submitochondrial particles for experiments in vitro. Submitochondrial particles were incubated with LPS (100 &mgr;g/mL), PLA(2) (10 u/mL) and FeSO(4)/Vit C (30/90 &mgr;mol/L) at 30 degrees C for 30 min. The proton translocation of submitochondrial particles (SMPs) were assayed with the fluorescent probe ACMA (9-amino-6-chloro-2 methoxya cridine). The mitochondria were incubated with different concentration of LPS, PLA(2) and FeSO(4)/Vit C. The H(+)-ATPase, PLA(2) and malondialdehyde (MDA) were assayed. RESULTS: The fluorescent quenching of ACMA and H(+)-ATPase activity in high dose was significantly decreased after treatment with LPS, PLA(2), FeSO(4)/Vit C (P<0.05). The mitochondrial PLA(2) activity and MDA content were significantly increased after treatment with LPS (P<0.01). CONCLUSIONS: FeSO(4)/Vit C in low dose causes increases H(+)-ATPase activity. LPS, PLA(2), FeSO(4)/Vit C might be the important factors changing H(+)-ATPase and proton translocation across the membrane. PMID- 11904058 TI - Diagnosis and treatment of post-traumatic biliary leakage. AB - OBJECTIVE: To improve the quality of management in biliary leakage following liver or bile tract injury. METHODS: All patients with liver and/or bile duct injuries from October 1987 to February 1998 inclusive were studied retrospectively in respects of their age, sex, type and mechanism of injury, the grade of liver trauma, treatment and subsequent complications. RESULTS: In 271 patients with hepatobiliary injuries, 14 (5.17%) developed a bile leak, which fell into 2 main types: Type I, injuries involving extrahepatic or first-order bile ducts (6 patients);Type II, injuries of more peripheral biliary radicles (8 patients). Most bile leakages in this series closed spontaneously in 7-14 days postoperatively. Intra-abdominal infection (28.57%) was a frequent complication which required active intervention. CONCLUSIONS: In the management of biliary leakage, it is important that (1) the leakage should be well localized; (2) adequate abdominal drainage plays a key role in controlling any type of biliary leakage; (3) decompression of the biliary tract favors the healing of injured biliary tree, especially in Type I leakage. PMID- 11904059 TI - Inhibition of T cells by direct contact with macrophages after murine-amputation injury. AB - OBJECTIVE: To determine whether macrophages post trauma have inhibitory effect on normal T cells via direct cell to cell contact. METHODS: A murine amputation injury model was used, macrophages were harvested from abdominal cavity and treated with mitomycin-C to abrogate the secretion of cytokines. Separation of T cells from splenocytes in normal mice was performed using nylon column method. Mitomycin-C treated macrophages from control and traumatized mice were added into the normal T cell culture systems, then various parameters of T cell functions were determined. RESULTS: The production and secretion of interleukin 1 (IL-1), interleukin 6 (IL-6), tumor necrosis factor (TNF) and prostaglandin E-2 (PGE-2) could be abrogated after macrophages were treated with 25 &mgr;g/mL mitomycin-C for 30 minutes. Mitomycin-C treated macrophages from traumatized mice could obviously suppress T lymphocyte transformation, IL-2 mRNA and IL-2Ralpha mRNA levels, IL-2 production, IL-2Ralpha expression, IL-2 mediated lymphocyte proliferation response of normal T cells, could not affect IL-2-IL-2R interaction but elevated suppressive action of Ts cells. Removal of Ts cells from T cells could almost abolish the inhibition of macrophages. CONCLUSIONS: Macrophages post trauma can suppress T cell functions by depressing IL-2 and IL-2Ralpha gene expression via direct cell to cell contact, and this effect may be mediated mainly by increasing the action of Ts cells. PMID- 11904060 TI - Study on early frozen section after nerve repair with "cell surgery" technique. AB - OBJECTIVE: To observe the early histological change at the nerve anastomotic sites after using "cell surgery" with conventional microsuture. METHODS: Twenty two Sprague Dawley rats were divided into three groups: the first group as the control group n=6. The left sciatic nerves were cut using conventional cut method and repaired by either two (n=2) or four (n=4) epineurial stitches of 11-0 monofilament nylon. In the second group of rats, all the experimental procedures were undertaken freezingly and trimingly with a razor blade using sawing movements (n=4) or guillotine-like movements (n=4). The nerve stumps were then brought into apposition and aligned after thawing. A drop of fibrin glue was spread over each of the sites of anastomosis under an operating microscope. For the third group of 8 rats, all the procedures were similar to those of the second group except that glue welding was done after thawing. RESULTS: After the frozen nerves were cut and thawed, the alignment of the disorganized axon became much better at anastomotic sites after fibrin glue welding. Histological sections found that the axonal alignment was good, especially at the central part of the anastomotic sites except a slight disorganization of the peripheral part. The method of glue coaptation before thawing seems to be a better method than that after thawing. CONCLUSIONS: "Cell surgery" technique can improve the nerve repair at the astomotic sites. When we weld the nerve with fibrin glue, coaptation before thawing should be achieved soon. PMID- 11904061 TI - Brain TXA(2) and PGI(2) levels after head injury with secondary insults. AB - OBJECTIVE: The brain TXA(2) and PGI(2) levels in a new rodent model of impact acceleration diffuse brain injury alone and with hypotention and hyperthermia in combination were observed to look into the relationship between TXA(2), PGI(2) levels and different types of head injury. METHODS: Thirty-two SD rats were randomized into sham, head injury alone, secondary insult alone and head injury with secondary insult groups. At 4 hours after injury or experiment, all the rats were decapitated and their brains were sampled for radioimmunoassay (RIA) measurement. RESULTS: Compared with that of sham group there were no changes in TXA(2) and PGI(2) levels in injury alone group while there was a significant augmentation in PGI(2) level in insult alone group. Both TXA(2) and PGI(2) level in injury with secondary insult group increased significantly in comparison with that of sham at 4 hours postimpact. CONCLUSIONS: PGI(2) providing energy and precursors to the injured tissue and producing some vasoactive arachidonic products, especially TXA(2), is closely connected to the severity of brain damage. PMID- 11904062 TI - Protective effects of fibronectin on vascular endothelial cells during trauma in rats. AB - OBJECTIVE: To investigate the protective effects of fibronectin (Fn) on vascular endothelial cells. METHODS: Block reticuloendothelial system model compound with injury of tail vein and femur fractures in rats were used. The level of plasma Fn and surface Fn of peritoneal macrophage cells, the numbers of circulating endothelial cells and the injured tail vein were observed. RESULTS: Fn increased the level of plasma Fn and peritoneal macrophage cells surface Fn,but decreased the number of circulating endothelial cells after infusing Fn 1.7 mg/10 g before injury. Under the scanning electron microscope, only a local focus injury to vascular endothelial cells in the rats was observed after Fn was infused, but a piecemeal injury and detachment of vascular endothelial cells were observed in rats without giving Fn. CONCLUSIONS: Fn has obvious protective effects on vascular endothelial cells in rats. PMID- 11904063 TI - Application of APACHE II scoring in ICU trauma patients. AB - OBJECTIVE: To investigate the relationship between Injury Severity Score (ISS) and Acute Physiology and Chronic Health Evaluation (APACHE II) scores and mortality rate, and to evaluate the practical significance of APACHE II in ICU trauma patients. METHODS: ISS and APACHE II scores and mortality rate of 50 ICU trauma patients were calculated by AIS-1985 revision and Knaus method, then, compared and analyzed. RESULTS: The mortality rate was directly proportional to ISS and APACHE II scores in ICU trauma patients. APACHE II was more accurate and sensitive compared to ISS. CONCLUSIONS: APACHE II is a better predictor for ICU trauma patients. ISS>=25 or APACHE II>=20 may be used as the admitting criteria for ICU trauma patients. PMID- 11904064 TI - Reconstructing opposition of thumb with microsurgical technique. PMID- 11904065 TI - Biomechanical properties of regenerated bone by mandibular distraction osteogenesis. AB - OBJECTIVE: To study the biomechanical properties of the new bone generated by mandibular distraction osteogenesis (DO). METHODS: A total of 11 healthy adult goats were randomly divided into 2 groups, the experimental group (n=9) and the control group (n=2). For the goats in the experimental group, the bilateral mandibles were gradually lengthened for 10 mm with distraction appliances. Three goats were sacrificed respectively at 2, 4 and 8 weeks after completion of distraction. Compressive, three-point bending and shearing tests were conducted on the standard regenerated bone samples and the whole unilateral mandibular specimens. For the goats in the cont rol group, no operation was made and the whole unilateral mandible was taken as the test specimen. RESULTS: The compressive strength and bending stiffness of the new bone reached the normal level at 4 and 8 weeks after completion of distraction, respectively. But the shearing strength remained significantly weaker than that of the controls at 8 weeks after distraction. CONCLUSIONS: The distraction appliance can be removed and the lengthened mandible should be exposed to adaptive functional exercise at 8 weeks after completion of distraction. PMID- 11904066 TI - Effect of burn injury on relative anaplerosis and gluconeogenesis in rats by 13C magnetic resonance spectrum. AB - OBJECTIVE: To introduce a safe and specific approach of (13)C magnetic resonance spectrum ((13)C MRS) spectroscopy and investigate the alterations in hepatic anabolism. METHODS: Relative anaplerotic, pyruvate recycling and gluconeogenic fluxes were measured by (13)C MRS isotopomer analysis of blood glucose from rats with 40% body surface area burn injury, and from rats exposed to sham injury. A short chain fatty acid, [U (13)C] propionate which was avidly extracted by the liver, was infused intravenously to deliver (13)C into the citric acid cycle. Proton-decoupled (13)C MRS of deproteinized plasma or extracts of the freeze clamped liver were used to determine the distribution of (13)C in blood or hepatic glucose. RESULTS: There was no difference in the multiplets detected in the glucose carbon-2 anomer from blood or liver after 45 or 60 minutes of the infusion of the propionate, indicating that steady-state isotopic conditions were achieved. Gluconeogenesis relative to citric acid cycle flux was not altered by burn injury; in both sham and burn groups the rate of glucose production was about equal to flux through citrate synthase. In the sham group of animals, the rate of entry of carbon skeletons into the citric acid cycle was about 4 times than that in the burn group. Similarly, flux through pyruvate kinase (again relative to citrate synthase) was significantly increased after the burn injury. CONCLUSIONS: Since results from analysis of the blood glucose are the same as that of the hepatic glucose, (13)C distribution in the glucose and hepatic metabolism can be assessed based on the (13)C MRS analysis of the blood glucose. PMID- 11904068 TI - Curative effect of wilsonii injecta on severe head injury. AB - OBJECTIVE: To study the curative effect of wilsonii injecta on severe head injury (SHI). METHODS: A total of 120 patients with SHI were divided randomly into 2 groups, the patients treated with conventional methods as Group A (n=60) and the patients treated with wilsonii injecta as Group B (n=60). The changes of neural function indexes were evaluated with Glasgow Coma Scale (GCS) before treatment and with Glasgow Outcome Scale (GOS) after treatment, simultaneously, the parameters of hemorrheological indexes (HI), brain electrical activity map (BEAM) and transcranial Doppler sonography (TCD) were observed before and after treatment. RESULTS: In Group B, the clinical GCS, the HI, the BEAM and the prognosis GOS were improved much more than those in Group A. And the TCD parameters in Group B decreased, which had significant difference compared with that in Group A (P<0.01). CONCLUSIONS: Wilsonii injecta can rapidly improve the injured p ersons' conscious states, the abnormal BEAM and the surviving quality. It suggests that the improvement of the HI is related to the relief of the vasospasm of the arterial blood vessels in the brain, which may be one of the important mechanisms of wilsonii injecta in improving the prognosis. PMID- 11904067 TI - Blocking transforming growth factor-beta receptor signaling down-regulates transforming growth factor-beta1 autoproduction in keloid fibroblasts. AB - OBJECTIVE: To study transforming growth factor-beta1 (TGF-beta1) autoproduction in keloid fibroblasts and the regulation effect of blocking TGF-beta intracellular signaling on rhTGF-beta1 autoproduction. METHODS: Keloid fibroblasts cultured in vitro were treated with either rhTGF-beta1 (5 ng/ml) or recombinant adenovirus containing a truncated type II TGF-beta receptor gene (50 pfu/cell). Their effects of regulating gene expression of TGF-beta1 and its receptor I and II were observed with Northern blot. RESULTS: rhTGF-beta1 up regulated the gene expression of TGF-beta1 and receptor I, but not receptor II. Over-expression of the truncated receptor II down-regulated the gene expression of TGF-beta1 and its receptor I, but not receptor II. CONCLUSIONS: TGF-beta1 autoproduction was observed in keloid fibroblasts. Over-expression of the truncated TGFbeta receptor II decreased TGF-beta1 autoproduction via blocking TGF beta receptor signaling. PMID- 11904069 TI - Protective effect of hemoglobin-induced heme oxygenase-1 on injured lungs caused by limb ischemia-reperfusion in rats. AB - OBJECTIVE: To determine the role of hemoglobin (HB) induced heme oxygenase-1 (HO 1) in injured lungs caused by limb ischemia-reperfusion (I/R) in rats. METHODS: A rat model of ischemia in the hind limbs was made by clamping the infrarenal aorta with a microvascular clip, and lung injury occurred after reperfusion. To induce the expression of HO-1 in the lungs, Hb was administrated intraperitoneally at 16 hours before reperfusion. Northern blotting and Western blotting were used to detect the expression of HO-1 in the lungs, and the carboxyhemoglobin (COHb) level in arterial blood was assayed. The effect of hemoglobin (Hb) on the injured lungs after limb I/R was determined by measuring the changes of lung histology, polymorphonuclear (PMN) count, malondialdehyde (MDA) content and wet-to-dry weight ratio (W/D). Zinc protoporphyrin (ZnPP), an inhibitor of HO, was used to determine whether HO-1 was induced by Hb after lung injury. RESULTS: Hb led to a significant increase in HO-1 mRNA and protein expression in the lungs, accompanied by the increase of COHb level in arterial blood. Compared with the sham controls, the lung PMN count, MDA content and W/D significantly increased at 4 hours after limb I/R, which reversed by the pretreatment with Hb at 16 hours before reperfusion. ZnPP blocked this protective role of Hb in the injured lungs. CONCLUSIONS: Hb can induce the lung HO-1 expression, which plays an important role in the defense against I/R induced lung injury in rats. PMID- 11904070 TI - Biologic characteristics of fibroblast cells cultured from the knee ligaments. AB - OBJECTIVE: To culture fibroblast cells from the knee ligaments and to study the biological characteristics of these cells. METHODS: Cells of the anterior cruciate ligament (ACL) and the medial collateral ligament (MCL) from New Zealand white rabbit were cultured in vitro. Cellular growth and expression of the collagen were analyzed. Moreover, an in vitro wound closure model was established and the healing of the ACL and the MCL cells was compared. RESULTS: Maximal growth for all these cells were obtained with Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, but RPMI 1640 and Ham's F12 media were not suitable to maintain these cells. Morphology of both ACL and MCL cells from New Zealand white rabbit was alike in vitro, but the MCL cells grew faster than the ACL cells. Both cell types produced similar amount of collagen in culture, but the ratio of collage type I to type III produced by ACL cells was higher than that produced by MCL cells. Wound closure assay showed that at 36 hours after injury, cell-free zones created in the ACL cultures were occupied partially by the ACL cells; in contrast, the wounded zone in the MCL cultures was almost completely covered by the cells. CONCLUSIONS: Although the ACL cells and the MCL cells from New Zealand white rabbit show similar appearance in morphology in culture, the cellular growth and the biochemical synthesis of collagen as well as the healing in vitro were significantly different. These differences in intrinsic properties of the two types of cells in vitro might contribute to the differential healing potentials of these ligaments in vivo. PMID- 11904071 TI - Construction of recombinant adenoviral vector Ad-CMV-hTGFbeta1 for reversion of intervertebral disc degeneration by gene transfer. AB - OBJECTIVE: To provide a highly efficient adenoviral vector Ad-CMV-hTGFbeta1 for the study of gene therapy for reversion of the intervertebral disc degeneration. METHODS: A newly developed recombinant adenoviral vector construction system was used in the study. The cDNA of hTGFbeta1 was first subcloned into a shuttle plasmid pShuttle-CMV. The resultant plasmid was linearized by digesting with restriction endonuclease PmeI, and subsequently transformed into E.coli. BJ5183 cells with an adenoviral backbone plasmid pAdEasy-1. Recombinants were selected by kanamycin resistance and confirmed by restriction endonuclease analysis. Finally, the recombinant plasmid linearized by PmeI was transfected into 293 cells. Recombinant adenoviruses were generated within 2 weeks. RESULTS: The recombinant adenoviral plasmids were cut by BamHI and PacI respectively, and the diagnostic fragments appeared in 0.8% agarose electrophoresis. The infected 293 cells showed evident cytopathic effect (CPE). The productions of PCR confirmed the presence of recombinant adenovirus. The expression of hTGFbeta1 was verified by immunohistochemical staining. CONCLUSIONS: The successful generation of the adenoviral vector Ad-CMV-hTGFbeta1 and the confirmation of the interest gene expression make it possible for the experimental study of the reversion of the intervertebral disc degeneration by gene therapy. PMID- 11904072 TI - Sequential changes of hypoxia-inducible factor 1 alpha in experimental spinal cord injury and its significance. AB - OBJECTIVE: To study the sequential changes of HIF-1alpha (hypoxia-inducible factor 1 alpha) in experimental spinal cord injury in rats and to analyze its potential effects in SCI. METHODS: A static compression model of SCI was employed in this study. Expressions of HIF-1alpha were measured with immunohistochemical staining, while flow cytometry was used to determine the apoptotic ratio and bcl 2 expressions. RESULTS: HIF-1alpha began to increase 1 day after injury, and reached the peak at 3-7 days. Two weeks later, it declined significantly. The sequential changes of HIF-1alpha coincided well with the alterations of apoptotic ratio and contents of bcl-2. CONCLUSIONS: HIF-1alpha possibly participates in the secondary ischemic and hypoxic procedures after spinal cord injury, and may mediate the traumatic apoptosis. Further understanding of HIF-1alpha may provide new therapeutic regimens for SCI. PMID- 11904073 TI - Compound injection of radix Hedysari to promote peripheral nerve regeneration in rats. AB - OBJECTIVE: To study the effect of compound injection of Radix Hedysari on peripheral nerve regeneration in rats. METHODS: Seventy-five healthy adult SD male rats, weighing 150 g, were randomized into 5 groups (15 rats in each group). The bilateral sciatic nerves of the rats were exposed and clamped with a smooth clamp to make an injury area of 2 mm. After clamp operation Group 1 was injected with compound injection of radix Hedysari (CIRH) 1.5 ml/day, Group 2 with CIRH 1.0 ml/day, Group 3 with CIRH 0.5 ml/day, Group 4 with nerve growth factor (NGF) 50 U/day, and Group 5 was taken as the control group without any management. The bilateral sciatic nerve was taken out at 3 days, 1, 2 and 4 weeks after clamping, stained with osmic acid and observed microscopically. The myelinated nerve fibers were counted. The nerve conduction velocity was determined 2 and and 4 weeks before sample taking the sciatic nerve function index was measured 4 weeks before sample taking. RESULTS: The results of nerve conduction velocity, the myelinated nerve fiber count and the sciatic functuion index in the CIRH treated groups were better than those in the control group. The results of the nerve conduction velocity and the myelinated nerve fiber count at 2 weeks and the nerve conduction velocity at 4 weeks in Group 1 were better than those of Group 4. Biological observation showed that degenerated and necrotic myelin sheath in CIRH treated Groups at 2 and 4 weeks decreased remarkably compared to the NGF treated group. CONCLUSIONS: CIRH can promote regeneration of peripheral nerves and absorption of degenerated and necrotic injured nerves. It has the same effect as NGF. PMID- 11904074 TI - Diagnosis and treatment of traumatic carotid cavernous fistula. AB - OBJECTIVE: To discuss the diagnosis and management of traumatic carotid cavernous fistula (TCCF). METHODS: In all 15 patients with TCCF confirmed by angiography, 8 patients got early diagnosis and cure. With Seldinger technique adopted in the puncture of femoral artery, Magic 3 F-1.8 F BD catheters combining with balloon were used to embolize the fistula or the internal carotid artery. RESULTS: Early diagnosis and cure were achieved in 8 patients within one week and no sequelae occurred. Seven patients with delayed diagnosis who were cured beyond one week had some sequelae such as hypopsia in 5 cases, incomplete oculomotor paralyses in 3 and incomplete abducent paralyses in 2. Among all the 15 cases, the internal carotid artery was preserved in 12 cases accounting for 80%. Occluding the fistula with sacrifice of the internal carotid artery was performed in 3 cases and no repatency of the fistula occurred by following up beyond three months. CONCLUSIONS: The preferred therapy for TCCF is to occlude the fistula using detachable balloon. The diagnosis and treatment for TCCF can significantly reduce occurrence rate of the complications and sequelae. PMID- 11904075 TI - Dynamics of cAMP/cGMP in patients under a stress state. AB - OBJECTIVE: To investigate the dynamics of plasma cAMP/cGMP in patients during cardiac surgery, and its relationship to traumatic stress. METHODS: Sixteen patients, aged 19.31 years+/-10.4 years, who underwent an open heart operation with cardiopulmonary bypass (CPB) and hypothermia were served as subjects. The arterial plasma concentrations of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were measured by radioimmunoassay 2 hours before operation, after heparinization, 20 minutes following CPB, at the end of the operation, and 24 and 72 hours postoperatively, respectively. The patients' preoperative blood samples were heparinized and the venous blood samples of 30 healthy blood donors were taken to measure the levels of cAMP and cGMP as heparin and normal controls separately. RESULTS: There were no statistical difference among the heparin control, preoperative level and normal control. The peak values of cAMP and cGMP occurred during CPB and plasma cAMP levels changed synchronously with intensities of operative stimulus to human body. However cGMP level was mainly related to the operative stimulus to the heart and CPB. The cAMP value was positively correlated with the cGMP value (r=0.6313, P<0.001). CONCLUSIONS: Dynamic variation of plasma cyclic ribonucleotide can be considered as a reference parameter for intensity of traumatic stress. PMID- 11904077 TI - Efficacy of three methods of internal fixation on femoral neck fracture. PMID- 11904076 TI - Direct detection of oxygen free radicals produced in the viscera of burned rats using electron paramagnetic resonance spectroscopy. AB - OBJECTIVE: To detect superoxide anion (O(-)(2)) signals in the heart, liver, lung and kidney tissues after burns. METHODS: Twenty-four male rats were randomized into 4 groups. The rats of experimental groups were immersed in 100 degrees C water for 15 seconds and 25% third-degree burn was created. Thoracotomy or laparotomy was performed at 5, 10, 15, 20 and 30 minutes after burn, and specimens of the heart, lung, liver kidney were obtained for burned rats. The specimens were then preserved in liquid nitrogen for cryo-preservation and detected immediately using EPR. RESULTS: The signals of superoxide O radical appeared in the heart, lung, 1iver and kidney specimens 10-15 minutes after burn. CONCLUSIONS: There is a direct evidence of oxygen free radicals (OFRs) injury to viscera of burned rats between 10-15 minutes after burn. PMID- 11904078 TI - Cerebral atrophy after acute traumatic subdural or extradural hematomas in adults. PMID- 11904080 TI - [Nosocomial infection surveillance in critically-ill patients]. PMID- 11904079 TI - A clinical study on changes in coagulation status after injuries to the extremities. PMID- 11904081 TI - [Evaluation of a new ELISA (Bartels) for detection of Legionella pneumophila antigen in urine]. AB - BACKGROUND: Detection of Legionella pneumophila soluble antigens allows rapid diagnosis of pneumonia caused by these bacteria. A new ELISA (Bartels) for antigenuria detection has recently been commercialized. We compared the new ELISA with another well-established ELISA (Binax). METHODS: To evaluate ELISA-Bartels (Legionella Urinary Antigen, Intracel, Issaquah, Washington, United States), urine samples previously characterized by ELISA Binax (Legionella Urinary Antigen Enzyme Immunoassay Kit, Binax, Portland, Maine, United States) were used. Samples came from Legionella outbreaks (n = 48), from sporadic legionellosis (n = 38), and from children with viral pneumonia (n = 21). Samples from the External Quality Control of Legionella of the European Working Group on Legionella Infections (n = 102) were also tested. Of the samples analyzed, 109 were positive in ELISA-Binax, 2 were equivocal and 98 were negative. Samples showing equivocal results were excluded from the analysis. RESULTS: The sensitivity of ELISA Bartels in comparison with that of ELISA-Binax was 98.2% (107/109) and specificity was 82.7% (81/98). In the 17 samples that were positive in ELISA Bartels and negative in ELISA-Binax, 10 were positive in ELISA-Binax after concentration by selective ultrafiltration and 6 further cases showed serology indicating or compatible with recent Legionella infection and were thus classified as true positives. CONCLUSIONS: ELISA-Bartels showed good sensitivity and specificity. Sensitivity was even higher than that of ELISA-Binax. Thus, we consider it to be an appropriate method for diagnosis of Legionella pneumonia. PMID- 11904082 TI - [Rapid diagnosis of herpetic meningoencephalitis by PCR]. AB - OBJECTIVE: To evaluate the usefulness of a rapid and simple PCR method in the diagnosis of herpetic meningoencephalitis in a pediatric population. PATIENTS AND METHODS: One hundred twenty-three cerebrospinal fluid samples from 114 pediatric patients attending the Hospital Sant Joan de Deu in Barcelona for clinical suspicion of viral meningoencephalitis or to rule out a possible herpetic etiology were evaluated. In addition to classical methods, the diagnostic technique used was PCR amplification of a highly preserved region of the DNA polymerase gene common to herpes virus 1 and 2. All patients were administered acyclovir on admission and until the results of PCR were known. If the result was negative, withdrawal of acyclovir was considered after clinical reexamination. If the result was positive, the therapy was continued for 20 days. RESULTS: Herpes simplex DNA was detected in four patients. In all patients, clinical outcome confirmed the results of PCR, whether positive or negative. PCR results were available within 6.30 and 72 hours (mean: 18 hours). CONCLUSION: This simple and rapid PCR technique can be applied in the daily routine of the microbiology laboratory. It allows early diagnosis of herpetic meningocephalitis or, when lacking, exclusion of Herpes simplex etiology. PMID- 11904083 TI - [Clinical importance of Mycobacterium kansasii and evaluation of the need to identify nontuberculous mycobacteria]. AB - BACKGROUND: The aim of this study was to evaluate the clinical importance of a single isolation of Mycobacterium kansasii in order to determine whether a single isolation of nontuberculous mycobacteria is sufficient to diagnose mycobacteriosis or whether multiple isolations are required, in which case, it would be better to wait until a second isolation is produced before undertaking identification of all nontuberculous mycobacteria. PATIENTS AND METHODS: Between 1992-1998, we studied the characteristics of 77 isolates of M. kansasii because it is the nontuberculous mycobacterium most frequently associated with disease. Our hospital has 500 beds but processed the samples from the Principe de Viana Outpatient Center and those from the district hospitals of Tudela and Estella for mycobacteria. During the study period, a mean of 3,900 specimens were processed per year. The isolates came from 22 episodes in 21 patients (19 males and 2 females). The specimens were cultured using standard techniques and confirmation was performed using gene probes (Gen-Probe, San Diego, California). RESULTS: Of the 22 episodes, positive cultures were obtained on multiple occasions in 15 (68.1%) and a single isolation was obtained in 7. All isolations came from respiratory samples. Isolations were obtained from blood on only 2 occasions (both in HIV-positive patients). Direct bacilloscopy was positive in 60% of cases with multiple isolations and negative in all cases of single isolation. Multiple isolations were found in 14 patients, 7 of whom were HIV-positive and with advanced immunodepression and 7 were HIV-negative. Of these, two were alcoholics and the remaining patients had chronic pulmonary diseases. All treated patients showed initial improvement after specific therapy and negative bacilloscopies and cultures. None of the patients with a single isolation, including two HIV positive patients, showed clinical repercussions. CONCLUSION: In our experience, a single isolation of M. kansasii was without clinical importance, even in two HIV-positive patients. Given that when the criteria of the American Thoracic Society for the diagnosis of mycobacteriosis are followed, in general, multiple mycobacteria should be isolated, a strategy of not undertaking the identification of all nontuberculous mycobacteria until a second isolation is produced can be established, unless there is serious clinical indication to suggest otherwise. PMID- 11904084 TI - [Diagnosis and epidemiological investigation of an outbreak of Clostridium perfringens food poisoning]. AB - OBJECTIVE: Clostridium perfringens is a classical agent of food-borne disease but because of the mildness and self-limiting nature of the illness, many cases are undiagnosed. This study describes the investigation of an outbreak of diarrhea due to C. perfringens in a public restaurant. METHODS: An epidemiological survey was performed and the restaurant was inspected. The specific attack rates for the items on the menu were calculated. Odds ratios were calculated to investigate the independent association between each item and the disease using a logistic regression model. Investigation of C. perfringens toxin in the feces of four symptomatic subjects and one exposed but asymptomatic subject was performed by the reverse passive latex agglutination test. RESULTS: The overall attack rate was 70.8%. The main symptoms were diarrhea (100%) and abdominal pain (94%). Significant differences were found in specific attack rates for consumption of different menu items. However, the independent contribution of each item was significant only for consumption of "ravioli with cheese sauce". Fecal detection of C. perfringens enterotoxin was positive in the four symptomatic subjects and negative in the exposed but asymptomatic subject. CONCLUSIONS: The overall attack rate in this outbreak was high. The clinical symptomatology was similar to previously published data. The epidemiological analysis revealed "ravioli with cheese sauce" to be responsible for transmission of the disease and clinical investigation together with the fecal enterotoxin detection established C. perfringens as the etiological agent. PMID- 11904085 TI - [Quinolone-resistant Neisseria gonorrhoeae: A new public health problem in Spain]. PMID- 11904086 TI - [Persistent vegetative state after meningitis due to Listeria monocytogenes]. PMID- 11904087 TI - [Subcutaneous verrucous lesions in a female renal transplant recipient]. PMID- 11904088 TI - [Probable invasive pulmonary aspergillosis and inhaled steroids treatment]. PMID- 11904089 TI - [Visceral leishmaniasis associated with human immunodeficiency virus infection (reply)]. PMID- 11904090 TI - [Isolated laringeal leishmaniasis and bone marrow culture (reply)]. PMID- 11904092 TI - [Concerning peritoneal tuberculosis]. PMID- 11904091 TI - [Osseous hydatidosis: Apropos of two cases]. PMID- 11904093 TI - [Enterococcus spp. causing infectious endocarditis. Apropos of 2 cases in natural heart valves]. PMID- 11904094 TI - [Erysipelothrix rhusiopathiae bacteremia without endocarditis or evidence of the entry point in an immunocompet woman]. PMID- 11904095 TI - [Two new cases of autochthonous hepatitis E]. PMID- 11904096 TI - [Questions and answers on the class effect]. PMID- 11904097 TI - Nonoperative treatment of splenic rupture in malaria tropica: review of literature and case report. AB - In many parts of the world malaria still is a major medical problem. Heavy international and transcontinental traveling carries malaria to non-endemic areas. Practicing physicians must be aware of the common, but also the rare and severe complications of malaria. During malaria changes in splenic structure can result in asymptomatic enlargement or complications such as hematoma formation, rupture, hypersplenism, ectopic spleen, torsion, or cyst formation. An abnormal immunological response may result in massive splenic enlargement. Spontaneous rupture of the spleen is an important and life threatening complication of Plasmodium vivax infection, but is rarely seen in Plasmodium falciparum malaria. The ability to properly diagnose and manage these complications is important. Spleen-conserving procedures should be the standard whenever possible especially in patients with a high likelihood of future exposure to malaria. PMID- 11904098 TI - Does lymphocystis occur in pacora, Plagioscion surinamensis (Sciaenidae), from Colombia? AB - A lymphocystis-like condition produced hemorrhagic lesions over the bodies of pacora Plagioscion surinamensis (Bleeker) (Perciformes: Sciaenidae) held in freshwater aquaculture in northern Colombia and increased their sensitivity to handling stress. This disease eliminated this fish from a project evaluating its aquaculture potential. Lymphocystis disease is assumed to be less damaging to the host. This condition requires more study and histological evaluation. PMID- 11904099 TI - A sero-epidemiological study of malaria in human and monkey populations in French Guiana. AB - This paper describes a sero-epidemiological study of malaria prevalence in French Guiana. An immunofluorescence assay and an enzyme-linked immunosorbent assay were used to detect antibodies against blood-stage antigens and synthetic peptides mimicking the repetitive epitope of the sporozoites of Plasmodium falciparum, Plasmodium vivax and Plasmodium malariae/brasilianum, in 218 human sera and 113 non-human primate sera collected in French Guiana. Almost all the monkey sera tested had antibodies against malaria blood-stages (98%) and a large majority (73%) also tested positive with the P. malariae/brasilianum circumsporozoite peptide. A number of primate samples also reacted positively with P. falciparum NANP repeats in a very specific manner, suggesting that monkeys in the rainforest are bitten by mosquitoes infected with human malaria parasites. Seroprevalences were lower in the humans tested but Indian tribes on the borders with Suriname and Brazil were clearly more exposed to malaria than other ethnic groups, with a prevalence of nearly 70% seropositivity. P. vivax infections accounted for much of the observed pattern of reactivity, but there was also a high frequency of positive reactions to the P. brasilianum/malariae peptide. Similarly, a large proportion of the sera obtained from Bush Negro populations tested positive for P. malariae/brasilianum repeats. These data add to the emerging evidence that non human primates might constitute a natural reservoir, not only for simian, but also for human malaria, and therefore suggest that they might be responsible for the maintenance of foci of P. malariae, and possibly of other malaria species, in isolated areas of the Amazonian rainforest. PMID- 11904100 TI - Identification of antigenically distinct populations of Leishmania (Viannia) guyanensis from Manaus, Brazil, using monoclonal antibodies. AB - Eighty Leishmania isolates from patients who contracted cutaneous leishmaniasis in the Manaus region, Amazonas State, Brazil, were identified as Leishmania (Viannia) guyanensis by the electrophoretic profiles of six enzymes. None reacted with the species-specific monoclonal antibody B19. Two L. (V.) guyanensis subpopulations were detected with the monoclonals B2 and B12. The lack of B19 expression by the L. (V.) guyanensis strains in the present study contrasts with that of the vast majority of the strains of the same parasite from eastern Amazonia and French Guyana that express the epitope. PMID- 11904102 TI - Microsatellite markers for population genetic studies in Aedes aegypti (Diptera: Culicidae) from Cote d'Ivoire: evidence for a microgeographic genetic differentiation of mosquitoes from Bouake. AB - In West Africa, Aedes aegypti (Diptera: Culicidae) (Linnaeus, C., 1762. Zweyter Theil, enhalt Beschreibungen veschiedener wichtiger Naturalien. In: Hasselquist, F. (Ed.), Reise nach Palastina in den Jahren von 1749 bis 1752, Rostock, Germany, pp. 267-606) represents the principal vector of yellow fever. This study reports the use of microsatellite markers to characterise various A. aegypti populations from Cote d'Ivoire according to a north-south transect, and to perform a temporal genetic survey of the mosquitoes. Three microsatellite loci were used to analyse individuals from four different places: Kabolo, Bouake, and two different districts of Abidjan. We found that the four populations are genetically distinct except the two Abidjan populations. In the Bouake population, the coexistence of two cryptic species, not morphologically distinguishable, seems to account for the extensive heterozygote deficiency observed. Comparison of mosquitoes from Bouake 1 year apart indicated that a dramatic change occurred in the structuring of this population over time. Taken together these results indicate that microsatellite markers could be useful for identifying various populations of A. aegypti on a microgeographic scale and to assess for temporal variation within mosquito populations. PMID- 11904101 TI - The economic loss due to treatment costs and work loss to individuals with chronic lymphatic filariasis in rural communities of Orissa, India. AB - This paper is based on 1 year round case control study to investigate the economic burden, in terms of treatment costs and loss of work to people affected with chronic lymphatic filariasis in rural communities of Orissa, Eastern India. Around three-fourths of the chronic patients have sought treatment for their condition and spent, on average an amount of INR 396 (approximately US$ 8.7) per year. The major component of the expenditure is the cost of medicines. The data on loss of work time due to chronic condition reveal that the total absenteeism to the work is significantly higher among chronic filariasis patients than controls. The total number of working hours spent per day by patients and controls are 4.94 and 6.06, respectively with a significant difference. The total absenteeism and the total number of working hours per day are influenced significantly by disease condition and other personal characteristics, namely age, sex and family type. The chronic patients lose a total of 68 days of work per year, which is equivalent to 19% of the total working time of the year. The present results show that the chronic conditions of lymphatic filariasis pose considerable burden on the patient, family and community. PMID- 11904103 TI - A comparison of two rapid field immunochromatographic tests to expert microscopy in the diagnosis of malaria. AB - In Myanmar, we tested two rapid malaria immunochromatographic kits: the OptiMAL assay for the detection of parasite lactate dehydrogenase (pLDH), and the ICT Malaria P.f./P.v. test for histidine-rich protein 2 (PfHRP2) and panmalarial antigens. A total of 229 patients were examined, of whom 133 were found to be malaria positive by Giemsa microscopy. Both OptiMAL and ICT gave lower sensitivities than previously reported. ICT sensitivity for Plasmodium falciparum and non-falciparum parasites were 86.2 and 2.9%, respectively; specificity was 76.9 and 100%, respectively. OptiMAL sensitivity for P. falciparum and non falciparum parasites were 42.6 and 47.1%, respectively; specificity was 97.0 and 96.9%, respectively. The sensitivity of both tests for the detection of both P. falciparum and non-falciparum parasites increased with parasite density. Several explanations for these results are explored. Our results raise particular concern over batch quality variations of malaria rapid diagnostic devices (MRDDs). PMID- 11904104 TI - Efficacy of artesunate and praziquantel in Schistosoma haematobium infected schoolchildren. AB - Praziquantel is the current mainstay for morbidity control of schistosomiasis. Artemisinin and its derivatives, widely used for the treatment of malaria, also display antischistosomal properties. The present study is an effort to assess the therapeutic efficacy of artesunate, an artemisinin derivative, in Schistosoma haematobium infections in a human population. The efficacy of artesunate and praziquantel were comparatively studied in primary schoolchildren from two villages, Lampsar (n=180) and Makhana (n=108), located along the Lampsar river in the delta of the Senegal River Basin in Northern Senegal (West Africa). In each village, half of the infected children were treated with a single oral dose of 40 mg/kg praziquantel and half with artesunate following the recommended malaria monotherapy regimen. For both drugs, cure and egg count reduction rates were, without apparent explanation, higher in Makhana than in Lampsar. In both villages, high and nearly comparable egg count reduction rates were obtained with both drugs at each follow-up after treatment (5, 12 and 24 weeks) in the heavy infected group of children (>50 eggs/10 ml of urine). No major adverse effects were observed. The results demonstrate that artesunate is effective against S. haematobium, but the results obtained with praziquantel were consistently better. PMID- 11904105 TI - Genetic variability of the human filarial parasite, Wuchereria bancrofti in South India. AB - The genetic variability of the lymphatic filarial parasite Wuchereria bancrofti, from three localities (one urban and two rural areas) in southern India, endemic for filariasis was studied using random amplified polymorphic DNA (RAPD) markers. The RAPD profiles were generated for 21 parasite populations (7 populations from each area), using a 10-mer random primer. The analysis of profiles indicated the existence of considerable genetic variability among parasite populations. The Nei's gene diversity between the individual populations in the 2 areas (one urban and another rural) was comparatively greater (0.3372+/-0.1462 & 0.2830+/-0.1764) than that of populations in another village (0.0490+/-0.1373). The greater genetic diversity among the former areas may be due to human migration, endemicity for long time and drug (diethyl-carbamazine citrate) pressure unlike the populations of latter village where the filariasis is relatively a recent introduction and which was never under active chemotherapy. The Nei's genetic distance was estimated and the phylogenetic tree was constructed using 'UPGMA'. These analyses indicated the prevalence of at least two genetically distinct clusters, among the populations studied, their maximum genetic distance being 0.2444. The finding of two genetic 'variants' of W. bancrofti, in the present study, may have important implications in filariasis epidemiology and control/elimination programmes. PMID- 11904106 TI - Augmented bioavailability and cysticidal activity of albendazole reformulated in soybean emulsion in mice infected with Echinococcus granulosus or Echinococcus multilocularis. AB - The anthelminthic drug, albendazole (Abz), was reformulated in a soybean oil emulsion and evaluated as a therapeutic agent for the treatment of Echinococcus granulosus and Echinococcus multilocularis in mice. Abz emulsified with 30% soybean oil (AbzE-30) resulted in higher circulating plasma concentrations of the major bioactive Abz metabolite, Abz sulfoxide (AbzSOX), after oral administration, compared with an Abz suspension. The soybean oil-emulsified Abz compound was also noted to penetrate into the hydatid cyst wall and produced higher hydatid cyst concentrations of AbzSOX. The emulsion was superior to Abz suspension in reducing the size of hydatid cysts caused by E. granulosus protoscolices collected from naturally infected sheep in Urumchi, Xinjiang Uygar Autonomous Region. In contrast, the reformulated compound's ability to reduce E. multilocularis cyst masses was only marginally superior to Abz suspension. AbzE 30 exhibited increased bioavailability and bioactivity in the treatment of murine Echinococcus hydatid cyst infections. The compound has the potential for improving therapeutic outcomes for human echinococcosis. PMID- 11904107 TI - Effects of calcium supplementation on serum lipid concentrations in normal older women: a randomized controlled trial. AB - PURPOSE: To determine the effect of supplementation with calcium citrate on circulating lipid concentrations in normal older women. SUBJECTS AND METHODS: As part of a study of the effects of calcium supplementation on fractures, we randomly assigned 223 postmenopausal women (mean [+/- SD] age, 72 +/- 4 years), who were not receiving therapy for hyperlipidemia or osteoporosis, to receive calcium (1 g/d, n = 111) or placebo (n = 112) for 1 year. Fasting serum lipid concentrations, including high-density lipoprotein (HDL) cholesterol and low density lipoprotein (LDL) cholesterol, were obtained at baseline, and at 2, 6, and 12 months. RESULTS: After 12 months, HDL cholesterol levels and the HDL cholesterol to LDL cholesterol ratio had increased more in the calcium group than in the placebo group (mean between-group differences in change from baseline: for HDL cholesterol, 0.09 mmol/L (95% confidence interval [CI]: 0.02 to 0.17; P = 0.01); for HDL/LDL cholesterol ratio, 0.05 (95% CI: 0.02 to 0.08; P = 0.001). This was largely due to a 7% increase in HDL cholesterol levels in the calcium group, with a nonsignificant 6% decline in LDL cholesterol levels. There was no significant treatment effect on triglyceride level (P = 0.48). CONCLUSION: Calcium citrate supplementation causes beneficial changes in circulating lipids in postmenopausal women. This suggests that a reappraisal of the indications for calcium supplementation is necessary, and that its cost effectiveness may have been underestimated. PMID- 11904108 TI - A 6-month randomized trial of thyroxine treatment in women with mild subclinical hypothyroidism. AB - PURPOSE: The role of thyroxine replacement in subclinical hypothyroidism remains unclear. We performed a 6-month randomized, double-blind, placebo-controlled trial to evaluate the effects of thyroxine treatment for mild subclinical hypothyroidism, defined as a serum thyroid-stimulating hormone level between 5 to 10 microU/mL with a normal serum free thyroxine level (0.8-16 ng/dL). SUBJECTS AND METHODS: We randomly assigned 40 women with mild subclinical hypothyroidism who had presented to their family practitioners to either thyroxine treatment (n = 23; 50 to 100 microg daily) or placebo (n = 17). Health-related quality of life (Hospital Anxiety and Depression scale, 30-item General Health Questionnaire), fasting lipid profiles, body weight, and resting energy expenditure were measured at baseline and 6 months. RESULTS: The most common presenting symptoms were fatigue (n = 33 [83%]) and weight gain (n = 32 [80%]). At presentation, 20 women (50%) had elevated anxiety scores and 22 (56%) had elevated scores on the General Health Questionnaire. Thirty-five women completed the study. There were no significant differences in the changes from baseline to 6 months between women in the thyroxine group and the placebo group for any of the metabolic, lipid, or anthropometric variables measured, expressed as the mean change in the thyroxine group minus the mean change in the placebo group: body mass index, -0.3 kg/m(2) (95% confidence interval [CI]: -0.9 to 0.4 kg/m(2)); resting energy expenditure, 0.2 kcal/kg/24 h (95% CI: -1.3 to 1.0 kcal/kg/24 h); and low-density lipoprotein cholesterol, -4 mg/dL (95% CI: -23 to 15 mg/dL). There was a significant worsening in anxiety scores in the thyroxine group (scores increased in 8 of 20 women and were unchanged in 2 of 20) compared with the placebo group (scores increased in 1 of 14 women and were unchanged in 6 of 14; P = 0.03). CONCLUSIONS; We observed no clinically relevant benefits from 6 months of thyroxine treatment in women with mild subclinical hypothyroidism. PMID- 11904109 TI - Water drinking as a treatment for orthostatic syndromes. AB - PURPOSE: Water drinking increases blood pressure in a substantial proportion of patients who have severe orthostatic hypotension due to autonomic failure. We tested the hypothesis that water drinking can be used as a practical treatment for patients with orthostatic and postprandial hypotension, as well as those with orthostatic tachycardia. SUBJECTS AND METHODS: We studied the effect of drinking water on seated and standing blood pressure and heart rate in 11 patients who had severe orthostatic hypotension due to autonomic failure and in 9 patients who had orthostatic tachycardia due to idiopathic orthostatic intolerance. We also tested the effect of water drinking on postprandial hypotension in 7 patients who had autonomic failure. Patients drank 480 mL of tap water at room temperature in less than 5 minutes. RESULTS: In patients with autonomic failure, mean (+/- SD) blood pressure after 1 minute of standing was 83 +/- 6/53 +/- 3.4 mm Hg at baseline, which increased to 114 +/- 30/66 +/- 18 mm Hg (P <0.01) 35 minutes after drinking. After a meal, blood pressure decreased by 43 +/- 36/20 +/- 13 mm Hg without water drinking, compared with 22 +/- 10/12 +/- 5 mm Hg with drinking (P <0.001). In patients with idiopathic orthostatic intolerance, water drinking attenuated orthostatic tachycardia (123 +/- 23 beats per minute) at baseline to 108 +/- 21 beats per minute after water drinking ( P <0.001). CONCLUSION: Water drinking elicits a rapid pressor response in patients with autonomic failure and can be used to treat orthostatic and postprandial hypotension. Water drinking moderately reduces orthostatic tachycardia in patients with idiopathic orthostatic intolerance. Thus, water drinking may serve as an adjunctive treatment in patients with impaired orthostatic tolerance. PMID- 11904111 TI - Fluconazole to prevent yeast infections in bone marrow transplantation patients: a randomized trial of high versus reduced dose, and determination of the value of maintenance therapy. AB - PURPOSE: To determine the optimal dose and duration of fluconazole antifungal prophylaxis therapy in bone marrow transplantation patients. SUBJECTS AND METHODS: Two hundred and fifty-three pediatric and adult bone marrow transplantation patients were randomly assigned to receive fluconazole 400 mg daily (high dose) or 200 mg daily (low dose) while they were neutropenic. After neutrophil recovery, patients were randomly assigned to receive maintenance therapy with either fluconazole (100 mg daily) or clotrimazole troches (10 mg 4 times daily) until 100 days after transplantation. Patients were monitored until 2 weeks after completion of early prophylaxis and to 100 days after transplantation. RESULTS: During the early prophylaxis phase, rates of yeast colonization and infections were similar in both treatment groups. By day 50, the incidence of Candida infections in the high-dose group was 4% (95% confidence interval [CI]: 1% to 7%; n = 5), compared with 1% in the low-dose fluconazole group (95% CI: 0% to 3%; n = 1; P = 0.08). During the same period, the incidence of Aspergillus infections was 4% (95% CI: 1% to 7%; n = 5) in the high-dose group and 2% (95% CI: 0% to 4%; n = 2; P = 0.33) in the low-dose group. During the maintenance prophylaxis phase, rates of yeast colonization and superficial infections were similar in the fluconazole and clotrimazole groups. Four patients developed systemic fungal infection in the maintenance phase (1 who received clotrimazole and 3 who received fluconazole). CONCLUSION: High-dose (400 mg daily) and low-dose (200 mg daily) fluconazole have similar efficacy in reducing the incidence of yeast colonization, superficial infection, and systemic infection in neutropenic pediatric and adult patients undergoing bone marrow transplantation. Rates of yeast colonization after neutrophil recovery were similar in patients treated with fluconazole or clotrimazole. PMID- 11904110 TI - Comparative effects of clonidine and dihydroergotamine on venomotor tone and orthostatic tolerance in patients with severe hypoadrenergic orthostatic hypotension. AB - PURPOSE: Clonidine, an alpha(2)-adrenergic agonist, raises blood pressure in patients with autonomic failure, in whom failure of reflex neurogenic venoconstriction leads to severe orthostatic hypotension. Because animal studies suggest that postjunctional alpha(2)-adrenoreceptors are located mainly on venous capacitance rather than arterial resistance vessels, we tested the hypothesis that venoconstriction is the main mechanism by which clonidine raises blood pressure in patients with autonomic failure. SUBJECTS AND METHODS: We measured forearm venous and arterial tone using plethysmography in 4 patients with autonomic failure before and after acute administration of clonidine (0.4 mg orally) or dihydroergotamine (0.15 mg intravenously), a known venoconstrictor agent. We also recorded supine intraarterial pressure at rest and during graded orthostatic stress with lower body negative pressure. RESULTS: Clonidine and dihydroergotamine caused similar increases in supine (mean +/- SD) arterial pressure (+23 +/- 11 mm Hg vs. and +27 +/- 5 mm Hg) and forearm vascular resistance (+36% +/- 13% vs. +28% +/- 9%). However, the drugs had different effects on forearm venous tone, which increased by 38% +/- 9% with dihydroergotamine (P = 0.01 vs. control) but was unaffected by clonidine (change = 0% +/- 14%). A single dose of clonidine was less effective than a single dose of dihydroergotamine in maintaining arterial pressure during graded orthostatic stress. CONCLUSION: In contrast with what has been hypothesized, clonidine appears to function mainly as an arterial constrictor in patients with hypoadrenergic orthostatic hypotension. Further studies are needed to determine if venoconstrictor agents are of greater therapeutic benefit in this condition than are pure arterial vasoconstrictors. PMID- 11904113 TI - Gastrointestinal involvement in polyarteritis nodosa (1986-2000): presentation and outcomes in 24 patients. AB - PURPOSE: Gastrointestinal involvement in polyarteritis nodosa carries a poor prognosis. A 1982 review from our institution reported acute abdominal syndromes in 31% of patients with polyarteritis nodosa, and that all 5 patients with acute surgical abdomens died. We reviewed our more recent experience to determine if outcomes have changed since. SUBJECTS AND METHODS: We reviewed the records of all patients with polyarteritis nodosa in our vasculitis database between 1986 and 2000. Inclusion criteria were a diagnosis of polyarteritis nodosa, symptoms or signs of gastrointestinal involvement, and either a mesenteric angiogram consistent with polyarteritis nodosa or histopathologic proof of a medium-vessel vasculitis. We calculated a prognostic (5 factor) score for all patients. RESULTS: We identified 24 patients with polyarteritis nodosa who had gastrointestinal involvement during their illness. Thirteen (54%) of the patients developed acute surgical abdomens, 3 of whom died (P = 0.02 by comparison with the historical cohort). Mean (+/- SD) prognostic scores were higher among patients in the acute abdomen group compared with those who did not have acute abdominal syndromes (1.7 +/- 0.9 vs. 0.6 +/- 0.7, P = 0.002), corresponding with the observed mortality in these groups. CONCLUSION: Gastrointestinal involvement occurs commonly in polyarteritis nodosa and carries a poor prognosis. Compared with a historical cohort at our institution, mortality from this complication may have decreased, perhaps because of earlier diagnosis. PMID- 11904114 TI - Caveat cenans! PMID- 11904112 TI - The epidemiology of Candida glabrata and Candida albicans fungemia in immunocompromised patients with cancer. AB - PURPOSE: Candida glabrata is an increasing cause of candidemia, especially at cancer and bone marrow transplant centers where fluconazole is used for antifungal prophylaxis. This yeast is less susceptible to fluconazole in vitro than is Candida albicans. We compared the characteristics of patients who had C. glabrata and C. albicans candidemia at a large cancer center. SUBJECTS AND METHODS: We searched the microbiological laboratory reports and identified 116 cases of C. glabrata candidemia between 1993 and 1999. The 116 cases of C. albicans candidemia that occurred most closely in time (before or after each case of C. glabrata candidemia) served as the control group. Data were collected from patients' medical records. RESULTS: When compared with patients who had C. albicans infection, patients with C. glabrata candidemia more often had an underlying hematologic malignancy (68 [59%] vs. 26 [22%], P = 0.0001), had an Acute Physiology and Chronic Health Evaluation (APACHE) II score > or =16 (55 [48%] vs. 28 [25%], P = 0.0002), and received fluconazole prophylaxis (57 [49%] vs. 8 [7%], P = 0.0001). Patients with C. albicans candidemia more often had concomitant infections (101 [87%] vs. 78 [67%], P = 0.0003) and septic thrombophlebitis (11 [10%] vs. 2 [2%], P = 0.01). Among patients treated with antifungal therapy, those with C. albicans candidemia had a significantly greater overall response to therapy (83/104 [80%] vs. 60/97 [62%], P = 0.005) and to primary therapy (74/104 [71%] vs. 45/97 [46%], P = 0.0003). Amphotericin B preparations were not more effective than fluconazole (19/45 [42%] vs. 20/38 [53%], P = 0.5) in patients with C. glabrata candidemia. Fluconazole was less effective against C. glabrata than against C. albicans (20/38 [53%] vs. 57/74 [77%], P = 0.008). CONCLUSION: C. glabrata has emerged as an important cause of candidemia, especially among neutropenic patients who receive fluconazole prophylaxis. PMID- 11904115 TI - Smoking cessation. AB - Smoking is a risk factor for the four leading causes of death in the United States, yet 48 million Americans--24% of the U.S. adult population--continue to smoke. Approximately 70% of people who smoke visit a physician each year, yet only half report ever being advised to quit smoking by their physician. Smoking cessation is difficult due to nicotine addiction and withdrawal symptoms. Expert groups such as the National Cancer Institute and the Agency for Health Care Policy and Research offer protocols for smoking cessation that primary care physicians can use in their office practice. Recent developments in the pharmacotherapy of smoking cessation has led the U.S. Public Health Service to update the practice guidelines for treating tobacco use and dependence. Pharmacotherapy, which includes nicotine replacement therapy, offers assistance to patients who want to stop smoking. However, the cost of pharmacotherapy may be a barrier for some. Other nonpharmacologic therapies, such as counseling, are also effective. PMID- 11904117 TI - Hypothyroidism in patients with multiple myeloma following treatment with thalidomide. PMID- 11904116 TI - Prophylactic granulocyte colony-stimulating factor in patients receiving dose intensive cancer chemotherapy: a meta-analysis. AB - PURPOSE: Several studies have evaluated the efficacy of the recombinant colony stimulating factors in reducing the severity and duration of neutropenia and the risk of infection associated with dose-intensive cancer chemotherapy. We performed a meta-analysis to define better the magnitude of this effect and to assess the generalizability of the results among different diseases and types of treatment. MATERIALS AND METHODS: We used electronic databases and citation lists to identify controlled clinical trials of the prophylactic efficacy of the colony stimulating factors on neutropenic complications. We selected randomized trials of the use of recombinant colony-stimulating factors before the onset of fever or neutropenia following systemic chemotherapy for solid tumors or malignant lymphomas. RESULTS: We identified eight controlled trials (n = 1144 patients) of prophylactic colony-stimulating factors, including five trials of filgrastim (recombinant granulocyte colony-stimulating factors) and three studies of lenograstim (glycosylated granulocyte recombinant colony-stimulating factors). Five trials were double-blind and placebo-controlled; three included untreated controls. Use of recombinant colony-stimulating factors was associated with a reduced risk of febrile neutropenia (odds ratio [OR] = 0.38; 95% confidence interval [CI]: 0.29 to 0.49), documented infection (OR = 0.51; 95% CI: 0.36 to 0.73), and infection-related mortality (OR = 0.60; 95% CI: 0.30 to 1.22), but a greater risk of bone pain (OR = 2.9; 95% CI: 1.6 to 4.8). CONCLUSION: In this meta-analysis, recombinant colony-stimulating factors were effective in reducing the risk of febrile neutropenia and documented infection associated with several malignancies and dose-intensive treatment regimens. PMID- 11904118 TI - Symptomatic dietary vitamin B(12) deficiency in a nonvegetarian population. PMID- 11904119 TI - Cases from the Osler Medical Service at Johns Hopkins University. PMID- 11904121 TI - Should mild subclinical hypothyroidism be treated? PMID- 11904120 TI - Therapeutic strategies for orthostatic intolerance: mechanisms, observations, and making patients feel better. PMID- 11904122 TI - Incidental identification of Brucellosis in a patient with a gun-shot injury. PMID- 11904123 TI - Worsening heart failure associated with COX-2 inhibitors. PMID- 11904124 TI - Arterial air embolism as a complication of pleurodesis. PMID- 11904126 TI - Why we should enhance subspecialty training: indicators of areas for program improvement. PMID- 11904125 TI - Hemodynamic monitoring during sexual intercourse and physical exercise in a patient with chronic heart failure and pulmonary hypertension. PMID- 11904127 TI - DBH and the functional taxonomy of major depressive disorder. PMID- 11904128 TI - A double blind placebo controlled trial of donepezil adjunctive treatment to risperidone for the cognitive impairment of schizophrenia. AB - BACKGROUND: Despite the beneficial effects of atypical antipsychotics on cognition, these improvements will not return most schizophrenic patients to normative standards of cognitive functioning. Therefore, other treatments need to be considered. Subtle changes in cholinergic function in schizophrenic patients provide the rationale to test the effectiveness of cholinesterase inhibitors in treating cognitive impairment in schizophrenia. METHODS: Given this, a 12-week, double-blind, placebo-controlled trial of donepezil (5 mg and 10 mg) as adjunctive treatment to risperidone was conducted in a total of 36 schizophrenic patients. RESULTS: Neither the 5-mg nor 10-mg dose of donepezil produced significant improvements in any cognitive measure compared with placebo. CONCLUSIONS: It is possible that nicotinic receptor desensitization produced by chronic tobacco use in these patients rendered their nicotinic receptors refractory to the effects of increased agonist activity produced by donepezil. An alternative treatment is the allosterically potentiating ligands, which enhance the activity of (sensitize) nicotinic receptors in the presence of acetylcholine. PMID- 11904129 TI - Genotype-controlled analysis of plasma dopamine beta-hydroxylase activity in psychotic unipolar major depression. AB - BACKGROUND: Plasma activity of dopamine beta-hydroxylase (DbetaH), the enzyme that converts dopamine to norepinephrine, is reportedly lower in patients with unipolar major depression with psychotic features (UDPF) than in those with nonpsychotic unipolar major depression (UD). Plasma DbetaH is under genetic control by the structural locus encoding DbetaH protein, DBH. This study tested the hypothesis that diagnosis-specific allelic variation at DBH accounts for lower plasma DbetaH in UDPF. METHODS: Plasma DbetaH activity was measured in samples from patients with UDPF (n = 33) and UD (n = 45). Genotypes were determined at several functional DBH polymorphisms, including C-1021T, a single nucleotide polymorphism (SNP) in the proximal 5' region that associates with variation in plasma DbetaH activity. RESULTS: Mean plasma DbetaH activity was significantly lower in UDPF than in UD. Genotyping at DBH did not reveal genetic associations distinguishing UDPF from UD. A two-way analysis of variance showed significant effects of genotype and diagnostic group but no significant interaction. CONCLUSIONS: Although the effects of the diagnosis of UDPF, and of DBH allele status, on plasma DbetaH activity were replicated, the lower plasma DbetaH in patients with UDPF was not accounted for by DBH genotype. Several explanations for this result are possible. First, other variants at DBH, or at other loci, could account for the findings. Second, nongenetic factors could account for the differences in plasma DbetaH. In this regard, we hypothesize that abnormal regulation of hypothalamic-pituitary-adrenal function in UDPF lowers expression of DbetaH protein, which could in turn alter the ratio of dopamine and norepinephrine in noradrenergic neurons, thereby promoting development of psychotic symptoms. PMID- 11904130 TI - A polymorphism in the dopamine beta-hydroxylase gene is associated with "paranoid ideation" in patients with major depression. AB - BACKGROUND: Increased dopaminergic activity may play a primary role in psychotic depression. Dopamine beta-hydroxylase (DbetaH) catalyses the key step in biosynthesis of the neurotransmitter noradrenaline from dopamine, and low DbetaH activity is a possible risk factor for developing psychotic depression. An exon 2 polymorphism (DBH*444 g/a) of the DbetaH gene (DBH) is significantly associated with both serum and cerebrospinal fluid levels of DbetaH. METHODS: We determined the genotype of the DBH*444g/a polymorphism in a cohort of 164 patients with major depression and examined the association of this polymorphism with paranoid ideation, interpersonal sensitivity, and psychoticism on the Hopkins Symptom Checklist. RESULTS: Patients who possessed the A allele were significantly more likely to have higher scores for interpersonal sensitivity and paranoia than patients without the A allele (p =.004 and p =.048, respectively), suggesting that this allele may predispose patients to paranoia in major depression. In addition, we found an association between prolactin levels in men and DBH*444 g/a genotype such that homozygous G individuals displayed significantly higher levels than homozygous A or heterozygote individuals. CONCLUSIONS: Depressed patients with the GG genotype of DbetaH have lower scores for interpersonal sensitivity and paranoid ideation. The GG genotype may be protective against the development of psychosis in the presence of a major depressive episode. PMID- 11904131 TI - Cerebellar blood volume in bipolar patients correlates with medication. AB - BACKGROUND: Cerebellar abnormalities, including decreased tissue volume, have been implicated in the pathophysiology of bipolar disorder. Relatively little research has focused on blood flow in the cerebellum of patients with bipolar disorder. Furthermore, the significance of metabolic changes in the brains of psychiatric patients may be confounded by the effects of various pharmacotherapies. Having previously found differences in cerebellar blood volume in patients with bipolar disorder compared to healthy control subjects, this study examined whether some variability in the patient population may be an effect of medication. METHODS: In this study, we have examined the association between medication status and cerebellar blood volume. Thirteen healthy comparison subjects and 21 bipolar patients underwent dynamic susceptibility contrast magnetic resonance imaging. Nine cerebellar regions were identified, and the absolute cerebellar blood volume data from each was compared to medication status measures. RESULTS: Patients on conventional antipsychotics had the lowest mean absolute blood volume measures for all cerebellar regions, whereas those on atypical antipsychotics had the highest blood volume measures. Comparison subjects had cerebellar blood volume measures in the middle, with results closer to subjects in the atypical group. CONCLUSIONS: This evidence suggests that antipsychotic treatment may influence cerebellar blood volume. This effect will be important in considering imaging studies on medicated patients with bipolar disorder and may suggest novel pathways by which these medications affect their changes. PMID- 11904132 TI - The density and spatial distribution of GABAergic neurons, labelled using calcium binding proteins, in the anterior cingulate cortex in major depressive disorder, bipolar disorder, and schizophrenia. AB - BACKGROUND: There is strong evidence for the presence of a deficit in cortical gamma aminobutyric acid (GABA) neurotransmission in schizophrenia. In this investigation we have used the calcium binding proteins (CBPs) parvalbumin (PV), calretinin (CR), and calbindin-D28K (CB) as markers of these neuronal populations, and have characterized their pattern and density in schizophrenia, bipolar disorder (BPD), and major depressive disorder (MDD). METHODS: We examined the anterior cingulate cortex (ACC) in four groups of 15 brains each from subjects with schizophrenia, MDD, and BPD, and from control subjects. Using immunocytochemistry to identify these distinct neuronal populations, we quantified their areal density and spatial pattern organization. RESULTS: There were reductions in the density of CB-labeled neurons in layer 2 in schizophrenia (34%; p =.04) and BPD (33%; p =.05) compared with control subjects; however, after correction for multiple comparisons these findings no longer attained formal statistical significance. We observed no differences in the density of the neuronal populations labeled by CR or PV in any layer of the cortex in any disorder compared with control subjects. There was increased clustering among PV labeled neurons in BPD compared with control subjects but no significant differences in the spatial organization of the other neuronal subpopulations in any disorder. CONCLUSIONS: The study provides some support for the presence of a deficit in GABAergic neurons in schizophrenia and shows that these changes are not specific to schizophrenia. The findings indicate that there may be a pathophysiological condition, shared by subjects with schizophrenia and BPD, which operates to selectively reduce the number or protein expression of CB immunoreactive neurons. PMID- 11904133 TI - Principal components of the Beck Depression Inventory and regional cerebral metabolism in unipolar and bipolar depression. AB - BACKGROUND: We determined clustering of depressive symptoms in a combined group of unipolar and patients with bipolar disorder using Principle Components Analysis of the Beck Depression Inventory. Then, comparing unipolars and bipolars, these symptom clusters were examined for interrelationships, and for relationships to regional cerebral metabolism for glucose measured by positron emission tomography. METHODS: [18F]-fluoro-deoxyglucose positron emission tomography scans and Beck Depression Inventory administered to 31 unipolars and 27 bipolars, all medication-free, mildly-to-severely depressed. BDI component and total scores were correlated with global cerebral metabolism for glucose, and voxel-by-voxel with cerebral metabolism for glucose corrected for multiple comparisons. RESULTS: In both unipolars and bipolars, the psychomotor-anhedonia symptom cluster correlated with lower absolute metabolism in right insula, claustrum, anteroventral caudate/putamen, and temporal cortex, and with higher normalized metabolism in anterior cingulate. In unipolars, the negative cognitions cluster correlated with lower absolute metabolism bilaterally in frontal poles, and in right dorsolateral frontal cortex and supracallosal cingulate. CONCLUSIONS: Psychomotor-anhedonia symptoms in unipolar and bipolar depression appear to have common, largely right-sided neural substrates, and these may be fundamental to the depressive syndrome in bipolars. In unipolars, but not bipolars, negative cognitions are associated with decreased frontal metabolism. Thus, different depressive symptom clusters may have different neural substrates in unipolars, but clusters and their substrates are convergent in bipolars. PMID- 11904134 TI - Mismatch negativity predicts psychotic experiences induced by NMDA receptor antagonist in healthy volunteers. AB - BACKGROUND: Previous studies indicate that mismatch negativity (MMN)-a preattentive auditory event-related potential (ERP)-depends on NMDA receptor (NMDAR) functioning. To explore if the strength of MMN generation reflects the functional condition of the NMDAR system in healthy volunteers, we analyzed correlations between MMN recorded before drug administration and subsequent responses to the NMDAR antagonist ketamine or the 5-HT2a agonist psilocybin. METHODS: In two separate studies, MMN was recorded to both frequency and duration deviants prior to administration of ketamine or psilocybin. Behavioral and subjective effects of ketamine and psilocybin were assessed with the Brief Psychiatric Rating Scale and the OAV Scale-a rating scale developed to measure altered states of consciousness. Correlations between ERP amplitudes (MMN, N1, and P2) and drug-induced effects were calculated in each study group and compared between them. RESULTS: Smaller MMN to both pitch and duration deviants was significantly correlated to stronger effects during ketamine, but not psilocybin administration. No significant correlations were observed for N1 and P2. CONCLUSIONS: Smaller MMN indicates a NMDAR system that is more vulnerable to disruption by the NMDAR antagonist ketamine. MMN generation appears to index the functional state of NMDAR-mediated neurotransmission even in subjects who do not demonstrate any psychopathology. PMID- 11904135 TI - Noradrenergic changes, aggressive behavior, and cognition in patients with dementia. AB - BACKGROUND: We wished to examine the integrity of the noradrenergic system in patients with Alzheimer's disease, mixed/other dementias and controls, and possible relationships between changes in the noradrenergic system and the presence of behavioral and psychiatric signs and symptoms in dementia. METHODS: Alpha(2) adrenoceptor sites were measured by radioligand binding in three cortical regions of 46 individuals with dementia and 33 elderly normal controls together with cortical noradrenaline concentration and locus coeruleus cell and neurofibrillary tangle counts. RESULTS: The alpha(2) adrenergic receptor density was unaltered in patients with Alzheimer's disease, mixed/other dementias compared with controls; however, there was a loss of locus coeruleus cells in subjects with dementia, reaching 50% within the rostral nucleus. In addition, a significant reduction was seen in the midtemporal cortical noradrenaline concentration (31% decrease) in patients with Alzheimer's disease. In subjects with dementia, there was a positive correlation between aggressive behavior and magnitude of rostral locus coeruleus cell loss, while the reduction in noradrenaline concentration correlated with cognitive impairment. CONCLUSIONS: Subgroups of patients with Alzheimer's disease may have different neurochemical changes from patients lacking these changes. Therefore, this study may have implications for the treatment of behavioral and psychiatric signs and symptoms in dementia, particularly aggressive behavior in patients with dementia. PMID- 11904136 TI - Transcranial magnetic stimulation (TMS) effects on testosterone, prolactin, and corticosterone in adult male rats. AB - BACKGROUND: Transcranial magnetic stimulation is a relatively new technique for inducing small, localized, and reversible changes in living brain tissue. Although transcranial magnetic stimulation generally results in no immediate changes in plasma corticosterone, prolactin, and testosterone, it normalizes the dexamethasone suppression test in some depressed subjects and has been shown to attenuate stress-induced increases in adrenocorticotropic hormone in rats. METHODS: In this study, serum corticosterone and testosterone concentrations were assayed in male rats immediately and 3, 6, 9, 12, 24, and 48 hours following a single transcranial magnetic stimulation or sham application. Serum prolactin concentrations were determined immediately and 2 hours following a one-time application of either transcranial magnetic stimulation or sham. RESULTS: Transcranial magnetic stimulation animals displayed significantly lower corticosterone concentrations at 6 and 24 hours following a single application compared with sham-control values. Transcranial magnetic stimulation also resulted in lower corticosterone concentrations numerically but not statistically in transcranial magnetic stimulation animals immediately after application (p =.089). No significant differences were found between groups for serum prolactin or testosterone levels at any given collection time point. CONCLUSIONS: These findings 1) suggest that transcranial magnetic stimulation alters the hypothalamic-pituitary-adrenal stress axis and 2) provide time-course data for the implications of the hormonal mechanism that may be involved in the actions of transcranial magnetic stimulation. PMID- 11904137 TI - Effects of light on low nocturnal bilirubin in winter depression: a preliminary report. AB - BACKGROUND: The light-absorbing pigments involved in the induction of treatment of winter depression are unknown. It has been proposed that circulating bilirubin serves as a photoreceptor, in part because of its similarity to the chromophore of phytochrome, a primary time-setting plant molecule. METHODS: We measured nocturnal bilirubin levels in nine patients with winter depression, and seven age and gender-matched normal comparison volunteers. RESULTS: Nocturnal bilirubin levels were lower in patients than in controls (p <.02), increased in both groups during the night (p <.0001), and increased in patients after 2 weeks of morning light treatment (p =.0009), which was accompanied by clinical improvement. CONCLUSIONS: Low nocturnal bilirubin levels may be associated with winter seasonal depression. PMID- 11904138 TI - APOE genotype and hippocampal volume change in geriatric depression. AB - BACKGROUND: Apolipoprotein E has been associated with hippocampal volume loss in Alzheimer's disease and in elderly controls. In this study, we examined the role of apolipoprotein E and hippocampal volume change in 45 older nondemented depressed subjects. All had baseline and 2-year magnetic resonance imaging scans. METHODS: Hippocampal volumes were determined using a semiautomated method. RESULTS: Compared with 36 depressed subjects without an apolipoprotein E epsilon4 allele, 9 subjects with at least one APOE epsilon4 allele showed significant decline in hippocampal volume, particularly in the right hippocampus. In a logistic model, percent change in right hippocampal volume remained associated with presence of an APOE epsilon4 allele after controlling for age, gender, Mini Mental State Examination score, and baseline total cerebral volume. CONCLUSIONS: Future studies are needed to explain this association and to determine whether the finding for the right hippocampus has a biological basis, or if it is merely a function of small sample size. PMID- 11904139 TI - Jeff's view. My other genomes. PMID- 11904140 TI - Identification of a functional site on the type I TGF-beta receptor by mutational analysis of its ectodomain. AB - Six charged amino acid residues located in the ectodomain of the full-length type I transforming growth factor (TGF)-beta receptor were individually mutated to alanine. Mutation of residues D47, D98, K102 and E104 resulted in functionally impaired receptors as demonstrated by a marked decrease in ligand-dependent signaling and ligand internalization relative to the wild-type receptor. The other two mutants (K39A and K87A) exhibited wild-type-like activity. Molecular modeling indicates that the four functionally important residues are located on the convex face of the ectodomain structure. Since mutation of these four residues affects signaling and ligand internalization but not ligand binding, we propose that this functional site is an interacting site between type I and II receptors. PMID- 11904142 TI - Activity and inhibition of plasmepsin IV, a new aspartic proteinase from the malaria parasite, Plasmodium falciparum. AB - A new aspartic proteinase from the human malaria parasite Plasmodium falciparum is able to hydrolyse human haemoglobin at a site known to be the essential primary cleavage site in the haemoglobin degradation pathway. Thus, plasmepsin IV may play a crucial role in this critical process which yields nutrients for parasite growth. Furthermore, synthetic inhibitors known to inhibit parasite growth in red cells in culture are able to inhibit the activity of this enzyme in vitro. As a result, plasmepsin IV appears to be a potential target for the development of new antiparasitic drugs. PMID- 11904141 TI - N-linked oligosaccharide chains of Sendai virus fusion protein determine the interaction with endoplasmic reticulum molecular chaperones. AB - The selectivity and individual roles of the N-linked oligosaccharide chains of Sendai virus fusion protein (F protein) in the interaction with endoplasmic reticulum molecular chaperones were investigated by analyses of transient expression of single N-glycosylation mutants and sequential immunoprecipitation. We demonstrated differential interactions depending on the location of the N linked oligosaccharide chain, and showed that these interactions were correlated with the folding and transport of F proteins. Moreover, mutant F proteins that lacked the specific N-linked oligosaccharide chains required for disulfide bond formation showed increased association with ERp57. PMID- 11904143 TI - Development of a delivery vehicle for intracellular transport of botulinum neurotoxin antagonists. AB - A targeted delivery vehicle (DV) was developed for intracellular transport of emerging botulinum neurotoxin (BoNT) antagonists. The DV consisted of the isolated heavy chain (HC) of BoNT/A coupled to a 10-kDa amino dextran via the heterobifunctional linker 3-(2-pyridylthio)-propionyl hydrazide. The HC served to target BoNT-sensitive cells and promote internalization of the complex, while the dextran served as a platform to deliver model therapeutic molecules to the targeted cells. To determine the ability of this chimeric glycoprotein to enter neurons, dextran and HC were labeled independently with the fluorescent dyes Oregon green 488 and Cy3, respectively. Internalization of DV was monitored in primary cortical cells using laser confocal microscopy. Incubation of cells for 24 h with DV resulted in discrete punctate labeling of both soma and processes. The Cy3 and Oregon green 488 signals were generally co-localized, suggesting that the complex remained in the same intracellular compartment during the initial 24 h. The DV-associated fluorescence was reduced progressively by co-application of increasing concentrations of unlabeled BoNT/A holotoxin. The results suggest that the BoNT/A HC is able to mediate internalization of a coupled dextran, even though the latter bears no resemblance to the BoNT/A light chain (LC). The HC of BoNT/A thus offers promise as a selective carrier to deliver BoNT antagonists to the nerve terminal cytoplasm for inhibiting the proteolytic activity of internalized BoNT/A LC. PMID- 11904145 TI - Sucrose is involved in the diazotrophic metabolism of the heterocyst-forming cyanobacterium Anabaena sp. AB - Sucrose synthase (SuS) expression was studied in the filamentous, nitrogen-fixing cyanobacterium Anabaena sp. PCC 7119. SuS activity, SusA polypeptide, and susA mRNA levels were lower in cells cultured diazotrophically than in the presence of combined nitrogen. An insertional susA mutant presented a dramatic increase in sucrose levels, whereas the disaccharide was not detectable in a susA overexpressing strain, indicating that SusA is involved in the cleavage of sucrose in vivo. Diazotrophic growth was impaired in the susA overexpressing strain, suggesting a role for sucrose in diazotrophic metabolism and the involvement of SusA in the control of carbon flux in the N(2)-fixing filament. PMID- 11904144 TI - Chickens' Cry2: molecular analysis of an avian cryptochrome in retinal and pineal photoreceptors. AB - We have identified and characterized an ortholog of the putative mammalian clock gene cryptochrome 2 (Cry2) in the chicken, Gallus domesticus. Northern blot analysis of gCry2 mRNA indicates widespread distribution in central nervous and peripheral tissues, with very high expression in pineal and retina. In situ hybridization of chick brain and retina reveals expression in photoreceptors and in visual and circadian system structures. Expression is rhythmic; mRNA levels predominate in late subjective night. The present data suggests that gCry2 is a candidate avian clock gene and/or photopigment and set the stage for functional studies of gCry2. PMID- 11904146 TI - ADP-evoked phospholipase C stimulation and adenylyl cyclase inhibition in glioma C6 cells occur through two distinct nucleotide receptors, P2Y(1) and P2Y(12). AB - In this study we characterized the subtypes of nucleotide P2Y receptors that respond to ADP in glioma C6 cells. Direct visualization of phosphatidylinositol 4,5-bisphosphate at the cell surface revealed that extracellular ADP activates phospholipase C (PLC). Knock-down of P2Y(1) receptor with antisense oligonucleotide, as well as treatment with MRS2179 and pyridoxal-phosphate-6 azophenyl-2',4'-disulfonic acid (P2Y(1) antagonists), attenuates receptor mediated PLC activity. Adenylyl cyclase inhibition by ADP remains unchanged under these conditions. Reverse transcription-PCR analysis showed that P2Y(12) receptor is expressed in C6 cells. We therefore conclude that, in glioma C6 cells, two P2Y receptor subtypes are present: P2Y(1), coupled to PLC, and P2Y(12), negatively coupled to adenylyl cyclase. PMID- 11904147 TI - Caspase 3 regulates phosphatidylserine externalization and phagocytosis of oxidatively stressed erythrocytes. AB - The appearance of phosphatidylserine (PS) on the outer surface of red cells is an important signal for their uptake by macrophages. We report for the first time that procaspase 3 present in the anucleated mature human erythrocyte is activated under oxidative stress induced by t-butylhydroperoxide leading to impairment of the aminophospholipid translocase, PS externalization and increased erythrophagocytosis. This is the first report linking caspase 3 activation to inhibition of flippase activity and uptake of red cells by macrophages. PMID- 11904148 TI - Glucose induces and leptin decreases expression of uncoupling protein-2 mRNA in human islets. AB - Elevated islet uncoupling protein-2 (UCP-2) impairs beta-cell function and UCP-2 may be increased in clinical obesity and diabetes. We investigated the effects of glucose and leptin on UCP-2 expression in isolated human islets. Human islets were incubated for 24 h with glucose (5.5-22 mmol/l)+/-leptin (0-10 nmol/l). Some islet batches were incubated at high (22 mmol/l), and subsequently lower (5.5 mmol/l), glucose to assess reversibility of effects. Leptin effects on insulin release were also measured. Glucose dose-dependently increased UCP-2 expression in all islet batches, maximally by three-fold. This was not fully reversed by subsequently reduced glucose levels. Leptin decreased UCP-2 expression by up to 75%, and maximally inhibited insulin release by 47%, at 22 mmol/l glucose. This is the first report of UCP-2 expression in human islets and provides novel evidence of its role in the loss of beta-cell function in diabetes. PMID- 11904149 TI - Starvation-induced degradation of yeast hexose transporter Hxt7p is dependent on endocytosis, autophagy and the terminal sequences of the permease. AB - The yeast high-affinity glucose transporters Hxt6p and Hxt7p are rapidly degraded during nitrogen starvation in the presence of high concentrations of fermentable carbon sources. Our results suggest that degradation is mainly due to the stimulation of general protein turnover and not caused by a mechanism specifically triggered by glucose. Analysis of Hxt6p/7p stability and cellular distribution in end4, aut2 and apg1 mutants indicates that Hxt7p is internalized by endocytosis, and autophagy is involved in the final delivery of Hxt7p to the vacuole for proteolytic degradation. Internalization and degradation of Hxt7p were blocked after truncation of its N-terminal hydrophilic domain. Nevertheless, this fully functional and stabilized hexose transporter could not maintain fermentation capacity of the yeast cells under starvation conditions, indicating a regulatory constraint on glucose uptake. PMID- 11904150 TI - Cooperative activity of phospholipid-N-methyltransferases localized in different membranes. AB - The possibility that the phospholipid-N-methyltransferases from yeast are capable of acting upon a phospholipid substrate, localized in a different membrane than in which the enzymes reside ('trans-catalysis' hypothesis), was investigated using cho2 and opi3 gene disruptant strains, which are defective in phosphatidylethanolamine transferase (PEMT) and phospholipid methyltransferase (PLMT), respectively. When cell homogenates or microsomes of the two disruptant strains are mixed, the combined methyltransferase activity, measured as the incorporation of [(3)H]methyl label from S-adenosyl methionine, exceeds that expected based on the separate activities of PEMT and PLMT. The increased incorporation implies that monomethylphosphatidylethanolamine generated by PEMT becomes available for PLMT, as evidenced by increased synthesis of dimethylphosphatidylethanolamine and phosphatidylcholine. The kinetics of the cooperativity suggest a collision-based process, enabling either transport of substrate or 'trans-catalysis'. PMID- 11904151 TI - NF-kappaB elements contribute to junB inducibility by lipopolysaccharide in the murine macrophage cell line RAW264.7. AB - Macrophages respond to bacterial lipopolysaccharide (LPS) by activating latent cis-acting factors that initiate transcription of immediate early genes. One such immediate early gene, junB, is induced by LPS in macrophages within 30 min. To identify elements that mediate the induction of junB by LPS, upstream and downstream sequences flanking the junB gene were examined by transient expression in the RAW264.7 murine macrophage cell line using a luciferase reporter gene vector containing the junB minimal promoter. A >10-fold enhancement was associated with a 222 bp region downstream of the junB promoter in response to LPS. Transient reporter assays demonstrated that multiple nuclear factor (NF) kappaB sites are required for inducibility of junB by LPS in RAW264.7 cells. Electrophoretic mobility shift assays confirmed binding of LPS-induced nuclear proteins included p50/p65 heterodimers at these NF-kappaB sites. PMID- 11904152 TI - Induction of inducible nitric oxide synthase expression by 18beta-glycyrrhetinic acid in macrophages. AB - Glycyrrhizin (GL), a triterpenoid saponin fraction of licorice, is reported to have anti-viral and anti-tumor activities and is metabolized to 18beta glycyrrhetinic acid (GA) in the intestine by intestinal bacteria. However, the mechanism underlying its effects is poorly understood. To further elucidate the mechanism of GA, the aglycone of GL, we investigated the effects of GA on the release of nitric oxide (NO) and at the level of inducible NO synthase (iNOS) gene expression in mouse macrophages. We found that GA elicited a dose-dependent increase in NO production and in the level of iNOS mRNA. Since iNOS transcription has been shown to be under the control of the transcription factor nuclear factor kappaB (NF-kappaB), the effects of GA on NF-kappaB activation were examined. Transient expression assays with NF-kappaB binding sites linked to the luciferase gene revealed that the increased level of iNOS mRNA, induced by GA, was mediated by the NF-kappaB transcription factor complex. By using DNA fragments containing the NF-kappaB binding sequence, GA was shown to activate the protein/DNA binding of NF-kappaB to its cognate site, as measured by electrophoretic mobility shift assay. These results demonstrate that GA stimulates NO production and is able to up-regulate iNOS expression through NF-kappaB transactivation in macrophages. PMID- 11904153 TI - Conformational change and destabilization of cataract gammaC-crystallin T5P mutant. AB - Human lens gammaC-crystallin and T5P mutant were cloned, and their biophysical properties and thermodynamic stability were studied. CRYGC (T5P) is one of the many gamma-crystallin mutant genes for autosomal dominant congenital cataracts. This mutation is associated with Coppock-like cataract, and has the phenotype of a dust-like opacity of the fetal lens nucleus. During cloning and overexpression, the majority of T5P mutant was found in the inclusion body. This property is unique among the many cataract gamma-crystallin mutant genes. It is thus worthwhile to study what factors contribute to this unique property of gammaC crystallin. One possibility is changes in conformation and stability, which can be studied using spectroscopic measurements. In this study, conformational change was studied by circular dichroism and fluorescence measurements, and conformational stability was determined by thermal unfolding probed by Trp fluorescence and time-dependent light scattering. The T5P mutation obviously changes conformation and decreases conformational stability. PMID- 11904154 TI - Novel approach reveals localisation and assembly pathway of the PsbS and PsbW proteins into the photosystem II dimer. AB - A blue-native gel electrophoresis system was combined with an in organello import assay to specifically analyse the location and assembly of two nuclear-encoded photosystem II (PSII) subunits. With this method we were able to show that initially the low molecular mass PsbW protein is not associated with the monomeric form of PSII. Instead a proportion of newly imported PsbW is directly assembled in dimeric PSII supercomplexes with very fast kinetics; its negatively charged N-terminal domain is essential for this process. The chlorophyll-binding PsbS protein, which is involved in energy dissipation, is first detected in the monomeric PSII subcomplexes, and only at later time points in the dimeric form of PSII. It seems to be bound tighter to the PSII core complex than to light harvesting complex II. These data point to radically different assembly pathways for different PSII subunits. PMID- 11904155 TI - Evidence for an operative glyoxylate cycle in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius. AB - Both key enzymes for the glyoxylate cycle, isocitrate lyase (EC 4.1.3.1) and malate synthase (EC 4.1.3.2), were purified and characterized from the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius. Whereas the former enzyme was copurified with the aconitase, the latter enzyme could be enriched to apparent homogeneity. Amino acid sequencing of three internal peptides of the isocitrate lyase revealed the presence of highly conserved residues. With respect to cofactor requirement and quarternary structure the crenarchaeal malate synthase might represent a novel type of this enzyme family. High activities of both glyoxylate cycle enzymes could already be detected in extracts of glucose grown cells and both increased about two-fold in extracts of acetate grown cells. PMID- 11904156 TI - Identification of genes expressed preferentially in the honeybee mushroom bodies by combination of differential display and cDNA microarray. AB - To clarify the molecular basis underlying the neural function of the honeybee mushroom bodies (MBs), we identified three genes preferentially expressed in MB using cDNA microarrays containing 480 differential display-positive candidate cDNAs expressed locally or differentially, dependent on caste/aggressive behavior in the honeybee brain. One of the cDNAs encodes a putative type I inositol 1,4,5 trisphosphate (IP(3)) 5-phosphatase and was expressed preferentially in one of two types of intrinsic MB neurons, the large-type Kenyon cells, suggesting that IP(3)-mediated Ca(2+) signaling is enhanced in these neurons. PMID- 11904157 TI - Molecular basis for exaggerated sensitivity to mexiletine in the cardiac isoform of the fast Na channel. AB - Cardiac sodium channels have been shown to have a higher sensitivity to local anesthetic agents, such as lidocaine, than the sodium channels of other tissues. To examine if this is also true for mexiletine, we have systematically measured mexiletine sensitivity of the Na channel isoforms, rH1, (mu)1, and rBII, which were transiently expressed in human embryonic kidney (HEK) 293 cells. We confirmed that the cardiac isoform rH1 exhibited the highest sensitivity among the three tested channel isoforms. In rH1, (mu)1, and rBII, the respective IC(50) values were 62, 294, and 308 microM mexiletine, in regard to tonic block, and 18, 54, and 268 microM mexiletine, in relation to use (8 Hz)-dependent block. The relatively high drug sensitivity of rH1 was an invariant finding, irrespective of channel state or whether channels were subjected to infrequent or frequent depolarizing stimuli. Mutating specific amino acids in the skeletal muscle isoform (mu)1 (namely, (mu)1-I433V and (mu)1-S251A) to those of the cardiac isoform at putative binding sites for local anesthetic agents revealed that only one of the point mutations ((mu)1-S251A) has relevance to the high cardiac drug sensitivity, because mexiletine produced significantly more use-dependent and tonic block in (mu)1-S251A than wild-type (mu)1. PMID- 11904158 TI - Cadherin-like receptor binding facilitates proteolytic cleavage of helix alpha-1 in domain I and oligomer pre-pore formation of Bacillus thuringiensis Cry1Ab toxin. AB - Cry toxins form lytic pores in the insect midgut cells. The role of receptor interaction in the process of protoxin activation was analyzed. Incubation of Cry1Ab protoxin with a single chain antibody that mimics the cadherin-like receptor and treatment with Manduca sexta midgut juice or trypsin, resulted in toxin preparations with high pore-forming activity in vitro. This activity correlates with the formation of a 250 kDa oligomer that lacks the helix alpha-1 of domain I. The oligomer, in contrast with the 60 kDa monomer, was capable of membrane insertion as judged by 8-anilino-1-naphthalenesulfonate binding. Cry1Ab protoxin was also activated to a 250 kDa oligomer by incubation with brush border membrane vesicles, presumably by the action of a membrane-associated protease. Finally, a model where receptor binding allows the efficient cleavage of alpha-1 and formation of a pre-pore oligomeric structure that is efficient in pore formation, is presented. PMID- 11904160 TI - The P2X(7) receptor from Xenopus laevis: formation of a large pore in Xenopus oocytes. AB - The purinergic P2X(7) receptor is an ATP-receptor channel predominantly expressed in immune cells. P2X(7) has been cloned from human, rat and mouse. Here we report cloning of the Xenopus laevis P2X(7) receptor (xP2X(7)). xP2X(7) is only about 50% identical to the mammalian homologues, shows a broad tissue expression pattern, and has the electrophysiological characteristics typical of a P2X(7) receptor: low agonist affinity (EC(50) about 2.6 mM) and a non-desensitizing current. Moreover, expression of xP2X(7) in Xenopus oocytes is sufficient to induce the formation of a large pore, which is permeable to large cations such as NMDG(+). Identification of a non-mammalian P2X(7) receptor may help to identify functionally important parts of the protein. PMID- 11904159 TI - Voltage-dependent transient currents of human and rat 5-HT transporters (SERT) are blocked by HEPES and ion channel ligands. AB - The hyperpolarization-activated transient current of mammalian 5 hydroxytryptamine transporters (SERT) expressed in Xenopus oocytes was studied. Human (h) and rat (r) SERT transient currents are blocked by HEPES with changes in the waveform kinetics, and the blockade of hSERT has use-dependent properties. HEPES also changes the time course of the prepriming step, especially for hSERT. Transient currents at hSERT and rSERT are also blocked by spermine and spermidine in the mM range, and by fluoxetine, cocaine, QX-314, and QX-222 in the microM range. These pharmacological and kinetic properties of transient current blockade emphasize the similarities between the transient current and phenomena at ion channels. PMID- 11904161 TI - Clusterin is an extracellular chaperone that specifically interacts with slowly aggregating proteins on their off-folding pathway. AB - Clusterin is an extracellular mammalian chaperone protein which inhibits stress induced precipitation of many different proteins. The conformational state(s) of proteins that interact with clusterin and the stage(s) along the folding and off folding (precipitation-bound) pathways where this interaction occurs were previously unknown. We investigated this by examining the interactions of clusterin with different structural forms of alpha-lactalbumin, gamma-crystallin and lysozyme. When assessed by ELISA and native gel electrophoresis, clusterin did not bind to various stable, intermediately folded states of alpha-lactalbumin nor to the native form of this protein, but did bind to and inhibit the slow precipitation of reduced alpha-lactalbumin. Reduction-induced changes in the conformation of alpha-lactalbumin, in the absence and presence of clusterin, were monitored by real-time (1)H NMR spectroscopy. In the absence of clusterin, an intermediately folded form of alpha-lactalbumin, with some secondary structure but lacking tertiary structure, aggregated and precipitated. In the presence of clusterin, this form of alpha-lactalbumin was stabilised in a non-aggregated state, possibly via transient interactions with clusterin prior to complexation. Additional experiments demonstrated that clusterin potently inhibited the slow precipitation, but did not inhibit the rapid precipitation, of lysozyme and gamma crystallin induced by different stresses. These results suggest that clusterin interacts with and stabilises slowly aggregating proteins but is unable to stabilise rapidly aggregating proteins. Collectively, our results suggest that during its chaperone action, clusterin preferentially recognises partly folded protein intermediates that are slowly aggregating whilst venturing along their irreversible off-folding pathway towards a precipitated protein. PMID- 11904163 TI - NMR studies of flexible peptides in cavities mimicking the synaptic cleft. AB - The interaction of neuropeptides with post-synaptic receptors is characterised by a high entropic barrier originating from the combination of nanomolar concentration with low conformer population. The influence of high viscosity environments on conformer distribution can help overcome this difficulty. In an attempt to simulate the physicochemical conditions of the synaptic cleft, (15)N labelled enkephalin has been studied in polyacrylamide gels swollen by different aqueous solutions in the temperature range 273-293 K. Nuclear Overhauser enhancement spectra in the gel pores are consistent with a conformational selection or a slowing down of internal motions that can favour the interaction of the peptide with the receptor. PMID- 11904162 TI - Solution structure of polyglutamine tracts in GST-polyglutamine fusion proteins. AB - Aggregation of expanded polyglutamine (polyQ) seems to be the cause of various genetic neurodegenerative diseases. Relatively little is known as yet about the polyQ structure and the mechanism that induces aggregation. We have characterised the solution structure of polyQ in a proteic context using a model system based on glutathione S-transferase fusion proteins. A wide range of biophysical techniques was applied. For the first time, nuclear magnetic resonance was used to observe directly and selectively the conformation of polyQ in the pathological range. We demonstrate that, in solution, polyQs are in a random coil conformation. However, under destabilising conditions, their aggregation behaviour is determined by the polyQ length. PMID- 11904164 TI - Cysteine-scanning mutagenesis study of the sixth transmembrane segment of the Na,K-ATPase alpha subunit. AB - The accessibility of the residues of the sixth transmembrane segment (TM) of the Bufo marinus Na,K-ATPase alpha subunit was explored by cysteine scanning mutagenesis. Methanethiosulfonate reagents reached only the two most extracellular positions (T803, D804) in the native conformation of the Na,K-pump. Palytoxin induced a conductance in all mutants, including D811C, T814C and D815C which showed no active electrogenic transport. After palytoxin treatment, four additional positions (V805, L808, D811 and M816) became accessible to the sulfhydryl reagent. We conclude that one side of the sixth TM helix forms a wall of the palytoxin-induced channel pore and, probably, of the cation pathway from the extracellular side to one of their binding sites. PMID- 11904165 TI - Transforming growth factor-beta1 induces collagen synthesis and accumulation via p38 mitogen-activated protein kinase (MAPK) pathway in cultured L(6)E(9) myoblasts. AB - Transforming growth factor-beta (TGF-beta) plays a pivotal role in the extracellular matrix accumulation observed in chronic progressive tissue fibrosis, but the intracellular signaling mechanism by which TGF-beta stimulates this process remains poorly understood. We examined whether mitogen-activated protein kinase (MAPK) routes were involved in TGF-beta1-induced collagen expression in L(6)E(9) myoblasts. TGF-beta1 induced p38 and extracellular signal regulated kinase (ERK) 1/2 phosphorylation whereas no effect on Jun N-terminal kinase phosphorylation was observed. Biochemical blockade of p38 but not of the ERK MAPK pathway abolished TGF-beta1-induced alpha(2)(I) collagen mRNA expression and accumulation. These data indicate that TGF-beta1-induced p38 activation is involved in TGF-beta1-stimulated collagen synthesis. PMID- 11904166 TI - The interaction of the bisphosphorylated N-terminal arm of cardiac troponin I-A 31P-NMR study. AB - Cardiac troponin I, the inhibitory subunit of the heterotrimeric cardiac troponin (cTn) complex is phosphorylated by protein kinase A at two serine residues located in its heart-specific N-terminal extension. This flexible arm interacts at different sites within cTn dependent on its phosphorylation degree. Bisphosphorylation is known to induce conformational changes within cTnI which finally lead to a reduction of the calcium affinity of cTnC. However, as we show here, the bisphosphorylated cTnI arm does not interact with cTnC, but with cTnT and/or cTnI. PMID- 11904167 TI - Noladin ether, a putative novel endocannabinoid: inactivation mechanisms and a sensitive method for its quantification in rat tissues. AB - The occurrence of the novel proposed endocannabinoid, noladin ether (2 arachidonyl glyceryl ether, 2-AGE) in various rat organs and brain regions, and its inactivation by intact C6 glioma cells, were studied. 2-AGE was measured by isotope dilution liquid chromatography-atmospheric pressure chemical ionization mass spectrometry, with a detection limit of 100 fmol. A compound with the same mass and chromatographic/chemical properties as 2-AGE was found in whole brain, with the highest amounts in the thalamus and hippocampus. Synthetic [(3)H]2-AGE was inactivated by intact rat C6 glioma cells by a time- and temperature dependent process consisting of cellular uptake and partial incorporation into phospholipids. Further data suggested that 2-AGE is taken up by cells via the anandamide/2-arachidonoyl glycerol (2-AG) membrane transporter(s), and biosynthesized in a different way as compared to 2-AG. PMID- 11904168 TI - Purification and characterisation of the BIOH protein from the biotin biosynthetic pathway. AB - Conversion of pimeloyl-coenzyme A (CoA) to biotin in Escherichia coli requires at least four enzymes encoded by genes in the bio operon. One gene, bioH, which is not present in the bioABFCD operon, is required for the synthesis of pimeloyl-CoA but its exact role in formation of this intermediate is unknown. To investigate this further, we have overexpressed and purified the bioH gene products from both E. coli (BIOH EC) and Neisseria meningitis (BIOH NM) in E. coli. When purified BIOH was incubated with excess CoA and analysed by electrospray mass spectrometry a species of mass corresponding to a BIOH:CoA complex was observed. Mutation of a conserved serine residue to alanine (BIOH EC S82A) did not prevent CoA binding. This is the first report of the purification of BIOH and the observation of a small molecule bound to the protein provides clues to its role in pimeloyl-CoA synthesis. PMID- 11904169 TI - Pin1 modulates the dephosphorylation of the RNA polymerase II C-terminal domain by yeast Fcp1. AB - The reversible phosphorylation of serine and threonine residues N-terminal to proline (pSer/Thr-Pro) is an important signaling mechanism in the cell. The pSer/Thr-Pro moiety exists in the two distinct cis and trans conformations, whose conversion is catalyzed by the peptidyl-prolyl isomerase (PPIase) Pin1. Among others, Pin1 binds to the phosphorylated C-terminal domain (CTD) of the largest subunit of the RNA polymerase II, but the biochemical and functional relevance of this interaction is unknown. Here we confirm that the CTD phosphatase Fcp1 can suppress a Pin1 mutation in yeast. Furthermore, this genetic interaction requires the phosphatase domain as well as the BRCT domain of Fcp1, suggesting a critical role of the Fcp1 localization. Based on these observations, we developed a new in vitro assay to analyze the CTD dephosphorylation by Fcp1 that uses only recombinant proteins and mimics the in vivo situation. This assay allows us to present strong evidence that Pin1 is able to stimulate CTD dephosphorylation by Fcp1 in vitro, and that this stimulation depends on Pin1's PPIase activity. Finally, Pin1 significantly increased the dephosphorylation of the CTD on the Ser(5)-Pro motif, but not on Ser(2)-Pro in yeast, which can be explained with Pin1's substrate specificity. Together, our results indicate a new role for Pin1 in the regulation of CTD phosphorylation and present a further example for prolyl isomerization-dependent protein dephosphorylation. PMID- 11904172 TI - Ribosome as a molecular machine. AB - General principles of structure and function of the ribosome are surveyed, and the translating ribosome is regarded as a molecular conveying machine. Two coupled conveying processes, the passing of compact tRNA globules and the drawing of linear mRNA chain through intraribosomal channel, are considered driven by discrete acts of translocation during translation. Instead of mechanical transmission mechanisms and power-stroke 'motors', thermal motion and chemically induced changes in affinities of ribosomal binding sites for their ligands (tRNAs, mRNA, elongation factors) are proposed to underlie all the directional movements within the ribosomal complex. The GTP-dependent catalysis of conformational transitions by elongation factors during translation is also discussed. PMID- 11904173 TI - Translocation of tRNA during protein synthesis. AB - Coupled translocation of tRNA and mRNA in the ribosome during protein synthesis is one of the most challenging and intriguing problems in the field of translation. We highlight several key questions regarding the mechanism of translocation, and discuss possible mechanistic models in light of the recent crystal structures of the ribosome and its subunits. PMID- 11904174 TI - Ribosomal RNA pseudouridines and pseudouridine synthases. AB - Pseudouridines are found in virtually all ribosomal RNAs but their function is unknown. There are four to eight times more pseudouridines in eukaryotes than in eubacteria. Mapping 19 Haloarcula marismortui pseudouridines on the three dimensional 50S subunit does not show clustering. In bacteria, specific enzymes choose the site of pseudouridine formation. In eukaryotes, and probably also in archaea, selection and modification is done by a guide RNA-protein complex. No unique specific role for ribosomal pseudouridines has been identified. We propose that pseudouridine's function is as a molecular glue to stabilize required RNA conformations that would otherwise be too flexible. PMID- 11904176 TI - A tripeptide discriminator for stop codon recognition. AB - Only recently has it been established that a tripeptide in the bacterial release factors (RFs), RF1 and RF2, is responsible for the stop codon recognition. This functional mimic of the anticodon of tRNA is referred to as a tripeptide 'anticodon' or a tripeptide discriminator. Here we review the experimental background and process leading to this discovery, and strengthen functional evidence for the tripeptide determinant for deciphering stop codons in mRNAs in prokaryotes. PMID- 11904175 TI - Elongation factor-2 kinase and its newly discovered relatives. AB - Phosphorylation of elongation factor-2 (eEF-2) by the highly specific eEF-2 kinase results in eEF-2 inactivation and, therefore, may regulate the global rate of protein synthesis in animal cells. Cloning and sequencing of eEF-2 kinase led to the discovery of a new family of protein kinases, named alpha-kinases, whose catalytic domains display no sequence homology to conventional eukaryotic protein kinases. Several mammalian alpha-kinases have recently been cloned. Two of these alpha-kinases, named channel-kinases 1 and 2 (ChaK1 and ChaK2) represent a new type of signaling molecules that are protein kinases fused to ion channels. PMID- 11904177 TI - Cysteinyl-tRNA formation and prolyl-tRNA synthetase. AB - Aminoacyl-tRNA (AA-tRNA) formation is a key step in protein biosynthesis. This reaction is catalyzed with remarkable accuracy by the AA-tRNA synthetases, a family of 20 evolutionarily conserved enzymes. The lack of cysteinyl-tRNA (Cys tRNA) synthetase in some archaea gave rise to the discovery of the archaeal prolyl-tRNA (Pro-tRNA) synthetase, an enzyme capable of synthesizing Pro-tRNA and Cys-tRNA. Here we review our current knowledge of this fascinating process. PMID- 11904178 TI - The minimal tRNA: unique structure of Ascaris suum mitochondrial tRNA(Ser)(UCU) having a short T arm and lacking the entire D arm. AB - The tertiary structure of Ascaris suum mitochondrial tRNA(Ser)(UCU) was examined by nuclear magnetic resonance analysis using its transcript, since tRNA(Ser)(UCU), lacking the D arm and possessing a truncated T arm, is the shortest of all the known tRNAs. Most basepairs in the proposed secondary structure of tRNA(Ser)(UCU) were shown to exist, but the connector region comprising the truncated D loop and the extra loop was flexible. This flexibility, would enable adjustment of the mutual distance between the 3' terminus and the anticodon consistent with that of usual tRNAs. Thus, tRNA(Ser)UCU appears to function in a similar way to that of usual tRNAs in the ribosome. PMID- 11904179 TI - Decreased requirement for 4.5S RNA in 16S and 23S rRNA mutants of Escherichia coli. AB - 4.5S RNA is the bacterial homolog of the mammalian signal recognition particle (SRP) RNA that targets ribosome-bound nascent peptides to the endoplasmic reticulum. To explore the interaction of bacterial SRP with the ribosome, we have isolated rRNA suppressor mutations in Escherichia coli that decrease the requirement for 4.5S RNA. Mutations at C732 in 16S rRNA and at A1668 and G1423 in 23S rRNA altered the cellular responses to decreases in both Ffh (the bacterial homolog of SRP54) and 4.5S RNA levels, while the C1066U mutation in 16S rRNA and G424A mutation in 23S rRNA affected the requirement for 4.5S RNA only. These data are consistent with a dual role for 4.5S RNA, one involving co-translational protein secretion by a 4.5S-Ffh complex, the other involving free 4.5S RNA. PMID- 11904180 TI - Malignant transformation by the eukaryotic translation initiation factor 3 subunit p48 (eIF3e). AB - Several components of translation, e.g. eIF4E and PKR, are implicated in cancer. The e-subunit (p48) of mammalian initiation factor 3 is encoded by the Int6 gene, a common site for integration of the mouse mammary tumor virus genome, leading to the production of a truncated eukaryotic initiation factor-3e (eIF3e). Stable expression of a truncated eIF3e in NIH 3T3 cells causes malignant transformation by four criteria: foci formation; anchorage independent growth; accelerated growth; and lack of contact inhibition. Stable expression of full-length eIF3e does not cause transformation. The truncated eIF3e also inhibits the onset of apoptosis caused by serum starvation. PMID- 11904181 TI - How does tmRNA move through the ribosome? AB - To test the structure of tmRNA in solution, cross-linking experiments were performed which showed two sets of cross-links in two different domains of tmRNA. Site-directed mutagenesis was used to search for tmRNA nucleotide bases that might form a functional analogue of a codon-anticodon duplex to be recognized by the ribosomal A-site. We demonstrate that nucleotide residues U85 and A86 from tmRNA are significant for tmRNA function and propose that they are involved in formation of a tmRNA element playing a central role in A-site recognition. These data are discussed in the frame of a hypothetical model that suggests a general scheme for the interaction of tmRNA with the ribosome and explains how it moves through the ribosome. PMID- 11904182 TI - Transit of tRNA through the Escherichia coli ribosome: cross-linking of the 3' end of tRNA to ribosomal proteins at the P and E sites. AB - Photoreactive derivatives of yeast tRNA(Phe) containing 2-azidoadenosine at their 3' termini were used to trace the movement of tRNA across the 50S subunit during its transit from the P site to the E site of the 70S ribosome. When bound to the P site of poly(U)-programmed ribosomes, deacylated tRNA(Phe), Phe-tRNA(Phe) and N acetyl-Phe-tRNA(Phe) probes labeled protein L27 and two main sites within domain V of the 23S RNA. In contrast, deacylated tRNA(Phe) bound to the E site in the presence of poly(U) labeled protein L33 and a single site within domain V of the 23S rRNA. In the absence of poly(U), the deacylated tRNA(Phe) probe also labeled protein L1. Cross-linking experiments with vacant 70S ribosomes revealed that deacylated tRNA enters the P site through the E site, progressively labeling proteins L1, L33 and, finally, L27. In the course of this process, tRNA passes through the intermediate P/E binding state. These findings suggest that the transit of tRNA from the P site to the E site involves the same interactions, but in reverse order. Moreover, our results indicate that the final release of deacylated tRNA from the ribosome is mediated by the F site, for which protein L1 serves as a marker. The results also show that the precise placement of the acceptor end of tRNA on the 50S subunit at the P and E sites is influenced in subtle ways both by the presence of aminoacyl or peptidyl moieties and, more surprisingly, by the environment of the anticodon on the 30S subunit. PMID- 11904183 TI - Elongation factor G with effector loop from elongation factor Tu is inactive in translocation. AB - Elongation factors Tu and G (EF-Tu and EF-G) alternately interact with the ribosome during the elongation phase of protein biosynthesis. The function of both factors depends on GTP binding, and the factors are ascribed to a superfamily of G-proteins. All G-proteins contain the effector loop, a structural element that is important for the protein's interaction with its target molecule. In this study the effector loop of EF-G was replaced by the loop taken from EF Tu. The EF-G with EF-Tu loop has markedly decreased GTPase activity and did not catalyze translocation. We conclude that these loops are not functionally interchangeable since the factors interact with different states of the ribosome. PMID- 11904184 TI - Construction of the 'minimal' SRP that interacts with the translating ribosome but not with specific membrane receptors in Escherichia coli. AB - Escherichia coli signal recognition particle (SRP) consists of 4.5S RNA and Ffh protein. In contrast to eukaryotes, it remains unclear whether translation arrest takes place in prokaryotic cells. To study this problem we constructed a fusion of the M domain of Ffh protein with a cleavable affinity tag. This mutant Ffh, in a complex with 4.5S RNA, can bind signal peptide at the translating ribosome but is unable to bind the membrane. This SRP-ribosome complex should accumulate in the cell if translation is arrested. To test this, the complex was purified from the cells by ultracentrifugation and affinity chromatography. The composition of the complex was analyzed and found to consist of ribosomal RNAs and proteins, the Ffh M domain and 4.5S RNA. The accumulation of this complex in the cell in significant amounts indicated that SRP-mediated translation arrest did occur in bacterial cells. PMID- 11904185 TI - The role of SmpB protein in trans-translation. AB - The function of SmpB protein in the trans-translation system was evaluated using the well-defined cell-free translation system consisting of purified ribosome, alanyl-tRNA synthetase and elongation factors. The analysis showed that SmpB protein enhances alanine-accepting activity of tmRNA and that SmpB protein and tmRNA are sufficient to complete the trans-translation process in the presence of translational components. Moreover, SmpB is indispensable in the addition of tag peptide onto ribosomes by tmRNA. In particular, the A-site binding of tmRNA is inhibited in the absence of SmpB. PMID- 11904186 TI - Functional evidence for D- and T-loop interactions in tmRNA. AB - During bacterial protein synthesis, stalled ribosomes can be rescued by tmRNA, a molecule with both tRNA and mRNA features. The tRNA region of tmRNA has sequence similarity with tRNA(Ala) and also has a clover-leaf structure folded similarly as in canonical tRNAs. Here we propose the L-shape of tmRNA to be stabilized by two tertiary interactions between its D- and T-loop on the basis of phylogenetic and experimental evidence. Mutational analysis clearly demonstrates a tertiary interaction between G(13) and U(342). Strikingly, this in evolution conserved interaction is not primarily important for tmRNA alanylation and for binding to elongation factor Tu, but especially for a proper functioning of SmpB. PMID- 11904187 TI - Tandem termination signals: myth or reality? AB - In two Escherichia coli genomes, laboratory strain K-12 and pathological strain O157:H7, tandem termination codons as a group are slightly over-represented as termination signals. Individually however, they span the range of representations, over, as expected, or under, in one or both of the strains. In vivo, tandem termination codons do not make more efficient signals. The second codon can act as a backstop where readthrough of the first has occurred, but not at the expected efficiency. UGAUGA remains an enigma, highly over-represented, but with the second UGA a relatively inefficient back up stop codon. PMID- 11904188 TI - RNA aptamers directed against release factor 1 from Thermus thermophilus. AB - An in vitro selection/amplification (SELEX) was used to generate RNA aptamers that specifically bind Thermus thermophilus release factor 1 (RF1). From 31 isolated clones, two groups of aptamers with invariable sequences 5'-ACCU-3' and 5'-GAAAGC-3' were isolated. Chemical and enzymatic probing of the structure indicate that in both groups the invariable sequences are located in single stranded regions of hairpin structures. Complex formations between RF1 and aptamers of both groups were identified by electrophoretic shift assay and chemical footprinting. Deletion of the invariable sequences did not effect the secondary structure of the aptamers but abolished their binding to RF1. RNA motifs matching the invariable sequences of the aptamers are present as consensus sequences in the peptidyl transferase center of 23S rRNAs. T. thermophilus RF1 recognizes UAG stop codons in an Escherichia coli in vitro translation system. Aptamers from both groups inhibited this RF1 activity. PMID- 11904189 TI - Positioning of the mRNA stop signal with respect to polypeptide chain release factors and ribosomal proteins in 80S ribosomes. AB - To study positioning of the mRNA stop signal with respect to polypeptide chain release factors (RFs) and ribosomal components within human 80S ribosomes, photoreactive mRNA analogs were applied. Derivatives of the UUCUAAA heptaribonucleotide containing the UUC codon for Phe and the stop signal UAAA, which bore a perfluoroaryl azido group at either the fourth nucleotide or the 3' terminal phosphate, were synthesized. The UUC codon was directed to the ribosomal P site by the cognate tRNA(Phe), targeting the UAA stop codon to the A site. Mild UV irradiation of the ternary complexes consisting of the 80S ribosome, the mRNA analog and tRNA resulted in tRNA-dependent crosslinking of the mRNA analogs to the 40S ribosomal proteins and the 18S rRNA. mRNA analogs with the photoreactive group at the fourth uridine (the first base of the stop codon) crosslinked mainly to protein S15 (and much less to S2). For the 3'-modified mRNA analog, the major crosslinking target was protein S2, while protein S15 was much less crosslinked. Crosslinking of eukaryotic (e) RF1 was entirely dependent on the presence of a stop signal in the mRNA analog. eRF3 in the presence of eRF1 did not crosslink, but decreased the yield of eRF1 crosslinking. We conclude that (i) proteins S15 and S2 of the 40S ribosomal subunit are located near the A site-bound codon; (ii) eRF1 can induce spatial rearrangement of the 80S ribosome leading to movement of protein L4 of the 60S ribosomal subunit closer to the codon located at the A site; (iii) within the 80S ribosome, eRF3 in the presence of eRF1 does not contact the stop codon at the A site and is probably located mostly (if not entirely) on the 60S subunit. PMID- 11904190 TI - A bilayer cell-free protein synthesis system for high-throughput screening of gene products. AB - A high-throughput cell-free protein synthesis method has been described. The methodology is based on a bilayer diffusion system that enables the continuous supply of substrates, together with the continuous removal of small byproducts, through a phase between the translation mixture and substrate mixture. With the use of a multititer plate the system was functional for a prolonged time, and as a consequence yielded more than 10 times that of the similar batch-mode reaction. Combining this method with a wheat germ cell-free translation system developed by us, the system could produce a large amount of protein sufficient for carrying out functional analyses. This novel bilayer-based cell-free protein synthesis system with its simplicity, minimum time and low cost may be useful practical methodology in the post-genome era. PMID- 11904191 TI - Ribosome display for selection of active dihydrofolate reductase mutants using immobilized methotrexate on agarose beads. AB - Ribosome display was applied to the selection of an enzyme. As a model, we selected and amplified the dihydrofolate reductase (DHFR) gene by ribosome display utilizing a wheat germ cell-free protein synthesis system based on binding affinity to its substrate analog, methotrexate, immobilized on agarose beads. After three rounds of selection, the DHFR gene could be effectively selected and preferentially amplified from a small proportion in a mixture also containing competitive genes. Active enzymes were expressed and amplified and by sequence analysis, four mutants of DHFR were identified. These mutants showed as much activity as the wild-type enzyme. PMID- 11904193 TI - Spontaneous disc space infections in adults. AB - BACKGROUND: Spontaneous discitis typically affects children; much less is known about the disease in adults. We examined the clinical characteristics and the role of surgery in spontaneous infectious discitis in adults. METHODS: Twenty nine consecutive adult patients (16 men, 13 women) with spontaneous infectious discitis were treated by a single surgeon (MJE) over a 5-year period. These patients were compared to 19 consecutive patients with postoperative discitis over the same time period. Mean follow-up was 2.4 years. RESULTS: The average age of patients with spontaneous discitis was 69.0 years. Eleven of these patients (38%) were diabetic and 9 (31%) had a known concurrent infection. Infections occurred at cervical (10%), thoracic (34%), and lumbar interspaces (59%), and at multiple disc spaces in eight cases (28%). Ten patients (34%) had an elevated serum leukocyte count and 21 patients (72%) had an elevated erythrocyte sedimentation rate. While Gram-positive infection was most common, a broad spectrum of microbes was isolated. All patients were treated with i.v. antibiotics for a mean duration of 6 weeks. Four patients required surgical fusion; the rest were treated with external immobilization. At follow-up, 21 patients (72%) were symptom-free, 3 patients (10%) had ongoing disease, and 5 patients (17%) had died of unrelated causes. Compared to patients with postoperative discitis treated over the same time period, spontaneous discitis in adults affected older patients and required a broader spectrum of antibiotic coverage. Outcomes were similar between the two groups. CONCLUSIONS: While spontaneous discitis has predominantly been reported in children, the disease also affects older adults. Spontaneous discitis in adults is associated with advanced age, diabetes mellitus, and systemic infection. Elevated serum leukocyte count lacks diagnostic sensitivity. A high cure rate is achieved with antibiotics and external immobilization. Surgical treatment is rarely required. PMID- 11904198 TI - Selective posterior rhizotomy for lower extremity spasticity: how much and which of the posterior rootlets should be cut? AB - BACKGROUND: It is well known that selective posterior rhizotomy is effective for relieving spasticity associated with cerebral palsy. However, there is significant variation between surgeons in terms of how much and which of the posterior rootlets should be cut for the improvement of ambulatory function without causing adverse effects. METHODS: The study population was composed of 200 CP patients who underwent SPR more than 1 year before this study. The children were divided into 4 groups (Group A had their L1-S2 roots cut, Group B had the L2-S2 roots cut, Group C had the L2-S1 roots cut, and Group D had the L2 S1 roots and the unilateral S2 root cut). We assessed lower limb spasticity, passive range of motion, ambulatory function, and gait pattern in each group. RESULTS: Inclusion of L1 and S2 in the lesioning process of SPR was more effective at relieving spasticity in terms of hip adduction and ankle dorsiflexion respectively and improving ambulatory function (p < 0.01). Although lesioning of S2 carried a greater risk of urinary dysfunction, resection of less than 50% of S2 significantly improved ambulatory function without urinary complications (p < 0.01). Unilateral lesioning of S2 was an alternative option in selected cases with different amounts of spasticity in the ankles for the same purpose. CONCLUSIONS: We propose that L1 and S2 roots should be included in the lesioning process of SPR for effective improvement of gross motor function, but that resection of these roots should be less than 50% to prevent complications. PMID- 11904199 TI - Insertion of vagal nerve stimulator using local and regional anesthesia. AB - BACKGROUND: Vagal nerve stimulation (VNS) is a valuable therapy for patients with intractable epilepsy. Placement of a vagal nerve stimulator typically requires general anesthesia, which frequently interrupts anticonvulsant therapy. Insertion of the stimulator using regional/local anesthesia may offer the advantages of continuity of anticonvulsant therapy and implantation in the outpatient setting. METHODS: We retrospectively compared the first 10 consecutive patients undergoing VNS implantation under general anesthesia with the first 12 consecutive patients undergoing VNS implantation under regional/local anesthesia. Patients for the regional/local anesthesia were selected on the basis of their ability to cooperate and follow commands. Regional anesthesia for implantation of the VNS leads was achieved by performing superficial and deep cervical plexus blocks. A local anesthetic field block of a small area of the posterior chest provided anesthesia for insertion of the generator. RESULTS: All of the patients undergoing regional/local anesthesia completed the procedure without difficulty and on an outpatient basis. None complained of discomfort, sedation, nausea, or vomiting and none had seizures in the perioperative period. These results contrasted with the group that underwent general anesthesia (n = 10), who had an 80% incidence of nausea and vomiting and a 30% incidence of postoperative seizures. CONCLUSION: VNS implantation under regional/local anesthesia is proficiently performed as an outpatient procedure with minimal postoperative side effects. PMID- 11904200 TI - Giant prolactinomas presenting as skull base tumors. AB - BACKGROUND: Prolactinomas invading the skull base are rare, and could easily be confused with skull base tumors of nonpituitary origin. CASE DESCRIPTION: We report a series of 4 cases of giant prolactinomas invading the skull base and presenting with atypical symptoms. Case 1 presented with a short history of headache and nasal obstruction. Case 2 presented with progressive hypoacusia, dizziness, and ophthalmoplegia. In Case 3, the patient developed rapid progressive visual failure and psychiatric symptoms. Case 4 presented with a 1 year history of headache and retrorbital pain. The diagnosis of prolactinoma was made on the basis of tumor immunohistochemistry and/or high plasma prolactin levels (range from 650-6,500 ng/mL). Medical treatment with the dopamine agonist cabergoline was given; it was effective in normalizing prolactin levels and inducing tumor shrinkage. CONCLUSION: Prolactin levels should be measured in all large skull base tumors involving the pituitary region before any surgery or inappropriate radiotherapy is performed. PMID- 11904203 TI - Hypervascular vestibular schwannomas. AB - BACKGROUND: Vestibular schwannoma (VS) is usually hypovascular and can be resected totally without major morbidity. Resection of the more uncommon hypervascular VS is complicated by excessive tumor bleeding. We have attempted to clarify the clinical characteristics and management of hypervascular VS. METHODS: Surgical reports and videos of 78 patients with unilateral VS (5 hypervascular, 73 nonhypervascular) were retrospectively reviewed and clinical characteristics, radiological findings, and case management were compared. RESULTS: Hypervascular VS presented at a younger age than nonhypervascular VS (29 +/- 12 vs. 52 +/- 16 years old) (p < 0.01). Magnetic resonance imaging (MRI) showed that hypervascular VS was solid, without tumor cyst, and significantly larger than nonhypervascular VS (p < 0.05). The surface of hypervascular VS consistently showed multiple flow voids representing large draining veins. The characteristic angiographical findings of hypervascular VS were extensive tumor vessels, tumor stains, and early filling of draining veins; vertebrobasilar arteries supplied hypervascular VS. A multi-stage surgical approach was used since torrential tumor bleeding in the first surgery interfered with resection, resulting in partial tumor removal. Angiography before the second surgery showed much reduced tumor vascularity, bleeding was much reduced, and tumor was resected with less difficulty. In this approach, all 5 hypervascular VS were resected totally (1 case) or near-totally (4 cases) without major morbidity. CONCLUSIONS: Hypervascular VS, a solid and large tumor, presents at an earlier age. Although angiography provides characteristic findings, MRI can confirm the diagnosis of a hypervascular VS by showing multiple flow-voids on the tumor surface. Since partial tumor removal (first surgery) extensively reduces tumor vascularity and intraoperative tumor bleeding considerably, hypervascular VS should be managed by a multi-staged surgical approach. PMID- 11904205 TI - A study of primary central nervous system lymphoma in northern India. AB - BACKGROUND: Primary central nervous system lymphoma (PCNSL) is rare and accounts for 1 to 2% of all lymphomas. There are conflicting reports about the rise in incidence of PCNSL cases in the last two decades; this has largely been attributed to an increase in incidence of AIDS and other immunosuppressive states in some studies. This study was undertaken to view the trend of PCNSL at the Postgraduate Institute of Medical Education and Research, Chandigarh, which is a referral hospital in northern India. METHODS: The PCNSL cases from our surgical material of 15 years (1985-1999) were reviewed. Slides were examined independently by three histopathologists. Immunophenotyping was done on paraffin embedded tissue using indirect immunoperoxidase technique. RESULTS: Out of a total of 3,325 intracranial tumors diagnosed during this period (1985-1999), there were 40 cases (1.2%) of PCNSL; gliomas accounted for 1,531 cases (46.04%). The age ranged from 24-75 years with the sex ratio (M:F) being 2:1. HIV serology, available in 14 cases, was negative in all. The parietal lobe was the most common site of involvement. Diffuse large cell lymphoma was the most common morphological type. Immunohistochemistry could be done in 31 cases; 28 cases were found to be B-cell type whereas 3 cases were T-cell type. No statistically significant increase was seen on comparing the number of cases at 5-year intervals. CONCLUSION: This study revealed that there has been no significant increase in PCNSL cases over the last 15 years. PMID- 11904207 TI - Traumatic encephalocele related to orbital roof fractures: report of six cases and literature review. AB - BACKGROUND: Orbital roof fractures after blunt injury are rare. Traumatic encephaloceles in the orbital cavity are even rarer, with only 15 cases published to date. METHODS: The clinical, radiological, and surgical findings of 6 cases of traumatic encephalocele treated at our institution from June 1998 to January 2000 are presented. They are also compared with previously published series. RESULTS: In contrast to other published cases, 5 out of 6 patients in our series were adults. The most common cause of trauma was road traffic accident. Ecchymosis and preoperative exophthalmos/proptosis were frequent. In all of our patients a coronal CT scan (3 mm increments with bone windows) was obtained. It demonstrated the extension of the orbital roof fractures and a possible encephalocele in 4 cases. Associated frontal brain contusions were seen in 5 cases. An MRI was performed in 3 patients (and only in 2 previously published cases); it showed the extension of the brain herniation into the orbital cavity. Surgical treatment via a fronto-basal approach with evacuation of the contused herniated brain tissue and orbital roof reconstruction was performed. The outcome at 6 months was good recovery in five patients with one patient still in a persistent vegetative state. Postoperatively the ocular disturbances improved in 5 cases. A review of the other published cases confirmed recovery of normal ocular function in the vast majority of the cases. CONCLUSIONS: Whenever orbital roof fractures associated with frontal contusions are identified in an acute brain injured patient, an orbital encephalocele should be suspected. In our opinion MRI is the investigation of choice in such patients. If the encephalocele is confirmed, a surgical approach via the subfrontal route is indicated with resection of herniated contused brain tissue, dural closure, and orbital roof reconstruction. Good results in regard to the orbital symptoms (mainly exophthalmos) can be expected. PMID- 11904208 TI - Vascular tunnel creation to improve the efficacy of decompressive craniotomy in post-traumatic cerebral edema and ischemic stroke. AB - BACKGROUND: Decompressive craniectomy with durotomy has been well described but remains a controversial treatment for traumatic or ischemic brain swelling. Although the technique can reduce intracranial pressure it frequently results in infarction of the brain tissue which extends through the durotomy because of compression and subsequent complications associated with decompressive craniectomy. METHODS: All patients treated with surgical decompression were comatose with Grade 3 or 4 Glasgow Coma Scores. Since 1998 we have changed our technique for decompressive craniectomy by creating vascular channels around the major vessels crossing the durotomy margin. Outcomes in 21 patients treated with this technique are compared to 20 patients treated with a conventional decompression and durotomy between 1997 through 1999. RESULTS: Clinical outcome was substantially better in the group treated with a vascular tunnel. CONCLUSIONS: Our results suggest that placing vascular tunnels at the margins of a durotomy when performing a decompression may reduce vascular congestion and the subsequent ischemia in brain tissue which herniates through the durotomy, leading to better clinical outcomes. PMID- 11904210 TI - Complete penetration of the optic chiasm by an unruptured aneurysm of the ophthalmic segment: case report. AB - BACKGROUND: It is well known that aneurysms of the ophthalmic segment sometimes elevate the optic nerve or chiasm, and in case of large or giant aneurysms, the optic apparatus can be dramatically thinned. Nonetheless, they rarely penetrate the optic pathway completely. To our knowledge, no previous reports have dealt with the complete penetration of the optic chiasm by unruptured aneurysms of the ophthalmic segment. CASE DESCRIPTION: A 70-year-old woman presented with visual dysfunction in her left eye that she had experienced for several months. Her left visual acuity had rapidly deteriorated to the level of finger counting and visual field testing demonstrated nasal hemianopsia in the left eye and upper temporal quadrant hemianopsia in the right eye. Left internal carotid angiograms and three dimensional digital subtraction angiograms showed an aneurysm of the ophthalmic segment projecting superomedially. Intraoperative findings revealed complete penetration of the optic chiasm by the fundus of the aneurysm. The optic pathway adjacent to the dome had become remarkably thin and dark yellow. After clipping was completed, the fundus of the aneurysm was punctured to decompress the optic chiasm. Postoperatively, patient's visual acuity in the left eye gradually recovered, but the visual field deficit persisted after the operation. CONCLUSION: This rare case demonstrates the potentially aggressive behavior of unruptured aneurysms of the ophthalmic segment. Patients with unruptured aneurysms of the ophthalmic segment who present with visual symptoms should be treated with surgical clipping to decompress the optic pathway as soon as possible. PMID- 11904212 TI - Thoughts on strategic retreats and group governance. PMID- 11904213 TI - How to educate neurosurgeons from other countries: one group's experience. PMID- 11904216 TI - The "When All Else Fails" syndrome. PMID- 11904214 TI - Plating for ACDF. PMID- 11904217 TI - The wild type bacterial Co(2+)/Co(2+)-phosphotriesterase shows a middle-range thermostability. AB - The phosphotriesterase (PTE) from Pseudomonas diminuta, a metalloenzyme that catalyses the hydrolysis of organophosphorus pesticides and nerve agents, has been described as a remarkably heat-stable protein [Grimsley et al., Biochemistry 36 (1997), 14366-14374]. Because substitution of the naturally occurring zinc ions by cobalt ions was found to enhance the enzyme catalytic activity, we investigated the thermal stability of the Co(2+)/Co(2+)-PTE. This study, carried out using capillary electrophoresis under optimised conditions in the pH range 9 10 compatible with optimal enzyme activity, provided evidence for irreversible denaturation according to the Lumry-Eyring model. A temperature-induced conformational transition (T(m) approximately equal to 58 degrees C) and an early growing of aggregates were observed. Comparison of UV spectra with heat-induced inactivation data clearly demonstrated that the PTE state populated above T(m) was neither native nor active. Differential scanning calorimetry showed only an exothermic trace due to aggregation of the denatured protein at T=76 degrees C. Accordingly, the temperature-induced denaturation process of the PTE could be described by a consecutive reaction model, including formation of an intermediate with enhanced activity at T approximately equal to 45 degrees C and an inactive unfolded state populated at T approximately equal to 58 degrees C, which leads to denatured aggregates. Thus, the wild type Co(2+)/Co(2+)-PTE displays a middle range thermostability. Hence, for decontamination purposes under extreme Earth temperatures, wild type and engineered mutants of PTE substituted with other metal cations should be evaluated. PMID- 11904218 TI - Recombinant lysine:N(6)-hydroxylase: effect of cysteine-->alanine replacements on structural integrity and catalytic competence. AB - Recombinant lysine:N(6)-hydroxylase, rIucD, catalyzes the hydroxylation of L lysine to its N(6)-hydroxy derivative, with NADPH and FAD serving as cofactors in the reaction. The five cysteine residues present in rIucD can be replaced, individually or in combination, with alanine without effecting a major change in the thermal stability, the affinity for L-lysine and FAD, as well as the k(cat) for mono-oxygenase activity of the protein. However, when the susceptibility to modification by either 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) or 2,6 dichlorophenol indophenol (DPIP) serves as the criterion for monitoring conformational change(s) in rIucD and its muteins, Cys146-->Ala and Cys166-->Ala substitutions are found to induce an enhancement in the reactivity of one of the protein's remaining cysteine residues with concomitant diminution of mono oxygenase function. In addition, the systematic study of cysteine-->alanine replacement has led to the identification of rIucD's Cys166 as the exposed residue which is detectable during the reaction of the protein with DTNB but not with iodoacetate. Substitution of Cys51 of rIucD with alanine results in an increase in mono-oxygenase activity (approx. 2-fold). Such replacement, unlike those of other cysteine residues, also enables the covalent DPIP conjugate of the protein to accommodate FAD in its catalytic function. A possible role of rIucD's Cys51 in the modulation of its mono-oxygenase function is discussed. PMID- 11904219 TI - Inhibition of nuclear import by backbone cyclic peptidomimetics derived from the HIV-1 MA NLS sequence. AB - In the present work we have constructed a series of backbone cyclic peptides, which differed in the amino acid residues located at the C-terminal position of the previously described BCvir peptide (A. Friedler, N. Zakai, O. Karni, Y.C. Broder, L. Baraz, M. Kotler, A. Loyter, C. Gilon, Biochemistry 37 (1998)). BCvir is a cyclic peptide, derived from the nuclear localization signal (NLS) of the human immunodeficiency virus type 1 matrix protein. The majority of the cyclic peptides described here inhibited nuclear import in vitro. The most potent inhibitors were those bearing bulky hydrophobic amino acids such as Leu, Phe or Nal (naphthyl Ala) at the C-terminus. On the other hand, peptides bearing polar amino acid residues such as Asn, Cys or a reduced amide bond were not inhibitory. The present studies demonstrate the importance of a bulky hydrophobic C-terminal side chain and an exocyclic amide bond preceding it, to the inhibitory activity of the NLS-derived BC peptides. Being only inhibitory, these BC peptides resemble classic receptor antagonists. PMID- 11904220 TI - The inhibitory form of NifL from Klebsiella pneumoniae exhibits ATP hydrolyzing activity only when synthesized under nitrogen sufficiency. AB - The inhibitory function of Klebsiella pneumoniae NifL on NifA transcriptional activity in vitro is stimulated by ATP and ADP when NifL is synthesized under nitrogen sufficiency (NifL(NH4)). Further characterizations showed that NifL(NH4) binds and hydrolyzes ATP (2500 mU/mg). Analyzing fusions between MalE and different portions of NifL, we localized both the ATP binding site and ATP hydrolysis activity to the N-terminal domain of NifL. In contrast, NifL synthesized under nitrogen limitation is not affected by adenine nucleotides and exhibits no ATP hydrolyzing activity. These major differences indicate that the stimulation of the inhibitory function of NifL and the ability to hydrolyze ATP depend on a specific NifL conformation induced by ammonium. We hypothesize that the presence of ammonium alters the conformation of NifL, enabling it to use the energy of ATP hydrolysis to increase the efficiency of NifL-NifA complex formation. PMID- 11904221 TI - High expression in adult horse of PLRP2 displaying a low phospholipase activity. AB - The physiological role of the two lipase-related proteins, PLRP1 and PLRP2, still remains obscure although some propositions have been made concerning PLRP2. In this paper, we report the presence of high amounts of PLRP2 in adult horse pancreas whereas no PLRP1 could be detected. As well, a non-parallel expression of PLRP2 and PLRP1 is observed in adult cat and dog, since no PLRP2 could be detected in these two species. In adult ox, neither PLRP2 nor PLRP1 could be found. These findings are in favor of a different regulation of the expression of the genes encoding pancreatic lipase and the related proteins according to the species. The cDNA encoding horse PLRP2 has been cloned and the protein expressed in insect cells. Both native and recombinant PLRP2 display the same catalytic properties. They possess a moderate lipase activity, inhibited by bile salts and not restored by colipase. Interestingly, they differ from PLRP2 from other species by their very low phospholipase activity indicating that PLRP2 could not be considered as a general phospholipase as previously postulated. This work highlights the variability of the properties of PLRP2 and rises the question of the physiological function of this protein in adult according to the species. PMID- 11904222 TI - Polydispersity of Bacillus thuringiensis Cry1 toxins in solution and its effect on receptor binding kinetics. AB - Dynamic light scattering and surface plasmon resonance techniques were used to investigate the influence of ionic strength, buffer composition and pH on the multimerization of trypsin-activated Cry1Ac and Cry1C toxins over time and the subsequent effects of the different multimers on receptor binding models. In carbonate buffer at pH 10.5, Cry1Ac and Cry1C assumed a monomeric state. After 24 h, a complete conversion of monomeric toxin to a dimeric or trimeric form was observed only for Cry1Ac under low ionic strength condition. Cry1C and Cry1Ac in high ionic strength buffer remained monomeric. Substitution of CAPS pH 11 for carbonate buffer suppressed this Cry1Ac oligomerization effect. Once Cry1Ac toxin was in an aggregated form, increases in ionic strength failed to revert the aggregated toxin back to a monomeric form. Monomeric Cry1Ac bound to a purified 115 kDa aminopeptidase N receptor from Manduca sexta in a 2:1 molar ratio thus confirming the existence of two binding sites on this receptor. Binding rates of dimeric or higher aggregated Cry1Ac toxin forms were different from those generated using the monomeric form and could not be fitted to existing binding models. In summary, our results confirm that the M. sexta 115 kDa aminopeptidase N receptor possesses two Cry1Ac binding sites. They further suggest that although high pH and low salt conditions promote Cry1Ac aggregation, this observation cannot be applied universally to other members of the Cry family. PMID- 11904223 TI - The 3-D structure of microsomal glutathione transferase 1 at 6 A resolution as determined by electron crystallography of p22(1)2(1) crystals. AB - Pure solubilised microsomal glutathione transferase 1 (MGST1) forms well-ordered two-dimensional (2-D) crystals of two different symmetries, one orthorhombic (p22(1)2(1)) and one hexagonal (p6), both diffracting electrons to a resolution beyond 3 A. A three-dimensional (3-D) map has previously been calculated to 6 A resolution from the hexagonal crystal form. From orthorhombic crystals we have now calculated a 6 A 3-D reconstruction displaying three repeats of four rod-like densities. These are inclined relative to the normal of the membrane plane and consistent with arising from a left-handed four-helix bundle fold. The rendered volume clearly displays the same structural features as the map previously calculated from the p6 crystal type including similar lengths and substructure of the helices, but several distinguishing features do exist. The helices are more tilted in the map calculated from the orthorhombic crystals indicating conformational flexibility. Density present on the cytosolic side is consistent with the location of the active site. In addition, the current map displays the noted similarity to subunit I of cytochrome c oxidase. PMID- 11904224 TI - Folding and stability of the C-terminal half of apolipoprotein A-I examined with a Cys-specific fluorescence probe. AB - Apolipoprotein A-I (apoA-I) has important physiologic roles in reverse cholesterol transport, as a component of HDL; however, apoA-I also exists in lipid-poor or lipid-free forms that are key intermediates in HDL metabolism and acceptors of lipids from cells. The aim of this study was to examine the structure and stability of the central and C-terminal regions of lipid-free apoA I. To this end, five Cys mutants of proapoA-I were constructed and expressed in Escherichia coli: V119C, A124C, A154C, A190C, and A232C. These mutants were specifically labeled with 6-acryloyl-2-dimethylaminonaphthalene (acrylodan, AC) and were examined by CD spectroscopy and a variety of fluorescence methods. The results showed that the introduction of Cys residues and their covalent labeling with AC did not affect the overall structure and stability of apoA-I. However, AC fluorescence properties revealed that different segments of the central and C terminal half of apoA-I have distinct folding and stability properties. From fluorescence energy transfer data, average distances between the N-terminal region containing Trp residues and the various AC locations were obtained. The current results, together with previously published observations, led to the construction of a three-dimensional model for the folding of lipid-free apoA-I. PMID- 11904225 TI - Helicobacter pylori 3-deoxy-D-manno-octulosonate-8-phosphate (KDO-8-P) synthase is a zinc-metalloenzyme. AB - 3-Deoxy-D-manno-2-octulosonate-8-phosphate (KDO-8-P) synthase catalyzes the aldol type condensation of phosphoenolpyruvate and D-arabinose-5-phosphate (A-5-P) to produce KDO-8-P and inorganic phosphate. All KDO-8-P synthases, as exemplified by the enzyme from Escherichia coli, were believed not to require a metal cofactor for catalytic activity. However, recent studies have demonstrated that the KDO-8 P synthase from Aquifex aeolicus is a metalloenzyme. Moreover, sequence alignments and phylogenetic analysis of KDO-8-P synthase protein sequences strongly suggested that there is a whole subfamily of KDO-8-P synthases that are also metalloenzymes. One of these putative metalloenzymes is the ortholog from the human pathogen Helicobacter pylori. In order to test this model, we have cloned the kdsa gene encoding H. pylori KDO-8-P synthase, and overexpressed and purified the protein. This enzyme was found to bind one mol Zn/mol monomer, and the removal of this metal by treatment with 2,6-pyridine dicarboxylic acid abolished enzymatic activity. The Zn(2+) in the enzyme could be quantitatively replaced by Cd(2+), which increased the observed k(cat) by approximately 2-fold, and decreased the apparent K(m)(A-5-P) by approximately 6.5-fold. Furthermore, removal of the Zn(2+) from the enzyme did not greatly perturb its circular dichroism spectra. Thus, the divalent metal most likely serves as cofactor directly involved in catalysis. PMID- 11904226 TI - Changes at the interface of the N- and C-terminal parts of the heavy chain of myosin subfragment 1. AB - It has been previously shown that in the M-MgADP-P(i) state, where the myosin head adopts a pre-power stroke conformation, treatment of trypsin-split subfragment 1 of skeletal muscle myosin with 1-ethyl-3-(3 dimethylaminopropyl)carbodiimide (EDC) results in cross-linking of the C-terminal fragment of the heavy chain of S1 -- most probably its converter region -- to the N-terminal S1 heavy-chain fragment, generating a product of 44 kDa [Biochim. Biophys. Acta 1481 (2000) 55]. The results described here show that this product is neither generated in the absence of nucleotide nor in the presence of MgADP or MgPP(i). The 44 kDa cross-linking product can be formed when S1 treated with EDC is complexed with MgADP-AlF(4) or MgADP-V(i) (MgADP-P(i) analogs) and with MgADP BeF(x), MgATP gamma S or MgAMPPNP (MgATP analogs). The results suggest structural differences between MgATP- or MgADP-P(i)-bound S1, and MgADP-bound or nucleotide free S1, in spatially close regions of their N- and C-terminal heavy-chain fragments. PMID- 11904228 TI - Size as a parameter for solvent effects on Candida antarctica lipase B enantioselectivity. AB - Changes in solvent type were shown to yield significant improvement of enzyme enantioselectivity. The resolution of 3-methyl-2-butanol catalyzed by Candida antarctica lipase B, CALB, was studied in eight liquid organic solvents and supercritical carbon dioxide, SCCO(2). Studies of the temperature dependence of the enantiomeric ratio allowed determination of the enthalpic (Delta(R-S)Delta H(++)) as well as the entropic (Delta(R-S)Delta S(++)) contribution to the overall enantioselectivity (Delta(R-S)Delta G(++)= -RTlnE). A correlation of the enantiomeric ratio, E, to the van der Waals volume of the solvent molecules was observed and suggested as one of the parameters that govern solvent effects on enzyme catalysis. An enthalpy-entropy compensation relationship was indicated between the studied liquid solvents. The enzymatic mechanism must be of a somewhat different nature in SCCO(2), as this reaction in this medium did not follow the enthalpy-entropy compensation relation. PMID- 11904227 TI - Substrate activation in acetylcholinesterase induced by low pH or mutation in the pi-cation subsite. AB - Substrate inhibition is considered a defining property of acetylcholinesterase (AChE), whereas substrate activation is characteristic of butyrylcholinesterase (BuChE). To understand the mechanism of substrate inhibition, the pH dependence of acetylthiocholine hydrolysis by AChE was studied between pH 5 and 8. Wild-type human AChE and its mutants Y337G and Y337W, as well as wild-type Bungarus fasciatus AChE and its mutants Y333G, Y333A and Y333W were studied. The pH profile results were unexpected. Instead of substrate inhibition, wild-type AChE and all mutants showed substrate activation at low pH. At high pH, there was substrate inhibition for wild-type AChE and for the mutant with tryptophan in the pi-cation subsite, but substrate activation for mutants containing small residues, glycine or alanine. This is particularly apparent in the B. fasciatus AChE. Thus a single amino acid substitution in the pi-cation site, from the aromatic tyrosine of B. fasciatus AChE to the alanine of BuChE, caused AChE to behave like BuChE. Excess substrate binds to the peripheral anionic site (PAS) of AChE. The finding that AChE is activated by excess substrate supports the idea that binding of a second substrate molecule to the PAS induces a conformational change that reorganizes the active site. PMID- 11904230 TI - The effect of anions on azide binding to myoglobin: an unusual functional modulation. AB - The effect of increasing concentrations of several anions on the azide (N(-)(3)) binding properties of sperm whale and horse ferric myoglobin has been studied. Surprisingly, a number of anions may act as heterotropic effectors, decreasing the affinity of myoglobins for N(-)(3), in the following order: ClO(-)(4)=I( )>Br(-)>Cl(-) and SO(2-)(4), which mirrors the increase in their charge density. The largest effects were measured using ClO(-)(4) and I(-), which produce a 4 fold and 8-fold reduction of the N(-)(3) binding affinity in horse and sperm whale myoglobins, respectively. A dissociation equilibrium constant (K(d)) ranging from 150 to 250 mM was estimated for ClO(-)(4) and I(-) binding to myoglobins. In order to analyse the molecular mechanism producing the reduction of the N(-)(3) binding affinity to ferric myoglobin, the potential anionic binding sites within ferric myoglobin were investigated by a molecular modelling study using the program Grid. Analysis of the theoretical results suggests two particularly favourable binding sites: the first, next to the distal side of the haem, whose occupancy might alter the electrostatic potential surrounding the bound N(-)(3); the second, involving residues of helices B and G which are far from the haem iron atom, thus implying a long range effect on the bound N(-)(3). Based on the evidence that no significant conformational changes are found in the three-dimensional structures of N(-)(3)-free and N(-)(3)-bound myoglobin and on previous results on N(-)(3) binding to ferric myoglobin mutants in CD3 positions, we favour the first hypothesis, suggesting that the functional heterotropic modulation of monomeric myoglobin is mainly depending on a decrease of the positive charge density induced by the binding of anions to the haem distal side. PMID- 11904229 TI - Cold-induced enzyme inactivation: how does cooling lead to pyridoxal phosphate aldimine bond cleavage in tryptophanase? AB - The phenomenon of cold scission or cold lability, which entails a widespread variety of oligomeric enzymes, is still enigmatic. The effect of cooling on the activity and the quaternary structure of the pyridoxal 5'-phosphate (PLP) dependent enzyme, tryptophanase (Tnase), was studied utilizing single photon counting time-resolved spectrofluorometry. Upon cooling of holo-wild-type (wt) Tnase and its W330F mutant from 25 degrees C to 2 degrees C, a reduction in PLP fluorescence lifetime and rotational correlation time as well as inactivation and dissociation from tetramers to dimers were observed for both enzymes. Fluorescence anisotropy invariably decreased as a consequence of cooling, whether it was accompanied by a slight decrease in activity without significant dissociation, or by a substantial decrease in activity that was associated with either a partial or major dissociation. These results support the suggested conformational change that precedes the PLP-aldimine bond scission. It is proposed that cold inactivation is initiated by the weakening of hydrophobic interactions, leading to conformational changes which are the driving force for the aldimine bond cleavage. PMID- 11904231 TI - Structure-function relationship of three neurotoxins from the venom of Naja kaouthia: a comparison between the NMR-derived structure of NT2 with its homologues, NT1 and NT3. AB - Three homologous short-chain neurotoxins, named NT1, NT2 and NT3, were purified from the venom of Naja kaouthia. NT1 has an identical amino acid sequence to cobrotoxin from Naja naja atra [Biochemistry 32 (1993) 2131]. NT3 shares the same sequence with cobrotoxin b [J. Biochem. (Tokyo) 122 (1997) 1252], whereas NT2 is a novel 61-residue neurotoxin. Tests of their physiological functions indicate that NT1 shows a greater inhibition of muscle contraction induced by electrical stimulation of the nerve than do NT2 and NT3. Homonuclear proton two-dimensional NMR methods were utilized to study the solution tertiary structure of NT2. A homology model-building method was employed to predict the structure of NT3. Comparison of the structures of these three toxins shows that the surface conformation of NT1 facilitates the substituted base residues, Arg28, Arg30, and Arg36, to occupy the favorable spatial location in the central region of loop II, and the cation groups of all three arginines face out of the molecular surface of NT1. This may contribute greatly to the higher binding of NT1 with AchR compared to NT2 and NT3. PMID- 11904232 TI - The 11 beta-hydroxysteroid dehydrogenase type 2 activity in human placental microsomes is inactivated by zinc and the sulfhydryl modifying reagent N ethylmaleimide. AB - Proper glucocorticoid exposure in utero is vital to normal fetal organ growth and maturation. The human placental 11 beta-hydroxysteroid dehydrogenase type 2 enzyme (11 beta-HSD2) catalyzes the unidirectional conversion of cortisol to its inert metabolite cortisone, thereby controlling fetal exposure to maternal cortisol. The present study examined the effect of zinc and the relatively specific sulfhydryl modifying reagent N-ethylmaleimide (NEM) on the activity of 11 beta-HSD2 in human placental microsomes. Enzyme activity, reflected by the rate of conversion of cortisol to cortisone, was inactivated by NEM (IC(50)=10 microM), while the activity was markedly increased by the sulfhydryl protecting reagent dithiothreitol (DTT; EC(50)=1 mM). Furthermore, DTT blocked the NEM induced inhibition of 11 beta-HSD2 activity. Taken together, these results suggested that the sulfhydryl (SH) group(s) of the microsomal 11 beta-HSD2 may be critical for enzyme activity. Zn(2+) also inactivated enzyme activity (IC(50)=2.5 microM), but through a novel mechanism not involving the SH groups. In addition, prior incubation of human placental microsomes with NAD(+) (cofactor) but not cortisol (substrate) resulted in a concentration-dependent increase (EC(50)=8 microM) in 11 beta-HSD2 activity, indicating that binding of NAD(+) to the microsomal 11 beta-HSD2 facilitated the conversion of cortisol to cortisone. Thus, this finding substantiates the previously proposed concept that a compulsorily ordered ternary complex mechanism may operate for 11 beta-HSD2, with NAD(+) binding first, followed by a conformational change allowing cortisol binding with high affinity. Collectively, the present results suggest that cellular mechanisms of SH group modification and intracellular levels of Zn(2+) may play an important role in regulation of placental 11 beta-HSD2 activity. PMID- 11904233 TI - The intracerebral administration of phenytoin using controlled-release polymers reduces experimental seizures in rats. AB - PURPOSE: An alternative strategy for the treatment of intractable seizures may be to administer anticonvulsants directly into the brain near the site of a seizure focus using controlled-release polymers. We describe the pharmacokinetics of a phenytoin-ethylene-vinyl acetate (EVAc) controlled-release polymer and report the reduction of seizures in a cobalt-induced rat model of epilepsy with the intracerebral delivery of phenytoin using surgically implanted polymers. METHODS: In the pharmacokinetics study, the drug release rate of 50%-loaded phenytoin-EVAc polymers (n=3) was determined in vitro over 15 weeks initially and then several months later (over a 2-week period after 1 year of in vivo release). In the efficacy study, 85 rats underwent implantation of skull-mounted cortical electrodes for electrocorticography (ECoG) and then underwent application of cobalt chloride to the cerebral cortex for the induction of seizures. Rats in the treatment group (n=9) underwent surgical implantation of phenytoin-EVAc polymers and rats in the control group (n=10) underwent implantation of empty EVAc polymers. In the morbidity study, the potential histologic pathology of the intracerebral delivery of increasing doses of phenytoin from the polymer (10, 20, 30, and 50% loading) was assessed. RESULTS: Phenytoin was released in vitro from EVAc polymers in a controlled fashion with an initial release of 0.20% of the total loaded dose per week and a continued release of 0.70% of the total loaded dose per week after 365 days of implantation in the brain. The intracerebral controlled-release of phenytoin resulted in a statistically significant reduction in seizure activity in the treatment group as evidenced by lower Racine scores. The four groups of rats (n=5 per group) that underwent intracerebral implantation of 10, 20, 30, or 50%-loaded phenytoin-EVAc polymers displayed expected average weight gain and normal behavior over 365 days. One rat in the 50% group, however, died 354 days after polymer implantation for undetermined reasons. CONCLUSIONS: The intracerebral delivery of phenytoin using an EVAc polymer, which will release this drug for a calculated period of 3.5 years, resulted in a significant reduction in seizures in a rat model of cobalt-induced epilepsy by both behavioral and ECoG criteria. In rats, the long-term interstitial delivery of phenytoin in the brain was not associated with any deleterious effects. PMID- 11904234 TI - Mitochondrial dysfunction associated with neuronal death following status epilepticus in rat. AB - Status epilepticus (SE) in humans and animal models results in significant cerebral damage and an increased risk of subsequent seizures, associated with a characteristic pattern of neuronal loss particularly affecting the hippocampus. Seizure related cell death is considered to be excitotoxic, but studies have been limited, concentrating on terminal events rather than initial mechanisms. We have studied the biochemical events in the first few days following SE. Self sustaining limbic SE was induced in adult rats using perforant path stimulation, and animals were allowed to recover. Biochemical studies were performed at 16, 44 h and 8 days following SE, using spectrophotometric enzyme assays and HPLC on regional brain homogenates compared with those from sham-operated controls. Haematoxylin and eosin histology was also undertaken at each time point. Brain aconitase and alpha-ketoglutarate dehydrogenase (alphaKDH) activity were both significantly (P<0.05) reduced by approximately 20% in the first 16-44 h following status, but had returned to normal by 8 days. These enzymes are part of the tri-carboxylic acid (Krebbs) cycle in the mitochondrial matrix, and are known to be sensitive to free radical, especially peroxynitrite damage. There was a similar decrease in reduced glutathione levels. Histological studies confirmed evidence of acute neuronal damage up to 44 h, and neuronal loss by 8 days. This is the first in vivo demonstration of this pattern of mitochondrial dysfunction and loss of brain glutathione following SE. The pattern of abnormalities is consistent with reversible mechanisms being involved in excitotoxic cell damage. This, together with the timing of changes, suggests new avenues for therapeutic intervention. PMID- 11904235 TI - Linkage analysis between childhood absence epilepsy and genes encoding GABAA and GABAB receptors, voltage-dependent calcium channels, and the ECA1 region on chromosome 8q. AB - Childhood absence epilepsy (CAE) is an idiopathic generalised epilepsy (IGE) characterised by onset of typical absence seizures in otherwise normal children of school age. A genetic component to aetiology is well established but the mechanism of inheritance and the genes involved are unknown. Available evidence suggests that mutations in genes encoding GABA receptors or brain expressed voltage-dependent calcium channels (VDCCs) may underlie CAE. The aim of this work was to test this hypothesis by linkage analysis using microsatellite loci spanning theses genes in 33 nuclear families each with two or more individuals with CAE. Seventeen VDCC subunit genes, ten GABA(A)R subunit genes, two GABA(B) receptor genes and the ECA1 locus on 8q24 were investigated using 35 microsatellite loci. Assuming locus homogeneity, all loci gave statistically significant negative LOD scores, excluding these genes as major loci in the majority of these families. Positive HLOD scores assuming locus heterogeneity were observed for CACNG3 on chromosome 16p12-p13.1 and the GABRA5, GABRB3, GABRG3 cluster on chromosome 15q11-q13. Association studies are required to determine whether these loci are the site of susceptibility alleles in a subset of patients with CAE. PMID- 11904236 TI - Mutation (Ser284Leu) of neuronal nicotinic acetylcholine receptor alpha 4 subunit associated with frontal lobe epilepsy causes faster desensitization of the rat receptor expressed in oocytes. AB - To date five mutations in two major constituents of neuronal nicotinic acetylcholine receptor (nAChR) in the brain, i.e. alpha4 and beta2 subunits, have been identified to be associated with autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). Among them, only Ser284Leu, a point mutation in alpha4 subunit identified in ADNFLE as well as in a sporadic case with nocturnal frontal lobe epilepsy, remains to be characterized electrophysiologically. We examined the properties of rat nAChR harboring Ser284Leu reconstituted on Xenopus oocytes. Currents elicited in response to application of acetylcholine to oocytes expressing wild type or mutant nAChR were measured by a standard two microelectrode voltage clamp method. Compared with wild-type nAChR, the mutant nAChR had a comparable EC(50) value for acetylcholine whereas it showed faster desensitization and lower Cs(+)/Na(+) permeability ratio. Ser284Phe, a putative mutation constructed for comparison, exhibited similar properties. These findings indicate that Ser(284) plays an important role in gating of nAChR along with Thr(276) and Ser(280), and suggest that mutation at Ser(284) could reduce nAChR activity similar to other mutations of alpha4 subunit found in ADNFLE. PMID- 11904237 TI - Development of a new seizure severity questionnaire: initial reliability and validity testing. AB - PURPOSE: This report describes the initial steps for development of a new scale to assess seizure severity as a treatment response. METHODS: Standard methodology was used to develop the test instrument. Item generation was performed by selecting items from other questionnaires, and asking patients and epileptologists about seizure components. Face and content validity were assessed in a pilot study with patients and observers. The questionnaire was formatted as a structured interview for a reliability study. Construct validity was assessed with three existing questionnaires. Inter-rater reliability and test-retest reliability were performed as two independent ratings on one day, and one re interview. RESULTS: Based on item generation and pilot testing (33 patients, 28 observers), the Seizure Severity Questionnaire (SSQ) was organized into warning, activity-movement, and recovery (cognitive, emotional, and physical aspects) stages of seizures. Questions reviewed duration, severity, bothersomeness and overall ratings, and the most bothersome aspect of seizures. The mean SSQ Summary Score was 5.78+/-3.24, inter-rater reliability was 0.76 (N=91), and test-retest reliability was 0.74 (N=63). Construct validity showed statistically significant correlations with other scales. CONCLUSION: This study has explored the psychometric properties of the SSQ for face and content validity, inter-rater and test-retest reliability, and construct validity. The Summary Score reliably represents the major components of seizures. PMID- 11904238 TI - Ictal electrocorticographic findings related with surgical outcomes in nonlesional neocortical epilepsy. AB - PURPOSE: To characterize ictal electrocorticographic features related to surgical outcomes in nonlesional neocortical epilepsy (NE). METHODS: We analyzed 187 ictal electrocorticograms (ECoG) obtained from 18 patients who had undergone presurgical evaluation and subsequent neocortical resections (frontal: seven, parietal: one, occipital: four, multilobar: six). None of them had any MRI detectable lesions. Various ECoG data sets recorded from eight patients who achieved a favorable surgical outcome (either seizure free or more than 90% reduction of seizure frequencies) were compared with that from ten patients with unfavorable outcome (less than 90% reduction of seizure frequencies) (follow up duration: 47+/-11 months). RESULTS: Reproducible ictal onset zone (IOZ) in recurrent seizures (P=0.013) and persistent ictal discharges in IOZ from the onset to the end of seizure (P=0.004) were found more frequently in the patients with good outcome. Ictal onset patterns consisting of low voltage fast or high amplitude beta spikes predicted a good surgical outcome while rhythmic sinusoidal activity or rhythmic spike/sharp waves of slow frequency were predictive of poor outcome (P=0.01). The ictal onset rhythm consisting of gamma or beta frequencies was more prevalent in the favorable group (P=0.015). CONCLUSIONS: The presence of stable ictal circuit suggested by the consistent earliest activation of specific electrodes in the repetitive seizures (reproducible IOZ) and the active participation of IOZ throughout the attack were valuable prognostic factors in addition to the morphology and frequency of ictal onset rhythm. PMID- 11904239 TI - The costs of epilepsy in Italy: a prospective cost-of-illness study in referral patients with disease of different severity. AB - PURPOSE: [corrected] Epilepsy poses a considerable economic burden on society. However, information is insufficient on the comparative costs of different disease varieties. The purpose of this study was to compare the direct costs of epilepsy in referral patients with disease of different severity and duration. METHODS: Patients with newly diagnosed epilepsy (NDE), seizure remission (SR), occasional seizures (OS), frequent non-drug-resistant (NDR) and drug-resistant (DR) seizures, and surgical candidates (SC) from 14 epilepsy centers were the target population. All patients were followed prospectively for 12 months and all medical and paramedical contacts for diagnostic and therapeutic services were noted with details, using ad-hoc diaries and semistructured questionnaires. RESULTS: The study population comprised 525 consecutive children and adults with partial (68%), generalized (25%) and undetermined epilepsy (4%) as follows: NDE 70; SR 131; OS 108; NDR 101; DR 107; SC 8. Ambulatory visits (mean 2.8 per patient per year) were the leading service in all groups, followed by EEG recordings (1.8) and biochemical assays (1.1). At entry, the commonest drugs were carbamazepine (50%), valproate (37%), phenobarbital (21%), vigabatrin (14%) and lamotrigine (11%). New antiepileptic drugs (AED) were used increasingly with the severity of the disease. The total annual costs varied significantly across groups: 3945 Euro (SC), 2198 Euro (DR), 1626 Euro (NDR), 1002 Euro (NDE), 558 Euro (OS), 412 Euro (SR). The main item of expenditure was hospital stay (including day-hospital), followed by drug treatment and outpatient visits. The costs of outpatient services, hospital services and drugs varied significantly across groups. CONCLUSIONS: The direct costs of epilepsy vary significantly depending on the severity of the disease and the response to treatment. Hospital admissions and drugs are the commonest items of expenditure. PMID- 11904240 TI - Carbamazepine toxicity during combination therapy with levetiracetam: a pharmacodynamic interaction. AB - Levetiracetam is a novel antiepileptic drug with an unknown mechanism of action. To-date levetiracetam is not known to be associated with any clinically significant pharmacokinetic interaction. Similarly, levetiracetam has not been associated with any pharmacodynamic interactions. We present four patients with severe refractory epilepsy in whom introduction of levetiracetam led to disabling symptoms compatible with carbamazepine toxicity requiring either carbamazepine dose reduction or levetiracetam withdrawal. As carbamazepine and carbamazepine epoxide blood levels were not altered during levetiracetam co-medication, a pharmacodynamic interaction is suggested. Therefore, during levetiracetam co medication with carbamazepine, patients should be monitored closely for symptoms of carbamazepine toxicity. PMID- 11904241 TI - The ketogenic diet in children, adolescents and young adults with refractory epilepsy: an Italian multicentric experience. AB - PURPOSE: This collaborative study by three Italian groups of child neuropsychiatrists was carried on to evaluate the efficacy and safety of the classic 4:1 ketogenic diet as add-on treatment in refractory partial or generalized epilepsy in children, adolescents and young adults. METHODS: We performed a prospective add-on study in 56 refractory epilepsy young patients (age 1-23 years, mean 10.4 years), all with both symptomatic and cryptogenic, generalized or partial epilepsies. Child neuropsychiatrists worked with nutritional team for sample selection and patients management. The ketogenic diet was added to the baseline antiepileptic drugs and the efficacy was rated according to seizure type and frequency. During treatment, seizure frequency, side effects, urine and blood ketone levels and other parameters were systematically evaluated. RESULTS: Patients have been treated for 1-18 months (mean 5 months). A >50% reduction in seizure frequency was gained in 37.5 and 26.8% of patients after 3 and 6 months, respectively, at 12 months, this number fell by 8.9%. No significant relationship between diet efficacy and seizure or epilepsy type, age at diet onset, sex and etiology of epilepsy was noted. Nevertheless, it seems noteworthy that 64% of our patients with neuronal migration disorders improved on this diet. Adverse effects occurred, mainly in the first weeks of treatment, in 32 patients (57.1%), but were generally mild and transient. In seven patients (12.5%) it was possible to withdraw one to two AED after 3-4 months on ketogenic diet. CONCLUSION: This initial experience with the ketogenic diet was effective in difficult-to-treat patients with partial and generalized epilepsies, though its efficacy dropped significantly by 9-12 months. PMID- 11904243 TI - Personal health records on the internet: a snapshot of the pioneers at the end of the 20th Century. AB - Internet-based, personal health records have the potential to profoundly influence the delivery of health care in the 21st Century, by changing the loci and ownership of the record from one that is distributed among the various health care providers a patient has seen in his lifetime, to one with a single source that is accessible from anywhere in the world and under the shared ownership and control of the patient and his provider(s). Internet-based personal health records (PHRs) include any internet-accessible application that enables a patient (or his guardian, the 'mom') to review, annotate, create or maintain a record of any aspect(s) of his health condition, medication, medical problems, allergies, vaccination history, visit history or communications with his healthcare providers. The current state-of-the-art for personal health records is best characterized as 'beta releases'. As the field matures and gains more experience, these applications will improve significantly in ease of use and functionality. PMID- 11904242 TI - Memory reorganization in adult brain: observations in three patients with temporal lobe epilepsy. AB - We present three patients with left-sided temporal lobe epilepsy who exhibited preoperatively a neuropsychological pattern characteristic for interhemispheric language transfer (marked nonverbal memory deficits, relatively preserved verbal memory and language performance). The Wada test indicated atypical language dominance in two patients, but one patient was clearly left hemispheric language dominant. All patients showed a marked recovery of nonverbal memory after left sided surgery. Results are discussed with respect to memory transfer and plasticity for memory functions in the adult brain. PMID- 11904244 TI - A Delphi technique as a method for selecting the content of an electronic patient record for asthma. AB - INTRODUCTION: An electronic patient record (EPR) with disease-specific data may support improvement of the quality of care for patients with chronic diseases. The structure and content of such a record can only be assessed by clinicians in co-operation with IT-specialists, because, the result has to be clinically relevant, easily accessible and adjusted to the information needs of different workers in primary care. METHODS: We applied a modified Delphi-procedure--a method characterized by anonymous written comments by an expert panel. The panel had to agree about the question whether or not an item should be included in the EPR. The questions for the written comments were prepared by a steering committee (general practitioners (GP) and health scientists, either expert in asthma and disease management or IT-specialist) based on the guidelines for diagnosing and treating asthma of the Dutch College of General Practitioners (DCGP). When agreement within the panel was < or = 70%, we sent a modified format to the expert panel for reassessment. RESULTS: Three written commentary rounds were necessary, in which 95 potential items were discussed with the expert panel. In the first round they selected 50 items relating to diagnosing asthma and 22 concerning the treatment of asthma. During the second round 17 items were still under discussion and six were rejected. In subsequent rounds, the expert-panel assessed the best registration format (operationalisation). The written rounds failed to create a full consensus. Therefore the study ended with a consensus meeting of the expert panel. Due to the presence and contribution of nearly all experts, consensus could be reached about the structure and contents of the EPR on asthma. DISCUSSION: The modified Delphi procedure, proved to be a feasible method for selecting the optimal content of an electronic registration protocol. Both, written and verbal commentary rounds were necessary. The existence of a set of guidelines was essential. PMID- 11904245 TI - Manual semantic tagging to improve access to information in narrative electronic medical records. AB - OBJECTIVE: To assess effectiveness of information retrieval from narrative patient records utilizing manually supplied semantic tags. DESIGN: Retrospective evaluation of narrative electronic record notes with respect to consistency of manually supplied semantic tags. Assessment of retrieval effectiveness using string matching methods and methods based on structural characteristics of data. SETTING: Department of Neurology in a Norwegian university hospital using a document based electronic patient record system offering template assisted manual semantic indexing of textual record notes. MEASUREMENTS: Proportion of consistently tagged information. Retrieval effectiveness expressed as recall (sensitivity) and precision (positive predictive value). RESULTS: Seventy-five percent of the content was consistently tagged. Retrieval models based on simple string matching performed better than models based on semantic tags alone with respect to best mean recall and mean precision at high recall-levels. Models combining both simple string matching and semantic tagging performed better than separate models with respect to best mean recall (0.95) and mean precision at high levels of recall. CONCLUSION: Although template assisted semantic indexing of narrative electronic patient record notes showed suboptimal consistency, it significantly improved retrieval effectiveness with respect to best mean recall obtained by individual strategies and with respect to precision at any recall level. PMID- 11904246 TI - Master of science program in health information management at Heidelberg/Heilbronn: a health care oriented approach to medical informatics. AB - In the year 2000 the University of Heidelberg and the University of Applied Sciences Heilbronn established a second educational program in medical informatics, leading to a Master of Science (MSc) degree. In addition to their 4.5 year medical informatics program as an informatics-based approach to medical informatics, a postgraduate program in 'health information management' (Informationsmanagement in der Medizin) was set up as a complementary health care oriented approach to this field. The aim of the MSc program is to qualify physicians and other health care professionals to work as medical informaticians, particularly in the area of health information management. We admit 15 new students into the program each year. The intended program length is 15 months, comprising two study semesters (14 weeks per semester) and three months for the Master's thesis. The graduates are awarded the title 'Master of Science' by the Medical Faculty of the University of Heidelberg. The program is part of the International Partnership in Health Informatics Education of the Universities of Amsterdam, Heidelberg/Heilbronn, Minnesota and Utah. We report on this new program and on our experience with our very first students. The curriculum is compared with the related MSc programs. PMID- 11904247 TI - Physicians' attitudes towards the computerization of clinical practice in Hong Kong: a population study. AB - OBJECTIVE: To identify the prevailing attitudes among physicians to use of computers in the clinical setting and specifically those attitudes that may be associated with the adoption of computers in practice. DESIGN: A self-completed, 20-question postal questionnaire sent to 4850 randomly selected physicians. The questionnaire focused on details of the physicians' practice; actual computerization of or intention to computerize clinical and administrative functions; attitudes towards computerization; self-perceived computer ability and knowledge; and demographic information. The attitude statements were grouped under four themes according to a factor analysis. RESULTS: The survey was completed by 897 physicians. Only physicians in 'individual' practices were included in the analysis. This group was further dichotomized into clinical users (those with one or more clinical functions computerized) and non-clinical users. Non-clinical users were older and had fewer specialist qualifications. Although there was strong support for the attitude statements among both groups with regard to the benefit of computerization to patient care, there was much less support for electronic medical records. Non-clinical users were concerned about the potentially negative impact of computerization on the clinical encounter and the perceived high cost of computerization. DISCUSSION: The attitudes among current clinical users and non-users were substantially different. The most important disincentives to computerization were the potential for interference with the patient-physician encounter and the cost of computerizing multiple practice locations. Turning these disincentives into opportunities for change remains the challenge. PMID- 11904248 TI - Metadata tables to enable dynamic data modeling and web interface design: the SEER example. AB - A wealth of information addressing health status, outcomes and resource utilization is compiled and made available by various government agencies. While exploration of the data is possible using existing tools, in general, would-be users of the resources must acquire CD-ROMs or download data from the web, and upload the data into their own database. Where web interfaces exist, they are highly structured, limiting the kinds of queries that can be executed. This work develops a web-based database interface engine whose content and structure is generated through interaction with a metadata table. The result is a dynamically generated web interface that can easily accommodate changes in the underlying data model by altering the metadata table, rather than requiring changes to the interface code. This paper discusses the background and implementation of the metadata table and web-based front end and provides examples of its use with the NCI's Surveillance, Epidemiology and End-Results (SEER) database. PMID- 11904249 TI - Different lenses, improved outcomes: a new approach to the analysis and design of healthcare information systems. AB - Healthcare systems are complex sociotechnical systems in which many information system innovations fail because of problems in planning or design. One of the reasons for this is that traditional analysis methods were designed for stable, relatively simple systems and single users. New analytical approaches are needed that can encompass the complexity of changing systems and multiple, interacting users. The author suggests that integrating two seemingly disparate approaches may be helpful in achieving successful designs for the complexities of healthcare systems. The first is derived from Carper's four ways of knowing; the second is Cognitive Work Analysis (CWA). The utility of the approach is demonstrated via a case study. PMID- 11904250 TI - Emergency department drug therapy for status epilepticus in adults. PMID- 11904252 TI - Emergency analgesia in the paediatric population. Part II Pharmacological methods of pain relief. PMID- 11904253 TI - Training in intensive care medicine: an accident and emergency trainee's perspective. PMID- 11904254 TI - Rapid sequence induction in the emergency department: a strategy for failure. AB - BACKGROUND: Rapid sequence induction (RSI) is increasingly used by emergency physicians in the emergency department. A feared complication of the technique is the inability to intubate and subsequently ventilate the patient. Current drills based on anaesthetic practice may be unsuitable for use in the emergency department. OBJECTIVE: To construct a drill for failed adult intubation in the emergency department. METHODS: Literature review and consensus knowledge. RESULTS: A drill for failed adult intubation in the emergency department is given. SUMMARY: Failure to intubate following RSI in the emergency department is a feared complication. Practitioners must have a predetermined course of action to cope with this event. The guidelines presented here are tailored for use by the emergency physician. PMID- 11904255 TI - Choice of fluid for resuscitation of septic shock. AB - OBJECTIVES: To determine current practice in choice of fluid resuscitation in children following publication of a systematic review that demonstrated a higher mortality in patients treated with human albumin solution. METHODS: A descriptive telephone and postal questionnaire survey directed at the on call paediatric registrar, lead clinician for paediatrics and the paediatric pharmacist at each of 33 hospitals within the Greater London area. The study was coordinated by the Paediatric Intensive Care Unit at St Mary's Hospital, London. The questionnaire was designed to assess whether a protocol/guidelines existed for resuscitation fluid in children with septic shock; whether the participants were aware of the systematic review and if so, had it changed clinical practice. The word "protocol" was used in its broadest sense to include guideline and policy. RESULTS: 11 hospitals had guidelines for fluid resuscitation of septic shock in children. These varied greatly: only three gave clear instructions of which fluid to use and how to use it. Choice of fluid varied widely and there was wide discrepancy between consultant's and registrar's choice of fluid. The systematic review had lead to a change in policy in two thirds of respondents. CONCLUSION: It is apparent that few paediatric departments have a written protocol or guidelines for the management of septic shock that is accessible to all those concerned in the acute treatment of seriously ill children. The systematic review into choice of fluid has had an impact on clinical practice with no data regarding whether this is in the patient's best interests. PMID- 11904256 TI - A prospective, observational study of a chest pain observation unit in a British hospital. AB - OBJECTIVES: To establish a chest pain observation unit, monitor its performance in terms of appropriate discharge after assessment, and estimate the cost per patient. METHODS: Prospective, observational, cohort study of patients attending a large, city, teaching hospital accident and emergency department between 1 March 1999 and 29 February 2000 with acute undifferentiated chest pain. Patients were managed on a chest pain observation unit, entailing two to six hours of observation, serial electrocardiograph recording, cardiac enzyme measurement, and, where appropriate, exercise stress test. Patients were discharged home if all tests were negative and admitted to hospital if tests were positive or equivocal. The following outcomes were measured-proportion of participants discharged after assessment; clinical status three days after discharge; cardiac events and procedures during the following six months; and cost of assessment and admission. RESULTS: Twenty three participants (4.3%) had a final diagnosis of myocardial infarction. All were detected and admitted to hospital. A total of 461 patients (86.3%) were discharged after assessment, 357 (66.9%) avoided hospital admission entirely. At review three days later these patients had no new ECG changes and only one raised troponin T measurement. In the six months after assessment, three cardiac deaths, two myocardial infarctions, and two revascularisation procedures were recorded among those discharged. The mean cost of assessment and hospital admission was 221 pound per patient, or 323 pound if subsequent interventional cardiology costs were included. CONCLUSIONS: The chest pain observation unit is a practical alternative to routine care for acute chest pain in the United Kingdom. Negative assessment effectively rules out immediate, serious morbidity, but not longer term morbidity and mortality. Costs seem to be similar to routine care. PMID- 11904257 TI - Emergency management of chest pain: patient satisfaction with an emergency department based six hour rule out myocardial infarction protocol. AB - OBJECTIVE: To measure the level of patient satisfaction with an emergency department based chest pain assessment unit. DESIGN: Structured patient satisfaction surveys. SETTING: Inner city university hospital emergency department. PARTICIPANTS: 383 consecutive patients aged over 25 years with probable cardiac chest pain of less than 12 hours duration at moderate to low risk of acute myocardial infarction. INTERVENTION: Two structured questionnaires the first addressing satisfaction with different aspects of the health care process, the second designed to assess global satisfaction outcomes. RESULTS: 274 patients (74%) fully or partly answered the first questionnaire. There were high levels of satisfaction with all process of care issues (waiting time, information, discussion, explanation, pain management, personal needs, family needs, and discharge preparation). Altogether 258 patients fully or partly answered the second questionnaire. Global satisfaction was high. Subgroup data analysis showed white patients significantly more satisfied than non-white patients (p<0.0001), and over 45s significantly more satisfied than under 45s (p<0.01). A number of issues were raised in the free comment section of the second questionnaire. The lack of a definitive diagnosis at discharge was a recurring theme. CONCLUSION: Chest pain assessment units are acceptable from a patient perspective. PMID- 11904258 TI - What's the point of ST elevation? AB - OBJECTIVE: The magnitude of ST elevation is a key piece of information in the decision to thrombolyse in acute myocardial infarction. The ability of clinicians to reliably identify ST elevation has not been previously assessed. This study sought to determine the variability in assessment of ST elevation in a group of doctors who commonly prescribe thrombolysis. METHODS: The study was conducted in three large teaching hospitals in Manchester, England. A convenience sample of 63 SHOs and SpRs from emergency and general medicine were recruited. Each was shown three sample ECG complexes. They were asked to identify and quantify the degree of ST elevation. They then indicated the points on the ECG from which they measured ST elevation. RESULTS: ST elevation was not identified in 12% of cases. Doctors used a wide variety of points on the ST segment to assess elevation, this resulted in a wide variation in the observed magnitude of ST elevation. CONCLUSION: No guidance exists on where exactly ST elevation should be measured. This study shows a wide variation in practice. Protocol led thrombolysis decision pathways may be compromised by these findings. PMID- 11904259 TI - The electrocardiographic differential diagnosis of ST segment depression. PMID- 11904260 TI - Accident and emergency department access to the child protection register: a questionnaire survey. AB - OBJECTIVES: To ascertain how UK accident and emergency (A&E) departments access the child protection register, their levels of satisfaction with that access and their criteria for checking the register. METHODS: A postal questionnaire was sent to 254 "major" A&E departments listed in the 1996 British Association for Accident and Emergency Medicine directory. RESULTS: 190 questionnaires were returned (response rate 75%). Ninety (48%) responding departments access the register through the duty social worker, 33 (17%) use a computerised copy, 32 (17%) a hard copy and 27 (14%) a combination. Twenty seven of 33 respondents (82%) using a computerised copy were satisfied with their mode of access. This compares with figures of 21 (66%) for hard copy, 45 (50%) for duty social worker and 14 (50%) for a combination. No departments using the duty social worker checked all patients routinely compared with 23 (72%) for hard copy, 22 (67%) for computer copy and 12 (44%) for departments using a combination of modes of access. CONCLUSION: There is no uniformity of the way in which UK A&E departments access the child protection register and there is also substantial variation in the criteria used to check the register. This survey suggests that the most common form of access (via the duty social worker) often fails to meet the needs of A&E departments, principally because it takes so long. PMID- 11904261 TI - Acceptability of asking patients about violence in accident and emergency. AB - OBJECTIVES: To assess the acceptability to patients attending accident and emergency (A&E) of routine questioning about violence. METHODS: A questionnaire survey (15 questions; 5 point Likert scale) was distributed to a representative sample of all adult patients attending a district general hospital A&E department, Lancashire, England over a seven day period. RESULTS: 303 questionnaires were distributed and 281 returned questionnaires were available for analysis. Some 67% (95%CI 60% to 74%) of patients agreed that people attending A&E should routinely be asked about whether they have been assaulted. Altogether 89% (95%CI 85% to 93%) thought that health care staff should encourage victims of abuse or violence to inform the police, while 74% (95%CI 68% to 80%) thought that health care staff should routinely inform the police. While only 45% (95%CI 36% to 54%) of patients thought that people who had been assaulted would be likely to tell if asked, 81% (95%CI 76% to 86%) thought that if they themselves were victims they would tell if asked directly. CONCLUSIONS: Patients attending A&E departments support routine questioning by doctors and nurses about violence. They also support health professionals routinely informing the police in cases of violence. Further research is required into the outcomes of routine and direct questioning in A&E of patients about their exposure to violence. PMID- 11904262 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Reimplantation of the nail root in fingertip crush injuries in children. AB - A short cut review was carried out to establish whether reimplantation of the nail improved cosmetic outcome after crush injury to the fingertip in children. Altogether 35 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 11904264 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Antibiotics in non-venomous snakebite. AB - A short cut review was carried out to establish whether prophylactic antibiotics reduced the incidence of infection after non-venemous snake bite. Altogether 60 papers were found using the reported search, of which two presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 11904265 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Intra-articular lidocaine for acute anterior shoulder dislocation reduction. AB - A short cut review was carried out to establish how intra-articular lidocaine compared with intravenous analgesia and sedation during reduction of anterior shoulder dislocations. Altogether 146 papers were found using the reported search, of which three presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 11904266 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Propofol for resistant status epilepticus. AB - A short cut review was carried out to establish whether propofol is effective at stopping fitting in resistant status epilepticus. Altogether 24 papers were found using the reported search, of which six presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 11904267 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Regional nerve block in fractured neck of femur. AB - A short cut review was carried out to establish whether regional nerve block is better than intravenous analgesia in reducing pain in hip fractures. Altogether 21 papers were found using the reported search, of which four presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 11904268 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Bronchodilator delivery in acute severe asthma in adults. PMID- 11904269 TI - Article 7. Money, money, money. (Where does it come from, how do we control it, and how far should we go to get more?). PMID- 11904270 TI - Do ambulance crews with one advanced paramedic skills officer have longer scene times than crews with two? AB - OBJECTIVE: In 1999, the Metropolitan Ambulance Service (MAS), Melbourne, Australia began implementing The Emergency Operations Plan (1998). One of the initiatives of the plan was the addition of crews with one advanced paramedic skills (APS) officer and one non-APS officer (mixed crews). All previous APS crews contained two APS officers working together. There was concern that mixed crews would have longer scene times than all-APS crews. This study aims to compare scene times at time critical cases for mixed crews and all-APS crews. METHOD: Prospective, non-randomised comparison of scene times for time critical cases for three mixed crew units and three all-APS units for the months of August to October 1999. The crew types were also compared by explicit retrospective audit for rates of APS procedures attempted and APS procedure failure rates. Data were analysed using SPSS, t test, and chi(2) test where appropriate. RESULTS: There were 1700 time critical cases in the study period of which 1537 had valid data for the calculation of scene times. A total of 714 cases were attended by mixed crews and 823 cases by all-APS crews. The mean scene time for mixed crews was 15.54 minutes compared with 16.92 minutes for all-APS crews. This difference is statistically significant (p=0.002). All-APS crews performed a slightly higher number of APS procedures (0.90/time critical case versus 0.76/time critical case; p=0.001). There was no significant difference in procedure failure rates. CONCLUSION: Mixed crews demonstrated shorter scene times than all-APS crews, although this is unlikely to be clinically significant. The concern that mixed crews would have longer scene time was not substantiated and should not be considered as a barrier to the development of mixed crew staffing models. PMID- 11904271 TI - Delphi type methodology to develop consensus on the future design of EMS systems in the United Kingdom. AB - OBJECTIVE: To develop consensus opinion on future design characteristics of Emergency Medical Services (EMS) systems in the UK with particular regard to advanced life support skills (ALS). DESIGN: A Delphi questionnaire design with two rounds to gain a consensus of opinion. Investigation of four aspects of EMS design is reported-type of response to a priority based dispatch category, transportation options, enhancement of paramedic skills, and structure of a first responder system. SUBJECTS: Chief executives, directors of operations, and medical directors of Ambulance Trusts in the United Kingdom. OUTCOME MEASURES: Likert scales (0-9) to score opinion on a series of statements with achievement of inter-round consistency. A median score of 0-4 was classified as disagreement and 6-9 as agreement. RESULTS: A 65% response to the first questionnaire and with iteration, 52% response to the second questionnaire was attained. A tiered response (paramedics, technicians, and basic life support first responders) with technicians responding to selected category A and B calls and all category C calls (median score (MS) 7.5, interquartile range (IQR) 4), was recommended. Inter-unit handover of selected calls to maintain paramedic availablity ( MS 7.5, IQR 3.75) and enhancement of paramedic skills (MS 7.0, IQR 4.0) was also proposed. Finally, the development of a first responder system fully integrated into the EMS (MS 8.0, IQR 2.75) involving other agencies including the police force, fire service, and trained members of the local community was agreed. CONCLUSIONS: Senior expert staff from Ambulance Trusts in the UK achieved consensus on certain design characteristics of EMS systems. These are significantly different from the present EMS model. PMID- 11904272 TI - The acceptability of an emergency medical dispatch system to people who call 999 to request an ambulance. AB - OBJECTIVE: To determine the acceptability of an emergency medical dispatch (EMD) system to people who call 999 to request an ambulance. METHODS: Postal questionnaires to two systematic random samples of approximately 500 named callers to one ambulance service before, and one year after, the introduction of EMD. RESULTS: The response rate was 72% (355 of 493) before, and 63% (297 of 466) after, EMD. There was a reduction, from 81% (284 of 349) to 70% (200 of 286), in the proportion of callers who found all the questions asked by the call taker relevant, although this did not adversely affect the proportion of callers who were very satisfied with the 999 call, which increased from 78% (268 of 345) to 86% (247 of 287). The proportion of callers who reported receiving first aid advice increased from 7% (23 of 323) to 43% (117 of 272) and general information from 13% (41 of 315) to 58% (157 of 269). Satisfaction levels with the amount of advice given increased, while satisfaction with response times remained stable at 76% (254 of 320) very satisfied before and 78% (217 of 279) after EMD. The proportion of respondents very satisfied with the service in general increased from 71% (238 of 336) to 79% (220 of 277). There was evidence in respondents' written comments of two potential problems with EMD from the caller's viewpoint. Firstly, some callers were advised to take actions that were subsequently not needed; secondly, a small number of callers felt that the ambulance crew did not treat the situation as seriously as they would have liked. CONCLUSIONS: Introducing EMD increases the amount of first aid and general advice given to callers, and satisfaction with these aspects of the service, while maintaining satisfaction with response times. Overall satisfaction with the service increased. However, some changes may be needed to prevent a small amount of dissatisfaction directly associated with EMD. PMID- 11904273 TI - Recovery from pH 6.38: lactic acidosis complicated by hypothermia. AB - Survival after extreme arterial acidosis is uncommon. A case of metformin induced lactic acidosis is described where the presenting pH was 6.38 exacerbated by hypothermia (29 degrees C). Increased anion gap acidosis, its varied aetiology, potential reversibility, and the role of hypothermia are discussed. Early liaison with a medical toxicology unit is recommended when this rare condition is suspected. PMID- 11904274 TI - Transient cervical neurapraxia associated with cervical spine stenosis. AB - A 43 year old woman presented with a history of a hyperextension cervical injury resulting in transient quadriplegia. Cervical spine radiography revealed developmental spinal stenosis and magnetic resonance imaging demonstrated underlying spinal cord oedema secondary to contusion, with a herniated disc at C3 C4. The Torg ratio may be used to aid the initial diagnosis of cervical spine stenosis. Indications for operative treatment of these patients are controversial and these patients should receive further expert assessment. PMID- 11904275 TI - Transient paraplegia as a presenting feature of aortic dissection in a young man. AB - This report describes a case of aortic dissection in a 32 year old man who presented with mild central chest pain and transient paraplegia. Complete recovery of paraplegia in aortic dissection has previously been reported but this is the first case report in which full resolution of the paraplegia occurred within half an hour of presentation. The case emphasises the importance of careful history taking and eliciting subtle clinical signs, which helped in arriving at the correct diagnosis in this unusual presentation. PMID- 11904276 TI - Tension pneumothoraces not responding to needle thoracocentesis. PMID- 11904277 TI - The use of transthoracic echocardiography to guide thrombolytic therapy during cardiac arrest due to massive pulmonary embolism. PMID- 11904278 TI - Surgical emphysema over the pelvis: an unusual physical sign found on primary survey. AB - Open fractures of the pelvis are associated with high energy trauma and present a challenge to successful management and sometimes, early and correct diagnosis. These patients require more aggressive blood resuscitation particularly in the first 24 hours, repeated wound care operations, and often require a diverting colostomy. Usually these pelvic fractures can be distinguished from closed pelvic fractures by an open wound or lacerations of the vagina and rectum. Occasionally, however, the wounds associated with these fractures may remain undetected and the severity of the injury underestimated until complications develop. The authors believe this to be the first report of subcutaneous surgical emphysema associated with an open pelvic fracture. PMID- 11904279 TI - Mediastinal emphysema after a minor oral laceration. PMID- 11904280 TI - What about patient satisfaction following acute ankle sprains? PMID- 11904281 TI - Calcium for hyperkalaemia in digoxin toxicity. PMID- 11904282 TI - Journal clubs in clinical medicine. PMID- 11904283 TI - Two cases of near asphyxiation in children, using non-releasing plastic garden ties. PMID- 11904284 TI - Tuberculous osteomyelitis. PMID- 11904285 TI - A misdiagnosed fracture of the calcaneum. PMID- 11904286 TI - Screening for alcohol misuse. PMID- 11904288 TI - A direct test of the reductionist approach to structural studies of calmodulin activity: relevance of peptide models of target proteins. AB - Ca(2+)-saturated calmodulin (CaM) directly associates with and activates CaM dependent protein kinase I (CaMKI) through interactions with a short sequence in its regulatory domain. Using heteronuclear NMR (13)C-(15)N-(1)H correlation experiments, the backbone assignments were determined for CaM bound to a peptide (CaMKIp) corresponding to the CaM-binding sequence of CaMKI. A comparison of chemical shifts for free CaM with those of the CaM. CaMKIp complex indicate large differences throughout the CaM sequence. Using NMR techniques optimized for large proteins, backbone resonance assignments were also determined for CaM bound to the intact CaMKI enzyme. NMR spectra of CaM bound to either the CaMKI enzyme or peptide are virtually identical, indicating that calmodulin is structurally indistinguishable when complexed to the intact kinase or the peptide CaM-binding domain. Chemical shifts of CaM bound to a peptide (smMLCKp) corresponding to the calmodulin-binding domain of smooth muscle myosin light chain kinase are also compared with the CaM. CaMKI complexes. Chemical shifts can differentiate one complex from another, as well as bound versus free states of CaM. In this context, the observed similarity between CaM. CaMKI enzyme and peptide complexes is striking, indicating that the peptide is an excellent mimetic for interaction of calmodulin with the CaMKI enzyme. PMID- 11904289 TI - Protection against anoikis and down-regulation of cadherin expression by a regulatable beta-catenin protein. AB - beta-Catenin signaling plays a key role in a variety of cellular contexts during embryonic development and tissue differentiation. Aberrant beta-catenin signaling has also been implicated in promoting human colorectal carcinomas as well as a variety of other cancers. To study the molecular and cellular biological functions of beta-catenin in a controlled fashion, we created a regulatable form of activated beta-catenin by fusion to a modified estrogen receptor (ER) ligand binding domain (G525R). Transfection of tissue culture cells with expression vectors encoding this hybrid protein allows the signal transduction function of beta-catenin to be induced by the synthetic estrogen, 4-hydroxytamoxifen, leading to regulated activation of a beta-catenin-lymphocyte enhancer-binding factor dependent reporter gene as well as induction of endogenous cyclin D1 expression. The activation of ER-beta-catenin signaling rescues RK3E cells from anoikis and correlates with an increased phosphorylation of mitogen-activated protein kinase. The inhibition of anoikis by ER-beta-catenin can be abolished by a mitogen activated protein kinase pathway inhibitor, PD98059. Evidence is also provided to show that ER-beta-catenin down-regulates cadherin protein levels. These findings support a key role for activated beta-catenin signaling in processes that contribute to tumor formation and progression. PMID- 11904290 TI - Proteomics analysis of cellular response to oxidative stress. Evidence for in vivo overoxidation of peroxiredoxins at their active site. AB - The proteomics analysis reported here shows that a major cellular response to oxidative stress is the modification of several peroxiredoxins. An acidic form of the peroxiredoxins appeared to be systematically increased under oxidative stress conditions. Peroxiredoxins are enzymes catalyzing the destruction of peroxides. In doing so, a reactive cysteine in the peroxiredoxin active site is weakly oxidized (disulfide or sulfenic acid) by the destroyed peroxides. Cellular thiols (e.g. thioredoxin) are used to regenerate the peroxiredoxins to their active state. Tandem mass spectrometry was carried out to characterize the modified form of the protein produced in vivo by oxidative stress. The cysteine present in the active site was shown to be oxidized into cysteic acid, leading to an inactivated form of peroxiredoxin. This strongly suggested that peroxiredoxins behave as a dam upon oxidative stress, being both important peroxide-destroying enzymes and peroxide targets. Results obtained in a primary culture of Leydig cells challenged with tumor necrosis factor alpha suggested that this oxidized/native balance of peroxiredoxin 2 may play an active role in resistance or susceptibility to tumor necrosis factor alpha-induced apoptosis. PMID- 11904291 TI - Phloxine B interacts with the cystic fibrosis transmembrane conductance regulator at multiple sites to modulate channel activity. AB - The fluorescein derivative phloxine B is a potent modulator of the cystic fibrosis transmembrane conductance regulator (CFTR). Low micromolar concentrations of phloxine B stimulate CFTR Cl(-) currents, whereas higher concentrations of the drug inhibit CFTR. In this study, we investigated the mechanism of action of phloxine B. Phloxine B (1 microm) stimulated wild-type CFTR and the most common cystic fibrosis mutation, DeltaF508, by increasing the open probability of phosphorylated CFTR Cl(-) channels. At each concentration of ATP tested, the drug slowed the rate of channel closure without altering the opening rate. Based on the effects of fluorescein derivatives on transport ATPases, these data suggest that phloxine B might stimulate CFTR by binding to the ATP-binding site of the second nucleotide-binding domain (NBD2) to slow the dissociation of ATP from NBD1. Channel block by phloxine B (40 microm) was voltage-dependent, enhanced when external Cl(-) concentration was reduced and unaffected by ATP (5 mm), suggesting that phloxine B inhibits CFTR by occluding the pore. We conclude that phloxine B interacts directly with CFTR at multiple sites to modulate channel activity. It or related agents might be of value in the development of new treatments for diseases caused by the malfunction of CFTR. PMID- 11904292 TI - Mlo, a modulator of plant defense and cell death, is a novel calmodulin-binding protein. Isolation and characterization of a rice Mlo homologue. AB - Transient influx of Ca(2+) constitutes an early event in the signaling cascades that trigger plant defense responses. However, the downstream components of defense-associated Ca(2+) signaling are largely unknown. Because Ca(2+) signals are mediated by Ca(2+)-binding proteins, including calmodulin (CaM), identification and characterization of CaM-binding proteins elicited by pathogens should provide insights into the mechanism by which Ca(2+) regulates defense responses. In this study, we isolated a gene encoding rice Mlo (Oryza sativa Mlo; OsMlo) using a protein-protein interaction-based screening of a cDNA expression library constructed from pathogen-elicited rice suspension cells. OsMlo has a molecular mass of 62 kDa and shares 65% sequence identity and scaffold topology with barley Mlo, a heptahelical transmembrane protein known to function as a negative regulator of broad spectrum disease resistance and leaf cell death. By using gel overlay assays, we showed that OsMlo produced in Escherichia coli binds to soybean CaM isoform-1 (SCaM-1) in a Ca(2+)-dependent manner. We located a 20 amino acid CaM-binding domain (CaMBD) in the OsMlo C-terminal cytoplasmic tail that is necessary and sufficient for Ca(2+)-dependent CaM complex formation. Specific binding of the conserved CaMBD to CaM was corroborated by site-directed mutagenesis, a gel mobility shift assay, and a competition assay with a Ca(2+)/CaM-dependent enzyme. Expression of OsMlo was strongly induced by a fungal pathogen and by plant defense signaling molecules. We propose that binding of Ca(2+)-loaded CaM to the C-terminal tail may be a common feature of Mlo proteins. PMID- 11904293 TI - Ubiquitous 9-O-acetylation of sialoglycoproteins restricted to the Golgi complex. AB - 9-O-Acetylation of sialic acid is known as a cell type-specific modification of secretory and plasma membrane glycoconjugates of higher vertebrates with important functions in modulating cell-cell recognition. Using a recombinant probe derived from influenza C virus hemagglutinin, we discovered 9-O-acetylated protein in the Golgi complex of various cell lines, most of which did not display 9-O-acetylated sialic acid on the cell surface. All cell lines expressed a sulfated glycoprotein of 50 kDa (sgp50) carrying 9-O-acetylated sialic acids, which was used as a model substrate. Like gp40, the major receptor for influenza C virus of Madin-Darby canine kidney I cells, sgp50 is 9-O-acetylated on O-linked glycans. However, gp40 was not 9-O-acetylated when expressed in Madin-Darby canine kidney II or COS-7 cells. The results demonstrate the existence of two 9-O acetylation machineries for O-glycosylated proteins with distinct substrate specificities. The widespread occurrence of 9-O-acetylated protein in the Golgi furthermore suggests an additional intracellular role for this modification. PMID- 11904294 TI - Phosphorylation by mitogen-activated protein kinase mediates the hypoxia-induced turnover of the TAL1/SCL transcription factor in endothelial cells. AB - The basic helix-loop-helix transcription factor TAL1 (or SCL), originally identified from its involvement by a chromosomal rearrangement in T-cell acute lymphoblastic leukemia, is required for hematopoietic development. TAL1 also has a critical role in embryonic vascular remodeling and is expressed in endothelial cells postnatally, although little is known about its function or regulation in this cell type. We report here that the important proangiogenic stimulus hypoxia stimulates phosphorylation, ubiquitination, and proteasomal breakdown of TAL1 in endothelial cells. Tryptic phosphopeptide mapping and chemical inhibitor studies showed that hypoxia induced the mitogen-activated protein kinase-mediated phosphorylation of a single serine residue, Ser(122), in the protein, and site directed mutagenesis demonstrated that Ser(122) phosphorylation was necessary for hypoxic acceleration of TAL1 turnover in an immortalized murine endothelial cell line. Finally, whereas TAL1 expression was detected in endothelial cells from both large and small vessels, hypoxia-induced TAL1 turnover was observed only in microvascular endothelial cells. Besides their implications for TAL1 function in angiogenic processes, these results demonstrate that a protein kinase(s) important for mitogenic signaling is also utilized in hypoxic endothelial cells to target a transcription factor for destruction. PMID- 11904295 TI - The Escherichia coli cyclic AMP receptor protein forms a 2:2 complex with RNA polymerase holoenzyme, in vitro. AB - Sedimentation equilibrium studies show that the Escherichia coli cyclic AMP receptor protein (CAP) and RNA polymerase holoenzyme associate to form a 2:2 complex in vitro. No complexes of lower stoichiometry (1:1, 2:1, 1:2) were detected over a wide range of CAP and RNA polymerase concentrations, suggesting that the interaction is highly cooperative. The absence of higher stoichiometry complexes, even in the limit of high [protein], suggests that the 2:2 species represents binding saturation for this system. The 2:2 pattern of complex formation is robust. A lower-limit estimate of the formation constant in our standard buffer (40 mm Tris (pH 7.9), 10 mm MgCl(2), 0.1 mm dithiothreitol, 5% glycerol, 100 mm KCl) is 2 x 10(20) m(-3). The qualitative pattern of association is unchanged over the temperature range 4 degrees C < or = T < or = 20 degrees C, by substitution of glutamate for chloride as the dominant anion, or on addition of 20 microm cAMP to the reaction mix. These results limit the possible mechanisms of CAP-polymerase association. In addition, they support the idea that CAP binding may influence the availability of the monomeric form of RNA polymerase that mediates transcription at many promoters. PMID- 11904296 TI - Diphenyleneiodonium triggers the efflux of glutathione from cultured cells. AB - Diphenyleneiodonium (DPI) is a broad-spectrum flavoprotein inhibitor commonly used to inhibit oxidant production by the NADPH oxidase of phagocytic and nonphagocytic cells. A previous study has shown that DPI can sensitize T24 bladder carcinoma cells to Fas-mediated apoptosis. We observed DPI to deplete intracellular reduced glutathione (GSH) in T24 cells and a range of other primary and transformed cell types. The effect was immediate, with 50% loss of intracellular GSH within 2 h of treatment with DPI. The glutathione was quantitatively recovered in the extracellular medium, indicating that efflux was occurring. The loss of GSH was blocked with bromosulfophthalein, an inhibitor of the canalicular GSH transporters. We conclude that DPI induces a dramatic efflux of cellular GSH from T24 cells via a specific transport channel. This provides a potential mechanism for its proapoptotic effect, and it also has important implications for the regulation of glutathione homeostasis in cells. PMID- 11904297 TI - Odontoblast cells immortalized by telomerase produce mineralized dentin-like tissue both in vitro and in vivo. AB - The formation of dentin provides one well accepted paradigm for studying mineralized tissue formation. For the assembly of dentin, several cellular signaling pathways cooperate to provide neural crest-derived mesenchymal cells with positional information. Further, "cross-talk" between signaling pathways from the mesenchymal derived odontoblast cells and the epithelially derived ameloblasts during development is responsible for the formation of functional odontoblasts. These intercellular signals are tightly regulated, both temporally and spatially. When isolated from the developing tooth germ, odontoblasts quickly lose their potential to maintain the odontoblast-specific phenotype. Therefore, generation of an odontoblast cell line would be a valuable reproducible tool for studying the modulatory effects involved in odontoblast differentiation as well as the molecular events involved in mineralized dentin formation. In this study an immortalized odontoblast cell line, which has the required biochemical machinery to produce mineralized tissue in vitro, has been generated. These cells were implanted into animal models to determine their in vivo effects on dentin formation. After implantation, we observed a multistep, programmed cascade of gene expression in the exogenous odontoblasts as the dentin formed de novo. Some of the genes expressed include the dentin matrix proteins 1, 2, and 3, which are extracellular matrix molecules responsible for the ultimate formation of mineralized dentin. The biological response was also examined by histology and radiography and confirmed for mineral deposition by von Kossa staining. Thus, a transformed odontoblast cell line was created with high proliferative capacity that might ultimately be used for the regeneration and repair of dentin in vivo. PMID- 11904298 TI - Genes essential to sodium-dependent bicarbonate transport in cyanobacteria: function and phylogenetic analysis. AB - The cyanobacterium Synechocystis sp. strain PCC 6803 possesses two CO(2) uptake systems and two HCO(3)(-) transporters. We transformed a mutant impaired in CO(2) uptake and in cmpA-D encoding a HCO(3)(-)transporter with a transposon inactivation library, and we recovered mutants unable to take up HCO(3)(-) and grow in low CO(2) at pH 9.0. They are all tagged within slr1512 (designated sbtA). We show that SbtA-mediated transport is induced by low CO(2), requires Na(+), and plays the major role in HCO(3)(-) uptake in Synechocystis. Inactivation of slr1509 (homologous to ntpJ encoding a Na(+)/K(+)-translocating protein) abolished the ability of cells to grow at [Na(+)] higher than 100 mm and severely depressed the activity of the SbtA-mediated HCO(3)(-) transport. We propose that the SbtA-mediated HCO(3)(-) transport is driven by DeltamuNa(+) across the plasma membrane, which is disrupted by inactivating ntpJ. Phylogenetic analyses indicated that two types of sbtA exist in various cyanobacterial strains, all of which possess ntpJ. The sbtA gene is the first one identified as essential to Na(+)-dependent HCO(3)(-) transport in photosynthetic organisms and may play a crucial role in carbon acquisition when CO(2) supply is limited, or in Prochlorococcus strains that do not possess CO(2) uptake systems or Cmp-dependent HCO(3)(-) transport. PMID- 11904299 TI - Residues involved in the mechanism of the bifunctional methylenetetrahydrofolate dehydrogenase-cyclohydrolase: the roles of glutamine 100 and aspartate 125. AB - The human bifunctional dehydrogenase-cyclohydrolase domain catalyzes the interconversion of 5,10-methylene-H(4)folate and 10-formyl-H(4)folate. Although previous structure and mutagenesis studies indicated the importance of lysine 56 in cyclohydrolase catalysis, the role of several surrounding residues had not been explored. In addition to further defining the role of lysine 56, the work presented in this study explores the functions of glutamine 100 and aspartate 125 through the use of site-directed mutagenesis and chemical modification. Mutants at position 100 are inactive with respect to cyclohydrolase activity while preserving significant dehydrogenase levels. We succeeded in producing a K56Q/Q100K double mutant, which has no cyclohydrolase yet retains more than two thirds of wild type dehydrogenase activity. Neither activity is detectable in aspartate 125 mutants with the exception of D125E. The results indicate that the function of glutamine 100 is to activate lysine 56 for cyclohydrolase catalysis and that aspartate 125 is involved in the binding of the H(4)folate substrates. In highlighting the importance of these residues, catalytic mechanisms are proposed for both activities as well as an explanation for the differences in channeling efficiency in the forward and reverse directions. PMID- 11904300 TI - Characterization and regulation of lens-specific calpain Lp82. AB - Eye tissues contain splice variants of muscle-preferred p94 (calpain 3), such as lens-specific Lp82 and Lp85, retina-specific Rt88, and cornea-specific Cn94. The purpose of the present experiment was to analyze the activation and regulation of the best characterized p94 splice variant, Lp82. Recombinant rat Lp82 (rLp82) was expressed using the baculovirus system, purified with Ni-NTA affinity and DEAE ion exchange chromatographies, and characterized by SDS-PAGE, casein zymography, and immunoblotting. After incubation with calcium, rLp82 autolyzed into two major fragments at approximately 60 and 22 kDa. Sequencing of the autolytic fragments showed loss of three amino acids from the N terminus and cleavage near the IS2 region. Also, Lp82 and calpain 2 were found to hydrolyze each other. Calpastatin inhibited calpain 2 activity, but not Lp82. Homology modeling suggested that the lack of inhibition of Lp82 by calpastatin was due to molecular clashes at the unique AX1 region of Lp82. Lp82 also hydrolyzed calpastatin. These results suggested that Lp82 might regulate other calpain activities and cause hydrolysis of substrates such as crystallins during lens cataract formation. PMID- 11904302 TI - The 3'-untranslated region of chloroplast psbA mRNA stabilizes binding of regulatory proteins to the leader of the message. AB - The 5'-leader and 3'-tail of chloroplast mRNAs have been suggested to play a role in posttranscriptional regulation of expression of the message. The regulation is thought to be mediated, at least in part, by regulatory proteins that are encoded by the nuclear genome and targeted to the chloroplast where they interact with chloroplast mRNAs. Previous studies identified high affinity binding of the 5' untranslated region (UTR) of the chloroplast psbA mRNA by Chlamydomonas reinhardtii proteins. Here we tested whether the 3'-UTR of psbA mRNA alone or linked in cis with the 5'-UTR of the mRNA affects the high affinity binding of the message in vitro. We did not detect high affinity binding that is unique to the 3'-UTR. However, we show that the cis-linked 3'-UTR increases the stability of the 5'-UTR binding complex. This effect could provide a means for translational discrimination against mRNAs that are incorrectly processed. PMID- 11904301 TI - Cloning and characterization of a novel mammalian zinc transporter, zinc transporter 5, abundantly expressed in pancreatic beta cells. AB - Intracellular homeostasis for zinc is achieved through the coordinate regulation of specific transporters engaged in zinc influx, efflux, and intracellular compartmentalization. We have identified a novel mammalian zinc transporter, zinc transporter 5 (ZnT-5), by virtue of its similarity to ZRC1, a zinc transporter of Saccharomyces cerevisiae, a member of the cation diffusion facilitator family. Human ZnT-5 (hZnT-5) cDNA encodes a 765-amino acid protein with 15 predicted membrane-spanning domains. hZnT-5 was ubiquitously expressed in all tested human tissues and abundantly expressed in the pancreas. In the human pancreas, hZnT-5 was expressed abundantly in insulin-containing beta cells that contain zinc at the highest level in the body. The hZnT-5 immunoreactivity was found to be associated with secretory granules by electron microscopy. The hZnT-5-derived zinc transport activity was detected using the Golgi-enriched vesicles prepared from hZnT-5-induced HeLa/hZnT-5 cells in which exogenous hZnT-5 expression is inducible by the Tet-on gene regulation system. This activity was dependent on time, temperature, and concentration and was saturable. Moreover, zinc at a high concentration (10 mm) inhibited the growth of yeast expressing hZnT-5. These results suggest that ZnT-5 plays an important role for transporting zinc into secretory granules in pancreatic beta cells. PMID- 11904303 TI - p59Hck isoform induces F-actin reorganization to form protrusions of the plasma membrane in a Cdc42- and Rac-dependent manner. AB - Hck is a protein kinase of the Src family specifically expressed in phagocytes as two isoforms, p59Hck and p61Hck, localized at the plasma membrane and lysosomes, respectively. Their individual involvement in functions ascribed to Hck, phagocytosis, cell migration, and lysosome mobilization, is still unclarified. To investigate the specific role of p59Hck, a constitutively active variant in fusion with green fluorescent protein (p59Hck(ca)) was expressed in HeLa cells. p59Hck(ca) was found at focal adhesion sites and triggered reorganization of the actin cytoskeleton, leading to plasma membrane protrusions where it co-localized with F-actin. Similarly, microinjection of p59Hck(ca) cDNA in J774.A1 macrophages induced membrane protrusions. Whereas kinase activity and membrane association of p59Hck were dispensable for location at focal adhesions, p59Hck-induced membrane protrusions were dependent on kinase activity, plasma membrane association, and Src homology 2 but not Src homology 3 domain and were inhibited by dominant negative forms of Cdc42 or Rac but not by blocking Rho activity. A dominant negative form of p59Hck inhibited the Cdc42- and Rac-dependent FcgammaRIIa mediated phagocytosis. Expression of the Cdc42/Rac-interacting domain of p21 activated kinase in macrophages abolished the p59Hck(ca)-induced morphological changes. Therefore, p59Hck-triggered remodeling of the actin cytoskeleton depends upon the activity of Cdc42 and Rac to promote formation of membrane protrusions necessary for phagocytosis and cell migration. PMID- 11904304 TI - Dynamic interplay between O-glycosylation and O-phosphorylation of nucleocytoplasmic proteins: alternative glycosylation/phosphorylation of THR-58, a known mutational hot spot of c-Myc in lymphomas, is regulated by mitogens. AB - Previously, we reported that c-Myc is glycosylated by O-linked N acetylglucosamine at Thr-58, a known phosphorylation site and a mutational hot spot in lymphomas. In this paper, we describe the production and characterization of two Thr-58 site-specific antibodies and use them to examine the modification of Thr-58 in living cells. One antibody specifically reacts with the Thr-58 glycosylated form of c-Myc, and the other reacts only with unmodified Thr-58 in c Myc. Using these antibodies together with a commercial anti-Thr-58-phosphorylated c-Myc antibody, we simultaneously detected three forms of c-Myc (Thr-58 unmodified, -phosphorylated, and -glycosylated). It has been reported that Thr-58 phosphorylation is dependent on a prior phosphorylation of Ser-62. Mutagenesis of Ser-62 to Ala showed a marked decrease of Thr-58 phosphorylation and a marked increase of Thr-58 glycosylation. Growth inhibition of HL60 cells by serum starvation increases Thr-58 glycosylation and correspondingly decreases its phosphorylation. Serum stimulation has the opposite effect upon the modification status of Thr-58. A candidate kinase responsible for Thr-58 phosphorylation is the glycogen synthase kinase 3 (GSK3). Lithium, a competitive inhibitor of GSK3, decreased Thr-58 phosphorylation and increased its glycosylation. Finally, we show that the Thr-58-phosphorylated form of c-Myc predominantly accumulates in the cytoplasm rather than the nucleus upon inhibition of proteasome activity. These data suggest that hierarchical phosphorylation of Ser-62 and Thr-58 and alternative glycosylation/phosphorylation of Thr-58 together regulate the myriad functions of c-Myc in cells. PMID- 11904305 TI - Identification of two Sp1 phosphorylation sites for p42/p44 mitogen-activated protein kinases: their implication in vascular endothelial growth factor gene transcription. AB - Sp1 regulates activation of many genes implicated in tumor growth and cell cycle progression. We have previously demonstrated its implication in the up-regulation of vascular endothelial growth factor (VEGF) gene transcription following growth factor stimulation of quiescent cells, a situation where p42/p44 mitogen-activate protein kinase (MAPK) activity is dramatically increased. Here we show that p42/p44 MAPK directly phosphorylates Sp1 on threonines 453 and 739 both in vitro and in vivo. Mutation of these sites to alanines decreases by half the MAPK dependent transcriptional activity of Sp1, in the context of the VEGF promoter, in SL2 Drosophila cells devoid of the endogenous Sp1 protein. Moreover, inducible overexpression of the (T453A,T739A) Sp1 double mutant compromises MAPK-driven VEGF mRNA transcription in fibroblasts. These results highlight Sp1 as a key molecular link between elevated activation of the Ras >> p42/p44MAPK signaling pathway and increased VEGF expression, two major steps deregulated in tumor cells. PMID- 11904306 TI - Cyclooxygenase-2 inhibitor therapy for the prevention of esophageal adenocarcinoma in Barrett's esophagus. PMID- 11904307 TI - Androgen deprivation as primary treatment for early prostate cancer: should we "just do something"? PMID- 11904308 TI - Legislators rally for support of revised National Cancer Act. PMID- 11904309 TI - Mammography guidelines in the national spotlight...again. PMID- 11904311 TI - Researchers explore role of gene transfer in tumor growth. PMID- 11904312 TI - Value of prostate-specific antigen: are higher levels meaningful? PMID- 11904313 TI - Intratumoral lymphatic vessels: a case of mistaken identity or malfunction? PMID- 11904314 TI - The effect of selective cyclooxygenase-2 inhibition in Barrett's esophagus epithelium: an in vitro study. AB - BACKGROUND: Individuals with Barrett's esophagus, in which the normal squamous esophageal epithelium is replaced with a columnar mucosa, are at increased risk for esophageal adenocarcinoma. Mucosal injury may be involved in the progression to neoplasia via the synthesis of prostaglandins and other mediators of inflammation. Cyclooxygenase (COX)-2 is the rate-limiting enzyme involved in prostaglandin synthesis. We examined the effect of inhibiting COX-2 activity in Barrett's esophageal cells. METHODS: Primary esophageal epithelial and fibroblast cell cultures were established from endoscopic biopsy specimens from 20 consecutive patients with Barrett's esophagus. COX-2 expression and activity were determined on pooled cell cultures by reverse transcription-polymerase chain reaction and prostaglandin E(2) (PGE(2)) enzyme immunoassay, respectively. Proliferation was measured by Ki-67 staining. PGE(2) levels were determined in supernatants from epithelial cells treated with the selective COX-2 inhibitor NS 398, proinflammatory cytokines (interleukin 1beta and tumor necrosis factor alpha), and conditioned medium from fibroblast cultures (both unstimulated and stimulated with proinflammatory cytokines). RESULTS: Esophageal epithelial cells and fibroblasts expressed COX-2 messenger RNA. Compared with control-treated cells, NS-398 decreased proliferation of Barrett's esophageal epithelial cells by 55% (95% confidence interval = 47.1% to 63.8%; P<.001) and decreased COX-2 activity. The addition of exogenous PGE(2) reversed the antiproliferative effect of NS-398 on Barrett's esophageal epithelial cells. Proinflammatory cytokines did not affect COX-2 activity in esophageal epithelial cells but stimulated COX-2 activity in fibroblasts. However, conditioned medium from unstimulated and stimulated fibroblasts increased COX-2 activity in esophageal epithelial cells. CONCLUSION: COX-2 is functionally active in Barrett's esophagus because treatment with the COX-2 inhibitor hinders proliferation of Barrett's esophageal epithelial cells in culture, but proliferation is restored by treatment with prostaglandin. These results raise the possibility that inhibition of COX-2 may have chemopreventive potential for Barrett's esophagus. PMID- 11904315 TI - Quality of life following localized prostate cancer treated initially with androgen deprivation therapy or no therapy. AB - BACKGROUND: Many men diagnosed with clinically localized prostate cancer are initially treated conservatively, receiving neither surgery nor radiotherapy for the first year. Treatment patterns and quality-of-life outcomes have not been previously reported for a population-based sample of such men. METHODS: A population-based random sample of men (n = 661) from six geographic regions who had been newly diagnosed with clinically localized prostate cancer from 1994 through 1995 were followed for up to 1 year. Eligible subjects received neither surgery nor radiotherapy within 1 year of initial diagnosis. We assessed disease specific and generic quality-of-life outcomes in men receiving androgen deprivation therapy (ADT) compared with men receiving no therapy. All statistical tests were two-sided. RESULTS: Two hundred and forty-five study patients received ADT and the remaining 416 patients received no therapy. Approximately two thirds of the patients (n = 159) receiving ADT had either baseline Gleason scores greater than six or serum prostate-specific antigen values above 20 ng/mL. Among men who were sexually potent before diagnosis (ADT = 88 patients; no therapy = 223 patients), 80% of those on ADT reported being impotent after 1 year compared with 30% of those receiving no treatment (P < .001). Patients receiving ADT reported more physical discomfort 1 year after diagnosis than did men who had received no therapy. However, patients receiving ADT, compared with those receiving no therapy, were more likely to be satisfied with their treatment decision (56% pleased versus 45.3%; P =.001). Patients on ADT also experienced a statistically significant decline in vitality, but not in physical function, after adjustment for the confounding factors (P =.05). CONCLUSION: ADT is a commonly used primary therapy for clinically localized prostate cancer. Therefore, men considering ADT as an initial treatment should be aware that sexual function and some aspects of physical well-being are likely to be affected in the first year following this treatment. PMID- 11904316 TI - Calcium intake and risk of colon cancer in women and men. AB - BACKGROUND: Calcium has been hypothesized to reduce the risk of colon cancer, and in a recent randomized trial, calcium supplementation was associated with reduction in the risk of recurrent colorectal adenomas. We examined the association between calcium intake and colon cancer risk in two prospective cohorts, the Nurses' Health Study (NHS) and the Health Professionals Follow-up Study (HPFS). METHODS: Our study population included 87 998 women in NHS and 47 344 men in HPFS who, at baseline (1980 for NHS and 1986 for HPFS), completed a food frequency questionnaire and provided information on medical history and lifestyle factors. Dietary information was updated at least every 4 years. During the follow-up period (1980 to May 31, 1996 for the NHS cohort; 1986 to January 31, 1996 for the HPFS cohort), 626 and 399 colon cancer cases were identified in women and men, respectively. Pooled logistic regression was used to estimate relative risks (RRs), and all statistical tests were two-sided. RESULTS: In women and men considered together, we found an inverse association between higher total calcium intake (>1250 mg/day versus < or =500 mg/day) and distal colon cancer (women: multivariate RR = 0.73, 95% confidence interval [CI] = 0.41 to 1.27; men: RR = 0.58, 95% CI = 0.32 to 1.05; pooled RR = 0.65, 95% CI = 0.43 to 0.98). No such association was found for proximal colon cancer (women: RR = 1.28, 95% CI = 0.75 to 2.16; men: RR = 0.92, 95% CI = 0.45 to 1.87; pooled RR = 1.14, 95% CI = 0.72 to 1.81). The incremental benefit of additional calcium intake beyond approximately 700 mg/day appeared to be minimal. CONCLUSIONS: Higher calcium intake is associated with a reduced risk of distal colon cancer. The observed risk pattern was consistent with a threshold effect, suggesting that calcium intake beyond moderate levels may not be associated with a further risk reduction. Future investigations on this association should concentrate on specific cancer subsites and on the dose-response relationship. PMID- 11904317 TI - p16(Ink4a) in melanocyte senescence and differentiation. AB - BACKGROUND: The Ink4a-Arf tumor suppressor locus encodes two growth inhibitors, p16 and Arf, both of which are also implicated as effectors in cellular senescence. Because human germline defects in the INK4A-ARF locus are associated with familial melanoma, melanocytes may have unusual INK4A-ARF functions or controls of cell senescence. Because senescence is believed to be an anticancer mechanism, we investigated the role of Ink4a-Arf and its individual components in melanocyte senescence. METHODS: Melanocytes were cultured from littermate mice with zero, one, or two functional copies of the Ink4a-Arf locus. Senescence was evaluated by cumulative population doubling curves and by the assessment of acidic beta-galactosidase (an indicator of senescence) expression. Pigmentation and cell size were evaluated by spectrophotometry and microscopy. p16 and Arf expression in primary and spontaneously immortalized melanocyte or melanocyte precursor cell lines were evaluated by immunoblotting. Retroviral vectors containing normal p16 and Arf complementary DNAs were used to restore expression of these genes in Ink4a-Arf(-/-) melanocytes. RESULTS: Wild-type melanocytes (i.e., Ink4a-Arf(+/+)) senesced within 4-5 weeks of culture. Ink4a-Arf(-/-) melanocytes did not senesce and readily became immortal. Ink4a-Arf(+/-) melanocytes showed defective senescence. Senescent Ink4a-Arf(+/+) melanocytes were heavily pigmented, but Ink4a-Arf(+/-) and Ink4a-Arf(-/-) melanocytes were less pigmented. All of six spontaneously immortalized melanocyte or melanocyte precursor lines from Ink4a-Arf(+/+) mice lacked p16 protein expression, although most retained Arf protein expression. After restoration of p16 but not Arf expression, Ink4a-Arf(-/-) melanocytes stopped growing, became highly melanized, and expressed acidic beta-galactosidase. By contrast, restoration of Arf but not p16 expression led to cell death without evidence of senescence. CONCLUSION: Normal mouse melanocyte senescence and associated pigmentation require both copies of Ink4a-Arf and appear to depend more on p16 than on Arf function. Mutations of the INK4A-ARF locus may favor tumorigenesis from melanocytes by impairing senescence, cell differentiation, and (where ARF is disrupted) cell death. PMID- 11904318 TI - Association of a common polymorphism in the human GH1 gene with colorectal neoplasia. AB - BACKGROUND: Growth hormone (GH) may be associated with the development of colorectal tumors directly and/or indirectly via an increased plasma level of insulin-like growth factor-I (IGF-I), which has been associated with colorectal cancer risk. Because a T-to-A polymorphism in the human GH1 gene at position 1663 is putatively associated with lower levels of GH and IGF-I, we investigated the relationship of this polymorphism to the risk of colorectal neoplasia. METHODS: We analyzed data from two case-control studies conducted in Hawaii: a population based study of 535 case patients with colorectal adenocarcinoma and 650 control subjects and a sigmoidoscopy screening-based study with 139 case patients with adenoma and 202 control subjects. All subjects were tested for the GH1 polymorphism. Logistic regression was used to adjust for known risk factors. Plasma IGF-I and IGF binding protein-3 (IGFBP-3) levels were measured in a subset of 293 subjects in the adenoma study (135 case patients and 158 control subjects). All statistical tests were two-sided. RESULTS: Adjusted odds ratios (ORs) for colorectal cancer associated with T/T, T/A, and A/A genotypes were 1.00, 0.75 (95% confidence interval [CI] = 0.58 to 0.99), and 0.62 (95% CI = 0.43 to 0.90), respectively (P(trend) =.006). Adjusted ORs for adenoma were 1.00, 0.76 (95% CI = 0.46 to 1.24), and 0.62 (95% CI = 0.31 to 1.22), respectively (P(trend) =.17). Data from both studies consistently showed that the A allele was associated with a lower risk of colorectal neoplasia than the T allele, although the association with adenoma was not statistically significant. These associations were consistently suggested in Caucasians and Native Hawaiians but not in Japanese. The ratio of plasma IGF-I/IGFBP-3 was lower in individuals with the A allele than in individuals with the T allele (P =.01). CONCLUSION: The human T1663A GH1 gene polymorphism, which may confer lower levels of GH and IGF I, appears to be associated with a decreased risk of colorectal cancer. PMID- 11904319 TI - Re: familial multiple myeloma: a family study and review of the literature. PMID- 11904320 TI - Re: familial multiple myeloma: a family study and review of the literature. PMID- 11904324 TI - The Mozena Classification System and treatment algorithm for ingrown hallux nails. AB - Infected ingrown toenails raise the question of how much nail should be removed and what amount of nail fold reduction should occur. Often, the ungual labia folds are found to be hypertrophic, forcing the nail to push into the flesh and start a foreign body reaction. A simplified approach to this problem is proposed on the basis of the measurement of 100 normal nail folds and 25 infected nail folds. The results of this study show that the treatment goal should be to achieve an ungual labia fold of less than 3 mm, concluding that there is a correlation between the depth of the ungual labia fold and the severity of the infected ingrown toenail. PMID- 11904323 TI - Symptomatic reversal of peripheral neuropathy in patients with diabetes. AB - Forty-nine consecutive subjects with established diabetic peripheral neuropathy were treated with monochromatic near-infrared photo energy (MIRE) to determine if there was an improvement of sensation. Loss of protective sensation characterized by Semmes-Weinstein monofilament values of 4.56 and above was present in 100% of subjects (range, 4.56 to 6.45), and 42 subjects (86%) had Semmes-Weinstein values of 5.07 or higher. The ability to discriminate between hot and cold sensation was absent (54%) or impaired (46%) in both groups prior to the initiation of MIRE treatment. On the basis of Semmes-Weinstein monofilament values, 48 subjects (98%) exhibited improved sensation after 6 treatments, and all subjects had improved sensation after 12 treatments. Therefore, MIRE may be a safe, drug-free, noninvasive treatment for the consistent and predictable improvement of sensation in diabetic patients with peripheral neuropathy of the feet. PMID- 11904325 TI - Clinical and dynamic range-of-motion techniques in subjects with and without diabetes mellitus. AB - Thirty subjects with type 1 diabetes, 30 subjects with type 2 diabetes, and 30 age- and sex-matched controls were evaluated through clinical goniometry and two dimensional motion analysis systems to determine the dynamic and static range of motion of the knee, ankle, and hallux joints. The purpose of this study was to determine if the knee and ankle joints of patients with diabetes mellitus are affected by limited joint mobility syndrome. The study results support previous medical literature showing significant reduction of range of motion of the hallux in subjects with type 1 diabetes. Significant differences were found between the range of motion of male and female subjects in all lower-limb joints for both subject groups with diabetes compared to the control group, and male subjects in all groups recorded less range of motion than female subjects. PMID- 11904326 TI - A comparative study of lactic acid 10% and ammonium lactate 12% lotion in the treatment of foot xerosis. AB - Xerotic skin is a pattern of reaction to a variety of disorders that have abnormalities of desquamation in common. This double-blind, randomized clinical trial investigated the effect of Lactinol (Pedinol Pharmaceuticals, Farmingdale, New York) versus Lac-Hydrin 12% (Bristol-Myers Squibb, Princeton, New Jersey) lotion in mild to moderate foot xerosis. Clinical assessment of xerosis was performed at baseline visit, and the designated sites were evaluated at 2 and 4 weeks after treatment began. Of the 53 patients enrolled, 18 were excluded from analysis. Although both treatment groups had significantly improved xerosis scores after 2 and 4 weeks of treatment, no statistically significant difference was observed. Of the 44% of patients who did express a preference, 72% preferred Lactinol, which may account for the 20% increase in its overall use in the study. PMID- 11904327 TI - Pedal gangrene secondary to disseminated intravascular coagulation with gastric carcinoma. AB - Consumptive coagulopathy resulting in a disseminated intravascular coagulation is most often seen in infectious diseases and hematologic malignancies. Solid tumors may be associated with disseminated intravascular coagulation that results in gangrene of the upper extremity. A case report of lower-extremity gangrene as the pathology for gastric carcinoma is presented. The need for a multidisciplinary approach to this clinical presentation is noted. PMID- 11904328 TI - Congenital absence of the tibial sesamoid. AB - Congenital absence of the tibial sesamoid of the hallux is an infrequent occurrence. The authors present a case of congenital absence of the tibial sesamoid and a review of the literature regarding the clinical significance of this anomaly. PMID- 11904329 TI - Capillary hemangioma of the foot. AB - Capillary hemangiomas are benign, vascular lesions of skin and mucous membranes that often occur in infancy and childhood. Capillary hemangiomas are most commonly found in the head and neck region. Capillary hemangiomas that occur in adults and on the lower extremities are uncommon. A clinical case involving surgical treatment of the lesion on an adult foot is presented. PMID- 11904331 TI - 'Horses for Courses': the differences between quantitative and qualitative approaches to research. AB - Some clinicians may feel dissociated from, and intimidated by the ever-increasing emphasis on research. However, with an understanding of some of the basic principles and key terms, research can feel less daunting. It is the aim of this article, the second in a series of three focusing on understanding research, to introduce clinicians to the different approaches to research, to improve understanding of what the approaches mean, and to highlight when a particular approach may be appropriate. Furthermore, the article will provide an explanation of some of the common terms used within clinical research. This should aid the clinician in applying good, simple, scientific principles to evaluating clinical research evidence. PMID- 11904332 TI - Council on Podiatric Medical Education: eighty-second annual report, 2001. PMID- 11904334 TI - Neurofibromin and NF1 gene analysis in composite pheochromocytoma and tumors associated with von Recklinghausen's disease. AB - Composite tumor of pheochromocytoma and neuroblastoma, or ganglioneuroma, or ganglioneuroblastoma (composite pheochromocytoma), also known as mixed neuroendocrine and neural tumor, are sometimes combined with neurofibromatosis type 1 (NF1). To better understand the relationship between NF1 and composite pheochromocytoma, an immunohistochemical study using anti-neuro-fibromin that is an NF1 gene product and DNA sequence of NF1 Exon 31 were carried out in five cases of composite pheochromocytoma and in various tumors from five patients with NF1. Neurofibromin was not expressed in Schwann cells and sustentacular cells of composite pheochromocytomas and was very weakly or negatively expressed in neurofibroma of NF1 patients. However, it was strongly expressed in ganglionic cells and pheochromocytoma cells of the composite pheochromocytomas and also in mucosal ganglioneuromas, a gangliocytic paraganglioma, and in pheochromocytomas from the patients with NF1. Although there was no mutation in NF1 Exon 31, it could not be ruled out that there were mutations in other sites of the NF1 gene. Neurofibromin insufficiency may induce abnormal proliferation of Schwann cells in composite pheochromocytomas as well as in neurofibromatosis. PMID- 11904335 TI - Hepatic adenomas: analysis of sex steroid receptor status and the Wnt signaling pathway. AB - Hepatic adenomas are strongly linked to excess hormonal exposure, but little else is known about their pathogenesis. The Wnt signaling pathway, which is activated in both hepatocellular carcinomas and hepatoblastomas, has not been studied in hepatic adenomas. Fifteen hepatic adenomas were studied by immunohistochemistry for estrogen, progesterone, and androgen receptors (ER, PR, AR, respectively) and correlated with the results of immunostaining for beta-catenin. Direct sequencing was performed to look for mutations in key genes involved in the Wnt signaling pathway: Exon 3 of beta-catenin encompassing the glycogen synthase kinase 3beta (GSK-3beta) phosphorylation region and the mutational cluster region of the adenomatosis polyposis coli protein (APC). Analysis for loss of heterozygosity (LOH) at chromosome 5q was also performed. Immunostaining for both ER and PR was present in 11/15 (73%) adenomas, and staining with one hormone receptor was positively associated with staining for the other receptor. AR positivity was present in 3/15 cases. Nuclear accumulation of beta-catenin was present in 7/15 (46%) of adenomas, indicating activation of the Wnt signaling pathway. However, no beta-catenin mutations, no APC mutations in the mutational cluster region, and no 5q LOH were detected. Two APC polymorphisms of unknown significance were seen. No clear association between beta-catenin nuclear accumulation and hormone receptor positivity was discerned. Activation of the Wnt signaling pathway appears to be important in a subset of hepatic adenomas but does not result from common beta-catenin or APC mutations and does not appear to be directly linked to hormonal receptor status. PMID- 11904336 TI - Antineutrophil cytoplasmic autoantibody in the absence of Wegener's granulomatosis or microscopic polyangiitis: implications for the surgical pathologist. AB - Antineutrophil cytoplasmic antibodies (ANCA) are useful serologic markers for the diagnosis and management of patients with Wegener's granulomatosis (WG) and microscopic polyangiitis (MPA). However, problems in diagnosis and classification may occur when patients with other disorders develop ANCA. A 7-year review (1993 1999) disclosed 247 patients whose sera tested positively for ANCA by an indirect immunofluorescence method: 166 patients for cytoplasmic-ANCA (C-ANCA) and 81 patients for perinuclear-ANCA (P-ANCA) Twenty-seven patients had active pulmonary disease and underwent open-lung biopsy or transbronchial biopsy. Eight patients (30%) had a disease other than WG or MPA, and their clinical, pathological, and serological findings were reviewed. The patients, all women, ranged in age from 28 to 77 years (median, 37 y). Dyspnea (n = 6), cough (n = 6), chest pain (n = 2), and/or hemoptysis (n = 2) were present. The duration of symptoms lasted from 3 weeks to 6 years (median, 6 mo). ANCA titers were C-ANCA (n = 4; range, 1:40 1280) or P-ANCA (n = 4; range, 1:40-640). The lung biopsies disclosed nonspecific interstitial pneumonia (n = 4), bronchiolitis obliterans organizing pneumonia (n = 1), diffuse alveolar damage (n = 1), organizing diffuse alveolar hemorrhage without capillaritis (n = 1), and necrotic granuloma (n = 1). No cases showed characteristic histology for WG or MPA. The final diagnoses were various connective tissue disorders (n = 5), chronic hypersensitivity pneumonia (n = 1), postinfectious bronchitis/bronchiectasis (n = 1), and ulcerative colitis-related lung disease (n = 1). Surgical pathologists should be aware that significantly elevated ANCA titers may be associated with diverse forms of pulmonary disease. ANCA positivity alone, in the absence of appropriate clinical or pathologic findings, should not be used to substantiate a diagnosis of WG or MPA. PMID- 11904337 TI - Mutational analysis of the von hippel lindau gene in clear cell renal carcinomas from tuberous sclerosis complex patients. AB - Tuberous sclerosis complex (TSC) is an autosomal-dominant disorder characterized by seizures, mental retardation, autism, and tumors of multiple organs. Renal disease in TSC includes angiomyolipomas, cysts, and renal cell carcinomas. It is known that somatic mutations in the von Hippel Lindau (VHL) tumor suppressor gene occur in most clear cell renal carcinomas. To determine whether TSC-associated clear cell carcinomas also contain VHL mutations, we analyzed six tumors for loss of heterozygosity in the VHL gene region of chromosome 3p and for mutations in the VHL gene. Four of the patients were women between the ages of 34 and 68 years, and two were males under the age of 21 years. The loss of heterozygosity analysis was performed using polymorphic microsatellite markers, and the mutational analysis was performed using direct sequencing. Chromosome 3p loss of heterozygosity was not detected, and no VHL mutations were identified. These findings suggest that mutations in the TSC1 and TSC2 genes lead to clear cell renal carcinogenesis via an alternate pathway not involving VHL mutations. PMID- 11904338 TI - Analysis of octamer-binding transcription factors Oct2 and Oct1 and their coactivator BOB.1/OBF.1 in lymphomas. AB - Oct1 and Oct2 are transcription factors of the POU homeo-domain family that bind to the Ig gene octamer sites, regulating B-cell-specific genes. The function of these transcription factors is dependent on the activity of B-cell-restricted coactivators such as BOB.1/OBF.1. Independent studies of the expression of these proteins in non-Hodgkin's lymphoma have been restricted to single markers, and most lack data concerning immunohistochemical expression. Thus, we have investigated the expression of Oct1, Oct2, and BOB.1/OBF.1 in human reactive lymphoid tissue and in a series of 140 Hodgkin and non-Hodgkin's lymphomas. None of these proteins was found to be restricted to B cells, although only B cells expressed high levels of all three markers. Additionally, germinal center B cells showed stronger Oct2 and BOB.1/OBF.1 staining. Consequently, most B-cell lymphomas showed reactivity for all three antibodies. Oct2 expression was significantly higher in germinal center-derived lymphomas, although other B-cell lymphomas also displayed a high level of Oct2 expression. Although T-cell lymphomas and Hodgkin's lymphomas expressed some of these proteins, they commonly exhibited less reactivity than B-cell lymphomas. Despite not being entirely cell specific, the strong nuclear expression of Oct2 and BOB.1/OBF.1 by germinal center- derived lymphomas makes these antibodies a potentially useful tool in lymphoma diagnosis. PMID- 11904339 TI - A brief history of head and neck pathology. AB - Surgical pathology had its beginnings in the late 1800s. A biopsy that gained much attention was from the larynx of Crown Prince Frederick in 1887. The tissue was seen by Rudolph Virchow and the clinical management of the Prince eventuated in a highly publicized furor. During the first half of the twentieth century, numerous entities in the head and neck were described by dozens of pathologists worldwide. The information was scattered in clinical journals for radiotherapists, general surgeons, and otolaryngologists. The first book on ear, nose, and throat pathology did not appear until 1947 and by 1956 two atlases were available. The book was "Histopathology of the Ear, Nose and Throat" by Eggston and Wolff (1947), and the atlases were the first Armed Forces Institute of Pathology (AFIP) fascicle on salivary gland tumors by Foote and Frazell (1954) and "An Atlas of Otolaryngic Pathology" by Ash and Raum (1956). Clinicopathologic studies accelerated in the 1960s as laryngeal conservation therapy evolved and radiation therapy became more sophisticated. The years 1968 and 1974 mark major events for the emergence of Head and Neck Pathology into a clear-cut discipline. In 1968, Vincent J. Hyams was appointed Director of Otolaryngic Pathology at the AFIP, and 1974 was the publication date of "Tumors of the Head and Neck" by John G. Batsakis. The past 25 years have been filled with hundreds of articles on new entities and the application of fresh technology to old entities. Specialized therapeutic approaches have demanded greater diagnostic precision. This paper touches on a few representative aspects in the history of Head and Neck Pathology during the past 130 years. PMID- 11904340 TI - Squamous cell carcinoma of the upper aerodigestive tract: precursors and problematic variants. PMID- 11904342 TI - Neuroectodermal neoplasms of the head and neck with emphasis on neuroendocrine carcinomas. AB - Tumors exhibiting neuroectodermal differentiation occur throughout the body, and the diverse tissues of the head and neck give rise to a wide assortment of these neoplasms. Neuroectodermal neoplasms may be divided into lesions showing primarily epithelial differentiation (Group I, neuroendocrine carcinomas) and a more diverse group (Group II) of nonepithelial neoplasms. This article reviews these neuroectodermal tumors of the head and neck with emphasis on the neuroendocrine carcinomas and their nomenclature. The author believes that with regard to Group I tumors, the older terminology of carcinoid, atypical carcinoid, and small cell carcinoma should be replaced by subclassifications of well differentiated, moderately differentiated, and poorly differentiated neuroendocrine carcinoma. The latter category should be further subdivided into small cell and large cell variants. Neuroendocrine carcinomas, particularly the moderately differentiated subtype, are often underdiagnosed in the head and neck region. In the larynx, these tumors are the most common form of nonsquamous carcinoma. Poorly differentiated neuroendocrine carcinoma of small cell type is most common in the salivary glands but can occur elsewhere in the region. The large cell subtype of poorly differentiated neuroendocrine carcinoma has not been well documented in this region. However, the most likely candidate for this tumor category is the so-called sinonasal undifferentiated carcinoma. Group II tumors discussed include olfactory neuroblastoma, malignant melanoma, and Ewing's sarcoma. In addition, differential diagnostic problems related to Group I and II tumors are reviewed in detail. This article reviews and updates our understanding of neuroectodermal neoplasms arising in the head and neck. The focus is on tumors that exclusively involve this region or show a strong predilection to occur here. PMID- 11904341 TI - Lymphoid lesions of the head and neck: a model of lymphocyte homing and lymphomagenesis. AB - Lymphoid lesions of the head and neck mainly affect the nasopharynx, nasal and paranasal sinuses, and salivary glands. These three compartments each are affected by a different spectrum of lymphoid malignancies and can serve as model for mechanisms of lymphomagenesis. The type of lymphoma seen reflects the underlying biology and function of the particular site involved. The nasopharynx and Waldeyer's ring are functionally similar to the mucosal associated lymphoid tissue (MALT) of the gastrointestinal tract and are most commonly affected by B cell lymphomas, with mantle cell lymphoma being a relatively frequent subtype. The most prevalent lymphoid lesion of the salivary gland is lymphoepithelial sialadenitis, associated with Sjogren's syndrome. Lymphoepithelial sialadenitis is a condition in which MALT is acquired in a site not normally containing lymphoid tissue. Patients with Sjogren's syndrome are at increased risk to develop B-cell lymphomas, most commonly MALT lymphomas. The nasal and paranasal sinuses are the prototypical site for the development of extranodal natural killer (NK) /T-cell lymphoma, nasal type. This condition must be distinguished from other conditions causing the clinical picture of lethal midline granuloma, including Wegener's granulomatosis and infectious disorders. Lymphomatoid granulomatosis is common in the lung but is rarely seen in the midline facial structures. PMID- 11904343 TI - Schneiderian papillomas and nonsalivary glandular neoplasms of the head and neck. AB - Schneiderian papillomas and nonsalivary glandular neoplasms of the head and neck continue to be a source of confusion for both the clinician and pathologist. An update on these lesions is provided. PMID- 11904344 TI - Salivary gland neoplasia: a review for the practicing pathologist. PMID- 11904345 TI - Distinctive soft tissue tumors of the head and neck. AB - Among soft tissue tumors as a whole, those in the head and neck region are relatively uncommon, and the proportion of all soft tissue sarcomas that arise in this region is 250 microM). The more physiologically relevant conditions of either peroxynitrite infusion (1 microM/min) or de novo formation by simultaneous, equimolar generation of nitric oxide (NO) and superoxide (e.g., 3-morpholinosydnonimine; NONOates plus xanthine oxidase/hypoxanthine, menadione, or mitomycin C) were examined. Despite robust oxidation of dihydrorhodamine under each of these conditions, fluorescence decrease of both purified and intracellular GFP was not evident regardless of carbon dioxide presence, suggesting that oxidation and nitration are not necessarily coupled. Alternatively, both extra- and intracellular GFP fluorescence was exquisitely sensitive to nitration produced by heme peroxidase/hydrogen peroxide-catalyzed oxidation of nitrite. Formation of nitrogen dioxide (NO(2)) during the reaction between NO and the nitroxide 2 phenyl-4,4,5,5-tetramethylimidazole-1-oxyl 3-oxide indicated that NO(2) can enter cells and alter peptide function through tyrosyl nitration. Taken together, these findings exemplified that heme-peroxidase-catalyzed formation of NO(2) may play a pivotal role in inflammatory and chronic disease settings while calling into question the significance of nitration by peroxynitrite. PMID- 11904415 TI - Elongator is a histone H3 and H4 acetyltransferase important for normal histone acetylation levels in vivo. AB - The elongating, hyperphosphorylated form of RNA polymerase II is associated with the Elongator complex, which has the histone acetyltransferase (HAT) Elp3 as a subunit. Here we show that, in contrast to the isolated Elp3 subunit, the activity of intact Elongator complex is directed specifically toward the amino terminal tails of histone H3 and H4, and that Elongator can acetylate both core histones and nucleosomal substrates. The predominant acetylation sites are lysine 14 of histone H3 and lysine-8 of histone H4. The three smallest Elongator subunits--Elp4, Elp5, and Elp6--are required for HAT activity, and Elongator binds to both naked and nucleosomal DNA. By using chromatin immunoprecipitation, we show that the levels of multiply acetylated histone H3 and H4 in chromatin are decreased in vivo in yeast cells lacking ELP3. PMID- 11904417 TI - How the folding rate constant of simple, single-domain proteins depends on the number of native contacts. AB - Experiments have shown that the folding rate constants of two dozen structurally unrelated, small, single-domain proteins can be expressed in terms of one quantity (the contact order) that depends exclusively on the topology of the folded state. Such dependence is unique in chemical kinetics. Here we investigate its physical origin and derive the approximate formula ln(k) = ln(N) + a + bN, were N is the number of contacts in the folded state, and a and b are constants whose physical meaning is understood. This formula fits well the experimentally determined folding rate constants of the 24 proteins, with single values for a and b. PMID- 11904416 TI - Identification and characterization of a human DNA glycosylase for repair of modified bases in oxidatively damaged DNA. AB - 8-oxoguanine (8-oxoG), ring-opened purines (formamidopyrimidines or Fapys), and other oxidized DNA base lesions generated by reactive oxygen species are often mutagenic and toxic, and have been implicated in the etiology of many diseases, including cancer, and in aging. Repair of these lesions in all organisms occurs primarily via the DNA base excision repair pathway, initiated with their excision by DNA glycosylase/AP lyases, which are of two classes. One class utilizes an internal Lys residue as the active site nucleophile, and includes Escherichia coli Nth and both known mammalian DNA glycosylase/AP lyases, namely, OGG1 and NTH1. E. coli MutM and its paralog Nei, which comprise the second class, use N terminal Pro as the active site. Here, we report the presence of two human orthologs of E. coli mutM nei genes in the human genome database, and characterize one of their products. Based on the substrate preference, we have named it NEH1 (Nei homolog). The 44-kDa, wild-type recombinant NEH1, purified to homogeneity from E. coli, excises Fapys from damaged DNA, and oxidized pyrimidines and 8-oxoG from oligodeoxynucleotides. Inactivation of the enzyme because of either deletion of N-terminal Pro or Histag fusion at the N terminus supports the role of N-terminal Pro as its active site. The tissue-specific levels of NEH1 and OGG1 mRNAs are distinct, and S phase-specific increase in NEH1 at both RNA and protein levels suggests that NEH1 is involved in replication associated repair of oxidized bases. PMID- 11904418 TI - Mutation of the myosin converter domain alters cross-bridge elasticity. AB - Elastic distortion of a structural element of the actomyosin complex is fundamental to the ability of myosin to generate motile forces. An elastic element allows strain to develop within the actomyosin complex (cross-bridge) before movement. Relief of this strain then drives filament sliding, or more generally, movement of a cargo. Even with the known crystal structure of the myosin head, however, the structural element of the actomyosin complex in which elastic distortion occurs remained unclear. To assign functional relevance to various structural elements of the myosin head, e.g., to identify the elastic element within the cross-bridge, we studied mechanical properties of muscle fibers from patients with familial hypertrophic cardiomyopathy with point mutations in the head domain of the beta-myosin heavy chain. We found that the Arg-719 --> Trp (Arg719Trp) mutation, which is located in the converter domain of the myosin head fragment, causes an increase in force generation and fiber stiffness under isometric conditions by 48-59%. Under rigor and relaxing conditions, fiber stiffness was 45-47% higher than in control fibers. Yet, kinetics of active cross-bridge cycling were unchanged. These findings, especially the increase in fiber stiffness under rigor conditions, indicate that cross-bridges with the Arg719Trp mutation are more resistant to elastic distortion. The data presented here strongly suggest that the converter domain that forms the junction between the catalytic and the light-chain-binding domain of the myosin head is not only essential for elastic distortion of the cross bridge, but that the main elastic distortion may even occur within the converter domain itself. PMID- 11904419 TI - Mechanical force generation by G proteins. AB - GTP-hydrolyzing G proteins are molecular switches that play a critical role in cell signaling processes. Here we use molecular dynamics simulations to show that Ras, a monomeric G protein, can generate mechanical force upon hydrolysis. The generated force levels are comparable to those produced by ATP-hydrolyzing motor proteins, consistent with the structural similarities of the catalytic region of motor proteins and G proteins. The force transduction mechanism is based on an irreversible structural change, produced by the hydrolysis, which triggers thermal switching between force-generating substates through changes in the configurational space of the protein. PMID- 11904420 TI - Fold prediction of helical proteins using torsion angle dynamics and predicted restraints. AB - We describe a procedure for predicting the tertiary folds of alpha-helical proteins from their primary sequences. The central component of the procedure is a method for predicting interhelical contacts that is based on a helix-packing model. Instead of predicting the individual contacts, our method attempts to identify the entire patch of contacts that involve residues regularly spaced in the sequences. We use this component to glue together two powerful existing methods: a secondary structure prediction program, whose output serves as the input to the contact prediction algorithm, and the tortion angle dynamics program, which uses the predicted tertiary contacts and secondary structural states to assemble three-dimensional structures. In the final step, the procedure uses the initial set of simulated structures to refine the predicted contacts for a new round of structure calculation. When tested against 24 small to medium sized proteins representing a wide range of helical folds, the completely automated procedure is able to generate native-like models within a limited number of trials consistently. PMID- 11904421 TI - Haploinsufficiency of mTR results in defects in telomere elongation. AB - Telomeres are usually maintained about an equilibrium length, and the set point for this equilibrium differs between species and between strains of a given species. To examine the requirement for telomerase in mediating establishment of a new telomere length equilibrium, we generated interspecies crosses with telomerase mTR knockout mice. In crosses between C57BL/6J (B6) and either of two unrelated mouse species, CAST/Ei and SPRET/Ei, telomerase mediated establishment of a new telomere length equilibrium in wild-type mTR(+/+) mice. This new equilibrium was characterized by elongation of the short telomeres of CAST/Ei or SPRET/Ei origin. In contrast, mTR(-/-) offspring of interspecies crosses failed to elongate telomeres. Unexpectedly, haploinsufficiency was observed in mTR(+/-) heterozygous interspecies mice, which had an impaired ability to elongate short SPRET/Ei or CAST/Ei telomeres to the new equilibrium set point that was achieved in wild-type mTR(+/+) mice. These results demonstrate that elongation of telomeres to a new telomere set point requires telomerase and indicate that telomerase RNA may be limiting in vivo. PMID- 11904422 TI - Preferential maintenance of critically short telomeres in mammalian cells heterozygous for mTert. AB - Prolonged growth of murine embryonic stem (ES) cells lacking the telomerase reverse transcriptase, mTert, results in a loss of telomere DNA and an increased incidence of end-to-end fusions and aneuploidy. Furthermore, loss of only one copy of mTert also results in telomere shortening intermediate between wild-type (wt) and mTert-null ES cells [Liu, Y., Snow, B. E., Hande, M. P., Yeung, D., Erdmann, N. J., Wakeham, A., Itie, A., Siderovski, D. P., Lansdorp, P. M., Robinson, M. O. & Harrington, L. (2000) Curr. Biol. 10, 1459-1462]. Unexpectedly, although average telomere length in mTert(+/-) ES cells declined to a similar level as mTert-null ES cells, mTert(+/-) ES cell lines retained a minimal telomeric DNA signal at all chromosome ends. Consequently, no end-to-end fusions and genome instability were observed in the latest passages of mTert(+/-) ES cell lines. These data uncover a functional distinction between the dosage-dependent function of telomerase in average telomere-length maintenance and the selective maintenance of critically short telomeres in cells heterozygous for mTert. In normal and tumor cells, we suggest that telomerase activity insufficient to maintain a given average telomere length may, nonetheless, provide a protective advantage from end-to-end fusion and genome instability. PMID- 11904423 TI - Neutrophils lacking phosphoinositide 3-kinase gamma show loss of directionality during N-formyl-Met-Leu-Phe-induced chemotaxis. AB - Confocal imaging and time-lapsed videomicroscopy were used to study the directionality, motility, rate of cell movement, and morphologies of phosphoinositide 3-kinase gamma (PI3K)gamma(-/-) neutrophils undergoing chemotaxis in Zigmond chambers containing N-formyl-Met-Leu-Phe gradients. Most of the PI3Kgamma(-/-) neutrophils failed to translocate up the chemotactic gradient. A partial reduction in cell motility and abnormal morphologies were also observed. In the wild-type neutrophils, the pleckstrin homology domain-containing protein kinase B (AKT) and F-actin colocalize to the leading edge of polarized neutrophils oriented toward the gradient, which was not observed in PI3Kgamma(-/ ) neutrophils. In PI3Kgamma(-/-) neutrophils, AKT staining consistently failed to perfectly overlap with the F-actin. This failure was observed as an F-actin filled region of 2.3 +/- 0.5 microm between AKT and the cell membrane. These data suggest that PI3Kgamma regulates neutrophil chemotaxis primarily by controlling the direction of cell migration and the intracellular colocalization of AKT and F actin to the leading edge. PMID- 11904424 TI - Integrins regulate the apoptotic response to DNA damage through modulation of p53. AB - p53 mediates apoptosis of cells after DNA damage including tumor cells after radiation or chemotherapy. Survival of isolated cancer cells after therapy leads to recurrence of therapy-resistant tumors. We now show that for some melanoma, sarcoma, or fibroblastic cell types that survive without integrin-mediated adhesion, apoptosis in response to DNA damage requires cell adhesion. This effect correlates with rapid changes in levels of p14/p19 Arf and its downstream component, p53. Killing of nonadherent cells is increased by treatment with antiintegrin antibodies or by increasing levels of p53 or Arf. Consistent with low p53 levels, suspended cells show chromosomal instability after irradiation. Thus, loss of normal adhesion in susceptible tumor cells during genotoxic stress may play a role in therapy resistance and promote survival of cells with aberrant DNA. PMID- 11904426 TI - Predicting the spatial dynamics of rabies epidemics on heterogeneous landscapes. AB - Often as an epidemic spreads, the leading front is irregular, reflecting spatial variation in local transmission rates. We developed a methodology for quantifying spatial variation in rates of disease spread across heterogeneous landscapes. Based on data for epidemic raccoon rabies in Connecticut, we developed a stochastic spatial model of rabies spread through the state's 169 townships. We quantified spatial variation in transmission rates associated with human demography and key habitat features. We found that large rivers act as semipermeable barriers, leading to a 7-fold reduction in the local rates of propagation. By combining the spatial distribution of major rivers with long distance dispersal we were able to account for the observed irregular pattern of disease spread across the state without recourse to direct assessment of host pathogen populations. PMID- 11904425 TI - Immortalization of a primate bipotent epithelial liver stem cell. AB - Liver regeneration after partial hepatectomy results primarily from the simple division of mature hepatocytes. However, during embryonic and fetal development or in circumstances under which postnatal hepatocytes are injured, organ regeneration is believed to occur from a compartment of epithelial liver stem or progenitor cells with biliary and hepatocytic bipotentiality. The ability to identify, isolate, and transplant epithelial liver stem cells from fetal liver would greatly facilitate the treatment of hepatic diseases currently requiring orthotopic liver transplantation. Here we report the identification and immortalization by retrovirus-mediated transfer of the simian virus 40 large T antigen gene of primate fetal epithelial liver cells with a dual hepatocytic biliary phenotype. These cells grow indefinitely in vitro and express the liver epithelial cell markers cytokeratins 8/18, the hepatocyte-specific markers albumin and alpha-fetoprotein, and the biliary-specific markers cytokeratins 7 and 19. Bipotentiality of gene expression was confirmed by clonal analysis initiated from single cells. Endogenous telomerase also is expressed constitutively. After orthotopic transplantation via the portal vein, approximately 50% of the injected cells integrated into the liver parenchyma of athymic mice without tumorigenicity. Three weeks after transplantation, cells having seeded in the liver parenchyma expressed both albumin and alpha fetoprotein but had lost expression of cytokeratin 19. These results provide strong evidence for the existence of a bipotent epithelial liver stem cell in nonhuman primates. This unlimited source of donor cells also should enable the establishment of a model of allogenic liver cell transplantation in a large animal closely related to humans and shed light on important questions related to liver organogenesis and differentiation. PMID- 11904427 TI - Rapid prehistoric extinction of iguanas and birds in Polynesia. AB - The Tongoleleka archaeological site on Lifuka Island, Kingdom of Tonga, is a rich accumulation of pottery, marine mollusks, and nonhuman bones that represents first human contact on a small island in Remote Oceania approximately 2,850 years ago. The lower strata contain decorated Lapita-style pottery and bones of an extinct iguana (Brachylophus undescribed sp.) and numerous species of extinct birds. The upper strata instead feature Polynesian Plainware pottery and bones of extant species of vertebrates. A stratigraphic series of 20 accelerator-mass spectrometer radiocarbon dates on individual bones of the iguana, an extinct megapode (Megapodius alimentum), and the non-native chicken (Gallus gallus) suggests that anthropogenic loss of the first two species and introduction of the latter occurred on Lifuka within a time interval too short (a century or less) to be resolved by radiometric dating. The geologically instantaneous prehistoric collapse of Lifuka's vertebrate community contrasts with the much longer periods of faunal depletion on some other islands, thus showing that the elapse time between human arrival and major extinction events was highly variable on oceanic islands as well as on continents. PMID- 11904428 TI - Metabolic efficiency and amino acid composition in the proteomes of Escherichia coli and Bacillus subtilis. AB - Biosynthesis of an Escherichia coli cell, with organic compounds as sources of energy and carbon, requires approximately 20 to 60 billion high-energy phosphate bonds [Stouthamer, A. H. (1973) Antonie van Leeuwenhoek 39, 545-565]. A substantial fraction of this energy budget is devoted to biosynthesis of amino acids, the building blocks of proteins. The fueling reactions of central metabolism provide precursor metabolites for synthesis of the 20 amino acids incorporated into proteins. Thus, synthesis of an amino acid entails a dual cost: energy is lost by diverting chemical intermediates from fueling reactions and additional energy is required to convert precursor metabolites to amino acids. Among amino acids, costs of synthesis vary from 12 to 74 high-energy phosphate bonds per molecule. The energetic advantage to encoding a less costly amino acid in a highly expressed gene can be greater than 0.025% of the total energy budget. Here, we provide evidence that amino acid composition in the proteomes of E. coli and Bacillus subtilis reflects the action of natural selection to enhance metabolic efficiency. We employ synonymous codon usage bias as a measure of translation rates and show increases in the abundance of less energetically costly amino acids in highly expressed proteins. PMID- 11904429 TI - Enhancer action in trans is permitted throughout the Drosophila genome. AB - Interactions between paired homologous genes can lead to changes in gene expression. Such trans-regulatory effects exemplify transvection and are displayed by many genes in Drosophila, in which homologous chromosomes are paired somatically. Transvection involving the yellow cuticle pigmentation gene can occur by at least two mechanisms, one involving the trans-action of enhancers on a paired promoter and a second involving pairing-mediated bypass of a chromatin insulator. A system was developed to evaluate whether the action of the yellow enhancers in trans could be reconstituted outside of the natural near telomeric location of the yellow gene. To this end, transgenic flies were generated that carried a yellow gene modified by the inclusion of strategically placed recognition sites for the Cre and FLP recombinases. Independent action of the recombinases produced a pair of derivative alleles, one enhancerless and the other promoterless, at each transgene location. Transvection between the derivatives was assessed by the degree of interallelic complementation. Complementation was observed at all eight sites tested. These studies demonstrate that yellow transvection can occur at multiple genomic locations and indicate that the Drosophila genome generally is permissive to enhancer action in trans. PMID- 11904430 TI - The Dun1 checkpoint kinase phosphorylates and regulates the ribonucleotide reductase inhibitor Sml1. AB - Cell cycle checkpoints are evolutionarily conserved surveillance systems that protect genomic stability and prevent oncogenesis in mammals. One important target of checkpoint control is ribonucleotide reductase (RNR), which catalyzes the rate-limiting step in dNTP and DNA synthesis. In both yeast and humans, RNR is transcriptionally induced after DNA damage via Mec1/Rad53 (yeast) and ATM/CHK2 (human) checkpoint pathways. In addition, yeast checkpoint proteins Mec1 and Rad53 also regulate the RNR inhibitor Sml1. After DNA damage or at S phase, Mec1 and Rad53 control the phosphorylation and concomitant degradation of Sml1 protein. This new layer of control contributes to the increased dNTP production likely necessary for DNA repair and replication; however, the molecular mechanism is unclear. Here we show that Dun1, a downstream kinase of Mec1/Rad53, genetically and physically interacts with Sml1 in vivo. The absence of Dun1 activity leads to the accumulation of Sml1 protein at S phase and after DNA damage. As a result, dun1Delta strains need more time to finish DNA replication, are defective in mitochondrial DNA propagation, and are sensitive to DNA-damaging agents. Moreover, phospho-Sml1 is absent or dramatically reduced in dun1Delta cells. Finally, Dun1 can phosphorylate Sml1 in vitro. These results suggest that Dun1 kinase function is the last step required in the Mec1/Rad53 cascade to remove Sml1 during S phase and after DNA damage. PMID- 11904431 TI - A screen for dominant mutations applied to components in the Drosophila EGF-R pathway. AB - The Drosophila epidermal growth factor receptor (EGF-R) controls many critical cell fate choices throughout development. Several proteins collaborate to promote localized EGF-R activation, such as Star and Rhomboid (Rho), which act sequentially to ensure the maturation and processing of inactive membrane-bound EGF ligands. To gain insights into the mechanisms underlying Rho and Star function, we developed a mutagenesis scheme to isolate novel overexpression activity (NOVA) alleles. In the case of rho, we isolated a dominant neomorphic allele, which interferes with Notch signaling, as well as a dominant-negative allele, which produces RNA interference-like flip-back transcripts that reduce endogenous rho expression. We also obtained dominant-negative and neomorphic Star mutations, which have phenotypes similar to those of rho NOVA alleles, as well as dominant-negative Egf-r alleles. The isolation of dominant alleles in several different genes suggests that NOVA mutagenesis should be widely applicable and emerge as an effective tool for generating dominant mutations in genes of unknown function. PMID- 11904433 TI - Negative regulation of Lck by Cbl ubiquitin ligase. AB - The Cbl-family ubiquitin ligases function as negative regulators of activated receptor tyrosine kinases by facilitating their ubiquitination and subsequent targeting to lysosomes. Cbl associates with the lymphoid-restricted nonreceptor tyrosine kinase Lck, but the functional relevance of this interaction remains unknown. Here, we demonstrate that T cell receptor and CD4 coligation on human T cells results in enhanced association between Cbl and Lck, together with Lck ubiquitination and degradation. A Cbl(-/-) T cell line showed a marked deficiency in Lck ubiquitination and increased levels of kinase-active Lck. Coexpression in 293T cells demonstrated that Lck kinase activity and Cbl ubiquitin ligase activity were essential for Lck ubiquitination and negative regulation of Lck dependent serum response element-luciferase reporter activity. The Lck SH3 domain was pivotal for Cbl-Lck association and Cbl-mediated Lck degradation, with a smaller role for interactions mediated by the Cbl tyrosine kinase-binding domain. Finally, analysis of a ZAP-70-deficient T cell line revealed that Cbl inhibited Lck-dependent mitogen-activated protein kinase activation, and an intact Cbl RING finger domain was required for this functional effect. Our results demonstrate a direct, ubiquitination-dependent, negative regulatory role of Cbl for Lck in T cells, independent of Cbl-mediated regulation of ZAP-70. PMID- 11904432 TI - DNA-dependent protein kinase suppresses double-strand break-induced and spontaneous homologous recombination. AB - DNA-dependent protein kinase (DNA-PK), composed of Ku70, Ku80, and the catalytic subunit (DNA-PKcs), is involved in repairing double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ). Certain proteins involved in NHEJ are also involved in DSB repair by homologous recombination (HR). To test the effects of DNA-PKcs on DSB-induced HR, we integrated neo direct repeat HR substrates carrying the I-SceI recognition sequence into DNA-PKcs-defective Chinese hamster ovary (V3) cells. The DNA-PKcs defect was complemented with a human DNA-PKcs cDNA. DSB-induced HR frequencies were 1.5- to 3-fold lower with DNA-PKcs complementation. In complemented and uncomplemented strains, all products arose by gene conversion without associated crossover, and average conversion tract lengths were similar. Suppression of DSB-induced HR in complemented cells probably reflects restoration of NHEJ, consistent with competition between HR and NHEJ during DSB repair. Interestingly, spontaneous HR rates were 1.6- to >3.5 fold lower with DNA-PKcs complementation. DNA-PKcs may suppress spontaneous HR through NHEJ of spontaneous DSBs, perhaps at stalled or blocked replication forks. Because replication protein A (RPA) is involved in both replication and HR, and is phosphorylated by DNA-PKcs, it is possible that the suppression of spontaneous HR by DNA-PKcs reflects regulation of replication-dependent HR by DNA PKcs, perhaps by means of phosphorylation of RPA. PMID- 11904434 TI - Prions in skeletal muscle. AB - Considerable evidence argues that consumption of beef products from cattle infected with bovine spongiform encephalopathy (BSE) prions causes new variant Creutzfeldt-Jakob disease. In an effort to prevent new variant Creutzfeldt-Jakob disease, certain "specified offals," including neural and lymphatic tissues, thought to contain high titers of prions have been excluded from foods destined for human consumption [Phillips, N. A., Bridgeman, J. & Ferguson-Smith, M. (2000) in The BSE Inquiry (Stationery Office, London), Vol. 6, pp. 413-451]. Here we report that mouse skeletal muscle can propagate prions and accumulate substantial titers of these pathogens. We found both high prion titers and the disease causing isoform of the prion protein (PrP(Sc)) in the skeletal muscle of wild type mice inoculated with either the Me7 or Rocky Mountain Laboratory strain of murine prions. Particular muscles accumulated distinct levels of PrP(Sc), with the highest levels observed in muscle from the hind limb. To determine whether prions are produced or merely accumulate intramuscularly, we established transgenic mice expressing either mouse or Syrian hamster PrP exclusively in muscle. Inoculating these mice intramuscularly with prions resulted in the formation of high titers of nascent prions in muscle. In contrast, inoculating mice in which PrP expression was targeted to hepatocytes resulted in low prion titers. Our data demonstrate that factors in addition to the amount of PrP expressed determine the tropism of prions for certain tissues. That some muscles are intrinsically capable of accumulating substantial titers of prions is of particular concern. Because significant dietary exposure to prions might occur through the consumption of meat, even if it is largely free of neural and lymphatic tissue, a comprehensive effort to map the distribution of prions in the muscle of infected livestock is needed. Furthermore, muscle may provide a readily biopsied tissue from which to diagnose prion disease in asymptomatic animals and even humans. PMID- 11904435 TI - Profound defects in pancreatic beta-cell function in mice with combined heterozygous mutations in Pdx-1, Hnf-1alpha, and Hnf-3beta. AB - Defects in pancreatic beta-cell function contribute to the development of type 2 diabetes, a polygenic disease that is characterized by insulin resistance and compromised insulin secretion. Hepatocyte nuclear factors (HNFs) -1alpha, -3beta, -4alpha, and Pdx-1 contribute in the complex transcriptional circuits within the pancreas that are involved in beta-cell development and function. In mice, a heterozygous mutation in Pdx-1 alone, but not Hnf-1alpha(+/-), Hnf-3beta(+/-), or Hnf-4alpha(+/-), causes impaired glucose-stimulated insulin secretion in mice. To investigate the possible functional relationships between these transcription factors on beta-cell activity in vivo, we generated mice with the following combined heterozygous mutations: Pdx-1(+/-)/Hnf-1alpha(+/-), Pdx-1(+/-)/Hnf 3beta(+/-), Pdx-1(+/-)/Hnf-4alpha(+/-), Hnf-1alpha(+/-)/Hnf-4alpha(+/-), and Hnf 3beta(+/-)/Hnf-4alpha(+/-). The greatest loss in function was in combined heterozygous null alleles of Pdx-1 and Hnf-1alpha (Pdx-1(+/-)/Hnf-1alpha(+/-)), or Pdx-1 and Hnf-3beta (Pdx-1(+/-)/Hnf-3beta(+/-)). Both double mutants develop progressively impaired glucose tolerance and acquire a compromised first- and second-phase insulin secretion profile in response to glucose compared with Pdx 1(+/-) mice alone. The loss in beta-cell function in Pdx-1(+/-)/Hnf-3beta(+/-) mice was associated with decreased expression of Nkx-6.1, glucokinase (Gck), aldolase B (aldo-B), and insulin, whereas Nkx2.2, Nkx-6.1, Glut-2, Gck, aldo-B, the liver isoform of pyruvate kinase, and insulin expression was reduced in Pdx 1(+/-)/Hnf-1alpha(+/-) mice. The islet cell architecture was also abnormal in Pdx 1(+/-)/Hnf-3beta(+/-) and Pdx-1(+/-)/Hnf-1alpha(+/-) mice, with glucagon expressing cells scattered throughout the islet, a defect that may be connected to decreased E-cadherin expression. Our data suggest that functional interactions between key islet regulatory factors play an important role in maintaining islet architecture and beta-cell function. These studies also established polygenic mouse models for investigating the mechanisms contributing to beta-cell dysfunction in diabetes. PMID- 11904436 TI - The lipofuscin component A2E selectively inhibits phagolysosomal degradation of photoreceptor phospholipid by the retinal pigment epithelium. AB - Daily phagocytosis of spent photoreceptor outer segments is a critical maintenance function performed by the retinal pigment epithelium (RPE) to preserve vision. Aging RPE accumulates lipofuscin, which includes N-retinylidene N-retinylethanolamine (A2E) as the major autofluorescent component. We studied the effect of physiological levels of A2E in RPE cultures on their ability to phagocytose outer segments. A2E localized to lysosomes in cultured RPE as well as in human RPE in situ. A2E-loaded RPE cells in culture bound and internalized identical numbers of outer segments as control RPE indicating that A2E does not alter early steps of phagocytosis. A2E-loaded RPE degraded outer segment proteins efficiently but, strikingly, failed to completely digest phospholipids within 24 h. Because of the circadian rhythm of RPE phagocytosis in the eye, a delay in lipid degradation would likely result in a build up of undigested material in RPE that could contribute to the development of age-related macular degeneration. PMID- 11904437 TI - Immunization of Aotus monkeys with a functional domain of the Plasmodium falciparum variant antigen induces protection against a lethal parasite line. AB - Immunity to Plasmodium falciparum in African children has been correlated with antibodies to the P. falciparum erythrocyte membrane protein 1 (PfEMP1) variant gene family expressed on the surface of infected red cells. We immunized Aotus monkeys with a subregion of the Malayan Camp variant antigen (MCvar1) that mediates adhesion to the host receptor CD36 on the endothelial surface and present data that PfEMP1 is an important target for vaccine development. The immunization induced a high level of protection against the homologous strain. Protection correlated with the titer of agglutinating antibodies and occurred despite the expression of variant copies of the gene during recurrent waves of parasitemia. A second challenge with a different P. falciparum strain, to which there was no agglutinating activity, showed no protection but boosted the immune response to this region during the infection. The level of protection and the evidence of boosting during infection encourage further exploration of this concept for malaria vaccine development. PMID- 11904438 TI - G protein-coupled receptor kinase 4 gene variants in human essential hypertension. AB - Essential hypertension has a heritability as high as 30-50%, but its genetic cause(s) has not been determined despite intensive investigation. The renal dopaminergic system exerts a pivotal role in maintaining fluid and electrolyte balance and participates in the pathogenesis of genetic hypertension. In genetic hypertension, the ability of dopamine and D(1)-like agonists to increase urinary sodium excretion is impaired. A defective coupling between the D(1) dopamine receptor and the G protein/effector enzyme complex in the proximal tubule of the kidney is the cause of the impaired renal dopaminergic action in genetic rodent and human essential hypertension. We now report that, in human essential hypertension, single nucleotide polymorphisms of a G protein-coupled receptor kinase, GRK4gamma, increase G protein-coupled receptor kinase (GRK) activity and cause the serine phosphorylation and uncoupling of the D(1) receptor from its G protein/effector enzyme complex in the renal proximal tubule and in transfected Chinese hamster ovary cells. Moreover, expressing GRK4gammaA142V but not the wild type gene in transgenic mice produces hypertension and impairs the diuretic and natriuretic but not the hypotensive effects of D(1)-like agonist stimulation. These findings provide a mechanism for the D(1) receptor coupling defect in the kidney and may explain the inability of the kidney to properly excrete sodium in genetic hypertension. PMID- 11904439 TI - The identification of E2F1-specific target genes. AB - The E2F family of transcriptional regulators consists of six different members. Analysis of E2F-regulated promoters by using cultured cells suggests that E2Fs may have redundant functions. However, animal studies have shown that loss of individual E2Fs can have distinct biological consequences. Such seemingly conflicting results could be due to a difference in E2F-mediated regulation in cell culture vs. animals. Alternatively, there may be genes that are specifically regulated by an individual E2F which have not yet been identified. To investigate this possibility further, we have analyzed gene expression in E2F1 nullizygous mice. We found that loss of E2F1 did not cause changes in expression of known E2F target genes, suggesting that perhaps E2F1-specific promoters are distinct from known E2F target promoters. Therefore, we used oligonucleotide microarrays to identify mRNAs whose expression is altered on loss of E2F1. We demonstrate by chromatin immunoprecipitation that several of the promoters that drive expression of the deregulated mRNAs selectively recruit E2F1, but not other E2Fs, and this recruitment is via an element distinct from a consensus E2F binding site. To our knowledge, these are as yet undocumented examples of promoters being occupied in asynchronously growing cells by a single E2F family member. Interestingly, the E2F1-specific target genes that we identified encode proteins having functions quite different from the function of known E2F target genes. Thus, whereas E2F1 may share redundant functions in cell growth control with other E2F family members, it may also play an important biological role distinct from the other E2Fs. PMID- 11904440 TI - Deficient Smad7 expression: a putative molecular defect in scleroderma. AB - Scleroderma is a chronic systemic disease that leads to fibrosis of affected organs. Transforming growth factor (TGF) beta has been implicated in the pathogenesis of scleroderma. Smad proteins are signaling transducers downstream from TGF-beta receptors. Three families of Smads have been identified: (i) receptor-regulated Smad2 and -3 (R-Smads); (ii) common partner Smad4 (Co-Smad); and (iii) inhibitory Smad6 and -7 (I-Smads, part of a negative feedback loop). We have investigated the signaling components for the TGF-beta pathway and TGF-beta activity in scleroderma lesions in vivo and in scleroderma fibroblasts in vitro. Basal level and TGF-beta-inducible expression of Smad7 are selectively decreased, whereas Smad3 expression is increased both in scleroderma skin and in explanted scleroderma fibroblasts in culture. TGF-beta signaling events, including phosphorylation of Smad2 and -3, and transcription of the PAI-1 gene are increased in scleroderma fibroblasts, relative to normal fibroblasts. In vitro adenoviral gene transfer with Smad7 restores normal TGF-beta signaling in scleroderma fibroblasts. These results suggest that alterations in the Smad pathway, including marked Smad7 deficiency and Smad3 up-regulation, may be responsible for TGF-beta hyperresponsiveness observed in scleroderma. PMID- 11904442 TI - Chronic intracellular infection of alfalfa nodules by Sinorhizobium meliloti requires correct lipopolysaccharide core. AB - Our analyses of lipopolysaccharide mutants of Sinorhizobium meliloti offer insights into how this bacterium establishes the chronic intracellular infection of plant cells that is necessary for its nitrogen-fixing symbiosis with alfalfa. Derivatives of S. meliloti strain Rm1021 carrying an lpsB mutation are capable of colonizing curled root hairs and forming infection threads in alfalfa in a manner similar to a wild-type strain. However, developmental abnormalities occur in the bacterium and the plant at the stage when the bacteria invade the plant nodule cells. Loss-of-function lpsB mutations, which eliminate a protein of the glycosyltransferase I family, cause striking changes in the carbohydrate core of the lipopolysaccharide, including the absence of uronic acids and a 40-fold relative increase in xylose. We also found that lpsB mutants were sensitive to the cationic peptides melittin, polymyxin B, and poly-l-lysine, in a manner that paralleled that of Brucella abortus lipopolysaccharide mutants. Sensitivity to components of the plant's innate immune system may be part of the reason that this mutant is unable to properly sustain a chronic infection within the cells of its host-plant alfalfa. PMID- 11904441 TI - The quantity of nitric oxide released by macrophages regulates Chlamydia-induced disease. AB - Intracellular bacteria of the genus Chlamydia cause numerous typically chronic diseases, frequently with debilitating sequelae. Genetic determinants of disease susceptibility after infection with Chlamydia bacteria are unknown. C57BL/6 mice develop severe pneumonia and poor immunity against Chlamydia after moderate respiratory infection whereas BALB/c mice are protected from disease and develop vigorous Th1 immunity. Here we show that infected C57BL/6 macrophages release more NO synthesized by NO synthase 2 (NOS2) than BALB/c macrophages and have lower mRNA concentrations of arginase II, a competitor of NOS2 for the common substrate, l-arginine. Reduction, but not elimination, of NO production by incomplete inhibition of NOS2 abolishes susceptibility of C57BL/6 mice to Chlamydia-induced disease. Thus, the quantity of NO released by infected macrophages is the effector mechanism that regulates between pathogenic and protective responses to chlamydial infection, and genes controlling NO production determine susceptibility to chlamydial disease. PMID- 11904443 TI - Dominant-interfering forms of MEF2 generated by caspase cleavage contribute to NMDA-induced neuronal apoptosis. AB - Myocyte enhancer factor-2 (MEF2) transcription factors are activated by p38 mitogen-activated protein kinase during neuronal and myogenic differentiation. Recent work has shown that stimulation of this pathway is antiapoptotic during development but proapoptotic in mature neurons exposed to excitotoxic or other stress. We now report that excitotoxic (N-methyl-D-aspartate) insults to mature cerebrocortical neurons activate caspase-3, -7, in turn cleaving MEF2A, C, and D isoforms. MEF2 cleavage fragments containing a truncated transactivation domain but preserved DNA-binding domain block MEF2 transcriptional activity via dominant interference. Transfection of constitutively active MEF2 (MEF2C-CA) rescues MEF2 transcriptional activity after N-methyl-D-aspartate insult and prevents neuronal apoptosis. Conversely, dominant-interfering MEF2 abrogates neuroprotection by MEF2C-CA. These results define a pathway to excitotoxic neuronal stress/apoptosis via caspase-catalyzed cleavage of MEF2. Additionally, we show that similar MEF2 cleavage fragments are generated in vivo during focal stroke damage. Hence, this pathway appears to have pathophysiological relevance in vivo. PMID- 11904444 TI - Induction of behavioral associative memory by stimulation of the nucleus basalis. AB - The nucleus basalis (NB) has been implicated in memory formation indirectly, by lesions, pharmacological manipulations, and neural correlates of learning. Prior findings imply that engagement of the NB during learning promotes memory storage. We directly tested this NB-memory hypothesis by determining whether stimulation of the NB induces behavioral associative memory. Rats were trained either with paired tone (6 kHz) and NB stimulation or with the two stimuli unpaired. We later determined the specificity of cardiac and respiratory behavioral responses to the training tone and several other acoustic frequencies. Paired subjects exhibited frequency generalization gradients with a peak of 6 kHz for both cardiac and respiratory behavior. Unpaired subjects exhibited no generalization gradient. The development of such specific, associative behavioral responses indicates that tone paired with NB stimulation induced behavioral associative memory. The discovery of memory induction by direct activation of the NB supports the NB memory hypothesis and provides a potentially powerful way to control and investigate neural mechanisms of memory. PMID- 11904445 TI - The vitelliform macular dystrophy protein defines a new family of chloride channels. AB - Vitelliform macular dystrophy (VMD/Best disease; MIM*153700) is an early-onset autosomal dominant disorder in which accumulation of lipofuscin-like material within and beneath the retinal pigment epithelium is associated with a progressive loss of central vision. Bestrophin, the protein product of the VMD gene, has four predicted transmembrane domains. There are multiple bestrophin homologues in the human, Drosophila, and Caenorhabditis elegans genomes, but no function has previously been ascribed to these proteins, and they show no detectable homology to other proteins of known function. Using heterologous expression, we show here that human, Drosophila, and C. elegans bestrophins form oligomeric chloride channels, and that human bestrophin is sensitive to intracellular calcium. Each of 15 missense mutations asscociated with VMD greatly reduces or abolishes the membrane current. Four of these mutant bestrophins were coexpressed with the wild type and each dominantly inhibited the wild-type membrane current, consistent with the dominant nature of the disease. These experiments establish the existence of a new chloride channel family and VMD as a channelopathy. PMID- 11904446 TI - Updating of the visual representation in monkey striate and extrastriate cortex during saccades. AB - Neurons in the lateral intraparietal area, frontal eye field, and superior colliculus exhibit a pattern of activity known as remapping. When a salient visual stimulus is presented shortly before a saccade, the representation of that stimulus is updated, or remapped, at the time of the eye movement. This updating is presumably based on a corollary discharge of the eye movement command. To investigate whether visual areas also exhibit remapping, we recorded from single neurons in extrastriate and striate cortex while monkeys performed a saccade task. Around the time of the saccade, a visual stimulus was flashed either at the location occupied by the neuron's receptive field (RF) before the saccade (old RF) or at the location occupied by it after the saccade (new RF). More than half (52%) of V3A neurons responded to a stimulus flashed in the new RF even though the stimulus had already disappeared before the saccade. These neurons responded to a trace of the flashed stimulus brought into the RF by the saccade. In 16% of V3A neurons, remapped activity began even before saccade onset. Remapping also was observed at earlier stages of the visual hierarchy, including in areas V3 and V2. At these earlier stages, the proportion of neurons that exhibited remapping decreased, and the latency of remapped activity increased relative to saccade onset. Remapping was very rare in striate cortex. These results indicate that extrastriate visual areas are involved in the process of remapping. PMID- 11904447 TI - Frequency-dependent synchrony in locus ceruleus: role of electrotonic coupling. AB - Electrotonic coupling synchronizes the spontaneous firing of locus ceruleus (LC) neurons in the neonatal rat brain, whereas in adults, synchronous activity is rare. This report examines the role of action potential frequency on synchronous activity in the adult LC. Decreasing the firing frequency in slices from adult animals facilitated the appearance of subthreshold oscillations and increased the correlation of the membrane potential between pairs of neurons. Conversely, increasing the firing frequency decreased the amplitude and synchrony of the oscillations among pairs. The frequency-dependent synchrony was not observed in slices from neonatal rats, where synchrony was observed at all frequencies, suggesting a developmental change in the properties of the LC network. A mathematical model confirmed that a reduction of the coupling strength among a pair of coupled neurons could generate frequency-dependent synchrony. In slices from adult animals, the combination of electrotonic coupling and firing frequency are the key elements that regulate synchronous firing in this nucleus. PMID- 11904448 TI - Wild-type and mutated presenilins 2 trigger p53-dependent apoptosis and down regulate presenilin 1 expression in HEK293 human cells and in murine neurons. AB - Presenilins 1 and 2 are two homologous proteins that, when mutated, account for most early onset Alzheimer's disease. Several lines of evidence suggest that, among various functions, presenilins could modulate cell apoptotic responses. Here we establish that the overexpression of presenilin 2 (PS2) and its mutated form Asn-141-Ile-PS2 alters the viability of human embryonic kidney (HEK)293 cells as established by combined trypan blue exclusion, sodium 3'-[1-(phenylamino carbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene sulfonic acid hydrate assay, and propidium iodide incorporation FACS analyses. The two parent proteins increase the acetyl-DEVD-al-sensitive caspase-3-like activity in both HEK293 cells and Telencephalon specific murine neurons, modulate Bax and bcl-2 expressions, and enhance cytochrome C translocation into the cytosol. We show that overexpression of both wild-type and mutated PS2 increases p53-like immunoreactivity and transcriptional activity. We also establish that wild-type- and mutated PS2-induced caspase activation is reduced by p53 antisense approach and by pifithrin-alpha, a chemical inhibitor of p53. Furthermore, mouse fibroblasts in which the PS2 gene has been knocked out exhibited strongly reduced p53-transcriptional activity. Finally, we establish that the overexpression of both wild-type and mutated PS2 is accompanied by a drastic reduction of endogenous presenilin 1 (PS1) expression. Interestingly, pifithrin-alpha diminished endogenous PS2 immunoreactivity, whereas the inhibitor increases PS1 expression. Altogether, our data demonstrate that wild-type and familial Alzheimer's disease-linked PS2 trigger apoptosis and down-regulate PS1 expression through p53-dependent mechanisms. PMID- 11904449 TI - Testosterone attenuates expression of vascular cell adhesion molecule-1 by conversion to estradiol by aromatase in endothelial cells: implications in atherosclerosis. AB - We previously reported that testosterone attenuated atherogenesis in LDLR(-/-) male mice, and that this effect of testosterone was most likely caused by its conversion to estradiol. Estradiol inhibits vascular cell adhesion molecule-1 (VCAM-1) expression, and expression of VCAM-1 is one of the early events in atherogenesis. We assessed the cellular mechanism(s) involved by which testosterone attenuates atherogenesis. We evaluated whether testosterone inhibited TNFalpha-induced VCAM-1 expression via its conversion to estradiol by the enzyme aromatase in human umbilical vein endothelial cells (HUVEC). Aromatase mRNA was dedected by reverse transcription-PCR in these cells. Testosterone (30 nM-1 microM) attenuated VCAM-1 mRNA expression in a concentration-dependent manner. The non aromatizable androgen, dihydrotestosterone, had no effect on VCAM 1 mRNA expression. Testosterone was less effective in attenuating VCAM-1 expression in the presence of anastrozole, an inhibitor of aromatase, indicating that testosterone inhibited VCAM-1 via conversion to estradiol. Estradiol also attenuated VCAM-1 mRNA expression, but this action was not abolished in the presence of anastrozole, indicating that anastrozole itself did not modulate VCAM 1 mRNA expression. The effect of testosterone on VCAM-1 mRNA expression was inhibited in the presence of the estrogen receptor antagonist, ICI-182780. Testosterone also attenuated TNFalpha-induced VCAM-1 protein expression, and this attenuation was abolished in the presence of anastrozole. In conclusion, testosterone inhibited VCAM-1 mRNA and protein expression in HUVEC by its conversion to estradiol via the enzyme aromatase present in the endothelial cells. Results from our study may help explain the mechanism by which testosterone may have beneficial effects on the cardiovascular system. PMID- 11904451 TI - Short day lengths augment stress-induced leukocyte trafficking and stress-induced enhancement of skin immune function. AB - Environmental conditions influence the onset and severity of infection and disease. Stressful conditions during winter may weaken immune function and further compromise survival by means of hypothermia, starvation, or shock. To test the hypothesis that animals may use photoperiod to anticipate the onset of seasonal stressors and adjust immune function, we evaluated glucocorticoids and the distribution of blood leukocytes in Siberian hamsters (Phodopus sungorus) exposed to long day lengths (i.e., summer) or short day (SD) lengths (i.e., winter) at baseline and during acute stress. We also investigated the influence of photoperiod and acute stress on a delayed-type hypersensitivity response in the skin. SDs increased glucocorticoid concentrations and the absolute number of circulating blood leukocytes, lymphocytes, T cells, and natural killer cells at baseline in hamsters. During stressful challenges, it appears beneficial for immune cells to exit the blood and move to primary immune defense areas such as the skin, in preparation for potential injury or infection. Acute (2 h) restraint stress induced trafficking of lymphocytes and monocytes out of the blood. This trafficking occurred more rapidly in SDs compared to long days. Baseline delayed type hypersensitivity responses were enhanced during SDs; this effect was augmented by acute stress and likely reflected more rapid redistribution of leukocytes out of the blood and into the skin. These results suggest that photoperiod may provide a useful cue by which stressors in the environment may be anticipated to adjust the repertoire of available immune cells and increase survival likelihood. PMID- 11904450 TI - Akt-dependent phosphorylation of endothelial nitric-oxide synthase mediates penile erection. AB - In the penis, nitric oxide (NO) can be formed by both neuronal NO synthase and endothelial NOS (eNOS). eNOS is activated by viscous drag/shear stress in blood vessels to produce NO continuously, a process mediated by the phosphatidylinositol 3-kinase (PI3kinase)/Akt pathway. Here we show that PI3 kinase/Akt physiologically mediates erection. Both electrical stimulation of the cavernous nerve and direct intracavernosal injection of the vasorelaxant drug papaverine cause rapid increases in phosphorylated (activated) Akt and eNOS. Phosphorylation is diminished by wortmannin and LY294002, inhibitors of PI3 kinase, the upstream activator of Akt. The two drugs also reduce erection. Penile erection elicited by papaverine is reduced profoundly in mice with targeted deletion of eNOS. Our findings support a model in which rapid, brief activation of neuronal NOS initiates the erectile process, whereas PI3-kinase/Akt-dependent phosphorylation and activation of eNOS leads to sustained NO production and maximal erection. PMID- 11904452 TI - AtKC1, a silent Arabidopsis potassium channel alpha -subunit modulates root hair K+ influx. AB - Ion channels in roots allow the plant to gain access to nutrients. The composition of the individual ion channels and the functional contribution of different alpha-subunits is largely unknown. Focusing on K(+)-selective ion channels, we have characterized AtKC1, a new alpha-subunit from the Arabidopsis shaker-like ion channel family. Promoter-beta-glucuronidase (GUS) studies identified AtKC1 expression predominantly in root hairs and root endodermis. Specific antibodies recognized AtKC1 at the plasma membrane. To analyze further the abundance and the functional contribution of the different K(+) channels alpha-subunits in root cells, we performed real-time reverse transcription-PCR and patch-clamp experiments on isolated root hair protoplasts. Studying all shaker-like ion channel alpha-subunits, we only found the K(+) inward rectifier AtKC1 and AKT1 and the K(+) outward rectifier GORK to be expressed in this cell type. Akt1 knockout plants essentially lacked inward rectifying K(+) currents. In contrast, inward rectifying K(+) currents were present in AtKC1 knockout plants, but fundamentally altered with respect to gating and cation sensitivity. This indicates that the AtKC1 alpha-subunit represents an integral component of functional root hair K(+) uptake channels. PMID- 11904453 TI - Functional asymmetry of photosystem II D1 and D2 peripheral chlorophyll mutants of Chlamydomonas reinhardtii. AB - The peripheral accessory chlorophylls (Chls) of the photosystem II (PSII) reaction center (RC) are coordinated by a pair of symmetry-related histidine residues (D1-H118 and D2-H117). These Chls participate in energy transfer from the proximal antennae complexes (CP43 and CP47) to the RC core chromophores. In addition, one or both of the peripheral Chls are redox-active and participate in a low-quantum-yield electron transfer cycle around PSII. We demonstrate that conservative mutations of the D2-H117 residue result in decreased Chl fluorescence quenching efficiency attributed to reduced accumulation of the peripheral accessory Chl cation, Chl(Z)(+). In contrast, identical symmetry related mutations at residue D1-H118 had no effect on Chl fluorescence yield or quenching kinetics. Mutagenesis of the D2-H117 residue also altered the line width of the Chl(Z)(+) EPR signal, but the line shape of the D1-H118Q mutant remained unchanged. The D1-H118 and D2-H117 mutations also altered energy transfer properties in PSII RCs. Unlike wild type or the D1-H118Q mutant, D2 H117N RCs exhibited a reduced CD doublet in the red region of Chl absorbance band, indicative of reduced energetic coupling between P680 and the peripheral accessory Chl. In addition, transient absorption measurements of D2-H117N RCs, excited on the blue side of the Chl absorbance band, exhibited a ( approximately 400 fs) pheophytin Q(X) band bleach lifetime component not seen in wild-type or D1-H118Q RCs. The origin of this component may be related to delayed fast-energy equilibration of the excited state between the core pigments of this mutant. PMID- 11904454 TI - Integration of emotion and cognition in the lateral prefrontal cortex. AB - We used functional MRI to test the hypothesis that emotional states can selectively influence cognition-related neural activity in lateral prefrontal cortex (PFC), as evidence for an integration of emotion and cognition. Participants (n = 14) watched short videos intended to induce emotional states (pleasant/approach related, unpleasant/withdrawal related, or neutral). After each video, the participants were scanned while performing a 3-back working memory task having either words or faces as stimuli. Task-related neural activity in bilateral PFC showed a predicted pattern: an Emotion x Stimulus crossover interaction, with no main effects, with activity predicting task performance. This highly specific result indicates that emotion and higher cognition can be truly integrated, i.e., at some point of processing, functional specialization is lost, and emotion and cognition conjointly and equally contribute to the control of thought and behavior. Other regions in lateral PFC showed hemispheric specialization for emotion and for stimuli separately, consistent with a hierarchical and hemisphere-based mechanism of integration. PMID- 11904455 TI - Fear recognition in the voice is modulated by unconsciously recognized facial expressions but not by unconsciously recognized affective pictures. AB - Multisensory integration is a powerful mechanism for increasing adaptive responses, as illustrated by binding of fear expressed in a face with fear present in a voice. To understand the role of awareness in intersensory integration of affective information we studied multisensory integration under conditions of conscious and nonconscious processing of the visual component of an audiovisual stimulus pair. Auditory-event-related potentials were measured in two patients (GY and DB) who were unable to perceive visual stimuli consciously because of striate cortex damage. To explore the role of conscious vision of audiovisual pairing, we also compared audiovisual integration in either naturalistic pairings (a facial expression paired with an emotional voice) or semantic pairings (an emotional picture paired with the same voice). We studied the hypothesis that semantic pairings, unlike naturalistic pairings, might require mediation by intact visual cortex and possibly by feedback to primary cortex from higher cognitive processes. Our results indicate that presenting incongruent visual affective information together with the voice translates as an amplitude decrease of auditory-event-related potentials. This effect obtains for both naturalistic and semantic pairings in the intact field, but is restricted to the naturalistic pairings in the blind field. PMID- 11904457 TI - Argosomes: intracellular transport vehicles for intercellular signals? AB - Cell clusters in the immature tissues of developing organisms create morphogen gradients that guide cellular differentiation into specific cell fates. Although the process of simple diffusion in gradient establishment has been well studied, there are other mechanisms by which cells establish morphogen gradients. Christian discusses the recent findings that morphogens may establish gradients through the use of plasma membrane-containing exovesicles, termed "argosomes." PMID- 11904456 TI - The perceptual reality of synesthetic colors. AB - Synesthesia is a remarkable, rare condition where an individual has multimodal perceptual experiences from a unimodal sensory event. We have studied such an individual, an adult male for whom achromatic words and alphanumeric characters are seen in vivid, reliable colors. We used a variety of perceptual tasks to document the perceptual reality of synesthetic colors and to begin to localize the stage of visual processing where this anomalous binding of externally specified form and internally generated color may take place. Synesthetic colors were elicited by forms defined solely by binocular cues or solely by motion cues, which implies a central locus of visual processing for synesthetic binding of form and color. Also included among our measurements was a difficult visual search task on which non-synesthetic subjects required an effortful search through the visual display. Our subject, in contrast to non-synesthetic subjects, accomplished the task with relative ease because the target of the search had a different synesthetic color from the distractors. Thus, synesthetic experiences appear to originate from a binding of color and form that takes place within central stages of visual processing. PMID- 11904458 TI - Dancing with multiple partners. AB - Transmembrane proteins, such as G protein-coupled receptors (GPCRs) and integrins, activate intracellular signaling pathways through interactions with downstream binding partners. Woodside discusses two examples in which GPCRs and integrins interact in a noncompeting manner with more than one partner. The specific GPCR described is the thrombin receptor, in experiments where G protein peptides selectively block signaling through a particular G protein that does not appear to inhibit coupling of the receptor to other G proteins. The second system described is the alphaIIbbeta3 integrin and its activation of the nonreceptor tyrosine kinase Syk. Syk appeared capable of interacting with both the integrin and intracellular domains of immune response receptors, because binding of Syk to the integrin was not inhibited by peptides based on the Syk binding site in immune response receptors. Thus, multiple, noncompeting binding partners add to the complexity of signal transduction outputs from a single receptor complex. PMID- 11904459 TI - Fibroblast growth factor receptor 3 (FGFR3) - analyses of the S249C mutation and protein expression in primary cervical carcinomas. AB - Fibroblast growth factor receptor 3 (FGFR3) seems to play an inhibitory role in bone development, as activating mutations in the gene underlie disorders such as achondroplasia and thanatophoric dysplasia. Findings from multiple myeloma (MM) indicate that FGFR3 also can act as an oncogene, and mutation of codon 249 in the fibroblast growth factor receptor 3 (FGFR3) gene was recently detected in 3/12 primary cervical carcinomas. We have analysed 91 cervical carcinomas for this specific S249C mutation using amplification created restriction site methodology (ACRS), and detected no mutations. Immunohistochemistry was performed on 73 of the tumours. Reduced protein staining was seen in 43 (58.8%) samples. Six of the tumours (8.2%) revealed increased protein staining compared with normal cervical tissue. These patients had a better prognosis than those with reduced or normal levels, although not statistically significant. This report weakens the hypothesis of FGFR3 as an oncogene of importance in cervical carcinomas. PMID- 11904460 TI - The biological safety of condom material can be determined using an in vitro cell culture system. AB - Latex products have long been recognized as a cause of latex protein allergy. The increased usage of latex gloves, with the consequent increased occurrence of latex allergies appears to have escalated with increasing awareness of the transmission of HIV-AIDS and other infections. The use of condoms as a means to prevent the transmission of STD's (sexually transmitted diseases) and HIV-AIDS has been widely promoted. Although extensive testing is done to evaluate the physical quality of condoms, no information is available regarding the biological safety of condoms. This study was undertaken to determine the effects of short term exposure to physiological levels of condom surface material on cell viability (MTT assay) and cell growth (crystal violet assay). A direct contact cell culture testing method (FDA test method F813-83 used to evaluate the cytotoxic potential of medical materials and devices) was used. The modified test method was found to be a sensitive test system for the evaluation of the biological safety of condoms. This study reveals the importance of evaluating the biological safety of all condoms that are commercially available, because of the potential health risk that may be associated with prolonged use of certain types of condoms. PMID- 11904461 TI - High resolution comparative genomic hybridization detects 7-8 megabasepair deletion in PCR amplified DNA. AB - We investigated if any change in spatial resolution of comparative genomic hybridization analysis could be detected when using DNA amplified by degenerate oligonucleotide primed PCR (DOP-PCR) as opposed to the use of unamplified DNA. Five DNA samples from B-cell leukemias with small 11q deletions were amplified by DOP-PCR and analysed by means of high resolution comparative genomic hybridization (HR-CGH) for the evaluation of aberration size detection limit. By means of HR-CGH, we found the detection limit of DOP-PCR CGH for deletions to be between 3 Mbp and 7-8 Mbp. PMID- 11904462 TI - Topical mitomycin C and radiation induce conjunctival DNA-polyploidy. AB - INTRODUCTION: Atypical cell changes often occur following treatment of premalignant or malignant conjunctival neoplasias with topical mitomycin C (MMC) and/or radiation. These reactive, non-neoplastic alterations of the conjunctival epithelium can be a differential diagnostic problem. Our aim was to investigate changes in the nuclear DNA-distribution of conjunctival epithelial cells after MMC- and radiation therapy by DNA-image-cytometry. METHODS: Conjunctival brush smears were obtained from 13 patients (13 eyes) with squamous cell carcinomas and six patients (6 eyes) with conjunctival malignant melanomas in situ before, during and after treatment. The patients were treated with MMC-drops (0.02% or 0.04%) alone (n=12), with radiation therapy (n=3) or both (n=4). At first, the obtained brush smears were evaluated by cytology. Secondly, after Feulgen restaining, the DNA content of reactively changed cells was determined using the AutoCyte-QUIC-DNA workstation. RESULTS: We observed euploid DNA-polyploidy and cytomorphological changes in all patients (19/19). We considered these alterations as reactive to treatment. Four patients showed their greatest DNA stemline at 4c and 15 patients at 8c. This effect was observed during and following MMC-drops and/or radiation and remained stable in 94% of all patients after a mean follow-up of 22.5 months (SD 15.4). In five cases image cytometry additionally demonstrated DNA-stemline aneuploidy as an evidence of tumor recurrence. CONCLUSION: Measurements of DNA-content revealed euploid polyploidisation of morphological suspicious but benign squamous cells which is the biologic correlate of well known secondary morphologic changes following topical chemotherapy and/or radiation. DNA-image-cytometry is a useful tool in the differention of euploid polyploidization as a sign of reactive cell changes following treatment and tumor recurrences. PMID- 11904463 TI - Prognostic classification of early ovarian cancer based on very low dimensionality adaptive texture feature vectors from cell nuclei from monolayers and histological sections. AB - In order to study the prognostic value of quantifying the chromatin structure of cell nuclei from patients with early ovarian cancer, low dimensionality adaptive fractal and Gray Level Cooccurrence Matrix texture feature vectors were extracted from nuclei images of monolayers and histological sections. Each light microscopy nucleus image was divided into a peripheral and a central part, representing 30% and 70% of the total area of the nucleus, respectively. Textural features were then extracted from the peripheral and central parts of the nuclei images.The adaptive feature extraction was based on Class Difference Matrices and Class Distance Matrices. These matrices were useful to illustrate the difference in chromatin texture between the good and bad prognosis classes of ovarian samples. Class Difference and Distance Matrices also clearly illustrated the difference in texture between the peripheral and central parts of cell nuclei. Both when working with nuclei images from monolayers and from histological sections it seems useful to extract separate features from the peripheral and central parts of the nuclei images. PMID- 11904464 TI - Fourth updated ESACP consensus report on diagnostic DNA image cytometry. AB - A task force of experts in the field of diagnostic DNA image cytometry, invited by the ESACP, and further scientists or physicians revealing experience in that diagnostic procedure (names are given in Addendum A), agreed upon the following 4th updated Consensus Report on Standardised Diagnostic DNA Image Cytometry during the 7th International Congress of that society in Caen, 2001. This report is based on the three preceding ones [6,14,17]. It deals with the following items:- Critical review and update of the definitions given in the 1997 Consensus Update;- Review and detailed description of basic terms, principles and algorithms for diagnostic interpretation;- Recommendations concerning diagnostic or prognostic applications in specific fields of tumour pathology. This update is not aimed to substitute the 1997 consensus, but to make necessary addenda and give more detailed descriptions of those items not unequivocally to interpret by potential users of the methodology. PMID- 11904465 TI - Influence of temperature variation from 5 degrees C to 37 degrees C on aggregation and deformability of erythrocytes. AB - Erythrocyte aggregation and elongation index (EI) (deformability) are measured at temperatures ranging from 5 degrees C to 37 degrees C by aggregometer MA1, and diffractometer Rheodyne SSD and the LORCA, respectively. The test samples are prepared from blood obtained from healthy subjects in test tubes containing EDTA as an anticoagulant and prior to measurement are placed in a water bath for 30 min, maintained at respective temperatures. The aggregation process is given in terms of primary aggregation time and indices of aggregation. These show significant to highly significant changes with the increase of temperature compared to that at 5 degrees C. The EI increases with the increase of shear stress but shows significant decrease with the decrease of temperature. PMID- 11904466 TI - Rheological properties of blood in patients with chronic liver disease. AB - We analyzed rheologic parameters, including erythrocyte rigidity (ER), whole blood and plasma viscosity, erythrocyte and platelet count, hemoglobin, hematocrit, mean corpuscular volume (MCV), fibrinogen, erythrocyte sedimentation rate (ESR), cholesterol, triglyceride, high-density lipoprotein (HDL), low density lipoprotein (LDL), very-low density lipoprotein (VLDL), and gamma globulin levels in 18 patients with chronic liver disease and 20 healthy volunteers. Fifteen patients had cryptogenic cirrhosis while 3 had chronic active hepatitis. ER and MCV was significantly higher in the patient group than the control group while whole blood and plasma viscosities were significantly lower. There were significant correlations between ER and blood and plasma viscosity, ER and MCV, plasma and blood viscosity, HDL and plasma viscosity and a negative correlation between ER and ESR. Our results demonstrate that erythrocytes become more rigid in chronic liver disease. We suggest that erythrocytes with increased rigidity can impair hepatic microvascular circulation and thus contribute to liver dysfunction. PMID- 11904467 TI - Cerebral microcirculatory changes in rat with a cardiopulmonary bypass using fluorescence videomicroscopy. AB - Cerebral microcirculatory changes in rat with a cardiopulmonary bypass (CPB) at normothermia was investigated in relation to cerebrovascular disorders caused by surgical operation with CPB. The mean arterial pressure was changed from 50 to 200 mmHg by changing the pump flow-rate. A non-pulsatile flow model was developed by stopping the cardiac beat using a fibrillator. The pial microcirculation was visualized using fluorescence-labeled red cells and dextran, and was directly observed under a fluorescence videomicroscope during CPB. Based on the recorded videoimages, the arteriolar diameter and red cell velocity were measured, in which single arterioles with approximately 40 microm diameter were selected among the pial arterioles. It was shown that when the arterial pressure was changed: (1) arteriolar vasodilation or constriction appeared during pulsatile flow but it disappeared during non-pulsatile flow, and (2) the arteriolar red cell velocity increased or decreased linearly during non-pulsatile flow as well as pulsatile flow. The flow-rate was almost constant at a large range of the mean arterial pressure from 60 to 160 mmHg during pulsatile flow (autoregulation), but it increased or decreased during non-pulsatile flow with an increase or decrease in mean arterial pressure, respectively. It was suggested that pulsativity might be responsible for cerebral autoregulation. PMID- 11904468 TI - Red blood cell rheological properties in various rat hypertension models. AB - Red blood cell (RBC) properties were proposed to play role in the development of hypertension (HT). This study aimed at investigating the alterations of RBC deformability and aggregation, in various models of HT in rats. The following four models of HT were developed in rats: one kidney-one clip HT (1K-1C HT), two kidney-one clip HT (2K-1C HT), deoxycorticosterone acetate (DOCA) induced HT (15 mg/kg, 2 times/week, sc) and N-nitro L-arginine methyl ester (L-NAME) induced HT (50 mg/kg/day, 10 weeks, ip). The blood samples were obtained from abdominal aorta, under ether anesthesia, after a period of 10 weeks of increased blood pressure. RBC deformability was determined by ektacytometry. RBC aggregation was measured in autologous plasma and 0.5% dextran 500, using a photometric rheoscope. Plasma fibrinogen concentration was determined by Clauss method. The mean blood pressure in all four HT models were about 140 mmHg, on the day of sampling, compared to approximately 110 mmHg in the control group. RBC deformability was found to be significantly decreased in the L-NAME model of HT. RBC aggregation in autologous plasma was significantly higher than control in 2K 1C, L-NAME and DOCA models, DOCA HT model being the most effective in altering the RBC aggregation. Plasma fibrinogen values were found to be higher than control in 2K-1C and L-NAME HT models, but not in DOCA HT. These results confirm that RBC rheological properties might be altered in HT. It can also be suggested that these alternations may not simply be the result of the vascular effects of HT, but may play role in the development of HT, as the alterations in different HT models were not the same, although the length and magnitude of increased blood pressure were similar. PMID- 11904469 TI - Activity of heparins in experimental models of malignancy: Possible explanations for anticancer effects in humans. AB - Heparin and heparin-like compounds appear to possess anticancer properties apart from their anticoagulant activities. This paper reviews recent data on heparins in experimental models of tumor growth and metastasis and discusses various mechanisms by which heparins may inhibit cancer progression. The growing body of evidence supporting the antineoplastic activity of heparins provides the rationale for their widespread testing in cancer patients for the purpose of improving cancer-related survival. Their improved safety, convenience and ease of outpatient administration compared to unfractionated heparin, as well as the suggestion of superior anticancer activity, make the low molecular weight heparins the preferred agents to test in prospective cancer trials. PMID- 11904470 TI - Biological risk factors for deep vein trombosis. AB - Hypercoagulable states due either to inherited or acquired thrombotic risk factors are only present in approximately half of cases of DVT, but the causes in the other half, remain unknown. The importance of biological risk factors such as hyperlipidemia, hypofibrinolysis and hemorheological alterations in the pathogenesis of DVT has not been well established. In order to ascertain whether the above mentioned biological factors are associated with DVT and could constitute independent risk factors, we carried out a case-control study in 109 first DVT patients in whom inherited or acquired thrombophilic risk factors had been ruled out and 121 healthy controls age (42+/-15 years) and sex matched. From all the biological variables analyzed (cholesterol, triglycerides, glucose, fibrinogen, erythrocyte aggregation, hematocrit, plasma viscosity and PAI-1) only fibrinogen concentration reached a statistically significant difference on the comparison of means (290+/-73 mg/dl in cases vs 268+/-58 mg/dl in controls, p<0.05). After this continuous variables were dichotomized according to our reference values, the percentage of cases with cholesterolemia >220 mg/dl, hematocrit >45% and fibrinogen >300 mg/dl was higher in cases than in controls: 38% vs 22%; p<0.01; 43% vs 27%; p<0.05; 36% vs 23%; p<0.05, respectively. The percentage of cases with PAI-1 values >30 ng/ml, 37% vs 25% was borderline significant; p=0.055. Multivariate logistic regression analysis showed that cholesterolemia >220 mg/dl and fibrinogen >300 mg/dl constitute independent predictors of venous thrombotic risk. The adjusted OR's were 2.03 (95% CI; 1.12 3.70) for cholesterolemia and 1.94 (95% CI; 1.07-3.55) for fibrinogen. When these two variables combined DVT risk rose about fourfold (3.96; p<0.05). Our results suggest that hypercholesterolemia and hyperfibrinogenemia should be added to the list of known DVT risk factors and we recommend adopting measures to decrease these variables in the population with a high risk of DVT. PMID- 11904471 TI - Prognostic role of plasmaviscosity in breast cancer. AB - Tumor growth leads to tissue hypoxia and tissue hypoxia, in turn, is a strong stimulus for expression of genes encoding factors that promote tumor growth. Likewise, hypoxia is a key condition within the vicious cycle of autogenous neoplastic dissemination. A marker of the presence of tissue hypoxia may be the presence of high blood viscosity, which is found in a number of neoplastic diseases including gynecological cancer. At the time of diagnosis of breast cancer, patients dying of this disease had had significantly higher initial pv (1.40+/-0.18 mPa s; p<0.0001) when compared to patients not dying of cancer (1.30+/-0.10 mPa s). In multivariate proportional hazard regression analysis, next to tumor size (p=0.03) and nodal status (p=0.004), pv was found an independent prognostic marker for overall survival of breast cancer patients (RR=130.2; 95% CI: 11.6-1,460.6; p<0.0001). An optimized preoperative cut-off value above 1.40 mPa s was significantly associated with poor outcome (log-rank test) in the Kaplan-Meier survival-estimates, even in node-negative breast cancer (n=153; 54.6%). The most likely explanation for these findings is that increased fibrinogen/fibrin turnover and breakdown products characteristically associated with tumor-cell dissemination contribute to the increased plasma viscosity while the hematocrit, leukocyte count, and platelet count contributed little to the increased blood viscosity in patients with breast cancer. These findings may constitute an approach for new strategies in cancer therapy since it might be possible that reduction of plasma viscosity by treatment improves responsiveness to radio/chemotherapy and thus survival of patients. PMID- 11904472 TI - [Psychosocial and psychosomatic basic competence of the gynecologist--from intrinsic conviction to a learnable curriculum]. AB - The gynecologist is confronted with many tasks for which she/he needs psychosocial competence: patient education and health promotion, counselling, and management of psychosocial problems during the various phases of a woman's life cycle, care for patients with chronic diseases. To obtain this competence, a curriculum is needed which comprises elements of psychology, psychosocial medicine, and psychiatry adapted to the specific needs in women's health care. Knowledge and skills derived from communications theory and practice include the semiotic and pragmatic model of communication, physician- and patient-centered communication, counselling for health behavior changes, breaking bad and sad news, coping, and supportive counselling. PMID- 11904473 TI - [Chemotherapy-associated myelosuppression in gynecological oncology]. AB - INTRODUCTION AND OBJECTIVE: A clinically important myelosuppression due to adjuvant chemotherapy is seen more frequently as dosage is intensified and new drugs are used. The assessment of the cytopenia expected is frequently hampered by a lack of directly comparable data. The aim of this study was to compare - in our own patient population - the chemotherapy-associated myelosuppression of four chemotherapeutic regimens used in gynecological oncology. METHODS: 79 patients with primary breast cancer and 26 patients with epithelial ovarian carcinoma underwent cytostatic treatment, and the associated myelosuppression was evaluated by the determination of cytopenia and the need for supportive therapy. The chemotherapy regimens investigated were CMF (cyclophosphamide 600 mg/m(2), methotrexate 40 mg/m(2), 5-fluorouracil 600 mg/m(2), 6xq3w), EC/CMF (epirubicin 90 mg/m(2), cyclophosphamide 600 mg/m(2), 4xq3w, followed by CMF, 3xq3w), DE (docetaxel 75 mg/m(2), epirubicin 90 mg/m(2), 6xq3w) and CC (cyclophosphamide 600 mg/m(2), carboplatin AUC 6, 6xq3w). RESULTS: The EC/CMF and DE regimens were used significantly more frequently for more advanced tumor stages, but there were no differences concerning tumor-dependent prechemotherapeutic myelosuppression. Hemopoiesis was most impaired in the CC group with a mean drop of serum hemoglobin of 1.5 g/dl to the end of the cytostatic treatment; correspondingly, most transfusions of concentrated erythrocytes were needed in this group. The strongest suppression of leukopoiesis was found in the DE group, with a mean drop in leukocyte counts of 6.2 x 10(3)/microliter per cycle; in this group, a mean of 7.6 ready-made syringes with 263 microgram Lenogastrim was used to stimulate leukopoiesis. The severest drop in the mean thrombocyte count, i.e. 171.7 x 10(3)/microliter, was found in the CC group. CONCLUSIONS: The CC regimen impairs thrombo- and erythropoiesis most, whereas the DE regimen causes marked leukopenia. The regimen with the smallest myelosuppression was CMF. No severe cytopenia-associated complications were detected in any of the cases investigated. PMID- 11904474 TI - [Bumm, Zweifel, Kubli and others--examples of the academic interlacing between Switzerland and Germany]. AB - There is an interlacing in academic gynecology and obstetrics between Germany and Switzerland which leads back to the founding years of both societies. Germans have been elected to the chairs in Zurich, Bern, and Basel, whereas Swiss have been appointed to Erlangen, Leipzig, Dresden, and Frankfurt. Reservations from both sides, if any, were not prevailing. Even if the balance at present might show a trend again towards the Germans being chairmen at German-speaking Swiss Universities, the historical line contradicts the impression of one-sidedness. It is the academic exchange followed by a remarkable interlacing between the two German-speaking nations which will certainly continue into the future, all the more so in a Europe where historical differences are more or less overcome. The younger generation will extend and improve the links already laid down in the past. The men mentioned above have all been leading figures during their time, Fred Kubli so in particular. They will be followed by successors. Might they be as excellent as their predecessors have been. PMID- 11904475 TI - [Guideline for clarification of the palpable breast finding]. PMID- 11904476 TI - [Guidelines for diagnosis and therapy of intraepithelial neoplasia and early invasive carcinoma of the female lower genital system (cervix uteri, vagina, vulva) established by the AGK (Colposcopy Work Group in the OGGG [Austrian Society of Gynecology and Obstetrics])]. PMID- 11904520 TI - Evidence that lacidipine at nonsustained antihypertensive doses activates nitrogen monoxide system in the endothelium of salt-loaded Dahl-S rats. AB - Lacidipine is a clinically active, antihypertensive calcium antagonist of the 1,4 dihydropyridine (DHP) class. It is also capable of vascular protection when administered (prophylactically and therapeutically) at nonsustained antihypertensive doses to salt-sensitive Dahl-S rats: useful animal models for studying the vasoprotective effect of calcium antagonists. In our previous work using voltammetry with selective biosensors, we observed that lacidipine implements endothelial nitrogen monoxide (NO) in normal rats. These experiments, performed in aortic rings obtained from Dahl-S rats analyzed with voltammetry and specific biosensors, further demonstrate that lacidipine, given at doses that do not control the development of hypertension (1 mg/kg), enhance endothelial NO activity. Taken together with the observation that 1 mg/kg lacidipine, and not its vehicle, is able to prevent vascular damage and concomitant increases in mortality (accelerated by a salt diet), this voltametric data suggest that NO acts as a component of positive influence of this DHP on vascular structure and function. PMID- 11904521 TI - Multiple interactions between the renin-angiotensin and the kallikrein-kinin systems: role of ACE inhibition and AT1 receptor blockade. AB - The investigation of therapeutic actions of angiotensin type 1 (AT1) receptor antagonists and ACE inhibitors (ACEI) demonstrated complex interactions between the renin-angiotensin system (RAS) and the kallikrein-kinin system (KKS) in several experimental and clinical studies. They are evidenced by the fact that (1) ACE efficiently catabolizes kinins; (2) angiotensin-derivatives such as ANG (1-7) exert kininlike effects; and (3) kallikrein probably serves as a prorenin activating enzyme. (4) Several authors have demonstrated experimentally that the protective effects of ACEI are at least partly mediated by a direct potentiation of kinin receptor response on BK stimulation. (5) Furthermore, studies on AT1 antagonists, which do not directly influence kinin degradation, and studies on angiotensin-receptor transgenic mice have revealed additional interactions between the RAS and the KKS. There is mounting evidence that an autocrine cascade including kinins, nitric oxide, prostaglandins, and cyclic GMP is involved in at least some of the angiotensin type 2 receptor effects. This review discusses multiple possibilities of cross-talks between the RAS and KKS in vascular and cardiac physiology and pathology after ACE inhibition and AT1 receptor blockade. PMID- 11904522 TI - Antihypertensive treatment with enrasentan (SB217242) in an animal model of hypertension and hyperinsulinemia. AB - Enrasentan is an antagonist of endothelin (ET) receptors. Previous studies have shown that antagonism of ET receptors might represent a new approach to the treatment of hypertension. Rats with a high-fructose diet (HFD) develop hyperinsulinemia, hypertriglyceridemia, and hypertension; renal and cardiac damage. The aim of this study was to evaluate whether enrasentan could reverse the hypertension and reduce the target organ damage induced by an HFD. Fifty-five WKY rats were divided into 6 groups; 35 animals received HFD for a month; thereafter 5 animals were killed, and the others were treated either with enrasentan (n = 10), hydralazine (n = 10), or placebo (n = 10) for a further month while on the HFD. Twenty animals were kept on a standard diet throughout the study; either placebo (n = 10) or enrasentan (n = 10) was administered during the second month. Enrasentan and hydralazine completely eliminated the HFD induced increase in blood pressure; however, only enrasentan reduced the renal and cardiac damage caused by the diet. In conclusion, enrasentan was effective both in normalizing blood pressure and in reducing renal and cardiac damage; the organ protection cannot be attributed solely to the antihypertensive effect, because it was absent in the case of hydralazine, despite successful control of blood pressure. PMID- 11904523 TI - Effects of ethinyl estradiol, estradiol, and testosterone on hindlimb endothelial function in vivo. AB - SUMMARY: This study was designed to evaluate the effects of chronic treatment with estrogen and testosterone on hindlimb vascular function. Spontaneously hypertensive rats (SHR) were sham-operated (SHAM) or ovariectomized and treated with vehicle (OVX), ethinyl estradiol (OVX-EE2), estradiol (OVX-E2), or testosterone (OVX-TESTO) for 3 weeks. Anesthetized SHR were instrumented for the measurement of arterial blood pressure and hindlimb blood flow. Ovariectomy had no significant effect on hindlimb resistance. Resting blood pressure (mm Hg) and hindlimb resistance (mm Hg/ml/min/kg) were higher in OVX-TESTO (158 +/- 4 and 4.7 +/- 0.3) than SHAM control (130 +/- 6 and 3.3 +/- 0.4), OVX (139 +/- 6 and 3.4 +/ 0.3), OVX-EE2 (130 +/- 4 and 3.0 +/- 0.3), and OVX-E2 (120 +/- 4 and 3.3 +/- 0.4). The hemodynamic responses to the endothelium-dependent vasodilator acetylcholine (ACh) were similar in SHAM, OVX, OVX-EE2, and OVX-E2. The hindlimb vasodilatory effects of ACh were consistently greater in OVX-TESTO compared with OVX, OVX-EE2, OVX-E2, and SHAM control rats. The hemodynamic responses to the nitric oxide donor, sodium nitroprusside, were similar in all groups. Our results provide evidence of modulatory influence of testosterone on vasomotor function. It is suggested that the enhanced hindlimb endothelial function represents a compensatory mechanism for testosterone-induced hypertension. PMID- 11904524 TI - Retinoids induce the PAI-1 gene expression through tyrosine kinase-dependent pathways in vascular smooth muscle cells. AB - Retinoids exert their pleiotropic effects on several pathophysiologic processes, including neointima formation after experimental vascular injury. Plasminogen activator inhibitor-1 (PAI-1) has been proposed to play an inhibitory role in arterial neointima formation after injury. We examined whether retinoids regulate PAI-1 expression in cultured vascular smooth muscle cells (SMCs). Northern blot analysis showed that all-trans retinoic acid (atRA) and 9-cis retinoic acid (9cRA) increased PAI-1 mRNA levels in a dose-dependent manner. These responses were completely inhibited by tyrosine kinase inhibitors. The half-life of PAI-1 was not affected by atRA, suggesting that induction of PAI-1 mRNA was mainly regulated at the transcriptional levels. Stable and transient transfection assays of the human PAI-1 promoter-luciferase constructs indicate that DNA sequence responsive to either ligand-stimulated or overexpressed retinoic acid receptor alpha expression vector lies downstream of -363 relative to the transcription start site, where no putative retinoic acid response element is found. These results indicate that atRA and 9cRA increase PAI-1 gene transcription through pathways involving tyrosine kinases in SMCs. Because PAI-1 inhibits the production of fibrinolytic protein plasmin that facilitates SMC migration, induction of the PAI-1 gene expression by atRA may at least partly account for the role of atRA as an important inhibitor of neointima formation. PMID- 11904526 TI - Effects of combined therapy with clopidogrel and acetylsalicylic acid on platelet glycoprotein expression and aggregation. AB - SUMMARY: This study aimed to compare the effects of clopidogrel, acetylsalicylic acid (ASA), and the combination of both substances on platelet aggregation and expression of platelet membrane glycoproteins in patients with chronic coronary artery disease. We investigated platelet activation by flow cytometry and by platelet aggregation and disaggregation in 60 patients randomly assigned to 3 treatment groups: ASA, clopidogrel, combination of clopidogrel and ASA, treated for 14 days. Adenosine diphosphate (ADP)-induced expression of P-selectin and of PAC-1 was significantly reduced after 2 wk of clopidogrel but not of ASA treatment. Treatment with clopidogrel reduced the ADP-induced platelet aggregation. The combination of clopidogrel and ASA did not increase the inhibition of platelet activation compared with clopidogrel alone. A significant increase in platelet disaggregation was observed with clopidogrel alone and was more pronounced with the combination of clopidogrel and ASA. ADP-induced platelet degranulation, activation of GPIIb/IIIa receptor, and aggregation in vivo are effectively inhibited by clopidogrel. The significantly increased disaggregation under clopidogrel and ASA suggests that the combined therapy may be superior to the monotherapy in patients with coronary artery disease and a high risk for vascular events. PMID- 11904527 TI - Use of A-192621 and IRL-2500 to unmask the mesenteric and renal vasodilator role of endothelin ET(B) receptors. AB - Endothelin-1 (ET-1) is known to cause a transient (<1 min) depressor followed by a sustained (>1 h) pressor response. The former through the activation of ET(B) receptors, and the latter through the activation of ET(A) and ET(B) receptors. This study examines if ET(B) receptors mediate sustained mesenteric and renal dilation in anesthetized rats. Intravenous bolus ET-1 (0.8, 1.4, and 2 nmol/kg) and IRL-1620 (ET(B) agonist, 2, 5, and 10 nmol/kg) caused transient decrease followed by sustained increases in mean arterial pressure (MAP) that were accompanied by increases in total peripheral resistance (TPR), reductions in cardiac output (CO), and mesenteric and renal vasoconstriction. Pretreatment with FR-139317 (ET(A) antagonist, 1 mg/kg) attenuated the pressor and constrictor effects of ET-1 but did not alter responses to IRL-1620. IRL-2500 (ET(B) antagonist, 5 mg/kg) slightly inhibited the renal constrictor effect of IRL-1620, whereas A-192621 (ET(B) antagonist, 5 mg/kg) abolished all hemodynamic responses to IRL-1620. Both IRL-2500 and A-192621 markedly enhanced MAP, TPR, and mesenteric, and the renal constrictor effects of ET-1. Therefore, A-192621 was more effective than IRL-2500 in blocking IRL-1620-induced vasoconstriction, but both augmented constrictor responses to ET-1. The potentiation of ET-1-induced vasoconstriction by ET(B) receptor antagonists revealed a sustained vasodilator role of ET(B) receptors. PMID- 11904528 TI - Dexamethasone inhibits the inducible bioconversion of glyceryl trinitrate to nitric oxide. AB - The aim of this study was to assess the effects of dexamethasone (DEX) on the inducible bioconversion of glyceryl trinitrate (GTN) into nitric oxide in cultured smooth muscle cells, endothelial cells, and the J774 macrophage cell line as well as in vivo and ex vivo in rats either untreated or pretreated with Escherichia coli lipopolysaccharide. In vitro, an increased bioconversion of GTN to nitrite and an elevation of cyclosine guanosine 3,5;-monophosphate (cGMP) levels occurred after treatment with lipopolysaccharide (LPS) (0.5 microg/ml, 18 h). This effect was ablated by co-incubation with DEX (10 microM, 18 h). Rats treated with an intraperitoneal (IP) injection of LPS (4 mg/kg) 18 h beforehand showed enhanced hypotensive responses to GTN (1 mg/kg, intravenously [IV]) and this was prevented when DEX (4 mg/kg, IP) was given together with LPS. Progesterone (50 mg/kg, IP) had no effect on GTN-induced hypotensive response. Conversely, exposure of rat aortic strips obtained from animals pretreated with LPS produced an enhanced vasorelaxant response in LPS-treated rats. Also, this effect was inhibited by pretreatment with DEX. Thus, the induction of the pathway leading to the formation of nitric oxide from GTN is blocked by DEX both in vitro and in vivo, and this may represent a useful tool in the assessment of the enhanced bioconversion of organic nitrates into nitric oxide occurring via inflammatory mechanisms. PMID- 11904529 TI - Comparative efficacy between the glycoprotein IIb/IIIa antagonists roxifiban and orbofiban in inhibiting platelet responses in flow models of thrombosis. AB - This study was undertaken to compare the platelet binding characteristics and anti-platelet efficacy of a nonpeptide glycoprotein IIb/IIIa antagonist roxifiban with orbofiban in static and dynamic adhesion and aggregation assays. The results indicate that roxifiban binds with higher affinity to glycoprotein IIb/IIIa receptors and exhibits slower dissociation rates than orbofiban. Furthermore, the platelet inhibitory effects of roxifiban, but not orbofiban, were unaffected by changes in plasma calcium concentrations. Both agents reduced, in a concentration dependent manner, the size of platelet thrombi deposited onto collagen I upon perfusion of heparinized blood at a shear rate of 1,500/s. At a clinically achievable concentration of 60 nM, roxifiban abrogated the formation of thrombi containing > 20 platelets per thrombus, thereby displaying comparable in vitro efficacy to that achieved by the theoretical maximal abciximab blood concentration (3.5 microg/ml) produced after standard treatment. In contrast, orbofiban, even at 500 nM, was only effective in inhibiting the formation of larger platelet thrombi (> or =150 platelets per thrombus). Pretreatment of surface-anchored platelets with roxifiban (100 nM), but not orbofiban (500 nM), inhibited monocytic THP-1 cell attachment under flow. However, this heterotypic adhesion process was also suppressed when orbofiban (500 nM) was maintained in the perfusion buffer during the entire course of flow experiment. These findings demonstrate roxifiban (unlike orbofiban) is a potent glycoprotein IIb/IIIa antagonist with a long receptor-bound lifetime and prolonged anti-platelet efficacy and may thus be beneficial for the treatment and prevention of acute ischemic syndromes. PMID- 11904525 TI - Reduction in neointimal formation with a stent coated with multiple layers of releasable heparin in porcine coronary arteries. AB - Recent studies demonstrated that neointimal formation, which is caused by both neointimal proliferation and organized mural thrombus, is responsible for in stent restenosis. Although various types of heparin coatings were effective in reducing (sub)acute thrombosis, most of them failed to reduce neointimal proliferation. This study was designed to examine the effect of the stent coated with multiple layers of releasable heparin complex from which heparin diffuses into the surrounding tissue and exerts its beneficial effects. Male Yorkshire pigs underwent balloon expandable stenting for coronary segments of both the left anterior and the left circumflex coronary arteries with a comparable diameter (n = 10). The stent implantation site was randomized for either control or heparin coated stent. Four weeks after the procedure, quantitative coronary angiography (QCA) and intravascular ultrasonographic imaging (IVUS) were performed followed by histologic analysis. In additional animals, staining for proliferating cell nuclear antigen (PCNA) was performed 10 d after the procedure (n = 3). QCA demonstrated that coronary diameter (mm) was significantly larger at the heparin coated stent site (2.32 +/- 0.14) compared with the control stent site (1.81 +/- 0.17) (p < 0.01). IVUS also showed that the neointimal area (mm2) was significantly suppressed at the heparin-coated stent site (2.12 +/- 0.58) compared with the control stent site (3.92 +/- 0.33) (p < 0.01). Histologic analysis also demonstrated that neointimal area (mm2) was significantly less at the heparin-coated stent (2.94 +/- 0.43) than at the control stent site (4.41 +/- 0.38) (p < 0.01), which was also the case for organized thrombus area (x10-4 mm2) (6.61 +/- 2.67 vs. 19.36 +/- 4.38, p < 0.01). The frequency of PCNA-positive vascular smooth muscle cells (%) was significantly less at the heparin-coated stent (10.8 +/- 1.0) than at the control stent site (19.1 +/- 1.7) (p < 0.01). These results suggest that the stent coated with releasable heparin is beneficial in reducing neointimal formation and subsequent in-stent restenosis. PMID- 11904531 TI - Protective effect of Na+ /H+ exchange inhibitor, SM-20550, on impaired mitochondrial respiratory function and mitochondrial Ca2+ overload in ischemic/reperfused rat hearts. AB - The aim of this study was to investigate whether a selective Na+/H+ exchange inhibitor, SM-20550, can modulate the mitochondrial respiratory function and mitochondrial Ca2+ content in isolated rat hearts subjected to 40 min of ischemia and 20 min of reperfusion. SM-20550 (10, 100 nM) was administered for 5 min prior to ischemia and for 20 min during the reperfusion period. At 20 min after reperfusion, treatment with SM-20550 (10, 100 nM) improved the recovery of left ventricular developed pressure and suppressed the rise in left ventricular end diastolic pressure. Mitochondrial function, assessed by the state 3 oxygen respiration rate, respiratory control index, and oxidative phosphorylation rate, was significantly impaired after ischemia/reperfusion. Administration with SM 20550 (10, 100 nM) attenuated the impaired mitochondrial function, improving the state 3 respiration rate, respiratory control index, and oxidative phosphorylation rate. The mitochondrial Ca2+ content was significantly increased after ischemia/reperfusion but was suppressed by treatment with SM-20550 (10, 100 nM). A significant linear correlation was observed between the respiratory control index and mitochondrial Ca2+ content in the ischemic/reperfused hearts. In conclusion, SM-20550 improved the postischemic recovery of left ventricular function and concurrently protected mitochondrial function mediated by preventing mitochondrial Ca2+ overload. PMID- 11904530 TI - Pharmacodynamic studies on the angiotensin II type 1 antagonists irbesartan and candesartan based on angiotensin II dose response in humans. AB - The in vivo effects of two unsurmountable angiotensin II type 1 (AT1) antagonists, irbesartan (150 mg) and candesartan (8 mg), were studied in a double blind, randomized, crossover study in 18 healthy men. The drugs' direct vascular effects were assessed as the rightward shift (dose ratio - 1) of angiotensin dose effect curves on diastolic blood pressure (DBP). Renal and adrenal effects were assessed by plasma renin activity (PRA), aldosterone concentrations, and antagonistic concentration equivalents (n x Ki) in a radioligand rat lung receptor assay. Both drugs exerted similar substantial (> 30-fold) and long lasting (> 2-fold 47 h after dosing) rightward shifts of the angiotensin II dose effect declining with half-lives of 15 h irbesartan and 12 h candesartan, respectively. Dose ratio - 1 versus n x Ki showed a linear relationship in Schild regression plots; both drugs increased PRA, decreased DBP, and suppressed aldosterone. The slopes of linear relationship between angiotensin antagonism (dose ratio - 1) and PRA increase were almost threefold steeper (p = 0.005) following irbesartan as compared with candesartan. The findings suggest that for the same degree of angiotensin II antagonism, irbesartan produces a greater increase in PRA than candesartan. These pharmacodynamic differences warrant further investigation and clarification. PMID- 11904532 TI - Carvedilol inhibits platelet-derived growth factor-induced signal transduction in human cardiac fibroblasts. AB - In the current study, first the platelet-derived growth factor (PDGF)-induced stimulation of the PDGF-beta receptor kinase in human cardiac fibroblasts was examined, and then the possibility of counterbalancing this signal transduction by carvedilol, a beta-blocker with alpha1-blocking properties, was investigated. Human cardiac fibroblasts were cultured from myocardial biopsy samples taken from patients with idiopathic dilated cardiomyopathy. The stimulation of the PDGF-beta receptor kinase by recombinant human PDGF (BB) in the cells and the inhibitory effect of carvedilol (1, 5, 10, and 20 microM) were investigated by analyzing PDGF-induced PDGF receptor tyrosine phosphorylation using Western blotting and by measuring DNA synthesis with a colorimetric assay. In human cardiac fibroblasts, the PDGF receptor kinase could be stimulated with PDGF (100 ng/ml) and inhibited with carvedilol (5 microM). In addition, carvedilol at a concentration of 5 microM significantly decreased DNA synthesis by approximately 50%. The inhibition of PDGF-stimulated mitogenesis by carvedilol at concentrations of 10 and 20 microM was 64 or 75%, respectively. Other beta-adrenoceptor antagonists such as propranolol (10 microM) and metoprolol (10 microM) did not significantly affect the PDGF-induced beta-receptor autophosphorylation. These findings provide novel experimental support for the known beneficial clinical effects of carvedilol in the treatment of chronic heart failure associated with myocardial fibrosis. PMID- 11904533 TI - Effects of different preproendothelin-1 mRNA anti-sense oligodeoxynucleotides on ischemic arrhythmias in rats. AB - The effects of four anti-sense oligodeoxynucleotides (AS-ODNs) against rat or human preproendothelin-1 mRNA on ischemic arrhythmias in anesthetized rats were studied. AS-ODN (60 nmol/kg) or control (normal saline; sense-ODN, and scrambled ODN, 60 nmol/kg) was injected 2 h before acute myocardial ischemia elicited by the occlusion of the left anterior descending coronary artery. Arrhythmias during 60-min ischemia were assessed, and plasma endothelin-1 was determined with an endothelin-1-specific radioimmunoassay system. The results showed that anti senses against human preproendothelin-1 mRNA were anti-arrhythmic without significant impact on hemodynamics, whereas two against rat preproendothelin-1 mRNA and the three controls failed to be anti-arrhythmic. In human antisense groups, both the incidence of reversible ventricular fibrillation and the mortality were decreased to zero. The incidences of ventricular tachycardia and salvos were significantly decreased from almost 100% in the controls to < or =30% (p < 0.01), the arrhythmia score from an average of approximately 3.6 to 0 and 0.7, respectively (p < 0.01 versus controls), and the total ventricular ectopic beats from an average of 307-338 to < 40 (p < 0.01). The human AS-ODNs led to less plasma endothelin-1, which was associated with suppressed ischemic arrhythmias in this rat model, indicating a contributory role of endothelin-1 in ischemic arrhythmias. Conversely, considering the two- or three-base mismatches between the human AS-ODNs and rat preproendothelin-1 mRNA, and the failure of the rat AS-ODNs in suppressing arrhythmias, the possibility could not be excluded that human endothelin-1 AS-ODNs acted via an undetermined pathway other than endothelin-1. PMID- 11904534 TI - Tissue Angiotensin-converting enzyme activity plays an important role in pressure overload-induced cardiac fibrosis in rats. AB - It has been widely assumed that the cardiac angiotensin-generating system plays an important role in the development and maintenance of cardiac remodeling caused by pressure overload. The roles of angiotensin-converting enzyme (ACE) in pressure overload-induced cardiac hypertrophy and fibrosis in rats were investigated. Pressure overload was achieved by constricting the abdominal aorta above the renal arteries. After they underwent surgery, the rats were treated with a low or high dose of the ACE inhibitor imidapril (0.07 and 0.7 mg/kg/d s.c.) with an osmotic pump for 4 weeks. High-dose imidapril prevented the increase in blood pressure, cardiac hypertrophy, and fibrosis. Low-dose imidapril inhibited only cardiac fibrosis. ACE activity in the myocardium, but not in serum, was significantly increased in the rats with the banded aorta, and ACE immunoreactivity was increased in the areas of fibrosis. These changes were markedly reduced by both doses of imidapril. These results suggest that the increased local ACE expression contributes to the development of pressure overload-induced cardiac fibrosis but is not responsible for hypertrophy in rats. PMID- 11904535 TI - Efficacy of angiotensin II type 1 receptor blockade on reperfusion-induced arrhythmias and mortality early after myocardial infarction is increased in transgenic rats with cardiac angiotensin II type 1 overexpression. AB - Angiotensin II induces ischemia/reperfusion (I/R)-induced arrhythmias and blockade of the angiotensin II type 1 receptor (AT1R) may therefore be beneficial in preventing arrhythmias and decreasing mortality after myocardial infarction (MI). Because the AT1R is upregulated after myocardial ischemia, it was hypothesized that the level of AT1R expression would mediate the response to AT1R blockade. Transgenic (TGR) rats that overexpress the human AT1R and Sprague Dawley rats were used as controls. Total duration of arrhythmia (seconds) after I/R injury was similar in TGR and SD rats (433 +/- 109 vs. 376 +/- 117, p = n.s.). AT1R blockade with losartan decreased total duration of arrhythmia in the TGR rats (433 +/- 110 s-164 +/- 48 s; p < 0.05), whereas it caused a nonsignificant increase in the SD rats (376 +/- 117 s-497 +/- 97). In vivo, survival in the first 24 hours after MI was impaired in TGR rats (39%; SD, 63%). Losartan improved survival significantly in TGR rats (from 39% to 80%, p < 0.05). A smaller, nonsignificant effect was observed in SD rats (63% to 81%). AT1R blockade is beneficial only when the AT1R was overexpressed, both in reducing the reperfusion-induced arrhythmias and mortality early after MI. PMID- 11904546 TI - Drink and drugs: overlooked, underfunded. PMID- 11904547 TI - Faecal impaction: older people's experiences and nursing practice. AB - A number of authors have identified the paucity of evidence available to inform community nursing practice in relation to faecal impaction. Little is known of its incidence or its impact. In an attempt to address this, this article reports part of an extensive descriptive survey in Australia that sought to explore the experience of, and response to, constipation of older people living in the community. The focus here is the participants' experience of extreme constipation in the form of faecal impaction and obstipation (obstinate constipation). Faecal impaction was found to cause great discomfort and distress, often leading to obstipation. The imperative for self-management of constipation was strong, and laxative use frequent. However, manual evacuation was often necessary, either by self or others. This article discusses the results in relation to a recent emergence of concern in the US and UK about how faecal impaction and obstipation should be treated by community nurses. PMID- 11904548 TI - The role of the health visitor in the uptake of cervical smear testing. AB - This article discusses one of the dilemmas health visitors have in promoting the uptake of cervical smear testing. Should health visitors carry out the procedure in the home? In some areas of the country, following many campaigns and efforts the uptake of smear tests remains low whereas in other areas where women are well informed and educated, the uptake is much better. The aim of this article is to give an overview on what has been tried before as well as to discuss the contentious issue of what role the health visitors have in this area of practice. PMID- 11904549 TI - Bandages and bandaging techniques for compression therapy. AB - Bandaging skills are essential for community nurses not only as a method of supporting joints, or retaining a dressing but also as an important treatment of leg ulceration. It is important to be able to choose the correct type, size, and composition of bandage and then apply it safely using the most appropriate technique, as incorrectly applied bandages may lead to pressure necrosis and subsequent limb amputation. Bandaging applications have changed little over the last 100 years and the two most commonly used techniques are still the spiral and the figure-of-eight methods. There are advantages and disadvantages of both, and successful bandaging depends on choosing the correct product and good technique, both in stretching the bandage to the correct tension, and ensuring proper overlap between layers. PMID- 11904550 TI - The extended nurse prescribing curriculum. AB - The extension of independent nurse prescribing is underway; 38 higher education institutions are currently approved to offer the new course. This article discusses the considerations which led to the development of the new curriculum, and provides a broad overview of its content. PMID- 11904551 TI - Glucosamine therapy compared to ibuprofen for joint pain. AB - To determine the effectiveness of oral glucosamine with ibuprofen for the relief of joint pain in osteoarthritis a mini-review (Griffiths, 2002) of double-blind randomized controlled trials comparing the two was undertaken. The population was adult patients diagnosed with osteoarthritis at any site. The outcome was arthritic pain reduction. Searches on Medline, Embase, AMED, the Cochrane Library and the Merck index identified four trials. Of these, two studies were obtainable and were included in the review. Both compared 1.2 g ibuprofen daily with 1.5 g glucosamine sulphate daily, in three divided doses. The combined number of participants in the studies was 218. The results of these studies showed glucosamine to be of similar efficacy to ibuprofen. The conclusion is that glucosamine is effective in relieving joint pain associated with osteoarthritis. Glucosamine's pain-relieving effects may be due to its cartilage-rebuilding properties; these disease-modifying effects are not seen with simple analgesics and are of particular benefit. In practice glucosamine can be used as an alternative to anti-inflammatory drugs and analgesics or as a useful adjunct to standard analgesic therapy. PMID- 11904552 TI - Community care policy and end-of-life care: one patient's story. AB - This article examines the effect of community care policy on the care of a young woman with a terminal illness from a minority ethnic group. Current NHS policy advocates collaboration between health and social care agencies to provide a seamless service for patients. The article considers aspects of interagency and interprofessional collaboration, teamwork and patient empowerment and analyses how beneficial these factors were in a patient's care. It also examines the concept of health promotion within the sphere of palliative care. PMID- 11904553 TI - Stroke patients' carers' views of formal community support. AB - Stroke is a common, disabling condition. Those who look after stroke patients in the community are placed under a lot of strain. In this study a purposive sample of eight carers, chosen to obtain as wide a spread of opinion as possible, were interviewed in depth on their views about the formal support services they receive in the community. A detailed content analysis of the transcripts of the interviews revealed four areas important to carers: information and education; provision of practical help; convenience, coordination and adaptability of service; and consultation with and consideration of the carer. In all areas there was considerable room for improvement. Carers particularly valued community care professionals who listened to them, seemed to understand them and were prepared to put in some effort for them. The importance of ongoing information provision was stressed. PMID- 11904554 TI - Choice and confusion: nurses need to lead. PMID- 11904555 TI - BMA proposes radical changes to the nursing role. PMID- 11904556 TI - Health facilitators in learning disability are important roles. PMID- 11904557 TI - Sexual harassment of female nursing students. PMID- 11904558 TI - Lymphoedema 1: components and function of the lymphatic system. AB - Lymphoedema is an incurable and debilitating condition which has a negative impact on the quality of life of the sufferer and his/her family. Information with regards to diagnosis and treatment is often scarce and conflicting in nature. The following series of articles should enable nurses to recognize the condition, provide basic information to a patient and instigate treatment though referral. The first article describes the anatomy, physiology and functions of the lymphatic system. The focus is on the parts of the lymphatic system which are specific to the condition of lymphoedema and aims to place subsequent articles in context. The following articles describe the different types of lymphoedema and the four main elements of treatment that are central to the management of the condition. PMID- 11904560 TI - Health visitor risk assessment for preventing falls in elderly people. AB - This study was undertaken to look at the feasibility of a health visitor risk assessment for falls at the time of the routine health check for people aged 75 years and above. A total of 162 people were eligible for inclusion in the study. The standard over-75 assessment check was carried out either in the GP surgery or the person's home. A questionnaire was developed to obtain additional information not collected in the routine health check. The results identified two key risk areas: a history of polypharmacy and living in sheltered housing. There were no differences for a range of physical, emotional and environmental factors between people who had fallen and those who had not. A larger study is required to look at the identification of risk factors for falling at the routine over-75 health check, and appropriate referrals that can be put into place to deal with any problems uncovered. Education of health professionals on the risk factors of falls is also required. PMID- 11904559 TI - Collecting pressure ulcer prevention and management outcomes: 2. AB - The first part of this article (Vol 11(4): 230-8) outlined the argument that a combination of efficacy and effectiveness is required to assess fully the impact of interventions such as pressure-redistributing (PR) beds and mattresses. In addition, it described the methodology of this multinational, multicentre, prospective, non-randomized cohort study designed to record the occurrence and characteristics of patients vulnerable to, or with, established pressure ulcers. This article reports further details of the characteristics of the 2507 UK adult hospital patients recruited to the study. Over 40% (42% n = 1046) of all subjects were considered to be at an elevated risk of developing ulcers (Waterlow score of 15 or greater) (Waterlow, 1985). Many were inactive with 332 (13%) confined to bed alone with a further 262 (10%) confined to bed and their chair. Most (74% n = 1868) were nursed upon PR beds and mattresses, while fewer subjects were provided with a PR seat cushion (n = 547; 27%). Two hundred and fifty-seven subjects (10%) experienced at least one change of bed mattress during their stay in hospital, with two subjects being nursed on five different mattresses during their hospital stay. PMID- 11904561 TI - Multidisciplinary care of skin problems in stoma patients. AB - Skin integrity is essential for the normal usage of a stoma appliance. There is little published on the prevalence, prevention or management of stoma skin problems. Allergic contact dermatitis is often cited as the cause, usually without evidence from formal investigations. The authors approached, by postal questionnaire, 525 patients who had had a stoma formation in the last 10 years. A total of 325 responded. All those who described a skin problem were invited to attend a multidisciplinary clinic for further investigations and appropriate treatment of their peristomal skin. This may be severe and debilitating as well as socially restricting. However, with a multidisciplinary approach a number of conditions can be recognized and easily treated, thus improving the quality of life for stoma patients. PMID- 11904562 TI - Legal aspects of consent 23: Department of Health guidelines. AB - This is the last article in the series on the legal aspects of consent. It looks at recent guidance that has been published by the Department of Health on consent for examinations and treatment. It discusses the content of this document and covers the main principles which apply in relation to consent as highlighted in this series. PMID- 11904563 TI - Developing the use of poetry within healthcare culture. AB - While the visual arts are often given consideration as an important feature of healthcare environments, the literary arts remain an underdeveloped resource. This article describes recent nurse-initiated developments in Aberdeen which attempt to integrate poetry into the culture of hospitals so that patients, visitors and staff can be involved. In particular, the 'Poem Post' project is described. This project makes a selection of short poems available on postcards in wall-mounted racks within local hospitals, and incorporates a facility for feedback of comments and new poems. Feedback has been generally very positive and over 100 new poems have been submitted to the project. Issues arising from evaluation of the project are discussed, and lessons learnt from the experience are reviewed in order to encourage nurses to consider the possibilities that poetry can offer in the workplace. PMID- 11904564 TI - Transair Paediatric Mattress replacement system evaluation. AB - While a plethora of pressure-relieving and reducing equipment is available to nurses for the prevention and treatment of pressure ulcers, very little of it is specifically designed or appropriate for paediatric patients. The limited choice that does exist concentrates on neonates or babies, which is problematic for the nurse dealing with the older child. Children may be at increased risk of developing pressure injury as a result of the forces of friction, shearing and pressure that results from being nursed on a support system designed for the adult patient. This gap in service provision may cause dilemmas for the nurse who has a duty to 'cause no harm' (UKCC, 1992) in his/her attempt to protect and treat a child. This article introduces a new product available from Karomed (a division of the Verna Group) that has been specifically designed with the child in mind. PMID- 11904565 TI - The important aspects of nurse specialist roles. PMID- 11904566 TI - People with dementia are not told their diagnosis. PMID- 11904567 TI - The need to reform NHS complaints procedures. PMID- 11904568 TI - Case 56: intimidating behavior. Theatre nurse who behaved inappropriately to staff. PMID- 11904570 TI - Legal aspects of consent 13: organ donation from live donors. AB - Case Scenario: Alice is now 12 years old and has been receiving dialysis for over 3 years. Her long-term chance of survival depends upon a kidney transplant and she has been on a waiting list for several years. Her condition is deteriorating and Bob, a close friend of her family, has offered to donate her a kidney. What is the law? PMID- 11904569 TI - Improving the nutritional status of people with dementia. AB - The attention given to the nutritional needs of older adults receiving hospital care has been the subject of research over recent years. It has been suggested that older people are suffering from malnutrition as a result of poor nursing care. Those involved in care of persons with dementia are faced with considerable difficulty when trying to respond to their nutritional care needs and there is concern that there may come a time when the care team will withdraw food and fluid. However, in Southern Derbyshire we believe that we now have a set of standards for nutritional care of older adults with dementia that can ensure adequate and good nutrition despite the numerous and complex problems posed by dementia. This article discusses the progress of a sample of 20 residents of a long-stay ward over a period of 6 years and shows how a multidisciplinary team accessed, developed and applied an evidence base to practice to the benefit of the sample group. The outcomes show that malnutrition can be reversed, and that people who are considered to be in the final stages of dementia can improve their nutritional status. PMID- 11904572 TI - The accurate measurement of endotracheal tube cuff pressures. AB - The effects of endotracheal tube cuff pressure upon the tracheal wall is well documented and researched. Hyperinflation causes mucosal damage subsequent to restricted capillary blood flow (Seegobin and Hasselt, 1984), and underinflation increases aspiration risk (Bernard et al, 1979). There are critical care areas with no method of obtaining accurately cuff pressure other than adopting the minimal occlusion technique, i.e. inserting just enough air into the cuff to prevent air leakage. Although this is a useful method for obtaining an adequate seal, it does not safeguard against hyperinflation. The aim of this study was to demonstrate that without accurate measurement of intracuff pressures of endotracheal tubes, pressure will be outside the normal recommended limits, which could place patients at risk. Accurate measurements of cuff pressures post cardiac surgery were recorded using a Malincrodt pressure gauge. Theatre staff and intensive care unit nursing staff were unaware of the study until its completion. It is concluded that cuff pressures are too high using the minimal occlusion technique and the cuffs are prone to leaking. PMID- 11904571 TI - Students' experiences on neonatal nurse practitioner programmes. AB - As part of a study evaluating neonatal nurse practitioner programmes in England, data were collected on the experience of students from four separate English National Board (ENB, A19) programmes. A total of 123 nurses participated. Data were collected from 68 nurses using questionnaires at three points (the start, middle and end of the programme. The data show key concerns relating to the content and organization of course, the need for effective clinical supervision and teaching during the programmes, role transition issues and the value of informal and formal professional support. Senior and experienced nurses may find that their exposure to programmes of this kind is of interest and relevance to nurses facing similar challenges in other fields. It is also of vital importance in planning future skill mix and organizing critical care in specialties such as neonatal care. PMID- 11904573 TI - Disposable non-sterile gloves: a policy for appropriate usage. AB - Since the implementation of universal precautions, the use of disposable gloves has become part of everyday clinical practice. Guidance has been published to advise healthcare workers of the risks of latex allergies associated with the wearing of powdered latex gloves. The literature supports the use of gloves when contact with blood or body fluids is likely. However, an audit of glove usage indicated that practitioners were wearing gloves inappropriately, i.e. to wash patients (20%). The results of this study also demonstrated that a small percentage of staff (20%) were wearing vinyl gloves to deal with blood spillages and for venepuncture (13%) and cannulation (10%). A glove policy and a flow chart to assist staff in the selection of gloves were introduced. An educational programme for all hospital staff was commenced. These measures can assist healthcare workers in making an informed choice regarding glove usage. PMID- 11904574 TI - Family Nursing Network: Scottish initiative to support family care. AB - This article describes the foundation in 1997 of a Family Nursing Network (FNN) in Edinburgh involving nurses from local trusts and Scottish universities. The purpose of the FNN is to support the use of family nursing in practice, research and education. Family nursing proposes that health care should focus on the needs of the family as a unit rather than as the context of care for a family member with health needs. Family nursing is not common practice in the UK but some nurses have found it highly relevant to their care. The World Health Organization (WHO) promotes the family health nurse as an important component of health care in the community for the future and family nursing could well provide the foundation for such practice. Definitions of the family and family nursing are offered, the conceptual background is analysed and the work achieved by the FNN is reviewed. The authors write on behalf of the FNN. PMID- 11904575 TI - Biogel Super-Sensitive and Biogel Indicator glove systems. AB - Surgical gloves have been in use since the late 19th century. Initially, gloves were introduced to protect clinical staff from the carbolic acid used to sterilize equipment and prepare skin before surgery. During the mid-20th century emphasis was more towards protecting the patient from the clinician's skin flora and now, at the beginning of the 21st century, the balance lies between protecting the patient from the clinician and the clinician from the patient. The three main types of glove are latex, vinyl and polythene and they can be used singly or as part of a double gloving system. This article discusses how one hospital evaluated the glove systems Biogel Super-Sensitive and Biogel indicator underglove from Regent Medical (a division of SSL International). The results indicate that both systems are acceptable to clinical staff in a variety of specialty areas. PMID- 11904576 TI - Nurses must not suppress their inner emotions. PMID- 11904577 TI - K/DOQI clinical practice guidelines for chronic kidney disease: evaluation, classification, and stratification. PMID- 11904588 TI - The influence of luteinizing hormone-releasing hormone analog on serum leptin and body composition in women with solitary uterine myoma. AB - OBJECTIVE: The purpose of this study was to examine the influence of luteinizing hormone-releasing hormone analog goserelin on serum leptin and body composition in women with solitary uterine myoma. STUDY DESIGN: Fifteen women who were regularly menstruating and not obese were included. In all subjects, serum concentrations of leptin, insulin, testosterone, progesterone, and estradiol and body mass index and waist-to-hip ratio were measured before and after 4, 8, and 12 weeks of treatment with goserelin (3.6 mg every 4 weeks). Fat mass and lean body mass were measured by dual energy radiographic densitometry at baseline and after 12 weeks of therapy. Data were analyzed by multiple way analysis of variance and both simple and multiple regression. RESULTS: The treatment caused a significant regression of myoma. Body weight, fat, and lean mass were unchanged. No changes in plasma leptin (even after correction for fat mass) were noted during the treatment. Plasma estradiol decreased below castrate levels. Plasma progesterone decreased significantly, and testosterone tended to decline during the study. At baseline a highly significant positive correlation was found between serum leptin and fat mass. In a multiple regression analysis, neither the change in fat mass nor any of the hormonal parameters explained the significant portion of variance of plasma leptin during the treatment. CONCLUSION: Pharmacologic gonadectomy does not influence plasma leptin concentrations in women if body fat mass is unchanged. PMID- 11904589 TI - The relationship among serum insulin-like growth factor-I, insulin-like growth factor-I gene polymorphism, and bone mineral density in postmenopausal women in Korea. AB - OBJECTIVE: The objective of this study was to test the hypothesis that the cytosine-adenine polymorphism in the insulin-like growth factor-I gene is associated with serum insulin-like growth factor-I levels and bone mineral density. STUDY DESIGN: The insulin-like growth factor-I cytosine-adenine polymorphism was analyzed in 300 postmenopausal Korean women. Serum insulin-like growth factor-I, bone alkaline phosphatase, and carboxy-terminal cross-linking telopeptide of type I collagen were measured by immunoradiometric assay and enzyme-linked immunosorbent assay. Bone mineral density at the lumbar spine and proximal femur was determined by dual energy radiograph absorptiometry. RESULTS: Serum insulin-like growth factor-I and bone mineral density levels in women who were homozygous for a 194-base pair allele were significantly higher than those levels in the 194-base pair heterozygotes or women who did not possess the 194 base pair allele. A significantly decreased prevalence of the 194/194 genotype was observed in the combined group of women with osteopenia and osteoporosis, compared with normal women. No correlation between insulin-like growth factor-I genotypes and bone turnover markers was found. CONCLUSION: The insulin-like growth factor-I gene cytosine-adenine polymorphism relates with circulating insulin-like growth factor-I levels and bone mineral density at the lumbar spine and proximal femur. PMID- 11904590 TI - Age effects on urethral striated muscle. I. Changes in number and diameter of striated muscle fibers in the ventral urethra. AB - OBJECTIVE: This study was undertaken to test the null hypothesis that the number of striated muscle fibers in the ventral wall of the female urethra remains constant with increasing age. STUDY DESIGN: The urethra and surrounding tissues from 25 female cadavers, mean age 52 years (+/-SD 18, range 15-80 years), were selected for this study. Each specimen was divided along the midsagittal plane, and a Masson trichrome histologic section was prepared. A systematic count of striated muscle fibers in the ventral wall was then obtained at each decile of urethral length. RESULTS: A decrease in the total number of fibers within the sampled area was found with increasing age. The mean of the total fibers across all urethrae was 17,423 (+/-SD 9,624, range 4,788-35,867). Over the life span, an average of 364 fibers (2%) were lost per year (95% CI 197-531; P <.001). Mean fiber density was 671 (+/- SD 296, range 228-1374) fibers/mm2 and decreased by 13 fibers/mm2 per year (95% CI 8-17; P <.001). The mean lesser fiber diameter was 24 microm and did not change significantly with age ( P =.3). CONCLUSIONS: The number and density of urethral striated muscle fibers decline with age. PMID- 11904591 TI - Age effects on urethral striated muscle. II. Anatomic location of muscle loss. AB - OBJECTIVE: The purpose of the study was to measure the thickness and cross sectional area of urethral muscle layers to identify localized striated muscle loss. STUDY DESIGN: The urethra and surrounding tissues from 25 female cadavers (mean age, 52 +/- 18 [SD] years; range, 15-80 years) were used for this study. Axial and median sagittal histologic sections were prepared. Median sagittal muscle layer thickness was measured every 10% of urethral length (each decile) in the dorsal wall (adjacent to the vagina) and ventral wall, beginning at the caudal margin of the detrusor muscle (0%) and ending at the caudal margin of the striated muscle (100%). In the midurethral cross-section, the thickness of each layer was measured along radial lines placed every 45 degrees with 0 degrees at the ventral midline and 180 degrees at the dorsal midline. RESULTS: In the median sagittal sections, striated muscle layers of urethras were thinner at the vesical neck in older women. In the ventral wall, it decreased by a mean of 18 to 23 microm (3.4%-4.3%; P <.001) per year at 10% to 30% of urethral wall length. Dorsal striated muscle layers were thinner at every decile by 11 to 16 microm (3.2%-4.3%; P <.05); their total cross-sectional areas decreased by 0.19 mm2) (3.8%) per year short ( P <.001). In the midurethral cross-sections, the muscle was thinner by 16 to 25 microm (1.5% and 4.6%; P <.05) at 90, 135, and 180 degrees. CONCLUSION: Striated muscle was lost at the bladder neck and along the dorsal wall of the urethra as women aged. PMID- 11904592 TI - A deficiency in interferon-alpha production in women with vulvar vestibulitis. AB - OBJECTIVE: Previous studies have suggested that interferon-alpha may be an effective treatment for some women with vulvar vestibulitis. We evaluated whether women with this syndrome had a deficiency in endogenous and induced interferon alpha production. STUDY DESIGN: Blood was collected in heparinized tubes from 62 women with vulvar vestibulitis and 47 control women of similar age and ethnicity. Whole blood cultures were incubated in the presence of 0.1 ng/mL lipopolysaccharide (induced) or culture medium (uninduced) for 18 to 20 hours. Aliquots were tested for interferon-alpha levels by enzyme-linked immunosorbent assay. Vestibular samples were tested for human papillomavirus by polymerase chain reaction. Aliquots were also characterized for alleles of the polymorphic gene, interleukin-1 receptor antagonist, by polymerase chain reaction. RESULTS: In uninduced cultures, interferon-gamma was present in 68.1% of control subjects as opposed to 33.9% of vulvar vestibulitis patients ( P =.0005). Similarly, after lipopolysaccharide stimulation, 70.2% of control subjects and only 48.4% of patients produced interferon-alpha ( P =.03). Among the positive samples, however, there were no differences in the interferon-alpha levels between patients and control subjects. In contrast, induction of interferon gamma in response to lipopolysaccharide was similar in control subjects (78.0%) and vulvar vestibulitis patients (82.1%). Women who have a deficiency in interferon-alpha production did not have an increased prevalence of human papillomavirus infection. There was no relation between interleukin-1 receptor antagonist genotype and interferon-alpha production. CONCLUSION: An inability to produce interferon-alpha may contribute to chronic vestibular inflammation in some women. PMID- 11904593 TI - Randomized clinical trial of PCR-determined human papillomavirus detection methods: self-sampling versus clinician-directed--biologic concordance and women's preferences. AB - OBJECTIVE: The purpose of this study was to compare the high-risk human papillomavirus detection rates from self-sampled swabs and tampons with standard clinician-directed speculum sampling and to assess women's acceptance of self sampling methods. STUDY DESIGN: One hundred three women who required a colposcopy underwent order randomization of the human papillomavirus sampling technique. Kappa and McNemar test statistical results were used to measure the agreement between clinician-directed and self-sampling techniques for high-risk types of human papillomavirus and the acceptance of self-sampling techniques. RESULTS: All self-directed samplings were equivalent to clinician sampling for all cervical intraepithelial neoplasia disease states. High-risk human papillomavirus was detected by self- and clinician-directed methods in 83% of the women with cervical intraepithelial neoplasia, grade 2/3. The 2 sequential swabs trend toward better detection of high-risk types of human papillomavirus than all other techniques for women with normal histologic factors (P =.0736, by McNemar's chi2 test). Ninety-four percent of women would accept self-sampling for their yearly cervical screen. CONCLUSION: Self-sampling is equivalent to clinician sampling for the detection of high-risk human papillomavirus and is acceptable to women as a yearly screen. PMID- 11904594 TI - Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo. AB - OBJECTIVE: The objective of this study was to assess the potential of 3 spectroscopic techniques (intrinsic fluorescence, diffuse reflectance, and light scattering) individually and in combination (trimodal spectroscopy) for the detection of cervical squamous intraepithelial lesions. STUDY DESIGN: The study was conducted with 44 patients who underwent colposcopy for the evaluation of an abnormal Papanicolaou smear. Fluorescence and reflectance spectra were collected from colposcopically normal and abnormal sites and analyzed to extract quantitative information about tissue biochemistry and morphologic condition. This information was compared with histopathologic classification, and diagnostic algorithms were developed and validated with the use of logistic regression and cross-validation. RESULTS: Diagnostically significant differences exist in the composition of fluorescing biochemicals, the scattering properties, and the epithelial cell nuclear morphology of cervical squamous intraepithelial lesions and non-squamous intraepithelial lesions. Trimodal spectroscopy is a superior tool for the detection of cervical squamous intraepithelial lesions than any 1 of the techniques alone. CONCLUSION: Trimodal spectroscopy has the potential to improve the in vivo detection of precancerous cervical changes. PMID- 11904595 TI - Tubal surface lidocaine mediates pre-emptive analgesia in awake laparoscopic sterilization: a prospective, randomized clinical trial. AB - OBJECTIVE: The purpose of this study was to determine whether lidocaine that is instilled onto the Fallopian tubes reduces pain scores in awake patients who undergo laparoscopic sterilization with Filshie clips. STUDY DESIGN: This was a prospective, randomized, placebo-controlled, double-blinded, clinical trial study that was approved by our institutional review board. RESULTS: Pain scores (visual analogue scales) were lower in the lidocaine group (n = 12 patients) than in the placebo group (n = 12 patients) at clip application (6 vs 71 mm; P <.0001) and after 15 minutes after operation (15.5 vs 44.5 mm; P <.005). No significant differences occurred at 1-hour after operation or discharge, but more rescue analgesia was required in the placebo group ( P <.05), with more side effects ( P <.05). In a separate group of 20 women, serum lidocaine levels were measured (maximum level, 16.0 micromol/L). Holter monitoring of these patients revealed no significant arrhythmias. CONCLUSION: One percent lidocaine that is instilled onto the Fallopian tubes reduces pain scores in awake patients who undergo laparoscopic sterilization with Filshie clips. PMID- 11904597 TI - Managing atypical squamous cells of undetermined significance (ASCUS): human papillomavirus testing, ASCUS subtyping,or follow-up cytology? AB - OBJECTIVE: This study related morphologic subtype, human papillomavirus status, and a second cytologic examination to the follow-up biopsy-proven high-grade squamous intraepithelial lesion (HSIL; grade II or III cervical intraepithelial neoplasia) after a cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS). STUDY DESIGN: Seven hundred four liquid-based cervical cytology specimens were classified as normal, "ASCUS, favor reactive" (AFR), "ASCUS, not otherwise specified," "ASCUS, favor low-grade squamous intraepithelial lesion," "ASCUS, favor HSIL" (AFHS), low-grade squamous intraepithelial lesion, and HSIL. Human papillomavirus typing used polymerase chain reaction-restriction fragment length polymorphism analysis. A longitudinal review of the cytologic and histologic records of ASCUS cases with > or =1 follow up test or biopsy ascertained the frequency of a follow-up diagnosis of biopsy proven HSIL (grade II or III cervical intraepithelial neoplasia). RESULTS: Three hundred eighty-six cases (208 ASCUS, 68 normal, 86 with low-grade squamous intraepithelial lesions, and 24 with HSIL) were evaluated. High-risk human papillomavirus (HRHPV positive) was lowest with normal cytology (13%), highest with HSIL (71%), and was present in 29.8% of ASCUS cases, ranging from 22.2% (AFR) to 75% (AFHS). Most ASCUS tests (64%) were followed by a negative cytologic or histologic examination. Overall, 3.8% and 11% of ASCUS and HRHPV-positive ASCUS had histologic outcomes of HSIL. AFHS had the highest (25%) and AFR had the lowest (1.1%) proportion of HSIL outcomes. Sensitivity, specificity, and positive predictive values of human papillomavirus testing for biopsy-proven HSIL were 87.5%, 72.5%, and 11.3%, respectively. CONCLUSION: HSIL and AFHS are distinguished by the highest frequency of HRHPV types and higher rates of HSIL outcome. The remaining categories of ASCUS are heterogeneous with respect to human papillomavirus type and HSIL risk, and the value of subclassification of these entities is dependent on the practice. A human papillomavirus detection system based on polymerase chain reaction-restriction fragment length polymorphism identifies a smaller percentage of high-risk human papillomaviruses than mixed probe-based methods, probably because of the more precise exclusion of cross-reacting low-risk human papillomavirus. Negative HRHPV findings by either system show a markedly reduced risk of an HSIL outcome. However, the relative advantage of human papillomavirus testing over follow-up cytology will be influenced by the frequency of negative follow-up cytologic examination and sensitivity of liquid-based preparations in a given practice. PMID- 11904596 TI - Comparison of cycle control with a combined contraceptive vaginal ring and oral levonorgestrel/ethinyl estradiol. AB - OBJECTIVE: The purpose of this study was to compare cycle control and tolerability of the NuvaRing (NV Organon, Oss, The Netherlands), a novel combined contraceptive vaginal ring, with a standard combined oral contraceptive pill. STUDY DESIGN: Healthy women aged 18 to 40 years who requested contraception received either NuvaRing or a combined oral contraceptive containing 30 microg ethinyl estradiol and 150 microg levonorgestrel for 6 cycles in 3 similarly designed studies. Each cycle comprised 3 weeks of ring or pill use, followed by 1 ring- or pill-free week. RESULTS: Two hundred forty-seven women began the studies, 121 women with NuvaRing and 126 women with the combined oral contraceptive. Withdrawal bleeding occurred in virtually all cycles in both groups. In the NuvaRing groups, the incidence of irregular bleeding was < or =5% in all cycles; this was lower than the combined oral contraceptive groups (5.4% 38.8%). Furthermore, the incidence of a normal intended bleeding pattern was significantly higher in the NuvaRing groups than in the combined oral contraceptive groups (P <.01). Both contraceptives were well tolerated. CONCLUSION: NuvaRing has excellent cycle control and is well tolerated. PMID- 11904598 TI - Trends for inpatient treatment of tubal pregnancy in Maryland. AB - OBJECTIVE: The purpose of this study was to determine the burden of tubal pregnancy in Maryland in hospitalized patients and to elicit treatment trends. STUDY DESIGN: Patients who were admitted with tubal pregnancy from January 1, 1994, through March 31, 1999, were identified with the use of the Maryland Health Service Cost Review Commission discharge database. Combining this with census data, we calculated the incidence. Cases were then stratified by demographics, presentation, and surgeon volume. Outcome measures included type of medical treatment,conservative (salpingostomy or salpingotomy) or extirpative operation (salpingectomy, salpingo-oophorectomy, oophorectomy, hysterectomy), length of stay, charges, and disposition. The treatment groups were compared with the use of t tests and linear regression, and associations between demographics and type of operation were analyzed with logistic regression. RESULTS: The database included 3729 cases, which yielded an annual incidence of 5.2 per 10,000 women aged 15 to 45 years. Subjects averaged 29.6 years old and were predominantly African American(52.6%) and white (43.3%). Most of the women (67.8%) were seen in the emergency department and were treated surgically (90.7%). Conservative operation was performed in 18.1% of the women; extirpative operation was performed in 81.9% of the women. Significant predictors for extirpative operation were emergency department admission (odds ratio, 1.44; 95% CI, 1.18-1.75), increasing age (odds ratio, 1.07; 95% CI, 1.06-1.09), African American race (odds ratio, 1.87; 95% CI, 1.51-2.31), higher surgeon volume (odds ratio, 1.28; 95% CI, 1.04-1.57), and market area. Length of stay and total charges were higher for the extirpative group(P <.0001). The study lacked the power to detect differences in outcomes for other nonwhite races (5% power), laparoscopy versus laparotomy (15% power), or operating room charges (14% power). CONCLUSION: These data are limited to hospitalized patients and probably underestimate the true incidence of tubal pregnancy. Most patients underwent extirpative operation. Acuity of presentation and increasing age were appropriate predictors of this group. However, physician volume and black race were also predictors. This may be due to differences in the prevalence of disease, unmeasured clinical factors, patient and physician preferences for treatment, barriers that delayed care, or other socioeconomic factors. PMID- 11904599 TI - Accuracy of magnetic resonance imaging and transvaginal ultrasonography in the diagnosis, mapping, and measurement of uterine myomas. AB - OBJECTIVE: The purpose of this study was to evaluate and compare the accuracy of magnetic resonance imaging and transvaginal ultrasonography in myoma diagnosis, mapping, and measurement. STUDY DESIGN: This was a double-blind study of 106 consecutive premenopausal women who underwent hysterectomy for benign reasons. Myomas (total, 257) were exactly mapped by magnetic resonance imaging and transvaginal ultrasonography; in each patient, we counted correctly identified myomas with pathologic position as true value. RESULTS: The presence of myomas was detected with the same high level of precision by both methods (magnetic resonance imaging: sensitivity, 0.99; specificity, 0.86; transvaginal ultrasonography: sensitivity, 0.99; specificity, 0.91). The mean number of correctly identified myomas was significantly higher by magnetic resonance imaging than by transvaginal ultrasonography (mean difference, 0.51 +/- 1.03; P <.001), a difference that narrowed to 0.08 +/- 0.76 (P =.60) in 26 patients with 1 to 4 myomas and uterine volumes <375 mL. Magnetic resonance imaging and transvaginal ultrasonography myoma diameter measurements had equal and high accuracies in patients with 1 to 4 myomas. CONCLUSION: Transvaginal ultrasonography is as efficient as magnetic resonance imaging in detecting myoma presence, but its capacity for exact myoma mapping falls short of that of magnetic resonance imaging, especially in large (>375 mL) multiple-myoma (>4) uteri. PMID- 11904600 TI - The rhesus macaque as an animal model for pelvic organ prolapse. AB - OBJECTIVE: The purpose of this study was to characterize the pelvic floor of the rhesus macaque as an experimental model for human pelvic organ prolapse and to initiate an evaluation of the effects of estradiol and progesterone on the rhesus paravaginal attachment. STUDY DESIGN: Histologic specimens were prepared from the paravaginal attachment of 13 oophorectomized rhesus macaques. Three animals were treated with estradiol; 6 animals were treated with estradiol and progesterone, and 4 animals were untreated (hormone deprived). Immunocytochemistry was used to localize steroid receptors in the paravaginal attachment. RESULTS: Spontaneous pelvic organ prolapse was observed in rhesus macaques. The paravaginal attachment is comprised of dense collagen and elastic fibers that infiltrate the levator ani muscle. The fibroblasts of this attachment are estrogen and progesterone receptor positive, and the receptors are hormone responsive. CONCLUSION: The rhesus macaque has pelvic floor anatomy that is similar to women and makes an excellent experimental model for the study of prolapse. The rhesus paravaginal attachment is ligamentous and hormone sensitive. Its fibroblast activity may be modified by estrogen treatment in a manner similar to that reported in human pelvic connective tissue. The connective tissue of the paravaginal attachment interdigitates with the levator ani muscle cells, which suggests that this muscle plays a critical role in pelvic floor support. PMID- 11904601 TI - Perinatal outcome in women with recurrent preeclampsia compared with women who develop preeclampsia as nulliparas. AB - OBJECTIVE: To compare the rates and perinatal outcome in women who experienced preeclampsia in a previous pregnancy to those in women who developed preeclampsia as nulliparas. STUDY DESIGN: This is a secondary analysis of data from 2 separate multi-center trials of aspirin for prevention of preeclampsia. Women who had preeclampsia in a previous pregnancy (n = 598) were compared with nulliparous women (n = 2934). Outcome variables were rates of preeclampsia, preterm delivery at <37 and <35 weeks of gestation, small-for-gestational-age infant, abruptio placentae, and perinatal death. Data were compared by using chi-square analysis and Wilcoxon rank sum test. RESULTS: The rates of preeclampsia and of severe preeclampsia were significantly higher in the previous preeclamptic group as compared to the nulliparous group (17.9% vs 5.3%, P <.0001, and 7.5% vs. 2.4%, P <.0001, respectively). Women who had recurrent preeclampsia experienced more preterm deliveries before 37 and 35 weeks of gestation than nulliparous women who developed preeclampsia. In addition, among women who developed severe preeclampsia, those with recurrent preeclampsia had higher rates of preterm delivery both before 37 weeks (67% vs 33%, P =.0004) and before 35 weeks of gestation (36% vs 19%, P =.041), and higher rates of abruptio placentae (6.7% vs 1.5%) and fetal death (6.7% vs 1.4%) than did nulliparous women. CONCLUSION: Compared to nulliparous women, women with preeclampsia in a previous pregnancy had significantly higher rates of preeclampsia and adverse perinatal outcomes associated with preterm delivery as a result of preeclampsia. PMID- 11904602 TI - Umbilical cord plasma leptin is increased in preeclampsia. AB - OBJECTIVE: The objective of this study was to compare umbilical cord plasma leptin between infants of mothers who experienced preeclampsia and infants of control subjects and to study the relation between cord plasma leptin and infant obesity, as indicated by ponderal index. STUDY DESIGN: On the basis of a population of approximately 13,000 deliveries, we compared cord plasma leptin from preeclamptic (n = 256 women) and control pregnancies (n = 607 women) after taking the differences in gestational age and ponderal index into account. RESULTS: Cord plasma leptin increased strongly with gestational age, both in the preeclampsia group and the control subjects (P <.01), but at each gestational age the preeclampsia group had higher leptin levels than control subjects (P <.01). Adjustment for the higher ponderal index among control subjects (P <.05) did not alter the difference in leptin levels between the groups. CONCLUSION: We found higher levels of umbilical cord plasma leptin in infants of mothers who had preeclampsia (compared with infants of control subjects) after adjusting for differences in gestational age, gender, and infant ponderal index. PMID- 11904603 TI - How does early ultrasound scan estimation of gestational age lead to higher rates of preterm birth? AB - OBJECTIVE: Early ultrasound scanning estimation of gestational age is known to increase the reported preterm delivery rate (<37 completed weeks) compared with estimation by date of the last normal menstrual period, but it is unclear how this systematic difference arises. STUDY DESIGN: This study was a hospital-based study of 44,623 women who delivered a live-born or stillborn infant between January 1, 1978, and March 31, 1996, and who had both last normal menstrual period-based and early (usually at 16-18 weeks) ultrasound scan-based gestational age estimates. Cross-classification of the 2 estimates by completed weeks was used to examine the direction and magnitude of the differences between them and to compare the resulting classifications of preterm birth. RESULTS: The early ultrasound scan-based gestational age distribution was shifted uniformly to the left (ie, lower gestational age) relative to the last normal menstrual period gestational age distribution; the early ultrasound scan-based preterm delivery rate was 9.1%, which was 19.5% (n = 659 births) higher than the 7.6% rate by last normal menstrual period (P <.0001). The last normal menstrual period estimate exceeded the early ultrasound scan estimate far more often than the reverse, up to and including early ultrasound scan estimates of 40 weeks. No concentration of 4-week discrepancies was observed in either direction, as would be expected with random or systematic errors in recall of the last normal menstrual period. The absolute number of births at 37 to 39 weeks of gestation (by last normal menstrual period) that were reclassified as preterm (n = 1206 births) was much higher than the number of preterm births at 34 to 36 weeks of gestation that were reclassified as term (n = 581 births). The net increase of 625 preterm births (from 581 to 1206 births) that resulted from reclassification of births at 37 to 39 last normal menstrual period weeks accounted for 95% of the total 659-birth increase in early ultrasound scan-based preterm births at all last normal menstrual period gestational ages. CONCLUSION: Early ultrasound scanning reduces the gestational age estimate across the entire gestational age range; early ultrasound scan-based reclassification of gestational age results in a substantial increase in the prevalence of preterm births. Small downward reclassifications exceed upward reclassifications of similar magnitude, which is consistent with previous reports that delayed (>14 days) ovulation is more frequent than early (<14 days) ovulation. PMID- 11904604 TI - Endogenous mast cell degranulation modulates cervical contractility in the guinea pig. AB - OBJECTIVE: The purpose of this study was to investigate the effect of endogenous mast cell degranulation on the contractility of isolated cervical strips from nonpregnant and pregnant guinea pigs. STUDY DESIGN: Longitudinal cervical strips from nonpregnant and pregnant (mid and term) guinea pigs were used for isometric tension recording. Responses to the mast cell degranulating agent, compound 48/80, were compared in the absence or presence of different inhibitors and receptor antagonists. Concentration-response curves were obtained to histamine and 5-hydroxytryptamine in strips that were incubated with antagonists or solvent. RESULTS: Compound 48/80 and histamine significantly increased contractility of cervical strips in all 3 groups of animals. The inhibitor of mast cell degranulation significantly reduced responses to compound 48/80 and histamine-1 receptor antagonist reduced responses to histamine in all 3 groups. Histamine-1 receptor antagonist significantly inhibited responses to compound 48/80 in nonpregnant and mid pregnant guinea pigs. Histamine-2 receptor antagonist did not alter responses to compound 48/80 nor to histamine. The receptor antagonist 5-hydroxytryptamine-2 significantly inhibited cervical contractility that was induced by compound 48/80 in tissues from mid pregnant and term pregnant guinea pigs. Lipoxygenase inhibitor was effective in mid pregnant guinea pigs. Cyclooxygenase inhibitor, 5-hydroxytryptamine, and a combination of lipoxygenase and cyclooxygenase inhibitors had no effect on cervical contractility. CONCLUSION: The degranulation of mast cells releases histamine and other mediators that stimulate cervical contractility through histamine-1 receptors. Cervical infiltration and modulation of contractility by mast cells may play an important physiologic and/or pathologic role in the control of cervical function during pregnancy. PMID- 11904605 TI - Birth outcomes and need for hospitalization after delivery among women with multiple sclerosis. AB - OBJECTIVE: The purpose of this study was to determine whether women with multiple sclerosis who deliver singleton infants are more likely to have pregnancy or delivery complications or to have infants with low birth weight, preterm gestation, or malformations than women without multiple sclerosis and to compare their need for rehospitalization during the 2 years after delivery. STUDY DESIGN: This was a population-based cohort study that used Washington State linked birth certificate-hospital discharge records for singleton births from 1987 through 1996. Pregnancy course, birth outcomes, and need for rehospitalization within 2 years after delivery were compared for 198 women with multiple sclerosis and a comparison group of 1584 women. RESULTS: With the exception of maternal anemia, women with multiple sclerosis were no more likely to have pregnancy or delivery complications, nor were their infants more likely to be low birth weight or preterm or to have malformations. Affected women were, however, twice as likely to be rehospitalized during the 3 months after delivery. CONCLUSION: The increased risk of rehospitalization emphasizes a need for strong support systems and close monitoring during the 3 months after delivery. PMID- 11904606 TI - Prostaglandin-induced activation of uterine contractility in pregnant rats does not involve potassium channels. AB - OBJECTIVE: The uterus is a target for prostaglandins, especially at the end of gestation. Whether potassium channels are involved in the effect of prostaglandins is not clear. The aim of this study was to find out. STUDY DESIGN: Concentration-response relationships to prostaglandins (prostaglandin F2alpha, prostaglandin E2, and prostaglandin I2 [carbacyclin]; 10(-10) mol/L-10(-4) mol/L) were studied in isolated uterine rings from mid pregnancy (day 14) and late pregnancy (day 21) rats (Krebs solution, 5% CO2 in air, 37 degrees C; pH, 7.4). Rings were incubated for 30 minutes with either solvent or adenosine triphosphate sensitive potassium channel inhibitor or opener glibenclamide and levcromakalim or with calcium-sensitive potassium channel inhibitor or opener NS1619 and iberiotoxin, respectively. The changes in integral activity were compared after each concentration of the agent and were expressed as a percent of the basal integral activity. RESULTS: The increases in spontaneous contractile activity induced by prostaglandin E2 and carbacyclin, but not prostaglandin F2alpha, were statistically significantly higher in tissues from late pregnancy versus mid pregnancy rats and were not affected by any of the K-channel openers or inhibitors. CONCLUSION: Adenosine triphosphate-sensitive and calcium-sensitive potassium channels are not involved in the effect of prostaglandin F2alpha, prostaglandin E2, and prostaglandin I2 on pregnant rat uterus. PMID- 11904607 TI - The preterm prediction study: elevated cervical ferritin levels at 22 to 24 weeks of gestation are associated with spontaneous preterm delivery in asymptomatic women. AB - OBJECTIVE: Low serum ferritin levels correlate with low iron stores, whereas high levels are associated with an acute-phase reaction. Our objective was to determine whether elevated levels of ferritin in the genital tract may be a potent marker to identify patients at risk for spontaneous preterm delivery. STUDY DESIGN: We performed a nested case-control study involving 182 women who had spontaneous preterm delivery and 182 term control subjects matched for race, parity, and recruitment center, and selected from 2929 women enrolled in the Preterm Prediction Study of the National Institute of Child Health and Development Maternal-Fetal Medicine Units Network. Cervical fluid ferritin was measured by use of radioimmunoassay. RESULTS: Cervical ferritin levels were significantly higher in women who subsequently had spontaneous early preterm delivery (<32 weeks, mean +/- SD, 37.7 +/- 31.1 vs 21.5 +/- 24.1 ng/mL, P =.002; and <35 weeks, 43.2 +/- 62.7 vs 28.2 +/- 36.7 ng/mL, P =.004) than in term controls. A cervical ferritin of >75th percentile in the controls (>35.5 ng/mL) was found in 52.9% (9/17) of the women delivered <29 weeks vs 17.7% (3/17) of the controls (odds ratio [OR] 5.3 [95% CI 1.1-25.2]) and in 43.5% (20/46) of the women delivered <32 weeks versus 10.9% (5/46) of the controls (OR 6.3, 95% CI 2.1 18.9). Cervical ferritin levels had a weaker association with spontaneous preterm delivery <35 weeks (OR 2.8 [95% CI 1.5-5.1]) and <37 weeks (OR 1.6, 95% CI 1.0 2.5]). Cervical ferritin levels correlated significantly with cervical lactoferrin, interleukin-6 (IL-6), and defensin levels. CONCLUSIONS: Elevated cervical ferritin levels at 22 to 24 weeks of gestation in asymptomatic women are associated with subsequent spontaneous preterm birth. The strong correlation of cervical ferritin with other inflammatory markers provides support for the hypothesis of infection as a mediator of preterm delivery. PMID- 11904608 TI - Follistatin-free activin A is not associated with preterm birth. AB - OBJECTIVE: The purpose of this study was to determine whether follistatin-free activin A (an inhibin-related protein with rising serum levels before term labor) is associated with spontaneous preterm birth in an outpatient population. STUDY DESIGN: From 10 centers, 2929 women were enrolled in the Preterm Prediction Study. Plasma was isolated from blood samples obtained at 24 and 28 weeks of gestation and stored. A nested case-controlled study was performed; there were 197 women with spontaneous preterm birth before 37 weeks of gestation with 24 week samples and 142 cases with 28-week samples that were matched to an equal number of term control samples from each time point. A follistatin-free activin A enzyme-linked immunosorbent assay was performed, with results interpreted as positive or negative (defined as a value less than the assay limit of sensitivity). A comparison of follistatin-free activin A with other tests that were associated with spontaneous preterm birth (alpha-fetoprotein, alkaline phosphatase, cortisol, granulocyte colony-stimulating factor, fetal fibronectin, and cervical interleukin-6) was also performed. RESULTS: The odds ratio for associated spontaneous preterm birth at <37 weeks of gestation if the follistatin free activin A result was positive was 1.16 at 24 weeks (95% CI, 0.62-2.17) and 1.39 at 28 weeks (95% CI, 0.68-2.84). A positive follistatin-free activin A result at either 24 or 28 weeks was not associated with a positive test for any of the other laboratory tests, except granulocyte colony-stimulating factor at 28 weeks' gestation. CONCLUSION: Follistatin-free activin A is not associated with spontaneous preterm birth in an asymptomatic outpatient population. Follistatin free activin A may not be an appropriate screening marker for spontaneous preterm birth in the office setting. PMID- 11904609 TI - The optimization of intravaginal misoprostol dosing schedules in second-trimester pregnancy termination. AB - OBJECTIVE: The purpose of this study was to compare the clinical efficacy and side effects of 3 doses of intravaginal misoprostol for second-trimester pregnancy termination. STUDY DESIGN: This was a prospective randomized, double blind controlled clinical trial of 150 women who underwent pregnancy termination between 14 and 30 weeks of gestation. Three intravaginal misoprostol regimens were compared: 200 microg misoprostol at 6-hour intervals (group 1), 400 microg misoprostol at 6-hour intervals (group 2), and a loading dose of 600 microg misoprostol followed by 200 microg at 6-hour intervals (group 3). RESULTS: There was a significant difference in the median time to achieve delivery among the 3 groups: group 1 (18.2 hours [IQ, 13.3-32.5 hours]) vs group 2 (15.1 hours [IQ, 10.9-23.7 hours]) vs group 3 (13.2 hours [IQ, 11.2-21.7 hours]; P =.035). Fifty nine percent of the women in group 1, 76% of the women in group 2, and 80% of the women in group 3 delivered within 24 hours (P =.013). There were 7.8% of the women in group 1, 0% of the women in group 2, and 2% of the women in group 3 who were undelivered at 48 hours (P =.02). There was an increase in the incidence of fever in the first 12 hours (P =.038) and in the incidence of vomiting within 3 hours of the initial dose (P =.048) in group 3 compared with the other groups. CONCLUSION: Intravaginal misoprostol 400 microg at 6-hour intervals appears to be the preferred regimen for second-trimester pregnancy termination, with a shorter commencement to delivery interval than the 200 microg regimen and fewer maternal side-effects than the 600 microg loading dose regimen. PMID- 11904610 TI - Maternal benefit of corticosteroid therapy in patients with HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome: impact on the rate of regional anesthesia. AB - OBJECTIVE: Our purpose was to assess the impact of glucocorticoid administration on the rate of regional anesthesia in women with hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome. STUDY DESIGN: Maternal records of pregnancies with HELLP syndrome managed between April 1994 and December 1999 were analyzed retrospectively. RESULTS: Sixty-nine patients were identified with antepartum HELLP syndrome and 46 (66%) received glucocorticoids. The presence of thrombocytopenia at admission and the interval from presentation to delivery was evaluated to assess the impact of glucocorticoid use. In the 37 women who had platelet counts of <90,000/mm3, 0 in the untreated group (0 of 11) versus 42% in the steroid group (11 of 26) received regional anesthetic, P =.015. Furthermore, the rate of regional anesthesia increased from 0 in the untreated group delivered within 24 hours (n = 10) to 57% (8 of 14) in the glucocorticoid group, in which women attained a 24-hour latency from presentation to delivery, P =.006. The need for general anesthesia also decreased significantly in treated women who attained a 24-hour latency compared to untreated women who did not, 100% (n = 7) versus 22% (n = 9), P =.003. CONCLUSIONS: Administration of glucocorticoids increases the use of regional anesthesia in women with antepartum HELLP syndrome who have thrombocytopenia, particularly in those who achieve a latency of 24 hours before delivery. PMID- 11904611 TI - What are pregnant women eating? Nutrient and food group differences by race. AB - OBJECTIVE: The purpose of this study was to identify foods that contributed most to nutrient and fiber intake in a sample of pregnant women in North Carolina. STUDY DESIGN: This was a prospective study of women in the Pregnancy, Infection, and Nutrition Study (n = 2247 women). Dietary information during the second trimester was collected with the use of a food frequency questionnaire. The contribution of each food item to the population's intake was calculated. RESULTS: Overall, low nutrient-dense foods were major contributors to energy, fat, and carbohydrates, whereas fortified foods were important sources of iron, folate, and vitamin C. The median energy intake for this population was 2478 kcal. The median dietary intakes of iron were below the recommended levels. Although black women consumed more calories on average, white women, after energy adjustment, consumed greater amounts of protein, iron, folate, and fiber. CONCLUSION: These data emphasize the importance of evaluating both the nutrient density in the diet and the frequency of consumption in the assessment of the diets of pregnant women. PMID- 11904612 TI - The Maternal Lifestyle Study: drug exposure during pregnancy and short-term maternal outcomes. AB - OBJECTIVE: Reports of maternal effects resulting from drug exposure during pregnancy are inconsistent. The Maternal Lifestyle Study (MLS) is a multicenter, prospective, observational study that was initiated to better define the effects of exposure to illicit drugs during pregnancy on the mother, fetus, and infant. METHODS: Between May 1993 and May 1995, of 19,079 mother-infant dyads that were screened after delivery for cocaine and opiate exposure at four clinical centers (Brown University, University of Miami, University of Tennessee, Memphis, and Wayne State University), 16,988 (89%) met eligibility criteria and 11,811 (70%) of those eligible agreed to participate in the study. Exposure was defined as an admission of use of cocaine or opiates or both or the presence of cocaine or opiate metabolites in meconium as determined by use of gas chromatography-mass spectroscopy assay. Nonexposure was defined as a negative drug use history by interview and a negative immunoassay screen. When exposure could not be confirmed, such as when meconium was not obtained or was inadequate for confirmatory analysis, the mother-infant dyad was excluded (n = 3184). RESULTS: Of the mothers who consented to participate, 50% were African American, 38% were married, 64% were Medicaid recipients, and 95% had at least one prenatal care visit (median, 10 visits). Significant differences (P <.01) between cocaine opiate exposed (n = 1185) and nonexposed (n = 7442) mothers included race (African American: 74.6% and 47.0%, respectively), mean age (29.6 and 26.1 years, respectively), and polydrug use including any combination of alcohol, tobacco, and/or marijuana (93% and 42%, respectively). Odds ratios (99% CI) indicate that exposed mothers had a significantly higher risk(P <.001) of medical complications including syphilis 6.7 (4.8-9.6), gonorrhea 1.9 (1.3-3.0), and hepatitis 4.8 (2.6 8.91); psychiatric, nervous, and emotional disorders 4.0 (2.2-7.4); and abruptio placenta 2.3 (1.4-3.9). The odds of a positive test for human immunodeficiency virus were higher (available on 28% of the cohort) in the exposed group 8.2 (14.3 15.4). Seventeen cases of maternal acquired immunodeficiency syndrome (AIDS) were identified. Opiate exposure with its attendant needle use significantly increased the risk of hepatitis and AIDS. The number of hospitalizations during pregnancy did not differ between the exposure groups because 11% of patients in each group were hospitalized at least once. However, violence as a cause of hospitalization was more common in the cocaine-exposed group, 19.6 (2.7-144.7). CONCLUSION: This observational study confirmed many of the reported adverse social and serious medical perinatal complications of mothers exposed to cocaine or opiates during pregnancy. The overall prevalence of these risk outcomes was lower than has been reported previously. PMID- 11904613 TI - Oxytocin preparation stability in several common obstetric intravenous solutions. AB - OBJECTIVE: The purpose of this study was to determine the stability of oxytocin in lactated Ringer's solution and lactated Ringer's-dextrose 5% solution over a 24-hour period at 25 degrees C and over a 7-day period at 5 degrees C. STUDY DESIGN: Twenty units (2.1 microg equal 1 unit) of oxytocin were injected into 1000 mL of lactated Ringer's solution and lactated Ringer's-dextrose 5% solution. Samples for the analysis were drawn at specified times after storage at 5 degrees C and 25 degrees C. These samples were stored at -70 degrees C for later analysis. Statistical analysis was done with 1-way analysis of variance and Tukey Kramer multiple comparisons. RESULTS: Twenty units of oxytocin in 1000 mL of lactated Ringer's solution and lactated Ringer's-dextrose 5% solution was found to be stable for 7 days at 5 degrees C and for 24 hours at 25 degrees C. CONCLUSION: Premixed oxytocin solutions in lactated Ringer's solution and lactated Ringer's-dextrose 5% solution are stable in conditions commonly found in dispensing pharmacies and labor and delivery units. This finding could lead to the more efficient use of personnel during the mixing process, could provide solutions that are aseptically prepared, and could be a tool to reduce costs and improve patient care. PMID- 11904615 TI - Connexin 43 expression in normal versus dysfunctional labor. AB - OBJECTIVE: We sought to determine whether connexin 43 (Cx43), the major myometrial gap junction protein, is differentially expressed in normal versus dysfunctional labor. STUDY DESIGN: Myometrial biopsies were obtained from 28 patients undergoing cesarean section and grouped into the following categories: (1) no labor, (2) dysfunctional labor, and (3) normal labor. Northern and Western analyses were performed to determine Cx43 messenger RNA (mRNA) and protein expression, respectively. Localization of Cx43 protein was determined by immunohistochemistry. RESULTS: Labor was associated with increased Cx43 mRNA expression (P <.05). This association was not true for protein. There was no difference in mRNA or protein expression between patients with normal labor versus those with dysfunctional labor. The extent of Cx43 immunohistochemical staining was not significantly different among the groups (P >.05). CONCLUSION: Dysfunctional labor is not associated with aberrant Cx43 mRNA or protein expression or with a reduction in immunodetectable Cx43 gap junctions. PMID- 11904614 TI - Maternal and fetal plasma homocysteine concentrations at birth: the influence of folate, vitamin B12, and the 5,10-methylenetetrahydrofolate reductase 677C-->T variant. AB - OBJECTIVE: The purpose of this study was to determine the nutritional and genetic factors that influence fetal plasma homocysteine concentrations. STUDY DESIGN: Maternal and umbilical cord venous blood was taken from 201 women who were delivered after uncomplicated pregnancies of 37 to 41 gestational weeks. Red blood cell folate, plasma folate, plasma vitamin B12, and plasma homocysteine concentrations were measured in all samples. Cord and maternal bloods were also genotyped for the 677C-->T variant. RESULTS: The fetal circulation had lower homocysteine concentrations (7.87 +/- 2.87 micromol/L; mean +/- SD) than the maternal circulation (8.34 +/- 2.94 micromol/L; P =.003), but there was a strong linear association between the levels in these 2 compartments (r = 0.72; P <.0001). Overall, the maternal homocysteine level had the strongest influence on the fetal homocysteine concentration (P <.0001), with the fetal and maternal vitamin B12 having important additional effects (P =.016 and P =.0045, respectively). Fetal plasma folate and 5,10-methylenetetrahydrofolate reductase 677C-->T genotype had no significant effects (P =.23 and P =.54, respectively), probably because of folic acid supplement use. CONCLUSION: The maternal homocysteine level is the primary predictor of blood homocysteine in the developing fetus. If lowering maternal blood homocysteine proves critical to preventing pregnancy complications, it will be important to maintain optimal vitamin B12 status in addition to optimal tissue folate status. PMID- 11904616 TI - Human placental growth hormone causes severe insulin resistance in transgenic mice. AB - OBJECTIVE: The insulin resistance of pregnancy is considered to be mediated by human placental lactogen, but the metabolic effects of human placental growth hormone have not been well defined. Our aim was to evaluate the effect of placental growth hormone on insulin sensitivity in vivo using transgenic mice that overexpress the human placental growth hormone gene. STUDY DESIGN: Glucose and insulin tolerance tests were performed on 5 transgenic mice that overexpressed the human placental growth hormone variant gene and 6 normal littermate controls. The body composition of the mice was assessed by dual-energy radiograph absorptiometry, and free fatty acid levels were measured as a marker of lipolysis. RESULTS: The human placental growth hormone levels in the transgenic mice were comparable to those attained in the third trimester of pregnancy. These mice were nearly twice as heavy as the control mice, and their body composition differed by a significant increase in bone density and a small decrease in percentage of body fat. Fasting insulin levels in the transgenic mice that overexpressed placental growth hormone were approximately 4-fold higher than the control mice (1.57 +/- 0.22 ng/mL vs 0.38 +/- 0.07 ng/mL; P <.001) and 7 times higher 30 minutes after glucose stimulation (4.17 +/- 0.54 ng/mL vs 0.62 +/ 0.10 ng/mL; P <.0001) with no significant difference in either fasting or postchallenge glucose levels. Insulin sensitivity was markedly decreased in the transgenic mice, as demonstrated by an insignificant decline in glucose levels after insulin injection compared with the control mice, which demonstrated more than a 65% reduction in glucose levels (P <.001). CONCLUSION: Human placental growth hormone causes insulin resistance as manifested by fasting and postprandial hyperinsulinemia and minimal glucose lowering in response to insulin injection. Human placental growth hormone is a highly likely candidate to mediate the insulin resistance of pregnancy. PMID- 11904617 TI - Iron therapy in iron deficiency anemia in pregnancy: intravenous route versus oral route. AB - OBJECTIVE: The aim of this study was to compare intravenous iron sucrose versus oral iron sulfate in anemia at 6 months of pregnancy. STUDY DESIGN: A random, prospective, open study with individual benefit was performed involving 50 patients with hemoglobin levels between 8 and 10 g/dL and a ferritin value of <50 microg/L. In the intravenous group (IV group), the iron dose was calculated from the following formula: Weight before pregnancy (kg) x (120 g/L - Actual hemoglobin [g/L]) x 0.24 + 500 mg. The oral group (PO group) received 240 mg of iron sulfate per day for 4 weeks. Treatment efficacy was assessed by measurement of hemoglobin and reticulocytes on days 8, 15, 21, and 30 and at delivery and of ferritin on day 30 and at delivery. The baby's birth weight and iron stores were noted. Results were expressed as median +/- interquartile range. Mann-Whitney and Wilcoxon tests were used for the analysis, with P <.05 considered significant. RESULTS: An increase in hemoglobin was observed, rising from 9.6 +/- 0.79 g/dL to 11.11 +/- 1.3 g/dL on day 30 in the IV group and from 9.7 +/- 0.5 g/dL to 11 +/- 1.25 g/dL on day 30 in the PO group (not significant). On day 30 (P <.0001) and at delivery (P =.01) ferritin was higher in the IV group. A mean higher birth weight of 250 g was noted in the IV group (not significant). CONCLUSION: Iron sucrose appears to be a treatment without serious side effects indicated in correction of pregnancy anemia or iron stores depletion. PMID- 11904618 TI - Bacterially induced preterm labor in the mouse does not require maternal interleukin-1 signaling. AB - OBJECTIVE: We tested the hypothesis that intrauterine bacterial inoculation induces labor via expression of the inflammatory cytokine interleukin-1 (IL-1) in a murine model. STUDY DESIGN: Pregnant mice on day 14.5 of a 19-20 day gestation were inoculated with killed Escherichia coli or sterile media into either (a) the right uterine horn, (b) the right uterine horn following its surgical isolation from the contralateral horn and cervix, or (c) the kidney. Cytokine levels in gestational tissues and maternal serum were determined by use of enzyme-linked immunosorbent assay (ELISA). In a separate experiment, bacterially induced preterm delivery was compared between mice lacking a functional IL-1 receptor and wild-type control litter mates. RESULTS: Killed E coli induced delivery within 48 hours with similar dose-response curves regardless of inoculation site (intact uterine horn, isolated uterine horn, or kidney). Bacterial inoculation of an isolated right horn caused dramatic increases in local expression of IL-1 and IL 6. However, delivery occurred from the uninjected horn without corresponding upregulation of cytokines, with the exception of a modest rise within fetal membranes. Mice lacking a functional IL-1 receptor were no different from wild type mice in their susceptibility to bacterially induced delivery. CONCLUSION: Bacterially induced labor in the murine model does not require IL-1 signaling. PMID- 11904619 TI - Stereologic examination of placentas from mothers who smoke during pregnancy. AB - OBJECTIVE: Structural changes in the placenta might contribute to the lower birth weight seen among infants born to mothers who smoke cigarettes. In this study, a morphologic examination and a stereologic quantitation of placentas from mothers who smoked cigarettes and who did not smoke cigarettes during pregnancy were performed. STUDY DESIGN: Twenty-five placentas from mothers who did not smoke cigarettes, 15 placentas from mothers who smoked 5 to 10 cigarettes per day, 16 placentas from mothers who smoked 11 to 20 cigarettes per day, and 16 placentas from mothers who smoked >20 cigarettes per day were delivered at term after normal pregnancies and were fixed by dual perfusion. The volume and the surface area of villi, the trophoblast volume, and the volume and the surface area and length of villous capillaries were estimated. A measurement of the concentration of cadmium in serum was used to assess the validity of information concerning smoking habits. RESULTS: No differences were shown in the total volume of placenta between the groups. The estimated volume and surface area and the calculated lengths for villous capillaries were significantly reduced in all 3 groups of smokers. A significant increase of the trophoblast volume was observed in the mothers who smoked cigarettes. CONCLUSION: Cigarette smoking during pregnancy influences the placental vasculature. The reduced dimensions of fetal capillaries in villi may affect the placental blood flow, and the diminished area for exchange of gases and nutrients between the mother and the fetus will increase the risk of fetal undernourishment. PMID- 11904620 TI - Nitrous oxide amniodistention compared with fluid amniodistention reduces operation time while inducing no changes in fetal acid-base status in a sheep model for endoscopic fetal surgery. AB - OBJECTIVE: Our purpose was to evaluate in a sheep model for endoscopic fetal surgery the impact of nitrous oxide-gas amniodistention compared with fluid amniodistention on duration of surgery, hemorrhagic events, and fetomaternal acid base status. STUDY DESIGN: Pregnant ewes (n = 16) at 92 to 104 days of gestation (term, 145 days) underwent amniodistention with Hartmann's solution (group I, n = 8) or nitrous oxide (group II, n = 8) at 38 degrees C. Endoscopic tracheal clipping according to a standardized surgical protocol was performed in all animals. The duration of fetoscopy (from insertion of first cannula until removal of last one), fetal surgery (fetal skin incision to skin closure), and number of bleeding episodes was recorded. Maternal and fetal blood gas values (pH, PO2, and PCO2) were measured at baseline and every 15 minutes during the experiment. Videotapes of the operations were assessed independently and scored by a visual analog scale in terms of smoothness of the operation and control of hemorrhagic events. RESULTS: Mean duration of fetoscopy (+/-SEM) (68 +/- 16 minutes vs 92 +/- 23 minutes) and fetal surgery (19 +/- 6 minutes vs 42 +/- 18 minutes) as well as number of bleeding episodes (1.9 +/- 0.8 minutes vs 5.8 +/- 2.1 minutes) was significantly reduced in animals operated with use of nitrous oxide amniodistention. In both groups, fetal and maternal blood gases remained unchanged during the entire experiment. Visual analog scale (VAS) scores were significantly higher for procedures conducted with use of gas distention. CONCLUSION: In a lamb model for fetal surgery, gas amniodistention with use of nitrous oxide results in a quicker operation procedure with less bleeding compared with fluid amniodistention, and without adverse side effects on fetomaternal acid-base status. PMID- 11904621 TI - Stress in pregnancy: a new Wistar rat model for human preeclampsia. AB - OBJECTIVE: Our study evaluated the effects of chronic and/or acute stress on pregnant and nonpregnant female rats. STUDY DESIGN: The rats were exposed to the sonic stimulus associated with overpopulation between days 7 and 14 of pregnancy. The rats were immobilized 2 days before the vascular reactivity experiments. RESULTS: In 14-day pregnant rats, chronic stress led to lower weight, increased adrenal weight, lower endothelium-derived relaxing factor release, and lower fetal weight. In 20-day pregnant rats, chronic stress caused decreased weight gain, higher blood pressure, increased vasomotility and proteinuria, lower endothelium-derived relaxing factor release, and lower fetal weight. In the 20 day pregnant group, the higher adrenal weight resulted in higher blood pressure, lower vascular relaxation, and lower average fetal weight. A greater number of fetuses had higher adrenal weight, higher blood pressure, and lower vascular relaxation. CONCLUSION: The alterations found in the rats were similar to those that occur in human preeclampsia. Therefore, we propose a new animal model for human preeclampsia. PMID- 11904622 TI - Effects of nicotine on intercellular adhesion molecule expression in endothelial cells and integrin expression in neutrophils in vitro. AB - OBJECTIVE: We previously reported that nicotine decreases leukocyte adhesion to uterine vascular endothelial cells in vivo under ischemic conditions in pregnant rabbits. To further investigate the mechanism of decreased leukocyte-endothelial adhesion by nicotine exposure, the effect of nicotine on endothelial cell intercellular adhesion molecule expression and neutrophil integrin expression of CD62L, CD11a, and CD11b were examined. STUDY DESIGN: Endothelial cells were isolated from human umbilical cord veins from normal pregnancies in nonsmoking women immediately after delivery. Neutrophils were isolated from healthy nonpregnant and nonsmoking female volunteers. First passage of endothelial cells and fresh isolated neutrophils were exposed to nicotine at different concentrations. Surface adhesion molecule expression of intercellular adhesion molecule on endothelial cells was determined by colorimetric assay. Neutrophil integrin expressions for CD62L, CD11a, and CD11b were determined by flow cytometry. Messenger RNA expression for intercellular adhesion molecule in endothelial cells was examined by reverse transcription-polymerase chain reaction. RESULTS: Nicotine at a lower concentration of 0.01 micromol/L had no effect on endothelial cell surface intercellular adhesion molecule expression compared with controls (P =.614). Nicotine at a higher concentration of 10 micromol/L completely inhibited endothelial cell surface intercellular adhesion molecule-1 expression(P <.0001). At concentrations between 0.10 and 10 micromol/L, nicotine inhibited intercellular adhesion molecule expression in a dose-dependent manner. Messenger RNA expression of intercellular adhesion molecule in endothelial cells was not changed after exposure to nicotine. Decreased integrin expressions of CD62L, CD11a, and CD11b were observed on neutrophils after exposure to nicotine. CONCLUSION: Nicotine exerts inhibitory effects on both endothelial cell surface intercellular adhesion molecule expression and neutrophil integrin expressions of CD62L, CD11a, and CD11b in vitro. These in vitro effects of nicotine may relate to the clinical observation of reduced incidence of preeclampsia in women that smoke. PMID- 11904623 TI - Endothelial cell apoptosis is induced by fetal plasma from pregnancy with umbilical placental vascular disease. AB - OBJECTIVE: Vascular disease in the umbilical placental circulation that is detected by umbilical artery Doppler study is associated with adverse fetal outcome. Endothelial cell activation and platelet consumption are features of this pathologic condition. We postulated that this was due to the local release of factors that cause endothelial cell injury and that these would spill into the fetal circulation. To test this hypothesis, we examined for the presence in fetal plasma of factors that induced endothelial cell apoptosis in pregnancies that were complicated by umbilical placental vascular disease. STUDY DESIGN: Isolated and cultured human umbilical vein endothelial cells were exposed to fetal plasma from the 3 fetal groups: normal pregnancy (n = 32 patients), pregnancy with umbilical placental vascular disease that was identified by an abnormal umbilical artery Doppler study (n = 38 patients), and pregnancy with maternal preeclampsia and normal umbilical artery Doppler study (n = 16 patients). Early apoptosis can be recognized by a loss of plasma membrane asymmetry with membrane uptake of annexin V. This was measured with annexin V and propidium iodide staining by fluorescent-activated cell scanning. Cells that underwent early apoptosis stained positive for annexin V and negative for propidium iodide (in contrast with cells that underwent necrosis). Cytosolic proteolytic activity was also measured. The lysates from endothelial cells that were stimulated by fetal plasma from umbilical placental vascular disease were tested for caspase-3 and caspase-8 activities by a fluorescent assay with spectrofluorophotometry. RESULTS: The percentage of endothelial cells that underwent apoptosis was significantly higher (P <.05) when stimulated with fetal plasma from pregnancies with umbilical placental vascular disease (17.71% +/- 1.31%) than with fetal plasma from normal pregnancies (9.76% +/- 0.87%). In the presence of maternal preeclampsia with normal umbilical artery Doppler study, the percent of apoptotic cells (11.31% +/- 1.59%) was similar to that of the normal group. In the group with abnormal umbilical artery Doppler study, there was no difference between pregnancies with preeclampsia (n = 17 pregnancies) and without preeclampsia (n = 21 pregnancies). The protease activity of caspase-3 was significantly enhanced in the group with umbilical placental vascular disease compared with normal pregnancy (0.79 +/- 0.06 vs 0.45 +/- 0.08 microMol/L). However, no difference in caspase-8 activity was detected (0.66 +/- 0.05 vs 0.56 +/- 0.05 microMol/L). CONCLUSION: Endothelial cell apoptosis is a feature of umbilical placental vascular disease. Our study demonstrates the presence of factors in the fetal plasma that caused endothelial cells to undergo early apoptosis. This increased apoptosis was only seen in the presence of placental vascular disease and was independent of the presence or absence of maternal preeclampsia. Our results indicate that programmed endothelial cell death occurs in the fetal circulation as a part of the injury that is associated with the development of umbilical placental vascular disease. The caspase-3, rather than caspase-8, signal transduction pathway appears to be involved in the mediation of endothelial cell apoptosis that was detected in our study. PMID- 11904624 TI - An evaluation of the effects of sucrose on neonatal pain with 2 commonly used circumcision methods. AB - OBJECTIVE: The purpose of this study was to identify the least painful circumcision method. STUDY DESIGN: The infants were circumcised with either the Mogen or the Gomco procedure and were given a sweetened pacifier or a pacifier dipped in water. All infants had a eutectic mixture of local anesthetic cream applied before circumcision. The duration of the crying and grimacing were measured. RESULTS: The Gomco procedure took 1.9 times longer to complete. Infants who were circumcised with the Mogen procedure cried and grimaced far less than infants who were circumcised with the Gomco procedure (P =.0001). Sucrose on a pacifier was far more analgesic than water on a pacifier for infants in the Gomco group. CONCLUSION: On the basis of these and other findings on pain prevention and amelioration, we recommend that a local anesthetic be administered in advance of circumcision and that the Mogen procedure be used, unless contraindicated. We also recommend that infants be given a sweetened pacifier before, during, and after circumcision if the Gomco method is used. PMID- 11904625 TI - Congenital syphilis after maternal treatment for syphilis during pregnancy. AB - OBJECTIVE: The purpose of this study was to characterize pregnancies that were complicated by maternal syphilis that had been treated before delivery in which the newborn infant was diagnosed with congenital syphilis. STUDY DESIGN: Prospective surveillance from January 1, 1982, to December 31, 1998, involved women who received antenatal treatment for syphilis. Infants who were born with congenital syphilis were identified by clinical or laboratory criteria. Antepartum factors such as gestational age, time to delivery and VDRL titers were then analyzed and compared with those of women who had been treated and who were delivered of an uninfected infant. The 1:1 match was based on the stage of syphilis and the gestational age at treatment. RESULTS: Forty-three women who received antepartum therapy for syphilis were delivered of an infant with congenital syphilis. Most of the women had been treated for early syphilis; the mean gestational age at treatment was 30.3 weeks. Thirty-five percent of the women were treated >30 days before delivery. Fifty-six percent of the infants were preterm. The 1:1 match revealed that treatment and delivery high VDRL titers, prematurity, and a short interval from treatment to delivery were significantly different in those infants who were diagnosed with congenital syphilis. CONCLUSION: High VDRL titers at treatment and delivery, earlier maternal stage of syphilis, the interval from treatment to delivery, and delivery of an infant at < or =36 weeks' gestation are associated with the delivery of a congenitally infected neonate after adequate treatment for maternal syphilis. PMID- 11904626 TI - Effect of dexamethasone on mitochondrial maturation in the fetal rat brain. AB - OBJECTIVE: The objective of the present study was to explore whether prenatal dexamethasone treatment influences mitochondrial maturation in the fetal rat brain. STUDY DESIGN: Mitochondrial respiration was measured polarographically with homogenates of fetal cerebral cortical tissues on day 16 (with saline solution, n = 8; with dexamethasone, n = 8), day 18 (with saline solution, n = 8; with dexamethasone, n = 8), and day 20 (with saline solution, n = 8; with dexamethasone,n = 8) of gestation. Four doses of dexamethasone (0.1 mg small middle dot kg) or vehicle (saline solution) were given, with an interval of 12 hours, until 12 hours before each measurement. RESULTS: In the vehicle-treated animals, mitochondrial respiratory activity was increased significantly after day 18 of gestation. Dexamethasone-treated animals showed a significant increase in mitochondrial activity at day 16 of gestation compared with vehicle-treated animals. CONCLUSION: The results indicate that prenatal dexamethasone treatment contributes to the precocious maturation of mitochondrial activity in the fetal rat brain. Because acceleration in cerebral mitochondrial activities is required immediately after birth to maintain high-energy phosphate levels, the precocious maturation may be crucial for the successful outcome of the preterm infant. PMID- 11904627 TI - Does chorionicity or zygosity predict adverse perinatal outcomes in twins? AB - OBJECTIVE: The purpose of this study was to evaluate chorionicity and zygosity as risk factors for adverse perinatal outcomes in twins. STUDY DESIGN: A population based, retrospective cohort study was conducted of all twin deliveries in Nova Scotia, Canada, from 1988 to 1997. Chorionicity was established by histologic examination. Zygosity was determined by chorionicity, sex, and infant blood group. Three groups were established: monochorionic/monozygotic twins, dichorionic/dizygotic twins, and dichorionic/majority monozygotic twins. RESULTS: Outcomes from 1008 twin pregnancies were analyzed. Monochorionic/monozygotic twins had lower mean birth weights compared with dichorionic/dizygotic twins. Rates of perinatal mortality of at least 1 twin were significantly higher among monochorionic/monozygotic twins relative to dichorionic/dizygotic twins (relative risk, 2.5; 95% CI, 1.1-2.5). Dichorionic/majority monozygotic twins had similar perinatal outcomes compared with dichorionic/dizygotic twins. CONCLUSION: Monochorionicity increases the risk of adverse perinatal outcome, whereas the effect of zygosity is less clear. Because chorionicity can be determined by prenatal ultrasound scanning, this information should be considered in the prenatal care of twin pregnancies. PMID- 11904628 TI - Statistics usage in the American Journal of Obstetrics and Gynecology: has anything changed? AB - OBJECTIVE: Our purpose was to compare statistical listing and usage between articles published in the American Journal of Obstetrics and Gynecology in 1994 with those published in 1999. STUDY DESIGN: All papers included in the obstetrics, fetus-placenta-newborn, and gynecology sections and the transactions of societies sections of the January through June 1999 issues of the American Journal of Obstetrics and Gynecology (volume 180, numbers 1 to 6) were reviewed for statistical usage. Each paper was given a rating for the cataloging of applied statistics and a rating for the appropriateness of statistical usage, when possible. These results were compared with the data collected on a similar review of articles published in 1994. RESULTS: Of the 238 available articles, 195 contained statistics and were reviewed. In comparison to the articles published in 1994, there were significantly more articles that completely cataloged applied statistics (74.3% vs 47.4%) (P <.0001), and there was a significant improvement in appropriateness of statistical usage (56.4% vs 30.3%) (P <.0001). CONCLUSION: Changes in the Instructions to Authors regarding the description of applied statistics and probable changes in the behavior of researchers and Editors have led to an improvement in the quality of statistics in papers published in the American Journal of Obstetrics and Gynecology. PMID- 11904629 TI - Antibiotic therapy for the treatment of preterm labor: a review of the evidence. AB - OBJECTIVE: The purpose of this study was to evaluate the evidence regarding antibiotics for the treatment of preterm labor. STUDY DESIGN: Through dual review, we abstracted study design and masking, definitions of preterm labor and pregnancy outcome, patient inclusion/exclusion characteristics, patient demographic characteristics, drug and cointerventions, and numerous birth, maternal, and neonatal outcome measures. We graded the quality of the individual articles and the strength of the evidence for antibiotic benefit. RESULTS: We abstracted data from 14 randomized trials and 1 observational study. Of these studies, 13 trials met the requirements for a meta-analysis. The meta-analysis demonstrated a mixed outcome pattern with small improvements in pregnancy prolongation, estimated gestational age at birth, and birth weight. Data were insufficient to show a beneficial effect on neonatal morbidity or mortality rates. CONCLUSION: Treatment of preterm labor with antibiotic therapy can prolong gestation. The benefits of antibiotics are small, and there is considerable uncertainty about the optimal agent, route, dosage, and duration of therapy. PMID- 11904630 TI - No headaches! PMID- 11904631 TI - Is male sex an independent risk factor for preterm birth? PMID- 11904632 TI - Poor design in cerclage studies. PMID- 11904633 TI - Multiembryonic pregnancy reduction: the Hungarian experience. PMID- 11904634 TI - The improvement in pregnancy rates with platelet-activating factor in intrauterine insemination was not demonstrated. PMID- 11904635 TI - President's message: the 2002-2005 strategic plan: a foundation for educating the next generation of NAPNAP leaders. PMID- 11904636 TI - Immunization. PMID- 11904637 TI - Pharmaceutical company advertising practices: call to arms. PMID- 11904638 TI - Health care issues for families traveling internationally with children. AB - For a variety of reasons, international travel by American families and their children is increasingly more common. Comfort and health care issues are important to these families, and they often address their questions and concerns to their health care practitioner. Traveling to foreign countries involves concerns about food, water, medications, immunizations, and supplies. Seeking medical care on both a routine and emergency basis may be challenging for families traveling to countries outside the United States. This article discusses health care topics relating to children traveling outside the United States and includes answers to the most commonly asked questions and a list of references and resources for parents and practitioners. Pediatric care providers will find this article to be a helpful guide for their traveling pediatric patients. PMID- 11904639 TI - Educating NPs to educate patients: cholesterol screening in pediatric primary care. AB - INTRODUCTION: This study described practices, knowledge, and attitudes of primary care nurse practitioners (NPs) in Minnesota regarding cholesterol screening in children and adolescents. METHODS: A survey including 22 questions pertaining to cholesterol screening, adapted from a telephone survey used by Arneson, Luepker, Pirie, and Sinaiko (1992), was mailed to pediatric and family NPs. Eighty-three of 221 surveys (38%) were completed, returned, and used for data analysis. RESULTS: Although 96% of the respondents value childhood cholesterol levels as indicators of the risk of developing adult cardiovascular disease, only 64% follow the current recommendation to selectively screen cholesterol levels in patients who have a parent with hypercholesterolemia. Furthermore, only 55% of the respondents screen patients with a family history of premature cardiovascular disease, and only 58% screen patients with other cardiac risk factors. Whereas 57% of respondents correctly identified an acceptable total cholesterol level for children, only 34% correctly identified an acceptable LDL cholesterol level. DISCUSSION: Gaps in knowledge and practice may prevent NPs from implementing recommended guidelines for childhood cholesterol screening. Educating NPs about cholesterol screening is necessary to ensure the comprehensive cardiac health assessment and management of pediatric patients. PMID- 11904641 TI - Parent caregiver-related predictors of health care service utilization by children with cerebral palsy enrolled in Medicaid. AB - INTRODUCTION: This study examined the associations between characteristics of the parent caregiver and health care service utilization by children with cerebral palsy enrolled in Medicaid. METHODS: A questionnaire was administered to the parents of children with cerebral palsy being treated in a North Carolina hospital, and the responses were linked with health services claims data from Medicaid. Data were available for 93 patients who maintained 1 year of continuous enrollment. The impact of selected demographic variables (race, years the child had the condition, employment, transportation) and behavioral variables (belief the child was receiving appropriate care, willingness to use home nursing and respite services, self-assessment of the severity of the child's condition, and use of orthopedic aids) on the frequency and costs of health care services was explored with use of multiple regression analysis. RESULTS: Parents who were willing to use home nursing and respite services were more likely to utilize these services, as well as inpatient facilities and orthopedic care, for their children (P <.05). However, parents who perceived that their child was receiving adequate care were less likely to utilize orthopedic care and home nursing for their children (P <.05). DISCUSSION: The belief of the parents in the adequacy of care received by their child, as well as their willingness to utilize supplemental health care services, were major predictors of health care service utilization by children with cerebral palsy enrolled in Medicaid. Parents who believed that their children receive enough care may need to be targeted for care management and disease management programs to ensure the continuum of treatment and care for these children. PMID- 11904640 TI - Peak flow meters in childhood asthma: parent report of use and perceived usefulness. AB - INTRODUCTION: Peak flow meters (PFMs) in children with moderate to severe asthma have been used to monitor changes in asthma status and inform treatment decisions. However, their usefulness and the likelihood of their long-term use by families remains controversial. METHODS: One hundred sixty-eight children ages 6 to 19 years were enrolled in a longitudinal randomized clinical trial to evaluate the impact of 3 different intensities of symptom monitoring on diverse clinical outcomes: subjective symptom monitoring, symptom-time PFM monitoring, and daily PFM monitoring. RESULTS: At 3 months after the intervention, 90% of parents and 82% of children surveyed perceived a benefit to the monitoring method taught, regardless of group assignment. Ninety-three percent of parents but only 71% of children planned to continue that method. At 1 year after exiting from the study, 69% continued to use a PFM; 30% had discontinued use. No group differences existed in frequency of PFM use between symptom-time and daily users (x = 4.36 vs x = 4.31 times per month). Predictors of continued PFM use included greater frequency of symptoms and younger age. Those discontinuing use believed that it added no additional information to assist in management, using it was a chore/burden, it was not available when needed, and the child's asthma had improved. DISCUSSION: Families will probably use a PFM to inform management during symptomatic times. Daily use is not perceived as useful by most families and is likely to be an unrealistic expectation for most children. J Pediatr Health Care. PMID- 11904642 TI - Increasing antibiotic resistance: its effect on the therapy for otitis media. PMID- 11904643 TI - A 21-month-old girl with hematochezia. PMID- 11904644 TI - Public policy advocacy-why get involved? PMID- 11904645 TI - Helping toddlers eat well. PMID- 11904646 TI - Circle of caring. PMID- 11904648 TI - Equipoise, power, and the pulmonary artery catheter. PMID- 11904649 TI - Saving face: better interfaces for noninvasive ventilation. PMID- 11904650 TI - Evidence-based medicine or fuzzy logic: what is best for ARDS management? PMID- 11904651 TI - C-reactive protein: a valuable marker of sepsis. PMID- 11904652 TI - Evidence-based medicine in the therapy of the acute respiratory distress syndrome. PMID- 11904653 TI - A randomised, controlled trial of the pulmonary artery catheter in critically ill patients. AB - OBJECTIVE: To compare the survival and clinical outcomes of critically ill patients treated with the use of a pulmonary artery catheter (PAC) to those treated without the use of a PAC. DESIGN: Prospective, randomised, controlled, clinical trial from October 1997 to February 1999. SETTING: Adult intensive care unit at a large teaching hospital. PATIENTS: Two hundred one critically ill patients were randomised either to a PAC group ( n=95) or the control group ( n=106). One patient in the control group was withdrawn from the study and five patients in the PAC group did not receive a PAC. All participants were available for follow-up. INTERVENTIONS: Participants were assigned to be managed either with the use of a PAC (PAC group) or without the use of a PAC (control group). MAIN OUTCOME MEASURES: Survival to 28 days, intensive care and hospital length of stay and organ dysfunction were compared on an intention-to-treat basis and also on a subgroup basis for those participants who successfully received a PAC. RESULTS There was no significant difference in mortality between the PAC group [46/95 (47.9%)] and the control group [50/106 (47.6)] (95% confidence intervals for the difference -13 to 14%, p>0.99). The mortality for participants who had management decisions based on information derived from a PAC was 41/91 (45%, 95% confidence intervals -11 to 16%, p=0.77). The PAC group had significantly more fluids in the first 24 h (4953 (3140, 7000) versus 4292 (2535, 6049) ml) and an increased incidence of renal failure (35 versus 20% of patients at day 3 post randomisation p<0.05) and thrombocytopenia ( p<0.03). CONCLUSIONS: These results suggest that the PAC is not associated with an increased mortality. PMID- 11904654 TI - Dobutamine and gastric-to-arterial carbon dioxide gap in severe sepsis without shock. AB - OBJECTIVES: To evaluate the effect of an early dobutamine infusion on gastrointestinal perfusion in patients with severe sepsis. DESIGN: Prospective, randomized, controlled, multicenter clinical study. SETTING: Six medical and/or surgical intensive care units (ICU) of teaching hospitals. PATIENTS: Forty-two patients with severe sepsis. INTERVENTIONS: Patients were divided into two groups according to gastric-to-arterial CO2 gap (DeltaCO2) [normal DeltaCO2 group ( n=17): DeltaCO2 < or = 8 mmHg; increased DeltaCO2 group ( n=25): DeltaCO2 > 8 mmHg]. Patients within each group were then randomized to receive either dobutamine (5 microg/kg per min) or saline for 72 h. MEASUREMENTS AND MAIN RESULTS: SAPS II was similar in both groups [group 1: 44.0 (33.0-56.5); group 2: 48.5 (40.5-59.0), p=0.27]. At ICU admission, mean arterial pressure was lower in the high DeltaCO2 group [73.0 (67.0-79.5) mmHg, p=0.03] than in the normal DeltaCO2 group [84.0 (73.7-104.0) mmHg] while blood lactate [normal DeltaCO2 group: 1.6 (0.8-2.3); high DeltaCO2 group: 1.6 (1.1-1.9) mmol/l] was similar for the two groups. DeltaCO2 was significantly lower in the normal DeltaCO2 group [5.0 (2.0-6.0) mmHg)] than in the high DeltaCO2 group [11.0 (10.0-19.0) mmHg]. Dobutamine infusion did not significantly change hemodynamics, blood lactate concentration or tonometric parameters in any group within the first 72 h and had no particular beneficial effect in this population. CONCLUSIONS: An early infusion of dobutamine at a fixed dose of 5 microg/kg per min during the first 72 h of severe sepsis does not influence gastric DeltaCO2. PMID- 11904655 TI - Combination of venoarterial PCO2 difference with arteriovenous O2 content difference to detect anaerobic metabolism in patients. AB - OBJECTIVE: Under conditions of tissue hypoxia total CO2 production (VCO2) should be less reduced than O2 consumption (VO2) since an anaerobic CO2 production should occur. Thus the VCO(2)/VO(2) ratio, and hence the venoarterial CO2 tension difference/arteriovenous O2 content difference ratio (DeltaPCO2/C(a-v)O2), should increase. We tested the value of the DeltaPCO2/C(a-v)O2 ratio in detecting the presence of global anaerobic metabolism as defined by an increase in arterial lactate level above 2 mmol/l (Lac+). DESIGN AND SETTING: Retrospective study over a 17-month period in medical intensive care unit of a university hospital. PATIENTS: We obtained 148 sets of measurements in 89 critically ill patients monitored by a pulmonary artery catheter. RESULTS: The DeltaPCO2/C(a-v)O2 ratio was higher in those with increased ( n=73) than in the normolactatemic group (2.0+/-0.9 vs. 1.1+/-0.6, p<0.0001). Among all the O2- and CO2-derived parameters the DeltaPCO2/C(a-v)O2 ratio had the highest correlation with the arterial lactate level ( r=0.57). Moreover, for a threshold value of 1.4 the DeltaPCO2/C(a v)O2 ratio predicted significantly better than the other parameters (receiver operating characteristic curves) the presence of hyperlactatemia (positive and negative predictive values of 86% and 80%, respectively). The overall survival estimate at 1 month was greater when the DeltaPCO2/C(a-v)O2 ratio was less than 1.4 on the first set of measurements (38+/-10% vs. 20+/-8%, p<0.01). CONCLUSION: The DeltaPCO2/C(a-v)O2 ratio seems a reliable marker of global anaerobic metabolism. Its calculation would be helpful for a better interpretation of pulmonary artery catheter data. PMID- 11904656 TI - Evaluation of patient skin breakdown and comfort with a new face mask for non invasive ventilation: a multi-center study. AB - OBJECTIVE: To evaluate patient comfort, skin breakdown and eye irritation when comparing a prototype face mask (PM) and conventional face masks (CMs) during non invasive ventilation. SETTING AND DESIGN: Eight centers (intensive or intermediate care units). Multicenter randomized study. POPULATIONS: Patients with acute respiratory failure of different etiologies. INTERVENTIONS: Patients were randomized to CMs or PM when ventilation was expected to last at least 12 consecutive hours a day for two consecutive days. Patient comfort, skin breakdown and eye irritation, assessed by means of standardized scoring systems, were measured after 24 and 48 h and before discontinuing ventilation. RESULTS: Hundred ninety-four patients were randomized. Forty-seven patients were finally enrolled: PM (24) and CMs (23). Ventilator settings were similar in the two groups at the beginning of the treatment and after 24 and 48 h. Skin breakdown was significantly higher in the CMs group over the study period ( p<0.001). Patient comfort was higher in the PM group after 24 and 48 h ( p=0.008 and p<0.001, respectively). Eye irritation was absent in both groups after 24 h and did not differ significantly after 48 h (p=0.539). Before ventilation was discontinued skin breakdown and patient comfort were significantly higher in the CMs group, when compared to the PM group ( p<0.001 and p=0.003, respectively). Eye irritation was slightly higher in the PM versus CMs group ( p=0.21). The time on ventilation was not significantly different between the two groups ( p=0.830). CONCLUSION: The PM significantly reduced skin breakdown while improving patient comfort, compared to the CMs. PMID- 11904657 TI - Suppression of interleukin-6 to interleukin-10 ratio in chronic alcoholics: association with postoperative infections. AB - OBJECTIVE: To investigate the interleukin-6 (IL-6) to interleukin-10 (IL-10) ratio and levels of sE-selectin in patients undergoing elective surgery of the upper digestive tract and to define the differences in the perioperative immune response between chronic alcoholic and non-alcoholic patients. DESIGN: Prospective pilot study. SETTING: Single center, interdisciplinary intensive care unit (ICU) at a university hospital. MEASUREMENT AND MAIN RESULTS: The study compared chronic alcoholics ( n=25) and non-alcoholics ( n=20) before and after surgery for resection of upper digestive tract tumors. White blood cell counts, C reactive protein and circulating levels of sE-selectin, the pro-inflammatory cytokine IL-6 and the inhibitory cytokine IL-10, were obtained at hospital admission, preoperatively, postoperatively at ICU admission and 2 and 4 days later. Rates of postoperative infectious complications including pneumonia and sepsis were determined. sE-selectin only differed between chronic alcoholics and non-alcoholics preoperatively. Compared to non-alcoholics, chronic alcoholic patients showed a fourfold increase in circulating levels of IL-10 ( p<0.01) and a suppression of the IL-6/IL-10 ratio ( p=0.001) immediately after surgery. Coincident with the immune alterations, chronic alcoholics had a prolonged ICU stay ( p<0.01) and a threefold increased rate of wound infections ( p<0.05) and pneumonia ( p<0.01). Lower IL-6/IL-10 ratios were associated with increased rates of infectious complications ( p<0.05). CONCLUSION: Chronic alcoholics had decreased IL-6/IL-10 ratios at ICU admission and increased rates of infectious complications in the postoperative ICU course. This may indicate immediate postoperative immune suppression before the onset of infections and may help to identify chronic alcoholic patients at risk. PMID- 11904658 TI - Amino acid imbalance early in septic encephalopathy. AB - OBJECTIVE: To evaluate plasma amino acid concentrations and markers of inflammation in the early stage and the course of septic encephalopathy. DESIGN: Prospective, case series of patients with well-defined septic encephalopathy. SETTING: Surgical department and intensive care unit of a university hospital. PATIENTS: Seventeen patients with sepsis according to the ACCP/SCCM consensus conference criteria and encephalopathy based on neuropsychological tests, compared to a control group undergoing uncomplicated thoracic surgery. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: SOFA score, blood samples for plasma amino acids, procalcitonin and interleukin-6. Sepsis was determined to be the cause of encephalopathy in 14 of the 17 patients. Six patients developed septic shock, four died within the study period of 28 days. Within 12 h of the onset of septic encephalopathy, mean values of PCT and IL-6 were elevated ( p<0.001) and the amino acids unbalanced (the ratio of branched-chain to aromatic amino acids was decreased, p<0.001). During the course of sepsis the decreased amino acid ratio was significantly, but moderately, correlated with elevated PCT and IL-6 levels. On study days when PCT was higher than 2 ng/ml, the amino acid ratio was significantly lower. In no patient was severe liver dysfunction seen. CONCLUSIONS: Metabolic disturbances with changes in amino acid levels can occur early in septic patients, without serious liver abnormalities. The present data suggest a possible role of amino acids in the pathogenesis of septic encephalopathy. PMID- 11904659 TI - A new simple method for percutaneous tracheostomy: controlled rotating dilation. A preliminary report. AB - OBJECTIVE: To describe and introduce a new technique for percutaneous dilational tracheostomy. DESIGN AND SETTING: Open, observational clinical trial in patients requiring an elective tracheostomy in two intensive care units of university hospitals. PATIENTS: Fifty (25/25) consecutive patients requiring an elective tracheostomy above 18 years of age. INTERVENTIONS: Performance of a percutaneous dilational tracheostomy with a specially designed screw-type dilator, using a thread for the dilation procedure. RESULTS: In 50 consecutive patients the new device allowed a quick and safe dilation procedure without any serious bleeding complications or other relevant procedural-related side effects. CONCLUSIONS: The described new percutaneous dilational tracheostomy device (PercuTwist, Rusch, Kernen, Germany) represents a single-step method with a high degree of control during dilation. So far, it appears to be a safe, quickly performed procedure with a strikingly low incidence of even small bleeding complications, thus offering an interesting new alternative for the performance of a percutaneous tracheostomy. PMID- 11904660 TI - Clinical estimation of trunk position among mechanically ventilated patients. AB - OBJECTIVES: Trunk position at 45 degrees from the horizontal is associated with a decreased risk of gastroesophageal aspiration. The objectives of this study were to determine the accuracy of trunk flexion estimates compared to a reference standard measurement, and to determine agreement about trunk flexion among ICU clinicians. DESIGN: Prospective observational study. SETTING: Two university affiliated medical-surgical ICUs. PATIENTS AND PARTICIPANTS: Thirty-three mechanically ventilated ICU patients, seven residents, two fellows, three intensivists, and twenty-eight bedside nurses. INTERVENTIONS: Prospectively, concurrently, and independently during rounds, one bedside nurse, one resident, one fellow, and one intensivist clinically estimated the trunk flexion of mechanically ventilated patients. To record the reference standard, a trained investigator measured trunk position in the vertical plane using a goniometer. MEASUREMENTS AND RESULTS: We made 438 clinical assessments on 33 patients aged 57.2+/-19.4 (SD) years with an APACHE II score of 27.3+/-9.4. Mean trunk flexion estimates were: nurses 24.3+/-12.3 degrees from the horizontal, residents 20.2+/ 13.7, fellows 20.3+/-10.8, and intensivists 21.1+/-13.1 compared to the reference standard measurement 16.2+/-9.0 degrees. The accuracy of trunk flexion estimates was fair to moderate [intraclass correlation for reference standard versus nurses (ICC 0.42), residents (ICC 0.52), fellows (ICC 0.36), and intensivists (ICC 0.55)]. The agreement among different groups of clinicians was moderate. CONCLUSIONS: In mechanically ventilated patients, we found that clinical estimates of trunk position were moderately good, agreement amongst caregivers was moderately good, but that all clinicians tended to overestimate the angle of semirecumbency. PMID- 11904661 TI - Hypoproteinemia as a marker of acute respiratory distress syndrome in critically ill patients with pulmonary edema. AB - OBJECTIVE: To assess the value of serum protein levels for differentiating permeability pulmonary edema in the course of acute respiratory distress syndrome (ARDS) from cardiogenic pulmonary edema (CPE). DESIGN AND SETTING: Observational cohort study in intensive care units of 720-bed university hospital. PATIENTS: Twenty-four consecutive patients with clinical evidence of edema, 11 fulfilling the consensus definition of ARDS, 7 having sepsis, 5 with all ARDS consensus criteria and sepsis but a pulmonary capillary wedge pressure above 18 mmHg (mixed), and 8 with CPE. All patients except for one with CPE were mechanically ventilated. MEASUREMENTS AND RESULTS: Radionuclide assessments of pulmonary microvascular protein (transferrin) permeability (pulmonary leak index, PLI) were carried out and serum protein levels determined at admission and for ARDS/mixed patients, at recovery, defined by a decrease in positive end-expiratory pressure to 0 cmH2O. At admission the PLI was higher in ARDS/mixed than in CPE patients. The total protein and transferrin levels were lower in the former. The area under the curve of the receiver operating characteristic for diagnosing ARDS (vs. CPE) was 0.98 for transferrin (cutoff value 1.5 g/l), 0.95 for total protein (cutoff value 59 g/l) and 0.80 for albumin (cutoff value 24 g/l) levels. In various clinical diagnostic groups the transferrin level approached the PLI in diagnostic value. At recovery the PLI had decreased and serum protein levels increased. CONCLUSIONS: The data suggest that hypoproteinemia is a marker of ARDS. This may partially reflect increased permeability in the lungs, systemically, or both. PMID- 11904663 TI - Analysis of resource use and cost-generating factors in a German medical intensive care unit employing the Therapeutic Intervention Scoring System (TISS 28). AB - OBJECTIVE: Evaluation of resource use and costs of a medical intensive care unit (ICU) utilising the simplified Therapeutic Intervention Scoring System (TISS-28). DESIGN: Prospective observational study. SETTING: : Medical ICU of a tertiary care centre. PATIENTS: Consecutive patients with an ICU length of stay (LOS) more than 24 h. INTERVENTIONS: Over a 3 month period SAPS II, TISS-28 and SOFA were determined daily. Patients were retrospectively classified as receiving active (AT) or non-active (NAT) treatment according to TISS-28 variables, with AT representing a therapeutic intervention that could not be performed outside the ICU. Individual expenditure for all patients was calculated using a modified 'top down' method. MEASUREMENTS AND RESULTS: Out of 303 consecutive patients, 241 (79.5%), including all non-survivors, were categorised AT. The hospital mortality was 14.5%. TISS-28 and ICU LOS were higher in patients receiving AT ( p<0.001). Patient-specific costs accounted for 36 EUR per TISS-point and daily treatment costs 1336 EUR for all patients. Daily costs of care were 68 EUR higher for AT, compared to NAT, patients ( p<0.001). There was no association between ICU costs and measures of severity of illness (SAPS II, SOFA). CONCLUSIONS: TISS-28 is a fast, reliable and readily applicable tool to identify patients receiving AT. Although total and daily costs of care were significantly higher in patients receiving AT, the difference of the daily costs was, albeit statistically significant, economically negligible. The main difference in ICU costs was attributable to ICU LOS. Therefore cost-saving strategies must aim at reducing ICU LOS, without compromising quality of care. PMID- 11904664 TI - Effects of price information on test ordering in an intensive care unit. AB - OBJECTIVE: To determine if daily information on the price of common laboratory tests and chest X-ray could significantly influence test ordering by physicians and decrease the costs. DESIGN: A prospective observational and sequential study. SETTING: A 21-bed surgical intensive care unit of a university hospital. PATIENTS: All patients admitted during a 4-month period. INTERVENTIONS: A 2-month period served as control (period I). During a consecutive 2-month period (period II) physicians were informed about the costs of seven common diagnostic tests (plasma and urinary electrolytes, arterial blood gases, blood count, coagulation test, liver function test and chest X-ray). The number of tests ordered and costs during the two periods were compared. MEASUREMENTS AND RESULTS: A total of 287 patients were included (128 in period I and 159 in period II). Information about age, gender, Severe Acute Physiologic Score II, McCabe score, intensive care unit length of stay and mortality were collected and were not statistically different between the two study periods. Except for liver function tests, all the tests evaluated were less frequently prescribed when physicians were aware of the charges, irrespective of whether the tests were routine or requested during an emergency. Nevertheless, a significant reduction was obtained only for arterial blood gases and urinary electrolytes. Overall analysis of the expenditures (in Euros) showed a significant 22% decrease in period II (341+/-500 versus 266+/-372 Euros, p<0.05). CONCLUSION: Providing price information to physicians was associated with a significant reduction for arterial blood gases and urinary electrolytes tests ordered and was significantly cost-saving. PMID- 11904662 TI - Evaluation of a new device for noninvasive measurement of nonshunted pulmonary capillary blood flow in patients with acute lung injury. AB - OBJECTIVES: To evaluate the performance of a new device for noninvasive measurement of nonshunted pulmonary capillary blood flow (PCBF) by partial CO2 rebreathing. DESIGN AND SETTING: Prospective clinical trial in an intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: Twenty mechanically ventilated patients with acute lung injury. INTERVENTIONS: Variations in PEEP of +/-3 cmH2O. MEASUREMENTS AND RESULTS: Initially PCBF was measured invasively as cardiac output minus venous admixture (Q(VA)/Q(t)) flow, and by partial CO2 rebreathing at baseline PEEP (PEEP(b)). The PEEP was then reduced by 3 cmH2O (to PEEP(b-3)) and measurements were repeated after 30 min. PEEP was then increased by 6 cmH2O (to PEEP(b+3)), and measurements were repeated after 10, 20, and 30 min. The overall correlation coefficient between noninvasive and invasive PCBF measurements at PEEP(b) was high ( r=0.97), with close agreement between methods being observed (0.1+/-0.6 l/min, bias and precision, respectively). Accordingly, both the correlation coefficient and agreement between methods for changes in PCBF from PEEP(b-3) to PEEP(b+3) levels were satisfactory ( r=0.71; 0.2+/-0.5 l/min, bias and precision). The new device was able to detect the correct PCBF trend in 17 of 20 patients investigated and in all patients who showed invasive PCBF changes equal to or greater than 0.3 l/min ( n=12). Noninvasive PCBF changes were stable as early as 10 min after variation in PEEP, as compared to 30 min values. CONCLUSIONS: The new device appears to be clinically useful for the monitoring of PCBF in patients suffering from acute lung injury. Our results suggest that titration of PEEP aimed at improving PCBF can be performed with the new device. PMID- 11904665 TI - Amylin is associated with delayed gastric emptying in critically ill children. AB - OBJECTIVES: Amylin is a novel 37 amino acid that is secreted together with insulin from the pancreas in response to enteral nutrient intake. As a potent inhibitor of gastric motility it plays an important role in the control of carbohydrate absorption. In this study we aimed to determine the relationship between amylin levels and gastric emptying in critically ill children. DESIGN: Prospective interventional study. SETTING: Tertiary paediatric intensive care unit. PATIENTS: Twenty-three patients were studied following admission to a paediatric intensive care unit. The median age (25th-75th centiles) was 5.8 years (1.5-11.6) and weight 20 kg (12.8-47.5). INTERVENTIONS: Patients were defined as feed-intolerant on the basis of gastric residual volume greater than 125% 4 h after a feed challenge. Three objective measures of gastric emptying were then calculated from a 6 h paracetamol absorption test. Blood glucose, serum insulin and amylin levels were averaged across the paracetamol absorption test period. MEASUREMENTS AND RESULTS: Eight patients were classified as feed-intolerant (nTOL) and 15 as feed-tolerant (TOL) [median gastric residual volumes 321% (261 495) and 4% (0-6), respectively]. Gastric emptying was delayed in the feed intolerant group as assessed by all paracetamol absorption test parameters ( p< or =0.01). The median serum amylin concentration was significantly higher in the feed-intolerant group [nTOL 47.0 (37.7-54.8) versus TOL 22.7 (13.6-26.7) pmol/l, p<0.0001]. A positive correlation between serum amylin and insulin was observed ( r=0.46, p=0.02) but not between amylin and glucose ( r=0.25, p=0.23). CONCLUSIONS: The use of gastric residual volumes to define feed intolerance is justified in critically ill children. High serum amylin levels are associated with delayed gastric emptying in these patients. The correlation between serum amylin and insulin levels indicates a degree of preservation of pancreatic hormonal co-release. PMID- 11904666 TI - A new prognostic scoring system for meningococcal septic shock in children. Comparison with three other scoring systems. AB - OBJECTIVE: To develop a quick and sensitive method for identification of children with presumed meningococcal septic shock at risk of death at admission to the pediatric intensive care unit (PICU) and to compare its performance with three other prognostic systems: Glasgow Meningococcal Septicaemia Prognostic Score (GMSPS), Malley score and the Paediatric Index of Mortality (PIM). DESIGN: Multicenter retrospective cohort study. SETTING: PICUs of 14 tertiary hospitals. PATIENTS: The developmental sample included 192 children consecutively admitted to the PICUs with presumed or confirmed meningococcal septic shock from 1983 to 1995. The validation sample included 158 children consecutively admitted from 1996 to 1998. INTERVENTIONS: Clinical and laboratory data gathered during the first 2 h after admission were used to develop the new score and to compute the other scoring systems. Logistic regression was applied to identify the independent predictors of death. MEASUREMENTS AND RESULTS: Overall mortality was 31.5%. The new score included seven variables: cyanosis (2 points), Glasgow coma scale less than 8 (2 points), refractory hypotension (2 points), oliguria (1 point), leukocytes less than 4000/mm(3) (1 point), partial thromboplastin time more than 150% of control value (1 point) and base deficit more than 10 mmol/l (1 point). The new score provided the best discriminative capability, as measured by the area under the ROC curve (SEM) in the validation sample =0.88 (0.03), PIM =0.82 (0.04), Malley I =0.80 (0.04), GMSPS =0.79 (0.04) and Malley II =0.76 (0.04). CONCLUSIONS: A new prognostic score is proposed for therapeutic stratification of children with presumed meningococcal septic shock. PMID- 11904667 TI - Inhaled budesonide in experimental chlorine gas lung injury: influence of time interval between injury and treatment. AB - OBJECTIVE: To examine the time window between injury and treatment during which nebulized corticosteroid lessens lung injury induced by chlorine gas inhalation. DESIGN: An experimental laboratory study. SETTING: Academic research laboratory. SUBJECTS: Twenty-four juvenile female pigs. INTERVENTIONS: Twenty-four mechanically ventilated pigs were exposed to chlorine gas (400 PPM in air) for 20 min, then divided into four groups (six in each group). Nebulized budesonide (BUD) was given immediately (BUD 0 min), 30 min (BUD 30 min) or 60 min (BUD 60 min) after chlorine gas exposure. Six pigs receiving nebulized saline served as controls. MEASUREMENTS AND MAIN RESULTS: Hemodynamics, gas exchange and lung mechanics were evaluated for 5 h after chlorine gas exposure. All animals had an immediate increase in airway and pulmonary artery pressure and a sharp drop of arterial oxygenation. The mean arterial oxygen tension of BUD 0 min and BUD 30 min animals was significantly higher than in the control and the BUD 60 min groups ( p<0.001). The recovery of lung compliance in the BUD 0 min and the BUD 30 min groups was significantly more rapid than in the control and the BUD 60 min groups ( p<0.001). The pulmonary wet to dry weight ratio was greater in the control group than in the BUD-treated groups ( p<0.05). CONCLUSION: Treatment with inhaled budesonide immediately or 30 min after chlorine gas lung injury had similar positive effects on symptoms and signs of pulmonary injury, whereas treatment delayed for 60 min was less effective. PMID- 11904669 TI - One-year survival and neurological outcome after pediatric cardiopulmonary resuscitation. AB - OBJECTIVE: Reported survival after cardiopulmonary resuscitation (CPR) in children varies considerably. We aimed to identify predictors of 1-year survival and to assess long-term neurological status after in- or outpatient CPR. DESIGN: Retrospective review of the medical records and prospective follow-up of CPR survivors. SETTING: Tertiary care pediatric university hospital. PATIENTS AND METHODS: During a 30-month period, 89 in- and outpatients received advanced CPR. Survivors of CPR were prospectively followed-up for 1 year. Neurological outcome was assessed by the Pediatric Cerebral Performance Category scale (PCPC). Variables predicting 1-year survival were identified by multivariable logistic regression analysis. INTERVENTIONS: None. RESULTS: Seventy-one of the 89 patients were successfully resuscitated. During subsequent hospitalization do-not resuscitate orders were issued in 25 patients. At 1 year, 48 (54%) were alive, including two of the 25 patients with out-of-hospital CPR. All patients died, who required CPR after trauma or near drowning, when CPR began >10 min after arrest or with CPR duration >60 min. Prolonged CPR (21-60 min) was compatible with survival (five of 19). At 1 year, 77% of the survivors had the same PCPC score as prior to CPR. Predictors of survival were location of resuscitation, CPR during peri- or postoperative care, and duration of resuscitation. A clinical score (0 15 points) based on these three items yielded an area under the ROC of 0.93. CONCLUSIONS: Independent determinants of long-term survival of pediatric resuscitation are location of arrest, underlying cause, and duration of CPR. Long term survivors have little or no change in neurological status. PMID- 11904668 TI - Effects of prolonged mechanical ventilation and inactivity on piglet diaphragm function. AB - OBJECTIVES: Muscle weakness is associated with immobilization, prolonged mechanical ventilation, critical illness and various critical care therapies. This study used an animal model simulating the critical care environment to investigate the effects of 5 days' mechanical ventilation and inactivity on diaphragm contractility and neurophysiologic function. DESIGN: Prospective laboratory study. SETTING: Animal research laboratory. SUBJECTS: Seven 2-3 month old piglets weighing 20-25 kg. INTERVENTIONS: The animals received constant-flow, volume-controlled mechanical ventilation (Tv 12-15 ml/kg, PEEP 3-5 cmH2O, I:E 1:2) and sedation without paralysis, and spontaneous breathing efforts were prevented. Evoked diaphragm contractions were achieved by transvenous phrenic nerve pacing. MEASUREMENTS AND MAIN FINDINGS: Transdiaphragmatic pressure (Pdi) measurements were used to assess force frequency relationships. Evoked electrophysiologic measures included lowest stimulus threshold and latency, compound muscle action potential (CMAP) amplitude and duration, and amplitude during repetitive nerve stimulation at 3 Hz. Lung function measures included airway pressures, tidal and minute volumes, and dynamic compliance and resistance. There were no clinically significant changes in hemodynamics, oxygenation or ventilation. Indirect measures of lung volume remained stable. Pdi decreased by 20% at all frequencies tested and was accompanied by a 30% decrease in evoked CMAP amplitude, (6.7+/-4.7 mV to 4.5+/-3.9 mV, p=0.01) while CMAP threshold, latency and duration were unchanged and no significant decrement in amplitude was seen during repetitive stimulation at 3 Hz. CONCLUSION: In this in vivo model of prolonged mechanical ventilation in an intensive caring setting, 5 days of mechanical ventilation with sedation and complete diaphragm inactivity resulted in disturbed diaphragm contractility and activation, while nerve conduction and neuromuscular transmission were not affected. Based on these findings, it is likely that the changes seen occur at the level of peripheral muscle. PMID- 11904671 TI - Re-expansion pulmonary edema with normal pulmonary artery occlusion pressure during liver transplantation. PMID- 11904672 TI - Dear PCT, you are not a specific marker of bacterial infection. PMID- 11904670 TI - Mechanisms of organ dysfunction in critical illness: report from a Round Table Conference held in Brussels. PMID- 11904673 TI - Massive septic thrombus formation on a superior vena cava indwelling catheter following Torulopsis (Candida) glabrata fungemia. AB - Fungal endocarditis is an exceedingly rare complication of indwelling central venous catheters in adults. Here we describe what appears to be the first case of a right atrial thrombus superinfected with the yeast Torulopsis (Candida) glabrata and attached to an indwelling superior vena cava catheter that was not used for parenteral nutrition. A large vegetation-like mass adherent to the catheter tip was visualized by transesophageal echocardiography in a patient who presented with signs of septic pulmonary embolism. Following open-heart surgery, the definitive diagnosis was established by histopathologic examination of the surgical specimen. PMID- 11904675 TI - Identification of an S-adenosylhomocysteine hydrolase-like transcript induced during dendritic cell differentiation. AB - Dendritic cells (DC) are the professional antigen-presenting cells that initiate immune responses. While DC take up antigen, migrate to lymph nodes and present processed antigen to T lymphocytes, little is known of the intracellular biochemical pathways controlling these events. Using the differential display technique, employing the activated blood DC-like cell line L428, we isolated a cDNA induced during DC differentiation likely to have a regulatory function. This cDNA encoded a putative 530-amino-acid (aa) protein consisting of a unique hydrophilic domain (106 aa) and a domain (424 aa) similar to the methylation pathway enzyme S-adenosylhomocysteine hydrolase (AHCY). Therefore, this molecule was termed DC-expressed AHCY-like molecule (DCAL). DCAL mRNA was expressed moderately in fresh blood DC, but was not detectable in other peripheral blood mononuclear cells. DCAL mRNA increased markedly during activation of blood and skin DC (Langerhans cells), but was diminished in terminally differentiated tonsil DC. Cultured monocytes expressed little DCAL mRNA, but levels increased markedly when differentiated into DC by cytokines GM-CSF and IL-4. The DCAL gene [Chromosome (Chr) 1] and another previously identified DCAL-like molecule KIAA0828 (Chr 7) differed from the AHCY gene (Chr 20) in gene organization. Thus, DCAL may have a role in controlling critical events in DC differentiation and belong to a novel family of AHCY-like molecules. PMID- 11904676 TI - Co-selection of the H63D mutation and the HLA-A29 allele: a new paradigm of linkage disequilibrium? AB - The major histocompatibility complex (MHC) shows a remarkable conservation of particular HLA antigens and haplotypes in linkage disequilibrium in most human populations, suggesting the existence of a convergent evolution. A recent example of such conservation is the association of particular HLA haplotypes with the HFE mutations. With the objective of exploring the significance of that association, the present paper offers an analysis of the linkage disequilibrium between HLA alleles or haplotypes and the HFE mutations in a Portuguese population. Allele and haplotype associations between HLA and HFE mutations were first reviewed in a population of 43 hemochromatosis families. The results confirmed the linkage disequilibrium of the HLA haplotype HLA-A3-B7 and the HLA-A29 allele, respectively, with the HFE mutations C282Y and H63D. In order to extend the study of the linkage disequilibrium between H63D and the HLA-A29-containing haplotypes in a normal, random population, an additional sample of 398 haplotypes was analyzed. The results reveal significant linkage disequilibrium between the H63D mutation and all HLA-A29-containing haplotypes, favoring the hypothesis of a co selection of H63D and the HLA-A29 allele itself. An insight into the biological significance of this association is given by the finding of significantly higher CD8(+) T-lymphocyte counts in subjects simultaneously carrying the H63D mutation and the HLA-A29 allele. PMID- 11904678 TI - Polymorphisms of the TNF gene and the TNF receptor superfamily member 1B gene are associated with susceptibility to ulcerative colitis and Crohn's disease, respectively. AB - The importance of tumor necrosis factor (TNF)-alpha and the TNF receptor gene polymorphisms in the etipathogenesis of inflammatory bowel disease (IBD) has not been elucidated. DNA from peripheral blood samples was obtained from 124 patients with Crohn's disease (CD), 106 patients with ulcerative colitis (UC), and 111 unrelated healthy controls. We examined two single nucleotide polymorphisms (SNPs) of the TNF-alpha gene, TNF (-308 G/A and -238 G/A), an SNP of the TNF receptor superfamily member 1A gene, TNFRSF1A(also known as TNFR1), at codon 12 in exon 1 (CCA/CCG), and two SNPs of the 1B gene, TNFRSF1B (also known as TNFR2), (1466 A/G and 1493 C/T). There was a difference in the carrier frequency for haplotype AG (-308 A, -238 G) between UC patients and the controls (OR=4.76, 95% CI=1.53-14.74, P<0.01). We found a significant difference in carrier frequency for haplotype AT (1466 A, 1493 T) of the TNFRSF1B gene between CD patients and the controls (OR=2.13, 95% CI=1.08-4.21, P<0.05). The significance proved to be greater in CD patients with both internal and external fistula (OR=4.8, 95% CI=1.73-13.33, P<0.01), and in those who were poor responders ( n=22) to our treatments, which consisted of nutritional therapy, medical therapy and surgical therapy (OR=9.24, 95% CI=3.37-25.36, P<0.001). This study suggests that one of the genes responsible for UC may be the TNF gene, or an adjacent gene, and that TNFRSF1B gene polymorphisms contribute greatly to the increased onset risk of CD and to the disease behavior. PMID- 11904677 TI - Distinctive KIR and HLA diversity in a panel of north Indian Hindus. AB - HLA and KIR are diverse and rapidly evolving gene complexes that work together in human immunity mediated by cytolytic lymphocytes. Understanding their complex immunogenetic interaction requires study of both HLA and KIR diversity in the same human population. Here a panel of 72 unrelated north Indian Hindus was analyzed. HLA- A, B, C, DRB1, DQA1, and DQB1 alleles and their frequencies were determined by sequencing or high-resolution typing of genomic DNA; KIR genotypes were determined by gene-specific typing and by allele-level DNA typing for KIR2DL1, 2DL3, 2DL5, 3DL1, and 3DL2. From HLA analysis, the north Indian population is seen to have several characteristics shared either with Caucasian or East Asian populations, consistent with the demographic history of north India, as well as specific features, including several alleles at high frequency that are rare or absent in other populations. A majority of the north Indian KIR gene profiles have not been seen in Caucasian and Asian populations. Most striking is a higher frequency of the B group of KIR haplotypes, resulting in equal frequencies for A and B group haplotypes in north Indians. All 72 members of the north Indian panel have different HLA genotype and different KIR genotype. PMID- 11904679 TI - Genetic polymorphism of the human ICOS gene. AB - Inducible costimulator (ICOS) is a novel receptor belonging to the same family as CD28 and CTLA4, which regulate T-lymphocyte activation in the immune response. The genes for these molecules are located adjacent to each other on Chromosome 2q33. Many autoimmune diseases have been found to be genetically linked to or associated with genetic markers near the CTLA4 gene. However, as all three genes are closely linked and have related functions, it is possible that the findings could be explained by variation in CD28 or ICOS. Few data on genetic variation in the ICOS gene are available. We sequenced the ICOS gene in 13 healthy unrelated individuals and found eight single nucleotide polymorphisms. One was located in the first intron, and the others in the untranslated region of the last exon. The allele frequencies and linkage disequilibrium were determined from a population sample of 63 Finnish individuals. The results show that the ICOS gene is polymorphic, but no variation in the coding sequence was detected, implying that the genetic linkage of this gene region to autoimmune diseases may not result from structural variation in the ICOS molecule. These polymorphisms, however, should be useful in genetic studies of this candidate gene. PMID- 11904680 TI - Mouse IgA allotypes have major differences in their hinge regions. AB - Six IgA allotypes are serologically identifiable in inbred mice. The sequences of the PCR-amplified C alpha 1, C alpha 2 and C alpha 3 exons from the genomic DNA of mice of four previously unsequenced allotypes already have been compared with those of BALB/c and of a wild mouse, Mus pahari, in the literature. Sporadic differences, including several that may encode the known allotypic determinants, are found throughout the three exons, but major differences occur in the hinge. The hinge is longest in DBA/2 ( Igh-2(c)) mice, having an extra codon compared with that of BALB/c ( Igh-2(a)) and B10.A ( Igh-2(b)) mice. It is two codons shorter in CE ( Igh-2(f)) and four shorter in M. pahari, AKR and NZB (both Igh 2(d)) mice, but the position of the missing codons in the latter two strains is offset from that in M. pahari. The hinges in BALB/c ( Igh-2(a)) and DBA/2 ( Igh 2(c)) differ most from each other and from the other three allotypes, which are fairly closely related. Both BALB/c and DBA/2 have O-linked glycosylation sites, but they are in different positions in the hinge. Compared with BALB/c ( Igh 2(a)), B10.A( Igh-2(b)) has two extra Cys residues in the hinge, while DBA/2 ( Igh-2(c)), AKR/NZB ( Igh-2(d)) and CE ( Igh-2(f)) each have one. The differences in hinge length may have arisen by mismatching of highly repetitive portions of its sequence during meiotic recombination. Possible effects of the differences in hinge length and composition on the behavior of the mouse IgA allotypes are discussed. PMID- 11904681 TI - Does the rat have an H2-D orthologue next to Bat1? AB - Unlike all other mammalian species, which have only one class I region, rat and mouse possess a second class I region on the centromeric side of the MHC. The mouse has class Ia genes in both the centromeric H2-K and the telomeric H2-D region, whereas the rat has class Ia genes only in the centromeric RT1.A region. Bat1 is the last gene of the class III region in the mouse, and H2-D was mapped 10 kb telomeric of Bat1. To determine whether the rat possesses an H2-D orthologue, we sequenced a cosmid clone that contains rat Bat1 and an adjacent class I gene, RT1.46 (l). Homology searches suggest a transition in the rat sequence with a proximal stretch containing Nfkbil1, ATP6G, and Bat1, which is homologous to the mouse H2-D region, and a more-distal stretch, which contains the class I gene and has many similarities to mouse H2-Q region sequences. Downstream of Bat1 is a sequence stretch with great similarity to intron 3 of H2 D, which is not present in any of the rat class I genes but is found in mouse H2 K, D-, and - Q region genes. Numerous repetitive elements indicate that the region is prone to repeat-mediated rearrangements. A putative H2-D orthologue may have been present at this location and lost by genomic rearrangements, leaving the short intronic sequence behind. The class I gene RT1.46 (l) has an open reading frame, but it is unlike H2-D due to a unique 5'UTR shared with H2-Q1 and Q2, the absence of the B2 SINE repeat characteristic of H2-D/L, and the apparent lack of surface expression. We conclude that at least the LEW rat (RT1 (l)) does not possess an H2-D orthologue. PMID- 11904682 TI - Polymorphism and evolution in the constant region of the T-cell receptor beta chain in an advanced teleost fish. AB - Sequence, PCR and Southern data are presented as evidence that, as in mammals, two gene loci encode C regions of the TCR beta chain in the bicolor damselfish, Stegastes partitus. The loci are distinguished by an insertion of ten amino acids in the c-d loop at one locus and by a high interlocus divergence of the third intron and fourth exon sequences. Unlike their mammalian counterparts, the damselfish TCRBC genes encode highly polymorphic regions. None of the eight complete cDNA or four partial genomic DNA sequences presented from a single animal are identical; and three of the four animals examined are heterozygous at both loci, suggesting high heterozygosity in the damselfish population. Coding regions of the eight cDNA clones differ by up to 12% at the DNA level and 23% at the amino acid level. Polymorphism is concentrated primarily in the less evolutionarily conserved regions, suggesting that this variation may be selectively neutral. However, a comparison of the variation between synonymous and non-synonymous sites suggests that at least a portion of the observed variation results from selection. As in mammals, a gradient of sequence homogenization between the two loci is observed. Data presented here suggest that both interlocus homogenization and the sharing of polymorphic segments are likely achieved by partial gene conversion. PMID- 11904684 TI - Characterization of immunoglobulin gamma 1 from a monotreme, Tachyglossus aculeatus. AB - A full-length immunoglobulin gamma clone from the echidna (Tachyglossus aculeatus) was isolated from a spleen cDNA library. The clone was 1,664 base pairs long and encoded the entire open reading frame, incorporating the V, D, J and C regions. The echidna clone had approximately 41% identity and 67% similarity at the amino acid level with both marsupial and eutherian IgG molecules. The presence of IgG in the monotremes confirms that the appearance of IgG occurred prior to the separation of the three extant mammalian lineages, but after their separation from the reptilian lineage, pinpointing the date to between 310 and 170 million years ago. Phylogenetic analyses using the immunoglobulin sequence data strongly support the 'Theria' hypothesis, with the monotreme lineage diverging prior to the separation of the marsupial and eutherian lineages. PMID- 11904683 TI - Complement in urochordates: cloning and characterization of two C3-like genes in the ascidian Ciona intestinalis. AB - The recent identification of complement components in deuterostome invertebrates has indicated the presence of a complement system operating via an alternative pathway in echinoderms and tunicates and via a MBL-mediated pathway thus far identified only in tunicates. Here, we report the isolation of two C3-like genes, CiC3-1 and CiC3-2, from blood cell total RNA of the ascidian Ciona intestinalis. The deduced amino acid sequences of both Ciona C3-like proteins exhibit a canonical processing site for alpha and beta chains, a thioester site with an associated catalytic histidine and a convertase cleavage site, thus showing an overall similarity to the other C3 molecules already characterized. Southern blotting analysis indicated that each gene is present as a single copy per haploid genome. In situ hybridization experiments showed that both CiC3-1 and CiC3-2 are expressed in one type of blood cell, the compartment cells. Two polyclonal antibodies, raised against two deduced peptide sequences in the alpha chain of CiC3-1 and CiC3-2, allowed the identification by Western blot of a single band in the blood serum, of about M(r)150,000. A phylogenetic tree, based on the alignment of CiC3-1 and CiC3-2 with molecules of the alpha(2) macroglobulin superfamily, indicated that the Ciona C3s form a cluster with Halocynthia roretzi C3. The phylogenetic analysis also suggested that the duplication event from which the CiC3-1 and CiC3-2 genes originated occurred in the urochordate lineage after the separation of the Halocynthia and Ciona ancestor. PMID- 11904686 TI - The accuracy of imaging in the local staging of appendicular osteosarcoma. AB - Appendicular osteosarcoma represents the commonest malignant bone tumour for which limb salvage surgery and endoprosthetic replacement is performed. These techniques have been made possible due to the introduction of effective neoadjuvant chemotherapy and demand a high degree of accuracy in the assessment of local staging. This is now achieved with a combination of imaging techniques, the most important of which is magnetic resonance imaging (MRI). For the purposes of local staging, it is useful to divide disease extent into intramedullary and extramedullary. Intramedullary extension includes an assessment of longitudinal medullary extent, epiphyseal involvement and skip metastases. These factors will determine the site of bone resection. Extramedullary extension includes an assessment of joint involvement, relationship to the neurovascular bundle and involvement of muscle compartments. These factors will determine the feasibility and type of limb salvage performed. This article reviews the accuracy of imaging in the local staging of appendicular osteosarcoma. PMID- 11904687 TI - Transient bone marrow edema of the talus: MR imaging findings in five patients. AB - OBJECTIVE: To describe the MR findings of transient bone marrow edema (TBME) of the talus and to address the differential diagnostic considerations. DESIGN AND PATIENTS: The imaging findings of TBME of six tali were retrospectively reviewed in five patients with a clinical history of pain without trauma. Inclusion criteria were MR imaging findings that, when compared with clinical data and results of follow-up assessment, allowed the diagnosis of TBME. MR imaging, standard radiography, and bone scintigraphy were performed. The images were reviewed with particular attention to the pattern and distribution of abnormal marrow signal intensity as well as associated findings. RESULTS: In four cases the entire talus was involved, and in two cases only a portion of the bone was affected. No fractures were detected. MR imaging demonstrated diffuse decreased signal intensity of the marrow on T1-weighted images with corresponding increased signal intensity on T2-weighted images. In all six cases MR imaging detected associated findings, which included joint effusion and soft tissue edema. All patients improved clinically with conservative therapy over a period of 6 months to 1 year. CONCLUSIONS: Although unusual, TBME can involve the talus. Marrow edema without evidence of a fracture and in the absence of history of trauma is a characteristic MR imaging feature, allowing confident diagnosis and institution of conservative therapy. PMID- 11904688 TI - The utility of MR in assessing Blount disease. AB - OBJECTIVE: To review the MR appearances of Blount disease. DESIGN AND PATIENTS: The MR examinations of six knees in four patients (ages 6-7 years) with Blount disease were reviewed. RESULTS: All patients showed delay in ossification of the medial tibial epiphysis. A spectrum of changes was seen in and around the tibial growth plate including: widening and depression of the medial growth plate; small and deep intrusions of cartilage into the metaphysis; edema of the medial tibial epiphysis and medial and lateral metaphysis; varus deformity of the lower leg; widening of the lateral growth plate; osteochondral injury to the medial femoral condyle; hypertrophy of the medial meniscus; focal bone bridging. CONCLUSION: MR appearances are consistent with the primary abnormality in Blount disease, which is failure of endochondral ossification of the medial growth plate. MR examination is useful in surgical planning. PMID- 11904689 TI - Oblique radiograph for the detection of bone spurs in anterior ankle impingement. AB - OBJECTIVE: The aim of this study was to develop a radiographic view to detect anteromedial talotibial osteophytes that remain undetected on standard radiographs. DESIGN AND PATIENTS: In 10 cadaver specimens the maximal size was measured of anteromedial tibial osteophytes that remain undetected on a standard lateral radiograph projection, due to the presence of the anteromedial tibial rim. The average projection of the most prominent anterolateral tibial rim over the anteromedial rim was found to be 7.3 mm. A 7 mm barium-clay osteophyte was attached to this anteromedial rim of the distal tibia. Anteromedial osteophytes become most prominent on an oblique view, in which the radiographic beam is tilted into a 45 degrees craniocaudal direction with the leg in 30 degrees external rotation. This oblique view was compared with the findings of arthroscopic surgery in 25 consecutive patients with anterior ankle impingement syndrome. RESULTS: Medially located tibial and talar osteophytes remained undetected on a standard lateral projection and became visible on the oblique anteromedial impingement (AMI) radiograph. Anterolateral tibial and talar osteophytes were well detected on a standard lateral radiograph projection but were invisible on the AMI view. There was a high correlation between the location of the osteophyte and the location of symptoms and the findings at arthroscopy. CONCLUSION: A combination of lateral and oblique radiographs can be used to differentiate between anteromedial and anterolateral bony ankle impingement. PMID- 11904690 TI - Kashin-Beck disease in children: radiographic findings in the wrist. AB - OBJECTIVE: To characterize the features and prevalence of radiographic abnormalities of the wrist in children with Kashin-Beck disease (KBD) and to determine whether the presence of radiographic abnormalities in the wrist correlates with the severity of KBD. DESIGN AND PATIENTS: Two hundred and eight posteroanterior radiographs of the right hand (including wrist) in children with KBD, ranging in age from 4 to 11 years (mean age 7.7 years), from endemic areas of China were reviewed. Carpal bony margins were evaluated for blurring, thinning, irregularity with and without sclerosis, interruption, depression or destruction. The radiocarpal, intercarpal and carpometacarpal joints were assessed for widening or narrowing. The severity of the disease was graded using the hand criteria from the Chinese Radiographic Criteria of KBD Diagnosis, which classifies the following five types according to the location of the hand involved: I, metaphysis; II, diaphysis; III, I+II; IV, metaphysis and epiphysis; V, II+IV. RESULTS: Of the 208 children, 95 had abnormalities in the hand but not in the wrist; 108 had both hand and wrist abnormalities; only five had abnormal wrist findings without any hand abnormalities. Of the 108 cases with wrist abnormalities, all the carpal bones were involved in 33 cases, of which the hand types were either IV or V. However, any individual carpal bone, or combination of bones, may become involved. The carpal bones most likely to show abnormalities were the capitate and the hamate (93%), followed by the triquetrum (31%), the lunate (9%), the scaphoid (6%), and the trapezoid and the trapezium (5%). The pisiform bones were not evaluated because they cannot be seen on the overlapping posteroanterior radiographs. The most commonly involved carpal joint was the midcarpal joint (42%). CONCLUSIONS: Recognizing carpal abnormalities on radiographs is helpful for the diagnosis of KBD and the evaluation of the severity of the disease. The more severe the KBD, the more likely that the carpal bones will be involved. The capitate and hamate are frequently affected if the disease involves the carpal bones. PMID- 11904691 TI - Granulomatous myositis: a manifestation of chronic graft-versus-host disease. AB - A case of granulomatous myositis as a manifestation of chronic graft-versus-host disease is presented. The clinical presentation, MR imaging appearances, pathologic features and excellent response to treatment with immunosuppression are described. To the best of our knowledge, based on a world literature search, this is the first report of graft-versus-host disease presenting as granulomatous polymyositis., PMID- 11904692 TI - Both Trevor and Ollier disease limited to one upper extremity. AB - A case is presented of a young boy in whom features of Trevor's dysplasia epiphysealis hemimelica and Ollier's enchondromatosis coexisted in a single extremity, the right upper. As Trevor disease consists of osteochondromas of epiphyses and their equivalents, such as carpal and tarsal bones, it is of interest that exostosis-like centers of the neck of radius and perhaps the proximal third metacarpal are present as well. Advanced maturation of selected centers was most marked at the right scaphoid. The child shows alignment abnormalities as a consequence of the varied lesions, including a varus of the right wrist. The coexistence of these varied osteochondromatous abnormalities in one extremity suggests a relationship in their etiologies. PMID- 11904694 TI - Myofibroma of the upper arm in a 52-year-old woman. AB - Myofibromas are mesenchymal tumors that most often present in infancy. There are sporadic cases in adults throughout the literature. Myofibroma will demonstrate a variable appearance on CT and ultrasound, but tends to have a fairly consistent appearance on MR imaging. This report describes the MR and sonographic findings as well as the pathologic correlation of a myofibroma in the arm of a middle-aged woman. The patient was well, without evidence of recurrence 6 months following resection. PMID- 11904693 TI - Kissing periosteal chondroma and osteochondroma. AB - We present an unusual case of two synchronous chondroid matrix lesions in the proximal femur of a 16-year-old male. They were incidental findings, which subsequently underwent surgical excision and confirmed the imaging findings of a "kissing" osteochondroma and a periosteal/juxtacortical chondroma. We could not find another report of these findings in the English literature. PMID- 11904695 TI - Idiopathic widespread calcium pyrophosphate dihydrate crystal deposition disease in a young patient. AB - We report a case of generalized chondrocalcinosis in a 34-year-old man with no familial or medical history of associated diseases. Radiographs revealed calcification of the articular cartilages of the limbs, the fibrous rings around the intervertebral disks, the triangular cartilages of the wrists, and the pubic symphysis. On the basis of the presence of calcium pyrophosphate in the synovial fluid of his right ankle and generalized chondrocalcinosis, the patient was diagnosed with idiopathic calcium pyrophosphate dihydrate (CPPD) crystal deposition disease. Nonsteroidal anti-inflammatory drugs and steroid treatment provided symptomatic relief. PMID- 11904730 TI - Therapeutic effect of colon tumor cells expressing FLT-3 ligand plus systemic IL 2 in mice with syngeneic colon cancer. AB - Flt-3 ligand (FL) and interleukin-2 (IL-2) have been shown to enhance individually the antitumor response against several cancers. Therefore, treatment with a combination of FL gene-transduced tumor cell plus soluble IL-2 was studied in a murine colon adenocarcinoma model. The human full-length FL cDNA was cloned from FL-expressing cell line AML-193 by reverse transcription-polymerase chain reaction (RT-PCR). CC-36 colon tumor cells were transduced with the FL gene (CC 36-FL). In vivo and in vitro secretion of FL from CC-36-FL was confirmed by enzyme-linked immunosorbent assay (ELISA). Moreover, enhancement of dendritic cells in vivo was evaluated in mice transplanted with CC-36-FL. The therapeutic efficacy of CC-36-FL plus systemic IL-2 was tested using six groups ( n=12 13/group) of 10-week-old male Balb/c mice transplanted with 10(3) CC-36 tumor cells. Mice were treated subcutaneously with 10(6) irradiated CC-36 cells, 10(6) irradiated CC-36 cells+IL-2, 10(6) irradiated CC-36-FL cells, 10(6) irradiated CC 36-FL+IL-2, or IL-2 alone on days 4, 10 and 18 after tumor transplantation. A group of mice with no treatment served as a control. All of the treatment injections were performed subcutaneously in the left flank. IL-2 (2 x 50,000 IU) was administered intraperitoneally in 3-day cycles (days 4-6, 10-12, 17-19). Tumor growth was determined by measuring the tumor diameter. A survival experiment was performed with the same treatments, and mice were observed for survival for 100 days. The group of mice treated with the combination of CC-36 FL+IL-2 showed a significant reduction in tumor burden when compared to the no treatment group and the other control treatment groups ( P<0.05). Similarly, the group of mice treated with CC-36-FL+IL-2 displayed significant survival when compared with the other control groups (P<0.05). In Balb/c mice, the CC-36-FL plus systemic IL-2 therapy significantly decreased the tumor burden and increased the survival rate when compared to mice treated by control therapies or mice that received no treatment. PMID- 11904731 TI - Generation of functional dendritic cells for potential use in the treatment of acute lymphoblastic leukemia. AB - Immunotherapy of malignant diseases mediated by dendritic cells (DC) pulsed with tumor antigens ex vivo is a promising new tool in the individual treatment of malignant diseases. The present study focuses on the problem of how to optimize in vitro culture conditions and induce the maturation of DC with the capacity to induce antitumor immunity toward leukemic cells. DC were generated from peripheral mononuclear cells by co-cultivation with granulocyte/macrophage-colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Tumor antigens were added for 2 h after 7 days in culture. Irradiated leukemic blasts, blast lysate, apoptotic cells from the Jurkat cell line (T ALL) and their lysate were used in various concentrations for antigen pulsing. Harvested DC were phenotyped by flow cytometry, and viability was assessed using trypan blue exclusion (Annexin test). After the cells had been pulsed with tumor antigens and co-cultured with autologous lymphocytes, the production of interferon-gamma (IFN-gamma) and IL-12 was analyzed, and lymphocyte proliferative response and cytotoxicity against the target tumor cell line were assessed. The cultivation of monocytes under the described conditions led to the expression of surface markers typical of DC (i.e. CD83, CD86, HLA-DR, CD11c and CD40). Pulsation by antigens from leukemic cells further increased the cell populations expressing these markers. Antigen pulsation decreased the viability of generated DC depending on the increase in concentration of tumor antigens. Pulsed DC-lymphocyte interaction increased the proliferative response of lymphocytes and IFN-gamma production depending on the type of tumor antigens used for pulsation. The highest proliferative response was detected with DC pulsed with Jurkat cell-line lysate. Similarly to the proliferation assay, cytotoxic testing showed the highest efficiency of DC pulsed with Jurkat cell-line lysate in killing the target malignant cells. Our results show that an appropriate antigen concentration used for DC pulsing is one of the crucial factors in an effective treatment strategy, as high concentrations of tumor antigens induce apoptosis of DC, thereby rendering them non-functional. Under optimal conditions, pulsation by lysate from leukemic blasts induced the maturation of DC and led to an increase in the proliferation of autologous lymphocytes, to the production of Th1-cytokines and to the induction of cytotoxicity toward the leukemic cell line. These results are encouraging for the possible application of pulsed DC in the therapy of acute lymphoblastic leukemia. PMID- 11904732 TI - Cloning, isolation and characterization of human tumor in situ monoclonal antibodies. AB - A bacterially expressed human antibody (Ab) library (diversity approximately 10(5)) was generated from tumor-infiltrating B lymphocytes present in tissue isolated from a colon tumor. Immunoglobulin (IgG) heavy and light chain variable regions were amplified without isolating or enriching B cells, cloned into a phage-expression vector, and soluble antigen-binding fragment (Fabs) from >10(5) members of the library were screened rapidly by two distinct and complementary methodologies. In the first approach, soluble Fabs were screened by enzyme-linked immunosorbent assay (ELISA) on tumor cell monolayers. Alternatively, tumor cell surface antigens were selectively biotinylated with a plasma membrane-impermeable reagent, solubilized with non-ionic detergent, and were used to screen >10(5) members of the Ab library by capture lift. Reactive Fabs were partially characterized for tumor cell specificity and cross-reactivity, resulting in the identification of multiple Abs that bind cultured tumor cells but not normal human fibroblasts. The Fabs clustered into at least three distinct epitope specificity groups based on multiple criteria, including differential reactivity on two tumor cell lines and distinct antigen recognition patterns on western blot and immunoprecipitation. Moreover, DNA sequencing of the Ab variable regions demonstrated that the majority of the tumor-reactive Fabs were distinct and substantially different from the corresponding most homologous Ab germline gene. The relatively small size of the tumor-derived library allowed direct screening of soluble Fab of every member of the library, permitting the characterization of multiple human monoclonal antibodies (mAbs) that might not be discovered using alternative approaches, such as hybridoma technology or phage-display. PMID- 11904733 TI - Production of MCP-1 and RANTES in bladder cancer patients after bacillus Calmette Guerin immunotherapy. AB - Bacillus Calmette-Guerin (BCG) therapy induces a local immunological response mediated by cellular immune and inflammatory reactions that enhance its anti tumor efficacy in bladder cancer. Monocyte chemotactic protein-1 (MCP-1) and the "regulated on activation normal T expressed and secreted" chemokine (RANTES) are potent chemotactic molecules that attract monocytes and memory T cells. MCP-1 and RANTES levels in patients with superficial bladder cancer treated with intravesical instillations of BCG are significantly higher than in untreated cancer patients and controls. In the present study, the subjects were divided into three groups: (1) control subjects; (2) bladder cancer patients who did not receive BCG treatment; (3) bladder cancer patients who received intravesical administration of BCG. No differences in the basal production and expression of MCP-1 and RANTES mRNA were observed between BCG-treated and untreated patients. BCG treatment influenced the monocyte response to phytohemagglutinin (PHA) and BCG stimulation. After 24-h incubation, monocytes from BCG-treated bladder cancer patients released more MCP-1 and RANTES than those from untreated bladder cancer patients and controls. The anti-tumor effects of BCG observed in superficial bladder cancer therapy may depend on stimulation of the investigated chemokines, which attract monocytes/macrophages and memory T cells. PMID- 11904734 TI - Immunotherapy of solid cancer using dendritic cells pulsed with the HLA-A24 restricted peptide of carcinoembryonic antigen. AB - Carcinoembryonic antigen (CEA), an oncofetal glycoprotein overexpressed in most gastrointestinal and lung cancers, is a candidate molecule for cancer immunotherapy. Recently, a CEA-derived 9-mer peptide, CEA652 (TYACFVSNL), has been identified as the epitope of cytotoxic T lymphocytes restricted with human leukocyte antigen (HLA)-A24, which is present in 60% of the Japanese population and in some Caucasians. The authors performed a clinical study of a vaccine using autologous dendritic cells (DCs) pulsed with CEA652 and adjuvant cytokines, natural human interferon alpha (nhuIFN-alpha), and natural human tumor necrosis factor alpha (nhuTNF-alpha), for the treatment of patients with CEA-expressing advanced metastatic malignancies. Ten HLA-A24 patients with advanced digestive tract or lung cancer were enrolled in the study to assess toxicity, tolerability and immune responses to the vaccine. DCs were generated from plastic adherent monocytes of granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood mononuclear cells (PBMCs) in the presence of granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL-4). Generated DCs showing an immature phenotype were loaded with CEA652 and injected into patients intradermally and subcutaneously with 50% of the dose administered by each route every 2 weeks for a total of ten vaccinations. The total dose of administered DCs ranged from 2.7x10(7)cells to 1.6x10(8)cells. Adjuvant cytokines, i.e., 1x10(6) U/body of nhuIFN-alpha and nhuTNF-alpha, were administered to patients twice a week during the vaccination period. No severe toxicity directly attributable to the treatment was observed, and the vaccine was well tolerated. In the delayed type hypersensitivity (DTH) skin test, two patients showed a positive skin response to peptide-pulsed DCs after vaccination, although none of the patients tested positive prior to vaccination. In the two patients who showed a positive skin response disease remained stable for 6 and 9 months respectively. These results suggest that active immunization using DCs pulsed with CEA652 peptide in combination with the administration of adjuvant cytokines is a safe and feasible treatment procedure. PMID- 11904736 TI - Immune response to E7 protein of human papillomavirus type 16 anchored on the cell surface. AB - To target the E7 protein of human papilloma virus 16 to the cell surface, a fusion gene was constructed. It encodes the signal peptide, part of the immunoglobulin (IgG)-like domain, the transmembrane anchor of vaccinia virus (VV) hemagglutinin (HA), and the complete E7-coding sequence. The fusion gene was expressed under the HA late promoter by a recombinant VV, designated VV-E7-HA. The E7-HA protein was displayed on the surface of cells infected with the recombinant virus and was more stable than unmodified E7. The biological properties of the VV-E7-HA virus were compared with those of a VV-E7 virus that expressed the unmodified E7 and with a VV expressing the Sig-E7-LAMP fusion protein. While the first two of these recombinants were based on VV strain Praha, the third was derived from the WR strain of VV. Infection of mice with the VV-E7 HA virus induced the formation of E7-specific antibodies with the predominance of the IgG2a isotype, whereas the other two viruses did not induce the formation of E7-specific antibodies. Unlike the other two viruses, VV-E7-HA did not induce a response of cytotoxic T lymphocytes or Th1 cells and did not protect mice against the growth of E7-expressing tumors. Thus, VV-E7-HA induced a differently polarized immune response to the E7 protein than the other two viruses. PMID- 11904735 TI - Ocular symptoms in children treated with human-mouse chimeric anti-GD2 mAb ch14.18 for neuroblastoma. AB - Unusual ocular symptoms observed during intravenous treatment with anti disialoganglioside antibody (Ab) in children suffering from neuroblastoma were analyzed and the results reported. Within the framework of the German Collaborative Neuroblastoma Study NB97, 85 children with high-risk neuroblastoma received anti-GD2 monoclonal antibody ch14.18 intravenously. Side effects were regularly reported to the study center. Ocular symptoms were recorded in clinical detail, duration and development over time. Symptoms of a parasympathetic deficit corresponding to internal ophthalmoplegia, i.e. mydriasis and accommodation deficit, were found in 10 patients. They were uni- or bilateral, began after the termination of Ab infusion and improved or disappeared in all surviving children. They did not reappear or worsen upon repeated Ab infusions. The pathophysiology of these disorders remains poorly understood. It is concluded that during systemic treatment with the anti-GD2 antibody ch14.18, reversible symptoms of parasympathetic denervation of the eye may occur which, however, do not warrant termination of this treatment. PMID- 11904737 TI - Syndecan-1 in B lymphoid malignancies. AB - Syndecans are heparan sulfate-bearing proteoglycans that are found on the surface of most cells. Syndecan-1 is expressed predominantly on epithelia, but is also present on pre-B cells and plasma cells. The syndecans act to bind various effector molecules via their heparan sulfate chains, including both soluble and insoluble molecules within the extracellular milieu. These interactions promote cell adhesion to extracellular matrix and to adjacent cells. In addition, the syndecans can bind to and affect the biological activity of a number of heparin binding growth factors. Thus, syndecan-1 can play a dramatic role in regulating cell behavior. In this review we discuss the expression of syndecan-1 on malignant B lymphoid cells as well as specific structure-function relationships of the molecule. Emphasis is placed on the important role that syndecan-1 has in regulating the growth of B lymphoid malignancies, particularly multiple myeloma. PMID- 11904738 TI - IgM myeloma: a report of four cases. AB - IgM myeloma is a rare disease, accounting for approximately 0.5% of multiple myelomas (MM). Here we report four cases of IgM multiple myeloma. Two were diagnosed in advanced clinical stages with multiple osteolytic lesions, leading to hypercalcemia in one patient. Bone marrow morphology showed a variable degree of infiltration with mainly mature plasma cells. An immunophenotypic analysis performed in one case showed expression of CD38 and monoclonal cytoplasmatic immunoglobulin. Interphase fluorescence in situ hybridization performed in one case did not reveal any aneuploidies or deletions of the retinoblastoma, P16, or P53 tumor suppressor genes. While one patient with a smoldering IgM myeloma did not need specific therapy, the others received cytotoxic treatment based on standard chemotherapy for MM. The outcomes were one stable disease, one sustained complete remission, and one progressive disease. All four patients were alive 1 year after diagnosis. One died due to progressive disease after 31 months. We conclude that IgM myeloma shares clinical and histological features with other MM rather than with Waldenstrom's macroglobulinemia, which is most commonly diagnosed in cases with IgM monoclonal gammopathy. Since MM and Waldenstrom's macroglobulinemia differ in prognosis and treatment strategies, the two disease entities should be distinguished based on clinical criteria, bone marrow morphology, and immunophenotypic analysis. PMID- 11904739 TI - Serum thymidine kinase and soluble interleukin-2 receptor predict recurrence of malignant lymphoma. AB - Before and after therapy, serum thymidine kinase (TK) and soluble interleukin-2 receptor (sIL-2R) were serially determined in 28 patients with malignant lymphoma (ML). In 15 patients achieving and maintaining complete remission (CR) for more than 2 years, serum TK and sIL-2R were unchanged or decreased gradually. In contrast, logarithmic linear increases of TK and sIL-2R were observed in 13 relapsed patients. The increments of the serum markers occurred more than 10 months before the relapse. A significant positive correlation between the slope of the line for TK and that for sIL-2R was noted. The doubling time for TK estimated from the slope also showed a positive correlation with that for sIL-2R. Taken together, serum TK and sIL-2R were shown to be quite sensitive and interrelated serum markers for the recurrence of ML. Slopes of logarithmic linear increase, which are proper and specific for the individual patients, are inversely correlated with the doubling time and reflect proliferation of ML. We conclude that serum TK and sIL-2R are better predictors of relapse than LDH and the international prognostic index (IPI). PMID- 11904740 TI - High reliability and sensitivity of the BCR/ABL1 D-FISH test for the detection of BCR/ABL rearrangements. AB - The BCR/ABL1 fusion gene is mainly caused by the t(9; 22)(q34; q11.2) translocation, which results in the Philadelphia (Ph) chromosome. The Ph chromosome is the typical hallmark in chronic myeloid leukemia (CML), but can also be present in acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). The BCR/ABL1 rearrangement is an important tumor classification marker and a useful prognostic factor allowing an adequate therapy management. Ph chromosome detection by conventional cytogenetics (CC) can be hampered by low quantity and quality of metaphases from tumor cells. Furthermore, BCR/ABL1 rearrangements may be hidden due to cryptic rearrangements or complex aberrations. Therefore, molecular cytogenetic methods turned out to be useful tools for the detection of BCR/ABL1 rearrangements. We performed fluorescent in situ hybridization (FISH) with the recently developed BCR/ABL1 D-FISH probe (QBIOgene, Illkirch, F) on cultured bone marrow and peripheral blood cells of 71 patients with CML, ALL, AML, and myeloproliferative disorder (MPD). FISH results and the results of banding methods were directly compared. Based on the analyses of >200 nuclei per patient, D-FISH correlated closely with CC and allowed an accurate quantification of BCR/ABL1 rearrangements even in a low percentage of aberrant cells. No false positive or false-negative results were obtained. Furthermore, the D-FISH probe detected three cryptic and one complex BCR/ABL1 rearrangement, which were not visible by CC. We conclude that D-FISH reliably detects standard Ph chromosomes as well as its variant translocations and accurately quantifies BCR/ABL1 rearrangements prior and during cancer treatment as well as in the phase of remission, in daily routine tumor cytogenetic diagnostics. PMID- 11904741 TI - Parenteral iron supplementation for the treatment of iron deficiency anemia in children. AB - Iron-deficiency anemia impairs growth and intellectual development in children, which can be reversed only by early diagnosis and iron supplementation. Oral supplementation can efficiently replace stores, but in many cases parenteral iron is needed. Unfortunately some adverse reactions have limited its use in children. We compared the efficacy and safety of intramuscular and intravenous administration in 33 evaluable children with severe iron deficiency and/or iron deficiency anemia who failed to respond to oral iron supplementation. Nineteen children received intravenous infusion and 14 intramuscular injections. All children showed recovery from iron-deficiency anemia, with statistically similar improvement in hemoglobin levels. The duration of treatment was longer in those receiving intramuscular injection. Parenteral iron therapy for the treatment of iron-deficiency anemia is a rapid, easy, and definitive solution to a long troubling situation. We suggest the use of parenteral iron, in particular intravenous iron, in children who do not recover from severe iron-deficiency anemia after oral therapy. We should consider the physical and neuropsychological sequelae of long-lasting iron deficiency in children and the fact that oral supplementation is less likely to replace iron stores. PMID- 11904742 TI - Hemolytic anemia after kidney transplantation: case report and differential diagnosis. AB - A 58-year-old woman presented with hemolysis and thrombocytopenia 2 weeks after receiving a kidney graft. Hemolytic uremic syndrome was initially suspected, because in addition to hematological changes the graft function was missing. Unexpectedly, the results of the direct antiglobulin test became positive (4+), which is not normally observed in the hemolytic uremic syndrome. Differentiation of the eluted antibodies revealed anti-rhesus D specificity, which had to be interpreted either as an autoantibody of patient's origin or, hypothetically, as a "graft versus host" antibody of donor origin. Gm- and Km allotyping of these antibodies demonstrated a pattern which differed from the patient's but was identical to that of the kidney donor. Therefore hemolysis could be explained unambiguously by "graft versus host" antibodies. Whether the thrombocytopenia was also due to an immune process was not clear, although some evidence favors this hypothesis. Immunosuppressive treatment remained unchanged and several red blood cell transfusions were necessary before reactivity of the direct antiglobulin test diminished and became negative 7 weeks after kidney transplantation. The occurrence of hemolysis in the early posttransplantation period should thus draw attention to the possibility of "graft versus host" antibodies directed against red cells. Concomitant thrombocytopenia may occur. Donor screening for irregular erythrocyte antibodies should be performed whenever solid organ transplantation is intended. PMID- 11904743 TI - Successful pregnancy outcome in a patient with Gaucher's disease and antiphospholipid syndrome. AB - Gaucher's disease is characterized by increased incidence of several autoantibodies, but autoimmune phenomena are rare in Gaucher patients. We report the first occurrence of Gaucher's disease and antiphospholipid syndrome in the same patient. A 27-year-old woman with hepatosplenomegaly and thrombocytopenia who was diagnosed as having Gaucher's disease with the genotype 1226G/1226G developed Coombs'-positive hemolytic anemia, recurrent abortions, and a high titer of IgG and IgM anticardiolipin antibodies constituting the diagnosis of antiphospholipid syndrome. A successful pregnancy outcome was achieved by combined therapy with aspirin, low-molecular-weight heparin, prednisone, and enzyme replacement therapy with imiglucerase. The possible pathogenicity of antiphospholipid antibodies found in the sera of many asymptomatic Gaucher patients should be further clarified. PMID- 11904744 TI - A space-occupying lesion of the skull base, masked by nasopharyngeal lymphatic tissue hypertrophy and causing cranial nerve dysfunction in an HIV-infected patient. AB - Patients infected with HIV are at increased risk of developing lymphoma. The lymphomas often involve extranodal sites and +/-90% are of B-cell phenotype. We describe an HIV-infected patient with unilateral multiple cranial nerve dysfunction, most likely as a result of a nasopharyngeal B-cell non-Hodgkin's lymphoma in which early histologic confirmation of the diagnosis was delayed by the simultaneous presence of nasopharyngeal lymphatic tissue hypertrophy. It is of practical importance to recognize non-Hodgkin's lymphoma as a cause of cranial nerve dysfunction and to be aware of the possibility and the implications of the simultaneous presence of nasopharyngeal lymphatic tissue hypertrophy in HIV infected patients. PMID- 11904745 TI - Fatal diffuse alveolar damage complicating acute myeloid leukemia with abnormal eosinophils and trisomy X. AB - We describe a case of acute myeloid leukemia (AML) with abnormal eosinophils in a 44-year-old Chinese woman that was complicated by diffuse alveolar damage (DAD) and pulmonary hemorrhage (PH) shortly after induction chemotherapy. Cytogenetic study of bone marrow cells at diagnosis showed a rare aberration of trisomy X (+X) as the sole acquired karyotypic abnormality. We speculate that tissue damage by cellular constituents of the abnormal eosinophils that were released on cell lysis after chemotherapy might be etiologically linked to the occurrence of fatal pulmonary complications. PMID- 11904746 TI - Use of recombinant human deoxyribonuclease (DNase) for processing of a thawed umbilical cord blood transplant in a patient with relapsed acute lymphoblastic leukemia. AB - This case report describes for the first time the use of a recombinant human enzyme deoxyribonuclease (rhDNase) containing solution for the processing of a thawed umbilical cord blood (UCB) unit prior to successful transplantation to avoid cell losses by clotting phenomena. A 6-year-old boy received an unrelated 2/6 HLA antigen mismatched UCB transplant for high-risk Burkitt type acute lymphoblastic leukemia. The UCB unit was provided as a volume-reduced sample after buffy coat separation with a final volume of 36 ml. To avoid the loss of nucleated cells due to cell clumping during thawing procedure cells were washed with a solution containing the rhDNase. No visible clotting of the resuspended unit occurred, and the patient was transplanted with 2.9x10(7) nucleated cells/kg body weight without any acute or chronic side effects due to rhDNase. On day +35, PCR analysis of bone marrow aspirate showed complete chimerism, and the child engrafted with an absolute neutrophil count greater than 0.5x10(9)/l on day +47. Platelet transfusion independence was achieved on day +120. In conclusion, the supplementation of rhDNase to the washing and resuspension solutions of a thawed UCB unit is effective to prevent cell losses prior to transplantation. However, further investigations must be performed to confirm the safety of this procedure. PMID- 11904747 TI - Philadelphia-positive T-cell acute lymphoblastic leukemia with polymyositis, migratory polyarthritis and hypercalcemia following a chronic myeloid leukemia. AB - Transformation of chronic myeloid leukemia (CML) often results in acute myeloblastic or, less frequently, in precursor B-cell acute lymphoblastic leukemia (ALL). T-cell blast crisis is rare. Hypercalcemia has also been described as a rare complication of CML, but this usually occurs as a terminal event. Here we report a case of a 35-year-old woman who developed a CD4(+)/CD8(+) T-cell ALL 2 years after the diagnosis of a typical Ph(+) CML. Polymyositis and polyarthritis preceded by 4 months, and symptomatic hypercalcemia occurred just before blastic transformation, probably representing paraneoplastic manifestations of the disease. PMID- 11904748 TI - Cutaneous lesions and chronic myelocytic leukemia (CML) in progression. PMID- 11904749 TI - Genetic characterisation of granular cell tumours. AB - Seven granular cell tumours (GCT) were studied by comparative genomic hybridisation. These consisted of two cerebral and one pituitary GCT as well as four other central nervous system tumours predominantly composed of granular cells (one glioblastoma, meningioma, ganglioglioma and neurinoma each). DNA copy number changes were found in four of seven tumours. Overall, losses (mean: 1.0 per tumour) were more frequent than gains (mean: 0.6 per tumour) and no high level gains were found. The only DNA copy number change found in two tumours (both cerebral GCT) was loss of 13q21, while the other nine aberrations were only encountered once each: the granular cells of one cerebral GCT additionally showed +1p32-pter, +9q33-qter, +20q, -4, and -18q, of the glioblastoma +7 and -10, and of the meningioma -1p and -22q, while the pituitary GCT, the ganglioglioma and the neurinoma showed no imbalances. Our data suggest that a variety of genetic changes are associated with granular cell formation and support the notion that GCT are not a distinct tumour entity characterised by specific chromosomal imbalances but rather a degenerative phenomenon of cells of various origin showing DNA copy number changes akin to the underlying tumour. PMID- 11904750 TI - Disentangling the pathology of schizophrenia and paraphrenia. AB - With increasing longevity, the number of older schizophrenic patients is growing. Previous criteria used the age of symptom onset to differentiate between the late manifestations of early-onset schizophrenia and late-onset schizophreniform disorders. Current DSM-IV or ICD 10 nomenclatures do not differentiate between early- and late-onset schizophrenia. Many decades of repeated failures to provide for distinguishing neuropathological findings have prompted narrower definition criteria. Since psychotic or schizophreniform symptoms in old age may be a manifestation of Alzheimer's disease, we attempted to base a distinction between both early- and late-onset schizophrenia on the presence of degenerative changes. This study examined the brains of 64 schizophrenic patients and 18 controls immunocytochemically for tau and amyloid staining. We divided patients according to their ages at the onset of symptoms: <40, >40. Using Braak's classification, we assessed the presence of neurofibrillary pathology. Stages III and IV were observed in 11.1% (2/18) of controls, 36.7% (11/30) of early-onset schizophrenics (<40) and 58.8% (20/34) of late-onset (>40) schizophrenics (chi2=11.39, P =0.003). Stages V and VI (definite Alzheimer's disease) did not significantly differ among groups (chi2=3.6, P =0.165). Astrocytes, subependymal and fibroblastic, also exhibited tau-positive tangles. Chi-square analysis of the data revealed a significant association between tau-positive glial tangles and Braak staging ( P =0.002). Amyloid deposits were sparse in comparison to tau related changes. The restricted limbic tauopathy not only affected a majority of patients with late-onset schizophrenia (19 female: 1 male among positive cases) ( P =0.048) but also appeared in one-third of those elderly schizophrenic patients whose symptom onset occurred before 40 years of age (8 female: 3 male among positive cases) ( P =0.048). The resultant changes define a type of neuronal cytoskeletal disruption that alters the flow of information through the hippocampus and provides a useful clinico-pathological correlate to a group of patients until recently diagnosed as schizophrenic. PMID- 11904751 TI - Shaken infant syndrome: developmental neuropathology, progressive cortical dysplasia, and epilepsy. AB - This study describes the developmental neuropathology of two infants who survived 7 and 9 years, respectively, an episode of violent shaking (shaken infant syndrome) early in their lives. The shaking injuries include cortical and subcortical contusions, hemorrhages, hypoxic/ischemic and axonal damage, and severe edema. The types, distribution, and resolution of these shaking injuries are detailed by sequential radiographic studies and by pathologic examination at postmortem. Despite their severity and extent, these injuries resolved in a relatively short period of time. By 6 months, the original injuries are repaired and the resultant encephaloclastic encephalopathies (e.g., multicystic encephalomalacia, porencephaly, generalized white matter attenuation, diffuse cortical atrophy, microgyria, ulegyria, and hydrocephalus ex vacuo) are well established. No appreciable pathologic differences are detected when radiographic findings at 6 months of age are compared to postmortem observations. On the other hand, undamaged and/or partially damaged cortical regions survive the original insult and undergo post-injury reorganization that transforms the residual cortex structural and presumably functional organization. Prominent features of this post-injury reorganization include progressive cortical dysplasia with cytoarchitectural disorganization, laminar obliteration, morphologic and functional (synaptic reorganization) transformation of some neurons, preservation of layer 1 intrinsic fibers and Cajal-Retzius cells, and the presence of large (hypertrophic) intrinsic neurons with intense neurofilament immunoreactivity. We propose that this progressive dysplastic process modifies the residual cortex structural and functional organization, influences the child's neurological and psychological maturation, and may play a significant role in the pathogenesis of ensuing neurological and/or psychological sequelae. PMID- 11904752 TI - Vascular fibrosis and calcification in the hippocampus in aging, Alzheimer disease, and Down syndrome. AB - Study of the hippocampal formation of 82 subjects, including 25 control subjects from 33 to 83 years of age, 34 subjects with Alzheimer disease (AD) from 65 to 89 years of age, and 23 subjects with Down syndrome (DS) from 33 to 72 years of age, revealed hippocampal vasculopathy with fibrosis and calcification (VFC) in 40% of control, 59% of AD, and 4% of DS subjects. VFC starts in the precapillaries/capillaries in the molecular layer of the dentate gyrus (DG) and expands to the granule cell and polymorphic cell layer of the DG, and to the stratum lacunosum/molecular in the CA1 sector. Vasculopathy spreads from the tail to the body and, in a few cases, to the head of the hippocampal formation. Light and electron microscopy reveal thickening of the vascular wall with fibrosis, calcification, and enforcement of the astrocyte interface with vessels with anchorage densities associated with hemidesmosome-like structures. In moderately and severely affected cases, fragmentation and removal of calcified and occluded vessels result in local reduction of vascular network. In two AD subjects, severe vascular calcification extending from the tail to the head of the hippocampal formation was associated with loss of almost all neurons in the CA1 sector and in the subiculum proper, corresponding to hippocampal sclerosis. The topography of affected vessels and the patterns of neuronal loss reflect the middle hippocampal artery distribution with its precapillary/capillary network. The similar prevalence of vasculopathy in the AD group and in the age-matched control group, and the presence of hippocampal VFC in only one subject in the DS cohort, 96% of which is affected by Alzheimer-type pathology, oppose the link between AD and this form of vasculopathy. However, severe VFC affects the pattern of AD pathology locally by deletion of neurofibrillary degeneration and beta amyloidosis in the CA1 sector, subiculum proper, and the molecular layer of the dentate gyrus. Hippocampal VFC appears to be a form of vascular pathology with a unique predilection for the middle hippocampal artery and corresponding capillary network, which results in patchy neuronal loss in moderately affected subjects and in almost total neuronal loss in the area of impaired blood supply in severely affected subjects. These observations suggest an etiologic link between hippocampal VFC and hippocampal sclerosis. PMID- 11904753 TI - Neuropathology of two members of a German-American kindred (Family C) with late onset parkinsonism. AB - We present genealogical and longitudinal clinical observations and autopsy findings of a previously reported kindred, Family C (German-American), with late onset autosomal dominant parkinsonism with evidence for linkage on chromosome 2p13. The clinical phenotype includes the cardinal features of idiopathic Parkinson's disease. In addition, postural tremor and dementia are detected in some individuals. Two members of the kindred, one affected and one unaffected have recently come to autopsy. The unaffected family member was an 82-year-old woman whose brain showed only mild age-related pathology and no evidence of subclinical Lewy body disease. In contrast, the affected family member was an 83 year-old man whose brain had neuronal loss, gliosis and Lewy bodies in the substantia nigra and other monoaminergic brain stem nuclei, as well as the basal forebrain and amygdala. Lewy bodies and Lewy neurites had a distribution typical of cases of idiopathic Parkinson's disease. Thus, the clinical and pathological findings in this family with autosomal dominant parkinsonism are similar to those of sporadic Parkinson's disease. PMID- 11904754 TI - Microglia promote glioma migration. AB - Diffuse astrocytic gliomas extensively infiltrate brain tissue and contain numerous microglial cells, but it is unknown whether these two characteristic features are pathogenetically related. We therefore studied the effects of murine microglial cells on motility of GL261 mouse glioma cells using Boyden chamber assays. In the presence of microglia, glioma cell migration occurred earlier, and after 48 h it was threefold higher as compared to incubations without microglia. This effect was mediated by substances released from microglia, because similar effects were observed by microglia-conditioned medium, and it was specific to microglia, because oligodendroglia and endothelial cells only weakly stimulated glioma cell migration. Microglia activating substances (GM-CSF, LPS) led to a further increase of motility. These data support the notion that microglia accumulation in diffuse glial tumors does not merely represent a nonspecific reaction to tissue injury, but reflects participation of these cells in supporting and promoting the invasive phenotype of astrocytoma cells. PMID- 11904755 TI - Apoptosis in cerebrum of macular mutant mouse. AB - Neuronal cell death in the brain of macular mutant mouse, a model of copper metabolism abnormality, has features of both apoptosis and necrosis. Apoptotic cells were morphologically identified by the terminal deoxynucleotidyl transferase nick end-labeling (TUNEL) method and electron microscopy. Numerous TUNEL-positive cells were identified in the cerebral cortex, hippocampus and thalamus of the hemizygotes after postnatal day 11. Ultramicroscopic studies confirmed that a number of cells had apoptotic features characterized by condensation and segregation of the nuclei. Furthermore, genomic DNA gel electrophoresis revealed a laddering pattern in the hemizygous brain. Starvation, which produced a low body weight in normal mice similar to that seen in the hemizygotes, did not result in an increase of TUNEL-positive cells. We also found that there was no increase of apoptotic cells in the brains of heterozygotes and copper-supplemented hemizygotes. Immunocytochemical analysis revealed that the distribution of copper/zinc superoxide dismutase-containing cells differed from that of TUNEL-positive cells. These findings suggest that copper deficiency is a key factor triggering apoptosis in the brain of macular mutant mouse through a mechanism different from suppression of antioxidant action of the dismutase. The improved survival period of the copper-supplemented hemizygotes may be attributed, in part, to inhibition of excessive neuronal apoptosis identified in the late stage of the disease. PMID- 11904756 TI - Stage-dependent and sector-specific neuronal loss in hippocampus during Alzheimer's disease. AB - Recent stereological studies documented a severe loss of hippocampal neurons in end-stage Alzheimer's disease. The development of the disease, however, is progressive and slow, over clinically inconspicuous decades. The Braak-staging system distinguishes six histopathological stages some of which are not accompanied by clinical symptoms. We analyzed hippocampal cell loss in correlation to Braak stages. Neuron numbers were determined with unbiased stereological principles in a defined subportion of the hippocampus of 28 subjects. There were no age-dependent neuronal losses in any of the hippocampal subdivisions examined. Compared to stage I, pyramidal cell loss in CA1 was reduced by 33% in stage IV ( P<0.02) and by 51% in stage V ( P<0.0002). In the subiculum, considerable neuron loss was seen only in stage V (22%; P<0.09). Other subdivisions of the Ammon's horn showed no neuron loss. Neuron loss was greater than volume loss, e.g., neuron loss of 51% between stages I and V in CA1 was accompanied by volume loss of only 25%. Our findings indicate (i) that neuronal loss is sector and stage dependent, (ii) that neuronal loss in CA1 and the subiculum is related to the formation of neurofibrillary tangles, and (iii) that neuron loss makes a weak contribution to the observed volume loss. PMID- 11904757 TI - Neuron loss from the hippocampus of Alzheimer's disease exceeds extracellular neurofibrillary tangle formation. AB - Neurofibrillary tangle (NFT) formation in the CA1 region of the hippocampus is one of the early events in the pathogenesis of Alzheimer's disease (AD). As the disease progresses more NFTs form and there is substantial neuron loss. In this study we investigated whether NFT formation accounts for all the CA1 pyramidal neuron loss seen in AD. Using unbiased stereological techniques, we estimated the total number of neurons and the number of intra- and extra-cellular NFTs in the hippocampus of 10 patients with AD and 10 age-matched controls. Marked neuronal loss (approximately 60%) was identified in AD, although NFTs accounted for only a small proportion of this loss (2.2-17.2%, mean 8.1%). Analysis of NFT accumulation with duration of dementia showed a linear relationship, supporting the belief that NFTs progressively accumulate with time. PMID- 11904759 TI - Involvement of Fas/Fas ligand system in the spinal cords of HTLV-I-associated myelopathy. AB - To investigate whether the Fas/Fas ligand (FasL) system is involved in the pathogenesis of HTLV-I associated myelopathy (HAM), expression of Fas/FasL in the spinal cord lesions of HAM patients was examined by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). Fas was preferentially expressed on infiltrating T cells in active-chronic lesions of HAM. FasL expression was up-regulated on various cells, mainly microglia/macrophages in active-chronic lesions. In contrast, Fas expression was markedly down-regulated in inactive-chronic lesions of HAM patients who had a long duration of illness. The expressions of Fas/FasL in inactive-chronic lesions were comparable to those of normal controls. In normal controls, vascular endothelial cells constitutively displayed both Fas and FasL immunoreactivity, while microglia expressed FasL. RT PCR confirmed constitutive expression of both Fas mRNA and FasL mRNA in the spinal cords of HAM patients, and in controls. Our results indicate that Fas/FasL system is involved in the inflammatory process in the central nervous system of HAM patients. PMID- 11904758 TI - Morphologically distinct plaque types differentially affect dendritic structure and organisation in the early and late stages of Alzheimer's disease. AB - We have investigated the effects of the deposition of insoluble beta-amyloid plaques on dendritic morphology within the neocortex. Labelling for beta-amyloid identified three morphologically distinct plaque types present both within the brains of preclinical Alzheimer's disease (AD) and end-stage AD cases. In both preclinical and end-stage AD, the percentage area occupied by diffuse plaques contained a greater density of labelling for microtubule-associated protein-2 (MAP2) relative to the surrounding neuropil (case type, ratio of MAP2 labelling in plaque to MAP2 labelling in surrounding neuropil +/- SEM: preclinical, 1.27+/ 0.04; end-stage, 1.32+/-0.05). In contrast, there was a greater density of MAP2 labelled processes surrounding dense-cored plaques compared to that found within the plaque area (preclinical, 0.73+/-0.05; end-stage, 0.62+/-0.07). Fibrillar plaques demonstrated a transition from the early to late stages of AD, with a substantial decrease in the density of MAP2 labelling within the plaque area in end-stage AD cases relative to preclinical AD cases (preclinical, 1.01+/-0.1; end stage, 0.72+/-0.05). The morphology of dendrites associated with dense-core or fibrillar plaques suggest physical disruption of the neuropil by beta-amyloid plaque formation. These data demonstrate that plaque isoforms differentially affect dendritic morphology in both the early and late stages of AD, with progression to clinical AD associated with evolving dendritic damage localised to fibrillar and dense-core plaques. PMID- 11904760 TI - Active, phosphorylation-dependent MAP kinases, MAPK/ERK, SAPK/JNK and p38, and specific transcription factor substrates are differentially expressed following systemic administration of kainic acid to the adult rat. AB - Excitotoxicity is considered a major cell death inductor in neurodegeneration. Yet the mechanisms involved in cell death and cell survival following excitotoxic insults are poorly understood. Expression of active, phosphorylation-dependent mitogen-activated extracellular signal-regulated kinases (MAPK/ERKs), stress activated c-Jun N-terminal kinases (SAPK/JNKs) and p38 kinases, as well as their putative active, phosphorylation-dependent specific transcriptional factor substrates CREB, Elk-1, ATF-2, c-Myc and c-Jun, has been examined following systemic administration of kainic acid (KA) at convulsant doses to rats. Increased phosphorylated MAPK (MAPK(P)) immunoreactivity has been found at 3 and 6 h in the vulnerable regions entorhinal cortex and CA3, in which neurons are committed to die, as well as in sensitive regions dentate gyrus and gyrus cinguli, in which neurons will survive. JNK(P) has been observed in the entorhinal cortex and dentate gyrus, and p38(P) immunoreactivity occurs in the entorhinal cortex. Strong c-Myc(P) expression parallels MAPK(P) immunoreactivity in the entorhinal cortex, CA3, dentate gyrus and gyrus cinguli, showing that enhanced c-Myc(P) expression does not preclude cell death or cell survival. Selective decrease of CREB(P) immunoreactivity in entorhinal cortex and CA3 indicates CREB(P) reduction associated with cell death. Strong c-Jun(P) immunoreactivity has been found in the entorhinal cortex, CA3 and dentate gyrus, thus suggesting that regulation of two opposing cellular programs (cell death or cell survival) of c-Jun(P) depends on c-Jun interactions with other factors. Interestingly, ATF-2(P), and to a lesser extent Elk-1(P), is selectively increased in the dentate gyrus. These results suggest ATF-2(P) involvement in cell survival of dentate gyrus granule cells. The present results demonstrate activation of specific MAPK pathways in association with either cell death or cell survival triggered by KA. Furthermore, increased Ras activation, as seen with p21 Ras activation assay, indicates a crucial role for Ras in activating MAP kinases following excitotoxic insult. PMID- 11904761 TI - Impairment of mitochondrial DNA repair enzymes against accumulation of 8-oxo guanine in the spinal motor neurons of amyotrophic lateral sclerosis. AB - Oxidative stress plays an important role in the pathogenesis of amyotrophic lateral sclerosis (ALS). In the present study, we investigated the expression of two major human enzymes that prevent errors caused by 8-oxoguanine (8-oxoG), a mitochondrial form of 8-oxoG DNA glycosylase (hOGG1) and oxidized purine nucleoside triphosphatase (hMTH1). We also investigated the relationship between their expression and the 8-oxoG accumulation observed in the large motor neurons of the lumbar spinal cord in seven cases of adult onset sporadic ALS, four cases of subarachnoid hemorrhage (SAH) and four control cases. 8-oxoG immunoreactivity increased in most large motor neurons in both the ALS and SAH cases. However, the large motor neurons in the control cases often lacked hOGG1 immunoreactivity, although some neurons expressed hOGG1 in either homogeneous or fine granular patterns. In SAH cases, most large motor neurons showed a fine granular pattern proportional to the increased 8-oxoG immunoreactivity. However, only half of the remaining motor neurons in ALS expressed hOGG1 in the fine granular pattern, and the rest did not show any immunoreactivity. In addition, small aggregates of hMTH1 in the nuclei of the anterior horn cells were present in several ALS cases. Our results indicate that the oxidative damage accumulates in the mitochondria of motor neurons in ALS, and that hOGG1 does not repair the damage efficiently, which may lead to a loss of motor neurons in ALS. PMID- 11904762 TI - Loss of heterozygosity on chromosome 22 in human ependymomas. AB - Ependymomas are glial tumors of the brain and spinal cord. The most frequent genetic change in sporadic ependymomas is monosomy 22, suggesting the presence of an ependymoma tumor suppressor gene on chromosome 22. Thirty-three pairs of matched normal and tumor specimens from ependymoma patients were genotyped using 12 polymorphic microsatellite markers spanning the long arm of chromosome 22. Allelic deletion was found in 12 of 33 tumors (36.4%). Eight tumors showed partial deletions and 4 tumors exhibited loss of the entire arm of 22q. We identified two common regions of deletion: one at 22q11.21-12.2 flanked by markers D22S420 and D22S300, and a second candidate region at 22q13.1-13.3 between D22S274 and D22S1149. The size of each region was 21.1 and 2.4 cM, respectively. Thus, our results suggest that one or more tumor suppressor genes associated with ependymoma may be present on chromosome 22. Comparison of these results with clinicopathological data indicate that allelic losses on 22q tend to occur more frequently in intracranial anaplastic ependymomas in children and intraspinal ependymomas in adults. PMID- 11904763 TI - Localisation of the high-affinity choline transporter-1 in the rat skeletal motor unit. AB - The rate-limiting step in neuronal acetylcholine (ACh) synthesis is the uptake of choline via a high-affinity transporter. We have generated antisera against the recently identified transporter CHT1 to investigate its distribution in rat motor neurons and skeletal muscle and have used these antisera in combination with (1) antisera against the vesicular acetylcholine transporter (VAChT) to identify cholinergic synapses and (2) Alexa-488-labelled alpha-bungarotoxin to identify motor endplates. In the motor unit, immunohistochemistry and RT-PCR have demonstrated that CHT1 is restricted to motoneurons and absent from the non neuronal ACh-synthesizing elements, e.g. skeletal muscle fibres. In addition, CHT1 is also present in parasympathetic neurons of the tongue, as evidenced by immunohistochemistry and RT-PCR. CHT1 immunoreativity is principally found at all segments (perikaryon, dendrites, axon) of the motoneuron but is enriched at neuro neuronal and neuro-muscular synapses. This preferential localisation matches well with its anticipated pivotal role in synaptic transmitter recycling and synthesis. PMID- 11904765 TI - Extraepithelial cells expressing distinct olfactory receptors are associated with axons of sensory cells with the same receptor type. AB - During critical phases of mouse development, axons from olfactory sensory neurons grow out of the nasal neuroepithelium and navigate through the connective mesenchyme tissue towards their targets in the developing telencephalic vesicle. Between embryonic days E11 and E16, populations of cells are located in the mesenchyme which express distinct olfactory receptor genes along with the olfactory marker protein (OMP); thus they express markers characteristic for mature olfactory sensory neurons. These extraepithelial cells are positioned along the axon tracts, and each population expressing a given receptor gene is specifically associated with the axons of those olfactory sensory neurons with the same receptor type. The data suggest that they either might be guide posts for the outgrowing axons or migrate along the axons into the brain. PMID- 11904764 TI - Impaired cutaneous wound healing after sensory denervation in developing rats: effects on cell proliferation and apoptosis. AB - The role of sensory nociceptor nerves in cutaneous wound healing was investigated following full-thickness 4-mm diameter dorsal cutaneous excision wounding of rats on postnatal day 12. In rats with intact innervation, wounds at 3 days contained large numbers of TUNEL- and BRDU-labeled nuclei, consistent with inflammatory cell death and granulation cell proliferation. Wound area and volume decreased through 11 days in concert with a transient appearance of alpha-smooth muscle actin-immunoreactive myofibroblasts, declining rates of cell division, and increased occurrence of apoptotic cells. Sensory denervation by capsaicin injections on postnatal days 2 and 9 reduced calcitonin gene-related peptide immunoreactive wound innervation persistently by up to 43%. This was associated with increased wound surface area and volume, and delays in scab loss and re epithelialization. Relative to control wounds, granulation tissue showed increased myofibroblast content at 5-7 days. Capsaicin-treated rats had more BRDU labeled cells, including myofibroblasts, through day 7. Numbers of TUNEL apoptotic cells per unit area of tissue section were reduced by denervation in both early and late stages of healing. We conclude that partial loss of sensory innervation impairs cutaneous wound healing in developing rats, as manifested by delayed re-epithelialization and failure of the wound area to decrease normally through at least 21 days. This is associated with an abnormally enlarged wound tissue volume resulting from increased granulation cell proliferation without proportionate increases in apoptosis. These findings suggest that nociceptor innervation plays a critical role in wound healing by regulating wound cellularity. PMID- 11904766 TI - Experimental angiogenesis of arterial vasa vasorum. AB - Vasa vasorum are important sources of oxygen and nutrients to vascular tissues and their proliferation influences the pathogenesis of arterial disease; however, the regulation of their growth is poorly understood partly because of a lack of appropriate experimental models. We cuffed common carotid arteries of rabbits with segments of the contralateral carotid artery, a procedure that resulted in rapid and extensive elaboration of adventitial vasa vasorum and connective tissue. Endothelium-lined microvessels were observed at 1 week but vessels as large as 300 microm with an organizing media were common by 3 weeks. These vasa vasorum arose primarily from the vascular supply to contiguous tissues, but also from the carotid artery. This angiogenesis was accompanied by increased expression of the angiogenic factor, vascular endothelial growth factor (VEGF), in the invading connective tissue cells and increased expression of the transcriptional regulator of VEGF, hypoxia-inducible factor-1alpha (HIF-1alpha), in these connective tissues and in the cuffing artery. These findings are consistent with the hypothesis that upregulation of HIF-1alpha and VEGF expression drives angiogenesis of vasa vasorum in this model. This simple model may be amenable to the study of the development and elaboration of vasa vasorum, especially in the context of vascular pathologies. PMID- 11904767 TI - Quantitative analysis of gap-junctional intercellular communication in precision cut mouse liver slices. AB - Direct intercellular communication through gap junction channels is involved in the maintenance of tissue homeostasis and suppression of carcinogenesis. Gap junctional communication is often altered in tumor cells but it can also be modulated in response to tumor promotors or inflammatory signals. In order to evaluate the effect of nongenotoxic compounds, suggested to be involved in tumor promotion, on gap junctional intercellular communication in the liver, we have developed a direct dye transfer method. The fluorescent dye Alexa Fluor 488 was iontophoretically injected into hepatocytes of freshly prepared, precision-cut mouse liver slices (250 microm). The area of dye spreading was monitored and quantified by microscopy. Comparison of dye spreading in connexin-32-deficient versus wild-type liver revealed a 96% decrease in connexin-32-deficient tissue. Induction of an acute phase response in connexin-32-deficient mice by intraperitoneal injection of lipopolysaccharide increased dye coupling by 33%, probably due to upregulation of connexin-26-containing gap junction channels. PMID- 11904769 TI - Localization of caveolin-3 in the sinus endothelial cells of the rat spleen. AB - The localization of caveolins in the sinus endothelial cells of the rat spleen has been demonstrated by confocal laser scanning and electron microscopy. Caveolin-3, a muscle-specific caveolin, was detected by Western blot analysis and immunofluorescence microscopy of isolated sinus endothelial cells and tissue cryosections of the spleen. During the immunofluorescence microscopy of isolated endothelial cells, both caveolin-3 and caveolin-1 were found. In tissue cryosections of the spleen, caveolin-3, as well as caveolin-1 and -2, was present in the contours and cytoplasm of the cells. Immunogold electron microscopy of tissue cryosections revealed caveolin-3, -1, and -2 to be present in caveolae in the apical, lateral, and basal plasma membranes and some vesicular profiles in the cytoplasm of sinus endothelial cells. Furthermore, caveolin-3 was colocalized with caveolin-1 in the same caveolae in the apical, lateral, and basal plasma membranes. Stress fibers and tubulovesicular structures were situated in the vicinity of caveolae labeled with anti-caveolin-3, anti-caveolin-1, and anti caveolin-2 antibodies. It is speculated that caveolae in sinus endothelial cells play an important role in the constriction of stress fibers. PMID- 11904768 TI - Porcine mesenchymal stem cells. Induction of distinct mesenchymal cell lineages. AB - The potential of mesenchymal stem and progenitor cells (MSC) to replicate undifferentiated and to mature into distinct mesenchymal tissues suggests these cells as an attractive source for tissue engineering. The objective was to establish a protocol for the isolation of porcine MSC from bone marrow and to demonstrate their ex vivo differentiation into various mesenchymal tissue cells. MSC from passage 2 were selected for differentiation analysis. Differentiation along the osteogenic lineage was documented by deposition of calcium, visualization of alkaline phosphatase activity, and by analysis of osteogenic marker genes. Adipocytes were identified morphologically and by gene-expression analysis. Deposition of type II collagen and histological staining of proteoglycan indicated chondrogenic differentiation. Therefore, porcine MSC may be introduced as a valuable model system with which to study the mesenchymal lineages for basic research and tissue engineering. PMID- 11904770 TI - An in vivo model of prostate carcinoma growth and invasion in bone. AB - Prostatic carcinoma affects 1 in 11 men and targets bone with sclerotic metastases. The study of prostate carcinoma growth in bone has been hampered by the lack of suitable animal models. We have developed an in vivo model of prostate carcinoma growth in bone by inoculating three human prostate carcinoma cell lines (PC-3, DU-145, and LNCaP) into the tibia of congenitally athymic mice. Developing tumors were analyzed by radiographic, histologic, immunohistochemical, and in situ hybridization examination. Seven of the nine PC-3 inoculated mice and all (9/9) of the DU-145 inoculated mice developed tumors in the injected limb. In contrast, inoculation with LNCaP cells failed to produce tumors (0/9). Radiologically, the tumors had a mixed sclerotic/lytic appearance with extracortical extension. All the PC-3 tumors invaded the bone marrow cavity, cortical bone, and surrounding soft tissue. The DU-145 tumors were confined to the bone marrow cavity in 7/9 animals. CK18 and Ki67 localization identified the human tumor cells and their proliferative activity, respectively. The PC-3- and DU-145-induced tibial tumors expressed alpha(1)I procollagen and osteopontin mRNA, to varying degrees. All the tumors demonstrated an up-regulation of osteoclasts at the bone/tumor interface compared with the control limbs. Thus, this is a reliable and reproducible in vivo model of prostate carcinoma growth in bone enabling the study of the interactions that occur between prostate cancer cells and bone at an important part of the metastatic cascade, namely, growth and invasion at a distant site. PMID- 11904771 TI - Cloning of a complementary DNA encoding the unique dendritic cell antigen Ki-M9. AB - Human sinus-lining cells (SLC) of the lymph node sinuses most probably represent accessory cells for the primary humoral immune response and have been shown to express a unique antigen recognized by the monoclonal antibody Ki-M9. To characterize this SLC-specific antigen further, a spleen cDNA library established in the expression vector lambda gt-11 was searched immunochemically for clones expressing the Ki-M9 antigen. Two recombinant phage clones revealed a cDNA with an open reading frame of 1666 bp encoding a 68-kDa protein. When fused with an expression vector that codes for the bacterial maltose-binding protein (MBP), the purified MBP-Ki-M9 fusion protein could be clearly detected by Western blot analysis. Furthermore, in situ hybridization with Ki-M9 cDNA as a probe confirmed the SLC-specific expression of the cloned cDNA. Additionally, Ki-M9 mRNA transcripts were detected in follicular dendritic reticulum cells of the secondary germinal centers, in a few cells of the perifollicular zone of the spleen, in some sinusoidal cells of liver, and in thymic reticular cells. A sequence database research revealed a strong homology to a murine cDNA. By applying non-radioactive in situ hybridization on mouse tissue, strong expression in SLC of the lymph node and metallophilic cells of the spleen in mouse tissue could be seen indicating that the Ki-M9 cDNA is highly conserved in the two species. Further computer analysis of the deduced amino acid sequence of the Ki M9 antigen showed a large number of potential glycosylation sites and a PEST motive, which are characteristic for rapidly degraded membrane bound proteins and represent prerequisites for the function of this cell system in initiating the humoral immune response. PMID- 11904772 TI - Ectopic mineralization of articular cartilage in the bullfrog Rana catesbeiana and its possible involvement in bone closure. AB - Mineralization of the articular cartilage is a pathological condition associated with age and certain joint diseases in humans and other mammals. In this work, we describe a physiological process of articular cartilage mineralization in bullfrogs. Articular cartilage of the proximal and distal ends of the femur and of the proximal end of the tibia-fibula was studied in animals of different ages. Mineralization of the articular cartilage was detected in animals at 1 month post transformation. This mineralization, which appeared before the hypertrophic cartilage showed any calcium deposition, began at a restricted site in the lateral expansion of the cartilage and then progressed to other areas of the epiphyseal cartilage. Mineralized structures were identified by von Kossa's staining and by in vivo incorporation of calcein green. Element analysis showed that calcium crystals consisted of poorly crystalline hydroxyapatite. Mineralized matrix was initially spherical structures that generally coalesced after a certain size to occupy larger areas of the cartilage. Alkaline phosphatase activity was detected at the plasma membrane of nearby chondrocytes and in extracellular matrix. Apoptosis was detected by the TUNEL (TDT-mediated dUTP biotin nick end-labeling) reaction in some articular chondrocytes from mineralized areas. The area occupied by calcium crystals increased significantly in older animals, especially in areas under compression. Ultrastructural analyses showed clusters of needle-like crystals in the extracellular matrix around the chondrocytes and large blocks of mineralized matrix. In 4-year-old animals, some lamellar bone (containing bone marrow) occurred in the same area as articular cartilage mineralization. These results show that the articular cartilage of R. catesbeiana undergoes precocious and progressive mineralization that is apparently stimulated by compressive forces. We suggest that this mineralization is involved in the closure of bone extremities, since mineralization appears to precede the formation of a rudimentary secondary center of ossification in older animals. PMID- 11904773 TI - The developmental potential of the inner cell mass of blastocysts that were derived from mouse ES cells using nuclear transfer technology. AB - The present study examined the causes of the low developmental potential of enucleated oocytes that have received ES cells and consequent postnatal death of the young. The inner cell masses (ICM) of nuclear-transferred blastocysts or diploid blastocysts were injected into tetraploid blastocysts (group B) or nuclear-transferred tetraploid blastocysts (group C), respectively. The developmental potential of these groups was compared with tetraploid blastocysts injected with ICM of diploid blastocysts (group A). The potential of reconstituted blastocysts to develop into live young in group B increased slightly (5%) but was significantly lower than that in group A (45%). The rate of postnatal death of young in group B did not decrease. The implantation rate of reconstituted blastocysts in group C was very low and no live fetuses were obtained. The results of the present study indicate that the inferior potential of both ICM and trophectoderm cells of nuclear-transferred blastocysts underlies the low developmental rate of nuclear-transferred oocytes receiving ES cells and the higher rate of postnatal death of ES cell-derived young. PMID- 11904774 TI - Ommatidial heterogeneity in the compound eye of the male small white butterfly, Pieris rapae crucivora. AB - The ommatidia in the ventral two-thirds of the compound eye of male Pieris rapae crucivora are not uniform. Each ommatidium contains nine photoreceptor cells. Four cells (R1-4) form the distal two-thirds of the rhabdom, four cells (R5-8) approximately occupy the proximal one-third of the rhabdom, and the ninth cell (R9) takes up a minor basal part of the rhabdom. The R5-8 photoreceptor cells contain clusters of reddish pigment adjacent to the rhabdom. From the position of the pigment clusters, three types of ommatidia can be identified: the trapezoidal (type I), square (type II), and rectangular type (type III). Microspectrophotometry with an epi-illumination microscope has revealed that the reflectance spectra of type I and type III ommatidia peak at 635 nm and those of type II ommatidia peak at 675 nm. The bandwith of the reflectance spectra is 40 50 nm. Type II ommatidia strongly fluoresce under ultra-violet and violet epi illumination. The three types of ommatidia are randomly distributed. The ommatidial heterogeneity is presumably crucial for color discrimination. PMID- 11904776 TI - Complementary distribution of NADPH-diaphorase and l-arginine in the snail nervous system. AB - Since the interneuronal messenger nitric oxide (NO) can not be stored in neurones, the regulation of the NO-producing enzyme nitric oxide synthase (NOS) is crucial. Neuronal NOS metabolises L-arginine to nitric oxide (NO) and L citrulline in a Ca(2+)-dependent manner. Thus, availability of L-arginine to NOS may modulate NO production. In this study, we examined the cellular distribution of reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase, L arginine and L-citrulline. Using NADPH-diaphorase histochemistry to visualise putative NO-producing cells and immunocytochemistry to localise L-arginine, we showed that the distribution of L-arginine-immunoreactive neurones correlates well with those of NADPH-diaphorase-positive neurones in cerebral ganglia of the pulmonate Helix pomatia. However, substrate and enzyme were visualised in separate but adjacent neurones. We further examined whether NADPH-diaphorase labelled cells contain the L-citrulline. Following elevation of intracellular Ca(2+) by the Ca(2+) ionophore, ionomycin, or by a high-K(+) solution, the number of L-citrulline-immunoreactive neurones in mesocerebrum and pedal lobe increased up to tenfold. Preincubation of ganglia with the NOS inhibitor N(G)-nitro-L arginine prevented ionomycin or high-K(+) solution-induced L-citrulline synthesis. Most L-citrulline-immunoreactive neurones contain NADPH-diaphorase activity. In conclusion, these experiments indicate a complementary distribution of NOS and L-arginine and suggest an unknown signalling pathway between neurones to maintain L-arginine and NO homeostasis. PMID- 11904775 TI - Novel genes expressed in subsets of chemosensory sensilla on the front legs of male Drosophila melanogaster. AB - Detection of courtship-activating female pheromones by contact chemoreceptors on the front legs of male Drosophila melanogaster is thought to play an important role in triggering courtship behavior. However, the chemosensory organs, cells, and molecules responsible are not known. We have isolated two genes, CheA29a and CheB42a, expressed in nonneuronal auxiliary cells within two nested subsets of chemosensory sensilla on the front legs of sexually mature, adult males. The proteins encoded by the CheA29a and CheB42a genes have no sequence similarity to each other or any other known protein, but they belong to two novel families of proteins encoded by the D. melanogaster genome. Members of the two families are predicted to have a single transmembrane domain at their amino terminus, probably to serve as a signal peptide, suggesting that they are soluble and secreted. Finally, in addition to CheA29a and CheB42a, other genes within each family are expressed preferentially in appendages where chemosensory organs are concentrated, in several cases in a male-specific manner. Our data suggest that CheA29a and CheB42a and other members of these two protein families are involved in male-specific chemical senses, perhaps pheromone response. PMID- 11904777 TI - Dynamics of apoptosis in the ovarian follicle cells during the late stages of Drosophila oogenesis. AB - In the present study, we demonstrate the apoptotic events of the ovarian follicle cells during the late stages of oogenesis in Drosophila melanogaster. Follicle cell morphology appears normal from stage 10 up to stage 14, exhibiting a euchromatic nucleus and a well-organized cytoplasm. First signs of apoptosis appear at the anterior pole of the egg chamber at stage 14A. They are characterized by loss of microvilli at the apical cell membrane, alterations in nuclear morphology, such as chromatin condensation and convolution of the nuclear membrane, and also by condensation and vacuolization of the cytoplasm. During the following stage 14B, the follicle cell nuclei contain fragmented DNA as is demonstrated by acridine orange staining and TUNEL (TdT-mediated dUTP nick end labeling) assay. Finally, the apoptotic follicle cells seem to detach from the eggshell when the mature egg chamber exits the ovariole. The detached follicle cells exhibit condensed nuclear chromatin, a disorganized cytoplasm with crowded organelles and are surrounded by epithelial cells. The above results seem to be associated with the abundant phagocytosis that we observed at the entry of the lateral oviducts, where the epithelial cells contain apoptotic cell bodies. Additionally, we tested the effect of etoposide treatment in the follicular epithelium and found that it induces apoptosis in a stage- and site-specific manner. These observations suggest a possible method of absorption of the apoptotic follicle cells that prevents the blockage of the ovarioles and helps the regular production of mature eggs. PMID- 11904778 TI - Structural definition of the neuromuscular system in the swimming-paddle opener muscle of blue crabs. AB - Blue crabs are excellent swimmers, using their highly modified last pereiopods as sculling paddles. Hence, the hypertrophied paddle opener muscle was examined for adaptations of its motor innervation by an excitor and a specific inhibitor axon. The muscle has a uniform composition of slow fibers with long (6-12 microm) sarcomere lengths. Individual fibers are richly innervated with approximately two thirds excitatory and one-third inhibitory innervation. The profuse excitatory innervation reflects the high activity levels of this motoneuron in swimming. Adaptation to sustained activity associated with swimming is also reflected in the motor nerve terminals by a high concentration of energy source, which is equally divided between glycogen granules and mitochondria, the former providing a more rapid source of energy. The excitor axon makes predominantly neuromuscular synapses, but also a few synapses onto the inhibitor axon. The location of these excitatory axoaxonal synapses suggests regional modulation of the inhibitor axon. The specific inhibitor axon makes less than two-thirds of its synapses with the muscle fiber, regulating contraction via postsynaptic inhibition. The remaining inhibitory synapses are onto the excitor axon, signaling very strong presynaptic inhibition. Such presynaptic inhibition will effectively decouple the opener muscle from the stretcher muscle even though both are innervated by a single excitor axon. PMID- 11904781 TI - Febrile neutropenia, colony-stimulating factors and therapy: time for a new methodology? AB - The utilization of granulocyte colony-stimulating factors (G-CSF) in febrile neutropenia has been controversial for many years. Berghmann et al.'s meta analysis again demonstrates that G-CSF does not have an impact on mortality in febrile neutropenia, because the depth and duration of neutropenia in the trials are variable. Also, with mortality from febrile neutropenia less than 15%, any further study would require a vast number of patients to demonstrate a difference in mortality. The Elting and Cantor review provides a new paradigm to studies in patients with febrile neutropenia. These authors recognize that cost, quality of life, life-years gained and adverse events experienced with new therapies should be evaluated, in addition to the standard measures of infection resolution and related mortality. Therefore, for the evaluation of new therapeutic interventions, a consensus on stratified risk factors or the use of an already established model could provide end-points with comparable measurements. PMID- 11904779 TI - Expression of NeuroD in the mouse taste buds. AB - NeuroD, a basic helix-loop-helix transcription factor, has been shown to play a role in the differentiation of neurons, olfactory cells, and neuroendocrine tissues. Since the taste buds have characteristics of typical paraneurons, we examined the expression of NeuroD in the taste buds of mice. By RT-PCR analysis, NeuroD mRNA was found to be expressed in the epithelium of circumvallate papillae containing taste buds, but not in the lingual epithelium lacking them. NeuroD immunoreactivity was detected in a subset of taste bud cells in the circumvallate, foliate, and fungiform papillae and in the soft palate. NeuroD expressing cells had a spindle-like shape, first appeared at postnatal day 3, and increased in number during postnatal development. After bisection of the glossopharyngeal nerves, NeuroD-expressing cells decreased in number at day 4 and disappeared from the trench wall of the circumvallate papillae by day 14. A few NeuroD-expressing taste buds reappeared at postoperative day 28. Denervation and regeneration experiments showed that expression of NeuroD in the taste buds was dependent upon gustatory innervation. Double immunolabeling with gustducin or with neural cell adhesion molecule (NCAM) showed that NeuroD-expressing cells did not express NCAM, but did express gustducin. These results suggest that NeuroD is expressed in a mature cell type, type-II cells, but not in type-III cells. PMID- 11904782 TI - Therapeutic use of granulocyte and granulocyte-macrophage colony-stimulating factors in febrile neutropenic cancer patients. A systematic review of the literature with meta-analysis. AB - The effectiveness of granulocyte and granulocyte-macrophage colony-stimulating factor (G-CSF and GM-CSF) in the treatment of febrile neutropenic cancer patients remains controversial. To assess their role in this condition, we conducted a systematic review of randomised trials published as full papers. A methodological evaluation using a specifically designed quality scale was performed before meta analysis. Eleven trials were eligible, 8 of which were meta-analysable. The median quality score for the 11 pooled trials was 58.3% (range: 33.3%-68.8%). No significant quality difference was observed between positive (colony-stimulating factor more effective) and negative trials ( P=0.36). No quality difference was observed between the 8 meta-analysable studies and the 3 others, with respective median scores of 59.3% and 50%. No advantage was detected for the use of CSF in terms of mortality from febrile neutropenia, with a relative risk of 0.71 (95% CI 0.44-1.15). The relative risk was 0.66 (95% CI 0.39-1.13) in the G-CSF subgroup and 0.97 (95% CI 0.34-2.79) in the GM-CSF subgroup. Aggregation of the results on infection-related mortality, length of stay in hospital, fever and of neutropenia duration, antibiotic therapy adaptation and duration, superinfection rate and toxicity was not possible owing to the lack of adequate data in the publications. On the basis of this review, we cannot recommend the routine use of G-CSF or GM CSF in established febrile neutropenia. PMID- 11904783 TI - Outcomes and costs of febrile neutropenia: adventures in the science and art of treatment choices. AB - The choice of therapy for febrile neutropenia is complex, because of the large number of options that are similar in safety and efficacy. However, there are a number of outcomes that may be useful when these choices have to be made. It is generally agreed that infection-related mortality is too rare an event, with the availability of modern antibiotics, to be of general use in treatment choices. Response to initial therapy may be useful, but differences among regimens in recent randomized trials only occasionally reach statistical significance, despite adequate power and sample size. The time to clinical response has been shown to vary significantly among otherwise similar regimens and may be very useful when response-based choices are made. Ideally, clinical and policy decisions should be based on a combined evaluation of outcomes and cost. In the case where clinical outcomes are the same for more than one regimen, cost minimization analysis is appropriate. In the case where clinical outcomes differ, cost-effectiveness or cost-utility is an appropriate measure on which to base decisions. The cost of therapy can be easily estimated by using the number and average cost of days of hospitalization as a surrogate. A decision-analytic model for febrile neutropenia is proposed. PMID- 11904784 TI - Challenges and options in the management of viral infections after stem cell transplantation. AB - During the period of profound combined immunodeficiency after bone marrow or peripheral blood stem cell transplantation (SCT), patients are at increased risk for serious viral disease. Recent advances in rapid diagnostic methods and the introduction of potent antiviral compounds have made it possible to establish efficient management strategies for several herpesviruses. Acyclovir, valaciclovir, and famciclovir are widely used for the treatment of herpes simplex virus or varicella zoster virus disease. Intravenous ganciclovir, foscarnet, and cidofovir are available for prevention or therapy of cytomegalovirus disease, and oral valganciclovir could become a valuable alternative to intravenous treatment if shown to be effective and safe after SCT. Preliminary data on pleconaril for therapy of picornaviral disease are promising. Future investigations may help to clarify the role of the neuraminidase inhibitors zanamivir and oseltamivir in the management of influenza in SCT recipients. The emergence of viruses resistant to antiviral drugs is of concern, and alternative treatment strategies need to be defined. PMID- 11904785 TI - Respiratory problems in advanced cancer. AB - Respiratory problems are an important issue in the palliative care setting, not only from a diagnostic and therapeutic point of view but also from emotion related aspects involving both the patients and their families and also caregivers. In this paper we consider some of the most common respiratory problems, such as dyspnea, infections, hemoptysis, hiccup. A review of the literature was performed with reference to the frequency, diagnosis and management of the above respiratory problems in patients in advanced and terminal stages of the disease. Particular emphasis was given to the importance of communication with the patients and their families, which is considered a crucial point in the care and the cure of such patients. PMID- 11904786 TI - Consultation audio-tapes: an information aid, and a quality assurance and research tool. AB - Facilities for audio-taping medical consultations are widely available, and an increasing proportion of cancer patients request and/or appreciate the offer of an audio-tape after an oncology consultation. We have examined the usefulness to patients and their families of audio-tapes of initial and follow-up consultations. We have used consultation audio-tapes as a research tool investigating doctor-patient communication. Audio-tapes may ultimately find a place as an audit tool to monitor oncologist performance. We have audio-taped several hundred medical and radiation oncology consultations over the past 10 years. We have developed methods of analysing oncologist-patient communication and studied a variety of interventions designed to enhance communication in the cancer consultation. We have offered consultation audio-tapes to cancer patients after their initial oncology consultation and after routine outpatient follow-up appointments, and documented their use. Cancer patients and their family's value consultation audio-tapes as an information aid, and to assist recall. Consultation audio-tapes document information provision and allow analysis of oncologist-patient interaction in discussion of prognosis, treatment options and seeking/giving informed consent. Consultation audio-tapes have a role in providing feedback concerning oncologist performance. Oncologists should consider installing audio-tape-recording facilities and offer new and follow-up patients a taped consultation. Audio-tapes are a valuable tool in the investigation of oncologist-patient interactions. PMID- 11904787 TI - Quality of life of breast cancer patients receiving high-dose-intensity chemotherapy: impact of length of cycles. AB - This study was designed to measure treatment side-effects and quality of life (QL) of 47 nonmetastatic breast cancer patients subjected to a dose-intensity increase while receiving a sequential high dose chemotherapy (doxorubicin+cyclophosphamide - 4 cycles). The dose-intensity increase was obtained by shortening the length of cycles from 21 to 14 days. Treatment side effects were self-assessed in terms of frequency and associated distress in cycles 1 and 3 by using a specific side-effect self-report questionnaire (19 items). Multidimensional QL measurement was performed at inclusion and before the start of cycles 2 and 4, by using the EORTC QLQ-C30. Pain was evaluated by patients on a visual analogue scale at the same times as QL evaluation. Patients' self-ratings indicated that the total number of symptoms, the number of symptoms rated by patients as quite or very distressing, and symptom frequency were comparable whatever the length of cycle. Overall, although underestimating most patients' symptoms, physicians' reports provided similar results. However, analysis of multidimensional QL showed that, in comparison to standard administration of 4 cycles of 21 days, there was a more significant deterioration of the QLQ-C30 global QL score ( P=0.01) at the second cycle of chemotherapy and of the physical functioning score ( P=0.02) at the fourth cycle when the cycle length was reduced. This study, although limited by a small patient cohort, has shown that shortening cycles to increase dose intensity had relatively few consequences on adverse treatment effects but a highly negative impact on patients' quality of life. PMID- 11904788 TI - A double-blind, randomised, parallel group, multinational, multicentre study comparing a single dose of ondansetron 24 mg p.o. with placebo and metoclopramide 10 mg t.d.s. p.o. in the treatment of opioid-induced nausea and emesis in cancer patients. AB - Nausea and emesis are common side effects of opioid drugs administered for pain relief in cancer patients. The aim of this study was to compare the anti-emetic efficacy and safety of ondansetron, placebo and metoclopramide in the treatment of opioid-induced nausea and emesis (OIE) in cancer patients. This was a multinational, multicentre, double-blind, parallel group study in which cancer patients who were receiving a full opioid agonist for cancer pain were randomised to receive one of oral ondansetron 24 mg once daily, metoclopramide 10 mg three times daily, or placebo. Study medication was started only if the patient experienced nausea and/or emesis following opioid administration. Efficacy and safety assessments were made over a study period of 24 h from the time of the first dose of anti-emetics/placebo. The study was terminated prematurely because of the difficulties in recruiting patients satisfying the stringent entry criteria. Ninety-two patients were included in the intent-to-treat population: 30 patients received placebo, 29 patients ondansetron and 33 patients metoclopramide. There was no statistically significant difference between the groups in the proportion achieving complete control of emesis (33% of patients on placebo, 48% on ondansetron and 52% on metoclopramide) or complete control of nausea (23% of patients on placebo, 17% on ondansetron and 36% on metoclopramide). Rescue anti-emetics were required in 8 of 33 patients on metoclopramide, 4 of 29 on ondansetron, and 3 of 30 on placebo. The incidence of adverse events was very low and similar in all treatment groups. Neither ondansetron 24 mg once daily nor metoclopromide 10 mg t.d.s. given orally was significantly more effective than placebo in the control of OIE in cancer patients. PMID- 11904789 TI - The effectiveness of progressive muscle relaxation training in managing chemotherapy-induced nausea and vomiting in Chinese breast cancer patients: a randomised controlled trial. AB - This study was a randomised controlled trial designed to assess the effectiveness of progressive muscle relaxation training (PMRT) in the clinical management of chemotherapy-related nausea and vomiting as an adjuvant intervention to accompany pharmacological antiemetic treatment (metoclopramide and dexamethasone i.v.). Seventy-one chemotherapy-naive breast cancer patients of an outpatient oncology unit of a university hospital in Hong Kong participated, with 38 subjects randomised to the experimental group and 33 to the control group. The intervention included the use of PMRT 1 h before chemotherapy was administered and daily thereafter for another 5 days (for a total of six PMRT sessions). Each session lasted for 25 min and was followed by 5 min of imagery techniques. The instruments used for data collection included the Chinese versions of the Profile of Mood States and the State-Trait Anxiety Inventory (measured before chemotherapy and then at day 7 and day 14 after chemotherapy), and the Morrow Assessment of Nausea and Vomiting Scale, which was used daily for the first 7 post-chemotherapy days. The use of PMRT considerably decreased the duration of nausea and vomiting in the experimental group compared with the control group ( P<0.05), whereas there were trends toward a lower frequency of nausea and vomiting ( P=0.07 and P=0.08 respectively). Neither nausea nor vomiting differed in intensity between the two groups. The significant effects were mainly evident on the first 4 post-chemotherapy days, when differences were statistically significant. Although there was a significantly less severe overall mood disturbance in the experimental group over time ( P<0.05), this did not apply in the case of anxiety. Such findings suggest that PMRT is a useful adjuvant technique to complement antiemetics for chemotherapy-induced nausea and vomiting and that incorporation of such interventions in the care plan can enhance the standards of care of cancer patients who experience side effects of chemotherapy. PMID- 11904790 TI - The autologous blood and marrow transplant long-term follow-up clinic: a model of care for following and treating survivors of autotransplant. AB - The number of long-term survivors of autologous blood and marrow transplantation (ABMT) is increasing, but little is known about the models of care used to follow these patients. Such information would help in comparison of different methods of follow-up and allow other centers to create programs to follow their survivors. Here we describe a multidisciplinary long-term follow-up clinic for survivors of ABMT and report patient satisfaction with the clinic and economic analysis 1.5 years after its inception. In its first 1.5 years of operation, 83 new patients were seen in the clinic. Patients were evaluated a median of 4.0 years after ABMT (range 4.9 months to 12 years) and were a median of 48.5 years (range 22.5-69.0 years) old. Patients received their ABMT for non-Hodgkin's lymphoma ( n=26), Hodgkin's disease ( n=18), acute myeloid leukemia ( n=18), multiple myeloma ( n=15), breast cancer ( n=5), or desmoplastic round-cell tumor ( n=1). Patient satisfaction was assessed 1.5 years after the establishment of the long-term follow-up clinic by means of a self-report questionnaire. Seventy-five percent of patients returned the survey. Waiting times in the clinic were short, with 70% of patients seen within 30 minutes of arrival. Overall, patients were very satisfied with the new clinic model, with 85% patients reporting that the clinic met their expectations. An economic analysis indicated that the estimated annual cost of operating the clinic was approximately $53,000, which translates into $440 per patient visit, $962 per new patient or $172 per patient visit with the clinic operating at peak capacity. In conclusion, we present an inexpensive model of care for ABMT survivors that can be extended to adult survivors of malignancy not treated by ABMT. PMID- 11904791 TI - Successful treatment of a catheter-related right atrial thrombosis with recombinant tissue plasminogen activator and heparin. AB - Deep venous thrombosis is a possible complication of indwelling central venous catheters (CVC), with an incidence as high as 61%. We report a case of successful thrombolysis of a CVC-related right atrial thrombus in a pediatric cancer patient with recombinant human tissue plasminogen activator (0.1 mg/kg per h for 12 h) and heparin (10 IU/kg per h for 24 h) administered for 6 days. Daily echocardiographic examination showed progressive lysis of the thrombus. The thrombolytic treatment was associated with mild oozing from the venipuncture sites, but no major bleeding was noted; moreover, thrombin, thromboplastin time and fibrinogen were normal or only minimally altered. Anticoagulant therapy, with or without CVC removal, is the treatment of choice for uncomplicated CVC-related thrombosis. Fibrinolytic therapy may be indicated in some cases at risk of pulmonary embolism or to avoid open heart surgery. Recombinant human tissue plasminogen activator is increasingly used for thrombolytic treatment of organ and limb thrombosis, but experience with it in the pediatric hematology-oncology setting is still limited. This report showed that administering recombinant human tissue plasminogen activator in a pediatric cancer patient prior to hematopoietic stem cell transplantation was effective and safe under strict biochemical and instrumental monitoring. Further studies are needed to determine the best antithrombotic treatment for CVC-related thrombosis, and also the dosage of the medication selected and the duration of treatment. PMID- 11904792 TI - Thromboembolic complications related to indwelling central venous catheters in children with oncological/haematological diseases: a retrospective study of 362 catheters. PMID- 11904793 TI - Is there really a need for anti-coagulant prophylaxis for CVC-related thrombotic complications in pediatric oncological-hematological diseases? PMID- 11904796 TI - Extracranial cerebrovascular revascularization for chronic ocular ischemia. AB - We investigated the demographics, presentation, and outcome of patients undergoing cerebrovascular reconstruction for chronic ocular ischemia (COI) at a single institution through a review of 17 patients over a 9-year period. A total of 558 extracranial cerebrovascular reconstructions were performed during the period of study. Seventeen patients (3%) suffered symptoms of COI. There were 19 symptomatic eyes and 15 asymptomatic eyes. Two patients suffered bilateral symptoms. Eighteen (95%) symptomatic eyes experienced rapidly degenerating global visual acuity, and one suffered bright-light amaurosis. Concomitant ocular pathology was present in 10 (59%) patients, consisting of glaucoma (n = 4), cataracts (n = 4), diabetic retinopathy (n = 3), and macular degeneration (n = 1). Symptomatic eyes were found to have significantly worse ipsilateral internal carotid artery (p = 0.004), external carotid artery (p = 0.002), aortic arch branch disease (p = 0.04), and vertebral artery disease (p = 0.04). All 17 reconstructions treated ipsilateral disease. Twelve patients (70.6%) had significant bilateral disease at the time of operation. Three patients underwent staged contralateral reconstruction. Following revascularization, subjective visual improvement or stabilization occurred in 16 patients (94%). A single patient worsened after developing acute narrow angle glaucoma in the perioperative period. Worse cerebrovascular disease is present ipsilateral to symptomatic eyes. When revascularization is performed, arrest of progression or improvement of symptoms occurs in most patients. PMID- 11904798 TI - Carotid artery patch angioplasty: impact and outcome. AB - Our study objective was to determine if patch angioplasty after carotid endarterectomy decreases the incidence of post-reconstruction technical defects and recurrent stenosis. This was a retrospective review of a prospectively maintained database from February 1980 to February 2000. Main outcome measures included incidence of intraoperative technical defects, residual disease within 3 months of endarterectomy, and early/late carotid restenosis >50%. During the study period, 71% (1053) of patients had primary closure and 29% (435) had patch closure. Immediate post-reconstruction intraoperative imaging with angiography or duplex ultrasound was accomplished in all cases. Technical defects prompted the reopening of 136 (13%) carotid arteries closed primarily but only 9 (2%) of those that were patched (p < 0.0001). There were no instances of residual disease in either group. Overall rate of recurrent stenosis was 2%, 3%, and 3.5% at 5, 10, and 15 years, respectively by life-table analysis. Early and late restenosis was significantly reduced by patch angioplasty (p = 0.024 and 0.006, respectively). This study demonstrates that carotid artery patch angioplasty significantly reduces the incidence of detectable technical defects and the early/late recurrent stenosis rate. PMID- 11904797 TI - EEG changes during awake carotid endarterectomy. AB - To determine the reason for differing shunt rates based on electroencephalographic (EEG) and neurologic changes during general and regional anesthetic, respectively, we compared simultaneous EEG tracings and neurologic status in 135 patients undergoing carotid endarterectomy (CEA) under cervical block over a 30-month period. The decision to shunt in these patients was made on the basis of neurologic changes only irrespective of EEG findings. This group was then compared to the 288 patients undergoing CEA under general anesthetic with EEG monitoring over the same period. EEG changes occurred in 7.4% of awake patients and 15.3% of asleep patients (p < 0.03). The rates of ipsilateral hemispheric changes were similar, but no awake patient manifested global EEG changes with clamping while 3.5% of patients under general anesthesia did (p < 0.04). Global, but not hemispheric, changes were correlated with systolic blood pressure variability during clamping. This implies that global EEG changes in anesthetized patients may be the result of the anesthetic technique itself, and that cervical block may in fact be cerebroprotective. PMID- 11904799 TI - Superficial femoral-popliteal vein as a conduit for brachiocephalic arterial reconstructions. AB - Revascularization of brachiocephalic arteries with prosthetic graft offers excellent patency for most reconstructions. For complex brachiocephalic reconstructions, such as redo operations or reconstructions for infection, autogenous conduit may be preferable. Occasionally saphenous vein is inadequate or absent. The purpose of this study was to evaluate the indications and intermediate-term outcomes of superficial femoral-popliteal vein (SFPV) as an alternative conduit for brachiocephalic reconstructions. Over a 6-year period, 71 patients underwent carotid, subclavian, or axillary artery bypass. In 18 (25%) of these reconstruction SFPV was used as the conduit. Ten bypasses (55%) were redo operations. Three bypasses (17%) were performed after failed prosthetic grafts. Three grafts (17%) were required in infected patients. Indications for the use of SFPV included inadequate saphenous vein (n = 13), infection (n = 3), and failed prosthetic bypass (n = 3). Thirty-day mortality was 5.5%. The neurologic event rate was 5.5%. During a mean follow-up of 26 +/- 5 months, there were no graft thromboses or graft infections. Revision-free primary patency was 92% at 48 months. Assisted primary patency was 100%. These data suggest that SFPV is a safe, durable conduit for brachiocephalic reconstructions. SFPV yielded excellent results for a disadvantaged patient population. PMID- 11904800 TI - Endovascular repair of thoracic aortic aneurysms: stent-graft fixation across the aortic arch vessels. AB - The close proximity of the arch vessels to the origin of many thoracic aortic aneurysms (TAA) may result in placement of the stent struts across the left subclavian or carotid ostia. The purpose of this study is to determine the incidence and impact of transaortic arch vessel fixation during thoracic aortic stent graft deployment for the treatment of descending TAA. From May 1997 to July 2000, 20 patients (10 men, 10 women, mean age 82 years) with descending TAA were treated in the operating room with endoluminally placed stent grafts secured proximally to the thoracic aorta with a long (15-mm) uncovered stent segment (Talent LPS). Pre- and post-operative angiograms and IV contrast-enhanced spiral CT scans were performed in all cases. Follow-up contrast CT scans were obtained at 1, 3, 6, and 12 months and yearly thereafter to assess the adequacy of repair and to determine stent position and arch vessel patency. We found that thoracic aortic endograft fixation across the left aortic arch vessels occurs frequently during device placement and is associated with no early morbidity. Long-term follow-up is necessary to ensure that there are no late sequelae. PMID- 11904801 TI - Can the internal iliac artery be safely covered during endovascular repair of abdominal aortic and iliac artery aneurysms? AB - Aneurysmal involvement of the common iliac (CIA) or the internal iliac arteries (IIA) have been relative contraindications for safe endovascular aortic aneurysm (AAA) repair. Our goal was to review our experience in dealing with this problem by performing permanent coverage of one or both IIA during endoluminal repair of aneurysms of the aortoiliac region and to develop a safe, durable strategy. Of the 228 consecutive patients who had endoluminal repair of abdominal aortic (AAA) and iliac artery (IAA) aneurysms between 4/1999 and 4/2001 at our institution, 49 patients underwent coverage and/or coil embolization of one or both IIA during repair because of complex aortoiliac anatomy. These patients were evaluated prospectively for short-term adverse outcome. The results showed that CIA or IIA aneurysms can be managed safely during endoluminal repair of AAA. The IIA can be covered or embolized with minimum adverse consequences in patients who have inadequate CIA for deployment of the aortic or iliac endograft. Unilateral IIA occlusion is well tolerated. We advocate that whenever bilateral IIA occlusion is necessary during endovascular aneurysm repair, one of the IIAs should be revascularized if it is not aneurysmal. PMID- 11904802 TI - Endoleak: predictive value for aneurysm growth at 3 years. AB - Endoleak is a unique radiographic finding after endovascular aneurysm repair. The prognostic implication of endoleak on aneurysm therapy outcome is unknown. Patients with 3 years of follow-up were examined to determine the predictive value of endoleak, as determined by the treating physician, for aneurysm growth. Patients enrolled in a clinical trial for a unibody, bifurcated endovascular graft (Ancure-Guidant/EVT, Menlo Park, CA) were examined with respect to endoleak, as determined by the primary investigator, and aneurysm diameter change. A total of 80 patients were available at 3 years for evaluation. CT scans and ultrasound were used to determine endoleak at discharge, at 6 months, and annually. Patients were categorized as no leak (NL; n = 59), early leak (EL, leak identified by 6 months; n = 15), and late leak (LL, leak identified at 12 months or later; n = 6). A change of 5 mm in transverse diameter relative to the original diameter was used to determine an increase or decrease. Therapeutic intervention for endoleak was analyzed separately in each group. From the results we were able to determine that most patients with distal type 1 or type 2 endoleak have shrinking or stable aneurysms. Endoleak is a poor predictor of aneurysm growth but is statistically associated with enlargement. Absence of endoleak is strongly, but not entirely, predictive of lack of aneurysm growth. Endoleak is a risk factor for aneurysm enlargement, warranting further investigation to examine the etiology of the image, but cannot be used as an endpoint for effective endovascular aneurysm treatment. PMID- 11904803 TI - Percutaneous endovascular abdominal aortic aneurysm repair. AB - In this prospective, nonrandomized study, we compared outcome with percutaneous femoral artery closure to that with open femoral arteriotomy in 95 patients who underwent endovascular AAA repair. Devices were introduced using 22 Fr and/or 16 Fr sheaths. The 8 Fr/10 Fr Perclose devices (Perclose Inc., Redwood City, CA) were used in an off-label "preclose technique." Thirty-three patients had bilateral open femoral arteriotomies, 44 patients had bilateral attempted percutaneous closure, and 18 patients had open femoral arteriotomy on one side and attempted percutaneous closure on the other side. Percutaneous closure was successful in 85% (47/55) of 16 Fr sheaths and 64% (29/45) of 22 Fr sheaths (p < 0.027). Bilateral percutaneous closure was successful in 63% (28/44) of patients. Conversion to open femoral arteriotomy due to bleeding occurred in 24 of 106 percutaneous attempts. There were no dissections, arterial thromboses, or pseudoaneurysms associated with percutaneous arterial closure. Wound complications were seen in 3.6% (3/84) of open arteriotomies and 0.9% (1/106) of all percutaneous attempts and arterial closures (p > 0.05). Gender, previous femoral access, obesity, and iliac occlusive disease were not predictive of percutaneous failure. Procedural success for percutaneous AAA repair is affected by sheath size. Devices delivered through 16 Fr or smaller sheaths will have successful femoral artery closure rates of at least 85%. PMID- 11904804 TI - Are type II (branch vessel) endoleaks really benign? AB - The natural history and clinical significance of type II or branch vessel endoleaks following endovascular aortic aneurysm (AAA) repair remain unclear. Some investigators have suggested that these endoleaks have a benign course and outcome and that they can be safely observed. The purpose of this study was to document the natural history and outcome of all type II endoleaks that have occurred following endovascular AAA repair at our institution. A review of a prospectively compiled database of all endovascular AAA repairs performed at our institution was performed. From this review, we determined that type II endoleaks appear to have a relatively benign course, with a reasonable chance of spontaneously sealing within a 2-year period. No cases of rupture or aneurysm enlargement were documented in patients with open type II leaks. However, almost one-third of the patients did not manifest a type II leak until after their initial CT scan. The implications of such a "delayed" leak are unclear. Careful follow-up remains mandatory in patients with type II endoleaks to better define outcome. PMID- 11904805 TI - Gender does not influence outcomes after iliac angioplasty. AB - The current study was undertaken to evaluate the potential influence of gender on iliac angioplasty outcomes. All iliac angioplasty procedures performed at a tertiary care center from 1994 to 1999 were reviewed. One hundred four angioplasties with or without stenting were performed in 44 women (56 limbs) and 40 men (48 limbs). Age and atherosclerotic risk factors were similar in men and women. Iliac angioplasty was performed for limb salvage in 41% of patients (39% female vs. 44% male; p = 0.65). There were no differences in degree of stenosis, lesion length, or initial angioplasty site. Female iliac arteries were more likely to be occluded (21% vs. 6%; p = 0.03); mean iliac artery luminal diameter was smaller in women than in men (6.5 +/- 0.5 mm vs. 8.2 +/- 0.6 mm; p < 0.001). After a median follow-up of 13 months, there were no significant differences in 2 year primary patency, endovascular primary-assisted patency, or limb salvage rates between women and men. Despite having smaller iliac arteries and a higher incidence of arterial occlusion before treatment, women had outcomes similar to those of men after iliac angioplasty. The current results support the initial use of angioplasty to treat common and external iliac artery occlusive disease in both women and men. PMID- 11904806 TI - Prospective evaluation of endoluminal venous stents in the treatment of the May Thurner syndrome. AB - The May-Thurner syndrome is an acquired stenosis of the left common iliac vein causing pain, edema, or deep venous thrombosis (DVT). The patency and behavior of endoluminal venous stents for this condition was evaluated in this study. Patients with the May-Thurner lesion treated with endoluminal stenting from 1997 to 2000 were evaluated according to an institutional review board-approved protocol. Wallstents (n = 14) or Smart stents (n = 1) were placed into the left common iliac. Patency was evaluated with duplex ultrasonography using a 5 mHz linear array probe (HP 4500) at 6-month intervals. Our results showed that treatment of the May-Thurner syndrome with endoluminal stenting is associated with low morbidity and high patency rates. Longitudinal evaluation of this group of patients is ongoing to confirm these findings. PMID- 11904807 TI - Nitric oxide in experimental aneurysm formation: early events and consequences of nitric oxide inhibition. AB - Mounting evidence suggests that nitric oxide (NO) plays an important role in aneurysm pathogenesis. Nitric oxide synthase (NOS) expression, hemodynamic consequences of NO inhibition, and the effect of NO on matrix metalloproteinase (MMP) expression during aneurysm formation are unknown. In this study, a standard intraaortic elastase infusion rat model was used. Control animals received intraaortic elastase infusion and intraperitoneal saline injections. Experimental groups received intraaortic elastase infusion and intraperitoneal injections of aminoguanidine (500 mg/kg) or L-NAME (2 mg/kg or 10 mg/kg). Aortic diameter, blood pressure, and NO metabolites were measured at surgery and postoperative (POD) 7. A second series of rats were randomly infused with intraaortic elastase or saline and aortas were analyzed on POD 1, 3, and 7 with Western blotting for iNOS, eNOS, and MMP-9 expression. Infusion of elastase produced aneurysms (p > 0.0001) in all rats. Inhibition of NO with aminoguanidine or L-NAME limited aneurysm expansion in all groups (p > 0.05). Nitric oxide metabolites were increased (p < 0.003) in control rats on POD 7. Arterial hypertension was present in all treated animals (p < 0.05). Early up-regulation on POD 1 of iNOS (p < 0.003) was noted in elastas-infused animals, but there was no iNOS expression with saline infusion. MMP-9 expression was present in both groups, with a significant increase in expression for elastase-infused animals noted on POD 7. iNOS expression is up-regulated early in experimental aneurysm formation, followed by increases in MMP-9 expression. Inhibition of NO limits aneurysmal expansion despite production of arterial hypertension. PMID- 11904808 TI - Small-caliber mesothelial cell-layered polytetraflouroethylene vascular grafts in New Zealand white rabbits. AB - Reduction in the thrombogenicity of small-caliber synthetic vascular grafts by lining them with mesothelial cell has been suggested as a method to reduce thrombosis. The purpose of this research is to determine whether creation of a mesothelial lining on the inner surface of a synthetic vascular graft would improve the patency rate of a small-caliber vascular grafts. Carotid interposition grafting was performed using mesothelial-lined grafts (MLG) in 30 New Zealand rabbits and compared with similar carotid interposition grafts using non-mesothelial-lined grafts (NLG) on the contralateral side. The mesothelial lining was created by suturing a piece of harvested peritoneum with the visceral surface toward the lumen onto a 2-mm polytetraflouroethylene (PTFE) graft. Graft patency was studied by in vivo Dopler. In vitro evaluations were done with hematoxylin-eosin stains, broadband cytokeratin staining, and monoclonal antibodies for macrophages. Explanation of the grafts was done in terminal operation at 7, 14, and 21 days. The MLG showed progressive fibroblastic proliferation in direct proportion to the age of the graft, but this did not lead to graft occlusion. However, a significant number of NLG were not patent at each time period studied. We concluded that mesothelial cell lining of smallcaliber PTFE grafts could enhance the short-term patency more than using the PTFE without the mesothelial lining. The use of such hybrid small-caliber grafts has a potential for improving the patency of these artificial vascular graft substitutes. PMID- 11904809 TI - Contrast enhanced duplex ultrasound imaging of the mesenteric arteries. AB - Duplex ultrasound of the visceral arteries is a technically challenging procedure. We examined the clinical usefulness of perflutren intravenous ultrasound contrast to improve the diagnostic accuracy of such studies. Seventeen patients were prospectively studied. A color duplex imaging study of the visceral vasculature was performed with and without the contrast agent. Vessels were imaged and peak systolic velocity and Doppler waveforms of the aorta, celiac artery, superior mesenteric artery, and the inferior mesenteric artery were examined. These results were independently compared to those of contrast angiography. From this analysis we concluded contrast-enhanced duplex imaging of the mesenteric arteries is safe but not routinely required when performed by an experienced sonographer. Ultrasound contrast may be helpful in difficult patients when the vessels are not initially successfully visualized. PMID- 11904810 TI - The fistula elevation procedure: a valuable technique for maximizing arteriovenous fistula utilization. AB - Many patients are not considered candidates for radiocephalic fistula (RCF) or brachiocephalic fistula (BCF) creation or have fistulas that do not mature because the cephalic vein is too deep or tortuous to be accessed. Other patients have not been candidates for the basilic vein transposition (BVT) because limited length of adequate caliber vein precludes subcutaneous tunneling of the vein. In an effort to maximize arteriovenous fistula (AVF) utilization, we developed an adjunctive procedure designed to make the deep or tortuous fistula accessible to needle cannulation. The fistula elevation procedure (FEP) involves mobilization of the fistula, approximation of the subcutaneous tissue beneath the fistula, and subcuticular skin closure over the fistula. The procedure enhances the accessibility of the fistula by placing it in a more superficial position. The overlying cicatrix also acts as a guide for dialysis needle insertion. Between June 1998 and January 2001, 45 patients underwent a natural AVF that could not be accessed secondary to venous depth, tortuosity, or length. In each case, a FEP was performed to salvage the fistula. The FEP was performed as an adjunct to a BCF in 20 patients, a RCF in 7 patients, and a BVT in 8 patients. The FEP is a simple procedure that enhances AVF utilization by making the fistula more accessible to dialysis needle cannulation. The procedure is particularly helpful in obese patients who would not traditionally be considered candidates for natural AVF creation. PMID- 11904811 TI - The transposed forearm loop arteriovenous fistula: a valuable option for primary hemodialysis access in diabetic patients. AB - The distal forearm is the site of first choice for creation of an arteriovenous fistula for hemodialysis. The archetypal procedure, the primary radial-cephalic fistula as described by Brescia, yields excellent functional patency for many patients. Results are much less favorable in patients with diabetes mellitus, for whom non-maturation rates as high as 70% have been reported. This is likely due to inadequate inflow caused by atherosclerotic disease of the forearm arteries in diabetics. Secondary autologous access procedures often involve upper arm configurations such as transposed brachial-basilic fistulas. The present study focuses on a valuable alternative for hemodialysis access in diabetic patients, the transposed forearm loop arteriovenous fistula. Over a 2-year period, 16 forearm loop fistulas were created in 16 diabetic patients who either had a failed radial-cephalic fistula or had arterial anatomy deemed inadequate for wrist fistula formation. In each case, the forearm segment of the basilic or cephalic vein was transposed to form a U-shaped loop and anastomosed to the brachial, proximal radial, or proximal ulnar artery distal to the antecubitai fossa. Functional patency was defined as usability for dialysis. Patency rates were calculated by Kaplan-Meier survival analysis. From our results we determined that the forearm loop fistula is an excellent but underutilized technique that exploits the forearm veins while circumventing the distal arterial supply, thus preserving the upper arm vasculature for future use. PMID- 11904812 TI - Conduit choice for above-knee femoropopliteal bypass grafting in patients with limb-threatening ischemia. AB - Many surgeons consider PTFE to be the conduit of choice for above-knee femoropopliteal bypass grafting, since PTFE is relatively easy to implant and spares autogenous saphenous vein (ASV) for subsequent peripheral or coronary artery bypass grafting (CABG). This practice has recently been challenged, as some studies have suggested that ASV may exhibit superior patency in certain patient subgroups. The purpose of this retrospective study was to examine the contemporary outcome of above-knee femoropopliteal bypass grafting in patients with limb-threatening ischemia. Between January 1995 and December 2000, 159 above knee femoropopliteal bypass grafts were created for limb-threatening ischemia (rest pain or tissue loss). There was a high incidence of comorbid illness, including open foot wounds at the time of operation (62%), hypertension (58%), coronary artery disease (53%), diabetes mellitus (36%), cerebrovascular disease (23%), prior contralateral bypass or amputation (21%), disadvantaged or "blind" outflow (19%), prior ipsilateral bypass (14%), prior CABG (11%) end-stage renal failure (7%). The use of PTFE predominated (n = 11), with a minority of grafts comprising single-segment ipsilateral or contralateral ASV (n = 18). Although the small number of patients undergoing ASV grafting limited the statistical power of comparison, our results suggest that above-knee ASV performs better than PTFE in patients with limb-threatening ischemia. PMID- 11904813 TI - Durability of the dorsalis pedis artery reconstruction in diabetics and nondiabetics: is there a difference? AB - Originally thought to be a disadvantaged outflow source, the dorsalis pedis artery is associated with excellent patency and limb salvage in the diabetic population. The aim of this study was to demonstrate that the dorsalis pedis artery is as durable in nondiabetics as in their diabetic counterparts. During a 21-year period from 1979 to 1999, 299 long lower limb bypasses to the dorsalis pedis artery were performed at our institution. Patient data and outcomes were reviewed from the vascular registry. Statistical analysis and patency rates were compared using chi-squared and log-rank analysis. This study confirms that the dorsalis pedis can be used as an outflow source with durability that is comparable in both diabetics and nondiabetics. No significant difference exists in morbidity, mortality and patency rates. In the nondiabetic, when the dorsalis pedis is the only patent distal vessel, amputation can be avoided if one considers this a suitable option as an outflow source. PMID- 11904814 TI - Lower extremity revascularization without preoperative contrast arteriography: experience with duplex ultrasound arterial mapping in 485 cases. AB - This study reviews our experience with duplex ultrasound arterial mapping (DUAM) for preoperative evaluation in 466 patients (262 men) who underwent 485 lower extremity revascularization procedures from January 1, 1998 to May 30, 2001. Preoperative imaging consisted of DUAM alone in 449 procedures and DUAM and contrast angiography (CA) in 36. An attempt to image from the distal aorta to the pedal arteries was made in all the patients. The selection of optimal inflow and outflow bypasses anastomotic sites was based on a schematic drawing following DUAM examination. Inflow disease was also assessed by intraoperative pressure gradient (IPG) between the distal anastomosis and radial arteries, and completion arteriography of the runoff vessels was obtained, which was correlated with the preoperative findings. Indications for surgery were severe claudication in 91 (19%) limbs, tissue loss in 197 (40%), rest pain in 113 (23%), acute ischemia in 46 (10%), popliteal aneurysm in 18 (4%), superficial femoral artery aneurysm in 1, abdominal aortic aneurysm with claudication in 1, and failing graft in 18 (4%). Age ranged from 30 to 97 years (mean 72 +/- 12 (SD) years) and risk factors such as diabetes, hypertension, use of tobacco, coronary artery disease, and end stage renal disease were present in 45%, 45%, 44%, 44%, and 13% of the patients, respectively. One hundred twenty-one (25%) limbs had at least 1 previous ipsilateral revascularization. The mean DUAM time was 66 +/- 20 (SD) min (30-150 min). Additional preoperative imaging was deemed necessary in 36 cases due to extensive ulcers, edema, severe arterial wall calcification, and very poor runoff. The distal anastomosis was to the popliteal artery in 173 cases and to the tibial and pedal arteries in 255. Inflow procedures to the femoral arteries, embolectomy, thrombectomy, balloon angioplasty, and patch angioplasty accounted for the remaining 57 cases. Overall, 6-, 12-, and -24- month secondary patency rates were 86%, 80%, and 66%, respectively. This early experience shows that high quality arterial ultrasonography performed by a highly skilled vascular technologist may represent an alternative to conventional arteriography for patients in need of lower extremity revascularization. Because of limitations inherent to the technique and very poor runoff observed on ultrasonographic examination, additional preoperative imaging procedure's are needed for certain patients. PMID- 11904815 TI - A decade of decline: an analysis of Medicare reimbursement for vascular surgical procedures. AB - Despite inflation and a robust economy, standard Medicare reimbursements for vascular surgical procedures have progressively declined. The objective of this analysis was to quantitatively and objectively evaluate the decline of vascular surgical reimbursement over the past decade. In this study, data for the analysis of specific vascular surgical procedures was obtained from the National Center for Health Statistics-National Hospital Discharge Survey (NCHS-NHDS) for all vascular procedures as reported by ICD-9-CM codes. The average Medicare reimbursement for each of the specified procedures for 1990 was compared to that of 2001 and the percent change in average reimbursement over this period was calculated. Comparisons between 1990 and 2001 dollar amounts were made after correction for inflation using the consumer price index. This correction factor allows for the calculation of the actual percentage reduction in "real dollars" that is reflected in buying power. We found significant decreases in Medicare reimbursement for each of the vascular procedures included in this analysis. Despite national economic prosperity, there was an average 41% decrease in the buying power per case for vascular surgical procedures over the past decade. We feel that these reductions in reimbursement are overzealous and need to be reexamined. PMID- 11904816 TI - Pelvic lymphocele following motor vehicle collision. AB - The development of lymphocele has been described in the mediastinum following thoracic duct injury from blunt trauma or surgery, in lower extremity surgery or trauma, and in the pelvis following renal transplant or staging lymphadenectomy. We describe a case of pelvic lymphocele following blunt trauma from a motor vehicle collision in which the patient did not sustain any fractures. The patient subsequently experienced right lower extremity pain and swelling thought to result from a deep venous thrombosis. Venogram demonstrated external compression of the right iliac vein, and computed tomography revealed a pelvic fluid collection. The patient underwent successful pigtail catheter placement under ultrasound guidance, and his symptoms resolved completely following 4 weeks of external drainage. A brief review of the diagnosis and management of lymphocele follows. PMID- 11904817 TI - Hyperhomocysteinemia and arterial aneurysm. AB - Hyperhomocysteinemia (HCY) is an independent risk factor for atherosclerosis. Arterial aneurysm has rarely been described in association with heterozygous HCY. Here we report two cases of this association. Case 1 was 32-Year-old man who presented with distal trophic manifestations of the lower extremities. Upon investigation, occlusive arterial disease with fusiform aneurysm of both popliteal arteries and occlusion of the left cubital artery were found. Laboratory findings indicated HCY due to homozygous methylene tetrahydrofolate reductase (MTHFR) deficiency. Case 2 was 38-year-old man with no history of trauma who presented with repeated ischemic events involving the right hand in association with isolated aneurysm of the right cubital artery. Histological study demonstrated extensive dystrophic changes in the aneurysmal vessel wall, including sclerohyalin deposits. The only abnormality was homozygous MTHFR deficiency. Pathologic changes induced by HCY in vessel walls may be implicated in early arterial aneurysm. The association of anatomic lesions, young age, and absence of other causes suggests that the relationship between HCY and arterial aneurysm observed in these two patients was not coincidental. PMID- 11904818 TI - Acute aortic thrombosis following incorrect application of the Heimlich maneuver. AB - The Heimlich maneuver has been widely accepted as a safe and effective method of relieving life-threatening foreign-body upper airway obstruction. When applied incorrectly, however, it may result in direct trauma to the intraabdominal viscera. Only two cases of major aortic complications have been reported. Both have involved thrombosis of an abdominal aortic aneurysm. We report two further instances of aortic thrombotic complications due to the incorrect application of the Heimlich maneuver. The first case resulted in thrombosis of an abdominal aortic aneurysm. In the second case the abdominal thrusts caused dislodgement of thrombus from an atherosclerotic nonaneurysmal aorta, which resulted in thromboembolic occlusion of both lower extremities. In both cases, as with the two previously reported instances, massive reperfusion injury resulted, which eventually proved fatal. When applied incorrectly, the Heimlich maneuver may result in direct trauma to the abdominalaorta and is an unusual cause of acute aortic thrombosis. PMID- 11904819 TI - Adventitial cystic disease of the axillary artery. AB - Adventitial cystic disease (ACD) is an extremely rare cause of arterial and venous insufficiency, with only 317 reported cases in the world literature. These lesions have been previously described in the popliteal fossa, external iliac artery, and distal brachial, radial, and ulnar arteries as well as in the proximal saphenous vein at the ankle. We describe here the first reported case of this disease in a proximal vessel, the axillary artery. A 33-year-old man was evaluated for upper extremity arterial insufficiency and was diagnosed with ACD on the basis of physical examination and radiographic findings, which was confirmed by pathological assessment. The patient was treated by excision of the lesion and interposition vein bypass. As this represents the first case of ACD in the proximal vasculature, it demonstrates that these lesions can occur in axial blood vessels. PMID- 11904847 TI - [The healthy migrant effect: role of selection and late entry bias]. AB - BACKGROUND: First-generation immigrants frequently have a lower mortality than the host population, in spite of a low socio-economic status. This is usually explained by (self-) selection into migration. If this were the case, the immigrants' mortality risk would increase with time under observation. A persistently low mortality could be due to a late entry bias: if migrants are enrolled in a study years after immigration, sick or socio-economically unsuccessful individuals may already have returned to their countries of origin. Mortality risk would then be inversely associated with length of stay in the host country before enrollment. METHODS: We assessed the mortality risk of immigrants from Mediterranean countries to Germany in the German Socio-economic Panel, in relation to time under observation (1-15 years) and length of stay in Germany before enrollment (0-34 years), using the Cox regression. RESULTS: In 1984-98, 2624 immigrants aged 16-83 years accrued 21,858 person years; 59 died. The hazard ratio, adjusted for age, sex and marital status, for each additional year under observation was 0.93 (95 % CI: 0.87-0.99); and for each additional 10 years in Germany before enrollment 0.49 (95 % CI: 0.27-0.89) in the age group >/= 50 years. CONCLUSIONS: We found no evidence for a mortality increase with time under observation, suggesting that the healthy migrant effect is not primarily due to (self-)selection. The initial mortality advantage could be due to international differences in mortality patterns. A late entry bias does contribute to the persisting mortality advantage of older immigrants. PMID- 11904846 TI - [Health in stress: change in the stress concept and its significance for prevention, health and life style]. AB - SUBJECT: The significance of the modern stress concept for health behaviour and prevention. METHODS/RESULTS: Many different definitions of the term "stress" exist concurrently. This observation is important for medicine because of the extensive and increasing use of this term in scientific publications. For a better understanding, the historical development of the stress concept and its association with the biopsychosocial model, with psychoneuroimmunology, mind/body medicine, and prevention/public health is illustrated. "Stress" is interpreted as a more general term that describes the effects of psychosocial and environmental factors on physical or mental well-being. Stressors and stress reactions are distinguished. "Stress" and health are related to health behaviour and life style. CONCLUSIONS: The complex nature of the stress concept requires the actual use of constant definitions. Further, the individual or "subject" needs preventive medical support to strengthen his own capacity and self-care potential so that the balance between stressors and stress reactions, disease- and health promoting factors is achieved. Thus, an integrative, resource- and capacity orientated health care system is implemented. Moreover, letting balance be the target of intervention - and thereby reducing the negative impact of stressors/stress reactions on health - is a possible way of producing a modern, cost-effective, and partnership-like physician-patient relationship. Consequently, "stress" can be "healthy", a challenge, and thereby promote flexibility and transformation. PMID- 11904848 TI - [Does chronic periodontitis play a role in the pathogenesis of cardiovascular and cerebrovascular diseases?]. AB - The role of chronic infections in the initiation of atherosclerotic lesions has been vividly discussed in recent years. A possible causal relationship between cardiovascular diseases and infections with, e. g., Chlamydia pneumoniae, Helicobacter pylori, or herpes viruses had also been established for chronic periodontitis, in particular after discovery of DNA of typical periodontal pathogens in atheromatous plaques. Especially in longitudinal epidemiologic studies, a low or moderate association between existing periodontitis and the development of, e. g., coronary heart disease or non-haemorrhagic stroke had been observed. In this article the respective literature is critically reviewed. In particular, the influence of incomplete or inappropriate adjustment for common risk factors for both diseases, i. e., cardiovascular disease and periodontitis should be analysed. In metaanalyses of prospective studies, in which the respective endpoint occurred after the investigation had commenced, relative risks of periodontitis of 1.12 (95 % confidence interval 0.95-1.33) for coronary heart disease and 1.73 (0.89-3.34) for ischaemic stroke were calculated. Whether chronic periodontitis actually represents an important risk for the development of cardiovascular diseases remains questionable. Already planned intervention studies appear to be premature and ethically highly problematic. PMID- 11904849 TI - [Biological markers in epidemiology: concepts, applications, perspectives (part I)]. AB - The inclusion of biomarkers in epidemiological research provides new possibilities for exposure assessment and the study of early structural or functional changes and pre-clinical stages of diseases. At the same time issues of validity, reliability, and quality control as well as logistics require special attention. Usually epidemiological studies become more expensive with regard to time and cost. Interdisciplinary collaboration between epidemiology, basic research, and laboratory research is crucial. A prerequisite for this collaboration are agreements on definitions, methods and procedures. The definition of "biomarker" and a description of previous uses of biomarkers in epidemiological studies are presented in the first part of this paper. The second part addresses genetic markers and markers of individual sensitivity and susceptibility. We will end with a discussion about the possible future of biomarkers in epidemiology. PMID- 11904850 TI - [Sociopsychiatric care as a municipal problem]. AB - Because of their disease mentally ill persons are often unable to request and receive necessary help on their own. Hence, local authorities have the legal duty to offer support in subsidiary or complementary ways or to organize help together with other participants concerned with psychiatric care. If the transition from community care to the differently structured regular health care system is too fast there is a danger of worsening of the symptoms. In the city of Bochum health insurance bodies agreed to take over the costs of therapy for mentally ill people via the sociopsychiatric services within the scope of crisis intervention. Background information on the sociopsychiatric services of the city of Bochum is given. PMID- 11904851 TI - [Accidents of toddlers and youngsters]. AB - The Public Health Department in Biberach an der Riss developed a questionnaire to investigate the incidence of accidents in children under school-starting age (6 years). This questionnaire was presented to the parents of more than 2,300 prospective first-graders from the town and rural district on the occasion of the pre-school medical examination 2000. As this examination is mandatory for all children starting school, and as the questions were answered by all the parents with very few exceptions (language reasons), a complete survey can be assumed. The investigation confirmed the results of last year: The incidence of children who suffered an accident requiring medical attention before reaching school age is approximately 33 %; boys are predominantly involved. The scene of accidents also changes with increasing age from living quarters to outside areas. The most frequent type of accidents are, of course, falls, resulting especially in injuries to the head and face. Scalds and burns, in particular at the age of 2, occur more frequently in the Biberach district than described in other up-to-date investigations in Germany. For this reason efforts have to be made to reduce this number over the next years. About 11 % of accidents occur in the streets or involve traffic, a result which is also higher in comparison to other investigations. According to the statement of parents, more than two-thirds of accidents are caused by the children themselves, including babies and toddlers. At the time of the accident 40 % of the children were without parental control, and 20 % completely alone.A great number of the accidents could certainly have been prevented. That is why the results of the study should be made available to all those responsible for the care and wellbeing of this age group. The last section of the paper deals with the most urgent needs of action to be implemented in the long run for the sake of the health of our children. PMID- 11904852 TI - [Teachers' health - a challenge for an interdisciplinary prevention concept]. AB - The early retirement of teachers because of illness is presently a great social and sociomedical problem. In Bavaria, currently more than every second case of early retirement in the profession is the result of psychic or psychosomatic illness. The aim of all preventive measures must therefore be first of all the preservation or restoration of psychic health of teachers. With an underlying comprehensive biopsychosocial understanding of health and performance, preventive measures should take into consideration both circumstances and behaviour, as well as pathogenetic and salutogenetic aspects of the disease. The preventive concept developed under these premises is characterised by the terms interdisciplinary, multidimensional, integrative and institutionalised. Interdisciplinary means the concerted effort of all disciplines involved in the health of teachers. Multidimensional implies action at all levels of disease prevention and includes therapy and rehabilitation. Integrative stands for the formation of networks for optimising processes and the linking of interfaces in the sense of case/disability management. Institutionalised means the creation of structures for co-ordinating and controlling. In addition, potential areas of activity for those involved in prevention, in particular also occupational medicine, are described, resulting in an appeal that the insights gained may be translated into practical reality. PMID- 11904853 TI - [Prevalence estimation of overweight and obesity based on subjective data of body mass-index (BMI)]. AB - OBJECTIVES AND METHODS: In connection with the increasing importance of chronic diseases the estimation of the prevalence of overweight and obesity becomes more and more important. Today these estimations are usually done via the Body-Mass Index (BMI). For economic reasons BMI is often obtained by means of questionnaires or interviews. These (subjective) BMI-data show great differences to measured (objective) data. The differences between subjective and objective data and their dependence on age, gender and residence were investigated. RESULTS: Subjective and objective data show significant differences. On the basis of subjective data too many persons classify themselves as underweight or normal weighted and fewer persons classify themselves as overweight and obese. Variance analysis shows significant influences of gender and age. Women underestimate their BMI more than men. With increasing age the differences also increase. CONCLUSIONS: The estimation of BMI based on subjective data is inaccurate. In this way the prevalence of obesity and overweight are underestimated. That is why subjective data are not useful for clinical and epidemiological research, but it is interesting against the background of health psychology. PMID- 11904854 TI - [Problems in defining obesity in prepubescent children: consequences for assessing the requirements for medical rehabilitation]. AB - INTRODUCTION: Prevalence of obesity has been on the increase in recent years. In contrast to obesity in adulthood, childhood obesity is still not uniformly defined. This results in problems to determine the need for medical rehabilitation. METHODS: The Kiel Obesity Prevention Study (KOPS) is a cross sectional study assessing the nutritional status of 5-7 year-old German children. Three different definitions of overweight and obesity (German reference data for triceps skinfold thickness [90(th) percentile] and BMI [90(th)/97(th) percentile] as well as an international standard for BMI [extrapolated to levels of adults]) were applied to 1,643 children of KOPS enrolled between 1996 and 2000 (19 % of all first-graders in Kiel in this period). RESULTS: The prevalence of overweight varies from 9 to 21 % depending on the applied definition. With the definitions of overweight and obesity based on newer BMI percentiles a part of overweight children are not classified as such. The present state of art is that there is only a need for obese children for medical rehabilitation: these are 3.3 and 3.5 % of 5-7 year-old children in Kiel, respectively. CONCLUSIONS: Experts should work out an agreement concerning a uniform definition of childhood obesity. Currently, medical rehabilitation services are offered only to extremely obese children. There is a need for more and earlier preventive measures. PMID- 11904856 TI - [Relationship between subjectively perceived working conditions and alcohol consumption: results of the TACOS study]. AB - The purpose of this investigation was to examine the relationship between subjectively perceived working conditions and alcohol consumption based on a stress coping model. The investigation is part of the study 'Transitions in Alcohol Consumption and Smoking' (TACOS), a representative general population survey in a Northern German region. The current evaluation includes 2,471 working individuals, aged 18-64 years. Data with regard to alcohol consumption were collected via the 'Munich Composite International Diagnostic Interview' (M-CIDI). The three scales of the 'Normative and Subjective Assessment of working analysis' (NUSA) Contents of work, Physical work conditions and Intensity of work gathered differentiated information on subjectively perceived working conditions. The ANOVA results show that alcohol consumption does not account for variance in the scales of subjective working conditions. Further correlation and regression analyses do not reveal a relationship between the scales of NUSA and the quantity of drinking.A substantial relationship between subjectively perceived working conditions and moderate alcohol consumption, alcohol abuse or alcohol dependence could not be established. The reported results appear to indicate complex and multifactorial associations between subjectively perceived working conditions and alcohol consumption. Implications for alcohol prevention in worksite environments are discussed. PMID- 11904855 TI - [Biological Markers in Epidemiology: Concepts, applications, Perspectives (Part II)]. AB - The first part of this paper outlined terms and definitions in the context of the application of biological markers in epidemiological studies. Cardiovascular epidemiology served as an example for their historical development. The second part focusses on DNA-based biomarkers, practical and methodological dimensions of the use of biomarkers in analytic epidemiological studies as well as requirements in respect of validity and quality assurance. Most genetic polymorphisms have no impact on health. However, some can be used as biomarkers for individual sensitvity to exposures and susceptibility for specific diseases. The Human Genome Project has brought about a quantum leap in the development of genetic markers. The practical implications cannot presently be assessed with certainty. However, present and future research programmes of gene-environment interactions depend on "traditional" epidemiological study designs, methods, and concepts. Ethical principles and data protection requirements apply equally to genetic and molecular epidemiology as do the "Guidelines for Good Epidemiological Practice". PMID- 11904857 TI - [Demand for environmental medical advice at public health offices: experiences in the district aachen public health office]. AB - Since November, 1999 environmental medical advice is offered to interested citizens in the Aachen district at the District Aachen Public Health Office in cooperation with the outpatient unit of environmental medicine (UEM) of the Institute of Hygiene and Environmental Medicine of the University Hospital at Aachen, Germany. Advisory cases are documented in a data bank of Microsoft(R) Access 97. Until now, all advisory cases between November, 1999 and March, 2001 have been descriptively analysed. In this period, 34 personal and two telephonic advices were performed. The frequency of advisory activities is in the lower rang of published experiences in environmental medicine. Age distribution, more frequent advice utilization by women than by men and predominance of unspecific health disorders are comparable with published environmental medical experiences. However, in respect of suspected exposures, unspecific indoor-related environmental factors are predominant. In the past this was true for wood preservatives. Judgement about possible relationships between suspected environmental factors and health disorders or diseases was positive among 11.8 % of the persons seeking advice. This percentage is higher than published experiences which mostly show values below 10 %. It must be considered that this judgement depends primarily on the physician. Other reasons may be the too small number of advice seeking persons and selective influences. Furthermore, a definite judgement can be made only after environmental medical diagnostics (biological monitoring, local inspection, ambient monitoring) and differential diagnostics. Conspicuously, 76.5 % of the advisory cases had no contact to environmental medicine prior to the environmental medical advice at the Aachen District Public Health Office. This points to an information deficit about possibilities to clarify questions concerning environmental medicine in the population. In this context a regional guide on environmental medicine may be helpful. The environmental medical advice for citizens is an excellent example of a successful cooperation between a public health office and an university, which have different special experience in environmental hygiene and environmental medicine. This cooperation brings selectively citizens seeking for advice in environment-related health risks and disorders to practitioners specialised in environmental medicine. PMID- 11904859 TI - [Free choice of doctors in Germany in retrospect]. AB - Due to discussions on the cost and quality of health care and a new legislation on the German statutory sickness insurance system in 1999, the free choice of doctors has recently become topical. To assess its legitimation for the German health care system, its history and the groups of interest involved should be taken into consideration. Before the period of industrialization no homogeneous pattern of the medical profession existed. In case of illness individuals who lived within reach and were known for their competence in disease matters were approached. However, industrialization destroyed existing social networks, and establishment of new structures of health care in rural as well as metropolitan areas became necessary. The government approached this challenge by structuring medical education, passing regulations on the settlement of doctors and promoting the foundation of statutory sickness funds. The Health Insurance Law of 1883 established a mandatory insurance system for a broad array of industries. As it was the sickness funds' responsibility to provide sufficient resources for medical care, a sick member was tied to the physician under contract with his insurance. After a rapid increase in practising physicians at the end of the 19(th) century, doctors' organisations were eager to gain access to the new market segment of insurance members by calling for the free choice of physicians. The Leipzig association (Hartmannbund) was founded in 1900 to organize strikes of doctors in order to get their goals accepted. After 30 years of conflicts an appeasement was achieved by a presidential emergency law in 1931. It transferred the responsibility for the provision of sufficient health care resources from the sickness funds to the newly created body of the Association of Sickness Fund Physicians (Kassenarztliche Vereinigung) and determined the patients' free choice among licensed sickness fund physicians. PMID- 11904858 TI - [Monitoring of naevus density--an approximate marker of previous UV exposure--in children at school enrollment as a method to detect shifts in melanoma risk in the population]. AB - OBJECTIVE: Naevus density in children depends both on constitutional factors (see below) and on previous (solar) UV exposure, which is a well-known risk factor for malignant melanoma. Secular trends of childhood UV exposure - e. g., due to intervention programmes - can be monitored by repeated, standardised cross sectional studies assessing naevus density in children. METHODS: The 'CMONDE Study (childhood monitoring of naevus density)' appears as a suitable instrument, introduced into the process of school enrollment for children of the years 1999 and 2000 in Gottingen. RESULTS: Analyses are based on 3883 children. Median age was 6.25 years, the proportion of girls 47 %. Median naevus density was 5.8/m(2), with an increase of density from 'Fitzpatrick skin type' IV to II, but marked decrease in type I. Similarly, naevus density increased with increasing lightness of hair colour, but was very low in red-haired individuals. While the number of freckles was also strongly associated with naevus density, the association between iris colour and skin darkness, respectively, was weak. CONCLUSION: To meet the objective of continual monitoring, further comparative cross-sectional studies are needed. PMID- 11904860 TI - Imigran: Ten years of improving the lives of migraine patients. Rome, 24 March 2000. Proceedings of a conference. PMID- 11904861 TI - Fairness: the most basic outcome measure. PMID- 11904862 TI - Malnutrition: role of the TwoCal HN Med Pass program. AB - Malnutrition is common in older adults and is associated with poor outcomes. The causes and outcomes of malnutrition are discussed, and the TwoCal HN Med Pass program, designed to overcome poor dietary intake, is described. Benefits of the program, role of the pharmacist, identification of candidates for the TwoCal HN Med Pass program, and health care team roles and responsibilities are reviewed. PMID- 11904863 TI - The nurse's role in smoking cessation. AB - Smoking cessation is an important component to improving patients' health and quality of life. This case study demonstrates how nurses can implement a successful smoking cessation program in outpatient settings. By applying this knowledge, adult-health nurses can increase the probability that hospitalized patients will be successful in stopping smoking. PMID- 11904864 TI - Frequency and effectiveness of self-care actions and menopause symptoms of middle aged working women. AB - A sample of working women (462) was surveyed to determine the frequency and effectiveness of menopause-specific symptoms and self-care actions. Perimenopause women were more symptomatic than the pre or postmenopause women. The most frequently used and effective self-care actions were to control vasomotor symptoms. PMID- 11904865 TI - A comprehensive interactive competency program. Part II: Implementation, outcomes, and followup. AB - The department of medicine/oncology nursing of an academic medical center developed and implemented a comprehensive interactive competency program. The program proved to be valuable to the individual nurse and the organization. Part II of this two-part series describes the implementation, outcomes, and followup of the program. PMID- 11904866 TI - Calciphylaxis in chronic renal failure. PMID- 11904867 TI - The new NSAIDs: cox-2 inhibitors. PMID- 11904868 TI - A novel program to assess and manage pain. PMID- 11904870 TI - Pressures on nursing education. PMID- 11904871 TI - Nurse-patient relationships. Essential skills for expert nursing practice. PMID- 11904872 TI - Nursing's socialization of nurses. PMID- 11904874 TI - Socializing students on the complexity of practice. Interview by Carol Lindeman. PMID- 11904873 TI - Self-development and the arts. Interviewed by Carol Lindeman. PMID- 11904875 TI - Nursing's timeless value: the dignity and value of all persons. PMID- 11904876 TI - An ideal education: health promotion and dis-ease prevention. PMID- 11904877 TI - A perspective for 2000: today's realities, tomorrow's needs. PMID- 11904878 TI - The timeless values of nursing. PMID- 11904879 TI - Nursing values: a look back, a view forward. Interview by Marie Manthey. PMID- 11904880 TI - Nursing's timeless value: intentional caring. PMID- 11904881 TI - Nursing's timeless value: advocacy. PMID- 11904882 TI - Nursing's timeless value: promoting community health. PMID- 11904883 TI - Elderly patients: people not 'bed-blockers'. PMID- 11904884 TI - Good practice guidance for continence services. PMID- 11904886 TI - Case 26: tying a client to a toilet. Client with learning disabilities who died after being tied to a toilet. PMID- 11904887 TI - Malnutrition in critically ill patients in intensive care units. AB - The provision of artificial nutrition for critically ill patients is of great importance as many are unable to maintain their own nutritional needs. The administration of total parenteral nutrition (TPN) and enteral nutrition (EN) has become a daily practice in intensive care units. Despite this, many patients remain undernourished or even malnourished and it is estimated that the incidence of malnutrition in intensive care patients could be as high as 50% (McCain, 1993). The reasons by which patients become or remain undernourished are multifactorial and range from physiological to iatrogenic. In order to lessen the catabolic state which results from the hypermetabolism associated with critical illness, prompt and adequate nutritional support must be delivered. It is essential that members of the multidisciplinary team caring for critically ill patients are aware of the importance of nutrition and the deleterious effects of malnutrition to achieve the best possible outcome for patients. PMID- 11904888 TI - Your child has diabetes: hospital or home at diagnosis? AB - Historically, children with diabetes have been hospitalized at diagnosis, but increasingly, newly diagnosed children are being cared for entirely at home. The management of this chronic condition usually involves the whole family, with children often taking responsibility for much of their own care. However, this article focuses specifically on the needs of parents, forming part of an extensive literature review informing a study exploring parents' experience of home management and coping over the first year with childhood diabetes. A search of the literature revealed a scarcity of evidence overall about hospitalization or home management from a parental perspective, and none in relation to childhood diabetes. This article provides a critical appraisal of the appropriateness of these two approaches to care for parents of children with newly diagnosed diabetes. First, a brief introduction to home management in childhood diabetes is followed by an examination of the small amount of research found about home management and hospitalization from the point of view of parents. Then, the possible benefits and disadvantages of both approaches are discussed and subsequently scrutinized in the context of childhood diabetes. Finally, preliminary conclusions are drawn and suggestions made for the direction of future research in this area. PMID- 11904889 TI - Accountability: a fundamental component of nursing practice. AB - The nursing care of the patient makes great demands on nurses' knowledge and skills; the care given must be competent, efficient and effective. In order to achieve this, nurses must have the necessary authority to manage and control their practice. This article seeks to examine the concept of accountability and relates its relevance to everyday nursing practice. A theoretical and critical incident approach is employed to demonstrate the principles surrounding accountability within nursing. PMID- 11904890 TI - Male patients with lower urinary tract symptoms. 2: Treatment. AB - The first part of this article (Dorey, 2000) described the subjective and objective assessment of men with lower urinary tract symptoms (LUTS). This article will examine treatment protocols for stress incontinence, urge incontinence, post-prostatectomy incontinence, post-micturition dribble, overflow incontinence, reflex incontinence and functional incontinence. Pelvic floor muscle exercises, biofeedback, electrical stimulation, urge suppression techniques, and fluid intake are discussed. It is concluded that men with LUTS can benefit from conservative treatment. PMID- 11904891 TI - Ageism and the abuse of older people in health and social care. AB - Ageing is a natural process and yet ageism, ageist practice and abuse of older people occur among not only the general public but also in health and social care settings. Recent media reports have highlighted delays in meeting the needs of older people, physical and psychological abuse and that decisions are being made about whether or not to resuscitate an older patient without consultation with the patient and his/her family (e.g. British Journal of Nursing, 2000). This article looks at ageism and the abuse of older people and discusses what can be done to achieve quality care for older people while dealing with obstacles such as poor collaboration between agencies, a lack of support for carers and the belief that the needs of older people are less important than those of the young. PMID- 11904893 TI - Living with a young person who wets the bed: the families' experience. AB - After allergic disorders, bed-wetting is the most common chronic condition of childhood. It can seriously diminish the quality of life of young people and their families, having an impact on day-to-day activities, family holidays and the young person's willingness and ability to stay away from home with friends and wider family. In this ethnographic study, family members describe the practical and social consequences of bed-wetting, both for themselves and for the family, and the methods that they have employed to encourage the bed-wetting to stop. Most of these methods have little chance of success. Many families' feelings of helplessness and isolation are reinforced by lack of help from healthcare professionals, although the professional's intention to be helpful is rarely questioned. The nature of the families' experiences illustrates the urgent need for adopting a new professional approach to the support of these families, which is based on the principles of 'family nursing'. PMID- 11904892 TI - Autonomy and clinical practice. 2: Patient privacy and nursing practice. AB - This article, the second in a series of three considering issues of autonomy, privacy and informed consent in nurse/patient interactions, focuses on the wider conceptions of patient privacy and confidentiality. Given that patients in institutional care are likely to suffer intrusions into their privacy which would be considered unusual in normal social interaction, it is interesting to note the dearth of literature in this area. Some definitions of privacy are considered in an attempt to begin to raise readers' awareness of the complexity of this notion. It can be argued that privacy is a pertinent notion to consider, both in order to gain a greater understanding of what is meant by the term and in terms of the implications of this understanding for clinical practice. PMID- 11904895 TI - UKCC will now explain its misconduct decisions. PMID- 11904896 TI - The reflective leader. PMID- 11904894 TI - Pathways for continence care: background and audit. AB - This article is the first in a series of three covering the use of care pathways for continence care. Trusts in Basingstoke, Swindon and Salisbury have collaborated in supporting their continence advisers in moving from finance driven assessment data to evidence-based care pathways and the provision of patient information. This article identifies the background and approach to care pathways and addresses the quality issues. It details the issues facing continence advisers and how care pathways may help to address them. Furthermore, it describes a baseline audit which was carried out to ensure that facts rather than beliefs were being used and this demonstrated that little advice or treatment was actually reaching the patient. PMID- 11904897 TI - Reflective practice and the Arizona Nurses Association. PMID- 11904898 TI - Julie. A preceptor reflects on one who taught her. PMID- 11904899 TI - Reflective practice. PMID- 11904900 TI - Reflective practice in the UK. Interview by Marie Manthey. PMID- 11904901 TI - Reflective practice for American nurses. Interview by Mae McWeeny. PMID- 11904902 TI - End-of-life care: the nursing role. PMID- 11904903 TI - Artificial nutrition and hydration at the end of life. AB - Patients who are terminally ill do not respond to administration of artificial nutrition and hydration in the same way as patients who have potential for recovery. Knowledge of end-stage disease and ethical and legal parameters are necessary for health care team members to make clinically and morally sound therapeutic decisions in conjunction with the patient and family. PMID- 11904904 TI - Esophageal cancer: an overview. AB - Esophageal cancer is the second most common solid intrathoracic malignancy behind lung cancer. Treatment of esophageal cancer is dependent upon the stage of presentation and the options include chemotherapy, radiation therapy, surgery, multimodality therapy, or palliative care. Early detection of this disease is the primary method to decrease its high morbidity and mortality. PMID- 11904905 TI - Primary pulmonary hypertension: a review for advanced practice nurses. AB - Primary pulmonary hypertension (PPH) is a rare entity of unknown etiology, manifested as narrowing and obstruction of the pulmonary arteries leading to elevated pulmonary artery pressures. Left untreated, these high pressures can lead to right ventricular dilation, right heart failure, and death within 3 to 5 years. Over the last decade, advances have been made in the diagnosis and treatment of PPH. Advanced practice nurses and other expert nurses can use and interpret diagnostic and therapeutic medical regimens currently available to identify and treat this rare condition. PMID- 11904906 TI - A comprehensive interactive competency program. Part I: Development and framework. AB - The department of medicine/oncology nursing of an academic medical center developed and implemented a comprehensive interactive competency program. The program proved to be valuable to nurses and the organization. Part I of this two part series describes the process of program development and the resulting framework. PMID- 11904907 TI - Withholding and withdrawing artificial nutrition and hydration--a legal perspective. PMID- 11904908 TI - Evidence-based medicine needs to be promoted more vigorously. PMID- 11904909 TI - Plans, providers should use HIPAA breathing room wisely. PMID- 11904910 TI - Health care cost control. Getting on the right track. PMID- 11904911 TI - Proposal to regulate formularies draws sharp difference of opinion. PMID- 11904912 TI - Push is on for improved treatment of women's psychological maladies. PMID- 11904913 TI - Reframing the pharmaceutical manufacturer/health plan relationship in managed care. AB - Managed care is stuck in a vendor stage of health care industry evolution that is organized, primarily, to beat back costs through contracted discounts and utilization management. At the same time, the potential exists for an altogether different managed care that is based on a more explicit mission of lowering costs through improved quality. The foundation for this alternative approach is evident in current practices involving disease management and clinical-quality improvement. Significantly, while health plans do not have the staff or capital to systematically adopt these practices, pharmaceutical companies are in a unique position to assist. To the degree health plans start building sophisticated, long term strategic partnerships with pharmaceutical manufacturers into their business models, managed care will gain the capacity to advance beyond its vendor stage and make good on its original mission of promoting preventive health, improving individual outcomes, and realizing sustainable cost containment. For this to happen, it is suggested that health plans address "total cost-of-care savings" in their budget process, and the pharmaceutical manufacturers establish a "consultative service function" in their managed care divisions. PMID- 11904914 TI - Why you should care about improving clinical practice. PMID- 11904915 TI - E-prescribing gets more enticing. PMID- 11904916 TI - Laws governing peer immunity, physician credentialing upheld. PMID- 11904917 TI - Managed care outlook. Where HMOs go head to head. PMID- 11904918 TI - [Immunochemical characterization of Fraction I of Yersinia pestis strains by CAF1M gene]. AB - The role of caf1M gene in biogenesis of Yersinia pestis capsule was studied in natural strains of the agent with Fra+/- phenotypes and recombinant variants with ycaA (caf1+;caf1M;caf1A+;caf1R+) locus defect. These bacteria did not form a clearly discernible capsule stained by classical methods but synthesized Cafl, whose content in the cells was many times higher than in lysates, in external cell wall, and in the medium with reference Y. pestis EV NIIEG culture (caf1+;caf1M;caf1A+;caf1R+). However Caf1 was not detected on the surface or culture fluid of natural and mutant Y. pestis cells. Exclusive role of Caf1M in Caf1 delivery to Y. pestis cell surface, but not in F1 monomer folding, was proven. Retention of lipopolysaccharide (LPS), a typical SR-LPS configuration and epitope specificity of its components was demonstrated, ensuring similar reactivity in solid-phase enzyme immunoassay with a panel of monoclonal antibodies to Y. pestis LPS. Study of immunochemical properties of antigenic substances derived from caf1M-defective Y. pestis cells by isolation of F1 showed that these substances represent an envelope protein involved in the caf1+ strains (together with Caf1) in assembly of "mature" F1 molecule as a result of posttranslation modification of various genes products. Variants of identification of Y. pestis with Fra+ phenotype by means of monoclonal antibodies to F1, fibrinolysis/coagulase, or LPS in solid-phase enzyme immunoassay are discussed. PMID- 11904919 TI - [Molecular construction of experimental combined vaccines]. AB - A method for designing molecular constructions of combined artificial immunogens mimicking viral particles is proposed. Using this method, it is possible to expose antigenic determinants of any viral protein on the surface of such particles and to deliver plasmids containing genes encoding the synthesis of protein antigens to target cells. This approach was used to create the constructions of combined artificial immunogens inducing the production of specific antibodies to HIV-1 and to evaluate the extent and duration of B cellular immune response depending on the way of antigen exposure to immunocompetent cells. PMID- 11904920 TI - [Estimation of the efficiency of ERIC-PCR typing of Mycobacterium tuberculosis for evaluation of the prevalence of antibiotic-resistant strains]. AB - Twenty-four antibiotic-resistant and sensitive strains of M. tuberculosis isolated from different territories of the Irkutsk region (East Siberia) were studied using PCR genotyping by enterobacterial repetitive intergeneric consensus (ERIC). Evolution relationships are illustrated by phylogenetic trees as a result of analysis by UPGMA and ML approaches. It was found that the studied samples belonged to two genetically different groups, both of which included sensitive and resistant strains. The sensitivity of the method was calculated by the Hunter Gaston index. Based on these data, a probable pattern of emergence and propagation of antibiotic-resistant forms of tuberculosis in the studied region is discussed. PMID- 11904921 TI - [Mutation and recombination variability of vaccine polioviruses isolated in Belarus (1960-1999)]. AB - One hundred and eight vaccine-derived strains of types 1, 2, and 3 poliovirus (25 PV1, 34 PV2, and 48 PV3) isolated in Belarus in 1960-1999 were analyzed by double restriction fragment length polymorphism assay (RFLP-1, -3D1). Forty-four (40.7%) of strains were genetically modified. Eight (7.4%) PV were modified by mutation, 16 (14.8%) by recombination, and 20 (18.5%) by both mutation and recombination. The genomes of 16 PV were analyzed by multiple RFLP technique covering VP1-, VP1/VP2A-, P2-, 3AC-, and 3D1-coding regions. The majority of recombinants were "simple" (with one crossing over site). One strain was "double" recombinant (two crossings over sites) and one more "multiple" recombinant (three crossing over sites). Partial nucleotide sequencing of some recombinant strains showed that the degree of these strains' divergence was less than 1% in comparison with the original vaccine viruses. PMID- 11904922 TI - [Modern concepts on the relationship between the agents causing plague and pseudotuberculosis]. AB - The authors present published data and their own findings on the relationship between Yersinia pestis and Y. pseudotuberculosis and on the origination of Y. pestis from Y. pseudotuberculosis. Study of microbiological and biochemical characteristics, external membrane protein spectra, and stability of chromosomal region of pigmentation brought the authors to a hypothesis that Y. pestis minor subspecies (ssp. caucasica, altaica, hissarica, ulegeica) which are characterized by selective virulence occupy an intermediate position between Y. pseudotuberculosis and basic species of Y. pestis. PMID- 11904923 TI - [Possibility of using fusion proteins for detection of nonsense mutations and frame-shift mutations in BRCA1 gene]. AB - The aim of this study was to evaluate the possibility of detecting nonsense and frame-shift mutations in exon 11 of brca1 gene by constructing fusion open reading frame (ORF) "exon 11 ORF-alpha-peptide of beta-galactosidase". The ability/inability of this newly constructed ORF to cause alpha-complementation in E. coli delta M15gal cells transformed by the plasmid with the ORF may reflect the absence/presence of nonsense and frame-shift mutations in the studied fragment. A single ORF fragment of exon 11 of brca1 gene--LacZ' gene was designed in pGEN7Zf plasmid, the plasmid was shown to cause Lac+ phenotype in E. coli delta M15gal. Four frame-shift deletion mutations were introduced into exon 11 sequence in the plasmid. Surprisingly, the frame-shift deletion mutations did not influence the ability of plasmids to induce Lac+ phenotype in E. coli delta M15gal in 3 cases and only one deletion mutation resulted in inability of the plasmid to form Lac+ phenotype in E. coli delta M15gal. We suppose that the phenomenon can be explained by the alpha-peptide translation reinitiation from inframe ATG codons situated within the exon 11 sequence. Seven inframe ATG sequences were found in exon 11, at least two in-frame ATG-containing fragments were demonstrated to cause reinitiation. On the other hand, the only deletion mutation resulted in inability of the plasmid to form Lac+ phenotype in E. coli delta M15gal did not leave LacZ' in-frame ATG in econ 11 sequence. We conclude that it is possible to detect frame-shift mutations by in-frame cloning with the LacZ' reporter gene, but this possibility is strongly impeded by the reinitiation of alpha-peptide translation from the in-frame ATG codons within the studied sequence. PMID- 11904924 TI - [Burkholderia thailandensis: biological properties, identification and taxonomy]. AB - Burkholderia pseudomallei-like microorganisms have been isolated from soil and water in regions with endemic melioidosis. These strains have biochemical and antigenic profiles identical to melioidosis agents, except that they differ by virulence and L-arabinose (vir-, ara+). There are minor differences between these species by rRNA sequence. DNA hybridization and, more so, positive transformation of DNA auxotrophic mutants of B. pseudomallei by cell lysates of B. thailandensis and B. mallei confirmed the homology of these species' genomes. These members of the Burkholderia genus (pseudomallei, mallei, and thailandensis) can be regarded as a supraspecies taxon: pseudomallei group. B. thailandensis strains are not virulent for guinea pigs and slightly virulent for golden hamsters. Immunization with live cultures of B. thailandensis protected more than 50% guinea pigs challenged with 200 LD50 B. pseudomallei 100. B. thailandensis is suggested as a potential melioidosis vaccine. PMID- 11904925 TI - Nursing case management: quality and cost. PMID- 11904926 TI - Nurses and personal journal writing. PMID- 11904927 TI - A speedy return to familiar quarters. PMID- 11904929 TI - New leadership work: translating the work of staff nurses into the marketplace. PMID- 11904930 TI - Nursing and partnership economics. Interview by Kathy Malloch. PMID- 11904931 TI - An investment in labor. PMID- 11904932 TI - Try a good book bibliotherapy as spiritual care. PMID- 11904933 TI - Sherry's story. Facing MS faith & cognitive restructuring. PMID- 11904934 TI - God's special children. PMID- 11904935 TI - "Why don't you treat me any different?" Caring without partiality. PMID- 11904936 TI - Suffering dialogues with five faiths. PMID- 11904938 TI - Bringing death back home. PMID- 11904937 TI - Were you there? Healing in the cross of Christ. PMID- 11904939 TI - Healing hands a celebration. PMID- 11904940 TI - Losing more than a limb. How can we help? PMID- 11904941 TI - Scratching where it itches. PMID- 11904942 TI - Professionalism and ministry: can we keep our balance? PMID- 11904943 TI - Why not infection control? PMID- 11904944 TI - Lessons from joy living with disability. PMID- 11904946 TI - Deciding to die. PMID- 11904945 TI - Seeking a compassionate death. PMID- 11904947 TI - Considering what is at stake. PMID- 11904948 TI - What the bible says about organ transplants. PMID- 11904949 TI - How small is too small? Issues surrounding the low-birth-weight infant. PMID- 11904950 TI - Kathy Beck. A ministry to moms in Ecuador. PMID- 11904951 TI - Stretching the boundaries. An ER dilemma. PMID- 11904952 TI - Caring without touching? PMID- 11904953 TI - Dissecting the dilemma. PMID- 11904954 TI - [Brief overview on the studies of monoclonal antibody]. AB - There have been many basic and clinical reports of monoclonal antibody therapies of cancer, but most of them were in the phase I or early phase II. However, from these experiences, monoclonal antibody therapies have changed considerably using chimera or humanized antibodies. Recent reports on monoclonal therapies indicated the clinical usefulness in cancer treatment, opening a new era of biological therapies. Since there are no similar issues to cover so widely monoclonal antibody, this special issue will be useful for understanding the progress of monoclonal therapies. PMID- 11904955 TI - [The expression technology of chimeric and humanized antibodies]. AB - Since the discovery of monoclonal antibody(MoAb) in 1975, MoAbs have held great promise for the treatment of human disease such as cancer, viral infection, and autoimmune disorders. The most important limitations in murine MoAb have been the immune response against murine immunoglobulins and insufficient activation of human effector function. The emergence of antibodies as an attractive therapy is the result of the evolution of MoAb technology over the past 25 years. These problems have been overcome using genetic engineering techniques to produce chimeric mouse-human, CDR-grafted, and fully human antibodies. New strategies to develop chimeric, humanized, and completely human antibodies are discussed in this review. PMID- 11904956 TI - [Exploring high affinity monoclonal antibodies for drug invention]. AB - Antibodies can have a medicinal role because of their high specificity in molecular recognition. Although monoclonal antibodies specific for an antigen can be obtained relatively easily, it is difficult to acquire ones with high affinity. Recently, antibody engineering using an in vitro evolutionary process has been put to this task. However, little progress seems to have been made. Our previous study revealed that antibodies have low or high evolvability. An antibody with a poor ability to evolve would also not attain high affinity in in vitro maturation. In this review, we provide information for the improved design of the antigen-combining site and that this will contribute to the engineering of antibodies using an in vitro evolutionary process. PMID- 11904957 TI - [Monoclonal antibody induces apoptosis against cancer cells]. AB - ErbB-2, a member of the epidermal growth factor(EGF) receptor tyrosine kinase family, is often overexpressed and/or amplified in breast, ovarian and gastric cancers, and other malignancies. ErbB-2 is a candidate as one of the best target molecules for cancer therapy. Many anti-ErbB-2 monoclonal antibodies(MoAbs) have been developed. An inhibitory humanized MoAb shows clinical responses in some breast cancer patients, both with MoAb alone and in combination with Cisplatinum or other anti-cancer drugs. A mouse-human chimeric anti-ErbB-2 MoAb CH401 was established and characterized in our laboratory. CH401 is able to kill cancer cells overexpressing ErbB-2 both in vitro and in vivo. The analysis of this tumor growth inhibition by CH401 made it clear that the cytotoxicity was induced by apoptosis. These results may suggest that CH401 has a therapeutic potential for ErbB-2 overexpressing cancers. This approach may be particularly valuable as a new type of cancer therapy. PMID- 11904958 TI - [Overview of recent monoclonal antibody therapy]. AB - It has been 27 years since Milstein and Kohler stunned the world of biology by announcing the discovery of monoclonal antibodies. Novel MoAb improved forms of chimeric and humanized antibodies with high antigen-binding affinities are currently being developed by both genetic and chemical methods. The rapid growth of molecular techniques will permit the development of novel antibodies, which should significantly improve conventional therapeutic strategies. This report overviews recent advances in the field of monoclonal antibodies and provides insight regarding the promises and limitations of this novel therapeutic approach. PMID- 11904959 TI - [Rationale for Herceptin in the clinical use]. AB - The human epidermal growth factor receptor-2(HER2) is overexpressed/amplified in a number of cancers. HER2 is implicated in disease initiation and progression and associated with poor prognosis. Trastuzumab(Herceptin) is a recombinant DNA derived humanized anti-HER2 monoclonal antibody that selectively binds with high affinity to extra-cellular domain. In vitro assay, trastuzumab has been shown to inhibit the proliferation of human tumor cells that overexpressed HER2 and to be a mediator of antibody-dependent cellular toxicity. Clinical trials have demonstrated that it is effective as both single and combination with chemotherapeutic agent in recurrent and metastatic breast cancer patients who's tumor tissue are overexpressed HER2. The incidence of severe adverse events were low but the occurrence of cardiac toxicity was unexpectedly high if trastuzumab was combined with anthracycline containing chemotherapy. The further studies are underway to assess the role of trastuzumab in combination chemotherapy, adjuvant chemotherapy and other type of tumors. PMID- 11904960 TI - [Rituximab, a chimeric mouse-human anti-CD20 monoclonal antibody]. AB - CD20 antigen is expressed on nearly all human B-cells and B-lymphoma cells. Rituximab is a chimeric anti-CD20 monoclonal antibody with mouse variable and human constant regions. The toxicities of rituximab are mainly infusion-related, non-hematological grade 1 or 2 episodes. Of the 11 eligible patients enrolled in the phase I study in Japan, 2 showed CR and 5 showed PR. 90 relapsed pts were enrolled in the subsequent phase II study and treated with rituximab at 375 mg/m2 x 4 weekly infusions. The overall response rates in relapsed indolent B-cell lymphoma and mantle cell lymphoma were 61%(37/61) and 46%(6/13), respectively. Rituximab is a novel, effective anti-lymphoma agent with acceptable toxicities. PMID- 11904961 TI - [Anti Shiga-like toxin II(SLT-II) humanized monoclonal antibody]. AB - Anti-Shiga-Like Toxin II(SLT-II) Humanized Monoclonal Antibody(TMA-15) was constructed from Mouse Monoclonal Antibody(MuVTm1.1) recognizing the same antigen using recombinant and CDR grafting technology. TMA-15 had almost the same affinity to SLT-II as MuVTm1.1 and showed the good protective activity of mice challenged either with SLT-II or with SLT-II secreting Shiga-like Toxin producing E. coli(STEC). TMA-15 showed no acute toxicity to monkeys and no cross-reactivity to human tissues in pre-clinical safety studies. From the preliminary results of Phase 1 clinical trial using healthy adult volunteers, doses up to planned maximum dose were well tolerated and TMA-15 showed long half life in blood almost comparable to gamma globulin preparations. Therefore, TMA-15 is expected to show clinical efficacy in coming clinical trial using pediatric STEC patients. PMID- 11904962 TI - [Infliximab(chimeric monoclonal antibody to tumor necrosis factor alpha)]. AB - In rheumatoid arthritis and inflammatory bowel disease, such as Crohn's disease, certain immunological abnormalities are considered as its cause, but the fundamental mechanism remains unclear. However, recent researches revealed that inflammatory cytokines, such as TNF-alpha, IL-1 and IL-6, are greatly involved in these immunological abnormalities. This led to the development of anti-cytokine therapy using monoclonal antibodies to these cytokines. Among them, Infliximab, a chimeric monoclonal antibodies to TNF-alpha, is most clinically applied and marked therapeutic effect is seen in various diseases. PMID- 11904963 TI - [Humanized respiratory syncytial virus monoclonal antibody]. AB - Respiratory syncytial virus remains a significant cause of severe lower respiratory tract disease in children. The risk of serious RSV illness is highest among children with prematurity, chronic lung disease and congenital heart disease. No effective vaccine and anti-viral agents have been obtained even now. Therefore, conservative therapy including respiratory aid has been a principal therapy for serious RSV disease. Recently, monthly intramuscular administration of humanized anti-RSV monoclonal antibody(palivizumab) was introduced in clinical fields in USA and Europe. Palivizumab prophylaxis has appeared to be safe and effective for prevention of serious RSV illness in premature children and those with bronchopulmonary dysplasia. PMID- 11904965 TI - [Monoclonal immunotherapy with human monoclonal antibody(CLN-IgG) in glioma patients]. AB - It is well recognized that malignant gliomas escape an immune response by hiding behind the blood-brain barrier and by producing proteins that suppress systemic immunity. However, if gliomas can be made to be more immunogenic or if a tumor vaccine can be produced, then access to all tumor cells including those that infiltrate into the brain can be achieved through the patient's immune response. Several strategies have been investigated for immunotherapy. Laboratory studies and animal models have shown that these immune cells will attack the tumor cell, reduce the size of implanted tumors, and that the immune memory is sufficient to suppress tumor growth when the animal is rechallenges with a tumor implant. Since the development of hybridoma technology, monoclonal antibodies against human cancer cells have been produced and antigens have been identified. Hagiwara reported the production of a human monoclonal antibody, CLN-IgG, made by fusing UC 729-6, human lymphoblastoid B-cell line, with lymphocytes obtained from a patient with the cervical carcinoma. It has been reported that CLN-IgG recognized the antigen expressed in various histological types of human cancers including malignant gliomas. The effect of human monoclonal antibody(CLN-IgG) on malignant brain tumors was evaluated in patients with malignant glioma. Early phase II study was concluded that this specific immunotherapy with CLN-IgG is safe and effective therapy in patients with malignant glioma. We treated 10 cases of malignant gliomas with CLN-IgG. All patients had received radiotherapy and chemotherapy before this immunotherapy using the human monoclonal antibody. The human monoclonal antibody(CLN-IgG) was administered intravenously once or twice/week during 24 weeks. Six cases of glioblastoma, 1 medulloblastoma and 3 cases of potine glioma histologically unverified, were treated. Five cases of 6 glioblastomas died 4 to 12 months after this treatment, 3 cases of pontine glioma showed good responses, 2 cases showed marked decrease of tumor size and 1 case showed no regrowth of tumor on MRI imaging. For the above reasons, Human monoclonal antibody(CLN-IgG) might be useful as an immunotherapy of malignant gliomas. PMID- 11904964 TI - [New therapy with monoclonal antibodies--abciximab, chimeric anti-GP IIb/IIIa antibody]. AB - Platelet aggregation is now believed to be mediated by fibrinogen binding to activated GP IIb/IIIa. Antigen binding site of mouse anti-human GP IIb/IIIa monoclonal antibody 7E3, which can inhibit fibrinogen binding to activated human platelet GP IIb/IIIa, was combined with constant region of human IgG to reduce immunogenicity. Monovalent Fab of this mouse-human chimeric antibody named abciximab effectively reduced the onset of cardiovascular accidents when administered in combination with heparin and aspirin in patients undergoing coronary interventions. Although several anti-GP IIb/IIIa other than monoclonal antibodies were developed, clinical efficiency of abciximab is still superior to other agents while its effects on preventing myocardial infarction in unstable angina patients are uncertain. PMID- 11904966 TI - [Development of monoclonal antibody therapy for malignant lymphoma]. AB - Lymphomas are among the tumors most responsive to chemotherapeutic agents and radiation therapy. Despite chemotherapeutic and radiological advances, tumor cell resistance still remains a problem, and the toxicity of chemotherapy and radiation therapy limits their potential. Less toxic therapies for lymphoma have been continued to search for effectiveness. Since the discovery of the hybridoma technology by Kohler and Milstein in 1975, utilizing antibodies as targeted therapy for lymphoma has been investigated for many years. After 20 years of clinical trials, monoclonal antibody therapy of lymphoma enters the new millennium ready for 'prime time'. Investigators' early enthusiasm was dampened by problems with tumor targeting, HAMA, and allergic reactions, but important advances in molecular biology and chelation chemistry have led to new and improved reagents. Rituximab(IDEC-C2B8) has already been approved by the FDA in USA and the Ministry of Welfare and Labour in Japan for relapsed CD20-positive lymphomas and indolent B-cell lymphoma including mantle cell lymphoma, respectively. Ibritumomab tiuxetan and iodine-131 anti-B1 antibody have an excellent anti-lymphoma profile, and both appear to have higher response rates than rituximab. Results from the rituximab vs. ibritumomab tiuxetan phase III trial clearly favor the latter especially in %CR. Radiolabeled Lym-1, T101, LL2, and anti-Tac data will be forthcoming. Continued refinements of immunotoxins will establish their possible therapeutic role, and a variety of antibody conjugates including drugs, prodrugs, nonprotein toxins, and other agents, will continue to be studied in the clinic. Bispecific antibodies for lymphoma are also in early clinical testing. Over the next 10 years, many of the major advances in lymphoma therapy will be antibody-based. PMID- 11904967 TI - [Antibody directed therapy for leukemia]. AB - Recently, monoclonal antibody(MoAb) therapies which direct at antigens such as CD33, CD45 and GM-CSF receptors on myeloid leukemia cells have been in progress. There are three major MoAb therapies against acute myeloid leukemia(AML), which include unconjugated MoAb, MoAb conjugated with chemotherapy or toxins, and MoAb conjugated with radioisotopes. Gemtuzumab ozogamicin is consisting of an engineered human anti-CD33 antibody linked with the potent anti-tumor antibiotic calicheamicin, which is the most effective for AML. However, recent studies suggested that calicheamicin was also pumped out by multi-drug resistance(MDR) related P-glycoprotein. The combination therapy with MDR modifiers may improve the effect of gemtuzumab ozogamicin. PMID- 11904968 TI - [Monoclonal antibody therapy for disorders of hemostasis and coagulation]. AB - Monoclonal antibody therapies have conducted to not only hematologic malignancies but also disorders of hemostasis and coagulation. This article describes the recent advances of monoclonal antibody therapy for bleeding disorders such as idiopathic thrombocytopenic purpura(ITP), hemophilia A, disseminated intravascular coagulation(DIC), and thrombosis. Rituximab, chimeric anti-CD20 monoclonal antibody treatment has a valuable effect in the patients with ITP, and clinical trials using anti-CD40 ligand monoclonal antibody for ITP are underway. Anti-CD40 ligand monoclonal antibody can be an alternative therapy for hemophilia A patients with inhibitors to factor VIII. In thrombosis, anti-tissue factor monoclonal antibody and anti-factor IX(a) monoclonal antibody were established as novel anticoagulant regents. Plasminogen activator inhibitor-1(PAI-1) increases in endotoxin-induced DIC and many thrombotic diseases such as myocardial infarction, type 2 diabetes mellitus, and hyperlipidemia. Anti-PAI-1 monoclonal antibody reduced fibrin deposition in DIC mouse model. Treatment of these monoclonal antibodies for the molecules regulating coagulation-fibrinolysis system may be utilized for acute coronary syndrome and venous thrombosis. PMID- 11904969 TI - [Treatment of ulcerative colitis and Crohn's disease with monoclonal antibody]. AB - Ulcerative colitis and Crohn's disease are nonspecific inflammatory diseases of unknown etiology. Recent immunological studies have shown that proinflammatory cytokines and adhesion molecules play an important role in the pathogenesis of ulcerative colitis and Crohn's disease. Therefore, monoclonal antibodies to proinflammatory cytokines and adhesion molecules are used to suppress the mucosal inflammatory response in experimental colitis and ulcerative colitis and Crohn's disease. Anti-TNF alpha antibody and anti-alpha 4 beta 7 integrin antibody are well-tolerated and effective for treatment of patients with Crohn's disease. This review described clinical features and immunopathophysiology of ulcerative colitis and Crohn's disease, proinflammatory cytokines and immunosuppressive cytokines and adhesion molecules involved in the pathogenesis of both disease, and treatment of both diseases with monoclonal antibodies. PMID- 11904970 TI - [Clinical assessment of monoclonal antibodies for the treatment and diagnosis of colorectal cancers]. AB - The clinically useful monoclonal antibodies(Mabs) for colorectal cancers were reviewed. Since 1980's, immunoscintigraphy has been performed for the detection of occult colorectal cancers. However, it may be substituted with the development of positron emission tomography. As for the treatment, some Mabs have been shown to be effective for the adjuvant therapy of postoperative colorectal cancers. Some Mabs to epidermal growth factor receptors(EGFr) are quite promising since they block the functions necessary for the tumor growth and enhance the cytotoxicity of chemotherapy. Recent advances for the development of humanized Mabs will improve the chance of Mabs to be used as an effective adjuvant. PMID- 11904971 TI - [Shiga toxin producing Escherichia coli infection]. AB - Shiga toxin producing Escherichia coli(STEC) has been recognized as an emerging food-borne pathogen that causes bloody diarrhea, hemorrhagic colitis and hemolytic uremic syndrome(HUS), especially in developed countries. As the specific therapy for STEC infection has not been developed, currently available medical therapy is inadequate to prevent life-threatening complications. Here are described the possibilities and problems of using and developing therapies such as antibiotics, Synsorb-Pk and humanized anti-Stx monoclonal antibody therapy. In conclusion, the prevention of primary infection is thought to be the best way to prevent the life-threatening complications caused by STEC, and second way is identification as early as possible. PMID- 11904973 TI - [Monoclonal antibody therapy for uveitis]. AB - Uveitis, or intraocular inflammation, is caused by various reasons. Since many of the cases are of unknown origin, non-specific anti-inflammatory therapy is often the only choice for the treatment of uveitis. Furthermore, some of the patients are indeed refractory to every available modality and are suffering from severe visual disturbances. Stronger medicine is needed. Monoclonal antibodies against tumor necrosis factor-alpha, TNF-alpha, are one of the candidates in giving us more opportunities to treat such patients more effectively. We are especially hopeful of its application to refractory Behcet's disease. Its unique adverse side effects, though, should be carefully monitored. On balance, this monoclonal antibody therapy may become the last anchor for treating sight-threatening uveitis in the near future. PMID- 11904974 TI - [Breast cancer]. AB - The HER2/neu protein is thought to be a unique and useful target for antibody therapy of cancers overexpressing the HER2/neu gene. The recombinant humanized anti-HER2 monoclonal antibody, trastuzumab(Herceptin) has recently been available for clinical use in Japan. In this paper, the details of this novel biologic agent are reviewed in conjunction with the results of the clinical trials for breast cancer. PMID- 11904972 TI - [Monoclonal antibody therapy for rheumatoid arthritis]. AB - Rheumatoid arthritis(RA) is a systemic chronic inflammatory disease characterized by destructive polyarthritis. Although its etiology and pathogenesis are still to be elucidated, recent cumulative evidence has suggested both T cells and proinflammatory cytokines (such as tumor necrosis factor(TNF)-alpha, interleukin(IL)-1, IL-6) play pivotal roles in the pathogenesis. Therefore, they are thought to be optimal therapeutic targets. In fact, new therapies targeted to T cells and proinflammatory cytokines using biological agents including monoclonal antibodies have been developed extensively. Among them, anti proinflammatory cytokine therapies, such as anti-TNF-alpha therapy, have shown an excellent efficacy and some are currently accepted as a new promising therapy for RA. It is expected that further investigation will provide additional insights into cure of RA. PMID- 11904975 TI - [Human monoclonal antibodies to HIV-1 infection]. AB - HIV-1, which causes AIDS, has infected over 50 million and killed over 20 million individuals world wide since the beginning of the epidemic two decades ago. The introduction of highly active anti-retroviral therapy(HAART) has resulted in the control of the disease progression. Furthermore, supervised treatment interruption(STI) of HAART might be expected significantly to boost immune function and slow the progression of AIDS. On the other hand, several combinations of human monoclonal antibodies(hu-mo-Ab) against HIV-1 has been identified, and demonstrates the efficacy of the triple combination of hu-mo-Ab with neutralizing activity in macaque model. Therefore, a combination of chemotherapy and immunotherapy, such as hu-mo-Ab will be prove to be the most effective route for HIV-1 therapy. PMID- 11904976 TI - [Monoclonal antibodies against inflammatory mediators for the treatment of patients with sepsis]. AB - Sepsis is a common cause of morbidity and mortality, particularly in immunocompromised and critically ill patients. Recently, a new designation, systemic inflammatory response syndrome(SIRS), has been studied. When an abnormal generalized inflammatory reaction is due to an infection, the terms sepsis and SIRS are synonymous. The systemic response to infection is mediated via the macrophage-derived cytokines that target end organ receptors in response to injury or infection. One strategy used to perturb the septic cascade is to block a particular inflammatory molecule. Results have been published on clinical trials in sepsis patients treated with several monoclonal antibodies, such as antiendotoxin antibodies, anti-tumor necrosis factor antibodies, and anti CD14 antibodies. In this chapter, the results of clinical trials in patients and in vivo data from animal models of sepsis are summarized. PMID- 11904977 TI - [Viral infection]. AB - Viral infections, particularly in the fetus or newborn and in immunocompromised individuals, can produce severe, even fatal disease. Indeed Herpesviridae infections are the major cause of morbidity and mortality in them. Antiviral drugs have been used for prophylaxis and treatment for some viral infections. An alternative approach is to boost the host immune defences. Administration of pooled human sera have been used for prophylaxis or alleviation of clinical severity of ongoing infections with measles, CMV, VZV and hepatitis A and B and so on. Human monoclonal antibody preparations, which have now being developed, may overcome the problems inherent in the pooled sera. These include high volume load, difficulty of quality control and adventitious transmission of unknown pathogenic agents. PMID- 11904978 TI - [Monoclonal antibody therapy for allergic asthma]. AB - Allergic responses at the level of the respiratory system are mostly mediated by IgE-dependent mechanisms. The first selective anti-IgE therapy, a recombinant humanized monoclonal anti-IgE antibody(rhuMAb-E25), binds with high affinity to the Fc epsilon RI receptor binding site on IgE, thereby reducing the amount of free IgE available to bind to Fc epsilon RI receptors on mast cells and basophils. In addition, administration of rhuMAb-E25 indirectly reduces Fc epsilon RI receptor density on cells involved in allergic responses. rhuMAb-E25 has been shown to reduce allergic responses in atopic individuals and to improve symptoms and reduce rescue medication and corticosteroid use in patient with allergic asthma. The clinical effectiveness of rhuMAb-E25 supports the central role of IgE in allergic reaction and the viability of anti-IgE therapy as an effective immunological intervention for allergic asthma. PMID- 11904979 TI - [Therapeutic strategy for acute rejection in organ transplantation--optimal use of monoclonal antibody]. AB - Transplantation has now become the treatment of choice for end-stage organ failure. However, acute rejection still occurs in approximately 20-50% of cadaver or living donor kidney recipients during the first year post-transplantation and lead to permanent damage of graft function or loss. Recently chimeric(basiliximab) and the humanized(daclizumab) monoclonal antibodies against the alpha chain(CD25) of the interleukin 2 receptor(IL-2R) introduced in the late 1990s. Induction therapy with these antibodies results in a 15-20% reduction in acute rejection episodes after renal transplantation. The availability of anti-IL 2R monoclonal antibodies offers a further opportunity for graft-specific induction therapy. PMID- 11904980 TI - [Genetic testing for effective Herceptin therapy]. AB - Herceptin has provided the first proof that tyrosine kinase modulation, through monoclonal antibodies can translate into improved clinical outcomes in cancer therapy. The development of Herceptin was encouraged by the biologic significance of HER2 overexpression. Although the number of patients affected by the targeted molecular abnormality(30% of breast cancer patients) is small and the response rate observed in patients after treated with single agent Herceptin is rather low, the ability to document overexpression of the target in breast biopsies, and a growing interest in biologic therapy facilitated the rapid accrual of patients to clinical trials. The challenges of applying research techniques of molecular biology to routine clinical testing have been demonstrated by the experiences with the HER2/neu oncogene. Immunohistochemistry and fluorescence in situ hybridization yielded discrepant results regarding the frequency and degree of HER2 alterations even within the same sample. In order to make the complexity of interpretation of the discrepancy simple, it is wise to build an algorithm to help clinicians follow the ideal sequence of laboratory testings. The experience gained in the testing of Herceptin has provided important lessons for the future testing of molecularly targeted compounds. PMID- 11904981 TI - [Heterogeneity of fibrosis in the lungs]. AB - Fibrosis in the lungs has been conceptually summarized into 5 types by Spain D in 1950 when pathologists examined them using autopsy specimens. However, interstitial, parenchymal, bronchiolar, vascular and pleural fibrosis were presented differently. Since then, several critical issues can be presented in order to elucidate more effective therapeutic strategies in the areas of lung fibrosis. One is the spectrum of interstitial pneumonias, and other is a peribronchiolar pulmonary fibrosis with distortion of bronchos which is different from the type of interstitial pneumonia. Usual interstitial pneumonia(UIP) and fibrosis of sarcoidosis are two typical representative which are warranted for a development of effective therapy. Based on these recognition, a significance of pulmonary fibrosis intensively examined using animal model and several clinical research are reviewed. PMID- 11904982 TI - [Non surgical new treatment modalities for lung cancer]. PMID- 11904983 TI - [Developed new agents for lung cancer]. AB - Platinum-based chemotherapy is considered to be the standard chemotherapy of non small cell lung cancer (NSCLC) at present. New agents such as irinotecan, paclitaxel, docetaxel, vinorelbine, gemcitabine, topotecan, and amurubicin were developed in the 1990s. Combination chemotherapy using new agents improved survival rates compared with the classic regimen. Irinotecan, paclitaxel, docetaxel, vinorelbine, and gemcitabine have been confirmed to be effective against NSCLC. However, chemotherapy for NSCLC is controversial because the differences in the efficacies of combination chemotherapies including new agents have not been recognized in randomized controlled trials. The Four-Arms Cooperative Study is an ongoing postmarketing clinical trial in Japan. The a ntitumor agents irinotecan, topotecan, paclitaxel, and amurubicin have been confirmed to be effective in small cell lung cancer (SCLC). The combination of etoposide and cisplatin (PE) is the standard regimen for SCLC in Western countries. However, the combination of irinotecan and cisplatin (CP) resulted in higher response rates and better median survival times than PE for extensive disease SCLC in a JCOG trial. At present, CP is considered to be the standard chemotherapy regimen to treat SCLC in Japan. The development of new agents, particularly molecular target-based drugs and multimodality treatment, is necessary to improve the therapeutic results in lung cancer further. PMID- 11904984 TI - [Drug delivery systems for cancer chemotherapy]. AB - Various types of drug delivery system (DDS) for antitumor drugs have been developed to reduce severe systemic toxicities and to enhance antitumor effects by improving their pharmacokinetics. In this paper, outlines are first given of the concept and design of DDS, focusing on polymeric antitumor drugs (macromolecular prodrugs) and then describing the superior DDS effects shown in preclinical studies of DE-310, a DDS optimized for the camptothecin analogue DX 8951. Clinical trials of DE-310 are now ongoing in the USA and Europe. PMID- 11904985 TI - [Chemosensitivity test for unresectable non-small cell lung cancer]. AB - To choose the optimal chemotherapy regimens, we have employed a new chemosensitivity testing method, the collagen gel droplet embedded culture drug sensitivity test (CDDST) for patients with non-small cell lung cancer (NSCLC). This method requires fewer cancer cells(1 x 10(5) cells in 2 specimens biopsied by bronchoscopy) than conventional chemosensitivity tests and can be also used to assess cases of malignant effusion. Correlations between the in vitro and in vivo responses were: true positive ratio, 75.0% (21/28 patients); true negative ratio 85.0% (17/20 patients); and accuracy 79.2%. The median survival time (MST) of patients (n = 11) with unresectable NSCLC who were given optimal chemotherapy based on the results of the CDDST was 15.8 months and the MST of those (n = 16) who did not receive a sensitive agent was 5.6 months. There was a significant difference between these two groups (p = 0.0048, log-rank test). These results suggest that the CDDST is an effective method for chemosensitivity testing in unresectable NSCLC. PMID- 11904986 TI - [Molecular target-based cancer therapy: epidermal growth factor receptor inhibitors]. AB - Based on recent progress in cancer biology, numerous molecules that contribute to proliferation, invasion, and metastasis of cancer cells have been identified. The epidermal growth factor receptor (EGFR), a member of cell membrane receptors, is overexpressed by many tumors, and EGFR overexpression correlates with poor prognosis and disease progression. The EGFR is an attractive target for novel anticancer therapy. ZD1839 and OSI-774, highly specific EGFR tyrosine kinase inhibitors, have shown promising antitumor activity against cisplatin-resistant non-small cell lung cancer in phase I and phase II trials. IMC-C225, a monoclonal antibody against EGFR, has achieved significant disease control in head and neck cancer and colorectal cancer in combination with anticancer agents. These agents are under evaluation in phase III trials. In conclusion, it is expected that EGFR directed therapies will soon be established as an effective novel treatment for many cancer patients. PMID- 11904987 TI - [Development of antiangiogenic chemotherapy regimens]. AB - Research in tumor angiogenesis is making rapid progress. It was recently confirmed that tumors utilize both vasculogenesis and existing blood vessels for angiogenesis, and angiogenesis has become a molecular target of new cancer therapies. Numerous inhibitors of angiogenesis have been developed and clinical trials conducted. However, problems such as side effects and the appropriate design of clinical trials have been noted, and the development of some inhibitors has stalled. Therefore more time and study will be required before clinical application of angiogenesis inhibitors is feasible. Trials combining existing chemotherapeutic regimens with angiogenesis inhibitors is feasible. Trials combining existing chemotherapeutic regimens with angiogenesis inhibitors, i.e., antiangiogenic chemotherapy, are underway in which low-dose anticancer and antiangiogenic agents are administered daily over the long term, and the results are awaited. PMID- 11904988 TI - [Gene therapy for lung cancer]. AB - Surgical resection is so far the most reliable therapy for localized non-small cell lung cancer (NSCLC). Although the combination of several modalities such as chemotherapy or radiotherapy is used to treat advanced or recurrent disease, the results have not been satisfactory. A new therapeutic strategy is thus required to improve lung cancer treatment. Gene therapy is one such new therapeutic strategy. Several clinical trials of gene therapy protocols have been conducted and patient eligibility and being efficacy are evaluated. Representative studies of gene therapy for lung cancer and an interim report on a clinical trial using adenovirus vector expressing wild-type p53 in non-small cell lung cancer in Japan are introduced in this paper. Problems encountered and future regimens are also discussed. PMID- 11904990 TI - [Stereotactic radiation therapy for non small cell lung cancer]. AB - Stereotactic radiation therapy (SRT) is a highly effective treatment for brain metastases from non-small cell lung cancer (NSCLC). Primary lesions of NSCLC may also be controlled by SRT. Between 1994 and 1999, 50 patients with pathologically confirmed T1-2N0 M0 NSCLC were treated with SRT. With a median follow-up period of 36 months, 3-year cause-specific survival was 88%. SRT is a very safe and effective treatment for early stage NSCLC. PMID- 11904989 TI - [Heavy-ion therapy for non-small cell lung cancer]. AB - Since carbon beam therapy for non-small cell lung cancer (NSCLC) was initiated in October 1996, seven trials have been conducted; three have already closed and the remaining four are ongoing. The local control rate, cause-specific survival rate, and overall survival rate of 141 patients with clinical stage I NSCLC were 82%, 58%, and 42%, respectively. Radiation pneumonia was rare (2.1%) and not serious. In the phase II clinical study, the local control rate achieved in 50 patients was 100%, with no radiation pneumonia, resulting in a 60% overall survival rate. Carbon beam therapy could be an alternative to surgery, especially for lung cancer patients of advanced age and/or with complications. For locally advanced lung cancer treated with carbon beam therapy, excellent local control comparable to that in stage INSCLC has been demonstrated and offers hopeful prospects for the treatment of lung cancer. PMID- 11904991 TI - [Photodynamic therapy for lung cancer: state of the art and expanded indications]. AB - Photodynamic therapy (PDT) has now achieved the status of a standard treatment modality for centrally located, early-stage lung cancer and is introduced on the home page of the US National Cancer Institute. Over the past decade, 145 patients (191 lesions) with central-type early-stage lung cancers have been treated with PDT in our hospital. Overall complete remission was obtained in 86.4% of the total number of lesions. In patients with advanced stage III disease, however, bronchi opening was successful in 61 out of 81 lesions (75%) for the PDT group compared with 143 of 177 (81%) in the Nd-YAG laser therapy group. Although quite recent, treatment using PDT has been introduced for the first time in patients with peripheral lung cancer, who did not previously meet the criteria for surgery. Eight patients underwent this trial, of which 3 achieved partial remission. No serious complications except for one case of pneumothorax, were noted. As increasing number of patients consider quality of life after therapy, the indications for PDT are expected to expand. The success in clinical trials of PDT for cancer treatment offers encouragement for its future use. PMID- 11904992 TI - [My diary in Dandeldhura Hospital]. PMID- 11904993 TI - [On the origin of naming "pancreas"]. PMID- 11904994 TI - [An estimation of imported infections concerning 2002 FIFA world cup Korea/Japan]. AB - The FIFA World Cup 2002 Korea/Japan, which will be held during May 31 through June 30, 2002, is a mass-gathering and high-profile event. The Ministry of Transport announced that approximately 430,000 people visit Japan for the event. We estimated the incidence of major imported infections using data from the national epidemiological surveillance of infectious diseases and the statistics of immigration. Estimated incidences are 5.88 (shigellosis), 3.41 (malaria), 1.40 (typhoid/paratyphoid fever), 0.42 (cholera), and 0.0032 (meningococcal meningitis). The incidence for viral hemorrhagic fever was estimated 0.0018 under assumption that "it correlates with the malaria incidence from Africa" and that "the incidence occurs every 15 years". These results indicate little possibility of remarkable increase of exotic infections during the event. These incidences, however, may occur in the rural prefectures where few cases are reported. It is highly needed to strengthen surveillance and educate physicians and public health experts especially for malaria cases. PMID- 11904995 TI - [Community acquired sepsis by Serratia rubidaea]. AB - A 48-year-old male who had a past history of alcoholic pancreatitis and diabetes mellitus was admitted to our hospital due to chills and vomiting, on August 13, 1998. His body temperature was 38.0 degrees C, and he had the disturbance of consciousness, tachypnea, tachycardia and hepatomegaly with tenderness. Laboratory findings showed highly inflammatory reactions, DIC and hepatorenal dysfunction. Abdominal CT and US revealed multiple liver abscess with portal vein thrombus. Serratia rubidaea was detected in the blood culture. SBT/CPZ and TOB were administered and he recovered. This is a rare case of Serratia rubidaea sepsis. It is also necessary to pay attention to Serratia infections as well as S. marcescens. PMID- 11904996 TI - [A case of cat scratch disease with encephalopathy]. AB - We report an atypical case of cat scratch disease (CSD), accompanied with encephalopathy that is a rare complication of CSD. A 17-year old man consulted a doctor for his right axillary lymphadenopathy. The history of his contact with cats and the sign of lymphnode swelling and fever suggested a suspect of cat scratch disease. Administration of ampicillin improved his clinical symptoms, but a few days later he suddenly fell into coma after an episode of convulsion. The CT scan of the brain and laboratory tests showed no significant findings except the slightly elevated cell counts and concentration of protein in his cerebrospinal fluid. He was referred to our hospital on the next day for further examinations and treatments for his coma of unknown cause. The physical examination on admission revealed slight neck stiffening and hypertonicity of his right lower limb, but radiological and laboratory tests showed no significant findings. He gradually recovered from his coma without apparent sequelae in three weeks. Indirect fluorescence antibody titers for CSD in his serum showed a significant elevation to 1:160 of IgM and 1:512 of IgG, and his clinical features were compatible to these of CSD with complications of the central nervous system. PMID- 11904997 TI - An imported case of cyclosporiasis in Japan. PMID- 11904998 TI - [Epidemiological interference between influenza-like illness and respiratory syncytial virus infection in children]. PMID- 11904999 TI - [Seroepidemiology of Coxiella burnetii in Okayama Prefecture, Japan]. PMID- 11905000 TI - [Protective effect of azithromycin against verotoxin]. PMID- 11905001 TI - [Current topics on classification and nomenclature of bacteria 10. Nomenclature and its social influence]. PMID- 11905002 TI - [Clinical and bacteriological features of six cases with intracranial abscess in childhood]. AB - From October 1988 to March 2001, 5 patients with 6 episodes of intracranial abscesses were admitted to Chiba-Children's Hospital. Average age when they were admitted was 10 years and 1 month. Initial clinical symptoms were fever in 5 cases, and headache in 1 case. It took 21 days from the appearance of the initial symptoms to diagnose the intracranial abscess. Four out of 5 patients had underlying diseases that were prone to cause intracranial abscess. Two patients of these were cyanotic congenital heart diseases (tetralogy of Fallot and asplenic heart), and the other 2 were sinusitis. Computed tomography revealed that brain abscess was found in 5 cases, and subdural empyema in 1 case. There were 3 single and multiple abscesses each. The most common lesion was the temporal lobe. Eight bacterial strains were isolated from 5 cases. Five were streptococci (3 were Streptococcus milleri group, other 2 were Streptococcus oralis and microaerophilic Streptococcus) and 3 were anaerobes (Prevotella loescheii, Prevotella bivia and Fusobacterium nucleatum). Antimicrobial therapy was started with panipenem-betamiprone in 3 cases, imipenem-cilastatin, ceftriaxone, and ampicillin in the other cases resre ctinely. Duration of therapy ranged from 28 to 67 days (45 days, average). In 5 cases, drainage with craniotomy was performed in addition to antimicrobial therapy. One case was treated medically alone, but this was the only case with recurrence after 1 year 2 months. There were no serious complications such as intraventricular rupture of abscess. All patients had good outcomes, but mild neurological sequela was found in 1 case. PMID- 11905003 TI - [Investigate of nasopharyngeal flora in highly aged patients]. AB - To clarify the bacteriological interpretation of flora in the nasopharynx of highly aged patients (n = 107), healthy nasopharyngeal swabs were obtained from subjects of advanced age. Chief pathogenic bacteria isolated from highly aged persons were coagulase negative Staphylococcus (43 strains), Corynebacterium spp. (14 strains), methicillin-susceptible Staphylococcus aureus (beta-lactamase production and non-production) (16 strains), methicillin-resistant Staphylococcus aureus (beta-lactamase production) (6 strains). Chief nonpathogenic bacteria isolated from highly aged persons were alpha-streptococcus (14 strains), Neisseria sp (3 strains). Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis which are the chief bacteria isolated from children, demonstrated only 3 strains for each bacteria. In cases showing detection of multiple detected bacteria, common combinations were non-pathogenic bacteria and weakly pathogenic bacteria and enteric bacteria and pathogenic bacteria. The differences between nasopharyngeal flora of children and highly aged persons are suspected to be due to the differences in immunological and anatomical factors. We should actively examine these factors in highly aged subjects. PMID- 11905004 TI - [Molecular epidemiological investigation of vero toxin-producing Escherichia coli (VTEC) isolated from diarrhea patients and dairy cattle]. AB - To elucidate the source and route of VTEC infection, we performed pulse field gel electrophoresis (PFGE) an 50 isolates from human diarrhea typed as serotypes O157, O111, and O26, which were very frequently isolated from patients with VTEC infection between 1986 and 1997, and 32 isolates from dairy cattle, a total of 82 isolates. The isolates were genetically analyzed based on the electrophoresis patterns of DNA, and a phylogenetic tree was prepared. The isolates were classified based on similarity > or = 89. The following results of the molecular epidemiological investigation were obtained. 1) Based on the electrophoresis patterns of DNA obtained by PFGE, 34 of the 49 O157 isolates (69.4%) were divided into groups 1-9, 15 of the 18 O111 isolates (83.3%) were divided into groups 1-3, and 12 of the 15 O26 isolates (80%) were divided into groups 1-3. Of the grouped isolates, group 8 of O157, groups 2 and 3 of O111, and group 3 of O26 included isolates from human diarrhea and dairy cattle, but the other groups included isolates from only one of the two sources. 2) With regard to regional investigation, groups 6 and 9 of O157 included human diarrhea-derived isolates from Yokohama and Ehime, and group 8 included a human diarrhea-derived isolate from Yokohama and a dairy cattle-derived isolate from Tokushima. Group 3 of O111 included a human diarrhea-derived isolate from Ehime and a dairy cattle-derived isolate from Hokkaido. Group 3 of O26 included human diarrhea-derived isolate from Ehime and dairy cattle-derived isolate from Sagamihara and Hokkaido. Since the above findings showed that although the frequency was low, isolates from human diarrhea and dairy cattle were included in the same groups, it was demonstrated that dairy cattle are closely related to the human infectious disease of the intestinal tract as a source of infection. However, classification using the PFGE method is difficult due to diversity of the electrophoresis pattern of DNA. It is necessary to investigate the classification by a combination of the PFGE method with phage typing, ribotyping, and RAPD-PCR, and to investigate more numbers of patient-derived and animal-derived isolates. PMID- 11905005 TI - [Social welfare for children with developmental disabilities in the 21st century]. AB - For children with developmental disabilities in Japan, various social policies including education, medical care, pension and welfare have rapidly developed and prevailed during the last 50 years. The concept of living together with the disabled has been widely accepted. A high standard of social welfare has been established by virtue of high respect for human rights, progress in medicine and economic growth. Historically, the International Year for the Disabled played an important role in meeting the expectation of the disabled and their families. Amendment of the legal system has realized normalization and social independence. Based on this historical background, there are in the 21st century new trends of development in the education, welfare and social life of the disabled, aiming at a better society for all the people. PMID- 11905006 TI - [Clinical application of functional magnetic resonance imaging]. AB - Three types of researches have been carried out on brain-mind relationships: 1. researches on anatomical correlates of special talents (for example, perfect pitch) or deficits (for example, dyslexia), 2. researches to examine the relationship between a given cognitive syndrome and the site of brain damage, 3. researches to localize human cognitive function in the brain in vivo using functional magnetic resonance imaging (fMRI) and positron emission tomography (PET). fMRI is a particularly important because it is noninvasive. A tutorial covering basic aspects of this methodology is presented, along with a survey of recent fMRI data related to clinical application. Future investigations of the three types enumerated above are expected to further clarify brain-mind relationships. PMID- 11905007 TI - [Neonatal neurology in extremely and very low birth weight infants: introductory remarks]. AB - Survival rates of extremely and very low birth weight infants have been improved dramatically. However, some of the survivors suffer from neurological sequelae. A recent report showed that about 25% of the children with birth weight of less than 1,000 g had some handicaps, including cerebral palsy, mental retardation, and visual disturbance. Therefore, it is very important for pediatric neurologists to assess the neurological symptoms properly in the neonatal period. This symposium was organized to present recent progress in the methods for neurological assessment and to discuss how we should support the development of high risk infants. PMID- 11905008 TI - [Clinical significance of general movements]. AB - Human fetuses and young infants have a repertoire of distinct patterns of spontaneous movements. A set of these movement patterns are known as general movements (GMs), which were defined by Prechtl as gross movements involving the whole body, and lasting from a few seconds to several minutes, or longer. GMs are characterized by the variable sequence of arm, leg, neck and trunk movements which begin gradually, wax and wane in intensity, force and speed, and end gradually. Extension and flexion movements of the arms and legs are mostly complex and variable because of superimposed rotations and frequent, slight changes in direction, which make the movements fluent and elegant. Initially GMs are complex, and then differentiate into single movements. GMs show no change during the fetal period, but change in early infancy into writhing, fidgety, oscillating, saccadic and swipes & swaps. Hadders-Algra indicated that on EMG, burst duration of phasic activity shortens, burst amplitude attenuates and tonic background decreases with development. In contrast to normal GMs that are smaller and monotonous in trajectory, abnormal GMs include poor repertoire of GMs, cramped-synchronized GMs, chaotic GMs, absent fidgety, and abnormal fidgety, that are periodic and monotonous, showing no developmental change. For example, a normal twin showed elliptic or round trajectory of GMs that became smaller with age, whereas the other with periventricular leukomalacia and cerebral palsy had periodic and rectilinear trajectory showing no developmental change. GMs, especially fidgety, correlate with neurological prognosis of infants. Einspieler reported that GMs' sensitivity and specificity for neurological prognosis of high risk infants are 96% and 95%, respectively. Clinical significance of GMs 1. includes non-invasive, secure and easy observation, 2. high coincidence between trained observers, 3. high reliability 78-98%, mean 90% and, 4. correlation of abnormal GMs with the presence and degree of brain damage. PMID- 11905009 TI - [Neurological signs and assessment of extremely and very low birth weight infants]. AB - The infants' brain during the prenatal, perinatal and neonatal periods is susceptible to injury. Many problems in the perinatal period often result in bleeding, ischemia and other pathological changes in the infant brain. Which can subsequently cause cerebral palsy or developmental disorders. Unless they are discovered early and measures are taken, permanent brain damage may remain. Although neurological examinations at this stage is very difficult, it is very important to be familiar with neurological signs and assessment of extremely and very low birth weight infants and to discover early any abnormal findings of diseases such as neonatal asphyxia, intraventricular haemorrhage, periventricular leukomalacia, neonatal seizures and hydrocephalus. PMID- 11905010 TI - [Evaluation of the developing human visual system using flash-visual evoked potential]. AB - Flash visual evoked potential (Flash-VEP) is easily recorded in preterm infants. However, its clinical application has not been established due to its great variability in response. Our longitudinal studies on the two components of the N1 wave facilitated peak definition and established normal ranges that are clinically valuable. The N1a (early component of the N1) peak latency decreases at about 4.6 msec/week between 30 and 40 weeks postmenstrual age. A flash-VEP study in the preterm period enables us to observe the neuronal development in the human visual system that normally proceeds in utero. Flash-VEP analyses on preterm infants demonstrated that the decrease in the N1a peak latency reflects the progress of myelination in the visual pathway according to the developmental program irrespective of preterm birth. The developmental changes of the N1 wave configuration reflect the maturation of the neuronal networks in the visual cortex, which is accelerated by extrauterine visual experience. Using improved methodology and peak denomination that we proposed, flash-VEP can be applied to preterm infants safely, and should provide us with neuro-developmental information of the human cerebrum. PMID- 11905011 TI - [Approach by neuroimaging with power flow Doppler imaging]. AB - Doppler sonography is convenient and useful for evaluating intracranial lesions and hemodynamics, especially in the neonate. Recently we used power flow Doppler imaging (PF) to show vessels with a low flow velocity and a small caliber. The pulsed Doppler system (PD) in combination with PF can selectively visualize small vessels and slow blood flow, such as the steady flow conditions of the lenticulostriate artery (LSA). As PF can reveal the vessel crossing at a right angle to the ultrasonic beam, the bilateral middle cerebral arteries can be observed through the anterior fontanel under the same conditions. We can also evaluate the laterality of intracranial hemodynamics using PF. Four-dimensional PF images, which are reconstructed from the several three-dimensional reconstruction images during one cardiac beat, can show the pulsation of intracranial arteries. This technique may provide a new quantitative and qualitative method for intracranial circulation. PMID- 11905012 TI - [The effectiveness of early intervention for very-low-birth-weight infants]. AB - We reviewed the concept and history of early intervention (EI). We summarized the results of three randomized controlled trials for high-risk young children: Abecedarian, CARE, and the Infant Health Developmental Program in the USA. All of those interventions showed positive effects on child 1Q during the first 3 years of life. A randomized EI trial for very-low-birth-weight (VLBW) infants at 8 institutions in Japan also showed enhancement of some aspects of development, including behavioral problems, circadian rhythm, and speech. The present problems of EI for VLBW infants in Japan include lack of financial support by the government, lack of evidence to determine the onset, intervals and duration of the service, and difficulties in objective evaluation of its short/long term efficacy. To enhance the maximum development of VLBW infants, pediatric neurologists should advocate the importance of EI to the government. PMID- 11905013 TI - [ADHD and remedical intervention]. PMID- 11905014 TI - [Proposed immunization program for febrile seizures(discussion)]. PMID- 11905015 TI - [Study of nine severely disabled children with hypouricemia]. PMID- 11905016 TI - [Childhood epilepsy in Okayama Prefecture, Japan--a neuroepidemiological study]. AB - Epidemiological surveys on epilepsy have been carried out repeatedly in Japan. However, at present, a population-based survey according to the International Classification for Epileptic Syndromes is considered to be especially important. We therefore carried out a population-based survey on children with epilepsy under thirteen years of age, who resided in Okayama Prefecture on December 31, 1999 as the prevalence day. The population of children in Okayama Prefecture under thirteen years of age on the prevalence day was 250,997. The lists of children with epilepsy were collected from the medical records of 45 hospitals, 36 clinics and 3 institutions in and around Okayama Prefecture. 1) 2,222 cases with active epilepsy were identified. The prevalence rate was 8.9 per 1,000. 2) This prevalence rate was almost equal to the previous result (8.2 per 1,000) given by another survey undertaken in 1975 for children under ten years of age in Okayama Prefecture. 3) If the cases with single seizure and/or the cases with seizures induced by fever are excluded, the prevalence rate decreased to 5.5 per 1,000. 4) 2,026 (91.2%) of the 2,222 cases were classified into three major categories of the International Classification of Epileptic Syndromes. They consisted of 1,557 cases (76.8%) with localization-related epilepsy, 449 cases (22.2%) with generalized epilepsy and 20 cases (1.0%) with undetermined epilepsy. 5) 303 (15.0%) of the 2,026 cases were classified into the epileptic syndromes. However, the majority of other cases consisted of nonspecific types of epilepsy, and could not be classified in detail. 6) The guideline for informed consent in epidemiological study published in 2000 by the Committee of the Japanese Ministry of Public Welfare will be useful for neuroepidemiological studies in the future. PMID- 11905017 TI - [Neurosurgery in the 21st century--today's status and future prospects of the Japanese Society of Spinal Surgery]. PMID- 11905018 TI - [Neurosurgery in the 21st century--neurotraumatology]. PMID- 11905019 TI - [Neuroendoscopic surgery--present and future]. PMID- 11905020 TI - [Intraoperative monitoring of hearing function in the removal of cerebellopontine angle tumor: auditory brainstem response and cochlear nerve compound action potential]. AB - Intraoperative hearing monitoring may be the best approach for hearing preservation in the removal of cerebellopontine angle (CPA) tumors. We designed an intracranial electrode for a reliable and consistent recording of the cochlear nerve compound action potential (CNAP), and compared the efficiency of the CNAP and auditory brainstem response (ABR) while monitoring the CPA tumor removal. Simultaneous intraoperative monitoring of CNAP and ABR were performed in 7 patients with CPA tumors (5 acoustic neurinomas and 2 epidermoids) undergoing tumor removal aimed at hearing preservation. We designed an intracranial electrode for CNAP monitoring made of a fine malleable insulated wire with a tiny tuft of oxidized cellulose on the tip. Postoperatively, the hearing of 6 patients was preserved. Useful hearing was preserved in 5 of those. ABR recordings were unsatisfactory because of severe artifacts during the tumor removal. It was discernible only in 2 of the 7 patients. Postoperatively, useful hearing was preserved in the 2 patients with a discernible ABR during the tumor removal. Four of 5 patients with no discernible ABR during the tumor removal preserved hearing. Before the tumor removal, all patients showed a reproducible CNAP of an amplitude 20 times larger than the amplitude of ABR without artifacts. A reliable CNAP was recorded consistently throughout the tumor removal in 5 patients, in whom useful hearing was preserved postoperatively. CNAP disappeared completely during the tumor removal in 1 patient who lost hearing postoperatively. Because of severe artifacts and poor specificity, the intraoperative ABR monitoring can not predict the postoperative hearing condition. Our newly designed intracranial electrode enables consistent CNAP recording during tumor removal. CNAP reflects the effect of surgical manipulations on hearing and predicts the postoperative hearing condition. CNAP is a more efficient form of intraoperative monitoring than ABR during CPA tumor removal. PMID- 11905021 TI - [Craniotomy side for neck clipping of the anterior communicating aneurysm via the pterional approach]. AB - The safety and reliability of neck clipping of the anterior communicating artery (Acom) aneurysm via the pterional approach was evaluated in terms of craniotomy side in 39 consecutive cases operated on by the senior surgeon from April 1991 through March 2000. These aneurysms were approached in principle via the side where the proximal A2 portion of the anterior cerebral artery was located posteriorly, for the purpose of easier identification of all five arteries involved, i.e., A1 and A2 portions of the anterior cerebral arteries of both sides and Acom. All aneurysms were clipped safely irrespective of the approach side because it was possible prior to aneurysmal dissection to prepare both A1 portions of the anterior cerebral arteries for temporary clipping, but not as far as the place where the aneurysm projects inferiorly and its fundus adheres firmly to the optic chiasm. The security of perforating arteries, however, could not be confirmed even after the completion of neck clipping in 9 cases. Clipping was impossible in the other 2 cases. In 2 of these 11 aneurysms the difficulty in clipping was not based on what side was used for craniotomy but on their large size. In the remaining 9 aneurysms, the necks of which were all situated on the posterior wall of the Acom, the craniotomy side turned out to be inappropriate when they were approached via the side where the proximal A2 portion of the anterior cerebral artery was located posteriorly. It was concluded that the craniotomy side should be selected so that the surgeon can observe directly the neck of the aneurysm. PMID- 11905022 TI - [Clinical usefulness of multi-planar reconstruction images of three-dimensional computed tomographic angiography for internal carotid artery aneurysms]. AB - The usefulness of multi-planar reconstruction (MPR) images of three-dimensional computed tomographic angiography (3D-CTA) for the diagnosis of internal carotid artery (ICA) aneurysms is described. Eleven unruptured ICA aneurysms including six cases of IC-cavernous aneurysm, two cases of IC-ophthalmic artery aneurysm, two cases of IC-posterior communicating artery aneurysm and one cases of IC anterior choroidal artery aneurysm, were examined by magnetic resonance angiography (MRA), digital subtraction angiography (DSA), 3D-CTA and its MPR images. 3D-CTA and DSA were useful to identify the aneurysmal neck in small aneurysms, but it was difficult to identify the aneurysmal neck in small aneurysms by 3D-CTA-MPR images. DSA and MRA were not useful for identifying the aneurysmal neck in aneurysms more than 10 mm in diameter, as a precise viewing of the neck could not be found due to their large size. For large aneurysms, neither was 3D-CTA useful for identifying the aneurysmal neck when their large size and surrounding bony structures overlapped the aneurysmal neck. On the other hand, 3D CTA-MPR was very useful for identifying the aneurysmal neck without overlapping by surrounding bony structures. 3D-CTA-MPR images clearly visualized the calcification of the wall. 3D-CTA-MPR images are obtained from 3D-CTA source images without any additional stress to the patients, and they are more useful for the diagnosis as well as demonstration of the aneurysmal neck particularly in more than large aneurysms. PMID- 11905023 TI - [Percutaneous transluminal angioplasty in fibromuscular dysplasia of the internal carotid artery: case report]. AB - We report a case of symptomatic cervical carotid artery stenosis associated with fibromuscular dysplasia (FMD) successfully treated by percutaneous transluminal angioplasty (PTA). A 49-year-old female presented with repeated ischemic attacks of right hemiparesis and aphasia. Digital subtraction angiography revealed a string of beads appearance of the left internal carotid artery typical of the medial type of FMD, at the level of the C2 vertebra. Similar findings were also evident in the right internal carotid artery at the level of the C1 vertebra, but without significant narrowing. MR angiogram of the left carotid artery revealed a marked flow gap, suggestive of severe stenosis. Forty days after onset PTA was indicated for symptomatic FMD of the left internal carotid artery because of the relative difficulty in approaching and repairing this highly located lesion surgically. The stenotic lesion was very easily dilated without any procedural complications. Although angiography just after PTA showed slight residual irregularity of the wall, this smoothed up with time. Follow-up MR angiography 7 years after PTA demonstrated long-term patency without clinical evidence of deterioration. Patients with the medial type of FMD in the carotid artery seem to be excellent candidates for PTA. PMID- 11905024 TI - [Combination of intravascular surgery and surgical operation for occipital subcutaneous arteriovenous fistula in a patient with neurofibromatosis type I]. AB - We presented a case of neurofibromatosis type I (NF-1) associated with an extracranial arteriovenous fistula (AVF) fed by the occipital and vertebral artery. A 20-year-old man was referred to our hospital because of an occipital subcutaneous pulsatile mass. A CT scan showed a huge subcutaneous enhanced mass. Angiography revealed that the occipital AVF was fed by bilateral occipital arteries, the left ascending pharyngeal artery, the left middle cerebral artery, and the left vertebral artery with-abundant communication with the subcutaneous veins. Endovascular treatment by using both coil and glue (Eudragid) embolization via the occipital artery successfully obliterated the AVF. Subsequently surgical operation was performed. Postoperative angiography showed the disappearance of the AVF. Combination of intravascular surgery and surgical operation should be considered as an effective treatment for NF-1 associated with AVF. PMID- 11905025 TI - [A case of a ruptured pseudoaneurysm arising from the distal part of the posterior inferior cerebellar artery]. AB - We report a rare case of a pseudoaneurysm arising from the distal part of the posterior inferior cerebellar artery (PICA) and which was associated with intraventricular hemorrhages. A 48-year-old female had sudden onset of severe headache and vomiting, and was admitted in a semicomatous state to our hospital. Initial CT scan showed ventricular hematomas resulting in acute hydrocephalus, but no obvious parenchymal or subarachnoid hemorrhage. Ventricular drainage was carried out. Subsequently, cerebral angiography was performed, but neither, an aneurysm nor other vascular abnormality was detected. 50 days after admission, MRI revealed a homogenous and well-circumscribed hypointense mass (12 mm in diameter) in the right cerebellar tonsil, suggesting the presence of hemosiderin. 60 days after the onset, the second angiography showed an aneurysmal shadow arising from the distal part of the PICA. An aneurysmal clipping was performed via the suboccipital approach. A thrombosed aneurysm was identified in the right tonsil. At the time of the operation, we confirmed that the part of the PICA around the aneurysm was intact with neither branching nor discoloration. Histological examination revealed that elastic fiber was not identified in the aneurysmal wall, and the aneurysm was diagnosed as a pseudoaneurysm. Histories of trauma or infection, which may have induced the cerebral pseudoaneurysm, were not obtained by interview. PMID- 11905026 TI - [Treatment for the ruptured bilateral vertebral dissecting aneurysms]. AB - We report a case of a bilateral vertebral dissecting aneurysm associated with subarachnoid hemorrhage. Proximal ligation of the vertebral artery on the ruptured side combined with wrapping of the contralateral dissection failed to prevent fatal rebleeding. Since enlargement of the contralateral dissection was observed by postoperative angiography, rupture of the growing contralateral dissecting aneurysm may have caused rebleeding. Hemodynamic changes following the occlusion of one vertebral artery might have led to enlargement and subsequent rupture of the contralateral dissection. Direct wrapping was unable to prevent enlargement of the dissection, so radical surgery including bilateral vertebral artery occlusion combined with vascular reconstruction may be the treatment of choice for this type of lesion. PMID- 11905027 TI - [The usefulness of 3D-CTA for the diagnosis of a ruptured aneurysm at the origin of the duplicated middle cerebral artery: case report]. AB - Duplication of the middle cerebral artery is known as a rare anomalous vessel arising from the internal carotid artery and an aneurysm at the origin of the duplicated middle cerebral artery is very rare. We presented a case of ruptured aneurysm at the origin of the duplicated middle cerebral artery and discussed the usefulness of 3D-CTA (three-dimensional computed tomographic angiography) for its diagnosis. A 34-year-old female suffered from severe headache and was admitted to our hospital. CT scan revealed diffuse subarachnoid hemorrhage and angiography revealed duplication of the right middle cerebral artery and dilatation at its origin. We could not identify it as an aneurysm by angiography, so we performed 3D-CTA. 3D-CTA was able to demonstrate clearly the aneurysm at the origin of the duplicated middle cerebral artery and we performed neck clipping of the ruptured aneurysm. To our knowledge, previously there have been only 14 cases which reported such an aneurysm at the origin of a duplicated middle cerebral artery. We reviewed the 15 cases including ours and found that, in 4 cases, the aneurysm could not be detected by the initial angiography. We suspected that most of these aneurysms were small, so the detection of the aneurysms by angiography was difficult. We conclude that 3D-CTA is useful for diagnosing aneurysms at the origin of the duplicated middle cerebral artery even when thy can't be detected by angiography. PMID- 11905028 TI - [Tuberculosis in the patients undergoing haemodialysis in Japan, 1996]. AB - We conducted a questionnaire survey on patients undergoing haemodialysis about the present situation of tuberculous incidence. They are immunocompromised hosts and are said to be at high risk of developing tuberculosis in many reports. (1) DESIGN Of the 167,192 patients on haemodialysis registered on December 31, 1996 in Japan, 71,411 patients were available for the questionnaire survey. Of the 2,893 hospitals used as the study subjects, 1,108 hospitals gave satisfactory replies. Of them, 141 hospitals reported that they had patients with tuberculosis in 1996, and 79 cases were collected by the detailed survey on tuberculosis patients conducted later. They included 45 male cases, 34 female cases for tuberculosis of all forms, 28 male cases, 15 female cases for pulmonary tuberculosis (PTB), 13 male cases, 4 female cases for tuberculosis bacilli positive pulmonary tuberculosis (TB positive PTB), and 17 male cases, 19 female cases for extrapulmonary tuberculosis. (2) RESULTS: In tuberculosis of all forms, the number of observed patients (O) against the number of patients expected (E) was calculated, and the standardized patients ratio (O/E ratio) was computed. It was 1.55 for male, 2.79 for female and 1.99 for total. The incidence of tuberculosis haemodialysis patients was significantly higher compared with the general population (p < 0.01). As to PTB, the O/E ratio was 1.01 for male, 1.40 for female and 1.16 for total; the incidence of PTB was not significantly higher compared with the general population. With TB positive PTB, the O/E ratio was 0.96 for male, 0.80 for female and 0.97 for total, and no significant difference was found. As for extrapulmonary tuberculosis, the O/E ratio was 13.45 for male, 13.07 for female and 12.97 for total; the incidence of extrapulmonary tuberculosis in haemodialysis patients was significantly higher (p < 0.01), but it was lower than these reported in the past literature. The seventy nine cases consisted of 52 primary treatment cases, 23 retreatment cases, and 4 unknown cases. Out of 79 cases, 36 cases developed tuberculosis almost at the same time or within 1 year after undergoing haemodialysis, and thereafter it decreased gradually. Underlying diseases for haemodialysis were mainly glomerulonephritis and diabetic nephropathy. There were many patients who failed to notify to the public health centers after the diagnosis of tuberculosis was made, and it is needed to improve such a situation in the future. The prognosis of tuberculosis undergoing haemodialysis was poor. Three out of 43 patients with PTB and 2 out of 13 tuberculosis pleurisy cases died. (3) CONCLUSION: The risk of developing PTB in patients undergoing haemodialysis was not high compared with the general population, however, the risk was much higher for extrapulmonary tuberculosis. Moreover, the treatment outcome was not satisfactory in patients with PTB and pleurisy. As patients undergoing haemodialysis have the factors which suppress the cell-mediated immunity, it is required to restudy the measures to prevent development of tuberculosis, management and treatment in the future. PMID- 11905029 TI - [Interpretive compatibility of antimycobacterial susceptibility for Mycobacterium tuberculosis determined by proportion test method on egg-based Ogawa media and broth microdilution test, BrothMIC MTB]. AB - The antimycobacterial susceptibility test method newly proposed by the Japanese Society for Tuberculosis, a proportion method on egg-based Ogawa media, was evaluated in comparison with microdilution test for Mycobacterium tuberculosis complex, BrothMIC MTB-1 (Kyokuto Pharmaceutical Inc., Tokyo). In the evaluation, five antimicrobial agents, streptomycin, ethambutol, kanamycin, isoniazid and rifampicin were included. Through repeated testings of the three reference strains against five antimicrobial agents, both test methods were found to be highly precise. All the minimum inhibitory concentrations (MICs) determined by BrothMIC MTB fell within 3 log2 dilutions, however a total of 11 MICs resulted in indeterminate(I) interpretations. Whereas, all the test results by a proportion method on Ogawa media were comparable to the expected interpretations. However, three of 48 testings resulted in undeterminable interpretations due to insufficient growth on the growth control media. A total of 127 clinical isolates of M. tuberculosis complex were tested by both methods, and 89 to 90% of the test results were comparable with each other in category interpretations. However, 7.1 to 9.4% of MICs determined by BrothMIC MTB resulted in indeterminate(I), and 0.8 to 3.1% of discrepant interpretations were observed. In conclusion, both test methods were highly precise and comparable in determining antimycobacterial susceptibility for M. tuberculosis complex. Several advantages and disadvantages in each test method were discussed. PMID- 11905030 TI - [A case of miliary tuberculosis complicated with SIADH, brain tuberculoma, and tuberculous meningitis]. AB - A 46-year-old man complained fever, headache, and vertigo after he was given steroid for sudden deafness. He was diagnosed as miliary tuberculosis by his chest CT findings. After admission, 4 anti-tuberculous drugs (INH, RFP, SM, and PZA) were prescribed but his laboratory findings showed SIADH, which was difficult to treat, and steroid was readministered. Brain MRI, examined 2 months after admission, showed brain tuberculomas, and examination of cerebrospinal fluid revealed a diagnosis of tuberculous meningitis. Three months later, meningitis deteriorated transiently, however symptoms and findings improved by increasing steroid. Later, miliary tuberculosis and SIADH were cured, however, some tuberculomas grew larger gradually on brain MRI, and spinal MRI showed tuberculomas in the spinal cord. LVFX, high concentration in CSF, was added. At present (2 yrs after beginning the therapy), lesions in the brain and spinal cord improved but remain with the sequelae. PMID- 11905031 TI - [A case of disseminated tuberculosis requiring extended period for the identification of Mycobacterium tuberculosis on culture]. AB - A 80-year-old male visited an outpatient department of a nearby hospital complaining of fever, cough, and poor appetite on June 2000. The patient was diagnosed as bacterial pneumonia and was treated with antibiotics although specific cause could not be identified. After one month, he was hospitalized due to lack of improvement. After admission, acid-fast bacilli (AFB) was found from the bronchial washing. The patient was then transferred to our hospital. Upon admission, sputum smear examination was positive for AFB and MTB was confirmed by PCR. Therapy was initiated with INH 300 mg, RFP 450 mg, EB 1000 mg, and PZA 1000 mg, orally daily. However, on the day following the admission, he became unconscious. Brain MRI showed several small granulomas on the cortex of the bilateral anterior and temporal brain. Although AFB was not detected from the cerebrospinal fluid, tuberculous meningitis was suspected and steroid was given. Nine days after admission, the patient died due to tuberculous meningitis. The isolation of MTB had been attempted on Ogawa culture medium using patient's sputum and liquor, and it took 14 weeks to find colony growth both from sputum and liquor. In the autopsy, numerous granulomas were detected in his lung, liver, kidney, and pancreas. These findings indicate that disseminated growth of MTB occurred in vivo in spite of very slow growth of MTB in vitro. PMID- 11905032 TI - [CT diagnosis of pulmonary infectious diseases]. AB - Generally, the patient's inflammatory reaction is obvious, and the characteristic infiltrative shadow as pneumonia in the chest radiograph is recognized, no more imaging study is needed. However, when (1) radiographic finding is uncommon as a pneumonia; (2) antimicrobial drug treatment is ineffective; or (3) stenosis of central bronchi causing pneumonia by lung cancer and so on is suspicious, more precise imaging information of computed tomography (CT) is necessary. Recently, high resolution CT of lung disease as a new diagnostic method has been introduced, and diagnostic efficacy of CT has markedly advanced. Although the high resolution CT diagnosis based on the minute anatomy of the lung is also applied for the pulmonary infectious disease, their analysis and arrangement is not yet established since a large number of pathogens and various images exist. In reading CT films, it is necessary to consider patient's immune status. Infectiosity and size of the pathogen must be also considered. In this manuscript, CT imaging characteristics of lung infectious diseases are clarified by taking these points into account. PMID- 11905033 TI - [Regulation of Th1 and Th2 immune responses by IL-18]. AB - IL-18, which requires cleavage with caspase-1 to become active, was originally discovered as a factor that enhances IFN-gamma production from Th1 cells in the presence of anti-CD3 or anti-TcR Ab. However, it was later shown that IL-12 and IL-18 without TcR engagement can induce IFN-gamma in Th1 cells and nonpolarized T cells. Additional TcR engagement has no effect on this IFN-gamma response. Furthermore, a combination of IL-12 and IL-18 acts on B cells, NK cells, macrophages and dendritic cells to produce IFN-gamma. In contrast, IL-18 without help from IL-12 induces Th2 cytokines in T cells and NK cells. Moreover, IL-18 directly stimulates basophils and mast cells to produce Th2 cytokines and histamine independently of IgE. Most surprisingly, IL-18 causes high-level IgE production when administered to normal mice by causing CD4+ T cells to produce IL 4 and to express CD 40 ligand. We established skin-specific caspase-1 transgenic mice with elevated levels of IL-18 in their sera. We found high serum level of IgE, which is entirely dependent on stat 6 in these transgenic mice. These results indicate that caspase-1/IL-18 may be critically involved in regulation of IgE production in vivo, providing a potential therapeutic target for allergic disorders. PMID- 11905034 TI - [Current and future state of chemoradiotherapy for head and neck cancer]. AB - Radiation therapy was the conventional treatment for locally advanced, nonresectable head and neck cancer. However, therapeutic results were poor with this treatment modality, and chemoradiotherapy has been used in an effort to improve therapeutic results. Improved local-regional control and disease-free or overall survival have been shown in several randomized trials using a concurrent or alternative approach. Induction chemotherapy (neoadjuvant chemotherapy), however, has not been shown to improve local-regional control or survival. Induction chemotherapy followed by definitive radiotherapy may be useful in the selection of patients who are likely to benefit from non-surgical organ preservation treatment schemes. Further clinical trials are needed to clarify the most suitable combination of chemotherapy and radiation. Intraarterial chemotherapy combined with radiation therapy for head and neck cancer has been attempted for many years. However, the indications, clinical significance, and selection of suitable anti-cancer drugs remain unclarified. The modern superselective intraarterial approach should be re-evaluated. Many head and neck cancers have been found to overexpress the receptor to epidermal growth factor (EGFR). Antibodies such as IMC-C225 that specifically target EGF receptors with radiotherapy and/or chemotherapy may prove to be valuable contributors to the treatment of advanced head and neck cancer. PMID- 11905035 TI - [MR imaging of small bowel with water administration]. AB - We performed MR imaging of the small bowel (MRSB) in 20 patients using water as an oral contrast agent, to improve the demonstrability of pathologic conditions without a large amount of intestinal fluid. Bowel lumen and folds were clearly visualized: duodenum in 13(65%), jejunal loops in 14 (70%), ileal loops in 15(75%), and ileocecum in 8 (40%) cases. Furthermore, conventional enteroclysis was performed in 16 of 20 patients, and the MRSB findings were comparable with those of conventional enteroclysis. If conventional enteroclysis is used as the gold standard, MRSB visualized luminal stenosis in 11 of 13(84.6%), displacement or extrinsic compression in 4 of 5(80%), polypoid lesion in 3 of 4(75%), and fistula formation in one of one cases. None of four ulcerative lesions could be visualized by MRSB. Our MRSB technique is a noninvasive, easy method that does not require a long time. Accordingly, MRSB can be used in addition to the conventional MR sequence. MRSB has potential usefulness for evaluating small bowel disease without radiation exposure. PMID- 11905036 TI - [Report on proton therapy according to good clinical practice at Hyogo Ion Beam Medical Center]. AB - The Hyogo Ion Beam Medical Center(HIBMC) is a hospital-based charged particle treatment facility. Having two treatment ion beams(proton and carbon) and five treatment rooms, it is a pioneer among particle institutes worldwide. In May 2001, proton therapy was started as a clinical study for patients with localized cancer originating in the head and neck, lung, liver, and prostate. The aim of this study was to investigate the safety, effectiveness, and stability of the treatment units and systems based on the evaluation of acute toxicity, tumor response, and working ratio of the machine, respectively. Six patients, including liver cancer in three, prostate cancer in two, and lung cancer in one, were treated. There was no cessation of therapy owing to machine malfunction. Full courses of proton therapy consisting of 154 portals in all six patients were given exactly as scheduled. None of the patients experienced severe acute reactions of more than grade 3 according to NCI-CTC criteria. Tumor response one month post-treatment was evaluable in five of the six patients, and was CR in 1 (prostate cancer), PR in 2 (lung cancer: 1, liver cancer: 1), and NC in 2(liver cancer: 2). These results indicate that our treatment units and systems are safe and reliable enough for proton irradiation to be used for several malignant tumors localized in the body. PMID- 11905037 TI - [Trial production of water substitute phantom considering the effect of light]. AB - We have produced a novel water substitute phantom suitable for film dosimetry, while retaining the radiological property of a conventional water substitute phantom. The novel phantom excludes the effect of light, which is known to affect the accuracy of results on conventional phantoms. The effect of light was eliminated by appropriately adjusting the quantum of the carbon black to that of a conventional phantom material. Through comparison of the novel phantom with a conventional phantom it was shown that the absorbed dose determined by conventional phantom was 15% higher for 10 MV X-rays and 18% higher for an 18 MeV electron beam, attributable to the contamination of Cerenkov light. Although the net optical density of the film increased with time owing to the optical permeability of the phantom, that of the novel phantom did not vary with time. The novel phantom was therefore shown to be unaffected by such local light and by the optical transmission of the phantom. PMID- 11905038 TI - Malformation effects in ddY mice irradiated at two stages in the preimplantation period. PMID- 11905039 TI - [Application of immunohistochemistry for forensic pathological diagnosis: finding of human brain in forensic autopsy]. AB - We researched the application of immunohistochemistry for the purpose of establishing forensic pathological diagnoses. In the present study, we examined the induction and expression of heat shock protein (HSP), oxygen regulated protein (ORP), inducible nitric oxide synthase (iNOS), excitatory amino acid transporter 2 (EAAT2) and apolipoprotein E (apo E) in the human brain using forensic autopsy cases as our subjects. Hypoxic/ischemic brain damage. In cases of longer survival and with a history of hypoxic attacks, the proteins HSP and ORP were found in the parieto-occipital lobe and hippocampus. And we are able to observe a weak stain for EAAT2 in almost all asphyxia deaths. Traumatic brain injury (TBI). In traumatic brain injury (TBI), the prolonged induction of iNOS was demonstrated in the neutrophils, microglia/macrophage, and vascular smooth muscle cells in the traumatized brain. Apo E was identified with neurons in the traumatized cortical hemisphere from only a two-hour survival case to long survival cases. To the contrary, there was no positive apo E staining in the contralateral cortical hemisphere at all. In one one-hour survival case, a weak stain for EAAT2 was observed, but intensive expression of EAAT2 was observed from brief to one-day survival cases. Sudden infant death (SID). Numerous ferritin positive cells were observed in the brain in the cases of pneumonia or myocarditis that we examined. To the contrary, the numbers of ferritin-positive cells were obviously decreased in the cases of sudden infant death syndrome (SIDS). The transferrin-positive cells were in an inverse proportion to the ferritin positive cells in each SIDS case. Also, numerous ORP-150 positive cells were observed in the brain in cases of pneumonia and the SIDS group. In forensic practice, immunohistochemical investigation of these proteins can be a great value for diagnosing not only the cause of death but also the pathophysiological changes and the victims past history. PMID- 11905040 TI - [New development of hair examination! The searching of hair for the terminal residual substance of pleasant sensation]. AB - Formerly, hair had been analyzed mainly in the field of individual identification. Otherwise, it is known that many substances are accumulated in hair such as metals like arsenic, mercury, alkaloids like caffeine, theophylline, drugs like opiate, stimulants, other drugs and poisons. So, by qualitative and quantitative analysis of such deposits in hair. It is possible to presume one's life manner during recent several months. This is the new development of hair analysis and it is expected that it may be wide of application. The main theme of this paper is hedonics. Nobody dislike hedonic sensation. So, it can be said everybody being living always coveting hedonic sensation. There are many hedonic sensers all over the body, and hedonic sensation transmitted by A10 nerve, so A10 nerve is called the hedonic nerve. Neurotransmitter of this A10 nerve is Dopamine. Dopamine is the endogenous hedonic material. Exogeneous hedonic one is stimulant like Methamphetamine. In this paper, relation between Dopamine, Methamphetamine and Tyramine, thought to be the terminal product from two formers is examined in the aspect of hair analysis. Catecholamines including Dopamine are made of Tyrosine. They have a Benzene-ring. Compounds with a Benzene-ring made of Tyrosine are relatively rare, but all of them have very strong action. Tyramine is made of Tyrosine, too. So the structure of Tyramine is essentially resemble to Dopamine, and the skeletal structure of Methamphetamine is identical to Dopamine, so the action is identical, too. Normally, excessive release of Dopamine in brain is controlled by the negative feedback of GABA system. But extrinsic stimulus as Methamphetamine put balance of the homeostasis between Dopamine system and GABA system in to disorder, and long term administration of Methamphetamine makes long remaining degeneration of synapse of Dopamine nerve. So, abnormal state of excess releasing of Dopamine happens easily. And the morbid state named stimulant psychosis occur. The volume of Tyramine in hair changes correlatively with the volume of released Dopamine in brain. Therefore it is suggested that quantitative analysis of Tyramine in hair enable to estimate the changes of Dopamine released in brain. PMID- 11905041 TI - [Cardiac lesions in methamphetamine abusers]. AB - Various cardiac lesions such as hypertrophy, disarray and fibrosis similar to HCM, were often found in the heart of methamphetamine (MA) abusers. Myolysis, eosinophilic changes, contraction band necrosis and small round cell infiltration were also observed. Male ddy mouse were administered MA 1 mg/kg subcutaneously every day for 4 weeks. Their hearts revealed many cardiac changes such as hypertrophy, myolysis, contraction band necrosis, disarrangement of myofibers, saw-like cytoplasm, side-to-side connection of cardiac cells and vascuolative degeneration microscopically, and crysterosis of mitochondria, enlargement of sarcoplasmic reticulum and hypercontraction electronmicroscopically. These changes are thought to be similar to that of MA abusers, so it is certified that MA has toxic effect on the heart. Moreover, these changes could not be found when beta-blocker or calcium antagonist was premedicated. To elucidate the mechanisms of MA cardiac toxicity, we have designed some experiments. When MA (15 mg or 20 mg/kg) was administered on rats, cardiac lipid peroxidates, as a marker of free radical, increased rapidly. When rats were feeded for 7 weeks with Vitamin E deficient diet, 10 mg/kg MA administration was enough to increase lipid peroxidates. Simultaneous ECG observation revealed various arrhythmia such as VPB, A-V block and intraventricular conduction delay. In the investigation of contractile protein, although we could not find differences in the isozyme pattern of myosin heavy chain between MA groups (1 mg/kg for 8 and 12 weeks) and control group, Mg2+ ATPase activity of myocardial actomyosin at 0.1 microM Ca2+ increased significantly in 12 weeks MA group. We also found MA induced cardiac toxicity in cultured myocytes. Primary cultured adult rat myocytes were exposed to MA (1 x 10(-5) M and 1 x 10(-3) M) for 1 to 24 h in the presence and absence of 1 x 10(-5) M propranolol. After 24-h MA treatment, cellular granulation, swelling and hypercontraction and release of CPK were observed both with and without propranolol treatment. These findings suggest that MA may exert direct toxic effects on the heart. PMID- 11905042 TI - [The false positive reaction of the Triage panel drug-of-abuse by herbal drugs ma huang (Ephedra sinica (Ephedraceae))]. AB - We investigated false-positive reactions obtained from a drug screening test using a Triage panel. We detected 2 cases giving false-positive reaction for AMP (amphetamine, methamphetamine) during the screening of 187 normal subjects. Subsequent follow up testing by high-performance liquid chromatography (HPLC), showed both to be false-positive reactions. As both cases have a history of ingesting the herbal drug, Ma-huang (Ephedra sinica (Ephedraceae)), containing ephedrine, we examined the relationship between false-positive reactions on Triage and Ma-huang. All urine samples collected from 7 healthy volunteers following administration of Ma-huang indicated AMP positive on Triage. Also a high ratio of AMP positives was observed in the patients who were administered Ma huang-containing drugs at the hospital. However, none of them were identified as true-positives by HPLC or gas chromatography mass spectrometry (GC/MS) analysis. The extract of Ma-huang contained in herbal drugs, which otherwise contain neither amphetamine nor its derivatives, gives (AMP) positive indications on Triage. We speculate that unidentified components of Ma-huang cause the false positive reactions. We suggest that follow-up tests by GC/MS or HPLC are needed wherever a positive result is obtained from a screening test by Triage. Furthermore, it will be established to continue collecting information on prescribed and non-prescribed drugs. PMID- 11905043 TI - [Radiographic examination of skeletalized remains of an abandoned infant: a case report with special reference to odontological findings]. AB - A homeless couple confessed that they had buried their baby, who died at the age of three months, in the graveyard two years earlier. The recovered infantile remains had been already completely skeletonized and dismembered. However, the maxilla and mandibula of the average matured infants, the age of the body was estimated to be 1 to 2.5 months old on the basis of the degree of calcification of deciduous teeth. PMID- 11905044 TI - [An autopsy case of an infant born with a caul who died due to asphyxia after live-birth]. AB - We reported an autopsy case of a child born with a caul. The infant was a female and seemed to be full-term. No remarkable pathological findings for disease or anomalies were observed. The floating test of lungs was partially positive. And on the gastro-intestinal floating test, only stomach floated. In histological examination, the alveoli in the lungs were partially slightly open. We thought that those were due to the cardio-pulmonary resuscitation. Pulmonary surfactant was positive immunohistochemically. And the stable microbubble test on the liquid in her stomach was very strong. Even if a child is born alive, he or she will soon die due to asphyxia but for the production of surfactant in the lungs. We can evaluate more precisely the potential respiratory ability of a neonate died due to asphyxia by means of stable microbubble test on newborn gastric contents and immunohistochemical staining of surfactant in lungs. PMID- 11905045 TI - [A judgement in litigation over return of pathological samples]. AB - The son of a deceased patient filed a law suit against a medical school hospital for return of all the pathological samples (paraffin blocks and preparations) made from part of the patient's corpse. The reason cited is that the samples included at least one type about which he had not given consent. The Tokyo distinct court passed a judgement, rejecting the defendant's claim that the consent was meant to be all-inclusive, and ordered the defendant to return all the samples still in their possession to the litigant. This judgement made the legal meaning clear as to the permission of a decreased person's family, both from the standpoint of public law and from that of private law. The judgement made the first impression on permitting their ownership. PMID- 11905046 TI - [Formation and removal of reactive oxygen species, lipid peroxides and free radicals, and their biological effects]. AB - It is well known that biomembranes and subcellular organelles are susceptible to lipid peroxidation. There is a steadily increasing body of evidence indicating that lipid peroxidation is involved in basic deteriorative mechanisms, e.g., membrane damage, enzyme damage, and nucleic acid mutagenicity. The formation of lipid peroxides can be induced by enzymatic or nonenzymatic peroxidation in the presence of oxygen. The mechanisms of formation and removal of reactive oxygen species, lipid peroxides, and free radicals in biological systems are briefly reviewed. In recent years, there has been renewed interest in the role played by lipid peroxidation in many disease states. Xanthine oxidase has been shown to generate reactive oxygen species, superoxide (O2-.), and hydrogen peroxide (H2O2) that are involved in the peroxidative damage to cells that occurs in ischemia reperfusion injury. During ischemia, this enzyme is induced from xanthine dehydrogenase. We have shown that peroxynitrite (a reactive nitrogen species) has the potential to convert xanthine dehydrogenase to oxidase. The following biological effects of lipid peroxidation were found: a) the lipid peroxidation induced by ascorbic acid and Fe2+ affects the membrane transport in the kidney cortex and the cyclooxygenase activity in the kidney medulla, and b) the hydroperoxy adducts of linoleic acid and eicosapentaenoic acid inhibit the cyclooxygenase activity in platelets. The balance between the formation and removal of lipid peroxides determines the peroxide level in cells. This balance can be disturbed if cellular defenses are decreased or if there is a significant increase in peroxidative reactions. Once lipid peroxidation is initiated, the reactive intermediate formed induces cell damage. PMID- 11905047 TI - [From new molecular science to new supramolecular science with macrocyclic polyamines]. AB - Exploitation of saturated macrocyclic polyamines has led to the discovery of numerous novel functions and new molecules such as 1) unique proton sponge properties; 2) uptake of biological polyanions; 3) peptide-like metal uptake properties; 4) stabilization of unusual oxidation states of metal ions (e.g., CuIII, NiIII); 5) novel uptake and activation of O2 by new NiII-macrocyclic complexes; 6) a new synthetic pathway to functionalize macrocyclic polyamines; 7) the first gold(III) complex that is a new candidate for gold-plating agents; 8) intrinsic zinc(II) properties pertinent to zinc enzymes; 9) selective recognition of thymine by ZnII complexes; and 10) new cage supermolecules. These newly discovered molecules and properties have opened up a new field of supramolecular science. PMID- 11905048 TI - [Development of solubility screening methods in drug discovery]. AB - We developed two methods for solubility screening of drug candidates in drug discovery. The first is a solution-precipitation (SP) method, in which the sample solutions are prepared by adding the drug solution in dimethylsulfoxide (DMSO) to buffers followed by filtering off the precipitate using 96-well filterplate. The second is a powder-dissolution (PD) method, in which the solid samples are dissolved to the buffer in the HPLC vial equipped with the filter membrane in the HPLC autosampler. An HPLC equipped with a photodiode array detector is used to measure the concentration of the sample solutions in both methods. The SP method was used for high throughput screening the solvating process of the candidates in aqueous solutions with lower sample consumption, and the PD method was used for screening both inter-molecular interaction in solid state and solvation in aqueous solution with more sample amount than that of SP method. Therefore, the solubility screening from early to final stage of lead optimization process would be successfully accomplished by using both methods complementarily. PMID- 11905049 TI - [Simultaneous determination of propanil, carbaryl and 3,4-dichloroaniline in human serum by HPLC with UV detector following solid phase extraction]. AB - In case of poisoning by herbicide compounded with Propanil (DCPA) and Carbaryl (NAC), we attempted simultaneous solid-phase extractions of DCPA, NAC, and 3,4 dichloroaniline (DCA), a metabolite of DCPA, from the patient's serum, and quantitative analytical method using HPLC-UV detection. With this HPLC method, the quantitative detection limits in the serum are 0.005 microgram/ml for DCPA and DCA and 0.001 microgram/ml for NAC, and the UV spectra of all three compounds could easily be obtained using a diode-array detection limit of 0.05 microgram/ml. When the three compounds were added to serum at concentrations ranging from 0.1-10.0 micrograms/ml, the recovery rates were satisfactory at between 91.1% and 101.9%. On analysis of the serum of patient who had ingested Kusanon A Emulsion, the ingested substance apparently caused an increase in the DCA concentration, which led to the appearance of methemoglobinemia. The possibility that the DCA concentration might be used for prognostic purposes was suggested. PMID- 11905050 TI - [Quality evaluation of essential oils]. AB - Essential oils on the market were analyzed using GC-MS and the main ingredients of each essential oil were quantified. Analysis of the essential oil of Lavandula officinalis (lavender oil) showed that each sample had a different ratio of the contents of main ingredients, such as linalool, linalyl acetate, and camphor. In addition, some commercial lavender oils were analyzed by GC-MS for comparison with the Lavandula flagrans (lavandin oil) and the reference standard. As a result of this analysis, although the components of almost all commercial lavender oils were approximately the same as those of the reference standard, there were a few products that contained more than 0.5% of the amount of camphor in lavandin oil. This suggests that some lavender oil samples are mixed with lavandin oil to lower the price. Commercial essential oils of Melaleuca alternifolia (teatree oil) and Mentha piperita (peppermint oil) were also analyzed by GC-MS. Each of the peppermint oil samples had a different ratio in the content of its main ingredient. With respect to teatree oils, the amount of terpinens in each sample differed. These results led to concern about the efficacy of essential oils. For achieve the expected efficacy of essential oils, correct information on their ingredients should be available and quality control using instrumental analysis should be introduced. PMID- 11905051 TI - [Magnetoencephalographic responses to odor and non-odor by fast Fourier transformation analysis in humans]. AB - Magnetoencephalographic (MEG) responses to odor (amyl-acetate) and non-odor stimuli for 1 second were recorded in 9 healthy volunteers (right handed) with a dual 37-channel SQUID (Magnes, Bti Co.) and evaluated by fast Fourier transformation analysis, with the following results: 1. On MEG analysis, the spectral density increase in the left mid-central region at a frequency of 7 Hz was significantly greater in response to odor than in response to non-odor stimuli. This greater increase is apparently related to the presence of the odor perception mechanism in the orbital frontal area, a major center of the olfactory system. 2. Both increased and decreased spectral density areas at a frequency of 8 Hz were observed over the right hemisphere when no stimuli was compared with non-odor and no stimulus compared with odor. These changes may reflect a high level of vigilance caused by stimulation. 3. When no stimulus was compared with non-odor stimulation, a significant spectral density increase at a frequency of 11 Hz was noted. Similar trends were observed at frequencies of 11 and 12 Hz when no stimulus was compared with odor. These findings indicated increased attention in response to random presentation of odor and non-odor. 4. Significant differences at frequencies from 14 to 24 Hz were noted in the contralateral hemisphere when no stimulus was compared with odor stimuli. MEG spectral densities at 21 and 22 Hz were also noted in the contralateral hemisphere when no stimulus was compared with non-odor stimulus. These differences apparently arise from the response of the somato-sensory cortex to non-odor stimuli and amyl acetate. Alternation of MEG spectral densities at frequencies from 14 to 17 Hz and 23 to 24 Hz in the left hemisphere was noted when no stimulus was compared with non-odor and no stimulus was compared with odor. These results appear to be related to "emotions" of pleasantness and unpleasantness evoked by non-odor and odor. PMID- 11905052 TI - [Inhibition of caspase-9 activity in cisplatin-resistant head and neck squamous cell carcinoma]. AB - We have previously reported that cisplatin induces caspase-9 activation in head and neck squamous cell carcinoma cells (HNSCCs) in vitro, and the use of a specific inhibitor of caspase-9 blocks cisplatin-induced apoptosis in HNSCCs. Our purpose here was to determine whether HNSCCs selected for resistance to cisplatin fail to exhibit caspase-9 activation in response to cisplatin. Cisplatin resistant HNSCCs (CRHNSCCs) were selected for growth in the presence of cisplatin. Following cisplatin treatment, no protelyzed caspase-9 subunits were detected in the CRHNSCCs, whereas proteolytic degradation of procaspase-9 was observed in parental cisplatin-sensitive HNSCCs (CSHNSCCs). Using a direct enzymatic assay measuring cleavage of the synthetic peptide substrate (LEHD-AFC), caspase-9 activity in cisplatin-treated CRHNSCCs was less than that in cisplatin treated CSHNSCCs. Because caspase-9 activation requires the release of mitochondorial cytochrome c (Cyt c) into the cytoplasm, we determined the level of cytoplasmic Cyt c in response to cisplatin treatment. Interestingly, following cisplatin treatment, the same extent of increase in cytoplasmic Cyt c was evident and the expression of Bcl-2 family proteins (Bcl-2 and Bcl-XL) remained unchanged in both CRHNSCCs and CSHNSCCs. These results suggest that in certain HNSCC cell types, inhibition of caspase-9 activity represents another mechanism of acquired cisplatin resistance. This inhibition mechanism may be independent of the release of Cyt c into the cytoplasm. PMID- 11905053 TI - [Tumor necrosis factor-alpha and interferon-gamma modulate in vitro expression of nitric oxide synthase in human nasal epithelial cells]. AB - BACKGROUND: Interest in the physiological, pathological and therapeutic implications of nitric oxide (NO) have grown exponentially, with human nasal cavity and paranasal sinuses considered a dominant source of NO, indicating that this molecule possesses the diversity of biological effects in the regulation of airway clearance and nonspecific cellular immunity. We previously observed differences in NO synthase (NOS) isoform constitutively expressed in nasal epithelial cells (NECs) from allergic and normal subjects. OBJECTIVES: We extended the previous work to determine whether in vitro stimulation with proinflammatory cytokines influences levels of different NOS isoform expression. METHODS: Nasal epithelial cells were sampled from the inferior turbinate in a group of 16 healthy normal controls and 11 patients with perennial allergic rhinitis against house dust (HD) mite antigen. 1 x 10(5) cells were incubated in conditioned medium for 24 hours. Human recombinant interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), or both (cytomix) were added to a final concentration of 10 ng/ml. Cells were then fixed with 4% paraformaldehyde and processed for fluorescence immunocytochemistry. Immunoreactivity for 2 NOS isoforms, inducible NOS (iNOS) and endothelial NOS (eNOS), was studied by laser scanning confocal microscopy and fluorescence intensity was assessed quantitatively. RESULTS: We observed constitutive eNOS expression in epithelial cells of all subjects. Different treatments with cytokines did not affect eNOS expression. Cytokine treatment, however, significantly augmented iNOS expression in the control group. The average increase induced with IFN-gamma, TNF-alpha, and cytomix was 1.8, 2.33, and 2.31-fold. Nasal epithelial cells in the HD group showed elevated steady-state iNOS expression even in untreated. Cytokine treatment did not affect the degree of iNOS expression in this group. CONCLUSION: These results confirm our previous findings that nasal epithelial cells in patients with allergic rhinitis produce higher levels of NO through the concomitant expression of different NOS isoforms. We also demonstrated that nasal epithelial cells have the potential to express iNOS protein spontaneously or upon stimulation with inflammatory cytokines, such as IFN-gamma and TNF-alpha. Because the high level of exhaled NO is considered a potential marker of allergic airway inflammation, preserving the iNOS gene from its unregulated induction may be important for maintenance of nasal homeostasis and may offer a tool for therapeutic intervention. PMID- 11905054 TI - [Stem cell factor production from cultured nasal epithelial cells--effect on SCF production by drugs]. AB - We studied whether epithelial cells cultured in serum-free medium contained other cells or not, there were differences in SCF production from cultured nasal epithelial cells between groups of nonallergic and allergic patients, and among degrees of serum mite-CAP RAST classes of allergic patients, and how drugs inhibited SCF production. As a result, no other contaminating cells except mast cell existed in cultured cells. There was a significant difference in SCF production of cultured cells between nonallergic and class 1-2, 3-4, 5-6, and between class 1-2 and 3-4, 5-6 of mite CAP-RAST class. Cyclosporin, prednisolone, fluticasone, ketotifen, and clemastine inhibited SCF production from cultured epithelial cells, but cromoglicate and suplatast did not. Inhibition means the reduction of SCF from cells, not the growth of cultured nasal epithelial cells. PMID- 11905055 TI - [A case of enlarged vestibular aqueduct syndrome with PDS gene mutations]. AB - Enlarged vestibular aqueduct (EVA) is an inner ear anomaly occasionally associated with sensorineural hearing loss (SNHL) and/or dizziness. Recent genetic studies indicate that mutations in the PDS gene may cause EVA. A 10-year old EVA patient who had undergone annual hearing tests for 7 years had an aunt and cousin who also had hearing loss and EVA, so genetic examinations were conducted for a possible genetic link. Two new PDS gene mutations, S610X and S657N, were found in all 3, including the proband. We discuss the importance of genetic analysis, which offers new insight into SNHL diagnosis and treatment in children. PMID- 11905056 TI - [Strategy of neck dissection in parotid cancer]. AB - We conducted definitive surgery on 45 patients with untreated primary parotid cancer from 1975 to 1995, and evaluated methods of neck dissection and results of treatment. All 14 with clinical neck lymph node metastasis underwent ipsilateral radical neck dissection and only 1 developed neck lymph node recurrence at the peripheral dissected site. Of 31 patients without clinical neck lymph node metastasis, 27 of 19 of 36 with high-grade malignancy and 12 of 24 with T3 or T4 did not undergo prophylactic neck dissection and developed latent neck lymph node metastasis in 2 cases (7.4%). Whereas in most cases we achieved good control of the primary site but neck lymph node recurrences occurred, recurrent sites were observed all around the ipsilateral neck and prognosis were very poor if neck dissection was conducted as secondary treatment. Although histopathological diagnosis was considered feasible for predicting occult neck lymph node metastasis, correct diagnostic with fine needle aspiration cytology revealed only 21.8%. Pathological positive lymph nodes in 15 patients who underwent neck dissection were detected all over (level I to V) the ipsilateral neck and the recurrent positive rate at level II was 100%. Based on the above results, we conclude that (1) in cases with neck lymph node metastasis in preoperative evaluation, ipsilateral radical neck dissection is mandated, and (2) in cases without neck lymph node metastasis, prophylactic neck dissection is not usually needed. When pathological results of frozen section from intraoperative jugulodigastric nodal sampling are positive, ipsilateral radical neck dissection is mandated. PMID- 11905057 TI - An evaluation of coronary artery bypass grafting without aortic cross-clamping due to severely atherosclerotic ascending aorta. AB - OBJECTIVES: We evaluated coronary artery bypass grafting (CABG) in which aortic cross-clamping is not done due to severe atherosclerosis of the ascending aorta. METHODS: Subjects were 51 patients undergoing CABG without aortic cross-clamping during cardiopulmonary bypass under moderately hypothermic ventricular fibrillation in the 12 years from June 1988 to October 1999 (Group N). In some cases, empty beating or moderate hypothermic circulatory arrest was used. We compared these 51 with 1104 subjects undergoing conventional CABG with aortic cross-clamping and cardioplegic cardiac arrest in the 9 years from June 1988 to December 1997 (Group A). RESULTS: In all 6 cases with neurologic deficits, moderately hypothermic circulatory arrest was used during proximal anastomosis of saphenous vein grafts. Postoperative computed tomography scan showed them to have suffered infarction due to embolization. Multivariate analysis identified proximal saphenous vein grafting under moderately hypothermic circulatory arrest as a predictor of neurologic deficit. Complete revascularization was significantly lower in Group N. Actual survival and freedom from cardiac death were significantly lower in Group N. CONCLUSION: Manipulation of the atherosclerotic ascending aorta under moderately hypothermic circulatory arrest or ventricular fibrillation generates the highest risk of perioperative neurologic deficit and should thus be avoided. In-situ arterial grafting should be conducted with utmost care. PMID- 11905058 TI - Cosmetic benefits of lower midline skin incision for pediatric open heart operation. A review of 100 cases. AB - OBJECTIVE: Since we developed the procedure in 1996, we have now performed 100 pediatric open heart operations using a lower midline skin incision and a minimal sternotomy approach. METHODS AND RESULTS: To elucidate the benefits of this approach, we analyzed these 100 cases retrospectively. There was no death, and no major complication, caused by this approach, and the resulting scarring in each patient is difficult to be seen under a common undershirt. CONCLUSION: This review shows that the technique of a lower midline skin incision and minimal sternotomy approach is a safe reliable and cosmetically advantageous method for a pediatric cardiac operation. PMID- 11905059 TI - Geometrical difference between an ascending aneurysm and a root aneurysm in valve sparing operations. AB - OBJECTIVE: The aortic root geometry of the leaflet size and coaptation in an ascending aortic aneurysm, and in a root aneurysm, may predict the early and late outcomes from valve-sparing surgery. METHODS: The aortic root was investigated using intraoperative endoscopy before and after valve-sparing root reconstruction. The definition of 'root aneurysm' was marked sinus dilatation proximal to the sinotubular junction. 'Ascending aneurysm' was defined as dominant dilatation distal from the sinotubular junction. Fifteen cases were examined and classified into two groups; Group A with an ascending aneurysm (four patients), and Group R with a root aneurysm (eleven patients). RESULTS: Cusp prolapse was seen in 10 (90.9%) patients of Group R, and in only one (25%) patient of Group A. The length of the free margin of all cusps in Group R was significantly longer than those in Group A. In Group R, the lengths of the free margin of an individual cusp were significantly different, indicating asymmetric cusps. In Group A, the length of the free margin were similar, indicating symmetric cusps. There was no significant difference in the degree of immediate postoperative aortic insufficiency, between the two groups. At the most recent follow-up, progressive aortic insufficiency was present in two patients of Group R. CONCLUSIONS: A root aneurysm had asymmetric, elongated and prolapsed aortic cusps, while an ascending aneurysm had symmetric cusps without prolapse. The mechanism of aortic insufficiency in a root aneurysm was more complex, suggesting a difficulty in achieving long-term valve competence. Such geometrical difference should be considered in the indication for the surgical technique of valve sparing operations. PMID- 11905060 TI - The superiority of pulmonary function after minimally invasive direct coronary artery bypass. AB - OBJECTIVE: We determined whether minimally invasive direct coronary artery bypass (MIDCAB) leads to excellent postoperative pulmonary function, and which contributes more to this--minithoracotomy or avoidance of cardiopulmonary bypass. METHODS: Pulmonary function 1 week before and 2 weeks after surgery was evaluated in 8 patients undergoing MIDCAB (Group M), 10 undergoing off-pump coronary artery bypass (Group O), and 12 undergoing conventional coronary artery bypass grafting (Group C). Parameters were adjusted by their predicted values and postoperative values were expressed as a ratio to preoperative ones. RESULTS: Only Group M maintained postoperative vital capacity and forced expiratory volume in 1 second close to the preoperative level and thus, showed significantly better recovery than Groups O and C. No significant difference was seen between Groups O and C. CONCLUSIONS: MIDCAB provides better recovery of pulmonary function early postoperatively than other procedures thanks to minithoracotomy rather than avoidance of cardiopulmonary bypass. PMID- 11905061 TI - Successfully repaired traumatic tracheal disruption and cardiac rupture with cardiopulmonary support. AB - A 19-year-old man suffering from dyspnea associated with tracheal and cardiac rupture from a traffic accident was found by bronchoscopy to have a 7.5 cm longitudinal tear in the membranous portion of the trachea. Right posterolateral thoracotomy was conducted and open ventilation through the left main bronchus initiated with standby cardiopulmonary bypass cannulation of the right femoral artery and vein. When oxygenation was poor, extracorporeal circulation was initiated through the cannulated artery and vein. Under the cardiopulmonary bypass, we safely repaired the tracheal laceration and cardiac rupture. PMID- 11905062 TI - Chest wall myxoma protruding into the thoracic cavity. AB - Myxoma protruding from the chest wall into the thoracic cavity is very rare. We report our experience in treating a 42-year-old man admitted for a painless mass on the anterior chest wall. Magnetic resonance imaging showed a sharply defined 5 x 4 x 4 cm mass protruding from the chest wall into the thoracic cavity, which we excised surgically. Histologically, the tumor proved to be a myxoma. PMID- 11905063 TI - Bronchial leiomyoma with atelectasis in the left lower lobe. AB - Bronchial leiomyoma with atelectasis in the left lower lobe occurred in a 44-year old woman. The leiomyoma was diagnosed by bronchofiberscopy, and left lower sleeve lobectomy was performed. The histopathology of the tumor revealed a leiomyoma with no evidence of malignancy. An early accurate diagnosis is thought to be important for a more conservative treatment. PMID- 11905064 TI - Beating coronary bypass with right-heart bypass. A safer option in patients at high risk for stroke. AB - Two patients with bilateral obstructive carotid artery disease underwent beating heart coronary bypass including revascularization of the circumflex branch using right-heart bypass in a stable hemodynamic state. Without this mechanical support, lifting the left ventricle for the exposure of the posterior wall could impair the hemodynamic state of the patient. Right-heart bypass in addition to aortic no-touch technique can be a safer option for complete coronary revascularization in patients at high risk for neurological complications. PMID- 11905065 TI - Mediastinal parathyroid cyst with tracheal constriction. AB - A 63-year-old man visiting a physician for slight dyspnea, attributed to a lump on his neck, was found in ultrasonography and computed tomography to have a cyst extending from the left lobe of the thyroid gland to the superior mediastinum. Radiography showed right deviation of the trachea. The cyst disappeared after fine-needle aspiration, but cyst fluid subsequently reaccumulated and he was admitted to our hospital. No abnormalities were detected in tests of thyroid and parathyroid function or blood chemical analysis. The cyst was surgically removed and diagnosed as a nonfunctioning parathyroid cyst, based on the high-intact parathyroid hormone in cyst fluid. The patient recovered fully and has shown no recurrence in the 11 months to data since surgery. PMID- 11905067 TI - 2020 vision: the wish list? PMID- 11905066 TI - Simultaneous repair of stenosis in coronary and vertebral arteries and aortic regurgitation secondary to Takayasu's aortitis. AB - We report a case of severe stenosis in the ostium of both the coronary artery and the proximal left vertebral artery and severe aortic regurgitation secondary to Takayasu's aortitis. A 47-year-old woman underwent simultaneous repair consisting of aortic valve replacement, triple coronary artery bypass grafting, and aorto left vertebral artery bypass. Saphenous vein grafts to 3 coronary arteries and the left vertebral artery were proximally anastomosed on a bovine pericardial patch in the ascending aorta. Since the patient had severe preoperative ischemic symptoms from vertebral-basilar insufficiency, we clamped the vertebral artery during reconstruction under deep hypothermic circulation. The postoperative course was uncomplicated. Simultaneous repair of such multiple lesions requires meticulous planning of surgical procedures and circulatory assist systems. PMID- 11905068 TI - General anaesthesia and sedation post-November 1998. PMID- 11905069 TI - Sedation diploma with SOCS. PMID- 11905071 TI - Letter from the Secretary. PMID- 11905070 TI - What would happen if it were decided that patients on the NHS could only have occlusal amalgams? PMID- 11905072 TI - Advanced life support update. PMID- 11905073 TI - Hot spots in the 2002 OIG work plan. PMID- 11905074 TI - Target: patient safety. PMID- 11905075 TI - How HIM can ease the pain of medical errors. PMID- 11905076 TI - Quality at the core. Interview by Anne Zender. AB - Is your hospital getting ready for the Joint Commission's core measures project? Some hospitals are already transmitting the necessary data as part of the pilot project. Here's how they are handling the challenges and deriving benefits. PMID- 11905077 TI - ICD-9-CM committee proposes new codes, changes. AB - This is part one in a two-part summary of proposals from the November ICD-9-CM Coordination and Maintenance Committee meeting. Part 2 will be published in the April 2002 Journal of AHIMA. PMID- 11905078 TI - HIM's role in monitoring patient safety. PMID- 11905079 TI - [Human recombinant activated protein C in patients with severe sepsis and septic shock]. PMID- 11905080 TI - [Detection of anomalous origin of left coronary artery by spiral CT as a cause of angina pectoris]. AB - The anomalous origin of the left coronary artery can lead to angina pectoris, acute myocardial infarction or even sudden death, especially during exercise. We present a patient in whom the anomalous origin of the left coronary artery is from the right sinus of Valsalva, crossing between the aorta and the pulmonary trunk and causing ischemic chest pain. The anomaly was verified by a Spiral CT, as the coronary angiographic findings were not conclusive, particularly regarding the left course in relation to the major arteries. We suggest that Spiral CT is useful for detecting this kind of anomaly, particularly in clarifying the relationship between the left main coronary artery and the major arteries. PMID- 11905081 TI - [Preliminary results of adult-to-adult living donor liver transplantation: improved size matching and better outcomes with right lobe versus left lobe grafts]. AB - Grafts from living donors are a novel alternative to cadaveric organs for adult liver recipients. METHODS: Twenty adults (mean age, 46 +/- 15 years; mean weight, 68 +/- 12 kg) received left lobes (n = 10; mean weight, 481 +/- 73 g) or right lobes (n = 10; mean weight, 805 +/- 115 g) from living donors (mean age, 38 +/- 10 years; mean weight, 85 +/- 9 kg). Hepatectomy was done using ultrasonic scissors, without vascular isolation. Transplantation was performed in a piggy back fashion. Biliary reconstruction was performed with a single hepatico enterostomy to the left duct for LL grafts and with 2 (n = 8) or 3 (n = 2) hepatico-enterostomies in RL grafts. RESULTS: All donors recovered without significant complications or heterologous transfusion. Three recipients (15%) died. Graft survival is 70% (mean follow-up, 8 months). Mean graft: recipient weight ratios (GRWR) were 0.75% +/- 0.21% (range, 0.51-1.06) in LL recipients, and 1.10% +/- 0.15% (range, 0.93-1.44) in RL recipients. Four recipients of relatively small LL grafts (GRWR < 0.8%), all with Child's B or C cirrhosis, developed small-for-size syndrome. One died; 2 required retransplantation. Small LL grafts were used successfully in Child's A patients or non-cirrhotics (the indication for transplantation was tumor). The LL group had 5 bile leaks (50%) and 3 major vascular complications (30%). One was fatal; 1 mandated retransplantation. The RL group had 2 biliary leaks (20%) and 1 hepatic artery thrombosis, which necessitated retransplantation. CONCLUSIONS: Right lobe grafts prevented small-for-size syndrome. Although donor hepatectomy and recipient grafting are technically more demanding, right lobe grafting seems to involve fewer recipient complications and better outcome. PMID- 11905082 TI - [Video-assisted thoracoscopic lobectomy--preliminary experience]. AB - BACKGROUND: Video-assisted thoracoscopic surgery has become routine worldwide. On the basis of accumulating experience and technologic improvements, more complex operations can be performed. Until recently, thoracoscopic lobectomy has not been performed in Israel. GOALS: To describe the initial experience with thoracoscopic lobectomy in Sheba Medical Center, and discuss the advantages, disadvantages and indications for its use. MATERIAL & METHODS: From June 2000 to January 2001, five patients (3 male, 2 female) 22-72 year-old underwent thoracoscopic lobectomy. Four had malignant neoplasms (3 lung primary) and one a benign process. Preparations for surgery, anesthesia and monitoring were standard as for open lobectomy. Surgery was done through 3 ports and a 5-7 cm auxiliary thoracotomy. The majority of the operation was performed with staplers. In patients with lung tumors the regional lymph nodes were sampled. Pulmonary function tests were evaluated prior to surgery and in the immediate postoperative period. RESULTS: Lobectomy was completed in all patients according to preoperative planning without conversion to open thoracotomy. Operative time was 120-160 minutes. Blood transfusion was not required in any patient during or after the lobectomy. There was no operative mortality and only two minor complications in one patient. Narcotic requirement did not exceed 40 mg of morphine in any patient. Pulmonary function tests revealed a smaller than expected decrease compared to open thoracotomy. The cosmetic results were excellent. CONCLUSIONS: Thoracoscopic lobectomy is feasible in Israel too. It is an acceptable alternative for patients needing this operation. Thoracoscopic lobectomy causes less surgical trauma, better functional result and expectance for lowered mortality and morbidity. Its main disadvantage is its high cost. PMID- 11905083 TI - [Bezoars: from ancient talismans to modern disease]. AB - Bezoars are concretions of foreign materials that are created in the intestinal tract of various animals and humans. They have been known to western medicine since ancient times, when they were used for countering and preventing poisoning. In modern times bezoars were described as an unusual cause for various gastrointestinal symptoms. Trichobezoars are an infrequent form of bezoar, formed from ingested hair. We present a case of a trichobezoar, which appeared as a large abdominal mass, with gastric bleeding, ulceration and perforation. As in most previously reported cases, the diagnosis was not made on clinical grounds prior to imaging studies. PMID- 11905084 TI - [Difficulties in diagnosis and localization of recurrent medullary thyroid carcinoma]. AB - Recurrent and residual medullary thyroid carcinoma (MTC) are common in patients following primary surgical resection. Difficulty arises in performing precise localization of the tumor because of anatomical distortion of the neck structures following surgery. To date, no modality has been shown superior to others in the diagnosis of recurrent or residual MTC, and the issue is currently under debate in the literature. We report a case in which secondary recurrence of MTC was detected and localized using a novel combination of preoperative and intraoperative radionuclide imaging, and a method of preventing intraoperative damage to the recurrent laryngeal nerve in the anatomically disrupted neck. To our knowledge, this is the first report of a combination of these three modalities in detection and localization of recurrent MTC, while minimizing the possibility for nerve injury during the operative procedure. Such a therapeutic strategy may prove useful in the management of patients who have previously undergone neck surgery and suffer from anatomical distortion of normal neck structures. PMID- 11905085 TI - [Plasma anti-oxidants and rheumatoid arthritis]. AB - We present a clinical study aimed to compare plasma antioxidant vitamins, vitamin E, beta-carotene and vitamin A. The study consisted of a group (15 patients) with rheumatoid arthritis (RA) compared to a healthy control group. There was a significant decrease in plasma vitamin E, beta-carotene and vitamin A (vitamin E 30.4 +/- 4.9 VS 43.6 +/- 8.2 micrograms/ml, beta-carotene 0.73 +/- 0.26 VS 1.02 +/- 0.22 micrograms/ml and vitamin A 0.22 +/- 0.07 VS 0.46 +/- 0.15 microgram/ml, P < 0.01 patients VS control, respectively). Supplementation of Dunaliella (natural)--beta-carotene to the RA patients for 3 weeks, resulted in a significant increase in plasma vitamin E (47.9 +/- 5.5 micrograms/ml) beta carotene (0.87 +/- 0.21 microgram/ml) and vitamin A (0.55 +/- 0.15 microgram/ml). There were no changes in the activity indexes of RA. Low plasma antioxidant vitamins in patients with RA are consistent with the observation that oxidative processes occur in the inflammed joints. The validity of antioxidant vitamins as supplementary therapy for RA is not clear. PMID- 11905086 TI - [Lupus-like disease due to minocycline]. AB - Minocycline in another drug that can induce drug-induced lupus. Minocycline is frequently used for acne as a prolonged treatment, so it is important to be aware of the risks in this treatment. The risk ratio for the development of lupus due to minocycline is not known, but it seems to be low. We described a 26 year old female who was treated with minocycline due to acne. The treatment was complicated by rash and serological signs of lupus (antinuclear antibodies and anti-DNA antibodies). The aim of this article is to raise the level of awareness of this complication due to minocycline. PMID- 11905087 TI - [Ricin--from a Bulgarian umbrella to an optional treatment of cancer]. AB - Ricin toxin, found in the bean of the castor plant, is one of the most toxic and easily produced plant toxin. It is composed of two polypeptide chains linked by a disulfide bond. The toxin irreversibly blocks protein synthesis. Oral intoxication is the most frequent mode of exposure, but direct contact and inhalation have also been described. A single intramuscular exposure was reported, related to an assassination event in 1978. The clinical signs of intoxication begin hours after the exposure, and include gastrointestinal signs, dyspnea, fever, vasculatory collapse, neurological signs, liver necrosis and renal failure. Treatment is mainly supportive. A vaccine has been developed and found effective in animal studies. Ricin may be used as a biological weapon due to its heat stability and worldwide availability. A more friendly use, which has been tested in clinical trials, is the incorporation of ricin-immunotoxin for the treatment of cancer patients. PMID- 11905088 TI - [Signal transduction inhibitor--STI571--a new treatment for chronic myeloid leukemia (CML), which opens a new targeted approach to cancer therapy]. AB - Chronic myeloid leukemia (CML), in most of the cases, is the molecular consequence of the t(9,22) translocation, resulting in the Philadelphia (Ph) chromosome and the creation of the fusion gene BCR-ABL. The fusion gene is translated to the protooncogen BCR-ABL, a constitutively activated tyrosine kinase that is linked to the malignant transformation. Thus, this tyrosine kinase became an attractive target for drug design. The development of the novel investigational drug STI 571 is based on its potent and selective ability to inhibit this fusion tyrosine kinase. In preclinical studies, STI 571 selectively inhibited the growth of CML cells that carry the Ph chromosome. In this review we discuss the drug development and design, its mechanism of action, the preclinical studies and the results of phase I and II clinical trials. PMID- 11905089 TI - [Chronic immune-mediated neuropathies]. PMID- 11905090 TI - [Osteoporosis in men]. AB - While many still consider osteoporosis a gender-specific condition of women, an increasing volume of data is accumulating on the epidemiology, pathophysiology and treatment of osteoporosis in men. The main conclusions arising from the available data indicate that the number of osteoporotic men is increasing, and is expected to continue rising due to the increase in life expectancy for men. Risk factors for osteoporotic fractures are similar in men and women. Despite some gender differences in the pathophysiology of osteoporosis, responses to available osteoporosis treatments seem to be the same. Based on available data, it is recommended to implement preventive measures (calcium and vitamin D supplementation) on aging men, similar to women, and to review the official policy on the management of osteoporosis to also include men at high risk. PMID- 11905091 TI - [The prognostic value of the site and extent of Y chromosome microdeletions on spermatogenesis]. AB - Genes on Y-chromosome are involved in the regulation of spermatogenesis. Y chromosome microdeletions were identified among infertile patients in a frequency of 7.5-15% on the long arm of the chromosome. These microdeletions were clustered in 3 main regions named AZFa, AZFb, and AZFc. Reanalyzing the histological findings in men with well-defined varying extent of Y-chromosome microdeletions improved our understanding of the prospect of finding testicular spermatozoa. The chances of finding spermatozoa were almost nil in men with microdeletions that include the complete AZFa region or AZFb region or at least two AZF regions. Large microdeletions that include the Yq tip were suggested to cause chromosomal instability and were shown to be prone to Y chromosome loss. In addition, a decrease of sperm count over time in men with AZFc deletions has been reported and the option of spermatozoa cryo-preservation need to be taken in consideration. Analysis of the Y-chromosome microdeletion was found to be of prognostic value in cases of infertility both in terms of clinical management as well as for understanding the etiology of the spermatogenesis impairment. PMID- 11905092 TI - [Advance medical directives]. AB - A patient's rights to autonomy and to participate in the decision making process is a fundamental ethical principle. However, for the non-competent patient, participation in decision-making is more problematic. A survey carried out in Israel found that less than half of the offspring of terminally ill elderly patients knew the request of their parents regarding life-supporting measures. A solution to this problem is the use of medical advance directives (MADs). In the U.S.A (in 1991) it was required by a federal law to inform every hospitalized patient of his right to use MADs. The experience from the use of MADs in the USA during the last 10 years show that: 1) Most lay persons as well as medical staff support the use of MADs 2) The rate of the use of MADs is about 20%, and among long term care hospitalized patients it is even higher. 3) Sex, age, level of education, morbidity and income were found to be significant factors. 4) Education on the use of the MADs raised the rate of use. 5) Most of the patients who had MADs did not discuss the issue of life supporting treatment with their physicians. 6) Patients who had MADs received less aggressive treatment with reduced medical cost. 7) There is a preference to write generic MADs. Arguments supporting the use of MADs state that they: extend patient autonomy; relieve patient anxiety regarding unwanted treatment; relieve physicians' anxiety concerning legal liability; reduce interfamily conflicts, and they also lower health care costs. Arguments opposing the use claim that they: violate sanctity of life; promote an adversarial physician-patient relationship; may lead to euthanasia; fail to express the patient's current wishes and may even counteract physicians' values. On the basis of experience in the USA and the positive attitude regarding MADs, it appears that MADs can also be applicable in Israel. PMID- 11905093 TI - [Obstructive sleep apnea in children--pathophysiology update]. AB - The obstructive sleep apnea syndrome (OSAS) is a common and potentially serious problem during childhood. Airway obstruction during sleep may occur due to narrowed bony passages in conditions such as congenital craniofacial anomalies or in the collapsible segments of the upper airways (naso/hypo pharynx). Adenotonsillar hypertrophy has been considered as the main and most important causative factor for OSAS in the pediatric population. However, adenotonsillar hypertrophy is not the sole cause for this syndrome. Normal infants and children have relatively narrowed upper airways. They maintain airway patency during sleep by increasing the upper airway neuromotor tone and adjusting central ventilatory drive. Recent studies suggest that childhood OSAS is a dynamic process resulting from a combination of structural and neuromotor abnormalities, rather than from structural abnormalities alone. Genetic factors have also been suggested. This review delineates the current concepts of the pathophysiology of OSAS in infants and children. PMID- 11905094 TI - [Update on oral cancer--awareness equals survival?]. AB - There is a trend towards higher prevalence of oral cancer and no progress has been achieved concerning survival rates during the last few decades. Despite the simplicity of the oral examination most of oral cancers are discovered at advanced stages bearing severe prognosis. Improved awareness of both the attendant medical team and the target population may improve the chances of prevention by earlier detection thus enhancing the survival rate. We present current diagnostic procedures for early detection of oral cancerous lesions together with a short review of epidemiology, clinical aspects, risk factors and treatment modalities. PMID- 11905095 TI - [Chylomicronemia syndrome]. AB - The chylomicronemia syndrome is a disorder characterized by severe hypertriglyceridemia and massive accumulation of chylomicrons in plasma. This hypertriglyceridemia can lead to the development of eruptive xanthomas, lipemia retinalis, and is clinically important when plasma triglyceride levels predispose to pancreatitis (above 2000 mg/dl). Three genetic disorders have been described in which chylomicrons accumulate in plasma: familial lipoprotein lipase deficiency, familial apolipoprotein C-II deficiency, and familial inhibitor to lipoprotein lipase. In addition, chylomicronemia is seen in other states with the simultaneous occurrence of familial forms of moderate hypertriglyceridemia and other acquired causes for hypertriglyceridemia such as diabetes mellitus, certain drug therapies and alcohol use. Treatment should be directed at both the familial and the acquired disorder. This review discusses the chylomicronemia syndrome presenting the pathophysiologic characteristics of triglyceride and chylomicrons metabolism, diagnosis, prevalence and treatment. PMID- 11905096 TI - [Multi-organ failure in a young soldier: a clinical-pathological meeting]. AB - This clinical-pathological conference took place at the Sourasky Medical Center, Tel Aviv, on February 21, 2001. We present the case of a young and previously healthy soldier who developed multi-organ failure with predominant liver dysfunction following exertional heatstroke. The patient's clinical course consisted of an early phase of transient encephalopathy, associated with hyperthermia, hypophosphatemia, mild laboratory indications of renal failure, rhabdomyolysis and consumption coagulopathy. Following an intermediate convalescing phase that lasted a single day the patient deteriorated into a catastrophic course with hemodynamic instability, fulminant hepatic failure, respiratory distress, kidney failure, rhabdomyolysis, coagulopathy and coma. He died 4 days later. In this article we elaborate on the association of heatstroke with multiple organ dysfunction syndrome in general, and fulminant liver failure in particular. The nature of hypophosphatemia and the possible role of additional injury from acetaminophen are discussed. PMID- 11905097 TI - [The amputated leg--a tale of scientific curiosity--1792]. AB - In 1792 the young military surgeon Dominique Jean Larrey (later Baron de l'empire) had to amputate a soldier's leg. His scholarly knowledge combined with intellectual curiosity turned a common event into an innovative scientific experiment, after he used the severed leg to affirm in the human being the recent observations made by Galvani on frogs. The possibility of inducing muscular contractions by galvanic current led him to foretell, much ahead of his time, that this mode would facilitate rehabilitation of the paralyzed. PMID- 11905098 TI - [Feeding a terminal patient, sustaining treatment and respecting the right to die]. AB - Great changes have occurred in the doctor-patient relationship--in medical and social scopes. Lately, there are deliberations regarding whether to withdraw nutrition from a terminal patient. This question raises the basic issues of respecting the patient's wish and his autonomy. Nourishment through the digestive system may prolong life; however, the decision concerning this dilemma differs from country to country. The Jewish religion has its own position on the matter. The patient knows the doctor has no conflict between trying to help or kill and continues to fully trust the physician. Whenever nourishing or treating a terminal patient, the doctor should be assisted by the family's decision or a psychiatric resolution. Actually, there is rarely conflict between the evaluation of the treating physician, the position of the family and the court's decision. PMID- 11905099 TI - General hospitals. Acute anxieties. AB - Acute hospitals have suffered at least 15 years of neglect. Little attention has been given to the key role of support services. The structure of trusts hinders the development of clinical networks. The way consultants are employed hampers their involvement in the modernization agenda. PMID- 11905100 TI - Consultants' hours. A day in the life. AB - Many NHS consultants are working more than their contracted hours. Diaries kept by 32 consultants in one trust revealed an average working week of 50 hours. A pilot scheme rescheduling the work patterns in a haematology department to include a rest day has reduced the number of hours worked. The scheme is popular with consultants and could be transferable to other teams. PMID- 11905102 TI - Has the space age stalled? PMID- 11905101 TI - Distinction awards. Prize guys. AB - The distinction awards pay scheme for consultants has been the subject of controversy since its introduction in 1948. It has lacked transparency and has been criticised for being inequitable. Many academic consultants receive awards without fulfilling the required service commitment. Chief executives are turning a blind eye to this. Proposals for a revised scheme, due to take effect in April 2003, only address some of the shortcomings. PMID- 11905103 TI - Fertility. Looking at ART. PMID- 11905104 TI - Aviation. Heads on tails. PMID- 11905106 TI - It's not easy being green. Developing environmentally safe products is one thing; marketing them is another matter entirely. PMID- 11905105 TI - Immunology. Drink your shots. PMID- 11905108 TI - Augmented reality: a new way of seeing. PMID- 11905107 TI - Proteins rule. PMID- 11905109 TI - Parasitic sex puppeteers. PMID- 11905110 TI - Ripples in spacetime. PMID- 11905112 TI - The social psychology of modern slavery. PMID- 11905111 TI - The science of bad breath. PMID- 11905113 TI - Grow, then kill. PMID- 11905115 TI - Weathering a political storm. A contextual perspective on a psychological research controversy. AB - In the spring of 1999, a storm of controversy arose at the local, state, and national levels surrounding an article on the effects of child sexual abuse published in 1998 in Psychological Bulletin. The article was vehemently denounced by various media outlets, conservative grassroots organizations, members of the general public, state legislatures, and ultimately by the United States Congress. The authors chronicle these unprecedented events and related challenges faced by the American Psychological Association. The authors also describe the Association's efforts to resolve the crisis, while staunchly upholding academic freedom and scientific integrity, and review the lessons learned for the field of psychology. PMID- 11905114 TI - Exploring a controversy. PMID- 11905116 TI - When worlds collide. Social science, politics, and the Rind et al. (1998). Child sexual abuse meta-analysis. AB - A 1998 meta-analysis by B. Rind, P. Tromovitch, and R. Bauserman in Psychological Bulletin indicated that the relations between child sexual abuse and later psychopathology were weak in magnitude. Shortly thereafter, this article was condemned by media personality Dr. Laura Schlessinger and numerous conservative organizations and was denounced by the United States Congress. In addition, the American Psychological Association (APA) distanced itself from the authors' conclusions. This incident raises questions regarding (a) authors' responsibilities concerning the reporting of politically controversial findings, (b) academic and scientific freedom, (c) the role of the APA in disabusing the public and media of logical errors and fallacies, and (d) the substantial gap between popular and academic psychology and the responsibility of the APA to narrow that gap. PMID- 11905117 TI - Politics, operant conditioning, Galileo, and the American Psychological Association's response to Rind et al. (1998). AB - The controversy surrounding B. Rind, P. Tromovitch, and R. Bauserman (1998) provides valuable lessons into scientific independence, politics, and organizational decision making. In an unprecedented action, the U.S. Congress officially condemned findings of Rind et al. Meanwhile, the American Psychological Association took similarly unprecedented measures in an effort to assuage its Congressional critics. This article, written from the perspective of a psychologist serving in Congress, discusses the various political and organizational dynamics that developed during the controversy. Understanding and learning from this incident can help psychologists and their professional associations better prepare for and respond to potential controversies arising from research or other publications. PMID- 11905118 TI - Everything you need to know to understand the current controversies you learned from psychological research. A comment on the Rind and Lilienfeld controversies. AB - Psychological theory and research can help explain some of the aspects of the controversies that arose over articles written by B. Rind, P. Tromovitch, and R. Bauserman (1998) and by S. O. Lilienfeld (2002). In particular, one needs to distinguish between rational and intuitive thinking, to recognize the power of context, to be reflective in one's own thinking, and to realize the costs of defying the crowd. There are steps one can take to be wiser and more balanced in one's own thinking than one may have been in the past. One such step is to resolve conflicts among psychologists by attempting to defuse rather than to exacerbate or avoid them. PMID- 11905119 TI - Science, politics, and peer review. An editor's dilemma. AB - The American Psychologist is the official journal of the American Psychological Association. As such, it is a valued outlet for articles dealing with reviews of current topics in psychology, policy issues, and critiques of current research. S. O. Lilienfeld submitted a manuscript to the journal that was accepted by the ad hoc action editor; however, the action editor's decision was later overruled by the editor, and additional changes to the manuscript were requested. Because of this editorial decision, a controversy arose that played out on various Internet discussion groups. The author presents his perspective as the editor in this controversy. Points of emphasis include the need to protect the integrity and the confidentiality of the peer review process for scientific journals. PMID- 11905121 TI - Publication of Rind et al. (1998). The editors' perspective. AB - The authors address several issues surrounding the B. Rind, P. Tromovitch, and R. Bauserman (1998) meta-analysis on the long-term effects of childhood sexual abuse from their perspective as the editors who accepted the manuscript for publication. In particular, they discuss the appropriateness of their editorial decision, the appropriateness of a policy of considering authors' prior publications in editorial decisions (as suggested by some critics), and the editors' role in the specific recommendations made by B. Rind et al. They go on to consider actions they could have taken to minimize the mischaracterizations of the study's findings and conclusions and their views of the American Psychological Association's actions with respect to the authors personally and to society more generally. PMID- 11905120 TI - Five commandments for APA. AB - The author delineates 5 rules of scientific review and publishing and argues that these norms need to be upheld even when to do so proves politically difficult. The 5 rules are: (a) Scientific articles should be judged only by their logic and the strength of their evidence; (b) the results of a competent peer review should be accepted; (c) disagreements with scientific articles should be aired in peer reviewed commentaries; (d) efforts to judge scientific articles on the basis of political concerns should be resisted; and (e) the explicit rules and normative expectations of peer review should not be arbitrarily altered. PMID- 11905122 TI - The publishing dilemma of the American Psychological Association. PMID- 11905123 TI - American Psychologist Task Force report. Clarifying mission, coverage, communication, and review process. PMID- 11905124 TI - Challenges and opportunities in the psychological sciences. AB - In this commentary, the author draws on his experiences at the National Science Foundation to reveal that threats to the value and autonomy of social science research are more common than most psychologists suspect. To reduce the likelihood of these threats in the future, it is necessary to improve the public's understanding and confidence in the psychological sciences. The key is to focus on the results and accomplishments of the research and to avoid caveats and qualifications. It is also important to motivate the interest of the public by helping them to understand why and how psychological research is improving the quality of their lives. The author concludes with some practical suggestions regarding how psychologists can become more proactive in their education of the public, the media, and congressional representatives. PMID- 11905125 TI - Collisions, logrolls, and psychological science. AB - Relationships among science, public policy, and the media have long been a topic of controversy. A discussion of this controversy serves to place views on the Rind et al. affair in a broader context and set the stage for constructing more effective working relationships between scientists and both policy and media experts. To advance these relationships, the author offers several recommendations that emphasize ongoing institutional activities and encourage collaboration with other professional organizations. PMID- 11905126 TI - Trial by Internet: cybercascades and the Lilienfeld case. PMID- 11905127 TI - A funny thing happened on the way to my American Psychologist publication. PMID- 11905128 TI - Evaluation of the Utah student injury reporting system. AB - The Utah Student Injury Reporting System (SIRS), implemented in 1984 to monitor injuries to students in grades K-12 in Utah schools, has served as a model for surveillance systems created by other states and some European countries. This paper evaluates the Utah experience in developing and administering the SIRS. The evaluation identifies usefulness of the system, discusses the sensitivity of the system in detecting school injuries, estimates the system's costs, and provides suggestions to other states and districts interested in building a cost-effective and efficient surveillance instrument. PMID- 11905129 TI - Hearing conservation education programs for children: a review. AB - Prevalence of noise-induced hearing loss (NIHL) among children is increasing. Experts have recommended implementation of hearing conservation education programs in schools. Despite these recommendations made over the past three decades, basic hearing conservation information that could prevent countless cases of NIHL remains absent from most school curricula. This paper reviews existing hearing conservation education programs and materials designed for children or that could be adapted for classroom use. This information will be useful as a resource for educators and school administrators and should encourage further development, implementation, and dissemination of hearing conservation curricula. The overall, and admittedly ambitious, goal of this review is to facilitate implementation of hearing conservation curricula into all US schools on a continuing basis. Ultimately, implementation of such programs should reduce the prevalence of noise-induced hearing loss among children and adults. PMID- 11905131 TI - Development of the social ecology model of adolescent interpersonal violence prevention (SEMAIVP). AB - The Social Ecology Model of Adolescent Interpersonal Violence Prevention (SEMAIVP) was used to examine adolescent beliefs and behaviors about interpersonal violence. Cross-sectional data were collected from middle school students using the Youth Conflict Mediation Survey. Regression models were developed for the total sample, by gender and by race. Factors explaining 48% of the variance in violence avoidance responses include more frequent positive recognition for violence avoidance, possessing a higher sense of nonviolence self efficacy, and more frequent involvement in violence engagement behaviors. Factors explaining 53% of the variance in violence engagement responses include more exposure to neighborhood fighting, fewer prosocial anger control beliefs, fewer prosocial anger control strategies, more skill development opportunities, and more frequent involvement in violence avoidance behaviors. Findings suggest that prevention approaches need to use a comprehensive social ecology approach that includes intrapersonal, personal, school, and neighborhood environmental components and that provides recognition to adolescents for using violence avoidance skills. PMID- 11905130 TI - Fifth through eighth grade longitudinal predictors of tobacco use among a racially diverse cohort: CATCH. AB - CATCH provides multiethnic cohort data from third to eighth grades from four US geographic regions. This study examined smoking behaviors and predictors from fifth and eighth grades by ethnicity, gender, and geographic location through self-report data obtained from the cohort (N = 3,654). Overall, eighth grade prevalence for ever smoked was about 44%, 30-day prevalence was about 20%, 7-day prevalence 13.3%, and daily prevalence 7.4%. Prevalence was similar for Caucasians (21.5%) and Latinos (21.6%) and lowest for African Americans (13.1%). The 30-day prevalence for smokeless tobacco was higher for boys than for girls (9.8% vs 5.1%). Tobacco use by parents, siblings, and friends, and easy accessibility in the home in fifth grade, were significant predictors for smoking in eighth grade. Results did not differ by race, gender, or geographic location. The strongest correlate of smoking in eighth grade was having a best friend who smoked. Intention not to smoke in fifth grade predicted nonsmoking in eighth grade. Predictor strength across ethnic groups in different geographic regions was impressive. The social environment of young people continues to be an important instigator of smoking onset. The connection between intention and behavior over time suggests students' intentions not to smoke reflect decision making at an early age. PMID- 11905132 TI - Using path analysis to examine adolescent suicide attempts, life satisfaction, and health risk behavior. AB - This study determined if differences existed between four race/gender groups in regard to attempted suicide among a randomly selected, cross-sectional population of 4,565 public high school students in South Carolina. A modified Youth Risk Behavior Survey was designed to gather information on quality of life, life satisfaction, and six risk-behavior categories. Data first were analyzed using logistic regression analysis and subsequently analyzed using path analysis. Results suggest several independent variables (feelings of intimidation, alcohol and cocaine use, self-perceptions of mental health, self-perceptions of body weight, dieting practices, bulimic episodes, and physical and sexual abuse) were associated significantly (p < .01) with adolescent attempted suicide either directly or indirectly through mediating variables. Significant associations among risk behaviors, mediating variables, and self-reported attempted suicide varied across the four race/gender groups, indicating a need to further study differences noticed in each race/gender scheme. PMID- 11905133 TI - Fast food sales on high school campuses: results from the 2000 California high school fast food survey. PMID- 11905134 TI - Adapting and using intensity measurement in school nursing. PMID- 11905136 TI - Testing your diagnostic skills (#58). Case no. 1. Squamous papilloma. PMID- 11905135 TI - Health on the Net Foundation: advocating for quality health information. PMID- 11905137 TI - Testing your diagnostic skills (#58). Case no. 2. Radicular cyst. PMID- 11905138 TI - More hot topics in HIV/AIDS. PMID- 11905139 TI - The once-a-day era is upon us. PMID- 11905140 TI - NIAID and Merck to collaborate on HIV vaccine development. PMID- 11905141 TI - A clinical challenge in Sagamu, Nigeria. PMID- 11905142 TI - HIV disease management: new technologies improve outcomes and contain costs. AB - HIV disease presents a continuous learning curve as we move from managing a fatal illness to treating a chronic disease. Even though the HIV epidemic is more than 20 years old, our understanding of the disease and its management are constantly evolving. New technologies are an important part of this learning curve. As they are introduced, providers and managed care organizations need to develop policies and procedures that reflect the state of the art in HIV care to continue to build on the good outcomes seen since the advent of HAART. While this model of care is "expensive," it is less expensive long-term than the poorer outcomes and costs associated with increased hospitalization, drug resistance, and preventable morbidity in HIV disease. PMID- 11905143 TI - Progression of HIV-associated dementia treated with HAART. AB - A consecutive series of 96 patients with HIV-associated dementia treated with HAART were studied to identify specific clinical factors associated with an improved response to therapy. The Memorial Sloan-Kettering dementia severity scale and the HIV Dementia Scale were used to assess outcomes. Of 30 patients meeting the inclusion criteria with adequate follow-up, 60% improved neurologically and 40% progressed. There was a trend toward improvement associated with plasma viral suppression, whereas progression was strongly associated with injection drug use history (odds ratio, 13.3). Age, ethnicity, gender, adherence, and predicted CNS penetrance of HAART were not associated with improved outcomes. PMID- 11905144 TI - Primary Nocardia osteomyelitis as a presentation of AIDS. AB - Human infection with Nocardia species presents as a wide range of clinical syndromes. Nocardia is an important opportunistic pathogen in immunocom-promised patients. We report a case of primary Nocardia asteroides osteomyelitis as the initial clinical presentation of AIDS. The infection was successfully treated with a prolonged course of trimethoprim-sulfamethoxazole in conjunction with HAART. Nocardia osteomyelitis should be recognized as an unusual but important and treatable opportunistic infection in patients living with HIV infection. PMID- 11905145 TI - A case of nocardia osteomyelits--is HIV to blame? PMID- 11905146 TI - Postexposure prophylaxis. PMID- 11905147 TI - Effects of central infusions of neuropeptide Y on the somatotropic axis in sheep fed on two levels of protein. AB - Effects of infusions of neuropeptide Y (NPY) into 3rd ventricle of growing sheep fed on diets containing restricted (R) or elevated (E) levels of protein on the immunoreactive (ir) somatostatin neurones, ir somatotrophs, growth hormone (GH) concentration in the blood plasma were studied. The long-term restriction of protein in the diet elicited: enhancing irSS content in periventricular perikarya; diminishing irSS stores in the median eminence and elevating the number ir somatotrophs and content of irGH. NPY infusions enhanced the content of irSS in perikarya in sheep fed on E diet and diminished the number of ir somatotrophs and content of irGH of sheep fed on R diet. The R diet as well as NPY infusions caused an increase in GH mean concentrations in the blood plasma. Obtained results suggest that stimulatory effect of restricted feeding and/or NPY action on GH secretion can be due to attenuated SS output. Since dietary restrictions and exogenous NPY have similar influence on the activation of GH secretion, we suggest that NPY could be a neuromodulatory link between nutritional cues and somatotropic axis in sheep. PMID- 11905149 TI - Changes in short-term potentiation of hypoglossal activity by modulators of nitric oxide production in the rabbit. AB - Stimulation of the superior laryngeal nerve (SLN) causes a potentiation of hypoglossal nerve activity persisting after cessation of stimulation. The mechanism of this phenomenon is uncertain. We investigated a potential role of the nitric oxide (NO) pathway in modulation of the after-effects of SLN stimulation on phrenic and hypoglossal activity in rabbits. L-Arginine, a substrate for NO synthesis and NG-Nitro-L-Arginine (L-NNA) an inhibitor of NO synthase (NOS), were administered systemically. L-Arginine and L-NNA alone caused small changes in respiratory activity. During pre-treatment with NO precursor the amplitude and duration of hypoglossal potentiation evoked by SLN stimulation were reduced. Systemic NO synthase inhibition partially reversed these effects of L Arginine. The results showed that interference with NO production by NO substrate and NOS inhibitor modulates the effects of SLN stimulation on hypoglossal activity. Nitric oxide might be a negative modulator of the transmission of short term potentiation (STP) in hypoglossal activity. PMID- 11905148 TI - Memory effects of arginine vasopressin (AVP) and [7-9] fragment of its peptide chain in rats. AB - The effect of arginine vasopressin (AVP) and [7-9]-vasopressin on different memory types was examined in tests of active avoidance, passive avoidance and water maze after both intraventricular and subcutaneous injections. Results were compared with AVP and [7-9]AVP effects in open field and holeboard tests. Significant latency time elongation was observed after both icv and s.c. injections of AVP but not [7-9]AVP in the passive avoidance test. This effect was blocked by [beta-Mercapto-beta, beta-cyclopentanometyleno1-propionylo1-O-Et Tyr2,Val4,Arg8]-vasopressin, an antagonist of V1 receptors. Injections of both peptides did not improve the performance in other performed tests including the open field test and the holeboard test. Therefore the results confirmed that AVP improves memory only in the passive avoidance test. This effect is probably mediated by central V1 receptors and is not induced by exploratory and locomotor activity impairment. Considering the above results together with the literature data, it seems that the most important fragment of AVP is [4-6] AVP. PMID- 11905151 TI - Event-related current density in primary insomnia. AB - Using Low Resolution Electromagnetic Tomography (LORETA), event-related current density was investigated in 14 patients with primary insomnia and 14 controls matched for age, gender and education level. All subjects were rated on the Athens Insomnia Scale, the Hyperarousal Scale, the Hamilton Depression Rating Scale and the Beck Depression Inventory. They also completed the Selective Reminding Test and the Continuous Attention Test. Only minor elevations on depression scales were found in patients. The Continuous Attention Test did not reveal any between group differences. However, insomniacs required more trials before all the Selective Reminding Test items were learned. Insomniacs showed less event-related current density in orbitofrontal, medial prefrontal and anterior cingulate cortex, i.e. brain regions of relevance for cognition and affect. Earliest group differences appeared in the P1 time range and then were observed at the N1, N2 and P3 stages of stimulus processing. These stimulus processing differences correlated most consistently with severity of insomnia. Neuropsychological impairment correlated most strongly with less current density in Brodmann area 10. PMID- 11905150 TI - Daily GnRH and LH secretion in ewes is not modified by exogenous melatonin during seasonal anestrus. AB - The effect of central, short-term melatonin administration on daily GnRH and LH secretion was studied in ewes during seasonal anestrus. Melatonin, in a total dose of 32 micrograms and the vehicle were perfused for 4 hours into the mediobasal hypothalamus/median eminence (MBH/ME). The mean GnRH concentration during perfusion with melatonin decreased significantly (P < 0.05), as compared to the concentration during the preceding perfusion with the vehicle only. This change resulted from high variations in GnRH concentration noted during the initial phase of perfusion rather than from an action of melatonin. Melatonin perfused into the MBH/ME did not significantly affect LH secretion. A higher dose of melatonin and vehicle were also infused intracerebroventricularly (icv.) in either intact (300 micrograms for 3 hours) or ovariectomized (OVX) ewes (400 micrograms for 4 hours, 100 micrograms/100 microliters/h). In the intact animals, melatonin did not significantly affect LH secretion. Interestingly, melatonin significantly decreased (P < 0.05) the number of LH peaks in OVX ewes. These results demonstrate that melatonin delivered for a few hours directly into the central nervous system did not affect either daily hypothalamic GnRH release or pituitary LH secretion in intact ewes during seasonal anestrus, but did modify pulsatile LH secretion in ewes deprived of the negative feedback of estradiol. PMID- 11905152 TI - Reduced degree of long-range phase synchrony in pathological human brain. AB - In this paper multivariate spontaneous EEG signals from three broad groups of human subjects--control, seizure, and mania--were studied with the aim of investigating the possible effect of these pathologies on the degree of phase synchronization between cortical areas. The degree of phase synchrony was measured by two recently developed measures which are more suitable than classical indices like correlation or coherence when dealing with nonlinear and non-stationary signals like the EEG. Signals were reduced to seven frequency bands (delta, theta, alpha-1, alpha-2, beta-1, beta-2 and gamma) which were statistically compared between the normal and the other two groups. It was found that the degree of long-range synchrony was significantly reduced for both pathological groups as compared with the control group. No clear differences were found in the degrees of short-range synchrony. PMID- 11905153 TI - Electrophysiological investigation of spino-olivary projections originating from sacral segments of the cat spinal cord. AB - Ascending projections of sacral spinal cord neurones (S1-S2) to the dorsal accessory olivary nucleus (DAO) were electrophysiologically investigated in 3 adult cats under deep alpha-chloralose anaesthesia. Antidromic action potentials were recorded extracellularly from 19 cells following stimulation of their axons in both the contralateral dorsal accessory olivary nucleus (coDAO) and the contralateral lateral funiculus at the level of lower thoracic segments (Th13). Two groups of neurones were identified in the gray matter of S1-S2 segments: one distributed in the medial part of Rexed's laminae VI and VII (n = 5), the other located in the ventromedial part of lamina VIII (n = 14). Axonal conduction velocities of neurones investigated were comprised in the range 32-55 m/s. A significant decrease of conduction velocity was observed in each case when distal and proximal parts of the axon were compared. Our research confirmed anatomical data concerning spino-olivary neurones originating from sacral segments. However, we suggest that axons of this pathway give off collaterals to other spinal or supraspinal centres. PMID- 11905154 TI - Pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates phosphoinositide metabolism and translocation of protein kinase C in chick cerebral cortex. PMID- 11905155 TI - The sense of beauty. PMID- 11905156 TI - Clinical assessment of precision of fit. PMID- 11905157 TI - Clinical considerations for enhancing the success of implant-supported restorations in the aesthetic zone with delayed implant placement. AB - Several factors (e.g., root configuration, periapical pathoses, periodontal disease, severe bone loss, patient accessibility) determine the timing of the implant placement. When delayed implant placement is the only feasible option for implant restoration, achieving a functional and aesthetic restoration is more complex and challenging. This article will discuss the surgical and prosthetic methods for enhancing the success of an implant-supported crown in the aesthetic zone in cases where delayed implant placement is the only available treatment option for patients missing single teeth. PMID- 11905158 TI - Prosthodontic restoration of vertical dimension with all-ceramic crowns. PMID- 11905159 TI - Recording and communicating shade with digital photography: concepts and considerations. PMID- 11905161 TI - Is there a "bargain" out there? PMID- 11905160 TI - Anatomical form defines color: function, form, and aesthetics. AB - Contemporary composite materials enable the reproduction of polychromatic effects within a tooth. A broader definition of color that incorporates the anatomy and optical properties of a tooth must be developed so the dental professional can better understand the infinite possibilities of color that exist within the tooth and restoration. This article describes a direct protocol for the development of natural restorations in the posterior dentition through the integration of function, form, and color. PMID- 11905162 TI - Precision shade technology: contemporary strategies in shade selection. AB - Precise color communication is integral to the development of aesthetic harmony and overall restorative success. While traditional shade-taking procedures have enabled a degree of shade transfer, contemporary digital shade analysis devices allow standardized, repeatable shade determinations for increased accuracy. This article presents a clinical case report and contrasts two technology-based shade selection systems (SpectroShade, MHT International, Newtown, PA; ShadeScan, Cynovad, Montreal, Canada) in the retrospective verification of shade during the restoration of a maxillary right central incisor compared to conventional methods. PMID- 11905164 TI - U.N. officials urge effort to reach $10 billion goal annually for AIDS fight. PMID- 11905165 TI - Bush AIDS budget criticized; report cites program waste. PMID- 11905163 TI - Direct posterior composite restorations: simplified success through a systematic approach. AB - Posterior resin-based composites have become an indispensable part of the aesthetic restorative armamentarium. The creation of a functional, anatomical contact, however, remains a challenge for many clinicians. In order to meet both aesthetic and functional demands in the posterior quadrant, a composite resin with enhanced physical and handling properties must be used. This article demonstrates a predictable technique for creating proximal contact using a resin microfill that will allow clinicians to gain confidence in their ability to provide aesthetic and functional Class II restorations. PMID- 11905166 TI - Hepatitis C workers' comp case raises questions for HIV. PMID- 11905167 TI - State budgets. Revenue shortfalls, expenditure overruns result in gaps. PMID- 11905168 TI - Medicaid. Tennessee seeks approval to revise managed-care waiver. PMID- 11905169 TI - Powell criticized for public support of condom use. PMID- 11905170 TI - Report: early sexual activity poses threats to minority youth. PMID- 11905171 TI - Initiative seeks funding, support for effective microbicides. PMID- 11905172 TI - Suit against hospital fails because of lack of actual HIV risk. PMID- 11905173 TI - Man convicted of viatical fraud wins reduction in restitution. PMID- 11905174 TI - Fired nurse did not seek reasonable accommodation for HIV. PMID- 11905175 TI - HIV-positive teen receives $90,000 in bias case settlement. PMID- 11905177 TI - Immigration. Court affirms HIV-positive woman's smuggling conviction. PMID- 11905176 TI - United Airlines employee loses HIV employment bias claim. PMID- 11905178 TI - [Comparative radiographic analysis of surgical and conservative treatment of unstable injuries of the thoracic and lumbar spine]. AB - INTRODUCTION: Treatment of fractures and dislocations of the thoracolumbar spine is aimed at achieving a stable and painless spine. Concerning mode of treatment therapists are divided into two groups--those for conservative approach and those who advocate operative stabilization of spinal column. The objective of this paper was to compare radiological findings of both modes of treatment on the basis of our clinical material. MATERIALS AND METHODS: The examination included radiographs of 96 patients with radiolographic signs of unstable spine, treated at Clinic of Orthopaedic Surgery and Traumatology of the Institute of Surgery of the Clinical Center in Novi Sad from 1984 to 1998. Among 48 operated patients (average age of 40.7), there were more females--25, and among nonoperated, there were more males--33. The average age of conservatively treated patients was 52. Dynamic radiographs evaluated the angle of local kyphosis on the spot of injury (Cobb's angle), reduction of dimensions of the body of injured vertebra and sagittal movement of the body on affected level. Measured values were corrected by values of sagittal index. RESULTS: Initially the average angle of local kyphosis among operated was 26.9, among conservatively treated it was 19.7. Control values for operated were 14.7 and for conservatively treated 25.6. Initially, a wedge-shaped deformity larger than half of the front part of the injured vertebra body had 42 nonoperated and 32 operated patients. Initially, the average sagittal movement of the injured vertebra body among operated patients was 6.97 mm, and at control 1.34 mm. Among nonoperated, the average movement of the body was 1.71 mm initially and 3.23 mm at control. DISCUSSION: Similar results of angular deformity were found by other examiners. Their results are also in favor of operative treatment of unstable injuries of Th-L spine. The number of those proclaiming good results of conservative treatment of injures of thoracolumbar spine in patients with initially high values of angular deformity is much less. Willen, Mumford, Andreycik and others using wedge-shaped deformity and sagittal movement of the body as parameters in their studies had similar results as those in this study, and they conclude that the stability of segment is much higher among operated patients and that operative treatment finally ends in better radiological results. CONCLUSION: Radiological results of operative treatment of unstable injuries of thoracolumbar and lumbar spine are better than those of conservative treatment. In order to obtain a stable spine, which is the aim of treatment of unstable spine injuries, operative stabilization is necessary. PMID- 11905179 TI - [Quality of life assessment in school-age children and adolescents in Yugoslavia from the viewpoint of the children and their parents]. AB - INTRODUCTION: Quality of life assessment among children is relatively new and a poorly investigated concept. Previous results of investigations indicated that parents and children have different perceptions. The main aim of this study was to review health and quality of life components among schoolchildren and youth, from the aspect of pupils and their parents, and to determine differences in assessment of the investigated concept between children and their parents. MATERIAL AND METHODS: The study was performed on a random sample of children and youth (No 600) and their parents. The assessment model was a Yugoslav adaptation of CHQ-CF87 and CHQ-PF50. DISCUSSION AND RESULTS: There is still no adequate answer to the question "who is the most appropriate person to be examined". Over 50% of studies are based on parental assessment. Apart from CHQ-CF87 we have used "parent version" of the questionnaire (CHF-PF50). Comparison of mean values of each scale of both versions of questionnaires significant correlation was established (between 0.31-0.50). However, by investigation of differences between mean values, a significant variation regarding components of health and quality of life between children and their parents was established in 8 out of 10 scales analyzed. CONCLUSION: In our study children valued components of health and quality of life as worse in regard to their parents. Significant differences were established in 8 out of 10 scales analyzed. Significant correlation between children's and parental answers was reached by comparative analysis. The correlation coefficient was 0.31-0.50. PMID- 11905180 TI - [Surgical trauma in laparoscopic and classical cholecystectomy]. AB - INTRODUCTION: Accidental or surgical trauma stimulates a response and its intensity is proportional to extent of trauma. The aim of this prospective study was to compare the intensity of the acute-phase reaction and metabolic changes in patients undergoing elective cholecystectomy for chronic cholecystitis. PATIENTS AND METHODS: Sixty patients with cholelithiasis were divided into two groups: thirty patients underwent laparoscopic cholecystectomy (LC group) and thirty patients open cholecystectomy (OC group). Glucose concentration, mean cortisol concentration, C-reactive protein, albumin levels and lactate-dehydrogenase activity were measured preoperatively and postoperatively for up to 48 hours. RESULTS AND DISCUSSION: The examined groups of patients were comparable in regard to age and sex. The duration of operation was similar in both groups. Postoperative hospital stay after laparoscopic operation was significantly shorter than after open cholecystectomy (p < 0.05). The mean glucose concentration (s.e.m.) during the initial 24 hours after surgery was significantly higher (p < 0.05) following open cholecystectomy. The mean cortisol concentration was significantly higher (p < 0.05) following open in regard to laparoscopic operation. Increase in plasma C-reactive protein was significantly higher (p < 0.05) after open cholecystectomy, with maximal levels 48 h after operation. There was a statistically significant decrease (p < 0.05) in albumin concentration after open cholecystectomy. Serum concentration of intracellular enzyme lactate-dehydrogenase increased significantly (p < 0.05) following open in regard to laparoscopic cholecystectomy. CONCLUSION: According to these results, aspects of metabolic and acute-phase responses and tissue damage are reduced following laparoscopic cholecystectomy. The mean postoperative hospital stay was significantly shorter after laparoscopic cholecystectomy with rapid patient recovery. PMID- 11905181 TI - [Correlation of subjective and objective methods of evaluating hearing threshold]. AB - INTRODUCTION: Objective evaluation of hearing threshold and detection of hearing impairment are the basic prerequisites for hearing and speech rehabilitation. In infants and early pre-school children tonal audiometry is not possible. Though subjective, tonal audiometry is the most reliable method for evaluation of hearing threshold. Therefore, in children of this age, hearing threshold is evaluated by objective electroacoustic methods: early brainstem auditory evoked potentials (BAEP) and acoustic stapedius reflex. MATERIAL AND METHODS: In 60 adult examinees (120 ears) with normal hearing we analyzed the correlation between biological hearing threshold determined by tonal audiometry and threshold of ipsilateral stapedius reflex, as well as response threshold determined by brainstem evoked response audiometry (BERA). RESULTS AND DISCUSSION: We found a statistically significant correlation between biological hearing threshold and response threshold evaluated by brainstem evoked response audiometry. The linear Pearson correlation coefficient of: r = 0.40 for 500 Hz, r = 0.41 for 1,000 Hz, r = 0.44 for 2,000 Hz and r = 0.43 for 4,000 Hz (p < 0.01 for all values). Correlation coefficients between the biological hearing threshold and ipsilateral stapedius reflex threshold were not significant for 500 Hz (r = 0.46) and 1,000 Hz (r = 0.082), negative correlation was found for 2,000 Hz (r = 0.2656, p < 0.05) and significant correlation was confirmed only for 4,000 Hz (r = 0.225, p < 0.05). CONCLUSION: In regard to evident reliability of objective evaluation of hearing threshold by brainstem evoked response audiometry, this paper also suggests correction factors for evaluation of biological hearing threshold, and lists the deficiencies of hearing threshold assessment by this--still most accurate electrophysiological method. PMID- 11905182 TI - Clinical efficacy of goserelin (Zoladex) in the treatment of uterine myomas in infertile patients. AB - This study investigated the efficacy of Zoladex depot 3.6 mg (goserelin acetate) during a 4-month treatment of infertile patients with uterine myomas of different size and location. The investigation comprised 30 patients aged 22-42 years, distributed into 2 groups regarding uterine and myoma volume. The first group included patients with uterine myomas less than 70 ml and uterus less than 300 ml. The second group included patients in whom these volumes exceeded the above mentioned values. Zoladex depot was administered every 28 days for 4 months with ultrasonographic follow-up of volume decrease, whereas patients with submucous myomas underwent control hysteroscopy. The obtained results point to efficacy of Zoladex in decreasing the volumes of both myomas and uterus by more than 50%, which correlates with literature data. Of particular interest is complete disappearance of myomas in about 60% of patients of the first group. Serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) were followed-up prior to and during Zoladex therapy where multivariate variance analysis showed statistically significant differences. The side effects were recorded and are similar to those of other GnRH analogues. PMID- 11905183 TI - [From a scientific idea to clinical use]. AB - INTRODUCTION: In the nineteenth century, medicine, a former empiric skill, obtained its scientific basis owing to the development of physics, chemistry, biology, physiology, histology, biochemistry and other natural sciences. Fast progress in medicine was thus possible, but at the same time made the introduction of new scientific ideas into clinical use very difficult and therefore majority of them never reach satisfactory clinical application. Two examples from the author's own experience are presented here. FROM RADIOACTIVITY TO NUCLEAR MEDICINE: Soon after the discovery of radioactivity, George Hevesy gave a scientific basis for its use as a diagnostic and therapeutic tool in medicine. The first nuclear medicine laboratory was established in Vojvodina in 1963 and during the following twenty years, clinical application of radioactive tracers was developed very successfully in this area. But in the subsequent years the use of nuclear medicine methods declined dramatically for many reasons, mainly because of unsatisfactory cost/benefit ratio. FROM EXPERIMENTAL BLEEDING TO ERYTHROPOIETIN: The scientific idea of humoral regulation of hematopoiesis was proposed by Carnot at the beginning of the twentieth century, but it was introduced for clinical application only a hundred years later. It became possible by pharmaceutical production of recombinant hematopoietic growth factors by genetic engineering, and some of them, erythropoietin and granulocyte CSF, are available at high prices. They were used successfully for anemia in renal failure and some cases of neutropenic syndrome respectively, but compared to the initial expectations and quantity of research invested this is an unsatisfactory result. INSTEAD OF CONCLUSION: The introduction of these two presented scientific ideas into clinical application failed because of inadequate organization of medical scientific research and lack of financial support. PMID- 11905184 TI - [Clinical and diagnostic approaches to neurocysticercosis]. AB - ETIOLOGY: Cysticercosis is a tissue infection caused by larvae of the cestode Taenia solium. Neurocysticercosis is a central nervous system form of this infection. Taenia solium invades tissues in a form of a cyst with a thin, semitransparent wall. It can reach 1-2 cm in diameter in muscles and brain tissue and up to 3-6 cm in brain chambers. EPIDEMIOLOGY: Pigs are the most common intermediate hosts. The infection occurs when the parasite eggs or proglottids are ingested. It is most common in regions where human feces is used as a fertilizer, or regions with poor sanitary conditions. The man is infected with contaminated food or water, or by autoinoculation. PATHOGENESIS: The Taenia solium eggs are hatched in the duodenum. Embryos invade the intestinal mucosa, and reach various parts of the body, disseminated by the blood circulatory system. The most common localizations of cysticerci are skeleton, muscles and brain. While alive, these cysts produce a minimal reaction in hosts. Inflammation occurs when they die, often a few years after infection. CLINICAL FEATURES: While presence of adult worms of Taenia solium in the gastrointestinal tract causes unspecific symptoms, clinical features of neurocysticercosis depend on the number, size and localization of cysts, as well as on the degree of granulomatous response. The most common manifestations of this infection are epileptic seizures, whereas intracranial pressure increase can be the earliest sign of the disease. Hydrocephalus, meningitis and spinal cord compression syndrome are the most usual complications. PROGNOSIS: The death rate is low in neurocysticercosis with parenchymal cysts and calcification without hydrocephalus. However, fatal outcome occurs in hydrocephalic patients, cases with huge supratentorial cysts, multiple granuloma, brain edema or cerebral infarctions. DIAGNOSIS: Informations about travels to endemic regions are valuable in diagnosing neurocysticercosis. Cytobiochemical finding of the cerebrospinal fluid is often normal. However, in 50% of patients, lymphocytic or eosinophilic pleocytosis is found, low glucose (in 25%) and elevated protein (in 40% of cases). Further testing includes serologic examination of blood and cerebrospinal fluid. Finding of specific antibodies in the sera or cerebrospinal fluid confirms the diagnosis, although false positive reaction may occur in patients with other helminths, especially other cestode. The enzyme-linked immunotransfer blot assay is proven to be sensitive and specific in patients with multiple cysticerci. Computerized tomography and magnetic resonance imagining are techniques mostly used in establishing neurocysticercosis. They reveal the localization of cysticerci, identify the atrophic or edematous fields and assess the degree of ventricular dilatation. PMID- 11905185 TI - [Thrombocyte and leukocyte antigens]. AB - INTRODUCTION: By analogy, existence of erythrocyte antigens, classified in certain blood group systems, presence of specific alloantigens on platelets and leukocytes has been confirmed. CLINICAL SIGNIFICANCE OF PLATELET AND GRANULOCYTE ANTIGENS: Platelet and leukocyte antigens have a complex clinical significance, especially in hematology and blood transfusion. Immune response occurring after allosensibilization with platelet and granulocyte antigens is involved in pathogenesis of several clinical syndromes including: neonatal alloimmune thrombocytopenia, post-transfusion purpura, refractoriness to platelet transfusion, neonatal alloimmune neutropenia, transfusion related acute lung injury and chronic benign neutropenia in children. METHODS OF DETECTION: Application of various serological and molecular techniques enables phenotypization of platelet and granulocyte antigens and genomic analysis of DNA coding regions, providing determination of specific platelet and granulocyte alloantigen polymorphism. It confirms antigenic diversity of formed blood products. PMID- 11905186 TI - [Menopause in women in Novi Sad]. AB - INTRODUCTION: This study deals with the occurrence of menopause in women of Novi Sad during 1996 and 1997 in ambulatory-care services for women of the Health Center Novi Sad. MATERIAL AND METHODS: This retrospective study included 640 women, 42-82 years of age. RESULTS AND DISCUSSION: The mean age at menopause is 49.05 +/- 3.51 years, and median 50.05 +/- 0.17. Menopause in women under the age of 40 has been referred to as premature and it occurred in 16 women (2.49%). In regard to a longer time interval, in 585 women (91.41%) menopause occurred between 45-55 years of age. In our previous study of menarche in Novi Sad, we found that the mean age was 13.07 +/- 1.1 whereas the average reproductive period lasted 35.98 years. CONCLUSION: In order to perform relevant comparison of data in regard to menopause, it is necessary to establish a unified methodology. Foundation of Counseling Centers for menopausal women is desirable in the aim of better understanding their numerous problems during premenopausal and postmenopausal periods. PMID- 11905187 TI - [Salmonella bacteremia]. AB - INTRODUCTION: Non-typhi salmonellae are invasive bacteria, which can, in certain conditions, get into the blood stream, and course bacteriemia or localized infections in different organ systems. Incidence of salmonella bacteriemia varied in different studies between 5.2-13.7%. The incidence is higher in patients younger than 1 year and the elderly. Among different serotypes, the most invasive are Salmonella typhimurium, Salmonella choleraesuis and Salmonella Virchow. The aim of our study was to determine the incidence of salmonella bacteriemia among patients hospitalized at the Clinic of Infectious Diseases Novi Sad during 1991 1998, and to point to certain risk factors for salmonella bacteriemia. RESULTS: During 1991-1998, 1309 patients were hospitalized with the diagnosis of Salmonella gastroenteritis. 12 patients (0.99%) had positive blood culture. The average age of salmonella bacteriemia patients was 17 (1-54 year), but 50% of them were younger than 10. 4/12 patients (33.33%) had positive stool and blood culture, but in 8/12 (66.66%) only positive blood culture had been established. The main serotype was Salmonella enteritidis (83.33%) and after that Salmonella Virchow (16.66%). In more than 50% of patients there was at least one risk factor responsible for dissemination: sideropenic anemia (25%), cerebral palsy (8.33%) and chronic cardiac disease (8.33%). Average duration of fever was 13.16 days and diarrhea 8.83. All patients presented with hepatosplenomegaly. One patient had a localized salmonella infection (periappendicular abscess) and she had undergone successful surgery. All our patients had been under cephalosporins of the third generation for 2 weeks and they recovered completely without sequelae. CONCLUSIONS: A relatively low incidence of salmonella bacteriemia (0.99%) in our patients can be explained with small number of children younger than 1 year and patients older than 70 years of age in whom bacteriemia is most common. Positive blood culture doesn't mean obligatory positive stool culture. Routine laboratory analyses are relatively nonspecific for the diagnosis of generalized infection. Cephalosporins of the third generation are considered to be the drug of choice in treatment of salmonella bacteriemia. PMID- 11905188 TI - Autopsy material analysis at the Cardiology Clinic in 1999. AB - Clinico-pathological studies serve as a valuable source of information in everyday practice of most medical institutions. The aim of this study was to correlate the clinical and pathological diagnoses of principal disease and cause of death after autopsy in patients who died during 1999 at the Cardiology Clinic, Institute of Cardiovascular Diseases, Sremska Kamenica, Yugoslavia. Medical histories and autopsy reports of dead patients were analyzed: 255 (8.46%) patients died, but only 72 (28%) underwent autopsy. In 53 patients (73%) clinical cause of death was cardiac. Among these patients, consequences of coronary (atherosclerotic) artery disease prevailed in 41 patients (77%). Noncardiac cause of death was established in 19 patients (27%) and vascular causes prevailed with 17 (23%). The comparison of clinical and principal disease established by autopsy revealed a complete concordance in 62 cases, partial concordance in 1 and no concordance in 9 cases. Thus, concordance was found in 86-87.5%. The correlation coefficient was 0.48. Comparison of clinical and autopsy diagnoses of causes of death revealed concordance in 53 patients, with correlation coefficient 0.26. PMID- 11905189 TI - [Darier's disease--a familial case report]. AB - INTRODUCTION: Darier's disease is a slowly progressive autosomal dominant disorder characterized by a gene with variable penetrance. However, many cases of this disease are considered to be a new mutation in the genealogic tree. The prevalence of the disease was estimated as 1/55,000 to 1/100,000. CASE REPORT: The authors report a case of a female patient, 66 years of age, with Darier's disease, hospitalized at the Clinic of Dermatovenereology of the Clinical Center Novi Sad. The first changes of the skin occurred at the age of 16 originally on the shins and face, as miniature hard papules confluent in larger areas. Skin changes always exacerbated in the summer. At hospital admission the patient's head and hair were whole covered with white, thick, keratotics layer, resembling a helmet. The skin of the trunk was covered with white-gray, hyperkeratotic, fused papules like verrucous plaques, more expressed on the back. The skin of forearms presented with hyperkeratotic papules, and dorsum of the hands presented with plaques. Lower legs were covered with fused papilokeratotic, rough cauliflower-like layers with macerates and foetor. Buccal mucous was covered with whitish papules on erythematous lesions of cobblestone-like appearance. The nails were thickened with longitudinal furrowing, whereas the left third finger presented with V-shaped onychorrhexis. 4 generations were investigated, and the disease occurred only in the patient's younger daughter. DISCUSSION: Beside the classic "seborrheic" forms of Darier's disease there are a few clinical types: hypertrophied (intertriginous), vesiculo-bullous and linear (zosteriform) type. CONCLUSION: This case is a very severe classic form of Darier's disease, with variable penetrance and severity in the family. PMID- 11905191 TI - [Pheochromocytoma associated with cholelithiasis]. AB - OBJECTIVE: Pheochromocytoma is an adrenal or extra-adrenal tumor, which produces and secretes catecholamines. It is a rare cause of hypertension. Hypertension or hypertensive crises are most frequent clinical manifestations. Sometimes it can be associated with other diseases and conditions, such as cholelithiasis. CASE REPORT: We present a 37-year-old woman admitted to hospital with gastrointestinal complaints (right upper abdominal pain, nausea, vomiting) and hypertension with occasional hypertensive crises. Routine abdominal ultrasound revealed a gall bladder stone, and enlargement of the right adrenal gland. The diagnosis of pheochromocytoma was confirmed by measurement of catecholamine levels in 24-hour urine collection and MIBG scan. After preoperative preparation, tumor extirpation was done. Histology confirmed the diagnosis of pheochromocytoma. Routine controls of catecholamine 24-hour urine levels showed no catecholamine excess, without hypertension and hypertensive crises as most frequent clinical manifestations. COMMENT: Physicians must consider pheochromocytomas in cases of drug resistant hypertension or hypertensive crises. Sometimes pheochromocytomas are associated with other diseases, such as gall-bladder stones, with mimicking and overlapping of clinical manifestations of pheochromocytomas with symptoms and signs of the associated disease. PMID- 11905190 TI - [The superior vena cava syndrome as a manifestation of dissection of the ascending aorta]. AB - INTRODUCTION: Most clinical manifestations of aortic dissection are due to complications of either ischemic origin or wall rupture of pleural, pericardial, peritoneal or mediastinal cavity. Compression of other blood vessels such as pulmonary artery or superior vena cava is possible, but rarely occurs. CASE REPORT: A 60 year-old patient was admitted to hospital due to severe cyanosis and edema of the face, neck and upper thorax. Ten years ago, due to aortic insufficiency, aortic valve replacement with mechanical prosthesis (St. Jude) was performed. Diagnosis of superior vena cava syndrome was established on the basis of clinical examination, ECG and chest radiography. The etiology was confirmed by echocardiography indicating an enormous dissecting aneurysm of the ascending aorta, 9.2 cm in diameter. Lethal outcome followed 24 h after admission according to the type of electromechanical dissociation. DISCUSSION: The first case of superior vena cava syndrome was described by William Hunter in 1757. This severe disease is caused by tumors which compress or develop inside superior vena cava. In cases of rapid symptom occurrence, thrombosis or compression of vena due to hematoma (trauma, voluminous, dissecting aortic aneurysm) should be considered. Since symptoms of aortic dissection were absent (thoracic pain, aortic regurgitation, pulse asymmetry) the etiologic diagnosis of superior vena cava syndrome was confirmed by echocardiography. Surgical treatment of dissection provides repermeabilization of the superior vena cava and loss of symptoms. CONCLUSION: Superior vena cava syndrome is a rare and slightly known clinical manifestation of ascending aortic dissection. If symptoms rapidly occur, dissection should be considered, particularly in previously surgically treated patients. PMID- 11905192 TI - [131-I-lipiodol in therapy of liver carcinoma--methods and case report]. AB - INTRODUCTION: Primary malignant liver tumors are successfully treated only by means of surgery, but no more than 10% of patients are candidates for surgical intervention. The rest receive only palliative treatment which is, as a rule, unsuccessful. 131I-Lipiodol therapy (commercial label LIPIOCISTM) applied to the hepatic artery through a catheter has been used since 1984, primarily for treatment of hepatocellular carcinoma (HCC). The aim of this paper is to review the methodology of 131J-Lipiodol treatment of primary liver carcinomas. This treatment was applied for the first time in Yugoslavia. CASE REPORT: A female patient, 46 years of age had an inoperable primary liver carcinoma. Since progression of the disease couldn't be controlled by chemotherapy, treatment with 131I-Lipiodol was indicated. After blocking the thyroid with Lugol, Sol. 2.22 GBq of 131I-Lipiodol was injected into hepatic artery via catheter, and the patient was isolated in a designated facility until discharged. Around 5 months later, second therapeutic dose of 1.11 GBq was administered. Early post-therapy complications were severe, but transient. After 131I-Lipiodol therapy, the tumor growth was stopped, but the patient's general condition slowly deteriorated. The patient died 7 months after receiving the first therapeutic dose. CONCLUSION: In the reported patient, 131I-Lipiodol therapy stopped the tumor growth within the liver and significantly prolonged survival compared to the expected, but no improvement in quality of life was achieved. This treatment methodology is very complex. Medical staff providing care for these patients is exposed to substantial irradiation. PMID- 11905193 TI - [Pharmacies and pharmacists in Senta from 1816-1966]. AB - INTRODUCTION: This paper reviews historical data on the beginnings of pharmacy and first pharmacies in the world, with special emphasis on pharmacies at the territory of Vojvodina, as well as laws regulating education of pharmacists in the 19th century and work of pharmacies of that time. OUR INVESTIGATIONS: The first data on pharmacies of Senta date back to 1816 when Jozef Samanic submitted an application to open a pharmacy in Senta. The first pharmacy in Senta was founded in 1818 and was called "Kod crnog orla" (At the Black Eagle), whereas Jozef Helbling was the first pharmacist. He sold his pharmacy to Jozef Diener who then sold it to Ferenc Hajsler in 1826. The Hajslers owned this pharmacy in Senta for four generations. They were prominent and respected citizens of Senta. The second pharmacy in Senta was called "Narodna apoteka" (The Public Pharmacy) and was opened by Menjhart Muci in 1883, but was rented several times till 1935 when Lajos Herman bought it. In 1894 Kalmar Cizer got a permission to open the third pharmacy in Senta, but a year later he sold it to Vojislav Jovanovic. In 1929 he sold it to Mor Glikstal. In 1911 the fourth pharmacy was opened by Dula Banfi and was called "Apoteka nada" (The Pharmacy of Hope). It was inherited by Boldizar Banfi, a prominent pharmacist in Senta. The fifth pharmacy was founded by Zvonimir Tolovic and was called "Kod svetog Antuna" (At Saint Antun's). In 1929 he sold it to Laslo Husag. The sixth pharmacy was opened in 1938 by Obrad Popov. After the end of World War II in 1945, the first Public Pharmacy was founded. After that five public pharmacies were founded and they merged into one Public pharmacy in 1963. In 1966 there was a referendum and The Public Pharmacy became a part of the Medical Center founded on January 1, 1966. At the end, names of all registered pharmacists who used to work in Senta from 1818 to 1966 are listed, as well as dates when they began and finished working in our town. PMID- 11905194 TI - Cilia formation in the adult cat brain after pargyline treatment. AB - The brains of four adult cats treated with pargyline (a nonhydrazide monoaminoxidase inhibitor) were examined at both the light and electron microscopic levels. Formation of typical mature cilia with the 9 + 2 pattern was observed in neural cells in the following areas: habenula nuclei, interpeduncular nuclei, hippocampus, mammillary bodies, thalamus, and caudate nucleus. The most marked ciliation occurs in the habenula nuclei. In general, glial cells greatly predominate in the formation of cilia. It is not clear whether ciliation in the central nervous system is the direct result of pargyline or if it occurs indirectly as a result of inhibition of monoaminoxidase. These findings are compared with the serotonin effect on ciliation in the embryogenesis of lower forms. It is suggested that pharmacological stimulation of centriolar reproduction without subsequent mitosis may lead to ciliary formation. PMID- 11905195 TI - The effect of prefixation on the diameter of chromosome fibers isolated by the Langmuir trough-critical point method. AB - The effect of prefixation on the diameter of chromosome fibers isolated by the Langmuir trough-critical point method has been investigated in several species of plants and animals. In barley, fibers isolated from endosperm without prefixation have an average diameter of between 240 and 250 A, and are similar in dimensions and structure to the chromosome fibers isolated from animals by this method. Chromosome fibers from other tissues of the same plant are smaller in diameter when isolated without prefixation, approximating 200 A. After prefixation in 2% buffered formalin, isolated fibers from the three barley tissues studied are reduced in diameter, to approximately 120-130 A for endosperm and leaflet and to 140 A for root tip. Chromosome fibers isolated from newt erythrocytes also show a significantly reduced diameter after formalin prefixation, to approximately 120 A. PMID- 11905196 TI - Structure of coupled and uncoupled cell junctions. AB - Cells of Chironomus salivary glands and Malpighian tubules have junctions of the "septate" kind. This is the only kind of junction discerned which is large enough to effect the existing degree of intercellular communication. The electron microscopic observations of the "septate" junction conform to a honeycomb structure, with 80-A-thick electron-opaque walls and 90-A-wide transparent cores, connecting the cellular surface membranes. A projection pattern of light and dark bands (the "septa") with a 150-A periodicity results when the electron beam is directed normal to any set of honeycomb walls. Treatment of the salivary gland cells with media, which interrupt cellular communication (without noticeable alteration of cellular adhesion) by reducing junctional membrane permeability or perijunctional insulation, produces no alterations in the junctional structure discernible in electron micrographs of glutaraldehyde-fixed cell material. PMID- 11905198 TI - Electrophysiological evidence for low-resistance intercellular junctions in the early chick embryo. AB - Electrophysiological evidence is presented for the exchange of small ions directly between cells interiors, i.e. "electrical coupling," in the early chick embryo. Experiments with intracellular marking show that coupling is widespread, occurring between cells in the same tissue, e.g. ectoderm, notochord, neural plate, mesoderm, and Hensen's node, and between cells in different tissues, e.g. notochord to neural plate, notochord to neural tube, notochord to mesoderm. The coupling demonstrates the presence of specialized low-resistance intercellular junctions as found in other embryos and numerous adult tissues. The results are discussed in relation to recent electron microscopical studies of intercellular junctions in the early chick embryo. The function of the electrical coupling in embryogenesis remains unknown, but some possibilities are considered. PMID- 11905197 TI - Structural modulations of plasmalemmal vesicles. AB - Structural modulations affecting a small fraction of the population of plasmalemmal vesicles of vascular endothelia are described. They include forms which are apparently produced by the fusion of the vesicular membrane with the plasmalemma and by the successive elimination of the layers of the two fused membranes. Such modulations are assumed to represent stages in the discharge process of vesicular contents. Other forms, characterized by their flask shape and elongated neck, are assumed to represent stages in the formation and loading of membrane invaginations, followed by their being pinched off to form isolated vesicles. Stages in a membrane-fusion process leading to the formation of apertured fenestrae and channels are also described in fenestrated endothelia. The visualization of these structural details is greatly facilitated by staining tissue specimens with uranyl acetate before dehydration. PMID- 11905199 TI - Effect of acriflavin on the kinetoplast of Leishmania tarentolae. Mode of action and physiological correlates of the loss of kinetoplast DNA. AB - The loss of kinetoplast DNA in Leishmania tarentolae, which occurs in the presence of low concentrations of acriflavin, was found to be a result of selective inhibition of replication of this DNA. Nuclear DNA synthesis was relatively unaffected and cell and kinetoplast division proceeded normally for several generations. An approximately equal distribution of parental kinetoplast DNA between daughter kinetoplasts resulted in a decrease in the average amount of DNA per kinetoplast. The final disappearance of the stainable kinetoplast DNA occurred at a cell division in which all the remaining visible kinetoplast DNA was retained by one of the daughter cells. The selective inhibition of kinetoplast DNA synthesis was caused by a selective localization of acriflavin in the kinetoplast. The apparent intracellular localization of dye and the extent of uptake at a low dye concentration could be manipulated, respectively, by varying the hemin (or protoporphyrin IX) concentration in the medium and by adding red blood cell extract (or hemoglobin). Hemin and protoporphyrin IX were found to form a complex with acriflavin. During growth in acriflavin, cells exhibited an increasing impairment of colony-forming ability and rate of respiration. No change in the electrophoretic pattern of total cell soluble proteins was apparent. The data fit the working hypothesis that the loss of kinetoplast DNA leads to a respiratory defect which then leads to a decrease in biosynthetic reactions and eventual cell death. A possible use of the selective localization of acriflavin in the kinetoplast to photooxidize selectively the kinetoplast DNA is suggested. PMID- 11905200 TI - The distribution of DNA among dividing mitochondria of Tetrahymena pyriformis. AB - A squash technique was developed for log phase Tetrahymena pyriformis which permitted the resolution of over 100 individual mitochondria from a single cell. Mitochondria incorporated thymidine at all stages of the cell cycle, even when nuclear DNA synthesis was not occurring. During the stage of macronuclear DNA synthesis, however, there was a significant increase in the extent of mitochondrial labeling. Low radioautograph background suggests that mitochondrial DNA is synthesized at the mitochondria themselves. All mitochondria incorporated thymidine-3H within one population-doubling time. Grain counts also showed that the amount of mitochondrial label was retained for four generations and that this label remained randomly distributed among all mitochondria during this time. The results are not consistent with any theory of de-novo or "microbody" origin of mitochondria, but do support the hypothesis that mitochondria are produced by the growth and division of preexisting mitochondria. The stability of the mitochondrial DNA and its distribution among daughter mitochondria satisfy two prerequisites for a genetic material. The possibility is discussed that some of the genetic information for the mitochondrion is contained in the DNA associated with this organelle. PMID- 11905201 TI - Functional consequences of ultrastructural geometry in "backwards" fluid transporting epithelia. AB - Many fluid-transporting epithelia possess dead-end, long, and narrow channels opening in the direction to which fluid is being transported (basal infoldings, lateral intercellular spaces, etc.). These channels have been thought to possess geometrical significance as standing-gradient flow systems, in which active solute transport into the channel makes the channel contents hypertonic and permits water-to-solute coupling. However, some secretory epithelia (choroid plexus, Malpighian tubule, rectal gland, etc.) have "backwards" channels opening in the direction from which fluid is being transported. It is shown that these backwards channels can function as standing-gradient flow systems in which solute transport out of the channel makes the channel contents hypotonic and results in coupled water flow into the channel mouth. The dependence of the transported osmolarity (isotonic or hypertonic) on channel radius, length, and other parameters is calculated for backwards channels for values of these parameters in the physiological range. In addition to backwards channels' being hypotonic rather than hypertonic, they are predicted to differ from "forwards" channels in that some restrictions are imposed by the problem of solute exhaustion, and in the presence of a sweeping-in effect on other solutes which limits the solutes that may be transported. PMID- 11905202 TI - Development of the metanephric kidney. Protein and nucleic acid synthesis. AB - The metanephric kidney was studied in fetal and older mice beginning at 16 days after mating of the parents. Polyribosomes from fetal kidneys labeled in vitro with 14C-labeled amino acids had 10-20 times more acid-precipitable radioactivity associated with them than polysomes from adult kidneys similarly labeled. Between 3 and 6 days after birth the rate incorporation of labeled amino acids by polyribosomes from neonatal kidneys declined sharply to only twice the value found for adult kidneys. There was no change in the shape of the polyribosome profile with increasing age, but before birth few, if any, ribosomes were bound to membranes compared with 20% 2 days after birth and between 20 and 30% in the adult. Total protein represented less than 10% of the wet weight in the fetal kidney but increased to 17% of the wet weight in the adult kidney. There was a steady decline in the concentration of RNA and DNA with respect to dry weight throughout kidney development. DNA concentration declined more rapidly than RNA concentration, so that the milligram to milligram ratio of RNA to DNA increased. In males the RNA/DNA ratio was stable at 1.3 at 40 days after birth; but in females the decline in DNA concentration was more protracted, and at 200 days after birth the RNA/DNA ratio was only 0.99. Thus, total nucleic acids show only gradual changes in concentration throughout development of the kidney, but a sharp change in the synthetic activity of the ribosomes and in their binding to membranes occurs in kidneys soon after birth. PMID- 11905203 TI - Pinocytotic response of circulating erythrocytes to specific blood grouping antibodies. AB - Human blood samples from adults and newborns of blood groups O, A, and B were treated with either anti-A blood grouping serum, ferritin-conjugated anti-A serum, free ferritin, or saline and then prepared for electron microscopy. Morphological differences were observed between the untreated erythrocytes of infants and adults. Circulating red cells of newborns were frequently vesiculated (25.5%), whereas those of adults only occasionally showed vesicles (5.5%). On the basis of morphology and incidence, the majority of these vesiculated cells seemed to be mature erythrocytes. The introduction of anti-A serum to group A erythrocytes of infants appeared to stimulate vesicle formation, but anti-A serum did not have a similar effect on group O or B cells of infants or on group A cells of adults. Vesicles which formed in response to antiserum treatment appeared to be the result of pinocytosis. In contrast to the well dispersed ferritin along the membrane of agglutinated adult cells, the ferritin particles on the infants' cells were frequently clustered at irregular intervals. These accumulations seemed to lead to invaginations of the cell membrane, resulting in ferritin-lined intracytoplasmic vesicles. The addition of free ferritin or ferritin-conjugated antibodies of the wrong specificity to red cells did not increase vesicle formation. PMID- 11905204 TI - Membranes of animal cells. II. The metabolism and turnover of the surface membrane. AB - Turnover studies of the surface membrane and of cell particulate matter of L cells in tissue culture in logarithmic and plateau phase of growth have been made. The rate of incorporation of isotope into these fractions and the rate of fall of specific activities of labeled L-cell fractions have been observed. The following interpretation of the data appears most likely although other interpretations are possible. Growing and nongrowing cells synthesize approximately similar amounts of surface membrane and particulate material. In the growing cell the material is incorporated with net increases in substance. There is relatively little turnover. In the nongrowing cell newly synthesized material is incorporated, but a corresponding amount of material is eliminated so that there is turnover without net increase of substance. Our results suggest that there is no gross differential turnover between the protein, lipid, and carbohydrate of the surface membrane under the conditions of our experiments. Metabolic inhibitors or omission of amino acids in the culture medium lead to a decrease in synthesis of surface membrane and cell particulates and cause an equivalent decrease in the rate of degradation of surface membrane and of particulates; therefore the synthetic and degradative aspects of turnover appear to be coupled. As cultures of nongrowing cells in suspension or on a glass surface age, their synthetic and turnover capacities diminish. Our results suggest that the cell may exist in a nongrowing state with a level of synthesis similar to that of a growing cell. It can exist in this state with a high level of turnover. PMID- 11905206 TI - Regulation of DNA replication in the nuclei of the slime mold Physarum polycephalum. Transplantation of nuclei by plasmodial coalescence. AB - Nuclei in G2 phase of the slime mold Physarum polycephalum, when transplanted, by plasmodial coalescence, into an S-phase plasmodium, failed to start another round of DNA synthesis. In the reciprocal combination, S-phase nuclei in a G2-phase host continued DNA synthesis for several hours without appreciable decrease in rate. It is suggested that the beginning of DNA replication is determined by an event, either during or shortly after mitosis, which renders the chromosomes structurally competent for DNA replication. PMID- 11905207 TI - Control of cell progression through the mitotic cycle by carbohydrate provision. I. Regulation of cell division in excised plant tissue. AB - A stationary phase in the root meristem of excised pea roots was established by prolonged carbohydrate deprivation in sterile culture medium. When the stationary phase had been established, cells that had collected in the G1 period of the mitotic cycle were induced to enter the S stage by subjection to relatively short intervals of carbohydrate provision (sucrose spurts). Progression and cycle location of the G1 cells induced to enter S were measured with tritiated thymidine and radioautography. The results indicated that the number of G1 cells induced to enter S increased directly with the spurt duration and that cells could be positioned and retained in the S and/or G2 periods by varying the duration of the spurt. The data support the hypothesis that S and maybe M stages have a relatively larger dependence on carbohydrate availability, and presumably a greater energy requirement, than G1 and G2. PMID- 11905205 TI - Studies on the permeability of calf thymus nuclei isolated in sucrose. AB - A study of the permeability of calf thymus nuclei isolated in sucrose was carried out with sucrose-14C, glycerol-14C, and carboxydextran-14C (molecular weight, 60,000-90,000). The results indicate that the nuclei are very permeable to both sucrose and glycerol but they exclude the carboxydextran. Results obtained with other low molecular weight non-electrolytes (malonamide-14C, erythritol-14C, D arabinose-14C, and D-mannitol-14C) are in agreement with the view that the nuclei are freely permeable to these molecular species. A sucrose-impermeable space is also present in these preparations and it has been attributed to the presence of intact cells. The high permeability of nuclei to sucrose was confirmed with Ficoll-separated preparations. The possibility of the presence of a substantial particulate space that allows the penetration of dextran cannot be excluded by these experiments, and this space may correspond to damaged nuclei. PMID- 11905208 TI - The ultrastructural basis of alveolar-capillary membrane permeability to peroxidase used as a tracer. AB - The permeability of the alveolar-capillary membrane to a small molecular weight protein, horseradish peroxidase (HRP), was investigated by means of ultrastructural cytochemistry. Mice were injected intravenously with HRP and sacrificed at varying intervals. Experiments with intranasally instilled HRP were also carried out. The tissue was fixed in formaldehyde-glutaraldehyde fixative. Frozen sections were cut, incubated in Graham and Karnovsky's medium for demonstrating HRP activity, postfixed in OsO4, and processed for electron microscopy. 90 sec after injection, HRP had passed through endothelial junctions into underlying basement membranes, but was stopped from entering the alveolar space by zonulae occludentes between epithelial cells. HRP was demonstrated in pinocytotic vesicles of both endothelial and epithelial cells, but the role of these vesicles in net protein transport appeared to be minimal. Intranasally instilled HRP was similarly prevented from permeating the underlying basement membrane by epithelial zonulae occludentes. Pulmonary endothelial intercellular clefts stained with uranyl acetate appeared to contain maculae occludentes rather than zonulae occludentes. HRP did not alter the ultrastructure of these junctions. PMID- 11905209 TI - The nervous environment of individual smooth muscle cells of the guinea pig vas deferens. AB - Smooth muscle cells of the external longitudinal coat of the guinea pig vas deferens were followed for 480 mu at 4.5-mu intervals. Muscle bundles and fibers interwove, facilitating intermuscular and neuromuscular contacts. The ribbon- or rodlike muscle cells were about 450 mu long, 3,000 mu3 in volume, and 4,500 mu2 in area. The thickened nuclear zone day anywhere along the middle one-third of the cell. Intercellular distances were 500-800 A. Intrusions were rare, and tight junctions absent. At any level in a field of 80 muscle fibers there were 10-15 nerve bundles, each containing several varicose axons. Bundles and axons divided. Axons, en passage, were frequently within 500-1,000 A of a muscle fiber. En passage close contacts were rate. Axon terminations were bare, and bare axons invariably terminated. Bare terminations had scattered vesicle-laden varicosities and were from 10-60 mu in length, and all ended within 500 A of muscle fibers. Some made close contact with muscle fibers. Less than half of the muscle cells received this close contact, but some cells were approached by more than one termination. Most terminations involved more than one cell. Some cells had little or no innervation. Some groups of cells had a rich innervation. There was very little evidence of sensory innervation. These conclusions are not valid for other smooth muscles. PMID- 11905210 TI - Unique cytoplasmic membranes in Rous sarcoma virus-induced tumors of a subhuman primate. PMID- 11905211 TI - Protein: lipid ratios of liver mitochondria during development. PMID- 11905212 TI - A reinvestigation of cross-sections of cilia. PMID- 11905213 TI - The presence of a crystalline matrix in pyrenoids of the diatom, Achnanthes brevipes. PMID- 11905215 TI - Specialized "transfer cells" in minor veins of leaves and their possible significance in phloem translocation. PMID- 11905214 TI - Behavior of the gamete membranes during sperm entry into the mammalian egg. PMID- 11905216 TI - The woman with a candle. Finding scholars of pediatric nursing. PMID- 11905217 TI - Asthma self-management programs for children. Part II: Empiric evaluation of programs and recommendations. PMID- 11905218 TI - Reducing children's immunization distress in a primary care setting. PMID- 11905220 TI - Home visits. Should they remain a significant component of today's pediatric healthcare continuum? PMID- 11905219 TI - Factors associated with significant injuries in youth ice hockey players. PMID- 11905221 TI - Assessment of skin breakdown risk in children. PMID- 11905222 TI - Evaluating the validity of Internet sources. PMID- 11905223 TI - Family focus. PMID- 11905224 TI - [Prevalence of weather sensitivity in Germany]. AB - BACKGROUND AND OBJECTIVE: As epidemiological data on prevalence of weather related health effects were lacking we conducted a weather sensitivity (WS) survey in Germany. SUBJECTS AND METHODS: A total of 1064 citizens (age > 16) representative of the population in Germany were interviewed in January 2001. RESULTS: The results show that 19.2% of the population believe that weather influences their health to a "high degree", 35.3% that weather has "some influence on their health". The highest prevalence of WS (high + some influence) is found in the age group of > 60 years, in 68% of the subjects. The highest frequencies of weather-related symptoms are reported for stormy weather (30%) and when it gets colder (29%). The most frequent symptoms reported by weather sensitive subjects are headaches/migraine (61%), lethargy (47%), sleep disturbances (46%), fatigue (42%), joint pain (40%), irritation (31%), depression (27%), vertigo (26%), concentration problems (26%) and scar pain (23%). 32% of the weather-sensitive subjects have been incapable to do their regular work because of weather-related symptoms at least once last year, 22% even several times. Co-morbidity was significantly higher in weather-sensitive subjects. CONSEQUENCE: On the basis of these data we are planning studies on causal factors of weather-related health effects. PMID- 11905225 TI - [Histoplasmosis group disease in bat researchers returning from Cuba]. AB - CASE HISTORY: Four males (age 25 to 40 years) and one female (age 25) were admitted to our hospital almost simultaneously with symptoms of fever up to 38 degrees C, dry cough, thoracodynia, dyspnoea, myalgia and arthralgia. All patients belonged to a team of eight German bat researchers who had returned from Cuba 10 days before, where they had investigated bats in caves. Another member of the team had only mild histoplasmosis and was followed in our outpatient clinic. Two scientists who wore their breathing masks continuously during their work in the caves did not fall ill. EXAMINATIONS: Chest X-rays of all in-patients showed pulmonary infiltrates correlating with the severity of their illness. In all patients specific IgG antibodies against Histoplasma capsulatum-antigen were found in the Western Blot assay, confirming the diagnosis of histoplasmosis. TREATMENT: Treatment with oral itraconazole 200 mg b.d was given to four inpatients for 6 weeks, in the fifth patient itraconazole was discontinued because of an increase of liver transaminases. CONCLUSION: Antimycotic treatment of advancing histoplasmosis seems appropriate also in immunocompetent patients. The high number of patients within this group suggests high numbers of Histoplasma capsulatum in the caves. Wearing breathing masks throughout the work in the caves may prevent histoplasmosis even in case of high infectious doses. Pre-travel recommendations for cave researchers have to emphasize the continuous use of breathing masks and vaccination against tetanus and rabies. PMID- 11905226 TI - [A hemodynamically active type II atrial septal defect in a 78-year-old patient. Indications for interventional catheter occlusion?]. AB - HISTORY AND ADMISSION FINDINGS: A 78-year-old woman presented with a first episode of syncope. She reported increasing fatigue and dyspnoea upon exertion over a period of 20 years and chest pain 2 months prior to admission. Auscultation revealed fixed doubling of the second heart sound. INVESTIGATIONS: Laboratory tests showed increased troponin I. Transaminases were moderately elevated. Chest X-ray showed an enlarged right heart and a dilated pulmonary artery (2 cm). Echocardiography discovered a large secundum atrial septal defect with a diameter of 3 cm but no right to left shunt (no Eisenmenger reaction). Cardiac catheterization revealed a stenosis of the right coronary artery and severe systolic pulmonary hypertension of 80 mmHg. DIAGNOSIS, TREATMENT AND COURSE: A significant stenosis of the right coronary artery was successfully dilated. The ASD was closed by interventional implantation of a commercial closure device (Amplatzer). One month later, echocardiography indicated in an estimated systolic pulmonary pressure of 30 mmHg. The patient's condition improved considerably. CONCLUSION: This case is remarkable in that a very large ASD was asymptomatic up into old age and without the development of an Eisenmenger reaction. Also, large ASD can be by catheterization with the appropriate closure device. Fixed pulmonary hypertension is not obligatory. Non invasive closure is a good alternative of surgery in elderly patients with risk factors. PMID- 11905227 TI - [Diagnosis of acute renal failure]. PMID- 11905228 TI - [Is public health facing a paradigm change?]. PMID- 11905229 TI - [Right-sided sweating of the head in arterial hypertension]. PMID- 11905230 TI - [Vague thoracic symptoms]. PMID- 11905231 TI - [What is the value of measuring 2-hour blood sedimentation rate?]. PMID- 11905232 TI - [Contrast medium-induced renal failure can not be prevented by hemodialysis]. PMID- 11905233 TI - [Acute intestinal hemorrhage in marked diverticulosis of the small intestine]. PMID- 11905234 TI - [Simultaneous surgical treatment of internal carotid artery stenoses and coronary heart disease]. PMID- 11905235 TI - [Cystic ovarian disease in dairy cattle: aetiology, pathogenesis, and risk factors]. AB - Cystic ovarian disease (COD) is an important ovarian dysfunction in dairy cattle, especially during the early postpartum period. The endocrinology and the symptoms of this disorder vary widely due to the many different forms of cysts that exist. For these reasons, there is currently no clear and unambiguous definition of COD. When ovulation does not occur, a follicle may evolve into an ovarian cyst. Folliculogenesis and ovulation are regulated by the hypothalamic-pituitary gonadal axis. A dysfunction can occur at different levels of this neuroendocrine system, causing COD. The primary factor is thought to be a deficient luteinizing hormone surge prior to ovulation. What causes this alteration is not yet known. Many factors increase the incidence of COD and are involved in the very complex pathogenesis of this disease. PMID- 11905236 TI - [Can gene technology in agriculture prevent hunger in the world?]. AB - The world population grows rapidly: the number of mouths to feed increases. Is an agriculture without gene technology able to produce sufficiently in order to prevent hunger? Research indicates that hunger is not the result of short comings in agricultural outputs. It is however the result of poverty. This problem will not be solved by gene technology based agricultural production. This article explains the basic principles of mainstream and organic farming. Literature shows that the production potentials of both kinds of farming are, by far most, not yet exhausted. Gene technology is therefore unnecessary. PMID- 11905237 TI - [Fever in the cat]. PMID- 11905238 TI - [Udder health panel speaks out about advising for practices and risk factors for udder infections]. PMID- 11905239 TI - [No, I don't want to!]. PMID- 11905240 TI - [The contagiousness of the pestivirus and foot and mouth disease virus]. PMID- 11905241 TI - [Defense re-hires veterinary specialists]. PMID- 11905242 TI - [Reaction to the letter of A. van der Paauw]. PMID- 11905243 TI - [BSE and species specificity]. PMID- 11905244 TI - Adhesion molecules of immunoglobulin gene superfamily in stroke. AB - Stroke-induced inflammatory reaction leads to the accumulation of leukocytes in the brain ischaemic region, where they exert a detrimental effect--promotion and extension of cerebral damage. Intracerebral infiltration of peripheral blood leukocytes requires prior endothelial-leukocyte interactions that are mediated by such cell surface proteins as adhesion molecules. Among adhesion molecules, it is the immunoglobulin gene superfamily (IgSF) that is responsible for strong attachment and transendothelial migration of leukocytes. The principal members of IgSF are: intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1). In this review the following issues were described and discussed: an increased expression of ICAM-1 and VCAM-1 in ischaemic brain as well as a detection of their soluble(s) forms in sera of stroke victims. The presented data suggest the involvement of both ICAM-1 and VCAM-1 in the sequence and timing of the infiltration of leukocytes into the brain ischaemic zone after stroke. They have also revealed changes in serum concentrations of sICAM-1 and sVCAM-1 that are characteristic for stroke. Recently, increase in sPECAM-1 levels in serum and cerebrospinal fluid (CSF) has been shown within 24 h of the onset of stroke, having indirectly suggested involvement of the molecule in the inflammatory events during the early phase of stroke. PMID- 11905245 TI - Uterus-innervating neurones of paracervical ganglion in the pig: immunohistochemical characteristics. AB - Immunohistochemical characteristics of neurones innervating the porcine uterus located in paracervical ganglia were studied with a combination of retrograde fluorescent tracing and immunofluorescence. Retrograde fluorescent tracer Fast Blue (FB) was injected into the uterine horn and uterine cervix. The presence of biologically active substances, tyrosine hydroxylase (TH), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), galanin (GAL), Met-enkephalin-Arg-Gly Leu (MEAGL) and calcitonin gene-related peptide (CGRP) was studied in FB-positive neurones localised in paracervical ganglia. FB-positive neurones containing TH, NPY, VIP and MEAGL were numerous, while those containing CGRP were scarce. The results pointed to some species-related differences in immunohistochemical coding of neurones of paracervical ganglion responsible for uterus innervation. PMID- 11905246 TI - The morphological types of neurones of the medial and lateral mamillary nuclei in a newborn guinea pig: Nissl, Kluver-Barrera and Golgi studies. AB - The studies were carried out on the hypothalamus of 5 newborn (P0 stage) guinea pigs. The sections were impregnated according to three modifications of the Golgi technique or stained according to the Nissl and Kluver-Barrera methods. On the basis of the shape and size of perikarya, dendroarchitecture, pattern of axon as well as the inner structure of neurones, in the medial (Mm) and lateral (MI) mamillary nuclei four morphological types of nerve cells were distinguished: cap like with two subtypes (33% of the cell population), fusiform (35%), triangular (12%) and rounded unidendritic (21%) neurones. The majority of them possessed spines on their dendrites. The spiny cells, both cap-like and fusiform ones, were observed preponderantly, in the medial mamillary nucleus, whereas in the lateral mamillary nucleus there were mainly seen the triangular and fusiform neurones, either spiny or aspiny cells. The spineless rounded unidendritic cells were dispersed throughout the mamillary region, but they were twice as numerous in Mm as in MI, where they were the least numerous. PMID- 11905247 TI - Visualisation of diverticula of the upper part of the alimentary tract; comparison of roentgenologic and endoscopic techniques. AB - Diverticula of the upper part of the alimentary tract, irrespective of their etiology, are frequently observed benign changes of the pharynx, oesophagus, stomach and duodenum. In the present work, patients of the II General Surgery Department of the Medical University of Lublin, with radiologically or endoscopically proved diverticula of the upper part of the alimentary tract, were examined. The presence of diverticula of such localisation was an indication for supplementary endoscopic or radiological examination. The localisation, size, diameter of the opening, mucosal relief of diverticula and its contiguity were checked and analysed. Our data suggest that both medical procedures are complementary to each other. All previously observed changes in diverticula of the thoracic part of the oesophagus and the infradiaphragm part of the alimentary tract were fully proved. The radiological examination gave a better view of Zenker's diverticulum, especially in short and obese patients. Sampling and better visualisation of the diverticula opening testify to the unquestionable superiority of endoscopy. However, precise evaluation by radiological process fully completes the diagnostic protocol. Both diagnostic procedures are usually supplemented by manometric examination of the oesophagus and superior and inferior oesophageal sphincters. This enables the accurate diverticula etiology to be stated. PMID- 11905248 TI - Effect of unilateral, intraovarian infusions of bacteria on ovarian morphology in gilts. AB - The aim of this study was to investigate whether unilateral, intraovarian infusions of bacteria might have induced morphological changes in the contralateral ovary. Eleven sexually matured gilts with controlled estrous cycle were used. The animals were randomly divided into two groups: I (Gr. I, treated; n = 4), and II (Gr. II, control; n = 7). In Gr. I, 1 ml of bacterial suspension (10(3) colony forming units/ml of saline of Escherichia coli, Staphylococcus aureus and Corynebacterium pyogenes, in proportion 1:1:1) was infused into the hilus of one ovary from the 15th to the 19th day of the estrous cycle. At the same time, 1 ml of saline was infused into the hilus of the contralateral ovary and into both ovaries of the control gilts. On the 7th day of the next cycle, the ovaries were dissected out. There were no significant differences in the number of follicles and corpora lutea (CL) as well as in weight and size between the bacteria-infused, contralateral and control ovaries. The microscopic observations of the bacteria-infused ovaries revealed the presence of focal infiltrations of neutrophils in the softened stroma, especially around dilated blood vessels filled with erythrocytes. In the contralateral ovaries, the number of regularly distributed neutrophils in the softened stroma was greater than that found in the bacteria-treated ovaries. CL of the bacteria-infused ovaries had more numerous, dilated blood vessels than CL observed in the contralateral gonads. More neutrophils were found in CL of both ovaries in Gr. I as compared to those observed in Gr. II. In Gr. II, single neutrophils were found also in the stroma where the tip of the cannula was inserted. This study revealed that in gilts, unilateral, intraovarian administration of bacteria did not change the number of ovarian structures, the weight and size of the bacteria-infused and contralateral ovary, but induced inflammatory changes in both ovaries. PMID- 11905249 TI - Development of valves in the small saphenous vein in human fetuses. AB - The study was performed on 82 small saphenous veins in human fetuses of both sexes aged 9 to 37 weeks. The earliest valves were observed in 13 week-old fetuses. In fetuses between 13 and 18 weeks old the number of valves increased from 1 to 8. In the older fetuses between 19 and 37 weeks the number of valves varied from 5 to 9 and does not seem to be related to age. During development more valves were found in the upper part of the small saphenous vein. The height of valves increases with age and differs between particular valves of the same vein. PMID- 11905250 TI - Multiple vessel variations in the retropubic region. AB - We encountered some multiple vessel variations in the retropubic region of a 55 year-old male cadaver. The obturator artery had its origin from the external iliac artery, and inferior epigastric artery from the femoral artery. Additionally, an anastomosis between obturator and inferior epigastric veins (venous Crown of death) was observed. PMID- 11905251 TI - Bilio-pancreatic anomalies obscured with MRCP. AB - In this article the authors discuss whether or not diagnostic potential of MR cholangiopancreatography is strong enough to replace direct cholangiography in all cases. The pre-surgery analysis of a variety of pancreato-biliary disorders diagnosed using MRCP images is presented with the emphasising the importance of source images. Six cases of pancreato-biliary disorders are presented in which MRCP indicated the place of ductal stenosis as well as the morphologic variants or ductal uninspected shape which is critical for surgery or planned drainage. Coronal and axial MRCP source and MIP images were obtained with 0.5 T Gyroscan NT. Anomalies of the biliary or pancreatic ducts included two cases of choledochal cystic dilatation; two cases of aberrant biliary ducts, one case of gallbladder duct variant and a case of an additional pancreatic duct. In 3 out of 6 cases, the MRCP source images produced using the complementary method supplied more complete information concerning ductal junctions than the MIP images. Whereas in 3 out of 6 cases, both kinds of images were equally reliable. In 4 out of 6 cases, endoscopy was performed, and in 2 cases ERCP images were not diagnostic for ductal anatomy. However, full delineation of biliary and pancreatic ducts was complete in all MRCP images. MRCP within source images and maximum intensity projections show particular promise for the assessment of pancreato-biliary anomalies in order to reduce the number of higher-risk endoscopic interventions. The technique should be the method of choice in cases of suspected pancreato-biliary anomaly resulting from any imaging modality and is helpful for planning the optimal drainage method. In the long run this practice would reduce the number of ducts damaged during surgery. PMID- 11905252 TI - Aberrant left subclavian artery. AB - This paper describes a rare case in which the left subclavian artery originates from a common stem arising from the aortic arch and splits into a brachiocephalic trunk and a left subclavian artery. The course of other large vessels of the aortic arch in this case are typical. PMID- 11905253 TI - Neurological symptoms as the result of enlarged dimensions and non-typical course of inferior superficial temporal vein. AB - Neurological symptoms as the result of non-typical course of superficial cerebral veins are described in available literature very rarely. The case described below indicates that in some circumstances the compression symptoms derived from the cerebral cortex may be incredibly more serious than their anatomical reasons. In our observation a young woman was described complaining of paroxysmal numbness of the left upper limb with paraesthesia of the left side of the face, the left eye and left half of the tongue. The patient said that in childhood she used to have paroxysmal itching of the left hand. She also said that CT of the head made a few years ago after a car accident was without pathological changes. Neurological examination, x-ray of the skull and EEG test performed during first visit proved normal. After one year of the disease, Jackson-type epilepsy, combined with loss of sensation of the left half of the face for the first time, was present. Neurological and ophthalmological examination of the bottom of the eye proved normal. Skull x-ray was normal. Then disturbances of the vision in the left half of the field appeared. EEG was still in norm. The MRI test showed the asymmetry in the course and dilated superficial vein between the basis of the right temporal lobe and the tentorium of the cerebellum. The diameter of this vein was 2.5 mm, but there were no vascular malformations. Bottom of the eye was normal, but in the field of vision the white and red colours were dominated. PMID- 11905254 TI - Cholinergic innervation and calretinin-immunoreactive neurones in the hippocampus during postnatal development of the rat brain. AB - Immunohistochemical study of the cholinergic innervation of the hippocampal calretinin-containing cells was conducted on 28 rat brains of postnatal ages: P0, P4, P7, P14, P21, P30 and P60. Sections with double immunostaining for vesicular acetylcholine transporter (VAChT; the marker of cholinergic cells, fibres and terminals) and calretinin were analysed using confocal laser-scanning microscope. Obtained data demonstrate that during development as well as in adult species calretinin-containing neurones in the rat hippocampus form sparse synaptic contact with VAChT-ir terminals. It seems probable that cholinergic innervation is not crucial for the functioning of CR-ir cells--probably they remain under the greater influence of a system other than the cholinergic system. PMID- 11905255 TI - The role of IRBs in research involving commercial biobanks. PMID- 11905256 TI - Assisted reproduction: children conceived posthumously entitled to inheritance rights. PMID- 11905257 TI - Research guidelines: Hopkins deals with research volunteer death and aftermath. PMID- 11905258 TI - Privacy: intrahospital communications regarding patient care held constitutional. PMID- 11905259 TI - Constitutional law: use of force by mental health workers violated due process. PMID- 11905260 TI - Malpractice: suit against hospital for vicarious liability allowed despite settlement with physician. PMID- 11905261 TI - Taxation: tax-exempt status denied to HMOs owned by tax-exempt corporation. PMID- 11905262 TI - FDA: approval of generic drug reversed by U.S. Court of Appeals. PMID- 11905263 TI - Antitrust: medical device corporation liable for violating FTC order. PMID- 11905264 TI - The "disparate impact" argument reconsidered: making room for justice in the assisted suicide debate. PMID- 11905265 TI - Legislating privilege. PMID- 11905266 TI - The need to specify the difference "difference" makes. PMID- 11905267 TI - Pediatric research regulations under legal scrutiny: Grimes narrows their interpretation. PMID- 11905268 TI - In defense of the Hopkins Lead Abatement Studies. PMID- 11905269 TI - Placebo surgery for Parkinson's disease: do the benefits outweigh the risks? PMID- 11905270 TI - Should we impose quotas? Evaluating the "disparate impact" argument against legalization of assisted suicide. PMID- 11905272 TI - Untreated addiction imposes an ethical bar to recruiting addicts for non therapeutic studies of addictive drugs. PMID- 11905271 TI - I need a placebo like I need a hole in the head. PMID- 11905273 TI - Health-care rationing: critical features, ordinary language, and meaning. PMID- 11905274 TI - The physician as gatekeeper to the use of genetic information in the criminal justice system. PMID- 11905275 TI - Why The Netherlands? PMID- 11905276 TI - Capital planning: competing in the marketplace. AB - If you want to be successful, you need to spend money on improvements. As soon as you anticipate the need for a capital investment, start planning and exploring your financial opportunities. PMID- 11905277 TI - Back to basics 1. Employee retention and turnover: holding managers accountable. PMID- 11905278 TI - Rekindling the flame. Achieving community health through strong leadership. AB - Is the commitment to community health waning as hospitals focus their attention on dwindling reimbursement and proliferating regulations? Not if these four health care organizations are any indication of the innovative ways hospitals and communities can work together. PMID- 11905279 TI - Strategic planning. Design and construction: getting boards involved from the start. PMID- 11905280 TI - Wanted: a few good trustees. AB - As health care governance becomes increasingly complex and time-consuming, recruiting new trustees is more difficult. The solution? Open your mind to different options, including compensation, going outside the community, and making your meetings shorter and more efficient. PMID- 11905282 TI - Listening to patients: vital to nursing care. PMID- 11905281 TI - "Night of terror:" preventing patient self-harm. PMID- 11905283 TI - Hand infection associated with clenched fist syndrome in residents of long-term care facilities. AB - 1. Clenched fist syndrome is a clinical entity in which no organic disease can be found. 2. The syndrome usually follows a minor inciting incident and is associated with swelling, pain, and paradoxical stiffness. 3. Patients with severe forms of clenched fist syndrome may experience infections of the palm or joints. PMID- 11905284 TI - Pathological gambling. Identification and treatment. AB - 1. Social gamblers view gambling as a form of entertainment or recreation and gamble with no harmful effects, while problem gamblers' behavior causes disruption or harm to themselves or others in major life areas. Pathological gamblers fail to resist the impulse to gamble, with the resulting loss of control in their gambling behavior. 2. Pathological gambling is a primary mental health disorder of impulse control. 3. Treatment for pathological gamblers should be individualized, and interventions should address both the gambling disorder and any comorbid disorders. PMID- 11905286 TI - Art therapy. Enhancing psychosocial nursing. AB - 1. The creative process can be a means of reconciling conflicts and increasing awareness. 2. Art provides a permanent representation of clients' internal states and reflects the changes seen in the therapeutic progression. 3. Many psychological states lend themselves to a much greater extent to graphic representation, as opposed to verbalization. 4. Art holds the potential for enhancing the quality of therapy for clients by increasing expression and facilitating insight. PMID- 11905285 TI - Validation of the behavioral observation system. Using nurse ratings of acuity. AB - 1. One of the goals of the BOS is to gather supplemental, objective data to help unit administration determine patient acuity, and thus, staffing needs. 2. The BOS was developed to help provide information that is easily collected, objective, reliable, and valid. 3. The current study sought to establish preliminary support for use of the BOS to assess patient acuity and determine staffing needs. 4. Because the BOS can be completed as part of HSWs' daily tasks with little additional cost, the data can be available to decision makers to determine the unit's needs more accurately and efficiently. PMID- 11905287 TI - A pediatrician's view. Ticks! PMID- 11905288 TI - The emergence of tick-borne diseases. PMID- 11905289 TI - Rocky Mountain spotted fever: a review for the pediatrician. PMID- 11905290 TI - Lyme disease: fact versus fiction. PMID- 11905291 TI - Human monocytic ehrlichiosis: similar to Rocky Mountain spotted fever but different. PMID- 11905292 TI - Tularemia: lymphadenitis with a twist. PMID- 11905293 TI - Babesiosis: similar to malaria but different. PMID- 11905294 TI - New developments in tick-borne infections. PMID- 11905295 TI - Resident's column: spots and stripes. PMID- 11905296 TI - The use of "race" in research. PMID- 11905297 TI - Representations of race, ethnicity, and social class in case examples in The American Journal of Occupational Therapy. AB - OBJECTIVE: This article examines descriptors of race, ethnicity, and social class in case material in 145 articles published in The American Journal of Occupational Therapy from 1975 to 1998. METHOD: Ethnicity labels and descriptors of occupation or other indicators of social class in case examples describing adults were collected. Frequencies of these labels and descriptors were compared with the population demographics of the 1990 U.S. census using chi-square goodness-of-fit analyses. RESULTS: Reported indicators of race and social class were inconsistent and primarily absent in the literature, making comparisons with the U.S. population difficult. When missing race labels were assumed to be White, the case material showed disproportionate racial distributions compared with that of the U.S. population, with persons of minority races being significantly underrepresented. CONCLUSION: The findings suggest that appropriate representation of race and social class in occupational therapy literature has not yet been achieved. The authors suggest a change in documentation conventions for occupational therapy literature and clinical writing. PMID- 11905298 TI - Occupational therapy's dance with diversity. AB - As the demographics of the United States continue to change and we become a more pluralistic society, the increased diversity of the occupational therapy workforce and our consumers calls for an examination of the profession's stance on multiculturalism and diversity. Using the metaphor of dance, this article identifies the dance partners as the organization's leaders and its members. A historical review of the profession from the 1940s to the present traces the partners' steps to determine which led the dance of diversity during the profession's development. In this review, I discovered that the period when the profession most effectively and productively explored issues of diversity was during the early- to mid-1990s--a time when the organization and its members worked in harmony. At that time, occupational therapy's dance with diversity flowed with rhythm and synchronicity. PMID- 11905299 TI - Functional outcomes and daily life activities of African-American elders after hospitalization. AB - OBJECTIVE: African-American elders were recruited from a transitional unit after hospitalization and tracked for 6 months in the community after discharge to (a) examine functional outcomes on the unit and in the community and (b) identify patterns of participation in daily life activities. The International Classification of Impairments, Disabilities, and Handicaps (ICIDH-2) framework provided the structure to examine the connections among body systems, functional outcomes, and social participation for this population that has been underrepresented in past research. METHOD: A mixed design combined qualitative and quantitative methods, including qualitative interviews to document personal adaptive experience, a standardized functional assessment to identify functional outcomes, and a structured format to record activity participation. RESULTS: Findings revealed that 11 of the 17 participants improved their functional outcomes after discharge. Three patterns of activity participation identified were self-care, self-care and household management, and mixed activities. Contextual influences were diverse family support arrangements. CONCLUSION: Complex relationships were identified among body systems, functional outcomes, and daily life activities that were influenced by individual values and support arrangements. PMID- 11905300 TI - Ecological synergies in two groups of zoo chimpanzees: divergent patterns of time use. AB - This study investigated how two groups of zoo chimpanzees, one that lived in a small and mostly barren physical habitat yet had a successful social history and the other that lived in a larger and more enriched physical habitat yet had a turbulent social history, invested time occupationally. Different ecological synergies were found to emerge from these respective conditions and to influence time use in highly particularistic ways. As related to considerations of adaptedness, the limits of enriched physical environs when coupled with a dearth of social facilitation were revealed, as were the limits of social facilitation within impoverished physical spaces. Findings suggest that occupational therapists' expertise in activity analysis could be enlarged to encompass analysis of naturally socially embedded ways of doing things across multiple contexts. To advance the ethic of occupational justice, functional assessments of individuals could also be expanded into functional assessments of the occupational aliveness of proximate life environments. PMID- 11905301 TI - Young children's occupations: explicating the dynamics of developmental processes. AB - Describing development as sequential sensorimotor, cognitive, and psychosocial milestones does not explain how children learn to do or improve their occupations. In response to changes within occupational therapy and challenges in early intervention, this article focuses on development of occupation and proposes a dynamic system perspective. Contemporary research suggests that processes are in place to ensure that children develop as occupational beings. Social participation and caregiving routines guide the child toward becoming occupational by introducing intentional acts and by endowing experiences with meaning. A self-organizing process enables children to integrate their immature capacities and engage in occupations. Simultaneously, caregiving shapes these emerging behaviors, ensuring that performance is culturally compatible. Finally, using self-organization to maintain occupational engagement enables the child to accommodate to maturing abilities and environmental challenges. This reorganized pattern in turn is the basis for developmental changes, new behaviors, and refinement of current abilities. Application of concepts is made to acquisition of skills for eating a meal. PMID- 11905302 TI - Range of motion at the wrist: a comparison study of four wrist extension orthoses and the free hand. AB - OBJECTIVE: This study compared the total wrist range of motion permitted by four different styles of wrist extension orthoses and the free hand. METHOD: Using a repeated-measures, counterbalanced design, 40 healthy female volunteers 20 to 39 years of age shot a basketball while free handed and while wearing each of four wrist extension orthoses: AlignRite; Rolyan D-Ring Long; Rolyan D-Ring Short; and a custom-made, thumb hole design orthosis. The motion at the wrist was measured by an electrogoniometer. RESULTS: No significant differences were found in total wrist motion permitted among the four orthotic conditions. Analyses revealed that the custom-made orthosis allowed significantly less palmar flexion and significantly more dorsiflexion than the three commercially available orthoses. All orthoses significantly restricted wrist movement compared with the free hand. CONCLUSION: The commercially available wrist extension orthoses offered little difference in the amount of restriction they provided. The custom orthosis restricted movement to a different portion of the available range than did the commercial orthoses. Future research should examine how different strapping techniques on custom-made orthoses affect total range of motion permitted at the wrist. Knowledge of patterns of restriction among various styles of orthoses will help therapists to select the most appropriate orthosis for a client's individual needs. PMID- 11905303 TI - Measuring interface pressure: a laboratory-based investigation into the effects of repositioning and sitting. AB - Measurement of interface (or contact) pressure is important in assessing tissue viability in relation to pressure sore prevention and may be achieved through pressure mapping techniques. This article reports on two pilot studies using the Force Sensing Array pressure mapping system in a laboratory setting. The purpose of Study 1 was to examine the consistency of readings from the system across 1 min trials of repositioning, and Study 2 aimed to investigate changes in interface readings over a 20-min sitting period. Analyses on measurements of average pressure (mean of all sensor values) and maximum pressure (highest individual sensor value) were performed using the t test and repeated-measures analysis of variance. The results demonstrated that the use of average and maximum pressure measurements reflected only low reliability and that 6 min was likely to be the optimal sitting time required before stable pressure measurement. However, because of the limitations of using small convenience samples of healthy participants (n = 44 for Study 1, n = 20 for Study 2), these studies should be replicated with larger samples of healthy participants and then verified with disabled populations before adoption into clinical practice. PMID- 11905304 TI - Cognitive impairment after unilateral hemispheric injury of congenital or adult origin. AB - OBJECTIVE: The purpose of this study was to assess and compare cognitive functioning in adults with unilateral hemispheric injury due to either congenital damage or an ischemic event in young adulthood. METHOD: Adults with cerebral palsy resulting from left hemispheric brain damage were compared with adults who had a unilateral stroke in either the left or the right hemisphere. Our primary interest was to determine the impact on hemispheric dominance as revealed by dichotic listening, a task that assesses the bias for preferential listening and processing of sounds. Performance also was determined on a language-related task (word finding) and a spatial task (dot localization). RESULTS: Scores on the Quick Neurological Screening Test indicated that all participants demonstrated significant neuromotor deficits, whereas scores on the Barthel Index indicated that the participants were functional in basic activities of daily living. On cognitive assessments, healthy control participants demonstrated a pronounced left-hemisphere dominance and right-ear advantage; participants with injury to the left hemisphere showed a strong shift toward a right-hemisphere and left-ear dominance. In particular, injury of congenital origin appeared to foster this neural reorganization and localization of language-related functions into the healthy hemisphere. This shift was associated with a deterioration of performance on both the language and the spatial tasks. CONCLUSION: The importance of appreciating subtle deficits after unilateral injury is important in therapy. The dichotic listening test may provide a simple and useful means for evaluating persistent unilateral brain dysfunction in the clinical setting. PMID- 11905305 TI - The adolescent role assessment: psychometric properties and theoretical usefulness. AB - OBJECTIVE: Fostering career adaptability may improve vocational outcomes for adolescents. This study examined the responses of adolescents on the Adolescent Role Assessment (ARA) to explore its usefulness as a measure of career adaptability. METHOD: The ARA was administered to 101 adolescents 12 to 17 years of age. Descriptive statistics, reliability analysis, and factor analysis were generated from the ARA scores. Dominant narrative themes were coded into the six domains of the ARA and into the domains identified in the factor analysis. RESULTS: The internal consistency of the ARA was low with few differences in findings across age. The identified factors--developing aspirations, self efficacy, interpersonal competencies, and autonomy--are consistent with constructs of career adaptability found in the literature and differentiated low and high scorers. CONCLUSION: Major changes to the ARA content and scaling or development of a new assessment of career adaptability using the factor structure are recommended for clinical and research applications. PMID- 11905307 TI - Aspects of emic and etic measurement: lessons from Mary Poppins. AB - This article describes emic and etic approaches to measurement in terms of a process of associating indicators and constructs. In this process, it is important to establish evidence of an adequate range of indicators and a relevant association of indicators and constructs and to recognize that such evidence places bounds on the interpretation of measurement outcomes. A passage from one of P. L. Travers's Mary Poppins stories provides a counterpoint for this article's discussion of these key aspects of emic and etic methods of measuring human individuality. PMID- 11905306 TI - Classroom-based assessment: validation for the school AMPS. AB - The role of the occupational therapist working in the school system is to facilitate a student's task performance or ability to do purposeful and meaningful activities so that the student benefits from the educational experience. To fulfill this role, occupational therapists need assessments that address functional performance issues in the classroom and provide information for effective programming and consultation. The School Version of the Assessment of Motor and Process Skills (School AMPS) is an observational evaluation of functional skills in the classroom. Common classroom activities are observed to assess a child's school motor and school process skills. This study examined the validity of the School AMPS by comparing it to the Peabody Developmental Motor Scale-Fine Motor (PDMS-FM), a common assessment used in local area school districts in Edmonton, Alberta. Results show a higher correlation of the PDMS-FM with the motor scale of the School AMPS than with the process scale of the School AMPS, which was expected. The School AMPS appears to be a promising instrument for measuring the constructs of school motor and school process skills within a naturalistic setting. PMID- 11905308 TI - Learning from evidence: service outcomes and client satisfaction with occupational therapy home-based services. PMID- 11905309 TI - Using the Internet as a vehicle for research. PMID- 11905310 TI - Two studies suggest that relationship may be overlooked in practice and research. PMID- 11905311 TI - What's wrong with this picture? PMID- 11905312 TI - Lateral mandibulectomy and partial glossectomy with plate application. AB - The goals of composite resection with reconstruction plate application include removal of the primary tumor, any compromised portions of the mandible, and any involved lymph-bearing tissue. Recent advances in surgical technique and reconstruction have made this treatment a more appealing choice for patients. Although microvascular free flaps may be the treatment of choice in the younger patient with an excellent prognosis, the use of reconstruction plates with a myocutaneous flap remains a viable alternative for many patients with oral cancer. Regardless of the reconstructive technique utilized, both functional and aesthetic parameters must be addressed in treatment planning for patients with head and neck cancer. PMID- 11905313 TI - Modified radical and selective neck dissections. PMID- 11905314 TI - The pectoralis major myocutaneous flap for reconstruction of soft-tissue oncologic defects. PMID- 11905316 TI - Segmental resection of the anterior mandibular arch with fibular microvascular reconstruction. AB - Segmental mandibular resection should be undertaken if there is a demonstrable invasion by cancer or if such invasion is suspected. Before the advent of microvascular free flaps, immediate osseous reconstruction of the anterior mandible was technically demanding and unreliable. Mandibular reconstruction with microvascular free flaps is no longer a novelty, nor is it reserved for unusually large resections. It has become the method of choice for the immediate reconstruction of defects resulting from mandibular cancer. This technique is particularly well-suited for defects of the anterior mandibular arch. PMID- 11905315 TI - Anterior floor of mouth resection with marginal mandibulectomy. PMID- 11905317 TI - Maxillectomy. PMID- 11905318 TI - Radical neck dissection. PMID- 11905319 TI - Crown lengthening in the esthetic zone. AB - Crown lengthening in the esthetic zone is a prosthodontically designed and surgically executed procedure that must only be considered after careful restorative and surgical treatment planning, including a detailed smile analysis, clinical and radiographic evaluation of the quality of soft and hard tissues, and selection of the appropriate approach for each individual case. The presented techniques are modifications of the original conventional surgical approach, where longer healing periods may have been required and loss of papilla height or fullness is of concern. Recognition of the advantages and disadvantages of each technique should increase predictability and success in interdisciplinary smile enhancement therapy. PMID- 11905320 TI - De-epithelialized connective tissue pedicle graft: the palatal roll. PMID- 11905321 TI - A subepithelial connective tissue graft procedure for optimum root coverage. PMID- 11905322 TI - The semilunar flap technique for root coverage. AB - The semilunar flap technique may be considered as a simple, minimally invasive option for coronally repositioning the receded free gingival margin on the facial aspects of natural teeth and implants. Judicious case selection and careful application of the described procedure may enhance the final esthetic outcome using this approach. Work is currently in progress by the authors to provide long term evidence-based results using this technique, with future considerations including the application of recent advances in microsurgical instrumentation to facilitate surgical tissue management. PMID- 11905323 TI - Preserving alveolar ridge anatomy following tooth removal in conjunction with immediate implant placement. The Bio-Col technique. PMID- 11905324 TI - Preserving alveolar ridge anatomy following tooth removal in conjunction with delayed implant placement. AB - 1. Prior to placing implants, the health of the adjacent teeth and supporting tissues should be good. The presence of acute infection (exudate, granulation tissue, gingival hyperplasia), poor oral hygiene, or active chronic periodontal disease are associated with an increased incidence of infection or tissue breakdown. 2. Avoid excessive dissection of papilla. 3. Stabilize membranes with apical tacks and secure the coronal portion to the implant with the cover screw. 4. Relieve the periosteum to allow for a tension-free closure. 5. Avoid postoperative trauma on the surgical site. PMID- 11905325 TI - Use of the epithelialized palatal graft with dental implants. PMID- 11905326 TI - Subepithelial connective tissue grafting with dental implants. PMID- 11905327 TI - Treatment planning and vector analysis of mandibular distraction osteogenesis. PMID- 11905328 TI - Intraoral distraction osteogenesis: maxillary and mandibular lengthening. PMID- 11905329 TI - Maxillary distraction osteogenesis with rigid external distraction. PMID- 11905330 TI - Distraction osteogenesis for mandibular widening. PMID- 11905331 TI - Distraction osteogenesis for dental implants. PMID- 11905332 TI - Transport distraction: mandibular reconstruction. PMID- 11905333 TI - Distraction osteogenesis for congenital mandibular deformities. PMID- 11905334 TI - Treatment planning decisions: an overview. PMID- 11905335 TI - Surgical and orthodontic considerations for distraction osteogenesis with ROD appliances. AB - Tooth-borne/hybrid distraction devices offer a new approach for intraoral DO and better control of the distracted bony segments. These appliances also offer a unique characteristic by using the teeth as reference points for obtaining parallelism of the distraction devices to each other and to the vector of distraction in bilateral cases. More studies may be needed for long-term evaluation of the obtained results with intraoral distraction devices as well as biomechanical analysis of stress distribution throughout the mandible after distraction. PMID- 11905336 TI - Realities of craniofacial growth modification. AB - Facial growth modification can be an effective method of resolving skeletal discrepancies. There still is much controversy regarding our understanding of the nature and extent of skeletal orthopedic change possible in individual patients and the most effective appliances and timing of such treatment. In the treatment of class II patients, growth modification can lead to an improvement, if not complete correction of the class II malocclusion. Although two-phase treatment with an early first prepubertal phase can be effective, a later single-phase approach during early puberty seems to be equally effective. Certainly, before surgical correction of the mild to moderate skeletal class II problem in a growing patient is considered, an orthopedic phase of treatment prior to the pubertal growth spurt is an appropriate first step. Skeletal class III patients with a maxillary deficiency stand to gain significant benefits from early orthopedic treatment. However, such therapy may produce more favorable changes for older children and adolescents than previously thought. Nevertheless, orthopedic correction of the mild to moderate skeletal class III should be accompanied by regular progress evaluations to avoid creating significant dental compensations in the face with little skeletal change that ultimately requires surgery anyway. Skeletal class III patients with mandibular excess and/or vertical excess are poor candidates for growth modification. Orthopedic palatal expansion appears to be effective and stable at any time prior to late puberty, a stage of development when ossification of the maxillary sutures is more advanced. Consequently, the timing for expansion may be better determined by the specific needs of each patient. A functional shift resulting from a crossbite is optimally corrected early, so that asymmetric growth of the mandible can be reduced or even prevented. Postpubertal orthopedic expansion is likely to result in bone bending, which will reverse itself over time, potentially leaving periodontal compromise of the maxillary posterior teeth. Therefore, surgically assisted expansion should be considered in such cases. Vertical maxillary excess is challenging to treat with growth modification. The combination of interocclusal acrylic bite blocks combined with a high-pull headgear presently appears to be the best means available. Unfortunately, this treatment has limited success, it must be continued over many years owing to the long-term nature of vertical growth, and it is counterproductive in achieving a balanced sagittal jaw relationship in class I or class III patients with vertical excess. Future research will permit us to have a greater understanding of the nature and extent of facial growth modification possible in individual patients and the optimal appliances and timing of treatment to achieve the best outcome. The development of intraoral osseointegrated attachments such as implants and onplants hold promise for a future means of dissipating orthopedic forces to prevent unwanted dentoalveolar changes that presently occur with our tooth-borne appliances. Analogous attachments presently are undergoing clinical testing for surgically assisted orthopedic movements associated with distraction osteogenesis. PMID- 11905337 TI - Onplant-supported orthodontic anchorage. AB - The use of onplant-supported orthodontic anchorage is still in its early stages. The concepts presented in this article have tried to stress the importance of anchorage control in orthodontic cases, some problems associated with traditional means to augment orthodontic anchorage, the rationale for developing a skeletal anchorage unit based on endosseous implant technology, and the effectiveness of the onplant as an anchorage unit. Cephalometric superimpositions are most likely the best method to determine the effectiveness of the onplant's contribution to anchorage control and have been included here to that end. Further research into its effectiveness may broaden the use of onplants in future orthodontic cases. PMID- 11905338 TI - Presurgical orthodontics for orthognathic surgery. PMID- 11905339 TI - Orthodontic and surgical camouflage for treatment of skeletal discrepancies. AB - Masking procedures can be used for a variety of situations and must be tailored to the patient's individual needs. Although they can result in esthetic changes for the patient, they do little for functional concerns. The most commonly used procedure is a bony augmentation genioplasty. It is most effective when a patient is genially deficient and has an overjet in the range of 3 to 5 mm. PMID- 11905340 TI - Applied surgical anatomy for mandibular fracture management. PMID- 11905341 TI - Treatment of mandibular-condylar fractures. AB - Particularly with true dislocation fractures, nonoperative treatment with maxillomandibular fixation followed by physiotherapeutic exercises leads to poor results, as was proved with axiography and clinical examinations. The main reason for this is the shortening and scarring of the condyloid process and the lack of function of the lateral pterygoid muscle. The condyle with its insertion of the muscle is usually displaced medially and anterially and nearly in touch with the origin on the pterygoid process so that protrusion by the muscle is no longer possible. The physiologic relationship of the lateral pterygoid muscle is restored after reduction of the condyle and osteosynthesis of the condylar neck fracture and the original distance between origin and insertion of the muscle is re-established and is a fundamental necessity for regaining function (Fig. 40). The anchor screw osteosynthesis is a most effective technique with low limitations for its indication. A comparison with plates shows this technique to be very economic because one anchor screw has the effect of at least one five hole plate with five plating screws. That means a reduction of osteosynthesis implants of up to 80%, which saves a lot of money. On the other hand, the sophisticated technique of an anchor screw osteosynthesis needs some training on the part of the surgeon to get the best results possible. In general, we could realize that the anchor screw osteosynthesis gives a perfect adaptation of the fracture ends with compression also on the inner cortical layer, which with plates is only possible in rare cases. After an osteosynthesis of mandibular condyle neck fractures with an axial anchor-screw there are a few cases with an absorptive process in the fracture interface where the screw migrates in an axial direction with loosening of the osteosynthesis. This effect can be compared with the effect of a dynamic hip screw, which leads to compression of the callus, which speeds up bony union at the expense of shortening the bone. When the same absorption happens using a plate, the fracture ends cannot become sintered and the plate is in danger of fracturing as a result of metal fatigue. Ceipek evaluated 136 patients with mandibular condylar neck fractures treated with axial anchor screw osteosynthesis. Thirty-six of these screws showed signs of migration, but only 3.7% for more than 4 mm. For the migration process there are some important risk factors: difficult repositioning of the proximal fragment, dorsal luxation fracture, indirect method of anchor screw osteosynthesis, narrow condyle neck, no intercuspation in the molar region, no compliance, and disturbance of bone healing. Another stable technique of osteosynthesis should be used if patients show more risk than one risk factor. PMID- 11905342 TI - Treatment of comminuted fractures of the mandible. AB - Management of comminuted mandible fractures requires careful planning. Treatment is based on a thorough understanding of the medical management of severely injured patients, biology of these injuries, the biomechanics of the mandible, and principles of fracture fixation. PMID- 11905343 TI - Management of complications associated with mandible fracture treatment. PMID- 11905344 TI - Symphysis and parasymphysis fractures. PMID- 11905345 TI - Management of mandibular body fractures. PMID- 11905346 TI - Treatment of mandibular angle fractures. PMID- 11905347 TI - Surgical management of parotid disease. PMID- 11905348 TI - The current status and possible future for lithotripsy of salivary calculi. AB - Extracorporeal and intracorporeal shockwave lithotripsy are effective in at least 30% of cases, particularly when combined with adjuvant techniques. Factors that appear to affect outcome adversely are stone size, partial fragmentation with reduced clearance, duct stenosis, and poor gland function. PMID- 11905349 TI - The surgical management of submandibular gland disease. PMID- 11905350 TI - The surgical management of salivary gland disease of the sublingual gland and floor of mouth. PMID- 11905351 TI - The management of minor salivary gland tumors of the oral cavity. PMID- 11905352 TI - Surgery of the salivary ducts. AB - Successful surgery of the salivary ducts relies on an understanding of the surrounding anatomy and the delicate dissection of tissues in order to reduce morbidity. Trauma to the ducts should be assessed when lacerations or wounds encroach on their paths. Early diagnosis and treatment will reduce the complications of stricture and fistula formation from these injuries. Sialoliths can be located in several places along the length of the salivary ducts. The correct diagnosis and positioning of the stone in the duct is important in establishing the appropriate surgical approach. Imaging using plain films, ultrasonography, and endoscopy can be very valuable, with sialography and CT scans helpful in cases of radiolucent stones, glandular atrophy, or suspected tumor. As the condition becomes more chronic, resulting in glandular atrophy, excision of the diseased gland is often indicated. Treatment of excessive salivary flow in patients with cerebral palsy can be managed by a combination of ductal repositioning and glandular excision. Redirection of both the parotid and submandibular glands can be accomplished, either to reroute excess salivary flow or salvage the duct in cases of lesion excision. PMID- 11905353 TI - Isolated nasal fractures. PMID- 11905354 TI - Orbital fractures. PMID- 11905355 TI - Treatment of nasal-orbital-ethmoid fractures. PMID- 11905357 TI - Frontal sinus reconstruction. PMID- 11905356 TI - Associated soft tissue injuries. PMID- 11905358 TI - Anatomic considerations in the partially and fully edentulous mandible. PMID- 11905359 TI - Calvarial bone harvest and grafting techniques for maxillary and mandibular implant surgery. PMID- 11905360 TI - Vascularized free bone grafts for maxillary and mandibular reconstruction. AB - Vascularized osteocutaneous free grafts have been successfully applied to maxillofacial reconstruction. Careful planning allows for implant prosthesis fabrication in these cases. The scapula, fibula, and ilium are the donor sites that result in the most favorable application of implant prosthetics. Intraoral soft tissues, although compromised, are manageable with meticulous home oral hygiene practices and frequent recall examinations with professional cleaning. Many patients severely impaired by trauma, neoplasm, or congenital deformity can have restoration of form and function with the amalgamation of microsurgical and osteointegration techniques. PMID- 11905361 TI - Skeletal-dental reconstruction of the compromised mandible with composite bone grafts. PMID- 11905362 TI - Anatomic considerations in the partially and fully edentulous maxilla. PMID- 11905364 TI - Maxillary sinus bone grafting. AB - When a patient is missing posterior teeth and desires a fixed prosthesis, bone grafting is often needed. Since 1983, our clinical experience indicates that sinus grafting can provide very good restoration of the posterior quadrants. If the implants are loaded in a physiologically stable manner, one may expect maintenance of bone levels. PMID- 11905363 TI - Skeletal-dental reconstruction of the compromised maxilla with composite bone grafts. PMID- 11905365 TI - Placement of implants into extraction sites. PMID- 11905366 TI - Surgical approaches to repositioning of the inferior alveolar nerve for placement of osseointegrated implants. AB - Severe resorption of the posterior mandible poses one of the most difficult restorative challenges to the implant surgeon today. Although bone augmentation procedures may increase the amount of bone in which to place implants, these procedures are costly and have an increased risk of complications to the patient. With the proper evaluation, treatment planning, and attention to technical detail, nerve repositioning appears to offer an excellent alternative to augmentation procedures for placement of dental implants. PMID- 11905367 TI - Guided tissue regeneration. PMID- 11905369 TI - Browlift. PMID- 11905368 TI - Chemical skin resurfacing. PMID- 11905370 TI - Nasal surgery for the aging face. PMID- 11905372 TI - Hair restoration surgery. Hair transplantation and micrografting. PMID- 11905371 TI - CO2 laser skin resurfacing for the aging face. PMID- 11905373 TI - Scar modification. Techniques for revision and camouflage. AB - The surgery and management of scars is a protracted and staged process that includes preparation of the skin through hygienic measures, scar softening (if indicated) with steroids, massage and pressure dressings, skilled execution of the surgical plan, and thorough postoperative wound care. This process generally covers a 1-year period for the various stages mentioned. Many general host and local skin factors will directly affect the final revision result. The two most important indirect factors that the surgeon must endeavor to control are optimal patient preparation and cutaneous health, and patient compliance with, and an ability to carry out, those wound care measures that the surgeon prescribes. Keloid and burn contracture scars represent two entities that are complicated and challenging to treat owing to their abnormal morphophysiologic features. Management of these scars is prolonged, and the patient must understand that the ultimate result will usually be a compromise. New grafting techniques, such as cultured autodermal grafts, offer improved initial management of burn wounds that may subsequently optimize scar revision in these patients. Keloids, and to a lesser extent hypertrophic scars, require steroid injections, pressure treatment, careful surgery, and protracted wound support and pressure treatment (exceeding 6 months) after surgery. PMID- 11905374 TI - Liposculpture of the cervicofacial region. PMID- 11905375 TI - Blepharoplasty: an overview. PMID- 11905376 TI - Best days. PMID- 11905377 TI - Whatever it takes. PMID- 11905378 TI - Destigmatizing mental illness. PMID- 11905379 TI - Mobilizing, together, for mental health. Interviewed by Kate O'Reily. PMID- 11905380 TI - A vision for mental health care. Interviewed by Lisa Legge. PMID- 11905381 TI - One to three: eliminating the stigma of mental illness. PMID- 11905382 TI - Study slams Joint Commission's approach to measuring quality. PMID- 11905383 TI - Remember diplomacy in peer review process. PMID- 11905384 TI - CQI 'business as usual'? Not for patient safety. PMID- 11905385 TI - Press, Ganey winners are best in customer care. PMID- 11905386 TI - Are you supporting the silent incident victims? PMID- 11905387 TI - Bioterrorism watch. Anthrax aftermath: adverse drug reactions, vaccine controversy undercut CDC extended treatment offer. PMID- 11905388 TI - Toward a national health risk management approach in Australia. AB - There has been increasing international consensus about the importance of competition for achieving national growth and community well-being. The Australian government accordingly has introduced policies to promote such competition. Major legislative review and many public inquiries have assisted implementation of national competition policy and the development of national goals and standards related to international agreements to promote health and sustainable development. Since the 1980s, Australia has had legislation that requires the identification and control of health risks arising at work. The management structures necessary for coordinated delivery of national programs designed for effective identification and control of health risks arising in communities to achieve national health and development goals are still being developed, however. Major difficulties related to this development are discussed. National health development programs should be approached primarily through establishment of regional partnerships between bodies responsible for managing community health, local government, and employment placement, in consultation with other relevant organizations and the community. Related research and evaluation programs are required. PMID- 11905389 TI - Providing evidence of good allied health care for the veteran population: development of a unique management system. AB - A unique management system was developed to enhance the quality of allied health care for the elderly. The management system is a multipronged framework of processes and outcomes, which address key stakeholder needs, and is based on best scientific evidence. Use of the management system provides efficiencies for funding bodies in data collection and monitoring service quality. It also confirms the professional integrity of the allied health service provider and underscores the importance of client-therapist partnership in determining appropriate health outcomes. Further testing is required to assess the relationship between use of the allied health management system and better quality service provision, cost containment, and consistently good client health outcomes. PMID- 11905391 TI - Allied health professional services for oncology outpatients: an Australian comparative study. AB - Psychosocial interventions used by the allied health team are now seen as making an important contribution to improving the psychological and functional statues of the increasing numbers of patients coping with cancer. However, there is a call from within the Allied Health Professionals (AHP) literature for empirical research to ensure that service delivery is supported by a reliable understanding of the needs of consumers. The present discussion responds to such a call by presenting the comparative findings from two independent studies looking at the needs of oncology out-patients for AHP services in Australia. The first study was completed at the Oncology Day Care Ward at the Mater Hospital, Brisbane, Queensland. The Brisbane research was subsequently repeated at the Mater Hospital, Newcastle, New South Wales. It is the comparative findings from the two projects that are presented here. The impetus for both studies came from a keen desire by the AHP teams of both hospitals to better understand quality of life issues in relation to their oncology outpatients in order to be able to provide appropriate and effective services. The same research instrument was used for both studies so the data are directly comparable. It is the hope and expectation that these findings will be useful to inform the work of other AHP teams who are engaging in the challenging but rewarding task of understanding and meeting the needs of oncology patients and their families for holistic care. PMID- 11905390 TI - Professional perceptions about home safety: cross-national validation of the Home Falls and Accidents Screening Tool (HOME FAST). AB - The Home Falls and Accidents Screening Tool (HOME FAST) was developed to measure the risk of older people falling within their home environment. If this tool is to be effective, the underlying dimensions perceived by potential raters when using the HOME FAST need to be consistent with the purpose of the tool. The content validation process undertaken to evaluate the HOME FAST and a method to develop a home safety score are described. Experts in home safety assessment were recruited from the British Association of Occupational Therapy, the Chartered Society of Physiotherapists, and the Royal College of Nursing in the United Kingdom. Participants rated each HOME FAST item using a Thurstone technique format. Each item was weighted according to the level of perceived falls risk each expert attributed to the home safety item. Principal components factor analysis identified a two-factor structure interpreted as environment and function underlying the HOME FAST. Results indicated that no item should be deleted from the checklist. Weights were calculated for each item to generate an overall hazard score. Some differences in responses to the degree of risk associated with home safety items were noted between the professional groups. The HOME FAST has captured highly relevant home safety items considered by an expert panel and measures a domain applicable to home safety and falls risk. PMID- 11905392 TI - Occupational therapy practice in acute care neurology and orthopaedics. AB - The purpose of this research was to develop a consensus description of occupational therapy practice in acute care orthopaedics and neurology by Australian therapists. The Delphi technique was used to develop consensus concerning therapists' aims for their patients, the interventions they used, and the factors affecting their practice in acute care. Therapists' primary aim was to conduct a thorough assessment for referral and further treatment purposes. Therapists in acute hospital neurology focus on developing maximal independence in self-care activities and preventing deformities, followed by preparations for discharge or transfer to rehabilitation. Therapists with an orthopaedic caseload state independence in self-care as their second most important aim, but the ranking of intervention methods indicates preparation for discharge in various ways takes precedence over direct self-care retraining. This difference may indicate a disparity between practice ideals and the realities of the acute care setting. The most important factor affecting practice in acute care is early referral for occupational therapy services. Recommendations are made for managers and educators to ensure the most effective practice in acute care are made. PMID- 11905393 TI - Heteroglossia: voices heard on Capitol Hill from a Bakhtinian perspective. PMID- 11905394 TI - Physical therapists' knowledge, advice, and administration of nonprescription medications to their clients. AB - Physical therapists often have limited knowledge and little formal training in pharmacology, yet they frequently advise their clients on the use of over-the counter (OTC) medications and administer these in the course of treatment. This practice may have a bearing on professional liability if side effects and contraindications are not adequately considered. In addition, some nonprescription medications may have an effect on physical therapy care. The purpose of this study was to assess physical therapists' knowledge about OTC medications, and the extent to which they advise clients about OTC medications and administer them. To examine these issues, a self-administered, anonymous questionnaire was mailed to a random sample of 25% of clinical physical therapists registered in New South Wales, Australia (n = 660). It was found that a substantial proportion of practicing physical therapists advised and administered OTC medications despite their limited training and knowledge in the area. PMID- 11905395 TI - [Interactions of risk factors in chronic diseases]. PMID- 11905396 TI - [Determination of influenza vaccination in persons 65 years of age and older]. AB - BACKGROUND: Influenza vaccination is an effective preventive intervention to reduce the negative consequences of this disease. The objective of this work is to describe population patterns of use of influenza vaccination, to analyze the effect of a set of variables on the likelihood of being vaccinated and of having a proactive attitude towards the vaccine in the age group of 65 year and over. METHODS: Cross sectional study. From the Spanish National Health Survey of 1997 those 65 and more year old were selected. The association of reporting having had an influenza vaccination last as year or having asked for the vaccine was estimated. RESULTS: A total sample of 1,148 was analyzed. Overall, 51.3% of subjects reported having received a vaccination last year. 18.0% of them reported having asked for the vaccine. Odds Ratios statistically associated with the vaccination were: 70 to 74 year 1.6 (1.2-2.2), 75 and older 2.0 (1.5-2.8), male 1.4 (1.1-1.9), residents in towns with less than 10,001 inhabitants 1.6 (1.0 2.6), non smokers 2.1 (1.3-3.5), with a last visit to a physician in the last two weeks 1.8 (1.3-2.6). Odds Ratios statistically associated with having a proactive attitude towards the vaccine were: finished studies with more than 15 year 1.8 (1.1-2.9), no consumption of medicine in the last two weeks 1.9 (1.1-3.6), and self-reported health as good or very good 1.7 (1.1-2.9). CONCLUSIONS: Overall influenza vaccination levels are not appropriate. Several factors have been identified as barriers for receiving this effective intervention. No association was found with socio-economic status. Some differences were also found regarding having a positive attitude towards the vaccine. This study may contribute to identifying population groups who are not receiving influenza vaccination and for designing strategies aimed to enhance influenza vaccination among them. PMID- 11905397 TI - [Mortality attributable to tobacco consumption in the years 1987 and 1997 in Castilla la Mancha, Spain]. AB - BACKGROUND: Smoking is an important public health problem and is one of the main avoidable causes of morbidity and early mortality. The aim of this work was to describe the mortality attributable to tobacco consumption in Castilla la Mancha between 1987 and 1997. METHOD: Deaths in relation to age, sex and cause of death were obtained from the Death's Register of Castilla la Mancha. From the National Health Surveys of 1987 and 1997, the percentages of non-smokers, smokers and ex smokers in the population in relation to age and sex were recorded. The relative risks of death were obtained from the Cancer Prevention Study II, carried out in the United States. The proportion of deaths attributable to smoking was calculated for each year, and according to sex and age group, from the etiological fraction of the population. Likewise, loss of potential life in years and the mean number of years of potential life lost were also calculated. RESULTS: During the study period, 18% of all the deaths in Castilla la Mancha can be attributed to tobacco consumption. Mortality is higher in males than in females, and the most important diagnostic categories were tracheobronchopulmonary cancer (24.3%) in males and diseases of the cardiovascular system (24.28%) in females. These were also the conditions most responsible for years of potential life lost. CONCLUSIONS: Every day, in Castilla la Mancha, 8 people die from smoking-related conditions. The measures currently in practise to control tobacco consumption are insufficient. PMID- 11905398 TI - [Results of the first program to reduce the prevalence of smoking in the staff of Salamanca Council, Spain]. AB - BACKGROUND: Smoking is the most important risk factor for public health and one of the factors with the greatest economic repercussions for society. The aim of the present work is to publish the results of a programme to reduce smoking in the working environment. METHODS: A programme was designed in two stages. The first consisted of carrying out a survey to establish the prevalence of smoking and the attitudes towards smoking of staff working for Salamanca district Council. The second stage consisted of a specialized treatment programme for all the workers who wanted to try to give up smoking. RESULTS: A total of 384 workers answered the questionnaire. Of these, 135 (35.1%) were smokers of whom 80.5% (n = 113) wanted to give up smoking and to receive medical help to do so. A total of 73 workers started the programme. After one year of treatment, 41% of the workers who had started the programme had dropped out. An average of 2.9 of every 3 workers found the programme satisfactory. CONCLUSIONS: The prevalence of smoking in Salamanca District Council is similar to that described in the National Health Survey, 1997. Programmes to treat smoking in the working environment are useful to reduce the prevalence of smoking and are welcomed by the workers. Reducing the prevalence of smoking is the first step towards achieving smoke-free institutions. PMID- 11905399 TI - [Non-typhoid salmonellosis in in a basic health area of Navarra, Spain]. AB - BACKGROUND: Salmonella gastro-enteritis is a Zoonoses transmitted by the ingestion of food products and water or fomites contaminated by the faeces of infected people or animals. At present, constitutes a world-wide pandemic. The aim of the present study has been to in progress examine cases of non-typhoidal salmonellosis in the Health Area I of Navarra (376,079 inhabitants). METHODS: 39,697 outpatient specimens submitted for culture during 1993-2000 were analysed retrospectively. Standard procedures to isolate enteropathogens were employed. The Salmonella strains were serotyped. Data was collected on age, sex, specimen date and result of culture and antimicrobial susceptibility testing for all isolates. RESULTS: 2,924 salmonellae were isolated (7.4%) with the most frequent serotype being Salmonella Enteritidis (62%). The highest isolation rate was associated with children, particularly infants (1,117.3 per 100,000 inhabitants). Salmonella Typhimurium was typically more resistant than Salmonella Enteritidis, although resistance rates in both have increased in recent time. CONCLUSIONS: In spite of the socio-economic improvements, the incidence of gastro-enteritis associated with Salmonella spp. has continued to increase in recent years, mainly affecting infants, and constitutes an important public health problem. PMID- 11905400 TI - [Accidents attended in a basic health area of Girona, Spain]. AB - BACKGROUND: Accidents have been largely unstudied in the area of Primary Care. They are one of the most frequent motives for consultation in the Emergency Services and the first assistance that accident victims receive is usually in primary care centres. Establishment of the incidence and clinicoepidemiological characteristics of the accidents attended in a Basic Health Area can provide important information about which of these could be susceptible to preventive actions. METHODS: DESIGN: descriptive study. LOCATION: primary care: SAMPLE: all the patients attended for accidents (389) in the Primary Care Centre between October 1998 and May 1999. VARIABLES: age, sex, place of the accident, type of lesion, location of lesions, agents involved, intentionality, complementary tests, treatment and referral. STATISTICAL ANALYSIS: estimation of means, standard deviation, proportions and 95% confidence intervals. RESULTS: Incidence: 4.1% (CI95%: 3.7-4.5%). Sex: males 59% (CI95%: 54.2-64%) and females 40.9% (CI95%: 36-45.8%). Age: younger than 20 years, 50.4% (CI95%: 45.4-55.4%). Most common activity associated with accidents: leisure 24.4% (CI95%: 20.2-28.7%). PLACE: home 36.2% (C95%: 31.5-41%). Most frequent lesion: contusion 39.6% (CI95%: 34.7-44.4%). Most frequent site of lesion: arms 37.5% (CI95%: 32.7-42.3%). Most common agent involved: tools and machinery 15.9% (CI95%: 12.3-19.6%). Of these, 92.2% (CI95%: 89.3-94.7%) were accidental. Type of visit: 83.3% (CI95%: 79.6-87%) were attended as emergencies; 79.5% (CI95%: 75.4-83.5%) received treatment with dressings and/or medication. Of these, 9.8% (CI95%: 6.8-12.7%) required referral to a hospital, 13.3% (CI95%: 10-16.7%) required complementary tests. CONCLUSIONS: Most accidents occur in young people and educational campaigns to prevent accidents and directed towards this population group are clearly needed. PMID- 11905401 TI - [Association between clustering of cardiovascular risk factors and the risk of cardiovascular disease]. AB - BACKGROUND: Cardiovascular diseases are the main cause of mortality in Spain. The aim of this work was to study the association between clustering of cardiovascular risk factors and the risk of suffering major cardiovascular events: ischemic cardiopathy, cerebrovascular disease and peripheral arteriopathy of the lower limbs. METHOD: A descriptive transversal study was carried out in a city health centre, with a total of 2248 patients selected by simple random sampling of the clinical records with a mean age of 15 years. The data were obtained by examining the clinical records and estimating Odds Ratios (OR) for any cardiovascular event (n = 224), ischemic cardiopathy (n = 123), cerebrovascular disease (n = 84) and peripheral arteriopathy (n = 55) in relation to the number of cardiovascular risk factors. The cardiovascular risk factors included in the study were smoking, arterial hypertension, hypercholesterolemia, hypertriglyceridemia, diabetes and obesity. The OR was adjusted for age and sex. RESULTS: The percentage of patients with 0, 1, 2, 3 and 4-6 cardiovascular risk factors was 39.1, 32.8, 17.5, 6.9 and 3.7 respectively. The OR for experiencing a cardiovascular event associated to 1, 2, 3 and 4-6 cardiovascular risk factors was 1.6 (CI95%: 0.9-2.7), 2.8 (CI95%: 1.7-4.7), 3.6 (CI95%: 1.9-6.5) and 5.6 (CI95%: 2.9-10.8), respectively. The OR for ischemic cardiopathy associated to the same risk levels were 2.3 (CI95%: 1.1-4.6), 2.5 (CI95%: 1.2-5.2), 5.3 (CI95%: 2.4-11.5) and 6.2 (CI95%: 2.7-14.3), respectively. For cardiovascular disease, the OR were 1.1 (CI95%: 0.5-2.5), 2.3 (CI95%: 1.2-5.3), 2.4 (CI95%: 1.0-5.9) and 5.6 (CI95%: 2.2-14.1), respectively. The OR for peripheral arteriopathy were 2.1 (CI95%: 0.8-5.9), 3.7 (CI95%: 1.3-10.5), 3.3 (CI95%: 1.0-11.1) and 6.1 (CI95%: 1.8-20.3), respectively. CONCLUSIONS: The addition of cardiovascular risk factors is associated with an increased risk of cardiovascular events. This finding emphasises the need for prevention of cardiovascular risk factors in primary care. PMID- 11905402 TI - Effect of nitric oxide on the expression of insulin-like growth factors and the insulin-like growth factor binding proteins throughout the lifespan of the human corpus luteum. AB - The presence of insulin-like growth factors (IGF), IGF binding proteins (IGFBP) and IGF receptor type 1 (IGF-IR) in the human corpus luteum was investigated by examining the expression and production of related proteins throughout the lifespan of the corpus luteum and the action of nitric oxide upon their production. The expression of proteins in corpora lutea from the early, mid-and late luteal phases was assessed by immunohisto-chemistry, evaluated by a semi quantitative analysis and the functional study was performed in corpus luteum explants incubated with nitric oxide donors. IGF-I and -II and IGFBP-1 and -3 were measured in the culture media by specific immunoassays. The results showed that IGF-I and -II, IGFBP-1 to -6 and IGF-IR were detected in the human corpus luteum throughout the luteal phase. Moreover, the expression and production of IGF-I and IGFBP-1 increased progressively from corpora lutea from the early to late luteal phases (P < 0.05), whereas the expression and production of IGFBP-2, 4 and -5 were significantly higher in corpora lutea from the mid-luteal phase (P < 0.05). No differences were observed in the expression of IGF-II, IGFBP-3 and -6 and IGF-IR throughout the lifespan of the corpus luteum. However, functional studies showed that nitric oxide donors elicited a stimulatory action on production of IGF-I in corpora lutea from the early luteal phase (80%) and on production of IGFBP-1 in corpora lutea from the late luteal phase (50%) (P < 0.05), whereas production of IGF-II and IGFBP-3 was not affected by nitric oxide. In conclusion, the components of the IGF-IGFBP system are expressed in the human corpus luteum throughout its lifespan. Nitric oxide regulates IGF-I and IGFBP-1 production, indicating that the growth factors may serve, at least in part, as mediators of the action of nitric oxide in the human corpus luteum. PMID- 11905403 TI - Expression of uterine and cervical epithelial cadherin during relaxin-induced growth in pigs. AB - Epithelial cadherin (E-cadherin), a member of the cadherin family of calcium dependent adhesion molecules, is present in reproductive tissues. Relaxin, a hormone important for uterine and cervical growth in pigs, increases the expression of E-cadherin in the MCF-7 mammary epithelial cell line. The objective of this study was to characterize the expression of E-cadherin during relaxin induced growth of the uterus and cervix in an immature pig model, independent of high circulating steroids. After administration of relaxin to prepubertal gilts, the uterus and cervix were collected. E-cadherin mRNA and protein were measured by northern and western blot analysis, respectively. A 120 kDa protein band, corresponding to E-cadherin, was detected in all tissues examined. Relaxin significantly (P < 0.05) increased the amount of E-cadherin protein in the uterus (P < 0.05), whereas no significant changes were observed in E-cadherin protein in the cervix. A 4.2 kb E-cadherin transcript was detected in all tissues and E cadherin mRNA was significantly higher (P < 0.05) in uteri from relaxin-treated gilts compared with control gilts. E-cadherin was localized by immunocytochemistry to the epithelial cells of the uterine and cervical lumen, and the uterine glandular epithelium. Quantitative analysis revealed that administration of relaxin significantly increased (P < 0.05) the height of the uterine luminal epithelium compared with that of the controls. This is the first report of the expression of E-cadherin in the uterus and cervix of pigs. The findings from this study indicate that relaxin increases the expression of uterine E-cadherin in the reproductive tract of pigs. Administration of relaxin to prepubertal gilts in vivo increased uterine epithelial cell growth independent of circulating steroids, with a concomitant increase in E-cadherin expression. PMID- 11905404 TI - Adrenomedullin, calcitonin gene-related peptide and their receptors: evidence for a decreased placental mRNA content in preeclampsia and HELLP syndrome. AB - OBJECTIVE: The human placenta expresses a variety of vasoactive substances and neuropeptides, which play an important role in the regulation of placental blood flow in both the maternal and foetal compartment and are therefore of critical importance for foetal growth and development. Our study was planned to examine placental mRNA amounts of vasodilatory adrenomedullin (AM), calcitonin gene related peptide (CGRP) and their receptors (AM-R and CGRP-R) in preeclampsia and HELLP syndrome (hemolysis, elevated liver enzymes, low platelets). These are severe maternal conditions leading to an altered uteroplacental and fetoplacental perfusion and a higher risk for foetal growth retardation, premature delivery, infant mortality, and even maternal death. STUDY DESIGN: We included 17 patients with preeclampsia, four women with HELLP syndrome and 34 controls. After delivery, the mRNA levels of AM, AM-R, CGRP, CGRP-R, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and -actin were measured in placental villi and chorionic plates using quantitative real-time PCR. RESULTS: AM/-actin and AM/GAPDH mRNA ratios were significantly lower in placental villi in preeclampsia than in controls (P<0.05) as were CGRP/-actin and CGRP/GAPDH mRNA ratios in chorionic plates (P<0.05). Placental AM-R and CGRP-R mRNA amounts were unaffected. CONCLUSION: Our data show a reduction of AM and CGRP mRNAs in contrast to unchanged mRNA levels of their receptors in placenta specimens of women with preeclampsia or HELLP syndrome. PMID- 11905405 TI - Oxygen therapy for hypercapnic patients with chronic obstructive pulmonary disease and acute respiratory failure: a randomized, controlled pilot study. AB - OBJECTIVE: To investigate the effect of oxygen therapy on outcome and on symptomatic hypercapnia. DESIGN: Randomized, controlled, single-blind study. SETTING: Multidisciplinary intensive care unit of a university teaching hospital. PATIENTS: Patients admitted with a clinical diagnosis of an acute exacerbation of chronic obstructive pulmonary disease and a PaO2 <6.6 kPa (50 mm Hg) and PaCO2 >6.6 kPa (50 mm Hg) on air. INTERVENTIONS: Patients received oxygen therapy titrated to increase arterial oxygen tension to >6.6 kPa (50 mm Hg) or >9 kPa (70 mm Hg). Patients in the low-oxygen tension group also received doxapram if they developed an acidosis with pH <7.2, whereas those in the high-oxygen tension group received doxapram if they developed symptomatic acidosis. Bronchodilator, steroid, and antibiotic therapy was standardized. MEASUREMENTS AND MAIN RESULTS: Two patients in the low-oxygen tension group (n = 17) required mechanical ventilation and another one died. No patients in the high-oxygen group (n = 17) had a poor outcome, but this difference was not significant. No patient in either group became comatose or developed an acute cardiac arrhythmia. CONCLUSIONS: Traditional teaching related to oxygen therapy for hypercapnic patients with an acute exacerbation of chronic obstructive pulmonary disease may be incorrect. A large randomized, controlled study is required to confirm this impression. PMID- 11905406 TI - Easy access to pediatric intensive care unit care: an urban legend? PMID- 11905407 TI - No effect of preoperative selective gut decontamination on endotoxemia and cytokine activation during cardiopulmonary bypass: a randomized, placebo controlled study. AB - BACKGROUND: Cardiopulmonary bypass predisposes the splanchnic region to inadequate perfusion and increases in gut permeability. Related to these changes, circulating endotoxin has been shown to rise during cardiac surgery, and may contribute to cytokine activation, high oxygen consumption, and fever ("postperfusion syndrome"). To a large extent, free endotoxin in the gut is a product of the proliferation of aerobic gram-negative bacteria and may be reduced by nonabsorbable antibiotics. OBJECTIVE: To evaluate the effect of preoperative selective gut decontamination (SGD) on the incidence of endotoxemia and cytokine activation in patients undergoing open heart surgery. DESIGN: Prospective, randomized, placebo-controlled double-blind trial. SETTING: Tertiary-care university teaching hospital. INTERVENTION: Preoperative administration for 5 to 7 days of oral nonabsorbable antibiotics (polymyxin B and neomycin) vs. placebo. The efficacy of SGD was assessed by culture of rectal swabs. PATIENTS: Forty-four patients (median age 65 yrs, 29 males) were included in a pilot study to establish the sampling points of perioperative measurements. Seventy-eight consecutive patients (median age 65 yrs, 55 males) were enrolled for the prospective study; of these, 51 were randomly allocated to take SGD (n = 24) or placebo (n = 27); 27 were included in a control group (no medication). MEASUREMENTS AND RESULTS: SGD but not placebo effectively reduced the number of rectal swabs that grew aerobic gram-negative bacteria (27% vs. 93%, respectively; p < .001). SGD did not affect the occurrence of perioperative endotoxemia, nor did it reduce the tumor necrosis factor-alpha, interleukin-10, or interleukin-6 concentrations (p > .20), as determined before surgery, upon aorta declamping, 30 mins into reperfusion, or 2 hrs after surgery. Also, SGD did not alter the incidence of postoperative fever or clinical outcome measures such as duration of artificial ventilation and intensive care unit and hospital stay. CONCLUSION: SGD effectively reduces the aerobic gram-negative bowel flora in cardiac surgery patients but fails to affect the incidence of perioperative endotoxemia and cytokine activation during cardiopulmonary bypass and the occurrence of a postperfusion syndrome. PMID- 11905408 TI - Characterization of an N-terminal secreted domain of the type-1 human metabotropic glutamate receptor produced by a mammalian cell line. AB - A Chinese hamster ovary cell line has been established which secretes the N terminal domain of human mGlu1 receptor. The secreted protein has been modified to contain a C-terminal hexa-histidine tag and can be purified by metal-chelate chromatography to yield a protein with an apparent molecular weight of 130 kDa. Following treatment with dithiothreitol the apparent molecular weight is reduced to 75 kDa showing that the protein is a disulphide-bonded dimer. N-terminal protein sequencing of both the reduced and unreduced forms of the protein yielded identical sequences, confirming that they were derived from the same protein, and identifying the site of signal-peptide cleavage of the receptor as residue 32 in the predicted amino acid sequence. Endoglycosidase treatment of the secreted and intracellular forms of the protein showed that the latter was present as an endoglycosidase H-sensitive dimer, indicating that dimerization is taking place in the endoplasmic reticulum. Characterization of the binding of [3H]quisqualic acid showed that the protein was secreted at levels of up to 2.4 pmol/mL and the secreted protein has a Kd of 5.6 +/- 1.8 nm compared with 10 +/- 1 nm for baby hamster kidney (BHK)-mGlu1alpha receptor-expressing cell membranes. The secreted protein maintained a pharmacological profile similar to that of the native receptor and the binding of glutamate and quisqualate were unaffected by changes in Ca2+ concentration. PMID- 11905409 TI - ACOG practice bulletin. Perinatal viral and parasitic infections. Number 20, September 2000. (Replaces educational bulletin number 177, February 1993). American College of Obstetrics and Gynecologists. PMID- 11905410 TI - Diffuse malignant epithelial mesotheliomas of the peritoneum in women: a clinicopathologic study of 25 patients. AB - BACKGROUND: The behavior of diffuse peritoneal mesotheliomas in women and the possible relation between tumor morphology and outcome are uncertain. Reported survival has ranged from < 1 month to > 14 years, and a previous study found that tumor morphology could not be used reliably for predicting outcome. The authors examined the behavior of diffuse epithelial peritoneal mesotheliomas in women and the possible relation between pathologic features and outcome. METHODS: Twenty five female patients with diffuse peritoneal epithelial malignant mesotheliomas were divided into two groups: those who survived for < 4 years (60%) and those who survived for > 4 years (40%). Both groups were compared in terms of age, presentation, treatment, survival, tumor architecture, mitotic rate, necrosis, nuclear grade, and immunohistochemical profile. RESULTS: Patients in the two groups were similar in terms of age at diagnosis (median ages, 50.7 years and 49.9 years), presentation, initial tumor burden, and treatment. In both groups, the most common initial clinical presenting features were ascites and abdominal pain. The tumors typically took the form of multiple nodules measuring < 1.5 cm in greatest dimension. Slightly less than 50% of patients in both groups received some form of chemotherapy or radiation therapy after undergoing tumor-reductive surgery or biopsy. Overall survival ranged from 1 month to 15 years. The median survival was 12 months in the group of women who survived for < 4 years and 7 years in the group of women who survived for > 4 years. Overall, 10 of 25 patients survived for > or = 5 years. One patient was alive with disease 15 years after diagnosis. Although there was a suggestion that the tumors in patients with short survival more often had solid architecture and high-grade nuclei, these findings were not significant statistically. The frequency of necrosis and the mitotic activity were the same in both groups. CONCLUSIONS: The spectrum of diffuse epithelial peritoneal mesotheliomas in women includes tumors that are highly aggressive and behave much like pleural mesotheliomas, although a sizeable number of tumors, unlike the pleural tumors, are relatively indolent. However, because there do not appear to be morphologic features that reliably identify favorable tumors versus unfavorable tumors, aggressive therapy for all women with diffuse peritoneal mesotheliomas may be warranted. PMID- 11905411 TI - Central nervous system metastasis from nasopharyngeal carcinoma: a report of two patients and a review of the literature. AB - BACKGROUND: Central nervous system (CNS) metastasis from nasopharyngeal carcinoma (NPC) is an extremely rare occurrence, although direct intracranial invasion is not infrequent in patients with NPC at a locally advanced stage. Only five other patients have been reported in detail in the English literature. METHODS: The clinical records of two such patients with NPC who were diagnosed with metastasis to the spinal cord (intradural) and to the occipital lobe, respectively, were reviewed. The literature was searched for a review of similar incidents. RESULTS: Both patients had locally advanced disease at the time of presentation and were treated with neoadjuvant chemotherapy and radical radiotherapy. The CNS metastases in both patients were accompanied by disease recurrences in multiple sites after a prolonged period of clinical remission. Spread through cerebral spinal fluid was postulated for the patient with spinal cord metastasis, and hematogenous spread was postulated for the patient with brain metastasis. Aggressive surgical resection with or without postoperative radiotherapy conferred reasonable survival and symptom control. The patient with brain metastasis died 6 months later of lung metastasis, whereas the other patient is still alive 40 months from the diagnosis of spinal metastasis. CONCLUSIONS: Good symptom control and disease control can be achieved for patients with CNS metastasis after surgery with or without radiotherapy. After aggressive therapy, the ultimate survival depends on control of extracranial disease. PMID- 11905412 TI - Factors predicting response and survival in 149 patients with unresectable hepatocellular carcinoma treated by combination cisplatin, interferon-alpha, doxorubicin and 5-fluorouracil chemotherapy. AB - BACKGROUND: The objective of the current study was to identify patient and disease related factors that influence response and survival for patients with unresectable hepatocellular carcinoma (HCC) who received a systemic combination chemotherapy consisting of cisplatin, alpha-interferon, doxorubicin, and 5 fluorouracil (PIAF). METHODS: From July 1996 to February 1999, 149 patients with unresectable HCC were treated with PIAF: cisplatin (20mg/m2 intravenously, Days 1 4), doxorubicin (40mg/m2 intravenously, Day 1), 5-fluorouracil (400mg/m2 intravenously, Days 1-4), and alpha-interferon (5MU/m2 subcutaneously, Days 1-4), once every 3 weeks up to a maximum of six cycles. Univariate and multivariate analyses of patient and disease characteristics were used to identify factors predicting response and survival. RESULTS: The objective response rate according to conventional criteria was 16.8% (complete response in 3 out of 149 patients, or 2%, 95% confidence interval [CI] 0-4.3%; partial response in 22 out of 149 patients, or 14.8%, 95% CI 9-20%). The median survival time was 30.9 weeks (95% CI 22.1 to 40). Significant independent predictors of an objective response were: absence of cirrhosis (P = 0.006), low bilirubin level (P = 0.006), and positive hepatitis C serology (P = 0.025). The following factors were related to a shorter survival time: high Okuda stage (P = 0.001), vascular involvement (P = 0.018), and cirrhosis (P = 0.008). Good risk patients (absence of cirrhosis and total bilirubin < or = 0.6mg/dL) had an objective response rate of 50%. CONCLUSIONS. Patients with unresectable HCC who also have normal total bilirubin and non cirrhotic livers have a better chance of response and prolonged survival after treatment with systemic PIAF. PMID- 11905413 TI - Primary pleuropulmonary synovial sarcoma: reappraisal of a recently described anatomic subset. AB - BACKGROUND: Primary pleuropulmonary synovial sarcoma (SS) is a rare neoplasm and a recently recognized anatomic subset. Its clinicopathologic attributes are not yet well defined. METHODS: In this study, the clinical and histopathologic features of 12 SS arising in the lung and/or pleura were analyzed. RESULTS: The neoplasms occurred in 7 men and 5 women, 20-72 years old (median, 31 years), were well circumscribed with a mean size of 7.2 cm, and involved either lung (9 cases), pleura (2 cases), or both (1 case). All the tumors were of monophasic type. Nine showed a classic spindle cell pattern, and three showed predominantly poorly differentiated features. All but one case showed at least focal positivity for epithelial membrane antigen (EMA), a finding characteristic of this tumor. The lack of EMA staining in one case, proven by electron microscopy to be SS, was attributed to the scarcity of material available for immunohistochemical stains. The diagnosis was proven cytogenetically in three cases. Within 2 years, local recurrence developed in 8 patients (75%), 3 of whom developed metastasis (25%). Five patients died of their disease within 2.5 years, 4 of them from uncontrolled local disease. CONCLUSIONS: The authors conclude that pleuropulmonary SS, although rare, represents a distinct anatomic subset having pathologic features similar to those of its soft tissue counterpart. Its clinical behavior appears more aggressive, perhaps because of relatively later presentation combined with the difficulty in obtaining a wide surgical margin. PMID- 11905414 TI - Vulvar melanoma: is there a role for sentinel lymph node biopsy? AB - BACKGROUND: The objective of this study was to evaluate the author's recent, preliminary experience with the sentinel lymph node procedure in patients with vulvar melanoma and to compare this experience with treatment and follow-up of patients with vulvar melanomas who were treated previously at their institution. METHODS: From 1997, sentinel lymph node procedure with the combined technique (99mTechnetium-labeled nanocolloid and Patente Blue-V) was performed as a standard staging procedure for patients with vulvar melanoma with a thickness > 1 mm and no clinically suspicious inguinofemoral lymph nodes. For the current study, clinicopathologic data from all 33 patients with vulvar melanoma who were treated between 1978 and 2000 at the University Hospital Groningen were reviewed and analyzed. RESULTS: From January 1997 until December 2000, identification of sentinel lymph nodes was successful in all nine patients who were referred for treatment of vulvar melanoma. Three patients underwent subsequent complete inguinofemoral lymphadenectomy because of metastatic sentinel lymph nodes. In follow-up, groin recurrences (in-transit metastases) occurred in two of nine patients, both 12 months after primary treatment. Both patients had melanomas with a thickness > 4 mm and previously had negative sentinel lymph nodes. There was a trend toward more frequent groin recurrences in patients after undergoing the sentinel lymph node procedure (2 of 9 patients) compared with 24 historic control patients (0 of 24 patients; P = 0.06). Five of 33 patients developed local recurrences: Two patients had groin recurrences, and 11 patients developed distant metastases. Twelve patients died of vulvar melanoma. Seventeen patients with a median follow-up of 66 months (range, 9-123 months) are currently alive (overall survival rate, 52%). CONCLUSIONS: Although the numbers were small, this study showed that the sentinel lymph node procedure is capable of identifying patients who have occult lymph node metastases and who may benefit from lymphadenectomy for locoregional control and prevention of distant metastases. However, the data also suggest that the sentinel lymph node procedure may increase the risk of locoregional recurrences (in-transit metastases), especially in patients with thick melanomas. The potential role of the sentinel lymph node procedure as an alternative method of lymph node staging in patients with vulvar melanoma needs further investigation only within the protection of clinical trials and probably should be restricted to patients with melanomas with intermediate thickness (1-4 mm). PMID- 11905415 TI - It pays to be prepared: when you least expect it, someone may ask you about cancer genetics! PMID- 11905416 TI - Clinical pathways for managing patients receiving interleukin 2. AB - As biologic therapies enter the mainstream for cancer and HIV treatments, clinicians need the knowledge and expertise to safely and competently care for their patients who are undergoing these therapies. This article provides an overview of the immune system, emphasizing the elements that are affected by the biologic agent interleukin 2 (lL-2), lL-2 has been approved for use in the treatment of metastatic renal cell carcinoma and metastatic melanoma. Clinical trials currently are being conducted to determine its use in treating other cancers. The severity of side effects of lL-2 varies with the dose, route, and schedule of administration. The most common effects with all methods of administration are flu-like symptoms. Because the side effects of lL-2 are relatively predictable, clinical pathways offer practical tools for anticipating and managing the toxicities associated with lL-2 administration. PMID- 11905417 TI - [What made the elderly lady feel dizzy?]. PMID- 11905418 TI - Healthy solutions. PMID- 11905419 TI - Early diagnosis and prevention of colorectal cancer. The Third European Endoscopy Forum: conclusions of a symposium held in Faro, Portugal, June 8-10, 2001, with the help of an educational grant from Olympus. PMID- 11905421 TI - Tumour location and the effects of preoperative radiotherapy in the treatment of rectal cancer. PMID- 11905422 TI - Doctors and the law--a personal view. PMID- 11905423 TI - Doctors and the law--a personal view. PMID- 11905424 TI - Portsmouth POSSUM models for abdominal aortic aneurysm surgery (Portsmouth Physiological and Operative Severity Score for enUmeration of Morbidity and mortality). PMID- 11905425 TI - Randomized study comparing cardiac response in endovascular and open abdominal aortic aneurysm repair. PMID- 11905426 TI - Late effects of the treatment of childhood cancer on the female reproductive system and the potential for fertility preservation. PMID- 11905427 TI - Sperm motility in the semen analysis affects the outcome of superovulation intrauterine insemination in the treatment of infertile Asian couples with male factor infertility. AB - OBJECTIVE: To ascertain the relationship between the initial and unprocessed sperm parameters and pregnancy rates in SOIUI, for Asian couples with male factor infertility. DESIGN: Retrospective study. SETTING: A large government tertiary care women's hospital with 15,000 deliveries per year. POPULATION: One thousand four hundred and seventy nine couples undergoing 2846 cycles of SOIUI. METHODS: All couples enrolled in the SOIUI programme were analysed, comparing initial sperm parameters and the post-processed total motile sperm, against pregnancy rates per cycle. MAIN OUTCOME MEASURES: Pregnancy rates in relation to initial sperm parameters and post-processed total motile sperm. RESULTS: Ninety-three percent of the couples had male factor infertility. The average normal forms for these men was 14.7%. Overall pregnancy rate was 12.1% per completed SOIUI cycle. We found a significant drop in pregnancy rates if the percentage of motile sperms in the unprocessed sperm sample fell below 30%. We also found that insemination of at least 1 million motile sperm resulted in a significant increase in pregnancy rates. CONCLUSIONS: We recommend SOIUI as an effective treatment of suitable couples with male infertility, before embarking on IVF. However, if the initial percentage of motile sperm fell below 30%, or if after processing, the total motile sperm count was fewer than 1 million, these couples should consider in vitro fertilisation. PMID- 11905428 TI - Uterine artery embolisation for the treatment of symptomatic fibroids in 114 women: reduction in size of the fibroids and women's views of the success of the treatment. AB - OBJECTIVE: To assess the reduction in size of fibroids following uterine artery embolisation and to analyse women's views of the success of treatment. DESIGN: An uncontrolled case series of 114 consecutive women who underwent uterine artery embolisation for the treatment of fibroids over two years. SETTING: The Diagnostic and Interventional Radiology Department at The Royal Surrey County Hospital, Guildford, UK. METHODS: Bilateral uterine artery embolisation was performed for the treatment of symptomatic fibroids. Magnetic resonance imaging was carried out before and six months following embolisation. Women completed outcome questionnaires following their treatment. MAIN OUTCOME MEASURES: The sites. imaging signal characteristics and percentage reduction in the volume of three dominant fibroids were determined from the magnetic resonance scans. Outcome was measured by questionnaire. Women were asked whether their symptoms resolved completely, improved, remained unchanged or deteriorated. RESULTS: One hundred and sixty-five fibroids of 114 women (mean age 42) were analysed. Forty five percent of women had complex fibroid masses and 50% had fibroids > or =8.5cm in diameter. The median reduction in the fibroid volume was 58%. The median reduction of the volume of complex fibroid masses, submucous fibroids, fibroids > or =8.5cm and fibroids with high and low signal on T2 weighted sequences were 58%, 63%, 50%, 62% and 51%, respectively. Ninety-one percent of the women's symptoms had resolved or improved following embolisation. DISCUSSION: The majority of women were satisfied with their outcome. We have shown that uterine artery embolisation is a successful treatment for symptomatic fibroids of all types, sizes and signal characteristics. PMID- 11905429 TI - A randomised controlled trial of intramuscular syntometrine and intravenous oxytocin in the management of the third stage of labour. AB - OBJECTIVE: To compare the efficacy and safety of intravenous oxytocin with intramuscular syntometrine in the management of the third stage of labour. DESIGN: A prospective randomised trial. SETTING: A university teaching hospital. METHODS: A total of 991 women having a singleton pregnancy and vaginal delivery were randomised by a computer-generated number to receive either 1 ml syntometrine intramuscularly or 10 units of intravenous Syntocinon after delivery of the anterior shoulder of the fetus. MAIN OUTCOME MEASURES: Blood loss during delivery, rate of postpartum haemorrhage, need for repeated oxytocics, haemoglobin level before and 24 hours after delivery, duration of third stage, need for manual removal of placenta and sides effects including hypertension, nausea, vomiting, headache and chest pain. RESULTS: The use of intravenous oxytocin was associated with a reduction in postpartum blood loss (P < 0.001) but there was no difference in the risk of postpartum haemorrhage in the need for repeated oxytocic injections and the drop in peripartum haemoglobin level between the two groups. There was also no difference in the risk of prolonged third stage, or in the need for manual removal of placenta. The use of syntometrine was associated with a higher risk of hypertension (RR 2.39, 95% CI 1.00-5.70). Other side effects were mild in nature with no differences between the two groups. CONCLUSIONS: There are no important clinical differences in the effectiveness of intramuscular syntometrine and intravenous oxytocin for the prevention of postpartum blood loss. Intravenous oxytocin is less likely to cause hypertension. PMID- 11905430 TI - Prediction of chorionicity in twin pregnancies at 10-14 weeks of gestation. AB - OBJECTIVE: To examine the accuracy of sonographic determination of chorionicity in twin pregnancies at 10-14 weeks of gestation. DESIGN: Prospective study on the sonographic prediction of chorionicity at 10-14 weeks of gestation. PARTICIPANTS: During a 30 month period, from October 1997 to May 2000, 165 women attending the departments of fetal medicine or ultrasound. METHODS: Sonographic criteria used in the diagnosis of chorionicity were the number of placental sites, the lambda (lambda) and T signs and the thickness of the inter-twin membrane. The diagnosis of chorionicity was made at the time of the ultrasound examination using all these features and subsequently compared with the postnatal diagnosis, confirmed either by placental histology or discordancy in infant sex. RESULTS: In 150 cases with confirmation of chorionicity following delivery, 116 were postnatally classified as dichorionic and 34 monochorionic. Prenatal ultrasound examination correctly identified chorionicity in 149 (99.3%) cases. The most reliable indicator for dichorionicity was a combination using the lambda sign or two separate placentae with a sensitivity and specificity of 97.4% and 100%, respectively. The most useful test in predicting monochorionicity was the T sign with a sensitivity of 100% and specificity of 98.2%. Measurement of the inter twin membrane thickness was a less reliable indicator where the sensitivity for dichorionicity and specificity for monochorionicity was only 92.6%. CONCLUSIONS: Ultrasound examination of twin pregnancies at 10-14 weeks of gestation predicts chorionicity with a high degree of accuracy using a combination of the number of placentae, lambda and T signs and inter-twin membrane thickness. All hospitals should encourage departments providing ultrasound services to undertake chorionicity determination when examining women with twin pregnancies at this gestation. PMID- 11905431 TI - Acute tracheitis with severe upper airway obstruction complicating pregnancy. PMID- 11905432 TI - Clinical assessment of Er,Cr:YSGG laser application for cavity preparation. AB - In this study, an erbium,chromium:YSGG (Er,Cr:YSGG) laser emitting at a wavelength of 2.78 microm was clinically applied to remove caries and prepare cavities, and the clinical outcome was evaluated. Effective clinical application of Er,Cr:YSGG laser had been expected from previous studies. This study included 44 patients (26 females, 18 males; aged 23-58) with a total of 50 cavity preparations by the Er,Cr:YSGG laser irradiation at 3-6 W with water spray. Patient acceptance and prognosis were evaluated. Most cases (94%) were prepared without anesthesia, and no pain was felt in 34 cases (68%). No adverse reaction was observed in any of the cases, and patient acceptance for this system was favorable. All cases had a good prognosis. In 45 cases (90%), overall clinical evaluation was satisfactory. From the present study, it can be concluded that the Er,Cr:YSGG laser system is an efficient, effective, and safe device for caries removal and cavity preparation. PMID- 11905433 TI - Differential residual effects of zaleplon and zopiclone on actual driving: a comparison with a low dose of alcohol. AB - STUDY OBJECTIVES: To compare residual effects of zaleplon 10 mg, zopiclone 7.5 mg, and placebo, and a social dose of alcohol on car driving, memory, and psychomotor performance. DESIGN: Two-part placebo controlled, crossover study. Part 1 was single blind, Part 2 double blind. SETTING: University research institute. PARTICIPANTS: Thirty healthy volunteers (15 men and 15 women, mean age 32 +/- 7 years) INTERVENTIONS: In Part 1 alcohol and alcohol-placebo drinks were administered around noon. In Part 2 single oral doses of zaleplon 10 mg, zopiclone 7.5 mg and placebo were administered at bedtime. MEASUREMENT AND RESULTS: A highway driving test, laboratory tests of word learning, critical tracking and divided attention, and subjective assessments of sleep, mood, and effects of treatments on driving. Driving started 40 minutes after a second alcohol dose in Part 1, and 10 hours after drug intake in Part 2. The results demonstrated that alcohol, at average plasma concentrations of approximately 0.030 g/dl, significantly impaired performance in all tests. Zaleplon's residual effects did not differ significantly from those of placebo in any test. In contrast, zopiclone had significant residual effects on driving, divided attention, and memory. The magnitude of impairment in the driving test observed the morning after zopiclone 7.5 mg was twice that observed with alcohol. CONCLUSION: Zaleplon 10 mg has no residual effects on driving when taken at bedtime, 10 hours before driving. In contrast, zopiclone 7.5 mg can cause marked residual impairment. Patients should be advised to avoid driving the morning after zopiclone administration. PMID- 11905434 TI - Hysteroscopic insemination of mares with low numbers of nonsorted or flow sorted spermatozoa. AB - The objectives of this study were 1) to compare pregnancy rates resulting from 2 methods of insemination using low sperm numbers and 2) to compare pregnancy rates resulting from hysteroscopic insemination of 5 x 106 nonsorted and 5 x 106 spermatozoa sorted for X- and Y-chromosome-bearing populations (flow sorted). Semen was collected with an artificial vagina from 2 stallions of known acceptable fertility. Oestrus was synchronised (June to July) in 40 mares, age 3 10 years, by administering 10 ml altrenogest orally for 10 consecutive days, followed by 250 microg cloprostenol i.m. on Day 11. All mares were given 3000 iu hCG i.v. at the time of insemination to induce ovulation. Mares were assigned randomly to 1 of 3 treatment groups: mares in Treatment 1 (n = 10) were inseminated with 5 x 10(6) spermatozoa deposited deep into the uterine horn with the aid of ultrasonography. Mares in Treatment 2 (n = 10) were inseminated with 5 x 10(6) spermatozoa deposited onto the uterotubal junction papilla via hysteroscopic insemination. Mares in Treatment 3 (n = 20) were inseminated using the hysteroscopic technique with 5 x 10(6) flow sorted spermatozoa. Spermatozoa were stained with Hoechst 33342 and sorted into X- and Y-chromosome-bearing populations based on DNA content using an SX MoFlo sperm sorter. Pregnancy was determined ultrasonographically at 16 days postovulation. Hysteroscopic insemination resulted in more pregnancies (5/10 = 50%) than did the ultrasound guided technique (0/10 = 0%; P<0.05) when nonsorted sperm were inseminated. Pregnancy rates were not significantly lower (P>0.05) when hysteroscopic insemination was used for sorted (5/20 = 25%) and nonsorted spermatozoa (5/10 = 50%). Therefore, hysteroscopic insemination of low numbers of flow sorted stallion spermatozoa resulted in reasonable pregnancy rates. PMID- 11905435 TI - Biochemical development of subchondral bone from birth until age eleven months and the influence of physical activity. AB - Subchondral bone provides structural support to the overlying articular cartilage, and plays an important role in osteochondral diseases. There is growing insight that the mechanical features of bone are related to the biochemistry of the collagen network and the mineral content. In the present study, part of the normal developmental process and the influence of physical activity on biochemical composition of subchondral bone was studied. Water content, calcium content and characteristics of the collagen network (collagen, hydroxylysine, lysylpyridinoline (LP) and hydroxylysylpyridinoline (HP) crosslinking) of subchondral bone were measured in newborn foals, 5-month-old foals (pasture-grown and box-confined) and 11-month-old foals at 2 differently loaded sites of the proximal articular surface of the first phalanx. During the first 5 months postpartum, water and hydroxylysine content decreased significantly while calcium and collagen content and the amount of HP and LP crosslinks increased significantly. The withholding of physical activity during this developmental phase affected the biochemical characteristics of subchondral bone only at the site that is loaded during physical exercise. At this site, calcium content and both HP and LP crosslink levels increased significantly less than in pasture-raised animals. During development from 5-11 months, measured parameters remained essentially constant, except for water content, which decreased further. It is concluded that substantial changes, presumed to be largely exercise-driven, take place during the normal process of development in the biochemical composition of equine subchondral bone. Normal development of subchondral bone is presumably important for the normal functional adaptation of this bone to the loading conditions it is subjected to and therefore essential to resist the future biomechanical challenges the horse will encounter during its athletic career. The findings from this study and the assumed important role of subchondral bone quality in the pathogenesis of osteochondral disease merit more attention to the role of the collagen network in subchondral bone. PMID- 11905436 TI - Polysaccharide storage myopathy in the M. longissimus lumborum of showjumpers and dressage horses with back pain. AB - This study was designed to investigate whether horses with clinical signs of back pain due to suspected soft tissue injuries were affected by polysaccharide storage myopathy (PSSM). Diagnosis of PSSM in muscle biopsies obtained from the M. longissimus lumborum of 5 showjumpers and 4 dressage horses with a history of back pain is reported. M. longissimus lumborum biopsies of these horses were characterised histopathologically and in 3/9 cases also by electron microscopy. Observations were compared with M. gluteus biopsies of the same horses, and with M. gluteus biopsies obtained from 6 Standardbreds with recurrent exertional rhabdomyolysis and from 6 healthy trotters. M. longissimus biopsies from horses with back pain showed pathognomonic signs of PSSM, i.e. high glycogen and/or abnormal complex amylase-resistant polysaccharide deposits. Similar features were found in M. gluteus biopsies of the same horses. Sections of horses with rhabdomyolysis had increased PAS stain when compared with healthy horses, but did not show amylase-resistant material. Qualitative observations were corroborated by quantitative histochemistry (optical densities) of sections stained with PAS and amylase PAS. This study demonstrated the presence of PSSM in the M. longissimus of showjumpers and dressage horses with back pain and indicates that epaxial muscle biopsy is an option in diagnosing back problems in horses when clinical examination and imaging techniques do not provide a precise diagnosis. PMID- 11905437 TI - Effectiveness of a two-dose regimen of prostaglandin administration in inducing luteolysis without adverse side effects in mares. AB - Our objectives were to determine whether repeated administration of prostaglandin F2alpha (PGF2alpha) to simulate the endogenous mode of secretion would be more effective than a single injection in inducing luteolysis and enable use of smaller doses less likely to cause adverse side effects. The main study comprised 43 dioestrous mares, who were given im. either a single 10 mg dose of natural PGF2alpha (n = 22) or 2 doses of 0.5 mg PGF2, 24 h apart (n = 21). The intensity of side effects was assessed in 8 dioestrous mares given 5, 1.5, 0.5 or 0 mg PGF2alpha in consecutive cycles. Two doses of 0.5 mg PGF2alpha 24 h apart caused lysis of the corpus luteum in all mares, whether this was determined from a fall in plasma progesterone concentrations or reproductive tract/behavioural changes; and when 10 mg PGF2, was given, the corpus luteum was lysed in 17 of 22 mares i.e. a lower proportion (P = 0.0485). A single dose of 0.5 mg PGF2a was no more effective than saline in inducing luteolysis.The intensity of side effects of PGF2alpha increased with dose. Although the 0.5 mg dose was no more likely than saline to cause sweating or muscle spasms, it raised plasma cortisol concentrations and prevented the decline in heart rate seen after saline. We conclude that a 2 dose regimen of administration increases the luteolytic efficacy of PGF2alpha and thereby provides a way to minimise adverse side effects. PMID- 11905438 TI - Septic flexor tendon core lesions in five horses. PMID- 11905439 TI - Severe crushed chest injury with large flail segment: computed tomographic three dimensional reconstruction. PMID- 11905440 TI - Combined intraperitoneal and extraperitoneal rupture of bladder. PMID- 11905441 TI - Implementation of a monitoring system to measure impact of stormwater runoff infiltration. AB - Stormwater infiltration is a drainage mode, which is more and more used in urban areas in France. Given the characteristics of urban surfaces, and especially the loads of various pollutants contained in stormwater, it is important to assess the impact of stormwater infiltration systems on soil and groundwater by carrying out field experiments. The main difficulty is due to the complexity of the system observed and the need of multidisciplinary approaches. Another difficulty is that measurements are carried out in situ, in an uncontrolled environment submitted to quantitatively and qualitatively highly variable interferences. Very long term monitoring is needed to get representative results. In order to contribute to solve these problems, the OTHU project has recently been launched in Lyon (France). One of its key action concerns a long-term (10 years) experiment on an infiltration basin specifically rehabilitated for measurements and operational drainage issues. This paper presents the experimental site, the objectives of the project and the way the monitoring process has been built according to the various disciplines involved (biology, ecology, hydrology, chemistry and soil sciences) and to the will of assessing all the uncertainties in the measurement process. PMID- 11905442 TI - Performance of partially separate sewer systems and impacts on receiving waters. AB - The aim of this document is to present and discuss the results of the experimental work undertaken in Laje stream, in a section near the village of Oeiras in Portugal. The work was developed with the main objective of characterising stormwater quality in Portuguese drainage systems, and to predict the effects of the performance of partially separate sewer systems on receiving waters. For this purpose, volume and characteristics of stormwater carried by a partially separated sewer system were estimated, both in terms of flow and Chemical Oxygen Demand (COD) loads. The study also analyses the eventual implementation of non-conventional solutions, as a way of reducing problems of bacteriological contamination of seawaters. This aspect is particularly important in Portugal, where the population is mainly concentrated in urban areas located down-stream of important drainage basins, close to the coastline. Therefore, sanitary sewer overflows discharging directly into receiving waters are frequent, with possible consequences in terms of bacteriological contamination of bathing areas. Based on experimental research and available data it was possible to collect informations regarding stormwater average COD and overflow coliform loads, and the occurrence of first flush effects. PMID- 11905443 TI - Increasing wastewater system performance--the importance of interactions between sewerage and wastewater treatment. AB - The necessity to assess sewer systems and wastewater treatment plants (WWTPs) as integral parts of the wastewater system has been well known for several years and discussed in many conferences. Until recently, sewer systems and WWTPs were improved (or optimised) separately or independently, which resulted in suboptimal solutions. Nowadays, in The Netherlands as well as in other European countries, a trend can be recognised towards more integral solutions. Nevertheless, due to a lack of knowledge on the interactions between the sewer systems and the WWTPs the implementation of this way of thinking in practice takes a long time. This paper describes the results of two cases in which the interactions between sewerage and wastewater treatment are incorporated within the optimisation of a wastewater system. The first case illustrates the importance of taking the interactions into account, while the second case shows how to deal with the interactions within a wastewater system optimisation study. It is concluded that the combination of total wastewater system analysis, incorporating the interactions within the wastewater system, with efficient search algorithms is expected to be very valuable in future wastewater system optimisation studies. PMID- 11905444 TI - Poly-beta-hydroxybutyrate metabolism in dynamically fed mixed microbial cultures. AB - The kinetics of production and degradation of poly-beta-hydroxybutyrate (PHB) by a mixed activated sludge culture growing on acetate was studied in a sequencing batch reactor (SBR). Occasionally a very high amount of acetate was added to the steady state system in order to obtain high PHB concentrations in the cells (fPHB). This made it possible to follow PHB production and degradation over a wide range of fPHB-data (between 0 and 0.8 Cmol/Cmol). The results were compared with data available in literature and with equations derived by metabolic modeling. This led to some remarkable observations. For the feast period, the ratio q(feast)PHB/-q(feast)Ac (specific PHB production rate over specific acetate uptake rate) was used to indicate which fraction of the substrate is stored. Experimentally and theoretically it was shown that this ratio has a constant value for dynamically fed systems operated at a sludge retention time (SRT) > 2d. This value is 0.6 Cmol/Cmol under aerobic conditions and 0.4-0.5 Cmol/Cmol under anoxic conditions, irrespective of the specific growth rate of the biomass and the specific acetate uptake rate in the feast period. Degradation of internal stored PHB could be described with a first order degradation rate with respect to the PHB content of the cells. Degradation of PHB appeared to be independent of the type of electron acceptor present in the system and independent of the SRT of the system. The kinetic descriptions can be used to predict PHB production and consumption in general in dynamic fed wastewater treatment systems, and they provide some trends for modeling purposes. PMID- 11905445 TI - Denitrification of groundwater with elemental sulfur. AB - Autotrophic denitrification was studied in laboratory columns packed with granular elemental sulfur only and operated in an upflow mode. Soluble inorganic carbon, sodium bicarbonate, was supplied as source of carbon for microbial growth. Denitrification rates of up to 0.20 kg N removed m(-3) d(-1) were obtained at a hydraulic retention time of I h, and a nitrate loading of 0.24 kg N m(-3) d(-1). The process is extremely simple, stable and easy to maintain. PMID- 11905446 TI - Genetic parameters for antibody response of chickens to sheep red blood cells based on a selection experiment. AB - In the present study, a selection experiment for antibody (Ab) titer against SRBC in chickens was analyzed. Two lines were divergently selected for increased and decreased Ab titer. Further, a randombred control line, originating from the same base population, was included in the experiment. The heritability for immune response against SRBC was estimated after 18 generations of selection. In total, Ab titers obtained from 16,459 chickens were included in the analysis. Data was analyzed using an animal model. Posterior distributions for variance components and heritability were obtained using Markov Chain Monte Carlo methods. Response to selection was evaluated by constructing the posterior distribution for the genetic trend. In addition, a bivariate animal model was used in which male and female antibody titers were treated as different traits to estimate correlations between male and female immune response against SRBC. The heritability of Ab titer response, when using information from all three lines, was 0.18. The 90% highest posterior density region for the estimated heritability ranged from 0.16 to 0.19. In Generation 18, the genetic difference between the high and the low line was 5.1 phenotypic standard deviations. Analyses of each line separately revealed large differences in heritabilities between the lines, which could be mainly attributed to differences in error variances between the lines. The results suggest that selection for high Ab titers cause an increased environmental sensitivity. The estimated genetic correlation between male and female Ab titer was 0.92 and was not significantly different from 1. PMID- 11905447 TI - Biochemical characteristics of guanine nucleotide binding protein alpha-subunit recombinant protein and three mutants: investigation of a domain motion involved in GDP-GTP exchange. AB - Our previous studies using molecular dynamics have shown a hinge bending motion between the helical and the GTPase domains of GalphaT (Mello et al., 1998). The hypothesis that this motion is allowed by residues Gly56 and Gly179 and that this motion may affect the ligand exchange was tested in this work. Mutations of Gly 56 were carried out and the mutant proteins were expressed in Sf9 cells using the Baculovirus expression system. The recombinant proteins were purified using Ni NTA affinity chromatography. The results for the (GDP/GTP) exchange assays showed that G56S and double mutants (D55G/G56S) proteins differ significantly from the wild type and D55G mutant forms. The Kd values for GTPgammaS binding of those mutants have decreased by approximately 10-fold. No difference in the GTPase activity was detected for the mutants. Thus, the biochemical results obtained support the conclusions of the computational studies. PMID- 11905448 TI - Dental auscultation for nursing personnel as a model of oral health care education: development, baseline, and 6-month follow-up assessments. AB - Oral health care has been shown to have low priority in nursing and has been only partly successful. To create more positive effects than those achieved through traditional oral health care education, this project tested an educational model for nursing staff personnel. In addition to traditional oral health care education, some of the nursing staff members passed an additional dental auscultation period and served as oral care aides. The aides were responsible for the oral health care of the residents at their nursing facilities (intervention group). The intervention nursing facilities were compared with facilities where nursing personnel only received a traditional oral health care education program. Assessments were made at baseline and at a 6-month follow-up. At follow-up it was shown that the nursing staff in the intervention group gave higher priority to the oral health care work than the nursing staff in the control group. PMID- 11905449 TI - Diabetic foot ulcers and infections: current concepts. PMID- 11905450 TI - Understanding the payment system for hospital-owned wound care clinics. PMID- 11905451 TI - Arginine and wound healing. PMID- 11905452 TI - Management of a diabetic foot ulcer. PMID- 11905453 TI - Transcutaneous oximetry and skin surface temperature as objective measures of pressure ulcer risk. AB - OBJECTIVE: To determine if transcutaneous oximetry and skin surface temperature could measure differences in skin perfusion between 2 groups of hospitalized patients: (1) those who made gross or subtle movements in bed, and (2) those who could not move. DESIGN: A quasi-experimental design was used. SETTINGS AND PATIENTS: A convenience sample of 38 subjects (17 males, 21 females) was selected from patients admitted to a neurological/neurosurgical critical care and step down unit in a university health sciences center hospital. MAIN OUTCOME MEASURES: Prepressure and postpressure oximetry and temperature readings were taken over the sacrum and recorded every minute for 15 minutes. Pressure was applied by positioning subjects on their backs for 2 hours; subjects were observed for gross or subtle movement. Oximetry measurements were taken and recorded every minute. RESULTS: Paired t tests showed subjects who moved (n = 24) had a statistically significant difference between prepressure and postpressure temperature means over subjects who could not move (n = 14). Postpressure transcutaneous carbon dioxide (TcPCO2) mean levels were lower at all time-points and transcutaneous oxygen (TcPO2) mean levels were higher at most time-points in subjects who moved. Analysis of variance between the 2 groups during pressure showed a statistically significant difference in TcPCO2 levels, but not TcPO2 levels. CONCLUSIONS: Both methods identified statistically significant differences in moving and nonmoving patients and may prove to be useful tools in assessing pressure ulcer risk. Further research with both methods across various patient populations is needed. PMID- 11905454 TI - Guidelines in practice: the effect on healing of venous ulcers. AB - OBJECTIVE: To determine the effect of guidelines on vascular assessment, compression usage, dressing selection, and healing rates of patients with a venous ulcer. DESIGN: Prospective descriptive intervention evaluation. SETTING: Oxfordshire Community National Health Service (NHS) Trust, United Kingdom. PATIENTS: 40 consecutive prospective patients seen by Oxfordshire district nurses, either at home or at a wound clinic coordinated by district nurses located in a surgery office. INTERVENTION: The Guideline for Diagnosis and Treatment of Venous Leg Ulcers (University of Pennsylvania) and the Oxfordshire Leg Ulcer Guideline. MAIN OUTCOME MEASURES: Time to healing, compliance with vascular assessment, compression usage, and nursing costs. MAIN RESULTS: 91% of patients had a vascular assessment; all patients were treated with compression. Mean time to healing was 8 weeks and was not related to dressing selection or type of compression (short- versus long-stretch bandage). Nursing costs were slightly higher for wounds that healed after 12 weeks and were treated with a long-stretch bandage (Pound Sterling 170.00 [$250.00] vs Pound Sterling 272.00 [$395.00]). CONCLUSION: Use of compression was influenced by guidelines that emphasize a vascular assessment. Choice of dressing or type of compression was not a significant factor in healing rates. PMID- 11905456 TI - The point of education in wound care. PMID- 11905455 TI - Using skin replacement products to treat burns and wounds. AB - Much progress has been made toward the development of artificial skin replacement products. Continued research promises to bring more products to the marketplace, and each new product seems to develop a niche in the field of skin replacement. However, although each skin replacement product has unique properties and advantages, nothing works as well as a patient's own skin. Clinicians can only hope for an off-the-shelf skin replacement product that can be applied to a wound and yield a permanent, dependable dermis and epidermal skin replacement for all patients. PMID- 11905457 TI - Ipsilateral local recurrence of breast cancer: determinant or indicator of poor prognosis? AB - The importance of ipsilateral local recurrence within a conserved breast depends on the micrometastatic environment at the time of initial clinical presentation. In the absence of micrometastases, local recurrence would be a determinant of distant disease; however, in the presence of micrometastases, it represents a marker of distant relapse. Maximum locoregional treatment at primary diagnosis would be appropriate in the former group, whereas minimum treatment would be sufficient in the latter group, with full treatment prescribed at the time of local recurrence. As an indicator of poor prognosis, the presence of local recurrence permits a more selective approach to therapies that would otherwise result in overtreatment for some patients. We, therefore propose a trial that compares conventional treatment with minimum therapy at presentation plus maximum therapy at local relapse in postmenopausal women with small tumours. PMID- 11905458 TI - Clinical role of sentinel-lymph-node biopsy in breast cancer. AB - The introduction of sentinel-lymph-node biopsy has brought new impetus to the early staging of cancer in general, and breast cancer in particular. This technique has rekindled the discussion on the present role and routine practice of axillary-lymph-node dissection in early breast cancer, the methods available for the histopathological assessment of lymph nodes, and the current thoughts about best surgical practice in the management of breast cancer. Sentinel-lymph node biopsy has spread so rapidly that surgeons, pathologists, and patients are no longer willing or able to ignore the possible consequences of its implementation. A vast amount of data (over 1150 publications in the peer reviewed literature on this subject to date) attests to the explosive interest in the past 5 years. In this article we review our own experience and discuss recommendations for clinical practice. PMID- 11905459 TI - Emerging pathways in colorectal-cancer development. AB - The past decade has seen the emergence of new pathways in the development of colorectal cancer. There is now clear evidence that subsets of these tumours do not show chromosomal instability and do not follow the suppressor pathway. Instead, about 15% of colorectal cancers are characterised by microsatellite instability (MSI). This feature arises through defective DNA mismatch repair, which is related either to a germline mutation (as in hereditary non-polyposis colorectal carcinoma) or to failure to express a mismatch-repair gene. CpG-island methylation has been linked to sporadic cancers with a high frequency of MSI. This type of methylation leads to loss of gene expression when it occurs in the promoter region of a gene. Tumours may have high or low type C (cancer-related) CpG-island methylation. When methylation affects hMLH1 (mismatch repair gene), the resultant cancer has high MSI. PMID- 11905460 TI - Shortage of varicella and measles, mumps and rubella vaccines and interim recommendations from the Advisory Committee on Immunization Practices. PMID- 11905462 TI - Committed to kids: an integrated, 4-level team approach to weight management in adolescents. AB - The integrated, 4-level approach of Committed to Kids is successful because of several factors: The sessions are designed to entertain the adolescents and promote initial success; The program features parent-training methods in short, interactive, educational sessions; In severely obese adolescents, the diet intervention results in noticeable weight loss that motivates the patient to continue; also, the improved exercise tolerance resulting from the weight loss promotes increased physical activity; and The program team provides consistent feedback-patients and their families receive results and updates every 3 months. Most importantly, the program is conducted in groups of families. The adolescent group dynamics and peer modeling are primary components of the successful management of obesity in youth. PMID- 11905463 TI - cAMP response element-binding protein interacts with the homeodomain protein Cdx2 and enhances transcriptional activity. PMID- 11905461 TI - Low accuracy and low consistency of fourth-graders' school breakfast and school lunch recalls. AB - OBJECTIVE: To determine the accuracy and consistency of fourth-graders' school breakfast and school lunch recalls obtained during 24-hour recalls and compared with observed intake. DESIGN: Children were interviewed using a multiple-pass protocol at school the morning after being observed eating school breakfast and school lunch. SUBJECTS: 104 children stratified by ethnicity (African-American, white) and gender were randomly selected and interviewed up to 3 times each with 4 to 14 weeks between each interview. STATISTICAL ANALYSIS: Match, omission, and intrusion rates to determine accuracy of reporting items; arithmetic and/or absolute differences to determine accuracy for reporting amounts; total inaccuracy to determine inaccuracy for reporting items and amounts combined; intraclass correlation coefficients (ICC) to determine consistency. RESULTS: Means were 51% for omission rate, 39% for intrusion rate, and 7.1 servings for total inaccuracy. Total inaccuracy decreased significantly from the first to the third recall (P=0.006). The ICC was 0.29 for total inaccuracy and 0.15 for omission rate. For all meal components except bread/grain and beverage, there were more omissions than intrusions. Mean arithmetic and absolute differences per serving in amount reported for matches were -0.08 and 0.24, respectively. Mean amounts per serving of omissions and intrusions were 0.86 and 0.80, respectively. APPLICATIONS/CONCLUSIONS: The low accuracy and low consistency of children's recalls from this study raise concerns regarding the current uses of dietary recalls obtained from children. To improve the accuracy and consistency of children's dietary recalls, validation studies are needed to determine the best way(s) to interview children. PMID- 11905465 TI - The 2000 Nobel Prize in physiology or medicine. PMID- 11905466 TI - Scientists and society--and never the twain shall meet? PMID- 11905464 TI - Chemistry meets biology. PMID- 11905467 TI - Bioethics--an outlook. PMID- 11905468 TI - Bioethics and epistemic-moral hybrids: perspectives from the history of science. PMID- 11905469 TI - Reflecting bioethics. PMID- 11905470 TI - Experts on philosophical reflection in public discourse--the German Sloterdijk debate as an example. PMID- 11905471 TI - With a different voice? New French contributions to international bioethics. PMID- 11905472 TI - Biomedical ethics in South Africa: current challenges and institutional responses. PMID- 11905473 TI - Notes on bioethics in Brazil. PMID- 11905474 TI - Bioethics and medical practice in the age of molecular genetics. PMID- 11905475 TI - Revising the Declaration of Helsinki. PMID- 11905476 TI - The Declaration of Monaco. AB - The International Symposium on Bioethics and the Rights of the Child was held in Monaco at the end of April, jointly organized by the World Association of Children's Friends (AMADE) and UNESCO. At the end of the meeting the following declaration was agreed with a view to reinforcing the protection of children's rights. PMID- 11905477 TI - Human genetic improvement: a comparison of Russian and British public perceptions. PMID- 11905478 TI - Europe moves to ban human cloning. PMID- 11905479 TI - Declaration of Hamburg. AB - At its annual meeting in Hamburg, Germany, at the end of last year the World Medical Association adopted several declarations and other statements, two of which are reprinted here. PMID- 11905481 TI - Populations receiving optimally fluoridated public drinking water--United States, 2000. AB - Dental caries (i.e., tooth decay) is a transmissible, multifactor disease that affects 50% of children aged 5-9 years, 67% of adolescents aged 12-17 years, and 94% of adults aged > or = 18 years in the United States. During the second half of the 20th century, a major decline in the prevalence and severity of dental caries resulted from the identification of fluoride as an effective method of preventing caries. Fluoridation of the public water supply is the most equitable, cost-effective, and cost-saving method of delivering fluoride to the community. In the United States during 2000, approximately 162 million persons (65.8% of the population served by public water systems) received optimally fluoridated water compared with 144 million (62.1%) in 1992. This report presents state-specific data on the status of water fluoridation in the United States and describes a new surveillance system designed to routinely produce state and national data to monitor fluoridation in the public water supply. The results of this report indicate slow progress toward increasing access to optimally fluoridated water for persons using public water systems. Data from the new surveillance system can heighten public awareness of this effective caries prevention measure and can be used to identify areas where additional health promotion efforts are needed. PMID- 11905480 TI - Laboratory-acquired meningococcal disease--United States, 2000. AB - Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis among older children and young adults in the United States. N. meningitidis usually is transmitted through close contact with aerosols or secretions from the human nasopharynx. Although N. meningitidis is regularly isolated in clinical laboratories, it has infrequently been reported as a cause of laboratory-acquired infection. This report describes two probable cases of fatal laboratory-acquired meningococcal disease and the results of an inquiry to identify previously unreported cases. The findings indicate that N. meningitidis isolates pose a risk for microbiologists and should be handled in a manner that minimizes risk for exposure to aerosols or droplets. PMID- 11905482 TI - Socioeconomic status of women with diabetes--United States, 2000. AB - Persons whose socioeconomic status is low have poorer health than other persons and are less likely to have adequate access to care or to receive high-quality clinical and prevention care services. In the United States, diabetes is a potentially debilitating disease that is increasing in prevalence; however, little is known about the socioeconomic status of persons with diabetes. Women account for approximately 52% of all persons aged > or = 20 years with diabetes. To assess the socioeconomic status of women with diabetes, CDC analyzed data from the Behavioral Risk Factor Surveillance System (BRFSS), which indicated that the socioeconomic status of women with diabetes in 2000 was markedly lower than that of women without diabetes. Efforts should be focused to understand the impact of socioeconomic conditions on the health and quality of care of women with diabetes. PMID- 11905483 TI - The association of childhood asthma with parental employment and welfare receipt. AB - OBJECTIVE: to assess the association of childhood asthma with parental employment and welfare receipt, which has not been studied. METHOD: We analyzed cross sectional data on 13 371 children younger than 18 from the 1997 National Health Interview Survey, a nationally representative stratified probability sample of the US noninstitutionalized population. Single-parent (n=3,907) and 2-parent families (n=9,464) were analyzed separately. Families with children younger than 6 and families with incomes below the federal poverty level (FPL) were also analyzed separately. The main outcome measures were full-time parental employment and welfare receipt. RESULTS: Compared to single parents of nonasthmatic children younger than 6, single parents of young children with asthma were more likely to be employed less than full time (adjusted odds ratio [OR] 2.1, 95% confidence interval [CI], 1.4-3.2). This relationship was also evident among single-parent families with incomes below FPL (adjusted OR 2.8, 95% CI, 1.2-6.5). Parental employment among 2-parent families with young children was similar regardless of child's asthma status. Two-parent families with young asthmatic children were more likely to have received welfare for at least 1 parent (adjusted OR 2.6, 95% CI, 1.5-4.6). Single-parent families of asthmatic children were also more likely to have received welfare (adjusted OR 1.4, 95% CI, 1.1-1.7). CONCLUSION: Children's asthma is associated with reduced parental employment among single parents and increased welfare receipt among single- and 2-parent families. These associations with children's asthma may have implications for policy makers interested in increasing employment and decreasing welfare PMID- 11905485 TI - Leaving welfare often severs families' connections to the Food Stamp Program. AB - In the early stages of welfare reform, a variety of studies found that many families that left welfare also left the Food Stamp Program (FSP), even though most were still eligible for it. This paper examines more recent data to determine whether this behavior has continued or whether at least some of it was due to initial misunderstandings about eligibility. Our results show that families leaving welfare continued leaving the FSP at about the same rate in 1999 as they had 2 years earlier. For example, only half of families who left welfare and had incomes below 50% of the federal poverty level continued participating in the FSP. We also show that more families reported leaving the FSP because of administrative issues in 1999 than did in 1997. Families that left the FSP were more likely to have moved recently and to have owned cars than families that remained. Our results raise concern that structural reforms are needed to make it easier for the working poor to receive these benefits. Recent federal changes in the FSP have allowed states to move in that direction. PMID- 11905484 TI - Five years later: poor women's health care coverage after welfare reform. AB - The 1996 welfare reform law aims to increase poor women's participation in the work force and encourage their financial independence. Because women's ability to obtain and retain employment is affected by their health status, welfare reform's success is fundamentally tied to poor women's access to health care and to health insurance. Despite this, the rate of uninsurance among poor women with children has grown by half in recent years, leaving 37% of poor mothers uninsured in 2000. Coverage through employer-sponsored insurance has increased only slightly, and Medicaid participation has dropped. Although many factors contributed to this, welfare policies and procedures and low Medicaid eligibility levels had unintended yet significant negative effects on women's health care coverage. The sharp decline in poor women's health care coverage is likely to be one of several health-related issues that Congress will consider as it debates the reauthorization of the welfare law in 2002. Both public and private efforts will be necessary to improve coverage for poor women with children. Much progress has been made during the past 5 years in covering poor and near-poor children, but their parents have been left behind. The same efforts that proved successful for children, including broadening eligibility for coverage and simplifying the application process, can be used to improve the health and well-being of parents and to strengthen their ability to care for and support their families. PMID- 11905486 TI - Depression among women on welfare: a review of the literature. AB - In the "work-first" welfare policy environment embodied in the Personal Responsibility and Work Opportunity Reconciliation Act of 1996, situational and personal barriers to employment have received increasing scrutiny from researchers, policy makers, and program administrators. Depression has generated a great deal of concern as a potential barrier to job attainment and retention among welfare recipients. This paper reviews the literature on the prevalence of depression in recent studies of welfare recipients. Starting in the late 1980s, many researchers incorporated reliable and comparable measures of major depression and depressive symptoms into their studies of welfare recipients. These investigations show that rates of depression are high, as is the presence of symptoms. The relevance of this problem for welfare administrators and other professionals is discussed. PMID- 11905487 TI - Welfare, women, and children: it's time for doctors to speak out. PMID- 11905488 TI - Keeping battered women safe during welfare reform: new challenges. AB - This paper reviews the growing body of research literature on the relationship of domestic violence to welfare. Not only do women on welfare suffer from domestic violence in far greater numbers than women in the general population, but their abusers, threatened by the women's efforts at education, training, or work, also use violence and threats of violence to sabotage these efforts at economic self sufficiency. For this reason, welfare reform exacerbates domestic violence in the lives of many low-income women. As a result of the federal Family Violence Option, most state welfare plans allow battered women on welfare more time and specialized services before mandating work in order to keep them and their children safe. Recent research and monitoring have shown, however, that the majority of battered women on welfare do not tell their welfare workers about the violence. Ending the isolation of these battered women and helping them with domestic violence services pose difficult challenges. Women's health providers may be in a better position to accomplish this task than welfare department personnel. PMID- 11905489 TI - Substance abuse and welfare reform. AB - The Personal Responsibility and Work Opportunity Reconciliation Act of 1996 has forced welfare officials and substance abuse treatment providers to better understand the connections between substance abuse and welfare dependency. As a result, both are thinking differently about their policies and even their missions. Little is known about the true extent of substance abuse among welfare recipients, but estimates range from 2.7% to 22%. The current system of substance abuse treatment is poorly matched both to the needs of disadvantaged mothers and to the mandates of welfare reform. To help women achieve self-sufficiency, treatment programs must focus on the needs of women, address problems that often accompany substance abuse, attend to the needs of children, and emphasize work as an outcome of treatment. Welfare agencies must also change to meet their mandates; they must improve the tools and resources they offer staff, improve access and services for families, and align agency policies with new missions. PMID- 11905490 TI - Abstinence-only programs implemented under welfare reform are incompatible with research on effective sexuality education. AB - Abstinence-only-until-marriage education programs receive more than $100 million annually in government funds, most of it stemming from the Personal Responsibility and Work Opportunity Reconciliation Act of 1996. There is currently no evidence that any of these programs, which promote sexual abstinence and restrict information about contraception, actually achieve their intended purposes. On the other hand, there is ample evidence that comprehensive sexuality education programs, which include information about both abstinence and contraception, can be effective. We argue that public monies should be expended on programs with proven efficacy and not on those that promote ideologies that are not shared by the majority of Americans. PMID- 11905491 TI - Overhauling welfare: implications for reproductive health policy in the United States. AB - The overhaul of the welfare system in 1996 broke the historic link between eligibility for welfare benefits and eligibility for Medicaid, ended a longstanding requirement that welfare recipients be given access to family planning services, and, at the same time, included a number of controversial features directed at reducing out-of-wedlock childbearing and promoting abstinence-only education. Five years later, the number of women enrolled in Medicaid is down, the number not covered by insurance is up, and abstinence-only education has become a prominent feature of the government's effort to decrease the incidence of out-of-wedlock pregnancies. Very little is known about how well these interventions are working and what impact, positive or negative, they have had. As Congress heads for reauthorization of the nation's welfare law later this year, policy makers should give increased attention to the provision of information and services, including those related to family planning, in order to better address the reproductive health needs of low-income women. PMID- 11905492 TI - Maternity rights, work, and health in France and Italy. AB - This article focuses on the principles and the implementation of maternity rights (MR) in France and Italy. Results show that MR are well established in both countries, where about 80% of women employed during pregnancy were back to work 1 year after childbirth. Nevertheless, social inequalities were found. Less educated women and those who had manual jobs or worked in small firms in the private sector or off-the-books were less likely to take an extended leave and to return to work. Despite differences in child care provisions, quality and accessibility of child care were common concerns for both French and Italian mothers. Employment was not related to any health problem in Italy 1 year after birth; in France, unemployed new mothers had high rates of psychological distress. Financial worries and marital problems were associated with several health problems in both countries. In conclusion, combining work and motherhood is possible in these 2 countries without too many costs for women, at least for the more privileged among them. However, this relative ease could vanish if social and economic conditions changed for the worse. PMID- 11905493 TI - Welfare reforms in Australia: how will they affect women's health? AB - Like most Western countries, Australia is in the process of introducing welfare reforms to curb costs. Australian reforms follow and are informed by similar reforms in the United States and United Kingdom and will be incrementally implemented until 2003. Australian reforms emphasize mutual obligation, preventing people from "taking advantage" of the welfare system, and avoiding long-term reliance on welfare. In contrast to the United States, where the mothers of young children have specifically been targeted, reforms in Australia do not privilege women's roles as workers over their roles as caregivers. Work obligations will be introduced only for mothers whose youngest child is older than 16 years. In fact, financial incentives for providing care for young children and people with disabilities have actually increased. Existing health research suggests that the impact of welfare reform on both health and society will depend on how the balance between women's roles as caregivers and workers is struck. PMID- 11905494 TI - What causes job loss among former welfare recipients: the role of family health problems. AB - OBJECTIVE: to test whether women's or children's health status influences the likelihood that low-income single mothers experience job loss. METHODS: Using a nationally representative probability sample from the National Longitudinal Survey of Youth, we estimated whether having a health limitation or having a child with a health limitation was associated with job loss for a sample of 783 women who had previously been on welfare. RESULTS: Both having a health limitation (odds ratio [OR]=1.53; 95% confidence interval [CI], 1.19-1.97) and having a child with a health limitation (OR=1.36; 95% CI, 1.18-1.56) were associated with significantly increased risk of job loss among women previously on welfare. The effects remained significant after adjustment for age, education, marital status, race, age and number of children, and economic conditions. CONCLUSIONS: Dramatic changes in welfare policy in the United States have made many single mothers living in poverty dependent on work as their sole source of income. Although studies have shown that families on welfare are more likely to have health limitations, little is known about how family health affects the ability of poor single mothers to remain employed. These results demonstrate that women with health limitations and mothers of children with health limitations are at particularly high risk of losing their jobs. Public and private policies that can help reduce job loss as a consequence of family health problems are discussed. PMID- 11905496 TI - Surgical research funding. PMID- 11905495 TI - Welfare, women, and families: implications for clinicians. AB - Although the 1996 welfare reform law signaled a most profound shift in US social policy, clinicians do not fully appreciate the potential impact this legislation could have on patterns of health and the provision of health services to millions of American women and children. The data presented in this issue of JAMWA point to definitive steps clinicians can take to provide optimal care for their patients. First, we must commit to educating ourselves and others who care for patients about the nature and potential impact of welfare reform. Second, we must devise efficient and effective ways of identifying and addressing these needs in our clinical settings. Third, clinicians should use their experience to effectively advocate for their patients on individual and population levels. As clinicians, we have no choice but to respond to the social forces that so profoundly affect the health of the families we serve; we must take advantage of our capacity to make substantial contributions to the health and well-being of our patients. PMID- 11905497 TI - Founders of modern surgery. PMID- 11905498 TI - Warm hepatic ischemia in pigs: effects of L-arginine and oligotide treatment. AB - Reperfusion injury represents a key event leading to graft nonfunction. Maintaining adequate nitric oxide levels and stimulating vasodilator synthesis can probably minimize endothelial damage. The aim of this study was to investigate the effect of L-arginine, a substrate of nitric oxide synthesis, and oligotide, a promoter of prostacyclin synthesis, on liver function and morphology after warm ischemia-reperfusion injury. After constructing a side-to-side portacaval shunt, ischemia was induced by clamping the hepatic hilum for 2 h above the shunt, in 19 female pigs divided into a control group (n = 7), an L arginine treatment group (n = 6), and an oligotide treatment group (n = 6). Liver function tests and measurements of serum and red blood cell malondialdehyde and plasma nitric oxide levels were performed before reperfusion and at 1, 10, 60, and 120 min after reperfusion. Liver biopsies, taken before reperfusion and at 30 min and 7 days after reperfusion, were analyzed for tissue malondialdehyde, histological-ultrastructural features, and apoptosis evaluation. Thirty minutes after reperfusion, liver malondialdehyde, sinusoidal congestion, necrosis, and apoptosis were significantly lower in the L-arginine group than in the controls (p < .05). On postoperative day 7, tissue malondialdehyde decreased, while plasma nitric oxide and hepatocyte glycogen content were increased in the L-arginine group compared to controls (p < .05). This study demonstrates the protective effect of L-arginine on hepatic lipoperoxidation and liver morphology in a pig model of warm ischemia-reperfusion injury. The increased plasma levels of nitric oxide a week after ischemia-reperfusion injury support the hypothesis that it has a role in preventing liver damage. The same beneficial effect was not confirmed for oligotide. PMID- 11905499 TI - Exogenous nitric oxide protects kidney from ischemia/reperfusion. AB - Blockade of NO production is followed by an increase in leukocyte rolling and adhesion resulting in some deleterious effects of ischemia. Preischemic administration of NO protects vascular integrity after reperfusion. Exogenous NO causes a direct reduction in leukocyte adhesion. This work was performed to test the hypothesis that exogenous NO administered during the preischemic period to the kidney alone, without coming into contact with the leukocytes, could also reduce leukocyte-endothelium adhesion. Adult rats were subjected to in situ isolation of the left kidney. Solutions were infused through the renal artery and drained through an incision in the renal vein, thus avoiding the systemic circulation. Group IC rats served as an ischemic control, and received 0.9% saline. Group NP received Na nitroprusside. Group S was a nonischemic control. Groups IC and NP were subjected to 75 min of renal ischemia. After this period, vascular structures were repaired and reperfusion allowed. A right nephrectomy was performed. Serum urea and creatinine, myeloperoxidase activity, and histopathological studies were carried out at different intervals after reperfusion. Survival at 15 days was 46%, 80%, and 100% in groups IC, NP, and S, respectively. Differences between groups for serum urea and creatinine were significant only during the first seven days. Myeloperoxidase values were significantly higher in group IC. All rats from group IC and only 20% from group NP showed histological evidence of necrosis. Thus, exogenous NO is protective and acts selectively upon the kidney, modulating its interactions with polymorphonuclear cells after ischemia/reperfusion. PMID- 11905500 TI - Evaluation of a xenogeneic acellular collagen matrix as a small-diameter vascular graft in dogs--preliminary observations. AB - Autogenous veins are the materials of choice for arterial reconstruction. In the absence of autogenous material, prosthetic materials are used. However, vascular prostheses of less than 0.4 cm in diameter have low long-term patency. This study was designed to determine if cells would infiltrate an engineered xenogeneic biomaterial used as a small diameter arterial graft in dogs and, if so, to determine the phenotype of the infiltrating cells. Nine acellular xenogeneic grafts (0.4 cm in diameter, 5 cm long), composed of porcine collagen derived from the submucosa of the small intestine and type I bovine collagen, were implanted as end to-end interposition grafts in femoral arteries of five male mongrel dogs (total of nine grafts). All dogs received daily aspirin (325 mg). Patency of implanted grafts was monitored weekly by Duplex ultrasonography. After 9 weeks, or earlier in case of blood flow reduction by at least 75%, grafts were explanted and prepared for light or electron microscopy to evaluate cellularization. Eight of nine grafts remained patent up to 9 weeks. At explant, diameters were 0.31 +/- 0.02 cm at the midgraft, and 0.14 +/- 0.01 and 0.19 +/- 0.01 cm at the proximal and distal anastomoses. At explant, cells of mesenchymal origin (endothelial cells, smooth muscle cells, myofibroblasts) were embedded in the extracellular matrix of the graft scaffold. Minimal evidence of cellular inflammatory reaction and no aneurysmal dilatation or thrombus formation was detected. Variable degrees of hyperplasia were present at proximal and distal anastomoses. This preliminary study demonstrates that a collagen-based xenogeneic biomaterial provides a scaffold for cellularization when used for arterial reconstruction in dogs. PMID- 11905501 TI - Effect of perfusion and preservation on IL1-beta, IL-6, and IL-10 production in murine lung. AB - We evaluated the production of the interleukins (ILs) IL-1beta, IL-6, and IL-10 in both the vasculature and pulmonary tissue before and after 24 h of lung preservation. The cardiopulmonary blocs of 21 Balb-c mice were divided into three study groups (7 mice/group) and were flushed through the pulmonary artery with Krebs-Henseleit buffer (K-Hb) at 4 degrees C at a rate of 0.2 mL/min as follows: Group 1, lung washout: lungs were flushed until pulmonary effluent was clear. Group 2, perfusion: After lungs were flushed until pulmonary effluent was clear, lungs were perfused during 30 min. Group 3, preservation: Lungs were flushed until pulmonary effluent was clear, and the cardiopulmonary bloc was preserved immersed into (K-Hb) at 4 degrees C. After 24 h of preservation, lungs were reperfused during 30 min. In all study groups the caudal lobe from the left lung was taken for microscopical study; all other lobes were homogenized with (K-Hb) and the supernatant was obtained. IL-1beta, IL-6, and IL-10 production in lung effluents (washout, perfusion, and reperfusion) and in lung tissue were measured by enzyme-linked immunosorbent assay (ELISA). In the lung effluent, there was no statistical difference between IL-1beta and IL-6 concentrations. In all study groups, IL-10 production was significantly higher than IL-1beta and IL-6 levels. IL-10 level was lowest in the 24-h preservation group when it was compared to the other groups. In group 1, there was a negative correlation (r = -.599, p < .05) between IL-1beta and IL-10. In pulmonary tissue, IL-1beta was higher in group 2 when compared to groups 1 (p = .001) and 3 (p = .002), and it was significantly lower in group 3. IL-10 was lower in group 1 when compared to groups 2 (p = .001) and 3 (p = .004). In groups 1 and 2, IL-1beta was significantly higher than IL-6 and IL-10. In group 3, IL-10 was higher than IL-1beta (p = .0001) and IL-6 (p = .0001). Correlation of effluent/tissue index with histological findings showed a negative correlation between IL-10 effluent/tissue relation and inflammation (r = -.68, p < .01). In conclusion, the main cytokine found in lung effluents was IL 10, followed by IL-6 and IL-1beta. On the other hand, cytokine concentration in lung tissue homogenates was mainly due to the presence of IL-1beta. However, this cytokine shows a significant reduction in lung tissue after prolonged preservation. PMID- 11905502 TI - Validation of conductance catheter system for quantification of murine pressure volume loops. AB - The purpose of this study was to define the validation methods and outcomes of a conductance catheter system specifically for in vivo murine cardiac hemodynamic analysis. To express the relationship between conductance and blood volumes, we used an in vitro model to derive a volume-conductance line. The volume conductance line was used to compute raw volume from the modified conductance signals. The parallel volume was calibrated with hypertonic (15%) saline injected from extrajugular vein. The ventricular volume was computed by raw volume minus parallel volume. The accuracy of conductance volumetric measurements was validated with a static in situ infusion of calibrated volumes of whole blood injected into arrested left ventricles. In vivo dynamic measurements were performed with 24 C57B1/6 mice, 6 months old; for comparison of established values. The in situ model showed that after calibration, the experimental coefficient, alpha, was equal to 1 and the measured volume by conductance catheter was equal to the true volume of the left ventricle (y = 0.982x + 0.513, p < .0001). For the in vivo models, the end-diastolic volumes and the stroke volumes and cardiac output determined with the conductance catheter system were 17.3 +/- 1.0 microL, 10.6 +/- 0.9 microL, and 6.0 +/- 0.5 mL/min, respectively. We validated the relationship between measured volume by conductance catheter and the true volume and demonstrated the accuracy of the volume-conductance line for conversion of conductance to volume. PMID- 11905503 TI - Cooled-tip diode laser catheter for improved catheter ablation of ventricular tachycardia. AB - Catheter ablation for the treatment of arrhythmias has evolved dramatically over the past two decades. Researchers have investigated alternative energy sources and catheter constructs to improve the efficacy and safety of catheter ablation. This study tested the hypothesis that a new prototype cooled-tip laser catheter used with a low-power diode laser would improve catheter ablation of ventricular tachycardias. Four mongrel dogs underwent a median sternotomy. The cooled-tip laser catheter was advanced into the left ventricle via the left carotid artery and positioned adjacent to the endocardium. Laser powers of 3 and 4 W were delivered at four exposure times to select areas of the endocardium. During application of laser energy, room-temperature saline was circulated through the catheter. At necropsy the hearts were examined and fixed in formalin for histologic examination. Gross examination of the endocardial surfaces showed no indication of crater or thrombus formation. Cross-section of the lesions revealed sharply demarcated, circular-shaped areas of coagulative necrosis extending into the mid-myocardium. Areas of coagulative necrosis were identified within the myocardium extending into the mid-myocardium and occasionally the subepicardium. A sharp line of demarcation was observed between the lesions and the surrounding normal myocardium. The results of this study showed that we could use surface cooling during slow laser heating to create large subsurface lesions with characteristics appropriate for treatment of ventricular tachycardia and little to no surface damage. We believe our catheter system addresses many of the previous issues with laser-based approaches. PMID- 11905504 TI - Thalidomide, a drug which may protect the organism against some aggressive effects of the immunologic system. PMID- 11905505 TI - Management of erythema nodosum leprosum by thalidomide: thalidomide analogues inhibit M. leprae-induced TNFalpha production in vitro. AB - Thalidomide is being successfully used for the treatment of erythema nodosum leprosum (ENL), among other disorders with inflammatory and immunological bases. Although the active molecules responsible for the diverse therapeutic activities of the drug and the sequence of reactions triggered inside the cells remain unclear, it was demonstrated that thalidomide (THAL) inhibits TNFalpha mRNA expression and protein production by stimulated monocytes and activated T lymphocytes. Patients treated with THAL experienced a reduction in serum TNFalpha levels and it diminished cytokine gene expression at the lesion site, with a concomitant abrogation of clinical symptoms. It has been reported that thalidomide as well as some its analogues decrease M. leprae-induced TNFalpha and IL-12 mRNA in vitro. THAL also reduced monocyte apoptosis in the cultures. The present data further support thalidomide's effects on TNFa synthesis and the growing need to search for new specific TNFalpha inhibitors (non-teratogenic compounds) that might be potentially used in clinical disorders such as leprosy. PMID- 11905506 TI - Thalidomide in myelodysplastic syndromes. AB - The myelodysplastic syndromes are a heterogeneous group of clonal diseases of haemopoiesis, which are a challenge for both biologists and clinicians. In this paper the current classification and the recent advances in the understanding the disease mechanisms are reviewed. The recent therapeutic advances are also indicated, such as intensive and low-dose chemotherapy, new drugs, erythropoietin and colony-stimulating factors. However, the work has been focused on thalidomide, its therapeutic potential, its modes of actions, side effects, indications and future applications. PMID- 11905507 TI - Estrogens and environmental estrogens. AB - The natural female sex hormone estrogens binds once inside the cell to a protein receptor to form a 'ligand-hormone receptor complex'. The binding activates the hormone receptor, which triggers specific cellular processes. The activated hormone receptor then turns on specific genes, causing cellular changes that lead to responses typical of a ligand-hormone receptor complex. Estrogens (especially estradiol) bring out the feminine characteristics, control reproductive cycles and pregnancy, influence skin, bone, the cardiovascular system and immunity. Natural hormones are more potent than any of the known synthetic environmental estrogens (except drugs such as diethylstilbestrol [DES]). Estrogen production varies according to different factors (gender, age and reproductive cycles). Women produce more estrogen than men and the production is more abundant during fetal development than in the postmenopausal period. Most natural estrogens are short-lived, do not accumulate in tissue and are easily broken down in the liver. In contrast to natural estrogens, estrogenic drugs such as ethynylestradiol diethylstilbestrol (DES), synthetic environmental estrogens such as beta hexachlorocyclohexane (beta-HCH), polychlorinated biphenyls (PCBs), o, p, p'DDT, 4-nonylphenol (NP) and phytoestrogens such as isoflavones or lignans, are more stable and remain in the body longer than natural estrogens. Because most of these compounds are lipophilic, they tend to accumulate within the fat and tissue of animals and humans. Thus, depending on the natural estrogen levels, environmental estrogens may have different influences (mimicking, blocking or cancelling out estrogen's effects) on estrogen activities. PMID- 11905508 TI - Thalidomide in cancer. AB - Thalidomide has immunomodulatory and anti-angiogenic properties which may underlie its activity in cancer. After its success in myeloma, it has been investigated in other plasma cell dyscrasias, myelodysplastic syndromes, gliomas, Kaposi's sarcoma, renal cell carcinoma, advanced breast cancer, and colon cancer. Thalidomide causes responses in 30-50% of myeloma patients as a single agent, and acts synergistically with corticosteroids and chemotherapy. Thalidomide results in the reduction or elimination of transfusion-dependence in some patients with myelodysplastic syndrome. Responses have also been seen in one-third of patients with Kaposi's sarcoma, in a small proportion of patients with renal cell carcinoma and high-grade glioma, and in some patients with colon cancer in combination with irinotecan. The drug is being investigated currently in a number of clinical trials for cancer. Drowsiness, constipation, and fatigue are common side effects, whereas peripheral neuropathy and skin rash are seen in one-third. A minority of patients experience bradycardia. Thrombotic phenomena are especially common when thalidomide is combined with chemotherapy. Adverse effects severe enough to necessitate cessation of therapy are seen in around 20% of patients. A therapeutic trial of thalidomide is essential in all patients with relapsed or refractory myeloma. In other cancers, the best way to use the drug is in the setting of clinical trials. In the absence of access to studies or alternative therapeutic options, thalidomide could be considered singly or in combination with standard therapy. PMID- 11905509 TI - Auditory adaptation is differentially impaired in familial and sporadic Alzheimer's disease. AB - Neurophysiologic measures are particularly sensitive to alterations in attention and arousal. The purpose of this study was to evaluate the auditory adaptation of normal and mildly demented elderly people. We compared the automatic behavior of an auditory evoked potential (N100) in three age-matched groups of elderly subjects, one with familial Alzheimer's disease (AD), one with sporadic AD and one healthy group. All AD subjects corresponded clinically and neuropsychologically with the early stage of dementia. The dynamic range of auditory adaptation is known to be related to age, and normal auditory adaptation for the age was observed in our healthy aged and sporadic AD subjects, whereas the familial AD subjects lacked normal adaptation. The familial AD subjects also showed statistically significantly smaller peak amplitudes and shorter latencies of the N100 throughout the habituation test. This persistent difference in automatic habituation of sensory responses supports the view that different subtypes of AD are differentially affected. The observed differences give an objective measure of the impaired involuntary adaptive functions of neuronal networks involved in auditory processing in subtypes of AD. Since habituation reflects the most primitive stage of learning and short-term memory, altered habituation may predict faster deterioration of clinical status in the familial group of AD subjects. PMID- 11905510 TI - Interplays between genetic and environmental mechanisms trigger tumorigenic VEGF signalling in human HCC cell lines: pilot study. AB - The 'angiogenic switch' concept has been used to discover various pro- and anti angiogenic molecules as pharmacotherapeutic strategies for cancers and other ischemic and inflammatory diseases; however, surprisingly little is known about the 'tumor angiogenic switch' in response to complex interplay between environmental and genetic mechanisms that are most importantly, hypothesis driven, largely unsolved at the postgenomic level. The present study's aim is to identify those interplays between the expressing green fluorescence protein (EGFP) transcript and redox-driven vascular endothelial growth factor (VEGF) upstream mechanisms that influence tumorigenic VEGF signalling, by using multifactorial orthogonal statistical analyses, the non-transfected and transfected human hepatocellular carcinoma (HCC) cell lines, and quantitative 'sandwich' Elisa immunoassays. The unique results indicate valuable findings on the postgenomic level as follows. Fusion of the EGFP significantly triggers tumorigenic VEGF signalling in three E3-, F11- and A(3-1)-transfected human HCC cell lines, compared with the parental EGFP-free malignant hepatocellular carcinoma (MHCC)1 cell line; Redox-regulated VEGF upstream mechanism interplays dramatically mediate dual responses, either down- or upregulating tumorigenic VEGF signalling, which depends on individual post-transcriptional regulation or upstream modification of the VEGF promoter in the three MHCC1, SMCC7721 and doxorubicin-resistance 7402/D+ non-transfected human HCC cell lines; and mechanism-based strategies for stimulating beta-adrenergic and P2-purinergic signal transduction and counteracting O2 and Ca2+-inflow significantly trigger tumorigenic VEGF signalling in ATRA/ATRP-driven networks, compared with controls of the non-transfected cell types examined. The contrast data offer first pilot paradigms of searching for hypothesis-driven multifactorial interplays on tumorigenic VEGF signalling, which are valuable for further deciphering postgenome-wide VEGF upstream regulatory networks of switching on the tumor angiogenesis, and developing mechanism-based novel pharmacotherapeutic strategies. PMID- 11905511 TI - Lipid-lowering drugs (statins), cholesterol, and coenzyme Q10. The Baycol case--a modern Pandora's box. PMID- 11905512 TI - Effects of the infusion mode on bicarbonate balance in on-line hemodiafiltration. AB - BACKGROUND: Electrolyte and acid-base balance may be differently affected by the infusion mode in on-line hemodiafiltration (HDF). We studied the effects of the different infusion modes on bicarbonate transport across the dialyzer membrane, and thus on the final bicarbonate balance of the HDF sessions. METHODS: Instantaneous HCO3- transfer across the dialyzer membrane, blood bicarbonate profile and the total balance of the sessions were studied in six dialysis patients under the same operating conditions over 36 HDF sessions, in order to compare the effects of predilution HDF (pre-HDF), postdilution HDF (post-HDF), and mixed HDF on the final bicarbonate balance. RESULTS: The final HCO3- balance was more positive in post-HDF vs pre-HDF (142 +/- 36 vs 99 +/- 41 mmol/session, p<0.05), with a final blood HCO3- concentration of 26.6 +/- 1.0 vs 25.4 +/- 1.1 mmol/L, (p<0.05). Mixed HDF yielded intermediate results (balance: 119 +/- 42 mmol/session, final HCO3- 26.2 (1.2 mmol/L). These differences were seen to result from the increased HCO3- concentration of blood entering the filter in predilution, due to the infused HCO3-, enhancing convective loss and reducing the driving force for diffusive HCO3- gain. CONCLUSIONS: Bicarbonate concentration in dialysate-reinfusate is critical in order to obtain an adequate end of session HCO3- balance in on-line HDF. The predilution method produced the lowest cumulative net HCO3- gain between the three studied infusion modes. Our data suggest that, under the same operating conditions and excluding the effect of ultrafiltration, dialysate HCO3- should be increased by about 2 mmol/L in pre HDF, and 1 mmol/L in mixed HDF, to yield the same final balance as in post-HDF. PMID- 11905513 TI - Dimensions of central venous structures in humans measured in vivo using magnetic resonance imaging: implications for central-vein catheter dimensions. AB - The tip of a central vein catheter for hemodialysis should be located in the upper right atrium for the best performance. Hemodialysis catheters do have internal diameters unadjusted to the catheter length; however, the longer the catheter the slower the flow at the same pressure difference. On the other hand, the catheter diameter cannot be so large as to fill the vein too tightly as it predisposes to damage of the vein wall, thrombosis and stenosis. Therefore, the catheter length and diameter should be appropriate for the patient. For this purpose, the exact dimensions of the venous system in vivo should be known. In this study we correlated the anthropometric measurements and the dimensions of the large upper body veins in 31 adult volunteers. After deep inspiration, magnetic resonance imaging of the chest was performed in three planes; the positions of specific points in the three-dimensional coordinate system were measured, and the distance to adjacent points was calculated according to the analytic geometry formula. The total length from the catheter entry point to the right atrium was the sum of distances between the adjacent points. The lengths of the veins were correlated with the body anthropometric measurements (height, weight, body surface area, bi-acromion span, and height plus bi-acromion span). The best overall correlations of the lengths and diameters of the large upper body veins are with the body surface area. A table is included to guide the selection of the total catheter length and diameter in relation to the body surface area and insertion site. PMID- 11905514 TI - Patient knowledge and interest on dialysis efficiency: a survey. AB - BACKGROUND: Therapeutic compliance is fundamental on dialysis; however following a therapy requires a prior understanding of it. Aim of the study was to assess the need and interest for information on dialysis efficiency and to prepare a dedicated teaching tool. METHODS: 72 patients, on hemodialysis in two limited care satellite units, were given a questionnaire testing knowledge and interest on dialysis efficiency. In a subsequent second phase, following patients' suggestions, a cartoon book was prepared and opinions recorded. RESULTS: 63 patients' returned the questionnaire. 79.4% had basic knowledge on routine blood tests, 30.1% were aware of their specific meaning. All patients asked for further information, preferring books to other media. The book "Kt/V as cartoon" was distributed; 71.2% read it, 93% scored it as good-very good. In the Unit employing flexible dialysis schedules, 22/42 patients increased dialysis time. CONCLUSIONS: Despite insufficient knowledge on dialysis efficiency, patient interest is high. An educational program is feasible and may also give practical results, such as self-increase in dialysis time. PMID- 11905515 TI - A simplified cardiopulmonary bypass technique for animal experiments on implantable ventricular assist devices. AB - We have developed and report on a simplified cardiopulmonary bypass technique for experiments on implantable ventricular assist devices in calves. We used an electromechanical implantable ventricular assist device with a double cylindrical cam in three calves. Cannulas for the ventricular assist system were designed to be inserted between the left atrium and the descending aorta. We used the outflow cannula of the ventricular assist device, anastomosed to the descending aorta, as a temporary arterial return route for the cardiopulmonary bypass. A cannula for venous drainage was iserted into the right ventricle through the pulmonary artery. There were no problems related to the procedure and the cardiopulmonary bypass was succesful. In conclusion, this simplified cardiopulmonary bypass technique without neck incision in calves, as used in developmental work involving implantable ventricular assist devices, can be reliably performed. PMID- 11905516 TI - Combined effect of oxygen and ammonia on the kinetics of ammonia elimination and oxygen consumption of adherent rat liver cells. AB - Oxygen is essential for the survival of isolated liver cells and its concentration is known to affect their viability and function. Recent reports have also shown that ammonia is eliminated at a rate depending on its concentration and that high ammonia concentrations may be cytotoxic to rat liver cells. Nonetheless, little quantitative information on the effect of either metabolite on liver cell reaction kinetics is available although important to the design of bioreactors for bioartificial livers (BALs). In this investigation, we characterized the dependence of the rate of oxygen consumption (OCR), ammonia elimination (AER) and urea synthesis (USR) on ammonia concentration at physiological (i.e., 43 and 72 mmHg) and supra-physiological (i.e., 134 mmHg) dissolved oxygen tensions. To this purpose, isolated rat liver cells were cultured in adhesion on collagen in a continuous-flow bioreactor optimised for the kinetic characterisation of liver cell metabolic reactions. Rates of the investigated reactions generally increased with increasing ammonia concentrations. OCR and USR significantly increased with increasing dissolved oxygen tensions, particularly at high ammonia concentrations. The actual dissolved oxygen tension significantly influenced also OCR and USR dependence on ammonia concentration. The best-fit rate equations were used to show that, at the beginning of the treatment with a bioreactor packed with primary liver cells, high ammonia concentration in the blood may cause large hypoxic zones in the bioreactor as a result of its effect on OCR. This suggests that plasma (or blood) detoxification prior to entering the bioreactor might enhance BAL efficacy by preserving a large fraction of the available cell activity for longer times. PMID- 11905517 TI - Novel hybrid artificial liver using hepatocyte organoids. PMID- 11905518 TI - Adult stem cell technology--prospects for cell based therapy in regenerative medicine. PMID- 11905519 TI - Serological and molecular testing in hepatitis B and the dialysis patient. PMID- 11905520 TI - Changes in bone mineral density with age in men and women: a longitudinal study. AB - We performed a prospective study to evaluate the normal changes in bone mineral density (BMD) in the forearm, hip, spine and total body, and to study the agreement between changes in BMD estimated from cross-sectional data and the actual longitudinal changes. Six hundred and twenty subjects (398 women, 222 men; age 20-89 years) without diseases or medication known to affect bone metabolism undertook baseline evaluations, and 525 (336 women, 189 men) completed the study. BMD was measured twice 2 years apart by dual-energy X-ray absorptiometry. From cross-sectional evaluations the only premenopausal bone loss (<0.003 g/cm2/year) was found in the hip. In women after menopause and in men an age-related bone loss (0.002-0.006 g/cm2/year) was found at all sites. The data from the longitudinal evaluation showed a small bone loss in women before menopause at the hip and lumbar spine (<0.4%/year (<0.004 g/cm2/year)); this bone loss nearly tripled in the early postmenopausal years (<10 years since menopause), and thereafter decreased to the premenopausal rate for the hip, and to zero for the lumbar spine. The most pronounced bone loss after menopause occurred in the forearm (1.2%/year (0.006 g/ cm2/year)), and it remained constant throughout life. In men there was a small longitudinal bone loss in the hip throughout life, and a small bone loss in the distal forearm after the age of 50 years. In all groups, except for the early postmenopausal women, we found a small increase in total body BMD with age. When comparing the changes in BMD estimated from cross sectional data with the longitudinal changes, only the hip and forearm generally displayed agreement, whereas the changes in the total body and spine generally were incongruous. In conclusion, the hip and forearm appear to be the sites with the best agreement between the cross-sectional estimated and the longitudinal age related changes in BMD. PMID- 11905521 TI - Evaluation of the age-adjusted incidence of hip fractures between urban and rural areas: the difference is not related to the prevalence of institutions for the elderly. AB - As many as 40% of hip fractures occur in institutions for the elderly. Several studies have demonstrated a higher age-adjusted incidence of hip fractures in urban areas compared with rural areas. To assess whether this difference could be due to a preferential location of institutions for the elderly in urban areas, we compared the incidence of hip fractures over a 5-year period in urban versus rural areas, as defined according to the population density (urban > 15 inhabitants/ha2). We then determined the age-adjusted incidence of hip fractures in institutional-dwelling elderly and home-dwelling elderly. Hip fracture incidence was 100.0/100,000 (150.5 in women and 43.8 in men) in urban areas, and 71.0/100,000 (107.2 in women and 32.8 in men) in rural areas (p<0.001). When only those patients living in their own homes were analyzed, the incidence was 66.7/100000 (94.6 in women and 35.7 in men) in urban regions and 36.8/100,000 (49.6 in women and 23.4 in men) in rural areas (p<0.001), a difference of even greater magnitude than when both home-dwelling and institutional-dwelling residents were considered together. In a logistic regression model including age class, gender, urban or rural areas and institutionalization for inhabitants 65 years of age and older, urban residents have a 31% significantly (p<0.001) higher incidence of hip fracture compared with rural residents; women have a 79% significantly (p<0.001) higher incidence of hip fracture compared with men; and institutional-dwelling elderly have a 351% significantly (p<0.001) higher incidence of hip fracture compared with home-dwelling elderly. These results confirm the existence of a higher age-adjusted incidence of hip fractures in urban compared with rural areas. Since this difference is increased when patients living at home were analyzed separately, it indicates that the difference between urban and rural areas is not due to a preferential urban location of institutions for the elderly. PMID- 11905522 TI - Changes in calcaneal trabecular bone structure assessed with high-resolution MR imaging in patients with kidney transplantation. AB - The purpose of this study was to use high-resolution magnetic resonance (HR-MR) imaging to analyze the trabecular bone structure of the calcaneus in patients before and after renal transplantation and to compare this technique with bone mineral density (BMD) in predicting therapy-induced bone loss and osteoporotic fracture status. HR-MR imaging (voxel size: 0.195 x 0.195 x 1 mm) was performed at 1.5 T with an axial and sagittal orientation in 48 patients after transplantation, 12 patients before renal transplantation and 20 healthy controls. Structure measures analogous to standard histomorphometry and fractal dimension were determined in these images. BMD measurements of the lumbar spine and the proximal femur were obtained in the healthy female controls and the patients. Vertebral and peripheral fracture status were determined in all patients. The structural measures app.BV/TV, Tb.Sp, Tb.Th and Tb.N showed significant differences between controls and patients (p<0.05) while fractal dimension showed no significant differences. Neither the structural measures nor BMD showed significant differences between patients before and after transplantation. Correlations between time after transplantation versus structural measures and BMD were not significant. Differences between fracture and nonfracture patients were significant for the structural measures app.BV/TV, Tb.Sp and Tb.N (axial images) as well as for app.Tb.Th (sagittal images) and spine BMD (p<0.05) but not for hip BMD. Using odds ratios the strongest discriminators between patients with and without fractures were app. BV/TV, app.Tb.Sp (axial images) and app.Tb.Th (sagittal images), even after adjustment for age and BMD. Using receiver operating characteristic analysis the highest diagnostic performance was found for a combination of BMD and structural measures. In conclusion, our results indicate that structural measures obtained from HR-MR images may be used to characterize fracture incidence in kidney transplant patients; the best results, however, are obtained using a combination of BMD and structural measures. PMID- 11905523 TI - New model-independent measures of trabecular bone structure applied to in vivo high-resolution MR images. AB - Complementing measurements of bone mass with measurements of the architectural status of trabecular bone is expected to improve predictions of fracture risk in osteoporotic patients and improve the assessment of response to drug therapy. With high-resolution MRI the trabecular network can be imaged with 156 x 156 x 500 microm3 voxels, sufficient to depict individual trabeculae, albeit with inaccurate thickness. In this work, distance transformation techniques were applied to the three-dimensional image of the distal radius of postmenopausal patients. Structural indices such as trabecular number (app.Tb.N), thickness (app.Tb.Th) and separation (app.Tb.Sp) were determined without model assumptions. A new metric index, the apparent intra-individual distribution of separations (app.Tb.Sp.SD), is introduced. The reproducibility of the MR procedure and structure assessment was determined on volunteers, and the coefficient of variation was found to be 2.7-4.6% for the mean values of structural indices and 7.7% for app.Tb.Sp.SD. The distance transformation methods were then applied to two groups of patients: one of postmenopausal women without vertebral fracture and one of postmenopausal women with at least one vertebral fracture. It was found that app.Tb.Sp.SD discriminates fracture subjects from non-fracture patients as well as dual-energy X-ray absorptiometry (DXA) measurements of the radius and the spine, but not as well as DXA of the hip. Using receiver operating characteristic analysis, the area under the curve (AUC) values were 0.67 for app.Tb.Sp.SD, 0.72 for DXA radius, 0.67 for DXA spine and 0.81 for DXA of the hip. A combination of MR indices reached an AUC of 0.75. Age-adjusted odds ratio ranged from 1.85 to 2.03 for app.Tb.N, app.Tb.Sp and app.Tb.Sp.SD (p<0.003). We conclude that in vivo high-resolution MRI not only has the potential of imaging trabecular bone, but in combination with novel metrics may offer new insight into the structural changes occurring in postmenopausal women. PMID- 11905524 TI - Long-term follow-up of bone mass after orthotopic liver transplantation: effect of steroid withdrawal from the immunosuppressive regimen. AB - Glucocorticoids have been suggested to play a major role in transplantation related osteopenia. In this study we assess the long-term changes and the effect of steroid withdrawal from the standard immunosuppressive regimen on bone mineral density (BMD) after orthotopic liver transplantation (OLT). Sixty-nine non osteoporotic patients (20 women, 49 men), aged 48 +/- 9.5 years (mean +/- SD), and with a follow-up of 58.3 +/- 23.2 months (range 24-121 months) were studied. Immunosuppressive treatment consisted of prednisone, cyclosporin A and azathioprine. In 41 patients (group A), prednisone was tapered and withdrawn after 36.2 +/- 19.3 months (range 13-79 months), whereas in 28 patients (group B) prednisone was maintained. BMD in the spine (L1-L4) was serially measured by dual energy X-ray absorptiometry (Hologic QDR 1,000w) at baseline, before steroid withdrawal and at the end of study. Age- and sex-matched Z-scores of BMD were calculated. No differences were found in age, body mass index, time since OLT, or baseline BMD between the two groups. BMD had significantly increased in both groups at the end of follow-up period (group A, +8.1 +/- 8.7%; group B, +3.2 +/- 8.0%, p < 0.05). However, the Z-score was significantly higher in group A than in group B at the end of study (-0.44 +/- 1.05 vs -0.99 +/- 0.77; p<0.05). BMD recovery was lower in pre-OLT biliary cirrhosis patients. Bone mass improvement was independent of the time since OLT in both groups, and of the time of steroid withdrawal in group A. Our data confirm that steroid withdrawal accelerates the recovery of bone mass in patients who have undergone a successful liver transplantation. PMID- 11905525 TI - Abnormal bone turnover in cystic fibrosis adults. AB - Cystic fibrosis (CF) patients often have low bone mineral density (BMD) and may suffer from fractures and kyphosis. The pathogenesis of low BMD in CF is multifactorial. To study bone metabolism, we collected fasting serum and urine from 50 clinically stable CF adults (mean age 28 years) and 53 matched controls to measure markers of bone formation and bone resorption. The CF subjects had moderate lung disease (FEV1: 46.1 +/- 18.6% predicted) and malnutrition (BMI: 20.0 +/- 3.3 kg/m2). Only 3 subjects had normal BMD. CF subjects had higher urinary N-telopeptides of type I collagen (81.0 +/- 60.0 vs 49.0 +/- 24.2 nm BCE/mmol creatinine, p = 0.0006) and free deoxypyridinoline (7.3 +/- 5.0 vs 5.3 +/- 1.9 nM/mM, p = 0.004) levels than controls. Serum osteocalcin levels were similar in the two groups, a result confirmed by two immunoassays that recognize different epitopes on osteocalcin. Serum bone-specific alkaline phosphatase levels were elevated in CF patients (32.0 +/- 11.3 vs 21.8 +/- 7.0 U/l, p < 0.0001), but were much more closely associated with serum total alkaline phosphatase levels (r = 0.51, p = 0.001) than with age or gender. Parathyroid hormone levels were elevated (p = 0.007) and 25-hydroxyvitamin D levels were depressed (p = 0.0002) in the CF patients in comparison with controls. These results indicate that adults with CF have increased bone resorption with little change in bone formation. Medications that decrease bone resorption or improve calcium homeostasis may be effective therapies for CF bone disease. PMID- 11905526 TI - Gender differences in serum markers of bone resorption in healthy subjects and patients with disorders affecting bone. AB - To assess how two different serum markers of bone resorption may reflect changes in bone turnover, we compared age- and sex-related changes in serum C-terminal telopeptide of type I collagen (betaCTx) and tartrate-resistant acid phosphatase activity (TRAP) in 136 healthy men and 184 normal women. Serum levels of the two markers were also assessed in several groups of patients of both sexes presenting with the most common metabolic and endocrine bone diseases: established osteoporosis (n = 77), primary hyperparathyroidism (n = 44), glucocorticoid excess (n = 17), chronic renal failure (n = 39), active Paget's disease of bone (n = 5), humoral hypercalcemia of malignancy (n = 3), osteomalacia (n = 3), hyperthyroidism (n = 10), post-surgical hypoparathyroidism (n = 10), acromegaly (active disease, n = 8) and Cushing's syndrome (n = 10). In men the regression of betaCTx with age showed an initial decrease in bone resorption followed by an increase thereafter, starting from the sixth decade of life. No age-related change in serum TRAP activity was observed. In women, by contrast, a slight but significant linear correlation of both serum betaCTx and TRAP with age (r = 0.223, p<0.003 and r = 0.333, p<0.0001, respectively) was found, the two markers being positively correlated (r = 0.238, p<0.002). In each class of patients the mean Z-scores of betaCTx were significantly higher than those of TRAP activity. Moreover, compared with normal subjects, serum betaCTx seems to be characterized by a superior sensitivity relative to TRAP measurement, at least in the disorders studied. PMID- 11905527 TI - Biomechanics of bone: determinants of skeletal fragility and bone quality. AB - Bone fragility can be defined by biomechanical parameters, including ultimate force (a measure of strength), ultimate displacement (reciprocal of brittleness) and work to failure (energy absorption). Bone fragility is influenced by bone size, shape, architecture and tissue 'quality'. Many osteoporosis treatments build bone mass but also change tissue quality. Antiresorptive therapies, such as bisphosphonates, substantially reduce bone turnover, impairing microdamage repair and causing increased bone mineralization, which can increase the brittleness of bone. Anabolic therapies, such as parathyroid hormone (PTH-(1-84)) or teriparatide (PTH-( 1-34)), increase bone turnover and porosity, which offset some of the positive effects on bone strength. Osteoporosis therapies may also affect bone architecture by causing the redistribution of bone structure. Restructuring of bone during treatment may change bone fragility, even in the absence of drug effects on bone mineral density (BMD). This effect may explain why some drugs can affect fracture incidence disproportionately to changes in BMD. For instance, in a recent clinical trial, PTH-(1-34) therapy caused a dose related increase in spinal BMD without any dose-dependent effect on the observed decrease in spinal fracture incidence. This apparent disassociation between spinal BMD and bone fragility is probably due to effects of PTH-(1-34) on bone architecture within vertebral bodies. While it has been shown that BMD is highly heritable, bone mineral distribution and architecture are also under strong genetic influence. Recent findings suggest that different genes regulate trabecular and cortical structures within lumbar vertebrae, producing a wide range of bone architectural designs. These findings suggest that there is no single optimal bone architecture; instead many different architectural solutions produce adequate bone strength. PMID- 11905528 TI - Quantum chemical predictions of the vibrational spectra of polyatomic molecules. The uracil molecule and two derivatives. AB - This work describes the different scaling procedures used to correct the quantum chemical theoretical predictions of the IR and Raman vibrational wavenumbers. Examples of each case are shown, with special attention to the uracil molecule and some derivatives. The results obtained with different semiempirical and ab initio methods, and basis sets, are compared and discussed. A comprehensive compendium of the main scale factors and scaling equations available to obtain the scaled wavenumbers is also shown. PMID- 11905529 TI - Photoacoustic spectra of some biological metal chelates. AB - Photoacoustic spectra of Ni2+ in three new chelates containing azolo-2,4 pentanedione derivatives have been recorded and analysed in the visible region. The approximate symmetry of Ni2+ has been found to be octahedral. The d-d transitions in the red region show some similarity with those of nickel chloride and nickel acetate. In each case two charge-transfer bands appear in the violet region. PMID- 11905530 TI - Temperature and substituent effects on the dissociation constants of 5 azorhodanine derivatives. Semi-empirical quantum mechanical calculation. AB - Detailed analysis of the electronic structure and properties of some rhodanine derivatives (RDs) is presented. The aim of the present investigation is to pinpoint the electronic structural similarities and differences, among the series of the studied RDs that govern and determine their acidic, basic and co ordinative properties. The geometries of the studied rhodanine were fully optimized at the level of AMI semi-empirical method. Relative stabilities of the enol/keto isomers have been calculated. Proton affinities and proton detachment energies were computed for the series of rhodanine studied, at the level of AM1 method and compared with the potentiometrically-determined proton-ligand dissociation constants. Zero-point energy and electron correlations have been taken into consideration. pK(H) have been found to increase with increasing electron-donating nature of the substituents. The resulting linear Hammett plots of pK(H) versus the Hammett constant sigma values indicate the co-planarity of the investigated molecules. The evaluated thermodynamic parameters (deltaG, deltaH and deltaS) indicate that the dissociation processes are non-spontaneous, endothermic and entropically unfavourable. PMID- 11905531 TI - Study on the resonance light scattering spectrum of berberine cetyltrimethylammonium bromide system and the determination of nucleic acids at nanogram levels. AB - The interaction of berberine with nucleic acid in the presence of cetyltrimethylammonium bromide (CTMAB) in aqueous solution has been studied by spectrophotometry and resonance light scattering (RLS) spectroscopy. At pH 7.30, the RLS signals of berberine were greatly enhanced by nucleic acid in the region of 300-600 nm characterized by four peaks at 324.0, 386.5, 416.5 and 465.0 nm. The binding properties were examined by using a Scatchard plot based on the measurement of enhanced RLS data at 416.5 nm. Under optimum conditions, the increase of RLS intensity of this system at 416.5 nm is proportional to the concentration of nucleic acid. The linear range is 7.5 x 10(-9)-7.5 x 10(-5) g ml(-1) for calf thymus DNA, 7.5 x 10(-9)-2.5 x 10(-5) g ml(-1) for herring sperm DNA, and 5.0 x 10(-9)-2.5 x 10(-5) g ml(-1) for yeast RNA. The detection limits (S/N = 3) are 2.1 ng ml(-1) for calf thymus DNA, 6.5 ng ml(-1) for herring sperm DNA and 3.5 ng ml(-1) for yeast RNA, respectively. Three synthetic samples were analyzed satisfactorily. PMID- 11905532 TI - Conformations of dimethoxymethane: matrix isolation infrared and ab initio studies. AB - Conformations of dimethoxymethane (DMM) were studied using matrix isolation infrared spectroscopy. DMM was trapped in an argon matrix using an effusive source at 298, 388 and 430 K. Experiments were also done using a supersonic jet source to look for conformational cooling in the expansion process. As a result of these experiments, spectrally resolved infrared features of the ground and first higher energy conformer of DMM have been recorded, for the first time. The experimental studies were supported by ab initio computations performed at HF and B3LYP levels, using a 6-31++G** basis set. Computationally, four minima were identified corresponding to conformers with GG, TG, G+G- and TT structures. The computed frequencies at the B3LYP level were found to compare well with the experimental matrix isolation frequencies, leading to a definitive assignment of the infrared features of DMM, for the GG and TG conformers. At the B3LYP/6 31++G** level, the energy difference between the GG and TG conformers was computed to be 2.30 kcal mol(-1). The barrier for conformation interconversion, TG-->GG level was calculated to be 0.95 kcal mol(-1). This value is consistent with the experimental observation that the spectral features due to the TG conformer disappeared in the matrix on annealing. PMID- 11905533 TI - Infrared spectroscopy of goethite dehydroxylation. II. Effect of aluminium substitution on the behaviour of hydroxyl units. AB - Dehydroxylation of goethite as affected by aluminium substitution was investigated using Fourier transform infrared spectroscopy (FT-IR) in conjunction with X-ray diffraction (XRD), thermogravimetric analysis (TGA) and differential thermogravimetric analysis (DTGA). The band intensities of hydroxyl vibrations were indicative of the degree of dehydroxylation and the changes in band parameters due to aluminium substitution were observed. The effect of aluminium substitution on band parameters of FT-IR spectra of goethite and its partially and fully dehydroxylated products, the mixture of goethite/hematite and hematite, were interpreted. The results of this study have confirmed that aluminium substituted goethite is thermally more stable than non-substituted goethite and is in harmony with the results of XRD and DTGA. A larger amount of non stoichiometric hydroxyl units is associated with a higher aluminium substitution. A shift to a higher wavenumber of bending and hydroxyl stretching vibrations is attributed to the effects of aluminium substitution associated with non stoichiometric hydroxyl units on the a-b plane relative to the b-c plane of goethite. The results provide information for the characterisation of activated bauxite containing hematite and goethite. PMID- 11905534 TI - Spectral and photophysical studies of inclusion complexes of 2-amino-4,6-dimethyl pyrimidine with beta-cyclodextrin. AB - The interaction of 2-amino-4,6-dimethyl pyrimidine (ADMP) with beta-cyclodextrin (beta-CD) has been studied by means of UV absorption, steady state and time resolved fluorescence techniques. Spectral characteristics, bandwidths and photophysical parameters indicating that ADMP experience two different environments in aqueous solutions: bulk water and 1:1 (ADMP:beta-CD) inclusion complexation. The size restriction of the upper rim of beta-CD partially include ADMP and prevent the possibility of formation of 1:2 complex. The effective polarity of the cyclodextrin cavity experienced by the induced ADMP is equivalent with the polarity of an 80:20 methanol-water mixture. PMID- 11905535 TI - Resonance Rayleigh scattering technology as a new method for the determination of the inclusion constant of beta-cyclodextrin. AB - The interaction of procaine hydrochloride and beta-cyclodextrin in aqueous solution was studied using resonance Rayleigh scattering technology. The molar ratio of the inclusion complex was 1:1 established by spectrophotometry. The resonance Rayleigh scattering technology was first applied in the determination of the beta-cyclodextrin inclusion constant. The inclusion constant of procaine hydrochloride beta-cyclodextrin complex Kf is 1.23 x 10(2) and 1.27 x 10(2) l mol(-1) for method I and 1.15 x 10(2) and 1.21 x 10(2) l mol(-1) for method II. These determination results were in correspondence with the results of the spectrophotometric and fluorescence methods. Therefore, the resonance Rayleigh scattering method can be used as a new technology for the determination of the inclusion constant. PMID- 11905537 TI - On the interrelation between frequencies of stretching and bending vibrations in liquid water. AB - The problem is investigated whether the distributions of bending frequencies P(vbend,T) of H2O molecules in the liquid could be calculated from statistical distributions P(vOH,T) of stretching frequencies on the basis of the empirical correlation established for their mean values. It is found that correlations of different kinds fail to reproduce real spectra. They result in a bending band that is too narrow and give rise to an Evans hole in the stretching band. This provides evidence of the strong intermolecular coupling of bending modes, which makes their frequencies statistically independent from stretching frequencies. PMID- 11905538 TI - Quantitative analysis of (styrene/acrylonitrile/methyl methacrylate) co-polymer systems by infrared resonance spectroscopy. AB - A detailed careful analysis of the infrared resonance (IR) spectra of polystyrene (PSt), polymethyl methacrylate (PMMA), polyacrylonitrile (PAN) and their co mixtures were performed. Through this study the absorption peak area to weight ratios as well as working curves were obtained to test for their reliability as well as their suitability. Satisfactory results were achieved and these working curves were then used to measure the polymerized components of binary and ternary co-polymers. By investigating the acquired data we conclude that the monomer preferential polymeric sequence is St > MMA > AN. A quantitative method to measure P (St/AN/MMA) concentrations by IR spectroscopy is proposed in this work. PMID- 11905536 TI - Single crystal EPR studies on Mn(II)-doped sarcosine cadmium chloride and sarcosine cadmium bromide: study of zero-field splitting tensor in iso-structural complexes. AB - EPR spectra of single crystals of Mn(II)-doped sarcosine cadmium chloride and sarcosine cadmium bromide are studied in Q-band and in X-band at room temperature. Two magnetically inequivalent sites are observed in both the lattices in a distorted octahedral environment. The spin-Hamiltonian parameters are extracted and are found to have a rhombic symmetry. The angular variation of the zero-field transitions is simulated for one of the sites with an asymmetric zero-field tensor D = 480 x 10(-4) cm(-1), E = -115 x 10(-4) cm(-1) and a = 10 x 10(-4) cm(-1) for Mn(II) in sarcosine cadmium chloride and with D = 460 x 10(-4) cm(-1) E = -98 x 10(-4) cm(-1) and a = 10 x 10(-4) cm(-1) for Mn(II) in sarcosine cadmium bromide. The observed large value of zero-field tensor is due to the steric effects of the crystal packing caused by the ligands. Matumura's plot predicts an average covalency of 8.8 and 7.7% for the manganese-ligand bond in SCC and SCB lattices respectively. PMID- 11905539 TI - Studies of electron paramagnetic resonance parameters and defect structures for Ti2+ and V3+ ions in CdS crystals. AB - By applying the high-order perturbation formulas based on the cluster approach for the EPR parameters of 3d2 ions in trigonal symmetry, the zero-field splitting D, g factors gparallel, gperpendicular, and hyperfine structure constants Aparallel, Aperpendicular for Ti2+ and V3+ ions in CdS crystals are studied. From the studies, the defect structures of these paramagnetic impurity centers are obtained and the EPR parameters are also explained reasonably. PMID- 11905540 TI - A spectroscopic study of the fluorescence quenching interactions between biomedically important salts and the fluorescent probe merocyanine 540. AB - The effects of several biologically important inorganic salts, including NaCl, NaI, NaBr, KCl, MgCl2, MgSO4 and CaCl2 on the electronic absorption and fluorescence spectra of Merocyanine 540 (MC-540) have been investigated in aqueous media at 25 degrees C. Depending on both the MC-540 concentration and the nature of salt, a new absorption band appears at about 515 nm, above the critical salt concentration (CSC), corresponding to salt-induced MC-540 aggregation. Several types of MC-540 fluorescence quenching by the salts are observed, according to their cationic charge and the nature of anion: in the case of monovalent ions (Na+, K+), a non-linear Stern-Volmer behaviour is observed, indicating variable contributions of dynamic and static quenching mechanisms, whereas for divalent alkaline-earth (Mg2+, Ca2+) ions, linear Stern-Volmer relationships are obtained. Using these results, an analytical quenchofluorimetric approach is proposed for the determination of magnesium ions. PMID- 11905541 TI - Polarization IR spectra of hydrogen bonded pyrazole crystals: self-organization effects in proton and deuteron mixture systems. Long-range H/D isotopic effects. AB - This paper deals with experimental studies of the polarization IR spectra of solid-state pyrazole H1345, as well as of its H1D345, D1H345 and D1345 deuterium derivatives. Spectra were measured for the vN-H and vN-D band frequency ranges at temperatures of 298 and 77 K. The spectra were found to strongly change their intensity distribution and their polarization properties with the decrease of temperature. These effects were ascribed to some temperature-induced conformational changes in the hydrogen bond lattices. The studies reported allowed the finding of new kind of isotopic effects H/D in the open-chain hydrogen bond systems, i.e. the self-organization effects. It was found that the spectrally active aggregates of hydrogen bonds remain unchanged despite the growing isotope H/D exchange rate. This statement was supported by analysis of the residual polarized vN-H and vN-D band properties, measured for the isotopically diluted crystalline samples. Analysis of the band shapes of the four hydrogen isotope derivative crystals proved the existence of another kind of H/D isotopic effect, i.e. the long-range isotopic effect. It depends on an influence of the pyrazole ring hydrogen atoms onto the vN-H and vN-D band widths and onto the band fine structures. PMID- 11905542 TI - Spectroscopic studies on polymeric cobalt(II) and nickel(II) complexes with bridging succinonitrile and succinonitrile isotopomer ligands. AB - A series of cobalt(II) and nickel(II) complexes were synthesized using succinonitrile and its [1,4-13C2], [15N2]-, [2,2,3,3-2H4]- and [1,4-13C,-2,2,3,3 2H4]- isotopomers as bridging ligands. Spectroscopic studies, as well as X-ray powder diffraction profiles, were used to identify the nature of the octahedral coordination sphere of the central metal ions and to assign the vibrational spectra in full detail. The succinonitrile ligands were found to be in trans configuration in all the complexes studied and to be coordinated via the lone pairs of their nitrile nitrogens. The rule of mutual exclusion was found to be fulfilled for the succinonitrile ligands under the Ci symmetry of the complexes and the vibrations of the succinonitrile ligands were found to appear in either the infrared or the Raman spectra. All succinonitrile isotopomers exhibited blue shifts of 43-71 cm(-1) upon coordination, while most of the other vibrations remained unchanged or underwent small shifts of only a few wavenumbers. The mass differences of the succinonitrile isotopomers were found to shift mainly the vibrations of the respective affected part of the molecules in comparison with the normal succinonitrile. The exchange of the halides, which are coordinated to the central metal ion, was also found to influence the vibrations of the associated water molecules and we could identify vibrational bands arising due to the H-bond interaction between the halides and the water molecules. Finally, we showed that all complexes under consideration have, spectroscopically, the same symmetry. PMID- 11905543 TI - Analysis of UV-Vis spectral profiles of solvatochromic poly[3-(10-hydroxydecyl) 2,5-thienylene]. AB - The paper discusses a systematic study of the UV-Vis spectral profiles of poly[3 (10-hydroxydecyl)-2,5-thienylene] during its solvatochromic transformation in different solvent/non-solvent mixtures. A simulation of the overlapped spectra of the two chromophores (A less and B more conjugated) of the polymer is made through the resolution of their pure forms by means of vibronic progressions of log normal curves. The increment of the absolute intensity observed in the transformation has been determined and related to the increment of the transition moment; its value strongly supports the hypothesis that the solvatochromic transition consists in a doubling of A chromophores. Estimation of the average length of B chromophores has made it possible to evaluate the oscillator strength in the different solvent mixtures, the Franck-Condon factor and the Huang-Rhys parameter in relation to the intensity distribution of the vibronic structure. PMID- 11905544 TI - Fluorescence investigation of model compounds for light-emitting alternating copolymers in heterogeneous environments. AB - In this paper, the fluorescence spectra of the model compounds M(TPA-PPV) and M(TPA-PAV) of novel light-emitting alternating copolymers, were studied and the effect of KNO3 on the interaction between the model compounds and the ionic micelle-water interface was also investigated. The following is found: (i) the fluorescence changes of M(TPA-PPV) are related to the state of cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS) solution: (ii) an aggregated state can be formed in M(TPA-PAV) solution at low concentration of CTAB; (iii) higher concentrations of KNO3 may affect the interaction between the model compounds and the ionic micelle-water interface. PMID- 11905545 TI - Biological role of estrogen and estrogen receptors. PMID- 11905546 TI - DNA vaccine against malaria: a long way to go. AB - Vaccination is the attempt to mimic certain aspects of an infection for the purpose of causing an immune response that will protect the individual from that infection. Malaria, a disease responsible for immense human suffering, is caused by infection with Plasmodium spp. parasites, which have a very complex life cycle -antigenically unique stages infect different tissues of the body. It is a parasitic disease for which no successful vaccine has been developed so far, despite considerable efforts to develop a subunit vaccine that offers protective immunity. Due to the spread of drug-resistant malaria, efforts to develop an effective vaccine have become increasingly critical. DNA vaccination provides a stable and long-lived source of protein vaccine capable of inducing both antibody and cell-mediated immune responses to a wide variety of antigens. Injected DNA enters the cells of the host and makes the protein, which triggers the immune response. According to present needs, the flexibility of DNA vaccine technology permits the combination of multiple antigens from both the preerythrocytic and erythrocytic stages of malaria parasite. DNA vaccines with genes coding for different antigenic parts of malaria proteins have been created and presently some of these are undergoing field trials. The results from these trials will help to determine the likelihood of success of this technology in humans. This review presents an update of the studies carried out in malaria using DNA vaccine approach, the challenges, and the future prospects. PMID- 11905547 TI - Bronchodilating effect of oxitropium bromide in heart disease patients with exacerbations of COPD: double-blind, randomized, controlled study. AB - Anti-cholinergic agents are considered the bronchodilator therapy of first-choice in the treatment of patients with stable chronic obstructive pulmonary disease (COPD) associated with heart disease since they may be as effective or more effective than inhaled beta2-agonists and, moreover, they do not interact with cardiac beta-adrenoceptors. The aim of our study was to evaluate the bronchodilator activity of oxitropium bromide in outpatients suffering from exacerbations of COPD associated with heart diseases (ischaemic heart disease and/or arrhythmias). We recruited 50 consecutive outpatients (33 males and 17 females, mean age 68.6 years, 15 current smokers and 35 ex-smokers). Each patient performed body plethismography in basal condition and 30 min after inhalation of 200 microg metered dose inhaler (MDI) oxitropium bromide administered by a device (Fluspacer). FEV1, FVC, MMEF25-75, sRaw and tRaw were evaluated. Thirty minutes after 200 microg oxitropium bromide administration, we observed a significant improvement in FEV1 11.6% +/- 1 (mean +/- SEM) (P<0.01); FVC, MMEF25-75 sRaw variation was respectively: 9.2% +/- 0.6, 31.4 +/- 2.9, -19.9 +/- 1.1. Placebo did not significantly change pulmonary function. Our data suggest that oxitropium bromide bronchodilator activity is effective in exacerbations of COPD. PMID- 11905548 TI - Asthma control and differences in management practices across seven European countries. AB - Failure to follow asthma management guidelines may result in poor asthma control for many patients. The Asthma Insights and Reality in Europe (AIRE) survey, a multi-national survey assessing the level of asthma control from the patients perspective in seven Western European countries, previously demonstrated that the Global Initiative for Asthma (GINA) guideline goals were not achieved in Western Europe and that both adults and children with asthma were poorly controlled. Using additional data on asthma management practices from each of the seven countries in the AIRE survey, we compared variations in asthma morbidity and asthma management practices across countries to provide insight into the reasons for poor asthma control. Asthma management practices and asthma control among adults and children with current asthma were suboptimal in each of seven countries surveyed. Among patients with symptoms of severe persistent asthma, over 40% reported their asthma was well or completely controlled. School absence due to asthma was reported by upto 52.7% of children and up to 27.6% of adult reported work absence due to asthma. Lung function testing in the past year was uncommon: ranging from 13.5% of children in the U.K. to 68.8% of adults in Germany. Written asthma management plans were used by less than 50% of adults and less than 61% of children in all seven countries. Most adults (49.5-73.0%) and a large proportion of children (38.4-70.6%) had follow-up visits for their asthma only when problems developed. The ratio of recent inhaled corticosteroid use to recent short-acting beta-agonist use was inappropriate (<1) among patients with symptoms of severe asthma in all countries. This disparity was greatest among adults in Italy and France, where recent inhaled corticosteroid use was reported by less than one in nine patients reporting recent use of short-acting bronchodialators (IS:SAB <0.11). Management practices differ between countries and additional public health interventions and resources may be necessary to reduce patient suffering. Further efforts to fully implement asthma management guidelines are required to improve asthma control in Europe. PMID- 11905549 TI - Relationship between the perception of dyspnoea and airway inflammatory markers. AB - Poor dyspnoea perception in asthmatic patients seems to be associated with increased risk of asthma exacerbation. We have studied the relationship between basel ne dyspnoea perception and inflammatory markers in sputum in eight patients with mild asthma and in 13 patients with moderate to severe asthma. The perception of dyspnoea was scored on the Borg scale. Eosinophilic cationic protein (ECP) was measured by fluoroimmunoassay and by an interleukin (IL)-5 sandwich ELISA. The baseline Borg score was significantly higher in patients with severe asthma than in patients with mild to moderate asthma (4.1 +/- 0.29 vs. 2.28 +/- 0.28, P<0.05). The proportion of eosinophil and ECP levels in the sputum were significantly higher in patients with moderate to severe asthma. IL-5 in sputum was significantly increased in moderate to severe asthmatic patients compared to mild asthmatic patients. A significant relationship was found between the baseline perception score and FEV1/FVC (r = -0.53, P<0.01), sputum eosinophils (r = 0.70, P<0.01) and sputum ECP (r = 0.62, P<0.01). These findings suggest that the baseline perception score is related to inflammatory markers in sputum, and that the perception of dyspnoea as well as airway inflammatory markers may be considered to evaluate asthma severity. PMID- 11905550 TI - The prevalence of asthma and wheezing illnesses amongst 10-year-old schoolchildren. AB - Asthma and wheezing illnesses carry a significant burden of disease during childhood. Prevalence studies have the capacity to provide invaluable insights into the nature of these common conditions. As part of the Isle of Wight Whole Population Birth Cohort Study (n=1456) we have examined wheezing and asthma development amongst 10-year-old children. At this age 1373 children completed ISAAC written questionnaires whilst 1043 children performed further testing including skin-prick testing, serum inhalant IgE antibody screening, spirometry and bronchial challenge. At 10-years, prevalence of current wheeze was 18.9%, current asthma (symptomatic bronchial hyper-responsiveness--BHR) 14.4% and currently diagnosed asthma (current wheeze and asthma ever--CDA) 13.0%. Both wheezing and asthma at 10 years were associated with average symptom onset at 3 years of age indicating an early life origin for such conditions. Current wheeze (P=0.011) and CDA (P=0.008) showed significant male predominance. Considerable disease morbidity was identified for these states that tended to be greatest amongst children defined asthmatic rather than simply current wheezers. Wheezing and asthma were significantly associated with both atopy (P<0.001) and allergic co-morbidity Children with these states, particularly current asthma, also demonstrated impaired lung function (FEV1, P<0.001 and FEV1/FVC, P=0.010) and increased BHR (inverse slope, P<0.001). In conclusion, Asthma and wheezing showed substantial prevalence at 10 years of age. Strong associations to male gender, atopy, impaired lung function and BHR were seen for both wheeze and asthma. In regard to prevalence and morbidity characteristics, a questionnaire-based definition of currently diagnosed asthma gave similar results to the use of symptomatic BHR in defining current asthma. PMID- 11905551 TI - Assessment of asthma severity and treatment by GPs in Belgium: an Asthma Drug Utilization Research Study (ADUR). AB - A prospective study was performed between June 1996 and December 1997, to identify how general practitioners (GPs) in Belgium assess asthma severity and how they treat asthma according to their severity assessment. Three hundred and sixty-five GPs included 1376 already diagnosed and treated asthmatics. The GPs used a questionnaire providing data on patient demographics, aetiology of asthma, symptoms and medication use. The patients provided a complete diary card of day and night symptoms and morning and evening peak expiratory flow rates during a 3 week period. Asthma severity as assessed by GPs was compared with the severity according to the GINA guidelines. Along the same line, asthma treatment was evaluated according to the GPs assessment of severity and according to the GINA guidelines. Confronting the assessment of asthma severity by the GPs with the GINA criteria revealed that about 20% and 2% of the patients' asthma severity respectively were under- and over-estimated, respectively (using a discrepancy between GPs and GINA assessment of severity by 2 or more classes). Using the GINA criteria for treatment, only 37.5% of the patients seemed to be correctly treated. Taking a discrepancy between GINA assessment and treatment of two classes as an error, 2.3% and 23.4% of the asthmatics are over- and under treated, respectively. In conclusion, this study provides evidence that GINA guidelines seem not to be adequately interpreted and implemented by GPs in Belgium. Improvement of the assessment of asthma severity is definitely needed and may lead to more appropriate use of asthma medication. PMID- 11905552 TI - Lung abscess in adults: clinical comparison of immunocompromised to non immunocompromised patients. AB - Information related to the clinical characteristics and isolated microbes associated with lung abscesses comparing immunocompromised (IC) to non immunocompromised (non-IC) patients is limited. A retrospective review for 1984 1996 identified 34 consecutive adult cases of lung abscess (representing 0.2% of all cases of pneumonia), including 10 non-IC and 24 IC patients. Comparison of age, gender, tobacco use, pre-existing pulmonary disease or recognized aspiration risk factors were not significantly different between the two groups. Upper lobe involvement accounted for the majority of cases, although multi-lobe involvement was limited to IC patients. There were no differences in the need for surgical intervention, and mortality was very low for both groups. Anaerobes were the most frequent isolates for non-IC patients (30%), whereas aerobes were the most frequent isolate for IC patients (63%). Importantly, certain organisms were exclusively isolated in the IC group and multiple isolates were obtained only from the IC patients.Thus, comparing non-IC to IC patients, clinical characteristics may be similar whereas important differences may exist in the microbiology associated with lung abscess. These findings have important implications for the clinical management of these patient groups, and support a strategy to aggressively identify microbial agents in abscess material. PMID- 11905553 TI - Influence of deficient alpha1-anti-trypsin phenotypes on clinical characteristics and severity of asthma in adults. AB - Severe alpha1-anti-trypsin (AAT) deficiency implies a high risk of pulmonary emphysema development. The possible relationship between partial deficiencies of this enzyme and bronchial asthma remains controversial. The objective of this study was to ascertain the distribution of AAT phenotypes in a non-selected asthmatic patient population. Across-sectional study on a sample of 111 patients with asthma was carried out. Demographic and clinical variables were collected with serum IgE concentrations, plasma eosinophil number and serum AAT concentrations determined, together with the Pi phenotype. Asthma was mild in 36 (32.4%) patients, moderate in 45 (40.5%) and severe in 30 (27%). No differences were observed in eosinophil count or serum IgE or AAT concentrations among patients with different degrees of severity. Twenty-two (19.8%) asthmatics with deficient phenotypes for AAT were identified, distributed equally in all severity stages of the disease. No significant differences were found in clinical and functional characteristics, or in asthma morbidity between PiMM and PiMS patients or the heterozygote group (PiMS and PiMZ). Eosinophil count and IgE concentrations did not differ significantly between asthmatics with normal phenotype and heterozygotes. In conclusion, the distribution of AAT phenotypes in asthmatic patients did not differ from that found in the general population. Heterozygote phenotypes for the deficiency do not appear to confer greater severity or different clinical expression of asthma in adults. PMID- 11905554 TI - The clinical utility of arterialized earlobe capillary blood in the assessment of patients for long-term oxygen therapy (Respir Med 2001; 95: 655-660). PMID- 11905555 TI - Alpha1-anti-trypsin deficiency: epidemiology and incidence. PMID- 11905556 TI - The vibration of an artery-like tube conveying pulsatile fluid flow. AB - A hybrid method for investigating pulsatile fluid flow in a long, thin, artery like tube subjected to external excitations is presented. The non-linear partial differential equations governing the motion of the system, which incorporate the influence of circumferential strains, are solved by a combination of a finite element method, a finite difference method and a method of characteristics with interpolation. An initially axially stretched elastic tube conveying pulsating fluid, simply supported at both ends, is modelled to assess the effect of external harmonic excitation on the dynamic responses of the tube and the fluid flow. The results agree well with new experimental data. Comparison of the predicted results with those of a decoupled model demonstrates that it is necessary to consider the mechanism of fluid-structure interaction fully in the study of initially stretched cylindrical tubes conveying pulsatile fluid flow. An analysis of these coupling effects is presented for Womersley numbers alpha = 2.81 and 3.97 and a mean flow Reynolds number Re = 875. PMID- 11905557 TI - Medical image based, preformed titanium membranes for bone reconstructions: design study and first clinical. AB - The currently used intralesional or marginal surgical treatment of a bone tumour in the extremities shows some shortcomings in providing a restoration of the mechanical strength of the bone and the containment of the used filling materials. The use of a medical image based, preformed and custom-made titanium membrane screwed onto the periosteal side of the bone is introduced. This study looks in detail into the design process and the biomechanical evaluation of such a membrane. The buckling strength of the membrane, the strength at the perforation holes and the strength of the screw-bone fixation are tested experimentally. The two latter experiments are performed with different screw types. From the performed tests it appears that a titanium membrane without a wave pattern, of 0.3 mm thickness, fixed to the bone with seven trabecular bone screws (4 mm diameter and 28 mm length) is capable of carrying the anticipated mechanical loads on the reconstructed tibia. The medical image based design methodology and the first clinical application of such a preformed and custom made titanium membrane are reported and discussed. The feasibility of preformed titanium membranes for bone reconstruction in tumour surgery is demonstrated. PMID- 11905558 TI - Measurement of the bony anatomy of the humerus using magnetic resonance imaging. AB - This study was undertaken during the development of a new humeral intramedullary nail. To determine the overall size of the nail it was necessary to have the dimensions of the humerus and intramedullary canal. The dimensions of the bony anatomy of the humerus were obtained using an open magnetic resonance imager. The right arm of 20 volunteers was scanned and the length and the dimensions of both the intramedullary canal and the cortical bone were measured. The diameter of the canal was found to be 12.1+/-2.6 mm (mean +/- standard deviation) with the middle 50 per cent of measurements between 10 and 14 mm. The overall diameter of the bone was 19.3+/-2.3 mm, with the middle 50 per cent of measurements between 18 and 21 mm. This study has shown that magnetic resonance imaging (MRI) is an effective method of collecting dimensional data from any part of the skeleton for the development of medical devices. PMID- 11905560 TI - Arch-shaped versus flat arthrodesis of the ankle joint: strength measurements using synthetic cancellous bone. AB - The aim of this study was to see if preservation of the arch shape of the ankle at arthrodesis contributes to stability. The ankle joint was simulated by paired blocks of a synthetic material corresponding to rheumatoid cancellous bone with low stiffness and strength. Flat end constructs with and without subchondral bone were compared with arch-shape constructs with and without subchondral bone. The pairs were fixed with two screws simulating an arthrodesis. These constructs were then tested to failure in four-point bending and torque. In four-point bending the subchondral bone increased the strength, regardless of shape. Stiffness was higher in the arch-shaped specimens but was not influenced by the subchondral bone. In torque, both arch-shape and subchondral bone increase the strength. Stiffness was increased by arch-shape but not subchondral bone. The results imply that the arch-shape and subchondral bone should be preserved when performing an ankle arthrodesis, especially in weak rheumatoid bone. PMID- 11905559 TI - Development of a model for occipital fixation--validation of an analogue bone material. AB - Several implant systems may be used to fuse the skull to the upper cervical spine (occipitocervical fusion). Current biomechanical evaluation is restricted by the limitations of human cadaveric specimens. This paper describes the design and validation of a synthetic testing model of the occipital bone. Data from thickness measurement and pull-out strength testing of a series of human cadaveric skulls was used in the design of a high-density rigid polyurethane foam model. The synthetic occipital model demonstrated repeatable and consistent morphological and biomechanical properties. The model provides a standardized environment for evaluation of occipital implants. PMID- 11905561 TI - Selection of elastomeric materials for compliant-layered total hip arthroplasty. AB - A selection procedure has been developed to identify suitable commercial materials for use in compliant-layer artificial hip joints. Mechanical requirements, notably hardness and strength, as well as biocompatibility, constituted the specification for the compliant layer. Applying these constraints, candidate materials were identified in a broad range of polymeric material classes. Detailed sourcing and literature searching helped to identify materials appropriate to the application, with suitable mechanical and physical properties, as well as a history of successful clinical use. Some likely materials were identified but were prohibited from further consideration by limited commercial availability. Physical and mechanical characterization together with literature data were used to determine the relative ranking of the candidate materials and through a weighted materials property selection procedure the materials of choice were identified. The linear segmented aromatic polyurethanes, Tecothane 1085 and Estane 5714F1, emerged as the preferred materials. PMID- 11905562 TI - Analysis of mixed lubrication mechanism in metal-on-metal hip joint replacements. AB - A simple mixed lubrication model has been developed to predict the asperity contact and wear for the metal-on-metal bearing couple for total hip joint replacements. It has been shown that the femoral head radius has a large effect on the predicted asperity contact and wear depending on the lubrication regime. An increase in the femoral head radius can lead to an increase in wear under a predominantly boundary lubrication regime, but this trend can be reversed under a mixed lubrication regime towards fluid film lubrication. These observations are consistent with the recent experimental findings from hip simulator studies by Smith and co-workers. PMID- 11905563 TI - Helicopter-based lidar system for monitoring the upper ocean and terrain surface. AB - A compact helicopter-based lidar system is developed and tested under laboratory and field conditions. It is shown that the lidar can measure concentrations of chlorophyll a and dissolved organic matter at the surface of water bodies, detect fluorescence spectra of ground vegetation at a distance of up to 530 m, and determine the vertical profile of light-scattering particle concentration in the upper ocean. The possibilities of the lidar system are demonstrated by detection of polluted areas at the ocean surface, by online monitoring of three-dimensional distribution of light-scattering layers, and by recognition of plant types and physiological states. PMID- 11905564 TI - Validation of a novel ultraviolet lidar system with relative Raman-scattering cross sections determined from atmospheric measurements. AB - We have developed an ultraviolet lidar system in which the upwelled laser beam and the telescope field of view can be made to overlap at any specified location in space. We refer to this system as the Selected Overlap Lidar Experiment. We discuss validation of our system by calculating relative Raman-scattering cross sections (with respect to the nitrogen scattering cross section) for oxygen and water vapor using data collected during field operations of our lidar. Our relative cross sections are consistent with those obtained by other researchers making similar measurements in laboratory environments. PMID- 11905565 TI - Piffalls in atmospheric correction of ocean color imagery: how should aerosol optical properties be computed? AB - Current methods for the atmospheric correction of ocean-color imagery rely on the computation of optical properties of a mixture of chemically different aerosol particles through combination of the mixture with it into an effective, single particle component that has an average refractive index. However, a multi component approach in which each particle type independently grows and changes its refractive index with increasing humidity is more realistic. Computations based on Mie theory and radiative transfer are used to show that the two approaches result in top-of-the-atmosphere radiances that differ more than the water-leaving radiance. Thus, proper atmospheric correction requires a multicomponent approach for the computation of realistic aerosol optical properties. PMID- 11905566 TI - Open multipass absorption cell for in situ monitoring of stratospheric trace gas with telecommunication laser diodes. AB - A two-mirror multipass absorption cell that is operated open to the atmosphere from a stratospheric balloon to monitor in situ methane (in the 1.65-microm region) and water vapor (in the 1.39-microm region) with telecommunication laser diodes is described. A small Cassegrain-type telescope is used to couple the cell simultaneously with two near-infrared InGaAsP laser diodes by means of optical fibers. The 1-m cell provides an absorption path length of 56 m. The optical cell was carefully designed to be free of incidental fringing in the 10(-5) absorption range. It is used in combination with a dual-beam detector to obtain a detection limit of 10(-5) absorption units, a large dynamic range of the measurements of many orders of magnitude, and a precision error in the concentration determination of a few percents. The optical arrangement of the cell and its ability to be used to detect in situ trace gas in the stratosphere, in severe environmental conditions, are exposed. PMID- 11905567 TI - Grazing-incidence iridescence from a butterfly wing. AB - The Troides magellanus butterfly exhibits a specialized iridescence that is visible only when its hind wings are both illuminated and viewed at near-grazing incidence. The effect is due to the presence of a constrained bigrating structure in its wing scales that has been previously observed in only one other species of butterfly (Ancyluris meliboeus). However, whereas the Ancyluris presents wide angle flickering iridescence, the Troides butterfly uses pigmentary coloration at all but a narrow tailored range of angles, producing a characteristic effect. PMID- 11905568 TI - Hollow fibers for delivery of harmonic pulses of Q-switched Nd:YAG lasers. AB - Flexible hollow fibers for delivery of the second, third, and fourth harmonic pulses of Q-switched Nd:YAG lasers are introduced. For the doubled (532-nm) wavelength, we fabricated a hollow fiber with an internal metal and polymer film by using a silver-mirror plating and a liquid-phase-coating method. For tripled (355-nm) and quadrupled (266-nm) Nd:YAG in the ultraviolet region, we fabricated aluminum hollow fibers with or without an internal polymer layer by using the metal-organic chemical-vapor deposition method. Both types of fiber show high stability for the transmission of high-peak power laser pulses with low transmission losses. PMID- 11905569 TI - In situ combustion measurements of CO, H2O, and temperature with a 1.58-microm diode laser and two-tone frequency modulation. AB - An optical near-infrared process sensor for electric are furnace pollution control and energy efficiency is proposed. A near-IR tunable diode laser has performed simultaneous in situ measurements of CO (1,577.96 nm), H2O (1,577.8 and 1,578.1 nm), and temperature in the exhaust gas region above a laboratory burner fueled with methane and propane. The applicable range of conditions tested is representative of those found in a commercial electric arc furnace and includes temperatures from 1,250 to 1,750 K, CO concentrations from 0 to 10%, and H20 concentrations from 3 to 27%. Two-tone frequency modulation was used to increase the detection sensitivity. An analysis of the method's accuracy has been conducted with 209 calibration and 105 unique test burner setpoints. Based on the standard deviation of differences between optical predictions and independently measured values, the minimum accuracy of the technique has been estimated as 36 K for temperature, 0.5% for CO, and 3% for H2O for all 105 test data points. This accuracy is sufficient for electric arc furnace control. The sensor's ability to nonintrusively measure CO and temperature in real time will allow for improved process control in this application. PMID- 11905570 TI - Analysis of cavities for self-starting Kerr-lens mode-locked lasers. AB - We used the ABCD law to derive the transfer matrix of beam propagation in a Kerr lens mode-locked laser. On the basis of the transfer matrix and the formulas of the cavity stability G parameters that differ from the simple cavity g parameters, we give the expressions for Kerr-lens mode-locking (KLM) strength and some equations for cavity geometric parameters. The expressions and the equations can be used to construct a laser resonator to achieve self-starting oscillation of KLM. In addition we made a numerical simulation of the KLM strength at different positions in a mode-locked Ti:sapphire laser. The numerical results provide a good explanation of some of the physical phenomena observed in our experiments. PMID- 11905571 TI - Neodymium:YLF lasers at 1053 nm passively mode locked with a saturable Bragg reflector. AB - We present the results obtained from three different setups of stable, diode laser-pumped Nd:YLF lasers that operate in the TEM00 mode at 1053 nm and are passively mode locked by a saturable Bragg reflector. We obtained pulse widths tailored between approximately 70 ps and approximately 4 ns by using intracavity etalons of various thicknesses. With a maximum output power of 680 mW, we achieved an optical efficiency of 23%. PMID- 11905572 TI - Proton and gamma radiation tests on nonlinear crystals. AB - We report on the results of proton and gamma irradiation tests performed on nonlinear crystals for second- (SHG) and third-harmonic generation. Beta-barium borate (BBO), lithium triborate (LBO), and KTP crystals were exposed to three different energies of proton radiation (8, 70, and 300 MeV) and incremental doses of gamma radiation (up to 139 krad) in order to investigate the change in SHG performance and transmission spectra. BBO and LBO crystals turned out to be a suitable choice for SHG under radiative conditions. PMID- 11905573 TI - Phase-matched third-harmonic generation in mercury-(I)-chloride. AB - Using phase-matched third-harmonic generation we determine the effective nonlinear susceptibilities in Hg2Cl2 (Calomel) to /chi(3)eff,I/ = 4.5 x 10(-22) m2V(-2) and /chi(3)eff,II/ = 9.7 x 10(-22) m2V(-2) for type I and type II phase matching, respectively. The type III phase matching uses the same tensor components as type I and is deduced to be /chi(3)eff,III/ approximately equal to 1.5 x 10(-22) m2V(-2). The effective third-order susceptibilities of Hg2Cl2 are two orders of magnitude higher than those of CaCO3, and the tensor components chi11 - 3chi18 exceed the components of ADP by a factor of 5. These measurements demonstrate that Calomel might be a promising material to be used for nonlinear optical devices. PMID- 11905575 TI - Effect of terbium gallium garnet crystal orientation on the isolation ratio of a Faraday isolator at high average power. AB - We present a comprehensive and systematic investigation of the fundamental physical limitations of Faraday isolation performance at high average powers that are due to thermally induced birefringence. First, the operation of various Faraday isolator designs by use of arbitrary orientation of cubic magneto-optic crystals is studied theoretically. It is shown that, for different Faraday isolator designs, different crystal orientations can optimize the isolation ratio. Second, thermo-optic and photoelastic constants for terbium gallium garnet crystals grown by different manufacturers were measured. Measurements of self induced depolarization are made for various orientations of crystallographic axes. The measurements are in good agreement with our theoretical predictions. Based on our results, it is possible to select a crystal orientation that optimizes isolation performance at high average powers, resulting in a 5-dB enhancement over nonoptimized orientations. PMID- 11905574 TI - Optical parametric properties of ultraviolet-pumped cesium lithium borate crystals. AB - We report the optical parametric properties of cesium lithium borate (CLBO) crystals pumped by UV radiation of the fourth-harmonic generation at 266 nm and the third-harmonic generation at 355 nm of a picosecond Nd:YAG laser. A special optical design was used to avoid damage to the optical elements by the UV-pumped beam at 266 nm. The optical parametric generator (OPG)/optical parametric amplifier (OPA) of the 266-nm-pumped CLBO covers the tuning range from 347 nm in the UV to 1137 nm in the near IR. The 355-nm-pumped CLBO OPG/OPA, on the other hand, is tunable from 447 to 1725 nm. The experimental tuning curves for each CLBO OPG/OPA were measured and compared with the theoretical tuning curves. With a double-pass OPG configuration and a pumping intensity of approximately 6 GW/cm2, the maximum conversion efficiency, including both the signal and the idler, was approximately 11% for the 266-nm-pumped CLBO and is greater than 16% for the 355-nm-pumped CLBO without taking into account the surface losses from the uncoated elements. The bandwidth of this double-pass CLBO OPG at various wavelengths was measured and compared with other optical parametric systems. Because of the small angular dispersion of CLBO, the bandwidth of the OPG and OPA systems is exceptionally narrow, especially for the 266-nm-pumped system. Without the use of any dispersion element, the bandwidth of the 266-nm-pumped system can be as narrow as 0.22 nm at wavelengths far from the degenerate point. Comparison between the experimental bandwidth and the theoretical calculation shows that the bandwidth of the UV-pumped CLBO OPG/OPA is limited mainly by the divergence of the pump beam. PMID- 11905576 TI - Variational method for the retrieval of the optical thickness and the backscatter coefficient from multiangle lidar profiles. AB - A variational method for retrieving the aerosol optical thickness and backscatter coefficient profiles from multiangle lidar measurements is presented and discussed. A monostatic single-wavelength low-energy lidar system was operated at different zenith angles during the Indian Ocean Experiment (INDOEX) campaign in 1999 to characterize the aerosol plumes in the Indian monsoon. The variational method was applied to lidar data to retrieve profiles of optical thickness and the backscatter coefficient for nighttime and daytime measurements. Results are obtained with an uncertainty of 10% below 3 km (nighttime) and 2.8 km (daytime) and a bias of less than 0.01. During daytime the retrieval of optical parameters is indeed limited to a lower altitude owing to the sky background signal and the atmospheric inhomogeneity. In both cases the total aerosol optical thickness is consistent (+/- 10%) with the integrated value derived from sunphotometer measurements. Backscatter-to-extinction ratios estimated in different regions by two distinct methods compared well, which proves the capability of the method to assess optical measurements and account for the altitude dependence of the phase function. PMID- 11905577 TI - Fringe-imaging Mach-Zehnder interferometer as a spectral analyzer for molecular Doppler wind lidar. AB - The theoretical performance of a Mach-Zehnder interferometer used as a spectral analyzer for wind-speed measurement by direct-detection Doppler lidar is presented. The interferometer is optimized for the measurement of wind velocity from the signal that is backscattered by the molecules. In the proposed fringe imaging Mach-Zehnder (FIMZ) interferometer, a pattern of equally spaced linear fringes is formed and detected by two conventional detector linear arrays. Assuming a pure molecular signal with Gaussian spectral profile, an analytic expression for the standard deviation of the measurement error is obtained and compared with the Cramer-Rao lower bound given by an ideal spectral analyzer (ISA) in the case of shot-noise-limited signal. The FIMZ measurement error is shown to be 2.3 times that of the ISA and is comparable with the error given by previously developed multichannel spectral analyzers that are based on Fabry Perot interferometers that, in contrast, have the disadvantages of producing unequally spaced circular fringes and requiring dedicated detectors. The optimal path difference for a FIMZ operating at 355 nm is approximately 3 cm. The interferometer is shown to match important lidar beam etendues without significant performance reduction. PMID- 11905578 TI - Experimental determination of the lidar overlap profile with Raman lidar. AB - The range-dependent overlap between the laser beam and the receiver field of view of a lidar can be determined experimentally if a pure molecular backscatter signal is measured in addition to the usually observed elastic backscatter signal, which consists of a molecular component and a particle component. Two methods, the direct determination of the overlap profile and an iterative approach, are presented and applied to a lidar measurement. The measured overlap profile accounts for actual system alignment and for all system parameters that are not explicitly known, such as actual laser beam divergence and spatial intensity distribution of the laser light. PMID- 11905579 TI - Trace atmospheric SO2 measurement by multiwavelength curve-fitting and wavelength optimized dual differential absorption lidar. AB - We present a new differential absorption lidar (DIAL) method for atmospheric trace SO2 using multi-wavelength curve fitting. With this method we use five wavelengths around a SO2 absorption peak and obtain SO2 and O3 concentrations by fitting their absorption cross sections to measured DIAL and null results. A SO, concentration of 6 parts in 10(9) (ppb) was obtained for an altitude of 1050 m with 150-m range resolution. In addition, we optimized the wavelengths for dual DIAL SO2 measurement and demonstrated a high sensitivity of <0.5 ppb with 300-m range resolution. Comparison of these two methods is also presented. PMID- 11905580 TI - Heterodyne measurements of laser light scattering by a turbulent phase screen. AB - We report experiments in which a fiber-coupled heterodyne laser system operating at a wavelength of 1.5 microm is used to measure the phase fluctuations induced on a laser beam by passage through a thin layer of turbulent air and subsequent propagation through free space. We investigate the statistical properties and power spectra of the phase and its rate of change, in addition to the intensity statistics. We find that the power spectrum of the rate of change of phase has a simple negative exponential form. We discuss our results in the context of the problem of detection of phase variations over an extended turbulent atmospheric path. PMID- 11905581 TI - Ground-based full-sky imaging polarimetry of rapidly changing skies and its use for polarimetric cloud detection. AB - For elimination of the shortcomings of imaging polarimeters that take the necessary three pictures sequentially through linear-polarization filters, a three-lens, three-camera, full-sky imaging polarimeter was designed that takes the required pictures simultaneously. With this polarimeter, celestial polarization patterns can be measured even if rapid temporal changes occur in the sky: under cloudy sky conditions, or immediately after sunrise or prior to sunset. One of the possible applications of our polarimeter is the ground-based detection of clouds. With use of the additional information of the degree and the angle of polarization patterns of cloudy skies measured in the red (650 nm), green (550 nm), and blue (450 nm) spectral ranges, improved algorithms of radiometric cloud detection can be offered. We present a combined radiometric and polarimetric algorithm that performs the detection of clouds more efficiently and reliably as compared with an exclusively radiometric cloud-detection algorithm. The advantages and the limits of three-lens, three-camera, full-sky imaging polarimeters as well as the possibilities of improving our polarimetric cloud detection method are discussed briefly. PMID- 11905582 TI - Static Fourier-transform spectrometer with spherical reflectors. AB - A compact reflection Fourier-transform spectrometer without moving parts is developed. The spectrometer consists of two spherical reflectors: a Sagnac interferometer and a linear detector. The developed system is as small as 202 mm long x 185 mm wide x 100 mm high. The optics and the system configuration are described, and the preliminary experimental results are shown. PMID- 11905583 TI - Simultaneous vibrational and pure rotational coherent anti-stokes Raman spectroscopy for temperature and multispecies concentration measurements demonstrated in sooting flames. AB - The potential of measuring temperature and multiple species concentrations (N2, O2, CO) by use of combined vibrational coherent anti-Stokes Raman spectroscopy (CARS) and pure rotational CARS has been investigated. This was achieved with only one Nd:YAG laser and one dye laser together with a single spectrograph and CCD camera. From measurements in premixed sooting C2H4-air flames it was possible to evaluate temperatures from both vibrational CARS and rotational CARS spectra, O2 concentration from the rotational CARS spectra, and CO concentration from the vibrational CARS spectra. Quantitative results from premixed sooting C2H4-air flames are presented, and the uncertainties in the results as well as the possibility of extending the combined CARS technique for probing of additional species are discussed. PMID- 11905584 TI - Transportable automated ammonia sensor based on a pulsed thermoelectrically cooled quantum-cascade distributed feedback laser. AB - A compact ammonia sensor based on a 10-microm single-frequency, thermoelectrically cooled, pulsed quantum-cascade laser with an embedded distributed feedback structure has been developed. To measure NH3 concentrations, we scanned the laser over two absorption lines of its fundamental v2 band. A sensitivity of better than 0.3 parts per million was achieved with just a 1-m optical path length. The sensor is computer controlled and automated to monitor NH3 concentrations continuously for extended periods of time and to store data in the computer memory. PMID- 11905585 TI - Endothelial nitric oxide synthase gene Glu298Asp polymorphism and blood pressure, left ventricular mass and carotid artery atherosclerosis in a population-based cohort. AB - OBJECTIVE: Decreased production of endothelial nitric oxide (NO) is associated with different cardiovascular pathology. We studied the association between the Glu298Asp polymorphism of the NO producing gene, endothelial nitric oxide synthase (eNOS), and hypertension, left ventricular mass (LVM) and carotid artery intima-media thickness (IMT) in a population-based cohort of hypertensive and control subjects. DESIGN: Cross-sectional case-control study. SETTING: District around Oulu University Hospital, Northern Finland. SUBJECTS AND METHODS: The study population consisted of 600 middle-aged hypertensive subjects (300 men and 300 women) and 600 controls (300 men and 300 women) living in the City of Oulu. The hypertensive subjects were randomly selected by age stratification from the Social Insurance Institute register for reimbursement of antihypertensive medication. For each hypertensive subject, an age- and sex-matched control was randomly selected from the national health register. The overall participation rate was 87.8%. In the present study a total of 1024 subjects were screened. Echocardiographic examinations were performed by a trained cardiologist and carotid ultrasonographic examinations by a trained radiologist. RESULTS: The genotype distributions and allele frequencies between the hypertensive and control subjects and the relationship between the Glu298Asp variant and blood pressure, LVM and carotid artery IMT were determined. No differences in genotype distribution or allele frequencies were found between the hypertensive and control groups (the frequency of the Asp allele 0.299 vs. 0.288, respectively). Also, we could not find any association between the eNOS genotype and the measured cardiovascular complications. CONCLUSIONS: The Glu298Asp variant of the eNOS gene does not seem to be a major risk factor for cardiovascular alterations in the general population. PMID- 11905586 TI - Serum lipids, lipoprotein(a) level, and apolipoprotein(a) isoforms as prognostic markers in patients with coronary heart disease. AB - OBJECTIVE: Our objective was to study prognostic factors for death in patients with coronary heart disease (CHD), focusing on serum lipids and lipoproteins. DESIGN AND SUBJECTS: The study subjects were 964 patients with angina pectoris who underwent coronary angiography between 1985 and 1987. Follow-up, including survival and cause of death, was carried out in April 1998. RESULTS: A total of 363 patients died. Increasing age, diabetes and low levels of HDL cholesterol and of apolipoprotein (apo) AI were associated with increased risk of total mortality and cardiac mortality. In men, low levels of LDL cholesterol and of apoB were associated with increased risk of death, but not of cardiac death only; high levels of lipoprotein(a) [Lp(a)] were not associated with increased risk. In women, however, there was a trend towards increased risk with increasing Lp(a) levels (P = 0.054); the smallest isoform of apo(a) was associated with a twofold increase in risk. In women, but not in men, risk decreased with increasing molecular weight of the apo(a) isoforms. CONCLUSIONS: Amongst lipoprotein variables, low levels of HDL cholesterol and of apoAI and the presence of low molecular weight isoforms of apo(a) are associated with increased risk of death in patients with CHD. Apo(a) isoforms and Lp(a) levels seem to be more important as risk factors amongst women. Low LDL cholesterol and apoB levels were associated with increased risk, but only in men. These findings demonstrate the importance of a gender-specific analysis of risk factors for CHD. PMID- 11905587 TI - Detection and prevention of hepatitis C in dialysis patients and renal transplant recipients. A long-term follow up (1989-January 1997). AB - BACKGROUND: Hepatitis C is frequent problem in dialysis wards. DESIGN: A long time (1989-97) follow up of hepatitis C virus (HCV) infection in a Swedish nephrology unit was performed with anti-HCV screening, confirmatory antibody tests, viral RNA detection and molecular characterization. Case histories were reviewed with focus, onset of infection, liver morbidity and mortality. RESULTS: In October 1991, 10% (19 of 184) of the patients in the unit (haemodialysis-, peritoneal dialysis and transplanted patients) were verified or suspected HCV carriers, whilst the number at the end of 1996 was 8%, (13 of 157). Most patients were infected before 1991 but only in one case from a known HCV-infected blood donor. No new HCV infections associated with haemodialysis occurred during the study period. A total of 13 of 24 viremic patients had HCV genotype 2b, a pattern suggesting nosocomial transmission. This was further supported by phylogenetic analysis of HCV viral isolates in seven. HCV viremia was also common in patients with an incomplete anti-HCV antibody pattern as 8 of the 12 indeterminant sera were HCV-RNA positive. CONCLUSIONS: Awareness, prevention, identification of infected patients and donor testing limited transmission. Indeterminant recombinant immunoblot assays (RIBA)-results should be regarded with caution as a result of the relative immunodeficiency in uremic patients. Our data indicate nosocomial transmission in several patients. PMID- 11905588 TI - Relationship between anti-neutrophil cytoplasmic antibody determined with conventional binding and the capture assay, and long-term clinical course in vasculitis. AB - OBJECTIVES: To evaluate the relationship between anti-neutrophil cytoplasmic antibody (ANCA) measured with two different methods and long-term clinical course in vasculitis. DESIGN: Retrospective determination of ANCA with two different assays for detection of PR3-ANCA, conventional direct binding ELISA and capture ELISA using monoclonal antibodies against PR3. The 245 ANCA determinations were performed from frozen blood samples collected three to four times a year in each patient. SETTING: Department of Nephrology at a Swedish University Hospital. SUBJECTS: A total of 10 ANCA-positive patients with vasculitis caused by Wegener's granulomatosis (WG) or microscopic polyarteritis (MPA) and a very long follow-up time (mean 9 years, range 5-15.5 years). RESULTS: The total number of episodes with active vasculitis was 29 and all of them (100%) were detected by the capture technique whilst the conventional technique detected 23 (79%). The mean number of episodes with active disease requiring treatment with steroids and cytotoxic drugs was three per patient (range 1-6). At the time of clinical relapse of the vasculitis disease, the ANCA titre using the capture technique was either increasing or showed a very high value in all cases. The pattern of capture ANCA response could be subdivided into three categories: a close (four patients), an intermediate (three patients), and no (three patients) relationship between capture ANCA level and long-term clinical course. CONCLUSION: Detection of PR3-ANCA by the capture ELISA showed a higher sensitivity than that obtained by the direct ELISA in diagnosing relapse during follow-up of patients with vasculitis. The specificity of the capture ANCA was, however, low, as high levels occurred in patients without clinical disease activity. PMID- 11905589 TI - Reduction of free radical activity in amyloid deposits following liver transplantation for familial amyloidotic polyneuropathy. AB - OBJECTIVES: Liver transplantation halt the progress of familial amyloidotic polyneuropathy (FAP). Oxidative stress has been implicated in amyloid toxicity and formation. The objective of this study was to establish whether markers for oxidant stress and antioxidant capacity change following liver transplantation in patients with FAP. DESIGN: Morphometric and biochemical study. SETTING: Tertiary referral centre. SUBJECTS: Duodenal biopsy samples from 16 patients, taken before and after liver transplantation were used for morphometry. Serum samples from 14 patients, seven of whom had received transplants, were analysed regarding antioxidant capacity. INTERVENTION: Liver transplantation. MAIN OUTCOME MEASURES: Immunohistochemistry was used to stain for the lipid peroxidation product 4 hydroxynonenal (HNE), and Congo red staining was used for amyloid detection. Positive areas were quantified by point counting. Total antioxidant capacity (TAC) was measured with a colourimetric assay. RESULTS: In tissue, a decrease of HNE was noted after liver transplantation, whereas no significant changes were detected for amyloid deposits. No difference between transplanted and not transplanted patients was noted for total antioxidant capacity measured in serum. CONCLUSION: To our knowledge, this is the first description of a reduction of markers for free radical activity after cessation of amyloid formation. The findings implicate that amyloid formation in transthyretin (TTR) amyloidosis generates oxidative stress, whereas amyloid deposits as such are less toxic to sourrounding tissues. PMID- 11905590 TI - Chlamydia pneumoniae IgA- and IgG antibodies in young survivors of myocardial infarction. A comparison of antibody detection by a microimmunofluorescence test and an enzyme immunoassay. AB - OBJECTIVES: Chronic Chlamydia pneumoniae infection is considered as a cardiovascular risk factor and antibodies are commonly analysed by the subjective microimmunofluorescence (MIF) test. We wanted to investigate the C. pneumoniae IgA- and IgG seroprevalence in young survivors of myocardial infarction and matched controls, and to compare the agreement of detecting antibodies between a MIF test and an enzyme immunoassay (EIA). DESIGN: A total of 61 patients hospitalized as a result of myocardial infarction, 51 patients hospitalized with chest pain and negative exercise-ECG and 61 age and sex matched controls (mean age 53.3 years, range 40-60 years) were included in this case-control study. Serological comparisons were expressed as sensitivity, specificity and interrater agreement (K or Kw) of the EIA test related to the MIF test. RESULTS: Presence of IgA (cut off = 16) antibodies was significantly higher in coronary heart patients compared with controls for both assays (P = 0.02 by the MIF and P = 0.05 by the EIA test). The presence of IgG (cut off = 32) antibodies was significantly higher amongst patients (P = 0.05) when analysed by the MIF-test, but not with the EIA test (P = 0.16). The strength of agreement between the assays was good for both IgA- (Kw = 0.67) and IgG (Kw = 0.79) analyses. However, only 52.8% of the IgA samples classified as strong positive (cut-off = 32) by the MIF test were strong positive by the EIA test (K = 0.56). Only 73.2% of the negative IgG samples (<32) by the MIF-test turned out negative by the EIA-test (K = 0.73). CONCLUSIONS: Dependent on assay and cut-off level, there is an increased C. pneumoniae IgA- and IgG seroprevalence in young survivors of myocardial infarction compared with controls. Despite the subjective interpretation of MIF-titres, the strength of agreement between the EIA and MIF tests was good for both antibody classes. However, misclassification of highly positive IgA samples and negative IgG samples by the MIF test may influence study conclusions. We conclude that the choice of serological method is of major importance when evaluating a possible relationship between C. pneumoniae and coronary heart disease. PMID- 11905591 TI - Attitudes towards clinical research amongst participants and nonparticipants. AB - OBJECTIVES: To investigate attitudes to clinical research amongst cancer trial participants and nonparticipants, and to compare results with those from previous studies amongst participants in noncancer trials. DESIGN: Trial participating respondents were given three questionnaires during the clinical trials. Respondents amongst patients declining randomization answered a single questionnaire. SETTING: Participants and nonparticipants in randomized clinical cancer trials. SUBJECTS: Forty-one participants and 47 nonparticipants in cancer trials. RESULTS: Altruistic motives of physicians to conduct medical research were highly rated. Attitudes towards clinical research were positive in all groups, with nonparticipant respondents being the least positive. Eight to nine tenths found scientific testing necessary before general health service implementation. Trial participants were, as compared with nonparticipating respondents, more positive towards both participation of self and others. Both personal and altruistic motives for participation were highly rated. Primary reasons for nonparticipation were fear of 'the unknown' and/or unease with randomization. Only a minority felt a moral problem created by declining trial participation. Respondents amongst noncancer participants were more satisfied with the information given than both cancer participants and cancer nonparticipants. Negative experiences in cancer participants generally dealt with frustration related to seeing too many physicians at check-up appointments. CONCLUSION: Attitudes towards clinical research are generally positive even in cancer nonparticipants. Both personal and altruistic motives for participation were highly rated. A fear of 'the unknown' and resentments towards randomization were primary reasons to renounce participation. Seeing too many physicians at check-up appointments seems to be an important factor for negative experiences in cancer trial participants. PMID- 11905592 TI - High-dose intravenous immunoglobulin in the treatment of acute myocarditis. A case report and review of the literature. AB - A few reports have suggested the beneficial effect of high-dose intravenous immunoglobulin (IVIG) in the treatment of acute myocarditis and cardiomyopathy. We describe a 49-year-old woman in which acute myocarditis was diagnosed on the basis of clinical and echocardiographic findings. Conventional treatment with captopril and frusemide was administered: intravenous heparin and, subsequently, oral anticoagulants were added because of the appearance of an apical thrombus. On the fifth day of hospitalization, treatment with high-dose (400 mg kg(-1) day( 1)) IVIG was started and prosecuted for 5 days. A dramatic improvement of clinical conditions was observed, with increase of the left ventricular ejection fraction (LVEF) from 30% to 75% within 1 week. One year after the diagnosis the patient is in good health, with steady normal LVEF. The rapid recovery, which was immediately subsequent to the administration of high-dose IVIG, suggests that this kind of treatment has been effective in our patient with acute myocarditis. PMID- 11905593 TI - Risperidone and severe cerivastatin-induced rhabdomyolysis. PMID- 11905594 TI - Twenty-four hour blood pressure following stroke. PMID- 11905595 TI - Diabetes and advanced glycation endproducts. AB - Bio-reactive advanced glycation endproducts (AGE) alter the structure and function of molecules in biological systems and increase oxidative stress. These adverse effects of both exogenous and endogenously derived AGE have been implicated in the pathogenesis of diabetic complications and changes associated with ageing including atherosclerosis, renal, eye and neurological disease. Specific AGE receptors and nonreceptor mechanisms contribute to these processes but also assist in the removal and degradation of AGE. The final disposal of AGE depends on renal clearance. Promising pharmacologic strategies to prevent AGE formation, reduce AGE toxicity, and/or inactivate AGE are under investigation. PMID- 11905596 TI - Is red tape stifling the progress of new anticancer drugs? PMID- 11905597 TI - Alternative medicine and cancer patients in less developed countries. PMID- 11905598 TI - Crossed wires. PMID- 11905599 TI - Persistent human papillomavirus infection and cervical neoplasia. AB - The development of cervical cancer is preceded by precursor lesions (cervical intraepithelial neoplasia). Evidence-based epidemiological and molecular data suggest that persistent infections with human papillomavirus (HPV) types that carry ahigh oncogenic risk are the intermediate endpoints, leading to both intraepithelial and invasive cervical neoplasia. Integration of highly oncogenic HPVs into host-cell chromosomes is followed by binding of HPV E6 and E7 oncoproteins to tumour-suppressor genes p53 and RB, respectively. This process results in impaired tumour-suppressor-gene function, involving DNA repair, decreased apoptosis, and eventual cell immortalisation. Mutations causing chromosomal alterations, loss of heterozygosity, and proto-oncogene and telomerase activation in immunopermissive individuals have important roles in virus-induced cervical carcinogenesis. The so-called non-European variants of HPV 16 and 18 may increase the degradation potential of p53. HPV 16 is polymorphic and, although the evidence is controversial, the Arg/Arg genotype of p53 could have greater susceptibility to HPV-E6 degradation than the other genotypes. The coincident interplay between the non-European genomic variants of HPV 16/18 and p53 Arg/Arg may explain, at least in part, the persistence of HPV infection and tumour progression in women with cervical neoplasia. Further epidemiological and molecular research is needed, to gain insight into HPV-mediated cervical carcinogenesis. The evidence highlights the need to develop appropriate prophylactic HPV vaccines and diagnostic and screening tests. PMID- 11905600 TI - Oncolytic biotherapy: a novel therapeutic plafform. AB - There is a clear need for new, selective, cancer treatments that do not cause the cross-resistance which occurs with currently available chemotherapeutic agents. Gene therapy is a promising approach, but to date, it has shown limited effectiveness in clinical trials because of insufficient gene transduction. Many investigators are now revisiting the 'old' idea of using tumour-specific, replication-selective viruses or bacteria to treat cancer. These agents can be directly oncolytic, but can also be used to simultaneously express therapeutic genes in target cells or induce tumour-specific, cell-mediated immunity. We discuss the promise of this rapidly evolving field and examine the potential barriers to its success. PMID- 11905601 TI - Low susceptibility genes for breast cancer: the way forward? PMID- 11905602 TI - Progress report on the US war against cancer? PMID- 11905603 TI - Melanoma-stroma interactions: structural and functional aspects. AB - Cutaneous melanomas are notorious for their tendency to metastasise. Because the tumour microenvironment plays an important part in tumour development and progression, we review the structural and functional aspects of interactions between melanoma and the stroma. We emphasise fibrovascular patterns (both in uveal and cutaneous melanoma), cellular and extracellular composition of the stroma, and the molecules involved. Also, we discuss functional interactions, focusing on melanoma-fibroblast cross-talk by soluble factors and by direct cell cell contact. On the basis of recent findings we propose that involvement of fibroblasts in melanoma-stromagenesis occurs through different stages: recruitment, activation, and conversion to myofibroblasts, or differentiation to fibrocytes. We reason that this involvement is topographically linked to different areas in and around the tumour, and hypothesise that stromal activation, as seen in tumor ulceration or immunological regression in melanoma, stimulates tumour progression. PMID- 11905604 TI - Pain in Spain could be on the wane. PMID- 11905605 TI - Lymphatic dissemination of tumour cells and the formation of micrometastases. AB - Most human cancers show evidence of metastatic spread to regional lymph nodes, and the extent of lymph-node involvement is directly related to dinical outcome. Increased expression of vascular endothelial growth factor C in primary tumours is associated with increased dissemination of tumour cells to regional lymph nodes in various human carcinomas. Arguments favouring the activation of pre existing lymphatic endothelium and the de novo formation of lymphatic capillaries (lymphangiogenesis) are discussed. We highlight recent advances in the molecular detection and characterisation of lymph-node micrometastases, as well as potential microenvironmental factors, such as chemokines, which may influence the migration and growth of metastatic tumour cells. Finally, we examine the clinical significance of lymphatic-mediated tumour-cell dissemination and the formation of lymph-node micrometastases. PMID- 11905607 TI - Allan van Oosterom, President of the EORTC. European Organisation for the Research and Treatment of Cancer. Interview by Jeroen van der Boon. PMID- 11905606 TI - The seed and soil hypothesis: vascularisation and brain metastases. AB - The development of a relevant mouse model for the establishment and growth of brain metastases is essential for study of the biology and therapy of brain metastasis. Injection of human tumour cells into the internal carotid artery of syngeneic or nude mice produces experimental metastases in specific regions of the brain; these are not due to patterns of initial cell arrest, motility, or invasiveness, but rather to the ability of metastatic tumour cells to grow. Whether the progressive growth of brain metastases depends on neovascularisation is not clear. Immunohistochemical and morphometric analyses show that the density of blood vessels within experimental metastases in the brains of nude mice, or within brain metastases derived from human lung cancer, is lower than in the adjacent, tumour-free brain parenchyma. However, blood vessels associated with brain metastases are dilated and contain many dividing endothelial cells. Immunohistochemical analysis also reveals that tumour cells located less than 100 microm from a blood vessel are viable, whereas more distant tumour cells undergo apoptosis. The blood-brain barrier is intact in and around experimental brain metastases smaller than 0.25 mm in diameter, but is leaky in larger metastases. Nevertheless, the lesions are resistant to chemotherapeutic drugs. The way in which the brain microenvironment influences the biological behaviour of tumour cells is a subject of intense investigation. PMID- 11905608 TI - FDA licences imatinib mesylate for CML. PMID- 11905609 TI - Australian cancer survival figures greatly improved. PMID- 11905610 TI - The chicken or the egg? PMID- 11905611 TI - Spindle cell carcinomas are uncommon. PMID- 11905612 TI - Clinical testing of Ukrain. PMID- 11905613 TI - Importance of local tumour control in breast cancer. AB - The overall importance of local tumour control in the management of breast cancer, specifically the influence of local control on survival, remains one of the fundamental questions for oncologists. This review addresses the issues surrounding local tumour control, including the evolution of the concept of disease spread, the rationale for local control, the results of studies of radiotherapy after breast-conserving surgery and after mastectomy, and an interpretation of the recent data on post-mastectomy radiotherapy. PMID- 11905614 TI - Farnesyl transferase inhibitors: a novel targeted tnerapy for cancer. AB - Activating oncogenic mutations of the RAS gene are common in cancer, occurring in 30% of solid tumours in adults. Inhibitors of the enzyme farnesyl protein transferase prevent a key step in the post-translational processing of the RAS protein, and were developed initially as a therapeutic strategy to inhibit cell signalling in RAS-transformed cells. As more has been learnt about the biological effects of farnesyl transferase inhibitors on cancer cells, it has become increasingly clear that tumours without oncogenic RAS mutations may also be targets for farnesyl transferase inhibitor therapy. Encouraging results from phase I and II clinical trials have emerged, creating both enthusiasm and new challenges for the optimum clinical development of this important new class of anticancer drug. PMID- 11905615 TI - Cervical-cancer screening beyond the year 2000. AB - Evidence-based studies have shown that new techniques for cervical cancer screening have a higher diagnostic yield than conventional cervical cytology (Pap test). Automated screening devices that use liquid-based, thin-layer cytology and human papillomavirus DNA testing are likely to become the standard for routine primary screening for cervical cancer and its precursors in the 21st century. The increased initial expense of the new techniques will most certainly be absorbed by instituting longer intervals for safe primary screening, in both low-risk and high-risk populations. To make modern screening programmes even more effective, we must promote extensive public awareness campaigns about cervical cancer, a preventable disease. PMID- 11905616 TI - Apoptosis: implications of basic research for clinical oncology. AB - Recent studies have indicated a role for apoptosis in a variety of human diseases. Suppression of apoptosis contributes to carcinogenesis by several mechanisms, including facilitating the accumulation of gene mutations, permitting growth-factor-independent cell survival, promoting resistance to immune-based cytotoxicity, and allowing bypassing of cell-cycle checkpoints, which would normally induce apoptosis. Defects in apoptotic mechanisms also play an important part in resistance to chemotherapy and radiation. The core machinery of the cell death pathway can be reduced to a few critical types of proteins, which are well conserved across animal evolution. This review gives an update on the key players involved in apoptosis as well as an overview of the involvement of apoptosis in disease, and novel diagnostic and therapeutic options derived from the extensive basic research on this topic carried out over the last decade. PMID- 11905617 TI - Tobacco--on the side of the angels for once. PMID- 11905618 TI - Malignant melanoma presenting as nephrotic syndrome. PMID- 11905619 TI - The adolescent with cancer--at the edge of no-man's land. AB - The cancer journey of the adolescent patient can be difficult and lonely. Provision of services to these patients should occur at the interface between paediatric and adult oncology, although for the individual, that interface can seem like a gap into which he or she may too readily fall. In the UK, the needs of adolescent patients have become more widely recognised in the past decade, but they continue to have a low priority on the national agenda. Recent guidelines on cancer referral made specific reference to children's cancers, but none to cancer in adolescents. The need for dedicated services for these patients has been accepted, but the resources to meet this need have rarely been identified. We explore current and future patterns of care and service provision for adolescents with cancer in the UK. PMID- 11905620 TI - Panax ginseng--a non-organ-specific cancer preventive? AB - For the past 50 years, the main weapons in the war against cancer have been early detection and surgical removal, radiotherapy, chemotherapy, and attempts to develop gene therapy. However, the results so far are less than ideal. One strategy now is to switch from therapeutic approaches to prevention of cancer by improving lifestyle and by identifying effective natural products as chemopreventive agents. One promising candidate with cancer-preventive effects that are not specific to any organ is Panax ginseng C A Meyer, a herb with a long medicinal history. Its protective influence against cancer has been shown by extensive preclinical and epidemiological studies, but these effects need to be carefully investigated by scientific clinical trials focusing on the major cancer killers stomach, lung, liver, and colorectal cancer. PMID- 11905621 TI - DNA microarray technology to revolutionise cancer treatment. PMID- 11905622 TI - Social inequality and cancer deaths. PMID- 11905623 TI - David Zaridze--deputy director of the Russian Cancer Research Centre (interview by Sue Silver). PMID- 11905624 TI - Cancer cases on the rise in India. PMID- 11905626 TI - Australian company buys rights to Tonga gene pool. PMID- 11905625 TI - Blazing the trail. PMID- 11905627 TI - Informed consent for cancer registration. PMID- 11905628 TI - Informed consent for cancer registration. PMID- 11905629 TI - The dangers of subgroup analysis. PMID- 11905630 TI - Informed consent for cancer registry data collection. PMID- 11905631 TI - Supercomputer-designed drug protects against chemotherapy toxicity. PMID- 11905632 TI - Challenging cancer in developing countries. PMID- 11905633 TI - Dilemmas in cancer care. PMID- 11905634 TI - Alternative cancer cures. PMID- 11905635 TI - Funding for basic cancer research. PMID- 11905636 TI - STI571: an inhibitor of the BCR-ABL tyrosine kinase for the treatment of chronic myelogenous leukaemia. AB - The deregulated tyrosine kinase activity of the BCR-ABL fusion protein has been established as the causative molecular event in chronic myelogenous leukaemia. Thus, the BCR-ABL tyrosine kinase is an ideal target for pharmacological inhibition. STI571 (formerly CGP57148B), is an ABL-specific inhibitor of tyrosine kinase that, in preclinical studies, selectively killed BCR-ABL-containing cells in vitro and in vivo. Clinical studies have shown the potential of this specifically targeted therapy, and STI571 is emerging as an important new therapeutic agent for chronic myelogenous leukaemia. PMID- 11905637 TI - Lymphatic entrapment of tumour cells after sentinel lymph-node biopsy for melanoma. PMID- 11905638 TI - Photodynamic therapy: a clinical reality in the treatment of cancer. AB - Photodynamic therapy (PDT) is a minimally invasive treatment with great promise in malignant disease. It can be applied before, or after, chemotherapy, ionising radiation, or surgery, without compromising these treatments or being compromised itself. Unlike radiotherapy and surgery, it can be repeated many times at the same site. Response rates and the durability of response with PDT are as good as, or better than, those with standard locoregional treatments. Furthermore, there is less morbidity and better functional and cosmetic outcome. PDT is valuable for premalignant conditions such as mucosal dysplasia and carcinoma-in-situ. The excellent cosmetic outcome makes it valuable for skin lesions and for lesions of the head, neck, and oral cavity, where another advantage is that it has negligible effects on underlying functional structures. With endoscopic delivery of light to hollow structures, PDT has been successful in the treatment of early gastrointestinal cancers, such as oesophageal cancer, and lung cancer. The superficial effects of PDT can be exploited in the treatment of large areas such as the pleura and peritoneum, where curative radiation doses cannot be tolerated by underlying normal tissue. PDT is an ideal adjuvant therapy when surgical resection of solid tumours might leave behind residual microscopic disease. Interstitial light delivery, where light is fed directly into solid tumours, allows PDT to be used for large, buried tumours that would otherwise require extensive surgical resection. PMID- 11905639 TI - Familial colorectal cancer: pathology and molecular characteristics. AB - Appropriate management of familial colorectal cancer revolves around the diagnosis of the underlying genetic syndrome. This necessitates an interdisciplinary approach allowing integration of clinical, morphological, and molecular evidence that may involve several members of the same family. Genetic disorders express themselves over time, whereas clinical investigation of family members is likely to be episodic. Generic features of hereditary colorectal cancer syndromes include a positive family history, early age at onset, multiple neoplasms, and extracolonic lesions of either a developmental or neoplastic nature. Deriving a complete description of a genetic disorder is hampered by the need to trace and obtain tissue samples from many institutions. This review examines the usefulness of tissue-based investigations, both morphological and molecular, in raising the suspicion of familial colorectal cancer, providing a definitive tissue diagnosis and contributing to the larger body of diagnostic evidence. The account focuses on the two most well-studied syndromes--familial adenomatous polyposis (FAP) and hereditary non-polyposis colorectal cancer (HNPCC)--but consideration is also given to less well-understood syndromes. Some of these, notably hyperplastic polyposis and mixed polyposis, may closely mimic FAP or HNPCC. PMID- 11905640 TI - Haemopoietic stem-cell transplantation: recent progress and future promise. AB - Transplantation of haemopoietic stem cells is an increasingly important approach in the management of malignant haematological disease. Recent developments in our understanding of stem-cell biology have profoundly influenced the practice of both autologous and allogeneic stem cell transplantation. The demonstration that cytokines, such as granulocyte colony-stimulating factor, mobilise large numbers of haemopoietic progenitors has resulted in the peripheral blood rather than the bone marrow becoming the preferred source of haemopoietic stem cells in autologous, and increasingly in allogeneic, stem-cell transplantation. This has substantially reduced the morbidity of autografting, so that disease relapse now represents the most important cause of treatment failure. Current efforts are aimed at reducing this risk, either by employing novel conditioning regimens or by tumour purging. With regard to allogeneic transplantation, there is a growing realisation of the importance of a graft-versus-leukaemia effect, and this has encouraged the use of strategies which optimise an immunologically-mediated antitumour effect. This, coupled with increased understanding of the biology of stem-cell engraftment, has resulted in the development of less toxic conditioning regimens, designed to allow the benefits of allografting to be extended to patients in whom this procedure is contraindicated. PMID- 11905642 TI - Alan Coates--CEO of the Australian Cancer Society. Interviewed by Sue Silver. PMID- 11905641 TI - The resistance phenotype in the development and treatment of cancer. AB - Phenotypic resistance, acquired early in carcinogenesis, has an established role in the pathogenesis of cancer in well-characterised experimental systems, and possibly also has a role in the origin of human cancer. It has been suggested that sunlight, an established risk factor for human skin carcinogenesis, is able to induce rare altered cells resistant to toxicity and to favour their clonal expansion via toxic effects exerted on normal keratinocytes. Other major risk factors for human neoplasia, including smoking and ageing, may also act partly through imposition of a constrained growth environment in the target organ to favour the emergence of altered resistant cells. Strategies aimed at counteracting this constrained environment could be effective in attenuating the force that sustains clonal expansion of altered cells. PMID- 11905645 TI - Publishing clinical trial data--a long and winding road. PMID- 11905646 TI - Indian gene therapy for oral cancer. PMID- 11905643 TI - Oncology and the media. PMID- 11905647 TI - Breast cancer studies challenge tamoxifen therapy. PMID- 11905648 TI - Epidemiology of oral cancers. PMID- 11905650 TI - Ethics of genetic testing for cancer predisposition. PMID- 11905649 TI - Trends in prostate cancer mortality in England, Wales, and the USA. PMID- 11905651 TI - Cachexia and asthenia in cancer patients. AB - Over the past 10 years, there have been major advances in the understanding of cancer cachexia and asthenia. These common complications of cancer are now thought to be the consequences of complex interactions between host, tumour, and psychosocial factors. Cachexia and asthenia commonly coexist, but they can occur independently of each other. Recently identified tumour-derived factors cause lipolysis and protein catabolism. Cytokines produced by the host in response to tumour presence cause metabolic abnormalities, which result in decreased protein and lipid synthesis, increased lipolysis, and anorexia. Many other factors contribute to asthenia, such as anaemia, autonomic failure, and muscular abnormalities. Future research should clarify optimum management. The way forward seems to lie in a multidimensional approach with combined therapy to manage both cancer cachexia and asthenia. PMID- 11905652 TI - A metastatic dormant tumour in the brain. PMID- 11905653 TI - Conditionally replicative adenoviral vectors for cancer gene therapy. AB - During the past century, many attempts have been made to exploit the ability of some viruses to infect and destroy cancer cells. Crippled, non-replicative viruses have been used as vectors to transfer genes into tumours. Both strategies have serious limitations. The time is now ripe, however, for full convergence of these two research tracks. On the one hand, the intratumoral propagation of replicative viruses would overcome the low levels of gene transfer achieved by current viral vectors. On the other hand, the versatility provided by vectors encoding foreign genes, which are limited in their uses only by our ingenuity, would overcome the physiological barriers to robust propagation of the viral progeny in the tumour. This empowering synthesis will provide truly new opportunities that might realise the promises of gene transfer for the therapy of cancer. PMID- 11905654 TI - Immunotherapy for colorectal cancer: a challenge to clinical trial design. AB - Colorectal cancer is a leading cause of morbidity and mortality with about 18 000 deaths in the UK and 56 000 in the USA each year. Despite improvements in surgical technique, radiotherapy, and adjuvant chemotherapy, 50% of patients apparently 'cured' by surgery subsequently relapse and die of the disease. There is an obvious need to develop novel rational therapies based on understanding the molecular signatures that separate tumour from normal colorectal epithelium. PMID- 11905655 TI - Effect of the breast-cancer resistance protein on atypical multidrug resistance. AB - Simultaneous resistance of malignant cells to several antineoplastic agents that are structurally and functionally unrelated is known as multidrug resistance. It is one of the main causes of chemotherapy failure. Besides the classic multidrug resistant phenotype, mediated by increased activity of the ATP-binding cassette (ABC) transporter P-glycoprotein, there are other multidrug-resistant tumours, with resistance caused by different mechanisms. This is called atypical multidrug resistance. Pronounced overexpression of a novel ABC transporter has been observed in various human cancer cell lines with atypical multidrug resistance (which were established by in vitro exposure to mitoxantrone, topotecan, doxorubicin, or bisantrene). This novel transporter was originally named breast cancer resistance protein (BCRP). BCRP is a 655-aminoacid protein of about 72 kDa. It can be thought of as an ABC 'half-transporter', and it forms dimers to produce an active transport complex. Transfection experiments with BCRP cDNA showed that the phenotype of atypical multidrug resistance could be transferred to formerly drug-sensitive cancer cells. Although the role of BCRP in drug resistance of clinical cancers is still unclear, preliminary data obtained by mRNA and protein expression analyses support the assumption that it has a role in clinical multidrug resistance. PMID- 11905656 TI - The role of complementary and alternative medicine in cancer. AB - Many cancer patients use complementary and alternative medicines. Oncologists therefore need to learn more about this subject. Dietary treatments, herbal medicinal products, and various food supplements are currently being promoted as 'cancer cures', but none of these treatments are backed up by convincing clinical evidence. For some interventions, however, the evidence is sufficiently encouraging to warrant further research. Several forms of complementary and alternative medicine play a part in supportive care for cancer patients. The evidence for this approach is still preliminary but, generally speaking, promising. The challenge for the future is to obtain reliable evidence and to identify those approaches that do more good than harm. PMID- 11905658 TI - Larry Norton--president-elect of ASCO. American Society of Clinical Oncology. Interviewed by Ezzie Hutchinson. PMID- 11905657 TI - Signal transduction pathways: targets for chemoprevention of skin cancer. AB - Chemoprevention can be defined as the use of substances to interfere with the process of cancer development. Although substantial progress has been made in elucidating the basis of carcinogenesis, further advances are needed to identify molecular and cellular targets for effective use of chemopreventive agents. Hundreds of compounds have been identified as potential chemopreventive agents. However, the safety and efficacy of each substance must be thoroughly investigated. Carcinogenesis is a multistage process in which numerous genes are affected. Many of these genes regulate important cellular functions, so they are prime targets for chemopreventive agents. A major focus of our work has been the elucidation of mechanism(s) explaining the anticancer actions attributed to several chemopreventive compounds, especially 'natural compounds' that are considered safe because they are present in commonly consumed foods and beverages. Of particular interest are selected drugs (eg aspirin) and certain dietary factors (eg green and black tea, resveratrol) and their influence on cell signalling events coinciding with skin cancer promotion. This overview describes recent work from our laboratory and others focusing on molecular mechanisms of selected chemopreventive compounds in growth-related signal transduction pathways and skin cancer. PMID- 11905659 TI - Knowing the enemy. PMID- 11905660 TI - Differentiation and apoptosis induction therapy in acute promyelocytic leukaemia. AB - Induction of differentiation and/or apoptosis is a new and promising approach to cancer therapy, well illustrated by the treatment of acute promyelocytic leukaemia with all-trans-retinoic acid and arsenic compounds. Treatment with all transretinoic acid results in complete remission in 92 - 95% of patients with this disease. Using the recently advocated combination of all-transretinoic acid and chemotherapy, adverse effects such as retinoic acid syndrome have decreased, and long-term survival has improved. Chemotherapy in combination with all-trans retinoic acid seems to be the best postremission treatment protocol, with a 5 year relapse-free survival rate of 50 - 60%. Arsenic compounds have recently proved effective in newly diagnosed and relapsed acute promyelocytic leukaemia, with complete remission rates of 80 - 90% according to most reports. As2O3, the most studied arsenic compound, can be given by intravenous infusion at a dose of 0.08 - 0.16 mg/kg daily. A course of 28 - 44 days is required to induce remission. Although the drug is safe in patients who have relapsed, severe liver damage has been observed in some newly diagnosed patients. Combined use of chemotherapy and arsenic as postremission treatment results in longer survival than arsenic alone. Although their mechanisms of action are distinct, both all trans-retinoic acid and arsenic can modulate PML-RARalpha, an oncoprotein that has a central role in leukaemogenesis, and both can relieve ranscriptional repression by modifying chromatin structure. PMID- 11905661 TI - Therapy monitoring using FDG-PET in metastatic cervical cancer. PMID- 11905662 TI - Olive-oil consumption and health: the possible role of antioxidants. AB - In the Mediterranean basin, olive oil, along with fruits, vegetables, and fish, is an important constituent of the diet, and is considered a major factor in preserving a healthy and relatively disease-free population. Epidemiological data show that the Mediterranean diet has significant protective effects against cancer and coronary heart disease. We present evidence that it is the unique profile of the phenolic fraction, along with high intakes of squalene and the monounsaturated fatty acid, oleic acid, which confer its health-promoting properties. The major phenolic compounds identified and quantified in olive oil belong to three different classes: simple phenols (hydroxytyrosol, tyrosol); secoiridoids (oleuropein, the aglycone of ligstroside, and their respective decarboxylated dialdehyde derivatives); and the lignans [(+)-1-acetoxypinoresinol and pinoresinol]. All three classes have potent antioxidant properties. High consumption of extra-virgin olive oils, which are particularly rich in these phenolic antioxidants (as well as squalene and oleic acid), should afford considerable protection against cancer (colon, breast, skin), coronary heart disease, and ageing by inhibiting oxidative stress. PMID- 11905663 TI - Professor Sir Michael Rawlins--chairman of NICE. National Institute of Clinical Excellence. Interviewed by Sue Silver. PMID- 11905664 TI - Genetic testing for cancer predisposition--an ongoing debate. PMID- 11905665 TI - Clinical trials and RIRO. PMID- 11905666 TI - The balance between care and cure. PMID- 11905667 TI - US cancer charity launches campaign to change American eating habits. PMID- 11905668 TI - The sky's the limit for new drug design centre. PMID- 11905669 TI - Aggressive combination therapy to cure patients with metastatic cancer. PMID- 11905670 TI - Mode of action of farnesyltransferase inhibitors. PMID- 11905671 TI - Advances in IMRT: a clinical perspective. PMID- 11905672 TI - Management of soft-tissue sarcomas: an overview and update. AB - Soft-tissue sarcomas (STS) are relatively uncommon, especially when considered as individual histological subtypes (of which there are more than 50). Their incidence increases with age, although they are disproportionately common among children. When diagnosed and managed in a non-specialist environment, outcome is generally significantly poorer than if patients are managed by a multidisciplinary team in a tertiary centre of excellence. Prompt referral of patients with clinically suspicious masses is strongly advocated, before any type of intervention is attempted. This brief, opinion-based overview emphasises the team approach and provides a synopsis of the strategies used at our institution for pre-operative assessment and biopsy, surgical management, and the delivery of radiation therapy when appropriate (focusing on limb preservation and optimisation of function). Predictable variations in the natural history of these tumours, based on accurate histological subclassification, merit wider recognition. The role of systemic chemotherapy for soft-tissue sarcoma is still evolving, but at present the main aims are improved local control, disease-free survival, and quality of life. There are overall survival benefits for specific histological types, but this is a relatively small subgroup. Novel therapies, based on disease mechanisms at the molecular level, show promise for future advances. PMID- 11905673 TI - Repopulation of tumour cells between cycles of chemotherapy: a neglected factor. AB - Repopulation of clonogenic tumour cells during fractionated radiation treatment is recognised as an important factor affecting local control. Given the longer intervals between cycles and longer total duration of treatment, the impact of repopulation is likely to be greater following chemotherapy. Limited data from experimental models suggest that, after chemotherapy, there is a 'lag period', followed by variable but rapid rates of repopulation of tumour cells, possibly accelerating between cycles. Modelling of these properties indicates that after the initial response, accelerated repopulation between cycles can lead to tumour regrowth without any change in the drug sensitivity of the tumour cells. The importance of repopulation may be comparable with that of intrinsic or acquired cellular resistance in determining the effective resistance of tumours to chemotherapy. Biological agents with rapid onset and short duration of action, which can selectively inhibit tumour-cell repopulation, administered between cycles of chemotherapy, might improve the therapeutic index. PMID- 11905674 TI - SIOPEL trials using preoperative chemotherapy in hepatoblastoma. AB - Hepatoblastoma is a rare, malignant liver tumour of childhood. Until the mid 1980s only around 30% of patients were cured, but with modern chemotherapy, and of course surgery, the cure rate is now at least 70%. This dramatic improvement in survival has been achieved by the national and international cooperation of paediatric oncology centres. The International Society of Paediatric Oncology Liver Tumour Group, in contrast to most other groups, has used preoperative chemotherapy in all patients, followed by delayed surgery. The group has also developed a novel staging system, called PRETEXT (PRE Treatment EXTent of disease), based on the anatomy of the liver and radiological findings at diagnosis, to try to predict resectability and outcome. PMID- 11905676 TI - Hereditary cancer. PMID- 11905675 TI - Ray of light for treatment of head and neck cancer. PMID- 11905677 TI - PSA assays. PMID- 11905678 TI - Trends in prostate cancer mortality in England, Wales, and the USA. PMID- 11905679 TI - PSA doubling times. PMID- 11905680 TI - Importance of field cancerisation in clinical oncology. PMID- 11905681 TI - Quantitating quality of life. PMID- 11905682 TI - Prostate cancer: to screen or not to screen? AB - The aim of screening is to identify cancers that are potentially curable; before a programme can be introduced, it must satisfy the requirement that it does more good than harm, particularly in terms of survival and quality of life. Prostate cancer is a common disease in older men and presents a significant burden to health services. Prostatic tumours range from small slow-growing lesions to aggressive tumours that metastasise rapidly, but because the natural history of prostate cancer is poorly understood, there is controversy about which screen detected lesions will become clinically significant. Current methods of screening involve measurement of serum prostate specific antigen, followed by transrectal ultrasound scanning and biopsy, but these lack adequate specificity and sensitivity. There are three major treatment options for localised disease: radical prostatectomy, radical radiotherapy, and monitoring with treatment if required. There is no randomised controlled trial evidence to suggest a survival advantage of any of these treatments, and each has risks. There is intense speculation about future developments in diagnostic testing, molecular markers of progression, and early chemoprevention, but the central question that remains is whether radical treatments can improve survival and quality of life. PMID- 11905683 TI - Cutaneous B cell lymphoma arising from a chronically inflamed sebaceous cyst. PMID- 11905685 TI - A shot in the dark. PMID- 11905684 TI - The role of hypoxia-activated prodrugs in cancer therapy. AB - Tumour hypoxia, a deficiency of oxygen due to an inefficient vasculature, is a limiting factor in both the radiotherapy and chemotherapy of solid tumours. Paradoxically, it is also an attractive therapeutic target, because severe hypoxia occurs only in solid tumour tissue. Hypoxic cells can be exploited for therapy by non-toxic, hypoxia-activated prodrugs. Conceptually, 'trigger' units in these drugs are selectively activated in hypoxic cells to release or activate a toxic 'effector', capable of killing surrounding oxygenated tumour cells. Useful triggers include nitroaromatics, quinones, N-oxides, and transition metals. The N-oxide tirapazamine is in phase III clinical trials. PMID- 11905686 TI - Advances in three-dimensional conformal radiation therapy physics with intensity modulation. AB - Intensity-modulated radiation therapy, a specific form of conformal radiation therapy, is currently attracting a lot of attention, and there are high expectations for this class of treatment techniques. Several new technologies are in development, but physicists are still working to improve the physical basis of radiation therapy. PMID- 11905687 TI - Ethnicity and oral cancer. AB - Oral squamous-cell carcinoma, the main type of oral cancer, is among the ten most common cancers in the world. The aims of this paper were first, to consider whether there was evidence of marked ethnic variations in the incidence, management, and survival of oral cancer, and then, to review possible explanations for these variations. Evidence from the literature suggests that there is marked, inter-country variation in both the incidence and mortality from oral cancer. There is also growing evidence of intracountry ethnic differences, mostly reported in the UK and USA. These variations among ethnic groups have been attributed mainly to specific risk factors, such as alcohol and tobacco (smoking and smokeless), but dietary factors and the existence of genetic predispositions may also play a part. Variations in access to care services are also an apparent factor. The extent of ethnic differences in oral cancer is masked by the scarcity of information available. Where such data are accessible, there are clear disparities in both incidence and mortality of oral cancer between ethnic groups. PMID- 11905688 TI - Tamoxifen: a personal retrospective. AB - Tamoxifen, originally described as an anti-oestrogen and antifertility agent in the rat, is now a pioneering medicine for the treatment and prevention of breast cancer. Its success is the result of an effective collaboration between laboratory research and clinical trial processes. However, this drug is more than just an anti-oestrogen to treat breast cancer. Laboratory and clinical research defined the concept of selective oestrogen receptor modulation in the 1980s. Non steroidal anti-oestrogens show oestrogen-like activity in bones and lower cholesterol, but block oestrogen action in the breast and uterus. This realisation led to the development of chemical cousins, known as selective oestrogen receptor modulators. One of these compounds, raloxifene, is used for the prevention of osteoporosis, but is currently being tested as a preventive for breast cancer. PMID- 11905689 TI - Proteomics may aid bladder cancer diagnosis. PMID- 11905690 TI - Axel Ullrich--a pioneer in gene technology. Interviewed by Ezzie Hutchinson. PMID- 11905691 TI - 'Alternative cancer cures': looking for common ground. PMID- 11905692 TI - Plumbing the human genome. PMID- 11905693 TI - Better prospects for myeloma patients. PMID- 11905694 TI - New virtual computer on trial. PMID- 11905695 TI - Potent anti-cancer drug from witches' umbrella. PMID- 11905696 TI - Indian centre develops teletherapy machine for cervical cancer. PMID- 11905697 TI - Melanoma vaccine launch may be too early. PMID- 11905698 TI - Embryonic stem cell research--still at the starting gate. PMID- 11905699 TI - Spain hopes to enter big league in oncology research. PMID- 11905700 TI - New bisphosphonate shows promise in bone metastases. PMID- 11905701 TI - Generic drugs to increase access to chemotherapy in Chile. PMID- 11905702 TI - Johns Hopkins and RCC face drug trial allegations. PMID- 11905703 TI - Pill-sized camera seeks out intestinal cancers. PMID- 11905704 TI - The clinical investigator award--promoting patient-related research. PMID- 11905705 TI - Public and private money needed to stop 'physician brain drain'. PMID- 11905706 TI - Euthanasia: to kill or not to kill? PMID- 11905707 TI - Global cancer statistics in the year 2000. AB - Estimation of the burden of cancer in terms of incidence, mortality, and prevalence is a first step to appreciating appropriate control measures in a global context. The latest results of such an exercise, based on the most recent available international data, show that there were 10 million new cases, 6 million deaths, and 22 million people living with cancer in 2000. The most common cancers in terms of new cases were lung (1.2 million), breast (1.05 million), colorectal (945,000), stomach (876,000), and liver (564,000). The profile varies greatly in different populations, and the evidence suggests that this variation is mainly a consequence of different lifestyle and environmental factors, which should be amenable to preventive interventions. World population growth and ageing imply a progressive increase in the cancer burden--15 million new cases and 10 million new deaths are expected in 2020, even if current rates remain unchanged. PMID- 11905708 TI - Hepatocellular carcinoma and lymphoma--two hepatitis B virus-related malignant diseases. PMID- 11905709 TI - Cyclo-oxygenase 2: a pharmacological target for the prevention of cancer. AB - Understanding the mechanisms underlying carcinogenesis provides insights that are necessary for the development of therapeutic strategies to prevent cancer. Chemoprevention--the use of drugs or natural substances to inhibit carcinogenesis - is an important and rapidly evolving aspect of cancer research. We discuss evidence that cyclooxygenase 2 (COX 2), an inducible form of the enzyme, is a potential pharmacological target to prevent cancer. Key data implicating a causal relation between increased activity of COX 2 and carcinogenesis and possible mechanisms of action of COX 2 in this context are covered. Importantly, selective COX 2 inhibitors appear to be safe enough in human beings to allow large-scale clinical testing in healthy people. Several chemoprevention trials using selective COX 2 inhibitors are underway. PMID- 11905710 TI - Current and future developments in the use of temozolomide for the treatment of brain tumours. AB - Brain tumours comprise only 2% of all adult cancers, but they are among the most debilitating malignant diseases. Temozolomide, an alkylating agent that can be administered orally, has been approved for the treatment of recurrent malignant glioma on a daily schedule for 5-day cycles. Continuous administration schedules with a higher dose intensity are being explored, but an improvement in efficiency remains to be shown. The benefit from temozolomide given as a single agent in recurrent disease will be several weeks at best. This drug is therefore now undergoing clinical testing as neoadjuvant chemotherapy or with concomitant radiotherapy in patients with newly diagnosed glioma. Several phase I trials are investigating the combination of temozolomide with other agents active against brain tumours. This review briefly summarises the pharmacological background and clinical development of temozolomide and focuses on current and future clinical exploration of this drug for the treatment of brain tumours. PMID- 11905711 TI - Preoperative/neoadjuvant medical therapy for early breast cancer. AB - Preoperative (neoadjuvant) medical therapy has emerged over the past decade as a new approach for the treatment of early breast cancer. Results show it has high activity, but survival is no better than with conventional adjuvant treatment. The need for mastectomy is reduced but not abolished; in some studies this effect is associated with a small increase in risk of local recurrence, but without any detriment to survival. Predictive factors for improved outcome include clinical response, and especially pathological complete remissions. However, persisting pathological axillary node involvement is associated with poor outcome. Biological changes in apoptosis or proliferation pathways may prove to be more sensitive surrogate markers than clinical or pathological responses for assessing treatment outcome. The main long-term aim of preoperative medical treatment must be to establish such surrogate predictive markers. This would lead to individualised treatment for each patient, and would allow much more rapid assessment of new drugs than is currently possible with adjuvant therapy trials. PMID- 11905713 TI - Sylvia Denton: breast care nurse (interview by Ezzie Hutchinson). PMID- 11905712 TI - International myeloma grand round. AB - On the 14th June 2001, The Royal Marsden Hospital, London, hosted an unusual event, an international grand round, aimed at exploring a new concept in the treatment of multiple myeloma--operational cure. The speakers were Jean-Luc Harousseau (France), Gregory Mundy (USA), Brian Durie (USA) and Ray Powles (UK). Case histories were presented by Bhawna Sirohi (UK). The 80 invited guests included oncologists, radiotherapists, histopathologists, representatives of the International Myeloma Foundation, members of the press, and several myeloma patients, including the two whose case histories are presented below. PMID- 11905714 TI - A golden rule. PMID- 11905716 TI - Safer cigarettes anger anti-smoking campaigners. PMID- 11905715 TI - Unresolved crisis in clinical research. PMID- 11905717 TI - Palliative care in the USA critised. PMID- 11905718 TI - Wandering brachytherapy seeds need to be monitored. PMID- 11905719 TI - Raloxifene encourages growth of ovarian cancer cells in vitro news. PMID- 11905720 TI - The use of dubious cell lines in research: is trust enough? PMID- 11905721 TI - Cell banks detect false cell lines: journals must act too. PMID- 11905722 TI - Optimum anthracycline-based chemotherapy for early breast cancer. AB - Adjuvant chemotherapy improves the overall survival of women treated after surgery for early breast cancer. Several trials have suggested that anthracycline containing regimens are more effective than those that do not contain anthracyclines. A modest overall benefit has also been confirmed by the Early Breast Cancer Trialists' Collaborative Group overview. Newer agents, such as the taxanes, are now being tested in the adjuvant setting in randomised trials. The control group of such studies should receive the optimum standard treatment. There are several anthracycline-based regimens in common use, varying in terms of the type of anthracycline used, the dose, and drug scheduling. We review the available evidence and consider whether the optimum anthracycline-containing chemotherapy schedule has now been identified. PMID- 11905723 TI - Meta-analyses of randomised clinical trials in oncology. AB - A meta-analysis is a quantitative synthesis of randomised clinical trials, used to evaluate moderate treatment effects in oncology. It is complementary to large scale trials. We describes the principles, methods, and limits of meta-analyses. The gold standard for a meta-analysis is to obtain individual patient data directly from each principal investigator, but this is time-consuming and costly. The main steps of a meta-analysis using individual patient data are described. Multidisciplinary collaboration is needed for clinical insight and critical review of the data and results. Meta-analysis should include an evaluation of the trial quality, a quantification of the overall treatment effect, a study of the variations seen in this effect between trials, and pre-planned exploratory analyses to identify groups of patients who may benefit more from the treatment. Statistical methods are explained using real working examples. Since literature based meta-analysis can lead to seriously biased assessments, meta-analyses of individual patient data should be undertaken systematically when long-term follow up is needed, when a detailed analysis is important, or when the literature-based meta-analyses are not in agreement. The main factors which influence the quality of a meta-analysis are discussed. PMID- 11905724 TI - Repair of DNA interstrand crosslinks: molecular mechanisms and clinical relevance. AB - Drugs that produce DNA interstrand crosslinks (ICLs), between the two complementary strands of the double helix, have an important role in chemotherapy regimens for cancer. Novel crosslinking agents, and targeting strategies involving DNA crosslinking agents, continue to be developed. The ability of cells to repair DNA ICLs is a critical determinant of sensitivity, and recent dinical studies indicate that DNA repair capacity is strongly implicated in both inherent tumour sensitivity and acquired drug resistance. A detailed understanding of the cellular mechanisms that act to eliminate these critical DNA lesions is clearly important. DNA ICLs present a complex challenge to DNA repair mechanisms because of the involvement of both DNA strands. It is now clear that cells from bacteria and yeast to mammals eliminate interstrand ICLs through the coordinated action of several DNA repair pathways. Recently, a model of ICL repair has been proposed, in which mammalian cells use novel excision repair reactions (requiring the XPF and ERCC1 proteins) to uncouple the crosslink. This is followed by a homologous recombination step to provide the genetic information needed to complete repair. This new knowledge may permit the development of screens for tumour response to crosslinking agents, and should also aid the design of more effective crosslinking agents that evade DNA repair. In addition, the proteins mediating the repair reactions represent potential targets for therapeutic intervention. PMID- 11905725 TI - Immunotherapy of cancer with alloreactive lymphocytes. AB - Immunotherapy of cancer with alloreactive lymphocytes is the mainstay of treatment, especially in haematological malignant disease. With donor lymphocyte infusion for immunotherapy, it is essential to induce host-versus-graft tolerance to ensure that the donor lymphocytes are accepted. Engraftment of haemopoietic cells of donor origin can be accomplished with reduced-intensity conditioning. Reducing transplant-related mortality by simplifing the stem-cell transplant procedure with a reduced-intensity regimen, particularly non-myeloablative conditioning, may have great potential for the treatment of malignant and non malignant disorders. PMID- 11905726 TI - Depression in cancer patients. AB - Clinical depression is a relatively common, and yet frequently overlooked, source of suffering among patients with cancer. All patients who face a life-threatening diagnosis such as cancer experience a normal albeit painful emotional reaction, but a substantial minority will become clinically depressed. This article reviews some basic information that oncology practitioners may find helpful in identifying patients at risk of experiencing a major depression. A brief overview of the epidemiology, diagnostic criteria, screening approaches, and special issues, such as depression in the elderly, high-risk populations, and suicide is also provided. PMID- 11905728 TI - Emil J Freireich: a pioneer of modern oncology (interview by Sue Silver). PMID- 11905727 TI - Women and lung cancer: does oestrogen play a role? AB - Smoking-related disease remains a major public-health problem. Large numbers of women continue to smoke, and new smokers are almost as likely to be female as male. Lung cancer is still a largely incurable disease; annual lung-cancer mortality in women exceeds that of breast cancer, and lung cancer now accounts for 12% of all new female cancer cases. The results of several studies suggest that women are more susceptible than men to lung cancer and to conditions that predispose to this cancer, such as chronic obstructive pulmonary disease. There is still much controversy about whether there is an increased lung-cancer risk in women across all populations. Many epidemiological studies have been negative or equivocal when comparing male and female lung-cancer risk. This article is not intended to be a comprehensive review of all epidemiological studies, or of all possible lung-cancer risk factors. Lung-cancer incidence and risk in women are discussed, and evidence for possible mechanisms of increased female risk are presented, including the role of oestrogen in the development of lung cancer. PMID- 11905729 TI - Shaky foundations. PMID- 11905730 TI - Contamination of cell lines--a conspiracy of silence. PMID- 11905731 TI - The keyhole approach to kidney removal. PMID- 11905732 TI - St John's wort helps to fight bladder cancer. PMID- 11905733 TI - Capecitabine monotherapy in metastatic colorectal cancer. PMID- 11905734 TI - Programming of radiotherapy in the treatment of non-small-cell lung cancer--a way to advance care. AB - Radical radiotherapy, the mainstay of treatment for early inoperable non-small cell lung cancer, is most commonly given in daily fractions, Monday to Friday, to a total dose of 60-70 Gy over 6-8 weeks. Since the 1980s, novel fractionation schedules have been explored with the aim of improving local tumour control and survival without increasing late morbidity. There have been two main approaches. In hyperfractionated radiotherapy the dose per fraction is reduced and the total dose increased to give improved tumour control without increased late morbidity. Hyperfractionation schedules, with more than one fraction per day have been successfully evaluated, but so far significant benefit has not been achieved when compared with conventional radiotherapy plus chemotherapy. In accelerated radiotherapy the overall duration of radiotherapy is reduced to overcome repopulation of tumour cells during the course of treatment. In all the different regimens of accelerated radiotherapy a common feature is giving two or more fractions on some or all treatment days and, in some cases, a lower dose per fraction is also incorporated. CHART (continuous hyperfractionated accelerated radiotherapy) is the most novel and accelerated schedule tested, and a randomised controlled trial showed a significant survival advantage from CHART compared with conventional radiotherapy. Changes in the fractionation of radiotherapy must be combined with other approaches such as neoadjuvant and concomitant chemotherapy, hypoxic-cell modifiers, and conformal radiotherapy, so that care of patients with non-small-cell lung cancer can be further advanced. PMID- 11905735 TI - Minimal residual disease in leukaemia patients. AB - Because of developments in diagnosis of haemopoietic malignant diseases during the past two decades, routine and reliable identification of very low numbers of malignant cells, known as minimal residual disease (MRD), is now possible. Several large-scale studies have shown that monitoring of MRD in haemopoietic malignant disease predicts clinical outcome. In acute lymphoblastic leukaemia, MRD detection is useful for evaluating early response to treatment and consequently for improving stratification, including treatment reduction. In acute promyelocytic leukaemia and chronic myeloid leukaemia, MRD information at specific time points enables effective early treatment intervention. MRD monitoring is also possible in other leukaemia subtypes, but in these disorders the clinical value of MRD detection is not yet known. PMID- 11905736 TI - Hepatic-arterial chemotherapy. AB - The liver is a common site of metastases from cancers from most sites, but particularly from the gastrointestinal tract, since the portal vein drains into the liver. About half of all patients with colorectal cancer develop liver metastases. The response of liver metastases to systemic combination chemotherapy has improved, but the 2-year survival is only 25-30%. Hepatic-arterial infusion of chemotherapy produces higher response rates, with a 2-year survival of 50-60%. In patients who can undergo liver resection followed by hepatic-arterial infusion, the 2-year survival is 85%. This review summarises the anatomical basis, pharmacokinetic background, and cost-effectiveness of this procedure. We discuss the phase II and phase III studies of hepatic-arterial infusion therapy, with a focus on liver metastases from colorectal cancer. PMID- 11905738 TI - The role of extracellular matrix in small-cell lung cancer. AB - Lung cancer is the most common fatal malignant disease in the western world, accounting for 42,000 deaths each year in the UK alone. Small-cell lung cancer (SCLC), accounts for 25% of all lung cancers. It is a particularly aggressive form of the disease, characterised by widespread metastases and the development of resistance to chemotherapy. Even with combination chemotherapy and radiotherapy treatments, the 5-year survival is only about 5%. We review recent insights into the mechanisms underlying the development of metastases and resistance to chemotherapeutic agents in SCLC, focusing on the role of the extracellular matrix (ECM). We discuss the regulation of the interactions between cells and the ECM and the effects of these interactions on cellular phenotypes, together with some of the new approaches for combating drug resistance and metastases in this disease. PMID- 11905737 TI - Pattern of malignant disorders in individuals with Down's syndrome. AB - The pattern of occurrence of malignant disorders in people with Down's syndrome (DS) is unique and may serve as a model in the search for leukaemogenic genes and tumour suppressor genes on chromosome 21, since the risk of leukaemia is higher in individuals with DS than in non-DS individuals. Acute lymphoblastic leukaemia in DS shares many of the clinical characteristics of the same malignancy in other patients, and with current intensive therapy the long-term survival is similar. Myelodysplastic syndrome and acute myeloid leukaemia have unique clinical characteristics in these patients and are best described as a single disorder, termed myeloid leukaemia of DS. When these patients are treated intensively, they show better survival rates than patients without DS. This may be related to increased expression of genes on chromosome 21 contributing to increased chemosensitivity. Chronic myeloid leukaemia and chronic lymphocytic leukaemia occur less often than expected. With the exception of an increased risk of retinoblastoma, germ-cell tumours, and perhaps lymphomas, the risk of developing solid tumours is lower in both children and adults. Breast cancer is almost absent, and the risk of a second malignant disease after treatment for leukaemia also appears to be decreased. Increased susceptibility to apoptosis in DS may result in cell death rather than malignant transformation after major cell injuries. This hypothesis would explain the decreased risk of both solid tumours and secondary cancers. PMID- 11905739 TI - Dermoscopy of pigmented skin lesions--a valuable tool for early diagnosis of melanoma. AB - The clinical use of dermoscopy has uncovered a new and fascinating morphological dimension of pigmented skin lesions. Dermoscopy is a non-invasive diagnostic technique that links clinical dermatology and dermatopathology by enabling the visualisation of morphological features not seen by the naked eye. Close examination of pigmented skin lesions in this way increases the effectiveness of clinical diagnostic tools by providing new morphological criteria for distinguishing melanoma from other melanocytic and non-melanocytic pigmented skin lesions. In the past, dermoscopy has been known by various names, including skin surface microscopy, epiluminescence microscopy, incident light microscopy, dermatoscopy, and videodermatoscopy. However, the term 'dermoscopy', first used by Friedman and colleagues in 1991, is the most widely used. PMID- 11905741 TI - A sense of context. PMID- 11905740 TI - Chun Wang on oncology in China. PMID- 11905742 TI - Genetic testing--are we ready? PMID- 11905743 TI - A new alternative for prostate cancer treatment in Spain. PMID- 11905744 TI - Major campaign to raise colon cancer awareness in Germany. PMID- 11905745 TI - NASA and NCI join forces to work on biomolecular sensors. PMID- 11905746 TI - Immunotherapy shows promise in Hodgkin's disease. PMID- 11905747 TI - Medical marijuana on trial. PMID- 11905748 TI - EORTC mistletoe study. PMID- 11905749 TI - Combined chemotherapy and radiation in locally advanced non-small-cell lung cancer. AB - The efficacy of radiotherapy in locally advanced non-small-cell lung cancer is limited. One attempt to improve survival uses a combination of radiation and chemotherapy. These two modalities can be applied in sequence or concurrently, but results from phase III trials of combined therapy versus radiation alone have been inconsistent. Early studies were mostly negative, but more recent trials using platinum-based regimens have shown some survival benefit for combined treatments. The positive impact of chemotherapy has also been shown in a meta analysis. In recent studies, concurrent chemotherapy and radiation appears better than sequential application. However, the benefit of the combined approach is modest and should be balanced against increased early and late toxicity. The role of new agents such as taxanes, vinorelbine, gemcitabine, and topoisomerase inhibitors in combined modality therapy of non-small-cell lung cancer warrants further clinical investigation. PMID- 11905750 TI - Brachial plexopathy affecting the development of Beau's lines unilaterally. PMID- 11905751 TI - The use of dendritic cells in cancer therapy. AB - Although the immune system evolved to protect the host from infection, what fires the popular imagination is its potential to recognise and destroy cancer. The immune system can generate potent cytotoxicity (eg transplant rejection), but can these mechanisms be harnessed for therapeutic benefit in patients with cancer? The discovery of an ever-increasing array of tumour antigens shows clearly that the targets exist. The challenge lies in generating a sufficiently potent response towards them. Central to the processes of antigen recognition, processing, and presentation to the immune system are dendritic cells. Understanding of the relation between these and the cellular immune response is crucial to elucidation of how to manipulate immune responses. The past 20 years have witnessed a dramatic expansion in this understanding and led to the first early-phase clinical trials of dendritic cells for the treatment of cancer. These studies have established the safety and feasibility of this approach and have produced encouraging evidence of therapeutic efficacy. This paper reviews the biology of dendritic cells and their use in clinical trials, as well as highlighting issues for future trial design. PMID- 11905752 TI - Primary nervous-system lymphoma. AB - Primary nervous-system lymphoma is a rare type of non-Hodgkin lymphoma, which is confined to the nervous system. This disease is managed quite differently from the usual treatment of either primary brain tumours or systemic non-Hodgkin lymphoma. Although whole-brain radiotherapy results in responses in more than 90% of cases, this treatment is associated with high relapse rates and with delayed neurotoxicity in elderly patients. First-generation chemotherapy regimens used successfully in systemic non-Hodgkin lymphoma (eg cyclo-phosphamide, adriamycin, vincristine, and prednisone) are ineffective in primary nervous-system lymphoma, partly because of the blood-brain barrier. Median survival of patients treated with radiotherapy alone or chemotherapy plus radiotherapy is similar, and ranges from 10 to 16 months. The addition of methotrexate-based chemotherapy has improved survival for these patients, extending median survival to more than 30 months. When used alone, methotrexate-based chemotherapy is associated with significantly fewer treatment-associated toxic effects. Leptomeningeal lymphoma and intraocular lymphoma are topics of particular relevance in primary nervous system lymphoma and are addressed in this review. PMID- 11905753 TI - Radiotherapy and cellular signalling. AB - Developments in cellular and molecular biology in the past decade have increased our understanding of the processes by which cells respond to ionising radiation. Cells use complex protein signalling systems that recognise radiation damage to DNA and plasma membrane lipids. When damage is found, it leads to the activation of various intracellular pathways that modulate the activity of genes controlling ceflular responses such as apoptosis, cell-cycle arrest, or repair. Numerous molecular targets may be activated or inhibited in an attempt to upregulatre or downregulate the radiation response. In this review, we discuss some of the new compounds and techniques for manipulating the cell's response to radiation. PMID- 11905755 TI - Breaking bad news--development of a hospital-based training workshop. AB - The skills required to break bad news have been written about extensively and are taught in medical schools. Recent initiatives have concentrated on improving the skills of doctors and nurses in senior positions, who act as role models for their junior colleagues. A multiprofessional learning situation can be a threatening environment, in which colleagues may worry about exposing some of the weaknesses in their knowledge and skills regarding communication with patients. We describe the initiation, running, and evaluation of successful training workshops on breaking bad news in a large British district hospital. PMID- 11905754 TI - Cancer chemoprevention by dietary constituents: a tale of failure and promise. AB - Although the results of clinical intervention trials of beta-carotene to prevent lung cancer, and of dietary augmentation with fibre or fruit and vegetables to reduce the occurrence of colonic polyps have so far been negative, a structured path for the development of diet-derived constituents as cancer chemopreventive agents is emerging. Putative agents are identified on the basis of epidemiological and preclinical mechanistic studies. Some examples of promising diet-derived chemopreventive agents are folate, curcumin, genistein, and tea catechins. Long-term supplementation of the diet with folate seems to lower the risk of colorectal cancer. Curcumin in the spice turmeric, genistein in soya, and catechins in tea have tumour-suppressing properties in rodent models of carcinogenesis, and they interfere with cellular processes involved in tumour promotion and progression. Kinases, telomerase, cyclooxygenase-2, triggers of apoptosis, and transcription factors AP1 and nuclear factor kappaB are among the cellular targets. The investigation of dietary constituents should follow a structured design, incorporating parallel preclinical studies of the food source and the isolated agent in terms of efficacy, toxicity, biological mechanisms, and pharmacokinetics. Either the food source or the isolated agent should be selected for further development on the basis of dose-efficacy and toxicity data. Pilot clinical trials on the pharmacokinetics and mechanism-based markers of efficacy of the selected intervention should precede phase I-III development in suitable populations. PMID- 11905756 TI - Edison Liu--Director of the Genome Institute of Singapore (interview by Ezzie Hutchinson). PMID- 11905758 TI - Very rare cancers--a problem neglected. PMID- 11905757 TI - Where treatment begins. PMID- 11905759 TI - New form of paclitaxel shows promise. PMID- 11905760 TI - Human genome gives academia a chance to shine. PMID- 11905761 TI - Large-scale risk assessment for 'DES daughters'. PMID- 11905763 TI - India's new smoking laws. PMID- 11905762 TI - Depleted uranium--is it really a health issue? PMID- 11905764 TI - Management of hot flashes in breast-cancer survivors. AB - Hot flashes can be a major problem for patients with a history of breast cancer. Although oestrogen can alleviate hot flashes to a large extent in most patients, there has been debate about the safety of oestrogen use in survivors of breast cancer. The decrease in hot flashes achieved with progestational agents is similar to that seen with oestrogen therapy but, again, there is some debate about the safety of progestational agents in patients with a history of breast cancer. Several alternative substances have therefore been investigated. These include a belladonna alkaloid preparation, clonidine, soy phyto-oestrogens, vitamin E, gabapentin, and several of the newer antidepressants, with venlafaxine being the best studied to date. Several studies in progress may provide better non-hormonal means of treating hot flashes in the future. PMID- 11905765 TI - Vaccines for melanoma: translating basic immunology into new therapies. AB - Advances in molecular biology and immunology in the past 10-15 years have allowed for a greater understanding of the molecules present on melanoma cells that are recognised by the immune system. The rising incidence of melanoma, combined with lack of efficacy of cytotoxic therapies, means there is a significant need for the development of effective immunotherapies. We discuss three types of vaccine for melanoma, which are currently in phase III clinical trials: allogeneic and autologous cellular vaccines, and carbohydrate vaccines. We also discuss several new areas of vaccine development, including DNA vaccines, dendritic-cell-based vaccines, peptide vaccines, and heat-shock protein vaccines. Although initial clinical trials have shown the safety and immunological efficacy of vaccines for melanoma, the true clinical benefit of these strategies will only be revealed in large randomised trials. PMID- 11905766 TI - Cancer after exposure to radiation in the course of treatment for benign and malignant disease. AB - This review assesses the patterns of radiation-associated relative risks of cancer incidence and mortality in groups exposed to ionising radiation in the course of treatment for various malignant and non-malignant conditions. In general, the relative risks among Japanese survivors of atomic-bomb explosions are greater than those among comparable subsets in studies of medically exposed individuals. Cell sterilisation largely accounts for the discrepancy in the relative risks between these two populations, although other factors may contribute, such as the generally higher underlying cancer risks in the medical series than in the Japanese atomic-bomb survivors, and dose-fractionation effects. The higher underlying cancer risk in some of the medically exposed populations, in particular for those with cancer-prone conditions, implies that the absolute excess risk is sometimes higher than in the Japanese data. For these cancer-prone individuals, the increase in absolute risk for those receiving the large doses of radiation associated with radiotherapy may be quite significant. This increase in risk has to be balanced against the generally high spontaneous cancer risk in these individuals and the benefits accruing from radiotherapy. PMID- 11905767 TI - Marine organisms as a source of new anticancer agents. AB - Various active anticancer agents are derived from plants and terrestrial microorganisms. The isolation of C-nucleosides from the Caribbean sponge, Cryptotheca crypta, four decades ago, provided the basis for the synthesis of cytarabine, the first marine-derived anticancer agent to be developed for clinical use. Cytarabine is currently used in the routine treatment of patients with leukaemia and lymphoma. Gemcitabine, one of its fluorinated derivatives, has also been approved for use in patients with pancreatic, breast, bladder, and non small-cell lung cancer. Over the past decade, several new experimental anticancer agents derived from marine sources have entered preclinical and clinical trials. This field has expanded significantly as a result of improvements in the technology of deep-sea collection, extraction, and large-scale production through aquaculture and synthesis. In this paper, examples of marine-derived experimental agents that are currently undergoing preclinical and early clinical evaluation are briefly discussed. A summary of the available information on the results of phase I and II trials of agents such as aplidine, ecteinascidin-734 (ET-734), dolastatin 10 and bryostatin 1 is also presented. PMID- 11905768 TI - Unconventional therapies for cancer and cancer-related symptoms. AB - A significant proportion of cancer patients try unconventional therapies and many use 'complementary' therapies, as adjuncts to mainstream care, for management of symptoms and to improve quality of life. A smaller proportion use 'alternative' therapies, which are typically invasive, biologically active, and commonly promoted as replacements for, rather than adjuncts to, mainstream therapy. Many alternative therapies, including high-dose vitamin C, the Di Bella regimen, and laetrile have been shown not to be effective. For others, such as metabolic therapy, evidence is extremely limited. Conversely, most complementary therapies are well studied and of proven benefit. There is evidence from randomised trials supporting the value of hypnosis for cancer pain and nausea; relaxation therapy, music therapy, and massage for anxiety; and acupuncture for nausea. Such complementary therapies are increasingly provided at mainstream cancer centres. PMID- 11905769 TI - Metastatic neoplasia involving a mitral bioprosthesis. PMID- 11905770 TI - The role of oncology nurses in gene therapy. AB - In February 2001, the Human Genome Project international consortium announced the publication of a draft sequence and initial analysis of the human genome. Although a definitive count of human genes must await further experimental and computational analysis, scientists now estimate that the human genome contains 30000-35000 genes--a much smaller number than initially estimated. The advances in treatment which will result from this research and our improved understanding of cancer at a molecular level will rapidly change the management of cancer. Gene therapy represents one new approach to treatment, but is currently still experimental. This article reviews the important role of the oncology nurse as a member of the multidisciplinary team caring for patients who receive gene therapy as part of a clinical trial. PMID- 11905771 TI - David Khayat: co-founder of The Charter of Paris Against Cancer (interview by Ezzie Hutchinson). PMID- 11905772 TI - The changing face of Russian oncology. PMID- 11905773 TI - Medicine in the media. PMID- 11905774 TI - Medical marijuana on trial. PMID- 11905775 TI - Fractal tumour contours question therapy strategies. PMID- 11905776 TI - Hopes to use the pill for ovarian cancer prevention. PMID- 11905777 TI - Indian group develops tools for oral cancer diagnosis. PMID- 11905778 TI - Oncology in Russia. PMID- 11905779 TI - Angiogenesis: tumour size is no guide to malignancy. PMID- 11905780 TI - Chemotherapy in the management of squamous-cell carcinoma of the head and neck. AB - Previously reserved for palliation, chemotherapy is now also a central component of several curative approaches to the management of patients with advanced-stage head and neck cancer. Here we review the results of both induction chemotherapy and chemoradiotherapy trials in patients with curable disease, and chemotherapy trials in patients with recurrent and metastatic disease, and we highlight current areas of investigation. Compared with traditional treatment modalities, chemotherapy given on induction schedules to patients with advanced laryngeal cancer allows greater organ preservation without compromise to survival; when given concomitantly with radiotherapy to patients with resectable or unresectable advanced disease, chemotherapy again improves survival. PMID- 11905782 TI - Late presentation of male breast cancer. PMID- 11905781 TI - Spot the differences: proteomics in cancer research. AB - Separation of thousands of cellular proteins by two-dimensional electrophoresis allows the detailed comparison of proteins from normal and diseased tissue. Mass spectrometry provides a fast and reliable way of characterising proteins of interest, particularly when the gene sequence of the source organism is known. The availability of the human genome sequence has opened up the possibility of identifying protein differences between normal and diseased tissue, thus providing the opportunity to search for tumour markers or for therapeutic targets. This new technology will give much-needed insight into the molecular mechanisms of tumour development and progression. PMID- 11905783 TI - Angiogenesis: pathological, prognostic, and growth-factor pathways and their link to trial design and anticancer drugs. AB - Angiogenesis is essential for tumour growth, invasion, and metastasis. Tumour blood vessels show many differences from normal vessels and are not genetically unstable, so they form a potentially key area for therapy of all types of cancer including leukaemias. Here we review current knowledge on the multiple pathways controlling tumour angiogenesis and assess which are the most clinically relevant. We also review the clinical evidence that angiogenesis affects the behaviour of cancer. Retrospective studies on intratumoral vascularisation suggest that it is an independent prognostic factor that merits prospective validation. Also, the presence of vascular endothelial growth factor in high concentrations in primary cancers is associated with poor prognosis. Key targets for drug development, current clinical trials, and the problems of developing drugs that do not have direct cytotoxic effects are reviewed. Recommendations are made on organising and monitoring antiangiogenic trials. PMID- 11905784 TI - New therapies, new directions: advances in the systemic treatment of metastatic colorectal cancer. AB - Colorectal cancer is the second leading cause of cancer death and it is clear that patients with metastatic disease have better quality of life and survival when given treatment. Despite four decades of experience of treating patients with fluorouracil, there remains considerable controversy about the optimum dose and scheduling, as well as biomodulation with leucovorin and methotrexate. However, irrespective of the dose and schedule, overall survival times are poor- about 1 year. Disappointingly, oral agents with similar mechanisms to fluorouracil do not improve survival rates in comparison with fluorouracil and leucovorin treatment. Irinotecan and oxaliplatin are newer agents that have improved the response rates for patients with metastatic disease when they are added to flurouracil and leucovorin. The combination of irinotecan, fluorouracil, and leucovorin has also improved overall survival. These are small advances in the fight against colorectal cancer, and further drug development is necessary. PMID- 11905785 TI - Pemetrexed disodium, a novel antifolate with multiple targets. AB - Pemetrexed disodium is a potent new antifolate which inhibits many folate dependent reactions that are essential for cell proliferation. Its primary target is thymidylate synthase but it also inhibits folate-dependent enzymes involved in purine synthesis. Cells that are resistant to antifolates are generally less resistant to pemetrexed, irrespective of the mechanism of resistance. Pemetrexed has shown good activity in preclinical models with human tumour cells and xenografts. In the majority of clinical trials of pemetrexed, the dose-limiting toxic effect is neutropenia; other side-effects are mostly gastrointestinal. Preclinical studies indicate that the toxic effects of pemetrexed can be reduced by dietary folate, resulting in an improved therapeutic index. Low folate status is also associated with higher levels of toxicity in patients. As a single agent pemetrexed has shown good activity against non-small-cell lung cancer, squamous cell carcinoma of head and neck, colon cancer, and breast cancer, and it appears to be particularly active in combination with cisplatin against non-small-cell lung cancer and mesothelioma. Phase II and III studies are underway. PMID- 11905786 TI - Circadian chronotherapy for human cancers. AB - Cell physiology is regulated by a 24-hour clock, consisting of interconnected molecular loops, involving at least nine genes. The cellular clock is coordinated by the suprachiasmatic nucleus, a hypothalamic pacemaker which also helps the organism to adjust to environmental cycles. This circadian organisation brings about predictable changes in the body's tolerance and tumour responsiveness to anticancer agents, and possibly also for cancer promotion or growth. The clinical relevance of the chronotherapy principle, ie treatment regimens based upon circadian rhythms, has been demonstrated in randomised, multicentre trials. Chronotherapeutic schedules have been used to document the safety and activity of oxaliplatin against metastatic colorectal cancer and have formed the basis for a new approach to the medicosurgical management of this disease, which achieved unprecedented long-term survival. The chronotherapy concept offers further promise for improving current cancer-treatment options, as well as for optimising the development of new anticancer or supportive agents. PMID- 11905787 TI - Larry Einhorn--President of ASCO, 2000-2001. American Society of Clinical Oncology (interview by Sue Silver). PMID- 11905788 TI - Humble pie. PMID- 11905789 TI - Elevation of CA15-3 caused by a benign tumour. PMID- 11905790 TI - Improving cancer care at the end of life. AB - Excellent care for cancer patients and their families should extend throughout the illness, and should include care provided at the end of life. Recent evidence, including a report from the Institute of Medicine, has emphasised that major reform is needed to improve relief of pain, other symptoms, and psychosocial care. This paper reviews the critical necessity for reform in end-of life care for the field of oncology and the major educational efforts required to ensure that oncology professionals can respond to this need. PMID- 11905791 TI - Tumour metastasis: is tissue an issue? AB - Single tumour cells, or multicellular aggregates, escape into blood and lymphatic vessels from a primary solid tumour as it progresses. However, the ability of such cells to develop into metastatic outgrowths is very limited, as shown by the poor prognostic power of the presence of circulating tumour cells in cancer patients. An explanation for this low efficiency may be a temporary absence of a suitable microenvironment once the tumour cells escape from their original tissue compartment. On the basis of histopathological observations, experimental studies, and the generally accepted poor prognosis of histopathologically confirmed intravascular tumour location, we propose that the structural and functional organisation of intravascular tumour cells as a tissue has a key role in providing the optimum microenvironment for sustained malignant dissemination. Such a tissue may be a fixed or a mobile intravascular microsatellite, or an intravascular micrometastasis, which locally develops into an overt in-transit metastasis. PMID- 11905792 TI - Paul Workman--at the cutting edge of drug discovery (interview by Dorothy Bonn). PMID- 11905793 TI - Prevention and the pleasure principle. PMID- 11905794 TI - Mixed messages about depleted uranium. PMID- 11905795 TI - Profit and science can co-exist. PMID- 11905797 TI - Breast cancer is a political issue. PMID- 11905796 TI - Uncommon cancers are also important. PMID- 11905798 TI - Radiation for prostate cancer. AB - The balance between tumour control and normal tissue damage with conventional radiotherapy is critical to outcome and morbidity in the treatment of localised prostate cancer. Recent technological advances have allowed a reduction in the amount of normal tissue included in target treatment volumes. This reduces morbidity and allows dose escalation, theoretically increasing the likelihood of tumour control. The methods used to achieve dose escalation are discussed and the available evidence for their safety and efficacy, relative to conventional treatment, is reviewed. Although there are no randomised studies to provide evidence of increased survival, the available evidence supports the hypothesis that dose escalation produces survival rates equivalent to surgical series and provides a realistic choice for patients. PMID- 11905799 TI - Targeting tumour vasculature: the development of combretastatin A4. AB - The requirement for neovascularisation to permit the development of solid tumours beyond a threshold size, has focused attention on the therapeutic potential of agents that prevent angiogenesis. The multistep nature of angiogenesis presents several targets for intervention, including the inhibition of the endothelial cell migration or proliferation normally associated with developing vessels. Compounds that damage established tumour vasculature are also of potential clinical use. We review the development of one such antivascular drug, combretastatin A4. This tubulin-binding agent was originally isolated from an African shrub, Combretum caffrum. The disodium combretastatin A4 phosphate prodrug is currently undergoing phase I clinical trials in the UK and USA. This review assesses the in vitro and in vivo data for combretastatin and the prodrug, and the preliminary data that have emerged from the phase I clinical trials. PMID- 11905800 TI - Synergistic interaction between Helicobacter pylori gastritis and diet in gastric cancer. AB - Infection with Helicobacter pylori increases the risk of gastric cancer. One possible mechanism is the higher likelihood of malignant transformation due to inflammatory responses in the epithelium. An alternative explanation is that these inflammatory responses induce chronic gastritis associated with decreased acidity in the stomach, which in turn increases the endogenous formation of carcinogenic N-nitroso compounds. Inflammatory responses seem to trigger two different causal pathways: one for the diffuse type of gastric cancer and the other for the intestinal type. The striking geographic variability in intestinal gastric cancer can be explained by the synergistic interaction between H. pylori infection and dietary factors, such as intake of salt and ascorbic acid. Screening and eradication of this organism, together with appropriate dietary modifications, offer promise in countries with a high prevalence of H. pylori infection and high risk of gastric cancer, but the safety of such interventions needs to be ensured. PMID- 11905801 TI - Telomerase: biology and phase I trials. AB - In normal somatic cells, the ends of chromosomes (the telomeres) shorten with each cell division. By contrast, in tumour cells, telomere length is maintained, generally through the reactivation of the reverse transcriptase enzyme, telomerase. At least three applications relating to telomeres and telomerase have been proposed: in cancer diagnosis and prognosis (especially through measurements of the catalytic component of telomerase, hTERT) and as a means of monitoring tumour response to therapy; as an aid to tissue engineering; and inhibition as a cancer therapeutic strategy. Mouse knockout, hTERT dominant negative, and antisense experiments suggest that telomerase inhibitors will confer anticancer activity, especially in tumours with short telomeres. Inhibitory strategies have focused on antisense molecules, inhibitors of reverse transcriptases, and small molecules able to interact with and stabilise four-stranded (G-quadruplex) structures formed by telomeres. Clinical trials involving telomerase inhibitors require careful consideration compared to those looking at conventional anticancer cytotoxic drugs. PMID- 11905802 TI - Chromosome segregation and cancer: cutting through the mystery. AB - Mitosis is the most dramatic--and potentially dangerous--event in the cell cycle, as sister chromatids are irreversibly segregated to daughter cells. Defects in the checkpoints that normally maintain the fidelity of this process can lead to chromosomal instability (CIN) and cancer. However, CIN--a driving force of tumorigenesis--could be the cancer cell's ultimate vulnerability. An important goal is to identify novel anticancer compounds that directly target the mitotic errors at the heart of CIN. PMID- 11905803 TI - Improving the efficacy of antibody-based cancer therapies. AB - A quarter of a century after their advent, monoclonal antibodies have become the most rapidly expanding class of pharmaceuticals for treating a wide variety of human diseases, including cancer. Although antibodies have yet to achieve the ultimate goal of curing cancer, many innovative approaches stand poised to improve the efficacy of antibody-based therapies. PMID- 11905804 TI - Cancer gene therapy: fringe or cutting edge? AB - Direct targeting of cancer cells with gene therapy has the potential to treat cancer on the basis of its molecular characteristics. But although laboratory results have been extremely encouraging, many practical obstacles need to be overcome before gene therapy can fulfil its goals in the clinic. These issues are not trivial, but seem less formidable than the challenge of killing cancers selectively and rationally--a challenge that has been successfully addressed. PMID- 11905805 TI - How melanoma cells evade trail-induced apoptosis. AB - At the doses used clinically, chemotherapy is believed to kill melanoma by a final common 'mitochondrial' pathway that leads to apoptosis. Similarly, several natural defence mechanisms kill melanoma by the same pathways. A corollary to the latter is that survival of melanoma in the host is due to the development of anti apoptotic mechanisms in melanoma cells. What are these mechanisms? And how might we bypass them to improve the treatment of melanoma? PMID- 11905806 TI - Microarray and histopathological analysis of tumours: the future and the past? AB - The excitement surrounding the development of DNA microarray analysis and proteomics has raised questions about the role of these techniques in clinical practice and patient management. But how did the traditional methods of cancer diagnosis and prognosis develop, and how can high-throughput techniques contribute? PMID- 11905807 TI - Two genetic hits (more or less) to cancer. AB - Most cancers have many chromosomal abnormalities, both in number and in structure, whereas some show only a single aberration. In the era before molecular biology, cancer researchers, studying both human and animal cancers, proposed that a small number of events was needed for carcinogenesis. Evidence from the recent molecular era also indicates that cancers can arise from small numbers of events that affect common cell birth and death processes. PMID- 11905808 TI - Actin' up: RhoB in cancer and apoptosis. AB - RhoB is a small GTPase that regulates actin organization and vesicle transport. It is required for signalling apoptosis in transformed cells that are exposed to farnesyltransferase inhibitors, DNA-damaging agents or taxol. Genetic analysis in mice indicates that RhoB is dispensable for normal cell physiology, but that it has a suppressor or negative modifier function in stress-associated processes, including cancer. PMID- 11905809 TI - Pharmacogenetics and cancer therapy. AB - Pharmacogenetics is the study of how genetic variations affect drug response. These variations can affect a patient's response to cancer drugs, for which there is usually a fine line between a dosage that has a therapeutic effect and one that produces toxicity. Gaining better insight into the genetic elements of both the patient and the tumour that affect drug efficacy will eventually allow for individualized dosage determination and fewer adverse effects. PMID- 11905810 TI - Knockout mice: a paradigm shift in modern immunology. AB - In the past decade, advances in genetic engineering and mouse knockout technology have transformed our understanding of the immune system. In particular, new perspectives on T-cell development, co-stimulation and activation have emerged from the study of single and multiple gene-knockout animals, as well as from conditional knockout and 'knock-in' mutants. Analysis of these animals has clarified important intracellular signalling pathways and has shed light on the regulatory mechanisms that govern normal immune responses and autoimmunity. PMID- 11905811 TI - How can immunology contribute to the control of tuberculosis? AB - Tuberculosis poses a significant threat to mankind. Multidrug-resistant strains are on the rise, and Mycobacterium tuberculosis infection is often associated with human immunodeficiency virus infection. Satisfactory control of tuberculosis can only be achieved using a highly efficacious vaccine. Tuberculosis is particularly challenging for the immune system. The intracellular location of the pathogen shields it from antibodies, and a variety of T-cell subpopulations must be activated to challenge the bacterium's resistance to antibacterial defence mechanisms. A clear understanding of the immune responses that control the pathogen will be important for achieving optimal immunity, and information provided by functional genome analysis of M. tuberculosis will be vital in the design of a future vaccine. PMID- 11905812 TI - Lymphostromal interactions in thymic development and function. AB - The generation of a peripheral T-cell pool is essential for normal immune system function. CD4+ and CD8+ T cells are produced most efficiently in the thymus, which provides a complexity of discrete cellular microenvironments. Specialized stromal cells, that make up such microenvironments, influence each stage in the maturation programme of immature T-cell precursors. Progress has recently been made in elucidating events that regulate the development of intrathymic microenvironments, as well as mechanisms of thymocyte differentiation. It is becoming increasingly clear that the generation and maintenance of thymic environments that are capable of supporting efficient T-cell development, requires complex interplay between lymphoid and stromal compartments of the thymus. PMID- 11905813 TI - Natural killer cells, viruses and cancer. AB - Natural killer cells are innate immune cells that control certain microbial infections and tumours. The function of natural killer cells is regulated by a balance between signals transmitted by activating receptors, which recognize ligands on tumours and virus-infected cells, and inhibitory receptors specific for major histocompatibility complex class I molecules. Here, we review the emerging evidence that natural killer cells have an important role in vivo in immune defence. PMID- 11905814 TI - Regulation of lymphocyte proliferation and death by FLIP. AB - Lymphocyte homeostasis is a balance between lymphocyte proliferation and lymphocyte death. Tight control of apoptosis is essential for immune function, because its altered regulation can result in cancer and autoimmunity. Signals from members of the tumour-necrosis-factor receptor (TNF-R) family, such as Fas and TNF-R1, activate the caspase cascade and result in lymphocyte death by apoptosis. Anti-apoptotic proteins, such as FLIP (also known as FLICE/caspase-8 inhibitory protein) have recently been identified. FLIP expression is tightly regulated in T cells and might be involved in the control of both T-cell activation and death. Abnormal expression of FLIP might have a role not only in autoimmune diseases, but also in tumour development and cardiovascular disorders. PMID- 11905815 TI - Man the barrier! Strategic defences in the intestinal mucosa. AB - Immunologists typically study the immune responses induced in the spleen or peripheral lymph nodes after parenteral immunization with antigen and poorly defined experimental adjuvants. However, most antigens enter the body through mucosal surfaces. It is now clear that the microenvironment in these mucosal barriers has a marked influence on the immune response that ultimately ensues. Nowhere is the microenvironment more influential than in the gut-associated lymphoid tissue (GALT). The GALT must constantly distinguish harmless antigens that are present in food or on commensal bacteria from pathogenic assault by microbes. It is perhaps not surprising, then, that the GALT contains more lymphocytes than all of the secondary lymphoid organs combined. PMID- 11905816 TI - The germless theory of allergic disease: revisiting the hygiene hypothesis. AB - Rising rates of allergic disease accompany the healthier benefits of a contemporary westernized lifestyle, such as low infant mortality. It is likely that these twinned phenomena are causally related. The hygiene hypothesis states that allergy and increased longevity are both consequences of reducing infectious stressors during early childhood for millennia. Mechanistic explanations for the hygiene hypothesis have typically invoked the T-helper-type 1/2 (T(H)1/T(H)2) model. Here, we discuss why we favour a broader 'counter-regulatory' model--one that might also explain the increasing incidence of autoimmune disease in westernized countries. PMID- 11905817 TI - Epstein-Barr virus: exploiting the immune system. AB - In vitro, Epstein-Barr virus (EBV) will infect any resting B cell, driving it out of the resting state to become an activated proliferating lymphoblast. Paradoxically, EBV persists in vivo in a quiescent state in resting memory B cells that circulate in the peripheral blood. How does the virus get there, and with such specificity for the memory compartment? An explanation comes from the idea that two genes encoded by the virus--LMP1 and LMP2A--allow EBV to exploit the normal pathways of B-cell differentiation so that the EBV-infected B blast can become a resting memory cell. PMID- 11905818 TI - Role of chemokines in the pathogenesis of asthma. AB - The prevalence of asthma has risen drastically in the last two decades, with a worldwide impact on health care systems. Although several factors contribute to the development of asthma, inflammation seems to be a common factor that leads to the most severe asthmatic responses. In the past decade, researchers have characterized a large group of chemotactic cytokines, also known as chemokines, which are implicated in asthmatic inflammation. These chemokines control and direct the migration and activation of various leukocyte populations. Targeting chemokines should lead to new ways of controlling the inflammatory asthmatic response. PMID- 11905819 TI - Towards a blood-stage vaccine for malaria: are we following all the leads? AB - Although the malaria parasite was discovered more than 120 years ago, it is only during the past 20 years, following the cloning of malaria genes, that we have been able to think rationally about vaccine design and development. Effective vaccines for malaria could interrupt the life cycle of the parasite at different stages in the human host or in the mosquito. The purpose of this review is to outline the challenges we face in developing a vaccine that will limit growth of the parasite during the stage within red blood cells--the stage responsible for all the symptoms and pathology of malaria. More than 15 vaccine trials have either been completed or are in progress, and many more are planned. Success in current trials could lead to a vaccine capable of saving more than 2 million lives per year. PMID- 11905820 TI - Cross-presentation in viral immunity and self-tolerance. AB - T lymphocytes recognize peptide antigens presented by class I and class II molecules encoded by the major histocompatibility complex (MHC). Classical antigen-presentation studies showed that MHC class I molecules present peptides derived from proteins synthesized within the cell, whereas MHC class II molecules present exogenous proteins captured from the environment. Emerging evidence indicates, however, that dendritic cells have a specialized capacity to process exogenous antigens into the MHC class I pathway. This function, known as cross presentation, provides the immune system with an important mechanism for generating immunity to viruses and tolerance to self. PMID- 11905821 TI - Toll-like receptors and innate immunity. AB - Toll-like receptors have a crucial role in the detection of microbial infection in mammals and insects. In mammals, these receptors have evolved to recognize conserved products unique to microbial metabolism. This specificity allows the Toll proteins to detect the presence of infection and to induce activation of inflammatory and antimicrobial innate immune responses. Recognition of microbial products by Toll-like receptors expressed on dendritic cells triggers functional maturation of dendritic cells and leads to initiation of antigen-specific adaptive immune responses. PMID- 11905822 TI - From T to B and back again: positive feedback in systemic autoimmune disease. AB - Systemic lupus erythematosus, a prototypical systemic autoimmune disease, is the result of a series of interactions within the immune system that ultimately lead to the loss of self-tolerance to nuclear autoantigens. Here, we present an integrated model that explains how self-tolerance is initially lost and how the loss of tolerance is then amplified and maintained as a chronic autoimmune state. Key to this model are the self-reinforcing interactions of T and B cells, which we suggest lead to perpetuation of autoimmunity as well as its spread to multiple autoantigen targets. PMID- 11905823 TI - Xenotransplantation and other means of organ replacement. AB - Exciting new technologies, such as cellular transplantation, organogenesis and xenotransplantation, are thought to be promising approaches for the treatment of human disease. The feasibility of applying these technologies, however, might be limited by biological and immunological hurdles. Here, we consider whether, and how, xenotransplantation and various other technologies might be applied in future efforts to replace or supplement the function of human organs and tissues. PMID- 11905824 TI - Vaccine safety--vaccine benefits: science and the public's perception. AB - The development of cowpox vaccination by Jenner led to the development of immunology as a scientific discipline. The subsequent eradication of smallpox and the remarkable effects of other vaccines are among the most important contributions of biomedical science to human health. Today, the need for new vaccines has never been greater. However, in developed countries, the public's fear of vaccine-preventable diseases has waned, and awareness of potential adverse effects has increased, which is threatening vaccine acceptance. To further the control of disease by vaccination, we must develop safe and effective new vaccines to combat infectious diseases, and address the public's concerns. PMID- 11905825 TI - Positive and negative regulation of T-cell activation by adaptor proteins. AB - Adaptor proteins, molecules that mediate intermolecular interactions, are now known to be as crucial for lymphocyte activation as are receptors and effectors. Extensive work from numerous laboratories has identified and characterized many of these adaptors, demonstrating their roles as both positive and negative regulators. Studies into the molecular basis for the actions of these molecules shows that they function in various ways, including: recruitment of positive or negative regulators into signalling networks, modulation of effector function by allosteric regulation of enzymatic activity, and by targeting other proteins for degradation. This review will focus on a number of adaptors that are important for lymphocyte function and emphasize the various ways in which these proteins carry out their essential roles. PMID- 11905826 TI - Autoreactivity by design: innate B and T lymphocytes. AB - Innate B and T lymphocytes are a subset of lymphocytes that express a restricted set of semi-invariant, germ-line-encoded, autoreactive antigen receptors. Although they have long been set apart from mainstream immunological thought, they now seem to represent a distinct immune-recognition strategy that targets conserved stress-induced self-structures, rather than variable foreign antigens. Innate lymphocytes regulate a range of infectious, tumour and autoimmune conditions. New studies have shed light on the principles and mechanisms that drive their unique development and function, and show their resemblance to another subset of innate lymphocytes, the natural killer cells. PMID- 11905827 TI - Emerging links between hypermutation of antibody genes and DNA polymerases. AB - Substantial antibody variability is created when nucleotide substitutions are introduced into immunoglobulin variable genes by a controlled process of hypermutation. Evidence points to a mechanism involving DNA repair events at sites of targeted breaks. In vertebrate cells, there are many recently identified DNA polymerases that inaccurately copy templates. Some of these are candidates for enzymes that introduce base changes during hypermutation. Recent research has focused on possible roles for DNA polymerases zeta (POLZ), eta (POLH), iota (POLI), and mu (POLM) in the process. PMID- 11905828 TI - The function of E- and Id proteins in lymphocyte development. AB - Helix-loop-helix proteins are essential factors for lymphocyte development and function. In particular, E-proteins are crucial for commitment of lymphoid progenitors to the B- and T-cell lineages. E-proteins are negatively regulated by the Id class of helix-loop-helix proteins. The Id proteins function as dominant negative inhibitors of E-proteins by inhibiting their ability to bind DNA. Here, we review the role of E-proteins and their Id protein antagonists in lymphocyte proliferation and developmental progression. In addition, we discuss how E protein activity and Id gene expression are regulated by T-cell receptor (TCR) and pre-TCR-mediated signalling. PMID- 11905829 TI - Cytokines and immunodeficiency diseases. AB - Severe combined immunodeficiency disease (SCID) refers to a spectrum of inherited immunodeficiencies that together represent the most severe forms of primary immunodeficiency in humans. Recent work has shown that many of these diseases, as well as other forms of immunodeficiency, result from defects in cytokine signalling pathways. Such defects can prevent normal development of lymphoid lineages and/or compromise cytokine signalling by these cells. These natural 'experiments' in human genetics have shown the non-redundant role for several cytokines or cytokine signalling molecules. Moreover, a comparison of the phenotypes of humans with SCID to analogous mouse-knockout models has shown not only expected similarities, but also unexpected differences in cytokine signalling between humans and mice. PMID- 11905830 TI - Strategies for designing and optimizing new generation vaccines. AB - Although the field of immunology developed in part from the early vaccine studies of Edward Jenner, Louis Pasteur and others, vaccine development had largely become the province of virologists and other microbiologists, because the model for classic vaccines was to isolate the pathogen and prepare a killed or attenuated pathogen vaccine. Only recently has vaccinology returned to the realm of immunology, because a new understanding of immune mechanisms has allowed translation of basic discoveries into vaccine strategies. PMID- 11905831 TI - T-cell regulation by CD28 and CTLA-4. AB - Activation of T lymphocytes is thought to require at least two signals, one delivered by the T-cell receptor complex after antigen recognition, and one provided on engagement of co-stimulatory receptors, such as CD28. Recent studies are providing clues as to the specific signalling roles of co-stimulatory receptors. Furthermore, superimposition of inhibitory signals, such as those delivered by cytotoxic T-lymphocyte antigen 4 (CTLA-4), leads to a complex network of positive and negative co-stimulatory signals, the integration of which modulates immune responses. PMID- 11905832 TI - Louis Pasteur's beer of revenge. AB - Although by the mid-nineteenth century evidence existed for an association between micoorganisms and disease, it was the combined efforts of Louis Pasteur and Robert Koch that created the germ theory of disease--the theory that specific microbes cause specific diseases. Surprisingly, the relationship between the two founders of microbiology and immunology was far from friendly. PMID- 11905834 TI - Transgenic analysis of thymocyte signal transduction. AB - This review examines the value of transgenic studies in mice for the genetic dissection of signal-transduction pathways relevant to thymus development. T-cell development in the thymus is controlled by an ordered sequence of differentiation and proliferation checkpoints that culminate in the production of correctly selected, non-autoreactive, peripheral T lymphocytes. Work in transgenic mice has been fundamental for the preparation of genetic maps of signal-transduction pathways that control T-cell development. This review discusses how tyrosine kinases, guanine-nucleotide-binding proteins and transcription factors converge to control T-cell differentiation and proliferation in the immune system. PMID- 11905833 TI - Transplantation tolerance from a historical perspective. AB - Although transplantation immunology as a distinctive field began with the development of experimental models that showed the feasibility of bone marrow transplantation, organ engraftment was accomplished first in humans, and was thought for many years to occur by drastically different mechanisms. Here, we present our view of the concepts of allograft acceptance and acquired tolerance from a historical perspective, and attempt to amalgamate them into simple and unifying rules that might guide improvements in clinical therapy. PMID- 11905835 TI - Modelling viral and immune system dynamics. AB - During the past 6 years, there have been substantial advances in our understanding of human immunodeficiency virus 1 and other viruses, such as hepatitis B virus and hepatitis C virus, that cause chronic infection. The use of mathematical modelling to interpret experimental results has made a significant contribution to this field. Mathematical modelling is also improving our understanding of T-cell dynamics and the quantitative events that underlie the immune response to pathogens. PMID- 11905836 TI - Cytokines and autoimmunity. AB - Cytokines have crucial functions in the development, differentiation and regulation of immune cells. As a result, dysregulation of cytokine production or action is thought to have a central role in the development of autoimmunity and autoimmune disease. Some cytokines, such as interleukin-2, tumour-necrosis factor and interferons--ostensibly, the 'bad guys' in terms of disease pathogenesis--are well known for the promotion of immune and inflammatory responses. However, these cytokines also have crucial immunosuppressive functions and so, paradoxically, can also be 'good guys'. The balance between the pro-inflammatory and immunosuppressive functions of these well-known cytokines and the implications for the pathogenesis of autoimmune disease is the focus of this review. PMID- 11905837 TI - Transforming growth factor-beta in T-cell biology. AB - Strict control of T-cell homeostasis is required to permit normal immune responses and prevent undesirable self-targeted responses. Transforming growth factor-beta (TGF-beta) has been shown to have an essential role in that regulation. Owing to its broad expression, and inhibitory effects on multiple cell types of the immune system, TGF-beta regulation is complex. Through advances in cell-specific targeting of TGF-beta signalling in vivo, the role of TGF-beta in T-cell regulation has become clearer. Recent in vitro studies provide a better understanding of how TGF-beta regulates T-cell homeostasis, through multiple mechanisms involving numerous cell types. PMID- 11905838 TI - T(H)1 and T(H)2 cells: a historical perspective. AB - Demonstration of the existence and functions of T helper (T(H))1 and T(H)2 cells has had an enormous impact on basic and applied immunology. T(H)1 and T(H)2 cells have a crucial role in balancing the immune response. In this article, I attempt to trace the historical events contributing to the development of the T(H)1/T(H)2 concept, the current state of play, and briefly discuss the future prospects for the field. PMID- 11905839 TI - A role for antigen in the maintenance of immunological memory. AB - The immune system has a memory that it exhibits in the enhanced and augmented responses the second time it meets an antigen. The memory is the result of a number of changes to the system brought about during the primary response. The most important of these changes is the formation of an expanded pool of antigen specific memory cells. One of the enduring questions in immunology is how these memory cells are maintained for such long periods. PMID- 11905840 TI - Expression of CD40 and CD40 ligand among cell populations within rheumatoid synovial compartment. AB - Augmented and prolonged expression of CD40 ligand (CD40L) was recently reported in lymphoid cells from human lupus patients, suggesting that CD40/CD40L pathway was involved in the pathogenesis of systemic autoimmune diseases. This study was thus designed to study the expression of CD40 and CD40L among cell populations within inflammatory joints of patients with rheumatoid arthritis (RA). The result showed that most B cells and monocytes in synovial fluids (SF) expressed CD40. Cultured synovial fibroblasts also stained positive for CD40. Regarding CD40L, we found that T cells as well as B cells could express CD40L. Compared with normal controls, RA patients had higher levels of CD40L+ T cells (8.71 +/- 17.69% vs 1.74 +/- 2.30%, P > 0.05) and CD40L+ B cells (7.71 +/- 7.64% vs 1.12 +/- 1.59% P < 0.05). After in vitro stimulation, T cells from RA patients had higher and longer CD40L expression than T cells from normal peripheral blood. For investigating the effect of CD40 expressed on synovial fibroblasts on TNF-alpha production in joint compartment, we used anti-CD40 antibody to bind CD40 on fibroblasts for one hour and then co-cultured with synovial fluid mononuclear cells. We found that the levels of TNF-alpha decreased in the presence of anti CD40 antibody. We concluded that there was an intrinsic hyperexpression of CD40L on lymphoid cells within rheumatoid joints, and synovial fibroblasts could contribute to articular inflammation through surface CD40 molecule. PMID- 11905841 TI - Functional defect of B lymphocytes in a patient with selective IgM deficiency associated with systemic lupus erythematosus. AB - Selective IgM deficiency (SIgMD) is a rare primary immunodeficiency disease, which is found in some patients with autoimmune diseases. The pathogenesis of SIgMD and the relationship of these diseases have remained unclear. The absence of secreted IgM was recently reported to accelerate the development of autoimmune diseases in lupus-prone lymphoproliferative (Ipr) mice. The reduction of secreted IgM production may relate with the progression of autoimmune diseases in human. We present a case of SIgMD associated with systemic lupus erythematosus (SLE), and examined the function and the IgM heavy chain gene of patient's lymphocytes. The number and the surface IgM expression of the patient's B cells were normal. In vitro stimulation of peripheral mononuclear cells by recombinant IL-2 and a B cell activator, Staphylococcus aureus Cowan strain I, could not overcome the reduction of IgM production, although the secreted form of IgM mRNA was detected. Sequence analysis of the IgM heavy chain gene and the IgM mRNA revealed no mutation or deletion. These findings suggested that SIgMD in this case was involved in the abnormality during B cell maturation. Further analysis is required to reveal the pathogenesis of SIgMD associated with SLE. PMID- 11905842 TI - Autoantibodies in primary Sjogren's syndrome: new insights into mechanisms of autoantibody diversification and disease pathogenesis. AB - Characterisation of autoantibodies and their target autoantigens in primary Sjogren's syndrome (SS) is an important entry point for studying this common systemic autoimmune disease. Diversification of anti-Ro/La responses is believed to occur by a process of determinant spreading following initiation of an autoimmune response to one component, possibly 52-kD Ro (Ro52). Recent evidence supports the ER-resident chaperone Grp78 as a potential candidate in the initiation of an autoimmune response against Ro52, by binding to a Grp78 binding motif in the COOH-terminal region of Ro52. The subsequent diversification of the anti-Ro/La response is influenced by distinct HLA class II alleles. Anti-salivary duct autoantibodies have been revisited and shown to be mimicked by cross reactive isoantibodies to AB blood group antigens. Identification of autoantibodies that act as antagonists at M3-muscarinic receptors represents an important advance. As well as contributing to the sicca symptoms, the functional effects of these autoantibodies may explain associated features of autonomic dysfunction in patients with SS. Anti-M3 receptor autoantibodies occur in both primary and secondary SS and allow Sjogren's syndrome to be viewed as a disorder of anti-receptor autoimmunity. PMID- 11905843 TI - I-Aq and I-Ap bind and present similar antigenic peptides despite differing in their ability to mediate susceptibility to autoimmune arthritis. AB - Susceptibility to collagen induced arthritis (CIA) in the murine model is linked to expression of the MHC class II alleles, I-Aq and I-Ar. We have examined the molecular basis for this MHC-linked susceptibility by studying the antigen presentation function of two class II molecules, I-Aq and I-Ap, that are closely related yet differ in mediating susceptibility to CIA. These class II molecules differ by only 4 amino acids, yet only mice expressing I-Aq develop CIA. Although the I-Ap molecule can bind the same immunodominant determinant from type II collagen as I-Aq, H-2p APC have difficulty generating I-Ap:CII peptide complexes when processing of CII is required. Immunization of H-2p mice with type II collagen (CII) generated only a weak T cell response when compared to H-2q mice, whereas immunization with the a CII peptide containing the dominant determinant induced a strong T cell response in both strains. In antigen presentation assays, H-2p APC were very inefficient in stimulating T cells when native CII was used as antigen, however they presented CII synthetic peptides with similar efficiency as H-2q APC. Processing and presentation of other antigens by H-2p APC was not affected. Using soluble class II binding assays, the affinity of I-Ap for the CII dominant peptide was 10 to 50 fold lower than I-Aq, however, this reduced affinity was not a general defect in I-Ap function. I-Aq and I-Ap had virtually identical affinities for binding other antigenic peptides. These data indicate that MHC-based susceptibility to autoimmunity may involve more than simple determinant selection and that the successful generation of an antigenic peptide by processing may be related to the overall affinity of the peptide for the MHC molecule. PMID- 11905844 TI - The role of natural killer cells in the induction of autoimmune gastritis. AB - A number of experimental models of organ-specific autoimmunity involve a period of peripheral T cell lymphopenia prior to disease onset. In particular, experimental autoimmune gastritis, induced in susceptible mouse strains by neonatal thymectomy, is a CD4+ T cell mediated autoimmune disease. We have previously demonstrated that this disease displays the hallmarks of a Th1 mediated DTH inflammatory response with an essential role for IFN-gamma very early in the pathogenesis of disease. Given the interplay between the innate and adaptive immune responses, a potential source of early IFN-gamma production in these lymphopenic mice is the innate immune response. Here we have assessed the contribution of innate immunity to the induction of experimental autoimmune gastritis, in particular, the role of natural killer (NK) cells in production of IFN-gamma. Analysis of NK cells and macrophages revealed no difference in either the number or activation status between euthymic and neonatally thymectomised mice. Furthermore, in vivo depletion of NK cells immediately after neonatal thymectomy of (BALB/cCrSlcxC57BL/6) F1 mice demonstrated no reduction in disease incidence compared to control groups of neonatally thymectomised mice. Therefore, we conclude that NK cells are not the primary source of IFN-gamma required for the pathogenesis of autoimmune gastritis following neonatal thymectomy but rather the small cohort of T cells in the periphery of lymphopenic mice are likely to be responsible for the IFN-gamma production. PMID- 11905845 TI - Viral infections and resulting inhibition of xenobiotic metabolizing enzymes as the possible cause of autoimmune hepatitis in a genetically predisposed person persistently exposed to harmful environmental factor(s). PMID- 11905846 TI - Characterisation of proton pump antibodies and stomach pathology in gastritis induced by neonatal immunisation without adjuvant. AB - It has previously been reported that neonatal BALB.D2 mice injected with native proton pump antigens without adjuvant develop an irreversible gastritis (Claeys et al, 1997). The ease of inititating gastritis in the neonate stands in contrast with the difficulty in initiating gastritis in adult mice that require repeated immunisation in adjuvant that is reversible following cessation of immunisation (Scarff et al, 1997). In view of these contrasting observations, we set out to ascertain whether we could confirm the observations in neonatal mice as well as further characterise the pathology and the autoantibody response. We found that neonatal gastritis-susceptible BALB/c mice (n=12), immunised with either pig or mouse gastric membranes in the absence of adjuvant, develop gastritis without circulating antibody to parietal cells detected by immunofluorescence, a hallmark of murine and human gastritis (Toh et al, 1997). However, mice immunized with pig gastric membranes (n=6) had circulating antibodies reactive by immunofluorescence to recombinant alpha and/or beta subunit of gastric H+/K+-ATPase expressed by insect cells (Sfalpha and Sfbeta). Four mice from this cohort with antibodies to Sfbeta also had reactivity to gastric H+/K+-ATPase by ELISA, and 3 immunoblotted the beta but not the alpha subunit of the ATPase. In the cohort of mice immunised with mouse gastric membranes (n=6), four produced antibodies reactive by immunofluorescence to Sfalpha, two of which were also reactive to Sfbeta and one developed antibodies detected by ELISA to gastric H+/K+-ATPase. However, no members of this cohort had antibodies reactive by immunoblotting to either the beta or alpha subunit of the ATPase. In all cases gastritic stomachs were characterised by areas deficient in ribosome-rich zymogenic cells and marked reductions in H+/K+-ATPase-positive parietal cells. Metaplasia detected by Maxwell stain, as clusters of mucus-producing cells throughout gastric units, were non-reactive to stomach mucin autoantibody suggesting the mucins comprise other and/or aberrant form(s). Compared to our previous observations in adult mice, our present data confirms that gastric autoimmunity is more readily induced in the neonate than the adult. Our data also affirms that while the neonatal immune system can mount a damaging inflammatory cellular immune response to gastric antigens, it develops an altered antibody response. PMID- 11905847 TI - Non-MHC driven exacerbation of experimental thyroiditis in the postpartum period. AB - Many human autoimmune diseases, including those of the thyroid gland, are affected by immune changes during pregnancy and the postpartum period. To investigate this influence, we have developed an animal model of pregnancy thyroiditis by using thyroglobulin (Tg)-induced experimental autoimmune thyroiditis (EAT). We now report a study of the post-partum period in mice with EAT. At 5 weeks postpartum, which was 9 weeks after the completion of a Tg immunization regime, the mean thyroiditis grade was significantly increased in the postpartum group from 0.23 to 0.43 (p<0.05) and the thyroiditis Index, which reflected both the frequency and severity of thyroiditis, was similarly increased compared to controls (29.0 vs 9.0). When Tg immunized CBA/J (H-2k) female mice were mated with BALB/c (H-2d) males, there was a similar increase in the severity of thyroiditis in the postpartum period as seen with CBA/J males suggesting that allogeneic factors were not able to further this postpartum exacerbation. Spleen cell IL-4 secretion was enhanced in the postpartum but only in the presence of thyroiditis indicating enhanced activity of Th2 immune responses. There were no differences in IFN-gamma secretion, titers of anti-Tg, CD8+ & CD4+ T cells and T cell chemokine receptor (CCR5, CCR3) expression between non-pregnant control mice with thyroiditis and postpartum thyroiditis. In summary, we found that the severity of EAT during the postpartum was significantly greater than in non pregnant control mice and was associated with enhanced Th2 immune responses. The allogenicity of the pregnancy had no influence on these findings. The lack of allogenic impact was in contrast to earlier observations in pregnancy itself where an exacerbation of thyroiditis was male strain-dependent and involved primarily Th1 responses. This indicated that the postpartum exacerbation of autoimmune thyroid disease was not a simple response to fetal antigens but secondary to unique postpartum factors. PMID- 11905848 TI - Preservation of enzyme activity and antigenicity after mutagenesis of the membrane anchoring domain of GAD65. AB - The smaller isoform of glutamic acid decarboxylase, GAD65, is an important autoantigen implicated in the pathogenesis of type 1 diabetes whereas the larger isoform, GAD67 appears to play no major role. The primary difference between the two isoforms resides in the N-terminal part of the molecule including the GAD65 membrane-anchoring domain. The aim of this study was to generate mutants of the membrane targeting domain spanning amino acids 24 to 31 of GAD65 to determine effects on enzyme activity and antibody recognition. Three GAD65 mutants were generated by substituting two, nine or eleven nucleotides coding for the membrane targeting with the corresponding bases of GAD67. SDS-PAGE and Western blotting wildtype (wt) and mutated GAD65 ascertained that they were of similar size and recognized GAD65-specific antibodies. No difference in enzymatic activity was found between the mutants and wt GAD65. GAD65 antibody positive sera from type 1 diabetes patients immunoprecipitated mutated GAD65 whether two, nine or eleven nucleotides were replaced. Mono-or polyclonal antibodies to the N-terminal region demonstrated that the mutated GAD65 with two or nine nucleotides replaced was immunoprecipitated markedly better than wt whereas no difference was detected using antibodies specific for the PLP-binding site in the middle part of GAD65 or the C-terminal region. Taken together, these data suggest that no major conformational changes have been introduced by mutating the membrane-anchoring domain of GAD65. PMID- 11905850 TI - The frequency of occurrence of autoantibodies against beta1-adrenoceptors and its clinical relevance in patients with hepatitis virus myocarditis. AB - The aim of this study was to examine the frequency of occurrence of autoantibodies against beta1-adrenoceptors in patients with hepatitis virus myocarditis (HVM) and its possible correlation with clinical characteristics. A total of 103 patients with viral myocarditis were divided into a positive group (HVM group, n=29) and a negative group (Non-HVM group, n=74) according to the laboratory findings regarding their type of hepatitis virus. The study parameters included UCG, ECG, biochemical findings and screening of autoantibodies against beta1-adrenoceptor. It was shown that the positive rate of the hepatitis virus was 28.16% (29/103) in patients with viral myocarditis. The severity of myocardial or liver injuries and the frequency of occurrence of autoantibodies against beta1-adrenoceptors in patients with viral myocarditis were more pronounced, before treatment, in the HVM group than in the Non-HVM group. The positive rates of the antibodies against the hepatitis virus and the autoantibodies against beta1-adrenoceptors were highly consistent in patients with HVM (p<0.05). In conclusion, the frequency of occurrence of the autoantibodies against beta1-adrenoceptors may be one important marker of HVM and, thus, possibly involved in the pathogenesis of the HVM. PMID- 11905849 TI - Complexity of human immune response profiles for CD4+ T cell epitopes from the diabetes autoantigen GAD65. AB - Complex protein antigens contain multiple potential T cell recognition epitopes, which are generated through a processing pathway involving partial antigen degradation via proteases, binding to MHC molecules, and display on the APC surface, followed by recognition via the T cell receptor. We have investigated recognition of the GAD65 protein, one of the well-characterized autoantigens in type I diabetes, among individuals carrying the HLA-DR4 haplotypes characteristic of susceptibility to IDDM. Using sets of 20-mer peptides spanning the GAD65 molecule, multiple immunostimulatory epitopes were identified, with diverse class II DR molecules functioning as the restriction element. The majority of T cell responses were restricted by DRB1 molecules; however, DRB4 restricted responses were also observed. Antigen-specific T cell clones and lines were derived from peripheral blood samples of pre-diabetic and IDDM patients and T cell recognition and response were measured. Highly variable proliferative and cytokine release profiles were observed, even among T cells specific for a single GAD65 epitope. PMID- 11905851 TI - Heterophile antibodies indicate progression of autoimmunity in human type 1 diabetes mellitus before clinical onset. AB - We previously reported serum cytokines in a group of long term non-progressors to Type 1 diabetes; this reactivity detected in ELISA is now identified as heterophile antibody in some sera. Here, we characterize heterophile antibody activity. A 14 kDa-polypeptide from heterophile antibody containing serum bound to an anti-IL-4 column, but IL-4 was not detected by Western blot or by MS/MS sequencing. However, in 2/13 heterophile antibody positive sera, T-cell growth was potentiated and was blocked by an anti-human immunoglobulin. To examine the relationship between low affinity heterophile antibody presence and disease progression, 1100 archived serum samples were analyzed with two pairs of antibodies from 443 diabetes-free first degree relatives of Type 1 diabetes mellitus patients for heterophile antibody; 95 individuals developed diabetes on follow-up. Twenty-two individuals, whose serum was heterophile antibody positive with the second pair of antibodies (but negative with the first pair of antibodies), had a significantly higher incidence of developing diabetes after five years. Thirty-seven individuals with heterophile antibody reactivity with the first pair of antibodies, regardless of reactivity with the second pair of antibodies, had a significantly lower incidence of developing diabetes. While we cannot exclude the presence of genuine cytokine in all sera, these data indicate the presence of distinct groups of heterophile antibodies in patients at high risk to develop diabetes. Thus, anti-Ig heterophilic antibodies with different immunochemical reactivities are linked to the progression of or protection from Type 1 diabetes autoimmunity. PMID- 11905852 TI - Methimazole interferes with the progression of experimental autoimmune myocarditis in rats. AB - In order to ascertain whether methimazole, a drug commonly used for the treatment of hyperthyroidism, interferes with the progression of autoimmune-mediated myocardial injury, we investigated the effect of methimazole on experimental autoimmune myocarditis (EAM) in rats. EAM was induced by immunization with porcine cardiac myosin. Methimazole administration markedly slowed the body weight growth in both normal and EAM rats, but did not induce morphologic change of cardiac tissue in normal rats. In EAM rats, macroscopic examination revealed discoloration of the cardiac surface, and histopathological examination by light microscopy showed extensive myocardial necrosis, infiltration by inflammatory cells and myocardial fibrosis. In the EAM rats treated with methimazole, the discolored areas on the cardiac surface were markedly diminished in size, and the myocardial necrosis, cellular infiltration and fibrosis were significantly less severe. To identify the mechanism responsible of this effect, we investigated the change of regulatory lymphocyte subsets in peripheral blood using an immunofluorescence technique with a flow cytometer. A decrease in the helper/suppressor T cell ratio as a result of the increased proportion of suppressor T cells and a decrease in the proportion of B cells were observed in normal rats after methimazole administration, and similar findings were made in the EAM rats treated with methimazole. These results indicate that methimazole interferes with the progression of EAM, and immunosuppression may, at least in part, be involved in the inhibitory effect of methimazole on EAM in rats. PMID- 11905853 TI - Echovirus 4 and type 1 diabetes mellitus. AB - AIMS/HYPOTHESIS: To determine the association between exposure to enteroviruses and Type 1 diabetes. METHODS: We measured neutralizing antibodies to the following enteroviruses: Coxsackievirus CA9, CB1, CB2, CB3, CB4, CB5, CB6, and Echovirus E4, E6, E9, E11 in the sera of (1) Type 1 diabetic patients at diagnosis (n = 33), (2) healthy offspring of parents with Type 1 diabetes without islet cell antibodies (ICA) (n = 43) and (3) normal controls (n = 57). All subjects were less than 20 years old. We performed the neutralization test determining the cytopathogenic effect on Vero cells. HLA DR serotyping was also performed in Group 2. RESULTS: Type 1 diabetic patients showed a higher frequency (21.2%, p < 0.01) of neutralizing antibodies to E4 in relation to controls (1.8%), although there were no differences comparing with offspring of Type 1 diabetic patients (20.9%). Healthy offspring carrying Type 1 diabetes HLA DR susceptibility genes were also exposed to E4 (15.0%). High frequencies of neutralizing antibodies to most enteroviruses were found in the control group. CONCLUSION: This study shows the association between Type 1 diabetes and the presence of neutralizing antibodies to Echovirus 4, suggesting the possible participation of this virus as an environmental trigger of this autoimmune disease. Interestingly, our population displays high frequencies of exposure to enterovirus (including CB4) although the incidence for Type 1 diabetes is low (2.9 per 100,000 inhabitants). PMID- 11905854 TI - Glomerular expression of Fas ligand and Bax mRNA in lupus nephritis. AB - The current studies were carried out to determine the expression of Fas ligand and Bax in kidneys from lupus nephritis as possible indicators of apoptosis. Twenty-four kidney biopsies from patients with lupus nephritis and 30 normal controls were studied for FasL and Bax gene expression by fluorescent in situ hybridization. Seventy percent of the lupus biopsies displayed FasL or Bax mRNAs. These genes were mainly expressed in biopsies with higher activity indices. In contrast, neither of these mediators was detected in normal glomeruli. These data suggest that FasL and Bax are up-regulated in lupus nephritis and may play a pathogenic role through apoptotic cascades. PMID- 11905855 TI - Rheumatoid arthritis and tumor necrosis factor alpha. PMID- 11905856 TI - Cost-effectiveness of laparoscopic vs open adrenalectomy: small savings in an expensive process. AB - BACKGROUND AND PURPOSE: Nowadays, laparoscopy has become the approach of choice for adrenalectomy, especially in cases of benign tumors <6 cm. The authors have studied, in a retrospective trial, two groups of patients who have undergone an adrenalectomy: 10 consecutive patients operated on by an open approach and 10 consecutive patients operated on by laparoscopy. METHODS: Laparoscopic adrenalectomies were performed via a transabdominal lateral approach, whereas open adrenalectomies were performed via an anterior transabdominal or posterior retroperitoneal approach. Clinical outcomes were recorded, and special attention was paid to the costs of both techniques, collecting economic data from the costs in outpatient visits, blood and urine tests, diagnostic imaging, hospital admissions prior to surgery, hospital admission for surgery, and surgical expenses. RESULTS: Operative time (110 vs 123 minutes), length of postoperative stay (3.7 vs 5.8 days), and time to oral intake (1 vs 2 days) were significantly lower in the laparoscopic group. From the economic point of view, however, there were no significant differences between laparoscopic and open groups (6,306 vs 7,581), and only surgical inhospital stay costs were significantly lower in the laparoscopic series (742 vs 1,191). All the costs generated by surgery (hospital admission for surgery plus surgical expenses) were smaller in the laparoscopic group but constituted only a small part of the general expenses for these patients. The more expensive part of the budget for every patient was the hospital admissions prior to surgery for diagnosis or preoperative treatment. CONCLUSION: Laparoscopy is a safe and comfortable approach for adrenalectomy and should be the technique of choice. From the economic point of view, laparoscopic adrenalectomy is cheaper than open adrenalectomy, but in all cases, surgical costs are only a minimal part of the budget, and the greater savings must come from the reduction in the presurgical diagnostic process. PMID- 11905857 TI - Low-pressure laparoscopy may ameliorate intracranial hypertension and renal hypoperfusion. AB - BACKGROUND: Increased abdominal pressure is associated with elevations in the intracranial pressure (ICP) and impaired renal function. These adverse effects are potentially important in clinical situations such as severe abdominal trauma and laparoscopic donor nephrectomy. It was hypothesized that the secondary elevation of ICP leads to release of vasoconstrictors, which may affect renal function by decreasing the renal blood flow (RBF). We investigated the effect of laparoscopy on ICP and renal blood flow in a porcine model. MATERIALS AND METHODS: The abdominal pressure of swine (N = 5; 20-25 kg) was gradually increased from baseline to 5, 15, and 25 mm Hg by insufflation of nitrogen into the abdominal cavity. The ICP was measured using a Camino monitor, and RBF was simultaneously measured using a Transonic Doppler probe placed on the renal artery. Results were analyzed using repeated measures ANOVA and the paired t test. RESULTS: No significant change from baseline was observed in ICP and RBF when the abdominal pressure was 5 mm Hg. However, both ICP and RBF were affected by increasing the abdominal pressure to 15 and 25 mm Hg (P = 0.035 and 0.04 for ICP and P = 0.074 and 0.034 for RBF, respectively). CONCLUSIONS: Low-pressure laparoscopy may reduce the adverse effects of pneumoperitoneum on ICP and RBF. It may be advisable to use low pressures in laparoscopic surgery, especially when changes in ICP or renal perfusion may have significant clinical implications. PMID- 11905858 TI - Laparoscopic cholecystectomy: an Indian experience of 1233 cases. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) is a well-established procedure for symptomatic cholelithiasis in India, but there are few data available regarding the procedure and its related complications. PATIENTS AND METHODS: This paper represents a retrospective review of 1233 patients who underwent LC at Government Medical College and Hospital, Chandigarh, India, over 4 years (1997-2000). The case files of all these patients were analyzed for patient particulars, intraoperative findings, reason for any open conversion, postoperative stay, and mortality. RESULTS: The overall conversion rate was 7.06% (87 patients). The commonest cause of conversion was a frozen Calot's triangle (52 patients), followed by injury to the common bile duct (8 patients). The average postoperative stay in successful LC was 1.32 days. The overall mortality rate was 0.16% (2 deaths). The quality of life after LC was good to excellent in more than 90% of patients. CONCLUSIONS: Despite multiple hands in training, the complication rates of LC are within acceptable limits. The overall conversion rate has risen because of the increase in elective conversions, but the incidence of complications has come down because of a "no hesitation" policy in converting. In spite of multiple operators, LC is the procedure of choice for symptomatic cholelithiasis at our hospital. PMID- 11905859 TI - The YAMA UroPatch sling for treatment of female stress urinary incontinence: a pilot study. AB - BACKGROUND AND PURPOSE: We report our initial experience using a new suburethral sling made from bovine pericardium for the treatment of urinary incontinence. To prevent rolling and curling of the sling, a unique anti-roll clip is incorporated into the UroPatch. In addition, the sling has a series of perforations that create evenly distributed tissue integration and avoid delayed seroma or hematoma formation, thereby reducing the risk of sling rejection, infection, and erosion. PATIENTS AND METHODS: Twenty-two female patients underwent suburethral sling procedures using the UroPatch. All patients demonstrated urethral hypermobility, intrinsic sphincteric deficiency, or both. Five patients had previous surgical treatment for urinary incontinence. RESULTS: All operations were completed successfully. No intraoperative or postoperative complications occurred. There was no evidence of local or systemic reaction to the UroPatch in any of the patients. With a mean follow-up of 20 months, sling rejection, erosion, or infection has not been demonstrated, and no sling required removal to date. Urinary incontinence was corrected in all but one case. CONCLUSION: The results of this pilot study suggest that the UroPatch is a promising alternative to current slings for the treatment of female urinary incontinence. PMID- 11905861 TI - Endoscopic band ligation: alternative treatment method in nonvariceal upper gastrointestinal hemorrhage. AB - BACKGROUND: In the treatment of acute upper gastrointestinal (GI) bleeding, endoscopic band ligation (EBL) may be performed for nonfibrotic superficial lesions. This method has recently gained popularity in the treatment of nonvariceal upper GI bleeding. PATIENTS AND METHODS: Band ligation was performed in 13 patients who were admitted with active upper GI bleeding between December 1998 and February 2001. The sources of the bleeding were Mallory-Weiss syndrome in four patients, Dieulafoy's ulcer in five patients, gastric ulcer near a gastrojejunostomy anastomosis in two patients, gastric angiodysplasia in one patient, and the primary repair site in the stomach of a gunshot wound in one. RESULTS: Bleeding from all lesions except one was managed successfully with EBL. The single failure was in bleeding from a gastric Dieulafoy's lesion. Injection sclerotherapy with 1:10,000 epinephrine solution and EBL was not successful. Rebleeding occurred twice in one patient, and the second rebleeding necessitated surgical treatment. CONCLUSION: Our results revealed that EBL is a very promising technique in acute nonvariceal upper GI bleeding. Its effectiveness and safety with few complications will allow this modality to be used more widely. PMID- 11905860 TI - Effect of laparoscopic failure on outcome of laparoscopic nissen fundoplication: independent review. AB - PURPOSE: This study was undertaken to determine whether conversion or early reoperation contributed significantly to the eventual outcome of laparoscopic Nissen fundoplication. PATIENTS AND METHODS: An independent surgeon, blinded to the operative events, administered two general and one system-specific quality of life tools to the first 100 consecutive patients booked for laparoscopic fundoplication in a community hospital, where the open conversion rate was 4 patients/surgeon and the early reoperation rate 1.5 patients/surgeon for the first 20 operations. Patients were also asked about need for medication, dysphagia, satisfaction (analog scale), and whether, if given it to do over, they would opt for surgery again. RESULTS: Of the original 100 patients, 40 were studied an average of 5 years after surgery: 26 completed laparoscopically and 14 with laparoscopic failure (13 conversions and 1 early reoperation). Patient characteristics in the two groups were similar, except that there were more older patients with more fixed intrathoracic hiatus hernias in the failure group. Among the parameters examined, no statistically significant differences could be detected between laparoscopic and converted patients. CONCLUSIONS: General and system-specific quality of life, digestive symptoms, need for medication, patient satisfaction, and willingness to have surgery over again are not altered by conversion or reoperation. Thus, surgeons who have adequate laparoscopic skills, experience with open fundoplication, and training in laparoscopic Nissen fundoplication should feel free to add it to their repertoires provided the likelihood of conversion and reoperation secondary to inexperience is understood by the patient. PMID- 11905862 TI - Technique for laparoscopy-assisted complicated peritoneal dialysis catheter placement. AB - Peritoneal dialysis is widely accepted for the chronic management of end-stage renal disease. Especially in patients suspected of having intra-abdominal adhesions, the application of laparoscopic surgical techniques has significantly changed our surgical approach to dialysis catheter placement. The blind placement of peritoneal dialysis catheters in this patient group can be both dangerous, because of the higher risk of bowel injuries, and unsuccessful, because of immediate catheter misplacement or entrapment. We describe a relatively simple step-by-step approach to laparoscopy-assisted peritoneal dialysis catheter placement with omentectomy in these more complicated cases. PMID- 11905863 TI - Video-assisted extracorporeal appendectomy. AB - BACKGROUND: Despite many randomized controlled trials, there are still several important doubts about laparoscopic appendectomy. Longer operative time and greater cost are the two major disadvantages of the laparoscopic technique. PATIENTS AND METHODS: In current techniques of laparoscopic appendectomy, the appendix is ligated and divided within the abdominal cavity. We have performed 423 laparoscopic appendectomies using an extracorporeal technique during the last 8 years. In this technique, the appendix is delivered to the surface through the cannula, and appendectomy is carried out extracorporally. RESULTS AND CONCLUSION: The extracorporeal technique of appendix removal can be performed as safely and efficiently as the open technique. We report our data on the series of 423 operations, in which we had the encouraging experience that the video-assisted extracorporeal technique is a practical and reasonable alternative to the traditional laparoscopic technique. PMID- 11905864 TI - Comparison of thermal spread after ureteral ligation with the Laparo-Sonic ultrasonic shears and the Ligasure system. AB - BACKGROUND AND PURPOSE: The extent of lateral spread of tissue injury is important in the success of a primary anastomosis. We compared the injury produced by the Laparo-sonic Coagulation Shears and the Ligasure system. MATERIALS AND METHODS: Ureters were harvested from domestic farm pigs and ligated with the Laparo-sonic (N = 13) or Ligasure (N = 9) system. The tissues were then fixed, stained, and examined by two pathologists without knowledge of the type of treatment. RESULTS: The mean length of thermal damage from the Ligasure was 2.11 mm (range 1.0-4.0 mm) whereas it was 1.92 (range 0.5-4.25 mm) from the Laparo sonic system. The difference is not statistically significant. CONCLUSION: Debridement of as much as 5 mm of each cut end produced by the Laparo-sonic or Ligasure system may be beneficial in reducing stricture and leak. PMID- 11905865 TI - An unusual case of Bouveret's syndrome. AB - A 69-year-old man presented to the emergency department with a 12-hour history of severe abdominal pain. His medical history was significant for a small-bowel obstruction that resolved with conservative therapy 4 months prior to admission. In the distant past, a Billroth II gastric resection was performed for ulcer disease. He was hypothermic, and laboratory studies showed elevated serum liver and pancreatic enzymes. A CT scan of the abdomen demonstrated fat stranding and a small amount of free air in the area of the pancreas. Gram-negative rods subsequently grew from blood cultures. A presumptive diagnosis of necrotising pancreatitis was made, and supportive care was instituted. Follow-up CT scan performed several days later demonstrated a large filling defect in the stomach. Endoscopy showed this defect to be a giant gallstone, and the diagnosis of Bouveret's syndrome was made. The patient underwent laparotomy. A duodenal perforation in the posterior aspect of the fourth portion was identified. The perforation had been caused by chronic impaction of the giant stone. The stone was removed through the perforation, and the perforation was closed in multiple layers. Drainage of the retroperitoneum was effected through large catheters placed through the flank. The presentation, diagnostic evaluation, treatment, and complications of this condition are discussed. PMID- 11905866 TI - Randomized comparison of conventional carbon dioxide insufflation and abdominal wall lifting for laparoscopic cholecystectomy. AB - BACKGROUND: Gasless laparoscopy using abdominal wall lifting (AWL) has been developed in an attempt to avoid the adverse effects of carbon dioxide pneumoperitoneum that may occur in conventional laparoscopy. However, lifting has been criticized for its poor operative space and surgical invasiveness. This study compared the AWL method with conventional CO2 pneumoperitoneum for laparoscopic cholecystectomy with respect to operation performance, postoperative course, and stress response. PATIENTS AND METHODS: During a 6-month period, 95 patients with symptomatic gallstones were randomly assigned to receive laparoscopic cholecystectomy with conventional CO2 pneumoperitoneum (CO2 group; N = 47) or the AWL method (AWL group; N = 48). Operative results and operative time were recorded. Cardiopulmonary functions were assessed, and arterial blood gases were analyzed during surgery. Urinary cortisol, vanillylmandelic acid, metanephrines, and nitrogen loss; serum complement 3, C-reactive protein, and interleukin-6; postoperative pain; and the presence of nausea and vomiting were assessed for 48 hours after surgery. Postoperative time to recovery of flatus, tolerance of a full oral diet, and full activity were also determined. RESULTS: Only three significant differences were found. First, intraoperative ventilatory function deteriorated significantly less in the AWL group. Second, arterial blood gas determinations and capnography showed a greater decrease in intraoperative arterial pH and compliance with CO2 retention and an increase in peak airway pressure in the CO2 group (P < 0.05), reflecting poorer ventilatory performance. Third, preparation time and total operating time were significantly greater with the AWL method (P < 0.05). CONCLUSIONS: Although AWL required a longer operation time, our results suggest that the technique may still have value in high-risk patients with cardiorespiratory diseases. PMID- 11905867 TI - Bilateral cryptorchidism with bilateral inguinal hernia and retrovesical mass in an infertile man: single-stage laparoscopic management. AB - A 30-year-old married man presented with the complaint of inability to procreate. Examination revealed bilateral nonpalpable testes and bilateral inguinal hernia. Ultrasonography of the abdomen could not locate the testis; instead, a hypoechoic 5 x 5-cm mass was found behind the bladder. A CT scan of the abdomen revealed the right testis near the right inguinal canal. The left testis could not be identified beside the soft tissue mass. The patient was taken for diagnostic as well as therapeutic laparoscopy. The testis on the right was found just proximal to the internal inguinal ring, and right orchidopexy was done. The left testis was small and rudimentary; hence, orchidectomy was done. Bilateral laparoscopic herniorrhaphy was carried out with polypropylene mesh by fixing it intracorporeally to the pubic bone, Cooper's ligament, inguinal ligament, and conjoint tendon. Subsequently, the retrovesical mass was excised and retrieved by dilating the umbilical port site. The operative time was 3.5 hours with minimal blood loss. The postoperative period was uneventful, and the patient was discharged after 24 hours. The histopathology examination of the retrovesical mass showed an extragonadal germ cell tumor compatible with seminoma. PMID- 11905868 TI - Spontaneous resolution of massive laparoscopy-associated pneumothorax: the case of the bulging diaphragm and review of the literature. AB - A massive left-side pneumothorax was identified intraoperatively on the basis of bulging left hemidiaphragm toward the end of an uncomplicated laparoscopic Nissen fundoplication. There were no changes in ventilatory or hemodynamic parameters. The pneumothorax was observed, and nearly total spontaneous resolution occurred in the recovery room within 1 hour. PMID- 11905869 TI - Literature watch. PMID- 11905870 TI - Impotence following radical prostatectomy: insight into etiology and prevention. 1982. PMID- 11905871 TI - Asymptomatic infections of the urinary tract. 1956. PMID- 11905872 TI - Differentiation of renal masses using A-mode ultrasound. 1971. PMID- 11905873 TI - Computed tomography of the kidney. 1977. PMID- 11905874 TI - Urinary tract imaging by nuclear magnetic resonance. 1982. PMID- 11905876 TI - Prognostic value of urodynamic testing in myelodysplastic patients. 1981. PMID- 11905875 TI - Diuretic radionuclide urography: a non-invasive method for evaluating nephroureteral dilatation. 1979. PMID- 11905877 TI - An operative technique for the correction of vesicoureteral reflux. 1958. PMID- 11905878 TI - Low pressure reflux: relation of vesicoureteral reflux to intravesical pressure. 1962. PMID- 11905879 TI - Vesical and ureteral damage from voiding dysfunction in boys without neurologic or obstructive disease. 1973. PMID- 11905880 TI - Transurethral urethrotomy under vision. 1978. PMID- 11905881 TI - A placebo-controlled double-blind study of the effect of phenoxybenzamine in benign prostatic obstruction. 1978. PMID- 11905882 TI - The effect of finasteride in men with benign prostatic hyperplasia. 1992. PMID- 11905883 TI - The correction of stress incontinence by simple vesicourethral suspension. 1949. PMID- 11905884 TI - A simplified surgical procedure for the correction of stress incontinence in women. 1959. PMID- 11905885 TI - Pubovaginal sling procedure for stress incontinence. 1978. PMID- 11905886 TI - Treatment of urinary incontinence by an implantable prosthetic urinary sphincter. 1974. PMID- 11905887 TI - Clean, intermittent self-catheterization in the treatment of urinary tract disease. 1972. PMID- 11905888 TI - Experience with new double J ureteral catheter stent. 1978. PMID- 11905889 TI - Bladder substitution after pelvic evisceration. 1950. PMID- 11905890 TI - Cutaneous vesicostomy. 1960. PMID- 11905891 TI - Urinary diversion via a continent ileal reservoir: clinical results in 12 patients. 1982. PMID- 11905892 TI - 25-year experience with replacement of the human bladder (Camey procedure). 1984. PMID- 11905893 TI - A one stage hypospadias repair. 1961. PMID- 11905894 TI - A one-stage hypospadias repair. 1970. PMID- 11905895 TI - Transverse preputial island flap technique for repair of severe hypospadias. 1980. PMID- 11905896 TI - Varicocele in subfertility. Results of treatment. 1955. PMID- 11905897 TI - Congenital curvature of the phallus: report of three cases with description of corrective operation. 1965. PMID- 11905898 TI - Small-Carrion penile prosthesis. New implant for management of impotence. 1975. PMID- 11905899 TI - Small-Carrion penile prosthesis: a report on 160 cases and review of the literature. 1978. PMID- 11905900 TI - Intracavernous injection of papaverine for erectile failure. 1982. PMID- 11905902 TI - Microsurgical vasovasostomy: a reliable vasectomy reversal. 1977. PMID- 11905901 TI - Oral sildenafil in the treatment of erectile dysfunction. 1998. PMID- 11905903 TI - Renovascular hypertension: the UCLA experience. 1979. PMID- 11905904 TI - Percutaneous trocar (needle) nephrostomy in hydronephrosis. PMID- 11905905 TI - Successful homotransplantation of the human kidney between identical twins. 1956. PMID- 11905906 TI - The use of polarized light in the analysis of calculi and in the study of crystals in tissue: a preliminary report on the method employed. 1941. PMID- 11905907 TI - The treatment of ureteral calculi. a report of 504 cases in which the Councill stone extractor and dilator was used. 1945. PMID- 11905908 TI - First clinical experience with extracorporeally induced destruction of kidney stones by shock waves. 1981. PMID- 11905909 TI - Nephrostolithotomy: percutaneous techniques for urinary calculus removal. 1982. PMID- 11905911 TI - Transurethral ureteroscopy in women: a ready addition to the urological armamentarium. 1978. PMID- 11905910 TI - Operative nephroscopy. 1972. PMID- 11905912 TI - Laparoscopic nephrectomy. 1991. PMID- 11905913 TI - The use of the smear of the urinary sediment in the diagnosis and management of neoplasm of the kidney and bladder. 1951. PMID- 11905914 TI - The results of radical nephrectomy for renal cell carcinoma. 1969. PMID- 11905915 TI - Conservative surgery for renal cell carcinoma: a single-center experience with 100 patients. 1989. PMID- 11905916 TI - Disease outcome in patients with low stage renal cell carcinoma treated with nephron sparing or radical surgery. 1996. PMID- 11905917 TI - Intracavitary Bacillus Calmette-Guerin in the treatment of superficial bladder tumors. 1976. PMID- 11905918 TI - Testicular tumors. A clinicopathological study. 1953. PMID- 11905919 TI - The thoracoabdominal approach for retroperitoneal gland dissection: its application to testis tumors. 1950. PMID- 11905920 TI - Cis-diamminedichloroplatinum, vinblastine, and bleomycin combination chemotherapy in disseminated testicular cancer. 1997. PMID- 11905921 TI - The value of serial measurement of both human chorionic gonadotropin and alpha fetoprotein for monitoring germinal cell tumors. 1976. PMID- 11905922 TI - The early diagnosis and radical cure of carcinoma of the prostate. Being a study of 40 cases and presentation of a radical operation which was carried out in four cases. 1905. PMID- 11905923 TI - Studies on prostatic cancer. I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. 1941. PMID- 11905924 TI - Prediction of prognosis for prostatic adenocarcinoma by combined histological grading and clinical staging. 1974. PMID- 11905925 TI - Purification of a human prostate specific antigen. 1979. PMID- 11905926 TI - Prostate cancer detection in a clinical urological practice by ultrasonography, digital rectal examination and prostate specific antigen. 1990. PMID- 11905927 TI - Retropubic prostatectomy: a new extravesical technique report on 20 cases. 1945. PMID- 11905929 TI - Transperineal 125iodine seed implantation in prostatic cancer guided by transrectal ultrasonography. 1983. PMID- 11905928 TI - Retropubic implantation of iodine 125 in the treatment of prostatic cancer. 1972. PMID- 11905930 TI - Definitive radiation therapy of carcinoma of the prostate. A report on 15 years of experience. 1973. PMID- 11905931 TI - Too wet to exercise? Leaking urine as a barrier to physical activity in women. AB - Leaking urine is frequently mentioned (anecdotally) by women as a barrier to physical activity. The aim of this paper was to use results from the Australian Longitudinal Study on Women's Health (ALSWH) to explore the prevalence of leaking urine in Australian women, and to ascertain whether leaking urine might be a barrier to participation for women. More than 41,000 women participated in the baseline surveys of the ALSWH in 1996. More than one third of the mid-age (45-50 years) and older (70-75) women and 13% of the young women (18-23) reported leaking urine. There was a cross-sectional association between leaking urine and physical activity, such that women with more frequent urinary leakage were also more likely to report low levels of physical activity. More than one thousand of those who reported leaking urine at baseline participated in a follow-up study in 1999. Of these, more than 40% of the mid-age women (who were aged 48-53 in 1999), and one in seven of the younger (21-26 years) and older (73-79 years) women reported leaking urine during sport or exercise. More than one third of the mid age women and more than one quarter of the older women, but only 7% of the younger women said they avoided sporting activities because of leaking urine. The data are highly suggestive that leaking urine may be a barrier to physical activity, especially among mid-age women. As current estimates suggest that fewer than half of all Australian women are adequately active for health benefit, health professionals could be more proactive in raising this issue with women and offering help through non-invasive strategies such as pelvic floor muscle exercises. PMID- 11905932 TI - Measurement of energy expenditure of daily tasks among mothers of young children. AB - There is currently some debate about whether the energy expenditure of domestic tasks is sufficient to confer health benefits. The aim of this study was therefore to measure the energy cost of five activities commonly undertaken by mothers of young children. Seven women with at least one child younger than five years of age spent 15 minutes in each of the following activities: sitting quietly, vacuum cleaning, washing windows, walking at moderate pace (approx 5km/hour), walking with a stroller and grocery shopping in a supermarket. Each of the six 'trials' was completed on the same day, in random order. A carefully calibrated portable gas analyser was used to measure oxygen uptake during each activity, and data were converted to units of energy expenditure (METS). Vacuum cleaning, washing windows and walking with and without a stroller were found to be 'moderate intensity activities' (3 to 6 METs), but supermarket shopping did not reach this criterion. The MET values for these activities were similar to those reported in the Compendium of Physical Activities (Ainsworth et al., 2000). However, the energy expenditures of walking, both with and without a stroller, were higher than those reported in the Compendium. The findings suggest that some of the tasks associated with domestic caring duties are conducted at an intensity which is sufficient to confer some health benefit. Such benefits will only accrue however if the daily duration of these activities is sufficient to meet current guidelines. PMID- 11905933 TI - A profile of Australian football injuries presenting to sports medicine clinics. AB - Most of the published data describing Australian football injuries is from hospital emergency departments and elite injury surveillance studies. There is a lack of good information about injuries to players at the lower levels of participation and those not severe enough to warrant hospital treatment. This study describes the profile of Australian football injuries that present to sports medicine clinics for treatment. New sports injury cases, presenting to five metropolitan Melbourne sports medicine clinics during a 12 month period in 1996-1997, were recorded through the Sports Medicine Injury Surveillance project. Both the patient and treating health professional provided personal and injury details. Australian football accounted for 29% of the 6479 recorded injury cases. The majority of injured players were male (99%) and from adult, community leagues (78%); the mean age was 23 years. Competition accounted for 78% of injuries and 72% of injured players presented for treatment to a sports physician/medical practitioner. Body contact accounted for half of all injuries and the most common injuries were medial ligament sprains of the knee (7%), lateral ligament sprains of the ankle (6%) and anterior cruciate ligament injuries (4%). In conclusion, sports medicine clinics treat a wide variety of football injuries and appear to be a good source of data about injuries to non-elite participants. PMID- 11905934 TI - Social physique anxiety and eating attitudes in female athletic and non-athletic groups. AB - One purpose of the present study was to compare Social Physique Anxiety (SPA) scores among four different female groups (physique-salient (PS) athletes aerobics competitors and divers, n = 63; weight-restricted athletes-rowers, n = 60; non-physique-salient (NPS) athletes--soccer, n = 75; and non-athlete students, n = 53). A second purpose of the study was to examine relations between SPA and disturbed eating attitudes among these four groups. Participants completed the Social Physique Anxiety Scale (SPAS; Hart, Leary, & Rejeski, 1989) and the Eating Attitude Test (EAT; Garner & Garfinkel, 1979). With respect to purpose one, an analysis of variance (ANOVA) with SPA (i.e., 9-item single factor model of the SPAS) as the dependent measure, failed to reach significance (F (3,184) = 1.61, p> .05). With respect to purpose two, significant relations were found between SPA and EAT scores for all four groups. Inferential comparison of the bivariate correlations (Fisher's Z transformations), however, showed no significant differences between groups. In addition, a moderated regression analysis was computed using EAT scores as the dependent measure. The variables were entered in the following order: SPA, followed by dummy vectors for each sport-type category, and then the interaction term between SPA and the dummy vectors. Results showed that there was no significant interaction effect. Thus, these analyses, taken together. suggest that the type of sport activity does not serve to moderate relations between SPA and disordered eating. Implications of the findings are discussed. PMID- 11905935 TI - Motor development and gender effects on stretch-shortening cycle performance. AB - A substantial part of human movement such as jumping, hopping, leaping and other bounding movements are improved by making a counter-movement. These activities are often described as stretch shortening cycle (SSC) movements. The aim of this study was to determine whether the SSC affects performance in vertical jump in children to the same extent as it does in adults. Comparisons were made between counter-movement jump (CMJ) and squat jump (SJ) performance in children and adults. The ratio of take-off velocity between jumps was used to measure performance of the SSC. Two groups of subjects comprising of 22 adults and 20 children performed three CMJ's and three SJ's from a force platform. Impulse, take-off velocity and power were obtained by numerical integration of the force n time traces. Performance was calculated from the velocity at take off. Both groups jumped significantly higher in the CMJ but there was a higher degree of variability in the performance of the children. The results indicated that children could utilise a SSC to enhance jumping performance. Variability in the take-off velocities in children, particularly in the SJ suggests the children performed this jump non-optimally. PMID- 11905936 TI - The energy cost of running on grass compared to soft dry beach sand. AB - This study compared the energy cost (EC) (J x kg(-1) x m(-1)) of running on grass and soft dry beach sand. Seven male and 5 female recreational runners performed steady state running trials on grass in shoes at 8, 11 and 14 km x h(-1). Steady state sand runs, both barefoot and in shoes, were also attempted at 8 km x h(-1) and approximately 11 km x h(-1). One additional female attempted the grass and sand runs at 8 km x h(-1) only. Net total EC was determined from net aerobic EC (steady state VO2, VCO2 and RER) and net anaerobic EC (net lactate accumulation). When comparing the surface effects (grass, sand bare foot and sand in shoes) of running at 8 km x h(-1) (133.3 m x min(-1)) in 9 subjects who most accurately maintained that speed (133.3 +/- 2.2 m x min(-1)), no differences (P>0.05) existed between the net aerobic, anaerobic and total EC of sand running barefoot or in shoes, but these measures were all significantly greater (P<0.05) than the corresponding values when running on grass. Similarly, when all running speed trials (n = 87) performed by all subjects (n = 13) for each surface condition were combined for analysis, the sand bare foot and sand in shoes values for net aerobic EC, net anaerobic EC and net total EC were significantly greater (P<0.001) than the grass running measures, but not significantly different (P>0.05) from each other. Expressed as ratios of sand to grass running EC coefficients, the sand running barefoot and sand in shoes running trials at 8 km x h(-1) revealed values of 1.6 and 1.5 for net aerobic EC, 3.7 and 2.7 for net anaerobic EC and 1.6 and 1.5 for net total EC respectively. For all running speeds combined, these coefficients were 1.5 and 1.4 for net aerobic EC, 2.5 and 2.3 for net anaerobic EC and 1.5 and 1.5 for net total EC for sand running barefoot and in shoes respectively. Sand running may provide a low impact, but high EC training stimulus. PMID- 11905937 TI - Influence of weight training exercise and modification of hormonal response on skeletal muscle growth. AB - To investigate the influence of carbohydrate (CHO) consumption on the acute hormonal response, and chronic adaptation to weight lifting exercise, two studies were conducted. Following a four-hour fast, seven young men (21.3 +/- 3.5 y) performed (on two occasions) a nine-station weight lifting protocol, completing 3 sets of 10 repetitions at 75% of 1RM (series 1). Randomly assigned, one session included the ingestion of a non-caloric placebo, and the other, a 6% CHO solution. For series 2, two groups of young men (21.3 +/- 1.5 y) participated in 12 weeks of progressive resistance weight training. Training for one group included the ingestion of a non-caloric placebo, and the other, a 6% CHO solution. In series 1, weight lifting exercise with CHO ingestion significantly (p < 0.05) elevated blood glucose and plasma insulin levels above baseline, as well as that occurring with the placebo. This resulted in a significant blunting of the cortisol response (7% with CHO compared to 99% with placebo). These findings indicate that CHO consumption during weight lifting exercise can modify the acute hormonal response to exercise. With series 2, CHO consumption continued to blunt the cortisol response to exercise during the twelve weeks of training. This is in contrast to significantly elevated cortisol levels observed for the placebo control group. Corresponding with the modified response patterns were differences in muscle growth. Weight training exercise with CHO ingestion resulted in significantly greater gains in both type I (19.1%) and type II (22.5%) muscle fibre area than weight training exercise alone. The difference in the cortisol response accounted for 74% of the variance (r = 0.8579, p = 0.006) of change in type I muscle fibre area, and 52.3% of the variance (r = 0.7231, p = 0.043) of change in type II muscle fibre area. These findings suggest that the modification of the cortisol response associated with CHO ingestion can positively impact the skeletal muscle hypertrophic adaptation to weigh training. PMID- 11905938 TI - Running economy is impaired following a single bout of resistance exercise. AB - The purpose of this study was to determine whether a low-volume high-intensity resistance training session influenced running economy during a subsequent aerobic treadmill run. Nine well trained distance runners (mean +/- SD; VO2max, 66.6 +/- 10.2 ml x kg(-1) x min(-1); weight, 65.8 +/- 10.2 kg; height, 173.4 +/- 7.8 cm; age 20 +/- 1.1 years) with resistance training experience performed treadmill running at two different speeds (0.56 m x sec(-1) and 0.20 m x sec(-1) below speed corresponding to lactate equilibrium) either rested or 1, 8 or 24 hours after a 50-minute whole body resistance training session. Running economy was assessed using open circuit spirometry while heart rate was recorded telemetrically. The contractile properties of the quadriceps femoris were also determined following each resistance training session and prior to each treadmill run using percutaneous electrical stimulation. Submaximal oxygen consumption was significantly increased one hour (2.6 +/- 2.3%, p= 0.007), and eight hours (1.6 +/- 2.5%, p= 0.032), but not 24 hours after resistance training. No significant differences were found in exercising heart rate, ventilation, respiratory exchange ratio, ratings of perceived exertion, or running mechanics. Peak twitch torque, time to peak torque, and half relaxation time of the quadriceps femoris were significantly reduced immediately following resistance training while peak twitch torque was also lower one hour following resistance training. Running economy following a resistance training session is impaired for up to 8 hours. This change was not paralleled by a concomitant change in exercising heart rate. The mechanism responsible for increased oxygen consumption following resistance training may be related to impairment of the force generating capacity of skeletal muscle, as there was a significant decrement in the contractile properties of the quadriceps femoris following resistance training. PMID- 11905939 TI - Visceral vascular injuries. AB - This article deals with injuries to the celiac trunk, superior and inferior mesenteric arterial injuires. Surgical approaches and physiological implications of interruption of the mesenteric arterial circulation are addressed in detail. Surgical techniques for the management of these injuries and the need for second look operations are also examined. PMID- 11905940 TI - Brachial and forearm vessel injuries. AB - Upper extremity vascular injuries are common in trauma. The mortality rate from these injuries is quite low; however, the morbidity rate is quite significant. Prompt diagnosis and treatment can reduce the amputation rate for these injuries to minimal. Furthermore, morbidity from late complications of chronic ischemia, restenosis, and cold intolerance can be decreased as well. Fasciotomy, although less frequently required than in lower extremity injuries, should be used in all cases of suspected compartment syndrome. PMID- 11905941 TI - Heroic procedures in vascular injury management: the role of extra-anatomic bypasses. AB - The insertion of an extra-anatomic bypass graft is an accepted operative technique in highly selected patients with atherosclerotic occlusive disease and contraindications to in situ grafting. In similar fashion, the technique should be considered in injured or septic patients with large soft tissue defects or wound infections overlying arterial repairs or involving native arteries. The combination of vigorous debridement of injured or infected soft tissue and insertion of an extra-anatomic bypass graft allows for appropriate care of the wound without concern for further injury to the now-displaced arterial repair. PMID- 11905942 TI - Vascular trauma and compartment syndromes. AB - Vascular injuries produce ischemia, and their repair produces reperfusion. Ischemia and reperfusion produce compartment syndrome. Although a local event, a compartment syndrome risks not only the affected extremity, but also the life of the patient. A high index of suspicion coupled with adequate knowledge of subtle clinical symptoms (and confirmed by intracompartmental pressure measurement) improve management of compartment syndrome, and this article discusses common pitfalls in its diagnosis and treatment. PMID- 11905943 TI - Complications of vascular injury management. AB - This article addresses failed arterial and venous repairs, thrombosed vessels and bypasses, postoperative pseudoaneurysms and arterio-venous fistulas. Management techniques for these complications are reviewed and morbidity and mortality rates provided. PMID- 11905944 TI - The European experience with vascular injuries. AB - The rich and diverse heritage of the management of vascular injuries in the 45 independent European countries prevents the authors from revealing a uniform picture of the European experience, but some trends are clearly emerging. In countries with a low incidence of penetrating trauma and increasing use of interventional vascular procedures, the proportion of iatrogenic vascular trauma exceeds 40% of all vascular injuries, whereas on other parts of the continent, armed conflicts are still a major cause of vascular trauma. National vascular registries, mostly in the Scandinavian countries, produce useful, nationwide data about vascular trauma and its management but suffer still from inadequate data collection. Despite a relatively low incidence of vascular trauma in most European countries, the results are satisfactory, probably in most cases because of active and early management by surgeons on call, whether with vascular training or not, treating all kinds of vascular surgical emergencies. In some countries, attempts at developing a trauma and emergency surgical specialty, including expertise in the management of vascular injuries, are on their way. PMID- 11905945 TI - Vascular trauma in Latin America: a regional survey. AB - As has been demonstrated, significant differences exist in demographics and the likelihood of accidents among Latin American countries; however, when figures were standardized, they showed a clear similarity in all the reviewed features of vascular trauma. A total of 66.4% of cases were managed solely on a clinical basis, with 78.9% of surgical procedures being performed within 6 hours of injury. Vascular repair was attempted in 84% of arterial injuries and 43% of venous injuries. Results are extremely good, with an 89% rate of success, especially considering that 63% of injuries were gunshot wounds and that the largest series, from Brazil, had a 21.3% rate of abdominopelvic injuries. The mortality rate amounted to 12.7%, but associated injuries, and particularly multiple trauma, account for 50.0% of the deaths. PMID- 11905946 TI - Vascular trauma in Colombia: experience of a level I trauma center in Medellin. AB - Trauma has become a major health problem in Colombia. The large number of trauma patients has made San Vicente de Paul Hospital of Medellin a major national referred trauma center. Under-reporting is a major problem in Colombia, as in other underdeveloped countries, because of the absence of automated information systems. Despite this and limited financial health resources, time to definitive treatment, morbidity, and mortality are similar to those of centers in developed countries. This article has covered the authors' experience with vascular injuries over a period of 5 years, representing 664 patients; the results were shown in this article. In addition, advances made in the development of new tools for the diagnosis of vascular trauma, such as helical CT angiography, were discussed. PMID- 11905947 TI - Iliac vessel injuries. AB - Trauma to the iliac vasculature continues to pose a significant challenge to management. In several large series, mortality for penetrating injuries is reported as approaching 40%. Uncontrollable hemorrhage originating from an anatomically inaccessible source and multiple associated injuries often contribute to this high mortality rate. This article discusses the current existing management strategies and the controversial role of PTFE in vascular reconstruction within a contaminated field. Concomitant injuries to the enteric viscera and genitourinary system are also addressed. Postoperative management including anticoagulation and the complications of liberal fasciotomy are mentioned. The evolving role of endovascular therapy as an adjunctive modality in the armamentarium of the trauma surgeon is also presented briefly. PMID- 11905949 TI - Vascular trauma in high-velocity gunshot wounds and shrapnel-blast injuries in Israel. AB - High-velocity gunshot and shrapnel-blast vascular injuries pose a great challenge and need to be approached in a systematic, multidisciplinary fashion. Early revascularization with temporary shunts, the use of autologous tissue, major venous reconstruction, a low threshold for fasciotomy, and reliable tissue coverage are the mainstays of management. PMID- 11905948 TI - Vascular injury in Australia. AB - Vascular injury poses a small but significant challenge in Australian trauma care. Opportunities such as better practice guidelines and minimum standards will allow surgeons to improve delivery of quality care to the next generation of vascular trauma victims. Training in the management of vascular trauma surgery with integration of vascular and general surgery in trauma care should optimize outcomes. The authors' vision is that all vascular and general surgery trainees would eventually undertake the Definitive Surgical Trauma Care Course and improve vascular trauma outcomes and reduce mortality. PMID- 11905950 TI - New approaches to trauma management using severity of illness and outcome prediction based on noninvasive hemodynamic monitoring. AB - The mathematical model satisfactorily predicted outcome in acute emergencies based on noninvasively monitored flow, pressure, pulse oximetry, tissue perfusion values, and their cumulative deficits. A decision support system provided information on the relative effectiveness of various therapeutic modalities based on the responses of patients with very similar states. The concept that hypovolemia and oxygen debt is an early primary problem that plays an important role in low flow and poor tissue perfusion states is supported by direct observation of massive hemorrhage, estimated blood loss of hemoperitoneum and hemothorax at the time of surgery, and prior studies in the literature that documented blood volume deficits in posttraumatic and postoperative patients who subsequently developed organ failures and death. PMID- 11905951 TI - Femoral vessel injuries. AB - Early diagnosis, expeditious vascular repair, and aggressive management of complications have resulted in an amputation rate of less than 9%. Repair rather than ligation of an associated femoral vein injury is commonly practiced by experienced trauma surgeons. In most circumstances, a reversed autogenous saphenous vein graft from the contralateral extremity is the conduit of choice; however, if a saphenous vein cannot be used because of size discrepancies, multiple associated trauma, or extensive contamination, polytetrafluoroethylene can be used with good results. If vein ligation is performed, early fasciotomy is indicated for close and meticulous monitoring of the compartmental pressures. Clearly, the most crucial components for a successful outcome are a thorough evaluation, early operation, and a flawless vascular repair. PMID- 11905952 TI - Popliteal vascular injuries. AB - Popliteal vascular injury remains one of the most difficult diagnostic and therapeutic challenges for trauma surgeons. Only with strict attention to rapid diagnosis; early surgical treatment with meticulous technical skill; and aggressive use of various adjunctive measures, such as completion arteriography, anticoagulation, fasciotomy, and proper prioritization of management of multiple injuries, can limb salvage be optimized. PMID- 11905953 TI - Shank vessel injuries. AB - The management of lower extremity vascular injuries has undergone dramatic changes over the last century. With the optimal management of femoral and popliteal injuries established, controversy still exists with respect to management of vascular injuries below the popliteal fossa, in the shank arterial vessels. These injuries are uncommon, often limb threatening, and usually require complex management decisions. Incidence of shank vessel injuries, imaging studies required for accurate and expedient diagnosis, determinants influencing the decision for repair or amputation, and details of techniques in surgical intervention are discussed. PMID- 11905954 TI - Isoflavones, substances with multi-biological and clinical properties. AB - Isoflavones, rich in soybean, are currently receiving much attention because of their potential role in preventing and treating cancer and other human chronic diseases. The present review provides an overview of the recent results in this research field. Data from epidemiological reports and laboratories have shown that isoflavones have multi-biological and pharmacological effects in animals and humans. These include estrogenic and antiestrogenic effects, cell signalling conduction, as well as cell growth and death. Based on these properties, soy protein and isoflavones have been associated with reduced incidences of breast and prostate cancers, cardiovascular diseases or osteoporosis, and exhibit some other favorable effects. The mechanism through which isoflavones may exert the above-mentioned functions are not only based on the estrogenic properties of isoflavones, but also on their role as protein tyrosine kinase inhibitors, as regulators of gene transcription, modulators of transcription factors, antioxidants, as well as by altering some enzyme activities. However, to draw a clear conclusion regarding the clinical use of isoflavones further investigation would be required, although only a few effects of short- or long-term use of soy proteins are known in humans. PMID- 11905955 TI - Effects of maternal chronic alcohol administration in the rat: lactation performance and pup's growth. AB - A fostering/crossfostering analysis of the effects of maternal ethanol exposure on lactation performance and offspring growth was performed. Wistar rats were kept under one of the three experimental nutritional treatments: alcohol-treated (EG), pair-fed-treated (PFG) (as a nutritional control of alcohol-associated malnutrition), and control or normal diet (CG). Rats from the EG group were accustomed to increased amounts of ethanol (5% during the first week to 20% in the fourth week). The 20% ethanol level was maintained throughout three additional weeks and during gestational and lactational period. Daily food intake, fluid consumption, body weight and gestational parameters were studied in control (CG), pair-fed (PFG) and ethanol dams (EG). At birth, half the litters were fostered to other dams of the same treatment (GLG) and half were cross fostered to dams of the opposite treatment (GG, LG). No cross-fostering analyses were performed on the pair-fed group. Offspring body weight was controlled throughout lactation. Liver, kidney and spleen weights as well as milk consumption were also studied at the end of lactation period. In dams, a significant reduction of body weight was described throughout the suckling period. No ethanol detrimental effects were observed on body weight at birth, but in spite of a normal birth weight, alcohol during lactation was responsible for a growth deficit. Milk consumption was significantly reduced in offspring exposed to ethanol during gestation and/or lactation. Curiously, prenatal alcohol exposure affects adversely the suckling behaviour in pups at the time of weaning. In our study, alcohol treatment and malnutrition affects liver and spleen weights. However, malnutrition decreases spleen weights more than alcohol treatment. In the case of the kidney weights the alcohol decreases kidney weight more than malnutrition. Collectively, the data from the present study show similar effects following pre/postnatal and postnatal alcohol exposure. The findings suggest that chronic alcohol administration during gestation and/or lactation adversely affects pup growth at weaning as indicated by its effect on milk consumption, pup and organ weight. PMID- 11905956 TI - Effect of 24-h food deprivation on lipoprotein composition and oleoyl-estrone content of lean and obese Zucker rats. AB - BACKGROUND: Food deprivation induces the mobilization of fat reserves and, consequently, the transport of lipids in plasma. Zucker obese rats are grossly hyperlipidemic and do not use lipids as an efficient energy substrate. They also have lower circulating levels of acyl-estrone than expected because of their large fat stores. AIM OF THE STUDY: To measure the effect of 24 h food deprivation on hyperlipidemia and acyl-estrone distribution in plasma in Zucker obese rats. METHODS: The plasma lipoprotein distribution and composition of Zucker lean (Fa/?) and obese (fa/fa) rats was determined after 24 hours of food deprivation. Lipid classes: phospholipid, free and esterified cholesterol and triacylglycerols, and protein and total (mainly acyl-) estrone were also measured in total plasma and lipoprotein fractions. RESULTS: Food-deprived rats showed lower triacylglycerol levels than fed rats, but obese rats maintained high lipid levels, mainly in the VLDL fraction. The ratio of total plasma free-to-esterified cholesterol was lower in fed lean rats (0.29) than in the obese (0.61); the situation improved slightly after 24-h starvation, since the corresponding ratios were 0.30 and 0.41. Acyl estrone levels changed little with 24-h food deprivation. The chylomicra + VLDL total estrone compartment was essentially unchanged in lean and obese fed and starved groups, but the HDL pool decreased with food deprivation in the obese. CONCLUSION: Short-term starvation helped to enhance the differences between lean and obese Zucker rats in the handling of lipoprotein lipids, the latter showing a marked impairment in their ability to dispose of circulating lipids. The different pace of plasma lipid utilization may compound the problems of cholesterol transfer, partly explaining the dyslipemia that characterizes this animal model of obesity. The differences in acyl-estrone distribution also indicate that fat mass is preserved more effectively in obese rats even after food deprivation. PMID- 11905957 TI - Impact of diets containing corn oil or olive/sunflower oil mixture on the human plasma and lipoprotein lipid metabolism. AB - BACKGROUND: The effects of monounsaturated fatty acids (MUFA) rich diets compared to those that are rich in polyunsaturated fatty acids (PUFA) as well as the effects of an intake of single oils compared to oil mixtures are controversially discussed and results are contradictory. AIM OF THIS STUDY: To evaluate the effects of a plant oil-mixture (olive/sunflower oil; saturated/monounsaturated/polyunsaturated (S/M/P) = 14:69:17) high in oleic acid but also showing a moderate content of polyunsaturated fatty acids (PUFA) in comparison with a single, PUFA rich corn oil (S/M/P = 13:33:54) used in a normal, balanced diet on human plasma and lipoprotein metabolism. METHODS: The double blind designed study comprised 28 healthy, non-smoking young men aged between 19 and 31 years. After two weeks of adjustment (mixed, balanced diet: 11.6 MJ average, average fat intake approximately 105 g/d), the design included a two week test period in which a diet with 80 g corn oil/d vs a mixture of 68 g olive- and 12 g sunflower oil/d (total 80 g) as the main fat source was given, followed by a crossover after two weeks. Compliance and ingestion of diets were monitored by assessing the fatty acid pattern in LDL and by determination of alpha- and gamma-tocopherol in plasma and LDL. Results Diets were well incorporated due to the significant changes in plasma- and LDL-tocopherol levels and the significant different average ratio of oleic acid to linoleic acid in LDL. The PUFA-rich corn oil diet was able to reduce low density lipoprotein (LDL) cholesterol from adjustment to T2 significantly (p < 0.01), which was also confirmed by a trend after cross over (p=0.15). Total cholesterol (only after cross over at T3), total triglycerides (TG) and very low density lipoprotein (VLDL)-TG were significantly lower at T2 after the corn oil diet than after the mixed oil diet. Total high density lipoproteins (HDL) and HDL cholesterol remained unchanged by both diets. CONCLUSIONS: The results show that during the intervention of two weeks for each diet and the following cross over the corn oil diet had more influence on lipoprotein metabolism than the MUFA-rich diet. The hypocholesteremic effect of the PUFA-rich diet must also be connected with the high amount of unsaponifiable substances, mainly phytosterols in the corn oil. PMID- 11905958 TI - A two-dimensional map and database of soluble nuclear proteins from HepG2 cells as reference for identification of nutrient-regulated transcription factors. AB - BACKGROUND: Eukaryotic cells of higher organisms are able to regulate gene transcription in response to changes in the supply of nutrients. In hepatocytes, extracellular glucose levels affect transcription of genes that encode enzymes engaged in glycolysis, gluconeogenesis and lipogenesis. While glucose response elements have been located within a few model gene promoters, the identity of glucose-sensing transcription factors and the mechanisms of their activation remain to be elucidated. AIM OF THE STUDY: We intended to establish a two dimensional map of nuclear proteins as a reference for identification of nutrient regulated transcription factors. METHODS: Human hepatoma HepG2 cells were used for the preparation of nuclear extracts. 150-200 microg of the protein mixture were analyzed by 2-dimensional gel electrophoresis (2-DE) and silver-stained protein spots were identified by MALDI-TOF mass spectrometry. RESULTS: Nuclear extracts capable of transcriptional initiation and elongation and containing low amounts of cytoplasmic contaminations were prepared. 543 spots between 17 and 100 kDa and pI 3.7 and 8.8 have been resolved. From these, 65 spots were analyzed by MALDI-TOF mass spectrometry and 53 spots were identified as known proteins of which six represented transcription factors. Regulation by glucose was shown for the activator protein-1 component cJun. Since cJun was not visible on the silver stained 2-DE gel, western blotting of 1-DE gels and immunological detection had to be used in this case. The data were used to construct an online database. CONCLUSIONS: A 2-DE map and database of soluble nuclear proteins is presented. The identification of several transcription factors was possible on the silver stained gels. However, further fractionation of the nuclear extracts will facilitate the detection of larger numbers of transcriptional regulators. The database and 2-DE map shown here may provide a useful reference for the identification of transcription factors from liver nuclei that are activated by different stimuli, e. g., nutrients. PMID- 11905960 TI - Telomeres, telomerase, and stability of the plant genome. AB - Telomeres, the complex nucleoprotein structures at the ends of linear eukaryotic chromosomes, along with telomerase, the enzyme that synthesizes telomeric DNA, are required to maintain a stable genome. Together, the enzyme and substrate perform this essential service by protecting chromosomes from exonucleolytic degradation and end-to-end fusions and by compensating for the inability of conventional DNA replication machinery to completely duplicate the ends of linear chromosomes. Telomeres are also important for chromosome organization within the nucleus, especially during mitosis and meiosis. The contributions of telomeres and telomerases to plant genome stability have been confirmed by analysis of Arabidopsis mutants that lack telomerase activity. These mutants have unstable genomes, but manage to survive up to ten generations with increasingly shortened telomeres and cytogenetic abnormalities. Comparisons between telomerase-deficient Arabidopsis and telomerase-deficient mice reveal distinct differences in the consequences of massive genome damage, probably reflecting the greater developmental and genomic plasticity of plants. PMID- 11905959 TI - Effects of alpha-tocopherol, beta-carotene and ascorbic acid on oxidative, hormonal and enzymatic exercise stress markers in habitual training activity of professional basketball players. AB - BACKGROUND: Intense physical exercise has been associated with an increase of free radical production. When the body's natural defense systems against free radicals are overwhelmed, oxidative stress increases. AIM OF THE STUDY: This study examined the effects of a vitamin antioxidant supplement, (composed of 600 mg alpha-tocopherol, 1000 mg ascorbic acid and 32 mg beta-carotene) on oxidative, hormonal, and enzymatic exercise stress markers during habitual training activity over 35 days. METHODS: The plasma concentrations of ascorbic acid, alpha tocopherol, beta-carotene, testosterone, cortisol and lipid peroxides and the serum activities of lactate dehydrogenase and creatine kinase were measured at four time points: pre-supplementation (PS), pre-training (PT), after training (AT) and 24 h after training (24h-AT) in 13 professional basketball players of the first Spanish Basketball League (ACB). RESULTS: Antioxidant supplementation led to a significant increase of alpha-tocopherol and beta-carotene from PS to PT. Plasma lipid peroxides decreased about 27.7% after 35 days of antioxidant treatment. A significant decrease of lactate dehydrogenase serum activity was observed during the 24 h recuperation time. During this time the anabolic/catabolic balance increased about 29.8% in the antioxidant supplemented group, although this increase did not reach statistical significance. CONCLUSIONS: The results of the present study suggest that supplementation with alpha-tocopherol, beta-carotene and ascorbic acid might partially account for the hormonal and enzymatic stress marker profile observed during habitual training activity of professional basketball players. PMID- 11905961 TI - Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara. AB - The gene for a thermal hysteresis (antifreeze) protein (sthp-64) from the bittersweet nightshade, Solanum dulcamara, was cloned and characterized. An expression cDNA library prepared from November S. dulcamara was screened using a polyclonal antibody generated against a previously purified 67 kDa thermal hysteresis protein, and positive clones were identified and sequenced. The full length thermal hysteresis protein gene was cloned into an Escherichia coli expression vector and expressed as a fusion protein. The putative thermal hysteresis protein (STHP-64) contains two conserved regions 56 and 57 amino acids in length which have the C-X4-C-X22-23-H-X1-H zinc finger motif which is present in WRKY proteins, a family of transcription factors which play a role in regulating expression of pathogenesis-related proteins in plants. Additional features of transcription factors, such as an acidic domain between the two zinc fingers and a glutamine-rich region upstream of the first zinc-finger are also present in STHP-64. A DNA binding assay showed that the expressed STHP-64 fusion protein has specific DNA-binding ability. A unique feature of STHP-64 is that the C-terminus contains 10 consecutive 13-mer repeats. Such repeats are a common feature of animal antifreeze proteins. The expressed STHP-64 fusion protein had low levels of thermal hysteresis activity, but this activity was considerably increased by addition of citrate, which is known as an enhancer of certain insect antifreeze proteins. Northern blots demonstrated that the STHP-64 transcript was not present in leaves until November and December, suggesting that cold acclimation induces STHP-64 production. PMID- 11905962 TI - Expression of the ACC synthase and ACC oxidase coding genes after self pollination and incongruous pollination of tobacco pistils. AB - In tobacco, as in other species, ethylene is produced in response to pollination. Although tobacco is a self-compatible species, it displays unilateral incongruity with other Nicotiana plants. Incongruous pollination also results in ethylene production, but this production differs depending on the pollen used and is related to the extent to which pollen tubes grow in the tobacco style. In the investigation reported here we followed the expression of the ACC synthase- and ACC oxidase-coding genes upon pollination of tobacco pistils and compared self pollination with incongruous pollination. The pattern of expression of these genes also correlated with pollen-tube growth, although wounding alone cannot explain the results obtained. We also examined the expression of these genes upon pollination of immature tobacco pistils, in which different pollen tubes grew indistinctly inside the tobacco style and reached the ovary at the same rate. In this situation no significant differences in gene expression could be observed between the different pollinations. Ethephon, a substance that produces ethylene, could, in some cases, minimize the arrest of incongruous pollen tubes inside the style. PMID- 11905963 TI - Molecular and biochemical characterization of an Arabidopsis thaliana arogenate dehydrogenase with two highly similar and active protein domains. AB - The present study reports the first molecular characterization of a plant arogenate dehydrogenase, the enzyme that catalyses the transformation of arogenate into tyrosine. The structure of the Arabidopsis thaliana tyrA gene is very peculiar since it encodes two highly similar, and putatively active, protein domains. PCR analyses confirmed the existence of a transcript encoding the two protein domains. The complete coding sequence and sequences corresponding to the two separate domains were independently cloned into Escherichia coli mutant AT 2471 lacking prephenate dehydrogenase activity. Our results revealed that the three recombinant enzymes are active. They all exhibit a high specificity toward arogenate and NADP, and have very similar kinetic properties. PMID- 11905964 TI - Deletion of the C-terminal 138 amino acids of the wheat FKBP73 abrogates calmodulin binding, dimerization and male fertility in transgenic rice. AB - Wheat FKBP73 (wFKBP73) belongs to the FK506-binding protein (FKBP) family which, in common with the cyclophilin and parvulin families, possesses peptidyl prolyl cis-trans isomerase (PPIase) activity. Wheat FKBP73 has been shown to contain three FKBP12-like domains, a tetratricopeptide repeat (TPR) via which it binds heat shock protein 90 and a calmodulin-binding domain (CaMbd). In this study we investigated: (1) the contribution of the N-terminal and C-terminal moieties of wFKBP73 to its biological activity by over-expression of the prolyl isomerase domains in transgenic rice, and (2) the biochemical characteristics of the C terminal moiety. The recombinant wFKBP73 was found to bind calmodulin via the CaMbd and to be present mainly as a dimer in solution. The dimerization was abrogated when 138 amino acids from the C-terminal half were deleted. Expression of the full-length FKBP73 produced fertile rice plants, whereas the expression of the peptidyl prolyl cis-trans isomerase domains in transgenic rice resulted in male-sterile plants. The male sterility was expressed at various stages of anther development with arrest of normal pollen development occurring after separation of the microspores from the tetrads. Although the direct cause of the dominant male sterility is not yet defined, we suggest that it is associated with a novel interaction of the prolyl isomerase domains with anther specific target proteins. PMID- 11905965 TI - Analysis of hypermethylation in the RPS element suggests a signal function for short inverted repeats in de novo methylation. AB - A repetitive DNA sequence (RPS) from Petunia hybrida had previously been shown to enhance expression variegation in petunia and tobacco and to carry a hot spot for de novo DNA methylation. Here we show that a strong de novo hypermethylation site is located within a palindromic segment of the RPS and present indirect evidence, based on sequence homologies to other repeat units within the RPS, for the formation of secondary structures at the methylation site in vivo. We demonstrate that the palindromic RPS element, which is moderately to highly repetitive in petunia, does not predominantly localise to constitutive heterochromatin. To test whether the RPS is subject to de novo methylation due to its repetitive nature or to intrinsic signals within the RPS, we integrated the RPS into the genome of Arabidopsis thaliana, a plant lacking homology to the RPS. Our data indicate that the palindromic element also acts as a de novo hypermethylation site in the non repetitive genomic background of Arabidopsis, strongly suggesting a signal function of the palindromic RPS element. PMID- 11905966 TI - Two flavonoid glucosyltransferases from Petunia hybrida: molecular cloning, biochemical properties and developmentally regulated expression. AB - Two flavonoid glucosyltransferases, UDP-glucose:flavonoid 3-0-glucosyltransferase (3-GT) and UDP-glucose: anthocyanin 5-O-glucosyltransferase (5-GT), are responsible for the glucosylation of anthocyani(di)ns to produce stable molecules in the anthocyanin biosynthetic pathway. The cDNAs encoding 3-GT and 5-GT were isolated from Petunia hybrida by hybridization screening with heterologous probes. The cDNA clones of 3-GT, PGT8, and 5-GT, PH1, encode putative polypeptides of 448 and 468 amino acids, respectively. A phylogenetic tree based on amino acid sequences of the family of glycosyltransferases from various plants shows that PGT8 belongs to the 3-GT subfamily and PH1 belongs to the 5-GT subfamily. The function of isolated cDNAs was identified by the catalytic activities for 3-GT and 5-GT exhibited by the recombinant proteins produced in yeast. The recombinant PGT8 protein could convert not only anthocyanidins but also flavonols into the corresponding 3-O-glucosides. In contrast, the recombinant PH1 protein exhibited a strict substrate specificity towards anthocyanidin 3-acylrutinoside, comparing with other 5-GTs from Perilla frutescens and Verbena hybrida, which showed broad substrate specificities towards several anthocyanidin 3-glucosides. The mRNA expression of both 3-GT and 5-GT increased in the early developmental stages of P. hybrida flower, reaching the maximum at the stage before flower opening. Southern blotting analysis of genomic DNA indicates that both 3-GT and 5-GT genes exist in two copies in P. hybrida, respectively. The results are discussed in relation to the molecular evolution of flavonoid glycosyltransferases. PMID- 11905968 TI - It takes a village: the caregiver's role in wound care. PMID- 11905969 TI - 5 questions--and answers--about hyperbaric oxygen therapy. PMID- 11905967 TI - Molecular characterization of nucleus-localized RNA-binding proteins from higher plants. AB - We report the isolation and characterization of genes from the higher plants Arabidopsis, spinach and tobacco which code for nucleus-localized RNA-binding proteins. Common features of these polypeptides are glycine/arginine-rich regions with several RGG repeats at their N- and C-termini, which are sufficient for RNA binding in northwestern assays. All polypeptides analysed contain two basic bipartite nuclear localization signals and translational fusions harbouring these regions with the beta-glucuronidase gene direct the fusion proteins into the nucleus. Nuclear localization was confirmed by cellular fractionation with a polyclonal antiserum raised against the over-expressed tobacco protein NtRGG1p. Two or three copies of related RGG genes appear to be present in the analysed organisms and the expression of some of them is regulated: a tobacco gene is light-regulated and a spinach gene is preferentially expressed in roots. Possible biological functions of this class of RNA-binding proteins as well as structure/function relationships related to the modular structure are discussed. PMID- 11905970 TI - The difference between albumin and prealbumin. PMID- 11905971 TI - A new Medicare Part B wound care policy. AB - Skin and wound care specialists should design an assessment tool that includes all the documentation required to prove medical necessity for the initial and continued use of NPWT, when appropriate for patients who are managing their wounds at home. In addition, specialists should supply the physician with the order form that is provided by the NPWT pump supplier. Finally, personnel should be trained to appropriately assess and document the need for NPWT. Once a Medicare Part B-covered patient at home qualifies for a NPWT pump and supplies, the supplier will deliver the equipment and supplies and will bill Medicare Part B directly. Payment will be 80% of the DMERC Fee Schedule. The patient will be charged 20% of the Medicare allowable fee. PMID- 11905972 TI - Are pressure ulcers preventable? A survey of experts. AB - OBJECTIVE: There is considerable debate regarding whether pressure ulcers can truly be prevented in nursing homes. New pressure ulcers are often taken as a sign of negligence that can lead to a lawsuit. This study sought to determine expert opinion regarding the preventability of pressure ulcers, the resources available to nursing homes for prevention, and the role of negligence lawsuits in pressure ulcer care. DESIGN: Survey mailed to a convenience sample of 98 experts in the field of pressure ulcer care. The survey contained 36 questions, most based on a 5-point Likert scale from "strongly agree"to"strongly disagree." Several questions asked respondents to rank items. RESULTS: Sixty-five of 92 surveys were completed (6 were returned but not completed) for a response rate of 71%. Sixty-two percent of respondents disagreed with the statement that all pressure ulcers are preventable. Only 5% said that nursing homes have adequate resources to prevent all pressure ulcers. Although most respondents disagreed that pressure ulcers are necessarily a sign of neglect and that nursing homes should be sued when a resident develops a pressure ulcer, 38% agreed with the concept that lawsuits are an appropriate way to stimulate improvement in nursing home care. CONCLUSION: The results of this survey demonstrated divergent expert opinion on whether pressure ulcers are preventable. The role of regulations and litigation in pressure ulcer prevention needs to be further defined. PMID- 11905974 TI - Wound care products: how to choose. PMID- 11905973 TI - Translating pressure ulcer guidelines into practice: it's harder than it sounds. AB - OBJECTIVE: To determine the cost-effectiveness of a guideline-based pressure ulcer prevention protocol over time. DESIGN: Retrospective and prospective quasi experimental longitudinal design. Costs are presented from the long-term-care facility perspective. Data collection occurred for 3 periods: the first 6 months of 1994 (prior to protocol implementation), the first 6 months of 1995 (immediately following implementation) and the first 6 months of 1997 (2 years following implementation). SETTING: 77-bed long-term-care facility PATIENTS/PARTICIPANTS: Subjects were ulcer-free facility residents at the start of each data collection period. There were 69 subjects in the 1994 sample, 63 in the 1995 sample, and 71 in the 1997 sample. INTERVENTIONS: A guideline-based pressure ulcer prevention protocol was implemented during the last 3 months of 1994. The protocol consisted of specific policies for pressure ulcer prevention and treatment, intensive staff education on pressure ulcer care, and monitoring with regular performance feedback to staff. MEASUREMENTS AND MAIN RESULTS: Time to ulcer development varied among the 3 groups (log rank = 8.81, P = .01), with longer time to ulcer development in 1995 compared with 1994; no difference was seen between 1997 and 1994. The time for ulcers to heal decreased over the 3 years (log rank = 9.49, P <.01), with ulcer healing time being shorter in 1995 and 1997 compared with 1994. Total costs were unchanged during the 3 years (F = 0.2, P =.81). Costs of treatment declined significantly from 1994 to 1995 and 1997 (F = 5.5, P <.01) and costs of prevention increased significantly from 1994 to 1995 and 1997(F = 15, P <.01). From 1994 to 1997, the cost for 1 day of ulcer free life was $3.50. CONCLUSIONS: Implementation of a pressure ulcer prevention protocol showed mixed results. Initial reductions in pressure ulcer incidence were lost over time. Clinical results of ulcer treatment, however, improved and treatment costs fell during the 3 years. Implementation of preventive programs poses a major leadership challenge. PMID- 11905975 TI - An interview with Ted Kennedy, Jr. PMID- 11905976 TI - The patient.com. PMID- 11905977 TI - A randomized control trial to evaluate pressure-reducing seat cushions for elderly wheelchair users. AB - OBJECTIVE: To determine if the use of pressure-reducing wheelchair cushions for elderly nursing home resident wheelchair users who are at high risk for developing sitting-acquired pressure ulcers would result in a lower incidence rate of pressure ulcers, a greater number of days until ulceration, and lower peak interface pressures compared with the use of convoluted foam cushions over a 12-month period. To determine the feasibility of conducting a subsequent full scale definitive trial to evaluate the use of pressure-reducing seat cushions for elderly nursing home resident wheelchair users. DESIGN: Randomized control trial SETTING: 2200-bed skilled nursing facilities (1 suburban and 1 urban academic medical center) PATIENTS: 32 male and female at-risk nursing home residents who were wheelchair users > or = 65 years of age. Participants had Braden Scale scores < or = 18, Braden Activity and Mobilitysubscale scores < or = 5, no sitting surface pressure ulcers, and a daily wheelchair sitting tolerance of more than 6 hours. All met criteria for using the ETAC Twin wheelchair. INTERVENTIONS: Seating evaluation with pressure-mapping and subsequent seating prescription. Subjects were assigned to either a foam (n=17) or pressure-reducing cushion (n=15) group and weekly assessments of skin and pressure ulcer risk were made. MAIN OUTCOME MEASURES: Incidence of pressure ulcers, days to ulceration, and peak interface pressure. MAIN RESULTS: At a 95% confidence interval, a 2-tailed analysis showed no differences between the FOAM and pressure-reducing cushion groups for pressure ulcer incidence, total days to pressure ulcer, or initial peak interface pressure. Pressure-reducing cushions were more effective in preventing sitting-acquired (ischial) pressure ulcers (P<.005). Higher interface pressures were associated with a higher incidence of pressure ulcers (P<.001). CONCLUSIONS: A definitive randomized control multicenter cushion trial is feasible with a sample size of 50 to 100 per study group. In the definitive trial, the definition of sitting-acquired pressure ulcers should be limited to lesions occurring over the ischial tuberosities. PMID- 11905978 TI - Clinical skin temperature measurement to predict incipient pressure ulcers. AB - OBJECTIVE: To evaluate temperature differences between areas of erythema and surrounding healthy tissue to determine whether clinical temperature measurement of sites at risk for pressure ulcer development could be used to indicate tissue damage. To validate the temperature portion of the National Pressure Ulcer Advisory Panel's new Stage I pressure ulcer definition. DESIGN: Repeated measures design. SETTING: Acute rehabilitation hospital. SUBJECTS: 65 outpatients and inpatients presenting with pressure-induced erythema at areas at risk for pressure ulcer development. The subjects were primarily non-ambulatory and exhibited a range of skin pigmentation and disabilities, including spinal injury, multiple sclerosis, and lower-limb amputations. MAIN RESULTS: The temperature and appearance of 80 pairs of erythematic and control sites were documented. Sites were considered to have equal temperatures if the difference was within plus or minus 1.0 degree F. Fifteen percent (n = 12) of the erythematic sites were the same temperature as the surrounding tissue, 23% (n = 18) of the erythematic sites were cooler than the control sites, and 63% (n = 50) were warmer. CONCLUSION: Both increased and decreased temperature differences can be used to indicate reactive hyperemia or a Stage I pressure ulcer, but a tissue integrity problem may still exist despite the absence of a temperature difference. PMID- 11905979 TI - 9 clinical cases of nonhealing pressure ulcers in patients with spinal cord injury treated with an anabolic agent: a therapeutic trial. AB - Anabolic steroid agents may potentially promote wound healing in individuals with spinal cord injury who have long-standing wounds. Nine patients hospitalized on the Spinal Cord Injury Service of the VA Medical Center, Bronx, NY, received treatment with an anabolic agent and amino acid supplement. Selection of patients was based on having at least 1 of the following criteria: (1) nonhealing pressure ulcer of at least 2 months' duration with no change or worsening status, and/or (2) full-thickness pressure ulcer through fascia into muscle, tendon, or bone. Previous and current pressure ulcer histories were determined by review of the hospital records when available and/or self-reporting by the patient. Eight of 9 patients had nonhealing wounds of 2 months' to 5 years' duration. One patient was included because of having a large, full-thickness pressure ulcer of reportedly 2 weeks' duration. Except for 1 Stage III pressure ulcer, all others had Stage IV pressure ulcers.Three patients had documented weight loss (>10% of total body weight), 3 had no recent weight loss, and 3 refused to be weighed. Patients were treated from 1 to 12 months with oxandrolone (20 mg/day) and glutamine (20 g/day). Eight of 9 patients completely healed: 3 after 3 months, 2 after 4 months, 1 after 6 months, and 2 after 12 months of treatment. One patient was discharged against medical advice after 1 month of treatment with a healing wound and was lost to follow-up. Although clinical case studies have limited usefulness for determining effectiveness of drug therapy on wound healing, these initial observations of successful treatment with an anabolic agent and amino acid supplement have been encouraging. PMID- 11905980 TI - Arterial vs venous ulcers: diagnosis and treatment. PMID- 11905981 TI - Management of a venous ulcer: a case study approach. PMID- 11905982 TI - Diagnosing pyoderma gangrenosum. PMID- 11905983 TI - Anabolic agents and wound healing. PMID- 11905984 TI - 5 questions--and answers--about electrical stimulation. PMID- 11905985 TI - Relevance of different striatal markers in assessment of the MPP+-induced dopaminergic nigrostriatal injury in rat. AB - Many striatal dopaminergic markers are available for estimating the degree of the nigrostriatal lesion by MPTP/MPP+, but the changes of these markers are not perfectly matched. In this study we investigated different striatal markers and determined which ones closely reflected the nigrostriatal alteration. The in vivo binding of (E)-N-(3-iodoprop-2-enyl)-2-beta-carbomethoxy-3beta-(4' methylphenyl)nortropane (PE2I), a selective and potent inhibitor of the neuronal dopamine transporter (DAT) was considered as the reference index of injury of striatal dopaminergic nerve-endings. Rats received a 10-microg MPP+ injection in the right substantia nigra and were killed at 7 days after lesion. The results were as follows: (i) a decrease (66%) of the biodistribution of [125I]PE2I; (ii) a great reduction of the DAT expression measured by the binding of [125I]PE2I in striatal membranes (Bmax decreased by 54%) and in cerebral slices (88%); (iii) an 80% inhibition of the vesicular monoamine transporter expression revealed by the binding of [3H]dihydrotetrabenazine in cerebral slices; (iv) a robust decrease in the quantity of DA and its metabolites (about 50-60%); (v) a slight modification of the DAT activity with a decreased number of functional sites (Vmax decreased by 12%, p < 0.05) without change of the affinity in striatal synaptosomes. Among these markers the binding of [125I]PE2I in membrane homogenates and the content of DA, and its metabolites, in striatum could be the most relevant in vitro indexes of the degenerative state of the nigrostriatal pathway after MPP+ lesion. PMID- 11905986 TI - Neuronal storage of histamine in the brain and tele-methylimidazoleacetic acid excretion in portocaval shunted rats. AB - Rats with portocaval anastomosis (PCA), an animal model of hepatic encephalopathy (HE), have very high brain histamine concentrations. Our previous studies based on a biochemical approach indicated histamine accumulation in the neuronal compartment. In this study, immunohistochemical evidence is presented which further supports the amine localization in histaminergic neurons. These neurons become pathological in appearance with cisternae frequently seen along histaminergic fibres in many brain areas, including the hypothalamus, amygdala, substantia nigra and cerebral cortex. Such formations were not observed in sham operated animals. The neuronal deposition is predominant, and unique for histamine. It serves as a mechanism to counterbalance excessive brain neurotransmitter formation evoked by PCA. However, there are other mechanisms. The data provided here show that there is also a significant increase in histamine catabolism in the shunted rats, as reflected by both the higher brain N tele-methylhistamine (t-MeHA) concentration and urinary excretion of N-tele methylimidazoleacetic acid (t-MelmAA), a major brain histamine end product. The stomach, in addition to the brain, is a site of enhanced histamine synthesis in portocavally shunted subjects. After gastrectomy or food deprivation to eliminate the contribution of the stomach, shunted rats excrete significantly more t MelmAA, implying the role of the CNS. This last finding suggests that under strictly defined conditions, namely in parenterally fed HE patients with abnormal plasma L-histidine, the measurement of urinary t-MelmAA might provide valuable information concerning putative brain histaminergic activity. PMID- 11905987 TI - Modulation of the GABA(A)-gated chloride channel by reactive oxygen species. AB - The accumulation of reactive oxygen species during cellular injury leads to oxidative stress. This can have profound effects on ionic homeostasis and neuronal transmission. Gamma-aminobutyric acid (GABA) neurotransmission is sensitive to reactive oxygen species, but most studies have indicated that this is due to alterations in GABA release. Here, we determined whether reactive oxygen species can alter GABA(A) receptor-gated Cl- channels in the adult hippocampus. First, we measured the effects of hydrogen peroxide on intracellular Cl- using UV laser scanning confocal microscopy and the Cl(-)-sensitive probe, 6 methoxy-N-ethylquinolium iodide (MEQ). Superfusion of adult rat hippocampal slices with hydrogen peroxide for 10 min decreased MEQ fluorescence (elevation in [Cl-]i) significantly in area CA1 pyramidal cell soma. Alterations in [Cl-]i were prevented by the vitamin E analog Trolox, an antioxidant that scavenges free radicals. After exposure of slices to hydrogen peroxide, the ability of the GABA agonist muscimol to increase [Cl-]i was attenuated. To determine if GABA(A) receptors were sensitive to oxidative insults, the effect of hydrogen peroxide on the binding of [35S]t-butylbicyclophosphorothionate (TBPS) to GABA-gated Cl- channels was measured using receptor autoradiography and homogenate binding assays. Hydrogen peroxide inhibited [35S]TBPS binding in a regionally selective manner, with the greatest inhibition in cerebral cortex, hippocampus and striatum, areas vulnerable to oxidative stress. Similarly, xanthine and xanthine oxidase, which generate superoxide radicals, reduced [35S]TBPS binding in these regions. The effect of hydrogen peroxide on [35S]TBPS binding was non-competitive and was prevented by Trolox and the iron chelator, deferoxamine. We conclude that reactive oxygen species may compromise GABA(A)-mediated neuronal inhibition via interaction with pre and postsynaptic sites. A reduction in GABA(A)-gated Cl- channel function during periods of oxidative stress may contribute to the development of neuronal damage. PMID- 11905988 TI - The distribution of the anti-HIV drug, 2'3'-dideoxycytidine (ddC), across the blood-brain and blood-cerebrospinal fluid barriers and the influence of organic anion transport inhibitors. AB - The brain and CSF distribution of the HIV reverse transcriptase inhibitor, 2'3' dideoxycytidine (ddC), was investigated by the in situ brain perfusion and isolated incubated choroid plexus methods in the guinea pig. Multiple-time brain perfusions indicated that the distribution of [3H]ddC to the brain and CSF was low and the unidirectional rate constant (K(in)) for the brain uptake of this nucleoside analogue (0.52 +/- 0.10 microL/min/g) was not significantly different to that for the vascular marker, [14C]mannitol (0.44 +/- 0.09 microL/min/g). The influence of unlabelled ddC, six organic anion transport inhibitors and 3'-azido 3'-deoxythymidine (AZT) on the CNS uptake of [3H]ddC was examined in situ and in vitro. ddC, probenecid and 2,4-dichlorophenoxyacetic acid altered the distribution of [3H]ddC into the brain and choroid plexuses, indicating that the limited distribution of [3H]ddC was a result of an organic anion efflux transporter, in addition to the low lipophilicity of this drug (octanol-saline partition coefficient, 0.047 +/- 0.001). The CNS distribution was also sensitive to p-aminohippurate and deltorphin II, but not digoxin, suggesting the involvement of organic anion transporters (OAT1/OAT3-like) and organic anion transporting polypeptides (OATP1/OATPA-like). AZT did not effect the accumulation of [3H]ddC, indicating that when these nucleoside analogues are used in anti-HIV combination therapy, the CNS distribution of ddC is unchanged. PMID- 11905989 TI - Mitochondria regulate Ca2+ wave initiation and inositol trisphosphate signal transduction in oligodendrocyte progenitors. AB - Mitochondria in oligodendrocyte progenitor cells (OPs) take up and release cytosolic Ca2+ during agonist-evoked Ca2+ waves, but it is not clear whether or how they regulate Ca2+ signaling in OPs. We asked whether mitochondria play an active role during agonist-evoked Ca2+ release from intracellular stores. Ca2+ puffs, wave initiation, and wave propagation were measured in fluo-4 loaded OP processes using linescan confocal microscopy. Mitochondrial depolarization, measured by tetramethyl rhodamine ethyl ester (TMRE) fluorescence, accompanied Ca2+ puffs and waves. In addition, waves initiated only where mitochondria were localized. To determine whether energized mitochondria were necessary for wave generation, we blocked mitochondrial function with the electron transport chain inhibitor antimycin A (AA) in combination with oligomycin. AA decreased wave speed and puff probability. These effects were not due to global changes in ATP. We found that AA increased cytosolic Ca2+, markedly reduced agonist-evoked inositol trisphosphate (IP3) production, and also enhanced phosphatidylinositol 4,5-bisphosphate (PIP2) binding to the Ca2+ dependent protein gelsolin. Thus, the reduction in puff probability and wave speed after AA treatment may be explained by competition for PIP2 between phospholipase C and gelsolin. Energized mitochondria and low cytosolic Ca2+ concentration may be required to maintain PIP2, a substrate for IP3 signal transduction. PMID- 11905990 TI - Neurotoxic mechanisms triggered by Alzheimer's disease-linked mutant M146L presenilin 1: involvement of NO synthase via a novel pertussis toxin target. AB - While it has been reported that familial Alzheimer's disease (FAD)-linked mutants of amyloid precursor protein (APP) and presenilin (PS)2 induce neuronal cytotoxicity in a manner sensitive to antioxidant and pertussis toxin (PTX), little of the mechanism for PS1-mediated neuronal cell death has been characterized. We previously found that multiple mechanisms, different in detail, underlie cytotoxicities by two FAD-linked mutants of APP, using neuronal cells with an ecdysone-controlled expression system. Here we report that this system revealed that (i) low expression of FAD-linked M146L-PS1 caused neuronal cell death, whereas that of wild-type (wt)PS1 did not; (ii) mutation-specific cytotoxicity by M146L-PS1 was sensitive to antioxidant glutathione-ethyl-ester and resistant to Ac-DEVD-CHO; (iii) cytotoxicity by higher expression of wtPS1 was resistant to both; and (iv) cytotoxicity by M146L-PS1 was inhibited by PTX. It was also highly likely that the involved superoxide-generating enzyme was nitric oxide synthase (NOS), and that the PTX-sensitive cytotoxic signal by M146L PS1 was mediated by none of the G(i/o) proteins. We conclude that M146L-PS1 activates a NOS-mediated cytotoxic pathway via a novel PTX target. PMID- 11905991 TI - Neuroprotective properties of cannabinoids against oxidative stress: role of the cannabinoid receptor CB1. AB - Neuroprotective effects have been described for many cannabinoids in several neurotoxicity models. However, the exact mechanisms have not been clearly understood yet. In the present study, antioxidant neuroprotective effects of cannabinoids and the involvement of the cannabinoid receptor 1 (CB1) were analysed in detail employing cell-free biochemical assays and cultured cells. As it was reported for oestrogens that the phenolic group is a lead structure for antioxidant neuroprotective effects, eight compounds were classified into three groups. Group A: phenolic compounds that do not bind to CB1. Group B: non phenolic compounds that bind to CB1. Group C: phenolic compounds that bind to CB1. In the biochemical assays employed, a requirement of the phenolic lead structure for antioxidant activity was shown. The effects paralleled the protective potential of group A and C compounds against oxidative neuronal cell death using the mouse hippocampal HT22 cell line and rat primary cerebellar cell cultures. To elucidate the role of CB1 in neuroprotection, we established stably transfected HT22 cells containing CB1 and compared the protective potential of cannabinoids with that observed in the control transfected HT22 cell line. Furthermore, oxidative stress experiments were performed in cultured cerebellar granule cells, which were derived either from CB1 knock-out mice or from control wild-type littermates. The results strongly suggest that CB1 is not involved in the cellular antioxidant neuroprotective effects of cannabinoids. PMID- 11905992 TI - The alpha7 nicotinic receptors in human fetal brain and spinal cord. AB - The alpha7 nicotinic acetylcholine receptor subtype is believed to be involved in the regulation of neuronal growth, differentiation and synapse formation during the development of the human brain. In this study the expression of the alpha7 nicotinic acetylcholine receptor was investigated in human fetal brain and spinal cord of 5-11 weeks gestational age. Both the specific binding of [125I]alpha bungarotoxin to prenatal brain membranes and the expression of alpha7 mRNA were significantly higher in the pons, medulla oblongata, mesencephalon and spinal cord of 9-11 weeks gestational age compared with cerebellum, cortex and subcortical forebrain. A significant positive correlation between gestational age and the expression of alpha7 mRNA was observed in all brain regions except cortex. A positive correlation was also observed between the gestational age and the [125I]alpha-bungarotoxin binding in the pons, medulla oblongata, mesencephalon, and cerebellum. Consequently, a significant relationship between the alpha7 mRNA levels and the binding sites for [125I]alpha-bungarotoxin was found in the fetal brain. The increasing levels of the alpha7 nicotinic acetylcholine receptor during the first trimester support the important role of nAChRs for the development of the central nervous system. PMID- 11905993 TI - Metabolic changes in the basal ganglia of patients with Huntington's disease: an in situ hybridization study of cytochrome oxidase subunit I mRNA. AB - On the basis of the functional model of the basal ganglia developed in the 1980s and the neuropathological findings in Huntington's disease (HD), changes in the neuronal activity of the basal ganglia have previously been proposed to explain the abnormal movements observed in this pathology. In particular, it has been stated that the neurodegenerative process affecting the basal ganglia in the disease should provoke a hypoactivity in the internal segment of the pallidum (GPi) that could explain choreic movements observed in the disease. To test this functional hypothesis, we performed an in situ hybridization study on control and HD brains postmortem, taking cytochrome oxidase subunit I (COI) mRNAs expression as index of neuronal activity. As most of the HD patients studied were under chronic neuroleptic (NL) treatment, we also studied the brains of non-HD patients under chronic NL treatment. Our results show that in HD brain the number of neurons expressing COI mRNA tends to be lower in the striatum, GPe and GPi, suggesting a severe involvement of these structures during the neurodegenerative process. Moreover, COI mRNA level of expression was markedly reduced within neurons of the putamen and GPe. Surprisingly, COI mRNA expression was not modified in the GPi in HD brains compared with controls. This paradoxical result in the GPi may be explained by the antagonistic effect of GPe hypoactivity and the degenerative process involving neurons of GPi. Our results indicate that the functional modifications, and consequently the pathophysiology of abnormal movements, observed in HD basal ganglia are more complex than expected from the currently accepted model of the basal ganglia organization. PMID- 11905994 TI - Substrate specificity, inhibition and enzymological analysis of recombinant human glutamate carboxypeptidase II. AB - Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a membrane peptidase expressed in a number of tissues such as kidney, prostate and brain. The brain form of GCPII (also known as NAALADase) cleaves N-acetyl-aspartyl glutamate to yield free glutamate. Animal model experiments show that inhibition of GCPII prevents neuronal cell death during experimental ischaemia. GCPII thus represents an important target for the treatment of neuronal damage caused by excess glutamate. In this paper we report expression of an extracellular portion of human glutamate carboxypeptidase II (amino acids 44-750) in Drosophila Schneider's cells and its purification to homogeneity. A novel assay for hydrolytic activity of recombinant human GCPII (rhGCPII), based on fluorimetric detection of released alpha-amino groups was established, and used for its enzymological characterization. rhGCPII does not show dipeptidylpeptidase IV-like activity assigned to the native form of the enzyme previously. Using a complete set of protected dipeptides, substrate specificity of rhGCPII was elucidated. In addition to the previously described substrates, four novel compounds, Ac-Glu Met, Ac-Asp-Met and, surprisingly, Ac-Ala-Glu and Ac-Ala-Met were identified as substrates for GCPII, and their respective kinetic constants determined. The glycosylation of rhGCPII was found indispensable for the enzymatic activity. PMID- 11905995 TI - Increased mitochondrial antioxidative activity or decreased oxygen free radical propagation prevent mutant SOD1-mediated motor neuron cell death and increase amyotrophic lateral sclerosis-like transgenic mouse survival. AB - The molecular mechanisms of selective motor neuron degeneration in human amyotrophic lateral sclerosis (ALS) disease remain largely unknown and effective therapies are not currently available. Mitochondrial dysfunction is an early event of motor neuron degeneration in transgenic mice overexpressing mutant superoxide dismutase (SOD)1 gene and mitochondrial abnormality is observed in human ALS patients. In an in vitro cell culture system, we demonstrated that infection of mouse NSC-34 motor neuron-like cells with adenovirus containing mutant G93A-SOD1 gene increased cellular oxidative stress, mitochondrial dysfunction, cytochrome c release and motor neuron cell death. Cells pretreated with highly oxidizable polyunsaturated fatty acid elevated lipid peroxidation and synergistically exacerbated motor neuron-like cell death with mutant G93A-SOD1 but not with wild-type SOD1. Similarly, overexpression of mitochondrial antioxidative genes, MnSOD and GPX4 by stable transfection significantly increased NSC-34 motor neuron-like cell resistance to mutant SOD1. Pre-incubation of cells with spin trapping molecule, 5',5'-dimethylpryrroline-N-oxide (DMPO), prevented mutant SOD1-mediated mitochondrial dysfunction and cell death. Furthermore, treatment of mutant G93A-SOD1 transgenic mice with DMPO significantly delayed paralysis and increased survival. These findings suggest a causal relationship between enhanced oxidative stress and mutant SOD1-mediated motor neuron degeneration, considering that enhanced oxygen free radical production results from the SOD1 structural alterations. Molecular approaches aimed at increasing mitochondrial antioxidative activity or effectively blocking oxidative stress propagation can be potentially useful in the clinical management of human ALS disease. PMID- 11905996 TI - PC12nnr5 cells expressing TrkA receptors undergo morphological but not cholinergic phenotypic differentiation in response to nerve growth factor. AB - We investigated mechanisms underlying nerve growth factor-mediated morphological differentiation and expression of cholinergic neuronal phenotype. In PC12, but not PC12nnr5 cells, nerve growth factor induces neurite-like outgrowths and enhances cholinergic phenotype; stable expression of TrkA receptors in nnr5 cells (called B5P cells) restores morphological differentiation but not expression of choline acetyltransferase. Transfection with an AP-1 luciferase reporter gene revealed that PC12 but not B5P cells expressed nerve growth factor-induced functional AP-1 activity. RT-PCR analysis of nerve growth factor-mediated changes in AP-1 transcription factors showed rapid increases in c-fos and junB mRNA in PC12 and B5P cells, while increases in c-jun were small. Using DNA-protein gel shift assays we determined that nerve growth factor stimulates AP-1 binding in both PC12 and B5P cells, and identified c-Fos, FosB, Fra-1, Fra-2, c-Jun, JunB and JunD in AP-1 complexes. In Fos/Jun functional luciferase reporter assays, nerve growth factor stimulated phosphorylation of c-Fos in both PC12 and B5P cells, but phosphorylation of c-Jun only in PC12, and not in B5P cells. These data indicate that mechanisms relating to AP-1 transcription factor complexes underlying nerve growth factor-mediated enhancement of cholinergic gene expression may differ from those required for morphological differentiation. PMID- 11905997 TI - Nicotine activates the extracellular signal-regulated kinase 1/2 via the alpha7 nicotinic acetylcholine receptor and protein kinase A, in SH-SY5Y cells and hippocampal neurones. AB - Neuronal nicotinic acetylcholine receptors (nAChR) can modulate many cellular mechanisms, such as cell survival and memory processing, which are also influenced by the serine/threonine protein kinases ERK1/2. In SH-SY5Y cells and hippocampal neurones, nicotine (100 microM) increased the activity of ERK1/2. This effect was Ca2+ dependent, and prevented by the alpha7 nAChR antagonist alpha-bungarotoxin (alpha-Bgt) and an inhibitor (PD98059) of the upstream kinase MEK. To determine the intervening steps linking Ca2+ entry to MEK-ERK1/2 activation, inhibitors of Ca2+-dependent kinases were deployed. In SH-SY5Y cells, selective blockers for PKC (Ro 31-8220), CaM kinase II (KN-62) or PI3 kinase (LY 294002) failed to inhibit the nicotine-evoked increase in ERK1/2 activity. In contrast, two structurally different inhibitors of PKA (KT 5720 and H-89) completely prevented the nicotine-dependent increase in ERK1/2 activity. Inhibition of the nicotine-evoked increase in ERK1/2 activity by H-89 was also observed in hippocampal cultures. Down stream of PKA, the activity of B-Raf was significantly decreased by nicotine in SH-SY5Y cells, as determined by direct measurement of MEK1 phosphorylation or in vitro kinase assays, whereas the modulation of MEK1 phosphorylation by Raf-1 tended to increase. Thus, this study provides evidence for a novel signalling route coupling the stimulation of alpha7 nAChR to the activation of ERK1/2, in a Ca2+ and PKA dependent manner. PMID- 11905998 TI - Mitochondrial permeability transition and calcium dynamics in striatal neurons upon intense NMDA receptor activation. AB - Deregulation of the intracellular Ca2+ homeostasis by NMDA receptor activation leads to neuronal cell death. Induction of the mitochondrial permeability transition pore (MPT) by Ca2+ is a critical event in mediating cell death. In this study, we used fluorescent Ca2+ indicators to investigate the effect of high concentrations of NMDA on cytosolic and mitochondrial Ca2+ concentrations ([Ca2+]c and [Ca2+]m, respectively) in cultured striatal neurons. Exposure to NMDA resulted in an immediate, sustained increase in [Ca2+]c followed by a secondary increase in [Ca2+]c. This second increase of [Ca2+]c was prevented by pretreatment with N-methyl-valine-4-cyclosporin (NMV-Cys). Exposure of neurons to NMDA also resulted in an increase in [Ca2+]m that was followed by a precipitous decrease in the rhod-2 signal. This decrease followed the time frame of the secondary increase in [Ca2+]c. Preincubation of the neurons with NMV-Cys prevented the decrease in rhod-2 fluorescence. These dynamic changes in the rhod 2 signal and [Ca2+]m in response to NMDA were confirmed by using confocal microscopy. The presented results indicate that MPT can be detected in living neurons using fluorescent Ca2+ indicators, which would allow the study of the physiological role of MPT in cell death. PMID- 11905999 TI - Dietary restriction enhances neurotrophin expression and neurogenesis in the hippocampus of adult mice. AB - The adult brain contains small populations of neural precursor cells (NPC) that can give rise to new neurons and glia, and may play important roles in learning and memory, and recovery from injury. Growth factors can influence the proliferation, differentiation and survival of NPC, and may mediate responses of NPC to injury and environmental stimuli such as enriched environments and physical activity. We now report that neurotrophin expression and neurogenesis can be modified by a change in diet. When adult mice are maintained on a dietary restriction (DR) feeding regimen, numbers of newly generated cells in the dentate gyrus of the hippocampus are increased, apparently as the result of increased cell survival. The new cells exhibit phenotypes of neurons and astrocytes. Levels of expression of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT 3) are increased by DR, while levels of expression of high-affinity receptors for these neurotrophins (trkB and trkC) are unchanged. In addition, DR increases the ratio of full-length trkB to truncated trkB in the hippocampus. The ability of a change in diet to stimulate neurotrophin expression and enhance neurogenesis has important implications for dietary modification of neuroplasticity and responses of the brain to injury and disease. PMID- 11906000 TI - Functional effects of tau gene mutations deltaN296 and N296H. AB - Mutations in the tau gene cause frontotemporal dementia and parkinsonism linked to chromosome-17 (FTDP-17). Functionally, about half of the known mutations increase the alternative mRNA splicing of exon 10 of the tau gene, resulting in the overproduction of tau isoforms with four microtubule-binding repeats. The other mutations reduce the ability of tau to interact with microtubules, with some mutations also increasing the propensity of tau to assemble into filaments. Here we have examined the functional effects of the recently described tau gene mutations deltaN296 and N296H. Both mutations reduced the ability of tau to promote microtubule assembly, without having a significant effect on tau filament formation. By exon trapping, they increased the splicing of exon 10. DeltaN296 and N296H thus define a class of tau mutations with effects at both the RNA and the protein level. PMID- 11906001 TI - Glucose deprivation and hypoxia increase the expression of the GLUT1 glucose transporter via a specific mRNA cis-acting regulatory element. AB - The expression of the glucose transporter type-1 (GLUT1) gene is up-regulated in hypoxia and glucose deprivation. A 10 nucleotide (nt) cis-acting regulatory element (CAE), which is located within nt 2181-2190 of the GLUT1 3'-untranslated region (CAE2181-2190), increases the expression of a GLUT1-luciferase reporter gene and decreases its mRNA decay. The present study investigated the role of the GLUT1 CAE2181-2190 in glucose deprivation and hypoxia using stable transfectants. Glucose and O2 deprivation produced a marked increase in the expression of the GLUT1 reporter gene carrying the CAE2181-2190, and this effect was additive. Glucose deprivation and/or hypoxia induced no significant changes in the expression of the reporter gene wherein the GLUT1 CAE2181-2190 was site-directed deleted. Data presented here suggest that the GLUT1 CAE2181-2190 participates in the increase of GLUT1 gene expression in glucose deprivation and hypoxia. PMID- 11906002 TI - Bioinformatics of large-scale protein interaction networks. AB - We survey recent techniques for construction and prediction of large-scale protein interaction networks, focusing on computational processing steps. Special emphasis is placed on critical assessment of data completeness and reliability of the various approaches. Once built, protein interaction networks can be used for functional annotation or to generate higher-level biological hypotheses on pathways. PMID- 11906003 TI - From the decline and fall of protein chemistry to proteomics. PMID- 11906005 TI - ProteinChip clinical proteomics: computational challenges and solutions. AB - ProteinChip technology, a suite of analytical tools that includes retentate chromatography, on-chip protein characterization, and multivariate analysis, allows researchers to examine patterns ofprotein expression and modification. Based on the surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) approach, ProteinChip technology has been pioneered by researchers at Ciphergen Biosystems (Fremont, CA, USA), as well as by users of Ciphergen's commercial embodiment of this technology the ProteinChip Biomarker System. This report will begin with a background of the technology and describe its applications in clinical proteomics and will then conclude with a discussion of tools and strategies to mine the large amounts of data generated during the course of a typical clinical proteomics study. PMID- 11906004 TI - The use of chemical design tools to transform proteomics data into drug candidates. AB - The Human Genome Project has fueled the massive information-driven growth of genomics and proteomics and promises to deliver new insights into biology and medicine. Since proteins represent the majority of drug targets, these molecules are the focus of activity in pharmaceutical and biotechnology organizations. In this article, we describe the processes by which computational drug design may be used to exploit protein structural information to create virtual small molecules that may become novel medicines. Experimental protein structure determination, site exploration, and virtual screening provide a foundation for small molecule generation in silico, thus creating the bridge between proteomics and drug discovery. PMID- 11906006 TI - Informatic tools for proteome profiling. AB - In recent years, the practice of proteomics research has experienced a dramatic shift within the pharmaceutical and biotechnology industry with the widespread implementation of novel applications. The areas of interest extend all the way from discovery of novel drug, vaccine, and diagnostic targets, characterization of protein-based products, toxicology, and identification of surrogate markers of activity in clinical research, to the ability to provide information on the mechanisms of drug action. The power of two-dimensional gel electrophoresis as well as advances in mass spectrometric techniques combined with sequence database correlation have enabled speed and accuracy in identification of proteins in complex mixtures. This article surveys currently available software and informatic tools related to these methods for proteome profiling. The broad acceptance of these technologies, however, has not been accompanied by significant advances in the informatics and software tools necessary to support the analysis and management of the massive amounts of data generated in the process. In this context, this article also discusses the importance of relational databases for protein identification data management. PMID- 11906007 TI - Structural proteomics: methods in deriving protein structural information and issues in data management. AB - Structural proteomics is an emerging paradigm that is gaining importance in the post-genomic era as a valuable discipline to process the protein target information being deciphered. The field plays a crucial role in assigning function to sequenced proteins, defining pathways in which the targets are involved, and understanding structure-function relationships of the protein targets. A key component of this research sector is accessing the three dimensional structures of protein targets by both experimental and theoretical methods. This then leads to the question of how to store, retrieve, and manipulate vast amounts of sequence (1-D) and structural (3-D) information in a relational format so that extensive data analysis can be achieved. We at SBI have addressed both of these fundamental requirements of structural proteomics. We have developed an extensive collection of three-dimensional protein structures from sequence data and have implemented a relational architecture for data management. In this article we will discuss our approaches to structural proteomics and the tools that life science researchers can use in their discovery efforts. PMID- 11906008 TI - Raw data to knowledge warehouse in proteomic-based drug discovery: a scientific data management issue. PMID- 11906009 TI - Market opportunity in computational proteomics. AB - The current exuberance on the potential of proteomics as a means to deploy the wealth of the human genome is expected to last into the coming years. Unlike the genome, a finite entity with a fixed number of base pairs of the genetic material, the proteome is "plastic", changing throughout growth and development and environmental stresses, as well as in pathological situations. Our proteomes change over time, and therefore there is no one proteome; the proteome is for practical purposes an infinite entity. It is therefore crucial to build systems that are capable of manipulating the information content that is the proteome, thence the need for computational proteomics as a discipline. In this Market View article, we present the industry landscape that is emerging in the computational proteomics space. This space is still in its infancy and for the most part undefined; therefore we seek to present the market opportunity in informatics in the drug discovery space and then extend that to an examination of industry trends in proteomics. Thus, the gestalt is a set of predictions as to the evolution of the landscape in computational proteomics over the coming years. PMID- 11906010 TI - Summary of Pneumocystis research presented at the 7th International Workshop on Opportunistic Protists. PMID- 11906011 TI - DNA analysis of Pneumocystis infecting a Cavalier King Charles spaniel. PMID- 11906012 TI - Pneumocystis carinii infection in red-bellied tamarins and cynomolgus monkeys, and the characterization of the mitochondrial large subunit ribosomal RNA gene of Pneumocystis carinii. PMID- 11906013 TI - Three new karyotype forms of Pneumocystis carinii f. sp. carinii identified by contoured clamped homogeneous electrical field (CHEF) electrophoresis. PMID- 11906014 TI - Survey of sera from encephalitis patients for Balamuthia mandrillaris antibody. PMID- 11906015 TI - Spatial analysis on the occurrence of Pneumocystis carinii in the shrew Notiosorex crawfordi in fragmented landscape in southern California. PMID- 11906016 TI - Parallel phylogenies of Pneumocystis species and their mammalian hosts. AB - The single name Pneumocystis carinii consists of an heterogeneous group of specific fungal organisms that colonize a very wide range of mammalian hosts. In the present study, mitochondrial large subunit (mtLSU) and small subunit (mtSSU) rRNA sequences of P. carinii organisms from 24 different mammalian species were compared. The mammals were included in six major groups: Primates (12 species), Rodents (5 species), Carnivores (3 species), Bats (1 species), Lagomorphs (1 species), Marsupials (1 species) and Ungulates (1 species). Direct sequencing of PCR products demonstrated that specific mtSSU and mtLSU rRNA Pneumocystis sequence could be attributed to each mammalian species. No animal harbored P. carinii f. sp. hominis. Comparison of combined mtLSU and mtSSU aligned sequences confirmed cospeciation of P. carinii and corresponding mammalian hosts. P. carinii organisms isolated from mammals of the same zoological group systematically clustered together. Within each cluster, the genetic divergence between P. carinii organisms varied in terms of the phylogenetic divergence existing among the corresponding host species. However, the relative position of P. carinii groups (rodent, camivore or primate-derived P. carinii) could not be clearly determined. Further resolution will require the integration of additional sequence data. PMID- 11906017 TI - Cloning and sequence analysis of translational release factors eRF1 and eRF3 of Pneumocystis carinii. PMID- 11906018 TI - Detailed structure of Pneumocystis carinii chromosome ends. PMID- 11906019 TI - Typing of Pneumocystis carinii f.sp. hominis isolates from nasopharyngeal aspirates of immunocompetent infants with bronchiolitis by dihydropteroate synthase gene analysis. PMID- 11906020 TI - Pneumocystis carinii detection using nested-PCR in nasopharyngeal aspirates of immunocompetent infants with bronchiolitis. PMID- 11906021 TI - Typing of Pneumocystis carinii f.sp. hominis in patients with or without pneumocystosis. PMID- 11906022 TI - Distribution of DHPS mutations among ITS subtypes of P. carinii f. sp. hominis. PMID- 11906023 TI - Fatal spontaneous pneumocystosis in CD40 knockout mice. PMID- 11906024 TI - Stimulation of Pneumocystis carinii encystment following necropsy. PMID- 11906025 TI - A gene complex mediating cyst wall assembly and integrity in Pneumocystis carinii. PMID- 11906026 TI - Synaptonemal complexes in the pre-cyst of Pneumocystis carinii. PMID- 11906027 TI - Early acquisition of Pneumocystis carinii in neonatal rats using targeted PCR and oral swabs. PMID- 11906028 TI - Pneumocystis carinii dihydropteroate synthase genotypes in HIV-infected persons residing in San Francisco: possible implications for disease transmission. PMID- 11906029 TI - Putative transmissive form of Pneumocystis carinii f. sp. carinii. PMID- 11906030 TI - Free living amoebae as opportunistic hosts for intracellular bacterial parasites. PMID- 11906031 TI - Analysis of a pheromone receptor and MAP kinase suggest a sexual replicative cycle in Pneumocystis carinii. PMID- 11906032 TI - Definitive structural identities of 42 sterol components in Pneumocystis carinii. PMID- 11906033 TI - Pneumocystis carinii erg6 gene: sequencing and expression of recombinant SAM:sterol methyltransferase in heterologous systems. PMID- 11906034 TI - Pneumocystis carinii ITS typing: doubtful evidence of genotype-related virulence. PMID- 11906035 TI - PCR-RFLP analysis of the DHPS gene for the study of resistance of Pneumocystis carinii to sulpha drugs in patients with co-infection PCP/HIV. PMID- 11906036 TI - Urokinase plasminogen activator and TGF-beta production in immunosuppressed patients with and without Pneumocystis carinii. PMID- 11906037 TI - PCP occurring in HIV patients under HAART: assessment of specific immunity against P. carinii. PMID- 11906038 TI - Recombinant CD40 ligand administration does not decrease intensity of Pneumocystis carinii infection in scid mice. AB - X-linked Hyper IgM Syndrome (HIM) is a rare congenital immunodeficiency recently demonstrated to be caused by a mutation in the gene encoding CD40 ligand. These patients are susceptible to Pneumocystis carinii pneumonia, which implies an important role for CD40L in host defense against P. carinii. In this study we undertook to investigate whether treatment of P. carinii infected scid mice with murine recombinant CD40 ligand trimer (muCD40L) for 21 days would facilitate clearance of the organisms. We found no significant difference in organism burden in treated compared to control animals. Therefore in this model treatment with muCD40L alone is ineffective in clearing P. carinii infection. PMID- 11906039 TI - Mechanisms of amino acid and glucose uptake by Pneumocystis carinii. PMID- 11906040 TI - Pneumocystis carinii beta-glucan induces release of macrophage inflammatory protein-2 from primary rat alveolar epithelial cells via a receptor distinct from CD11b/CD18. PMID- 11906041 TI - Effect of the transcription factor GATA-2 on phagocytic activity of alveolar macrophages from Pneumocystis carinii-infected hosts. PMID- 11906042 TI - Cryptosporidium research 2001: workshop summary. PMID- 11906044 TI - Changes in alveolar macrophage number are early events in Pneumocystis carinii infection. PMID- 11906043 TI - Role of nuclear factor-kappa B in the activation of alveolar macrophages by fungal beta-glucans. PMID- 11906045 TI - Bronchoalveolar lavage fluid from Pneumocystis carinii-infected rats inhibits phagocytosis in normal alveolar macrophages. PMID- 11906046 TI - Production and role of nitric oxide in the alveolar immune response to Pneumocystis carinii. PMID- 11906048 TI - The interaction of atovaquone with the P. carinii cytochrome bc1 complex. PMID- 11906047 TI - Pneumocystis carinii does not induce maturation of human dendritic cells. PMID- 11906049 TI - Pneumocystis carinii synthesizes four ubiquinone homologs: inhibition by atovaquone and bupravaquone but not by stigmatellin. PMID- 11906050 TI - Toward rational design of species-specific inhibitors of Pneumocystis carinii thymidylate synthase and Toxoplasma gondii dihydrofolate reductase-thymidylate synthase. PMID- 11906051 TI - Development of an RT-PCR on the heat shock protein 70 gene for viability detection of Pneumocystis carinii f. sp. hominis in patients with pneumocystosis and in air sample. AB - To test the viability of Pneumocystis carinii f. sp. hominis, an RT-PCR assay that employs specific primers from the Heat Shock Protein 70 gene was developed. Using this method, the viability of P.c. hominis in bronchoalveolar lavage fluids from patients developing PCP and in the environment of PCP patients was established. PMID- 11906052 TI - Standardization of an in vitro drug screening assay by use of cryopreserved and characterized Pneumocystis carinii populations. PMID- 11906053 TI - Multilocus genotyping of Cryptosporidium isolates from human HIV-infected and animal hosts. PMID- 11906054 TI - Pneumocystis carinii dihydropteroate synthase mutations and treatment with sulfa or sulfone regimens: a proposal for standardized definitions for clinical evaluation. PMID- 11906055 TI - The pneumocystis genome project: update and issues. PMID- 11906056 TI - New nomenclature for the genus Pneumocystis. PMID- 11906057 TI - Toxoplasma gondii research: summary of the seventh international workshops on opportunistic protists. PMID- 11906058 TI - A multiplex PCR assay for molecular recognition of T. gondii stage-specific genes. PMID- 11906059 TI - Expression of toxoplasmic 65 kDa cystic mRNA by RT-PCR in patients with Toxoplasma gondii infection relapses. PMID- 11906061 TI - Removal of Toxoplasma gondii oocysts from sea water by eastern oysters (Crassostrea virginica). AB - Toxoplasma gondii infections have been reported in a number of marine mammals. Presently it is not known how these animals acquire T. gondii from their aquatic environment. The eastern oyster, Crassostrea virginica, has been shown to remove Cryptosporidiwn oocysts from seawater and a similar phenomenon may be occurring with T. gondii oocysts and marine invertebrates. The present study was done to determine if eastern oysters could remove and retain T. gondii oocysts from seawater. Oocysts of the VEG strain of T. gondii (1 x 10(6) oocysts) were placed in seawater (32 ppt NaCl) containing live eastern oysters. The infected seawater was removed one day postinoculation (PI) and replaced with fresh seawater. Selected oysters were removed at 1, 3 and 6 days PI. Hemolymph, gill washes, and oyster tissue were collected separately at each observation time. The oyster tissue was homogenized and all 3 samples fed separately to mice. Toxoplasma gondii positive mice were observed at each time period. The results indicate that T. gondii oocysts can be removed from seawater by eastern oysters and retain their infectivity. Contaminated raw oysters may serve as a source of T. gondii infection for marine mammals and humans. PMID- 11906060 TI - Production and characterization of a monoclonal antibody against nucleoside triphosphate hydrolase from Toxoplasma gondii. PMID- 11906062 TI - Effects of recent methyl mercury exposure on acute toxoplasmosis in CBA/J mice. PMID- 11906063 TI - A multilocus genotypic analysis of Cryptosporidium meleagridis. AB - Cryptosporidium meleagridis is a common cause of cryptosporidiosis in birds. In addition, recent reports have described the parasite as an etiologic agent of cryptosporidiosis in both immunocompetent and immunocompromised humans. Therefore, it is important to genetically characterize isolates of C. meleagridis from different hosts and geographic areas, and to develop molecular tools to differentiate isolates from various hosts or areas. In this study, a total of 11 isolates of Cryptosporidium meleagridis from both human and avian hosts were examined at three genetic loci: the small-subunit rRNA, 60-kDa glycoprotein precursor, and 70-kDa heat shock protein genes. Two genotypes of C. meleagridis were seen at the small-subunit rRNA locus. These differed from each other by the presence or lack of a heterogeneous copy of the gene and an ATT repeat. The 60 kDa glycoprotein precursor gene divided these eleven isolates of C. meleagridis into six genotypes with high sequence diversity between groups. The highest genetic heterogeneity, however, was seen at the 70-kDa heat shock protein locus, and was primarily present at the 3' end of the gene. This heterogeneity separated eight isolates of C. meleagridis into six genotypes. These data could be useful in the development of molecular tools to promote understanding of the transmission of C. meleagridis in humans. PMID- 11906064 TI - Stage-specific polyamine metabolism in Trypanosoma cruzi. PMID- 11906065 TI - Molecular tool's contribution to the detection of Cryptosporidium. PMID- 11906066 TI - A population genetic study of the Cryptosporidium parvum human genotype parasites. PMID- 11906067 TI - A comparison of Cryptosporidium subgenotypes from several geographic regions. PMID- 11906068 TI - Genetic characterization of Cryptosporidium parvum isolates from cattle in Portugal: animal and human implications. PMID- 11906069 TI - Seasonal shift in Cryptosporidium parvum transmission cycles in New Zealand. PMID- 11906070 TI - Involvement of insects in the dissemination of Cryptosporidium in the environment. PMID- 11906071 TI - Inactivation of Cryptosporidium parvum oocysts by bacterial strains. PMID- 11906072 TI - Current research on free-living amebae causing granulomatous amebic encephalitis. PMID- 11906073 TI - Immunohistochemistry based assay to determine the effects of treatments on Cryptosporidium parvum viability. AB - Cell culture infectivity assays can provide an accurate means of detecting viable Cryptosporidium parvum oocysts from environmental samples or to test the effects of various treatments on oocyst infectivity. Cell culture assays can also be used to test candidate chemotherapeutic agents. The use of a human cell line provides a situation close to human infection. The present assay uses an anti Cryptospordium primary antibody, combined with a biotinylated secondary antibody, and an immunoperoxidase detection system. Cryptosporidium parvum oocysts excysted in vitro when placed on monolayers of HCT-8 cells and developmental stages including schizonts and merozoites were visualized using light microscopy of the immunoperoxidase stained slides and by transmission electron microscopy of infected HCT-8 cell cultures. Because the immunoperoxidase system used gives a permanent preparation, the cell cultures can be retained and examined later. Dose titration of oocysts indicated that as few as 50 inoculated oocysts could be detected. The activity of paromomycin was evaluated in this system and 500 microg/ml produced a 97.8% reduction in infection. PMID- 11906074 TI - Microarray-based expression analysis of human epithelial cell response to Cryptosporidium parvum infection. PMID- 11906075 TI - Targeted disruption of CSL ligand-host cell receptor interaction in treatment of Cryptosporidium parvum infection. PMID- 11906076 TI - Microsporidia 2001: Cincinnati. PMID- 11906077 TI - Delta-9-tetrahydrocannabinol (THC), the major psychoactive component of marijuana, exacerbates brain infection by Acanthamoeba. PMID- 11906079 TI - A proteome approach to the host-parasite interaction of the microsporidian Encephalitozoon intestinalis. PMID- 11906078 TI - Inter-strain variability of insertion/deletion events in the Encephalitozoon cuniculi genome: a comparative KARD-PFGE analysis. AB - We applied a two-dimensional pulsed-field gel electrophoresis procedure to the genomes of two karyotype variants assigned to two different strains of the microsporidian Encephalitozoon cuniculi, termed D (strain III) and F (strain II). Data obtained for BssHII and MluI restriction fragment length polymorphisms in each chromosome are compiled and compared to the reference strain I variant A. Six Insertion/Deletion (InDels) are found in subterminal position, some of these being characteristic of either D or F. Like in strain 1, the terminal fragments extending between each telomere and rDNA locus are conserved in length for each chromosome. They are however smaller than in reference variant. This size reduction is estimated to be 2.5 kbp for the strain III isolate and 3.5 kbp for the strain II isolate. We hypothesize that for the three E. cuniculi strains, all chromosome extremities are prone to a constant process of sequence homogenization through mitotic recombination between conserved regions. PMID- 11906080 TI - The identification of rRNA maturation sites in the microsporidian Encephalitozoon cuniculi argues against the full excision of presumed ITS1 sequence. AB - In Encephalitozoon cuniculi like in other microsporidia, the primary transcript for SSU and LSU rRNAs includes only one internal transcribed spacer (ITS1) which separates SSU rRNA from the 5.8S region associated with LSU rRNA. The extraction of total RNA from E. cuniculi-infected MRC5 cells using a hot phenol/chloroform procedure enabled us to perform primer extension and S1 nuclease protection experiments in the aim of identifying rRNA maturation sites. Our data support a simple processing (four cleavage sites) with elimination of only nine nucleotides between SSU and LSU rRNA regions. Most of the presumed ITS1 sequence characterized by strain-dependent polymorphism therefore remains linked to SSU rRNA 3' end. A new secondary structure for the sixth domain of E. cuniculi LSU rRNA is proposed following the identification of its 3' terminus. PMID- 11906081 TI - Genotyping Encephalitozoon parasites using multilocus analyses of genes with repetitive sequences. PMID- 11906082 TI - Genetic and immunologic characterization of seven Encephalitozoon hellem human strains. PMID- 11906083 TI - Methodology of the diagnosis of microsporidiosis in urine and pulmonary specimens from AIDS patients. PMID- 11906084 TI - Balamuthia mandrillaris: its pathogenic potential. PMID- 11906085 TI - Monoclonal antibody enabling the diagnosis of Encephalitozoon intestinalis in fecal specimens: importance of the mode of selection of hybridomas. PMID- 11906086 TI - An ELISA test to detect human serum antibodies reactive with Encephalitozoon intestinalis. PMID- 11906087 TI - Seroprevalence of anti-Encephalitozoon antibodies in Spanish immunocompetent subjects. PMID- 11906089 TI - Brachiola algerae sporoplasms. PMID- 11906088 TI - Encephalitozoon cuniculi infection in mice with the chronic granulomatous disease (CGD) disorder. PMID- 11906090 TI - Phagocytosis of Nosema grylli (Microsporida, Nosematidae) spores in vivo and in vitro. PMID- 11906091 TI - Development of Thelohania solenopsae in red imported fire ants Solenopsis invicta from polygynous colonies results in formation of three spore types. PMID- 11906092 TI - Visualization of early golgi compartments at proliferate and sporogenic stages of a microsporidian Nosema grylli. PMID- 11906093 TI - Microsporidian methionine aminopeptidase type 2. PMID- 11906094 TI - Effects of gamma irradiation on the survival of Encephalitozoon intestinalis spores. PMID- 11906095 TI - SL-11158, a synthetic oligoamine, inhibits polyamine metabolism of Encephalitozoon cuniculi. PMID- 11906096 TI - In vitro and in vivo evaluation of aminopeptidase inhibitors as antimicrosporidial therapies. PMID- 11906097 TI - In vitro activity of antimitotic compounds against the microsporidium Encephalitozoon intestinalis. PMID- 11906098 TI - Nature's revelation: seed cells of the body--stem cells. PMID- 11906099 TI - Citrus tissue culture employing vegetative explants. AB - Citrus being a number one fruit of the world due to its high nutritional value, huge production of fruits and fruit products, the citrus industry may be considered a major fruit industry. Though citrus orchard area in India is comparable to USA, the produce is far less, while its export is nil. Biotechnology has played an outstanding role in boosting the citrus industry, e.g., in Spain, which is now the biggest exporter of citrus fruit with the application of micrografting. Amongst the fruit trees, perhaps the maximum tissue culture research has been done in citrus during the past four decades, however, the results of practical value are meagre. The shortfalls in citrus tissue culture research and some advancements made in this direction along with bright prospects are highlighted, restricting the review to vegetative explants only. Whilst utilization of nucellar embryogenesis is limited to rootstocks, the other aspects, like, regeneration and proliferation of shoot meristems measuring 200 microm in length--a global breakthrough--of two commercially important scion species, Citrus aurantifolia and C. sinensis and an important rootstock, C. limonia, improvement of micrografting technique, cloning of the same two scion species as well as some Indian rootstock species, employing nodal stem segments of mature trees, of immense practical value have been elaborated. A rare phenomenon of shift in the morphogenetic pattern of differentiation from shoot bud differentiation to embryoid formation occurred during the long-term culture of stem callus of C. grandis. Stem callus-regenerated plants of C. aurantifolia, C. sinensis and C. grandis showed variation in their ploidy levels and a somaclonal variant of C. sinensis, which produced seedless fruits was isolated. Tailoring of rooting in microshoots to a tap root-like system by changing the inorganic salt composition of the rooting medium, resulting in 100% transplant success, and germplasm preservation through normal growth culture of shoots of C grandis without loss of regeneration capacity during 31 years, observed so far, are some other significant results. Plants of C. aurantifolia and C. sinensis raised from shoot meristem and micrografting were grown in a nethouse and those from nodal stem segments in the field along with the in vitro-raised plants of rootstocks, namely, C. jambhiri, C. karna and C. limonia. All the plants showed normal healthy growth. Significantly enough, the meristem regenerated plants of C. aurantifolia attained the reproductive phase just in 1 year of transplantation to soil similar to those raised from nodal stem segments of mature trees, which also produced normal fruits in the subsequent year while growing under field conditions. Thus, a significant fundamental concept of a maturity factor, carried over through as small a shoot meristem as 200 microm in length to cloned plants has been demonstrated. The concept is of far-reaching significance in citrus industry besides production of pathogen-free orchards. PMID- 11906100 TI - Effect of estradiol-17beta on cell area, lumen area and trehalase activity of posterior silk gland of Bombyx mori L. AB - Estradiol-17beta (E2) at the dose of 1 microg/g caused an increase in cell area, lumen area and the total (cell + lumen) area of posterior silk gland (PSG) in Bombyx mori indicating that exogenously applied estradiol-17beta has a regulatory influence on silk gland activity. A dose-dependent variation in trehalase activity of PSG was found on the 5th day after topical administration of estradiol on 1st and 2nd day of the fifth larval instar. Of all the doses of E2 used, 1 microg/g dose had maximum stimulatory effect on trehalase activity. Co administration of each of a specific receptor antagonist for estradiol, the ICI 182780 and a protein biosynthetic blocker, cycloheximide with E2 suppressed the E2-induced increase in silk gland activity. The results suggest some specific metabolic action of E2 on silk gland and offer a promising way for future investigations regarding the physiological significance of vertebrate steroids in insects. PMID- 11906101 TI - Specific limb abnormalities induced by hydrogen peroxide in tadpoles of Indian jumping frog, Polypedates maculatus. AB - Hydrogen peroxide (H2O2), one of the reactive oxygen intermediates (ROI) and a potential inducer of nuclear transcription factors induces consistent type of abnormal limb development (truncated with bent skeletal elements) in the tadpoles of Indian jumping frog, Polypedates maculatus. PMID- 11906102 TI - Subacute toxicity of anilofos, a new organophosphorus herbicide, in male rats: effect on some physical attributes and acetylcholinesterase activity. AB - In acute toxicity study, rats showed dose-dependent signs of cholinergic hyperactivity and behavioural alterations. Maximum intensity of symptoms was not associated with mortality. Oral LD50 was 1681 mg/kg. In subacute toxicity study, rats were orally administered 50, 100 or 200 mg/kg of anilofos once daily for 28 days. Signs and symptoms were observed mainly with 200mg/kg. At this dose, anilofos induced hypothermia and progressive weight loss. None of the anilofos treated rats died. Weight of brain, lung, testis was not altered, while of liver, heart, spleen and kidney increased. Anilofos inhibited cholinesterase (ChE) activities of erythrocyte (41-67%), plasma (36%), blood (37-64%), brain (63-73%) and liver (28-48%). Total protein was decreased in plasma and liver. Results indicate moderate toxic potential of anilofos in mammals, substantial contribution of CNS-mediated effects in causing anilofos toxicity and no direct relationship between hypothermia and level of ChE inhibition. PMID- 11906103 TI - Subacute toxicity of anilofos, a new organophosphorus herbicide in male rats: effect on lipid peroxidation and ATPase activity. AB - Effects of anilofos on lipid peroxidation--an index of oxidative stress, ATPase activity--an integral part of active transport mechanisms for cations, GSH level and GST activity were evaluated in blood (erythrocyte/plasma), brain and liver of male rats after daily oral exposure to 50, 100 or 200 mg/kg for 28 days. None of the doses increased lipid peroxidation. The lowest dose, rather, produced marginally significant decrease in peroxidation in liver. Different doses of anilofos decreased GSH content and activities of GST and ATPases. Inhibition of total ATPase (34-44%) and Na+-K+-ATPase (45-52%) activities was maximum in liver, while that of Mg2+-ATPase (46-56%) was more in erythrocyte. Results indicate that anilofos may not cause oxidative damage to cell membrane in repeatedly exposed animals and may cause neuronal/cellular dysfunction by affecting ionic transport across cell membrane. PMID- 11906104 TI - Surfactant-induced lipid peroxidation in a tropical euryhaline teleost Oreochromis mossambicus (Tilapia) adapted to fresh water. AB - Exposure to anionic (sodium dodecyl sulfate, SDS), cationic (cetyl trimethyl ammonium bromide CTAB) and non ionic (Triton X-100) surfactants at a sub lethal concentration of 1 ppm resulted in severe oxidative stress in the hepatic, renal and cardiac tissues of fresh water adapted Oreochromis mossambicus. Hepatic catalase showed significant increase (P<0.001) in all the surfactant exposed fish, but the renal enzyme was significantly increased only in CTAB dosed fish (P<0.001) and the cardiac enzyme showed significant increase in Triton (P<0.05) and CTAB dosed fish (P<0.001). SOD levels were significantly increased (P<0.001) in hepatic, renal and cardiac tissues of all the surfactant-treated fish. Glutathione reductase also was significantly increased (P<0.001) in the hepatic and renal tissues of surfactant dosed fish except cardiac tissues of CTAB exposed animals. Glutathione levels in the tissues studied were significantly higher in the surfactant treated animals (P<0.001) whereas malondialdehyde levels were significantly elevated only in the hepatic tissues of animals exposed to Triton (P<0.001). The surfactants based on their charge, antioxidant profile and in vivo metabolism may be arranged in the order of decreasing toxicity as CTAB > Triton > SDS. Thus it may be inferred from the present study that the antioxidant defenses and the in vivo metabolism of the surfactants are key factors in deciding the surfactant toxicity. PMID- 11906105 TI - Changes in haemolymph constituents of American bollworm, Helicoverpa armigera (Hubner), infected with nucleopolyhedrovirus. AB - Six types of haemocytes viz., prohaemocytes, plasmatocytes (round, fusiform, vermiform and spindle shaped), granular cells, spherule cells, oenocytoids and adipohaemocytes were found in the haemolymph of larvae of American bollworm H. armigera. The total and differential haemocyte counts (THC and DHC) in H. armigera haemolymph were affected by nucleopolyhedrovirus (NPV) treatment. There was a general decrease in THC in response to NPV treatment in both young and old larvae. However the decrease was more apparent in 5 and 8 day old larvae than in 10 day old larvae. The differential haemocytes showed less of granular cells and more of spherule cells and prohaemocytes in the old larvae. Plasmatocytes and granular cells in 10 day old larvae initially phagocytosed polyhedra; however, disintegrated after 3 to 4 hr. The haemolymph of NPV treated larvae melanized slowly particularly in old larvae. Phenoloxidase (PO) activity decreased positively with granular cells and oenocytoids in 10 day old treated larvae. Cellular fraction had high level of PO activity, which was transferred to plasma in response to NPV infection in the older larvae. The role of NPV pathogenesis vis-a-vis immunity in insect is discussed. PMID- 11906106 TI - Effect of sublethal concentration of Bacillus thuringiensis var. kurstaki on food and developmental needs of the american bollworm, Helicoverpa armigera (Hubner). AB - Effect of sublethal concentration of B. thuringiensis on the first, third, fourth and fifth instar larvae of the American bollworm, H. armigera was investigated to study their response to food consumption, digestion, utilization, and their development till adult formation. The young larvae surviving B. thuringiensis treatment in their first instar and third instar delayed larval period by two to three days, but did not consume more food as compared to control. However, they showed higher digestibility of food as compared to control, which was compensated by their reduced ability to utilize the digested food for body substance. Contrary to the effect on first and third instar larvae, the fifth instar larvae surviving B. thuringiensis treatment in its fourth instar consumed less food, showed less absorption efficiency in digesting food, but compensated by increase in the utilization of ingested and digested food into body substance. Insects surviving B. thuringiensis HD-1 sublethal toxicity adapted to normal larval growth when fed on untreated food, depending upon insect growth prior to treatment. The moths emerging from B.thuringiensis treated larvae had sex ratio favouring females, and adult pairs laid less fertile eggs than those from the untreated ones. The response of B. thuringiensis treated larvae to their food and developmental needs is discussed. PMID- 11906107 TI - Optimisation of fermentation conditions for gluconic acid production by a mutant of Aspergillus niger. AB - Aspergillus niger ORS-4, isolated from the sugarcane industry waste materials was found to produce notable level of gluconic acid. From this strain, a mutant Aspergillus niger ORS-4.410 having remarkable increase in gluconic acid production was isolated and compared for fermentation properties. Among the various substrates used, glucose resulted into maximum production of gluconic acid (78.04 g/L). 12% concentration led to maximum production. Effect of spore age and inoculum level on fermentation indicated an inoculum level of 2% of the 4 7 days old spores were best suited for gluconic acid production. Maximum gluconate production could be achieved after 10-12 days of the fermentation at 30 degrees C and at a pH of 5.5. Kinetic analysis of production indicated that growth of the mutant was favoured during initial stages of the fermentation (4-8 days) and production increased during the subsequent 8-12 days of the fermentation. CaCO3 and varying concentrations of different nutrients affected the production of gluconic acid. Analysis of variance for the factors evaluated the significant difference in the production levels. PMID- 11906108 TI - Optimization of some additives to improve protease production under SSF. AB - In a locally isolated Rhizopus oryzae strain highest-production of protease (388.54/g wheat bran) was observed in presence of Tween-80 and dioctyl sodium sulfosuccinate individually at 40mg/g wheat bran concentration. Under solid state fermentation biotin (0.0025mg/g wheat bran); Ca2+ (0.05mg/g wheat bran) and 1 Naphthyl acetic acid (0.01mg/g wheat bran) also showed some inducing effect on the synthesis of the enzyme protease by solid state fermentation. PMID- 11906109 TI - Distinguishing clonal apple rootstocks by isozymes banding patterns. AB - Molecular characterisation of clonal apple rootstocks using isozymes was carried out to identify isozyme polymorphism in seven clonal apple rootstocks and to identify the most characteristic and stable enzyme markers for each individual rootstock. Five enzyme systems were studied out of which polyphenol oxidase, malate dehydrogenase, acid phosphatase and peroxidase were useful in discriminating among the rootstocks. The peroxidase enzyme system showed maximum variation and esterase showed the least variation among the rootstocks. Out of seven rootstocks, three were distinguished on the basis of one enzyme system only (M.3 with MDH or PER, M.7 with PPO or PER and MM. 111 with MDH). Out of the sixteen loci studied seven were found to be polymorphic. Genetic variation among the rootstocks was explained on the basis of various parameters. The percentage of polymorphic loci varied from 13.33 to 35.71 per cent. PMID- 11906110 TI - Activity of glutathione related enzymes and ovarian steroid hormones in different sizes of follicles from goat and sheep ovary of different reproductive stages. AB - The investigations on enzymes related to glutathione like glutathione-S transferase (GST) and glutathione peroxidase (GSH-Px) have been carried out mostly in human and rat ovaries, however the studies on these enzymes in ruminants are relatively absent. In the present study the changes in the activity of these enzymes, in different sizes of follicles from goat and sheep ovaries of different reproductive stages, were investigated. The results demonstrated that the activity of the enzyme GST increased with the increase in size of the follicles from small to large follicles of follicular phase ovary and from small to medium follicles of luteal phase ovary in both the species, thereafter it decreased in large follicles of luteal phase ovary. There was increasing pattern in the activity of GSH-Px in the follicular phase follicles and a decreasing pattern in the luteal phase follicles from both the species. Thus the changes in the activity of glutathione related enzymes namely GST and GSH-Px in different size follicles from both the species during different reproductive phases are evident from the results. It is reasonable, therefore, to assume that these enzymes may have functional role in the steroid hormone metabolism in ruminant ovary as reported in human ovary. PMID- 11906111 TI - Effect of metronidazole on spermatogenesis and FSH, LH and testosterone levels of pre-pubertal rats. AB - Metronidazole, a 5-nitroimidazole drug has been reported to decrease testicular weight, testicular and epididymal spermatid counts and causes abnormal sperm morphology with degeneration of seminiferous tubules with 6 weeks treatment of metronidazole (400 mg/kg, day). In contrast to DNA flow cytometry (FCM), the histological and gravimetric parameters do not allow a rapid, sensitive, objective and multiparameteric evaluation of reproductive toxicants on spermatogenesis. Moreover, the exact mechanisms for such an effect are not entirely clear. The present study was therefore undertaken to assess the effects of intraperitoneal (i.p.) administration of metronidazole 400 mg/kg daily for 30 days on testicular germ cell changes assessed by DNA (FCM) and hormone levels of testosterone, FSH and LH in pre-pubertal rats. A significant reduction in the haploid cell population in metronidazole treated groups as compared to saline treated controls was observed. The mean serum FSH, LH and testosterone value were also lowered in treated animals. Thus, the spermatotoxic effects of metronidazole were probably mediated by decrease in the circulating hormones responsible for spermatogenesis. PMID- 11906112 TI - Effect of dexamethasone on implantation and pregnancy in albino rats. AB - Administration of 3 mg/kg body weight of dexamethasone from day 1 or 3 to 7 of pregnancy did not prevent implantation in albino rats. But the same dose when administered from day 8 to 11 resulted in complete abortion / resorption in all rats. Administration of 2 mg / kg body weight of dexamethasone from day 8 to 11 of pregnancy held no effect on the foetal survival. The results indicate that a high dose of dexamethasone does not affect implantation but the same dose affects the more advanced stages of pregnancy. PMID- 11906113 TI - Antioxidant and anti-inflammatory activities of methanol extract of Phellinus rimosus (Berk) Pilat. AB - The methanolic extract of a macrofungus, P. rimosus possessed significant in vitro superoxide anion, hydroxyl radical and nitric oxide scavenging and lipid peroxidation inhibiting activities. The anti-inflammatory activity of the extract was evaluated in carrageenan and dextran induced acute and formalin induced chronic inflammatory models in mice. The extract showed remarkable anti inflammatory activity in both models, comparable to the standard reference drug diclofenac. The results suggest that the anti-inflammatory activity of the methanol extract of P. rimosus is possibly attributed to it's free radical scavenging properties. The findings also reveal the potential therapeutic value of P.rimosus extract as an antiinflammatory agent. PMID- 11906114 TI - Influence of zinc on cardiac and serum biochemical parameters in rabbits. AB - The pattern of lipid profiles and organic constituents of cardiac and serum tissues of rabbits were studied on treatment with cholesterol, zinc and zinc + cholesterol. Total carbohydrate and total protein levels were decreased with elevated lipid levels in cholesterol fed rabbits. However, the zinc and cholesterol + zinc fed rabbits showed decreased lipid fractions in cardiac and serum tissues leading to reduced atherosclerotic process in rabbits. These results suggest that the zinc is acting as a hypolipidaemic and anti atherogenic agent in experimental rabbits. PMID- 11906115 TI - Effect of brahma rasayana on antioxidant system after radiation. AB - Oral administration of brahma rasayana (BR; 50 mg/animal for 10 and 30 days) significantly increased the liver antioxidant enzymes such as superoxide dismutase (SOD), catalase(CAT) and tissue and serum levels of reduced glutathione (GSH). Whole body irradiation suppressed the levels of SOD, CAT and GSH. Reduced activity of SOD, CAT and GSH was significantly elevated by treatment with BR after radiation treatment. Similarly radiation exposure induced increase in serum and liver lipid peroxides was significantly reduced by further treatment with BR. The results indicate that BR could ameliorate the oxidative damage produced in the body by radiation and may be useful as an adjuvant during radiation therapy. PMID- 11906116 TI - Antioxidant property of Smilex china Linn. AB - Smilex china Linn. (Smilacaceae Syn Liliaceae) has special pharmacognostic feature as its root is multiseriate, with sclerenchymatic pericycle. The alcoholic extract of rhizome of S. china shows significant protection against FeSO4 induced lipid peroxidation in rat liver homogenate, but has no effect on the rate of oxidation of reduced glutathione. This fraction scavenges the superoxide and hydroxyl radicals, but the effect was more towards the removal of superoxide than that of hydroxyl radicals. Thus it could be concluded that rhizome of S. china has strong antioxidant property. PMID- 11906117 TI - Biochemical evaluation of lipid and oxidative stress status in relation to high fat-high antioxidant diets. AB - Free radicals are produced through biological processes and environmental interactions. They are metabolised by the enzymatic and non-enzymatic antioxidants present in the tissues. In this study, a 90 days long feeding of high fat diet to rats, resulted in significantly elevating the lipid and oxidative stress levels of the rat liver and blood as became evident from the changes in the levels of lipids, thiobarbituric acid reactive substances(TBARS), reduced glutathione (GSH), and three hepatic antioxidant enzymes; glutathione peroxidase (EC 1.11.1.9), catalase (EC 1.11.1.6) and superoxide dismuatase (EC 1.15.1.1). However, a concomitant feeding of high antioxidant combination, as high fat high antioxidant diets, reduced the lipid levels and diminished the oxidative stress. The results suggest that apart from reducing lipid levels, dietary antioxidants also support endogenous antioxidants in their oxidative stress reducing endeavours. PMID- 11906118 TI - High frequency in vitro propagation of Phyllanthus amarus Schum. & Thom. by shoot tip culture. AB - With the aim of micropropagation of Phyllanthus amarus, an important medicinal herb, shoot tips were cultured in Murashige and Skoog's medium supplemented with kinetin/ BAP singly or in combination with IAA. Growth regulators at lower range (0.1-1.0 mg L(-1)) stimulated direct regeneration of shoots. Kinetin was superior to BAP and kinetin-IAA combination was more suitable than kinetin alone. About 15 shoots were yielded per explant after 30 days of culture in the medium containing kinetin and IAA both at 0.1mg L(-1). The cluster of proliferated shoots elongated and rooted simultaneously under the same treatment following another subculture, thus shortening the total time schedule of micropropagation. Shoot tips of regenerated shoots were continuously used to regenerate new shoots with periodic transfer to fresh medium resulting in a steady supply of normal, healthy plants without any deviation in the production rate during a continuous one year culture. Micropropagated plants were successfully established in soil with high survivality (80%). PMID- 11906119 TI - In vitro propagation of Mussaenda erythrophylla Schum and Thom cv. scarlet through multiple shoot regeneration. AB - Axillary bud explants were induced to form shoots on Murashige and Skoog's (MS)' basal medium. Best yield (9 shoots per explant) was obtained when the medium was supplemented with adenine sulphate (40 mg/L) and 6-benzylaminopurine (2.25 mg/L). The shoots were rooted on half strength MS basal medium supplemented with indole butyric acid (0.5 mg/L) and having thiamin-HCl (800 mg/L). Regenerated plantlets were successfully acclimatized. This is the first report of micropropagation in the genus Mussaenda without callus intervention. PMID- 11906120 TI - Changes in rubisco content in relation to saccharides accumulation in wheat leaves during day. AB - Two wheat varieties, T. durum (HD 4502) and T. aestivum (Kalyansona) were examined for photosynthesis rate and contents of sugars and rubisco protein in the flag leaf, at forenoon and afternoon at anthesis stage. A decrease in photosynthesis rate was observed in the afternoon compared to forenoon in both the varieties and was associated with an increase in non-reducing sugars and a decrease in rubisco content in the leaves. PMID- 11906121 TI - Allelopathic effects of Lantana camara Linn. on spore germination of Asterella angusta Steph.--a liverwort. AB - Extract from root, stem and leaf of L. camara proved inhibitory for germination of the spores of A. angusta. Leaf extract was found to exhibit maximum allelopathic potentiality followed by stem and root extract and may be interpreted to be the result of phytotoxic substances which are possibly synthesized in the leaf and translocated to other organs. PMID- 11906122 TI - Evaluation of immunoglobulin G and its subclasses in aplastic anemia. AB - We selected 31 patients with aplastic anemia to measure their total immunoglobulin (Ig) G and IgG subclasses and compared the results obtained with those obtained in the normal control group (n=461). Patients with aplastic anemia were classified into three subgroups according to severity: mild subgroup (n=14), moderate subgroup (n=12) and severe subgroup (n=5). The decrease in total IgG was recognized in all these three subgroups (p<0.001). Although IgG1 decreased in severe subgroup (p<0.01), it showed no significant difference in mild and moderate subgroups. The decrease in IgG2 and IgG3 was observed in all these three subgroups (p<0.001 and p<0.01, respectively). Although IgG4 decreased in mild subgroup (p<0.05), it showed no significant difference in moderate and severe subgroups. We speculated that the measurement of IgG subclasses was useful in understanding the pathophysiology of humoral immunity in aplastic anemia. PMID- 11906123 TI - Clinical studies on the visualization of gastric lesions using virtual CT endoscopy. AB - OBJECTIVE: We evaluated the clinical usefulness of virtual CT gastroendoscopy (VCTGE). METHODS: The subjects were 124 patients with endoscopically identified gastric lesions. VCTGE images were obtained using a GE-Navigator. We evaluated VCTGE in the visualization of gastric lesions for their presence and morphology. RESULTS: The detection rate of all gastric lesions by VCTGE was 76% (94 of 124 patients). The smallest detectable early gastric cancer was II c measuring 10 x 8 mm. The detection rates of each gastric lesion by VCTGE were 73% in early gastric cancer, and 90% in advanced gastric cancer. VCTGE imaging in the advanced gastric cancer was good in 12 (30%), fair in 25 (60%) and poor in 4 (10%). VCTGE imaging in early gastric cancer was good in 20 (46%), fair in 12 (27%) and poor in 12 (27%). The significance P level was 0.005 between the evaluation of the imaging of advanced and early gastric cancer. CONCLUSIONS: VCTGE visualized the characteristics of diverse gastric lesions and was considered useful for the detection and the diagnosis of these lesions. PMID- 11906124 TI - Fast MR imaging and ultrafast MR imaging of fetal central nervous system abnormalities. AB - The aims of this study were two: (1) to compare the efficacy of fast MRI (breath hold fast spin-echo T2-weighted and fast gradient-echo T1-weighted sequence) and ultrafast MRI (half-Fourier acquisition single-shot turbo spin-echo sequence) in evaluation of fetal central nervous system (CNS) abnormalities at late gestational age, and (2) to compare the capability of fast MRI and ultrafast MRI to assess fetal CNS abnormalities with that of prenatal ultrasonography (US). Forty-nine women with fetuses at gestational ages of 26-39 weeks underwent fast MRI (29 patients) or ultrafast MRI (20 patients). In detection of motion artifact, visualization of the lateral and 4th ventricles, and differentiation between gray and white matter in cerebral hemispheres, ultrafast MRI was significantly superior to fast MRI (p< 0.0001, Mann-Whitney U test). In 25 of 43 cases, US and MR diagnoses were the same and consistent with postnatal diagnosis. In 10 of 43 cases, MRI demonstrated findings additional to or different from those of US, and MR findings were confirmed postnatally. MRI, particularly ultrafast MRI, is useful for demonstrating CNS abnormalities in situations in which US is suggestive but not definitive. PMID- 11906125 TI - Installation and experience of an automatic scheduling system for multiple diagnostic examinations: in search of maximum utilization of regional health care resources. AB - We installed a scheduling system that optimally schedules multiple appointments for various diagnostic examinations for a patient based on patient characteristics, disease characteristics and conditions, characteristics of diagnostic examinations, possible interactions between two successive examinations, and features and availability of diagnostic equipment. The system consists of four client terminals, 12 laboratory terminals, and one server. After the run-in period, the system started operation in July 2000. A total of 14353 examinations involving 11447 patients were managed over 7 months. On average, approximately 82 patients per day underwent approximately 103 examinations. On average, 16.1 patients a day requested 2 or more examinations in a scheduling session (approx. 2.3 examinations/patient; maximum 5 examinations for a single patient). After reading the request sheet(s) (OCR sheet(s)), suggested time/date slots of examination(s) were displayed on the client terminal within 10 seconds. The average time required for a patient to establish his or her schedule was approximately 2 minutes. Thus, the system greatly mitigated the load on health care professionals in scheduling appointments for examinations. In general, patients had a positive impression of the system. Furthermore, networking through these health care facilities and implementing this automatic scheduling system as a centralized appointment system can easily establish a "virtual diagnostic examination center" that fully utilizes the diagnostic equipment and staff available at community health care facilities. We estimate that approximately 30% of the diagnostic examinations in a health care facility can be referred to another facility in this cooperative system. PMID- 11906126 TI - Bilirubin as a tracer for detecting gastroesophageal reflux. AB - PURPOSE: The usefulness of intraesophageal monitoring of bilirubin, in diagnosing gastroesophageal reflux, was studied. PATIENTS AND METHODS: Bilirubin concentration and pH at 5 cm oral and anal to the esophagogastric junction were monitored for 24 hours in 19 patients with reflux esophagitis. The duration of bilirubin presented and pH less than 4.0 were obtained as the holding time (HT) of bilirubin and acid, respectively. RESULTS: There was no difference between HT of bilirubin and acid in the stomach and esophagus. In the stomach, HT of bilirubin did not correlate with acid. Whereas, in both of bilirubin and acid, HT in the esophagus correlated significantly with that in the stomach. The correlation was more definite for bilirubin than acid. CONCLUSIONS: Bilirubin and acid presented in the upper stomach for sufficient period independently. Bilirubin monitoring was useful to evaluate the etiology of damaging the esophageal mucosa and causes of symptoms by estimating duodenogastro-esophageal reflux, which has synergistic effect with acid. PMID- 11906127 TI - Three-dimensional gadolinium-enhanced MR angiography of the intracranial venous system. AB - PURPOSE: The purpose of this study was to assess the usefulness of three dimensional (3D) gadolinium-enhanced magnetic resonance (MR) angiography for evaluation of the intracranial venous system. MATERIALS AND METHODS: Thirty-three patients underwent 3D dynamic gadolinium-enhanced MR angiography as well as two dimensional (2D) time-of-flight (TOF) MR venography in transverse and coronal planes and conventional catheter cerebral angiography with digital subtraction. MR venography was displayed using a maximum-intensity-projection (MIP) algorithm. The acquisition time of 3D gadolinium-enhanced MR angiography was 102 seconds, and that of 2D TOF MR venography was about 7 minutes in the transverse plane and about 9 minutes in the coronal plane. Degree of visualization of the intracranial venous system on each MR sequence was compared with that on conventional catheter cerebral angiography as a standard. RESULTS: Superficial cortical veins and the dural sinus were better visualized on 3D gadolinium-enhanced MR angiography than on 2D TOF MR venography. CONCLUSION: Three-dimensional gadolinium-enhanced MR angiography is noninvasive and very useful for imaging of the intracranial venous system. It can replace 2D TOF MR venography not only because of its short examination time but because it better demonstrates intracranial venous structures. For evaluation of the SSS, lateral sinus, sigmoid sinus and straight sinus in particular, conventional catheter angiography seems to be unnecessary. PMID- 11906128 TI - A centrifuge-less plasma separation method from whole blood anticoagulated with EDTA-2K for the use of clinical laboratory tests. AB - In the modern medical laboratory system, simple and rapid processing of specimens are required. In the current system with the transportation line, its centrifugation part would disturb smooth flow of the testing because it needs much time for the centrifugation. To solve the problems, a serum separation method was tried for the whole blood specimen using poly-L-lysine, concanavalin A and phyto-hemoagglutinin. Ploy-L-lysine with molecular weight 130,000 to 210,000 in a final concentration of 0.1% could accelerate blood sedimentation, although its supernatant contaminated platelets. Concanavalin and phytohemoagulutinin could accelerate the sedimentation and obtained plasma, but the method could yield enough amount of supernatant by 1 hour standing. As the purpose of this study is to develop a centrifugeless method, a sieve method using a steel mesh and a magnet was applied to the mixture of EDTA blood, red-cell adhesives and thrombin. The method was unique to separate plasma, but the yield was not so high and chemistry data were not fitted with serum data in some of tests. Thus, the trial would be a new technology, but it was judged that some further improvement will be needed technically. PMID- 11906130 TI - Clinical study on 53 cases of cadaveric kidney transplantation at Osaka City University Hospital. AB - BACKGROUND: In this study, we examined the effects of nonimmunological factors on cadaver kidney transplant. METHODS: Fifty-three cadaver kidney transplant recipients were studied. They were divided into two groups, Group I ( donor age<60, n=38 ) and Group II ( donor age > or = 60, n=15). These patients were studied to determine whether donor age and recipient Body Mass Index (BMI) affected transplant outcome. RESULTS: The 3 and 5-year overall graft / patient survival was 80.6 / 100% and 72.7 97.3%, respectively. The best S-Cr after transplantation was significantly (p<0.05) lower in Group I compared to Group II. The 1, 5 and 8-year graft survival was significantly (p<0.05) better in Group I. In the low BMI patients of Group II, 5-year graft survival was significantly (p<0.01) better than high BMI patients. DISCUSSION: An aged cadaver donor was a risk factor for decreased posttransplant renal functions and lower graft survival. However, if a recipient with a low BMI can be selected, the outcome of cadaver kidney transplants from aged donors may be improved. PMID- 11906129 TI - Large focal nodular hyperplasia of the liver: possible to evade surgical resection. AB - PURPOSE: Although recent advances in diagnostic imaging have allowed a number of patients with focal nodular hyperplasia (FNH) to avoid surgical treatment, the natural course of large FNH is still unknown. CASE REPORT: A 25-year-old man was admitted because of a large hepatic mass detected on routine examination in June 1998. The only laboratory abnormality was an elevated gamma-GTP. Computed tomography, angiography, positron emission tomography using F-18 fluorodeoxyglucose (FDG-PET), and scintigraphy using technetium-99m galactosylneoglycoalbumin(99mTc-NGA) demonstrated a spoke-wheel appearance of vessels, normal hepatocytes, and no malignancy. Histologic findings on needle biopsy were consistent with FNH. After informed consent, the patient agreed to observation. Two years after the initial diagnosis, he has no symptoms, and there are no changes in the size or character of the lesion on computed tomography. DISCUSSION: Careful observation of patients with FNH is required because its natural course is unknown and these lesions can bleed or rupture. PMID- 11906131 TI - The development of the plutonium lung clearance model for exposure estimation of the Mayak production association, nuclear plant workers. AB - The purpose of this study was to develop a biokinetic model that uses urinary plutonium excretion rate data to estimate the plutonium accumulation in the human respiratory tract after occupational exposure. The model is based on autopsy and urinalysis data, specifically the plutonium distribution between the respiratory tract and the remainder of the body, taken from 543 former workers of a radiochemical facility at the Mayak Production Association (MPA) plant. The metabolism of plutonium was represented with a compartmental model, which considers individual exposure histories and the inherent solubility properties of industrial plutonium aerosols. The transport properties of plutonium-containing aerosols were estimated by experimentally defining their in vitro solubility. The in vitro solubilities were found by dialysis in a Ringer's solution. Analysis of the autopsy data indicated that a considerable fraction of the inhaled plutonium is systemically redistributed rapidly after inhalation. After the initial dynamic period, a three-compartment model describes the retention in the respiratory tract. One compartment describes the nuclide retained in the lungs, the second compartment describes a plutonium lung concentration that exponentially decreases with time, and the third compartment describes the concentration in the pulmonary lymph nodes. The model parameters were estimated by minimizing sum squared of the error between the tissue and bioassay data and the model results. The parameters reflect the inverse relationship between plutonium retention in lungs and the experimentally derived aerosol transportability. The model was validated by comparing the autopsy results with in vivo data for 347 cases. The validation indicates that the model parameters are unbiased. This model is being used to estimate individual levels of nuclide accumulation and to compute radiation doses based upon the urinary excretion rates. PMID- 11906132 TI - Extrapulmonary organ distribution of plutonium in healthy workers exposed by chronic inhalation at the Mayak production association. AB - The systemic distribution of plutonium was determined for "healthy" workers who chronically inhaled plutonium at the radiochemical plants of the Mayak Production Association. The data were obtained by radiochemical analysis of soft tissues and bones samples collected upon autopsy of 120 workers who died from acute coronary diseases and accidents. The soft tissue samples were wet-ashed using nitric acid and hydrogen peroxide. Bone samples were ashed in a muffle furnace at 500 degrees C. Plutonium was extracted on anionite and coprecipitated with bismuth phosphate. The precipitation was blended with ZnS powder, and the alpha-activity was measured by ZnS solid scintillation counting in a low-background alpha radiometer. Twenty-five years after the beginning of inhalation exposures, the average percentage of plutonium in the skeleton and liver was 50% and 42% of systemic burden, respectively. A multivariate regression was used to quantify the effects of exposure time, "transportability" of the various compounds, plutonium body content, and age on systemic plutonium distribution. The early retention of plutonium in the liver is assumed to be greater than that in the skeleton. The initial distribution of plutonium between the liver and the skeleton, immediately after entering the circulatory system, was 50:38%, respectively. With time, the fraction of plutonium found in the liver decreased, while the fraction in the skeleton increased at a rate of 0.5% y(-1) of systemic deposition. Exposure time had a greater effect on the relative retention of plutonium in the main organs when compared to age. The statistical estimates that characterized the relative plutonium distribution were less stable for the liver than for the skeleton, likely due to the slower turnover of skeletal tissues and the retention of plutonium in bone. PMID- 11906133 TI - The historical and current application of the FIB-1 model to assess organ dose from plutonium intakes in Mayak workers. AB - One of the objectives of the Joint Coordinating Committee for Radiation Effects Research Project 2.4 is to document the methodology used to determine the radiation doses in workers from the Mayak Production Association who were exposed to plutonium. The doses have been employed in numerous dose response studies measuring both stochastic and deterministic effects. This article documents both the historical (pre-1999) and current ("Doses 1999") methods used by the FIB-1 scientists to determine the doses. Both methods are based on a three-chamber lung model developed by the FIB-1 scientists. This method was developed in partial isolation from the West and has unique characteristics from the more familiar ICRP biokinetic models. Some of these characteristics are the use of empirically based transportability classifications and the parameter modification for chelation-therapy-enhanced excretion data. An example dose calculation is provided and compared to the dose that would be obtained if the ICRP models were used. The comparison demonstrates that the models are not interchangeable and produce different results. PMID- 11906134 TI - Cancer incidence and risk estimation among medical x-ray workers in China, 1950 1995. AB - Cancer incidence (1950-1995) among 27,011 medical diagnostic x-ray workers was compared by means of O/E system with that of 25,782 other medical specialists employed between 1950 and 1980 to provide evidence of human malignant tumors produced by protracted and fractionated exposure to ionizing radiation and to assess resultant cancer risk. Significant cancer risk was seen among diagnostic x ray workers (RR = 1.2, 95% CI: 1.1-1.3). Significantly elevated risks were found for leukemia and cancers of skin, female breast, lung, liver, bladder, and esophagus; the RRs were 2.2, 4.1, 1.3, 1.2, 1.2, 1.8, and 2.7, respectively. The patterns of risk associated with years since beginning x-ray work and with age and calendar year of initial employment suggest that the excesses of leukemia, skin cancer, and female breast cancer-and possibly thyroid cancer-were related to occupational exposure to x rays. Because of a lack of individual dosimetry for Chinese medical x-ray workers (CMXW) before 1985, the dose was reconstructed by physical and biological retrospective dosimetry methods. The cancer risk of CMXW was estimated based on the reconstructed dose. The average cumulative dose for the earlier cohort (employed before 1970) was 551 mGy, and for the later cohort (employed from 1970 to 1980) it was 82 mGy. The RRs of leukemia and solid cancer were significantly high for the earlier cohort: 2.4 for leukemia, 1.2 for solid cancer. But no significant increase of RR was evident for the later cohort. The RR of leukemia was 1.7 and 1.1 for solid cancer. This means a significant cancer risk can be induced by long term fractionated exposure to ionizing radiation when the cumulative dose reaches a certain level. PMID- 11906135 TI - Depleted uranium in Kosovo: an assessment of potential exposure for aid workers. AB - BACKGROUND: During the Kosovo conflict approximately 11 tons of depleted uranium munitions were used against armored targets, predominantly in the west. Potential exposure to uranium amongst employees of the International Red Cross and Red Crescent Movement in western Kosovo was assessed. METHODS: Individuals (n = 31) who had resided at least 3 mo in western Kosovo provided 24-h urine collections and completed an administered questionnaire. Specimens were analyzed for creatinine concentration, and uranium concentration was determined using inductively coupled mass spectrometry. FINDINGS: Subjects ranged in age from 22 to 45 y, and 77% were male. Mean duration of residency was 11 mo, and 14 individuals were in western Kosovo throughout the hostilities. Almost three quarters of subjects reported seeing destroyed tanks or vehicles, predominantly while passing by within a vehicle. Two individuals spent time within 50 m of a destroyed tank or vehicle while outside of a vehicle. Urinary uranium concentrations ranged from 3.5 to 26.9 ng of uranium per liter of urine (median 8.9 ng L(-)). Creatinine normalized values ranged from 2.9 to 21.1 ng of uranium per gram of creatinine (median 7.4 ng g(-1) creatinine). These results fall toward the lower end of urinary uranium determinations made amongst non-exposed populations drawn from a literature review. INTERPRETATION: These results do not indicate an increased exposure to uranium amongst adults living and working in western Kosovo who do not spend time in proximity to destroyed vehicles. Environmental sampling and replication of these results amongst a sample including children and individuals reporting intensive exposure to destroyed vehicles would further develop the exposure assessment. PMID- 11906136 TI - Electromagnetic field standards in Central and Eastern European countries: current state and stipulations for international harmonization. AB - Electromagnetic field standards in the West are based on well-established acute biological effects that could be considered as signaling a potentially adverse health effect. The specific absorption rate, which is proportional to the tissue heating (thermal effects), represents the basic restriction of exposure to Radio Frequency (RF) fields. On the other hand, Eastern European standards are designed to protect from potential non-thermal effects that might be caused by chronic exposure to very low intensities, where a so-called "power load" (a product of field intensity and duration of exposure) represents the basic limitation. Thus, electromagnetic field standards in Eastern European countries differ considerably from those which are proposed by the International Commission of Non-ionizing Radiation Protection and the Standards Coordinating Committee 28 of the Institute of Electrical and Electronics Engineers, Inc. In the present paper, the strategies for development of exposure limit values in electromagnetic fields standards currently in force in Eastern and Central European countries are discussed. Some differences as well as similarities of the national health and safety standards and the main obstacles to harmonization of these standards with those being established by Western national and international organizations and agencies are presented. PMID- 11906137 TI - Assessment of guidelines for limiting exposures to emf using methods of probabilistic risk analysis. AB - Allowable limits of human exposure to radiofrequency fields commonly include a "factor of safety," typically between 10 to 50, which is somewhat arbitrary. The broad objective in our work is to assess radiofrequency exposure limits, hazard thresholds, and safety factors using methods of probabilistic risk analysis. We focus our analysis on the variables affecting peak radiofrequency specific energy absorption rate (SAR) values in the brain from digital mobile telephones operating at approximately 900 MHz. As SAR is defined as a product of positive random variables, it is not unreasonable to assume that SAR has a lognormal distribution. Our analysis of component SAR variables such as conductivity and permittivity of grey brain matter and radiated field strengths using experimental and numerical modeling data strongly supports our hypothesis that SAR values are distributed lognormally. It then follows that the probability that the SAR exceeds a certain threshold can be derived directly and is shown to be very low for handset SARs relative to presently allowable standard limits. PMID- 11906138 TI - Gamma and neutrino radiation dose from gamma ray bursts and nearby supernovae. AB - Supernovae and gamma ray bursts are exceptionally powerful cosmic events that occur randomly in space and time in our galaxy. Their potential to produce very high radiation levels has been discussed, along with speculation that they may have caused mass extinctions noted from the fossil record. It is far more likely that they have produced radiation levels that, while not lethal, are genetically significant, and these events may have influenced the course of evolution and the manner in which organisms respond to radiation insult. Finally, intense gamma radiation exposure from these events may influence the ability of living organisms to travel through space. Calculations presented in this paper suggest that supernovae and gamma ray bursts are likely to produce sea-level radiation exposures of about I Gy with a mean interval of about five million years and sea level radiation exposures of about 0.2 Gy every million years. Comets and meteors traveling through space would receive doses in excess of 10 Gy at a depth of 0.02 m at mean intervals of 4 and 156 million years, respectively. This may place some constraints on the ability of life to travel through space either between planets or between planetary systems. Calculations of radiation dose from neutrino radiation are presented and indicate that this is not a significant source of radiation exposure for even extremely close events for the expected neutrino spectrum from these events. PMID- 11906139 TI - The studies on radiological limits of color-glazed tiles used in home decoration. AB - This study was carried out to lay down the radiological limits of color-glazed tiles used in home decoration. The activity concentrations of various end products and raw materials as well as processed materials were measured using gamma spectroscopy. 222Rn exhalation rates from the surface of color-glazed tiles were measured using charcoal canister method. Levels of exposure to alpha ray and beta ray from the surface of glazed tiles were measured by surface alpha and beta contaminant instrument. The results show a great difference between the radioactive levels of the surface glaze and the matrix of color-glazed tiles. The 222Rn exhalation rates from the surface of color-glazed tiles are in the range of 10(-3) - 10(-4) Bq m(-2) s(-1). The concentrations of some natural radionuclides in glaze exceed the exempt limits of International Basic Safety Standards for Protection against Ionizing Radiation Sources (IAEA 1997). The limits for color glazed tiles were deduced according to these data and the theory of UNSCEAR (1993). The activity concentration of 226Ra (Bq kg(-1)) in the glaze of color glazed tiles should be in the range of A(226Ra) < or = 1,000 (Bq kg(-1)), and, at the same time, the specific activity of the natural radionuclides (Bq kg(-1)) should be in the range of A(232Th)/230 + A(226Ra)/310 + A(40K)/3,500 < or = 1. PMID- 11906140 TI - Radiation hormesis: challenging LNT theory via ecological and evolutionary considerations. AB - Ecological and evolutionary considerations suggest that radiation hormesis is made up of two underlying components. The first (a) is background radiation hormesis based upon the background exposure to which all organisms are subjected throughout evolutionary time. The second and much larger component (b) is stress derived radiation hormesis arising as a protective mechanism derived from metabolic adaptation to environmental stresses throughout evolutionary time especially from climate-based extremes. Since (b) > > (a), hormesis for ionizing radiation becomes an evolutionary expectation at exposures substantially exceeding background. This biological model renders linear no-threshold theory invalid. Accumulating evidence from experimental organisms ranging from protozoa to rodents, and from demographic studies on humans, is consistent with this interpretation. Although hormesis is not universally accepted, the model presented can be subjected to hypothesis-based empirical investigations in a range of organisms. At this stage, however, two consequences follow from this evolutionary model: (1) hormesis does not connote a value judgement usually expressed as a benefit; and (2) there is an emerging and increasingly convincing case for reviewing and relaxing some recommended radiation protection exposure levels in the low range. PMID- 11906141 TI - A case of repeated accidental inhalation contamination of a male subject with 137Cs. AB - Two consecutive incidents of internal contamination occurred within a 13-mo period when a male worker (56 y, 70 kg) briefly entered and stayed (during a breakdown of control apparatus for air contamination) in a radioisotope storeroom with air contaminated by an explosion of old ampules containing 137Cs solution. Monitoring of the first contamination began on day 34 and that of the second one within 1 h after inhalation. According to the two-exponential model, long-term biological half-times of 92 and 93 d were obtained for the first and second contaminations, respectively. The short-term biological half-time for the second contamination was calculated as 3.0 d. The mean value of the fraction of 137Cs excreted in daily urine to that in total excreta was 0.88 for the first contamination and 0.89 for the second one. PMID- 11906142 TI - Environmental gamma-ray dose rate in Aomori Prefecture, Japan. AB - Japan's first commercial nuclear fuel cycling facilities, including a spent nuclear fuel reprocessing plant, are now under construction in Rokkasho, Aomori Prefecture (prefecture--an area of administration similar to a county in the U.S.). The reprocessing plant is due to be completed by 2004. We surveyed indoor and outdoor environmental gamma-ray dose rates throughout Aomori Prefecture from 1992 to 1996 to get background data before operation of the plant. Glass dosimeters were used to measure cumulative gamma-ray dose rate. The outdoor gamma ray dose rates were measured at 109 locations in the prefecture. The indoor gamma ray dose rates were measured at 81 locations, which were generally in a dwelling near the location of an outdoor measurement. The contribution of radionuclides in the ground to the outdoor dose rate was estimated by using in-situ gamma-ray spectrometry with a germanium detector. The spectra were measured at 20 locations used for the glass dosimeter measurements. The outdoor gamma-ray dose rate was higher in the Tsugaru area (western part of the prefecture) than in the Nanbu area (eastern part). Means of the dose rate were 28, 31, and 25 nGy h(-1) for the whole prefecture and Tsugaru and Nanbu areas, respectively. The dose rates in winter were lower than those in the other seasons due to the shielding effect of snow on the ground. Mean contributions of uranium series, thorium series and 40K to the dose rates were 7.7, 8.2, and 9.3 nGy h(-1), respectively. The indoor dose rate was generally higher than the outdoor one, and the mean ratio of indoor to outdoor dose rates was 1.42. Means of indoor gamma-ray dose rate were 41, 37, and 43 nGy h(-1) for the whole prefecture and Tsugaru and Nanbu areas, respectively. The average effective dose rate to people in the prefecture was estimated to be 0.24 mSv y(-1). PMID- 11906143 TI - Development of a routine 125I bioassay program for athyrotic individuals using a pseudo uptake retention function. AB - Individuals working in iodine production require bioassay to determine if intakes have occurred. This is both to determine dose received for regulatory purposes and to verify whether workplace controls limiting the spread of contamination are adequate. Thyroid monitoring is commonly used as a bioassay technique to detect isotopes of iodine. If an individual performing iodine processing does not have a thyroid gland, other means must be used to determine intake and infer dose. Data was obtained from a previously published thesis that attempted to verify a model for absorption and retention of iodine by athyrotic individuals. These data were reevaluated to determine a pseudo uptake retention function. This analysis does not attempt to identify a biokinetic model, only to describe excretion of iodine and calculate an intake. Once the pseudo uptake retention function was derived, it was combined with the standard respiratory and gastrointestinal tract models as an inhalation intake retention function. A periodic urine bioassay protocol has been designed using the intake retention function described above and a conservative dose coefficient derived using organ dose coefficients for reference man, excluding the thyroid, and the appropriate weighting factors. PMID- 11906144 TI - General approach to protection against non-ionizing radiation. PMID- 11906145 TI - Advances in the voltammetric analysis of small biologically relevant compounds. AB - The problems associated with attempting to apply voltammetric techniques to the analysis of biologically relevant organics within complex media are identified and, through reviewing the very recent literature (1999-mid-2001), possible solutions are described. The boundaries of the search were limited to research targeted at the resolution of specific problems, associated with quantitative determinations. Various strategies have emerged to counter problems of poor sensitivity and selectivity and these have been summarized and critically appraised. Where possible, the characteristics of each approach have been distilled into a table format to ease comparison. Emphasis has been placed on the collation of information that will improve the intrinsic electrode response and as such should be of value to those interested in pursuing electroanalytical methodologies regardless of context. PMID- 11906147 TI - Accessing single nucleotide polymorphisms in genomic DNA by direct multiplex polymerase chain reaction amplification on oligonucleotide microarrays. AB - This study introduces a DNA microarray-based genotyping system for accessing single nucleotide polymorphisms (SNPs) directly from a genomic DNA sample. The described one-step approach combines multiplex amplification and allele-specific solid-phase PCR into an on-chip reaction platform. The multiplex amplification of genomic DNA and the genotyping reaction are both performed directly on the microarray in a single reaction. Oligonucleotides that interrogate single nucleotide positions within multiple genomic regions of interest are covalently tethered to a glass chip, allowing quick analysis of reaction products by fluorescence scanning. Due to a fourfold SNP detection approach employing simultaneous probing of sense and antisense strand information, genotypes can be automatically assigned and validated using a simple computer algorithm. We used the described procedure for parallel genotyping of 10 different polymorphisms in a single reaction and successfully analyzed more than 100 human DNA samples. More than 99% of genotype data were in agreement with data obtained in control experiments with allele-specific oligonucleotide hybridization and capillary sequencing. Our results suggest that this approach might constitute a powerful tool for the analysis of genetic variation. PMID- 11906146 TI - An ultraviolet spectrophotometric assay for measuring lipase activity using long chain triacyglycerols from Aleurites fordii seeds. AB - In this study, we designed a specific, continuous, and sensitive UV spectrophotometric lipase assay using natural triacylglycerols (TAGs) from the Aleurites fordii seed oil (tung oil). alpha-Eleostearic acid (9,11,13-cis, trans,trans-octadecatrienoic acid) is the main fatty acid component (it accounts for up to 70%) of the TAGs from tung oil. The conjugated triene present in alpha eleostearic acid constitutes an intrinsic chromophore, which confers strong UV absorption properties on both the free fatty acid and the TAGs from tung oil. The lipase assay is based on the difference between the apparent molar extinction coefficients of the two types of alpha-eleostearic acid present, that which is esterified into TAGs and that which is released into the reaction medium. This difference is responsible for the variations in the UV absorption spectrum of the reaction medium occurring upon enzymatic TAGs hydrolysis. Using the purified lipase from Thermomyces lanuginosa (TLL) and the detergent sodium taurodeoxycholate (NaTDC, 4 mM), it was established that the most suitable method of measuring lipolysis consisted of monitoring the decrease in the OD at 292 nm, which was linear with time and proportional to the amount of lipase added. In order to be able to estimate the specific activity of TLL, we determined an apparent molar extinction coefficient of alpha-eleostearic acid (epsilon = 13,900 M(-1) cm(-1)) under the assay conditions. Amounts of pure TLL as small as 1 ng can be easily detected in the presence of 4 mM NaTDC. Interestingly, the NaTDC concentration can be decreased as far as 0.05 mM. In comparison with other well known methods of lipase assay, the detection limit of this new method is 100-fold lower than with the pH-stat method and similar to that of a fluorescent assay recently developed at our laboratory. PMID- 11906148 TI - A method for functional mapping of protein-protein binding domain by preferential amplification of the shortest amplicon using PCR. AB - We have developed a novel method for rapid and empirical mapping of the protein interaction domain using a unique and atypical PCR-based amplification and a conventional yeast two-hybrid system. The modified PCR, designated as PASA-PCR, enables preferential amplification of the shortest amplicon from a complex expression library. PASA-PCR consists of reiterative cycles of denaturation of template DNAs and extremely abbreviated annealing/extension of primers to prevent their complete extension in a single cycle, followed by conventional amplification cycles. In PASA-PCR, the shortest (ranging from 400 to 1000 bp) amplicon is amplified almost exclusively from templates of various amplicon sizes. In addition, the frequency of in vitro recombination can be increased using low cooling rates (<0.5 degrees C/s) between the denaturation and annealing/extension steps, which was helpful in generating precisely trimmed protein-coding regions. Identification of Spc19-binding region of Spc34, which is a component of yeast's spindle pole body, was achieved by a combination of the yeast two-hybrid system and PASA-PCR. PMID- 11906149 TI - Enzyme-based sensor arrays for rapid characterization of complex disaccharide solutions. AB - An enzyme-based sensor array has been developed to detect multiple disaccharides in aqueous solutions. Porous agarose beads, derivatized with enzymes for assaying disaccharides, are localized within wells etched into a silicon chip in a regular 5 x 7 array. Each well is individually addressable and acts as a microanalysis chamber where sample solution passes through the agarose matrix and is exposed to the enzymes. Detection is achieved by observing the increase in absorbance of a quinoneimine dye produced during the reaction. This technique is used to quantify the disaccharides lactose, sucrose, and maltose and the monosaccharide glucose. Preexisting glucose in the sample complicates multicomponent sensing but can be accounted for by including a glucose sensor in the array. This detection strategy is applied to the simultaneous analysis of these sugars in several beverages. PMID- 11906150 TI - Continuous spectrometric assays for glutaminyl cyclase activity. AB - The enzymatic conversion of one chromogenic substrate, l-glutamine-p nitroanilide, and two fluorogenic substrates, l-glutaminyl-2-naphthylamide and l glutaminyl-4-methylcoumarinylamide, into their respective pyroglutamic acid derivatives by glutaminyl cyclase (QC) was estimated by introducing a new coupled assay using pyroglutamyl aminopeptidase as the auxiliary enzyme. For the purified papaya QC, the kinetic parameters were found to be in the range of those previously reported for other glutaminyl peptides, such as Gln-Gln, Gln-Ala, or Gln-tert-butyl ester. The assay can be performed in the presence of ammonia up to a concentration of 50 mM. Increasing ionic strength, e.g., potassium chloride up to 300 mM, resulted in an increase in enzymatic activity of about 20%. This is the first report of a fast, continuous, and reliable determination of QC activity, even in the presence of ammonium ions, during the course of protein purification and enzymatic analysis. PMID- 11906151 TI - Theoretical and experimental dissection of competitive PCR for accurate quantification of DNA. AB - We frequently use competitive PCR in the plateau phase in quantifying DNA species with a small number of cells. However, the basic issues of this method are poorly understood. Here, first we analyze this method theoretically under a generalized condition that competitor and target DNA products accumulate with different amplification efficiencies. We show a theoretical reason that competitive PCR might quantify DNA more accurately during the plateau phase than during the exponential phase. Second, we demonstrate that the theoretical predictions are supported by the experimental results of beta-globin gene amplification using the lysates of human diploid fibroblast WS1 cells. We also demonstrate that we can correctly quantify target DNA by keeping the starting concentration of target DNA close to a constant preset value while using a constant number of PCR cycles and by using WS1 cells as control. Finally, we show the experimental errors in routine measurements of c-myc copy number/cell in human leukemia HL-60 cells with various levels of c-myc multiplication. The number of c-myc copies/cell was determined with an error rate of less than 10%, where agarose gel bands were stained with ethidium bromide for the product quantitation. PMID- 11906152 TI - Mechanistic study of the oxidation of caffeic acid by digital simulation of cyclic voltammograms. AB - The oxidation mechanism of caffeic acid (CAF) has been studied by means of cyclic voltammetry with the plastic formed carbon or glassy carbon electrode. CAF gives a well-developed two-electron reversible wave in acidic media, whereas it shows an irreversible behavior, i.e., a decrease of the rereduction peak, in less acidic media, suggesting that the oxidation of CAF follows an irreversible chemical reaction(s). Digital simulation analyses based on different oxidation mechanisms have been performed for the voltammograms obtained with the GC electrode in 1:1 (v/v) water:ethanol solutions. The results clearly show that the seeming two-electron oxidation of CAF occurs stepwise via one-electron processes, each of which follows an irreversible chemical reaction. It has also been suggested that the semiquinone radical as an intermediate of the one-electron oxidation should play an important role in the oxidation reaction. Evaluations of the rate constants for the chemical reactions have further suggested that the chemical reactions are dimerization reactions. PMID- 11906153 TI - Ultracentrifugation micromethod for preparation of small experimental animal lipoproteins. AB - Sequential flotation ultracentrifugation is commonly used in the preparation of plasma lipoproteins. However, protocols often require prolonged centrifugation time (48-72 h) and large plasma volumes (2-20 ml), which makes them unsuitable for studies on small laboratory animals. Although analytical techniques such as FPLC have often small sample requirements, further fraction analysis is often limited to the small fraction volume obtained. A sequential ultracentrifugation micromethod is described to obtain rat lipoprotein fractions from 400 microl of plasma in a cumulative centrifugation time of 7.5 h. Fraction volumes were determined and densities were adjusted to those of rat plasma lipoproteins. Polyacrylamide gel electrophoresis and enzymatic measurements of triglycerides, total cholesterol, and phospholipids were used to assess the purity of the lipoprotein fractions. The results were compared with those obtained from a classical sequential ultracentrifugation protocol. The micromethod presented here can be further adapted to other experimental animal species with little modifications. PMID- 11906154 TI - A mathematical analysis using fractals for binding interactions of nuclear estrogen receptors occurring on biosensor surfaces. AB - A mathematical approach using fractal concepts is presented for modeling the binding and dissociation interactions between analytes and nuclear estrogen receptors (ER) occurring on surface plasmon resonance biosensor chip surfaces. A kinetic knowledge of the binding interactions mediated by ER would help in better understanding the carcinogenicity of these steroidogenic compounds and assist in modulating these reactions. The fractal approach is applied to analyte-ER interaction data obtained from literature. Numerical values obtained for the binding and dissociation rate coefficients are linked to the degree of roughness or heterogeneity (fractal dimension, D(f)) present on the biosensor surface. For example, a single-fractal analysis is used to describe the binding and dissociation phases for the binding of estradiol and ERalpha in solution to clone 31 protein immobilized on a biosensor chip (C-S. Suen et al., 1998, J. Biol. Chem. 273(42), 27645-27653). The binding and the dissociation rate coefficients are 27.57 and 8.813, respectively, and the corresponding fractal dimensions are 1.986 and 2.268, respectively. In some examples dual-fractal models were employed to obtain a better fit of either the association or the dissociation phases or for both. Predictive relationships are developed for (a) the binding and the dissociation rate coefficients as a function of their respective fractal dimensions and (b) the ratio K(A) (= k/k(d)) as a function of the ratio of the fractal dimensions (D(f)/D(fd)). The analysis should provide further physical insights into the ER-mediated interactions occurring on biosensor and other surfaces. PMID- 11906155 TI - Use of a meltable polyacrylamide matrix for sodium dodecyl sulfate-polyacrylamide gel electrophoresis in a procedure for N-glycan analysis on picomole amounts of glycoproteins. PMID- 11906156 TI - Elimination of primer-dimer artifacts and genomic coamplification using a two step SYBR green I real-time RT-PCR. PMID- 11906158 TI - One-step unidirectional cloning of tandem repeats of DNA fragments: an application for fusion protein production. PMID- 11906157 TI - Effects of sialic acid substitutions on recognition by Sambucus nigra agglutinin and Maackia amurensis hemagglutinin. PMID- 11906159 TI - Comet assay coupled to repair enzymes for the detection of oxidative damage to DNA induced by low doses of gamma-radiation: use of YOYO-1, low-background slides, and optimized electrophoresis conditions. PMID- 11906160 TI - The involvement of molybdenum in life. AB - Quite extraordinarily molybdenum is an essential element in life for the uptake of nitrogen from both nitrogen gas and nitrate, yet it is a relatively rare heavy trace element. It also functions in a few extremely important oxygen-atom transfer reactions at low redox potential. This review poses the question "Why does life depend upon molybdenum?" The answer has to be based upon the availability of the element and on chemical superiority in carrying out the essential tasks. We illustrate here the peculiarities of molybdenum chemistry and how they have become part of certain enzymes. The uptake and incorporation of molybdenum are dependent on its availability, selective pumps, and carriers (chaperones), but 4.5 x 10(9) years ago molybdenum was not available when both tungsten and vanadium or even iron were possibly used in its place. While these possibilities are explored, they leave many unanswered questions concerning the selection today of molybdenum. PMID- 11906161 TI - DOC-2/DAB2 is the binding partner of myosin VI. AB - Myosin VI is a molecular motor that moves processively along actin filaments and is believed to play a role in cargo movement in cells. Here we found that DOC 2/DAB2, a signaling molecule inhibiting the Ras cascade, binds to myosin VI at the globular tail domain. DOC-2/DAB2 binds stoichiometrically to myosin VI with one molecule per one myosin VI heavy chain. The C-terminal 122 amino acid residues of DOC-2/DAB2, containing the Grb2 binding site, is identified to be critical for the binding to myosin VI. Actin gliding assay revealed that the binding of DOC-2/DAB2 to myosin VI can support the actin filament gliding by myosin VI, suggesting that it can function as a myosin VI anchoring molecule. The C-terminal domain but not the N-terminal domain of DOC-2/DAB2 functions as a myosin VI anchoring site. The present findings suggest that myosin VI plays a role in transporting DOC-2/DAB2, a Ras cascade signaling molecule, thus involved in Ras signaling pathways. PMID- 11906162 TI - Glucose transport by osmotic shock and vanadate is impaired by glucosamine. AB - In 3T3-L1 adipocytes, we previously reported that glucosamine impairs insulin stimulation of glucose transport, which is accompanied by impaired insulin stimulation of serine/threonine kinase Akt. To examine the role of Akt in glucosamine-induced insulin resistance, we investigated time course for insulin stimulation of Akt activity and glucose transport during recovery from glucosamine-induced insulin resistance. After induction of insulin resistance by glucosamine, we washed cells to remove glucosamine and incubated them for various times. After one hour, insulin stimulated-glucose transport was significantly increased and continued to increase up to 6-24 h. Insulin stimulation of Akt, however, did not increase after 1-3 h and began to slightly increase after 6 h. Next, we investigated effects of osmotic shock and vanadate on glucose transport in glucosamine-treated cells and found that glucosamine completely inhibited their actions in these cells. These data suggest that an Akt-independent mechanism is operative in glucosamine-induced insulin resistance and glucosamine impairs glucose transport stimulated by various stimuli involving and not involving Akt activation. PMID- 11906163 TI - Phenobarbitone-mediated translocation of the cytosolic proteins interacting with the 5'-proximal region of rat liver CYP2B1/B2 gene into the nucleus. AB - The positive element (PE) (-69 to -98 bp) within the 5'-proximal region of the CYP2B1/B2 gene (+1 to -179 bp) of rat liver is essential for phenobarbitone (PB) response and gives a single major complex with the rat liver cytosol in gel shift analysis. This complex corresponds to complex I (top) of the three complexes given by the nuclear extracts. PB treatment of rats leads to a decrease in complex I formation with the cytosol and PE and an increase in the same with the nuclear extract in gel shift analysis. Both the changes are counteracted by simultaneous okadaic acid administration. The nuclear protein giving rise to complex I has been isolated and has an M(r) of 26 kDa. The cytosolic counterpart consists of two species, 26 and 28 kDa, as revealed by Southwestern blot analysis using labeled PE. It is concluded that PB treatment leads to the translocation accompanied by processing of the cytosolic protein species into the nucleus that requires protein dephosphorylation. It is suggested that PB may exert a global regulation on the transcription of many genes by modulating the phosphorylation status of different protein factors involved in transcriptional regulation. PMID- 11906164 TI - Motor discoordination in mutant mice lacking junctophilin type 3. AB - Junctional complexes between the plasma membrane and endoplasmic reticulum (ER), often called "subsurface cisternae" or "peripheral coupling," are shared by excitable cells. These junctional membranes probably provide structural foundation for functional crosstalk between cell-surface and intracellular ionic channels. Our current studies have indicated that junctophilins (JPs) take part in the formation of junctional membrane complexes by spanning the ER membrane and interacting with the plasma membrane. Of the JP subtypes defined, JP type 3 (JP 3) is specifically expressed in neurons in the brain. It has been currently reported that triplet repeat expansions in the JP-3 gene are associated with Huntington's disease-like symptoms including motor disorder in human. To survey the physiological role of JP-3, we generated the knockout mice. The JP-3-knockout mice grew and reproduced normally, and we did not observe any morphological abnormality in the mutant brain. In the behavioral study, the mutant mice showed impaired performance specifically in balance/motor coordination tasks. Although obvious defects could not be observed in excitatory transmission among cerebellar neurons from the mutant mice, the data indicate that JP-3 plays an active role in certain neurons involved in motor coordination. PMID- 11906165 TI - Phospholipase D regulates calcium oscillation frequency and nuclear factor-kappaB activity in histamine- stimulated human endothelial cells. AB - Histamine stimulates [Ca(2+)](i) oscillations in human aortic endothelial cells (HAEC), the frequency of which regulates the activity of nuclear factor-kappaB (NF-kappaB). This study was performed to determine whether phospholipase D (PLD) is involved in this signaling pathway. At a concentration of 1 microM, which stimulates [Ca(2+)](i) oscillations in this cell type, histamine initiated a twofold increase in [(32)P]phosphatidybutanol (PBt), an index of PLD activity as early as 5 min after stimulation. During established [Ca(2+)](i) oscillations induced by 1 microM histamine, 0.3% n-butanol, which "functionally" redirects phosphatidic acid formed by PLD to PBt, decreased [Ca(2+)](i) oscillation frequency by approximately 50% and produced a similar reduction in NF-kappaB activity. In the presence of the inositol 1,4,5-trisphosphate receptor blocker xestospongin C, which itself decreases the frequency of histamine-stimulated [Ca(2+)](i) oscillations, n-butanol produced a further decrease in oscillation frequency that was not associated with an additional reduction in NF-kappaB activity. This study shows that activation of PLD by histamine regulates [Ca(2+)](i) oscillation frequency and NF-kappaB activity in HAEC. PMID- 11906166 TI - A role for the common GTP-binding protein in coupling of chromosome replication to cell growth and cell division. AB - Homologues of CgtA, the common GTP-binding protein of Vibrio harveyi, are present in diverse organisms ranging from bacteria to humans. In bacteria, proteins homologous to CgtA form a subfamily of small GTP-binding proteins, called Obg/Gtp1. Similarity between bacterial members of this subfamily and their eukaryotic homologues is as high as about 50%. Nevertheless, specific functions of these proteins remain largely unknown. Genes coding for CgtA-like proteins are essential in almost all species of bacteria. The only known exception is V. harveyi, whose cells survive disruption of the cgtA gene. Therefore, the V. harveyi cgtA insertional mutant is a very useful tool for studies on functions of CgtA. Here we demonstrate that under normal growth conditions, cells of the cgtA mutant are slightly larger than wild-type cells, whereas indirect inhibition of DNA replication initiation by addition of rifampicin results in significantly higher differences in average cell size between these two strains as measured by flow cytometry. These differences decreased when cell division was inhibited by cephalexin. DNA synthesis per cell mass was found to be increased in the cgtA mutant relative to wild-type V. harveyi strain, whereas the mutant cells grew slower than bacteria with functional cgtA gene. Kinetics of DNA replication after inhibition of cell division was also considerably different in wild-type and cgtA mutant strains. These results suggest that the cgtA gene product plays a role in coupling of DNA replication to cell growth and cell division. PMID- 11906167 TI - Molecular basis of the voltage-dependent gating of TREK-1, a mechano-sensitive K(+) channel. AB - TREK-1 is a member of the mammalian two P domain K(+) channel family. Mouse TREK 1 activity, in transiently transfected COS cells, is reduced at negative resting membrane potentials by both an external Mg(2+) block and an intrinsic voltage dependent gating mechanism leading to a strong outward rectification. Deletional and chimeric analysis demonstrates that the carboxy terminal domain of TREK-1, but not the PKA phosphorylation site S333, is responsible for voltage-dependent gating. Since the same region is also critically required for TREK-1 mechano gating, both mechanisms might be functionally linked. Preferential opening of TREK-1 at depolarized potentials will greatly affect action potential duration, recovery from inactivation and neuronal repetitive firing activity. PMID- 11906169 TI - Exposure to power frequency magnetic fields suppresses X-ray-induced apoptosis transiently in Ku80-deficient xrs5 cells. AB - In an attempt to determine whether exposure to extremely low frequency (ELF) electromagnetic fields can affect cells, Ku80-deficient cells (xrs5) and Ku80 proficient cells (CHO-K1) were exposed to ELF electromagnetic fields. Cell survival, and the levels of the apoptosis-related genes p21, p53, phospho-p53 (Ser(15)), caspase-3 and the anti-apoptosis gene bcl-2 were determined in xrs5 and CHO-K1 cells following exposure to ELF electromagnetic fields and X-rays. It was found that exposure of xrs5 and CHO-K1 cells to 60 Hz ELF electromagnetic fields had no effect on cell survival, cell cycle distribution and protein expression. Exposure of xrs5 cells to 60 Hz ELF electromagnetic fields for 5 h after irradiation significantly inhibited G(1) cell cycle arrest induced by X rays (1 Gy) and resulted in elevated bcl-2 expression. A significant decrease in the induction of p53, phospho-p53, caspase-3 and p21 proteins was observed in xrs5 cells when irradiation by X-rays (8 Gy) was followed by exposure to 5 mT ELF magnetic fields. Exposure of xrs5 cells to the ELF electromagnetic fields for 10 h following irradiation significantly decreased X-ray-induced apoptosis from about 1.7% to 0.7%. However, this effect was not found in CHO-K1 cells within 24 h of irradiation by X-rays alone and by X-rays combined with ELF electromagnetic fields. Exposure of xrs5 cells to 60 Hz ELF electromagnetic fields following irradiation can affect cell cycle distribution and transiently suppress apoptosis by decreasing the levels of caspase-3, p21, p53 and phospho-p53 and by increasing bcl-2 expression. PMID- 11906168 TI - Caspase-8 gene transduction augments radiation-induced apoptosis in DLD-1 cells. AB - Caspase-8 is a member of the cysteine protease family that plays a critical role in death receptor-mediated apoptosis. We previously demonstrated that adenovirally transduced caspase-8 efficiently induced apoptosis in tumor cells (Shinoura et al. (2000) Hum. Gene Ther. 11, 1123-1137). However, to ensure safety in clinical applications some devise for minimization of the dose of adenoviral vector required for sufficient antitumor effect is needed. In this study, we evaluated the proapoptotic effect in DLD-1 colon cancer cells of a combination of low-dose infection with an adenoviral vector expressing caspase-8 and X-ray irradiation. Under these conditions, X-ray irradiation strongly induced apoptosis whereas irradiation without transduction only had a trace proapoptotic effect. Overexpression of bcl-xL strongly blocked the activation of caspase-8 and induction of apoptosis, suggesting that adenovirally transduced caspase-8 was activated at a point downstream of mitochondria. This combination strategy may be a useful modality for gene therapy of colorectal cancer. PMID- 11906170 TI - Role of the conserved DRY motif on G protein activation of rat angiotensin II receptor type 1A. AB - To delineate the functional importance of the highly conserved triplet amino acid sequence, Asp-Arg-Tyr (DRY) among G protein-coupled receptors in the second intracellular loop, these residues of rat angiotensin II (Ang II) receptor type 1A (AT(1A)) were changed by alanine or glycine by site-directed mutagenesis. These mutant receptors were stably expressed in CHO-K1 cells, and the binding of Ang II, GTP effect, InsP(3) production, and the acidification of the medium in response to Ang II were determined. The effects of GTPgammaS on Ang II binding in the mutant receptors D125A and D125G were markedly reduced. InsP(3) production of the mutant D125A, D125G, R126A, and R126G was markedly reduced. Extracellular acidification of D125A was not distinguishable from untransfected CHO-K1 cells. Mutant Y127A was able to produce InsP(3) and acidify medium comparable with wild type AT(1A). These results indicate as follows; Asp(125) is essential for intracellular signal transduction involving G protein coupling, Arg(126) is essential for coupling of G(q) protein but not other G proteins, and Tyr(127) is not important for G protein coupling. PMID- 11906171 TI - Identification of 2,3,7,8-tetrachlorodibenzo-p-dioxin-responsive genes in mouse liver by serial analysis of gene expression. AB - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is one of the most toxic environmental pollutants that causes various biological effects on mammals. To identify the genes involved in hepatotoxicity and hepatocarcinogenesis induced by TCDD, we have conducted here serial analysis of gene expression of mouse liver 7 days after treatment with a single oral dose of 20 microg TCDD/kg body weight. We have sequenced total of 113,067 tags, including 56,420 tags and 56,647 tags from normal liver and TCDD-treated liver library, respectively. Statistical analysis showed that TCDD significantly altered 346 transcripts (p < 0.05) including 94 ESTs. The genes regulated by TCDD were not only the genes encoding drug metabolizing enzymes and stress response genes but also a wide variety of genes encoding cytoskeleton related proteins, signal transduction, and plasma proteins. This comprehensive gene expression analysis would provide novel genes that may help to clarify the mechanism of TCDD effects on mammalian liver, and also give a new approach for prevention and treatment. PMID- 11906172 TI - Identification and characterization of soluble isoform of fibroblast growth factor receptor 3 in human SaOS-2 osteosarcoma cells. AB - We have previously reported the alternatively spliced transcripts of fibroblast growth factor 3 (FGFR3 ATs and MTs) derived by aberrant splicing and usage of cryptic splicing sites. Here, we describe a soluble variant of FGFR3 (FGFR3 AT III) arising from skipping exons 8, 9, and 10 in human SaOS-2 osteosarcoma cell. This splicing event leads to the generation of an mRNA encoding a FGFR3 in which the COOH-terminal portion of the Ig-like-III domain and transmembrane domain are deleted while the remainder of the mature molecule is fused in-frame to the COOH terminal cytoplasmic kinases domains. Sf9 cells transfected with the corresponding cDNA express the soluble form of FGFR3 AT-III into the condition medium and its secreted form was able to bind both FGF-1 and FGF-2 leading to loss of ligand binding specificity. These results indicate that the FGFR3 AT-III mRNAs are transcribed due to exon skipping with altered ligand binding specificity. These results suggest that the presence of soluble transcripts of FGFRs gene is a common feature due to mRNA splicing and this splicing plays an important role in the regulation of FGFRs function. PMID- 11906173 TI - Attenuation of transforming growth factor beta-induced growth inhibition in human hepatocellular carcinoma cell lines by cyclin D1 overexpression. AB - Transforming growth factor-beta1 (TGF-beta1) causes growth inhibition in many cell types. Since its role in the outgrowth of human hepatocellular carcinoma (HCC) is not clearly understood, we investigated the growth inhibitory effects of TGF-beta1, the genetic and molecular integrity of TGF-beta receptors, and the expression levels of cell cycle regulating proteins in 11 human HCC cell lines. Of 11 cell lines, 3 (27%) showed growth inhibition to TGF-beta1, whereas the other 8 cell lines did not. We performed Southern and Northern analysis of TGF beta type I and II receptors and examined poly-adenine track mutation of the TGF beta type II receptor, but failed to find any genetic mutation. The transcriptional induction of plasminogen activator inhibitor-1 and p21(WAF1/CIP1) by TGF-beta were detected in all HCC cell lines, implying that the molecular integrity of the TGF-beta receptors might be intact. The amplification and overexpression of cyclin D1 gene was detected in 4 (50%) of 8 HCC cells that showed resistance to TGF-beta1. The suppression of cyclin D1 expression with antisense cyclin D1 facilitated the TGF-beta1-triggered growth inhibition in a TGF-beta1 resistant HCC cell line containing amplified cyclin D1 gene. In conclusion, the overexpression of cyclin D1 may be responsible for the attenuation of TGF-beta1 induced growth inhibition in some HCC cells. PMID- 11906174 TI - Biliary anionic peptide fraction and apoA-I regulate intestinal cholesterol uptake. AB - Evidence is now in favor of protein-facilitated mechanisms for the intestinal cholesterol absorption. Here we report that the unesterified cholesterol uptake by rat jejunal brush border membrane vesicles (BBMVs) is efficient, saturable, and protein-mediated. The human apolipoproteins biliary anionic peptide factor (APF) and A-I (apoA-I) up-regulate micellar cholesterol uptake in a dose dependent manner, but for all tested concentrations (0.1-20 microM), the lipid free APF was more efficient than apoA-I. This uptake stimulation was suppressed after addition of Pabs directed to the external lipid-binding domain of the CLA 1/SR-BI and reduced by Pabs directed to the external loop of CD36. Thus, CLA-1/SR BI and to a lesser extent CD36 are involved in the regulation of intestinal cholesterol uptake. APF, the main protein bound to biliary lipids, is likely one of their physiological effectors. As APF is an unesterified cholesterol carrier, it could facilitate the intestinal absorption of biliary cholesterol. PMID- 11906175 TI - The function of OmpA in Escherichia coli. AB - Outer membrane protein A (OmpA) is a major protein in the Escherichia coli outer membrane. In this study, the function of OmpA in E. coli stress survival was examined. An E. coli K1 ompA-deletion mutant was significantly more sensitive than that of its parent strain to sodium dodecyl sulfate (SDS), cholate, acidic environment, high osmolarity, and pooled human serum. A number of amino acid changes at the extracellular loops of OmpA did not affect the viability of E. coli, while short peptide insertions in the periplasmic turns of the OmpA beta barrel decreased E. coli resistance to environmental stresses. Moreover, ompA mutants were found to survive much better within brain microvascular endothelial cells than the wild-type strain, supporting that OmpA is a major target in mammalian host cell defense. These results indicated that OmpA plays a vital structural role in E. coli, and suggested that a perfect beta-barrel structure of OmpA is important for outer membrane stability. Based on these results and the published OmpA structural analyses, I propose that OmpA is composed of three functional domains including a hydrophilic extracellular mass, a beta-barrel transmembrane structure, and a peptidoglycan binding domain. PMID- 11906176 TI - Catecholestrogen sulfation: possible role in carcinogenesis. AB - A growing body of evidence supports the hypothesis that estrogens can be carcinogens as a result of their conversion to genotoxins after biotransformation to form the catecholestrogens (CEs) 2-hydroxyestrone (2-OHE1), 2-hydroxyestradiol (2-OHE2), 4-hydroxyestrone (4-OHE1) and 4-hydroxyestradiol (4-OHE2). CEs can then undergo further metabolism to form quinones that interact with DNA to form either stable or depurinating adducts. These events could potentially be interrupted by the sulfate conjugation of both the parent estrogens and/or the CEs. We set out to determine whether CEs can serve as substrates for sulfate conjugation, and-if so-which of the growing family of human sulfotransferase (SULT) isoforms are capable of catalyzing those reactions. We determined apparent K(m) values for 10 recombinant human SULT isoforms, as well as the three most common allozymes for SULT1A1 and SULT1A2, with 2-OHE1, 2-OHE2, 4-OHE1, and 4-OHE2, and with the endogenous estrogens, estrone (E1) and 17beta-estradiol (E2), as substrates. With the exception of SULT1B1, SULT1C1, and SULT4A1, all of the human SULTs studied catalyzed the sulfate conjugation of CEs. SULT1E1 had the lowest apparent K(m) values, 0.31, 0.18, 0.27, and 0.22 microM for 4-OHE1, 4-OHE2, 2-OHE1, and 2-OHE2, respectively. These results demonstrate that SULTs can catalyze the sulfate conjugation of CEs, and they raise the possibility that individual variation in this pathway for estrogen and CE metabolism as a result of common genetic polymorphisms could represent a risk factor for estrogen-dependent carcinogenesis. PMID- 11906177 TI - Exercise-induced transcription of the muscle glucose transporter (GLUT 4) gene. AB - We studied the effects of exercise on GLUT4 gene transcription in several lines of transgenic mice expressing the chloramphenicol acyltransferase (CAT) reporter gene, driven by various lengths of the human GLUT4 promoter (2400, 1600, 895, and 730 bp). In all transgenic lines examined, endogenous GLUT4 mRNA increased in response to exercise (19-90%, P < 0.05). Exercise increased CAT mRNA (51-83%, P < 0.05) in mice when the transgene was driven by at least 895 bp of the promoter but showed no effect in mice in which the transgene was driven by only 730 bp. These results suggest that the exercise-induced increase in the transcriptional activity of the human GLUT4 gene is mediated, at least in part, by element(s) within -895 bp of the promoter. These observations reveal a striking similarity to the time course and regional promoter requirements of AMPK-induced GLUT4 gene expression, providing further evidence that AMPK may be mediating the effects of exercise on GLUT4 expression. PMID- 11906178 TI - Revised structure of the active form of human deoxyribonuclease IIalpha. AB - Deoxyribonuclease IIalpha (DNase IIalpha) is an acid endonuclease found in lysosomes, nuclei, and various secretions. Murine DNase IIalpha is required for digesting the DNA of apoptotic cells after phagocytosis and for correct development and viability. DNase IIalpha purified from porcine spleen was previously shown to contain three peptides, two of which were thiol crosslinked, all derived by processing of a single polypeptide. Commercial bovine protein is consistent with this structure. However, screening of 18 human cell lines failed to demonstrate this processing, rather a 45 kDa protein was consistently observed. Incubation of cells with the N-glycosylation inhibitor tunicamycin resulted in a 37 kDa protein, which is close to the predicted formula weight. The protein also contains at least one thiol crosslink. Similar results were obtained with overexpressed DNase IIalpha. These results suggest that active DNase IIalpha consists of one contiguous polypeptide. We suggest the previous structure reflects proteolysis during protein purification. PMID- 11906179 TI - Promoter analysis of tumor suppressor gene PTEN: identification of minimum promoter region. AB - Mutations of PTEN, a tumor suppressor gene located on chromosome 10, which encodes a protein-tyrosine and lipid-phosphatase, are prevalent in various human cancers, including glioblastoma. Despite extensive characterization of PTEN mutations in human cancers and a relatively good understanding of the molecular roles of PTEN in the control of cellular processes, little is known about modes of PTEN regulation. To understand the regulation of expression of the tumor suppressor gene PTEN, we isolated a 2212 bp fragment from the human BAC clone 46B12 DNA. The 3' end of this fragment starts at the Not I site of -745 relative to the first translation codon ATG (+1) and ends at the Sal I site of -2957 at the 5' end. Using classical 5'RACE and primer extension techniques, nine start sites were observed between -817 and -984 upstream of the ATG start site. We located a 137 bp fragment (-958/-821) as the minimum promoter region using promoter deletion and luciferase assays. A 704 bp fragment (-33/-737) downstream of the 2212 bp fragment was also cloned. As indicated by luciferase assays, the data show that this region possesses no promoter function. Interestingly, a p53 binding sequence is located within the 599 bp fragment (-1344/-745), although p53 expression had a minimal effect on PTEN, demonstrating its insignificant role in PTEN gene expression. PMID- 11906180 TI - Dominant-negative HMGA1 blocks mu enhancer activation through a novel mechanism. AB - The immunoglobulin mu intronic enhancer is a potent B cell-specific transcriptional activator. The enhancer is activated by the appropriate combination of transcription factors, amongst which are ets and bHLH proteins. HMGA1 (formerly HMG-I(Y)) is a demonstrated co-activator of the mu enhancer. HMGA1 functions through direct interaction with PU.1, one of the ets proteins critical for enhancer activation. New data demonstrates dominant negative HMGA1 dramatically decreases enhancer activity in B cells. EMSA analysis demonstrated that DN HMGA1 disrupts established PU.1/mu enhancer binding. Similarly, DN HMGA1 blocks mu enhancer binding by Ets-1. In sharp contrast, DN HMGA1 had no effect on binding activity of the ETS DNA binding domains of either PU.1 or Ets-1, or the bHLH-zip protein TFE3, suggesting specificity. Taken together, the data suggest that DN HMGA1 utilizes a novel mechanism to specifically block interaction between ets proteins and mu enhancer DNA, suggesting DN HMGA1 represents a new, highly specific means of regulating mu enhancer activity. PMID- 11906181 TI - Spontaneous nicking in the nontoxic-nonhemagglutinin component of the Clostridium botulinum toxin complex. AB - The nontoxic-nonhemagglutinin (NTNHA) component, in both isolated form and the neurotoxin (NT)/NTNHA complexed form, was prepared protease-free from toxin complexes produced by Clostridium botulinum type D strain 4947. NTNHA in both preparations was found to be spontaneously converted to the nicked NTNHA form leading to 15- and 115-kDa fragments with the excision of several amino acid residues at specific sites on SDS-PAGE during long-term incubation, while that of the NT/NTNHA/hemagglutinin complexed form remained unnicked single-chain polypeptides under the same conditions. Considering that the NTNHA preparation contained small amounts of the nicked form of NTNHA and the addition of trypsin accelerated the cleavage, it is speculated that a nicked form of NTNHA remaining after the purification and/or NTNHA itself catalyzes the cleavage of intact NTNHA. PMID- 11906183 TI - Role of CD4 hinge region in GP120 utilization by immunoglobulin domain 1. AB - Immunoglobulin-like domain 1 of CD4 (D1-CD4) promotes HIV infection by binding the envelope glycoprotein (ENV) and exposing its coreceptor-binding site. To study CD4-ENV-coreceptor interactions, we characterized hybrid receptors having domains 1 and 2 of CD4 (D1D2-CD4) joined to the N-terminus of chemokine receptors CCR5, CXCR4, CXCR2, and DARC. Hybrid receptors showed conserved ENV-coreceptor specificity in cell-cell fusion assays. Although D1D2-CD4-CCR5 was sufficient to permit ENV-mediated fusion, D1-CD4-CCR5 and human D1/mouse D2-CD4-CCR5 lacked CD4 function and binding to a neutralizing antibody mapped to D1-CD4. Chimeric D1D2 CD4 joined to CCR5 revealed that the C-terminal 20 residues of human D2-CD4 are required for efficient ENV-mediated fusion. Mutagenesis of hybrid receptors showed the importance of residues forming D1-D2 CD4 interdomain contacts and hinge region proximal residues. Mutagenesis of WT human CD4 confirmed that residues forming D1-D2 interdomain contacts and hinge-region proximal residues contribute positively to CD4 activity in the full-length receptor. PMID- 11906182 TI - Production of antiviral and antitumor proteins MAP30 and GAP31 in cucurbits using the plant virus vector ZYMV-AGII. AB - ZYMV-AGII (zucchini yellow mosaic virus-AGII) is a recombinant nonpathogenic potyvirus-based vector system for the expression of foreign genes in cucurbit plants and their edible fruits, including squash, cucumber, melon, watermelon, and pumpkin. MAP30 (Momordica anti-HIV protein, 30 kDa) and GAP31 (Gelonium anti HIV protein 31 kDa) are multifunctional plant proteins with activity against HIV 1 virus. These proteins are also effective against other viruses, tumor cells, and microbes. We report here the production and characterization of biologically active MAP30 and GAP31 in squash plant by expression of their genes using the ZYMV-AGII vector. Recombinant expressed MAP30 and GAP31 exhibit comparable antiviral, antitumor, and antimicrobial activities as their counterparts from their original plant sources, with EC(50)s in the ranges of 0.2-0.3 nM for HIV-1. These results demonstrate for the first time the amplification and production of therapeutic proteins, MAP30 and GAP31, in common vegetables. This provides valuable alternative food sources of these antiviral, antitumor, and antimicrobial agents for therapeutic applications. PMID- 11906185 TI - Purification and characterization of two active derivatives of recombinant YplA, a secreted phospholipase from Yersinia entercolitica. AB - The virulence-associated phospholipase of Yersinia enterocolitica (YplA), which is secreted by a flagellar type III secretion system, was cloned and purified for structure-function analysis using a His(6)-tag expression system. Two versions of YplA have been proposed on the basis of two potential initiating methionine residues. The longer derivative possesses 59 additional amino acids at its N terminus and appears to represent the native form of YplA; however, the shorter recombinant protein possesses enhanced activity in vitro. Both recombinant YplA derivatives are highly active as type-A(2) phospholipases and possess similar physical properties. Based on type III secretion substrates from other gram negative bacteria, the N-terminus of YplA is probably required as a secretion signal; however, differences in the time-based activity of these two recombinant enzymes, the N-terminus of YplA may also have a regulatory function. PMID- 11906186 TI - The mechanism of salivary amylase hydrolysis: role of residues at subsite S2'. AB - Hydrolysis of starch or oligosaccharides by mammalian amylases, in general, results in maltose as the leaving group. The active site of these amylases harbors three aromatic residues Trp59, Tyr62, and Tyr151, which provide stacking interactions to the bound glucose moieties. We hypothesized that Tyr151, located at the S2' subsite, may influence the size of the leaving group. Therefore, using a baculovirus expression system, we generated a mutant Y151M in which the tyrosine at position 151 of human salivary amylase is replaced by a methionine. The specific activity, K(m), rate of hydrolysis, and the product distribution for Y151M were distinctly different from those of the wild-type enzyme using starch and oligosaccharides as substrates. The mutant enzyme Y151M consistently produced glucose as the minimal leaving group and exhibited a twofold increase in K(m). These results suggest that the stacking interaction at subsite S2' in the wild type plays a role in hydrolysis. PMID- 11906184 TI - Bisphenol A induces apoptosis and G2-to-M arrest of ovarian granulosa cells. AB - We investigated the impact of bisphenol A (BPA) on murine ovarian granulosa cells. Ovarian granulosa cells were cultured with 100 fM to 100 microM BPA for 24 h to 72 h. BPA decreased granulosa cell viability in a dose- and time-dependent manner. The lowest concentration that induced a significant decrease was 100 pM (89.2 +/- 4.0% of the control). TUNEL analysis demonstrated that treatment with BPA increased apoptosis of granulosa cells in a dose- and time-dependent manner. In addition, flow cytometry analyses revealed that treatment with BPA resulted in G2-to-M arrest, which was most prominent at 48 h. BPA increased the expression of Bax and concomitantly decreased the expression of Bcl2 at both protein and mRNA levels of granulosa cells. These findings suggest that low, presumably environmentally relevant doses of BPA, decrease the viability of granulosa cells by inducing apoptosis and G2-to-M arrest. Up-regulation of Bax and down regulation of Bcl2 were suggested to be involved in this apoptotic effect. PMID- 11906187 TI - Inhibited activities in CCAAT/enhancer-binding protein, activating protein-1 and cyclins after hepatectomy in rats with thioacetamide-induced liver cirrhosis. AB - Transcriptional activation of nuclear factor (NF)-kappaB, signal transducers and activators of transcription (STAT) 3, activating protein (AP)-1 and CCAAT/enhancer-binding protein (C/EBP) plays an important role in liver regeneration by modulating cell cycle regulators. The regeneration of cirrhotic liver after hepatectomy is inhibited despite intact expression of growth factors. To elucidate the mechanism involved, regeneration responses in growth factor receptors, transcription factors, and cell cycle regulators after two-thirds hepatectomy were compared between rats with thioacetamide-induced cirrhotic and normal liver. The expression of c-met and epidermal growth factor receptor analyzed by RT-PCR and immunohistochemistry did not differ between the two groups. The activities of C/EBP and AP-1 evaluated by electrophoretic mobility shift assay were significantly inhibited in the cirrhotic group compared with those in the control group, but not those of NF-kappaB and STAT3. The expression of cyclin-D1, -E, and -A assessed by Western blot analysis was significantly decreased in the cirrhotic group compared with the control group. The level in p21(Cip1) or p27(Kip1) did not differ between the two groups. The liver regeneration estimated by the rates of [(3)H]thymidine incorporation into DNA and staining of proliferating cell nuclear antigen was significantly lower in the cirrhotic rats than in the controls. In conclusion, downregulation of cyclin -D1, -E, and -A expression, which may be induced by impaired activities of C/EBP and AP-1, is responsible for the decreased regenerative capacity of cirrhotic liver after partial hepatectomy. PMID- 11906188 TI - Effects of aging on growth factors gene and protein expression in the dorsal and ventral lobes of rat prostate. AB - We hypothesize that various growth factors and their receptors gene and protein are modulated in dorsal and ventral lobes of aging prostate. To test this hypothesis, TGFbeta1, TGFbeta2 TGFbeta3, TGFbetaR-I, TGFbetaR-II, TGFalpha, EGF, EGFR, KGF and KGFR gene and protein expression were analyzed in dorsal and ventral lobes of aging rat prostates (1, 3, 6, 9, 12, 18, 24, and 28/30 months). KGF gene expression was very weak or absent in 1, 3, and 6 month old rat dorsal and ventral lobes of prostate whereas it re-expressed in 9, 12, 18, 24 and 30 month old rat prostate. All growth factors and their receptors expect KGF and EGFR were mainly localized in epithelium of ventral and dorsal lobes of aging rat prostates. EGF, TGFalpha, TGFbeta1, and TGFbetaR-I protein expression was lacking in stroma of dorsal and ventral lobes of 1, 3, 6, 9, 12/18 months old rat prostates. However, EGF, TGFbeta1 and TGFbetaR-I proteins re-expressed in stroma of 24 and 28 months old rat prostates. KGF protein expression was lacking in epithelium of dorsal and ventral lobes of all aging rat prostates. This is the first report to demonstrate differential gene and protein expression of growth factors in dorsal and ventral lobes is associated with aging rat prostate, suggesting their role in pathogenesis of prostatic diseases with aging. PMID- 11906189 TI - Identification of a defect in the UGT1A1 gene promoter and its association with hyperbilirubinemia. AB - The UDP-glucuronosyltransferase UGT1A1 plays a critical role in the detoxification of potentially neurotoxic bilirubin by conjugating it with glucuronic acid. We identified a polymorphism that results in a T to G substitution at nucleotide number -3263 of the phenobarbital-responsive enhancer module of the UGT1A1 gene, thereby significantly decreasing transcriptional activity as indicated by the luciferase-reporter assay. At least one T-3263G allele was found in 21 of 25 subjects with mild hyperbilirubinemia (Gilbert's syndrome); this frequency (0.58) was significantly higher than that in normobilirubinemic controls (0.17; n = 8 of 27). Homozygous mutations in the TATA element (A[TA](7)TAA) or at nucleotide 211 of exon 1 (G to A substitution) were found in 5 and 2 of the hyperbilirubinemic group, respectively, while 12 of these subjects were double heterozygotes for the T-3263G and G211A mutations. Plasma total bilirubin levels in these double heterozygotes were significantly higher than those in control subjects carrying one or other of these mutations singly, indicating that compound heterozygous mutations may result in more strongly reduced UGT1A1 activity. Our results indicate that homozygosity and compound heterozygosity for mutations in the UGT1A1 gene promoter (T-3263G and A[TA](7)TAA) and/or exon 1 of the gene (G211A) could explain the hyperbilirubinemia seen in the majority of individuals with Gilbert's syndrome. PMID- 11906190 TI - Expression profile analysis of colon cancer cells in response to sulindac or aspirin. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have a preventive effect against colorectal cancer. Although inhibition of cyclooxygenase-2 plays a crucial role in the suppression of tumors, precise mechanisms of their action remain to be disclosed. To identify genes involved in the growth-suppressive effect of NSAIDs, we utilized cDNA microarray containing 23,040 genes and analyzed time-dependent alteration of gene expression in response to sulindac or aspirin in NSAIDs sensitive SW480 and SW948 colon-cancer cells as well as in relatively resistant SNU-C4 cells. Consequently we identified 112 genes with commonly altered expression by sulindac and 176 with commonly altered expression by aspirin in the three lines. Addition of sulindac and that of aspirin altered expression levels of 130 and 140 genes, respectively, in SW480 and SW948 cells but not in SNU-C4 cells. These data may lead to a better understanding of growth-suppressive effects on colonic epithelium, and may provide clues for identifying novel therapeutic and/or preventive molecular targets of colon cancer. PMID- 11906191 TI - Secondary-site mutation restores the transport defect caused by the transmembrane domain mutation of the xenobiotic transporter MexB in Pseudomonas aeruginosa. AB - It has been suggested that the MexB subunit of the MexAB-OprM efflux transporter of Pseudomonas aeruginosa exports xenobiotics in an energy-dependent manner. To investigate the role of the transmembrane segments (TMS) of MexB in the transporter activity, we isolated 24 spontaneous mutants showing hypersusceptibility to antibiotics. Among them, three mutations were located at TMS-3, TMS-4, and TMS-10 having amino acid substitution Leu376vPro, Gly397vVal, and Val928vGly, respectively. A secondary mutation, which suppressed the defect caused by the Val928vGly mutation in TMS-10, was found at the 403rd amino acid residue in TMS-4 with a change of glycine to serine, suggesting that TMS-4 and TMS-10 may be in close proximity. This result provided strong support for the recent notion that negatively charged residues in TMS-4 might form a salt-bridge with a positive charge in TMS-10 (Guan, L., and Nakae, T. (2001) J. Bacteriol. 183, 1734-1739). The transporter function impaired by the Gly397vVal mutation in TMS-4 was recovered by the secondary mutation, Gln998vHis, in the loop between TMS-11 and TMS-12, thereby suggesting that TMS-4 and TMS-11 or TMS-12 might also be in close proximity. Thus, it is most likely that TMS-4, TMS-10, and TMS-11 or TMS-12 are packed close three dimensionally. PMID- 11906192 TI - Mechanistic studies on AMD6221: a ruthenium-based nitric oxide scavenger. AB - Nitric oxide is a mediator of many disease states. Previous studies have demonstrated that ruthenium(III) polyaminocarboxylates can react with NO to form stable complexes reducing the levels of nitrite in the culture medium of stimulated RAW264 macrophages and reverse the NO-mediated hypotension in animal models of septic shock. It was necessary to confirm that these observations were due to NO scavenging and not inhibition of the NO metabolic pathway. Using RAW264 cells it was confirmed that [Ru(H(3)dtpa)(Cl)] (AMD6221) was neither acting at the level of iNOS induction, nor as an inhibitor of iNOS by measuring iNOS mRNA by RT-PCR and protein by Western blot and enzyme activity. Using HPLC, the nitrosyl adduct of reaction of AMD6221, [Ru(H(2)dtpa)NO], was identified in the medium of stimulated RAW264 cells co-incubated with AMD6221, concomitant with a stoichiometric reduction in nitrite/nitrate levels, thus confirming that the ruthenium(III) polyaminocarboxylates exert their pharmacological effect by scavenging NO. PMID- 11906193 TI - Saccharomyces cerevisiae resistance to chlorinated phenoxyacetic acid herbicides involves Pdr1p-mediated transcriptional activation of TPO1 and PDR5 genes. AB - The transcription regulator Pdr1p is a determinant of Saccharomyces cerevisiae resistance to 2-methyl-4-chlorophenoxyacetic acid (MCPA) and 2,4 dichlorophenoxyacetic acid (2,4-D). The Pdr1p-regulated genes, TPO1 and PDR5, encoding putative multidrug transporters belonging to the major facilitator superfamily (MFS) and to the ATP-binding cassette (ABC) superfamily, respectively, are required for yeast resistance to sudden exposure to these herbicides. A rapid and transient activation of TPO1 (sixfold) and PDR5 (twofold) transcription takes place during the adaptation period preceding cell division under MCPA or 2,4-D moderate stress. These activations are mediated by both Pdr1p and Pdr3p and, as soon as adapted cells start duplication under herbicide stress, mRNA levels are drastically reduced to basal values. The longer duration of the adaptation period, observed for the Delta(pdr1) population, may involve the abolishment of the Pdr1p-mediated transcriptional activation of TPO1 and PDR5 genes, whose expression is critical to surpass the viability loss during the initial period of adaptation to the herbicides. PMID- 11906194 TI - TNFalpha production in whole blood cultures from healthy individuals. AB - Tumor necrosis factor alpha (TNFalpha) is a major mediator of inflammatory responses and also plays a prominent role in bridging the innate and adaptive phases of immunity. In the present work we attempted to study TNFalpha production in endotoxin-stimulated blood of healthy individuals, and the inter-individual variability in TNFalpha production. For this study, we used diluted whole blood stimulated with lipopolysaccharide (LPS). The levels of the pro-inflammatory cytokine TNFalpha were measured by ELISA and by the L929 cytotoxicity bioassay in 16 and 18 healthy donors, respectively. There were highly significant inter individual variations in the induced TNFalpha production. It is worth noting that there was no difference in sensitivity between ELISA and the cytotoxicity L929 bioassay. We concluded that whole blood culture is a sensitive method to determine the pro-inflammatory cytokine production in response to endotoxin stimuli in a relevant physiologic milieu. Our data indicate that this method provides appropriate information about the state of cellular immunity of the individual. PMID- 11906195 TI - Cellular defense against UVB-induced phototoxicity by cytosolic NADP(+)-dependent isocitrate dehydrogenase. AB - Ultraviolet (UV) radiation is known as a major cause of skin photoaging and photocarcinogenesis. Many harmful effects of UV radiation are associated with the generation of reactive oxygen species. Recently, we have shown that NADP(+) dependent isocitrate dehydrogenase is involved in the supply of NADPH needed for GSH production against cellular oxidative damage. In this study we investigated the role of cytosolic form of NADP(+)-dependent isocitrate dehydrogenase (IDPc) against UV radiation-induced cytotoxicity by comparing the relative degree of cellular responses in three different NIH3T3 cells with stable transfection with the cDNA for mouse IDPc in sense and antisense orientations, where IDPc activities were 2.3-fold higher and 39% lower, respectively, than that in the parental cells carrying the vector alone. Upon exposure to UVB (312 nm), the cells with low levels of IDPc became more sensitive to cell killing. Lipid peroxidation, protein oxidation, oxidative DNA damage, and intracellular peroxide generation were higher in the cell-line expressing the lower level of IDPc. However, the cells with the highly overexpressed IDPc exhibited enhanced resistance against UV radiation, compared to the control cells. The data indicate that IDPc plays an important role in cellular defense against UV radiation induced oxidative injury. PMID- 11906196 TI - Annexin-V binds to the intracellular part of the beta(5) integrin receptor subunit. AB - Bovine lactadherin binds to the alpha(v)beta(3) and alpha(v)beta(5) integrins in an RGD-dependent manner and also to anionic phospholipids. During the affinity purification of lactadherin binding receptors, a 35-kDa protein persistently coeluted with the alpha(v)beta(5) integrin receptor. Subsequently, peptide mapping, amino acid sequencing, and mass spectrometry analysis identified this protein as bovine annexin-V. Annexin-V accompanied the integrin receptor eluted with either RGD peptide or with EDTA suggesting that annexin-V bound specifically to the alpha(v)beta(5) integrin. To further investigate this putative interaction of annexin-V with the alpha(v)beta(5) integrin receptor, human annexin-V and intracellular domains of the human alpha(v)beta(5) integrin subunits were used in ligand blotting assays. Radiolabeled annexin-V showed weak binding to the intracellular part of beta(5) integrin subunit. However, by adding the aminophospholipid, phosphatidyl serine, the interaction with the beta(5) cytoplasmic peptide was enhanced many fold. Furthermore, the interaction was shown to be independent of phosphorylation, as annexin-V bound to unphosphorylated beta(5) peptide at a similar level to the phosphorylated peptide. Since binding of annexin-V to the alpha(v) integrin subunit tail was not detected, annexin-V was shown to associate specifically with the beta(5) cytoplasmic tail. Together these findings suggest a novel link between annexins and the integrin receptor family. PMID- 11906197 TI - The FMO2 gene of laboratory rats, as in most humans, encodes a truncated protein. AB - We describe the isolation and characterization of cDNAs for FMO2 from the laboratory rat. In contrast to FMO2 in other animals, each of which contain 535 amino acid residues, analysis of the sequence of the cDNAs and of a section of the corresponding gene revealed that the ORF of the laboratory rat FMO2 encodes a polypeptide of only 432 residues. This truncated protein is due to the presence of a double deletion corresponding to 1263 and 1264 nucleotides of the orthologous FMO2 cDNAs. This double deletion provokes a frame-shift, with the appearance of a premature stop codon in position 1297-1299. By Northern blotting, the probe for FMO2 hybridized a 2.5-kb transcript in lung and kidney samples only. Heterologous expression of the cDNA revealed that the truncated protein was catalytically inactive. By Western blotting, FMO2 was faintly detected at approximately 50 kDa in laboratory rat lung. PMID- 11906198 TI - Early developing embryos affect the gene expression patterns in the mouse oviduct. AB - Fertilization and development of mouse embryos occur in the ampullae of oviduct. We hypothesize that fetal-maternal communication exists in the preimplantation period, allowing optimal development of embryos. It is known that embryotrophic factors from oviduct affect the development of embryos. Although embryos affect their own transport in the oviduct, the mechanism of action is unknown. As a step toward understanding the action of embryos on oviductal physiology, we adopted suppression subtractive hybridization (SSH) to compare the gene expression in the mouse oviduct containing early embryos with that of oviduct containing oocytes. Ten to twelve 1-cell mouse embryos were transferred to one oviduct of a foster mother and similar number of oocytes were transferred to the contralateral oviduct. The animals were sacrificed after 48 h and their oviducts were excised for mRNA study. Using SSH, we screened out 250 putative positive clones from the subtracted embryo-containing oviduct library and 97 of them were screened positive by reverse dot-blot analysis. DNA sequence analysis identified genes that shared high homology with sequences in GenBank/EMBL database with unknown functions. Overall, 13 of the 90 high-quality sequences (14%) were homologous to 6 different genes previously described. Reverse Northern analysis confirmed that the expression of these genes were higher in the embryo-containing oviduct than in the oocyte-containing oviduct. About 12% of these clones (11/90) were novel. This article is the first to report identification of genes in the oviduct that are upregulated in the presence of embryos during the preimplantation period. PMID- 11906199 TI - Generation of amyloid beta protein from a presenilin-1 and betaAPP complex. AB - Presenilin-1 (PS1) is a causative gene in early onset familial Alzheimer's disease (FAD). FAD-linked mutant PS1s significantly increased Abeta40 and Abeta42(43) levels (P < 0.001) and decreased the production of an 11.4 kD (beta stub) and an 8.7 kD (alpha-stub) carboxyl-terminal fragment of amyloid beta precursor protein (betaAPP-CTFs) (P < 0.01). In the 2% CHAPS extracted lysates, the complex containing the amino-terminal fragment of PS1 (PS1-NTF), the carboxyl terminal fragments of PS1 (PS1-CTF), and betaAPP-CTFs was identified. Incubation of this isolated complex at pH 6.4 showed the direct generation of Abeta40 and gamma-stub from this complex. This reaction was inhibited by a gamma-secretase inhibitor. The degrading rate of a co-precipitated beta-stub was facilitated under the presence of FAD-linked mutant PS1s. This findings suggest that the direct generation of Abeta from the complex may play an important role in the pathogenesis of Alzheimer's disease. PMID- 11906201 TI - The "Frontiers" Spirit of William F. Ganong and Luciano Martini. PMID- 11906200 TI - Melittin-GM1 interaction: a model for a side-by-side complex. AB - We report here the interaction of melittin with ganglioside GM1 by steady-state fluorescence, one-dimensional (1)H NMR spectroscopy and molecular modeling. In the presence of GM1 the emission maximum of melittin is blue shifted and fluorescence quenching efficiencies of iodide and acrylamide are substantially reduced, indicating a shielding of tryptophan of melittin from aqueous environment. Significant line broadening of NMR resonances of melittin, suggestive of motional restriction, is observed. Molecular modeling indicates a melittin-GM1 complex with N-terminal hydrophobic stretch of melittin associating with the ceramide tail and C-terminal hydrophilic end of melittin having favorable electrostatic interaction with the carbohydrate head group of GM1. PMID- 11906202 TI - The neuroanatomical axis for control of energy balance. AB - The hypothalamic feeding-center model, articulated in the 1950s, held that the hypothalamus contains the interoceptors sensitive to blood-borne correlates of available or stored fuels as well as the integrative substrates that process metabolic and visceral afferent signals and issue commands to brainstem mechanisms for the production of ingestive behavior. A number of findings reviewed here, however, indicate that sensory and integrative functions are distributed across a central control axis that includes critical substrates in the basal forebrain as well as in the caudal brainstem. First, the interoceptors relevant to energy balance are distributed more widely than had been previously thought, with a prominent brainstem complement of leptin and insulin receptors, glucose-sensing mechanisms, and neuropeptide mediators. The physiological relevance of this multiple representation is suggested by the demonstration that similar behavioral effects can be obtained independently by stimulation of respective forebrain and brainstem subpopulations of the same receptor types (e.g., leptin, CRH, and melanocortin). The classical hypothalamic model is also challenged by the integrative achievements of the chronically maintained, supracollicular decerebrate rat. Decerebrate and neurologically intact rats show similar discriminative responses to taste stimuli and are similarly sensitive to intake-inhibitory feedback from the gut. Thus, the caudal brainstem, in neural isolation from forebrain influence, is sufficient to mediate ingestive responses to a range of visceral afferent signals. The decerebrate rat, however, does not show a hyperphagic response to food deprivation, suggesting that interactions between forebrain and brainstem are necessary for the behavioral response to systemic/ metabolic correlates of deprivation in the neurologically intact rat. At the same time, however, there is evidence suggesting that hypothalamic neuroendocrine responses to fasting depend on pathways ascending from brainstem. Results reviewed are consistent with a distributionist (as opposed to hierarchical) model for the control of energy balance that emphasizes: (i) control mechanisms endemic to hypothalamus and brainstem that drive their unique effector systems on the basis of local interoceptive, and in the brainstem case, visceral, afferent inputs and (ii) a set of uni- and bidirectional interactions that coordinate adaptive neuroendocrine, autonomic, and behavioral responses to changes in metabolic status. PMID- 11906203 TI - Diverse actions of ovarian steroids in the serotonin neural system. AB - All of the serotonin-producing neurons of the mammalian brain are located in 10 nuclei in the mid- and hindbrain regions. The cells of the rostal nuclei project to almost every area of the forebrain and regulate diverse neural processes from higher order functions in the prefrontal cortex such as integrative cognition and memory, to limbic system control of arousal and mood, to diencephalic functions such as pituitary hormone secretion, satiety, and sexual behavior. The more caudal serotonin neurons project to the spinal cord and interact with numerous autonomic and sensory systems. All of these neural functions are sensitive to the presence or absence of the ovarian hormones, estrogen and progesterone. We have shown that serotonin neurons in nonhuman primates contain estrogen receptor beta and progestin receptors. Thus, they are targets for ovarian steroids which in turn modify gene expression. Any change in serotoninergic neural function could be manifested by a change in any of the projection target systems and in this manner, serotonin neurons integrate steroid hormone information and partially transduce their action in the CNS. This article reviews the work conducted in this laboratory on the actions of estrogens and progestins in the serotonin neural system of nonhuman primates. Comparisons to results obtained in other laboratory animal models are made when available and limited clinical data are referenced. The ability of estrogens and progestins to alter the function of the serotonin neural system at various levels provides a cellular mechanism whereby ovarian hormones can impact cognition, mood or arousal, hormone secretion, pain, and other neural circuits. PMID- 11906204 TI - Oxytocin-secreting neurons: A physiological model of morphological neuronal and glial plasticity in the adult hypothalamus. AB - Oxytocin-secreting neurons of the hypothalamoneurohypophysial system undergo reversible morphological changes whenever they are strongly stimulated. In the hypothalamus, such structural plasticity is represented by modifications in the size and shape of their somata and dendrites, in the extent to which their surfaces are covered by glia, and in the density of their synapses. In the neurohypophysis, there is a parallel reduction in glial (pituicyte) coverage of their axons together, with retraction of pituicyte processes from the perivascular basal lamina and an increase in the number and size of their terminals. These changes occur rapidly, within a few hours. On the other hand, the system returns to its prestimulated condition on arrest of stimulation at a rate that depends on the length of time it has remained activated. Such neuronal glial changes have several functional consequences. In the hypothalamic nuclei, reduction in astrocytic coverage of oxytocinergic neurons and their synapses modifies extracellular ionic homeostasis and glutamate clearance and, therefore, their overall excitability. Since it results in extensive dendritic bundling, it may also lead to ephaptic interactions and may facilitate dendritic electrotonic coupling. A most important indirect effect may be to permit synaptic remodeling that occurs concomitantly and that results in significant increases in the number of excitatory and inhibitory synapses driving their activity. In the stimulated neurohypophysis, glial retraction results in increased levels of extracellular K+ which can enhance neurohormone release while an enlarged neurovascular contact zone may facilitate diffusion of neurohormone into the circulation. Ongoing work aims to unravel the cell mechanisms and factors underlying such plasticity and has revealed that neurons and glia of the hypothalamoneurohypophysial system continue to express juvenile molecular features associated with similar neuronglial interactions and synaptic events during development and regeneration. They include strong expression of cell surface adhesion molecules like F3/contactin and polysialylated neural cell adhesion molecule, extracellular matrix glycoproteins like tenascin C, and cytoskeletal proteins like vimentin and microtubule-associated protein 1D. Some of these molecules reach the cell surface constitutively while others follow the activity-dependent regulated pathway. We consider many of these molecular features permissive, allowing oxytocin neurons and their glia to undergo morphological remodeling throughout life, provided the proper stimulus intervenes. In the hypothalamic nuclei, one such stimulus is centrally released oxytocin; in the neurohypophysis, an adrenergic, cAMP-mediated mechanism appears responsible. PMID- 11906205 TI - TGFbeta1 modulates the phenotype of Schwann cells at the transcriptional level. AB - We have examined the effects of transforming growth factor beta1 (TGFbeta1) on gene expression in cultured rat Schwann cells (SCs). TGFbeta1 decreased the steady-state mRNA levels of several genes that are expressed by myelinating SCs but had varied effects on the mRNA levels of NCAM, L1, GAP-43, and p75-genes that are expressed by denervated and nonmyelinating SCs. TGFbeta1 antagonized the effects of forskolin on the mRNA levels of the transcription factors Oct-6/tst 1/SCIP and Krox20. Transcriptional run-off analysis demonstrated that the effects of TGFbeta1 on gene expression occur at least in part at the level of transcription. Thus, TGFbeta1 suppresses the expression of genes that characterize the different phenotypes of SCs, and these changes occur at least in part at a transcriptional level. PMID- 11906206 TI - Sonic hedgehog is a potent inducer of rat oligodendrocyte development from cortical precursors in vitro. AB - Sonic Hedgehog (Shh) induces oligodendrocyte development in the ventral neural tube and telencephalon but its role in oligodendrocyte generation in dorsal telencephalon is debated. Transcripts for Shh and its receptor complex were detected in subventricular zone and neocortex from E17 to birth. As Shh is not yet expressed in E15 neocortex, we grew E15 cortical precursors (CP) into neurospheres in the presence of recombinant Octyl-Shh (O-Shh). After sphere adhesion and removal of O-Shh, enhanced neurite outgrowth and cell migration were already observed at 3 h. Three days after O-Shh treatment, oligodendrocyte progenitors (OP) emerged and continued to increase in number for 7 days while the ratio of neuronal cells decreased compared to control. Shh selectively triggered mitosis of OP but not neuronal progenitors and enhanced growth of neonatal OP. Thus Shh in E15-17 embryonic neocortex can signal CP to adopt an oligodendrocyte fate and favors expansion of this lineage. PMID- 11906207 TI - Thyroid hormone-dependent regulation of Talpha1 alpha-tubulin during brain development. AB - Thyroid hormone (T3) is essential for brain development and most of its actions are exerted at the gene expression level after interaction with nuclear receptors. In particular, genes encoding cytoskeletal proteins are influenced by the thyroidal status. Thyroid hormone is involved in the normal downregulation of the Talpha1 alpha-tubulin gene during postnatal growth. The action of T3 on Talpha1 tubulin expression is complex and is exerted at least at two levels. In cultured cells, T3 induces a transient and fast decrease of Talpha1 mRNA concentration. This effect is enhanced when transcription is blocked by actinomycin D, suggesting that T3 increases mRNA degradation. In transgenic animals T3 affects the expression of beta-galactosidase under control of the Talpha1 promoter in the same way as the endogenous gene, supporting an effect mediated through the Talpha1 promoter. However, the Talpha1 promoter is not regulated by T3 in transfected cells and, therefore, the effects of the hormone in vivo are likely to be indirect. It is concluded that regulation of Talpha1 alpha-tubulin by thyroid hormone is the result of multiple influences including effects on mRNA half life and indirect effects at the promoter level. PMID- 11906208 TI - Laminets: laminin- and netrin-related genes expressed in distinct neuronal subsets. AB - Laminins and netrins are families of related secreted proteins known to play critical roles in guiding the growth of peripheral and central axons, respectively. Here we report the identification of two novel cell surface glycoproteins that we name laminets because they resemble both laminins and netrins. Laminet-1 and -2 are selectively expressed in neurons, each in a distinct subset that includes populations in forebrain, midbrain, hindbrain, spinal cord, and spinal ganglia. In several forebrain regions, including main relays of the central olfactory pathway, laminet-1 and -2 are expressed in nonoverlapping neuronal subsets. Both laminets are subject to alternative splicing which, in the case of laminet-1, generates at least 10 distinct isoforms, each of which contains a unique combination of potential binding sites for ligands or counterreceptors. Their complex patterns of distribution and isoform diversity, along with their homology to known axon guidance molecules, suggest that laminets contribute to the patterning of neuronal connections. PMID- 11906209 TI - The cyclin-dependent kinase inhibitors p19(Ink4d) and p27(Kip1) are coexpressed in select retinal cells and act cooperatively to control cell cycle exit. AB - Cyclin-dependent kinase inhibitors (cdki's), including p19(Ink4d) and p27(Kip1), mediate exit from the cell cycle. To determine the function of these cdki's in regulating neurogenesis, we examined retina from wild-type, Ink4d-null, and Ink4d/Kip1-double null animals. Ink4d was expressed in progenitors and select neurons in the mature retina. Ink4d-null retina showed an extended period of proliferation, followed by apoptosis. Colabeling for p19(Ink4d) and p27(Kip1) revealed that a subpopulation of cells expressed both inhibitors. Deletion of Ink4d and Kip1 resulted in continued proliferation that was synergistic. This hyperproliferation led to an increase in number of horizontal cells and differentiated neurons reentering the cell cycle. Deletion of Ink4d and Kip1 also exacerbated the retinal dysplasia observed in Kip1-null mice, which was shown to be partly dependent on p53. These data indicate that select retinal cells express both p19(Ink4d) and p27(Kip1) and that they act cooperatively to ensure cell cycle exit. PMID- 11906210 TI - DN-cadherin is required for spatial arrangement of nerve terminals and ultrastructural organization of synapses. AB - We studied roles of DN-cadherin, the Drosophila major neuronal cadherin, in neuronal connections in the visual system. In DN-cadherin mutants, axon terminals of a large subset of photoreceptor cells reached and associated with their target interneurons, but their characteristic spatial arrangement was disrupted as synaptogenesis proceeded. Although synapses were formed at contact sites between the axon terminals and target neurons, underlying cytoplasmic structures were not fully specialized at both pre- and postsynaptic terminals and synaptic vesicles appeared to accumulate at the presynapses. These results suggest that the cadherin adhesion system is required for interaction between pre- and postsynaptic terminals and for generation of the mature synaptic structures. PMID- 11906211 TI - Long-term expression of beta-glucuronidase by genetically modified human neural progenitor cells grafted into the mouse central nervous system. AB - Mucopolysaccharidosis type VII (MPS VII) is an inherited disease caused by beta glucuronidase (beta-glu) deficiency. This deficiency results in the lysosomal accumulation of glycosaminoglycans in all tissues and affects a wide range of organs, including the central nervous system (CNS). Gene transfer is a promising approach to therapy for MPS VII because it allows extensive delivery of the enzyme to the affected tissues. We studied neurotransplantation of primary human cells to supply beta-glucuronidase to the CNS. Human neural progenitor cells (HNPC) were amplified and cotransduced with two lentiviral vectors, one encoding the green fluorescent protein and the other the human beta-glu. We show that these cells strongly expressed both transgenes in culture. When grafted into the mouse striatum, HNPC differentiated into neurons and astrocytes and expressed the two transgenes for at least 6 months. This study therefore paves the way for the treatment of MPS VII by long-term delivery of the appropriate enzyme. PMID- 11906212 TI - tipE regulates Na+-dependent repetitive firing in Drosophila neurons. AB - The tipE gene, originally identified by a temperature-sensitive paralytic mutation in Drosophila, encodes a transmembrane protein that dramatically influences sodium channel expression in Xenopus oocytes. There is evidence that tipE also modulates sodium channel expression in the fly; however, its role in regulating neuronal excitability remains unclear. Here we report that the majority of neurons in both wild-type and tipE mutant (tipE-) embryo cultures fire sodium-dependent action potentials in response to depolarizing current injection. However, the percentage of tipE- neurons capable of firing repetitively during a sustained depolarization is significantly reduced. Expression of a tipE+ transgene, in tipE- neurons, restores repetitive firing to wild-type levels. Analysis of underlying currents reveals a slower rate of repolarization-dependent recovery of voltage-gated sodium currents during repeated activation in tipE- neurons. This phenotype is also rescued by expression of the tipE+ transgene. These data demonstrate that tipE regulates sodium-dependent repetitive firing and recovery of sodium currents during repeated activation. Furthermore, the duration of the interstimulus interval necessary to fire a second full-sized action potential is significantly longer in single- versus multiple-spiking transgenic neurons, suggesting that a slow rate of recovery of sodium currents contributes to the decrease in repetitive firing in tipE- neurons. PMID- 11906213 TI - Gas1 is induced during and participates in excitotoxic neuronal death. AB - We have performed differential screening to identify genes participating in NMDA induced neuronal death. The gas1 (growth arrest-specific gene 1) gene, whose product is known to inhibit cell cycle progression, was induced in cultured corticohippocampal neurons committed to die after a brief exposure to NMDA. Overexpression of Gas1 in cultured hippocampal neurons and in human neuroblastoma NB69 cells produced a marked reduction in the number of viable cells. Furthermore, gas1 antisense oligodeoxynucleotide or antisense mRNA protected hippocampal neurons or NB69 cells from neuronal death. Importantly, Gas1-induced neuronal death was attenuated by coexpression of the human Bcl-2 protein or the baculoviral caspase inhibitor OpIAP2. While Gas1 does not directly interact with Bcl-2, OpIAP2 coimmunoprecipitates with Gas1. In addition, induction of gas1 also occurred in rat brain in two models of excitotoxicity: delayed neuronal death after intraperitoneal kainate injection and neuronal death in hippocampal slices after ischemia. These results indicate that Gas1 is induced by activation of glutamate receptors and is part of the gene expression program directing neuronal death after mild excitotoxic insults. PMID- 11906214 TI - Regulated nuclear trafficking of the homeodomain protein otx1 in cortical neurons. AB - Otx1 is a homeodomain protein required for axon refinement by layer 5 neurons in developing cerebral cortex. Otx1 localizes to the cytoplasm of progenitor cells in the rat ventricular zone, and remains cytoplasmic as neurons migrate and begin to differentiate. Nuclear translocation occurs during the first week of postnatal life, when layer 5 neurons begin pruning their long-distance axonal projections. Deletion analysis reveals that Otx1 is imported actively into cell nuclei, that the N-terminus of Otx1 is necessary for nuclear import, and that a putative nuclear localization sequence within this domain is sufficient to direct nuclear import in a variety of cell lines. In contrast, GFP-Otx1 fusion proteins that contain the N-terminus are retained in the cytoplasm of cortical progenitor cells, mimicking the distribution of Otx1 in vivo. These results suggest that ventricular cells actively sequester Otx1 in the cytoplasm, either by preventing nuclear import or by promoting a balance of export over import signals. PMID- 11906215 TI - AMPA receptor-mediated modulation of inward rectifier K+ channels in astrocytes of mouse hippocampus. AB - Astrocytes and neurons are tightly associated and recent data suggest a direct signaling between neuronal and glial cells in vivo. To further analyze these interactions, the patch-clamp technique was combined with single-cell RT-PCR in acute hippocampal brain slices. Subsequent to functional analysis, the cytoplasm of the same cell was harvested to perform transcript analysis and identify subunits that underlie inwardly rectifying K+ currents (I(Kir)) in astrocytes of the CA1 stratum radiatum. Transcripts encoding Kir2.1, Kir2.2, or Kir2.3, were encountered in a majority of cells, while Kir4.1 was less frequent. Further investigation revealed that glial Kir channels are rapidly inhibited upon activation of AMPA-type glutamate receptors, most probably due a receptor mediated influx of Na+, which plugs the channels from the intracellular side. A transient inhibition of I(Kir) in astrocytes in response to neuronal glutamate release and glial AMPA receptor activation represents a further, so far undetected mechanism to balance neuronal excitability. PMID- 11906216 TI - The neuron-specific Ca2+-binding protein caldendrin: gene structure, splice isoforms, and expression in the rat central nervous system. AB - Caldendrin is the founder member of a recently discovered family of calmodulin like proteins, which are highly abundant in brain. In this study we examined the organization of the murine and human caldendrin gene as well as the expression pattern of transcripts for caldendrin and two novel splice variants. In addition the distribution of caldendrin in rat brain has been assessed by immunohistochemistry. Caldendrin is localized to the somatodendritic compartment of a subpopulation of mainly principal neurons in brain regions with a laminar organization and is present only at a subset of mature excitatory synapses. Caldendrin immunoreactivity (IR) is tightly associated with the cortical cytoskeleton, enriched in the postsynaptic density (PSD) fraction, and associates late during development with the synaptic cytomatrix. The expression is highly heterogenous within cortex, with highest levels of caldendrin IR in layer III of the piriform and layer II/III of the somatosensory cortex. The segregated cortical distribution to areas, which represent the most important primary sensory systems of the rodent brain, may reflect different requirements for dendritic Ca2+-signaling in these neurons. The presence of caldendrin in the PSD of distinct synapses may have important implications for Ca2+-modulated processes of synaptic plasticity. PMID- 11906217 TI - Phonetic perception and the temporal cortex. AB - Recent functional neuroimaging studies have emphasized the role of the different areas within the left superior temporal sulcus (STS) for the perception of various speech stimuli. We report here the results of three independent studies additionally demonstrating hemodynamic responses in the vicinity of the planum temporale (PT). In these studies we used consonant-vowel (CV) syllables, tones, white noise, and vowels as acoustic stimuli in the context of whole-head functional magnetic resonance imaging, applying a long TR to attenuate possible masking effects by the scanner noise. To summarize, we obtained the following results for the contrasts comparing hemodynamic responses obtained during the perception of CV syllables compared to tones or white noise: (i) stronger activation in the vicinity of the left PT with two distinct foci of activation, one in a lateral position and the other more medial in the vicinity of Heschl's sulcus; (ii) stronger activation in the vicinity of the right PT; and (iii) stronger bilateral activation within the mid-STS. Further contrasts revealed the following findings: (iv) stronger bilateral activation to CV syllables than to vowels in the medial PT, (v) stronger left-sided activation to CV syllables than to vowels in the mid-STS, and (vi) stronger activation to CV syllables with voiceless initial consonants than to CV syllables with voiced initial consonants in the left medial PT. The results are compatible with the hypothesis that the STS contains neurons specialized for speech perception. However, these results also emphasize the role of the PT in the analysis of phonetic features, namely the voice-onset-time. Yet this does not mean that the PT is solely specialized for phonetic analysis. We hypothesize rather that the PT contains neurons specialized for the analysis of rapidly changing cues as was suggested by P. Tallal et al. (1993, Ann. N. Y. Acad. Sci. 682: 27-47). PMID- 11906218 TI - The quantitative evaluation of functional neuroimaging experiments: the NPAIRS data analysis framework. AB - We introduce a data-analysis framework and performance metrics for evaluating and optimizing the interaction between activation tasks, experimental designs, and the methodological choices and tools for data acquisition, preprocessing, data analysis, and extraction of statistical parametric maps (SPMs). Our NPAIRS (nonparametric prediction, activation, influence, and reproducibility resampling) framework provides an alternative to simulations and ROC curves by using real PET and fMRI data sets to examine the relationship between prediction accuracy and the signal-to-noise ratios (SNRs) associated with reproducible SPMs. Using cross validation resampling we plot training-test set predictions of the experimental design variables (e.g., brain-state labels) versus reproducibility SNR metrics for the associated SPMs. We demonstrate the utility of this framework across the wide range of performance metrics obtained from [(15)O]water PET studies of 12 age- and sex-matched data sets performing different motor tasks (8 subjects/set). For the 12 data sets we apply NPAIRS with both univariate and multivariate data analysis approaches to: (1) demonstrate that this framework may be used to obtain reproducible SPMs from any data-analysis approach on a common Z-score scale (rSPM[Z]); (2) demonstrate that the histogram of a rSPM[Z] image may be modeled as the sum of a data-analysis-dependent noise distribution and a task-dependent, Gaussian signal distribution that scales monotonically with our reproducibility performance metric; (3) explore the relation between prediction and reproducibility performance metrics with an emphasis on bias-variance tradeoffs for flexible, multivariate models; and (4) measure the broad range of reproducibility SNRs and the significant influence of individual subjects. A companion paper describes learning curves for four of these 12 data sets, which describe an alternative mutual-information prediction metric and NPAIRS reproducibility as a function of training-set sizes from 2 to 18 subjects. We propose the NPAIRS framework as a validation tool for testing and optimizing methodological choices and tools in functional neuroimaging. PMID- 11906219 TI - The quantitative evaluation of functional neuroimaging experiments: mutual information learning curves. AB - Learning curves are presented as an unbiased means for evaluating the performance of models for neuroimaging data analysis. The learning curve measures the predictive performance in terms of the generalization or prediction error as a function of the number of independent examples (e.g., subjects) used to determine the parameters in the model. Cross-validation resampling is used to obtain unbiased estimates of a generic multivariate Gaussian classifier, for training set sizes from 2 to 16 subjects. We apply the framework to four different activation experiments, in this case [(15)O]water data sets, although the framework is equally valid for multisubject fMRI studies. We demonstrate how the prediction error can be expressed as the mutual information between the scan and the scan label, measured in units of bits. The mutual information learning curve can be used to evaluate the impact of different methodological choices, e.g., classification label schemes, preprocessing choices. Another application for the learning curve is to examine the model performance using bias/variance considerations enabling the researcher to determine if the model performance is limited by statistical bias or variance. We furthermore present the sensitivity map as a general method for extracting activation maps from statistical models within the probabilistic framework and illustrate relationships between mutual information and pattern reproducibility as derived in the NPAIRS framework described in a companion paper. PMID- 11906220 TI - Predicting perceptual events activates corresponding motor schemes in lateral premotor cortex: an fMRI study. AB - The ability to recognize sequential patterns of external events enables us to predict their future course and thus to plan and execute actions based on current perceptions and previous experiences. Here we show with functional magnetic resonance imaging that even in the absence of movement the prediction of sequential patterns activates brain areas involved in the representation of specific motor schemas. Particularly, the prediction of size engages premotor areas involved in hand movements (superior part of the ventrolateral premotor cortex), whereas the prediction of pitch engages premotor areas involved in articulation (inferior most ventrolateral premotor cortex). The findings indicate that events are mapped onto somatotopically corresponding motor schemes whenever we predict sequential perceptions. PMID- 11906221 TI - Initial demonstration of in vivo tracing of axonal projections in the macaque brain and comparison with the human brain using diffusion tensor imaging and fast marching tractography. AB - Diffusion tensor imaging (DTI), a magnetic resonance imaging technique, is used to infer major axonal projections in the macaque and human brain. This study investigates the feasibility of using known macaque anatomical connectivity as a "gold-standard" for the evaluation of DTI tractography methods. Connectivity information is determined from the DTI data using fast marching tractography (FMT), a novel tract-tracing (tractography) method. We show for the first time that it is possible to determine, in an entirely noninvasive manner, anatomical connection pathways and maps of an anatomical connectivity metric in the macaque brain using a standard clinical scanner and that these pathways are consistent with known anatomy. Analogous human anatomical connectivity is also presented for the first time using the FMT method, and the results are compared. The current limitations of the methodology and possibilities available for further studies are discussed. PMID- 11906222 TI - The synchronization of the human cortical working memory network. AB - A verbal reasoning problem at the intersection of verbal working memory, problem solving, and language comprehension was examined using event-related fMRI to distinguish differences in the differential timing of the response of the various cortical regions that compose the working memory network. Problems were developed such that the process demand as well as the timing of the manipulation of the contents of working memory (i.e., a demanding computation) was varied. Activation was observed in several regions including the dorsolateral prefrontal cortex, the inferior frontal gyrus, and the parietal lobe. Examination of the MR amplitude response revealed that the regions do not all activate simultaneously; instead, their activation time courses reveal differential responses that correspond to their theoretical processing role in the problem-solving task. The coordination of cortical area responses reveals how the various cortical regions synchronize and collaborate in order to accomplish a given cognitive function. PMID- 11906223 TI - Single-trial variability in event-related BOLD signals. AB - Most current analysis methods for fMRI data assume a priori knowledge of the time course of the hemodynamic response (HR) to experimental stimuli or events in brain areas of interest. In addition, they typically assume homogeneity of both the HR and the non-HR "noise" signals, both across brain regions and across similar experimental events. When HRs vary unpredictably, from area to area or from trial to trial, an alternative approach is needed. Here, we use Infomax independent component analysis (ICA) to detect and visualize variations in single trial HRs in event-related fMRI data. Six subjects participated in four fMRI sessions each in which ten bursts of 8-Hz flickering-checkerboard stimulation were presented for 0.5-s (short) or 3-s (long) durations at 30-s intervals. Five axial slices were acquired by a Bruker 3-T magnetic resonance imager at interscan intervals of 500 ms (TR). ICA decomposition of the resulting blood oxygenation level-dependent (BOLD) data from each session produced an independent component active in primary visual cortex (V1) and, in several sessions, another active in medial temporal cortex (MT/V5). Visualizing sets of BOLD response epochs with novel BOLD-image plots demonstrated that component HRs varied substantially and often systematically across trials as well as across sessions, subjects, and brain areas. Contrary to expectation, in four of the six subjects the V1 component HR contained two positive peaks in response to short-stimulus bursts, while components with nearly identical regions of activity in long-stimulus sessions from the same subjects were associated with single-peaked HRs. Thus, ICA combined with BOLD-image visualization can reveal dramatic and unforeseen HR variations not apparent to researchers analyzing their data with event-related response averaging and fixed HR templates. PMID- 11906225 TI - Anatomical variability of the anterior cingulate gyrus and basic dimensions of human personality. AB - This study focused on investigating a possible relationship between interindividual variability in the morphology of the cingulate gyrus and behavioral styles. Using magnetic resonance images obtained from 100 healthy young volunteers (50 women and 50 men), we measured the surface area of the anterior cingulate gyrus and related it to the scores on the Temperament and Character Inventory. Anatomical data revealed that hemispheric asymmetry in the anterior cingulate gyrus surface area was very common (83% of cases) and that a prominent right anterior cingulate was more frequent in women than in men. In the correlational analysis, surface measurements of the right anterior cingulate gyrus accounted for a 24% score variance in Harm Avoidance. Both women and men with larger right anterior cingulate described themselves as experiencing greater worry about possible problems, fearfulness in the face of uncertainty, shyness with strangers, and fatigability. Furthermore, women reported overall higher scores in Harm Avoidance than men; these gender differences were largely explained by gender differences in the right anterior cingulate area in a covariate analysis. Our observations suggest that a large right anterior cingulate is related to a temperamental disposition to fear and anticipatory worry in both genders and that a higher prevalence of these traits in women may be coupled with a greater expansion of this brain region. PMID- 11906224 TI - Error rate and outcome predictability affect neural activation in prefrontal cortex and anterior cingulate during decision-making. AB - Decision-making in the presence of uncertainty is a complex process that involves both affective and cognitive factors. Both error rate and predictability have been implicated in the process of response selection during decision-making. This study examined the hypothesis that the rate of errors during decision-making differentially affects the activation in prefrontal and cingulate cortex. BOLD echo-planar signal intensity was obtained during a two-choice prediction task across 90-s blocks with 20, 50, or 80% error rates. This study yielded three main findings. First, at chance level error rates, activation of the right dorsolateral (BA 9, 46), inferior prefrontal (BA 44), and precuneus (BA 7) during the two-choice prediction task replicated the finding previously reported. Second, premotor (BA 6) and parahippocampal (BA 36) areas were relatively more active at high error rates, and dorsolateral (BA 9, 46) and inferior prefrontal cortex (BA 44) as well as parietal (BA 40) and cingulate cortex (BA 25, 32) were more active during low error rates. Third, the relationship between the frequency of the dominant strategy underlying decision-making (win-stay/lose-shift) and the activation in the dorsolateral prefrontal cortex and the anterior cingulate was dependent on error rate or outcome predictability. These results support the hypothesis that error rates and predictability affect the activation patterns in the neural systems underlying decision-making because these structures maintain a representation of the reinforcement history for the available response alternatives to select an "optimal strategy." PMID- 11906226 TI - Dynamic brain activation during processing of emotional intonation: influence of acoustic parameters, emotional valence, and sex. AB - Appreciation of the emotional tone of verbal utterances represents an important aspect of social life. It is still unsettled, however, which brain areas mediate processing of intonational information and whether the presumed right-sided superiority depends upon acoustic properties of the speech signal. Functional magnetic resonance imaging was used to disentangle brain activation associated with (i) extraction of specific acoustic cues and (ii) detection of specific emotional states. Stimulus material comprised pairs of emotionally intonated utterances, exclusively differing either in pitch range or in the length of stressed vowels. Hemodynamic responses showed a dynamic pattern of cerebral activation including sequenced bilateral responses of various cortical and subcortical structures. Activation associated with discrimination of emotional expressiveness predominantly emerged within the right inferior parietal lobule, within the bilateral mesiofrontal cortex and--with an asymmetry toward the right hemisphere--at the level of bilateral dorsolateral frontal cortex. Lateralization did not depend upon acoustic structure or emotional valence of stimuli. These findings might prove helpful in reconciling the controversial previous clinical and experimental data. PMID- 11906227 TI - Thresholding of statistical maps in functional neuroimaging using the false discovery rate. AB - Finding objective and effective thresholds for voxelwise statistics derived from neuroimaging data has been a long-standing problem. With at least one test performed for every voxel in an image, some correction of the thresholds is needed to control the error rates, but standard procedures for multiple hypothesis testing (e.g., Bonferroni) tend to not be sensitive enough to be useful in this context. This paper introduces to the neuroscience literature statistical procedures for controlling the false discovery rate (FDR). Recent theoretical work in statistics suggests that FDR-controlling procedures will be effective for the analysis of neuroimaging data. These procedures operate simultaneously on all voxelwise test statistics to determine which tests should be considered statistically significant. The innovation of the procedures is that they control the expected proportion of the rejected hypotheses that are falsely rejected. We demonstrate this approach using both simulations and functional magnetic resonance imaging data from two simple experiments. PMID- 11906228 TI - Dissociation of working memory processing associated with native and second languages: PET investigation. AB - Verbal working memory plays a significant role in language comprehension and problem-solving. The prefrontal cortex has been suggested as a critical area in working memory. Given that domain-specific dissociations of working memory may exist within the prefrontal cortex, it is possible that there may also be further functional divisions within the verbal working memory processing. While differences in the areas of the brain engaged in native and second languages have been demonstrated, little is known about the dissociation of verbal working memory associated with native and second languages. We have used H2(15)O positron emission tomography in 14 normal subjects in order to identify the neural correlates selectively involved in working memory of native (Korean) and second (English) languages. All subjects were highly proficient in the native language but poorly proficient in the second language. Cognitive tasks were a two-back task for three kinds of visually presented objects: simple pictures, English words, and Korean words. The anterior portion of the right dorsolateral prefrontal cortex and the left superior temporal gyrus were activated in working memory for the native language, whereas the posterior portion of the right dorsolateral prefrontal cortex and the left inferior temporal gyrus were activated in working memory for the second language. The results suggest that the right dorsolateral prefrontal cortex and left temporal lobe may be organized into two discrete, language-related functional systems. Internal phonological processing seems to play a predominant role in working memory processing for the native language with a high proficiency, whereas visual higher order control does so for the second language with a low proficiency. PMID- 11906229 TI - Three-dimensional diffusion tensor magnetic resonance microimaging of adult mouse brain and hippocampus. AB - The use of diffusion tensor information as an additional contrast in MR microimaging was investigated in ex vivo mouse brain and isolated hippocampus. Diffusion Tensor Imaging (DTI) provided unique contrast to identify many internal structures of the gray matter such as hippocampus, thalamus, and cortex. In hippocampus, stratum granulosum and stratum pyramidale could be identified using the isotropic water diffusion constant. Stratum moleculare and stratum radiatum were identified from their characteristic fiber architecture revealed by color coded DTI. Identification of these structures allowed reconstruction of their 3-D volume. Thus, high-resolution DTI has excellent potential as a tool for 3-D characterization of murine brains. PMID- 11906230 TI - Comparison of detrending methods for optimal fMRI preprocessing. AB - Because of the inherently low signal to noise ratio (SNR) of fMRI data, removal of low frequency signal intensity drift is an important preprocessing step, particularly in those brain regions that weakly activate. Two known sources of drift are noise from the MR scanner and aliasing of physiological pulsations. However, the amount and direction of drift is difficult to predict, even between neighboring voxels. Further, there is no concensus on an optimal baseline drift removal algorithm. In this paper, five voxel-based detrending techniques were compared to each other and an auto-detrending algorithm, which automatically selected the optimal method for a given voxel time-series. For a significance level of P < 10(-6), linear and quadratic detrending moderately increased the percentage of activated voxels. Cubic detrending decreased activation, while a wavelet approach increased or decreased activation, depending on the dataset. Spline detrending was the best single algorithm. However, auto-detrending (selecting the best algorithm or none, if detrending is not useful) appears to be the most judicious choice, particularly for analyzing fMRI data with weak activations in the presence of baseline drift. PMID- 11906231 TI - Neural correlates of spontaneous direction reversals in ambiguous apparent visual motion. AB - Looking at bistable visual stimuli, the observer experiences striking transitions between two competing percepts while the physical stimulus remains the same. Using functional imaging techniques, it is therefore possible to isolate neural correlates of perceptual changes that are independent of the low-level aspects of the stimulus. Previous experiments have demonstrated distributed activations in human extrastriate visual cortex related to switches between competing percepts. Here we asked where extrastriate responses still occur with a bistable stimulus that minimizes the cognitive difference between the two percepts. We used the "spinning wheel illusion," a bistable apparent motion stimulus of which both possible percepts correspond to the same object, share the same center, and are perceived as identically patterned stimuli moving at the same speed and changing only in direction. Using functional magnetic resonance imaging, we analyzed the spatial distribution of event-related activations occurring during spontaneous reversals of perceived direction of motion. In accordance with earlier neuroimaging findings for bistable percepts, we observed event-related activations in several frontal and parietal areas, including the superior parietal cortex bilaterally, the right inferior parietal cortex, and the premotor and inferior frontal cortex of both hemispheres. Furthermore, we found bilateral activations in the occipitotemporal junction (hMT+/V5) and in the lateral occipital sulcus ("KO") posterior to hMT+/V5, but not in areas of the "ventral stream" of cortical visual processing. Our data suggest that, while a frontoparietal network subserves more general aspects in bistable visual perception, the activations in functionally specialized extrastriate visual cortex are highly category- or attribute-specific. PMID- 11906232 TI - Retrieval of visual, auditory, and abstract semantics. AB - Conceptual knowledge is thought to be represented in a large distributed network, indexing a range of different semantic features (e.g., visual, auditory, functional). We investigated the anatomical organization of these features, using PET, by contrasting brain activity elicited by heard words with (i) visual (e.g., blue), (ii) auditory (e.g., noise), or (iii) abstract (e.g., truth) meaning. The activation task was either repetition or semantic decision (e.g., does the meaning of the word relate to religion?). In the baseline conditions, the sound track of the words was reversed and subjects had to say "OK" (control for repetition) or make an acoustic decision (control for semantic decision). Irrespective of task, words relative to their corresponding controls activated the left posterior inferior temporal and inferior frontal cortices. In addition, semantic decisions on words with sensory (visual and auditory) meanings enhanced activation in a ventral region of the left anterior temporal pole. These results are consistent with neuropsychological studies showing that anterior temporal lobe damage can cause deficits for items that are mainly defined by their sensory features (i.e., concrete, particularly living items). Since modality-specific activation was observed only during the semantic decision task, we discuss whether it reflects retrieval of sensory semantics per se or the degree to which semantic associations are triggered during effortful retrieval. PMID- 11906233 TI - A PET study of stimulus- and task-induced semantic processing. AB - To investigate the neural correlates of semantic processing, previous functional imaging studies have used semantic decision and generation tasks. However, in addition to activating semantic associations these tasks also involve executive functions that are not specific to semantics. The study reported in this paper aims to dissociate brain activity due to stimulus-driven semantic associations and task-induced semantic and executive processing by using repetition and semantic decision on auditorily presented words in a cognitive conjunction design. The left posterior inferior temporal, inferior frontal (BA 44/45), and medial orbital gyri were activated by both tasks, suggesting a general role in stimulus-driven semantic and phonological processing. In addition, semantic decision increased activation in (i) left ventral inferior frontal cortex (BA 47), right cerebellum, and paracingulate, which have all previously been implicated in executive functions, and (ii) a ventral region in the left anterior temporal pole which is commonly affected in patients with semantic impairments. We attribute activation in this area to the effortful linkage of semantic features. Thus, our study replicated the functional dissociation between dorsal and ventral regions of the left inferior frontal cortex. Moreover, it also dissociated the semantic functions of the left posterior inferior temporal gyrus and anterior temporal pole: The posterior region subserves stimulus-driven activation of semantic associations and the left anterior region is involved in task-induced association of semantic information. PMID- 11906234 TI - The neural basis for category-specific knowledge: an fMRI study. AB - Functional neuroimaging studies of healthy adults have associated different categories of knowledge with distinct activation patterns. The basis for these recruitment patterns has been controversial, due in part to the limited range of categories that has been studied. We used fMRI to monitor regional cortical recruitment patterns while subjects were exposed to printed names of Animals, Implements, and Abstract nouns. Both Implements and Abstract nouns were related to recruitment of left posterolateral temporal cortex and left prefrontal cortex, and Abstract nouns additionally recruited posterolateral temporal and prefrontal regions of the right hemisphere. Animals were associated with activation of ventral-medial occipital cortex in the left hemisphere at a level that approaches significance. These findings are not consistent with the "sensory-motor" model proposed to explain the neural representation of word knowledge. We suggest instead a neural model of semantic memory that reflects the processes common to understanding Implements and Abstract nouns and a selective sensitivity, possibly evolving from adaptive pressures, to the overlapping, intercorrelated visual characteristics of Animals. PMID- 11906235 TI - Increased FDG uptake in the ipsilesional sensorimotor cortex in congenital hemiplegia. AB - The resting brain metabolism was estimated in six children suffering from a right congenital hemiplegia (CH) of subcortical origin. This estimate was based on the 18F-labeled 2-deoxy-2-fluoro-d-glucose (FDG) uptake measured by means of positron emission tomography and compared, using statistical parametric mapping (SPM99), with that of six control subjects. The contrast [CH children - Controls] showed that CH children had two loci of relatively higher FDG uptake. The larger voxel cluster was found in the ipsilesional hemisphere and comprised the primary motor and somatosensory cortices and left inferior parietal lobule. The other cluster was located in the contralesional hemisphere and encompassed the primary motor cortex, callosomarginal sulcus, and cingulate gyrus. The reverse contrast [Controls - CH children] showed that control subjects had a relatively higher FDG uptake bilaterally in the temporal and hippocampal gyri, the rostral part of the brain stem, the thalami, the putamen, and the superior frontal gyri. A crossed cerebellar diaschisis was not observed in CH children. This relatively higher FDG uptake in the ipsi- and contralesional motor areas of CH children stands out in contrast to the hypometabolism (diaschisis) frequently observed in adult stroke patients with a subcortical lesion. This increased FDG uptake in the disconnected ipsilesional motor areas may reflect a long-term adaptation leading, for example, to an increased synaptic density and/or activity or to a change in the density of glucose transporters. PMID- 11906236 TI - Activation of attention networks using frequency analysis of a simple auditory motor paradigm. AB - The purpose of this study was to devise a paradigm that stimulates attention using a frequency-based analysis of the data acquired during a motor task. Six adults (30-40 years of age) and one child (10 years) were studied. Each subject was requested to attend to "start" and "stop" commands every 20 s alternatively and had to respond with the motor task every second time. Attention was stimulated during a block-designed, motor paradigm in which a start-stop commands cycle produced activation at the fourth harmonic of the motor frequency. We disentangled the motor and attention functions using statistical analysis with subspaces spanned by vectors generated by a truncated trigonometric series of motor and attention frequency. During our auditory-motor paradigm, all subjects showed activation in areas that belong to an extensive attention network. Attention and motor functions were coactivated but with different frequencies. While the motor-task-related areas were activated with slower frequency than attention, the activation in the attention-related areas was enhanced every time the subject had to start or end the motor task. We suggest that although a simple block-designed, auditory-motor paradigm stimulates the attention network, motor preparation, and motor inhibition concurrently, a frequency-based analysis can distinguish attention from motor functions. Due to its simplicity the paradigm can be valuable in studying children with attention deficit disorders. PMID- 11906237 TI - Functional anatomy of visual search: regional segregations within the frontal eye fields and effective connectivity of the superior colliculus. AB - The ability to find targets embedded within complex visual environments requires the dynamic programming of visuomotor search behaviors. Functional magnetic resonance imaging was used to image subjects while they visually searched for targets embedded among foils. Visuomotor search activated the posterior parietal cortex and the frontal eye fields. Both regions showed a greater number of activated voxels on the right, consistent with the known pattern of right hemispheric dominance for spatial attention. The superior colliculus showed prominent activation in the search versus eye movement contrast, demonstrating, for the first time in humans, activation of this region specifically related to an exploratory attentional contingency. An analysis of effective connectivity demonstrated that the search-dependent variance in the activity of the superior colliculus was significantly influenced by the activity in a network of cortical regions including the right frontal eye fields and bilateral parietal and occipital cortices. These experiments also revealed the presence of a mosaic of activated sites within the frontal eye field region wherein saccadic eye movements, covert shifts of attention, and visuomotor search elicited overlapping but not identical zones of activation. In contrast to the existing literature on functional imaging, which has focused on covert shifts of spatial attention, this study helps to characterize the functional anatomy of overt spatial exploration. PMID- 11906238 TI - The anterior frontomedian cortex and evaluative judgment: an fMRI study. AB - This study investigated the neuronal basis of evaluative judgment. Judgments can be defined as the assessment of an external or internal stimulus on an internal scale and they are fundamental for decision-making and other cognitive processes. Evaluative judgments (I like George W. Bush: yes/no) are a special type of judgment, in which the internal scale is related to the person's value system (preferences, norms, aesthetic values, etc.). We used functional magnetic resonance imaging to examine brain activation during the performance of evaluative judgments as opposed to episodic and semantic memory retrieval. Evaluative judgment produced significant activation in the anterior frontomedian cortex (BA 10/9), the inferior precuneus (BA 23/31), and the left inferior prefrontal cortex (BA 45/47). The results show a functional dissociation between the activations in the anterior frontomedian cortex and in the inferior precuneus. The latter was mainly activated by episodic retrieval processes, supporting its function as a multimodal association area that integrates the different aspects of retrieved and newly presented information. In contrast, the anterior frontomedian cortex was mainly involved in evaluative judgments, supporting its role in self-referential processes and in the self-initiation of cognitive processes. PMID- 11906239 TI - Rapid simultaneous mapping of T2 and T2* by multiple acquisition of spin and gradient echoes using interleaved echo planar imaging (MASAGE-IEPI). AB - A new MRI sequence for the rapid simultaneous measurement of T2 and T2* is presented. The technique uses the multiple acquisition of spin and gradient echoes with interleaved echo planar imaging (MASAGE-IEPI). IEPI data sets are sampled during and between a pair of short and long echo time spin echoes, allowing the reconstruction of a set of images with different combinations of T2 and T2* weighting and the calculation of T2 and T2* maps. In the context of neuroimaging, these maps can provide information on cerebral hemodynamics and oxygenation status, either via the deoxyhemoglobin-based BOLD signal or by the effect of exogenous paramagnetic contrast agents. MASAGE-IEPI benefits from the inherent advantages of the IEPI approach, i.e., high time resolution and minimal image distortion, and also has good time efficiency due to the acquisition of multiple image data sets following each excitation pulse. The accuracy of the sequence for the measurement of T2 and T2* is verified on phantoms, and the technique is applied to monitor changing hemodynamics in the rat brain during episodes of hypoxia. Data for the generation of maps of T2 and T2* are acquired with a time resolution of 12 s to accurately define the rapidly changing time course. As increasing emphasis is placed on the role of T2 and T2* in the direct measurement of physiological parameters such as cerebral metabolic rate of oxygen consumption and blood vessel sizes, MASAGE-IEPI offers an efficient method for the measurement of these two important MRI parameters. PMID- 11906240 TI - Brain activation modulated by the comprehension of normal and pseudo-word sentences of different processing demands: a functional magnetic resonance imaging study. AB - Recent data from lesion and brain imaging studies have questioned the well established assumption of a close functional-anatomic link between syntax and Broca's area and semantics and Wernicke's area. In the present study we used functional magnetic resonance imaging (fMRI) to investigate the neuroanatomical correlates of semantic and syntactic functions and possible interdependencies between the related brain systems. In a completely crossed design we varied syntactic processing demands (easy vs difficult to process word order sequences) and the meaningfulness of sentences (real- vs pseudo-word sentences). In comparison to a backward speech condition we found an activation of the left perisylvian region, including the left inferior frontal cortex and the left superior and middle temporal gyri. Semantic in contrast to pseudo-word sentences elicited a stronger activation in both the anterior and the posterior perisylvian cortex. Syntactic difficulty had its strongest effect within the left inferior frontal region and this effect was more pronounced for semantic than nonsemantic speech. These results suggest that semantic and syntactic language functions are mediated by partly specialized brain systems but that there nevertheless exists a substantial functional overlap of the involved brain structures. PMID- 11906241 TI - Multiparametric quantitation of the perilesional region in patients with healed or healing solitary cysticercus granuloma. AB - The purpose of this study was to compute T2 values and magnetization transfer (MT) ratios in the perilesional region of healing and healed cysticercus granulomas to determine if there are T2 abnormalities not apparent on conventional T2-weighted imaging and to determine the relationship between seizure control and the quantitative measures. Sixty-three patients were studied. T2 values and MT ratios were computed for the perilesional region and were compared with measurements from the contralateral normal-appearing region. A significantly increased T2 value was found for the perilesional region compared to the corresponding contralateral region despite the absence of qualitative abnormality on conventional T2-weighted magnetic resonance imaging. For patients showing normal-appearing perilesional regions on MT imaging, there was no significant difference in T2 and MT ratios between the perilesional and the normal contralateral regions. There was a statistically significant inverse correlation between perilesional T2 values and MT ratios, suggesting each was associated with perilesional gliosis. The study illustrates that quantitative evaluation of MT ratios and T2 augments the qualitative visual assessment of the perilesional region in healing or healed cysticercus granulomas. PMID- 11906242 TI - Cleavage of the fifth component of human complement and release of a split product with C5a-like activity by crystalline silica through free radical generation and kallikrein activation. AB - The effects of the same form of crystalline silica variously modified were compared to investigate the mechanisms by which silica activates C5 molecules. After incubation in human plasma, silica generated C5a-type fragments that stimulated polymorphonuclear leukocyte chemotaxis. This activity was totally abolished when plasma, adsorbed with antiserum against C5a or thermally inactivated, was used. Pretreatment of plasma with deferoxamine, 1,3 dimethyl-2 thiourea, or aprotinin markedly inhibited or totally abolished C5 activation. Finally, a significant increase in kallikrein activity was detected after incubation of silica particles in plasma. The results seem to indicate that the activation of C5 by crystalline silica occurs through a complex mechanism: the redox-active iron possibly present at the silica surface catalyzes, via Haber Weiss cycles, the production of hydroxyl radicals, which in turn convert native C5 to an oxidized C5-like form. This product is then cleaved by kallikrein, activated by the same silica particles, yielding oxidized C5a with the same functional properties as C5a. The different types of the same form of silica exhibited different reactivity. Two separate properties of the dusts seem to contribute to C5 activation: the potential to release hydroxyl radicals and the extent of C5 adsorption at the surface. The degree of surface hydrophobicity/hydrophilicity appeared sufficient to explain the different responses. PMID- 11906243 TI - The polychlorinated biphenyl mixture aroclor 1254 induces death of rat cerebellar granule cells: the involvement of the N-methyl-D-aspartate receptor and reactive oxygen species. AB - Polychlorinated biphenyls (PCBs) are widespread persistent environmental contaminants that display a complex spectrum of toxicological properties, including neurotoxicity. The present study investigates the effects of the PCB mixtures Aroclor 1242 (A1242) and Aroclor 1254 (A1254), and the PCB congeners 126 (3,3',4,4',5,-PeCB) and 153 (2,2',4,4',5,5'-HxCB) on formation of reactive oxygen species (ROS) and cell death in cultured rat cerebellar granule cells. The increase of ROS and induction of cell death were assayed using the fluorescent probe 2,7-dichlorofluorescin diacetate (DCFH-DA) and the trypan blue exclusion assay, respectively. A1242 and A1254 and PCB 153 induced a concentration dependent increase in cell death and ROS formation. A1254 was selected for mechanistic studies. When the cerebellar granule cells were exposed to 15 microM A1254 for 12 h, 95% of the cells died. Both PCB-mediated cell death and the increase of the ROS formation were inhibited by MK-801, demonstrating the importance of the N-methyl-D-aspartate receptor. Inhibitors of nitric oxide synthase and phospholipase A2 led to a significant reduction of the DCF fluorescence and cell death. The mitochondrial permeability transition pore blocker cyclosporin A and the antioxidant vitamin E also increased survival and reduced ROS formation. The results show a connection between cell death and free radical formation. PMID- 11906244 TI - Cytokine profiling for chemical sensitizers: application of the ribonuclease protection assay and effect of dose. AB - Exposure to chemicals in domestic and occupational settings may contribute to increases in asthma and allergy. Airway hypersensitivity (AHS) is T helper-2 (Th2) cell associated, whereas contact hypersensitivity (CHS) is T helper-1 (Th1) cell associated. The distinct cytokine profiles produced by these cells may provide a means of distinguishing respiratory sensitizers from contact sensitizers. In this study, female BALB/c mice were exposed twice on the flanks and three times on the ears using the airway sensitizer trimellitic anhydride (TMA) or the contact sensitizer dinitrochlorobenzene (DNCB). At various times following exposure, total mRNA was extracted from draining lymph node cells and cytokine mRNA profiles analyzed using a multiprobe ribonuclease protection assay (RPA). The Th2 cytokines IL4, IL10, and IL13 were significantly increased in response to TMA compared to DNCB, with optimal detection occurring 14 days following initial exposure. To determine its effect, dose was varied in flank exposures, ear exposures, or both simultaneously. When dose was varied during flank exposures only, TMA induced higher levels of Th2 cytokines than DNCB at all doses tested. DNCB did not induce Th1 cytokines at any dose tested. Variation of TMA dose during both exposures similarly induced Th2 cytokines. Dose only appeared to be a factor when TMA concentration was varied during the ear exposures alone. Thus, these studies suggest that quantitative differences in Th2 responses between TMA and DNCB may be demonstrated over a wide range of doses and these differences may be detected by RPA following dermal exposure to these sensitizers. PMID- 11906245 TI - Tumor necrosis factor-alpha-null mice are not resistant to cadmium chloride induced hepatotoxicity. AB - Acute administration of cadmium results in hepatotoxicity. Recent reports indicate that Kupffer cells, the resident macrophages of the liver, participate in the manifestation of chemical-induced hepatotoxicity. Tumor necrosis factor alpha (TNF-alpha) is a proinflammatory cytokine that is a major product of Kupffer cells and mediates the hepatotoxic effects of lipopolysaccharide (LPS). It has been speculated that cadmium also may exert its hepatotoxicity via the production of TNF-alpha by the Kupffer cells. Therefore, this study was undertaken to determine whether mice deficient in TNF-alpha are resistant to Cd induced hepatotoxicity. TNF-alpha-null (TNF-KO) and wild-type (WT) mice were dosed ip with saline, LPS (0.1 mg/kg)/Gln (d-galactosamine, 700 mg/kg), or CdCl2 (2.2, 2.8, 3.4, and 3.9 mg Cd/kg). Serum alanine aminotransferase (ALT) and sorbitol dehydrogenase (SDH) activities were quantified to assess liver injury. Caspase-3 activity was quantified to assess hepatocellular apoptosis. LPS/Gln treatment increased ALT (17-fold) and SDH (21-fold) in WT mice. In contrast, LPS/Gln-treatment did not significantly increase ALT or SDH in TNF-KO mice. LPS/Gln-treatment caused a 7.8-fold increase in caspase-3 activity in WT mice but did not increase caspase-3 in TNF-KO mice. Cadmium caused a dose-dependent increase in liver injury in both WT and TNF-KO mice. However, the liver injury produced by Cd in the TNF-KO mice was not different from that in WT at any dose. No significant increase in caspase-3 activity was detected in any of the Cd treated mice. These data indicate that, in contrast to LPS/Gln-induced hepatotoxicity, TNF-alpha does not appear to mediate Cd-induced hepatotoxicity. PMID- 11906246 TI - Renal toxicity of perchloroethylene and S-(1,2,2-trichlorovinyl)glutathione in rats and mice: sex- and species-dependent differences. AB - Suspensions of renal cells from rats and renal mitochondria from rats and mice were used to assess the sex and species dependence of acute toxicity due to perchloroethylene (Perc) and its glutathione conjugate S-(1,2,2 trichlorovinyl)glutathione (TCVG). A marked sex dependence in the acute cytotoxicity of both Perc and TCVG was observed: Perc caused significant release of lactate dehydrogenase (LDH) in isolated kidney cells from male but not female rats, and TCVG caused much more LDH release from male than female rat kidney cells. Assessment of toxicity in suspensions of isolated mitochondria from kidneys of male and female rats revealed a generally similar pattern of sensitivity, with mitochondria from males exhibiting significantly more inhibition of State 3 respiration and decrease of respiratory control ratio than mitochondria from females. Respiratory function in mitochondria from male and female mice, however, was also significantly inhibited by Perc or TCVG but exhibited little sex dependence in the degree of inhibition. Comparison with results from similar studies using the congener trichloroethylene and its glutathione conjugate suggested that Perc and TCVG are more potent nephrotoxicants. Neither Perc nor TCVG produced any significant effects on cytotoxicity or mitochondrial function in isolated hepatocytes from rats or in isolated liver mitochondria from rats or mice, suggesting that the liver is not a major acute target for Perc or its glutathione conjugate. Thus, many of the species-, sex-, and tissue-dependent differences in toxicity of Perc and TCVG that are observed in vivo are also observed in these in vitro models. PMID- 11906247 TI - Regulation of cell proliferation, apoptosis, and transcription factor activities during the promotion of liver carcinogenesis by polychlorinated biphenyls. AB - Polychlorinated biphenyls (PCBs) are environmental pollutants that are complete carcinogens and tumor promoters in the liver. The mechanisms of their promoting activities are not clear, but one possible mechanism is the induction of oxidative stress. In the present study we evaluated the ability of two PCB congeners to activate the oxidative stress-responsive transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1), as well as hepatocyte cell proliferation and apoptosis, which are influenced by activation of these transcription factors, in rat liver. Two transcription factors not activated by oxidative stress, signal transducers and activators of transcription 3 and 5 (STAT3 and STAT5), were also examined. All the animals in this study received a single dose of diethylnitrosamine (150 mg/kg) followed by four biweekly injections of 3,3',4,4'-tetrachlorobiphenyl (PCB-77) or 2,2',4,4',5,5' hexachlorobiphenyl (PCB-153) (100 or 300 micromol/kg), or both PCBs (100 micromol/kg each). Ten days after the last PCB injection, all animals were euthanized; 3 days before euthanasia all animals were implanted with Alzet osmotic pumps containing 5-bromo-2'-deoxyuridine (BrdU). The number of placental glutathione S-transferase (PGST)-positive foci were increased in rats administered PCBs, with the highest increase seen in rats administered PCB-77. The number of foci in rats administered both PCBs was intermediate between the numbers seen with either PCB-77 or PCB-153, indicating that a synergistic effect did not occur. There was a significant increase in NF-kappaB and AP-1 binding activities in hepatic nuclear extracts from rats receiving the high dose of PCB 77 or PCB-153 and in rats receiving both PCBs. In contrast, the DNA binding activities of STAT3 and STAT5 were decreased in rats administered PCBs. Cell proliferation in both focal and nonfocal hepatocytes was increased by PCB-77 but was not affected by PCB-153. Apoptotic indexes, as quantified by the TUNEL method, were increased in both focal and nonfocal hepatocytes by PCB-77 but were decreased in focal hepatocytes by PCB-153. This study shows that both PCBs alone or in combination can increase the DNA binding activities of NF-kappaB and AP-1, whereas the DNA binding activities of STAT3 and STAT5 are decreased. The induction of altered hepatic foci appears to be related to compensatory cell proliferation in PCB-77-treated rats, whereas the inhibition of apoptosis appears to be important in PCB-153-treated rats. PMID- 11906248 TI - Polychlorinated biphenyls interfere with androgen-induced transcriptional activation and hormone binding. AB - Polychlorinated biphenyls (PCBs) are ubiquitous highly persistent manufactured chemicals known to bioaccumulate in the food chain. Exposure to PCBs has been implicated in a wide range of human health effects, including altering normal endocrine processes and reproductive function. However, very little is understood regarding the specific mechanisms by which PCBs may exert their effects in biological systems. We have examined the ability of PCBs to interfere with transcriptional activation of the androgen receptor (AR) and glucocorticoid receptor (GR) in an in vitro transcription-based reporter assay system. Four Aroclor PCB mixtures were found to antagonize AR-mediated transcription in the presence of the natural AR ligand dihydrotestosterone (DHT). The antagonistic activity of Aroclor mixtures increased in the following order: 1260 < 1242 < 1254 < 1248. These Aroclor mixtures had no discernible effect on GR activity. Aroclor 1254 in the absence of DHT exhibited weak agonistic responses in a dose-dependent manner with AR. Within a series of individual congeners, congeners 42, 128, and 138 are shown to antagonize AR activity. These congeners all share a common core chlorine substitution pattern. Ligand-binding studies demonstrate that endocrine activities of PCB mixtures and congeners on AR are likely due to direct and specific binding to AR ligand-binding domain. PMID- 11906250 TI - Total analysis and purification of cellular proteins binding to cisplatin-damaged DNA using submicron beads. AB - A high-performance affinity purification technique has been developed for cisplatin (CDDP)-damaged DNA binding proteins directly from crude nuclear extracts of HeLaS3 cell using novel submicron beads synthesized by copolymerization of styrene and glycidyl methacrylate (GMA). The beads dramatically decreased both nonspecific protein adsorption on solid surfaces and elution volume and simplified the handling procedure. Preparation of the beads for purification was carried out by immobilization of telomeric repeats, (TTAGGG)(n), on the surface after the reaction with CDDP. At least nine proteins clearly showed higher affinity to CDDP-DNA and were identified by amino acid sequence analysis including HMGB (high mobility group), hUBF (human upstream binding factor), and Ku autoantigen, which were previously reported to be components of CDDP-damaged DNA binding proteins. PMID- 11906252 TI - Electrochemical detection of gene expression in tumor samples: overexpression of Rak nuclear tyrosine kinase. AB - Absolute quantification of Rak nuclear tyrosine kinase mRNA in breast tissue samples was determined by competitive RT-PCR. The total RNA from the same samples was also chemically amplified through conventional RT-PCR, and the relative amounts of these amplified RT-PCR products were determined by adsorption onto an indium tin oxide (ITO) electrode followed by electrochemical detection. The electrochemical detection was performed using the inorganic metal complex Ru(bpy)(3)(2+) (bpy = 2,2' bipyridine) to catalyze the oxidation of the guanine residues of the immobilized RT-PCR products. Using the competitive RT-PCR values as standards, it was found that an optimized conventional RT-PCR coupled with electrochemical detection provides a simple method for measuring relative gene expression among a series of mRNA samples from breast tumors. The use of electrochemical detection potentially eliminates the need for gel electrophoresis and fluorescent or radioactive labels in detecting the target genes. PMID- 11906251 TI - Formation of a bioconjugate composed of hemin, smectite, and quaternary ammonium chloride that is soluble and active in hydrophobic media. AB - Hemin (Fe(3+)) was adsorbed onto synthetic smectite (clay mineral) intercalated with a quaternary alkenylammonium compound, dioleyldimethylammonium chloride (DOA), to form a hemin-smectite-DOA conjugate. The hemin-smectite-DOA conjugate was soluble in organic solvents such as benzene and toluene to form a transparent colloidal solution with a light yellow color. Its absorption spectrum in benzene showed two bands, 600 and 568 nm, in the visible region and a sharp Soret band at 400 nm with the molar extinction coefficient of 7.5 x 10(4) M(-1) cm(-1). The formation of the conjugate of smectite and DOA was confirmed by X-ray diffraction analysis: the basal spacing, d(001), of hemin-smectite-DOA conjugate was 19 A which is an expansion of the interlayer space by 5 A based upon the basal spacing of smectite of 14 A. Hemin-smectite-DOA conjugate catalyzed the peroxidase-like reaction in organic solvents using benzoyl peroxide as the hydrogen acceptor and leucocrystal violet as the hydrogen donor. The temperature-dependent peroxidase like activity of the conjugate was compared with peroxidase activity of horseradish peroxidase. The hemin-smectite-DOA conjugate exhibited higher activity as the temperature was increased from 30 to 70 degrees C, while horseradish peroxidase activity was reduced as the temperature was increased. PMID- 11906254 TI - Reversible surface thiol immobilization of carboxyl group containing haptens to a BIAcore biosensor chip enabling repeated usage of a single sensor surface. AB - We describe a reversible immobilization method for carboxyl group containing haptens that makes the repeated usage of a BIAcore biosensor chip possible. Haptens which are immobilized according to the surface thiol method can be removed completely from the sensor surface again by a reducing step. In the first part of our study, analogues of the herbicides 2,4-dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid were immobilized in succession to a biosensor surface of a BIAcore surface plasmon resonance instrument according to the thiol coupling method. Direct kinetic analysis of these ligands to a polyclonal anti 2,4-dichlorophenoxyacetic acid antibody were performed using these biosensor surfaces. In the second part of the study, different amounts of 2,4 dichlorophenoxyacetic acid were sequentially immobilized onto the same biosensor surface in order to generate a calibration plot for 2,4-dichlorophenoxyacetic acid. Using this plot, the quantitative detection of the herbicide down to a concentration of 0.1 microg/mL, the maximum admissible concentration of pesticides in drinking water, is possible. PMID- 11906253 TI - Protamine-fragment peptides fused to an SV40 nuclear localization signal deliver oligonucleotides that produce antisense effects in prostate and bladder carcinoma cells. AB - The development of antisense technology has focused on improving methods for oligonucleotide delivery into cells. In the present work, we describe a novel strategy for oligonucleotide delivery based on a bifunctional peptide composed of a C-terminal protamine-fragment that contains a DNA-binding domain and an N terminal nuclear localization signal sequence derived from the SV40 large-T antigen (The sequences of two of the peptides are R6WGR6-PKKKRKV [s-protamine NLS] and R4SR6FGR6VWR4-PKKKRKV [l-protamine-NLS]). We demonstrated, by intrinsic fluorescence quenching, that peptides of this class form complexes with oligodeoxynucleotides. To evaluate delivery, we used a 20-mer phosphorothioate oligomer (Isis 3521) targeted to the 3'-untranslated region of the PKC-alpha mRNA and G3139, an 18-mer phosphorothioate targeted to the first six codons of the human bcl-2 open reading frame, and complexed them with either of two peptides (s or l-protamine-NLS). These peptides bind to and deliver antisense oligonucleotides to the nucleus of T24 bladder and PC3 prostate cancer cells, as demonstrated by confocal microscopy. Furthermore, as shown by Western and Northern blotting, the peptide-oligonucleotide complexes produced excellent downregulation of the expression of the complementary mRNAs, which in turn resulted in downregulation of protein expression. However, under certain circumstances (predominantly in PC3 cells), incubation of the cells with chloroquine was required to produce antisense activity. Using this strategy, PKC alpha protein and mRNA expression in T24 and PC3 cells and bcl-2 expression in PC3 cells was reduced by approximately 75 +/- 10% at a minimum concentration of oligomer of 0.25 microM, in combination with 12-15 microM peptide. On the basis of our results, we conclude that arginine-rich peptides of this class may be potentially useful delivery vehicles for the cellular delivery of antisense oligonucleotides. This new strategy may have several advantages over other methods of oligonucleotide delivery and may complement already existing lipid based technologies. PMID- 11906255 TI - Enzyme-assisted synthesis and structure characterization of glucuronide conjugates of methyltestosterone (17 alpha-methylandrost-4-en-17 beta-ol-3-one) and nandrolone (estr-4-en-17 beta-ol-3-one) metabolites. AB - A new and useful method based on enzyme-assisted synthesis was developed for producing 3 alpha-O-beta-D-glucuronide conjugates from synthetic phase I metabolites of methyltestosterone and nandrolone. The formed glucuronide conjugates of 17 alpha-methyl-5 alpha-androstane-3 alpha,17 beta-diol (I), 17 alpha-methyl-5 beta-androstane-3 alpha,17 beta-diol (II), 5 alpha-estran-3 alpha ol-17-one (III), and 5 beta-estran-3 alpha-ol-17-one (IV) are urinary metabolites, indicating the human misuse of the above-mentioned anabolic androgenic steroids (AAS). The common lack of reference material precludes the use and validation of these biomarkers in human doping control. Liver microsomes from Aroclor 1254-induced rats were used as a highly active source of mammalian UDP-glucuronosyltransferases (UGT, EC 2.4.1.17). After purification by protein precipitation, liquid-liquid extraction (dichloromethane), C-18 solid-phase extraction, and lyophilization, the steroid glucuronide structures were characterized by (1)H and (13)C NMR spectroscopy and tandem mass spectrometry. The enzymatic method was highly stereoselective, producing a single major conjugate from the parent steroids I-IV. The stereochemically pure steroid glucuronide conjugates were recovered in milligram amounts (1.0-2.8 mg, yield 12 29%), which is sufficient for veterinary and human doping control analyses; for pharmaco-, toxico-, and enzyme kinetic studies in the pharmaceutical industry; for clinical laboratories; and for forensic medicine. A new sensitive LC-MS method was developed for controlling the product purity in syntheses, as well as for enzyme kinetic characterization of AAS-metabolizing UGT activities in rat liver toward the aglycones I-IV. In this study, the UGT enzymes responsible for the formation of 3 alpha-O-linked glucuronides from the substrates I, II, III, and IV exhibited the specific enzyme activity values: 25, 124, 48, and 212 nmol/mg microsomal protein in a 2-h incubation, respectively. PMID- 11906256 TI - A homogeneous DNA hybridization system by using a new luminescence terbium chelate. AB - Homogeneous DNA hybridization assay based on the luminescence resonance energy transfer (LRET) from a new luminescence terbium chelate, N,N,N(1),N(1)-[2,6 bis(3'-aminomethyl-1'-pyrazolyl)-4-phenylpyridine]tetrakis(acetic acid) (BPTA) Tb(3+) (lambda(ex) = 325 nm and lambda(em) = 545 nm) to an organic dye, Cy3 (lambda(ex) = 548 nm and lambda(em) = 565 nm), has been developed. In the system, two DNA probes whose sequences are complementary to the two different consecutive sequences of a target DNA are used; one of the probes is labeled with the Tb(3+) chelate at the 3'-end, and the other is with Cy3 at the 5'-end. Labeling of the Tb(3+) chelate is accomplished via the linkage of a biotin-labeled DNA probe with the Tb(3+) chelate-labeled streptavidin. Strong sensitized emission of Cy3 was observed upon excitation of the Tb(3+) chelate at 325 nm, when the two probe DNAs were hybridized with the target DNA. The sensitivity of the assay was very high compared with those of the previous homogeneous-format assays using the conventional organic dyes; the detection limit of the present assay is about 30 pM of the target DNA strand. PMID- 11906257 TI - Star structure of antibody-targeted HPMA copolymer-bound doxorubicin: a novel type of polymeric conjugate for targeted drug delivery with potent antitumor effect. AB - The aim of this study was to compare the properties and antitumor potential of a novel type of antibody-targeted N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-bound doxorubicin conjugates with star structure with those of previously described classic antibody-targeted or lectin-targeted HPMA copolymer bound doxorubicin conjugates. Classic antibody-targeted conjugates were prepared by aminolytic reaction of the multivalent HPMA copolymer containing side-chains ending in 4-nitrophenyl ester (ONp) reactive groups with primary NH(2) groups of the antibodies. The star structure of antibody-targeted conjugates was prepared using semitelechelic HPMA copolymer chains containing only one reactive N hydroxysuccinimide group at the end of the backbone chain. In both types of conjugates, B1 monoclonal antibody (mAb) was used as a targeting moiety. B1 mAb recognizes the idiotype of surface IgM on BCL1 cells. The star structure of the targeted conjugate had a narrower molecular mass distribution than the classic structure. The peak in the star structure was around 300-350 kDa, while the classic structure conjugate had a peak around 1300 kDa. Doxorubicin was bound to the HPMA copolymer via Gly-Phe(D,L)-Leu-Gly spacer to ensure the controlled intracellular delivery. The release of doxorubicin from polymer conjugates incubated in the presence of cathepsin B was almost twice faster from the star structure of targeted conjugate than from the classic one. The star structure of the targeted conjugate showed a lower binding activity to BCL1 cells in vitro, but the cytostatic activity measured by [(3)H]thymidine incorporation was three times higher than that seen with the classic conjugate. Cytostatic activity of nontargeted and anti-Thy 1.2 mAb (irrelevant mAb) modified HPMA copolymer-bound doxorubicin was more than hundred times lower as compared to the star structure of B1 mAb targeted conjugate. In vivo, both types of conjugates targeted with B1 mAb bound to BCL1 cells in the spleen with approximately the same intensity. The classic structure of the targeted conjugate bound to BCL1 cells in the blood with a slightly higher intensity than the star structure. Both types of targeted conjugates had a much stronger antitumor effect than nontargeted HPMA copolymer bound doxorubicin and free doxorubicin. The star structure of targeted conjugate had a remarkably higher antitumor effect than the classic structure: a single intravenous dose of 100 microg of doxorubicin given on day 11 completely cured five out of nine experimental animals whereas the classic structure of targeted conjugate given in the same schedule only prolonged the survival of experimental mice to 138% of control mice. These results show that the star structure of antibody-targeted HPMA copolymer-bound doxorubicin is a suitable conjugate for targeted drug delivery with better characterization, higher cytostatic activity in vitro, and stronger antitumor potential in vivo than classic conjugates. PMID- 11906258 TI - Conjugates bearing multiple formyl-methionyl peptides display enhanced binding to but not activation of phagocytic cells. AB - N-Formyl-methionyl peptides can specifically bind to surface receptors on phagocytic cells. A single copy of N-formyl-methionine-leucine-phenylalanine (fMLF) covalently linked to a poly(ethylene glycol)-based polymer displayed reduced binding avidity (K(d) = 190 nM) for differentiated HL-60 cells relative to free fMLF (K(d) = 28 nM). Increasing the number of fMLF residues (up to eight) attached to a single polymer results in enhanced avidity for these cells (K(d) = 0.18 nM), which appears to be independent of whether the polymer backbone is linear or branched. However, no conjugate showed enhanced ability to activate phagocytic cells, relative to the free peptide (EC(50) = 5 nM), as measured by transient stimulation of release of calcium ions from intracellular stores into the cytoplasm. A polymer bearing four fMLF and four digoxigenin residues showed specific enhancement in binding to differentiated HL-60 cells and mouse peritoneal macrophages in situ relative to a polymer lacking fMLF; no such enhancement was seen in binding to receptor-negative lymphocytic Jurkat cells. These results suggest that multiple fMLF residues linked to a drug-delivery polymer can be used to target appended drugs to phagocytic cells with relatively little toxicity due to cellular activation. PMID- 11906260 TI - Synthesis of sulfhydryl cross-linking poly(ethylene glycol)-peptides and glycopeptides as carriers for gene delivery. AB - Sulfhydryl cross-linking poly(ethylene glycol) (PEG)-peptides and glycopeptides were prepared and tested for spontaneous polymerization by disulfide bond formation when bound to plasmid DNA, resulting in stable PEG-peptide and glycopeptide DNA condensates. A 20 amino acid synthetic peptide possessing a single sulfhydryl group on the N-terminal cysteine, with two or five internal acetamidomethyl (Acm)-protected cysteine residues, was reacted with either PEG vinyl sulfone or iodoacetamide tyrosinamide triantennary N-glycan. Following RP HPLC purification, Acm groups were removed by silver tetrafluoroborate to generate sulfhydryl cross-linking PEG-peptides and glycopeptide that were characterized by either (1)H NMR or LC-MS. Sulfhydryl cross-linking PEG-peptides and glycopeptides were found to bind to plasmid DNA and undergo disulfide cross linking resulting in stable DNA condensates with potential utility for in vivo gene delivery. PMID- 11906259 TI - Chemical synthesis of Escherichia coli ST(h) analogues by regioselective disulfide bond formation: biological evaluation of an (111)In-DOTA-Phe(19)-ST(h) analogue for specific targeting of human colon cancers. AB - New human Escherichia coli heat-stable peptide (ST(h)) analogues containing a DOTA chelating group were synthesized by sequential and selective formation of disulfides bonds in the peptide. This synthetic approach utilizes three orthogonal thiol-protecting groups, Trt, Acm, and t-Bu, to form three disulfide bonds by successive reactions using 2-PDS, iodine, and silyl chloride-sulfoxide systems. The DOTA-ST(h) conjugates exhibiting high guanylin/guanylate cyclase-C (GC-C) receptor binding affinities were obtained with >98% purity. In vitro competitive binding assays, employing T-84 human colon cancer cells, demonstrated the IC(50) values of <2 nM for GC-C receptor binding suggesting that the new synthetic ST(h) analogues are biologically active. In vitro stability studies of the (111)In-DOTA-Phe(19)-ST(h) conjugate incubated in human serum at 37 degrees C under 5% CO(2) atmosphere revealed that this conjugate is extremely stable with no observable decomposition at 24 h postincubation. HPLC analysis of mouse urine at 1 h pi of the (111)In-DOTA-Phe(19)-ST(h) conjugate showed only about 15% decomposition suggesting that the (111)In-DOTA-Phe(19)-ST(h) conjugate is highly stable, even under in vivo conditions. In vivo pharmacokinetic studies of the (111)In-DOTA-Phe(19)-ST(h) conjugate in T-84 human colon cancer derived xenografts in SCID mice conducted at 1 h pi showed an initial tumor uptake of 2.04 +/- 0.30% ID/g at 1 h pi with efficient clearance from the blood pool (0.23 +/- 0.14% ID/g, 1 h pi) by excretion mainly through the renal/urinary pathway (95.8 +/- 0.2% ID, 1 h pi). High tumor/blood, tumor/muscle, and tumor/liver ratios of approximately 9:1, 68:1, and 26:1, respectively, were achieved at 1 h pi The specific in vitro and in vivo uptake of the radioactivity by human colonic cancer cells highlights the potential of radiometalated-DOTA-ST(h) conjugates as diagnostic/therapeutic radiopharmaceuticals. PMID- 11906261 TI - Solid-state NMR investigation of indomethacin-cyclodextrin complexes in PEG 6000 carrier. AB - Various solid dispersions of alpha-, beta- and gamma-cyclodextrin (CD) in PEG 6000 with and without the addition of 5% w/w indomethacin were prepared by the melting method using the original components. The samples were investigated by solid-state (13)C NMR, and the interactions between the drug and the cyclodextrins were evaluated. The indomethacin-gamma-CD phase with tetragonal symmetry found in a previous X-ray study gave chemical shifts which suggested that this phase is a complex between indomethacin and gamma-CD. Evidence of an indomethacin-beta-CD complex were found. A distribution of the chemical shifts for beta-CD was attributed to the possible formation of different types of complexes between indomethacin and beta-CD. No complex formation was found in the alpha-CD system. The degree of relative crystallinity of the samples in the gamma CD system was measured by (1)H NMR, X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), and modulated-temperature DSC (MTDSC). The results obtained by the NMR, XRD, and DSC techniques showed that the dispersions were less crystalline than the pure polymer carrier, and the dispersion containing the indomethacin-gamma-CD complex had the lowest degree of crystallinity. By the MTDSC method a deviation was found for the PEG 6000/indomethacin dispersion. This emphasizes that the different techniques give specific information on the crystallinity. PMID- 11906262 TI - Porphyrin, chlorin, and bacteriochlorin isothiocyanates: useful reagents for the synthesis of photoactive bioconjugates. AB - A novel method for conjugating porphyrins and related molecules to proteins has been developed. The method, which involves synthesizing porphyrins, chlorins, and bacteriochlorins bearing a single amine-reactive isothiocyanate group represents a facile system for protein labeling with these photoactive species. Problems associated with the noncovalent binding of porphyrins to proteins are highlighted, and a method for purifying conjugates to yield exclusively covalently bound porphyrin protein species is demonstrated. Biological activity of porphyrin-bovine serum albumin conjugates formed and purified by these methods is demonstrated using laser scanning confocal microscopy. PMID- 11906263 TI - Tat peptide directs enhanced clearance and hepatic permeability of magnetic nanoparticles. AB - Superparamagnetic nanoparticles have a number of important biomedical applications, serving as MR contrast agents for imaging specific molecular targets, as reagents for cell labeling and cell tracking, and for the isolation of specific classes of cells. We have determined the physical and biological properties of MION-47 and amino-CLIO, nanoparticles which serve as precursors for the synthesis of targeted MR contrast agents, and Tat-CLIO, a nanoparticle used as a cell labeling reagent. Blood half-lives for MION-47 and amino-CLIO were 682 +/- 34 and 655 +/- 37 min, respectively. The attachment of 9.7 tat peptides per crystal to amino-CLIO resulted in a reduction in blood half-life to 47 +/- 6 min. MION-47, amino-CLIO, and Tat-CLIO were present in highest concentrations in liver and spleen and lymph nodes, where concentrations for all three nanoparticles ranged from 8.80 to 6.11% of injected dose per gram. Twenty-four hours after the intravenous injection of amino-CLIO, the nanoparticle was concentrated in cells surrounding hepatic blood vessels (endothelial and Kupffer cells), in a fashion similar to that obtained with other nanoparticle preparations. In contrast, Tat CLIO was present as numerous discrete foci of intense fluorescence throughout the parenchyma. Using the peptide as a component of future nanoparticles, it might be possible to design sensors for the detection of macromolecules present in intracellular compartments. PMID- 11906264 TI - Cysteine-free mutant of aequorin as a photolabel in immunoassay development. AB - The bioluminescent protein aequorin is a sensitive label that has been employed in a number of analytical applications. A mutant of aequorin with enhanced stability produced recombinantly in our laboratory has been employed as a label in the development of an immunoassay for digoxin. Digoxin is a cardiac glycoside used in the treatment of congestive heart failure. This drug has a very narrow therapeutic range of 0.8-2.0 ng/mL (1.0-2.5 nmol/L), thus requiring therapeutic drug monitoring. In this study, a derivative of digoxigenin was chemically conjugated to the mutant aequorin, and the resulting protein-digoxigenin derivative conjugates were characterized in terms of their luminescence properties. A solid-phase immunoassay for digoxin was then developed. The detection limit of the assay for digoxin was 1 x 10(-12) M. To demonstrate the use of this mutant aequorin as a label in biological sample analysis without any need for pretreatment of the samples, the assay was tested in serum spiked with digoxin. Interference from digoxin analogues was also evaluated to determine the specificity of the assay. PMID- 11906265 TI - Noninvasive imaging of peripherally injected Alzheimer's disease type synthetic A beta amyloid in vivo. AB - The pathological hallmark of Alzheimer's disease (AD) is accumulation in the brain of amyloid composed of the 40-mer peptide A beta. Many fundamental questions about the biology of (AD) remain unanswered because there is currently no method of quantifying A beta amyloid in vivo. A noninvasive method of detecting and quantifying A beta amyloid in vivo would have wide application for the premortem diagnosis of AD and the efficient evaluation of candidate therapeutics aimed at inhibiting the formation and growth of A beta amyloid. Taking advantage of the extraordinarily high affinity of A beta for itself, we have synthesized an N'-terminal diethylenetriaminepentaacetic acid (DTPA) derivative of A beta possessing the kinetic activity and specificity for A beta amyloid desired of a probe to be used for noninvasive imaging. DTPA-A beta(3-40) is readily labeled with (111)InOAc(3) to yield a stable probe with exquisite specificity for naturally occurring and synthetic A beta amyloid in vitro. Moreover, (111)In-DTPA-A beta(3-40), administered intravascularly can specifically deposit onto and label previously injected synthetic A beta amyloid and be imaged in vivo with a gamma camera. The present results demonstrate the design, synthesis, and use of an A beta amyloid-specific probe and methods for its use as a noninvasive imaging agent. In vivo imaging of A beta amyloid represents an important step toward the development of biochemically based objective tools for the assessment of progression of AD and efficacy of potential therapeutics. PMID- 11906266 TI - Immobilization of pectin fragments on solid supports: novel coupling by thiazolidine formation. AB - As a prerequisite to solid-phase and sequence analyses and for the study of the fine structure of pectin, we have developed oriented and chemoselective methodologies to couple model pectin fragments onto a solid support. Polyethylene glycol polyacrylamide (PEGA) resins were selected due to their excellent swelling properties in a wide range of solvents, including water, and their easy accessibility to enzymes. Following appropriate derivatization of amino terminated PEGA resins, oligomers of alpha-D-galacturonic acid (GalA), up to the trimer, were anchored to the support through their reducing end. In addition to reductive amination, the strategies included the formation of an oxime bond, a glycosyl hydrazide, and a pyroglutamyl ring. Further, we developed a new immobilization approach based on the formation of a thiazolidine ring. All methods proved efficient and did not require modification of the GalA oligomers prior to coupling. In addition, very mild conditions and few steps for derivatization of the support were required. Immobilization by thiazolidine ring and oxime bond formation were the preferred methods, given the stability of the linkages formed, their compatibility with aqueous solvents, the few number of steps required, and their potential for application to larger pectin fragments. Thiazolidine and pyroglutamyl anchoring were developed further by the insertion of a disulfide bond which allowed release of the saccharides under mild, selective conditions. PMID- 11906267 TI - Design of a targeted peptide nucleic acid prodrug to inhibit hepatic human microsomal triglyceride transfer protein expression in hepatocytes. AB - In this study, we present the design and synthesis of an antisense peptide nucleic acid (asPNA) prodrug, which displays an improved biodistribution profile and an equally improved capacity to reduce the levels of target mRNA. The prodrug, K(GalNAc)(2)-asPNA, comprised of a 14-mer sequence complementary to the human microsomal triglyceride transfer protein (huMTP) gene, conjugated to a high affinity tag for the hepatic asialoglycoprotein receptor (K(GalNAc)(2)). The prodrug was avidly bound and rapidly internalized by HepG2s. After iv injection into mice, K(GalNAc)(2)-asPNA accumulated in the parenchymal liver cells to a much greater extent than nonconjugated PNA (46% +/- 1% vs 3.1% +/- 0.5% of the injected dose, respectively). The prodrug was able to reduce MTP mRNA levels in HepG2 cells by 35-40% (P < 0.02) at 100 nM in an asialoglycoprotein receptor- and sequence-dependent fashion. In conclusion, hepatocyte-targeted PNA prodrugs combine a greatly improved tropism with an enhanced local intracellular availability and activity, making them attractive therapeutics to lower the expression level of hepatic target genes such as MTP. PMID- 11906268 TI - Preparation of a conjugate of 2',5'-triadenylate 5'-triphosphate and biotinylated firefly luciferase and its use for sensitive bioluminescent enzyme immunoassay. AB - A bioluminescent enzyme immunoassay (BLEIA) for 2',5'-oligoadenylate 5' triphosphate (pppA2'p5'A2'Ap5'A, 2-5A) was developed using a conjugate of 2-5A and firefly luciferase as a probe. The conjugate was prepared from a 2-5A derivative bearing a thiol group at the 2'(or 3') end, and streptavidin (SA) bearing maleimide via a linker-arm and biotinylated luciferase. The thiol group of the 2-5A derivative was relatively stable under aerobic conditions and remained 100% and 56% intact at 25 degrees C after 1 and 96 h, respectively, without dimerization by aerobic oxidation. The thiol-modified 2-5A was coupled with the SA-maleimide conjugate, followed by complex formation with biotinylated firefly luciferase. BLEIA using this conjugate allowed for the direct analysis of 2-5A in a range of 5-500 fmol (0.1-10 nM in a 50 microL sample) and in a reaction time of 15 min. The measurement of microquantities of 2-5 A from various sources is easy to perform by this BLEIA, because no radioisotope labeled 2-5A was required as a probe. PMID- 11906269 TI - Synthesis of new modified DNAs by hyperthermophilic DNA polymerase: substrate and template specificity of functionalized thymidine analogues bearing an sp3 hybridized carbon at the C5 alpha-position for several DNA polymerases. AB - Novel thymidine analogue triphosphates, which have an sp3-hybridized carbon at the C5 alpha-position with amino-linker arms, a methyl ester, or a carboxyl group at the C5 sidearm, were good substrates for primer-extension reactions by DNA polymerase from Pyrococcus kodakaraensis (KOD Dash DNA polymerase), yielding exclusively full-length products. The resulting modified DNA was further allowed to react with a functional molecule such as fluorescein isothiocyanate. By contrast, only truncated products were formed from the thymidine analogue substrate bearing the amino-linker arm or the negatively charged carboxyl group using Taq, Tth DNA polymerase, or DNA polymerase I from E. coli (Klenow fragment). The results indicate either that the thymidine analogue was not accepted by the enzymes, or that the polymerases could not extend the products, once the analogue had been incorporated, depending on the type of the analogue. A conventional thymidine analogue bearing an aminopropenyl group at the C5-position was accepted by all enzymes, among which KOD Dash DNA polymerase showed the highest activity for the polymerization with this analogue. Templates bearing the thymidine analogues in place of one thymidine residue were read by KOD Dash, Taq, Tth DNA polymerases, and the Klenow fragment giving the full-length product. KOD Dash DNA polymerase could expand structural diversities of substrates that can be used to prepare modified DNAs. PMID- 11906271 TI - Synthesis of the first diethylenetriaminepentahydroxamic acid (DTPH) bifunctional chelating agent. AB - Synthesis of a new pentahydroxamic acid bifunctional chelating agent (BCA), constructed on the aminoazaalkyl core of diethylenetriaminepentaacetic acid (DTPA), is reported. Rational modifications in the structure of DTPA, which could result in an enhancement of its chelation properties, add to the collection of diagnostic and therapeutic metals bound by this chelator, and might implement significant improvements in the in vivo behavior of this compound, are described. Further improvements in the stability of the ligand-metal complexes of DTPA may improve both diagnostic and therapeutic outcomes such as tumor-to-normal tissue ratios and target-delivered radioactivity. A combination of hydroxamate functions with the azaalkyl backbone of DTPA might be a suitable approach to generate such higher stabilities. This rationale may be justified by the well-known affinity of hydroxamates against different transition metals and favorable properties of DTPA as a versatile chelator. Thus, the N(4),N(alpha),N(alpha),N(epsilon),N(epsilon) pentakis[[((N-hydroxy-N-methyl]carbonyl)methyl]-2, 6-diamino-4-azahexanoic hydrazide (5, DTPH) was designed and synthesized through a convergent synthesis and in 40.7% overall yield. Conjugation of this compound to the monoclonal antibody (MAb) Delta Ch2HuCC49, used as a model protein, was carried out to evaluate the efficiency of this molecule as a BCA. Radiolabeling of the DTPH Delta CH2HuCC49 conjugate with lutetium-177 ((177)Lu) and biodistribution of the labeled conjugate in athymic nude mice, bearing LS174T human colon carcinoma xenografts, are reported. PMID- 11906270 TI - De novo synthesis of a new diethylenetriaminepentaacetic acid (DTPA) bifunctional chelating agent. AB - Diethylene triamine pentaacetic acid (DTPA) has been in extensive use as a metal chelator in the development of radiopharmaceuticals and contrast agents. The former application uses DTPA mostly as a bifunctional chelating agent (BCA) conjugated to tumor-targeting vehicles (TTVs) such as monoclonal antibodies (MAbs) and receptor-directed peptides. A new bifunctional DTPA derivative was synthesized by a fully organic scheme. This compound, N(4),N(alpha),N(alpha),N(epsilon),N(epsilon)-[pentakis(carboxymethyl)]-N(4) (carboxymethyl)-2,6-diamino-4-azahexanoic hydrazide (20) was prepared by a convergent synthesis strategy using N(alpha)-benzyloxycarbonyl-2,3 diaminopropionic acid as the starting compound. This commercially available material was used to build a functionalized triamine which served as the molecular core template for assembling the target molecule. To evaluate the conjugation and radiolabeling capabilities of this new molecule, it was covalently attached to the anti-TAG-72 MAb, Delta CH2HuCC49, and the conjugate was radiolabeled in near-quantitative yields with yttrium-90 ((90)Y) and lutetium 177 ((177)Lu). Biodistribution of the (177)Lu-labeled DTPA-Delta CH2HuCC49 in tumor-bearing nude mice demonstrated preservation of the immunoreactivity of the MAb as indicated by high tumor uptake. In addition to the introduction of a new bifunctional DTPA, this work reports on a novel synthetic approach for preparation of this useful metal chelator and introduces a new conjugation protocol. PMID- 11906272 TI - Synthesis and cleavage experiments of oligonucleotide conjugates with a diimidazole-derived catalytic center. AB - Ribonuclease mimics based on diimidazole derived constructs in combination with or without additional amino groups have been synthesized and conjugated to oligonucleotides. The imidazole moiety was used either unprotected, protected with a monomethoxytrityl group or a tert-butyloxy carbonyl group. Acylation reactions were carried out using the 3-acyl-1,3-thiazolidine-2-thione activation strategy. The peptides were coupled to the oligonucleotides with a mixture of PyBOP, DIEA an HOBt in DMF on solid support. The conjugates were purified by RP HPLC and identified using negative ion mode mass spectrometry. Unfortunately, no cleavage of a linear RNA target under physiological conditions could be observed. PMID- 11906273 TI - Molecular umbrella-assisted transport of thiolated AMP and ATP across phospholipid bilayers. AB - Two molecular umbrella-nucleoside conjugates (1a and 1b) have been synthesized via thiolate-disulfide displacement by adenosine 5'-O-(3-thiomonophosphate) and adenosine 5'-O-(3-thiotriphosphate) on an activated dimer derived from cholic acid, spermidine, and 5,5'-dithiobis-(2-nitrobenzoic acid). Both conjugates readily enter the aqueous compartment of liposomes made from 1-palmitoyl-2-oleyol sn-glycero-3-phosphocholine (POPC) and release the free nucleoside upon reaction with entrapped glutathione. Approximately 50% of the thiolated form of AMP is released within 20 min at 23 degrees C; 120 min is required for a similar release of the thiolated form of ATP. The facile cleavage of these conjugates by glutathione, together with the fact that mammalian cells contain millimolar concentrations of this tripeptide in their cytoplasm, suggest that such chemistry may be extended to the practical development of prodrugs, e.g., antisense oligonucleotides that can be delivered into cells. PMID- 11906274 TI - Synthesis and biologic evaluation of a radioiodinated quinazolinone derivative for enzyme-mediated insolubilization therapy. AB - We have developed a new strategy that aims to concentrate therapeutic radionuclides within solid tumors. This approach, which we have named EMIT (enzyme-mediated insolubilization therapy), is a method for enzyme-dependent, site-specific, in vivo precipitation of a radioactive molecule (from a water soluble precursor) within the extracellular space of solid tumors. The prodrug, ammonium 2-(2'-phosphoryloxyphenyl)-6-iodo-4-(3H)-quinazolinone, labeled with iodine-125 ((125)IPD) and its authentic compound labeled with iodine-127 (IPD) have been synthesized, purified, and characterized. The alkaline phosphatase (ALP)-mediated conversion of these water-soluble nonfluorescent prodrugs to the water-insoluble fluorescent species, iodine-125-labeled 2-(2'-hydroxyphenyl)-6 iodo-4-(3H)-quinazolinone ((125)ID) and its iodine-127-labeled derivative (ID), has been demonstrated in vitro. Biodistribution studies in mice indicate that both (125)IPD and (125)ID are minimally retained by most tissues and organs. In addition, following its intravenous injection in mice, (125)IPD is localized in ALP-rich regions and converted to (125)ID, which remains indefinitely within the tissues where it is produced. We believe that EMIT is a strategy that will lead to the active and specific concentration and entrapment of therapeutic radionuclides within solid tumors, the consequent protracted irradiation of tumor cells within the range of the emitted particles, and the effective therapy of solid tumors. PMID- 11906275 TI - Conjugation of various acridines to DNA for site-selective RNA scission by lanthanide ion. AB - Three types of DNA conjugates having 9-acridinecarboxamide, 9-aminoacridine, and 9-amino-6-chloro-2-methoxyacridine at the 5'-ends were synthesized and used for site-selective RNA scission together with another unmodified DNA and Lu(III) ion. The target phosphodiester linkages in the substrate RNA were selectively and efficiently activated and were hydrolyzed by free Lu(III) ion. The conjugate bearing 9-amino-6-chloro-2-methoxyacridine was the most active. However, its duplex with the substrate RNA was almost as stable as that of the 9-aminoacridine bearing conjugate, which was much less active for the RNA activation. The 9 acridinecarboxamide-bearing conjugate was only marginally active. The substituents on the acridine groups in these conjugates positively participate in the present RNA activation, probably by fixing the orientation of the acridine rings. PMID- 11906276 TI - Effects of coligand variation on the in vivo characteristics of Tc-99m-labeled interleukin-8 in detection of infection. AB - In our previous studies, interleukin-8 (IL-8) was labeled with (99m)Tc using hydrazinonicotinamide (HYNIC) as bifunctional coupling agent and tricine as coligand. This preparation showed excellent characteristics for imaging of infection in a rabbit model of soft-tissue infection. In the present study, the propylaldehyde hydrazone formulation of HYNIC was introduced to stabilize HYNIC IL-8. (99m)Tc-HYNIC-IL-8 was prepared using 5 different coligand formulations. The effect of these coligand formulations on the in vitro characteristics and in vivo behavior of (99m)Tc-HYNIC-IL-8 was investigated. HYNIC-conjugated IL-8 was labeled with (99m)Tc in the presence of either (A) tricine, (B) ethylenediaminediacetic acid (EDDA), (C) tricine and trisodium triphenylphosphinetrisulfonate (TPPTS), (D) tricine and nicotinic acid (NIC), or (E) tricine and isonicotinic acid (ISONIC). These preparations were characterized in vitro by RP-HPLC, determination of the octanol/water partition coefficient, stability studies, and receptor binding assays. The in vivo biodistribution of the radiolabel in rabbits with E. coli-induced soft-tissue infection was determined both by gamma-camera imaging as well as by tissue counting at 6 h pi. Specific activity (MBq/microg) was highest for (ISO)NIC (up to 80) > TPPTS (40) > tricine (15) > EDDA (7). RP-HPLC and octanol/water partition coefficients showed a shift toward higher lipophilicity for the TPPTS preparation. The leukocyte receptor binding fractions were around 40-55% for all preparations except for TPPTS, which showed predominantly nonspecific binding. All preparations were stabilized in serum, but the stability in PBS was highest for NIC and TPPTS > EDDA > ISONIC > tricine. The in vivo biodistribution showed highest abscess/muscle for NIC and ISONIC (>200) > EDDA and tricine (approximately 100) > TPPTS (<40). Gamma camera imaging rapidly visualized the abscess from 2 h pi onward for all formulations. The abscess/background (A/B) at 6 h pi for ISONIC was significantly higher (P < 0.05) than that of tricine and the A/B of TPPTS was significantly lower (P < 0.05). IL-8 can be rapidly and easily labeled with (99m)Tc using HYNIC as a chelator in combination with various coligands. The most optimal infection imaging characteristics were found for formulations using nicotinic acid/tricine as coligand system combining a high specific activity and high in vitro stability with high abscess/muscle ratios (>200) and high abscess/background ratios (>20). Protein doses to be administered were as low as 70 ng/kg bodyweight. At these low protein doses, side effects are not to be expected in the human system. This paves the way for infection imaging studies in patients. PMID- 11906277 TI - Advantage of the ether linkage between the positive charge and the cholesteryl skeleton in cholesterol-based amphiphiles as vectors for gene delivery. AB - Twelve novel cationic cholesterol derivatives with different linkage types between the cationic headgroup and the cholesteryl backbone have been developed. These have been tested for their efficacies as gene transfer agents as mixtures with dioleoyl phosphatidylethanolamine (DOPE). A pronounced improvement in transfection efficiency was observed when the cationic center was linked to the steroid backbone using an ether type bond. Among these, cholest-5-en-3b-oxyethane N,N,N-trimethylammonium bromide (2a) and cholest-5-en-3b-oxyethane-N,N-dimethyl-N 2-hydroxyethylammonium bromide (3d) showed transfection efficiencies considerably greater than commercially available reagents such as Lipofectin or Lipofectamine. To achieve transfection, 3d did not require DOPE. Increasing hydration at the headgroup level for both ester- and ether-linked amphiphiles resulted in progressive loss of transfection efficiency. Transfection efficiency was also greatly reduced when a 'disorder'-inducing chain like an oleyl (cis-9 octadecenyl) segment was added to these cholesteryl amphiphiles. Importantly, the transfection ability of 2a with DOPE in the presence of serum was significantly greater than for a commercially available reagent, Lipofectamine. This suggests that these novel cholesterol-based amphiphiles might prove promising in applications involving liposome-mediated gene transfection. This investigation demonstrates the importance of structural features at the molecular level for the design of cholesterol-based gene delivery reagents that would aid the development of newer, more efficient formulations based on this class of molecules. PMID- 11906279 TI - Design, synthesis, and evaluation of opioid analogues with non-peptidic beta-turn scaffold: enkephalin and endomorphin mimetics. AB - We have identified a mu-selective opioid receptor agonist without a cationic amino group in the molecule from libraries of bicyclic beta-turn peptidomimetics. The biologically active conformation of the lead is proposed to mimic an endomorphin type III 4 --> 1 beta-turn conformation. PMID- 11906280 TI - Toward selective ERbeta agonists for central nervous system disorders: synthesis and characterization of aryl benzthiophenes. AB - In an effort to identify selective ligands for the estrogen receptor subtype ERbeta, a series of aryl benzthiophenes was synthesized. In a radioligand binding assay and reporter gene assays in HeLa and SH-SY5Y cells, compounds were characterized as ERbeta-selective agonists. By targeting ERbeta in the brain, these compounds could lead to drugs able to separate the beneficial effects of estrogens on mood, learning, and memory from side effects such as the stimulation of endometrial and breast cancer. PMID- 11906281 TI - Identification of novel cyclooxygenase-2 selective inhibitors using pharmacophore models. AB - In the present study we have investigated whether pharmacophore models may account for the activity and selectivity of the known cyclooxygenase-2 (COX-2) selective inhibitors of the phenylsulfonyl tricyclic series, i.e., Celecoxib (1) and Rofecoxib (3), and whether transferring this structural information onto the frame of a nonsteroidal antiinflammatory drug (NSAID), known to tightly bind the enzyme active site, may be useful for designing novel COX-2 selective inhibitors. With this aim we have developed a pharmacophore based on the geometric disposition of chemical features in the most favorable conformation of the COX-2 selective inhibitors SC-558 (2; analogue of Celecoxib (1)) and Rofecoxib (3) and the more restrained compounds 4 (DFU) and 5. The pharmacophore model contains a sulfonyl S atom, an aromatic ring (ring plane A) with a fixed position of the normal to the plane, and an additional aromatic ring (ring plane B), both rings forming a dihedral angle of 290 degrees +/- 10 degrees. The final disposition of the pharmacophoric groups parallels the geometry of the ligand SC-558 (2) in the known crystal structure of the COX-2 complex. Moreover, the nonconserved residue 523 is known to be important for COX-2 selective inhibition; thus, the crystallographic information was used to position an excluded volume in the pharmacophore, accounting for the space limits imposed by this nonconserved residue. The geometry of the final five-feature pharmacophore was found to be consistent with the crystal structure of the nonselective NSAID indomethacin (6) in the COX-2 complex. This result was used to design indomethacin analogues 8 and 9 that exhibited consistent structure-activity relationships leading to the potent and selective COX-2 inhibitor 8a. Compound 8a (LM-1685) was selected as a promising candidate for further pharmacological evaluation. PMID- 11906282 TI - Molecular dynamics and free energy analyses of cathepsin D-inhibitor interactions: insight into structure-based ligand design. AB - In this study, we compare the calculated and experimental binding free energies for a combinatorial library of inhibitors of cathepsin D (CatD), an aspartyl protease. Using a molecular dynamics (MD)-based, continuum solvent method (MM PBSA), we are able to reproduce the experimental binding affinity for a set of seven inhibitors with an average error of ca. 1 kcal/mol and a correlation coefficient of 0.98. By comparing the dynamical conformations of the inhibitors complexed with CatD with the initial conformations generated by CombiBuild (University of California, San Francisco, CA, 1995), we have found that the docking conformation observed in an X-ray structure of one peptide inhibitor bound to CatD (Proc. Natl. Acad. Sci. U.S.A. 1993, 90, 6796-6800) is in good agreement with our MD simulation. However, the DOCK scoring function, based on intermolecular van der Waals and electrostatics, using a distance-dependent dielectric constant (J. Comput. Chem. 1992, 13, 505-524), poorly reproduces the trend of experimental binding affinity for these inhibitors. Finally, the use of PROFEC (J. Comput.-Aided Mol. Des. 1998, 12, 215-227) analysis allowed us to identify two possible substitutions to improve the binding of one of the better inhibitors to CatD. This study offers hope that current methods of estimating the free energy of binding are accurate enough to be used in a multistep virtual screening protocol. PMID- 11906283 TI - Studies on the mechanism of hypoxic selectivity in copper bis(thiosemicarbazone) radiopharmaceuticals. AB - Copper diacetyl-bis(N4-methylthiosemicarbazone), Cu(II)ATSM, is a promising agent for imaging hypoxic tissue. Here we present results that provide insight into the chemical and electronic properties underlying previously observed structure activity relationships. Density functional theory (DFT) calculations on the electronic structures and molecular orbitals of a series of 13 Cu(II)bis(thiosemicarbazone) analogues with different alkylation patterns and with fixed geometries based on the known structure of Cu(II)PTSM showed that the LUMO and the next lowest orbital were very close in energy, and their energy order was strikingly dependent on the ligand alkylation pattern in a way that correlated with hypoxia-selectivity and redox potentials. The LUMOs of Cu(II)ATSM and other hypoxia-selective analogues were predominantly metal-based (leading to a singlet reduced species) while the LUMOs of Cu(II)PTSM and other nonselective analogues were predominantly ligand-based (leading to a triplet reduced species). Upon relaxation of the geometric constraint and full optimization in both Cu(II)ATSM and Cu(II)GTS, the metal-based orbital became the LUMO, and the singlet was the thermodynamically preferred form of the reduced species. Chemical and electrochemical investigation showed that all Cu(II) complexes were reducible, but Cu(I)PTSM and other nonselective analogues dissociated immediately upon reduction with release of ligand (detected by UV-vis) while Cu(I)ATSM and other hypoxia-selective analogues did not. Instead they were rapidly re-oxidized to the Cu(II) complex by molecular oxygen. The reversible electrochemical reduction of nonselective complexes Cu(II)PTSM and Cu(II)GTS became irreversible in the presence of weak acid, whereas that of Cu(II)ATSM was unaffected. In light of these results we present a model to explain the structure-activity relationships on the basis of electronic structure and molecular vibrations. PMID- 11906284 TI - Hydroxyethylamine isostere of an HIV-1 protease inhibitor prefers its amine to the hydroxy group in binding to catalytic aspartates. A synchrotron study of HIV 1 protease in complex with a peptidomimetic inhibitor. AB - A complex structure of HIV-1 protease with a hydroxyethylamine-containing inhibitor Boc-Phe-Psi[(S)-CH(OH)CH2NH]-Phe-Gln-Phe-NH2 has been determined by X ray diffraction to 1.8 A resolution. The inhibitor is bound in the active site of the protease dimer with its hydroxyethylamine isostere participating in hydrogen bonds to the catalytic aspartates 25 and 25' and glycine 27' of the active site triads via five hydrogen bonds. The isostere amine interactions with the catalytic aspartates result in a displacement of the isostere hydroxy group in comparison with the common position known for analogous hydroxyethylamine containing inhibitors. A comparison with another inhibitor of this series shows that the change of one atom of the P2' side chain (Glu/Gln) leads to an altered ability of creating hydrogen bonds to the active site and within the inhibitor molecule. The diffraction data collected at a synchrotron radiation source enabled a detailed analysis of the complex solvation and of alternative conformations of protein side chains. PMID- 11906285 TI - Structural basis for the glucocorticoid response in a mutant human androgen receptor (AR(ccr)) derived from an androgen-independent prostate cancer. AB - The crystal structure of a mutant androgen receptor (AR) ligand-binding domain (LBD) in complex with the agonist 9alpha-fluorocortisol has been determined at 1.95 A resolution. This mutant AR contains two mutations (L701H and T877A) and was previously reported as a high-affinity cortisol/cortisone responsive AR (AR(ccr)) isolated from the androgen-independent human prostate cancer cell lines MDA PCa 2a and 2b (Zhao et al. Nature Med. 2000, 6, 703-6). The three-dimensional structure of the AR(ccr) LBD complexed with 9alpha-fluorocortisol shows the typical conformation of an agonist-bound nuclear receptor in which helix 12 is precisely positioned as a "lid" for the ligand-binding pocket. Binding of 9alpha fluorocortisol to the AR(ccr) involves favorable hydrogen bond patterns on the C17 and C21 substituents of the ligand due to the mutations at 701 and 877 in the AR(ccr). Our studies provide the first structural explanation for the glucocorticoid activation of AR(ccr), which is important for the development of new therapeutic treatments for androgen-independent prostate cancer. PMID- 11906286 TI - Molecular interaction of the antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1- (2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide with the CB1 cannabinoid receptor. AB - N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole 3-carboxamide (SR141716; 1) is a potent and selective antagonist for the CB1 cannabinoid receptor. Using the AM1 molecular orbital method, conformational analysis of 1 around the pyrazole C3 substituent identified four distinct conformations designated Tg, Ts, Cg, and Cs. The energetic stability of these conformers followed the order Tg > Cg > Ts > Cs for the neutral (unprotonated) form of 1 and Ts > Tg > Cs > Cg for its piperidine N-protonated form. Unified pharmacophore models for the CB1 receptor ligands were developed by incorporating the protonated form of 1 into the superimposition model for the cannabinoid agonists 4-[4-(1,1-dimethylheptyl)-2-hydroxyphenyl]perhydro-2alpha,6beta dihydroxynaphthalene (CP55244; 2) and the protonated form of (R)-[2,3-dihydro-5 methyl-3-[(4-morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl](1 naphthalenyl)methanone (WIN55212-2; 3) reported previously (Shim et al. In Rational Drug Design Symposium Series; Parrill, A. L., Reddy, M. R., Eds.; American Chemical Society: Washington, DC, 1999; pp 165-184). Values of K(i) for 1 and a series of 31 structural analogues were determined from radioligand binding analyses by competitive displacement of [3H]CP55940 from cannabinoid receptors in a rat brain membrane preparation. Comparative molecular field analysis (CoMFA) was employed to construct three-dimensional (3D)-quantitative structure-activity relationship (QSAR) models for this data set as unprotonated species assuming the Tg, Cg, and Ts conformers and for the protonated species assuming the Ts, Tg, and Cs conformers. Values of the conventional r2 and cross validated r2 (r(cv)2) associated with these CoMFA models exceeded the threshold for statistical robustness (r2 > or = 0.90) and internal predictive ability (r(cv)2 > or = 0.50) in each of these six cases except for the protonated species assuming the Tg conformer (i.e., r2 = 0.97; r(cv)2 = 0.36). Results from conformational analyses, superimposition models, and 3D-QSAR models suggest that the N1 aromatic ring moiety of 1 dominates the steric binding interaction with the receptor in much the same way as does the C3 alkyl side chain of cannabinoid agonists and the C3 aroyl ring of the aminoalkylindole agonists. We also determined that several of the conformers considered in this study possess the proper spatial orientation and distinct electrostatic character to bind to the CB1 receptor. We propose that the unique region in space occupied by the C5 aromatic ring of 1 might contribute to conferring antagonist activity. We further propose that the pyrazole C3 substituent of 1 might contribute to conferring either neutral antagonist or inverse agonist activity, depending upon the interaction with the receptor. PMID- 11906287 TI - Modeling of binding modes and inhibition mechanism of some natural ligands of farnesyl transferase using molecular docking. AB - Several natural inhibitors of farnesyl transferase have been reported in the literature: some compounds are competitive with farnesyl pyrophosphate (FPP), whereas other ones are competitive with Ras proteins, even though it is usually hard to highlight their inhibition mechanism, which is still unknown for several natural compounds. The aim of this work is to show that the molecular docking analysis can be successfully used to underline the inhibition mechanism of these natural compounds. First, the selected compounds were subjected to a detailed docking analysis, by means of BioDock, a program able to reveal the most likely binding mode for each ligand. By comparing these results with the binding sites for the natural substrates, earlier determined, it was possible to highlight the site specificity and the inhibition mechanism of the selected compounds. In addition, it is possible to relate the binding mode of these molecules with their lipole values, which is appreciably less for peptidomimetics than for FPP mimetic and reveals a straightforward method to predict and to understand the inhibition mechanism of these natural derivatives. PMID- 11906289 TI - Toward an optimal joint recognition of the S1' subsites of endothelin converting enzyme-1 (ECE-1), angiotensin converting enzyme (ACE), and neutral endopeptidase (NEP). AB - The formation of vasoconstrictors (e.g., angiotensin II and endothelin) and the inactivation of vasodilators (e.g., bradykinin and atrial natriuretic) by membrane-bound zinc metallopeptidases are key mechanisms in the control of blood pressure and fluid homeostasis. The way in which these peptides modulate physiological functions has been intensively studied. With the aim to develop compounds that can jointly block the three metallopeptidases-neutral endopeptidase (NEP, neprilysin), angiotensin-converting enzyme (ACE), and endothelin-converting enzyme (ECE-1)-we studied the common structural specificity of the S1' subsites of these peptidases. Various mercaptoacyl amino acids of the general formula HS-CH2-CH(R1')CO-Trp-OH, possessing more or less constrained R1' side chains, were designed. The mercapto-acyl synthons contain one or two asymmetrical centers. The K(i) values of the separated stereoisomers of the most efficient inhibitors were used to determine the stereochemical preference of each enzyme. A guideline for the joint inhibition of the three peptidases was obtained with the (2R,3R) isomer of compound 13b. Its K(i) values on NEP, ACE, and ECE were 0.7, 43, and 26 nM, respectively. PMID- 11906288 TI - Carbonic anhydrase inhibitors. A general approach for the preparation of water soluble sulfonamides incorporating polyamino-polycarboxylate tails and of their metal complexes possessing long-lasting, topical intraocular pressure-lowering properties. AB - Reaction of polyamino-polycarboxylic acids or their dianhydrides with aromatic/heterocyclic sulfonamides possessing a free amino/imino/hydrazino/hydroxy group afforded mono- and bis-sulfonamides containing polyamino-polycarboxylic acid moieties in their molecule. The acids/anhydrides used in synthesis included IDA, NTA, EDDA, EDTA and EDTA dianhydride, DTPA and DTPA dianhydride, EGTA and EGTA dianhydride, and EDDHA, among others. All the newly prepared derivatives showed strong affinity toward isozymes I, II, and IV of carbonic anhydrase (CA). Metal complexes of the new compounds have also been prepared. Metal ions used in such preparations included di- and trivalent main-group and transition cations, such as Zn(II), Cu(II), Al(III), etc. Some of the new sulfonamides/disulfonamides obtained in this way, as well as their metal complexes, behaved as nanomolar CA inhibitors against isozymes II and IV, being slightly less effective in inhibiting isozyme I. Some of these sulfonamides as well as their metal complexes strongly lowered intraocular pressure (IOP) when applied topically, directly into the normotensive/glaucomatous rabbit eye, as 1-2% water solutions/suspensions. The good water solubility of these sulfonamide CA inhibitors, correlated with the neutral pH of their water solutions used in the ophthalmologic applications and the long duration of action of the IOP-lowering effect, makes them interesting candidates for developing novel types of antiglaucoma drugs devoid of serious topical side effects. PMID- 11906290 TI - Synthesis and structure-activity relationships of 3-aryloxindoles: a new class of calcium-dependent, large conductance potassium (maxi-K) channel openers with neuroprotective properties. AB - A series of 3-aryloxindole derivatives were synthesized and evaluated as activators of the cloned maxi-K channel mSlo expressed in Xenopus laevis oocytes using electrophysiological methods. The most promising maxi-K openers to emerge from this study were (+/-)-3-(5-chloro-2-hydroxyphenyl)-1,3-dihydro-3-hydroxy-6 (trifluoromethyl)-2H-indol-2-one ((+/-)-8c) and its 3-des-hydroxy analogue (+/-) 11b. The individual enantiomers of (+/-)-8c were synthesized, and the maxi-K channel-opening properties were shown to depend on the absolute configuration of the single stereogenic center with the efficacy of (-)-8c superior to that of both (+)-8c and the racemic mixture when evaluated at a concentration of 20 microM. Racemic 11b exhibited greater efficacy than either the racemic 8c or the more active enantiomer in the electrophysiological evaluation. In vitro metabolic stability studies conducted with (+/-)-8c and (+/-)-11b in rat liver S9 microsomal fractions revealed significant oxidative degradation with two hydroxylated metabolites observed by liquid chromatography/mass spectrometry for each compound in addition to the production of 8c from 11b. The pharmacokinetic properties of (+/-)-8c and (+/-)-11b were determined in rats as a prelude to evaluation in a rat model of stroke that involved permanent occlusion of the middle cerebral artery (MCAO model). In the MCAO model, conducted in the spontaneously hypertensive rat, the more polar 3-hydroxy derivative (+/-)-8c did not demonstrate a significant reduction in cortical infarct volume when administered intravenously at doses ranging from 0.1 to 10 mg/kg as a single bolus 2 h after middle cerebral artery occlusion when compared to vehicle-treated controls. In contrast, intravenous administration of (+/-)-11b at a dose of 0.03 mg/kg was found to reduce the measured cortical infarct volume by approximately 18% when compared to vehicle-treated control animals. Intraperitoneal administration of (+/-)-11b at a dose of 10 mg/kg 2 h following artery occlusion was shown to reduce infarct volume by 26% when compared to vehicle-treated controls. To further probe the effects of compounds (+/-)-8c and (+/-)-11b on neurotransmitter release in vitro, both compounds were examined for their ability to reduce electrically stimulated [3H]-glutamate release from rat hippocampal slices that had been preloaded with [3H]-glutamate. Only (+/-)-11b was able to demonstrate a significant inhibition [3H]-glutamate release in this assay at a concentration of 20 microM, providing concordance with the profile of these compounds in the MCAO model. Although (+/-)-11b showed some promise as a potential developmental candidate for the treatment of the sequelae of stroke based on its efficacy in the rat MCAO model, the pharmacokinetic profile of this compound was considered to be less than optimal and was not pursued in favor of derivatives with enhanced metabolic stability. PMID- 11906292 TI - 4-(4-cycloalkyl/aryl-oxazol-5-yl)benzenesulfonamides as selective cyclooxygenase 2 inhibitors: enhancement of the selectivity by introduction of a fluorine atom and identification of a potent, highly selective, and orally active COX-2 inhibitor JTE-522(1). AB - A series of 4-(4-cycloalkyl/aryl-oxazol-5-yl)benzenesulfonamide derivatives were synthesized and evaluated for their abilities to inhibit cyclooxygenase-2 (COX-2) and cyclooxygenase-1 (COX-1) enzymes. In this series, substituent effects at the ortho position to the sulfonamide group on the phenyl ring were examined. Most substituents reduced or lost both COX-2 and COX-1 activities. In contrast, introduction of a fluorine atom preserved COX-2 potency and notably increased COX1/COX-2 selectivity. This work led to the identification of a potent, highly selective, and orally active COX-2 inhibitor JTE-522 [9d, 4-(4-cyclohexyl-2 methyloxazol-5-yl)-2-fluorobenzenesulfonamide], which is currently in phase II clinical trials for the treatment of rheumatoid arthritis, osteoarthritis, and acute pain. PMID- 11906291 TI - 1,8-disubstituted xanthine derivatives: synthesis of potent A2B-selective adenosine receptor antagonists. AB - 3-Unsubstituted xanthine derivatives bearing a cyclopentyl or a phenyl residue in the 8-position were synthesized and developed as A2B adenosine receptor antagonists. Compounds bearing polar substituents were prepared to obtain water soluble derivatives. 1-Alkyl-8-phenylxanthine derivatives were found to exhibit high affinity for A2B adenosine receptors (ARs). 1,8-disubstituted xanthine derivatives were equipotent to or more potent than 1,3,8-trisubstituted xanthines at A2B ARs, but generally less potent at A1 and A2A, and much less potent at A3 ARs. Thus, the new compounds exhibited increased A2B selectivity versus all other AR subtypes. 9-Deazaxanthines (pyrrolo[2,3-d]pyrimidindiones) appeared to be less potent at A2B ARs than the corresponding xanthine derivatives. 1-Propyl-8-p sulfophenylxanthine (17) was the most selective compound of the present series, exhibiting a K(i) value of 53 nM at human A2B ARs and showing greater than 180 fold selectivity versus human A1 ARs. Compound 17 was also highly selective versus rat A1 ARs (41-fold) and versus the other human AR subtypes (A2A > 400 fold and A3 > 180-fold). The compound is highly water-soluble due to its sulfonate function. 1-Butyl-8-p-carboxyphenylxanthine (10), another polar analogue bearing a carboxylate function, exhibited a K(i) value of 24 nM for A2B ARs, 49-fold selectivity versus human and 20-fold selectivity versus rat A1 ARs, and greater than 150-fold selectivity versus human A2A and A3 ARs. 8-[4-(2 Hydroxyethylamino)-2-oxoethoxy)phenyl]-1-propylxanthine (29) and 1-butyl-8-[4-(4 benzyl)piperazino-2-oxoethoxy)phenyl]xanthine (35) were among the most potent A2B antagonists showing K(i) values at A2B ARs of 1 nM, 57-fold (29) and 94-fold (35) selectivity versus human A1, ca. 30-fold selectivity versus rat A1, and greater than 400-fold selectivity versus human A2A and A3 ARs. The new potent, selective, water-soluble A2B antagonists may be useful research tools for investigating A2B receptor function. PMID- 11906293 TI - Novel 5-substituted 2,4-thiazolidinedione and 2,4-oxazolidinedione derivatives as insulin sensitizers with antidiabetic activities. AB - Two novel classes of 2,4-thiazolidinediones and 2,4-oxazolidinediones with an omega-(azolylalkoxyphenyl)alkyl substituent at the 5-position were prepared and their antidiabetic effects were evaluated in two genetically obese and diabetic animal models, KKA(y) mice and Wistar fatty rats. A large number of the 2,4 thia(oxa)zolidinediones showed potent glucose- and lipid-lowering activities. The antidiabetic activities of the 2,4-oxazolidinediones were superior to those of the 2,4-thiazolidinediones. Among the compounds, both enantiomers of 5-[3-[4-[2 (2-furyl)-5-methyl-4-oxazolylmethoxy]-3-methoxyphenyl]propyl]-2,4 oxazolidinedione (64), one of the most interesting compounds in terms of activity, were synthesized by using an asymmetric O-acetylation of the corresponding alpha-hydroxyvalerate (26) with immobilized lipase, followed by cyclization of the oxazolidinedione ring. (R)-(+)-64 showed more potent glucose lowering activity (effective dose (ED)25 = 0.561 mg/kg/d) than (S)-(-)-64 (ED25 > 1.5 mg/kg/d) or pioglitazone (ED25 = 6 mg/kg/d) in KKA(y) mice. It also exhibited a 10-fold more potent antidiabetic activity (ED25 = 0.05 mg/kg/d) than pioglitazone (ED25 = 0.5 mg/kg/d) in Wistar fatty rats. The antidiabetic effects of this compound are considered to be due to its potent agonistic activity for peroxisome proliferator-activated receptor gamma (EC(50) = 8.87 nM). PMID- 11906294 TI - Synthesis and biological evaluation of cycloalkylidene carboxylic acids as novel effectors of Ras/Raf interaction. AB - The protooncogenes Ras and Raf play important roles in signal transduction pathways regulated by mitogen-activated protein kinases. Mutations of Ras that arrest the protein in its active state are frequently implicated in tumor formation. We used Ras and Raf proteins in the yeast two-hybrid system to search for natural or synthesized substances capable of modulating Ras/Raf interaction by specifically binding to one of the interacting partners. We found that cycloalkylidene carboxylic acids enhanced Ras/Raf interaction by acting on the cysteine-rich domain of Raf. Several analogues of the active substance 2 cyclohexylidene propanoic acid were synthesized and the importance of the semicyclic double bond in the stabilization of Ras/Raf interaction was demonstrated. Variation of the size and the substituents of the cyclic system as well as the length of the carboxylic acid resulted in enhanced Ras/Raf interaction. PMID- 11906296 TI - Emergency treatment of chemical and thermal eye burns. AB - Chemical and thermal eye burns account for a small but significant fraction of ocular trauma. The speed at which initial irrigation of the eye begins, has the greatest influence on the prognosis and outcome of eye burns. Water is commonly recommended as an irrigation fluid. However, water is hypotonic to the corneal stroma. The osmolarity gradient causes an increased water influx into the cornea and the invasion of the corrosive substance into deeper corneal structures. We therefore recommend higher osmolarities for the initial rinsing to mobilize water and the dissolved corrosives out of the burnt tissue. Universal systems such as amphoteric solutions, which have an unspecific binding with bases and acids, provide a convenient solution for emergency neutralisation. Both conservative anti-inflammatory therapy and early surgical intervention are important to reduce the inflammatory response of the burnt tissue. In most severe eye burns, tenonplasty re-establishes the conjunctival surface and limbal vascularity and prevents anterior segment necrosis. PMID- 11906297 TI - Central corneal thickness, radius of the corneal curvature and intraocular pressure in normal subjects using non-contact techniques: Reykjavik Eye Study. AB - PURPOSE: To establish a population profile of central corneal thickness (CCT), radius of the corneal curvature (CC) and intraocular pressure (IOP) and the relationships between them using non-contact techniques. METHODS: We used a population-based random sample of 415 male and 510 female Caucasians aged 50 years and older. CCT and the radius of CC were measured with Scheimpflug anterior segment photography. IOP was measured with air-puff tonometry. RESULTS: The mean IOP of right eyes was 15.1 mmHg (SD 3.3) among men and 15.8 mmHg among women (SD 3.1), which is a statistically significant difference. The mean radius of CC for male right eyes was 7.78 (SD 0.60) and for females 7.62 (SD 0.58) which is also statistically significant. Mean CCT for male right eyes was 0.528 mm (SD 0.041) and for females 0.526 mm (SD 0.037), which is not a significant difference. Linear regression analysis shows no relationship between the radius of CC and IOP or between age and radius of CC. Linear regression analysis of the relationship between CCT and IOP suggests higher IOP measurements with thicker corneas. There was no significant correlation between age and CCT. CONCLUSION: IOP was found to be independent of age and significantly higher in females than in males. Radius of CC was found to be age-independent and significantly steeper in females than in males. CCT appears to be independent of age and gender. Greater CCT is associated with higher mean IOP. PMID- 11906298 TI - Results with a modified method for scleral suturing of intraocular lenses. AB - PURPOSE/METHODS: The long-term results obtained by a modified method of scleral suturing of posterior chamber lenses (PCLs) (Behndig & Otto 1997) were retrospectively registered and evaluated. Forty-four patients with insufficient capsular support for ordinary PCL implantation were included. Mean follow-up time was 13.1 +/- 6.4 months and registered parameters included visual acuity, deviation from planned refractive outcome, induced astigmatism, and complications. RESULTS/CONCLUSIONS: The results were comparable to those reported earlier for this type of surgery. It has previously been stated that scleral suturing of posterior chamber lenses is a safe procedure with good long-term results. This modified method produces comparable results while being more surgically simple and reproducible. PMID- 11906300 TI - Primary transpupillary thermotherapy (TTT) for malignant choroidal melanoma. AB - PURPOSE: To evaluate the efficacy of transpupillary thermotherapy (TTT) as the only method of treatment for small choroidal melanoma. PATIENTS AND METHODS: In a prospective non-randomized analysis, 20 patients with primary choroidal melanoma (posterior to the equator with base < or = 2 and thickness < or = 4.5 mm) were treated with TTT as the only method of treatment (diode laser at 810 nm, beam diameter 3 mm, power setting 0.3-0.9 W, exposure time 20-37 min). During follow up of at least 6 months, clinical aspects, ultrasonographic tumour thickness, fluorescein and indocyanine green angiographic patterns, visual acuity and ocular side-effects were recorded. RESULTS: In 17 eyes the tumour regressed significantly within 3 months after one treatment session (to a flat chorioretinal scar in 15 eyes). Despite a clinically flattened chorioretinal scar, fluorescein and indocyanine green angiography revealed that choriocapillary vessels in the heat-treated areas of 15 eyes remained perfused. Three amelanotic melanomas showed almost no response to TTT after repeated treatment at higher power settings. Visual acuity remained unchanged or improved in 12 eyes. Ocular side-effects included posterior synechia of the iris (1), macular oedema (2) and temporary retrobulbar pain (2). No patient showed tumour recurrence or metastases during follow-up. CONCLUSIONS: Preliminary results obtained by this study demonstrate good efficacy and visual outcome following TTT as the only method of treatment for small choroidal melanoma. However, indocyanine green angiographic findings suggest that tissue damage in the choroidal layer might be less effective, which perhaps may lead to a higher rate of tumour regrowth. Long-term follow-up is required to obtain data on late ocular side-effects, tumour recurrence and metastasis. PMID- 11906299 TI - Cataract surgery on diabetic patients. A prospective evaluation of risk factors and complications. AB - PURPOSE: This study presents an evaluation of cataract surgery on diabetic patients. One experienced surgeon carried out phaco emulsification on all subjects and the same surface-coated one-piece PMMA-lens-type was implanted. The lens fluorescence and the blood-aqueous barrier (BAB) were then evaluated as experimental preoperative risk indicators. RESULTS: During follow-up, 10 out of 39 diabetic patients progressed unilaterally in diabetic retinopathy or developed macular oedema, a significant relative risk. Neither lens fluorescence, BAB, HbA1c, level of retinopathy, type/duration of diabetes, diabetes treatment or antihypertensive treatment differed significantly between the group of patients with postoperative progression of retinopathy/macular oedema and those without. Results indicated NIDDM (non-insulin-dependent diabetes mellitus/type 2 diabetes) patients might have increased risk of a postoperative macular oedema. CONCLUSION: When diabetic retinopathy (DR) is not in a proliferative phase it should not be regarded as a contraindication to modern cataract surgery. Neither lens fluorescence nor BAB is valuable as a risk indicator for postoperative progression of DR. PMID- 11906301 TI - Histopathologically verified non-vascular optic nerve lesions in Denmark 1940-99. AB - PURPOSE: To evaluate the distribution in Denmark of histologically verified optic nerve lesions according to histological diagnosis, gender and age during a period of 60 years and to calculate frequency and possible changes in frequency during the observation period. METHODS: All optic nerve lesions reported at the Eye Pathology Institute, University of Copenhagen, during the period 1940-99 were investigated. All similar lesions reported at all pathological departments in Denmark during 1974-99 were also investigated. RESULTS: The number of optic nerve lesions identified totalled 313 in 298 patients. The frequency of histopathological optic nerve lesions was seen to have increased significantly over the last 25 years, and at present, an average of 12 lesions per year are recorded. Lesions in children represented 42% (130). The percentage of lesions suffered by children remained constant throughout the observation period. Benign tumours constituted 33% (44) of the total number of tumours in adults and 61% (62) in children. Frequencies of glioma and optic nerve sheath meningioma increased significantly during the last 25 years. Subjects presenting with optic nerve sheath meningioma exhibited a significant difference in age at presentation: the mean age of women at presentation was seen to be 48.8 years, while the mean age of men at presentation was seen to be 29.7 years. The most frequent lesion seen in children was glioma and in adults invasion from malignant uveal melanoma. CONCLUSION: Histologically diagnosed optic nerve lesions are rare and consist primarily of tumours. The increase in frequency of optic nerve lesions in Denmark during the last 25 years is due to an increase in the number of benign tumours. PMID- 11906302 TI - Genetic counseling in Leber hereditary optic neuropathy (LHON). AB - PURPOSE: To demonstrate the importance of mitochondrial DNA (mtDNA) analysis in the diagnosis of Leber hereditary optic neuropathy (LHON) and illustrate the difficulties in genetic counseling of the disease. PARTICIPANTS AND METHODS: Ophthalmological and molecular genetic study of one affected and three unaffected members from a family with heteroplasmic ND1/3460 mtDNA mutation associated with LHON. RESULTS: The proband had variable amounts of mutant mtDNA in all his tissues studied, ranging from 58% in blood to 92% in subcutis. The mother had an extremely low amount of mutant mtDNA in her tissues, except for hair roots, which contained only normal mtDNA. No mutant mtDNA could be detected in the proband's unaffected sister and maternal aunt. CONCLUSIONS: Despite her minimal mutation load, the mother of the proband has still transmitted a considerable amount of mutant mtDNA to her son, who is severely affected. Although proband's unaffected sister and maternal aunt had no mutant mtDNA, a theoretical risk that they may transmit the disease to their offspring cannot be excluded. PMID- 11906303 TI - The frequency of amblyopia among visually impaired persons. AB - PURPOSE: To investigate the frequency of amblyopia among visually handicapped patients. METHODS: The study is a retrospective investigation of all living patients registered in four Visual Rehabilitation Centres in a region in southern Sweden. The area's total population numbered 865,612 persons of whom 11,365 were registered as visually handicapped (with visual acuity < or = 0.3 in the better eye). RESULTS: Amblyopia was the main cause of decreased visual acuity in one eye in 1.72% (195 of 11,365) of the patients. The average age of the patients with amblyopia was 69 years (9-95 years) and 28.2% of these patients were less than 65 years old (the age for retirement in Sweden). The median visual acuity in the amblyopic eye among these patients was 0.1. The median visual acuity in the nonamblyopic eye was 0.2. The most common cause of decreased vision in the nonamblyopic eye was macular degeneration (39.5%). Bilateral amblyopia was present in 13 (6.7%) of the amblyopic patients. By comparing this study with earlier studies, we can calculate that about 1.2% of the persons with amblyopia 0.3 or lower will eventually become visually handicapped. CONCLUSION: A small but considerable number of patients who attend the Visual Rehabilitation Centres have amblyopia as a cause of their visual impairment. Since amblyopia can be treated if detected in childhood, later visual rehabilitation of these patients can be avoided or delayed, thereby reducing rehabilitation costs for society. PMID- 11906304 TI - Optic disc biomorphometry with the Heidelberg Retina Tomograph at different reference levels. AB - PURPOSE: The Heidelberg Retina Tomograph (HRT) is a confocal scanning laser tomograph that produces high resolution optical section images of the optic disc and central retina. Measurement accuracy and reproducibility is good. Several of the stereometric variables depend on the definition of a reference plane level. The purpose of the present study was to evaluate the four different reference levels in terms of their advantages and disadvantages in clinical work. METHODS: Sixty-seven randomly chosen eyes belonging to 67 subjects were included in this study. Forty of the eyes were healthy and 27 had glaucoma. The HRT with software versions 1.09 and 1.11 was used to acquire and evaluate topographic measurements of the optic disc. Image analysis was performed at four different reference levels: 320 microm fixed offset reference level (REFd) (version 1.09), an individually determined reference level (REFi), a papillo-macular reference level (REFm) and a flexible reference level (REFf) (version 1.11). ANOVA was used to determine differences in the topographic parameters between the reference levels. RESULTS: In terms of the healthy eyes, all the variables using different reference levels give rather similar results. However, with advanced glaucoma the measurement values provided with REFd are clearly different to those of the other reference levels. The measurement values using REFm and REFf provide fairly similar results in all clinical groups. REFf indicates the lowest point in the segment between 350 degrees and 356 degrees along the contour line and thus provides the most stable and clinically useful reference level at present. CONCLUSION: At present, the flexible reference level REFf gives the most reliable and adequate HRT measurement values, both in normal and in glaucomatous eyes. PMID- 11906305 TI - The short-term effect of latanoprost on intraocular pressure and pulsatile ocular blood flow. AB - PURPOSE: There is evidence that ocular blood flow plays a critical role in the clinical course of glaucoma. Any reduction in ocular blood flow due to topical antiglaucoma treatment should therefore be avoided. This study aimed to evaluate the short-term effect of local latanoprost application on ocular hemodynamics. METHODS: Intraocular pressure (IOP), ocular pulse amplitude (OPA), ocular pulse volume (OPV), systemic blood pressure, heart rate and the pulsatile component of ocular blood flow (POBF) were recorded using a pneumotonometer linked to the Langham Ocular Blood Flow System in 24 patients in a prospective, open-label study before and after 1 week of topical latanoprost application in both eyes. Twenty of the subjects had primary open-angle glaucoma and four had ocular hypertension. RESULTS: After 1 week of latanoprost treatment, IOP decreased significantly 6.2 +/- 2.9 mmHg in OD (P < 0.001) and 6.2 +/- 3.2 mmHg in OS (P < 0.001). Pulsatile OBF increased significantly by 201.2 +/- 167.4 microL/min in OD (P < 0.001) and 203.8 +/- 187.3 microL/min in OS (P < 0.001). Ocular pulse amplitude and OPV showed statistically significant increases (P < 0.05 and P < 0.001 respectively). Blood pressure and heart rate did not change significantly. CONCLUSION: Our results indicate that 1 week after latanoprost application, POBF, OPA and OPV were significantly increased in the eyes treated. More information on the perfusion of the optic nerve head is needed before the relevance of these findings to optic nerve head blood flow can be interpreted correctly. PMID- 11906306 TI - Visual functions and adverse ocular effects in patients with amiodarone medication. AB - PURPOSE: To study visual functions and ocular adverse effects of long-term amiodarone medication. METHODS: We performed an eye examination of 22 patients with long-term amiodarone medication. In addition to corrected visual acuity, colour vision was studied with the Standard Pseudoisochromatic Plates part 2 and Farnsworth-Munself 100 hue test. Contrast sensitivity was examined with the Pelli Robson chart. Visual fields were tested by Goldmann and Friedmann perimetry. RESULTS: Two patients with otherwise healthy eyes had abnormal blue colour vision test results. Otherwise colour vision, contrast sensitivity, and visual field test results were within normal range or could be explained by eye diseases such as cataract. Corneal drug deposits were found in 100% of the examined eyes. Slight anterior subcapsular lens opacities were found in 22.2%. Dry eyes were diagnosed in 9.1%. The eye fundi did not reveal any abnormalities that could be thought of as caused by amiodarone. CONCLUSION: The slight blue colour vision defect found in two patients with otherwise healthy eyes might represent an early sign of the optic nerve impairment which is a rare complication of amiodarone medication. The number of corneal and lens changes as well as dry eyes were found at levels previously described. PMID- 11906307 TI - Iris transluminance in type 2 diabetes. AB - PURPOSE: Cross-sectional analysis of iris transluminance in type 2 diabetic patients compared with control subjects, all followed in a cohort study. METHODS: A cohort consisting of a well-characterized group of 82 type 2 diabetic patients were followed for 10 years after diagnosis, as were 125 control subjects. The prevalence of iris transluminance was determined by transscleral transillumination and by grading of black-and-white positive prints at the 10 year examination. The frequency of diabetic retinopathy was prospectively determined by grading of fundus photographs at baseline and after five and 10 years. RESULTS: Iris transluminance was found in 27% of diabetic patients and 8.0% of control subjects (P = 0.001). The diabetic patients with more severe and short-term retinopathy more commonly had abnormal iris transluminance than those with no or only mild retinopathy (P < 0.05). Iris transluminance was not associated with intraocular pressure or glycemic control. CONCLUSIONS: In type 2 diabetic patients, abnormal iris transluminance tended to associate with more severe and short-term retinopathy. Therefore we assume that hypoxia is responsible for the defects in the pigment layer of the iris. Thus, abnormal iris transluminance may serve as a marker for rapidly progressive retinopathy. PMID- 11906308 TI - A clinicopathological study of venous loops and reduplications in diabetic retinopathy. AB - PURPOSE: To study possible causes of the gradual venous occlusion that precedes the formation of venous loops and reduplications in diabetic retinopathy using histopathological techniques. METHODS: Casts of the retinal vascular system from six eyes of five diabetic patients were used to identify venous loops and reduplications. Subsequently the lesions were studied by immunohistochemistry using antibodies against cellular and non-cellular components previously shown to be disturbed in the diabetic retina. RESULTS: In all eyes the venous abnormalities identified on the casts were accompanied by abnormal wall partitions dividing the venous lumen. These partitions consisted of a double layer of flat cells displaying immunoreactivity to von Willebrand factor and actin (endothelial cells), but not to glial fibrillary acid protein or S-100 protein (glial cells), CD3, CD20, CD68, or neutrophil elastase (leucocytes). Neutrophile granulocytes adhering to the walls of larger retinal venules were unrelated to the venous partition and to capillary occlusion in the adjacent retinal areas. CONCLUSIONS: Venous loops and reduplications are associated with partitions of the larger retinal venules consisting of a double layer of endothelial cells anchored to a thin basement membrane. An elucidation of the factors distinguishing this endothelial cell proliferation from preretinal new vessel formation may be important for understanding the pathophysiology of proliferative diabetic retinopathy. PMID- 11906309 TI - Transplantation of allogenic anterior lens capsule to the subretinal space in pigs. AB - PURPOSE: To investigate the consequences of transplantation of a new basement membrane to the subretinal space (SRS) as a substitution of Bruch's membrane. METHODS: Porcine anterior lens capsules (ALC) were transplanted to the subretinal space of 20 eyes from 19 young Danish landrace pigs. All pigs underwent a three port localized pars plana vitrectomy. Seventeen eyes received naked ALC. In three experiments the ALC was embedded in gelatine, in order to prevent curling of the ALC. The observation period varied between zero and 49 days. The pigs were examined by ophthalmoscopy and fundus photography. Histopathological examination of enucleated eyes was performed at the end of the experiment. RESULTS: ALCs transplanted to the subretinal space were well-tolerated and caused no inflammation when Bruch's membrane was left undamaged. After 11 days host RPE and glial cells started to cover the ALC in a competitive fashion. When Bruch's membrane was damaged, ingrowths of choroidal vessels and fibroblasts was prominent. The use of gelatine to flatten the ALC did not prevent curling, and gelatine caused pronounced inflammation. CONCLUSIONS: It is possible to transplant porcine ALC to the SRS of the pig. ALCs are well-tolerated in the SRS and are covered with well-differentiated monolayers of host RPE-cells, if Bruch's membrane is left intact. PMID- 11906310 TI - Optical coherence tomography of choroidal neovascularization in high myopia. AB - AIM: To investigate morphologic changes in the eye with choroidal neovascularization (CNV) in high myopia using optical coherence tomography (OCT). METHODS: Optical coherence tomography was performed in 35 patients (42 eyes) with myopic CNV. Myopic CNV was divided into active, scar, or atrophic stages based on funduscopic and fluorescein angiographic findings. The characteristics of OCT findings in each stage were identified. RESULTS: In the active stage (11 eyes), OCT clearly displayed a neovascular membrane as a highly reflective dome-like elevation above the retinal pigment epithelium (RPE). No apparent subretinal fluid accumulation around the CNV was identified. In the scar stage (12 eyes), only the surface of the CNV showed high reflectivity, which was markedly attenuated below the surface. In the atrophic stage (19 eyes), the CNV had become totally flat and chorioretinal atrophy around the regressed CNV showed high reflectivity. CONCLUSIONS: Optical coherence tomography demonstrated characteristic features at each stage of myopic CNV. Optical coherence tomography appears to be useful in evaluating the stage and activity of myopic CNV. PMID- 11906311 TI - Vitreous surgery combined with internal limiting membrane peeling for traumatic macular hole with severe retinal folds. AB - PURPOSE: To report a case of a traumatic macular hole with severe retinal folds in which vitreous surgery combined with internal limiting membrane (ILM) peeling was beneficial. To demonstrate that the area from which the ILM was peeled can be clearly differentiated by scanning laser ophthalmoscopy. METHODS: A posterior hyaloid detachment was created during vitreous surgery on a 34 year old man with a traumatic macular hole. The remaining vitreous was resected and the ILM was peeled. The fundus was examined through a scanning laser ophthalmoscope before and after the surgery. RESULTS: The retinal folds disappeared concurrently with the detachment of the ILM, resulting in closure of the macular hole. The area from which the ILM was peeled was clearly observed through the scanning laser ophthalmoscope. CONCLUSION: In this patient, it was helpful to perform not only posterior hyaloid detachment but also ILM peeling. The scanning laser ophthalmoscope was highly useful for observing the area from which the ILM was peeled. PMID- 11906312 TI - Spontaneous remodelling of retinal veins accidentally severed during vitreous surgery. AB - BACKGROUND: To report the spontaneous remodelling of retinal veins inadvertently severed during vitreous surgery. METHODS: Four diabetic patients who underwent vitrectomy during which retinal veins were damaged were followed for at least 6 months. Haemorrhages from severed veins were controlled by raising intraocular pressure and by endodiathermy of the distal ends of the veins. This was followed by fluid/gas exchange. RESULTS: Remodelling of venous channels in the damaged areas was observed in all eyes. In three of the four eyes we found evidence of collateral veins. In the other eye we noted the connection of two ends of an interrupted vein. No signs of retinal vein occlusion (e.g. superficial retinal haemorrhages and retinal oedema) were observed. CONCLUSION: These findings suggest that a spontaneous remodelling mechanism to repair damaged venous channels may exist in retina. PMID- 11906313 TI - A case of Coats' disease with a peeling of premacular fibrosis after photocoagulation. AB - We report a 26-year-old man with Coats' disease associated with premacular fibrosis. As an initial treatment, the peripheral exduative area was treated with argon laser photocoagulation. Six weeks later, the premacular fibrosis was peeled off and the posterior vitreous membrane was also detached. The patient's visual acuity improved to 20/20. We also observed a change of the vitreous component before and after the treatment that was similar to posterior vitreous detachment (PVD). This is the first reported case in which a distinct vitreous change was observed after premacular fibrosis peeling in Coat's disease. PMID- 11906314 TI - Bilateral Coats' disease: long-term follow up. AB - PURPOSE: To report on the long-term follow-up of a female patient with bilateral Coats' disease, who showed marked asymmetry between the two eyes. METHODS: A five year old girl presented in 1978 with leukocoria in a blind right eye. A total exudative retinal detachment and extensive retinal telangiectasiae were noted. In the other eye, there was a localized area of retinal exudation and vascular abnormality in the supero-temporal periphery. Ultrasonography showed no evidence of intraocular tumour in the right eye and a clinical diagnosis of bilateral Coats' disease was made. RESULTS: In 1995, the area or retinal exudation in the left eye increased and laser photocoagulation was applied successfully. To date, no disease recurrences have occurred. CONCLUSION: Although Coats' disease is usually unilateral, bilateral, asymmetrical involvement may occur on rare occasions. Long-term follow-up of the least affected eye is necessary so that late complications can be identified early and treated adequately to prevent visual loss. PMID- 11906315 TI - Intravitreal sustained-release ganciclovir implants for severe bilateral cytomegalovirus retinitis after stem cell transplantation. AB - PURPOSE: To describe the treatment of cytomegalovirus (CMV) retinitis with intravitreal sustain-release ganciclovir devices in a 16-year-old patient in third remission of acute lymphoblastic leukemia after stem cell transplantation. METHODS: The patient received a stem cell transplant from an unrelated bone marrow donor after which he contracted a serious CMV infection manifested in the lungs and retinae. His immune system at this time was almost completely depleted. Implantation of a sustained-release ganciclovir device was performed in both eyes when retinitis progressed in spite of aggressive antiviral intravenous treatment. RESULTS: No per- or postoperative complications were noted. Infiltrates, hemorrhages and macular edema present preoperatively dissolved over a period of six months. The final visual acuity was 1.0 in both eyes. The patients immune system and lung function slowly recovered during the same time period. CONCLUSIONS: The intravitreal ganciclovir implant provides safe and effective therapy against CMV retinitis, and should be considered in patients acquiring the infection after stem cell transplantation. PMID- 11906316 TI - Survival of transplanted human corneal stem cells. Case report. AB - PURPOSE: To report a case history involving long-term survival of transplanted human corneal stem cells. METHODS: A male patient with severe bilateral chemical burns received six corneal transplants, all of which failed. He subsequently received combined corneal transplants and stem cell transplants, which have remained clear for 3 and 4 years respectively. One of the donors was female. We studied the gender of the epithelial cells of the cheek of the patient and of the two grafts using fixation and fluorescent in situ hybridization (FISH) analyses. RESULTS: In the graft from the female donor, 30% of the epithelial cells were of female origin. All the epithelial cells from the cheek and the other graft were of male origin. CONCLUSION: This case demonstrates that transplanted human corneal stem cells can survive and replicate in the long-term (3 years) without systemic immunosuppression. The case also indicates that a minority (30%) of healthy transplanted epithelial cells is enough to present a clear graft with a clinically healthy ocular surface. PMID- 11906317 TI - Acquired anomalous head posture following loss of vision in one eye. AB - BACKGROUND: We studied the anomalous head postures (AHPs) of five monocular viewing patients and investigated the possible causes and the appropriate surgical strategies to correct each condition. METHODS: Five patients with acquired visual loss in one eye and associated head tilt and/or turn were examined and treated for correcting the head posture according to the etiology of their respective AHPs. RESULTS: Three types of anomalous head position have been detected: head tilt related to cyclotropia, face turn associated with adduction blocked monocular nystagmus, and face turn to centre the visual field. Surgical plans were prepared according to the mechanism of the AHP in question. After surgery, all patients showed a marked reduction of the head tilt, except one who had a recurrence of the face turn 1 week postoperatively. Mean follow-up time was 19 months. Horizontal transposition of the vertical muscles for correcting cyclotropia offered stable normalization of the AHP in three monocular viewing patients with head tilt, and represents a safe, viable and easy alternative to the Harada Ito procedure. Horizontal recession of the medial rectus of the fixing eye minimized the abduction nystagmus and relieved the need to adduct the fixing eye and subsequently rotate the head toward the fixing eye in one patient. Recurrence of the AHP occurred in one patient. CONCLUSION: Different mechanisms may account for AHP in monocular viewing patients. Different surgical procedures may be used to correct the anomalous position. Careful patient selection and etiological diagnosis of AHP is required prior to developing a surgical strategy. PMID- 11906318 TI - Orbital metastasis as the initial manifestation of small cell lung cancer. PMID- 11906319 TI - Dysregulation of angiotensin II synthesis is associated with salt sensitivity in the spontaneous hypertensive rat. AB - (1) Salt sensitive hypertension, which occurs as a result of treatment with nitric oxide synthase inhibitors, is associated with a loss of the usual down regulatory effect of dietary sodium on angiotensin II (Ang II) synthesis. In the spontaneous hypertensive rat (SHR), which suffers a relative NO deficiency, the hypertension is in part salt sensitive. We sought to determine therefore whether the salt sensitive component to the hypertension was associated with a loss of the regulatory effect of dietary sodium on Ang II synthesis. (2) Male SHR were placed on low, intermediate or high salt diets for 4 weeks and their blood pressure recorded. After 4 weeks, blood was collected for determination of renin, angiotensinogen, Ang I, Ang II and aldosterone concentrations, as well as ACE activity. (3) The increase in systolic blood pressure in rats on the high salt diet was significantly greater than in those on the low (P < 0.005) and intermediate salt diets (P < 0.0005). Plasma renin and aldosterone concentrations and ACE activity decreased with increasing dietary sodium. However, the concentrations of Ang II and angiotensinogen both increased in the rats on the high salt diet (Ang II: P < 0.05; angiotensinogen: P < 0.05). (4) We conclude that the hypertension in the SHR is in part salt sensitive and that this salt sensitive component is associated with a loss of the normal down-regulatory effect of dietary sodium on Ang II and angiotensinogen synthesis. PMID- 11906320 TI - Effects of nitric oxide donors and inhibitors of nitric oxide signalling on endothelin- and serotonin-induced contractions in human placental arteries. AB - In order to explore the role of nitric oxide (NO) in the control of fetoplacental vascular tone in normal pregnancy we have examined the effects of NO donors on uteroplacental arteries pre-contracted with the vasoconstrictor endothelin-1 (ET 1) or serotonin (5-HT). We have furthermore examined the effects of guanylate cyclase inhibitors on the NO-induced relaxation. Segments of placental arteries (n=102) obtained from 39 placentas immediately after delivery were mounted in organ baths and superfused with Krebs-Ringer solution at 37 degrees C. The vessel segments were exposed to drugs for various intervals and the tension was recorded isometrically and registered on a polygraph. Cyclic guanosine monophosphate (cGMP) analysis was performed after extraction of vessel segments using a specific radioimmunoassay. The placental artery segments responded to ET-1 and 5 HT with a dose-dependent vasoconstriction. After pre-contraction with ET-1 (10( 7) M) or 5-HT (10(-6) M), the vessels relaxed in response to the NO donors glyceryltrinitrate (GTN) (10(-6) M) and S-nitroso-N-acetyl-penicillamine (SNAP) (10(-5) M). In the presence of the non-specific guanylate cyclase inhibitor LY 83583 (10(-6) M), the vessels responded with a small contraction. In the presence of the soluble guanylate cyclase (sGC) inhibitor 1H[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (ODQ) the non-treated vessels responded with a relaxation. 1H[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one gave no obvious relaxation in pre contracted vessels. Addition of 8-Br-cGMP, the cell-permeant analogue of cGMP, with or without pre-contraction had no effect on the vessels. Cyclic guanosine monophosphate analysis showed that GTN treatment caused an increase in cGMP after 12 min. Our results indicate that NO acts as a vasodilator in placental vessels. The cGMP-dependent mechanisms may be involved in NO-induced relaxation but cGMP independent mechanisms appear also to be involved. PMID- 11906321 TI - Erythropoietin concentrations during 10 days of normobaric hypoxia under controlled environmental circumstances. AB - Serum erythropoietin levels (s-[epo]), haemoglobin concentration ([Hb]), haematocrit (hct), and ferritin concentration ([fer]) were measured in seven healthy male volunteers (20-23 years) exposed continuously to hypoxia (PO(2) 14 kPa) for 10 days. Serum erythropoietin concentration increased significantly from 9.5 +/- 3.51 to 33.6 +/- 11.64 U L(-1) (P < 0.05) after 2 days of hypoxia. Thereafter, s-[epo] decreased. However, after 10 days s-[epo] was 18.7 +/- 5.83 U L(-1) which was still increased above the pre-hypoxia level (P < 0.05). Serum haemoglobin concentration and hct increased over the 10 days of hypoxia, [Hb] from 152 +/- 8.9 to 168 +/- 9.2 gL(-1) (P < 0.001), and hct from 43 +/- 2.4 to 49 +/- 2.6% (P < 0.001). Ferritin concentration decreased significantly during the hypoxic exposure from 82 +/- 46.9 to 44 +/- 31.7 mmol L(-1) after 10 days (P < 0.01). In conclusion, the initial increase of s-[epo] under controlled normobaric hypoxia was marked, 353%, and levelled off after 5-10 days at 62-97% above normoxia level. There was also a significant increase in [Hb] and hct and a decrease in [fer] after 10 days of exposure to normobaric hypoxia. PMID- 11906322 TI - Influence of controlled hypoxia and radical scavenging agents on erythropoietin and malondialdehyde concentrations in humans. AB - The present study was carried out to evaluate a model of a hypoxic stimulus of erythropoietin (EPO) production in humans and to investigate the role of free oxygen radicals in human EPO production. The study was conducted as an open, randomized, parallel, placebo-controlled trial. Thirty-six healthy male volunteers received a hypoxic treatment (13% O(2)) with a respiration mask for 6 h. During the period of hypoxia, the volunteers received as a short-term treatment either 1200 mg thioctic acid, or N-acetylcysteine 600 mg or placebo (0.9% sodium chloride). The EPO concentration in plasma increased up to 290% of the baseline level in all three groups. No statistically significant differences of AUC(EPO(0-48 h)) could be demonstrated between the groups. The malondialdehyde (MDA) concentration in plasma increased significantly (P < 0.001) 2 h after termination of hypoxia (mean 129.8 +/- 6.8% of the baseline) in all three groups. CONCLUSIONS: Taken together, our in-vivo results do not support a gross modulatory effect of a short-term treatment with radical scavenging agents on EPO production during or after hypoxia in humans, as derived from the detected changes of MDA-concentrations in peripheral plasma. PMID- 11906323 TI - PKC-alpha shows variable patterns of translocation in response to different stimulatory agents. AB - Reports from numerous laboratories suggest that protein kinase C (PKC) translocation to substrate target sites may vary depending on cell type and experimental conditions. We have proposed that acutely variable targeting of PKC to different substrate sites could greatly expand the functional properties of individual isoforms in individual cell types (Li et al., 2001). Confocal microscopy and PKC alpha-enhanced green fluorescent protein (PKC alpha-EGFP) fusion protein expression were utilized to investigate the spatial and temporal pattern of PKC alpha translocation to different stimulating agents in A7r5 smooth muscle cells. Phorbol 12, 13 dibutyrate (PDBu 10(-8) M) caused a slow but irreversible relocation of the fusion protein from the cytosol to the plasmalemma. By comparison, thapsigargin (10(-5) M) and A23 187 (2 x 10(-5) M) induced a rapidly transient translocation to the cell membrane which was completed within 4 min. In contrast to these agents, angiotensin II (Ang II, 10( 6) M) caused only partial relocalization of cytosolic PKC alpha-EGFP to brightly fluorescing patches at the cell periphery. Localization at peripheral patches was completed within seconds and the fusion protein returned to the cytosol within 2 min. The PKC inhibitor staurosporine blocked cellular contraction to PDBu but not A(23 187) and had no effect on PKC alpha-EGFP translocation. By comparison, the calcium chelators EDTA and BAPTA-AM blocked the contraction to A(23 187), attenuated the contraction to PDBu, and abolished the translocation of PKC alpha EGFP by both agents. The results show that in a single cell type the spatial and temporal characteristics of individual PKC isoform translocation may differ markedly. This further suggests the existence of potentially complex mechanisms which regulate the rate and location of target site availability. PMID- 11906324 TI - Immunomodulatory action of chronic exercise on macrophage and lymphocyte cytokine production in mice. AB - This study was designed to evaluate the effects of 8-week voluntary running exercise on cytokine production of macrophages and lymphocytes. Seven-week-old male BALB/c inbred mice were divided into two groups: a group given voluntary exercise (exercise group, n=32), and the other, a non-exercise group (control group, n=32). Exercise consisted of spontaneous running in wheels for 3 days per week over 8 weeks. The levels of nitric oxide (NO2-) and interleukin (IL)-1 beta production by lipopolysaccharide (LPS)-stimulated peritoneal macrophages from the exercise group was significantly higher than that in the control group (P < 0.05 P < 0.01). In the exercise group, stimulation indices by concanavalin A (Con A) was significantly higher than they were in the control group (P < 0.05-P < 0.001). When compared with the control group, the exercise group showed a significant (P < 0.05) increase in the splenic lymphocyte production of IL-2 stimulated by Con A (449.5 +/- 28.2 and 853.7 +/- 116.0 pg 4 x 10(5) cells(-1) 48 h(-1) for the control group and the exercise group, respectively). IL-4 production of splenocytes stimulated by Con A in the exercise group (37.6 +/- 5.1 pg 4 x 10(5) cells(-1) 48 h(-1)) was higher than that in the control group (30.9 +/- 3.9 pg 4 x 10(5) cells(-1) 48 h(-1)); however, the difference was not statistically significant. These results suggest that 8-week voluntary running exercise effectively enhanced macrophage and lymphocyte functions in mice. PMID- 11906325 TI - Serosal application of Ba(2+) induces oscillatory chloride secretion via activation of submucosal cholinergic neurones in guinea-pig distal colon. AB - The enteric nervous system regulates ion and fluid secretion in the mammalian intestine at both resting and stimulated conditions. To determine the type and activation mechanism of neurones involved, mucosa-submucosa sheets isolated from guinea-pig distal colon were studied in vitro in Ussing chambers. Serosal addition of 0.5-1 mM barium (Ba(2+)), a potassium (K(+)) channel inhibitor, caused oscillatory increases in short-circuit current (I(sc)). Mean values of the size and frequency of I(sc) were 369.1 microA cm(-2) and 2.3 min(-1). The oscillatory I(sc) induced by the low concentrations of Ba(2+) was blocked by either higher concentrations of Ba(2+) (2-5 mM) or other K(+) channel inhibitors, such as tetraethylammonium (TEA) (1 mM) and quinine (20 mM). The Ba(2+)-induced oscillatory I(sc) was also inhibited by tetrodotoxin (TTX) and atropine. In a nominally Ca(2+) free solution plus serosal addition of 0.1 mM ethylene glycol bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), a Ca(2+) chelator, the oscillatory I(sc) slowed and diminished. Further, the Ba(2+)-induced oscillatory I(sc) was partially inhibited by apical addition of 100 microM 5' nitro-2-(3-phenylpropylamino)benzoic-acid (NPPB), a Cl(-) channel inhibitor, and completely disappeared in a low Cl(-) solution (11 mM) on both sides. On the other hand, application of either cimetidine, a histamine H(2) receptor antagonist, or hexamethonium, a nicotinic antagonist, to the serosal side did not affect the Ba(2+)-induced oscillatory I(sc). In conclusion, the Ba(2+)-induced oscillatory I(sc) is the transepithelial Cl(-) current which is stimulated by activation of cholinergic neurones in submucosal plexus of guinea-pig distal colon. PMID- 11906326 TI - Expression of cyclooxygenase-2 in intestinal goblet cells of pre-diabetic NOD mice. AB - Cyclooxygenase, the rate-limiting enzyme in prostaglandin synthesis, is expressed in constitutive (COX-1) and inducible (COX-2) isoforms. The COX-2 has been proposed to be involved in development of autoimmune type 1 diabetes (T1D). We examined COX-2 expression in the gut-associated lymphoid tissue (GALT), and found COX-2 was strongly expressed in goblet cells of non-obese diabetic (NOD) mice at the apical villi at the age of 2.5 weeks, clearly before the onset of insulitis, while the expression in the control BALB/c mice was weak or absent at all ages (P < 0.001). Lipopolysaccharide (LPS) given intraperitoneally slightly increased COX 2 expression in the goblet cells and epithelium of both NOD and BALB/c mice. High resolution confocal microscopy showed that the surroundings of the goblet cells contained no COX-2, implying that the enzyme is synthesized by the goblet cells. The COX-2 is secreted from goblet cells into the intestinal lumen along with mucins. The COX-2 concentration in the goblet cell of BALB/c and especially of NOD mice was markedly higher than that in the intraepithelial lymphocytes or lamina propria macrophages. High mucin COX-2 from goblet cells may increase luminal prostaglandin synthesis, alter epithelial permeability, modulate intestinal immune responses and modify functional properties of the lymphocytes in the GALT, which all may be important for the initiation of the autoimmune phenomenon in the NOD mice. PMID- 11906327 TI - Effect of exercise on concentrations of free amino acids in pools of type I and type II fibres in human muscle with reduced glycogen stores. AB - A few animal studies have shown that some amino acid concentrations vary between different muscle fibre types. In the present study, amino acid concentrations were measured in separate pools of different fibre types in human skeletal muscle, with reduced glycogen stores, before and after sustained exercise. Five subjects exercised at a submaximal work rate for 60 min and then at a maximal rate for 20 min. Biopsy samples were taken from the vastus lateralis muscle before and after exercise; they were freeze-dried and individual fibres were dissected out. Fragments of these fibres were stained for myosin-adenosine triphosphatase (ATPase) and identified as type I or type II fibres. The concentrations of free amino acids were measured by high performance liquid chromatography (HPLC) in perchloric acid (PCA) extracts containing pools of either type of fibre. After exercise, glycogen was decreased in type I fibres (53%) and in four subjects also in type II fibres. The concentrations of most amino acids were similar in the two fibre types before exercise, but the glutamate, aspartate and arginine levels were 10% higher in type II than in type I fibres. After exercise, the glutamate concentration was decreased by 45% in both fibre types and the branched-chain amino acids (BCAA) were decreased in type II fibres (14%). Exercise caused an increase by 25-30% in tyrosine concentration in both type I and type II fibres. The results show that amino acids can be measured in pools of fibre fragments and suggest that amino acid metabolism play an important role in both type I and type II fibres during exercise. PMID- 11906328 TI - Effect of endurance training on oestrogen receptor alpha transcripts in rat skeletal muscle. AB - Endurance training induces, in female rats, alterations of oestrous cycle with decrease in plasma oestradiol levels. Moreover, it is well known that oestradiol concentrations modify oestrogen receptor levels. In order to further explain the effects of oestrogens on skeletal muscles, we hypothesized that endurance training modifies the levels of oestrogen receptor alpha messenger ribonucleic acid (ER alpha mRNA) in rat gastrocnemius muscle. Wistar rats were separated into four groups: male controls (C(m)) (n=7), female controls (C(f)) (n=6), male trained (E(m)) (n=7) and female trained (E(f)) (n=6). The endurance training programme was performed for 7 weeks, 5 days week-1 and consisted of 1 h of continuous running on an adapted motor-driven treadmill. At the end of the training session, the gastrocnemius muscle was isolated, weighed and semiquantification of ER alpha mRNA was performed using the reverse transcriptase polymerase chain reaction (RT-PCR) technique. The citrate synthase (CS) activity of the gastrocnemius muscle was measured by a fluorimetric method. The CS activity of the male and female gastrocnemius muscle, respectively, 100 +/- 7% in C(m) (n=7) vs. 120 +/- 14% in E(m) (n=6, P < 0.01) and 100 +/- 13% in C(f) (n=6) vs. 138 +/- 23% in E(f) (n=6, P < 0.01) was significantly increased after 7 weeks of training. The ER alpha mRNA levels were significantly increased in E(f) compared with C(f) (0.49 +/- 0.15 vs. 0.31 +/- 0.11, P < 0.01) but not in E(m) compared with C(m) (0.37 +/- 0.15 vs. 0.37 +/- 0.13). In conclusion, these results demonstrate that 7 weeks of endurance training increased the level of transcripts encoding ER alpha in rats with the increase restricted to the females. PMID- 11906329 TI - Renal denervation abolishes the protective effects of ischaemic preconditioning on function and haemodynamics in ischaemia-reperfused rat kidneys. AB - Studies were conducted to investigate the role of renal sympathetic nerves in the process of acquiring ischaemic tolerance in ischaemic preconditioned ischaemia reperfused rat kidneys. Two periods of 3-min occlusion of bilateral renal arteries was performed prior to 30-min bilateral ischaemia and 90-min reperfusion in acute renal denervated or innervated kidneys. The glomerular filtration rate (GFR), fractional excretion of sodium (FENa) and lithium (FELi), and renal blood flow (RBF) were assessed in reperfused kidneys. Ischaemic preconditioning significantly improved values for all these parameters as compared with no treated ischaemia-reperfused kidneys. Denervation caused slight increase in GFR, diuresis and natriuresis without improving RBF after reperfusion. However, protecting effects of ischaemic preconditioning on renal function were disappeared in denervated kidneys, while in innervated kidneys the effects of ischaemic preconditioning were maintained. These results clearly showed that ischaemic preconditioning pre-treatment protects kidneys against ischaemia reperfusion injury, and the effects are, at least in part, mediated by sympathetic nerves, as the protective effects were abolished by denervation. PMID- 11906330 TI - Probiotics against allergy: data, doubts, and perspectives. PMID- 11906331 TI - CD14: an example of gene by environment interaction in allergic disease. PMID- 11906332 TI - Characterization of specific IgE response in vitro against protein and drug allergens using atopic and normal donors. AB - BACKGROUND: As the incidence of allergy to different compounds increases in society, the need to understand and characterize specific IgE responses becomes obvious. Different cell culture systems have been evaluated for their ability to support such IgE secretion. METHODS: One system employed human peripheral lymphocytes (PBL) from normal donors stimulated with anti-CD3 activated T cells with or without the presence of allergens like benzylpenicillin (BP) and Phlenum pratense (PP). Secretion of IgE was analyzed in ELISA and compared to the IgG response to the nonallergenic antigen tetanus toxoid (TT). Another system employed stimulation of T and B cells with a heterotope, consisting of a T helper cell epitope derived from TT, and a B cell allergen epitope derived from BP. The specific IgE secretion was compared, using lymphocytes from normal as well as BP allergic donors. RESULTS: Anti-CD3 stimulated T cells supported BP-specific IgE secretion in six of 11 normal donors. This response was inhibited in four donors and enhanced in two donors by the addition of the BP-allergen to the culture. In contrast, addition of the protein allergen (PP) or antigen (TT) to the same culture system inhibited both IgE and IgG synthesis in all experiments. Cells from the majority (10/16) of the BP-allergic donors failed to produce BP-specific IgE in vitro, when cultured in the presence of allergen. CONCLUSIONS: An allergen specific immune response is readily generated in vitro. The differential response against benzylpenicillin between different donor categories most probably reflects the level of pre-exposure to this allergen in vivo. PMID- 11906333 TI - Can the standard gamble and rating scale be used to measure quality of life in rhinoconjunctivitis? Comparison with the RQLQ and SF-36. AB - BACKGROUND: With interest in health economics growing, it is important to know whether utilities may be used to measure health-related quality of life in patients with rhinoconjunctivitis. The objective was to compare the validity and measurement properties of disease-specific versions of the standard gamble and rating scale with those of the Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) and the Short-Form 36 (SF-36). METHODS: One hundred adults with symptomatic rhinoconjunctivitis participated in a 5 week observational study, completing the standard gamble, rating scale, RQLQ and SF-36 at baseline and after 1 and 5 weeks. Symptom diaries were completed for 1 week before each follow up visit. RESULTS: Reliability was highest for the RQLQ (intraclass correlation coefficient = 0.97), followed by the rating scale (0.75), the SF-36 physical (0.75), the SF-36 mental (0.74) and the standard gamble (0.12). The responsiveness index was highest for the RQLQ (0.76), followed by the rating scale (0.56) and the SF-36 mental (0.28). Both cross-sectional and longitudinal validity were strongest for the RQLQ and the rating scale. CONCLUSIONS: Both the rating scale and the RQLQ have strong evaluative and discriminative properties. The SF-36 has acceptable discriminative properties but its evaluative properties are poor. All measurement properties for the standard gamble are inadequate. Poor correlation between the standard gamble and the rating scale indicates that utilities cannot be derived from rating scale data. PMID- 11906334 TI - Cellular immune responses to ovalbumin and house dust mite in egg-allergic children. AB - BACKGROUND: Although IgE-mediated food (egg) allergy is typically lost with age the underlying immune mechanisms are not understood, particularly in relation to the development of persistent IgE-mediated aeroallergen sensitivity. METHODS: Lymphoproliferation and cytokine responses (IL-5, IL-10, IL-13 and IFN-gamma) to house dust mite (HDM) allergen and egg ovalbumin (OVA) were assessed using peripheral blood mononuclear cells (PBMC) from children aged 6 months to 5 years (n = 59) with acute IgE-mediated egg allergy (urticaria and angiedema or anaphylaxis), as confirmed by positive skin prick testing (SPT). Of these 46 had positive SPT on the day of blood collection and 13 had outgrown egg allergy (negative SPT and successful egg challenge). Where possible, responses were compared with previous data from nonallergic children of similar ages (n = 107). RESULTS: Transient lymphoproliferative responses to OVA were seen in both egg allergic and nonallergic children, but were more marked and more prolonged in egg allergic children. Younger egg-allergic children (< 18 months) showed a mixed Th0 cytokine response to OVA, with readily detectable IFN-gamma, IL-5, IL-13 IL-10. Although IL-13 and IL-5 responses (OVA) correlated in younger egg-allergic children, there was a dissociation of these Th2 responses with age. Loss of clinical reactivity to egg was associated with almost complete loss of IL-5 responses and OVA-specific lymphoproliferation. Although IL-13 levels tended to be lower with age, this was not significant. Strong IFN-gamma and IL-10 responses to OVA persisted in older children after loss of OVA-specific lymphoproliferation. Lymphoproliferative responses to HDM also developed earlier in egg-allergic children compared with nonallergic children. Th1 (IFN-gamma) responses to HDM were largely below detection prior to 18 months of age, but increased significantly with age. In egg-allergic children Th2 (IL-5, IL-13) HDM responses also progressively increased with age. At 3 years of age almost all egg allergic children had positive SPT to HDM and positive lymphoproliferative responses to HDM, with strong Th1 and Th2 (Th0) cytokine production. CONCLUSIONS: IL-5 responses (rather than IL-13) responses most closely reflected clinical food allergy, with dissociation of IL-5 and IL-13 responses in older and egg-tolerant children. In this population, food and aeroallergen sensitivity was not associated with inability to produce IFN-gamma, but rather with mixed Th2 and Th1 (Th0) responses. Strong IL-10 and IFN-gamma responses where associated with the development of tolerance, suggesting persistent 'regulatory' populations of OVA specific T cells, rather than clonal deletion of OVA responsive T-cells. PMID- 11906335 TI - Molecular cloning and sequence analysis of full-length cDNAs encoding new group of Cyn d 1 isoallergens. AB - BACKGROUND: Cyn d 1, the major allergen of Bermuda grass pollen, contains some acidic/basic isoforms. The N-terminal amino acid sequences of some acidic Cyn d 1 isoforms were found to be different from those of Cyn d 1 cDNA clones identified previously. METHODS: A predicted 17-meric oligonucleotide probe was designed to fish the unidentified isoallergen cDNAs out of BGP cDNA library. The reactive clones were isolated and verified by sequencing. Two of them were expressed in the yeast Pichia pastoris to obtain recombinant Cyn d 1 proteins. RESULTS: All four cDNA clones encode the full-length Cyn d 1 with mature proteins of 244 amino acid residues. A 97-99% identity was found among the deduced amino acids of these four clones while an 86% identity was elicited between the four clones and the ones previously identified. The predicted isoelectric focusing (pI) values of the newly identified Cyn d 1s are acidic while pIs of the previously identified Cyn d 1s are basic. The two recombinant acidic Cyn d 1 proteins possess the epitopes recognized by mouse and rabbit polyclonal anti-Cyn d 1 antibodies, and have human IgE-binding capacity as revealed by immunodot assay. CONCLUSIONS: The present study identified full-length cDNAs encoding new isoallergens of Cyn d 1, and separated Cyn d 1 gene into an acidic group and a basic group. PMID- 11906336 TI - Purification and characterization of Pla a 1, a major allergen from Platanus acerifolia pollen. AB - BACKGROUND: Plane trees, as Platanus acerifolia, are an important source of airborne allergens in cities of the United States and Western Europe. Little is known about the relevant allergens of this pollen. The aim of this study was to identify relevant allergens from P. acerifolia pollen and purify and characterize a major allergen of 18 kDa. METHODS: P. acerifolia pollen extract was fractionated using ion exchange, gel filtration, and reverse-phase chromatography. Analyzes were carried out by EAST, SDS-PAGE, isoelectric focusing, immunoblotting and amino-acid sequencing. RESULTS: An 18-kDa protein from the P. acerifolia pollen extract, which we named Pla a 1, was purified. This nonglycosylated protein had an isoelectric point value higher than 9.3 and was recognized by up to 92% of monosensitized Platanus allergic patients and 83% of polyzensitized patients. Sequencing of its N-terminal yielded an amino acid sequence which showed no homology to the known proteins in the databases. Other relevant allergens detected in monosensitized patients were proteins of 43 and 52 kDa, with immunoglobulin (Ig)E-binding prevalences of 83 and 42%, respectively. Profilin was an important allergen in polyzensitized patients. CONCLUSIONS: The most relevant allergens from the P. acerifolia pollen have been determined. A major allergen, specific of this pollen, and named Pla a 1, has been purified and characterized. PMID- 11906337 TI - Influence of food processing on the allergenicity of celery: DBPCFC with celery spice and cooked celery in patients with celery allergy. AB - BACKGROUND: Celery root is often consumed in a processed form as a cooked vegetable or as a spice. So far, however, there has been no information about the allergenicity of processed celery in celery-allergic patients. METHODS: In 12 patients with a history of allergic reactions to raw or raw and cooked celery, double-blind placebo-controlled food challenges (DBPCFCs) with raw celery (n = 10), cooked celery (110 degrees C/15 min; n = 11), and celery spice (n = 5) were performed. Nine patients underwent an open mucosal challenge with four samples of canned celery retorted at Co-values (cooking effect) of 7.45-76.07 (corresponding to the time periods in minutes at a thermal influence of 100 degrees C). IgE immunoblot analysis of celery extract was performed with sera of all challenged patients. The thermal stability of celery allergen was investigated by enzyme allergosorbent test (EAST) inhibition. Furthermore, intraperitoneal immunization of mice followed by a rat basophil leukemia (RBL) cell mediator release assay was used as a biological in vitro model to assess the allergenicity of processed celery. RESULTS: Six out of 11 patients showed a positive DBPCFC to cooked celery and five out of five patients to celery spice. Allergenicity of celery was preserved in four patients with a positive DBPCFC to cooked celery even if celery was treated at a Co-value of 76.07. Patients with positive DBPCFC to cooked celery reacted to known celery allergens (Api g 1, Api g 4, cross-reactive carbohydrate determinants CCD). EAST inhibition showed that heat resistance of celery allergens decreases in the following order: CCD > Api g 4 > Api g 1. Accordingly, five of six patients with a positive DBPCFC to cooked celery were sensitized to profilin and/or CCD. The murine model reflected the reactivity of patients sensitized to the major allergen Api g 1. CONCLUSIONS: 1) In a subset of patients with a positive DBPCFC to cooked celery, celery remains allergenic even after extended thermal treatment (76.07 min/100 degrees C). 2) Celery spice is allergenic for patients with an allergy to raw celery. 3) RBL cells sensitized with mouse IgE to raw celery may serve as a useful tool for screening the potential allergenicity of heat-processed products containing celery. PMID- 11906338 TI - Features and severity of occupational asthma upon diagnosis: an Italian multicentric case review. AB - BACKGROUND: The severity of occupational asthma (OA) at the time of diagnosis is not known. In this study we aimed to evaluate some features of the disease at the time of diagnosis, particularly looking at severity and treatment before diagnosis. METHODS: Medical records of subjects (n = 197) who had received a diagnosis of OA in six specialized centres of Northern and Central Italy in the period 1992-97 were reviewed. The severity of the disease at the time of diagnosis was determined on the basis of symptoms, peak expiratory flow (PEF, percentage predicted), forced expiratory volume in one second (FEV1, percentage predicted), and PEF variability, following the criteria of the National Institutes of Health and World Health Organizaton (NIH/WHO) guidelines on asthma. Medications used in the month before diagnosis were recorded. RESULTS: The most common etiological agents were isocyanates (41.6%), flours (19.8%), woods (9.7%) and natural rubber latex (7.6%). The level of asthma severity (AS) was mild intermittent in 23.9% patients, mild persistent in 28.9%, moderate in 41.6%, and severe in 5.6%. Asthma severity was positively associated with current or previous smoking (P < 0.05), and was not related to atopy and current exposure. A relationship with bronchial reactivity to methacholine was shown in subjects at work. Treatment before diagnosis was consistent with the NIH/WHO guidelines in only 13.2% patients, whereas 75.6% were undertreated and 11.2% were overtreated. CONCLUSIONS: In this study we found that the majority of patients had mild asthma at the time of diagnosis and that cigarette smoking was associated with a greater severity. Moreover, the majority of patients were undertreated before etiological diagnosis. PMID- 11906339 TI - No effect of oral treatment with an intestinal bacterial strain, Lactobacillus rhamnosus (ATCC 53103), on birch-pollen allergy: a placebo-controlled double blind study. AB - BACKGROUND: Oral probiotic bacteriotherapy with Lactobacillus rhamnosus has given promising results in small children with food allergy. We studied the effects of similar therapy in teenagers and young adults, who were allergic to birch pollen and apple food and had intermittent symptoms of atopic allergy and/or mild asthma. METHODS: We conducted a double-blind, placebo-controlled study, in which respiratory and eye symptoms and use of medications in two groups were compared. Open oral challenge tests with a slice of apple were performed trice: before, during and after the birch-pollen season. There were 18 patients in each group. They used Lactobacillus rhamnosus for 5.5 months; 2.5 months before the pollen season, 1 month during the season (May), and 2 months after. RESULTS: The results were negative. The treatment did not alleviate the symptoms of the patients or reduce their use of medication during the birch-pollen season or the subsequent 2 months. The treatment did not significantly affect the symptoms caused by apple in the oral challenge tests. CONCLUSIONS: We found no indication of a beneficial treatment effect in our patients. As the number of patients was relatively small, conclusions should be drawn with caution. PMID- 11906341 TI - IgE-mediated anaphylaxis to Thiomucase, a mucopolyssacharidase: allergens and cross-reactivity. AB - BACKGROUND: Thiomucase is a mucopolysaccharidase obtained from ovine tissues mainly used to facilitate the diffusion of local anaesthetics and in the treatment of cellulitis. A patient with an anaphylaxis in relation to the intramuscular administration of Thiomucase is reported. OBJECTIVE: To investigate Thiomucase allergens and their possible relationship with dander allergens and animal albumins. MATERIAL AND METHODS: Skin prick tests (SPT) and serum-specific IgE were performed with Thiomucase and danders. Thiomucase SDS-PAGE immunoblotting was performed in order to study allergens. RAST/CAP inhibition and SDS-PAGE immunoblotting inhibition were carried out to study the cross reactivity. RESULTS: Skin prick tests (SPT) were positive to Thiomucase, animal dander (cat, dog, sheep, other), bovine serum albumin (BSA), and echinococcus. Specific IgE was also positive to Thiomucase, animal dander (cat, dog, sheep, other), BSA and echinococcus. In the RAST-CAP inhibition assays BSA was nearly completely inhibited by Thiomucase, Thiomucase was partially inhibited by BSA and cat and Echinococcus granulosus was partially inhibited by sheep and Thiomucase. In the Thiomucase SDS-PAGE immunoblotting several proteins fixed IgE, ranging from 20 kDa to > 94 kDa, the strongest with 43 kDa. The IgE fixation to BSA, cat and sheep in the SDS-PAGE immunoblotting was completely inhibited by the preincubation of the serum with Thiomucase. CONCLUSIONS: An IgE-mediated anaphylaxis to Thiomucase is documented. Multiple allergens are recognized in Thiomucase by the patient serum, the main with 43 kDa. Partial cross-reactivity with BSA, cat dander and sheep dander is documented. PMID- 11906340 TI - The association between synthetic bedding and adverse respiratory outcomes among skin-prick test positive and skin-prick test negative children. AB - BACKGROUND: Synthetic bedding has been associated with increased child wheeze and also higher allergen levels in several studies. We aimed to examine whether the association between synthetic bedding and adverse respiratory outcomes was more evident among skin-prick test (SPT) positive children. METHODS: A cross-sectional survey involving a population sample of 758 (81% of eligible) school children aged 8-10 years from randomly selected schools in the Australian Capital Territory in 1999. Parental questionnaires for ISAAC respiratory symptoms and child bedding were obtained. SPT results of 10 common allergens were available on 722 of the subjects (77% of those eligible). Synthetic pillow or quilt use was termed synthetic upper bedding. RESULTS: Synthetic quilt use was associated with asthma (Adjusted Odds Ratio 1.67 (1.05, 2.65)), recent wheeze (AOR 1.63 (1.03, 2.59)) and allergic rhinoconjunctivitis (AOR 2.11 (1.33, 3.34)) among SPT positive children. However, these associations were not apparent for SPT-negative children. Similarly, increasing synthetic upper bedding use was associated with more than 12 episodes of wheeze among SPT-positive children (AOR 1.69 (1.08, 2.64), P=0.02, per category) but not SPT-negative children (AOR 0.77 (0.26, 2.21), P=0.6, per category). CONCLUSION: The apparent association between synthetic upper bedding and adverse respiratory outcomes was evident among SPT positive but not SPT-negative children. Prospective intervention studies that aim to examine the effect of upper bedding composition on child asthma among SPT positive children are required. PMID- 11906342 TI - Severe asthma and plasma serotonin. PMID- 11906343 TI - Cockroach debris in purchased flour. PMID- 11906344 TI - Food allergy to spinach and mushroom. PMID- 11906345 TI - Roxithromycin induced acute urticaria. PMID- 11906346 TI - Allergy to durian. PMID- 11906347 TI - Isolated allergy to rice. PMID- 11906348 TI - Anaphylaxis to celecoxib. PMID- 11906349 TI - The allergenicity of feather bedding material. PMID- 11906350 TI - Unusual soy oil allergy. PMID- 11906351 TI - Natural course of AEDS. PMID- 11906352 TI - Allergic contact eczema/dermatitis from cosmetics. PMID- 11906353 TI - T cell mediated allergy to abciximab. PMID- 11906354 TI - Pneumothorax following insect sting anaphylaxis. PMID- 11906355 TI - Long-lasting clinical efficacy of allergen specific immunotherapy. PMID- 11906356 TI - Tuberculin reactivity and allergy. PMID- 11906357 TI - Adverse respiratory effects and allergic susceptibility in relation to particulate air pollution: flirting with disaster. PMID- 11906358 TI - The role of histamine in allergic disease: re-appraisal of its inflammatory potential. PMID- 11906359 TI - The safety and efficacy of subcutaneous birch pollen immunotherapy - a one-year, randomised, double-blind, placebo-controlled study. AB - BACKGROUND: There is only very limited documentation of the efficacy and safety of high-dose subcutaneous birch pollen immunotherapy (IT) in double-blind, placebo-controlled (DBPC) studies. Birch pollen is a major cause of allergic morbidity in northern Europe and in eastern parts of North America. METHODS: Thirty-five patients with severe rhinoconjunctivitis (hay fever) to birch pollen were allocated to double-blinded clustered IT with a depot birch pollen extract (Betula verrucosa) or placebo injections. Seven patients in each group had concomitant self-reported seasonal asthma. Treatment was conducted as a clustered regimen and was performed in a specialist unit. Symptom scores from nose, eyes, and lungs, and use of oral and topical antihistamines, beta-2-agonists, and oral corticosteroids were recorded daily during the season of 2000. Sensitivity to allergen provocation in skin, conjunctiva, and nasal mucosa was measured before and after 10 months of treatment. Post-seasonal assessment of symptom severity was performed using a simple questionnaire. RESULTS: IT reduced the symptom score for both rhinoconjunctivitis and asthma (P-values < 0.05), total medication score (P < 0.02) and use of oral antihistamines (P < 0.01). IT reduced specific conjunctival sensitivity (P < 0.05), skin prick test, and especially cutaneous late-phase response diameters (P < 0.00001), and increased general well-being on post-seasonal evaluation (P < 0.01). IT was safe, with side-effects at the same level as placebo. CONCLUSIONS: High-dose, subcutaneous IT is efficacious and safe in patients with severe birch pollen rhinoconjunctivitis and asthma. PMID- 11906360 TI - Long-term efficacy of preseasonal grass pollen immunotherapy in children. AB - BACKGROUND: In a previous controlled study we demonstrated that preseasonal grass pollen immunotherapy for three years was effective in children. In the current study we examined the same group of patients to see if there is still a benefit six years after discontinuation of treatment. METHODS: Thirteen of 14 patients with previous specific immunotherapy (SIT) and 10 out of 14 patients of the control group were prospectively followed during the grass pollen season. Outcome measures were seasonal symptom scores for eyes, nose and chest, the use of symptomatic medication and visual analog scale. Objective measures included skin prick test reactivity to seasonal and perennial allergens and conjunctival provocation testing. RESULTS: During the 13 week observation time scores for overall hayfever symptoms (P < 0.004) and individual symptoms for eyes (P < 0.02), nose (P < 0.04) and chest (P < 0.01) as well as combined symptom and medication scores (P < 0.002) remained lower in the group with previous SIT. Only 23% of patients with previous pollen-asthma who had received SIT experienced pollen-associated lower respiratory tract symptoms compared to 70% in the control group (P < 0.05). There was no significant difference in the use of pharmacological treatment during the pollen season except for asthma medication. The average visual analog scale was lower in the post-SIT group (P < 0.05). Six years after cessation of SIT the immediate skin response to grass pollen remained decreased compared to the reaction of the controls (P < 0.01). There was also a tendency for higher allergen concentration to provoke a conjunctival response in the post-SIT group but without reaching statistical significance. Eight years after commencement of SIT, 61% of the initially pollen-monosensitized children had developed new sensitization to perennial allergens compared to 100% in the control group (P < 0.05). CONCLUSIONS: There is still a significant clinical benefit six years after discontinuation of preseasonal grass pollen immunotherapy in childhood. SIT in children with pollen-allergy reduces onset of new sensitization and therefore has the potential to modify the natural course of allergic disease. PMID- 11906361 TI - Allergenicity and immunochemical characterization of six varieties of Olea europaea. AB - BACKGROUND: The inhalation of Olea europaea pollen is one of the most important causes of allergic respiratory diseases in the Mediterranean basin. The objective of this study was to investigate the antigenic and allergenic composition of six different O. europaea varieties collected in southern Spain. METHODS: The varieties included in the study were: Acebuche (wild olive), Carrasqueno, Nevado, Hojiblanco, Manzanillo and Picual. Extracts of these six varieties were prepared. Twenty-nine olive individuals with an immunoglobulin(Ig)E-mediated allergy to olive pollen were skin tested with these extracts. The antigenic profile of these extracts was evaluated by SDS-PAGE; the allergenic profile was investigated by immunoblotting using the serum of these 29 individuals. The Ole e 1 content was established by ELISA inhibition using purified Ole e 1 and rabbit polyclonal antibodies and by scanning densitometry. RESULTS: The extracts that induced the smallest wheal size were Acebuche and Hojiblanco, being significantly different from the rest of the extracts. The antigenic and allergenic profiles of the extracts also varied. The Ole e 1 content ranged from 0.050 in Hojiblanco to 0.232 in Manzanillo, measured by ELISA inhibition and from 0.153 in Hojiblanco to 0.677 in Nevado, measured by scanning densitometry. CONCLUSIONS: The different varieties of O. europaea pollen studied demonstrated great differences in the in vivo and in vitro potency of the extracts. There were significant differences in the Ole e 1 content, while the protein content remained very similar in these extracts. This study confirms previous observations of a great variability in the antigenic and allergenic composition of O. europaea pollen extracts and establishes significant differences in Ole e 1 content. PMID- 11906362 TI - The spectrum of allergic (cross-)sensitivity in clinical patch testing with 'para amino' compounds. AB - BACKGROUND: Allergic contact sensitization to 'para amino' compounds is frequent and the spectrum of cross-reactivity between members of this chemical group is variable. METHODS: A retrospective analysis of clinical patch test data obtained with a special test series in the centres of the Information Network of Departments of Dermatology (IVDK) between 1995 and 1999. RESULTS: In the 638 patients tested with the above test panel positive reactions were observed most often to p-aminoazobenzene (16.2%), p-phenylenediamine (14.1%), p toluylenediamine (10.0%), followed by 4,4'-diaminodiphenylmethane (8.5%), Disperse Orange 3 (8.4%) and p-aminophenol (3.1%). Among the 544 patients tested with p-phenylenediamine and all seven additional 'para amino' compounds, concordance between reactions varied greatly. The stronger the positive test reaction to p-phenylenediamine, p-toluylenediamine or p-aminoazobenzene, the more frequently additional positive reactions to the other compounds were observed. CONCLUSIONS: A screening employing several 'para amino' compounds is necessary to describe the individual spectrum of allergic contact sensitization, as there is no reliable marker substance. Further research should aim at (i) establishing the mechanism of cross-reactivity to 'para amino' compounds and (ii) identifying exposures in the environment. PMID- 11906363 TI - Down-regulated IL-5 receptor expression on peripheral blood eosinophils from budesonide-treated children with asthma. AB - BACKGROUND: The expression and function of cytokine receptors on peripheral blood eosinophils (PBE) from healthy and asthmatic children are poorly characterized. METHODS: The PBE count and expression of IL-5 receptor (R) and GM-CSFR positive PBE was analyzed in nonsteroid-treated asthmatic children (n = 13), budesonide treated asthmatic children (n = 24) and healthy children (n = 16) by flow cytometry. Alterations in intracellular EG2-epitope expression were used to measure the in vitro responsiveness of PBE to recombinant IL-5 and GM-CSF. RESULTS: The PBE count was increased (P < 0.05) in both asthmatic groups, independent of treatment, as compared to healthy children. The IL-5R expression on PBE, as well as the in vitro responsiveness of PBE to recombinant IL-5, was reduced (P < 0.05), in budesonide-treated asthmatic children compared to nonsteroid-treated asthmatic children and healthy children. The proportion of GM CSFR positive PBE and in vitro responsiveness of PBE to recombinant GM-CSF were not different between the groups. In vitro treatment with budesonide did not down regulate the proportion of IL-5R positive PBE. CONCLUSIONS: Budesonide-treatment of asthmatic children induces a selectively reduced IL-5R expression on PBE, concomitant with a reduced in vitro responsiveness of PBE to IL-5. We suggest that this budesonide-related down-regulation of the IL-5R might be a mechanism by which steroid treatment inhibits the action of IL-5 on eosinophil accumulation and activation in vivo. PMID- 11906364 TI - Nasal lavage concentrations of free hemoglobin as a marker of microepistaxis during nasal provocation testing. AB - BACKGROUND: The constituents of nasal mucus may be contaminated by plasma if there is epistaxis. Gross epistaxis is apparent as a red lavage fluid, while microepistaxis may yield a clear fluid. If gross or microepistaxis are present, it will be difficult to decide whether plasma protein concentrations are elevated because of plasma exudation or bleeding. In order to discriminate between these two possibilities, we measured erythrocyte-derived free hemoglobin (fHb) in nasal lavage fluids. METHODS: Single-blinded subjects underwent standard hypertonic saline nasal provocation. Unilateral hypertonic nasal provocation was performed in normal, allergic rhinitis (AR) and nonallergic rhinitis (NAR) subjects (total of 1316 specimens). fHb was measured using the Sigma-Aldrich kit (St. Louis, MO). Grossly bloody specimens were analyzed separately from the remainder. Statistical analysis defined the means and 95th percentiles for fHb and albumin in the nonbloody normal group. RESULTS: fHb concentrations ranged from below the limits of detection (< 1 microg/ml) to > 164 microg/ml fHb was 79.3 microg/ml +/- 4.7 (mean +/- SEM) in four normal, 31 AR and 25 NAR grossly bloody specimens. The 95th percentile of fHb in the nonbloody normal samples (n = 68 subjects, n = 681 specimens) was 16.5 microg/ml. This value was defined as the threshold to detect potential microepistaxis, and corresponded to approximately 245 000 erythrocytes per ml of lavage fluid. Total protein (P < 0.05) and albumin (P < 0.001), but not markers of glandular secretion, were significantly increased in samples with fHb > 16.5 microg/ml compared to those < or = 16.5 microg/ml. Elevations of fHb without changes in albumin were more prevalent in nonallergic rhinitis. CONCLUSIONS: Significant bleeding into nasal lavage samples can contaminate the specimens and increase the concentrations of both fHb and plasma proteins. Increased albumin alone would indicate increased vascular permeability. The mechanism(s) leading to elevated fHb without increased plasma proteins require further investigation. PMID- 11906366 TI - The association of airway hyperresponsiveness and tuberculin responses. AB - BACKGROUND: The balance between the two subsets of T cell is pivotal for allergic sensitization. OBJECTIVE: We conducted a cross-sectional study of 486 children vaccinated with bacillus Calmette-Guerin (BCG), aged 10-13 years, to evaluate whether tuberculin responses may contribute to airway hyperresponsiveness (AHR). METHODS: Tuberculin skin test, allergic skin test, and methacholine challenge test were done. The methacholine concentration causing a 20% fall (PC20) in forced expiratory volume in 1 second (FEV1) was used as a threshold of AHR. Atopy was defined as a reaction showing a mean wheal size of > or = 3 mm to one or more allergens on skin prick test (SPT). Two tuberculin units of polysorbate stabilized purified protein derivatives (PPD) were injected intradermally into the volar surface of the forearm. Reactions were read at 48-72 h as the transverse diameter in millimeters of induration. RESULTS: Of the children in the study, 12.3% (60/486) had PPD induration; 7.8% (38/486) of children had PPD induration of greater than 10 mm. The PPD induration size was 10.5 +/- 1.03 mm (confidence interval (CI) 7.19-12.33) in atopic children and 11.2 +/- 0.76 mm (CI 7.89-13.1) in nonatopic children. The differences of PPD induration diameter between the two groups were not significant. There was no difference of log PC20 between PPD induration > or = 10 mm and < 10 mm (0.13 +/- 0.18 vs. 0.42 +/- 0.05). The difference of log PC20 between positive and negative tuberculin response was not significant. Children with atopy had lower log PC20 than those without atopy (0.16 +/- 0.07 vs. 0.51 +/- 0.05, P = 0.001). After adjusting for sex, age, height, weight, tuberculin response, atopy was associated with AHR in multivariate analyses (odds ratio = 1.895, CI 1.285-2.505, P = 0.002). CONCLUSION: These data suggested that a tuberculin response due to mycobacterial infection status have no effect on AHR in schoolchildren. PMID- 11906365 TI - Absence of relationship between tuberculin reactivity and asthmatic symptoms, level of FEV1 and bronchial hyperresponsiveness in BCG vaccinated young adults. AB - BACKGROUND: Some recent studies have suggested that bacillus Calmette-Guerin (BCG) vaccination or mycobacterial infection early in life is inversely related to asthma. We wondered if an increase in tuberculin reactivity was inversely related to commonly used indices of asthma in a population of young adults who were BCG vaccinated at age 14. METHODS: Men and women aged 20-44 years, randomly selected from the general population, were tuberculin tested with the epinephrine Pirquet method with Norwegian-produced synthetic medium tuberculin (n = 588). In addition they were interviewed using eight questions on asthma symptoms and medication. Lung function and bronchial responsiveness were also tested. RESULTS: Altogether 95% of those studied had been BCG vaccinated at age 14 (n = 558). In the 386 subjects with complete examinations, there was no relationship between a positive tuberculin reaction (> or = 4 mm) and asthma symptoms or use of asthma medication. Furthermore we did not observe any relationship between a positive tuberculin reaction and the level of forced expiratory volume (FEV1) or a positive bronchial responsiveness test, assessed as the percent of predicted of FEV1 and PD20 < 2 mg methacholine, respectively. In multiple logistic regression analyses neither respiratory symptoms, level of FEV1, nor bronchial hyperresponsiveness were related to tuberculin reactivity after adjustment for age, gender and smoking habits. CONCLUSIONS: In this young adult population who were BCG vaccinated at the age of 14 no significant relationship existed between tuberculin reactivity and asthmatic symptoms, level of FEV1 or bronchial hyperresponsiveness. Our data does not support the hypothesis that BCG immunization is beneficial in reducing asthmatic symptoms and disease in young adults. PMID- 11906367 TI - Comparison of five new antihistamines (H1-receptor antagonists) in patients with allergic rhinitis using nasal provocation studies and skin tests. AB - BACKGROUND: It was the aim of the authors to compare all of the latest second generation antihistamines and to see if there were significant differences in their efficacy. It is important for ENT specialists to know if these differences exist, as it is for general practitioners trying to choose between these drugs. METHODS: In 12 confirmed grass pollen allergic patients the authors performed nasal smears to asses eosinophilia, histamine/grass pollen skin tests, and grass pollen nasal provocation tests. All tests were performed before and after administration of one of five different antihistamines (cetirizine, loratadine, ebastine, fexofenadine, mizolastine) or placebo. The order of administration of antihistamines and placebo was randomised, and patients were not aware of which drug they were given. A decrease in nasal eosinophilia (nasal smear), or nasal or skin reactivity (provocation tests) was looked for. RESULTS: A significant decrease in nasal eosinophilia was observed for all antihistamines but not for placebo. For the grass pollen nasal provocation tests, the decrease was significant for nasal blockage and sneezing; for rhinorrhea there was an insignificant decrease that was true for all antihistamines. A significant reduction in histamine/grass pollen skin test reactivity was also observed for all antihistamines, during an 8 h observation period. A significant difference in efficacy between the different antihistamines could not be found with any of the tests performed. CONCLUSIONS: For the newer nonsedating H1-antagonists there appears to be no clinically relevant differences in activities--at least not in our study. Preference of the patient may be the most important factor in making a choice between these drugs. PMID- 11906368 TI - Platanus acerifolia pollinosis and food allergy. AB - BACKGROUND: In Mediterranean areas, oral allergy syndrome (OAS) occurs independently of an associated birch pollinosis; moreover, on occasions it presents with no other associated pollinosis. The aim of this study was to assess the possible association of OAS with Platanus acerifolia pollinosis. METHODS: We evaluated consecutive patients seen for pollinosis in an allergy department. Seven hundred and twenty patients were selected on the basis of seasonal or perennial rhinitis, or asthma, or both. Respiratory and food allergies were studied in all patients. Clinical history was recorded and examinations and skin prick tests were performed with a battery of available common inhalant allergens and plant-derived food allergens. Specific IgE levels to P. acerifolia pollen extract and food allergens tested were measured. Molecular masses of the IgE binding proteins and cross-reactivity among the P. acerifolia pollen and different food extracts were also determined. RESULTS: Of the 720 patients evaluated, 61 (8.48%) were sensitized to P. acerifolia pollen. Food allergy was observed in 32 (52.45%) of the 61 patients sensitized to P. acerifolia pollen. Food allergens most frequently implicated were hazelnuts, peach, apple, peanuts, maize, chickpea and lettuce. Enzyme allergosorbent (EAST)-inhibition showed high inhibition values when P. acerifolia pollen extract was used as free phase. On the contrary low inhibition was observed when plant-derived food allergens were used as free phase and P. acerifolia pollen extract as solid phase. CONCLUSIONS: Cross-reactivity was observed among P. acerifolia pollen and plant-derived foods. OAS in these patients may have been caused by primary respiratory sensitization. PMID- 11906369 TI - Allergic sensitization owing to 'second-hand' cat exposure in schools. AB - BACKGROUND: Environmental allergen loads play an important role in triggering symptoms in atopic individuals. While a number of previous studies have shown that cat allergens (Fel d 1) can be found in school dust samples, no study has provided evidence that public places contribute to increased atopic sensitization rates in children. METHODS: We employed data collected in a health survey of school children living in Germany in order to examine the association between the proportion of class- and schoolmates reporting cat contact and sensitization rates in children. RESULTS: Among 1893 children, 8.7% were sensitized to cats. Those sensitized were 5-7 times more likely to have received an asthma diagnosis or to have reported wheezing. Pupils without regular contact with cats were twice as likely to test positive for major cat allergen when the proportion of schoolmates with cat contact was high. No such relation was observed amongst children reporting regular cat contact. CONCLUSIONS: Our study suggests that allergens in school environments contribute to allergic sensitization and atopic diseases such as asthma. Thus, methods to reduce the allergen load in classrooms should be considered. PMID- 11906370 TI - Food allergy is a matter of geography after all: sesame as a major cause of severe IgE-mediated food allergic reactions among infants and young children in Israel. AB - OBJECTIVE: To determine the prevalence, importance, and the order of frequency of IgE-mediated food allergens among infants and young children in Israel. STUDY DESIGN AND PATIENTS: In a cross-sectional study, the prevalence of IgE-mediated food allergy was investigated in 9070 infants and young children (0-2 years) who were followed-up at 23 Family Health Centers (FHCs) in central Israel. Patients with suspected IgE-mediated food allergic reactions, were recruited for further evaluation (detailed questionnaire and skin-prick test (SPT)). RESULTS: We identified 150 out of 9070 (1.7%) patients with suspected IgE-mediated food allergy. Among them, 102/150 (67%) [59 males, 43 females; mean age 10.3 months] completed a detailed questionnaire and underwent SPT. Evaluation revealed 131 positive SPTs in 78/102 (76.5%) patients. Twenty-seven positive SPTs in 18 patients were considered clinically irrelevant based on previous consumption of the relevant foods without clinical symptoms. Thus, there were 104 relevant positive SPTs in 78 patients. The overall prevalence of clinically relevant IgE mediated food allergic reactions among these patients is estimated to be 1.2% (104/9070). The most common food allergens were egg, cow's milk, and sesame. Anaphylaxis was the presenting symptom in 14/78 (18%) including six sesame induced cases. A history of other atopic diseases was reported in 27 (35%) patients. In addition, 22 (28%) had a history of atopy in first-degree family members. CONCLUSIONS: We found sesame to be a major cause of IgE-mediated food allergy in Israel. In fact, it is second only to cow's milk as a cause of anaphylaxis. We recommend that testing for food allergens be tailored to each community based on local experience and should include sesame in appropriate populations. PMID- 11906371 TI - Chronic urticaria to atorvastatin. PMID- 11906372 TI - Efficacy of a questionnaire for allergy. PMID- 11906373 TI - Severe reaction on SPT. PMID- 11906374 TI - The ecNOS gene in allergic Czech children. PMID- 11906375 TI - Allergic anaphylaxis to Laminaria. PMID- 11906376 TI - Urticaria-angioedema by deflazacort. PMID- 11906378 TI - Primary colorectal signet-ring cell carcinoma. PMID- 11906377 TI - Oral desensitization to 5-ASA. PMID- 11906379 TI - Investigating rectal bleeding: red faced or reliable? PMID- 11906380 TI - Can axillary dissection be avoided in selected patients with breast cancer? PMID- 11906381 TI - Localization of the facial nerve in parotid surgery. PMID- 11906382 TI - Primary colorectal signet-ring cell carcinoma in Singapore. AB - BACKGROUND: Primary colorectal signet-ring cell carcinoma is a rare but distinctive tumour of the colon and rectum. The clinicopathological features are still controversial. The aim of this study is to review the clinicopathological features and management of this type of tumour in our hospital. METHODS: The clinicopathological features and survival data of all cases of primary colorectal signet-ring cell carcinoma were reviewed retrospectively. RESULTS: There were nine cases of primary colorectal signet-ring cell carcinoma in 3000 consecutive colorectal carcinoma patients seen from 1989 to 1999. There were seven male and two female patients with a mean age of 54.7 years. Three patients were younger than 40 years. The common presenting symptoms were rectal bleeding (33%) and small bowel obstruction (33%). Two (22%) patients required emergency surgery due to acute small bowel obstruction. The most common tumour location was the right colon (44%) followed by the rectum (33%). All nine patients presented at a very late stage of disease. A majority (77%) had Dukes' C disease while two (22%) had Dukes' D disease with distant dissemination. Peritoneal spread (33%) was the most frequent way of dissemination. There was no patient with liver metastases at the time of diagnosis and initial presentation. The mean survival time was 30 (range 5-108) months. The 5-year survival rate was 12%. CONCLUSIONS: Primary colorectal signet-ring cell carcinoma is frequently diagnosed late with a very poor prognosis. A high incidence of peritoneal seeding and low incidence of liver metastases appears to be a characteristic of signet-ring cell carcinoma of the colon and rectum. PMID- 11906383 TI - Young patients with colorectal cancer: how do they fare? AB - BACKGROUND: Younger patients with colorectal cancer (CRC) have long been thought to have a poorer prognosis than older patients. Recent overseas reports, however, have disputed this. The aim of the present study was to conduct a review of data on patients with colorectal cancer collected over a 29-year period at Princess Alexandra Hospital (PAH) to ascertain the outcome of a younger subset of patients at this hospital. METHODS: The PAH Colorectal Project records on 2495 patients with malignancies of the colon, rectum and anus who were treated and followed since 1971, were analysed to determine clinical presentation, treatment and outcome. A group of 61 patients with colo-rectal adenocarcinoma was identified who were aged less than 40 years at presentation. Their clinical data were then compared with the larger group of older patients. RESULTS: There were 30 male and 31 female patients in the younger group. A positive family history was the most consistent risk factor, present in 34% of patients. Despite this, only one patient out of 61 had been diagnosed as a result of a screening programme. The Australian Clinico-Pathological Stage (ACPS), histology and distribution of tumours corresponded to that of the older patients. The overall 5-year survival among younger patients was 53%. The 5-year survival rates in younger patients were better than that for older patients for ACPS A and B, reaching statistical significance for both of these stages. CONCLUSIONS: Our results indicate that younger patients with colorectal cancer have the potential to do just as well as older ones. With the influence of a family history of colorectal cancer being very apparent in this group, greater emphasis should be placed on an adequate screening programme for them. PMID- 11906384 TI - Does type of surgeon matter in rectal cancer surgery? Evidence, guideline consensus and surgeons' views. AB - One of the most obvious but controversial trends in contemporary surgical practice is that of subspecialization. There is a lack of definitive evidence that subspecialization improves cancer outcomes largely because previous research is compromised by confounding variables of referral practice, lack of standardized definitions of surgical skills and selection bias. Randomized controlled trials of generalized versus subspecialist surgical care are unlikely ever to be performed. The present study of surgeons' views about the role of subspecialization in the care of colorectal cancer patients demonstrates partisan reactions among surgeons themselves (89% response rate). Results of national audits will contribute to wider debate about surgical subspecialization in colorectal cancer. PMID- 11906385 TI - Training general practitioners in flexible sigmoidoscopy to screen for colorectal cancer. AB - BACKGROUND: A screening programme to detect polyps or early carcinoma would significantly reduce the mortality and morbidity of colorectal cancer (CRC). The aims of the present study were to evaluate: (i) the feasibility of training general practitioners in flexible sigmoidoscopy (FS) for CRC screening; (ii) the acceptability of screening by faecal occult blood testing (FOBT) and FS in asymptomatic standard risk Australians aged over 50 years; and (iii) the yield of such screening. METHODS: Subjects were recruited by general practitioner (GP) referral, newspaper advertisement or by a direct approach to retirement villages. Participants were mailed a FOBT kit and a prescreening questionnaire. Flexible sigmoidoscopy was performed by a GP supervised by an experienced endoscopist. Subjects then completed a second questionnaire. General practitioners were assessed after 50 unassisted procedures. RESULTS: A total of 264 individuals contacted the study coordinator; 169 were screened. Screening was accepted well by the participants. Fifteen per cent of subjects had polyps and 4% had a positive FOBT. Training in FS was adversely affected by the availability of resources. Three GPs completed 50 unassisted procedures over a 15-month period, but none were able to reliably assess the distal bowel. CONCLUSIONS: Although the three trainees and their supervisors did not consider that the GPs were adequately trained after 50 unassisted procedures, training was adversely affected by limited resources within the Victorian public hospital system. Screening by FOBT and FS was considered to be acceptable by the patients undergoing these procedures. Existing facilities are not adequate if GPs are to be trained in FS as part of a national CRC screening program. PMID- 11906386 TI - Investigating chronic, bright red, rectal bleeding. AB - BACKGROUND: Chronic, bright red, rectal bleeding is a common symptom in our community and the aetiology is frequently benign anal disease. The aim of the present study was to determine the efficacy of performing a flexible sigmoidoscopy on patients with chronic, bright red, rectal bleeding who are at low risk for colorectal neoplasia and who, on rigid sigmoidoscopy, are found to have an identifiable anal cause (e.g. haemorrhoids, fissure) for their bleeding. METHODS: A prospective study was conducted on patients presenting with chronic, bright red, rectal bleeding. Patients were considered at low risk for colorectal neoplasia if they fulfilled the following criteria: (i) less than 55 years of age; (ii) no past or family history of colorectal neoplasia or inflammatory bowel disease; (iii) no symptoms of altered bowel habit or abdominal pain; and (iv) a source of bleeding identified (e.g. haemorrhoids, fissure) on rigid sigmoidoscopy. All patients underwent a flexible sigmoidoscopy. RESULTS: Eighty two patients were entered into the trial, mean age 39 +/- 9 years (range: 22-55 years), and the ratio of men:women was 1.8:1. The anal cause of bleeding was haemorrhoids in 96%, and anal fissure in 4%. At flexible sigmoidoscopy, five patients were found to have adenomatous polyps. Rigid sigmoidoscopy missed diminutive neoplastic lesions in 6% of patients. CONCLUSIONS: Flexible sigmoidoscopy results in a low yield of colorectal neoplasia in patients presenting with chronic, bright red, rectal bleeding who are at low risk for colorectal neoplasia and who have an identifiable anal cause for their bleeding. PMID- 11906387 TI - Frequency and predictors of axillary lymph node metastases in invasive breast cancer. AB - BACKGROUND: The objectives of the present study were to evaluate the incidence and predictors of axillary lymph node metastases (ALNM) in patients with breast cancer, and to identify if axillary surgery could be safely omitted in selected patients. METHODS: Between January 1996 and May 2000, 492 patients underwent 501 axillary lymph node dissections (ALND). The incidence of ALNM was correlated with clinical and pathological characteristics by univariate and multivariate analyses. RESULTS: Axillary lymph node metastases were found in 41% (207/501) of cases. Univariate analysis showed that palpability of primary and axillary lymph node (ALN), pathological tumour size, grade, lymphovascular invasion (LVI) and multifocality or multicentricity were significant predictors of ALNM. By multivariate analysis, palpability of ALN, pathological tumour size, LVI and multifocality or multicentricity remained as independent predictors. Among the 431 cases without palpable ALN, no ALNM were found if the tumour was < or = 5 mm, non-multifocal or multicentric, and without LVI, or the tumour was a tubular or mucinous carcinoma < or = 15 mm (n = 21). The frequency of ALNM in the absence of the other risk factors was 11% (7/64) if the tumour size was > 5-10 mm, and 17% (19/113) if the tumour was > 10-20 mm. However, the incidence of ALNM was 72% for the 32 clinically node-negative cases with multifocal or multicentric tumour > or = 10 mm and LVI. Those patients with palpable ALN (n = 66) had a greater than 50% risk of ALNM. CONCLUSIONS: Routine ALND could be omitted in clinically node negative patients with either a < or = 5-mm, LVI-negative tumour, or a < or = 15 mm tubular or mucinous carcinoma. Axillary lymph node dissection is still useful for determining pathological nodal status in all other cases, and in most cases with palpable ALN, as a therapeutic manoeuvre. PMID- 11906388 TI - Treatment of the axilla in early breast cancer: past, present and future. AB - BACKGROUND: The optimal treatment of the axilla in early breast cancer is controversial. The present study reviews the pattern and predictors of regional recurrence (RR) and prognosis after RR in patients with early breast cancer treated by conservative surgery and radiotherapy (CS + RT). Implications of the results on current practice and future directions are explored. METHODS: Between 1979 and 1994, 1158 patients with stage I or II breast cancer were treated with CS + RT at Westmead Hospital. Two groups of patients were compared: 782 patients who underwent axillary dissection (axillary surgery group) and 229 patients who received radiotherapy (axillary RT group) as the only axillary treatment. At least 10 lymph nodes were dissected in 82% of the axillary surgery group. Of the women in the RT group, 90% received RT to the axilla and supraclavicular fossa (SCF) only and 10% also received RT to the internal mammary chain (IMC). RESULTS: With a median follow-up period of 79 months for the axillary surgery group and 111 months for the axillary RT group, 27 patients developed a RR (2.8% and 2.2%, respectively). Seven patients (0.9%) in the axillary surgery group and three patients (1.3%) in the axillary RT group developed a RR in the axilla (P, not significant). Of the patients with SCF recurrences, 14 (1.8%) were in the axillary surgery group and one (0.4%) in the axillary RT group (P, not significant). One patient in the axillary surgery group developed concurrent axillary and SCF recurrences, while a patient in the axillary RT group developed an IMC recurrence. Twenty (74%) of the 27 patients with a RR developed a concurrent or subsequent distant relapse (30% and 44%, respectively). In the pathologically node-positive patients, the axillary recurrence rate was higher in those who had less than five nodes removed (17%) than those who had 10 or more nodes removed (0%; P = 0.01). The SCF recurrence rate was higher in patients with four or more positive axillary nodes (9.5%) than in those with 0-3 positive nodes (1.5%; P = 0.003). CONCLUSION: Adequate treatment of the axilla by surgery or RT alone is associated with a low rate of RR. The incidence of distant relapse was substantial in patients who developed a RR, which gives emphasis to the importance of optimizing local-regional control. PMID- 11906389 TI - Initial experience of vocal cord evaluation using grey-scale, real-time, B-mode ultrasound. AB - BACKGROUND: To evaluate whether grey-scale, real-time, B-mode ultrasound (US) is a reliable alternative to nasopharyngoscopy for assessing vocal cord function post-thyroid and post-parathyroid surgery. METHODS: A prospective validation study was undertaken comparing grey-scale, real-time, B-mode vocal cord US with the standard of nasopharyngoscopy in 100 consecutive patients undergoing thyroid and parathyroid surgery between 1 February 1999 and 31 August 1999, with seven patients with known cord palsy. The sensitivity and specificity of grey-scale, real-time vocal cord US for the detection of vocal cord palsy when compared to the standard of nasopharyngoscopy was analysed. RESULTS: In the postsurgical group, there were six nerves (3.2% of the nerves at risk) transient and no permanent vocal cord palsies. US identified four of six transient palsies and reported two false negatives and three false positives. US identified four of seven cord palsies in the non-surgical group with known cord palsy. Analysis of the 107 combined patients showed US had sensitivity of 62% (8/13), specificity of 97% (91/94), a positive predictive value of 73% (8/11) and a negative predictive value of 95% (91/96) for detecting cord paralysis compared to the standard of nasopharyngoscopy. CONCLUSION: Despite the enthusiasm of earlier reports, our initial experience with grey-scale, real-time, B-mode US suggests it is not a reliable alternative to nasopharyngoscopy for assessing vocal cord function post thyroid and post-parathyroid surgery. Further recruitment of patients with known vocal cord palsy is required to confirm or refute these initial impressions. PMID- 11906390 TI - Acute epiglottitis in adults: the Royal Melbourne Hospital experience. AB - BACKGROUND: To examine the common presentations and management of acute epiglottitis in adults. METHOD: Retrospective clinical study of 17 consecutive adult patients who presented to the Royal Melbourne Hospital between January 1988 and December 2000 was undertaken. RESULTS: The mean patient age was 47 years (range 20-87 years) and the male-to-female ratio was 1.8 : 1.0. Peak incidence occurred in September during early spring. All patients presented with sore throat and dysphagia; however, respiratory distress was only noted in 65%. The most common signs were temperature and tachycardia. Four patients (23%) required endotracheal intubation, which was performed electively in three and as an emergency in one. Three of 14 blood cultures were positive, two yielded Haemophilus influenzae type b and one yielded Streptococcus mitis. One of the four throat cultures was positive for Haemophilus influenzae type b. Twelve patients underwent awake flexible laryngoscopy under topical anaesthetic as part of their initial assessment, and there were no complications associated with this procedure. There was no mortality. CONCLUSIONS: The diagnosis of acute epiglottitis in the adult population is difficult as respiratory distress may be absent. Patients who have a significant sore throat with no obvious aetiology should have direct visualization of their larynx by flexible laryngoscopy. Lateral X-ray of neck is of limited value. Once diagnosed, these patients should be hospitalized and monitored as airway obstruction may develop rapidly. PMID- 11906391 TI - Pyogenic liver abscess complicated by endogenous endophthalmitis. AB - BACKGROUND: Endogenous endophthalmitis is an inflammation of ocular tissues that can lead to deterioration of and loss of vision. Rarely, this can complicate the course of a patient with pyogenic liver abscess. METHODS: Over an 18-month period, 68 patients were treated for pyogenic liver abscesses. Three patients, all of whom were male and with diabetes, were diagnosed with a Klebsiella pneumoniae liver abscess complicated by endogenous endophthalmitis. Open surgical or percutaneous drainage of the liver abscess was undertaken and the symptomology and outcome of the endophthalmitis reviewed. RESULTS: There was no mortality in our series. Two patients presented with simultaneous abdominal and ocular symptoms and one patient had ocular symptoms 3 days after surgical drainage of the liver abscess. Despite aggressive treatment, all patients had permanent deterioration of visual function with one patient becoming blind and requiring evisceration of the infected eye. CONCLUSION: Ocular symptoms in patients treated for pyogenic abscesses must be dealt with urgently with an ophthalmologic consultation. Increased awareness of this complication and a high index of suspicion are paramount for salvage of visual function. PMID- 11906392 TI - Predicted impact on Victoria's ambulance services of a new major trauma system. AB - BACKGROUND: In 1999, a new major trauma system was proposed for the state of Victoria, Australia. The guidelines for the new system were aimed at delivering major trauma cases to definitive trauma care in the least time possible. The aim of the present study was to analyse the potential effect of this system on Victoria's ambulance services. METHODS: The present study modelled the workload of major trauma cases in Victoria's ambulance service for one year pre- and post introduction of the guidelines. Cases were analysed regarding whether their first hospital destination would change under the proposed guidelines, and, subsequently, whether they would require interhospital transport to a higher level trauma service. The impact on the ambulance services was modelled as annual changes in distances travelled due to predicted changes in hospital destinations. RESULTS: Analysis of the predicted changes indicated that, in general, Victoria's metropolitan and rural road ambulance crews would not be greatly affected. However, some metropolitan road crews may have to travel extra distances for up to 110 cases per year. The major impact was on air retrieval crews, where the annual number of interhospital transfers is predicted to increase from approximately 150 to 330. CONCLUSIONS: The present study demonstrated that most of the impact of a new trauma system on Victoria's ambulance services could be readily absorbed into the current workload. However, it also highlighted areas affected disproportionately within the ambulance services; in particular, air retrieval. Such studies are important to enable the effective implementation of new trauma systems. PMID- 11906393 TI - Surface landmarks of the facial nerve trunk: a prospective measurement study. AB - BACKGROUND: Facial nerve identification and preservation is the key to safe parotidectomy in most clinical situations. Traditional approaches to the main trunk have depended solely on internal landmarks but localization may further be enhanced if data are available on its position with reference to neighbouring external features. METHODS: Prospective measurements were taken on a series of parotidectomy patients intraoperatively. Using that spot on the main trunk 10 mm proximal to its bifurcation as the reference point, the depth from the skin and its cranio-caudal distance from the summit of the tragus were measured. RESULTS: Thirty-three patients were included, whose body mass index showed that they had average body build. The reference point nerve was 23.6 mm (SD = 5.1 mm) from the skin surface, and 18.8 mm (SD = 6.0 mm) caudal to the tragus. CONCLUSION: Measurements relating to the siting of the facial nerve trunk were obtained live at operation. This information should be helpful in the initial mobilization of the parotid gland from its posterior relations and in facilitating the classical internal landmarks in the 3-D localization of the main trunk, thereby allowing quicker and safer parotidectomy. PMID- 11906394 TI - Herbert Moran Memorial Lecture. Exploring the legacy of history. AB - Herbert Michael Moran was born on 29 April 1885 in Sydney and died on 20 November 1945. I never had the good fortune to meet him but in 1946, when I was a fourth year medical student in Brisbane, I purchased a small brown-coloured book called Beyond the Hill lies China. To this day I cannot recall what led me to that purchase but I have looked at it from time to time over the past years and found it increasingly evocative of my own student and early hospital days. I have drawn deeply from the previous Moran lectures, particularly Sir John Loewenthal's lecture in 1974, which revealed for me the greatest detail about the man himself. I am indebted to Colin Smith, the College Archivist, for information held in his files; and Michael Moran who is here today for a number of personal reminiscences as well as family memorabilia. Moran was intensely interested in history. He was the founder of the first society dedicated to the study of the history of medicine in Australia, and through this activity he met the physician Leslie Cowlishaw whom he introduced to the College with the aim of securing Cowlishaw's 'vast knowledge of medical history and (his) wonderful library'. By a nice coincidence the third Cowlishaw Symposium was held at the College just 3 days ago, and Moran's College legacy is thereby increasingly recognized. PMID- 11906395 TI - Distal pancreatectomy with preservation of the spleen and splenic vessels. AB - This article draws attention to the concept of distal pancreatectomy with splenic preservation including the splenic artery and vein. PMID- 11906396 TI - Towards an Australian Institute of Trauma Research: learning the lessons of history. AB - Although medical research in Australia in the 20th century has resulted in a reduction of approximately 95% in the death rate from the infectious and parasitic diseases, there has been no such beneficial outcome in road traffic accidents which, it is here suggested, were out of control from 1948 until 1970. In 1970 several concerned community groups (spearheaded by the Royal Australasian College of Surgeons) campaigned for the introduction of mandatory seat belt wearing. As a consequence of that initiative, and the virtual plethora of subsequent research activities, the road traffic accident rate has declined substantially in the last three decades. The death rate from drownings to toddlers (children under 5) was essentially unchanged until the 1970s when research into toddler pool deaths and the implementation of those research findings resulted in a downturn in toddler drownings in the last two decades. These two examples demonstrate the value of accident research and the implementation of research findings. By analogy with the contributions of the large medical research institutes, the creation of an Australian Institute of Trauma Research seems desirable and is here proposed. PMID- 11906397 TI - Appendicovesical fistula arising from appendiceal diverticulum suspected on barium enema. PMID- 11906398 TI - Massive spontaneous haemopneumothorax in a patient with haemophilia. PMID- 11906399 TI - Tracheo-oesophageal fistula following a fall. PMID- 11906400 TI - Acquired urachal pathology: presentation of five cases and a review of the literature. PMID- 11906401 TI - Glomus tumour: the other umbilicated lesion of the stomach. PMID- 11906402 TI - Art macabre: Is anatomy necessary? PMID- 11906403 TI - Art macabre: is anatomy necessary? PMID- 11906404 TI - Art macabre: is anatomy necessary? PMID- 11906405 TI - Art macabre: is anatomy necessary? PMID- 11906406 TI - Art macabre: is anatomy necessary? PMID- 11906407 TI - Art macabre: is anatomy necessary? PMID- 11906408 TI - Art macabre: is anatomy necessary? PMID- 11906409 TI - Art macabre: is anatomy necessary? PMID- 11906412 TI - The evolution of surgery for myasthenia gravis. PMID- 11906413 TI - Surgery for liver metastases: how many? PMID- 11906414 TI - The need of ICU admission after major head and neck surgery. PMID- 11906415 TI - Resecting large numbers of hepatic colorectal metastases. AB - BACKGROUND: Despite the widespread use of surgical resection as a treatment for hepatic colorectal metastases, the value of resecting more than three metastases remains controversial. It was the objective of this study to determine if resection of larger numbers of metastases affects patient survival. METHOD: The survival of 123 consecutive patients who underwent curative hepatic resection for colorectal metastases between 1989 and 1999 by a single surgeon was analysed retrospectively. Kaplan-Meier survival statistics and Cox regression were used to determine the factors that affected survival, and logistic regression was used to determine the factors that affected the risk of recurrence of hepatic disease. RESULTS: The median survival rate for the whole group of patients was 38 months, with 1, 3 and 5 year survival rates of 88%, 53% and 31% respectively. The survival rate of patients undergoing resection of four to seven metastases (n=22; 5 year survival=39%) was not significantly different to that of patients undergoing resection of one to three metastases (n=91; 5 year survival=30%), P=0.9. Age, sex, primary cancer site, hepatic disease distribution, resection margins and adjuvant hepatic arterial chemotherapy (HAC) did not affect survival. Local invasion of the hepatic metastases (relative risk (RR)=2.9; P=0.001) and hepatic disease recurrence (RR=2.1; P =0.007) were the only factors that independently affected survival. Local invasion of the hepatic metastasis was the only factor associated with an increased risk of hepatic recurrence (RR=2.8; P=0.03). Adjuvant HAC did not affect the risk of hepatic recurrence (RR=1.5, P=0.4). CONCLUSION: Although there are no randomized trials that quantify any survival benefit from resection of liver metastases, the comparison of our results with well documented historical evidence indicates that surgical resection of up to seven colorectal liver metastases can result in a significant survival benefit. PMID- 11906416 TI - Retrospective study on the need of intensive care unit admission after major head and neck surgery. AB - BACKGROUND: The present article aims to study the pattern and need of Intensive Care Unit admission after major head and neck operations. METHODS: A retrospective study was undertaken of the hospital records of patients who underwent major head and neck operations during the period from February 1997 to February 2000 at the Division of Head and Neck Surgery, Department of Surgery, the Chinese University of Hong Kong. RESULTS: A total of 268 consecutive elective major operations were carried out over the 3 year period. The patients' age ranged from 14 to 82 years with a mean of 55 years. The male to female ratio was 4:1. Forty-seven patients underwent an operation with a combination of major resection, neck dissection, flap reconstruction and tracheostomy ('flaps group'). Two hundred and twenty-one patients had major head and neck operations without the need of flap reconstruction ('non-flaps group'). Three (6.3%) out of 47 patients (flaps group) were admitted to intensive care unit (ICU) immediately after the operation. Only one patient (2.2%) out of the remaining 44 patients was admitted for emergency treatment 3 weeks post operation. All four patients recovered uneventfully. In the non-flaps group of 221 patients, there were 12 (5.4%) planned admissions and 2 (0.96%) unplanned admissions to ICU. In the group of planned admissions, one out of the 12 patients died. The other two patients who were not planned for ICU admission died of basal meningitis that was disease related rather than related to the intensity of postoperative care. The overall admission rate to ICU was 18 (6.7%) out of 268 patients. The overall mortality was 1.1% (one planned, two unplanned). CONCLUSION: The present study showed that it is safe and cost-effective to discharge the majority of patients after major head and neck operations back to a specialist ward for nursing care. PMID- 11906417 TI - Optimal position for a cervical collar incision: a prospective study. AB - BACKGROUND: The descriptions of the optimal method for placing a cervical collar incision for thyroidectomy or parathyroidectomy are varied. It has been our impression that a collar incision marked in the neutral upright neck, migrates superiorly relative to the sternal notch when the patient is placed in the supine position. The aim of this study was to investigate the validity of this impression and to assess whether this is influenced by patient factors and/or pathology. METHODS: Fifty patients undergoing either thyroid or parathyroid surgery had a planned incision marked 1 finger-breadth (17 mm) above the sternal notch when sitting in the upright position. When placed in the supine position, with neck extended, the distance from the sternal notch to the marked incision was remeasured. Patient variables such as body mass index, height, weight and neck circumference were documented prospectively and data were recorded on operative details and tumour pathology. RESULTS: The collar incision marked in the neutral upright neck, migrated on average 21 mm superiorly once the patient was placed supine with the neck extended (P=0. 0001 ).The extent of migration was independent of all patient factors, type of operation and thyroid or parathyroid pathology. CONCLUSIONS: Migration of a proposed cervical collar incision does occur. An inappropriately placed incision may lead to excessive scarring if it is too low, or unusual prominence if it is too high. We believe a good position for marking such an incision is 1 fingerbreadth above the sternal notch with the patient in a neutral, upright position. PMID- 11906418 TI - Effectiveness of intra-operative wound infiltration with long-acting local anaesthetic. AB - BACKGROUND: Postoperative pain relief is of importance to both patients and surgeons. One of the simpler techniques is infiltration of the surgical incision with long acting local anaesthetic. The literature is confusing, with numerous reports attesting to the value of this approach and a similar number disputing the demonstrable benefits. METHODS: A prospective, 'blinded', randomized trial was undertaken involving 18 females undergoing uncomplicated bilateral sapheno femoral ligation. They received intraoperative bupivacaine infiltration into one or the other of their groin incisions. Postoperatively these patients were asked to assess a number of variables relating to their postoperative pain, each patient thereby acting as their own control. RESULTS: There was no significant difference in the postoperative pain experience between groins infiltrated with bupivacaine and those not infiltrated. Two patients developed wound infections and both of these occurred on the sides which had been infiltrated with bupivacaine, however this was not statistically significant. There was no difference found in pain experience with respect to pre- versus post-incision infiltration. CONCLUSION: Intraoperative wound infiltration with bupivacaine is widely employed as a method of minimizing postoperative pain. This study was unable to demonstrate a benefit of employing the technique in terms of a reduction in the patient's perception of pain. By having each patient act as their own control, we have avoided one of the major deficiencies of previous studies, namely, interpatient variability in assessment and perception of pain. PMID- 11906419 TI - Are there commonalities among audit tools used to measure carotid endarterectomy outcomes in New South Wales? AB - BACKGROUND: To prepare for a state-wide audit of carotid endarterectomy (CEA), we sought to ascertain pre-existing practices and identify common process and outcome measures among audits tools used by NSW vascular surgeons. METHOD: Telephone survey comprising five questions about CEA audit practices. RESULTS: Of 37 known vascular surgeons in NSW, all performed CEA and agreed to participate in our telephone survey (response rate 100%). All but one reported collecting CEA data (97%). Two thirds of these collected data prospectively. From those 19 surgeons (51%) who stated that they used standard tools for audit, we received five CEA-specific tools. Process measures common to all five were: side of CEA, date of operation, sex, past history of hypertension, history of diabetes, use of shunt, use of patch. There were only three outcome measures common to all five CEA-specific tools: postoperative transient ischaemic attack (TIA), postoperative stroke, and postoperative death. Data about outcomes beyond discharge were inconsistently collected. CONCLUSIONS: While most vascular surgeons in NSW report collecting information about CEA outcomes, not all do so prospectively. Only three outcomes (postoperative TIA, postoperative stroke and death) were common to all tools, inviting the development and use of a comprehensive and standard tool. PMID- 11906420 TI - Management of acute superior mesenteric artery occlusion. AB - BACKGROUND: This review examines the surgical management of acute superior mesenteric artery (SMA) occlusion and the impact of interventional radiology techniques. METHODS: Eight consecutive patients with SMA occlusion were treated at the Lismore Base Hospital, Lismore, NSW, Australia, from 1996 through to 2001 and of these, one patient was managed successfully with catheter-directed lytic therapy. The study group included five male and three female patients with a mean age of 71.3 (range 57-88) years. The records of these patients were reviewed to determine demographic characteristics, clinical features, predisposing factors and the duration of symptoms before intervention, management details and final outcome. RESULTS: Embolic phenomena due to atrial fibrillation were the most frequently identifiable cause of acute SMA occlusion, present in six of eight patients. Seven patients were managed with open surgery in the first instance and of these, four died. Three patients remain alive and well at a mean 2.8 years follow-up. Patient number eight developed acute SMA occlusion from embolism secondary to atrial fibrillation and was managed initially with SMA urokinase thrombolysis. This patient's pain was relieved 1 h after initiation of the procedure. Delayed films after 18 h from initiation of thrombolysis demonstrated re-opening of all the ileo-colic branches and at 6 weeks' follow-up the patient remains well with normal bowel function. CONCLUSIONS: There is a role for selective SMA cannulation and urokinase thrombolysis in the management of patients with acute SMA thrombosis. PMID- 11906421 TI - Virtual reality simulators: current status in acquisition and assessment of surgical skills. AB - Medical technology is currently evolving so rapidly that its impact cannot be analysed. Robotics and telesurgery loom on the horizon, and the technology used to drive these advances has serendipitous side-effects for the education and training arena. The graphical and haptic interfaces used to provide remote feedback to the operator--by passing control to a computer--may be used to generate simulations of the operative environment that are useful for training candidates in surgical procedures. One additional advantage is that the metrics calculated inherently in the controlling software in order to run the simulation may be used to provide performance feedback to individual trainees and mentors. New interfaces will be required to undergo evaluation of the simulation fidelity before being deemed acceptable. The potential benefits fall into one of two general categories: those benefits related to skill acquisition, and those related to skill assessment. The educational value of the simulation will require assessment, and comparison to currently available methods of training in any given procedure. It is also necessary to determine--by repeated trials--whether a given simulation actually measures the performance parameters it purports to measure. This trains the spotlight on what constitutes good surgical skill, and how it is to be objectively measured. Early results suggest that virtual reality simulators have an important role to play in this aspect of surgical training. PMID- 11906422 TI - The internet: from basics to telesurgery. AB - The exponential growth in information technology is resulting in a rapid increase in the ability to develop useful applications on the Internet. The purpose of the present article is to provide a brief review of the Internet with a consideration of its relevance to surgeons. This review is intended to indicate a range of relevant issues, rather than to discuss any specific topic in depth. It is becoming difficult for surgeons to reach their full potential unless they exploit Internet-based activities. This is because the ability to rapidly capture information of quality is an essential ingredient in a reflective approach to surgical problems. More futuristic is the prospect of using computer-based technology to operate upon patients from a distance (telesurgery). PMID- 11906423 TI - Paradigm shift in surgical approaches to thymectomy. AB - BACKGROUND: Thymectomy is an established therapy in the management of myasthenia gravis (MG) used in conjunction with medical treatment. The optimal surgical approach to thymectomy, however, has remained controversial. METHOD: The present review discusses the author's experiences of and the literature regarding the management of MG using the video-assisted thoracic surgery (VATS) approach. RESULTS: This approach was shown to be technically safe in experienced hands and associated with less postoperative pain, better preservation of lung function in the early postoperative period and better cosmetic results than alternative techniques. The intermediate term results of VATS are comparable to those of more radical approaches. It is hoped that this patient-friendly approach will lead to greater support by patients and their neurologists, for earlier surgery. CONCLUSION: VATS is an attractive, alternative approach to thymectomy. PMID- 11906424 TI - Recurrent torsion after previous testicular fixation. AB - BACKGROUND: Occasionally patients present with acute testicular pain having undergone a previous testicular fixation for torsion. The aim of this article is to review the literature and determine whether recurrent torsion of the testis is possible and whether the technique used at initial fixation is relevant. METHODS: A literature search was performed using MEDLINE, Pre-MEDLINE, EMBASE and CIANHL databases using the terms 'spermatic cord torsion' and 'testicular torsion' in combination with 'treatment failure', 're-operation' or 'recurrence'. RESULTS: Twenty cases of recurrent testicular torsion after fixation are described. Fifteen of the 17 that specified the type of suture used at initial fixation had used absorbable suture. Animal models suggest that to produce dense adhesions at testicular fixation, the tunica vaginalis must be everted behind the testis with or without fixation, before the testis is returned to the scrotum. Without this eversion, the adhesions around the testis are fine and only located at the site of fixation suture. The use of non-absorbable suture led to abscess formation in 30% of testes in animal models. CONCLUSIONS: The higher incidence of recurrent torsion after fixation using absorbable rather than non-absorbable sutures in the literature may be caused by a greater number of fixations being carried out using absorbable suture. Use of non-absorbable suture is limited by the high rate of abscess formation. The most important factor for adhesion formation would appear to be the eversion of the tunica vaginalis and it is recommended that this is carried out at all testicular fixations. PMID- 11906425 TI - Julian Smith: scientific surgeon, photographer, inventor. AB - Julian Augustus Romaine Smith was one of the surgeons who, in the company of Thomas Dunhill, Hugh Devine, Douglas Shields and David Murray Morton, established Saint Vincent's Hospital, Melbourne, as arguably the premier surgical hospital in Victoria, if not Australia. Smith abandoned a most promising scientific career to study medicine, completing his medical course in Melbourne where he graduated top of his year in 1898/1899. He gained his MD in 1901 and set up practice in Morwell, a Victorian country town, where he initiated his surgical career. In 1905 Smith returned to Melbourne and worked as assistant to Mr F. D. Bird, a prominent surgeon. He travelled overseas in 1906, visiting leading medical centres in England where Almroth Wright's work on vaccination made a deep impression on him. On his return to Australia in 1908 he joined the surgical staff at Saint Vincent's Hospital as surgeon to outpatients, finally retiring as surgeon to inpatients in 1928. Smith was made a Foundation Fellow of the Australasian College of Surgeons in 1927. Smith was regarded by his peers as a brilliant innovative surgeon with a special interest in urology. He became an expert cystoscopist. After he retired, he continued his long-standing interest in portrait photography for which he was considered a master. During World War II he designed and built an elegant roller pump for use in direct blood transfusion. He died in 1947, survived by his wife, a daughter and three sons, one of whom became President of the Royal Australasian College of Surgeons. PMID- 11906426 TI - Computer-based logbook for surgical registrars. AB - BACKGROUND: The documentation and monitoring of operative experience is an important component of advanced surgical training. The Royal Australasian College of Surgeons (RACS) monitors the adequacy of training by use of the surgical logbook. The logbook has been a paper-based record that does not permit longitudinal evaluation of the progress of an individual trainee or comparison of different surgical units. METHODS: An electronic logbook has been developed in FileMaker Pro version 5.03 (FileMaker, Santa Clara, California, USA). RESULTS: The electronic logbook has been employed for 1 year and has been used on both Windows and Macintosh platforms without difficulty. Appropriate summaries of the training experience were provided for the RACS at the conclusion of each rotation. CONCLUSIONS: The use of a relational database for logbook purposes provides trainees with a convenient and versatile record of their experience while meeting RACS requirements for documentation of surgical experience. PMID- 11906427 TI - Primary malignant melanoma of the oesophagus presenting with massive melena and hypovolemic shock. PMID- 11906428 TI - Primary squamous cell carcinoma of the breast. PMID- 11906429 TI - Benign cystic teratoma of the omentum. PMID- 11906430 TI - Left paraduodenal hernia: case report and review of the literature. PMID- 11906431 TI - Tension haemothorax: an uncommon life-threatening complication. PMID- 11906432 TI - SCARS data inadequate to support conclusion. PMID- 11906433 TI - Eosinophilic colitis. PMID- 11906436 TI - Breast cancer prevention: an opportunity for better understanding of time challenged risk factors. PMID- 11906437 TI - Mammary gland architecture as a determining factor in the susceptibility of the human breast to cancer. AB - The developmental pattern of the breast can be assessed by determining the composition of the breast in specific lobular structures, which are designated as lobules type 1 (Lob 1), lobules type 2 (Lob 2), and lobules type 3 (Lob 3), with Lob 1 being the less developed and Lob 3 being the most differentiated or with the highest number of ductules per lobular unit. In the present work, the patient population consisted of three groups of women who underwent surgical procedures: The first group included women who underwent reduction mammoplasty (RM) for cosmetic reasons. The second group included women who underwent prophylactic subcutaneous mastectomy after genetic counseling for either carrying the BRCA-1 gene or belonging to a pedigree with familial breast cancer (FAM), and the third group included women who underwent modified radical mastectomy (MRM) for the diagnosis of invasive carcinoma. The RM group consisted of 33 women, of whom 9 were nulliparous and 24 were parous. The FAM group consisted of 17 women, of whom 8 were nulliparous and 9 were parous. The MRM group consisted of 43 women, of whom 7 were nulliparous and 36 were parous. The analysis of the lobular composition of all of the samples from the RM group, which is considered the control group, revealed that Lob 1 represented 22%, Lob 2 represented 37%, and Lob 3 represented 38%, whereas the tissue examined from the FAM and MRM groups contained a preponderance of Lob 1 at 48% and 74%, respectively, over Lob 3, which was 10% and 3%, respectively. When the results of the analysis of breast tissue were separated according to the pregnancy history of the donor, it was found that in the control group or RM, there was a significant difference in lobular composition. Nulliparous women of the RM group showed a preponderance of Lob 1 (46%) over parous women, which contained only 17%, whereas the percentage of Lob 3 in the nulliparous group was significantly lower (7%) than the parous group (48%). In the breast tissues obtained from FAM and MRM, no significant differences in lobular composition were observed, as all of the samples contained a higher concentration of Lob 1, independent of the pregnancy history. The breast tissue of FAM and MRM of parous women had a developmental pattern that was similar to that of nulliparous women of the same group and that was less developed than the breast of parous women of the control group. An important difference between the Lob 1 of the FAM group versus the control (RM) and the MRM group was that most of these lobules had thin ductules with an increase in hyalinization of the intralobular stroma manifested in the whole-mount preparation as an alteration in the branching pattern. The data suggest that the breast tissue of women with invasive cancer, as well as those from a background of familial breast cancer, have an architectural pattern different from the control or normal tissues and that the BRCA-1 or related genes may have a functional role in the branching pattern of the breast during lobular development, mainly in the epithelial stroma interaction. PMID- 11906438 TI - In situ duct carcinoma of the breast: clinical and histopathologic factors and association with recurrent carcinoma. AB - There has been a recent increase in the diagnosis of in situ duct carcinoma of the breast (DCIS) as a result of mammographic screening. DCIS is heterogeneous in appearance and likely in prognosis. There is no generally accepted model to predict progression to invasive carcinoma. We investigated the prognostic effect of clinical presentation and pathologic factors for women diagnosed with primary DCIS. A cohort of 124 patients was accrued between 1979 and 1994 and was followed to 1997; 78 had DCIS detected mammographically, and 88 underwent lumpectomy alone. In this article, we provide details about characteristics affecting the choice of primary therapeutic modality, and we examine the effects of factors on progression for the two patient subgroups. Presentation with bloody nipple discharge was associated with a significant increase in DCIS recurrence (p=0.07). The pattern of duct distribution was important: DCIS in which the involved ducts were more widely separated had a significantly greater recurrence of DCIS than when the involved ducts were more concentrated (p=0.08 for mammographically detected DCIS, p=0.07 for patients who underwent lumpectomy alone). For mammographically detected DCIS, younger patients had more DCIS recurrence (p=0.07). We found considerable heterogeneity in nuclear grade; 50% of patients exhibited more than one grade. Nuclear grade, necrosis, and architecture were not significantly associated with either recurrence of DCIS or development of invasive carcinoma. Longer follow-up will allow further evaluation of the prognostic relevance of the factors assessed. PMID- 11906439 TI - Importance of missed axillary micrometastases in breast cancer patients. AB - Axillary lymph node metastases dramatically worsen the prognosis of patients with breast cancer. Despite this prognostic significance, routine histologic examination of axillary lymph nodes examines less than 1% of the submitted material. It is therefore obvious that micrometastatic disease is missed with this rather cursory examination, and the question arises as to the significance of this missed disease. Most lines of evidence suggest that missed axillary micrometastases exist and contribute to patient mortality. Most large studies of breast cancer micrometastases have suggested that undetected axillary micrometastases can be identified with more detailed examinations of the regional lymph nodes and that this group of patients has a poorer prognosis than those with no metastases identified. In addition, small-volume nodal disease, too small to be detected by traditional hematoxylin and eosin staining, has been shown to be capable of producing tumors in animal models. Finally, micrometastases have been shown to be of significance in other diseases. This article reviews the lines of evidence and the ongoing studies that are attempting to clarify the significance of micrometastatic disease in patients with breast cancer. PMID- 11906440 TI - The current treatment of ductal carcinoma in situ. AB - A consensus conference was held in April, 1999, to help sift through the maze of controversy surrounding the treatment of mammographically detected ductal carcinoma in situ (DCIS). Members of the panel included approximately 30 DCIS experts, who addressed issues relating diagnosis, treatment, treatment of breast (and axilla), adjuvant therapy, among others. The panel agreed that the goal of treatment for DCIS is breast conservation and attempted to divide the population of patients with DCIS into subsets who are appropriately treated by mastectomy, radiation therapy, or by excision alone. Major criteria for breast conservation include small size of area of DCIS, clear surgical margins, and favorable biology. Neither axillary dissection nor sentinel node biopsy is appropriate for DCIS treated by breast conservation. The role of tamoxifen is currently under study, and although approved by the FDA for "risk reduction," its use in patients with DCIS is uncertain. PMID- 11906441 TI - Chemoprevention for high-risk women: tamoxifen and beyond. AB - The demonstration by the National Surgical Adjuvant Breast Project (NSABP) that 5 years of tamoxifen therapy is associated with an approximate 50% reduction in breast cancer incidence in high-risk women was a milestone in breast cancer prevention. Because tamoxifen is associated with increased risk of side-effects such as hot flashes, menstrual abnormalities, uterine cancer, and thromboembolic phenomena, its use will not be advisable or acceptable for all high-risk women. Women over 50 years of age appear to be at highest risk for serious adverse events, such as uterine cancer and thromboembolic phenomena. Individuals in whom tamoxifen-associated breast cancer risk reduction appears to outweigh risk of serious side-effects include women with prior in situ or estrogen receptor (ER) positive invasive cancer, atypical hyperplasia, and/or women ages 35-49 with a calculated Gail 5-year risk of > or =1.7%, hysterectomized women aged 50 and older with a 5-year Gail risk of > or =2.5%, and nonhysterectomized women aged 50 and older with a 5-year Gail risk of >5.0%. It is not yet clear whether tamoxifen can reduce breast cancer incidence in women with BRCA1 and BRCA2 mutations, although preliminary evidence favors benefit for at least those with a BRCA2 mutation. Raloxifene is a selective ER modulator with less uterine estrogen agonist activity than tamoxifen, and it is hoped that it will result in fewer uterine cancers but will be equally efficacious in reducing the risk of breast cancer. The NSABP is currently conducting a randomized study of tamoxifen versus raloxifene in high-risk postmenopausal women. Approximately one third of invasive cancers are ER negative. Tamoxifen does not reduce the incidence of ER-negative cancers, nor does it appear to be effective in preventing the appearance of one third of ER-positive cancers. Priorities in prevention research are to develop (a) biomarkers to refine short-term risk assessments based on epidemiologic models, (b) biomarkers predictive of response to specific classes of preventive agents, (c) drugs with fewer side-effects and/or effective in ER-negative or ER positive tamoxifen-resistant precancerous disease, and (d) efficient clinical trial models to assess new agent efficacy. Breast intraepithelial neoplasia (IEN) may be sampled by minimally invasive techniques and is an attractive short-term risk biomarker. Molecular abnormalities observed in IEN may be used to select potential agents for testing/therapy, and modulation of these abnormalities may be used in phase I trials to select appropriate doses and in phase II trials to assess response. Breast density volume and certain serum markers such as insulin like growth factor-1 are also being studied as potential risk and response biomarkers. Reversal or prevention of advanced IEN as well as modulation of other risk biomarkers in randomized phase II and phase III trials is being evaluated as a means of more efficiently evaluating prevention drugs in the future. A number of agents are being developed that target molecular abnormalities in IEN, have fewer or different side effects than tamoxifen, and may be effective in ER negative or tamoxifen-resistant disease. PMID- 11906442 TI - Prophylactic surgery to reduce breast cancer risk: a brief literature review. AB - Prophylactic mastectomy reduces the likelihood of developing breast cancer among women at heightened risk for breast cancer, but at significant personal cost. Women at increased breast cancer risk on the basis of hormonal history, family history and/or genetic mutation carrier status may consider bilateral prophylactic mastectomy with or without reconstruction to reduce their cancer risk and/or decrease their chances of cancer mortality. Women having received mastectomy as treatment for breast cancer may request contralateral mastectomy to decrease the chances of developing a second breast primary. The potential oncologic value of these procedures must be weighed carefully on a case-by-case basis against the operation's physical and psychological morbidity. The purpose of this literature review is to provide a practice-oriented summary of recent clinical studies attempting to address the relative risks and benefits of preventive surgery for breast cancer. Data are included regarding the psychological factors surrounding patient selection and quality of life outcomes, which become the cornerstone of patient satisfaction and acceptance. Taken together, these data support the Society of Surgical Oncology position statement regarding the proper application of prophylactic surgery for breast cancer. PMID- 11906443 TI - The location of contralateral breast cancers after radiation therapy. AB - Radiation therapy following conservative surgery results in scattered radiation to the contralateral breast, with higher doses to the medial breast and lower doses laterally. The purpose of the current study is to determine whether the location of contralateral breast cancers developing following breast conserving surgery and radiation is indicative of radiation-induced malignancies. The charts of 1,755 patients treated with conservative surgery and radiation therapy between 1970 and 1998 were reviewed. Fifty-nine patients who developed a contralateral malignancy following conservative surgery and radiation therapy and who had complete information and documentation of the location of the second lesion served as the primary focus of the current study. The location of the contralateral malignancy was compared with the location of the primary tumors of the overall patient population. The location of breast cancers developing in the contralateral breast following breast conserving therapy and radiation was not consistent with radiation-induced malignancies. Specifically, there was not a preponderance of medially located tumors in patients developing contralateral breast cancers following radiation. There was a slight excess of central lesions that cannot be explained by higher doses of radiation. The location of breast cancers in the contralateral breast following conservative surgery and radiation is not indicative of radiation-induced lesions. These data should be reassuring to women considering breast conserving surgery and radiation. PMID- 11906444 TI - Her-2/neu gene amplification in low to moderately expressing breast cancers: possible role of chromosome 17/Her-2/neu polysomy. AB - Overexpression of the Her-2/neu (HER2) oncogene is known to confer important prognostic and predictive value to patients with breast cancer. Controversy exists as to the best method for its determination caused primarily by the variable sensitivities of the different antibodies and interobserver differences, particularly in the group of breast cancers with borderline levels of expression of the protein product. This study was therefore designed to determine the status of the HER2 gene amplification in a group of breast carcinomas with low levels of overexpression. After an initial validation of our procedures, a series of 52 consecutive cases of formalin-fixed, paraffin-embedded breast cancers with low levels of overexpression and a series of 22 cases with no expression by immunohistochemistry were analyzed by fluorescence in situ hybridization (FISH), and the results correlated statistically. Amplification of the HER2 gene was observed in 16% of equivocal to weakly positive cases. Those that were amplified showed low levels of amplification with ratios less than 4.5 and a characteristic scattered pattern of distribution of HER2 signals in the FISH assay. In addition, heterogeneity was noted in two cases in the amplification of the HER2 gene within the same tumor samples with pockets of amplified tumor cells amidst nonamplified tumor cells. In cases without amplification, a statistically significant number showed chromosome 17 polysomy. In conclusion, equivocal to low levels of HER2 overexpression in breast cancers are associated, in the majority of cases, with chromosome 17 polysomy and a corresponding increase in the HER2 gene numbers. True gene amplification is present in only a minority of cases. FISH analysis should be used for confirmation of gene amplification. Prior screening and selection of appropriate immunohistochemistry-positive areas for FISH analysis may prove beneficial. PMID- 11906445 TI - Spiritual expression and immune status in women with metastatic breast cancer: an exploratory study. AB - This exploratory study examined relationships between spirituality and immune function in 112 women with metastatic breast cancer. Spirituality was assessed by patient reports of frequency of attendance at religious services and importance of religious or spiritual expression. White blood cell counts, absolute numbers of lymphocytes, T-lymphocyte subsets, and natural killer cells were assessed by flow cytometry. Assessments of natural killer cell activity and delayed-type hypersensitivity responses to skin test antigens provided two measures of functional immunity. In analyses controlling for demographic, disease status, and treatment variables, women who rated spiritual expression as more important had greater numbers of circulating white blood cells and total lymphocyte counts. Upon examination of relationships with lymphocyte subsets, both helper and cytotoxic T-cell counts were greater among women reporting greater spirituality. PMID- 11906447 TI - Ductal eccrine carcinoma presenting as a Paget's disease-like lesion of the breast. AB - Some types of skin appendage tumors, particularly ductal eccrine carcinomas (DEC), imitate breast carcinomas histologically, thus causing great diagnostic challenges. We describe a case of DEC presenting as an eczematous, crusted skin lesion on the right nipple-areolar complex in a 67-year-old woman. A skin biopsy done under the clinical impression of Paget's disease of the nipple was initially misinterpreted as infiltrating ductal carcinoma, and a subsequent modified radical mastectomy revealed DEC exclusively confined to the nipple with perinodal tumor metastasis in one of the axillary lymph nodes. This case highlights the diagnostic difficulty caused by the histologic homology between breast carcinomas and skin appendage tumors with ductal differentiation. PMID- 11906446 TI - Lactating adenoma: a diagnosis of exclusion. AB - This is a case of a large lactating adenoma which developed in a 26 year old primagravida during the third trimester of her pregnancy. The presentation was that of a grossly enlarged and engorged breast with breast erythema, warmth and tenderness. Radiologic and pathologic examinations were difficult because of the lactational changes in the breast and areas of infarcted tissue within the large tumor. Skin biopsies and core biopsies of the mass were performed to exclude malignancy. Surgical resection of the mass was necessary for definitive diagnosis. The pathology proved to be a lactating adenoma, which is the most prevalent breast mass in young pregnant females. The large size of this tumor, and the presentation of breast erythema and edema raised the possibility of inflammatory breast cancer. Following surgical resection and definitive diagnosis of this tumor, the patient required plastic surgical reconstruction of the breast because of redundant breast tissue. Although most lactating adenomas spontaneously involute, the diagnosis is not always straight-forward and surgical resection may be required for definitive diagnosis and exclusion of other pathologic processes. PMID- 11906448 TI - Lipogranuloma. PMID- 11906449 TI - Mammographic evolution of fat necrosis of the breast. PMID- 11906450 TI - Linking fungal morphogenesis with virulence. AB - Pathogenic fungi have become an increasingly common cause of systemic disease in healthy people and those with impaired immune systems. Although a vast number of fungal species inhabit our planet, just a small number are pathogens, and one feature that links many of them is the ability to differentiate morphologically from mould to yeast, or yeast to mould. Morphological differentiation between yeast and mould forms has commanded attention for its putative impact on the pathogenesis of invasive fungal infections. This review explores the current body of evidence linking fungal morphogenesis and virulence. The topics addressed cover work on phase-locked fungal cells, expression of phase-specific virulence traits and modulation of host responses by fungal morphotypes. The effect of morphological differentiation on fungal interaction with host cells, immune modulation and the net consequence on pathogenesis of disease in animal model systems are considered. The evidence argues strongly that morphological differentiation plays a vital role in the pathogenesis of fungal infection, suggesting that factors associated with this conversion process represent promising therapeutic targets. PMID- 11906451 TI - Toxoplasma gondii Rab5 enhances cholesterol acquisition from host cells. AB - The role of endocytosis in nutrient uptake by Toxoplasma gondii is unknown. To explore this issue, we characterized an endosomal compartment by identifying a T. gondii Rab5 homologue, a molecular marker for early endosomes in eukaryotic cells. The deduced amino acid sequence of the T. gondii Rab5 gene encodes a protein of 240 amino acids, which we termed TgRab51. TgRab51 was epitope-tagged at the N-terminus, expressed in the parasite, and localized by immunofluorescence and immunoelectron microscopy to tubulovesicular structures anterior to the parasite nucleus and adjacent to, but distinct from the Golgi. By immunofluorescence analysis, TgRab51wt-HA staining partially overlapped with Golgi/TGN markers, but not with the T. gondii secretory organelles. A dominant positive mutant, TgRab51Q103L-HA, enhanced uptake of exogenous cholesterol analogues in intracellular parasites, augmented formation of lipid droplets and accelerated parasite growth. Brefeldin A disrupted the TgRab51 compartment, and altered the distribution of fluorescent exogenous cholesterol in cells expressing TgRab51Q103L-HA. These results suggest that TgRab51 facilitates sterol uptake, possibly through a Golgi-dependent pathway. PMID- 11906452 TI - The Salmonella pathogenicity island 1 secretion system directs cellular cholesterol redistribution during mammalian cell entry and intracellular trafficking. AB - The bacterial pathogen Salmonella triggers its own uptake into non-phagocytic mammalian cells. Entry is induced by the delivery of bacterial effector pro-teins that subvert signalling and promote cytoskeletal rearrangement, although the molecular mechanisms that co-ordinate initial pathogen-host cell recognition remain poorly characterized. Here we show that cholesterol is essential for Salmonella uptake. Depletion and chelation of plasma membrane cholesterol specifically inhibited bacterial internalization but not adherence. Cholesterol accumulated at bacterial entry sites in cultured cells, and was retained by Salmonella-containing vacuoles following pathogen internalization. Cellular cholesterol redistribution required bacterial effector protein delivery mediated by the Salmonella pathogenicity island (SPI) 1 type III secretion system, but was independent of the SPI2-encoded system. PMID- 11906453 TI - Association of a macrophage galactoside-binding protein with Mycobacterium containing phagosomes. AB - Mycobacteria reside intracellularly in a vacuole that allows it to circumvent the antimicrobial environment of the host macrophage. Although the mycobacterial phagosome exhibits selective fusion with vesicles of the endosomal system, identification of host and bacterial factors associated with phagosome bio genesis is limited. To identify these potential factors, mAbs were generated to a membrane preparation of mycobacterial phagosomes isolated from M. tuberculosis infected macrophages. A mAb recognizing a 32-35 kDa macrophage protein associated with the phagosomal membrane of Mycobacterium was identified. N-terminal sequence analysis identified this protein as Mac-2 or galectin-3, a galactoside-binding protein of macrophages. Galectin-3 (gal-3) was shown to accumulate in Mycobacterium-containing phagosomes during the course of infection. This accumulation was specific for phagosomes containing live mycobacteria and occurred primarily at the cytosolic face of the phagosome membrane. In addition, bind-ing of gal-3 to mycobacterial phosphatidylinositol mannosides (PIMs) demonstrated a novel interaction between host carbohydrate-binding proteins and released mycobacterial glycolipids. Infection of macrophages from gal-3-deficient mice indicated that the protein did not play a role in infection in vitro. In contrast, infection of gal-3-deficient mice revealed a reduced capacity to clear late but not early infection. PMID- 11906454 TI - Wild-type intracellular bacteria deliver DNA into mammalian cells. AB - Gene transfer in vitro from intracellular bacteria to mammalian phagocytic and non-phagocytic cells and in vivo in mice has been reported. The bacteria used as DNA delivery vectors were engineered to lyze upon entry in the cell due to impaired cell wall synthesis for Shigella flexneri and invasive Escherichia coli, or production of a phage lysin for Listeria mono- cytogenes. In vivo gene transfer was obtained with attenuated Salmonella typhimurium and resulted in stimulation of mucosal immunity. We report that wild-type intracellular human pathogens, such as L. monocytogenes EGD or LO28 and S. flexneri M90T, mediate efficient in vitro transfer of functional genes into epithelial and macrophage cell lines. A low- efficiency transfer was obtained from strain EGD to mouse peritoneal macrophages. DNA transfer with S. typhimurium was observed only from atten-uated aroA strain SL7207 into COS-1 cell line. As demonstrated by the study of listeriolysin-defective L. monocytogenes or of S. typhimurium SL7207 aroA engineered to secrete listeriolysin, escape of bacteria or of plasmid DNA from the intracytoplasmic vacuole is required for transfer of genetic information to occur. PMID- 11906455 TI - Microbial pathogenesis and the eukaryotic cytoskeleton at the American Society for Cell Biology annual meeting. PMID- 11906456 TI - Activation of alpha 1-adrenoceptors is not essential for the mediation of ischaemic preconditioning in rat heart. AB - 1. The aim of the present study was to clarify the role of alpha1-adrenoceptors in the mechanism of ischaemic preconditioning (IP). 2. Rat isolated perfused hearts were either non- preconditioned, preconditioned with 5 min ischaemia or treated for 5 min with alpha1-adrenoceptor agonists (50 micromol/L phenylephrine; 0.1, 0.5 and 1 micromol/L methoxamine) before being subjected to 45 min of sustained ischaemia followed by 60 min reperfusion. 3. Within each of the above protocols, hearts were divided into groups to which alpha1-adrenoceptor antagonists (prazosin, 5'-methyl urapidil and chloroethylclonidine (CEC)) were administered. Functional recovery and infarct size were used as indices of the effects of ischaemia. Ischaemic contracture characteristics and maximal diastolic pressure during reflow were also assessed. 4. Blockade of alpha(1)-adrenoceptors with prazosin or the subtype-selective antagonists 5'-methyl urapidil and CEC did not abolish the protective effect of IP with respect to both functional recovery and infarct size reduction. 5. Protection afforded by phenylephrine was attenuated in hearts treated with prazosin or the alpha(1B)-adrenoceptor- selective antagonist CEC, but not in those treated with the alpha(1A) adrenoceptor-selective antagonist 5'-methyl urapidil. 6. Treatment with low concentrations of methoxamine, considered to be alpha(1A)-adrenoceptor selective, did not confer any protection to the ischaemic myocardium. 7. A close relationship between accelerated ischaemic contracture and enhanced cardioprotection was observed. 8. The results suggest that alpha1-adrenoceptor stimulation mimics IP, but it is not an essential component in the mechanism behind the protective effect of IP in rat heart. In addition, the present study demonstrates that stimulation of the alpha(1B)- but not the alpha(1A) adrenoceptor subtype is responsible for the catecholamine-induced protection of ischaemic myocardium in rat. PMID- 11906457 TI - Distribution of Ca2+-activated K+ channel (SK2 and SK3) immunoreactivity in intestinal smooth muscles of the guinea-pig. AB - 1. The tissue distribution of small conductance Ca(2+)-activated K(+) channels (SK2 and SK3) was examined in three preparations of the guinea-pig intestine: the taneia caeci and the circular muscle layer of the stomach and proximal colon. 2. The SK3 immunoreactive (SK3-IR) cells were bi- or multipolar in appearance with numerous short processes, which formed an interconnecting network at the myenteric and submucous borders of the stomach and proximal colon. The SK3-IR cells were also present within the circular muscle layer of these preparations and throughout the taenia caeci. 3. Although SK3-IR cells had a similar distribution as cells immunoreactive for c-Kit (c-Kit-IR), the marker for interstitial cells of Cajal (ICC), only 5-10% of c-Kit-IR ICC were also SK3-IR. 4. The SK3-IR cells were clearly ICC when examined with the electron microscope. Close associations of SK3-IR ICC (ICC-SK3) and nerves were often observed, as were gap junctions with SK3-negative ICC and smooth muscle cells. 5. Punctate SK2 and SK3 channel immunoreactivity was present on the plasmalemmal surface of all smooth muscle cells examined. 6. We conclude that ICC-SK3 are a subpopulation of ICC that are directly innervated by enteric inhibitory motor nerves. PMID- 11906458 TI - Lack of impairment of nitric oxide-mediated responses in a rat model of high renin hypertension. AB - 1. Angiotensin (Ang) II triggers the expression of a pro- oxidant phenotype in the vascular wall, suggesting that activation of the renin-angiotensin system (RAS) causes endothelial dysfunction in various pathological situations, such as hypertension. However, this hypothesis has been mostly tested in a setting of exogenous administration of AngII. 2. We tested the hypothesis of a role for endogenous activation of the RAS leading to oxidant stress and endothelial dysfunction in a high-renin model of hypertension (i.e. two-kidney, one-clip hypertension) in rats. One month after clipping or sham surgery, aorta were isolated from untreated rats or rats treated by the angiotensin AT1 receptor antagonist irbesartan (10 mg/kg per day). Mesenteric artery segments were also isolated from normotensive or hypertensive rats. 3. Hypertension reduced the relaxations to acetylcholine but did not affect the ratio of contractions to phenylephrine in the presence compared with the absence of a nitric oxide (NO) synthase inhibitor, used as an index of basal release of NO. 4. The free radical scavenger tempol reduced the contractions to phenylephrine in the absence, but not in the presence, of an inhibitor of NO synthesis. This index of free radical mediated degradation of NO was not affected by hypertension. In parallel, hypertension did not affect the expression of p22phox, a component of the free radical generating enzyme reduced nicotinamide adenine dinucleotide phosphate oxidase. 5. Chronic treatment with the AT1 receptor antagonist decreased blood pressure, moderately improved the response to acetylcholine, but did not affect basal NO release in hypertensive rats, although it did increase basal NO release in normotensive rats. 6. Thus, this model of hypertension is characterized by an impaired stimulated NO release but not of basal NO release in isolated arteries. Furthermore, there was no functional evidence of an increased oxidative stress mediated impairment of NO release. This is not in favour of a direct link between activation of the RAS and development of endothelial dysfunction in experimental hypertension. PMID- 11906459 TI - L-arginine protects against stress-induced gastric mucosal lesions by preserving gastric mucus. AB - 1. We have shown that exogenously administered L-arginine protects against water immersion restraint (WIR) stress-induced gastric mucosal lesions in rats through preservation of nitric oxide (NO) generation via constitutive nitric oxide synthase (cNOS), but not inducible nitric oxide synthase (iNOS), in the gastric mucosa. We have also indicated that impaired gastric mucus synthesis and secretion occur through a decrease in gastric cNOS activity in WIR-stressed rats. Therefore, in the presesnt study, we examined whether exogenously administered L arginine exerts a protective effect against WIR stress-induced gastric mucosal lesions in rats through preservation of gastric mucus synthesis and secretion by NO generated from the administered amino acid via cNOS in the gastric mucosa. 2. Rats were subjected to WIR stress for 3 and 6 h. Either L-arginine (150-600 mg/kg) or D-arginine (600 mg/kg) was injected intraperitoneally 0.5 h prior to WIR stress. Either N(G)-monomethyl L-arginine (L-NMMA; 100 mg/kg) or N(G) monomethyl D-arginine (D-NMMA; 100 mg/kg) was injected subcutaneously 0.5 h prior to WIR stress. Total NOS, cNOS, iNOS, nitrite and nitrate (breakdown products of NO), hexosamine (an index of gastric mucin) and adherent mucus were assayed in the gastric mucosa. 3. Pretreatment with L-arginine, but not D-arginine, protected against gastric mucosal lesions in rats subjected to WIR stress for 3 and 6 h in a dose-dependent manner. Pretreatment with L-arginine, but not D arginine, attenuated decreases in hexosamine and adherent mucus concentrations and cNOS activity and increases in total NOS and iNOS activities and nitrite/nitrate concentration in the gastric mucosal tissue of rats subjected to WIR stress for 3 and 6 h in a dose-dependent manner. Both the protective effect of L-arginine against gastric mucosal lesions and the attenuating effect of the amino acid on the decreases in gastric mucosal hexosamine and adherent mucus concentrations and cNOS activity in rats subjected to WIR stress for 6 h were counteracted by cotreatment with L-NMMA, a nitric oxide synthase inhibitor, but not D-NMMA. 4. These results suggest that exogenously administered L-arginine exerts a protective effect against stress-induced gastric mucosal lesions in rats at least partly through preservation of gastric mucus synthesis and secretion by NO produced from the administered amino acid via cNOS in gastric mucosal tissue. PMID- 11906460 TI - Uneven changes in circulating blood cell counts with adrenergic stimulation to the canine spleen. AB - 1. Responses of splenic diameter measured by sonomicrometry to alpha- and beta adrenoceptor stimulants were estimated together with simultaneously measured systemic arterial and splenic venous concentrations of red blood cells (RBC), white blood cells (WBC) and platelets (PLT) in anaesthetized dogs. 2. Intravenous and intrasplenic arterial injections of adrenaline, noradrenaline and phenylephrine all produced profound decreases in splenic diameter. Increases in systemic arterial concentrations of RBC produced by these stimulants were observed immediately following the splenic contraction and marked increases in splenic venous concentration of RBC. 3. Adrenaline and noradrenaline both caused rapid and transient decreases in WBC release from the spleen and these agonists then, in addition to phenylephrine, caused gradual and small increases in systemic arterial WBC concentrations. The increases in arterial RBC and WBC concentrations were abolished by transient isolation of the spleen from the systemic circulation. These three agents did not significantly modify differences between splenic venous and systemic arterial concentrations of PLT. 4. Intra arterial injections of isoprenaline caused an increase in splenic diameter and a significant decrease in the splenic venous concentration of WBC, but barely influenced the veno-arterial differences in RBC and PLT concentrations across the spleen. 5. The present results indicate that contraction of the spleen through stimulation of splenic alpha1-adrenoceptors contributes to supplying RBC but not PLT into the systemic circulation, whereas splenic beta-adrenoceptor activation leads to splenic dilatation with increased sequestration of WBC to the spleen despite no changes in the release of RBC and PLT from the spleen. PMID- 11906461 TI - Effect of various oestrogens on cell injury and alteration of apical transporters induced by tert-butyl hydroperoxide in renal proximal tubule cells. AB - 1. The present study was undertaken in order to examine the effect of various oestrogens on tert-butyl hydroperoxide (t-BHP)-induced cell injury and changes in apical transporters in primary cultured rabbit renal proximal tubule cells. 2. Compared with control, t-BHP (0.5 mmol/L; 1 h) decreased cell viability (62%) and glutathione (GSH) content (60%) and increased lipid peroxide (LPO) formation (309%), arachidonic acid (AA) release (193%) and Ca(2+) influx (168%). 3. The protective potency of various oestrogens for these parameters is dependent on the precise oestrogenic structure, with 2-hydroxy-oestradiol-17 beta (2-OH-E(2)) and 4-OH-E(2), both catecholic oestrogens, or diethylstilbesterol (DES) being more potent than oestradiol (E(2)), oestriol or oestradiol-17 alpha, all phenolic oestrogens (P < 0.05). 4. These cytoprotective effects of oestrogens occur at concentrations above 10 micromol/L and are not dependent on classical oestrogen receptors and gene transcription and translation. In addition, various oestrogens have different preventative effects against t-BHP-induced inhibition of [(14)C] alpha-methyl-D-glucopyranoside (alpha-MG), inorganic phosphate (Pi) and Na(+) uptake, consistent with the results of cell injury. In contrast, the potency against t-BHP-induced changes in cell viability, LPO, GSH content and transporter function of the anti-oxidants taurine and vitamin C is similar to that of phenolic oestrogens, whereas that of the iron chelators deferoxamine and phenanthroline is similar to that of catecholic oestrogens. 5. In conclusion, various oestrogens have differential cytoprotective potential against t-BHP induced cell injury and decreases in alpha-MG, Na(+) and Pi uptake. These effects are due, in part, to both the basic chemical properties of the compounds and the maintenance of endogenous GSH or inhibition of AA release and Ca(2+) influx. PMID- 11906462 TI - Cerebral thrombosis and microcirculation of the rat during the oestrous cycle and after ovariectomy. AB - 1. The effects of oestrogen on thrombogenesis and the cerebral microcirculation of the female rat were studied during the oestrous cycle and after ovariectomy. 2. Serum levels of oestradiol (E2) and plasma concentrations of nitric oxide (NO) metabolites were significantly greater at pro-oestrus than at dioestrus. Blood vessel diameter, mean red cell velocity, wall shear rate and blood flow at pro oestrus were significantly higher than at dioestrus. Thrombotic tendency, assessed using a He-Ne laser-induced thrombosis model, was significantly decreased at pro-oestrus compared with dioestrus. 3. The long-term deprivation of oestrogen by ovariectomy significantly depressed serum levels of E2 and plasma concentrations of NO metabolites. Thrombotic tendency was significantly increased 4 weeks after ovariectomy. Vessel diameter, mean red cell velocity, wall shear rate and blood flow in pial arterioles were significantly reduced after ovariectomy. 4. Exogenous administration of oestrogen (17 beta-oestradiol) after surgery reversed the increased thrombotic tendency mediated by ovariectomy. 5. These results strongly indicate that oestrogen mediates beneficial effects on the cerebral microcirculation and moderates cerebral thrombotic mechanisms in the female rat. PMID- 11906463 TI - Concentration of egg white lysozyme in the serum of healthy subjects after oral administration. AB - 1. While the egg white lysozyme preparation ER0068 (Neuzym; Eisai, Tokyo, Japan) is widely used clinically, no studies have been performed on its pharmacokinetic properties at clinically relevant doses. In the present study, we used a highly sensitive two-site enzyme immunoassay in order to determine the pharmacokinetic properties of egg white lysozyme after oral administration of two doses within the clinical range, paying particular attention to the effects of food intake. 2. A total of 22 healthy male subjects aged 20-45 years participated in the study. All subjects had been screened for egg white allergy and non-specific lysozyme inhibitors in their serum. Subjects who received 90 mg ER0068 after an overnight fast reached a maximum serum concentration of 1700 pg/mL within 1 h, compared with non-detectable levels in untreated controls. In a second experiment, subjects received 30 and 90 mg ER0068 after an overnight fast and 90 mg in the non-fasted state and exhibited maximum serum levels of 37, 360 and 49 pg/mL, respectively. Egg white lysozyme concentrations in serum returned to undetectable levels after a maximum of 48 h. 3. We conclude that clinically relevant concentrations of egg white lysozyme are absorbed in significant amounts, despite its high molecular weight. However, food intake considerably reduces the amount of enzyme absorbed. PMID- 11906464 TI - Garlic and its active metabolite allicin produce endothelium- and nitric oxide dependent relaxation in rat pulmonary arteries. AB - 1. The aims of the present study were to investigate the effects of fresh garlic and one of its active metabolites, allicin, on rat isolated pulmonary arteries (RPA). 2. In endothelium-intact and phenylephrine-precontracted RPA, the addition of a water or a 5% ethanol extract of fresh garlic (1-500 microg/mL) resulted in a dose-dependent relaxation reaching a maximum (mean +/- SEM) of -91 +/- 3 and 93 +/- 2%, respectively, with an ED(50) of 113 +/- 12 and 106 +/- 10 microg/mL, respectively. The vasorelaxation was readily reversible upon washing and no tachyphylaxis was noted. 3. An extract of the external garlic storage leaf produced a significantly greater relaxation than the inner stem. Microfiltration of extracts with a 10,000 molecular sieve did not attenuate relaxation. Inactivation of alliinase and allicin formation, with either boiling of the garlic clove for 30 min or 100% ethanol treatment, completely abolished relaxation. In contrast, similar treatment of crushed garlic with formed allicin retained the relaxation response. 4. Pure allicin produced a similar relaxation as garlic extract, with an EC(50) of approximately 0.8 microg/mL. Disruption of endothelium or N(G)-nitro-L-arginine methyl ester pretreatment attenuated the relaxation, whereas indomethacin had no effect. 5. Prior garlic (500 microg/mL) treatment enhanced acetylcholine relaxation by shifting the response curve to the left, but had no effect on nitric oxide (NO) donor-induced responses. 6. These results demonstrate that garlic and the active metabolite allicin are capable of eliciting a NO-dependent relaxation in RPA and that this response is likely to be mediated via garlic activation of NO formation rather than its stabilization. PMID- 11906465 TI - Nitric oxide-mediated vasodilatory effect of atrial natriuretic peptide in forearm vessels of healthy humans. AB - 1. The aim of the present study was to determine whether the vasorelaxant effect of atrial natriuretic peptide (ANP) is, in part, endothelium dependent in humans. 2. We used veno-occlusive plethysmography to measure forearm blood flow (FBF) during intra-arterial infusions of ANP (4, 8, 16, 32 pmol/min per dL forearm tissue volume) before and after the inhibition of nitric oxide (NO) synthesis by N(G)-monomethyl-L-arginine (L-NMMA; 100 micromol) in seven normal healthy subjects. 3. Atrial natriuretic peptide caused a dose-dependent increase in FBF both before and after L-NMMA and significantly reduced the plasma concentration of angiotensin (Ang) II. Administration of L-NMMA significantly diminished the increase in FBF in response to ANP infusion (P < 0.05). 4. These results suggest that the forearm vasodilative response to ANP is modulated, in part, by an endothelium-derived NO-mediated mechanism associated with a decrease in AngII caused by ANP. PMID- 11906466 TI - The vestibulosympathetic reflex in humans: neural interactions between cardiovascular reflexes. AB - 1. Over the past 5 years, there has been emerging evidence that the vestibular system regulates sympathetic nerve activity in humans. We have studied this issue in humans by using head-down rotation (HDR) in the prone position. 2. These studies have clearly demonstrated increases in muscle sympathetic nerve activity (MSNA) and calf vascular resistance during HDR. These responses are mediated by engagement of the otolith organs and not the semicircular canals. 3. However, differential activation of sympathetic nerve activity has been observed during HDR. Unlike MSNA, skin sympathetic nerve activity does not increase with HDR. 4. Examination of the vestibulosympathetic reflex with other cardiovascular reflexes (i.e. barorereflexes and skeletal muscle reflexes) has shown an additive interaction for MSNA. 5. The additive interaction between the baroreflexes and vestibulosympathetic reflex suggests that the vestibular system may assist in defending against orthostatic challenges in humans by elevating MSNA beyond that of the baroreflexes. 6. In addition, the further increase in MSNA via otolith stimulation during isometric handgrip, when arterial pressure is elevated markedly, indicates that the vestibulosympathetic reflex is a powerful activator of MSNA and may contribute to blood pressure and flow regulation during dynamic exercise. 7. Future studies will help evaluate the importance of the vestibulosympathetic reflex in clinical conditions associated with orthostatic hypotension. PMID- 11906467 TI - Neural circuits controlling cardiorespiratory responses: baroreceptor and somatic afferents in the nucleus tractus solitarius. AB - 1. A vast array of peripheral receptors provide the central nervous system (CNS) with sensory signals that coordinate autonomic motor outflow to cardiovascular organs, such as the heart and peripheral vasculature, during locomotion. 2. Much of this sensory input is mediated by cardiovascular receptors located in blood vessels (arterial baroreceptors) and skeletal muscle (skeletal muscle ergoreceptors). 3. Several medullary nuclei are targets for cardiovascular receptors, including the nucleus tractus solitarius (NTS). 4. In the present review, the interaction between arterial baroreceptor and somatosensory receptor afferents in the NTS is examined while placing particular emphasis on the neurochemical and electrophysiological mechanisms involved in processing these signals. 5. Data from anaesthetized animals, as well as from an innovative working heart-brainstem preparation, will illustrate the potential role of GABAergic transmission on baroreceptor signalling in the caudal NTS during locomotion. PMID- 11906468 TI - Role of the vestibular system in regulating respiratory muscle activity during movement. AB - 1. Changes in posture can affect the resting length of the diaphragm, which is corrected through increases in both diaphragm and abdominal muscle activity. Furthermore, postural alterations can diminish airway patency, which must be compensated for through increases in firing of particular upper airway muscles. 2. Recent evidence has shown that the vestibular system participates in adjusting the activity of both upper airway muscles and respiratory pump muscles during movement and changes in body position. 3. Vestibulo-respiratory responses do not appear to be mediated through the brainstem respiratory groups; labyrinthine influences on respiratory pump muscles may be relayed through neurons in the medial medullary reticular formation, which have recently been demonstrated to provide inputs to both abdominal and diaphragm motoneurons. 4. Three regions of the cerebellum that receive vestibular inputs, the fastigial nucleus, the nodulus/uvula and the anterior lobe, also influence respiratory muscle activity, although the physiological role of cerebellar regulation of respiratory activity is yet to be determined. 5. It is practical for the vestibular system to participate in the control of respiration, to provide for rapid adjustments in ventilation such that the oxygen demands of the body are continually matched during movement and exercise. PMID- 11906469 TI - Balancing acts: respiratory sensations, motor control and human posture. AB - 1. The present brief review covers some novel aspects of integration between respiration and movement of the body. 2. There are potent viscerosomatic reflexes in animals involving small-diameter pulmonary afferents that, when excited, would limit exercise. However, recent studies using lobeline injections to excite pulmonary afferents in awake humans suggest that there is no evoked reflex motoneuronal inhibition. Instead, the noxious respiratory sensations generated by the vagal afferents may be crucial in the decision to stop exercise. 3. While respiratory movements may affect limb movements, the control of the trunk and limbs can involve interaction (and even interference) with key respiratory muscles, such as the diaphragm. Recent studies have revealed that not only does the diaphragm receive feed-forward drive prior to some limb movements, but that it also contracts both phasically and tonically during repetitive limb movements. 4. Thus, challenges to posture can indirectly challenge ventilation, while coordinated diaphragm contraction may contribute to control of the trunk. PMID- 11906470 TI - Exercise and hypertension: a model for central neural plasticity. AB - 1. Physical movement is accompanied by coordinated changes in respiratory and cardiovascular activity proportional to the metabolic demands of the locomotor task. Cardiorespiratory changes include increases in ventilation, blood pressure and heart rate, as well as altered regional sympathetic nerve activity and blood flow. 2. The posterior hypothalamic area, a periventricular region in the caudal most diencephalon, has been shown to play a role in mediating the coupling of locomotion and cardiorespiratory activity. Stimulation of this brain region produces locomotor behaviour and simultaneous increases in cardiorespiratory activity that are independent of peripheral feedback from contracting muscles. Posterior hypothalamic neurons are also activated by exercise and exercise related stimuli, such as muscle contraction. 3. In spontaneously hypertensive rats (SHR), a deficiency in the inhibitory GABA neurotransmitter system within the posterior hypothalamic area contributes to tonically elevated levels of arterial blood pressure. We previously identified a reduction in the GABA synthesizing enzyme glutamic acid decarboxylase (GAD) within the posterior hypothalamus of SHR. 4. We have recently demonstrated that exercise can upregulate GABA-mediated caudal hypothalamic control of cardiovascular function in SHR. Similarly, exercise increases GAD gene transcript levels in the posterior hypothalamus. Thus, we have identified a model to study exercise-related central neural plasticity in GABAergic neurotransmitter function. Moreover, we suggest that exercise may increase cardiovascular health through changing central neural regulation of blood pressure. PMID- 11906471 TI - Blood pressure control early in diabetes: a balance between angiotensin II and nitric oxide. AB - 1. Hyperglycaemia can lead to hypertension in long- standing diabetes through cumulative effects to cause progressive glomerular injury. However, it is not known how hyperglycaemia directly affects blood pressure control. Moreover, it has been difficult to isolate the actions of hyperglycaemia from other factors that can influence blood pressure. Recent studies at the earliest stages of type I diabetes have started to address these issues. 2. In humans and in animal models of type I diabetes, there is evidence that onset of hyperglycaemia can increase blood pressure. The increase is mild and has been linked to angiotensin (Ang) II, possibly due to slight overstimulation of the system in response to hyperglycaemia. 3. We have reported recently that onset of hyperglycaemia causes severe hypertension in rats that are treated chronically with N(G)-nitro-L arginine methyl ester and the hypertension is accompanied by a progressive increase in AngII and absence of hyperfiltration. This suggests that nitric oxide (NO), at the early stages of diabetes, is important in mediating the increase in glomerular filtration rate and in preventing hypertension. 4. We propose a hypothesis based on these results that hyperglycaemia at the earliest stages of diabetes induces hyperfiltration and natriuresis and also activates a compensatory hypertensive stimulus (likely AngII) in an attempt to maintain normal blood pressure. A critical balance exists between these forces and NO plays a pivotal role by attenuating AngII- mediated renal vasoconstriction and facilitating the increase in glomerular filtration rate. PMID- 11906472 TI - A metabolic mechanism for cardiac K+ channel remodelling. AB - 1. Electrical remodelling of the ventricle is a common pathogenic feature of cardiovascular disease states that lead to heart failure. Experimental data suggest this change in electrophysiological phenotype is largely due to downregulation of K(+) channels involved in repolarization of the action potential. 2. Voltage-clamp studies of the transient outward current (I(to)) in diabetic cardiomyopathy support a metabolic mechanism for K(+) channel downregulation. In particular, I(to) density is significantly increased in diabetic rat isolated ventricular myocytes treated in vitro with insulin or agents that activate pyruvate dehydrogenase. Recent data suggest this mechanism is not limited to diabetic conditions, because metabolic stimuli that upregulate I(to) in diabetic rat myocytes act similarly in non- diabetic models of heart failure. 3. Depressed I(to) channel activity is also reversed by experimental conditions that increase myocyte levels of reduced glutathione, indicating that oxidative stress is involved in electrical remodelling. Moreover, upregulation of I(to) density by activators of glucose utilization is blocked by inhibitors of glutathione metabolism, supporting the premise that there is a functional link between glucose utilization and the glutathione system. 4. Electrophysiological studies of diabetic and non-diabetic disease conditions affecting the heart suggest I(to) channels are regulated by a redox-sensitive mechanism, where glucose utilization plays an essential role in maintaining a normally reduced state of the myocyte. This hypothesis has implications for clinical approaches aimed at reversing pathogenic electrical remodelling in a variety of cardiovascular disease states. PMID- 11906473 TI - Multiple mechanisms of early hyperglycaemic injury of the rat intestinal microcirculation. AB - 1. Hyperglycaemia in the vast majority of humans with diabetes mellitus is the end result of profound insulin resistance secondary to obesity. For patients in treatment, hyperglycaemia is usually not sustained but, rather, occurs intermittently. In in vivo studies of the rat intestinal microcirculation, endothelial impairment occurs within 30 min at D-glucose concentrations > or = 300 mg/dL. Endothelial-dependent dilation to acetylcholine and constriction to noradrenaline is impaired. Vasodilation to exogenous nitric oxide (NO) remains normal. 2. When initiated before hyperglycaemia, suppression of oxygen radicals by both scavenging and pretreatment with cyclo-oxygenase blockade to prevent oxygen radical formation minimized endothelial impairments during hyperglycaemia. Neither treatment was effective in restoring endothelial function once it was damaged by hyperglycaemia. 3. A mechanism that may initiate the arachidonic acid- oxygen radical process is activation of specific isoforms of protein kinase C (PKC). De novo formation of diacylglycerol during hyperglycaemia activates PKC. Blockade of the beta II PKC isoform with LY-333531 prior to hyperglycaemia protected NO formation within the arteriolar wall, as judged with NO-sensitive microelectrodes. Furthermore, once suppression of endothelial dilation was present in untreated animals, PKC blockade could substantially restore endothelial-dependent dilation. 4. These results indicate that acute hyperglycaemia is far from benign and, in the rat, causes rapid endothelial impairment. Both oxygen radical scavenging and cyclo-oxygenase blockade prior to bouts of hyperglycaemia minimize endothelial impairment with limited side effects. Blockade of specific PKC isozymes protects endothelial function both as a pre- or post-treatment during moderately severe hyperglycaemia. PMID- 11906475 TI - Potentiation of glucose-mediated glomerular injury by mechanical strain. AB - 1. The glomerular injury of diabetes is characterized by the progressive accumulation of extracellular matrix in the mesangial regions, ultimately resulting in glomerulosclerosis. 2. The excessive glomerular extracellular matrix formation associated with the haemodynamic alteration of diabetes is the result of mesangial mechanical strain. 3. The increased synthesis and deposition of extracellular matrix is augmented by the presence of high glucose concentrations. 4. Both mechanical strain and high glucose share many of the mechanisms mediating their metabolic effects, including the stimulation of prosclerotic growth factors. 5. Little is known about factors that may influence the long-term effects of mechanical strain, but the preservation of the F-actin cytoskeleton is likely an important modulator of the resulting injury. PMID- 11906474 TI - Altered electromechanical coupling in the renal microvasculature during the early stage of diabetes mellitus. AB - 1. The early stage of type 1 diabetes mellitus (DM) is characterized by renal hyperfiltration, which promotes the eventual development of diabetic nephropathy. The hyperfiltration state is associated with afferent arteriolar dilation and diminished responsiveness of this vascular segment to a variety of vasoconstrictor stimuli, whereas efferent arteriolar diameter and vasoconstrictor responsiveness are typically unaltered. 2. The contractile status of preglomerular vascular smooth muscle appears to be tightly coupled to membrane potential (E(m)) and its influence on Ca(2+) influx through voltage-gated channels. Efferent arteriolar tone is largely independent of electromechanical events. Hence, defective electromechanical mechanisms in vascular smooth muscle should engender selective changes in preglomerular microvascular function, such as those evident during the early stage of DM. 3. Afferent arteriolar contractile responses to K(+)-induced depolarization and BAYK8644 are diminished 2 weeks after onset of DM in the rat. Similarly, depolarization-induced Ca(2+) influx and the resulting increase in intracellular [Ca(2+)] are abated in the preglomerular microvasculature of diabetic rats. The intracellular [Ca(2+)] response to depolarization is rapidly restored by normalization of extracellular glucose levels. These observations suggest that hyperglycaemia in DM impairs regulation of afferent arteriolar voltage-gated Ca(2+) channels. 4. Dysregulation of E(m) may also contribute to afferent arteriolar dilation in DM. Vasodilator responses to pharmacological opening of ATP-sensitive K(+) channels are exaggerated in afferent arterioles from diabetic rats. Moreover, blockade of these channels normalizes afferent arteriolar diameter in kidneys from diabetic rats. These observations suggest that increased functional availability and basal activation of ATP-sensitive K(+) channels promote afferent arteriolar dilation in DM. 5. We propose that dysregulation of E(m) (involving ATP- sensitive K(+) channels) and a diminished Ca(2+) influx response to depolarization (involving voltage-gated Ca(2+) channels) may act synergistically to promote preglomerular vasodilation during the early stage of DM. PMID- 11906476 TI - Endothelium-derived hyperpolarizing factor: is there a novel chemical mediator? AB - 1. Endothelium-derived hyperpolarization (EDH) has been reported in many vessels and an extensive literature suggests that a novel, non-nitric oxide and non prostanoid, endothelium-derived factor(s) may be synthesized in endothelial cells. 2. The endothelium-dependent hyperpolarizing factor, or EDHF, is synthesized by the putative EDHF synthase and mediates its cellular effects by either, directly or indirectly, opening K channels on vascular smooth muscle cells or, via hyperpolarization of the endothelial cell, by facilitating electrical coupling between the endothelial and the vascular smooth muscle cell. 3. The question of the chemical identity of EDHF has received considerable attention; however, no consensus has been reached. Tissue and species heterogeneity exists that may imply there are multiple EDHF. Leading candidate molecules for EDHF include an arachidonic acid product, possibly an epoxygenase product, or an endogenous cannabinoid, or simply an increase in extracellular K+. 4. An increasing body of evidence suggests that EDH, notably in the resistance vasculature, may be mediated via electrical coupling through myoendothelial gap junctions and the existence of electrical coupling may negate the need to hypothesize the existence of a true endothelium-derived chemical mediator. 5. In this paper we review the evidence that supports and refutes the existence of a novel EDHF versus a hyperpolarization event mediated solely by myoendothelial gap junctions. PMID- 11906477 TI - Heterogeneity of the properties of INa in epicardial and endocardial cells of rat ventricle. AB - 1. Ventricular I(Na) heterogeneity was investigated in adult rat hearts. Differences in transient outward potassium current (Ito) were used to confirm isolation of subepicardial and subendocardial cells. Mean peak Ito was 6.0 +/- 0.7 and 1.6 +/- 0.45 pA/pF in epicardial and endocardial cells, respectively (P < < 0.01). 2. Maximum sodium conductance was smaller in subendocardial cells compared with subepicardial cells (2.39 +/- 0.11 vs 2.78 +/- 0.12 nS/pF, respectively; n=17 for both; 0.01 < P < 0.05) and 50% activation occurred at a slightly more negative potential (-47.6 +/- 0.8 vs -44.9 +/- 0.9 mV, respectively; n=10 for both; 0.01 < P < 0.05). 3. The potential for 50% inactivation was not significantly different in subepicardial compared with subendocardial cells (72.2 +/- 1.0 vs 72.8 +/- 2.2 mV, respectively; n=17 for both; NS). 4. Persistent sodium current density appeared smaller in subendocardial (n =19) compared with subepicardial (n=11) cells (at a test potential of -25 mV current, density was 0.118 +/- 0.041 vs 0.144 +/- 0.085 pA/pF, respectively), although this was not statistically significant due to large variability between cells. 5. Mathematical modelling of the cardiac action potential indicated that the combined effects of differences in current density and voltage dependence of sodium currents are unlikely to contribute to ventricular action potential heterogeneity between epicardial and endocardial cells. PMID- 11906478 TI - Possible involvement of P-glycoprotein in the biliary excretion of grepafloxacin. AB - 1. In the present study, we have examined the effects of the quinolones norfloxacin (NFLX), enoxacin (ENX), ofloxacin (OFLX), tosufloxacin (TFLX), lomefloxacin (LFLX), sparfloxacin (SPFX) and grepafloxacin (GPFX) on the efflux of doxorubicin from mouse leukaemia P388/ADR cells expressing P-glycoprotein. The relationship between their partition coefficients (hydrophobicity) and effluxing potencies was also elucidated. 2. Both TFLX and SPFX strongly increased the intracellular accumulation of doxorubicin (5 micromol/L) in P388/ADR cells, but had no effect on P388/S cells not expressing P-glycoprotein. The rank of order of the potency of the quinolones (TFLX > SPFX > GPFX > NFLX) was not related directly to their hydrophobicity. These results suggest that some quinolones can reverse anticancer drug resistance. 3. Because GPFX is more highly excreted into the bile than other known quinolones, the effects of doxorubicin (10 mg/kg) or the well-known inhibitors of P-glycoprotein, namely cyclosporine A (10 mg/kg) and erythromycin (100 mg/kg), on the biliary excretion of GPFX at steady state was studied in rats. 4. Doxorubicin, cyclosporine A and erythromycin significantly decreased the biliary clearance of GPFX. Cyclosporine A and erythromycin had a much stronger inhibitory effect on the biliary excretion of GPFX than doxorubicin. These results suggest the possibility that GPFX is, at least in part, excreted into the bile by a P-glycoprotein-mediated transport mechanism. PMID- 11906480 TI - Characterization of lymphocyte beta 2-adrenoceptor signalling in patients with left ventricular volume overload disease. AB - 1. Studies using animal experimental models have suggested that the beta2 adrenoceptor is uncoupled in association with alterations in the expression of G protein-coupled receptor kinases (GRK) 2/3 in heart failure. However, the functional expression of the components of this pathway in human disease has not been fully elucidated yet. In the present study, we evaluated the possibility that the regulation of beta2-adrenoceptor signalling components in patients with left ventricular volume overload (VOL) depends on the severity of the overload. 2. We characterized the lymphocyte GRK 2-6, beta-arrestins 1 and 2, beta2 adrenoceptor expression at the mRNA and protein levels, as well as the activity of adenylyl cyclase, protein kinases (PK) A and PKC in patients with VOL using healthy blood donors as controls. 3. In the patient group, GRK2 mRNA was increased by 61% (P < 0.001), GRK3 was increased by 54% (P < 0.005), GRK5 was increased fivefold (P < 0.001) and the beta-arrestin 2 mRNA was increased by 40% (P < 0.05). These increases were paralleled with a sixfold increase in GRK2, a twofold increase in GRK3 and a 1.3-fold increase in GRK5 protein levels. These changes were associated with a significant decrease in beta2-adrenoceptor mRNA, the basal, catalytic and receptor-mediated activity of adenylyl cyclase and sensitization of the forskolin-stimulated activity towards augmented inhibition by guanylimidodiphosphate. In general, the increase in GRK2 and 5 mRNA exhibited a positive correlation with the gravity of the haemodynamic load, as determined by changes in left ventricular fractional shortening. 4. The results suggest that VOL induces an increase in the expression of lymphocyte beta2-adrenoceptor specific GRK and beta-arrestin 2 in association with an attenuation in beta2 adrenoceptor levels. It can be speculated that the cardiac circulatory system adapts itself to altered haemodynamic functional demands partly by altering beta2 adrenoceptor signalling. PMID- 11906479 TI - A comparative study on the activity of lansoprazole, omeprazole and PD-136450 on acidified ethanol- and indomethacin-induced gastric lesions in the rat. AB - 1. The proton pump inhibitors lansoprazole (LP) and omeprazole (OP) and the cholecystokinin (CCK)-receptor antagonist PD-136450 (PD) provide a broad spectrum of activities in their ability to inhibit gastric acid secretion and protect the stomach against ulcerogens. In the present study, we investigated the protective effects of these compounds against gastric ulcers induced by acidified ethanol (AE) and indomethacin. 2. Both AE (60% ethanol in 150 mmol/L HCl, 1 mL/rat) and indomethacin (30 mg/kg) produced gastric haemorrhagic lesions in the rat 1 and 6 h after oral administration, respectively. 3. The gastric mucosal protective effects of LP (1-20 mg/kg), OP (0.5-10 mg/kg) and PD (1-20 mg/kg), administered either orally or subcutaneously (s.c.) 30 min before the administration of AE or indomethacin, were dose dependent against both models of ulcer induction. 4. To determine whether the cytoprotective effect of LP, OP and PD (each 10 mg/kg) was mediated by endogenous prostaglandins (PG), indomethacin (10 mg/kg, s.c.) was administered 15 min before AE to inhibit prostanoids biosynthesis. Indomethacin reduced the cytoprotective effects of OP, but not LP, administered either orally or s.c. Indomethacin reduced the cytoprotective effect of PD administered orally, although the effect was much less significant than when PD was administered s.c. The results exclude the role of PG in mediating the protective effects of LP, whereas the possibility exists for PG to have a role in mediating the protective effects of OP and PD. 5. To investigate the possible involvement of endogenous nitric oxide (NO) in the cytoprotective action of LP, OP and PD, we treated rats with a selective inhibitor of NO synthesis, namely NG-nitro-L-arginine methyl ester (L-NAME; 25 mg/kg, s.c.). Administration of L-NAME 15 min prior to LP, OP or PD (each 10 mg/kg) orally or s.c. and challenge with AE or indomethacin did not significantly increase the degree of the ulcer index and L-NAME was not able to antagonize the protective effects of LP, OP and PD, thus excluding the role of NO in mediating the protective effects of these drugs. However, the effects of PD in reducing the indomethacin-induced ulcer index were less significant in the presence than the absence of L-NAME (P < 0.05 vs P < 0.001, respectively), suggesting a role for NO. 6. In conclusion, the results of the present study suggest that LP and OP are equally effective against AE- as well as indomethacin induced gastric ulcers and were more potent than PD in protecting the stomach against ulcer formation. Lansoprazole, OP and PD bring about their cytoprotective action through the reduction of acid secretion and some other unknown mechanisms. However, OP and PD may exert their cytoprotective action through PG and NO pathways. PMID- 11906481 TI - Protective effect of alpha-lipoic acid against ischaemic acute renal failure in rats. AB - 1. In the present study, we investigated whether treatment with alpha-lipoic acid (LA), a powerful and universal anti-oxidant, has renal protective effects in rats with ischaemic acute renal failure (ARF). 2. Ischaemic ARF was induced by occlusion of the left renal artery and vein for 45 min followed by reperfusion, 2 weeks after contralateral nephrectomy. Blood urea nitrogen (BUN), plasma concentrations of creatinine (Pcr) and urinary osmolality (Uosm) were measured for the assessment of renal dysfunction. Creatinine clearance (Ccr) and fractional excretion of Na+ (FENa) were used as indicators of glomerular and tubular function, respectively. 3. Renal function in ARF rats decreased markedly 24 h after reperfusion. Intraperitoneal injection of LA at a dose of 10 mg/kg before the occlusion tended to attenuate the deterioration of renal function. A higher dose of LA (100 mg/kg) significantly (P < 0.01) attenuated the ischaemia/reperfusion-induced increases in BUN (19.1 +/- 0.7 vs 7.2 +/- 0.7 mmol/L before and after treatment, respectively), Pcr (290 +/- 36 vs 78.1 +/- 4.2 micromol/L before and after treatment, respectively) and FENa (1.39 +/- 0.3 vs 0.33 +/- 0.09% before and after treatment, respectively). Treatment with 100 mg/kg LA significantly (P < 0.01) increased Ccr (0.70 +/- 0.13 vs 2.98 +/- 0.27 mL/min per kg before and after treatment, respectively) and Uosm (474 +/- 39 vs 1096 +/- 80 mOsmol/kg before and after treatment, respectively). 4. Histopathological examination of the kidney of ARF rats revealed severe lesions. Tubular necrosis (P < 0.01), proteinaceous casts in tubuli (P < 0.01) and medullary congestion (P < 0.05) were significantly suppressed by the higher dose of LA. 5. A marked increase in endothelin (ET)-1 content in the kidney after ischaemia/reperfusion was evident in ARF rats (0.43 +/- 0.02 ng/g tissue) compared with findings in sham- operated rats (0.20 +/- 0.01 ng/g tissue). Significant attenuation (P < 0.01) of this increase occurred in ARF rats treated with the higher dose of LA (0.24 +/- 0.03 ng/g tissue). 6. These results suggest that administration of LA to rats prior to development of ischaemic ARF prevents renal dysfunction and tissue injury, possibly through the suppression of overproduction of ET-1 in the postischaemic kidney. PMID- 11906482 TI - Dendroaspis natriuretic peptide administered intracerebroventricularly increases renal water excretion. AB - 1. The effects of intracerebroventricular (i.c.v.) infusion of Dendroaspis natriuretic peptide (DNP) on renal function were examined in anaesthetized rats. The results were compared with those obtained following intravenous (i.v.) infusion of the same dose of DNP. 2. Urine volume was increased four- to six-fold over basal values by i.c.v. infusion of DNP (6 pmol/min). Urine osmolality was decreased, whereas sodium excretion was not significantly altered. Intravenous infusion of the same dose of DNP had little effect on urinary water excretion. Neither arterial pressure nor heart rate was changed significantly by either i.v. or i.c.v. infusion of DNP. Glomerular filtration rate, measured by creatinine clearance, remained unaltered. 3. The diuretic response to i.c.v. DNP was markedly attenuated in rats that were deprived of their water supply for 3 days before the experiment. 4. These results suggest that DNP can act within the central nervous system to increase renal water excretion. PMID- 11906483 TI - Effect of vasodilatory beta-adrenoceptor blockers on cardiovascular haemodynamics in anaesthetized rats. AB - 1. The effect of vasodilatory beta-adrenoceptor blockers on regional blood flow in the major organs of anaesthetized rats was investigated using radioactive microspheres. The administration of propranolol and saline was used as a control. 2. Intravenous injections of carvedilol (2 mg/rat), celiprolol (20 mg/rat) and bopindolol (1 mg/rat) equally decreased systemic blood pressure (SBP) by approximately 20 mmHg, whereas propranolol (1 mg/rat) decreased SBP only slightly but not significantly. 3. Heart rate was significantly decreased by carvedilol, celiprolol, bopindolol and propranolol. 4. Coronary blood flow was markedly increased by carvedilol, but not by the other three drugs. 5. Cardiac output tended to decrease following the administration of all four drugs. 6. Total peripheral vascular resistance was not significantly affected by carvedilol, celiprolol and bopindolol, but was markedly increased following propranolol. 7. Renal blood flow was markedly increased by carvedilol. 8. Blood flow in the brown fat was markedly increased by carvedilol and bopindolol, but not by celiprolol and propranolol. 9. These findings indicate that the newer vasodilatory beta blockers, such as carvedilol and bopindolol, have a beneficial effect on the regional circulation in contrast with the classical beta-blocker propranolol. 10. The regional haemodynamic effects observed in the present study following intravenous injection of the beta-blockers may help explain the clinical experience that vasodilatory beta-blockers increase insulin sensitivity and decrease mortality in patients with congestive heart failure. PMID- 11906484 TI - Val-Tyr as a natural antihypertensive dipeptide can be absorbed into the human circulatory blood system. AB - 1. Intact absorption of the bioactive dipeptide Val-Tyr (VY), with in vivo antihypertensive ability in normotensive human subjects, was investigated. 2. As a result of a single oral administration of VY, the VY absorption curve occurred maximally over the second hour postprandially; a greater than 10-fold higher increment of VY following a dose of 12 mg was observed in the plasma at 2 h compared with the baseline concentration of VY at 0 h (1934 +/- 145 vs 159 +/- 11 fmol/mL plasma, respectively). 3. Plasma VY levels increased with dose administered (3, 6 and 12 mg), suggesting that exogenous VY could be absorbed intact into the human blood depending on the dose. The elimination half time (t1/2) of VY was estimated to be 3.1 h. The area under the curve for the 12 mg VY dose was 9185 +/- 688 fmol small middle doth/mL plasma. PMID- 11906485 TI - Exercise and skeletal muscle gene expression. AB - 1. Skeletal muscle is a complex and heterogenous tissue capable of remarkable adaptation in response to exercise training. The role of gene transcription, as an initial target to control protein synthesis, is poorly understood. 2. Mature myofibres contain several hundred nuclei, all of which maintain transcriptional competency, although the localized responsiveness of nuclei is not well known. Myofibres are capable of hypertrophy. These processes require the activation and myogenic differentiation of mononuclear satellite cells that fuse with the enlarging or repairing myofibre. 3. A single bout of exercise in human subjects is capable of activating the expression of many diverse groups of genes. 4. The impact of repeated exercise bouts, typical of exercise training, on gene expression has yet to receive systematic investigation. 5. The molecular programme elicited by resistance exercise and endurance exercise differs markedly. Muscular hypertrophy following resistance exercise is dependent on the activation of satellite cells and their subsequent myogenic maturation. Endurance exercise requires the simultaneous activation of mitochondrial and nuclear genes to enable mitochondrial biogenesis. 6. Future analysis of the regulation of genes by exercise may combine high-throughput technologies, such as gene-chips, enabling the rapid detection and analysis of changes in the expression of many thousands of genes. PMID- 11906486 TI - Large artery stiffness: implications for exercise capacity and cardiovascular risk. AB - 1. Large artery stiffness, or its inverse, compliance, determines pulse pressure, which, in turn, influences myocardial work capacity and coronary perfusion, both of which impact on exercise capacity and cardiovascular risk. 2. In support of a role for arterial properties in exercise performance, aerobically trained athletes (aged 30-59 years) have lower arterial stiffness than their sedentary counterparts. Furthermore, in healthy older subjects (aged 57-80 years), time to exhaustion on treadmill testing correlated positively with arterial compliance. 3. Arterial stiffness is more closely linked to exercise capacity and myocardial risk in patients with coronary disease where, independently of degree of coronary disease, those with stiffer proximal arteries have a lower exercise-induced ischaemic threshold. 4. Moderate aerobic training elevates resting arterial compliance by approximately 30%, independently of mean pressure reduction, in young healthy individuals but not in isolated systolic hypertensive patients. Rat training studies support a role for exercise training in structural remodelling of the large arteries. 5. High-resistance strength training is associated with stiffer large arteries and higher pulse pressure than matched controls. 6. Large artery stiffness is an important modulator of the myocardial blood supply and demand equation, with significant ramifications for athletic performance and ischaemic threshold in coronary disease patients. Moderate aerobic training, but not high-resistance strength training, reduces large artery stiffness in young individuals whereas older subjects with established isolated systolic hypertension are resistant to such adaptation. PMID- 11906487 TI - Adaptations of skeletal muscle to prolonged, intense endurance training. AB - 1. Endurance exercise induces a variety of metabolic and morphological responses/adaptations in skeletal muscle that function to minimize cellular disturbances during subsequent training sessions. 2. Chronic adaptations in skeletal muscle are likely to be the result of the cumulative effect of repeated bouts of exercise, with the initial signalling responses leading to such adaptations occurring after each training session. 3. Recently, activation of the mitogen-activated protein kinase signalling cascade has been proposed as a possible mechanism involved in the regulation of many of the exercise-induced adaptations in skeletal muscle. 4. The protein targets of AMP-activated protein kinase also appear to be involved in both the regulation of acute metabolic responses and chronic adaptations to exercise. 5. Endurance training is associated with an increase in the activities of key enzymes of the mitochondrial electron transport chain and a concomitant increase in mitochondrial protein concentration. These morphological changes, along with increased capillary supply, result in a shift in trained muscle to a greater reliance on fat as a fuel with a concomitant reduction in glycolytic flux and tighter control of acid base status. Taken collectively, these adaptations result in an enhanced performance capacity. PMID- 11906488 TI - Pharmacological profile of nociceptin/orphanin FQ receptors. AB - 1. Nociceptin/orphanin FQ (NC) and its receptor (OP4) represent a novel peptide/receptor system pharmacologically distinct from classical opioid systems. 2. Via OP4 receptor activation, NC regulates several biological functions, both at peripheral and central levels; therefore, the OP4 receptor may be viewed as a novel target for drug development. However, the pharmacology of this receptor is still in its infancy, with few molecules interacting selectively with this receptor. 3. In the present article, we review the findings of studies that have investigated the pharmacological profile of ligands selective for the OP4 receptor, these being two antagonists, the peptide [Nphe1]NC(1-13)NH2 and the non peptide J-113397, and two agonists, the peptide [Arg14,Lys15]NC, and the non- peptide Ro 64-6198. 4. The results of these studies indicate that agents that selectively activate or block the OP4 receptor may represent new potential drugs for the treatment of human diseases. PMID- 11906489 TI - Role of nitric oxide in mediating vasodilator responses to opioid peptides in the rat. AB - 1. Endomorphins 1 and 2, endogenous ligands for the mu-opioid receptor, and nociceptin (orphanin FQ; OFQ), an endogenous ligand for the ORL1 receptor, have vasodilator activity in the vascular bed of the hindquarters of the rat. In the present study, the role of nitric oxide (NO), vasodilator prostaglandins and the opening of KATP channels in mediating vasodilator responses to these novel agonists was investigated in the rat. 2. Under constant-flow conditions, injections of endomorphins 1 and 2, PL017 ([N-MePhe3,D-Pro4]-morphiceptin), nociceptin and Tyr-D-Ala-Gly-MePhe-Gly(ol)-enkephalin (DAMGO) produced dose dependent decreases in hindquarters perfusion pressure. Vasodilator responses to endomorphin 1 and 2, acetylcholine and adrenomedullin, were attenuated by the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) at a time when vasodilator responses to nociceptin, adrenomedullin and the NO donor diethylamine/NO were not altered. 3. Vasodilator responses to endomorphins 1 and 2, nociceptin, PL017 and DAMGO were not altered after administration of sodium meclofenamate at a time when vasodilator responses to arachidonic acid were reduced significantly or after administration of U-37883A at a time when vasodilator responses to levcromakalim were reduced significantly. 4. The results of these studies indicate that vasodilator responses to endomorphins 1 and 2, PL017 and DAMGO are mediated, in large part, by the release of NO, whereas vasodilator responses to nociceptin are mediated by an L-NAME-insensitive mechanism. Moreover, these results demonstrate that the vasodilator responses to these peptides are not due to the release of vasodilator prostaglandins or the opening of KATP channels in the hindquarters vascular bed of the rat. PMID- 11906490 TI - Sympathoinhibitory action of nociceptin in the rat spinal cord. AB - 1. Whole-cell patch recordings were made from antidromically identified sympathetic preganglionic neurons (SPN) of immature rat spinal cord slices. Bath application of nociceptin (0.1-1 micromol/L) suppressed excitatory postsynaptic potentials (EPSP) and hyperpolarized a population of SPN; these effects were naloxone (1 micromol/L) insensitive. 2. Nociceptin suppressed the amplitude of EPSP without causing a concomitant change in glutamate-induced depolarizations, suggesting a presynaptic inhibitory action. 3. Analysis of current-voltage relationships showed that nociceptin hyperpolarized SPN by increasing an inwardly rectifying K+ current. 4. Intrathecal injection of nociceptin (3, 10 and 30 nmol) to urethane-anaesthetized rats dose-dependently reduced the mean arterial pressure and heart rate; these effects were not prevented by prior intravenous injection of naloxone (1 mg/kg). 5. Results from our in vitro and in vivo experiments suggest that nociceptin suppresses spinal sympathetic outflow either by attenuating excitatory synaptic responses or hyperpolarizing SPN. PMID- 11906491 TI - Opioid signalling in the rat rostral ventrolateral medulla. AB - 1. The present article reviews several aspects of opioid signalling in the rostral ventrolateral medulla (RVLM) and their implications for the neural control of blood pressure. 2. In the RVLM, preproenkephalin (PPE) mRNA is expressed by bulbospinal cells that are strongly barosensitive. These putative presympathetic neurons includes C1 and non-C1 neurons. 3. In the RVLM, PPE mRNA is also present in GABAergic neurons that do not project to the thoracic spinal cord. 4. Rostral ventrolateral medulla presympathetic cells receive enkephalinergic inputs and express mu-opioid receptors (MOR). Some of their synaptic inputs also contain MOR. 5. Pre- and post-synaptic modulation of RVLM presympathetic neurons by MOR agonists has been demonstrated in slices of neonate brain. The post-synaptic effect is inhibitory (increased gK). Presynaptic effects include disfacilitation (reduction of glutamate release) and possibly dishinhibition (reduction of GABA release). 6. In conclusion, opioid signalling plays a pervasive role in the medullospinal network that controls sympathetic tone and arterial pressure. Opioid peptides are made by the presympathetic, presumably excitatory, cells of the RVLM and by local GABAergic inhibitory neurons. In addition, RVLM presympathetic neurons are also controlled by opioid peptides at the pre- and post-synaptic level. mu-Opioid receptors are found post synaptically, whereas presynaptic receptors probably include both mu and delta subtypes. Conditions that trigger the release of opioid peptides by presympathetic neurons or by inputs to these cells are not fully understood and may include decompensated haemorrhage and certain types of peripheral sensory stimulation related to acupuncture. PMID- 11906492 TI - Responses to microinjections of endomorphin and nociceptin into the medullary cardiovascular areas. AB - 1. Cardiovascular effects of microinjections of nociceptin and endomorphin-2 into the following medullary areas were studied in urethane-anaesthetized rats: chemoreceptor projection site (CPS), intermediate portion of the nucleus tractus solitarius (I-NTS), caudal ventrolateral medullary depressor area (CVLM) and rostral ventrolateral medullary pressor area (RVLM). 2. Microinjections of nociceptin or endomorphin-2 (0.6 mmol/L each) into the CPS and RVLM elicited depressor and bradycardic responses, whereas similar injections into the I-NTS and CVLM elicited pressor and tachycardic responses. 3. The mechanism of cardiovascular responses to microinjections of these opioid peptides into different medullary areas involved in cardiovascular function can be postulated as follows: the direct effect of nociceptin and endomorphin-2 on neurons is usually inhibitory. Because the activation of CPS and RVLM by microinjections of L-glutamate results in pressor and tachycardic responses, inhibition of these areas by nociceptin and endomorphin-2 elicits depressor and bradycardic responses. Similarly, activation of neurons in the I-NTS and CVLM by microinjections of L-glutamate elicits depressor and bradycardic responses. Therefore, inhibition of these areas by microinjections of these opioid peptides elicits an increase in blood pressure and heart rate. PMID- 11906493 TI - Stress-specific opioid modulation of haemodynamic counter-regulation. AB - 1. The haemodynamic and cardiovascular responses to stress, in addition to being under control of the autonomic nervous system, are also under opiate modulation. Our studies have provided evidence for activation of the endogenous opioid system in haemorrhagic shock, sepsis and trauma. Furthermore, we have demonstrated that both central and systemic opiate administration to naive rats result in marked alterations in haemodynamic responses, which are associated with activation of the sympathetic nervous system. 2. Because of the ubiquitous presence of opiate receptors in both the central nervous system and peripheral tissues, as well as their production and release centrally and peripherally, this facilitates an endocrine as well as a paracrine contribution to modulating vascular responses to stress, either directly or indirectly. Results from previous studies suggest that endogenous opioids are not involved in mediating the lipopolysacharide-induced hypotensive response. 3. In more recent studies, we have examined the role of opiate receptor activation in modulating the haemodynamic and neuroendocrine responses to fixed pressure haemorrhagic shock in conscious unrestrained rats. Using systemic opiate blockade (naltrexone, 15 mg/kg, i.p.) prior to haemorrhage, we have observed that blood loss required to achieve mean arterial blood pressure of 40 mmHg was higher in naltrexone-treated animals than in time-matched saline controls. Interestingly, the haemodynamic modulation exerted by naltrexone cannot be attributed to differences in circulating catecholamine levels. Haemorrhage produced an immediate and progressive increase in circulating adrenaline and noradrenaline levels, reaching values that were 50- and 20-fold higher than basal, respectively. Naltrexone pretreatment did not alter the time-course or magnitude of the rise in circulating levels of catecholamines. 4. These results indicate that endogenous opioid activation contributes to the haemodynamic dishomeostasis associated with blood loss. Our findings suggest stress-specific roles for opiate-sensitive haemodynamic counter-regulatory responses. PMID- 11906495 TI - Chlamydia pneumoniae and atherosclerosis -- what we know and what we don't. AB - The clinical manifestations of atherosclerosis include coronary artery disease (CAD), stroke, abdominal aortic aneurysm and peripheral vascular disease. World wide, CAD and stroke are the leading causes of death and disability. The recognition of atherosclerosis as an inflammatory disease in its genesis, progression and ultimate clinical manifestations has created an interesting area of vascular research. Apart from those well-known traditional risk factors for atherosclerosis, novel and potentially treatable atherosclerotic risk factors such as homocysteine (an amino acid derived from the metabolism of dietary methionine that induces vascular endothelial dysfunction) and infections have emerged. In fact, the century-old 'infectious' hypothesis of atherosclerosis has implicated a number of micro-organisms that may act as contributing inflammatory stimuli. Although cytomegalovirus, Helicobacter pylori and Chlamydia pneumoniae are the three micro-organisms most extensively studied, this review will focus on C. pneumoniae. Collaborative efforts from many disciplines have resulted in the accumulation of evidence from seroepidemiological, pathological, animal model, immunological and antibiotic intervention studies, linking C. pneumoniae with atherosclerosis. Seroepidemiological observations provide circumstantial evidence, which is weak in most prospective studies. Pathological studies have demonstrated the preferential existence of C. pneumoniae in atherosclerotic plaque tissues, while animal model experiments have shown the induction of atherosclerosis by C. pneumoniae. Finally, immunological processes whereby C. pneumoniae could participate in key atherogenic and atherothrombotic events have also been identified. Although benefits of the secondary prevention of atherosclerosis have been demonstrated in some antibiotic intervention studies, a number of negative studies have also emerged. The results of the ongoing large prospective human antibiotic intervention trials may help to finally establish if there is a causal link between C. pneumoniae infection and atherosclerosis. PMID- 11906494 TI - Nociceptin/orphanin FQ modulates the cardiovascular, but not renal, responses to stress in spontaneously hypertensive rats. AB - 1. The central administration of the endogenous opioid-like peptide nociceptin/orphanin FQ (N/OFQ) produces marked cardiovascular depressor and renal sympathoinhibitory responses in conscious animals. These findings are evidence that central N/OFQ may modulate the cardiovascular and renal responses to acute environmental stress. 2. The changes in cardiovascular and renal function produced by intracerebroventricular (i.c.v.) N/OFQ were measured in conscious spontaneously hypertensive rats (SHR) under basal conditions and during the acute environmental stimulus of air jet stress. 3. In SHR, central N/OFQ produced profound hypotensive, bradycardic, renal sympathoinhibitory (delayed) and water diuretic effects by a pathway that does not involve activation of central alpha2 adrenoceptors or classical opioid receptors. 4. Intracerebroventricular injection of N/OFQ prevented the pressor response and blunted the tachycardia to air jet stress. A similar renal sympathoexcitatory and antinatriuretic response was observed in conscious SHR during air stress, before and after i.c.v. N/OFQ. 5. These findings are evidence that, in conscious SHR, i.c.v. N/OFQ selectively inhibited the neural responses to air jet stress by attenuating sympathetic outflow to the heart and, potentially, vasculature, but not to the kidneys. Central endogenous N/OFQ systems may be activated and contribute to regional changes in sympathetic outflow during acute stress. PMID- 11906496 TI - Clinafloxacin monotherapy (CI-960) versus ceftazidime plus amikacin for empirical treatment of febrile neutropenic cancer patients. AB - OBJECTIVE: To assess the efficacy and safety of clinafloxacin as a single agent for the empirical treatment of febrile episodes and bacterial infections in neutropenic cancer patients. METHODS: An open label, active-controlled, randomized, parallel treatment, multicenter study was conducted where clinafloxacin monotherapy was compared to the combination of ceftazidime plus amikacin (plus optional vancomycin or teicoplanin). Four hundred and nineteen patients were randomized to receive either intravenous clinafloxacin 200 mg every 12 h or intravenous ceftazidime (2 g) iv every 8 h plus intravenous amikacin (15 mg/kg) per day in divided doses. All randomized patients were to receive a minimum of 48 h of primary study drug treatment, after which the primary treatment could be modified. Clinical and microbiological responses were evaluated at 7-21 days post-treatment after study treatment and long term (maximum 28 days), in intent-to-treat and modified intent-to-treat populations. RESULTS: Clinafloxacin and ceftazidime-amikacin were statistically equivalent for the 72-h defervescence rate, overall defervescence rate, time to defervescence, clinical success rate, by-pathogen microbiological eradication rate, and survival rate. Clinical cure was achieved in 84% (59/70) of patients who received clinafloxacin monotherapy. There were no significant differences between treatments in rates of adverse events or treatment discontinuation rates due to adverse events. CONCLUSIONS: Clinafloxacin appears to be an appropriate agent for empirical treatment in febrile neutropenic cancer patients. PMID- 11906497 TI - Effect of moxifloxacin on secretion of cytokines by human monocytes stimulated with lipopolysaccharide. AB - OBJECTIVE: To determine the effect of moxifloxacin on secretion of cytokines by human monocytes stimulated with lipopolysaccharide (LPS) or Pansorbin. METHODS: Monocytes obtained from 10 healthy volunteer donors were stimulated with LPS or Pansorbin and exposed or not to different concentrations of the fluoroquinolone antibiotic moxifloxacin. At 3, 6 and 24 h, the amounts of interleukin-1alpha (IL 1alpha), IL-1beta, IL-6, IL-10, IL-12 (p70) and tumour necrosis factor-alpha (TNF alpha) were measured in the supernatants of the monocyte cultures using enzyme linked immunosorbent assay. RESULTS: Stimulation of human monocytes with either LPS or Pansorbin resulted in a significant increase in secretion of each of the cytokines examined. Treatment of LPS-stimulated monocytes with moxifloxacin significantly inhibited (P < 0.01) secretion of IL-1alpha by monocytes of each of 10 human donors; the secretion of TNF-alpha was significantly inhibited (P < 0.01) in monocytes from six of 10 donors. In general there was a trend towards inhibition of secretion of IL-6, IL-10 and IL-12 (p70), but the inhibitory effect was not statistically significant. Secretion of cytokines by Pansorbin-stimulated monocytes was not significantly inhibited by moxifloxacin. CONCLUSIONS: Moxifloxacin has immunomodulatory activity through its capacity to alter the secretion of IL-1alpha and TNF-alpha by human monocytes. PMID- 11906498 TI - Influence of gender and age on course of infection and cytokine responses in mice with disseminated Cryptococcus neoformans infection. AB - OBJECTIVE: To study the influence of gender and age on the course of infection and the cytokine response in a murine model of disseminated cryptococcosis. METHODS: The course of the infection (survival and fungal load in blood and tissues) as well as pro-inflammatory and anti-inflammatory cytokine responses in plasma and organs were compared according to gender and age in outbred mice previously infected with Cryptococcus neoformans NIH52D. RESULTS: Although survival and fungal load were similar in male and female mice, the expression of all cytokines in plasma and of tumour necrosis factor-alpha and interferon-gamma in spleen was significantly increased in female mice compared to male mice in two independent experiments. Young male mice had a significantly shortened survival, were significantly more infected and had predominant tumour necrosis factor-alpha and interferon-gamma responses in comparison with older male mice. CONCLUSION: Host factors should be taken into account when studying the immune response to experimental C. neoformans infection. Our data support epidemiological and clinical data showing differences in susceptibility to cryptococcosis according to gender and age. PMID- 11906499 TI - Direct identification and susceptibility testing of enteric bacilli from positive blood cultures using VITEK (GNI+/GNS-GA). AB - OBJECTIVE: To study the possibility of reporting results of identification and susceptibility testing of Gram-negative bacilli the same day as bacteremia is detected by using direct inoculation from positive blood cultures (Bactec 9240) into VITEK GNI+ and GNS-GA cards. METHODS: All blood cultures with Gram-negative enteric bacillus-like morphology on microscopy found to be positive on workdays between 15 June 1999 and 29 February 2000 were included. Identification and susceptibility testing were done by three methods: the direct method using a suspension made by differential centrifugation of positive blood culture broth for inoculation of the VITEK cards; the standard method using an inoculum made from an overnight culture on a solid media; and the routine method (reference method) using conventional testing. RESULTS: Of 169 isolates, the direct method resulted in 75% correct identifications, 9% misidentifications and 17% non identifications. All misidentified isolates were Escherichia coli, of which 80% were reported as Salmonella arizonae. Five biochemical tests yielded most of the aberrant results; correcting the citrate and malonate reactions in most cases led to correct identification by the VITEK database. Despite a negative H2S reaction, 11 E. coli isolates were reported as S. arizonae. Two-thirds (69%) of identifications were reported within 6 h, and 95% of these were correct. The direct susceptibility testing method was assessable for 140 isolates. Correct results were found in 99% of isolate-antimicrobial combinations, and 85% were reported within 6 h. CONCLUSION: The direct VITEK method could correctly report identifications and susceptibility patterns within 6 h, making same-day reporting possible for almost two-thirds (63%) of bacteremic episodes with Gram-negative bacilli. These results could probably be improved by modification of the identification algorithms of the VITEK software. PMID- 11906500 TI - Clinical microbiological case: facial inflammation in a child with a subnormal serum concentration of IgG2. PMID- 11906501 TI - Differential invasion of respiratory epithelial cells by members of the Burkholderia cepacia complex. AB - To investigate whether there are differences between members of the Burkholderia cepacia complex in their ability to invade human respiratory epithelial cells, 11 strains belonging to genomovars I-V were studied in an antibiotic protection assay using the A549 cell line. Strains belonging to genomovars II and III were more invasive than those of genomovars I, IV and V. There was also intra genomovar variation in invasiveness. No correlation between invasiveness and other putative virulence factors of importance in B. cepacia infection in individuals with cystic fibrosis, cable pilus and B. cepacia epidemic strain marker was identified. PMID- 11906502 TI - Single-dose ciprofloxacin versus 3 days of norfloxacin in uncomplicated urinary tract infections in women. AB - A double-blind, randomized controlled study to evaluate the efficacy and safety of a single dose of 500 mg of ciprofloxacin versus 3 days 400 mg twice-daily of norfloxacin in uncomplicated urinary tract infection in women was designed. Patients underwent four visits: baseline, and at 3, 7 and 28 days. The main efficacy variables were clinical and microbiological outcome at day 7. Analyses on both valid patients and intention-to-treat populations were performed. Two hundred and twenty-six patients (114 receiving ciprofloxacin and 112 receiving norfloxacin) were considered valid for efficacy evaluation. Bacteriologic cure was 91.2% in the ciprofloxacin group and 91.9% in the norfloxacin group. Clinical resolution was 91.2 and 93.8%, respectively. Both treatments were equally efficacious (P = 0.016). PMID- 11906503 TI - Pasteurella gallinarum neonatal meningitis. AB - A 4-day-old baby weighing 1.7 kg was admitted to the neonatal intensive care unit of Ga-Rankuwa Hospital, Pretoria, with a history of apneic attacks. On examination there was an umbilical sepsis and the neonate was septicemic. The baby had been delivered at home and the umbilical cord had been cut by the grandmother using unclean scissors and chimney soot applied to the umbilical stump. On admission, a septic screen was done and antibiotic treatment was started with penicillin and amikacin. The investigations showed that the baby was slightly anemic, with hemoglobin levels of 10.0 g/dL (14.9-23.7 g/dL), and a pure growth of a Gram-negative bacillus was obtained from the cerebrospinal fluid, blood culture and suprapubic aspirate urine specimens. The Gram-negative bacillus was catalase and oxidase positive and it was identified as Pasteurella gallinarum. Antimicrobial profiling showed the organism to be susceptible to penicillin, cefotaxime, gentamicin and amikacin. Despite having received antimicrobial agents to which the etiological agent was susceptible, the neonate died within 5 days of admission. The cause of death was postulated to be due to overwhelming sepsis which resulted in septic shock. PMID- 11906505 TI - Neural networks in human epilepsy: evidence of and implications for treatment. PMID- 11906504 TI - Increased expression of the neuronal glutamate transporter (EAAT3/EAAC1) in hippocampal and neocortical epilepsy. AB - PURPOSE: To define the changes in gene and protein expression of the neuronal glutamate transporter (EAAT3/EAAC1) in a rat model of temporal lobe epilepsy as well as in human hippocampal and neocortical epilepsy. METHODS: The expression of EAAT3/EAAC1 mRNA was measured by reverse Northern blotting in single dissociated hippocampal dentate granule cells from rats with pilocarpine-induced temporal lobe epilepsy (TLE) and age-matched controls, in dentate granule cells from hippocampal surgical specimens from patients with TLE, and in dysplastic neurons microdissected from human focal cortical dysplasia specimens. Immunolabeling of rat and human hippocampi and cortical dysplasia tissue with EAAT3/EAAC1 antibodies served to corroborate the mRNA expression analysis. RESULTS: The expression of EAAT3/EAAC1 mRNA was increased by nearly threefold in dentate granule cells from rats with spontaneous seizures compared with dentate granule cells from control rats. EAAT3/EAAC1 mRNA levels also were high in human dentate granule cells from patients with TLE and were significantly elevated in dysplastic neurons in cortical dysplasia compared with non-dysplastic neurons from postmortem control tissue. No difference in expression of another glutamate transporter, EAAT2/GLT-1, was observed. Immunolabeling demonstrated that EAAT3/EAAC1 protein expression was enhanced in dentate granule cells from both rats and humans with TLE as well as in dysplastic neurons from human cortical dysplasia tissue. CONCLUSIONS: Elevations of EAAT3/EAAC1 mRNA and protein levels are present in neurons from hippocampus and neocortex in both rats and humans with epilepsy. Upregulation of EAAT3/EAAC1 in hippocampal and neocortical epilepsy may be an important modulator of extracellular glutamate concentrations and may occur as a response to recurrent seizures in these cell types. PMID- 11906506 TI - Familial temporal lobe epilepsy with aphasic seizures and linkage to chromosome 10q22-q24. AB - PURPOSE: To describe the phenotypic expression of a new family with familial lateral temporal lobe epilepsy with aphasic seizures, and to compare the findings with the clinical features of previously reported families linked to chromosome 10q22-q24. METHODS: Medical records were collected from 12 living affected members. The patients underwent a personal interview and a clinical neurologic examination. Results from interictal scalp EEGs and neuroimaging examinations were obtained. RESULTS: The cardinal ictal symptom was a brief sensory aphasia in eight of the patients. In four, this was accompanied by auditory symptoms, usually in the form of monotonous unformed sounds. Simple partial seizures with psychic or somatosensory seizures also were present. Visual ictal symptoms and complex partial seizures were absent. All patients had generalized tonic-clonic seizures. Magnetic resonance imaging (MRI) or computed tomography (CT) did not reveal morphologic correlates. Improvement with age seemed to occur in many patients. Significant linkage to chromosome 10q22-q24 was established by testing 17 polymorphic microsatellite markers. CONCLUSIONS: The epilepsy of this family appears to represent a variety of autosomal dominant lateral temporal lobe epilepsy. Aphasic seizures and a peculiar seizure-precipitating effect of the activation of speech (initiation or perception) may serve as markers for identifying further families with this phenotype. PMID- 11906507 TI - An association between mesial temporal lobe epilepsy with hippocampal sclerosis and human leukocyte antigens. AB - PURPOSE: Mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS) is one of the medically intractable epilepsies that may be remediable with surgery. Although the pathogenesis of HS still remains obscure, genetics may play a role as a predisposing factor, with the genetically controlled immune system as one of its aspects. Our aim in this study was to investigate whether there is any association between human leukocyte antigens (HLAs) that are related to chromosome 6 and this specific type of epilepsy. METHODS: HLA class I and II typing were performed with the microlymphocytotoxicity method on 65 Turkish patients with MTLE-HS and on 184 healthy controls. RESULTS: Our study revealed a significantly high frequency of class II antigens HLA-DQ2, -DR4, and -DR7 alleles and the combination of HLA-DR4-DQ2, and DR7-DQ2 alleles. CONCLUSIONS: The HLA alleles that occur with increased frequency in many HLA- associated conditions appear to serve as risk factors that increase susceptibility but are not essential for disease expression. Our data support the role of genetic factors in the development of HS, possibly related to early childhood events that may act as a trigger factor to initiate the cascade in genetically prone patients with specific HLA types to give rise to MTLE eventually. PMID- 11906508 TI - Neocortical seizure termination by focal cooling: temperature dependence and automated seizure detection. AB - PURPOSE: The therapy for focal neocortical epilepsy remains suboptimal. We have, therefore, worked to develop techniques to cool small regions of the neocortical surface for seizure mapping and, ultimately, for long-term suppression of focal seizures. METHODS: We induced focal neocortical seizures in halothane anesthetized rats by the microinjection of 4-aminopyridine (4-AP) into the motor cortex. The dura over the injection site was cooled with a Peltier device, and the temperature at the interface between dura and Peltier was measured with a thermocouple. In some experiments, seizures were automatically detected by a computer program that activated the Peltier device. RESULTS: Monopolar EEG indicated that our seizures were focal and suppressed when cooling was applied directly over the injection site. The threshold temperature required to observe any reduction in seizure duration was 24 degrees C. The temperature gradient across the cooled neocortex was sharp, with the temperature increasing to 31 degrees C at 4 mm below the Peltier, which was cooled to 20 degrees C. Automatic seizure detection reduced the total seizure duration from 43.4 +/- 33.6 s to 5.6 +/- 5.3 s. CONCLUSIONS: Cooling terminates neocortical seizures when applied very close to the epileptogenic focus. The threshold for seizure termination (24 degrees C) may be lower than the threshold for termination of normal cortical activity, suggesting that this technique will not dissociate the anticonvulsant effect of cooling from the disruption of normal behavior. However, when coupled with automatic seizure detection, focal cooling remains an attractive option for development as a treatment for focal epilepsy. PMID- 11906509 TI - Informative value of magnetic resonance imaging and EEG in the prognosis of infantile spasms. AB - PURPOSE: To investigate the informative value of EEG and cranial magnetic resonance imaging (cMRI) in the prognosis of infantile spasms (ISs); 86 patients with ISs were included in this study. METHODS: All cases had epileptic spasms, psychomotor retardation, and hypsarrhythmia in at least one of their EEGs. cMRIs and laboratory tests necessary for etiologic diagnosis were completed in all cases. Patients were followed up periodically both clinically and by video-EEGs for >1 year. Clinical information was categorized on the basis of four spheres as epilepsy, psychosocial development, motor development, and overall clinical condition, with each category being evaluated under three levels of involvement as good, moderate, and severe, depending on selected parameters. A similar scale was applied for the EEG results and for the cMRI findings. Clinical parameters were correlated to EEG and cMRI results, by Spearman test. Other statistical tests used were Kruskal-Wallis chi(2) and Mann-Whitney U analysis as multiple comparison by post hoc Bonferroni correction. RESULTS: A severe overall clinical course was observed in 64% of patients, whereas this incidence was 58% and 44% in the EEG follow-up and cMRI parameters, respectively. In regard to prognosis, a significant correlation was determined between the clinical and the EEG course. This relation was the most prominent in psychosocial developmental parameters and least prominent in the motor development. cMRI findings, however, were correlated only with motor development. CONCLUSIONS: cMRI and repeated EEG recordings, especially when assessed together, may provide complementary information regarding the prognosis in ISs. PMID- 11906510 TI - Interictal and ictal EEG activity in the basal ganglia: an SEEG study in patients with temporal lobe epilepsy. AB - PURPOSE: The interictal and ictal EEG activity in the basal ganglia in patients with temporal lobe epilepsy were studied during invasive EEG monitoring. METHODS: Eight epilepsy surgery candidates, each with a proven mesiotemporal seizure-onset zone, participated in the study. We used two invasive EEG methods to determine the seizure-onset zone. In both methods, diagonal electrodes were targeted into the amygdalohippocampal complex via a frontal approach and were passed through the basal ganglia with several leads. We analyzed 16 partial epileptic seizures, four of which became secondarily generalized. RESULTS: No epileptic interictal or ictal discharges were noticed in the basal ganglia. The interictal activity in the basal ganglia was a mixture of low-voltage beta activity and medium-voltage alpha-theta activity. When the ictal paroxysmal activity remained localized to the seizure-onset zone, the activity of the basal ganglia did not change. The spread of epileptic activity to other cortical structures was associated with the basal ganglia EEG slowing to a theta-delta range of 3-7 Hz. This slowing was dependent on the spread of ictal discharge within the ipsilateral temporal lobe (related to the investigated basal ganglia structures); alternatively, the slowing occurred in association with the regional spread of ictal activity from the mesiotemporal region to the temporal neocortex contralaterally to the investigated basal ganglia. Secondary generalization was associated with a further slowing of basal ganglia activity. CONCLUSIONS: The basal ganglia do not generate specific epileptic EEG activity. Despite the absence of spikes, the basal ganglia participate in changing or reflect changes in the distribution of the ictal epileptic activity. PMID- 11906511 TI - Superiority of HMPAO ictal SPECT to ECD ictal SPECT in localizing the epileptogenic zone. AB - PURPOSE: We examined diagnostic performances of Tc-99m hexamethylpropylene amine oxime (HMPAO) and Tc-99m electron capture detection (ECD) ictal single-photon emission computed tomography (SPECT) to localize the epileptogenic zones in mesial temporal lobe epilepsy (TLE) and neocortical epilepsy (NE). METHODS: Epileptogenic zones were identified by invasive EEG or surgical outcome. Ictal SPECT was performed with stabilized Tc-99m HMPAO (TLE, 17; NE, 23) and with Tc 99m ECD (TLE, 7; NE, 7). Single-blind visual interpretation was used to localize the epileptogenic zones. Asymmetric index was calculated. Subtraction ictal SPECT was coregistered to a magnetic resonance imaging (MRI) template. RESULTS: In TLE, the sensitivity of Tc-99m HMPAO SPECT was 82% (14 of 17) and that of Tc-99m ECD SPECT was 71% (five of seven). The asymmetric index (AI; 25 +/- 10) of Tc-99m HMPAO SPECT was larger (p = 0.05) than the AI (13 +/- 13) of Tc-99m ECD SPECT in patients with TLE. In NE, the sensitivity of Tc-99m HMPAO SPECT was 70% (16 of 23), but that of Tc-99m ECD SPECT was 29% (two of seven). The AI (15 +/- 10) of Tc-99m HMPAO SPECT was significantly larger (p = 0.02) than the AI (4.8 +/- 6) of Tc-99m ECD SPECT in patients with NE. Subtraction ictal SPECT coregistered to MRI supported the visual assessment. CONCLUSIONS: We concluded that the sensitivity of Tc-99m ECD ictal SPECT is similar to that of Tc-99m HMPAO ictal SPECT in TLE; however, ictal hyperperfusion was higher with the Tc-99m HMPAO SPECT. In patients with NE, Tc-99m HMPAO ictal SPECT also was superior to Tc-99m ECD ictal SPECT in sensitivity and degree of hyperperfusion. PMID- 11906512 TI - Contribution of SISCOM imaging in the presurgical evaluation of temporal lobe epilepsy related to dysembryoplastic neuroepithelial tumors. AB - PURPOSE: Dysembryoplastic neuroepithelial tumors (DNTs) are a group of glioneuronal supratentorial and intracortical lesions often associated with the early onset of intractable and crippling partial seizures. They are characterized by their location, multinodular architecture, and heterogeneous cell composition, with a specific glioneuronal element in the specific form. Foci of cortical dysplasia may be associated with the tumoral lesion, and identifying the presence and the extent of cortical dysplasia is not always easy on magnetic resonance images (MRIs). The purpose of this article is to evaluate, retrospectively, the usefulness of ictal single-photon emission computed tomography (SPECT) imaging to assess the presence and the extent of cortical dysplasia associated with DNTs in nine patients with intractable temporal lobe epilepsy related to histopathologically confirmed DNTs. METHODS: The results of the subtraction of ictal and interictal SPECT coregistered to MRI (SISCOM) were compared with the results of the examinations of pathological material after surgery. RESULTS: SISCOM showed a strongly hyperperfused area corresponding anatomically to electroclinical abnormalities and to the location of DNTs on MRI. A circumscribed hyperperfusion was present in DNTs without cortical dysplasia, limited to the location of the tumor on MRI. In cases of associated cortical dysplasia, a widespread hyperperfusion including areas corresponding to normal perilesional regions on MRI was found. CONCLUSIONS: SISCOM, used among presurgical investigations, contributes to detecting cortical dysplasia associated with DNTs. Concordance between the symptomatogenic zone (defined from the medical history and electroclinical data), MRI scans, SISCOM pattern, and complete resection of the epileptic zone was predictive of a good postsurgical outcome. PMID- 11906513 TI - Focal ictal beta discharge on scalp EEG predicts excellent outcome of frontal lobe epilepsy surgery. AB - PURPOSE: To determine whether a focal beta-frequency discharge at seizure onset on scalp EEG predicts outcome of frontal lobe epilepsy (FLE) surgery. METHODS: We identified 54 consecutive patients with intractable FLE who underwent epilepsy surgery between December 1987 and December 1996. A blind review of EEGs and magnetic resonance images (MRIs) was performed. Lesional epilepsy is defined as presence of an underlying structural abnormality on MRI. RESULTS: Overall, 28 (52%) patients were seizure free, with a mean follow-up of 46.5 months. Presence of a focal beta-frequency discharge at seizure onset on scalp EEG predicted seizure-free outcome in lesional (p = 0.02) and non-lesional (p = 0.01) epilepsy patients. At least 90% of patients who had either lesional or non-lesional epilepsy were seizure free if scalp EEG revealed a focal beta discharge at ictal onset. Moreover, logistic regression analysis showed that focal ictal beta pattern and completeness of lesion resection were independently predictive of seizure-free outcome. Ictal onset with lateralized EEG activity of any kind and postresection electrocorticographic spikes did not predict surgical outcome (p > 0.05). CONCLUSIONS: Only about 25% of FLE surgical patients have a focal beta frequency discharge at seizure onset on scalp EEG. However, its presence is highly predictive of excellent postsurgical seizure control in either lesional or non-lesional FLE surgical patients. PMID- 11906514 TI - Memory deficits after resection from left or right anterior temporal lobe in humans: a meta-analytic review. AB - PURPOSE: Memory deficits in epileptic patients have been found in some, but not all studies assessing the effects of side of seizures and resection from a temporal lobe on cognitive performance. The purpose of this study was to provide a quantitative review of previous studies on this issue. METHODS: Based on conventional meta-analytic procedures, we identified 33 studies that assessed verbal and nonverbal memory performance before and after anterior temporal lobectomy. The Logical Memory and Visual Reproduction subtests from the Wechsler Memory Scale were used. These studies were then subjected to two levels of analyses: (a) vote-counting procedure, and (b) effect-size calculations and comparisons. RESULTS: Overall, the data confirmed previous findings that verbal memory tasks are sensitive to left hemisphere dysfunction. The efficacy of a "nonverbal" task for tapping function in the nondominant (right) hemisphere was not confirmed, although a trend supporting this speculation was observed. With regard to the comparison of changes in verbal and nonverbal memory before and after resection from a temporal lobe, a clear trend was observed for decline in verbal memory function after resection from the left, especially significant for immediate verbal recall. A trend for contralateral improvement on nonverbal memory also was observed. The pattern of memory change after resection from the right temporal lobe was less clear. CONCLUSIONS: The findings of this study suggest that side of epileptic seizure and surgical resection from a temporal lobe affect verbal memory functions. The relations between the laterality of epileptic seizure, surgical resection from the temporal lobe, and nonverbal memory are to be verified by further research. PMID- 11906515 TI - The use of stereotactic radiosurgery to treat intractable childhood partial epilepsy. AB - PURPOSE: Although conventional surgery is presently used to treat seizures of temporolimbic and neocortical origin, deep-seated lesions are often associated with morbidity. Stereotactic radiosurgery is a noninvasive procedure that effectively treats patients with vascular malformations and brain tumors, but its efficacy for epileptogenic foci is limited, especially in children. METHODS: Between 1995 and 1999, four candidates who had medically uncontrolled seizures and localized seizure foci were selected for stereotactic radiosurgery, with a mean age of 9.75 years at the time of surgery (range, 4-17 years). Seizure foci were identified on the basis of ictal and interictal video-EEG. Magnetic resonance (MR) images were obtained before and after surgery. Ictal single-photon emission computed tomography (SPECT) was performed by using stabilized hexamethyl propyleneamine oxime (HMPAO; 300 microcuries/kg) with early injection after electrographic ictal onset. The clinical features of the patients are given. All radiosurgical procedures were performed with the gamma knife unit with the Leksell stereotactic frame, stereotactic MRI imaging, and the Gamma Plan workstation. Seizure outcome was scored according to Engel's classification. RESULTS: Two patients had hypothalamic hamartoma (HH), and two had neocortical epilepsy. At mean follow-up of 39.2 months (range, 26-69 months), two patients were seizure free, one with a HH and one with a suggestive developmental tumor in the insular cortex by MRI findings. The other patient with HH had 90% reduction of seizures. One patient with a widespread seizure focus that involved the motor strip was unimproved. The two patients with HH also exhibited markedly improved neurobehavioral status after surgery. There were no significant complications of radiosurgical therapy. CONCLUSIONS: Our findings suggest that gamma knife surgery is a potentially valuable treatment modality for children with medically intractable epilepsy due to a well-localized seizure focus that is difficult to excise by conventional techniques or for whom they are deemed unsuitable. More widespread application in childhood epilepsy should be investigated in larger series. PMID- 11906516 TI - Attention deficits are not characteristic of schoolchildren with newly diagnosed idiopathic or cryptogenic epilepsy. AB - PURPOSE: To compare problems of attention in schoolchildren with newly diagnosed idiopathic or cryptogenic epilepsy with those in healthy classmates. METHODS: A computerized battery of tasks comprised Reaction Time (RT) measurement, Trail making (Color Trails 1 and 2), Manual Tapping and Steadiness, and a newly developed task of sustained attention (Balloon Piercing). SUBJECTS: Fifty-one children with epilepsy (age 7-16 years) and 48 gender- and age-matched classmates were assessed thrice: within 48 h after diagnosis [before start of antiepileptic drugs (AEDs)], and 3 and 12 months later. Significantly more children with epilepsy (51%) than control children (27%) had required special educational assistance at school. RESULTS: Children with epilepsy could not be distinguished from controls in execution times or motor speed. However, errors were more frequent among patients in a "go-no-go" RT task, and errors of omission in a task requiring sustained attention. Within the group of children with epilepsy, those with prior school or behavior difficulties and those whose parents reacted maladaptively to the onset and diagnosis of epilepsy performed worse than those without these adversities, in the sense that their RT increased inordinately with increasing task difficulty. Epilepsy-related variables did not explain any variance. Transient inordinately poor performances were found in 69% of patients and 40% of controls. CONCLUSIONS: Children with newly diagnosed "epilepsy only" do not have persistent attention deficits. AED treatment has no detrimental effect on attention. Prior school and behavior difficulties and a maladaptive reaction to the onset of epilepsy rather than epilepsy variables are related to decreased attentional efficiency. PMID- 11906517 TI - Public awareness, attitude, and understanding of epilepsy in Hong Kong Special Administrative Region, China. AB - PURPOSE: Because of the nature of the epileptic seizure, the social stigma attached to epilepsy is a major handicap to persons with epilepsy compared with the disability associated with seizures or the side effects from medications. Measuring the awareness, attitude, and understanding of epilepsy is the first step in alleviating discrimination. METHODS: We conducted a face-to-face questionnaire interview survey in five different locations (HKSAR) that represented the population structure, administrative function, and occupations of inhabitants. Subjects with epilepsy or with relatives who had epilepsy were excluded. RESULTS: We interviewed 1,128 subjects; 58.2% had heard about epilepsy before. Of these, 55% had witnessed one or more epileptic seizure, and 18.9% knew one or more persons with epilepsy; 52.7% would put an object into a patient's mouth during an epileptic seizure to prevent injury of the tongue (32.2% learned this from a local television program), and 94.1% agreed that persons with epilepsy could be married. However, only 72.5% considered pregnancy to be appropriate; 11.2% would not let their children play with others with epilepsy; 32.2% would not allow their children to marry persons with epilepsy. Employers (22.5%) would terminate the employment contract after an epileptic seizure in an employee with unreported epilepsy. CONCLUSIONS: This study documented the public attitude toward epilepsy in HKSAR; although it was more negative than that in Western societies, it was more positive than that of the Chinese in China or Taiwan. We suggest that more effort be made to improve public awareness of, attitude toward, and understanding of epilepsy through school education and epilepsy-related organizations in HKSAR. PMID- 11906519 TI - Dystonia, mirror movement, and epileptic seizure. PMID- 11906518 TI - The experience of earthquakes by patients with epileptic and psychogenic nonepileptic seizures. AB - PURPOSE: We sought to understand better the experience of seizures by studying differences in the subjective experience of being in an earthquake between patients with epileptic (EP) and nonepileptic (NES) seizures. METHODS: Forty eight patients with CCTV/EEG-documented EP or NES who were in the Seattle metropolitan area during the February 28, 2001 Nisqually earthquake were randomly selected for telephone interviews on their earthquake experiences, including whether they thought they were having a seizure during the event. RESULTS: Twenty three percent of EP patients spontaneously volunteered that they initially thought they were having a seizure during the earthquake as compared with none of the NES individuals (p = 0.03). However, 35% of EP and 23% of NES patients thought they were having a seizure during it when asked directly (p = 0.37). The most common reasons given, regardless of seizure type, were shaking and feelings of losing control. Of those responding negatively, 100% of EP and 47% of NES patients said that movement of their environment indicated that it was not a seizure (p = 0.001). EP patients took an average of 42 s to realize that the earthquake was not a seizure compared with 105 s for the NES group (p = 0.06). The earthquake precipitated seizures in both groups (11.5% EP, 9.1% NES). CONCLUSIONS: EP patients were more likely to mistake the earthquake spontaneously for a seizure. This indicates these two experiences are similar and provides a glimpse into the subjective experience of a seizure for those who have never had one but have experienced an earthquake. PMID- 11906520 TI - The speeding of EPSC kinetics during maturation of a central synapse. AB - Several factors contribute to the shape of excitatory postsynaptic currents (EPSCs) in CNS neurons, among them the kinetics of presynaptic release, transmitter clearance, and the properties and distribution of postsynaptic receptors. The decays of AMPA receptor-mediated EPSCs at rat cerebellar mossy fibre-granule cell (MF-gc) synapses follow a bi-exponential time-course. The fast component dominates the decay, accounting for 84-94% of the peak amplitude. Here we show that both components of decay, and also the risetimes, became faster during postnatal maturation. At adult, but not immature, synapses, the risetimes and decays of evoked multiquantal EPSCs were similar to those of monoquantal miniature (m)EPSCs. The faster risetimes at mature synapses reflected increased synchrony of multivesicular release, whereas the faster decays appeared to reflect changes in the properties of postsynaptic receptors. Inhibition of glutamate uptake was without effect on evoked EPSCs at both ages. Furthermore, after slowing receptor desensitization with cyclothiazide, the EPSCs at mature synapses decayed as slowly as EPSCs at immature synapses, suggesting that faster glutamate clearance does not account for the developmental speeding of EPSC decay. Our results support previous conclusions that glutamate clearance and receptor deactivation are important determinants of the fast decay component at immature synapses. Desensitization becomes increasingly important during development and plays a major role in shaping EPSC decay at mature synapses. PMID- 11906521 TI - Selective targeting of zebrafish olfactory receptor neurons by the endogenous OMP promoter. AB - The olfactory nervous system of fish, in particular zebrafish, has become a valid model for that of higher vertebrates. However, no genetic markers for olfactory specific cell types, e.g. the olfactory receptor neurons, have been established in this species. Olfactory marker protein (OMP) is a reliable marker for olfactory receptor neurons in several other vertebrates. We have cloned zOMP, the zebrafish homologue of olfactory marker protein. During development, zOMP is expressed exclusively in the olfactory placode, presumably in olfactory receptor neurons, as shown by in situ hybridization. In the adult nasal epithelium zOMP is found restricted to the sensory region. zOMP appears to be a single gene, without close family members. The 5'-flanking region lacks most of the expected regulatory sequence motifs, both general and cell type-specific ones. Nevertheless, it drives reporter gene expression strongly and specifically in olfactory receptor neurons during the whole developmental period examined. Thus the zOMP promoter constitutes a powerful tool which should be useful to selectively introduce a wide variety of genetic modifications into olfactory receptor neurons. PMID- 11906522 TI - Genetic labelling of specific axonal pathways in the mouse central nervous system. AB - We report on transgenic mouse lines in which several sensory systems in the brain are specifically visualized genetically. We employed GAP-lacZ as an axon-targeted reporter protein that was constructed by fusing the membrane-anchoring domain of the GAP-43 protein to lacZ. The reporter gene was introduced into the genome under the control of a promoter element of Brn3b transcription factor to establish transgenic mouse lines. The individual lines thus generated afforded clear images of specific axonal pathways of the visual, vomeronasal, pontocerebellar, and auditory systems. The reporter protein labelled the entire axonal process as well as the cell body of developing and mature neurons on staining with X-gal. We show that these lines facilitate the developmental and anatomical study of these neural systems. This strategy should be applicable to a variety of neural systems by using various specific promoter elements. PMID- 11906523 TI - D2-mediated modulation of N-type calcium currents in rat globus pallidus neurons following dopamine denervation. AB - We have studied the effects of dopamine and the D2-like agonist quinpirole on calcium currents of neurons isolated from the striatum and the globus pallidus (GP). Experiments were performed in young adult rats, either in control conditions or following lesion of the nigrostriatal pathway by the unilateral injection of 6-hydroxydopamine (6-OHDA) in the substantia nigra. Apomorphine driven contralateral turning, 15 days after lesioning, assessed the severity of the dopamine denervation. In addition, the loss of tyrosine hydroxylase immunohistochemistry confirmed the extent of the toxin-induced damage. In both striatal medium spiny (MS) and GP neurons of control animals dopamine and quinpirole promoted a very modest inhibition of calcium conductance. Following 6 OHDA, the inhibition was unaltered in MS (from 10 to 12%), but significantly augmented in GP neurons (21% vs. 9%). Interestingly, analogous inhibition was observed in GP neurons dissociated 20 h after reserpine treatment. Further features of the D2 response were thus studied only in neurons isolated from 6 OHDA-lesioned GP. The D2 modulation was G-protein-mediated but not strictly voltage-dependent. omega-Conotoxin-GVIA occluded the response implying the involvement of N-type calcium channels. The effect of quinpirole developed fast and was insensitive to alterations of cytosolic cAMP. The incubation in phorbol esters or OAG blocked the D2 effect, supporting the involvement of PKC. These findings suggest that postsynaptic D2-like receptors are functionally expressed on GP cell bodies and may supersensitize following dopamine-denervation. A direct D2 modulation of calcium conductance in GP may alter GP firing properties and GABA release onto pallidofugal targets. PMID- 11906524 TI - Periwound dopaminergic sprouting is dependent on numbers of wound macrophages. AB - Injury to many regions of the central nervous system, including the striatum, results in a periwound or 'abortive' sprouting response. In order to directly evaluate whether macrophages play an important role in stimulating periwound sprouting, osteopetrotic (op/op) mice, which when young are deficient in a variety of macrophage subtypes, were given striatal wounds and the degree of dopaminergic sprouting subsequently assessed. Two weeks postinjury, significantly fewer wound macrophages were present in the striata of op/op mice compared with controls (144 +/- 30.1 in op/op mice vs. 416.6 +/- 82.3 in controls, P < 0.005, analysis performed on a section transecting the middle of the wound). Dopamine transporter immunohistochemistry revealed a marked decrease in the intensity of periwound sprouting in the op/op group of animals. Quantification of this effect using [H3]-mazindol autoradiography confirmed that periwound sprouting was reduced significantly in the op/op mice compared with controls (71.4 +/- 21.7 fmol/mg protein in op/op mice vs. 210.7 +/- 27.1 fmol/mg protein in controls, P < 0.0005). In the two groups of animals the magnitude of the sprouting response in individuals was closely correlated with the number of wound macrophages (R = 0.83, R2 = 0.69). Our findings provide strong support for the crucial involvement of macrophages in inducing dopaminergic sprouting after striatal injury. PMID- 11906525 TI - Circadian tryptophan hydroxylase levels and serotonin release in the suprachiasmatic nucleus of the rat. AB - Serotonin (5-HT) plays an important role in the regulation of the time-keeping system in rodents. In the present study, we have investigated the interplay between the rhythms of 5-HT synthesis and release in the suprachiasmatic nuclei (SCN) of the rat. The quantitative distribution of tryptophan hydroxylase (TpH) protein was used as an index of 5-HT synthesis, in perikarya and terminals areas. In the raphe medianus, the maximal levels of TpH was reached in the early daytime period, followed by a decrease before the onset of darkness. Conversely, in the axon terminals of the SCN the highest levels of TpH were found before the onset of the dark-period. Furthermore, TpH amount in SCN displays variations depending on the anatomical area of the SCN. Extracellular 5-HT peaked at the beginning of the night, as evidenced by in vivo microdialysis in the SCN. The 5-HT metabolite, 5-HIAA, presented a similar pattern, but the acrophase occurred in the middle of the dark period. These results suggest that TpH is transported from the soma to the nerve terminals in which 5-HT is synthesized during daytime. This would fill the intracellular stores of 5-HT to provide for its nocturnal release. PMID- 11906526 TI - Altered regulation of the 5-HT system in the brain of MAO-A knock-out mice. AB - Genetic deficiency of monoamine oxidase-A (MAO-A) induces major alterations of mood and behaviour in human. Because serotonin (5-HT) is involved in mood regulation, and MAO-A is responsible for the catabolism of 5-HT, we investigated 5-HT mechanisms in knock-out mice (2-month-old) lacking MAO-A, using microdialysis, electrophysiological, autoradiographic and molecular biology approaches. Compared to paired wild-type mice, basal extracellular 5-HT levels were increased in ventral hippocampus (+202%), frontal cortex (+96%) and dorsal raphe nucleus (DRN, +147%) of MAO-A mutant mice. Conversely, spontaneous firing rate of 5-HT neurons in the DRN (recorded under chloral hydrate anaesthesia) was approximately 40% lower in mutants. Acute 5-HT reuptake blockade by citalopram (0.2 and 0.8 mg/kg i.v.) produced a much larger increase in extracellular 5-HT levels (by approximately 4 fold) and decrease in DRN neuronal firing (with a approximately 4.5 fold decrease in the drug's ED50) in MAO-A knock-out mice, which expressed lower levels of the 5-HT transporter throughout the brain (-13 to -34% compared to wild-type levels). The potency of the 5-HT1A agonist 8-OH-DPAT to produce hypothermia and to reduce the firing of DRN serotoninergic neurons was significantly less in the mutants, indicating a desensitization of 5-HT1A autoreceptors. This was associated with a decreased autoradiographic labelling of these receptors (-27%) in the DRN. Altogether, these data indicate that, in MAO-A knock-out mice, the enhancement of extracellular 5-HT levels induces a down regulation of the 5-HT transporter, and a desensitization of 5-HT1A autoreceptors which allows the maintenance of tonic activity of 5-HT neurons in the DRN. PMID- 11906527 TI - Reduction of drug-induced yawning and penile erection and of noncontact erections in male rats by the activation of GABAA receptors in the paraventricular nucleus: involvement of nitric oxide. AB - The effect of muscimol, a GABAA receptor agonist, injected into the paraventricular nucleus (PVN) of the hypothalamus on drug-induced (apomorphine, oxytocin and NMDA) yawning and penile erection, and on the increase in the concentration of NO2- and NO3- occurring in the paraventricular dialysate in these experimental conditions, was studied in male rats. Muscimol (50, 100 and 200 ng) reduced, in a dose-dependent manner, penile erection and yawning induced by apomorphine (50 ng), oxytocin (30 ng) and NMDA (50 ng) delivered into the PVN. The reduction of penile erection and yawning was parallel to a reduction of the concomitant NO2- and NO3- increase that occurs in the paraventricular dialysate in this experimental condition. In contrast, baclofen (200 ng), a GABAB receptor agonist, was ineffective. The muscimol effects on drug-induced penile erection, yawning and NO2- increase were prevented by the prior administration of bicuculline (250 ng into the paraventricular nucleus). Muscimol (200 ng) but not baclofen (200 ng), injected into the PVN, reduced both noncontact erections in male rats placed in the presence of an inaccessible receptive female, and also the NO2- increase that occurs in the paraventricular dialysate in this experimental condition. As found with drug-induced penile erection, the muscimol reduction of noncontact erections and of NO2- increase was prevented by bicuculline. The present results show that the activation of GABAA receptors in the PVN reduces yawning and penile erection induced by drugs or physiological stimuli by reducing the increase in NO activity that occurs in this hypothalamic nucleus in these experimental conditions. PMID- 11906528 TI - mu/delta Cooperativity and opposing kappa-opioid effects in nucleus accumbens mediated antinociception in the rat. AB - We previously demonstrated that noxious peripheral stimulation (e.g. subdermal capsaicin injection in the hind paw) produces antinociception that is mediated by opioid receptors in nucleus accumbens. The current study used the trigeminal jaw opening nociceptive reflex responses in the rat to assess the role of intra accumbens mu-, delta- and kappa-opioid receptors in the antinociceptive effect of noxious stimulation and intra-accumbens opioid agonism. Whilst intra-accumbens injection of either the mu-receptor-selective antagonist Cys2,Tyr3,Orn5,Pen7amide (CTOP) or the delta-receptor-selective antagonist naltrindole blocked capsaicin induced antinociception, neither the selective mu-agonist [D-Ala2,N-Me-Phe4,Gly5 ol]-enkephalin (DAMGO; 150 or 300 ng) nor the selective delta-agonist D-Pen2,5 enkephalin (DPDPE; 150 or 300 ng) alone induced antinociception. Simultaneous injection of DAMGO and DPDPE (150 ng each), however, produced significant antinociception. Capsaicin-induced antinociception was not blocked by the selective kappa-receptor antagonist nor-binaltorphimine, but was blocked by the kappa-agonist U69,593. U69,593 also antagonized the antinociceptive effect of the DAMGO/DPDPE combination. Thus, in nucleus accumbens, mu- and delta- but not kappa opioid receptors contributed to capsaicin-induced antinociception; selective activation of individual receptor subtypes was insufficient, but coactivation of mu- and delta-opioid receptors induced antinociception, and kappa-receptors appeared to play an antianalgesic role in nucleus accumbens. PMID- 11906530 TI - Orientation specificity of contrast adaptation in visual cortical pinwheel centres and iso-orientation domains. AB - Exposure to a high-contrast visual stimulus causes adaptation, a psychophysical phenomenon that is quite selective for stimulus orientation. Its mechanism is largely cortical but the underlying circuitry is still not unambiguously resolved. It has been suggested that adaptation could be the result of integration of inputs from cells within a large local pool, effectively scaling their outputs with respect to local contrast. In this case, orientation selectivity of neuronal adaptation should depend on the location of neurons within the cortical map of orientation preference. We tested this hypothesis by quantifying adaptation to optimally oriented and to orthogonal-to-optimum gratings among neurons recorded either from iso-orientation domains or orientation pinwheel centres, as identified by optical imaging of cat visual cortex. We did not find a significant difference in adaptation characteristics for these two populations of cells, implying that these characteristics do not depend on the local functional architecture. Surprisingly, however, we additionally observed that under isoflurane (but not halothane) anaesthesia, most neurons exhibited adaptation by cross-oriented gratings, regardless of their location within the orientation map. It seems likely that, under isoflurane, inputs became visible that were masked by the commonly used, deeper halothane anaesthesia. For individual cells, the presence of these inputs was independent of their location within the cortical orientation map. PMID- 11906529 TI - Group III metabotropic glutamate receptors and D1-like and D2-like dopamine receptors interact in the rat nucleus accumbens to influence locomotor activity. AB - Evidence for functional interactions between metabotropic glutamate (mGlu) receptors and dopamine (DA) neurotransmission is now clearly established. In the present study, we investigated interactions between group III mGlu receptors and D1- and D2-like receptors in the nucleus accumbens (NAcc). Administration, into the NAcc, of the selective group III mGlu receptor agonist, AP4, resulted in an increase in locomotor activity, which was blocked by pretreatment with the group III mGlu receptor antagonist, MPPG. In addition, pretreatment with AP4 further blocked the increase in motor activity induced by the D1-like receptor agonist, SKF 38393, but potentiated the locomotor responses induced by either the D2-like receptor agonist, quinpirole, or coinfusion of SKF 38393 and quinpirole. MPPG reversed the effects of AP4 on the motor responses induced by D1-like and/or D2 like receptor activation. These results confirm that glutamate transmission may control DA-dependent locomotor function through mGlu receptors and further indicate that group III mGlu receptors oppose the behavioural response produced by D1-like receptor activation and favour those produced by D2-like receptor activation. PMID- 11906531 TI - Comparison of neuronal activity in the rostral supplementary and cingulate motor areas during a task with cognitive and motor demands. AB - A number of cortical motor areas have been identified on the medial wall of the hemisphere in monkeys. However, their specific role in motor control remains unclear. In this study, we sought to describe and compare the functional properties of the presupplementary (pre-SMA) and rostral cingulate (CMAr) motor areas in two monkeys performing a visually instructed, delayed, sequential movement. We recorded 134 task-related neurons in the pre-SMA and 149 in the CMAr. The main difference between the two areas was the abundance of responses to targets (46%) in the pre-SMA, while CMAr activity was more related to reward (28%). Neuronal responses to targets were more phasic and higher in frequency in the pre-SMA than in the CMAr. During the delay, the percentage of neuronal responses was similar in the two areas. The discharge pattern was different depending upon whether the delay duration was fixed or variable but in most neurons was the same regardless of the sequence performed. Movement-related changes were common in the pre-SMA (75%) and in the CMAr (81%) but they occurred earlier in the former. Neurons activated exclusively during movement were more numerous in the CMAr. Finally, neuronal activity in the pre-SMA was more related to the sequential aspect of the task compared to the CMAr. Our results suggest that although the two areas share functional properties, they also participate in different aspects of motor behaviour. Their functional properties reflect their anatomical positions, which give them the potential to integrate external stimuli (pre-SMA) and internal states (CMAr) during motor planning. PMID- 11906532 TI - Basolateral amygdala lesions block the memory-enhancing effect of 8-Br-cAMP infused into the entorhinal cortex of rats after training. AB - There is extensive evidence suggesting that the basolateral nucleus of the amygdala plays a critical role in modulating memory consolidation processes in other brain regions. The present experiments examined interactions between the basolateral amygdala and the entorhinal cortex in modulating memory consolidation for inhibitory avoidance training. Several studies have reported that activation of the second messenger system adenosine 3',5'-cyclic monophosphate (cAMP) in several brain regions enhances memory and induces long-term plasticity. In the present experiments, a unilateral infusion of the cAMP analogue, 8-Br-cAMP (0.25 or 1.25 microg in 0.5 microL), administered into the entorhinal cortex of male Sprague-Dawley rats immediately after training, enhanced 48-h retention. An N methyl-d-aspartate-induced lesion of the ipsilateral basolateral amygdala did not impair retention, but blocked the memory-enhancing effect of 8-Br-cAMP (infused into the entorhinal cortex) post-training. A lesion of the contralateral basolateral amygdala did not block the 8-Br-cAMP-induced retention enhancement. These findings indicate that an intact basolateral amygdala is essential for modulation of memory consolidation involving the entorhinal cortex, and are consistent with evidence that the basolateral amygdala regulates memory consolidation mediated by other brain regions. PMID- 11906533 TI - A combined pharmacological and genetic approach to investigate the role of orphanin FQ in learning and memory. AB - Using a combination of the selective opioid receptor-like1 (ORL1) receptor agonist, Ro 64-6198, and orphanin FQ/nociceptin (OFQ/N) peptide knockout (KO) mice, the influence of OFQ/N on cognition has been studied in the rodent. In wild type, C57BL/6J mice, Ro 64-6198 (0.3-1 mg/kg i.p.) impaired the acquisition of spatial learning in the Morris water maze, although a mild neurological impairment was evident which complicated precise interpretation. In Lister hooded rats, Ro 64-6198 (6 mg/kg i.p.) produced delay dependent impairments in rats performing either a delayed matching or a delayed nonmatching to position task with only a modest (< 20%) effect on omissions - an effect consistent with a short-term memory impairment. Electrophysiological studies demonstrated an inhibitory effect of OFQ/N on LTP recorded from the CA1 region of wild type mice, but not in ORL1 receptor knockout mice. In contrast to the ORL1 agonist, mice deficient in the OFQ/N peptide showed some evidence of improved spatial learning, fear conditioning and passive avoidance retention. However, CA1 LTP was similar between OFQ/N peptide KO mice and wild type controls. Subsequent receptor radioautography studies demonstrated the presence of ORL1 receptors within various regions of the medial temporal lobe system: i.e. CA1, dentate gyrus molecular layer, subiculum, perirhinal cortex. Taken together, these results suggest a bi-directional effect of OFQ/N containing systems on aspects of cognitive behaviour, particularly those elements associated with hippocampal function. This is consistent with a likely modulatory role of OFQ/N on hippocampal and associated cortical circuitry. PMID- 11906534 TI - EEG theta synchronization conjoined with alpha desynchronization indicate intentional encoding. AB - The involvement of different oscillating neuronal systems activated during intentional learning was investigated by measuring ongoing EEG activity. In 17 subjects, the EEG was recorded while learning pairs of words and faces. Subjective task difficulty was rated and a control condition of mental relaxation was also run. Spontaneous EEG activity during epochs which subsequently resulted in efficient encoding was associated with upper alpha desynchronization (10-12 Hz) and theta synchronization (4-8 Hz) when compared with spontaneous EEG activity during epochs of poor recall performance. The combined measure of theta synchronization plus upper alpha desynchronization was enhanced selectively over left frontotemporal cortical regions during efficient learning of words and over right parietal cortical regions during efficient learning of faces (P < 0.001). This striking topographical dissociation between learning materials for the combined measure of theta and upper alpha EEG activity suggests that the mode of intentional learning relies essentially on an interdependent regulation of two neuronal circuits: the thalamo-cortical circuit and the hippocampo-cortical circuit. PMID- 11906535 TI - Expression of RGS2, RGS4 and RGS7 in the developing postnatal brain. AB - The abundant expression of RGS (regulator of G-protein signalling) proteins in neurons, together with their modulatory function on G-protein-dependent neurotransmission, provides the basis for cellular adaptation to sensory inputs. To identify the molecular mechanism involved in the sensory experience-induced neural development, we performed a systematic survey of the localization of mRNAs encoding three subtypes of the RGSs (RGS2, RGS4 and RGS7) in developing rat brains by in situ hybridization through postnatal day 2 (P2), P10 and P18 to adult. The most dramatic changes of expression patterns were observed in the discrete neuronal cell layers of the cerebral neocortex (for RGS2 and 4), the hippocampus (for RGS2, 4 and 7), the thalamus (for RGS4) and the cerebellum (for RGS2 and 7). In the neocortex, RGS2 mRNA was enriched in the superficial cortical plate at P2, in contrast to RGS4, which was enriched in more mature neurons of the deeper layer V and VI. In the hippocampus, the neuronal cell layer-specific expression pattern of RGS2 developed from P2 to P18. RGS4 expression was temporarily confined to the CA pyramidal cell layer and not detectable in the dentate gyrus at P10 and P18. Similarly, a high level of expression of RGS7 was observed in the CA area, but not in the dentate gyrus at P2 and P10. In the cerebellum, the maturation of laminar expression patterns for the three RGSs correlated with neuronal maturation and synaptogenesis at P18. The most characteristic temporal pattern among the three RGSs was observed for RGS4 mRNA, which was highly enriched in the thalamocortical regions. The peaks of RGS4 expression were seen in the following regions with distinct onset and duration: the neocortex (from P2 onward), the hippocampus (P10 and P18) and the thalamus (from P18 onward). The divergent temporal and spatial expression of RGS subtypes and their dynamic control in the cortex, the hippocampus and the thalamus suggest that the RGS family could play multiple distinct roles in experience-dependent brain development. PMID- 11906536 TI - Oestrogen-dependent tracing in the rat CNS after pseudorabies virus infection. AB - This study examines the hypothesis that neuronal infectivity and the spreading of the pseudorabies virus (PRV) through the synapses in the central nervous system (CNS) are influenced by the oestrogen levels. The arcuate nucleus (ARC) and the subfornical organ (SFO) were chosen as models for analysis; the neurons in both structures possess oestrogen receptors and are mutually connected. A genetically engineered pseudorabies virus (Ba-DupLac) was used as a transneuronal tract tracer. This virus is taken up preferably by axon terminals, and transported very specifically through the synapses in a retrograde manner. Ba-DupLac was injected into the ARC of rats, followed by monitoring of the PRV-immunoreactivity (PRV-IR) in the SFO 72 h following inoculation. We found no PRV immunolabelling in the SFO of ovariectomized (OVX) rats, or in those OVX animals that received oestrogen shortly (4 h) before PRV infection (OVX + E 4 h). In contrast, in those OVX animals that received oestrogen 12 h before PRV infection (OVX + E 12 h), and also in intact control animals, PRV-IR was demonstrated in the SFO in all cases. Surprisingly, a reverse labelling was observed in the OVX rats; PRV-IR appeared in the pyriform cortex, whereas PRV-IR could not be detected in the control and OVX + E 12 h animals. As far as we are aware, this is the first study to demonstrate that transneuronal PRV labelling depends on the effects of oestrogen on certain CNS structures and connections. PMID- 11906537 TI - Involving patients: representation or representativeness? PMID- 11906538 TI - Using disease risk estimates to guide risk factor interventions: field test of a patient workbook for self-assessing coronary risk. AB - OBJECTIVE: To assess the feasibility and acceptability of a patient workbook for self-assessing coronary risk. DESIGN: Pilot study, with post-study physician and patient interviews. SETTING AND SUBJECTS: Twenty southern Ontario family doctors and 40 patients for whom they would have used the workbook under normal practice conditions. INTERVENTIONS: The study involved convening two sequential groups of family physicians: the first (n=10) attended focus group meetings to help develop the workbook (using algorithms from the Framingham Heart Study); the second (n=20) used the workbook in practice with 40 patients. Follow-up interviews were by interviewer-administered questionnaire. MAIN OUTCOMES MEASURES: Physicians' and patients' opinions of the workbook's format, content, helpfulness, feasibility, and potential for broad application, as well as patients' perceived 10-year risk of a coronary event measured before and after using the workbook. RESULTS: It took an average of 18 minutes of physician time to use the workbook: roughly 7 minutes to introduce it to patients, and about 11 minutes to discuss the results. Assessments of the workbook were generally favourable. Most patients were able to complete it on their own (78%), felt they had learned something (80%) and were willing to recommend it to someone else (98%). Similarly, 19 of 20 physicians found it helpful and would use it in practice with an average of 18% of their patients (range: 1-80%). The workbook helped to correct misperceptions patients had about their personal risk of a coronary event over the next 10 years (pre-workbook (mean (SD) %): 35.2 (16.9) vs. post-workbook: 17.3 (13.5), P < 0.0001; estimate according to algorithm: 10.6 (7.6)). CONCLUSIONS: Given a simple tool, patients can and will assess their own risk of CHD. Such tools could help inform otherwise healthy individuals that their risk is increased, allowing them to make more informed decisions about their behaviours and treatment. PMID- 11906539 TI - Do Internet interventions for consumers cause more harm than good? A systematic review. AB - OBJECTIVE: To systematically review the effect of consumer use of online health information on decision-making, attitudes, knowledge, satisfaction and health outcomes and utilization. SEARCH STRATEGY: Electronic databases searched included the Cochrane Controlled Trials Register, MEDLINE, PREMEDLINE (to 14 March 2001), CINAHL, Australian Medical Index, Health and Society, National Institutes of Health Clinical Trials Database and CenterWatch. INCLUSION CRITERIA: All post 1995 comparative studies (including controlled studies, before and after studies, and interrupted time series analyses) of Internet users vs. non-Internet users and other communications mediums, and Internet characteristics such as e-mail vs. other communication mediums, were included. Outcomes included consumer decision making, attitudes, knowledge, satisfaction and measurable changes in health status or health utilization. DATA EXTRACTION AND SYNTHESIS: One reviewer screened all papers then two reviewers independently assessed studies against the selection criteria and any discrepancies were resolved by discussion with a third reviewer. No attempt was made to combine the data for further statistical analysis. MAIN RESULTS: We identified 10 comparative studies. Studies evaluated the effectiveness of using the Internet to deliver a smoking cessation programme, cardiac and nutrition educational programmes, behavioural interventions for headache and weight loss, and pharmacy and augmentative services. All studies showed some positive effects on health outcomes, although the methodological quality of many studies was poor. CONCLUSIONS: Despite widespread consumer Internet use to obtain health-care information, there is almost a complete lack of evidence of any effects this may have on health outcomes. PMID- 11906540 TI - Consequences of surveying folate awareness. AB - OBJECTIVES: To assess women's concerns when interviewed about the association between folate and neural tube defects (NTDs) and to determine how this is affected by time, being folate aware, having seen folate promotional material or being pregnant. DESIGN: As part of a community randomized trial outcomes evaluation, independent cross-sectional follow-up surveys were carried out in 1997 and 2000. SETTING AND PARTICIPANTS: Six local government areas in the state of Victoria, Australia; 2431 women aged between 15 and 44 years. MAIN VARIABLES STUDIED: Whether or not women knew of the association between folate and NTDs (i.e. were folate aware), whether or not women had been concerned by seeing folate/NTD information and if an interview about folate and NTDs had raised any concerns for them. RESULTS: In the 1997 survey, 36% of women said that the interview had raised concerns and this decreased to 26% in 2000. Women who were folate aware were significantly less likely to have raised concerns than women who were not folate aware (OR(adj) = 0.38, 95% CI 0.24-0.60). In general, women who had seen promotional material were less likely to feel concern about the interview than those who had not, although this varied with whether or not the promotional material had raised concerns. These effects were greater in women who were pregnant. CONCLUSIONS: Women had increased concerns having seen folate promotional material and after being interviewed about it. These results are consistent with the proposition that an initial emotional response to sensitive health information is part of an adaptive response appropriate to the process of health-related behaviour change. PMID- 11906541 TI - Deciding how NHS money is spent: a survey of general public and medical views. AB - OBJECTIVES: To examine the validity of the Prioritization Scoring Index (PSI) methodology by obtaining the views of our local population and clinicians regarding the criteria and weightings that should be used in deciding how NHS money is spent. BACKGROUND: We have used a PSI in Argyll and Clyde to allocate new money since 1996 and to determine priorities for our 1999/2000-2003/2004 Health Improvement Programme (HIP). Since the criteria and weightings for this methodology were developed subjectively, we sought to validate these by consulting local people and to change our methodology to take account of wider population views. METHODS: A postal questionnaire was sent to 1969 members of the general public, all 314 general practitioners and all 189 hospital consultants in Argyll and Clyde in March 1999. A reminder was sent after 4 weeks. Questions were asked about general funding and prioritization in the NHS and about specific issues relating to potential criteria for prioritization, including those used in our PSI methodology. Responses were analysed quantitatively in the Statistical Package for the Social Sciences (SPSS) and qualitatively through examination of the responses to open questions. RESULTS: The response rate was 51% for the general public and 71% for GPs and consultants. Respondents from the general public were broadly representative of the Argyll and Clyde population. The main findings were that: greater importance should be given to care that improves health, quality of life or prevents ill health rather than to cost, or to government and local health board priorities; half of the general public and most clinicians thought there should be a limit on NHS funding; extra money for the NHS should come from the national lottery (general public) or higher taxes on cigarettes and alcohol (clinicians); doctors should have the greatest influence in deciding how NHS money is spent; a higher priority should not be given to the health-care needs of younger people rather than older people. Our public and clinicians would allocate approximately 50% of the prioritization weighting to direct patient benefits, 25% to the cost of health-care and 25% to strategic health issues. CONCLUSIONS: Consideration of public and clinician views suggests that a revised PSI should place greater weight on benefits to patients and lower weight on the cost of health-care. PMID- 11906543 TI - Appraising the quality of consumer health information leaflets. PMID- 11906542 TI - Improving communication between health professionals and women in maternity care: a structured review. AB - OBJECTIVE: To review trials of the effectiveness of interventions aimed at improving communication between health professionals and women in maternity care. SEARCH STRATEGY: The electronic databases Medline, PsycLit, The Cochrane Library, BIDS Science and Social Science Indexes, Cinahl and Embase were searched. Final searches were carried out in April 2000. INCLUSION CRITERIA: Controlled trials of interventions explicitly aimed at improving communication between health professionals and women in maternity care were included. Other trials were included where two reviewers agreed that this was at least part of the aim. DATA EXTRACTION AND SYNTHESIS: 95 potentially eligible papers were identified, read by one reviewer and checked against the inclusion criteria. The 11 included trials were read, assessed for quality and summarized in a structured tabular form. RESULTS: The included trials evaluated interventions to improve the presentation of information about antenatal testing, to promote informed choice in maternity care, woman-held maternity records and computer-based history taking. Four trials in which women were provided with extra information about antenatal testing in a variety of formats suggested that this was valued by women and may reduce anxiety. Communication skills training for midwives and doctors improved their information giving about antenatal tests. The three trials of woman-held maternity records suggested that these increase women's involvement in and control over their care. CONCLUSIONS: The trials identified by this review addressed limited aspects of communication and focused solely on antenatal care. Further research is required in several areas, including trials of communication skills training for health professionals in maternity care and other interventions to improve communication during labour and in the postnatal period. PMID- 11906544 TI - The epidemiology of ovarian cancer. PMID- 11906545 TI - Clinical behavior of 203 stage II endometrial cancer cases: the impact of primary surgical approach and of adjuvant radiation therapy. AB - The aim of this study was to verify the impact of primary surgical approach and adjuvant radiation therapy (RT) on survival, recurrence rate, and pattern of relapse in stage II endometrial cancer patients. Two hundred three subjects were retrospectively reviewed; 135 (66%) underwent simple hysterectomy (SH) and 68 (34%) radical hysterectomy (RH). Sixty-six of 111 (59%) of stage IIA and 67 of 92 (73%) of stage IIB patients underwent adjuvant radiation therapy. Actuarial survival rates for stage IIA and IIB were 86% and 74% at 5 years and 82% and 68% at 10 years, respectively. Survival rates by surgical procedure were 79% in the SH group and 94% in the RH group at 5 years and 74% and 94% at 10 years, respectively (P < 0.05). The overall recurrence rate was 13.8% (28/203); by adjuvant treatment it was 18.6% (13/70) in the observation group and 11.3% (15/133) in the RT group. Most of the relapses were locoregional in the observation group and distant in the RT group. Survival rates by RT were not statistically different. Subjects treated with RH improved their survival compared with the SH group; the difference was significant, but randomized studies should confirm this trend. Although adjuvant RT seemed to reduce the recurrence rate, there was no significant difference in survival, and so the role of RT still needs further verification. PMID- 11906546 TI - A combination of platinum and tamoxifen in advanced ovarian cancer failing platinum-based chemotherapy: results of a Phase II study. AB - The treatment of recurrent or progressive ovarian cancer has limited therapeutic potential. The clinical outcome of second-line therapy largely depends on the potential chemo-sensitivity of the tumor expressed during up-front chemotherapy, as well as on the treatment-free interval from the last course of cytotoxic therapy. However, the identification of agents such as tamoxifen (TAM) at nontoxic doses, able to act synergistically with standard chemotherapy, may be useful to overcome resistance. Fifty patients with recurrent or progressive ovarian cancer following platinum (P)-based chemotherapy (28 platinum-resistant and 22 platinum-sensitive) entered a Phase II trial to evaluate the efficacy and toxicity of P re-challenge with the addition of TAM as a chemotherapy response modulator. The choice of the P compound (100 mg/m2 cisplatin or 400 mg/m2 carboplatin, q3 weeks) was made on the basis of the prior total cisplatin dose and the presence of neurotoxicity. TAM was administered at the doses of 80 mg/day for 30 days followed by 40 mg/day for the remaining period of treatment. Toxicity consisted mainly of mild to moderate nausea and vomiting (76%), peripheral neuropathy (43%), nephrotoxicity (4%), anemia (16%), leukopenia (58%) and thrombocytopenia (16%). The overall response to the P-TAM combination was 50% (complete response 30%; partial response 20%) with a median duration of 8.5 months (3-42). Sixty-four percent of the P-sensitive and 39% of the P-resistant patients responded (59% and 33%, respectively, for those bearing measurable disease). The overall median survival was 23 (3-48) and 19 months for the patients with measurable disease (20 months for the P-resistant group). This phase II trial confirmed the activity for a re-challenge employing a P compound and TAM in clinically defined P-resistant ovarian cancer patients. The mild toxicity profile and the relatively low cost of the treatment render further investigations on the P-TAM regimen worthwhile. PMID- 11906547 TI - High-dose-rate brachytherapy at 14 Gy per hour to point A: preliminary results of a prospectively designed schedule for cancer of the cervix based on the linear quadratic model. AB - The objective of this study was to describe the results and complications of a prospectively designed high-dose-rate (HDR) brachytherapy schedule for early stage cancer of the cervix, at 14 Gy/h to point A, based on the linear-quadratic model and our clinical experience. We used a combination of brachytherapy and external beam pelvic and parametrial irradiation in 88 consecutively seen patients with stage IB1-IIB treated by irradiation alone (1995-1998). The modeled HDR schedule consisted of three insertions on three treatment days separated by 10 days, with six 7 Gy planned brachytherapy fractions to point A, at 14 Gy/h, two on each treatment day with an interfraction interval of 6 h, plus an 18 Gy external whole-pelvic dose followed by additional parametrial irradiation. The calculated biologically effective dose (BED) was 92 Gy10 for tumor and 110 Gy3 for the rectum, equivalent to 77 and 66 Gy in 2 Gy fractions, respectively. The median overall treatment time was 41 days. The actuarial 4-year central recurrence-free rate, pelvic control, and disease-free survival rate were 97%, 93%, and 88% for stages IB-IIA and 79%, 75%, and 75% for stage IIB. The actuarial 4-year late complication rate for grades 2-3 was 4.7% (scale 0-3). We conclude that preliminary results of this HDR brachytherapy schedule for early-stage disease at a median follow-up of 52 months are as effective as the previously used low dose rate (LDR) at 0.44 Gy/h at point A. They are also as effective as medium-dose-rate schedules (MDR) at 1.6-1.5 Gy/h at this institution and do not require a further increase in fractionation of intracavitary treatments or in the whole-pelvic external beam irradiation dose common to standard HDR schedules. In addition, more patients per machine can be treated per day compared with MDR. Longer follow-up is required for a complete assessment of late complications. PMID- 11906548 TI - Expression of the protease inhibitor antileukoprotease and the serine protease stratum corneum chymotryptic enzyme (SCCE) is coordinated in ovarian tumors. AB - We have previously reported that the stratum corneum chymotryptic enzyme (SCCE) is overexpressed in ovarian cancers and that SCCE has potential as a useful marker and/or a therapeutic target for ovarian carcinoma. Antileukoprotease (ALP) has been shown to be a specific inhibitor of SCCE. The objective of this study was to investigate the potential cotranscription and overexpression of ALP in carcinoma of the ovary. The expression of ALP transcript was evaluated by Northern blot hybridization and by semiquantitative polymerase chain reaction (PCR) technique. The presence of the ALP protein in ovarian tumor cells was evaluated by immunohistochemistry. Northern blot hybridization showed that the ALP transcript was abundant in ovarian carcinomas but was not detected in the normal ovary. Semi-quantitative PCR examination revealed that the mRNA level of ALP was significantly elevated in low-malignant-potential tumors and in ovarian carcinomas compared with that in normal ovaries (P < 0.01). There was significant positive correlation between SCCE and ALP mRNA overexpression status in ovarian tumor cases (P < 0.01). Immunohistochemical expression of ALP protein was observed in ovarian tumor cells, whereas little or no staining was observed in normal ovarian surface epithelium. Like SCCE, ALP is highly overexpressed in ovarian tumor cells, which begs the question of whether it remains an effective inhibitor of SCCE or whether it is discordant in time or space and is ineffective as an inhibitor of the SCCE enzyme. PMID- 11906549 TI - c-myc gene amplification detected in preinvasive intraepithelial cervical lesions. AB - The aim of the present work was to evaluate the c-myc gene amplification process in cervical samples and to analyze the relationship between the activation of this proto-oncogene and the cytologic and/or histologic status. Thirty-four normal cervical samples and 105 abnormal cervical tissue scrapes, previously used for PAP or histopathologic diagnosis, were analyzed for c-myc gene amplification. Detection of c-myc gene amplification was performed using a polymerase chain reaction (PCR)-based method known as target arbitrarily primed-PCR (TAP-PCR). For c-myc amplification, significant differences were found between normal samples and samples presenting different grades of lesions (P<0.001). A significant difference between high-grade squamous intraepithelial lesions (HG-SIL) and the other stages of cervical disease was also found (P<0.05). This study demonstrated that c-myc copy number increases according to the histologic grade of the lesion. These results could indicate that oncogene amplification takes place in preinvasive stages of cervical disease and could cooperate not only in tumor progression but also in cell transformation. PMID- 11906550 TI - Neoadjuvant chemotherapy or primary surgery in advanced epithelial ovarian carcinoma. AB - Neoadjuvant chemotherapy has been proposed as an alternative approach to conventional surgery as initial management of bulky ovarian cancer, with the goal of performing adequate debulking in the interval surgery. Two hundred five consecutive patients with advanced ovarian cancer were divided into two groups. Neoadjuvant chemotherapy followed by interval surgery was performed in 45 of 205 patients. The remaining 158 patients received primary surgery plus adjuvant chemotherapy. Optimal cytoreductive surgery rates were significantly higher in the neoadjuvant CT group (P<0.001). In multivariate analysis, only residual tumor diameter and appendix involvement were found to affect total survival significantly in both groups. Five-year survival and median survival were not statistically different when all patients treated conventionally were compared with all patients treated with neoadjuvant chemotherapy. Primary chemotherapy followed by interval debulking surgery in a selected group of patients does not appear to worsen prognosis, but it permits less aggressive surgery and improves patients' quality of life. PMID- 11906551 TI - Omentectomy, peritoneal biopsy and appendectomy in patients with clinical stage I endometrial carcinoma. AB - The aim of this study was to evaluate whether omentectomy, appendectomy, and peritoneal biopsy should be a routine part of staging surgery in endometrial carcinoma. Data of 97 patients who had been diagnosed with clinical stage I endometrial carcinoma were reviewed. Associations in the data obtained, pelvic and para-aortic lymph node status, depth of myometrial invasion, grade, and histology were investigated. The chi-square (chi2) test was used for statistical analysis. Of 97 patients, six (6%) had omental metastases, which was microscopic in four. There was a statistically significant relationship between omental metastasis and tumor grade (P < 0.01). Deep myometrial invasion was significantly more common in patients with omental metastases. Tumor was found in one of 55 appendectomy specimens (2%). Omentectomy may be included in surgical staging in patients with deeply invasive or grade 3 endometrial cancer because of the possibility of omental metastasis in spite of what appears to be stage I disease in laparotomy. In other cases, omentectomy and appendectomy and biopsies from peritoneal sites should be performed in the presence of grossly suspicious disease. PMID- 11906552 TI - Postoperative adjuvant therapy in early invasive cervical cancer patients with histopathologic high-risk factors. AB - The purpose of this study is to evaluate the efficacy of postoperative adjuvant therapy in preventing treatment failure after primary treatment with surgery in early invasive cervical cancer patients associated with the following histopathologic high-risk factors: lymph node metastasis (either macroscopic or microscopic), parametrial extension, lymphovascular permeation and depth of invasion > or =10 mm. Postoperative adjuvant concurrent chemoradiotherapy (PCCRT), postoperative adjuvant chemotherapy (PCT), or postoperative adjuvant radiotherapy (PRT) alone was administered to the 80 early invasive cervical cancers with at least one of the high-risk factors. Each of 61 patients received three to six cycles of chemotherapy at intervals of approximately 3 weeks. Twenty three patients were treated with PCCRT, 38 patients were treated with PCT alone, and 19 patients received PRT. The 5-year survival rates of patients with macroscopic lymph node metastasis were 66.7% and 35.7% in PCCRT and PRT, respectively. With microscopic lymph node metastasis, the 5-year survival rates were 83.3%, 60.0%, and 70.1% in PCCRT, PCT, and PRT, respectively. With parametrial extension, the 5-year survival rate was 58.1% in PCCRT. The 5-year survival rates of patients with lymphovascular permeation were 100%, 90.9%, and 66.7% in PCCRT, PCT, and PRT, respectively. With depth of invasion > or =10 mm, the 5-year survival rates were 100% and 91.3%, in PCCRT and PCT, respectively. PCCRT appears to be superior to PRT or PCT alone in early invasive cervical cancer patients with histopathologic high-risk factors. PMID- 11906553 TI - Association between menopausal state and prognosis of endometrial cancer. AB - The purpose of our study was to examine the menopausal state as an independent prognostic variable of endometrial cancer and determine the conditions under which the menopausal state would be an independent prognostic variable of such cancer. We studied the clinical and pathologic variables of 255 patients with invasive endometrial cancer. In comparisons of the clinicopathologic variables between menopausal states, obesity and deep myometrial invasion were found more frequently in older patients than in younger ones. Multivariate analysis performed on 255 cases with complete pathologic data identified menopausal state, cervical invasion, pelvic lymph node metastasis, and tumor grade as prognostic variables. Univariate analysis revealed that survival of older patients with FIGO pathologic stage Ib disease was significantly poorer than in younger patients, while there was no significant difference in the analyses of stage Ic or advanced disease beyond stage II. We conclude that menopausal state was an independent prognostic variable for patients with early endometrial cancer, but not for patients with advanced disease. PMID- 11906555 TI - Cervical schistosomiasis. AB - We present a case history of a woman who was diagnosed as having cervical schistosomiasis on histology following investigations for abnormal cervical smear. Schistosomiasis of the female genital tract can present with varied symptoms and there is a need for greater awareness of this diagnosis as the number of travellers to schistosomiasis-endemic areas rises. Travellers to these areas should be warned about the risk of swimming in lakes and rivers. PMID- 11906554 TI - Endometrial carcinoma coexisting with pregnancy, presumed to derive from adenomyosis: a case report. AB - Endometrial carcinoma coexisting with pregnancy is rarely observed. We report here the case of a 35-year-old woman with an endometrial carcinoma that was diagnosed 6 months after childbirth. Preoperative magnetic resonance imaging (MRI) revealed a cystic mass attached to the uterus, with a papillary projection on the wall of the mass. The patient underwent complete surgical extirpation and five postoperative courses of adjuvant chemotherapy, given that the tumor contents had leaked into the peritoneal cavity when the capsule of the tumor ruptured intraoperatively. Microscopic examination revealed an endometrioid adenocarcinoma in the muscular layer close to the uterine serosa that was presumed to derive from adenomyosis. Further investigation is required to elucidate the pathogenesis of endometrial carcinoma in association with pregnancy and adenomyosis. PMID- 11906556 TI - Invasive adenocarcinoma of the vagina following surgery for adenocarcinoma in situ of the cervix--recurrence or implantation? AB - A 51-year-old woman underwent cervical conization for severe glandular abnormal cells. Histology noted adenocarcinoma in situ (AIS) with incomplete excision margins. Four months later, hysterectomy revealed no residual disease. Six months subsequently she developed invasive adenocarcinoma of the upper vagina. This report documents the unusual behavior of AIS and its management difficulties. PMID- 11906557 TI - Effects of vitamin C and vitamin E on performance, digestion of nutrients and carcass characteristics of Japanese quails reared under chronic heat stress (34 degrees C). AB - This study was conducted to determine the effects of vitamin C (L-ascorbic acid) and vitamin E (DL-alpha-Tocopheryl acetate) on performance, digestion of nutrients and carcass characteristics of Japanese quails reared under chronic heat stress (34 degrees C). A total of 180 10-day-old Japanese quails were randomly assigned to six treatment groups, three replicates of 10 birds each. The birds with a 2 x 3 factorial design received either two levels of vitamin C (100 and 200 mg/kg of diet) or three levels of vitamin E (125, 250, or 500 mg/kg of diet). Then, 200-mg vitamin C/kg of diet, compared with that of 100 mg/kg of diet, and higher dietary vitamin E inclusions resulted in a higher performance. The interaction between vitamin C and vitamin E for final body weight change (p=0.01) and feed efficiency (p=0.02) was detected. Final body weight change and feed efficiency increased to a higher extent by increasing dietary vitamin C when higher vitamin E levels were fed. Carcass characteristics improved with an increase of both dietary vitamin C and vitamin E (p=0.004). The interactions on carcass characteristics were all significant (p=0.02) and manifested themselves in a way that they were improved to a higher extent by an increase of dietary vitamin C when higher vitamin E levels were fed. Digestibility of nutrients (DM, OM, CP and EE) was greater with higher dietary vitamin C (p < 0.02) and also with higher vitamin E (p=0.07). There were no interactions detected for digestibility of nutrients (p=0.32). Taken together, the results of the present study conclude that a combination of 200 mg of vitamin C and 250 mg of vitamin E provides the greatest performance in Japanese quails reared under heat stress and can be considered as a protective management practice in poultry diet, alleviating the negative effects of heat stress. PMID- 11906558 TI - Effects of vitamin E and selenium on performance, digestibility of nutrients, and carcass characteristics of Japanese quails reared under heat stress (34 degrees C). AB - This study was conducted to determine the effects of vitamin E (dL-alpha Tocopheryl acetate) and selenium (Se; Na2-SeO3) on performance, digestibility of nutrients and carcass characteristics of Japanese quails reared under chronic heat stress (34 degrees C). A total of 120 10-day-old Japanese quails were randomly assigned to four treatment groups, three replicates of 10 birds each. The birds with a 2 x 2 factorial design received either two levels of vitamin E (125 and 250 mg/kg of diet) or two levels of Se (0.1 or 0.2 mg/kg of diet). A 250 mg vitamin E/kg of diet compared with that of 125 mg/kg of diet and higher dietary Se inclusions (0.1 vs. 0.2 mg/kg) resulted in a better performance (p=0.001). The interaction between vitamin E and Se for feed intake (p=0.03), final body weight change (p=0.03) and feed efficiency (p=0.001) was detected. Carcass yield increased with increasing both dietary vitamin E and Se (p=0.001). The interactions on carcass characteristics were all non-significant (p > 0.06). Digestibility of nutrients (DM, OM, CP and ether extract) was higher with higher dietary vitamin E (p=0.03), and DM digestibility was also higher with higher dietary Se (p=0.05). There were no interactions detected for digestibility of nutrients (p=0.28). From the results of the present study, it was concluded that a combination of 250 mg of vitamin E and 0.2 mg of Se provides the greatest performance in Japanese quails reared under heat stress and this combination can be considered as a protective management practice in Japanese quail diets, reducing the negative effects of heat stress. PMID- 11906559 TI - Effects of dietary protein level and cold exposure on tissue responsiveness and sensitivity to insulin in sheep. AB - The effects of dietary crude protein (CP) level and cold exposure on tissue responsiveness and sensitivity to insulin were studied in sheep. Nine rams were assigned to one of three isoenergetic diets which contained 70, 100, and 140% of CP for maintenance. They were exposed from a thermoneutral environment (20 degrees C) to a cold environment (0 degrees C) for 7 days. A hyperinsulinemic euglycemic clamp approach was applied for the determination of tissue responsiveness to insulin (the maximal glucose infusion rate, GIRmax) and tissue sensitivity to insulin (the plasma insulin concentration at half maximal glucose infusion rate, ED50). Dietary CP level influenced digestibilities of dry matter and CP (P=0.002 and P=0.001, respectively), and cold exposure decreased (P=0.01) CP digestibility. The GIRmax and ED50 tended to be influenced (P=0.08) by dietary CP level. The GIRmax was enhanced (P=0.0001) during cold exposure. Significant interactions between diet and environment were found for the GIRmax (P=0.04), but not for ED50 (P=0.07). It is concluded that in sheep dietary CP level can modify insulin action in response to cold exposure. PMID- 11906560 TI - Study of the optimum ideal protein level for weaned piglets. AB - The amount of ideal protein, represented by the first limiting valine (Val), for optimising the growth performances of weaned hybrid piglets was studied. Feeds were formulated based on the ideal protein concept and on a constant essential/nonessential amino acid (AA) ratio and net energy (NE) level. The animal performance trial was composed of five dietary treatments ranging from 0.57 to 0.81% calculated apparent (app.) ileal (il.) digestible (dig.) Val. The piglets, male and female in equal numbers (11 replicates x 6 piglets/pen x 5 treatments) entered the trial at about 4 weeks old (average live weight 8.1 kg). The piglets were a cross product of Pietrain sire x hybrid dam. Feed intake and weight were recorded every two weeks until the end of the trial at 10 weeks of age (average live weight 20.6 kg). The requirement was expressed in st. il. dig. AA-units, as this unit approaches available AA better than app. il. dig. AA. The standardised (st.) il. AA digestibility coefficients (DCAA) were determined for two feeds, close to the animal performance optima, in a digestion trial with four T-cannulated piglets of approximately 6 weeks old. The feed independent endogenous nitrogen excretion was measured with a protein-free feed; although this technique underestimates the actual endogenous N-losses, it provides a reasonable estimate of basal endogenous N-losses. The determined st. il. DCAA were lower than the calculated st. il. DCAA, based on the CVB (2000); this might be linked to the higher than expected crude fibre content of the experimental feeds. The Val-requirement necessary to optimise ADG and FCR was similar and amounted 0.70% st. il. dig. Val, which corresponded with a st. il. dig. Lys-level of 1.03%. PMID- 11906561 TI - Application of the E-screen assay to test for oestrogenically active substances in swine feed. AB - A pig breeder in central Hesse (Germany) noticed the occurrence of enlarged vulvae in female piglets. Intoxication with oestrogenically active substances by contamination of two feed mixes ingested by the mother sows appeared to be a possible cause. Using a combined technique of the DFG analytical method S19 and the E-screen assay, two feed samples were found to contain powerful oestrogenically active compounds. By co-incubation with the anti-oestrogen tamoxifen it could be clearly demonstrated that the oestrogenic activity was mediated by the oestrogen receptor. These results demonstrate that use of the E screen assay in combination with the DFG analytical method S19 provides a simple and readily usable prescreening method for the routine detection of oestrogenically active compounds in animal feed. The results from the E-screen assay show that the sows ingested 10-80 microg oestradiol equivalents per day in their feed. Because of the bioavailability of these substances, the oestrogenic active compounds seem to be transferred into the milk and passed to the piglets via suckling. The milk of the dam appears to contain this substance in biologically active form and at such high concentrations that the female piglets had enlarged vulvae. PMID- 11906562 TI - Effect of plant oils and aspartate on rumen fermentation in vitro. AB - The effect of plant oils and aspartate (ASP) on rumen fermentation in vitro. The objective of this study was to determine the effect of plant oils (rapeseed - RO, sunflower - SO; linseed - LO; 10% wt/wt) and 8 mmol sodium aspartate on rumen fermentation of a diet (250 mg) consisting of hay, barley and sugar beet molasses (60 : 30 : 10). Rumen fluid was collected from two Slovak Merino sheep fed the same diet twice daily. Mixed ruminal micro-organisms were incubated in fermentation fluid (40 ml) containing rumen fluid and McDougall's buffer (1 : 4). Incubations were carried out in batch cultures for 72 h at 39 degrees C two times in a 3-week intervals. When compared to the control, all supplemented diets (RO, SO, LO, ASP) significantly increased the pH, the mol% of propionate (LO + 8.7%; SO + 10.12%; RO + 8.65%; ASP + 5.86%) and the acetate : propionate ratio and numerically decreased methane production (SO -32.8%; LO, RO -30.08%; ASP 21.56%). Lactate production was also significantly decreased. Addition of plant oils to aspartate-treated incubations partly inhibited the decrease of n butyrate, lactate and the increase of pH and in vitro dry matter digestibility (IVDMD) caused by ASP treatment. The effect of combined additives (RO + ASP, SO + ASP, LO + ASP) on methane production SO + ASP (-19.23%) and mol% propionate SO + ASP (+2.66%), LO + ASP (+4.28%) was less effective. All combined additives caused a significant decrease in digestibility of the given feeds. No effect of plant oils and ASP could be observed on the parameters of rumen fermentation (mainly methane and propionate). PMID- 11906563 TI - [The effects of the primary growth of two ryegrasses on the dynamics of changes in digestibility and feed intake by sheep]. AB - Dynamics of changes in digestibility and feed intake by sheep of two ryegrass species during primary growth Environmental impacts can cause short-term variations in chemical composition and feeding value of grass swards during growth and maturation and induce concomitant changes of intake of these grasses by animals. Continuous digestion trials are effective in observing the dynamics of these variations. Therefore this study was conducted to evaluate the effects of maturity-related alterations in the chemical composition of two ryegrass swards during primary growth on nutrient digestibilities and feed intake by sheep. Pure swards of perennial ryegrass (Lolium perenne L., variety Gremie) and Italian ryegrass (Lolium multiflorum Lam., variety Lema) were harvested daily during primary spring growth between 23 April and 19 June 1991 and each grass was offered to a group of five sheep for ad libitum intake. A continuous digestion trial design was employed to measure daily intakes and faecal outputs of organic matter (OM) and OM constituents. Digestibility values were calculated with the assumption that faecal output of day 2 represented the undigested nutrients of the grass eaten at day 0. Based on results of a preliminary study, daily intakes and digestibilities were expressed as rolling 3-day averages. The chemical composition of the grasses varied considerably with growth. Crude protein concentrations declined from 18.3 to 5.9% of dry matter (DM) for perennial ryegrass and from 17.0 to 5.4% of DM for Italian ryegrass, whereas concentrations of some cell-wall fractions markedly increased and partly more than doubled. Fibre content was slightly greater for perennial ryegrass than for Italian ryegrass over the whole growth period. The decline of feeding value with maturity was closely related to increasing concentrations of fibre components and their declining digestibility. Intake by sheep of Italian ryegrass was higher by 3 g/kg of body weight at the beginning and at the end of the primary growth period. Digestibility of the OM was greater than 80% at the start of the experiment and still around 70% after 2 months of grass growth. The decline in DM and metabolizable energy intake was much more pronounced than the decrease of OM digestibility. This could be attributed in part to a decline in feed intake capacity of non-lactating sheep caused by increasing body fat mass. Short-term fluctuations in OM digestibilities were related to alterations of chemical composition of the grasses caused by growth and maturation, in particular variations in water-soluble carbohydrate concentrations. The two ryegrass species differed with regard to nitrogen associated with the cell-wall (NDF) and lignocellulose (ADF). Continuous digestion trials were effective in observing the dynamics of alterations in feeding value and feed intake by sheep as related to growth and maturation of two ryegrass species during primary spring growth. PMID- 11906564 TI - Hydrolysis of phytic acid by intrinsic plant or supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re entrant cannulas. AB - Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (A. niger) were investigated with six minipigs (40-50 kg initial BW) fitted with re-entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5, and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2, and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn based diet (43 U Phytase/kg DM); (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM) and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 0730 and 1930 per animal 350 g diet mixed with 1050 ml de-ionized water were fed. Digesta were collected continuously and completely during 12 h after feeding. Duodenal recovery of dry matter and total phosphorus were 100% in the period between two feedings, irrespective of dietary treatment. In animals fed the wheat based diet, dry matter left the stomach faster (p < 0.05) during the first hour after feeding than in animals fed the corn-based diets (41.3 vs. 31.0 and 25.8% of intake, respectively). Supplemented microbial phytase did not affect ileal dry matter digestibility of the corn-based diet. In the first hour after feeding, phosphorus concentration of the duodenal digesta of animals fed corn-based diets with or without supplemented microbial phytase (5.86, 6.19 mg total P/g DM) exceeded the dietary level considerably (4.30 and 4.21 mg total P/g DM) indicating a higher solubility of corn than wheat phosphorus in the stomach. Apparent ileal P absorption was higher (p < 0.05) in the wheat-based diet (37.6%) and corn-based diet supplemented with microbial phytase (34.3%) than in the unsupplemented corn-based diet (17.6%). PMID- 11906565 TI - Hydrolysis of phytic acid by intrinsic plant and supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re entrant cannulas. 2. Phytase activity. AB - Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (A. niger) were investigated with six minipigs (40-50 kg initial BW) fitted with re-entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5, and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2, and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn based diet (43 U phytase/kg DM); (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM) and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 0730 and 1930 per animal 350 g diet mixed with 1050 ml de-ionized water were fed. Digesta were collected continuously and completely during 12 h after feeding. In the duodenal digesta, 70% of the microbial phytase (diet 2) and 45% of the wheat phytase (diet 3) were recovered within 12 h after ingestion of the phytases, whereas only negligible amounts were detected in the digesta of pigs fed the phytase-poor corn-based diet 1. Most phytase activity passed through the stomach within the first hour after feeding. Microbial phytase activity at pH 2.8 was less sensitive to acidic pHs, such as those found in the stomach, than phytase activity at pH 5.3. Phytase activities in the digesta of the distal ileum did not depend either on source or amount of dietary phytase activity. PMID- 11906566 TI - Hydrolysis of phytic acid by intrinsic plant and supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re entrant cannulas. 3. Hydrolysis of phytic acid (IP6) and occurrence of hydrolysis products (IP5, IP4, IP3 and IP2). AB - Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (Aspergillus niger) were investigated with six minipigs (40-50 kg initial body weight) fitted with re entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5 and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2 and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet [43 U phytase/kg dry matter (DM)]; (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM); and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 07 30 h and 19 30 h, each animal was fed 350 g diet mixed with 1050 ml de-ionized water. Digesta were collected continuously and completely during a 12-h period after feeding. Mean hydrolysis rates of IP6 in the stomach as measured at the proximal duodenum of animals 1, 4, 5 and 6 were 9.0, 77.2 and 66.2% for diet 1, 2 and 3, respectively. Microbial phytase was much more effective in phytate hydrolysis than wheat phytase. Mean IP6 hydrolysis rates of the respective diets in the stomach and small intestine as measured at the distal ileum of animals 1, 2 and 3 were 19.0, 62.6 and 64.6% and were lower than treatment means of the stomach only. Differences existed between experimental animals with respect to their ability to hydrolyse IP6 in the stomach independent of the presence and source of dietary phytase. Considerable amounts of hydrolysis products occurred in both the duodenal and ileal digesta when diets 2 and 3 were fed; however, only traces were determined after ingestion of diet 1. Independent of dietary treatment, four IP5 isomers were detected, but in different amounts. PMID- 11906568 TI - Influence of dietary phosphorus deficiency with or without addition of fumaric acid to a diet in pigs on bone parameters. AB - The purpose of this study was to examine if substantial bone loss occurs in weaned pigs by feeding a phosphorus-deficient diet with or without fumaric acid. Eighteen weaned pigs were used. The animals were assigned to three groups: group C (control; 0.65% P on DM basis), group LP (low phosphorus; 0.37% P on DM basis) and group LPF (low phosphorus plus fumaric acid; 0.35% P on DM basis plus 2% fumaric acid). These three diets were fed to the groups for a period of four weeks after a two-week adaptation period. Blood samples were collected once a week. Carboxyterminal telopeptide of type I collagen (ICTP) in serum was used as a bone resorption marker. Osteocalcin (OC) and bone-specific alkaline phosphatase (bAP) were used as bone formation markers. Bone mineral density (BMD) and content (BMC) were determined by peripheral quantitative computer tomography. BAP activities significantly increased (24%) in group LPF, and at the last sampling day group LPF had significantly increased activities in comparison to group C. In contrast, ICTP concentrations significantly increased with time in group LP and LPF, and at the last sampling day group LPF had significantly increased activities in comparison to group C. BMD and BMC in femur and tibia significantly decreased in group LP and LPF. The results show that P-deficient diets induce a bone loss. Fumaric acid did not influence the degree of bone loss. With a better understanding of its effect on bone, dietary phosphorus requirements in pigs could be more precisely defined. PMID- 11906569 TI - Growth performance and health status in weanling piglets fed spray-dried porcine plasma under typical Northern European conditions. AB - The effect of inclusion of spray-dried porcine plasma (SDPP) in diets for weanling piglets was studied. The objectives were to determine whether SDPP would have positive effects on post-weaning piglet performance and health under typical Northern European conditions. In experiment 1, 160 weanling piglets were assigned randomly to a control diet or a diet containing 3% SDPP, which was added at the expense of both fishmeal and dried skim milk. In experiment 2, 264 weanling piglets were assigned to a control diet containing whey protein, a diet without whey protein but with SDPP or a diet containing both whey protein and SDPP. In essence, SDPP was added to the test diets at the expense of either whey protein or fishmeal. Piglets were fed the diets for 3 weeks. In experiment 1, the piglets fed the SDPP diet had a 7% higher average daily gain (ADG) and a 4% lower feed conversion ratio (FCR) (p < 0.05) during the first 3 weeks after weaning than did those fed the control diet. There were no differences in leucocyte counts or gamma-globulin. In experiment 2 there were no significant differences in ADG and FCR among the dietary treatments. It is concluded that low amounts of SDPP in weanling diets can have positive effects on growth performance under Northern European conditions. PMID- 11906570 TI - [The feed value in growing pigs of a new cultivar of field beans (Vicia faba L.) supplemented with DL-methionine or DL-methionine-hydroxyanalog]. AB - A basal control mixture of barley, soy bean meal and soy bean oil was replaced by 25% of the new field bean-cultivar 'Divine' and the resulting two mixtures were supplemented with minerals, trace elements, vitamins and amino acids according to the ideal protein concept. The control diet was adjusted with DL-methionine (DL Met), the field bean mixture either with DL-Met or DL-methionine-hydroxyanalogue (DL-MHA) assuming biological equivalence on a molar basis for both supplements. The three experimental diets were fed to growing pigs (35-40 kg bwt.). Spontaneous urine samples were analysed separately for determining parameters that characterize the acid-base status of the pigs. There were no significant differences between experimental groups in nutrient digestibilities. The level of bacterially fermentable substances was increased in the diets containing field beans. The field beans contained 14 mg ME/kg DM. There were no significant (p < 0,05) differences in N- and mineral-retentions (Ca, P, Na, K) between the treatments. The stronger alkalinity found in urine after feeding the field bean mixtures resulted from a higher electrolyte balance of the diet. PMID- 11906571 TI - A low-selenium diet increases thyroxine and decreases 3,5,3'triiodothyronine in the plasma of kittens. AB - The effect of a low-selenium diet on thyroid hormone metabolism was investigated in growing kittens. Twelve specific-pathogen-free kittens with ages ranging from 16 to 18 weeks were divided into two groups of equal number with equal sex distribution in each group. One group was fed a yeast-based low-selenium diet (0.02 mg Se/kg diet) while the other group was fed the same diet supplemented with Na2SeO3 at 0.4 mg Se/kg diet for 8 weeks. Food intake, body weight and body weight gain were not affected by the low-Se diet during the study period. However, kittens given the low-Se diet had significantly reduced plasma selenium concentration and glutathione peroxidase activity. Plasma total thyroxine (T4) increased and total 3,5,3'triiodothyronine (T3) decreased significantly in kittens fed the low-Se diet at the end of the study. These results suggest that type I deiodinase in cats is a selenoprotein- or a selenium-dependent enzyme. PMID- 11906572 TI - Influence of the probiotic strain Bacillus cereus var. toyoi on the development of enterobacterial growth and on selected parameters of bacterial metabolism in digesta samples of piglets. AB - The effect of the probiotic strain Bacillus cereus var. toyoi on enterobacterial growth and selected metabolic parameters in digesta samples from piglets before and shortly after weaning was studied. Growth capacities of enterobacteria in digesta samples were significantly reduced, but no correlation was found between enterobacterial growth and concentrations of short-chain fatty acids and lactic acid, which are often influenced by other probiotic preparations. Tendencies for higher amounts of short-chain fatty acids were only recorded in digesta samples from the end of the jejunum, caecum and colon from piglets receiving probiotic supplemented feed. Lactic acid concentrations were reduced by the presence of B. cereus var. toyoi in feed in the first segment of the jejunum during the suckling period, while samples of the lower parts displayed an increased amount 4 days after weaning. Microbial activity of taurocholine deconjugating enzymes was significantly reduced in jejunum digesta from probiotic-fed piglets. There are strong indications that the early uptake of already modified microbial populations from faeces of the mother sow aids modification of intestinal microbial communities and their metabolic activities in piglets. PMID- 11906575 TI - Nutrition and diet in the clinical management of multiple sclerosis. AB - For many years, medical interest in the relationship between nutrition and multiple sclerosis (MS) has focused largely on aetiology and the influence of dietary fat on the rate and severity of disease. While the cause of MS remains unknown and the influence of dietary fat is unclear, recent studies on antioxidant intake and oxidative stress in MS are strengthening the rationale in support of a healthy eating regime following diagnosis. Dietary intake in MS and the influence of advanced disease on nutritional status are less well researched and documented. Both obesity and malnutrition may occur with detrimental consequences to functional abilities. Cognitive difficulties, dysphagia and the side-effects of drug treatment may further contribute to deterioration in nutritional status. This paper aims to provide a practical overview of dietary management in MS. It reviews the available evidence relating nutrition to MS and discusses dietary management, with particular emphasis on the identification and alleviation of factors affecting nutritional status. PMID- 11906576 TI - Improvement of symptoms in infant colic following reduction of lactose load with lactase. AB - Transient lactose intolerance has been identified as a possible causative factor in infant colic. A double-blind randomised placebo-controlled crossover study to investigate this has been undertaken in 53 babies with symptoms of colic. Pre incubation of the feed with lactase resulted in breath hydrogen levels and total crying time which were both at least 45% lower than figures with placebo treatment, in 26% of the full trial group (95% confidence interval 12.9% to 44.4%), and in 38% of compliers (95% confidence interval 18.8% to 59.4%). The remainder did not respond to the same extent. These findings suggest that infant colic may have a multiple aetiology, and that in a significant number of cases the immediate cause is transient lactose intolerance, in which cases pretreatment of feeds with lactase can result in considerable symptomatic benefits. PMID- 11906577 TI - Riboflavin deficiency in cystic fibrosis: three case reports. AB - Three cases of clinical riboflavin deficiency are reported in children aged 2-10 years attending a regional Cystic Fibrosis clinic. Riboflavin deficiency presented as angular stomatitis in all three patients. Patients were confirmed to be riboflavin deficient by assaying the activity of erythrocyte glutathione reductase. Patients were not on routine supplements of water-soluble vitamins before presentation and were treated with riboflavin supplements as part of a water-soluble vitamin complex. At presentation, one patient had poor nutritional status, but two patients were adequately nourished, receiving overnight Gastrostomy feeds. Data on these two patients indicate an adequate dietary intake of riboflavin, suggesting a mechanism for increased requirements, inadequate absorption or utilization. Additional deficiencies of thiamin, pyridoxine and iron were also observed. This paper reports the occurrence of a vitamin deficiency not previously reported in the cystic fibrosis population. PMID- 11906579 TI - The relative validity of a short Dutch questionnaire as a means to categorize adults and adolescents to total and saturated fat intake. AB - BACKGROUND: To assess the relative validity of a short food frequency questionnaire, the Fat list, to be used in (the evaluation of) nutrition education programmes. METHODS: Forty-five adults and 50 adolescents (12-18 years old) completed the Fat list at home, and subsequently kept diet records for seven subsequent days. RESULTS: Pearson correlations of about 0.7 for adults and 0.6 for adolescents were observed between fat scores derived from the Fat list and total and saturated fat intake in grams estimated by the 7-day diet records. Correlations varied among subpopulations based on sex, age, education, household size and responsibility for cooking and shopping. Lower correlations were especially found for female adolescents and older adolescents (16-18 years old). Pearson correlations between the Fat list and percentages energy from fat were low for both adults and adolescents. Gross misclassification, defined as disagreement between the two fat consumption assessments beyond an adjacent tertile, was less than 6% for all but the female adolescents. CONCLUSIONS: The Fat list can be used in adult and male adolescent populations to classify subjects in broad categories of total and saturated fat intake in grams and to assess differences in absolute and saturated fat intake between groups as a result of nutrition education programmes. PMID- 11906580 TI - Dietary intakes of elite female athletes in Greece. AB - BACKGROUND: Although there is a great interest in sports in Greece, there are very few data regarding dietary intakes and habits of Greek elite female athletes. The present study assesses the dietary intakes and the energy balance of elite female athletes of four different sports (volleyball, middle distance running, ballet dancing, and swimming) and a non-athletic control group. METHODS: Data were collected over two seasons, the training and the competitive, using 7 day weighed dietary records. Energy expenditure was calculated from 7-day activity records. Anthropometric measurements were also taken for all athletes. RESULTS: Athletes and controls had similar BMI values. Per cent body fat was lower for athletes compared with controls. Between sports, middle distance runners had the lowest per cent body fat. No significant differences were found between mean energy intake of athletes and controls. Mean energy intake was found lower than calculated energy expenditure, for all four teams. Macronutrient and micronutrient intakes of the athletes were not statistically different from those of the non-athletic control group. Mean micronutrient intakes were found above the recommended values with the exception of iron. Both athletes and controls had a high intake of vitamin C that is a characteristic of the population of the Mediterranean countries. CONCLUSIONS: Energy intakes varied between sports and between athletes of the same sport. Calculated energy expenditure was higher from the reported energy intake for most athletes. Athletes with the lowest energy intakes reported menstrual abnormalities. PMID- 11906578 TI - Family screening is effective in picking up undiagnosed Asian vitamin D deficient subjects. AB - Vitamin D deficiency has been described in the Asian migrants to the UK from the early 1960s. In spite of some suggestions that this problem is declining, we continue to see clinical cases of vitamin D deficiency with osteomalacia presenting to hospital. As the aetiology of this condition is associated with social, cultural and dietary factors, we screened associated family members of 18 index cases (three males 15 females, age range 12-73 years) who presented with clinical vitamin D deficiency to hospital. Of the 36 (21 females, 15 males) screened, 67% of these had evidence of vitamin D deficiency as judged by a 25(OH)VitD of < 5 microg L-1 (5-40). Some subjects also had hypocalcaemia (n=2), low PO4 (n=7), raised PTH (n=8) and raised alkaline phosphatase (n=11), indicating severe symptomatic, but unrecognized, vitamin D deficiency. Family screening seems an effective way of identifying Asian subjects with vitamin D deficiency who otherwise remain undiagnosed. A preventative policy with implementation is long overdue for this easily treatable condition. PMID- 11906583 TI - Diabetes--high time to assess dietetic effectiveness. PMID- 11906581 TI - Application of Hazard Analysis Critical Control Points system to enteral tube feeding in hospital. AB - An HACCP system was implemented for the quality assurance of preparation, storage and delivery of enteral feeds to patients in hospital. Routine methods of feed preparation, storage and delivery to patients were studied and a flow chart was initially made. After identifying hazards, an HACCP team was assembled, a flow chart was modified and critical control points were defined using a decision tree. Control measures for each step of the process and its monitoring and corrective measures to be applied were also defined. In addition, feed samples were analysed for microbiological quality and feed storage temperatures were also recorded, before and after the implementation of the HACCP system. When the control measures were applied and monitored, the hazard was reduced. Bacterial counts in feed were reduced from 105 cfu mL-1 to < 101 cfu mL-1. The results show that contamination of enteral feed may be reduced or eliminated if a systematic approach such as HACCP is applied effectively. PMID- 11906584 TI - The role of probiotics and prebiotics in the management of diarrhoea associated with enteral tube feeding. AB - INTRODUCTION: Diarrhoea is a common and serious complication of enteral tube feeding, and has a range of aetiologies. Manipulation of the colonic microflora may reduce the incidence of enteral tube feeding diarrhoea via suppression of enteropathogens and production of short-chain fatty acids. Probiotics and prebiotics are commonly used during enteral tube feeding to manipulate the colonic microflora; however, their efficacy is as yet uncertain. METHODS: English language studies investigating the pathogenesis of enteral tube feeding diarrhoea and the use of probiotics and prebiotics were identified by searching the electronic databases CINAHL, EMBASE and MEDLINE from 1980 to 2001. The bibliographies of articles obtained were searched manually. RESULTS: Only two prospective, randomized, double-blind, placebo-controlled trials have investigated the effect of a probiotic on enteral tube feeding diarrhoea; however, results are conflicting. No prospective, randomized, double-blind, placebo-controlled studies have specifically addressed the effect of a prebiotic on the incidence of enteral tube feeding diarrhoea. CONCLUSION: Theoretically, probiotics and prebiotics may be of benefit in prophylaxis against enteral tube feeding diarrhoea; however, there is currently insufficient evidence to support their routine use. Prospective, randomised, double-blind, placebo-controlled studies investigating their effect on diarrhoea are required. These observations are discussed with reference to the current literature. PMID- 11906585 TI - Framing of nutrition education messages in persuading consumers of the advantages of a healthy diet. AB - BACKGROUND: Educational dietary messages can stress either the positive consequences of performing a recommended dietary behaviour (positive frame) or the negative consequences of not performing a recommended dietary behaviour (negative frame). From studies on other health behaviours, there is evidence that positive frames have a stronger impact in encouraging preventive behaviours than negative frames. The main hypothesis of the present study was therefore that positively framed messages on eating a low-fat diet and eating enough fruit and vegetables (F & V) are more persuasive than negatively framed messages. METHODS: In a 2 (Frame: positive vs. negative) x 2 (Dietary behaviour: fat vs. F & V) design, 152 adult respondents randomly received one of four messages. Subsequently, they completed a questionnaire measuring cognitive attitude, affective attitude and intention to change the dietary behaviours. RESULTS: No significant differences in attitudes and intentions were found between the positive frame conditions and the negative frame conditions. CONCLUSIONS: Based on the current study no advice can be given yet to dietitians and other nutrition educators about whether they should emphasize the positive consequences of a dietary change or the negative consequences of not making the dietary change. PMID- 11906586 TI - Diabetes care in residential homes: staff training makes a difference. AB - AIM: To deliver an educational programme to care home staff in one residential home and assess its impact on staff knowledge and practice. METHODS: One home within Burnley, Pendle and Rossendale, East Lancashire, was randomly selected for education programme delivery and evaluation. An initial assessment questionnaire was used to develop the two, 2-h education sessions. Evaluation involved repeating the knowledge questionnaire 1 week and 12 months after the programme. A semi-structured interview with the officer in charge further investigated staff knowledge and care practice. RESULTS: There were 22 staff in the selected home; 12 participated in the programme. Pre- to post-course knowledge gain was significant (P < 0.001) and knowledge retention at 12 months was 92%. The officer in charge also reported at interview that quality of care had improved. CONCLUSIONS: Staff education increases knowledge and is associated with improved quality of care up to a year after the intervention. PMID- 11906587 TI - Cultural differences in assessing dietary intake and providing relevant dietary information to British African-Caribbean populations. AB - Diet can play a key role in the management of disorders such as obesity, diabetes and hypertension, conditions highly prevalent in the British African Caribbean population. In this paper, information not previously available is provided on the dietary habits and foods consumed by a British African-Caribbean population representative of the local community. Food frequency questionnaires were obtained from 255 randomly selected subjects in Manchester (78% of Jamaican origin), the nutrient intake results of which are available elsewhere. Here, suggestions are given to ensure that complete and valid dietary assessments (by diet history) are obtained, and the need for the approach to be somewhat different to that used in the White European population, highlighted with examples. Suggestions have also been listed for methods of dietary modification for obesity, diabetes and hypertension, taking into account differences in cultural understanding and food practices. People of Caribbean origin are not from just one territory: food habits and cultural context play an important role in every island, with clear differences between each which persist in first and later generations in Britain. In this paper, we attempt to integrate experience of learning from patients themselves during consultations and from participants in this study, with direct quantitative data on types of foods and their frequency in the local African-Caribbean diet. PMID- 11906588 TI - An investigation of the validity and reliability of a food intake questionnaire. AB - AIMS: To evaluate the reliability and criterion validity of a food intake questionnaire (FIQ) designed for use in schoolchildren. METHODS: Study of reliability: 98 young people aged 13-14 years attending two schools in deprived areas of Liverpool completed the FIQ on three separate occasions over a 3-month period. VALIDITY STUDY: Ninety-six young people (aged 11-13 years) completed the FIQ and 2 weeks later completed a 3-day food diary (with interview). RESULTS: The FIQ gave consistent response on separate occasions over the 3-month reliability study period. Levels of agreement were consistent between survey combinations. Analysis of variance showed no differences in mean score for food groups between surveys. Pearson correlations for mean scores estimated by separate FIQ ranged from 0.42 for fibre food group to 0.76 for negative marker food group; the majority of the correlations were above 0.5. The data suggested the FIQ should be able to detect a change of +/-10% in eating habits. The validity study provided modest but significant Pearson correlations between energy intake, fat intake as a percentage of energy intake and sugars intake derived from 3-day diaries, and mean scores for the fatty, sugary and negative marker food group assessed by the FIQ. CONCLUSIONS: The results from both studies provide an indication of the FIQ's reliability, and suggest it has criterion validity for fatty and sugary and negative marker foods. PMID- 11906589 TI - Meals and energy intake among elderly women--an analysis of qualitative and quantitative dietary assessment methods. AB - OBJECTIVE: The objective of this study was to analyse whether a qualitative method, in relation to traditional dietary assessment methods, was adequate to establish sufficient energy intake and energy content in separate meals in a population of elderly women. METHODS: One hundred and thirty-five elderly women, aged 63-88 years, living at home from three communities in mid-eastern Sweden participated in the present study. The quantitative methods used were a combination of repeated 24-h recall and a 3-day estimated food diary. The qualitative method used was the Food-Based Classification of Eating episodes model (FBCE). RESULTS: The mean intake of energy estimated by the 5-day registration was 6.8 +/- 1.9 MJ. The total number of eating events was 5.22 +/- 1.04 per day. On a group level, FBCE was useful to describe the diet among a group of elderly women; however, on an individual level, some complete meals were low or very low in energy, due to small portion sizes. CONCLUSION: The main conclusion was that a qualitative method, such as FBCE, must be supplemented with a dietary assessment method giving the energy intake to ensure that it is sufficient, especially when studying groups at risk for malnutrition. PMID- 11906590 TI - Dietitians and the internet: are dietitians embracing the new technology? AB - BACKGROUND: Internet use is increasing and in the future it will become a valuable tool for health professionals seeking and communicating health, diet and nutritional information. This study aimed to explore the use of the Internet by dietitians in the UK. METHODS: A cross-sectional study was carried out on a random sample of 200 dietitians selected from the British Dietetic Association database. A self-completion questionnaire was distributed to determine access to the Internet and the views of dietitians on the use of the Internet for the provision of health and nutrition information. RESULTS: Of the 156 respondents to the questionnaire, 96% were female and 4% were male. The results revealed that 66% of respondents had access to the Internet at work, with 39% using the Internet once a week or more. The main reported uses of the Internet were searching for health information, research and communication. Thirteen per cent of respondents reported seeing information obtained from the Internet by patients. Concerns about the use of the Internet for the provision of health information centred on creating unrealistic patient expectations. CONCLUSION: The research revealed that the Internet does have a role in augmenting current services. A substantial proportion of the dietitians studied were already using the Internet routinely in their work, with the real potential for its use beginning to be realized. PMID- 11906591 TI - Dietetic shortfalls that the National Service Framework for diabetes will need to address. PMID- 11906592 TI - Dietetic guidelines: diet in secondary prevention of cardiovascular disease (Journal of Human Nutrition and Dietetics, 14, 297-306). PMID- 11906594 TI - Managers, blame culture and highly ambitious policies in the British National Health Service (NHS). PMID- 11906595 TI - Work stress: an exploratory study of the practices and perceptions of female junior healthcare managers. AB - AIM: This exploratory study set out to investigate the perceptions and practices of junior healthcare managers with regard to stress at work. BACKGROUND: It has been suggested that cultural change is needed to accommodate a shift towards recognition of organizational responsibility for stress (Schulz et al. 1985). Logically, it can be argued that junior healthcare managers, as potential future senior managers, are best placed to facilitate this change. Junior healthcare managers' current thinking about stress had not yet been explored in depth. METHOD: A combination of critical incident diaries and semistructured interviews was conducted with six junior healthcare managers. The data were analysed and transcribed using a grounded theory approach. FINDINGS: The main themes to emerge were that junior healthcare managers were generally unaware of (a) potential work stressors and (b) the effect of work stressors on their own health and performance and that of their staff. CONCLUSIONS: The perceptions and practices of junior healthcare managers suggest that there is a culture of acceptance and expectation of work stress, combined with a lack of awareness to effectively and proactively manage it. PMID- 11906596 TI - Who would want to be a nurse? Violence in the workplace--a factor in recruitment and retention. AB - In a climate of a declining nursing workforce where violence and hostility is a part of the day-to-day lives of most nurses, it is timely to name violence as a major factor in the recruitment and retention of registered nurses in the health system. Workplace violence takes many forms such as aggression, harassment, bullying, intimidation and assault. Violent acts are perpetrated against nurses from various quarters including patients, relatives, other nurses and other professional groups. Research suggests that nurse managers are implicated in workplace violence and bullying. Furthermore, there may be a direct link between episodes of violence and aggression towards nurses and sick leave, burnout and poor recruitment and retention rates. This paper explores what is known about workplace violence as it affects nurses, and calls for managerial support and policy to act to improve work environments for all nurses. PMID- 11906597 TI - The essence of humour and its effects and functions: a qualitative study. AB - Humour is one of the innate abilities that an individual develops whilst growing up and which is affected by his/her experiences in life. The purpose of this study was to describe which categories can be included in the term 'humour' and to describe the effects and functions that humour has on people. The study was exploratory. The data were based on 20 interviews, nine of which were made with women and 11 with men who had no formal connection to health services or nursing. Ages ranged from 17 to 75 years and all the interviewees were from Sweden. The research question was: 'what does humour mean to you?'. The answers given were labelled as: laughter, happiness, unforeseen events/situations, real humour/art form, jokes, plays on words/puns, situation comedy and political satire. The categories were: possibilities/obstacles and weapon/protection. We conclude that the essence of humour can be categorized as possibilities/obstacles and weapon/protection. Humour has effects and functions on individuals. Empathy is a prerequisite for the use of humour in the context of health services and nursing. PMID- 11906598 TI - Using focus groups as a research method: a personal experience. AB - AIM: The personal experience and systematic process of using focus groups as a research method to assist change within an NHS Trust are described. BACKGROUND: Focus groups have recently emerged as a popular qualitative research method in health research and were used in a qualitative research study to explore, from an Enrolled Nurse perspective, what it was that prevented them coming forward for conversion to the First Level of the UKCC Nursing Register. METHOD: The author facilitated the five focus groups with Enrolled Nurse membership within the local organizational context. Data were recorded on audio-tapes and transcribed in preparation for analysis. RESULTS: The use of focus groups in health care research has key benefits for involving people in the research process and the subsequent changes that may occur as a result. The use of focus groups presented real challenges in the area of recruiting Enrolled Nurses to engage in such a process. CONCLUSIONS: Focus groups have a valuable role to play in both research and the involvement of people in organizational change and development. However, to persuade traditionally disempowered groups to engage in the process will be a challenge to the leaders of such projects. PMID- 11906599 TI - The influence of clinical supervision on ethical issues in home care of people with mental illness in Sweden. AB - AIM: To investigate in what ways clinical supervision can influence district nurses', psychiatric nurses' and mental health care workers' ethical decision making in home care of people with mental illnesses. BACKGROUND: Nursing staff frequently have to make difficult ethical decisions when caring for mentally disturbed patients in the home. METHODS: This study is a descriptive, correlational study. Data was collected by a cross-sectional survey that focused on psychiatric nurses, district nurses and mental health care workers (n = 660). RESULTS: Health care professionals, who received supervision as support in their clinical nursing work, perceived that they felt more secure in decision-making, felt safer in their relationship with the patient and had gained a deeper insight into the meaning of security for the patient as well as for the carer. Furthermore, they regarded taking over responsibility for the patient, when necessary, as their moral right and that care and treatment in the patient's home could mean that the patient's integrity was violated. CONCLUSIONS: The results emphasize the need for clinical supervision as support for nursing staff, as it leads to their acquiring a greater sense of self-esteem. There is also a need to clarify the professional role of nurses through integration of theoretical and clinical knowledge. The importance of supervision is illustrated by means of previous studies. PMID- 11906600 TI - The workshop as an effective method of dissemination: the importance of the needs of the individual. AB - The workshop is one of a number of strategies that can be used to disseminate information. This study set out to evaluate the effectiveness of the workshop as a method of disseminating information which had as its aim the introduction of new practices. A series of workshops, attended by G and H grade nurses, were carried out in a large teaching Hospital in England. The information which was disseminated concerned recruitment and retention activities for nurses which had been proposed in recent government documents, in particular in the Strategy for Nursing in England 'Making a Difference' by the UK Department of Health. Two sample groups of 25 nurses (workshop attendees and non-attendees) were interviewed. The interviews took place shortly after the workshops and were repeated 7 months later. The first set of interviews explored the nurses' intention to implement changes in their clinical areas and the second set of interviews identified actual change. The findings from the study suggests that the workshops were an effective means of producing changes in practice. However, the interviews also identified the value of personal engagement, especially in relation to meeting individual human needs, which were facilitated through the workshop. These emotions appeared to have an influence on the success of the nurses in implementing change and it is proposed that these outcomes should be considered when determining the benefits of a dissemination strategy. PMID- 11906601 TI - HSE research highlight that night workers need help to change their way of life. PMID- 11906602 TI - Rational design of alpha-helical antifreeze peptides. AB - The alanine-rich alpha-helical antifreeze protein from the winter flounder Pseudopleuronectes americanus adsorbs to specific planes of ice guided by an ice lattice match to threonine residues regularly spaced 16.6 A apart. We report here that by redesigning the winter flounder antifreeze peptide to incorporate a 27.1 A spacing between putative 'ice-binding' threonines, the deduced binding alignment of the helical molecule on the ice lattice is changed from the Miller indices directional vector [1102 ] to [2203 ]. Subsequent ice-binding characteristics are altered, including changes in adsorption specificity, decreases in thermal hysteresis activity and the formation of rotated hexagonal bipyramid ice crystal morphology. PMID- 11906603 TI - Peptides with indirect in vivo activity against an intracellular pathogen: selective lysis of infected macrophages. AB - A collection of natural peptides, simplified analogs of natural peptides, de novo amphipathic peptides and de novo amphipathic peptides composed of 50-80% alpha,alpha-dialkylated glycines (alpha,alpha-Dags) were synthesized on solid phase resin as the C-terminus amides using N-alpha-fluorenylmethyloxycarbonyl protection. The synthesis of the peptides rich in alpha,alpha-Dags used acid fluoride coupling methods. The peptides show antimicrobial activity against Escherichia coli and Staphylococcus aureus but no direct antimicrobial activity against Brucella abortus at 100 microm in vitro. However, in vivo treatment with several of these peptides results in significant reductions of B. abortus in chronically infected immune BALB/c mice relative to infected control animals. The chronically infected mice were susceptible to peptide toxicity at much lower peptide doses than control animals. The highest nonlethal dose for infected mice was only 25 microg for melittin, whereas 500 microg doses were nonlethal for many of the other peptides. Several of the alpha,alpha-Dag-rich peptides selectively destroy B. abortus-infected murine macrophages in vitro. Thus, these peptides apparently reduce the bacterial load in vivo by destroying a portion of the infected macrophages and exposing the sequestered bacteria to the immune response in the mice. PMID- 11906604 TI - Determination of stereochemistry stability coefficients of amino acid side-chains in an amphipathic alpha-helix. AB - We describe here a systematic study to determine the effect on secondary structure of d-amino acid substitutions in the nonpolar face of an amphipathic alpha-helical peptide. The helix-destabilizing ability of 19 d-amino acid residues in an amphipathic alpha-helical model peptide was evaluated by reversed phase HPLC and CD spectroscopy. l-Amino acid and d-amino acid residues show a wide range of helix-destabilizing effects relative to Gly, as evidenced in melting temperatures (DeltaTm) ranging from -8.5 degrees C to 30.5 degrees C for the l-amino acids and -9.5 degrees C to 9.0 degrees C for the d-amino acids. Helix stereochemistry stability coefficients defined as the difference in Tm values for the l- and d-amino acid substitutions [(DeltaTm' = TmL and TmD)] ranging from 1 degrees C to 34.5 degrees C. HPLC retention times [DeltatR(XL-XD)] also had values ranging from -0.52 to 7.31 min at pH 7.0. The helix-destabilizing ability of a specific d-amino acid is highly dependent on its side-chain, with no clear relationship to the helical propensity of its corresponding l-enantiomers. In both CD and reversed-phase HPLC studies, d-amino acids with beta-branched side chains destabilize alpha-helical structure to the greatest extent. A series of helix stability coefficients was subsequently determined, which should prove valuable both for protein structure-activity studies and de novo design of novel biologically active peptides. PMID- 11906605 TI - A possible mechanism for partitioning between homo- and heterodimerization of the yeast homeodomain proteins MATa1 and MATalpha2. AB - The yeast Saccharomyces cerevisiae has three cell types distinguished by the proteins encoded in their mating-type (MAT) loci: the a and alpha haploids, which express the DNA-binding proteins a1, and alpha1 and alpha2, respectively, and the a/alpha diploid which expresses both a1 and alpha2 proteins. In a/alpha cells, a1 alpha2 heterodimers repress haploid-specific genes and MATalpha1, whereas alpha2 homodimers repress a-specific genes, indicating dual regulatory functions for alpha2 in mating-type control. We previously demonstrated that the two leucine zipper-like coiled-coil motifs, called alpha2A and alpha2B, in the alpha2 N terminal domain are important to a1-alpha2 heterodimerization. A unique feature of alpha2B is the occurrence of three atypical amino acid residues at a positions within the hydrophobic core. We have conducted mutational analyses of alpha2B peptides and the full-length protein. Our data suggest that these residues may play a critical role in partitioning of the alpha2 protein between heterodimerization with a1 and homodimerization with itself. PMID- 11906606 TI - Structural analysis of fertilin(beta) cyclic peptide mimics that are ligands for alpha6beta1 integrin. AB - The NMR structural analysis of two fertilin(beta) mimics cyclo(EC2DC1)YNH2, 1, and cyclo(D2EC2D1C1)YNH2, 2 is described. Both of these mimics are moderate inhibitors of sperm-egg binding with IC50 values of 500 microm in a mouse in vitro fertilization assay. For peptide 1, the optimized conformations that best match the NMR data have a pseudo-type II' beta-turn with the linker and Glu at the i+1 and i+2 positions, respectively. The EC2D1C1 sequence is in a nonclassical (type IV) beta-turn. For peptide 2, the conformation that best matches the NMR data has two turns: a pseudo-type II' beta-turn in the D2EC2D1 sequence followed by a nonclassical beta-turn in the EC2D1C1 sequence. The Cbeta Cbeta distance between E and D1 in peptide 1 is 9.1 A, in peptide 2, it is 7.7 A. Thus, one possibility for the high IC50 values of these cyclic peptides is that the acidic residues are not constrained to a sufficiently tight turn, and thus much entropy must still be lost upon binding to the alpha6beta1 integrin. This explains why the cyclic peptides are the same as linear peptides at inhibiting sperm-egg binding. PMID- 11906607 TI - Topography of the neurotensin (NT)(8-9) binding site of human NT receptor-1 probed with NT(8-13) analogs. AB - A series of neurotensin (NT)(8-13) analogs featuring substitution of the Arg8 and/or Arg9 residues with non-natural cationic amino acids was synthesized and evaluated for binding to the human NT receptor-1 (hNTR-1). The modifications were designed to probe specific steric and electrostatic requirements in the N terminal cationic region of NT(8-13) for receptor binding as a general evaluation of the feasibility of incorporating minor structural changes into a peptide at a crucial polar receptor binding site. Many of the non-natural amino acids are more or less isosteric to Arg but more lipophilic as a result of addition of alkyl groups or through removal or replacement of NH character with methylene or methyl substituents, whereas others vary the distance between the cation and the alpha amino acid carbon. Substitution of Arg8 with N(G)-alkylated Arg derivatives or homolysine (Hlys) maintained the subnanomolar affinity of NT(8-13) to the hNTR-1. Position 8 incorporation of Hlys produced the most favorable primary amine side chain substitution to date. Moderate losses in affinity observed with position 9 substitutions were attributed to adverse steric effects. Doubly substituted [Hlys8, DAB9]NT(8-13), in which DAB is 2,4-diaminobutyric acid, was also prepared and tested as the shorter side-chain of DAB is known to be favored in position 9 of NT(8-13). This analog maintained 60% of NT(8-13) binding affinity making it the most favored des-guanidinium-containing analog known. These results demonstrate that adequate receptor binding affinity can be maintained over a structural range of Arg analogs, thus providing a range of peptides expected to exhibit altered pharmacokinetic properties. From the standpoint of the hNTR-1 cationic binding sites, these results help to map out the structural stringency inherent in the formation of a tight binding complex with NT(8-13) and related analogs. PMID- 11906608 TI - Double dimer peptide constructs are immunogenic and protective against Plasmodium falciparum in the experimental Aotus monkey model. AB - Multiple antigen peptide constructs (MAPs) have been used to obtain defined multimeric peptide molecules useful in the development of possible synthetic malaria vaccines. In this context, a method was developed, named double dimer constructs (DDCs), involving the direct synthesis of a dimeric peptide with a C terminal cysteine. A tetrameric molecule was then obtained by oxidation of sulfhydryl groups. Dimer synthesis was optimized using a Fmoc/tBu strategy, dimers were purified by HPLC, oxidized with DMSO and characterized by HPLC and MALDI-TOF-MS. The tetramers or DDCs obtained by this method were used as immunogens in the search for a possible malaria vaccine. It was found that they were immunogenic in the experimental Aotus monkey model, and were able to induce protective immunity when challenged experimentally with a highly infective Plasmodium falciparum malaria strain. PMID- 11906609 TI - Structure characterization of human serum proteins in solution and dry state. AB - The present report describes application of advanced analytical methods to establish correlation between changes in human serum proteins of patients with coronary atherosclerosis (protein metabolism) before and after moderate beer consumption. Intrinsic fluorescence, circular dichroism (CD), differential scanning calorimetry and hydrophobicity (So) were used to study human serum proteins. Globulin and albumin from human serum (HSG and HSA, respectively) were denatured with 8 m urea as the maximal concentration. The results obtained provided evidence of differences in their secondary and tertiary structures. The thermal denaturation of HSA and HSG expressed in temperature of denaturation (Td, degrees C), enthalpy (DeltaH, kcal/mol) and entropy (DeltaS kcal/mol K) showed qualitative changes in these protein fractions, which were characterized and compared with fluorescence and CD. Number of hydrogen bonds (n) ruptured during this process was calculated from these thermodynamic parameters and then used for determination of the degree of denaturation (%D). Unfolding of HSA and HSG fractions is a result of promoted interactions between exposed functional groups, which involve conformational changes of alpha-helix, beta-sheet and aperiodic structure. Here evidence is provided that the loosening of the human serum protein structure takes place primarily in various concentrations of urea before and after beer consumption (BC). Differences in the fluorescence behavior of the proteins are attributed to disruption of the structure of proteins by denaturants as well as by the change in their compactability as a result of ethanol consumption. In summary, thermal denaturation parameters, fluorescence, So and the content of secondary structure have shown that HSG is more stable fraction than HSA. PMID- 11906610 TI - Conformational investigation of alpha,beta-dehydropeptides. X. Molecular and crystal structure of Ac-DeltaAla-NMe2 compared with those of Ac-L-Ala-NMe2, Ac-DL Ala-NMe2 and other dimethylamides. AB - A series of three homologous dimethyldiamides Ac-DeltaAla-NMe2, Ac-L-Ala-NMe2 and Ac-DL-Ala-NMe2 has been synthesized and the structures of these amides determined from single-crystal X-ray diffraction data. To learn more about the conformational preferences of compounds studied, the fully relaxed (phi-psi) conformational energy maps in vacuo (AM1) of Ac-DeltaAla-NMe2 and Ac-L-Ala-NMe2 were obtained, and the calculated minima reoptimized with the DFT/B3LYP/6-31G** method. The crystal-state results have been compared with the literature data. Ac DeltaAla-NMe2 and other alpha,beta-dehydroamino acid dimethyldiamides, Ac DeltaXaa-NMe2 adopt the conservative conformation of the torsion angles phi, psi = approximately -45 degrees, approximately 130 degrees, which are located in the high-energy region (region H) of Ramachandran diagram. Ac-L-Ala-NMe2 and Ac-DL Ala-NMe2, as well as other saturated amino acid dimethylamides Ac-L/DL-Xaa-NMe2, present common peptide structures, and no conformational preferences are observed. Molecular packing of the amides analysed reveals two general hydrogen bonded motifs. Dehydro and DL-species are paired into centrosymmetric dimers, and L-compounds form catemers. However, Ac-DeltaAla-NMe2 and Ac-DL-Ala-NMe2 constitute exceptions: their molecules also link into catemers. PMID- 11906611 TI - Inhibition of human beta-tryptase by Bowman-Birk inhibitor derived peptides. AB - Four 11-residue peptides based on the Bowman-Birk inhibitor (BBI) structure were synthesized. These were tested for their ability to inhibit human beta-tryptase. Peptides with a basic residue at P1 inhibited tryptase even though the intact BBI protein is inactive. This result is interpreted in terms of the unique structural arrangement of active sites in tryptase which prevent access by large protein inhibitors. PMID- 11906612 TI - Transplantation for liver tumors: current status. AB - The question of liver transplantation for hepatobiliary malignancy has continued to generate controversial discussion. As shown by single-center studies and large databases, there is a clear indication for total hepatectomy and liver replacement under the premises of appropriate selection of suitable patients as well as of favorable type and stage of tumors. Future improvement of tumor-free patient survival can be expected from better understanding of tumor biology, including prevention and earlier detection of cancer, and effective multimodality treatment strategies. PMID- 11906613 TI - A novel role of alkaline phosphatase in protection from immunological liver injury in mice. AB - AIMS/BACKGROUND: Little is known about the role of alkaline phosphatase (AP) in liver diseases, except for its elevation in jaundice or cholestasis. Its substrate, endotoxin, is usually elevated in patients as well as animals with liver damage. This study aimed to provide evidence for its new role as protection against immunological liver damage. METHODS: Liver injury was induced in mice by delayed-type hypersensitivity to picryl chloride. AP activity was measured using a commercial kit. RESULTS: In acute liver injury, a significant decrease in AP activity in serum was observed but there was an increase in liver tissue. Single administration of cyclophosphamide before sensitization with picryl chloride exacerbated the liver injury, with more serious AP changes, while consecutive use after the sensitization alleviated the injury with a recovery from the changes. When liver injury proceeded for 1 week, both serum and liver showed decreased AP activity. Lipopolysaccharide facilitated alanine transaminase release from levamisole-pretreated but not non-treated hepatocytes from naive mice. However, the release was confirmed from liver slices of mice with liver injury proceeding for 1 week, even without levamisole pretreatment. CONCLUSION: The development of liver injury may lead to a dysfunction in AP synthesis and release. Levamisole may make normal hepatocytes, like the hepatocytes from liver-injured mice, highly sensitive to lipopolysaccharide through inhibiting AP synthesis. The findings obtained in this study suggest that AP may contribute to protection from injury by a mechanism involving neutralization of endotoxin. PMID- 11906614 TI - High efficiency gene transfer into cultured primary rat and human hepatic stellate cells using baculovirus vectors. AB - BACKGROUND/AIMS: Gene transfer into hepatic stellate cells (HSC) is inefficient when using plasmid-based transfection methods; viral-based systems are therefore being developed. A baculovirus system has recently been shown to be useful for expressing genes in mammalian cells. The aim of this study was to determine if baculovirus vectors can infect and express target genes in rat and human HSC and to assess potential cytotoxic and modulatory effects of infection. METHODS: A recombinant baculovirus vector (AcCALacZ) carrying the LacZ gene was used to infect HSC. beta-Galactosidase assays and electron microscopy were used to determine efficiency of infection and gene expression. Counting of trypan blue negative cells was used to assess cytotoxic/cytostatic effects of infection. Measurement of protein content of cells and alpha-smooth muscle actin expression were performed to assess the effects of baculovirus on cell function/phenotype. RESULTS: Baculovirus infection of activated HSC was highly efficient (> 90%) and provided long-term LacZ gene expression (15 days) in the absence of cytotoxic, cytostatic or modulatory effects. Infection of freshly isolated cells was also observed but at lower levels (20%). CONCLUSIONS: Baculovirus vectors can therefore be used to deliver target genes to cultured rat and human HSC with high efficiency and longevity in the absence of detrimental effects on cell function. PMID- 11906615 TI - Prognosis of a large cohort of patients with hepatocellular carcinoma in a single European centre. AB - BACKGROUND/AIM: Only a few follow up data are available for patients with hepatocellular carcinoma (HCC) in Europe and the USA. Therefore, we analysed all HCC patients admitted to our hospital between 1988 and 1999. METHODS: We documented aetiology, stage (HCC: Okuda and UICC classifications, liver cirrhosis: Child-Pugh score), and diagnostic and therapeutic measures of 281 consecutive HCC patients. Survival time was calculated as a function of staging and therapy. RESULTS: Cirrhosis was diagnosed in all patients. Seventy-two patients underwent liver resection, 28 liver transplantation, 31 transarterial chemoembolization and 14 percutaneous ethanol injection. One hundred and thirty six patients received no treatment. The Okuda and the Child-Pugh classification predicted a significant decrease of median survival time, whereas the UICC classification was less powerful. CONCLUSIONS: HCC occurred only in patients with liver cirrhosis. Survival time correlated with therapy (or no therapy) and with the Child-Pugh Score. In European patients the Okuda classification is superior to the UICC classification and should be compared to novel classification systems. PMID- 11906616 TI - Mannose-binding lectin and the prognosis of fulminant hepatic failure caused by HBV infection. AB - BACKGROUND/AIMS: The mannose-binding lectin (MBL) gene was reported to play an important role in determining the clinical outcome of persistent hepatitis B virus (HBV) infection. We investigated serum MBL concentrations and MBL gene mutations to determine whether they were related to the prognosis of patients with fulminant hepatic failure (FHF) caused by HBV infection. METHODS: We investigated serum MBL concentrations and MBL gene mutations in 43 HBV-infected Japanese patients with FHF and 260 HBsAg-negative healthy controls. Serum MBL concentrations were measured by an enzyme-linked immunosorbent assay, and mutations in the MBL gene were analysed by nested PCR and direct DNA sequencing. RESULTS: Only a mutation in codon 54 of the MBL gene was found. The frequency of this mutation in nonsurvivors (40%, 8/20) was higher than in survivors (13%, 3/23), and the difference was slightly significant (p = 0.043). The H allele frequency in survivors (70.5%, 31/44) was higher than in nonsurvivors (39.5%, 15/38) (p = 0.0048). Because of these factors the mean serum MBL concentration in survivors, 1.61 ,micro/ml (range 0.3-3.86), was significantly higher than in nonsurvivors, 0.79 microg/ml (range 0.04-1.51) (p < 0.0001). The likelihood ratio for nonsurvival was 0 for over 2.0 microg/ml, 0.67 for 1.0-2.0 microg/ml, and 2.24 for 0-1.0 microg/ml. CONCLUSIONS: The mutation in codon 54 of the MBL gene tended to be higher in nonsurvivors than in survivors. The H allele frequency (high producing allele in H/Y) in survivors was higher than that in nonsurvivors. High levels of serum MBL correlated with the survival of patients with FHF due to HBV infection. Serum MBL may be useful as a predictive factor for the survival of patients with FHF caused by HBV. PMID- 11906617 TI - Changes in distribution spaces and cell permeability caused by ATP in the rat liver. AB - AIMS/BACKGROUND: Cellular and extracellular volume changes caused by ATP were investigated in the liver as well as the possible formation of diffusion barriers, which could be responsible for some of its metabolic effects. METHODS: The experimental system was the bivascularly perfused rat liver. [(14)C]Sucrose and [(3)H]water were simultaneously injected into either the portal vein or the hepatic artery. Mean transit times, distribution spaces, variances and linear superimpositions were calculated. RESULTS: In the portal system, ATP reduced the transit time in the great vessels, had little or no effect on sinusoidal and cellular spaces, but impaired the flow-limited distribution of both [(14)C]sucrose and [(3)H]water. In the arterial bed ATP infused into either the portal vein or the hepatic artery produced vasodilation and increased the aqueous extra-sucrose space. These effects were inhibited by Nomega-nitro-L-arginine methyl ester infused into the hepatic artery. CONCLUSIONS: Sucrose and extra sucrose space changes caused in the arterial bed by portally infused ATP are most probably analogous to the transhepatic vasodilation effect already described for the rabbit liver. Impairment of flow-limited distribution of tracers in the sinusoidal bed indicates that ATP induces the formation of permeability barriers, which could be responsible for some of its metabolic effects. PMID- 11906618 TI - The mode of tumour progression in combined hepatocellular carcinoma and cholangiocarcinoma: an immunohistochemical analysis of E-cadherin, alpha-catenin and beta-catenin. AB - BACKGROUND/AIM: To investigate the mode of progression of combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CC). METHODS: An immunohistochemical study for E-cadherin (ECD) and alpha- and beta-catenins was performed on 29 cases of cHCC-CC. RESULTS: Reduced expression of ECD was significantly correlated with the tumour grade of the hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC) components, intrahepatic metastasis (IM) of HCC, IM of CC, and vascular invasion of CC (p < 0.05, respectively). There was a significant relationship between the reduced expression of beta-catenin and the tumour grade of HCC components (p < 0.05). Cases showing concurrent intrahepatic metastasis composed of HCC, CC, or both, numbered 6, 5, and 2, respectively. The expression patterns of ECD and beta catenin of IM were similar to those of primary lesion in most cases. On the other hand, expression of ECD and beta-catenin of IM of HCC component were preserved, even though those of the primary sites were reduced in two cases and one case, respectively. ECD and beta-catenin were significantly correlated with tumour differentiation and tumour progression. CONCLUSIONS: Preserved or recovered ECD and beta-catenin expression may be of beneficial effect for re-establishing the tissue architecture at the metastatic site. PMID- 11906619 TI - Fluorine-18 FDG imaging in hepatocellular carcinoma using positron coincidence detection and single photon emission computed tomography. AB - BACKGROUND/AIMS: We prospectively evaluated whether fluorine-18 deoxyglucose (FDG) positron coincidence detection (PCD) or FDG single-photon emission computed tomography (SPECT) provides additional benefits to our conventional preoperative evaluation of lesion detection in patients suspected to have hepatocellular carcinoma (HCC). METHODS: Thirteen consecutive patients with a suspected HCC underwent conventional preoperative evaluation with ultrasonography (US), triple phase helical computed tomography (CT), superparamagnetic iron oxides (SPIO) enhanced magnetic resonance imaging (MRI) and serum alpha-fetoprotein (AFP) level. All 13 patients had an FDG-PCD and SPECT. These results were evaluated to assess the value of FDG-PCD and SPECT in addition to US, SPIO-enhanced MRI and triple-phase helical CT. RESULTS: Ten of the 13 (77%) patients had at least one histologically confirmed HCC without extrahepatic abdominal spread. The tumors ranged in size from 1 to 8 cm and the serum AFP ranged from 3 to 30 000 microg/l. Of these 10 patients, two patients had an increased tumor F-FDG uptake (sensitivity of 20%); one patient with an AFP of 5 microg/l and a tumor size of maximum 4.5 cm and one patient with an AFP of 249 microg/l and a tumor size of maximum 2 cm. In three patients with a benign liver mass, FDG imaging with either PCD or SPECT was negative. There was no false positive finding. CONCLUSIONS: We found poor sensitivity of FDG-PCD and FDG-SPECT for the detection of HCC. There were no clear relations between AFP or tumor size and FDG uptake. Therefore, we conclude that FDG imaging with PCD or SPECT has no value in the preoperative work up for HCC in patients with cirrhosis. PMID- 11906620 TI - Expression of human telomerase reverse transcriptase in regenerative and precancerous lesions of cirrhotic livers. AB - BACKGROUND/AIMS: The catalytic subunit of human telomerase (hTERT) is known to be expressed in a variety of malignant tumours, including hepatocellular carcinoma (HCC). We studied hTERT expression in regenerative and precancerous lesions arising in cirrhosis. METHODS/RESULTS: As shown by in situ hybridisation, hTERT mRNA was absent in normal liver, but present in varying numbers of hepatocytes and HCC cells of diseased livers, as well as in biliary epithelial cells, lymphocytes, sinusoidal-lining cells and tumour endothelial cells. RT-PCR for two hTERT transcript regions demonstrated hTERT expression in 11 out of 15 cirrhotic liver samples, in 20 out of 21 large regenerative nodules/low-grade dysplastic nodules, in 5 out of 5 high-grade dysplastic nodules, and in 4 out of 4 HCCs. The beta-splice variant was identified in all hTERT-positive cases, while the corresponding full-length transcript was found only in 13 out of 29 positive large nodular lesions and in 4 out of 11 positive cirrhotic samples. The full length transcript was always found in the presence of the beta-splice variant, usually in low relative levels, and tended to correlate with telomerase activity in the samples, while the beta-splice variant did not. CONCLUSIONS: This study shows that hTERT re-expression takes place both in hepatic regeneration occurring in cirrhosis and in the early steps of hepatocarcinogenesis, and involves mainly the beta-splice variant of this molecule. Additional regulatory mechanisms may be required for the expression of the full-length hTERT transcript. PMID- 11906621 TI - Sclerosed hemangioma and sclerosing cavernous hemangioma of the liver: a comparative clinicopathologic and immunohistochemical study with emphasis on the role of mast cells in their histogenesis. AB - BACKGROUND/AIMS: Sclerosed hemangiomas of the liver are rare. To date, their histopathology, immunohistochemistry, and the role of mast cells (MC) in their histogenesis have not been systematically studied. PATIENTS/METHODS: Clinical, histopathologic and immunohistochemical features of 20 sclerosed hemangiomas were compared with those of 18 sclerosing cavernous hemangiomas. The number of MC was quantified and compared in all cases, using a tryptase immunostain. RESULTS: Compared to patients with sclerosed hemangiomas, those with sclerosing hemangiomas were younger (mean age, 63 versus 71 years); had larger tumors (mean 6 +/- 4.73 versus 3 +/- 2.2 cm); presented with a mass more frequently, and epigastric pain less frequently. Sclerosing hemangiomas, but not sclerosed hemangiomas, were more frequent in males than in females. Sclerosing hemangiomas occurred much more frequently in the right lobe than sclerosed hemangiomas. Sclerosing hemangiomas had less fibrosis, hyalinization, and elastic fibers than sclerosed hemangiomas (p = 0.00004). Numerous thick-walled blood vessels were a feature of sclerosed hemangiomas but not of sclerosing hemangiomas. Collagen IV, and laminin were more uniformly positive in sclerosing hemangiomas than in sclerosed hemangiomas. Increased immunoreactivity for smooth muscle actin was present in sclerosed hemangiomas more often than in sclerosing hemangiomas. FVIII R Ag, CD34, and CD31 were more diffusely positive in sclerosing hemangiomas than in sclerosed hemangiomas. In sclerosing hemangiomas, the mean number of tryptase positive MC per high power field (MC/HPF) varied from 8.25 +/- 6.23 in vascular areas to 1.6 +/- 4.01 in sclerotic areas. In comparison, the mean number of MC in sclerosed hemangiomas, was 4.3 +/- 5.01 in vascular areas, and 0.86 +/- 0.58 in sclerotic areas (p = 0.0095). The number of MC was significantly correlated with vascular proliferation and inversely related to the degree of fibrosis (p < 0.0001). CONCLUSIONS: This study demonstrates certain distinct clinical and histopathologic differences between sclerosing cavernous hemangiomas and sclerosed hemangiomas of the liver. We have established the presence of MC in those tumors, and suggest possible involvement of the MC in angiogenesis, and the regression process and development of fibrosis. PMID- 11906622 TI - Single nucleotide insertion in the 5'-untranslated region of hepatitis C virus with clearance of the viral RNA in a liver transplant recipient during acute hepatitis B virus superinfection. AB - Hepatitis C virus (HCV) infection is an important etiology in patients undergoing orthotopic liver transplantation (OLT) world-wide. Antiviral therapy-related clearance of HCV RNA may occur both in patients with chronic HCV infection and in transplanted patients for HCV-related liver cirrhosis, but the role of the 5' untranslated region (UTR) of HCV containing the internal ribosome entry site (IRES), which directs the translation of the viral open reading frame has not hitherto been evaluated. We studied the 5'-UTR in an HCV-infected recipient of a liver graft that showed spontaneous clearance of HCV RNA during an acute hepatitis B virus (HBV) superinfection. Sequencing of the 5'-UTR of HCV showed a nucleotide A insertion at position 193 of the IRES. PMID- 11906623 TI - Long-term asymptomatic biochemical cholestasis after fulminant or subfulminant liver failure is associated with extensive postnecrotic collapse with regeneration of the liver. AB - Fulminant or subfulminant liver failure usually leads to liver failure or to recovery. In rare instances, patients who recover exhibit prolonged asymptomatic biochemical cholestasis which coincides with the development into the parenchyma of large postnecrotic collapse with regeneration. This hitherto poorly recognized form of recovery may now be assessed by noninvasive techniques such as magnetic resonance imaging. We report the case of three patients who recovered from fulminant or subfulminant liver failure and in whom investigation of long-term biochemical cholestasis led to that unusual diagnosis. PMID- 11906624 TI - The detection of HBV antigens and HBx-transcripts in an Indian fibrolamellar carcinoma patient: a case study. AB - Fibrolamellar carcinoma (FLC) of the liver is a rare variant of hepatocellular carcinoma (HCC). Here we report the case of a 12-year-old Indian male with typical FLC with no apparent hepatitis B virus (HBV) infection and a non cirrhotic liver. The patient, though seronegative for HBsAg, showed expression of HBcAg in both the liver and tumour tissue. RT-PCR analysis revealed the presence of full-length HBx-transcripts in both liver/tumour tissue, along with truncated HBx-transcripts only in the tumour tissue. The lymphocytes in both peripheral and liver/tumour compartments showed a proliferative response to either/or HBcAg and HBxAg, which could be further augmented on addition of rIL-2. This is the first study to show not only the presence of HBcAg in the liver/tumour tissue but also prior exposure of the FLC patient's lymphocytes to HBV antigens. Also, the presence of the full-length and truncated HBx-transcripts in the tumour tissue, a proposed tumorigenic marker for hepatocarcinogenesis in chronic HBV patients, suggests an oncogenic role of HBV in this rare variant of HCC. PMID- 11906625 TI - Learning needs of Taiwanese nurses caring for patients with irritable bowel syndrome. AB - This study, building on work on irritable bowel syndrome conducted by Letson and Dancey (1996), explores Taiwanese nurses' knowledge, perceptions and beliefs about caring for patients with irritable bowel syndrome (IBS). A 46-item questionnaire was completed voluntarily by 120 registered nurses from a large tertiary acute care facility in Taiwan. The study used a descriptive research design and a questionnaire was developed that used a five-point Likert scale (strongly agree = 5, to strongly disagree = 1). The questionnaire consisted of six categories: demographic information; nurses' sources of IBS information; nurses' knowledge about IBS; nurses' perceptions about patients with IBS; nurses' beliefs about IBS; and learning requirements for nurses. Overall, the results indicted that Taiwanese nurses who participated in this study had little specific knowledge of IBS. The researchers developed a booklet containing information on the condition of IBS that may be used by the participants in this study to fill the knowledge gaps about this condition and provide useful information. PMID- 11906626 TI - Globalization of tertiary nursing education in post-Mao China: a preliminary qualitative assessment. AB - This article examines China's collaborative initiatives with Western countries to assess the impact of globalization on Chinese nursing education, especially at the post-secondary level, in the post-Mao era. Through the theoretical framework of mutuality, it evaluates the outcomes of globalization in two broad domains: pedagogy and system-institution-program building. In addition, case studies on two collaborative projects between Chinese nursing programs and Western institutions were conducted to further illustrate the principles of mutuality. This qualitative assessment is primarily based on a systematic review of published studies on the multifaceted dimensions of globalization in Chinese post secondary nursing education in both English and Chinese nursing literature since 1990. It is supplemented by unpublished documents and data obtained from a research trip to China in 2000. The study concludes that globalization has been, and will remain, one of the major forces underpinning Chinese nursing education (and the nursing profession in general), which is moving towards integration into the global nursing community. However, there is a significant imbalance in the knowledge transfer equation both in the national and international context. Great efforts need to be made to synthesize nursing knowledge in the East and West to achieve an integrative nursing science. PMID- 11906627 TI - Promoting screening and early detection of cancer in men. AB - Gender is a factor in the risk assessment for many diseases. It may also impact on the way in which men assess their personal health or illness status and take action to prevent illness or promote well-being. This paper is focused on three objectives: (i) to foster an understanding of gender differences in health promoting behaviors; (ii) to review three health issues affecting males for which dissemination of health education, increased personal awareness and early detection may be beneficial in the reduction of morbidity and mortality; and (iii) to offer suggestions for nurses and other health care professionals to promote positive patient-provider interactions within a health-care framework for action. PMID- 11906628 TI - Evaluating student learning: an Australian case study. AB - Determining the quality of student learning is an ongoing challenge to all educators. However, for educators and students in the health professions, evaluation of learning takes on a different dimension in terms of ensuring that graduates are competent, and thus safe, practitioners. This paper outlines the processes and methods by which student learning has been evaluated throughout a 22-year period at a large school of nursing in a Australian university. First, a historical overview of the major methods used demonstrates how relevant educational theories and sociopolitical forces and movements have influenced the whole curriculum including evaluation methods. Second, examples of current evaluation methods for undergraduate clinical and theoretical units are described. Reflections about past successes and future challenges conclude the paper. PMID- 11906629 TI - Challenges of recruiting a vulnerable population in a grounded theory study. AB - Recruitment is a crucial and fundamental part of research and one that poses various degrees of difficulty. This is particularly so when the area of research is one that is either highly sensitive, or that involves participants who are deemed to be particularly vulnerable. This article explores the inherent tensions in matters of participant recruitment among meeting the demands of institutional ethics committees, satisfying the concerns of clinicians in the field and the need to maintain methodological rigor. A postgraduate research student's experience of these tensions underpins the discussion. The article concludes with an outline of the student's strategies and resolution of these issues. PMID- 11906630 TI - Baccalaureate nursing education curricula in the People's Republic of China: status, issues and reforms. AB - This article examines baccalaureate nursing education curricula in the People's Republic of China. Three 5-year curricula and one 4-year curriculum were content analyzed and contrasted. Findings of this study suggested that: (i) the biomedical model dominated the three 5-year traditional curricula; (ii) political ideological content permeated all four curricula; and (iii) the reformed curriculum at Peking Union Medical College was a genuine effort toward building a new nursing curriculum model that intended to differentiate nursing education from medical education. In addition, major themes of recent curriculum reforms were highlighted. Recommendations were suggested to improve the outcomes of future curriculum endeavors. Explicit as well as implicit comparisons with an ideal-typical American generic baccalaureate nursing curriculum were made when appropriate. PMID- 11906631 TI - Alcohol consumption among 11-16 year olds: 'getting around' structural barriers? AB - This paper presents qualitative data from Irish children and adolescents on their experiences in relation to alcohol consumption. A sample of 78 participants (average age 11.5 years) was selected. A proportion of this initial sample were interviewed at intervals over a period of 3 years. The participants' consumption patterns were analyzed and four categories were generated: covert unsanctioned, overt unsanctioned, overt sanctioned, and peer unsanctioned. As the children got older, peer drinking became a stronger feature of the data; however, it mediated other patterns of behavior. Although the children displayed agency in circumventing adult rules relating to alcohol consumption, the participants were subjected to structural constraints by virtue of their status as children. Moreover, the agentic powers of the participants were procured through their social network rather than arising from an essentialist agency possessed by each individual child. The impact of childhood as a structural dimension weakened to some extent as the participants got older and had more freedom to circumvent adult-defined barriers to alcohol consumption. PMID- 11906633 TI - The role of interventional radiology in a pediatric liver transplant program. PMID- 11906634 TI - Neurodevelopment of infants with end-stage renal disease: is it improving? PMID- 11906636 TI - The emerging role of gene polymorphisms determining outcome after solid-organ transplantation. PMID- 11906635 TI - Hyporesponsiveness in pediatric recipients. PMID- 11906638 TI - Viral infections and pediatric liver transplantation. AB - Viral infections remain an important cause of morbidity and mortality following LTx in children. Increasing experience with both 'donor-associated' and 'community-acquired' viruses is leading to improved outcomes and the development of effective preventive strategies. The potential importance of emerging viral pathogens is an area of active research, which hopefully will also lead to effective treatments and potential preventive strategies for these newly recognized pathogens. PMID- 11906637 TI - Diagnosis and treatment of Wilson's disease. AB - Wilson's disease (WD) has moved on from being a recognized syndrome that was uniformly fatal to a curative disease for which the genetic basis has been discovered. Most pediatric patients present with hepatic manifestations, but some may have neurologic or psychiatric features. Clinical and biochemical screening, including liver biopsy for hepatic copper analysis, remain the standard for diagnosis, but haplotype analysis for siblings is now available and should be considered for family screening when possible. Lifelong medical therapy remains the mainstay of treatment, but treatment preferences are changing from penicillamine to alternative agents such as trientine and zinc. OLT remains lifesaving for those with fulminant WD and those in whom initial medical therapy fails. The future will probably see the application of rapid and accurate molecular diagnostic testing for this disorder and new therapeutic modalities such as hepatocyte transplantation, gene replacement therapy, and gene modification. PMID- 11906639 TI - Pediatric liver transplantation in metabolic disease: clinical decision making. AB - Proper utilization of liver transplantation in the management of pediatric metabolic diseases requires a comprehensive understanding of both metabolic disease and the risk and benefits of transplantation. This brief review focuses on issues that pertain to the treatment of tyrosinemia type I, bile acid biosynthesis disorders, primary hyperoxaluria, Crigler-Najjar Type I, and mitochondrial diseases. These entities are used as prototypes to illustrate many of the principles that are applied in a more general sense to the management of metabolic diseases. The natural history of these disorders are considered in the context of the risks of liver transplantation. Indications, contraindications, and both current and future alternatives to transplantation, are considered. PMID- 11906640 TI - Long-term outcome after liver transplantation in children. AB - Children (defined as under 18 yr of age) account for approximately 12.5% of all liver transplants in the United States. Even though the annual number of liver transplantation procedures remains relatively constant, the population of long term survivors of liver transplantation has grown. Presently, the population of long-term survivors of liver transplantation is 10-fold greater then the number of transplantations carried out each year. For long-term survivors of liver transplantation, the goal is to maintain graft function and wellness while decreasing the morbidity associated with long-term immunosuppression. The primary diagnosis leading to liver transplantation in children do not recur in the allograft. Consequently, many of the complications of liver transplantation, both early and long term, relate to the need for immunosuppression. Children may be at increased risk to develop significant end-organ damage as a result of increased serum lipid levels, elevated blood pressure, altered glucose metabolism, decreased renal function, cancer, and diminished bone accretion that occur as a result of immunosuppressive therapy or complications of therapy. As survival rates have increased, health care providers have begun to assess health-related quality of life. We will review our current knowledge of long-term outcome following pediatric liver transplantation in children. PMID- 11906641 TI - Prevention of parenteral nutrition-associated liver disease in children. AB - Liver injury is associated with parenteral nutrition therapy. Severity of injury varies from minimal and transient increases in liver-related blood tests to biliary cirrhosis and liver failure. Severe parenteral nutrition-related liver disease is usually confined to patients who have undergone massive intestinal resection. In these patients, early sepsis appears to cause initial liver injury, and recurring sepsis and inflammation, local or systemic, may result in its perpetuation and progression. Liver disease associated with parenteral nutrition is not necessarily related either to duration of parenteral nutrition or to delayed intestinal feeding. However, treatment includes enteral nutrition to promote enterohepatic circulation of bile acids and management of inflammation and sepsis, including control of intestinal bacterial overgrowth. Restriction of intravenous lipid emulsions may be important. The clinical picture of advanced liver failure related to short bowel syndrome differs from liver failure with an anatomically normal gastrointestinal tract. In the former, hyperbilirubinemia, hepatosplenomegaly, and functional hypersplenism dominate the clinical picture, and severe ascites and esophageal variceal hemorrhage are unusual. Early referral of these patients for intestinal and/or liver transplantation may provide the best chance for long-term survival. PMID- 11906642 TI - Living-donor liver transplantation in children. AB - With evolution of surgical technique, advances in immunosuppression, and better understanding of pre- and post-operative care, the 1-yr survival rate after liver transplantation in children has reached 85-90%. As a result, a greater number of patients have been listed for transplantation and waiting times have lengthened. Innovative techniques such as reduced-size, split, and living-donor liver transplantation are being applied more often to decrease long waiting times and reduce associated morbidity and mortality. In this review, living donor liver transplantation in pediatrics is described. Special issues, such as donor and recipient selection, surgical procedures in donors and recipients, and ethical issues, are discussed. PMID- 11906643 TI - Growth following solid-organ transplantation. AB - One of the ultimate goals of successful transplantation (Tx) in pediatric solid organ transplant recipients is the attainment of optimal final adult height. Except for kidney Tx there are limited data to address this issue. Remarkably similar factors impact on growth in pediatric kidney, liver, and heart recipients. Age is a primary factor, with younger recipients exhibiting the greatest immediate catch-up growth. Graft function is a significant contributory factor: a reduction in glomerular filtration rate (GFR) correlates with poor growth in kidney recipients, and the need for re-Tx is associated with impaired growth in liver recipients. The known adverse impact of corticosteroids on growth has led transplant physicians/surgeons to either modify the dose or attempt steroid withdrawal. In kidney and liver recipients this is associated with the development of acute rejection episodes. In infant heart transplant recipients the avoidance of maintenance corticosteroid immunosuppression is associated with normal growth velocity in the majority of recipients. With the marked improvement in patient and graft survival rates in pediatric solid-organ graft recipients, it is timely that the quality-of-life issues receive paramount attention. In children, normal growth following solid-organ Tx should be an achievable goal that results in normal final adult height. PMID- 11906644 TI - Neurodevelopmental outcome in high-risk patients after renal transplantation in early childhood. AB - Patient and graft survival rates of pediatric renal transplant recipients are currently excellent, but there are few reports regarding the long-term neurodevelopmental outcome after renal transplantation (Tx) in early childhood. Children with renal failure from infancy would be expected to have a less favorable developmental prognosis. We report the neurodevelopmental outcome in 33 school-age children transplanted between 1987 and 1995 when < 5 yr of age. We prospectively performed a neurological examination, magnetic resonance imaging (MRI) of the brain, electroencephalograms (EEGs), audiometry, and neuropsychological tests (NEPSY), and measured cognitive performance (WISC-R); we related these results to school performance and to retrospective risk factors prior to Tx. Twenty-six (79%) children attended normal school and 76% had normal motor performance. Six of the seven children attending a special school had brain infarcts on MRI. The EEG was abnormal in 11 (35%), and five (15%) received anti convulsive treatment after Tx. Sensorineural hearing loss was documented in six patients. The mean intelligence quotient (IQ) was 87, and 6-24% showed impairment in neuropsychological tests. The children attending a special school had been more premature, but had not had a greater number of pre- or neonatal complications. They had experienced a greater number of hypertensive crises (p = 0.002) and seizures (p = 0.03), mainly during dialysis, but the number of septic infections and the mean serum aluminum levels were not significantly greater than in the children with normal school performance. In these previously lethal diseases, the overall neurodevelopmental outcome is reassuring. However, it is of crucial importance to further minimize the risk factors prior to Tx. PMID- 11906645 TI - High-dose peripheral blood stem cell infusion: a strategy to induce donor specific hyporesponsiveness to allografts in pediatric renal transplant recipients. AB - We designed and implemented a clinical trial to achieve zero-rejection status in pediatric renal allograft recipients, using granulocyte-macrophage colony stimulating factor (GM-CSF)-stimulated peripheral blood stem cell (PBSC) infusion. We studied 44 consecutive patients: 24 volunteers in a treated group (Tn) and 20 in a control group (Cn). Both groups were comparable with respect to clinical and laboratory parameters. The Tn group had 70.8% one haplo-match donors and the Cn group had 80% one haplo-match donors. Patients in the Tn group received cyclosporin A (CsA) and 0.4 mg/kg body weight prednisolone as immunosuppressants; azathioprine was added for patients of the Cn group, who received 1 mg/kg body weight prednisolone together with CsA. Living-related donors (LRD) of patients in the Tn group received GM-CSF 450 microg on four consecutive days followed by leucopheresis and immediate transfusion of unmodified PBSC into the recipient. This procedure was repeated once/twice, with one portal and one/two systemic infusions. Our aim was to maximize the dose of PBSC. The total average dose was 22 x 10(8) cells/kg body weight. Lymphocyte cross-match (LCM) was performed before GM-CSF injection and after the last PBSC infusion. Follow-up over an 18-month period revealed 100% graft survival with sustained low serum creatinine (SCr) values in patients of the Tn group as compared with 80% graft survival in patients of the control group who had marginally higher SCr levels. Absence of graft vs. host disease (GvHD), acute rejection episodes, and low incidence of cytomegalovirus (CMV) disease were the principal benefits of this protocol. PMID- 11906647 TI - Living-related liver transplantation for biliary atresia associated with polysplenia syndrome. AB - This report describes a 1-yr-old boy with biliary atresia (BA) and polysplenia syndrome (PS) who underwent successful living-related liver transplantation (LTx). At the time of initial hepatic portoenterostomy, he was noticed to have a preduodenal portal vein (PV), non-rotation of the intestine, and polysplenia. Because he did not achieve good bile excretion, he underwent a living-related LTx (using a left lateral segment from his mother) at the age of 14 months. Evaluation of the vascular anatomy was made by angiography, magnetic resonance imaging (MRI), computerized tomography (CT), and Doppler ultrasound. The PV was stenotic from the confluence of the superior mesenteric vein (SMV) and splenic vein (SpV) to the hepatic hilum. The retrohepatic inferior vena cava (IVC) was deficient cranially to the renal vein and was connected to the azygous vein. The supra-hepatic IVC was detected below the diaphragm and was connected to three hepatic veins. The common hepatic artery (HA) originated from the superior mesenteric artery. At LTx, the PV was dissected to the level of confluence of the SMV and the SpV, from which the venous graft was interposed using the donor's ovarian vein. Three hepatic veins were plastied into one orifice, which was anastomosed to the graft's hepatic vein under the diaphragm. The graft vascularity and function has been good for 1 yr after LTx. In the present case, sufficient pre-LTx evaluation of vascular anomalies seemed to help performance of the successful LTx. PMID- 11906646 TI - Association of cytokine single nucleotide polymorphisms with B7 costimulatory molecules in kidney allograft recipients. AB - African-American race is associated with an increased risk of allograft loss, suggesting that African-American patients may form an immunologically higher risk group. Previously, we demonstrated that immune cell costimulatory molecule expression is significantly higher in African-Americans than in Caucasians. Polymorphic variations in the genes for cytokines have been associated with a number of immunological conditions, and with transplant rejection. This study was performed to determine the distribution, in African-American and Caucasian renal transplant recipients, of single nucleotide polymorphisms (SNPs) in the following cytokine genes: tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma), interleukin (IL)-6, IL-10, and transforming growth factor-beta (TGF beta). Cytokine production from blood cells was determined, and cell-surface B7 (CD80, CD86) expression was measured. There was a significant link between IL-10 genotype and acute rejection episodes, but only in African-American patients (p < 0.01). Also, African-American patients had a significantly higher probability of having the IL-6 G-allele (p < 0.0001), which is associated with a high production of IL-6 protein. Incubation of blood cells with IL-6 resulted in increased expression of surface CD80 and CD86, while IL-10 decreased CD80 expression. This study demonstrated a clear correlation of the IL-6 G-allele with increased cellular CD80 expression and the IL-10 G-allele with decreased CD80 expression. These data raise the possibility that specific genotypes are associated with local cytokine regulation of cell-surface costimulatory molecule expression. African-American patients may have a genetically determined, quantitatively different immune response than Caucasian patients, contributing to adverse transplant outcomes. PMID- 11906648 TI - Post-biopsy renal arteriovenous fistula. AB - A 6-yr-old boy developed progressively severe hypertension, which was unresponsive to medications, 1 week after percutaneous biopsy of his renal transplant. Renal angiogram revealed an arteriovenous fistula (AVF) in the lower pole at the site of the biopsy. Case findings and resolution after embolization are described, and the current literature is briefly reviewed. PMID- 11906649 TI - Sebaceous adenitis in the Akita: clinical observations, histopathology and heredity. AB - Ninety-seven pure-bred Akitas were examined clinically and histologically for sebaceous adenitis. The diagnosis was established histologically in 23 Akitas by demonstrating an inflammatory reaction targeted against the sebaceous glands or a reduction in the number of glands. The clinical course of sebaceous adenitis in the Akita was similar to that seen in other breeds. The first skin lesions occurred mainly on the dorsal midline and ears. Compared with the Poodle, the age at first onset of the disease was more variable and the hair loss affected mainly the undercoat. The progression of sebaceous gland destruction varied between dogs and was not seen in all cases. Because bud-like sebaceous gland proliferation could be identified, it seems that regeneration of the sebaceous glands may occur. An autosomal recessive inheritance appears to be possible. Apart from a genetic background, immune-mediated factors possibly influence the onset and course of sebaceous adenitis. PMID- 11906650 TI - A retrospective study of canine and feline cutaneous vasculitis. AB - Twenty-one cases of cutaneous vasculitis in small animals (dogs and cats) were reviewed, and cases were divided by clinical signs into five groups. An attempt was made to correlate clinical types of vasculitis with histological inflammatory patterns, response to therapeutic drugs and prognosis. Greater than 50% of the cases were idiopathic, whereas five were induced by rabies vaccine, two were associated with hypersensitivity to beef, one was associated with lymphosarcoma and two were associated with the administration of oral drugs (ivermectin and itraconazole). Only the cases of rabies vaccine-induced vasculitis in dogs had a consistent histological inflammatory pattern (mononuclear/nonleukocytoclastic) and were responsive to combination therapy with prednisone and pentoxifylline, or to prednisone alone. Most cases with neutrophilic or neutrophilic/eosinophilic inflammatory patterns histologically did not respond to pentoxifylline, but responded to sulfone/sulfonamide drugs, prednisone, or a combination of the two. PMID- 11906651 TI - A randomized, double-blind, placebo-controlled trial to investigate the efficacy and safety of a Chinese herbal product (P07P) for the treatment of canine atopic dermatitis. AB - A randomized, double-blind, placebo-controlled trial of P07P, a product derived from a traditional Chinese herbal remedy, was undertaken in 50 dogs with atopic dermatitis. Owners recorded a daily itch score for 4-14 days before treatment and during treatment. Packets of powder containing P07P or placebo were added to the food once daily for 8 weeks. Dogs were assessed for erythema, surface damage, overall coat condition and seborrhoea by the same investigator, as well as for pruritus and general demeanour, at 0 (visit 2), 28 (visit 3) and 56 (visit 4) days of treatment or at withdrawal. Investigator and owner assessments of response were recorded after 28 and 56 days of treatment or at withdrawal. The predefined primary outcome measure was the owners' assessment of response at the end of treatment. Nine of the 24 dogs (37.5%) in the P07P group but only 3 of the 23 dogs (13%) in the placebo group were considered to have improved, but this difference was not statistically significant (P = 0.09). There was a significantly higher withdrawal rate due to worsening of condition in the placebo group (P = 0.04). Mean daily itch score in the second 28-day period of the study was significantly higher than baseline in the placebo group (P = 0.01) but not in the P07P group (P = 0.30). Pruritus scores showed a significant deterioration from baseline at the final visit in the placebo group (P = 0.01) but not in the P07P group (P = 1.00). There was a significant difference between the groups in change from baseline in erythema score at visit 3 (P = 0.05). There were no significant differences (P > 0.05) in surface damage, seborrhoea, overall coat condition and general demeanour scores within or between the groups throughout the study. The product was well tolerated with no severe or serious adverse events recorded. P07P may be beneficial as a novel nonsteroidal therapy for the management of dogs with atopic dermatitis. PMID- 11906652 TI - Scanning electron microscopy description of a new species of Demodex canis spp. AB - Between 1997 and 1999, the prevalence of Demodex canis mites was determined in 150 dogs. In two dogs, we found two different species of mites; Demodex canis and another, unidentified, Demodex mite. The unidentified Demodex mite species had several different morphological features. First, it had a short opisthosoma and an obtuse end. In addition, the fourth coxisternal plate was rectangular and there was a band-like segmental plate between the fourth coxisternal plate and opisthosoma. Although all of the morphology and the development of male mites could not be investigated in this study, the location of the opisthosoma and the genital pore clearly differed from Demodex canis, suggesting that this unidentified mite is a new species. PMID- 11906653 TI - Degenerative mucinotic mural folliculitis in cats. AB - A novel form of mural folliculitis is described in seven cats. Clinically, all cats exhibited generalized alopecia with scaling or crusting that was more pronounced over the head, neck, and shoulders. The face and muzzle of all cats was unusually thickened. Six of seven cats were progressively lethargic but did not demonstrate any other consistent systemic abnormalities. Histologically, there was severe mixed inflammation of the wall of the follicular isthmus in all cats, accompanied by some follicular destruction in five cats. Sebaceous glands were not affected. All cats had variable, but often striking, follicular mucin deposition, as well as epidermal hyperkeratosis and crusting. The cause of the severe mural folliculitis was not identified, and all cats responded poorly to immunomodulating therapy. Follicular mucinosis may be a nonspecific finding, likely reflective of the follicular lymphocytic milieu, and does not always herald follicular lymphoma. PMID- 11906654 TI - Epidermal dysplasia and Malassezia infection in two West Highland White Terrier siblings: an inherited skin disorder or reaction to severe Malassezia infection? AB - Two 9-month-old West Highland White Terrier siblings were referred to our clinic with pruritus, alopecia and lichenification. Cytological examination of Scotch tape strippings revealed Malassezia organisms and cocci. Skin biopsy specimens showed epidermal dysplasia. Treatment included bathing with a 2% miconazole/chlorhexidine-containing shampoo, orally administered ketoconazole (5 mg kg-1, every 12 h) and cloxacillin (25 mg kg-1 every 8 h). Six weeks later, the dermal infection had resolved and there was hair regrowth. However, the dogs were still moderately pruritic. Intradermal allergy testing was positive for house dust mites, storage mites and Malassezia. Immunotherapy was initiated, and treatment with ketoconazole and cloxacillin was stopped. Skin biopsies, which were performed in both dogs 4 months after the first presentation, revealed mild superficial perivascular dermatitis. The remaining mild facial pruritus was easily controlled with topical treatment. These two cases indicate that epidermal dysplasia might be an inflammatory or hypersensitivity reaction to the Malassezia infection or a result of excessive self-trauma, rather than a congenital keratinization disorder. PMID- 11906655 TI - Spongiotic vesicular dermatitis as a cutaneous reaction pattern in seven horses. AB - Over a 6-year period seven adult horses of different breeds and genders developed multifocal, exudative, oozing dermatitis characterized histologically by epidermal spongiotic vesicles and perivascular eosinophilic, neutrophilic and mixed mononuclear inflammation. Three horses were pruritic. Systemic disease was not noted. Two horses had a history of recurrent urticaria (hives) and one horse had nodules or welt-type lesions that progressed to exudative, oozing lesions. Interepithelial immunoglobulin (Ig)G was detected by avidin-biotin complex peroxidase staining, but the pattern of staining was more consistent with epithelial oedema than specific IgG deposition associated with pemphigus. The exudative oozing lesions developed under circumstances suggesting that dermal oedema progressed to intracellular and intercellular epidermal oedema, which in turn progressed to the spongiotic vesicular epidermal lesions. PMID- 11906656 TI - A case of feline phaeohyphomycosis due to Fonsecaea pedrosoi. AB - The first report of a case of feline phaeohyphomycosis due to Fonsecaea pedrosoi is presented. Fonsecaea pedrosoi is an aetiologic agent of both human phaeohyphomycosis and chromoblastomycosis. In our cat, the lesion was confined to the skin and appeared as a firm swelling on the bridge of the nose. Diagnosis was based on histological examination of a cutaneous biopsy and fungal culture of a tissue sample on Sabouraud's dextrose agar. Further diagnostic tests failed to reveal an underlying immunosuppression. Two treatment cycles with itraconazole, at the oral dose of 5 mg kg-1 given twice daily, induced complete clinical remission, but relapses occurred. PMID- 11906657 TI - Current DNA-based tests for hereditary eye disease. PMID- 11906658 TI - Periocular neurofibrosarcoma in a horse. AB - A periocular neurofibrosarcoma was debulked and treated with intralesional cisplatin in a 5-year-old Thoroughbred mare. The horse presented with a 1-year history of a large slowly progressing subcutaneous mass over the right supraorbital process. The mass was surgically debulked, and intralesional cisplatin (1.0 mg/cm3) was injected in 3 doses at 2 weeks, 5 weeks, and 8 weeks postoperatively. No recurrence was noted over a 15-month follow-up period. Histopathology of the mass indicated neurofibrosarcoma. PMID- 11906659 TI - Functional MRI as a tool to assess vision in dogs: the optimal anesthetic. AB - Functional magnetic resonance imaging (fMRI) is a recent advance in neuroimaging that provides a picture of brain activity with excellent spatial resolution. Current methods used to evaluate canine vision are poorly standardized and vulnerable to bias. Functional MRI may represent a valuable method of testing vision in dogs if the impacts of anesthesia on fMRI are understood. Six dogs were scanned during visual stimulation, each under three different anesthetic protocols (isoflurane, propofol, fentanyl/midazolam) to address the questions: (1) Can visually evoked fMR signals be reliably recorded in anesthetized dogs? and (2) Which anesthetic agent permits the least suppression of visually induced fMR signal in dogs? This study confirms that visual stimuli reliably elicit neural activity and fMR signal change in anesthetized dogs. No significant differences in images acquired under the three anesthetics were found, and there was no significant relationship between anesthetic dose and brain activity, within the range of doses used in this study. Images obtained during isoflurane anesthesia were more consistent between dogs than those obtained with the other two agents. This reduced variation may reflect the fact that inhalant anesthesia is more easily controlled than intravenous anesthesia under conditions associated with high field fMRI. PMID- 11906660 TI - A case of orbital hemangiopericytoma in a dog. AB - A 7-and-a-half-year-old-dog was presented with progressive unilateral exophthalmos. Computed tomography imaging revealed an orbital mass that was surgically excised by lateral orbitotomy to preserve vision. The tumor was diagnosed histologically as a hemangiopericytoma. Twelve months postoperatively there were no signs of a local recurrence. This is the first case report of a hemangiopericytoma involving the orbit of a dog. PMID- 11906661 TI - Tolerance of the rabbit cornea to an n-butyl-ester cyanoacrylate adhesive (Vetbond). AB - OBJECTIVE: An n-butyl-ester cyanoacrylate adhesive available for veterinary surgery (Vetbond, 3M) was tested in rabbits for corneal irritation. PROCEDURES: Two experimental procedures were used on 24 rabbits: injection of the adhesive into an intralamellar corneal pocket (n = 10) and application of the glue to a mid-stromal corneal defect (n = 14). In both experiments the eyes were examined for 20 days for evidence of corneal irritation and tolerance. At the end of each experiment, histopathologic studies were performed on all corneas. RESULTS: The corneal reaction to the intrastromally injected cyanoacrylate was characterized clinically by slight edema and vascularization localized to the vicinity of the adhesive. A moderate foreign body-type reaction was found histologically. Following application of the adhesive to a central stromal defect, the treated corneas remained totally clear and histopathologic examination showed that the healing process was not altered compared to the controls. The mean retention time of the glue patch was 14 days. CONCLUSIONS: Intrastromal injection and surface application to a corneal defect of n-butyl-ester cyanoacrylate to a corneal defect induced only a mild inflammatory response and did not interfere with the reparative process. These findings suggest that this surgical adhesive would be acceptable for treating corneal ulcerations in animals. PMID- 11906662 TI - Postnatal development of corneal curvature and thickness in the cat. AB - OBJECTIVE: To evaluate the postnatal development of central corneal curvature and thickness in the domestic cat. Animals studied Six Domestic Short-haired (DSH) kittens starting at 9 weeks of age and 6 adult cats. PROCEDURES: Kittens were evaluated biweekly to monthly for a 12-month period, starting at age 9 weeks. Corneal development was monitored by hand-held keratometry and ultrasound biomicroscopy. Standard regression analysis using a nonlinear least squares method was used to generate a formula that would predict corneal curvature as a function of age. RESULTS: Mean keratometry (K) values for the 9-week-old cats were 54.51 (+/-1.02) diopters (D) and these values steeply declined over the next 3 months to 44.95 (+/-0.90) D. Thereafter, K-values gradually decreased to reach a plateau by 12-15 months of age of 39.90 (+/-0.42) D. Because K-values still appeared to be slightly diminishing at this point, six other > 2-year-old cats were evaluated by keratometry and were found to have K-values of 38.99 (+/-0.81). Two to four diopters of astigmatism was common in young kittens whereas adult cats had a low mean degree of astigmatism (< 1 D). A formula that predicted keratometry values in diopters (K) as a function of age in weeks (w) was established as follows: K = 39.83 + 26.87 exp(-0.074 w). The central cornea increased in thickness primarily during the first 4 months of life with 9 week old kittens having values of 0.379 (+/-0.012) mm; 16-week-old kittens, 0.548 (+/ 0.021) mm and 67 week-old cats, 0.567 (+/-0.012) mm. CONCLUSIONS: The maturation process of the feline cornea proceeds over the first 1-2 years of life to attain an adult status that is characterized by a roughly spherical state of approximately 39 D corneal curvature, substantially flatter than the human cornea, and a central thickness similar to the human cornea. Research studies of the refractive or optical properties of the cornea in which cats are used as experimental animals should be conducted on animals greater than 18 months of age. PMID- 11906663 TI - The relationship of cataract maturity to intraocular pressure in dogs. AB - OBJECTIVE: To determine the distribution of intraocular pressure, as measured by applanation tonometry, in dogs with cataracts, and compare these tonometric results to the different stages of cataract formation (incipient, immature, mature, and hypermature). Animals studied Retrospection study of canine clinical patients (86 dogs). PROCEDURES: All records of dogs presented from 1991 to 1996 to the university veterinary medical teaching hospital for diagnosis of cataracts and evaluation for cataract surgery were reviewed. The tonometric measurements from the initial ophthalmic examination were selected in cataractous and nonglaucomatous eyes either receiving no topical or no systemic medications. The stage of cataracts was based on the degree of opacification, tapetal reflection, clinical vision, and visibility of the ocular fundus by indirect ophthalmoscopy. The distribution of tonometric results were grouped by the cataract maturity, and compared by anova and Tukey's general linear tests. RESULTS: Intraocular pressure with incipient cataracts ranged from 9 to 17 mmHg (mean 12.7 +/- 1.2 mmHg). Intraocular pressure with immature cataracts ranged from 3 to 27 mmHg (mean 13.6 +/- 0.6 mmHg). For the mature cataracts, IOP ranged from 5 to 22 mmHg (mean 11.9 +/- 0.7 mmHg). For the hypermature cataract group, IOP ranged from 4 to 23 mmHg (mean 10.8 +/- 0.6 mmHg). Comparison of the tonometric results among the different stages of cataract formation indicated a significant difference (P = 0.0086) between only the immature and hypermature groups. CONCLUSIONS: Intraocular pressure in lens-induced uveitis (LIU) is lowered but the relationship to the stage of cataract maturity is less clear. Significant tonometric differences were present between the immature and hypermature cataract groups, but these differences are too small to be clinically useful. Decreased intraocular pressure of dogs with all stages of cataract formation suggests concurrent LIU during all stages of cataract formation, especially with the mature and hypermature stages. The average tonometric measurements in dogs with these cataracts were about two standard deviations below the mean IOP reported in normal dogs. PMID- 11906664 TI - The electroretinographic phenotype of dogs with Golden Retriever muscular dystrophy. AB - OBJECTIVE: To characterize the flash electroretinogram (ERG) in the Golden Retriever muscular dystrophy (GRMD) dog and to compare the results with those from a control group of Golden Retrievers. To investigate whether similar abnormalities of the ERG as those found in a majority of human patients with Duchenne muscular dystrophy (DMD) are also observed in the GRMD dog, the canine model for DMD. Animals Five GRMD dogs and five age-matched clinically normal Golden Retrievers. PROCEDURE: An ophthalmic examination was carried out prior to performing electroretinography under general anesthesia. Rod, combined rod-cone and oscillatory potentials responses were recorded after dark adaptation. Responses to 30-Hz-flicker were recorded after light adaptation. The ERG responses of the GRMD dogs were compared with those of the control dogs by use of a Wilcoxon signed rank test. RESULTS: GRMD dogs had significantly reduced a and b wave amplitudes after dim white flash stimuli (rod response) and reduced a-wave amplitude after bright white flash stimuli (rod-cone response). CONCLUSION AND CLINICAL RELEVANCE: The ERG abnormalities observed in the GRMD dog suggest a dysfunction in the rod signaling pathway. These ERG alterations are different from those observed in human patients with DMD. PMID- 11906665 TI - Effect of different dose schedules of latanoprost on intraocular pressure and pupil size in the glaucomatous Beagle. AB - OBJECTIVE: To evaluate the changes in intraocular pressure and pupil size in glaucomatous dogs after instillation of 0.005% latanoprost (Xalatan, Pharmacia and Upjohn, Kalamazoo, MI, USA) once in the morning, or once in the evening, or twice daily in five-day multiple-dose studies. Animals studied Eight Beagles with the moderate stage of inherited primary open-angle glaucoma. PROCEDURES: Applanation tonometry (IOP) and pupil size (PS) measurements were obtained at 8 am, 10 am, 12 noon, 2 pm, and 4 pm in eight glaucoma dogs. Methylcellulose (0.5% as placebo) was instilled in the control eye, and 0.005% latanoprost was instilled in the opposite drug eye. Control and drug eyes were selected using a random table. For these three studies, 0.5% methylcellulose and 0.005% latanoprost were instilled the second through the fifth days with instillations in the morning (8.30 am), or evening (8 pm), or twice daily (8.30 am and 8 pm). Statistical comparisons between drug groups included control, placebo, and treated (0.005% latanoprost) eyes for three multiple-dose studies. RESULTS: In the 8-am latanoprost study, the mean +/- SEM diurnal declines in IOP for the placebo and drug eyes for the first day were 6.5 +/- 3.6 mmHg and 8.4 +/- 4.0 mmHg, respectively. The mean +/- SEM diurnal changes in IOP after 0.005% latanoprost at 8 am once daily for the next four days were 23.3 +/- 5.0 mmHg, 25.4 +/- 2.1 mmHg, 25.7 +/- 1.7 mmHg, and 26.1 +/- 1.7 mmHg, respectively, and were significantly different from the control eye. A significant miosis also occurred starting 2 h postdrug instillation, and the resultant mean +/- SD pupil size was 1.0 +/- 0.1 mm. In the first day of the second latanoprost study, the mean +/- SEM diurnal changes in the placebo and drug eye IOPs were 11.6 +/- 3.8 mmHg, and 12.0 +/- 4.4 mmHg, respectively. For the following four days with latanoprost instilled at 8 pm, the mean +/- SEM diurnal changes in IOP in the drug eyes were 24.9 +/- 2.1 mmHg, 22.4 +/- 1.8 mmHg, 21.6 +/- 1.9 mmHg, and 26.6 +/- 2.2 mmHg, respectively. Compared to the fellow placebo eyes, the diurnal changes in IOP were significantly different. Significant changes in pupil size were similar to the IOP changes, with miosis throughout the day and return to baseline pupil size the following morning before drug instillation. In the last study, the mean +/- SEM diurnal changes in IOP for the placebo and drug eyes for the first day were 6.6 +/- 2.1 mmHg and 9.4 +/- 2.8 mmHg, respectively. For the four subsequent days with latanoprost instilled twice daily, the mean +/- SEM diurnal IOP changes were 19.6 +/- 1.5 mmHg, 19.1 +/- 1.4 mmHg, 19.9 +/- 1.7 mmHg, and 20.3 +/- 0.7 mmHg, respectively, and were significantly different from the placebo eyes. The mean changes in PS were 3.1 +/- 0.7 mm. CONCLUSION: 0.005% latanoprost instilled once daily (am or pm) as well as twice daily produces significant decreases in IOP and PS in the glaucomatous Beagle. The evening instillation of 0.005% latanoprost produced less daily fluctuations in IOP than when the drug was instilled in the morning. 0.005% latanoprost instilled twice daily produced the greatest decline in IOP with the least daily fluctuations, but longer duration miosis. PMID- 11906667 TI - Multivariate analysis of determinants of bacterial contamination of whole-blood donations. AB - BACKGROUND AND OBJECTIVES: Introduction of bacteria into blood components at the collection stage seems to be a frequent occurrence. We therefore assessed determinants of bacterial contamination of whole-blood donations to gain insight into contamination mechanisms and direct prevention. MATERIALS AND METHODS: A cross-sectional study was carried out on donors accepted for whole-blood donation in four French blood banks. Each blood bank used its own two-stage procedure for phlebotomy site preparation. Contamination was identified by culturing two 15-ml samples (collected aseptically at the outset of donation) in a BacT/Alert 240 system. Determinants were assessed by logistic regression analysis. RESULTS: Bacterial contamination, mainly by skin flora, occurred in 76 (2.2%) out of 3385 donations. Significant determinants were as follows: the blood bank (odds ratio [OR] range = 3.0-5.6, P < 0.001); lack of repetition of scrub (OR = 2.7, P = 0.032); and donor age > 35 years (OR = 1.8, P = 0.036). CONCLUSION: Systematic scrub repetition should be implemented to reduce bacterial contamination by skin flora at the collection stage. Further research is required to clarify the role of different antiseptic agents and of donor age. PMID- 11906668 TI - A case-control study of risk factors associated with human T-cell lymphotrophic virus type-I seropositivity in blood donors from Guadeloupe, French West Indies. AB - BACKGROUND AND OBJECTIVES: An age- and gender-specific distribution characterizes human T-cell lymphotrophic virus type-I (HTLV-I) seropositivity in Guadeloupe (French West Indies). Further epidemiological studies are required to identify other possible risk factors associated with this retroviral infection. MATERIALS AND METHODS: A nested case-control study was conducted between 1997 and 1999 among blood donors. A total of 102 HTLV-I-positive subjects were matched (at a ratio of 1 : 3) by gender, age (+/-5 years) and donor status (new or regular) to 306 HTLV-I-negative controls. Information was obtained through a questionnaire assessing both environmental and behavioural variables. RESULTS: Factors independently associated with HTLV-I infection included a low level of education [odds ratio (OR) 6.61, confidence interval (CI) 2.89-15.15], black ethnicity (OR 3.28, CI 1.01-10.65), two or more sex partners in the previous 3 years (OR 2.43, CI 1.16-5.10), early age at first sexual intercourse (0.84 risk reduction per additional year, CI 0.76-0.93), a history of sexually transmitted diseases (OR 2.29, CI 1.0-5.34) and positive Chlamydia serology (OR 1.95, CI 1.03-3.68). CONCLUSION: These data provide a wide spectrum of features associated with HTLV-I seropositivity, especially sexual risk factors. It strongly suggests that heterosexual intercourse is an important route of HTLV-I transmission in Guadeloupe, even among low-risk populations such as blood donors. PMID- 11906669 TI - Quality control of leucoreduced cellular blood components in France. AB - BACKGROUND AND OBJECTIVES: Since 1 April 1998, all cellular blood components in France have been leucoreduced. The current French standard is < 1 x 10(6) white blood cells (WBC) per unit with a 95% confidence that at least 97% of units will meet this standard. MATERIALS AND METHODS: Quality control (QC) data for leucoreduced cellular blood components were collected from the 41 French blood centres over a 5-month period. Conformance to the standard was determined using a non-parametric approach. RESULTS: More than 98% of the 15 286 red cell concentrates and of the 2895 platelet concentrates tested had < 1 x 10(6) WBC per unit. One filtration device gave unsatisfactory results and its use was discontinued. CONCLUSION: This QC survey shows an overall compliance with the standard. The data illustrate the practical value of identifying devices or centres with consistent QC problems. PMID- 11906670 TI - Donor selection in Japan: a trial of new criteria with predonation haemoglobin testing. AB - BACKGROUND AND OBJECTIVES: In Japan, eligibility for blood donation depends on blood specific gravity, which does not directly measure blood haemoglobin. Additionally, the criteria are not based on normal values. Therefore, we investigated the feasibility of predonation screening by using actual haemoglobin levels, and adopted a new criterion based on the normal range for men. MATERIALS AND METHODS: Using a portable device, we measured haemoglobin in 1032 prospective blood donors, then applied this method to all blood donations. The criterion for men was set at the 95th percentile of haemoglobin distribution, namely 13.0 g/dl and 13.5 g/dl, respectively, for 200-ml and 400-ml donations. That for women remained unchanged. RESULTS: The percentage of men ineligible by these criteria increased from 0.6 to 1.5%, while that of women decreased from 16.5 to 14.6%. Donors with abnormal haemoglobin levels were referred to hospitals. CONCLUSION: Predonation measurement of haemoglobin concentration, combined with the referral of those with abnormal values, provided a health benefit to that population. PMID- 11906671 TI - Evaluation of the HemoCue plasma haemoglobin analyser for assessing haemolysis in red cell concentrates during storage. AB - BACKGROUND AND OBJECTIVES: To evaluate the HemoCue plasma haemoglobin (Hb) analyser for determining supernatant Hb in red cell concentrates (RCC) during storage and compare this to a spectrophotometric method. MATERIALS AND METHODS: Samples from RCC (n = 20) at days 0, 7, 21, 35 and 42 of storage were tested by both methods for supernatant Hb. RESULTS: Results are given as median with range. There was a significant correlation (r = 0.97, P < 0.0001), but statistically significant difference between the two methods (0.72 (0.36-2.77) spectrophotometric vs. 0.70 (0.3-2.6) g/l HemoCue). However, the mean bias was negligible at 0.06 g/l. The HemoCue method gave a interassay coefficient of variation of 5.6% at 1.2 g/l and was linear to at least 20 g/l. CONCLUSION: The HemoCue plasma haemoglobin analyser is a simple, rapid, reliable and accurate method for the determination of supernatant Hb in RCC at the end of storage and compares well to established methods for supernatant Hb measurement. PMID- 11906672 TI - Recent experience with human immunodeficiency virus transmission by cellular blood products in Germany: antibody screening is not sufficient to prevent transmission. AB - BACKGROUND AND OBJECTIVES: A case of transfusion-related human immunodeficiency virus-1 (HIV-1) transmission was not detected by standard HIV antibody screening. MATERIALS AND METHODS: In a look-back procedure, the preceding donations were extensively analyzed by nucleic acid amplification technology (NAT) screening and alternative serological tests. RESULTS: The chain of infection could be demonstrated by the analysis of HIV-specific amplicon sequences from the donor and the recipient. CONCLUSION: This case report clearly indicates that the remaining risk of transfusion-related transmission of HIV could be severely reduced, not only by the use of NAT screening but even by HIV antigen screening or more sensitive HIV antibody assays. PMID- 11906673 TI - Regarding 'Rapid detection of antibodies to immunoglobulin A molecules by using the particle gel immunoassay' by Salama et al. PMID- 11906675 TI - Dosage in neutrophil transfusions. PMID- 11906677 TI - Declaration of policy regarding conflict of interest. PMID- 11906678 TI - Tobacco, nicotine, and human cognition. PMID- 11906679 TI - Women and smoking: comments on the US Surgeon General's Report. PMID- 11906680 TI - Women and smoking: a report of the Surgeon General. PMID- 11906682 TI - The subjective effects of nicotine: methodological issues, a review of experimental studies, and recommendations for future research. AB - This paper reviews findings from placebo-controlled human experimental studies of the effects of nicotine on subjective experience. Studies are grouped according to whether participants were smokers (significantly nicotine deprived, minimally nicotine deprived) or non-smokers. Within each category, studies are also grouped according to method of nicotine administration (e.g., smoked tobacco, nasal spray) and nicotine dose. This review of studies is preceded by a discussion of several methodological issues in studies of nicotine and mood. The principal findings of this review are: (1) there is a linear relationship between nicotine dose and measures of drug high (e.g., head rush, euphoria) in significantly nicotine-deprived smokers; (2) there appear to be few positive or negative main effects of nicotine on mood in minimally nicotine-deprived smokers; (3) nicotine has positive effects (e.g., increases head rush) and negative effects (e.g., increases tension) in non-smokers; (4) stronger effects of nicotine on mood emerge when individual difference variables (e.g., neuroticism) and situational contingencies (e.g., exposure to stressful stimuli) are examined. Additional studies with minimally nicotine-deprived smokers and non-smokers are needed to further specify the conditions under which nicotine affects mood and other subjective experience. The review concludes with a discussion of putative mechanisms that may mediate the interaction between nicotine and moderating variables on affect and with suggestions for future research. PMID- 11906683 TI - Tobacco smoking increases gating of irrelevant and enhances attention to relevant tones. AB - Ten adult non-smokers and 10 tobacco smokers of mixed gender were studied. The non-smokers were asked to avoid secondhand smoke; the tobacco smokers were asked to abstain from tobacco products for 6-15 h before one abstinent session and to maintain their usual smoking behavior before one smoking session. All subjects were studied twice about 1 week apart in a counterbalanced design. The tobacco smokers smoked their own brand of cigarettes in the smoking session. Auditory event-related potential recordings were begun shortly after the last puff in the smoking session. The potential recordings were repeated three times. The non smokers had no significant change in their late and long-latency auditory evoked potentials except that about 1 week later the P2 response to irrelevant tones was slightly enhanced. During tobacco abstinence, the P2 amplitude of the smokers was similar to that of the non-smokers, but was diminished after tobacco smoking. During tobacco abstinence, the P3 amplitude to relevant tones was decreased. After smoking, it was increased to that of non-smokers. Tobacco smoking also decreased the amplitude of the P2 response to frequent tones. The data support the hypothesis that tobacco smoking enhances the "protective shield" or "stimulus barrier" to irrelevant and increases attention to relevant auditory stimuli. PMID- 11906684 TI - Ethnic differences in predictors of initiation and persistence of adolescent cigarette smoking in the National Longitudinal Survey of Youth. AB - AIMS: To identify and compare predictors of adolescent smoking initiation and persistence among African American, Hispanic and White adolescents in a longitudinal national sample. DESIGN: The sample includes 1537 mother-child dyads from the National Longitudinal Survey of Youth (NLSY). Family, youth, peer and sociodemographic risk and protective factors were analyzed. FINDINGS: White adolescents reported the highest rates of smoking initiation and persistence; African Americans and Hispanics the lowest. Multivariate analyses revealed mostly common and few ethnic-specific predictors of smoking initiation and persistence. For initiation, maternal current smoking, child age, child problem behavior, and perceived peer pressure to smoke were predictive across ethnic groups; female gender and ineffective parenting were predictive among Whites only. For persistence, child age, child problem behavior and perceived scholastic competence were predictive across ethnic groups; negative mood was predictive among Whites only. CONCLUSIONS: More common than unique factors predict smoking initiation and persistence among adolescents of different ethnicity. However, the power to detect ethnicity-by-predictor interactions with respect to persistence was low. Social factors are more important for smoking initiation, whereas individual factors are more important for persistence, although child problem behaviors are common determinants both of initiation and persistence. With few exceptions, universal anti-smoking interventions should be targeted to youths of different ethnicity. PMID- 11906686 TI - Smoking and mortality following acute myocardial infarction: results from the National Registry of Myocardial Infarction 2 (NRMI 2). AB - BACKGROUND: Current smokers have lower mortality following acute myocardial infarction (AMI) than non-smokers. This is often referred to as the "smokers' paradox". Our study explored possible explanations of this phenomenon. METHODS: From the 510,044 cases of AMI in the NRMI 2 from 1 June 1994, through 30 April 1997, 297,458 cases without hospital transfer were analyzed. Characteristics and treatments of tobacco smokers and non-smokers were compared before and after age standardization. Multivariate logistic models investigated possible associations with in-hospital mortality using clinically relevant variables and interaction terms. RESULTS: Twenty-four per cent of AMI cases were current smokers. Smokers were 14 years younger than non-smokers (mean age 58 vs. 72 years, p<0.001) and had lower in-hospital mortality (8.0% vs. 16.4%, p<0.001). After age standardization, smokers were more likely than non-smokers to suffer a Q-wave type of infarction, and were less likely to have a prior history of diabetes, hypertension, AMI, angina, cardiac failure, and coronary interventions. The unadjusted odds ratio (OR) for smoking and mortality was 0.44 (95% confidence interval, CI 0.43-0.45). After adjustment for age the OR was 0.81 (95% CI 0.78 0.83). Additional adjustment for previous medical history/cardiovascular risk factors changed the OR to 0.86 (95% CI 0.83-0.89). Adjustment for additional covariates and interaction terms had little effect. CONCLUSIONS: Smokers with AMI were on average 14 years younger than non-smokers, explaining most of the apparent association of smoking with differences in presentation and treatment, and lower in-hospital mortality. The residual association of smoking and better prognosis, the "smoker's paradox", was not fully explained by measured covariates. PMID- 11906685 TI - Smoking relapse after 2 years of abstinence: findings from the VA Normative Aging Study. AB - Little is known about the risk of cigarette smoking relapse after 2 or more years of abstinence. The rates and predictors of late smoking relapse were estimated in 483 men who participated in a prospective study for up to 35 years. Subjects are participants in the VA Normative Aging Study, a prospective observational study of aging in men that began in 1963. Subjects are evaluated approximately every 3 years with physical examinations and questionnaires. Smoking, alcohol use, caffeine consumption, and socioeconomic variables were obtained by questionnaire, and weight and height were measured at clinical examinations every 3 years since 1963. Predictors of smoking relapse were identified using proportional hazards regression models. The rate of smoking relapse in the 2nd-6th years of abstinence fluctuated between 2 and 4% per year, and fell to less than 1% only after 10 years of abstinence. In multivariate regression models, coffee and alcohol consumption, and use of cigars or pipes significantly increased the risk of smoking relapse. A small risk of smoking relapse remains for at least 10 years after smoking cessation. Use of other tobacco products, coffee and alcohol increased the risk of late relapse. These findings may be useful in identifying those at highest risk for late relapse and for motivating former smokers to continue long-term abstinence. PMID- 11906687 TI - Differential effects of cigarette price on youth smoking intensity. AB - OBJECTIVES: Data from the 1992, 1993, and 1994 Monitoring the Future Surveys were used to investigate the differential effects of cigarette price on the intensity of youth cigarette smoking. METHODS: Respondents were classified into non smokers; individuals who smoked less than one cigarette per day; individuals who smoked one to five cigarettes per day; individuals who smoked one-half pack a day; and individuals who smoked one pack or more a day. A Threshold of Change Model was estimated with information on cigarette prices as the main explanatory variables. RESULTS: Dummy variables indicating medium and high prices were found to have varying effects on different levels of smoking intensity, even though higher prices were associated with lower smoking in all cases. The differences are more striking in the high-price case. The effects of higher prices are largest at the heaviest smoking levels. CONCLUSION: Cigarette prices are an effective tool to discourage youth smoking. The differential effects of cigarette price on smoking intensity warrant further investigation. PMID- 11906688 TI - Genetic and functional analysis of single nucleotide polymorphisms in the beta2 neuronal nicotinic acetylcholine receptor gene (CHRNB2). AB - The beta2-neuronal nicotinic acetylcholine receptor gene (CHRNB2) is a logical candidate for influencing smoking behavior and nicotine dependence. We discovered six single nucleotide polymorphisms (SNPs) in the CHRNB2 gene by surveying 15.4 kb of genomic sequence including a previously undescribed 3' untranslated region that extends 4.0 kb downstream of the coding region. One of the SNPs causes an amino acid substitution in exon 5, one occurs in the promoter region, one changes an intronic base, and three occur in the 3' untranslated region. The ethnically dependent allele frequencies and the marker-to-marker linkage disequilibrium patterns of five of these polymorphisms were determined. The SNPs were assayed in 743 individuals for whom information on smoking history and lifelong nicotine dependence was available. No significant associations of the individual markers or their haplotypes to smoking behavior or level of nicotine dependence were found. PMID- 11906689 TI - Pilot evaluation of a population-based health intervention for reducing use of smokeless tobacco. AB - Smokeless tobacco (ST) use has been associated with numerous negative health consequences, yet the prevalence of ST has increased dramatically since the 1970s. Young males in the military are at an elevated risk for ST use relative to the general population. Sixty active-duty male participants were identified as ST users during their annual preventive health screening and randomly assigned to minimal-contact intervention or usual care. Intervention participants were proactively contacted by phone and recruited, using a motivational interviewing style, for a cessation program consisting of a treatment manual, video, and two supportive phone calls from a cessation counselor. Sixty-five per cent (20/31) agreed to participate in the minimal-contact intervention. Three- and 6-month follow-up contacts found that the cessation rates reported by intervention participants were double those reported by participants receiving usual care (41% vs. 17% at 3 months, 37% vs. 19% at 6 months). These pilot study data suggest that proactive recruitment using a motivational interviewing approach to offer a treatment provides a good opportunity to reduce the use of ST in military settings. PMID- 11906690 TI - Hunting for excitement: NMDA receptors in Huntington's disease. AB - Excitotoxic cell death stimulated by quinolinic acid injection into the striatum has a long history of "mimicking" many aspects of motor, behavioral, and neurochemical changes observed in Huntington's disease patients. In this issue of Neuron, provide insight into the role of NMDA receptors in the cell-specific excitotoxic death observed in Huntington's disease (HD) using a HD mouse model expressing full-length mutant huntingtin (htt). PMID- 11906691 TI - Good vibrations. AB - How and where the brain forms associations between sensation and action are crucial questions in the field of cognitive neuroscience. In this issue of Neuron, add to an already impressive body of work elucidating the neural mechanisms responsible for forming categorical decisions about vibrotactile stimuli that guide behavior. Here they show that single neurons in premotor cortex represent, remarkably, what appears to be the entire temporal evolution of the decision process-from representation of the sensory stimulus, to formation of the discrimination, to the choice of behavioral output. PMID- 11906692 TI - Molecular neurobiology of human cognition. AB - Recent studies into human mental retardation syndromes have given new insights into the molecular underpinnings of human cognitive processing, in particular into mechanisms likely to contribute to learning and memory. In this minireview, we present an overview of one signal transduction cascade that has garnered attention of late in this context, the ras/ERK/CREB pathway. We focus on this cascade because of recent exciting discoveries concerning the basis of neurofibromatosis type 1 (NF1) mental retardation, which link cognitive defects in this syndrome to disruptions of ras and its intracellular targets. PMID- 11906694 TI - Fibrin inhibits peripheral nerve remyelination by regulating Schwann cell differentiation. AB - Remyelination is a critical step for functional nerve regeneration. Here we show that fibrin deposition in the peripheral nervous system after injury is a key regulator of remyelination. After sciatic nerve crush, fibrin is deposited and its clearance correlates with remyelination. Fibrin induces phosphorylation of ERK1/2 and production of p75 NGF low-affinity receptor in Schwann cells and maintains them in a nonmyelinating state, suppresses fibronectin production, and prevents synthesis of myelin proteins. In mice depleted of fibrin(ogen), remyelination of myelinated axons is accelerated due to the faster transition of the Schwann cells to a myelinating state. Regulation of fibrin clearance and/or deposition could be a key regulatory mechanism for Schwann differentiation after nerve damage. PMID- 11906693 TI - Increased sensitivity to N-methyl-D-aspartate receptor-mediated excitotoxicity in a mouse model of Huntington's disease. AB - Previous work suggests N-methyl-D-aspartate receptor (NMDAR) activation may be involved in degeneration of medium-sized spiny striatal neurons in Huntington's disease (HD). Here we show that these neurons are more vulnerable to NMDAR mediated death in a YAC transgenic FVB/N mouse model of HD expressing full-length mutant huntingtin, compared with wild-type FVB/N mice. Excitotoxic death of these neurons was increased after intrastriatal injection of quinolinate in vivo, and after NMDA but not AMPA exposure in culture. NMDA-induced cell death was abolished by an NR2B subtype-specific antagonist. In contrast, NMDAR-mediated death of cerebellar granule neurons was not enhanced, consistent with cell-type and NMDAR subtype specificity. Moreover, increased NMDA-evoked current amplitude and caspase-3 activity were observed in transgenic striatal neurons. Our data support a role for NR2B-subtype NMDAR activation as a trigger for selective neuronal degeneration in HD. PMID- 11906695 TI - Neuropilin-2 mediates axonal fasciculation, zonal segregation, but not axonal convergence, of primary accessory olfactory neurons. AB - The mechanisms that underlie axonal pathfinding of vomeronasal neurons from the vomeronasal organ (VNO) in the periphery to select glomeruli in the accessory olfactory bulb (AOB) are not well understood. Neuropilin-2, a receptor for secreted semaphorins, is expressed in V1R- and V3R-expressing, but not V2R expressing, postnatal vomeronasal neurons. Analysis of the vomeronasal nerve in neuropilin-2 (npn-2) mutant mice reveals pathfinding defects at multiple choice points. Vomeronasal sensory axons are severely defasciculated and a subset innervates the main olfactory bulb (MOB). While most axons of V1R-expressing neurons reach the AOB and converge into distinct glomeruli in stereotypic locations, they are no longer restricted to their normal anterior AOB target zone. Thus, Npn-2 and candidate pheromone receptors play distinct and complementary roles in promoting the wiring and patterning of sensory neurons in the accessory olfactory system. PMID- 11906696 TI - Novel modulation of neuronal nicotinic acetylcholine receptors by association with the endogenous prototoxin lynx1. AB - We previously identified lynx1 as a neuronal membrane molecule related to snake alpha-neurotoxins able to modulate nAChRs. Here, we show that lynx1 colocalizes with nAChRs on CNS neurons and physically associates with nAChRs. Single-channel recordings show that lynx1 promotes the largest of three current amplitudes elicited by ACh through alpha(4)beta(2) nAChRs and that lynx1 enhances desensitization. Macroscopic recordings quantify the enhancement of desensitization onset by lynx1 and further show that it slows recovery from desensitization and increases the EC(50). These experiments establish that direct interaction of lynx1 with nAChRs can result in a novel type of functional modulation and suggest that prototoxins may play important roles in vivo by modulating functional properties of their cognate CNS receptors. PMID- 11906697 TI - Synaptic mechanisms underlie nicotine-induced excitability of brain reward areas. AB - A single nicotine exposure increases dopamine levels in the mesolimbic reward system for hours, but nicotine concentrations experienced by smokers desensitize nAChRs on dopamine neurons in seconds to minutes. Here, we show that persistent modulation of both GABAergic and glutamatergic synaptic transmission by nicotine can contribute to the sustained increase in dopamine neuron excitability. Nicotine enhances GABAergic transmission transiently, which is followed by a persistent depression of these inhibitory inputs due to nAChR desensitization. Simultaneously, nicotine enhances glutamatergic transmission through nAChRs that desensitize less than those on GABA neurons. The net effect is a shift toward excitation of the dopamine reward system. These results suggest that spatial and temporal differences in nicotinic receptor activity on both excitatory and inhibitory neurons in reward areas coordinate to reinforce nicotine self administration. PMID- 11906699 TI - AII (Rod) amacrine cells form a network of electrically coupled interneurons in the mammalian retina. AB - AII (rod) amacrine cells in the mammalian retina are reciprocally connected via gap junctions, but there is no physiological evidence that demonstrates a proposed function as electrical synapses. In whole-cell recordings from pairs of AII amacrine cells in a slice preparation of the rat retina, bidirectional, nonrectifying electrical coupling was observed in all pairs with overlapping dendritic trees (average conductance approximately 700 pS). Coupling displayed characteristics of a low-pass filter, with no evidence for amplification of spike evoked electrical postsynaptic potentials by active conductances. Coincidence detection, as well as precise temporal synchronization of subthreshold membrane potential oscillations and TTX-sensitive spiking, was commonly observed. These results indicate a unique mode of operation and integrative capability of the network of AII amacrine cells. PMID- 11906698 TI - Distinct NMDA receptors provide differential modes of transmission at mossy fiber interneuron synapses. AB - Dentate gyrus granule cells innervate inhibitory interneurons via a continuum of synapses comprised of either Ca(2+)-impermeable (CI) or Ca(2+)-permeable (CP) AMPA receptors. Synapses at the extreme ends of this continuum engage distinct postsynaptic responses, with activity at CI synapses being strongly influenced by NMDA receptor activation. NMDARs at CI synapses have a lower NR2B subunit composition and a higher open probability, which generate larger amplitude and more rapid EPSCs than their CP counterparts. A novel form of NMDAR-dependent long term depression (iLTD) is associated with CI-mossy fiber synapses, whereas iLTD at CP synapses is dependent on Ca(2+)-permeable AMPA receptor activation. Induction of both forms of iLTD required elevation of postsynaptic calcium. Thus mossy fibers engage CA3 interneurons via multiple synapse types that will act to expand the computational repertoire of the mossy fiber-CA3 network. PMID- 11906700 TI - Cellular mechanisms of the slow (<1 Hz) oscillation in thalamocortical neurons in vitro. AB - The slow (<1 Hz) rhythm is a defining feature of the electroencephalogram during sleep. Since cortical circuits can generate this rhythm in isolation, it is assumed that the accompanying slow oscillation in thalamocortical (TC) neurons is largely a passive reflection of neocortical activity. Here we show, however, that by activating the metabotropic glutamate receptor (mGluR), mGluR1a, cortical inputs can recruit intricate cellular mechanisms that enable the generation of an intrinsic slow oscillation in TC neurons in vitro with identical properties to those observed in vivo. These mechanisms rely on the "window" component of the T type Ca(2+) current and a Ca(2+)-activated, nonselective cation current. These results suggest an active role for the thalamus in shaping the slow (<1 Hz) sleep rhythm. PMID- 11906701 TI - Temporal evolution of a decision-making process in medial premotor cortex. AB - The events linking sensory discrimination to motor action remain unclear. It is not known, for example, whether the motor areas of the frontal lobe receive the result of the discrimination process from other areas or whether they actively participate in it. To investigate this, we trained monkeys to discriminate between two mechanical vibrations applied sequentially to the fingertips; here subjects had to recall the first vibration, compare it to the second one, and indicate with a hand/arm movement which of the two vibrations had the higher frequency. We recorded the activity of single neurons in medial premotor cortex (MPC) and found that their responses correlate with the diverse stages of the discrimination process. Thus, activity in MPC reflects the temporal evolution of the decision-making process leading to action selection during this perceptual task. PMID- 11906702 TI - A network representation of response probability in the striatum. AB - The striatum of the basal ganglia is considered a key structure in the learning circuitry of the brain. To analyze neural signals that underlie striatal plasticity, we recorded from an identifiable class of striatal interneurons as macaque monkeys underwent training in a range of conditioning and non-associative learning paradigms, and recorded eyeblink electromyographs as the measure of behavioral response. We found that the responses of these striatal interneurons were modifiable under all training conditions and that their population responses were tightly correlated with the probability that a given stimulus would evoke a behavioral response. Such a network signal, proportional to current response probability, could be crucial to the learning and decision functions of the basal ganglia. PMID- 11906703 TI - The role of the amygdala in signaling prospective outcome of choice. AB - Can brain activity reveal a covert choice? Making a choice often evokes distinct emotions that accompany decision processes. Amygdala has been implicated in choice behavior that is guided by a prospective negative outcome. However, its specific involvement in emotional versus cognitive processing of choice behavior has been a subject of controversy. In this study, the human amygdala was monitored by functional magnetic resonance imaging (fMRI) while subjects were playing in a naturalistic choice paradigm against the experimenter. In order to win, players had to occasionally choose to bluff their opponent, risk "getting caught," and suffer a loss. A critical period, when choice has been made but outcome was still unknown, activated the amygdala preferentially following the choice that entailed risk of loss. Thus, the response of the amygdala differentiated between subject's covert choice of either playing fair or foul. These results support a role of the amygdala in choice behavior, both in the appraisal of inherent value of choice and the signaling of prospective negative outcomes. PMID- 11906704 TI - The role of angiotensin II in cognition and behaviour. AB - Polymorphisms of the renin-angiotensin system are associated with cardiovascular disorders, possibly as a consequence of increased brain angiotensin II activity. Within the brain, angiotensin controls blood pressure, fluid balance and hormone secretion; it also influences behaviour: reduction of central angiotensin function has both antidepressant-like and axiolytic-like actions. Evidence concerning the role of the renin-angiotensin system in learning and memory is contradictory, although more studies support the proposal that angiotensin reduces cognitive function. Studies of renin-angiotensin system genotype and psychological status have suggested an association between the angiotensin converting enzyme deletion allele and age related cognitive decline, but a greater prevalence of the insertion allele in Alzheimer's disease. The deletion allele has also been associated with depressive illness, as has the M allele of the angiotensinogen gene although other studies have failed to replicate these findings. The role of the brain renin-angiotensin system in human psychopathology remains to be fully explored. PMID- 11906705 TI - Effect of glutathione S-transferases on the survival of patients with acute myeloid leukaemia. AB - The objective of the study was to investigate the effect of genetic polymorphisms in glutathione S-transferases (GST) on the survival of acute myeloid leukaemia patients receiving adriamycin induction therapy. A total of 89 patients were included in the study. Patients who carried at least one GSTM1 allele had trend towards a better survival (mortality rate ratio (RR) 0.588; 95% CI 0.334-1.036) than GSTM1*0/0 patients. However, at low accumulated adriamycin dose, GSTM1*0/0 cases had a better survival than people expressing the gene (RR=6.1; 95% CI=1.2 11.0). The GSTT1 and GSTP1 genotype did not influence the survival in any of the groups. PMID- 11906706 TI - Methotrexate-induced apoptosis in hepatocytes after partial hepatectomy. AB - To investigate apoptosis induced by methotrexate in hepatocytes in vivo, rats received a single injection of methotrexate immediately after partial hepatectomy and apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) and gel electrophoresis of DNA. Characteristic DNA fragmentation was obvious at 2 h and peaked at 4 h after partial hepatectomy with methotrexate injection. TUNEL-positive staining was observed in nuclei and nuclear fragments of hepatocytes in the methotrexate-injected liver (partial hepatectomy with methotrexate), with negligible background staining in the control (partial hepatectomy only) and in the methotrexate-injected normal (normal with methotrexate) rat liver. The involvement of the c-Jun N-terminal kinase (JNK) activator protein 1 (AP-1) pathway and p53 in apoptosis was also examined. The activity of JNK increased at 15 min and peaked at 1 h after partial hepatectomy. This increase was repressed by methotrexate injection. Western blot analysis showed that the levels of c-Fos and c-Jun protein expression, which increased at 1 h after partial hepatectomy, were also reduced by methotrexate. The levels of p53 protein were markedly increased after partial hepatectomy with methotrexate injection. The increase in p53 protein was followed by an up regulation of p21(WAF1/CIP1) protein at 2 h after partial hepatectomy. These results suggested that the inhibition of the JNK-AP-1 pathway and concurrent up regulation of p53 and p21(WAF1/CIP1) were involved in hepatocyte apoptosis induced by partial hepatectomy with methotrexate. PMID- 11906707 TI - Identification and characterisation of functional bombesin receptors in human astrocytes. AB - Reverse transcription polymerase chain reaction (RT-PCR) demonstrated the presence of bombesin BB2 receptor mRNA but not bombesin BB1 receptor or bombesin BB3 receptor mRNA in cultured human astrocytes. Neuromedin C hyperpolarised human astrocytes in whole-cell current and voltage clamp recordings and increased the intracellular free Ca(2+) ion concentration ([Ca(2+)](i)) in single astrocytes. Treatment with neuromedin C caused larger and more frequent increases in [Ca(2+)](i) than those triggered by neuromedin B, with 96% and 78% of cells responding, respectively. The stimulatory effects of neuromedin C were inhibited significantly by treatment with U73122 or the bombesin BB2 receptor antagonist [D Phe(6), des-Met(14)]bombesin-(6-14) ethylester. A Fluorometric Imaging Plate Reader (FLIPR) was used to measure [Ca(2+)](i) in cell populations. Neuromedin C was approximately 50-fold more potent than neuromedin B in elevating [Ca(2+)](i) in astrocytes and Chinese hamster ovary (CHO) cells expressing human bombesin BB2 receptors (hBB2-CHO). However, in CHO cells expressing the bombesin BB1 receptor hBB1-CHO, neuromedin B was 32-fold more potent than neuromedin C. [D-Phe(6), des Met(14)]bombesin-(6-14) ethylester was a partial agonist in hBB1-CHO cells (E(max)=55%) but was a noncompetitive antagonist in both hBB2-CHO cells and astrocytes. These studies report the first identification of functional bombesin receptors on cultured human astrocytes and have demonstrated that the bombesin BB2 receptor contributes significantly to astrocyte physiology. PMID- 11906708 TI - Isobolographic analysis of non-depolarising muscle relaxant interactions at their receptor site. AB - Administration of certain combinations of non-depolarising muscle relaxants produces greater than expected neuromuscular blockade. Synergistic effects may be explained by drug interactions with the postsynaptic muscle nicotinic acetylcholine receptor. To investigate this hypothesis, the adult mouse muscle nicotinic acetylcholine receptor (alpha(2)beta delta epsilon) was heterologously expressed in Xenopus laevis oocytes and activated by the application of acetylcholine (10 microM). The effects of five individually applied muscle relaxants and six combinations of structurally similar and dissimilar compounds were studied. Drug combinations containing equipotent concentrations of two agents were tested and dose-response curves were determined. All compounds tested alone and in combination produced rapid and readily reversible, concentration dependent inhibition. Isobolographic and fractional analyses indicated additive interactions for all six tested combinations. These findings suggest that synergistic neuromuscular blocking effects, observed for the administration of certain combinations of muscle relaxants, do not result from purely postsynaptic binding events at the muscle nicotinic acetylcholine receptor, but rather from differential actions on pre- and postsynaptic sites. PMID- 11906709 TI - Intracellular cAMP increases during the positive inotropism induced by androgens in isolated left atrium of rat. AB - Molecular interactions of androgens with the plasma membrane may produce rapid cardiovascular effects that cannot be explained by the classic genomic mechanisms. In this sense, 5 alpha- and 5 beta-dihydrotestosterone-induced an acute positive inotropic effect in isolated left atrium of rat, an effect which may be due to cAMP-dependent mechanisms. To prove this, intracellular levels of cAMP, after exposure to androgens in the organ bath, and binding to beta(1) adrenoceptors were evaluated. After a 4-min exposure, 5 alpha- and 5 beta dihydrotestosterone increased cAMP levels from 3.83+/-0.61 to 6.15+/-1.1 and 11.18+/-2.4 pmol cAMP/mg of protein, respectively. These increases were inhibited by atenolol and not modified by treatment of the rats with reserpine. The androgen-induced cAMP increase seems to be produced via an extracellular interaction, because positive inotropism and raised levels of cAMP were produced by 5 alpha-dihydrotestosterone conjugated with bovine serum albumin (BSA). In addition, it is independent of beta(1)-adrenoceptor activation, because neither androgen displaced [(3)H]dihydroalprenolol binding. Therefore, the androgens induced a positive inotropic effect via a postsynaptic effect that increases intracellular levels of cAMP. This effect is modulated by transcriptional mechanisms or by a protein with a short half-life. PMID- 11906710 TI - AMPA-induced Ca(2+) influx in cultured rat cortical nonpyramidal neurones: pharmacological characterization using fura-2 microfluorimetry. AB - Immunocytochemical and Co(2+) uptake studies revealed that in primary cultures of rat cortical neurones, the majority of neurones are gamma-aminobutyric acid (GABA) immunopositive and can express Ca(2+)-permeable alpha-amino-3-hydroxy-5 methyl-4-isoxazole-propionic acid (AMPA) receptors. By fura-2 microfluorimetry, it was shown that the stimulation with the selective agonist (S)-AMPA (0.3-300 microM) induced a concentration-dependent but cell-variable increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) (EC(50) value 7.4 microM) in more than 80% of the medium-sized multipolar neurones studied. The AMPA-induced rise in [Ca(2+)](i) seems to be due to Ca(2+) entry through AMPA receptor channels, because the response was abolished in Ca(2+)-free solution and by AMPA receptor selective antagonists, but was not significantly influenced by cyclopiazonic acid, an inhibitor of the endoplasmatic Ca(2+)-ATPase, by selective N-methyl-D-aspartic acid (NMDA) receptor antagonists, as well as the Na(+) channel blocker tetrodotoxin and the majority of tested Ca(2+) channel blockers. In conclusion, the results indicate that the cerebral cortical neurones in culture represent mostly GABAergic interneurone-like cells and the majority of them possess Ca(2+)-permeable AMPA receptors, important for intracellular signal transduction and neuronal plasticity. PMID- 11906711 TI - Na(+) channel effects of remacemide and desglycinyl-remacemide in rat cortical synaptosomes. AB - The effects of the novel anticonvulsant, remacemide hydrochloride and its active metabolite, desglycinyl-remacemide, on veratridine-induced Na(+) influx in rat cortical synaptosomes were investigated and compared to established Na(+) channel blocking antiepileptic drugs. Remacemide and desglycinyl-remacemide reduced veratridine-stimulated Na(+) influx to 30.7% (IC(50)=160.6 microM) and 13.2% (IC(50)=85.1 microM) of control, respectively. Carbamazepine, phenytoin and lamotrigine similarly reduced Na(+) influx to 20.1% (IC(50)=325.9 microM), 79.8% and 27.9% (IC(50)=23.0 microM) of control, respectively. Resting internal Na(+) concentrations were significantly increased by desglycinyl-remacemide (1 and 10 microM) and, conversely, decreased by desglycinyl-remacemide and carbamazepine (both 1000 microM). These studies support previous electrophysiological investigations, which suggest that remacemide and desglycinyl-remacemide exert their antiepileptic effects, at least in part, by an inhibitory action on voltage gated Na(+) channels. Desglycinyl-remacemide may have an additional action on Na(+) homeostasis that merits further exploration. PMID- 11906712 TI - Inhibition of nitric oxide-dependent activation of soluble guanylyl cyclase by the antimalarial drug, artemisinin. AB - The influence of artemisinin on the activity of human platelet soluble guanylyl cyclase was investigated. Artemisinin (0.1-100 microM) had no effect on the basal activity of the enzyme. Artemisinin inhibited in a concentration-dependent manner the sodium nitroprusside-induced activation of human platelet guanylyl cyclase with an IC(50) value 5.6 microM. Artemisinin (10 microM) also inhibited (by 71+/ 4.0%) the activation of the enzyme by the thiol-dependent nitric oxide (NO) donor, the derivative of furoxan, 3,4-dicyano-1,2,5-oxadiazole 2-oxide (10 microM), but did not influence the stimulation of soluble guanylyl cyclase by protoporphyrin 1X. Inhibition of guanylyl cyclase activation by NO donors but not by protoporphyrin 1X represents a possible additional mechanism of the pharmacological action of this drug. PMID- 11906713 TI - Phencyclidine increases vesicular dopamine uptake. AB - Phencyclidine (PCP) rapidly (within 1 h) increased vesicular dopamine uptake and binding of the vesicular monoamine transporter-2 (VMAT-2) ligand, dihydrotetrabenazine. Uptake returned to basal values 3 h in the striatum after a high-dose administration of this drug (15 mg/kg i.p.). In contrast, a similar pretreatment with another non-competitive NMDA receptor antagonist, dizocilpine;([5R,10S]-[+]-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10 imine; MK-801; 1 mg/kg, i.p.), was without effect on vesicular dopamine uptake. Pretreatment with the dopamine D2 receptor antagonist, eticlopride, blocked the increase in vesicular dopamine uptake caused by PCP administration. These data demonstrate a heretofore unreported mechanism that may contribute to the ability of PCP to influence dopamine neuronal function and exert its pharmacological effects. PMID- 11906714 TI - Sodium nitroprusside stimulates L-DOPA release from striatal tissue through nitric oxide and cGMP. AB - The effects of the nitric oxide (NO) donor, sodium nitroprusside, on L-DOPA and dopamine release from striatal tissue were evaluated using a static incubation system in which the striatal tissue released between three and six times more L DOPA than DA, although the DA content was four times higher than that of L-DOPA. Sodium nitroprusside stimulated L-DOPA release in a time- and concentration dependent (25, 50 and 100 microM) manner. This effect was not due to an increase in L-DOPA synthesis because sodium nitroprusside did not modify the tyrosine hydroxylase activity of striatal tissue. DA release was also stimulated by sodium nitroprusside but it required a higher concentration (500 microM) and longer incubation (60 min). Neither basal nor sodium nitroprusside-stimulated L-DOPA release was influenced by Ca(2+) deprivation (EGTA 5 mM) and/or the presence of nitrendipine (1 microM), a blocker Ca(2+) channel, in the incubation medium. However, cGMP (1 mM) increased L-DOPA release, and the soluble guanylate cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ) (5 microM), partially blunted the stimulatory effect of sodium nitroprusside 100 microM. In addition, the presence of certain scavengers of free radicals, such as uric acid (300 microM) or melatonin (300 microM) but not of superoxide dismutase (1000 UI/ml) or salicylic acid (300 microM), completely blocked sodium nitroprusside (100 microM)-induced L-DOPA release. These results show that NO stimulates L-DOPA release from striatal tissue by an apparently Ca(2+)-independent mechanism, mediated by cGMP but also by peroxynitrite. PMID- 11906715 TI - Pharmacological evidence for the activation of K(+) channels by diclofenac. AB - The involvement of K(+) channels in the antinociceptive action of diclofenac was assessed in the formalin test. Local administration of diclofenac produced a dose dependent antinociceptive effect due to a local action because drug administration in the contralateral paw was ineffective. Pretreatment of the injured paw with glibenclamide and tolbutamide (ATP-sensitive K(+) channel inhibitors), charybdotoxin and apamin (large- and small-conductance Ca(2+) activated K(+) channel blockers, respectively), 4-aminopyridine or tetraethylammonium (voltage-dependent K(+) channel inhibitors) prevented diclofenac-induced antinociception. Given alone, K(+) channel inhibitors did not modify formalin-induced nociceptive behavior. Pinacidil (an ATP-sensitive K(+) channel opener) also produced antinociception which was blocked by glibenclamide. The peripheral antinociceptive effect of morphine (positive control) was blocked by glibenclamide and 4-aminopyridine but not by charybdotoxin or apamin. The results suggest that the peripheral antinociceptive effect of diclofenac may result from the activation of several types of K(+) channels, which may cause hyperpolarization of peripheral terminals of primary afferents. PMID- 11906716 TI - Tacrolimus, a specific inhibitor of calcineurin, modifies the locomotor activity of quinpirole, but not that of SKF82958, in male rats. AB - In the present study, we examined the effect of tacrolimus, a specific inhibitor of calcineurin, on the locomotor activity elicited by the selective dopamine D(1) receptor agonist (+/-) 6-chloro-7,8-dyhydroxy-3allyl-1-phenyl-2,3,4,5-tetra-hydro 1H-benzazepine (SKF82958) and the dopamine D(2)/D(3) receptor agonist quinpirole, in male Wistar rats. Tacrolimus (0.5, 1, 2 or 5 mg/kg, i.p.) alone had no significant effect on basal locomotor activity. The dose-related increase in locomotor activity produced by the administration of SKF82958 (0.1, 1 or 5 mg/kg, i.p.) was not significantly altered by 2 mg/kg of tacrolimus. In addition, the increase in locomotor activity produced by 1 mg/kg of SKF82958 was not significantly altered by tacrolimus (0.5, 1, 2 or 5 mg/kg, i.p.). The administration of quinpirole (0.1, 0.25, 0.5, 1 or 3 mg/kg, i.p.) produced a biphasic response, with the minimum and maximal increase in locomotor activity occurring at 0.1 and 1 mg/kg, respectively. The pretreatment of 2 mg/kg of tacrolimus, compared to vehicle-treated animals, significantly lowered the dose of quinpirole that produce a maximal effect on locomotor activity from 1 to 0.5 mg/kg but did not significantly alter the minimum response. The increase in locomotor activity produced by 0.5 mg/kg of quinpirole was significantly potentiated by 0.5, 1, 2 or 5 mg/kg of tacrolimus compared to vehicle-treated animals. Our results suggest that calcineurin may play a role in the alteration of locomotor activity produced by dopamine D(2)/D(3) receptors, but not dopamine D(1) receptors. PMID- 11906717 TI - Antagonism of alpha 3 beta 4 nicotinic receptors as a strategy to reduce opioid and stimulant self-administration. AB - The iboga alkaloid ibogaine and the novel iboga alkaloid congener 18 methoxycoronaridine are putative anti-addictive agents. Using patch-clamp methodology, the actions of ibogaine and 18-methoxycoronaridine at various neurotransmitter receptor ion-channel subtypes were determined. Both ibogaine and 18-methoxycoronaridine were antagonists at alpha 3 beta 4 nicotinic receptors and both agents were more potent at this site than at alpha 4 beta 2 nicotinic receptors or at NMDA or 5-HT(3) receptors; 18-methoxycoronaridine was more selective in this regard than ibogaine. In studies of morphine and methamphetamine self-administration, the effects of low dose combinations of 18 methoxycoronaridine with mecamylamine or dextromethorphan and of mecamylamine with dextromethorphan were assessed. Mecamylamine and dextromethorphan have also been shown to be antagonists at alpha 3 beta 4 nicotinic receptors. All three drug combinations decreased both morphine and methamphetamine self-administration at doses that were ineffective if administered alone. The data are consistent with the hypothesis that antagonism at alpha 3 beta 4 receptors is a potential mechanism to modulate drug seeking behavior. 18-Methoxycoronaridine apparently has greater selectivity for this site than other agents and may be the first of a new class of synthetic agents acting via this novel mechanism to produce a broad spectrum of anti-addictive activity. PMID- 11906718 TI - Mechanism underlying gamma-aminobutyric acid-induced antihypertensive effect in spontaneously hypertensive rats. AB - We examined the effects of gamma-aminobutyric acid (GABA) on the blood pressure in spontaneously hypertensive rats and normotensive Wistar-Kyoto rats. A single oral administration (0.5 mg/kg) significantly lowered the systolic blood pressure in spontaneously hypertensive rats, but not in normotensive rats. In the mesenteric arterial bed, the perivascular nerve stimulation-induced increase in perfusion pressure and noradrenaline release were significantly inhibited by GABA in spontaneously hypertensive rats, but not in normotensive rats, and attenuated by the selective GABA(B) receptor agonist, baclofen, but not by the selective GABA(A) receptor agonist muscimol. The inhibitory effects of GABA on the perivascular nerve stimulation-induced increase in perfusion pressure and noradrenaline release were completely antagonized by the selective GABA(B) receptor antagonist, saclofen, but not by the selective GABA(A) receptor antagonist, bicuculline. These results suggest that, in spontaneously hypertensive rats, GABA has an antihypertensive effect due to its inhibition of noradrenaline release from sympathetic nerves in the mesenteric arterial bed via presynaptic GABA(B) receptors. PMID- 11906719 TI - Long-term potentiation in rat prefrontal slices facilitated by phased application of dopamine. AB - We have previously shown that coupling bath application of dopamine with 50 Hz tetani induces long-term depression in rat prefrontal slices [Neuroscience 85 (1998) 669]. Here, we report a reliable protocol for inducing long-term potentiation in the same preparation. Long-term potentiation was induced by the same dopamine-tetani coupling protocol when the coupling was preceded (approximately 30 min) by a single bath application of dopamine. We suggest that metaplastic processes triggered by the first application of dopamine might underlie the LTP induction. PMID- 11906720 TI - Amiloride-sensitive epithelial sodium channel subunits are expressed in human and mussel immunocytes. AB - In this study, we examined the expression of epithelial Na(+) channel (ENaC) subunits in human peripheral blood lymphocytes, human lymph nodes and molluscan immunocytes using non-radioactive in situ hybridization. The results showed that T lymphocytes express the ENaC gamma subunit mRNA, and B lymphocytes the ENaC beta subunit mRNA. Yet, the alpha subunit mRNA was not detected in either cell type. In molluscan immunocytes, all three homologous ENaC subunit mRNAs are present, and these data were also confirmed by RT-PCR and sequencing of the PCR products. These findings show evolutionary conservation of the expression of ENaC subunits in immunocytes of invertebrates to vertebrates. The observed differential expression patterns of ENaC subunits suggest that ENaC function may be regulated differentially in different types of human lymphocytes. PMID- 11906721 TI - Conservation of the modular structure of complement factor I through vertebrate evolution. AB - Mammalian complement factor I plays pivotal roles in the regulation of complement activation and generation of important biological activities from C3. The evolutionary origin of factor I has been unclear except with regard to the molecular cloning of factor I from amphibian Xenopus. Here, we report the identification and characterization of factor I cDNA from the liver of the banded houndshark. The deduced amino acid sequence of shark factor I showed a modular organization that was completely identical to that of mammalian factor I, suggesting the functional conservation of factor I throughout vertebrate evolution. Functionally important amino acid residues such as the basic residues at the processing site and the residues at the active site of the serine protease domain are conserved. Repeated sequences composed of 16 amino acids were inserted at a site between the leader peptide and the factor I/membrane attacking complex module in the shark factor I. This repeat is missing from mammalian and amphibian factor I, and the biological significance of the sequence, if any, is not clear at the moment. There was only one copy of the shark factor I gene, and Northern blotting analysis showed that the shark factor I gene was expressed only in the liver among several organs tested. While the lack of functional data does not exclude the possibility that factor I could have a different function, all these facts, together with the earlier reported data suggest the existence of a well developed complement system in cartilaginous fish. PMID- 11906722 TI - Cloning of a Salmo salar interleukin-1 receptor-like cDNA. AB - The interleukin-1 receptor/toll-like receptor (IL-1R/TLR) superfamily, defined by a cytosolic Toll/IL-1R (TIR) signalling domain, participates in host responses to injury and infection. We describe in this study the cloning of a cDNA encoding a Salmo salar interleukin-1 receptor-like protein (SalIL-1RLP). SalIL-1RLP comprises a potential signal peptide, three extracellular immunoglobulin domains, a short transmembrane region and an intracellular region that contains the TIR domain. The predicted amino acid sequence of SalIL-1RLP displays 43-44% similarities and 31% identities to chicken and human IL-1RI sequences. Within the intracellular region, SalIL-1RLP displays highest similarity (59%) and identity (46%) to the chicken IL-1RI sequence. Two different 5' distal UTRs were identified among six salmon IL-1RLP clones. The six clones, however, displayed identical 5' proximal UTRs, coding regions and 3' UTRs. SalIL-1RLP expression is induced in liver, head kidney, spleen and gills upon injection of salmon with bacterial lipopolysaccharide. Sequence comparisons, protein domain structures, expression patterns and phylogenetic analyses indicate that SalIL-1RLP is most closely related to type I interleukin-1 receptors and interleukin-1 receptor related proteins. PMID- 11906723 TI - Identification and analysis of an interleukin 8-like molecule in rainbow trout Oncorhynchus mykiss. AB - An interleukin 8 (IL-8) homologue has been identified in the rainbow trout Oncorhynchus mykiss. The transcript contains an open reading frame of 294 nucleotides that translates into a 97 amino acid putative peptide, with 5' and 3' untranslated regions (UTR) of 171 and 453 nucleotides, respectively. As with previously sequenced lamprey and flounder genes, the trout amino acid sequence lacks the typical ELR motif upstream of the first pair of cysteines, where DLR is present. The trout IL-8 gene contains four exons divided by three short introns of 341, 247 and 292bp, and occupies 1824bp of genomic DNA. RT-PCR reveals a low level constitutive expression of the IL-8 homologue in many tissues, including spleen, heart, liver, head kidney and gill. Expression was not detectable in the brain. Whilst no apparent affect of lipopolysaccharide (LPS) on IL-8 expression was observed in vivo, stimulation of a trout macrophage cell line (RTS-11) with either LPS or poly I:C did result in clear up-regulation of IL-8 expression, detectable by RT-PCR and Northern blot analysis. PMID- 11906724 TI - Induction of nitric oxide and respiratory burst response in activated goldfish macrophages requires potassium channel activity. AB - Potassium channel activity is important for modulating mammalian macrophage antimicrobial functions. The involvement of potassium channels in mediation of immune cell function in lower vertebrates, such as teleost, has not been explored. Since relatively little is known about the types of potassium channels present in fish macrophages, pharmacological blockers with broad ranges of activity were tested: 4-aminopyridine (4-AP), quinine, and tetraethylammonium chloride (TEA). The potassium channel blockers inhibited reactive nitrogen intermediates (RNI) and reactive oxygen intermediates (ROI) production by goldfish macrophages activated with bacterial lipopolysaccharide (LPS) and/or macrophage activating factor (MAF)-containing supernatants. Quinine was the most potent inhibitor with an IC(50) of 50 microM, while the other blockers, 4-AP and TEA, had IC(50) of 1.2 and 0.6mM, respectively. A reversible depolarization of the goldfish macrophage plasma membrane potential (Vm) was observed following treatments with potassium channel blockers, and was related to transcriptional changes in the inducible nitric oxide synthase gene (iNOS). Down-regulation of antimicrobial activities and depolarization of the goldfish macrophage plasma membrane were not a consequence of reduced cell number or viability, suggesting that potassium channels are required for generation of appropriate goldfish macrophage antimicrobial functions. PMID- 11906725 TI - Activation of carp leukocytes by a galactose-binding protein from Aphanomyces piscicida. AB - This study demonstrated that a galactose-binding protein (GBP) produced by a fish pathogenic water mold, Aphanomyces piscicida, activates carp leukocytes. Leukocytes were separated from the head kidney and peripheral blood using Percoll density centrifugation. A flow cytometric analysis revealed that GBP binds with many cells and a variety of cell types including lymphocytes, granulocytes and thrombocytes. Intracellular calcium flux of the peripheral blood leukocytes induced by stimulation with GBP was confirmed by counting the fluo-3 loaded cells whose fluorescence increased after the stimulation using flow cytometry. The percentage of cells in which a calcium flux was induced peaked 1 min after the stimulation. Approximately 6% of the cells specifically responded 1 min after the stimulation. The proliferation response was determined by the level of BrdU uptake by the leukocytes after the stimulation. Cell proliferation was observed 2, 4 and 6 days after stimulation with GBP. The expression of cytokines IL-1beta and TGF-beta1 in the peripheral blood leukocytes, after the stimulation was evaluated by a semi-quantitative reverse-transcriptase polymerase chain reaction. Increased expression of IL-1beta was observed 4h after stimulation with GBP. Variation of TGF-beta1 expression under the same conditions was not observed. The kinetics of intracellular calcium flux and the level of IL-1beta expression induced by GBP stimulation were different from those induced by phytohemagglutinin stimulation. These results confirmed that GBP is a pathogenic microbial component that can induce cell activation. GBP seems to induce the inflammatory response observed in the Aphanomyces infection. PMID- 11906726 TI - Activity of antimicrobial skin peptides from ranid frogs against Batrachochytrium dendrobatidis, the chytrid fungus associated with global amphibian declines. AB - Accumulating evidence suggests that a chytrid fungus, Batrachochytrium dendrobatidis, is responsible for recent declines in amphibian populations in Australia, Central America, Europe, and North America. Because the chytrid infects the keratinized epithelium of the skin, we investigated the possible role of antimicrobial peptides produced in the skin as inhibitors of infection and growth. We show here that 10 peptides representing eight families of peptides derived from North American ranid frogs can effectively inhibit growth of this chytrid. The peptides are members of the ranatuerin-1, ranatuerin-2, esculentin 1, esculentin-2, brevinin-2, temporin, palustrin-3, and ranalexin families. All the tested peptides inhibit growth of mature fungal cells at concentrations above 25 microM, and some of them inhibit at concentrations as low as 2 microM. A comparison of the sensitivity of infectious zoospores with that of mature cells showed that the zoospores are inhibited at significantly lower concentrations of peptides. To determine whether cold temperature interferes with the inhibitory effects of these peptides, we tested their effectiveness at both 22 and 10 degrees C. Although the peptides inhibit at both temperatures, they appear to be more effective against zoospores at the lower temperature. These results suggest that the ranid frogs have, within their repertoire of antimicrobial substances, a number of skin peptides that should be a deterrent to chytrid infection. This may provide some natural resistance to infection, but if environmental factors inhibit the synthesis and release of the skin peptides, the pathogen could gain the advantage. PMID- 11906727 TI - Diverse expression of the K-1 antigen by cortico-medullary and reticular epithelial cells of the bursa of Fabricius in chicken and guinea fowl. AB - The immunocytochemical study of the K-1 monoclonal antibody indicates that the epithelial components of the bursa of Fabricius of the chicken and guinea fowl express the K-1 positive molecule. During embryogenesis, the K-1 antigen expression appears together with the bud-formation. As the number of B cells increases in the developing follicle, the K-1 expression gradually diminishes in the medullary reticular epithelial cells and completely ceases by hatching, which suggests that the molecule is developmentally regulated. After hatching, the expression of the molecule is restricted to the sealing off zone of the lymphoepithelial or medullary region of the follicle: i.e. to the cortico medullary (CM) epithelial cells and the follicle associated epithelium (FAE) supporting cells in guinea fowl and to the latter ones in the chicken. The expression of the K-1 antigen by these epithelial components may support their structural identity. After hatching, the K-1 molecule is restricted to the CM epithelial cells and/or FAE supporting cells, which suggests that the function of the embryonic epithelial bud is taken over by the CM epithelial cells. The K-1 positive CM epithelial cells form arches, which encompass blast-like cells. The possible relationship of the CM epithelial cells and blast-like cells, which may represent the precursors of bursal secretory dendritic cells is discussed. PMID- 11906728 TI - Detection of feline immunodeficiency virus RNA by two nucleic acid sequence based amplification (NASBA) formats. AB - Feline immunodeficiency virus (FIV) is an AIDS-inducing lentivirus that infects domestic cats worldwide. Because of its clinicopathologic similarities to human immunodeficiency virus type 1 (HIV-1) infection, the FIV/cat infection system is a valuable animal model for investigating comparative aspects of HIV-1 biology. An assay that detects quickly and efficiently FIV RNA in relatively small volume samples of feline blood or other body fluids would be of benefit in studies of viral transmission and antiviral interventions. Nucleic acid sequence based amplification (NASBA) technology is particularly suited for the detection of RNA in a variety of body fluids. In this report, the development of two rapid, sensitive and versatile NASBA formats is described for the detection of FIV gag RNA in plasma from infected cats. RNA detection by either format was unaffected by the presence of feline plasma. The limits of detection were at least 200 copies of input RNA for both formats. Results from seropositive and seronegative feline plasma samples were clearly distinguishable. These results demonstrate that NASBA provides a rapid and sensitive alternative to RT-PCR and culture isolation for detecting FIV RNA in infected feline plasma. PMID- 11906729 TI - Alphaherpesvirus antigen quantitation to optimize the diagnosis of herpes B virus infection. AB - Standardized, quantified virus antigen stocks are essential for dependable quality control of diagnostic assays. Five simple, rapid and economical direct enzyme linked immunoassays (dELISA) were developed to standardize and optimize antigen from five major cross-reacting alphaherpesviruses: herpes B virus, herpesvirus papio 2, langur monkey herpesvirus, herpes simplex virus-1 and herpes simplex virus-2. Each dELISA relied on pools of convalescent sera from rhesus monkeys, baboons, langurs and humans. Conjugates were prepared from purified IgG preparations, fractionated from the same sera and then labeled with peroxidase. Serum coated microplates could be stored at -70 degrees C for at least 1 year before use. The duration of the test was approximately 2.5 h if plates were prepared at an earlier time. Virus antigen titers could be determined from titration curves or from single dilutions using a standard curve. The sensitivity of detection was approximately 8x10(5) PFU/ml. This sensitivity sufficed for the determination of viral antigen mass in live or detergent treated virus stocks that usually contain at least 1x10(8) PFU/ml. The assays were valuable for quality assurance of diagnostic serological assays for herpes B virus and other alphaherpesviruses. PMID- 11906730 TI - A reverse-transcription competitive PCR assay based on chemiluminescence hybridization for detection and quantification of hepatitis C virus RNA. AB - A reverse-transcription competitive PCR (RT-cPCR) combined with chemiluminescence hybridization was designed for the detection and quantitative determination of serum hepatitis C virus (HCV) RNA. The concentration of HCV RNA was calculated based on an external standard curve that was generated by coamplification of internal competitor and target sequences in serial dilutions. The detection limit of the chemiluminescence RT-cPCR was 100 copies/ml (94 IU/ml). Meanwhile, the linear range for quantitation extended from 850 copies/ml (795 IU/ml) to 4.95x10(7) copies/ml. The performance of the current assay for measuring circulating HCV levels from 26 anti-HCV-antibody positive patients was compared with that of branched-chain DNA (bDNA) and nested RT-PCR assays. Eighteen patients had HCV RNA levels that exceeded the quantitation limit by the chemiluminescence RT-cPCR, but only 11 patients were quantitation-positive by the bDNA. A significant correlation of the quantitation values was found between the chemiluminescence RT-cPCR and the bDNA (R2=0.8391). Among the eight patients with HCV RNA titers below the quantitation limit, four remained positive by the chemiluminescence cRT-PCR, demonstrating the results in agreement with those using the nested RT-PCR. Furthermore, good linearity was revealed for the HCV genotypes 1b, 2a, 2b in 3-order magnitude diluted serum samples. In conclusion, the proposed chemiluminescence RT-cPCR method can detect quantitatively HCV RNA as accurately as the bDNA method and has sensitivity as high as nested RT-PCR. PMID- 11906731 TI - New tools for the construction of replication-competent adenoviral vectors with altered E1A regulation. AB - We have designed new vectors for the construction of recombinant adenoviruses containing the early region 1A (E1A) gene under the transcriptional control of heterologous promoters. The normal E1A regulatory elements have been replaced by a convenient set of unique restriction enzyme sites, allowing for introduction of gene regulatory cassettes with relative ease. Subsequent rescue generates recombinant conditionally replicating adenovirus in which the transcription of E1A is under alternative control. This allows potentially cell-type specific expression of E1A, restricting efficient virus replication to chosen target cells. It is shown that in several viruses rescued using these constructs, E1A expression is regulated by the heterologous promoters in the expected manner. Specifically, a virus with E1A under the control of the human Cytomegalovirus Immediate Early promoter produced constitutively high levels of E1A. A second virus, with E1A expression regulated by the glucocorticoid-responsive Mouse Mammary Tumor Virus promoter produced E1A expression in a dose-dependent manner upon dexamethasone treatment. Efficient growth of this second virus also required the presence of dexamethasone. PMID- 11906732 TI - Real-time quantitative analysis of polyoma BK viremia and viruria in renal allograft recipients. AB - Polyoma BK virus (BKV) remains dormant in the urinary tract and circulating leucocytes and becomes reactivated during immunosuppression. BK viruria is prevalent in renal allograft recipients and BK viremia may be related to nephropathy and allograft rejection. How BK viruria and viremia are related in renal allograft patients is undefined. In this study, BKV copies in paired urine and serum samples of renal allograft recipients were measured by a real time quantitative polymerase chain reaction (Q-PCR) to test the hypothesis that their quantitative relationship might help to delineate viral reactivation patterns in these tissues. Urine and plasma samples from 44 renal allograft recipients with stable graft function were collected during outpatient follow-up and the genome copies of BKV were determined by Q-PCR. All patients showed quantifiable viremia and two groups of patients were identified: one group of patients (n=35) showed low viral load (median: 270/ml, range: 108-1000/ml) and the other group (n=9) with high viral load (median: 5x10(4)/ml, range: 2x10(4)-6x10(4)/ml). The corresponding median levels of viruria were 2000 and 900 ml. BK viremia and viruria were not related quantitatively. BK viremia/viruria were also not related to age, immunosuppression, time and source of renal grafts and serum creatinine levels. The absence of a quantitative relationship between BK viremia and viruria may reflect independent BKV reactivation in different tissues during immunosuppression. PMID- 11906733 TI - Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to Junin virus in rodents. AB - Junin virus is the etiological agent of Argentine hemorrhagic fever, a serious rodent-borne disease. An enzyme-linked immunosorbent assay (ELISA) to detect Junin virus IgG antibodies in rodents was evaluated using sera from 27 Calomys musculinus and five Calomys laucha, inoculated experimentally with a live attenuated strain of this arenavirus. The test performance was compared against an indirect immunofluorescence assay (IFA). The ELISA had a sensitivity and specificity of 100% and a reproducibility of 87.9% for samples with titers above the selected cut-off value. IFA had lower sensitivity (53%) with the same specificity. The ELISA results were similar, whether carried out on whole blood or serum samples, thus eliminating the need for serum separation. A high correlation (K=0.86) between ELISA and IFA results was obtained from 1011 wild sigmodontine and murine rodents collected within and outside of the Argentine hemorrhagic fever endemic area. These results indicate that Junin virus IgG ELISA is the most suitable assay for detection of Junin virus antibodies in rodent samples. PMID- 11906734 TI - Overexpression and purification of the hepatitis B e antigen precursor. AB - Circumstantial evidence suggests that the secreted hepatitis B virus (HBV) e antigen (HBeAg) and/or its 22 kDa precursor (P22) have an essential role in the establishment of persistent infection. In order to identify cellular proteins that could interact with P22, large amounts of this protein are required to perform pull-down assays. A plasmid was constructed encoding a recombinant P22 with a Histidine-tag at its N-terminal extremity (P22r). The initial attempts to overexpress P22r in a conventional Escherichia coli strain failed, most likely due to the presence of rare AGA/AGG codon clusters in the 3' part of the gene. To overcome this difficulty, P22r was overexpressed in the Epicurian coli BL21 codonplus (DE3)-RIL strain, which possesses extra copies of the ArgU gene that encodes the tRNA(AGA/AGG). In this strain, P22r was overexpressed successfully and then purified in milligram quantities by metal affinity chromatography on Ni2+-chelated His-Bind resin. The purified recombinant protein P22r was able to interact with a cellular protein (P32), which had previously been shown to co immunoprecipitate with native P22, indicating that at least some of the P22r molecules were folded correctly. PMID- 11906735 TI - Cloning and molecular characterization of human high affinity antibody fragments against Hepatitis C virus NS3 helicase. AB - Hepatitis C Virus (HCV) non-structural protein 3 (NS3) helicase is essential for viral replication. Cloning of human antibody fragments for binding and inhibiting HCV helicase intracellularly (intracellular immunization) was attempted. A phage display system was employed to isolate human sFv fragments. A large phagemid library was cloned from patients infected with HCV. Phages expressing human sFv fragments with binding activity against NS3 were highly enriched during affinity selection. Selected sFv antibody fragments showed high affinity to HCV helicase. The variable domains of the cloned antibody fragments were sequenced and their germ-line origin was determined. K(D) values describing affinity of sFv to NS3 were measured by competition-EIA. Bacterially expressed recombinant human high affinity antibodies can be used for diagnostic and therapeutic purposes. Further experiments will select antibody fragments inhibiting NS3 helicase. Employing vectors for transduction of the encoding cDNA into infected cells might be a novel gene therapy strategy for intracellular immunization against chronic HCV infection. PMID- 11906736 TI - Enhanced detection of Theiler's virus RNA copy equivalents in the mouse central nervous system by real-time RT-PCR. AB - Infection of mice by low-neurovirulence Theiler's murine encephalomyelitis virus (TMEV), such as BeAn and DA viruses, provides a relevant experimental animal model for multiple sclerosis (MS). As a step toward determining the kinetics of a persistent central nervous system (CNS) infection that leads to chronic demyelination, we adapted a rapid, accurate and highly specific real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection and quantitation of BeAn virus RNA copy equivalents in mouse tissues. The assay enabled detection of as few as 20-30 copies of BeAn virus RNA per microg of total RNA from infected mouse tissues and results for spinal cord revealed the same high levels of BeAn RNA as detected by Northern hybridization during the first 4 months of the persistent infection, but also was able to detect virus RNA copies as late as 1 year post-infection. Real-time RT-PCR analysis of BeAn virus RNA copy equivalents in different parts of the CNS, analyses not possible by Northern hybridization, revealed the following cline of virus persistence: spinal cord>brainstem/cerebellum>cerebrospinal fluid (CSF)>cerebral hemispheres. Systemic organs, including heart, intestine and mesenteric lymph nodes of infected mice, showed no evidence of viral persistence at 4 months post infection. PMID- 11906738 TI - Can imperfections help to improve bioreactor performance? AB - Pilot-scale and larger bioreactors differ from small laboratory-scale reactors in terms of a greater occurrence of noise and incomplete mixing of the broth. Conventional control tries to induce good mixing and to filter out the noise as completely as possible. As such an 'ideal' operation is difficult to achieve, recent work has tried to exploit the non-ideal features to improve the performance. Using artificial neural networks, the degree of mixing, the extent of filtering of noise and the distribution of plasmid copy number (in a recombinant fermentation) can be controlled effectively on-line. This strategy generates better productivities than well-mixed noise-free operations, which suggests that deviations from ideal behaviour should be gainfully harnessed and not suppressed. PMID- 11906737 TI - Development and evaluation of a nucleic acid sequence based amplification (NASBA) protocol for the detection of enterovirus RNA in cerebrospinal fluid samples. AB - A nucleic acid sequence based (NASBA) assay for the generic detection of enterovirus RNA in cerebrospinal fluid (CSF) samples was developed and compared with an established reverse transcription/nested polymerase chain reaction (PCR) protocol. The sensitivity of NASBA followed by detection of amplicons with a biotinylated oligonucleotide probe was < or = ten copies of enterovirus RNA, indicating that enterovirus NASBA achieves a similar sensitivity as nested PCR. Moreover, NASBA detected a panel of 22 different serotypes of the species poliovirus, human enterovirus A, human enterovirus B and human enterovirus C completely. For evaluating NASBA as a diagnostic tool, 61 CSF samples of patients suffering from aseptic meningitis were tested in parallel with NASBA and nested PCR. NASBA detected enterovirus RNA in four CSF samples, two of these were also positive by nested PCR and two other CSF samples were positive only by nested PCR (in total six positive samples). All other 55 of 61 CSF samples were concordantly enterovirus negative by both methods. In conclusion, the more simple to handle 'one step' NASBA is as sensitive as nested PCR and may be used as an alternative method for the detection of enterovirus RNA in CSF samples. Enterovirus NASBA is a 'one step' RNA amplification procedure that is less prone to cross contamination compared to a three step nested PCR. PMID- 11906739 TI - Gene expression profiling diagnosis through DNA molecular computation. AB - Gene expression profiling is the characterization of cells based on the level of gene activity represented by concentrations of complementary DNA reverse transcribed from messenger RNA. The spectrum of cDNA concentrations, the expression profile, is determined using a DNA microarray. Although this approach is valuable for research, a simpler scheme that would give answers on a shorter time-scale for clinical applications is needed. An Adleman DNA self-assembly computer that would use cDNA as input might be ideal for clinical cell discrimination and a neural network architecture would be appropriate for making the necessary classifications. Preliminary experimental results suggest that expression profiling should be feasible using a DNA neural network that acts directly on cDNA. PMID- 11906740 TI - Applications of flow cytometry to ecotoxicity testing using microalgae. AB - Flow cytometry is a rapid method for the quantitative measurement of light scattering and fluorescent properties of cells. Although this technique has been widely applied to biomedical and environmental studies, its potential as a tool in ecotoxicological studies has not yet been fully exploited. This article describes the application of flow cytometry to the development of bioassays with marine and freshwater algae for assessing the bioavailability of contaminants in waters and sediments. PMID- 11906741 TI - Array for cancer prognostics. PMID- 11906742 TI - Finding the 'lost' genes. PMID- 11906744 TI - A novel trap for dynamic protein interactions in cytokine signaling. PMID- 11906745 TI - The worm in us - Caenorhabditis elegans as a model of human disease. PMID- 11906746 TI - Rapid monitoring of recombinant protein products: a comparison of current technologies. AB - Specific measurement of recombinant protein titer in a complex environment during industrial bioprocessing has traditionally relied on labor-intensive and time consuming immunoassays. In recent years, however, developments in analytical technology have resulted in improved methods for protein product monitoring during bioprocessing. The choice of product-monitoring technology for a particular bioprocess will depend on a variety of assay factors and instrument specific factors. In this article, we have compiled an overview of the advantages and disadvantages of the most commonly used technologies used: electrochemiluminescence, optical biosensors, rapid chromatography and nephelometry. The advantages of each technology for measuring both small and large recombinant therapeutic proteins are compared with a conventional enzyme linked immunosorbent assay (ELISA) technique. PMID- 11906747 TI - The legacy of nuclear risk and the founder effect in biotechnology organizations. AB - In the wake of the Chernobyl nuclear accident and a decline in the public trust of science, the founders of modern biotechnology recognized the strategic importance of risk assessment and regulatory affairs. In an effort to avoid the demonization that was attached to the nuclear industry, the pioneers of modern biotechnology delegated authority for regulatory negotiation and risk management to senior positions in the firm. At the same time, the Biotechnology Industry Organization was handed great latitude and trust with making public pronouncements on issues of bioethics and public policy. The way in which founders and leaders embed norms for negotiating regulation and responding to public perceptions has proved important in the maturation and acceptance of a biotechnology sector. PMID- 11906748 TI - Protein microarray technology. AB - Microarray technology allows the simultaneous analysis of thousands of parameters within a single experiment. Microspots of capture molecules are immobilized in rows and columns onto a solid support and exposed to samples containing the corresponding binding molecules. Readout systems based on fluorescence, chemiluminescence, mass spectrometry, radioactivity or electrochemistry can be used to detect complex formation within each microspot. Such miniaturized and parallelized binding assays can be highly sensitive, and the extraordinary power of the method is exemplified by array-based gene expression analysis. In these systems, arrays containing immobilized DNA probes are exposed to complementary targets and the degree of hybridization is measured. Recent developments in the field of protein microarrays show applications for enzyme-substrate, DNA-protein and different types of protein-protein interactions. Here, we discuss theoretical advantages and limitations of any miniaturized capture-molecule-ligand assay system and discusses how the use of protein microarrays will change diagnostic methods and genome and proteome research. PMID- 11906749 TI - Novel miniaturized systems in high-throughput screening. AB - High-throughput screening (HTS) using high-density microplates is the primary method for the discovery of novel lead candidate molecules. However, new strategies that eschew 2D microplate technology, including technologies that enable mass screening of targets against large combinatorial libraries, have the potential to greatly increase throughput and decrease unit cost. This review presents an overview of state-of-the-art microplate-based HTS technology and includes a discussion of emerging miniaturized systems for HTS. We focus on new methods of encoding combinatorial libraries that promise throughputs of as many as 100,000 compounds per second. PMID- 11906753 TI - Some questions to be raised about the hepatitis B vaccine. AB - The author consider some questions to be more carefully studied about the hepatitis B vaccine: route of injection (subcutaneous or intramuscular), amount of antigen (Ag), adjuvant (necessary or not), antigenic and molecular structure of HBsAg. What is important is to get high seroconversion rates, but not to get very high antibody titers (protective immunity is not dependent of antibody titers). Two types of vaccine could be used, and further studied. Why not use in healthy adults and children low dose vaccines (with 1.5-2.5 microg of HBsAg), possibly without adjuvant, and to be injected subcutaneously? And the 20 microg dose of vaccine could be used intramuscularly only for low- or non-responders, in high risk groups. PMID- 11906750 TI - Recombinant baculoviruses as mammalian cell gene-delivery vectors. AB - The baculovirus expression system has been used extensively for the expression of recombinant proteins in insect cells. Recently, recombinant baculovirus vectors engineered to contain mammalian cell-active promoter elements, have been used successfully for transient and stable gene delivery in a broad spectrum of primary and established mammalian cells. The application of modified baculoviruses for in vivo gene delivery has also been demonstrated. In contrast to other commonly used viral vectors, baculoviruses have the unique property of replicating in insect cells while being incapable of initiating a replication cycle and producing infectious virus in mammalian cells. The viruses can be readily manipulated, accommodate large insertions of foreign DNA, initiate little to no microscopically observable cytopathic effect in mammalian cells and have a good biosafety profile. These attributes will undoubtedly lead to the increased application and continued development of this system for efficient gene delivery into mammalian cells. Who said you can't teach an old dog new tricks? PMID- 11906754 TI - Effectiveness of hepatitis A vaccine in a former frequently affected community: 9 years' followup after the Monroe field trial of VAQTA. AB - The Kiryas Joel community in Monroe, N.Y. was the site of the first clinical trial which proved the protective efficacy of hepatitis A vaccine. The vaccine used was VAQTA J. Med. Virol (hepatitis A vaccine, inactivated). In the 9 years since the trial ended vaccination of infants reaching 2 years of age has continued along with monitoring for hepatitis A cases. The prevaccine pattern of frequent outbreaks has converted to a sustained pattern of no outbreaks, despite sporadic introduction of hepatitis A into the community in nonvaccinees. Community use of VAQTA in children 2 years of age and older has proven capable of providing long-term prevention of hepatitis A outbreaks in a formerly frequently affected community despite prolonged sporadic introduction of the virus. PMID- 11906755 TI - Simultaneous GeneGun immunisation with plasmids encoding antigen and GM-CSF: significant enhancement of murine antivenom IgG1 titres. AB - GeneGun DNA immunisation is a potent means of inducing antibody-dominant immune responses that we are exploiting to generate venom toxin-specific antibodies to improve the therapy of systemic envenoming by snakes. Here, we report that mice immunised with DNA encoding the carboxyl domain (JD9) of a haemorrhagic Zn metalloprotease (Jararhagin) in venom of the South American pit viper, Bothrops jararaca, and a plasmid expressing murine cytokine granulocyte/macrophage-colony stimulating factor (GM-CSF) raised significantly higher antigen-specific IgG1 titres than mice immunised with JD9 DNA alone. Serological responses to GeneGun JD9 DNA immunisation were shown to be dominated by IgG1, an IgG subclass associated with T lymphocyte helper 2 (Th2) immune responses. Further significant enhancement of JD9-specific IgG1 titres was achieved by increasing the number of immunisations. This report illustrates that DNA immunisation protocols to achieve high-titre, venom toxin-specific antibody production are well advanced and encourage the development of a DNA-based approach to antivenom production. PMID- 11906756 TI - Remarkably high antibody levels and protection against P. falciparum malaria in Aotus monkeys after a single immunisation of SPf66 encapsulated in PLGA microspheres. AB - Single dose immunisation is a major goal in vaccine design. The purpose of this study was the development of a single dose delivery system for the SPf66 malaria vaccine, based on this antigen's microencapsulation in PLGA microspheres by double emulsion method. Results indicate that a single immunisation in mice and monkeys with the SPf66 malaria vaccine, encapsulated in a mixture of two formulations of PLGA microspheres, induced a remarkably high and long-lasting immune response as assessed by ELISA and Western Blott. This immune response was associated with a good protective capacity in Aotus monkeys, after experimental challenge, indicating that antigen integrity lasted following the microencapsulation process. PLGA biodegradable microspheres thus serve as an effective delivery system for the design of a single dose immunisation vaccine, such as the SPf66 synthetic malaria vaccine. PMID- 11906757 TI - Distribution of pertussis antibodies among different age groups in Japan. AB - We examined the distribution of antibody levels against pertussis toxin (PT) and filamentous hemagglutinin (FHA) among a healthy Japanese population aged from 0 to 77 years. Levels of both antibodies in 1108 serum samples collected in 1994 from nine prefectures were assayed using polystyrene ball ELISA. The ratio of individuals positive (>or=10 ELISA U/ml) for anti-PT and anti-FHA antibodies at ages ranging from 0 to 3 years increased rapidly with the increase in the population vaccinated over three times with acellular pertussis (aP) vaccine. However, the ratio of those positive for anti-PT antibody tended to decrease until 6-8 years of age and to increase again from 9 to 19 years among the vaccinated population, although the ratio of individuals positive for anti-FHA antibody remained constant at 80-100% in children and adolescents over 3 years old. Moreover, positivity for anti-PT antibody was high (>or=50 ELISA U/ml) in some serum samples collected from adolescents and young adults, suggesting recent symptomatic or asymptomatic infection with circulating Bordetella pertussis. On the other hand, 50-60% of infants below 12 months of age was below the detection limit (1.0 ELISA U/ml) for anti-PT and anti-FHA antibodies, and most early infants were not vaccinated for pertussis. Since intermittent circulation of B. pertussis remains among the Japanese population, complete vaccination with aP vaccine for all infants should be highly recommended to prevent pertussis. PMID- 11906758 TI - Facilitation of antibody forming responses to viral vaccine antigens in young cats by recombinant baculovirus-expressed feline IFN-gamma. AB - We assessed the effect of recombinant feline IFN-gamma as vaccine adjuvant for in vivo antibody responses of young 3-month-old kittens to inactivated antigens of rabies and calicivirus, both natural pathogens for cats. When compared to responses following immunization with antigen alone co-administration of baculovirus-expressed cat IFN-gamma significantly enhanced serum antibody titers to both viral antigens; to levels comparable with responses evoked by commonly known saponin and alum adjuvants. Adjuvanticity by feline IFN-gamma was dose dependent and all doses tested were well tolerated. We conclude that, when further optimized for in vivo delivery, feline IFN-gamma may represent a safe and efficient natural vaccine adjuvant for certain antigens in cats. PMID- 11906759 TI - Injectable silicone implants as vaccine delivery vehicles. AB - Injectable silicone implants were assessed as vaccine delivery vehicles in sheep, using either the model antigen avidin or Clostridium tetani and Clostridium novyi toxoids. Two types of implant were compared, the matrix type, that has been shown to deliver antigen in vitro in a first-order profile over approximately 1 month, and the covered rod type, that delivers antigen for several months in a zero order profile. The implants were prepared using lyophilized antigen and adjuvant (in this case, recombinant ovine interleukin-1beta; rovIL-1beta) and manufactured in the absence of extremes of temperature or pH or the use of organic solvents. Use of the matrix type implant was capable of inducing antibody responses equivalent to those induced by conventional vaccination with aluminium hydroxide adjuvant ("alum"). The use of the covered rod implants, that release very low levels of antigen over a long period, induced responses that were markedly enhanced over the alum control groups. The covered rod implant also favoured production of both IgG1 and IgG2 isotypes in contrast to responses of matrix vaccinated sheep and conventionally vaccinated control sheep in which IgG1 predominated. Prolonged duration of the antibody response was also observed following vaccination with covered rod implants. Dose-response analysis using the matrix implant demonstrated a trend towards improved responses for lower antigen doses. Clostridial vaccination of sheep showed that protective antibody titres up to 4-fold higher than for alum-adjuvanted groups could be induced by administering the antigen in the covered rod implant. Responses elicited by all implant groups were dependent on the inclusion of adjuvant into the implant formulation. PMID- 11906760 TI - The level of protection against rotavirus shedding in mice following immunization with a chimeric VP6 protein is dependent on the route and the coadministered adjuvant. AB - Intranasal (i.n.) immunization of BALB/c mice with chimeric murine rotavirus EDIM (epizootic diarrhea of infant mice) VP6 and attenuated E. coli heat-labile toxin (LT), LT(R192G), stimulated >99% protection against rotavirus shedding after EDIM challenge. Here, we evaluated other potential adjuvants with chimeric VP6 administered by two mucosal routes: i.n. and oral. Besides LT(R192G), the adjuvants examined included Adjumer, CpG oligodeoxynucleotides (CpG ODN), chimeric A1 subunit of cholera toxin (CTA1)-DD, and QS-21. All except QS-21 significantly (P<0.05) increased VP6-specific serum IgG responses after i.n. immunization, but none significantly increased these responses when administered orally. The i.n. delivery of chimeric VP6 alone induced both rotavirus IgG1 and IgG2a whose relative titers suggested a skewed Th2-like response. Inclusion of Adjumer greatly increased Th2-like responses, while CpG ODN shifted the response to a less Th2-like response. The adjuvants CTA1-DD, LT(R192G), QS-21 had no significant effect on ratios of IgG1/IgG2a titers. Following EDIM challenge of mice immunized i.n. with chimeric VP6 and either LT(R192G), CTA1-DD, Adjumer or CpG ODN, shedding was reduced >99, 95, 80, 74, respectively, relative to that found in unimmunized mice (P<0.05). QS-21 induced less protection (43%, not significant (N.S.)) while immunization with chimeric VP6 alone reduced shedding by only 16% (N.S.). Oral immunization with chimeric VP6 and all selected adjuvants except QS-21 was less effective than after i.n. immunization, with protection levels of 94 (P<0.05), 71 (P<0.05), 55, 35 and 28% for LT(R192G), QS 21, CpG ODN, CTA1-DD, and Adjumer, respectively, while immunization with chimeric VP6 alone gave no protection. Thus, different adjuvants induced different degrees of protection and oral immunization was generally less effective then the i.n. route. PMID- 11906761 TI - Systemic and intestinal antibody secreting cell responses and protection in gnotobiotic pigs immunized orally with attenuated Wa human rotavirus and Wa 2/6 rotavirus-like-particles associated with immunostimulating complexes. AB - The undesirable side effects and variable efficacy of some oral live rotavirus vaccines in infants have necessitated alternative vaccine approaches. We evaluated a recombinant RFVP2/WaVP6 rotavirus-like-particle (2/6VLP) oral vaccine, using an immunostimulating complex (ISCOM) matrix as adjuvant, in a gnotobiotic (Gn) pig model of human rotavirus (HRV) disease. The 2/6VLPs adhered to the ISCOM-matrix (2/6VLP-ISCOM ) and were antigenic, but they failed to induce protection. However, when combined with attenuated (Att) HRV for oral priming, the 2/6VLP-ISCOM vaccine was effective as a booster and induced partial protection against virulent Wa HRV. The 250 microg 2/6VLP dose was more effective than 100 microg. The highest mean numbers of IgA antibody secreting cells evaluated by ELISPOT in intestinal lymphoid tissues were in pigs receiving AttHRV+2/6VLP-ISCOM or three doses of AttHRV and were associated with the highest protection rates. PMID- 11906762 TI - The outer membrane proteins UspA1 and UspA2 of Moraxella catarrhalis are highly conserved in nasopharyngeal isolates from young children. AB - UspA1 and UspA2 of Moraxella catarrhalis are vaccine candidates. The aims of this study were to determine: (1) the frequencies of occurrence and (2) the degrees of conservation of two surface-exposed epitopes of the uspA1 and uspA2 genes and their respective gene products in 108 nasopharyngeal isolates from young children. The uspA1 and uspA2 genes were detected in 107 (99%) and 108 (100%) isolates, respectively. Twenty-three of 108 uspA2 genes (21%) were identified as the variant gene uspA2H. One-hundred and five isolates (97%) expressed the mAb17C7-reactive epitope shared by UspA1 and UspA2, and 103 isolates (95%) reacted with the UspA1-specific mAb24B5. The only isolate which lacked a uspA1 gene demonstrated reduced adherence to HEp-2 cells and complement sensitivity. The data indicate that both uspA genes and the expression of at least two surface exposed epitopes are virtually ubiquitous in isolates from a population at risk for otitis media. A vaccine capable of inducing a bactericidal immune response against the mAb17C7- and/or mAb24B5-reactive epitopes appears promising. PMID- 11906763 TI - Induction of specific Th1 responses and suppression of IgE antibody formation by vaccination with plasmid DNA encoding Der f 11. AB - DNA vaccines encoding low-molecular-weight allergens have been used to prevent IgE responses. A high-molecular-weight mite allergen Der f 11 that was hardly to be purified for immunotherapy was used to develop a DNA vaccine here. Vaccination of mice with plasmid DNA encoding Df11 (pDf11) induced Th1 responses characterized by IgG2a responses and spleen cell secretion of IFN-gamma. In contrast, sensitization with recombinant Der f 11 (rDf11) and alum induced Th2 responses characterized by IgE responses and spleen cell secretion of IL-4 and IL 5. Vaccination with pDf11 prevented the induction of IgE responses. Moreover, it could inhibit on-going IgE responses. The debate whether CD4+ or CD8+ T cells were the regulatory cells to inhibit IgE responses by DNA vaccination was also examined. First, sensitization of pDf11-vaccinated mice after depletion of CD8+ T cells still showed suppression of IgE responses. Secondly, adoptive transfer of either CD4- or CD8-depleted spleen cells from pDf11-vaccinated mice suppressed IgE responses. In conclusion, this is the first report to confirm the therapeutic effect of a DNA vaccine encoding a strong allergen on specific IgE responses. Both CD4+ and CD8+ T cells are crucial for the immunomodulation of IgE responses by pDf11. PMID- 11906764 TI - Comparison of the immune responses induced by local immunizations with recombinant Lactobacillus plantarum producing tetanus toxin fragment C in different cellular locations. AB - Lactobacillus plantarum NCIMB8826 was selected as a bacterial carrier for the development of live mucosal vaccines. This strain was reported to display interesting pharmaco-kinetic properties when fed to human volunteers and is also able to persist in the mouse intestine. The non-toxic C fragment of tetanus toxin (TTFC) was used as a model antigen. Recombinant strains producing TTFC in three cellular locations, intracellular, secreted or cell-surface exposed were compared to each other by immunizing mice through the subcutaneous, intranasal and intragastric routes. The three types of constructs were able to induce strong specific immune responses against TTFC by all routes tested. While cell-surface presentation required lower antigen doses to be immunogenic, the highest IgG serum antibody titers were obtained with the strain producing large amounts of TTFC in the cytoplasm. PMID- 11906765 TI - Estimating the potential health gain and cost consequences of introducing a pre school DTPa pertussis booster into the UK child vaccination schedule. AB - This work estimates the health and cost impacts of a pre-school booster vaccination for Bordetella pertussis, when added to existing UK primary vaccination. A transition state simulation model of pertussis infection in a closed population was constructed comprising of susceptible, infected and immune population sub-groups, across eight age bands. Epidemiological, service use and cost data were sourced from routine statistics, published literature and clinician estimates. The introduction of a pre-school booster is predicted to reduce the number of hospitalisations by approximately 1400 and pertussis cases suffered by up to 28,000 at a net investment of under 13 million pounds over a 5 year period. PMID- 11906766 TI - DNA vaccination against feline calicivirus infection using a plasmid encoding the mature capsid protein. AB - Feline calicivirus (FCV), a member of the diverse family Caliciviridae, is a respiratory and oral pathogen of cats. Although conventional FCV vaccines are available, there are some safety and efficacy problems associated with their use. The potential of DNA vaccination against FCV infection was therefore explored. Four cats were inoculated intramuscularly with three 100 microg doses, 2 weeks apart, with a plasmid (pF9VAC) containing the mature capsid protein gene of FCV strain F9. Four control cats received the same plasmid lacking the FCV gene insert. All eight cats showed clinical signs following heterologous challenge with FCV strain LS027. However, rectal temperatures and general clinical sign scores were significantly lower in vaccinates compared to controls, and there was a marked difference in ulcer distribution between the two groups. Although no serological responses were detected in either group prior to challenge, post challenge titres in the vaccinated group were generally higher. The results indicate that partial protection against a calicivirus is possible by DNA vaccination but that other approaches to enhance efficacy such as the use of cytokine genes or prime-boost protocols may also be required. PMID- 11906767 TI - Evaluation of vaccines for atrophic rhinitis--a comparison of three challenge models. AB - We compared three challenge models for the assessment of atrophic rhinitis (AR) vaccines: combined infection with Bordetella bronchiseptica (Bb) and Pasteurella multocida (Pm); application of acetic acid (AA) to the nasal mucosa followed by Pm infection; and Bb infection alone. Two vaccines were tested using standardized criteria, notably nasal lesion scores. The vaccines provided different levels of protection in the Bb and the AA/Pm challenges, but were similar in the combined (Bb/Pm) challenge. It is clear that the AA/Pm model shows the protective value of only the Pm component, whereas the single Bb challenge reflects the protective value merely of the Bb component of a combined vaccine. These results suggest that the best assessment of protection is provided if the two specific challenges are performed separately. PMID- 11906768 TI - Comparative efficacy, safety and immunogenicity of Hepavax-Gene and Engerix-B, recombinant hepatitis B vaccines, in infants born to HBsAg and HBeAg positive mothers in Vietnam: an assessment at 2 years. AB - In a randomized, controlled trial, 105 healthy full-term infants born to HBsAg and HBeAg positive mothers received three doses (at 0, 1 and 6 months) of either a new recombinant hepatitis B vaccine (Hepavax-Gene) or Engerix-B. Both groups were also given hepatitis B specific Hepa-big immunoglobulin (HBIG) within 24h of birth. Levels of antibodies to hepatitis B surface antigen (anti-HBs) were assessed on days 30, 60, 210, 360, and 2 years post-vaccination. Efficacy and immunogenicity and safety of the two vaccines were not significantly different; both vaccines achieved >94% seroprotection within 360 days. At 2 years, only one subject (1.9%) in the Hepavax-Gene group and two subjects (3.9%) in the Engerix-B group were HBsAg positive. No serious adverse events (AEs) were observed in either group. PMID- 11906769 TI - Lipopeptide stimulation of MHC class I-restricted memory cytotoxic T lymphocytes from equine infectious anemia virus-infected horses. AB - The immunogenicity of equine infectious anemia virus (EIAV) Gag and Env equine leukocyte alloantigen (ELA)-A5.1, -A9, and -A1 restricted cytotoxic T lymphocyte (CTL) epitopes synthesized on multiple antigenic peptide (MAP) system coupled to tripalmitoyl-S-glycerylcysteine (P3C) was evaluated in vitro. P3C-MAP-peptide stimulated peripheral blood mononuclear cells (PBMCs) from horses, chronically infected with EIAV, had memory CTL (CTLm) similar to that of PBMCs stimulated with either the minimal CTL epitopes, longer peptides containing the CTL epitopes, or EIAV. The stimulated CTL lysed EIAV-infected target cells and the percent specific lysis was dependent on the dose of P3C-MAP-peptide used to stimulate PBMCs in vitro and was peptide specific and ELA-A restricted. The results suggest that these P3C-MAP-peptides can be used as immunogens to stimulate primary immune responses in vivo. PMID- 11906770 TI - Brucella abortus vaccine strain RB51 produces low levels of M-like O-antigen. AB - Brucella abortus RB51 is a rough (R) stable vaccine strain used in cattle and is believed to be devoid of O-side chain. We analyzed by use of a panel of monoclonal antibodies (MAbs) directed against seven previously defined O polysaccharide (O-PS) epitopes the O-chain expression in strain RB51. Two MAbs specific for the C/Y (A=M) and C (M>A) epitopes showed low bindings in ELISA to strain RB51. O-chain expression was further confirmed by Western blot after SDS PAGE of strain RB51. In particular, the MAb of C (M>A) specificity, showing preferential binding to M-dominant smooth (S) Brucella strains, revealed in strain RB51 a typical smooth-lipopolysaccharide (S-LPS) pattern which resembled that of M-dominant S-LPS. Thus, the results clearly show that strain RB51 produces low levels of M-like O-antigen. PMID- 11906772 TI - A meta-analysis of effectiveness of influenza vaccine in persons aged 65 years and over living in the community. AB - AIM: To estimate the effectiveness of inactivated influenza vaccine in persons aged 65 years and over living in the community. SCOPE: A meta-analysis of studies selected using predetermined criteria without language restriction. CONCLUSION: Influenza vaccine was effective in reducing influenza-like illness by 35% (95% confidence interval (CI) 19-47%), hospitalization for pneumonia and influenza by 33% (CI 27-38%), mortality following hospitalization for pneumonia and influenza by 47% (CI 25-62%); and mortality from all causes by 50% (CI 45-56%). PMID- 11906771 TI - Immune response to synthetic peptides of dengue prM protein. AB - The immunological activities of five synthetic peptides of the prM protein of dengue-2 (DEN-2) virus containing B cell epitopes were evaluated in BALB/c mice. Two peptides elicited neutralizing antibodies against all four DEN serotypes. Virus-specific proliferative responses were demonstrated in mice immunized with four of the five peptides, demonstrating the presence of T cell epitopes. Mice immunized with three of the five peptides conjugated with bovine albumin showed statistically significant levels (P<0.05) of protection when challenged with DEN 2 virus. These results could constitute the basis for the establishment of the role of DEN virus pre and M antigens in the development of anti-flaviviral vaccines. PMID- 11906773 TI - Maternal immunization with pneumococcal polysaccharide vaccine in the highlands of Papua New Guinea. AB - In Tari, Southern Highlands Province (SHP), Papua New Guinea (PNG), pneumococcal polysaccharide (Pnc PS) vaccine was offered to women at 28-38 weeks gestation. Blood samples were collected for measurement of pneumococcal antibody titres prior to immunization, from mother and cord at delivery and from their children at ages 1-3 and 4-6 months; samples were also collected in a subset of children before and 1 month after Pnc PS vaccine was given at age 8-9 months. Serum was collected from unimmunized women and their children at delivery and from children of unimmunized women at the same ages in infancy. There were no differences in neonatal or post-neonatal mortality rates or congenital abnormalities in the children of 235 immunized and 202 unimmunized women. There was a significant increase in antibody titres to pneumococcal serotypes 5, 14 and 23F in immunized women but not for serotype 7F. Geometric mean titres (GMTs) of antibodies for serotypes 5 and 23F were significantly higher in children of immunized women than in the unimmunized group up to age 2 months and for serotype 14 significantly higher to age 4 months. Maternal immunization did not significantly affect the children's capacity to make antibody responses to immunization with Pnc PS vaccine in infancy. The findings of this study and those in several other developing countries provide support for the concept of Pnc PS maternal immunization and justify the planning of large-scale efficacy trials. PMID- 11906774 TI - The recombinant chimeric human parainfluenza virus type 1 vaccine candidate, rHPIV3-1cp45, is attenuated, immunogenic, and protective in African green monkeys. AB - A recombinant live-attenuated chimeric human parainfluenza virus type 1 (HPIV1) candidate vaccine was previously generated by replacing the fusion (F) and hemagglutinin-neuraminidase (HN) glycoprotein open reading frames (ORFs) of the HPIV3 candidate vaccine, rHPIV3cp45, with those of wild-type HPIV1. Previously, this recombinant chimeric virus, designated rHPIV3-1cp45, exhibited a greater level of the temperature sensitivity of replication in vitro and a greater level of attenuation of replication in the respiratory tract of immunized hamsters when compared to its HPIV3cp45 parent virus. In the present study, rHPIV3-1cp45 was evaluated for its level of attenuation and efficacy in African green monkeys (Cercopithecus aethiops), a primate in which both HPIV1 and HPIV3 wild-type viruses replicate efficiently. The rHPIV3-1cp45 candidate vaccine was as restricted in replication in the upper and lower respiratory tract as its thoroughly characterized rHPIV3cp45 parent indicating that the attenuating mutations present in the rHPIV3cp45 backbone specified an appropriate level of attenuation of rHPIV3-1cp45 for primates. The level to which rHPIV3-1cp45 replicated in the respiratory tract of African green monkeys was also sufficient to induce a strong immune response to HPIV1 and provided protection against challenge with wild-type HPIV1. These results provide a basis for further evaluation of this HPIV1 candidate vaccine in humans. PMID- 11906775 TI - Delayed-type hypersensitivity in volunteers immunized with a synthetic multi antigen peptide vaccine (PfCS-MAP1NYU) against Plasmodium falciparum sporozoites. AB - During the testing of the safety and immunogenicity of an adjuvanted, synthetic Plasmodium falciparum CS multiple antigen peptide (MAP) vaccine, we investigated the potential for using cutaneous delayed-type hypersensitivity (DTH) reactions as a correlate of immune response. We evaluated 27 of our volunteers for DTH reactions to intradermal inoculation (0.02 ml) of several concentrations of the MAP vaccine and adjuvant control solutions. Induration was measured 2 days after skin tests were applied. Nine of 14 vaccinees (64%) with serum, high-titered anti MAP antibody developed positive DTH (>or=5mm induration), that first appeared by 29 days after immunization and persisted for at least 3-6 months after 1-2 more immunizations. In contrast, DTH responses were negative in eight of eight vaccinees with no or low antibody titers, and in five of five non-immunized volunteers. Biopsies of positive DTH skin test sites were histologically compatible with a DTH reaction. We conclude that the presence of T cell functional activity reflected by a positive DTH skin test response to the MAP antigen serves as another marker for vaccine immunogenicity. PMID- 11906776 TI - Insertion of interleukin-2 (IL-2) and interleukin-12 (IL-12) genes into vaccinia virus results in effective anti-tumor responses without toxicity. AB - Identification of novel tumor-associated antigens (TAA) capable of eliciting T cell responses has renewed interest in the development of anti-tumor vaccines. The insertion of genes encoding specific TAA into a vaccinia virus (rVV) is one approach to vaccination since large amounts of foreign DNA can be stably integrated into the poxvirus genome. Recent reports have documented an increased therapeutic effectiveness of poxvirus-based vaccines when additional treatment with cytokines, such as interleukin-2 (IL-2) or interleukin-12 (IL-12) were used, but the combination of these cytokines as adjuvants for a rVV encoding TAA have not been previously reported. The combination of IL-2 and IL-12 at single regimen systemic doses was toxic and sometimes fatal, manifesting largely as segmental epithelial apoptosis of the large bowel. To explore the local delivery of both cytokines to the site of vaccination, the genes encoding IL-2 and IL-12 were inserted into vaccinia virus along with a model tumor antigen gene. This construct contained five heterologous genes: LacZ (the model antigen), gpt (reporter gene), IL-2, and the two IL-12 subunit genes (p35 and p40). Treatment with this recombinant virus resulted in a reduced number of pulmonary metastases, improved survival, and minimal toxicity in a murine tumor model. The use of vaccinia virus for the insertion of other heterologous gene combinations may provide a powerful and less toxic approach for novel vaccination strategies in the treatment and prevention of cancer. PMID- 11906777 TI - Enhanced immunogenicity of a hepatitis B virus peptide vaccine using oligosaccharide ester derivative microparticles. AB - Controlled release microspheres can overcome many of the disadvantages of multiple vaccine delivery such as rate of uptake and cost of administration. Proteins and peptides are difficult to administer using conventional polymers owing to protein degradation, premature release and stability. Here we report the successful development of room temperature stable, controlled release formulations using oligosaccharide ester derivatives (OEDs) of trehalose and a synthetic peptide analogue of hepatitis B surface antigen. Employing a range of different OED preparations, we have optimised the immunogenicity of the peptide formulation such that mice injected with a single preparation of microspheres consisting of trehalose octaacetate (TR101; Group G) produce high titre anti hepatitis B (anti-HBs) surface antigen antibodies. The kinetics of the immune response could be manipulated with different peptide/OED formulations and correlated with the OED composition of the microspheres. Our data demonstrate the considerable potential of OED microspheres as novel delivery systems for vaccines. The ability to induce strong immune responses, without the requirement for multiple doses or cold-chain storage, could radically improve vaccination programmes in developing countries. PMID- 11906778 TI - Oxidised mannan-listeriolysin O conjugates induce Th1/Th2 cytokine responses after intranasal immunisation. AB - Clearance of infectious organisms does not always require polarised Th1 or Th2 responses and it may be advantageous for both Th1 and Th2 responses to be elicited for effective protection against an invading pathogen. It was the aim of this study to investigate oxidised mannan as a possible Th1/Th2 adjuvant. Oxidised mannan was conjugated to two candidate antigens and delivered intranasally to mice. Immunisation with the oxidised conjugate resulted in significant antigen specific proliferative responses, IL-2, IFN-gamma and IL-4 production when compared to unconjugated controls. PMID- 11906779 TI - DTPw-HB and Hib primary and booster vaccination: combined versus separate administration to Latin American children. AB - This multicentre study was designed to establish the reactogenicity and immunogenicity profiles of primary and booster vaccination with diphtheria, tetanus, and pertussis whole-cell-hepatitis B/Haemophilus influenzae type-b (DTPw HB/Hib) administered as either a syringe mix or as separate injections in 400 Latin American children. Both vaccine regimens were equally well tolerated and elicited post-primary excellent seropositivity rates at or close to 100% for all five component antigens. With regard to HB, 100% of subjects in the combined vaccination group, and 98.8% subjects in the separate injection vaccination group reached seroprotective antibody concentrations (>or=10 mIU/ml) 1 month after the primary vaccination course. Equally high anti-PRP antibody concentrations were reached 1 month after vaccination, with 100% of seroprotected subjects in the combined vaccination group (antibody concentrations >or=0.15 microg/ml), against 99.4% in the separate injection vaccination group. Seroprotective anti-HBs and anti-PRP antibody concentration levels persisted approximately 1 year after the primary vaccination course, just prior to booster vaccination. Finally, a significant increase of all antibody concentrations could be observed after the booster vaccination, since all but one subject in the separate injection vaccination group had protective levels of anti-HBs and anti-PRP antibodies 1 month after the booster dose. These results suggest that the combination of DTPw HB and Hib vaccines provides an effective means for increasing vaccine coverage in childhood vaccination programmes. PMID- 11906780 TI - Impaired release of beta-endorphin in response to serotonin in a rat model of depression. AB - Involvement of both the serotonergic and the endogenous opioid systems in the onset of depressive behavior has been suggested. Previously we showed that serotonin (5-hydroxytryptamine) facilitates beta-endorphin release in the nucleus accumbens (NAcc). Herein, the microdialysis method was used to assess in vivo the effects of serotonin on beta-endorphin release in a rat model of depressive behavior (the Flinders sensitive line, FSL), before and after antidepressant treatment. The basal extracellular level of beta-endorphin in the NAcc of FSL rats did not differ significantly from that in control rats. However, serotonin induced beta-endorphin release was impaired in FSL rats. Chronic treatment (18 days) with desipramine or paroxetine did not significantly affect the extracellular levels of beta-endorphin in the NAcc of either the FSL or control rats. However, the chronic antidepressant treatment did normalize the serotonin induced release of beta-endorphin in FSL rats, as well as their behavioral manifestation of depressive behavior. Our results show that depressive behavior may relate to an impaired effect of serotonin on beta-endorphin release in the NAcc in a rat model of depression, and suggest a possible new mode of action of antidepressant drugs. PMID- 11906781 TI - Properties and origin of spikelets in thalamocortical neurones in vitro. AB - Spikelets, or fast prepotentials as they are frequently referred to, are a common feature of the electrophysiology of central neurones and are invariably correlated with the presence of electrotonic coupling via gap junctions. Here we report that in the presence of the metabotropic glutamate receptor agonists, trans-ACPD or DHPG, thalamocortical neurones of the cat dorsal lateral geniculate nucleus maintained in vitro exhibit stereotypical spikelets that possess similar properties to those described in other brain areas. These spikelets were routinely observed in the presence of antagonists of fast chemical synaptic transmission, were resistant to the application of a variety of voltage-dependent Ca(2+) channel blockers but were abolished by tetrodotoxin. In addition, spikelets were reversibly blocked by the putative gap junction blocker carbenoxolone and were nearly always accompanied by dye-coupling. These results indicate that thalamocortical neurones may be electrotonically coupled via gap junctions with spikelets representing attenuated action potentials from adjoining cells. We suggest that the presence of electrotonic communication between thalamocortical neurones would have major implications for the understanding of both physiological (Steriade et al., 1993; Sillito et al., 1994; Alonso et al., 1996; Neuenschwander and Singer, 1996; Weliky and Katz, 1999) and pathological (Steriade and Contreras, 1995; Pinault et al., 1998) synchronised electrical activity in the thalamus. PMID- 11906782 TI - Distribution and synaptic localisation of the metabotropic glutamate receptor 4 (mGluR4) in the rodent CNS. AB - Group III metabotropic glutamate receptors (mGluRs) are selectively activated by L-2-amino-4-phosphonobutyrate (L-AP4), which produces depression of synaptic transmission. The relative contribution of different group III mGluRs to the effects of L-AP4 remains to be clarified. Here, we assessed the distribution of mGluR4 in the rat and mouse brain using affinity-purified antibodies raised against its entire C-terminal domain. The antibodies reacted specifically with mGluR4 and not with other mGluRs in transfected COS 7 cells. No immunoreactivity was detected in brains of mice with gene-targeted deletion of mGluR4. Pre embedding immunocytochemistry for light and electron microscopy showed the most intense labelling in the cerebellar cortex, basal ganglia, the sensory relay nuclei of the thalamus, and some hippocampal areas. Immunolabelling was most intense in presynaptic active zones. In the basal ganglia, both the direct and indirect striatal output pathways showed immunolabelled terminals forming mostly type II synapses on dendritic shafts. The localisation of mGluR4 on GABAergic terminals of striatal projection neurones suggests a role as a presynaptic heteroreceptor. In the cerebellar cortex and hippocampus, mGluR4 was also localised in terminals establishing type I synapses, where it probably operates as an autoreceptor. In the hippocampus, mGluR4 labelling was prominent in the dentate molecular layer and CA1-3 strata lacunosum moleculare and oriens. Somatodendritic profiles of some stratum oriens/alveus interneurones were richly decorated with mGluR4-labelled axon terminals making either type I or II synapses. This differential localisation suggests a regulation of synaptic transmission via a target cell-dependent synaptic segregation of mGluR4. Our results demonstrate that, like other group III mGluRs, presynaptic mGluR4 is highly enriched in the active zone of boutons innervating specific classes of neurones. In addition, the question of alternatively spliced mGluR4 isoforms is discussed. PMID- 11906783 TI - Differences in expression, actions and cocaine regulation of two isoforms for the brain transcriptional regulator NAC1. AB - BTB/POZ proteins can influence the cell cycle and contribute to oncogenesis. Many family members are present in the mammalian CNS. Previous work demonstrated elevated NAC1 mRNA levels in the rat nucleus accumbens in response to cocaine. NAC1 acts like other BTB/POZ proteins that regulate transcription but is unusual because of the absence of identifiable DNA binding domains. cDNAs were isolated encoding two NAC1 isoforms differing by only 27 amino acids (the longer isoform contains 514 amino acids). The mRNAs for both isoforms were simultaneously expressed throughout the rat brain and peripheral tissues. Semi-quantitative reverse transcription-polymerase chain reaction analysis revealed that the mRNA of the longer isoform was more abundant than the mRNA of the shorter isoform. Western blot analysis demonstrated a similar unequal distribution between the isoforms in the CNS. The longer isoform was the more abundant of the two NAC1 proteins and the ratio between them differed throughout the rat brain. The shorter isoform was not detected in most of the examined peripheral tissues, suggesting differences from the CNS in post-transcriptional processing. Both isoforms repressed transcription in H293T cells using a Gal4-luciferase reporter system. However, the shorter isoform did not repress transcription as effectively as the longer isoform. Transfection of different ratios for both isoforms, in order to replicate the relative amounts observed throughout the CNS, supported an interaction between the isoforms. The net effect on transcriptional repression was determined by the ratio of the two NAC1 isoforms. Each isoform exhibited the subnuclear localization that is characteristic of many BTB/POZ proteins. A rapid and transient increase in the level of the shorter isoform occurred in the nucleus accumbens 2 h following a single i.p. cocaine injection. We conclude that the two isoforms of NAC1 may differentially affect neuronal functions, including the regulation of cocaine-induced locomotion. PMID- 11906784 TI - Rapid eye movement sleep deprivation modifies expression of long-term potentiation in visual cortex of immature rats. AB - During rapid eye movement (REM) sleep, activity of non-retinal origin is propagated into central visual-system pathways in a manner similar, in pattern and intensity, to central visual-system activity that is exogenously generated in waking. It has been hypothesized that REM sleep, which is more abundantly represented early in life than later, functions to provide adjunct 'afferent' input for shaping synaptic connectivity during brain maturation. Here we present data that support this proposal. Recent studies have described a developmentally regulated form of in vitro long-term potentiation (LTP) in the visual cortex that is experience- and age-dependent. In immature rats, suppression of retinal activation of the visual system by removal of visual experience (dark rearing) extends the age when the developmentally regulated form of LTP can be produced. This study tests whether suppression of REM-state activation of the visual system also lengthens the developmental period in which this specific form of LTP can be elicited. Young rats were deprived of REM sleep by the multiple-small-platforms over-water method during the typically latest week for induction of such LTP in slices of visual cortex. After this week, we could still induce LTP in slices from nearly all the REM-sleep-deprived rats (8/9) but not from age-matched rats that had not lost REM sleep (0/5). The control rats had been housed on large platforms that allow the animals to obtain REM sleep. Only body weights and the concentration of thyrotrophin-releasing hormone in the hypothalamus distinguished home-caged, normal-sleeping controls from both groups of platform animals. On all measures, stress levels were not dissimilar in the two platforms groups. After 7 days of behavioral suppression of REM sleep in immature rats, and consequent reduction of the intense, extra-retinal activity endogenously generated during this sleep state, we found that the period was extended in which developmentally regulated synaptic plasticity (LTP) could be elicited in slices of visual neocortex. These studies support the role of REM sleep and its associated neuronal activity in brain maturation. PMID- 11906785 TI - Zinc-rich neurones in the rat visual cortex give rise to two laminar segregated systems of connections. AB - Zinc-rich synaptic boutons in the neocortex arise from the neocortex itself. However, the precise organisation of these circuits is not known. Therefore, the laminar and areal pattern of zinc-rich cortico-cortical connections between visual areas was studied by retrograde tracing using intracerebral injections of sodium selenite. This tracer was injected in supragranular and infragranular layers in various cortical visual areas in order to precipitate zinc in the synaptic boutons, which was retrogradely transported to neuronal somata. Supragranular injections led to retrogradely labelled neurones in layer II-III, ipsilaterally and contralaterally. Neurones often appeared in groups or clusters. Infragranular injections labelled neurones in layers II-III, VI and, to a lesser extent, in layer V, both ipsilaterally and contralaterally. Neurones in layer VI formed a wide continuous band. Concerning the connections between visual (=occipital) areas, injections in occipital area 2, lateral part (Oc2L), rendered the largest number of retrogradely labelled neurones, which were located in occipital area 1 (Oc1), occipital area 2, medial part (Oc2M) and outside the visual cortex. Callosal zinc-rich projections were dense in the homotopic area but sparse in Oc1 and temporal cortex. Injections in Oc1 rendered moderate numbers of labelled neurones in occipital areas, in both hemispheres. Injections in Oc2M labelled moderate numbers of neurones in occipital areas in both hemispheres and in the frontal and cingulate cortices. These results indicate that zinc-rich cortico-cortical connections are organised into two segregated systems arising from either supragranular or infragranular neurones. In addition, in the visual cortex, zinc-rich systems appear to converge on Oc2L. Zinc-rich connections appear as an extensive, highly organised association system. PMID- 11906786 TI - Electrophysiological and morphological properties of cell types in the chick neostriatum caudolaterale. AB - The neostriatum caudolaterale, in the chick also referred to as dorsocaudal neostriatal complex, is a polymodal associative area in the forebrain of birds that is involved in sensorimotor integration and memory processes. We have used whole-cell patch-clamp recordings in chick brain slices to characterize the principal cell types of the neostriatum caudolaterale. Electrophysiological properties distinguished four classes of neurons. The morphological characteristics of these classes were examined by intracellular injection of Lucifer Yellow. Type I neurons characteristically fired a brief burst of action potentials. Morphologically, type I neurons had large somata and thick dendrites with many spines. Type II neurons were characterized by a repetitive firing pattern with conspicuous frequency adaptation. Type II neurons also had large somata and thick dendrites with many spines. There was no clear morphological distinction between type I and type II neurons. Type III neurons showed high frequency firing with little accommodation and a prominent time-dependent inward rectification. They had thin, sparsely spiny dendrites and extensive local axonal arborizations. Electrophysiological and morphological properties indicated them as being interneurons. Type IV neurons had a longer action potential duration, a larger input resistance, and a longer membrane time constant than the other classes. Type IV neurons had small somata and short dendrites with few spines. The long axon collaterals of neurons in all spiny cell classes (types I, II, IV) followed similar patterns, suggesting that neurons from all these types can contribute to the projections of the neostriatum caudolaterale to sensory, limbic and motor areas. The electrophysiological and anatomical characterization of the major classes of neurons in the caudal forebrain of the chick provides a framework for the investigation of sensorimotor integration and learning at the cellular level in birds. PMID- 11906787 TI - Effects of early environment on field CA3a pyramidal neuron morphology in the guinea-pig. AB - There is evidence that early environmental conditions have profound effects on the morphology of the dentate granule cells. The aim of the present study was to obtain information about the effects of early environment on neuron morphology in the hippocampal field CA3, a structure closely linked to the dentate gyrus. The dendritic trees and the somata of field CA3a pyramidal neurons were quantified in Golgi-stained brains of guinea-pigs of both sexes raised in either a social or an isolated environment. Two pyramidal neuron types were found in CA3a, characterized by either a long or a short shaft. Environment affected the apical tree of the long-shaft neurons only in males and that of the short-shaft neurons in both sexes. In isolated males the long-shaft neurons had a decrease in the number of dendritic intersections (62-82%), branching points (76%) and length (71%) in the middle third of the apical tree. The short-shaft neurons had a decrease in the number of intersections at two distal levels only in both isolated males (26, 83%) and females (77, 82%). The shaft spine density was affected by environment in the long-shaft neurons of males only, with a density increase (110%) in isolated males. In both sexes the basal tree of only the long shaft neurons was affected by environment. Isolated males had a decrease in the number of dendritic intersections (65-88%), primary dendrites (80%) and dendritic length (88%) and isolated females had a decrease in the number of intersections (51-89%), branching points (77%) and dendritic length (85%). The soma major axis of only the long-shaft neurons was affected by environment with a reduction in isolated males (90%) but an increase in isolated females (111%). These results demonstrate dendritic atrophy of CA3a pyramidal neurons following early isolation and a different reactivity to environment of the two CA3a pyramidal neuron types, their apical and basal trees and the two sexes. The dendritic atrophy of CA3a neurons caused by isolation is likely to be associated with an impairment in the physiology of the hippocampal formation and in the forms of memory in which the hippocampal formation plays a major role. PMID- 11906788 TI - 17beta-estradiol enhances cortical cholinergic innervation and preserves synaptic density following excitotoxic lesions to the rat nucleus basalis magnocellularis. AB - Estradiol exerts beneficial effects on neurodegenerative disorders associated with the decline of cognitive performance. The present study was designed to further investigate the effect of 17beta-estradiol on learning and memory, and to evaluate its neuroprotective action on cholinergic cells of the nucleus basalis magnocellularis, a neural substrate of cognitive performance. Female rats were ovariectomized at an age of 6 months. Three weeks later they received injections of either a mid-physiological dose of 17beta-estradiol or vehicle (oil), every other day for 2 weeks. The effect of estradiol on cognitive performance was tested in two associative learning paradigms. In the two-way active shock avoidance task estradiol-replaced animals learned significantly faster, while in the passive shock avoidance test no differences were observed between the experimental groups. Subsequent unilateral infusion of N-methyl-D-aspartate in the nucleus basalis magnocellularis resulted in a significant loss of cholinergic neurons concomitant with the loss of their fibers invading the somatosensory cortex. Estradiol treatment did not affect the total number of choline acetyltransferase-immunoreactive neurons and their coexpression of the p75 low affinity neurotrophin receptor either contralateral or ipsilateral to the lesion. In contrast, cholinergic fiber densities in estradiol-treated animals were greater both in the contralateral and ipsilateral somatosensory cortices as was detected by quantitative choline-acetyltransferase and vesicular acetylcholine transporter immunocytochemistry. However, estradiol treatment did not affect the lesion-induced relative percentage loss of cholinergic fibers. A significant decline of synaptophysin immunoreactivity paralleled the cholinergic damage in the somatosensory cortex of oil-treated animals, whereas an almost complete preservation of synaptic density was determined in estradiol-treated rats. Our results indicate that estradiol treatment enhances the cortical cholinergic innervation but has no rescuing effect on cholinergic nerve cells in the basal forebrain against excitotoxic damage. Nevertheless, estradiol may restore or maintain synaptic density in the cerebral cortex following cholinergic fiber loss. This estradiol effect may outweigh the lack of cellular protection on cholinergic cells at the functional level. PMID- 11906789 TI - A novel choline-sensitive nicotinic receptor subtype that mediates enhanced GABA release in the chick ventral lateral geniculate nucleus. AB - Nicotinic acetylcholine receptors modulate the release of GABA, glutamate, acetylcholine and dopamine in the brain. Here we describe a novel choline sensitive nicotinic acetylcholine receptor that mediates enhanced GABA release in the chick ventral lateral geniculate nucleus. Whole-cell recordings in slices demonstrated that choline (0.03-10 mM), generally considered an alpha7-selective agonist, and carbachol (3-300 microM), a non-selective cholinergic agonist, both increased the frequency of spontaneous GABAergic events in ventral lateral geniculate nucleus neurons. Tetrodotoxin (0.5 microM) partially reduced responses to carbachol, but eliminated responses to choline. During long-term (5 min) exposure to choline the GABA enhancement was maintained until choline was washed out. Choline (300 microM) enhanced the frequency of spontaneous GABAergic events by 4.28-fold in control artificial cerebrospinal fluid. This choline-mediated enhancement was significantly reduced by the following nicotinic acetylcholine receptor antagonists: 1 microM dihydro-beta-erythroidine (1.49-fold increase, P<0.001), 1 microM methyllycaconitine (1.53-fold, P<0.001) and 0.2 microM alpha conotoxin ImI (1.84-fold, P<0.001). In contrast, no significant change was seen in the presence of 0.1 microM dihydro-beta-erythroidine, 0.1 microM methyllycaconitine, 0.1 microM alpha-bungarotoxin, 0.1 microM alpha-conotoxin MII, 0.1 microM kappa-bungarotoxin, or 1 microM alpha-conotoxin AuIB. These results indicate that choline, at concentrations as low as 100 microM, activates a nicotinic acetylcholine receptor that is distinct from the classical alpha7 nicotinic acetylcholine receptors previously known to be activated by choline. PMID- 11906791 TI - Parcellation of the human pretectal complex: a chemoarchitectonic reappraisal. AB - The pretectum is composed of numerous small nuclei that control various oculomotor functions. In all the non-human mammals investigated, the different pretectal nuclei have been named uniformly according to their structural and functional homology. However, the human pretectal nuclei still bear their traditional, in most cases misleading, nomenclature.In order to reveal the presumed chemoarchitectonic similarities between human and non-human pretectal nuclei, neuropeptide Y (NPY)- and vasoactive intestinal polypeptide (VIP) immunohistochemistry was performed in the human pretectum, after being utilised successfully for the identification of different pretectal nuclei in the cat. No VIP neurones were observed in the human pretectal area, but numerous NPY cells were found in the 'nucleus lentiformis', and in the anterior bulge of the pretectum, while the 'nucleus sublentiformis' contained an abundant NPY fibre network. Some NPY neurones were present in the 'principal pretectal nucleus' as well. The olivary pretectal nucleus possessed NPY fibres, too. In the accessory optic system, the lateral terminal nucleus contained both NPY and VIP neurones, while in the dorsal terminal nucleus only NPY neurones were found. Our chemoanatomical findings were compared to the standard cytoarchitecture as well. Based on the homotopies in the spatial distribution pattern of NPY neurones in the cat and human pretectum, the current, widely accepted non-human anatomical nomenclature was applied to the morphologically homologous nuclei of the human pretectum. Accordingly, the 'nucleus lentiformis' (which contains numerous NPY cells) corresponds to the nucleus of the optic tract, the 'nucleus sublentiformis' (containing a dense network of NPY fibres) to the posterior pretectal nucleus, and the 'nucleus of the pretectal area' corresponds to the medial pretectal nucleus. We identified the anterior part of the pretectum as the human equivalent of the anterior pretectal nucleus in non-humans, including its two compact and reticular subdivisions. In addition, two accessory optic nuclei were verified chemoarchitectonically in the human brain. PMID- 11906790 TI - The neurochemical characterisation of hypothalamic pathways projecting polysynaptically to brown adipose tissue in the rat. AB - The identification of leptin and a range of novel anorectic and orexigenic peptides has focussed attention on the neural circuitry involved in the genesis of food intake and the reflex control of thermogenesis. Here, the neurotropic virus pseudorabies has been utilised in conjunction with the immunocytochemical localisation of a variety of neuroactive peptides and receptors to better define the pathways in the rat hypothalamus directed polysynaptically to the major thermogenic endpoint, brown adipose tissue. Infected neurones were detected initially in the stellate ganglion, then in the spinal cord followed by the appearance of third-order premotor neurones in the brainstem and hypothalamus. Within the hypothalamus these were present in the paraventricular nucleus, lateral hypothalamus, perifornical region, and retrochiasmatic nucleus. At slightly longer survival times virus-infected neurones appeared in the arcuate nucleus and dorsomedial hypothalamus. Neurones in the retrochiasmatic nucleus and in the adjacent lateral arcuate nucleus which project to the brown adipose tissue express cocaine- and amphetamine-regulated transcript, pro-opiomelanocortin and leptin receptors. Neurones in the lateral hypothalamus, a site traditionally associated with the promotion of feeding, project to brown adipose tissue and large numbers of these contained melanin-concentrating hormone and orexin A and B. These data provide part of an anatomical framework which subserves the regulation of energy expenditure. PMID- 11906792 TI - Chemical anoxia activates ATP-sensitive and blocks Ca(2+)-dependent K(+) channels in rat dorsal vagal neurons in situ. AB - The contribution of subclasses of K(+) channels to the response of mammalian neurons to anoxia is not yet clear. We investigated the role of ATP-sensitive (K(ATP)) and Ca(2+)-activated K(+) currents (small conductance, SK, big conductance, BK) in mediating the effects of chemical anoxia by cyanide, as determined by electrophysiological analysis and fluorometric Ca(2+) measurements in dorsal vagal neurons of rat brainstem slices. The cyanide-evoked persistent outward current was abolished by the K(ATP) channel blocker tolbutamide, but not changed by the SK and BK channel blockers apamin or tetraethylammonium. The K(+) channel blockers also revealed that ongoing activation of K(ATP) and SK channels counteracts a tonic, spike-related rise in intracellular Ca(2+) ([Ca(2+)](i)) under normoxic conditions, but did not modify the rise of [Ca(2+)](i) associated with the cyanide-induced outward current. Cyanide depressed the SK channel mediated afterhyperpolarizing current without changing the depolarization-induced [Ca(2+)](i) transient, but did not affect spike duration that is determined by BK channels. The afterhyperpolarizing current and the concomitant [Ca(2+)](i) rise were abolished by Ca(2+)-free superfusate that changed neither the cyanide induced outward current nor the associated [Ca(2+)](i) increase. Intracellular BAPTA for Ca(2+) chelation blocked the afterhyperpolarizing current and the accompanying [Ca(2+)](i) increase, but had no effect on the cyanide-induced outward current although the associated [Ca(2+)](i) increase was noticeably attenuated. Reproducing the cyanide-evoked [Ca(2+)](i) transient with the Ca(2+) pump blocker cyclopiazonic acid did not evoke an outward current. Our results show that anoxia mediates a persistent hyperpolarization due to activation of K(ATP) channels, blocks SK channels and has no effect on BK channels, and that the anoxic rise of [Ca(2+)](i) does not interfere with the activity of these K(+) channels. PMID- 11906793 TI - Neurturin enhances the survival of axotomized retinal ganglion cells in vivo: combined effects with glial cell line-derived neurotrophic factor and brain derived neurotrophic factor. AB - In the present study we localized glial cell line-derived neurotrophic factor (GDNF), and the high affinity receptor for GDNF (GFRalpha-1) in the rat retina. We also examined the effects of neurturin on the survival of axotomized retinal ganglion cells (RGCs) and compared neurturin-mediated RGC rescue to GDNF and brain-derived neurotrophic factor (BDNF) neuroprotection. We administered combined injections of neurturin with BDNF or GDNF in order to determine if these factors rescue RGCs by different mechanisms. GDNF immunoreactivity was localized to RGCs, photoreceptors, and retinal pigment epithelial cells. GFRalpha-1 immunoreactivity was localized to RGCs, Muller cells, and photoreceptors. RGC densities in control retinas decreased from the original value of 2481+/-121 (RGCs/mm(2)+/-S.D.) to 347+/-100 at 14 days post-axotomy. Neurturin treatment significantly increased RGC survival after axotomy (745+/-94) similar to GDNF (868+/-110). BDNF treatment resulted in higher RGC survival (1109+/-156) than either neurturin or GDNF. Combined administration of neurturin with BDNF had additive effects on the survival of axotomized RGCs (1962+/-282), similar to combined administration of GDNF and BDNF (1825+/-269). Combined administration of neurturin and GDNF (1265+/-178) had an enhanced effect on RGC survival. These results suggest that neurturin, GDNF, and BDNF act independently to rescue injured RGCs. Our results also suggest that RGCs and retinal Muller cells may be responsive to GDNF because they both express GFRalpha-1. The present findings have implications for the rescue of injured retinal ganglion cells, as well as other CNS neurons that are responsive to neurturin, GDNF, and BDNF, including midbrain dopaminergic neurons and motor neurons. PMID- 11906794 TI - The expression of Fos-labeled spinal neurons in response to colorectal distension is enhanced after chronic spinal cord transection in the rat. AB - The present study used Fos-like immunoreactivity to examine neuronal activation in response to colorectal distension in rats at 1 day or 30 days following spinal cord transection or sham transection. Fifty-five Wistar rats were anesthetized and an incision was made to expose the T(5) spinal segment. The dura was reflected away in all rats and a complete transection at the rostral end of the T(5) segment was given to the lesioned group. At 1 day (acute) or 30 days (chronic) post-surgery, conscious rats were subjected to a 2 h period of intermittent colorectal distension. Rats were perfused and spinal segments L(5) S(2) were removed and processed for Fos-like immunoreactivity. Spinal cord transection alone had no effect on Fos-labeling in either acute or chronic rats. In acute rats, colorectal distension produced significant increases in Fos labeling in the superficial and deep dorsal horn regions. In chronic rats, colorectal distension produced a three-fold increase in Fos-labeled neurons that was manifest throughout all laminar regions. These results indicate that the number of neurons expressing Fos in response to colorectal distension is much greater after a chronic spinal cord transection than after an acute transection. Since Fos is an indicator of neuronal activation, the results show that many more neurons become active in response to colorectal distension following a chronic spinal injury. This suggests that a functional reorganization of spinal circuits occurs following chronic spinal cord transection. This may ultimately result in altered visceral and somatic functions associated with spinal cord injury in humans. PMID- 11906797 TI - Injection risk behaviors at the first and at the most recent injections among drug users. AB - Risk behaviors at the first intravenous substance injection are unknown. A structured questionnaire was used to investigate the circumstances of the first injection and the changes in risk behaviors between the first and the most recent injections in a group of 143 intravenous drug users (IDUs). When they first injected most subjects were not alone, the initiator was an IDU (94%) who prepared the injection (76%) and did it (79%). The proportions of IDUs sharing preparation equipment (58 vs. 14%), borrowing (23 vs. 2%) and lending injecting equipment (26 vs. 4%) decreased between the first and the most recent injection. PMID- 11906795 TI - Differential screening-selected gene aberrative in neuroblastoma protein modulates inflammatory pain in the spinal dorsal horn. AB - Differential screening-selected gene aberrative in neuroblastoma (DAN) belongs to a novel gene family that includes the Xenopus head-inducing factor, Cerberus and the dorsalizing factor, Gremlin. It has been suggested that members of this family control diverse processes in growth, development and the cell cycle.Here, we demonstrate that the DAN protein is produced in the small neurons of the dorsal root ganglion and is transported to the nerve terminals in the spinal dorsal horn in adult rats. Furthermore, intrathecal injection of an antibody to the DAN protein suppressed inflammatory pain caused by the introduction of complete Freund's adjuvant or carrageenan into the rat hindpaw. The amount of mRNA for DAN in dorsal root ganglion neurons and of its expressed protein in the spinal dorsal horn were both increased in inflammatory models.Together, these data suggest that the DAN protein may be a novel neuromodulator in primary nociceptive nerve fibers. PMID- 11906798 TI - A controlled trial of buprenorphine treatment for opium dependence: the first experience from Iran. AB - AIMS: To evaluate the effect of a 4 mg/day sublingual dose of buprenorphine in the maintenance treatment of opium dependence in comparison with a 1 mg/day dose over an 18-week treatment period. As a secondary objective, the results were determined concurrently for subjects treated with a 2 mg/day dose. DESIGN: Subjects were assigned randomly to three dosage groups. PARTICIPANTS: 330 consecutive (320 men and 10 women) opium addicts who met the DSM-IV criteria for opioid dependence and were seeking treatment. INTERVENTION: Subjects received a 1, 2 or 4 mg/day dose of buprenorphine and were treated in an outpatient clinic where they also received a weekly 1-hour clinical counseling session. MEASUREMENTS: Addiction Severity Index, retention in treatment, and illegal opioid use as determined by random urine testing. FINDINGS: The mean age was 37.5 years (SD=11.4, range 19-72). Overall, 194 (58.8%) of the patients completed the 18 week study. Completion rates by dosage groups were 47.3% for the 1 mg group, 58.2% for the 2 mg group and 70.9% for the 4 mg group (chi(2)=12.7, df=2, P=0.0017). CONCLUSIONS: The results support the efficacy and safety of buprenorphine for opium addiction and suggest that an adequate dose of buprenorphine would help to increase the success rate. PMID- 11906799 TI - MDMA ('ecstasy') use, and its association with high risk behaviors, mental health, and other factors among gay/bisexual men in New York City. AB - This study assesses patterns of use of methylenedioxymethamphetamine (MDMA or 'ecstasy'), and the characteristics of users, in a sample of 733 men who have sex with men (MSM) in New York City. Among respondents, 13.7% reported using MDMA in the past 6 months, with mean frequency of use of 6.24 times in that period. MDMA users were found to be younger, less educated, to have had more male partners, more one night stands with men, more visits to bars or clubs and sex clubs or bathhouses, to have unprotected anal sex with a male, to be likely to have been the victim of physical domestic violence, to have more gay/bisexual friends, to have disclosed their sexual orientation to more friends, family members, and coworkers, and to have higher levels of gay community participation and affiliation. Among MDMA users, higher frequency of MDMA use was associated with being younger, having more visits to bars or clubs, more gay/bisexual friends, and having an HIV negative test result or never having been tested. MDMA users thus constitute a group at risk for sexually transmitted diseases, including HIV, and other problems. The data suggest that MDMA use is associated with being more 'out', which may be advantageous in helping gay men deal with harmful psychological effects of stigma, but may place individuals in settings that expose them to MDMA. These men have also presumably already been well exposed to safer sex messages within the gay community, thus raising challenges for interventions aimed at prevention, as well as opportunities (e.g. MSM and community specific interventions) that need to be further explored. PMID- 11906800 TI - Exposure opportunity as a mechanism linking youth marijuana use to hallucinogen use. AB - The aim of this study is to shed light upon an observed association between the use of marijuana and hallucinogens (e.g. LSD), with a specific focus on the idea that two separate mechanisms might link marijuana use to hallucinogen use: (1) greater hallucinogen exposure opportunity for marijuana users versus nonusers; (2) increased probability of hallucinogen use for marijuana users versus nonusers, once the opportunity to use hallucinogens has occurred. This work is based on a novel analysis of retrospective, self-report data from more than 40000 young participants in the 1991-1994 National Household Surveys on Drug Abuse (NHSDA), with discrete time survival analysis models. Youths who had used marijuana were substantially more likely than nonusers to have the opportunity to use hallucinogens (estimated unadjusted relative risk, uRR=16.3; 95%. Confidence interval (CI)=14.3-18.6). Once an opportunity to use hallucinogens occurred, marijuana users were more likely than nonusers to initiate hallucinogen use (uRR=12.6; 95% CI=9.0-17.6). This evidence provides a more complete view of interconnections between marijuana use and hallucinogen use, and helps to clarify the pivotal role for drug exposure opportunities. Important next steps will be to understand what accounts for variation in the exposure opportunities experienced by marijuana users, and to understand why some marijuana users do not progress even when they have a chance to do so. PMID- 11906801 TI - Differential effects of alcohol, cocaine, and opioid abuse on event-related potentials recorded during a response competition task. AB - The present study examined the abilities of cocaine-dependent and opioid dependent patients and healthy, non-dependent volunteers to execute a task requiring rapid shifts in cognitive set. Sixty-six residential treatment program patients, characterized by a history of either cocaine (n=37) or opioid (n=29) dependence, and 18 non-drug-dependent community volunteers were evaluated. The task involved the execution of right- or left-hand button press responses that were either spatially-compatible or incompatible with a directional cue. Performance and event-related electroencephalographic (EEG) activity were recorded throughout the task. Analyses revealed that button press responses were significantly slower and less accurate for all of the groups when spatial conflict was introduced. The amplitude of a slow EEG potential (SP), emerging approximately 500 ms after stimulus onset, showed the normal effect of spatial conflict for the opioid-dependent and non-dependent groups, but not for subjects in the cocaine-dependent group. Correlational analyses restricted to data from the cocaine-dependent group showed that this abnormal SP amplitude was not related to the quantity, frequency, or recency of their cocaine use; it only correlated with their comorbid alcohol use. The results of the major analyses, in combination with the results of a neuroanatomical localization algorithm applied to the SP data, suggest that comorbid alcohol use disrupts normal task-related activation of the anterior cingulate, cerebellum, and prefrontal cortex. The present results have implications for the assessment of specific cognitive problems which could foster drug abuse. PMID- 11906802 TI - Use, abuse and dependence of ecstasy and related drugs in adolescents and young adults-a transient phenomenon? Results from a longitudinal community study. AB - OBJECTIVE: To determine incidence and patterns of natural course of ecstasy/stimulant/hallucinogen (ESH) use and disorders as well as cohort effects in a community sample of adolescents and young adults. METHOD: Cumulative incidence and patterns of ecstasy use and disorders were examined in a prospective longitudinal design (mean follow-up period=42 months) in a representative sample (N=2446) aged 14-24 years at the outset of the study. Patterns of DSM-IV defined ESH use, abuse and dependence were assessed with the Munich Composite International Diagnostic Interview (M-CIDI). RESULTS: (1) Cumulative lifetime incidence for use of ESH at second follow-up: 9.1%, 1.0% for abuse, 0.6% for dependence; (2) men used and abused ESH more often than women; (3) the younger birth cohort (1977-81) tended to start earlier with substance (ab)use compared to the older birth cohort (1970-77); (4) use of ESH was associated with increasing rates of concomitant use of other licit and illicit drugs; (5) the majority of the lifetime ESH users without disorder had stopped to use these substances and not consumed them during the 12 months preceding the second follow-up; (6) those who had stopped to take ecstasy and related drugs at follow-up also took other illicit drugs less often than those who continued to consume ESH. CONCLUSIONS: Use of designer drugs is widespread in our sample, but the probability of developing use disorders is fairly low (1.6%). The majority of the ESH users stopped their use spontaneously in their twenties (80% of the prior users without disorder, 67% of the prior abusers), but 50% of those that once had fulfilled DSM-IV criteria of dependence continued to use these substances. PMID- 11906804 TI - Ketoconazole increases cocaine and opioid use in methadone maintained patients. AB - Stress plays an important role in substance abuse problems. For example, in studies with rodents stress induces reinstatement of opioid and cocaine self administration. In addition, attenuation of the stress response by pharmacological adrenalectomy using ketoconazole, a cortisol synthesis inhibitor, reduces cocaine self-administration in rodents. In contrast, studies in primates and humans have produced conflicting results using cortisol synthesis inhibitors for attenuating cocaine-related behaviors and subjective effects. To explore the treatment implications of these findings, ketoconazole's (600-900 mg daily) ability to reduce heroin and cocaine use was compared with placebo in 39 methadone maintained patients with a history of cocaine abuse or dependence during a 12-week double blind trial. Contrary to the predicted effects, both heroin and cocaine use increased after patients were stabilized on methadone and ketoconazole. Depressive and withdrawal symptoms improved no more with ketoconazole than with placebo treatment, and side effects were greater on ketoconazole than placebo. As reported before with methadone treatment, morning cortisol levels were significantly lower than normal values throughout the clinical trial, but were not lower with ketoconazole than placebo treatment. Thus, in agreement with the negative results from acute dosing studies in primates and humans, chronic ketoconazole treatment does not appear to reduce cocaine or opioid use in humans maintained on methadone. PMID- 11906803 TI - Neuropsychological performance of individuals dependent on crack-cocaine, or crack-cocaine and alcohol, at 6 weeks and 6 months of abstinence. AB - BACKGROUND: Little data exist on the neuropsychological effects of crack-cocaine dependence or crack-cocaine and alcohol dependence. This study examined cognitive function in abstinent crack dependent and crack and alcohol dependent individuals at 6 weeks and 6 months abstinence. METHODS: a comprehensive neuropsychological battery, including the MicroCog computerized assessment, was administered to 20 abstinent crack dependent subjects, 37 abstinent crack and alcohol dependent subjects, and 29 normal controls. Depression was examined as a covariate, and the association between substance use variables and neuropsychological performance was examined. RESULTS: the two substance dependent groups had similar neuropsychological profiles at 6 weeks abstinent, with both groups exhibiting significant cognitive impairment in a wide range of functions compared to controls. The substance dependent groups were still impaired significantly at 6 months of abstinence. Only mild effects of depression on neuropsychological performance were observed. CONCLUSIONS: crack dependence and crack and alcohol dependence may lead to severe and persistent neuropsychological deficits over a wide range of domains. The strongest predictor of brain damage associated with substance dependence in this sample was dose (particularly quantity and duration of peak dose). PMID- 11906805 TI - Evaluation of an alternative program for MMTP drop-outs: impact on treatment re entry. AB - INTRODUCTION: Retention in a Methadone Maintenance Treatment Program (MMTP) is predictive of abstaining from heroin and has other benefits. Many individuals leave treatment before they experience these positive outcomes. OBJECTIVE: This research project targeted MMTP drop-outs with an intervention designed to assist them in returning to drug treatment. METHODS: Subjects who had left MMTP within the prior 12 months were randomly assigned to intervention or comparison groups. The 3-month long intervention consisted of street outreach, cognitive behavioral groups, and individual counseling. Data were analyzed for 175 subjects who were out of treatment at baseline and who returned for a 6-month follow-up interview (Intervention group, N=111; Comparison group, N=64). RESULTS: A total of 87% of subjects assigned to the intervention condition participated in at least one component. Intervention subjects who attended two or more cognitive behavioral group sessions were more likely than those who attended 0-1 sessions or those in the comparison group to have returned to treatment during the 6 month follow up time period (72 vs. 53 vs. 50%, respectively, P<0.05, chi square test). CONCLUSION: MMTP drop-outs need not be lost to the drug treatment system if special efforts are made to engage them in interventions developed to encourage treatment re-entry. PMID- 11906806 TI - High-risk behaviors associated with transition from illicit non-injection to injection drug use among adolescent and young adult drug users: a case-control study. AB - OBJECTIVE: The goal of our study was to elucidate characteristics of persons likely to transition into injection drug use so that an identifiable group with high-risk for blood-borne infection may be targeted for interventions. METHODS: An age-matched case-control analysis was performed from a cohort study in Baltimore, 1997-1999, of street-recruited non-injection and injection drug users (IDUs), aged 15-30. Cases were IDUs injecting < or = 2 years and controls were age-matched persons who used non-injection heroin, cocaine or crack. At baseline, all were interviewed about prior year-by-year behaviors; analysis using conditional logistic regression was based on information for the year prior to injection onset for the case and the same calendar time for the controls as well as recent behaviors for both groups. RESULTS: Of 270 participants, most were African American (78%), female (61%), and HIV seroprevalence was 7% at baseline. IDUs were significantly more likely than controls to be non-African American (adjusted odds ratio (AOR)=0.09) and report high school dropout (AOR=2.32), early sex-trading (AOR=2.72), and recent violence victimization (AOR=9.28). CONCLUSION: Given that new injectors are at high-risk for HIV and hepatitis yet difficult to reach for prevention efforts, our data suggest some categories to use to target non-injectors who are likely to transition into injection use. PMID- 11906807 TI - Predictors of internalizing and externalizing problems among children of cocaine and opiate dependent parents. AB - We tested associations in structural models among parent individual problems (severity of drug problems, medical problems, psychiatric symptoms), family problems, and children's internalizing and externalizing problems. Results were compared for cocaine versus opiate dependent parents, mothers versus fathers, boys versus girls, and older versus younger children. Cocaine and opiate dependent parents in treatment (N=211) were interviewed about their substance use, psychiatric symptoms, and interpersonal problems and completed a measure of family problems. Parents also rated children's internalizing and externalizing problems. In structural models controlling for the significant correlations between parent and family problems and between children's internalizing and externalizing problems, family problems but not individual parent problems predicted children's internalizing and externalizing symptoms. Models were similar across all groups compared with the exception of parent gender, with significant relations between parent and family problems for mothers but not for fathers. In addition, older girls were more deviant relative to their same-age and gender peers than the younger girls and boys. These results suggest that the personal problems of drug dependent mothers may influence children's problems indirectly by increasing family problems. For drug dependent fathers, family problems were an independent predictor of children's problems. PMID- 11906808 TI - MMPs in the eye: emerging roles for matrix metalloproteinases in ocular physiology. AB - Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes that function to maintain and remodel tissue architecture. Their substrates represent an astounding variety of extracellular matrix components, secreted cytokines and cell surface molecules, and they have been implicated in a wide range of processes and diseases. To date MMPs have been found in virtually every tissue of the eye under conditions of health and disease. Although their functions in vivo remain poorly understood, it is clear they impact on essentially every aspect of eye physiology. This chapter reviews the expanding literature on MMPs in the eye and attempts to place it in the context of basic MMP biology. A general overview of MMP functions is presented first, and then the discussion moves to examples of possible MMP roles in two eye structures. For the cornea, we present recent work on the roles of MMPs during various aspects of wound healing. For the retina, we describe the activities of MMPs in specific disease states from which common principles may emerge. PMID- 11906809 TI - Microemulsions as ocular drug delivery systems: recent developments and future challenges. AB - Eye drops are the most used dosage form by ocular route, in spite of low bioavailability and the pulsed release of the drug. However, due to their intrinsic properties and specific structures, the microemulsions are a promising dosage form for the natural defence of the eye. Indeed, because they are prepared by inexpensive processes through autoemulsification or supply of energy, and can be easily sterilized, they are stable and have a high capacity of dissolving the drugs. The in vivo results and preliminary studies on healthy volunteers have shown a delayed effect and an increase in the bioavailability of the drug. The proposed mechanism is based on the adsorption of the nanodroplets representing the internal phase of the microemulsion, which constitutes a reservoir of the drug on the cornea and should then limit their drainage. PMID- 11906810 TI - Disc haemorrhages, precursors of open angle glaucoma. AB - In a long-term study of 1270 patients with at least one of the findings, open angle glaucoma, disc haemorrhages or retinal vein occlusions, disc haemorrhages were witnessed in approximately 20% of the cases with open-angle glaucoma, and were a precursor of glaucomatous disc changes and associated visual field defects. Disc haemorrhages also preceded a rising intraocular pressure (IOP) in the destructive process among open-angle glaucoma cases. Similar glaucomatous development appears among cases independent of IOP or detection of exfoliation syndrome. Retinal vein occlusions and disc haemorrhages behave similarly with respect to glaucoma. The large number of transgressions is an argument against dividing glaucoma into different types. These findings support a vascular genesis to open-angle glaucoma, presented in earlier epidemiological studies. PMID- 11906811 TI - Retinal transplantation--advantages of intact fetal sheets. AB - Retinal transplantation aims to prevent blindness and to restore eyesight, i.e., to rescue photoreceptors or to replace damaged photoreceptors with the hope of reestablishing neural circuitry. Retinal donor tissue has been transplanted as dissociated cells or intact sheets. A promising experimental paradigm is the subretinal transplantation of sheets of fetal retina with or without its attached retinal pigment epithelium (RPE) into recipient rats with retinal degeneration. As long as healthy RPE either from the host or from the graft is present, such transplants can develop lamination resembling a normal retina. Different methods have been used to demonstrate transplant/host connectivity. In two different rat retinal degeneration models, visually evoked responses can be demonstrated in an area of the superior colliculus corresponding to the placement of the transplant in the retina. In summary, sheets of fetal retina can morphologically repair an area of a degenerated retina, and there is evidence to suggest that transplants form synaptic connections with the host and restore visual responses in blind rats. PMID- 11906812 TI - Beyond Mitomycin: TGF-beta and wound healing. AB - The introduction of the anti-cancer drugs Mitomycin and 5-fluorouracil as anti scarring agents within the last decade, has greatly improved surgical results of glaucoma filtration surgery. However, a number of problems associated with their use have emerged. At the same time, the transforming growth factor-beta (TGF beta) has been identified as an important component of wound healing, particularly in the conjunctival scarring response. Recent developments in molecular therapy offer exciting prospects for the modulation of wound healing, specifically those targeting TGF-beta. As TGF-beta is such a potent stimulant of scarring, this review examines its biology and role in ocular wound healing and repair, and discusses promising new approaches to modifying its activity. PMID- 11906813 TI - Visual field defects and neural losses from experimental glaucoma. AB - Glaucoma is a relatively common disease in which the death of retinal ganglion cells causes a progressive loss of sight, often leading to blindness. Typically, the degree of a patient's visual dysfunction is assessed by clinical perimetry, involving subjective measurements of light-sense thresholds across the visual field, but the relationship between visual and neural losses is inexact. Therefore, to better understand of the effects of glaucoma on the visual system, a series of investigations involving psychophysics, electrophysiology, anatomy, and histochemistry were conducted on experimental glaucoma in monkeys. The principal results of the studies showed that, (1) the depth of visual defects with standard clinical perimetry are predicted by a loss of probability summation among retinal detection mechanisms, (2) glaucomatous optic atrophy causes a non selective reduction of metabolism of neurons in the afferent visual pathway, and (3) objective electrophysiological methods can be as sensitive as standard clinical perimetry in assessing the neural losses from glaucoma. These experimental findings from glaucoma in monkeys provide fundamental data that should be applicable to improving methods for assessing glaucomatous optic neuropathy in patients. PMID- 11906814 TI - Proliferative vitreoretinopathy: risk factors and pathobiology. AB - Proliferative vitreoretinopathy (PVR) is still a major cause of failure of retinal detachment surgery. Despite a dramatic increase in our pathobiologic knowledge of PVR during the last 10 years, little of this information has been used to modify the surgical management of the disease, and, thus, the anatomic and functional results are still unsatisfactory. Collaborative research involving clinicians and basic researchers must be encouraged. PVR must be considered a multifactorial disease caused by interaction of several cells and intra- and extraocular factors. Therefore, therapeutic options based on the inhibition of one factor or phenomenon may be regarded with scepticism. To prevent PVR, it is necessary to determine the factors involved in its development, and because of its relatively small prevalence, large, prospective, multicenter studies seem necessary. In addition, clinical research must not be underestimated. PVR affects both sides of the retina and the retina itself, a point to which little attention has been paid and that is critical for surgical results. Therefore, a new classification that provides information about clinical relevance, such as the evolutionary stages of the disease (biologic activity) and the degree of surgical difficulty (location of the fibrotic process), seems necessary. PMID- 11906815 TI - Clusterin. AB - Clusterin is an enigmatic glycoprotein with a nearly ubiquitous tissue distribution and an apparent involvement in biological processes ranging from mammary gland involution to neurodegeneration in Alzheimer's disease. Its major form, a 75-80 kDa heterodimer, is secreted and present in physiological fluids, but truncated forms targeted to the nucleus have also been identified. Upregulation of clusterin mRNA and protein levels detected in diverse disease states and in in vitro systems have led to suggestions that it functions in membrane lipid recycling, in apoptotic cell death, and as a stress-induced secreted chaperone protein, amongst others. Recent studies of knockout mice have further complicated the picture by implicating clusterin in exacerbating neuronal death in hypoxia-ischemia. The question of whether clusterin is a multifunctional protein, or deploys a single primary function influenced by cellular context, remains a central issue continuing to stimulate interest in this unusual molecule. PMID- 11906816 TI - RGS2: a multifunctional regulator of G-protein signaling. AB - Regulators of G-protein signaling (RGS) proteins enhance the intrinsic rate at which certain heterotrimeric G-protein alpha-subunits hydrolyze GTP to GDP, thereby limiting the duration that alpha-subunits activate downstream effectors. This activity defines them as GTPase activating proteins (GAPs). As do other RGS proteins RGS2 possesses a 120 amino acid RGS domain, which mediates its GAP activity. In addition, RGS2 shares an N-terminal membrane targeting domain with RGS4 and RGS16. Found in many cell types, RGS2 expression is highly regulated. Functionally, RGS2 blocks Gq alpha-mediated signaling, a finding consistent with its potent Gq alpha GAP activity. Surprisingly, RGS2 inhibits Gs signaling to certain adenylyl cyclases. Like other RGS proteins, RGS2 lacks Gs alpha GAP activity, however it directly inhibits the activity of several adenylyl cyclase isoforms. Targeted mutation of RGS2 in mice impairs anti-viral immunity, increases anxiety levels, and alters synaptic development in hippocampal CA1 neurons. RGS2 has emerged as a multifunctional RGS protein that regulates multiple G-protein linked signaling pathways. PMID- 11906817 TI - Glutamine and its relationship with intracellular redox status, oxidative stress and cell proliferation/death. AB - Glutamine is a multifaceted amino acid used for hepatic urea synthesis, renal ammoniagenesis, gluconeogenesis in both liver and kidney, and as a major respiratory fuel for many cells. Decreased glutamine concentrations are found during catabolic stress and are related to susceptibility to infections. Besides, glutamine is not only an important energy source in mitochondria, but is also a precursor of the brain neurotransmitter glutamate, which is likewise used for biosynthesis of the cellular antioxidant glutathione. Reactive oxygen species, such as superoxide anions and hydrogen peroxide, function as intracellular second messengers activating, among others, apoptosis, whereas glutamine is an apoptosis suppressor. In fact, it could contribute to block apoptosis induced by exogenous agents or by intracellular stimuli. In conclusion, this article shows evidences for the important role of glutamine in the regulation of the cellular redox balance, including brain oxidative metabolism, apoptosis and tumour cell proliferation. PMID- 11906818 TI - ATPase pumps in osteoclasts and osteoblasts. AB - Osteoblasts, osteocytes and osteoclasts are specialised cells of bone that play crucial roles in the formation, maintenance and resorption of bone matrix. Bone formation and resorption critically depend on optimal intracellular calcium and phosphate homeostasis and on the expression and activity of plasma membrane transport systems in all three cell types. Osteotropic agents, mechanical stimulation and intracellular pH are important parameters that determine the fate of bone matrix and influence the activity, expression, regulation and cell surface abundance of plasma membrane transport systems. In this paper the role of ATPase pumps is reviewed in the context of their expression in bone cells, their contribution to ion homeostasis and their relation to other transport systems regulating bone turnover. PMID- 11906819 TI - Prenylcysteine carboxymethyltransferase type III activity is decreased in retinoic acid-treated SH-SY5Y neuroblastoma cells. AB - Prenylcysteine carboxymethyltransferase (pcCMT) is an enzyme that catalyzes the post-translational carboxymethylation of isoprenylated proteins ensuring a more efficient membrane attachment and proper guiding to a specific target membrane. In this paper, we report on modulation of pcCMT activity in retinoic acid (RA) treated SH-SY5Y neuroblastoma cells using N-acetyl-S-farnesyl-L-cysteine (AFC) as artificial methyl acceptor. In addition, the methylation of endogenous proteins was followed by the vapor phase equilibrium assay and the storage phosphor screen (P-screen) technique with S-adenosyl-[3H-methyl] methionine (AdoMet) as methyl donor. Methylation of AFC was reduced to 75% of that of the control, the most prominent decrease being observed with the post-nuclear membrane fraction as enzyme source. With regard to protein methylation both screening methods yielded analogous results showing the [3H]-labeling of endogenous proteins in the 21 25kDa molecular mass (MM) range to be diminished by nearly 50%. This questions the role of protein carboxymethylation as an essential component of the differentiation process in SH-SY5Y neuroblastoma cells. The P-screen technique revealed that the methylation of other molecular mass proteins was also affected. Both S-adenosylhomocysteine (AdoHcy) and AFC (AdoHcy being the most effective) inhibited endogenous methylation. An interesting feature was that AFC inhibited the protein methylation proportionally more effective in RA-treated cells. Finally, the levels of three small guanosine-5'-triphosphate (GTP) binding proteins were screened upon differentiation showing rab3A to be increased while rhoA and H-ras were decreased. PMID- 11906820 TI - The novel human HUEL (C4orf1) protein shares homology with the DNA-binding domain of the XPA DNA repair protein and displays nuclear translocation in a cell cycle dependent manner. AB - We have previously isolated and characterized a novel human gene HUEL (C4orf1) that is ubiquitously expressed in a wide range of human fetal, adult tissues and cancer cell lines. HUEL maps to region 4p12-p13 within the short arm of chromosome 4 whose deletion is frequently associated with bladder and other carcinomas. Here we present the genomic organization, sizes and boundaries of exons and introns of HUEL. The GC-rich upstream genomic region and 5' untranslated region (UTR) together constitute a CpG island, a hallmark of housekeeping genes. The 3250 bp HUEL cDNA incorporates a 1704 bp ORF that translates into a hydrophilic protein of 568-amino acids (aa), detected as a band of approximately 70 kDa by Western blotting. We have isolated the murine homolog of HUEL which exhibits 89% nucleotide and 94% amino acid identity to its human counterpart. The HUEL protein shares significant homology with the minimal DNA binding domain (DNA-BD) of the DNA repair protein encoded by the xeroderma pigmentosum group A (XPA) gene. Other notable features within HUEL include the putative nuclear receptor interaction motif, nuclear localization and export signals, zinc finger, leucine zipper and acidic domains. Mimosine-mediated cell cycle synchronization of PLC/PRF/5 liver cancer cells clearly portrayed translocation of HUEL into the nucleus specifically during the S phase of the cell cycle. Yeast two-hybrid experiments revealed interactions of HUEL with two partner proteins (designated HIPC and HIPB) bearing similarity to a mitotically phosphorylated protein and to reverse transcriptase. Co-immunoprecipitation assays validated the interaction between HUEL and HIPC proteins in mammalian cells. HUEL is likely to be an evolutionarily conserved, housekeeping gene that plays a role intimately linked with cellular replication, DNA synthesis and/or transcriptional regulation. PMID- 11906821 TI - Cloning of a differentially expressed tropomyosin isoform from cultured rabbit aortic smooth muscle cells. AB - The four known tropomyosin genes have highly conserved DNA and amino acid sequences, and at least 18 isoforms are generated by alternative RNA splicing in muscle and non-muscle cells. No rabbit tropomyosin nucleotide sequences are known, although protein sequences for alpha- and beta-tropomyosin expressed by rabbit skeletal muscle have been described. Subtractive hybridisation was used to select for genes differentially expressed in rabbit aortic smooth muscle cells (SMC), during the change in cell phenotype in primary culture that is characterised by a loss of cytoskeletal filaments and contractile proteins. This led to the cloning of a tropomyosin gene predominantly expressed in rabbit SMC during this change. The full-length cDNA clone, designated "rabbit TM-beta", contains an open reading frame of 284 amino acids, 5' untranslated region (UTR) of 117 base pairs and 3' UTR of 79 base pairs. It is closely related to the beta gene isoforms in other species, with the highest homology in DNA and protein sequences to the human fibroblast isoform TM-1 (91.7% identity in 1035 bp and 93.3% identity in the entire 284 amino acid sequence of the protein). It differs from rabbit skeletal muscle beta-tropomyosin (81.7% homology at the protein level) mainly in two regions at amino acids 189-213 and 258-283 suggesting alternative splicing of exons 6a for 6b and 9d for 9a. Since this TM-beta gene was the only gene strongly enough expressed in SMC changing phenotype to be observed by the subtractive hybridisation screen, it likely plays a significant role in this process. PMID- 11906822 TI - Electrocyte (Na(+),K(+))ATPase inhibition induced by zinc is reverted by dithiothreitol. AB - The Mg(2+)-dependent (Na(+),K(+))ATPase maintains several cellular processes and is essential for cell excitability. In view of the importance of the enzyme activity, the interaction and binding affinities to substrates and metal ions have been studied. We determined the effect of Zinc ion (Zn(2+)) on the (Na(+),K(+))ATPase activity present in both conducting (non-innervated) and post synaptic (innervated) membranes of electrocyte from Electrophorus electricus (L.). Zn(2+) is involved in many biological functions and is present in pre synaptic nerve terminals. This metal, which has affinity for thiol groups, acted as a potent competitive inhibitor of (Na(+),K(+))ATPase of both membrane fractions, which were obtained by differential centrifugation of the E. electricus main electric organ homogenate. We tried to recover the enzyme activity using dithiothreitol, a reducing agent. Kinetic analysis showed that dithiothreitol acted as a non-essential non-competitive activator of (Na(+),K(+))ATPase from both membrane fractions and was able to revert the Zn(2+) inhibition at mM concentrations. In the presence of dithiothreitol, this metal behaved as a competitive inhibitor of (Na(+),K(+))ATPase in the non-innervated membrane fractions and presented a non-competitive inhibition of (Na(+),K(+))ATPase in innervated membrane fractions. This difference may be attributed to formation of a Zn-dithiothreitol complex, as well as the involvement of other binding sites for both agents. The consequences of the enzyme inhibition by Zn(2+) may be considered in regard to its neurotoxic effects. PMID- 11906823 TI - Polyunsaturated fatty acid biosynthesis from [1-14C]20:3 n-6 acid in rat cultured Sertoli cells. Linoleic acid effect. AB - Primary culture is a suitable system to study lipid metabolism and polyunsaturated fatty acid biosynthesis. Sertoli cell-enriched preparations were used to determine the fatty acid composition after 5 and 7 days in culture (serum free) as well as the uptake and metabolism of [1-14C]eicosa-8,11,14-trienoic acid. The addition of unlabeled linoleic acid (0.2 and 2.0 microg/ml) was also evaluated. Fatty acid methyl esters derived from cellular lipids were analyzed by gas liquid chromatography and radiochromatography. After 5 days in culture, cells had significantly less 18:2, 20:4, 22:5 and 24:5 and more 18:3, 20:3, 22:4 and 24:4 n-6 fatty acids than non-cultured cells. On day 7, an additional increment in 22:4 n-6 and a decrease in linoleic, gamma-linoleic and 24:4 n-6 fatty acids were observed. The presence of linoleic acid (low dose) produced a significant decrease in saturated and monounsaturated acids and an increase in 18:2, 20:4 and 22:5 n-6 fatty acids. At a high concentration almost all fatty acids belonging to 18:2 n-6 increased significantly. The drop in 20:4 n-6/20:3 n-6 ratio was considered as an indirect evidence of a Delta 5 desaturase activity depression. This assumption was corroborated by studying the transformation of [1-14C]eicosa 8,11,14-trienoic acid into 20:4, 22:4, 22:5, 24:4 and 24:5 n-6 fatty acids. We conclude that Sertoli cells after 7 days in culture evidenced changes in the fatty acid profile similar to those described under fat deprivation. The addition of linoleic acid reverted this pattern and indicated that the Delta 5 desaturase activity is a limiting step in the polyunsaturated fatty acid biosynthesis. PMID- 11906824 TI - Evidence for a dimeric structure of rat liver serine dehydratase. AB - Rat liver serine dehydratase (SDH) is known to be involved in gluconeogenesis. It has long been believed to be a dimeric protein with the subunit molecular weight (M(r)) of 34,000. Recently, sheep liver SDH was reported to be a monomer with a M(r) of 38,000. The native M(r) of rat SDH was only determined by the ultracentrifugation method more than three decades ago, and that of sheep SDH was done by the method of gel chromatography. The primary to quaternary structures of a given enzyme in a specific mammalian organ are usually conserved among various species. The aim of the present investigation is to clarify the structural differences between rat and sheep SDHs. First, we found that the amino acid composition reported for sheep SDH was statistically similar to that of rat SDH. Second, immunoblot analysis using anti-rat SDH IgG as the probe showed the size of sheep SDH to be a M(r) of 30,500, whereas that of SDH was about M(r) of 35,000. On the other hand, the native size of rat SDH was assessed by two methods: (1) the laser light scattering method demonstrated that rat SDH had a M(r) of 66,800, consistent with the previous value (M(r)=64,000); (2) cross linking experiments of the purified rat SDH with dimethyl suberimidate revealed the existence of a dimeric form by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The present results clearly confirm that rat SDH is a dimer, and suggest that sheep SDH is similar to rat SDH immunologically, but with a molecular weight 7500 smaller than reported previously. PMID- 11906825 TI - Melatonin protects against delta-aminolevulinic acid-induced oxidative damage in male Syrian hamster Harderian glands. AB - Effects of the prooxidant delta-aminolevulinic acid (ALA) and the antioxidant melatonin (MEL) were investigated in the male Syrian hamster Harderian gland (HG). Rodent Harderian glands are highly porphyrogenic organs, which may be used as model systems for studying damage by delta-aminolevulinic acid and its metabolites, as occurring in porphyrias. Chronic administration of delta aminolevulinic acid (2 weeks) markedly decreased activities of the porphyrogenic enzymes delta-aminolevulinate synthase (ALA-S) and delta-aminolevulinate dehydratase (ALA-D) and of the antioxidant enzymes superoxide dismutase (SOD), glutathione reductase (GR) and catalase (CAT), whereas porphobilinogen deaminase (PBG-D) remained unaffected. This treatment led to increased lipid peroxidation (LPO) and oxidatively modified protein (protein carbonyl) as well as to morphologically apparent tissue damage. Melatonin also caused decreases in delta aminolevulinate synthase, delta-aminolevulinate dehydratase, superoxide dismutase, glutathione reductase and catalase. Despite lower activities of antioxidant enzymes, lipid peroxidation and protein carbonyl were markedly diminished. The combination of delta-aminolevulinic acid and melatonin led to approximately normal levels of delta-aminolevulinate dehydratase, glutathione reductase, catalase and protein carbonyl, and to rises in superoxide dismutase and porphobilinogen deaminase activities; lipid peroxidation remained even lower than in controls and the appearance of the tissue revealed a protective influence of melatonin. These results suggest that melatonin may have profound effects on the oxidant status of the Harderian gland. PMID- 11906826 TI - Bisphosphonates modulate the effect of macrophage-like cells on osteoblast. AB - Macrophages (MPs) are present in many tissues and have been implicated in the excessive bone resorption seen in patients with skeletal disorders. Our previous studies showed that macrophage-like cells influenced osteoblasts (OB) in co culture, as number and activity of osteoblasts were decreased in co-cultures compared with controls. Macrophages are probable precursors of osteoblasts which have been shown to be inhibited by bisphosphonates (BPs). Bisphosphonates also modulate macrophage and osteoblasts activity. This study investigated whether addition of bisphosphonates to co-cultures of osteoblast and macrophages could reduce or block the adverse effects of macrophages on osteoblasts. The results showed that, compared to controls, fewer osteoblasts were present over time in macrophage/osteoblast co-cultures (at day 12, 15.5 x 10(4) and 8.8 x 10(4); P<0.0001) and that addition of bisphosphonates (10(-9)-10(-5)M) to the co cultures prevented this reduction (P<0.001). Bisphosphonates also elicited an increase in numbers of osteoblast (82%) and restored alkaline phosphatase (ALP) activity, which was reduced by 15% (P approximately equal to 0.05) compared to control levels. The number of macrophages in co-cultures was reduced when bisphosphonates were added (P<0.001) and release of lactate dehydrogenase (LDH) was seen, which was not detectable in control cultures. It therefore, appears that bisphosphonates initiated macrophage death. These results demonstrated that the inhibitory effect of macrophages on osteoblasts in vitro could be overcome by the action of bisphosphonates. These findings have implications for the treatment of skeletal conditions where macrophage-derived cytokines are important, such as arthritis and implant loosening, although it is clearly important to distinguish between those bisphosphonates which enhance synthesis of pro-inflammatory cytokines and those which inhibit such synthesis. PMID- 11906828 TI - Calcium-dependent protein kinases: versatile plant signalling components necessary for pathogen defence. PMID- 11906827 TI - Lead ion effect on creatine kinase: equilibrium and kinetic studies of inactivation and conformational changes. AB - The effects of lead ions on creatine kinase (CK) were studied by measuring activity changes, intrinsic fluorescence spectra and 8-anilo-1 naphthalenesulfonate (ANS)-binding fluorescence along with size-exclusion chromatography (SEC). Below 5 mM Pb(2+) concentration, there was nearly no change of the enzyme activity and a slight change of the ANS-binding fluorescence. The CK activity decreased significantly from 10 to 25 mM Pb(2+) concentrations. No residual activity was observed above 25 mM Pb(2+). The kinetic time courses of inactivity and unfolding were all mono-phase courses with the inactivation rate constants being greater than the unfolding rate constants for the same Pb(2+) concentration. The changes in fluorescence maximum and fluorescence intensity were relatively slow for 40-80 mM Pb(2+) as well as in the initial stage for less than 5 mM Pb(2+), showing that two transition states exist for Pb(2+) induced equilibrium-unfolding curves. The intrinsic fluorescence spectra and ANS-binding fluorescence measurements showed that even for high Pb(2+) concentrations, CK did not fully unfold. Additionally, the SEC results showed that the enzyme molecule still existed in an inactive dimeric state at 20 and 40 mM Pb(2+) solutions. All the results indicated the presence of at least one stable unfolding equilibrium intermediate of CK during Pb(2+) unfolding. PMID- 11906829 TI - Chloroplast biogenesis and function are first in the list of essential Arabidopsis genes. PMID- 11906830 TI - Transcript profiling takes a stroll through the wood. PMID- 11906833 TI - bZIP transcription factors in Arabidopsis. AB - In plants, basic region/leucine zipper motif (bZIP) transcription factors regulate processes including pathogen defence, light and stress signalling, seed maturation and flower development. The Arabidopsis genome sequence contains 75 distinct members of the bZIP family, of which approximately 50 are not described in the literature. Using common domains, the AtbZIP family can be subdivided into ten groups. Here, we review the available data on bZIP functions in the context of subgroup membership and discuss the interacting proteins. This integration is essential for a complete functional characterization of bZIP transcription factors in plants, and to identify functional redundancies among AtbZIP factors. PMID- 11906832 TI - Rings and networks: the amazing complexity of FtsZ in chloroplasts. AB - Bacteria have proteins that can form filaments and rings, and these are thought to be the evolutionary progenitors of actin and tubulin. Plant homologues of the most intensively studied bacterial FtsZ protein are nuclear-encoded by a small gene family, are plastid-bound and participate in the plastid division process. The hypothesis is put forward that FtsZ and other proteins form a filamentous network in plastids, a plastoskeleton, which keeps these organelles in shape and helps them to divide. PMID- 11906834 TI - Galactolipids rule in seed plants. AB - Chloroplast membranes contain high levels of the galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG). The isolation of the genes involved in the biosynthesis of MGDG and DGDG, and the identification of galactolipid-deficient Arabidopsis mutants has greatly facilitated the analysis of galactolipid biosynthesis and function. Galactolipids are found in X-ray structures of photosynthetic complexes, suggesting a direct role in photosynthesis. Furthermore, galactolipids can substitute for phospholipids, as suggested by increases in the galactolipid:phospholipid ratio after phosphate deprivation. The ratio of MGDG to DGDG is also crucial for the physical phase of thylakoid membranes and might be regulated. PMID- 11906835 TI - Engineering crop plants: getting a handle on phosphate. AB - In plant seeds, most of the phosphate is in the form of phytic acid. Phytic acid is largely indigestible by monogastric animals and is the single most important factor hindering the uptake of a range of minerals. Engineering crop plants to produce a heterologous phytase improves phosphate bioavailability and reduces phytic acid excretion. This reduces the phosphate load on agricultural ecosystems and thereby alleviates eutrophication of the aquatic environment. Improved phosphate availability also reduces the need to add inorganic phosphate, a non renewable resource. Iron and zinc uptake might be improved, which is significant for human nutrition in developing countries. PMID- 11906836 TI - Sieve elements caught in the act. AB - Phloem is a puzzling plant tissue owing to the unique natural defence responses of the sieve elements to any kind of mechanical manipulation. Recent non-invasive studies have enabled real-time observation of events in intact sieve tubes, including mass transport, sieve-pore sealing and conformational changes of structural proteins. These studies further highlighted the importance of the symplasmic setting for development and functioning of the sieve elements. Exchange of macromolecules between companion cells and sieve elements is indispensable for the survival of the sieve element, but also seems to be involved in long-distance communication. How the branched plasmodesmata between sieve element and companion cell function as corridors for the passage of macromolecules is an intriguing but unresolved story. PMID- 11906838 TI - International Rice Research Institute: roles and challenges as we enter the genomics era. AB - The International Rice Research Institute (IRRI) was established in 1960 by the Rockefeller (New York, NY, USA) and Ford Foundations (New York, NY, USA) in response to food scarcity problems in the developing world. Today, it is the world's leading international research and training center for rice. Based in the Philippines, with operations in 11 other countries, it is one of 16 Future Harvest Centers funded by the Consultative Group on International Agricultural Research (CGIAR), an association of public and private donor agencies. PMID- 11906837 TI - Nuclear phosphoinositides could bring FYVE alive. AB - Phosphoinositide signalling systems exist in all eukaryotes. A high degree of evolutionary conservation is found at the functional level, but distinct phylogenetic differences are also becoming evident. Although the nuclear phosphoinositide system is likely to be a primordial forerunner of the plasma membrane system, relatively little is known about it. However, nuclear phosphoinositides might have far more diverse roles than hitherto envisaged and interact specifically with regulatory proteins containing phosphoinositide binding domains. A novel family of proteins, so far only identified in plants, display domain structures that might link phosphoinositide metabolism to nuclear function in an unexpected way. PMID- 11906839 TI - Shedding light on proteins, nucleic acids, cells, humans and fish. AB - I was trained as a physicist in graduate school. Hence, when I decided to go into the field of biophysics, it was natural that I concentrated on the effects of light on relatively simple biological systems, such as proteins. The wavelengths absorbed by the amino acid subunits of proteins are in the ultraviolet (UV). The wavelengths that affect the biological activities, the action spectra, also are in the UV, but are not necessarily parallel to the absorption spectra. Understanding these differences led me to investigate the action spectra for affecting nucleic acids, and the effects of UV on viruses and cells. The latter studies led me to the discovery of the important molecular nature of the damages affecting DNA (cyclobutane pyrimidine dimers) and to the discovery of nucleotide excision repair. Individuals with the genetic disease xeroderma pigmentosum (XP) are extraordinarily sensitive to sunlight-induced skin cancer. The finding, by James Cleaver, that their skin cells were defective in DNA repair strongly suggested that DNA damage was a key step in carcinogenesis. Such information was important for estimating the wavelengths in sunlight responsible for human skin cancer and for predicting the effects of ozone depletion on the incidence of non melanoma skin cancer. It took experiments with backcross hybrid fish to call attention to the probable role of the longer UV wavelengths not absorbed by DNA in the induction of melanoma. These reflections trace the biophysicist's path from molecules to melanoma. PMID- 11906840 TI - A systematic review of cytogenetic studies conducted in human populations exposed to cadmium compounds. AB - BACKGROUND: Exposure to cadmium fumes or dusts has been associated with an increased risk of lung cancer and the characterisation of the genotoxic potential of cadmium compounds is, among other possible mechanisms, an important element in the assessment of the carcinogenic hazard of the element. While there is some evidence that in experimental systems, cadmium compounds may exert genotoxic effects, the results of the epidemiological studies having examined cytogenetic endpoints in humans exposed to cadmium appear conflicting. Therefore, a systematic review was undertaken to assess whether a cytogenetic effect of cadmium exposure is supported by the studies with the strongest design. METHODS: The relevant literature was identified through several databases and assessed with a check-list by two reviewers. Causes of heterogeneity between studies were looked for. Results were extracted and the strength of the evidence was evaluated with causality criteria. RESULTS: No studies met the criteria for being considered as very convincing. Several factors were identified that could explain contradictory findings (small sample size, selection bias, insufficient characterisation of exposure, lack of consideration of confounders) but their actual impact could not be conclusively assessed with the published information. Importantly, it should be recognised that the absence of a clear mechanism for the cytogenetic action of cadmium compounds did not allow to select the most appropriate endpoint to be examined. CONCLUSIONS: No clear association between cadmium exposure and cytogenetic endpoint appeared but no definite conclusion can be drawn from the existing studies in humans. Future research efforts should mainly focus on experimental studies to understand how cadmium compounds could produce genotoxic/carcinogenic effects, in order to target the most relevant endpoint to be examined in humans. PMID- 11906841 TI - A role for p53 in the frequency and mechanism of mutation. AB - The tumor suppressor protein, p53, is often referred to as the guardian of the genome. When p53 function is impaired, its ability to preserve genomic integrity is compromised. This may result in an increase in mutation on both a molecular and chromosomal level and contribute to the progression to a malignant phenotype. In order to study the effect of p53 function on the acquisition of mutation, in vitro and in vivo models have been developed in which both the frequency and mechanism of mutation can be analyzed. In human lymphoblastoid cells in which p53 function was impaired, both the spontaneous and induced mutant frequency increased at the autosomal thymidine kinase (TK) locus. The mutant frequency increased to a greater extent in cell lines in which p53 harbored a point mutation than in those lines in which a "null" mutation had been introduced by molecular targeting or by viral degradation indicating a possible "gain-of function" associated with the mutant protein. Further, molecular analysis revealed that the loss of p53 function was associated with a greater tendency towards loss-of-heterozygosity (LOH) within the TK gene that was due to non homologous recombination than that found in wild-type cells. Most data obtained from the in vivo models uses the LacI reporter gene that does not efficiently detect mutation that results in LOH. However, studies that have examined the effect of p53 status on mutation in the adenine phosphoribosyl transferase (APRT) gene in transgenic mice also suggest that loss of p53 function results in an increase in mutation resulting from non-homologous recombination. The results of these studies provide clear and convincing evidence that p53 plays a role in modulating the mutant frequency and the mechanism of mutation. In addition, the types of mutation that occur within the p53 gene are also of importance in determining the mutant frequency and the pathways leading to mutation. PMID- 11906842 TI - International Commission for the Protection of the Environment against Mutagens and Carcinogens: a historical perspective. PMID- 11906845 TI - Harold G. Jacobson of the Bronx. PMID- 11906843 TI - Biomarkers in molecular epidemiology studies for health risk prediction. AB - The field of molecular epidemiology is very promising, as sophisticated techniques are being developed to address etiology, genetic susceptibility and mechanisms for induction of disease. The use of biomarkers plays a key role in these investigations because the information can be used to predict the development of disease and to implement disease prevention programs. However, as emphasized by Frederica P. Perera, the field is strewn with studies either that failed to use validated biomarkers or whose designs did not adequately consider the biology of the endpoints, and the availability of validated biomarkers of health risk is still limited. In this review, we have briefly described the usefulness of certain biomarkers for the documentation of exposure and early biological effects, with special concern for the prediction of cancer. An emphasis is placed on understanding the biological and health significance of biomarkers. By building reliable biomarker databases, a promising future is the integration of information from the genome programs to expand the scientific frontiers on etiology, health risk prediction and prevention of environmental disease. PMID- 11906847 TI - Harold Gordon Jacobson, 1912-2001. PMID- 11906850 TI - Image quality of screening mammography: effect on clinical outcome. PMID- 11906848 TI - Screening mammography: clinical image quality and the risk of interval breast cancer. AB - OBJECTIVE: We evaluated the association between clinical image quality and breast cancer occurrence within 24 months of a negative mammogram. MATERIALS AND METHODS: We identified women with breast cancer who were younger than 40 years old and older and screened from January 1, 1988, through December 31, 1993. We retrospectively assigned Breast Imaging Reporting and Data System (BI-RADS) assessments to their screening mammogram. We classified cancers (invasive or ductal in situ) as "screen-detected" when found after positive assessments (BI RADS codes 3, 4, and 5) and "interval-detected" when found after negative assessments (BI-RADS codes 1 and 2). One reviewer evaluated mediolateral oblique and craniocaudal views for all cancer cases using a 3-point scale (failure, borderline, pass) for each measure of clinical image quality (positioning, breast compression, contrast, exposure, noise, sharpness, artifacts, overall quality). We used separate logistic regression models and evaluated the odds of interval invasive cancer or invasive plus in situ cancer as a function of each measure of quality using "pass" as the referent group. RESULTS: We found 492 screen-detected and 164 interval-detected cancers that met study criteria. Cancer detection (sensitivity) was highest (84%) among patients with proper breast positioning, but when images failed this measure (33.4%), sensitivity fell to 66.3%. After adjustment for age, film date, and breast density, interval-detected invasive cancers were more likely after images failing positioning (odds ratio, 2.57; 95% confidence interval, 1.28-5.52%). Failures in overall quality were also associated with interval cancers when cases of ductal carcinoma in situ (p = 0.037) were included. CONCLUSION: Invasive breast cancer detection by mammography may be improved through attention to correct positioning. PMID- 11906852 TI - Communicating findings of radiologic examinations: whither goest the radiologist's duty? PMID- 11906853 TI - A simple method of capturing PACS and other radiographic images for digital teaching files or other image repositories. AB - OBJECTIVE: We sought to develop an easy-to-use method of capturing and storing radiographic images. CONCLUSION: The method that we developed can capture any digital image-including an image from a picture archiving and communication system (PACS)-using widely available, inexpensive software. Our method is easy to learn, simple to use, and inexpensive to implement. It is adaptable in a wide range of networking environments and can capture and store images rapidly for a variety of uses. It can be used without interfering with clinical workflow at the PACS workstation. PMID- 11906855 TI - Dual-phase helical CT of pancreatic adenocarcinoma: assessment of resectability before surgery. AB - OBJECTIVE: The aim of our study was to prospectively evaluate the accuracy of dual-phase helical CT in the preoperative assessment of resectability in patients with suspected pancreatic cancer using surgical and histopathologic correlation. SUBJECTS AND METHODS: Between January 1999 and December 2000, 76 patients with suspected pancreatic cancer underwent preoperative evaluation and staging with dual-phase helical CT (3-mm collimation for pancreatic phase, 5-mm collimation for portal phase). Iodinated contrast material was injected IV (170 mL at a rate of 4 mL/sec); acquisition began at 40 sec during the pancreatic phase and at 70 sec during the portal phase. Three radiologists prospectively evaluated the imaging findings to determine the presence of pancreatic tumor and signs of unresectability (liver metastasis, vascular encasement, or regional lymph nodes metastasis). The degree of tumor-vessel contiguity was recorded for each patient (no contiguity with tumor, contiguity of < 50%, or contiguity of > or =50%). RESULTS: Thirty-nine patients with pancreatic adenocarcinoma were surgically explored. Curative resections were attempted in 34 patients and were successful in 25. The positive predictive value for resectability was 73.5%. Nine patients considered resectable on the basis of CT findings were found to be unresectable at surgery because of liver metastasis (n = 5), vascular encasement (n = 2), or lymph node metastasis (n = 2). We found that the overall accuracy of helical CT as a tool for determining whether a pancreatic adenocarcinoma was resectable was 77% (30/39 patients). CONCLUSION: Dual-phase helical CT is a useful technique for preoperative staging of pancreatic cancer. The main limitation of CT is that it may not reveal small hepatic metastases. PMID- 11906856 TI - Multidetector CT angiography of pancreatic carcinoma: part I, evaluation of arterial involvement. PMID- 11906858 TI - Multidetector CT angiography of pancreatic carcinoma: part 2, evaluation of venous involvement. PMID- 11906859 TI - Simple pancreatic cysts: CT and endosonographic appearances. AB - OBJECTIVE. This report describes the CT and endoscopic sonographic appearance of simple pancreatic cysts in three adults. CONCLUSION. Simple pancreatic cysts are typically an incidental finding in adults who have no history of pancreatic disease. The imaging characteristics of simple pancreatic cysts on CT and endosonography are similar to those of benign cysts. PMID- 11906860 TI - Gallium uptake in complicated pancreatitis: a predictor of infection. AB - OBJECTIVE: A retrospective evaluation was performed of the use of gallium imaging in patients with known severe pancreatitis to detect infection in pancreatic and peripancreatic fluid collections. MATERIALS AND METHODS: Gallium-67 single-photon emission computed tomography (SPECT) studies were retrospectively reviewed in patients with complicated pancreatitis. Only patients who had undergone interventional procedures within 10 days of the scanning were included in our analysis. A total of 23 scans from 20 patients were reviewed. SPECT imaging was typically performed 48-72 hr after injection of the gallium. All studies were correlated with conventional CT findings. Findings from subsequent interventions (results of aspiration, Gram stains, or cultures) were used as evidence of infection. RESULTS: Twenty patients underwent either percutaneous or surgical drainage within 10 days of their gallium scanning. One patient underwent gallium scanning on three different occasions and underwent three different interventional procedures after each of the gallium scans, bringing the total number of cases in our study to 23. Of these 23 cases, 18 patients (78%) with gallium scans showing positive findings for infection had infected fluid; five patients (22%) with negative findings for infection on gallium scans had sterile fluid (p < 0.00001). No false-positive scans were found among our study cases, and we found no correlation between the uptake of gallium and the presence or absence of pancreatic necrosis. CONCLUSION: Gallium does not actively accumulate in all patients with severe pancreatitis, and gallium uptake does not correlate with the presence or absence of necrosis. In patients with severe pancreatitis complicated by fluid collections or inflammatory masses, gallium SPECT is a useful predictor of infection and can be used to help guide subsequent intervention. Gallium SPECT allows targeting sites of infected fluid in patients with multiple fluid collections and potentially obviates intervention in patients with sterile fluid collections. PMID- 11906861 TI - Fluoroscopically guided placement of a covered self-expandable metallic stent for malignant antroduodenal obstructions: preliminary results in 18 patients. AB - OBJECTIVE: The purpose of this study was to investigate the technical feasibility and the clinical effectiveness of fluoroscopically guided placement of covered self-expandable metallic stents in the treatment of malignant antroduodenal obstructions. SUBJECTS AND METHODS: With fluoroscopic guidance, covered self expandable metallic stents were placed in 18 consecutive patients with inoperable malignant antroduodenal obstructions. All patients were treated for severe nausea and recurrent vomiting. RESULTS: Stent placement was technically successful in all patients with or without gastrostomy (n = 2) and balloon dilatation (n = 3). After stent placement, symptoms improved in all but one patient, who had another stenosis in the proximal jejunum. During the follow-up of 2-73 weeks (mean, 12 weeks), stent migration occurred in three patients (16.7%) from 1 to 41 days after the procedure. These patients were treated successfully by means of placing a second covered metallic stent. Two patients, who were followed up for longer than 30 weeks, showed a recurrence of strictures because of mechanical failure of the stents; one of the patients was treated with coaxial placement of a second covered metallic stent, which had a positive clinical outcome. CONCLUSION: Fluoroscopically guided placement of covered self-expandable metallic stents is technically feasible and effective for the palliative treatment of inoperable malignant antroduodenal obstructions. The rate of stent migration in our study was lower than those in previous reports. PMID- 11906862 TI - Mesenteric adenitis: CT diagnosis of primary versus secondary causes, incidence, and clinical significance in pediatric and adult patients. AB - OBJECTIVE: Our objective was to determine the clinical significance of mesenteric adentitis when detected on CT. MATERIALS AND METHODS: Mesenteric adenitis was considered present if a cluster of three or more lymph nodes measuring 5 mm or greater each was present in the right lower quadrant mesentery. If no other abnormality was detected on CT, then mesenteric adenitis was considered primary. If a specific inflammatory process was detected in addition to the lymphadenopathy, then mesenteric adenitis was considered secondary. Patients with a known neoplasm or HIV infection were excluded. Three separate groups of patients were examined for the presence and cause of mesenteric adenitis. Group 1 consisted of 60 consecutive patients prospectively identified with mesenteric adenitis on CT examinations. Group 2 consisted of 60 consecutive patients undergoing abdominal and pelvic CT for evaluation of blunt or penetrating abdominal trauma. Group 3 consisted of 60 consecutive patients undergoing abdominal and pelvic CT with acute abdominal symptoms. In all patients, the indication for imaging was documented, and the size of the largest lymph node, when present, was measured. In patients with mesenteric adenitis, the CT findings, clinical history, and clinical or surgical follow-up were subsequently evaluated to determine the cause of mesenteric adenitis. RESULTS: In the 60 patients prospectively identified with CT findings of mesenteric adenitis (group 1), 18 (30%) of 60 had primary mesenteric adenitis. The remaining 42 patients (70%) had an associated inflammatory condition that was established on CT as the likely cause of mesenteric adenitis. Mesenteric adenitis was present in none (0%) of the 60 patients in group 2 and in five (8.3%) of 60 patients in group 3. CONCLUSION: The incidence of mesenteric adenitis in patients with and those without abdominal pain is low. When evidence of mesenteric adenitis is present on CT examinations, usually a specific diagnosis can be established as its cause. PMID- 11906863 TI - Intrathoracic migration of the wrap after laparoscopic Nissen fundoplication: radiologic evaluation. AB - OBJECTIVE: The purpose of our study was to evaluate the different types of postoperative herniation of the wrap into the thorax after laparoscopic Nissen fundoplication, to propose a clear radiologic definition, and to establish their respective frequencies. SUBJECTS AND METHODS: Two hundred twenty-six consecutive patients who underwent laparoscopic Nissen fundoplication were studied prospectively. All patients underwent an upper gastrointestinal series before surgery and on the first postoperative day. Radiologic follow-up performed yearly after surgery in 148 patients (65%) consisted of a double-contrast upper gastrointestinal series. Intrathoracic migration of the wrap was diagnosed on radiography when the intact fundoplication wrap herniated partially or entirely through the esophageal hiatus of the diaphragm. The kappa statistic was used to assess interobserver agreement. RESULTS: Of the 148 upper gastrointestinal series, 44 intrathoracic migrations (30%) were diagnosed. These examinations were reviewed and allowed us to differentiate two types of migrations. Type I (31 patients) consists of a paraesophageal hernia of a portion of the wrap through the esophageal hiatus with the esogastric junction remaining below the diaphragm. Type II (13 patients) is diagnosed when the entire fundoplication herniates through the hiatus with the gastroesophageal junction located at or above the level of the diaphragm. CONCLUSION: Intrathoracic migration is an important complication of laparoscopic Nissen fundoplication. Most migrations are small and asymptomatic. We propose a simple and reproducible radiologic definition of two different types of intrathoracic migration of the wrap observed after laparoscopic Nissen fundoplication. PMID- 11906864 TI - Graded compression sonography with adjuvant use of a posterior manual compression technique in the sonographic diagnosis of acute appendicitis. AB - OBJECTIVE: We evaluated the usefulness of graded compression sonography with the adjuvant use of a posterior manual compression technique for detection of the vermiform appendix and the diagnosis of acute appendicitis. SUBJECTS AND METHODS: Five hundred seventy consecutive patients referred for suspected acute appendicitis were prospectively examined by original, graded compression sonography with a 5- or a 7.5-MHz linear transducer. A posterior manual compression technique was added for 85 patients whose vermiform appendix was not identified with graded compression sonography. For consensus, another experienced radiologist or a resident observer was in attendance throughout the examination. The detection rate for the vermiform appendix and the diagnostic accuracy for acute appendicitis before and after the adjuvant use of a posterior manual compression technique were obtained, respectively, and final diagnoses were established with the official radiology reports, surgical results, and clinical follow-up. RESULTS: Graded compression sonography enabled visualization of the vermiform appendix in 485 (85%) of 570 patients. After the adjuvant use of a posterior manual compression technique, the vermiform appendix was found in an additional 57 of 85 patients, with the number of identified vermiform appendices increasing to 542 (95%) of 570 patients. The 57 patients with an additionally found appendix included 11 patients with acute appendicitis. The sonographic diagnosis of acute appendicitis was determined in 312 of 542 patients. Acute appendicitis was proven by surgery in 311 of 332 patients. Sonography was used to establish the diagnosis in 302 of the 311 patients with proven appendicitis; there were 10 false-positive diagnoses and nine false-negative diagnoses. One false-positive diagnosis was acquired after use of the posterior manual compression technique. These results showed more improvement than those of the probabilities for acute appendicitis with single use of graded compression sonography. CONCLUSION: Graded compression sonography with adjuvant use of a posterior manual compression technique seems to be useful for detecting the vermiform appendix and for diagnosing acute appendicitis. PMID- 11906865 TI - Benign hepatic nodules in Budd-Chiari syndrome: radiologic-pathologic correlation with emphasis on the central scar. AB - OBJECTIVE: The purpose of this study was to determine the imaging features of benign hepatic nodules in patients with Budd-Chiari syndrome and to correlate them with pathologic findings, with special attention placed on the presence of a central scar. MATERIALS AND METHODS: Imaging findings of 59 benign hepatic nodules in four patients with chronic Budd-Chiari syndrome were analyzed retrospectively, and radiologic- pathologic correlation was performed in three patients with 50 hepatic nodules who underwent liver transplantation. All patients underwent multiphasic helical CT. In three patients with 29 lesions, MR imaging, including a multiphasic dynamic study, was performed. The CT and MR imaging findings in these patients were compared with those of 103 small hepatocellular carcinomas in 56 other patients (54 of them displayed chronic hepatitis or liver cirrhosis associated with viral hepatitis but none had Budd Chiari syndrome). Image analysis was performed by two radiologists with no knowledge of the diagnosis. RESULTS: All patients with Budd-Chiari syndrome exhibited multiple benign nodules up to 3 cm in diameter, and 42 of 59 lesions were hypervascular. Microscopically, 15 of 32 nodules demonstrated a central scar; moreover, some nodules closely resembled focal nodular hyperplasia. Frequencies of hyperintensity on T1-weighted images (14/29 vs 25/103), hypointensity on T2-weighted images (7/29 vs 1/103), and the presence of a central scar (6/59 vs 1/103) were significantly higher in benign nodules than in hepatocellular carcinomas (p < 0.05; Fisher's exact test). Moreover, for lesions larger than 1 cm, imaging studies revealed a central scar in six of 15 benign lesions. CONCLUSION: Benign hepatic nodules in patients with in Budd-Chiari syndrome are usually small, multiple, and hypervascular. The presence of a central scar is a characteristic feature in those larger than 1 cm in diameter. PMID- 11906867 TI - Large regenerative nodules in Budd-Chiari syndrome and other vascular disorders of the liver: CT and MR imaging findings with clinicopathologic correlation. AB - OBJECTIVE: The purpose of this study was to determine the CT and MR imaging appearance of large regenerative nodules arising in livers with vascular disorders and to correlate these findings with the clinical and pathologic findings. CONCLUSION: Large regenerative nodules are a characteristic feature of Budd-Chiari syndrome and other hepatic vascular disorders. CT and MR imaging show consistent features of the nodules and the surrounding liver that may allow distinction of Budd-Chiari nodules from other types of hypervascular hyperplastic or dysplastic nodules. PMID- 11906868 TI - Sequential hemodynamic change in hepatocellular carcinoma and dysplastic nodules: CT angiography and pathologic correlation. AB - OBJECTIVE: The purpose of this study was to clarify the hemodynamic changes associated with hepatocarcinogenesis using CT angiography. MATERIALS AND METHODS: Eighty-six hepatocellular lesions were confirmed at pathology in 49 patients who underwent CT with both hepatic arteriography and arterioportography. These images were compared with lesion-to-liver vascular ratios of cumulative arteries, preexisting hepatic arteries, and portal veins in resected specimens. Lesions were classified in five groups according to intranodular hemodynamics determined by CT hepatic arteriography and CT during arterioportography: group 1, isoattenuating on both procedures; group 2, hypoattenuating on CT hepatic arteriography and isoattenuating on CT during arterioportography; group 3, hypoattenuating on both procedures; group 4, isoattenuating on CT hepatic arteriography and hypoattenuating on CT during arterioportography; and group 5, hyperattenuating on CT hepatic arteriography and hypoattenuating on CT during arterioportography. RESULTS: Among 86 lesions, we identified seven low-grade dysplastic nodules, eight high-grade dysplastic nodules, 14 well-differentiated hepatocellular carcinomas, 45 moderately differentiated hepatocellular carcinomas, and 12 poorly differentiated hepatocellular carcinomas. The lesions were classified as group 1 (n = 5), group 2 (n = 13), group 3 (n = 6), group 4 (n = 2), or group 5 (n = 60). Intranodular hemodynamics was significantly correlated with pathologic grading (p < 0.001). For correlations between combinations of the groups and pathologic gradings, the order "groups 1-2-3-4-5" was the most significant (p < 0.001). CONCLUSION: During hepatocarcinogenesis, most hepatocellular nodules show deterioration of arterial blood flow before loss of portal blood flow. Vascular imaging of hepatic nodules may predict malignant abnormality via the early loss of hepatic arterial flow seen before portal flow changes. PMID- 11906869 TI - Small hypervascular hepatocellular carcinoma revealed by double arterial phase CT performed with single breath-hold scanning and automatic bolus tracking. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of double arterial phase CT for the detection of small hypervascular hepatocellular carcinomas, using an automated bolus-tracking technique to initiate the hepatic arterial phase CT. MATERIALS AND METHODS: Double arterial and late phase contrast enhanced helical CT scans were obtained on 287 consecutive patients suspected of having hepatocellular carcinoma. These included 56 patients with 90 small (< or 3 cm) hepatocellular carcinomas and 50 patients with no hepatocellular carcinomas. CT scans of these patients were interpreted by three reviewers. The first arterial phase scan was initiated automatically 10 sec after the bolus-tracking program detected the threshold enhancement of 50 H in the abdominal aorta. Three reviewers interpreted the late phase CT scans in combination with the first, second, or both hepatic arterial phases. Measures of the reviewers' detection of hepatocellular carcinoma included analysis of interobserver variation, sensitivity, specificity, and area under receiver operating characteristic curve (A(z)). RESULTS: The time elapsed from bolus initiation to threshold aortic enhancement ranged from 10 to 24 sec (mean, 13 sec), resulting in initiation of the first arterial phase CT scan from 20 to 34 sec (mean, 23 sec). The combination of late phase CT and both first and second arterial phase images showed significantly better performance than the combination of the late phase and either the first or second arterial phases, although the difference was most evident in comparison with the combination of second arterial and late phases. CONCLUSION: An automated bolus-tracking program can be used to optimize the timing of hepatic arterial phase CT. Multiphasic CT performed using this technique is useful in detection of small hepatocellular carcinoma. PMID- 11906870 TI - Skin injury after radiofrequency ablation for hepatic cancer. PMID- 11906871 TI - Using kinematic MR cholangiopancreatography to evaluate biliary dilatation. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of kinematic MR cholangiopancreatographic (MRCP) images as an aid in predicting the need for intervention in patients with biliary dilatation. SUBJECTS AND METHODS: Fifty patients with suspected pancreaticobiliary disorders were separated into three groups. Group I patients had biliary dilatation with periampullary lesions (n = 16), group II patients had supraampullary obstructive lesions (n = 17), and group III patients had biliary dilatation without obstruction (n = 17). Twenty consecutive single thick-slice MRCP images were obtained in the 15 degrees or 30 degrees left anterior oblique coronal plane. Two radiologists jointly reviewed the images without knowledge of the final diagnosis. The numbers of images showing relaxation of the sphincteric segment and the configuration of the distal margin of the common bile duct for the three groups were compared. RESULTS: Relaxation of the sphincteric segment was observed on the images of only two patients (12%) in group I but on the images of all patients in groups II and III, although not on all images. Lack of visualization of sphincteric relaxation on the kinematic MRCP images had a sensitivity of 88% and a specificity of 100% for the diagnosis of periampullary lesions. Most patients whose images did not show sphincteric relaxation required biliary intervention at the sphincter level. CONCLUSION: Nonvisualization of sphincteric relaxation on kinematic MRCP indicates ampullary or periampullary lesions. Kinematic MRCP can be used to determine the necessity of biliary intervention in patients with biliary dilatation. PMID- 11906872 TI - Directed biopsy during contrast-enhanced sonography of the prostate. AB - OBJECTIVE: We evaluated the value of directed biopsy for the detection of prostate cancer during contrast-enhanced endorectal sonography. SUBJECTS AND METHODS: Forty patients were evaluated with harmonic gray-scale sonography. The evaluation was performed before administration of contrast agent, during continuous IV infusion of perflutren lipid microspheres, and again during bolus administration of the microspheres. Sextant biopsy sites were scored prospectively on a six-point scale for suggestion of malignancy at baseline during contrast infusion and after bolus administration. An additional directed core was obtained at 20 of the sextant biopsy sites based on contrast-enhanced imaging. RESULTS: Cancer was identified in 30 biopsy sites in 16 of the patients (40%). A suspicious site identified during contrast-enhanced transrectal sonography was 3.5 times more likely to have positive biopsy findings at than an adjacent site that was not suggestive of malignancy (p < 0.025). When a suspicious site was evaluated with an additional biopsy core, the site was five times more likely to have a biopsy with positive findings than a standard sextant site (p < 0.01). We found no difference in diagnostic accuracy between continuous infusion of contrast material and bolus administration. CONCLUSION: Contrast enhanced transrectal sonography improves the sonographic detection of malignant foci in the prostate. The performance of multiple biopsies of suspicious enhancing foci significantly improves the detection of cancer. There is no advantage to additional examination of the gland after bolus administration of contrast material. PMID- 11906873 TI - Renal vein Doppler sonography of obstructive uropathy. AB - OBJECTIVE: Obstructive uropathy in the early stages can be difficult to diagnose using either standard sonography or the arterial resistive index. We tested the hypothesis that acute obstruction of the renal collecting system reduces the intraparenchymal renal compliance, which affects the intraparenchymal venous blood flow to a greater degree than the arterial flow. SUBJECTS AND METHODS: Twelve patients with clinical evidence of acute obstructive uropathy were referred for helical CT to confirm the diagnosis and to provide a gold standard by which we could evaluate the sonographic findings in the 12 test patients. Twelve patients without renal disease served as a control group. Doppler sonography of the interlobar arteries and veins of both kidneys then was performed, with the sonographer unaware of which kidney had an obstruction. Peak venous flow measurements and arterial resistive and venous impedance indexes were obtained. The impedance indexes of the obstructed and unobstructed kidney were compared for each patient. RESULTS: The mean arterial resistive indexes of the obstructed kidneys were larger than those of the unobstructed kidneys, 0.67 +/- 0.08 and 0.62 +/- 0.05, respectively (p = 0.05). The venous impedance indexes comparing obstructed and unobstructed sides were 0.38 +/- 0.25 and 0.80 +/- 0.25, respectively, a statistically significant result (p = 0.0002). On average, the peak venous flow signal in the obstructed kidney was 69% higher than that of the unobstructed kidney (p = 0.04) and 86% higher than that of the peak venous flow signal in the control group (p = 0.005). CONCLUSION: Renal obstruction alters the venous flow to a greater extent than the arterial flow, and a comparison between the venous flow in the obstructed and unobstructed kidneys may improve diagnostic accuracy. PMID- 11906874 TI - Clinical importance of a unilateral striated pattern seen on sonography of the testicle. AB - OBJECTIVE: The purpose of this study was to determine the clinical importance of a unilateral striated pattern seen on high-resolution sonography of a testicle on the basis of follow-up imaging. CONCLUSION: In the absence of relevant clinical findings or an abnormal signal on color-flow or power Doppler sonography, a striated pattern of a testicle appears to have no clinical importance. It is presumed to represent fibrosis, and the patient most likely can be followed up clinically and sonographically rather than having to undergo surgical exploration. PMID- 11906875 TI - Fat in renal cell carcinoma that lacks associated calcifications. PMID- 11906876 TI - Metastatic seeding of a percutaneous nephrostomy tract causing cervical carcinoma. PMID- 11906877 TI - Calcification in lymphoma occurring before therapy: CT features and clinical correlation. AB - OBJECTIVE: Calcification in lymphoma occurring before therapy is rare. We assessed the prevalence, CT features, and clinical significance of calcification in nodes and masses in patients with lymphoma occurring before therapy. SUBJECTS AND METHODS: CT of the chest, abdomen, and pelvis of 956 newly diagnosed patients with lymphoma was evaluated prospectively for calcifications in enlarged lymph nodes and lymphoma masses. Findings were correlated with histologic type of disease, tissue parameters, and clinical course. Calcifications were further evaluated on follow-up CT. RESULTS: Of 956 patients with lymphoma (704 with non Hodgkin's lymphoma and 252 with Hodgkin's lymphoma), eight patients (0.84%) showed calcifications in involved sites, seven of whom had non-Hodgkin's lymphoma and one of whom had Hodgkin's lymphoma. Calcifications were present in lymph nodes and masses in the mediastinum in five patients, in the retroperitoneum in two patients, and in the adrenal in one patient. All eight patients had the aggressive type of lymphoma. Four patients later relapsed, one of whom died. A fifth patient died after only minimal response to treatment. CONCLUSION: Calcification in patients with lymphoma occurring before therapy is rare as opposed to that in lymphoma after therapy. It occurred in our patients more often in the mediastinum, in patients with non-Hodgkin's lymphoma rather than in patients with Hodgkin's lymphoma, and only in patients with the aggressive type of disease. PMID- 11906878 TI - Improved image interpretation with registered thoracic CT and positron emission tomography data sets. AB - OBJECTIVE: The purpose of this study was to determine if nonlinear registration of clinically acquired thoracic CT and FDG positron emission tomography (PET) data sets supports more detailed interpretation of metastatic thoracic disease when compared with interpretations from nonregistered PET studies. CONCLUSION: In 11 of 16 data sets of patients imaged for detection of metastatic disease, interpretations from PET studies were correctly altered with registration information. All changes were either improvements in tumor localization or correct interpretation of less metastatic involvement. PMID- 11906879 TI - A closer look at the midsternal stripe sign. AB - OBJECTIVE: Reports in the literature offer conflicting data on the value of the midsternal stripe sign in diagnosing sternal dehiscence. Our purpose was to determine the frequency with which the midsternal stripe sign is present in patients with dehiscence compared with a control group without this complication and to determine whether this sign adds incremental value to the sign of sternal wire displacement in the diagnosis of dehiscence. CONCLUSION: A midsternal stripe thicker than 3 mm should raise one's suspicion of the presence of sternal dehiscence. However, this sign is rarely observed in patients with this complication and does not add incremental value to the finding of sternal wire displacement in establishing the diagnosis of dehiscence. PMID- 11906880 TI - Malignant pleural mesothelioma with osteoblastic heterologous elements: CT and MR imaging findings. PMID- 11906882 TI - Cardiac systolic rotation and contraction before and after valve replacement for aortic stenosis: a myocardial tagging study using MR imaging. AB - OBJECTIVE: Aortic stenosis leads to the derangement of cardiac function and contraction mode because of chronic pressure overload that is relieved after surgical valve replacement. The purpose of this study was to determine the changes in left ventricular systolic rotation and contraction using MR tagging in patients with aortic stenosis before and after surgical valve replacement compared with age-matched healthy volunteers. MATERIALS AND METHODS: Twelve patients with aortic stenosis were examined with an electrocardiographically triggered two-dimensional tagging sequence at 1.5 T before and 12 months after surgical valve replacement for the evaluation of wall function of the apical, mid ventricular, and basal levels. Eight healthy volunteers in the same age group served as the control group. RESULTS: Before surgery, all patients showed a significant increase of apical rotation (22.2 degrees +/- 5.9 degrees vs 10.3 degrees +/- 2.5 degrees, p < 0.0001) and overall left ventricular torsion (25.1 degrees +/- 6.6 degrees vs 14.5 degrees +/- 3.7 degrees, p < 0.001); basal rotation was not significantly different (-2.9 degrees +/- 2.1 degrees vs -4.2 degrees +/- 1.9 degrees, p = not significant) compared with the volunteer group. Apical rotation and torsion were negatively correlated with left ventricular mass (r = -0.73, p < 0.01, and r = -0.61, p < 0.05, respectively) and end-diastolic volume (r = -0.73, p < 0.01 and r = -0.64, p < 0.03, respectively). One year after surgery, basal rotation was reduced in the patients with aortic stenosis compared with the patients in the control group (-1.9 degrees +/- 1.8 degrees, p < 0.01). In comparison with preoperative values, apical rotation (14.2 degrees +/ 3.6 degrees, p < 0.01) also decreased but was still elevated, and this resulted in a normalization of left ventricular torsion (16.1 degrees +/- 3.7 degrees, p < 0.01). CONCLUSION: Surgical valve replacement for aortic stenosis leads to normalization of the left ventricular torsion 1 year after surgery. Pressure overload before surgery is associated with an increase of systolic left ventricular wringing motion, possibly serving as a compensatory mechanism. This mechanism declines with increasing left ventricular hypertrophy and dilatation. PMID- 11906883 TI - Diffusion-weighted echoplanar MR imaging of the salivary glands. AB - OBJECTIVE: We determined the apparent diffusion coefficients of normal and dysfunctional salivary glands. SUBJECTS AND METHODS: A diffusion-weighted single shot spin-echo type of echoplanar MR imaging was performed on the parotid or submandibular glands, or both, in 36 healthy subjects, 20 patients with Sjogren's syndrome, and six patients with sialoadenitis. The apparent diffusion coefficient of the salivary gland was calculated using two b factors (b = 500 and 1,000 sec/mm(2)). RESULTS: The apparent diffusion coefficient was lower in the parotid glands (0.28 x 10(-3) mm(2)/sec) than that of the submandibular glands (0.37 x 10(-3) mm(2)/sec). The apparent diffusion coefficient was increased in sialoadenitis, whereas it decreased with abscess formation. The apparent diffusion coefficients of the parotid glands in patients with Sjogren's syndrome correlated with the salivary flow rates but not with the sialographic gradings of the glands. We also found a correlation of the decreases in apparent diffusion coefficients with the severity of gland damage as assessed on T1-weighted MR images. CONCLUSION: Diffusion-weighted echoplanar MR imaging may reveal diseased states of the salivary glands. PMID- 11906884 TI - Patterns of premature physeal arrest: MR imaging of 111 children. AB - OBJECTIVE: The purpose of this study was to use MR imaging, especially fat suppressed three-dimensional (3D) spoiled gradient-recalled echo sequences, to identify patterns of growth arrest after physeal insult in children. MATERIALS AND METHODS: We evaluated 111 children with physeal bone bridges (median age, 11.4 years) using MR imaging to analyze bridge size, location in physis, signal intensity, growth recovery lines, avascular necrosis, and metaphyseal cartilage tongues. Fifty-eight patients underwent fat-suppressed 3D spoiled gradient recalled echo imaging with physeal mapping. The cause, bone involved, radiographic appearance, and surgical interventions (60/111) were also correlated. Data were analyzed with the two-tailed Fisher's exact test. RESULTS: Posttraumatic bridges, accounting for 70% (78/111) of patients, were most often distal, especially of the tibia (n = 43) and femur (n = 14), whereas those due to the other miscellaneous causes were more frequently proximal (p < 0.0001). The position of the bridge in the physis was related to the bone involved (p < 0.0001). Sixty-five percent of distal tibial bridges involved the anteromedial physis, whereas 60% of the distal femoral arrests were central. Larger bridges had higher T1 signal intensity (p < 0.008). Oblique growth recovery lines were seen exclusively with bridges involving the peripheral physis (p = 0.002) and smaller, more potentially resectable bridges. Metaphyseal cartilaginous tongues were seen with all causes, but avascular necrosis was exclusively posttraumatic (p = 0.03). Signal characteristics and bridge size did not vary with the cause. CONCLUSION: Premature physeal bony bridging in children is most often posttraumatic and disproportionately involves the distal tibia and femur where bridges tend to develop at the sites of earliest physiologic closure, namely anteromedially and centrally, respectively. MR imaging, especially with the use of fat-suppressed 3D spoiled gradient-recalled echo imaging, exquisitely shows the growth disturbance and associated abnormalities that may follow physeal injury and guides surgical management. PMID- 11906885 TI - Femoral head osteochondral lesions in painful hips of athletes: MR imaging findings. AB - OBJECTIVE: This study describes the MR imaging findings of focal osteochondral lesions found in the hips of 11 athletes with persistent pain and normal findings on radiographs. CONCLUSION: Osteochondral lesions of the femoral head are seen on MR imaging as focal, medial areas of high T2-weighted and low T1-weighted signals and should be considered as a possible cause of persistent hip or groin pain in young, high-level athletes because the institution of appropriate treatment may help to prevent late degenerative sequelae. PMID- 11906886 TI - The "reverse Segond" fracture: association with a tear of the posterior cruciate ligament and medial meniscus. AB - OBJECTIVE: We describe three patients who presented with radiographic findings of a fragment on the medial side of the tibial plateau of the knee that represented an avulsion of the deep portion of the medial collateral ligament. These findings were all associated with disruption of the posterior cruciate ligament and a peripheral medial meniscal tear-the so-called reverse Segond fracture. CONCLUSION: Avulsion fracture at the tibial insertion of the deep component of the medial collateral ligament is a rare finding. When this type of injury is diagnosed, the radiologist should consider posterior cruciate ligament injury and peripheral medial meniscal tears as possible associated findings. PMID- 11906887 TI - Skeletal muscle metastases at sites of documented trauma. AB - OBJECTIVE: Hematogenous metastases to skeletal muscles have been reported to be rare. We report eight biopsy-proven cases of skeletal metastases occurring in sites of previously documented skeletal muscle trauma. We retrospectively reviewed MR imaging examinations obtained at a large orthopedic surgical oncology service from January 1994 through December 2000 for biopsy-proven metastases to skeletal muscles. Our retrospective review revealed 28 patients with biopsy proven skeletal muscle metastases. Of these 28 patients, eight had a documented clinical history of previous trauma at the site of skeletal metastasis. Five of these eight patients underwent MR imaging before the development of a metastasis. MR imaging revealed a hematoma in three of the five patients and a partial muscle tear in two of the five patients. The hematomas and partial muscle tears were in the same skeletal muscle location in which the metastatic disease subsequently developed. Metastatic disease was documented by MR imaging and subsequent biopsy. CONCLUSION: Skeletal muscle injury may alter muscle physiology and result in increased susceptibility to the development of metastatic disease at such sites. PMID- 11906888 TI - Dermatofibrosarcoma protuberans: MR imaging features. AB - OBJECTIVE: The objective of our study was to describe the MR imaging features of 10 cases of histologically confirmed dermatofibrosarcoma protuberans. CONCLUSION: MR imaging is useful in identifying the extent and location of dermatofibrosarcoma protuberans. Although most cases of this tumor are superficial and well defined, we have shown three cases in which the tumor was in a deep location and one case in which the tumor was ill defined in appearance. Knowledge of the variable MR imaging appearances of these tumors may aid in the diagnosis of difficult or atypical cases. PMID- 11906889 TI - Hoffa fracture: a common association with high-energy supracondylar fractures of the distal femur. PMID- 11906890 TI - Vector analysis of the wall shear rate at the human aortoiliac bifurcation using cine MR velocity mapping. AB - OBJECTIVE: Small or oscillatory wall shear stress accelerates atherosclerosis. MR velocity mapping is feasible for vector analysis of wall shear rate (a spatial gradient of blood flow velocity at the vessel wall) in humans. A relationship between anatomic variations at the aortoiliac bifurcation and characteristics of wall shear rate was evaluated. SUBJECTS AND METHODS: To obtain two components of wall shear rate vectors, an axial component along the vessel axis and a nonaxial component perpendicular to the former at the inner and outer walls of the common iliac arteries just distal to the aortoiliac bifurcation, we performed cine MR velocity mapping with three orthogonal velocity-encoded directions in seven volunteers. RESULTS: The peak axial component at the outer wall (120.6 +/- 37.2 sec(-1)) was smaller than that at the inner wall (196.0 +/- 53.7 sec(-1)) (p < 0.01). Oscillation described by a time integral of the axial component in recessive blood flow direction over integrals in dominant and recessive directions at the outer wall was greater (0.24 +/- 0.11) than that at the inner wall (0.15 +/- 0.08) (p < 0.01). The intersecting angle between the extrapolation of the aortic axis and the direction of the axis of the common iliac artery correlated positively with the peak axial component (r = 0.577, p < 0.05) and inversely with oscillation (r = 0.603, p < 0.05). CONCLUSION: Three-dimensional vector analysis with MR velocity mapping revealed that the outer wall at the aortoiliac bifurcation showed low and oscillatory shear rate, and this inclination was increased when the takeoff angle of the iliac artery was small. PMID- 11906891 TI - Deep vein thrombosis: can a second sonographic examination be avoided? AB - OBJECTIVE: Our objective was to determine the prevalence of deep vein thrombosis in symptomatic patients and its distribution based on the assessment of prior clinical probability. We evaluated whether repeated sonography is necessary in patients with either intermediate or high clinical probability for deep vein thrombosis after an initial examination with negative findings. SUBJECTS AND METHODS: We prospectively evaluated 438 consecutive patients with clinical suggestion of deep vein thrombosis of the lower limbs, classified according to the prior clinical probability (high, intermediate, low). Sonography with positive findings was diagnostic for deep vein thrombosis. Negative findings in low-risk patients excluded thrombosis. Patients with intermediate or high clinical risk whose initial sonographic examination showed negative findings underwent a second examination after 1 week. RESULTS: Of the 438 patients with clinical symptoms, 112 patients (26%) had positive findings on sonography, and 326 (74%) had negative findings. Of the 202 intermediate- and high-risk patients with negative initial sonography, 140 patients underwent a single follow-up sonographic examination 1 week later. In three cases, findings were positive for deep vein thrombosis. Two other patients developed pulmonary embolism. Sonographic follow-up increased the detection of deep vein thrombosis in the patients with intermediate or high probability from 32.5% to 33.5%; the prevalence of thromboembolic disease in this group was 34%. CONCLUSION: The prevalence of deep vein thrombosis studied by sonography in the patients with intermediate or high clinical risk was 33.5%. Initial sonography revealed a 32.5% prevalence, and a second examination 1 week later detected an additional 1%. Sonography did not reveal 0.5% of thromboembolic events. Our results do not justify a routine second scanning at 1 week. PMID- 11906892 TI - Sonographically guided biopsy of suspicious microcalcifications of the breast: a pilot study. AB - OBJECTIVE: The purpose of this study is to evaluate the use of sonographic guidance for biopsy of mammographically detected suspicious microcalcifications. SUBJECTS AND METHODS: Twenty-three patients with suspicious microcalcifications detected on mammography (15 associated with masses or distortion; eight with microcalcifications alone) underwent sonographically guided core biopsy (n = 18) or sonographically guided needle localization before excision (n = 5). Microcalcifications were targeted, and specimen radiographs were obtained for each lesion, with the success of the procedure based on identifying microcalcifications on the specimen radiograph. For core biopsies, the number of cores obtained was compared with that in 49 control patients who underwent sonographically guided core biopsy of noncalcified masses. RESULTS: All 23 lesions (100%) were successfully biopsied under sonographic guidance, with microcalcifications seen on specimen radiographs in each case. Of 18 core biopsies, a mean of 8.7 cores was obtained compared with a mean of 5.5 cores in the control group (p<0.0001). Of 13 lesions sampled with core biopsy that subsequently underwent surgical excision, three (23%) were upgraded from atypical ductal hyperplasia to ductal carcinoma in situ (n = 1) and from ductal carcinoma in situ to invasive carcinoma (n = 2). Mammographically, most lesions contained more than 15 pleomorphic microcalcifications. On sonography, echogenic foci corresponded to microcalcifications in all but two cases in which broader echogenic regions were seen. When no mass or distortion was visible on mammography, sonography showed a mass or dilated ducts with internal echogenic foci. CONCLUSION: Microcalcifications identifiable on sonography can be successfully biopsied under sonographic guidance. Further study is necessary to determine whether targeting microcalcifications seen sonographically in the mass or duct can improve the rate of underestimation of disease compared with stereotactic core biopsy. PMID- 11906893 TI - Stereotactic biopsy of the breast using an upright unit, a vacuum-suction needle, and a lateral arm-support system. AB - OBJECTIVE: This study evaluated the vacuum-suction needle (8- to 11-gauge) on an upright stereotactic machine with a lateral arm-support system. SUBJECTS AND METHODS: From July 1999 to August 2000, 185 core biopsies of the breast in 179 consecutive patients were planned in four institutions using 8- 11-gauge vacuum suction probes on an upright stereotactic unit. Needles were stabilized and attached to the x, y, and z coordinates of the machines via a lateral arm-support system. Needle entry was made in the x-axis. RESULTS: Five patients were canceled, and 180 biopsies were performed in 174 patients while the patients were in seated (n = 171) and lateral decubitus (n = 9) positions. An average of 9.5 cores were taken (range, 5-26 cores). Targeting was successful in 176 (98%) of 180 cores. Lesions were missed because of movement associated with Parkinson's disease (n = 1), or because the mass was obscured (n = 1) or calcifications were not in the core specimen (n = 2). Findings in 152 (84%) of the biopsies were benign and 28 (16%) were malignant. Forty-one lesions underwent surgical excision and 106 underwent mammographic follow-up. Discordance was 4% (6/147). Complications included vasovagal reactions (n = 10, 5.6%), bleeding (n = 5, 3%), hematomas (n = 3, 1.7%), vomiting (n = 1, 0.6%), and technical failure (n = 1, 0.6%). CONCLUSION: Vacuum-suction needle core biopsies can be successfully performed on an upright stereotactic machine with a lateral arm attachment. Thinly compressed breasts and lesions located near the chest wall are well sampled. The vasovagal rate is higher than that on a prone table but is acceptable. PMID- 11906894 TI - Sonographic evaluation of subcutaneous hemangioma of the breast. PMID- 11906895 TI - Embolization for the treatment of adenomyosis. PMID- 11906896 TI - Full disclosure of breast biopsy options. PMID- 11906897 TI - Clinical value of positron emission tomography. PMID- 11906898 TI - CT of small-bowel obstruction. PMID- 11906899 TI - Immediate reporting of screening mammography. PMID- 11906900 TI - On the passing of Harold Jacobson. PMID- 11906902 TI - Lysophosphatidic acid and its role in reproduction. AB - Lysophosphatidic acid (LPA) belongs to a new family of lipid mediators that are endogenous growth factors and that elicit diverse biological effects, usually via the activation of G protein-coupled receptors. LPA can be generated after cell activation through the hydrolysis of preexisting phospholipids in the membranes of stimulated cells. A dramatic elevation of LPA levels was found in serum of patients suffering from ovarian carcinoma. Because these high LPA amounts can be detected as early as stage I of the disease, LPA has been introduced as a new marker for ovarian cancer. Progression of the malignancy is correlated with a differential expression of various LPA receptor subtypes. The presence of LPA in the follicular fluid of healthy individuals implicates that this biological mediator may be relevant to normal ovarian physiology. LPA induces proliferation and mitogenic signaling of prostate cancer cells, and a novel LPA receptor isoform has been recognized in healthy prostate tissues. This evidence indicates multiple roles for LPA in both male and female reproductive physiology and pathology. In this review, we summarize the literature on LPA generation, the way it is degraded, and the mechanisms by which signals are transduced by various LPA receptors in reproductive tissues, and we discuss possible future research directions in these areas. PMID- 11906903 TI - Subcellular distribution of ZP1, ZP2, and ZP3 glycoproteins during folliculogenesis and demonstration of their topographical disposition within the zona matrix of mouse ovarian oocytes. AB - The zona pellucida (ZP) is an extracellular coat synthesized and secreted by the oocyte during follicular development and surrounding the plasma membrane of mammalian eggs. To date, the mechanism of synthesis and secretion, mode of assembly, and intracellular trafficking of the ZP glycoproteins have not been fully elucidated. Using antibodies against mouse ZP1, ZP2, and ZP3 in conjunction with the protein A-gold technique, we have shown an association of immunolabeling with the Golgi apparatus, secretory granules, and a complex structure called vesicular aggregate, respectively, in mouse ovarian follicles. In contrast, the neighboring granulosa cells were not reactive to any of the three antibodies used. Immunolabeling of ZP1, ZP2, and ZP3 was detected throughout the entire thickness of the ZP, irrespective of the developmental stage of ovarian follicles. Double and triple immunolocalization studies, using antibodies tagged directly to different sizes of gold particles, revealed an asymmetric spatial distribution of the three ZP glycoproteins in the zona matrix at various stages of follicular development. All three glycoproteins were specifically localized over small patches of darkly stained flocculent substance dispersed throughout the zona matrix. Very often, ZP1, ZP2, and ZP3 were found in close association. These results confirm findings from previous studies demonstrating that ovarian oocytes and not granulosa cells are the only source for mouse ZP glycoproteins. In addition, results from our morphological and immunocytochemical experiments suggest that the vesicular aggregates in the ooplasm are likely to serve as an intermediary in the synthesis and secretion of ZP glycoproteins. The stoichiometric disposition of ZP1, ZP2, and ZP3 in the zona matrix as revealed by double and triple immunolocalization studies provide further insight into some of the unanswered questions pertinent to the current model of mouse ZP structure proposed by the Wassarman group. PMID- 11906904 TI - Dynamics of testicular germ cell proliferation in normal mice and transgenic mice overexpressing rat androgen-binding protein: a flow cytometric evaluation. AB - Transgenic mice carrying rat androgen-binding protein (ABP) genomic DNA express high amounts of testicular ABP and develop a progressive impairment of spermatogenesis. To understand the mechanism of these changes, we have studied the pattern of testicular germ cell proliferation from 7 to 360 days of age in wild-type (WT) control and transgenic homozygous (ABP-TG) mice by flow cytometry after labeling DNA in isolated germ cells with propidium iodide. At all ages studied, the body weight of the ABP-TG mice was lower than that of age-matched WT controls. Significantly reduced testicular weight and total germ cell number in the ABP-TG mice were evident from Day 30 and Day 60, respectively. Flow cytometric analysis of isolated germ cells revealed that the number of germ cells undergoing proliferation (S-phase cells) was identical in WT control and ABP-TG mice up to Day 14. Subsequently, the number of germ cells in S-phase was consistently higher in ABP-TG than in WT mice. The number of primary spermatocytes was significantly increased starting from Day 60, and the numbers of round and elongated spermatids were significantly reduced in the ABP-TG animals from Day 21 and Day 60 onwards, respectively. Immunocytometry for intracellular ABP at 90 days of age revealed that the percentage of ABP containing germ cells was greater in ABP-TG than in WT mice. The continuous presence of ABP in mouse seminiferous tubules at greater than physiological concentrations facilitates the formation of primary spermatocytes but impairs subsequent transformation to round and elongated spermatids. Based on our observations and the analysis of the available literature, the most likely mechanism for production of these effects is sustained reduction in the bioavailability of androgens. PMID- 11906905 TI - Efficacy and safety of a new vaginal contraceptive antimicrobial formulation containing high molecular weight poly(sodium 4-styrenesulfonate). AB - Host cell infection by sexually transmitted disease (STD)-causing microbes and fertilization by spermatozoa may have some mechanisms in common. If so, certain noncytotoxic agents could inhibit the functional activity of both organisms. High molecular mass poly(sodium 4-styrenesulfonate) (T-PSS) may be one of these compounds. T-PSS alone (1 mg/ml) or in a gel (2% or 5% T-PSS) completely prevented conception in the rabbit. Contraception was not due to sperm cytotoxicity or to an effect on sperm migration. However, T-PSS inhibited sperm hyaluronidase (IC(50) = 5.3 microg/ml) and acrosin (IC(50) = 0.3 microg/ml) and caused the loss of acrosomes from spermatozoa (85% maximal loss by 0.5 microg/ml). T-PSS (5% in gel) also reduced sperm penetration into bovine cervical mucus (73% inhibition by 1 mg gel/ml). T-PSS (5% in gel) inhibited human immunodeficiency virus (HIV; IC(50)= 16 microg gel/ml) and herpes simplex viruses (HSV-1 and HSV-2; IC(50) = 1.3 and 1.0 microg gel/ml, respectively). The drug showed high efficacy against a number of clinical isolates and laboratory strains. T-PSS (5% in gel) also inhibited Neisseria gonorrhea (IC(50) < 1.0 gel/ml) and Chlamydia trachomatis (IC(50) = 1.2 microg gel/ml) but had no effect on lactobacilli. These results imply that T-PSS is an effective functional inhibitor of both spermatozoa and certain STD-causing microbes. The noncytotoxic nature should make T-PSS safe for vaginal use. T-PSS was nonmutagenic in vitro and possessed an acute oral toxicity of >5 g/kg (rat). Gel with 10% T-PSS did not irritate the skin or penile mucosa (rabbit) and caused no dermal sensitization (guinea pig). Vaginal administration of the 5% T-PSS gel to the rabbit for 14 consecutive days caused no systemic toxicity and only mild (acceptable) vaginal irritation. T-PSS in gel form is worthy of clinical evaluation as a vaginal contraceptive HIV/STD preventative. PMID- 11906906 TI - Enhanced survivability of cloned calves derived from roscovitine-treated adult somatic cells. AB - Nuclear transfer to produce cattle is inefficient because 1) donor cells are not easily synchronized in the proper phase of the cell cycle, 2) the nucleus of these cells is not effectively reprogrammed, 3) the rate of attrition of late term pregnancies is high, and 4) the health of early postnatal calves is compromised. The cyclin dependent kinase 2 inhibitor, roscovitine, was used to maximize cell cycle synchrony and to produce cells that responded more reliably to nuclear reprogramming. Roscovitine-treated adult bovine granulosa cells (82.4%) were more efficiently synchronized (P < 0.05) in the quiescent G0/G1 phase of the cell cycle than were serum-starved cells (76.7%). Although blastocyst development following nuclear transfer was elevated (P < 0.05) in the serum-starved group (21.1%) relative to the roscovitine-treated cells (11.8%), the number of cells in the blastocysts derived from roscovitine-treated cells was higher (P < 0.05) than those derived from the serum-starved group (roscovitine treated group: 142.8 +/- 6.0 cells; serum-starved group: 86.8 +/- 14.5 cells). The resulting fetal and calf survival after embryo transfer was enhanced in the roscovitine-treated group (seven surviving calves from six pregnancies) compared with serum-starved controls (two calves born, one surviving beyond 60 days, from five pregnancies). Roscovitine culture can predictably synchronize the donor cell cycle and increase the nuclear reprogramming capacity of the cells, resulting in enhanced fetal and calf survival and increased cloning efficiency. PMID- 11906907 TI - Effect of the absence or presence of various protein supplements on further development of bovine oocytes during in vitro maturation. AB - The evaluation of culture medium for bovine oocytes has progressed toward more defined conditions during the last few years. The main objective of this study was to evaluate different sources of albumin as a protein supplement during in vitro maturation (IVM) of bovine oocytes in synthetic oviduct fluid medium (SOF). The replacement of protein with polyvinyl pyrrolidone (PVP) or polyvinyl alcohol (PVA) was also evaluated. The effect of recombinant human FSH on cumulus expansion and nuclear maturation in SOF containing BSA (BSA-V) or PVP-40 was also studied. Addition of BSA-V during IVM retarded nuclear maturation when compared with addition of PVP-40 or use of SOF alone. The inclusion of different concentrations of BSA-V, fetal calf serum (FCS), or PVA during IVM had no positive effect on the developmental capacity of the oocytes compared with the use of SOF alone with no supplement but significantly decreased the percentage of embryos reaching the morula and blastocyst stages. However, when BSA-V was replaced with purified BSA, BSA that was essentially free of fatty acids, or chicken egg albumin, embryonic development rates were restored. The presence of PVP-40 but not PVP-360 during IVM significantly increased morula and blastocyst production. These results indicate that although SOF alone can support bovine oocyte maturation, a high proportion of morulae and blastocysts can be produced from IVM oocytes cultured in medium containing PVP-40. These studies are the first to show that the effect of FSH on nuclear maturation and cumulus expansion is dependent on substrates present in IVM medium. PMID- 11906908 TI - Onset of steroidogenic enzyme gene expression during ovarian follicular development in sheep. AB - Steroidogenesis is a major function of the developing follicle. However, little is known about the stage of onset of steroid regulatory proteins during follicular development in sheep. In this study, several steroidogenic enzymes were studied by immunohistochemistry and/or in situ hybridization; cytochrome P450 side chain cleavage (P450(scc)), cytochrome P450 17alpha-hydroxylase (17alphaOH), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), cytochrome P450 aromatase (P450(arom)), steroidogenic factor 1 (SF-1), steroidogenic acute regulatory protein (StAR), and LH receptor (LH-R). To define the stages of follicular growth, ovarian maps were drawn from serial sections of ovine ovaries, and follicles were located and classified at specific stages of growth based on morphological criteria. In this way, the precise onset of gene expression with respect to stages of follicular growth for all these proteins could be observed. The key findings were that ovine oocytes express StAR mRNA at all stages of follicular development and that granulosa cells in follicle types 1-3 express 3beta-HSD and SF-1. Furthermore, the onset of expression in theca cells of StAR, P450(scc), 17alphaOH, 3beta-HSD, and LH-R occurred in large type 4 follicles just before antrum formation. This finding suggests that although the theca interna forms from the type 2 stage, it does not become steroidogenically active until later in development. These studies also confirm that granulosa cells of large type 5 follicles express SF-1, StAR, P450(scc), LH-R, and P450(arom) genes. These findings raise new questions regarding the roles of steroidogenic regulatory factors in early follicular development. PMID- 11906909 TI - Smad 3 may regulate follicular growth in the mouse ovary. AB - Although Smad 3 is known to serve as a signaling intermediate for the transforming growth factor beta (TGFbeta) family in nonreproductive tissues, its role in the ovary is unknown. Thus, we used a recently generated Smad 3-deficient (Smad 3-/-) mouse model to test the hypothesis that Smad 3 alters female fertility and regulates the growth of ovarian follicles from the primordial stage to the antral stage. In addition, we tested whether Smad 3 affects the levels of proteins that control apoptosis, survival, and proliferation in the ovarian follicle. To test this hypothesis, breeding studies were conducted using Smad 3-/ and wild-type mice. In addition, ovaries were collected from Smad 3-/- and wild type mice on Postnatal Days 2-90. One ovary from each animal was used to estimate the total number of primordial, primary, and antral follicles. The other ovary was used for immunohistochemical analysis of selected members of the B-cell lymphoma/leukemia-2 family of protooncogenes (Bax, Bcl-2, Bcl-x), proliferating cell nuclear antigen (PCNA), and cyclin-dependent kinase 2 (Cdk-2). The results indicate that Smad 3-/- mice have reduced fertility compared with wild type mice. The results also indicate that Smad 3 may not affect the size of the primordial follicle pool at birth, but it may regulate growth of primordial follicles to the antral stage. Further, the results indicate that Smad 3 may regulate the expression of Bax and Bcl-2, but not Bcl-x, Cdk-2, and PCNA. Collectively, these data suggest that Smad 3 may play an important role in the regulation of ovarian follicle growth and female fertility. PMID- 11906910 TI - Fetal programming: prenatal androgen disrupts positive feedback actions of estradiol but does not affect timing of puberty in female sheep. AB - We studied the impact of prenatal androgen exposure on the timing of onset of puberty, maintenance of cyclicity in the first breeding season, and the LH surge mechanism in female sheep. Pregnant sheep were injected with testosterone propionate (100 mg i.m.) twice each week from Day 30 to Day 90 (D30-90) or from Day 60 to Day 90 (D60-90) of gestation (term = 147 days). Concentrations of plasma progesterone and gonadotropins were measured in blood samples collected twice each week from control (n = 10), D60-90 (n = 13), and D30-90 (n = 3) animals. Rate of weight gain and initiation of estrous behavior were also monitored. After the first breeding season, when the animals entered anestrus, competency of the gonadotropin surge system to respond to estradiol positive feedback was tested in the absence or presence of progesterone priming for 12 days. Prenatally androgenized females had similar body weight gain and achieved puberty (start of first progestogenic cycle) at the same time as controls. Duration of the breeding season and the number of cycles that occurred during the first breeding season were similar between control and prenatally androgenized sheep. In contrast, prenatal exposure to androgens compromised the positive feedback effects of estradiol. Onset of LH/FSH surges following the estradiol stimulus was delayed in both groups of androgenized ewes compared with the controls in both the absence and presence of progesterone priming. In addition, the magnitude of LH and FSH surges in the two animals that surged in the D30-90 group were only one third and one half, respectively, of the magnitudes observed in the control and D60-90 groups. The present findings indicate that disruption of the surge system can account for the fertility problems that occur during adulthood in prenatally androgenized sheep. PMID- 11906911 TI - Dynamic changes in the expression of relaxin-like factor (INSL3), cholesterol side-chain cleavage cytochrome p450, and 3beta-hydroxysteroid dehydrogenase in bovine ovarian follicles during growth and atresia. AB - Relaxin-like factor (RLF) is a new member of the insulin-relaxin gene family known to be expressed in the ovarian follicular thecal cells of ruminants. To investigate the pattern of RLF expression in development and atresia of bovine follicles, antisera were raised in rats and rabbits to recombinantly expressed bovine pro-RLF and to chemically synthesized ovine RLF B chain, respectively. On dot blotting analysis, the rat antiserum bound to pro-RLF and less strongly to a synthetic mature ovine RLF lacking the C-domain, whereas the rabbit antiserum bound the mature form of ovine RLF. These antisera were used to immunostain bovine ovarian follicles of differing sizes and stages of health and atresia. 3beta-Hydroxysteroid dehydrogenase was colocalized with pro-RLF (n = 86 follicles), and cholesterol side-chain cleavage cytochrome P450 was localized in another section of many of the same follicles (n = 66). Not all follicles expressed pro-RLF in the theca interna, so the results are presented as the proportion of follicles expressing pro-RLF. Both mature and pro-RLF were immunolocalized to steroidogenic thecal cells of healthy follicles. As follicles enlarged to >5 mm, the proportion expressing pro-RLF declined (19/19 for <5 mm and 18/26 for >6 mm). Atresia was divided into antral (antral granulosa cells dying first) or basal (basal cells dying first) and further divided into early, middle, and late. For antral atresia of small follicles (2-5 mm), no decline in the proportion expressing pro-RLF was observed (early 6/6, middle 2/2) until the late stages (1/4). For basal atresia, which only occurs in small follicles (2-5 mm), the proportion expressing pro-RLF declined in the middle (2/5) and late (0/8) stages. In larger follicles (>6 to <10 mm), the proportion expressing pro RLF also declined with atresia (1/13). These declines in RLF expression with atresia or increasing size were not accompanied by a decline in the expression of steroidogenic enzymes in the theca interna. A significant (P < 0.001) inverse relationship in the expression of pro-RLF and 3beta-hydroxysteroid dehydrogenase in the membrana granulosa was observed. We conclude that the expression of pro RLF in the theca interna is switched off as follicles enlarge or enter atresia, whereas the expression of steroidogenic enzymes is maintained in the theca interna. PMID- 11906912 TI - Functional analysis of stem cells in the adult rat testis. AB - Adult stem cells maintain several self-renewing systems and processes in the body, including the epidermis, hematopoiesis, intestinal epithelium, and spermatogenesis. However, studies on adult stem cells are hampered by their low numbers, lack of information about morphologic or biochemical characteristics, and absence of functional assays, except for hematopoietic and spermatogonial stem cells. We took advantage of the recently developed spermatogonial transplantation technique to analyze germ line stem cells of the rat testis. The results indicate that the stem cell concentration in rat testes is 9.5-fold higher than that in mouse testes, and spermatogenic colonies derived from rat donor testis cells are 2.75 times larger than mouse-derived colonies by 3 mo after transplantation. Therefore, the extent of spermatogenesis from rat stem cells was 26-fold greater than that from mouse stem cells at the time of recipient testis analysis. Attempts to enrich spermatogonial stem cells in rat testis populations using the experimental cryptorchid procedure were not successful, but selection by attachment to laminin-coated plates resulted in 8.5 fold enrichment. Spermatogonial stem cells are unique among adult stem cells because they pass genetic information to the next generation. The high concentration of stem cells in the rat testis and the rapid expansion of spermatogenesis after transplantation will facilitate studies on stem cell biology and the introduction of genetic modifications into the male germ line. The functional differences between spermatogonial stem cells of rat vs. mouse origin after transplantation suggest that the potential of these cells may vary greatly among species. PMID- 11906913 TI - Bax-dependent spermatogonia apoptosis is required for testicular development and spermatogenesis. AB - Bax is a multidomain, proapoptotic member of the Bcl-2 family that is required for normal spermatogenesis in mice. Despite its proapoptotic function, previous results found that Bax-deficient mature male mice demonstrate increased cell death and dramatic testicular atrophy. The present study examined the role of Bax during the normal development of the testis to determine whether the increased cell death in mature mice could be explained by decreased apoptosis earlier in development. Consistent with this hypothesis, testicular atrophy is preceded by increased testicular weight and hypercellular tubules in immature Bax-deficient mice. TUNEL staining at Postnatal Day (P) 7 and morphological quantitation between P5 and P15 demonstrates decreased germ cell apoptosis in Bax-deficient mice. By P15, increased numbers of type A spermatogonia, and at P12 and P15, an increase in intermediate type spermatogonia were noted in Bax-deficient animals. By P25, the number of basal compartment cells was greatly increased in Bax deficient animals compared with controls such that four or five layers of preleptotene spermatocytes were routinely present within the basal compartment of the testis. Although the Sertoli cell barrier was significantly removed from the basement membrane, it appeared intact as judged by the hypertonic fixation test. During late pubertal development, massive degeneration of germ cells took place, including many of those cell types that previously survived in the first wave of spermatogenesis. The data indicate that Bax is required for normal developmental germ cell death in the type A spermatogonia, specifically dividing (A(2), A(3), and A(4)) spermatogonia, at a time at which the number of spermatogonia is regulated in a density-dependent manner. The massive hyperplasia that occurs in Bax-deficient mice subsequently results in Bax independent cell death that may be triggered by overcrowding of the seminiferous epithelium. PMID- 11906914 TI - Effects of progestins on progesterone synthesis in a stable porcine granulosa cell line: control of transcriptional activity of the cytochrome p450 side-chain cleavage gene. AB - The purpose of the present study was to examine the effects of progestins on progesterone synthesis and expression of the cytochrome P450 cholesterol side chain cleavage gene (P450(scc)) in a stable porcine granulosa cell line, the JC 410. Cells were incubated for 48 h with the synthetic progestogen-levornorgestrel with or without RU486 (progesterone and glucocorticoid receptor antagonist) or RWJ26819 (progesterone agonist without affinity to glucocorticoid receptors). Both levonorgestrel and RU486 enhanced progesterone accumulation in a dose dependent manner. RU486 did not antagonize the effects of levonorgestrel, and RWJ26819 had no effect on progesterone production in cultured JC-410 cells. Progesterone and levonorgestrel increased steady state P450(scc) mRNA levels after 3-6 h of treatment. Progesterone and RU486 at 0.1, 1, and 10 microM increased the transcription rate of P450(scc) transiently expressed in JC-410 cells after 18 h of incubation; 30 microM had no effect, and 100 microM suppressed transcription. Levonorgestrel did not affect transcription of the P450(scc) gene, and RWJ26819 reduced its transcription. Progesterone and RU486 significantly decreased the number of cells and total protein content after 72 and 24 h of incubation, respectively. Levonorgestrel had no effect, whereas RWJ26819 increased (24 h) but subsequently reduced (72 h) cell number and protein content. The present results indicate that progestins are capable of directly modulating progesterone biosynthesis in porcine JC-410 granulosa cells. These effects may be exerted in part through the regulation of P450(scc) gene expression. Ostensible differences exist between progesterone and its synthetic analogues in the control of progesterone secretion in the stable porcine granulosa cell line in vitro. PMID- 11906915 TI - Changes in Leydig cell gene expression during development in the mouse. AB - Developmental changes in the expression of 18 Leydig cell-specific mRNA species were measured by real-time polymerase chain reaction to partially characterize the developmental phenotype of the cells in the mouse and to identify markers of adult Leydig cell differentiation. Testicular interstitial webs were isolated from mice between birth and adulthood. Five developmental patterns of gene expression were observed. Group 1 contained mRNA species encoding P450 side chain cleavage (P450(scc)), P450(c17), relaxin-like factor (RLF), glutathione S transferase 5-5 (GST5-5), StAR protein, LH receptor, and epoxide hydrolase (EH); group 2 contained 3beta-hydroxysteroid dehydrogenase (3beta-HSD) VI, 17beta hydroxysteroid dehydrogenase (17beta-HSD) III, vascular cell adhesion molecule 1, estrogen sulfotransferase, and prostaglandin D (PGD)-synthetase; group 3 contained patched and thrombospondin 2 (TSP2); group 4 contained 5alpha-reductase 1 and 3alpha-hydroxysteroid dehydrogenase; group 5 contained sulfonylurea receptor 2 and 3beta-HSD I. Group 1 contained genes that were expressed in fetal and adult Leydig cells and which increased in expression around puberty toward a maximum in the adult. Group 2 contained genes expressed only in the adult Leydig cell population. Group 3 contained genes with predominant fetal/neonatal expression in the interstitial tissue. Group 4 contained genes with a peak of expression around puberty, whereas genes in group 5 show little developmental change in expression. Highest mRNA levels in descending order were RLF, P450(c17), EH, 17beta-HSD III, PGD-synthetase, GST5-5, and P450(scc). Results identify five genes expressed in the mouse adult Leydig cell population, but not in the fetal population, and one gene (TSP2) that may be expressed only in the fetal Leydig cell population. The developmental pattern of gene expression suggests that three distinct phases of adult Leydig cell differentiation occur. PMID- 11906916 TI - Occurrence of estrogen receptor alpha in bovine placentomes throughout mid and late gestation and at parturition. AB - The bovine placenta produces estrogens from the first trimester until the end of its life span. However, with the exception of the immediate prepartal and intrapartal phases, in which an involvement of placental estrogens has been suggested for the preparation of parturition, their function has not been elucidated yet. To test for a role of placental estrogens as local factors regulating placental growth and differentiation, placentomes from cows that were pregnant for 150, 220, 240, and 270 days, and parturient cows (3 animals per group) were screened immunohistochemically for the expression of estrogen receptor alpha (ERalpha). Indirect immunoperoxidase staining methods were applied using primary monoclonal antibodies (pmAbs) directed against the C-terminus (AER311, HT277) or the N-terminus (AER314, 1D5) of the ERalpha molecule. Both types of pmAbs identified ERalpha in stromal cells and capillary pericytes of the maternal caruncular septae. Using pmAb 1D5, the mean percentage of ERalpha positive caruncular stromal cells decreased from 39.0% +/- 5.9% in pregnant cows to 17.5% +/- 8.3% at parturition (P = 0.011). Only pmAb recognizing the C terminus identified ERalpha in the caruncular epithelium, in which positive reactions were found in all cells, with the exception of areas adjacent to the chorionic plate and to major chorionic villi, where the specific signal gradually faded and occasionally disappeared. No positive reactions were observed in the fetal part of the placentomes. The expression of ERalpha in bovine placentomes was further confirmed by the detection of ERalpha-specific mRNA by reverse transcriptase-polymerase chain reaction and by Western blot analysis. The results suggest a role for placental estrogens as paracrine factors involved in the regulation of placental growth and differentiation. PMID- 11906917 TI - Dynamic testicular adhesion junctions are immunologically unique. I. Localization of p120 catenin in rat testis. AB - In the seminiferous epithelium, morphologically diverse junctions mediate inter Sertoli and Sertoli-germ cell adhesive contact, but the molecular composition of such junctions is not well known. At prototypical adherens junctions, proteins termed catenins bind to the intracellular domain of classic cadherins and regulate the strength of adhesion. Using a panel of monoclonal antibodies (5A7, 8D11, and 15D2), p120 catenin (p120) was localized in postnatal and adult rat testis cryosections and touch preparations by immunofluorescence. Immunoprecipitation of testis homogenates showed that at least four p120 isoforms were expressed from Postnatal Day 7 through adulthood. Both inter-Sertoli and Sertoli-germ cell junctions were p120-positive, however, individual p120 monoclonals were localized to specific junctions. The 5A7 and 8D11 antibodies colocalized with beta-catenin and plectin at inter-Sertoli and Sertoli spermatocyte junctions. At inter-Sertoli junctions, p120 was juxtaposed to but did not colocalize with f-actin. Thus, p120 is likely a component of inter Sertoli desmosome-like junctions. In contrast, the 15D2 monoclonal antibody specifically immunostained Sertoli-round spermatid and inter-Sertoli cell junctions in a dynamic pattern. From the time that round spermatids form to their differentiation into elongate spermatids, Sertoli-round spermatid 15D2 immunostaining cycled from a single mass to a curvilinear pattern, and finally to punctate structures scattered throughout the epithelium. This localization and stage-specific immunostaining pattern indicated that 15D2 recognized Sertoli round spermatid desmosome-like junctions. Between Sertoli cells, 15D2 immunostained newly formed junctions (at Postnatal Days 21 through 43), but not mature junctions in the adult. From these data, we conclude that p120 is a component of most, if not all, desmosome-like junctions, and that desmosome-like junctions between different cell types contain a unique molecular composition. PMID- 11906918 TI - Dynamic testicular adhesion junctions are immunologically unique. II. Localization of classic cadherins in rat testis. AB - In the seminiferous epithelium, morphologically diverse junctions mediate inter Sertoli and Sertoli-germ cell adhesive contact and likely transmit signals between contacting cells. Defining the molecular composition of testicular cell cell junctions is an important step in determining their function. Proteins belonging to the cadherin superfamily are important mediators of cell-cell adhesion, as well as cell signaling. Here, we determined the spatial and temporal protein expression of four classic cadherins in rat testis: N-cadherin, cadherin 6, cadherin-11, and a cadherin defined by an antiserum generated against a conserved classic cadherin peptide (L4). Through Western blot analysis, all antibodies recognized unique proteins. Similarly, each cadherin displayed unique, cell-type specific immunostaining patterns. Whereas N-cadherin, cadherin-11, and L4-positive cadherin were expressed from Postnatal Day 7 through adulthood, cadherin-6 protein was not present at Postnatal Day 7 and first appeared at Day 21. Immunostaining of testis cryosections on Postnatal Days 7, 21, 31, 43, and those of adults indicated that cadherin-11 localized to peritubular cell junctions. N-cadherin immunostaining localized to basal inter-Sertoli junctions, Sertoli-spermatocyte junctions, and at about stages I-VII in Sertoli-elongate spermatid junctions. Cadherin-6 immunostaining was restricted to Sertoli-round spermatid and in Sertoli-elongate spermatid junctions at approximately stages XII I. Finally, L4-positive immunostaining also detected Sertoli-round spermatid junctions in addition to Sertoli-elongate spermatid junctions at approximately stages XII-I. These data show that the various testicular cell-cell junctions are molecularly unique and dynamic complexes. PMID- 11906919 TI - Mosaic gene expression in nuclear transfer-derived embryos and the production of cloned transgenic pigs from ear-derived fibroblasts. AB - Genetically modified domestic animals have many potential applications ranging from basic research to production agriculture. One of the goals in transgenic animal production schemes is to reliably predict the expression pattern of the foreign gene. Establishing a method to screen genetically modified embryos for transgene expression before transfer to surrogates may improve the likelihood of producing offspring with the desired expression pattern. In order to determine how transgene expression may be regulated in the early embryo, we generated porcine embryos from two distinct genetically modified cell lines by using the nuclear transfer (NT) technique. Both cell lines expressed the enhanced green fluorescent protein (eGFP); the first was a fibroblast cell line derived from the skin of a newborn pig that expressed eGFP, whereas the second was a fetal derived fibroblast cell line into which the eGFP gene was introduced by a retroviral vector. The reconstructed embryos were activated by electrical pulses and cultured in NCSU23. Although the in vitro developmental ability of each group of NT embryos was not different, the eGFP expression pattern was different. All embryos produced from the transduced fetal cell line fluoresced, but only 26% of the embryos generated from the newborn cell line fluoresced, and among those that did express eGFP, more than half had a mosaic expression pattern. This was unexpected because the fetal cell line was not clonally selected, and each cell had potentially different sites of integration. Embryos generated from the newborn cell line were surgically transferred to five surrogate gilts. One gilt delivered four female piglets, all of which expressed eGFP, and all had microsatellites identical to the donor. Here we demonstrate that transgene expression in all the blastomeres of an NT embryo is not uniform. In addition, transgene expression in a genetically manipulated embryo may not be an accurate indicator of expression in the resulting offspring. PMID- 11906920 TI - Stage-specific expression of the intermediate filament protein cytokeratin 13 in luminal epithelial cells of secretory phase human endometrium and peri implantation stage rabbit endometrium. AB - In preparation for blastocyst implantation, uterine luminal epithelial cells express new cell adhesion molecules on their apical plasma membrane. Since one mechanism epithelial cells employ to regulate membrane polarity is the establishment of specific membrane-cytoskeletal interactions, this study was undertaken to determine if new cytokeratin (CK) intermediate filament assemblies are expressed in endometrial epithelial cells during developmental stages related to blastocyst implantation. Type-specific CK antibodies were used for immunocytochemical and immunoblot analyses of 1) intermediate filament networks of the endometrial epithelium during embryo implantation in rabbits and 2) proliferative and secretory phases of the human menstrual cycle. CK18, a type I CK found in most simple epithelia, was expressed in all luminal and glandular epithelial cells of both the human and rabbit endometrium at all developmental stages analyzed; it was also strongly expressed in trophectoderm of the implanting rabbit blastocyst. In contrast, CK13, another type I cytokeratin, exhibited a regulated expression pattern in luminal, but not glandular, epithelial cells of secretory phase human and peri-implantation stage rabbit endometrium. Furthermore, in the rabbit implantation chambers, CK13 was predominantly localized at the cell apex of luminal epithelial cells, where it assembled into a dense filamentous network. These data suggest that the stage specific expression of CK13 and a reorganization of the apical intermediate filament cytoskeleton of uterine luminal epithelial cells may play important functions in preparation for the implantation process. PMID- 11906921 TI - Steroids modulate transepithelial resistance and Na(+) absorption across cultured porcine vas deferens epithelia. AB - Epithelial cells were isolated from adult porcine vas deferens and grown in the absence or presence of steroid hormones. Transepithelial resistance (R(te)), basal short circuit current (I(sc)), and the effects of selected ion transport modulators on these parameters were evaluated in modified Ussing chambers at three time points (5-8, 11-14, and 18-22 days postseeding). At the earliest time point, no significant differences were observed. At the middle time point, when compared with R(te) in untreated control monolayers, R(te) in monolayers exposed to 17beta-estradiol, aldosterone, cortisol, cortisone, prednisolone, prednisone, and dexamethasone was significantly lower; in contrast, R(te) in monolayers exposed to testosterone, dihydrotestosterone, or progesterone did not differ from that in control monolayers. Treatments with cortisol, prednisolone, and dexamethasone were associated with an elevated basal I(sc) that was amiloride sensitive, indicating ongoing Na(+) absorption by these monolayers. R(te) was increased by amiloride treatment in glucocorticoid-treated monolayers but remained significantly less than that of control monolayers. At the third time point, the postamiloride R(te) of glucocorticoid-treated monolayers was not different from that of control monolayers. Responses to ATP, forskolin, bumetanide, and DASU-02 were not affected by steroid treatment at any time point. Taken together, these results suggest that estrogens and corticosteroids can modulate epithelial function in the distal excurrent duct of the adult male reproductive system. At physiological or pharmacological concentrations, these hormones would be expected to modify the luminal environment (both the ionic composition and pH) to which sperm are exposed and thus affect male fertility. PMID- 11906922 TI - Chronic cyclophosphamide treatment alters the expression of stress response genes in rat male germ cells. AB - Increases in the survival rate of men treated with chemotherapeutic drugs and their desire to have children precipitate concerns about the effects of these drugs on germ cells. Azoospermia, oligospermia, and infertility are common outcomes resulting from treatment with cyclophosphamide, an alkylating agent. Exposure of male rats to cyclophosphamide results in dose-dependent and time specific adverse effects on progeny outcome. Elucidation of the effects of chronic low-dose cyclophosphamide treatment on the expression of stress response genes in male germ cells may provide insight into the mechanisms underlying such adverse effects. Male rats were gavaged with saline or cyclophosphamide (6 mg/kg) for 4-5 wk; pachytene spermatocytes, round spermatids, and elongating spermatids were isolated; RNA was extracted and probed on cDNA arrays containing 216 cDNAs. After saline treatment, 125 stress response genes were expressed in pachytene spermatocytes (57% of genes studied), 122 in round spermatids (56%), and 83 in elongating spermatids (38%). Cyclophosphamide treatment reduced the number of genes detected in all germ cell types. The predominant effect of chronic cyclophosphamide exposure was to decrease the expression level of genes in pachytene spermatocytes (34% of genes studied), round spermatids (29%), and elongating spermatids (4%). In elongating spermatids only, drug treatment increased the expression of 8% of the genes studied. The expression profiles of genes involved in DNA repair, posttranslational modification, and antioxidant defense in male germ cells were altered by chronic cyclophosphamide treatment. We hypothesize that the effects of cyclophosphamide exposure on germ cell gene expression during spermatogenesis may have adverse consequences on male fertility and progeny outcome. PMID- 11906923 TI - Successful piglet production after transfer of blastocysts produced by a modified in vitro system. AB - Porcine in vitro production (IVP) systems, including in vitro maturation (IVM) and in vitro fertilization (IVF) of oocytes and their subsequent in vitro culture (IVC), have been modified by many researchers, but are still at a low level because of a low developmental rate of embryos to the blastocyst stage and their poor qualities. Our objectives were to establish reliable IVP procedures for porcine blastocysts and to examine the ability of the blastocysts to develop to term after transfer to recipients. Porcine cumulus-oocyte complexes were matured in vitro under 5% O(2) or 20% O(2), fertilized in vitro under 5% O(2), and subsequently cultured under 5% O(2) in 1) IVC medium supplemented with glucose (IVC-Glu) from Day 0 (the day of IVF) to Day 6; 2) IVC-Glu from Days 0 to 2, then IVC medium supplemented with pyruvate and lactate (IVC-PyrLac) from Days 2 to 6; 3) IVC-PyrLac from Days 0 to 2, then IVC-Glu from Days 2 to 6; and 4) IVC-PyrLac from Days 0 to 6. There were no significant differences in blastocyst formation rates on Day 6 between the 5% O(2) and 20% O(2) conditions (19.9% and 14.0%, respectively). However, the quality of blastocysts, as evaluated by the total cell number, was better after IVM under 5% O(2) than under 20% O(2) (mean cell number, 43.5 and 37.8, respectively). When IVP embryos were cultured in IVC PyrLac from Days 0 to 2 and subsequently in IVC-Glu from Days 2 to 6, the rate of blastocyst formation (25.3%) and cell number (48.7) were higher than the rates (5.8% to 18.1%) and numbers (35.4 to 37.1) with the IVC-Glu then IVC-Glu, the IVC Glu then IVC-PyrLac, and the IVC-PyrLac then IVC-PyrLac regimens, respectively. We then prepared conditioned medium (CM) from culture of porcine oviductal epithelial cells for 2 days in IVC-PyrLac and evaluated its effect on development to the blastocyst stage. Cultivation in CM for the first 2 days, followed by IVC Glu for a further 4 days, had a significantly greater effect in increasing the number of cells in the blastocyst (58.3) than did in IVC-PyrLac (48.4). Finally, we evaluated the ability of blastocysts, generated by IVM under 5% O(2) and IVC in CM, to develop to term. When Day 5 expanding blastocysts (mean cell number, 49.7) were transferred to an estrus-synchronized recipient (50 blastocysts per recipient), the recipient remained pregnant and farrowed eight normal piglets. Furthermore, when Day 6 expanded blastocysts (mean cell number, 80.2) were transferred to two estrus-synchronized recipients, both gilts remained pregnant and farrowed a total of 11 piglets. These results suggest that an excellent piglet production system can be established by using this modified IVP system, which produces high-quality porcine blastocysts. This system has advantages for the generation of cloned and transgenic pigs. PMID- 11906924 TI - Heat shock-initiated apoptosis is accelerated and removal of damaged cells is delayed in the testis of clusterin/ApoJ knock-out mice. AB - The secretion and localization of clusterin in the testis has led to the hypothesis that clusterin plays a role in spermatogenesis. Furthermore, the association of clusterin with apoptosis, cellular injury, disease, and regression of nongonadal tissues has led to the hypothesis that clusterin acts to protect cells from apoptosis or may be involved in tissue remodeling. To investigate the role of clusterin in the testis, we analyzed clusterin knock-out (cluKO) mice to determine the impact of the absence of clusterin on spermatogenesis. Furthermore, we investigated the cellular response to injury caused by methoxyacetic acid (MAA) toxicity and mild heat exposure in the cluKO mice to determine the extent to which clusterin protects against apoptosis or participates in tissue remodeling. We found that cluKO mice were fertile and had essentially normal spermatogenesis with the exception of some incomplete spermiation after stage VIII. No differences in testicular morphology or the incidence of apoptosis in the testis were seen between the cluKO and clusterin wild-type (cluWT) mice after MAA treatment. In contrast, apoptosis was delayed in the cluWT mice compared with the cluKO mice after heat exposure, suggesting that clusterin does have a slight protective effect against apoptosis under some conditions. Also, a dramatic loss of germ cells after heat stress occurred earlier in the cluWT testes than in the cluKO testes. Clusterin is clearly acting in a dual role in that cells can be protected from damage and dead cells can be more easily removed after some types of cellular damage but not after others. PMID- 11906925 TI - Expression of estrogen receptors alpha and beta in the baboon fetal ovary. AB - In adult mammals, estrogen regulates ovarian function, and estrogen receptor (ER) is expressed in granulosa cells of antral follicles of the adult baboon ovary. Because the foundation of adult ovarian function is established in utero, the present study determined whether ERalpha and/or ERbeta were expressed in fetal ovaries obtained on Days 100 (n = 3) and 165-181 (n = 5) of baboon gestation (term = Day 184). On Day 100, ERalpha protein was detected by immunocytochemistry in surface epithelium and mesenchymal-epithelial cells but not oocytes in germ cell cords. ERbeta protein was also detected by immunocytochemistry on Day 100 of gestation and was abundantly expressed in mesenchymal-epithelial cells in germ cell cords, lightly expressed in the germ cells, but was not detected in the surface epithelium. On Days 165-180 of gestation, ERalpha expression was still intense in the surface epithelium, in mesenchymal-epithelial cells throughout the cortex, and in nests of cells between follicles. ERalpha expression was lighter in granulosa cells and was not observed in all granulosa cells, particularly in follicles close to the cortex. In contrast, ERbeta expression was most intense in granulosa cells, especially in flattened granulosa cells, was weaker in mesenchymal-epithelial cells and nests of cells between follicles, and was absent in the surface epithelium. Using an antibody to the carboxy terminal of human ERbeta, ERbeta protein was also detected by Western immunoblot with molecular sizes of 55 and 63 kDa on Day 100 and primarily 55 kDa on Day 180. The mRNAs for ERalpha and ERbeta were also detected by Northern blot analysis in the baboon fetal ovary. These results are the first to establish that the ERalpha and ERbeta mRNAs and proteins are expressed and exhibit changes in localization in the primate fetal ovary between mid and late gestation. Because placental estrogen production and secretion into the baboon fetus increases markedly during advancing pregnancy, we propose that estrogen plays an integral role in programming fetal ovarian development in the primate. PMID- 11906926 TI - Nature of DNA damage in ejaculated human spermatozoa and the possible involvement of apoptosis. AB - Numerous studies have shown the presence of DNA strand breaks in human ejaculated spermatozoa. The nature of this nuclear anomaly and its relationship to patient etiology is however poorly understood. The aim of this study was to investigate the relationship between nuclear DNA damage, assessed using the TUNEL assay and a number of key apoptotic markers, including Fas, Bcl-x, and p53, in ejaculated human spermatozoa from men with normal and abnormal semen parameters. We also determined the nature of the DNA damage by examining the percentage of ejaculated spermatozoa exhibiting DNA damage using the comet assay and by challenging sperm chromatin to attack by micrococcal nuclease S7 and DNase I. We show that TUNEL positivity and apoptotic markers do not always exist in unison; however, semen samples that had a low sperm concentration and poor morphology were more likely to show high levels of TUNEL positivity and Fas and p53 expression. In addition, the DNA damage in ejaculated human sperm is represented by both single- and double-stranded DNA breaks, and access to the DNA is restricted by the compacted nature of ejaculated spermatozoa. This DNA protection is poorer in men with abnormal semen parameters. We propose that the presence of DNA damage is not directly linked to an apoptotic process occurring in spermatozoa and arises due to problems in the nuclear remodeling process. Subsequently, the presence of apoptotic proteins in ejaculated spermatozoa may be linked to defects in cytoplasmic remodeling during the later stages of spermatogenesis. PMID- 11906927 TI - Interval between preovulatory surges of luteinizing hormone increases late in the reproductive period in turkey hens. AB - In turkey hens, the egg production rate is relatively high early during a reproductive period, but declines as the period progresses. Among lines with different egg production potential, the interval between preovulatory surges of LH is the primary determinant of the egg production rate. The main objective of this study was to determine whether the decline in egg production rate late during an egg production period is also associated with a difference in the interval between LH preovulatory surges. A group of photosensitive turkey hens (Early) were photostimulated with continuous light (24L:0D) at 40 wk of age to induce egg laying, and serial blood samples were collected after about 3 wk of egg production. A second group of hens (Late) were housed in floor pens and photostimulated with 14L:10D at 40 wk of age for a normal 36-wk reproduction period and were then switched to 24L:0D lighting for 2 wk before collection of serial blood samples. Continuous light photostimulation was used for at least 2 wk before and during serial blood sampling to avoid potential masking effects of diurnal lighting on the interval between LH surges. The Early (n = 12) and Late (n = 16) hens were cannulated 3 days before being serially bled hourly for 10 days. The mean interval between preovulatory surges of LH was shorter in the Early hens than in the Late hens (26.1 +/- 2.5 h and 34.7 +/- 3.9 h, respectively). The intra-hen LH surge interval coefficient of variation was lower in the Early hens than in the Late hens (7.2% and 18.6%, respectively). The inter hen LH surge interval coefficient of variation was similar in the Early and Late hens (9.5% and 11.2%, respectively). The incidence of blind surges of LH (those not retrospectively associated with ovipositions) was not different between Early and Late laying hens (8.4% +/- 15.2% and 7.3% +/- 14.6%, respectively). In conclusion, in turkey hens, longer intervals and greater intra-hen variation between LH surges were associated with a poorer rate of egg production late in the reproductive period relative to early in the reproductive period. PMID- 11906929 TI - Matrix metalloproteinase (2, 9, and 14) expression, localization, and activity in ovine corpora lutea throughout the estrous cycle. AB - Members of the matrix metalloproteinase (MMP) family collectively degrade extracellular matrix (ECM) and help regulate luteal function. The objectives of these experiments were to characterize the mRNA expression, localization, and activity of MMPs 2, 9, and 14 in ovine corpora lutea (CL). Ovine CL were collected on Days 2, 4, 10, and 15 of the estrous cycle (Day 0 = estrus). Messenger RNA transcripts for MMPs 2 and 14 were detected using Northern analysis; however, expression of MMP-9 was undetectable. Expression of MMP-14 mRNA (membrane type-1 MMP) was increased (P < 0.05) on Day 4; whereas, expression of MMP-2 mRNA was highest (P < 0.05) on Day 10, which corresponded to the observed increases in gelatinolytic activity in luteal homogenates as measured by a fluroscein-labeled gelatin substrate assay. MMP 2 and 9 proteins were localized predominantly to large luteal cells (LLCs), whereas MMP-14 was localized primarily to cells other than LLCs as demonstrated by immunohistochemistry. Immunolocalization of MMP-2 to putative endothelial cells was also observed on Day 15. Localization of MMP activity was determined using in situ zymography. Luteal tissues contained gelatinolytic activity primarily localized pericellularly to various cell types, including LLCs. These results support the hypothesis that ECM remodeling occurs throughout the luteal phase and may help potentiate cellular migration, differentiation, angiogenesis, and growth factor bioavailability. PMID- 11906928 TI - Hepatic lipase deficiency attenuates mouse ovarian progesterone production leading to decreased ovulation and reduced litter size. AB - The lipolytic enzyme hepatic lipase (HL) may facilitate mobilization of cholesterol substrate for ovarian steroidogenesis. We investigated whether HL was necessary for optimum reproduction in the female mouse by analyzing breeding performance and ovarian responses to gonadotropins in HL-/- mice. HL-/- female mice bred with HL-/- males had the same pregnancy success rate and pup survival rate as did wild-type (WT) mice but had significantly smaller litters, producing 1.7 fewer pups per litter. Mice were primed with eCG/hCG, and at 6 h post-hCG the HL-/- mice had smaller ovaries than did the WT mice. HL deficiency specifically affected ovarian weight; adrenal gland weights did not differ between WT and HL-/ mice. HL-/- mice weighed more than age-matched WT mice. Between the two mouse genotypes, uterine weights were the same, indicating that estrogen production was equivalent. However, the HL-/- ovaries produced significantly less progesterone than did the WT ovaries within 6 h of hCG stimulation. HL-/- ovaries had the same number of large antral follicles as did the WT ovaries but had fewer hemorrhagic sites, which represent ovulations, fewer corpora lutea, and more oocytes trapped in corpora lutea. We suggest that reduced progesterone synthesis following hCG stimulation attenuated the final maturation of preovulatory follicles, resulting in smaller ovaries. Furthermore, reduced progesterone production limited the expression of proteolytic enzymes needed for tissue remodeling, resulting in fewer ovulations with a corresponding increase in trapped or unovulated oocytes and providing a possible explanation for the smaller litter size observed in spontaneously ovulating HL-/- mice. PMID- 11906930 TI - Porcine sperm factor supports activation and development of bovine nuclear transfer embryos. AB - A study was undertaken to determine whether injection of porcine sperm factors (pSF), which trigger oscillations in intracellular calcium concentration ([Ca(2+)](i)) in mammalian oocytes, could be used to activate bovine oocytes during nuclear transfer. To date, only combined treatments that induce a monotonic rise in [Ca(2+)](i) and inhibit either phosphorylation or protein synthesis have been utilized in nuclear transfer. Several doses of pSF were tested. Injection of 5 mg/ml pSF triggered [Ca(2+)](i) oscillations that resembled those associated with fertilization with respect to amplitude and periodicity, and as a result, a high percentage of oocytes underwent activation. Furthermore, this concentration of pSF supported in vitro and in vivo development up to 60-90 days of gestation, comparable to development in control nuclear transfer embryos. Nevertheless, neither activation procedure supported development as well as did fertilization. The effectiveness of pSF as an activating agent in bovine oocytes may have been compromised because pSF was unable to support oscillations past 3-5 h postinjection and a second injection was necessary to extend the [Ca(2+)](i) oscillations. Likewise, a single injection of pSF failed to trigger downregulation of the inositol 1,4,5 trisphosphate receptor 1 subtype, whereas a second injection downregulated the receptor in a manner similar to that seen in fertilized oocytes. These results demonstrate that soluble factor(s) from porcine sperm can support early development in bovine nuclear transfer embryos; however, the efficacy may be limited because of the premature cessation of the induced oscillations. PMID- 11906931 TI - Loss of luteinizing hormone surges induced by chronic estradiol is associated with decreased activation of gonadotropin-releasing hormone neurons. AB - Chronic exposure of young ovariectomized rats to elevated circulating estradiol causes loss of steroid-induced LH surges. Such LH surges are associated with cFos induced activation of GnRH neurons; therefore, we hypothesized that chronic estradiol treatment abolishes LH surges by decreasing activation of GnRH neurons. Regularly cycling rats were ovariectomized and immediately received an estradiol implant or remained untreated. Three days or 2 or 4 wk later, the estradiol treated rats received vehicle or progesterone at 1200 h, and 7 hourly blood samples were collected for RIA of LH. Thereafter, all rats were perfused, and the brains were examined for immunocytochemical localization of cFos and GnRH. The GnRH neurons from untreated ovariectomized rats rarely expressed cFos. As reported, LH surges induced by 3 days of estradiol treatment were associated with a 30% increase in cFos-containing GnRH neurons, and progesterone enhanced both the amplitude of LH surges and the proportion of cFos-immunopositive GnRH neurons. As hypothesized, the abolition of LH surges caused by 2 or more weeks of estradiol was paralleled by a reduction in the percentage of cFos-containing GnRH neurons, and this effect was delayed by progesterone. These results suggest that chronic estradiol abolishes steroid-induced LH surges in part by inactivating GnRH neurons. PMID- 11906932 TI - Changes in concentrations of follicular fluid factors during follicle selection in mares. AB - The temporal relationships in the changes in concentrations of follicular fluid factors during follicle selection were characterized in mares. All follicles > or =5 mm were ablated 10 days after ovulation, followed by follicular fluid collection from the three largest follicles (F1, F2, and F3) when F1 of the new wave reached a diameter of 8.0-11.9, 12.0-15.9, 16.0-19.9, 20.0-23.9, 24.0-27.9, or 28.0-31.9 mm (n = 4-8 mares/range). Diameter deviation between F1 and F2 began during the 20.0- to 23.9-mm range, as indicated by a greater difference in diameter between the two follicles at the 24.0- to 27.9-mm range than at the 20.0 to 23.9-mm range. Androstenedione concentrations increased in F1, F2, and F3 between the 16.0- to 19.9- and 20.0- to 23.9-mm ranges. In contrast, estradiol, free insulin-like growth factor (IGF)-1, activin-A, and inhibin-A concentrations increased only in F1 beginning at the 16.0- to 19.9-mm range. As a result, the concentrations of all four factors were higher in F1 than in F2 and F3 at all the later ranges, including the 20.0- to 23.9-mm range (beginning of diameter deviation). Concentrations of progesterone differentially increased in F1, concentrations of androstenedione and IGF-binding protein (IGFBP)-2 increased only in F2 and F3, and concentrations of inhibin-B differentially decreased in F2 and F3 simultaneous with the beginning of deviation. Concentrations of FSH, LH, pro-alphaC inhibin, and total inhibin did not change differentially among follicles. Results indicated that, on a temporal basis, estradiol, free IGF-1, activin-A, and inhibin-A may have played a role in the initiation of follicle deviation. In addition, these four factors as well as progesterone, androstenedione, IGFBP-2, and inhibin-B may have been involved in the subsequent differential development of the follicles. PMID- 11906933 TI - Dynamics of messenger RNAs encoding inhibin/activin subunits and follistatin in the ovary during the rat estrous cycle. AB - Quantitative changes in ovarian inhibin/activin subunit and follistatin mRNAs during the rat estrous cycle were examined by ribonuclease protection assay using digoxygenin-labeled RNA probes. Levels of ovarian inhibin alpha subunit mRNA remained low throughout estrus, metestrus, and diestrus; abruptly increased on the morning of proestrus; then rapidly decreased when the primary gonadotropin surge occurred. A similar changing pattern was observed in inhibin/activin beta(A) subunit mRNA. On the other hand, inhibin/activin beta(B) subunit mRNA showed a different changing pattern. Levels of beta(B) subunit mRNA remained constant during metestrus and diestrus, abruptly decreased on the afternoon of proestrus, then quickly recovered from the nadir by 1100 h on estrus. Throughout the rat estrous cycle, especially during the periovulatory period, alpha subunit mRNA levels were considerably higher than beta(A) and beta(B) subunit mRNA levels. In addition, changes in plasma concentrations of inhibin A and inhibin B were very similar to that in ovarian beta(A) and beta(B) subunit mRNA levels, respectively, with several-hour delays. These results suggest that levels of beta subunit mRNAs restrict secretion of dimeric inhibins. Levels of follistatin mRNA remained low from the midnight of metestrus to the midnight of diestrus, then increased until initiation of the primary gonadotropin surge. Thereafter, follistatin mRNA decreased, reached the nadir at 0200 h on estrus, then increased abruptly at 1100 h on estrus. Afterward, follistatin mRNA levels remained high until the morning of metestrus. The changing pattern of ovarian follistatin mRNA was similar to, and preceded, the changes in plasma concentrations of progesterone, suggesting that ovarian follistatin may modulate progesterone secretion during the rat estrous cycle. PMID- 11906934 TI - Cloning and expression of zebra finch (Taeniopygia guttata) steroidogenic factor 1: overlap with hypothalamic but not with telencephalic aromatase. AB - The zebra finch (Taeniopygia guttata) brain is highly sexually dimorphic. The organization and production of sex-specific song is considerably influenced by estrogens and androgens. Because the brain itself expresses several steroidogenic enzymes, the local production of sex steroids may contribute to sex differences in neural development. Sex steroid production in gonads is directed by a master regulatory factor, steroidogenic factor 1 (SF1). We have identified a cDNA encoding the homologue of SF1 in the zebra finch and utilized reverse transcription-polymerase chain reaction and in situ hybridization to examine early and late developmental expression of SF1 in brain and in early gonadal development. We found that SF1 is expressed early in embryonic development in the Rathke pouch, beginning at stage 15 and extending to at least stage 27 in both males and females. The earliest expression of SF1 in gonads was found at stage 17 for both males and females and extended to at least stage 27. In brain, we assessed SF1 mRNA expression in posthatch and adult telencephalon, and we compared SF1 and aromatase mRNA expression in adult hypothalamus. In the telencephalon and hippocampus, aromatase was expressed independently of SF1, whereas in the hypothalamus, aromatase and SF1 expression were more closely associated. Expression of SF1 and of aromatase overlapped in restricted regions of the hypothalamus, suggesting that SF1 may regulate aromatase expression in these regions. These findings suggest that steroidogenesis in the zebra finch brain may be regulated by both SF1-dependent and SF1-independent mechanisms. No sex differences were detected in SF1 expression in brain. PMID- 11906935 TI - Formation of ovarian follicles during fetal development in sheep. AB - The origin of follicle (i.e., pregranulosa) cells that become the somatic component of primordial follicles is obscure. In addition, information regarding the structural changes that accompany the concomitant regression of ovigerous cords and the appearance of primordial follicles is lacking. In the present study, ovine ovaries collected at frequent time intervals between Day 38 and Day 100 of fetal life were examined by light and electron microscopy. To gain new information regarding the origin of follicular cells, incorporation of 5-bromo-2' deoxyuridine was used to identify proliferating cells at selected stages of development. Based on the location and identity of proliferating cells, apoptotic cells, and sequential changes in histoarchitecture, we hypothesize 1) that most (i.e., >95%) of the granulosal cells in newly formed primordial follicles originate from the ovarian surface epithelium; 2) that the sequential events leading to follicle formation take place entirely within ovigerous cords, with the first follicles forming at the interface of the cortex and medulla; and 3) that the loss (i.e., >75%) of germ cells, but not of somatic cells, within the ovigerous cords is a means by which each surviving oocyte gains additional pregranulosal cells before follicle formation. Conceptual models detailing the chronology of developmental events involved in the formation of primordial follicles in sheep are discussed. PMID- 11906936 TI - Ultrastructure of the resting ovarian follicle pool in healthy young women. AB - In humans, follicle quantity and quality decline with age by atresia. In the present study we aimed to describe the quality of the follicle pool through an ultrastructural investigation of resting follicles in young healthy women. From ovarian biopsies of 7 women aged 25-32 yr, 182 small follicles were morphometrically assessed for various signs of atresia. Morphometric variables were analyzed by principal components analysis (PCA) to demonstrate correlations between variables and to construct an objective follicle score. One third of small follicles consisted of primordial follicles. Nucleus:cell ratios remained constant for oocytes and granulosa cells from primordial to primary follicles, suggesting that follicles up to primary stages belong to the resting pool. The distribution of follicle quality scores as derived from PCA showed that most follicles were of good quality and with little signs of atresia. Atresia in resting follicles appears to be a necrotic process, starting in the ooplasma. Early atresia was characterized by increasing numbers of multivesicular bodies and lipid droplets, dilation of smooth endoplasmic reticulum and Golgi, and irregular mitochondria with changed matrix density. In progressive atresia mitochondrial membranes ruptured, oocyte nuclear membranes were indented or ruptured, and the ooplasma showed extensive vacuolarization. The early involvement of mitochondria in this process suggests that damage is induced by oxygen radicals. PCA follicle quality scores can be reliably approximated using a reduced number of seven morphometric variables, which were selected by stepwise forward analysis. The algorithm to calculate these follicle scores is presented. PMID- 11906937 TI - Differential expression of estrogen receptors alpha and beta in the reproductive tracts of adult male dogs and cats. AB - Expression of estrogen receptors (ERs) in the reproductive tracts of adult male dogs and cats has not been reported. In the present study, ERalpha and ERbeta were localized by immunohistochemistry using ER-specific antibodies. ERalpha was found in interstitial cells and peritubular myoid cells in the dog testis, but only in interstitial cells of the cat. In rete testis of the dog, epithelial cells were positive for ERalpha staining, but in the cat, rete testis epithelium was only weakly positive. In efferent ductules of the dog, both ciliated and nonciliated cells stained intensely positive. In the cat, ciliated epithelial cells were less stained than nonciliated epithelial cells. Epithelial cells in dog epididymis and vas deferens were negative for ERalpha. In the cat, except for the initial region of caput epididymis, ERalpha staining was positive in the epithelial cells of epididymis and vas deferens. Multiple cell types of dog and cat testes stained positive for ERbeta. In rete testis and efferent ductules, epithelial cells were weakly positive for ERbeta. Most epithelial cells of the epididymis and vas deferens exhibited a strong positive staining in both species. In addition, double staining was used to demonstrate colocalization of both ERalpha and ERbeta in efferent ductules of both species. The specificity of antibodies was demonstrated by Western blot analysis. This study reveals a differential localization of ERalpha and ERbeta in male dog and cat reproductive tracts, demonstrating more intensive expression of ERbeta than ERalpha. However, as in other species, the efferent ductules remained the region of highest concentration of ERalpha. PMID- 11906938 TI - Heat shock-induced apoptosis in preimplantation bovine embryos is a developmentally regulated phenomenon. AB - Apoptosis is a form of cell death that can function to eliminate cells damaged by environmental stress. One stress that can compromise embryonic development is elevated temperature (i.e., heat shock). For the current studies, we hypothesized that heat shock induces apoptosis in bovine embryos in a developmentally regulated manner. Studies were performed to 1) determine whether heat shock can induce apoptosis in preimplantation embryos, 2) test whether heat-induced apoptosis is developmentally regulated, 3) evaluate whether heat shock-induced changes in caspase activity parallel patterns of apoptosis, and 4) ascertain whether exposure to a mild heat shock can protect embryos from heat-induced apoptosis. As determined by TUNEL reaction, exposure of bovine embryos > or =16 cells on Day 5 after insemination to 41 or 42 degrees C for 9 h increased the percentage of cells undergoing apoptosis. In addition, there was a duration dependent increase in the proportion of blastomeres that were apoptotic when embryos were exposed to temperatures of 40 or 41 degrees C, which are more characteristic of temperatures experienced by heat-stressed cows. Heat shock also increased caspase activity in Day 5 embryos. However, heat shock did not induce apoptosis in 2- or 4-cell embryos, nor did it increase caspase activity in 2-cell embryos. The apoptotic response of 8- to 16-cell-stage bovine embryos to heat shock depended upon the day after insemination that heat shock occurred. When 8- to 16-cell embryos were collected on Day 3 after insemination, heat shock of 41 degrees C for 9 h did not induce apoptosis. In contrast, when 8- to 16-cell embryos were collected on Day 4 after insemination and exposed to heat shock, there was an increase in the percentage of cells undergoing apoptosis. Exposure of 8- to 16-cell embryos at Day 4 to a mild heat shock of 40 degrees C for 80 min blocked the apoptotic response to a subsequent, more-severe heat shock of 41 degrees C for 9 h. In conclusion, apoptosis is a developmentally acquired phenomenon that occurs in embryos exposed to elevated temperature, and it can be prevented by induced thermotolerance. PMID- 11906939 TI - Nuclear-cytoplasmic "tug of war" during cloning: effects of somatic cell nuclei on culture medium preferences of preimplantation cloned mouse embryos. AB - Cloning by somatic cell nuclear transfer is critically dependent upon early events that occur immediately after nuclear transfer, and possibly additional events that occur in the cleaving embryo. Embryo culture conditions have not been optimized for cloned embryos, and the effects of culture conditions on these early events and the successful initiation of clonal development have not been examined. To evaluate the possible effect of culture conditions on early cloned embryo development, we have compared a number of different culture media, either singly or in sequential combinations, for their ability to support preimplantation development of clones produced using cumulus cell nuclei. We find that glucose is beneficial during the 1-cell stage when CZB medium is employed. We also find that potassium simplex optimized medium (KSOM), which is optimized to support efficient early cleavage divisions in mouse embryos, does not support development during the 1-cell or 2-cell stages in the cloned embryos as well as other media. Glucose-supplemented CZB medium (CZB-G) supports initial development to the 2-cell stage very well, but does not support later cleavage stages as well as Whittten medium or KSOM. Culturing cloned embryos either entirely in Whitten medium or initially in Whittens medium and then changing to KSOM at the late 4 cell/early 8-cell stage produces consistent production of blastocysts at a greater frequency than using CZB-G medium alone. The combination of Whitten medium followed by KSOM resulted in an increased number of cells per blastocyst. Because normal embryos do not require glucose during the early cleavage stages and develop efficiently in all of the media employed, these results reveal unusual culture medium requirements that are indicative of altered physiology and metabolism in the cloned embryos. The relevance of this to understanding the kinetics and mechanisms of nuclear reprogramming and to the eventual improvement of the overall success in cloning is discussed. PMID- 11906940 TI - Evaluation of natural killer cell recruitment to embryonic attachment sites during early porcine pregnancy. AB - Specialized natural killer (NK) lymphocytes are a feature of the pregnant uterus in humans and rodents. Conceptus-mediated recruitment of uterine (u)NK cells in the pig was proposed based on evidence that elevated uNK activity was temporally associated with increased leukocyte density in endometrium underlying conceptuses. The objective of this study was to determine whether uNK cells were more abundant at embryonic attachment sites during the early postattachment period. Mononuclear leukocytes were isolated from endometrium at attachment sites versus between attachment sites, and expression of CD16, a marker for NK cells, was assessed by flow cytometry. CD16 binding was normalized to leukocyte numbers in each sample. CD16+ small lymphocytes were more frequent in uterus than in blood (41% +/- 2% versus 26% +/- 4%). Differences between pregnant and luteal phase uterus (43% +/- 2% versus 31% +/- 7%, respectively) were not statistically significant. In pregnant animals, CD16+ lymphocytes were slightly but significantly more abundant in uterus at attachment sites versus between attachment sites at Days 15-17, 21-22, and 25-28. Before normalization, CD16+ large, granular cells were more abundant at attachment sites versus between attachment sites; however, these differences were removed when data were normalized according to leukocyte numbers. Further characterization showed that the proportion of large granular leukocytes expressing CD8, reactive with NK cells and T cell subsets, was 2-fold higher in pregnant uterus than in maternal blood. These results raise the possibility that uNK cells resembling those in blood may be transformed into larger, more granulated forms in the uterine microenvironment. PMID- 11906942 TI - Vitelline envelope of Bufo arenarum: biochemical and biological characterization. AB - Vitelline envelopes (VEs) of Bufo arenarum were isolated in order to study their composition and their role in fertilization. VEs are composed of four glycoproteins, with molecular masses of 120, 75, 41, and 38 kDa. To characterize its biological properties, we quantitatively determined sperm-VE binding and the induction of the acrosome reaction. Heterologous binding of B. arenarum sperm to Xenopus laevis VE components was observed with about one-third the efficiency of homologous binding. Equivalent binding of X. laevis sperm to the B. arenarum VE was observed. When B. arenarum sperm were incubated with fluorescein isothiocyanate-labeled VE, the labeled glycoproteins bound to the anterior end of the sperm head, showing a lateral distribution. Induction of the acrosome reaction was evaluated by incubating sperm in hypotonic saline media with VE glycoproteins. VEs induced the acrosome reaction in a time- and concentration dependent manner. The acrosome reaction was maximal after 10 min. The half maximal effect was obtained at a glycoprotein concentration of 1 microg/ml. Specificity was determined using fertilization envelope glycoproteins, which failed to induce the acrosome reaction. The B. arenarum VE is biochemically similar to other egg envelopes. It also seems that its biological properties are similar to other species in regard to sperm binding and induction of the acrosome reaction. However, as far as we are aware, this is the first observation of the VE inducing the sperm acrosome reaction in amphibians. The relatively small differences observed in heterologous sperm-VE binding in X. laevis and B. arenarum are inconsistent with the current paradigm that species specificity in fertilization is regulated at the sperm-VE binding step. PMID- 11906941 TI - Analysis of the roles of RGD-binding integrins, alpha(4)/alpha(9) integrins, alpha(6) integrins, and CD9 in the interaction of the fertilin beta (ADAM2) disintegrin domain with the mouse egg membrane. AB - Fertilin beta (also known as ADAM2), a mammalian sperm protein that mediates gamete cell adhesion during fertilization, is a member of the ADAM protein family whose members have disintegrin domains with homology to integrin ligands found in snake venoms. Fertilin beta utilizes an ECD sequence within its disintegrin domain to interact with the egg plasma membrane; the Asp is especially critical. Based on what is known about different integrin subfamilies and their ligands, we sought to characterize fertilin beta binding sites on mouse eggs, focusing on integrin subfamilies that recognize short peptide sequences that include an Asp residue: the alpha(5)/alpha(8)/alpha(v)/alpha(IIb) or RGD-binding subfamily (alpha(5)beta(1), alpha(8)beta(1), alpha(V)beta(1), alpha(V)beta(3), alpha(V)beta(5), alpha(V)beta(6), alpha(V)beta(8), and alpha(IIb)beta(3)) and the alpha(4)/alpha(9) subfamily (alpha(4)beta(1), alpha(9)beta(1), and alpha(4)beta(7)). We tested peptide sequences known to perturb interactions mediated by these integrins in two different assays for fertilin beta binding. Peptides with the sequence MLDG, which perturb alpha(4)/alpha(9) integrin mediated interactions, significantly inhibit fertilin beta binding to eggs, which suggests a role for a member of this integrin subfamily as a fertilin beta receptor. RGD peptides, which perturb alpha(5)/alpha(8)/alpha(v)/alpha(IIb) integrin-mediated interactions, have partial inhibitory activity. The anti alpha(6) antibody GoH3 has little or no inhibitory activity. An antibody to the integrin-associated tetraspanin protein CD9 inhibits the binding of a multivalent presentation of fertilin beta (immobilized on beads) but not soluble fertilin beta, which we speculate has implications for the role of CD9 in the strengthening of fertilin beta-mediated cell adhesion but not in initial ligand binding. PMID- 11906943 TI - Characterization of a prolactin-regulated gene in reproductive tissues using the prolactin receptor knockout mouse model. AB - Prolactin (PRL) exerts pleiotropic physiological effects in various cells and tissues, although it is mainly considered as a regulator of reproduction and cell growth. Null mutation of the prolactin receptor (PRLR) gene leads to female sterility due to a failure of embryo implantation. Using this mouse model and the method of mRNA differential display, we identified PRL target genes that are regulated during the peri-implantation period. We characterized 1 among the 45 isolated genes, UA-3, which is regulated in the uterus as well as in the ovary during early pregnancy. This gene corresponds to a P311 mouse cDNA that was originally identified for its high expression in late-stage embryonic brain and adult cerebellum. We report here that UA-3 is present in numerous tissues as well as in ovary and uterus at the site of blastocyst apposition, and that its expression is hormonally regulated. Moreover, in situ hybridization reveals high expression in ovarian granulosa cells and in uterine epithelium. Recently, it has been suggested that P311 expression is tightly regulated at several levels by mechanisms that control cellular growth, transformation, motility, or a combination of these. Taken together, these results suggest that P311 could be involved in these processes during pregnancy, although its function remains to be clearly established. PMID- 11906944 TI - Antagonism of P2X3-containing channels: commentary on Spelta et al. PMID- 11906945 TI - Cyclo-oxygenase-2 inhibition and exacerbation of myocardial dysfunction- protection with nitric oxide? PMID- 11906946 TI - Endothelial dysfunction in aging animals: the role of poly(ADP-ribose) polymerase activation. AB - Recent work has demonstrated the production of reactive oxygen and nitrogen species in the vasculature of aging animals. Oxidant induced cell injury triggers the activation of nuclear enzyme poly(ADP ribose) polymerase (PARP) leading to endothelial dysfunction in various pathophysiological conditions (reperfusion, shock, diabetes). Here we studied whether the loss of endothelial function in aging rats is dependent upon the PARP pathway within the vasculature. Young (3 months-old) and aging (22 months-old) Wistar rats were treated for 2 months with vehicle or the PARP inhibitor PJ34. In the vehicle-treated aging animals there was a significant loss of endothelial function, as measured by the relaxant responsiveness of vascular rings to acetylcholine. Treatment with PJ34, a potent PARP inhibitor, restored normal endothelial function. There was no impairment of the contractile function and endothelium-independent vasodilatation in aging rats. Furthermore, we found no deterioration in the myocardial contractile function in aging animals. Thus, intraendothelial PARP activation may contribute to endothelial dysfunction associated with aging. PMID- 11906947 TI - Tachykinin NK1 and NK2 receptors mediate inhibitory vs excitatory motor responses in human isolated corpus cavernosum and spongiosum. AB - Motor effects produced by tachykinins were studied in human isolated corpus spongiosum and cavernosum. In quiescent preparations neurokinin A caused potent contractions (pD(2)=8.3 - 7.9 respectively) prevented by the NK(2) receptor selective antagonist nepadutant, whereas [Sar(9)]SP sulfone and senktide (NK(1) and NK(3) receptor-selective agonists) produced no effect or spare contractions. In KCl-precontracted corpus spongiosum septide (pD(2)=7.1) and [Sar(9)]SP sulfone (pD(2)=7.7) produced tetrodotoxin-resistant relaxations, abolished by the tachykinin NK(1) receptor-selective antagonist SR 140333. [Sar(9)]SP sulfone (1 microM) produced similar relaxations in precontracted corpus cavernosum. Electrical field stimulation (EFS) elicited tetrodotoxin-sensitive relaxations, which were additive to those produced by [Sar(9)]SP sulfone. N(omega)-nitro-L arginine (L-NOARG) totally prevented both [Sar(9)]SP sulfone- and EFS-induced relaxations. These results show that tachykinin NK(1) and NK(2) receptors mediate opposite motor effects in human penile tissues, suggesting a possible modulatory role of tachykinins on smooth muscle tone in these organs. PMID- 11906948 TI - Increased function of inhibitory neuronal M2 muscarinic receptors in trachea and ileum of diabetic rats. AB - 1. Release of acetylcholine from parasympathetic nerves is inhibited by neuronal M(2) muscarinic receptors. The effects of streptozotocin-induced diabetes on prejunctional M(2) and postjunctional M(3) muscarinic receptor function in rat trachea and ileum were investigated in vitro. 2. Neuronal M(2) muscarinic receptor function was tested by measuring the ability of an agonist, pilocarpine, to inhibit and an antagonist, methoctramine, to potentiate electrical field stimulation (EFS)-induced contraction of trachea and ileum. Concentration response curves to pilocarpine and methoctramine were shifted to the left in both to a greater degree in diabetics than controls. 3. In trachea, post-junctional M(3) muscarinic receptor function was increased since maximum contractile responses to the muscarinic agonists acetylcholine and carbachol were greater in diabetics than controls. This increase offset the increased function of the inhibitory neuronal M(2) muscarinic receptors since EFS-induced, frequency dependent contraction was equal in control and diabetic rats. 4. In contrast, post-junctional M(3) muscarinic receptor function was unchanged by diabetes since concentration-response curves to acetylcholine and carbachol were not different between groups. Thus, EFS-induced contractions of the ileum were decreased in diabetics versus controls. 5. In conclusion, inhibitory M(2) muscarinic receptors on parasympathetic nerves in the trachea and ileum are hyperfunctional in diabetic rats. The function of post-junctional M(3) muscarinic receptors in the trachea, but not the ileum, is also increased in diabetes. 6. The dysfunction of inhibitory, neuronal M(2) muscarinic receptors in the airways may protect against hyperreactivity and in the ileum may contribute to gastrointestinal dysmotility associated with diabetes. PMID- 11906949 TI - Modulators of internal Ca2+ stores and the spontaneous electrical and contractile activity of the guinea-pig renal pelvis. AB - 1. The role of internal Ca(2+) stores in the generation of the rhythmic electrical and contractile activity in the guinea-pig proximal renal pelvis was examined using intracellular microelectrode and muscle tension recording techniques. 2. Ryanodine (30 microM) transiently increased contraction amplitude, while caffeine (0.5 - 3 mM) reduced contraction amplitude and frequency. Contractility was also reduced by 2-aminoethoxy-diphenylborate (2-APB 60 microM), xestospongin C (1 microM), U73122 (5 microM) and neomycin (4 mM), blockers of IP(3)-dependent release from Ca(2+) stores. 3. 60 mM K(+) saline-evoked contractions were reduced by caffeine (1 mM), U73122 (5 microM) and neomycin (4 mM), but little affected by ryanodine or 2-APB (60 microM). 4. Spontaneous action potentials consisting of an initial spike followed by a long plateau were recorded (frequency 8.6+/-1.0 min(-1)) in small urothelium-denuded strips of proximal renal pelvis. 5. Action potential discharge was blocked in 75 and 35% of cells by 2-APB (60 microM) and caffeine (1 mM), respectively. In the remaining cells, only a truncation of the plateau phase was observed. 6. Cyclopiazonic acid (CPA 10 microM for 10 - 180 min), blocker of CaATPase, transiently increased contraction frequency and amplitude. Action potential durations were increased 3.6 fold. Contraction amplitude and frequency slowly declined during a prolonged (>60 min) CPA exposure. 7. We conclude that the action potential in caffeine sensitive cells and the shoulder component of caffeine-insensitive action potential arise from the entry of Ca(2+) through Ca(2+) channels. The inhibitory actions of modulators of internal Ca(2+) release were partially explained by a blockade of Ca(2+) entry. PMID- 11906950 TI - Characterization of the human fMLP receptor in neutrophils and in Xenopus oocytes. AB - 1. N-formyl peptides (e.g. fMLP; N-formyl-L-methionyl-L-leucyl-phenylalanine) are potent mediators for inflammatory reactions. We report functional expression in Xenopus oocytes of human fMLP-R98 cDNA, without co-expression of the promiscuous G-protein subunit, Galpha-16. 2. Stimulation of voltage-clamped oocytes (-70 mV) with fMLP produced a dose-dependent biphasic inward current with fast and slow components. Analysis using GTP-gamma-S and cholera and pertussis toxins suggested these currents are mediated by an endogenous G-protein of the Gq family. 3. The fast current reversed at -25 mV and was blocked by SITS (4-acetamido-4' isothiocyanatostilbene-2,2'-disulphonic acid), suggesting the current is carried by Cl(-). The slow current showed weak inward rectification, was Ca(2+)-dependent and blocked by Cd(2+), 4-AP (4-aminopyridine) and haloperidol, suggesting activation of a mixed population of cation channels. 4. Comparative experiments with human neutrophils using flow cytometric analysis showed that the proportion of neutrophils activated by fMLP was reduced in the presence of SITS, in the absence of external calcium and in the presence of Cd(2+), TEA (tetraethylammonium) and haloperidol but not 4-AP. In addition, the oxidative burst from activated neutrophils was reduced by SITS and by the absence of external calcium but not by Cd(2+), TEA, 4-AP or haloperidol. 5. We suggest that in human neutrophils activation by fMLP is dependent on store-operated calcium influx that appears to be regulated by Cl(-) channels and linked, in part, to non selective cation channels. PMID- 11906951 TI - Store depletion-induced calcium influx in rat cerebellar astrocytes. AB - 1. In rat cerebellar astrocytes, intracellular Ca(2+) store depletion by receptor agonists or sarco(endo)plasmic reticulum Ca(2+) ATPase inhibitors induced a transient increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)) in the absence of extracellular Ca(2+) and a sustained increase in its presence. 2. After 10 min treatment with thapsigargin, the [Ca(2+)](i) was unaffected by removal of thapsigargin, but fell rapidly to the basal level when extracellular Ca(2+) was removed, suggesting the involvement of capacitative Ca(2+) entry (CCE); this effect was not seen until cells had been exposed to thapsigargin for at least 2 min. 3. Using the whole cell voltage clamp technique, a 60-100 pA inward current was activated by store depletion, the reversal potential ranging from -5 to 0 mV. 4. When extracellular Na(+) was isotonically replaced by Tris, the thapsigargin-induced [Ca(2+)](i) increase was enhanced, while the inward current was reduced, indicating that store-operated Ca(2+) channels were permeable to Na(+); however, they were not permeable to Sr(2+) or Ba(2+). 5. Thapsigargin-induced CCE remained the same in the presence of nifedipine, La(3+) or Cd(2+), while it was inhibited in the presence of SK&F96365. 6. In cerebellar astrocytes, inhibition of protein serine/threonine phosphorylation promoted CCE. 7. In conclusion, in rat cerebellar astrocytes, store depletion activated a CCE via channels which were permeable to Ca(2+) and Na(+) and regulated by phosphorylation. PMID- 11906952 TI - Desferrithiocin is a more potent antineoplastic agent than desferrioxamine. AB - Desferrithiocin (DFT) is an orally effective Fe chelator, with a similar high affinity and selectivity for Fe to desferrioxamine (DFO), which has been shown clinically to possess antineoplastic activity. In this study, DFT was assessed for antineoplastic potential in hepatocellular carcinoma cell lines (HCC). This was done as there are few treatments for this aggressive neoplasm. The effects of DFT on cell proliferation, cell cycle progression, Fe uptake and toxicity were examined. To establish whether DFT was selective for cancer cells a comparison was made with normal (non-proliferating) hepatocytes and non-tumorigenic (proliferating) fibroblasts (SWISS-3T3). DFT was a potent inhibitor of HCC proliferation (IC(50) approximately 40 microM). DFO also inhibited HCC proliferation under the same conditions, but was much less active (IC(50)=110 - 210 microM). When saturated with Fe, the activity of DFT, like DFO, was greatly diminished, suggesting it may act by depriving the cells of Fe or inactivating essential Fe pool(s). Indeed DFT rapidly decreased Fe uptake from Tf-(59)Fe by hepatoma cells and also by normal hepatocytes. However, DFT (and DFO) had much less effect on cell survival in hepatocytes and fibroblasts than in hepatoma cells. DFT may, like DFO, inhibit the cell cycle in the S phase of DNA synthesis. Both chelators showed low toxicity. These results indicate that DFT has potent antineoplastic activity in HCC. Further investigation into the DFT class of Fe chelators seems warranted, particularly in view of its high activity in relation to DFO, a chelator which is already in clinical trial for neuroblastoma. PMID- 11906953 TI - An electrophysiological study of muscarinic and nicotinic receptors of rat paratracheal ganglion neurons and their inhibition by Z-338. AB - 1. To study the mechanisms involved in the action of Z-338, a newly synthesized gastroprokinetic agent, experiments were performed with the paratracheal ganglion cells acutely dissociated from 2-week-old Wistar rats. The effects of Z-338 on both nicotinic and muscarinic responses of the ganglion cells were studied by nystatin perforated patch recording configuration under the current- and voltage clamp conditions. 2. Acetylcholine (ACh) or nicotine, and muscarine or oxotremorine-M (OX-M) induced membrane depolarization with rapid and slow time courses respectively, followed by repetitive generation of action potentials in the ganglion cell. Corresponding to the membrane depolarization induced by cholinergic agents, ACh induced biphasic inward currents with rapid and slow time courses under the voltage-clamp condition. Nicotine and muscarine or OX-M evoked inward currents with rapid and slow time courses, respectively. The rapid and slow inward currents were accompanied by increase and decrease in the membrane conductance, respectively. In addition, OX-M dose-dependently suppressed the M type K(+) current evoked in response to hyperpolarizing voltage-steps from V(H) of -25 mV to -50 mV, indicating that the activation of muscarinic acetylcholine receptors inhibits M-type K(+) current, thus inducing inward current in the ganglion cell. 3. Z-338 competitively suppressed the inward currents induced by OX-M through M(1) ACh receptor, and uncompetitively suppressed the currents induced by nicotine. 4. The inhibitory actions of Z-338 on the membrane depolarization and corresponding inward currents mediated by M(1)-muscarinic and neuronal nicotinic ACh receptors in the isolated ganglion cells were discussed in relation to the inhibitory actions on autoreceptors in the parasympathetic nerve terminals, which would explain the gastroprokinetic actions of Z-338. PMID- 11906954 TI - Postnatal changes in beta-adrenoceptors in the lung and the effect of hypoxia induced pulmonary hypertension of the newborn. AB - 1. beta-adrenoceptor activation leads to pulmonary vasodilatation. The increase in circulating catecholamines at birth may assist the postnatal fall in vascular resistance by their activation. To study beta(1)- and beta(2)-adrenoceptors during postnatal adaptation, we used [(125)I]-iodocyanopindolol (ICYP) binding to lung membranes and sections to quantify and locate the binding sites in piglets from birth to 14 days of age and compared them with those in adult pigs. In addition, pulmonary hypertension was induced in newborn piglets by hypobaric hypoxia. 2. In lung membranes the equilibrium dissociation constant (K(d)) did not change with age for total beta-adrenoceptors or for beta(2)-adrenoceptors, but there was a significant increase in maximum binding sites (B(max)) between birth and 3 days of age. On tissue sections, B(max) increased between 3 days and adulthood with no change in K(d). 3. Binding sites of beta(1)- and beta(2) adrenoceptors were localized to the bronchial epithelium, to endothelium of extra and intra-pulmonary arteries and to lung parenchyma. Total beta-adrenoceptor density increased with age at all locations (P<0.05 - 0.01). At birth intrapulmonary arteries showed no binding, beta(2)-adrenoceptors appeared on day 1 and increased up to 14 days of age. beta(1)-adrenoceptors appeared by 3 days of age and increased with age. 4. Hypobaric hypoxia from birth led to attenuation in the normal postnatal increase in receptor number, but hypoxia from 3 - 6 days did not decrease receptor density. 5. The normal postnatal increase in beta adrenoceptors suggests a potential for catecholamine induced dilatation in the lung during adaptation which is attenuated in pulmonary hypertension. PMID- 11906955 TI - A comparison of the efficacy of carbamazepine and the novel anti-epileptic drug levetiracetam in the tetanus toxin model of focal complex partial epilepsy. AB - 1. The tetanus toxin seizure model, which is associated with spontaneous and intermittent generalized and non-generalized seizures, is considered to reflect human complex partial epilepsy. The purpose of the present study was to investigate and compare the anticonvulsant effects of carbamazepine with that of levetiracetam, a new anti-epileptic drug in this model. 2. One microl of tetanus toxin solution (containing 12 mLD(50) microl(-1) of tetanus toxin) was placed stereotactically into the rat left hippocampus resulting in generalized and non generalized seizures. 3. Carbamazepine (4 mg kg(-1) h(-1)) and levetiracetam (8 and 16 mg kg(-1) h(-1)) were administered during a 7 day period via an osmotic minipump which was placed in the peritoneal cavity. Carbamazepine (4 mg kg(-1) h( 1)) exhibited no significant anticonvulsant effect, compared to control, when the entire 7 day study period was evaluated but the reduction in generalized seizures was greater (35.5%) than that for non-generalized seizures (12.6%). However, during the first 2 days of carbamazepine administration a significant reduction in both generalized seizure frequency (90%) and duration (25%) was observed. Non generalized seizures were unaffected. This time-dependent anticonvulsant effect exactly paralleled the central (CSF) and peripheral (serum) kinetics of carbamazepine in that steady-state concentrations declined over time, with the highest concentrations achieved during the first 2 days. Also there was a significant 27.3% reduction in duration of generalized seizures during the 7 day study period (P=0.0001). 4. Levetiracetam administration (8 and 16 mg kg(-1) h( 1)) was associated with a dose-dependent reduction in the frequency of both generalized (39 v 57%) and non-generalized (36 v 41%) seizures. However, seizure suppression was more substantial for generalized seizures. Also a significant dose-dependent reduction in overall generalized seizure duration was observed. 5. These data provide experimental evidence for the clinical efficacy of levetiracetam for the management of patients with complex partial seizures. Furthermore, levetiracetam probably does not act by preventing ictogenesis per se but acts to reduce seizure severity and seizure generalization. PMID- 11906956 TI - Enhanced neuronal damage by co-administration of quinolinic acid and free radicals, and protection by adenosine A2A receptor antagonists. AB - 1. Quinolinic acid may be an important endogenous excitotoxin, but its concentrations in brain are low. We have therefore attempted to determine whether its neurotoxicity can be increased by the simultaneous presence of free radicals. 2. Quinolinic acid was injected into the hippocampus of anaesthetized rats at doses of 40 and 80 nmols which produced little neuronal loss, and 120 nmols which produced over 90% neuronal loss. 3. A mixture of xanthine and xanthine oxidase, a known source of free radical reactive oxygen species, also generated little damage alone, but killed over 80% of CA1 neurons when combined with 80 nmols of quinolinic acid. Similarly, the nitric oxide donor S-nitroso-N acetylpenicillamine (SNAP) potentiated the damage produced by quinolinic acid. 4. The glutamate antagonist 5,7-dichlorokynurenic acid prevented the damage produced by 120 nmols of quinolinic acid, but not that produced by quinolinic acid plus xanthine/xanthine oxidase, indicating that damage was not simply the result of free radical enhancement of NMDA receptor activation. 5. Three chemically dissimilar antagonists at adenosine A(2A) receptors prevented the damage caused by quinolinic acid and xanthine/xanthine oxidase or by quinolinic acid plus SNAP. 6. It is concluded that reactive oxygen species can potentiate the neurotoxicity of quinolinic acid. The site of interaction is probably distal to the NMDA receptor. Blockade of adenosine A(2A) receptors can protect against this combined damage, suggesting potential value in the prevention of brain damage. PMID- 11906957 TI - Anti-secretory properties of non-peptide somatostatin receptor agonists in isolated rat colon: luminal activity and possible interaction with P glycoprotein. AB - 1. The diverse physiological actions of somatostatin are mediated by a family of G-protein coupled receptors (SSTRs). Several peptide analogues of somatostatin such as octreotide have been developed for therapeutic use, including treatment of gastrointestinal disorders such as secretory diarrhoea. However, their development as anti-diarrhoeal agents has been limited by poor oral bioavailability, necessitating parenteral administration. This in vitro study investigated the anti-secretory potential of a group of novel, non-peptide, somatostatin-receptor agonists that selectively activate specific SSTR subtypes to assess their potential for oral administration. 2. The ability of the agonists to inhibit forskolin-stimulated chloride secretion was measured using a sensitive bioassay system in isolated rat colonic mucosa. 3. The SSTR-2 selective agonist, L-779,976 was 10-times more potent than octreotide as an inhibitor of secretion when added to the basolateral surface of rat colon. Non-peptide agonists selective for SSTR1 (L-797,591), SSTR3 (L-796,778), SSTR4 (L-803,087) or SSTR5 (L 817,818) showed little or no anti-secretory activity in this preparation. 4. L 779,976 was able to inhibit secretion when applied to the luminal surface at sub micromolar concentrations suggesting that it can cross the colonic epithelium. The anti-secretory potency of luminal L-779,976 was increased 3 fold in the presence of GF120918, a known inhibitor of P-glycoprotein. 5. Non-peptide somatostatin receptor agonists may provide a basis for the development of new, orally available anti-diarrhoeal therapies. PMID- 11906958 TI - Differential sensitivity to tetrodotoxin and lack of effect of prostaglandin E2 on the pharmacology and physiology of propagated action potentials. AB - 1. We have studied the effects of prostaglandin E(2) (PGE(2)) on action potential propagation in the isolated, desheathed vagus and saphenous nerves of rats using an extracellular grease gap recording method. 2. PGE(2) evoked a small depolarization of vagus nerves but had no effect on the stimulation threshold, size or latency of either the A wave (corresponding to conduction in A fibres) or the C wave (corresponding to conduction in C fibres) of the compound action potential (CAP) recorded from either vagus or saphenous nerves. 3. Lidocaine (0.01 - 10 mM) reduced all components of the CAP of both vagus and saphenous nerves. PGE(2) had no significant effect on the sensitivity of any component of the CAP to lidocaine. 4. Tetrodotoxin (TTX, 10 microM) blocked completely both the A wave and the C wave of the CAP in either vagus or saphenous nerves. 5. In saphenous nerve preparations the A wave was blocked by lower concentrations of TTX than the C wave or any component of the CAP in vagus nerve preparations which suggests that somatosensory A fibres express a different sub-type of TTX sensitive voltage-gated sodium channel (VGSC) than somatosensory C-fibres or visceral sensory fibres. 6. Chemical activation of VGSCs with veratridine (10 or 50 microM) induced a depolarization in either nerve. The depolarization induced by 50 microM veratridine was blocked by 10 microM TTX. 7. Although TTX insensitive VGSCs are expressed by some vagal and some somatosensory neurones they do not appear to be expressed functionally in the axons. PMID- 11906959 TI - Tonic activity of the rat adipocyte A1-adenosine receptor. AB - 1. Adipocyte A(1)-adenosine receptors (A(1) AdoR) tonically inhibit adenylyl cyclase and lipolysis. Three potential explanations for tonic activity of A(1)AdoR of rat epididymal adipocytes were investigated: high affinity of adenosine for the receptor, efficient coupling of receptor activation to response, and spontaneous activity of the receptor in the absence of agonist. 2. The affinity of adenosine for the adipocyte A(1)AdoR was determined as 4.6 microM by analysis of effects of an irreversible receptor antagonist on agonist concentration-response relationships. In contrast, the potency of adenosine to decrease cyclic AMP in isolated adipocytes was 1.4 nM. 3. Occupancy by agonist of the A(1)AdoR was efficiently coupled to functional response (decrease of adipocyte cyclic AMP content). Activation by adenosine of less than 1% of A(1)AdoRs caused a near-maximal decrease of cyclic AMP in adipocytes. Thus the receptor reserve for adenosine to decrease cyclic AMP content of adipocytes was greater than 99%. 4. Affinities and receptor reserves for other A(1)AdoR agonists were determined. Agonists appeared to differ more in their affinity for the receptor than in their intrinsic efficacy to activate it. 5. A(1)AdoRs were inactive in the absence of agonist. 6. It is concluded that adipocyte A(1)AdoR are tonically activated by endogenous adenosine at nanomolar concentrations. The expression of a high density of A(1)AdoR that are efficiently coupled to a functional response enables the adipocyte to respond with high sensitivity to the low-affinity agonist, adenosine. Adipocytes may be a model for cells whose functions are tonically modulated by adenosine present in the interstitium of well-oxygenated tissues. PMID- 11906961 TI - 2-Chloroadenosine but not adenosine induces apoptosis in rheumatoid fibroblasts independently of cell surface adenosine receptor signalling. AB - 1. The apoptotic effect of adenosine and its analogues was studied in fibroblast like synoviocytes derived from rheumatoid arthritis patients (RA-FLSs). Evoked cell death was quantitatively examined by assessing DNA fragmentation using an enzyme-liked immunosorbent assay and by measuring phosphatidylserine exposure through flow cytometric analysis of annexin V binding. 2. Exposing cells for 24 h to 2-chloroadenosine (2-CADO), a nonspecific, adenosine deaminase (ADA) resistant, adenosine receptor (AdoR) agonist, induced DNA fragmentation, and thus apoptosis, in RA-FLSs at concentrations > or =50 microM. By contrast, incubation with adenosine for up to 72 h did not evoke DNA fragmentation, even in the presence of ADA inhibitor coformycin and nucleoside transporter inhibitor nitrobenzylmercaptopurin (NBMPR). Transcription of all four AdoR isoforms was detected in RA-FLSs; nevertheless selective AdoR agonists similarly failed to induce DNA fragmentation. 3. DNA fragmentation evoked by 2-CADO was inhibited by NBMPR and by 5'-iodotubercidin, an adenosine kinase inhibitor, but not by xanthine amine congener, an A(1) and A(2) receptor antagonist, or by selective AdoR antagonists. 4. The nonspecific caspase inhibitor benzyloxycarbonyl-Val-Ala Asp fluoromethyl ketone abolished the apoptotic effect of 2-CADO. 5. These results suggest that 2-CADO induces apoptosis in RA-FLSs independently of AdoR mediated signalling. Instead, 2-CADO, but not adenosine, is taken up into RA-FLSs via human equilibrative nucleoside transporter-1, where it is phosphorylated by adenosine kinase. The resultant phospho-2-CADO induces DNA fragmentation by activating a caspase pathway. PMID- 11906960 TI - Urocortin-induced endothelium-dependent relaxation of rat coronary artery: role of nitric oxide and K+ channels. AB - 1. The mechanisms underlying the vasodilator response to urocortin are incompletely understood. The present study was designed to examine the role of endothelial nitric oxide and Ba(2+)-sensitive K(+) channels in the endothelium dependent component of urocortin-induced relaxation in the rat left anterior descending coronary artery. 2. Urocortin induced both endothelium-dependent and independent relaxation with respective pD(2) of 8.64+/-0.03 and 7.90+/-0.10. Removal of endothelium reduced the relaxing potency of urocortin. In rings pretreated with 10(-4) M N(G)-nitro-L-arginine methyl ester, 10(-5) M methylene blue or 10(-5) M ODQ, the urocortin-induced relaxation was similar to that observed in endothelium-denuded rings. L-Arginine (5x10(-4) M) antagonized the effect of N(G)-nitro-L-arginine methyl ester. 3. The relaxant response to urocortin was reduced in endothelium-intact rings preconstricted by 3.5x10(-2) M K(+) and abolished when extracellular K(+) was raised to 5x10(-2) M. Pretreatment with 10(-4) M BaCl(2) significantly inhibited urocortin-induced relaxation. Combined treatment with 10(-4) M BaCl(2) plus 10(-4) M N(G)-nitro-L-arginine methyl ester did not cause further inhibition. In urocortin (10(-8) M)-relaxed rings, BaCl(2) induced concentration-dependent reversal in vessel tone. Tertiapin Q (10(-6) M) also attenuated urocortin-induced relaxation. In contrast, BaCl(2) did not alter urocortin-induced relaxation in endothelium-denuded rings. 4. In endothelium-denuded rings, hydroxylamine- and nitroprusside-induced relaxation was inhibited by 10(-4) M BaCl(2), but not by 10(-6) M tertiapin-Q. 5. The endothelium of the coronary artery was moderately stained with the antiserum against urocortin. 6. Taken together, the present results indicate that the urocortin-induced endothelium-dependent relaxation of rat coronary arteries is likely attributable to endothelial nitric oxide and subsequent activation of Ba(2+)- or tertiapin-Q-sensitive K(+) channels. The urocortin-induced endothelium dependent relaxation appears to be mediated by cyclic GMP-dependent mechanisms. PMID- 11906963 TI - Effects of FPL 64176 on Ca transients in voltage-clamped rat ventricular myocytes. AB - 1. The L-type Ca channel agonist FPL 64176 increased the amplitude of both Ca currents and Ca transients elicited from isolated voltage clamped rat ventricular myocytes far more than it increased the rate of rise of the Ca transients. Consequently, the gain function relating the amplitude of peak Ca current to Ca transient rate of rise was greatly reduced at all potentials. 2. Furthermore, an increase in this gain function normally observed at negative potentials is abolished by FPL 64716. 3. Despite slowing the rate of decline of Ca transients, FPL 64176, at the concentration of 1 microM used throughout, had no direct effect on sarcoplasmic reticulum (SR) Ca uptake or release using isolated cardiac membranes. 4. Arguments based on results presented here and elsewhere suggest that decreased gain was not due to increased ryanodine receptor adaptation or inactivation, to decreased L-type single channel current, to decreased SR Ca content, or to decreased synchronization of release. Decreased gain instead appears to reflect a form of decrease in coupling efficiency due either to differential effects of long openings on whole cell currents as opposed to the Ca release the long openings trigger or to some compensatory mechanism activated by the increased Ca trigger or resting free [Ca(2+)](i). 5. Abolition by FPL 64176 of the increased gain normally seen at negative potentials rendered it impossible to confirm or refute the claim that a single Ca ion suffices to activate ryanodine receptors. PMID- 11906962 TI - Sensitizing effects of lafutidine on CGRP-containing afferent nerves in the rat stomach. AB - 1. Capsaicin sensitive afferent nerves play an important role in gastric mucosal defensive mechanisms. Capsaicin stimulates afferent nerves and enhances the release of calcitonin gene-related peptide (CGRP), which seems to be the predominant neurotransmitter of spinal afferents in the rat stomach, exerting many pharmacological effects by a direct mechanism or indirectly through second messengers such as nitric oxide (NO). 2. Lafutidine is a new type of anti-ulcer drug, possessing both an antisecretory effect, exerted via histamine H(2) receptor blockade, and gastroprotective activities. Studies with certain antagonists or chemical deafferentation techniques suggest the gastroprotective actions of lafutidine to be mediated by capsaicin sensitive afferent nerves, but this is an assumption based on indirect techniques. In order to explain the direct relation of lafutidine to afferent nerves, we conducted the following studies. 3. We determined CGRP and NO release from rat stomach and specific [(3)H]-resiniferatoxin (RTX) binding to gastric vanilloid receptor subtype 1 (VR1), which binds capsaicin, using EIA, a microdialysis system and a radioreceptor assay, respectively. 4. Lafutidine enhanced both CGRP and NO release from the rat stomach induced by a submaximal dose of capsaicin, but had no effect on specific [(3)H]-RTX and capsaicin binding to VR1. 5. In conclusion, our findings demonstrate that lafutidine modulates the activity of capsaicin sensitive afferent nerves in the rat stomach, which may be a key mechanism involved in its gastroprotective action. PMID- 11906964 TI - Neuropeptide Y, Y1, Y2 and Y4 receptors mediate Y agonist responses in isolated human colon mucosa. AB - 1. The aim of this study was to provide a pharmacological characterization of the Y receptor types responsible for neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP) effects upon electrogenic ion transport in isolated human colonic mucosa. 2. Preparations of descending colon were voltage-clamped at 0 mV in Ussing chambers and changes in short-circuit current (I(sc)) continuously recorded. Basolateral PYY, NPY, human PP (hPP), PYY(3 - 36), [Leu(31), Pro(34)]PYY (Pro(34)PYY) and [Leu(31), Pro(34)]-NPY (Pro(34)NPY) all reduced basal I(sc) in untreated colon. Of all the Y agonists tested PYY(3 - 36) responses were most sensitive to tetrodotoxin (TTX) pretreatment, indicating that Y(2)-receptors are located on intrinsic neurones as well as epithelia in this tissue. 3. The EC(50) values for Pro(34)PYY, PYY(3 - 36) and hPP were 9.7 nM (4.0 - 23.5), 11.4 nM (7.6 - 17.0) and 14.5 nM (10.2 - 20.5) and response curves exhibited similar efficacies. The novel Y(5) agonist [Ala(31), Aib(32)]-NPY had no effect at 100 nM. 4. Y(1) receptor antagonists, BIBP3226 and BIBO3304 both increased basal I(sc) levels per se and inhibited subsequent PYY and Pro(34)PYY but not hPP or PYY(3 - 36) responses. The Y(2) antagonist, BIIE0246 also raised basal I(sc) levels and attenuated subsequent PYY(3 - 36) but not Pro(34)PYY or hPP responses. 5. We conclude that Y(1) and Y(2) receptor-mediated inhibitory tone exists in human colon mucosa. PYY and NPY exert their effects via both Y(1) and Y(2) receptors, but the insensitivity of hPP responses to either Y(1) or Y(2) antagonism, or to TTX, indicates that Y(4) receptors are involved and that they are predominantly post-junctional in human colon. PMID- 11906965 TI - Anthrapyridones, a novel group of antitumour non-cross resistant anthraquinone analogues. Synthesis and molecular basis of the cytotoxic activity towards K562/DOX cells. AB - 1. Multidrug resistance (MDR) to antitumour agents, structurally dissimilar and having different intracellular targets, is the major problem in cancer therapy. MDR phenomenon is associated with the presence of membrane proteins which belong to the ATP-binding cassette family transporters responsible for the active drug efflux leading to the decreased intracellular accumulation. 2. The search of new compounds able to overcome MDR is of prime importance. 3. Recently we have synthesized a new family of anthrapyridone compounds. The series contained derivatives modified with appropriate hydrophobic or hydrophylic substituents at the side chain. 4. The interaction of these derivatives with erythroleukemia K562 sensitive and K562/DOX resistant (overexpressing P-glycoprotein) cell lines has been examined. The study was performed using a spectrofluorometric method which allows to continuously follow the uptake and efflux of fluorescent molecules by living cells. 5. It was demonstrated that the increase in the lipophilicity of anthrapyridones favoured the very fast cellular uptake exceeding the rate of P-gp dependent efflux out of the cell. For these derivatives, very high accumulation (the same for sensitive and resistant cells) was observed and the in vitro biological data confirmed that these compounds exhibited comparable cytotoxic activity towards sensitive and P-gp resistant cell line. In contrast, anthrapyridones modified with hydrophylic substituents exhibited relatively low kinetics of cellular uptake. 6. For these derivatives decreased accumulation in resistant cells was observed and the in vitro biological data demonstrated that they were much less active against P-gp resistant cells in comparison to sensitive cells. 7. We also studied, using confocal microscopy, the intracellular distribution of anthrapyridones in NIH-3T3 cells. Our data showed that these compounds were strongly accumulated in the nucleus and lysosomes. PMID- 11906966 TI - Kinetics of antagonist actions at rat P2X2/3 heteromeric receptors. AB - 1. Currents through heteromeric P2X(2/3) receptors were evoked by applying alpha,beta-methylene-ATP to human embryonic kidney cells transfected with cDNAs encoding the P2X(2) and P2X(3) subunits. The concentration of alpha,beta methylene-ATP were < or =30 microM because higher concentrations can activate homomeric P2X(2) receptors. The kinetics of action of three structurally unrelated antagonists were studied; these were 2', 3'-O-(2,4,6,trinitrophenyl) ATP (TNP-ATP), pyridoxal-5-phosphate-6-azophenyl-2',4'-disulphonate (PPADS) and suramin. The association and dissociation rate constants were determined by pre applying the antagonist for various periods prior to the co-application of agonist and antagonist, or by changing the solution from one containing only the agonist to one containing both agonist and antagonist. The high affinity of TNP ATP for the P2X(2/3) receptor (K(D) approximately 2 nM) results from fast binding (k(+1) approximately 100 microM(-1) s(-1)) rather than slow unbinding (k(-1) approximately 0.3 s(-1)). For suramin (K(D) approximately 1 microM) the association rate constant ( approximately 1 microM(-1) s(-1)) was 100 times slower than that of TNP-ATP but the dissociation rate constant was similar (k(-1) approximately 1 s(-1)). PPADS (K(D) approximately 0.1 microM) associated and dissociated some 100 - 10,000 times more slowly than the other antagonists. PMID- 11906967 TI - The atypical 5-HT2 receptor mediating tachycardia in pithed rats: pharmacological correlation with the 5-HT2A receptor subtype. AB - 1. In pithed rats, 5-HT mediates tachycardia both directly (by 5-HT(2) receptors) and indirectly (by a tyramine-like effect). The receptor mediating tachycardia directly has been classified as an 'atypical' 5-HT(2) receptor since it was 'weakly' blocked by ketanserin. Moreover, 1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane (DOI), a 5-HT(2) agonist, failed to mimic 5-HT-induced tachycardia. Since 5-HT(2) receptors consist of 5-HT(2A), 5-HT(2B) and 5-HT(2C) subtypes, this study investigated if these subtypes mediate the above response. 2. In pithed rats, intraperitoneally (i.p.) pre-treated with reserpine (5 mg kg(-1)), intravenous (i.v.) administration of 5-HT, 5-methoxytryptamine (5-MeO-T), 1-(3 chlorophenyl) piperazine (mCPP) and 5-carboxamidotryptamine (5-CT) (10, 30, 100 and 300 microg kg(-1) each), produced dose-dependent tachycardic responses. Interestingly, DOI (10 - 1000 microg kg(-1), i.v.) induced only slight, dose unrelated, tachycardic responses, whilst the 5-HT(2C) agonist, Ro 60-0175 (10 - 1000 microg kg(-1), i.v.), produced a slight tachycardia only at 300 and 1000 microg kg(-1). In contrast, sumatriptan and 1-(m-trifluoromethylphenyl)- piperazine (TFMPP) were inactive. The rank order of potency was: 5-HT > or =5-MeO T> mCPP > or =5-CT > or =DOI > Ro 60-0175. 3. The tachycardic responses to 5-HT, which remained unaffected after i.v. saline (0.3 and 1 ml kg(-1)) or propranolol (3 mg kg(-1)), were selectively blocked by the 5-HT(2A) antagonists ketanserin (30 and 100 microg kg(-1)) or spiperone (10 and 30 microg kg(-1)) as well as by the non-selective 5-HT(2) antagonists, ritanserin (10 and 30 microg kg(-1)) or mesulergine (100 microg kg(-1)). Remarkably, these responses were unaffected by the antagonists rauwolscine (5-HT(2B)), SB204741 (5-HT(2B/2C)) or Ro 04-6790 (5 ht(6)) (300 and 1000 microg kg(-1) each). 4. These results suggest that the 'atypical' 5-HT(2) receptors mediating tachycardia in reserpinized pithed rats are pharmacologically similar to the 5-HT(2A) receptor subtype. PMID- 11906968 TI - Inhibition of cyclo-oxygenase-2 exacerbates ischaemia-induced acute myocardial dysfunction in the rabbit. AB - 1. The effects of treatment with a number of cyclo-oxygenase inhibitors, (celecoxib, meloxicam, DuP-697 and aspirin) on ischaemia-reperfusion-induced myocardial dysfunction were examined using an in vitro perfused rabbit heart model. 2. Ischaemia resulted in myocardial dysfunction, as indicated by a significant increase in left ventricular end diastolic pressure and marked changes in coronary perfusion pressure and left ventricular developed pressure. In the post-ischaemic state, coronary perfusion pressure increased dramatically, left ventricular developed pressure recovered to a small degree and there were significant increases in creatinine kinase release (indicative of myocardial damage) and prostacyclin release. 3. Pretreatment with aspirin, or with drugs that selectively inhibit cyclo-oxygenase-2 (celecoxib, meloxicam and DuP-697), resulted in a concentration-dependent exacerbation of the myocardial dysfunction and damage. Exacerbation of myocardial dysfunction and damage was evident with 10 microM concentrations of the cyclo-oxygenase-2 inhibitors, which inhibited prostacyclin release but did not affect cyclo-oxygenase-1 activity (as measured by whole blood thromboxane synthesis). 4. NCX-4016, a nitric oxide-releasing aspirin derivative, significantly reduced the myocardial dysfunction and damage caused by ischaemia and reperfusion. Beneficial effects were observed even at a concentration (100 microM) that significantly inhibited prostacyclin synthesis by the heart. 5. The results suggest that prostacyclin released by cardiac tissue in response to ischaemia and reperfusion is derived, at least in part, from cyclo oxygenase-2. Cyclo-oxygenase-2 plays an important protective role in a setting of ischaemia-reperfusion of the heart. PMID- 11906970 TI - NCX-701 (nitroparacetamol) is an effective antinociceptive agent in rat withdrawal reflexes and wind-up. AB - 1. Non-steroidal anti-inflammatory drugs (NSAIDs) are effective anti-inflammatory and analgesic drugs although they also induce unwanted side effects due to the inhibition of the physiological effects regulated by prostaglandins. This has led to the search for new compounds with fewer side effects, such as the nitro-NSAIDs (NO-NSAIDs). Paracetamol is an analgesic drug devoid of some of the side effect of the NSAIDs but without anti-inflammatory activity. NCX-701 is a nitric oxide releasing version of paracetamol with anti-inflammatory and analgesic properties. 2. We have tested, in the single motor unit technique, the antinociceptive actions of intravenous cumulative doses of NCX-701 vs paracetamol, studying their antinociceptive effects in responses to noxious mechanical and electrical stimulation (wind-up). 3. Paracetamol did not induce any significant effect at the doses tested (maximum of 480 micromol kg(-1), 72.5 mg kg(-1)). NCX-701 however was very effective in reducing responses to noxious mechanical stimulation (32+/-10% of control response) and wind-up (ED(50) of 147+/-1 micromol kg(-1), 41.5+/-0.3 mg kg(-1)). The inhibition was not reversed by 1 mg kg(-1) of the opioid antagonist naloxone. In control experiments performed with either the vehicle or the NO donor NOC-18, no significant changes were observed in the nociceptive responses studied. 4. We conclude that NCX-701 is a very effective non-opioid antinociceptive agent in normal animals and its action is located mainly at central areas. The antinociceptive effect was not due solely to the release of NO. PMID- 11906969 TI - The effects of neuroleptics on the GABA-induced Cl- current in rat dorsal root ganglion neurons: differences between some neuroleptics. AB - 1. Several neuroleptics inhibited the 3 microM gamma-aminobutyric acid induced chloride current (GABA-current) on dissociated rat dorsal root ganglion neurons in whole-cell patch-clamp investigations. 2. The IC(50) for clozapine, zotepine, olanzapine, risperidone and chlorpromazine were 6.95, 18.26, 20.30, 106.01 and 114.56 microM, respectively. The values for the inhibitory effects of neuroleptics on the GABA (3 microM)-current, which were calculated by the fitting Hill's equations where the concentrations represent the mean therapeutic blood concentrations, were ranked clozapine>zotepine>chlorpromazine>olanzapine>risperidone. These inhibitory effects, weighted with the therapeutic concentrations of neuroleptics, were correlated with the clinical incidences of seizure during treatment with neuroleptics. 3. Clozapine reduced the picrotoxin-inhibiton, and may compete with a ligand of the t-butylbicyclophosphorothionate (TBPS) binding site. 4. Haloperidol and quetiapine did not affect the peak amplitude of the GABA (3 microM)-current. However, haloperidol reduced the clozapine-inhibition, and may antagonize ligand binding to TBPS binding site. 5. Neuroleptics including haloperidol and quetiapine enhanced the desensitization of the GABA (3 microM) current. However, haloperidol and quetiapine at 100 microM inhibited the desensitization at the beginning of application. 6. Blonanserin (AD-5423) at 30 and 50 microM potentiated the GABA (3 microM)-current to 170.1+/-6.9 and 192.0+/ 10.6% of the control current, respectively. Blonanserin shifted GABA concentration-response curve leftward. Blonanserin only partly negatively interacted with diazepam. The blonanserin-potentiation was not reversed by flumazenil. Blonanserin is not a benzodiazepine receptor agonist. 7. The various effects of neuroleptics on the GABA-current may be related to the clinical effects including modifying the seizure threshold. PMID- 11906971 TI - The human 5-HT7 serotonin receptor splice variants: constitutive activity and inverse agonist effects. AB - 1. Using membranes from stably or transiently transfected HEK293 cells cultured in 5-HT-free medium and expressing the recombinant human 5-HT(7) receptor splice variants (h5-HT(7(a)), h5-HT(7(b)) and h5-HT(7(d))), we compared their abilities to constitutively activate adenylyl cyclase (AC). 2. All h5-HT(7) splice variants elevated basal and forskolin-stimulated AC. The basal AC activity was reduced by the 5-HT(7) antagonist methiothepin and this effect was blocked by mesulergine (neutral 5-HT(7) antagonist) indicating that the inhibitory effect of methiothepin is inverse agonism at the 5-HT(7) receptor. 3. Receptor density correlated poorly with constitutive AC activity in stable clonal cell lines and transiently transfected cells. Mean constitutive AC activity as a percentage of forskolin-stimulated AC was significantly higher for the h5-HT(7(b)) splice variant compared to the h5-HT(7(a)) and h5-HT(7(d)) splice variants but only in stable cell lines. 4. All eight 5-HT antagonists tested inhibited constitutive AC activity of all splice variants in a concentration-dependent manner. No differences in inverse agonist potencies (pIC(50)) were observed between the splice variants. The rank order of potencies was in agreement and highly correlated with antagonist potencies (pK(b)) determined by antagonism of 5-HT stimulated AC activity (methiothepin >metergoline> mesulergine > or = clozapine > or = spiperone > or = ritanserin > methysergide > ketanserin). 5. The efficacy of inverse agonism was not receptor level dependent and varied for several 5-HT antagonists between membrane preparations of transiently and stably transfected cells. 6. It is concluded that the h5-HT(7) splice variants display similar constitutive activity and inverse agonist properties. PMID- 11906974 TI - Research misconduct. PMID- 11906972 TI - Experimental model of escape phenomenon in hamsters and the effectiveness of YM 53601 in the model. AB - 1. The aim of this study was to establish an experimental model of the escape phenomenon, in which plasma cholesterol, initially reduced by a 3-hydroxy-3 methylglutaryl CoA (HMG-CoA) reductase inhibitor such as pravastatin, increases again on long-term administration. We also evaluated the efficacy of YM-53601 ((E)-2-[2-fluoro-2- (quinuclidin-3-ylidene) ethoxy]-9H-carbazole monohydrochloride), a squalene synthase inhibitor, in this model. 2. Pravastatin inhibited cholesterol biosynthesis in hamster primary hepatocytes (IC(50), 14 nM). After pre-treatment with pravastatin, in contrast, almost no effect on cholesterol biosynthesis was seen. 3. In hamsters fed a high fat diet, 3 mg kg( 1) pravastatin for 9 days decreased plasma non-HDL cholesterol (total cholesterol - high density lipoprotein cholesterol) (P<0.01), but this effect was lost between 17 and 27 days of treatment, accompanied by an increase in HMG-CoA reductase activity. No such increase in plasma non-HDL cholesterol was seen with YM-53601 at 30 mg kg(-1) after 9 (P<0.001), 17 (P<0.01) or 27 (P<0.001) days of treatment. Replacement of pravastatin with YM-53601 caused a decrease in plasma non-HDL cholesterol by 53% (P<0.001) and in HMG-CoA reductase activity. 4. This animal model thus satisfactorily replicates the escape phenomenon observed in humans and may therefore be useful in evaluation of lipid-lowering agents, specifically comparison of HMG-CoA reductase inhibitors. Further, YM-53601 may be useful in the treatment of hypercholesterolemia without induction of the escape phenomenon. PMID- 11906973 TI - Red wine polyphenols increase calcium in bovine aortic endothelial cells: a basis to elucidate signalling pathways leading to nitric oxide production. AB - 1. The present study investigates the mechanisms by which polyphenolic compounds from red wine elicit Ca(2+) mobilization in bovine aortic endothelial cells (BAECs). Two polyphenol-containing red wine extracts, red wine polyphenolic compounds (RWPC) and Provinols, and delphinidin, an anthocyanin were used. 2. RWPC stimulated a Ca(2+)-dependent release of nitric oxide (NO) from BAECs accounting for the relaxation of endothelium-denuded rat aortic rings as shown by cascade bioassay. 3. RWPC, Provinols and delphinidin increased cytosolic free calcium ([Ca(2+)](i)), by releasing Ca(2+) from intracellular stores and by increasing Ca(2+) entry. 4. The RWPC-induced increase in [Ca(2+)](i) was decreased by exposure to ryanodine (30 microM), whereas Provinols and delphinidin induced increases in [Ca(2+)](i) were decreased by bradykinin (0.1 microM) and thapsigargin (1 microM) pre-treatment. 5. RWPC, Provinols and delphinidin-induced increases in [Ca(2+)](i) were sensitive to inhibitors of phospholipase C (neomycin, 3 mM; U73122, 3 microM) and tyrosine kinase (herbimycin A, 1 microM). 6. RWPC, Provinols and delphinidin induced herbimycin A (1 microM)-sensitive tyrosine phosphorylation of several intracellular proteins. 7. Provinols released Ca(2+) via both a cholera (CTX) and pertussis toxins (PTX)-sensitive pathway, whereas delphinidin released Ca(2+) only via a PTX-sensitive mechanism. 8. Our data contribute in defining the mechanisms of endothelial NO production caused by wine polyphenols including the increase in [Ca(2+)](i) and the activation of tyrosine kinases. Furthermore, RWPC, Provinols and delphinidin display differences in the process leading to [Ca(2+)](i) increases in endothelial cells illustrating multiple cellular targets of natural dietary polyphenolic compounds. PMID- 11906975 TI - How are patients informed about their HIV test results? AB - BACKGROUND: AIDS and HIV are hot topics in public health nowadays, but little information is available about the way in which HIV test results are communicated to patients. OBJECTIVE: Our aim was to find out about the way in which patients are informed about their HIV test results and the delay they experience. METHOD: Since 1996, a representative network of sentinel GPs has recorded data about requests for HIV tests, risk factors and the way in which and the delay with which patients are informed about their HIV test results. RESULTS: Between 1996 and 1999, 4807 requests for an HIV test were recorded. Test results were given mostly by phone (41.9%). Patients at risk were informed more often during a planned follow-up. Anxious patients were informed more often about the results by phone, and in 61.2% test results were communicated during the first week following the test. Results were given earlier if patients were anxious or had themselves asked for the test. Although non-consensual HIV testing is against the European guidelines on informed consent, 102 tests (2.2%) were performed without informed consent. CONCLUSIONS: Even though notification by phone decreases the delay, physicians should be encouraged to make follow-up appointments to inform the patient about the test results. A face-to-face conversation is the only way in which physicians can offer valuable post-test counselling. Physicians should be informed about the unlawfulness of non-consensual HIV testing. PMID- 11906976 TI - Euthanasia revisited. AB - Euthanasia is a debatable issue. It is illegal all over the world. The Netherlands is the only country where euthanasia and physician-assisted suicide are openly practised since the physician performing these acts will not be prosecuted under certain circumstances. There were several court cases and court decisions that affected the development of euthanasia and physician-assisted suicide in individual countries. When a patient asked for euthanasia, it was very important to find out the underlying reasons and make all legal means available to relieve the pain and other distressing symptoms. PMID- 11906977 TI - Ethics of qualitative research: are there special issues for health services research? AB - BACKGROUND: An increasing volume of qualitative research and articles about qualitative methods has been published recently in medical journals. However, compared with the extensive debate in social sciences literature, there has been little consideration in medical journals of the ethical issues surrounding qualitative research. A possible explanation for this lack of discussion is that it is assumed commonly that qualitative research is unlikely to cause significant harm to participants. There are no agreed guidelines for judging the ethics of qualitative research proposals and there is some evidence that medical research ethics committees have difficulty making these judgements. OBJECTIVES: Our aim was to consider the ethical issues which arise when planning and carrying out qualitative research into health and health care, and to offer a framework within which health services researchers can consider these issues. RESULTS: Four potential risks to research participants are discussed: anxiety and distress; exploitation; misrepresentation; and identification of the participant in published papers, by themselves or others. Recommended strategies for reducing the risk of harm include ensuring scientific soundness, organizing follow-up care where appropriate, considering obtaining consent as a process, ensuring confidentiality and taking a reflexive stance towards analysis. CONCLUSIONS: While recognizing the reservations held about strict ethical guidelines for qualitative research, we argue for further debate of these issues so that the health services research community can move towards the adoption of agreed standards of good practice. In addition, we suggest that empirical research is desirable in order to quantify the actual risks to participants in qualitative studies. PMID- 11906978 TI - Whose autonomy? Which choice? A study of GPs' attitudes towards patient autonomy in the management of low back pain. AB - BACKGROUND: Respect for patient autonomy is an important ethical principle for medical practitioners; however, previous investigators have reported inconsistent attitudes amongst practitioners towards respect for patient autonomy. This study in empirical ethics used qualitative methods to investigate GPs' attitudes towards respect for patient autonomy in consultations for low back pain. OBJECTIVES: The aim of this study was to explore GPs' attitudes towards respect for patient autonomy by analysing attitudes towards four issues in the management of low back pain which raise ethical and practical dilemmas. METHODS: Participants were 21 GPs selected from general practice in South Australia by stratified, purposive sampling aimed at maximizing diversity. Semi-structured interviews were tape-recorded, transcribed and analysed using codes developed from the transcripts, with additional theoretical codes. In the analysis, attitudes towards patient autonomy in the four issues were characterized as autonomy-respecting, intermediate or controlling. RESULTS: The results showed individual inconsistencies in GPs' attitudes towards respect for patient autonomy. For example, the majority of GPs accepted patient autonomy in the use of complementary therapies, but were controlling with regard to the use of analgesics. Attitudes to duration of time off work were spread evenly, whilst controlling attitudes towards use of X-rays were modified by patient requests for X-rays. CONCLUSIONS: These results suggest that GP attitudes towards patient autonomy are modified by ethical and pragmatic factors, and vary depending upon the nature of the issue in question. PMID- 11906979 TI - Preferences for gender of family physician among Canadian European-descent and South-Asian immigrant women. AB - OBJECTIVE: The aim of this study was to investigate expressed preferences for family physician (FP) gender among Canadian European-descent (CED) and Canadian South-Asian (CSA) immigrant women. METHOD: An 'on-site' survey was conducted in community-based institutions in Toronto in order to determine preferences for the gender of FP under various health care scenarios: overall health care; gender sensitive examinations; emotional problems; general ailments; and life threatening conditions. RESULTS: Ninety-four women responded to this survey (CED = 50, CSA = 44), response rate 77.3%. For all health care scenarios, CED and CSA women similarly expressed either a preference for a female FP or no preference. More than two-thirds of women preferred a female FP for gynaecological examinations (CED, 72.9%; CSA, 83.7%) or examinations with private body part exposure (CED, 72%; CSA, 81.8%). For 'emotional problems', half of the women preferred a female FP and the other half had no preference. A similar pattern was observed for 'overall health care', with some shift to female physician preference among CSA women (60.5%) compared with CED women (53.2%). For the 'overall health care' scenario, CED and CSA women who preferred a female FP had a higher frequency of seeing female physicians within the last 5 years (CED, P < or = 0.01; CSA, P < or = 0.05), and attributed 'positive' social skills more to female physicians (CED, P < or = 0.01; CSA, P < or = 0.01) compared with women with no preference for the gender of the FP. Yet, CED women with a female FP preference were more likely to have a concurrent female FP (P < or = 0.01), and to rate past experiences with female physicians as high (P < or = 0.01) and with male physicians as low (P < or = 0.05) compared with CED women with no preference. In the CSA group, women with a preference for a female FP were more likely to be unemployed (P < or = 0.01) and have low social support (P < or = 0.01). CONCLUSIONS: Despite similar physician gender preference patterns, factors associated with these preferences show some differences between CED and CSA women. PMID- 11906980 TI - A new method for describing smokers' consulting behaviours which indicate their motivation to stop smoking: an exploration of validity and reliability. AB - BACKGROUND: Smokers vary in their readiness to try stopping smoking, but there currently are no objective tools for identifying smokers' consulting behaviours which indicate their level of motivation to try stopping smoking. OBJECTIVE: The aim of this study was to investigate the construct validity and inter-observer reliability of the Smokers' Motivation Code (SMC). METHODS: General practice consultations between 29 different Leicestershire GPs and their patients were video-recorded. In 47 consultations, regular or occasional smokers discussed smoking with their GPs and their consulting behaviour was coded using the SMC. The reliability of three different observers' codings was investigated. Construct validity was also investigated by comparing smokers' consulting behaviours coded using the SMC with measures of motivation to stop smoking recorded on pre consultation questionnaires. RESULTS: Two pairs of observers achieved good reliability when using the SMC to code smokers' consulting behaviours during a subset of 11 video-recorded consultations. For readiness behaviours (indicating motivation to stop smoking), kappas were 0.82 and 0.65, and for resistance behaviours (indicating little motivation to stop smoking), kappas were 0.74 and 1.0. For the 37 consultations attended by regular smokers, complete pre consultation questionnaires were obtained. Smokers displaying readiness behaviours were significantly more likely than others to report having tried to stop in the past year, thinking about or trying to stop and to agree that their health would improve if they stopped smoking. Smokers displaying resistant behaviours were significantly less likely to report thinking about stopping/trying to stop smoking. CONCLUSION: We have provided some evidence to support the construct validity and inter-observer reliability of the SMC and have identified some consulting behaviours which might indicate smokers' motivation to stop smoking. Further work is needed to determine whether smokers' consulting behaviour can be used to predict future quit attempts. PMID- 11906981 TI - Effects of guided written disclosure of stressful experiences on clinic visits and symptoms in frequent clinic attenders. AB - BACKGROUND: Psychosocial variables such as major stressful life events/daily stressful events have been associated with health care utilization. OBJECTIVE: Our aim was to examine the effects of a guided disclosure protocol (GDP) of past traumas on symptoms and clinic visits among frequent clinic attenders. METHODS: Forty-one frequent clinic attenders (> or =2 visits/3 months) took part. Patients were randomly assigned individually to either a casual content writing control group (n = 19) or a trauma content writing experimental GDP group (n = 22). GDP patients wrote about an upsetting event chronologically (day 1), verbally described their thoughts and feelings and described the event's impact on life (day 2), and finally wrote about their current perspective on and future coping with the event (day 3). Three months later, patients were reassessed blindly for symptoms and clinic visits, and an average of 15 months later they were assessed blindly for clinic visits again. RESULTS: Compared with controls, GDP patients reported lower symptom levels at 3 months (2.3 versus 5.2), and made fewer clinic visits during the 3 (1.3 versus 3.0) and 15 month (5.1 versus 9.7) follow-ups. The percentage of GDP patients making > or =10 visits during the 15 month follow up was smaller (10%) than among controls (33%). CONCLUSIONS: The findings extend previous findings to frequent clinic users, using a new form of written disclosure aimed at shifting trauma from implicit to explicit memory. The GDP may be an inexpensive additional intervention in primary care for reducing symptoms and clinic visits among frequent clinic users. PMID- 11906982 TI - The patient as a source to improve the medical record. AB - BACKGROUND: The problem list is an important tool in general practice for care as well as research purposes. As the central part of the problem-oriented medical record, it lists the main medical problems which the GP wants to have knowledge of during any patient encounter. The assessment of its quality is usually made by comparing with other sources of information on the patient's problems. OBJECTIVE: This study addresses the question of to what extent the problem list can be improved by asking the patient about their own medical problems. METHODS: During 7 weeks, all patients who visited three GPs in a health care centre in an Amsterdam suburb were interviewed. During the interview, they were confronted with the problem list made by their own GP and stimulated to make suggestions for addition or removal of problems. RESULTS: All in all, patients were in agreement with 88% of all listed problems. The completeness of the problem list could be increased by 28%, while 4% ultimately were removed: a net gain of 24%. CONCLUSION: The patient can be used as a sourcetool for improvement of the quality of the problem list when its prime function is patient care. It becomes more complicated when the problem list also serves a research purpose. Clear inclusion rules will then have to be formulated. PMID- 11906983 TI - Lay versus professional motivation for asthma treatment: a cross-sectional, qualitative study in a single Glasgow general practice. AB - OBJECTIVE: Our aim was to identify the factors which motivate patient self management in asthma and to compare the results with the factors which appear to have motivated the content of the British Thoracic Society (BTS) clinical guidelines for chronic asthma in adults. METHOD: We conducted a cross-sectional, qualitative survey of asthma patients from a single general practice list in Glasgow, Scotland. Twenty-three adult asthma patients on at least step 2 of the BTS guidelines were selected from the practice asthma register. RESULTS: Only seven of the 23 subjects had asthma treatment goals. People with asthma are motivated by the effect of the illness on self-image, the experience of symptoms, the value of life experiences affected, the perceived consequences of asthma, their acceptance of the diagnosis and their attitude towards medication. Asthma is largely viewed as unproblematic. CONCLUSIONS: The BTS guidelines appear to be motivated by a desire to heighten professional awareness about the disease. Patient goals and preferences for asthma treatment are largely unrecognized by the guidelines. A concordant model of disease management, involving the explicit acknowledgement of patient goals by professionals, alongside their own goals for treatment, may improve adherence to treatment perceived by patients as relevant and achievable. PMID- 11906984 TI - Medically unexplained symptoms and the problem of power in the primary care consultation: a qualitative study. AB - BACKGROUND: Patients presenting in primary care frequently exhibit physical symptoms that may be unrelated to organic pathology. Such symptoms are commonly regarded as products of psychological or emotional problems, and their legitimacy as 'medical' matters is often called into question. OBJECTIVES: Our aim was to explore GPs' attitudes to the management of patients that present with medically unexplained symptoms in primary care. METHODS: Semi-structured interviews were conducted with 15 GPs in North-West England. Interviews were audio-taped and subsequently transcribed and analysed using a constant comparison technique. RESULTS: Subjects conceptualized patients presenting with medically unexplained symptoms as the presentation of psychological distress. They presented problems of control and authority in the consultation, and difficulties in managing this had a negative impact on the doctor-patient relationship. Such consultations were frustrating for the GP and potentially harmful to the patient. CONCLUSION: Patients with medically unexplained symptoms were seen to be presenting with inappropriate symptoms that were a manifestation of emotional or social distress. GPs felt ill-equipped to deal with the presentations and the frustrations they felt and may need help in actively and productively managing these patients. PMID- 11906985 TI - Patients' views and feelings on the community-based teaching of undergraduate medical students: a qualitative study. AB - BACKGROUND: The 1993 directive Tomorrow's Doctors recommended an increase in community-based teaching. In response, many new programmes have been established focusing on the teaching of clinical skills to pairs or groups of students in general practice, when patients are asked to see the students in the practice or in their homes, specifically to assist with teaching. This differs from the traditional model of teaching primary care, when one student sits with the doctor while s/he consults. Although current research suggests that patients are happy for one student to be present during a consultation with their GP, little or nothing is known about their views of this new method of teaching basic clinical skills in the community. If the new community-based teaching programmes are to be sustainable, continuing support from the patients is necessary. Students have been taught clinical skills in the community at University College London Medical School for several years. Research has demonstrated its effectiveness and its popularity with students. However, to date, patients' views have not been explored. OBJECTIVE: Our aim was to determine the patients' views and feelings on community-based teaching, in order to discover both the positive and negative aspects for participating patients. METHOD: A qualitative semi-structured interview study was carried out in undergraduate teaching general practices in North London. RESULTS: Respondents felt very positive about participating in the community-based teaching programme. There were two underlying components to this: altruism and personal gain. Within altruism, reasons included: provision of a service to the community and repaying the system. Aspects of personal gain included: improved knowledge, improved self-esteem and companionship. Patient concerns included: embarrassment, reinforcement of the sick role and concerns about student access to notes. CONCLUSIONS: Patients enjoy their involvement in community-based teaching and perceive themselves as making a valuable contribution. The findings of the research will be reassuring for doctors who presently are involved and those who plan to be involved in the future. Doctors need to be aware of the possible shifts in the doctor-patient relationship when actively seeking patients' help in the teaching. PMID- 11906986 TI - Prevention of cervical cancer in a poor population in Brazil. AB - OBJECTIVE: The purpose of this study was to determine the possibility of providing a cervical screening facility to a poor population. METHODS: A cross sectional study was conducted in the period from December 1994 to December 1997, with 1384 women from the poor districts of Barretos, Sao Paulo and three other neighbouring cities. Performed by a nurse, the programme included door-to-door interviews and cervical screening. The Papanicolaou smears were taken either at the community centre or at home using a portable gynaecological table transportable by bicycle, developed by the Institution. RESULTS: From 1384 interviewed women, 1044 (75.4%) agreed to undergo the examination and 499 (47.8%) had never had the test or had not had it repeated within the last 3 years. Among 1044 examined women, seven cases of carcinoma 'in situ', one invasive squamous cell carcinoma (stage IB) and two polyps were found. CONCLUSIONS: This study shows that programmes of cancer prevention in poor populations can be as successful as those carried out in more developed countries by taking advantage of innovations in the delivery of care. PMID- 11906987 TI - The genetics liaison nurse role as a means of educating and supporting primary care professionals. AB - BACKGROUND: Previous research with primary health care professionals has demonstrated consistently that education, training and support are necessary before there should be any expansion in primary care genetics. The genetic liaison nurse role has been suggested as one means of providing this education and support. OBJECTIVE: The aim of this study was to evaluate GP responses to the genetics liaison nurse role as a means of supporting community-based genetics services. METHODS: A self-completion postal questionnaire in primary care was sent to GPs working in Nottingham. Main outcome measures were assessment of potential usage of a genetic outreach professional in terms of time, roles and support for a pilot scheme RESULTS: A total of 182 (55.0%) of 331 GPs working in Nottingham returned a questionnaire. Although 54% did not believe that the genetics liaison nurse role would be useful in the present, most believed that such a role would definitely or probably (64%) be useful in the future. The most valued contribution was as a source of advice when genetics problems arise in a consultation. Providing education on specific genetic disorders and on clinical skills relevant to genetics were also seen as important. Many GPs would also use a liaison nurse to see patients prior to their attending an out-patient clinic with a clinical geneticist. Respondents suggested that each nurse should spend approximately 3 hours a month in each practice and be attached to between 10 and 20 practices. CONCLUSIONS: GPs appreciate that there may be limited genetics services provided in primary care at present, but this is likely to change in the near future. The genetics liaison nurse role should be evaluated as a means of providing genetics specialist outreach support for service delivery and to facilitate education. PMID- 11906988 TI - Primary care reform: a pilot study to test the evaluative potential of the Patient Enablement Instrument in Poland. AB - BACKGROUND: Primary health care reform is underpinned by a move towards patient centred holistic care. This pilot study uses the Patient Enablement Instrument (PEI) to assess outcome at a fundamental level: that of the patient and their doctor at consultation. OBJECTIVES: Our aim was to assess the evaluative potential of the PEI in relation to a reform programme in Poland by (i) comparing the outcomes of consultations (using the PEI) carried out by nine doctors (three diploma GPs who had participated in the training programme, three GPs who had not participated in the training programme and three polyclinic internists); and (ii) relating PEI scores to a proxy quality process measure (consultation length). METHODS: A cross-sectional quantitative questionnaire survey was carried out using the PEI. The subjects were patients consulting with nine doctors distributed within a single region around Gdansk. RESULTS: The overall results with the PEI and consultation length reflected UK experience. In addition, there were significant differences between groups in this pilot study. Patients seen by diploma GPs achieved higher patient enablement scores (mean 4.33, 95% confidence interval 4.09-4.58) relative to GPs (mean 3.44, 3.21-3.67) and polyclinic doctors (mean 3.23, 2.99-3.47). However, there is evidence of appreciable between-doctor variation in PEI scores within groups. The difference in patient enablement between groups was not affected by patient case mix, in contrast to the duration of consultation, which was. Holistically trained diploma GPs spent longer with patients with psychological problems. Patients seen by diploma GPs received longer consultations (mean 12.65 min, 95% confidence interval 12.18-13.13) relative to their colleagues (the GPs' mean was 10.11, 9.82-10.41 min; that of the polyclinic internists was 10.16, 9.81-10.50 min). The duration of consultation was positively correlated with patient enablement. CONCLUSION: The results of such training courses should be examined from the perspective of both the patient and their doctor. Significant differences were found in both patient enablement and consultation length between patients attending groups of doctors delivering primary care, but working from different paradigms. This pilot shows promising results which, if repeated in a larger study, would provide an objective means of evaluating such reform programmes. PMID- 11906989 TI - Walk-in clinics: patient expectations and family physician availability. AB - BACKGROUND: For over two decades, there has been controversy over the role and impact of walk-in clinics on primary health care. This study evaluates the providers' perspective on this topic. OBJECTIVE: The purpose of this qualitative study was to explore the perceptions and experiences of family physicians, emergency physicians and walk-in clinic physicians regarding the impact of walk in clinics on Ontario's health care system. METHODS: The qualitative method of focus groups was used in this study. There were nine focus groups, each consisting of 4-9 participants, with a total of 63 physicians. The different practitioners (family physicians, emergency physicians, walk-in clinic physicians) attended separate focus groups. The focus groups explored the physicians' perceptions and experiences regarding the role and impact of walk-in clinics on Ontario's health care system. The focus groups were audio-taped and transcribed verbatim. The qualitative data analysis program NUD*IST was used to organize the data during the sequential thematic analysis. RESULTS: Factors contributing to the growth and evolution of walk-in clinics in Ontario were identified. These included a perceived increase in patients' expectations for convenient health care and a perceived decrease in the availability of family physicians. These factors created a gap in primary care which was filled by walk in clinics. CONCLUSIONS: Participants' recommendations for narrowing this gap included an increase in both physician and patient accountability and changes to the current structure of primary health care delivery. These recommendations would either integrate walk-in clinics into the health care system or result in their elimination. PMID- 11906990 TI - The relevance of continuity of care: a solution for the chaos in the emergency services. AB - BACKGROUND: In Brazil, there continues to be an excessive use of emergency services by patients with elective medical problems. Those patients who report having a primary care physician are less likely to utilize the emergency department for non-urgent consultations. OBJECTIVE: The objective of this study was to compare patients who have a primary care physician with those who do not in relation to severity of their chief complaint at presentation in the emergency department. METHODS: The study was carried out as a cross-sectional interview based survey at the Conceicao Hospital Emergency Service in Porto Alegre (Brazil). The sample was 553 patients selected through a systematic random sampling, and the response rate was 88%. The data entry and analysis were performed using the software Epi-info, EGRET and SPSS. The analysis included simple statistics to determine the prevalence of the conditions being investigated and the effect of independent variables (regular doctor) in relation to the dependent variable (severity of disease) through logistic regression. RESULTS: The chief complaints were divided up as follows: 15% emergency cases, 46% urgent cases and 39% elective. The chief complaint was defined as urgent or emergency if it exhibited a significantly statistical association with the following independent variables, after being analysed by a logistic regression model: patients who reported having a primary care physician [odds ratio (OR) = 2.98, 95% confidence interval (CI) = 1.84-4.80] and patients who usually go to the emergency room by car (OR = 2.67, 95% CI = 1.75-4.05). CONCLUSION: One strategy to reduce the number of non-urgent consultations at emergency rooms is to establish a close out-patient relationship between patients and physician. There is a need to optimize the health care of patients who have non-urgent problems but still seek the emergency department through strategies at the primary health care level-especially when continuous care is available-and where a comprehensive approach with an emphasis on prevention would stimulate better quality of care at a lower cost. PMID- 11906991 TI - Teenage pregnancy: whose problem is it? PMID- 11906992 TI - Rapamycin eluting stent: the onset of a new era in interventional cardiology. PMID- 11906993 TI - Fibrinolytic treatment for elderly patients with acute myocardial infarction. PMID- 11906994 TI - Thrombolysis: too old and too young. PMID- 11906995 TI - Natriuretic peptide receptors and the heart. PMID- 11906996 TI - Myocardial infarction. PMID- 11906997 TI - Cardiac hypertrophy and oxidative stress: a leap of faith or stark reality? PMID- 11906999 TI - Myocardial viability by contrast enhanced MRI in a patient with left bundle branch block showing a severe defect on FDG-PET. PMID- 11907000 TI - Vasomotion and nitric oxide bioactivity in diseased coronary arteries. AB - Atheromatous coronary stenoses are no longer considered to be passive structures, instead having the capacity for dynamic, often transient, change which may take the form of constriction or dilatation in response to either endogenous or external stimulation. PMID- 11907002 TI - Single large metastatic tumor growing progressively and occupying right ventricular cavity. PMID- 11907001 TI - Effect of multisite pacing on ventricular coordination. AB - OBJECTIVE: To determine the effect of multisite pacing on left ventricular function. DESIGN: Prospective observational study. PATIENTS: 18 patients with heart failure with a dilated poorly functioning left ventricle (LV) and left bundle branch block. INTERVENTIONS: Pacing for 5 minutes in random order at the right ventricle (RV) apex, RV outflow tract, mid posterolateral LV, RV apex and LV simultaneously, and RV outflow tract and LV simultaneously. The best achieved measurements with RV, LV, and biventricular pacing were compared. MAIN OUTCOME MEASURES: LV dimension, filling characteristics, and long axis indices were measured on echocardiography simultaneously with LV pressure. Cycle efficiency (%)--that is, the ratio of the area of the acquired pressure dimension loop to that of the ideal loop for that segment--quantified coordination. RESULTS: The pacing site that gave the best achieved cycle efficiency differed between patients (biventricular in five, LV in two, RV in seven, and no site in four). In patients with baseline incoordination (cycle efficiency < or = 72%, n = 12) cycle efficiency improved significantly with RV pacing (cycle efficiency 76%, p = 0.01) but not with LV (65%) or biventricular (67%) pacing. LV based pacing induced premature short axis contraction in a subset of patients (n = 4), which was associated with a prolonged time from the Q wave on the ECG to the onset of inward movement of the long axis (from apex to mitral ring): biventricular 145 ms, LV 105 ms, RV 85 ms (biventricular v RV, p < 0.05). Excluding patients with baseline incoordination in whom premature activation occurred, pacing at all sites led to a similar increase in cycle efficiency (RV 78%, LV 72%, biventricular 73%). CONCLUSIONS: Ventricular coordination can be improved with pacing in patients with baseline incoordination. Short and long axis fibres may be asynchronised in a subset of patients with LV or biventricular pacing, which may worsen coordination. The clinical significance of these findings remains to be defined. PMID- 11907004 TI - Anomalous left main stem stenosis. PMID- 11907003 TI - Dobutamine stress echocardiography for the detection of myocardial viability in patients with left ventricular dysfunction taking beta blockers: accuracy and optimal dose. AB - OBJECTIVE: To assess the accuracy of dobutamine stress echocardiography (DSE) and the optimal dose of dobutamine to detect myocardial viability in patients with ischaemic left ventricular (LV) dysfunction who are taking beta blockers, using the recovery of function six months artery revascularisation as the benchmark. PATIENTS: 17 patients with ischaemic LV dysfunction (ejection fraction < 40%) and chronic treatment with beta blockers scheduled to undergo surgical revascularisation. SETTING: Regional cardiothoracic centre. METHODS: All patients underwent DSE one week before and resting echocardiography six months after revascularisation. A wall motion score was assigned to each segment for each dobutamine infusion stage, using the standard 16 segment model of the left ventricle. The accuracy of DSE to predict recovery of resting segmental function was calculated for low dose (5 and 10 microg/kg/min) and for a full protocol of dobutamine infusion (5 to 40 microg/kg/min). RESULTS: Of the 272 segments studied, 158 (58%) were dysfunctional at rest, of which 79 (50%) improved at DSE and 74 (47%) recovered resting function after revascularisation. Analysis of results with a low dose showed a significantly lower sensitivity and negative predictive value than with a full protocol (47% v 81%, p < 0.001 and 65% v 82%, p < 0.05, respectively). The accuracy in the full protocol analysis was comparable with that reported in patients no longer taking beta blockers but was significantly lower than that in the low dose analysis (78% v 66%, p < 0.001). CONCLUSIONS: Findings suggest that beta blocker withdrawal is not necessary before DSE when viability is the clinical information in question. However, a completed protocol with continuous image recording is required to detect the full extent of viability. PMID- 11907006 TI - Covered stent for iatrogenic coronary arteriovenous fistula in heart transplant recipient. PMID- 11907005 TI - Is Doppler tissue velocity during early left ventricular filling preload independent? AB - BACKGROUND: Transmitral Doppler flow indices are used to evaluate diastolic function. Recently, velocities measured by Doppler tissue imaging have been used as an index of left ventricular relaxation. OBJECTIVE: To determine whether Doppler tissue velocities are influenced by alterations in preload. METHODS: Left ventricular preload was altered in 17 patients (all men, mean (SD) age, 49 (8) years) during echocardiographic measurements of left ventricular end diastolic volume, maximum left atrial area, peak early Doppler filling velocity, and left ventricular myocardial velocities during early filling. Preload altering manoeuvres included Trendelenberg (stage 1), reverse Trendelenberg (stage 2), and amyl nitrate (stage 3). Systolic blood pressure was measured at each stage. RESULTS: In comparison with baseline, left ventricular end diastolic volume (p = 0.001), left atrial area (p = 0.003), peak early mitral Doppler filling velocity (p = 0.01), and systolic blood pressures (p = 0.001) were all changed by preload altering manoeuvres. Only left ventricular myocardial velocity during early filling remained unchanged by these manoeuvres. CONCLUSIONS: In contrast to standard transmitral Doppler filling indices, Doppler tissue early diastolic velocities are not significantly affected by physiological manoeuvres that alter preload. Thus Doppler tissue velocities during early left ventricular diastole may provide a better index of diastolic function in cardiac patients by providing a preload independent assessment of left ventricular filling. PMID- 11907007 TI - Oral beraprost sodium improves exercise capacity and ventilatory efficiency in patients with primary or thromboembolic pulmonary hypertension. AB - OBJECTIVE: To investigate the effect of beraprost sodium, an orally active prostacyclin analogue, on exercise capacity and ventilatory efficiency in patients with primary pulmonary hypertension and chronic thromboembolic pulmonary hypertension. PATIENTS AND DESIGN: Symptom limited cardiopulmonary exercise testing was performed before and 3 (1) months (mean (SEM)) after beraprost treatment in 30 patients with precapillary pulmonary hypertension (14 with primary pulmonary hypertension and 16 with chronic thromboembolic pulmonary hypertension). RESULTS: Long term treatment with beraprost resulted in significant increases (mean (SEM)) in peak workload (87 (4) W to 97 (5) W, p < 0.001) and peak oxygen consumption (peak VO2, 14.9 (0.7) ml/kg/min to 16.8 (0.7) ml/kg/min, p < 0.001). Beraprost decreased the ventilatory response to carbon dioxide production during exercise (VE-VCO2 slope, 42 (2) to 37 (1), p < 0.001). No significant difference in the responses of these variables to beraprost treatment was observed between patients with primary pulmonary hypertension and chronic thromboembolic pulmonary hypertension. CONCLUSIONS: Oral administration of beraprost sodium may improve exercise capacity and ventilatory efficiency in patients with both primary and chronic thromboembolic pulmonary hypertension. PMID- 11907009 TI - Idebenone and reduced cardiac hypertrophy in Friedreich's ataxia. AB - BACKGROUND: Friedreich's ataxia encodes a protein of unknown function, frataxin. The loss of frataxin is caused by a large GAA trinucleotide expansion in the first intron of the gene, resulting in deficiency of a Krebs cycle enzyme, aconitase, and of three mitochondrial respiratory chain complexes (I-III). This causes oxidative stress. Idebenone, a short chain quinone acting as an antioxidant, has been shown to protect heart muscle against oxidative stress in some patients. OBJECTIVE: To assess the efficiency of idebenone on cardiac hypertrophy in Friedreich's ataxia. DESIGN: Prospective, open trial. SETTING: Tertiary care centre. METHODS: Idebenone (5 mg/kg/day) was given orally to 38 patients with Friedreich's ataxia aged 4-22 years (20 males, 18 females). Cardiac ultrasound indices were recorded before and after idebenone treatment. RESULTS: After six months, cardiac ultrasound indicated a reduction in left ventricular mass of more than 20% in about half the patients (p < 0.001). The shortening fraction was initially reduced in six of the 38 patients (by between 11-26%) and it improved in five of these. In one patient, the shortening fraction only responded to 10 mg/kg/day of idebenone. No correlation was found between responsiveness to idebenone and age, sex, initial ultrasound indices, or the number of GAA repeats in the frataxin gene. CONCLUSIONS: Idebenone is effective at controlling cardiac hypertrophy in Friedreich's ataxia. As the drug has no serious side effects, there is a good case for giving it continuously in a dose of 5-10 mg/kg/day in patients with Friedreich's ataxia at the onset of hypertrophic cardiomyopathy. PMID- 11907010 TI - Myocardial blood volume and the amount of viable myocardium early after mechanical reperfusion of acute myocardial infarction: prospective study using venous contrast echocardiography. AB - BACKGROUND: Myocardial capillary perfusion is a prerequisite of myocellular viability after reperfusion of acute myocardial infarction. It was hypothesised that the magnitude of myocardial capillary perfusion, assessed by transmural signal intensity in venous contrast echocardiography as a corollary of the blood volume of myocardial capillaries, and the amount of viable myocardium, represented by differential levels of contractile function two weeks after reperfusion, are correlated. OBJECTIVES: To evaluate the role of venous contrast echocardiography for the identification of viable myocardium in patients with acute myocardial infarction early after successful mechanical reperfusion. METHODS: 60 patients with a first acute myocardial infarction underwent venous contrast echocardiography several hours after successful mechanical reperfusion (median time interval 190 min.). The relative transmural videointensity (median (25th, 75th percentiles)) of akinetic segments was determined. After two weeks, contractile function was re-evaluated at rest and during dobutamine infusion if segments without functional recovery were present. RESULTS: Relative videointensity early after reperfusion differed significantly between functional groups after two weeks: normokinesia (88% (77%, 100%)), hypokinesia (74% (54%, 99%)), and akinesia with (61% (48%, 76%)) and without contractile reserve (31% (22%, 46%)). Relative videointensity and contractile function were significantly correlated (r = -0.67). The diagnostic accuracy of relative videointensity > 50% for prediction of contractility of initially akinetic segments at rest or during dobutamine was 82% (chi2 = 76.2, p < 0.001). CONCLUSIONS: Early after successful mechanical reperfusion of acute myocardial infarction, the magnitude of capillary perfusion in the perfusion territory of an infarct related artery is correlated with the amount of viable myocardium. Quantitative venous contrast echocardiography can be used for accurate identification of viable myocardium. PMID- 11907012 TI - Interception of aortic regurgitation by vegetation plug in a patient with infective endocarditis. PMID- 11907011 TI - Health related quality of life and health status in adult survivors with previously operated complex congenital heart disease. AB - OBJECTIVE: To examine the impact of previously operated complex congenital heart disease on health related quality of life and subjective health status and to determine the relation between these parameters and physical status. DESIGN: Cross sectional; information on medical follow up was sought retrospectively. SETTING: Patients were randomly selected from the archives of the paediatric cardiology department, Leiden University Medical Centre, Leiden, The Netherlands, and approached irrespective of current cardiac care or hospital of follow up. PATIENTS: Seventy eight patients with previously operated complex congenital heart disease (now aged 18-32 years) were compared with the general population. MAIN OUTCOME MEASURES: Health related quality of life was determined with a specifically developed questionnaire (Netherlands Organisation for Applied Scientific Research Academic Medical Centre (TNO-AZL) adult quality of life (TAAQOL)) and subjective health status was assessed with the 36 item short form health survey (SF-36). Physical status was determined with the objective physical index, Somerville index, and New York Heart Association functional class. RESULTS: Health related quality of life of the patients was significantly worse than that of the general population in the dimensions gross motor functioning and vitality (p < 0.01). Correlations between health related quality of life and physical status were poor. Patients had significantly worse subjective health status than the general population in the dimensions physical functioning, role functioning physical, vitality, and general health perceptions (p < 0.01). Correlations between subjective health status and physical indices were weak. CONCLUSION: Adult survivors with previously operated complex congenital heart disease experienced limitations only in the physical dimensions of health related quality of life and subjective health status. Objectively measured medical variables were only weakly related to health related quality of life. These results indicate that, when evaluating health related quality of life, dedicated questionnaires such as the TAAQOL should be used. PMID- 11907014 TI - Progressive cardiac hypertrophy and dysfunction in atrial natriuretic peptide receptor (GC-A) deficient mice. AB - OBJECTIVE: To investigate how permanent inhibition of guanylyl cyclase A receptor (GC-A) affects cardiac function. METHODS: Hearts of GC-A-/- and corresponding wild type mice (GC-A+/+) were characterised by histological, western blotting, and northern blotting analyses. Cardiac function was evaluated in isolated, working heart preparations. RESULTS: At 4 months of age, GC-A-/- mice had global cardiac hypertrophy (about a 40% increase in cardiac weight) without interstitial fibrosis. Examination of heart function found a significant delay in the time of relaxation; all other parameters of cardiac contractility were similar to those in wild type mice. At 12 months, the hypertrophic changes were much more severe (about a 61% increase in cardiac weight), together with a shift in cardiac gene expression (enhanced concentrations of atrial natriuretic peptide (3.8-fold), B type natriuretic peptide (2-fold), beta myosin heavy chain (1.6-fold) and alpha skeletal actin (1.7-fold) mRNA), increased expression of cytoskeletal tubulin and desmin (by 29.6% and 25.6%, respectively), and pronounced interstitial fibrosis. These changes were associated with significantly impaired cardiac contractility (+dP/dt decreased by about 10%) and relaxation (-dP/dt decreased by 21%), as well as depressed contractile responses to pressure load (all p < 0.05). CONCLUSIONS: Chronic hypertension in GC-A-/- mice is associated with progressive cardiac changes--namely, initially compensated cardiomyocyte hypertrophy, which is complicated by interstitial fibrosis and impaired cardiac contractility at later stages. PMID- 11907015 TI - Idiopathic left ventricular tachycardia localised at the distal end of the posterior fascicle by non-contact activation mapping. PMID- 11907013 TI - Intragraft interleukin 2 mRNA expression during acute cellular rejection and left ventricular total wall thickness after heart transplantation. AB - OBJECTIVE: To assess whether diastolic graft function is influenced by intragraft interleukin 2 (IL-2) messenger RNA (mRNA) expression in rejecting cardiac allografts. DESIGN: 16 recipients of cardiac allografts were monitored during the first three months after transplantation. The presence of IL-2 mRNA in endomyocardial biopsies (n = 123) was measured by reverse transcriptase polymerase chain reaction. To determine heart function, concurrent M mode and two dimensional Doppler echocardiograms were analysed. RESULTS: Histological signs of acute rejection (International Society for Heart and Lung Transplantation (ISHLT) rejection grade > 2) were strongly associated with IL-2 mRNA expression (IL-2 mRNA was present in 12 of 20 endomyocardial biopsies (60%) with acute rejection and in 24 of 103 endomyocardial biopsies (23%) without acute rejection, p = 0.002). No significant relation was found between either histology or IL-2 mRNA expression alone and the studied echocardiographic parameters. However, stratification of the echocardiographic data into those of patients with and those without acute rejection showed that during acute rejection IL-2 mRNA expression was significantly associated with increased left ventricular total wall thickness (mean change in total wall thickness was +0.22 cm in patients with IL-2 mRNA expression versus -0.18 cm in patients without IL-2 mRNA expression, p = 0.048). CONCLUSIONS: An increase in left ventricular total wall thickness precedes IL-2 positive acute rejection after heart transplantation. Thus, cardiac allograft rejection accompanied by intragraft IL-2 mRNA expression may be indicative of more severe rejection episodes. PMID- 11907016 TI - Day-case transfer for percutaneous coronary intervention with adjunctive abciximab in acute coronary syndromes. PMID- 11907017 TI - Anaemia in chronic heart failure: what is its frequency in the UK and its underlying causes? PMID- 11907018 TI - Left ventricular diastolic function after electrical cardioversion of atrial fibrillation. PMID- 11907019 TI - Diagnostic accuracy of technician supervised and reported exercise tolerance tests. PMID- 11907020 TI - Arrhythmias in adults with congenital heart disease. PMID- 11907021 TI - Screening relatives of patients with premature coronary heart disease. PMID- 11907022 TI - The medical management of valvar heart disease. PMID- 11907023 TI - Changes in mobility account for camptothecin-induced subnuclear relocation of topoisomerase I. AB - DNA topoisomerase I is a nucleolar protein, which relocates to the nucleoplasm in response to drugs stabilizing topoisomerase I.DNA intermediates (e.g. camptothecin). Here we demonstrate that this phenomenon is solely caused by the drug's impact on the interplay between mobility and localization of topoisomerase I in a living cell nucleus. We show by photobleaching of cells expressing biofluorescent topoisomerase I-chimera that the enzyme moves continuously between nucleoli and nucleoplasm. Complex kinetics of fluorescence recovery after photobleaching indicates that two enzyme fractions with different mobility coexist in nucleoli and nucleoplasm. However, the whole complement of topoisomerase I is in continuous flux between these compartments and nucleolar accumulation can plausibly explained by the enzyme's 2-fold lesser overall mobility in nucleoli versus nucleoplasm. Upon addition of camptothecin, topoisomerase I relocates within 30 s from the nucleoli to radial nucleoplasmic structures. At these sites, the enzyme becomes retarded in a dose-dependent manner. Inside nucleoli the mobility of topoisomerase I is much less affected by camptothecin. Thus, the enzyme's distribution equilibrium is shifted toward the nucleoplasm, which causes nucleolar delocalization. In general, topoisomerase I is an entirely mobile nuclear component, unlikely to require specific signaling for movements between nuclear compartments. PMID- 11907025 TI - Structures of the human Rad17-replication factor C and checkpoint Rad 9-1-1 complexes visualized by glycerol spray/low voltage microscopy. AB - Human checkpoint Rad proteins are thought to function as damage sensors in the DNA damage checkpoint response pathway. The checkpoint proteins hRad9, hHus1, and hRad1 have limited homology to the replication processivity factor proliferating cell nuclear antigen (PCNA), and hRad17 has homology to replication factor C (RFC). Such observations have led to the proposal that these checkpoint Rad proteins may function similarly to their replication counterparts during checkpoint control. We purified two complexes formed by the checkpoint Rad proteins and investigated their structures using an electron microscopic preparative method in which the complexes are sprayed from a glycerol solution onto very thin carbon foils, decorated in vacuo with tungsten, and imaged at low voltage. We found that the hRad9, hHus1, and hRad1 proteins make a trimeric ring structure (checkpoint 9-1-1 complex) reminiscent of the PCNA ring. Similarly we found that hRad17 makes a heteropentameric complex with the four RFC small subunits (hRad17-RFC) with a deep groove or cleft and is similar to the RFC clamp loader. Therefore, our results demonstrate structural similarity between the checkpoint Rad complexes and the PCNA and RFC replication factors and thus provide further support for models proposing analogous functions for these complexes. PMID- 11907024 TI - A biotin analog inhibits acetyl-CoA carboxylase activity and adipogenesis. AB - Acetyl-CoA carboxylase catalyzes the first committed step in the synthesis of long chain fatty acids. In this study, we observed that treatment of 3T3-L1 cells with biotin chloroacetylated at the 1' nitrogen reduced the enzymatic activity of cytosolic acetyl-CoA carboxylase and concomitantly inhibited the differentiation of 3T3-L1 cells in a dose-dependent manner. Treatment with chloroacetylated biotin blocked the induction of PPARgamma, STAT1, and STAT5A expression that normally occurs with adipogenesis. Moreover, addition of chloroacetylated biotin inhibited lipid accumulation, as judged by Oil Red O staining. Our results support recent studies that indicate that acetyl-CoA carboxylase may be a suitable target for an anti-obesity therapeutic. PMID- 11907026 TI - Critical role of cAMP-response element-binding protein for angiotensin II-induced hypertrophy of vascular smooth muscle cells. AB - We reported previously an important role of cyclic AMP-response element (CRE) for the induction of interleukin-6 gene expression by angiotensin II (AngII). We examined signaling pathways that are responsible for AngII-induced phosphorylation of CRE-binding protein (CREB) at serine 133 that is a critical marker for the activation in rat vascular smooth muscle cells (VSMC). AngII time dependently induced phosphorylation of CREB with a peak at 5 min. The AngII induced phosphorylation of CREB was blocked by CV11974, an AngII type I receptor antagonist, suggesting that AngII type I receptor may mediate the phosphorylation of CREB. Inhibition of extracellular signal-regulated protein kinase (ERK) by PD98059 or inhibition of p38 mitogen-activated protein kinase (MAPK) by SB203580 partially inhibited AngII-induced CREB phosphorylation. A protein kinase A inhibitor, H89, also partially suppressed AngII-induced CREB phosphorylation. Inhibition of epidermal growth factor-receptor by AG1478 suppressed the AngII induced CREB phosphorylation as well as activation of ERK and p38MAPK. Overexpression of the dominant negative form of CREB by an adenovirus vector suppressed AngII-induced c-fos expression and incorporation of [(3)H]leucine to VSMC. These findings suggest that AngII may activate multiple signaling pathways involving two MAPK pathways and protein kinase A, all of which contribute to the activation of CREB. Transactivation of epidermal growth factor-receptor is also critical for AngII-induced CREB phosphorylation. Activation of CREB may be important for the regulation of gene expression and hypertrophy of VSMC induced by AngII. PMID- 11907027 TI - Regulation of proto-Dbl by intracellular membrane targeting and protein stability. AB - The pleckstrin homology (PH) domain of onco-Dbl, a guanine nucleotide exchange factor (GEF) for Cdc42 and RhoA GTPases, interacts with phosphoinositides (PIPs). This interaction modulates both the GEF activity and the targeting to the plasma membrane of onco-Dbl. Conversely, we have previously shown that in proto-Dbl an intramolecular interaction between the N-terminal domain and the PH domain imposes a negative regulation on both the DH and PH functions, suppressing its transforming activity. Here we have further investigated the mode of regulation of proto-Dbl by generating proto-Dbl mutants deleted of the last C-terminal 50 amino acids, which contain a PEST motif, and/or unable to bind to PIPs due to substitutions of the positively charged residues of the PH domain. The PH mutants of proto-Dbl retained a relative weak GEF activity toward Cdc42 and RhoA in vitro, but their RhoA activating potential was impaired in vivo. Further, these mutants lost both the plasma membrane targeting and the transforming activities, contrary to the PH mutants of onco-Dbl that retained the exchange activity both in vitro and in vivo and showed significant, but partially, reduced transforming activity. Deletion of the C-terminal sequences from onco-Dbl did not affect its function, whereas similar deletion of proto-Dbl led to an increase of transforming activity. Analysis of the half-life of the proto-Dbl mutants revealed that deletion of the C-terminal sequences increases the stability of the protein. Overall, the transformation potential of proto-Dbl mutants was associated with an augmented localization of the protein to the plasma membrane and a strong activation of Jun N-terminal kinase activity and transcription of cyclin D1. Together with previous observations, these data suggest that the biological activity of proto-Dbl is tightly regulated by a combination of mechanisms that involve intramolecular interaction, PH binding to PIPs, and the N and C-terminal domain-dependent turnover of the protein. PMID- 11907028 TI - Src-mediated inter-receptor cross-talk between the Na+/K+-ATPase and the epidermal growth factor receptor relays the signal from ouabain to mitogen activated protein kinases. AB - Binding of ouabain to Na(+)/K(+)-ATPase activates tyrosine phosphorylation of the epidermal growth factor receptor (EGFR), Src, and p42/44 mitogen-activated protein kinases (MAPKs) in both cardiac myocytes and A7r5 cells. Here, we explored the roles of Src and the EGFR in the ouabain-invoked pathways that lead to the activation of MAPKs. Exposure of A7r5 and LLC-PK1 cells to ouabain caused a dose-dependent inhibition of Na(+)/K(+)-ATPase activity, which correlated well with ouabain-induced activation of Src and MAPKs in these cells. Immunoprecipitation experiments showed that ouabain stimulated Src binding to Na(+)/K(+)-ATPase in a dose- and time-dependent manner and increased phosphorylation of Src at Tyr(418) but had no effect on Tyr(529) phosphorylation. Ouabain failed to activate MAPKs in A7r5 cells that were pretreated with the Src inhibitor PP2 and in SYF cells in which Src family kinases are knocked out. Preincubation with AG1478, but not AG1295, also blocked the effects of ouabain on p42/44 MAPKs in A7r5 cells. Significantly, both herbimycin A and PP2 abrogated ouabain-induced but not epidermal growth factor-induced Src binding to the EGFR and the subsequent EGFR tyrosine phosphorylation. Ouabain also failed to affect tyrosine phosphorylation of the EGFR in SYF cells. In addition, unlike epidermal growth factor, ouabain did not increase EGFR autophosphorylation at Tyr(1173). These findings clearly indicate that ouabain transactivates the EGFR by activation of Src and stimulation of Src binding to the EGFR. Furthermore, we found that the transactivated EGFR was capable of recruiting and phosphorylating the adaptor protein Shc. This resulted in increased binding of another adaptor protein Grb2 to the Src-EGFR complex and the subsequent activation of Ras and MAPKs. Taken together, these new findings suggest that Src mediates the inter receptor cross-talk between Na(+)/K(+)-ATPase and the EGFR to transduce the signals from ouabain to the Ras/MAPK cascade. PMID- 11907029 TI - Sterol regulatory element-binding protein-1 interacts with the nuclear thyroid hormone receptor to enhance acetyl-CoA carboxylase-alpha transcription in hepatocytes. AB - In previous work, we characterized a 3,5,3'-triiodothyronine response element (T3RE) in acetyl-CoA carboxylase-alpha (ACCalpha) promoter 2 that mediated 3,5,3' triiodothyronine (T3) regulation of ACCalpha transcription in chick embryo hepatocytes. Sequence comparison analysis revealed the presence of sterol regulatory element-1 (SRE-1) located 5 bp downstream of the ACCalpha T3RE. Here, we investigated the role of this SRE-1 in modulating T3 regulation of ACCalpha transcription. Transfection analyses demonstrated that the SRE-1 enhanced T3 induced ACCalpha transcription by more than 2-fold in hepatocytes. The effect of the SRE-1 on T3 responsiveness required the presence of the T3RE in its native orientation. In pull-down experiments, the mature form of sterol regulatory element-binding protein-1 (SREBP-1) specifically bound the alpha-isoform of the nuclear T3 receptor (TR), and the presence of T3 enhanced this interaction. A region of TRalpha containing the DNA-binding domain plus flanking sequences (amino acids 21-157) was required for interaction with SREBP-1, and a region of SREBP-1 containing the basic helix-loop-helix-leucine zipper domain (amino acids 300-389) was required for interaction with TRalpha. In gel mobility shift experiments, TRalpha, retinoid X receptor-alpha, and mature SREBP-1 formed a tetrameric complex on a DNA probe containing the ACCalpha T3RE and SRE-1, and the presence of T3 enhanced the formation of this complex. Formation of the tetrameric complex stabilized the binding of SREBP-1 to the SRE-1. These results indicate that SREBP-1 directly interacts with TR-retinoid X receptor in an orientation-specific manner to enhance T3-induced ACCalpha transcription in hepatocytes. T3 regulation of ACCalpha transcription in nonhepatic cell cultures such as chick embryo fibroblasts is markedly reduced compared with that of chick embryo hepatocytes. Here, we also show that alterations in SREBP expression play a role in mediating cell type-dependent differences in T3 regulation of ACCalpha transcription. PMID- 11907030 TI - Nerve growth factor specifically stimulates translation of eukaryotic elongation factor 1A-1 (eEF1A-1) mRNA by recruitment to polyribosomes in PC12 cells. AB - During postnatal brain development the level of peptide elongation factor-1A (eEF1A-1) expression declines and that of the highly homologous isoform, eEF1A-2, increases in neurons. eEF1A-1 is implicated in cytoskeletal interactions, tumorigenesis, differentiation, and the absence of eEF1A-2 is implicated in neurodegeneration in the mouse mutant, wasted. The translation of eEF1A-1 mRNA is up-regulated via mitogenic stimulation. However, it is not known if eEF1A-1 mRNA translation is regulated by neurotrophins or if its synthesis is differentially regulated than that of the neuronal isoform, eEF1A-2. Regulated translation of these factors by neurotrophins, particularly by the Trk class of neurotrophin receptors, would implicate them in differentiation, survival, and neuronal plasticity. In this study, we investigated the effect of nerve growth factor (NGF) stimulation on the synthesis of eEF1A-1 and eEF1A-2. We found that NGF stimulation causes a preferential synthesis of eEF1A-1 over eEF1A-2 in PC12 cells. We analyzed the co-sedimentation of eEF1A-1 mRNA with polyribosome fractions in sucrose gradients, and found that NGF stimulation enriched the presence of eEF1A-1 mRNA in polyribosomes, indicating that the translation of eEF1A-1 mRNA is regulated by NGF. Inhibitors of phosphatidylinositol 3-kinase (LY 294002), mammalian target of rapamycin (rapamycin), and the NGF receptor, TrkA (K 252a), but not of mitogen-activated protein kinase (PD 98059), prevented the recruitment of eEF1A-1 mRNA to polyribosomes. The mobilization of eEF1A-1 mRNA to polyribosomes was rapamycin-sensitive in both proliferating and differentiated PC12 cells, indicating the importance of this pathway during differentiation. Our data shows that after growth factor withdrawal, an NGF-signaling pathway stimulates eEF1A-1 mRNA translation in proliferating and differentiated PC12 cells. Therefore, eEF1A-1 mRNA is a specific translational target of TrkA signaling. PMID- 11907031 TI - Functional characterization of betaine/proline transporters in betaine accumulating mangrove. AB - Betaine is an important osmoprotectant in many plants, but its transport activity has only been demonstrated using a proline transporter from tomato, a betaine nonaccumulating plant. In this study, two full-length and one partial transporter genes were isolated from betaine-accumulating mangrove Avicennia marina. Their homologies to betaine transporters from bacteria and betaine/4-aminobutyrate transporters from mammalian cells were low but were high to proline transporters from Arabidopsis and tomato. Two full-length transporters could complement the Na(+)-sensitive phenotype of the Escherichia coli mutant deficient in betT, putPA, proP, and proU. Both transporters could efficiently take up betaine and proline with similar affinities (K(m), 0.32-0.43 mm) and maximum velocities (1.9 3.6 nmol/min/mg of protein). The uptakes of betaine and proline were significantly inhibited by mono- and dimethylglycine but only partially inhibited by betaine aldehyde, choline, and 4-aminobutyrate. Sodium and potassium chloride markedly enhanced betaine uptake rates with optimum concentrations at 0.5 m, whereas sucrose showed only modest activation. The change of amino acids Thr(290) Thr-Ser(292) in a putative periplasmic loop to Arg(290)-Gly-Arg(292) yielded the active transporter independent of salts, suggesting the positive charge induced a conformational change to the active form. These data clearly indicate that the betaine-accumulating mangrove contains betaine/proline transporters whose properties are distinct from betaine transporters of bacteria and mammalian cells. PMID- 11907032 TI - Overexpression of calreticulin modulates protein kinase B/Akt signaling to promote apoptosis during cardiac differentiation of cardiomyoblast H9c2 cells. AB - Calreticulin is a Ca(2+)-binding molecular chaperone of the lumen of the endoplasmic reticulum. Calreticulin has been shown to be essential for cardiac and neural development in mice, but the mechanism by which it functions in cell differentiation is not fully understood. To examine the role of calreticulin in cardiac differentiation, the calreticulin gene was introduced into rat cardiomyoblast H9c2 cells, and the effect of calreticulin overexpression on cardiac differentiation was examined. Upon culture in a differentiation medium containing fetal calf serum (1%) and retinoic acid (10 nm), cells transfected with the calreticulin gene were highly susceptible to apoptosis compared with controls. In the gene-transfected cells, protein kinase B/Akt signaling was significantly suppressed during differentiation. Furthermore, protein phosphatase 2A, a Ser/Thr protein phosphatase, was significantly up-regulated, implying suppression of Akt signaling due to dephosphorylation of Akt by the up-regulated protein phosphatase 2A via regulation of Ca(2+) homeostasis. Thus, overexpression of calreticulin promotes differentiation-dependent apoptosis in H9c2 cells by suppressing the Akt signaling pathway. These findings indicate a novel mechanism by which cytoplasmic Akt signaling is modulated to cause apoptosis by a resident protein of the endoplasmic reticulum, calreticulin. PMID- 11907033 TI - Identification of a novel Na+-independent acidic amino acid transporter with structural similarity to the member of a heterodimeric amino acid transporter family associated with unknown heavy chains. AB - We identified a novel Na(+)-independent acidic amino acid transporter designated AGT1 (aspartate/glutamate transporter 1). AGT1 exhibits the highest sequence similarity (48% identity) to the Na(+)-independent small neutral amino acid transporter Asc (asc-type amino acid transporter)-2 a member of the heterodimeric amino acid transporter family presumed to be associated with unknown heavy chains (Chairoungdua, A., Kanai, Y., Matsuo, H., Inatomi, J., Kim, D. K., and Endou, H. (2001) J. Biol. Chem. 276, 49390-49399). The cysteine residue responsible for the disulfide bond formation between transporters (light chains) and heavy chain subunits of the heterodimeric amino acid transporter family is conserved for AGT1. Because AGT1 solely expressed or coexpressed with already known heavy chain 4F2hc (4F2 heavy chain) or rBAT (related to b(0,+)-amino acid transporter) did not induce functional activity, we generated fusion proteins in which AGT1 was connected with 4F2hc or rBAT. The fusion proteins were sorted to the plasma membrane and expressed the Na(+)-independent transport activity for acidic amino acids. Distinct from the Na(+)-independent cystine/glutamate transporter xCT structurally related to AGT1, AGT1 did not accept cystine, homocysteate, and l alpha-aminoadipate and exhibited high affinity to aspartate as well as glutamate, suggesting that the negative charge recognition site in the side chain-binding site of AGT1 would be closer to the alpha-carbon binding site compared with that of xCT. The AGT1 message was predominantly expressed in kidney. In mouse kidney, AGT1 protein was present in the basolateral membrane of the proximal straight tubules and distal convoluted tubules. In the Western blot analysis, AGT1 was detected as a high molecular mass band in the nonreducing condition, whereas the band shifted to a 40-kDa band corresponding to the AGT1 monomer in the reducing condition, suggesting the association of AGT1 with other protein via a disulfide bond. The finding of AGT1 and Asc-2 has established a new subgroup of the heterodimeric amino acid transporter family whose members associate not with 4F2hc or rBAT but with other unknown heavy chains. PMID- 11907034 TI - Structure determination of T cell protein-tyrosine phosphatase. AB - Protein-tyrosine phosphatase 1B (PTP1B) has recently received much attention as a potential drug target in type 2 diabetes. This has in particular been spurred by the finding that PTP1B knockout mice show increased insulin sensitivity and resistance to diet-induced obesity. Surprisingly, the highly homologous T cell protein-tyrosine phosphatase (TC-PTP) has received much less attention, and no x ray structure has been provided. We have previously co-crystallized PTP1B with a number of low molecular weight inhibitors that inhibit TC-PTP with similar efficiency. Unexpectedly, we were not able to co-crystallize TC-PTP with the same set of inhibitors. This seems to be due to a multimerization process where residues 130-132, the DDQ loop, from one molecule is inserted into the active site of the neighboring molecule, resulting in a continuous string of interacting TC-PTP molecules. Importantly, despite the high degree of functional and structural similarity between TC-PTP and PTP1B, we have been able to identify areas close to the active site that might be addressed to develop selective inhibitors of each enzyme. PMID- 11907035 TI - Parallel activation of phosphatidylinositol 4-kinase and phospholipase C by the extracellular calcium-sensing receptor. AB - The calcium-sensing receptor (CaR) is a G protein-coupled receptor that regulates physiological processes including Ca(2+) metabolism, Na(+), Cl(-), K(+), and H(2)0 balance, and the growth of some epithelial cells through diverse signaling pathways. Although many effects of CaR are mediated by the heterotrimeric G proteins Galpha(q) and Galpha(i), not all signaling pathways regulated by CaR have been identified. We used human embryonic kidney (HEK)-293 cells that stably express human CaR to study the regulation of inositol lipid metabolism by CaR. The nonfunctional mutant CaR(R796W) was used as a negative control. We found that CaR regulates phosphatidylinositol (PI) 4-kinase, the first step in inositol lipid biosynthesis. In cells pretreated with to inhibit phospholipase C activation and to block the degradation of PI 4,5-bisphosphate to form [(3)H]inositol trisphosphate (IP(3)), CaR stimulated the accumulation of [(3)H]PI monophosphate (PIP). Additionally, wortmannin, an inhibitor of both PI 3-kinase and type III PI 4-kinase, blocked CaR-stimulated accumulation of [(3)H]PIP and inhibited [(3)H]IP(3) production. CaR-stimulated inositol lipid synthesis was attributable to PI 4-kinase and not PI 3-kinase because CaR did not activate Akt, a downstream target of PI 3-kinase. CaR associates with PI 4-kinase based on the findings that CaR and the 110-kDa PI 4-kinase beta can be co-immunoprecipitated with antibodies against either CaR or PI 4-kinase. The PI-4 kinase in co immunoprecipitates with anti-CaR antibody was activated in Ca(2+)-stimulated HEK 293 cells, which stably express the wild type CaR. Pertussis toxin did not affect the formation of [(3)H]IP(3) or the rise in intracellular Ca(2+) (Handlogten, M. E., Huang, C. F., Shiraishi, N., Awata, H., and Miller, R. T. (2001) J. Biol. Chem. 276, 13941-13948). RGS4, an accelerator of GTPase activity of members of the Galpha(i) and Galpha(q) families, attenuated the CaR-stimulated PLC activation and IP(3) accumulation, which is mediated by Galpha(q), but did not inhibit CaR-stimulated [(3)H]PIP formation. In HEK-293 cells, which express wild type CaR, Rho was enriched in immune complexes co-immunoprecipitated with the anti-CaR antibody. C(3) toxin, an inhibitor of Rho, also inhibited the CaR stimulated [(3)H]IP(3) production but did not lead to CaR-stimulated [(3)H]PIP formation, reflecting inhibition of PI 4-kinase. Taken together, our data demonstrate that CaR stimulates PI 4-kinase, the first step in inositol lipid biosynthesis conversion of PI to PI 4-P by Rho-dependent and Galpha(q)- and Galpha(i)-independent pathways. PMID- 11907036 TI - Dimerization and release of molecular chaperone inhibition facilitate activation of eukaryotic initiation factor-2 kinase in response to endoplasmic reticulum stress. AB - Phosphorylation of eukaryotic initiation factor-2 (eIF2) by pancreatic eIF2 kinase (PEK), induces a program of translational expression in response to accumulation of malfolded protein in the endoplasmic reticulum (ER). This study addresses the mechanisms activating PEK, also designated PERK or EIF2AK3. We describe the characterization of two regions in the ER luminal portion of the transmembrane PEK that carry out distinct functions in the regulation of this eIF2 kinase. The first region mediates oligomerization between PEK polypeptides, and deletion of this portion of PEK blocked induction of eIF2 kinase activity. The second characterized region of PEK facilitates interaction with ER chaperones. In the absence of stress, PEK associates with ER chaperones GRP78 (BiP) and GRP94, and this binding is released in response to ER stress. ER luminal sequences flanking the transmembrane domain are required for GRP78 interaction, and deletion of this portion of PEK led to its activation even in the absence of ER stress. These results suggest that this ER chaperone serves as a repressor of PEK activity, and release of ER chaperones from PEK when misfolded proteins accumulate in the ER induces gene expression required to enhance the protein folding capacity of the ER. PMID- 11907037 TI - Characterization and expression of L-amino acid oxidase of mouse milk. AB - l-Amino acid oxidase (LAO) was purified from mouse milk. LAO reacted with l-amino acids in an apparent order of Phe > Met, Tyr > Cys, Leu > His other 11 amino acids tested and produced H(2)O(2) in a dose- and time-dependent manner. LAO in milk had a molecular mass of about 113 kDa and was converted to a 60-kDa protein by SDS-PAGE. LAO consisted of two subunits. The N- and C-terminal amino acid sequence determination followed by cDNA cloning showed that the 60-kDa protein consisted of 497 amino acids. LAO mRNA spanned about 2.0 kb, and its expression was found only in the mammary epithelial cells. Glucocorticoid was essential for LAO gene expression. Thus, the LAO gene is expressed acutely upon the onset of milk synthesis. LAO mRNA increased 1 day before parturition, peaked during early to mid-lactation, and decreased at the end of lactation. This is the first demonstration showing that LAO is present in milk. Mastitis is caused by an intramammary bacterial infection. As mouse milk produced H(2)O(2) using endogenous free amino acids, we suggest that LAO, together with free amino acids, is responsible for killing bacteria in the mammary gland. PMID- 11907038 TI - Telomeric localization of the vertebrate-type hexamer repeat, (TTAGGG)n, in the wedgeshell clam Donax trunculus and other marine invertebrate genomes. AB - The hexamer repeat sequence (TTAGGG)(n), found at the ends of all vertebrate chromosomes, was previously identified as the main building element of one member of a HindIII satellite DNA family characterized in the genome of the bivalve mollusc Donax trunculus. It was also found in 22 perfect tandem repeats in a cloned junction region juxtaposed to the proper satellite sequence, from which the DNA tract encompassing the clustered tandem copies was excised and subcloned. Here, the chromosomal distribution of (TTAGGG)(n) sequences in the Donax was studied by the sensitivity to Bal31 exonuclease digestion, fluorescence in situ hybridization (FISH) on metaphase chromosomes and rotating-field gel electrophoresis. To verify the occurrence of the hexamer repeat in the genomes of taxonomically related molluscs and other marine invertebrates, genomic DNA from the mussel Mytilus galloprovincialis and the echinoderm Holothuria tubulosa was also analyzed. The kinetics of Bal31 hydrolysis of high molecular mass DNA from the three marine invertebrates revealed a marked decrease over time of the hybridization with the cloned (TTAGGG)(22) sequence, concomitantly with a progressive shortening of the positively reacting DNA fragments. This revealed a marked susceptibility to exonuclease consistent with terminal positioning on the respective chromosomal DNAs. In full agreement, FISH results with the (TTAGGG)(22) probe showed that the repeat appears located in telomeric regions in all chromosomes of both bivalve molluscs. The presence of (TTAGGG)(n) repeat tracts in marine invertebrate telomeres points to its wider distribution among eukaryotic organisms and suggests an ancestry older than originally presumed from its vertebrate distinctiveness. PMID- 11907039 TI - Presteady-state analysis of a single catalytic turnover by Escherichia coli uracil-DNA glycosylase reveals a "pinch-pull-push" mechanism. AB - Uracil-DNA glycosylase catalyzes the excision of uracils from DNA via a mechanism where the uracil is extrahelically flipped out of the DNA helix into the enzyme active site. A conserved leucine is inserted into the DNA duplex space vacated by the uracil leading to the paradigmatic "push-pull" mechanism of nucleotide flipping. However, the order of these two steps during catalysis has not been conclusively established. We report a complete kinetic analysis of a single catalytic turnover using a hydrolyzable duplex oligodeoxyribonucleotide substrate containing a uracil:2-aminopurine base pair. Rapid chemical-quenched-flow methods defined the kinetics of excision at the active site during catalysis. Stopped flow fluorometry monitoring the 2-aminopurine fluorescence defined the kinetics of uracil flipping. Parallel experiments detecting the protein fluorescence showed a slower Leu(191) insertion step occurring after nucleotide flipping but before excision. The inserted Leu(191) acts as a doorstop to prevent the return of the flipped-out uracil residue, thereby facilitating the capture of the uracil in the active site and does not play a direct role in "pushing" the uracil out of the DNA helix. The results define for the first time the proper sequence of events during a catalytic cycle and establish a "pull-push", as opposed to a "push-pull", mechanism for nucleotide flipping. PMID- 11907040 TI - High resolution X-ray structure of galactose mutarotase from Lactococcus lactis. AB - Galactose mutarotase plays a key role in normal galactose metabolism by catalyzing the interconversion of beta-D-galactose and alpha-D-galactose. Here we describe the three-dimensional architecture of galactose mutarotase from Lactococcus lactis determined to 1.9-A resolution. Each subunit of the dimeric enzyme displays a distinctive beta-sandwich motif. This tertiary structural element was first identified in beta-galactosidase and subsequently observed in copper amine oxidase, hyaluronate lyase, chondroitinase, and maltose phosphorylase. Two cis-peptides are found in each subunit, namely Pro(67) and Lys(136). The active site is positioned in a rather open cleft, and the electron density corresponding to the bound galactose unequivocally demonstrates that both anomers of the substrate are present in the crystalline enzyme. Those residues responsible for anchoring the sugar to the protein include Arg(71), His(96), His(170), Asp(243), and Glu(304). Both His(96) and His(170) are strictly conserved among mutarotase amino acid sequences determined thus far. The imidazole nitrogens of these residues are located within hydrogen bonding distance to the C-5 oxygen of galactose. Strikingly, the carboxylate group of Glu(304) is situated at approximately 2.7 A from the 1'-hydroxyl group of galactose, thereby suggesting its possible role as a general acid/base group. PMID- 11907041 TI - Biochemical and structural definition of the l-afadin- and actin-binding sites of alpha-catenin. AB - alpha-Catenin is an integral component of adherens junctions, where it links cadherins to the actin cytoskeleton. alpha-Catenin is also required for the colocalization of the nectin/afadin/ponsin adhesion system to adherens junctions, and it specifically associates with the nectin-binding protein afadin. A proteolytic fragment of alpha-catenin, residues 385-651, contains the afadin binding site. The three-dimensional structure of this fragment comprises two side by-side four-helix bundles, both of which are required for afadin binding. The alpha-catenin fragment 385-651 binds afadin more strongly than the full-length protein, suggesting that the full-length protein harbors a cryptic binding site for afadin. Comparison of the alpha-catenin 385-651 structure with the recently solved structure of the alpha-catenin M-fragment (Yang, J., Dokurno, P., Tonks, N. K., and Barford, D. (2001) EMBO J. 20, 3645-3656) reveals a surprising flexibility in the orientation of the two four-helix bundles. alpha-Catenin and the actin-binding protein vinculin share sequence and most likely structural similarity within their actin-binding domains. Despite this homology, actin binding requires additional sequences adjacent to this region. PMID- 11907042 TI - Biosynthesis of surfactant protein C (SP-C). Sorting of SP-C proprotein involves homomeric association via a signal anchor domain. AB - Rat surfactant protein C (SP-C) is synthesized as a 194-amino acid propeptide (SP C-(1-194)) that is directed to the distal secretory pathway and proteolytically processed as an integral membrane protein to yield its mature form. We had shown previously that trafficking of proSP-C is mediated both by a signal anchor domain contained within the mature SP-C sequence and by a targeting domain in the NH(2) flanking propeptide. Based on evidence from other integral membrane proteins, we hypothesized that proSP-C targeting is effected by oligomerization of proSP-C monomers. To evaluate this in vitro, cDNA constructs encoding for either wild type proSP-C (pcDNA3/SP-C-(1-194)) or heterologous fusion proteins containing green fluorescent protein (EGFP) linked to SP-C-(1-194) (EGFP/SP-C-(1-194)), to mutant proSP-C lacking the NH(2) targeting domain (EGFP/SP-C-(24-194)), or to mature SP-C alone (EGFP/SP-C-(24-58)) were produced. In transfected A549 cells, fluorescence microscopy revealed that pcDNA3/SP-C-(1-194) and EGFP/SP-C-(1-194) were each expressed in CD63 (+), EEA1 (-) cytoplasmic vesicles. Expression of EGFP/SP-C-(24-194) or EGFP/SP-C-(24-58) resulted in translocation but retention in early compartments. When co-transfected with pcDNA3/SP-C-(1-194), both EGFP/SP C-(24-194) and EGFP/SP-C-(24-58) were directed to CD63 (+) vesicles that also contained SP-C-(1-194). In contrast, trafficking of a folding mutant that forms juxtanuclear aggregates, EGFP/SP-C(C122/186G), was not corrected by cotransfection with pcDNA3/SP-C-(1-194). Chemical cross-linking studies of transfected cell lysates with bismaleimidohexane produced multimeric forms of both EGFP/SP-C-(1-194) and EGFP/SP-C-(24-58). These results indicate that sorting involves oligomeric association of proSP-C monomers mediated by the mature SP-C domain. Heteromeric assembly allows wild type proSP-C to facilitate trafficking of SP-C mutants with intact transmembrane domains but lacking targeting signals. We speculate that heterotypic oligomerization of wild type with SP-C folding mutants produces a dominant negative thus contributing to the pathology of chronic lung disease associated with patients heterozygous for mutant SP-C alleles. PMID- 11907043 TI - The role of mitochondrial porins and the permeability transition pore in learning and synaptic plasticity. AB - Mitochondrial outer membrane permeability is conferred by a family of porin proteins. Mitochondrial porins conduct small molecules and constitute one component of the permeability transition pore that opens in response to apoptotic signals. Because mitochondrial porins have significant roles in diverse cellular processes including regulation of mitochondrial ATP and calcium flux, we sought to determine their importance in learning and synaptic plasticity in mice. We show that fear conditioning and spatial learning are disrupted in porin-deficient mice. Electrophysiological recordings of porin-deficient hippocampal slices reveal deficits in long and short term synaptic plasticity. Inhibition of the mitochondrial permeability transition pore by cyclosporin A in wild-type hippocampal slices reproduces the electrophysiological phenotype of porin deficient mice. These results demonstrate a dynamic functional role for mitochondrial porins and the permeability transition pore in learning and synaptic plasticity. PMID- 11907044 TI - Proteolytic processing of low density lipoprotein receptor-related protein mediates regulated release of its intracellular domain. AB - The low density lipoprotein (LDL) receptor-related protein (LRP) is a multifunctional cell surface receptor that interacts through its cytoplasmic tail with adaptor and scaffold proteins that participate in cellular signaling. Its extracellular domain, like that of the signaling receptor Notch and of amyloid precursor protein (APP), is proteolytically processed at multiple positions. This similarity led us to investigate whether LRP, like APP and Notch, might also be cleaved at a third, intramembranous or cytoplasmic site, resulting in the release of its intracellular domain. Using independent experimental approaches we demonstrate that the cytoplasmic domain is released by a gamma-secretase-like activity and that this event is modulated by protein kinase C. Furthermore, cytoplasmic adaptor proteins that bind to the LRP tail affect the subcellular localization of the free intracellular domain and may regulate putative signaling functions. Finally, we show that the degradation of the free tail fragment is mediated by the proteasome. These findings suggest a novel role for the intracellular domain of LRP that may involve the subcellular translocation of preassembled signaling complexes from the plasma membrane. PMID- 11907045 TI - L-selectin dimerization enhances tether formation to properly spaced ligand. AB - Selectin counterreceptors are glycoprotein scaffolds bearing multiple carbohydrate ligands with exceptional ability to tether flowing cells under disruptive shear forces. Bond clusters may facilitate formation and stabilization of selectin tethers. L-selectin ligation has been shown to enhance L-selectin rolling on endothelial surfaces. We now report that monoclonal antibodies-induced L-selectin dimerization enhances L-selectin leukocyte tethering to purified physiological L-selectin ligands and glycopeptides. Microkinetic analysis of individual tethers suggests that leukocyte rolling is enhanced through the dimerization-induced increase in tether formation, rather than by tether stabilization. Notably, L-selectin dimerization failed to augment L-selectin mediated adhesion below a threshold ligand density, suggesting that L-selectin dimerization enhanced adhesiveness only to properly clustered ligand. In contrast, an epidermal growth factor domain substitution of L-selectin enhanced tether formation to L-selectin ligands irrespective of ligand density, suggesting that this domain controls intrinsic ligand binding properties of L-selectin without inducing L-selectin dimerization. Strikingly, at low ligand densities, where L-selectin tethering was not responsive to dimerization, elevated shear stress restored sensitivity of tethering to selectin dimerization. This is the first indication that shear stress augments effective selectin ligand density at local contact sites by promoting L-selectin encounter of immobilized ligand. PMID- 11907047 TI - Evolution of voltage-gated Na(+) channels. AB - Voltage-gated Na(+) channels play important functional roles in the generation of electrical excitability in most vertebrate and invertebrate species. These channels are members of a superfamily that includes voltage-gated K(+), voltage gated Ca(2+) and cyclic-nucleotide-gated channels. There are nine genes encoding voltage-gated Na(+) channels in mammals, with a tenth homologous gene that has not been shown to encode a functional channel. Other vertebrate and invertebrate species have a smaller number of Na(+) channel genes. The mammalian genes can be classified into five branches in a phylogenetic tree, and they are localized on four chromosomes. Four of the branches representing the four chromosomal locations probably resulted from the chromosomal duplications that led to the four Hox gene clusters. These duplications occurred close to the emergence of the first vertebrates. The fifth branch probably evolved from a separate ancestral Na(+) channel gene. There are two branches in the invertebrate tree, although members of only one of those branches have been demonstrated to encode functional voltage-gated Na(+) channels. It is possible that the other branch may have diverged, so that its members do not represent true voltage-gated Na(+) channels. Vertebrate and invertebrate Na(+) channels appear to be derived from a single primordial channel that subsequently evolved independently in the two lineages. PMID- 11907048 TI - The visual centring response in desert ants, Cataglyphis fortis. AB - When negotiating their way through cluttered environments, desert ants, Cataglyphis fortis, tend to run along the midlines of the alleys formed by adjacent low shrubs. This 'centring response' was investigated by inducing foraging ants to walk through artificial channels. The sidewalls of the channel were either homogeneously black or provided with stationary or moving black-and white gratings. The speed of motion and the spatial period of the gratings and the height of the walls could be varied independently on the left-hand and right hand sides of the channel. The results clearly show that the ants, while exhibiting their centring responses, try to balance neither the self-induced image speeds nor the contrast frequencies seen in their left and right visual fields, but the vertical angle subtended by the landmarks on either side. When manoeuvring through the channel, the ants always adjust the lateral positions of their walking trajectories in such a way that the vertical angles subtended by the walls are identical for both eyes. PMID- 11907049 TI - Carbonic anhydrase in the midgut of larval Aedes aegypti: cloning, localization and inhibition. AB - The larval mosquito midgut exhibits one of the highest pH values known in a biological system. While the pH inside the posterior midgut and gastric caeca ranges between 7.0 and 8.0, the pH inside the anterior midgut is close to 11.0. Alkalization is likely to involve bicarbonate/carbonate ions. These ions are produced in vivo by the enzymatic action of carbonic anhydrase. The purpose of this study was to investigate the role of this enzyme in the alkalization mechanism, to establish its presence and localization in the midgut of larval Aedes aegypti and to clone and characterize its cDNA. Here, we report the physiological demonstration of the involvement of carbonic anhydrase in midgut alkalization. Histochemistry and in situ hybridization showed that the enzyme appears to be localized throughout the midgut, although preferentially in the gastric caeca and posterior regions with specific cellular heterogeneity. Furthermore, we report the cloning and localization of the first carbonic anhydrase from mosquito larval midgut. A cDNA clone from Aedes aegypti larval midgut revealed sequence homology to alpha-carbonic anhydrases from vertebrates. Bioinformatics indicates the presence of at least six carbonic anhydrases or closely related genes in the genome of another dipteran, the fruit fly Drosophila melanogaster. Molecular analyses suggest that the larval mosquito may also possess multiple forms. PMID- 11907050 TI - How does a fur seal mother recognize the voice of her pup? An experimental study of Arctocephalus tropicalis. AB - In the subantarctic fur seal Arctocephalus tropicalis, mothers leave their pups during the rearing period to make long and frequent feeding trips to sea. When a female returns from the ocean, she has to find her pup among several hundred others. Taking into account both spectral and temporal domains, we investigated the individual vocal signature occurring in the 'female attraction call' used by pups to attract their mother. We calculated the intra- and inter-individual variability for each measured acoustic cue to isolate those likely to contain information about individual identity. We then tested these cues in playback experiments. Our results show that a female pays particular attention to the lower part of the signal spectrum, the fundamental frequency accompanied by its first two harmonics being sufficient to elicit reliable recognition. The spectral energy distribution is also important for the recognition process. Of the temporal features, frequency modulation appears to be a key component for individual recognition, whereas amplitude modulation is not implicated in the identification of the pup's voice by its mother. We discuss these results with respect to the constraints imposed on fur seals by a colonial way of life. PMID- 11907051 TI - Ticking of the clockwork cricket: the role of the escapement mechanism. AB - The 'clockwork cricket' model for cricket sound production suggests that the catch-and-release of the file of one forewing by the plectrum on the opposite wing act as an 'escapement' to provide the phasic impulses that initiate and sustain the vibration of the resonant regions of the wings from which the sounds are produced. The action of the escapement produces the familiar ticking sound of clocks. The higher-frequency components of the songs of twelve species of cricket were analysed after removing the dominant low-frequency components and amplifying the remaining higher-frequency components. In normal song pulses of all species, the higher-frequency components showed a close phase-locking to the waveform of the dominant frequency, but the amplitude of the higher-frequency components did not correlate with that at the dominant frequency. Anomalous pulses occurred spontaneously in the songs of several species: multimodal, interrupted or curtailed pulses are described. In all of these, the anomalous pulse envelope was associated with changes in the amplitude and/or instantaneous frequency of the higher-frequency components of the sound. A model of the escapement suggests that the frequency of the residual components of the song depends on the symmetry of action of the plectrum on the teeth of the file. PMID- 11907052 TI - Kinematics and hydrodynamics of an invertebrate undulatory swimmer: the damsel fly larva. AB - The kinematics and hydrodynamics of free-swimming larvae of Enallagma cyathigerum were investigated using videography combined with a simple wake visualisation technique (tracer dyes). Damsel-fly larvae are undulatory swimmers with two distinct styles of movement: 'slow' swimming, in which body undulation is assisted by paddling of the legs, and 'fast' swimming, in which the legs are inactive. In both cases, the wake consists of discrete ring vortices shed from the caudal fin at the end of each half-stroke. The vortices propagate away from the mid-line, alternately to one side of the body then the other, at an angle of 67 degrees from dead aft. There is no aft-flowing jet such as that observed in the wakes of continuously swimming fish that use caudal fin propulsion. The estimated momentum within the vortices, and the resultant thrust on the body are in tolerable agreement with calculations based on the large-amplitude bulk momentum model of fish locomotion. However, the drag on the body is not known, so it cannot be concluded with certainty that a force balance exists. The agreement between experiment and prediction gives confidence to the idea that most, if not all, of the vorticity generated by the swimming larva is located within the observable wake elements. PMID- 11907053 TI - Evolutionary determinants of normal arterial plasma pH in ectothermic vertebrates. AB - Mean values of normal arterial pH in different species of fish, amphibians and reptiles at 15 and 25 degrees C, taken from the literature, are negatively correlated with arterial P(CO(2)) and plasma [Na(+)]. At either temperature, the data accord with the hypothesis that extracellular acid-base homeostasis evolved to maintain an optimal pH at particular cell-surface sites that are similar in all species. These hypothetical sites bear fixed negative charges that attract H(+), but which are partially screened by Na(+); for the surface pH to be constant, the bulk interstitial pH should then vary inversely with [Na(+)], as is the case. At the same time, the bulk interstitial fluid must be more acid than arterial plasma by an amount that increases with decreasing arterial P(CO(2)). With allowance made for additional screening by Ca(2+) and Mg(2+), the relevant cell-surface pH is probably approximately 6.2. PMID- 11907054 TI - Accumulation of ammonia in the body and NH(3) volatilization from alkaline regions of the body surface during ammonia loading and exposure to air in the weather loach Misgurnus anguillicaudatus. AB - The weather loach Misgurnus anguillicaudatus inhabits rice fields that experience drought in summer and ammonia loading during agricultural fertilisation. Exposure of specimens to ammonia led to the accumulation of ammonia in muscle, liver and blood. The level of ammonia reached in the plasma was the highest reported among fishes. Ammonia was not detoxified to urea, and urea excretion rate was unaffected by ammonia exposure. Fish acidified the water to reduce ammonia loading. Ammonia loading, unlike aerial exposure, did not induce glutamine synthesis, and there was no accumulation of glutamine. This is a unique observation different from those reported for other fishes in the literature. An initial switch to partial amino acid catabolism led to the accumulation of alanine and was probably associated with a decreased rate of ammonia production. Aerial exposure led to decreases in rates of ammonia and urea excretion, as well as the accumulation of tissue ammonia. As the internal ammonia levels increased, M. anguillicaudatus was able to excrete some ammonia in the gaseous form (NH(3)). The percentage of ammonia excreted as NH(3) increased with time of exposure and with increasing temperature. It appears that air-breathing through the gut is involved, with the anterior portion of the digestive tract playing a central role: it became significantly more alkaline in fish exposed to air or to environmental ammonia. The skin, which also became more alkaline during air exposure, may also be involved in ammonia volatilization in air-exposed fish. This represents the first report of a fish using volatilization of NH(3) as part of a defence against ammonia toxicity. It can be concluded that the main strategy adopted by M. anguillicaudatus confronted with ammonia loading or air exposure is to tolerate high ammonia levels in the tissues. During periods of elevated tissue ammonia levels, some ammonia is lost by volatilization via air-breathing using the gut. In addition, some ammonia may be lost across the skin during air exposure. PMID- 11907055 TI - The use of ground-borne vibrations for prey localization in the Saharan sand vipers (Cerastes). AB - Sand vipers of the genus Cerastes are specialized semi-fossorial snakes that launch predatory strikes at rodents and lizards while partially buried in the soft sand of the Saharan desert. This study attempted to document which environmental stimuli are used by these snakes as a trigger for the ambush behavior. Denervating the olfactory and vomeronasal organs produced no changes in prey capture behavior in Cerastes cerastes. Occluding the eyes of the denervated specimens resulted in significant decreases in strike distance, diversity of strike angle and strike accuracy, demonstrating the importance of visual stimuli for target acquisition in this species. Nevertheless, every olfactory-denervated, temporarily blinded specimen succeeded in capturing free-ranging mice in every trial. Presentation of chemosensory-neutral targets to the olfactory-denervated, temporarily blinded specimens resulted in similar predatory behaviors, whether the target was isothermic to the environment or heated to mammalian body temperature. Collectively, these results provide evidence for the importance of visual stimuli during foraging in C. cerastes, the first experimental evidence for foraging by vibration detection in snakes and the strongest evidence to date that snakes are capable of 'hearing' vibrational stimuli. PMID- 11907056 TI - Mechanical trade-offs explain how performance increases without increasing cost in rattlesnake tailshaker muscle. AB - Rattling by rattlesnakes is one of the fastest vertebrate movements and involves some of the highest contraction frequencies sustained by vertebrate muscle. Rattling requires higher accelerations at higher twitch frequencies, yet a previous study showed that the cost per twitch of rattling is independent of twitch frequency. We used force and video recordings over a range of temperatures to examine how western diamondback rattlesnakes (Crotalus atrox) achieve faster movements without increases in metabolic cost. The key findings are (i) that increasing muscle twitch tension trades off with decreasing twitch duration to keep the tension-time integral per twitch nearly constant over a wide range of temperatures and twitch frequencies and (ii) that decreasing lateral displacement of the rattle joint moderates the mechanical work and power required to shake the rattle at higher frequencies. These mechanical trade-offs between twitch tension and duration and between joint force and displacement explain how force, work and power increase without an increase in metabolic cost. PMID- 11907057 TI - Interspecific- and acclimation-induced variation in levels of heat-shock proteins 70 (hsp70) and 90 (hsp90) and heat-shock transcription factor-1 (HSF1) in congeneric marine snails (genus Tegula): implications for regulation of hsp gene expression. AB - In our previous studies of heat-shock protein (hsp) expression in congeneric marine gastropods of the genus Tegula, we observed interspecific and acclimation induced variation in the temperatures at which heat-shock gene expression is induced (T(on)). To investigate the factors responsible for these inter- and intraspecific differences in T(on), we tested the predictions of the 'cellular thermometer' model for the transcriptional regulation of hsp expression. According to this model, hsps not active in chaperoning unfolded proteins bind to a transcription factor, heat-shock factor-1 (HSF1), thereby reducing the levels of free HSF1 that are available to bind to the heat-shock element, a regulatory element upstream of hsp genes. Under stress, hsps bind to denatured proteins, releasing HSF1, which can now activate hsp gene transcription. Thus, elevated levels of heat-shock proteins of the 40, 70 and 90 kDa families (hsp 40, hsp70 and hsp90, respectively) would be predicted to elevate T(on). Conversely, elevated levels of HSF1 would be predicted to decrease T(on). Following laboratory acclimation to 13, 18 and 23 degrees C, we used solid-phase immunochemistry (western analysis) to quantify endogenous levels of two hsp70 isoforms (hsp74 and hsp72), hsp90 and HSF1 in the low- to mid-intertidal species Tegula funebralis and in two subtidal to low-intertidal congeners, T. brunnea and T. montereyi. We found higher endogenous levels of hsp72 (a strongly heat-induced isoform) at 13 and 18 degrees C in T. funebralis in comparison with T. brunnea and T. montereyi. However, T. funebralis also had higher levels of HSF1 than its congeners. The higher levels of HSF1 in T. funebralis cannot, within the framework of the cellular thermometer model, account for the higher T(on) observed for this species, although they may explain why T. funebralis is able to induce the heat-shock response more rapidly than T. brunnea. However, the cellular thermometer model does appear to explain the cause of the increases in T(on) that occurred during warm acclimation of the two subtidal species, in which warm acclimation was accompanied by increased levels of hsp72, hsp74 and hsp90, whereas levels of HSF1 remained stable. T. funebralis, which experiences greater heat stress than its subtidal congeners, consistently had higher ratios of hsp72 to hsp74 than its congeners, although the sum of levels of the two isoforms was similar for all three species except at the highest acclimation temperature (23 degrees C). The ratio of hsp72 to hsp74 may provide a more accurate estimate of environmental heat stress than the total concentrations of both hsp70 isoforms. PMID- 11907058 TI - Protein loss during long-distance migratory flight in passerine birds: adaptation and constraint. AB - During long-distance flights, birds catabolize not only fat but also protein. Because there is no storage form of protein, protein catabolism entails a structural or functional loss. In this study, we investigated which organs were most reduced in lean mass during different phases of fat store loss and whether protein loss can be regarded as adaptive or as a constraint. Body and organ composition were analysed both during the autumn migration over continental Europe (sample from Switzerland) and after a long-distance flight over the Sahara and the Mediterranean Sea in spring (sample from Ventotene, Italy) in four species of passerine bird: pied flycatcher Ficedula hypoleuca, willow warbler Phylloscopus trochilus, garden warbler Sylvia borin and barn swallow Hirundo rustica. Large variations in protein mass occurred when long non-stop flights were performed. After a long-distance flight, birds showed a marked increase in net protein loss when fat stores were nearing depletion (analogous to the late phase of endurance fasting when the rate of protein catabolism is increased). When fat reserves were above approximately 5-10 %, protein was derived from all organs, but particularly from the breast muscles. When fat stores diminished further and protein catabolism increased, the mass of the digestive organs was reduced fastest. When the decrease in breast muscle mass during flight was regarded in terms of potential flight performance, it appeared that the use of breast muscle protein with decreasing body mass can be regarded as adaptive as long as fat stores did not reach a critical level. Below approximately 5-10 % body fat, however, protein loss reduced flight performance. This demonstrates that the phase of fasting (the size of the remaining fat stores) is an important condition for understanding the occurrence and effects of protein loss during endurance flights. PMID- 11907059 TI - How the efficiency of rainbow trout (Oncorhynchus mykiss) ventricular muscle changes with cycle frequency. AB - Different species of animals require different cardiac performance and, in turn, their cardiac muscle exhibits different properties. A comparative approach can reveal a great deal about the mechanisms underlying myocardial contraction. Differences in myocardial Ca(2+) handling between fish and mammals suggest a greater energy cost of activation in fish. Further, while there is considerable evidence that heart rate (or cycle frequency) should have a profound effect on the efficiency of teleost cardiac muscle, this effect has been largely overlooked. We set out to determine how cycle frequency affects the power output and efficiency of rainbow trout (Oncorhynchus mykiss) ventricular muscle and to relate this to the heart's function in life. We measured power output and the rate of oxygen consumption ((O(2))) and then calculated efficiency over a physiologically realistic range of cycle frequencies. In contrast to mammalian cardiac muscle, in which (O(2)) increases with increasing heart rate, we found no significant change in (O(2)) in the teleost. However, power output increased by 25 % as cycle frequency was increased from 0.6 to 1.0 Hz, so net and total efficiency increased. A maximum total efficiency of 20 % was achieved at 0.8 Hz, whereas maximum power output occurred at 1.0 Hz. We propose that, since the heart operates continuously, high mechanical efficiency is a major adaptive advantage, particularly at lower heart rates corresponding to the more commonly used slower, sustainable swimming speeds. Efficiency was lower at the higher heart rates required during very fast swimming, which are used during escape or prey capture. If a fixed amount of Ca(2+) is released and then resequestered each time the muscle is activated, the activation cost should increase with frequency. We had anticipated that this would have a large effect on the total energy cost of contraction. However, since (O(2)) remains constant, less oxygen is consumed per cycle at high frequencies. We suggest that a constant (O(2)) would be observed if the amount of activator Ca(2+) were to decrease with frequency. This decrease in activation energy is consistent with the decrease in the systolic intracellular Ca(2+) ([Ca(2+)](i)) transient with increasing stimulation frequency seen in earlier studies. PMID- 11907061 TI - Cutting edge: polarized Th cell response induction by transferred antigen-pulsed dendritic cells is dependent on IL-4 or IL-12 production by recipient cells. AB - To assess the influence of dendritic cell (DC) production of polarizing cytokines on Th2 and Th1 development we transferred Ag-pulsed DC generated from wild-type, IL-4(-/-), or IL-12(-/-) mice into wild-type, IL-4(-/-), or IL-12(-/-) recipients. We found that DC IL-4 was not necessary for Th2 induction and that, surprisingly, DC IL-12 was not an absolute requirement for Th1 development. However, DC IL-12 production facilitated optimal Th1 response development. Critically, recipient ability to produce IL-4 or IL-12 was essential for either Th2 or Th1 development. These data help delineate the source and importance of IL 4 and IL-12 in the process of induction of polarized T cell responses by DC. PMID- 11907062 TI - Cutting edge: the minor histocompatibility antigen H60 peptide interacts with both H-2Kb and NKG2D. AB - Minor histocompatibility Ags elicit cell-mediated immune responses and graft rejection in individuals receiving MHC-matched tissues. H60 represents a dominant Ag that elicits a strong CTL response in C57BL/6 mice immunized against BALB.B. An 8-aa peptide in the H60 protein is presented by H-2K(b) and this is recognized by the TCR as an alloantigen. The intact H60 glycoprotein is a ligand for the costimulatory NKG2D receptor that is expressed by activated CD8(+) T cells. Thus, H60 may provide both an allogeneic peptide and its own costimulation. We show that mutation of an H-2K(b)-binding anchor residue in the H60 peptide completely abrogates binding of H60 glycoprotein to NKG2D and a synthetic H60 peptide partially blocks the binding of NKG2D to its ligand. Ligands of the human NKG2D receptor are remarkably polymorphic, suggesting that these may also serve as minor histocompatibility Ags. PMID- 11907063 TI - Cutting edge: susceptibility to the larval stage of the helminth parasite Taenia crassiceps is mediated by Th2 response induced via STAT6 signaling. AB - Using STAT6(-/-) BALB/c mice, we analyzed the role of STAT6-induced Th2 response in determining the outcome of murine cysticercosis caused by the helminth parasite Taenia crassiceps. After T. crassiceps infection, wild-type BALB/c mice developed a strong Th2-like response; produced high levels of IgG1, IgE, IL-4, as well as IL-13; and remained susceptible to T. crassiceps. In contrast, similarly infected STAT6(-/-) mice mounted a strong Th1-like response; produced high levels of IgG2a, IL-12, IFN-gamma, as well as nitric oxide; and efficiently controlled T. crassiceps infection. These findings demonstrate that Th2-like response induced via STAT6-mediated signaling pathway mediates susceptibility to T. crassiceps and, furthermore, that unlike the case in most helminths, immunity against T. crassiceps is mediated by a Th1-like rather than Th2-like response. PMID- 11907064 TI - Cutting edge: analysis of human V alpha 24+CD8+ NK T cells activated by alpha galactosylceramide-pulsed monocyte-derived dendritic cells. AB - Human Valpha24(+) NKT cells constitute a counterpart of mouse Valpha14(+) NKT cells, both of which use an invariant TCR-alpha chain. The human Valpha24(+) NKT cells as well as mouse Valpha14(+) NKT cells are activated by glycolipids in a CD1d-restricted manner and produce many immunomodulatory cytokines, possibly affecting the immune balance. In mice, it has been considered from extensive investigations that Valpha14(+)CD8(+) NKT cells that express invariant TCR do not exist. Here we introduce human Valpha24(+)CD8(+) NKT cells. These cells share important features of Valpha24(+) NKT cells in common, but in contrast to CD4( )CD8(-) (double-negative) or CD4(+) Valpha24(+) NKT cells, they do not produce IL 4. Our discovery may extend and deepen the research field of Valpha24(+) NKT cells as well as help to understand the mechanism of the immune balance-related diseases. PMID- 11907065 TI - Cutting edge: single-chain trimers of MHC class I molecules form stable structures that potently stimulate antigen-specific T cells and B cells. AB - We report in this work the expression and characterization of class I molecules expressed as single-chain trimers consisting of an antigenic peptide-spacer beta(2)-microglobulin-spacer H chain. Our results indicate that these single chain constructs assemble efficiently, maintain their covalent structure, and are unusually stable at the cell surface. Consequently, these constructs are at least 1000-fold less accessible to exogenous peptide than class I molecules loaded with endogenous peptides, and they are potent simulators of peptide-specific CTL and Abs. Our combined findings suggest that single-chain trimers may have applications as DNA vaccines against virus infection or tumors. PMID- 11907066 TI - Cutting edge: differential segregation of the SRC homology 2-containing protein tyrosine phosphatase-1 within the early NK cell immune synapse distinguishes noncytolytic from cytolytic interactions. AB - Inhibitory NK receptors with ligand specificity for MHC class I recruit Src homology 2-containing protein tyrosine phosphatase-1 (SHP-1) phosphatase and prevent autocytotoxicity. Activation of SHP-1 depends upon Src kinase-mediated tyrosine phosphorylation of the cytoplasmic domain of the inhibitory receptor. In this study we demonstrate, by quantitative temporal analysis, that talin, Lck, and SHP-1 are recruited to the synapse within 1 min in both cytolytic and noncytolytic conjugates. Polarization of talin and Lck rapidly disappears in the noncytolytic interactions but persists in cytolytic interactions, where protein kinase C-theta;, Src homology 2 domain-containing leukocyte protein of 76 kDa, and lysosomes are recruited within 5 min. At 1 min SHP-1 clusters in the periphery of the cytolytic synapse, whereas it clusters in the center of the noncytolytic synapse. Lck has multifocal distribution in both synapses consistent with the shared requirement for early tyrosine phosphorylation. Our studies indicate that the spatial location of SHP-1 in the synapse distinguishes noncytolytic from cytolytic interactions within the first minute. PMID- 11907067 TI - Syk regulation of phosphoinositide 3-kinase-dependent NK cell function. AB - Emerging evidence suggests that NK-activatory receptors use KARAP/DAP12, CD3zeta, and FcepsilonRIgamma adaptors that contain immunoreceptor tyrosine-based activatory motifs to mediate NK direct lysis of tumor cells via Syk tyrosine kinase. NK cells may also use DAP10 to drive natural cytotoxicity through phosphoinositide 3-kinase (PI3K). In contrast to our recently identified PI3K pathway controlling NK cytotoxicity, the signaling mechanism by which Syk associates with downstream effectors to drive NK lytic function has not been clearly defined. In NK92 cells, which express DAP12 but little DAP10/NKG2D, we now show that Syk acts upstream of PI3K, subsequently leading to the specific signaling of the PI3K-->Rac1-->PAK1-->mitogen-activated protein/extracellular signal-regulated kinase (ERK) kinase-->ERK cascade that we earlier described. Tumor cell ligation stimulated DAP12 tyrosine phosphorylation and its association with Syk in NK92 cells; Syk tyrosine phosphorylation and activation were also observed. Inhibition of Syk function by kinase-deficient Syk or piceatannol blocked target cell-induced PI3K, Rac1, PAK1, mitogen-activated protein/ERK kinase, and ERK activation, perforin movement, as well as NK cytotoxicity, indicating that Syk is upstream of all these signaling events. Confirming that Syk does not act downstream of PI3K, constitutively active PI3K reactivated all the downstream effectors as well as NK cytotoxicity suppressed in Syk-impaired NK cells. Our results are the first report documenting the instrumental role of Syk in control of PI3K-dependent natural cytotoxicity. PMID- 11907068 TI - Germinal center B cells constitute a predominant physiological source of IL-4: implication for Th2 development in vivo. AB - Protective immunity depends upon the capability of the immune system to properly adapt the response to the nature of an infectious agent. CD4(+) Th cells are implicated in this orchestration by secreting a polarized pattern of cytokines. Although Th2 development in animal models and in human cells in vitro to a large extent depends on IL-4, the nature of the cells that provide the initial IL-4 in vivo is still elusive. In this report, we describe the anatomical localization as well as the identity of IL-4-producing cells in human tonsil, a representative secondary lymphoid organ. We demonstrate that IL-4 production is a normal and intrinsic feature of germinal center (GC) B cells. We also show that expression of IL-4 is highly confined to the GCs, in which the B cells constitute the prevalent cellular source. Furthermore, immunofluorescence analysis of colon mucosa reveals a strikingly similar pattern of IL-4-expressing cells compared with tonsils, demonstrating that IL-4 production from GC B cells is not a unique feature of the upper respiratory tract. Our results show that GCs provide the most appropriate microenvironment for IL-4-dependent Th2 polarization in vivo and imply a critical role for GC B cells in this differentiation process. PMID- 11907069 TI - Transgenic expression of numb inhibits notch signaling in immature thymocytes but does not alter T cell fate specification. AB - The conserved adaptor protein Numb is an intrinsic cell fate determinant that functions by antagonizing Notch-mediated signal transduction. The Notch family of membrane receptors controls cell survival and cell fate determination in a variety of organ systems and species. Recent studies have identified a role for mammalian Notch-1 signals at multiple stages of T lymphocyte development. We have examined the role of mammalian Numb (mNumb) as a Notch regulator and cell fate determinant during T cell development. Transgenic overexpression of mNumb under the control of the Lck proximal promoter reduced expression of several Notch-1 target genes, indicating that mNumb antagonizes Notch-1 signaling in vivo. However, thymocyte development, cell cycle, and survival were unperturbed by mNumb overexpression, even though transgenic Numb was expressed at an early stage in thymocyte development (CD4(-)CD8(-)CD3(-) cells that were CD44(+)CD25(+) or CD44(-)CD25(+); double-negative 2/3). Moreover, bone marrow from mNumb transgenic mice showed no defects in thymopoiesis in competitive repopulation experiments. Our results suggest that mNumb functions as a Notch-1 antagonist in immature thymocytes, but that suppression of Notch-1 signaling at this stage does not alter gammadelta/alphabeta or CD4/CD8 T cell fate specification. PMID- 11907070 TI - Suppressors of cytokine signaling proteins are differentially expressed in Th1 and Th2 cells: implications for Th cell lineage commitment and maintenance. AB - Positive regulatory factors induced by IL-12/STAT4 and IL-4/STAT6 signaling during T cell development contribute to polarized patterns of cytokine expression manifested by differentiated Th cells. These two critical and antagonistic signaling pathways are under negative feedback regulation by a multimember family of intracellular proteins called suppressor of cytokine signaling (SOCS). However, it is not known whether these negative regulatory factors also modulate Th1/Th2 lineage commitment and maintenance. We show here that CD4(+) naive T cells constitutively express low levels of SOCS1, SOCS2, and SOCS3 mRNAs. These mRNAs and their proteins increase significantly in nonpolarized Th cells after activation by TCR signaling. We further show that differentiation into Th1 or Th2 phenotype is accompanied by preferential expression of distinct SOCS mRNA transcripts and proteins. SOCS1 expression is 5-fold higher in Th1 than in Th2 cells, whereas Th2 cells contain 23-fold higher levels of SOCS3. We also demonstrate that IL-12-induced STAT4 activation is inhibited in Th2 cells that express high levels of SOCS3 whereas IL-4/STAT6 signaling is constitutively activated in Th2 cells, but not Th1 cells, with high SOCS1 expression. These results suggest that mutually exclusive use of STAT4 and STAT6 signaling pathways by differentiated Th cells may derive in part, from SOCS3- or SOCS1-mediated repression of IL-12/STAT4- or IL-4/STAT6 signaling in Th2 and Th1 cells, respectively. Given the strong correlation between distinct patterns of SOCS expression and differentiation into the Th1 or Th2 phenotype, SOCS1 and SOCS3 proteins are therefore Th lineage markers that can serve as therapeutic targets for immune modulation therapy. PMID- 11907071 TI - Autoreactive T cells revealed in the normal repertoire: escape from negative selection and peripheral tolerance. AB - Self-reactive T cells are known to be eliminated by negative selection in the thymus or by the induction of tolerance in the periphery. However, developmental pathways that allow self-reactive T cells to inhabit the normal repertoire are not well-characterized. In this investigation, we made use of anti-small nuclear ribonucleoprotein particle (snRNP) Ig transgenic (Tg) mice (2-12 Tg) to demonstrate that autoreactive T cells can be detected and activated in both normal naive mice and autoimmune-prone MRL lpr/lpr mice. In contrast, autoreactive T cells of nonautoimmune Tg mice are tolerized by Tg B cells in the periphery. In adoptive transfer studies, autoreactive T cells from MRL lpr/lpr mice can stimulate autoantibody synthesis in nonautoimmune anti-snRNP Tg mice. Transferred CD4 T cells migrate to regions of the spleen proximal to the B cell follicles, suggesting that cognate B cell-T cell interactions are critical to the autoimmune response. Taken together, our studies suggest that anti-snRNP B cells are important APCs for T cell activation in autoimmune-prone mice. Additionally, we have demonstrated that anti-snRNP B cell anergy in nonautoimmune mice may be reversed by appropriate T cell help. PMID- 11907072 TI - IFN-gamma-inducible protein 10 (IP-10; CXCL10)-deficient mice reveal a role for IP-10 in effector T cell generation and trafficking. AB - IFN-gamma-inducible protein 10 (IP-10, CXCL10), a chemokine secreted from cells stimulated with type I and II IFNs and LPS, is a chemoattractant for activated T cells. Expression of IP-10 is seen in many Th1-type inflammatory diseases, where it is thought to play an important role in recruiting activated T cells into sites of tissue inflammation. To determine the in vivo function of IP-10, we constructed an IP-10-deficient mouse (IP-10(-/-)) by targeted gene disruption. Immunological analysis revealed that IP-10(-/-) mice had impaired T cell responses. T cell proliferation to allogeneic and antigenic stimulation and IFN gamma secretion in response to antigenic challenge were impaired in IP-10(-/-) mice. In addition, IP-10(-/-) mice exhibited an impaired contact hypersensitivity response, characterized by decreased ear swelling and reduced inflammatory cell infiltrates. T cells recovered from draining lymph nodes also had a decreased proliferative response to Ag restimulation. Furthermore, IP-10(-/-) mice infected with a neurotropic mouse hepatitis virus had an impaired ability to control viral replication in the brain. This was associated with decreased recruitment of CD4(+) and CD8(+) lymphocytes into the brain, reduced levels of IFN-gamma and the IFN-gamma-induced chemokines monokine induced by IFN-gamma (Mig, CXCL9) and IFN inducible T cell alpha chemoattractant (I-TAC, CXCL11) in the brain, decreased numbers of virus-specific IFN-gamma-secreting CD8(+) cells in the spleen, and reduced levels of demyelination in the CNS. Taken together, our data suggest a role for IP-10 in both effector T cell generation and trafficking in vivo. PMID- 11907073 TI - Donor T cell activation initiates small bowel allograft rejection through an IFN gamma-inducible protein-10-dependent mechanism. AB - The poor success in controlling small bowel (SB) allograft rejection is partially attributed to the unique immune environment in the donor intestine. We hypothesized that Ag-induced activation of donor-derived T cells contributes to the initiation of SB allograft rejection. To address the role of donor T cell activation in SB transplantation, SB grafts from DO11.10 TCR transgenic mice (BALB/c, H-2L(d+)) were transplanted into BALB/c (isografts), or single class I MHC-mismatched (L(d)-deficient) BALB/c H-2(dm2) (dm2, H-2L(d-)) mutant mice (allografts). Graft survival was followed after injection of control or antigenic OVA(323-339) peptide. Eighty percent of SB allografts developed severe rejection in mice treated with antigenic peptide, whereas <20% of allografts were rejected in mice treated with control peptide (p < 0.05). Isografts survived >30 days regardless of OVA(323-339) administration. Activation of donor T cells increased intragraft expression of proinflammatory cytokine (IFN-gamma) and CXC chemokine IFN-gamma-inducible protein-10 mRNA and enhanced activation and accumulation of host NK and T cells in SB allografts. Treatment of mice with neutralizing anti IFN-gamma-inducible protein-10 mAb increased SB allograft survival in Ag-treated mice (67%; p < 0.05) and reduced accumulation of host T cells and NK cells in the lamina propria but not mesenteric lymph nodes. These results suggest that activation of donor T cells after SB allotransplantation induces production of a Th1-like profile of cytokines and CXC chemokines that enhance infiltration of host T cells and NK cells in SB allografts. Blocking this pathway may be of therapeutic value in controlling SB allograft rejection. PMID- 11907074 TI - Interaction between Src homology 2 domain bearing protein tyrosine phosphatase substrate-1 and CD47 mediates the adhesion of human B lymphocytes to nonactivated endothelial cells. AB - CD47 modulates a variety of cell functions such as adhesion, spreading, and migration. Using a fusion protein consisting of the extracellular region of Src homology 2 domain bearing protein tyrosine phosphatase substrate-1 (SHPS-1) and the Fc portion of human Ig (SHPS-1-Ig) we investigated the effects of SHPS-1 as a ligand for CD47 on B lymphocytes. Although SHPS-1-Ig binding to human B cell lines was solely mediated via CD47, their binding capacity for soluble and immobilized SHPS-1-Ig varied among cell lines irrespective of the similar expression levels of CD47, suggesting that distinctive affinity/avidity states exist during B cell maturation. Nalm6 cell line and tonsilar B lymphocytes adhered to immobilized SHPS-1-Ig and showed polarization-like morphology. These effects of SHPS-1-Ig were blocked by anti-CD47 mAbs (B6H12 and SE5A5). Wortmannin, a phosphatidylinositol-3 kinase inhibitor, but not pertussis toxin significantly inhibited the polarization induced by the immobilized SHPS-1-Ig. Thus, SHPS-1 acts as an adhesive substrate via CD47 in human B lymphocyte. Immunohistochemical analyses indicated that SHPS-1 is expressed on high endothelial venule as well as macrophages in human tonsils. HUVECs also express SHPS-1 in the absence of any stimuli, and the adhesion of tonsilar B lymphocytes to nonactivated HUVECs was significantly inhibited by SE5A5, indicating that SHPS 1/CD47 interaction is involved in the adhesion. Our findings suggest that SHPS 1/CD47 interaction may contribute to the recruitment of B lymphocytes via endothelial cells under steady state conditions. PMID- 11907075 TI - Secondary lymphoid tissue chemokine (CCL21) activates CXCR3 to trigger a Cl- current and chemotaxis in murine microglia. AB - Microglial cells represent the major immunocompetent element of the CNS and are activated by any type of brain injury or disease. A candidate for signaling neuronal injury to microglial cells is the CC chemokine ligand CCL21, given that damaged neurons express CCL21. Investigating microglia in acute slices and in culture, we demonstrate that a local application of CCL21 for 30 s triggered a Cl(-) conductance with lasted for tens of minutes. This response was sensitive to the Cl(-) channel blockers 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid and 4-acetamide-4'-isothiocyanatostilbene, 2,2'-disulfonic acid. Moreover, CCL21 triggered a chemotaxis response, which was sensitive to Cl(-) channel blockers. In microglial cells cultured from CCR7 knockout mice, CCL21 produced the same type of Cl(-) current as well as a chemotaxis response. In contrast, in microglial cells from CXCR3 knockout mice, CCL21 triggered neither a Cl(-) conductance nor a chemotaxis response after CCL21 application. We conclude that the CCL21-induced Cl(-) current is a prerequisite for the chemotaxis response mediated by the activation of CXCR3 but not CCR7 receptors, indicating that in brain CCL21 acts via a different receptor system than in lymphoid organs. PMID- 11907076 TI - Functional consequences of noncognate interactions between CD4+ memory T lymphocytes and the endothelium. AB - The recruitment of Ag-specific T cells to sites of inflammation is a crucial step in immune surveillance. Although the molecular interactions controlling T cell extravasation are relatively well characterized, the effects of these events on T cell function are still poorly understood. Using an in vitro model of transendothelial migration of human CD4(+) memory T cells, we have investigated the molecular and functional changes induced in T cells that come into contact with the endothelium. First, we show that transendothelial migration is precluded by signals that lead to T cell division. In addition, activation of the transcription factor AP-1, without induction of NF-kappaB, is observed in T cells after noncognate interactions with endothelial cells (EC), a pattern of transcriptional regulation different from that observed in dividing T cells. Up regulation of certain adhesion (CD11a, CD49d), activation (CD69), and costimulatory (CD86) receptors accompany these transcriptional events. Most importantly, recently migrated T cells display a faster rate of migration when reseeded onto an EC monolayer. Finally, T cells become hyperresponsive to antigenic challenge after noncognate interactions with the endothelium. These effects appear not to be due to the selection of preactivated T lymphocytes, because they occur also in clonal T cell populations and appear to be mediated by alpha(L)beta(2) integrin-CD54 interactions. We conclude that CD4(+) memory T cell extravasation is accompanied by phenotypic and functional changes induced by the interactions with the EC, which favor tissue infiltration by T cells and their further activation once they reach the antigenic site. PMID- 11907077 TI - TCR activation of human T cells induces the production of exosomes bearing the TCR/CD3/zeta complex. AB - We show in this study that human T cells purified from peripheral blood, T cell clones, and Jurkat T cells release microvesicles in the culture medium. These microvesicles have a diameter of 50-100 nm, are delimited by a lipidic bilayer membrane, and bear TCR beta, CD3epsilon, and zeta. This microvesicle production is regulated because it is highly increased upon TCR activation, whereas another mitogenic signal, such as PMA and ionomycin, does not induce any release. T cell derived microvesicles also contain the tetraspan protein CD63, suggesting that they originate from endocytic compartments. They contain adhesion molecules such as CD2 and LFA-1, MHC class I and class II, and the chemokine receptor CXCR4. These transmembrane proteins are selectively sorted in microvesicles because CD28 and CD45, which are highly expressed at the plasma membrane, are not found. The presence of phosphorylated zeta in these microvesicles suggests that the CD3/TCR found in the microvesicles come from the pool of complexes that have been activated. Proteins of the transduction machinery, tyrosine kinases of the Src family, and c-Cbl are also observed in the T cell-derived microvesicles. Our data demonstrate that T lymphocytes produce, upon TCR triggering, vesicles whose morphology and phenotype are reminiscent of vesicles of endocytic origin produced by many cell types and called exosomes. Although the exact content of T cell derived exosomes remains to be determined, we suggest that the presence of TCR/CD3 at their surface makes them powerful vehicles to specifically deliver signals to cells bearing the right combination of peptide/MHC complexes. PMID- 11907078 TI - Naive CD4+ T cells exhibit distinct expression patterns of cytokines and cell surface molecules on their primary responses to varying doses of antigen. AB - The amount of an Ag used for stimulation affects the type and magnitude of T cell responses. In this study we have investigated the primary response of naive CD4(+) T cells derived from OVA-specific TCR-transgenic mice (OVA23-3) upon stimulation with varying doses of the antigenic peptide, OVA(323-339). IL-4 expression was maximal with 50 nM Ag and decreased significantly with increasing doses. In contrast, IFN-gamma expression, which was also detected at 50 nM Ag, increased with increasing doses. The expression patterns of mRNA for the Th2 specific transcription factors GATA-3 and c-Maf were parallel to that of IL-4. These expression profiles were not altered by the addition of anti-IL-4 plus anti IL-12 mAbs, suggesting that cytokine receptor signaling is not essential. Naive CD4(+) T cells stimulated with 5 nM Ag elicited IgM secretion from cocultured B cells, whereas those stimulated with 50 nM Ag or more elicited apoptosis of B cells. This may be because at lower doses of Ag (5 nM), naive CD4(+) T cells express CD40 ligand and OX40, whereas at higher doses (50 nM), they express Fas ligand. Clearly, the expression of each type of molecule depends on the Ag dose, and different molecules had different expression patterns. Thus, in the primary response, naive CD4(+) T cells can exhibit different functions depending on the dose of Ag. PMID- 11907079 TI - Phenotypic and functional differences between NKT cells colonizing splanchnic and peripheral lymph nodes. AB - NKT cells are considered unconventional T cells. First, they are restricted by a nonclassical MHC class I molecule, CD1d, which presents glycolipids; second, their TCR repertoire is very limited. After stimulation by their TCR, NKT cells rapidly release large amounts of cytokines, such as IL-4 and IFN-gamma. Little is known about NKT cells present in lymph nodes. In the present report we show that NKT cells are differently distributed in various lymph nodes and are, for instance, abundant in pancreatic and mesenteric lymph nodes of C57BL/6 mice and nonobese diabetic mice. The high frequency of NKT cells in splanchnic lymph nodes is not simply a consequence of inflammatory signals, as draining lymph nodes still contain low frequencies of NKT cells after IFA or CFA injections. NKT cells from splanchnic lymph nodes harbor a Vbeta repertoire similar to that of splenic and liver NKT cells, in contrast to peripheral NKT cells that are not biased toward Vbeta8 segments. Analysis of cytokine production by NKT cells from splanchnic lymph nodes reveals that they produce at least as much IL-4 as IFN gamma, in contrast to NKT cells from other organs (spleen, liver, and peripheral lymph nodes), which produce much more IFN-gamma than IL-4. These specific features of NKT cells from splanchnic lymph nodes might explain their protective action against the development of pathogenic Th1 cells in type 1 diabetes. PMID- 11907080 TI - Superantigen-induced TCR alpha locus secondary rearrangement: role in tolerance induction. AB - Immunization with superantigen in vivo induces transient activation of superantigen-specific T cells, followed by a superantigen-nonresponsive state. In this study, using a TCR alpha knock-in mouse in which the knock-in alpha-chain can be replaced with endogenous alpha-chain through secondary rearrangement, we show that immunization of superantigen changes the TCR alpha-chain expression on peripheral superantigen-specific T cells, induces expression of recombination activating genes, and generates DNA double-strand breaks at the TCR alpha-chain locus. These results suggest that viral superantigens are capable of inducing peripheral TCR revision. Our findings thus provide a new perspective on pathogen immune system interaction. PMID- 11907081 TI - Genetically modified HLA class I molecules able to inhibit human NK cells without provoking alloreactive CD8+ CTLs. AB - Human NK cells are likely to be important effectors of xenograft rejection. Expression of HLA class I molecules by transfected porcine cells can protect them from human NK cell-mediated lysis; however, this strategy has the potential to augment the anti-graft response by recipient CD8(+) T cells recognizing foreign pig peptides presented by HLA. In this study we show that the introduction of a mutation (D227K) in the alpha(3) domain of HLA-Cw3 abrogates its recognition by CD8-dependent T cells but leaves intact its ability to function as an inhibitory ligand for NK cells. Such genetically modified molecules may have potential therapeutic applications in the prevention of delayed xenograft rejection and in the facilitation of allogeneic and xenogeneic bone marrow engraftment. PMID- 11907082 TI - Intracellular domains of target antigens influence their capacity to trigger antibody-dependent cell-mediated cytotoxicity. AB - Ab-mediated signaling in tumor cells and Ab-dependent cell-mediated cytotoxicity (ADCC) are both considered as relevant effector mechanisms for Abs in tumor therapy. To address potential interactions between these two mechanisms, we generated HER-2/neu- and CD19-derived chimeric target Ags, which were expressed in experimental tumor target cells. HER-2/neu-directed Abs were documented to mediate effective ADCC with both mononuclear cells (MNCs) and polymorphonuclear granulocytes (PMNs), whereas Abs against CD19 were effective only with MNCs and not with PMNs. We generated cDNA encoding HER-2/CD19 or CD19/HER-2 (extracellular/intracellular) chimeric fusion proteins by combining cDNA encoding extracellular domains of HER-2/neu or CD19 with intracellular domains of CD19 or HER-2/neu, respectively. After transfecting wild-type HER-2/neu or chimeric HER 2/CD19 into Raji Burkitt's lymphoma cells and wild-type CD19 or chimeric CD19/HER 2 into SK-BR-3 breast cancer cells, target cell lines were selected for high membrane expression of transfected Ags. We then investigated the efficacy of tumor cell lysis by PMNs or MNCs with CD19- or HER-2/neu-directed Ab constructs. MNCs triggered effective ADCC against target cells expressing wild-type or chimeric target Ag. As expected, PMNs killed wild-type HER-2/neu-transfected, but not wild-type CD19-transfected target cells. Interestingly, however, PMNs were also effective against chimeric CD19/HER-2-transfected, but not HER-2/CD19 transfected target cells. In conclusion, these results demonstrate that intracellular domains of target Ags contribute substantially to effective Ab mediated tumor cell killing by PMNs. PMID- 11907083 TI - The cross-priming APC requires a Rel-dependent signal to induce CTL. AB - Induction of OVA-specific CTL by cross-priming requires help from CD4 T cells, which use CD154 to signal CD40 on the APC. To further dissect the molecular pathways involved in cross-priming, we examined the role of Rel, an NF-kappaB family member. c-rel(-/-) mice failed to generate OVA-specific CTL by cross priming, but could induce CTL to HSV-1. Using chimeric mice, Rel expression was shown to be required by the APC, but not by the T cells. Notably, the deficiency in Rel could be overcome by triggering CD40, implying that the APC required Rel before receipt of the CD40 signal. These data suggest that the cross-priming APC must receive two signals before it can stimulate CTL. The first signal is Rel dependent and is required before activation of CD4 helper T cells, which then deliver the second signal using CD154 to trigger CD40. PMID- 11907084 TI - Uncoupling of proliferation and Stat5 activation in thymic stromal lymphopoietin mediated signal transduction. AB - Thymic stromal lymphopoietin (TSLP) is a cytokine that facilitates B lymphocyte differentiation and costimulates T cells. Previous studies have demonstrated that a functional TSLP receptor complex is a heterodimer consisting of the TSLP receptor and the IL-7R alpha-chain. TSLP-mediated signaling is unique among members of the cytokine receptor family in that activation of the transcription factor Stat5 occurs without detectable Janus kinase activation. Using a variety of biological systems we demonstrate here that TSLP-mediated Stat5 activation can be uncoupled from proliferation. We also show that the single tyrosine residue in the cytoplasmic domain of the TSLP receptor is critical for TSLP-mediated proliferation, but is dispensable for Stat5 activation. Our data demonstrate that TSLP-mediated Stat5 activation is insufficient for cell proliferation and identifies residues within the TSLP receptor complex required to mediate these downstream events. PMID- 11907085 TI - Expression of inhibitory receptors Ly49E and CD94/NKG2 on fetal thymic and adult epidermal TCR V gamma 3 lymphocytes. AB - Ly49 and CD94/NKG2 inhibitory receptors are predominantly expressed on murine NK cells, but they are also expressed on a subpopulation of peripheral CD8 memory TCR alphabeta lymphocytes. In this study we demonstrate that Ly49E and CD94/NKG2 receptors are expressed on mature TCR Vgamma3(+) cells in the fetal thymus. Expression correlated with a memory phenotype, such as expression of CD44, 2B4, and IL-2Rbeta (CD122), and absence of IL-2Ralpha (CD25) expression. No expression of Ly49A, C, D, G2, or I receptors was observed. This phenotype is similar to that of fetal thymic NK cells. Skin-located Vgamma3 T cells, the progeny of fetal thymic Vgamma3 cells, also expressed CD94/NKG2 and Ly49E but not the other members of the Ly49 family. The development and survival of Ly49E(+) or CD94/NKG2(+) Vgamma3 T lymphocytes was not dependent upon expression of MHC class I molecules. The cytotoxicity of TCR Vgamma3 cells was inhibited when Qdm, the ligand for CD94/NKG2, was presented by Qa1(b)-transfected target cells. Also, upon cross-linking of CD94/NKG2 with mAb 3S9, TCR Vgamma3 thymocytes were prevented from killing FcgammaR(+) P815 target cells. These effects were most pronounced in the CD94/NKG2(high) subpopulation as compared with the CD94/NKG2(low) subpopulation of Vgamma3 cells. Our data demonstrate that Vgamma3 T cells expressing inhibitory Ly49E and CD94/NKG2 receptors are mature and display a memory phenotype, and that CD94/NKG2 functions as an inhibitory receptor on these T lymphocytes. PMID- 11907086 TI - Contribution of Langerhans cell-derived IL-18 to contact hypersensitivity. AB - The epidermal Langerhans cells (LC), a member of the dendritic cell family, and the LC-derived cytokine IL-12 play a pivotal role in the initiation of contact hypersensitivity (CHS), a Th1 immune response in the skin. Because IL-18, another LC-derived cytokine, shares functional and biological properties with IL-12, we examined a potential role for IL-18 in CHS initiation. Our studies demonstrated that during the induction phase of murine CHS, IL-18 mRNA was significantly up regulated in the skin-draining lymph nodes (LN). Migratory hapten-modified LC in LN expressed high levels of IL-18 mRNA and secreted functional IL-18 protein. LN cells produced significant amounts of IFN-gamma following in vitro IL-12 stimulation, which could be partially blocked by anti-IL-18 Ab, suggesting a synergistic role for endogenous IL-18 in IFN-gamma production by LN cells. Because mature IL-18 requires cleavage of immature precursors by caspase-1, we further examined IL-12-induced IFN-gamma production in caspase-1(-/-) LN cells. An impaired IFN-gamma production was seen in caspase-1(-/-) LN cells, which could be restored by addition of exogenous IL-18, supporting a role for caspase-1 cleaved, mature IL-18 in IFN-gamma production. Finally, in vivo studies showed that CHS responses were significantly inhibited in mice treated with neutralizing IL-18 Ab as well as in caspase-1(-/-) mice deficient in mature IL-18, indicating functional relevance for IL-18 in CHS. Taken together, our studies demonstrate that LC-derived IL-18 significantly contributes to CHS initiation. PMID- 11907087 TI - Clonal selection, clonal senescence, and clonal succession: the evolution of the T cell response to infection with a persistent virus. AB - We have analyzed the CD8(+) T cell response to EBV and find that a larger primary burst size is associated with proportionally greater decay during the development of memory. Consequently, immunodominance and clonal dominance are less marked in memory than primary responses. An intuitive interpretation of this finding is that there is a limit to the number of cell divisions a T cell clone can undergo, and that the progeny of clones that have expanded massively during a primary immune response are more prone to die as a result of senescence. To test this hypothesis, we have derived a mathematical model of the response of different T cell clones of varying avidity for Ag in the primary and persistent phases of viral infection. When cellular survival and replication are linked to T cell avidity for Ag and Ag dose, then high-avidity T cells dominate both the primary and secondary responses. We then incorporated a limit in the number of cell divisions of individual T cell clones to test whether such a constraint could reproduce the observed association between cell division number and alterations in the contribution of clones to the response to persistent infection. Comparison of the model output with the experimental results obtained from primary and persistent EBV infection suggests that there is indeed a role for cellular senescence in shaping the immune response to persistent infection. PMID- 11907088 TI - Role of TNF receptor-associated factor 2 in the activation of IgM secretion by CD40 and CD120b. AB - TNFR-associated factors (TRAFs) participate in the signaling of many TNFR family members, including CD40, CD120a (TNFR1), and CD120b (TNFR2). Previously, we found that a dominant-negative TRAF2 molecule inhibits CD40-mediated Ab secretion by the mouse B cell line CH12.LX. However, disruption of the TRAF2 binding site in the cytoplasmic domain of CD40 does not diminish the ability of CD40 to stimulate Ab secretion, nor is this mutation able to circumvent the inhibition of Ab secretion by dominant-negative TRAF2. Here we demonstrate that CD40-induced TNF stimulates IgM production through CD120b and that CD120b signaling is required for optimal CD40-induced IgM secretion. Furthermore, although both CD40 and CD120b can bind TRAF2, TRAF2-dependent CD40 signals cannot substitute for TRAF2 dependent CD120b signals in the activation of IgM secretion. Our results indicate a potentially important role for CD120b in the activation of IgM secretion and that TRAF2 is used by CD40 and CD120b in distinct ways. PMID- 11907089 TI - DQ 65-79, a peptide derived from HLA class II, induces I kappa B expression. AB - A synthetic peptide corresponding to residues 65-79 of the alpha helix of the alpha-chain of the class II HLA molecule DQA03011 (DQ 65-79) inhibits the proliferation of human T lymphocytes in an allele nonrestricted manner. By using microarray technology, we found that expression of 29 genes was increased or decreased in a human CTL cell line after treatment with DQ 65-79. This study focuses on one of these genes, IkappaB-alpha, whose expression is increased by DQ 65-79. IkappaB proteins, including IkappaB-alpha and IkappaB-beta, are increased in T cells treated with DQ 65-79. Nuclear translocation of the NF-kappaB subunits p65 and p50 is decreased in T cells after treatment with DQ 65-79, while elevated levels of p65 and p50 are present in cytosol. DQ 65-79 inhibits the degradation of IkappaB-alpha mRNA and inhibits the activity of IkappaB kinase. These findings indicate that the DQ 65-79 peptide increases the level of IkappaB proteins, thereby preventing nuclear translocation of the transcription factor, NF-kappaB, and inhibiting T cell proliferation. PMID- 11907090 TI - The first cytokine sequence within cartilaginous fish: IL-1 beta in the small spotted catshark (Scyliorhinus canicula). AB - Cartilaginous fish are considered the most primitive living jawed vertebrates with a complex immune system typical of all jawed vertebrates. Cytokine homologs are found within jawless and bony fish, although no cytokine or cytokine receptor genes have been sequenced in cartilaginous fish. In this study the complete coding sequence of the small spotted catshark (Scyliorhinus canicula) IL-1beta gene is presented that contains a short 5' untranslated region (54 bp), a 903-bp open reading frame, a 379-bp 3' untranslated region, a polyadenylation signal, and eight mRNA instability motifs. The predicted translation (301 amino acids) has highest identity to trout IL-1beta (31.7%), with greatest homology within the putative 12 beta-sheets. The IL-1 family signature is also present, but there is no apparent signal peptide. As with other nonmammalian IL-1beta sequences, the IL 1-converting enzyme cut site is absent. Expression of the IL-1beta transcript is detectable by RT-PCR in the spleen and testes, induced in vivo with LPS. Furthermore, a 7-fold increase of transcript levels in splenocytes incubated for 5 h with LPS was seen. The genomic organization comprises six exons and five introns with highest homology seen in exons encoding the largest amount of secondary structure per amino acid. Southern blot analysis suggests at least two copies of the IL-1beta gene or genes related to the 3' end of the IL-1beta sequence are present in the catshark. The cloning of IL-1beta in S. canicula, the first cytokine sequenced within cartilaginous fish, verifies previous bioactivity evidence for the presence of inflammatory cytokines. PMID- 11907091 TI - Yin Yang 1, Oct1, and NFAT-4 form repeating, cyclosporin-sensitive regulatory modules within the murine CD21 intronic control region. AB - The murine complement receptor type 2 gene (Cr2/CD21) is expressed by murine B and follicular dendritic cells, but not murine T cells. We have previously shown that appropriate transcriptional control of the CD21 gene requires the CD21 promoter as well as intronic sequences. We have also demonstrated that altering chromatin structure by inhibiting histone deacetylases induces CD21 expression in murine T cells by increasing the accessibility of promoter and intronic regulatory elements. In this report, we identify seven distinct regulatory areas within the first intron of the murine CD21 gene that are conserved between mouse and human CD21 intronic sequences. EMSA competition and supershift analyses reveal the formation of multiple DNA-protein complexes at these sites that include Yin Yang 1, Oct1, and NFAT-4. NFAT-containing complexes were altered in B cells treated with the NFAT inhibitor cyclosporin A and correlated with a repression of CD21 gene transcription implicating NFAT transcriptional control. Functional data revealed that no single region conferred cell-specific reporter gene expression, but rather the entire CD21 regulatory element was required to confer cell-specific gene expression. Taken together, these data demonstrate the formation of repeating, overlapping regulatory modules, all of which are required to coordinately control the cell-specific expression of the murine CD21 gene. We propose a model in which Yin Yang 1 and Oct1 may recruit histone deacetylase to multiple sites in the CD21 intronic regulatory element in nonexpressing cells and NFAT either displaces this histone deacetylase or recruits a histone acetylase to allow the formation of a functional transcriptional complex in expressing cells. PMID- 11907092 TI - Mutational analysis of immunoreceptor tyrosine-based inhibition motifs of the Ig like transcript 2 (CD85j) leukocyte receptor. AB - The inhibitory receptor Ig-like transcript (ILT)2 (leukocyte Ig-like receptor or CD85j) is a type I transmembrane protein expressed by different leukocyte lineages. The extracellular region of ILT2 binds HLA class I molecules, and its cytoplasmic domain displays four immunoreceptor tyrosine-based inhibition motifs. Upon tyrosine phosphorylation ILT2 recruits the Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP-1) that is involved in negative signaling. To address the structural basis of ILT2-mediated inhibitory signaling, deletion and single tyrosine mutants were generated and transfected in the COS-7 and rat basophilic leukemia cell lines; their abilities to bind SHP-1 and to inhibit FcepsilonR-induced serotonin release in rat basophilic leukemia cells were studied. Both biochemical and functional analyses revealed tyrosines 644 (SIYATL) and 614 (VTYAQL) as the SHP-1 docking sites required for ILT2 inhibitory function. Substitution of tyrosine 562 (VTYAEV) did not alter receptor function. By contrast, mutation of tyrosine 533 (NLYAAV) interfered with ILT2 tyrosine phosphorylation and the subsequent SHP-1 recruitment, thus supporting a regulatory role for this motif. PMID- 11907093 TI - Functional analysis of I alpha promoter regions of multiple IgA heavy chain genes. AB - The 13 nonallelic IgA H chain genes of rabbit are differentially expressed in vivo. They can be grouped into those expressed at high levels (Calpha4, Calpha5, Calpha6, Calpha9, Calpha10, Calpha12, and Calpha13), those expressed at low levels (Calpha1, Calpha2, Calpha7, and Calpha11), and those that are not expressed (Calpha3 and Calpha8). We tested whether the differential in vivo expression is due to differential responses of the Ialpha promoters to TGF-beta stimulation. We stimulated the rabbit B cell line 55D1 with TGF-beta and, using single-cell RT-PCR, found that expression of germline (GL) transcripts of alpha3 and alpha8 could not be induced. By luciferase reporter gene assay and EMSA we found that the promoters of the unexpressed isotypes Calpha3 and Calpha8 are defective, thereby explaining the absence of IgA3 and IgA8 in vivo. When comparing the promoter activities of the other isotypes we found that the activities did not reflect the degree of in vivo expression. Instead, the promoters of the isotypes expressed at high or low levels promoted expression of the luciferase gene to a similar degree, except for the Ialpha4 promoter, which had much higher activity. Also the degree to which TGF-beta induced GL expression of the various isotypes in 55D1 B cells did not reflect in vivo expression. However, most of the TGF-beta-stimulated cells expressed GL mRNA of multiple isotypes; no isotype was expressed preferentially. These results suggest that the final switch to a single isotype is regulated in a step subsequent to GL transcription, rather than by induction of GL transcripts by the Ialpha promoter. PMID- 11907094 TI - Bob1 (OCA-B/OBF-1) differential transactivation of the B cell-specific B29 (Ig beta) and mb-1 (Ig alpha) promoters. AB - The B29 (Igbeta) and mb-1 (Igalpha) gene products are B cell-specific essential components of the B cell receptor that are coexpressed at all stages of B cell differentiation, with the exception of plasma cells, which lack mb-1 expression. Transcription of both genes is governed by a similar cassette of interactive transcription factor-binding elements, including octamer motifs, in TATA-less promoters. In this study, we show the B cell-specific B29 gene promoter is transactivated in B and non-B cells by cotransfection with the B cell-specific octamer cofactor gene, Bob1 (OCA-B/OBF-1). The expression of Bob1 is also sufficient to override the silencing effects of the B29 silencer. This indicates that Bob1 plays a critical role in B cell-specific B29 promoter expression. In contrast, coexpression of Bob1 had no effect on mb-1 promoter activity. Bob1 transactivation only occurs with select octamer sequences that have an adenosine at position 5 (ATGCAAAT). The B29 promoter conforms to this consensus octamer motif, while the mb-1 promoter octamer motif does not. Octamer motif swapping between B29 and mb-1 promoters renders B29 unresponsive to Bob1 transactivation and makes mb-1 competent for Bob1 transactivation, thereby indicating that the B29 octamer motif is solely responsible for Bob1 interaction. Additionally, the mb-1 construct containing the B29 octamer motif is expressed in a plasmacytoma cell line, while the wild-type mb-1 promoter is not. Bob1 transactivation of B29 and the lack of this transactivation of mb-1 account for the differential expression of B29 and mb-1 in terminally differentiated plasma cells. PMID- 11907095 TI - The Rac2 guanosine triphosphatase regulates B lymphocyte antigen receptor responses and chemotaxis and is required for establishment of B-1a and marginal zone B lymphocytes. AB - We have defined roles for the hemopoietic-specific Rho guanosine triphosphatase, Rac2, in B lymphocyte development and function through examination of rac2(-/-) mice. Rac2-deficient mice displayed peripheral blood B lymphocytosis and marked reductions in peritoneal cavity B-1a lymphocytes, marginal zone B lymphocytes, and IgM-secreting plasma cells as well as reduced concentrations of serum IgM and IgA. The rac2(-/-) B lymphocytes exhibited reduced calcium flux following coligation of B cell AgR and CD19 and reduced chemotaxis in chemokine gradients. T cell-independent responses to DNP-dextran were of reduced magnitude, but normal kinetics, in rac2(-/-) mice, while T-dependent responses to nitrophenyl-keyhole limpet hemocyanin were subtly abnormal. Rac2 is therefore an essential element in regulating B lymphocyte functions and maintaining B lymphocyte populations in vivo. PMID- 11907096 TI - Segregation of Bad from lipid rafts is implicated in the induction of apoptosis. AB - Many molecules relocate subcellularly in cells undergoing apoptosis. Using coimmunoprecipitation experiments we demonstrate that Bad is not associated to 14 3-3 protein, suggesting a new mechanism for the control of the proapoptotic role of Bad. Here we show, by confocal microscopy and cellular fractionation, that Bad is attached to lipid rafts in IL-4-stimulated cells and thymocytes while associated with mitochondria in IL-4-deprived cells. Disruption of lipid rafts by methyl-beta-cyclodextrin treatment induces segregation of Bad from rafts, which correlates with apoptosis. Our results suggest that the interaction of Bad with rafts is a dynamic process regulated by IL-4 and involved in the control of apoptosis. PMID- 11907097 TI - Transmembrane TNF induces an efficient cell-mediated immunity and resistance to Mycobacterium bovis bacillus Calmette-Guerin infection in the absence of secreted TNF and lymphotoxin-alpha. AB - The contribution of a transmembrane (Tm) form of TNF to protective immunity against Mycobacterium bovis bacillus Calmette-Guerin (BCG) was studied in transgenic (tg) mice expressing a noncleavable Tm TNF but lacking the TNF/lymphotoxin-alpha (LT-alpha) locus (Tm TNF tg mice). These mice were as resistant to BCG infection as wild-type mice, whereas TNF/LT-alpha(-/-), TNF(-/ ), and LT-alpha(-/-) mice succumbed. Tm TNF tg mice developed granulomas of smaller size but at 2- to 4-fold increased frequencies compared with wild-type mice. Granulomas were mainly formed by monocytes and activated macrophages expressing Tm TNF mRNA and accumulating acid phosphatase. NO synthase 2 activation as a key macrophage bactericidal mechanism was low during the acute phase of infection in Tm TNF tg mice but was still sufficient to limit bacterial growth and increased in late infection. While infection with virulent Mycobacterium tuberculosis resulted in very rapid death of TNF/LT-alpha(-/-) mice, it also resulted in survival of Tm TNF tg mice which presented an increase in the number of CFU in spleen (5-fold) and lungs (10-fold) as compared with bacterial load of wild-type mice. In conclusion, the Tm form of TNF induces an efficient cell-mediated immunity and total resistance against BCG even in the absence of LT-alpha and secreted TNF. However, Tm TNF-mediated protection against virulent M. tuberculosis infection can also be efficient but not as strong as in BCG infection, in which cognate cellular interactions may play a more predominant role in providing long-term surveillance and containment of BCG-infected macrophages. PMID- 11907098 TI - Autocrine deactivation of macrophages in transgenic mice constitutively overexpressing IL-10 under control of the human CD68 promoter. AB - IL-10 plays an essential role in blocking cytokine production by activated macrophages. To analyze the consequences of enforced expression of IL-10 by macrophages on innate and adaptive immune responses, we generated transgenic mice (macIL-10tg mice) expressing an epitope-tagged IL-10 (Flag-IL-10) under control of the human CD68 promoter. Expression of Flag-IL-10 was constitutive and restricted to macrophages, as shown by sorting splenocyte cell populations and intracellular staining for IL-10. Transgenic macrophages displayed suppressed production of TNF-alpha and IL-12 upon stimulation with LPS. When macIL-10tg mice were challenged with LPS, serum levels of proinflammatory cytokines were attenuated compared with controls. Infection with Mycobacterium bovis bacille Calmette-Guerin resulted in approximately 10-fold-higher bacterial loads than in wild-type mice. Normal T and B cell responses were observed in macIL-10tg mice, suggesting that macrophage-specific overexpression of IL-10 predominantly acts in an autocrine/paracrine manner, resulting in chronically deactivated macrophages that manifest an impaired ability to control pathogens. PMID- 11907099 TI - Increased survival in sepsis by in vivo adenovirus-induced expression of IL-10 in dendritic cells. AB - The dendritic cell (DC) is the most potent APC of the immune system, capable of stimulating naive T cells to proliferate and differentiate into effector T cells. Recombinant adenovirus (Adv) readily transduces DCs in vitro allowing directed delivery of transgenes that modify DC function and immune responses. In this study we demonstrate that footpad injection of a recombinant Adv readily targets transduction of myeloid and lymphoid DCs in the draining popliteal lymph node, but not in other lymphoid organs. Popliteal DCs transduced with an empty recombinant Adv undergo maturation, as determined by high MHC class II and CD86 expression. However, transduction with vectors expressing human IL-10 limit DC maturation and associated T cell activation in the draining lymph node. The extent of IL-10 expression is dose dependent; transduction with low particle numbers (10(5)) yields only local expression, while transduction with higher particle numbers (10(7) and 10(10)) leads additionally to IL-10 appearance in the circulation. Furthermore, local DC expression of human IL-10 following in vivo transduction with low particle numbers (10(5)) significantly improves survival following cecal ligation and puncture, suggesting that compartmental modulation of DC function profoundly alters the sepsis-induced immune response. PMID- 11907100 TI - Antibody efficacy in murine pulmonary Cryptococcus neoformans infection: a role for nitric oxide. AB - We investigated the pathogenesis of pulmonary Cryptococcus neoformans infection and passive Ab efficacy in mice deficient in inducible NO synthase (NOS2(-/-)) and the parental strain. Parental mice lived significantly longer than NOS2(-/-) mice after intratracheal infection, despite having a higher lung fungal burden. Administration of Ab reduced lung CFU in both NOS2(-/-) and parental mice, but prolonged survival and increased the inflammatory response only in parental mice. Ab administration was associated with increased serum nitrite and reduced polysaccharide levels in parental mice. Eosinophils were present in greater numbers in the lung of infected NOS2(-/-) mice than parental mice, irrespective of Ab administration. C. neoformans infection in NOS2(-/-) mice resulted in significantly higher levels of IFN-gamma, monocyte chemoattractant protein-1, and macrophage-inflammatory protein-1alpha than parental mice. Ab administration had different effects on infected NOS2(-/-) and parental mice with respect to IFN gamma, monocoyte chemoattractant protein-1, and macrophage-inflammatory protein 1alpha levels. Ab administration increased lung levels of IFN-gamma in parental mice and reduced levels in NOS2(-/-) mice. The results indicate that NO is involved in the regulation of cytokine expression in response to cryptococcal pneumonia and is necessary for Ab efficacy against C. neoformans in mice. Our findings indicate a complex relationship between Ab efficacy against C. neoformans and cytokine expression, underscoring the interdependency of cellular and humoral defense mechanisms. PMID- 11907101 TI - The IFN-inducible Golgi- and endoplasmic reticulum- associated 47-kDa GTPase IIGP is transiently expressed during listeriosis. AB - Members of the 47-kDa GTPase family are implicated in an IFN-gamma-induced, as yet unclear, mechanism that confers innate resistance against infection with intracellular pathogens. Overt immunological parameters are apparently uncompromised in mice deficient for individual members and the prototype of this family, IGTP, localizes to the endoplasmic reticulum. This suggests that these GTPases are involved in intracellular defense. We analyzed the expression of the 47-kDa GTPase cognate, IIGP, in splenic sections from mice infected with the intracellular pathogen Listeria monocytogenes by immunohistochemistry. An early transient IIGP induction was observed revealing the IFN-gamma responsiveness of cellular subcompartments within the spleen in early listeriosis. Marginal metallophilic macrophages and endothelial cells within the red and white pulp strongly expressed IIGP, while other splenocytes remained negative. In vitro analyses show that both type I and type II IFNs are prime stimuli for IIGP induction in various cells, including L. monocytogenes-infected or LPS-stimulated macrophages, endothelial cells, and activated T cells. Contrary to the subcellular localization of IGTP, IIGP was predominantly associated with the Golgi apparatus and also localizes to the endoplasmic reticulum. We conclude that IIGP exerts a distinct role in IFN-induced intracellular membrane trafficking or processing. PMID- 11907102 TI - Gamma 3 gene-disrupted mice selectively deficient in the dominant IgG subclass made to bacterial polysaccharides. II. Increased susceptibility to fatal pneumococcal sepsis due to absence of anti-polysaccharide IgG3 is corrected by induction of anti-polysaccharide IgG1. AB - Bacterial polysaccharides (PS) are type 2 T-independent Ags that elicit Abs restricted in isotype to IgM and predominantly IgG2 in humans and IgM, and IgG3 in mice. Humans with IgG2 subclass deficiency are susceptible to sinus and pulmonary infections with PS-encapsulated bacteria. We previously developed an IgG3-deficient mouse by disrupting the gamma3 H chain constant region gene via targeted mutagenesis. Mutant mice lacking IgG3 were backcrossed for 10 generations to wild-type (WT) BALB/c mice to generate BALB/c mice that have complete absence of IgG3. WT mice immunized with type 3 Streptococcus pneumoniae capsular PS made anti-PS IgM, IgG3, and small quantities of IgG1, which opsonized S. pneumoniae for killing by polymorphonuclear leukocytes. These mice were protected against death from lethal doses of type 3 S. pneumoniae. In contrast, IgG3(-/-) mice made similar titers of anti-PS IgM and IgG1 as WT mice but no IgG3, and had poorly opsonic sera with significantly increased mortality after S. pneumoniae challenge. Immunization of IgG3(-/-) mice with type 3 S. pneumoniae PS conjugated to carrier protein CRM(197)-elicited IgM and high-titer IgG1 Abs, restored serum opsonization, and gave protection from mortality after S. pneumoniae, challenge comparable to WT mice. We conclude that mice lacking the dominant IgG3 subclass made to bacterial PS are more susceptible to fatal S. pneumoniae sepsis than WT mice, but that IgG1 induced by a S. pneumoniae glycoconjugate can adequately protect against S. pneumoniae sepsis. This model suggests that IgG subclass of anti-PS Ab is an important component of immunity to encapsulated bacteria. PMID- 11907103 TI - Plasmodium falciparum variant surface antigen expression varies between isolates causing severe and nonsevere malaria and is modified by acquired immunity. AB - In areas of endemic parasite transmission, protective immunity to Plasmodium falciparum malaria is acquired over several years with numerous disease episodes. Acquisition of Abs to parasite-encoded variant surface Ags (VSA) on the infected erythrocyte membrane is important in the development of immunity, as disease causing parasites appear to be those not controlled by preexisting VSA-specific Abs. In this work we report that VSA expressed by parasites from young Ghanaian children with P. falciparum malaria were commonly and strongly recognized by plasma Abs from healthy children in the same area, whereas recognition of VSA expressed by parasites from older children was weaker and less frequent. Independent of this, parasites isolated from children with severe malaria (cerebral malaria and severe anemia) were better recognized by VSA-specific plasma Abs than parasites obtained from children with nonsevere disease. This was not due to a higher infection multiplicity in younger patients or in patients with severe disease. Our data suggest that acquisition of VSA-specific Ab responses gradually restricts the VSA repertoire that is compatible with parasite survival in the semi-immune host. This appears to limit the risk of severe disease by discriminating against the expression of VSA likely to cause life threatening complications, such as cerebral malaria and severe anemia. Such VSA seem to be preferred by parasites infecting a nonimmune host, suggesting that VSA expression and switching are not random, and that the VSA expression pattern is modulated by immunity. This opens the possibility of developing morbidity reducing vaccines targeting a limited subset of common and particularly virulent VSA. PMID- 11907104 TI - The NKp46 receptor contributes to NK cell lysis of mononuclear phagocytes infected with an intracellular bacterium. AB - We used human tuberculosis as a model to investigate the role of NK cytotoxic mechanisms in the immune response to intracellular infection. Freshly isolated NK cells and NK cell lines from healthy donors lysed Mycobacterium tuberculosis infected monocytes to a greater extent than uninfected monocytes. Lysis of infected monocytes was associated with increased expression of mRNA for the NKp46 receptor, but not the NKp44 receptor. Antisera to NKp46 markedly inhibited lysis of infected monocytes. NK cell-mediated lysis was not due to reduced expression of MHC class I molecules on the surface of infected monocytes or to enhanced production of IL-18 or IFN-gamma. NK cell lytic activity against M. tuberculosis infected monocytes and NKp46 mRNA expression were reduced in tuberculosis patients with ineffective immunity to M. tuberculosis compared with findings in healthy donors. These observations suggest that 1) the NKp46 receptor participates in NK cell-mediated lysis of cells infected with an intracellular pathogen, and 2) the reduced functional capacity of NK cells is associated with severe manifestations of infectious disease. PMID- 11907105 TI - Iron chelation via deferoxamine exacerbates experimental salmonellosis via inhibition of the nicotinamide adenine dinucleotide phosphate oxidase-dependent respiratory burst. AB - Competition for cellular iron (Fe) is a vital component of the interaction between host and intracellular pathogen. The host cell requires Fe for the execution of antimicrobial effector mechanisms, whereas most bacteria have an obligate requirement for Fe to sustain growth and intracellular survival. In this study, we show that chelation of host Fe in vivo exacerbates murine salmonellosis, resulting in increased bacterial load and decreased survival times. We further demonstrate that host Fe deprivation results in an inability to induce the NADPH oxidase-dependent production of reactive oxygen, an essential host defense mechanism for the early control of Salmonella typhimurium infection. Thus, altering the equilibrium of intracellular Fe influences the course of infection to the benefit of the pathogen. PMID- 11907106 TI - The MHC class I homolog of human cytomegalovirus is resistant to down-regulation mediated by the unique short region protein (US)2, US3, US6, and US11 gene products. AB - Human CMV encodes four unique short region proteins (US), US2, US3, US6, and US11, each independently sufficient for causing the down-regulation of MHC class I molecules on the cell surface. This down-regulation allows infected cells to evade recognition by cytotoxic T cells but leaves them susceptible to NK cells, which lyse cells that lack class I molecules. Another human CMV-encoded protein, unique long region protein 18 (UL18), is an MHC class I homolog that might provide a mechanism for inhibiting the NK cell response. The sequence similarities between MHC class I molecules and UL18 along with the ability of UL18 to form trimeric complexes with beta(2)-microglobulin and peptides led to the hypothesis that if the US and UL18 gene products coexist temporally during infection, the US proteins might down-regulate UL18 molecules, similar to their action on MHC class I molecules. We show here that temporal expression of US and UL18 genes partially overlaps during infection. However, unlike MHC class I molecules, the MHC class I homolog, UL18, is fully resistant to the down regulation associated with the US2, US3, US6, and US11 gene products. The specific effect of US proteins on MHC class I molecules, but not on UL18, represents another example of how viral proteins have evolved to evade immune surveillance, avoiding fratricide by specifically targeting host proteins. PMID- 11907107 TI - Preferential infection of immature dendritic cells and B cells by mouse mammary tumor virus. AB - Until now it was thought that the retrovirus mouse mammary tumor virus preferentially infects B cells, which thereafter proliferate and differentiate due to superantigen-mediated T cell help. We describe in this study that dendritic cells are infectable at levels comparable to B cells in the first days after virus injection. Moreover, IgM knockout mice have chronically deleted superantigen-reactive T cells after MMTV injection, indicating that superantigen presentation by dendritic cells is sufficient for T cell deletion. In both subsets initially only few cells were infected, but there was an exponential increase in numbers of infected B cells due to superantigen-mediated T cell help, explaining that at the peak of the response infection is almost exclusively found in B cells. The level of infection in vivo was below 1 in 1000 dendritic cells or B cells. Infection levels in freshly isolated dendritic cells from spleen, Langerhans cells from skin, or bone marrow-derived dendritic cells were compared in an in vitro infection assay. Immature dendritic cells such as Langerhans cells or bone marrow-derived dendritic cells were infected 10- to 30-fold more efficiently than mature splenic dendritic cells. Bone marrow-derived dendritic cells carrying an endogenous mouse mammary tumor virus superantigen were highly efficient at inducing a superantigen response in vivo. These results highlight the importance of professional APC and efficient T cell priming for the establishment of a persistent infection by mouse mammary tumor virus. PMID- 11907108 TI - Reduced functional capacity of CD8+ T cells expanded by post-exposure vaccination of gamma-herpesvirus-infected CD4-deficient mice. AB - Mice (I-A(b-/-)) that lack CD4(+) T cells remain healthy for at least three months after respiratory exposure to the murine gamma-herpesvirus 68 (gammaHV68), then succumb with symptoms of chronic wasting disease. Postexposure challenge of gammaHV68-infected I-A(b+/+) and I-A(b-/-) mice with a recombinant vaccinia virus (Vacc-p56) expressing an antigenic gammaHV68 peptide caused a massive increase in the numbers of D(b)p56-specific CD8(+) T cells. Previous experiments showed that, despite the large numbers of potential CTL effectors, there was little effect on the long-term survival of the CD4-deficient group and no diminution in the level of persistent virus shedding and latency. Comparison of the expanded CD8(+)D(b)p56(+) sets in the I-A(b+/+) and I-A(b-/-) mice indicated that these two T cell populations were not identical. More CD69(high)CD8(+) D(b)p56(+) T cells were found in the CD4-deficient mice, an effect that might be thought to reflect higher Ag load. By contrast, the mean fluorescence intensity of staining for the CD44 glycoprotein was diminished on CD8(+)D(b)p56(+) T cells from the I A(b-/-) group, the level of CTL activity was lower on a per cell basis, and the relative prevalence of IFN-gamma(+)TNF-alpha(+) T cells detected after in vitro stimulation with the p56 peptide was decreased. Given that this experimental system provides an accessible model for evaluating postexposure vaccination protocols that might be used in diseases like HIV/AIDS, the further need is to clarify the underlying molecular mechanisms and the relative significance of lack of CD4(+) T help vs higher Ag load for these expanded CD8(+) effector populations. PMID- 11907109 TI - Tumor-specific CTL kill murine renal cancer cells using both perforin and Fas ligand-mediated lysis in vitro, but cause tumor regression in vivo in the absence of perforin. AB - Kidney cancer is a devastating disease; however, biological therapies have achieved some limited success. The murine renal cancer Renca has been used as a model for developing new preclinical approaches to the treatment of renal cell carcinoma. Successful cytokine-based approaches require CD8(+) T cells, but the exact mechanisms by which T cells mediate therapeutic benefit have not been completely identified. After successful biological therapy of Renca in BALB/c mice, we generated CTLs in vitro using mixed lymphocyte tumor cultures. These CTL mediated tumor-specific H-2K(d)-restricted lysis and production of IFN-gamma, TNF alpha, and Fas ligand (FasL) in response to Renca. CTL used both granule- and FasL-mediated mechanisms to lyse Renca, although granule-mediated killing was the predominant lytic mechanism in vitro. The cytokines IFN-gamma and TNF-alpha increased the sensitivity of Renca cells to CTL lysis by both granule- and FasL mediated death pathways. Adoptive transfer of these anti-Renca CTL into tumor bearing mice cured most mice of established experimental pulmonary metastases, and successfully treated mice were immune to tumor rechallenge. Interestingly, we were able to establish Renca-specific CTL from mice gene targeted for perforin (pfp(-/-)) mice. Although these pfp(-/-) CTL showed reduced cytotoxic activity against Renca, their IFN-gamma production in the presence of Renca targets was equivalent to that of wild-type CTL, and adoptive transfer of pfp(-/-) CTL was as efficient as wild-type CTL in causing regression of established Renca pulmonary metastases. Therefore, although granule-mediated killing is of paramount importance for CTL-mediated lysis in vitro, some major in vivo effector mechanisms clearly are independent of perforin. PMID- 11907110 TI - Depletion of CD8+ T cells exacerbates CD4+ Th cell-associated inflammatory lesions during murine mycoplasma respiratory disease. AB - Mycoplasma infection is a leading cause of pneumonia worldwide and can lead to other respiratory complications. A component of mycoplasma respiratory diseases is immunopathologic, suggesting that lymphocyte activation is a key event in the progression of these chronic inflammatory diseases. The present study delineates the changes in T cell populations and their activation after mycoplasma infection and determines their association with the pathogenesis of murine Mycoplasma respiratory disease, due to Mycoplasma pulmonis infection. Increases in T cell population numbers in lungs and lower respiratory lymph nodes were associated with the development of mycoplasma respiratory disease. Although both pulmonary Th and CD8(+) T cells increased after mycoplasma infection, there was a preferential expansion of Th cells. Mycoplasma-specific Th2 responses were dominant in lower respiratory lymph nodes, while Th1 responses predominated in spleen. However, both mycoplasma-specific Th1 and Th2 cytokine (IL-4 and IFN gamma) responses were present in the lungs, with Th1 cell activation as a major component of the pulmonary Th cell response. Although a smaller component of the T cell response, mycoplasma-specific CD8(+) T cells were also a significant component of pulmonary lymphoid responses. In vivo depletion of CD8(+) T cells resulted in dramatically more severe pulmonary disease, while depletion of CD4(+) T cells reduced its severity, but there was no change in mycoplasma numbers in lungs after cell depletion. Thus, mycoplasma-specific Th1 and CD8(+) T cell activation in the lung plays a critical regulatory role in development of immunopathologic reactions in Mycoplasma respiratory disease. PMID- 11907111 TI - Activation of the lectin complement pathway by H-ficolin (Hakata antigen). AB - Ficolins are a group of proteins which consist of a collagen-like domain and a fibrinogen-like domain. In human serum, there are two types of ficolins named L ficolin/P35 and H-ficolin (Hakata Ag), both of which have lectin activity. We recently demonstrated that L-ficolin/P35 is associated with mannose-binding lectin (MBL)-associated serine proteases (MASP) 1 and 2 and small MBL-associated protein (sMAP), and that the complex activates the lectin pathway. In this study, we report the characterization of H-ficolin in terms of its ability to activate complement. Western blotting analysis showed the presence of MASP-1, MASP-2, MASP 3, and sMAP in H-ficolin preparations isolated from Cohn Fraction III. The MASPs in the preparations had proteolytic activities against C4, C2, and C3 in the fluid phase. When H-ficolin preparations were bound to anti-H-ficolin Ab which had been coated on ELISA plates, they activated C4, although no C4 activation was noted when anti-MBL and anti-L-ficolin/P35 were used. H-ficolin binds to PSA, a polysaccharide produced by Aerococcus viridans. C4 was activated by H-ficolin preparations bound to PSA which had been coated on ELISA plates. These results indicate that H-ficolin is a second ficolin which is associated with MASPs and sMAP, and which activates the lectin pathway. PMID- 11907112 TI - Urokinase receptor is necessary for adequate host defense against pneumococcal pneumonia. AB - Cell recruitment is a multistep process regulated by cytokines, chemokines, and growth factors. Previous work has indicated that the urokinase plasminogen activator receptor (uPAR) may also play a role in this mechanism, presumably by an interaction with the beta(2) integrin CD11b/CD18. Indeed, an essential role of uPAR in neutrophil recruitment during pulmonary infection has been demonstrated for beta(2) integrin-dependent respiratory pathogens. We investigated the role of uPAR and urokinase plasminogen activator (uPA) during pneumonia caused by a beta(2) integrin-independent respiratory pathogen, Streptococcus pneumoniae. uPAR deficient (uPAR(-/-)), uPA-deficient (uPA(-/-)), and wild-type (Wt) mice were intranasally inoculated with 10(5) CFU S. pneumoniae. uPAR(-/-) mice showed reduced granulocyte accumulation in alveoli and lungs when compared with Wt mice, which was associated with more S. pneumoniae CFU in lungs, enhanced dissemination of the infection, and a reduced survival. In contrast, uPA(-/-) mice showed enhanced host defense, with more neutrophil influx and less pneumococci in the lungs compared with Wt mice. These data suggest that uPAR is necessary for adequate recruitment of neutrophils into the alveoli and lungs during pneumonia caused by S. pneumoniae, a pathogen eliciting a beta(2) integrin-independent inflammatory response. This function is even more pronounced when uPAR is unoccupied by uPA. PMID- 11907113 TI - Tumor regression induced by intratumor therapy with a disabled infectious single cycle (DISC) herpes simplex virus (HSV) vector, DISC/HSV/murine granulocyte macrophage colony-stimulating factor, correlates with antigen-specific adaptive immunity. AB - Direct intratumor injection of a disabled infectious single cycle HSV-2 virus encoding the murine GM-CSF gene (DISC/mGM-CSF) into established murine colon carcinoma CT26 tumors induced a significant delay in tumor growth and complete tumor regression in up to 70% of animals. Pre-existing immunity to HSV did not reduce the therapeutic efficacy of DISC/mGM-CSF, and, when administered in combination with syngeneic dendritic cells, further decreased tumor growth and increased the incidence of complete tumor regression. Direct intratumor injection of DISC/mGM-CSF also inhibited the growth of CT26 tumor cells implanted on the contralateral flank or seeded into the lungs following i.v. injection of tumor cells (experimental lung metastasis). Proliferation of splenocytes in response to Con A was impaired in progressor and tumor-bearer, but not regressor, mice. A potent tumor-specific CTL response was generated from splenocytes of all mice with regressing, but not progressing tumors following in vitro peptide stimulation; this response was specific for the gp70 AH-1 peptide SPSYVYHQF and correlated with IFN-gamma, but not IL-4 cytokine production. Depletion of CD8(+) T cells from regressor splenocytes before in vitro stimulation with the relevant peptide abolished their cytolytic activity, while depletion of CD4(+) T cells only partially inhibited CTL generation. Tumor regression induced by DISC/mGM-CSF virus immunotherapy provides a unique model for evaluating the immune mechanism(s) involved in tumor rejection, upon which tumor immunotherapy regimes may be based. PMID- 11907114 TI - Reduced expression of nuclear cyclic adenosine 5'-monophosphate response element binding proteins and IFN-gamma promoter function in disease due to an intracellular pathogen. AB - Mycobacterium tuberculosis-induced IFN-gamma protein and mRNA expression have been shown to be reduced in tuberculosis patients, compared with healthy tuberculin reactors. To determine whether this decrease was associated with reduced activity of the IFN-gamma promoter, we first studied binding of nuclear proteins to the radiolabeled proximal IFN-gamma promoter (-71 to -40 bp), using EMSAs with nuclear extracts of freshly isolated peripheral blood T cells. Nuclear extracts of T cells from most tuberculosis patients showed markedly reduced expression of proteins that bind to the proximal IFN-gamma promoter, compared with findings in nuclear extracts of T cells from healthy tuberculin reactors. These DNA-binding complexes contained CREB proteins, based on competitive EMSAs, supershift assays, and Western blotting with an anti-CREB Ab. Transient transfection of PBLs with a luciferase reporter construct under the control of the IFN-gamma promoter revealed reduced IFN-gamma promoter activity in tuberculosis patients. Transient transfection of Jurkat cells with a dominant negative CREB repressor plasmid reduced IFN-gamma promoter activity. These data suggest that reduced expression of CREB nuclear proteins in tuberculosis patients results in decreased IFN-gamma promoter activity and reduced IFN-gamma production. PMID- 11907116 TI - The proinflammatory CD14+CD16+DR++ monocytes are a major source of TNF. AB - In human blood two monocyte populations can be distinguished, i.e., the CD14(++)CD16(-)DR(+) classical monocytes and the CD14(+)CD16(+)DR(++) proinflammatory monocytes that account for only 10% of all monocytes. We have studied TNF production in these two types of cells using three-color immunofluorescence and flow cytometry on whole peripheral blood samples stimulated with either LPS or with the bacterial lipopeptide S-(2,3 bis(palmitoyloxy)-(2-RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser-(S)-Lys(4) OH,trihydrochloride (Pam3Cys). After stimulation with LPS the median fluorescence intensity for TNF protein was 3-fold higher in the proinflammatory monocytes when compared with the classical monocytes. After stimulation with Pam3Cys they almost exclusively responded showing 10-fold-higher levels of median fluorescence intensity for TNF protein. The median fluorescence intensity for Toll-like receptor 2 cell surface protein was found 2-fold higher on CD14(+)CD16(+)DR(++) monocytes, which may explain, in part, the higher Pam3Cys-induced TNF production by these cells. When analyzing secretion of TNF protein into the supernatant in PBMCs after depletion of CD16(+) monocytes we found a reduction of LPS-induced TNF by 28% but Pam3Cys-induced TNF was reduced by 64%. This indicates that the minor population of CD14(+)CD16(+) monocytes are major producers of TNF in human blood. PMID- 11907115 TI - Soluble fibrinogen modulates neutrophil functionality through the activation of an extracellular signal-regulated kinase-dependent pathway. AB - The integrin family not only mediates the recruitment of polymorphonuclear leukocytes (PMN) to sites of inflammation but also regulates several effector functions by binding to specific ligands. We have recently demonstrated that soluble fibrinogen (sFbg) is able to trigger an activating signal in PMN through an integrin-dependent mechanism. This activation results in degranulation, phagocytosis enhancement, and apoptosis delay. The aim of the present work was to further elucidate the molecular events that follow sFbg interaction with CD11b in human PMN, and the participation of this signaling pathway in the regulation of neutrophil functionality. We demonstrate that sFbg triggers a cascade of intracellular signals that lead to focal adhesion kinase and extracellular signal regulated kinase 1/2 tyrosine phosphorylation. The activation of this mitogen activated protein kinase pathway plays a central role in the sFbg modulation of secondary granule degranulation, Ab-dependent phagocytosis, and apoptosis. However, fibrinogen-induced secretory vesicle degranulation occurs independently of the signaling transduction pathways investigated herein. In the context of an inflammatory process, the intracellular signal pathway activated by sFbg may be an early event influencing the functionality of PMN. PMID- 11907117 TI - Gq signaling is required for allergen-induced pulmonary eosinophilia. AB - The complexity and magnitude of interactions leading to the selective infiltration of eosinophils in response to inhaled allergens are formidable obstacles to a larger understanding of the pulmonary pathology associated with allergic asthma. This study uses knockout mice to demonstrate a novel function for the heterotrimeric G protein, G(q), in the regulation of pulmonary eosinophil recruitment. In the absence of G(q) signaling, eosinophils failed to accumulate in the lungs following allergen challenge. These studies demonstrate that the inhibition of eosinophil accumulation in the airways is attributed to the failure of hemopoietically derived cells to elaborate GM-CSF in the airways. The data suggest that activation of a G(q)-coupled receptor(s) on resident leukocytes in the lung elicits expression of GM-CSF, which, in turn, is required for allergen induced pulmonary eosinophilia, identifying a novel pathway of eosinophil associated effector functions leading to pulmonary pathology in diseases such as asthma. PMID- 11907118 TI - Tissue-specific mechanisms control the retention of IL-8 in lungs and skin. AB - Chemokines are a group of structurally related peptides that promote the directed migration of leukocytes in tissue. Mechanisms controlling the retention of chemokines in tissue are not well understood. In this study we present evidence that two different mechanisms control the persistence of the CXC chemokine, IL-8, in lungs and skin. (125)I-labeled IL-8 was injected into the airspaces of the lungs and the dermis of the skin and the amount of (125)I-labeled IL-8 that remained at specified times was measured by scintillation counting. The (125)I labeled IL-8 was cleared much more rapidly from skin than lungs, as only 2% of the (125)I-labeled IL-8 remained in skin at 4 h whereas 50% of the (125)I-labeled IL-8 remained in lungs at 4 h. Studies in neutropenic rabbits showed that neutrophils shortened the retention of (125)I-labeled IL-8 in skin but not lungs. A monomeric form of IL-8, N-methyl-leucine 25 IL-8, was not retained as long in lungs as recombinant human IL-8, indicating that dimerization of IL-8 is a mechanism that increases the local concentration and prolongs the retention of (125)I-labeled IL-8 in lungs. These observations show that the mechanisms that control the retention of IL-8 in tissue include neutrophil migration and dimerization, and that the importance of these varies in different tissues. PMID- 11907119 TI - Analysis of the gene expression profile activated by the CC chemokine ligand 5/RANTES and by lipopolysaccharide in human monocytes. AB - The gene expression profile induced by the CC chemokine ligand (CCL) 5/RANTES in human monocytes was examined using the oligonucleotide array technology. Of 5600 transcripts examined, 42 were consistently induced by CCL5, and none were suppressed. Chemokine-inducible transcripts could be clustered in functional groups, including selected cytokines and receptors (e.g., IL-1beta, CCL2/monocyte chemotactic protein-1, and the CCL5 receptor CCR1) and molecules involved in extracellular matrix recognition and digestion (e.g., CD44 splice transcripts, urokinase-type plasminogen activator receptor, matrix metalloprotease (MMP)-9, and MMP-19). Transcript expression, confirmed by quantitative real-time PCR analysis for selected genes, was associated with protein induction for some (e.g., CCL2), but not all (e.g., IL-1beta), transcripts examined. The chemokine induced gene profile was distinct from that activated by LPS, a prototypic phagocyte activator. Although certain transcripts were stimulated by both agonists (e.g., IL-1beta and CCL2), others were induced only by either LPS (e.g., TNF-alpha and IL-6) or CCL5 (e.g., MMP-19) or were divergently regulated (e.g., CCR1). Thus, CCL5, a prototypic CC inflammatory chemokine, activates a restricted transcriptional program in monocytes distinct from that induced by the prototypic pathogen-derived proinflammatory stimulant LPS. Chemokine-induced chemokines production could represent a novel amplification loop of leukocyte recruitment, while a subset of chemokine-inducible transcripts could be involved in monocyte extravasation and tissue invasion. PMID- 11907120 TI - 15-Deoxy-delta 12,14-prostaglandins D2 and J2 are potent activators of human eosinophils. AB - 15-Deoxy-Delta(12,14)-PDJ(2) (15d-PGJ(2)) is a degradation product of PGD(2) that has been proposed as an anti-inflammatory compound because of its various inhibitory effects, some of which are mediated by peroxisome proliferator activated receptor-gamma. In contrast to its reported inhibitory effects on macrophages and other cells, we found that this compound is a potent activator of eosinophils, inducing calcium mobilization, actin polymerization, and CD11b expression. It is selective for eosinophils, having little or no effect on neutrophils or monocytes. 15d-PGJ(2) has an EC(50) of approximately 10 nM, similar to that of its precursor, PGD(2). The concentrations of 15d-PGJ(2) required to activate eosinophils are thus much lower than those required for its anti-inflammatory effects (usually micromolar). 15-Deoxy-Delta(12,14) prostaglandin D(2) (15d-PGD(2)) is also a potent activator of eosinophils, with an EC(50) about the same as that of PGD(2), whereas Delta(12)-PGJ(2) is slightly less potent. Eosinophils pretreated with PGD(2) no longer respond to 15d-PGJ(2), and vice versa, but in both cases the cells still respond to another eicosanoid proinflammatory mediator, 5-oxo-6,8,11,14-eicosatetraenoic acid. This indicates that the effects of 15d-PGJ(2) are mediated by the DP(2)/chemoattractant receptor homologous molecule expressed on Th2 cells that has recently been identified in eosinophils. 15d-PGJ(2) is selective for the DP(2) receptor, in that it has no effect on DP(1) receptor-mediated adenylyl cyclase activity in platelets. We conclude that 15d-PGJ(2) and 15d-PGD(2) are selective DP(2) receptor agonists that activate human eosinophils with potencies at least 100 times greater than those for the proposed anti-inflammatory effects of 15d-PGJ(2) on other cells. PMID- 11907121 TI - Modulation of leukotriene B4 receptor-1 expression by dexamethasone: potential mechanism for enhanced neutrophil survival. AB - Glucocorticoids can down-regulate many inflammatory and immune responses and constitute a powerful therapeutic tool in a number of diseases. However, they have a somewhat paradoxical effect on neutrophils, in that they prolong their survival. Because leukotriene B(4) (LTB(4)) can also extend neutrophil survival, we proposed that glucocorticoids could prevent neutrophil apoptosis by up regulating their expression of the high-affinity LTB(4) receptor (BLT1). Here we show that, indeed, dexamethasone (DEX) up-regulates the steady-state levels of BLT1 mRNA in human neutrophils. The effect was time and concentration dependent, being maximal at 4 h and at 10-100 nM DEX. The effect was also dependent on transcriptional activity, whereas BLT1 mRNA stability was not affected. DEX induced up-regulation of BLT1 expression was prevented by pretreatment with the LTB(4) antagonist LY255283. Moreover, LTB(4) itself up-regulated the expression of BLT1 mRNA. BLT1 protein expression on neutrophils exposed to DEX for 24 h was also up-regulated 2- to 3-fold, and DEX-treated as well as LTB(4)-treated cells showed enhanced responsiveness to LTB(4) in terms of intracellular Ca(2+) mobilization and chemotaxis. Whereas DEX and LTB(4) alone decreased neutrophil apoptosis by approximately 50%, neutrophils treated with both LTB(4) and DEX showed >90% survival at 24 h. Moreover, BLT1 antagonists prevented the increased neutrophil survival induced by DEX as well as by LTB(4). Taken together, our results suggest that DEX-induced up-regulation of BLT1 expression in neutrophils may be one mechanism through which glucocorticoids can prolong neutrophil survival, namely by enhancing cell responses to the antiapoptotic effect of LTB(4). PMID- 11907122 TI - Activation of protease-activated receptor (PAR)-1, PAR-2, and PAR-4 stimulates IL 6, IL-8, and prostaglandin E2 release from human respiratory epithelial cells. AB - Epithelia from many tissues express protease-activated receptors (PARs) that play a major role in several different physiological processes. In this study, we examined their capacity to modulate IL-6, IL-8, and PGE(2) production in both the A459 and BEAS-2B cell lines and primary human bronchial epithelial cells (HBECs). All three cell types expressed PAR-1, PAR-2, PAR-3, and PAR-4, as judged by RT PCR and immunocytochemistry. Agonist peptides corresponding to the nascent N termini of PAR-1, PAR-2, and PAR-4 induced the release of cytokines from A549, BEAS-2B, and HBECs with a rank order of potency of PAR-2 > PAR-4 > PAR-1 at 400 microM. PAR-1, PAR-2, and PAR-4 also caused the release of PGE(2) from A549 and HBECs. The PAR-3 agonist peptide was inactive in all systems tested. PAR-1, PAR 2, or PAR-4, in combination, caused additive IL-6 release, but only the PAR-1 and PAR-2 combination resulted in an additive IL-8 response. PAR peptide-induced responses were accompanied by changes in intracellular calcium ion concentrations. However, Ca(2+) ion shutoff was approximately 2-fold slower with PAR-4 than with PAR-1 or PAR-2, suggesting differential G protein coupling. Combined, these data suggest an important role for PAR in the modulation of inflammation in the lung. PMID- 11907123 TI - Distinct temporal patterns of macrophage-inflammatory protein-2 and KC chemokine gene expression in surgical injury. AB - In the present study the regulation of CXC chemokine expression was evaluated in full-thickness abdominal wounds in mice. During the first 24 h after injury, IL 1alphabeta, KC, macrophage-inflammatory protein (MIP)-2, and monocyte chemoattractant protein-1 were the predominant cytokines and chemokines produced; TNF-alpha was not detected. Chemokine mRNA expression and protein secretion occurred in two temporal stages. The first, which reached a maximum at 6 h, was associated with high levels of IL-1alpha and KC and low levels of MIP-2. This stage could be reproduced by intradermal injection of IL-1alpha or IL-1beta and was partially blocked by injection of neutralizing Ab against IL-1alpha but not IL-1beta. In animals depleted of circulating neutrophils, chemokine expression was reduced by nearly 70% during this stage. In the second stage, which peaked at 24 h after injury, modest but significant levels of IL-1beta were detected in association with low levels of KC and high levels of MIP-2. This pattern of chemokine expression could not be mimicked by injection of IL-1alpha or IL-1beta (even with prolonged exposure), although MIP-2 expression could be partially inhibited by intradermal injection of neutralizing Ab against IL-1beta. Surprisingly, neutrophil depletion before injury resulted in sustained high levels of both KC and MIP-2 expression. These observations demonstrate that these two closely related chemokines are under distinct regulatory controls in vivo that are likely to reflect the temporally ordered participation of different cell types and/or extracellular stimuli and inhibitors. PMID- 11907124 TI - Contribution of vascular endothelial growth factor to airway hyperresponsiveness and inflammation in a murine model of toluene diisocyanate-induced asthma. AB - Isocyanate chemicals, including toluene diisocyanate (TDI), are currently the most common causes of occupational asthma. Although considerable controversy remains regarding its pathogenesis, TDI-induced asthma is characterized by hyperresponsiveness and inflammation of the airways. One of the histological hallmarks of inflammation is angiogenesis, but the possible role of vascular endothelial growth factor (VEGF), a potent angiogenic cytokine, in TDI-induced asthma is unknown. We developed a murine model to investigate TDI-induced asthma by performing two courses of sensitization with 3% TDI and one challenge with 1% TDI using ultrasonic nebulization to examine the potential involvement of VEGF in that disease. These mice develop the following typical pathophysiological features: airway hyperresponsiveness, airway inflammation, and increased VEGF levels in the airway. Administration of VEGFR inhibitors reduced all these pathophysiological symptoms. These results suggest that VEGF is one of the major determinants of TDI-induced asthma and that the inhibition of VEGF may be a good therapeutic strategy. PMID- 11907125 TI - Transgenic expression of a soluble complement inhibitor protects against renal disease and promotes survival in MRL/lpr mice. AB - To investigate the role of complement in lupus nephritis, we used MRL/lpr mice and a transgene overexpressing a soluble complement regulator, soluble CR1 related gene/protein y (sCrry), both systemically and in kidney. Production of sCrry in sera led to significant complement inhibition in Crry-transgenic mice relative to littermate transgene negative controls. This complement inhibition with sCrry conferred a survival advantage to MRL/lpr mice. In a total of 154 animals, 42.5% transgene-negative animals had impaired renal function (blood urea nitrogen > 50 mg/dl) compared with 16.4% mice with the sCrry-producing transgene (p < 0.001). In those animals that died spontaneously, MRL/lpr mice with the sCrry-producing transgene did not die of renal failure, while those without the transgene did (blood urea nitrogen values of 46.6 +/- 9 and 122 +/- 29 mg/dl in transgene-positive and transgene-negative animals, respectively; p < 0.001). Albuminuria was reduced in those transgenic animals in which sCrry expression was maximally stimulated (urinary albumin/creatinine = 12.4 +/- 4.3 and 36.9 +/- 7.7 in transgene-positive and transgene-negative animals, respectively; p < 0.001). As expected in the setting of chronic complement inhibition, there was less C3 deposition in glomeruli of sCrry-producing transgenic mice compared with transgene-negative animals. In contrast, there was no effect on glomerular IgG deposition, levels of anti-dsDNA Ab and rheumatoid factor, or spleen weights between the two groups. Thus, long-term complement inhibition reduces renal disease in MRL/lpr mice, which translates into improved survival. MRL/lpr mice in which complement is inhibited still have spontaneous mortality, yet this is not from renal disease. PMID- 11907126 TI - IL-18-binding protein expression by endothelial cells and macrophages is up regulated during active Crohn's disease. AB - The pathogenesis of Crohn's disease (CD) remains under intense investigation. Increasing evidence suggests a role for mature IL-18 in the induction of proinflammatory cytokines and Th1 polarization in CD lesions. The aim of this study was to investigate the contribution of the IL-18-neutralizing (a and c) and non-neutralizing (b and d) isoforms of IL-18-binding protein (IL-18BP) during active CD. Intestinal endothelial cells and macrophages were the major source of IL-18BP within the submucosa, and this IL-18BP production was also found to be relevant to other types of endothelial cells (HUVEC) and macrophages (peripheral monocytes). IL-18BP messenger transcript and protein were significantly increased in surgically resected specimens from active CD compared with control patients, correlating with an up-regulation of IL-18. Analysis of the expression of the four IL-18BP isoforms as well as being free or bound to IL-18 was reported and revealed that unbound IL-18BP isoforms a and c and inactive isoform d were present in specimens from active CD and control patients while isoform b was not detected. IL-18/IL-18BP complex was also detected. Interestingly, although most was complexed, free mature IL-18 could still be detected in active CD specimens even in the presence of the IL-18BP isoform a/c. These results demonstrate that the appropriate neutralizing isoforms are present in the intestinal tissue of patients with active CD and highlights the complexity of IL-18/IL-18BP biology. PMID- 11907127 TI - A peptide DNA surrogate accelerates autoimmune manifestations and nephritis in lupus-prone mice. AB - Lupus-associated anti-DNA Abs display features of Ag selection, yet the triggering Ag in the disease is unknown. We previously demonstrated that the peptide DWEYSVWLSN is bound by a pathogenic anti-DNA Ab, and that immunization of nonautoimmune mice with this peptide induces autoantibodies and renal Ig deposition. To elucidate differences in the induced B cell responses in mice genetically predisposed to autoimmunity, young (NZB x NZW)F(1) mice were immunized with this peptide DNA mimetope. DWEYSVWLSN-immunized mice had significantly increased IgG anti-dsDNA, anti-laminin, and anti-cardiolipin Ab titers compared with controls. In addition, glomerular histopathology in the form of endocapillary disease and crescent formation was markedly more severe in DWEYSVWLSN-immunized mice. Analysis of mAbs from DWEYSVWLSN-immunized mice revealed that anti-peptide Abs were often cross-reactive with DNA. Genetic elements used in the Ab response in immunized mice were homologous to those used in the spontaneous anti-DNA response in (NZB x NZW)F(1) mice, as well as in other, experimentally induced anti-DNA Abs. Our results indicate that peptide immunization can induce a molecular genetic response common to a variety of stimuli that break tolerance to mammalian dsDNA. Based on the similarity between spontaneously arising anti-DNA Abs and several types of induced anti-DNA Abs, we suggest that there may be more than a single Ag that can trigger systemic lupus erythematosus. PMID- 11907128 TI - Beryllium skin patch testing to analyze T cell stimulation and granulomatous inflammation in the lung. AB - Chronic beryllium disease (CBD) is characterized by granulomatous inflammation and the accumulation of CD4(+) T cells in the lung. Patch testing of CBD patients with beryllium sulfate results in granulomatous inflammation in the skin. We investigated whether the T cell clonal populations present in the lung of CBD patients would also be present in the involved skin of a positive beryllium patch test and thus mirror the granulomatous process in the lung. CBD patients with clonal TCR expansions in bronchoalveolar lavage (BAL) were selected for study. All three CBD patients studied had a positive response to beryllium sulfate application and a negative patch test to normal saline. Immunohistochemistry showed extensive infiltration with CD4(+) T cells and few, if any, CD8(+) T cells both at 3 days and at later times when granulomas were apparent. T cell infiltration early after skin testing appeared to be nonspecific with the TCR repertoire of infiltrating T cells being distinct from that present in BAL. At later times when granulomas were present, T cell clones in skin overlapped with those in BAL in all patients tested. Total TCR matches in skin and BAL were as high as 40% in selected Vbeta T cell subsets. Studies of peripheral blood T cells before and after patch testing provided evidence for mobilization of large numbers of pathogenic beryllium-reactive T cells into the circulating pool. These studies using skin patch testing provide new insight into the dynamics of T cell influx and mobilization during granulomatous inflammation. PMID- 11907129 TI - Human DQ8 can substitute for murine I-Ag7 in the selection of diabetogenic T cells restricted to I-Ag7. AB - The strong association of type 1 diabetes with specific MHC class II genes, such as I-A(g7) in nonobese diabetic mice and HLA-DQ8 in humans, suggests that MHC class II molecules play an important role in the development of the disease. To test whether human DQ8 molecules could cross the species barrier and functionally replace their murine homolog I-A(g7), we generated DQ8/BDC2.5 transgenic mice. We have shown that BDC2.5 transgenic T cells are selected on DQ8 in the thymus and cause diabetes in a manner similar to that seen when the T cells are selected on H2(g7). Splenocytes from DQ8/BDC2.5 mice also showed reactivity toward islets in vitro as seen in H-2(g7)/BDC2.5 mice. We conclude that DQ8 molecules not only share structural similarity with the murine homolog I-A(g7), but also can cross the species barrier and functionally replace I-A(g7) molecules to stimulate diabetogenic T cells and produce diabetes. PMID- 11907130 TI - Long-term reversal of established autoimmunity upon transient blockade of the LFA 1/intercellular adhesion molecule-1 pathway. AB - Transgenic models and administration of mAbs directed against the LFA 1/intercellular adhesion molecule 1 (ICAM-1) pathway have shown that these costimulatory molecules play a key role in generating effector cells mediating inflammatory responses. In this report, durable remission of recent diabetes in nonobese diabetic (NOD) mice was induced by transient expression of an immunoadhesin gene encoding the soluble form of ICAM-1 (sICAM-1/Ig). A single i.v. injection of an adenovirus vector encoding the immunoadhesin gene led to 70% diabetes remission as opposed to 0% in mice injected with a control adenovirus vector. Despite the rapid decline of sICAM-1/Ig serum levels, diabetes remission remained stable in 50% of NOD mice for >6 mo. sICAM-1/Ig expression also led to long-term protection against diabetes in prediabetic NOD mice. sICAM-1/Ig in vitro induced an agonistic effect of T cell activation in a TCR-transgenic model, increasing T cell proliferation and IL-2 secretion. Importantly, protected mice were not immunosuppressed because they rejected skin allografts normally and developed immunity against the adenovirus vector. Rather, sICAM-1/Ig induced active tolerance, as assessed by the persistence of diabetogenic T cells in protected mice and the reversal of protection by immunosuppression with cyclophosphamide. PMID- 11907131 TI - Oligoclonal T cell expansion in the skin of patients with systemic sclerosis. AB - Fibrosis, microvascular fibroproliferative alterations, and autoantibody production are the main features of systemic sclerosis (SSc), and all of them can be explained by cytokine production by activated T cells. However, little is known about the role of T cells in the pathogenesis of SSc, and there is no information on the Ag(s) that elicits such activation. To determine whether T cells infiltrating the skin biopsies of patients with SSc are oligoclonal, beta chain TCR transcripts from T cells infiltrating the skin of five patients with SSc of recent onset were amplified by either Vbeta-specific PCR or nonpalindromic adaptor PCR. The resulting PCR products were subsequently cloned and sequenced. High proportions of identical beta-chain TCR transcripts ranging from 43 to 90% of those sequenced were found in five patients, strongly suggesting the presence of oligoclonal T cells in these infiltrates. A dominant T cell clone was found to be clonally expanded in skin biopsies obtained from a single patient with SSc at three different times (0, 8, and 13 mo earlier) and from three different skin regions. beta-chain TCR transcripts from PBMC from normal donors (methodological control) were unique when compared with each other, typical for polyclonal populations of T cells. The finding of oligoclonal T cells infiltrating the skin of patients with SSc suggests that these T cells have undergone proliferation in situ in the skin and clonal expansion in response to as yet unidentified Ag(s). These results suggest that T cells are involved in the pathogenesis of the disease. PMID- 11907132 TI - Dynamics of T cell responses in HIV infection. AB - Cytotoxic CD8(+) T cells play a major role in the immune response against viruses. However, the dynamics of CD8(+) T cell responses during the course of a human infection are not well understood. Using tetrameric complexes in combination with a range of intracellular and extracellular markers, we present a detailed analysis of the changes in activation and differentiation undergone by Ag-specific CD8(+) T cells, in relation to Ag-specific CD4(+) T cell responses, in the context of a human infection: HIV-1. During primary HIV-1 infection, the initial population of HIV-specific CD8(+) T cells is highly activated and prone to apoptosis. The Ag-specific cells differentiate rapidly from naive to cells at a perforin low intermediate stage of differentiation, later forming a stable pool of resting cells as viral load decreases during chronic infection. These observations have significant implications for our understanding of T cell responses in human viral infections in general and indicate that the definition of effector and memory subsets in humans may need revision. PMID- 11907133 TI - Antibodies against cerebral M1 cholinergic muscarinic receptor from schizophrenic patients: molecular interaction. AB - We demonstrated the presence of circulating Abs from schizophrenic patients able to interact with cerebral frontal cortex-activating muscarinic acetylcholine receptors (mAChR). Sera and purified IgG from 21 paranoid schizophrenic and 25 age-matched normal subjects were studied by indirect immunofluorescence, flow cytometry, immunoblotting, dot blot, ELISA, and radioligand competition assays. Rat cerebral frontal cortex membranes and/or a synthetic peptide, with an amino acid sequence identical with that of human M(1) mAChR, were used as Ags. By indirect immunofluorescence and flow cytometry procedures, we proved that serum purified IgG fraction from schizophrenic patients reacted to neural cell surfaces from rat cerebral frontal cortex. The same Abs were able to inhibit the binding of the specific M(1) mAChR radioligand [(3)H]pirenzepine. Immunoblotting experiments showed that IgG from schizophrenic patients revealed a band with a molecular mass coincident to that labeled by an anti-M(1) mAChR Ab. Using synthetic peptide for dot blot and ELISA, we demonstrated that these Abs reacted against the second extracellular loop of human cerebral M(1) mAChR. Also, the corresponding affinity-purified antipeptide Ab displayed an agonistic-like activity associated to specific receptor activation, increasing cyclic GMP production and inositol phosphate accumulation, and protein kinase C translocation. This paper gave support to the participation of an autoimmune process in schizophrenia. PMID- 11907134 TI - Spleen is a primary site for activation of platelet-reactive T and B cells in patients with immune thrombocytopenic purpura. AB - We have recently reported that in patients with chronic immune thrombocytopenic purpura (IMTP), circulating T and B cells that are responsive to gpIIb-IIIa can induce anti-platelet autoantibody production. In this study, the frequencies and activation status of gpIIb-IIIa-reactive T and B cells were evaluated in the peripheral blood and spleen obtained from nine IMTP patients undergoing splenectomy. There was no difference in gpIIb-IIIa-reactive T cell frequencies between peripheral blood and spleen (6.4 +/- 2.6 vs 5.2 +/- 2.4 per 10(5) T cells), as determined by limiting dilution analysis, but activated T cells responsive to gpIIb-IIIa showing accelerated proliferation kinetics and those expressing CD154 were more frequent in spleen than in peripheral blood. The frequencies of anti-gpIIb-IIIa Ab-producing B cells, as determined by ELISPOT assay, were also similar in peripheral blood and spleen (61.2 +/- 24.0 vs. 77.7 +/- 45.3 per 10(5) B cells); however, an anti-gpIIb-IIIa Ab was spontaneously produced by splenocytes in vitro, but scarcely secreted by PBMCs. CD19(-)/surface Ig(-)/CD38(+)/CD138(+) plasma cells secreting anti-gpIIb-IIIa Ab were exclusively detected in the spleen. In serial analysis, the frequencies of circulating gpIIb IIIa-reactive T and B cells were markedly decreased after splenectomy in patients with a complete response, but were unchanged in nonresponders. These findings indicate that an interaction between gpIIb-IIIa-reactive T and B cells inducing anti-platelet Ab production in IMTP patients occurs primarily in the spleen and that the significant number of gpIIb-IIIa-reactive T and B cells activated in the spleen are released into the circulation as memory cells. PMID- 11907135 TI - Regulation of cholesterol-7alpha-hydroxylase: BAREly missing a SHP. AB - Cholesterol-7alpha-hydroxylase (CYP7A1) regulates the pathway through which cholesterol is converted into bile acids. The unique detergent properties of bile acids are essential for the digestion and intestinal absorption of hydrophobic nutrients. Bile acids have potent toxic properties (e.g., membrane disruption) and there are a plethora of mechanisms to limit their accumulation in blood and tissues. The discovery of farnesoid X receptor (FXR), the nuclear receptor activated specifically by bile acids, has opened new insights into these mechanisms. Bile acid activation of FXR has been shown to repress the expression of CYP7A1 via increasing the expression of small heterodimer partner (SHP), a non DNA binding protein. The increased abundance of SHP causes it to associate with liver receptor homolog (LRH)-1, an obligate factor required for transcription of CYP7A1. Recent studies show there is an "FXR/SHP-independent" mechanism that also represses CYP7A1 expression. This "FXR/SHP-independent" pathway involves the interaction of bile acids with liver macrophages (i.e., Kupffer cells), which induces the expression, and secretion of cytokines. These inflammatory cytokines, which include tumor necrosis factor alpha and interleukin-1beta, act upon liver parenchymal cells causing a rapid repression of the CYP7A1 gene. PMID- 11907136 TI - SR-BI is required for microvillar channel formation and the localization of HDL particles to the surface of adrenocortical cells in vivo. AB - Scavenger receptor class B type I (SR-BI) mediates the selective uptake of HDL cholesteryl ester into liver and steroidogenic tissues. In steroidogenic cells, juxtaposed microvilli, or microvilli snuggled against the plasma membrane create microvillar channels that fill with HDL. Microvillar membranes contain SR-BI and are believed to be the site of HDL cholesteryl ester uptake. A recent study showed that SR-BI expression in insect cells elicits membrane structures that contain SR-BI, bind HDL, and closely resemble the ultrastructure of microvillar channels. In the present study we compared the ultrastructure of adrenal gland microvillar membranes in Srb1+/+ and Srb1-/- mice to test whether SR-BI is required for the formation of microvillar channels. The results show that SR-BI is absolutely required for microvillar channel formation and that the microvillar membranes of Srb1-/- mice are 17% thinner than in Srb1+/+ mice. We conclude that SR-BI has a major influence on plasma membrane ultrastructure and organization in vivo. PMID- 11907137 TI - Prompt inhibition of fMLP-induced Ca2+ mobilization by parenteral lipid emulsions in human neutrophils. AB - It remains unclear whether modulation of immune system functions by lipids contributes to the increased infection rate observed in patients treated with parenteral nutrition. We therefore evaluated the effects of lipid emulsions derived from fish oil [very long chain triglycerides (VLCT)], olive oil [long chain triglycerides- mono-unsaturated fatty acid (LCT-MUFA)], soya oil [long chain triglycerides (LCT)], or a physical mixture of coconut and soya oil [mixed long- and medium-chain triglycerides (LCT-MCT)] on neutrophil activation. N formyl-methionyl-leucyl-phenylalanine (fMLP) evoked an immediate increase of the cytosolic Ca2+ concentration ([Ca2+](i,av)) in a suspension of neutrophils. When added 3 min before fMLP, however, all four lipid emulsions reduced the hormone induced increase in [Ca2+](i,av) with the same efficacy but with different potency. Half-maximal inhibition was reached at emulsion concentrations of 0.24 mM VLCT, 0.32 mM LCT-MCT, 0.52 mM LCT, and 0.82 mM LCT-MUFA. Similarly to the lipids, the protein kinase C (PKC) activator PMA markedly reduced the fMLP induced increase in [Ca2+](i,av). PMA inhibition was abolished by the PKC inhibitor staurosporine. In contrast, however, this drug did not interfere with the inhibitory lipid effect, indicating that the lipids act primarily in a PKC independent manner. In summary, this study shows that nutritional lipids can evoke a prompt and significant attenuation of hormone-induced neutrophil stimulation and that the emulsions based on fish oil and a mixture of coconut oil and soya oil are among the most potent ones in this respect. PMID- 11907138 TI - Dietary oxidized fatty acids may enhance intestinal apolipoprotein A-I production. AB - Apolipoprotein (apo)A-I, the major protein component of HDL, is synthesized principally in the small intestine and liver. Recently we observed an increase in plasma apoA-I level in humans who were on an oxidized fat diet. To test whether oxidized fatty acids could affect apoA-I synthesis, we incubated day 4 (undifferentiated) and day 14 (differentiated) Caco-2 cells with varying concentrations of oxidized linoleic acid (ox-linoleic acid) (5, 10, and 25 microM) and unoxidized linoleic acid for 24 h. Ox-linoleic acid caused a dose dependent increase in the levels of apoA-I protein in both differentiated and undifferentiated Caco-2 cells as assessed by ELISA and Western blot analysis. Whereas apoB production was not increased by ox-linoleic acid in both day 4 and day 14 Caco-2 cells. The mRNA expression for apoA-I paralleled the protein expression when measured by RT-PCR. We also found that both day 4 and day 14 Caco 2 cells did express peroxisomal proliferator-activated receptor-gamma (PPAR gamma). mRNA and PPAR-gamma ligand could increase apoA-I secretion in these cells. Therefore we propose that the mechanism for the induction of apoA-I might include PPAR-gamma for which oxidized fatty acid is a ligand. PMID- 11907139 TI - Molecular cloning, genomic organization, genetic variations, and characterization of murine sterolin genes Abcg5 and Abcg8. AB - Mammalian physiological processes can distinguish between dietary cholesterol and non-cholesterol, retaining very little of the non-cholesterol in their bodies. We have recently identified two genes, ABCG5 and ABCG8, encoding sterolin-1 and -2 respectively, mutations of which cause the human disease sitosterolemia. We report here the mouse cDNAs and genomic organization of Abcg5 and Abcg8. Both genes are arranged in an unusual head-to-head configuration, and only 140 bases separate their two respective start-transcription sites. A single TATA motif was identified, with no canonical CCAT box present between the two genes. The genes are located on mouse chromosome 17 and this complex spans no more than 40 kb. Expression of both genes is confined to the liver and intestine. For both genes, two different sizes of transcripts were identified which differ in the lengths of their 3' UTRs. Additionally, alternatively spliced forms for Abcg8 were identified, resulting from a CAG repeat at the intron 1 splice-acceptor site, causing a deletion of a glutamine. We screened 20 different mouse strains for polymorphic variants. Although a large number of polymorphic variants were identified, strains reported to show significant differences in cholesterol absorption rates did not show significant genomic variations in Abcg5 or Abcg8. PMID- 11907140 TI - Niemann-Pick C1 protein regulates cholesterol transport to the trans-Golgi network and plasma membrane caveolae. AB - The Niemann-Pick C1 (NPC1) protein regulates cholesterol transport from late endosomes-lysosomes to other intracellular compartments. In this article, cholesterol transport to caveolin-1 and caveolin-2 containing compartments, such as the trans-Golgi network (TGN) and plasma membrane caveolae, was examined in normal (NPC+/+), NPC heterozygous (NPC+/-), and NPC homozygous (NPC-/-) human fibroblasts. The expression and distribution of NPC1 in each cell type were similar, and characterized by a finely dispersed, granular staining pattern. The expression of caveolin-1 and caveolin-2 was increased in NPC+/- and NPC-/- fibroblasts, although the distribution in each cell type was similar and characterized by predominant staining of the TGN and plasma membrane. The TGN in NPC+/+ fibroblasts was relatively cholesterol-enriched, whereas the TGN in NPC+/- and NPC-/- fibroblasts was partially or completely cholesterol-deficient, respectively. Consistent with studies demonstrating the transport of cholesterol from the TGN to plasma membrane caveolae, the concentration of cholesterol in plasma membrane caveolae isolated from NPC+/- and NPC-/- fibroblasts was significantly decreased, even though the total concentration of plasma membrane cholesterol in each cell type was similar. These studies demonstrate that NPC1 regulates cholesterol transport to caveolin-1 and caveolin-2 containing compartments such as the TGN and plasma membrane caveolae. PMID- 11907141 TI - Targeted disruption of the mouse cis-retinol dehydrogenase gene: visual and nonvisual functions. AB - It has been proposed that cis-retinol dehydrogenase (cRDH) acts within the body to catalyze the oxidation of 9-cis-retinol, an oxidative step needed for 9-cis retinoic acid synthesis, the oxidation of 11-cis-retinol [an oxidative step needed for 11-cis-retinal (visual chromophore) synthesis], and 3 alpha hydroxysteroid transformations. To assess in vivo the physiological importance of each of these proposed actions of cRDH, we generated cRDH-deficient (cRDH-/-) mice. The cRDH-/- mice reproduce normally and appear to be normal. However, the mutant mice do have a mild visual phenotype of impaired dark adaptation. This phenotype is evidenced by electroretinagram analysis of the mice and by biochemical measures of eye levels of retinoid intermediates during recovery from an intense photobleach. Although it is thought that cRDH is expressed in the eye almost solely in retinal pigment epithelial cells, we detected cRDH expression in other retinal cells, including ganglion cells, amacrine cells, horizontal cells, and the inner segments of the rod photoreceptor cells. Aside from the eye, there are no marked differences in retinoid levels in other tissues throughout the body for cRDH-/- compared with cRDH+/+ mice. Moreover, we did not detect any non visual phenotypic changes for cRDH-/- mice, suggesting that these mice do not have problems in metabolizing 3 alpha-hydroxysteroids.Thus, cRDH may act essentially in the visual cycle but is redundant for catalyzing 9-cis-retinoic acid formation and 3 alpha-hydroxysteroid metabolism. PMID- 11907142 TI - LDL particle size in familial combined hyperlipidemia: effects of serum lipids, lipoprotein-modifying enzymes, and lipid transfer proteins. AB - Small, dense LDL particles are typical for FCHL. Intravascular lipid exchange and net transfer among HDL, LDL, and triglyceride-rich lipoproteins as well as lipolysis in the VLDL-IDL-LDL cascade regulate properties of LDL. We investigated postheparin plasma activities of hepatic lipase (HL) and LPL, and plasma activities of CETP and phospholipid transfer protein (PLTP) in 191 individuals from 37 Finnish FCHL families. LDL peak particle diameter (LDL size) was measured with 2-10% gradient polyacrylamide gel electrophoresis. LDL size was significantly smaller in affected FCHL family members (n = 68) as compared with nonaffected FCHL family members (n = 78) or spouses (n = 45) (25.3 +/- 1.5 nm, 26.8 +/- 1.2 nm, and 26.6 +/- 1.2 nm, respectively, P < 0.001 for both). In affected FCHL family members, serum triglycerides were the strongest correlate for LDL size (r = -0.71, P < 0.001). In univariate correlation analysis LDL size was not associated with HL, LPL, CETP, and PLTP activities. In multivariate stepwise regression analysis, however, serum triglycerides, CETP activity, HL activity, and HDL cholesterol were significant predictors of LDL size in affected FCHL subjects (adjusted r (2) = 0.642). We conclude that serum triglyceride concentration is strongly correlated with LDL size in affected FCHL subjects. After adjustment for serum triglycerides, HL and CETP activities are associated with LDL size in FCHL. PMID- 11907143 TI - Quantitation of cholesterol crystallization from supersaturated model bile. AB - Cholesterol crystallization is an essential step in gallstone formation. Although spectrophotometry and nephelometry have been used for quantitation of crystallization, potential effects of crystal size and shape have not been evaluated. We determined crystallization in model biles [total lipid concentration 7.3 g/dl, egg yolk Phosphatidylcholine (EYPC)/(EYPC+taurocholate) molar ratio = 0.05, 0.15, or 0.30; cholesterol saturation index (CSI) = 1.2, 1.7, or 2.0; 37 degrees C] plotting in the central three-phase (micelles, vesicles, and crystals containing) zone or in the left two-phase (micelles and crystals containing) zone of the equilibrium ternary phase diagram. Extent of crystallization estimated by spectrophotometry and nephelometry was related to chemical determination of crystal mass and to crystal size or shape (by microscopy). With all methods, crystallization was less extensive when vesicles were present (central three-phase zone) and at lower CSIs. In the left two-phase zone, particularly at EYPC/(EYPC+taurocholate), ratio of 0.15, there were strong increases in spectrophotometric and nephelometric readings during the first days of incubation, but decreases at later stages, despite progressive increases in crystal mass by chemical measurement. Initially, there were large numbers of very small crystals (<10 microm) in these biles, which were subsequently replaced by large cholesterol monohydrate crystals. Decreasing sizes of harvested cholesterol monohydrate crystals by sonication increased spectrophotometric and nephelometric values despite identical crystal mass. When cholesterol crystal mass is assayed by indirect methods such as spectrophotometry or nephelometry, results are strongly influenced by crystal size. PMID- 11907144 TI - Differential effects of n-3 fatty acid deficiency on phospholipid molecular species composition in the rat hippocampus. AB - In this study, we have examined the effects of n-3 fatty acid deficient diets on the phospholipids (PL) molecular species composition in the hippocampus. Female rats were raised for two generations on diets containing linoleic acid (18:2n-6), with or without supplementation of alpha-linolenic acid (18:3n-3) or 18:3n-3 plus docosahexaenoic acid (22:6n-3). At 84 days of age, the hippocampal phospholipids were analyzed by reversed phase HPLC-electrospray ionization mass spectrometry. Depleting n-3 fatty acids from the diet led to a reduction of 22:6n-3 molecular species in phosphatidylcholine (PC), phosphatidylethanolamine (PE), PE plasmalogens (PLE), and phosphatidylserine (PS) by 70-80%. In general, 22:6n-3 was replaced with 22:5n-6 but the replacement at the molecular species level did not always occur in a reciprocal manner, especially in PC and PLE. In PC, the 16:0,22:6n-3 species was replaced by 16:0,22:5n-6 and 18:0,22:5n-6. In PLE, substantial increases of both 22:5n-6 and 22:4n-6 species compensated for the decreases in 22:6n-3 species in n-3 fatty acid deficient groups. While the total PL content was not affected by n-3 deficiency, the relative distribution of PS decreased by 28% with a concomitant increase in PC. The observed decrease of 22:6n-3 species along with PS reduction may represent key biochemical changes underlying losses in brain-hippocampal function associated with n-3 deficiency. PMID- 11907145 TI - Triglyceride depletion in THP-1 cells alters cholesteryl ester physical state and cholesterol efflux. AB - To study macrophage lipid droplet composition and the effects of TG on cholesteryl ester (CE) physical state, hydrolysis, and cholesterol efflux, a technique was developed to remove the majority of accumulated TG with minimal effect on CE content. THP-1 macrophages were incubated with acetylated LDL, and the accumulated TG was depleted by incubation with the acyl-CoA synthetase inhibitor triacsin D in the presence of albumin. Before TG removal, all cellular lipid droplets were isotropic as determined by polarizing light microscopy. When the TG concentration was reduced, anisotropic lipid droplets were visible, indicating a change in physical state, and suggesting that TG and CE originally accumulated in mixed lipid droplets. This change in physical state of lipid droplets was associated with slower rates of CE hydrolysis and cholesterol efflux. Although lipid droplets within the same cell had a similar physical state after TG depletion, there was considerable variability among cells in the physical state of their lipid droplets. In conclusion, THP-1 macrophages store accumulated CE and TG in mixed droplets, and the proportion of CE to TG varies among cells. Reducing accumulated TG altered CE physical state, which in turn affected hydrolysis of CE and cholesterol efflux. PMID- 11907146 TI - Expression of cholesterol-7alpha-hydroxylase in murine macrophages prevents cholesterol loading by acetyl-LDL. AB - Unlike macrophages, the hepatic parenchymal cells express cholesterol-7 alpha hydroxylase (CYP7A1) which regulates the conversion of cholesterol into bile acids, the major quantitative pathway maintaining cholesterol homeostasis. We examined if CYP7A1 expression in RAW 264.7 macrophages could prevent the accumulation of cholesterol when they were incubated with acetyl-LDL. A single cell clone (designated as 7 alphaRAW cells) that stably expresses rat CYP7A1 displayed similar rates of growth as non-transfected RAW cells. The CYP7A1 enzymatic activity expressed by microsomes obtained from 7 alphaRAW cells was similar to that expressed by microsomes obtained from mouse liver. Incubating non transfected RAW with increasing amounts of acetyl-LDL caused a parallel accumulation of cholesterol, whereas 7 alphaRAW cells displayed a complete resistance to cholesterol accumulation. 7 alphaRAW cells displayed increased expression of both ABCA1 mRNA (3.1-fold, P < 0.001) and ABCG1 mRNA (2.2-fold, P < 0.01), whereas the expression of scavenger receptor class A mRNA was unchanged. 7 alphaRAW cells also displayed small but significant increases in the rate of efflux of [(3)H]cholesterol to both delipidated apolipoprotein A1 and to HDL.Thus, CYP7A1 expression in RAW cultured macrophages prevented the accumulation of cholesterol from acetyl-LDL via both increased metabolism and efflux of cholesterol. PMID- 11907148 TI - Very long chain n-3 and n-6 polyunsaturated fatty acids bind strongly to liver fatty acid-binding protein. AB - Synthesis of n-3 and n-6 very long chain-PUFAs (VLC-PUFAs) from 18-carbon essential fatty acids is differentially regulated. The predominant product arising from n-3 fatty acids is docosahexaenoic acid (22:6n-3), with the liver serving as the main site of production. The synthetic pathway requires movement of a 24-carbon intermediate from the endoplasmic reticulum to peroxisomes for retroconversion to 22:6n-3. The mechanism of this intra-organelle flux is unknown, but could be binding-protein facilitated. We thus investigated binding of a series of previously untested VLC-PUFAs to liver fatty acid-binding protein (L-FABP). Three fluorometric assays were employed, all of which showed strong binding (K(d)' approximately 10(-8) to 10(-7) M) of 20-, 22-, and 24-carbon n-3 PUFAs to L-FABP. In contrast, synthesis of the predominant n-6 PUFA product, arachidonic acid, does not require intra-organelle transport. However, we found that n-6 VLC-PUFAs bound to L-FABP with affinities (K(d)' approximately 10(-8) to 10(-7) M) comparable to their n-3 counterparts. Although these results raise the possibility that L-FABP may participate in the cytoplasmic processing of n-3 and n-6 VLC-PUFAs, there is no evidence on the basis of binding affinities that L FABP accounts for differences in the predominant products formed by the n-3 and n 6 PUFA metabolic pathways. PMID- 11907147 TI - Molecular characterization of rabbit phospholipid transfer protein: choroid plexus and ependyma synthesize high levels of phospholipid transfer protein. AB - Phospholipid transfer protein (PLTP) plays an important role in plasma lipoprotein metabolism. However, PLTP is expressed in a wide range of tissues suggesting additional local functions. To analyze the tissue distribution of PLTP in an animal with high-level expression of the structurally and functionally related CETP, we have cloned the full-length cDNA of rabbit PLTP (1,796 bp). Rabbit PLTP cDNA shows high homology to human, murine, and porcine PLTP cDNA, averaging 86.1%, 80.4%, and 86.1%, respectively. Interestingly, the C-terminus contains a unique seven amino acid insertion not found in previously characterized mammalian PLTPs. In clear contradistinction to human PLTP, rabbit PLTP mRNA was prominent in brain. In situ hybridization studies revealed specific, high-level synthesis of PLTP mRNA in choroid plexus and ependyma, the organs responsible for production of cerebrospinal fluid. Consistent with these findings, PLTP activity in cerebrospinal fluid amounted to 23% +/- 3% of that in rabbit plasma. In contrast, neither CETP mRNA nor CETP activity were detectable in rabbit brain.A role of PLTP in the central nervous system could involve some of its actions previously established in vitro, like proteolysis of apolipoproteins, and be physiologically relevant for neurodegenerative disorders such as Alzheimer's disease. PMID- 11907149 TI - Separation of micelles and vesicles within lumenal aspirates from healthy humans: solubilization of cholesterol after a meal. AB - Understanding the physico-chemical relationship of lumenal lipids to one another is critical when elucidating the mechanism of components known to impact cholesterol absorption. Presently, there are no studies that describe the proportion of cholesterol carried as micelles or vesicles within human lumenal contents. Part of the reason for the scarceness of data is because of the lack of appropriate methodology required for reproducible sample collection and analysis. Thus, the object of the present studies was to develop a method to measure the amount of cholesterol carried as micelles or vesicles in human lumenal samples. The method includes the collection of lumenal samples from the ligament of Trietz through a Fredrick Miller tube, separation of the aqueous subphase from the nondigested lipids, separation of micelles and vesicles on Sepharose 4B columns within 48 h of collection using elution buffers consisting of the intermicellar bile acid composition, and finally quantitation of cholesterol eluted off of the columns. The distribution of cholesterol between micelles and vesicles obtained under different concentrations of bile acids and various lipids was comparable to results obtained from phase diagrams using the lumenal molar percentages of lipids obtained from the same samples. PMID- 11907150 TI - Pharmacology and toxicology of astrocyte-neuron glutamate transport and cycling. AB - The interaction between astrocytes and neurons is examined from the standpoint of glutamate neurotoxicity. The review details 1. the distribution of glutamate transporters on astrocytes and neurons, provoking a reformulation of the interdependence between these two cell types in removing extracellular glutamate and preventing excitotoxic injury; 2. the potential involvement of aberrant glutamate transporter function in the etiology of neuropathological conditions; 3. the role of astrocyte-neuron interaction in widely divergent aspects of brain energetics; 4. the role of astrocytes in the process of glutamate recycling within the context of anesthetic treatment with pentobarbital and thiopental. PMID- 11907151 TI - Role of multidrug transporters in pharmacoresistance to antiepileptic drugs. AB - Epilepsy, one of the most common neurologic disorders, is a major public health issue. Despite more than 20 approved antiepileptic drugs (AEDs), about 30% of patients are refractory to treatment. An important characteristic of pharmacoresistant epilepsy is that most patients with refractory epilepsy are resistant to several, if not all, AEDs, even though these drugs act by different mechanisms. This argues against epilepsy-induced alterations in specific drug targets as a major cause of pharmacoresistant epilepsy, but rather points to nonspecific and possibly adaptive mechanisms, such as decreased drug uptake into the brain by intrinsic or acquired over-expression of multidrug transporters in the blood-brain barrier (BBB). There is accumulating evidence demonstrating that multidrug transporters such as P-glycoprotein (PGP) and members of the multidrug resistance-associated protein (MRP) family are over-expressed in capillary endothelial cells and astrocytes in epileptogenic brain tissue surgically resected from patients with medically intractable epilepsy. PGP and MRPs in the BBB are thought to act as an active defense mechanism, restricting the penetration of lipophilic substances into the brain. A large variety of compounds, including many lipophilic drugs, are substrates for either PGP or MRPs or both. It is thus not astonishing that several AEDs, which have been made lipophilic to penetrate into the brain, seem to be substrates for multidrug transporters in the BBB. Over-expression of such transporters in epileptogenic tissue is thus likely to reduce the amount of drug that reaches the epileptic neurons, which would be a likely explanation for pharmacoresistance. PGP and MRPs can be blocked by specific inhibitors, which raises the option to use such inhibitors as adjunctive treatment for medically refractory epilepsy. However, although over-expression of multidrug transporters is a novel and reasonable hypothesis to explain multidrug resistance in epilepsy, further studies are needed to establish this concept. Furthermore, there are certainly other mechanisms of pharmacoresistance that need to be identified. PMID- 11907152 TI - Sustained activation of p38 mitogen-activated protein kinase contributes to the vascular response to injury. AB - The vascular response to mechanical injury involves inflammatory and fibroproliferative processes that result in the formation of neointima and vascular remodeling. The complex cellular interactions initiated by vascular injury are coordinated and modulated by the elaboration of cytokines and growth factors. The production and transduction of many of these mediators require phosphorylation of p38 mitogen-activated protein kinase (MAPK). In the present investigation, we examined the pattern and localization of p38 MAPK activation following balloon vascular injury. The effects of long-term and selective inhibition of p38 MAPK with SB 239063 (trans-1-(4-hydroxycyclohexyl)-4-(4 fluorophenyl)-5-[2-methoxy)pyrimidin-4-yl]imidazole) were also investigated in a model of vascular injury. Western blotting and immunohistochemical staining demonstrated that phospho-p38 MAPK was increased following balloon injury of the rabbit iliofemoral artery. The p38 MAPK activation was noted as early as 15 min after balloon injury and remained elevated for at least 28 days. Phospho-p38 MAPK immunoreactivity (IR) was localized primarily in regions of dedifferentiated, smooth muscle alpha-actin-positive cells in all lamina of the vessel wall. Phospho-p38 MAPK IR was not correlated with the localization of macrophage or proliferating cells (proliferating cell nuclear antigen; PCNA +). Long-term treatment (4 weeks) with SB 239063 (50 mg/kg/day, p.o.) reduced the vascular response to injury in the hypercholesterolemic rabbit. SB 239063 had no effect on platelet-derived growth factor (PDGF)-stimulated migration or proliferation of rabbit vascular smooth muscle cells (VSMCs) in culture. However, SB 239063 produced a concentration-dependent inhibition of transforming growth factor (TGF) beta-stimulated fibronectin production in VSMCs. In conclusion, sustained activation of p38 MAPK plays an important role in the vascular response to injury and inhibition of p38 MAPK may represent a novel therapeutic approach to limit this response. PMID- 11907153 TI - The angiotensin type 1 receptor antagonist, eprosartan, attenuates the progression of renal disease in spontaneously hypertensive stroke-prone rats with accelerated hypertension. AB - The effects of the angiotensin type 1 (AT(1)) receptor antagonist, eprosartan, were studied in a model of severe, chronic hypertension. Treatment of male spontaneously hypertensive stroke prone rats (SHR-SP) fed a high-fat, high-salt diet with eprosartan (60 mg/kg/day i.p.) for 12 weeks resulted in a lowering of blood pressure (250 +/- 9 versus 284 +/- 8 mm Hg), renal expression of transforming growth factor-beta mRNA (1.5 +/- 0.2 versus 5.4 +/- 1.4) and the matrix components: plasminogen activator inhibitor-1 (5.2 +/- 1.4 versus 31.4 +/- 10.7), fibronectin (2.2 +/- 0.6 versus 8.2 +/- 2.2), collagen I-alpha 1 (5.6 +/- 2.0 versus 23.8 +/- 7.3), and collagen III (2.7 +/- 0.9 versus 7.6 +/- 2.1). Data were corrected for rpL32 mRNA expression and expressed relative to Wistar Kyoto (WKY) rats [=1.0]. Expression of fibronectin protein was also lowered by eprosartan (0.8 +/- 0.1 versus 1.9 +/- 0.5), relative to WKY rats. Eprosartan provided significant renoprotection to SHR-SP rats as measured by decreased proteinuria (22 +/- 2 versus 127 +/- 13 mg/day) and histological evidence of active renal damage (5 +/- 2 versus 195 +/- 6) and renal fibrosis (5.9 +/- 0.7 versus 16.4 +/- 1.9) in vehicle- versus eprosartan-treated rats, respectively. Our results demonstrated that AT(1) receptor blockade with eprosartan can reduce blood pressure and preserve renal structure and function in this model of severe, chronic hypertension. These effects were accompanied by a decreased renal expression of transforming growth factor-beta1, plasminogen activator inhibitor 1, and several other extracellular matrix proteins compared with vehicle-treated SHR-SP. PMID- 11907154 TI - Prevention of in vivo excitotoxicity by a family of trialkylglycines, a novel class of neuroprotectants. AB - Excitotoxicity has been implicated in the etiology of ischemic stroke and chronic neurodegenerative disorders. Hence, the development of novel neuroprotectant molecules that ameliorate excitotoxic brain damage is vigorously pursued. We used a neuroprotection-based cellular assay to screen a synthetic combinatorial library of N-alkylglycine trimers. Two compounds (6-1-2 and 6-1-10) that efficiently prevented excitotoxic neurodegeneration in vitro and in vivo were identified. Both molecules protected primary cultures of cerebellar neurons against glutamate-induced neuronal death with an efficiency equivalent to N methyl-D-aspartate (NMDA) receptor antagonists. These trialkylglycines did not block appreciably the NMDA receptor channel, or attenuated glutamate-induced increase of Ca(2+), or affect the glutamate-nitric oxide-cGMP pathway. Intraperitoneal injection of both peptoids in mice attenuated > or = 80% ammonia induced, NMDA receptor-mediated animal death. Furthermore, these two molecules reduced by > or = 50% the neurodegeneration in striatum in a rat model of cerebral ischemia. Neuroprotection against ischemia was associated with decreased activation of caspase-3, reflecting prevention of apoptotic neuronal death. Collectively, the results reported indicate that these trialkylglycines are new neuroprotectant leads with important in vivo activity against excitotoxicity, and that they act on a novel, yet-unrecognized cellular target. These lead compounds may become tolerated drugs for the treatment of acute and chronic neurodegenerative diseases with fewer side effects than NMDA receptor antagonists. PMID- 11907155 TI - Elucidation of vasoactive intestinal peptide pharmacophore for VPAC(1) receptors in human, rat, and guinea pig. AB - Vasoactive intestinal peptide (VIP) is a neurotransmitter involved in a number of pathological and physiological processes. VIP is rapidly degraded and simplified stable analogs are needed. VIP's action was extensively studied in rat and guinea pig. However, it is largely unknown whether its pharmacophore in these species resembles human. To address this issue we investigated the VIP pharmacophore for VPAC(1) (the predominant receptor subtype in cancers and widely distributed in normal tissues) by using alanine and D-amino acid scanning. Interaction with rat, guinea pig, and human VPAC(1) was assessed using transfected Chinese hamster ovary (CHO) and PANC1 cells and cells possessing native VPAC(1). Important species differences existed in the VIP pharmacophore. The human VPAC(1) expressed in CHO cells, which were used almost exclusively in previous studies, differed markedly from the native VPAC(1) in T47D cells. The most important amino acids for determining affinity are His(1), Asp(3), Phe(6), Arg(12), Arg(14), and Leu(23). Ser(2), Asp(8), Asn(9), Thr(11), Val(19), Asn(24), Ser(25), Leu(27), and Asn(28) are not essential for high-affinity interaction/activation. [Ala(2,8,9,11,19,24,25,27,28)]VIP, which contained 11 alanines, was synthesized and it was equipotent to VIP at VPAC(1) receptors in all species and was metabolically stable. Our results show in any design of simplified VIP analogs for VPAC(1) it will be important to consider species differences and it is essential to use transfected systems that reflect the native receptor's pharmacophore. Last, with our results a simplified, metabolically stable VIP analog was identified that should be useful as a prototype for design of selective agonists/antagonists that could be useful therapeutically. PMID- 11907156 TI - Beta(1)-selective agonist (-)-1-(3,4-dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2 propanol [(-)-RO363] differentially interacts with key amino acids responsible for beta(1)-selective binding in resting and active states. AB - (-)-1-(3,4-Dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanol [(-)-RO363] is a highly selective beta(1)-adrenergic receptor (beta(1)AR) agonist. To study the binding site of beta(1)-selective agonist, chimeric beta(1)/beta(2)ARs and Ala substituted beta(1)ARs were constructed. Several key residues of beta(1)AR [Leu(110) and Thr(117) in transmembrane domain (TMD) 2], and Phe(359) in TMD 7] were found to be responsible for beta(1)-selective binding of (-)-RO363, as determined by competitive binding. Based on these results, we built a three dimensional model of the binding domain for (-)-RO363. The model indicated that TMD 2 and TMD 7 of beta(1)AR form a binding pocket; the methoxyphenyl group of N substituent of (-)-RO363 seems to locate within the cavity surrounded by Leu(110), Thr(117), and Phe(359). The amino acids Leu(110) and Phe(359) interact with the phenyl ring of (-)-RO363, whereas Thr(117) forms hydrogen bond with the methoxy group of (-)-RO363. To examine the interaction of these residues with beta(1)AR in an active state, each of the amino acids was changed to Ala in a constitutively active (CA)-beta(1)AR mutant. The degree of decrease in the affinity of CA-beta(1)AR for (-)-RO363 was essentially the same as that of wild type beta(1)AR when mutated at Leu(110) and Thr(117). However, the affinity was decreased in Ala-substituted mutant of Phe(359) compared with that of wild-type beta(1)AR. These results indicated that Leu(110) and Thr(117) are necessary for the initial binding of (-)-RO363 with beta(1)-selectivity, and interaction of Phe(359) with the N-substituent of (-)-RO363 in an active state is stronger than in the resting state. PMID- 11907157 TI - New cytokine delivery system using gelatin microspheres containing interleukin-10 for experimental inflammatory bowel disease. AB - Interleukin (IL)-10 is an anti-inflammatory cytokine that suppresses the T helper 1 immune response and down-regulates macrophages and monocytes. The therapeutic effect of systemic administration of IL-10 for patients with inflammatory bowel disease, however, has not been satisfactory. We examined whether rectal administration of gelatin microspheres (GM) containing IL-10 (GM-IL-10) prevents colitis in IL-10-deficient (IL-10(-/-)) mice. GM-IL-10 and IL-10 alone were administered rectally. The colon was examined macroscopically and microscopically. IL-12 mRNA expression and CD40 expression in Mac-1-positive cells were also examined. Macroscopic and microscopic examination revealed marked improvement of colitis in IL-10(-/-) mice treated with GM-IL-10. mRNA expression of IL-12 in Mac-1-positive cells in GM-IL-10-treated mice was significantly decreased compared with that in the mice treated with IL-10 alone. Additionally, CD40 expression in Mac-1-positive cells in GM-IL-10-treated mice was decreased more prominently than in mice treated with IL-10 alone. The therapeutic effects of GM-IL-10 were associated with decreased expression of IL-12 mRNA and down regulation of CD40 expression in Mac-1-positive cells. GM-IL-10 might be useful for treatment of patients with inflammatory bowel disease. PMID- 11907158 TI - Ethanol stimulates cAMP-responsive element (CRE)-mediated transcription via CRE binding protein and cAMP-dependent protein kinase. AB - Alcoholism is characterized by tolerance, dependence, and unrestrained craving for alcohol. Adaptive responses, including changes in gene expression in neurons, are thought to account for some of these complex behavioral abnormalities. We have shown in the NG108-15 neuroblastoma x glioma hybrid cell line that ethanol increases cellular cAMP levels via activation of adenosine A(2) receptors, leading to phosphorylation of the cAMP response element-binding protein (CREB). However, phosphorylation of CREB is not sufficient to activate cAMP response element (CRE)-mediated gene expression. Here we investigate whether ethanol increases CRE-mediated gene expression via endogenous CREB using a CRE-regulated luciferase reporter construct, transfected into NG108-15 cells. We find increased luciferase activity as a function of time of exposure to ethanol. Coexpression of a dominant-negative CREB construct blocked ethanol-stimulated CRE-luciferase expression, further suggesting that CREB is required for this response. We also determined whether ethanol-induced increases in gene expression are mediated by ethanol-induced increases in extracellular adenosine. We found that CRE-mediated gene expression induced by ethanol occurs in two phases: an early phase (4 h), in which adenosine receptor blockade prevents ethanol-induced gene expression, and a later phase (14 h), which is not blocked by an adenosine receptor antagonist. In both phases, inhibition of cAMP-dependent protein kinase A (PKA) activity prevented ethanol-induced CRE-mediated luciferase expression. Our data suggest that ethanol induces cAMP-dependent gene expression regulated by CREB and PKA and that this signaling pathway may mediate some of the addictive behaviors underlying alcoholism. PMID- 11907159 TI - Differential effects of bucindolol and carvedilol on noradenaline-induced hypertrophic response in ventricular cardiomyocytes of adult rats. AB - In adult rat ventricular cardiomyocytes, noradrenaline exerts dual effects on protein synthesis: increases via alpha(1)-adrenoceptors and decreases via beta(1) adrenoceptors. Carvedilol and bucindolol are beta-blockers with additional alpha(1)-adrenoceptor blocking activities. We studied the effects of carvedilol and bucindolol on noradrenaline-induced protein synthesis (assessed by [(3)H]phenylalanine incorporation) in adult rat ventricular cardiomyocytes. Radioligand binding studies with [(125)I]iodocyanopindolol and [(3)H]prazosin revealed that carvedilol had a much higher affinity to alpha(1)-adrenoceptors than bucindolol (beta(1)-/alpha(1)-adrenoceptor ratio for carvedilol, 1:2.7; for bucindolol, 1:43). Noradrenaline-evoked increases in protein synthesis were enhanced by propranolol (1 microM) and beta(1)-adrenoceptor-selective antagonists bisoprolol (1 microM) and CGP 20712A [1-[2-((3-carbamoyl-4-hydroxy)phenoxy)-ethyl amino]-3-[4-(1-methyl-4-trifluoromethyl-2-imidazolyl)phenoxy]-2-propranol methanesulfonate] (300 nM). Carvedilol (100 pM-10 microM) inhibited 1 microM noradrenaline-induced increase in protein synthesis with monophasic concentration inhibition curves independent of whether CGP 20712A was present or not; K(i) values for carvedilol were 5 to 6 nM. In contrast, bucindolol (100 pM-10 microM) inhibited l microM noradrenaline-induced increase in protein synthesis with a bell-shaped concentration-inhibition curve; it increased noradrenaline-induced protein synthesis at 10 nM, although at concentrations >100 nM it was inhibited. In the presence of 300 nM CGP 20712A or 1 microM propranolol, however, bucindolol inhibited 1 microM noradrenaline-induced increase in protein synthesis with monophasic concentration-inhibition curves; K(i) values were 40 to 75 nM. On the other hand, both carvedilol and bucindolol inhibited 1 microM phenylephrine induced protein synthesis with monophasic concentration-inhibition curves; K(i) values were 4 (carvedilol) and 45 nM (bucindolol). These results indicate that, at low (beta-adrenoceptor blocking) concentrations, bucindolol can enhance noradrenaline-induced protein synthesis whereas it is inhibited by carvedilol. PMID- 11907160 TI - Novel glutathione-dependent thiopurine prodrugs: evidence for enhanced cytotoxicity in tumor cells and for decreased bone marrow toxicity in mice. AB - Elevated glutathione (GSH) levels have been detected in many tumors compared with the healthy, surrounding tissue. Often, this GSH up-regulation is associated with drug resistance. The prodrugs 6-(2-acetylvinylthio)guanine (AVTG) and 6-(2 acetylvinylthio)purine (AVTP) contain a novel butenone moiety that allows the prodrugs to react selectively with sulfhydryl nucleophiles to release the chemotherapeutic drug 6-thioguanine (6-TG) or 6-mercaptopurine (6-MP), respectively. The cellular uptake and metabolism of trans-AVTG in two human renal carcinoma cell lines that were used as models were rapid and associated with depletion of intracellular GSH. Formation of 6-TG from trans-AVTG correlated positively with intracellular GSH concentrations, and was significantly reduced by diethyl maleate pretreatment. Intracellular concentrations of 6-TG after incubations with trans-AVTG were significantly higher than the 6-TG concentrations obtained after incubations with equimolar concentrations of 6-TG; thus, the prodrug delivered more 6-TG to the cell than did 6-TG itself. Cytotoxicity studies demonstrated that AVTG and AVTP had similar IC(50) values that were comparable with those of 6-TG, but were significantly lower than those of 6-MP. Furthermore, after in vivo treatment of mice with the prodrugs, no reduction was observed in circulating white blood cell counts, whereas white blood cell counts of mice treated with equimolar or 60% lower doses of 6-TG were reduced by 50 to 60%. Collectively, the results show that AVTG and AVTP are novel potential chemotherapeutic agents that may target tumors with up-regulated levels of GSH, and may exhibit less systemic toxicity than the parent thiopurines. PMID- 11907161 TI - Priming of alveolar macrophage respiratory burst by H(2)O(2) is prevented by phosphatidylcholine-specific phospholipase C inhibitor Tricyclodecan-9-yl xanthate (D609). AB - The respiratory burst in alveolar macrophages is enhanced in vitro by pre exposure to nontoxic concentrations of hydroperoxides before stimulation by an agonist, which may represent a feed-forward regulatory mechanism. Tricyclodecan-9 yl-xanthate (D609), an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), suppresses this priming of the respiratory burst by pre-exposure to H(2)O(2) in NR8383 alveolar macrophages (up to 100 microM D609, 400 nmol of H(2)O(2) added to 5 x 10(6) cells 15 min before stimulation with ADP). D609 has potential as an antioxidant due to its dithiocarbonate functional group that allows it to slowly react with H(2)O(2) and rapidly reduce cytochrome c, which interferes with a common assay for the respiratory burst. Nonetheless, the antioxidant properties of D609 do not account for its inhibition of priming of the respiratory burst by H(2)O(2). Reduction of nitro blue tetrazolium is the basis for an assay for superoxide production with which D609 does not interfere. With this assay, it was found that D609 does not inhibit the respiratory burst per se, but prevents its enhancement by pre-exposure to H(2)O(2). Consistent with a role of diacylglycerol generation by phospholipase C, this enhancement was mimicked by pre-exposure to phorbol ester. In contrast with priming, receptor mediated stimulation of the respiratory burst depends on the better characterized phosphatidylinositol-specific phospholipase C. Priming of the respiratory burst by H(2)O(2) joins the list of inflammatory responses that are inhibited by D609. Nevertheless, the results herein indicate that caution should be exercised in the interpretation of the effects of D609 to consider both antioxidant effects and inhibition of PC-PLC. PMID- 11907162 TI - Pharmacological profile of a new, potent, and long-acting gonadotropin-releasing hormone antagonist: degarelix. AB - We describe the pharmacological profile in rats and monkeys of degarelix (FE200486), a member of a new class of long-acting gonadotropin-releasing hormone (GnRH) antagonists. At single subcutaneous injections of 0.3 to 10 microg/kg in rats, degarelix produced a dose-dependent suppression of the pituitary-gonadal axis as revealed by the decrease in plasma luteinizing hormone (LH) and testosterone levels. Duration of LH suppression increased with the dose: in the rat, significant suppression of LH lasted 1, 2, and 7 days after a single subcutaneous injection of degarelix at 12.5, 50, or 200 microg/kg, respectively. Degarelix fully suppressed plasma LH and testosterone levels in the castrated and intact rats as well as in the ovariectomized rhesus monkey for more than 40 days after a single 2-mg/kg subcutaneous injection. In comparative experiments, degarelix showed a longer duration of action than the recently developed GnRH antagonists abarelix, ganirelix, cetrorelix, and azaline B. The in vivo mechanism of action of degarelix was consistent with competitive antagonism, and the prolonged action of degarelix was paralleled by continued presence of radioimmunoassayable degarelix in the general circulation. In contrast to cetrorelix and similarly to ganirelix and abarelix, degarelix had only weak histamine-releasing properties in vitro. These results demonstrate that the unique and favorable pharmacological properties of degarelix make it an ideal candidate for the management of sex steroid-dependent pathologies requiring long term inhibition of the gonadotropic axis. PMID- 11907163 TI - Inhibitors of ATP-binding cassette transporters suppress interleukin-12 p40 production and major histocompatibility complex II up-regulation in macrophages. AB - ATP-binding cassette (ABC) transporters are a large family of proteins whose role is to translocate various substances across biological membranes. They include the Tangier disease protein ABC1, sulfonylurea receptors (SUR), multidrug resistance protein (MDR), and cystic fibrosis transmembrane regulator (CFTR). In the current study, we investigated the involvement of ABC transporters in the regulation of lipopolysaccharide (LPS) and/or interferon (IFN)-gamma-induced interleukin (IL)-12 p40 and tumor necrosis factor (TNF)-alpha production, nitric oxide formation, as well as major histocompatibility complex II up-regulation in macrophages. The general ABC transporter inhibitor glibenclamide suppressed both IL-12 p40 and nitric oxide production. However, glibenclamide failed to affect the production of TNF-alpha. The selective ABC1 inhibitors 4,4' diisothiocyanostilbene-2,2'-disulfonic acid and sulfobromophthalein mimicked the suppressive effect of glibenclamide on IL-12 p40 production. On the other hand, both the MDR inhibitor verapamil and CFTR blocker 2,2'-iminodibenzoic acid failed to suppress the production of IL-12 p40. Furthermore, selective inhibitors and activators of SURs were without effect. In agreement with the pharmacological data, macrophages expressed mRNA for ABC1, but not SURs or CFTR. Intracellular levels of IL-12 p40 were decreased by glibenclamide, suggesting that glibenclamide does not affect IL-12 p40 secretion. The effect of glibenclamide did not involve an interference with the activation of the p38 and p42/44 mitogen activated protein kinases or c-Jun kinase. Glibenclamide also suppressed IFN gamma-induced up-regulation of major histocompatibility complex II. Taken together, our results indicate that ABC proteins regulate LPS and/or IFN-gamma induced macrophage activation. PMID- 11907164 TI - Adenovirus-mediated overexpression of catalase in the cytosolic or mitochondrial compartment protects against toxicity caused by glutathione depletion in HepG2 cells expressing CYP2E1. AB - Induction of cytochrome P450 CYP2E1 by ethanol appears to be one of the mechanisms by which ethanol creates a state of oxidative stress. Glutathione (GSH) is a key cellular antioxidant that detoxifies reactive oxygen species. Depletion of GSH, especially mitochondrial GSH, is believed to play a role in the ethanol-induced liver injury. Previous results reported that depletion of GSH by buthionine-(S,R)-sulfoximine (BSO) treatment caused apoptosis and necrosis in HepG2 cells, which overexpress CYP2E1. In the current work, adenoviral infection with vectors that resulted in expression of catalase either in the cytosol or mitochondrial compartments was able to abolish the loss of mitochondrial membrane potential or damage to mitochondria observed in HepG2 cells overexpressing CYP2E1 that were treated with BSO. Loss of cell viability, either necrotic or apoptotic, was also prevented by the catalase overexpression after infection with the adenoviral vectors. The protective effects of catalase were associated with the suppression of the increase in the production of reactive oxygen species and of mitochondrial lipid peroxidation observed after GSH depletion. These results reveal a prominent role for H(2)O(2) as a mediator in the cytotoxicity observed after depletion of GSH in HepG2 cells overexpressing CYP2E1. Damage to mitochondria may be a critical step for cellular toxicity by CYP2E1-derived reactive oxygen species. PMID- 11907165 TI - Role of protein kinase C in control of ethanol-modulated beta-endorphin release from hypothalamic neurons in primary cultures. AB - We have previously shown that short-term exposure to ethanol stimulates immunoreactive beta-endorphin (IR-beta-EP) release from hypothalamic neurons and that chronic ethanol exposure decreases the IR-beta-EP release from these neurons. The role of protein kinase C (PKC) in the ethanol-regulated beta-EP release from hypothalamic neurons has not been established. In this study, by using the primary cultures of hypothalamic neurons, we tested the effects of PKC stimulator phorbol ester 4 beta-phorbol 12-myristate-13-acetate (PMA) and PKC inhibitor chelerythrine chloride on ethanol-induced IR-beta-EP release. Additionally, the effects of ethanol with or without PMA on expression and translocation of various PKC isoenzymes from cytosolic to membrane fraction were determined. PMA treatment increased IR-beta-EP release in a time- and dose dependent manner. Acute ethanol treatment (3 h) increased, while chronic ethanol treatment (24 h) reduced, the magnitude of PMA-induced IR-beta-EP release. The stimulatory effect of acute ethanol on IR-beta-EP release was reduced by chelerythrine chloride. Determination of the effects of ethanol with or without PMA on seven different PKC isoenzymes (PKC-alpha, -beta I, -beta II, -gamma, delta, -epsilon, and -zeta) revealed that the expression and translocation of only two PKC isoenzymes, PKC-delta and PKC-epsilon, were stimulated by acute treatment with ethanol. Acute ethanol also increased PMA-stimulated expression of these two isoenzymes. Chronic ethanol treatment reduced both basal and PMA induced increase of PKC-delta and PKC-epsilon expression and translocation. These data provide evidence for the first time that ethanol-regulated IR-beta-EP secretion is controlled by the PKC system, possibly involving PKC-delta and PKC epsilon isoenzymes. PMID- 11907167 TI - 3,4-Methylenedioxymethamphetamine produces glycogenolysis and increases the extracellular concentration of glucose in the rat brain. AB - Oxidative and/or bioenergetic stress is thought to contribute to the mechanism of neurotoxicity of amphetamine derivatives, e.g., 3,4-methylenedioxymethamphetamine (MDMA). In the present study, the effect of MDMA on brain energy regulation was investigated by examining the effect of MDMA on brain glycogen and glucose. A single injection of MDMA (10-40 mg/kg, s.c.) produced a dose-dependent decrease (40%) in brain glycogen, which persisted for at least 1 h. MDMA (10 and 40 mg/kg, s.c.) also produced a significant and sustained increase in the extracellular concentration of glucose in the striatum. Subjecting rats to a cool ambient temperature of 17 degrees C significantly attenuated MDMA-induced hyperthermia and glycogenolysis. MDMA-induced glycogenolysis also was prevented by treatment of rats with the 5-hydroxytryptamine(2) (5-HT(2)) antagonists 6-methyl-1-(1 methylethyl)-ergoline-8 beta-carboxylic acid 2-hydroxy-1 methylprophyl ester maleate (LY-53,857; 3 mg/kg i.p.), desipramine (10 mg/kg i.p.), and iprindole (10 mg/kg i.p.). LY-53,857 also attenuated the MDMA-induced increase in the extracellular concentration of glucose as well as MDMA-induced hyperthermia. Amphetamine analogs (e.g., methamphetamine and parachloroamphetamine) that produce hyperthermia also produced glycogenolysis, whereas fenfluramine, which does not produce hyperthermia, did not alter brain glycogen content. These results support the conclusion that MDMA induces glycogenolysis and that the process involves 5-HT(2) receptor activation. These results are supportive of the view that MDMA promotes energy dysregulation and that hyperthermia may play an important role in MDMA-induced alterations in cellular energetics. PMID- 11907166 TI - Altered cardiovascular responses in mice lacking the M(1) muscarinic acetylcholine receptor. AB - Although the M(2) muscarinic acetylcholine receptor (mAChR) is the predominant functional mAChR subtype in the heart, some responses of the cardiovascular system to acetylcholine (ACh) may be mediated by other mAChR subtypes. The potential effect of M(1) mAChR on heart function was investigated using M(1) knockout (M(1)-KO) mice. In vivo cardiodynamic analysis showed that basal values of heart rate (HR), developed left ventricular pressure (DLVP), left ventricular dP/dt(max) (LV dP/dt(max)), and mean blood pressure (MBP) were similar between wild-type (WT) and M(1)-KO mice. Injection of the putative M(1)-selective agonist 4-(m-chlorophenyl-carbamoyloxy)-2-butynyltrimethylammonium (McN-A-343) produced an increase in LV dP/dt(max), DLVP, HR, and MBP in WT mice but did not affect hemodynamic function in the M(1)-KO mice. The stimulatory effect of McN-A-343 in WT mice was blocked by pretreatment with propranolol, indicating that stimulation of the M(1) mAChRs on sympathetic postganglionic neurons evoked release of catecholamines. Intravenous injection of ACh in both WT and M(1)-KO mice caused atrioventricular conduction block, without a significant change in the frequency of atrial depolarization, or atrial fibrillation. Immunoprecipitation and reverse transcriptase-polymerase chain reaction failed to detect the expression of M(1) mAChR in cardiac tissue from WT mice. The carbachol-induced increase of phospholipase C activity in cardiac tissues was not different between WT and M(1) KO mice. These results demonstrate that 1) activation of M(1) mAChR subtype on sympathetic postganglionic cells results in catecholamine-mediated cardiac stimulation, 2) M(1) mAChR is not expressed in mouse heart, and 3) administration of ACh to mice induces arrhythmia. PMID- 11907168 TI - Gender-specific and developmental influences on the expression of rat organic anion transporters. AB - Rat organic anion transporter 1 (Oat1), Oat2, and Oat3, members of the organic anion transporter family, transport some organic anions across cellular membranes. Previously, highest Oat1 and Oat3 mRNA expression was reported in kidney and Oat2 in liver. However, gender and developmental differences in Oat expression remain unknown. This study describes gender- and age-specific patterns of rat organic anion transporter expression in various tissues. Oat mRNA expression was evaluated in adult male and female Sprague-Dawley rat tissues, and developmental expression was also determined in kidneys of Sprague-Dawley rats ranging in age from days 0 through 45. Expression was quantified using branched DNA signal amplification. Oat1 mRNA expression was primarily observed in kidney. Surprisingly, Oat2 mRNA expression was also highest in kidney rather than in liver. Moreover, considerably higher Oat2 levels were seen in female kidney as compared with male. Finally, Oat3 mRNA expression was highest in kidney of both genders, whereas a male-predominant pattern was observed in liver. At birth, all kidney Oat mRNA levels were low. Renal Oat1 expression gradually increased throughout development, approaching adult levels at 30 days of age, where at days 40 and 45 Oat1 levels were greater in males than females. Oat2 expression in kidney was minimal through day 30 but increased dramatically at day 35 in females only. Lastly, Oat3 mRNA expression in kidney matured earliest, rapidly increasing from birth through day 10. These data indicate that Oat mRNA expression is primarily localized to the kidney, and observed expression patterns may explain some previously recognized age- and gender-dependent toxicities associated with chemical exposure. PMID- 11907169 TI - Cardiovascular responses elicited by the "binge" administration of methamphetamine. AB - Methamphetamine (METH) abuse is often characterized by a repeated pattern of frequent drug administrations (binge) followed by a period of abstinence. The effect of this pattern of METH use on cardiovascular function has not been characterized. Radiotelemetry was used to record the cardiovascular responses elicited during three successive METH binges (3 mg/kg, b.i.d. for 4 days) in conscious rats. Each binge was followed by a 10-day METH-free period. The effects of METH administration on vascular reactivity, Bezold-Jarisch reflex function, and cardiac morphology were also evaluated. The pressor responses elicited by the first three doses of METH in the second and third binges were significantly larger than those elicited by the corresponding doses in the first binge. The heart rate (HR) responses elicited by METH were similar within and among the three binges. Ten days after the last binge, the depressor responses elicited by the i.v. injection of sodium nitroprusside, isoproterenol, and acetylcholine were significantly smaller than those elicited before each binge. The arterial pressure and HR responses elicited by phenylephrine were unchanged. Bezold Jarisch reflex function evoked by i.v. serotonin (10 microg/kg) was significantly altered. The hearts from treated rats showed focal inflammatory infiltrates with abundant monocytes and occasional necrotic foci. These results indicate that this binge pattern of METH administration can significantly alter cardiovascular function and cardiovascular reflex function and produce serious cardiac pathology. PMID- 11907170 TI - Mechanism-based inactivation of cytochrome P450 3A4 by 17 alpha-ethynylestradiol: evidence for heme destruction and covalent binding to protein. AB - 17 alpha-Ethynylestradiol (EE), a major constituent of many oral contraceptives, inactivated the testosterone 6 beta-hydroxylation activity of purified P450 3A4 reconstituted with phospholipid and NADPH-cytochrome P450 reductase in a mechanism-based manner. The inactivation of P450 3A4 followed pseudo first order kinetics and was dependent on NADPH. The values for the K(I) and k(inact) were 18 microM and 0.04 min(-1), respectively, and the t(1/2) was 16 min. Incubation of 50 microM EE with P450 3A4 at 37 degrees C for 30 min resulted in a 67% loss of testosterone 6 beta-hydroxylation activity accompanied by a 35% loss of the spectral absorbance of the native protein at 415 nm and a 70% loss of the spectrally detectable P450-CO complex. The inactivation of P450 3A4 by EE was irreversible. Testosterone, an alternate substrate, was able to protect P450 3A4 from EE-dependent inactivation. The partition ratio was approximately 50. The stoichiometry of binding was approximately 1.3 nmol of an EE metabolite bound per nmol of P450 3A4 inactivated. SDS-polyacrylamide gel electrophoresis analysis demonstrated that [(3)H]EE was irreversibly bound to the P450 3A4 apoprotein. After extensive dialysis of the [(3)H]EE inactivated samples, high-pressure liquid chromatography (HPLC) analysis demonstrated that the inactivation resulting from EE metabolism led to the destruction of approximately half the heme with the concomitant generation of modified heme and EE-labeled heme fragments and produced covalently radiolabeled P450 3A4 apoprotein. Electrospray mass spectrometry demonstrated that the fraction corresponding to the major radiolabeled product of EE metabolism has a mass (M - H)(-) of 479 Da. HPLC and gas chromatography-mass spectometry analyses revealed that EE metabolism by P450 3A4 generated one major metabolite, 2-hydroxyethynylestradiol, and at least three additional metabolites. In conclusion, our results demonstrate that EE is an effective mechanism-based inactivator of P450 3A4 and that the mechanism of inactivation involves not only heme destruction, but also the irreversible modification of the apoprotein at the active site. PMID- 11907171 TI - Tranexamic acid, a widely used antifibrinolytic agent, causes convulsions by a gamma-aminobutyric acid(A) receptor antagonistic effect. AB - Application of 4-(aminomethyl)cyclohexanecarboxylic acid (tranexamic acid; TAMCA) to the central nervous system (CNS) has been shown to result in hyperexcitability and convulsions. However, the mechanisms underlying this action are unknown. In the present study, we demonstrate that TAMCA binds to the gamma-aminobutyric acid (GABA) binding site of GABA(A) receptors in membranes from rat cerebral cortex and does not interfere with N-methyl-D-aspartate receptors. Patch-clamp studies using human embryonic kidney cells transiently transfected with recombinant GABA(A) receptors composed of alpha 1 beta 2 gamma 2 subunits showed that TAMCA did not activate these receptors but dose dependently blocked GABA-induced chloride ion flux with an IC(50) of 7.1 +/- 3.1 mM. Application of TAMCA to the lumbar spinal cord of rats resulted in dose-dependent hyperexcitability, which was completely blocked by coapplication of the GABA(A) receptor agonist muscimol. These results indicate that TAMCA may induce hyperexcitability by blocking GABA driven inhibition of the CNS. PMID- 11907172 TI - Identification of a novel route of extraction of sirolimus in human small intestine: roles of metabolism and secretion. AB - Using Caco-2 cell monolayers expressing CYP3A4, we investigated the interplay between metabolism and transport on the first-pass intestinal extraction of the immunosuppressant sirolimus, a CYP3A4/P-glycoprotein (P-gp) substrate. Modified Caco-2 cells metabolized [(14)C]sirolimus to the predicted amounts of CYP3A4 mediated products based on CYP3A4 content, which was approximately 20% of that measured in human small intestinal mucosal homogenate. [(14)C]Sirolimus also degraded to the known ring-opened product, seco-rapamycin. Unexpectedly, a ring opened dihydro metabolite (M2) was the major product detected in cells at all sirolimus concentrations examined (2-100 microM). Greater M2 formation after apical versus basolateral dosing (1.6-fold) was explained by higher intracellular content of sirolimus after apical dosing. M2 was not detected in incubations with human liver and intestinal microsomes but was readily detected with corresponding homogenates. M2 formation was NADPH-dependent but unaffected by the CYP3A4 inhibitors ketoconazole and troleandomycin. Although M2 was formed from purified seco-rapamycin (20 microM) in the homogenates, it was not detected in cells when seco-rapamycin was added to the apical compartment, because seco-rapamycin was essentially impermeable to the apical membrane. Sirolimus, seco-rapamycin (basolaterally dosed), and M2 were all actively secreted across the apical membrane, and secretion of each was inhibited by the P-gp inhibitor LY335979 [(2R)-anti-5-[3-[4-(10,11-difluoromethanodibenzo-suber-5-yl)piperazin-1-yl]-2 hydroxypropoxy]quinoline trihydrochloride]. Along with CYP3A4-mediated metabolism and P-gp-mediated secretion, we conclude that the following novel pathway, which occurs at least in the intestine, may contribute significantly to the first-pass extraction of sirolimus in humans: intracellular degradation of sirolimus to seco rapamycin, metabolism of seco-rapamycin to M2 by an unidentified non-microsomal enzyme, and P-gp-mediated secretion of M2 and seco-rapamycin. PMID- 11907173 TI - Effects of acute and subchronic administration of dexefaroxan, an alpha(2) adrenoceptor antagonist, on memory performance in young adult and aged rodents. AB - The present study examined the influence of dexefaroxan, a potent and selective alpha(2)-adrenoceptor antagonist, on cognitive performance in rodents. In young adult rats, dexefaroxan reversed the deficits induced by UK 14304 [5-bromo-N-(4,5 dihydro-1-H-imidazol-2-yl)-6-quinoxalinamine], scopolamine, and diazepam in a passive avoidance task. In this test, dexefaroxan also attenuated the spontaneous forgetting induced by a 15-week training-testing interval. Moreover, dexefaroxan, given immediately after training, increased the memory performance of rats trained with a weak electric footshock in the passive avoidance test, facilitated spatial memory processes in the Morris water maze task in rats, and increased the performance of mice in an object recognition test. Thus, dexefaroxan appears to have a promnesic effect in these tests by facilitating the processes of memory retention, rather than acquisition or other noncognitive influences. The facilitatory effects of dexefaroxan in young adult rats persisted even after a 21 to 25-day constant subcutaneous infusion by using osmotic minipumps, indicating that tolerance to the promnesic effect of the drug did not occur during this prolonged treatment interval. Furthermore, in the passive avoidance and Morris water maze tests, dexefaroxan ameliorated the age-related memory deficits of 24 month-old rats to a level that was comparable to that of young adult animals, and reversed the memory deficits induced by excitotoxin lesions of the nucleus basalis magnocellularis region. Together, these findings support a potential utility of dexefaroxan in the treatment of cognitive deficits occurring in Alzheimer's disease. PMID- 11907174 TI - Differential effects of haloperidol and clozapine on [(3)H]cAMP binding, protein kinase A (PKA) activity, and mRNA and protein expression of selective regulatory and catalytic subunit isoforms of PKA in rat brain. AB - The present study was undertaken to examine whether the mechanism of action of typical and atypical antipsychotics is related in their ability to regulate key phosphorylating enzyme of adenylyl cyclase-cAMP pathway, i.e., protein kinase A (PKA). For this purpose, regulatory (R) and catalytic (Cat) activities of PKA and expression of various isoforms of regulatory and catalytic subunits were examined in rat brain after single or chronic (21-day) treatment with haloperidol (HAL, 1 mg/kg) or clozapine (CLOZ, 20 mg/kg). It was observed that chronic but not acute treatment of CLOZ significantly decreased [(3)H]cAMP binding to the regulatory subunit of PKA as well as catalytic activity of PKA in particulate and cytosol fractions of the rat cortex, hippocampus, and striatum. In these fractions, CLOZ significantly decreased protein levels of selective RII alpha-, RII beta-, and Cat beta-subunit isoforms of PKA. These decreases were accompanied by decreases in their respective mRNA expression. In contrast, chronic but not acute treatment of HAL significantly increased [(3)H]cAMP binding and the catalytic activity of PKA in particulate and cytosol fractions of only the striatum brain area. In addition, chronic treatment of HAL significantly increased mRNA and protein levels of RII alpha- and RII beta-subunit isoforms in the striatum. None of the antipsychotics caused any change in the expression of the Cat alpha-, RI alpha-, or RI beta-subunit isoform. These results, thus, suggest that HAL and CLOZ differentially regulate PKA catalytic and regulatory activities and the expression of selective catalytic and regulatory subunit isoforms of PKA, which may be associated with their mechanisms of action. PMID- 11907175 TI - Neuroprotective effect of (2S,3S,4R)-N"-cyano-N-(6-amino-3, 4-dihydro-3-hydroxy-2 methyl-2-dimethoxymethyl-2H-benzopyran-4-yl)-N'-benzylguanidine (KR-31378), a benzopyran analog, against focal ischemic brain damage in rats. AB - This study shows the preventive effect of KR-31378 [(2S,3S,4R)-N"-cyano-N-(6 amino-3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2H-benzopyran-4-yl)-N' benzylguanidine] against cerebral infarct via antioxidant and antiapoptotic actions evoked by subjecting rats to 2 h of occlusion of the left middle cerebral artery followed by 24 h of reperfusion. The brain infarct zone in the cortex and striatum of the left hemisphere was consistently identified in the cortex and striatum of the left hemisphere. The infarct area was significantly reduced after three intraperitoneal administrations of 10, 30, or 50 mg/kg KR-31378 at 5 min, 4 h, and 8 h after the completion of 2 h of ischemia. Treatment with KR-31378 (30 or 50 mg/kg) significantly reduced the increase in the number of terminal deoxynucleotidyl transferase dUTP nick-end labeling positive cells as well as strongly suppressed the laddered feature of DNA fragmentation in the lateral cortical tissue corresponding to the penumbra. The findings of samples from penumbral zone, which showed markedly reduced Bcl-2 protein level and increased Bax protein and cytochrome c release, were wholly reversed by treatment with KR 31378. In conclusion, postischemic treatment with KR-31378 provided significant levels of cortical neuroprotection in association with inhibition of apoptotic cell death through the up-regulation of Bcl-2 expression, and the down-regulation of Bax protein and cytochrome c release. PMID- 11907176 TI - Influence of porcine Actinobacillus pleuropneumoniae infection and dexamethasone on the pharmacokinetic parameters of enrofloxacin. AB - The impact of Actinobacillus pleuropneumoniae (APP) infection in swine on the pharmacokinetic parameters of enrofloxacin were determined. Twenty-four animals were used in a 2 x 2 factorial of treatment groups (six animals per group) to determine the impact of APP-induced inflammation and the anti-inflammatory drug dexamethasone on enrofloxacin pharmacokinetic parameters. All animals received enrofloxacin as a single intravenous dose (5 mg/kg). Administration of dexamethasone was associated with an increase in clearance of enrofloxacin Clearance of enrofloxacin was not affected by APP. Volume of distribution at steady state was significantly increased in the dexamethasone-treated pigs. Volume of distribution at steady state was decreased by APP infection. Dexamethasone significantly increased the terminal elimination half-life of enrofloxacin. APP infection decreased the terminal elimination half-life of enrofloxacin in the infected pigs. Infection and dexamethasone significantly decreased the urine enrofloxacin/creatinine and ciprofloxacin/creatinine ratios. This study shows that APP infection does affect plasma pharmacokinetic parameters. Dexamethasone and APP infection may reduce renal clearance of enrofloxacin with a compensatory increase in intestinal clearance. Neither infection nor dexamethasone altered the metabolism of enrofloxacin to ciprofloxacin, the principal metabolite of enrofloxacin. PMID- 11907177 TI - Dose optimization of a doxorubicin prodrug (HMR 1826) in isolated perfused human lungs: low tumor pH promotes prodrug activation by beta-glucuronidase. AB - HMR 1826 (N-[4-beta-Glucuronyl-3-nitrobenzyl-oxycarbonyl]doxorubicin) is a nontoxic glucuronide prodrug from which active doxorubicin is released by beta glucuronidase. Preclinical studies aimed at dose optimization of HMR 1826, based on intratumoral pharmacokinetics, are important to design clinical studies. Using an isolated perfused human lung model, the uptake of doxorubicin into normal tissue and tumors after perfusion with 133 microg/ml (n = 6), 400 microg/ml (n = 10), and 1200 microg/ml (n = 6) HMR 1826 was compared. Extracellular tissue pH was measured, and enzyme kinetic studies were performed in vitro to investigate the effect of pH on the formation of doxorubicin. Extracellular pH was lower in tumors than in healthy tissue (6.46 +/- 0.35, n = 8 versus 7.30 +/- 0.33, n = 10; p < 0.001). In vitro, beta-glucuronidase activity was 10 times higher at pH 6.0 than at neutral pH. After perfusion with HMR 1826, there was a linear relationship between HMR 1826 concentrations in perfusate and normal lung tissue. After perfusion with 133, 400, and 1200 microg/ml HMR 1826, the final doxorubicin concentrations in normal and tumor tissue were 2.7 +/- 0.9, 11.1 +/- 5.4, and 21.8 +/- 8.4 microg/g (p < 0.05 for all comparisons), and 0.7 +/- 0.3, 8.6 +/- 2.0 microg/g (p < 0.01 versus 133 microg/g), and 8.7 +/- 4.9 microg/g, respectively. This agrees with the enzyme kinetic observations of saturation of beta-glucuronidase at 400 microg/ml HMR 1826 in the acidic environment of the tumor. Therefore, the escalation of the HMR 1826 dose most likely results in higher circulating concentrations than 400 microg/ml but does not increase the uptake of doxorubicin into tumors and, subsequently, antitumor efficacy. The isolated perfused human lung is an excellent model for preclinical investigations aimed at optimization of tissue pharmacokinetics of tumor-selective prodrugs. PMID- 11907178 TI - In vivo alpha(1)-adrenergic lipolytic activity in subcutaneous adipose tissue of obese subjects. AB - The role of alpha(1)-adrenoceptors in lipid mobilization and blood flow was investigated in situ using microdialysis of subcutaneous adipose tissue in severely obese subjects. The lipolysis rate was assessed by determination of interstitial glycerol concentration. The alpha(1)-adrenoceptor agonist norfenefrine caused an increase in glycerol level in adipose tissue that was similar to that observed with the physiologic alpha(1,2)-beta(1)-adrenoceptor agonist norepinephrine, whereas the alpha(1)-adrenoceptor antagonist urapidil showed no effect on basal lipolysis rate. However, the enhanced glycerol concentration due to norfenefrine and norepinephrine was suppressed in the presence of urapidil. The beta-adrenoceptor antagonist propranolol showed no effect on norfenefrine-stimulated glycerol outflow. Blood flow was assessed using the ethanol escape technique. Perfusion with norfenefrine decreased blood flow, whereas urapidil enhanced blood flow significantly. Despite the increase in blood flow, the basal interstitial glycerol concentration remained unchanged. Although norfenefrine at high concentrations could inhibit the urapidil-induced increase in blood flow, the norfenefrine-induced glycerol output was not affected. These results demonstrate that alpha(1)-adrenoceptors are involved in regulation of lipolysis rate and microcirculation of adipose tissue. However, the observed changes in local blood flow were not related to glycerol output. PMID- 11907179 TI - Differential effects of sarcolemmal and mitochondrial K(ATP) channels activated by 17 beta-estradiol on reperfusion arrhythmias and infarct sizes in canine hearts. AB - We have demonstrated the effects of estrogen on modulation of ATP-sensitive K(+) channels; however, the subcellular location of these channels is unknown. The purpose of the present study was to investigate the role of the sarcolemmal and mitochondrial ATP-sensitive K(+) channels in a canine model of myocardial infarction after stimulation with 17 beta-estradiol. Anesthetized dogs were subjected to 60 min of the left anterior descending coronary artery occlusion followed by 3 h of reperfusion. Infarct size was markedly reduced in estradiol treated dogs compared with controls (14 +/- 6 versus 42 +/- 6%, P < 0.0001), indicating the effective dose of estradiol administrated. Pretreatment with the mitochondrial ATP-sensitive K(+) channel antagonist 5-hydroxydecanoate completely abolished estradiol-induced cardioprotection. The sarcolemmal ATP-sensitive K(+) channel antagonist 1-15-12-(5-chloro-o-anisamido)ethyl-methoxyphenyl)sulfonyl-3 methylthiourea (HMR 1098) did not significantly attenuate estradiol-induced infarct size limitation. In addition, estradiol administration significantly reduced the incidence and duration of reperfusion-induced ventricular tachycardia and ventricular fibrillation. Although 5-hydroxydecanoate alone caused no significant effect on the incidence of reperfusion arrhythmias in the presence or absence of estradiol, the administration of HMR 1098 abolished estrogen-induced improvement of reperfusion arrhythmias. Pretreatment with the estrogen-receptor antagonist faslodex (ICI 182,780) did not alter estrogen-induced infarct-limiting and antiarrhythmic effects. These results demonstrate that estrogen is cardioprotective against infarct sizes and fatal reperfusion arrhythmias by different ATP-sensitive K(+) channels for an estrogen receptor-independent mechanism. The infarct size-limiting and antiarrhythmic effects of estrogen were abolished by 5-hydroxydecanoate and HMR 1098, suggesting that the effects may result from activation of the mitochondrial and sarcolemmal ATP-sensitive K(+) channels, respectively. PMID- 11907180 TI - Pharmacological profile of a novel phosphodiesterase 4 inhibitor, 4-(8 benzo[1,2,5]oxadiazol-5-yl-[1,7]naphthyridin-6-yl)-benzoic acid (NVP-ABE171), a 1,7-naphthyridine derivative, with anti-inflammatory activities. AB - We investigated the pharmacology of a new class of phosphodiesterase 4 (PDE4) inhibitor, 6,8-disubstituted 1,7-naphthyridines, by using 4-(8 benzo[1,2,5]oxadiazol-5-yl-[1,7]naphthyridin-6-yl)-benzoic acid (NVP-ABE171) as a representative compound and compared its potency with the most advanced PDE4 inhibitor, undergoing clinical trials, Ariflo [cis-4-cyano-4-(3-cyclopentyloxy-4 methoxyphenyl-r-1-cyclohexanecarboxylic acid)]. NVP-ABE171 inhibited the activity of phosphodiesterase 4A, 4B, 4C, and 4D with respective IC(50) values of 602, 34, 1230, and 1.5 nM. Ariflo was about 40 times less potent. In human cells, NVP ABE171 inhibited the eosinophil and neutrophil oxidative burst, the release of cytokines by T cells, and the tumor necrosis factor-alpha release from monocytes, in the nanomolar range. Ariflo presented a similar inhibition profile but was 7 to 50 times less potent. In BALB/c mice challenged with lipopolysaccharide, NVP ABE171 inhibited the airway neutrophil influx and activation with an ED(50) in the range of 3 mg/kg. Ariflo was inactive up to a dose of 10 mg/kg. In ovalbumin sensitized Brown Norway rats, NVP-ABE171 inhibited the lipopolysaccharide-induced airway neutrophil influx and activation (ED(50) of 0.2 mg/kg) and the ovalbumin induced airway eosinophil influx and activation (ED(50) of 0.1 mg/kg). Ariflo was about 100 times less potent in both models. In the ovalbumin model, NVP-ABE171 had a duration of action of more than 24 h. NVP-ABE171 is a novel PDE4 inhibitor that shows activity both in vitro on human inflammatory cells and in vivo in animal models of lung inflammation. This compound class may have potential for the treatment of airway inflammatory conditions such as asthma and chronic obstructive pulmonary diseases. PMID- 11907181 TI - Attenuation of scopolamine-induced and age-associated memory impairments by the sigma and 5-hydroxytryptamine(1A) receptor agonist OPC-14523 (1-[3-[4-(3 chlorophenyl)-1-piperazinyl]propyl]-5-methoxy-3,4-dihydro-2[1H]-quinolinone monomethanesulfonate). AB - Sigma and 5-HT(1A) receptor stimulation can increase acetylcholine (ACh) release in the brain. Because ACh release facilitates learning and memory, we evaluated the degree to which OPC-14523 (1-[3-[4-(3-chlorophenyl)-1-piperazinyl]propyl]-5 methoxy-3,4-dihydro-2[1H]-quinolinone monomethane sulfonate), a novel sigma and 5 HT(1A) receptor agonist, can augment ACh release and improve learning impairments in rats due to cholinergic- or age-related deficits. Single oral administration of OPC-14523 improved scopolamine-induced learning impairments in the passive avoidance task and memory impairment in the Morris water maze. The chronic oral administration of OPC-14523 attenuated age-associated impairments of learning acquisition in the water maze and in the conditioned active-avoidance response test. OPC-14523 did not alter basal locomotion or inhibit acetylcholinesterase (AChE) activity at concentrations up to 100 microM and, unlike AChE inhibitors, did not cause peripheral cholinomimetic responses. ACh release in the dorsal hippocampus of freely moving rats increased after oral delivery of OPC-14523 and after local delivery of OPC-14523 into the hippocampus. The increases in hippocampal ACh release were blocked by the sigma receptor antagonist NE-100 (N,N dipropyl-2-[4-methoxy-3-(2-phenylethoxy)-phenyl]-ethylamine). Thus, OPC-14523 improves scopolamine-induced and age-associated learning and memory impairments by enhancing ACh release, due to a stimulation of sigma and probably 5-HT(1A) receptors. Combined sigma/5-HT(1A) receptor agonism may be a novel approach to ameliorate cognitive disorders associated with age-associated cholinergic deficits. PMID- 11907182 TI - The selective phosphodiesterase 4 inhibitor RP 73-401 reduced matrix metalloproteinase 9 activity and transforming growth factor-beta release during acute lung injury in mice: the role of the balance between Tumor necrosis factor alpha and interleukin-10. AB - Matrix metalloproteinases (MMPs) and transforming growth factor (TGF)-beta are involved in airway remodeling associated with the inflammatory process. In this study, we investigated the effect of RP 73-401 (piclamilast), a selective phosphodiesterase-4 inhibitor, on MMP-9 activity and TGF-beta production in two murine models of acute inflammation. In the first model, the lipopolysaccharide (LPS)-induced increase in neutrophils, MMP-9 activity, and tumor necrosis factor (TNF)-alpha and TGF-beta release in bronchoalveolar lavage (BAL) was significantly reduced by RP 73-401 pretreatment. In contrast, the BAL interleukin (IL)-10 level was decreased by LPS but restored by RP 73-401. IL-10 administration in LPS-exposed mice elicited a significant reduction in BAL neutrophilia, MMP-9 activity, and TNF-alpha release but not in TGF-beta production. In the second model, RP 73-401 inhibited BAL neutrophils but not MMP 9 activity and TGF-beta production that were induced by intranasal TNF-alpha. We demonstrated that RP 73-401 might modulate the expression of airway remodeling associated mediators such as MMP-9 and TGF-beta and that this effect seemed to be at least partially mediated by the balance between TNF-alpha and IL-10. PMID- 11907183 TI - Intravenous buprenorphine self-administration by detoxified heroin abusers. AB - Several sources indicate that intravenously administered buprenorphine may have significant abuse liability in humans. The present study evaluated the reinforcing effects of intravenously administered buprenorphine (0, 2, and 8 mg) in detoxified heroin-dependent participants during a 7.5-week inpatient study. Participants (n = 6) were detoxified from heroin over a 1.5-week period immediately after admission. Testing subsequently occurred in three 2-week blocks. During the first week of each 2-week block, the reinforcing effects of buprenorphine were evaluated. Participants first received a dose of buprenorphine and $20 and then were given either the opportunity to self-administer the dose or $20 during choice sessions. During the second week of each 2-week block, the direct effects of heroin were measured to evaluate potential long-lasting antagonist effects of buprenorphine. Progressive ratio break-point values were significantly higher after 2 and 8 mg of buprenorphine compared with placebo. Correspondingly, several positive subjective ratings increased after administration of active buprenorphine relative to placebo. Although there were few differences in peak effects produced by 2 versus 8 mg of buprenorphine, the higher buprenorphine dose generally produced longer-lasting effects. Heroin also produced dose-related increases in several subjective effects. Peak ratings produced by heroin were generally higher than peak ratings produced by buprenorphine. There was little evidence of residual antagonism produced by buprenorphine. These results demonstrate that buprenorphine served as a reinforcer under these conditions, and that it may have abuse liability in nonopioid-dependent individuals who abuse heroin. PMID- 11907184 TI - In utero ethanol suppresses cerebellar activator protein-1 and nuclear factor kappa B transcriptional activation in a rat fetal alcohol syndrome model. AB - A model of fetal alcohol syndrome was used to investigate prenatal ethanol effects on cerebellar transcription factors. Pregnant Sprague-Dawley rats were divided into three treatment groups: ethanol-exposed (E), calorically matched pair-fed (PF), and freely fed ad libitum (AL) groups. Ethanol exposure was stopped 2 days before parturition. The DNA binding in neonatal cerebella of the redox-sensitive transcription factors nuclear factor-kappa B (NF-kappa B) and activator protein-1 (AP-1) were determined by electrophoretic mobility shift assays. On the first postnatal day (PD1), there was decreased activation of these transcription factors in the E group relative to the control groups. The PD1 transcriptional effects were reversed as the neonate underwent development without further ethanol exposure. Western blot studies showed no corresponding decreases in protein amounts of both AP-1 and NF-kappa B components on PD1. Postnatal glutathione levels and catalase activity, as measures of oxidative stress hypothesized to be a probable cause of the transcriptional effects, showed no statistically significant effects attributable to ethanol. Examination of prenatal cerebella on embryonic day 20 (EM20), a time during ethanol exposure, showed DNA-binding trends similar to those of PD1. EM20 Western blot studies showed decreases in the levels of the active form of glycogen synthase kinase-3 (GSK-3). GSK-3 inhibition was reversed by PD1. Blocking of GSK-3 activity with gestational dietary lithium diminished both AP-1 and NF-kappa B DNA binding. Thus, prenatal ethanol exposure has the effect of diminishing pro-survival transcriptional activation, an effect possibly mediated by changes in GSK-3 activity. PMID- 11907185 TI - Effects of (+/-)-4-[[2-(1-methyl-2-pyrrolidinyl)ethyl]thio]phenol hydrochloride (SIB-1553A), a selective ligand for nicotinic acetylcholine receptors, in tests of visual attention and distractibility in rats and monkeys. AB - Nicotine, a nonselective ligand for nicotinic acetylcholine receptors (nAChRs), has been shown to improve attention and reduce distractibility in humans. Although the numerous side effects induced by nicotine prevent its use as a therapeutic agent, it is hypothesized that subtype-selective nAChR ligands may offer a potential therapeutic benefit to humans with attention deficits. In this study, we evaluated the attention-enhancing properties of (+/-)-4-[[2-(1-methyl-2 pyrrolidinyl)ethyl]thio]phenol hydrochloride (SIB-1553A), a ligand selective for neuronal nAChRs with predominant activity at the human beta 4 subtype. SIB-1553A was evaluated in a test of attention (i.e., five-choice serial reaction time task or SRTT) and distractibility (i.e., delayed matching to sample task with distractor or DMTS-D) in adult rats and monkeys, respectively. SIB-1553A did not improve SRTT performance in normal rats, but reversed deficits induced by the N methyl-D-aspartate (NMDA) antagonist dizocilpine. In the DMTS-D, SIB-1553A improved accuracy across several doses at the short delay intervals, which were affected most by distracting stimuli in adult monkeys. Subsequent testing with optimal doses for each monkey was also associated with significant improvements in DMTS-D accuracy at short delays, indicating the reproducibility of the drug effect. In both species, SIB-1553A had no significant effects on latencies for sample or choice selection and was not associated with adverse effects at efficacious doses. Although it remains to be further demonstrated, SIB-1553A may act through combined nicotinic and non-nicotinic mechanisms. Collectively, the present data suggest that in specific conditions SIB-1553A may improve certain aspects of attentional function in young adult rats and nonhuman primates without adverse side effects. PMID- 11907186 TI - Human organic anion transporters and human organic cation transporters mediate renal transport of prostaglandins. AB - Prostaglandin E(2) (PGE(2)) and prostaglandin F(2 alpha) (PGF(2 alpha)) have been used for the induction of labor and the termination of pregnancy. Renal excretion is shown to be an important pathway for the elimination of PGE(2) and PGF(2 alpha). The purpose of this study was to elucidate the molecular mechanism of renal PGE(2) and PGF(2 alpha) transport using cells stably expressing human organic anion transporter (hOAT) 1, hOAT2, hOAT3, and hOAT4, and human organic cation transporter (hOCT) 1 and hOCT2. A time- and dose-dependent increase in PGE(2) and PGF(2 alpha) uptake was observed in cells expressing hOAT1, hOAT2, hOAT3, hOAT4, hOCT1, and hOCT2. The K(m) values of PGE(2) uptake by hOAT1, hOAT2, hOAT3, hOAT4, hOCT1, and hOCT2 were 970, 713, 345, 154, 657, and 28.9 nM, respectively, whereas those of PGF(2 alpha) uptake by hOAT1, hOAT3, hOAT4, hOCT1, and hOCT2 were 575, 1092, 692, 477, and 334 nM, respectively. PGE(2) and PGF(2 alpha) significantly inhibited organic anion uptake by hOATs and organic cation uptake by hOCTs. In conclusion, considering the localization of these transporters, the results suggest that PGE(2) and PGF(2 alpha) transport in the basolateral membrane of the proximal tubule is mediated by multiple pathways including hOAT1, hOAT2, hOAT3, and hOCT2, whereas that in the apical side is mediated by hOAT4. PMID- 11907187 TI - Inhibitors of pentose phosphate pathway cause vasodilation: involvement of voltage-gated potassium channels. AB - Cytosolic reducing cofactors, such as NADPH and NADH, are thought to regulate vascular smooth muscle ion channel activity and vascular tone. In this study, the effects of pentose phosphate pathway (PPP) inhibitors, 6-aminonicotinamide (6 AN), epiandrosterone (EPI), and dehydroepiandrosterone (DHEA), on vascular tone were studied in isolated perfused lungs and pulmonary artery (PA) and aortic rings from rats. In addition, effects of 6-AN on voltage-gated K(+) (K(v)) current in PA smooth muscle cells (SMCs) were also examined. Pretreatment of lungs with 6-AN and EPI reduced the pressor response to acute hypoxia and decreased tissue NADPH levels. 6-AN, EPI, and DHEA relaxed isolated PA and aortic rings precontracted with 30 mM KCl in a dose-dependent manner. The PPP inhibitor induced PA relaxations were reduced in PA rings precontracted with 80 mM KCl but not by pretreatment with nitro-L-arginine or endothelial removal. Pretreatment of PA rings with tetraethylammonium chloride or 4-aminopyridine caused rightward shifts of concentration-relaxation curves for 6-AN, EPI, and DHEA. In contrast, glybenclamide, charybdotoxin, or apamin did not inhibit the relaxant effects of 6 AN, EPI, and DHEA. 6-AN caused an increase in K(v) current in PASMC. These results indicate that reduction of NADPH by the PPP inhibitors causes vasodilation at least partly through opening of K(v) channels. PMID- 11907188 TI - Sigma(2)-receptor regulation of dopamine transporter via activation of protein kinase C. AB - The elucidation of the mechanisms underlying sigma(2)-receptor activation and signal transduction is crucial to the understanding of sigma(2)-receptor function. Previous studies in our laboratory have demonstrated sigma(2)-receptor mediated regulation of the dopamine transporter (DAT) as measured by amphetamine stimulated release of [(3)H]dopamine (DA) from both rat striatal slices and PC12 cells. The regulation of the DAT in the PC12 cell model was dependent upon activation of Ca(2+)/calmodulin-dependent kinase II. We have now studied the second messenger systems involved in sigma(2)-receptor-mediated regulation of amphetamine-stimulated [(3)H]DA release in rat striatal slices, including Ca(2+)/calmodulin-dependent kinase II, protein kinase C, and sources of calcium required for the enhancement of release produced by sigma(2)-receptor activation. The Ca(2+)/calmodulin-dependent kinase II inhibitors 1-[N,O-bis-(5 isoquionolinesulfonyl)]-N-methyl-L-tyrosyl-4-phenylpiperazine and N-[2-[[[3-(4' chlorophenyl)-2-propenyl]methylamino]methyl]phenyl]-N-(2-hydroxyethyl)-4'-methoxy benzenesulfonamide phosphate did not significantly affect the (+)-pentazocine mediated enhancement of amphetamine-stimulated [(3)H]DA release. However, we found that an inhibitor of protein kinase C, 3-[1-[3-(dimethylamino)propyl]-1H indol-3-yl)-1H-pyrrole-2,5-dione, blocks the (+)-pentazocine-mediated enhancement in rat striatal slices. The protein kinase C activator phorbol 12-myristate 13 acetate, but not the inactive isophorbol 4 alpha,9 alpha,12 alpha,13 alpha,20 pentahydroxytiglia-1,6-dien-3-one, enhanced the amphetamine-stimulated [(3)H]DA release comparable to the enhancement seen by (+)-pentazocine alone. Additionally, the L-type voltage-dependent calcium channel inhibitor nitrendipine or prior treatment with thapsigargin, but not the N-type voltage-dependent calcium channel omega-conotoxin MVIIA, attenuated the (+)-pentazocine-mediated enhancement. Together, these data suggest that activation of sigma(2)-receptors results in the regulation of DAT activity via a calcium- and protein kinase C dependent signaling mechanism. PMID- 11907189 TI - Kinetic interactions of dopamine and dobutamine with human catechol-O methyltransferase and monoamine oxidase in vitro. AB - Dopamine and dobutamine are often infused together into acutely ill patients requiring temporary support of cardiac and renal function, but whether these catecholamines affect the metabolic clearance of each other is not established. We determined the kinetics of dopamine and dobutamine as substrates and inhibitors of each other, i.e., apparent V(max), K(m), and K(i), with crude preparations of human blood mononuclear cell catechol-O-methyltransferase (COMT) and platelet monoamine oxidase (MAO) at pH 7.4 and 37 degrees C. Values of V(max) for dopamine and dobutamine as substrates for COMT were 0.45 and 0.59 nmol of 3-O methyl product formed per milligram of protein per minute, whereas those for K(m) were 0.44 and 0.05 mM, respectively. Dopamine and dobutamine were competitive inhibitors of each other in this reaction. The K(i) for dopamine as an inhibitor of dobutamine methylation was 1.5 mM, whereas that for dobutamine as an inhibitor of dopamine methylation was 0.015 mM. Dopamine but not dobutamine was a substrate for MAO. The V(max) for dihydroxyphenylacetaldehyde formation from dopamine was 0.29 nmol/mg protein/min and the K(m) for dopamine was 0.38 mM. Dobutamine was a noncompetitive inhibitor of dopamine oxidation in this reaction (K(i) congruent with 1.19 mM). The high apparent K(m) and K(i) values derived for dopamine and dobutamine when tested with these two human enzymes in vitro suggest that these catecholamines do not interfere with the metabolism of each other when both are infused together at therapeutic concentrations. PMID- 11907190 TI - 4-(2-Chloro-4-methoxy-5-methylphenyl)-N-[(1S)-2-cyclopropyl-1-(3-fluoro-4 methylphenyl)ethyl]5-methyl-N-(2-propynyl)-1,3-thiazol-2-amine hydrochloride (SSR125543A): a potent and selective corticotrophin-releasing factor(1) receptor antagonist. I. Biochemical and pharmacological characterization. AB - 4-(2-Chloro-4-methoxy-5-methylphenyl)-N-[(1S)-2-cyclopropyl-1- (3-fluoro-4 methylphenyl)ethyl]5-methyl-N-(2-propynyl)-1,3-thiazol-2-amine hydrochloride (SSR125543A), a new 2-aminothiazole derivative, shows nanomolar affinity for human cloned or native corticotrophin-releasing factor (CRF)(1) receptors (pK(i) values of 8.73 and 9.08, respectively), and a 1000-fold selectivity for CRF(1) versus CRF(2 alpha) receptor and CRF binding protein. SSR125543A antagonizes CRF induced stimulation of cAMP synthesis in human retinoblastoma Y 79 cells (IC(50) = 3.0 +/- 0.4 nM) and adrenocorticotropin hormone (ACTH) secretion in mouse pituitary tumor AtT-20 cells. SSR125543A is devoid of agonist activity in these models. Its brain penetration was demonstrated in rats by using an ex vivo [(125)I-Tyr(0)] ovine CRF binding assay. SSR125543A displaced radioligand binding to the CRF(1) receptor in the brain with an ID(50) of 6.5 mg/kg p.o. (duration of action >24 h). SSR125543A also inhibited the increase in plasma ACTH levels elicited in rats by i.v. CRF (4 microg/kg) injection (ID(50) = 1, 5, or 5 mg/kg i.v., i.p., and p.o., respectively); this effect lasted for more than 6 h when the drug was given orally at a dose of 30 mg/kg. SSR125543A (10 mg/kg p.o.) reduced by 73% the increase in plasma ACTH levels elicited by a 15-min restraint stress in rats. Moreover, SSR125543A (20 mg/kg i.p.) also antagonized the increase of hippocampal acetylcholine release induced by i.c.v. injection of 1 microg of CRF in rats. Finally, SSR125543A reduced forepaw treading induced by i.c.v. injection of 1 microg of CRF in gerbils (ID(50) = approximately 10 mg/kg p.o.). Altogether, these data indicate that SSR125543A is a potent, selective, and orally active CRF(1) receptor antagonist. PMID- 11907191 TI - 4-(2-Chloro-4-methoxy-5-methylphenyl)-N-[(1S)-2-cyclopropyl-1-(3-fluoro-4 methylphenyl)ethyl]5-methyl-N-(2-propynyl)-1, 3-thiazol-2-amine hydrochloride (SSR125543A), a potent and selective corticotrophin-releasing factor(1) receptor antagonist. II. Characterization in rodent models of stress-related disorders. AB - The present study investigated the effects of the novel corticotrophin-releasing factor (CRF)(1) receptor antagonist 4-(2-chloro-4-methoxy-5-methylphenyl)-N-[(1S) 2-cyclopropyl-1-(3-fluoro-4-methylphenyl)ethyl]5-methyl-N-(2-propynyl)-1,3 thiazol-2-amine hydrochloride (SSR125543A) in a variety of rodent models of anxiety, including conflict procedures (punished drinking and four-plate), exploration models (elevated plus-maze and light/dark), a fear/anxiety defense test battery, and several procedures based on stress-induced changes in physiological (isolation-induced hyperthermia and tail pinch-induced cortical norepinephrine release) or behavioral (social defeat-induced anxiety, maternal separation-induced vocalization) parameters. Moreover, the effects of SSR125543A were investigated in acute (forced swimming) and chronic (chronic mild stress; CMS) models of depression. SSR125543A and the CRF(1) receptor antagonist antalarmin displayed limited efficacy in exploration-based anxiety models. In contrast, both compounds produced clear-cut anxiolytic-like activity in models involving inescapable stress, including the conflict procedures, the social defeat-induced anxiety paradigm and the defense test battery (3-30 mg/kg i.p. or p.o.). These effects paralleled those of the anxiolytic diazepam. In addition, SSR125543A and antalarmin antagonized stress-induced hyperthermia, distress vocalization, and cortical norepinephrine release. In the forced swimming test, 30 mg/kg p.o. SSR125543A and 3 to 30 mg/kg p.o. antalarmin produced clear antidepressant-like effects. These latter results were strengthened by the findings from the CMS, which showed that repeated administration of 10 mg/kg i.p. SSR125543A for 30 days improved the degradation of the physical state, the reduction of body weight gain, and anxiety produced by stress. Together, these data indicate that SSR125543A shows good activity in acute and chronic tests of unavoidable stress exposure, suggesting that it may have a potential in the treatment of depression and some forms of anxiety disorders. PMID- 11907192 TI - Cyclosporine adversely affects baroreflexes via inhibition of testosterone modulation of cardiac vagal control. AB - Previous studies have shown that the immunosuppressant drug cyclosporine A attenuates arterial baroreceptor function. This study investigated whether the modulatory effect of cyclosporine on baroreceptor function involves inhibition of the baroreflex-facilitatory effect of testosterone. The role of cardiac autonomic control in cyclosporine-testosterone baroreflex interaction was also investigated. Baroreflex curves relating bradycardic responses to increments in blood pressure evoked by phenylephrine were constructed in conscious, sham operated, castrated rats and in testosterone-replaced castrated (CAS + T) rats in the absence and presence of cyclosporine. The slopes of the curves were taken as an index of the baroreflex sensitivity (BRS). Short-term (11-13 days) cyclosporine treatment or castration reduced plasma testosterone levels and caused similar attenuation of the reflex bradycardia, as indicated by the significantly smaller BRS compared with sham-operated values (-0.97 +/- 0.07, 0.86 +/- 0.06, and -1.47 +/- 0.10 beats/min/mm Hg, respectively). The notion that androgens facilitate baroreflexes is further confirmed by the observation that testosterone replacement of castrated rats restored plasma testosterone and BRS to sham-operated levels. Cyclosporine had no effect on BRS in castrated rats but caused a significant reduction in CAS + T rats. Muscarinic blockade by atropine caused approximately 60% reduction in the BRS in sham-operated rats, an effect that was significantly and similarly diminished by castration, cyclosporine, or their combination. beta-Adrenergic blockade by propranolol caused no significant changes in BRS. These findings suggest that cyclosporine attenuates baroreflex responsiveness via, at least partly, inhibition of the testosterone-induced facilitation of cardiomotor vagal control. PMID- 11907193 TI - Activation of endogenous nitric oxide synthase coupled with methacholine-induced exocytosis in rat parotid acinar cells. AB - Methacholine (MCh) interacted with M(3) muscarinic receptors in rat parotid tissue slices and induced amylase secretion. MCh- and calcimycin-induced exocytosis was completely inhibited by N-[2-(N-(4-chlorocinnamyl)-N methylaminomethyl)phenyl]-N-[2-hydroxyethyl]-4-methoxybenzenesulfonamide, N(G) nitro-L-arginine methylester (L-NAME), 1H-(1,2,4)-oxadiazolo[4,3-a]quinoxaline-1 one, and 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide, suggesting that activations of calmodulin (CaM) kinase II, nitric oxide synthase (NOS), and cGMP-dependent protein kinase (PKG) were coupled with the exocytosis. These suggestions were supported by the results that exposure of the slices to MCh induced a rapid increase in these enzyme activities. Western blot analysis showed that neuronal NOS (nNOS) was expressed in isolated parotid acinar cells of rats. To measure nitric oxide (NO) production in response to the stimulation with MCh in real time, the isolated parotid acinar cells had been preloaded with 4,5 diaminofluorescein diacetate and incubated with the agonist. MCh (1 microM) induced a fast increase in 4,5-diaminofluorescein fluorescence, corresponding to an increase in the NO synthesis in the presence of extracellular Ca(2+) but not in the absence of it. When the isolated parotid acinar cells preloaded with L NAME or 2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethylester) were treated simultaneously with MCh, the increase in the fluorescence also was not observed. The MCh-induced increase in the fluorescence was not observed in the cells incubated in the absence of extracellular calcium, showing the importance of Ca(2+) entry from extracellular sites for MCh-induced NOS activation. These results indicate that nNOS is endogenously present in rat parotid acinar cells and that the rapid activation of this enzyme together with those of CaM kinase II and PKG contributes to MCh-induced amylase secretion. PMID- 11907194 TI - Characterization of mu, kappa, and delta opioid binding in amphibian whole brain tissue homogenates. AB - Opioid agonists produce analgesia in mammals through the activation of mu, kappa, or delta opioid receptors. Previous behavioral and binding studies from our laboratory using an amphibian model suggested that mu, kappa, or delta opioid agonists may activate a single type of opioid receptor in the grass frog, Rana pipiens. In the present study, kinetic, saturation, and competitive binding profiles for three opioid radioligands, [(3)H]DAMGO ([D-Ala(2),N-Me-Phe(4),Gly(5) ol]-enkephalin) (mu-selective), [(3)H]U65953 [(5 alpha, 7 alpha,8 beta)-(+)-N methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro[4.5]dec-8-yl]-benzeneacetamide] (kappa selective), and [(3)H]DPDPE ([D-Pen(2),D-Pen(5)]-enkephalin) (delta-selective) were determined using frog whole brain homogenates. Kinetic analyses and experimentally derived values from saturation experiments gave affinity constants (K(D)) in the low nanomolar range. The density of opioid binding sites (B(max)) was 224.4, 118.6, and 268.9 fmol/mg for mu, kappa, and delta opioid radioligands, respectively. The affinity values did not significantly differ among the three opioid radioligands, but the kappa radioligand bound to significantly fewer sites than did the mu or delta radioligands. K(i) values for unlabeled mu, kappa, and delta competitors, including highly selective opioid antagonists, were consistent with each radioligand selectivity profile. The present data suggest that mu, kappa, and delta opioid radioligands bind to distinct opioid receptors in amphibians that are surprisingly similar to those found in mammalian brain. PMID- 11907195 TI - Alterations in responses of ventral pallidal neurons to excitatory amino acids after long-term dopamine depletion. AB - The present study explored the possibility that excitatory amino acid (EAA) sensitivity within the ventral pallidum (VP) is altered by long-term removal of dopamine (DA). Electrophysiological experiments were conducted in chloral hydrate anesthetized rats 21 to 28 days after they received unilateral substantia nigra injections of the dopaminergic toxin 6-hydroxydopamine (6-OHDA). VP neurons increased firing at low microiontophoretic ejection currents of the EAA agonists N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA); however, high currents decreased action potential amplitude and rapidly caused cessation of neuronal firing. These responses likely reflected the induction of depolarization block for they were reversed by coiontophoresis of the hyperpolarizing transmitter gamma-aminobutyric acid (GABA) at ejection current levels that normally suppressed firing. The ability of NMDA and AMPA to induce such inactivation was greater in the VP of 6-OHDA-lesioned hemispheres, but unchanged in reserpinized rats, verifying that the alterations in responding to NMDA were the result of chronic, rather than acute, DA removal. The adaptations do not appear to be the consequence of a diminished GABAergic tone for the ability of bicuculline to increase firing (due to blocking a tonic GABAergic input) was not changed. However, low ejection currents of GABA that were insufficient to alter firing rate greatly attenuated the ability of NMDA to induce an apparent depolarization inactivation when coiontophoresed with NMDA onto VP neurons of the lesioned, but not the unlesioned, hemisphere. These studies show that chronic DA removal altered the EAA-induced amplitude-decreasing (i.e., the apparent depolarization inactivation) effects in VP neurons in the absence of a decrease in GABAergic tone. PMID- 11907196 TI - In vitro analysis of human drug glucuronidation and prediction of in vivo metabolic clearance. AB - The glucuronidation of a number of commonly used hepatic uridine diphosphate glucuronosyltransferase drug substrates has been studied in human tissue microsomes. Prediction of in vivo hepatic drug glucuronidation from liver microsomal data yielded a consistent 10-fold under-prediction. Consideration of protein binding was observed to be pivotal when predicting in vivo glucuronidation for acid substrates. Studies using human intestinal microsomes demonstrated the majority of drugs to be extensively glucuronidated such that the intrinsic clearance (CL(int)) of ethinylestradiol (CL(int) = 1.3 microl/min/mg) was twice that obtained using human liver microsomes (CL(int) = 0.7 microl/min/mg). The potential extrahepatic in vivo glucuronidation was calculated for a range of drug substrates from human microsomal data. These results indicate the contribution of intestinal drug glucuronidation to systemic drug clearance to be much less than either hepatic or renal glucuronidation. Therefore, data obtained with intestinal microsomes may be misleading in the assessment of the contribution of this organ to systemic glucuronidation. The use of hepatocytes to assess metabolic stability for drugs predominantly metabolized by glucuronidation was also investigated. Metabolic clearances for a range of drugs obtained using fresh preparations of human hepatocytes predicted accurately hepatic clearance reported in vivo. The use of cryopreserved hepatocytes as an in vitro tool to predict in vivo metabolism was also assessed with an excellent correlation obtained for a number of extensively glucuronidated drugs (R(2) = 0.80, p < 0.001). PMID- 11907197 TI - Intranasal delivery of morphine. AB - Morphine administered nasally to humans as a simple solution is only absorbed to a limited degree, with a bioavailability of the order of 10% compared with intravenous administration. This article describes the development of novel nasal morphine formulations based on chitosan, which, in the sheep model, provide a highly increased absorption with a 5- to 6-fold increase in bioavailability over simple morphine solutions. The chitosan-morphine nasal formulations have been tested in healthy volunteers in comparison with a slow i.v. infusion (over 30 min) of morphine. The results show that the nasal formulation was rapidly absorbed with a T(max) of 15 min or less and a bioavailability of nearly 60%. The shape of the plasma profile for nasal delivery of the chitosan-morphine formulation was similar to the one obtained for the slow i.v. administration of morphine. Furthermore, the metabolite profile obtained after the nasal administration of the chitosan-morphine nasal formulation was essentially identical to the one obtained for morphine administered by the intravenous route. The levels of both morphine-6-glucuronide and morphine-3-glucuronide were only about 25% of that found after oral administration of morphine. It is concluded that a properly designed nasal morphine formulation (such as one with chitosan) can result in a non-injectable opioid product capable of offering patients rapid and efficient pain relief. PMID- 11907198 TI - Simian immunodeficiency virus Nef protein delays the progression of CD4+ T cells through G1/S phase of the cell cycle. AB - Human and simian immunodeficiency virus (HIV and SIV, respectively) infections are characterized by gradual depletion of CD4+ T cells. The underlying mechanisms of CD4+ T-cell depletion and HIV and SIV persistence are not fully determined. The Nef protein is expressed early in infection and is necessary for pathogenesis. Nef can cause T-cell activation and downmodulates cell surface signaling molecules. However, the effect of Nef on the cell cycle has not been well characterized. To determine the role of Nef in the cell cycle, we investigated whether the SIV Nef protein can modulate cell proliferation and apoptosis in CD4+ Jurkat T cells. We developed a CD4+ Jurkat T-cell line that stably expresses SIV Nef under the control of an inducible promoter. Alterations in cell proliferation were determined by flow cytometry using stable intracytoplasmic fluorescent dye 5- and 6-carboxyfluorescein diacetate succinimidyl ester and bromodeoxyuridine incorporation. Apoptotic cell death was measured by annexin V and propidium iodide staining. Our results demonstrated that SIV Nef inhibited Fas-induced apoptosis in these cells and that the mechanism involved upregulation of the Bcl-2 protein. SIV Nef suppressed CD4+ T cell proliferation by inhibiting the progression of cells into S phase of the cell cycle. Suppression involved an upregulation of cyclin-dependent kinase inhibitors p21 and p27 and the downregulation of cyclin D1 and cyclin A. In summary, inhibition of apoptosis by Nef can lead to persistence of infected cells and can support viral replication. In addition, a Nef-mediated delay in cell cycle progression may contribute to CD4+ T-cell anergy/depletion seen in HIV and SIV disease. PMID- 11907199 TI - Antiviral effects of an iminosugar derivative on flavivirus infections. AB - Endoplasmic reticulum (ER) alpha-glucosidase inhibitors, which block the trimming step of N-linked glycosylation, have been shown to eliminate the production of several ER-budding viruses. Here we investigated the effects of one such inhibitor, N-nonyl-deoxynojirimycin (NN-DNJ), a 9-carbon alkyl iminosugar derivative, on infection by Japanese encephalitis virus (JEV) and dengue virus serotype 2 (DEN-2). In the presence of NN-DNJ, JEV and DEN-2 infections were suppressed in a dose-dependent manner. This inhibitory effect appeared to influence DEN-2 infection more than JEV infection, since lower concentrations of NN-DNJ substantially blocked DEN-2 replication. Secretion of the flaviviral glycoproteins E and NS1 was greatly reduced, and levels of DEN-2 viral RNA replication measured by fluorogenic reverse transcription-PCR were also decreased, by NN-DNJ. Notably, the viral glycoproteins, prM, E, and NS1 were found to associate transiently with the ER chaperone calnexin, and this interaction was affected by NN-DNJ, suggesting a potential role of calnexin in the folding of flaviviral glycoproteins. Additionally, in a mouse model of lethal challenge by JEV infection, oral delivery of NN-DNJ reduced the mortality rate. These findings show that NN-DNJ has an antiviral effect on flavivirus infection, likely through interference with virus replication at the posttranslational modification level, occurring mainly in the ER. PMID- 11907200 TI - Replication fidelity of the supF gene integrated in the thymidine kinase locus of herpes simplex virus type 1. AB - Recombinant viruses were constructed to have an Escherichia coli replicon containing a mutagenesis marker, the supF gene, integrated within the thymidine kinase locus (tk) of herpes simplex virus type 1. These viruses expressed either wild-type or mutant DNA polymerase (Pol) and were tested in a mutagenesis assay for the fidelity of their replication of the supF gene. A mutation frequency of approximately 10(-4) was observed for wild-type strain KOS-derived recombinants in their replication of the supF gene. However, recombinants derived from the PAA(r)5 Pol mutant, which has been demonstrated to have an antimutator phenotype in replicating the tk gene, had three- to fourfold increases in supF mutation frequency (P < 0.01), a result similar to that exhibited when the supF gene was induced to replicate as episomal DNA (Y. T. Hwang, B.-Y. Liu, C.-Y. Hong, E. J. Shillitoe, and C. B. C. Hwang, J. Virol. 73:5326-5332, 1999). Thus, the PAA(r)5 Pol mutant had an antimutator function in replicating the tk gene and was less accurate in replicating the supF gene than was the wild-type strain. The spectra of mutations and distributions of substituted bases within the supF genes that replicated as genomic DNA were different from those in the genes that replicated as episomal DNA. Therefore, the differences in sequence contents between the two target genes influenced the accuracy of the Pol during viral replication. Furthermore, the replication mode of the target gene also affected the mutational spectrum. PMID- 11907202 TI - Feline immunodeficiency virus xenoinfection: the role of chemokine receptors and envelope diversity. AB - The use of chemokine receptors as cell recognition signals is a property common to several lentiviruses, including feline, human, and simian immunodeficiency viruses. Previously, two feline immunodeficiency virus (FIV) isolates, V1CSF and Petaluma, were shown to use chemokine receptors in a strain-dependent manner to infect human peripheral blood mononuclear cells (PBMC) (J. Johnston and C. Power, J. Virol. 73:2491-2498, 1999). Since the sequences of these viruses differed primarily in regions of the FIV envelope gene implicated in receptor use and cell tropism, envelope chimeras of V1CSF and Petaluma were constructed to investigate the role of envelope diversity in the profiles of chemokine receptors used by FIV to infect primate cells. By use of a receptor-blocking assay, all viruses were found to infect human and macaque PBMC through a mechanism involving the CXCR4 receptor. However, infection by viruses encoding the V3-to-V5 region of the V1CSF surface unit was also inhibited by blockade of the CCR3 or CCR5 receptor. Similar results were obtained with GHOST cells, human osteosarcoma cells expressing specific combinations of chemokine receptors. CXCR4 was required for infection by all FIV strains, but viruses expressing the V3-to-V5 region of V1CSF required the concurrent presence of either CCR3 or CCR5. In contrast, CXCR4 alone was sufficient to allow infection of GHOST cells by FIV strains possessing the V3-to V5 region of Petaluma. To assess the role of primate chemokine receptors in productive infection, Crandell feline kidney (CrFK) cells that expressed human CXCR4, CCR3, or CCR5 in addition to feline CXCR4 were generated. Sustained infection by viruses encoding the V3-to-V5 region of V1CSF was detected in CrFK cells expressing human CCR3 or CCR5 but not in cells expressing CXCR4 alone, while all CrFK cell lines were permissive to viruses encoding the V3-to-V5 region of Petaluma. These results indicate that FIV uses chemokine receptors to infect both human and nonhuman primate cells and that the profiles of these receptors are dependent on envelope sequence, and they provide insights into the mechanism by which xenoinfections may occur. PMID- 11907201 TI - Therapeutic immunization with a virion host shutoff-defective, replication incompetent herpes simplex virus type 1 strain limits recurrent herpetic ocular infection. AB - Immunization of mice with herpes simplex virus type 1 (HSV-1) mutant viruses containing deletions in the gene for virion host shutoff (vhs) protein diminishes primary and recurrent corneal infection with wild-type HSV-1. vhs mutant viruses are severely attenuated in vivo but establish latent infections in sensory neurons. A safer HSV-1 mutant vaccine strain, Delta41Delta29, has combined vhs and replication (ICP8-) deficits and protects BALB/c mice against primary corneal infection equivalent to a vhs- strain (BGS41). Here, we tested the hypothesis that Delta41Delta29 can protect as well as BGS41 in a therapeutic setting. Because immune response induction varies with the mouse and virus strains studied, we first determined the effect of prophylactic Delta41Delta29 vaccination on primary ocular infection of NIH inbred mice with HSV-1 McKrae, a model system used to evaluate therapeutic vaccines. In a dose-dependent fashion, prophylactic Delta41Delta29 vaccination decreased postchallenge tear film virus titers and ocular disease incidence and severity while eliciting high levels of HSV-specific antibodies. Adoptive transfer studies demonstrated a dominant role for immune serum and a lesser role for immune cells in mediating prophylactic protection. Therapeutically, vaccination with Delta41Delta29 effectively reduced the incidence of UV-B-induced recurrent virus shedding in latently infected mice. Therapeutic Delta41Delta29 and BGS41 vaccination decreased corneal opacity and delayed-type hypersensitivity responses while elevating antibody titers, compared to controls. These data indicate that replication is not a prerequisite for generation of therapeutic immunity by live HSV mutant virus vaccines and raise the possibility that genetically tailored replication-defective viruses may make effective and safe therapeutic vaccines. PMID- 11907203 TI - The Tat protein of human immunodeficiency virus type 1 (HIV-1) can promote placement of tRNA primer onto viral RNA and suppress later DNA polymerization in HIV-1 reverse transcription. AB - Human immunodeficiency virus type-1 Tat has been proposed to play a role in the regulation of reverse transcription. We previously demonstrated that wild-type Tat can augment viral infectivity by suppressing the reverse transcriptase (RT) reaction at late stages of the viral life cycle in order to prevent the premature synthesis of potentially deleterious viral DNA products. Here we have performed a detailed analysis of the cell-free reverse transcription reaction to elucidate the mechanism(s) whereby Tat can affect this process. Our results show that Tat can suppress nonspecific DNA elongation while moderately affecting the specific initiation stage of reverse transcription. In addition, Tat has an RNA-annealing activity and can promote the placement of tRNA onto viral RNA. This points to a functional homology between Tat and the viral nucleocapsid (NC) protein that is known to be directly involved in this process. Experiments using a series of mutant Tat proteins revealed that the cysteine-rich and core domains of Tat are responsible for suppression of DNA elongation, while each of the cysteine-rich, core, and basic domains, as well as a glutamine-rich region in the C-terminal domain, are important for the placement of tRNA onto the viral RNA genome. These results suggest that Tat can play at least two different roles in the RT reaction, i.e., suppression of DNA polymerization and placement of tRNA onto viral RNA. We believe that the first of these activities of Tat may contribute to the overall efficiency of reverse transcription of the viral genome during a new round of infection as well as to enhanced production of infectious viral particles. We hypothesize that the second activity, illustrating functional homology between Tat and NC, suggests a potential role for NC in the displacement of Tat during viral maturation. PMID- 11907204 TI - Decreased frequency of cytomegalovirus (CMV)-specific CD4+ T lymphocytes in simian immunodeficiency virus-infected rhesus macaques: inverse relationship with CMV viremia. AB - The frequency of cytomegalovirus (CMV)-specific CD4+ T lymphocytes was determined in CMV-seropositive rhesus macaques with or without simian immunodeficiency virus (SIV) infection by using the sensitive assays of intracellular cytokine staining and gamma interferon ELISPOT. Both techniques yielded 3- to 1,000-fold-higher frequencies of CMV-specific CD4+ T lymphocytes than traditional proliferative limiting dilution assays. The median frequency of CMV-specific CD4+ T lymphocytes in 23 CMV-seropositive SIV-negative macaques was 0.63% (range, 0.16 to 5.8%). The majority of CMV-specific CD4+ T lymphocytes were CD95(pos) and CD27(lo) but expressed variable levels of CD45RA. A significant reduction (P < 0.05) in the frequency of CMV-specific CD4+ T lymphocytes was observed in pathogenic SIV infected macaques but not in macaques infected with live attenuated strains of SIV. CMV-specific CD4+ T lymphocytes were not detected in six of nine pathogenic SIV-infected rhesus macaques. CMV DNA was detected in the plasma of four of six of these macaques but in no animal with detectable CMV-specific CD4+ T lymphocytes. In pathogenic SIV-infected macaques, loss of CMV-specific CD4+ T lymphocytes was not predicted by the severity of CD4+ T lymphocytopenia. Neither was it predicted by the pre-SIV infection frequencies of CD45RA(neg) or CCR5(pos) CMV-specific CD4+ T lymphocytes. However, the magnitude of activation, as evidenced by the intensity of CD40L expression on CMV-specific CD4+ T lymphocytes pre-SIV infection, was three- to sevenfold greater in the two macaques that subsequently lost these cells after SIV infection than in the two macaques that retained CMV-specific CD4+ T lymphocytes post-SIV infection. Future longitudinal studies with these techniques will facilitate the study of CMV pathogenesis in AIDS. PMID- 11907206 TI - Rapid construction of adenoviral vectors by lambda phage genetics. AB - Continued improvements of adenoviral vectors require the investigation of novel genome configurations. Since adenovirus can be generated directly by transfecting packaging cell lines with viral genomes isolated from plasmid DNA, it is possible to separate genome construction from virus production. In this way failure to generate a virus is not associated with an inability to generate the desired genome. We have developed a novel lambda-based system that allows rapid modification of the viral genome by double homologous recombination in Escherichia coli. The recombination reaction and newly generated genome may reside in a recombination-deficient bacterial host for enhanced plasmid stability. Furthermore, the process is independent of any restriction endonucleases. The strategy relies on four main steps: (i) homologous recombination between an adenovirus cosmid and a donor plasmid (the donor plasmid carries the desired modification[s] and flanking regions of homology to direct its recombination into the viral genome); (ii) in vivo packaging of the recombinant adenoviral cosmids during a productive lambda infection; (iii) transducing a recombination-deficient E. coli lambda lysogen with the generated lysate (the lysogen inhibits the helper phage used to package the recombinant andenoviral cosmid from productively infecting and destroying the host bacteria); (iv) effectively selecting for the desired double-recombinant cosmid. Approximately 10,000 double-recombinant cosmids are recovered per reaction with essentially all of them being the correct double-recombinant molecule. This system was used to generate quickly and efficiently adenoviral genomes deficient in the E1/E3 and E1/E3/E4 regions. The basis of this technology allows any region of the viral genome to be readily modified for investigation of novel configurations. PMID- 11907205 TI - Recognition of the measles virus nucleocapsid as a mechanism of IRF-3 activation. AB - The mechanisms of cellular recognition for virus infection remain poorly understood despite the wealth of information regarding the signaling events and transcriptional responses that ensue. Host cells respond to viral infection through the activation of multiple signaling cascades, including the activation of NF-kappaB, c-Jun/ATF-2 (AP-1), and the interferon regulatory factors (IRFs). Although viral products such as double-stranded RNA (dsRNA) and the processes of viral binding and fusion have been implicated in the activation of NF-kappaB and AP-1, the mechanism(s) of IRF-1, IRF-3, and IRF-7 activation has yet to be fully elucidated. Using recombinant measles virus (MeV) constructs, we now demonstrate that phosphorylation-dependent IRF-3 activation represents a novel cellular detection system that recognizes the MeV nucleocapsid structure. At low multiplicities of infection, IRF-3 activation is dependent on viral transcription, since UV cross-linking and a deficient MeV containing a truncated polymerase L gene failed to induce IRF-3 phosphorylation. Expression of the MeV nucleocapsid (N) protein, without the requirement for any additional viral proteins or the generation of dsRNA, was sufficient for IRF-3 activation. In addition, the nucleocapsid protein was found to associate with both IRF-3 and the IRF-3 virus-activated kinase, suggesting that it may aid in the colocalization of the kinase and the substrate. Altogether, this study suggests that IRF-3 recognizes nucleocapsid structures during the course of an MeV infection and triggers the induction of interferon production. PMID- 11907207 TI - Herpes simplex virus vectors elicit durable immune responses in the presence of preexisting host immunity. AB - Herpes simplex virus (HSV) recombinants are being developed as vaccine vectors for the expression of heterologous antigens. There is concern, however, that preexisting HSV immunity may decrease their effectiveness. We have addressed this issue in an animal model. Immunized mice were inoculated with a replication defective HSV-1 vector that expressed the Escherichia coli beta-galactosidase protein as a model antigen. We assessed vector efficacy by analyzing the immunoglobulin G (IgG) antibody response and cellular proliferative response directed against beta-galactosidase. We report that the ability of the vector to induce antibody or proliferative responses was not diminished by preexisting immunity to HSV. Of further note, the anti-HSV and anti-beta-galactosidase IgG responses following vector administration were extremely durable in both immunized and naive mice. These results indicate that the ability of a replication-defective HSV-derived vaccine vector to elicit long-lived immune responses in mice is not impaired by prior HSV exposure. PMID- 11907208 TI - Selection of RNA aptamers that are specific and high-affinity ligands of the hepatitis C virus RNA-dependent RNA polymerase. AB - In order to find small RNA molecules that are specific and high-affinity ligands of nonstructural 5B (NS5B) polymerase, we screened by SELEX (systematic evolution of ligands by exponential amplification) a structurally constrained RNA library with an NS5BDeltaC55 enzyme carrying a C-terminal biotinylation sequence. Among the selected clones, two aptamers appeared to be high-affinity ligands of NS5B, with apparent dissociation constants in the low nanomolar range. They share a sequence that can assume a stem-loop structure. By mutation analysis, this structure has been shown to correspond to the RNA motif responsible for the tight interaction with NS5B. The aptamers appeared to be highly specific for the hepatitis C virus (HCV) polymerase since interaction with the GB virus B (GBV-B) NS5B protein cannot be observed. This is consistent with the observation that the activity of the HCV NS5B polymerase is efficiently inhibited by the selected aptamers, while neither GBV-B nor poliovirus 3D polymerases are affected. The mechanism of inhibition of the NS5B activity turned out to be noncompetitive with respect to template RNA, suggesting that aptamers and template RNA do not bind to the same site. As a matter of fact, mutations introduced in a basic exposed surface of the thumb domain severely impaired both the binding of and activity inhibition by the RNA aptamers. PMID- 11907209 TI - RNA replication of mouse hepatitis virus takes place at double-membrane vesicles. AB - The replication complexes (RCs) of positive-stranded RNA viruses are intimately associated with cellular membranes. To investigate membrane alterations and to characterize the RC of mouse hepatitis virus (MHV), we performed biochemical and ultrastructural studies using MHV-infected cells. Biochemical fractionation showed that all 10 of the MHV gene 1 polyprotein products examined pelleted with the membrane fraction, consistent with membrane association of the RC. Furthermore, MHV gene 1 products p290, p210, and p150 and the p150 cleavage product membrane protein 1 (MP1, also called p44) were resistant to extraction with Triton X-114, indicating that they are integral membrane proteins. The ultrastructural analysis revealed double-membrane vesicles (DMVs) in the cytoplasm of MHV-infected cells. The DMVs were found either as separate entities or as small clusters of vesicles. To determine whether MHV proteins and viral RNA were associated with the DMVs, we performed immunocytochemistry electron microscopy (IEM). We found that the DMVs were labeled using an antiserum directed against proteins derived from open reading frame 1a of MHV. By electron microscopy in situ hybridization (ISH) using MHV-specific RNA probes, DMVs were highly labeled for both gene 1 and gene 7 sequences. By combined ISH and IEM, positive-stranded RNA and viral proteins localized to the same DMVs. Finally, viral RNA synthesis was detected by labeling with 5-bromouridine 5'-triphosphate. Newly synthesized viral RNA was found to be associated with the DMVs. We conclude from these data that the DMVs carry the MHV RNA replication complex and are the site of MHV RNA synthesis. PMID- 11907210 TI - Parameters of human hepatitis delta virus genome replication: the quantity, quality, and intracellular distribution of viral proteins and RNA. AB - Assembly of hepatitis delta virus (HDV) in infected human hepatocytes involves association of the 1,679- nucleotide single-stranded genomic RNA (deltaRNA) with multiple copies of both small and large forms of the delta protein (deltaAg) to form a ribonucleoprotein particle which in turn interacts with envelope proteins of the natural helper virus, hepatitis B virus. Subsequently, for initiation of a new round of replication, the amount of small deltaAg within the assembled HDV particle is both necessary and sufficient. Quantitative assays were used in order to better understand just how much deltaAg is needed. The molar ratio of deltaAg species to genomic deltaRNA in assembled HDV particles was approximately 200. Next, this ratio was determined for cells under several different experimental situations in which HDV genome replication was occurring. These included replication in woodchuck liver and also in mouse liver and skeletal muscle, as well as replication in stably and transiently transfected cultured human hepatoblastoma cells. Surprisingly, in almost all these situations the molar ratios were comparable to that observed for HDV particles. This was true for different times after the initiation of replication and was independent of whether or not virus assembly was occurring. Cell fractionation combined with quantitative assays was used to test whether the majority of deltaAg and deltaRNA were colocalized during HDV replication in transfected cells. The cytoplasmic fraction contained the majority of deltaAg and genomic deltaRNA. Finally, the quality of deltaAg and deltaRNA, especially at relatively late times after the initiation of replication, was examined by using reverse transcription-PCR, cloning, and sequencing through the entire deltaAg open reading frame. When virus assembly and spread were not possible, 20% or less of the predicted deltaAg would have been able to support HDV replication. In summary, an examination of the quantity, quality and intracellular distribution of deltaAg and deltaRNA in several different experimental systems has provided a better understanding of the parameters associated with the initiation, maintenance, and ultimate decline of HDV genome replication. PMID- 11907211 TI - Subcellular localization and topology of the p7 polypeptide of hepatitis C virus. AB - Although biological and biochemical data have been accumulated on most hepatitis C virus proteins, the structure and function of the 63-amino-acid p7 polypeptide of this virus have never been investigated. In this work, sequence analyses predicted that p7 contains two transmembrane passages connected by a short hydrophilic segment. The C-terminal transmembrane domain of p7 was predicted to function as a signal sequence, which was confirmed experimentally by analyzing the translocation of a reporter glycoprotein fused at its C terminus. The p7 polypeptide was tagged either with the ectodomain of CD4 or with a Myc epitope to study its membrane integration, its subcellular localization, and its topology. Alkaline extraction studies confirmed that p7 is an integral membrane polypeptide. The CD4-p7 chimera was detected by immunofluorescence on the surface of nonpermeabilized cells, indicating that it is exported to the plasma membrane. However, pulse-chase analyses showed that only approximately 20% of endoglycosidase H-resistant CD4-p7 was detected after long chase times, suggesting that a large proportion of p7 stays in an early compartment of the secretory pathway. Finally, by inserting a Myc epitope in several positions of p7 and analyzing the accessibility of this epitope on the plasma membrane of HepG2 cells, we showed that p7 has a double membrane-spanning topology, with both its N and C termini oriented toward the extracellular environment. Altogether, these data indicate that p7 is a polytopic membrane protein that could have a functional role in several compartments of the secretory pathway. PMID- 11907213 TI - Human immunodeficiency virus cDNA metabolism: notable stability of two-long terminal repeat circles. AB - Early steps of retroviral replication involve reverse transcription of the viral RNA to yield a linear double-stranded cDNA copy and then integration of the viral cDNA into a chromosome of the host cell. A portion of the viral cDNA can also follow nonproductive pathways in which it becomes circularized. In one pathway, the ends of the linear cDNA become joined together by the cellular nonhomologous DNA end-joining system to form two-long terminal repeat (2-LTR) circles. It has been argued that 2-LTR circles are quickly degraded in human immunodeficiency virus (HIV)-infected cells, allowing the presence of 2-LTR circles to be used as a marker for ongoing de novo infection in patients. Following this idea, detection of 2-LTR circles in patients undergoing successful highly active antiretroviral therapy has led to the proposal that viral replication persists despite treatment. We have used fluorescence-monitored PCR (Taqman) to quantitate the metabolism of HIV cDNA early after infection. Contrary to previous work, we find that 2-LTR circles are actually quite stable in experiments where confounding variables are controlled. Thus, studies relying on the lability of 2 LTR circles are open to reinterpretation. We also used the quantitative PCR methods to analyze the effects of MG132, a proteasome inhibitor, which revealed that viral complexes containing mostly completed cDNAs are the primary substrates for proteasome-mediated degradation. PMID- 11907212 TI - Human cytomegalovirus major immediate-early proteins and simian virus 40 large T antigen can inhibit apoptosis through activation of the phosphatidylinositide 3' OH kinase pathway and the cellular kinase Akt. AB - The temperature-sensitive cell line ts13 is mutated in CCG1, the gene encoding TAF(II)250, the largest of the TATA-binding protein-associated factors (TAFs) in TFIID. At the nonpermissive temperature, the temperature-sensitive phenotypes are (i) transcription defects, (ii) cell cycle arrest in G(1), and (iii) apoptosis. We previously demonstrated that the human cytomegalovirus (HCMV) major immediate early proteins (MIEPs) can rescue the transcription defects and inhibit apoptosis at the nonpermissive temperature. In the work presented, we show that activation of the cellular kinase Akt alone can inhibit apoptosis in ts13 cells grown at the nonpermissive temperature. More significantly, we show that the HCMV MIEPs can activate Akt, resulting in the inhibition of apoptosis. In parallel experiments, we found that simian virus 40 (SV40) large T antigen can mediate the same function. These experiments were done by transfecting the HCMV major immediate early gene or a cDNA encoding T antigen into ts13 cells, and thus neither viral attachment to receptors, viral tegument proteins, nor any other viral protein is required for Akt activation. Akt is activated by the phosphatidylinositide 3'-OH (PI3) kinase pathway. Using a specific inhibitor of PI3 kinase, we show that the ability of the MIEPs and T antigen to activate Akt and inhibit apoptosis is eliminated, suggesting that the viral proteins utilize the PI3 kinase pathway for Akt activation. Transfection of plasmids which express the individual 86-kDa (IEP86; IE2(579aa)) and 72-kDa (IEP72; IE1(491aa)) MIEPs indicate that each MIEP could inhibit apoptosis via activation of the PI3 kinase pathway. PMID- 11907214 TI - Sustained high frequencies of specific CD4 T cells restricted to a single persistent virus. AB - Replication of cytomegalovirus (CMV) is largely controlled by the cellular arm of the immune response. In this study the CMV-specific CD4 T-cell response was characterized in a cohort of apparently healthy individuals. In 11% of all individuals, extremely high frequencies, between 10 and 40%, were found. High level frequencies of CMV-specific CD4 T cells persisted over several months and were not the result of an acute infection. Specific T cells were oligoclonal and were phenotypically and functionally characterized as mature effector cells, with both cytokine-secreting and proliferative potential. These high-level frequencies do not seem to compromise the immune response towards heterologous infections, and no signs of immunopathology were observed. Whereas a large temporary expansion of virus-specific T cells is well known to occur during acute infection, we now show that extremely high frequencies of virus-specific T cells may continuously exist in chronic CMV infection without overtly compromising the remaining protective immunity. PMID- 11907215 TI - Membrane recognition by vesicular stomatitis virus involves enthalpy-driven protein-lipid interactions. AB - Vesicular stomatitis virus (VSV) infection depends on the fusion of viral and cellular membranes, which is mediated by virus spike glycoprotein G at the acidic environment of the endosomal compartment. VSV G protein does not contain a hydrophobic amino acid sequence similar to the fusion peptides found among other viral glycoproteins, suggesting that membrane recognition occurs through an alternative mechanism. Here we studied the interaction between VSV G protein and liposomes of different phospholipid composition by force spectroscopy, isothermal titration calorimetry (ITC), and fluorescence spectroscopy. Force spectroscopy experiments revealed the requirement for negatively charged phospholipids for VSV binding to membranes, suggesting that this interaction is electrostatic in nature. In addition, ITC experiments showed that VSV binding to liposomes is an enthalpically driven process. Fluorescence data also showed the lack of VSV interaction with the vesicles as well as inhibition of VSV-induced membrane fusion at high ionic strength. Intrinsic fluorescence measurements showed that the extent of G protein conformational changes depends on the presence of phosphatidylserine (PS) on the target membrane. Although the increase in PS content did not change the binding profile, the rate of the fusion reaction was remarkably increased when the PS content was increased from 25 to 75%. On the basis of these data, we suggest that G protein binding to the target membrane essentially depends on electrostatic interactions, probably between positive charges on the protein surface and negatively charged phospholipids in the cellular membrane. In addition, the fusion is exothermic, indicating no entropic constraints to this process. PMID- 11907216 TI - Analysis of hantavirus genetic diversity in Argentina: S segment-derived phylogeny. AB - Nucleotide sequences were determined for the complete S genome segments of the six distinct hantavirus genotypes from Argentina and for two cell culture isolated Andes virus strains from Chile. Phylogenetic analysis indicates that, although divergent from each other, all Argentinian hantavirus genotypes group together and form a novel phylogenetic clade with the Andes virus. The previously characterized South American hantaviruses Laguna Negra virus and Rio Mamore virus make up another clade that originates from the same ancestral node as the Argentinian/Chilean viruses. Within the clade of Argentinian/Chilean viruses, three subclades can be defined, although the branching order is somewhat obscure. These are made of (i) "Lechiguanas-like" virus genotypes, (ii) Maciel virus and Pergamino virus genotypes, and (iii) strains of the Andes virus. Two hantavirus genotypes from Brazil, Araraquara and Castello dos Sonhos, were found to group with Maciel virus and Andes virus, respectively. The nucleocapsid protein amino acid sequence variability among the members of the Argentinian/Chilean clade does not exceed 5.8%. It is especially low (3.5%) among oryzomyine species-associated virus genotypes, suggesting recent divergence from the common ancestor. Interestingly, the Maciel and Pergamino viruses fit well with the rest of the clade although their hosts are akodontine rodents. Taken together, these data suggest that under conditions in which potential hosts display a high level of genetic diversity and are sympatric, host switching may play a prominent role in establishing hantavirus genetic diversity. However, cospeciation still remains the dominant factor in the evolution of hantaviruses. PMID- 11907217 TI - Improved primate foamy virus vectors and packaging constructs. AB - Foamy virus (FV) vectors that have minimal cis-acting sequences and are devoid of residual viral gene expression were constructed and analyzed by using a packaging system based on transient cotransfection of vector and different packaging plasmids. Previous studies indicated (i) that FV gag gene expression requires the presence of the R region of the long terminal repeat and (ii) that RNA from packaging constructs is efficiently incorporated into vector particles. Mutants with changes in major 5' splice donor (SD) site located in the R region identified this sequence element as responsible for regulating gag gene expression by an unidentified mechanism. Replacement of the FV 5' SD with heterologous splice sites enabled expression of the gag and pol genes. The incorporation of nonvector RNA into vector particles could be reduced to barely detectable levels with constructs in which the human immunodeficiency virus 5' SD or an unrelated intron sequence was substituted for the FV 5' untranslated region and in which gag expression and pol expression were separated on two different plasmids. By this strategy, efficient vector transfer was achieved with constructs that have minimal genetic overlap. PMID- 11907218 TI - Membrane interactions of the tick-borne encephalitis virus fusion protein E at low pH. AB - Membrane fusion of the flavivirus tick-borne encephalitis virus is triggered by the mildly acidic pH of the endosome and is mediated by envelope protein E, a class II viral fusion protein. The low-pH trigger induces an oligomeric rearrangement in which the subunits of the native E homodimers dissociate and the monomeric subunits then reassociate into homotrimers. Here we provide evidence that membrane binding is mediated by the intermediate monomeric form of E, generated by low-pH-induced dissociation of the dimer. Liposome coflotation experiments revealed that association with target membranes occurred only when liposomes were present at the time of acidification, whereas pretreating virions at low pH in the absence of membranes resulted in the loss of their ability to stably attach to liposomes. With the cleavable cross-linker ethylene glycolbis(succinimidylsuccinate), it was shown that a truncated soluble form of the E protein (sE) could bind to membranes only when the dimers were free to dissociate at low pH, and binding could be blocked by a monoclonal antibody that recognizes the fusion peptide, which is at the distal tip of the E monomer but is buried in the native dimer. Surprisingly, analysis of the membrane-associated sE proteins revealed that they had formed trimers. This was unexpected because this protein lacks a sequence element in the C-terminal stem-anchor region, which was shown to be essential for trimerization in the absence of a target membrane. It can therefore be concluded that the formation of a trimeric form of sE is facilitated by membrane binding. Its stability is apparently maintained by contacts between the ectodomains only and is not dependent on sequence elements in the stem-anchor region as previously assumed. PMID- 11907219 TI - Kunjin virus replicon vaccine vectors induce protective CD8+ T-cell immunity. AB - The ability of self-replicating RNA (replicon) vaccine vectors derived from the Australian flavivirus Kunjin (KUN) to induce protective alphabeta CD8+ T-cell responses was examined. KUN replicons encoding a model immunogen were delivered by three different vaccine modalities: (i) as naked RNA transcribed in vitro, (ii) as plasmid DNA constructed to allow in vivo transcription of replicon RNA by cellular RNA polymerase II (DNA based), and (iii) as replicon RNA encapsidated into virus-like particles. A single immunization with any of these KUN replicon vaccines induced CD8+ T-cell responses at levels comparable to those induced by recombinant vaccinia virus encoding the same immunogen. Immunization with only 0.1 microg of DNA-based KUN replicons elicited CD8+ T-cell responses similar to those seen after immunization with 100 microg of a conventional DNA vaccine. Naked RNA immunization with KUN replicons also protected mice against challenges with recombinant vaccinia virus and B16 tumor cells. These results demonstrate the value of KUN replicon vectors for inducing protective antiviral and anticancer CD8+ T-cell responses. PMID- 11907220 TI - Outcome of simian-human immunodeficiency virus strain 89.6p challenge following vaccination of rhesus macaques with human immunodeficiency virus Tat protein. AB - The regulatory proteins Nef, Rev, and Tat of human immunodeficiency virus type 1 (HIV-1) are attractive targets for vaccine development, since induction of effective immune responses targeting these early proteins may best control virus replication. Here we investigated whether vaccination with biologically active Tat or inactive Tat toxoid derived from HIV-1(IIIB) and simian-human immunodeficiency virus (SHIV) strain 89.6p would induce protective immunity in rhesus macaques. Vaccination induced high titers of anti-Tat immunoglobulin G in all immunized animals by week 7, but titers were somewhat lower in the 89.6p Tat group. Dominant B-cell epitopes mapped to the amino terminus, the basic domain, and the carboxy-terminal region. Tat-specific T-helper responses were detected in 50% of immunized animals. T-cell epitopes appeared to map within amino acids (aa) 1 to 24 and aa 37 to 66. In addition, Tat-specific gamma interferon responses were detected in CD4+ and/or CD8+ T lymphocytes in 11 of 16 immunized animals on the day of challenge. However, all animals became infected upon intravenous challenge with 30 50% minimal infective doses of SHIV 89.6p, and there were no significant differences in viral loads or CD4+ T-cell counts between immunized and control animals. Thus, vaccination with HIV-1(IIIB) or SHIV 89.6p Tat or with Tat toxoid preparations failed to confer protection against SHIV 89.6p infection despite robust Tat-specific humoral and cellular immune responses in some animals. Given its apparent immunogenicity, Tat may be more effective as a component of a cocktail vaccine in combination with other regulatory and/or structural proteins of HIV-1. PMID- 11907221 TI - Effects of promyelocytic leukemia protein on virus-host balance. AB - The cellular promyelocytic leukemia protein (PML) associates with the proteins of several viruses and in some cases reduces viral propagation in cell culture. To examine the role of PML in vivo, we compared immune responses and virus loads of PML-deficient and control mice infected with lymphocytic choriomeningitis virus (LCMV) and vesicular stomatitis virus (VSV). PML(-/-) mice exhibited accelerated primary footpad swelling reactions to very-low-dose LCMV, higher swelling peaks upon high-dose inoculation, and higher viral loads in the early phase of systemic LCMV infection. T-cell-mediated hepatitis and consequent mortality upon infection with a hepatotropic LCMV strain required 10- to 100-times-lower inocula despite normal cytotoxic T-lymphocyte reactivity in PML(-/-) mice. Furthermore, PML deficiency rendered mice 10 times more susceptible to lethal immunopathology upon intracerebral LCMV inoculation. Accordingly, 10-times-lower VSV inocula elicited specific neutralizing-antibody responses, a replication-based effect not observed with inactivated virus or after immunization with recombinant VSV glycoprotein. These in vivo observations corroborated our results showing more virus production in PML(-/-) fibroblasts. Thus, PML is a contributor to innate immunity, defining host susceptibility to viral infections and to immunopathology. PMID- 11907222 TI - Increased RNA editing and inhibition of hepatitis delta virus replication by high level expression of ADAR1 and ADAR2. AB - Hepatitis delta virus (HDV) is a subviral human pathogen that uses specific RNA editing activity of the host to produce two essential forms of the sole viral protein, hepatitis delta antigen (HDAg). Editing at the amber/W site of HDV antigenomic RNA leads to the production of the longer form (HDAg-L), which is required for RNA packaging but which is a potent trans-dominant inhibitor of HDV RNA replication. Editing in infected cells is thought to be catalyzed by one or more of the cellular enzymes known as adenosine deaminases that act on RNA (ADARs). We examined the effects of increased ADAR1 and ADAR2 expression on HDV RNA editing and replication in transfected Huh7 cells. We found that both ADARs dramatically increased RNA editing, which was correlated with strong inhibition of HDV RNA replication. While increased HDAg-L production was the primary mechanism of inhibition, we observed at least two additional means by which ADARs can suppress HDV replication. High-level expression of both ADAR1 and ADAR2 led to extensive hyperediting at non-amber/W sites and subsequent production of HDAg variants that acted as trans-dominant inhibitors of HDV RNA replication. Moreover, we also observed weak inhibition of HDV RNA replication by mutated forms of ADARs defective for deaminase activity. Our results indicate that HDV requires highly regulated and selective editing and that the level of ADAR expression can play an important role: overexpression of ADARs inhibits HDV RNA replication and compromises virus viability. PMID- 11907223 TI - Functional interaction between JC virus late regulatory agnoprotein and cellular Y-box binding transcription factor, YB-1. AB - Human polyomavirus JC virus (JCV) is a causative agent of progressive multifocal leukoencephalopathy which results from lytic infection of glial cells. Although significant progress has been made in understanding the regulation of JCV gene transcription, the mechanism(s) underlying the viral lytic cycle remains largely unknown. We recently reported that the JCV late auxiliary Agnoprotein may have a regulatory role in JCV gene transcription and replication. Here, we investigated its regulatory function in viral gene transcription through its physical and functional interaction with YB-1, a cellular transcription factor which contributes to JCV gene expression in glial cells. Time course studies revealed that Agnoprotein is first detected at day 3 postinfection and that its level increased during the late stage of the infection cycle. Agnoprotein is mainly localized to the cytoplasmic compartment of the infected cell, with high concentrations found in the perinuclear region. While the position of Agnoprotein throughout the infection cycle remained relatively unaltered, the subcellular distribution of YB-1 between the cytoplasm and nucleus changed. Results from coimmunoprecipitation and glutathione S-transferase pull-down experiments revealed that Agnoprotein physically interacts with YB-1 and that the amino terminal region of Agnoprotein, between residues 1 and 36, is critical for this association. Further investigation of this interaction by functional assays demonstrated that Agnoprotein negatively regulates YB-1-mediated gene transcription and that the region corresponding to residues 1 to 36 of Agnoprotein is important for the observed regulatory event. Taken together, these data demonstrate that the interaction of the viral late regulatory Agnoprotein and cellular Y-box binding factor YB-1 modulates transcriptional activity of JCV promoters. PMID- 11907224 TI - Bovine herpesvirus 5 (BHV-5) Us9 is essential for BHV-5 neuropathogenesis. AB - Bovine herpesvirus 5 (BHV-5) is a neurovirulent alphaherpesvirus that causes fatal encephalitis in calves. In a rabbit model, the virus invades the central nervous system (CNS) anterogradely from the olfactory mucosa following intranasal infection. In addition to glycoproteins E and I (gE and gI, respectively), Us9 and its homologue in alphaherpesviruses are necessary for the viral anterograde spread from the presynaptic to postsynaptic neurons. The BHV-5 Us9 gene sequence was determined, and the predicted amino acid sequence of BHV-5 Us9 was compared with the corresponding Us9 sequences of BHV-1.1. Alignment results showed that they share 77% identity and 83% similarity. BHV-5 Us9 peptide-specific antibody recognized a doublet of 17- and 19-kDa protein bands in BHV-5-infected cell lysates and in purified virions. To determine the role of the BHV-5 Us9 gene in BHV-5 neuropathogenesis, a BHV-5 Us9 deletion recombinant was generated and its neurovirulence and neuroinvasive properties were compared with those of a Us9 rescue mutant of BHV-5 in a rabbit model. Following intranasal infection, the Us9 rescue mutant of BHV-5 displayed a wild-type level of neurovirulence and neural spread in the olfactory pathway, but the Us9 deletion mutant of BHV-5 was virtually avirulent and failed to invade the CNS. In the olfactory mucosa containing the olfactory receptor neurons, the Us9 deletion mutant virus replicated with an efficiency similar to that of the Us9 rescue mutant of BHV-5. However, the Us9 deletion mutant virus was not transported to the bulb. Confocal microscopy of the olfactory epithelium detected similar amounts of virus-specific antigens in the cell bodies of olfactory receptor neuron for both the viruses, but only the Us9 rescue mutant viral proteins were detected in the processes of the olfactory receptor neurons. When injected directly into the bulb, both viruses were equally neurovirulent, and they were transported retrogradely to areas connected to the bulb. Taken together, these results indicate that Us9 is essential for the anterograde spread of the virus from the olfactory mucosa to the bulb. PMID- 11907226 TI - Oligomerization and cooperative RNA synthesis activity of hepatitis C virus RNA dependent RNA polymerase. AB - The NS5B RNA-dependent RNA polymerase encoded by hepatitis C virus (HCV) plays a key role in viral replication. Reported here is evidence that HCV NS5B polymerase acts as a functional oligomer. Oligomerization of HCV NS5B protein was demonstrated by gel filtration, chemical cross-linking, temperature sensitivity, and yeast cell two-hybrid analysis. Mutagenesis studies showed that the C terminal hydrophobic region of the protein was not essential for its oligomerization. Importantly, HCV NS5B polymerase exhibited cooperative RNA synthesis activity with a dissociation constant, K(d), of approximately 22 nM, suggesting a role for the polymerase-polymerase interaction in the regulation of HCV replicase activity. Further functional evidence includes the inhibition of the wild-type NS5B polymerase activity by a catalytically inactive form of NS5B. Finally, the X-ray crystal structure of HCV NS5B polymerase was solved at 2.9 A. Two extensive interfaces have been identified from the packing of the NS5B molecules in the crystal lattice, suggesting a higher-order structure that is consistent with the biochemical data. PMID- 11907225 TI - Variability in the human immunodeficiency virus type 1 gp120 Env protein linked to phenotype-associated changes in the V3 loop. AB - Isolates of human immunodeficiency virus type 1 (HIV-1) are classified according to the chemokine receptor (coreceptor) used in conjunction with CD4 to target and enter cells: viruses using CCR5 and CXCR4 are classified as R5 and X4, respectively. The major determinant of entry-related HIV-1 phenotypes is known to reside in the third variable region of gp120 (V3). It is clear, however, that positions outside of V3 play some role in influencing phenotype, although marked context dependence and extensive variability among HIV-1 isolates have made the identification of these positions difficult. We used the presence of previously described substitutions in V3 to classify a large set of HIV-1 subtype B gp120 sequences available in public databases as X4-like or R5-like. Using these classifications, we searched for positions outside of V3 where either amino acid composition or variability differed significantly among sequences of different inferred phenotypes. Our approach took the epidemiological relationships among sequences into account. A cluster of positions linked to changes in V3 was identified between amino acids 190 and 204 of gp120, immediately C-terminal of V2; changes at position 440 in C4 were also linked to inferred phenotype. Structural data place these positions at the coreceptor-binding face of gp120 in a surface-exposed location. We also noted a significant increase in net positive charge in a highly variable region of V2. This study both confirms previous observations and predicts specific positions that contribute to a functional relationship between V3, V2, and C4. PMID- 11907227 TI - Varied persistent life cycles of Borna disease virus in a human oligodendroglioma cell line. AB - Borna disease virus (BDV) establishes a persistent infection in the central nervous system of vertebrate animal species as well as in tissue cultures. In an attempt to characterize the life cycle of BDV in persistently infected cultured cells, we developed 30 clones by single-cell cloning from a human oligodendroglioma (OL) cell line after infection with BDV. According to the percentage of cells expressing the BDV major proteins, p40 (nucleoprotein) and p24 (phosphoprotein), the clones were classified into two types: type I (>20%) and type II (<20%). mRNAs corresponding to both proteins were detected by in situ hybridization (ISH) in a percentage of cells consistent with that for the protein expression in the two types. Surprisingly, ISH for the detection of the genomic RNA, mainly in type II, revealed a significantly larger cell population harboring the genomic RNA than that with the protein as well as the mRNA expression. By recloning from type II primary cell clones, the same phenotype was confirmed in the secondary cell clones obtained: i.e., low percentage of protein-positive cells and higher percentage of cells harboring the genomic RNA. After nerve growth factor treatment, the two types of clones showed increases in the percentage of cells expressing BDV-specific proteins that reached 80% in type II clones, in addition to increased expression levels per cell. Such enhancement might have been mediated by the activation of the mitogen-activated protein kinase in the clones as revealed by the detection of activated ERK1/2. Thus, our findings show that BDV may have established a persistent infection at low levels of viral expression in OL cells with the possibility of a latent infection. PMID- 11907228 TI - Anti-TAR polyamide nucleotide analog conjugated with a membrane-permeating peptide inhibits human immunodeficiency virus type 1 production. AB - The emergence of drug-resistant variants has posed a significant setback against effective antiviral treatment for human immunodeficiency virus (HIV) infections. The choice of a nonmutable region of the viral genome such as the conserved transactivation response element (TAR element) in the 5' long terminal repeat (LTR) may potentially be an effective target for drug development. We have earlier demonstrated that a polyamide nucleotide analog (PNA) targeted to the TAR hairpin element, when transfected into cells, can effectively inhibit Tat mediated transactivation of HIV type 1 (HIV-1) LTR (T. Mayhood et al., Biochemistry 39:11532-11539, 2000). Here we show that this anti-TAR PNA (PNA(TAR)), upon conjugation with a membrane-permeating peptide vector (transportan) retained its affinity for TAR in vitro similar to the unconjugated analog. The conjugate was efficiently internalized into the cells when added to the culture medium. Examination of the functional efficacy of the PNA(TAR) transportan conjugate in cell culture using luciferase reporter gene constructs resulted in a significant inhibition of Tat-mediated transactivation of HIV-1 LTR. Furthermore, PNA(TAR)-transportan conjugate substantially inhibited HIV-1 production in chronically HIV-1-infected H9 cells. The mechanism of this inhibition appeared to be regulated at the level of transcription. These results demonstrate the efficacy of PNA(TAR)-transportan as a potential anti-HIV agent. PMID- 11907229 TI - Minute virus of mice NS1 interacts with the SMN protein, and they colocalize in novel nuclear bodies induced by parvovirus infection. AB - The human survival motor neuron (SMN) gene is the spinal muscular atrophy determining gene, and a knockout of the murine Smn gene results in preembryonic lethality. Here we show that SMN can directly interact in vitro and in vivo with the large nonstructural protein NS1 of the autonomous parvovirus minute virus of mice (MVM), a protein essential for viral replication and a potent transcriptional activator. Typically, SMN localizes within nuclear Cajal bodies and diffusely in the cytoplasm. Following transient NS1expression, SMN and NS1 colocalize within Cajal bodies. At early time points following parvovirus infection, NS1 fails to colocalize with SMN within Cajal bodies; however, during the course of MVM infection, dramatic nuclear alterations occur. Formerly distinct nuclear bodies such as Cajal bodies, promyelocytic leukemia gene product (PML) oncogenic domains (PODs), speckles, and autonomous parvovirus-associated replication (APAR) bodies are seen aggregating at later points in infection. These newly formed large nuclear bodies (termed SMN-associated APAR bodies) are active sites of viral replication and viral capsid assembly. These results highlight the transient nature of nuclear bodies and their contents and identify a novel nuclear body formed during infection. Furthermore, simple transient expression of the viral nonstructural proteins is insufficient to induce this nuclear reorganization, suggesting that this event is induced specifically by a step in the viral infection process. PMID- 11907230 TI - Long-distance base pairing in flock house virus RNA1 regulates subgenomic RNA3 synthesis and RNA2 replication. AB - Replication of flock house virus (FHV) RNA1 and production of subgenomic RNA3 in the yeast Saccharomyces cerevisiae provide a useful tool for the dissection of FHV molecular biology and host-encoded functions involved in RNA replication. The replication template activity of RNA1 can be separated from its coding potential by supplying the RNA1-encoded replication factor protein A in trans. We constructed a trans-replication system in yeast to examine cis-acting elements in RNA1 that control RNA3 production, as well as RNA1 and RNA2 replication. Two cis elements controlling RNA3 production were found. A proximal subgenomic control element was located just upstream of the RNA3 start site (nucleotides [nt] 2282 to 2777). A short distal element also controlling RNA3 production (distal subgenomic control element) was identified 1.5 kb upstream, at nt 1229 to 1239. Base pairing between these distal and proximal elements was shown to be essential for RNA3 production by covariation analysis and in vivo selection of RNA3 expressing replicons from plasmid libraries containing random sequences in the distal element. Two distinct RNA1 replication elements (RE) were mapped within the 3' quarter of RNA1: the intRE (nt 2322 to 2501) and the 3'RE (nt 2735 to 3011). The 3'RE significantly overlaps the RNA3 region in RNA1, and this information was applied to produce improved RNA3-based vectors for foreign-gene expression. In addition, replication of an RNA2 derivative was dependent on RNA1 templates capable of forming the long-distance interaction that controls RNA3 production. PMID- 11907231 TI - Rolling circle replication of hepatitis delta virus RNA is carried out by two different cellular RNA polymerases. AB - Hepatitis delta virus (HDV) contains a viroid-like circular RNA that is presumed to replicate via a rolling circle replication mechanism mediated by cellular RNA polymerases. However, the exact mechanism of rolling circle replication for HDV RNA and viroids is not clear. Using our recently described cDNA-free transfection system (L. E. Modahl and M. M. Lai, J. Virol. 72:5449-5456, 1998), we have succeeded in detecting HDV RNA replication by metabolic labeling with [32P]orthophosphate in vivo and obtained direct evidence that HDV RNA replication generates high-molecular-weight multimeric species of HDV RNA, which are processed into monomeric and dimeric forms. Thus, these multimeric RNAs are the true intermediates of HDV RNA replication. We also found that HDV RNA synthesis is highly temperature sensitive, occurring most efficiently at 37 to 40 degrees C and becoming virtually undetectable at temperatures below 30 degrees C. Moreover, genomic HDV RNA synthesis was found to occur at a rate roughly 30-fold higher than that of antigenomic RNA synthesis. Finally, in lysolecithin-permeabilized cells, the synthesis of full-length antigenomic HDV RNA was completely resistant to high concentrations (100 microg/ml) of alpha-amanitin. In contrast, synthesis of genomic HDV RNA was totally inhibited by alpha-amanitin at concentrations as low as 2.5 microg/ml. Thus, these results suggest that genomic and antigenomic HDV RNA syntheses are performed by two different host cell enzymes. This observation, combined with our previous finding that hepatitis delta antigen mRNA synthesis is likely performed by RNA polymerase II, suggests that the different HDV RNA species are synthesized by different cellular transcriptional machineries. PMID- 11907232 TI - Genomic but not antigenomic hepatitis delta virus RNA is preferentially exported from the nucleus immediately after synthesis and processing. AB - Hepatitis delta virus (HDV) contains a viroid-like circular RNA that replicates via a double rolling circle replication mechanism. It is generally assumed that HDV RNA is synthesized and remains exclusively in the nucleus until being exported to the cytoplasm for virion assembly. Using a [32P]orthophosphate metabolic labeling procedure to study HDV RNA replication (T. B. Macnaughton, S. T. Shi, L. E. Modahl, and M. M. C. Lai. J. Virol. 76:3920-3927, 2002), we unexpectedly found that a significant amount of newly synthesized HDV RNA was detected in the cytoplasm. Surprisingly, Northern blot analysis revealed that the genomic-sense HDV RNA is present almost equally in both the nucleus and cytoplasm, whereas antigenomic HDV RNA was mostly retained in the nucleus, suggesting the specific and highly selective export of genomic HDV RNA. Kinetic studies showed that genomic HDV RNA was exported soon after synthesis. However, only the monomer and, to a lesser extent, the dimer HDV RNAs were exported to the cytoplasm; very little higher-molecular-weight HDV RNA species were detected in the cytoplasm. These results suggest that the cleavage and processing of HDV RNA may facilitate RNA export. The export of genomic HDV RNA was resistant to leptomycin B, indicating that a cell region maintenance 1 (Crm1)-independent pathway was involved. The large form of hepatitis delta antigen (L-HDAg), which is responsible for virus packaging, was not required for RNA export, as a mutant HDV RNA genome unable to synthesize L-HDAg was still exported. The proportions of genomic HDV RNA in the nucleus and cytoplasm remained relatively constant throughout replication, indicating that export of genomic HDV RNA occurred continuously. In contrast, while antigenomic HDV RNA was predominantly in the nucleus, there was a proportionally large fraction of antigenomic HDV RNA in the cytoplasm at early time points of RNA replication. These findings uncover a previously unrecognized presence of HDV RNA in the cytoplasm, which may have implications for viral RNA synthesis and packaging. PMID- 11907233 TI - African swine fever virus IAP-like protein induces the activation of nuclear factor kappa B. AB - African swine fever virus (ASFV) encodes a homologue of the inhibitor of apoptosis (IAP) that promotes cell survival by controlling the activity of caspase-3. Here we show that ASFV IAP is also able to activate the transcription factor NF-kappaB. Thus, transient transfection of the viral IAP increases the activity of an NF-kappaB reporter gene in a dose-responsive manner in Jurkat cells. Similarly, stably transfected cells expressing ASFV IAP have elevated basal levels of c-rel, an NF-kappaB-dependent gene. NF-kappaB complexes in the nucleus were increased in A224L-expressing cells compared with control cells upon stimulation with phorbol myristate acetate (PMA) plus ionomycin. This resulted in greater NF-kappaB-dependent promoter activity in ASFV IAP-expressing than in control cells, both in basal conditions and after PMA plus ionophore stimulation. The elevated NF-kappaB activity seems to be the consequence of higher IkappaB kinase (IKK) basal activity in these cells. The NF-kappaB-inducing activity of ASFV IAP was abrogated by an IKK-2 dominant negative mutant and enhanced by expression of tumor necrosis factor receptor-associated factor 2. PMID- 11907234 TI - Role of lymphotoxin alpha in T-cell responses during an acute viral infection. AB - The importance of lymphotoxin alpha (LTalpha) in lymphoid organogenesis is well established. Although LTalpha has been implicated in the pathogenesis of T-cell mediated immunopathologies, the requirement for LTalpha in T-cell activation and effector function in vivo is not well understood. To determine the role of LTalpha in T-cell activation in vivo, we compared the generation of antigen specific T-cell responses between wild type (+/+) and LTalpha-deficient (LTalpha( /-)) mice during an acute infection with lymphocytic choriomeningitis virus (LCMV). Our studies showed that LCMV-infected LTalpha(-/-) mice had a profound impairment in the activation and expansion of virus-specific CD8 T cells in the spleen, as determined by cytotoxicity assays, intracellular staining for gamma interferon, and staining with major histocompatibility complex class I tetramers. Further, the nonlymphoid organs of LTalpha(-/-) mice also contained substantially lower number of LCMV-specific CD8 T cells than those of +/+ mice. Greatly reduced virus-specific CD8 T-cell responses in LTalpha(-/-) mice led to a defect in LCMV clearance from the tissues. In comparison to that in +/+ mice, the activation of LCMV-specific CD4 T cells was also significantly attenuated in LTalpha(-/-) mice. Adoptive transfer experiments were conducted to determine if abnormal lymphoid architecture in LTalpha(-/-) mice caused the impairment in the activation of LCMV specific T-cell responses. Upon adoptive transfer into +/+ mice, the activation and expansion of LCMV-specific LTalpha(-/-) T cells were restored to levels comparable to those of +/+ T cells. In a reciprocal cell transfer experiment, activation of +/+ T cells was significantly reduced upon transfer into LTalpha(-/ ) mice. These results showed that impairment in the activation of LCMV-specific T cells in LTalpha(-/-) mice may be due to abnormal lymphoid architecture and not to an intrinsic defect in LTalpha(-/-) T cells. PMID- 11907235 TI - Requirements for budding of paramyxovirus simian virus 5 virus-like particles. AB - Enveloped viruses are released from infected cells after coalescence of viral components at cellular membranes and budding of membranes to release particles. For some negative-strand RNA viruses (e.g., vesicular stomatitis virus and Ebola virus), the viral matrix (M) protein contains all of the information needed for budding, since virus-like particles (VLPs) are efficiently released from cells when the M protein is expressed from cDNA. To investigate the requirements for budding of the paramyxovirus simian virus 5 (SV5), its M protein was expressed in mammalian cells, and it was found that SV5 M protein alone could not induce vesicle budding and was not secreted from cells. Coexpression of M protein with the viral hemagglutinin-neuraminidase (HN) or fusion (F) glycoproteins also failed to result in significant VLP release. It was found that M protein in the form of VLPs was only secreted from cells, with an efficiency comparable to authentic virus budding, when M protein was coexpressed with one of the two glycoproteins, HN or F, together with the nucleocapsid (NP) protein. The VLPs appeared similar morphologically to authentic virions by electron microscopy. CsCl density gradient centrifugation indicated that almost all of the NP protein in the cells had assembled into nucleocapsid-like structures. Deletion of the F and HN cytoplasmic tails indicated an important role of these cytoplasmic tails in VLP budding. Furthermore, truncation of the HN cytoplasmic tail was found to be inhibitory toward budding, since it prevented coexpressed wild-type (wt) F protein from directing VLP budding. Conversely, truncation of the F protein cytoplasmic tail was not inhibitory and did not affect the ability of coexpressed wt HN protein to direct the budding of particles. Taken together, these data suggest that multiple viral components, including assembled nucleocapsids, have important roles in the paramyxovirus budding process. PMID- 11907236 TI - High frequency of virus-specific B lymphocytes in germinal centers of simian human immunodeficiency virus-infected rhesus monkeys. AB - The etiology of the lymphadenopathy and follicular hyperplasia associated with human immunodeficiency virus type 1 (HIV-1) infection has remained unclear. To determine whether the B-lymphocyte expansions characteristic of this syndrome represent polyclonal and virus-specific processes, the antigen specificity of B cells in lymphoid tissues of monkeys infected with simian-human immunodeficiency virus (SHIV) chimeras was assessed using an inverse immunohistochemical assay with biotinylated HIV-1 envelope gp120 (Env) as an antigen probe. Env-binding B cells were found aggregated in lymph node and splenic germinal centers (GCs). Most Env-binding GCs also contained an unstained population of B cells, suggesting the GCs were formed by a polyclonal (oligoclonal) process. By day 42 following infection, Env-binding B cells were present in 19% of all lymph node GCs. Env-binding cells were present in 25% of GCs even during chronic infection. This extraordinarily high frequency of Env-specific B lymphocytes suggests that the expansion of virus-specific B cells may largely account for the follicular hyperplasia in AIDS virus-infected individuals. PMID- 11907237 TI - Intramolecular complementing mutations in tobacco mosaic virus movement protein confirm a role for microtubule association in viral RNA transport. AB - The movement protein (MP) of Tobacco mosaic virus (TMV) facilitates the cell-to cell transport of the viral RNA genome through plasmodesmata (Pd). A previous report described the functional reversion of a dysfunctional mutation in MP (Pro81Ser) by two additional amino acid substitution mutations (Thr104Ile and Arg167Lys). To further explore the mechanism underlying this intramolecular complementation event, the mutations were introduced into a virus derivative expressing the MP as a fusion to green fluorescent protein (GFP). Microscopic analysis of infected protoplasts and of infection sites in leaves of MP transgenic Nicotiana benthamiana indicates that MP(P81S)-GFP and MP(P81S;T104I;R167K)-GFP differ in subcellular distribution. MP(P81S)-GFP lacks specific sites of accumulation in protoplasts and, in epidermal cells, exclusively localizes to Pd. MP(P81S;T104I;R167K)-GFP, in contrast, in addition localizes to inclusion bodies and microtubules and thus exhibits a subcellular localization pattern that is similar, if not identical, to the pattern reported for wild-type MP-GFP. Since accumulation of MP to inclusion bodies is not required for function, these observations confirm a role for microtubules in TMV RNA cell-to-cell transport. PMID- 11907238 TI - Expression of simian immunodeficiency virus nef in immune cells of transgenic mice leads to a severe AIDS-like disease. AB - In order to study the functions of simian immunodeficiency virus (SIV) Nef in vivo in a small-animal model, we constructed transgenic (Tg) mice expressing the SIV(mac)239 nef gene in the natural target cells of the virus under the control of the human CD4 gene promoter (CD4C). These CD4C/SHIV-nef(SIV) Tg mice develop a severe AIDS-like disease, with manifestations including premature death, failure to thrive or weight loss, wasting, thymic atrophy, an especially low number of peripheral CD8+ T cells as well as a low number of peripheral CD4+ T cells, diarrhea, splenomegaly, and kidney (interstitial nephritis, segmental glomerulosclerosis), lung (lymphocytic interstitial pneumonitis), and heart disease. In addition, these Tg mice fail to mount a class-switched antibody response after immunization with ovalbumin, they produce anti-DNA autoantibodies, and some of them develop Pneumocystis carinii lung infections. All these results suggest a generalized Nef-induced immunodeficiency. The low numbers of peripheral CD8+ and CD4+ T cells are likely to reflect a thymic defect and may be similar to the DiGeorge-like "thymic defect" immunophenotype described for a subgroup of human immunodeficiency virus type 1-infected children. Therefore, it appears that SIV Nef alone expressed in mice, in appropriate cell types and at sufficient levels, can elicit many of the phenotypes of simian and human AIDS. These Tg mice should be instrumental in studying the pathogenesis of SIV Nef-induced phenotypes. PMID- 11907239 TI - Transcriptional targeting of lentiviral vectors by long terminal repeat enhancer replacement. AB - Gene therapy of many genetic diseases requires permanent gene transfer into self renewing stem cells and restriction of transgene expression to specific progenies. Human immunodeficiency virus (HIV)-derived lentiviral vectors are very effective in transducing rare, nondividing stem cell populations (e.g., hematopoietic stem cells) without altering their long-term repopulation and differentiation capacities. We developed a strategy for transcriptional targeting of lentiviral vectors based on replacing the viral long terminal repeat (LTR) enhancer with cell lineage-specific, genomic control elements. An upstream enhancer (HS2) of the erythroid-specific GATA-1 gene was used to replace most of the U3 region of the LTR, immediately upstream of the HIV type 1 (HIV-1) promoter. The modified LTR was used to drive the expression of a reporter gene (the green fluorescent protein [GFP] gene), while a second gene (a truncated form of the p75 nerve growth factor receptor [DeltaLNGFR]) was placed under the control of an internal constitutive promoter to monitor cell transduction, or to immunoselect transduced cells, independently from the expression of the targeted promoter. The transcriptionally targeted vectors were used to transduce cell lines, human CD34+ hematopoietic stem-progenitor cells, and murine bone marrow (BM)-repopulating stem cells. Gene expression was analyzed in the stem cell progeny in vitro and in vivo after xenotransplantation into nonobese diabetic SCID mice or BM transplantation in coisogenic mice. The modified LTR directed high levels of transgene expression specifically in mature erythroblasts, in a TAT-independent fashion and with no alteration in titer, infectivity, and genomic stability of the lentiviral vector. Expression from the modified LTR was higher, better restricted, and showed less position-effect variegation than that obtained by the same combination of enhancer-promoter elements placed in a conventional, internal position. Cloning of the woodchuck hepatitis virus posttranscriptional regulatory element at a defined position in the targeted vector allowed selective accumulation of the genomic transcripts with respect to the internal RNA transcript, with no loss of cell-type restriction. A critical advantage of this targeting strategy is the use of a spliced, major viral transcript to express a therapeutic gene and that of an internal, independently regulated promoter to express an additional gene for either cell marking or in vivo selection purposes. PMID- 11907240 TI - Persistent and transient replication of full-length hepatitis C virus genomes in cell culture. AB - The recently developed subgenomic hepatitis C virus (HCV) replicons were limited by the fact that the sequence encoding the structural proteins was missing. Therefore, important information about a possible influence of these proteins on replication and pathogenesis and about the mechanism of virus formation could not be obtained. Taking advantage of three cell culture-adaptive mutations that enhance RNA replication synergistically, we generated selectable full-length HCV genomes that amplify to high levels in the human hepatoma cell line Huh-7 and can be stably propagated for more than 6 months. The structural proteins are efficiently expressed, with the viral glycoproteins E1 and E2 forming heterodimers which are stable under nondenaturing conditions. No disulfide-linked glycoprotein aggregates were observed, suggesting that the envelope proteins fold productively. Electron microscopy studies indicate that cell lines harboring these full-length HCV RNAs contain lipid droplets. The majority of the core protein was found on the surfaces of these structures, whereas the glycoproteins appear to localize to the endoplasmic reticulum and cis-Golgi compartments. In agreement with this distribution, no endoglycosidase H-resistant forms of these proteins were detectable. In a search for the production of viral particles, we noticed that these cells release substantial amounts of nuclease-resistant HCV RNA-containing structures with a buoyant density of 1.04 to 1.1 g/ml in iodixanol gradients. The same observation was made in transient-replication assays using an authentic highly adapted full-length HCV genome that lacks heterologous sequences. However, the fact that comparable amounts of such RNA-containing structures were found in the supernatant of cells carrying subgenomic replicons demonstrates a nonspecific release independent of the presence of the structural proteins. These results suggest that Huh-7 cells lack host cell factors that are important for virus particle assembly and/or release. PMID- 11907241 TI - Human T-lymphotropic virus type 1 oncoprotein tax promotes S-phase entry but blocks mitosis. AB - Human T-lymphotropic virus type 1 (HTLV-1) Tax exerts pleiotropic effects on multiple cellular regulatory processes to bring about NF-kappaB activation, aberrant cell cycle progression, and cell transformation. Here we report that Tax stimulates cellular G(1)/S entry but blocks mitosis. Tax expression in naive cells transduced with a retroviral vector, pBabe-Tax, leads to a significant increase in the number of cells in the S phase, with an accompanying rise in the population of cells with a DNA content of 4N or more. In all cell types tested, including BHK-21, mouse NIH 3T3, and human diploid fibroblast WI-38, Tax causes an uncoupling of DNA synthesis from cell division, resulting in the formation of multinucleated giant cells and cells with decondensed, highly convoluted and lobulated nuclei that are reminiscent of the large lymphocytes with cleaved or cerebriform nuclei seen in HTLV-1-positive individuals. This contrasts with the Tax-transformed cell lines, PX1 (fibroblast) and MT4 (lymphocyte), which produce Tax at high levels, but without the accompanying late-stage cell cycle abnormalities. PX1 and MT4 may have been selected to harbor somatic mutations that allow a bypass of the Tax-induced block in mitosis. PMID- 11907242 TI - A natural intergenotypic recombinant of hepatitis C virus identified in St. Petersburg. AB - Hepatitis C virus (HCV) evolution is thought to proceed by mutations within the six genotypes. Here, we report on a viable spontaneous HCV recombinant and we show that recombination may play a role in the evolution of this virus. Previously, 149 HCV strains from St. Petersburg had been subtyped by limited sequencing within the NS5B region. In the present study, the core regions of 41 of these strains were sequenced to investigate the concordance of HCV genotyping for these two genomic regions. Two phylogenetically related HCV strains were found to belong to different subtypes, 2k and 1b, according to sequence analysis of the 5' untranslated region (5'UTR)-core and the NS5B regions, respectively. By sequencing of the E2-p7-NS2 region, the crossover point was mapped within the NS2 region, probably between positions 3175 and 3176 (according to the numbering system for strain pj6CF). Sequencing of the 5'UTR-core regions of four other HCV strains, phylogenetically related to the above-mentioned two strains (based on analysis within the NS5B region), revealed that these four strains were also recombinants. Since a nonrecombinant 2k strain was found in St. Petersburg, the recombination may have taken place there around a decade ago. Since the frequency of this recombinant is now high enough to allow the detection of the recombinant in a fraction of the city's population, it seems to be actively spreading there. The reported recombinant is tentatively designated RF1-2k/1b, in agreement with the nomenclature used for HIV recombinants. Recombination between HCV genotypes must now be considered in the classification, laboratory diagnosis, and treatment of HCV infection. PMID- 11907243 TI - Structural requirements for the assembly of Norwalk virus-like particles. AB - Norwalk virus (NV) is the prototype strain of a group of human caliciviruses responsible for epidemic outbreaks of acute gastroenteritis. While these viruses do not grow in tissue culture cells or animal models, expression of the capsid protein in insect cells results in the self-assembly of recombinant NV virus-like particles (rNV VLPs) that are morphologically and antigenically similar to native NV. The X-ray structure of the rNV VLPs has revealed that the capsid protein folds into two principal domains: a shell (S) domain and a protruding (P) domain (B. V. V. Prasad, M. E. Hardy, T. Dokland, J. Bella, M. G. Rossmann, and M. K. Estes, Science 286:287-290, 1999). To investigate the structural requirements for the assembly of rNV VLPs, we performed mutational analyses of the capsid protein. We examined the ability of 10 deletion mutants of the capsid protein to assemble into VLPs in insect cell cultures. Deletion of the N-terminal 20 residues, suggested by the X-ray structure to be involved in a switching mechanism during assembly, did not affect the ability of the mutant capsid protein to self assemble into 38-nm VLPs with a T=3 icosahedral symmetry. Further deletions in the N-terminal region affected particle assembly. Deletions in the C-terminal regions of the P domain, involved in the interactions between the P and S domains, did not block the assembly process, but they affected the size and stability of the particles. Mutants carrying three internal deletion mutations in the P domain, involved in maintaining dimeric interactions, produced significantly larger 45-nm particles, albeit in low yields. The complete removal of the protruding domain resulted in the formation of smooth particles with a diameter that is slightly smaller than the 30-nm diameter expected from the rNV structure. These studies indicate that the shell domain of the NV capsid protein contains everything required to initiate the assembly of the capsid, whereas the entire protruding domain contributes to the increased stability of the capsid by adding intermolecular contacts between the dimeric subunits and may control the size of the capsid. PMID- 11907245 TI - Mutations that confer resistance to template-analog inhibitors of human immunodeficiency virus (HIV) type 1 reverse transcriptase lead to severe defects in HIV replication. AB - We isolated two template analog reverse transcriptase (RT) inhibitor-resistant mutants of human immunodeficiency virus (HIV) type 1 RT by using the DNA aptamer, RT1t49. The mutations associated, N255D or N265D, displayed low-level resistance to RT1t49, while high-level resistance could be observed when both mutations were present (Dbl). Molecular clones of HIV that contained the mutations produced replication-defective virions. All three RT mutants displayed severe processivity defects. Thus, while biochemical resistance to the DNA aptamer RT1t49 can be generated in vitro via multiple mutations, the overlap between the aptamer- and template-primer-binding pockets favors mutations that also affect the RT-template primer interaction. Therefore, viruses with such mutations are replication defective. Potent inhibition and a built-in mechanism to render aptamer-resistant viruses replication defective make this an attractive class of inhibitors. PMID- 11907244 TI - A protein encoded by the herpes simplex virus (HSV) type 1 2-kilobase latency associated transcript is phosphorylated, localized to the nucleus, and overcomes the repression of expression from exogenous promoters when inserted into the quiescent HSV genome. AB - Herpes simplex virus (HSV) is characterized by its ability to establish a latent infection in sensory neurons, from which it can periodically reactivate. The mechanisms of latency, however, remain unclear. The HSV genome is quiescent during latency except for the expression of the latency-associated transcripts (LATs). Although the exact function of the LATs remains obscure, current evidence suggests they are multifunctional and are involved in both establishment of latency and reactivation from latency. The LATs contain several open reading frames (ORFs). One or more of the functions of the LATs could therefore be protein mediated. We have previously reported that deregulated expression of the largest of the HSV type 1 (HSV-1) LAT ORFs ( approximately 274 amino acids) greatly enhances virus growth in cell types that are normally relatively nonpermissive for HSV replication and also that it complements mutations to the immediate-early (IE) gene ICP0 (S. K. Thomas, G. Gough, D. S. Latchman, and R. S. Coffin, J. Virol. 73:6618-6625, 1999). Here we show that LAT ORF expression overcomes the repression of expression from exogenous promoters introduced into the HSV-1 genome which normally occurs in the absence of IE gene expression. To further explore LAT ORF function, we have generated an epitope-tagged LAT ORF, LATmycHis, which forms punctate structures in the infected-cell nucleus reminiscent of the structures formed by ICP0. These are associated with the appearance of a phosphorylated form of the protein and are formed adjacent to, or around the edges of, viral replication compartments. These results provide further evidence that the HSV-1 LAT ORF protein is biologically functional and that the tightly regulated expression of this protein may be important in the wild-type latency phenotype in vivo. PMID- 11907246 TI - Hepatitis C virus-like particle morphogenesis. AB - Although much is known about the hepatitis C virus (HCV) genome, first cloned in 1989, little is known about HCV structure and assembly due to the lack of an efficient in vitro culture system for HCV. Using a recombinant Semliki forest virus replicon expressing genes encoding HCV structural proteins, we observed for the first time the assembly of these proteins into HCV-like particles in mammalian cells. This system opens up new possibilities for the investigation of viral morphogenesis and virus-host cell interactions. PMID- 11907247 TI - Preferential localization of the Epstein-Barr virus (EBV) oncoprotein LMP-1 to nuclei in human T cells: implications for its role in the development of EBV genome-positive T-cell lymphomas. AB - The Epstein-Barr virus (EBV)-encoded latent membrane protein-1 (LMP-1) is thought to play a role in the EBV-induced B-cell transformation and immortalization. EBV has also been implicated in certain human T-cell lymphomas; however, the phenotypic effects of the expression of this oncoprotein in T cells are not known. To learn whether LMP-1 also induces phenotypic changes in T cells, we stably expressed it in human cell lines of T and B lineages and 25 LMP-1 expressing T-cell clones and 7 B-cell clones were examined. Our results show for the first time that, in sharp contrast to B cells, LMP-1 preferentially localizes to nuclei in T cells and does not induce the phenotypic changes in these cells that it induces in B cells, does not associate with TRAF proteins, and does not arrest the cell cycle in the G2/M phase. A computer-assisted analysis revealed that LMP-1 lacks the canonical nuclear localization signal. Our results suggest that this oncoprotein may not play the same role in the lymphomagenesis of T cells as it does in B cells. PMID- 11907248 TI - Initial interaction of rotavirus strains with N-acetylneuraminic (sialic) acid residues on the cell surface correlates with VP4 genotype, not species of origin. AB - We examined 41 human and animal rotavirus strains representative of all known P genotypes for their dependency on cellular N-acetylneuraminic (sialic) acid (SA) residues for infectivity. Our results showed that all rotaviruses studied, whether of animal or human origin, belonging to P genotypes [1], [2], [3], and [7] depended on SA residues on the cell surface for efficient infectivity but that all human and animal rotavirus strains representative of the remaining known P genotypes were SA independent. The SA residue requirement for efficient infectivity did not change for reassortant rotavirus strains with altered VP4-VP7 combinations. The initial interaction of rotavirus strains with SA residues on the cell surface correlated with VP4 genotype specificity, not with species of origin or VP7 G serotype specificity (P = 0.001; r2 = 1.00, Pearson's correlation coefficient). In addition to being a requirement for infectivity, the presence of SA residues on the cell surface is a requirement for efficient growth in cell culture; recognition of the association of specific P genotypes with the binding of rotavirus to SA residues will facilitate our understanding of the molecular basis of the early events of rotavirus-cell interactions in cell culture models and of pathogenicity in vivo. PMID- 11907250 TI - Vesicular stomatitis virus glycoprotein does not determine the site of virus release in polarized epithelial cells. AB - In polarized epithelial cells, the vesicular stomatitis virus glycoprotein is segregated to the basolateral plasma membrane, where budding of the virus takes place. We have generated recombinant viruses expressing mutant glycoproteins without the basolateral-membrane-targeting signal in the cytoplasmic domain. Though about 50% of the mutant glycoproteins were found at the apical plasma membranes of infected MDCK cells, the virus was still predominantly released at the basolateral membranes, indicating that factors other than the glycoprotein determine the site of virus budding. PMID- 11907249 TI - Heat shock cognate protein 70 is involved in rotavirus cell entry. AB - In this work, we have identified the heat shock cognate protein (hsc70) as a receptor candidate for rotaviruses. hsc70 was shown to be present on the surface of MA104 cells, and antibodies to this protein blocked rotavirus infectivity, while not affecting the infectivity of reovirus and poliovirus. Preincubation of the hsc70 protein with the viruses also inhibited their infectivity. Triple layered particles (mature virions), but not double-layered particles, bound hsc70 in a solid-phase assay, and this interaction was blocked by monoclonal antibodies to the virus surface proteins VP4 and VP7. Rotaviruses were shown to interact with hsc70 at a postattachment step, since antibodies to hsc70 and the protein itself did not inhibit the virus attachment to cells. We propose that the functional rotavirus receptor is a complex of several cell surface molecules that include, among others, hsc70. PMID- 11907251 TI - Tat-vaccinated macaques do not control simian immunodeficiency virus SIVmac239 replication. AB - The regulatory proteins of human immunodeficiency virus may represent important vaccine targets. Here we assessed the role of Tat-specific cytotoxic T lymphocytes (CTL) in controlling pathogenic simian immunodeficiency virus SIVmac239 replication after using a DNA-prime, vaccinia virus Ankara-boost vaccine regimen. Despite the induction of Tat-specific CTL, there was no significant reduction in either peak or viral set point compared to that of controls. PMID- 11907252 TI - Nucleoprotein structure of immediate-early promoters Zp and Rp and of oriLyt of latent Epstein-Barr virus genomes. AB - Genomic footprints across Rp, Zp, and oriLyt of Epstein-Barr virus (EBV) have been conducted in a panel of latently infected B-cell lines. Close protein-base contacts were found about 360 nucleotides upstream of the Zp initiation site. Gel shifts and transient transfection assays indicated that an Sp1-NF1 locus may serve as a repressive transcriptional element against Zp induction from latent EBV genomes. PMID- 11907253 TI - Inter- and intramolecular recombinations in the cucumber mosaic virus genome related to adaptation to alstroemeria. AB - In four distinct alstroemeria-infecting cucumber mosaic virus (CMV) isolates, additional sequences of various lengths were present in the 3' nontranslated regions of their RNAs 2 and 3, apparently the result of intra- and intermolecular recombination events. Competition experiments revealed that these recombined RNA 2 and 3 segments increased the biological fitness of CMV in alstroemeria. PMID- 11907254 TI - Human immunodeficiency virus type 1 Vpr protein does not modulate surface expression of the CD4 receptor. AB - The CD4 receptor is required for the entry of human immunodeficiency virus (HIV) into target cells. It has long been known that Nef, Env, and Vpu participate in the removal of the viral receptor from the cell surface. Recently, it has been proposed that the HIV type 1 (HIV-1) Vpr protein may also play a role in the downmodulation of CD4 from the surfaces of infected cells (L. Conti, B. Varano, M. C. Gauzzi, P. Matarrese, M. Federico, W. Malorani, F. Belardelli, and S. Gessani, J. Virol. 74:10207-10211, 2000). To investigate the possible role of Vpr in the downregulation of the viral receptor Vpr alleles from HIV-1 and simian immunodeficiency virus were transiently expressed in transformed T cells and in 293T fibroblasts, and their ability to modulate surface CD4 was evaluated. All Vpr alleles efficiently arrested cells in the G(2) stage of the cell cycle. However, none of the tested Vpr proteins altered the expression of CD4 on the cell surface. In comparison, HIV-1 Nef efficiently downmodulated surface CD4 in all the experimental settings. Transformed T cells and primary lymphocytes were challenged with wild-type, Nef-defective, and Vpr-defective viruses. A significant reduction in the HIV-induced downmodulation of surface CD4 was observed in viruses lacking Nef. However, Vpr-deletion-containing viruses showed no defect in their ability to remove CD4 from the surfaces of infected cells. Our results indicate that Vpr does not play a role in the HIV-induced downmodulation of the CD4 receptor. PMID- 11907255 TI - Role of RNA in facilitating Gag/Gag-Pol interaction. AB - We have examined the influence of RNA upon the interaction of Gag-Pol with Gag during human immunodeficiency virus type 1 (HIV-1) assembly. COS7 cells were transfected with protease-negative HIV-1 proviral DNA, and Gag/Gag-Pol complexes were detected by coimmunoprecipitation with anti-integrase. In COS7 cells, Gag/Gag-Pol is found almost entirely in pelletable, membrane-bound complexes. Exposure of cells to 1% Triton X-100 releases Gag/Gag-Pol from bulk membrane, but the complexes remain pelletable. The role of RNA in facilitating the interaction between Gag and Gag-Pol was examined in these bulk membrane-free, pelletable complexes. The specific presence of viral genomic RNA is not required to maintain the Gag/Gag-Pol interaction, but some type of RNA is, since exposure to RNase destabilized the Gag/Gag-Pol complex. When present only in Gag, the nucleocapsid mutation R7R10K11S, which inhibits Gag binding to RNA, inhibits the formation of both Gag and Gag/Gag-Pol complexes. When present only in Gag-Pol, this mutation has no effect upon complex formation. This result indicates that Gag-Pol may not interact directly with RNA but rather requires RNA-facilitated Gag multimerization for its interaction with Gag. PMID- 11907256 TI - Intrinsic stability of episomal circles formed during human immunodeficiency virus type 1 replication. AB - The development of surrogate markers capable of detecting residual ongoing human immunodeficiency virus type 1 (HIV-1) replication in patients receiving highly active antiretroviral therapy is an important step in understanding viral dynamics and in developing new treatment strategies. In this study, we evaluated the utility of circular forms of the viral genome for the detection of recent infection of cells by HIV-1. We measured the fate of both one-long terminal repeat (1-LTR) and 2-LTR circles following in vitro infection of logarithmically growing CD4+ T cells under conditions in which cell death was not a significant contributing factor. Circular forms of the viral genome were found to be highly stable and to decrease in concentration only as a function of dilution resulting from cell division. We conclude that these DNA circles are not intrinsically unstable in all cell types and suggest that the utility of 2-LTR circle assays in measuring recent HIV-1 infection of susceptible cells in vivo needs to be reevaluated. PMID- 11907257 TI - Identification of human herpesvirus 6 latency-associated transcripts. AB - Four kinds of latency-associated transcripts of human herpesvirus 6 were identified which were detected only in latently infected cells. Although they were oriented in the same direction as the immediate-early 1 and 2 (IE1/IE2) genes and shared their protein-coding region with IE1/IE2, their transcription start sites and exon(s) were latency associated. PMID- 11907258 TI - Genetic ablation of phosphatidylinositol transfer protein function in murine embryonic stem cells. AB - Phosphatidylinositol transfer proteins (PITPs) regulate the interface between signal transduction, membrane-trafficking, and lipid metabolic pathways in eukaryotic cells. The best characterized mammalian PITPs are PITP alpha and PITP beta, two highly homologous proteins that are encoded by distinct genes. Insights into PITP alpha and PITP beta function in mammalian systems have been gleaned exclusively from cell-free or permeabilized cell reconstitution and resolution studies. Herein, we report for the first time the use of genetic approaches to directly address the physiological functions of PITP alpha and PITP beta in murine cells. Contrary to expectations, we find that ablation of PITP alpha function in murine cells fails to compromise growth and has no significant consequence for bulk phospholipid metabolism. Moreover, the data show that PITP alpha does not play an obvious role in any of the cellular activities where it has been reconstituted as an essential stimulatory factor. These activities include protein trafficking through the constitutive secretory pathway, endocytic pathway function, biogenesis of mast cell dense core secretory granules, and the agonist-induced fusion of dense core secretory granules to the mast cell plasma membrane. Finally, the data demonstrate that PITP alpha-deficient cells not only retain their responsiveness to bulk growth factor stimulation but also retain their pluripotency. In contrast, we were unable to evict both PITP beta alleles from murine cells and show that PITP beta deficiency results in catastrophic failure early in murine embryonic development. We suggest that PITP beta is an essential housekeeping PITP in murine cells, whereas PITP alpha plays a far more specialized function in mammals than that indicated by in vitro systems that show PITP dependence. PMID- 11907259 TI - Checkpoint protein BubR1 acts synergistically with Mad2 to inhibit anaphase promoting complex. AB - The spindle assembly checkpoint monitors the attachment of kinetochores to the mitotic spindle and the tension exerted on kinetochores by microtubules and delays the onset of anaphase until all the chromosomes are aligned at the metaphase plate. The target of the checkpoint control is the anaphase-promoting complex (APC)/cyclosome, a ubiquitin ligase whose activation by Cdc20 is required for separation of sister chromatids. In response to activation of the checkpoint, Mad2 binds to and inhibits Cdc20-APC. I show herein that in checkpoint-arrested cells, human Cdc20 forms two separate, inactive complexes, a lower affinity complex with Mad2 and a higher affinity complex with BubR1. Purified BubR1 binds to recombinant Cdc20 and this interaction is direct. Binding of BubR1 to Cdc20 inhibits activation of APC and this inhibition is independent of its kinase activity. Quantitative analysis indicates that BubR1 is 12-fold more potent than Mad2 as an inhibitor of Cdc20. Although at high protein concentrations BubR1 and Mad2 each is sufficient to inhibit Cdc20, BubR1 and Mad2 mutually promote each other's binding to Cdc20 and function synergistically at physiological concentrations to quantitatively inhibit Cdc20-APC. Thus, BubR1 and Mad2 act cooperatively to prevent premature separation of sister chromatids by directly inhibiting APC. PMID- 11907260 TI - Palmitoylation of tetraspanin proteins: modulation of CD151 lateral interactions, subcellular distribution, and integrin-dependent cell morphology. AB - Here we demonstrate that multiple tetraspanin (transmembrane 4 superfamily) proteins are palmitoylated, in either the Golgi or a post-Golgi compartment. Using CD151 as a model tetraspanin, we identified and mutated intracellular N terminal and C-terminal cysteine palmitoylation sites. Simultaneous mutations of C11, C15, C242, and C243 (each to serine) eliminated >90% of CD151 palmitoylation. Notably, palmitoylation had minimal influence on the density of tetraspanin protein complexes, did not promote tetraspanin localization into detergent-resistant microdomains, and was not required for CD151-alpha 3 beta 1 integrin association. However, the CD151 tetra mutant showed markedly diminished associations with other cell surface proteins, including other transmembrane 4 superfamily proteins (CD9, CD63). Thus, palmitoylation may be critical for assembly of the large network of cell surface tetraspanin-protein interactions, sometimes called the "tetraspanin web." Also, compared with wild-type CD151, the tetra mutant was much more diffusely distributed and showed markedly diminished stability during biosynthesis. Finally, expression of the tetra-CD151 mutant profoundly altered alpha 3 integrin-deficient kidney epithelial cells, such that they converted from a dispersed, elongated morphology to an epithelium-like cobblestone clustering. These results point to novel biochemical and biological functions for tetraspanin palmitoylation. PMID- 11907261 TI - Genomic analysis of homotypic vacuole fusion. AB - Yeast vacuoles undergo fission and homotypic fusion, yielding one to three vacuoles per cell at steady state. Defects in vacuole fusion result in vacuole fragmentation. We have screened 4828 yeast strains, each with a deletion of a nonessential gene, for vacuole morphology defects. Fragmented vacuoles were found in strains deleted for genes encoding known fusion catalysts as well as 19 enzymes of lipid metabolism, 4 SNAREs, 12 GTPases and GTPase effectors, 9 additional known vacuole protein-sorting genes, 16 protein kinases, 2 phosphatases, 11 cytoskeletal proteins, and 28 genes of unknown function. Vacuole fusion and vacuole protein sorting are catalyzed by distinct, but overlapping, sets of proteins. Novel pathways of vacuole priming and docking emerged from this deletion screen. These include ergosterol biosynthesis, phosphatidylinositol (4,5)-bisphosphate turnover, and signaling from Rho GTPases to actin remodeling. These pathways are supported by the sensitivity of the late stages of vacuole fusion to inhibitors of phospholipase C, calcium channels, and actin remodeling. Using databases of yeast protein interactions, we found that many nonessential genes identified in our deletion screen interact with essential genes that are directly involved in vacuole fusion. Our screen reveals regulatory pathways of vacuole docking and provides a genomic basis for studies of this reaction. PMID- 11907262 TI - RTG-dependent mitochondria-to-nucleus signaling is regulated by MKS1 and is linked to formation of yeast prion [URE3]. AB - An important function of the RTG signaling pathway is maintenance of intracellular glutamate supplies in yeast cells with dysfunctional mitochondria. Herein, we report that MKS1 is a negative regulator of the RTG pathway, acting between Rtg2p, a proximal sensor of mitochondrial function, and the bHLH transcription factors Rtg1p and Rtg3p. In mks1 Delta cells, RTG target gene expression is constitutive, bypassing the requirement for Rtg2p, and is no longer repressible by glutamate. We show further that Mks1p is a phosphoprotein whose phosphorylation pattern parallels that of Rtg3p in response to activation of the RTG pathway, and that Mks1p is in a complex with Rtg2p. MKS1 was previously implicated in the formation of [URE3], an inactive prion form of a negative regulator of the nitrogen catabolite repression pathway, Ure2p. rtg Delta mutations induce [URE3] and can do so independently of MKS1. We find that glutamate suppresses [URE3] formation, suggesting that the Mks1p effect on the formation of [URE3] can occur indirectly via regulation of the RTG pathway. PMID- 11907263 TI - Distinct regulatory proteins control the graded transcriptional response to increasing H(2)O(2) levels in fission yeast Schizosaccharomyces pombe. AB - The signaling pathways that sense adverse stimuli and communicate with the nucleus to initiate appropriate changes in gene expression are central to the cellular stress response. Herein, we have characterized the role of the Sty1 (Spc1) stress-activated mitogen-activated protein kinase pathway, and the Pap1 and Atf1 transcription factors, in regulating the response to H(2)O(2) in the fission yeast Schizosaccharomyces pombe. We find that H(2)O(2) activates the Sty1 pathway in a dose-dependent manner via at least two sensing mechanisms. At relatively low levels of H(2)O(2), a two component-signaling pathway, which feeds into either of the two stress-activated mitogen-activated protein kinase kinase kinases Wak1 or Win1, regulates Sty1 phosphorylation. In contrast, at high levels of H(2)O(2), Sty1 activation is controlled predominantly by a two-component independent mechanism and requires the function of both Wak1 and Win1. Individual transcription factors were also found to function within a limited range of H(2)O(2) concentrations. Pap1 activates target genes primarily in response to low levels of H(2)O(2), whereas Atf1 primarily controls the transcriptional response to high concentrations of H(2)O(2). Our results demonstrate that S. pombe uses a combination of stress-responsive regulatory proteins to gauge and effect the appropriate transcriptional response to increasing concentrations of H(2)O(2). PMID- 11907264 TI - Identification of a novel light intermediate chain (D2LIC) for mammalian cytoplasmic dynein 2. AB - The diversity of dynein's functions in mammalian cells is a manifestation of both the existence of multiple dynein heavy chain isoforms and an extensive set of associated protein subunits. In this study, we have identified and characterized a novel subunit of the mammalian cytoplasmic dynein 2 complex. The sequence similarity between this 33-kDa subunit and the light intermediate chains (LICs) of cytoplasmic dynein 1 suggests that this protein is a dynein 2 LIC (D2LIC). D2LIC contains a P-loop motif near its NH(2) terminus, and it shares a short region of similarity to the yeast GTPases Spg1p and Tem1p. The D2LIC subunit interacts specifically with DHC2 (or cDhc1b) in both reciprocal immunoprecipitations and sedimentation assays. The expression of D2LIC also mirrors that of DHC2 in a variety of tissues. D2LIC colocalizes with DHC2 at the Golgi apparatus throughout the cell cycle. On brefeldin A-induced Golgi fragmentation, a fraction of D2LIC redistributes to the cytoplasm, leaving behind a subset of D2LIC that is localized around the centrosome. Our results suggest that D2LIC is a bona fide subunit of cytoplasmic dynein 2 that may play a role in maintaining Golgi organization by binding cytoplasmic dynein 2 to its Golgi associated cargo. PMID- 11907265 TI - Stretch-regulated exocytosis/endocytosis in bladder umbrella cells. AB - The epithelium of the urinary bladder must maintain a highly impermeable barrier despite large variations in urine volume during bladder filling and voiding. To study how the epithelium accommodates these volume changes, we mounted bladder tissue in modified Ussing chambers and subjected the tissue to mechanical stretch. Stretching the tissue for 5 h resulted in a 50% increase in lumenal surface area (from approximately 2900 to 4300 microm(2)), exocytosis of a population of discoidal vesicles located in the apical cytoplasm of the superficial umbrella cells, and release of secretory proteins. Surprisingly, stretch also induced endocytosis of apical membrane and 100% of biotin-labeled membrane was internalized within 5 min after stretch. The endocytosed membrane was delivered to lysosomes and degraded by a leupeptin-sensitive pathway. Last, we show that the exocytic events were mediated, in part, by a cyclic adenosine monophosphate, protein kinase A-dependent process. Our results indicate that stretch modulates mucosal surface area by coordinating both exocytosis and endocytosis at the apical membrane of umbrella cells and provide insight into the mechanism of how mechanical forces regulate membrane traffic in non-excitable cells. PMID- 11907266 TI - Genetic basis of mitochondrial function and morphology in Saccharomyces cerevisiae. AB - The understanding of the processes underlying organellar function and inheritance requires the identification and characterization of the molecular components involved. We pursued a genomic approach to define the complements of genes required for respiratory growth and inheritance of mitochondria with normal morphology in yeast. With the systematic screening of a deletion mutant library covering the nonessential genes of Saccharomyces cerevisiae the numbers of genes known to be required for respiratory function and establishment of wild-type-like mitochondrial structure have been more than doubled. In addition to the identification of novel components, the systematic screen revealed unprecedented mitochondrial phenotypes that have never been observed by conventional screens. These data provide a comprehensive picture of the cellular processes and molecular components required for mitochondrial function and structure in a simple eukaryotic cell. PMID- 11907267 TI - Endoplasmic reticulum dynamics, inheritance, and cytoskeletal interactions in budding yeast. AB - The endoplasmic reticulum (ER) in Saccharomyces cerevisiae consists of a reticulum underlying the plasma membrane (cortical ER) and ER associated with the nuclear envelope (nuclear ER). We used a Sec63p-green fluorescent protein fusion protein to study motility events associated with inheritance of cortical ER and nuclear ER in living yeast cells. During M phase before nuclear migration, we observed thick, apparently rigid tubular extensions emanating from the nuclear ER that elongate, undergo sweeping motions along the cell cortex, and shorten. Two findings support a role for microtubules in this process. First, extension of tubular structures from the nuclear ER is inhibited by destabilization of microtubules. Second, astral microtubules, structures that undergo similar patterns of extension, cortical surveillance and retraction, colocalize with nuclear ER extensions. During S and G(2) phases of the cell cycle, we observed anchorage of the cortical ER at the site of bud emergence and apical bud growth. Thin tubules of the ER that extend from the anchored cortical ER display undulating, apparently random movement and move into the bud as it grows. Finally, we found that cortical ER morphology is sensitive to a filamentous actin destabilizing drug, latrunculin-A, and to mutations in the actin-encoding ACT1 gene. Our observations support 1) different mechanisms and cytoskeletal mediators for the inheritance of nuclear and cortical ER elements and 2) a mechanism for cortical ER inheritance that is cytoskeleton dependent but relies on anchorage, not directed movement. PMID- 11907269 TI - Erv14p directs a transmembrane secretory protein into COPII-coated transport vesicles. AB - Erv14p is a conserved integral membrane protein that traffics in COPII-coated vesicles and localizes to the early secretory pathway in yeast. Deletion of ERV14 causes a defect in polarized growth because Axl2p, a transmembrane secretory protein, accumulates in the endoplasmic reticulum and is not delivered to its site of function on the cell surface. Herein, we show that Erv14p is required for selection of Axl2p into COPII vesicles and for efficient formation of these vesicles. Erv14p binds to subunits of the COPII coat and binding depends on conserved residues in a cytoplasmically exposed loop domain of Erv14p. When mutations are introduced into this loop, an Erv14p-Axl2p complex accumulates in the endoplasmic reticulum, suggesting that Erv14p links Axl2p to the COPII coat. Based on these results and further genetic experiments, we propose Erv14p coordinates COPII vesicle formation with incorporation of specific secretory cargo. PMID- 11907268 TI - Regulation of protein transport from the Golgi complex to the endoplasmic reticulum by CDC42 and N-WASP. AB - Actin is involved in the organization of the Golgi complex and Golgi-to-ER protein transport in mammalian cells. Little, however, is known about the regulation of the Golgi-associated actin cytoskeleton. We provide evidence that Cdc42, a small GTPase that regulates actin dynamics, controls Golgi-to-ER protein transport. We located GFP-Cdc42 in the lateral portions of Golgi cisternae and in COPI-coated and non-coated Golgi-associated transport intermediates. Overexpression of Cdc42 and its activated form Cdc42V12 inhibited the retrograde transport of Shiga toxin from the Golgi complex to the ER, the redistribution of the KDEL receptor, and the ER accumulation of Golgi-resident proteins induced by the active GTP-bound mutant of Sar1 (Sar1[H79G]). Coexpression of wild-type or activated Cdc42 and N-WASP also inhibited Golgi-to-ER transport, but this was not the case in cells expressing Cdc42V12 and N-WASP(Delta WA), a mutant form of N WASP that lacks Arp2/3 binding. Furthermore, Cdc42V12 recruited GFP-N-WASP to the Golgi complex. We therefore conclude that Cdc42 regulates Golgi-to-ER protein transport in an N-WASP-dependent manner. PMID- 11907270 TI - Lamin-dependent localization of UNC-84, a protein required for nuclear migration in Caenorhabditis elegans. AB - Mutations in the Caenorhabditis elegans unc-84 gene cause defects in nuclear migration and anchoring. We show that endogenous UNC-84 protein colocalizes with Ce-lamin at the nuclear envelope and that the envelope localization of UNC-84 requires Ce-lamin. We also show that during mitosis, UNC-84 remains at the nuclear periphery until late anaphase, similar to known inner nuclear membrane proteins. UNC-84 protein is first detected at the 26-cell stage and thereafter is present in most cells during development and in adults. UNC-84 is properly expressed in unc-83 and anc-1 lines, which have phenotypes similar to unc-84, suggesting that neither the expression nor nuclear envelope localization of UNC 84 depends on UNC-83 or ANC-1 proteins. The envelope localization of Ce-lamin, Ce emerin, Ce-MAN1, and nucleoporins are unaffected by the loss of UNC-84. UNC-84 is not required for centrosome attachment to the nucleus because centrosomes are localized normally in unc-84 hyp7 cells despite a nuclear migration defect. Models for UNC-84 localization are discussed. PMID- 11907271 TI - Transforming growth factor-beta-induced mobilization of actin cytoskeleton requires signaling by small GTPases Cdc42 and RhoA. AB - Transforming growth factor-beta (TGF-beta) is a potent regulator of cell growth and differentiation in many cell types. The Smad signaling pathway constitutes a main signal transduction route downstream of TGF-beta receptors. We studied TGF beta-induced rearrangements of the actin filament system and found that TGF-beta 1 treatment of PC-3U human prostate carcinoma cells resulted in a rapid formation of lamellipodia. Interestingly, this response was shown to be independent of the Smad signaling pathway; instead, it required the activity of the Rho GTPases Cdc42 and RhoA, because ectopic expression of dominant negative mutant Cdc42 and RhoA abrogated the response. Long-term stimulation with TGF-beta 1 resulted in an assembly of stress fibers; this response required both signaling via Cdc42 and RhoA, and Smad proteins. A known downstream effector of Cdc42 is p38(MAPK); treatment of the cells with the p38(MAPK) inhibitor 4-(4-fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(pyridyl)1H-imidazole (SB203580), as well as ectopic expression of a kinase-inactive p38(MAPK), abrogated the TGF-beta-induced actin reorganization. Moreover, treatment of cells with the inhibitors of the RhoA target-protein Rho-associated coiled-coil kinase (+)-R-trans-4-(aminoethyl)-N-(4 pyridyl) cyclohexanecarboxamide (Y-27632) and 1-5( isoquinolinesulfonyl)homopiperazine (HA-1077), as well as ectopic expression of kinase-inactive Rho coiled-coil kinase-1, abrogated the TGF-beta 1-induced formation of stress fibers. Collectively, these data indicate that TGF-beta induced membrane ruffles occur via Rho GTPase-dependent pathways, whereas long term effects require cooperation between Smad and Rho GTPase signaling pathways. PMID- 11907272 TI - FBI-1 can stimulate HIV-1 Tat activity and is targeted to a novel subnuclear domain that includes the Tat-P-TEFb-containing nuclear speckles. AB - FBI-1 is a cellular POZ-domain-containing protein that binds to the HIV-1 LTR and associates with the HIV-1 transactivator protein Tat. Here we show that elevated levels of FBI-1 specifically stimulate Tat activity and that this effect is dependent on the same domain of FBI-1 that mediates Tat-FBI-1 association in vivo. FBI-1 also partially colocalizes with Tat and Tat's cellular cofactor, P TEFb (Cdk9 and cyclin T1), at the splicing-factor-rich nuclear speckle domain. Further, a less-soluble population of FBI-1 distributes in a novel peripheral speckle pattern of localization as well as in other nuclear regions. This distribution pattern is dependent on the FBI-1 DNA binding domain, on the presence of cellular DNA, and on active transcription. Taken together, these results suggest that FBI-1 is a cellular factor that preferentially associates with active chromatin and that can specifically stimulate Tat-activated HIV-1 transcription. PMID- 11907273 TI - The 14-kDa dynein light chain-family protein Dlc1 is required for regular oscillatory nuclear movement and efficient recombination during meiotic prophase in fission yeast. AB - A Schizosaccharomyces pombe spindle pole body (SPB) protein interacts in a two hybrid system with Dlc1, which belongs to the 14-kDa Tctex-1 dynein light chain family. Green fluorescent protein-tagged Dlc1 accumulated at the SPB throughout the life cycle. During meiotic prophase, Dlc1 was present along astral microtubules and microtubule-anchoring sites on the cell cortex, reminiscent of the cytoplasmic dynein heavy chain Dhc1. In a dlc1-null mutant, Dhc1-dependent nuclear movement in meiotic prophase became irregular in its duration and direction. Dhc1 protein was displaced from the cortex anchors and the formation of microtubule bundle(s) that guide nuclear movement was impaired in the mutant. Meiotic recombination in the dlc1 mutant was reduced to levels similar to that in the dhc1 mutant. Dlc1 and Dhc1 also have roles in karyogamy and rDNA relocation during the sexual phase. Strains mutated in both the dlc1 and dhc1 loci displayed more severe defects in recombination, karyogamy, and sporulation than in either single mutant alone, suggesting that Dlc1 is involved in nuclear events that are independent of Dhc1. S. pombe contains a homolog of the 8-kDa dynein light chain, Dlc2. This class of dynein light chain, however, is not essential in either the vegetative or sexual phases. PMID- 11907275 TI - Dynein supports motility of endoplasmic reticulum in the fungus Ustilago maydis. AB - The endoplasmic reticulum (ER) of most vertebrate cells is spread out by kinesin dependent transport along microtubules, whereas studies in Saccharomyces cerevisiae indicated that motility of fungal ER is an actin-based process. However, microtubules are of minor importance for organelle transport in yeast, but they are crucial for intracellular transport within numerous other fungi. Herein, we set out to elucidate the role of the tubulin cytoskeleton in ER organization and dynamics in the fungal pathogen Ustilago maydis. An ER-resident green fluorescent protein (GFP)-fusion protein localized to a peripheral network and the nuclear envelope. Tubules and patches within the network exhibited rapid dynein-driven motion along microtubules, whereas conventional kinesin did not participate in ER motility. Cortical ER organization was independent of microtubules or F-actin, but reformation of the network after experimental disruption was mediated by microtubules and dynein. In addition, a polar gradient of motile ER-GFP stained dots was detected that accumulated around the apical Golgi apparatus. Both the gradient and the Golgi apparatus were sensitive to brefeldin A or benomyl treatment, suggesting that the gradient represents microtubule-dependent vesicle trafficking between ER and Golgi. Our results demonstrate a role of cytoplasmic dynein and microtubules in motility, but not peripheral localization of the ER in U. maydis. PMID- 11907274 TI - Proliferating or differentiating stimuli act on different lipid-dependent signaling pathways in nuclei of human leukemia cells. AB - Previous results have shown that the human promyelocytic leukemia HL-60 cell line responds to either proliferating or differentiating stimuli. When these cells are induced to proliferate, protein kinase C (PKC)-beta II migrates toward the nucleus, whereas when they are exposed to differentiating agents, there is a nuclear translocation of the alpha isoform of PKC. As a step toward the elucidation of the early intranuclear events that regulate the proliferation or the differentiation process, we show that in the HL-60 cells, a proliferating stimulus (i.e., insulin-like growth factor-I [IGF-I]) increased nuclear diacylglycerol (DAG) production derived from phosphatidylinositol (4,5) bisphosphate, as indicated by the inhibition exerted by 1-O-octadeyl-2-O-methyl sn-glycero-3-phosphocholine and U-73122 (1-[6((17 beta-3-methoxyestra-1,3,5(10) trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione), which are pharmacological inhibitors of phosphoinositide-specific phospholipase C. In contrast, when HL-60 cells were induced to differentiate along the granulocytic lineage by dimethyl sulfoxide, we observed a rise in the nuclear DAG mass, which was sensitive to either neomycin or propranolol, two compounds with inhibitory effect on phospholipase D (PLD)-mediated DAG generation. In nuclei of dimethyl sulfoxide treated HL-60 cells, we observed a rise in the amount of a 90-kDa PLD, distinct from PLD1 or PLD2. When a phosphatidylinositol (4,5) bisphosphate-derived DAG pool was generated in the nucleus, a selective translocation of PKC-beta II occurred. On the other hand, nuclear DAG derived through PLD, recruited PKC-alpha to the nucleus. Both of these PKC isoforms were phosphorylated on serine residues. These results provide support for the proposal that in the HL-60 cell nucleus there are two independently regulated sources of DAG, both of which are capable of acting as the driving force that attracts to this organelle distinct, DAG-dependent PKC isozymes. Our results assume a particular significance in light of the proposed use of pharmacological inhibitors of PKC-dependent biochemical pathways for the therapy of cancer disease. PMID- 11907276 TI - Activation and caspase-mediated inhibition of PARP: a molecular switch between fibroblast necrosis and apoptosis in death receptor signaling. AB - Death ligands not only induce apoptosis but can also trigger necrosis with distinct biochemical and morphological features. We recently showed that in L929 cells CD95 ligation induces apoptosis, whereas TNF elicits necrosis. Treatment with anti-CD95 resulted in typical apoptosis characterized by caspase activation and DNA fragmentation. These events were barely induced by TNF, although TNF triggered cell death to a similar extent as CD95. Surprisingly, whereas the caspase inhibitor zVAD prevented CD95-mediated apoptosis, it potentiated TNF induced necrosis. Cotreatment with TNF and zVAD was characterized by ATP depletion and accelerated necrosis. To investigate the mechanisms underlying TNF induced cell death and its potentiation by zVAD, we examined the role of poly(ADP ribose)polymerase-1 (PARP-1). TNF but not CD95 mediated PARP activation, whereas a PARP inhibitor suppressed TNF-induced necrosis and the sensitizing effect of zVAD. In addition, fibroblasts expressing a noncleavable PARP-1 mutant were more sensitive to TNF than wild-type cells. Our results indicate that TNF induces PARP activation leading to ATP depletion and subsequent necrosis. In contrast, in CD95 mediated apoptosis caspases cause PARP-1 cleavage and thereby maintain ATP levels. Because ATP is required for apoptosis, we suggest that PARP-1 cleavage functions as a molecular switch between apoptotic and necrotic modes of death receptor-induced cell death. PMID- 11907277 TI - The localization of the integral membrane protein Cps1p to the cell division site is dependent on the actomyosin ring and the septation-inducing network in Schizosaccharomyces pombe. AB - Schizosaccharomyces pombe cells divide by medial fission through the use of an actomyosin-based contractile ring. Constriction of the actomyosin ring is accompanied by the centripetal addition of new membranes and cell wall material. In this article, we characterize the mechanism responsible for the localization of Cps1p, a septum-synthesizing 1,3-beta-glucan synthase, to the division site during cytokinesis. We show that Cps1p is an integral membrane protein that localizes to the cell division site late in anaphase. Neither F-actin nor microtubules are essential for the initial assembly of Cps1p to the medial division site. F-actin, but not microtubules, is however important for the eventual incorporation of Cps1p into the actomyosin ring. Assembly of Cps1p into the cell division ring is also dependent on the septation-inducing network (SIN) proteins that regulate division septum formation after assembly of the actomyosin ring. Fluorescence-recovery after-photobleaching experiments reveal that Cps1p does not diffuse appreciably within the plasma membrane and is retained at the division site by a mechanism that does not depend on an intact F-actin cytoskeleton. We conclude that the actomyosin ring serves as a spatial cue for Cps1p localization, whereas the maintenance of Cps1p at the division site occurs by a novel F-actin- and microtubule-independent mechanism. Furthermore, we propose that the SIN proteins ensure localization of Cps1p at the appropriate point in the cell cycle. PMID- 11907278 TI - Wound closure in the lamellipodia of single cells: mediation by actin polymerization in the absence of an actomyosin purse string. AB - The actomyosin purse string is an evolutionarily conserved contractile structure that is involved in cytokinesis, morphogenesis, and wound healing. Recent studies suggested that an actomyosin purse string is crucial for the closure of wounds in single cells. In the present study, morphological and pharmacological methods were used to investigate the role of this structure in the closure of wounds in the peripheral cytoplasm of sea urchin coelomocytes. These discoidal shaped cells underwent a dramatic form of actin-based centripetal/retrograde flow and occasionally opened and closed spontaneous wounds in their lamellipodia. Fluorescent phalloidin staining indicated that a well defined fringe of actin filaments assembles from the margin of these holes, and drug studies with cytochalasin D and latrunculin A indicated that actin polymerization is required for wound closure. Additional evidence that actin polymerization is involved in wound closure was provided by the localization of components of the Arp2/3 complex to the wound margin. Significantly, myosin II immunolocalization demonstrated that it is not associated with wound margins despite being present in the perinuclear region. Pharmacological evidence for the lack of myosin II involvement in wound closure comes from experiments in which a microneedle was used to produce wounds in cells in which actomyosin contraction was inhibited by treatment with kinase inhibitors. Wounds produced in kinase inhibitor-treated cells closed in a manner similar to that seen with control cells. Taken together, our results suggest that an actomyosin purse string mechanism is not responsible for the closure of lamellar wounds in coelomocytes. We hypothesize that the wounds heal by means of a combination of the force produced by actin polymerization alone and centripetal flow. Interestingly, these cells did assemble an actomyosin structure around the margin of phagosome-like membrane invaginations, indicating that myosin is not simply excluded from the periphery by some general mechanism. The results indicate that the actomyosin purse string is not the only mechanism that can mediate wound closure in single cells. PMID- 11907279 TI - The outer dynein arm-docking complex: composition and characterization of a subunit (oda1) necessary for outer arm assembly. AB - To learn more about how dyneins are targeted to specific sites in the flagellum, we have investigated a factor necessary for binding of outer arm dynein to the axonemal microtubules of Chlamydomonas. This factor, termed the outer dynein arm docking complex (ODA-DC), previously was shown to be missing from axonemes of the outer dynein armless mutants oda1 and oda3. We have now partially purified the ODA-DC, determined that it contains equimolar amounts of M(r) approximately 105,000 and approximately 70,000 proteins plus a third protein of M(r) approximately 25,000, and found that it is associated with the isolated outer arm in a 1:1 molar ratio. We have cloned a full-length cDNA encoding the M(r) approximately 70,000 protein; the sequence predicts a 62.5-kDa protein with potential homologs in higher ciliated organisms, including humans. Sequencing of corresponding cDNA from strain oda1 revealed it has a mutation resulting in a stop codon just downstream of the initiator ATG; thus, it is unable to make the full-length M(r) approximately 70,000 protein. These results demonstrate that the ODA1 gene encodes the M(r) approximately 70,000 protein, and that the protein is essential for assembly of the ODA-DC and the outer dynein arm onto the doublet microtubule. PMID- 11907281 TI - Association of a nonmuscle myosin II with axoplasmic organelles. AB - Association of motor proteins with organelles is required for the motors to mediate transport. Because axoplasmic organelles move on actin filaments, they must have associated actin-based motors, most likely members of the myosin superfamily. To gain a better understanding of the roles of myosins in the axon we used the giant axon of the squid, a powerful model for studies of axonal physiology. First, a approximately 220 kDa protein was purified from squid optic lobe, using a biochemical protocol designed to isolate myosins. Peptide sequence analysis, followed by cloning and sequencing of the full-length cDNA, identified this approximately 220 kDa protein as a nonmuscle myosin II. This myosin is also present in axoplasm, as determined by two independent criteria. First, RT-PCR using sequence-specific primers detected the transcript in the stellate ganglion, which contains the cell bodies that give rise to the giant axon. Second, Western blot analysis using nonmuscle myosin II isotype-specific antibodies detected a single approximately 220 kDa band in axoplasm. Axoplasm was fractionated through a four-step sucrose gradient after 0.6 M KI treatment, which separates organelles from cytoskeletal components. Of the total nonmuscle myosin II in axoplasm, 43.2% copurified with organelles in the 15% sucrose fraction, while the remainder (56.8%) was soluble and found in the supernatant. This myosin decorates the cytoplasmic surface of 21% of the axoplasmic organelles, as demonstrated by immunogold electron-microscopy. Thus, nonmuscle myosin II is synthesized in the cell bodies of the giant axon, is present in the axon, and is associated with isolated axoplasmic organelles. Therefore, in addition to myosin V, this myosin is likely to be an axoplasmic organelle motor. PMID- 11907280 TI - Cyclin A- and cyclin E-Cdk complexes shuttle between the nucleus and the cytoplasm. AB - Cyclins A and E and their partner cyclin-dependent kinases (Cdks) are key regulators of DNA synthesis and of mitosis. Immunofluorescence studies have shown that both cyclins are nuclear and that a proportion of cyclin A is localized to sites of DNA replication. However, recently, both cyclin A and cyclin E have been implicated as regulators of centrosome replication, and it is unclear when and where these cyclin-Cdks can interact with cytoplasmic substrates. We have used live cell imaging to study the behavior of cyclin/Cdk complexes. We found that cyclin A and cyclin E are able to regulate both nuclear and cytoplasmic events because they both shuttle between the nucleus and the cytoplasm. However, we found that there are marked differences in their shuttling behavior, which raises the possibility that cyclin/Cdk function could be regulated at the level of nuclear import and export. In the course of these experiments, we have also found that, contrary to published results, mutations in the hydrophobic patch of cyclin A do affect Cdk binding and nuclear import. This has implications for the role of the hydrophobic patch as a substrate selection motif. PMID- 11907282 TI - Two related subpellicular cytoskeleton-associated proteins in Trypanosoma brucei stabilize microtubules. AB - The subpellicular microtubules of the trypanosome cytoskeleton are cross-linked to each other and the plasma membrane, creating a cage-like structure. We have isolated, from Trypanosoma brucei, two related low-molecular-weight cytoskeleton associated proteins (15- and 17-kDa), called CAP15 and CAP17, which are differentially expressed during the life cycle. Immunolabeling shows a corset like colocalization of both CAPs and tubulin. Western blot and electron microscope analyses show CAP15 and CAP17 labeling on detergent-extracted cytoskeletons. However, the localization of both proteins is restricted to the anterior, microtubule minus, and less dynamic half of the corset. CAP15 and CAP17 share properties of microtubule-associated proteins when expressed in heterologous cells (Chinese hamster ovary and HeLa), colocalization with their microtubules, induction of microtubule bundle formation, cold resistance, and insensitivity to nocodazole. When overexpressed in T. brucei, both CAP15 and CAP17 cover the whole subpellicular corset and induce morphological disorders, cell cycle-based abnormalities, and subsequent asymmetric cytokinesis. PMID- 11907283 TI - Role of adaptor complex AP-3 in targeting wild-type and mutated CD63 to lysosomes. AB - CD63 is a lysosomal membrane protein that belongs to the tetraspanin family. Its carboxyterminal cytoplasmic tail sequence contains the lysosomal targeting motif GYEVM. Strong, tyrosine-dependent interaction of the wild-type carboxyterminal tail of CD63 with the AP-3 adaptor subunit mu 3 was observed using a yeast two hybrid system. The strength of interaction of mutated tail sequences with mu 3 correlated with the degree of lysosomal localization of similarly mutated human CD63 molecules in stably transfected normal rat kidney cells. Mutated CD63 containing the cytosolic tail sequence GYEVI, which interacted strongly with mu 3 but not at all with mu 2 in the yeast two-hybrid system, localized to lysosomes in transfected normal rat kidney and NIH-3T3 cells. In contrast, it localized to the cell surface in transfected cells of pearl and mocha mice, which have genetic defects in genes encoding subunits of AP-3, but to lysosomes in functionally rescued mocha cells expressing the delta subunit of AP-3. Thus, AP-3 is absolutely required for the delivery of this mutated CD63 to lysosomes. Using this AP-3-dependent mutant of CD63, we have shown that AP-3 functions in membrane traffic from the trans-Golgi network to lysosomes via an intracellular route that appears to bypass early endosomes. PMID- 11907284 TI - Protein phosphatases 1 and 2A transiently associate with myosin during the peak rate of secretion from mast cells. AB - Mast cells undergo cytoskeletal restructuring to allow secretory granules passage through the cortical actomyosin barrier to fuse with the plasma membrane and release inflammatory mediators. Protein phosphorylation is believed to regulate these rearrangements. Although some of the protein kinases implicated in this phosphorylation are known, the relevant protein phosphatases are not. At the peak rate of antigen-induced granule mediator release (2.5 min), protein phosphatases PP1 and PP2A, along with actin and myosin II, are transiently relocated to ruffles on the apical surface and a band at the peripheral edge of the cell. This leaves an area between the nucleus and the peripheral edge significantly depleted (3-5-fold) in these proteins. Phorbol 12-myristate 13-acetate (PMA) plus A23187 induces the same changes, at a time coincident with its slower rate of secretion. Coimmunoprecipitation experiments demonstrated a significantly increased association of myosin with PP1 and PP2A at the time of peak mediator release, with levels of association decreasing by 5 min. Jasplakinolide, an inhibitor of actin assembly, inhibits secretion and the cytoskeletal rearrangements. Surprisingly, jasplakinolide also affects myosin, inducing the formation of short rods throughout the cytoplasm. Inhibition of PP2A inhibited secretion, the cytoskeletal rearrangements, and led to increased phosphorylation of the myosin heavy and light chains at protein kinase C-specific sites. These findings indicate that a dynamic actomyosin cytoskeleton, partially regulated by both PP1 and PP2A, is required for mast cell secretion. PMID- 11907285 TI - Specialized care for elderly patients. PMID- 11907286 TI - Prophylactic implantation of a defibrillator in patients with myocardial infarction and reduced ejection fraction. AB - BACKGROUND: Patients with reduced left ventricular function after myocardial infarction are at risk for life-threatening ventricular arrhythmias. This randomized trial was designed to evaluate the effect of an implantable defibrillator on survival in such patients. METHODS: Over the course of four years, we enrolled 1232 patients with a prior myocardial infarction and a left ventricular ejection fraction of 0.30 or less. Patients were randomly assigned in a 3:2 ratio to receive an implantable defibrillator (742 patients) or conventional medical therapy (490 patients). Invasive electrophysiological testing for risk stratification was not required. Death from any cause was the end point. RESULTS: The clinical characteristics at base line and the prevalence of medication use at the time of the last follow-up visit were similar in the two treatment groups. During an average follow-up of 20 months, the mortality rates were 19.8 percent in the conventional-therapy group and 14.2 percent in the defibrillator group. The hazard ratio for the risk of death from any cause in the defibrillator group as compared with the conventional-therapy group was 0.69 (95 percent confidence interval, 0.51 to 0.93; P=0.016). The effect of defibrillator therapy on survival was similar in subgroup analyses stratified according to age, sex, ejection fraction, New York Heart Association class, and the QRS interval. CONCLUSIONS: In patients with a prior myocardial infarction and advanced left ventricular dysfunction, prophylactic implantation of a defibrillator improves survival and should be considered as a recommended therapy. PMID- 11907287 TI - Amiodarone as compared with lidocaine for shock-resistant ventricular fibrillation. AB - BACKGROUND: Lidocaine has been the initial antiarrhythmic drug treatment recommended for patients with ventricular fibrillation that is resistant to conversion by defibrillator shocks. We performed a randomized trial comparing intravenous lidocaine with intravenous amiodarone as an adjunct to defibrillation in victims of out-of-hospital cardiac arrest. METHODS: Patients were enrolled if they had out-of-hospital ventricular fibrillation resistant to three shocks, intravenous epinephrine, and a further shock; or if they had recurrent ventricular fibrillation after initially successful defibrillation. They were randomly assigned in a double-blind manner to receive intravenous amiodarone plus lidocaine placebo or intravenous lidocaine plus amiodarone placebo. The primary end point was the proportion of patients who survived to be admitted to the hospital. RESULTS: In total, 347 patients (mean [+/-SD] age, 67+/-14 years) were enrolled. The mean interval between the time at which paramedics were dispatched to the scene of the cardiac arrest and the time of their arrival was 7+/-3 minutes, and the mean interval from dispatch to drug administration was 25+/-8 minutes. After treatment with amiodarone, 22.8 percent of 180 patients survived to hospital admission, as compared with 12.0 percent of 167 patients treated with lidocaine (P=0.009; odds ratio, 2.17; 95 percent confidence interval, 1.21 to 3.83). Among patients for whom the time from dispatch to the administration of the drug was equal to or less than the median time (24 minutes), 27.7 percent of those given amiodarone and 15.3 percent of those given lidocaine survived to hospital admission (P=0.05). CONCLUSIONS: As compared with lidocaine, amiodarone leads to substantially higher rates of survival to hospital admission in patients with shock-resistant out-of-hospital ventricular fibrillation. PMID- 11907288 TI - Fluconazole for the treatment of cutaneous leishmaniasis caused by Leishmania major. AB - BACKGROUND: Whereas certain oral antifungal azoles are well documented to have activity against leishmania, data on the efficacy of fluconazole for leishmaniasis are limited. We conducted a controlled trial in Saudi Arabia of fluconazole for the treatment of cutaneous leishmaniasis caused by Leishmania major. METHODS: This randomized, double-blind, placebo-controlled trial assessed the efficacy of oral fluconazole, in a dose of 200 mg daily for six weeks, in the treatment of parasitologically confirmed cutaneous leishmaniasis. The primary outcome measure was the time to the complete healing of all lesions. RESULTS: A total of 106 patients were assigned to receive fluconazole, and 103 patients were assigned to receive placebo. Follow-up data were available for 80 and 65 patients, respectively. At the three-month follow-up, healing of lesions was complete for 63 of the 80 patients in the fluconazole group (79 percent) and 22 of the 65 patients in the placebo group (34 percent; relative risk of complete healing, 2.33 [95 percent confidence interval, 1.63 to 3.33]). According to an intention-to-treat analysis, the rates of healing were 59 percent and 22 percent, respectively (relative risk, 2.76 [95 percent confidence interval, 1.84 to 4.12]). Sodium stibogluconate was offered to 11 patients in the fluconazole group who returned for follow-up (14 percent) and 33 of those in the placebo group (51 percent) in whom oral treatment was judged to have failed. According to a Kaplan Meier analysis, the time to healing was shorter for the fluconazole group (median, 8.5 weeks, as compared with 11.2 weeks in the placebo group; P<0.001 by the log-rank test). Side effects were mild and similar in both groups. CONCLUSIONS: A six-week course of oral fluconazole is a safe and useful treatment for cutaneous leishmaniasis caused by L. major. PMID- 11907289 TI - Bosentan therapy for pulmonary arterial hypertension. AB - BACKGROUND: Endothelin-1 is a potent vasoconstrictor and smooth-muscle mitogen. In a preliminary study, the orally administered dual endothelin-receptor antagonist bosentan improved exercise capacity and cardiopulmonary hemodynamics in patients with pulmonary arterial hypertension. The present trial investigated the effect of bosentan on exercise capacity in a larger number of patients and compared two doses. METHODS: In this double-blind, placebo-controlled study, we randomly assigned 213 patients with pulmonary arterial hypertension (primary or associated with connective-tissue disease) to receive placebo or to receive 62.5 mg of bosentan twice daily for 4 weeks followed by either of two doses of bosentan (125 or 250 mg twice daily) for a minimum of 12 weeks. The primary end point was the degree of change in exercise capacity. Secondary end points included the change in the Borg dyspnea index, the change in the World Health Organization (WHO) functional class, and the time to clinical worsening. RESULTS: At week 16, patients treated with bosentan had an improved six-minute walking distance; the mean difference between the placebo group and the combined bosentan groups was 44 m (95 percent confidence interval, 21 to 67; P<0.001). Bosentan also improved the Borg dyspnea index and WHO functional class and increased the time to clinical worsening. CONCLUSIONS: The endothelin-receptor antagonist bosentan is beneficial in patients with pulmonary arterial hypertension and is well tolerated at a dose of 125 mg twice daily. Endothelin-receptor antagonism with oral bosentan is an effective approach to therapy for pulmonary arterial hypertension. PMID- 11907290 TI - Images in clinical medicine. Fistula connecting the left main coronary artery with the right atrium in a marathon runner. PMID- 11907291 TI - A controlled trial of inpatient and outpatient geriatric evaluation and management. AB - BACKGROUND: Over the past 20 years, both inpatient units and outpatient clinics have developed programs for geriatric evaluation and management. However, the effects of these interventions on survival and functional status remain uncertain. METHODS: We conducted a randomized trial involving frail patients 65 years of age or older who were hospitalized at 11 Veterans Affairs medical centers. After their condition had been stabilized, patients were randomly assigned, according to a two-by-two factorial design, to receive either care in an inpatient geriatric unit or usual inpatient care, followed by either care at an outpatient geriatric clinic or usual outpatient care. The interventions involved teams that provided geriatric assessment and management according to Veterans Affairs standards and published guidelines. The primary outcomes were survival and health-related quality of life, measured with the use of the Medical Outcomes Study 36-Item Short-Form General Health Survey (SF-36), one year after randomization. Secondary outcomes were the ability to perform activities of daily living, physical performance, utilization of health services, and costs. RESULTS: A total of 1388 patients were enrolled and followed. Neither the inpatient nor the outpatient intervention had a significant effect on mortality (21 percent at one year overall), nor were there any synergistic effects between the two interventions. At discharge, patients assigned to the inpatient geriatric units had significantly greater improvements in the scores for four of the eight SF-36 subscales, activities of daily living, and physical performance than did those assigned to usual inpatient care. At one year, patients assigned to the outpatient geriatric clinics had better scores on the SF-36 mental health subscale, even after adjustment for the score at discharge, than those assigned to usual outpatient care. Total costs at one year were similar for the intervention and usual-care groups. CONCLUSIONS: In this controlled trial, care provided in inpatient geriatric units and outpatient geriatric clinics had no significant effects on survival. There were significant reductions in functional decline with inpatient geriatric evaluation and management and improvements in mental health with outpatient geriatric evaluation and management, with no increase in costs. PMID- 11907292 TI - Subtle acquired renal injury as a mechanism of salt-sensitive hypertension. PMID- 11907293 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 9-2002. An 80-year-old woman with sudden unilateral blindness. PMID- 11907294 TI - Expanding indications for implantable cardiac defibrillators. PMID- 11907295 TI - Treatment of primary pulmonary hypertension -- the next generation. PMID- 11907296 TI - The future of the global tobacco treaty negotiations. PMID- 11907297 TI - Prevention of venous thromboembolism with fondaparinux. PMID- 11907298 TI - Estrogen-replacement therapy after ischemic stroke. PMID- 11907299 TI - Recognition and management of anthrax. PMID- 11907300 TI - Cutaneous anthrax infection. PMID- 11907301 TI - Sudden death due to cardiac arrhythmias. PMID- 11907302 TI - Red pepper and functional dyspepsia. PMID- 11907304 TI - Optident prize 2000. AB - This paper describes the orthodontic treatment of two cases, which were awarded the 2000 Optident prize. PMID- 11907305 TI - Maxillo-nasal dysplasia, Binder's syndrome: review of the literature and case report. AB - A 12-year-old girl with maxillo-nasal dysplasia (Binder's syndrome), featuring maxillary hypoplasia and relative mandibular prognathism, presented with a Class III incisal relationship. Her treatment was managed orthodontically. The principal features of the syndrome and management of these cases is discussed. PMID- 11907306 TI - The craniofacial morphology of the parents of children with orofacial clefting: a systematic review of cephalometric studies. AB - OBJECTIVE: To systematically review the cephalometric studies investigating the craniofacial morphology of the parents of children with orofacial clefting (OFC). SEARCH STRATEGY: The search strategy was based on the keywords 'parent', cephalometry', and 'cleft', identifying 17 studies, of which 15 'case/control' studies met the inclusion criteria Statistically significant clinically relevant cephalometric variables from univariate statistical tests and multivariate results were collated and presented unweighted. RESULTS/CONCLUSIONS: The parental craniofacial complex in OFC is distinctive in comparison to the non-cleft population. However, there is insufficient consistency in study designs and results to accurately characterize the parents of children with OFC. Although the craniofacial morphology of the parents of children with CL(P) differs to the parents of children with CP, there is insufficient information to accurately localize these differences. PMID- 11907307 TI - A randomized clinical trial to compare three methods of orthodontic space closure. AB - AIM: To compare the rates of orthodontic space closure for: Active ligatures, polyurethane powerchain (Rocky Mountain Orthodontics, RMO Europe, Parc d'Innovation, Rue Geiler de Kaysersberg, 67400 Illkirch-Graffenstaden, Strasbourg, France) and nickel titanium springs. SAMPLE: Patients entering the space closure phase of fixed orthodontic treatment attending six orthodontic providers. Twelve patients received active ligatures (48 quadrants), 10 patients received powerchain (40 quadrants) and 11 patients, nickel-titanium springs (44 quadrants). METHOD: Patients were randomly allocated for treatment with active ligatures, powerchain or nickel titanium springs. Upper and lower study models were collected at the start of space closure (T(o)) and 4 months later (T(1)). We recorded whether the patient wore Class II or Class III elastics. Space present in all four quadrants was measured, by a calibrated examiner, using Vernier callipers at T(o) and T(1.) The rate of space closure, in millimetres per month (4 weeks) and a 4-monthly rate, was then calculated. Examiner reliability was assessed at least 2 weeks later. RESULTS: Mean rates of space closure were 0.35 mm/month for active ligatures, 0.58 mm/month for powerchain, and 0.81 mm/month for NiTi springs. No statistically significant differences were found between any methods with the exception of NiTi springs showing more rapid space closure than active ligatures (P < 0.05). There was no effect of inter-arch elastics on rate of space closure. CONCLUSIONS: NiTi springs gave the most rapid rate of space closure and may be considered the treatment of choice. However, powerchain provides a cheaper treatment option that is as effective. The use of inter-arch elastics does not appear to influence rate of space closure. PMID- 11907308 TI - Strength decay of orthodontic elastomeric ligatures. AB - AIM: To evaluate, over a 12-week period, the tensile strength (TS) and extension to TS of elastomeric ligatures (both clear and coloured) obtained from two companies. SETTING: Ex vivo study. METHOD: Seven replicates of five ligatures from two orthodontic companies were tested using an ex vivo assembly which simulated a clinical situation. RESULTS: The ligatures tested had a similar TS decay pattern, with the TS gradually decreasing over the duration of the study. The Unitek ligatures exhibited a greater TS than their respective Ormco ligatures. Generally, all of the ligatures tested experienced an increase in the extension to TS over time. The pre-stretching procedure used in this study did not produce any long-term detrimental effects on the TS and extension to TS of the ligatures tested. CONCLUSIONS: The results of this study confirm there are differences in TS, and extension to TS between clear and coloured elastomeric ligatures, and that some significant differences also exist between different brands of elastomeric ligatures. PMID- 11907309 TI - Halogen versus high-intensity light-curing of uncoated and pre-coated brackets: a shear bond strength study. AB - AIM: To evaluate the shear bond strengths of adhesive pre-coated brackets (APC) and conventional uncoated brackets (Victory) cured with two different light curing units: a conventional halogen light (Visilux 2) and a micro-xenon light (Aurys). SETTING: Ex vivo study MATERIALS AND METHODS: Sixty freshly extracted bovine permanent mandibular incisors were randomly assigned to one of four groups, each group consisting of 15 specimens. Two groups (one for each type of bracket) were exposed to the halogen light for 20 seconds and used as controls. The remaining two groups were cured with the micro-xenon light for 2 seconds. After 24 hours, all samples were tested in a shear mode on an Instron Machine. Analysis was by two-way ANOVA with Scheffe's test for comparisons, Kaplan-Meier survival estimates, and Cox model. The Chi-square (chi(2)) test was used to determine significant differences in the ARI scores. RESULTS: The mean shear bond strength of the uncoated brackets cured with Visilux 2 was significantly higher than those of all the other groups tested. Both groups cured with Visilux 2 produced significantly higher mean shear bond strengths than those of the corresponding groups cured with Aurys. No statistically significant differences were found between the two groups cured with Aurys. CONCLUSIONS: Compared to halogen light-curing, the micro-xenon light enables the clinician to reduce significantly the curing time of both APC and uncoated brackets, and although significantly lower, their shear bond strengths may be clinically acceptable. PMID- 11907312 TI - How to write a protocol. PMID- 11907310 TI - A clinical trial of light cure acrylic resin for orthodontic use. AB - OBJECTIVE: To ascertain whether or not the less porous surface associated with visible light cure appliances and the absence of free monomer had any measurable affect upon mucosal erythema, and to assess the durability of such appliances in a clinical context. DESIGN: A prospective randomized trial of visible light cure (Triad VLC) and autopolymerizing (Orthoresin) acrylic resin used as orthodontic base plate materials. SETTING: University Dental Hospital and School. SUBJECTS: Fifty subjects from a consecutively enrolled sample of 69 (19 drop outs) for removable appliance therapy (23 VLC, 27 AP). OUTCOME MEASURES: Erythema meter scores and appliance breakages. RESULTS: No statistical difference in mucosal erythema between the two materials was found. Fifty-two per cent of VLC appliances broke during a 6-month period, as opposed to 7 per cent of AP appliances. CONCLUSIONS: VLC appears to have no clinically beneficial effect on the oral mucosa compared with AP. VLC appliances are currently not sufficiently durable to make them a viable alternative to AP appliances. PMID- 11907313 TI - Orthodontics in China. PMID- 11907318 TI - Averageness as the ideal. PMID- 11907314 TI - An introduction to digital radiography in dentistry. AB - The aim of this article is to explain the basic principles of digital radiography, and to discuss the intra- and extra-oral imaging systems currently available. There are two main types of digital sensors available. One is based on charge coupled device technology and the other consists of phosphor storage plates. The advantages and disadvantages of each are highlighted with particular attention to orthodontics. PMID- 11907319 TI - Undergraduate curriculum in orthodontics. PMID- 11907320 TI - The influenza virus nucleoprotein: a multifunctional RNA-binding protein pivotal to virus replication. AB - All viruses with negative-sense RNA genomes encode a single-strand RNA-binding nucleoprotein (NP). The primary function of NP is to encapsidate the virus genome for the purposes of RNA transcription, replication and packaging. The purpose of this review is to illustrate using the influenza virus NP as a well-studied example that the molecule is much more than a structural RNA-binding protein, but also functions as a key adapter molecule between virus and host cell processes. It does so through the ability to interact with a wide variety of viral and cellular macromolecules, including RNA, itself, two subunits of the viral RNA dependent RNA polymerase and the viral matrix protein. NP also interacts with cellular polypeptides, including actin, components of the nuclear import and export apparatus and a nuclear RNA helicase. The evidence for the existence of each of these activities and their possible roles in transcription, replication and intracellular trafficking of the virus genome is considered. PMID- 11907322 TI - Differentiation of monocytes to macrophages induced by influenza virus-infected apoptotic cells. AB - The effect of the culture supernatant of influenza virus (IV)-infected apoptotic and non-apoptotic cells on the differentiation of monocytes to macrophages was investigated. IV infection induced apoptotic DNA fragmentation in cultured chorion cells but not in amnion cells prepared from human foetal membrane tissue. To examine the differentiation of monocytes to macrophages, an adhesion assay was employed using the human monocytic leukaemia THP-1 cell line. THP-1 cells became adherent to a substrate by incubation with the culture supernatant of IV-infected chorion cells, but not with that of amnion cells. The spreading THP-1 cells were morphologically characteristic of macrophages and they phagocytosed latex particles. RT-PCR analysis revealed that the expression of class A scavenger receptor mRNA was induced in THP-1 cells by incubation with the culture supernatant of IV-infected chorion cells. These results suggested that monocytic THP-1 cells were morphologically and functionally differentiated to macrophages by IV-infected apoptotic cells due to a soluble factor released from the apoptotic cells. PMID- 11907321 TI - Antigenic and genetic diversity among swine influenza A H1N1 and H1N2 viruses in Europe. AB - Three subtypes of influenza A viruses, H1N1, H1N2 and H3N2, co-evolve in pigs in Europe. H1N2 viruses isolated from pigs in France and Italy since 1997 were closely related to the H1N2 viruses which emerged in the UK in 1994. In particular, the close relationship of the neuraminidases (NAs) of these viruses to the NA of a previous UK H3N2 swine virus indicated that they had not acquired the NA from H3N2 swine viruses circulating in continental Europe. Moreover, antigenic and genetic heterogeneity among the H1N2 viruses appeared to be due in part to multiple introductions of viruses from the UK. On the other hand, comparisons of internal gene sequences indicated genetic exchange between the H1N2 viruses and co-circulating H1N1 and/or H3N2 subtypes. Most genes of the earlier (1997-1998) H1N2 isolates were more closely related to those of a contemporary French H1N1 isolate, whereas the genes of later (1999-2000) isolates, including the HAs of some H1N2 viruses, were closely related to those of a distinct H1N1 antigenic variant which emerged in France in 1999. In contrast, an H3N2 virus isolated in France in 1999 was closely related antigenically and genetically to contemporary human A/Sydney/5/97-like viruses. These studies reveal interesting parallels between genetic and antigenic drift of H1N1 viruses in pig and human populations, and provide further examples of the contribution of genetic reassortment to the antigenic and genetic diversity of swine influenza viruses and the importance of the complement of internal genes in the evolution of epizootic strains. PMID- 11907323 TI - Respiratory syncytial virus matrix protein associates with nucleocapsids in infected cells. AB - Little is known about the functions of the matrix (M) protein of respiratory syncytial virus (RSV). By analogy with other negative-strand RNA viruses, the M protein should inhibit the viral polymerase prior to packaging and facilitate virion assembly. In this study, localization of the RSV M protein in infected cells and its association with the RSV nucleocapsid complex was investigated. RSV infected cells were shown to contain characteristic cytoplasmic inclusions. Further analysis showed that these inclusions were localization sites of the M protein as well as the N, P, L and M2-1 proteins described previously. The M protein co-purified with viral ribonucleoproteins (RNPs) from RSV-infected cells. The transcriptase activity of purified RNPs was enhanced by treatment with antibodies to the M protein in a dose-dependent manner. These data suggest that the M protein is associated with RSV nucleocapsids and, like the matrix proteins of other negative-strand RNA viruses, can inhibit virus transcription. PMID- 11907324 TI - Hantavirus nucleocapsid protein interacts with the Fas-mediated apoptosis enhancer Daxx. AB - Hantaviruses cause two severe diseases, haemorrhagic fever with renal syndrome in Eurasia and hantavirus pulmonary syndrome in the Americas. To understand more about the molecular mechanisms that lead to these diseases, the associations of Puumala virus nucleocapsid protein (PUUV-N) with cellular proteins were studied by yeast two-hybrid screening. Daxx, known as an apoptosis enhancer, was identified from a HeLa cDNA library and its interaction with PUUV-N was confirmed by GST pull-down assay, co-immunoprecipitation and co-localization studies. Furthermore, domains of interaction were mapped to the carboxyl-terminal region of 142 amino acids in Daxx and the carboxyl-terminal 57 residues in PUUV-N, respectively. In pepscan assays, the binding sites of Daxx to PUUV-N were mapped further to two lysine-rich regions, of which one overlaps the sequence of the predicted nuclear localization signal of Daxx. These data suggest a direct link between host cell machinery and a hantavirus structural component. PMID- 11907325 TI - Identification of a cell surface 30 kDa protein as a candidate receptor for Hantaan virus. AB - Cellular receptors play an important role in virus pathogenesis. As a first step in virus infection, viruses attach to specific receptors on the surface of cells; Hantaan virus infects susceptible cells by attaching to a receptor located on the cell surface. To date, the identity of the Hantaan virus host cell receptor remains unknown. To determine the protein on the cell surface to which Hantaan virus binds, a virus overlay protein-binding assay was performed with radiolabelled virus. A 30 kDa (30K) protein was identified as a putative receptor for Hantaan virus. The specificity of virus interactions with this protein was demonstrated with a competition assay using unlabelled Hantaan virus and poliovirus. Unlabelled Hantaan virus inhibited the binding of radiolabelled Hantaan virus to this 30 kDa protein, whereas poliovirus did not. A polyclonal antibody against the 30K protein blocked binding of Hantaan virus to Vero-E6 cells and, consequently, virus infection. Blocking with the anti-30K antibody reduced virus infection of cells by 70%. These data strongly suggest that the 30 kDa surface protein is a putative receptor for Hantaan virus. PMID- 11907326 TI - Antigenic sites of foot-and-mouth disease virus (FMDV): an analysis of the specificities of anti-FMDV antibodies after vaccination of naturally susceptible host species. AB - Of the known neutralizing antigenic sites of foot-and-mouth disease virus (FMDV), site 1 or A, formed in part by the G-H loop of VP1, has historically been considered immunodominant because of evidence implicating its importance in the induction of a protective immune response. However, no systematic study has been done to determine the relative importance of the various specificities of antibodies against the known neutralizing antigenic sites of FMDV in the polyclonal immune response of a natural host after vaccination. In this report, we have adopted a monoclonal antibody-based competition ELISA and used antibodies specific to sites 1, 2 and 3 to provide some insight into this issue. Following vaccination of the three main target species, cattle, pigs and sheep, with an O1 serotype strain, results indicate that none of these three antigenic sites can be considered immunodominant in a polyclonal serum. Interestingly, pigs did not respond to epitopes on the carboxy terminus end of VP1 as efficiently as the ruminant species. In addition to the known sites, other as yet undefined sites might also be important in the induction of a protective immune response. Possible implications for the design of new vaccine strategies for foot-and-mouth disease are discussed. PMID- 11907327 TI - Multimerization reactions of coxsackievirus proteins 2B, 2C and 2BC: a mammalian two-hybrid analysis. AB - Recently, homomultimerization and heteromultimerization reactions of the poliovirus P2 region proteins were investigated using a yeast two-hybrid approach (Cuconati et al., Journal of Virology 72, 1297-1307, 1998). In this study, we investigated multimerization reactions of the 2B, 2C and 2BC proteins of the closely related coxsackie B3 virus (CBV3) using a mammalian two-hybrid system. This system allows the characterization of protein:protein interactions within a cellular environment that more closely mimics the native protein environment. Homomultimerization reactions were observed with the 2BC protein and, albeit weakly, with the 2B protein, but not with the 2C protein. To identify the determinants involved in the 2BC and 2B homomultimerization reactions, several mutants containing deletions or point mutations in the 2B region were tested. Disruption of the hydrophobic character of either the cationic amphipathic alpha helix or the second hydrophobic domain of the 2B protein disturbed both the 2BC:2BC and the 2B:2B homomultimerization reactions. Disruption of either the cationic or the amphipathic character of the alpha-helix or deletion of the N terminal 30 amino acids of the 2B protein, however, had no effect on the 2BC and 2B homomultimerization reactions. Heteromultimerization reactions were observed between proteins 2BC and 2B, and also between proteins 2BC and 2C, but not between the 2B and 2C proteins. The 2BC:2B and 2BC:2C heteromultimerization reactions were also mediated by hydrophobic determinants located in the amphipathic alpha-helix and the second hydrophobic domain. The nature of the interactions and their implications for the virus life-cycle are discussed. PMID- 11907328 TI - Nuclear localization of non-structural protein 1 and nucleocapsid protein of equine arteritis virus. AB - RNA synthesis (genome replication and subgenomic mRNA transcription) directed by equine arteritis virus (EAV; family Arteriviridae, order Nidovirales) occurs on modified cytoplasmic membranes to which most viral replicase subunits localize. Remarkably, a fraction of non-structural protein 1 (nsp1), a protein essential for transcription but dispensable for genome replication, is present in the host cell nucleus, in particular during the earlier stages of infection. Expression of GFP-tagged fusion proteins revealed that nsp1 is actively imported into the nucleus. Although the signals responsible for nsp1 transport could not be identified, our studies revealed that another EAV protein with a partially nuclear localization, the nucleocapsid (N) protein, utilizes the CRM1-mediated nuclear export pathway. Inactivation of this pathway with the drug leptomycin B resulted in the unexpected and immediate nuclear retention of all N protein molecules, thus revealing that the protein shuttles between cytoplasm and nucleus before playing its role in cytoplasmic virus assembly. PMID- 11907329 TI - Sustained G-->A hypermutation during reverse transcription of an entire human immunodeficiency virus type 1 strain Vau group O genome. AB - Two full-length human immunodeficiency virus type 1 O sequences are described, one of which was hypermutated in all regions of the genome. This indicates that the intracellular [dTTP]/[dCTP] bias conducive to G-->A hypermutation may be sustained throughout the synthesis of minus-strand DNA. In turn, this suggests the possibility of mutation of host sequences. PMID- 11907330 TI - Immunization with recombinant modified vaccinia virus Ankara can modify mucosal simian immunodeficiency virus infection and delay disease progression in macaques. AB - In the present study, the immunogenicity and protective efficacy of a recombinant vaccinia virus-based simian immunodeficiency virus (SIV) vaccine, given alone or in combination with a protein boost, were investigated. Cynomolgus macaques were immunized intramuscularly with modified vaccinia virus Ankara (MVA) expressing the SIVsm env and gag-pol genes (MVA-SIVsm) at 0 and 3 months (n=4), at 0, 3 and 8 months (n=4) or at 0 and 3 months followed by purified native SIVsm gp148 and recombinant SIVmac p27 in immunostimulatory complexes at 8 months (n=4). One month after the last immunization, the vaccinees, together with four naive control monkeys and four monkeys immunized with wild-type MVA, were challenged intrarectally with 10 MID50 SIVsm. At the time of challenge, antibody titres to SIV Env and lymphocyte proliferation responses to whole viral antigen were highest in vaccinees receiving MVA-SIVsm in combination with protein immunizations. Following rectal challenge, one of these vaccinees was completely protected. A prolonged survival time was observed in two of four monkeys in each of the groups immunized with MVA-SIVsm, in two monkeys given MVA-SIVsm followed by protein and in three of four monkeys given wild-type MVA, compared with naive controls. In conclusion, one monkey given the combined vaccine was protected completely against SIVsm infection. Furthermore, immunization with MVA-SIVsm, as well as wild-type MVA alone, seemed to delay disease progression after mucosal SIV infection in a proportion of the monkeys. PMID- 11907331 TI - Linkage on chromosome 10 of several murine retroviral integration loci associated with leukaemia. AB - Mml loci have been identified as provirus integration sites among a subset of monocytic tumours induced by murine leukaemia virus (MuLV) infection of BALB/c and DBA/2 mice. These myeloid leukaemias contain a retrovirus integrated on chromosome 10 in proximity to the c-myb locus; however, c-myb expression was not altered. Detailed physical mapping enabled placement of the retroviral integration sites approximately 25 kb (Mml1), approximately 51 kb (Mml2), and approximately 70 kb (Mml3) upstream of the c-myb locus. Furthermore, the Fti1 (fit-1) locus, a common integration site in feline leukaemia virus-induced T cell lymphomas, was mapped upstream of Mml3. Sequence analysis of Mml1, Mml2 and Mml3 loci (39.6, 16.4 and 5.9 kb, respectively) in conjunction with the BLAST (basic local alignment search tool) homology searches against the expressed sequence tag (EST) database and the use of gene/exon prediction programs revealed potential coding sequences that were not confirmed by Northern analysis or RT-PCR. The sequences between c-myb and Fti1, which were shown to include two potential scaffold/matrix attachment regions (S/MARs), are most likely regulatory in nature. An extended search for transcribed sequences far upstream of Mml3 revealed five genes, four of which were expressed in multiple tissues in mice. These genes could not be linked to tumour formation by the virus but their homologous sequences were found on human chromosome 6, thus allowing extension of the syntenic region on mouse chromosome 10 to approximately 250 kb. PMID- 11907332 TI - Complementation of a p300/CBP defective-binding mutant of adenovirus E1a by human papillomavirus E6 proteins. AB - Previous studies have shown that the human papillomavirus type 16 (HPV-16) E6 protein binds to p300/CBP and abrogates its transcriptional co-activator function. However, there is little information on the biological consequences of this interaction and discrepancy as to whether the interaction is high-risk E6 specific or not. We performed a series of studies to compare the interactions of HPV-18 and HPV-11 E6 with p300, and showed that both high- and low- risk E6 proteins bind p300. In addition, using a transformation-deficient mutant of adenovirus E1a, which cannot interact with p300, we demonstrated that HPV-16, HPV 18 and, to a lesser extent, HPV-11 E6, can complement this mutant in cell transformation assays. In contrast, a mutant of HPV-16 E6 which does not bind p300 failed to rescue the E1a mutant. These results suggest that the E6-p300 interaction may be important for the ability of HPV E6 to contribute towards cell transformation. PMID- 11907333 TI - Analysis of the complete genome of subgroup A' hepatitis B virus isolates from South Africa. AB - A phylogenetic analysis is presented of six complete and seven pre-S1/S2/S gene sequences of hepatitis B virus (HBV) isolates from South Africa. Five of the full length sequences and all of the pre-S2/S sequences have been previously reported. Four of the six complete genomes and three of the five incomplete sequences clustered with subgroup A', a unique segment of genotype A of HBV previously identified in 60% of South African isolates using analysis of the pre-S2/S region alone. This separation was also evident when the polymerase open reading frame was analysed, but not on analysis of either the X or pre-core/core genes. Amino acids were identified in the pre-S1 and polymerase regions specific to subgroup A'. In common with genotype D, 10 of 11 genotype A South African isolates had an 11 amino acid deletion in the amino end of the pre-S1 region. This deletion is also found in hepadnaviruses from non-human primates. PMID- 11907334 TI - Viral gene expression during acute simian varicella virus infection. AB - Simian varicella virus (SVV) causes a natural varicella-like disease in nonhuman primates. Outbreaks of simian varicella occur sporadically in primate facilities. Simian varicella is used as a model for investigation of varicella-zoster virus (VZV) pathogenesis and latency. In this study, SVV gene expression and histopathology were analysed in tissues of acutely infected vervet monkeys. RT PCR analysis demonstrated expression of specific SVV immediate early, early and late genes in the skin, lung, liver and ganglia tissues of acutely infected monkeys. Viral antigen expression and histopathology, including necrosis and inflammation, were detected in the skin, lungs, liver and spleen of infected monkeys by immunohistochemical analysis. Viral antigen expression, but little or no histopathology, was evident in the neural ganglia, the eventual site of viral latency. The study provides a foundation for further investigation on the role of viral genes in varicella pathogenesis and latency. PMID- 11907336 TI - The sequence of camelpox virus shows it is most closely related to variola virus, the cause of smallpox. AB - Camelpox virus (CMPV) and variola virus (VAR) are orthopoxviruses (OPVs) that share several biological features and cause high mortality and morbidity in their single host species. The sequence of a virulent CMPV strain was determined; it is 202182 bp long, with inverted terminal repeats (ITRs) of 6045 bp and has 206 predicted open reading frames (ORFs). As for other poxviruses, the genes are tightly packed with little non-coding sequence. Most genes within 25 kb of each terminus are transcribed outwards towards the terminus, whereas genes within the centre of the genome are transcribed from either DNA strand. The central region of the genome contains genes that are highly conserved in other OPVs and 87 of these are conserved in all sequenced chordopoxviruses. In contrast, genes towards either terminus are more variable and encode proteins involved in host range, virulence or immunomodulation. In some cases, these are broken versions of genes found in other OPVs. The relationship of CMPV to other OPVs was analysed by comparisons of DNA and predicted protein sequences, repeats within the ITRs and arrangement of ORFs within the terminal regions. Each comparison gave the same conclusion: CMPV is the closest known virus to variola virus, the cause of smallpox. PMID- 11907335 TI - Human herpesvirus-6 rep/U94 gene product has single-stranded DNA-binding activity. AB - The characterization is reported of the human herpesvirus-6B (HHV-6B) rep/U94 gene, which is a homologue of the adeno-associated virus type 2 rep. In this study, a monoclonal antibody was produced against HHV-6B REP (anti-REP mAb). Immunofluorescence staining using the anti-REP mAb showed that REP was localized to the nucleus in HHV-6-infected MT4 cells. It was first detected at 24 h post infection (p.i.) and accumulated to higher levels by 72 h p.i. REP may be expressed only at very low levels in HHV-6-infected cells: even when the late protein glycoprotein H was detected in nearly 90% of HHV-6-infected cells, REP was detected in only a small percentage of them. Western blot analysis showed that the anti-REP mAb recognized a 56-kDa polypeptide in HHV-6B-infected MT4 cells. Furthermore, the REP protein was shown to bind single-stranded DNA. PMID- 11907337 TI - The vaccinia virus B9R protein is a 6 kDa intracellular protein that is non essential for virus replication and virulence. AB - Vaccinia virus (VV) strain Western Reserve gene B9R is shown to encode an intracellular 6 kDa protein that is expressed late during the infectious cycle. In vitro transcription and translation produced two polypeptides in the presence of microsomal membranes, but only the larger protein in the absence of membranes. The smaller protein sedimented with microsomes during centrifugation, suggesting it was inserted into the lipid membrane or into the microsomal lumen via the N terminal hydrophobic signal sequence that was subsequently cleaved proteolytically. A VV mutant lacking B9R was constructed and found to replicate normally in cell culture and two in vivo models. PMID- 11907338 TI - Symptom induction by Cowpea chlorotic mottle virus on Vigna unguiculata is determined by amino acid residue 151 in the coat protein. AB - The type strain of Cowpea chlorotic mottle virus (CCMV-T) produces a bright chlorosis in cowpea (Vigna unguiculata cv. California Blackeye). The attenuated variant (CCMV-M) induces mild green mottle symptoms that were previously mapped to RNA 3. Restriction fragment exchanges between RNA 3 cDNA clones of CCMV-T and CCMV-M that generate infectious transcripts and site-directed mutagenesis indicated that the codon encoding amino acid residue 151 of the coat protein determines the symptom phenotypes of CCMV-T and CCMV-M. Amino acid 151 is within an alpha-helical structure required for calcium ion binding and virus particle stability. No differences in virion stability or accumulation were detected between CCMV-T and CCMV-M. Mutational analysis suggested that the amino acid at position 151 and not the nucleotide sequence induce the symptom phenotype. Thus, it is likely that subtle influences by amino acid residue 151 in coat protein host interactions result in chlorotic and mild green mottle symptoms. PMID- 11907339 TI - Characterization of plant proteins that interact with cowpea mosaic virus '60K' protein in the yeast two-hybrid system. AB - Cowpea mosaic virus (CPMV) replication occurs in close association with small membranous vesicles in the host cell. The CPMV RNA1-encoded 60 kDa nucleotide binding protein ('60K') plays a role in the formation of these vesicles. In this study, five cellular proteins were identified that interacted with different domains of 60K using a yeast two-hybrid search of an Arabidopsis cDNA library. Two of these host proteins (termed VAP27-1 and VAP27-2), with high homology to the VAP33 family of SNARE-like proteins from animals, interacted specifically with the C-terminal domain of 60K and upon transient expression colocalized with 60K in CPMV-infected cowpea protoplasts. eEF1-beta, picked up using the central domain of 60K, was also found to colocalize with 60K. The possible role of these host proteins in the viral replicative cycle is discussed. PMID- 11907341 TI - Adaptation from whitefly to leafhopper transmission of an autonomously replicating nanovirus-like DNA component associated with ageratum yellow vein disease. AB - Ageratum yellow vein disease is caused by the whitefly-transmitted monopartite begomovirus Ageratum yellow vein virus and a DNA beta satellite component. Naturally occurring symptomatic plants also contain an autonomously replicating nanovirus-like DNA 1 component that relies on the begomovirus and DNA beta for systemic spread and whitefly transmission but is not required for maintenance of the disease. Here, we show that systemic movement of DNA 1 occurs in Nicotiana benthamiana when co-inoculated with the bipartite begomovirus Tomato golden mosaic virus and the curtovirus Beet curly top virus (BCTV), but not with the mastrevirus Bean yellow dwarf virus. BCTV also mediates the systemic movement of DNA 1 in sugar beet, and the nanovirus-like component is transmitted between plants by the BCTV leafhopper vector Circulifer tenellus. We also describe a second nanovirus-like component, referred to as DNA 2, that has only 47% nucleotide sequence identity with DNA 1. The diversity and adaptation of nanovirus components are discussed. PMID- 11907340 TI - Phylogenetic relationships, strain diversity and biogeography of tritimoviruses. AB - North American and Eurasian isolates of Wheat streak mosaic virus (WSMV; genus Tritimovirus) and Oat necrotic mottle virus (ONMV; genus Rymovirus) were examined. Nine WSMV isolates differentially infected oat, barley, inbred maize line SDp2 and sorghum line KS56. The WSMV isolates clustered into groups based on phylogenetic analyses of the capsid protein (CP) cistron and flanking regions. WSMV isolates from the United States (US) and Turkey were closely related, suggesting recent movement between continents. Although more divergent, WSMV from Iran (WSMV-I) also shared a most recent common ancestor with the US and Turkish isolates. Another group of WSMV isolates from central Europe and Russia may represent a distinct Eurasian population. Complete genome sequences of WSMV from the Czech Republic (WSMV-CZ) and Turkey (WSMV-TK1) were determined and comparisons based on complete sequences yielded relationships similar to those based on partial sequences. ONMV-Pp recovered from blue grass (Poa pratensis L.) in Germany displayed the same narrow host range as ONMV-Type from Canada. Western blots revealed a heterologous relationship among CP of WSMV and ONMV. Phylogenetic analyses of the capsid protein cistron and flanking genomic regions indicated that WSMV and ONMV are related species sharing 74.2-76.2% (nucleotide) and 79.2-81.0% (amino acid) identity. Thus, ONMV should be removed from the genus Rymovirus and designated a definitive member of the genus Tritimovirus. Phylogenetic analyses further suggest that Sugarcane streak mosaic virus is not a tritimovirus, and may represent a new genus within the family Potyviridae. PMID- 11907343 TI - Gill-associated nidovirus of Penaeus monodon prawns transcribes 3'-coterminal subgenomic mRNAs that do not possess 5'-leader sequences. AB - Sequence analysis of the approximately 20 kb 5'-terminal portion of the ssRNA genome of gill-associated virus (GAV) of Penaeus monodon prawns has previously established that it contains an ORF1a-1b replicase gene equivalent to those of the coronavirus and arterivirus members of the order Nidovirales. Sequence analysis of the remaining approximately 6.2 kb of the GAV genome downstream of ORF1a-1b to a 3'-poly(A) tail has identified two highly conserved intergenic sequences in which 29/32 nucleotides are conserved. Northern hybridization using probes to the four putative GAV ORFs and either total or poly(A)-selected RNA identified two 3'-coterminal subgenomic (sg) mRNAs of approximately 6 kb and approximately 5.5 kb. Primer extension and 5'-RACE analyses showed that the sgmRNAs initiate at the same 5'-AC positions in the central region of the two conserved intergenic sequences. Neither method provided any evidence that the GAV sgmRNAs are fused to genomic 5'-leader RNA sequences as is the case with vertebrate coronaviruses and arteriviruses. Intracellular double-stranded (ds)RNAs equivalent in size to the 26.2 kb genomic RNA and two sgRNAs were also identified by RNase/DNase digestion of total RNA from GAV-infected prawn tissue. The identification of only two sgmRNAs that initiate at the same position in conserved intergenic sequences and the absence of 5'-genomic leader sequences fused to these sgmRNAs confirms that GAV has few genes and suggests that it utilizes a transcription mechanism possibly similar to the vertebrate toroviruses but distinct from coronaviruses and arteriviruses. PMID- 11907342 TI - Shrimp Taura syndrome virus: genomic characterization and similarity with members of the genus Cricket paralysis-like viruses. AB - The single-stranded genomic RNA of Taura syndrome virus (TSV) is 10205 nucleotides in length, excluding the 3' poly(A) tail, and contains two large open reading frames (ORFs) that are separated by an intergenic region of 207 nucleotides. The ORFs are flanked by a 377 nucleotide 5' untranslated region (UTR) and a 226 nucleotide 3' UTR followed by a poly(A) tail. The predicted amino acid sequence of ORF1 revealed sequence motifs characteristic of a helicase, a protease and an RNA-dependent RNA polymerase, similar to the non-structural proteins of several plant and animal RNA viruses. In addition, a short amino acid sequence located in the N-terminal region of ORF1 presented a significant similarity with a baculovirus IAP repeat (BIR) domain of inhibitor of apoptosis proteins from double-stranded DNA viruses and from animals. The presence of this BIR-like sequence is the first reported in a single-stranded RNA virus, but its function is unknown. The N-terminal amino acid sequence of three TSV capsid proteins (55, 40 and 24 kDa) were mapped in ORF2, which is not in the same reading frame as ORF1 and possesses an AUG codon upstream of the structural genes. However, the intergenic region shows nucleotide sequence similarity with those of the genus Cricket paralysis-like viruses, suggesting a similar non-AUG mediated translation mechanism. The structure of the TSV genome [5' UTR-non structural proteins-intergenic UTR-structural proteins-3' UTR-poly(A) tail] is similar to those of small insect-infecting RNA viruses, which were recently regrouped into a new virus genus, Cricket paralysis-like viruses. PMID- 11907344 TI - Transcriptional mapping of two genes encoding baculovirus envelope-associated proteins. AB - Genes encoding two representatives of the LD130 family of baculovirus envelope associated proteins were transcriptionally mapped. These included ld130, which encodes a low pH-induced envelope fusion protein of the Lymantria dispar multinucleocapsid nucleopolyhedrovirus, and op21, which is related to ld130 but is encoded by Orgyia pseudotsugata MNPV and appears to lack an envelope fusion activity. The size and temporal expression of mRNA of both genes were examined by Northern blot analysis of RNA extracted from infected cells at selected timepoints. In addition, 5' rapid amplification of cDNA ends (RACE) in combination with DNA sequence analysis was used to map the start sites of mRNA. Ld130 predominantly utilized its early promoter at 24 h post-infection but by 72 h post-infection ld130 expression was almost exclusively from its late promoter. In contrast, op21 was expressed predominantly from its early promoter throughout the timecourse, even though a consensus late promoter sequence was present within 100 bp of the translation start codon. A significant fraction of late transcripts that mapped to op21 were spliced transcripts originating in the op18 gene region. The 3' termini of the transcripts were also mapped using 3' RACE. PMID- 11907345 TI - Phenotypic and genotypic analysis of Helicoverpa armigera nucleopolyhedrovirus serially passaged in cell culture. AB - Rapid accumulation of few polyhedra (FP) mutants was detected during serial passaging of Helicoverpa armigera nucleopolyhedrovirus (HaSNPV) in cell culture. 100% FP infected cells were observed by passage 6. The specific yield decreased from 178 polyhedra per cell at passage 2 to two polyhedra per cell at passage 6. The polyhedra at passage 6 were not biologically active, with a 28-fold reduction in potency compared to passage 3. Electron microscopy studies revealed that very few polyhedra were produced in an FP infected cell (<10 polyhedra per section) and in most cases these polyhedra contained no virions. A specific failure in the intranuclear nucleocapsid envelopment process in the FP infected cells, leading to the accumulation of naked nucleocapsids, was observed. Genomic restriction endonuclease digestion profiles of budded virus DNA from all passages did not indicate any large DNA insertions or deletions that are often associated with such FP phenotypes for the extensively studied Autographa californica nucleopolyhedrovirus and Galleria mellonella nucleopolyhedrovirus. Within an HaSNPV 25K FP gene homologue, a single base-pair insertion (an adenine residue) within a region of repetitive sequences (seven adenine residues) was identified in one plaque-purified HaSNPV FP mutant. Furthermore, the sequences obtained from individual clones of the 25K FP gene PCR products of a late passage revealed point mutations or single base-pair insertions occurring throughout the gene. The mechanism of FP mutation in HaSNPV is likely similar to that seen for Lymantria dispar nucleopolyhedrovirus, involving point mutations or small insertions/deletions of the 25K FP gene. PMID- 11907346 TI - Whole genome analysis of the Epiphyas postvittana nucleopolyhedrovirus. AB - The nucleotide sequence of the Epiphyas postvittana nucleopolyhedrovirus (EppoMNPV) genome has been determined and analysed. The circular dsDNA genome contains 118584 bp, making it the smallest group I NPV sequenced to date. The genome has a G+C content of 40.7% and encodes 136 predicted open reading frames (ORFs), five homologous repeat regions and one unique repeat region. Of the genome, 92.9% encodes predicted ORFs and 2.2% is in repeat regions; the remaining 4.9% of the genome comprises nonrepeat intergenic regions. EppoMNPV encodes homologues of 126 Orgyia pseudotsugata MNPV (OpMNPV) ORFs and 120 Autographa californica MNPV ORFs, with average identities of 64.7 and 53.5%, respectively. Between the four sequenced group I NPVs, 117 ORFs are conserved, whereas 86 ORFs are conserved between all fully sequenced NPVs. A total of 62 ORFs is present in all baculoviruses sequenced to date, with EppoMNPV lacking a homologue of the superoxide dismutase (sod) gene, which has been found in all other fully sequenced baculoviruses. Whole genome phylogenetic analyses of the ten fully sequenced baculoviruses using the sequences of the 62 shared genes, gene content and gene order data sets confirmed that EppoMNPV clusters tightly with OpMNPV in the group I NPVs. The main variation between EppoMNPV and OpMNPV occurs where extra clusters of genes are present in OpMNPV, with sod occurring in one such cluster. EppoMNPV encodes one truncated baculovirus repeated ORF (bro) gene. The only repeated ORFs are the four iap genes. Eight, randomly distributed, unique ORFs were identified on EppoMNPV, none of which show any significant homology to genes in GenBank. PMID- 11907347 TI - Eugene Pasquale DiMagno, M.D. PMID- 11907348 TI - The acid achalasia association. PMID- 11907349 TI - Intestinal permeation and gastrointestinal disease. AB - The gastrointestinal tract constitutes one of the largest sites of exposure to the outside environment. The function of the gastrointestinal tract in monitoring and sealing the host interior from intruders is called the gut barrier. A variety of specific and nonspecific mechanisms are in operation to establish the host barrier; these include luminal mechanisms and digestive enzymes, the epithelial cells together with tight junctions in between them, and the gut immune system. Disruptions in the gut barrier follow injury from various causes including nonsteroidal anti-inflammatory drugs and oxidant stress, and involve mechanisms such as adenosine triphosphate depletion and damage to epithelial cell cytoskeletons that regulate tight junctions. Ample evidence links gut barrier dysfunction to multiorgan system failure in sepsis and immune dysregulation. Additionally, contribution of gut barrier dysfunction to gastrointestinal disease is an evolving concept and is the focus of this review. An overview of the evidence for the role of gut barrier dysfunction in disorders such as Crohn's disease, celiac disease, food allergy, acute pancreatitis, non-alcoholic fatty liver disease, and alcoholic liver disease is provided, together with critical insight into the implications of this evidence as a primary disease mechanism. PMID- 11907350 TI - Update in medical therapy of ulcerative colitis: a practical approach. AB - The bewildering array of medications in the therapy of inflammatory bowel disease (IBD) often confounds the clinician in the choice of specific agents regarding the balance between safety and efficacy. This review surveys and evaluates currently available IBD therapies as well as those used in clinical trials of ulcerative colitis. The primary purpose is to provide the busy clinician with a practical guide to the use of established and newly emerging medical therapies of IBD. PMID- 11907351 TI - Difference in prognostic value between sialyl Lewis(a) and sialyl Lewis(x) antigen levels in the preoperative serum of gastric cancer patients. AB - Sialyl Lewis(a) (CA19-9) and sialyl Lewis(x) antigens (SLX) may play a role in tumor metastasis by serving as functional ligands in the cell adhesion system. The authors examined preoperative serum levels of CA19-9 and SLX in 218 patients who underwent resection for gastric cancer to determine their prognostic value. The patients were divided into two groups, termed the low and high antigen groups, based on a value selected as a diagnostic cutoff. Correlation between the antigen serum levels, various established clinicopathologic factors, and prognosis were studied by univariate and multivariate analysis. The disease specific interval for high CA19-9 and SLX groups was significantly shorter than that of their respective low groups (p = 0.0024 and p < 0.0001, respectively). Patients with stage III/IV tumors who had high serum SLX levels had shorter disease-specific intervals than those with low serum levels (p = 0.0017). A Cox's regression analysis revealed a high serum SLX level as an independent factor for worse outcome. In addition, logistic regression analysis revealed that a high serum SLX level was an independent predictor for liver metastasis. In conclusion, an elevated preoperative serum SLX level was a predictor for poor outcome after resection for gastric cancer, whereas CA19-9 was not. PMID- 11907352 TI - Decreased production of interleukin-12 and type 2 immune responses are marked in cachectic patients with colorectal and gastric cancer. AB - Balance of the two types of T helper cells is one of the most important factors for regulation of the immune system. This study examines the production of interleukin (IL)-4, -6, -10, -12, and interferon-gamma by peripheral blood mononuclear cells stimulated with phytohemagglutinin or Staphylococcus aureus. Sixty-one patients, including 25 with gastric and 39 with colorectal cancer, and 39 normal volunteers were entered. The production of IL-12 decreased significantly with advancing disease and was lowest in the patients with distant metastases and cachexia. Compared with normal donors, the production of interferon-gamma decreased in all categories of patients, with no difference among patient groups. Levels of Th2 cytokines, such as IL-4, IL-6, and IL-10, also showed no difference among patient groups. However, production of all these cytokines had increased by 2.5 months after sequential testing in the same cachectic patients. The authors' findings indicate that the induction of Th1 cells seems to be suppressed at a relatively early stage of disease, whereas that of Th2 cells seems to increase in the terminal stage. PMID- 11907353 TI - Outcomes of endoscopy in patients with iron deficiency anemia after Billroth II partial gastrectomy. AB - GOALS: To determine the frequency of gastrointestinal lesions detected by upper endoscopy and colonoscopy in patients who developed iron deficiency anemia after Billroth II surgery. STUDY: The authors reviewed the medical records of 116 consecutive patients with a Billroth II partial gastrectomy and 232 age- and gender-matched controls without gastric surgery who were referred for endoscopy to evaluate iron deficiency anemia over a 5-year period. RESULTS: Clinically important lesions were detected in 22.4% of the patients with gastric surgery and in 59.5% of those with intact stomachs (p < 0.001). In the gastric surgery group, clinically important lesions were found more often in the upper gastrointestinal tract than in the colon (19.0% vs. 3.4%, p < 0.001). In the nonsurgical group, the diagnostic yields of upper endoscopy and colonoscopy were not significantly different (38.4% vs. 32.8%, p = 0.24). Synchronous lesions in the upper and lower gastrointestinal tract were significantly less common in the group of patients with gastric surgery compared with those without gastric surgery (0.0% vs. 11.6%, p < 0.001). Small bowel biopsies and small bowel follow-through did not identify any additional lesions. In the gastric surgery group, multivariate analysis identified abdominal symptoms (OR = 11.2, 95% CI 3.2-39.2, p < 0.001), a positive result on fecal occult blood testing (OR = 6.4, 95% CI 2.0-20.3, p = 0.002), and Billroth II surgery at least 10 years before evaluation (OR = 5.4, 95% CI 1.7 16.7, p = 0.004) as independent predictors of identifying a clinically important lesion by endoscopy. CONCLUSIONS: Upper endoscopy had a significantly higher diagnostic yield than colonoscopy in patients who developed iron deficiency anemia after Billroth II surgery. Prospective studies are necessary to determine the role and cost-effectiveness of colonoscopy in the evaluation of iron deficiency anemia in this patient population. PMID- 11907354 TI - Intestinal protein loss in acute and persistent diarrhea of early childhood. AB - GOALS: To determine fecal protein loss in children with acute and persistent diarrhea. BACKGROUND: In children with diarrhea, ongoing losses of endogenous proteins have been suggested as contributing to impairment of nutritional and immunologic status. However, there is a paucity of information and inconclusive data in the literature. STUDY: Fecal protein loss was assessed prospectively in children (<3 years of age) with acute diarrhea (<7 days' duration) or persistent diarrhea (>14 days) and in controls using alpha-1-antitrypsin determination; fecal protein loss then was correlated with age, duration of diarrhea, nutritional status, plasma proteins, and stool pathogens. RESULTS: Children with acute diarrhea (n = 43) and those with persistent diarrhea (n = 41) had significantly higher fecal alpha-1-antitrypsin levels compared with controls (n = 14) (2.26 +/- 1.71 and 2.25 +/- 1.51, respectively, vs. 1.02 +/- 0.73 mg/g stools; p = 0.002). However, there was no significant decrease of plasma albumin, globulin, or immunoglobulins. Fecal protein loss did not differ significantly among stool pathogens (bacterial, viral, and parasitic) and demonstrated no significant correlation with age, duration of diarrhea, or nutritional status (mild malnutrition). CONCLUSIONS: Enhanced fecal protein loss was observed in more than 50% of children with acute and persistent diarrhea caused by various pathogens. This did not correlate with age, duration of diarrhea, or nutritional status and did not result in significant decrease of plasma proteins or immunoglobulins. This protein-losing enteropathy does not appear to have a causal role in perpetuation of diarrheal episodes in children with mild malnutrition. PMID- 11907355 TI - Celiac disease in Brazilian adults. AB - Forty-eight adult patients with celiac disease between 15 and 68 years of age (mean, 41 years) were studied. Sixty-seven percent were female and 33% were male patients. Most of the patients were white (98%). The main clinical features were diarrhea (90%), weight loss (70%), and abdominal pain (56%). On physical examination, the main findings were pallor (40%), aphthous stomatitis (31%), and arthralgia (23%). Associated disorders included diabetes mellitus type I, osteoporosis, and atopy (6% each); dermatitis herpetiformis and depression (4% each); and hypothyroidism, hyperthyroidism, duodenal carcinoma, and Gilbert syndrome (2% each). The histologic results according to Marsh criteria (modified by Rostami) are as follows: type I, 10%; type II, 21%; type IIIa, 19%; type IIIb, 17%; and type IIIc, 33%. The sensitivity and specificity for the antiendomysium antibody-immunoglobulin A test were 92% and 100%, respectively, when considering the whole group of patients; however, the sensitivity (but not the specificity) decreased to 86% when taking into account only the group of patients with mild histologic alterations (Marsh type I, II, and IIIa). CONCLUSION: In general, the authors' results are similar to those described in developing countries, indicating that celiac disease might have the same spectrum of presentation regardless of the region studied. PMID- 11907356 TI - Clinical diagnosis of primary epiploic appendagitis: differentiation from acute diverticulitis. AB - BACKGROUND: Primary epiploic appendagitis (PEA) is an uncommon cause of abdominal pain that occurs either from appendageal torsion or spontaneous thrombosis of an appendageal draining vein. Primary epiploic appendagitis is frequently misdiagnosed as either appendicitis or diverticulitis, depending on its location. STUDY: Clinical and radiologic characteristics of 8 patients with PEA were retrospectively reviewed and compared with 18 patients with acute diverticulitis. RESULTS: Patients with PEA presented with lower abdominal pain of recent onset that was localized to the left (seven cases) and right (one case) lower quadrants. Well-localized tenderness without peritoneal irritation sign was usually the only physical finding. Blood tests were not significant. In acute diverticulitis, the pain was more evenly distributed throughout the lower abdomen and findings like nausea, fever, and leukocytosis were more frequently associated than in PEA. Computed tomography findings, such as pedunculated oval fatty mass with streaky densities connected to the serosal surface of the adjacent colon, can lead to the diagnosis of PEA. Symptoms of PEA were resolved within 1 week (mean, 4.7 days) without surgery. CONCLUSIONS: When patients with very localized lower abdominal pain and tenderness do not have associated symptoms or laboratory abnormalities, a high index of suspicion for PEA and early radiologic examinations are required. PMID- 11907357 TI - Barrett's esophagus and achalasia. AB - Two unusual cases of achalasia with endoscopic and histologic documentation of Barrett's esophagus are presented. One patient had Barrett's esophagus at the time of initial endoscopy for achalasia, before any treatment. The other patient developed specialized columnar epithelia in the esophagus after treatment with pneumatic dilation. Each patient had evidence of low-grade dysplasia. Including these two patients, 30 cases of Barrett's esophagus in patients with achalasia have been reported in the literature. In 73% (22 of 30) of the cases, Barrett's esophagus was detected after esophagomyotomy. In 20% (6 of 30) of the cases of achalasia and Barrett's esophagus, adenocarcinoma developed. The current two cases are unusual because Barrett's esophagus in achalasia generally develops from gastroesophageal reflux after esophagomyotomy. No other patients have been reported to develop Barrett's esophagus after pneumatic dilation alone. Patients with achalasia and Barrett's esophagus may be at a particularly high risk for developing dysplasia and adenocarcinoma. PMID- 11907358 TI - Gastrointestinal pneumatosis after docetaxel chemotherapy. AB - Breast cancer is one of the most frequent neoplasms in women. New drugs, including taxanes, have improved survival in patients with metastatic disease. Quality of life and efficacy are important goals during treatment of these women. Herein, we report a 51 year-old woman with metastatic breast cancer who developed gastrointestinal pneumatosis (GIP) after the first cycle of treatment, which consisted of docetaxel and pamidronate. The symptoms disappeared after 7 days with supportive management, nasogastric intubation, parenteral fluids, and wide spectrum antibiotics. Thereafter, weekly fractionated chemotherapy with an initial 50% dose reduction was administered. Because of adequate tolerance, the dose was increased by 25% after the second cycle, and full-dose docetaxel was administered after the third cycle. After 6 months of follow-up, the patient remained under treatment, with an Eastern Cooperative Oncology Group performance status of 1. Gastrointestinal pneumatosis has been reported in association with chemotherapy. In most patients, it is reported to be associated with neutropenia, which was not present in this patient. PMID- 11907359 TI - Mitochondrial neurogastrointestinal encephalomyopathy: diagnostic features of two patients. AB - Mitochondrial neurogastrointestinal encephalomyopathy is a rare, multisystem disorder characterized by gastrointestinal dysmotility, ptosis, neurologic findings (e.g., peripheral neuropathy), leukoencephalopathy, and thin body habitus. Gastrointestinal motility studies and skeletal muscle biopsy are recommended diagnostic tools. We report two patients that highlight the diagnostic characteristics of this rare entity. PMID- 11907360 TI - Turcot syndrome in an elderly adult. AB - A 67-year-old woman presented with hematochezia and an episode of transient expressive dysphasia. She was found to have multiple colonic polyps with adenocarcinomatous changes. Computed tomography brain scan and computed tomography-guided biopsy revealed a left frontoparietal glioblastoma multiforme. This case illustrates the rare presentation of Turcot syndrome-a hereditary colorectal polyposis syndrome-in an older adult. PMID- 11907361 TI - Rofecoxib: a possible cause of acute colitis. AB - Nonsteroidal antiinflammatory agents may cause relevant small bowel and colonic side effects, apart from gastroduodenal lesions. The synthesis of selective cyclooxygenase-2 (COX-2) inhibitors has been an important breakthrough in antiinflammatory therapy by decreasing the incidence of upper gastrointestinal lesions. However, there is little information available concerning their effects on gut mucosa distally to the duodenum. A case history is described of a 52-year old woman with a temporary colostomy after resection of a sigmoid tumor and who presented with bloody diarrhea 5 days after beginning therapy with rofecoxib. The hemorrhage had its origin in the transverse colon, and the endoscopic appearance was that of actively bleeding acute hemorrhagic colitis. No other colonic lesions were detected, nor was there any evidence of related infection, bleeding diathesis, or other systemic diseases. On discontinuing rofecoxib and instituting parenteral rehydration, the bleeding and diarrhea stopped. Endoscopic follow-up revealed regenerating mucosa in the transverse colon. The time relation, the absence of other causes of hemorrhage, and the clinical evolution all strongly support the probability of a causal relation between rofecoxib and hemorrhagic colitis. This case may raise awareness concerning the possibility of colonic lesions related to the new COX-2 inhibitors, similar to what is known about nonselective antiinflammatory agents. PMID- 11907362 TI - Crohn's disease and Takayasu's arteritis. AB - Takayasu's arteritis and inflammatory bowel disease are rarely found together in the reported literature. In particular, only one African-American patient with both Crohn's disease and Takayasu's arteritis has been reported. We present a case of an African-American patient previously diagnosed with Crohn's disease who subsequently developed Takayasu's arteritis and then presented with acute rectal bleeding. The differential diagnosis, diagnostic evaluation, and possible common pathophysiologic mechanism between the two diseases are discussed. PMID- 11907363 TI - Fatal hyperphosphatemia from a phosphosoda bowel preparation. AB - Oral phosphosoda is increasingly being used as a bowel preparation for colonoscopy, as it requires that a much smaller volume be ingested and is equally effective and less costly than polyethylene glycol-based electrolyte solutions. Oral phosphosoda has a good safety record, but complications of its use may occur. We describe a patient who died as a result of severe hyperphosphatemia after an oral phosphosoda bowel preparation. A 55-year-old man was admitted with rectal bleeding, abdominal pain, and vomiting. He had a history of diabetes, hypertension, and end-stage renal disease and had successful renal transplant 3 years prior. His initial serum creatinine, calcium, phosphate, and electrolyte levels were normal. He vomited after polyethylene glycol-based electrolyte solution, and an alternate bowel preparation with oral phosphosoda was recommended. He received 90 mL of oral phosphosoda as a single dose. Six hours later, he had cardiorespiratory arrest and was found to have hyperphosphatemia (serum phosphate, 17.8 mg/dL), a high anion gap acidosis, hypoxia, and oliguric renal failure. Resuscitation was unsuccessful. Autopsy showed ischemic colitis. We conclude that bowel preparation with phosphosoda may be associated with severe complications and should be avoided if there is any suggestion of impaired renal function or poor gut motility. PMID- 11907364 TI - Laboratory diagnostic tests in acute pancreatitis. AB - The diagnosis of acute pancreatitis depends on a combination of clinical assessment and laboratory testing. Although the serum amylase is the cornerstone laboratory test used in establishing the diagnosis of acute pancreatitis, there are limitations in the sensitivity and specificity that may be important for the clinician to recognize. The serum lipase level may be especially useful in patients with alcohol-induced acute pancreatitis. A new urinary test strip that uses trypsinogen-2 may have a role in establishing the diagnosis of acute pancreatitis. In addition, several new laboratory tests and new interpretations of old laboratory tests may assist in establishing the etiology and severity of acute pancreatitis. This review summarizes important aspects of standard laboratory tests and novel laboratory approaches in establishing the diagnosis, etiology, and severity of acute pancreatitis. PMID- 11907365 TI - Hepatic neoplasms: computed tomography and magnetic resonance features. AB - Over the last decade, major advances in computed tomography and magnetic resonance technology have occurred. These advances enable accurate, noninvasive detection and characterization of many hepatic neoplasms. This article illustrates the role of imaging in the evaluation of hepatic neoplasms and reviews the typical imaging features of both benign and malignant hepatic tumors. Benign tumors discussed include hemangiomas, focal nodular hyperplasia, hepatocellular adenoma, and simple cysts, as well as cysts associated with polycystic liver disease. Malignant neoplasms reviewed include metastases and conventional hepatocellular carcinoma as well as less common tumors such as fibrolamellar hepatocellular carcinoma, intrahepatic cholangiocarcinoma, angiosarcoma, and epithelioid hemangioendothelioma. PMID- 11907366 TI - Long-term follow-up of patients with acute hypertriglyceridemia-induced pancreatitis. AB - BACKGROUND: An acute and potentially life-threatening complication of hypertriglyceridemia (HTG) is acute pancreatitis (AP). Hypertriglyceridemia, usually severe, may be primary in origin or secondary to alcohol abuse, diabetes mellitus, pregnancy, and use of drugs. STUDY: The efficacy of treatment to prevent relapses in 17 patients with AP attributed to HTG was investigated in the current prospective study. The mean follow-up period of patients was 42 months. Hypertriglyceridemia-induced AP comprised 6.9% of all patients with AP (n = 246) hospitalized in our clinic during the study (6 years). RESULTS: Causative conditions of HTG-induced AP were familial HTG in eight patients, HTG caused by uncontrolled diabetes mellitus in five, HTG aggravated by drugs in two (one by tamoxifen and one by fluvastatin), familial hyperchylomicronemia (HCM) in one, and lipemia of pregnancy in one. During the acute phase of pancreatitis, patients underwent standard treatment. Thereafter, HTG was efficiently controlled with high dosages of fibrates or a fibrate plus acipimox, except for the patient with HCM, who was on a specific diet (the only source of fat was a special oil consisting of medium chain triglyceride) and taking a high dosage of acipimox. One of the patients died during the acute phase of pancreatitis with acute respiratory distress syndrome. During follow-up, maintenance treatment was successful and only one patient relapsed, because he discontinued diet and drug treatment. CONCLUSION: Appropriate diet and drug treatment, including dose titration, of severe HTG is very effective in preventing relapses of HTG-induced AP. PMID- 11907367 TI - Viral hepatitis and other infectious diseases in a homeless population. AB - GOALS: To determine the prevalence of four common infectious diseases-hepatitis B, hepatitis C, human immunodeficiency virus (HIV), and tuberculosis-as well as co-infection rates and risk factors in a homeless population. BACKGROUND: The prevalence of infectious diseases, especially viral hepatitis, among the homeless population is largely unknown. STUDY: This study consists of a retrospective analysis of the history and laboratory data collected from all homeless veterans admitted to a Veterans Administration (VA) domiciliary from May 1995 to March 2000. RESULTS: Of the homeless veterans admitted to a VA domiciliary program, 597 of 829 were screened for markers of all four infectious diseases. The overall prevalence of anti-hepatitis C virus (HCV) antibody, and positive result for purified protein derivative (PPD), anti-HIV antibody, and hepatitis B surface antigen (HbsAg) were 41.7%, 20.6%, 1.84% and 1.17%, respectively. At least one of the four markers was positive in 52.6% and more than one in 12%. Co-infection with HCV occurred commonly in veterans who were positive for anti-HIV (72.7%) and HBsAg (57.1%). Four self-reported major risk factors (intravenous drug use, alcohol abuse, previous imprisonment, and prior stay in a shelter) were evaluated. Multivariate analysis indicates that intravenous drug use and anti-HBs reactivity are independent risk factors for HCV infection, HCV infection for anti hepatitis B surface antibody reactivity, and older age for PPD positivity. CONCLUSIONS: Chronic hepatitis C and co-infections are common among the homeless population. Patients infected with HIV and hepatitis B virus frequently are co infected with HCV. Infections frequently are associated with certain identifiable risk factors. PMID- 11907368 TI - Arterioportal fistula causing acute pancreatitis and hemobilia after liver biopsy. AB - We report the case of a 49-year-old patient who developed hemobilia and acute pancreatitis from an arterioportal fistula after a percutaneous liver biopsy, and we analyze diagnostic testing and management based on a concise review of the available literature. Hemobilia can present as late as 10 days after liver biopsy. Acute pancreatitis is a rare complication of hemobilia. To our knowledge, this is the first documented case of an arterioportal fistula after percutaneous liver biopsy with the late manifestation of hemobilia and acute pancreatitis. PMID- 11907369 TI - Acute myeloid leukemia presenting as obstructive jaundice. AB - A 36-year-old man presented with abdominal pain and jaundice. Ultrasound examination of the abdomen showed dilatation of the intrahepatic bile ducts and a normal common bile duct. No calculi were demonstrated. A computerized tomography scan did not show any masses. A peripheral blood smear was diagnostic of acute myeloid leukemia. After remission-inducing chemotherapy, there was a complete regression of the jaundice, which had not recurred at the 12-month follow-up. PMID- 11907370 TI - Eosinophilic enterocolitis in a patient with rheumatoid arthritis. PMID- 11907371 TI - Infliximab treatment of an esophagobronchial fistula in a patient with extensive Crohn's disease of the esophagus. PMID- 11907372 TI - Transjugular intrahepatic portosystemic shunt for portal decompression before abdominal and retroperitoneal surgery in patients with severe portal hypertension. PMID- 11907373 TI - Gastric antral vascular ectasia as the only presenting feature of limited cutaneous systemic sclerosis. PMID- 11907374 TI - Multiple pseudoaneurysms associated with acute pancreatitis. PMID- 11907375 TI - Recurrence of Henoch-Schonlein purpura in association with colitis. PMID- 11907376 TI - Acute hepatitis associated with cetirizine intake. PMID- 11907380 TI - Epstein-barr virus can infect B-chronic lymphocytic leukemia cells but it does not orchestrate the cell cycle regulatory proteins. AB - OBJECTIVES: To understand the mechanism for the refractoriness of B-chronic lymphocytic leukemia (B-CLL) cells for Epstein-Barr virus (EBV)-induced immortalization. STUDY DESIGN/METHODS: Cultures were initiated with EBV-infected tonsillar B and B-CLL cells. Expression of EBNA-2 and some of the key players regulating G1/S phase transition such as c-myc expression, phosphorylation of Rb protein, expression of G1 cyclins, and the cyclin-dependent kinase inhibitor p27 were followed. RESULTS: In line with earlier studies, EBV infection induced c-myc expression, pRb phosphorylation, D2 and D3 expression, and disappearance of p27 in normal B cells. In contrast, EBV-infected B-CLL cells remained resting and they did not express c-myc; cyclin D2, ppRb and cyclin D3 were seen only in occasional cells. Importantly, p27 expression was maintained. CONCLUSIONS: In B CLL cells, the expression of the EBV-encoded nuclear proteins EBNAs is not followed by entrance to the cell cycle. Thus, the difference in the interaction of EBV-normal B cells and EBV-B-CLL cells is already apparent early after infection. PMID- 11907381 TI - Detection and quantification of multiple drug resistance mutations in HIV-1 reverse transcriptase by an oligonucleotide ligation assay. AB - OBJECTIVES: To develop an assay for the early detection and quantification of minor human immunodeficiency virus-1 populations bearing multiple drug resistance (MDR) mutations. STUDY DESIGN/METHODS: The oligonucleotide ligation assay (OLA) is based on ligation of probe and detector oligonucleotides annealed to a polymerase chain reaction amplicon strand with detection by an enzyme immunoassay. In OLA-MDR, oligonucleotides were designed to detect MDR mutations. The method was validated with wild-type and MDR mutant clones mixed at different proportions. RESULTS: K103N mutants were detected as minor populations (5%-30%) by OLA in 6 of 18 samples from patients treated with nonnucleoside reverse transcription inhibitors and classified as wild type by sequencing. In one patient, the kinetics of the increase of MDR mutants could be followed in sequential samples, with K103N being detected earlier by OLA than by sequencing. Q151M mutants were detected as minor populations (13%-24%) by OLA but not by sequencing in 4 samples. CONCLUSIONS: Oligonucleotide ligation assay MDR exhibits higher sensitivity than sequencing for detection of minor MDR mutant populations. PMID- 11907382 TI - Influence of human immunodeficiency virus type 1 envelope glycoprotein YXXL endocytosis/polarization signal on viral accessory protein functions. AB - INTRODUCTION: A tyrosine-based targeting signal in the intracytoplasmic domain of human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is required for basolateral targeting of viral budding in polarized epithelial cells, concentration of viral assembly at one pole of infected lymphocytes, and rapid endocytosis of the glycoprotein from the plasma membrane. In HIV-1, the process of viral assembly and budding is complex and involves the participation of viral accessory proteins. STUDY DESIGN/METHODS: In this study, we examined whether the functions of the Nef, Vif, and Vpu proteins can influence or be influenced by the presence of envelope targeting signal in epithelial cells or lymphocytes. A series of proviral DNAs combining a mutation that inactivates the targeting signal with independent mutations in the three accessory proteins was constructed for this purpose. RESULTS: It was found that none of these three accessory proteins affected the basolateral release of the virus in polarized MDCK cells. Reciprocally, a mutation abolishing targeting of the viral envelope glycoprotein did not affect the phenotype conferred by the accessory proteins. Interestingly, the mutation abolishing targeting increased viral infectivity only in the presence of the Vpu protein. This phenotype was found to be associated with the release-enhancing function of Vpu and with an increased incorporation of viral envelope glycoprotein in virions. CONCLUSIONS: These data clearly show that accessory protein functions, and more specifically Vpu modulation of viral infectivity, can be affected by variations in the viral envelope glycoprotein. PMID- 11907383 TI - Diversity of HIV-1 subtype E in semen and cervicovaginal secretion. AB - OBJECTIVES: To characterize human immunodeficiency virus type 1 (HIV-1) subtype E variants in blood and genital fluid of infected Thai couples. STUDY DESIGN/METHODS: Blood and genital fluid were collected from 30 asymptomatic healthy HIV-1 subtype E infected couples from Bangkok, Thailand from 1995 to 1998. RESULTS: All 60 viruses in blood samples were identified as subtype E by heteroduplex mobility assay. The biotype of viruses founded in blood was syncytium-inducing (SI), whereas M-tropic and non-syncytium-inducing (NSI) isolates were predominantly detected in genital fluid. HIV-1 proviral DNA was detected in 43.33% and 56.67%, and viral RNA was detected in 93.33% and 56.67%, of semen (n = 30) and cervicovaginal secretion (n = 30) samples tested, respectively. A higher intersample genetic distance and more positive charge of the V3 loop were found in blood strains composed of genital fluid strains (22.30 +/- 5.92% and 17.96 +/- 6.3%), which was statistically significant (P = 0.003). The env V1-V4 intraperson variation of the HIV-1 subtype E in the blood and genital fluid of each individual was in the range 3.0%-5.7%. We also determined the intrasample variation of HIV-1 from blood and genital fluid by heteroduplex mobility assay. The mean heteroduplex mobility of the HIV-1, V1-V4 region of env gene, in blood (n = 8) and genital fluid (n = 8) was 0.59 +/- 0.06 and 0.74 +/- 0.11 (t test, p = 0.001), respectively. CONCLUSIONS: There was genetic and phenotypic compartmentalization of HIV-1 subtype E in blood and genital fluid with the presence of SI and NSI phenotypic variants as a common property of subtype E isolates from blood and genital fluid, respectively. PMID- 11907384 TI - HIV-1 IgA specific serum antibodies and disease progression during HIV-1 infection. AB - OBJECTIVE: To evaluate the role of serum human immunodeficiency virus type 1 immunoglobulin A (HIV-1 IgA) antibodies in the progression of HIV-1 infection in relation to viral load and CD4 cell counts. METHODS: Sequential serum specimens were obtained from 218 homosexual men: 123 HIV-1 seropositives, 24 HIV-1 seroconverters, and 71 HIV-1 seronegatives. HIV-1 IgA antibodies were tested blindly by enzyme-linked immunosorbent assay and Western blot. T-lymphocyte subsets were measured by flow cytometry. Viral plasma load was determined by a sensitive branched DNA assay. RESULTS: HIV-1 IgA antibodies with a titer greater than or equal to 50 were detected among 50% of the seroconverters, 27% of the HIV 1-seropositive asymptomatic subjects, 25% of lymphadenopathy, and 23% of HIV-1 related symptomatic subjects. Among patients with the acquired immune deficiency syndrome, the prevalence of virus-specific IgA antibodies (55%) was significantly higher (p < 0.03) as compared with the HIV-1-seropositive asymptomatic subjects, lymphadenopathy and HIV-1-related symptomatic patients, but not versus the seroconverters (p = 0.8). IgA antibodies to HIV-1 gP160 were the most prevalent among all subjects tested. A significant decrease in CD4 cell counts was observed after HIV-1 seroconversion. Viral load was slightly higher among the seroconverters who demonstrated higher (> or =50) HIV-1 IgA levels. CONCLUSIONS: HIV-1 IgA serum antibodies did not predict the progression of the disease. Correlation between HIV-1 IgA antibodies titer, viral load, and CD4 cell counts was not detected. PMID- 11907385 TI - Assessment of a rapid HIV test strategy during labor: a pilot study from Rio de Janeiro, Brazil. AB - OBJECTIVES: To use two rapid human immunodeficiency virus (HIV) tests at labor, measure test acceptance and performance, and measure HIV prevalence in these women. METHODS: Between February and October 2000, two rapid tests (Determine; Abbott, Chicago, IL, U.S.A. and Double Check; Orgenics, Yavne, Israel) were used in three public maternities in Rio de Janeiro, Brazil. Enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) analysis confirmed positive and discordant results. RESULTS: Of the 858 patients who were enrolled, the mean gestational age was 36 weeks (median = 39, mode = 40) and 17 (2%) refused testing. Of the 841 patients tested, 13 were positive by both tests, which represents a 1.5% prevalence (95% confidence interval: 0.7%-2.3%); all were confirmed by ELISA and WB analysis. Seven samples gave discordant results by the rapid tests; of these, six were ELISA-negative/WB-negative and one was ELISA negative/WB-indeterminate. The positive predictive value for samples that were positive by both rapid tests simultaneously was 100%. CONCLUSIONS: Two rapid HIV tests used at labor were well accepted (98%). When the combined results of the two rapid tests (but not a single rapid test) were analyzed, this strategy was as efficient as the standard ELISA and WB HIV strategy for correctly classifying individuals. PMID- 11907386 TI - Human papillomavirus absence predicts normal cervical histopathologic findings with abnormal papanicolaou smears: a study of a university-based inner city population. AB - INTRODUCTION: We studied the role of human papillomavirus (HPV) typing in predicting cervical dysplasia in women with abnormal Papanicolaou (Pap) test results. STUDY DESIGN/METHODS: A university colposcopy clinic-based consecutive sample of 179 women completed a questionnaire and underwent colposcopy, HPV typing (Hybrid Capture System HPV DNA Assay II; Digene Diagnostics, Gaithersburg, MD, USA), and biopsy (if indicated). RESULTS: No severe dysplasia was observed in women with low-risk HPV or in women with negative HPV test results who had a low grade abnormality on the Pap test. High-risk (HR) HPV was present in every case of severe dysplasia on biopsy. The cumulative odds risk for cervical dysplasia was 1.11 in HIV(+) women with low-grade squamous intraepithelial lesion on the Pap test who were older than 21 years of age and HPV-HR(+). CONCLUSIONS: In the population studied, HPV typing is a valuable adjunct to a low-grade abnormality on the Pap test in predicting the absence of cervical dysplasia on biopsy. Larger prospective population-based studies are needed to study the role of HPV as a negative predictor of disease in cervical dysplasia. PMID- 11907387 TI - Combined administration of diltiazem and nicardipine attenuates hypertensive responses to emergence and extubation. AB - Diltiazem and nicardipine, when injected as a mixture during anesthesia, reduce blood pressure in an additive manner without changing heart rate. The author evaluated the use of this mixture for controlling the blood pressure during emergence from general anesthesia and at extubation. The subjects included 15 preoperative hypertensive (HT) patients who underwent various types of surgery and 18 patients with subarachnoid hemorrhage (SAH) who underwent clipping of a cerebral aneurysm. General anesthesia was maintained with isoflurane or sevoflurane, supplemented with fentanyl. A mixed solution containing 2.5 mg diltiazem plus 0.5 mg nicardipine in 1 mL was injected intermittently every 2 to 4 minutes to bring the blood pressure to its resting level from cessation of inhaled anesthetics to extubation. Untreated patients who underwent similar types of surgery and anesthesia were selected for comparison. The average systolic blood pressure during emergence and at extubation increased to 156 +/- 19 mm Hg (mean +/- standard deviation) and 170 +/- 10 mm Hg in the untreated HT group, and increased to 157 +/- 16 mm Hg and 170 +/- 5mm Hg in the untreated SAH group. Systolic blood pressure was well controlled at 127 +/- 14 mm Hg and 145 +/- 14 mm Hg in the treated HT group with 3.7 +/- 1.9 mL of the mixture, and at 120 +/- 9 mm Hg and 137 +/- 20 mm Hg in the treated SAH group with 7.1 +/- 2.5 mL of the mixture. No significant difference (P < .05) in the heart rate was found between the untreated and the treated HT or SAH groups. Two patients in the treated SAH group exhibited tachycardia. The combined administration of diltiazem and nicardipine can help control blood pressure in patients with a possible HT response to emergence from general anesthesia and extubation. PMID- 11907389 TI - A randomized, double-blind comparison of ondansetron versus placebo for prevention of nausea and vomiting after infratentorial craniotomy. AB - Ondansetron was compared with placebo for nausea and vomiting prophylaxis after fentanyl/isoflurane/relaxant anesthesia and infratentorial craniotomy. Eight milligrams intravenous ondansetron or vehicle was administered at skin closure. Nausea, emesis, and antiemetic use were recorded at 0, 0.5, 1, 4, 8, 12, 24, and 48 hours. There were no significant intergroup differences for nausea incidence at any interval, but cumulatively the placebo group was 3.2 times more likely to develop nausea during the first 12 hours (P = .04). Nausea incidence was bimodal in both groups, peaking during the first 1 to 4 hours. A nadir occurred at 8 to 12 hours, but nausea increased during the next 36 hours. By 48 hours, approximately 40% of patients in both groups were still nauseated. Reduced vomiting frequency was seen with ondansetron at 4, 8, 12, and 24 hours (P < .05). Despite rescue antiemetics, emesis occurred in an irregular pattern with episodes still observed in 35% of placebo patients at 48 hours. For ondansetron, emesis was infrequent for the first 12 hours but then a persistent increase was observed (48 hours, 22%). The incidence of rescue antiemetic use was 65% for both groups. There was no effect of gender. Nausea and vomiting are frequent and protracted after infratentorial craniotomy. Administration of single-dose ondansetron (8 mg intravenously) at wound closure was partially effective in reducing acute nausea and vomiting but had little delayed benefit. Scheduled prophylactic administration of antiemetic therapy during the first 48 hours after infratentorial craniotomy should be evaluated for efficacy and safety. PMID- 11907388 TI - Postoperative pain management after supratentorial craniotomy. AB - The aim of this study was to compare the analgesic efficacy of three different postoperative treatments after supratentorial craniotomy. Sixty-four patients were allocated prospectively and randomly into three groups: paracetamol (the P group, n = 8), paracetamol and tramadol (the PT group, n = 29), and paracetamol and nalbuphine (the PN group, n = 27). General anesthesia was standardized with propofol and remifentanil using atracurium as the muscle relaxant. One hour before the end of surgery, all patients received 30 mg/kg propacetamol intravenously then 30 mg/kg every 6 hours. Patients in the PT group received 1.5 mg/kg tramadol 1 hour before the end of surgery. For patients in the PN group, 0.15 mg/kg nalbuphine was injected after discontinuation of remifentanil, because of its mu-antagonist effect. Postoperative pain was assessed in the fully awake patient after extubation (hour 0) and at 1, 2, 4, 8, and 24 hours using a visual analog scale (VAS). Additional tramadol (1.5 mg/kg) or 0.15 mg/kg nalbuphine was administered when the VAS score was > or = 30 mm. Analgesia was compared using the Mantha and Kaplan-Meier methods. Adverse effects of the drugs were also measured. The three groups were similar with respect to the total dose of remifentanil received (0.27 +/- 0.1 mircog/kg/min). In all patients, extubation was obtained within 6 +/- 3 minutes after remifentanil administration. Postoperative analgesia was ineffective in the P group; therefore, inclusions in this group were stopped after the eighth patient. Postoperative analgesia was effective in the two remaining groups because VAS scores were similar, except at hour 1, when nalbuphine was more effective (P = .001). Nevertheless, acquiring such a result demanded significantly more tramadol than nalbuphine (P < .05). More cases of nausea and vomiting were observed in the PT group but the difference was not significant (P < .06). In conclusion, pain after supratentorial neurosurgery must be taken into account, and paracetamol alone is insufficient in bringing relief to the patient. Addition of either tramadol or nalbuphine to paracetamol seems necessary to achieve adequate analgesia, with, nevertheless, a larger dose of tramadol to fulfill this objective. PMID- 11907391 TI - Antioxidant actions and early ultrastructural findings of thiopental and propofol in experimental spinal cord injury. AB - Thiopental and propofol are effective antioxidant agents. The current study was undertaken to examine the neuroprotective effects of a single intraperitoneal dose of thiopental and propofol. Effects of the drugs were evaluated by lipid peroxidation and ultrastructural findings. Fifty male Wistar rats were divided into five groups. Group 1 was the control group. Rats underwent laminectomy only, and nontraumatized spinal cord samples were obtained 1 hour after surgical intervention. All other rats sustained a 50-g/cm contusion injury by the weight drop technique. Group 2 rats underwent spinal cord injury alone, group 3 rats received 1 mL intralipid solution intraperitoneally immediately after trauma as the vehicle group, group 4 rats received a 15-mg/kg single dose of thiopental, and group 5 rats received a 40-mg/kg single dose of propofol intraperitoneally following the trauma. Samples from groups 2, 3, 4, and 5 were obtained 1 hour after injury. Lipid peroxidation was determined by measuring the concentration of malondialdehyde in the spinal cord tissue. The ultrastructure of the spinal cord was determined by electron microscopy. The contusion injury was associated with a rise in lipid peroxidation. Compared with the trauma group there was significant attenuation in lipid peroxidation of groups 4 and 5. Ultrastructural findings showed that the rats of group 4 sustained minor damage after spinal cord injury, but there was more evident damage in group 5 rats. These results indicate that thiopental decreases lipid peroxidation and improves ultrastructure, whereas propofol decreases lipid peroxidation without improving ultrastructure 1 hour after spinal cord injury in rats. PMID- 11907390 TI - Intra-arterial 133Xe measurements suggest a dose-dependent increase in cerebral blood flow during intracarotid infusion of adenosine in nonhuman primates. AB - Intra-arterial vasodilators, such as papaverine, have been used to treat cerebrovascular insufficiency. The short biologic half-life, and the vasodilating and neuroprotective properties of adenosine could be useful during the treatment of cerebral ischemia. However, in human subjects a proposed intracarotid dose of 1 mg/min adenosine was ineffective in augmenting cerebral blood flow (CBF). The object of this experiment was to determine the dose-CBF response characteristics of intracarotid adenosine in nonhuman primates. Studies were conducted on five male baboons under isoflurane anesthesia. After transfemoral internal carotid artery cannulation, changes in CBF (intra-arterial 133Xe technique) were determined after intracarotid infusion of saline and three increasing doses of adenosine (0.5, 1.0, and 1.5 mg/min). Each infusion lasted 5 minutes. Data (mean +/- standard deviation) were analyzed by repeated-measure analysis of variance and the post hoc Tukey test. Intracarotid adenosine (0.5, 1.0, and 1.5 mg/min) resulted in a dose-dependent increase in CBF from 22.6 +/- 4 mL/100 g/min at baseline to 50 +/- 15, 65 +/- 22, and 83 +/- 31 mL/100 g/min respectively (n = 5, P < .05 each). No adverse hemodynamic side effects were noted, and animals recovered promptly from anesthesia. The authors conclude that intracarotid adenosine in the range of 0.5 to 1.5 mg/min results in a robust increase in CBF. Based on body weight, intracarotid adenosine in a dose range of 2.5 to 7.5 mg/min may be required to augment CBF in human subjects. PMID- 11907392 TI - Characterization of the cerebral blood flow response to balloon deflation after temporary internal carotid artery test occlusion. AB - The authors tested the hypothesis that cerebral blood flow (CBF) would increase after acute and relatively brief internal carotid artery (ICA) test occlusion, and examined the relationship of the postdeflation CBF to the development of neurologic symptoms. In 16 patients undergoing ICA test occlusion with deliberate hypotension, the authors measured intracarotid 133Xe CBF at baseline, occlusion, and deflation. Four patients developed neurologic symptoms during occlusion. As positive controls, 11 other patients received intracarotid verapamil or papaverine before deflation as part of another protocol. Balloon occlusion was 23.1 +/- 10.5 minutes (mean +/- standard deviation) in duration. At 1.3 +/- 1.6 minutes after balloon deflation, there was a trend (12 +/- 31%) for CBF to increase (48 +/- 9 mL/100 g/min versus 53 +/- 17 mL/100 g/min, P =.15), and a 16 +/- 27% decrease in cerebrovascular resistance (CVR; 2.1 +/- 0.6 mm Hg/100 g/min/mL versus 1.7 +/- 0.6 mm Hg/100 g/min/mL, P =.03) compared with baseline values. By comparison, patients who received a intracarotid dilator demonstrated a 53 +/- 55% increase in CBF (48 +/- 10 mL/100/min versus 70 +/- 23 mL/100 g/min, P = .007) and a 33 +/- 31% decrease in CVR (2.2 +/- 0.6 mm Hg/100 g/min/mL versus 1.4 +/- 0.6 mm Hg/100 g/min/mL, P = .0007) compared with baseline. Analysis of variance and regression analysis showed no other relationships between postocclusion CBF and balloon occlusion duration, distal internal carotid occlusion ("stump") pressure, or the development of neurologic symptoms. Acute, temporary interruption of ICA blood flow resulted in minimal postocclusive changes in cerebrovascular hemodynamics, even in those patients who developed neurologic symptoms during the period of test occlusion. PMID- 11907393 TI - Jugular venous oxygen saturation thresholds in trauma patients may not extrapolate to ischemic stroke patients: lessons from a preliminary study. AB - The authors' first examinations of 10 patients with severe hemispheric stroke indicate that bedside monitoring of cerebral blood flow (CBF) is of clinical value as a prognostic tool for outcome and as therapy of elevated intracranial pressure (ICP). Jugular venous oximetry, which is easier to handle and provides on-line data, may also be of prognostic value in patients with ischemic stroke. No clinical studies are available on patients with hemispheric infarctions. Therefore, in a second data analysis from the same patient population, the authors' objective was to estimate the clinical value of monitoring cerebral hemodynamics and metabolism with jugular bulb catheters in treatment of severe postischemic brain edema. In 10 patients with severe hemispheric infarctions, ICP, jugular venous oxygen saturation (SjvO2), CBF, and cerebral metabolic rate of oxygen (CMRO2) were measured prospectively. A total of 101 ICP, SjvO2, and 92 CBF measurements were obtained. Only two SjvO2 values were below the critical thresholds to detect secondary ischemic events defined in trauma patients (SjvO2 < 50%). Intracranial pressure elevations more than 20 mm Hg and pupillary disturbances were treated with osmotherapy (mannitol or hypertonic NaCl hydroxyethyl starch solution) or mild hyperventilation in combination with tromethamine-buffer. In 8 of 17 pairs of measurements with treated elevated ICP, CMRO2 varied and changes of SjvO2 did not reflect changes in CBF. Jugular bulb oximetry should interpreted with caution in patients with severe hemispheric infarction. Critical thresholds defined in trauma patients may not be extrapolated to ischemic stroke. PMID- 11907394 TI - Aprotinin and deep hypothermic cardiopulmonary bypass with or without circulatory arrest for craniotomy. AB - Deep hypothermic cardiopulmonary bypass with or without circulatory arrest has been used to facilitate the surgical repair of complex cerebrovascular lesions. The advantages of deep hypothermia have been tempered by the occurrence of coagulopathy that is associated with substantial morbidity and mortality. This study analyzed retrospectively the records of 13 patients who underwent cerebrovascular neurosurgery using deep hypothermic cardiopulmonary bypass with or without circulatory arrest during the period 1993 through 1999. All patients received the serine protease inhibitor aprotinin in an effort to avoid the development of a coagulopathy, defined as hemorrhage requiring reoperation. No patients developed postoperative intracranial hemorrhage. There was also no evidence of renal dysfunction, deep venous thrombosis, myocardial infarction, or pulmonary embolism. In conclusion, this study suggests that aprotinin may be beneficial to avoid the coagulopathy that is more likely to occur if deep hypothermic cardiopulmonary bypass with or without circulatory arrest is used for craniotomy without adverse effects on renal function or apparent thrombotic complications. PMID- 11907395 TI - Effects of hypothermia on median nerve somatosensory evoked potentials during spontaneous circulation. AB - Perioperative-induced hypothermia is a common means of reducing ischemic injury in neurosurgical procedures and cardiac surgery, and it may occur accidentally. Somatosensory evoked potentials (SSEPs) are used frequently for neurophysiologic monitoring of these procedures. The effects of hypothermia on SSEPs have been studied widely in humans with cardiopulmonary bypass (CPB) during nonpulsatile flow. However, changes of latency and amplitude of early SSEP components during spontaneous circulation have not yet been studied. Median nerve SSEPs were recorded in 21 patients during rewarming from 32 to 36 degrees C core temperature. Latencies and amplitudes of N9, N13, N20, and central conduction time were registered at 32, 34, and 36 degrees C. Latencies of N9, N13, and N20 were prolonged at 32 degrees C compared with 36 degrees C (N9: 13.4 +/- 1.4 msec versus 11.8 +/- 1.4 msec, P <.05; N13: 17.6 +/- 1.9 msec versus 15.4 +/- 1.4 msec, P <.01; N20: 26.5 +/- 1.8 msec versus 22.4 +/- 1.6 msec, P <.001). Amplitude of N20 was higher at 32 degrees C compared with 36 degrees C (2.86 +/- 1.94 microV versus 2.07 +/- 1.47 microV, P < .05). Central conduction time decreased by 27%, and peripheral latency of N13 decreased by 14%. The increase in SSEP latency (N9, N13, and N20) and central conduction time during moderate hypothermia of 32 degrees C and spontaneous circulation are comparable with those during nonpulsatile flow on CPB. In contrast to nonpulsatile flow, the amplitude of N20 was increased significantly (P < .05) during moderate hypothermia and pulsatile circulation. These results suggest to be cautious about generalizing the effects of hypothermia on SSEP during CPB to spontaneous circulation. PMID- 11907396 TI - Dynamic left ventricular outflow obstruction during lumbar laminectomy as an unexpected cause of intraoperative hypotension. AB - We present a case of previously undiagnosed hypertrophic obstructive cardiomyopathy (HOCM) with left ventricular outflow obstruction in a woman anesthetized for lumbar hemilaminectomy and diskectomy. The treatment of her sudden unexplained hypotension was initially confounded by a diagnosis of compensated congestive heart failure and diuretic therapy. Swift intervention with transesophageal echocardiography revealed the tru pathology altering her intraoperative treatment and her subsequent chronic treatment for her heart condition. PMID- 11907397 TI - Usefulness of monitoring brain tissue oxygen pressure during awake craniotomy for tumor resection: a case report. AB - Awake craniotomy is indicated for surgical resection of tumors located near eloquent areas of the brain. The anesthetic technique is based on a combination of local anesthesia, sedation, and analgesia. Usually only clinical parameters are assessed and no other cerebral oxygenation monitoring techniques are applied. The authors report the use of brain tissue oxygen pressure monitoring during awake craniotomy. A 48-year-old right-handed man with a left temporoparietal mass was scheduled for awake craniotomy, cortical stimulation, and selective tumor removal. Monitoring included electrocardiography, pulse oximetry, end-tidal CO2, bladder temperature, invasive and noninvasive arterial pressure, and brain tissue oxygen pressure (PtiO2). The anesthetic technique consisted of continuous perfusions of 0.02 to 0.05 microg/kg/min remifentanil, propofol (target concentration, 0.5 to 1.2 microg/mL), and 25 to 50 microg/kg/min esmolol, and local anesthetic blockade of the head pin insertion sites and surgical incision area (a mixture of 0.2% ropivacaine, 1% lidocaine, and epinephrine, 1:200 000). Intraoperative cortical stimulation was performed to guide the resection according to the patient's verbal response. A change in PtiO2 was observed, gradually falling from 28 mm Hg at the beginning of the intervention down to 3 mm Hg. At this stage, surgical resection was concluded. On arrival at the intensive care unit, mixed dysphasia and slight weakness of the right arm were noted. Three weeks after surgery, the patient's speech is improving and the motor deficit has disappeared. This case suggests a possible role of PtiO2 in awake craniotomy as an aid in detecting intraoperative adverse events, but further experience with PtiO2 in this setting is needed. PMID- 11907398 TI - Should one rely on capnometry when a capnogram is not seen? AB - Capnography is one of the basic monitoring techniques in day-to-day anesthesia practice that provides information not only regarding the patient's ventilation, circulation, and metabolism, but also regarding proper functioning of a closed circle system. The authors report a case in which after endotracheal intubation the end-tidal capnometric reading rose very high, but the capnogram was not seen on the monitor. The unexpectedly high capnometric reading with absent waveform during intermittent positive pressure ventilation without any apparent cause and consequent delayed institution of corrective measures resulted in severe brain bulge. There was severe hypercarbia as a result of a malfunctioning expiratory unidirectional valve that allowed rebreathing. Retrospective retrieval of data showed that a fraction of inspired carbon dioxide was also high and the baseline was raised beyond the usual range of 0 to 40 mm Hg, giving the impression of an absent waveform on the existing scale. In conclusion, one should keep in mind the possibility of expiratory valve malfunction resulting from dislodgment while wheeling the anesthesia machine, the view becoming obscured as a result of condensation of water vapor on the under surface of the plastic case, and one should rely on the capnometric reading unless proved otherwise. Thus, one can prevent potential hazards of rebreathing. PMID- 11907399 TI - Metoclopramide-induced raised intracranial pressure after head injury. AB - We report a case of raised intracranial pressure in a head-injured patient following the intravenous administration of metoclopramide. The patient required admission to an intensive care unit after a road traffic accident. A CT scan of the head was consistent with diffuse axonal injury and supportive management included intracranial pressure monitoring. On the third day after admission, intravenous metoclopramide 10mg was administered to aid gastric emptying during nasogastric feeding. Intracranial pressure increased to 39mmHg from a baseline of 15-20mmHg. The same dose of metoclopramide was repeated the next day during transcranial doppler studies with an increase in ICP to 34mmHg and an associated rise in middle cerebral artery systolic blood velocity from 122cms-1 to 150cms-1. This effect of metoclopramide has not been previously reported. PMID- 11907407 TI - Different effects of tacrolimus and cyclosporine on renal hemodynamics and blood pressure in healthy subjects. PMID- 11907408 TI - Presence of autoantibody against ATTR Val30 Met after sequential liver transplantation. PMID- 11907409 TI - C1 inhibitor for prophylaxis of xenograft rejection after pig to cynomolgus monkey kidney transplantation. PMID- 11907410 TI - Autoimmune and pregnancy complications in the daughter of a kidney transplant patient. PMID- 11907411 TI - Elimination of donor-specific alloreactivity prevents cytomegalovirus-accelerated chronic rejection in rat small bowel and heart transplants. AB - BACKGROUND: The primary cause for late failure of vascularized allografts is chronic rejection (CR) characterized by transplant vascular sclerosis (TVS). Cytomegalovirus (CMV) infection accelerates TVS and CR by unclear mechanisms involving direct effects of CMV, indirect effects of the recipient's immune response to CMV, or interactions between CMV and the recipient's alloreactivity. This study examined the role of CMV and the alloreactive response in the development of TVS using bone marrow chimerism (BMC) in rat small bowel (SB) and heart transplantation models. METHODS: Fisher 344 (F344) rat heart or SB grafts were transplanted into F344/Lewis bone marrow chimera. F344 heart or SB grafts transplanted into Lewis recipients (low-dose cyclosporine) were positive controls for the development of TVS. Lewis heart or SB grafts transplanted into Lewis recipients (+/-cyclosporine) were transplantation controls. The effect of rat CMV (RCMV) (5x105 plaque-forming units) on TVS (neointimal index, NI) and graft survival was studied in these groups. RCMV infection was assessed by serologic analysis and quantitative polymerase chain reaction techniques (TaqMan). RESULTS: RCMV infection accelerated the time to graft CR (SB 70-38 days; hearts 90-45 days) and increased the severity of TVS in both the SB allografts (day 38, NI=27 vs. 52) and the heart allografts (day 45, NI=43 vs. 83). Grafts from CMV-infected syngeneic recipients failed to develop TVS and CR. Donor-specific tolerance induced by BMC prevented allograft TVS and CR in both transplant models. In contrast to naive Lewis recipients, RMCV infection failed to cause allograft TVS and CR in bone marrow (BM) chimeras. CONCLUSIONS: The events in CMV-induced acceleration of TVS involve a crucial interplay between CMV infection and the recipient's alloreactive immune response. PMID- 11907412 TI - C1-inhibitor for prophylaxis of xenograft rejection after pig to cynomolgus monkey kidney transplantation. AB - BACKGROUND: Early rejection of discordant porcine xenografts in primate recipients is initiated by the intragraft binding of either preformed (hyperacute xenograft rejection) or induced (acute vascular rejection) antiporcine recipient antibodies with subsequent complement activation via the classical pathway. We have investigated the efficacy of the supplemental administration of C1-inhibitor (C1-INH), a specific inhibitor of the classical complement activation pathway, for prophylaxis of xenograft rejection in a pig to primate kidney xenotransplantation setting. METHODS: Based on the results of pharmacokinetic studies performed in two nontransplanted monkeys, supplemental C1-INH therapy was administered daily to three Cynomolgus monkeys receiving a life-supporting porcine kidney transplant together with cyclophosphamide-induction/cyclosporine A/mycophenolat-mofetil/steroid immunosuppressive therapy. RESULTS: In the three monkeys receiving porcine kidney xenografts and continuous C1-INH treatment none of the grafts underwent hyperacute rejection; all xenografts showed initial function. Recipient survival was 13, 15, and 5 days. No graft was lost due to acute vascular rejection. All animals died with a functioning graft (latest creatinine 96, 112, and 96 micromol/liter) due to bacterial septicemia. CONCLUSION: We conclude that, in our model, supplemental C1-INH therapy together with a standard immunosuppressive regimen can be helpful for prevention of xenograft rejection in a pig to primate kidney xenotransplantation setting. The optimal dose and duration of C1-INH treatment, however, has yet to be determined. PMID- 11907413 TI - A comparison of rat tracheal transplant models: implantation verses anastomotic techniques for the study of airway rejection. AB - BACKGROUND: In rodent models, investigators have transplanted donor tracheas into a recipient rat's abdomen or s.c. tissue to study airway rejection. We describe a modification of this model, which provides improved histology to study the airway injury related to obliterative bronchiolitis. METHODS: The standard technique of implanting the donor trachea was compared to a model in which a tracheal Y graft was created by anastomosis of the donor trachea to the recipient airway. Syngeneic and allogeneic tracheal grafts (Lewis and Brown Norway rats) were harvested at 2 and 4 weeks using each model (eight groups). RESULTS: Gross patency at the tracheal anastomosis grafts was 100%. All donor tracheas, which were implanted without an anastomosis, were occluded with mucus (syngeneic) or granulation tissue (allogeneic). Syngeneic implant grafts demonstrated significantly less lumenal granulation tissue 35.3%+/-32 than the allograft implant group (95.3%+/-9.2, P=0.0005 at 4 weeks). The anastomotic allograft group demonstrated significantly less lumenal granulation tissue 48.3%+/-23.7 when compared with the implanted allograft group (P=0.003). The implanted allograft demonstrated a severe loss of epithelial integrity by 2 weeks (16.7%+/-38), which progressed to complete loss by 4 weeks (P=0.0001 and P=0.0001 vs. native). This loss was significantly more than that of the anastomotic group at 2 weeks (89.5%+/-13, P=0.004) and 4 weeks (88.3+/-29, P=0.005). CONCLUSIONS: The rat tracheal allograft anastomosed to the recipient airway demonstrated less lumenal granulation tissue obstruction and better preservation of epithelial integrity than an implant allograft, suggesting that an open airway improves assessment of transplant-related changes associated with rejection. PMID- 11907414 TI - Advantages of normothermic perfusion over cold storage in liver preservation. AB - BACKGROUND: To minimize the ischemia-reperfusion injury that occurs to the liver with the current method of preservation and transplantation, we have used an extracorporeal circuit to preserve the liver with normothermic, oxygenated, sanguineous perfusion. In this study, we directly compared preservation by the standard method of simple cold storage in University of Wisconsin (UW) solution with preservation by perfusion. METHODS: Porcine livers were harvested from large white sows weighing between 30 and 50 kg by the standard procedure for human retrieval. The livers were preserved for 24 hr by either cold storage in UW solution (n=5) or by perfusion with oxygenated autologous blood at body temperature (n=5). The extracorporeal circuit used included a centrifugal pump, heat exchanger, and oxygenator. Both groups were then tested on the circuit for a 24 hr reperfusion phase, analyzing synthetic function, metabolic capacity, hemodynamics, markers of hepatocyte and reperfusion injury, and histology. RESULTS: Livers preserved with normothermic perfusion were significantly superior (P=0.05) to cold-stored livers in terms of bile production, factor V production, glucose metabolism, and galactose clearance. Cold-stored livers showed significantly higher levels of hepatocellular enzymes in the perfusate and were found to have significantly more damage by a blinded histological scoring system. CONCLUSIONS: Normothermic sanguineous oxygenated perfusion is a superior method of preservation compared with simple cold storage in UW solution. In addition, perfusion allows the possibility to assess viability of the graft before transplantation. PMID- 11907415 TI - Liposomal encapsulation significantly enchances the immunosuppressive effect of tacrolimus in a discordant islet xenotransplant model. AB - BACKGROUND: Encapsulation of tacrolimus (TAC) in a lipid bilayer to form liposome encapsulated tacrolimus (LTAC) alters the biodistribution profile, half-life, and efficacy in organ allotransplantation models. LTAC has not been applied to either cell transplantation or xenotransplantation. METHODS: To test the efficacy of LTAC in a discordant islet xenograft model, tilapia (fish) islets were transplanted under the left kidney capsules of streptozotocin-diabetic Balb/c mice. Recipient mice (groups I-VI) were treated with: I, untreated; II, empty liposomes; III, TAC (2 mg/kg/day); IV, TAC (5 mg/kg/day); V, LTAC (2 mg/kg/day); or VI, LTAC (5 mg/kg/day); all treatments were for 35 days or until rejection (i.e., two glucose measurements >200 mg/dl). Graft-bearing kidneys were removed for histology after rejection. RESULTS: Mean graft survival time (mGST) for control groups I and II were 6.7+/-1.4 (n=6) and 7.5+/-1.3 days (n=4), respectively. Daily TAC treatment at 2 mg/kg/d (III) did not prolong graft function (mGST=7.7+/-1.6; n=6) although 5 mg/kg/day (IV) produced minimal prolongation to 12.8+/-4.8 days (n=12). Treatment with LTAC at 2 mg/kg/day (V) significantly prolonged mGST to 26.6+/-4.9 (n=5); however, all recipients rejected during treatment (i.e.,<35 days). LTAC at 5 mg/kg/day (VI) further prolonged mGST to 39.9+/-11.8 days (n=12) with only one mouse rejecting before day 35. Histologically, at the time of functional rejection, grafts were generally either totally or partially effaced by mononuclear cells, eosinophils, and fibrosis. In groups VI, islet grafts removed from two mice that died while they were normoglycemic and from a mouse terminated while it was normoglycemic at day 36 were viable, well-granulated, and free from cellular infiltration. The group VI grafts examined at rejection (i.e., 1-2 weeks after discontinuing LTAC) were generally totally obliterated and were in two instances associated with nodular aggregates of atypical lymphocytes resembling posttransplant lymphoproliferative disorder. CONCLUSIONS: LTAC is the most potent immunosuppressive compound we have tested in our discordant fish-to-mouse islet xenograft model; however, toxicity is an issue at high doses. PMID- 11907416 TI - Fetal pig beta cells are resistant to the toxic effects of human cytokines. AB - BACKGROUND: The cytokine tumour necrosis factor-alpha (TNF-alpha) is thought to be responsible for primary nonfunction of islets when transplanted. This, and two other cytokines, interleukin-1beta (IL-1beta) and interferon-gamma (IFN-gamma) are also implicated in the autoimmune destruction of beta cells. It is unknown if the fetal pig beta cell, which is being transplanted as a treatment for type 1 diabetes, is affected by these cytokines. METHODS: We compared the effects of the cytokines on the function and viability of adult and fetal pig beta cells. The cells were cultured for up to 3 days in the presence of 2000 pg/ml of human IL 1beta, 1000 U/ml of TNF-alpha, and 1000 U/ml of IFN-gamma, as well as 1000 U/ml of porcine IFN-gamma. Cumulative insulin levels, insulin content, metabolic activity, and viability of these cells were examined. Additionally, nitric oxide production and the activity of antioxidant enzymes in these cells were also determined. RESULTS: TNF-alpha and the combination of the three human cytokines caused a transient increase in cumulative insulin levels. TNF-alpha alone enhanced insulin content on day 3. There was no effect of these human cytokines on mitochondrial function and viability. In contrast, porcine IFN-gamma inhibited fetal pig beta cell function and also caused their death. Adult pig islets are sensitive to the toxic effects of human TNF-alpha, IL-1beta, the combination of the three cytokines, and porcine IFN-gamma. The activity of the antioxidant enzymes catalase, glutathione peroxidase, and superoxide dismutase were significantly higher in fetal pig beta cells than in adult islets, implying that this may be the reason for the lack of adverse effects of the cytokines on the fetal beta cell. CONCLUSION: Fetal pig beta cells are resistant to the toxic effect of the human cytokines, TNF-alpha and IL-1beta, in vitro. This resistance suggests that fetal, but not adult beta cells, when transplanted into humans with type 1 diabetes may be protected from primary nonfunction and will be partially protected from autoimmune destruction. PMID- 11907417 TI - Porcine hematopoiesis on primate stroma in long-term cultures: enhanced growth with neutralizing tumor necrosis factor-alpha and tumor growth factor-beta antibodies. AB - BACKGROUND: Donor hematopoiesis is at a competitive disadvantage when bone marrow transplantation is across species barriers. This could present major limitations to xenogeneic stem cell transplantation as an approach to tolerance induction. An in vitro model of xenogeneic engraftment was established to identify inhibitors of porcine hematopoiesis in a primate environment. METHODS: Porcine bone marrow cells (BMC), in the presence or absence of primate CD34+ positive cells, were cultured for 4-6 weeks on primate stroma with porcine cytokines. Cellularity and growth of colony-forming cells were indicators of hematopoietic growth. Effects of soluble factors were determined by using Transwell inserts to separate porcine cells from stroma. Neutralizing antibodies for human transforming growth factor beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha) were added to cultures. RESULTS: Porcine hematopoiesis can be maintained in long-term cultures on primate stroma with pig cytokines. Adding BMC to the stroma below Transwell containing porcine cells dramatically inhibited porcine hematopoiesis, showing an inhibitory role for soluble factors. Neutralizing antibodies against TNF-alpha or TGF-beta caused a modest enhancement of porcine hematopoiesis; however, the combination of both led to a substantial increase. Inhibitory effects of these cytokines were confirmed by adding TNF-alpha and TGF-beta to porcine cultures. CONCLUSIONS: Porcine cells may be more sensitive to inhibitory effects of TNF alpha and TGF-beta than primate cells and are at a disadvantage when in a primate environment. Potential implications of this observation are discussed in the context of establishing specific immune tolerance via mixed chimerism to facilitate xenotransplantation. PMID- 11907418 TI - Different effects of tacrolimus and cyclosporine on renal hemodynamics and blood pressure in healthy subjects. AB - BACKGROUND: The side effects of cyclosporine, nephrotoxicity and hypertension, contribute to long-term renal graft failure and cardiovascular morbidity in graft recipients. It is not clear whether tacrolimus is as nephrotoxic and hypertensive as cyclosporine. Data on this subject are not consistent because of differences in dosage and duration of treatment and the presence of comorbidity in the studied patients. A comparison of both drugs with respect to renal hemodynamics and blood pressure has not been performed yet in healthy subjects. METHODS: We studied blood pressure, glomerular filtration rate, and effective renal plasma flow in eight healthy subjects at baseline and after 2 weeks administration of cyclosporine and tacrolimus, in randomized order. Trough levels of either drug were within the currently recommended therapeutical range of 100-200 ng/ml for cyclosporine and 5-15 ng/ml for tacrolimus. RESULTS: Tacrolimus did not influence renal hemodynamic parameters, in contrast to cyclosporine. During cyclosporine, glomerular filtration rate decreased from 98+/-9 ml/min/1.732 to 85+/-10 ml/min/1.732 (P<0.05), and ERPF decreased from 597+/-108 ml/min/1.732 to 438+/-84 ml/min/1.732 (P<0.01). Mean arterial blood pressure increased from 93+/-8 mmHg to 108+/-10 mmHg (P<0.05) during cyclosporine and remained unchanged during tacrolimus. CONCLUSIONS: We conclude that tacrolimus given during 2 weeks in the currently advised dosage has no unfavorable effects on renal hemodynamics and blood pressure in healthy individuals. The use of tacrolimus in organ transplant recipients may in the long-term lead to better renal function and less cardiovascular morbidity than the use of cyclosporine. PMID- 11907419 TI - Relationship between diabetes and obesity 9 to 18 years after hemipancreatectomy and transplantation in donors and recipients. AB - BACKGROUND: Healthy human volunteers occasionally elect to undergo surgical removal of the distal half of their pancreas for donation to a relative with type 1 diabetes. This provides the unusual opportunity to study segments of the same pancreas in two markedly different environments, i.e., the normal one of the donor and the unusual one of the ectopically transplanted recipient who is receiving immunosuppressant drugs that can diminish insulin secretion and cause insulin resistance. METHODS: We studied eight donor/recipient pairs 9 to 18 years after the original surgery to assess the hypothesis that beta-cell mass is the primary determinant of glucose homeostasis. We measured levels of fasting glucose and hemoglobin A1c, intravenous glucose disappearance rates, acute insulin and C peptide responses, and beta-cell secretory reserve. RESULTS: Comparisons of the mean data between the two groups revealed no significant differences in fasting plasma glucose, hemoglobin A1c, fasting insulin or C-peptide, acute insulin or C peptide responses to arginine and to glucose, or beta-cell secretory reserve. Eight patients were obese; this subgroup contained all patients who developed mild diabetes (four donors and two recipients). CONCLUSION: The within-pairs metabolic outcomes support the primacy of pancreatic mass in determining glucose homeostasis, but the discordancy within pairs for developing postoperative diabetes implicates variables, especially obesity, as important secondary determinants in the risk of developing diabetes in donors and recipients. Our data suggest that obesity should be a contraindication to donation of pancreatic segments and that donors should assiduously avoid becoming obese. PMID- 11907420 TI - Ketorolac-based analgesia improves outcomes for living kidney donors. AB - BACKGROUND: To reduce the morbidity of living kidney donors we introduced ketorolac-based analgesia for patients undergoing open donor nephrectomy in August 1999. There are no prior reports on the use of ketorolac for patients undergoing donor nephrectomy. METHODS: We conducted a retrospective review of all 198 consecutive patients who underwent open living donor nephrectomy between January 1998 and July 2000 at our institution. We compared patients who underwent surgery before and after introduction of ketorolac-based analgesia. RESULTS: The introduction of ketorolac-based analgesia resulted in a reduction in length of postoperative stay from 3.7+/-0.1 to 3.1+/-0.1 days (P<0.001). Patients who underwent surgery after ketorolac introduction required 58% less narcotics (P<0.001), recalled having less postoperative pain, and stopped taking pain medications sooner than patients before routine ketorolac use. Moreover, these patients drank 38% more liquids on the first postoperative day (P<0.001) and were able to resume a regular diet sooner than patients who were not managed with ketorolac. Patients who received ketorolac had a slightly lower creatinine clearance on the second postoperative day relative to patients who did not (66 vs. 72% of preoperative calculated creatinine clearance, P=0.05). However at a minimum of 3 months postoperatively, creatinine clearance did not differ between the two groups (70 vs. 73%, P=0.92). There were no differences in the rates of complications between the two groups. CONCLUSIONS: The use of ketorolac to control postoperative pain for patients undergoing open donor nephrectomy reduced morbidity and was not associated with any effect on long-term renal function or increased risk of complications. This is the first study to demonstrate the safety of using ketorolac at the time of donor nephrectomy. PMID- 11907421 TI - Progression of cardiomyopathy after liver transplantation in patients with familial amyloidotic polyneuropathy, Portuguese type. AB - BACKGROUND: Transthyretin amyloidosis is today an accepted indication for orthotopic liver transplantation (OLT). For several mutations progression of the cardiomyopathy has been observed after OLT. The aim of this study was to assess the course of cardiac involvement in Swedish familial amyloidotic polyneuropathy (FAP), Portuguese type, after OLT. By comparison of the echocardiographic findings before OLT with those obtained after, the course of the heart involvement was followed. METHODS: Twenty-three patients, who had undergone OLT and were examined with echocardiography 1-12 months before OLT, were available for the study. Twenty-one patients were examined 12-27 months after OLT, and 12 were re-examined 52-71 months after OLT. Two-dimensional and M-mode echocardiography were performed in accordance with the standards of the American Society of Echocardiography. RESULTS: A significantly increased septal and left ventricular posterior wall thickness and a significantly increased left atrial dimension was observed at the post-OLT examinations, indicating a progression of the amyloid heart disease. This increase of the cardiac involvement was neither correlated to waiting time for OLT or to pre-operative signs of cardiomyopathy. CONCLUSIONS: Even though the production of the amyloidogenic-mutated transthyretin is stopped by OLT, the cardiomyopathy may progress after the operation even for the Portuguese type of FAP. The increase of the septal and left ventricular posterior wall thickness after OLT is not restricted to patients with signs of left ventricular hypertrophy before the transplantation. The findings have important implications for the follow-up of FAP patients after OLT. PMID- 11907422 TI - Presence of autoantibody against ATTR Val30Met after sequential liver transplantation. AB - BACKGROUND: Recently, sequential liver transplantation has been performed with an explanted liver from a patient with familial amyloidotic polyneuropathy (FAP) because of the shortage of donors. However, metabolism of amyloidogenic transthyretin (ATTR), the pathogenic protein of FAP, has not been well studied in patients who have undergone sequential liver transplantation. The purpose of this study was to examine the changes in serum ATTR levels and to investigate the presence of an autoantibody in patients who underwent sequential liver transplantation with an explanted organ from a patient with heterozygotic FAP (FAP ATTR Val30Met). METHODS: This was a case study performed at the Kumamoto University School of Medicine, Kumamoto, Japan, and Kyoto University School of Medicine, Kyoto, Japan. Intervention occurred by sequential liver transplantation with an explanted FAP patient's liver. Levels of normal TTR and ATTR in the two patients who received the transplanted liver were analyzed by means of an enzyme linked immunosorbent assay (ELISA) and a matrix-assisted laser desorption/time-of flight mass spectrometry. In addition, the presence of an autoantibody against ATTR Val30Met was evaluated via ELISA using purified ATTR Val30Met from homozygotic FAP patients' sera. RESULTS: After the operation, the variant TTR levels were unexpectedly lower than levels of normal TTR in serum samples from patients with a transplanted liver from the FAP patient. An autoantibody against the variant TTR was detected on day 3 after the operation in the serum of those patients and continued to be present for at least 2 months after the operation. CONCLUSIONS: An autoantibody against the variant TTR may reduce the serum levels of variant TTR. Although the antibody may play a beneficial role in reducing the pathogenic protein, the long-term effect of the antibody must be investigated further. PMID- 11907424 TI - Correlation of chlamydia pneumoniae infection and severity of accelerated graft arteriosclerosis after cardiac transplantation. AB - BACKGROUND: Chlamydia pneumoniae has been associated with atherosclerosis, although its role in the process is not clearly defined. Heart transplant recipients are known to have high titers of antibodies to C. pneumoniae, and the organism has been recovered from the coronary arteries of both transplant recipients and donors. This study evaluated association between C. pneumoniae infection and accelerated graft arteriosclerosis (AGA), also known as cardiac allograft vasculopathy (CAV), after cardiac transplantation. METHODS: A case control study was performed with 54 heart transplant recipients at the Johns Hopkins Hospital. Severe cases had >50% luminal narrowing on cardiac catheterization, mild cases <50% narrowing, and controls were free of arteriosclerotic disease. Blood specimens were examined for C. pneumoniae serology and DNA detection by polymerase chain reaction (PCR) assays. RESULTS: For every twofold increase in geometric mean C. pneumoniae immunoglobulin (Ig)G titer, the odds ratio for severe AGA versus controls was 3.13 (P=0.03) and for mild AGA versus control patients was 1.61 (P=0.45). On Kaplan-Meier survival analysis there was a nonsignificant trend toward faster development of CAV in patients with higher C. pneumoniae antibody titers. Overall, 29% of heart transplant patients evaluated had evidence of circulating C. pneumoniae DNA by PCR, without a statistical difference between groups. CONCLUSIONS: C. pneumoniae IgG titer correlates with severity of allograft arteriosclerosis after cardiac transplantation. Circulating C. pneumoniae DNA is detectable by PCR in up to 30% of cardiac transplant recipients, but this does not correlate with severity of allograft vasculopathy. PMID- 11907423 TI - De novo hemolytic uremic syndrome after kidney transplantation in patients treated with cyclosporine-sirolimus combination. AB - OBJECTIVE: We sought to examine factors that predisposed 1.5% (10/672) of renal transplant recipients treated with a cyclosporine (CsA)/sirolimus (SRL)/steroid immunosuppressive regimen to develop hemolytic uremic syndrome (HUS). METHODS: Two cohorts of recipients were treated for 1-212 months (mean: 25.0+/-26.4, median: 18.1) with concentration-control CsA regimens based upon either area under the concentration-time curve (AUC; n=412 patients) or trough measurements (C0; n=260 patients). RESULTS: The only demographic feature more common to affected patients was an original glomerulopathic disease in 7 patients, 4 of whom had displayed IgA glomerulonephritis. All 10 affected patients showed a clinical picture of hemolysis with schistocytes, thrombocytopenia (nadir: 35,000+/-19,600 platelets/mm3), as well as elevated serum levels of lactate dehydrogenase (1697+/-1427 IU) and creatinine (Scr; 2.05+/-1.52 mg/dL prediagnosis to 5.13+/-2.43 mg/dL at diagnosis). Seven patients experienced adverse events concomitant with the bout of HUS, namely, acute rejection episodes prior to (n=2) or during (n=3), and 2 patients, infections (Herpes simplex and pancolitis). The mean values of daily steroid dose and the immunosuppressive drug C0 values were above the putative therapeutic targets: namely, CsA C0=294.9+/ 153.2 ng/ml versus 150+/-50 ng/ml and SRL C0=20.1+/-14.0 ng/ml versus 10+/-5 ng/ml, respectively. The therapeutic approach included discontinuation of CsA in 9/10, which was transient in 6/9; discontinuation of SRL in all 10, which was transient in 3, OKT3 for concurrent rejection in 3, and plasmapheresis in 5 patients. At 24 weeks postdiagnosis 9/10 patients have well-functioning kidneys with a mean Scr value of 1.6+/-0.59 mg/dL. One patient who underwent transplant nephrectomy subsequently succumbed due to a cluster of refractory thrombocytopenia, Aspergillus infection, and multiorgan failure. CONCLUSION: This initial experience suggests that a time-limited and reversible de novo HUS syndrome may be less frequent and milder among renal transplant recipients treated with SRL-based immunosuppression. PMID- 11907425 TI - Impaired volume regeneration of split livers with partial venous disruption: a latent problem in partial liver transplantation. AB - BACKGROUND: In living-donor and split-liver transplantations using a hemi-liver graft, it is practically impossible to maintain complete venous drainage in both the right and left livers, because the middle hepatic vein can be preserved only on the unilateral side. However, it is not clear whether partial venous disturbances affect postoperative liver volume regeneration. METHODS: Living donors who underwent left-sided hepatectomy preserving the middle hepatic vein (group A, n=40) or left hepatectomy with middle hepatic vein resection (group B, n=37) were reviewed. Volume regeneration of the remnant right paramedian (segments V + VIII) and lateral (segments VI + VII) sectors and overall liver volume was assessed at 3 postoperative months by computed tomography. RESULTS: In group A, both sectors showed a proportional increase by 21.7% (P=0.991), whereas in group B the rate of increase of the right paramedian sector was less than that of the right lateral sector (13.3% vs. 36.5%, P<0.001). Comparisons of rate of increase for each sector between the groups indicated that interruption of the middle hepatic venous drainage impaired enlargement of the right paramedian sector and induced a compensatory hypertrophy of the right lateral sector. Overall liver mass restoration rate in group B was inferior to that in group A (78.9% vs. 85.0%, P=0.001). CONCLUSIONS: Split livers with partial outflow disturbances are associated with latent disadvantages in postoperative liver volume regeneration even if venous congestion is not evident. These results suggest a problem of regenerative capacity of right liver grafts. PMID- 11907426 TI - Cimetidine improves prediction of the glomerular filtration rate by the Cockcroft Gault formula in renal transplant recipients. AB - BACKGROUND: The glomerular filtration rate (GFR) can be predicted from plasma creatinine, age, gender, and body weight, using the formula of Cockcroft and Gault. Cimetidine improved the accuracy of GFR prediction in renal disease and also in diabetes mellitus type 2, due to inhibition of tubular creatinine secretion. We compared the accuracy and precision of GFR prediction from the Cockcroft-Gault formula without cimetidine (CG), with cimetidine (CGcim) and from the creatinine clearance without cimetidine in renal transplant recipients. METHODS: CG and CGcim were calculated from plasma creatinine before and after 2400 mg of oral cimetidine during the 24 hr preceding the GFR measurement. The endogenous creatinine clearance was measured in 24 outpatients from a 24-hr urine collection (Ccr24) before cimetidine. GFR was measured as the urinary clearance of continuously infused 125I-iothalamate. Creatinine was determined with an automated enzymatic assay in plasma and with an alkaline picrate assay in urine. RESULTS: GFR was 47.8+/-16.8 ml/min/1.73 m2 (mean+/-SD), Ccr24 was 71.8+/-23.1 ml/min/1.73 m2, CG was 62.2+/-15.2 ml/min/1.73 m2, and CGcim was 52.8+/-14.9 ml/min/1.73 m2. Ccr24 overestimated GFR in every patient by an average of 23.8 ml/min/1.73 m2 and CG by an average of 14.3 ml/min/1.73 m2, whereas CGcim overestimated GFR significantly less by an average 4.9 ml/min/1.73 m2 (P<0.001). The precision of CGcim was significantly better than that of Ccr24: the SD of the difference from GFR was 9.0 ml/min/1.73 m2 for CGcim and 14.5 ml/min/1.73 m2 for Ccr24 (P<0.05). CONCLUSION: CGcim is useful for GFR prediction in outpatient renal transplant recipients and has a far better accuracy and precision than Ccr24 and also a better accuracy than CG. We propose a strategy after kidney transplantation of one GFR measurement at baseline and follow-up with CGcim. PMID- 11907427 TI - A long-term comparison of tacrolimus (FK506) and cyclosporine in kidney transplantation: evidence for improved allograft survival at five years. AB - BACKGROUND: The 1-year results of the Phase III U.S. Multicenter Trial comparing tacrolimus (FK506)- and cyclosporine (CsA)-based immunosuppressive therapy in kidney transplantation revealed a significant reduction in the incidence and severity of acute rejection episodes among patients maintained on tacrolimus. The present report at 5 years of follow-up focuses on the long-term impact of tacrolimus treatment on kidney allograft outcome. METHODS: The study protocol permitted crossover of patients to the alternate treatment arm under stringent conditions. The effect of crossover on graft survival was analyzed. Cardiovascular risk factors and serious adverse events were also monitored over 5 years. RESULTS: Intent-to-treat analysis revealed equivalent patient and graft survival between treatment arms at 5 years of follow-up (79.1% vs. 81.4%; P=0.472 and 64.3% vs. 61.6%; P=0.558 among tacrolimus and CsA-treated patients, respectively). However, the rate of crossover was significantly higher among patients randomized to receive CsA-based therapy (27.5% vs. 9.3%; P<0.001). The incidence of treatment failure (43.8% vs. 56.3%; P=0.008) was significantly lower among tacrolimus-treated patients. Graft survival was significantly improved in the tacrolimus treatment arm when crossover due to rejection was counted as graft failure (63.8% vs. 53.8%; P=0.014). Tacrolimus therapy was also associated with a significantly reduced requirement for medications to control hypertension and hyperlipidemia. There was a substantial rate of reversal of tacrolimus-associated insulin dependence. CONCLUSION: Tacrolimus-based therapy resulted in significantly reduced risk of graft failure, without an increase in the incidence of adverse events associated with long-term immunosuppression. PMID- 11907428 TI - Angiotensin converting enzyme inhibition in chronic allograft nephropathy. AB - BACKGROUND: Although angiotensin-converting enzyme inhibition has been shown to slow progression of chronic allograft nephropathy in animal models, no studies have examined its efficacy in humans. METHODS: We retrospectively analyzed 63 patients with biopsy-proven chronic allograft nephropathy who had > or =6 months dialysis-free follow-up at our institution. A total of 32 patients treated for > or =6 consecutive months with angiotensin-converting enzyme inhibition and/or angiotensin-receptor blocker (ARB) therapy (group 1) were compared with 31 patients not on these agents (group 2). RESULTS: Except for a higher incidence of hypertension (100 vs. 78%, P=0.005) in group 1, there were no significant differences in baseline clinical characteristics at time of biopsy. With a mean follow-up time of 27 months in both groups, 6 of 32 (19%) group 1 patients vs. 12 of 31 (39%) group 2 reached the primary endpoint of > or =50% increase in serum creatinine (P=0.10). Mean time to primary endpoint was 46.6 months in group 1 vs. 32.7 months in group 2 (P=0.07). Three of 32 (9%) of group 1 patients vs. 8 of 31 (26%) of group 2 returned to dialysis during this time (P=0.11). Significantly fewer patients in group 1 reached the combined secondary endpoint of allograft failure or death (9.4 vs. 35.5%, P=0.01); in addition, group 1 had a longer mean time to this endpoint (51.2 vs. 37.6 months, P=0.03). On multivariate analysis, the only predictor of progression to primary endpoint was a high baseline serum creatinine (P=0.02). No significant differences in hyperkalemia or anemia were found between the two groups. CONCLUSIONS: Angiotensin-converting enzyme inhibition/angiotensin-receptor blocker therapy is well tolerated in renal allograft recipients with chronic allograft nephropathy. It is associated with a trend of slowing renal insufficiency as well as a significant survival benefit in the combined endpoint of allograft failure or death. PMID- 11907429 TI - Rolling adhesion of human NK cells to porcine endothelial cells mainly relies on CD49d-CD106 interactions. AB - BACKGROUND: Acute vascular rejection in pig-to-primate xenotransplantation involves recognition and damage of porcine (po) endothelial cells (EC) by human (hu) leukocytes, probably including natural killer (NK) cells. To study such interactions we analyzed rolling and static adhesion of hu NK cells to po EC. METHODS: The effects of blocking hu and po adhesion molecules on the adhesion hu NK cells to po EC monolayers was analyzed under shear stress (10 min, 37 degrees C, 0.7 dynes/cm2) or under static conditions (10 min, 37 degrees C). All used cell populations were phenotypically characterized by flow cytometry. RESULTS: Blocking of CD106 on po EC or its ligand CD49d on hu NK cells decreased rolling adhesion of both fresh and activated hu NK cells by more than 75%. Masking of CD62L on fresh but not activated hu NK resulted in a 44% decrease in rolling adhesion, in line with the diminished cell surface expression of CD62L upon activation. Antibodies to CD31, CD54, CD62E, and CD62P on EC or CD11a, CD18, and CD162 on NK cells had only minor effects on rolling adhesion. The adhesion of the FcgammaRIII- hu NK cell line NK92 to po EC was inhibited by 95% after masking po CD106 whereas antibodies to po CD31, CD54, CD62E, or CD62P had no effect, thereby excluding effects of Fc-receptor-dependent binding of hu NK cells to po EC. Static adhesion of activated NK cells was reduced by approximately 60% by blocking either CD49d or CD106, by 47% by blocking CD11a, and by 82% upon simultaneous blocking of CD11a and CD49d. CONCLUSIONS: Interactions between hu CD49d and po CD106 are crucial for both rolling and firm adhesion of hu NK cells to po EC and thus represent attractive targets for specific therapeutic interventions to prevent NK cell-mediated responses against po xenografts. PMID- 11907430 TI - Factors influencing the patterns of T lymphocyte allorecognition. AB - BACKGROUND: Strong alloreactive T cell responses are a menace in transplantation surgery and their menagement requires understanding the basis of alloreactivity. Alloantigen recognition can be peptide independent, peptide specific, or peptide dependent. The mechanisms influencing each recognition pattern are largely unknown. METHODS: Peptide dependence was examined in vitro by adding peptides to antigen processing-deficient cell line used as target in cytotoxic T cell assays. Responses to major histocompatibility complex (MHC) alleles most homologous to self were recently shown to be more peptide dependent than to those with lesser homology to self. Hence, peptide reactivity in vivo was estimated based on relative strengths of alloreactive responses to more homologous and less homologous MHC alleles. RESULTS: Alloreactive CD8+ TCR repertoire in beta2 microglobulin-deficient mice is preferentially peptide independent. The peptide specific component is acquired as a function of wild-type thymic epithelium grafting. Irrespective of the presence of the peptide-specific component, in vivo alloantigenic priming was associated with a greater sensitivity to the MHC structure than was in vitro priming. CONCLUSIONS: Thymic positive selection and the mode of alloreactivity induction are the major independent factors determining the patterns of alloantigen recognition. PMID- 11907431 TI - T cells from newborn humans are fully capable of developing into cytotoxic T lymphocyte effector cells in adoptive hosts. AB - BACKGROUND: The finding of reduced incidence of graft-versus-host disease (GVHD) associated with cord blood transplantation, compared to unrelated allogeneic bone marrow, could be related to the lower number of T cells infused in the cord blood (CB) inoculum or it might represent an intrinsic property of CB T cells. We investigated the in vivo function of human cord blood mononuclear cells (CBMC) after their adoptive transfer into lethally irradiated BALB/c radioprotected with severe combined immunodeficiency (SCID) mouse bone marrow. METHODS: The ability of human CBMC to engraft and produce antigen-specific alloreactive cytotoxic T lymphocytes (CTLs) and antibodies was determined by FACS, 51Chromium-release assay, and ELISA. RESULTS: Recipients of human CBMC showed engraftment of high levels of both CD14+ and CD3+ cells. Human cells recovered from the peritoneal cavity of chimeric mice, 1 week after immunization with irradiated allogeneic cells, showed adult-level human CTL response against the immunizing cells, without further stimulation in vitro. In contrast, immunization with the tetanus (TT) antigen did not lead to the generation of anti-TT immunoglobulins (Ig) in the cord blood chimera. Furthermore, whereas the addition of purified adult T cells to the cord blood inoculum resulted in the enhancement of human Ig production of both IgG and IgM subclasses, it could not induce antigen-specific antibodies after immunization. CONCLUSION: This report demonstrates in mice for the first time the generation of classical human alloreactive CTLs, derived from cord blood cells. The alloreactivity exhibited by the cord blood mononuclear cells is not different from that displayed by cells originating from adult blood. Any reduction in the observed GVHD associated with cord blood transplants might therefore represent a quantitative difference in the total number of T cells infused. PMID- 11907432 TI - Proposed guidelines for re-evaluation of patients on the waiting list for renal cadaver transplantation. AB - Transplant candidates are extensively evaluated before being wait-listed for cadaver transplantation. Yet many wait a number of years before being transplanted. We propose guidelines for regular cardiac re-evaluation for patients on the waiting list. PMID- 11907433 TI - Limited efficacy of lamivudine against hepatitis B virus infection in allogeneic hematopoietic stem cell transplant recipients. AB - BACKGROUND: Reactivation of chronic hepatitis B virus (HBV) infection is a major complication when HBV carriers receive immunosuppressive therapy. Recipients of allogeneic hematopoietic stem cell transplantation (HSCT) carry the highest risk of fatal HBV disease (up to 12%). METHODS: In an attempt to identify a suitable procedure for the prevention and management of HBV reactivation, the administration of lamivudine over the course was tested in two patients. RESULTS: Generally, the patients transplant courses were successfully managed despite their difficult clinical situations: a high HBV load before transplant in one patient and intense steroid therapy for complicated acute graft-versus-host disease (GVHD) in the other patient. However, one patient showed a reactivation of HBV after discontinuing lamivudine and the other showed persistently high DNA polymerase activity despite prolonged administration of lamivudine. CONCLUSIONS: We concluded that lamivudine could have a place in the management of patients who suffer from chronic HBV infection and who are undergoing allogeneic HSCT. However, the efficacy of lamivudine seemed to be limited compared with other settings, including solid organ transplantation and autologous HSCT. PMID- 11907434 TI - Autoimmune and pregnancy complications in the daughter of a kidney transplant patient. AB - BACKGROUND: Immunosuppressive agents taken by pregnant organ transplant recipients readily cross the placenta during development of the fetal immune system. There are few data on the long-term implications for the progeny, but evidence from animal studies suggest that second and third generations of organ transplant patients may be at risk for autoimmune disorders and reproductive problems. METHODS: We present the 23-year-old daughter of a renal allograft recipient exposed to azathioprine 75 mg/day and prednisone 5 mg/day throughout her mother's pregnancy. RESULTS: During the daughter's first pregnancy, she developed multiple autoantibodies, Raynaud's phenomenon, and fetal death occurred at 20 weeks gestation. The second pregnancy was complicated by systemic lupus erythematosus, preeclampsia, and the birth of a preterm male infant. CONCLUSIONS: It is uncertain whether the autoimmune manifestations and obstetric complications in this patient were related to fetal exposure to immunosuppressive drugs. Nevertheless, further studies on the health and pregnancies of adult offspring of transplant patients are warranted. PMID- 11907435 TI - Sensitization of renal transplant candidates by cryopreserved cadaveric venous or arterial allografts. AB - BACKGROUND: Cryopreserved cadaveric venous or arterial allografts are used in ESRD patients as an alternative to synthetic grafts for hemodialysis access. We evaluated the effect of these allografts on the PRA against HLA class I and II antigens in 11 ESRD patients awaiting a kidney transplant. METHODS: Flow Cytometry using purified antigen coated beads (Flow PRA Beads) was used to determine PRA against HLA class I and II antigens. RESULTS: Patients with no antibody prior to allograft implantation were all positive for HLA class I and II antibodies after implantation. Patients with anti-HLA class I and II antibodies prior to allografting, developed higher antibody titers. Of the 11 patients that received cryopreserved cadaveric allografts no deaths were reported. Two grafts were removed due to thrombosis consistent with rejection. CONCLUSION: Recipients of cadaveric venous or arterial allografts elicit an HLA antibody response attesting to the antigenicity of cryopreserved cadaveric allografts. PMID- 11907436 TI - Seroconversion after the addition of famciclovir therapy in a child with hepatitis B virus infection after liver transplantation who developed lamivudine resistance. AB - There is very little information about hepatitis B virus (HBV) infection in children after liver transplantation. This is the first report of the addition of famciclovir in a child who developed lamivudine resistance.A 5-year-old boy who was serum HBsAg-negative and was not vaccinated against HBV underwent living related liver transplantation for fulminant hepatitis A. The donor was his mother, who was serum HBcAb-positive. No immunoprophylaxis was administered. HBV infection developed after 18 months and was treated with 3 mg/kg daily of lamivudine. Serum alanine aminotransferase normalized and HBV DNA load decreased significantly. Sixteen months later, lamivudine resistance developed; a mutation (M552I) was confirmed by sequencing through the YMDD locus of the HBV polymerase gene. The addition of 750 mg daily of famciclovir led to seroconversion and the disappearance of serum HBV DNA. Lamivudine in combination with famciclovir might be a therapeutic option for HBV reinfection after liver transplantation, also in children. Suppression of viral replication to undetectable values is possible even in the lamivudine-resistant mutant. PMID- 11907437 TI - Real-time sequence-specific primer polymerase chain reaction amplification of HLA class II alleles: a novel approach to analyze microchimerism. AB - The careful assessment of microchimerism is essential to investigate the effects of donor bone marrow-derived cells in transplantation. We have developed a protocol to assess microchimerism based on the HLA mismatch between the recipient and the donor. Our approach combines real-time polymerase chain reaction (PCR) with sequence-specific primer PCR (SSP-PCR) to selectively amplify and measure the abundance of donor HLA alleles in DNA samples extracted from the recipient after transplant. To optimize and validate the reliability of this method at different levels of microchimerism, we tested serial dilutions of donor DNA into recipient DNA. We demonstrate that donor alleles can be readily detected and reliably measured at concentrations as low as 0.1%. This method is simple and rapid and could find practical application in the assessment of microchimerism in patients receiving organ or cellular transplants in conjunction with donor bone marrow cells infusion. PMID- 11907438 TI - Vascularized thymic lobe transplantation in miniature swine: I. Vascularized thymic lobe allografts support thymopoiesis. AB - BACKGROUND: Vascularized thymokidney transplants have previously been shown to induce tolerance across major histocompatibility complex barriers. The ability to perform vascularized thymic lobe transplantation could permit such tolerance to be induced with any cotransplanted solid organ or tissue. For this reason, we have developed a technique for vascularized thymic lobe transplantation in miniature swine. METHODS: Thymic vessels (n=2) were anastomosed to the carotid artery and the external jugular vein of naive minor-mismatched recipients treated with a 12-day course of cyclosporine A (10 mg/kg/day). Graft survival and thymopoiesis were assessed by histology, immunohistochemistry, and fluorescence activated cell sorting. Allele-specific antibodies 74-12-4 and pig allelic antigen (PAA) were used to distinguish donor and recipient cells. RESULTS: Allografts showed intact cortical and medullary structure posttransplantation, without evidence of rejection or ischemia. Recipient thymocytes repopulated the donor cortical thymus by POD30 and increased in the cortex and medulla by POD60. CONCLUSIONS: Our study demonstrates the technical feasibility of vascularized thymic lobe transplantation and the support of thymopoiesis by such transplants in a large animal model. This technique may offer a novel strategy to induce transplant tolerance across allogeneic and xenogeneic barriers, and to support long-term thymopoiesis in immunodeficient hosts. PMID- 11907439 TI - Composite tissue allotransplantation. PMID- 11907440 TI - There's no alternative to taxation. PMID- 11907441 TI - Venous haemodynamics and the effects of compression stockings. AB - Over the years, graduated compression stockings exerting varying ankle pressures have been used with differing degrees of effectiveness in the prevention of deep venous thrombosis and ulcer recurrence, and the treatment of venous ulceration (Stemmer et al, 1980; Partsch and Horakova, 1994; Veraart et al, 1997). Whether in the hospital or community setting, nurses often have the responsibility of measuring the limb and fitting the compression stocking on the patient and it is he/she who often influences the patient on the type of stocking and level of compression required. Understanding the influence of graduated compression on the venous haemodynamics of the lower limb and having a good working knowledge of the claims of the manufacturers as to expected levels of compression from each garment will aid the nurse and patient in decision making. An awareness of the hazards of inappropriate use of compression therapy should ensure that the nurse is a safe practitioner. PMID- 11907442 TI - Early detection of type 2 diabetes: the role of the community nurse. AB - Recent changes to the diagnostic criteria for type 2 diabetes have raised concerns that if steps are not taken to address relevant issues, the number of diagnosed sufferers will escalate in future years, with a similar increase in the prevalence of associated complications. Research suggests that for many patients, complications are evident at the time of diagnosis, and may have been present for several years before a diagnosis was made. Late presentation is the cause of considerable morbidity and mortality among these patients, largely because undiagnosed complications are often irreversible by the time the underlying diabetes is discovered. The current emphasis on public health augments the need for community nurses to recognize warning signs, particularly in patients who are at an increased risk of developing diabetes. This article aims to raise awareness of early signs and symptoms, and gives a brief insight into some of the associated health problems. If community nurses are to play a significant role in reducing the potentially devastating effects of this disease, they will need to be involved tin educating patients and implementing screening and intervention programmes. PMID- 11907443 TI - The management and prevention of incontinence dermatitis. AB - Incontinence dermatitis (ID) is rising in incidence along with the increasing elderly population. Its management must therefore take on a greater level of importance. The focus of care must be on finding the causes of incontinence; in many cases it can be cured or the symptoms greatly improved and the development of ID prevented. Where it does occur, effective intervention is necessary. However, a survey into the knowledge base of nurses in caring for skin, carried out by the Royal College of Nursing Continence Care Forum in 1995, showed that care was based on customs and practice rather than evidence. This often causes incorrect treatment to be given, resulting in extra cost for the National Health Service (NHS). This article examines the aetiology and presentation of ID, and supports the need for greater dissemination of guidelines for incontinence professionals and the need for evidence-based literature. PMID- 11907444 TI - Diagnosis and management of fungal infections of the skin. AB - Fungal infections of the skin are very common in the general population, and will be among the conditions most frequently encountered by nurses in primary care. In this article the types of fungal infections that the nurse will meet are described and the signs and symptoms outlined. The diagnosis of each fungal infection will be discussed as well as the treatment that is available to the nurse in these instances. The importance of preventing re-infection and transmission of the fungi is highlighted, and ways of prevention are also discussed. PMID- 11907445 TI - Record keeping and nurse prescribing: an issue of concern? AB - As options are explored by the NHS Executive to extend nurse prescribing in terms of expanding the Nurse Prescribers' Formulary and widening the groups of nurses eligible to train to prescribe, issues surrounding record keeping in the community need to be revisited in order to safeguard professional integrity. Prescribing nurses in the community face barriers to accurate and adequate record keeping when access to all patients' records is not permitted and/or time constraints and extra travelling impede the process, potentially threatening the safety of the patient. Until information technology provides the answer to safe and effective use of shared patient records, nurses who prescribe any treatments need to ensure they share full access to all patients' notes. PMID- 11907446 TI - MMR vaccination: the case for evidence-based practice. PMID- 11907447 TI - Nurse referrals of children to social services. AB - This literature review examines current referral systems and processes used by community nurses when making referrals of children in need and children in need of protection to social services departments. The problems surrounding definitions of need, children in need and children in need of protection are considered. The ambiguity of the language used and the difficulties encountered by community nursing staff in trying to identify criteria and thresholds for referral are highlighted. The review focuses on research exploring the assessment and referral processes used by community nurse, in particular by health visitors, and draws attention to the lack of research concerning these areas. Government recognition of the problems encountered by agencies working in this field is acknowledged in recent Department of Health publications. PMID- 11907448 TI - What constitutes optimal nutrition? PMID- 11907450 TI - An enduring monument to a crude injustice. PMID- 11907449 TI - Agency nurses must not be made scapegoats. PMID- 11907451 TI - Patient who was left on a commode for over eight hours. PMID- 11907452 TI - Cardiac rehabilitation: the needs of South Asian cardiac patients. AB - There is an indication that South Asian people in the UK experience greater delays than white British populations in obtaining appropriate treatment and intervention despite experiencing higher levels of coronary heart disease (Chaturvedi et al, 1997). Evidence suggests that access to and uptake of UK cardiac rehabilitation services is disproportionately low in South Asian populations (NHS Centre for Reviews and Dissemination, 1998). This article examines the results of an audit of cardiac rehabilitation among cardiac patients of South Asian origin who were admitted to a large city teaching hospital in Sheffield. The results are discussed in the light of current concerns about the adequacy of communication with non-English speaking NHS patients. The implications for access to services and clinical practice are considered. PMID- 11907453 TI - The management of ischaemic and periampuation pain. AB - This article describes the development and pilot application of a management tool designed to support healthcare practitioners in the management of ischaemic and periamputation pain. The ischaemic management periamputation care (IMPAC) guidelines have been piloted in the authors' clinical area and early results suggest that there are positive benefits accruing to patients and ward staff. PMID- 11907454 TI - Legal aspects of consent 12: organ donation after death. PMID- 11907455 TI - Central venous catheters: infection and patient susceptibility. AB - Infection is a major cause of morbidity in haemodialysis patients, of which between 25% and 50% of infections are related to vascular access, most commonly central venous catheters (CVCs) (Kessler et al, 1993). The morbidity associated with CVC infection provides a focus for research investigation, as there is limited knowledge available in relation to physiological and psychological parameters that may increase the probability of haemodialysis patients acquiring a CVC-related infection. This study explores these issues and discusses various factors that influence infection. It then describes how organisms which live on and inside our bodies can become parasitic, thus causing an infection. The precautions taken by healthcare professionals to reduce colonization and infection are enumerated, and the value of considering patients' physiological and psychological parameters alongside the more traditional extrinsic infection control approaches is discussed. PMID- 11907456 TI - Treatment of a wound infection in a patient with mantle cell lymphoma. AB - This case study examines the impact of a severe wound infection on a patient undergoing chemotherapy for the treatment of mantle cell lymphoma. The study illustrates how life threatening an infection can become in a patient whose body is compromised as a consequence of both disease and chemotherapy treatment. A number of specialist products were required in order to overcome the infection, debride and heal the wound. These included larval therapy, vacuum-assisted closure and Leptospermum honey. While this case study focuses on the successful outcome of one particular patient it aims to raise awareness of the role of specialist products, both old and new, in improving healing rates of complex wounds. PMID- 11907457 TI - Outpatient nurses: from handmaiden to autonomous practitioner. AB - Little seems to have been written about health promotion in outpatients' departments. This article argues that outpatient nurses have the potential to play a significant part in promoting people's health. Outpatient nurses may be the first hospital nurses that an individual meets. What people observe and how they are treated may make a difference to both their experience of hospital care and their health. Outpatient nurses may have been perceived as the doctors' handmaidens of yesterday, but today they are evolving into an effective force for change. For example, evaluation of nurse-led outpatients' clinics is showing that they have the potential to complement doctor-led clinics and improve the healthcare experience of patients. It is argued that the health-promoting role of the outpatient nurse can be enhanced by a department that is a health-promoting environment. PMID- 11907458 TI - Esprit HR mattress cover in pressure ulcer prevention. AB - Modern mattresses provide soft dense foam, which permits the redistribution of pressure on the patient over a wider area - away from bony prominences where pressure ulcers usually occur. The material used in producing multistretch covers for the new mattresses had a tendency to delaminate as a result of a combination of heat, moisture and inappropriate cleansing techniques causing the water barriers to fall. In partnership with York Health NHS Trust and the material manufacturer, STM Healthcare produced a mattress cover (Esprit HR) which was able to withstand higher pressure from heat and moisture and greatly extended the life expectancy of the Esprit HR mattress. PMID- 11907459 TI - Is nursing care losing its holistic focus? PMID- 11907460 TI - The promise of new technologies and therapies. PMID- 11907461 TI - The new melanoma staging system. AB - BACKGROUND: Classification schemas for cancers are useful for predicting overall survival and selecting patients for treatment. Historically, the most important factors in determining prognosis in patients with melanoma have been tumor thickness and lymph node status. Sentinel lymph node mapping defines a subset of patients with microscopic metastatic disease can be identified, offering greater accuracy in staging. METHODS: The authors reviewed studies evaluating the prognostic factors that are significant in predicting survival in patients with melanoma. The newly revised American Joint Committee on Cancer (AJCC) staging system for melanoma is compared with the 1997 AJCC staging system currently in use. RESULTS: The changes in the new AJCC melanoma staging system reflect the new prognostic factors that have been found to be important in predicting survival. These include primary tumor thickness (tumor depth in millimeters is more predictive than the level of invasion) and ulceration, number of metastatic lymph nodes, micrometastatic disease based on the sentinel lymph node biopsy technique or elective node dissection, the site(s) of distant metastatic disease and serum LDH levels. CONCLUSIONS: Major revisions have been made to form a new AJCC staging system for melanoma, which will become official in 2002. This system will provide more accurate and precise information regarding patient prognosis. Validation studies are needed to confirm the accuracy of this revised staging system. PMID- 11907462 TI - An update on adjuvant interferon for melanoma. AB - BACKGROUND: Despite advances in the staging and surgical therapy of melanoma, patients with high-risk resected melanoma still have 5-year recurrence rates of 55% to 80% and 5-year survival rates as low as 25% to 70%. Effective adjuvant therapy is needed for this patient population. METHODS: The authors review the literature regarding the use of interferon for the adjuvant therapy of resected melanoma. RESULTS: Low-dose adjuvant interferon regimens have not affected overall survival and have had an inconsistent effect on disease-free survival across different stage groupings. High-dose adjuvant interferon improved disease- free and overall survival in the E1684 and Intergroup E1694 trials. High-dose interferon regimens cause significant morbidity, but quality-adjusted years of life are greater with this therapy. CONCLUSIONS: Adjuvant high-dose interferon should be considered standard therapy for all high-risk melanoma patients expected to be able to tolerate the interferon and treated off protocol. In addition, this regimen should serve as the active control in future trials of alternative adjuvant therapies for these patients. PMID- 11907463 TI - Immunotherapy for melanoma. AB - BACKGROUND: Immunotherapy for cancers is based on the principle that the host's immune system is capable of generating immune responses against tumor cells. Currently available treatments for melanoma patients are limited by poor response rates. Interferon-a has been approved for adjuvant treatment of stage III melanoma with improved survival. New and more innovative approaches with improved efficacy are needed. METHODS: We reviewed the various new approaches and strategies for immunotherapy for the treatment of melanoma. RESULTS: Immunotherapy for melanoma includes a number of different strategies with vaccines utilizing whole cell tumors, peptides, cytokine-mediated dendritic cells, DNA and RNA, and antibodies. CONCLUSIONS: A variety of approaches can be used to enhance immune reactivity in patients with melanoma. Preclinical studies and initial clinical trials have shown promising results. Additional clinical trials are currently ongoing to evaluate the clinical efficacy and the associated toxicities of these novel treatment strategies. PMID- 11907464 TI - Metastatic melanoma: chemotherapy to biochemotherapy. AB - BACKGROUND: Single-agent or combination chemotherapy regimens have not impacted the short median survival of patients with metastatic melanoma, and complete or durable responses are rare. Biologic response modifiers (interferon and interleukin-2) have produced durable remissions in a small cohort of patients, and phase II trials of biochemotherapy suggest more benefit. METHODS: The authors retrospectively reviewed the status of the current treatments of metastatic melanoma focusing on biochemotherapy. RESULTS: Regimens include both sequential and concurrent approaches for inpatient and outpatient treatment settings. Overall response rates in phase II trials are 40% to 60% with complete responses of 10% to 20% and median survivals in the 11- to 12-month range. Modifications of concurrent biochemotherapy regimens have maintained efficacy and reduced toxicity. Small phase III trials suggest a survival advantage of biochemotherapy (P=.05). CONCLUSIONS: Biochemotherapy remains a promising new treatment for metastatic melanoma. A large Intergroup trial E3695 comparing concurrent biochemotherapy to combination chemotherapy alone is powered to answer important survival questions. PMID- 11907466 TI - The promise of microarray technology in melanoma care. AB - BACKGROUND: Genetic aberration is responsible for the development of neoplastic potential in a number of malignancies. These DNA alterations result in significant changes in gene expression that may now be measured and catalogued. The microarray technique screens and identifies expressed genes that may be responsible for tumorigenesis. METHODS: The authors review the application of the microarray technique in malignant melanoma. RESULTS: Candidate melanoma suppressor genes have been identified in melanoma cell lines using this technique. Furthermore, molecular classification using gene expression profiling may improve the accuracy of the staging system for determining prognosis. CONCLUSIONS: The microarray technique is in its initial development for clinical application in a variety of tumor models. Melanoma is an ideal system to study the genetic changes associated with the stepwise progression of malignancy. It may be possible to efficiently screen the entire human genome to identify the particular aberrations in gene expression responsible for tumorigenesis in melanoma. PMID- 11907465 TI - Advances in gene therapy for malignant melanoma. AB - BACKGROUND: The recent developments in the field of gene transfer have advanced the use of gene therapy as a novel strategy against a variety of human malignancies. Due to its unique set of characteristics, melanoma represents a suitable target for the clinical translation of the different gene transfer approaches recently developed. The goal of gene therapy targeted to melanoma cells is to introduce "suicide" genes, to transfer tumor suppressor genes, to inactivate aberrant oncogene expression, or to introduce genes encoding immunologically relevant molecules. Gene therapy targeted to the host's immune cells has been developed as an additional strategy to redirect immune responses against melanoma. METHODS: The authors reviewed the published gene transfer studies in experimental models, as well as the results of gene therapy clinical trials for patients with melanoma. RESULTS: Clinical trials have shown the feasibility and safety of gene therapy against malignant melanoma. Although no major successes have been reported, the positive results observed in some patients support the potential for gene therapy in the management of this disease. CONCLUSIONS: Gene therapy of melanoma using current gene transfer approaches is feasible and safe. Better vector technology as well as increased understanding of the "bystander effect" triggered by gene transfer approaches would provide the tools to validate gene therapy as an effective modality of treatment for malignant melanoma. PMID- 11907467 TI - Cancer economics: on variations in the costs of treating cancer. PMID- 11907468 TI - Hepatic surgery for metastatic gastrointestinal neuroendocrine tumors. PMID- 11907469 TI - Imaging in oncology: radiologic abnormalities in the legs. PMID- 11907471 TI - Correcting esotropia-time is of the essence! PMID- 11907472 TI - Outcome study of stereopsis in relation to duration of misalignment in congenital esotropia. AB - PURPOSE: A recent outcome study of Randot stereopsis in congenital esotropia reported early surgical alignment is associated with a higher percentage of patients with stereopsis and a higher quality of stereopsis because early surgery minimized the duration of misalignment. We compared the Titmus stereopsis with the duration of misalignment and the age of alignment for a group of patients who were surgically aligned by 24 months of age. METHODS: The data previously obtained in a study of 90 patients aligned by 2 years of age, examined in a masked, independent evaluation for binocularity was analyzed. The age of alignment and duration of misalignment was correlated with the percentage of patients with stereopsis and quality of the stereopsis result as determined by the Titmus vectograph overlay. RESULTS: Patients aligned by 6 or 12 months of age or within 6 or 12 months of duration of misalignment did not differ in percentage with stereopsis. However, patients aligned after 12 months of age did show a decrease percentage with stereopsis (P <.05, power 0.8). The quality of the stereopsis that was established was similar for those aligned by 6 or 12 months (P >.05, power 0.8). The quality of stereopsis, however, was decreased for patients with duration of misalignment greater than 12 months (P <.001, power 0.8). CONCLUSION: Alignment within 1 year of age or within 12 months of misalignment favorably affects the percentage of patients who develop stereopsis in the treatment of congenital esotropia. The quality of the stereopsis result is affected by the duration of the misalignment, rather, than the age of alignment per se. PMID- 11907473 TI - Complications in the first year following cataract surgery with and without IOL in infants and older children. AB - PURPOSE: The optimal role of intraocular lenses (IOLs) in infants remains a controversial topic for many reasons, including concerns about significant complications occurring in young rapidly developing eyes. METHODS: To assess the number and type of significant complications requiring further intervention occurring in the first postoperative year, we reviewed the records of 15 eyes of 13 infants undergoing lensectomy with posterior chamber IOL and pars plana vitrectomy (PPV)/capsulectomy under 6 months (group A) of age as part of an ongoing prospective study of IOL use in infants. This group was compared with a group of 16 children age 10 months to 5 years undergoing an identical procedure (group B) and a group of 33 infants less than 6 months of age undergoing lensectomy/vitrectomy without IOL (group C). RESULTS: Thirteen of 15 eyes in group A required additional surgery in the first postoperative year. Twelve of the 15 eyes (80%) developed secondary opacification across the visual axis posterior to the IOL requiring a second PPV and one eye developed pseudophakic glaucoma. Two patients required a third PPV to keep the visual axis clear. In group B, 0 of 16 (P <.0001) developed secondary opacification of the visual axis. In group C, 4 of 33 (12%; P <.0001) developed pupillary opacification in the first postoperative year. CONCLUSIONS: Intraocular lens implants in infants may be associated with a higher complication rate requiring further surgery during the first postoperative year than is lensectomy/vitrectomy surgery without IOL implant in infants or lensectomy/IOL/vitrectomy surgery in children older than 6 months of age. PMID- 11907474 TI - Factors associated with horizontal reoperation in infantile esotropia. AB - PURPOSE: Risk factors for requiring multiple surgeries in infantile esotropia remain unclear. We identified clinical and demographic factors associated with horizontal reoperation in this disease. METHODS: A retrospective chart review of patients who underwent surgery from 1994-1997 was performed. Subjects were divided into 2 groups: those requiring only one operation and those requiring 2 or more operations to achieve orthotropia +/-10 PD. RESULTS: In 149 patients, the overall horizontal reoperation rate was 34%. There were no statistically significant differences between the 2 groups with respect to mean age at first surgery, mean preoperative deviation, gender, prematurity, Medicaid coverage, parental age, family history of strabismus, or refractive error. The presence of nystagmus, oblique muscle dysfunction, dissociated vertical deviation (DVD), or a variable angle of esotropia was not associated with increased horizontal reoperation rate. There was a greater frequency of horizontal reoperation in patients with amblyopia, although not significant. Premature infants and infants with neurologic dysfunction had a lower incidence of horizontal reoperation, but also not significant. Deviations of less than 30 PD were associated with fewer horizontal reoperations (16% vs 31%, P =.047). Significantly more patients underwent horizontal reoperation when initial surgery was performed at less than or equal to 15 months of age (67% vs. 47%, P =.022). CONCLUSIONS: Several factors thought to predispose to poor sensorimotor outcome (dissociated vertical deviation, oblique muscle dysfunction, and nystagmus) were not associated with an increased incidence of horizontal reoperation. Horizontal reoperation was less frequent in patients with angles less than 30 PD. Although some studies suggest that early surgical intervention in patients with infantile esotropia affords better sensory outcome, it may be associated with a higher horizontal reoperation rate. PMID- 11907475 TI - Bilateral lateral rectus resection for residual esotropia. AB - PURPOSE: Residual or recurrent esotropia is a common problem following bilateral medial rectus recessions for esotropia, and various surgical techniques have been advocated. We have favored bilateral lateral rectus resections. METHODS: We reviewed our results in 25 patients, aged 7 to 89 months (mean 27 months), with a follow-up of 7 to 95 months (mean 39 months) following the second surgery. Survival analysis was used, with success (survival) defined as alignment within 10 PD at last follow-up. RESULTS: Median survival was 84 months with an estimated mean survival of 55 months. There are 15 of 25 patients (60%) currently successful. There were 8 undercorrections and 2 overcorrections. Three patients, included among those not successful, required a third procedure. Survival was similar to previous reports of primary esotropia and consecutive exotropia. CONCLUSIONS: Our results suggest that bilateral lateral rectus resection is a reasonable surgical option in the treatment of residual esotropia following recessions of both medial recti. PMID- 11907476 TI - Retinoblastoma in older children. AB - PURPOSE: Retinoblastoma (RB) is known to present with atypical signs and symptoms in older patients. Our article reviews the clinical and histopathological features of RB encountered in children older than 5 years of age. METHODS: A total of 337 consecutive patients with RB were reviewed. Eighteen (5.3%; 10 males, 8 females) who presented after the age of 5 were selected for further analysis of clinical data and histopathological material in this retrospective, non-comparative case series. RESULTS: The age range was from 5 to 12 years (mean age 6.4). Unilateral disease was seen in 14 children, bilateral disease in 4 children. Eleven children were the product of consanguineous marriages. Seven patients had leukocoria; the others presented with signs of uveitis, cellulitis, and/or trauma. Three patients had flat, plaque-like lesions on histopathologic evaluation. CONCLUSIONS: Clinical and histopathologic features were atypical in 47% and 21% respectively in the retinoblastoma patients who presented after the age of 5 years. PMID- 11907477 TI - 20 unusual presentations of accommodative esotropia. AB - PURPOSE: This paper will discuss several unusual presentations of accommodative esotropia. METHODS: A total of 20 patients with unusual case histories who presented with accommodative esotropia were studied retrospectively. These patients were separated into 3 categories. The first contained children who had their onset of accommodative esotropia after a traumatic event such as head trauma or ocular trauma. The second involved infants between 3 and 5 months of age who presented with accommodative esotropia. The third showed the onset of accommodative esotropia associated with diabetic ketoacidosis. (There were no other metabolic disorders associated with accommodative esotropia.) RESULTS: All patients resolved their esotropia with glasses initially but 2 did require surgery several years after the onset of the esotropia. CONCLUSIONS: Children under 5 months of age with intermittent esotropia and/or significant amounts of hyperopia should have the diagnosis of accommodative esotropia considered as the etiology of their esotropia. Children with the onset of esotropia associated with trauma who have significant amounts of hyperopia should also have accommodative esotropia considered as an etiology of their crossing. PMID- 11907478 TI - Eye dominance in the visual cortex using functional MRI at 1.5 T: an alternative method. AB - PURPOSE: To develop a functional MRI method for producing eye dominance histograms in humans at 1.5 Tesla (T). METHODS: In the first set of experiments, 8 normal persons were tested. The eye dominance of each voxel within the person's visually activated primary visual cortex was determined with Student t statistics during a left eye versus right eye contrast. Eye dominance distribution was plotted, and the mean t statistic was used to describe the histogram asymmetry. In the second set of experiments, the effect of monocular optical blur and decreased luminance via filter was studied, and eye dominance distributions were similarly determined. RESULTS: The eye dominance histogram in each of the 8 normals was approximately symmetric; the average mean t value was +0.13. All 4 subjects with the right eye blurred had histograms approximately symmetric or slightly shifted toward the left eye (average mean t = +0.56), and all 4 subjects with the right eye filtered had histograms dramatically shifted toward the left eye (average mean t = +2.22). The average mean t for the group with the right eye filtered was significantly different from that of the other 2 groups (P <.0001). CONCLUSIONS: With noninvasive methods in normal persons, functional magnetic resonance imaging techniques at 1.5 T were able to characterize the distribution of eye dominance of voxels in primary visual cortex, based upon their t statistic in the left eye versus right eye contrast. The method is sensitive to filtering but relatively insensitive to visual blur. This approach may have a future use in the study of amblyopia in humans. PMID- 11907479 TI - Optic disc Drusen associated with trisomy 15q. PMID- 11907480 TI - Inferior oblique muscle palsy following maxillectomy for squamous cell carcinoma. PMID- 11907481 TI - Orbital neural-glial hamartoma associated with a congenital tonic pupil. AB - Tonic pupils in children are rare, and only a handful of reports exist in the literature. We present the case of an orbital neural-glial hamartoma in an infant with a congenital tonic pupil and no proptosis. We have previously described this association in another child. This new case, the 6-year follow-up from the previous case, and a discussion of tonic pupils are presented. PMID- 11907482 TI - Photodisruption of dense preretinal hemorrhage with Nd:YAG in a child with Terson's syndrome. PMID- 11907483 TI - Surgical outcome after prism adaptation for esotropia with a distance-near disparity. PMID- 11907485 TI - Changes in plasma protein binding have little clinical relevance. PMID- 11907486 TI - Pharmacokinetics and pharmacodynamics of saquinavir in pediatric patients with human immunodeficiency virus infection. AB - OBJECTIVE: Our objective was to investigate the clinical pharmacologic characteristics of saquinavir given as a soft gelatin capsule, either alone or in combination with nelfinavir, to children and adolescents with human immunodeficiency virus infection. METHODS: The pharmacokinetics of 50 mg/kg saquinavir 3 times a day (tid) alone versus 33 mg/kg saquinavir tid plus 30 mg/kg nelfinavir tid was assessed after single-dose administration and after short- and long-term administration. The single-dose pharmacokinetics of fixed (1200 mg) versus unrestricted weight-adjusted dosing (50 mg/kg) was also investigated. RESULTS: Saquinavir as the sole protease inhibitor resulted in lower saquinavir exposure in children (steady-state geometric mean area under the concentration time curve from time zero to 24 hours [AUC (0-24 h)], 5790 ng x h/ml; steady state concentration 8 hours after drug administration [C(8h,SS)], 65 ng/ml) and adolescents [steady-state geometric mean AUC(0-24 h), 5914 ng x h/ml] than that reported in adults treated with 1200 mg tid [steady-state geometric mean AUC(0-24 h), 21,700 ng x h/ml; C(8h,SS), 223 ng/ml]. This finding appeared to be attributable to markedly higher apparent oral clearance, potentially as a result of increased systemic clearance and reduced oral bioavailability. Nelfinavir combined with saquinavir reduced apparent oral clearance, increasing saquinavir exposure in children [steady-state geometric mean AUC(0-24 h), 11,070 ng x h/ml; C(8h,SS), 380 ng/ml] to levels that approach those observed in adults. A significant correlation between average trough concentration and sustained viral load suppression was observed in children. The apparent threshold for maintaining viral load suppression was a mean trough saquinavir concentration above 200 ng/ml. CONCLUSIONS: The pharmacokinetics of saquinavir in children is different from that of adults, and administration of saquinavir alone will not give consistently efficacious plasma levels. The best way of improving saquinavir exposure in children is through combination therapy with other protease inhibitors that inhibit saquinavir metabolism. PMID- 11907487 TI - Shed human enterocytes as a tool for the study of expression and function of intestinal drug-metabolizing enzymes and transporters. AB - OBJECTIVES: Intestinal metabolism and transport are now recognized as protective barriers against orally ingested xenobiotics, including drugs. However, in vitro studies of the expression and function of intestinal proteins are hampered by the limited availability of human intestinal tissues. Because enterocytes are constantly shed in large numbers into the gut lumen, this study investigated whether these cells could be collected with a multilumen perfusion catheter and whether they are functionally active. METHODS: In healthy volunteers, a 20-cm isolated jejunal segment was generated with the perfusion catheter by inflating 2 balloons with air. Shed cells were characterized by fluorescence-activated cell sorting analysis for leukocyte-specific CD45 and enterocyte-specific villin, as well as for apoptosis. Homogenates of the cells were used for reverse transcriptase polymerase chain reaction and Western blotting. Cytochrome P450 enzyme activity was determined with the calcium channel blocker verapamil as a substrate. RESULTS: On average, 4.83 mg protein and 56.23 million cells were collected from a 20-cm segment during 2 hours. A total of 84.2% of the cells were positive for enterocyte-specific villin, and only 1.6% of the collected cells were positive for CD45. The majority of cells (65.3%) were not in early or late apoptosis or necrosis. In all volunteers, drug-metabolizing enzymes (such as members of the cytochrome P450 family) could be detected as both messenger ribonucleic acid and proteins. Consistent with expression data, formation of verapamil metabolites catalyzed by CYP3A4 and CYP2C was shown. CONCLUSIONS: The majority of shed human enterocytes collected with a multilumen perfusion catheter were still functionally active and not apoptotic. Harvesting of spontaneously shed enterocytes provides a new tool for studies on expression and function of intestinal proteins. PMID- 11907488 TI - Low daily 10-mg and 20-mg doses of fluvoxamine inhibit the metabolism of both caffeine (cytochrome P4501A2) and omeprazole (cytochrome P4502C19). AB - OBJECTIVES: Fluvoxamine is metabolized by the polymorphic cytochrome P450 (CYP) 2D6 and the smoking-inducible CYP1A2. Therapeutic doses of fluvoxamine inhibit both CYP1A2 and CYP2C19. In this study we used extensive metabolizers (EMs) and poor metabolizers (PMs) of debrisoquin (INN, debrisoquine) (CYP2D6) and two probes, caffeine (CYP1A2) and omeprazole (CYP2C19), to investigate whether nontherapeutic doses of fluvoxamine inhibit CYP1A2 but possibly not CYP2C19. METHODS: Single oral doses of 100 mg caffeine and 20 mg omeprazole were given separately to 5 EMs and 5 PMs of debrisoquin to assess the activity of CYP1A2 and CYP2C19, respectively. Initially, a single oral dose of fluvoxamine (25 mg to PMs and 50 mg to EMs) was given, followed by 1 week of daily administration of 25 mg x 2 to EMs and 25 mg x 1 to PMs. Caffeine (day 6) and omeprazole (day 7) were again administered at the steady state of fluvoxamine. Later the study protocol was repeated with a lower dose of fluvoxamine, 10 mg x 2 to EMs and 10 mg x 1 to PMs for 1 week. Concentrations of fluvoxamine, caffeine, omeprazole, and their metabolites were analyzed by HPLC methods in plasma and urine. RESULTS: The kinetics of fluvoxamine were not significantly different in EMs and PMs after a single oral dose of the drug. At the higher but not the lower steady-state dose of fluvoxamine, a significantly lower clearance in PMs compared with EMs was observed (geometric mean, 0.86 versus 1.4 L/h per kilogram; P <.05). At steady state, the 25 mg x 1 or x 2 fluvoxamine dose caused a pronounced inhibition of about 75% to 80% for both CYP1A2 and CYP2C19, whereas the inhibition after the lower 10 mg x 1 or x 2 dose was about 40% to 50%. The area under the plasma concentration-versus-time curve from 0 to 24 hours [AUC(0-24)] of caffeine increased 5-fold (P <.001) after the higher dose of fluvoxamine and 2-fold (P <.05) after the lower dose. The area under the plasma concentration-time curve from time zero to 8 hours [AUC(0-8)] ratio of 5-hydroxyomeprazole/omeprazole decreased 3.4-fold (P <.001) and 2.4-fold (P <.001), respectively. One EM subject had a very low oral clearance of fluvoxamine after both single and multiple dosing of the drug. This subject might have a deficient transporter protein in the gut, leading to an increased absorption of fluvoxamine. CONCLUSION: No convincing evidence was found that CYP2D6 is an important enzyme for the disposition of fluvoxamine. Other factors seem to be more important. A nontherapeutic oral daily dose of fluvoxamine is sufficient to provide a marked inhibition of both caffeine (CYP1A2) and omeprazole (CYP2C19) metabolism. It was not possible to separate the inhibitory effects of fluvoxamine on these enzymes, even after such a low daily dose such as 10 mg x 1 or x 2 of fluvoxamine. PMID- 11907489 TI - Time course of 8-methoxypsoralen concentrations in skin and plasma after topical (bath and cream) and oral administration of 8-methoxypsoralen. AB - BACKGROUND: The combination of 8-methoxypsoralen with ultraviolet A exposure (PUVA therapy) is a standard treatment for a variety of dermatoses. The following three variants have been described: oral, bath, or cream PUVA. To achieve optimal therapeutic effects, ultraviolet A irradiation should be performed at the time of maximum photosensitivity, that is, at the time of maximum 8-methoxypsoralen tissue concentrations. METHODS: To further specify this point of time, we assessed the concentration-time courses of 8-methoxypsoralen in the skin after oral, bath, and cream administration of 8-methoxypsoralen in a 3-way crossover microdialysis study of 8 healthy subjects. RESULTS: Tissue concentrations after oral administration of 0.6 or 1 mg/kg 8-methoxypsoralen were low (peak plasma concentration range, 1.7-6.6 ng/ml) compared with topical administration for which maximum concentrations of 200 to 520 ng/ml and 720 to 970 ng/ml were achieved with 0.1% 8-methoxypsoralen cream and 3 mg/L 8-methoxypsoralen bath, respectively. Plasma concentrations after oral 8-methoxypsoralen, however, were up to 1000-fold higher than those found after topical application. With both topical applications, the tissue peak concentration uniformly occurred in the first 20 minutes after the end of the application time. In contrast, the time to reach the tissue peak concentration after oral administration ranged from 1 to 4 hours. CONCLUSIONS: The time course of tissue concentrations corresponds closely with the time course of minimal phototoxic doses found in previous studies. Because tissue concentrations after topical administration of 8-methoxypsoralen (bath and cream) were high compared with plasma concentrations and because they were less variable and occurred at better predictable time points than those after oral administration, we suggest that topical PUVA is superior to systemic PUVA, at least from a pharmacokinetic point of view. PMID- 11907490 TI - Effect of growth hormone on hepatic cytochrome P450 activity in healthy elderly men. AB - OBJECTIVE: Our objective was to study the effect of recombinant human growth hormone (rhGH) on hepatic cytochrome P450 (CYP) activity in 30 healthy elderly men. METHODS: The study was carried out as a randomized, double-blind, placebo controlled parallel-group study. rhGH or placebo was administered for a period of 12 weeks. CYP activity was measured before, after 12 weeks of rhGH and placebo administration, and at 4 weeks after termination of rhGH and placebo administration with use of the biomarker reactions of CYP1A2 (caffeine), CYP2C19 (mephenytoin), CYP2D6 (sparteine), CYP3A4 (endogenous cortisol metabolism), and antipyrine clearance as common markers of CYP activity. RESULTS: The metabolic ratio of caffeine increased significantly in the group that received growth hormone compared with placebo (median difference, 4.55; 95% confidence interval (CI), 1.64 to 8.60; versus -0.90; 95% CI, -5.70 to 1.36), indicating an induction of CYP1A2. Moreover, the S/R ratio of mephenytoin showed a small but significant increase (median difference, 0.02; 95% CI, 0 to 0.31; versus 0; 95% CI, -0.01 to 0.06), indicating an inhibition of CYP2C19. There were no significant changes of the metabolic ratios of cortisol and sparteine or the antipyrine clearance compared with placebo. CONCLUSIONS: These results indicate that growth hormone induces CYP1A2 and, to a lesser extent, inhibits CYP2C19 in elderly men, but it exerts no effects on CYP2D6 and CYP3A4. Although the induction of CYP1A2 may be of some clinical relevance, the small inhibition of CYP2C19 is probably unimportant. PMID- 11907491 TI - Cardiovascular effect of almotriptan in treated hypertensive patients. AB - OBJECTIVE: Our objective was to investigate the cardiovascular effects of almotriptan, a 5-hydroxytryptamine 1B/1D agonist, in treated patients with hypertension. METHODS: Twenty patients with hypertension controlled by medication received the following treatments in a randomized, double-blind, crossover design: one placebo tablet, one 12.5-mg almotriptan tablet, and one 25-mg almotriptan tablet. Serial blood samples for analysis of almotriptan were obtained through 24 hours after administration. Serial measurements of supine blood pressure, 12-lead electrocardiograms, and Holter electrocardiographic recordings were also obtained. Plasma almotriptan concentrations were measured with use of a liquid chromatography-tandem mass spectrometry assay. Differences between treatments in pharmacokinetic parameters were assessed with an ANOVA model appropriate for a crossover design. Blood pressure measures and QTc intervals were analyzed for treatment effects with use of a similar model. Analyses were performed on weighted mean and maximal changes from baseline for intervals from 0 to 4 and 0 to 12 hours after administration. RESULTS: Significant linear effects of dose were observed for the maximal change in diastolic blood pressure and for the maximal and mean changes in systolic blood pressure. These effects were consistent for both the 4- and 12-hour periods after dosing. Mean changes from baseline during the interval from 0 to 4 hours were 1.59 +/- 3.88, 1.85 +/- 5.94, and 4.84 +/- 5.99 mm Hg for systolic pressure and 1.38 +/- 6.95, 6.25 +/- 9.54, and 11.0 +/- 10.6 mm Hg for diastolic pressure for placebo, 12.5 mg almotriptan, and 25 mg almotriptan, respectively. No instances of hypertensive crisis were observed. No QTc interval prolongation was observed. CONCLUSIONS: Almotriptan has effects on blood pressure in subjects with controlled hypertension that are consistent with those of other members of the pharmalogic class. PMID- 11907492 TI - Clopidogrel, but not abciximab, reduces platelet leukocyte conjugates and P selectin expression in a human ex vivo in vitro model. AB - Formation of platelet-leukocyte aggregates (PLA) through the CD62 ligand is an important mechanism by which leukocytes contribute to thrombosis and inflammation. We investigated the formation of PLA in human subjects after stimulation with thrombin receptor activating peptide and adenosine diphosphate (ADP) after treatment with clopidogrel and after in vitro application of the platelet glycoprotein IIb/IIIa complex antagonist abciximab. Expression of CD62 was significantly reduced 30% to 50% with clopidogrel, depending on the type and concentration of the inducer, but addition of abciximab led to a significant approximately 30% increase in CD62 expression when platelets were stimulated by ADP. Formation of PLA decreased significantly with clopidogrel to 55% to 75% of the baseline value, whereas addition of abciximab caused a significant increase in PLA in ADP-stimulated samples before but not after administration of clopidogrel. The increase in formation of PLA after in vitro addition of abciximab was not paralleled by a decrease in platelet microaggregates and is therefore presumed not caused by enhanced availability of platelets. To our knowledge, this is the first report showing that clopidogrel reduces formation of PLA. The findings also suggest intersection between an "outside-in" signal generated by abciximab and stimulation of platelet P2T(12) purinergic receptors that augments degranulation and increases formation of PLA but is inhibited by clopidogrel. PMID- 11907494 TI - CYP3A4 variant alleles in white individuals with low CYP3A4 enzyme activity. AB - OBJECTIVE: Our objective was to evaluate the presence of CYP3A4 gene variants in white individuals with low CYP3A4 enzyme activity. METHODS: Persons with extremely low enzyme activity, either in vitro or in vivo, were selected in a panel of 97 healthy subjects. Genetic analyses for CYP3A4 variant alleles present in white subjects, including CYP3A4*1B, CYP3A4*2, CYP3A4*4, CYP3A4*5, CYP3A4*6, CYP3A4*8, CYP3A4*11, CYP3A4*12, and CYP3A4*13, were performed on genomic deoxyribonucleic acid from these subjects by amplification-restriction and sequencing. RESULTS: With the exception of CYP3A4*1B, none of the variant alleles analyzed were present in 30 genes from persons with extremely low enzyme activity. CYP3A4*1B was present in the population studied with an allele frequency of 5.5%. Nevertheless, the presence of CYP3A4*1B does not correlate with low enzyme activity, either in vivo or in vitro, in either heterozygosity or homozygosity. CYP3A4*2 was not identified in 290 genes from Spanish persons or in 70 genes from Finnish persons. CONCLUSIONS: Although the genetic component of the interindividual variability of CYP3A4 enzyme activity seems to be high, our findings do not support a key role for the variant alleles analyzed on the majority of white persons with low CYP3A4 activity. This suggests the occurrence of as yet unknown mutations that affect CYP3A4 or other functionally related genes. PMID- 11907493 TI - Pharmacodynamics of abciximab during angioplasty: comparison to healthy subjects. AB - OBJECTIVES: Our objectives were to compare and contrast abciximab concentration effect relationships in healthy volunteer participants with those in patients with coronary atherosclerosis undergoing elective coronary angioplasty. We also aimed to establish abciximab plasma concentrations associated with 80% inhibition of platelet aggregation. METHODS: Abciximab clearance and concentration-effect relationships were determined from two separate clinical studies, one in 30 healthy subjects aged 21 to 66 years and the other in 32 patients aged 44 to 74 years before they underwent elective coronary angioplasty. After abciximab administration, abciximab plasma concentrations, platelet glycoprotein IIb/IIIa (GP IIb/IIIa) receptor occupancy, and degree of inhibition of platelet aggregation in the presence of 5-micromol/L and 20-micromol/L adenosine diphosphate was determined. With an E(max) (receptor occupancy) or inhibitory E(max) (inhibition of platelet aggregation) model, abciximab concentrations required for 80% receptor occupancy and 80% inhibition of platelet aggregation were determined. RESULTS: Abciximab steady-state clearance in healthy participants was 183 +/- 72 ml/min (mean +/- SD), and single-dose clearance in patients undergoing angioplasty was 405 +/- 240 ml/min (mean +/- SD). Abciximab concentration required for 80% GP IIb/IIIa receptor occupancy was 35.2 +/- 2.4 versus 72.8 +/- 6.4 ng/ml in healthy participants versus patients (P <.01). Concentrations required for 80% inhibition of platelet aggregation stimulated by 5-micromol/L adenosine diphosphate were 25.6 +/- 1.6 versus 68.9 +/- 9.2 ng/ml (P <.01). Similarly, the concentrations required for 80% inhibition of platelet aggregation stimulated by 20-micromol/L adenosine diphosphate were 56.0 +/- 3.2 versus 141 +/- 16.8 ng/ml (P <.01). CONCLUSION: Approximately 2-fold greater abciximab exposure is required to achieve the same degree of GP IIb/IIIa occupancy and inhibition of platelet aggregation in patients undergoing angioplasty as compared with healthy participants. The difference between groups may be related either to different states of basal platelet activation or to the effect of heparin that patients received as part of the angioplasty procedure. A therapeutic concentration range for patients is 100 to 175 ng/ml, because this is the concentration consistent with >80% inhibition of platelet aggregation when 20 micromol/L adenosine diphosphate is used as the aggregating stimulus. PMID- 11907496 TI - Cutaneous smooth muscle neoplasms: clinical features, histologic findings, and treatment options. AB - Cutaneous smooth muscle is present in 3 separate locations: arrector pili muscles, blood vessel walls, and genital/areolar skin. Benign or malignant smooth muscle neoplasms may arise from each of these locations. This review discusses the pathogenesis, clinical manifestations, histologic findings, prognosis, treatment options, and controversial areas of cutaneous smooth muscle neoplasms. ( J Am Acad Dermatol 2002;46:477-90.) LEARNING OBJECTIVE: At the completion of this learning activity, participants should be able to discuss the pathogenesis, clinical manifestations, histologic findings, prognosis, and treatment options of cutaneous smooth muscle neoplasms. PMID- 11907497 TI - Safety and efficacy of pimecrolimus (ASM 981) cream 1% in the treatment of mild and moderate atopic dermatitis in children and adolescents. AB - BACKGROUND: The ascomycin derivative pimecrolimus (ASM 981) is a cell-selective cytokine inhibitor, specifically developed for the treatment of inflammatory skin diseases. OBJECTIVE: When applied topically, pimecrolimus cream 1% has shown promise as a treatment for inflammatory skin conditions, including atopic dermatitis (AD) in children and adults, allergic contact dermatitis, and chronic contact irritant hand dermatitis in adults. METHODS: In two independent 6-week, randomized, multicenter studies of identical design, the efficacy and safety of pimecrolimus cream 1% in children with predominantly moderate AD were compared with vehicle. Pooled data from a total of 403 patients were used in the analysis. The primary efficacy parameter was the Investigator's Global Assessment (IGA) score. Secondary parameters included Eczema Area and Severity Index (EASI) and severity of pruritus scores. Subjects were also asked to assess their disease control as uncontrolled, limited, good, or complete. RESULTS: Significant therapeutic benefits relative to vehicle were observed in the pimecrolimus treated group at the first efficacy assessment, 8 days after initial application of the study medication (eg, relief of pruritus). At each subsequent postbaseline visit, pimecrolimus-treated patients showed significant improvement relative to controls in all efficacy measures. The medication was well tolerated. CONCLUSION: Pimecrolimus cream 1% appears to be a safe and effective alternative to currently used therapies for AD. PMID- 11907498 TI - Use of isotretinoin (Accutane) in the United States: rapid increase from 1992 through 2000. AB - BACKGROUND: Isotretinoin, a drug approved to treat severe recalcitrant nodular acne, has been marketed in the United States since 1982. The drug is an effective treatment for acne that is refractory to other therapies, but it is a teratogen and can cause serious side effects. OBJECTIVE: Our purpose was to describe trends in the use of isotretinoin in the United States from marketing through year 2000 and summarize characteristics of patients and prescribers. METHODS: Data from 2 pharmaceutical marketing research databases, the National Prescription Audit Plus and the National Disease and Therapeutic Index, and from 2 health plan networks were obtained and analyzed. RESULTS: Retail pharmacies dispensed 19.8 million outpatient prescriptions for isotretinoin from marketing in 1982 through 2000. From 1983 through 1993, the median annual number of prescriptions was just over 800,000; between 1992 and 2000, the number of prescriptions increased 2.5-fold (250%) to nearly 2 million in year 2000. The increases registered in the health plans were somewhat larger: about 275% increases from 1995 through 1999. There is no ICD-9 code for nodulocystic acne; consequently, the type of acne treated with isotretinoin is not determinable from these data. However, between 1993 and 2000, the proportion of isotretinoin treatment for severe acne declined from 63% to 46%, whereas the proportion of treatment for mild and moderate acne increased from 31% to 49%. Data also indicated that the sex distribution of patients was nearly even, and that 63% of male patients prescribed isotretinoin were 15 to 19 years old, whereas 51% of female patients were 15 to 24 years old. CONCLUSION: In the last 8 years, there has been a 2.5-fold (250%) increase in the number of dispensed prescriptions for isotretinoin in the United States. Data also reveal an increasing proportion of isotretinoin use for mild and moderate acne. PMID- 11907499 TI - Junctional epidermolysis bullosa in the Middle East: clinical and genetic studies in a series of consanguineous families. AB - BACKGROUND: Junctional epidermolysis bullosa (JEB) is a group of inherited blistering diseases characterized by epidermal-dermal separation resulting from mutations that affect the function of critical components of the basement membrane zone. This group of autosomal recessive diseases is especially prevalent in regions where consanguinity is common, such as the Middle East. However, the clinical and genetic epidemiology of JEB in this region remains largely unexplored. OBJECTIVE: The aim of the present study was to assess a series of consanguineous JEB families originating from the Middle East. METHODS: We identified 7 families referred to us between 1998 and 1999 and originating from the United Arab Emirates, Saudi Arabia, Sudan, Yemen, and Israel. Histologic, immunofluorescence, and electron microscopy studies were performed to direct the subsequent molecular analysis. DNA obtained from all family members was amplified by means of polymerase chain reaction and analyzed by conformation-sensitive gel electrophoresis with subsequent direct sequencing. RESULTS: In 6 families presenting with the clinical and histologic features distinctive for JEB, mutations in genes encoding 1 of the 3 subunit polypeptides of laminin-5 were identified. Two families each had mutations in LAMB3, 2 in LAMA3, and 2 in LAMC2. Out of 7 distinct mutations, 5 were novel and 2 were recurrent. No relationship was found between the presence of nonsense/frameshift mutations in laminin-5 genes and perinatal mortality, contradicting a major genotype-phenotype correlation previously reported in the European and US literature. Similarly, none of the recurrent LAMB3 hot spot mutations previously described in other populations was found in our series. Finally, in a family with the clinical diagnosis of generalized atrophic benign epidermolysis bullosa, a homozygous non sense mutation in Col17A1 gene (encoding the BPAG2 antigen) was identified. CONCLUSION: The present report suggests (1) the existence of a unique spectrum of mutations in the Middle East populations and (2) the need for the implementation of a diagnostic strategy tailored to the genetic features of JEB in this region. PMID- 11907500 TI - Changes in hair weight and hair count in men with androgenetic alopecia after treatment with finasteride, 1 mg, daily. AB - BACKGROUND: Finasteride, a type II 5alpha-reductase inhibitor, reduces scalp and serum dihydrotestosterone and has been shown to be effective in men with androgenetic alopecia (AGA). OBJECTIVE: The purpose of this study was to determine the effect of finasteride on scalp hair weight in men with AGA. METHODS: Sixty-six men with AGA received finasteride, 1 mg/d, or placebo in a 48 week study, and 49 men continued in a 48-week extension. Efficacy was assessed by scalp hair weights and hair counts. RESULTS: As expected, hair counts improved with finasteride (net mean percent change +/- SE [95% CI] compared with placebo = 9.2% +/- 2.8% [3.8, 14.6] and 15.4% +/- 3.2% [9.1, 21.7] at 48 and 96 weeks, respectively; P <.01 for both time points), and net improvements in hair weight were greater (25.6% +/- 3.6% [18.5, 32.7] and 35.8% +/- 4.6% [26.7, 44.8] at 48 and 96 weeks, respectively; P <.001 for both time points). Finasteride was generally well tolerated. CONCLUSION: In this study, finasteride, 1 mg, increased hair weight in men with AGA. Hair weight increased to a larger extent than hair count, implying that factors other than the number of hairs, such as increased growth rate (length) and thickness of hairs, contribute to the beneficial effects of finasteride in treated men. PMID- 11907501 TI - Human chronic wounds treated with bioengineered skin: histologic evidence of host graft interactions. AB - Bioengineered skin is being used to successfully treat a variety of wounds. Randomized controlled clinical trials have shown that a living bilayered skin construct (BSC), consisting of human neonatal keratinocytes and fibroblasts in a collagen matrix, was able to accelerate complete closure of both venous and diabetic ulcers. BSC was particularly effective in difficult-to-heal wounds of long duration. In patients treated with BSC, no obvious signs of gross clinical rejection were observed. Testing of these treated patients showed no BSC-specific immune response and no immune response to bovine collagen or alloantigens expressed on keratinocytes and fibroblasts. However, very little is known about the histologic changes that occur after BSC has been placed on human wounds. We report our preliminary histologic observations in this uncontrolled study of a cohort of 11 patients with 14 wounds treated with BSC in whom biopsy specimens of the grafted sites were obtained at least 2 weeks after application of the construct. The etiology of these ulcers varied from arterial or venous disease to an extensively and poorly healing burn wound. Histologically, thickening of the grafted bioengineered skin was seen in all samples where residual BSC could be identified. Mucin deposition was noted in the dermal layer of the wounds and BSC in 13 of the 14 specimens examined. Unexpectedly, and in spite of good clinical outcome, 4 of the 14 specimens exhibited a foreign body-like granulomatous response. There was no history of prior exposure to BSC in the 4 patients who had a granulomatous response. These early histologic observations suggest that stimulatory interactions develop between BSC and the wound. The consistently found deposition of mucin may point to a fetal pattern of wound repair associated with the neonatal cells in BSC. PMID- 11907502 TI - Localized cutaneous small to medium-sized pleomorphic T-cell lymphoma: a report of 3 cases stable for years. AB - Small to medium-sized pleomorphic cutaneous T-cell lymphomas represent a provisional entity in the new European Organization for Research and Treatment of Cancer classification. We describe 3 patients with a localized and outstanding stable variant of this tumor. A median follow-up period of 50 months did not reveal any spread into regional lymph nodes or to distant sites in any patient. PMID- 11907503 TI - High-dose botulinum toxin type A therapy for axillary hyperhidrosis markedly prolongs the relapse-free interval. AB - BACKGROUND: Axillary hyperhidrosis is a common condition that can be personally distressing and can interfere with professional and social life. Intracutaneous injections of botulinum toxin type A (BTXA) have recently been shown to induce an effective but temporary anhidrosis, usually for 4 to 6 months. High-dose BTXA was shown to have a lower relapse rate, but it remained unclear whether it could induce a prolongation of the antihidrotic effect. OBJECTIVE: Our aim was to evaluate the long-term effectiveness of "high-dose" botulinum toxin therapy in axillary hyperhidrosis, the response to repeated treatment, and the possible side effects. METHODS: In an open study in a university medical center, 47 patients with axillary hyperhidrosis unresponsive to previous therapies were treated with intracutaneous injections of botulinum A toxin. A total dose of 200 U of BTXA was used per axilla. Patients were followed up for periods up to 29 months. The main outcome measures were patients' satisfaction with the antihidrotic effect, response to repeated treatment, and safety of treatment. RESULTS: Within 6 days of the injection of BTXA, all patients reported cessation of excessive sweating. The follow-up was 17.0 +/- 8.3 months (range, 3-29 months). The relapse rate within 12 months of treatment was 4 of 34 patients (11.8%). The longest relapse free interval observed was 29 months. Twenty-two patients showed a relapse-free interval of 19 months or more. The number of patients with at least 12 months of remission was significantly higher with 200 U of botulinum toxin than with lower doses reported in current literature (P <.05). Relapsed patients (a total of 6/47) showed an unchanged excellent response to a second or third treatment. The only side effect was temporary pain and burning during the injections. No muscular weakness, insensitivity, or systemic reactions were observed. CONCLUSION: High-dose BTXA treatment is capable of prolonging the antihidrotic effect of intracutaneous BTXA in most patients for more than 19 months. It reduces the percentage of relapses after 12 months of follow-up to less than 12%. So far, there is no clinical evidence of the induction of neutralizing antibodies. High-dose treatment seems to be as safe as low-dose treatment. PMID- 11907504 TI - Sulfasalazine for alopecia areata. AB - Sulfasalazine is used as a therapy for various autoimmune conditions, including psoriasis; its effectiveness is presumed to be the result of its immunomodulatory effects. We have treated patients with severe alopecia areata with sulfasalazine as part of our dermatology practice and have noticed cosmetically acceptable regrowth in 23% of patients in whom a response could be determined. In view of its good safety profile, sulfasalazine may be considered for systemic treatment of severe alopecia areata. PMID- 11907505 TI - Squamous cell carcinoma in situ of the penis successfully treated with imiquimod 5% cream. AB - BACKGROUND: Multiple treatments for squamous cell carcinoma in situ (SCCIS) of the penis have been used with variable success and morbidity. Surgery and destructive treatment modalities have significant risk of scarring, deformity, and impaired function. OBJECTIVE: The purpose of this study was to determine whether topical imiquimod 5% cream is a potentially effective treatment for SCCIS of the penis and to qualify treatment associated morbidity. METHODS: The case of a patient with extensive penile SCCIS is reported. The patient was treated with topical imiquimod 5%, administered daily until blistering occurred (2 cycles). Biopsy specimens were obtained to confirm tumor clearance. RESULTS: One month after therapy was completed, no clinical or histologic evidence of residual tumor was found. Adverse effects of imiquimod included localized tenderness and erythema. No evidence of scarring, deformity, loss of function, or tumor recurrence was noted 18 months after treatment. CONCLUSION: Imiquimod 5% cream may represent an alternative treatment option for SCCIS of the penis. PMID- 11907506 TI - The significance of tumor persistence after incomplete excision of basal cell carcinoma. AB - BACKGROUND: Physicians inevitably receive a pathology report after excision of a basal cell carcinoma that indicates that it is incompletely excised. The physician and patient are then left with the dilemma of whether immediate re excision or close clinical follow-up is indicated. OBJECTIVE: Our purpose was to identify characteristics of incompletely excised basal cell carcinomas that are at low risk for recurrence. METHODS: We retrospectively reviewed the charts and pathology slides of all incompletely excised basal cell carcinomas from 1991 to 1994 in a university hospital tumor registry. RESULTS: Incompletely excised basal cell carcinomas of superficial or nodular subtype, less than 1 cm in diameter, located anywhere except the nose or ears, with less than 4% marginal involvement on the initial inadequate excision had no evidence of tumor persistence. CONCLUSION: When physicians receive a pathology report indicating the incomplete excision of a basal cell carcinoma, they face the dilemma of further management. The majority of patients should undergo immediate re-excision or Mohs micrographic surgery because tumor persistence was found in 28% of cases. Occasionally, for a small group of select patients, close clinical follow-up may be indicated if the risk of recurrence is very low. PMID- 11907507 TI - A method for pulsed carbon dioxide laser treatment of epidermal nevi. PMID- 11907508 TI - Phototherapy utilization for psoriasis is declining in the United States. AB - Phototherapy is an established treatment modality for psoriasis. The use of phototherapy for psoriasis appears to be in decline in nonfederal and non university-based settings. We used data from the National Ambulatory Medical Care Survey to estimate the number of visits for phototherapy and psoralen ultraviolet A-range (PUVA) light therapy from 1993 to 1998. There were 873,000 visits for UV light therapy in 1993-1994, 189,000 in 1995-1996, and 53,000 in 1997-1998 (P <.0001). There were 175,000 psoralen visits in 1993-1994, 61,000 in 1995-1996, and 25,000 in 1997-1998 (P =.0053). Similar decreases in phototherapy visits occurred in our university-based practice. The decline in phototherapy represents decreased utilization of a safe and effective treatment for psoriasis. PMID- 11907509 TI - Amyopathic dermatomyositis: retrospective review of 37 cases. AB - Criteria for diagnosis of amyopathic dermatomyositis vary, and the prognosis is not clear. Our purpose was to investigate prognosis regarding progression to myositis and associated malignancy. We reviewed the medical records of patients with dermatomyositis evaluated at our institution from 1976 to 1994. Of 746 patients with dermatomyositis, 37 (5%) with the amyopathic subtype were divided into 3 groups: group 1 (73%), no subjective or objective evidence of myopathy; group 2 (13%), no subjective muscle weakness but abnormalities detected by objective tests; group 3 (13%), subjective muscle weakness but no objective evidence of myopathy. Follow-up was conducted by means of a mailed questionnaire. For 25 patients, follow-up of 1 to 17 years after diagnosis showed muscle weakness in 2 patients in group 1 within 5 years after diagnosis. Five patients (13%) had associated malignancies. Of 7 (19%) patients with disease onset before the age of 18 years, none had progression to myopathy. Although it presents with cutaneous lesions indistinguishable from those of classic dermatomyositis, amyopathic dermatomyositis is a distinct entity. In most patients, amyopathic dermatomyositis does not progress to myopathy. Prognosis appears favorable, but malignancy may develop. PMID- 11907510 TI - George Cheyne Shattuck's dissertation on the skin. AB - George Cheyne Shattuck's 1808 dissertation on structure and function of the skin is one of the earliest American writings on the topic of skin diseases. Although this essay contributed no important new observations, it does provide insight into the attitudes of American physicians in the post-Revolutionary War period toward skin diseases, their classification, and their management. PMID- 11907511 TI - Tropical dermatology. Part I. PMID- 11907512 TI - Standard classification of rosacea: Report of the National Rosacea Society Expert Committee on the Classification and Staging of Rosacea. PMID- 11907513 TI - Generalized erythematous macules and plaques associated with flushing, repeated syncope, and refractory anemia. PMID- 11907514 TI - Surgical Pearl: Curettage before cutaneous surgery to identify an unidentifiable biopsy site. PMID- 11907515 TI - A novel missense mutation of NSDHL in an unusual case of CHILD syndrome showing bilateral, almost symmetric involvement. AB - The CHILD syndrome (MIM 308050), an acronym for congenital hemidysplasia with ichthyosiform nevus and limb defects, is an X-linked dominant trait with lethality for male embryos. Recently, we elucidated the underlying gene defect by demonstrating point mutations in NSDHL (NAD[P]H steroid dehydrogenase-like protein) at Xq28 in 6 patients with classic CHILD syndrome. The most striking clinical feature is an inflammatory nevus that usually shows a unique lateralization with strict midline demarcation. Ipsilateral defects involve all skeletal structures and internal organs such as the brain, the lung, the heart, or the kidney. As an exception to this rule, in some cases the CHILD nevus may occur in a more or less bilateral distribution. In 1997 Fink-Puches et al described a case of CHILD nevus with an almost symmetric arrangement. To test the correctness of the diagnosis, we now examined blood lymphocytes of this patient by single-strand conformation analysis and genomic sequencing. We identified a novel missense mutation in NSDHL that potentially may impair protein function. We conclude that a diagnosis of CHILD syndrome can be based on clinical features such as the highly characteristic morphology of the CHILD nevus. A symmetric distribution of this nevus can exceptionally be seen in patients with CHILD syndrome, and this bilateral involvement should not mislead the clinician to any other diagnosis. Apparently, the effect of random X-inactivation is responsible for different patterns of cutaneous involvement in female carriers of NSDHL mutations. PMID- 11907516 TI - Anticonvulsant-induced pellagra. PMID- 11907517 TI - Methotrexate treatment for refractory subacute cutaneous lupus erythematosus. AB - Methotrexate is beneficial in rheumatoid arthritis and has been used in small studies of patients with systemic lupus erythematosus. We describe a patient with severe subacute cutaneous lupus erythematosus refractory to therapy with antimalarials and corticosteroids. Treatment with methotrexate resulted in complete clearing of the skin lesions without any side effects. PMID- 11907518 TI - Lichen planus confined to a radiation therapy site. AB - We report the case of a 58-year-old man with lichen planus localized to a radiation site. To our knowledge, this is the first reported case of radiation induced lichen planus in the English-language literature. PMID- 11907519 TI - Transfer of vitiligo after allogeneic bone marrow transplantation. AB - Adoptive transfer of donor immunity has been demonstrated in animals after bone marrow transplantation (BMT). In humans, several autoimmune diseases have been similarly transferred. Although BMT may, per se, be associated with a modulation of the recipient's immune system, which could trigger or even cause autoimmune diseases, both animal experiments and experience with humans show the likeliness of adoptive transfer of donor immunity to the recipient. We describe a patient with multiple myeloma in whom generalized vitiligo developed within 3 months after allogeneic BMT from his HLA-matched sister with vitiligo. We believe that a form of adoptive transfer of donor immunity to the recipient might play a role in the development of vitiligo. In spite of this, neither de novo development of vitiligo in a genetically predisposed patient nor autoimmune phenomena associated with graft-versus-host disease can be completely excluded as a contributing factor for development of vitiligo in our patient. To our knowledge, this is the first case report of transfer of vitiligo after BMT from a donor with vitiligo. PMID- 11907520 TI - Granulomatous and suppurative dermatitis at interferon alfa injection sites: report of 2 cases. AB - It has previously been reported that interferon alfa injection sites may develop pyoderma gangrenosum, interface dermatitis, vasculitis, or, more commonly, ulcers characterized by intravascular thrombi and a mixed inflammatory cell infiltrate. We describe 2 patients in whom granulomatous and suppurative dermatitis developed at interferon alfa injection sites. These cases extend the spectrum of interferon alfa injection site reactions. The histologic and clinical similarities of these cases with pyoderma gangrenosum and cutaneous Crohn's disease are explored. PMID- 11907521 TI - Botulinum toxin type A injection in the treatment of lichen simplex: an open pilot study. AB - Recalcitrant pruritus is a hallmark of lichen simplex, a localized variant of atopic dermatitis. Acetylcholine has been demonstrated to mediate pruritus in atopic dermatitis. This open pilot study was done to determine the therapeutic effect of blocking acetylcholine release with botulinum toxin A in highly pruritic lichen simplex. Botulinum toxin A (Dysport) was injected intradermally into 5 circumscribed lichenoid lesions in 3 patients suffering from recalcitrant pruritus. No corticosteroids or any other specific topical therapy was administered. Pruritus subsided within 3 to 7 days in all 3 patients. Within 2 to 4 weeks all lesions cleared completely. No recurrences were noted over a 4-month follow-up. In one patient lichen simplex developed on the contralateral shin, which responded equally to a subsequent injection. We concluded that lichen simplex-associated pruritus can be overcome by intradermal botulinum toxin A injection. Acetylcholine appears to be a dominant pruritic mediator in this condition. PMID- 11907522 TI - Persistent depigmented regrowth after alopecia areata. AB - We report a case of persistent depigmented hair regrowth from the site of a patch of alopecia areata. It is well known that hair may regrow unpigmented from a site of alopecia areata; however, it was previously thought that this was temporary, lasting no longer than the first hair cycle. PMID- 11907523 TI - The association of Buschke-Ollendorf syndrome and nail-patella syndrome. AB - Buschke-Ollendorf syndrome and nail-patella syndrome are both rare connective tissue disorders inherited in an autosomal dominant pattern and characterized by cutaneous and bone lesions. We describe a 3-year-old boy and his family who showed clinical features of both Buschke-Ollendorf syndrome and nail-patella syndrome. To our knowledge, this association has not been reported previously, suggesting that these two connective tissue disorders may share the same gene location with different mutations or involve different mutated genes that share downstream segments of their signaling pathways. Furthermore, this young patient is also affected by a chronic idiopathic neutropenia usually not observed in Buschke-Ollendorf syndrome or nail-patella syndrome. PMID- 11907524 TI - Would a new name hasten the acceptance of amyopathic dermatomyositis (dermatomyositis sine myositis) as a distinctive subset within the idiopathic inflammatory dermatomyopathies spectrum of clinical illness? PMID- 11907525 TI - The plight of descriptive dermatology and clinical investigation in an "evidence based" world. PMID- 11907526 TI - What to do with basal cell carcinoma leftovers. PMID- 11907527 TI - Lymphatic mapping and sentinel lymph node biopsy. PMID- 11907528 TI - The standard of care for Merkel cell carcinoma should include adjuvant radiation and lymph node surgery. PMID- 11907529 TI - Delayed bleomycin-induced hyperpigmentation and pressure on the skin. PMID- 11907530 TI - Basal cell carcinoma: shave biopsy versus punch biopsy technique in subtype diagnosis. PMID- 11907532 TI - Going green: Ireland. PMID- 11907533 TI - In search of sound science. PMID- 11907534 TI - Pursuing arrogant simplicities. PMID- 11907535 TI - Concern mounts as US agencies face challenges to data quality. PMID- 11907537 TI - Evolution critics seek role for unseen hand in education. PMID- 11907538 TI - Cash shortfall means time out for physicists. PMID- 11907539 TI - Breast-cancer survey sets screening age for women. PMID- 11907541 TI - US and Vietnam join forces to count cost of Agent Orange. PMID- 11907542 TI - Diplomats near pact in simmering debate over transgenic foods. PMID- 11907543 TI - Sea scouts plan big splash for oceanography. PMID- 11907546 TI - Harvard's melting pot. PMID- 11907547 TI - Peer review, unmasked. PMID- 11907548 TI - Biotech remains unloved by the more informed. PMID- 11907549 TI - Nothing automatic about ion-channel structures. PMID- 11907550 TI - Job-seekers, be careful of what you're signing. PMID- 11907551 TI - Opportunities for women in science (Russia, 1912). PMID- 11907557 TI - Biochemical ecology: how plants fight dirty. PMID- 11907558 TI - Planetary science: the north-south martian divide. PMID- 11907559 TI - Neurobiology: ready to unlearn. PMID- 11907561 TI - Molecular physiology: protecting the heart. PMID- 11907562 TI - Materials science: breaking the neural code. PMID- 11907563 TI - Quantum optics: light corralled. PMID- 11907564 TI - Immunology: the Wright stuff. PMID- 11907566 TI - Tumour biology: herceptin acts as an anti-angiogenic cocktail. AB - Malignant tumours secrete factors that enable them to commandeer their own blood supply (angiogenesis), and blocking the action of these factors can inhibit tumour growth. But because tumours may become resistant to treatments that target individual angiogenic factors by switching over to other angiogenic molecules, a cocktail of multiple anti-angiogenic agents should be more effective. Here we show that herceptin, a monoclonal antibody against the cell-surface receptor HER2 (for human epidermal growth factor receptor-2; ref. 4), induces normalization and regression of the vasculature in an experimental human breast tumour that overexpresses HER2 in mice, and that it works by modulating the effects of different pro- and anti-angiogenic factors. As a single agent that acts against multiple targets, herceptin, or drugs like it, may offer a simple alternative to combination anti-angiogenic treatments. PMID- 11907567 TI - Biochemistry: biosynthesis of an organofluorine molecule. AB - Although fluorine in the form of fluoride minerals is the most abundant halogen in the Earth's crust, only 12 naturally occurring organofluorine compounds have so far been found, and how these are biosynthesized remains a mystery. Here we describe an enzymatic reaction that occurs in the bacterium Streptomyces cattleya and which catalyses the conversion of fluoride ion and S-adenosylmethionine (SAM) to 5'-fluoro-5'-deoxyfluoroadenosine (5'-FDA). To our knowledge, this is the first fluorinase enzyme to be identified, a discovery that opens up a new biotechnological opportunity for the preparation of organofluorine compounds. PMID- 11907568 TI - The transorientation hypothesis for codon recognition during protein synthesis. AB - During decoding, a codon of messenger RNA is matched with its cognate aminoacyl transfer RNA and the amino acid carried by the tRNA is added to the growing protein chain. Here we propose a molecular mechanism for the decoding phase of translation: the transorientation hypothesis. The model incorporates a newly identified tRNA binding site and utilizes a flip between two tRNA anticodon loop structures, the 5'-stacked and the 3'-stacked conformations. The anticodon loop acts as a three-dimensional hinge permitting rotation of the tRNA about a relatively fixed codon-anticodon pair. This rotation, driven by a conformational change in elongation factor Tu involving GTP hydrolysis, transorients the incoming tRNA into the A site from the D site of initial binding and decoding, where it can be proofread and accommodated. The proposed mechanisms are compatible with the known structures, conformations and functions of the ribosome and its component parts including tRNAs and EF-Tu, in both the GTP and GDP states. PMID- 11907569 TI - Killing activity of neutrophils is mediated through activation of proteases by K+ flux. AB - According to the hitherto accepted view, neutrophils kill ingested microorganisms by subjecting them to high concentrations of highly toxic reactive oxygen species (ROS) and bringing about myeloperoxidase-catalysed halogenation. We show here that this simple scheme, which for many years has served as a satisfactory working hypothesis, is inadequate. We find that mice made deficient in neutrophil granule proteases but normal in respect of superoxide production and iodinating capacity, are unable to resist staphylococcal and candidal infections. We also show that activation provokes the influx of an enormous concentration of ROS into the endocytic vacuole. The resulting accumulation of anionic charge is compensated for by a surge of K+ ions that cross the membrane in a pH-dependent manner. The consequent rise in ionic strength engenders the release of cationic granule proteins, including elastase and cathepsin G, from the anionic sulphated proteoglycan matrix. We show that it is the proteases, thus activated, that are primarily responsible for the destruction of the bacteria. PMID- 11907570 TI - A topographically forced asymmetry in the martian circulation and climate. AB - Large seasonal and hemispheric asymmetries in the martian climate system are generally ascribed to variations in solar heating associated with orbital eccentricity. As the orbital elements slowly change (over a period of >104 years), characteristics of the climate such as dustiness and the vigour of atmospheric circulation are thought to vary, as should asymmetries in the climate (for example, the deposition of water ice at the northern versus the southern pole). Such orbitally driven climate change might be responsible for the observed layering in Mars' polar deposits by modulating deposition of dust and water ice. Most current theories assume that climate asymmetries completely reverse as the angular distance between equinox and perihelion changes by 180 degrees. Here we describe a major climate mechanism that will not precess in this way. We show that Mars' global north-south elevation difference forces a dominant southern summer Hadley circulation that is independent of perihelion timing. The Hadley circulation, a tropical overturning cell responsible for trade winds, largely controls interhemispheric transport of water and the bulk dustiness of the atmosphere. The topography therefore imprints a strong handedness on climate, with water ice and the active formation of polar layered deposits more likely in the north. PMID- 11907571 TI - Ferromagnetism in one-dimensional monatomic metal chains. AB - Two-dimensional systems, such as ultrathin epitaxial films and superlattices, display magnetic properties distinct from bulk materials. A challenging aim of current research in magnetism is to explore structures of still lower dimensionality. As the dimensionality of a physical system is reduced, magnetic ordering tends to decrease as fluctuations become relatively more important. Spin lattice models predict that an infinite one-dimensional linear chain with short range magnetic interactions spontaneously breaks up into segments with different orientation of the magnetization, thereby prohibiting long-range ferromagnetic order at a finite temperature. These models, however, do not take into account kinetic barriers to reaching equilibrium or interactions with the substrates that support the one-dimensional nanostructures. Here we demonstrate the existence of both short- and long-range ferromagnetic order for one-dimensional monatomic chains of Co constructed on a Pt substrate. We find evidence that the monatomic chains consist of thermally fluctuating segments of ferromagnetically coupled atoms which, below a threshold temperature, evolve into a ferromagnetic long range-ordered state owing to the presence of anisotropy barriers. The Co chains are characterized by large localized orbital moments and correspondingly large magnetic anisotropy energies compared to two-dimensional films and bulk Co. PMID- 11907572 TI - An ordered mesoporous organosilica hybrid material with a crystal-like wall structure. AB - Surfactant-mediated synthesis strategies are widely used to fabricate ordered mesoporous solids in the form of metal oxides, metals, carbon and hybrid organosilicas. These materials have amorphous pore walls, which could limit their practical utility. In the case of mesoporous metal oxides, efforts to crystallize the framework structure by thermal and hydrothermal treatments have resulted in crystallization of only a fraction of the pore walls. Here we report the surfactant-mediated synthesis of an ordered benzene-silica hybrid material; this material has an hexagonal array of mesopores with a lattice constant of 52.5 A, and crystal-like pore walls that exhibit structural periodicity with a spacing of 7.6 A along the channel direction. The periodic pore surface structure results from alternating hydrophilic and hydrophobic layers, composed of silica and benzene, respectively. We believe that this material is formed as a result of structure-directing interactions between the benzene-silica precursor molecules, and between the precursor molecules and the surfactants. We expect that other organosilicas and organo-metal oxides can be produced in a similar fashion, to yield a range of hierarchically ordered mesoporous solids with molecular-scale pore surface periodicity. PMID- 11907573 TI - Origin and fate of Lake Vostok water frozen to the base of the East Antarctic ice sheet. AB - The subglacial Lake Vostok may be a unique reservoir of genetic material and it may contain organisms with distinct adaptations, but it has yet to be explored directly. The lake and the overlying ice sheet are closely linked, as the ice sheet thickness drives the lake circulation, while melting and freezing at the ice-sheet base will control the flux of water, biota and sediment through the lake. Here we present a reconstruction of the ice flow trajectories for the Vostok core site, using ice-penetrating radar data and Global Positioning System (GPS) measurements of surface ice velocity. We find that the ice sheet has a significant along-lake flow component, persistent since the Last Glacial Maximum. The rates at which ice is frozen (accreted) to the base of the ice sheet are greatest at the shorelines, and the accreted ice layer is subsequently transported out of the lake. Using these new flow field and velocity measurements, we estimate the time for ice to traverse Lake Vostok to be 16,000 20,000 years. We infer that most Vostok ice analysed to date was accreted to the ice sheet close to the western shoreline, and is therefore not representative of open lake conditions. From the amount of accreted lake water we estimate to be exported along the southern shoreline, the lake water residence time is about 13,300 years. PMID- 11907574 TI - Development of anisotropic structure in the Earth's lower mantle by solid-state convection. AB - Seismological observations reveal highly anisotropic patches at the bottom of the Earth's lower mantle, whereas the bulk of the mantle has been observed to be largely isotropic. These patches have been interpreted to correspond to areas where subduction has taken place in the past or to areas where mantle plumes are upwelling, but the underlying cause for the anisotropy is unknown-both shape preferred orientation of elastically heterogeneous materials and lattice preferred orientation of a homogeneous material have been proposed. Both of these mechanisms imply that large-strain deformation occurs within the anisotropic regions, but the geodynamic implications of the mechanisms differ. Shape preferred orientation would imply the presence of large elastic (and hence chemical) heterogeneity whereas lattice-preferred orientation requires deformation at high stresses. Here we show, on the basis of numerical modelling incorporating mineral physics of elasticity and development of lattice-preferred orientation, that slab deformation in the deep lower mantle can account for the presence of strong anisotropy in the circum-Pacific region. In this model-where development of the mineral fabric (the alignment of mineral grains) is caused solely by solid-state deformation of chemically homogeneous mantle material anisotropy is caused by large-strain deformation at high stresses, due to the collision of subducted slabs with the core-mantle boundary. PMID- 11907575 TI - A ceratopsian dinosaur from China and the early evolution of Ceratopsia. AB - Ceratopsians (horned dinosaurs) represent one of the last and the most diverse radiations of non-avian dinosaurs. Although recent systematic work unanimously supports a basal division of Ceratopsia into parrot-like psittacosaurids and frilled neoceratopsians, the early evolution of the group remains poorly understood, mainly owing to its incomplete early fossil record. Here we describe a primitive ceratopsian from China. Cladistic analysis posits this new species as the most basal neoceratopsian. This new taxon demonstrates that some neoceratopsian characters evolved in a more incremental fashion than previously known and also implies mosaic evolution of characters early in ceratopsian history. PMID- 11907576 TI - Remains of Homo erectus from Bouri, Middle Awash, Ethiopia. AB - The genesis, evolution and fate of Homo erectus have been explored palaeontologically since the taxon's recognition in the late nineteenth century. Current debate is focused on whether early representatives from Kenya and Georgia should be classified as a separate ancestral species ('H. ergaster'), and whether H. erectus was an exclusively Asian species lineage that went extinct. Lack of resolution of these issues has obscured the place of H. erectus in human evolution. A hominid calvaria and postcranial remains recently recovered from the Dakanihylo Member of the Bouri Formation, Middle Awash, Ethiopia, bear directly on these issues. These approximately 1.0-million-year (Myr)-old Pleistocene sediments contain abundant early Acheulean stone tools and a diverse vertebrate fauna that indicates a predominantly savannah environment. Here we report that the 'Daka' calvaria's metric and morphological attributes centre it firmly within H. erectus. Daka's resemblance to Asian counterparts indicates that the early African and Eurasian fossil hominids represent demes of a widespread palaeospecies. Daka's anatomical intermediacy between earlier and later African fossils provides evidence of evolutionary change. Its temporal and geographic position indicates that African H. erectus was the ancestor of Homo sapiens. PMID- 11907577 TI - Genetic cost of reproductive assurance in a self-fertilizing plant. AB - The transition from outcrossing to self-fertilization is one of the most common evolutionary trends in plants. Reproductive assurance, where self-fertilization ensures seed production when pollinators and/or potential mates are scarce, is the most long-standing and most widely accepted explanation for the evolution of selfing, but there have been few experimental tests of this hypothesis. Moreover, many apparently adaptive floral mechanisms that ensure the autonomous production of selfed seed might use ovules that would have otherwise been outcrossed. This seed discounting is costly if selfed offspring are less viable than their outcrossed counterparts, as often happens. The fertility benefit of reproductive assurance has never been examined in the light of seed discounting. Here we combine experimental measures of reproductive assurance with marker-gene estimates of self-fertilization, seed discounting and inbreeding depression to show that, during 2 years in 10 Ontario populations of Aquilegia canadensis (Ranunculaceae), reproductive assurance through self-fertilization increases seed production, but this benefit is greatly outweighed by severe seed discounting and inbreeding depression. PMID- 11907578 TI - Reduced adaptation of a non-recombining neo-Y chromosome. AB - Sex chromosomes are generally believed to have descended from a pair of homologous autosomes. Suppression of recombination between the ancestral sex chromosomes led to the genetic degeneration of the Y chromosome. In response, the X chromosome may become dosage-compensated. Most proposed mechanisms for the degeneration of Y chromosomes involve the rapid fixation of deleterious mutations on the Y. Alternatively, Y-chromosome degeneration might be a response to a slower rate of adaptive evolution, caused by its lack of recombination. Here we report patterns of DNA polymorphism and divergence at four genes located on the neo-sex chromosomes of Drosophila miranda. We show that a higher rate of protein sequence evolution of the neo-X-linked copy of Cyclin B relative to the neo-Y copy is driven by positive selection, which is consistent with the adaptive hypothesis for the evolution of the Y chromosome. In contrast, the neo-Y-linked copies of even-skipped and roundabout show an elevated rate of protein evolution relative to their neo-X homologues, probably reflecting the reduced effectiveness of selection against deleterious mutations in a non-recombining genome. Our results provide evidence for the importance of sexual recombination for increasing and maintaining the level of adaptation of a population. PMID- 11907579 TI - Dissecting the architecture of a quantitative trait locus in yeast. AB - Most phenotypic diversity in natural populations is characterized by differences in degree rather than in kind. Identification of the actual genes underlying these quantitative traits has proved difficult. As a result, little is known about their genetic architecture. The failures are thought to be due to the different contributions of many underlying genes to the phenotype and the ability of different combinations of genes and environmental factors to produce similar phenotypes. This study combined genome-wide mapping and a new genetic technique named reciprocal-hemizygosity analysis to achieve the complete dissection of a quantitative trait locus (QTL) in Saccharomyces cerevisiae. A QTL architecture was uncovered that was more complex than expected. Functional linkages both in cis and in trans were found between three tightly linked quantitative trait genes that are neither necessary nor sufficient in isolation. This arrangement of alleles explains heterosis (hybrid vigour), the increased fitness of the heterozygote compared with homozygotes. It also demonstrates a deficiency in current approaches to QTL dissection with implications extending to traits in other organisms, including human genetic diseases. PMID- 11907580 TI - Inhibition of climbing fibres is a signal for the extinction of conditioned eyelid responses. AB - A fundamental tenet of cerebellar learning theories asserts that climbing fibre afferents from the inferior olive provide a teaching signal that promotes the gradual adaptation of movements. Data from several forms of motor learning provide support for this tenet. In pavlovian eyelid conditioning, for example, where a tone is repeatedly paired with a reinforcing unconditioned stimulus like periorbital stimulation, the unconditioned stimulus promotes acquisition of conditioned eyelid responses by activating climbing fibres. Climbing fibre activity elicited by an unconditioned stimulus is inhibited during the expression of conditioned responses-consistent with the inhibitory projection from the cerebellum to inferior olive. Here, we show that inhibition of climbing fibres serves as a teaching signal for extinction, where learning not to respond is signalled by presenting a tone without the unconditioned stimulus. We used reversible infusion of synaptic receptor antagonists to show that blocking inhibitory input to the climbing fibres prevents extinction of the conditioned response, whereas blocking excitatory input induces extinction. These results, combined with analysis of climbing fibre activity in a computer simulation of the cerebellar-olivary system, suggest that transient inhibition of climbing fibres below their background level is the signal that drives extinction. PMID- 11907581 TI - Oestrogen protects FKBP12.6 null mice from cardiac hypertrophy. AB - FK506 binding proteins 12 and 12.6 (FKBP12 and FKBP12.6) are intracellular receptors for the immunosuppressant drug FK506 (ref. 1). The skeletal muscle ryanodine receptor (RyR1) is isolated as a hetero-oligomer with FKBP12 (ref. 2), whereas the cardiac ryanodine receptor (RyR2) more selectively associates with FKBP12.6 (refs 3, 4, 5). FKBP12 modulates Ca2+ release from the sarcoplasmic reticulum in skeletal muscle and developmental cardiac defects have been reported in FKBP12-deficient mice, but the role of FKBP12.6 in cardiac excitation contraction coupling remains unclear. Here we show that disruption of the FKBP12.6 gene in mice results in cardiac hypertrophy in male mice, but not in females. Female hearts are normal, despite the fact that male and female knockout mice display similar dysregulation of Ca2+ release, seen as increases in the amplitude and duration of Ca2+ sparks and calcium-induced calcium release gain. Female FKBP12.6-null mice treated with tamoxifen, an oestrogen receptor antagonist, develop cardiac hypertrophy similar to that of male mice. We conclude that FKBP12.6 modulates cardiac excitation-contraction coupling and that oestrogen plays a protective role in the hypertrophic response of the heart to Ca2+ dysregulation. PMID- 11907582 TI - Nitric oxide regulates the heart by spatial confinement of nitric oxide synthase isoforms. AB - Subcellular localization of nitric oxide (NO) synthases with effector molecules is an important regulatory mechanism for NO signalling. In the heart, NO inhibits L-type Ca2+ channels but stimulates sarcoplasmic reticulum (SR) Ca2+ release, leading to variable effects on myocardial contractility. Here we show that spatial confinement of specific NO synthase isoforms regulates this process. Endothelial NO synthase (NOS3) localizes to caveolae, where compartmentalization with beta-adrenergic receptors and L-type Ca2+ channels allows NO to inhibit beta adrenergic-induced inotropy. Neuronal NO synthase (NOS1), however, is targeted to cardiac SR. NO stimulation of SR Ca2+ release via the ryanodine receptor (RyR) in vitro, suggests that NOS1 has an opposite, facilitative effect on contractility. We demonstrate that NOS1-deficient mice have suppressed inotropic response, whereas NOS3-deficient mice have enhanced contractility, owing to corresponding changes in SR Ca2+ release. Both NOS1-/- and NOS3-/- mice develop age-related hypertrophy, although only NOS3-/- mice are hypertensive. NOS1/3-/- double knockout mice have suppressed beta-adrenergic responses and an additive phenotype of marked ventricular remodelling. Thus, NOS1 and NOS3 mediate independent, and in some cases opposite, effects on cardiac structure and function. PMID- 11907583 TI - TNF-RII and c-IAP1 mediate ubiquitination and degradation of TRAF2. AB - Tumour necrosis factor-alpha (TNF-alpha) is a proinflammatory mediator that exerts its biological functions by binding two TNF receptors (TNF-RI and TNF RII), which initiate biological responses by interacting with adaptor and signalling proteins. Among the signalling components that associate with TNF receptors are members of the TNF-R-associated factor (TRAF) family. TRAF2 is required for TNF-alpha-mediated activation of c-Jun N-terminal kinase (JNK), contributes to activation of NF-kappaB, and mediates anti-apoptotic signals,. TNF RI and TNF-RII signalling complexes also contain the anti-apoptotic ('inhibitor of apoptosis') molecules c-IAP1 and c-IAP2 (refs 5, 6), which also have RING domain-dependent ubiquitin protein ligase (E3) activity. The function of IAPs in TNF-R signalling is unknown. Here we show that binding of TNF-alpha to TNF-RII induces ubiquitination and proteasomal degradation of TRAF2. Although c-IAP1 bound TRAF2 and TRAF1 in vitro, it ubiquitinated only TRAF2. Expression of wild type c-IAP1, but not an E3-defective mutant, resulted in TRAF2 ubiquitination and degradation. Moreover, E3-defective c-IAP1 prevented TNF-alpha-induced TRAF2 degradation and inhibited apoptosis. These findings identify a physiologic role for c-IAP1 and define a mechanism by which TNF-RII-regulated ubiquitin protein ligase activity can potentiate TNF-induced apoptosis. PMID- 11907586 TI - [Autism:diagnosis criteria. Cognitive-evolutionary approach]. AB - OBJECTIVE: To get acquainted with the information handled by health professionals regarding autism and its diagnosis criteria in the province of Mendoza, Argentina. METHODOLOGY: Qualitative. A descriptive and exploratory study was conducted on a non-probabilistic sample of 38 professionals of Mendoza province. The professionals answered a questionnaire on autism and its diagnosis criteria. RESULTS: Fifty-five percent of the professionals of the sample did not diagnose autism. The professionals that did (45percent of the sample), though updated as regards diagnosis criteria, did not handle actual information about origin and type of this pathology, its symptoms and characteristics. Since most professionals belong to public institutions of the province, these results become relevant. PMID- 11907587 TI - [Phenomena of daily life and mental pathology]. AB - To get acquainted with the information handled by health There are many psychic phenomena that may be observed both among diagnosticated patients and among persons included in the statistical media of normality. Reality as continuum, perception and some particular forms of it (pareidolias, sensitive pseudo-images, hipnogogical and hipnopompical hallucinations, eidetic images, sensitive post images, dej racont, cout, vu, etc.) as well as premonitions, epiphanies, mystical experiences and overvalued, delirious, encapsulated, fixed ideas, would be characteristic of mental state depending on their intensity and their place in the psychic life. These events are assessed according society and the general circumstances in each situation, and they are presented seperately only by classificatory interest. PMID- 11907588 TI - [Implications of cAMP signaling in affective disorders]. AB - The last decade brought about a shift in the theoretical framework, addressing the issue of affective disorders. During this period, the research endeavors have been directed towards investigating the biochemical mechanisms involved in the transduction of informations from the self surface to the cell interior. Nowadays, the emerging picture is that various signal transduction pathways could be linked to the pharmacotherapy and pathophysiology of affective disorders. Among these pathways, in this review the role of the cAMP signaling system will be highlighted. In particular, a summary of the preclinical and clinical data suggesting the involvement of the downstream components of this system in affective disorders will be given. This framework has the potential to improve our knowledge providing clues for the development of novel research strategies in the field of affective disorders. PMID- 11907589 TI - [The cost of bipolar disorder: our situation]. AB - Recent studies are showing that prevalence of Bipolar Disorders is higher than traditionally admitted, and that their costs are equal, at least, to those arising from schizophrenia. In our region, these disorders may be considered as an epidemic in the few next years, according the epidemiological approach. Nevertheless, the very few available data in our country show a serious deficit in diagnosis, treatment and information available for the users. In this context, the inertia of government is critical in generating specific policies aimed to control these disorders. Facing this challenge, needed information should be achieved in order to be able of establishing rational policies and strategies thus increasing general information about this problem, involving the users. PMID- 11907590 TI - [Update on the pharmacological treatment of bipolar disorder]. AB - The main goal of the treatment of Bipolar Disorder is to achieve and to maintain the euthymia by using mood stabilizing drugs. This article reviews the available pharmacological treatments of Bipolar Disorder, emphazising not pharmacology but its effectiveness in mania, bipolar depression, mixed states and rapid cyclers. PMID- 11907591 TI - [Mixed states]. AB - The clinical features of mixed states and their variants are reviewed, emphasizing the diagnostic versatility, dynamics and differences as compared to others pathologies. Different classificatory views are also reviewed, ranging from the more restrictive to the broader ones. PMID- 11907592 TI - [Psychosocial therapy of bipolar disorder]. AB - Pharmacotherapy is the treatment of choice for Bipolar Disorder without any doubt. However, an integration of psychotherapeutic techniques with pharmacotherapy has been recommended by the American Psychiatric Association. Among the different psychosocial approaches we shall quote the most used ones. Specially psychoeducation, which is considered one of the most useful ways. We shall report our experience working with the psychoeducational method in the treatment of the Bipolar Disorder during last years. PMID- 11907594 TI - [Psychotherapy of bipolar disorder]. AB - Nowadays, it exists evidence that Bipolar Disorder have a less favourable forecast that it was thought. However, there are drugs that prevent recurrences and they dominate therapeutic management. Psychosocial disturbs that are caused by that Disorder make necessary psycotherapeutic interventions. That article includes a description of three psychotherapies (Group Psychotherapy, Family Psychotherapy and Interpersonal Psychotherapy) addressed to this group of patients. PMID- 11907593 TI - [Some concepts about the treatment of the depressive episode in bipolar disorder]. AB - A frequent clinical mistake is to diagnose a Depressive Episode of the Bipolar Disorder as a Unipolar Depression. This, generally, has iatrogenic consequences for the patient. In this article the approaches are revised for a correct diagnosis and also an algorithm for an appropriate treatment is suggested. PMID- 11907595 TI - [Bipolar disorder. 1854]. PMID- 11907596 TI - [Psychotherapy in prepaid health plans]. PMID- 11907598 TI - [Clinical medicine and administration, or how to survive managed care]. PMID- 11907599 TI - [Psychiatric comorbidity in clinical Tourette Syndrome's samples]. AB - Classically conceived essentially as a rare neurological disorder, Tourette Syndrome (TS) is frequently associated with comorbid disorders such as attention deficit, obsessive-compulsive disorder, impulse control problem and a variety of other behavioral symptoms that can clearly have an impact on cognitive,educational and psychosocial function and can be more debilitating than the cardinal motor features. The full spectrum of TS is often not recognized in our medical community while it's usual atributtion to exclusive pshycogenic causes leads to underdiagnosis. Meanwhile epidemiological studies shows it's increasingly prevalence and its pottencially multiple deficits, specially on pediatrical clinical population. Through her experience as a tourette's clinical researcher the author review Tourette syndrome's clinical features and it's natural course, sharing the results and conclusions of an interdisciplinary clinical research about The Importance of an Early Diagnosis of Comorbidity in a Clinical Sample of 51 argentine patients with TS. PMID- 11907600 TI - [Empirical psychotherapy research in Argentina. Current situation, methods and problems]. AB - This article presents a brief report about empirical psychotherapy research in Argentina. Specifically, it analyzes some of the problems related to doing psychological treatments research in our country, beyond the theoretical background and preferences associated to the research. This article also gives an overview on some of the psychotherapy research techniques that are being used in the mentioned environment. For this study a total of 26 on going works were analyzed, to identify some of its main characteristics, and to define the key themes and research procedures used in Argentina. In order to exemplify the characteristics of the most used methods in this area, a summary describing its aims and procedures is included. PMID- 11907601 TI - [Empirical psychotherapy research]. AB - Current bibliography on epistemological status of psychoanalysis is reviewed. In particular, claims from several fields of knowledge, including those from "hard" sciences as physics, mathematics or philosophy of sciencies about empirical demonstration of efficacy and theoretical basis of the discipline are discussed. PMID- 11907602 TI - [Clinical inferential process in psychoanalysis]. AB - OBJECTIVE: Despite the highest importance of studying the inferential process in psychotherapy, few systematic studies deal with the ways in which psychotherapists work with the material offered by their patients and construct clinical hypotheses about it. The purpose of this study is to present empirical findings about similarities and differences in the way psychoanalysts from different theorethical orientations and level of clinical experience produce their clinical inferences. METHODOLOGY: The sample includes 20 psychoanalysts selected according to their theoretical orientation (Freudian and Lacanian psychoanalysts) and their level of clinical experience (less than ten years and more than twenty years of clinical practice). They listened to the same tape recorded first session of psychotherapy and were asked to report their clinical hypotheses, clues, and hunches about the material. They also answered a questionnaire about their inferential work. RESULTS: Freudian psychoanalysts produced more single hypothesis and the same number of combined hypothesis than lacanian psychoanalysts. The level of experience was a factor when it came to producing combined hypothesis, no matter their theoretical framework. 90 per cent of the interviewed made their first clinical inference before the first 9 minutes of the session (regardless of their experience and framework), and agreed on the core themes of the clinical case. CONCLUSIONS: there are similarities and differences in the inferential work according to the theoretical orientation and clinical experience of the psychoanalysts. Their level of experience has a positive matching with their capacity to produce more combined inferences. Most of the clinical inferences are psychological and relational, and they are produced at the very beginning of the session. PMID- 11907603 TI - [Psychotherapy reasearch: a bridge between theory and clinic]. AB - This paper presents a panorama of current developments in psychotherapy research. Starting by an historical brief review of the evolution of this discipline, the two main streams in which this field is nowadays divided are examined. The first one, represented by those who favor the question of efficacy, promotes the development of empirically validated treatments (EVT) based on experimental studies. Conversely, the other one, represented by those who favor effectiveness, argue for the advisability of following a generalizability criteria that attempts to examine the results of psychotherapy in naturalistic contexts. PMID- 11907604 TI - [Therapeutic alliance and analytic setting]. AB - OBJECTIVE: The goal of this work is to study the relationship between the therapeutic alliance, the subjective perception of improvement, the frequency of sessions and the type of analytic interventions, in both psychoanalysts and non psychoanalysts patients. METHODOLOGY: 39 subjects under psychoanalytic treatment lasting one to six years (mean 4.2 years) were interviewed. It was performed: a) a therapeutic alliance evaluation scale (HRQ); b) a subjetive improvement perception scale (PSM); c) a scale to evaluate the style of the psychoanalytic interventions (EI). The sample was divided in two groups: 1) 18 non psychoanalysts under psychoanalytic treatment, who assited to a mean of 1.15 sessions per week (group 1) and b) 21 psychoanalysts receiving two kinds of psychoanalytic treatments: a) one following the international Psychoanalytc Associations rules (group 2A), b) the other with 1.65 mean sessions per week (group 2B). RESULTS: a) patients in groups 1 and 2A showed similar HRQ scores, and both were higher than that shown by group 2B (21.53 vs 21.51 vs 17.22) No differences were found neither in PSM scores (3.61 vs 3.85 vs 3.85 respectively) nor in the EI scores (3.61 vs 3.71 vs 3.71). It was observed a positive correlation between HRQ and PSM (group 1: r: 0.55 and gorup 2, r: 0.31) but no correlation was found neither with the number of sessions per week (group 1, r:0.13; group 2, r: 0.30) nor with EI score (group 1, r: -0.21; group 2, r: 0.08). DISCUSION AND CONCLUSIONS: a) intensity of perceived therapeutic alliance is correlated with improvement but b) is not correlated with sessions frequency or style of psychoanalytic interventions. It is also discussed which psychic changes are related with the therapeutic alliance with regards with different psychoanalytic theoretical frames. PMID- 11907605 TI - [Phsychotherapy results: What do our patients think?]. AB - The present paper shows the way a mental health organization works and, at the same time, examines some results upon the post-therapy follow-up for outpatient treatments, the therapists' qualities perceived by the patients and the readmission rate. The results are twofold: for internal purposes, they allow a feedback for continuous quality improvement. For the outside, they generate information for future comparisons among other similar organizations. This paper also address the debate between two major stands in the field of psychotherapy research (one for the Evidence Based Psychotherapy and the other for the Psychotherapy Based on Common Features). PMID- 11907607 TI - The Canadian Institutes for Health Research and stem cell guidelines. PMID- 11907608 TI - Nutrition tools for patients: helping patients making healthful choices! PMID- 11907609 TI - Decline in ischemic heart disease mortality. PMID- 11907611 TI - A wake-up call. PMID- 11907612 TI - Canadian Cardiovascular Congress address: why no one in government listens to doctors. PMID- 11907613 TI - The number remaining at risk: an adjunct to the number needed to treat. AB - Although the number of patients needed to treat (NNT) to prevent an adverse clinical event is of great clinical value to practising physicians, it is limited in that it fails to provide a measure of prognosis among patients not achieving benefit. For example, if the NNT is 100, what is likely to happen to the other 99? The number remaining at risk (NRR), which is an index that enhances the value of the NNT, is described. The NRR is the ratio of the residual event rate among treated patients and the absolute reduction in outcome events (NRR = experimental event rate [EER]/control event rate [CER]-EER), where EER and CER are the event rates among experimental and control groups, respectively. This index represents the number of events likely to occur among the NNT, or the odds of experiencing an adverse outcome event as opposed to deriving benefit from therapy. The NRR is a simple index that can easily be calculated from the published results of a clinical trial. As an adjunct to the NNT, it provides a measure of the impact of therapy and the average prognosis of remaining patients. PMID- 11907614 TI - Validation of an automated method for assessing brachial artery endothelial dysfunction. AB - BACKGROUND: Endothelial dysfunction is an early finding in diverse vascular diseases and can be measured noninvasively using brachial ultrasound. There is great interest in the potential use of this parameter for assessing interventions or cardiovascular prognosis. Automated image analysis of the ultrasound images would facilitate implementation of such measurements in high throughput clinics and/or large clinical trials. OBJECTIVES: To compare a new method designed to assess brachial artery diameter and percentage diameter changes through automated, beat-by-beat image processing (Brachial Tools, Medical Imaging Applications, USA), with a nonautomated method (Prosound System, Jet Propulsion Laboratory, USA). PATIENTS AND METHODS: Brachial ultrasound tapes from 12 patients undergoing endothelial function assessment using forearm cuff-occlusion, measurement of flow-mediated dilation and responses to nitroglycerin were analyzed by both methods. RESULTS: The correlation between the two systems was excellent for both the measurement of absolute diameters (r=0.995, P<0.001) and percentage diameter changes (r=0.973, P<0.001). The automated method demonstrated no bias compared with the frame-by-frame method and excellent precision (0.07 mm and 1.62 percentage diameter change). CONCLUSIONS: The automated method provides valid data while substantially diminishing analysis time. PMID- 11907615 TI - Thrombus formation on guide wires during routine PTCA procedures: a scanning electron microscopic evaluation. AB - BACKGROUND: Various coatings are used on catheters and guide wires to improve resistance to surface thrombus formation. However, little is known about the thrombogenicity of 0.014" angioplasty guide wires and the protection offered by these coatings. OBJECTIVES: To evaluate the thrombogenicity of five different guide wires used in coronary angioplasty. METHODS: Five different 0.014" guide wire types were evaluated in 50 consecutive angioplasty procedures. At the end of the procedure, the distal part of the guide wire was cut, put in formalin and prepared for scanning electron microscopic evaluation. The condition of each guide wire was then classified into one of three predefined categories: no thrombus, limited thrombus and significant thrombus formation. RESULTS: Silicone (n=10, Guidant, USA), phosphorylcholine polymer (n=8, Biocompatibles, Ireland), hydrophilic polymer (n=8, Scimed, USA) and two Teflon-based coatings (n=16, Schneider, USA; n=8, Cordis, USA) were evaluated. On microscopic examination, 48% of guide wires had a significant amount of thrombus, 18% had limited thrombus formation and 34% had no thrombus. The results were very dissimilar among the groups. Significant thrombus was found on 80% of Guidant guide wires, 69% of Schneider guide wires, 38% of Scimed guide wires and 25% of Cordis guide wires, while none was found on Biocompatibles guide wires (P<0.0001). CONCLUSIONS: Significant thrombus formation on angioplasty guide wires was a frequent finding, occurring in 48% of cases. Resistance to thrombus formation was very dissimilar among coatings, with only the Biocompatibles phosphorylcholine-coated guide wires showing no thrombus formation at all. Whether subclinical thromboembolization occurred in some patients is unknown, and the clinical implications of this study remain to be defined. PMID- 11907616 TI - Effects of plaque composition on vascular remodelling after angioplasty in the MultiVitamins and Probucol (MVP) trial. AB - BACKGROUND: The antioxidant probucol reduced coronary restenosis in the MultiVitamins and Probucol (MVP) trial by improving vascular remodelling. Whether calcification limits the extent of adaptive vessel enlargement is not known. OBJECTIVE: To determine whether plaque composition at the dilated site affects probucol-induced vascular remodelling after angioplasty. PATIENTS AND METHODS: Beginning 30 days before percutaneous transluminal coronary angioplasty (PTCA), 317 patients received either probucol, vitamins, probucol and vitamins, or placebo. Patients were then treated for six months after PTCA. Intravascular ultrasound (IVUS) was performed post-PTCA and at follow-up in 94 patients (111 segments). The cross-section for serial analysis was the one at the angioplasty site with the smallest lumen area at follow-up. Quantitative analysis consisted of measurements of lumen area and external elastic membrane (EEM) area. The selected cross-section was also divided into five regions according to the type of plaque present (calcific, fibrotic, hypoechoic, fibrohypoechoic or normal). Plaque characterization scores (PCS) (PCS for arc, area, inner perimeter and outer perimeter) were calculated using weighting factors. RESULTS: There were no interactions between potential PCS covariates and probucol main effect on changes in lumenal, EEM and wall area. There were no significant PCS covariates in the model for change in EEM as they were all removed using a backward stepwise procedure. The last potential covariate (area PCS) had a significance level of P=0.48. In contrast, probucol significantly influenced the change in EEM over time (P=0.003). CONCLUSION: Plaque composition at the dilated site does not appear to influence probucol-induced vascular remodelling after angioplasty. PMID- 11907618 TI - Diagnostic endomyocardial biopsy pathology: secondary myocardial diseases and other clinical indications - a review. AB - Endomyocardial biopsy is a commonly performed, useful procedure for the evaluation of cardiac tissues for many indications, including the assessment of transplant rejection, myocarditis, cardiomyopathy, drug toxicity, neoplastic involvement, chest pain, arrhythmia and secondary involvement by systemic diseases. The present paper focuses on clinicopathological considerations for biopsy use for individuals with secondary myocardial disease. The use of endomyocardial biopsy for the clinical indications of idiopathic arrhythmia and chest pain is also discussed. PMID- 11907617 TI - Dietary fish oil does not prevent doxorubicin-induced cardiomyopathy in rats. AB - BACKGROUND: Dietary fish oil potentiates the susceptibility of cellular membranes to lipid peroxidation, although it is also known to have beneficial effects on the development of cardiovascular diseases. OBJECTIVE: The effects of dietary fish oil against doxorubicin-induced cardiomyopathy, in which free radicals and lipid peroxidation are involved, were investigated in rats. ANIMALS AND METHODS: Sprague-Dawley rats (100 g) were fed a standard diet or a high fish oil diet (containing 10% fish oil) throughout the experimental period. Four weeks after starting each diet, experimental rats were treated with doxorubicin (cumulative dose 15 mg/kg) or vehicle (0.28 M dextrose solution). After three weeks of doxorubicin treatment, the cardiac performance, myocardial lipid peroxidation and myocardial vitamin E level were assessed. RESULTS: Compared with control rats, doxorubicin-treated rats showed a significantly increased mortality rate (P<0.05), and significantly decreased systolic blood pressure and left ventricular fractional shortening (P<0.01). The myocardial thiobarbituric acid reactive substance level was significantly higher in doxorubicin-treated rats than in control rats (P<0.01), while the myocardial vitamin E level was significantly lower (P<0.05). Dietary fish oil enhanced the myocardial lipid peroxidation caused by doxorubicin, which was associated with a further decrease in myocardial vitamin E level. As a result, the rats treated with both doxorubicin and the high fish oil diet showed the highest mortality rate and the lowest cardiac performance of all the experimental groups. CONCLUSIONS: Dietary fish oil may reduce antioxidant defences and accelerate susceptibility of the myocardium to lipid peroxidation in rats under doxorubicin treatment. This may partly explain why dietary fish oil does not prevent doxorubicin-induced cardiomyopathy. PMID- 11907619 TI - Single chamber ventricular compared with dual chamber pacing: a review. AB - Since its introduction in the 1950s, the cardiac pacemaker has become increasingly sophisticated in an attempt to mimic normal cardiac physiology. Rapidly evolving pacing technology has seen pacemakers evolve from crude, fixed rate, single chamber ventricular devices to dual chamber rate-adaptive units. While there is indirect evidence that supports the use of dual chamber pacing in the vast majority of patients, it is still unclear whether these newer, more expensive devices afford a significant morbidity and mortality benefit over single-chamber, ventricular, rate-adaptive pacemakers. A review of three large, randomized trials failed to demonstrate a clear benefit of dual chamber or atrial based pacing over single chamber ventricular pacing for the majority of cardiovascular outcomes (heart failure, stroke and mortality), with the possible exception of atrial fibrillation. Information is also needed on the potential protective effects of atrial-based pacing over dual chamber pacing in elderly patients with sinus node dysfunction. Longer follow-up periods may be necessary to determine whether there are any mortality benefits associated with dual chamber pacing. Additional confirmation of benefits of dual chamber pacing may be provided by other ongoing prospective trials. PMID- 11907621 TI - Fairness and the media. PMID- 11907620 TI - Images in Cardiology. Postaortotomy false aneurysm of the ascending aorta. PMID- 11907622 TI - Autosomal aberrations associated with testicular dysgenesis or spermatogenic arrest in Chinese patients. AB - AIM: To analyze the relationship between autosomal aberrations and testicular dysgenesis or spermatogenic arrest in Chinese patients and to map the corresponding regions on each autosome in regard to the recorded aberrations accompanying these distubances. METHODS: One hundred and nineteen cases of aberrant karyotypes with testicular dysgenesis, azoospermia or oligozoospermia reported in five Chinese journals and one monograph were analyzed. For each autosome, the type and frequency of chromosomal aberrations were counted and the regions corresponding to the disturbances were mapped out. RESULTS: Chromosomes 13, 14, 9, 21 exhibited a high frequency of aberration and bands 14q11 and 13p11 were the two regions showing the highest linkage to testicular dysgenesis or infertility. The frequency of chromosomal aberrations was higher in bands 9p11 and 22q than in others. CONCLUSION: Autosomes 13, 14, 9 and 21 in the order of importance play a critical role in testicular development and spermatogenesis and other autosomes may also contribute; the following regions, 14q11, 13p11,9p11, and 22q, are of high significance. PMID- 11907623 TI - Testicular sperm extraction in azoospermic patients with gonadal germ cell tumors prior to chemotherapy--a new therapy option. AB - BACKGROUND: In view of the high cure rates in patients with testicular germ cell tumors (TGCT), increasing clinical importance is attached to protection of fertility. Long-term infertility due to cytostatic therapy may be expected in more than 50% of the patients at a cumulative dose of cisplatin > 0.6 g/m2. The standard procedure for fertility protection in cancer patients includes cryopreservation of ejaculated spermatozoa. Considering that some patients have tumor-induced azoospermia, we examined the usefulness of testicular sperm extraction before therapy. METHOD: A survey of the literature served as a basis for investigating biological and clinical aspects of the impact of chemotherapy on male fertility. A study of our patient population also enabled us to explore the option of extracting sperm from the contralateral healthy testis prior to treatment in 14 azoospermic patients with testicular germ cell tumors. RESULTS: We were able to successfully recover haploid germ cells in 6/14 testicular biopsies from azoospermic patients with testicular germ cell cancer prior to treatment. Maturation arrest was found in 3/14 cases and Sertoli-cell-only syndrome in the rest. None of the patients had secondary healing or a treatment delay because of the testicular biopsy. CONCLUSION: Since the post-therapeutic fertility status is difficult to predict in cancer patients, we think that TESE should be regarded as a general option prior to cancer treatment and offered to azoospermic cancer patients. New guidelines should be established in this connection. PMID- 11907624 TI - Chloroform extract of Carica papaya seeds induces long-term reversible azoospermia in langur monkey. AB - AIM: To evaluate the antifertility activity of the chloroform extract of Carica papaya seeds by oral administration in langur monkey, Presbytis entellus entellus. METHODS: The chloroform extract of Carica papaya seeds, 50 mg/kg/day, was administered orally for 360 days to adult male langur monkeys. The sperm characteristics by light and electron microscopy, the sperm functional tests, the semen biochemistry, the serum testosterone level, the Leydig cell function, and the histology and ultrastructure of testis were determined to evaluate the antifertility activity and the blood biochemistry and hematology, to evaluate the toxicology. RESULTS: The extract gradually decreased the sperm concentration since days 30-60 of treatment with a total inhibition of sperm motility, a decrease in sperm viability and increase in sperm abnormality. Azoospermia was observed after day 90 of treatment and continued during the whole treatment period. Treatment withdrawal resulted in a gradual recovery in these parameters and 150 days later they reverted to nearly the pretreatment values. Morphological observation of the ejaculated sperm by light and scanning electron microscopy showed deleterious changes, particularly on the mid-piece. Sperm functional tests, viz., sperm mitochondrial activity index, acrosome intactness test and hypo-osmotic swelling test scored in the infertile range during treatment and returned to the fertile values 150 days after drug withdrawal. Histology of the testis revealed shrunken tubules, germ cell atrophy and normal Leydig cells. Ultrastructure of the testis showed vacuolization in the cytoplasm of Sertoli cells and germ cells. Loss of cytoplasmic organelles were evident in the spermatocytes and spermatids. Round spermatids showed loss of Golgi bodies, peripheral mitochondria and vacuolated cytoplasm, indicating maturational arrest. Leydig cell functional test indicated a mild inhibition of steroidogenic function. Haematology and serum biochemistry study disclosed no significant toxicological effect and the serum testosterone level was not affected. CONCLUSION: Carica papaya seed extract may selectively act on the developing germ cells, possibly mediated via Sertoli cells, leading to azoospermia. PMID- 11907625 TI - N-acetyl beta-D-glucosaminidase is not attached to human sperm membranes through the glycosylphosphatidyl inositol (GPI)-anchor. AB - AIM: The mode of anchorage of N-acetyl beta-D-glucosaminidase (NAGA) on human ejaculated sperm was investigated. METHODS: Sperm plasma membrane was prepared by discontinuous sucrose gradient centrifugation from human sperm. NAGA was solubilized from these membranes by two detergents: octyl-glycoside and triton X 100. In separate studies, the release of the enzyme from the sperm membrane preparation by phosphatidylinositol specific phospholipase C (PI-PLC) was also examined. RESULTS: NAGA activity was detected on sperm membranes isolated from human ejaculates. The pattern of the enzyme solubilization by detergents indicated that the enzyme was an integral protein of sperm membrane. NAGA was not released from the sperm membranes by PI-PLC treatment. CONCLUSION: The evidence presented strongly suggests that human sperm membrane bound NAGA is not attached via the GPI anchor. PMID- 11907626 TI - Effects of pyrethroid insecticide ICON (lambda cyhalothrin) on reproductive competence of male rats. AB - AIM: To assess the effect of ICON (trade name of lambda-cyhalothrin) on sexual competence and fertility of male rats. METHODS: Male rats were gavaged daily for 7 consecutive days with different doses of ICON (63 mg/kg and 100 mg/kg) or vehicle (distilled water). Their sexual behaviour and fertility were evaluated at different time points during treatment and post-treatment using receptive females. RESULTS: Treatment had no effect on fertility, but sexual competence was seriously impaired: libido (assessed in terms of pre-coital sexual behaviour, and numbers of mounting, intromission and ejaculation), sexual arousability/motivation (in terms of latencies for mounting, intromission and ejaculation), sexual vigour (judged by frequencies of mounting and intromission or copulatory efficiency). In addition, ICON suppressed intromission ratio, indicating erectile dysfunction. These effects on sexual function had a rapid onset and was reversible. ICON-induced sexual dysfunction was mediated by multiple mechanisms, mainly toxicity, stress, sedation and possibly via GABA and dopaminergic systems. CONCLUSION: Exposure to ICON may cause sexual dysfunction in male rats. PMID- 11907628 TI - Morphometric study on leydig cells in capsulotomized testis of rats. AB - AIM: To further clarify the changes occurred in the testicular capsulotomized rats. METHODS: In testicular capsulotomized and sham-operated rats, the cross sectional area, the nucleus diameter and the number of Leydig cells were morphologically analyzed by the Vidas Image Processing System connected to a microscope. RESULTS: In the capsulotomized animals, the cross sectional area of Leydig cells was gradually increased from 30 days onwards. There was no obvious change in the nucleus diameter of Leydig cells. However, The Leydig cell number was significantly increased from day 30 onwards. CONCLUSION: In rats, testicular capsulotomy may induce hyperplasia/hypertrophy of Leydig cells in the testis. PMID- 11907627 TI - Effect of Sarcostemma acidum stem extract on spermatogenesis in male albino rats. AB - AIM: To evaluate the possible antifertility activity of Sarcostemma acidum (Roxb) Voigt. stem extract in male rats. METHOD: Male rats were given 70% methanol extract of S. acidum stem orally at dose levels of 50 and 100 mg/kg/day for 60 days. Fertility was evaluated with mating test. Sperm motility and sperm density in cauda epididymides were also assessed. Biochemical and histological analyses were performed on blood samples and on the reproductive organs. RESULTS: S. acidum stem extract resulted in an arrest of spermatogenesis without any systemic side effect. Sperm motility as well as sperm density was reduced significantly. Treatment caused a 80% reduction in fertility at the 50 mg dose and complete suppression of fertility at the 100 mg dose. There was no significant change in RBC and WBC count, hemoglobin, haematocrit, sugar and urea in the whole blood and cholesterol, protein and phospholipid in the serum. The protein and glycogen content of the testes, fructose in the seminal vesicle and protein in epididymides were significantly decreased. Cholesterol in the testes was elevated. Treatment at both of the doses caused a marked reduction in the number of primary spermatocytes (preleptotene and pachytene), secondary spermatocytes and spermatids. The number of mature Leydig cells was decreased, and degenerating Leydig cells was increased proportionately. CONCLUSION: S. acidum stem extract arrests spermatogenesis in male rats without noticable side effects. PMID- 11907630 TI - Penile venous anatomy: application to surgery for erectile disturbance. AB - AIM: The structure of the human penile venous system has been well studied, but disappointing outcomes of penile venous surgery in certain patients have called into question on the anatomy. We planned to extend the anatomic knowledge with the ultimate goal of improving operative success. METHODS: Thirty-five patients, who had undergone penile venous surgery, complained of poor erection developed gradually 6 months to 7 years postoperatively. Cavernosography was performed again during their return visit. Seven new patients underwent spongiosography followed by immediate cavernosography. Eleven male cadavers were carefully dissected. The anatomical findings were applied to venous surgery in 155 patients, who were then followed with the International Index of Erectile Function Questionnaire-5 (IIEF-5). RESULTS: Imaging observation demonstrated that the deep dorsal vein served as a common vessel of the corpora cavernosa and corpus spongiosum. A prominent cavernosal vein was found coursing along each corpus cavernosum distally to the glans, in contrast to its reported description as a short segment at the penile hilum. All cadavers had two sets of para arterial veins sandwiching the dorsal artery. In 148 men available for follow-up, their mean IIEF-5 score was 9.3 preoperative and increased to 22.7 after the operation. The 88.5% (131/148) of the patients believed that venous stripping was a worthy treatment modality. Five cases required sildenafil to maintain their potentia, which was not working preoperatively. CONCLUSIONS: The failure of penile venous surgery has traditionally been ascribed to penile vein regeneration. However, our finding of a long and independent cavernosal vein and an independent set of para-arterial veins may be the principal cause in patients experiencing poor postoperative results. PMID- 11907629 TI - Expression of extracellular matrix proteins and vimentin in testes of azoospermic man: an immunohistochemical and morphometric study. AB - AIM: To investigate the changes in the extracellular matrix protein expression and the morphology of seminiferous tubules in the testis of 88 azoospermic men. METHODS: The patients were of the following categories: (1) 22 cases of Sertoli cell-only syndrome, (2) 20 cases of spermatogenic arrest, and (3) 46 cases with hypospermatogenesis. Testicular sections were immunohistochemically stained for fibronectin, vimentin, laminin and collagen type IV. The seminiferous tubular diameter and the connective matrix zone (CMZ, the acellular zone between the basement membrane [BM] and the peritubular cells) thickness were measured. Seminiferous tubules were typed according to the thickness of the connective matrix in the lamina propria. The predominant tubule type and the Johnsen and Silber scores were determined. RESULTS: The mean tubular diameter were 119 +/- 27, 117 +/- 20, and 140 +/- 38 microm for Groups 1, 2, and 3, respectively. Both the laminin and the type IV collagen were localized to the epithelial BM and peritubular cells. In most of the tubules, BM and peritubular cells were separated by a homogenous acellular layer, the CMZ, in which laminin, type IV collagen, fibronectin and vimentin were not present. It is perceived that the worse the testicular histology, the higher the thickness of the CMZ. CONCLUSION: In testis with no or low sperm production, the diameter of the seminiferous tubules is decreased, the thickness of the seminiferous tubular wall is increased and a CMZ is formed between the peritubular cells and the BM. The thickness of CMZ is increasing with the advancement of testiclar deterioration. The most important morphologic predictive factor for spermiogenesis is the predominant PMID- 11907631 TI - Risk of connective-tissue disease in men with testicular or penile prostheses: a preliminary study. AB - AIM: To help clarifying the possibility of connective-tissue diseases in men with penile or testicular prostheses. METHODS: Eight patients underwent inflatable penile prostheses and 15, testicular prostheses consented to the study. Their medical records were reviewed and a follow-up interview and physical and serological examinations were performed. RESULTS: In patients with penile prostheses, there was no abnormal antinuclear antibody (ANA) or IgM elevation. The serum levels of the rheumatoid factor (RF), C4, IgA and IgG were abnormal in one patient, and the levels of erythrocyte sedimentation rate (ESR) and C3, abnormal in two. Four had elevated IgE. In patients with testicular prostheses, there was no abnormal RF, ANA or IgM. The serum levels of ESR and IgA were abnormal in two, and three had abnormal C4, ten abnormal C3, and eleven decreased IgG. All had increased IgE. Men with penile prostheses had higher serum levels of IgG and IgM than those with testicular prostheses (P=0.001, P=0.016, respectively). The rates of abnormal values of IgE and IgG were higher in men with testicular prostheses than in men with penile prostheses (P=0.008, P=0.009, respectively). Physical examination was normal in all patients and nobody had documented symptoms pertinent to connective-tissue diseases. CONCLUSION: Our findings suggest that the risk of connective-tissue diseases is not higher in patients wearing prostheses as the ANA is negative and there is no apparent manifestation suggestive of connective-tissue diseases. PMID- 11907632 TI - Coenzyme Q10 levels in pigeon (Columba livia) spermatozoa. AB - AIM: To assess the CoQ10 levels in pigeon spermatozoa and to verify their possible correlation with spermatic kinetic parameters. METHODS: In pigeons the sperm motility percentage (MOT%), mean linear velocity (VCM) and morphology were determined in ejaculated semen. In addition intracellular CoQ10 concentrations were also detected with a HPLC method. RESULTS: Intracellular CoQ10 levels demonstrated wide individual variations, averaging 4.85 +/- 2.31 (SD) ng/106 spermatozoa. Statistical analysis showed a positive correlation of the substance with the sperm concentration (r= 0.63; P<0.05) and with the VCM (r=0.66; P<0.05), and a negative correlation with the MOT% (r = -0.78; P=0.01). No correlation was found between the CoQ10 concentration and the percentage of normal spermatozoa. CONCLUSION: Our results suggest a possible role for CoQ10 as a "fertility marker" in pigeons, which may be employed to monitor the pharmacological effects of cytostatic substances often used to reduce the pigeon fertility in urban environment. PMID- 11907633 TI - Effect of papaya seed extract on contractile response of cauda epididymal tubules. AB - AIM: To evaluate the administration of Carica papaya seed extract on the contractility of cauda epididymal tubules in male rats. METHODS: Adult male albino rats were administered intramuscularly papaya seed extract at a dose of 0.5 mg/kg/day for 7 days. Animals were killed, cauda epididymal tubules of 5 cm length were isolated and the contractile response to different concentrations of adrenalin (1-500 microg/25mL) was examined. In another group of animals, the contractile response was assayed 3 months after withdrawal of the treatment. RESULTS: Papaya seed extract brought about a significant decrease in the contractile response of epididymal tubules as compared with the control. After three months of papaya withdrawal, a nearly normal pattern of contraction was regained. CONCLUSION: Papaya seed treatment reversibly reduces the contractile response of cauda epididymal tubules. PMID- 11907634 TI - Morbidity and mortality in cardiovascular disorders: impact of reduced heart rate. PMID- 11907635 TI - Is it rational, reasonable or excessive, and consistently applied? One view of the increasing FDA emphasis on safety first for the release and use of antiarrhythmic drugs for supraventricular arrhythmias. PMID- 11907636 TI - Efficacy of amiodarone for the termination of chronic atrial fibrillation and maintenance of normal sinus rhythm: a prospective, multicenter, randomized, controlled, double blind trial. AB - OBJECTIVE: We sought to assess the efficacy and safety of amiodarone for restoration and maintenance of sinus rhythm in patients with chronic atrial fibrillation in a prospective, randomized, double blind trial. BACKGROUND: Restoration and preservation of sinus rhythm is difficult in patients with chronic atrial fibrillation. The efficacy of oral amiodarone has not been conclusively established. METHODS: Ninety-five patients with chronic atrial fibrillation, lasting an average of 35.6 months, were randomized to either amiodarone (600 mg/d) (47 patients) or placebo (48 patients) during four weeks. Nonresponders underwent electric cardioversion, and those who reverted continued with amiodarone (200 mg/d) or placebo. End-points were successful cardioversion and sinus rhythm maintenance. RESULTS: Sixteen patients (34.04%) in the amiodarone group reverted within 27.28 +/- 8.85 days in comparison with 0% in the placebo group (P < 0.000009). The conversion rate rose to 51.72% in patients with chronic atrial fibrillation lasting less than 12 months. Twenty-eight patients in the amiodarone group and 39 in the placebo group underwent electric cardioversion, which was successful in 19 patients (67.8%) of the amiodarone group and in 15 (38.46%) of the placebo group (P = 0.017). Altogether, conversion was obtained in 79.54% of the amiodarone group patients and in 38.46% of the placebo group patients (P < 0.0001). During follow-up, atrial fibrillation relapsed in 13 (37.14%) of 35 patients of the amiodarone group within 8.84 +/- 8.57 months and in 12 (80%; P = 0.009) of 15 patients of the placebo group within 2.74 +/- 3.41 months. CONCLUSIONS: Oral amiodarone restored sinus rhythm in one third of patients with chronic atrial fibrillation, increased the success rate of electric cardioversion, decreased the number of relapses and delayed their occurrence. PMID- 11907637 TI - Pharmacokinetics and pharmacodynamics of fluvastatin in heart transplant recipients taking cyclosporine A. AB - During the last decades, transplantation has become an established tool for the treatment of terminal organ failure. Beside immunological factors, hyperlipidemia is the main problem after heart transplantation, causing rapid transplant coronary artery disease (TxCAD) and poor long-term prognosis at the beginning of the transplantation. Heart transplant recipients are now effectively treated with lipid lowering substances, of which HMG-CoA-reductase inhibitors are the most potent. However, treatment with these substances correlates with an increased risk for the development of rhabdomyolysis due to therapy with the immunosuppressive cyclosporine A. Our study monitored the safety and efficacy of treatment with the HMG-CoA reductase inhibitor fluvastatin in heart transplant recipients compared to healthy controls. We investigated 10 patients receiving immunosuppressive therapy consisting of cyclosporine A, prednisone, and azathioprine who had increased concentrations of LDL-cholesterol (LDL-C), and 10 age-matched healthy controls. The patients were treated with 40 mg/day fluvastatin for 4 weeks and 20 mg/day for 4 additional weeks. Control individuals received 40 mg/day fluvastatin for 4 weeks only. Parameters of fluvastatin pharmacokinetics (maximum concentration of the drug (C(max.)), time (t(max.)) to reach C(max.), area under the concentration vs. time curve (AUC(0h-24h)), elimination half-life time (t(1/2))), apparent total body clearance (CL), blood cyclosporine A concentration, plasma lipids, and safety parameters were determined in both study groups at the beginning of the study and after 4 weeks. The latter were determined in the patient group also after 8 and 12 weeks. Treatment with 40 mg/day fluvastatin caused a significant decrease in total cholesterol (patients: 5.47 +/- 1.32 mmol/L vs. 7.30 +/- 1.83 mmol/L; controls: 4.69 +/- 0.64 mmol/L vs. 5.81 +/- 0.72 mmol/L), LDL-C (patients: 3.28 +/- 1.25 mmol/L vs. 5.00 +/- 1.85 mmol/L; controls: 2.58 +/- 0.63 mmol/L vs. 3.50 +/- 0.70 mmol/L), and triglycerides (patients: 1.99 +/- 0.77 mmol/L vs. 2.50 +/- 1.00 mmol/L; controls: 1.24 +/- 0.46 mmol/L vs. 1.72 +/- 0.67 mmol/L) in both study groups, whereas HDL-C was not significantly changed (patients: 1.29 +/- 0.35 mmol/L vs. 1.17 +/- 0.32 mmol/L; controls: 1.55 +/- 0.30 mmol/L vs. 1.53 +/- 0.26 mmol/L). Values of C(max.) and AUC(0h-24h) were higher in the patient group than in the control group (day 1, patients vs. controls, C(max.): 869.4 +/- 604.0 ng/mL vs. 211.9 +/- 113.9 ng/mL; AUC(0h-24h): 1948.8 +/- 1347.9 ng/mL*h vs. 549.4 +/- 247.4 ng/mL*h), whereas the corresponding value of CL was lower in the patient group (33.3 +/- 24.5 L/h vs. 107.9 +/- 95.8 L/h), and the values of t(max.) and t(1/2) showed no differences. In addition, values of C(max.) and AUC(0h-24h) after administration of 40 mg/day fluvastatin for 4 weeks in both groups were slightly higher than at the beginning, whereas the value of CL was slightly lower (day 28, patients vs. controls, C(max.): 1530.4 +/- 960.4 ng/mL vs. 254.7 +/- 199.8 ng/mL; AUC(0h-24h): 2615.3 +/- 1379.4 ng/mL*h vs. 841.8 +/- 421.4 ng/mL*h; CL: day 28, 21.4 +/- 15.3 L/h vs. 61.5 +/- 36.6 L/h). Except for an intermittent increase of creatine kinase, safety parameters showed no increases within the observation period. Our data suggest that fluvastatin effectively lowers plasma concentrations of cholesterol and LDL-C in patients after heart transplantation, however, the metabolism of fluvastatin is affected by concomitant therapy with cyclosporine A. Serum concentrations of fluvastatin should be monitored in cases of concomitant therapy with other substances interfering in the metabolism by competing cytochrome enzymes. PMID- 11907639 TI - Myocardial bradykinin following acute angiotensin-converting enzyme inhibition, AT1 receptor blockade, or combined inhibition in congestive heart failure. AB - BACKGROUND: The present study examined the effects of acute angiotensin converting enzyme inhibition (ACEI), AT(1) receptor blockade (AT(1) block), or combined treatment on in vitro and in vivo bradykinin (BK) levels. METHODS: BK levels were measured in isolated porcine myocyte preparations (n = 13) in the presence of exogenous BK (10(-8) M); with an ACEI (benezaprilat; 0.1 mM) and BK; an AT(1) block (valsartan; 10(-5) M) and BK; and combined treatment and BK. In a second study, myocardial microdialysis was used to measure porcine interstitial BK levels in both normal (n = 14) and pacing-induced congestive heart failure (CHF) (240 beats/min, 3 weeks, n = 16) under the following conditions: baseline, following ACEI (benezaprilat, 0.0625 mg/kg) or AT(1) block (valsartan, 0.1 mg/kg), and a combined treatment (benezaprilat, 0.0625 mg/kg; valsartan, 0.1 mg/kg). RESULTS: In the left ventricular myocyte study, BK levels increased over 93% with all treatments compared to untreated values (P < 0.05). In the in vivo study, basal interstitial BK values were lower in the CHF group than in controls (2.64 +/- 0.57 vs 5.91 +/- 1.4 nM, respectively, P < 0.05). Following acute infusion of the ACEI, BK levels in the CHF state increased from baseline (57% +/- 22; P < 0.05). Following combined ACEI/AT(1) block, BK levels increased from baseline in both control (42% +/- 11) and CHF groups (60% +/- 22; P < 0.05 for both). CONCLUSION: These findings suggest that ACEI, or combined ACEI/AT(1) block increased BK at the level of the myocyte and potentiated BK levels in the CHF myocardial interstitium. PMID- 11907638 TI - Role of desethylamiodarone in the anticoagulant effect of concurrent amiodarone and warfarin therapy. AB - BACKGROUND: The concurrent use of amiodarone and warfarin inhibits metabolism of S-warfarinby cytochrome P450 (CYP) 2C9, thereby increasing the anticoagulant effect of warfarin. Amiodarone primarily inhibits CYP1A2 and CYP3A4, and desethylamiodarone primarily inhibits CYP2C9. We investigate whether a relationship exists between the plasma concentration of desethylamiodarone and anticoagulation when amiodarone is administered to patients receiving warfarin therapy. METHODS AND RESULTS: The correlation between the plasma concentration of either amiodarone or desethylamiodarone, and prolongation of prothrombin time international normalized ratio/dose of warfarin (Delta INR/Dose) on day 7 of amiodarone administration was studied in 25 patients (22-74 years old) with structural heart disease and refractory arrhythmias receiving stable warfarin therapy. RESULTS: No correlation was found between the plasma concentration of amiodarone and Delta INR/Dose, but a correlation was found between the plasma concentration of desethylamiodarone and Delta INR/Dose. CONCLUSIONS: It was suggested that inhibition of CYP2C9 by desethylamiodarone, the active metabolite of amiodarone, plays an important role in the interaction of warfarin and amiodarone. PMID- 11907640 TI - Local delivery of a hydrophobic heparin reduces neointimal hyperplasia after balloon injury in rat carotid but not pig coronary arteries. AB - BACKGROUND: Intimal hyperplasia following percutaneous interventional vascular procedures is a major cause of restenosis. Although heparin inhibits intimal hyperplasia, it has not proven clinically useful in part due to an inadequate duration of intramural drug residence. This study was designed to evaluate the efficacy of local delivery of hydrophobic heparin (PTIR-RS-1), exhibiting increased intramural binding, on neointimal hyperplasia after angioplasty injury. METHODS AND RESULTS: PTIR-RS-1 was delivered locally into rat carotid arteries at three doses: 0.1 mM (440 IU), 0.3 mM (1320 IU), or 1.0 mM (4400 IU). Animals were killed at 14 days. In the pig, the doses tested were the low dose in the rat and a high dose 1 log higher. Animals were killed 28 days later. Morphometric analysis was performed to evaluate the intima: media ratio in rats and the normalized neointimal area in pigs. In rats a significant reduction in neointimal to medial area ratio from 0.73 +/- 0.15 for control vs 0.80 +/- 0.27 for sodium heparin (P = NS) and 0.15 +/- 0.07 for the 0.1 mM PTIR-RS-1 dose (P < 0.008). In pigs, PTIR-RS-1 the high dose reduced the normalized neointimal area by 16%, a difference that was not statistically significant. CONCLUSIONS: Increased hydrophobicity of heparin reduced neointimal area following balloon overstretch injury in the rat carotid but not the pig coronary artery model. This study attests to the importance of performing studies evaluating the pharmacologic effect of local delivery of a medication in at least two animal models of restenosis. PMID- 11907641 TI - Delta opioid receptors inhibit vagal bradycardia in the sinoatrial node. AB - BACKGROUND: Methionine-enkephalin-arginine-phenylalanine (MEAP) is an endogenous opiate derived from the C-terminal sequence of the larger precursor molecule proenkephalin. This heptapeptide is abundant in the myocardium and has significant vagolytic activity when infused systemically. MEAP interrupted vagal bradycardia when it was delivered directly into the sinoatrial node by local microdialysis. This study was conducted to determine the opioid receptor responsible for the vagolytic effect of MEAP. METHODS AND RESULTS: Microdialysis probes were placed in the sinoatrial node of mongrel dogs and perfused at 5 microL/min. Increasing doses of MEAP were included in the nodal perfusate and approximately two thirds of the vagal bradycardia was inhibited with a maximal effect at 0.3 nmoles/microL and a half-maximal response near 0.1 nmoles/microL. When deltorphin II (a delta opioid receptor agonist) was infused into the sinoatrial node, more than 95% of the vagal bradycardia was eliminated at 0.3 nmoles/microL with the half-maximal response near 0.1 nmoles/microL, indicating that deltorphin II was more efficacious than MEAP. The maximal deltorphin II and MEAP effects were both similarly reversed by the paired infusion of increasing doses of the delta opiate receptor antagonist, naltrindole. Selected mu (endomorphin, super DALDA) and kappa (dynorphin, U50488) receptor agonists and mu (CTAP) and kappa (norBNI) receptor antagonists were completely ineffective in this system. CONCLUSIONS: These data suggest that the vagolytic effect of MEAP involves the activation of delta opiate receptors within the sinoatrial node. PMID- 11907643 TI - The Lower Rhine Society for natural history and medicine in Bonn. PMID- 11907642 TI - Endothelin, nephropathy, and blood pressure. PMID- 11907644 TI - Regulation of gene expression in lymphocytes and antigen-presenting cells by measles virus: consequences for immunomodulation. AB - Acute measles, a well known disease usually contracted during early childhood, is still the major cause of vaccine-preventable infant deaths worldwide. There are about 40 million cases of acute measles per year, with more than one million cases of infant death as a consequence of measles. These are mainly due to opportunistic infections which develop on the basis of a generalized suppression of the cellular immunity in the course and after the acute disease. Lymphopenia, a general proliferative unresponsiveness of T cells ex vivo and cytokine imbalance, are considered as major hallmarks of measles virus (MV) induced immunosuppression. These findings are compatible with modulation of T cell responses by viral interference with professional antigen-presenting cells such as dendritic cells or direct effects on T cells by suppression of survival or proliferation signals. In vitro, MV interaction causes a variety of effects on dendritic cells, including maturation and loss of their allostimulatory functions. Whether there is an additional impact on the quality of T cell responses is unknown as yet. It is clear, however, that surface interaction of lymphocytes with the MV glycoprotein complex is necessary and sufficient to induce a state of proliferative unresponsiveness in T cells. This surface contact mediated signal essentially interferes with the propagation of the interleukin 2 receptor signal by blocking the activation of the protein kinase B, also called Akt kinase, both in vitro and after experimental infection. PMID- 11907645 TI - Ribavirin's antiviral mechanism of action: lethal mutagenesis? AB - Ribavirin, an antiviral drug discovered in 1972, is interesting and important for three reasons: (a) it exhibits antiviral activity against a broad range of RNA viruses; (b) it is currently used clinically to treat hepatitis C virus infections, respiratory syncytial virus infections, and Lassa fever virus infections; and (c) ribavirin's mechanism of action has remained unclear for many years. Here we recount the history of ribavirin and review recent reports regarding ribavirin's mechanism of action, including our studies demonstrating that ribavirin is an RNA virus mutagen and ribavirin's primary antiviral mechanism of action against a model RNA virus is via lethal mutagenesis of the RNA virus genomes. Implications for the development of improved versions of ribavirin and for the development of novel antiviral drugs are discussed. PMID- 11907646 TI - Association of a functional inducible nitric oxide synthase promoter variant with complications in type 2 diabetes. AB - Complications of diabetes have a genetic influence. Since increased inducible nitric oxide synthase (iNOS) gene ( NOS2A) expression can contribute to tissue damage, NOS2A is a worthy candidate for such a role. We therefore tested a 4-bp insertion/deletion (+/-) polymorphism 0.7 kb upstream of NOS2A for association with complications in type 2 diabetes patients, and also performed transient transfection experiments to examine the effect of this variant on promoter activity in kidney cells in culture. We investigated 379 Caucasian type 2 diabetes patients of British/European descent, 93 of whom had microalbuminuria, 26 overt nephropathy, 46 retinopathy, and 73 clinical neuropathy. Genotyping for the variant was carried out by PCR and automated Genescan analysis. Transient transfection studies involved the renal HEK 293 cell line and luciferase reporter gene constructs containing 1.1 kb of 5'-flanking DNA from '+' or '-' allele homozygotes. We found that the '+' allele frequency in patients without microalbuminuria was 12%, but was 23% in those with microalbuminuria ( P=0.0005), and was 26% in those with nephropathy ( P=0.0007), 22% in those with retinopathy ( P=0.037), and 23% in those with neuropathy ( P=0.045). The odds ratios for homozygote +/+ to have microalbuminuria or nephropathy were 2.4 (95% CI 1.4-4.2, P=0.0023) and 5.4 (95% CI 1.8-16, P=0.0009), respectively. Luciferase reporter gene constructs containing 1 kb of NOS2A promoter DNA for each allele were made and sequence analysis confirmed that the +/- variation was the only sequence difference present. Transient transfection of these into HEK 293 cells revealed 25 times higher reporter gene activity for the '+' allele compared with the '-' allele. Gel shift analysis with 30mer oligonucleotides corresponding to each allele showed specific binding to nuclear extracts, being greater for the '+' allele. Thus the '+' allele of the NOS2A promoter variant may confer higher iNOS expression, and could contribute to complications of type 2 diabetes, especially in the approximately 5% of patients homozygous for this variant. PMID- 11907648 TI - Altered modulation of soluble guanylate cyclase in lymphocytes from patients with liver disease. AB - Studies of animal models suggest that the activation of soluble guanylate cyclase by nitric oxide is altered in liver disease. We studied 77 patients with liver disease and 17 controls, to investigate whether the activation of soluble guanylate cyclase is altered in lymphocytes from patients with liver disease. The basal content of guanosine 3',5'-cyclic monophosphate (cGMP) in lymphocytes was decreased both in patients with liver cirrhosis (by 52%) and in patients with chronic hepatitis (by 62%). Activation of soluble guanylate cyclase by nitric oxide was higher in lymphocytes from patients with cirrhosis (3100+/-1000% of basal) or with hepatitis (5200+/-2500% of basal) than in lymphocytes from controls (1200+/-500% of basal). cGMP in plasma was increased in patients with liver disease. Successful (but not unsuccessful) treatment with interferon of patients with hepatitis due to virus C reversed all the above alterations. Altered modulation of soluble guanylate cyclase by nitric oxide in liver disease may play a role in the hemodynamic alterations found in these patients. PMID- 11907647 TI - Renal damage and salt-dependent hypertension in aged transgenic mice overexpressing endothelin-1. AB - The recent development of endothelin-1 (ET-1) antagonists and their potential use in the treatment of human disease raises questions as to the role of ET-1 in the pathophysiology of such cardiovascular ailments as hypertension, heart failure, renal failure and atherosclerosis. It is still unclear, for example, whether activation of an endogenous ET-1 system is itself the primary cause of any of these ailments. In that context, the phenotypic manifestations of chronic ET-1 overproduction may provide clues about the tissues and systems affected by ET-1. We therefore established two lines of transgenic mice overexpressing the ET-1 gene under the direction of its own promoter. These mice exhibited low body weight, diminished fur density and two- to fourfold increases in the ET-1 levels measured in plasma, heart, kidney and aorta. There were no apparent histological abnormalities in the visceral organs of young (8 weeks old) transgenic mice, nor was their blood pressure elevated. In aged (12 months old) transgenic mice, however, renal manifestations, including prominent interstitial fibrosis, renal cysts, glomerulosclerosis and narrowing of arterioles, were detected. These pathological changes were accompanied by decreased creatinine clearance, elevated urinary protein excretion and salt-dependent hypertension. It thus appears that mild, chronic overproduction of ET-1 does not primarily cause hypertension but triggers damaging changes in the kidney which lead to the susceptibility to salt induced hypertension. PMID- 11907649 TI - Mutations in the TMPRSS3 gene are a rare cause of childhood nonsyndromic deafness in Caucasian patients. AB - Two loci for nonsyndromic recessive deafness located on chromosome 21q22.3 have previously been reported, DFNB8 and DFNB10. Recently a gene which encodes a transmembrane serine protease, TMPRSS3 or ECHOS1, was found to be responsible for both the DFNB8 and DFNB10 phenotypes. To determine the contribution of TMPRSS3 mutations in the general congenital/childhood nonsyndromic deaf population we performed mutation analysis of the TMPRSS3 gene in 448 unrelated deaf patients from Spain, Italy, Greece, and Australia who did not have the common 35delG GJB2 mutation. From the 896 chromosomes studied we identified two novel pathogenic mutations accounting for four mutant alleles and at least 16 nonpathogenic sequence variants. The pathogenic mutations were a 1-bp deletion resulting in a frameshift and an amino acid substitution in the LDLRA domain of TMPRSS3. From this and another study we estimate the frequency of TMPRSS3 mutations in our sample as 0.45%, and approximately 0.38% in the general Caucasian childhood deaf population. However, TMPRSS3 is still an important contributor to genetic deafness in populations with large consanguineous families. PMID- 11907651 TI - Clinical tests of gastrointestinal permeability that rely on the urinary recovery of enterally administered probes can yield invalid results in critically ill patients. PMID- 11907652 TI - Are we doing a good job: PRISM, PIM and all that. PMID- 11907653 TI - Epidemiology of sepsis and infection in ICU patients from an international multicentre cohort study. AB - OBJECTIVES: To examine the incidence of infections and to describe them and their outcome in intensive care unit (ICU) patients. DESIGN AND SETTING: International prospective cohort study in which all patients admitted to the 28 participating units in eight countries between May 1997 and May 1998 were followed until hospital discharge. PATIENTS: A total of 14,364 patients were admitted to the ICUs, 6011 of whom stayed less than 24 h and 8353 more than 24 h. RESULTS: Overall 3034 infectious episodes were recorded at ICU admission (crude incidence: 21.1%). In ICU patients hospitalised longer than 24 h there were 1581 infectious episodes (crude incidence: 18.9%) including 713 (45%) in patients already infected at ICU admission. These rates varied between ICUs. Respiratory, digestive, urinary tracts, and primary bloodstream infections represented about 80% of all sites. Hospital-acquired and ICU-acquired infections were documented more frequently microbiologically than community-acquired infections (71% and 86%, respectively vs. 55%). About 28% of infections were associated with sepsis, 24% with severe sepsis and 30% with septic shock, and 18% were not classified. Crude hospital mortality rates ranged from 16.9% in non-infected patients to 53.6% in patients with hospital-acquired infections at the time of ICU admission and acquiring infection during the ICU stay. CONCLUSIONS: The crude incidence of ICU infections remains high, although the rate varies between ICUs and patient subsets, illustrating the added burden of nosocomial infections in the use of ICU resources. PMID- 11907655 TI - Pitfalls in gastrointestinal permeability measurement in ICU patients with multiple organ failure using differential sugar absorption. AB - OBJECTIVE: To assess whether gastrointestinal permeability (GIP) at intensive care unit (ICU) admission, measured by differential sugar absorption, is related to severity of disease and multiple organ failure (MOF). Post hoc, to analyse the relation between the urinary sugar recovery and renal function. DESIGN: Prospective observational cohort study. SETTING: Eighteen-bed general ICU of a teaching hospital. PATIENTS: Sixty-four ventilated patients admitted with MOF. INTERVENTIONS: GIP was assessed within 24 h using cellobiose (C), sucrose (S) and mannitol (M) absorption. MEASUREMENTS AND RESULTS: Severity of disease: APACHE II and III, SAPS II and MPM II systems. Organ failure: SOFA, MODS and Goris score. The median urinary recovery of C was 0.147% (range 0.004-2.145%), of S 0.249% (0.001-3.656%) and of M 10.7% (0.6-270%). In 16 patients, M recovery was over 100% of the oral dose. They received red blood cell transfusion (RBC). In the non transfused, the median cellobiose/mannitol (CM) ratio was 0.015 (0.0004-0.550). CM ratio was not related to severity of disease and inversely related to the SOFA score ( r=-0.30, p=0.04). Post hoc regression analysis showed that recoveries of C, S and M were positively related to urinary volume. Recoveries of C and S, but not of M, were positively related to creatinine clearance. The CM ratio corrected for diuresis, but was inversely related to creatinine clearance. CONCLUSIONS: Differential C, S and M absorption testing is unreliable after RBC transfusion, since bank blood contains mannitol. The excretion of C and S, but not of M, is limited by renal dysfunction. Differential sugar absorption is not reliable to test GIP in MOF patients, since non-permeability related factors act as confounders. PMID- 11907654 TI - Impact of sepsis, lung injury, and the role of lipid infusion on circulating prostacyclin and thromboxane A(2). AB - OBJECTIVE: To investigate whether plasma levels of prostacyclin (PGI2) and thromboxane A(2) (TxA2) are a function of the infusion rate of soybean-based fat emulsions, severity of systemic inflammation, and pulmonary organ failure. DESIGN: Prospective, randomized, crossover study. SETTING: Intensive care unit of a university hospital. PATIENTS: Eighteen critically ill patients, ten presenting with severe sepsis, eight with SIRS or sepsis complicated with ARDS. INTERVENTIONS: Patients were randomly assigned to receive rapid fat infusion over 6 h (rFI) or slow fat infusion over 24 h (sFI) along with parenteral nutrition. MEASUREMENTS AND RESULTS: The stable prostanoids 6-keto-PGF1alpha and TxB2 were measured in arterial and mixed venous blood samples, and at 6-h periods trans pulmonary balances (TPB) were calculated. Free linoleic acid fraction was determined in arterial blood. rFI induced greater increase of linoleic acid than sFI in both groups. Enhanced prostanoid levels and correlations with linoleic acid availabilities were found, however, in ARDS patients only, revealing the highest sepsis- and lung injury scores. Averaged TPB per 24 h was positive in the sepsis group and negative in the ARDS group as rFI induced lowest TPB values for TxB2 at 6 h. CONCLUSION: The quantity of prostanoids formed and their subsequent utilization are dependent on the availability of precursor linoleic acid and are probably affected by the severity of SIRS or sepsis and the existence of pulmonary organ failure, respectively. Because TxA2 might be extracted by the injured lung, rapid infusion of soybean-based fat emulsions should be avoided in patients suffering from severe pulmonary organ failure. PMID- 11907656 TI - Therapy of microcirculatory disorders in severe acute pancreatitis: what mediators should we block? AB - OBJECTIVE: To compare the effect of different vasoactive mediator antagonists in the same model of severe acute pancreatitis (AP) and to evaluate whether combinations of the agents exhibit synergistic effects. DESIGN: Prospective experimental study. SETTING: Microcirculation and pancreas research laboratory at an university hospital. PARTICIPANTS: Hundred eighty anesthetized male Sprague Dawley rats. INTERVENTIONS: Six hours after inducing AP by intra-ductal bile salt infusion and i.v. cerulein in 168 rats, these were randomized for therapy with (1) saline, (2) endothelin receptor antagonist (ET-RA), (3) platelet activating factor receptor antagonist (PAF-RA), (4) intercellular adhesion molecule-1 antibody (ICAM-1-AB) or different combinations (5-7). After 24 h the animals underwent a second laparotomy for intra-vital microscopic determination of pancreatic and colonic capillary permeability, blood flow and leukocyte endothelial interaction. RESULTS: AP induction decreased capillary blood flow and increased permeability and leukocyte rolling. ET-RA, PAF-RA and ICAM-1-AB decreased capillary permeability, increased blood flow and reduced leukocyte rolling. ET-RA was most effective in decreasing capillary permeability in both organs as well as in increasing pancreatic capillary blood flow. Combining vasoactive mediator blockers did not further improve target parameters. CONCLUSIONS: This study supports previous observations that ET-RA, PAF-RA and ICAM-1-AB improve microcirculation in AP and that ET-RA is more effective than PAF-RA or ICAM-1-AB, especially in counteracting capillary leakage. Although this may suggest that they act through different mechanisms, antagonist combinations failed to improve microcirculation further. We conclude that ET-RA is the most promising candidate for a clinical trial to reduce capillary leakage in patients with AP. PMID- 11907657 TI - The combination of a heat and moisture exchanger and a Booster: a clinical and bacteriological evaluation over 96 h. AB - OBJECTIVE: To determine whether the combination with a new device (Booster ) for active humidification improves the efficacy of a hydrophobic heat and moisture exchanger (HME). DESIGN AND SETTING: Prospective, interventional study in the ICU of a university hospital. PATIENTS: Consecutive patients requiring controlled mechanical ventilation INTERVENTIONS: Patients were ventilated with a HME, and a Booster was added for 96 h to the ventilatory circuit. MEASUREMENTS AND RESULTS: During the inspiration phase the following factors were measured: peak and mean airway pressures, maximal (beginning of inspiration), minimal (end of inspiration), and mean values of temperature of inspired gases, and relative and absolute humidity of inspired gases. Microbiological samples were obtained from the Booster, the ventilator side of the HME, and the tracheal secretions on days 1 and 4. Minimal and mean temperatures were increased as soon as the Booster was used and this increase was maintained for 96 h until the Booster was withdrawn. Then the temperature returned to baseline values. Absolute humidity values followed the same course. There was also some indirect evidence of very little, if any, changes in the HME resistance. The ventilatory side of the HMEs remained sterile in each patient, and the Booster was colonized by the same bacteria as those in the tracheal secretions. CONCLUSIONS: Adding the Booster to a hydrophobic HME improved the heat and water preservation of ventilatory gas. PMID- 11907658 TI - Mortality associated with late-onset pneumonia in the intensive care unit: results of a multi-center cohort study. AB - OBJECTIVE: To evaluate the attributable mortality associated with late-onset nosocomial pneumonia (LOP) while taking into account the severity at admission, the evolution of the patients during the first 4 days after admission to the ICU and the appropriateness of initial empiric antibiotic treatment. DESIGN: Multicenter cohort study with prospective standardization of diagnostic interventions when nosocomial pneumonia develops. SETTING: Medical and surgical ICUs of four university-affiliated teaching hospitals. PATIENTS: Seven hundred sixty-four consecutive patients requiring ICU hospitalization for at least 4 days. MAIN OUTCOME MEASURES: The clinical and biological data as well as the therapeutic data and the outcome were prospectively recorded from the day of admission to ICU discharge. Simplified Acute Physiologic Score (SAPS II) and Logistic Organ Dysfunction (LOD) score were collected and computed within the first 4 calendar days of ICU admission. Variables associated with the outcome were selected using a stepwise Cox model. The time to acquisition of the first LOP was then introduced in the final model as a time-dependent covariate. The analysis was stratified by ICU center. Finally, as initial antibiotic therapy could have an impact on the increased risk of death induced by LOP, the Cox model was applied again introducing LOP immediately adequately treated and LOP not immediately adequately treated as two different time-dependent covariates. RESULTS: Late-onset pneumonia developed in 89 patients (12%). A McCabe score of more than 1, SAPS II score and increases in SAPS between days 1 and 2, days 2 and 3, and days 3 and 4 were significantly associated with an increased risk of death. When the time to acquisition of the first episode of LOP was introduced into the Cox model, the LOP occurrence was associated with increased mortality, even adjusted over the selected prognostic parameters and after stratification by center (hazard ratio (HR)=1.53, 95% CI 1.02-2.3, p=0.04). When LOP immediately adequately treated and LOP not immediately adequately treated were separately introduced into the Cox model, inappropriately treated LOP remained significantly associated with an increased risk of mortality (HR=1.69, 95% CI 1.08-2.65, p=0.022), whereas appropriately treated LOP did not (HR=1.44, 95% CI 0.75-2.76, p=0.27). CONCLUSION: These data suggest that, in addition to severity scores, the underlying medical conditions and the evolution of severity within the first 4 days in ICU, late-onset pneumonia independently contribute to ICU patient mortality when empirical antibiotic treatment is not immediately appropriate. PMID- 11907659 TI - Physiological effects of constant versus decelerating inflation flow in patients with chronic obstructive pulmonary disease under controlled mechanical ventilation. AB - OBJECTIVE: To study the cardiorespiratory effects of inspiratory flow rate and waveform in COPD patients. DESIGN: Prospective physiological investigation with randomized allocations of experimental conditions. SETTING: A 14-bed medical ICU in a 1000-bed university hospital. PATIENTS AND PARTICIPANTS: Ten COPD intubated, sedated and paralyzed patients with chronic obstructive pulmonary disease (COPD), mechanically ventilated for acute respiratory failure. INTERVENTIONS: In volume controlled mode, three inflation flow rates of 0.40, 0.70, and 1.10 l/s for 20 min with a constant (CF) or a decelerating (DF) inflation flow profile. Each patient received all six experimental conditions in a random order. Tidal volume and respiratory frequency were similar during the experimental conditions. MEASUREMENTS AND RESULTS: Arterial blood gases, hemodynamics ( n=8), and respiratory mechanics were measured with zero end expiratory pressure. Between flow rates the median (25th-75th percentiles) values of PaO(2)/FIO(2) were 232 (132-289), 253 (161-338), 231 (163-352) for CF and 253 (143-331), 249 (164-360), 231 mmHg (187-351), for DF, respectively; the maximal airway pressures were 25.6, 28.3, 34.6 cmH(2)O for CF and 21.7, 29.6, 34.8 cmH(2)O for DF, respectively, the mean airway pressures were 8.9, 6.1, 5.4 cmH(2)O for CF and 9.1, 7, 6.5 cmH(2)O for DF, respectively. CONCLUSIONS: Changing the ventilator in volume-controlled mode with a DF or CF profile has no significant cardiorespiratory effect in intubated COPD patients mechanically ventilated for acute respiratory failure. PMID- 11907660 TI - Cognitive and perceived health status in patient with chronic obstructive pulmonary disease surviving acute on chronic respiratory failure: a controlled study. AB - OBJECTIVE: To evaluate the perceived health and cognitive status in survivors of chronic obstructive pulmonary disease (COPD) exacerbations requiring mechanical ventilation. DESIGN AND SETTING: Prospective controlled cohort study in a respiratory intermediate intensive care unit. PATIENTS: Sixty-three COPD patients at their first episode of acute on chronic respiratory failure requiring mechanical ventilation, controls were 34 consecutive stable COPD patients on long term oxygen therapy with no previous ICU admission. MEASUREMENTS AND RESULTS: Perceived health and cognitive status by means of the Nottingham Health Profile (NHP) and the Mini Mental State (MMS), respectively, at discharge and 3 and 6 months thereafter. At discharge patients showed significantly worse mean values than controls in MMS and in all NHP dimensions except pain. The MMS score was below 24, the threshold level of cognitive impairment, in 43% patients of the study group but only 3% of controls ( p=0.006). Six months after discharge patients showed NHP and MMS values similar to those of controls and the proportion of patients with a MMS score below 24 had significantly decreased to 17%, a value not significantly different from that in controls (5%). CONCLUSIONS: COPD patients surviving their first episode of acute on chronic respiratory failure requiring mechanical ventilation, suffer worsen perceived health status and cognitive function, which 6 months after discharge may improve to levels similar to those in stable COPD patients on long-term oxygen therapy with no previous ICU admission. PMID- 11907661 TI - Is the bispectral index appropriate for monitoring the sedation level of mechanically ventilated surgical ICU patients? AB - OBJECTIVE: To determine the value of the bispectral index (BIS) in assessing the depth of sedation in sedated and mechanically ventilated ICU patients, compared with clinical sedation scores. DESIGN AND SETTING: Prospective convenience sample in a 12-bed anesthesiological-surgical ICU of a university hospital. PATIENTS: 19 consecutive patients without any central neurological diseases requiring mechanical ventilation for more than 24 h. MEASUREMENTS: BIS version 3.12 and clinical depth of sedation assessed by the modified Observers's Assessment of Alertness/Sedation Scale, modified Glasgow Coma Scale, modified Ramsay Scale, Cook Scale, and Sedation-Agitation Scale were measured twice daily while patients were intubated and once daily after extubation until discharged from ICU. RESULTS: there was a moderate correlation between BIS and each sedation score in 11 patients (58%, "BIS patients") and no correlation in 8 patients (42%, "non-BIS patients"). We found no parameters distinguishing between these two groups. On average eight measurements were necessary to establish a statistical correlation. In the BIS patients the slopes of the linear regression curves showed significant differences for all BIS score combinations with increasing scattering at deeper sedation levels. CONCLUSIONS: BIS is correlated only in some ICU patients with the clinical assessment of their sedation level as based on various scores. At deeper sedation levels the interindividual differences increase. There were no criteria found to distinguish patients with and without correlation. This suggests that the BIS is not suitable for monitoring the sedation in a heterogeneous group of surgical ICU patients. PMID- 11907662 TI - Liver tissue oxygenation as measured by near-infrared spectroscopy in the critically ill child in correlation with central venous oxygen saturation. AB - OBJECTIVE: To evaluate the clinical usefulness of near-infrared spatially resolved spectroscopic quantitative assessment of liver tissue oxygenation for simple, non-invasive estimation of global tissue oxygenation in critically ill neonates and children. DESIGN: Prospective observational clinical study. SETTING: A tertiary multidisciplinary neonatal and paediatric intensive care unit (23 beds). PATIENTS: One hundred neonates and children consecutively admitted to the paediatric intensive care unit. MEASUREMENTS AND RESULTS: Near-infrared spectroscopic single-point assessment of liver tissue oxygenation index (TOI(Liver)) was compared with global tissue oxygenation as measured by central venous oxygen saturation (SvO(2)) and derived haemodynamic parameters. Data were compared using linear and multiple regression analysis. Overall correlation between TOI(Liver)and SvO(2) was good ( r=0.72, p<0.0001). Multivariable regression revealed that SvO(2) alone explained 51% of the observed variance of TOI(Liver). However, our data demonstrated large inter-individual differences between SvO(2) and TOI(Liver) values. CONCLUSION: Near-infrared spatially resolved spectroscopic quantitative measurement of liver tissue oxygenation correlates well with SvO(2) in critically ill neonates and children. Large inter individual SvO(2) to TOI(Liver) differences may prevent its use for non-invasive single-point estimation of global tissue oxygenation. Further clinical studies are required to validate the method with other regional and global haemodynamic parameters and to evaluate its clinical use for continuous non-invasive haemodynamic monitoring. PMID- 11907663 TI - Airway obstruction and ventilator dependency in young children with congenital cardiac defects: a role for self-expanding metal stents. AB - OBJECTIVE: Young children with congenital cardiac defect and airway obstruction leading to ventilator dependency present a significant clinical challenge with uncertain outcome. DESIGN: Retrospective review of our experience with self expanding metal stents in these young children between 1996-2000. RESULTS: Airway stenting has been undertaken in five such children (four boys, one girl) at our institution. Their mean age was 7.4 months (range 2-14 months), and four of the five had undergone congenital cardiac surgery and could not be weaned from the ventilator following surgery. These five children were ventilator dependent for a mean of 112 days (range 40-210 days, median 71). A total of ten self-expanding metal stents were inserted (4-11 mm in diameter and 15-33 mm in length). The sites stented included the trachea (two stents), the left main bronchus (three stents) and the bronchus intermedius (five stents). Four of these five children were successfully weaned from the ventilator and extubated after a mean time interval of 6 days (range 2-11 days, median 5.5) after stenting. One child failed to wean from the ventilator, required tracheostomy and is ventilator dependent after 8 months. There was one death 2 months after extubation but unrelated to the airway. Three children remain well and asymptomatic 24, 36 and 54 months after stenting. CONCLUSIONS: Airway stenting in such young children is an infrequent procedure often undertaken in dire circumstances. We have found it valuable in enabling ventilator-dependent children to be extubated with encouraging early results. Their long-term outlook remains uncertain and is dependent on the underlying cardiac status. PMID- 11907664 TI - Equal increases in respiratory system elastance reflect similar lung damage in experimental ventilator-induced lung injury. AB - OBJECTIVE: We hypothesized that a 50% increase in respiratory system elastance (Ers) would indicate similar degree of lung damage (equi-damage, ED), independently of ventilation strategy. DESIGN AND SETTING: A prospective, randomized animal laboratory investigation at a university hospital laboratory. SUBJECTS: 35 anesthetized, paralyzed, mechanically ventilated male Sprague-Dawley rats. INTERVENTIONS: Each rat was ventilated with a different combination of tidal volume, positive end-expiratory pressure, and inspired fraction of oxygen. Ers was determined throughout the experiment; the studies were interrupted when Ers reached 150% (ED) of its baseline value, or after 5 h. MEASUREMENTS AND RESULTS: Lung wet to dry weight ratio (W/D) was assessed. Morphological damage of the lung was scored on a grading of perivascular edema, hemorrhage, and breaks in the alveolar septa to obtain a total injury score. Twenty-four rats achieved an Ers of 150%: nine within 1 h (class 1), nine in 1-2 h (class 2), and six in 2-5 h (class 3). Eleven rats did not reach the target 50% increase in Ers (class 4). W/D was higher in rats that reached the target than in those that did not. W/D did not differ among rats that reached ED. Similarly, the total injury score did not differ among classes 1-3 but was higher than class 4. CONCLUSIONS: In the setting of VILI a 50% increase in Ers corresponds to an equal level of lung damage, irrespective of ventilatory setting and time of ventilation. PMID- 11907665 TI - Scoring systems in pediatric intensive care: PRISM III versus PIM. AB - OBJECTIVE: To compare the performance of two different clinical scoring systems that were developed to assess mortality probability in pediatric intensive care. DESIGN AND METHODS: Prospective cohort study in a multidisciplinary tertiary pediatric intensive care unit. The Pediatric Risk of Mortality score (PRISM III) and the Pediatric Index of Mortality (PIM) were collected for each patient. Standardized mortality rate (SMR), discrimination and calibration of both scoring systems were compared by goodness-of-fit tests and receiver operating characteristic (ROC) curves. RESULTS: Data from 303 patients were collected over a 9-month period. Twenty patients (6.6%) died in the PICU. Expected mortality based on PRISM III (12 h) was 6.96% (SMR 0.95; 95% CI 0.68-1.23), based on PRISM III (24 h) was 6.95% (SMR 0.95; 0.67-1.22) and based on PIM was 7.5% (SMR 0.88; 0.55-1.20). Calibration by Hosmer-Lemeshow goodness-of-fit test showed for PRISM III (12 h) chi(2) (8) =10.8, p=0.21; for PRISM III (24 h) chi(2) (8) =13.3, p=0.21 and for the PIM score chi(2) (8) = 4.92, p=0.77. Discriminatory performance assessed by ROC curves showed an area under the curve of 0.78 (95% CI 0.67-0.89) for the PRISM III score both after 12 and 24 h and 0.74 (0.63-0.85) for the PIM score. CONCLUSION: PRISM III and PIM scores are both adequate indicators of mortality probability for heterogeneous patient groups in pediatric intensive care. Possibly in larger studies (equivalence trial) a significant and relevant difference between these scores would be demonstrated. PMID- 11907666 TI - Serum procalcitonin monitoring for differential diagnosis of ventriculitis in adult intensive care patients. AB - The objective of our study was to assess the value of serum procalcitonin (PCT) monitoring in the differential diagnosis of ventriculitis in adult intensive care (ICU) patients. We analyzed 15 consecutive patients with ventriculitis in which a ventricular catheter had been inserted and contrasted these data with the observations in 10 patients with community-acquired bacterial meningitis. Cerebrospinal fluid (CSF) and blood samples were collected daily to assess serum PCT, C-reactive protein (CRP) and CSF leukocyte count. PCT levels were normal or slightly elevated in patients with ventriculitis with either positive or negative CSF bacterial culture but elevated in patients with bacterial meningitis. A PCT cut-off value of 1.0 ng/ml or more showed a specificity of 77% and a sensitivity of 68% for ventriculitis with positive CSF bacterial culture. Serum PCT levels reflected more accurately the time phases of disease during therapy. We conclude that the monitoring of serum PCT alone is not helpful for the differential diagnosis of ventriculitis, in contrast to that of bacterial meningitis. The value of PCT as an additional marker with which to assess the efficacy of therapy in ventriculitis is suggested, but requires further assessment. PMID- 11907667 TI - Bispectral index variations during tracheal suction in mechanically ventilated critically ill patients: effect of an alfentanil bolus. AB - OBJECTIVE: To evaluate the impact of an alfentanil dose on bispectral index (BIS) variations during tracheal suction in ICU sedated patients. DESIGN AND SETTING: A prospective open-label pilot study in a 12-bed surgical ICU in a university affiliated, tertiary referral hospital. PATIENTS: Eleven sedated (midazolam plus fentanyl) mechanically ventilated patients. INTERVENTIONS: Continuous monitoring of BIS with arterial pressure and heart rate before, during, and after tracheal suction without (control period) and with an intravenous bolus of alfentanil (15 microg/kg, alfentanil period) before suction. RESULTS: Steady-state BIS value was 61+/-8 for the control period and 59+/-7 for the alfentanil period. Blood pressure and heart rate were similar between baseline periods. One minute after tracheal suction, a significant increase in BIS level was observed in the control period, which remained significantly different from the alfentanil period until 10 min later. Significant higher systolic and diastolic blood pressure and heart rate were observed during the control period than the alfentanil period. However, no difference in Ramsay scores was observed between the two periods. CONCLUSIONS: An alfentanil bolus of 15 microg/kg markedly reduced the increase in BIS values, blood pressure, and heart rate observed immediately after tracheal suction. Therefore BIS monitoring in ICU may help to improve analgesia during invasive events. PMID- 11907668 TI - Cefepime-induced neurotoxicity: an underestimated complication of antibiotherapy in patients with acute renal failure. AB - OBJECTIVES: To describe five new cases of life-threatening cefepime-induced neurotoxicity observed in a 2-year period. SETTING: A university intensive care unit. PATIENTS: Five patients recently treated with cefepime, admitted for seizures and coma. All suffered from acute renal failure, induced by sepsis and combined aminoside therapy, or by cefepime itself in one case. INTERVENTIONS: All patients underwent hemodialysis, which led to complete neurological improvement in four of them. One patient remained comatose and subsequently died. MEASUREMENTS: Blood and CSF cefepime levels were measured by high performance liquid chromatography before and after hemodialysis. CONCLUSION: The frequency of cefepime-induced neurotoxicity is probably underestimated. Monitoring of renal function and close neurological survey in treated patients should allow an early diagnosis of this complication. Urgent hemodialysis seems the best therapeutic method to obtain a rapid neurological improvement. PMID- 11907669 TI - Cardiomyocyte apoptosis contributes to the pathology of the septic shock heart. PMID- 11907671 TI - Transtracheal open ventilation in respiratory care of patients after cardiovascular surgery. PMID- 11907672 TI - Closed system endotracheal suctioning maintains lung volume during volume controlled mechanical ventilation. PMID- 11907673 TI - Persistent severe hypoglycemia in acute zinc phosphide poisoning. PMID- 11907675 TI - Phototrophic consortia: model systems for symbiotic interrelations between prokaryotes. AB - Most symbiotic prokaryotes known to date have been found in association with eukaryotes, whereas only few (3.5%) bacteria or archaea are known that have established interactions with other prokaryotes. As revealed by direct microscopic investigations, however, multiple morphotypes of highly structured associations of different prokaryotes exist in nature. These so-called consortia appear to represent the most developed type of bacterial interaction. Phototrophic consortia are associations of green sulfur bacteria that surround a central chemotrophic bacterium. In some natural environments, almost all cells of green sulfur bacteria occur in the associated state, i.e. as epibionts of phototrophic consortia. In contrast to earlier speculations, the central bacterium belongs to the beta-Proteobacteria. Within the consortia, the green sulfur bacterial epibionts grow photolithoautotrophically, utilizing exogenous sulfide as photosynthetic electron donor. The entire consortium does not appear to be independent of organic carbon compounds, since it exhibits chemotaxis towards 2-oxoglutarate and, as demonstrated by microautoradiography, can also incorporate this compound. Intact consortia exhibit a scotophobic response in which the bacteriochlorophylls of the epibionts function as light sensors, whereas the chemotrophic central bacterium confers motility upon the association. Hence, a signal exchange must occur between the different bacteria. Based on their 16S rRNA gene sequences, the epibionts represent distinct phylotypes that are often only distantly related to known species of green sulfur bacteria. Since phototrophic consortia have recently become available in enrichment cultures, they can now serve as suitable model systems for the investigation of the molecular mechanisms of cell-cell recognition and signal exchange, and for studies of the coevolution of nonrelated prokaryotes. PMID- 11907676 TI - Signal peptides of secreted proteins of the archaeon Sulfolobus solfataricus: a genomic survey. AB - Analysis of the recently completed genome sequence of the thermoacidophilic archaeon Sulfolobus solfataricus reveals that about 4.2% of its proteome consists of putative secretory proteins with signal peptides. This includes members of the four major classes of signal peptides: secretory signal peptides, twin-arginine signal peptides, possible lipoprotein precursors, and type IV pilin signal peptides. The latter group is surprisingly large compared to the size of the groups in other organisms and seems to be used predominately for a subset of extracellular substrate-binding proteins. PMID- 11907677 TI - Correlation of long-range membrane order with temperature-dependent growth characteristics of parent and a cold-sensitive, branched-chain-fatty-acid deficient mutant of Listeria monocytogenes. AB - Listeria monocytogenes is a food-borne, pathogenic, psychrotolerant bacterium that grows at refrigeration temperatures. Long-range membrane order of the parent (10403S) and of a cold-sensitive mutant ( cld-1) deficient in odd-numbered, branched-chain fatty acids was measured using the width of the central line of spectra of an electron paramagnetic resonance probe, 4,4-dimethyl-2-heptyl-2 hexyloxazolidine- N-oxyl (7N14), that locates deep in the hydrocarbon region of the membranes. The line width decreased from 0.9 to 0.5 milliTesla (mT) over the temperature range of 0-10 degrees for strain 10403S and -5 to 32 degrees C for strain cld-1 independent of protein state (heat denatured or intact). This provided new evidence for phase transitions in the membranes. When strain cld-1 was grown in medium supplemented with 2-methylbutyric acid, which restores anteiso fatty acids and the ability to grow at low temperature, the change in central line width as a function of temperature resembled that of strain 10403S. The temperatures at which the central line width became 0.8 mT corresponded to those at which growth became very slow in both strains (3-5 degrees C for 10403S, 15 degrees C for cld-1) as determined by Arrhenius plots. These data underscore the critical role of odd-numbered anteiso fatty acids in influencing the lower temperature limits of growth through their effects on long-range membrane fluidity. PMID- 11907678 TI - Membrane association of Klebsiella pneumoniae NifL is affected by molecular oxygen and combined nitrogen. AB - In the diazotroph Klebsiella pneumoniae, NifL and NifA regulate transcription of the nitrogen fixation genes in response to molecular oxygen and combined nitrogen. We recently showed that Fnr is the primary oxygen sensor. Fnr transduces the oxygen signal towards the negative regulator NifL by activating genes whose products reduce the FAD moiety of NifL under anoxic conditions. These Fnr-dependent gene products could be membrane-bound components of the anaerobic electron transport chain. Consequently, in this study we examined the localization of NifL within the cell under various growth conditions. In K. pneumoniae grown under oxygen- and nitrogen-limited conditions, approximately 55% of the total NifL protein was found in the membrane fraction. However, when the cells were grown aerobically or shifted to nitrogen sufficiency, less than 10% of total NifL was membrane-associated. In contrast to NifL, NifA was located in the cytoplasm under all growth conditions tested. Further studies using K. pneumoniae mutant strains showed that, under derepressing conditions but in the absence of either the primary oxygen sensor Fnr or the primary nitrogen sensor GlnK and the ammonium transporter AmtB, NifL was located in the cytoplasm and inhibited NifA activity. These findings suggest that under nitrogen- and oxygen-limitation, a significantly higher membrane affinity of NifL might create a spatial gap between NifL and its cytoplasmic target protein NifA, thereby impairing inhibition of NifA by NifL. Localization of GlnK further showed that, under nitrogen-limited conditions but independent of the presence of oxygen, 15-20% of the total GlnK is membrane-associated. PMID- 11907680 TI - Isolation, characterization, and fermentative pattern of a novel thermotolerant Prototheca zopfii var. hydrocarbonea strain producing ethanol and CO2 from glucose at 40 degrees C. AB - A novel thermotolerant strain of the achlorophyllous micro-alga Prototheca was isolated from a hot spring. The isolate was found to produce an appreciable amount of ethanol and CO2 from glucose under anoxic conditions at both 25 and 40 degrees C; this type of alcohol fermentation has not yet been reported in the genus Prototheca. Moreover, it also evolved gas from sucrose after a time lag at 40 degrees C. Its taxonomic characteristics coincided with those of Prototheca zopfii var. hydrocarbonea, and phylogenetic analysis, based on a small-subunit (SSU) rDNA sequence, also revealed a close relationship between the two strains. D-lactic acid, ethanol, CO2 and a trace of acetic acid were produced from glucose, but L-lactic acid, formic acid, and H2 were not. At 25 degrees C, D lactic acid and ethanol were produced in approximately equimolar amounts under N2/H2/CO2, whereas ethanol production was predominant under N2. More ethanol was produced at 40 degrees C than at 25 degrees C irrespective of the gas composition in the atmosphere. This is the first report on gas production from glucose and on the changes in the fermentative patterns as a function of temperature for the genus Prototheca. PMID- 11907679 TI - Anaerobic degradation of n-hexane in a denitrifying bacterium: further degradation of the initial intermediate (1-methylpentyl)succinate via C-skeleton rearrangement. AB - The anaerobic degradation pathway of the saturated hydrocarbon n-hexane in a denitrifying strain (HxN1) was examined by gas chromatography-mass spectrometry of derivatized extracts from cultures grown with unlabeled and deuterated substrate; several authentic standard compounds were included for comparison. The study was focused on possible reaction steps that follow the initial formation of (1-methylpentyl)succinate from n-hexane and fumarate. 4-Methyloctanoic, 4 methyloct-2-enoic, 2-methylhexanoic, 2-methylhex-2-enoic and 3-hydroxy-2 methylhexanoic acids (in addition to a few other methyl-branched acids) were detected in n-hexane-grown but not in n-hexanoate-grown cultures. Labeling indicated preservation of the original carbon chain of n-hexane in these acids. Tracing of the deuterium label of 3- d1-(1-methylpentyl)succinate in tentative subsequent products indicated a deuterium/carboxyl carbon exchange in the succinate moiety. This suggests that the metabolism of (1-methylpentyl)succinate employs reactions analogous to those in the established conversion of succinyl CoA via methylmalonyl-CoA to propionyl-CoA. Accordingly, a pathway is proposed in which (1-methylpentyl)succinate is converted to the CoA-thioester, rearranged to (2-methylhexyl)malonyl-CoA and decarboxylated (perhaps by a transcarboxylase) to 4-methyloctanoyl-CoA. The other identified fatty acids match with a further degradation of 4-methyloctanoyl-CoA via rounds of conventional beta-oxidation. Such a pathway would also allow regeneration of fumarate (for n-hexane activation) from propionyl-CoA formed as intermediate and hence present a cyclic process. PMID- 11907681 TI - Inositol is specifically involved in the sexual program of the fission yeast Schizosaccharomyces pombe. AB - The fission yeast Schizosaccharomyces pombe is a natural auxotroph for inositol and fails to grow in the complete absence of it. It was previously reported that a small concentration of inositol in the culture medium supports vegetative growth, but not mating and sporulation, and a tenfold of that concentration also supports mating and sporulation. The purpose of the present work was to investigate whether a moderate inositol starvation specifically affected events of the sexual program of development. A homothallic culture grown to the stationary phase in medium with a small inositol concentration was sterile but cells in the stationary phase of growth synchronously entered and completed the sexual cycle when inositol was added, without need of previous cell divisions. This suggests the involvement of inositol in a mechanism (or mechanisms) of the sexual program. The events of the program that were affected by inositol starvation were investigated. Commitment to mating and production of pheromone M were shown not to be inositol-dependent. A diploid strain homozygous at the mating-type locus and carrying a pat1-114 temperature-sensitive mutation in homozygous configuration sporulated under inositol starvation at the restrictive temperature; therefore starvation did not directly affect meiosis or sporulation. In contrast, production of pheromone P and the response of cells to pheromones were found to be inositol-dependent. The possibility that inositol or one of its derivative compounds is involved in pheromone P secretion and in pheromone signal reception is discussed. PMID- 11907682 TI - Morphological and physiological changes in Streptomyces lividans induced by different yeasts. AB - Streptomyces development is a complex process that eventually finishes with the formation of individual unigenomic spores from the aerial hyphae. Intraspecific and interspecific signals must play a key role in triggering or blocking this process. Here we show that interaction between two types of microorganisms, Streptomyces and yeasts, leads to alteration of the Streptomyces developmental program. This alteration is due to the action of invertase produced by the yeast on the sucrose present in the culture media, making glucose and fructose readily available for growth. PMID- 11907683 TI - Characterization of a novel insertion sequence, IS Bp1, in Burkholderia pseudomallei. AB - During screening for antigenic proteins in Burkholderia pseudomallei, a novel insertion sequence, IS Bp1, was found by sequence similarity searches. IS Bp1 contains two overlapping ORFs of 261 bp ( orfA) and 852 bp ( orfB), encoding 87 and 284 amino acid residues, respectively, and an imperfect inverted repeat. The putative protein encoded by orfA (OrfA) is similar to the OrfA in insertion sequences of the IS 3 family in other bacteria, showing 49% and 76% amino acid identity and similarity, respectively, with the transposase encoded by IS D1 of Desulfovibrio vulgaris vulgaris. The putative protein encoded by orfB (OrfB) is similar to the OrfB in insertion sequences of the IS 3 family in other bacteria, showing 43% and 62% amino acid identity and similarity, respectively, with the transposase encoded by IS 1222 of Enterobacter agglomerans. Sequence analysis of OrfA showed the presence of an alpha-helix-turn-alpha-helix motif, as well as the putative leucine zipper at its 3' end, for possible DNA binding to the terminal inverted repeats. Sequence analysis of OrfB showed the presence of a DDE motif of aspartic acid, aspartic acid, and glutamic acid, a highly conserved motif present in OrfB of other members of the IS 3 family. Furthermore, several other conserved amino acid residues, including the arginine residue located seven amino acids downstream from the glutamic acid residue, were observed. PCR amplification of the IS Bp1 gene showed a specific band in 65% of the 26 B. pseudomallei strains tested. Southern blot hybridization after XhoI or SacI digestion showed nine different patterns of hybridization. The number of copies of IS Bp1 in those strains that possessed the insertion sequence ranged from three to 12. Using several insertion sequences and a combination of insertion-sequence-based and non insertion-sequence-based methods such as ribotyping will probably increase the discriminatory power of molecular typing in B. pseudomallei. PMID- 11907685 TI - Perceptual segregation of overlapping shapes activates posterior extrastriate visual cortex in man. AB - Objects in natural scenes are rarely seen in isolation, but are usually overlapping or partially occluding other objects. To recognize individual objects, the visual system must be able to segregate overlapping objects from one another. Evidence from lesions in humans and monkeys suggest that perceptual segregation of occluded or overlapping objects involves extrastriate visual cortex. In monkeys, area V4 has been shown to play an important role in recognizing occluded or poorly salient shapes. In humans, a retinotopic homologue of ventral V4 (V4v) has been described, but it is not known whether this area is also functionally homologous to area V4 in monkeys. In this study, we tried to localize the visual cortical regions involved in perceptual segregation of overlapping shapes using positron emission tomography (PET). Regional cerebral blood flow (rCBF) was measured in seven subjects while they discriminated the relative areas of simultaneously presented rectangular shapes. In the control condition, the shapes were displayed without overlaps; in a second condition, the shapes overlapped each other partially. In a third condition, the shapes did not overlap but had been reduced in salience by adding random noise to the stimuli. Contrasting the overlapping shape condition with the control condition identified a single region in the left posterior lateral occipital cortex. The rCBF in this region also increased, though more weakly, during discrimination of shapes embedded in noise, relative to the control condition. The region activated by segregation of overlapping shapes was located in the posterior occipital cortex close to the anterior border of area V2, near the average location of human V4v as determined by retinotopic mapping studies. The activation of this region of extrastriate visual cortex by a task that involved segregation of overlapping shapes is consistent with monkey V4 and human V4v being functionally homologous. We conclude that discrimination of overlapping shapes involves in particular a region of extrastriate visual cortex located in the left lateral occipital cortex and that this region may correspond to human V4v. PMID- 11907684 TI - A novel Streptomyces gene, samR, with different effects on differentiation of Streptomyces ansochromogenes and Streptomyces coelicolor. AB - A 1.4-kb DNA fragment from Streptomyces ansochromogenes accelerated mycelium formation of S. ansochromogenes when present on a multicopy plasmid. The DNA fragment contains one complete open reading frame, designated samR, encoding a protein with 213 amino acids that contains a likely DNA-binding helix-turn-helix motif close to its N-terminus. The deduced SamR protein resembles the product of the hppR gene, which is involved in the regulation of catabolism of 3-(3 hydroxyphenyl) propionate in Rhodococcus globerulus. A samR disruption mutant was constructed that presented a bald phenotype and failed to form aerial hyphae and spores. We suggest that samR plays an important role in the emergence of aerial hyphae from substrate mycelium. An almost identical gene of Streptomyces coelicolor was also subjected to gene disruption. Surprisingly, the mutant was able to develop an aerial mycelium, but it remained white and deficient in sporulation instead of forming gray spores. PMID- 11907686 TI - Structure of joint variability in bimanual pointing tasks. AB - Changes in the structure of motor variability during practicing a bimanual pointing task were investigated using the framework of the uncontrolled manifold (UCM) hypothesis. The subjects performed fast and accurate planar movements with both arms, one moving the pointer and the other moving the target. The UCM hypothesis predicts that joint kinematic variability will be structured to selectively stabilize important task variables. This prediction was tested with respect to selective stabilization of the trajectory of the endpoint of each arm (unimanual control hypotheses) and with respect to selective stabilization of the timecourse of the vectorial distance between the target and the pointer tip (bimanual control hypothesis). Components of joint position variance not affecting and affecting a mean value of a selected variable were computed at each 10% of normalized movement time. The ratio of these two components ( R(V)) served as a quantitative index of selective stabilization. Both unimanual control hypotheses and the bimanual control hypothesis were supported both prior to and after practice. However, the R(V) values for the bimanual control hypothesis were significantly higher than for either of the unimanual control hypothesis, suggesting that the bimanual synergy was not simply a simultaneous execution of two unimanual synergies. After practice, an improvement in both movement speed and accuracy was accompanied by counterintuitive changes in the structure of kinematic variability. Components of joint position variance affecting and not affecting a mean value of a selected variable decreased, but there was a significantly larger drop in the latter when applied on each of the three selected task variables corresponding to the three control hypotheses. We conclude that the UCM hypothesis allows quantitative assessment of the degree of stabilization of selected performance variables and provides information on changes in the structure of a multijoint synergy that may not be reflected in its overall performance. PMID- 11907687 TI - The effects of human ankle muscle vibration on posture and balance during adaptive locomotion. AB - This study investigated the contribution of ankle muscle proprioception to the control of dynamic stability and lower limb kinematics during adaptive locomotion, by using mechanical vibration to alter the muscle spindle output of individuals' stance limbs. It was hypothesised that muscle length information from the ankle of the stance limb provides information describing location as well as acceleration of the centre of mass (COM) with respect to the support foot during the swing phase of locomotion. Our prediction, based on this hypothesis was that ankle muscle vibration would cause changes to the position and acceleration of the COM and/or compensatory postural responses. Vibrators were attached to both the stance limb ankle plantarflexors (at the Achilles tendon) and the opposing dorsiflexor muscle group (over tibialis anterior). Participants were required to walk along a 9-m travel path and step over any obstacles placed in their way. There were three task conditions: (1) an obstacle (15 cm in height) was positioned at the midpoint of the walkway prior to the start of the trial, (2) the same obstacle was triggered to appear unexpectedly one step in front of the participant at the walkway midpoint and (3) the subjects' walking path remained clear. The participants' starting position was manipulated so that the first step over the obstacle (when present) was always performed with their right leg. For each obstacle condition participants experienced the following vibration conditions: no vibration, vibration of the left leg calf muscles or vibration of the anterior compartment muscles of the lower left leg. Vibration began one step before the obstacle at left leg heel contact and continued for 1 s. Vibrating the ankle muscles of the stance limb during the step over an obstacle resulted in significant changes to COM behaviour [measured as displacement, acceleration and position with respect to the centre of pressure (COP)] in both the medial/lateral (M/L) and anterior/posterior planes. There were also significant task-specific changes in stepping behaviour associated with COM control (measured as peak M/L acceleration, M/L foot displacement and COP position under the stance foot during the step over the obstacle). The results provide strong evidence that the primary endings of ankle muscle spindles play a significant role in the control of posture and balance during the swing phase of locomotion by providing information describing the movement of the body's COM with respect to the support foot. Our results also provide supporting evidence for the proposal that there are context dependent changes in muscle spindle sensitivity during human locomotion. PMID- 11907688 TI - Old age impairs the use of arbitrary visual cues for predictive control of fingertip forces during grasp. AB - Old age impairs the ability to form new associations for declarative memory, but the ability to acquire and retain procedural memories remains relatively intact. Thus, it is unclear whether old age affects the ability to learn the visuomotor associations needed to set efficient fingertip forces for handling familiar objects. We studied the ability for human subjects to use visual cues (color) about the mechanical properties (texture or weight) of a grasped object to control fingertip forces during prehension. Old and young adults (mean age 77 years and 22 years, respectively) grasped and lifted an object that varied in texture at the gripped surfaces (experiment 1: sandpaper versus acetate surface materials) or weight (experiment 2: 200 g versus 400 g). The object was color coded according to the mechanical property in the "visual cue" condition, and the mechanical property varied unpredictably across lifts in the "no visual cue" condition. In experiment 1 (texture), the young adults' grip force (force normal to the gripped surface) when the object lifted from the support surface was 24% smaller when the surfaces were color-coded. The old adults' grip force did not vary between the visual conditions despite their accurate reports of the grip surface colors prior to each lift. Comparable findings were obtained in experiment 2, when object weight was varied and peak grip force rate prior to object lift-off was measured. Furthermore old and young subjects alike used about 2 N of grip force when lifting the 200 g object in experiment 2. Therefore, the old adults' failure to adjust grip force when the color cue was present cannot be attributed to a general inability or unwillingness to use low grip force when handling objects. We conclude that old age affects the associative learning that links visual identification of an object with the fingertip forces for efficiently handling the object. In contrast, old and young subjects' grip force was influenced by the preceding lift, regardless of visual cues, which supports existing theories that multiple internal representations govern predictive control of fingertip forces during prehension. PMID- 11907689 TI - Eyeblink conditioning indicates cerebellar abnormality in dyslexia. AB - There is increasing evidence that cerebellar deficit may be a causal factor in dyslexia. The cerebellum is considered to be the major structure involved in classical conditioning of the eyeblink response. In a direct test of cerebellar function in learning, 13 dyslexic participants (mean age 19.5 years) and 13 control participants matched for age and IQ undertook an eyeblink conditioning experiment in which for 60 acquisition trials an 800-ms auditory tone (conditioned stimulus, CS) was presented. On 70% of the trials an 80-ms corneal airpuff (unconditioned stimulus, US) was presented 720 ms after the tone onset. The cerebellar deficit hypothesis, uniquely of the causal hypotheses for dyslexia, predicts that the dyslexic participants would show abnormal performance in the incidence and/or timing of the conditioned response (CR) of an eyeblink in response to the tone (and before the US). Three of the dyslexic group showed no conditioning at all. Furthermore, the dyslexic group showed significantly worse "tuning" of the CR timing, together with significantly reduced habituation of the orienting response (OR) to the CS. Individual analyses indicated that 85% of the dyslexic group showed either no conditioning or abnormally poor CR tuning and/or abnormally low OR habituation. It is concluded that the findings provide further converging evidence of cerebellar abnormality in dyslexia and for the first time demonstrate that there are fundamental abnormalities in the way that dyslexic people learn. Equally important, the eye blink conditioning paradigm provides a method of investigating further both homogeneity and heterogeneity in the learning deficits underlying dyslexia and other developmental disorders. PMID- 11907690 TI - Pattern of cortical reorganization in amyotrophic lateral sclerosis: a functional magnetic resonance imaging study. AB - Depending on individual lesion location and extent, reorganization of the human motor system has been observed with a high interindividual variability. In addition, variability of forces exerted, of motor effort, and of movement strategies complicates the interpretation of functional imaging studies. We hypothesize that a general pattern of reorganization can be identified if a homogeneous patient population is chosen and experimental conditions are controlled. Patients with amyotrophic lateral sclerosis (ALS) and healthy volunteers were trained to perform a simple finger flexion task with 10% of each individual's maximum grip force with constant movement amplitude and frequency. The activation pattern in ALS patients was distinctly different to that in healthy controls: In ALS patients, motor cortex activation was located more anteriorly, encompassing the premotor gyrus. The cluster volume within the supplementary motor area (SMA) was higher and shifted toward the pre-SMA. Contralateral inferior area 6 and bilateral parietal area 40 revealed higher cluster volumes. Our results demonstrate a general pattern of functional changes after motor neuron degeneration. They support the concept of a structurally parallel and functionally specialized organization of voluntary motor control. Degeneration of the first and second motor neurons leads to enhanced recruitment of motor areas usually involved in initiation and planning of movement. Partial compensation between functionally related motor areas seems to be a strategy to optimize performance if the most efficient pathway is unavailable. PMID- 11907691 TI - Factors influencing variability in load forces in a tripod grasp. AB - The aim of this investigation was to determine the factors that influence the variability in the load force during a precision grip. Subjects grasped an object with three coarse contact surfaces using a tripod grasp of the thumb, index and middle fingers. The orientations of the contact surfaces and their locations relative to the object's center of mass were varied. Forces and torques exerted by each of the three digits were measured or computed from the equilibrium conditions and analyzed during the static hold phase. The static load force depends on the object's weight, but also on the precise location of the center of pressure of each digit, as well as on the magnitude of the tangential torque and horizontal grip force. Variations in these parameters caused deviations in the load forces by as much as 20% or more from their nominal values. For the thumb, the largest source of variability was the vertical location of the center of pressure. For the two fingers, variations in the locations of the centers of pressure (along the horizontal and vertical) and in the magnitudes of the tangential torques had effects that were about equal in magnitude. Since many of the sources of variability in load force are unpredictable, these results point to the importance of online feedback regulation of grip force. PMID- 11907692 TI - A re-examination of the possibility of controlling the firing rate gain of neurons by balancing excitatory and inhibitory conductances. AB - It has been suggested that balancing excitatory and inhibitory conductance levels can control the firing rate gain of single neurons, defined as the slope of the relation between discharge frequency and excitatory conductance. According to this view the increase in firing rate produced by an input pathway can be controlled independently of the ongoing firing rate by adjusting the mixture of excitatory and inhibitory conductances produced by other pathways converging onto the neuron. These conclusions were derived from a simple RC-neuron model with no active conductances, or firing threshold mechanism. The analysis of that model considered only the subthreshold behaviour and did not consider the relation between total trans-membrane conductance and firing rate. Similar conclusions were also derived from a simple parallel conductance based model. In this paper I consider, as an example of a repetitively firing neuron, a generic model of cat lumbar alpha-motoneurons with excitatory and inhibitory inputs and a second independent excitatory pathway. The excitatory and inhibitory inputs can be thought of as central descending controls while the second excitatory pathway may represent, for example, the monosynaptic Ia-afferent pathway. I have re-examined the possibility that the firing rate gain of the "afferent" pathway can be controlled independently of the ongoing firing rate by balancing the excitatory and inhibitory conductances activated by the descending inputs. The steady state firing rate of the model motoneuron increased nearly linearly with the excitatory current, as it does in real motoneurons (primary firing range). The model motoneuron also showed a secondary firing range, whose slope was steeper than in primary range. The firing rate gain was measured by increasing the conductance of the "afferent" pathway. The firing rate gain (in the primary and secondary firing range) of the "afferent" pathway was found to be the same regardless of the particular mixture of excitatory and inhibitory conductances acting to produce the ongoing firing rate. This result was obtained for a single-compartment model, as well as for a two-compartment model consisting of an active somatic compartment and a dendritic compartment containing an L-type calcium conductance. Put simply, the firing rate gain of an input to a neuron cannot be controlled by balancing excitatory and inhibitory conductances produced by other independent input pathways, or by the spatial distribution of excitation and inhibition across the neuron. Three potential ways of controlling the firing rate gain are presented in the "Discussion". Firing rate gain can be controlled by actions at the presynaptic terminal, by inhibitory feedback, which is a function of the neuron's firing rate, or by neuromodulator substances that affect intrinsic inward or outward currents. PMID- 11907693 TI - Areas of the human brain activated by ambient visual motion, indicating three kinds of self-movement. AB - In a positron emission tomography (PET) study, a very large visual display was used to simulate continuous observer roll, yaw, and linear movement in depth. A global analysis based on all three experiments identified brain areas that responded to the three conditions' shared characteristic of coherent, wide-field motion versus incoherent motion. Several areas were identified, in the posterior inferior temporal cortex (Brodmann area 37), paralimbic cortex, pulvinar, and midbrain tegmentum. In addition, occipital region KO was sensitive to roll and expansion but not yaw (i.e., coherent displays containing differential flow). Continuous ambient motion did not activate V5/MT selectively. The network of sites responding specifically to coherent motion contrasted with the extensive, contiguous activation that both coherent and incoherent motion elicited in visual areas V1, V2, and V3. The coherent motion mechanisms, furthermore, extended beyond the traditional dorsal pathway proposed to account for visual motion processing, and included subcortical and limbic structures, which are implicated in polysensory processing, posture regulation, and arousal. PMID- 11907694 TI - A method to evaluate reflex excitability of the human ankle plantarflexors despite changes in maximal activation capacities. AB - Stretch reflexes were evoked in the submaximally activated ankle extensors during sinusoidal length perturbations. A mean stretch reflex (SR) amplitude ((-)SRA), i.e., SR area/SR duration was quantified for the soleus muscle and for the gastrocnemii muscles. (-)SRA was also expressed in relative values ((-)SRA(rel)), i.e., SRA was related to the corresponding background electromyogram (EMG). Sinusoidal length perturbations were applied in two ways: (1) at a fixed frequency (16 Hz) and at different levels of voluntary contraction [10% to 70% maximal voluntary contraction (MVC)] and (2) at a constant activation level (50% MVC) and at frequencies ranging from 6 to 16 Hz. Then, new parameters are proposed to characterize muscle reflex excitability. The first parameter, SR(index), consisting in the slope of the (-)SRA-EMG relationship, was considered to be more representative of central influences on the reflex pathway. Secondly, the frequency distribution of the stretch reflex was analyzed and the area under the (-)SRA(rel)-frequency curve gave the second parameter (FD-(-)SRA(rel)). This parameter should account for both central and peripheral mechanisms on the reflex pathway. These parameters were found to be higher for the highly excitable soleus than for the less excitable gastrocnemii muscles. SR(index) and FD-(-)SRA(rel) can be proposed as a tool to analyze changes in reflex excitability, which can accompany a process of neuromuscular plasticity. In order to validate the procedure, the proposed parameters were quantified in a case study before and after a period of plyometric training. PMID- 11907695 TI - Vestibular contributions across the execution of a voluntary forward step. AB - This work addressed the influence of information arising from the vestibular system on the dynamic control of a forward step. Six subjects performed the stepping task with their eyes closed under three conditions of bipolar, binaural galvanic vestibular stimulation (GVS), including (1) no GVS, (2) GVS with the anode electrode on the side of the swing limb, and (3) GVS with the anode electrode on the side of the stance limb. GVS was delivered 1,500 ms prior to a cue to step. Ground reaction forces were collected from three force platforms and movement was recorded from IRED markers placed bilaterally on the body. The results showed that, following slight deviations caused by GVS onset, the step initiation behaviour was unaffected, but lateral deviations were found during the latter, more dynamic, phases of stepping for centre of mass trajectories, time integrals of the centre of pressure displacement and upper body roll. These findings showed that vestibular information is used differently across the execution of a step without vision. While the initiation phase is run in a feedforward manner without vestibular influence, vestibular information appears to be upregulated during the more dynamic phases. Also, the level of up regulation may be different across step execution. PMID- 11907696 TI - Interconnections of auditory areas in the guinea pig neocortex. AB - By studying the efferent projections of five auditory areas in the guinea pig cortex, we sought evidence that the larger fields can be divided into subareas based on unique patterns of cortical connections. Small extracellular injections of biocytin were made in combination with evoked potential mapping or single-unit analysis and histochemical determination of cortical landmarks. The two core fields, primary (AI) and dorsocaudal (DC), are partially surrounded by six adjacent belt areas, leaving two gaps: one at the rostral edge of AI and the other at the dorsal edge. All of the areas studied projected to their nearest neighbors, but AI was the only area to project to all seven of the other auditory areas. The caudal, high-frequency (more than 4 kHz) end of AI had different projections from the rostral, low-frequency (less than 1.5 kHz) end, and there was no evidence of connections between the two ends. Each end had separate dorsal and ventral projections. The two ends of AI may be working independently. By contrast, area DC had strong connections between its high- and low-frequency ends and it may be involved in auditory/visual integration. The dorsorostral belt (DRB) was subdivided into two zones on the basis of its projections: the more rostral part appears to overlap the second somatosensory area and be bimodal, while the caudal part has stronger auditory connections. The small belt area (area S) had separate physiological and anatomical properties from the rest of the rostral belt. PMID- 11907697 TI - Age effects in dynamic vision based on orientation identification. AB - Dynamic vision considers the time domain of visual processing, presumably a function of the magnocellular system. The temporal visual component is not completely developed at the beginning of school age. The effects of age on a certain aspect of dynamic visual perception are reported in this study of 286 subjects in the age range of 7-68 years. Three simple orientation identification tasks were used: a fixation, a saccade, and an anti task. In each case the subjects had to identify the last of a series of fast changing orientations of a T-symbol before it disappeared after a random time period. The percentage of correct identifications in 50-100 trials was measured for each of the three tasks. In the age range of about 14 to about 28 years the subjects could perform the tasks at 90% correct or better. At younger and at older ages the mean values of the scores are considerably lower due to a higher percentage of subjects with difficulties in the performance of the tasks. It is discussed whether it is the magnocellular system which mediates the dynamic orientation identification in these tasks and which begins to decline relatively early in life in parallel with a relative loss of saccade control. PMID- 11907698 TI - The validity of the inter- and/or intrahemispheric deficit hypothesis as an explanation of the co-occurrence of motor and language impairments. AB - From an initial cohort of 15 children, between the ages of 5 and 10 years, who had expressive oral language problems, 4 were shown in an earlier study to be both motor and language impaired. Two explanatory hypotheses were proposed to account for this communality: (a) cerebellar deficit; (b) inter- and/or intrahemispheric deficit. In order to explore the validity of the latter explanation, the same group of children, together with a matched control group, were required to carry out two sensory matching tests designed to tap inter- and intrahemispheric information processing abilities: hand-hand and foot-hand. The results, discussed in the light of Liederman's shielding model, provided more support for the hypothesis of an interhemispheric information processing problem from left to right rather than an intrahemispheric problem. PMID- 11907699 TI - Sigma smooth pursuit eye tracking: constant k values revisited. AB - Effective sigma tracking, i.e., apparent movement perception when slow eye movements are made across a stationary repetitive pattern under stroboscopic illumination, has been shown to be a function of the distance between sequential stimuli (P(s)) and the flash frequency (f(s)). The relationship between these factors and eye velocity ( V (e)) has been formally specified as V (e)= k P(s)f(s)[deg s(-1)], where it has been argued that the value of k, which defines the rate limit for eye velocity, is normally 1, or exceptionally 2 or 3. However, theoretically the limitations on the maximum value for k are the maximum optimal pursuit speed for eye tracking (V(max)) and the minimum values which P(s) and f(s) can assume while preserving target discrimination, and since the values for V(max) are known to lie well beyond 20 deg/s and those for P(s) and f s) well below 0.3 deg and 10 Hz respectively, it should be possible to demonstrate empirically that k can assume integer values considerably larger than the indicated maximum of 3. To test this prediction, three subjects performed seven series of five EOG-monitored trials producing sigma-pursuit, with values of k ranging from 1 to 7. All subjects evidenced smooth pursuit eye tracking for every condition and reported experiencing sigma-type apparent motion in 95% of the trials. The results confirm theoretical expectations and unequivocally demonstrate that sigma tracking can be readily effected under conditions where k significantly exceeds the maximal values previously reported, in conformity with theory. PMID- 11907700 TI - Control of human locomotion under various task constraints. AB - The goal of this study was to identify the control mechanism used for locomotion pointing regulation under different external temporal constraints. Subjects ( n=8) had to walk on a treadmill through a number of virtual hallways and cross a pair of gliding doors that opened and closed at a constant preset frequency (0.5 Hz or 1 Hz). Crossing performance, step durations, and step lengths were used as dependent measures. The results revealed the regulation of locomotion occurred earlier and was more pronounced at 0.5 Hz than at 1 Hz, making performance better at 0.5 Hz. Nevertheless at the two frequencies the control mechanism appears similar; it is grounded on information movement coupling. This control mechanism allows for the production of specific behavior according to the task constraints. PMID- 11907701 TI - Forearm bone mineral density in postmenopausal women with rheumatoid arthritis. AB - Osteoporosis in cases of rheumatoid arthritis (RA) is multifactorial, and the pathogenesis of bone loss induced by RA in postmenopausal women is not fully understood. The purpose of the present study was to determine the factors that affect forearm bone mineral density (BMD) in postmenopausal women with RA. In total, 839 postmenopausal women aged 46-90 years, were enrolled in the study; 470 patients with RA and 369 healthy controls (CON). Forearm (distal radius) BMD, measured by DXA using a DTX-200 (Osteometer, MediTech, CA, USA), was significantly lower in the RA group than in the CON group (P < 0.0001), even when adjusted for age, height, body weight, body mass index, and years since menopause (YSM) (P < 0.01). On multiple regression analysis, in the CON group, age and YSM were significantly correlated with BMD (P < 0.01 and P < 0.05, respectively). On the other hand, in the RA group, in addition to YSM, anatomic grade in the wrist, modified health assessment questionnaire (HAQ) score for the upper extremities, and erythrocyte sedimentation rate were each significantly correlated with BMD (P < 0.0001, P < 0.001, P < 0.0001, and P < 0.001, respectively), whereas functional class, duration of disease, dose of prednisolone used, modified HAQ score for the lower extremities, and the levels of c-reactive protein and rheumatoid factor were not. The present study with a large number of subjects shows that in addition to YSM, disuse (anatomic grade) of the wrist, arm function, and disease activity appear to be significant determinants of forearm BMD in postmenopausal women with RA. PMID- 11907702 TI - Low body size and elevated sex-hormone binding globulin distinguish men with idiopathic vertebral fracture. AB - Factors predisposing to vertebral fracture in men are less well defined compared with women. Most studies of osteoporosis in men have included patients with low bone mineral density (BMD), with or without vertebral fracture, or have included other fractures. To clarify these associations we investigated sex hormone levels, bone markers, and (indirectly) lean body mass (LBM) in 81 men with idiopathic vertebral fracture. Serum testosterone, estradiol, sex-hormone binding globulin (SHBG), 24-hr urinary creatinine (24-hr UCr), urinary free deoxypyridinoline (UfDPD) and serum type I procollagen carboxy-terminal propeptide, type I procollagen amino-terminal propeptide, type I collagen carboxy terminal telopeptide, and osteocalcin were measured. SHBG was higher and 24-hr UCr lower in osteoporotic subjects. UfDPD was higher when corrected for 24-hr UCr. Serum bone turnover markers were not significantly increased, nor were serum sex hormones (and free hormone indices) significantly decreased in patients. SHBG levels were inversely related with lumbar spine and femoral neck BMD in both patients and control subjects. Free estradiol index was only correlated with BMD in men with osteoporosis. Body size is lower in men with established osteoporosis. The normal free hormone indices suggest that SHBG does not affect free hormone levels whereas the relationship between SHBG (but not sex hormones) and 24-hr UCr points to a relationship between SHBG and LBM. The association of high levels of SHBG with low levels of LBM may indicate an action via the known inverse relationship of SHBG with IGF-I, though any action through IGF-I probably occurred at an earlier age than that at which the patients presented. Estrogen has no relationship with BMD in normal men but may play a role in men with osteoporosis. PMID- 11907703 TI - Oral contraceptives moderately effect bone resorption markers and serum-soluble interleukin-6 receptor concentrations. AB - This study investigated the effect of ethinylestradiol(EE2)-containing oral contraceptives on mineral and bone metabolism and on serum soluble-interleukin-6 receptor (sIL-6R) during the menstrual cycle. Twelve women, aged 24.3 +/- 2.9 years, were examined. Blood and 24-hour and fasting urine samples were obtained during one menstrual cycle between cycle day 3-5 (t(1)), cycle day 10-12 (t(2)), cycle day 24-26 (t(3)), and again on day 3-5 of the next cycle (t(4)). EE2 intake was 0 mg at t(1), 30 mg at t(2), 30 mg at t(3) and 0 mg at t(4). Fasting renal phosphorus and calcium excretions were slightly reduced at t(2) and t(3) compared with t(1) and t(4) (P < 0.05-0.001). Moreover, renal excretion of the bone resorption marker C-Teleopeptide was at t(3) reduced by 26% compared with t(1)(P < 0.01) and by 13% compared with t(4)(P > 0.05). Fasting sIL-6R levels were 16.5% lower at t(2) and 12% lower at t(3) than at t(4) (P < 0.01 and P < 0.05). sIL-6R was correlated with total deoxypyridinoline excretion (r = +0.35; P < 0.05) and with fasting renal excretions of calcium (r = +0.36; P < 0.05) and phosphorus (r = +0.29; P < 0.05). In summary, our data suggest that in young women, cyclic monthly oral contraceptive intake is associated with small, but significant variations in bone resorption processes and in serum sIL-6R levels. Results are a further indication that monthly fluctuations of bone resorption in young women are mediated by sex hormones and that osteoclastic activity is stimulated by cytokines in vivo. PMID- 11907705 TI - A potent 1,4-dihydropyridine L-type calcium channel blocker, benidipine, promotes osteoblast differentiation. AB - During their differentiation, osteoblasts sequentially express type I collagen, alkaline phosphatase (ALP), and osteocalcin, and then undergo mineral deposition. Among dihydropyridine-type calcium channel blockers, only benidipine stimulated ALP activity of osteoblastic cells derived from neonatal mouse calvaria. To identify the molecular target of benidipine and elucidate the mechanism of action of the drug in osteoblasts, the mouse osteoblastic cell line MC3T3-E1 was used. Benidipine prompted ALP activity and ALP transcription induced by ascorbic acid, and mineral deposition by ascorbic acid and b-glycerophosphate. Benidipine, however, did not change collagen accumulation. MC3T3-E1 cells expressed the L type Ca channel a1C subunit throughout the differentiation process, and Ca influx by potassium ions and Bay K 8644, an agonist, was strongly attenuated by benidipine. Each one of three structurally different classes of Ca channel blockers, nifedipine, verapamil, and diltiazem stimulated ALP activity, although at much higher concentrations of ca. 100 nM than benidipine, 1 pM. These results suggest that benidipine directly exerts its effect on osteoblasts and promotes osteoblast differentiation after the step of collagen accumulation by blocking the L-type Ca channel. Since benidipine blocked Ca influx more potently than the three other Ca channel blockers, the unique and potent osteoblast differentiating ability of benidipine may be due to its high affinity for Ca channel together with its high membrane retaining ability, as has been previously reported. PMID- 11907706 TI - Alendronate disturbs vesicular trafficking in osteoclasts. AB - The nitrogen-containing bisphosphonate alendronate inhibits osteoclast-mediated bone resorption through inhibition of the mevalonate pathway. This results in impaired protein prenylation and may affect the function of small GTPases in osteoclasts. Since these proteins are important regulators of vesicle transport in cells, we investigated the possible interference of alendronate with these processes in isolated rat osteoclasts. We show here that alendronate-induced inhibition of bone resorption coincides with accumulation of tartrate-resistant acid phosphatase- and electron dense material-containing tubular vesicles in osteoclasts. Alendronate-induced changes in osteoclasts also included widening of the sealing zone areas and incomplete organization of tight attachments and ruffled borders. Osteoclasts also appeared partially detached from the bone surface, and organic matrix was typically dissolved only at the edges of the resorption pits on alendronate-coated bone slices. In contrast, resorption pits on the control and clodronate-coated bone slices were thoroughly resorbed. Inhibition of bone resorption by alendronate was not, however, related to a decrease in osteoclast number. In conclusion, our findings suggest that alendronate inactivates osteoclasts by mechanisms that impair their intracellular vesicle transport, apoptosis being only a secondary phenomenon to this. PMID- 11907704 TI - Bone mineral density in femoral neck is positively correlated to circulating insulin-like growth factor (IGF)-I and IGF-binding protein (IGFBP)-3 in Swedish men. AB - Studies on the hormonal regulation of bone metabolism in men have indicated covariation between insulin-like growth factor-I (IGF-I) and sex hormones with bone mineral density (BMD). In this study the relationships between BMD in total body, lumbar spine, femoral neck, distal and ultradistal (UD) radius and circulating levels of IGFs, IGF binding proteins (IGFBPs), and sex steroids were investigated in 55 Swedish men between 22 and 85 (52 +/- 18, mean +/- SD) years of age. BMD in total body, distal and UD radius, and femoral neck was positively correlated with serum IGF-I (r = 0.31 to 0.49), IGF-II (r = 0.32 to 0.48), IGFBP 3 (r = 0.37 to 0.53), and free androgen index (FAI) (r = 0.32 to 0.40), and negatively with IGFBP-1 (r = -0.37 to -0.41) and IGFBP-2 (r = -0.29 to -0.41) levels. A positive correlation was observed between BMD in femoral neck and estradiol/SHBG ratio (r = 0.34, P = 0.01). Age correlated negatively with serum IGF-I, IGF-II, IGFBP-3, FAI, estradiol/SHBG ratio, and BMD in total body, distal and UD radius, and femoral neck, and positively with IGFBP-1, IGFBP-2, and SHBG levels. According to stepwise multiple regression analyses, a combination of weight, IGFBP-3, and testosterone accounted for 43% of the variation in BMD in femoral neck, 34% in ultradistal radius and 48% in total body (P < 0.0001). These findings indicate that sex hormones and the different components of the IGF system are associated with BMD in Swedish men, suggesting that age-related changes in these systems could contribute to the development of osteoporosis in elderly men. PMID- 11907707 TI - Olpadronate prevents the bone loss induced by cyclosporine in the rat. AB - The aim of the present in vivo experimental study was to investigate changes in bone turnover and bone mineral density (BMD) induced by cyclosporine (CsA) administration. The effectiveness of olpadronate (OPD) in preventing bone loss associated with CsA treatment was also evaluated. Forty male Sprague-Dawley rats (approximately 5 months old) were treated as follows: Group I: CsA+OPD vehicles (control); Group II: CsA 15 mg/kg + OPD vehicle; Group III: CsA 15 mg/kg + 4 ug OPD/100g rat; Group IV: CsA 15 mg/kg + 8 ug OPD/100g rat; Group V: CsA 15 mg/kg + 16 ug OPD/100g rat. CsA was administered by daily oral gavage and OPD by intraperitoneal injection once a week. Serum bone-alkaline phosphatase (b-ALP) and urinary deoxypyridinoline (DPyr) were measured on days 0, 14 and 30. Total skeleton, femur, lumbar spine, proximal, and middle tibia BMDs were measured on days 0 and 30. No significant differences were found between the CsA and the control groups as regards serum bALP levels, on days 14 and 30. CsA+OPD treated rats presented a transient increment in serum b-ALP on day 14 and a significantly lower level on day 30 compared to the control and CsA groups (P < 0.05). On days 14 and 30, DPyr excretion increased in the CsA group compared to control animals (P < 0.05). The three studied doses of OPD induced a significant decrease in DPyr excretion in the CsA group on days 14 and 30 (P < 0.05). Group V (receiving the highest dose of OPD) presented a significantly lower level of DPyr compared to the other two OPD-treated groups (P < 0.05). On day 30, the CsA group presented a significant reduction in proximal tibia, spine and whole femur BMDs (P < 0.05) compared to controls. On day 30, OPD treatment increased BMD of all the studied areas in CsA rats. Proximal tibia BMD of group V reached significantly higher values than the other studied OPD groups (P < 0.05). In summary, this study suggests that CsA-induced high bone resorption and trabecular bone loss is prevented by cotreatment with OPD. Moreover, it encourages the possible use of OPD to treat patients receiving CsA as immunosuppressive therapy. PMID- 11907709 TI - Predictors of bone mineral density in aging healthy men varies by skeletal site. AB - Factors contributing to the pathogenesis of osteoporosis in women are well defined. However, changes in bone mineral metabolism in aging men and the role of various factors in the pathogenesis of age-related bone loss in men are less well understood. To further clarify these changes, serum and urine biochemical parameters, and lumbar spine, hip, and total body bone mineral density (BMD) were evaluated in a small sample of 45 healthy men aged 20-80 years, and multiple regression models were developed to predict age-related bone loss. Serum calcium, phosphate, albumin, creatinine clearance, osteocalcin, C-terminal propeptide of type I procollagen, log-free androgen index, dehydroepiandrosterone sulfate (DHEA S), and androstenedione decreased with age, and serum sex hormone binding globulin and urine total and free pyridinoline increased with age. Femoral neck BMD decreased with age, but remained stable at the other sites measured. Multiple regression analysis indicated that serum phosphate, DHEA-S, and intact parathyroid hormone (PTH) predicted lumbar spine BMD. Age, serum phosphate, and PTH predicted femoral neck BMD. Urine-free deoxypyridinoline alone predicted femoral greater trochanter BMD. Weight, serum creatinine, and urine-free deoxypyridinoline predicted total body BMD. We conclude that predictor variables of bone density vary by skeletal site in healthy men. Alterations in adrenal androgens, phosphate, and PTH may be important in the pathogenesis of bone loss with aging in men. PMID- 11907708 TI - The first stage of transforming growth factor beta1 activation is release of the large latent complex from the extracellular matrix of growth plate chondrocytes by matrix vesicle stromelysin-1 (MMP-3). AB - Transforming growth factor beta-1 (TGF-beta1) is secreted in a biologically inactive form and stored in the extracellular matrix as a 290 kDa complex consisting of the mature TGF-beta1 homodimer (Mr 25 kDa), the latency-associated peptide (LAP; Mr 75 kDa), and the latent TGF-beta1 binding protein-1 (LTBP1; Mr 190 kDa). Latent TGF-beta1, composed of these three components, is known as the "large latent TGF-beta1 complex." In contrast, latent TGF-beta1 without LTBP1 is known as "small latent TGF-beta1." For all latent forms, dissociation of the TGF beta1 homodimer from LAP is necessary for growth factor activation and acquisition of biological activity. Matrix vesicles produced by growth plate chondrocytes contain matrix metalloproteinases that can activate small latent TGF beta1. The enzyme responsible for this is matrix metalloproteinase-3 (MMP-3), although matrix vesicles also contain MMP-2 and plasminogen activator. The present study tested the hypothesis that matrix vesicle enzymes are also involved in the release of the large latent TGF-beta1 complex stored in the extracellular matrix. Matrix vesicles were isolated from cultures of resting zone and growth zone chondrocytes and metalloproteinases present in the matrix vesicles extracted with guanidine-HCl. Chondrocyte extracellular matrices were prepared by lysing confluent cultures and removing the lysed cells. The matrices were incubated with matrix vesicle extracts and the release of total and active TGF-beta1 was determined. To determine if MMP-2 or MMP-3 was involved in the release, matrix vesicle extracts were preincubated with anti-MMP-2 antibody or anti-MMP-3 antibody to selectively deplete the enzyme activity. Matrices were also treated with rhMMP-2 or rhMMP-3. To determine the identity of the released protein(s), digests were separated on SDS-polyacrylamide gels and Western blotting analysis was performed using a specific antibody to LTBP1. Matrix vesicle extracts released both active and total (=latent + active) TGF-beta1 in a time-dependent manner, with peak release after 1 hour of incubation. The amount of total TGF beta1 released was 10 times higher than the release of active TGF-beta1. The effect of the matrix vesicle extracts was dose-dependent; in addition, the amount and ratio of active to total TGF-b1 released was very similar, irrespective of the source of matrix or matrix vesicle extracts. Pre-incubation of matrix vesicle extracts with anti-MMP-3 antibody blocked the release of active and total TGF beta1, whereas pre-incubation with pre-immune IgG or anti-MMP-2 antibody had no effect. The addition of rhMMP-3, but not rhMMP-2, caused a dose-dependent increase in the release of total, but not active, TGF-beta1. Western analysis confirmed that both matrix vesicle extracts and rhMMP-3 released the large latent TGF-beta1 complex from the matrix. In addition to the expected 290, 230, and 190 kDa bands, samples run without reduction also contained proteins of molecular weights 110 and 50 kDa that reacted with the anti-LTBP1 antibody. When these same samples were electrophoresed after reduction, the high molecular weight immunoreactive bands disappeared and three bands of molecular weight 75, 32, and 25 kDa were observed. These results indicate that matrix vesicles contain enzymes, especially MMP-3, which are responsible for the release of TGF-beta1 from the matrix, most of which is in latent form. Further, the data suggest that release of the large complex occurs via cleavage at several novel sites in the 130 kDa LTBP1 molecule. Since matrix vesicle MMP-3 is also able to activate small latent TGF-beta1, these results suggest that the large latent TGF-beta1 complex protects against activation of the small latent TGF-beta1. Thus, the data suggest that release of the large latent TGF-bl complex from the matrix and activation of the latent growth factor are only two steps of what must be at least a three-step process. PMID- 11907710 TI - Growth hormone secretion and bone mineral density in prepubertal black and white boys. AB - Racial differences in bone mineral density (BMD) appear to account in part for racial differences in the incidence of osteoporosis and fractures. We previously reported that the greater BMD in adult blacks compared with whites is associated with a higher serum 17 beta-estradiol and greater secretion of growth hormone (GH) in men but not women. To determine whether these racial differences occur in prepubertal boys, we measured spontaneous overnight GH secretion, serum testosterone, 17 beta-estradiol, IGF-I, and IGFBP3, IGF-I/ IGFBP3 ratio, BMD of the total body, forearm, lumbar spine, trochanter, and femoral neck, and lean body mass and body fat in 14 healthy black and 16 white boys ages 6-7 years. Measurements of GH were obtained at 20-minute intervals for 12 hours. Results were analyzed by deconvolution and are expressed as mean +/- SE. Whereas BMD of the hip (0.755 +/- 0.020 vs 0.663 +/- 0.021 g/cm(2), P = 0.0037), trochanter (0.617 +/- 0.014 vs 0.552 +/- 0.018 g/cm(2), P = 0.0102) and femoral neck (0.710+/-0.018 vs 0.6381 +/- 0.021 g/cm(2), P = 0.0157) were significantly greater in black compared with white boys, BMD of the total body (0.768 +/- 0.010 vs 0.741 +/- 0.012 g/cm(2), NS), forearm (0.405 +/- 0.010 vs 0.380 +/- 0.008 g/cm(2), NS), and lumbar spine (0.612 +/- 0.013 vs 0.609 +/- 0.021 g/cm(2), NS) was not different in the two groups. Stepwise regression analysis showed significant correlations between BMD and race at each skeletal site except the lumbar spine and trochanter. Deconvolution analysis revealed no racial difference in any of the GH measurements. Whereas serum testosterone, serum 17 beta estradiol, and serum IGF-I were not different, serum IGFBP-3 was higher and the molar ratio of serum IGF-l/IGFBP-3 was lower in white than in black males. In summary, prepubertal BMD is higher in black than in white males at the hip, trochanter, and femoral neck, and the racial difference does not result from differences in secretion of GH. PMID- 11907711 TI - Effects of age and menopause on spinal bone mineral density in Japanese women: a ten-year prospective study. AB - In order to determine the age and menopause-related changes in spinal bone mineral density (BMD) in healthy Japanese women, the spinal BMD at L(2-4) was measured by dual X-ray absorptiometry (DXA) in 172 healthy Japanese women aged 31 69 years (mean age 53.1+/-6.7 years) in 1990 and 2000. This prospective study showed that there was no significant decrease of BMD in premenopausal women, but there was a significant decrease of BMD (-1.59%/year) in the early post menopausal women when compared with the premenopausal and late postmenopausal women (P <0.0001). The rate of decrease in BMD slowed down with the prolongation of the years since menopause (YSM). In postmenopausal women the annual rate of decrease in BMD for obese women was significantly lower than that for slim ones (P <0.01), suggesting that fat tissue may be effective for preventing bone loss. A multiple regression analysis of variables contributing to the annual rate of decrease in spinal BMD showed that YSM and physiological age were the most influential factors, considering other factors such as weight, height and bone mass index. In conclusion, an accelerated bone loss was seen in the early postmenopausal stage. The YSM and physiological age were the most important factors that affect the rate of bone loss in healthy postmenopausal Japanese women. PMID- 11907712 TI - COLIA1 Sp1 polymorphism predicts response of femoral neck bone density to cyclical etidronate therapy. AB - Genetic factors are important in the pathogenesis of osteoporosis but less is known about their possible role in predicting response to anti-osteoporotic therapy. Previous studies have shown that a polymorphic Sp1 binding site in the collagen type 1 alpha 1 gene (COLIA1) is associated with bone mineral density (BMD) and osteoporotic vertebral fracture. In this study we sought to determine if the COLIA1 Sp1 polymorphism might also act as a predictor of the response to treatment of osteoporosis with bisphosphonate therapy. The study group comprised 108 perimenopausal women with osteopenia who had been randomized to receive cyclical etidronate therapy for 2 years with a 1-year treatment-free follow-up as part of a randomized placebo controlled trial. Bone mineral density was measured at the lumbar spine and femoral neck by dual X-ray absorptiometry and genotyping performed on DNA extracted from peripheral blood leukocytes using standard techniques. The distribution of COLIA1 genotypes was similar to that previously reported in Caucasians with 69 (63.9%) "SS" homozygotes, 38 (35.2%) "Ss" heterozygotes, and 1 (0.9%) "ss" homozygote. There was no association between COLIA1 genotype and response of lumbar spine BMD during etidronate treatment or the follow-up phase. The response of femoral neck (FN) BMD, however, differed significantly between the genotype groups throughout the study period, such that FN BMD increased by 0.56%, 2.36%, 1.82%, and 1.32 % after 1, 2, 2.5, and 3 years, respectively in the "SS" genotype group, compared with -1.56%, -0.62%, -0.37%, and -0.66% in the "Ss/ss" genotype groups (P = 0.002). The data presented here show that site-specific heterogeneity exists in the response of BMD to cyclical etidronate therapy, which is related to COLIA1 genotype. Our data raise the possibility that COLIA1 genotyping could be used to target etidronate therapy to those most likely to respond in terms of FN BMD, with potential benefits in terms of economic cost and clinical outcome. PMID- 11907713 TI - Precision and accuracy of a transportable dual-energy X-ray absorptiometry unit for bone mineral measurements in guinea pigs. AB - Dual energy X-ray absorptiometry (DXA) is a valuable tool for measuring bone mineral content (BMC) and bone mineral density (BMD) in small-animal research. The present study was devised to establish guidelines and to define sites for bone mineral measurements in guinea pigs and to evaluate the accuracy of a new transportable research DXA unit. Repeated scans were performed on 30 guinea pig hindlimbs (in situ) as well as the isolated bones from these limbs (ex situ). Nine exactly specified regions of interest (ROIs) were analyzed twice for BMC and BMD by three different observers. Additionally, the BMC of whole bones and bone segments as measured by DXA was correlated to ash weights of bone in a subset of five animals to determine the accuracy of the DXA measurements. On ex situ scans, intra-observer variability for BMD ranged from 0.09% to 2.33% and inter-observer variability from 0.23% to 5.86% depending on the site studied, with smaller ROIs exhibiting more variability. Coefficients of variance (CV) for BMC measurements were slightly higher than for BMD. However, BMC offered a better correlation between in situ and ex situ values than BMD. On in situ scans, observer variability for BMD and BMC for comparable sites was higher than the ex situ variability. The results of this study indicate that DXA provides an accurate measurement of BMC even in small specimens. The precision of BMC and BMD measurements in situ can be improved considerably by using specific, well-defined ROIs and by careful placement of the bones to be scanned in close proximity to the scanning surface. PMID- 11907715 TI - Rat hindlimb unloading by tail suspension reduces osteoblast differentiation, induces IL-6 secretion, and increases bone resorption in ex vivo cultures. AB - In this research we utilized tail-suspended rats as an in vivo model for bone loss studies in order to investigate the effects of the tail suspension on the structure of the suspended bones and in ex vivo cultures the activities of trabecular osteoblasts, marrow-derived osteogenic cells, and osteoclasts obtained from treated animals, compared with untreated controls. After a 5-day hind limb unloading, trabecular thinning was already evidenced in the tibial primary spongiosa. In the secondary spongiosa, the bone formation activity was reduced whereas osteoclastic parameters were not yet altered. Bone marrow-derived osteogenic cells and differentiated osteoblasts from enzymatic digestion of posterior limb trabecular bone were prepared from 5 day tail-suspended rats and from normally loaded rats as controls. Cell morphology, alkaline phosphatase (ALPH) activity, production of mineral matrix, osteocalcin, and IL-6 secretion were evaluated in both cell populations. Tail suspension reduced the osteogenic potential of stromal marrow cells and of already differentiated osteoblasts. In fact, ALP positive colonies were significantly reduced in number and were smaller in size compared with controls and bone nodules formed in permissive conditions were also significantly fewer and smaller, whereas in cultures of cells from control conditions, large mineralizing nodules were formed. Osteocalcin secretion was not affected by unloading. Finally, IL-6 concentration was increased in marrow-derived cells from treated rats compared with controls. Primary cultures of osteoclasts were obtained from the nonadherent fraction of the bone marrow of the same animals. The number of TRAP positive cells in culture from tail suspended rats was significantly increased, as well as bone resorption activity, measured as resorbed surfaces of a suitable synthetic hydroxyapatite, compared with controls. These data clearly suggest that skeletal unloading not only reduces the osteogenic potential of osteoblastic cells but induces an increased osteoclastogenesis and osteoclast activity in ex vivo cultures. They also indicate for the first time that a possible mediator responsible for the increased osteoclastogenesis could be represented by the IL-6 whose secretion by bone marrow cells was significantly enhanced by unloading. PMID- 11907714 TI - The estrogen receptor ligand ICI 182,780 does not impair the bone-sparing effects of testosterone in the young orchidectomized rat model. AB - Testosterone (T) can affect bone metabolism not only directly, but also via its metabolites, estrogen or dihydrotestosterone, produced by enzymes present in bone. Therefore, the aim of this study was to investigate whether the high affinity estrogen receptor ligand ICI 182,780 (ICI) impaired the bone-protective action of T in 3-month-old orchidectomized (Orch) rats, studied during an experimental period of 3 months. As expected, Orch significantly decreased trabecular bone volume in the proximal tibial metaphysis (-52%), as measured by histomorphometry, and had a similar negative effect on volumetric bone mineral density (BMD) in the distal femoral metaphysis (-53%), as assessed by peripheral quantitative computed tomography (pQCT). The loss of bone induced by Orch was completely prevented by T administration. Moreover, the Orch-associated increases of biochemical markers of bone turnover (serum osteocalcin, urinary deoxypyridinoline, and calcium excretion) did not occur when Orch rats received T. Administration of ICI in combination with T did not impair this bone-sparing effect. Cortical bone parameters (as determined by pQCT), body weight gain, and body composition (as measured by dual-energy X-ray absorptiometry) were not affected by T or ICI in combination with T. Furthermore, no differences were observed in serum concentrations of insulin-like growth factor-I or glucose homeostasis. In conclusion, ICI does not impair the long-term bone-protective effects of T in orchidectomized male rats, suggesting that testosterone can mediate its effect on the male skeleton directly via the androgen receptor. The absence of effects on body growth via the growth hormone--insulin-like growth factor-I axis may be a possible explanation for the lack of skeletal effects of this selective estrogen receptor antagonist. PMID- 11907716 TI - Biological variability in serum and urinary indices of bone formation and resorption in dogs. AB - Serum and urinary assays of bone markers provide a noninvasive alternative to bone biopsy in the study of bone metabolism in humans. Many of the commercial assays that were originally developed for use in humans have been shown to cross react in dogs, and it should therefore be possible to use these assays to study bone remodeling in dogs. The interpretation of bone marker data in humans is hampered by extensive inter- and intra-subject variability. The specific aim of this study was therefore to determine the extent of biological variability in bone markers in dogs. Serum and urine samples were collected every 4 hours over a 24-hour period (short-term study), and every week over a 12-week period (long term study). Serum bone-specific alkaline phosphatase (BALP) and urinary deoxypyridinoline (Dpd) and N-terminal telopeptide of collagen (NTx) were measured with commercial enzyme immunoassays. Serum osteocalcin (OC) and carboxyterminal crosslinked telopeptide of type I collagen (ICTP) were measured with commercial radioimmunoassays. In the short-term study, statistically significant diurnal rhythms were identified for OC, BALP, ICTP, and Dpd. No clear rhythm was evident for NTx. There was no evidence of statistically significant long-term variability in marker excretion over the 12 weeks. Our findings confirm the utility of these assays in dogs. However, as in humans, care must be taken to ensure that specimens are collected at a consistent time of day. Moreover, given the inherent variability in marker excretion in individual animals, the most appropriate use for these assays is as a screening tool for cohort studies, rather than as a diagnostic or prognostic tool in the individual animal. PMID- 11907717 TI - The NMDA type glutamate receptors expressed by primary rat osteoblasts have the same electrophysiological characteristics as neuronal receptors. AB - Cells of mammalian bone express glutamate receptors. Functional N-methyl-D aspartate (NMDA) receptors have been demonstrated in human, osteoblastic MG-63 cells, but currents in these cells, unlike those of mammalian neurons, are blocked by Mg(2+) in a voltage-insensitive manner. Differences between the characteristics of NMDA currents in bone cells and in neurons may reflect molecular variation of the receptors or associated molecules, with implications for the role(s) of glutamate in these different tissues and for targeting of ligands/antagonists. To determine whether NMDA receptors in primary bone cells are functional, and whether the currents carried by these receptors resemble those of MG-63 cells or those of mammalian neurons, we have applied the whole cell patch clamp technique to primary cultures of rat osteoblasts. In 0-Mg(2+) saline, 25% of cells showed a slowly developing inward current in response to bath perfusion with 1 mM or 100 microM NMDA. Antibodies against NMDA receptors stained approximately 26% of cells. When NMDA was applied by rapid superfusion, kinetics of the currents were similar to those of neuronal NMDA currents, reaching a peak within 20-30 ms. 1 mM Mg(2+) reduced current amplitude at negative holding potentials and caused the I-V relationship of the currents to adopt a 'J' shape rather than the linear relationship seen in the absence of added Mg(2+). Co-application of glycine (20 microM) with NMDA increased current amplitude by only 18%, suggesting that glycine is released from cells within the cultures. Currents were blocked by (+)-MK-801 and DL-2-amino-5-phosphonovaleric acid. Fluorimetric monitoring of [Ca(2+)](i) using fura-2 showed that, in Mg(2+) free medium, NMDA caused a sustained rise in [Ca(2+)](i) that could be reversed by subsequent application of MK-801. We conclude that rat femoral osteoblasts express functional NMDA receptors and that these receptors differ from those previously identified in MG-63 cells. NMDA receptors of primary osteoblasts show a 'classical' voltage-sensitive Mg(2+) block, similar to that seen in neuronal NMDA receptors, and will therefore function as detectors of coincident receptor activation and membrane depolarization. PMID- 11907718 TI - Prophylaxis of rebleeding from isolated gastric fundal varices by balloon occluded retrograde transvenous obliteration. AB - We describe a case of bleeding gastric fundal varices associated with a gastrorenal shunt that were successfully treated with balloon-occluded retrograde transvenous obliteration. Blood flow in the varices disappeared after treatment. Because of its safety and simplicity, balloon-occluded retrograde transvenous obliteration appears to be a feasible alternative to transjugular intrahepatic portosystemic shunt for the treatment of gastric fundal varices. PMID- 11907719 TI - Ruptured gastric fundal varices treated with endoscopic variceal ligation and transjugular retrograde obliteration: case report. AB - Therapeutic modalities for the obliteration of collateral vessels connecting the portal venous system with the systemic circulation, transjugular retrograde obliteration (TJO) and balloon-occluded retrograde transvenous obliteration have recently been developed, and several satisfactory results have been reported with their use. We report a case of ruptured gastric fundal varices treated with TJO after endoscopic variceal ligation (EVL). In our case, variceal bleeding was controlled successfully with EVL and varices were eradicated with TJO. PMID- 11907720 TI - Prolapsed hyperplastic gastric polyp causing pancreatitis: case report. AB - A huge hyperplastic gastric polyp prolapsed into the duodenum. The compression and obstruction of the ampulla of Vater by this polyp caused acute pancreatitis. An overview of imaging findings, general considerations about hyperplastic gastric polyps, and a review of the literature are provided. PMID- 11907721 TI - Barium trapping in rectoceles: are we trapped by the wrong definition? AB - BACKGROUND: Barium trapping within a rectocele is a criterion used by surgeons to select which patients with rectoceles should undergo operative repair. This proctographic study compared the presence and depth of barium trapping within a rectocele on postevacuation radiography with those seen on posttoilet radiography after further evacuation in the privacy of the bathroom. METHODS: Eighty-two consecutive patients with evidence of barium trapping on postevacuation radiographs of a fluoroscopic dynamic cystoproctographic examination were reviewed retrospectively. The size of the rectoceles and the depth of barium trapping on the postevacuation and subsequent posttoilet radiographs were measured. RESULTS: The posttoilet radiographs showed resolution of the barium trapping in 47 (57%) of the 82 patients. Resolution of the trapping was directly related to rectocele size. The mean differences in the depth of barium trapping between the postevacuation and posttoilet radiographs were significant for all sizes of rectocele. CONCLUSION: Barium trapping in rectoceles changes with the degree of rectal evacuation. More complete evacuation was shown on the posttoilet radiograph than on the postevacuation radiograph. Consequently, the posttoilet radiograph may be more appropriate for the preoperative assessment of barium trapping within rectoceles. PMID- 11907722 TI - Per-rectal contrast material for abdominal CT: usefulness of administration with an automatic injector. AB - Adequate distention of the gastrointestinal tract is essential for the best quality image in abdominal computed tomography. We introduce a new technique for per-rectal administration of contrast material with the use of an automatic injector. With this technique, more contrast material can be inserted and thus adequately distend the colon, including the proximal colon. PMID- 11907723 TI - Ciliated hepatic foregut cyst: case report with an emphasis on US findings. AB - Ciliated hepatic foregut cyst (CHFC) is a rare mass, and its medical imaging findings are seldom reported. We present a histologically proven case of CHFC. The patient was asymptomatic, and a mass was found incidentally by sonography (US) in segment IV. The lesion was almost anechoic on fundamental US but filled with dense echoes with distinct posterior echo enhancement on second harmonic imaging. The lesion was avascular on color Doppler US and angiography. Thus, when US detects a mass in segment IV, the possibility of a CHFC should be considered regardless of the US results. PMID- 11907724 TI - Contrast-enhanced ultrasound improves hepatic vessel visualization after orthotopic liver transplantation. AB - BACKGROUND: We investigated whether color-coded Doppler sonography combined with an ultrasound contrast medium would improve the assessment of liver-supplying vessels after orthotopic liver transplantation. METHODS: Forty-seven patients after orthotopic liver transplantation participated. Examinations were done without and then with the ultrasound contrast medium Levovist. Visualization of the liver-supplying vessels was assessed with a scoring system. RESULTS: Visualization of the portal vein was similar without and with contrast medium. Hepatic arteries were visualized in 39 patients without contrast medium and 46 patients with contrast medium. The remaining patient showed hepatic artery thrombosis, which was confirmed angiographically. With the use of Levovist, the examination took 3.7 min rather than the usual 6.4 min. CONCLUSION: Imaging of hepatic arteries after liver transplantation improved significantly with the use of ultrasound contrast medium. These findings are important because the early detection of blood flow through the liver after transplantation affects prognosis. PMID- 11907725 TI - Cirrhosis: spectrum of findings on unenhanced and dynamic gadolinium-enhanced MR imaging. AB - The appearance of the cirrhotic liver on computed tomography can be difficult to evaluate and can frustrate the radiologist distinguishing benign from malignant lesions. Hepatic edema, fibrosis, atrophy, and vascular abnormalities are common in the cirrhotic liver and produce derangements in morphology, attenuation, and perfusion, limiting the accurate characterization of hepatic masses. With the development of fast magnetic resonance (MR) sequences and dynamic postgadolinium enhanced imaging, most hepatic lesions with uncertain etiology on computed tomography can be accurately characterized on MR imaging. We describe MR imaging techniques useful for imaging cirrhosis and its complications. We also illustrate the spectrum of findings in the cirrhotic liver on dynamic gadolinium-enhanced MR imaging, including reticular and confluent fibrosis, fatty infiltration, hemochromatosis, regenerating nodules, dysplastic nodules, hepatocellular carcinoma, and sequela of portal hypertension. PMID- 11907726 TI - Extrahepatic arterioportal venous fistula: multidetector CT and volume-rendered angiographic imaging. AB - We present a case of unsuspected extrahepatic arterial-main portal venous fistula diagnosed by multiphase computed tomography with three-dimensional volume rendered computed tomographic angiography. To our knowledge, this entity has not been reported previously in the literature, and it represents an example of pathology that may only be detected and clarified with multiphase imaging with three-dimensional rendering. This finding has great clinical importance in patient management. PMID- 11907727 TI - Localized gallbladder carcinoma: sonographic findings. AB - Our study of color (seven cases) and contrast (three cases) Doppler results of seven cases with gallbladder carcinoma localized in the gallbladder wall (TNM stage T1) showed that the presence or absence of blood flow signals distinguishes gallbladder carcinoma in stage T1b (muscular involvement) from tumefactive biliary sludge and that injection of contrast medium markedly increased diagnostic confidence. Thus, when color Doppler sonography is ambiguous, contrast enhanced Doppler sonography is the next line of investigation. However, actual color Doppler sonography is still not fully capable of displaying fine blood flow signals from gallbladder carcinoma in stage T1a (mucosal involvement), and greater Doppler sensitivity is mandatory for this purpose. PMID- 11907728 TI - Utility and limitations of intraductal ultrasonography in distinguishing longitudinal cancer extension along the bile duct from inflammatory wall thickening. AB - BACKGROUND: We wanted to distinguish wall thickening caused by cancer extension from that caused by inflammation after placing a biliary catheter on intraductal ultrasonography (IDUS). METHODS: We studied 51 patients with biliary tract malignancies who had undergone placement of biliary drainage catheters before IDUS. IDUS was performed from a transhepatic (n = 34) or transpapillary (n = 17) route with a thin-caliber ultrasonic probe (2.0 mm in diameter, 20-MHz frequency). At the hepatic side of the tumor, the thickness, asymmetry, outer margin, inner margin, and internal echoes of the bile duct wall were reviewed prospectively and correlated with the histologic findings of the surgically resected specimens in all cases. RESULTS: When IDUS showed wall thickening in a semicircular fashion, notched outer margin, rigid inner margin, papillary inner margin, and heterogeneous internal echoes, each finding had a positive predictive value for diagnosing cancer extension (100%, 100%, 83%, 100%, and 90%, respectively). When these factors were used as the diagnostic criteria of cancer extension, IDUS accurately demonstrated suitable surgical margins in 76% of all patients and 71% of patients with bile duct carcinoma. CONCLUSION: Wall thickening in a semicircular fashion, notched outer margin, rigid or papillary inner margin, and heterogeneous internal echoes are specific for cancer extension. However, surgical margins can be inaccurately assessed in some patients. PMID- 11907729 TI - Intraductal ultrasonography of traumatic neuroma of the bile duct. AB - We report a case of a 70-year-old female with traumatic neuroma of the bile duct. Transpapillary intraductal ultrasonography showed a cystic duct stump, from which a smooth and homogeneous hypoechoic mass arose; the adjacent bile duct wall had a normal structure. Intraductal ultrasonography is useful for distinguishing traumatic neuroma from bile duct carcinoma. PMID- 11907730 TI - Late-phase enhancement of the upstream portion of pancreatic adenocarcinoma on dual-phase helical CT. AB - BACKGROUND: Late-phase enhancement of pancreatic parenchyma upstream (tail side) of pancreatic adenocarcinoma is found frequently on dual-phase helical computed tomography (CT). We measured the frequency of late-phase enhancement of the upstream portion of pancreatic adenocarcinoma and normal pancreatic parenchyma using dual-phase helical CT. METHODS: Twenty-one patients with pancreatic adenocarcinoma and nontumorous pancreas upstream of tumors were compared with 100 control patients without pancreatic disease. Early and late scans started at 25 and 75 s, respectively, after intravenous injection of contrast material. The attenuation values of normal and nontumorous pancreas upstream of tumors were assessed in three phases: precontrast, early, and late enhanced. Enhancement ratio (ER) was calculated as ER = (late phase - precontrast)/(early phase - precontrast). RESULTS: Late-phase enhancements (ER > 1.0) were seen in 86% of upstream pancreas and 10% of normal pancreas. The mean ER of upstream pancreas was significantly higher than that of normal pancreas (p < 0.01). CONCLUSION: Late-phase enhancement of the pancreas upstream of the tumor is frequently observed in patients with pancreatic adenocarcinoma. Late-phase enhancement and histology showed a correlation for chronic obstructing pancreatitis in five patients. PMID- 11907731 TI - Cystic pancreatic masses: cross-sectional imaging observations and serial follow up. AB - BACKGROUND: We retrospectively reviewed the imaging features of a series of patients with cystic pancreatic masses, the majority of whom underwent imaging surveillance. METHODS: Imaging data from 30 patients with known cystic pancreatic masses were reviewed. Nine patients had surgical and/or cytologic classification. Of the 21 who were not operated on, all underwent serial imaging surveillance. Of these, five had corroborative endoscopic retrograde cholangiopancreatography and 16 were followed by only computed tomography and/or magnetic resonance imaging. RESULTS: In the nonoperated group, mean follow-up time was 30 months (3-144 months). Two patients demonstrated growth, and the remainder remain stable. In the patients who underwent surgery, invasive carcinoma was found in those with lesions larger than 4 cm, involvement of the main pancreatic duct, or visible solid components on the imaging study. Smaller lesions were benign. CONCLUSION: In patients with suspected cystic pancreatic neoplasms, surveillance might be possible if lesions are smaller than 2.5 cm, spare the main pancreatic duct, and demonstrate no solid components. PMID- 11907732 TI - Pancreatic cystosis in cystic fibrosis: case report. AB - We describe a 19-year-old cystic fibrosis patient, with pancreatic insufficiency since the age of 4 who presented at the age of 13 with postprandial abdominal pain. Ultrasonography and computed tomography showed several pancreatic cysts that progressively increased in diameter over 6 years. The lack of association with clinical and biochemical signs of acute pancreatitis is highlighted. PMID- 11907733 TI - Successful treatment of a ruptured mycotic aneurysm of the ileocolic artery with transcatheter embolization and antibiotic therapy. AB - We report a case of mycotic aneurysm of the ileocolic artery due to Streptococcus bovis endocarditis and acute septicemia complicated by active hemorrhage, that was treated successfully with transcatheter embolization and subsequent intravenous antibiotic treatment. This case suggests that a mycotic aneurysm can be treated successfully by percutaneous embolization in an emergent situation (active bleeding, septicemia) even without previous antibiotic therapy. PMID- 11907735 TI - Renal arteriovenous malformation: sonographic findings. AB - BACKGROUND AND METHODS: Renal arteriovenous malformation (RAVM) is a relatively rare congenital disease. Although sonography (US) currently is the first diagnostic tool for examining the kidney, its US and color Doppler findings have seldom been reported. We reviewed the clinical manifestations and US results of five cases of RAVM to clarify the role and limitations of US in the diagnosis. RESULTS: The lesions were solitary in all cases, and the affected side was the right in four cases and the left in one case. In four cases, the patients complained of hematuria, but the remaining case had no symptoms. US did not detect the lesion, but in all cases color Doppler US showed a focal vascular lesion with posterior color spots. US reexamination with knowledge of the Doppler results did not show any focal lesion. CONCLUSION: US was not diagnostic for RAVM, and color Doppler US should be performed immediately in patients with hematuria. PMID- 11907734 TI - Usefulness of power Doppler and contrast-enhanced sonography in the differentiation of hyperechoic renal masses. AB - BACKGROUND: In a prospective study, we compared power Doppler with and without contrast medium in the depiction of vascularity for the characterization of hyperechoic renal lesions. METHODS: Forty-one hyperechoic renal expansive lesions (29 benign, 12 malignant) in 32 patients were studied with power-Doppler ultrasonography before and after administration of an echo-enhancing agent (Levovist Schering AG, Berlin, Germany). Vascular architecture of the lesions was categorized into five different patterns. RESULTS: Power Doppler ultrasonography showed vascular structures in 25 lesions. The study enhanced with Levovist showed vascularity in eight of 16 lesions not seen on the unenhanced study. The characterization of vascular patterns with unenhanced power Doppler ultrasonography improved diagnostic accuracy compared with gray-scale ultrasonography (59% vs. 32%). The combination of B mode and power Doppler produced even greater diagnostic accuracy (78%), independent of the administration of echo-enhancing agent. Levovist administration was useful in the differential diagnosis between pseudotumor and neoplasm. CONCLUSION: The use of sonographic contrast agent did not increase the diagnostic accuracy of power Doppler in the differential diagnosis of hyperechoic renal lesions but was advantageous for the characterization of suspected pseudomasses. PMID- 11907736 TI - Bladder cancer within a direct inguinal hernia: CT demonstration. AB - We present a case of a urothelial neoplasm arising within a direct bladder hernia in the inguinal canal. Bladder hernias are rarely found preoperatively and are exceptional sites of neoplasm. Spiral computed tomography with gaseous insufflation of the bladder demonstrated the bladder hernia and the extension of the neoplasm in the inguinal canal more accurately than other computed tomographic techniques with nonopacified and iodinated urine. PMID- 11907740 TI - GPIIb-IIIa receptor inhibitors: what the interventional radiologist needs to know. AB - The glycoprotein IIb-IIIa (GPIIb-IIIa) receptor inhibitors have established themselves as first line therapy in the treatment of acute coronary syndromes (ACS) and percutaneous coronary intervention (PCI). The benefit of these agents rests in their ability to attenuate the deleterious effects of platelet activation, both at the site of an inflamed vessel wall (due to a ruptured plaque or PCI) and in the microcirculation as a result of embolization. Based on these results, interventional radiologists are beginning to explore the potential of using GPIIb-IIIa inhibitors during interventions in the peripheral circulation. This paper reviews the molecular biology of the GPIIb-IIIa receptor, the pharmacology of the GPIIb-IIIa receptor inhibitors, the current coronary and peripheral vascular literature as it pertains to the GPIIb-IIIa receptor inhibitors, and potential future applications of the GPIIb-IIIa receptor inhibitors in the peripheral circulation. PMID- 11907741 TI - Intraarterial lidocaine administration for relief of pain resulting from transarterial chemoembolization of hepatocellular carcinoma: its effectiveness and optimal timing of administration. AB - PURPOSE: Patients undergoing transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) commonly have significant post-procedural abdominal pain necessitating narcotic administration. It is known that intraarterial administration of lidocaine is effective in controlling the pain during the procedure. However, optimum timing of the lidocaine administration is not precisely known. The purpose of this study was to assess the efficacy of intraarterial lidocaine administration for control of pain resulting from TACE and to evaluate the optimal timing of administration. METHODS: In a prospective trial, 113 consecutive patients with HCC who underwent TACE were classified into three groups: those who received a lidocaine bolus intraarterially immediately prior to TACE (group A, n = 30), those who received lidocaine immediately after TACE (group B, n = 46), and those who did not received lidocaine (group C, n = 37). Incidence and degree of post-procedural pain was assessed using a subjective method (visual analogue scales scored from 0 to 10) and an objective method (amount of post-procedural analgesics). RESULTS: The incidence of post-procedural pain in group A (16.7%) was significantly lower than that of group B (38.3%; p = 0.005). The mean pain score was 3.0 in group A and 4.8 and 3.1 in groups B and C, respectively. The mean dose of analgesic used after the procedure in group A (25.0 mg) was significantly lower than those in group B (52.9 mg) and group C (41.0 mg; p = 0.002). CONCLUSIONS: Pre-TACE intraarterial administration of lidocaine is much more effective than post-TACE administration in reducing the incidence and the severity of post-procedural pain. Furthermore, in order to reduce the incidence of post-procedural pain and dose of post-procedural analgesics, we recommend routine pre-TACE administration of lidocaine because post-procedural pain might developed even in patients who did not feel any pain during the TACE. PMID- 11907743 TI - Early results of retrograde transpopliteal angioplasty of iliofemoral lesions. AB - PURPOSE: To assess whether the retrograde transpopliteal approach is a safe, practical and effective alternative to femoral puncture for percutaneous transluminal angioplasty (PTA). METHODS: Forty PTAs in 38 patients were evaluated. Intentional subintimal recanalization was performed in 13 limbs. Ultrasound evaluation of the popliteal fossa was carried out 30 min and 24 hr postprocedurally in the first 10 patients to exclude local complications. All patients had a follow-up of at least 6 weeks. RESULTS: The indication for PTA was critical ischemia in seven limbs and disabling claudication in the remainder. Stenoses (single or multiple) were present in 24 and occlusion in 15. The superficial femoral artery (SFA) was the commonest segment affected (36) followed by common femoral artery (CFA) in four and iliac artery in four. Technical success was achieved in 38 of 39 limbs where angioplasty was carried out. In one limb no lesion was found. Immediate complications were distal embolization in two and thrombosis in one. None of these required immediate surgery. There were no puncture site hematomas or popliteal arteriovenous fistulae. Symptomatic patency at 6 weeks was 85%. Further reconstructive surgery was required in three limbs and amputation in two. CONCLUSION: The transpopliteal approach has a high technical success rate and a low complication rate with a potential to develop into an outpatient procedure. It should be considered for flush SFA occlusions or iliac disease with tandem CFA/SFA disease where the contralateral femoral approach is often technically difficult. PMID- 11907742 TI - Renal artery stenting in patients with a solitary functioning kidney. AB - PURPOSE: To retrospectively evaluate the results of renal artery stenting in patients with renovascular disease and a solitary functioning kidney. METHODS: Palmaz stents were placed in 16 patients with a solitary functioning kidney, renal artery stenosis, hypertension and renal failure. Stenoses were evaluated with color Doppler ultrasound, MR angiography and digital subtraction angiography (DSA). Indications for stenting were: recoil after percutaneous transluminal renal angioplasty (PTRA) (63%), arterial dissection after PTRA (13%) and primary stenting (25%). Immediate results were evaluated by DSA. On follow-up (6-36 months), patients underwent periodical evaluation of clinical conditions (blood pressure and serum creatinine level) and stent patency, by means of color Doppler ultrasound. RESULTS: Stent placement was successful in all patients (100%). Cumulative primary patency rate was: 100% at 1 day, 93.75% at 6 months, 81.25% at 12 months and 75% at 24 months. A significant reduction in diastolic blood pressure occurred (mean +/- SD 104 +/- 6 vs 92 +/- 3; p < 0.05); renal function improved or stabilized in over 80% of patients. However, there was no significant difference in the creatinine values before and after treatment (mean +/- SD 200 +/- 142 micromol/l vs 197 +/- 182 micromol/l; p > 0.05). CONCLUSION: Renal artery stenting, both after PTRA and as primary stenting, represents a safe procedure, able to preserve renal function in patients with a solitary functioning kidney. PMID- 11907744 TI - Accumulation of iodized oil within the nonneoplastic liver adjacent to hepatocellular carcinoma via the drainage routes of the tumor after transcatheter arterial embolization. AB - PURPOSE: After transcatheter arterial embolization (TAE) with iodized oil (Lipiodol), a relatively dense accumulation of Lipiodol is often seen in the nontumorous liver adjacent to a hypervascular hepatocellular carcinoma (HCC) nodule. We compared this phenomenon with the findings obtained with single-level dynamic CT during hepatic arteriography (SLDCTHA) and presumed its possible mechanism. METHODS: Fifty-six patients with HCC underwent hepatic angiography including SLDCTHA followed by segmental or subsegmental TAE with a mixture of an anticancer drug and Lipiodol. We compared the drainage area of the HCC depicted on SLDCTHA with the Lipiodol accumulation in the nontumorous liver adjacent to the HCC on CT after TAE (LpCT). RESULTS: In 26 of the 56 patients, a definite corona enhancement around the HCC, suggesting the drainage of blood from the tumor into the surrounding liver parenchyma, was seen on the late phase of SLDCTHA. In 17 of these 26 patients (65.4%), LpCT showed a more intense accumulation of Lipiodol in the nontumorous liver adjacent to the HCC that corresponded to the drainage area revealed on SLDCTHA. CONCLUSION: The drainage of blood from the HCC was considered to be a possible mechanism of the accumulation of Lipiodol in the nontumorous liver adjacent to the HCC. PMID- 11907745 TI - Long-term results of endovascular stent placement in the superior caval venous system. AB - PURPOSE: To present the long-term results in superior caval stenting for symptomatic obstruction. METHODS: Forty-nine stents were placed in 30 patients: 16 (53%) with malignant lesions, five (17%) with benign lesions and nine (30%) hemodialysis patients. Self-expandable stents were deployed on a first-line basis. Patients were followed clinically as well as by various imaging techniques and survival analysis was performed. RESULTS: Stent deployment was possible in all cases. Reocclusion was seen in 13 patients, of whom eight belonged to the hemodialysis group. Primary and secondary patency rates for malignant, benign and hemodialysis patients were respectively 74%, 50% and 22%, and 74%, 75% and 56% at 1 year. We had 7% complications and one death from iatrogenic superior vena cava injury. CONCLUSION: Primary stenting of superior caval obstruction is a first choice treatment method achieving good mid-term patency. Patients with hemodialysis shunts must be closely monitored for early reintervention. PMID- 11907746 TI - Radiologic placement of a low profile implantable venous access port in a pediatric population. AB - PURPOSE: To evaluate the feasibility and complications of placement of a low profile venous access port in the chest in children requiring long-term venous access. METHOD: A low-profile peripheral arm port (PAS port; Sims Deltec, St. Paul, MN, USA) was implanted in the chest in 22 children over a 4-year period. The mean age of the study group was 6 years (range: 9 months to 20 years). Ports were placed for the administration of chemotherapy, hyperalimentation and frequent blood sampling. Sonographic guidance was used to access the internal jugular or subclavian vein in each case. A review of all inpatient and outpatient charts was undertaken to assess catheter performance and complications. RESULTS: Access to the central venous circulation was successfully achieved in each case without complication. Ports remained implanted for 6579 catheter-days (mean: 299 days). Ten ports have been removed. Of three patients (13%) experiencing device related infections (0.45 infections/1000 catheter days), two (9.1%) were unresponsive to antibiotics and removed (0.3 infections/1000 catheter days). One port was removed because of pain in the shoulder adjacent to the port implantation site. One port was removed because of difficult access. The final port was removed in order to place a dual-lumen catheter prior to bone marrow transplant. Twelve ports remain implanted. Aspiration occlusion occurred in four patients (18%). Deep venous thrombosis did not occur in any patient. CONCLUSION: Low-profile chest ports placed by interventional radiologists in the interventional radiology suite can be placed in children as safely as traditional chest ports placed in the operating room. The incidence of infection, venous thrombosis and aspiration occlusion is comparable to that of ports placed operatively. PMID- 11907747 TI - Prevention of recurrent central venous stenosis using endovascular irradiation following stent placement in hemodialysis patients. AB - This study was done to evaluate the outcome after brachytherapy (BT) given to prevent restenosis after stent insertion for central venous stenosis in patients with ipsilateral hemodialysis arteriovenous fistulas (AVF). Angioplasty and stenting were performed on 9 primary central venous stenoses in 8 patients with AVF followed by BT, delivering Iridium-192 radiation using an afterloading technique. BT was also administered to three patients with five recurrent stenoses at the stent margins. There was no residual stenosis after angioplasty and stenting. Venographic follow-up (77-644 days, mean 272 days) showed no restenosis in seven primary stenoses. New strictures (45%-100%) developed at the stent margin in six veins (five patients). Angioplasty or stenting was performed for five margin stenoses in three patients, followed by a second BT. Residual stenosis before BT was 0-30%. In our venographic follow-up (140-329 days, mean 215 days), three restenoses occurred (35%-100%). All progressed to complete occlusion on later venographic follow-up irrespective of whether BT was given to the stent margin or not. The mean primary and assisted primary patency of the central veins were 359 days and 639 days, respectively. Endovascular irradiation with a noncentering source does not prolong the patency after angioplasty and stenting of central venous stenosis in hemodialysis patients. PMID- 11907749 TI - Animal experience in the Gunther Tulip retrievable inferior vena cava filter. AB - PURPOSE: To assess the retrievability of the Gunther Tulip temporary inferior vena cava filter from a technical viewpoint, and consider the histopathologic changes that occur at the anchoring site of the filter prongs to the vein endothelium in Landrace pigs. METHODS: Twenty-two Gunther Tulip retrievable filters were inserted in 22 experimental Landrace pigs via the jugular vein. Device implantation time was 0, 3, 7, 12, 14, 15, 16, 20, 30, 35 and 56 days. Study subjects were divided into two groups. In one group the filter was retrieved percutaneously via the jugular vein whereas in the other group it was removed surgically. The specimens obtained (vena cava and filter) were histopathologically examined. Prior to filter retrieval, a venacavography was obtained in all cases. Degree of retrieval difficulty was rated as follows: no difficulty (N), slight (S), mild (M), high (H) and unretrievable (U). RESULTS: Of the 22 implanted filters, 11 should have been removed percutaneously but this was impossible in three cases (U). In four cases the device was retrieved with no difficulty (N); in two cases the degree of difficulty was mild (M) and in other two it was high (H) and slight (S) respectively. Retrieval difficulties were observed after 16 days. Starting from day 20, there was evidence of fibrosis with thick intimal proliferation and total filter prong involvement, which accounts for the difficulty in retrieving the device. CONCLUSIONS: It is advisable not to exceed a filter retrieval time of 16 days in view of the fibrotic changes reported. It might be necessary to perform a larger study with more animals and with retrieval times between 14 and 20 days. PMID- 11907748 TI - Microbubble potentiated ultrasound as a method of declotting thrombosed dialysis grafts: experimental study in dogs. AB - Intravenous perfluorocarbon-exposed sonicated dextrose albumin (PESDA) microbubbles in the presence of low frequency ultrasound (LFUS) can lyse very small clots. We develop a similar method to declot full-size arteriovenous dialysis grafts. Dialysis grafts fashioned in three dogs were cannulated and ligated. After thrombosis, three declotting techniques were randomly applied: 1) direct injection of PESDA + LFUS; 2) direct injection of saline + LFUS; and 3) intravenous PESDA + LFUS. Declotting was graded by cine angiography scores of each third of the graft on a scale of 0-4 (maximum total score = 12). Twenty-six procedures showed mean patency scores of 11.1 for direct PESDA and 8.4 for i.v. PESDA, vs 4.9 for direct saline, p = <0.001. All eight direct PESDA injections achieved lysis and good flow, but none of 8 direct saline injections succeeded, p = <0.01. Intravenous PESDA succeeded in 4 of 10 procedures, p = <0.04 vs saline. Direct injection of PESDA with transcutaneous LFUS succeeds in lysing moderate size clots and recanalizing thrombosed fistulas. PMID- 11907750 TI - Percutaneous retrieval of the Tulip vena cava filter: feasibility, short- and long-term changes--an experimental study in dogs. AB - PURPOSE: To evaluate experimentally the retrievability of the Tulip inferior vena cava (IVC) filter in an in vivo study. Changes which accompany venous healing after filter retrieval were investigated. METHODS: In 12 dogs, 23 filters were inserted percutaneously into the lumbar and intrahepatic segments of the IVC. Two weeks (n = 21 filters) or 3 weeks (n = 2 filters) after insertion, filter retrieval was attempted through an 11 Fr coaxial retrieval sheath system placed via the jugular vein. Follow-up studies before and after filter retrieval included cavography, computed tomography and intravascular ultrasound of the IVC. Seven dogs were killed immediately after filter retrieval to confirm short-term changes of the IVC, and 5 dogs were killed 6 months after filter retraction to evaluate long-term changes of the IVC related to filter retrieval. Post-mortem examinations and histologic specimens of the IVC were obtained to evaluate caval wall abnormalities secondary to filter removal. RESULTS: All but one filter were successfully retrieved 2 weeks post-implantation. However, 3 weeks after insertion, filter retrieval was impossible. There were no complications caused by filter extraction. Follow-up studies after filter retrieval revealed no significant changes in the integrity, morphology or composition of the IVC and pericaval tissue. Histologic examination 6 months after filter retrieval revealed only flimsy fibrotic intimal plaques at the sites of former hook insertion. CONCLUSION: The Tulip filter allows percutaneous insertion and retrieval up to 14 days after insertion, suggesting that it may be useful for either permanent or temporary prophylaxis against pulmonary embolism. PMID- 11907752 TI - Relentless progression of venous obstruction in a case of Budd-Chiari syndrome related to heterozygous protein C deficiency. AB - A 28-year-old man with heterozygous protein C deficiency presented with Budd Chiari syndrome resulting from hepatic vein obstruction. Over the next 40 months, standard oral anticoagulant therapy and multiple percutaneous interventions aimed at relieving hepatic vein obstruction could not prevent progression of the disease ultimately to cirrhosis and death. Serial angiography provided unique documentation of the relentless progression of hepatic venous obstruction, which was related to the disease and to iatrogenic factors. Operative findings obtained during unsuccessful mesocaval shunt surgery revealed that venous disease in protein C deficiency can be far more extensive than is clinically anticipated. The ineffectiveness of therapy in this patient may be related to standard oral anticoagulant therapy being insufficient to offset the risk of recurrent thrombosis and progression to an advanced stage of vascular damage. PMID- 11907751 TI - Oily chemoembolization of pancreatic head adenocarcinoma. AB - Oily chemoembolization is a known method of treatment for hepatic malignancies but was never used for pancreatic cancer. We report the case of a 48-year-old patient with unresectable adenocarcinoma of the pancreatic head treated by repeated chemoembolizations of the feeding (gastroduodenal) artery with gemcitabine-in-lipiodol. After 10 procedures there was a marked decrease in tumor. Radiologic examinations showed no signs of viable tumor. The patient is alive and symptom-free 22 months after the start of treatment. Oily chemoembolization should be investigated as a technically simple, safe, and potentially effective palliative management of unresectable pancreatic head carcinoma. PMID- 11907753 TI - MR-guided laser-induced thermotherapy of the infratemporal fossa and orbit in malignant chondrosarcoma via a modified technique. AB - A 76-year-old patient presented with a recurrent mass of a malignant chondrosarcoma in the right infratemporal fossa and in the left maxillary sinus with orbital invasion. The patient was treated with a palliative intention with MR-guided laser-induced thermotherapy using a modified applicator technique. Following treatment clinical symptoms improved and MRI revealed complete laser induced tumor necrosis. PMID- 11907754 TI - Extrahepatic portal vein tear with intraperitoneal hemorrhage during TIPS. AB - Temporary occlusion with an angioplasty balloon can stop intraperitoneal bleeding. PMID- 11907755 TI - Palliative treatment of an enterorectal fistula with a covered metallic stent. AB - In patients with recurrent colorectal cancer, the development of fistulae presents a difficult therapeutic problem. The traditional surgical approach of resection and intestinal diversion may not always be appropriate in those with disseminated or terminal disease. We present the successful use of a covered esophageal stent to occlude an enterorectal rectal fistula in a patient with recurrent inoperable colonic carcinoma. PMID- 11907756 TI - Recurrence of dialysis shunt pseudoaneurysm following percutaneous thrombin embolization. AB - We report an unusual case of a 58-year-old woman on maintenance hemodialysis who presented with an iatrogenic radial artery pseudoaneurysm proximal to a Cimino Brescia fistula. The pseudoaneurysm was not amenable to ultrasonographic compression due to the vascular anatomy. The pseudoaneurysm recurred despite initial successful response to two direct injections of thrombin on separate occasions. We highlight the role of ultrasound and thrombin injection in the treatment of pseudoaneurysm and emphasize the need for follow-up ultrasound examination to monitor recurrence. PMID- 11907757 TI - Fatal hemorrhage in stented esophageal carcinoma: tumor necrosis of the aorta. AB - We report a case of fatal hematemesis in a patient with an esophageal stent for palliation of malignant dysphagia. Post-mortem examination showed tumor thrombi in the vasa vasorum of the aorta causing aortic wall necrosis and perforation. Massive delayed hemorrhage is reported as a complication of metallic esophageal endoprostheses but in the majority of cases there have been no post-mortem examinations. In this case the post-mortem examination demonstrated that the cause of death was not stent-related. We feel that this case report adds important information to the current literature regarding the delayed complications seen in patients with esophageal carcinoma who have been palliated with metallic stents. PMID- 11907758 TI - Re: An alternative superior ophthalmic vein approach for transvenous embolization of dural arteriovenous fistulas of the cavernous sinus. PMID- 11907759 TI - Re: Treatment of a chronically draining infected hepatic echinococcal cyst cavity with repeated injection of 10% povidone-iodine. PMID- 11907760 TI - Re: Pulmonary steal from a left internal mammary artery bypass treated with transcatheter embolization. PMID- 11907761 TI - Re: Internal mammary artery pseudoaneurysm complicating central venous line placement: treatment with percutaneous thrombin injection. PMID- 11907768 TI - Interventional radiology in hemodialysis fistulae and grafts: a multidisciplinary approach. AB - PURPOSE: To review the place of interventional radiology in arteriovenous access for hemodialysis. METHODS: Prophylactic dilation of stenoses greater than 50% associated with clinical abnormalities such as flow-rate reduction is warranted to prolong access patency. Stents are placed only in selected cases with clearly insufficient results of dilation but they must never overlap major side veins and obviate future access creation. Thrombosed fistulae and grafts can be declotted by purely mechanical methods or in combination with a lytic drug. RESULTS: The success rates are over 90% for dilation, with frequent resort to stents in central veins. Long-term results in the largest series are better in forearm native fistulae compared with grafts (best 1-year primary patency: 51% versus 40%). The success rates for declotting are better in grafts compared with forearm fistulae but early rethrombosis is frequent in grafts so that primary patency rates can be better for native fistulae from the first month's follow-up (best 1 year primary patency: 49% versus 26%). CONCLUSION: Radiology achieves results comparable with surgery, with minimal invasiveness and better venous preservation. However, wide variations in the results suggest that the degree of commitment of physicians might be as important as the type of technique used. PMID- 11907769 TI - Embolization for hemoptysis: a six -year review. AB - PURPOSE: To review our method of embolization for hemoptysis. METHODS: Between 1993 and 1999, 134 patients were treated in our department for hemoptysis. One hundred and sixteen patients were followed up (18 were lost to follow-up) over a period ranging from 1 to 66 months (median 9.5 months, SD 14.81 months). Most cases were due to tuberculosis (83.6%) and malignancy (9.5%). One hundred and three required embolization. Vascular access was obtained via the femoral route but two cases required a brachial approach for abnormal branches of the subclavian artery. All abnormal vessels found were embolized using polyvinyl alcohol particles alone or in combination with gelfoam. RESULTS: Bronchial artery hypertrophy was found in 88.3% of cases; about a third of which had a nonbronchial systemic contribution. No angiographic abnormalities were found in 11.2%. Our failure rate was 18.4% (58% required surgery while 42% died from massive hemoptysis). Sixteen cases required multiple embolization sessions. No major complications were encountered. CONCLUSION: Embolization is effective for treatment of moderate to massive hemoptysis. The majority of our cases were due to tuberculosis. Approximately one third had nonbronchial systemic artery contributions, indicating that a concerted search for these is mandatory. PMID- 11907770 TI - Embolization of large aneurysms with long wire coils. AB - The authors report the experience of using long coils of 2 m length in the management of large aneurysms. Knowledge of the characteristics of these coils is of value for correct placement. These coils are safe and cost-effective for excluding large aneurysms. PMID- 11907771 TI - Role of interventional radiology in the treatment of biliary strictures following orthotopic liver transplantation. AB - PURPOSE: To evaluate the efficacy and safety of percutaneous treatment of biliary strictures complicating orthotopic liver transplantation (OLT). METHODS: Between October 1990 and May 2000, 619 patients underwent 678 liver transplants. Seventy of the 619 (11%) patients were found to be affected by biliary strictures by July 2000. Bilioplasty was performed in 51 of these 70 (73%) patients. A cohort of 33 of 51 (65%) patients were clinically followed for more than 12 months after the last percutaneous treatment and included in the survey results. RESULTS: After one to three treatments 24 of 33 (73%) patients were stricture-free on ultrasound and MR cholangiography follow-up. A delayed stricture recurrence required a fourth percutaneous bilioplasty in two of 33 (6%) patients. A surgical bilioenteric anastomosis was performed in six of 33 (18%) patients. Retransplantation was performed due to ischemic damage in one of 33 (3%) patients. CONCLUSION: Interventional radiology is an effective therapeutic alternative for the treatment of most biliary strictures complicating OLT. It has a high success rate and should be considered before surgical interventions. Elective surgery may be necessary in a few failed cases or those with more severe and extensive biliary strictures. PMID- 11907772 TI - Transcatheter thrombolysis with high-dose bolus tissue plasminogen activator in iatrogenic arterial occlusion after femoral arterial catheterization. AB - PURPOSE: To assess the efficacy of percutaneous local thrombolysis with high-dose bolus recombinant tissue plasminogen activator (rt-PA) in patients with acute limb ischemia due to arterial thrombosis after cardiac catheterization. METHODS: We treated eight patients (7 men; mean age 56 years) with thrombotic occlusion of both the common femoral artery (CFA) and external iliac artery (EIA) in six patients and of the CFA only in two patients. Two 5 mg boluses of rt-PA were injected into the proximal clot through a 5 Fr end-hole catheter and subsequently two additional boluses of 5 mg rt-PA were given through a catheter with multiple side-holes. In case of a significant amount of residual thrombus, a continuous infusion of 2.5 mg/hr of rt-PA was started. RESULTS: Successful lysis was achieved in all patients. The mean duration of lysis was 2 hr 41 min. The mean total amount of rt-PA delivered was 23.16 mg. In four patients unmasked flow limited dissections confined to the CFA were managed by prolonged balloon dilatation, while in the remaining four patients with extension of the dissection to the external iliac artery one or two Easy Wallstents were implanted. There was prompt relief of lower limb ischemic symptoms and signs in all patients. Two groin hematomas were conservatively treated. Clinical and color Doppler flow imaging follow-up with a mean duration of 15 months, showed no reappearance of ischemic symptoms or development of restenosis in any of the patients. One patient died 6 months after thrombolysis. CONCLUSIONS: Transcatheter thrombolysis with high-dose bolus rt-PA is a safe and effective treatment in patients with iatrogenic arterial occlusion after femoral catheterization. Underlying dissections should be treated by prolonged balloon dilatation but stent implantation is often required. PMID- 11907773 TI - Contrast-enhanced magnetic resonance angiography MoBI-trak in the study of peripheral vascular disease. AB - PURPOSE: To evaluate the diagnostic accuracy of contrast-enhanced magnetic resonance angiography (CE-MRA) MoBI-trak of peripheral vessels in patients with peripheral vascular disease who were candidates for digital subtraction angiography (DSA). METHODS: Eleven patients underwent CE-MRA with automated table movement (MoBI-trak) using a 1.5 T superconducting magnet (Philips Gyroscan ACS NT) equipped with a Power Trak 6000 gradient. Contrast medium (Gd-DTPA) was administered in two sequential boluses-20 cm(3) at 0.6 cm(3)/sec (starting phase) and 20 cm(3) at 0.3 cm(3)/sec (maintenance phase)-using a MedRad Spectris automatic injector. DSA was the gold standard and was performed using a Philips Integris 3000, with a brilliance intensifier of 38 seconds. DSA and MRA were evaluated on printed films. RESULTS: DSA provided 213 diagnostic assessments: 144 negative, 30 stenosis <50%, 5 stenoses in the 51-70% range, 12 stenoses in the 71 99% range and 22 occlusions. CE-MRA MoBI-trak sensitivity, specificity, positive and negative predictive value and diagnostic accuracy were 94.1%, 99.2%, 98.4%, 80.0% and 97.5%, respectively. CONCLUSIONS: Our work is in accordance with the literature. Using this technique MoBI-trak has been shown to be a reliable technique for the detection of peripheral vascular disease up to the trifurcation, although it underlines the necessity for more diagnostic investigation and improvements in the technique. PMID- 11907774 TI - Gadodiamide as an alternative contrast agent in intravenous digital subtraction angiography and interventional procedures of the upper extremity veins. AB - PURPOSE: Patients with renal insufficiency or other contraindications to iodine based contrast agents present a significant management dilemma when angiography or interventional procedures are required. We describe the use of gadolinium as an alternative contrast agent for intravenous digital subtraction angiography (IV DSA) and interventional procedures of the upper extremity veins. METHODS: Sixteen patients with symptoms of with moderate renal insufficiency underwent digital subtraction angiography for upper extremity and subclavian venography. Angiographic studies were obtained using gadolinium as a contrast agent. In six patients we used stents. RESULTS: Diagnostic images were obtained in all cases. No patient suffered a complication related to the use of gadolinium, and no patient demonstrated worsened renal function after the procedure. CONCLUSION: In the setting of a contraindication to iodine-based contrast agents, gadolinium represents an important alternative contrast material that allows for the visualization of and interventional procedures in the upper extremity veins. PMID- 11907775 TI - Vessel wall reaction after vena cava filter placement. AB - PURPOSE: To evaluate the interaction between the Cordis Keeper vena caval filter and vessel wall in a porcine model. METHODS: Implantation of the filter was performed in five pigs. Radiologic data concerning inferior vena cava (IVC) diameter and filter patency, filter leg span, and stability were collected. At 2 or 6 months post-implantation, histopathologic analysis of the IVC wall was performed. RESULTS: All filters remained patent with no evidence of migration. However, at 6 months follow-up, two legs of one filter penetrated the vessel wall and were adherent to the liver. These preliminary results suggest that with the observed gradual increase in the filter span, the risk of caval wall penetration increases with time, especially in a relatively small IVC (average diameter 16 mm). CONCLUSION: The Cordis Keeper filter was well tolerated, but seems to be prone to caval wall penetration in the long term. PMID- 11907776 TI - Meclofenamic acid for inhibition of human vascular smooth muscle cell proliferation and migration: an in vitro study. AB - PURPOSE: The aim of the study was to examine the effects of meclofenamic acid on proliferation, clonogenic activity, migratory ability, cell cycle distribution and p44/42 MAPK (mitogen activated protein kinase) expression in serum-stimulated human aortic smooth muscle cells (haSMCs). METHODS: haSMCs were treated with meclofenamic acid in three different concentrations (10 mM, 100 mM, 200 mM) for 4 days. Then meclofenamic acid-free culture medium was supplemented until day 20. Growth kinetics were assessed. Cell cycle analysis was performed by flow cytometry. Clonogenic activity was evaluated with colony formation assays. Migratory ability was investigated by stimulation with platelet-derived growth factor (PDGF-BB) in 24-well plates with 8 mm pores membrane inserts. p44/42 MAPK was detected by Western blot technique. RESULTS: Meclofenamic acid inhibited the proliferation, clonogenic activity and migratory ability of haSMCs in a dose dependent manner. Cell cycle analysis revealed a G2/M-phase block. The p44/42 MAPK was significantly reduced. CONCLUSION: Meclofenamic acid inhibits the proliferation and migration of haSMCs. If a sufficient dose of meclofenamic acid can be applied systemically or by local drug delivery it could be a valuable substance to prevent restenosis after angioplasty. PMID- 11907777 TI - Endovascular repair of a non-contained aortic rupture caused by a penetrating aortic ulcer. AB - This case report describes the endovascular treatment of an acute non-contained rupture of the descending aorta with a stent-graft as an emergency procedure. The aortic rupture was caused by a penetrating aortic ulcer. One year follow-up documents the complete recovery of the patient. PMID- 11907778 TI - Successful exclusion of a high internal carotid pseudoaneurysm using the Wallgraft endoprosthesis. AB - A 43-year-old woman presented with a several-month history of transient ischemic attacks 7 years following surgery for a malignant carotid body tumor. Angiography revealed a pseudoaneurysm at the distal vein graft anastomosis and a stenosis related to the proximal anastomosis. Due to the extensive previous surgery an endovascular approach was advocated and the pseudoaneurysm was successfully excluded using a covered stent (Wallgraft). This is, to our knowledge, the first time such an approach has been used following carotid body tumor excision. PMID- 11907779 TI - Basilar artery aneurysm treated with coil embolization via persistent primitive trigeminal artery. AB - A saccular aneurysm of the right basilar-superior cerebellar artery associated with a persistent primitive trigeminal artery (PPTA) was successfully treated by endovascular occlusion with Guglielmi detachable coils. Since both vertebral arteries and the basilar artery proximal to the junction with the PPTA were hypoplastic, a microcatheter was advanced via the PPTA. To our knowledge, this is the first case report describing the treatment of the aneurysm through the PPTA. PMID- 11907780 TI - Successful emergency stenting of acute ruptured false iliac aneurysm. AB - A 75-year-old man complaining of acute abdominal pain, 1 month after elective surgical repair of an abdominal aortic aneurysm by an aortabi-iliac bypass graft, was referred and admitted to the emergency room. Imaging by sonography and computed tomography scan revealed a ruptured iliac pseudoaneurysm at the right iliac anastomotic site with associated large retroperitoneal hematoma. We inserted a self-expanding covered Z-stent graft by a transfemoral approach and the iliac anastomotic pseudoaneurysm was successfully excluded. Our case demonstrates the possibilities of an endovascular approach for providing a fast, efficient and less aggravating procedure in order to treat these life-threatening conditions. PMID- 11907781 TI - Non-invasive investigation and management of aortic saddle embolus in a 7-month old infant. AB - There are few reports in the literature on the ultrasound appearance of aortic saddle embolus, and none relating to small children. This unusual condition is usually diagnosed angiographically. The purpose of this report is to show how effectively high-frequency ultrasound can identify a saddle embolus with its associated collateral circulation in a young child, and to demonstrate its usefulness in monitoring the efficacy of treatment. In this case the embolus occurred as a complication of parvovirus B19 myocarditis and was diagnosed and followed up entirely by ultrasound examination, with no invasive procedure performed. The early development of an extensive collateral circulation prevented distal tissue necrosis and allowed a conservative approach to management. PMID- 11907782 TI - Re: Two-stage transcatheter arterial embolization of a large hepatic artery pseudoaneurysm causing obstructive jaundice. PMID- 11907783 TI - Re: Rescue platelet glycoprotein IIb/IIIa inhibitor therapy in thrombosis complicating superficial femoral artery angioplasty. PMID- 11907785 TI - Leukemia- and lymphoma-associated genetic aberrations in healthy individuals. AB - In peripheral blood of at least 50% of healthy individuals, the translocations t(9;22) BCR/ABL, t(14;18) IgH/BCL-2, t(2;5) NPM-ALK and MLL duplications, which characterize chronic myelogenous leukemia and acute lymphoblastic leukemia, follicular lymphoma, anaplastic large cell lymphoma, and acute myelogenous leukemia, respectively, are detectable by sensitive polymerase chain reaction (PCR). No structural differences between these aberrations in normal or disturbed hematopoiesis are apparent. While the total count of t(9;22)- and t(14;18) positive cells does not exceed 10(4), those with MLL duplications are more frequent and account for approximately 10(7) cells in the total blood pool. t(14;18)-positive cells seem to be immortalized, but the biological consequences of the other aberrations in positive healthy persons have not been studied in detail. Due to the high frequency of positive individuals, most of them will not suffer from the correspondent leukemia or lymphoma, and criteria for subgroups that may be at a higher risk remain to be determined. Most likely, the number of genetic aberrations in healthy individuals, which so far are only associated with hematopoietic disorders, will increase in the near future. PMID- 11907786 TI - Thrombocytopenia associated with c7E3 Fab (abciximab). AB - Abciximab (c7E3 Fab) inhibits platelet aggregation and is used to prevent complications of percutaneous coronary intervention. Thrombocytopenia is an often cited complication of abciximab. Pseudothrombocytopenia is due to ethylenediaminetetraacetate (EDTA)-activated platelet agglutination, resulting in a spuriously low platelet count. We have looked at both "true" and pseudothrombocytopenia after infusion of abciximab. Sixty-six patients receiving their first exposure to abciximab after an unstable coronary event/revascularization were eligible. All the patients received a bolus of c7E3 Fab followed by a continuous infusion. Platelets were monitored in all patients at 2, 4, 12, 24, and 48 h, and more frequently if required. The incidence of thrombocytopenia and acute severe thrombocytopenia (platelet count < or =20,000/microl) was evaluated. A peripheral blood smear was performed on all patients showing thrombocytopenia to evaluate for pseudothrombocytopenia. Seventeen (25.6%) developed thrombocytopenia and nine (13.6%) developed acute severe thrombocytopenia. However, 18 of these patients had pseudothrombocytopenia. The onset of true thrombocytopenia was at 4 h after the infusion, while pseudothrombocytopenia occurred at anytime during the first 24 h. Only two (3.03%) patients required platelet transfusions. No life-threatening hemorrhagic complications were recognized. Five of six subjects with true thrombocytopenia had positive laboratory findings of disseminated intravascular coagulation; however, none had an adverse outcome. Acute severe thrombocytopenia was noted to be a relatively benign adverse effect of abciximab. There is an increasing incidence of pseudothrombocytopenia in this subgroup of patients. It would be worthwhile examining a peripheral blood smear or collecting blood for platelet counts in a heparin-coated tube in order to exclude this phenomenon and thereby prevent inappropriate discontinuation of this drug. PMID- 11907787 TI - Interleukin-1 beta, transforming growth factor beta 1, prostaglandin E2, and fibronectin levels in the conditioned mediums of bone marrow fibroblast cultures from lung and breast cancer patients. AB - We analyzed the ability of the bone marrow (BM) stromal cells to achieve confluence and their proliferative capacity in BM primary cultures from 30 untreated lung cancer patients (LCP), 27 breast cancer patients (BCP), and 30 normal controls (NC) when these confluent cells were induced to proliferate following four continuous subcultures. Moreover, we evaluated the production of interleukin-1 beta (IL-1beta), transforming growth factor beta 1 (TGF-beta1), fibronectin, and prostaglandin E2 (PGE2) by pure fibroblasts (fourth passage). A fibroblast colony-forming units (CFU-F) assay was used to investigate the proliferative and confluence capacity. Levels of IL-1beta, TGF-beta1, and fibronectin in conditioned mediums (CM) of fibroblast cultures were measured by enzyme-linked immunosorbent assay (ELISA) kit and PGE(2) by radioimmunoassay (RIA) kit. Confluence was achieved in the 60% of LCP and 78% of BCP primary cultures compared with 100% of NC, and only fibroblasts from seven LCP and six BCP cultures had the capacity to proliferate following four subcultures. Levels of IL-1beta were below 10 pg/ml in both patient groups, while NC had a mean value of 5882.57+/-221.61 pg/ml. Levels of TGF-beta1 in BCP were lower than NC values ( P<0.05). LCP and BCP had significantly decreased levels of fibronectin when compared to NC values ( P<0.05 and P<0.01, respectively). Levels of PGE2 in LCP were higher compared to NC ( P<0.01). In conclusion, BM fibroblasts from LCP and BCP presented a defective proliferative and confluence capacity, and this deficiency may be associated with the alteration of IL-1beta, TGF-beta1, fibronectin, and PGE2 production. PMID- 11907788 TI - CD34+ progenitor cells in idiopathic (primary) myelofibrosis: a comparative quantification between spleen and bone marrow tissue. AB - Previous studies on idiopathic (primary) myelofibrosis (IMF) are in keeping with the finding of an increased number of circulating CD34+ progenitor cells. However, dependent on the fibrosclerotic changes, little knowledge exists about quantitative relationships of this precursor cell population in spleen and bone marrow. The purpose of this study was to determine the number of CD34+ cells at these sites by applying immunohistochemistry and morphometry. In seven patients with IMF and a control group without hematopathology (14 patients), CD34+ progenitors were identified in the red pulp of the spleen and also in bone marrow trephine biopsies by the monoclonal antibody QBEND10. Histological analysis showed no differences regarding size or cytological details of these cells at both sites in patients with IMF as well as in the controls. On the other hand, in the IMF group there was an unequal distribution with an approximate twofold increase in precursor cells in the spleen compared to the bone marrow. This result is opposed to corresponding values of the control group. Considering the quantity of progenitor cells in the normal bone marrow, no significant increase was revealed in IMF, but a marked pooling (more than fourfold of the normal value) in the spleen. In conclusion, our findings are in keeping with a increased number of splenic CD34+ progenitor cells and therefore support the hypothesis of a pronounced filtration and trapping of this precursor cell population by the spleen in IMF. PMID- 11907789 TI - A comparative study on demographic, hematological, and cytogenetic findings and prognosis in acute myeloid leukemia with and without leukemia cutis. AB - We studied the incidence of leukemia cutis (LC) in 381 consecutive patients with acute myeloid leukemia (AML) in a single institution and compared the demographic, hematological, and cytogenetic findings in AML patients with and without LC. We also examined the response to intensive chemotherapy, overall survival, and duration of remission in this patient population with regard to the presence of LC. The prevalence of LC was 3.7% in clinically diagnosed patients and 2.9% in biopsy-proven cases, respectively. Patients with and without LC did not differ with regard to age, sex, white blood cell counts, hemoglobin, fibrinogen, and platelet counts at diagnosis, but lactate dehydrogenase (LDH) was significantly higher in patients with LC. Various karyotype abnormalities were found, but in patients with LC numerical abnormalities of chromosome 8 were significantly more common ( P<0.0001). Patients with LC did not differ from patients without LC with regard to remission rate, but there was a trend towards shorter remission duration in patients with LC. We conclude that patients with LC have some features different from patients without this symptom. The increased frequency of numerical aberrations of chromosome 8 in patients with LC was the most interesting observation of our study. The pathophysiological significance of this finding remains to be determined. PMID- 11907790 TI - Busulfan, cyclophosphamide, and etoposide as high-dose conditioning regimen in patients with malignant lymphoma. AB - We investigated the efficacy and toxicity of the combination of busulfan, cyclophosphamide, and etoposide (Bu/Cy/VP-16) as a preparative regimen prior to autologous hematopoietic stem cell transplantation (ASCT) in patients with Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL). Fifty-three patients with recurrent ( n=30), refractory ( n=20), or high-risk ( n=3) lymphoma were enrolled. The 10 patients with HD and 43 with NHL (median age: 46 years, range: 18-64) received busulfan (16 mg/kg), cyclophosphamide (120 mg/kg), and etoposide (30 or 45 mg/kg) followed by ASCT. A total of 50 patients (94%) were consolidated in complete ( n=25) or partial ( n=25) remission, whereas 3 patients had chemoresistant disease before Bu/Cy/VP-16. Thirty-five patients (66%) had received prior radiotherapy (RT) excluding total body irradiation (TBI) as part of the conditioning regimen. The main nonhematological toxicities (grade II-IV according to the Bearman score) in 52 evaluable patients were mucositis (79%) and hepatic toxicity (15%). Severe veno-occlusive disease (VOD) occurred in three patients (5.8%) including one treatment-related death caused by VOD. Overall, treatment-related mortality was 3.8%. After a median follow-up for surviving patients of 21 months (range: 6-118), 20 patients (38%) are in continuous complete remission, 8 patients (15%) are alive in relapse, and 25 patients (47%) died. Probabilities of relapse, event-free survival, and overall survival at 3 years were 63% [95% confidence interval (CI): 48-79%], 31% (95% CI: 17-46%), and 43% (95% CI: 27-59%), respectively. In conclusion, Bu/Cy/VP-16 is an effective and well-tolerated conditioning regimen in patients with HD and NHL. Both toxicity and outcome were not significantly different in patients treated with 30 mg/kg and 45 mg/kg etoposide, respectively. The observed long-term results are even comparable to those published for other established high-dose protocols, including TBI-based regimens. However, further investigations are necessary to evaluate the value of Bu/Cy/VP-16 as a high-dose protocol for malignant lymphoma, especially in patients who have already received extensive RT. PMID- 11907792 TI - Hodgkin's disease and hypothermia: case report and review of the literature. AB - We report the development of hypothermia following induction chemotherapy in a patient with stage IV Hodgkin's disease, fever, and hypotension. A link between hypothermia, hypotension, and sensory neuropathy is possible. Acute autonomic neuropathy is the suggested cause. PMID- 11907791 TI - Occurrence of sarcoidosis subsequent to chemotherapy for non-Hodgkin's lymphoma: report of two cases. AB - Sarcoidosis-lymphoma syndrome is a well-established syndrome where sarcoidosis is followed by the development of a lymphoproliferative disease such as non Hodgkin's lymphoma (NHL). Here we report two patients with NHL who developed sarcoidosis subsequent to the diagnosis of lymphoproliferative disease. In both cases, chemotherapeutic treatment had already been initiated or was completed when sarcoidosis occurred. In these patients, sarcoidosis may have been triggered by immunologic aberrations induced by antineoplastic therapy or as a consequence of an underlying immunologic disturbance associated with the lymphoma. When a suspected relapse of lymphoma presents with signs and symptoms compatible with sarcoidosis, this rare immunologic disorder has to be ruled out by careful clinical and histopathologic analysis to prevent mistreatment. PMID- 11907793 TI - Isolated cavernous sinus extramedullary relapse of chronic myelogenous leukemia following allogeneic stem cell transplant. AB - Isolated extramedullary relapse of chronic myelogenous leukemia (CML) after allogeneic stem cell transplant is rare. A case is reported of a patient who developed a granulocytic sarcoma of the cavernous sinus 7 months after allogeneic transplant for CML. He presented with neurologic deficits, and a mass lesion was found on imaging. No evidence of hematologic relapse was identified by bone marrow histology or cytogenetics. A premortem diagnosis was not possible, and the patient died 2 months later of an intracerebral hemorrhage after receiving various therapies directed against a presumed infectious etiology. Granulocytic sarcoma should be considered in the differential diagnosis of mass lesions presenting after allogeneic transplant for CML, even if there is no evidence of medullary disease. PMID- 11907794 TI - Sweet's syndrome associated with retinoic acid syndrome in a patient with promyelocytic leukemia. AB - We report a case of Sweet's syndrome associated with retinoic acid syndrome in a patient with acute promyelocytic leukemia treated with all- trans retinoic acid (ATRA). Sweet's syndrome appeared on day 6 of ATRA therapy for promyelocytic leukemia. It was associated with a mild retinoic acid syndrome, an inflammatory syndrome occurring in 25% of patients treated with ATRA and characterized by features of capillary leakage with systemic inflammatory signs. The ATRA therapy was discontinued for 11 days and treatment with corticosteroids improved the systemic and cutaneous signs. Only 11 cases of Sweet's syndrome associated with ATRA have been previously reported in the literature, involving only the skin in eight cases, the skin and muscles in two cases, and the lung, kidney, fascia, and muscles in one case. Sweet's syndrome was followed by retinoic acid syndrome in one of these cases. The previously reported cases are reviewed, and the mechanisms of Sweet's and retinoic acid syndromes and the link between them are discussed. PMID- 11907795 TI - Testicular infiltration in acute myeloid leukemia with complex karyotype including t(8;21). AB - Testicular infiltration is a well-known complication in acute lymphoblastic leukemia. In acute myeloid leukemia (AML), it has rarely been described and preferably occurred in cases with myelomonocytic or monoblastic differentiation. We report on a patient with AML with complex karyotype including translocation t(8;21) who presented with testicular infiltration at the time of his third bone marrow relapse. Cytological analysis of the specimen showed infiltration with blasts displaying the typical morphology of AML with translocation t(8;21) and comparable to those detected in the bone marrow. Fine needle aspiration cytology might suffice in these cases and should be performed if testicular involvement is suspected. PMID- 11907796 TI - Anti-CD20 antibody as consolidation therapy in a patient with primary plasma cell leukemia after high-dose therapy and autologous stem cell transplantation. AB - In multiple myeloma (MM), circulating malignant B cells are proposed as the proliferative compartment of the disease. In view of the close relationship between multiple myeloma and primary plasma cell leukemia (PCL), an anti-CD20 antibody treatment might also be considered as consolidation for patients with PCL. A 55-year-old patient diagnosed with PCL achieved complete remission after autologous transplantation. A total of four weekly courses of rituximab (375 mg/m(2)) were administered. Prior to antibody therapy, CD20+ cells comprised 22.6% of the mononuclear cells in peripheral blood (PB) assessed by flow cytometry and were enriched by magnetic activated cell sorting (MACS). In the enriched CD20+ fraction, 0.093% clonotypic cells were detected using a quantitative polymerase chain reaction (PCR) assay based on limiting dilutions. The proportion of clonotypic cells was 0.034% in PB and 0.032% in bone marrow (BM). Rituximab depleted CD20+ cells completely in PB and BM. Tumor load in PB and BM at day 40 and in PB at day 70 did not change in comparison to prior to therapy (0.037% in PB, 0.026% in BM). At day 90, the tumor load increased to 0.066% in PB. At day 120, the patient relapsed with 0.65% CD38++/CD138+/CD20- plasma cells and furthermore no CD20+ B cells in PB. The expansion of plasma cells was accompanied by an increase in the tumor load in both compartments (PB: 0.65%, BM: 1.8%). The accumulation of plasma cells during disease progression without the reappearance of CD20+ cells did not sustain the role of circulating clonotypic B cells as proliferative compartment in our patient. However, it cannot be excluded that rituximab was not able to eradicate malignant B cells, which subsequently contributed to relapse. PMID- 11907799 TI - Immunocytochemical localization of synphilin-1, an alpha-synuclein-associated protein, in neurodegenerative disorders. AB - Alpha-synuclein is a major component of Lewy bodies (LB) in Parkinson's disease (PD) and dementia with LB (DLB), as well as of glial cytoplasmic inclusions (GCI) in multiple system atrophy (MSA). Recently, a novel protein called synphilin-1 has been identified that associates with alpha-synuclein, and it has been reported that co-transfection of both alpha-synuclein and synphilin-1 in mammalian cells yielded eosinophilic cytoplasmic inclusions resembling LB. Immunocytochemical and ultrastructural investigations have now been performed on the brain of patients with various neurodegenerative disorders using anti synphilin-1 antibodies. These antibodies immunostained the neuropil in a punctate pattern throughout the brain of control subjects. In PD, most LB observed in the brain stem were positive for synphilin-1. These LB showed intense staining in their central cores, but their peripheral portions were only weakly stained or unstained. Pale bodies and Lewy neurites, which were positive for alpha synuclein, were synphilin-1 negative. In DLB, a small fraction of cortical LB were immunolabeled by anti-synphilin-1. In MSA, numerous GCI were positive for synphilin-1. Immunoelectron microscopy revealed that the reaction product was localized within filamentous and circular structures in LB. Various neuronal and glial inclusions in neurodegenerative disorders other than LB disease and MSA were synphilin-1 negative. These findings suggest that abnormal accumulation of synphilin-1 is specific for brain lesions in which alpha-synuclein is a major component. PMID- 11907798 TI - Increased clusterin (apolipoprotein J) expression in human and mouse brains infected with transmissible spongiform encephalopathies. AB - Clusterin (apolipoprotein J), a multifunctional protein involved in amyloidogenesis in Alzheimer's disease, was studied immunohistochemically in both human transmissible spongiform encephalopathies (TSEs) and a mouse model of human TSE. Clusterin immunoreactivity was co-localized with plaque-type deposits but not with punctate-type prion protein (PrP) deposits in human TSEs. On the other hand, clusterin-positive astrocytes were readily demonstrated in the regions of punctate PrP deposits, but not around plaque PrP deposits despite the presence of surrounding astrocytes. Clusterin expression in astrocytes was not disease specific, but the punctate immunoreactivity for clusterin was more prominently demonstrated in TSEs with punctate PrP deposits. Serial analysis in the mouse model of human TSE revealed that clusterin expression in astrocytes was enhanced in the lesions with punctate-type PrP deposits during the disease progression. Thus, the induction of clusterin expression in astrocytes could be more enhanced by punctate-type PrP deposits than by plaque-type deposits. The clusterin molecules co-localized in plaque PrP deposits might be derived not from surrounding astrocytes but from other resources such as cerebrospinal fluid and blood plasma, both of which contain clusterin in significant amounts. Taken together with previously reported findings of the anti-amyloidogenic property in clusterin, our findings suggest that clusterin may be induced as one of the important molecules participating in the neurodegeneration caused by abnormally deposited PrP. PMID- 11907800 TI - Antioxidant agents have a different expression pattern in muscle fibers of patients with mitochondrial diseases. AB - Respiratory chain dysfunction leads to reactive oxygen species (ROS) generation with following oxidative stress and cellular damage. A histochemical and immunohistochemical study was performed on muscle biopsies from 17 patients with mitochondrial disease [chronic progressive external ophthalmoplegia (CPEO), mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), myoclonic epilepsy with ragged red fibers (MERRF)] to evaluate the expression pattern and location of manganese superoxide dismutase (MnSOD), copper zinc superoxide dismutase (CuZnSOD) and reduced glutathione (GSH) in skeletal muscle fibers. Our data showed that: (1) MnSOD, CuZnSOD and GSH are expressed in fibers with respiratory chain deficiency; (2) the antioxidant induction is correlated with the degree of mitochondrial proliferation, but not with clinical phenotype, patients' age, duration of disease, biochemical defects or mitochondrial DNA abnormalities. In addition, we suggest that expression of MnSOD and GSH may be considered an initial, indirect sign of respiratory chain dysfunction because it is observed in the early stages of the disease. PMID- 11907801 TI - Evidence for pathological involvement of the spinal cord in motor neuron disease inclusion dementia. AB - Fronto-temporal dementia (FTD), characterised pathologically by ubiquitinated inclusions in the hippocampal dentate fascia and fronto-temporal cortex, may develop in association with motor neuron disease (MND). However, FTD with identical pathological hallmarks may occur in isolation and has been termed motor neuron disease-inclusion dementia (MND-ID). We studied the pathology of three cases of MND-ID including the spinal cord, which has not previously been examined in this condition. The ages of the patients at death were 53, 70 and 68 years and the onset of FTD 10, 15 and 9 years before death. Neuropathological findings in all cases included micro-vacuolation in cortical layer II and ubiquitinated intraneuronal inclusions in fronto-temporal cortex and hippocampal dentate fascia. One case showed unusual basophilic, ubiquitinated neuronal cytoplasmic inclusions in the brain stem. Qualitative assessment of the spinal cord was normal in two cases, while the third showed mild pallor of the lateral cortico spinal tracts and ubiquitinated inclusions in motor neurons typical of MND. Morphometry did not reveal any significant loss or decrease in size of anterior horn motor neurons. The results of this study are consistent with the hypothesis that the pathological changes of pure MND and MND-ID form a spectrum and demonstrate that pathological involvement of the motor system may occur in MND-ID without clinical evidence of MND. PMID- 11907803 TI - Differential passage of [14C]sucrose and [3H]inulin across rat blood-brain barrier after cerebral ischemia. AB - The biophysical nature of blood-brain barrier (BBB) opening after ischemic or hemorrhagic stroke or traumatic brain injury is unresolved. Ultrastructural (electron micrograph) investigations of experimental BBB injury commonly indicate the abnormal presence of vesicles or tubular structures in cerebrovascular endothelial cells, suggesting the likelihood of convective, fluid-phase transport of blood substances into brain. We measured transfer constants (K(i)s) for the simultaneous passage of two intravenously delivered tracers ([14C]sucrose, mol wt=342; [3H]inulin approximately 5,000) across the intact BBB in the rat, and 24 h after global cerebral ischemia (16-20 min duration) or 24, 48 or 72 h after focal ischemia (2 h duration). In both ischemia models, the upward increment in K(i) (DeltaK(i)) for sucrose, indicating the extra injury-related tracer flux into brain, significantly exceeded that for inulin, as might be expected with faster diffusion of the smaller molecule through injury pores or channels. This inequality of DeltaK(i)s did not suggest a major role for convective, fluid-phase transport by endothelial vesicular or tubular structures and a predominance of diffusional transport was indicated. PMID- 11907802 TI - Three-dimensional and fractal analyses of assemblies of amyloid beta protein subtypes [Abeta40 and Abeta42(43)] in canine senile plaques. AB - The three-dimensional (3D) distribution of amyloid beta protein (Abeta) subtypes [Abeta40 and Abeta42(43)] in canine senile plaques (SP) was observed using a confocal laser scanning microscope. In diffuse plaques (DP), Abeta42(43) alone was deposited as an uneven nebula-like assembly of fine granules. The border of the Abeta42(43) assembly was unclear and diffusely merged to the surrounding area. Mature plaques (MP), on the other hand, showed two patterns of Abeta deposition. In some MP, only Abeta40 was deposited as a defined assembly of very short fibrillary structures. Other MP consisted of both Abeta40 and Abeta42(43), and the deposition patterns of the two Abeta species were the same as those in single-positive plaques; fine granular with unclear margin for Abeta42(43), and short fibrillary structures for Abeta40. Additionally, we calculated the fractal dimensions (FD) of both Abeta40 and Abeta42(43) assemblies, and examined the serial change of FD in each SP. The FD of Abeta42(43)-positive DP ranged from 1.05 to 1.27, and those of Abeta40-positive MP ranged from 1.13 to 1.54 in single positive plaques. In one double-positive MP, FD ranged from 1.02 to 1.36 for Abeta42(43) and from 1.01 to 1.51 for Abeta40. These results showed that the FD of canine Abeta40 assemblies was higher than that of Abeta42(43) assemblies, and the spatial changes of FD values for Abeta40 and Abeta42(43) in double-positive plaques were quite different. These morphological analyses clearly showed that canine DP and MP have completely different 3D structures, suggesting that their processes of formation are different. PMID- 11907804 TI - The Golgi apparatus is fragmented in spinal cord motor neurons of amyotrophic lateral sclerosis with basophilic inclusions. AB - The mechanisms of neuronal death in amyotrophic lateral sclerosis (ALS) are not known. A pathological aggregation of cytoplasmic constituents in the form of variety of inclusions may play a role in the pathogenesis of neuronal death. Cytoplasmic basophilic inclusions (BIs) in motor neurons are commonly found in sporadic juvenile ALS. The functional significance of these inclusions is not known, i.e., whether they represent a protective reaction for the isolation of abnormal products from the cytoplasm, or a sign of irreversible neuronal damage. To gain insights on the significance of BIs we asked whether neurons with BIs had an intact or fragmented Golgi apparatus (GA), a sign of neuronal degeneration reported not only in sporadic and familial ALS with mutations of the Cu/Zn superoxide dismutase gene (SOD1), but also in transgenic mice expressing the G93A mutation of SOD1. In these mice fragmentation of the GA of spinal cord motor neurons was found months before the onset of paralysis. We report here that all neurons bearing the inclusions showed fragmentation and reduced number of GA. These results suggest that common pathogenetic mechanisms are involved in the production of BIs and in the fragmentation of the GA. PMID- 11907805 TI - Embryonic hydromyelia: cystic dilatation of the lumbosacral neural tube in human embryos. AB - In a large collection of human embryos (the Kyoto Collection of Human Embryos, Kyoto University), we encountered five cases with abnormal dilatation of the neural tube at the lumbosacral level. In these examples, the central canal was enlarged, and the roof plate of the neural tube was extremely thin and expanded. The mesenchymal tissue was scarce or lacking between the roof plate and the surface ectoderm. This type of anomaly was assumed to be formed after neural tube closure and may be an early form of spina bifida. In two of the cases, some abnormal cells were found ectopically between the thin roof plate and the surface ectoderm. Morphologically, these cells resembled those forming spinal ganglia and could be of the neural crest origin. Since neural crest cells are pluripotent and can differentiate into a variety of tissues, such ectopic cells might undergo abnormal differentiation into teratomatous tumors and/or lipomas, which are frequently associated with spina bifida. We also discuss the definition of spina bifida and the classification of neural tube defects from the embryological and pathogenic viewpoints and propose a new classification of neural tube defects. PMID- 11907806 TI - Physical and functional characterization of the human LGI1 gene and its possible role in glioma development. AB - The human gene termed LGI1 (leucine-rich gene - glioma inactivated) has been isolated recently, and is supposed to be an additional candidate tumor suppressor gene involved in the formation and progression of glioblastoma multiforme [Chernova et al. (1998) Oncogene 17:2873-2881]. To test this hypothesis and to complete the characterization of the gene, we performed various detailed studies on the genomic structure, the mRNA expression level, the integrity of the cDNA, and retroviral gene transfer into LGI1-deficient cell lines. Two single nucleotide polymorphisms in the promotor region and a highly polymorphic intragenic microsatellite repeat between exon 4 and 5 were found. Phylogenetic sequence analysis techniques were applied, which showed functional relationships between LGI1 and TRK and SLIT protein families that are known to be involved in development and maintenance of the nervous system. Fluorescence in situ hybridization (FISH) analysis showed LGI1 to be present on 10q24 in each of 11 glioma-derived cell lines evaluated. Sequence analysis of the LGI1 transcript did not detect any mutation. Relative amounts of LGI1 mRNA copy numbers as measured by the real-time fluorescence detection LightCycler technology differed more than three orders of magnitude and were significantly reduced in 10 of 11 cell lines. Retroviral gene transfer into LGI1-deficient glioma-derived cell lines could not substantiate any difference to control infected cultures regarding growth rate, S phase transition, and maintenance of marker gene expression. The strong homology to proteins involved in development, differentiation, or maintenance of the nervous system provides evidence for a function of the LGI1 protein in neural tissue. The observation that translocation or deletion of the LGI1 locus or mutation of the coding sequence of the LGI1 mRNA is not a frequent event in malignant glioma cell lines suggests that epigenetic factors lead to substantial differences in the amount of LGI1 mRNA expression. In addition, that the effect is lacking after retroviral gene transfer in cell culture suggests that binding of some kind of a ligand is essential for its biological activity. PMID- 11907807 TI - Phenotype versus genotype correlation in oligodendrogliomas and low-grade diffuse astrocytomas. AB - Oligodendrogliomas typically show loss of heterozygosity (LOH) on chromosomes 1p and 19q, which correlates with their response to chemotherapy, whereas low-grade astrocytomas are characterized by frequent TP53 mutations and lack of sensitivity to alkylating therapeutic agents. Unequivocal histological distinction of low grade diffuse astrocytomas from oligodendrogliomas and oligoastrocytomas is often difficult. To elucidate the relationships between morphological phenotype and genetic profile, we screened 19 oligodendrogliomas (WHO grade II) and 23 low grade diffuse astrocytomas (WHO grade II) for TP53 mutations and LOH on 1p and 19q. In oligodendrogliomas, LOH on chromosomes 1p and/or 19q was found in 15 cases (79%) and TP53 mutation was detected in 4 cases (21%). The presence of a typical perinuclear halo in >50% of tumour cells and a chicken-wire vascular pattern were significantly associated with LOH on 1p or 19q (93% of cases). This suggests that oligodendrogliomas with classical histologic features are likely to have a better prognosis. In low-grade diffuse astrocytomas, LOH on chromosomes 1p and/or 19q was found in three cases (13%) and TP53 mutation was detected in ten cases (43%). Histologically, five low-grade astrocytomas (22%) contained small areas with oligodendroglial differentiation, but this did not correlate with the presence of TP53 mutations or LOH on 1p and 19q. In both oligodendrogliomas and astrocytomas, LOH on chromosomes 1p or 19q and TP53 mutation were mutually exclusive. Methylation of the promoter of the gene for O (6)-methylguanine-DNA methyltransferase (MGMT), a DNA repair protein, which confers resistance to chemotherapy with alkylating agents, was detected in 47% of oligodendrogliomas and 48% of low-grade diffuse astrocytomas. There was no correlation with LOH on chromosomes 1p/19q, suggesting that MGMT may not be a prognostic marker for oligodendrogliomas. PMID- 11907808 TI - Pallido-nigral spheroids in nonhuman primates: accumulation of heat shock proteins in astroglial processes. AB - Alpha-B crystallin, ubiquitin and heat shock protein 27 (hsp27) belong to a class of proteins that are overexpressed in response to pathological conditions associated with increased cellular stress. In the present study, brain sections of old rhesus monkeys ( Macaca mulatta; n=10; mean age, 29.4 years) and baboons ( Papio anubis; n=8; mean age, 18.3 years) were examined for ubiquitin, alpha-B crystallin and hsp27-immunopositive structures. In both species, immunoreactive spheroid-like bodies were found in the globus pallidus and in the substantia nigra, pars reticulata. These structures frequently were associated with abnormally swollen cellular processes. To further clarify the origin of the pallido-nigral spheroids, single- and double-immunostaining was performed for hsp27, alpha-B crystallin and the astroglial marker glial fibrillary acidic protein (GFAP) as well as for neuronal markers against neurofilament and dendritic microtubule-associated protein 2. Confocal microscopic analysis demonstrated that spheroids were localized in swollen astroglial processes, whereas they were not seen in neuronal structures. Thus, pallido-nigral spheroids can be classified as astroglial accumulations of heat shock proteins. Further investigations of these structures may provide information pertinent to our understanding of astroglial heat shock protein inclusions developing in degenerative human brain diseases. PMID- 11907809 TI - Neuropathology of Raine syndrome. AB - We present three cases of Raine syndrome occurring in siblings of consanguineous parents. Raine syndrome is characterised by generalised osteosclerosis with craniofacial anomalies and intracranial calcifications. So far, only nine cases have been reported, and no evaluation of the distribution and extent of the cerebral mineralisations, as well as their impact on the surrounding tissue, has been undertaken yet. In our cases, calcifications were unevenly distributed throughout the central nervous system, not associated with neuronal loss or dystrophic events and appeared mostly as single calcospherites within the neuropil with occasional confluent deposits at advanced gestational age. There was intense perifocal microgliosis around single immature calcospherites, as well as mild astrogliosis around and within the confluent lesions, in which occasional macrophages could be found. Rarely, mineralisations occurred in blood-vessel walls, mainly affecting basal ganglia. Preferential sites of calcification were parietal and occipital periventricular white matter and corpus callosum, while frontal lobes were mildly affected. The cortex, temporal lobes as well as internal capsule, brain stem, cerebellum, leptomeninges, pituitary gland and choroid plexus were devoid of mineralisations. The subcortical grey matter was moderately involved in the putamen and pallidum, mildly in the caudate nucleus and subependymal germ cell matrix and not at all in the thalamus, Ammon's horn, amygdala and substantia nigra. The distribution of mineral deposits was thus inversely correlated to regional blood circulation and capillary density, with calcifications being concentrated in more sparsely perfused areas but lacking in highly vascularised tissue. This inverse relationship between mineralisation and regional blood flow was reflected in the varying distribution of calcospherites in grey and white matter as well as in the white matter of different lobes. PMID- 11907810 TI - A case of clinically and neuropathologically atypical corticobasal degeneration with widespread iron deposition. AB - A 65-year-old woman was admitted to our hospital for forgetfulness, depression and eccentric behavior that had been first noticed 2 years prior to admission. She showed memory impairment, perseveration and repeated violent actions, but no limb-kinetic apraxia. She died 12 years after the onset of symptoms. At autopsy, the unfixed brain weighed 820 g. Atrophy was circumscribed in the frontal lobe on both sides. The globus pallidus and the caudate nucleus were markedly atrophic and gold yellow in color, and the substantia nigra was strikingly pale. The cortical area showed neuronal loss and status spongiosus of the second and third cortical layers with ballooned neurons. Marked neuronal loss was observed in the dorsomedial nucleus of the thalamus, Meynert basal nucleus and substantia nigra. With Holzer stain, fibrillary gliosis was found to be severe in the frontal lobe, globus pallidus, subthalamic nucleus, hippocampus, dorsomedial nucleus of thalamus, substantia nigra, pontine tegmentum and inferior olivary nucleus. With Bielschowsky-Hirano stain, neurofibrillary tangles were observed in the cortex, hippocampus, substantia nigra, dentate nucleus, subthalamic nucleus, pontine nucleus, the inferior olivary nucleus, dorsomedial nucleus of the thalamus and, to a lesser extent, the neostriatum. Strikingly numerous argyrophilic and tau positive threads were present in the cerebral white matter. These neuropathological findings corresponded to corticobasal degeneration, but lesions characteristic of progressive supranuclear palsy were also found. Moreover, widespread iron deposition throughout the central nervous system was the most striking finding of the present case. To our knowledge, such a case has not been reported in the literature to date. PMID- 11907811 TI - A case of rapidly progressive multiple system degeneration: morphological findings and pathogenetic implications. AB - A 62-year-old woman was referred to our psychiatric hospital by the municipal health office, because she was in such a neglected condition that she was a danger to herself. Initially, it was suspected that she was suffering from dementia or psychosis. X-rays led to the suspicion of bronchial carcinoma. Consequently, the mental changes were interpreted as probable effects of metastases to the brain. There was not enough time, however, to check the patient thoroughly to find the actual cause of her altered personality. The patient developed a high fever (up to 42 degrees C). A few days later, she died of cardiorespiratory failure with severe abscess-forming obstructive pneumonia. An autopsy confirmed the bronchial carcinoma. However, metastases were only found in the hilar lymph nodes. No metastases were detectable in the CNS, either macroscopically or microscopically. The neuropathological examination of the brain revealed multiple system degeneration. The striking microscopic findings (a large number of typical apoptotic figures visible with the light microscope in ganglion cells, lack of cytoplasmic inclusion bodies in the oligodendroglia and an unusually strong monocytic reaction (so-called reactive satellitosis) indicated that the disease course had been very rapid. Perhaps these were early steps in the evolution of a multisystem atrophy. A kind of time-lapse effect, as it were, revealed simultaneously individual details of the pathogenetic course, which would have disappeared in the usual long course of the disease and thus could not have been observed, even with the most up-to-date molecular methods. It is very likely that this was a paraneoplastic syndrome. PMID- 11907812 TI - Rhombencephalosynapsis with massive hydrocephalus: case report and pathogenetic considerations. AB - Rhombencephalosynapsis (RS) is a rare cerebellar malformation characterized by vermian agenesis or hypogenesis, fusion of the hemispheres, and closely apposed or fused dentate nuclei. This report concerns a 29-year-old Caucasian female with a history of profound retardation and chronic hydrocephalus. Multiple shunts had been placed and there had been several admissions for shunt infections. Autopsy confirmed massive hydrocephalus and revealed marked aqueductal stenosis. The cerebellum was small with vermian aplasia, closely apposed dentate nuclei, fusion of the middle cerebellar peduncles, and a small fourth ventricle. Associated defects included fusion of the inferior colliculi and absence of dorsal olivary nuclei. This spectrum of anomalies is consistent with RS and could be explained by an embryological defect of dorsal pattering that affects the "isthmic organizer" at the mesencephalic-metencephalic border. Molecular analysis of dorsalizing genes, such as Lmx1a, which regulate early developmental events at the pontomesencephalic junction, may reveal a mutation or mutations unique to RS. PMID- 11907813 TI - Rhombencephalosynapsis. PMID- 11907814 TI - Microdysgenesis--relevant finding in refractory epilepsy? A response to S. H. Eriksson et al., Acta Neuropathologica (2002) 103:74-77. PMID- 11907816 TI - Oxygen sensing, HIF-1alpha stabilization and potential therapeutic strategies. PMID- 11907818 TI - Dofetilide block involves interactions with open and inactivated states of HERG channels. AB - Rapidly activating delayed rectifier current ( IKr) is the key target of class III antiarrhythmic drugs including dofetilide. Due to its complex gating properties, the precise channel state or states that interact with these agents remain poorly defined. We have undertaken a careful analysis of the state dependence of HERG channel block by dofetilide in Xenopus oocytes and Chinese Hamster Ovary (CHO) cells by devising a protocol in which brief sampling pulses were superimposed over a wide range of test potentials. The rate of block onset, maximal steady-state block and IC50 were similar for all test potentials over the activation range, demonstrating that the drug probably interacts with open and/or inactivated but not resting HERG channels with high affinity. Reducing the fraction of inactivated channels at 0 mV by augmenting the external potassium concentration did not alter the sensitivity to dofetilide. In contrast, the S631A and S620T HERG mutations both eliminated inward rectification and reduced dofetilide affinity by approximately 10- and approximately 100-fold respectively. We have also found a novel ultra-slow activation process which occurs in wild type HERG channels at threshold potentials. Overall, our data imply that dofetilide block occurs equally at all voltages positive to the activation threshold, and that the drug interacts with HERG channels in both the open and inactivated states. PMID- 11907817 TI - Characterization of the calcium release domains during excitation-contraction coupling in skeletal muscle fibres. AB - The spatiotemporal properties of the Ca2+ release process in skeletal muscle fibres were determined using a confocal spot detection system. The low-affinity, fluorescent Ca2+ indicator Oregon Green 488 BAPTA-5N (OGB-5N) was used to record localized, action potential-induced fluorescence signals from consecutive locations separated by 200 nm within a single sarcomere. Three-dimensional reconstructions of the Ca2+ transients illustrated the existence of fluorescence domains around Ca2+ release sites, which are centred at the T-tubules. By constructing isochronal plots, it was estimated that the earliest detectable full width at half-maximum (FWHM) of the Ca2+ domains was 0.77+/-0.08 microm and increased rapidly with time to 1.4+/-0.04 microm at peak (17-18 degrees C). A delay of 0.64+/-0.1 ms was observed between the onset of the fluorescence transients at the Z- and M-lines. Deconvolution of fluorescence transients gave estimates of approximately 9 and 2 microM for the peak [Ca2+] changes at the Z and M-lines, respectively. Our results are compatible with the possibility that action potential stimulation elicits Ca2+ release from a region of the sarcoplasmic reticulum (SR) broader than the T-SR junction. PMID- 11907819 TI - Multiple inhibitory effects of zatebradine (UL-FS 49) on the electrophysiological properties of retinal rod photoreceptors. AB - Effects of the bradycardiac agent zatebradine (UL-FS 49) were determined in intracellular and patch-clamp experiments of amphibian rod photoreceptors. Zatebradine (0.3-100 microM) greatly enhanced the bright-light-induced hyperpolarization of membrane potential in frog retinal rods and caused damped oscillations during the membrane potential recovery phase. In inner segments of the rod photoreceptor cells, the hyperpolarization-activated inward current ( Ih) contributes to the recovery of the photoresponse. Patch-clamp recordings from newt rod photoreceptor cells revealed that zatebradine caused use- and concentration-dependent (0.1-100 microM) inhibition of Ih: conductance was reduced without effects on the reversal potential or activation voltage. Our data confirmed that the pharmacological properties of Ih in rod photoreceptors were similar to those of Ih in cardiac myocytes. In addition, zatebradine inhibited voltage-gated outward K+ currents ( IK), but did not affect L-type Ca2+ currents ( ICa). These results are consistent with the inhibition of IK and Ih by zatebradine in other cells, and may explain the oscillations evoked during the recovery phase of the membrane potential. These multiple actions of zatebradine on channels in rod photoreceptors may explain its effects on the electroretinogram (ERG) in vivo and its adverse effects on vision in clinical studies. PMID- 11907820 TI - Recombinant Kv1.3 potassium channels stabilize tonic firing of cultured rat hippocampal neurons. AB - We transfected cultured hippocampal neurons with the cDNA of the voltage-gated K+ channel Kv1.3 to investigate the mechanisms by which a specific ion channel influences excitability. In transfected neurons under voltage clamp we observed an additional outward current that was blocked selectively by margatoxin. Under current-clamp conditions, Kv1.3-expressing neurons fired tonically over a wide range of stimulation intensity. In non-transfected neurons, or in Kv1.3 expressing cells blocked with margatoxin, only a few action potentials were elicited before a stationary depolarized state was reached. We attribute the specific effect of Kv1.3 to its particularly slow deactivation near the resting potential. A computational model showed that a continuous outwards current arises in Kv1.3-expressing neurons during the interspike intervals. It expands the dynamic range so that these neurons still fire tonically at stimulus current intensities at which non-transfected cells have already been driven into a stationary depolarized state. PMID- 11907821 TI - Postural responses of forelimb extensors to somatosensory signals elicited during wrist rotation: interaction with vestibular reflexes. AB - The responses of the forelimb extensor triceps brachii (TB) to sinusoidal wrist rotation, at 0.156 Hz, +/-5 degrees, were investigated in decerebrate cats. In most of the tested muscles [12/15) the multiunit electromyographic (EMG) activity of the TB muscle increased during dorsiflexion and decreased during plantar flexion of the ipsilateral limb extremity, showing an average (+/-SD) gain of 1.38+/-0.64 impulses/s/ degrees and a phase lead of 33+/-19 degrees with respect to the extreme dorsiflexion of the forepaw. Both parameters remained unmodified by increasing the amplitude of stimulation from 5 degrees up to 10 degrees. Rotation of the contralateral wrist had no effect on TB activity. When the rotation of the ipsilateral wrist occurred during sinusoidal roll tilt of the whole animal, a stimulus that activates vestibular receptors, the TB response to combined stimulation closely corresponded to the vectorial sum of the individual responses. Dorsiflexion of the forepaw could drive the vestibulospinal system, which excites the ipsilateral TB motoneurons. In fact, functional inactivation of the cerebellar anterior vermis, which controls vestibulospinal neurons, significantly modified the TB response to wrist rotation. It appears, therefore, that the somatosensory signals elicited by wrist rotation utilize the spinocerebellum to modulate the activity of the TB. The TB responses to wrist rotation could play a role in stabilizing posture during stance and locomotion. PMID- 11907822 TI - The outwardly rectifying chloride channel in rat peritoneal mast cells is regulated by serine/threonine kinases and phosphatases. AB - A slowly activating, outwardly rectifying Cl- channel (ORCC) has been described in rat peritoneal mast cells (RPMCs). This channel is activated by intracellular application of cAMP, an effect that might be mediated by a PKA-type serine/threonine protein kinase. To test this hypothesis, whole-cell patch-clamp experiments (nystatin-perforated patch) were performed and 8-bromoadenosine 3',5' cyclic monophosphothioate, Sp-enantiomer (Sp-8Br-cAMPS), a cell membrane permeable activator of PKA, and three inhibitors of different serine/threonine protein phosphatases (okadaic acid, cantharidin, calyculin A), were tested. In RPMCs application of repetitive series of step hyper- and depolarizations (holding potential 0 mV, test potentials -80 to +80 mV, step size +20 mV) induced a slowly increasing, [half-maximal activation time ( t0.5) 11.0+/-1.1 min, Imax (at +80 mV) 18.7+/-3.1 pA pF-1], DIDS-sensitive, outwardly rectifying Cl- current I(Cl,OR). The activation of this current could be accelerated by Sp-8Br-cAMPS, okadaic acid or cantharidin in the extracellular solution. Co-application of Sp 8Br-cAMPS and okadaic acid increased Imax supra-additively. Calyculin A and higher concentrations of cantharidin inhibited the Cl- current via unknown mechanisms. Our findings suggest that I(Cl,OR) in RPMCs is activated by a PKA type protein kinase, a process which is antagonized functionally by okadaic acid- and cantharidin-sensitive protein phosphatases. PMID- 11907823 TI - Chronic hypoxia enhances endothelin-1-induced intracellular calcium elevation in rat carotid body chemoreceptors and up-regulates ETA receptor expression. AB - Endothelin-1 (ET-1) excites carotid body (CB) chemoreceptors and induces mitosis of the chemoreceptors in chronic hypoxia. The aim of the present study was to examine the hypothesis that up-regulation of both ETA receptor and endogenous ET 1 expression in CB chemoreceptors enhances the response of intracellular Ca2+ to ET-1 following adaptation to chronic hypoxia (10% inspired O2 for 3-4 weeks). Cytosolic free [Ca2+] ([Ca2+]i) in type-I (glomus) cells freshly dissociated from rat CBs was measured by spectrofluorometry. Application of exogenous ET-1 (1-100 nM) concentration-dependently elevated [Ca2+]i in the glomus cells. This response to ET-1 (100 nM) was 49% greater in the chronically hypoxic (CH) group. The ET-1 response was abolished completely by the ETA receptor antagonist BQ610 (1 microM), but not by the ETB antagonist BQ788 (1 microM). The transient [Ca2+]i elevation induced by caffeine (30 mM) in the normoxic group was similar to that in the CH group, suggesting no differences in the intracellular Ca2+ stores. In situ hybridization with a digoxigenin-labelled antisense ETA receptor mRNA oligonucleotide probe revealed very intense and ubiquitous specific expression of ETA receptors in the lobules of glomus cells in the CH group, whereas staining in normoxic controls was light. Immunohistochemical studies revealed intense cytoplasmic staining for ET-1-immunoreactivity in most of the cell clusters in glomera in the CBs of CH rats but was faint in normoxic CBs. These findings indicate increased expression of both the ETA receptor and ET-1 in CB chemoreceptors during chronic hypoxia. Taken together, our results suggest that the [Ca2+]i response to ET-1 in rat CB chemoreceptors is augmented by up regulation of ETA receptors and ET-1 expression. The enhancement of the paracrine/autocrine effect of ET-1 on the chemoreceptors is consistent with an excitatory and mitogenic role of the ET-1 and ETA receptor in the CB during chronic hypoxia. PMID- 11907824 TI - Effects of hydrogen peroxide and hydroxyl radicals on the cytosolic side of a non selective cation channel in the cultured human bronchial epithelial cell line 16HBE14o-. AB - Respiratory tissues can be damaged by the exposure of airway epithelial cells to reactive oxygen species (ROS) that generate an oxidative stress. We studied the effects of the hydroxyl radical *OH, for which there is no natural intra- or extracellular scavenger, on a Ca2+-activated, non-selective cation channel (NSC(Ca)) which might participate in the transepithelial Na+ fluxes involved in the maintenance of the cytosolic [Na+] ([Na+]i). We identified and characterized NSC(Ca) in inside-out excised membrane patches from cells of the human bronchial cell line 16HBE14o- and exposed the cytoplasmic side of NSC(Ca) to H2O2 or *OH created in front of the patch pipette. In these cells, the NSC channel was Ca2+ (above 0.1 microM) and voltage dependent. The channel's low open probability ( P(o)) at hyperpolarized voltages increased rapidly to a very high value upon short exposure (seconds) to *OH or to H2O2 in a bath solution containing millimolar [Ca2+]. This sensibility to ROS was reversible on wash-out of the oxidants. Long exposure (several minutes) to *OH in a bath solution containing 10(-7) M Ca2+ activated the channel irreversibly, a finding that has pathophysiological implications. The functioning of the NSC(Ca) protein channel may be modified by the intracellular redox status and the reversibility of the sensitivity to ROS depends on [Ca2+]i. The role of NSC(Ca) channels in airway epithelia is not yet understood, but might conform to what might be expected of an NSC channel: increase in [Na]i following intracellular exposure to *OH or H2O2. Reversible or irreversible excessive opening of cation channels may induce gradual cellular responses, including changes in Na+/K+-ATPase activity, cell volume regulation, membrane polarization, activation of transcription factors required for the expression of genes of cytokines or chemokines, apoptosis and necrosis, thus triggering inflammation. PMID- 11907825 TI - Change of quantal size and parameters of release with stimulation in bovine chromaffin cells. AB - Changes in quantal size and in the parameters of release were examined in chromaffin cells using amperometric recordings during and following various stimuli that induce secretion. As a general rule, a greater quantal content was associated with a greater quantal size. Following a short depolarizing pulse (0.5 2 s; 100 mV from a holding potential of -80 mV), the mean value of quantal size increased by 54% over several seconds before gradually (over tens of seconds) returning toward the control value, whilst its variability rose by 62%. The changes observed following 30-s applications of high extracellular K+ (50 mM) were more modest. A rapid application of short depolarizing pulses (2 s every 10 20 s; 100 mV from a holding potential of -80 mV) also led, at least initially, to greater quantal content and quantal size. Mean quantal size rose initially by 68%, but decreased subsequently to 31% below pre-stimulation levels. In whole clamped cells, the frequency of quantal release can rise abruptly, probably owing to a mechanical disturbance that makes the membrane leaky to Ca2+. In such cases, a marked rise in quantal content (>ten-fold) was paralleled by an almost as dramatic (up to ten-fold) rise in quantal size and an important, although less pronounced and slower, rise in its variability (up to four-fold). The return toward control values of mean quantal size occurred over several minutes, whilst its variability decayed more slowly. The release parameters were evaluated directly from the number of events to avoid a large and time-dependent contribution of the amplitude variability of spontaneous amperometric current spikes (minis). In general, the greater probability of release contributed more than the greater size of the immediately available store to the increase in quantal output. In conclusion, quantal size was found to be highly labile. Its change can alter strongly the facilitation and depression of evoked quantal output and probably occurs due to a preferential release of large vesicles that are more efficient barriers to Ca2+ diffusion when Ca2+ rises rapidly following a synchronous opening of several Ca2+ channels. When intracellular Ca2+ levels rise slowly to threshold levels for secretion, as during an asynchronous and generally spontaneous release, vesicles are less effective diffusion barriers and quantal size changes less. PMID- 11907826 TI - Nicotinic receptor/ionic channel complex (AChR) in androgen-dependent skeletal muscle cultures. AB - The kinetic properties of the nicotinic receptor/ionic channel complex (AChR) were compared in cell cultures obtained from androgen-dependent skeletal muscles of the perineal complex (P) and from muscles less dependent upon sex hormones (the thigh musculature, T). Because the development of P is delayed compared to other skeletal muscles in the rat, cultures were performed taking into account the age of the donor (4- or 6-day-old rats), and the time interval the cells remained in culture (7 days and 15 days). The ionic channel conductance (gamma) and the mean channel open time (tau) were determined with the patch-clamp technique in the cell-attached configuration at room temperature. Cultures from P and T muscles were morphologically identical in size and shape, independent of the animals' age at plating or on the plating time. In all of them, the AChR was spread over the cell membrane. More than one AChR ionic channel conductance was observed in P and T cultures, and the prevalent value of gamma in either culture ranged from 30 pS to 35 pS. In P fibers from 4-day-old rats cultured for 7 days (P 4/7), the distribution of channel open times fitted a double exponential, while in T 4/7 they were fitted with a single exponential. In cultures from P and T muscles obtained from older rats (6 days old) and in those cells remaining in culture for a prolonged time (15 days), the channel open times also fitted a double exponential. Because P and T cultures lack trophic neuronal influences, the difference observed between the tau of P 4/7 and T 4/7 was thought to be the hormone requirement of P muscles to grow and differentiate. Likewise, the difference observed between T 4/7 and T 4/15 may indicate the need for neurotrophic influences to maintain higher tau values in older cultures. Since this requirement is not found in cultured fibers, tau would tend to assume slower values approaching those of P without hormone activation. PMID- 11907827 TI - Divergent effects of NO synthase inhibition on systemic and myocardial O2 delivery and consumption during dobutamine infusion in sheep. AB - It is unknown if nitric oxide (NO) modulates the balance between systemic or myocardial O2 delivery ( DO2) and consumption VO2 during inotropic stimulation with the beta-adrenergic agonist dobutamine. Accordingly, we measured systemic and left ventricular (LV) DO2 and during dobutamine infusion to a peak of 10 microg .kg-1 .min-1, before and after inhibition of NO synthesis with i.v. N(omega)-nitro- L-arginine ( L-NNA, 25 mg.kg-1) in 11 anesthetized ewes instrumented with aortic, pulmonary arterial and coronary sinus catheters, an ultrasonic coronary artery flow probe and a pulmonary arterial thermistor to measure cardiac output. L-NNA reduced systemic DO2 by 19% (2.1+/-0.5 ml.min-1.kg 1, P<0.005) but increased systemic by 13% (0.4+/-0.1 ml.in-1.g-1, P<0.05), whereas LV DO2 and rose by 44% (4.3+/-1.3 ml.min-1).100 g-1, P<0.005) and 47% (3.1+/-0.9.min-1.100 g-1, P<0.025) respectively. Dobutamine increased systemic DO2 by 61% (5.3+/-0.9 ml.min-1.kg-1, P<0.001) and systemic by 10% (0.32+/-0.12 ml.min-1.kg-1, P<0.01), with no effect of L-NNA on either response ( P>0.9). As a result, while was 0.5 ml.min(-1).kg-1 higher at any given level of DO2 ( P<0.001), the slope of the systemic DO2- relation was unchanged after L-NNA. By contrast, LV DO2 increased by 152% (15.6+/-2.2 ml.min-1.100 g-1, P<0.001) and by 184% (12.7+/-1.9 ml.min-1.100 g-1, P<0.001), but because the slope of both dose response curves was reduced by 48% after L-NNA ( P<0.001), the LV DO2- relationship was unaffected by inhibition of NO synthesis. These results suggest that NO modulated the baseline balance between systemic, but not myocardial, DO2 and. However, NO did not additionally alter dobutamine-related changes in either systemic or myocardial DO2 and balance. PMID- 11907828 TI - Circadian variation in the effects of nitric oxide synthase inhibitors on body temperature, feeding and activity in rats. AB - We have investigated whether there is circadian variation in the effects of nitric oxide synthase inhibitors on body temperature, physical activity and feeding. We used nocturnally active Sprague-Dawley rats, housed at approximately 24 degrees C with a 12:12 h light:dark cycle (lights on 07:00 hours) and provided with food and water ad libitum. Nitric oxide synthesis was inhibited by intraperitoneal injection of the unspecific nitric oxide synthase inhibitor N nitro- L-arginine methyl ester ( L-NAME, 100, 50, 25, 10 mg/kg), or the relatively selective inducible nitric oxide synthase inhibitor aminoguanidine (100, 50 mg/kg), during the day ( approximately 09:00 hours) or night ( approximately 21:00 hours). Body temperature and physical activity were measured using radiotelemetry, while food intake was calculated by weighing each animal's food before as well as 12 and 24 h after each injection. We found that daytime injection of L-NAME and aminoguanidine had no effect on daytime body temperature. However, daytime injection of both drugs did decrease nocturnal food intake ( P<0.05) and activity ( P<0.05). When injected at night, L-NAME reduced night-time body temperature ( P<0.01), activity ( P<0.05) and food intake ( P<0.05) in a dose-dependent manner, but night-time injection of aminoguanidine inhibited only night-time activity ( P<0.05). The effects of nitric oxide synthase inhibition on body temperature, feeding and activity therefore are primarily a consequence of inhibiting constitutively expressed nitric oxide synthase, and are subject to circadian variation. PMID- 11907829 TI - Cerebral localization and regulation of the cell volume-sensitive serum- and glucocorticoid-dependent kinase SGK1. AB - The serum- and glucocorticoid-dependent kinase SGK1 is regulated by alterations of cell volume, whereby cell shrinkage increases and cell swelling decreases the transcription, expression and activity of SGK1. The kinase is expressed in all human tissues studied including the brain. The present study was performed to localize the sites of SGK1 transcription in the brain, to elucidate the influence of the hydration status on SGK1 transcription and to explore the functional significance of altered SGK1 expression. Northern blot analysis of human brain showed SGK1 to be expressed in all cerebral structures examined: amygdala, caudate nucleus, corpus callosum, hippocampus, substantia nigra, subthalamic nucleus and thalamus. In situ hybridization and immunohistochemistry in the rat revealed increased expression of SGK1 in neurons of the hippocampal area CA3 after dehydration, compared with similar slices from brains of euvolaemic rats. Additionally, several oligodendrocytes, a few microglial cells, but no astrocytes, were positive for SGK1. The abundance of SGK1 mRNA in the temporal lobe, including hippocampus, was increased by dehydration and SGK1 transcription in neuroblastoma cells was stimulated by an increase of extracellular osmolarity. Co-expression studies in Xenopus laevis oocytes revealed that SGK1 markedly increased the activity of the neuronal K+ channel Kv1.3. As activation of K+ channels modifies excitation of neuronal cells, SGK1 may participate in the regulation of neuronal excitability. PMID- 11907830 TI - IGF-1 up-regulates K+ channels via PI3-kinase, PDK1 and SGK1. AB - Involvement of voltage-gated (Kv) potassium channels in IGF-1-induced proliferation of HEK293 cells was studied by patch-clamp, RT-PCR and FACS analysis. IGF-1 up-regulated outwardly rectifying whole-cell K+ current starting after 1 h of incubation and reaching a maximum after 4-6 h. The IGF-1-stimulated current was voltage-gated with an activation threshold of -30 mV to -40 mV, a half-maximal activation at +5.3+/-1.8 mV, and time constants for activation and inactivation of 4.5+/-0.4 ms and 43.5+/-5.6 ms ( n=10), respectively. The current was inhibited by TEA, margatoxin, agitoxin-2 and stichodactyla toxin. PCR amplification of different Kv subunits from HEK293 cDNA demonstrated the expression of Kv1.1, Kv1.2, Kv1.3, Kv1.4, Kv3.1 and Kv3.4 mRNA. Quantitative RT PCR showed up-regulation of Kv1.1, 1.2 and 1.3 mRNA by IGF-1. The effect of IGF-1 on K+ current was blocked by inhibitors of phosphatidylinositol 3-kinase (PI3 kinase), wortmannin and LY294002, and mimicked by overexpression of human 3 phosphoinositide-dependent protein kinase-1 (hPDK1) or serum- and glucocorticoid dependent kinase-1 (hSGK1), both sequential downstream targets of PI3-kinase. IGF 1-induced proliferation of HEK293 cells was inhibited by both K+ channel blockers and inhibitors of PI3-kinase. In conclusion, IGF-1 through PI3-kinase, PDK1 and SGK1 up-regulates Kv channels, an effect required for the proliferative action of the growth factor. PMID- 11907831 TI - Opioid receptor-mediated effects of interleukin-2 on the [Ca2+]i transient and contraction in isolated ventricular myocytes of the rat. AB - Cytokines play significant roles in some cardiovascular disorders, but direct myocardial effects of cytokines remain to be elucidated. In this study, we examined the effects and possible mechanisms of interleukin-2 (IL-2) on contraction and the [Ca2+]i transient of enzymatically isolated ventricular myocytes with spectrofluorometry and video tracking. IL-2 (2.5-200 U/ml) depressed both the contraction and the [Ca2+]i transient in a dose-dependent manner. Pretreatment with the universal opioid receptor antagonist naloxone (10 nM), or a specific kappa opioid receptor antagonist, nor-binaltorphimine (nor BNI, 10 nM), abolished the inhibitory effect of IL-2 on contraction and the [Ca2+]i transient; the specific delta-opioid receptor antagonist naltrindole (1 microM) had no effect. The effect of IL-2 was also abolished after pretreatment with pertussis toxin (PTX, 5 mg/l), but not by genistein (100 microM). Pretreatment with the phospholipase C inhibitor U73122 (5 microM) significantly inhibited the IL-2-induced depression of contraction and the [Ca2+]i transient. It is concluded that the effects of IL-2 on contraction and the [Ca2+]i transient of ventricular myocytes are mediated via the cardiac kappa opioid receptor, and the postreceptor signal transduction pathway includes a PTX-sensitive G protein and phospholipase C. PMID- 11907832 TI - Theoretical estimation of the capacity of intracellular calcium stores in the Bergmann glial cell. AB - We propose a mathematical model for calcium dynamics ([Ca2+]i) during metabotropic activation of a specialized astrocyte, the cerebellar Bergmann glial cell. The model adequately describes the experimentally observed behaviour of the prototype in response to single and repetitive metabotropic stimuli and to the inhibition of Ca2+ uptake into the store. By means of the model, the capacity of the intracellular calcium store for two types of calcium buffer was estimated. The estimated buffer capacity of the store lies within the following intervals: (0.8-15.5).10-19 mol calmodulin, and (0.6-12.3).10-19 mol calbindin. This result reveals, that, in the store of a small Ca2+-containing compartment optically detected in the Bergmann glial cell process after electrical stimulation of parallel fibres, the amount of releasable Ca2+ does not exceed 25,000 ions. The quantitative estimates were obtained from experimentally based theoretical relationships between the capacity and volume of the store and parameters of the cytoplasmic calcium buffer. In these relationships, the estimated store capacity was proportional to the total buffer concentration, inversely proportional to the constant of buffer affinity for calcium and was smaller for a greater relative store volume. PMID- 11907834 TI - Membrane capacitance measurement using patch clamp with integrated self-balancing lock-in amplifier. AB - A lock-in amplifier was incorporated directly into the resistance and capacitance compensation circuitry of a patch clamp set-up, to allow exocytosis to be monitored in the whole-cell mode by measuring changes in cell membrane capacitance. The integration of these two systems enabled us to provide a novel operating mode, which we term the "track-in" mode, where the output signals from the lock-in amplifier are used to make compensating electronic adjustments of the resistance and capacitance control settings. The lock-in amplifier outputs remain near zero, and the control voltages generated by the feedback circuits provide linear and calibrated resistance and capacitance measurements. Results obtained from model cells and mouse inner hair cells show that this is achieved without loss of sensitivity or of significant time resolution compared with the conventional lock-in amplifier technique, whereas the sensitivity to switching phase errors is effectively eliminated. An automatic phase tracking system using a low-frequency dithering of the resistance control setting can nevertheless be used to preserve the correct switching phase if required. The track-in approach has considerable advantages over software implementations in terms of economy and convenience, since the resistance and capacitance signals can be recorded directly on any general-purpose data-acquisition system. PMID- 11907835 TI - Single-cell electroporation. AB - Using modified patch-clamp methodology, we demonstrated that it is possible to insert genes or other compounds routinely into single cells by electroporation. When the cell is indented by a small-tipped microelectrode, a voltage of 10 V or less in the pipette is divided by the pipette resistance and the series resistance of the cleft between the pipette tip and the cell surface. The voltage at the cell membrane can be high enough to cause localized dielectric breakdown of the membrane and create pores that allow compounds in the pipette to enter the cell. Rectangular pulses from 20 micros to more than 300 ms are effective, as are frequencies from DC to 5 kHz. The most significant parameter was the total time for which the voltage was applied. Pipette voltages of 2-10 V were required, with larger genes requiring larger voltages. With optimal parameters, transfection rates in excess of 80% were also possible routinely. This approach offers an effective alternative to intracellular pressure injection and iontophoresis for placing genes, drugs, and other compounds in cells. Because of the small size of the electrode tips, substances can be inserted in cells from almost any location on their surfaces. In addition, the small tips electroporated only a limited area and so did little cell damage. PMID- 11907837 TI - Dermatological manifestations of Down's syndrome. AB - BACKGROUND: Down's syndrome (DS) is an extensively researched congenital condition characterized by mental retardation and distinct physical features. The cutaneous manifestations of DS are numerous, yet they seldom receive appropriate attention. OBJECTIVE: To review the dermatological conditions associated with DS. METHODS: A review of the medical literature. RESULTS: DS is associated with an increased incidence of numerous dermatological conditions, some of which may be related to an immunological deficiency. CONCLUSION: Along with the importance of understanding the physical and psychosocial aspects of DS, an appreciation of the dermatological manifestations of this condition is integral to comprehensive medical care. PMID- 11907838 TI - Ultraviolet light (UV) regulation of the TNF family decoy receptors DcR2 and DcR3 in human keratinocytes. AB - BACKGROUND: Several additional members of the tumor necrosis factor (TNF) receptor family were recently identified. The existence of such receptors, which may play distinct and unique regulatory roles, suggests that complex regulatory mechanisms are involved in apoptosis. OBJECTIVE: This study examines the expression of several members of the TNF receptor family in human keratinocytes exposed to ultraviolet B (UVB) irradiation. METHODS: Human keratinocytes were exposed to increasing doses of UVB, total RNA was harvested, and a quantitative RNase protection assay was performed. RESULTS: Decoy receptor-3 (DcR3), a nonfunctional receptor that binds to Fas ligand (FasL), was constitutively expressed at high level in keratinocytes but decreased rapidly in cells exposed to UVB. Decoy receptor-2 (DcR2), a nonfunctional receptor that binds to TNF related apoptosis-inducing ligand (TRAIL)/APO-2L, showed the opposite expression pattern. DcR2 was undetectable in unirradiated keratinocytes and was markedly up regulated after exposure to UVB. Although the response showed significant delays at higher UVB doses, the patterns observed for DcR3 and DcR2 were consistent in this set of experiments. CONCLUSION: We conclude that UVB regulates expression of these two TNF decoy receptors in keratinocytes. This pathway may represent a novel mechanism for regulation of apoptosis in the skin. PMID- 11907839 TI - Management of psoriasis vulgaris with methotrexate 0.25% in a hydrophilic gel: a placebo-controlled, double-blind study. AB - BACKGROUND: Methotrexate has been used as one of the first and systemic therapies for psoriasis. In general, 70% of patients with psoriasis prefer topical therapy as the treatment of choice. OBJECTIVE: The purpose of this placebo-controlled double-blind study was to evaluate the clinical efficacy and tolerability of methotrexate 0.25% incorporated in a hydrophilic gel (hydroxyethylcellulose 1%) to treat patients afflicted with psoriasis vulgaris. METHODS: Sixty patients (37M/23F) ranging between 18 and 70 years of age, with slight to moderate chronic plaque-type psoriasis and PASI (Psoriasis Area and Severity Index) scores between 5.3 and 17.5 joined the study. The mean duration of the disease at entry was 9.6 years (range 1-24 years). The diagnosis of psoriasis was established by clinical and histopathologic methods. Patients were sequentially randomized into two parallel groups. Each patient was allocated a precoded 100-g tube (active or placebo) with instructions on how to self-administer the trial medication topically (without occlusion) to their lesions two times daily for 5 consecutive days per week. The study lasted for 12 weeks with 4 weeks of active treatment. Patients were examined on a weekly basis and those showing total clearing or remission of lesions were considered effectively treated. RESULTS: By the end of the treatment, breaking the code disclosed that methotrexate 0.25% gel had significantly treated more patients than placebo (83.3% vs. 6.7%; p < 0.0001), reduced the PASI score to a mean of 2.2, and cleared more plaques (82.2% vs. 4.3%; p < 0.0001). Laboratory evaluations, including CBC with differential and platelet count, renal function, liver chemistry [SGOT (aspartate transaminase) and SGPT (alanine transaminase)], and serum creatinine, were within the normal limits. The treatment was well-tolerated by all the patients, with no adverse drug-related symptoms and no dropouts. The study was followed up for 12 months from the first day of the treatment; two cured patients had relapsed after 8 months. CONCLUSION: The findings of this study demonstrate that methotrexate 0.25% in a hydrophilic gel is well tolerated and significantly more effective than placebo as a patient-applied topical medication to treat psoriasis vulgaris. PMID- 11907840 TI - Betamethasone valerate foam for treatment of nonscalp psoriasis. AB - BACKGROUND: Although betamethasone valerate (BMV) foam, 0.12% (Luxiq, Connectics Corporation, Palo Alto, CA) is approved by the Food and Drug Administration for the treatment of corticosteroid-responsive scalp dermatoses, no data are available for its use on nonscalp psoriasis. OBJECTIVE: We evaluated the safety and efficacy of BMV foam in treating psoriatic lesions at nonscalp sites. METHODS: We conducted a randomized, double-blind, placebo-controlled, paired comparison, split-body study of 40 patients with mild to moderate plaque-type psoriasis. Patients applied BMV foam and placebo foam twice daily for 12 weeks. RESULTS: At the end of the treatment period, 70% of patients had greater than 50% improvement of lesions on their active foam-treated side compared with 24% of patients with similar improvement on their placebo foam-treated side. Adverse effects were limited to temporary stinging, burning, or itching in several patients. Three patients (7.5%) withdrew because of stinging or itching. CONCLUSIONS: The results indicate that BMV foam is effective against nonscalp psoriasis. Twice-daily applications are well tolerated, compliance exceeds 90%, cosmetic characteristics are acceptable, and the medication may reduce the need for multiple prescriptions. PMID- 11907841 TI - Bone lesions and facial papules in a 41-year-old female. PMID- 11907842 TI - Arthropod bites manifesting as recurrent bullae in a patient with chronic lymphocytic leukemia. AB - BACKGROUND: We report a patient with chronic lymphocytic leukemia (CLL) that developed recurrent vesicobullous lesions that histologically demonstrated features of an exaggerated response to an arthropod bite. OBJECTIVE: Patients with CLL can present with many cutaneous manifestations, including specific and nonspecific lesions. Although rare, patients with CLL can develop an exaggerated response to an arthropod bite. CONCLUSION: Emphasis needs to be placed on the clinical recognition of arthropod bites as an unusual cutaneous manifestation of CLL, as they provide the physician with both a diagnostic and a therapeutic challenge. Patients often deny being bitten and, thus, the biopsy results conflict with the patient's history. Additionally, as there is no specific treatment, both the patient and physician are faced with a similar dilemma. Although our patient initially responded well to corticosteroids, his lesions significantly improved while being treated with dapsone. PMID- 11907843 TI - Peroxisomes in dermatology. Part II. AB - BACKGROUND: Peroxisomes are small cellular organelles that were almost ignored for years because they were believed to play only a minor role in cellular functions. However, it is now known that peroxisomes play an important role in regulating cellular proliferation and differentiation as well as in the modulation of inflammatory mediators. In addition, peroxisomes have broad effects on the metabolism of lipids, hormones, and xenobiotics. Through their effects on lipid metabolism, peroxisomes also affect cellular membranes and adipocyte formation, as well as insulin sensitivity, and peroxisomes play a role in aging and tumorigenesis through their effects on oxidative stress. OBJECTIVE: To review genetically determined peroxisomal disorders, especially those that particularly affect the skin, and some recent information on the specific genetic defects that lead to some of these disorders. In addition, we present some of the emerging knowledge of peroxisomal proliferator activator receptors (PPARs) and how ligands for these receptors modulate different peroxisomal functions. We also present information on how the discovery of PPARs, and the broad and diverse group of ligands that activate these members of the superfamily of nuclear binding transcription factors, has led to development of new drugs that modulate the function of peroxisomes. CONCLUSION: PPAR expression and ligand modulation within the skin have shown potential uses for these ligands in a number of inflammatory cutaneous disorders, including acne vulgaris, cutaneous disorders with barrier dysfunction, cutaneous effects of aging, and poor wound healing associated with altered signal transduction, as well as for side effects induced by the metabolic dysregulation of other drugs. PMID- 11907844 TI - Human papillomavirus, smoking, and cancer. AB - BACKGROUND: The effect of smoking on human papillomavirus (HPV) activity and subsequent dysplasia and neoplasia remains controversial. OBJECTIVE: To determine any reported effects of smoking on either HPV activity or HPV-related dysplasia/cancer using retrospective analysis of the literature from 1966 through 1998 via Toxline and PubMed to search for "smoking," "papillomavirus," and "cancer." CONCLUSION: Several recent large studies demonstrated that smoking was associated with a greater incidence of cervical, vulvar, penile, anal, oral, and head and neck cancer in a dose-dependent fashion, while other studies did not show any correlation between smoking and cervical dysplasia after multivariate adjustment. Recent studies have also indicated that smoking may be more closely related to high-grade lesions of the cervix and vulva. These data provide evidence of an association between HPV, smoking, and cancer. Progression of dysplasia likewise seems to be associated with smoking. Several groups have attempted to discern whether the connection between smoking and cervical cancer is from local immunosuppression and/or from direct carcinogenic effects. PMID- 11907845 TI - Tacrolimus in dermatology. AB - BACKGROUND: Tacrolimus (FK 506), a metabolite of the fungus Streptomyces tsukubaensis, is an anti-T-cell drug. It acts by inhibiting the production of IL 2, IL-3, IL-4, TNFa, and GM-CSF. More potent and with slightly less secondary effects than cyclosporine, it has been the object of considerable interest, especially in conditions that could benefit from the latter. OBJECTIVE: In psoriasis, a placebo-controlled double-blind study has shown oral tacrolimus at 0.1 mg/kg/day to be effective in controlling recalcitrant lesions. In human, small studies have reported tacrolimus ointment to be effective in controlling acute contact dermatitis. Short-term trials of topical tacrolimus in the treatment of atopic dermatitis have recently shown excellent results in both adults and children. In animal studies of hair growth disorders, topical tacrolimus induces anagen and protects from chemotherapy-induced alopecia. Animal studies with the ointment for the prevention of skin graft rejection, lupus dermatoses, and skin papilloma formation have also shown to be promising. CONCLUSIONS: There are case reports of pyoderma gangrenosum, Sezary's syndrome, and Behcet's disease successfully treated with oral tacrolimus but, because of their small number, they remain anecdotal at this point. PMID- 11907846 TI - Dural melanoma associated with ocular melanosis and multiple blue nevi. AB - BACKGROUND: Primary meningeal melanomas of the central nervous system (CNS) are a rare malignant process with the majority originating from the leptomeninges. Primary dural melanomas have been reported to occur in isolation or in conjunction with Nevus of Ota. The association of primary dural melanoma with multiple cutaneous blue nevi has not been reported previously. OBJECTIVE: To describe a case of a 41-year-old Asian woman patient with a primary dural melanoma that arose in association with ocular melanosis and multiple cutaneous blue nevi. The patient is alive almost more than 8 years after subtotal and subsequent total resection of her primary tumor. Primary dural melanomas, Nevus of Ota, and blue nevi are discussed in relation to their coexistence and potential for intracranial melanoma. CONCLUSION: CNS melanoma is regarded as an extremely aggressive disease with poor prognosis. This case and previous reports of dural melanomas occurring in isolation or with Nevus of Ota have demonstrated relatively prolonged survival after surgical intervention. We conclude that dural melanomas are less aggressive tumors requiring surgical extirpation only. PMID- 11907848 TI - Microscopic acanthosis nigricans in type 2 diabetes. AB - BACKGROUND: Acanthosis nigricans (AN) has been associated with insulin resistance. Individuals with type 2 diabetes are insulin-resistant and, therefore, could be expected to manifest AN. However, the prevalence and predictors of AN are unknown in this population. OBJECTIVE: An outpatient population with Type 2 diabetes (DM) was compared with matched controls (C) for microscopic and clinical AN along with measurement of body habitus, insulin, glucose, and androgen levels. METHODS: Twenty-four individuals with DM (12M, 12F) from a tertiary care center were compared with 24 C (12M, 12F). Fasting glucose, insulin, sex hormone binding globulin, androstenedione, dihydroepiandrosterone sulfate, and testosterone were measured. Height, weight, waist/hip measures, and a clinical survey for acanthosis were recorded. A 2-mm skin biopsy from midaxilla of the nondominant arm was taken for pathological review. RESULTS: C and DM were matched for age and body mass index (BMI). Prevalence of microscopic AN in C was 12% (3/24) and in DM was 21% (5/24; NS). In C, AN was predicted by waist, waist/hip ratio, and fasting insulin measures, while none of the variables examined was predicative of AN in DM. CONCLUSIONS: Microscopic acanthosis nigricans was found in similar numbers of people with DM when compared with C. Fasting insulin levels most strongly predicted the presence of AN in C, while no significant predictors of AN were found in the population with DM. PMID- 11907847 TI - Betamethasone valerate in foam vehicle is effective with both daily and twice a day dosing: a single-blind, open-label study in the treatment of scalp psoriasis. AB - BACKGROUND: Psoriasis is a chronic relapsing skin disorder that affects about 2% of the U.S. population and involves the scalp approximately 50% of the time. Topical corticosteroids, including betamethasone valerate, have been used effectively in the treatment of corticosteroid-responsive dermatoses of the skin and scalp. Betamethasone valerate (BMV) in foam vehicle (Luxiq) is designed to improve patient compliance with topical therapy. Superior efficacy over a BMV lotion preparation has been demonstrated with twice-daily use. Even greater compliance would be expected if the drug is effective with once-daily application. PURPOSE: To compare the efficacy of the betamethasone valerate foam (Luxiq) in the treatment of scalp psoriasis following once-daily versus twice daily dosing. METHODS: Seventy-nine patients with moderate to severe scalp psoriasis from seven centers were enrolled and treated with BMV foam either once a day or twice a day for four weeks. The physician-grader was blinded to the treatment regimen, and the subjects were randomly assigned to either once-daily or twice-daily dosing in a 1:1 ratio. RESULTS: The signs of psoriasis (plaque thickness, scaling, and erythema) were assessed before and after treatment. The investigator's and the patients' global assessments were also evaluated. The composite score improved from 7.7 +/- 2.1 to 3.0 +/- 2.2 with twice-a-day use and from 8.1 +/- 2.2 to 3.9 +/- 2.8 with once-daily use (p > 0.05 for the difference between groups). DISCUSSION: BMV foam is effective for scalp psoriasis with both once-a-day and twice-a-day use. This feature of the BMV foam is encouraging for expected improvement in clinical use. PMID- 11907849 TI - Bullous pilomatricoma: a report of clinical and pathologic findings and review of dermal bullous disorders. AB - BACKGROUND: Pilomatricoma is a common benign adnexal tumor differentiating toward elements of the hair matrix and shaft. It typically presents as a solitary, deep, dermal nodule. We describe a case of a pilomatricoma with the unusual feature of a thick-walled dermal bulla overlying the tumor. OBJECTIVE: We describe a case of bullous pilomatricoma and discuss the potential etiology of the bullous feature of the lesion. METHODS: This article includes a case report and a literature review. CONCLUSIONS: Bullous pilomatricoma has rarely been described. A common pathological feature in this type of pilomatricoma is the presence of dilated lymphatics. Bullous morphea associated with dermal lymphatic dilation has also been described. In both bullous pilomatricoma and morphea, it is possible that individual pathological features of the lesion lead to obstruction and congestion of the dermal lymphatics thereby inducing enough dilation and edema to form a dermal bulla. PMID- 11907850 TI - Epidermolysis bullosa acquisita responsive to dapsone therapy. AB - BACKGROUND: Epidermolysis bullosa acquisita (EBA) is a chronic subepidermal blistering disease that is frequently resistant to therapy. OBJECTIVE: A 58-year old man who had a one-year history of a bullous eruption involving the hands, forearms, trunk, scalp, and oral mucosa. Histopathology revealed a subepidermal bulla, and direct and indirect immunofluorescence studies were consistent with EBA. The patient failed respond to niacinamide and tetracycline and oral prednisone 40 mg per day. METHODS: Complete control of his blistering was achieved within two months of initiating oral dapsone, 150 mg per day. CONCLUSION: Dapsone may be an effective agent for some patients with EBA. PMID- 11907851 TI - Are cutaneous reactions to fly larvae mediated by CD4+, TIA+ NK1.1 T Cells? AB - BACKGROUND: Although there have been reports of fly larvae in wounds and as isolated primary infestations, there have been only rare reports documenting reactions to the larvae within the skin in humans and animals. There have been no reports documenting the histopathologic and immunohistochemical characteristics of the inflammatory infiltrate. OBJECTIVE: We present a patient who developed local pruritus, erythema, and swelling approximately three weeks after infestation by a fly larva within the scalp. Histopathologically the biopsy site showed a mixed infiltrate containing lymphoid cells and numerous eosinophils. Immunohistochemical stains showed predominantly CD4+ T cells expressing an ab T cell receptor (TCR) of which approximately 30% coexpressed T-cell intracellular antigen (TIA) and CD56. In addition, there were approximately 5% of these CD4+ T cells which coexpressed CD30. CONCLUSIONS: Histopathologic and immunohistochemical findings are consistent with an effector cell population of cytotoxic CD4+ T cells that produce a T-helper 2 cytokine pattern. The phenotype of this subset of T cells is unique and among its characteristics is that antigens--usually nonprotein antigens--are presented to these CD4+,TIA+ natural killer (NK)1.1T cells by CD1d molecules. PMID- 11907852 TI - Reduction of postherpetic neuralgia in herpes zoster. AB - BACKGROUND: Persons 50 years of age and older are not only at increased risk of developing herpes zoster, they are also more likely to suffer the long-term morbidity of postherpetic neuralgia (PHN). PHN is pain persisting after the rash of herpes zoster has healed. PHN affects at least 40% of all herpes zoster patients over age 50 and over 75% of herpes zoster patients over age 75; PHN is the single most common neurologic condition in elderly patients. OBJECTIVE: The objective of this review is to evaluate interventions that may reduce or even eliminate PHN. No single therapy has been consistently effective for PHN. The most effective approach appears to be with the use of antiviral therapy early in the course of herpes zoster. The goals of ongoing studies in herpes zoster are to develop interventions that will further reduce the symptoms of PHN and/or to eliminate PHN by prophylaxis using the varicella vaccine. CONCLUSIONS: Reduction of PHN can best be achieved with the use of antiviral medication early in the course of herpes zoster; other classes of drugs are minimally effective in treating established PHN. Widespread use of the varicella vaccine may lead to secondary reductions in PHN in the distant future. PMID- 11907853 TI - Pyogenic granuloma: pyogenic again? Association between pyogenic granuloma and Bartonella. AB - BACKGROUND: Pyogenic granulomas (PG) are benign vascular lesions which were thought to have an infectious etiology, yet none has been found. Bacillary angiomatosis (BA), which presents as disseminated vascular lesions in immunosuppressed patients, and verruga peruana (VP), which presents as crops of vascular nodules in immunocompetent persons, are caused by infection with Bartonella. Thus, the question was raised whether Bartonella could be associated with the development of PG, also a vasoproliferative lesion like BA and VP. The objective of this study was to determine through a case-control study whether such an association exists. METHODS: Patients who presented with PG and age and sex-matched controls with capillary hemangiomas and senile (cherry) angiomas were tested for serum IgG antibodies against Bartonella using an immunofluorescence antibody method. The prevalence of positive serology was compared between the groups. RESULTS: Twenty PG patients and 20 control patients with hemangiomas or angiomas were studied. Six out of 20 PG patients tested positive (30%), while none of the 20 control patients tested positive (0%). The difference between the proportions of seropositivity in the two groups reached statistical significance (p = 0.02, df = 1). CONCLUSIONS: Pyogenic granuloma patients were determined to have a statistically higher prevalence of Bartonella seropositivity compared with control patients. Further studies are needed to confirm the association and establish a possible etiological link. Such an association could have potential therapeutic importance. A nonsurgical approach with antibiotics may be possible and may decrease the recurrence rate and occurrence of satellite lesions. PMID- 11907854 TI - Attitudes of female patients regarding oral contraceptives for treatment of acne. AB - BACKGROUND: Two hormonal agents with contraceptive properties are currently indicated in Canada for treatment of acne. OBJECTIVE: The purpose of this study was to determine the attitudes and concerns of female patients regarding the use of hormonal agents and oral contraceptives in treatment of acne. METHODS: A self completed questionnaire was administered to all female acne patients of a community-based dermatologist over a 10-week period between June and August 1999. Statistical analysis was performed by chi-square testing at the 5% significance level. RESULTS: Eighty-six percent recognized that oral contraceptives ("birth control pills") were also used as a treatment for acne and 60% were willing to use this form of treatment. The most common concern regarding the use of oral contraceptives were side effects (80%). Younger patients were seven times more likely to be concerned about potential disapproval or misunderstanding on the part of others regarding the use of these medications (p approximately 0.01). CONCLUSION: Female acne patients recognize that oral contraceptives are useful in treatment of acne. Although side effects of these medications are of general concern, social disapproval is of particular concern to younger patients. PMID- 11907855 TI - Toxic epidermal necrolysis of the scalp following anticonvulsant use and cranial irradiation. AB - BACKGROUND: Toxic epidermal necrolysis (TEN) of the scalp is rare but it has been shown to occur in patients who had been given a combination of cranial radiation and anticonvulsant therapies. OBJECTIVE: We present a 62-year-old man who received cranial irradiation following craniotomy for glioblastoma multiforme. After he was prescribed the anticonvulsant phenytoin for postsurgical seizure prophylaxis, the patient developed TEN which began on the scalp before spreading to involve other parts of his body. Our second case was a 55-year-old woman who had been diagnosed with lung carcinoma with metastasis to the brain. She was treated with cranial irradiation and the anticonvulsant carbamazepine. TEN developed first on the scalp and then became generalized. CONCLUSIONS: While the combination of radiation and anticonvulsants leads to an increased risk of developing TEN, cranial irradiation appears to be the localizing factor in the development of TEN of the scalp. PMID- 11907856 TI - Pseudoporphyria: an atypical variant resembling toxic epidermal necrolysis. AB - BACKGROUND: Pseudoporphyria has been attributed to both medication usage and chronic hemodialysis. Histologically, it is identical to porphyria cutanea tarda. It is most commonly seen as localized bullae on sun-exposed skin, often on the dorsum of the hands and fingers. OBJECTIVES: We describe a 31-year-old man with rapidly evolving bullae which became denuded, clinically suggestive of toxic epidermal necrolysis. Pseudoporphyria was proven histologically. However, our patient's eruption was not localized as small bullae but was widespread, with large bullae evolving into large, cutaneous, denuded erosions. CONCLUSIONS: We describe a previously unreported, generalized variant of pseudoporphyria that resembles toxic epidermal necrolysis. We provide a review of pseudoporphyria and compare our variant to toxic epidermal necrolysis and mimicking disorders. PMID- 11907857 TI - Self-healing congenital Langerhans cell histiocytosis presenting as neonatal papulovesicular eruption. AB - BACKGROUND: Congenital self-healing Langerhans cell histiocytosis (CSHLCH) is a rare condition which may present at birth or during the neonatal period. It is usually characterized by the eruption of multiple, disseminated, red-brown papules and nodules which may increase in size and number during the first few weeks of life. Systemic signs are usually absent except for occasional mild hepatomegaly. OBJECTIVE: We present a 3.5-kg male infant who presented at birth with numerous diffuse, erythematous, crusted erosions. He was presumed to have congenital herpes simplex virus (HSV) and was started on IV acyclovir. Histopathologic examination revealed a mixed inflammatory infiltrate with numerous histiocytes which were S-100 and peanut agglutin positive consistent with CSHLCH. Further workup did not reveal any signs of systemic involvement. CONCLUSION: CSHLCH has rarely been reported to present as a papulovesicular eruption at birth. In these cases, a viral etiology is commonly entertained in the differential diagnosis. Despite the spontaneous regression of skin lesions in CSHLCH, close followup is required to evaluate for systemic signs and symptoms associated with latent Letterer-Siwe disease. PMID- 11907858 TI - Multiple sebaceous adenomas and extraocular sebaceous carcinoma in a patient with multiple sclerosis: case report and review of literature. AB - BACKGROUND: Sebaceous carcinomas are relatively rare cutaneous tumors; there are fewer than 600 cases reported. They most commonly arise within the meibomian gland of the eyelid; fewer than 150 of the reported cases were extraocular. These tumors have a high incidence of local recurrence and regional metastasis. The relationship of sebaceous tumors and visceral malignancy is well established in the literature. OBJECTIVE: We describe a 44-year-old white female with chronic progressive multiple sclerosis who developed multiple scalp sebaceous adenomas and a solitary sebaceous carcinoma. CONCLUSION: Extraocular sebaceous carcinomas are rare tumors with high incidence of local recurrence and regional metastasis. Surgery with wide surgical margins is the standard treatment. We report the first case of multiple sebaceous tumors in a patient with multiple sclerosis. The presence of sebaceous tumors warrants a search for an internal malignancy. Literature on sebaceous tumors and their relationship with visceral malignancies and immunologic disorders is discussed. Literature on sebaceous carcinoma with special attention to extraocular sebaceous carcinoma is also discussed. PMID- 11907859 TI - Review of ivermectin in scabies. AB - BACKGROUND: Currently available topical medications for scabies are messy and need prolonged application. This leads to poor patient adherence. Emerging drug resistance to topical scabicides has made eradication of scabies difficult. OBJECTIVE: Availability of an effective oral scabicidal agent, ivermectin (Ivm), opens a new era in the management of scabies. This review summarizes the published literature on the use of ivermectin in the treatment of scabies. CONCLUSION: A single oral dose of ivermectin of 200 mg/kg body weight is very effective in the treatment of human scabies. A second dose 7-10 days later substantially improves the cure rate. This suggests that ivermectin may not be effective against all the stages in the life cycle of the parasite. Endemic and epidemic scabies in institutions are better treated with Ivm. Crusted scabies ideally should be treated with a combination of Ivm and topical scabicides. Other standard measures and precautions should be taken. Additional controlled studies using a higher single oral dose or using parenteral or topical forms of Ivm are needed. The safety of Ivm in children less than 5 years old and in pregnant women has to be established. The U.S. FDA has not yet approved the drug for the treatment of human scabies. PMID- 11907860 TI - What's in a name? Comments on the dermatological dictionary by Ledier, Rosenblum, and Carter. AB - BACKGROUND: Any scientific discipline needs a sharply defined set of words for exact and reproducible communication. Surprisingly this has never been a strong point in dermatological science, especially as regards the living gross pathology of skin disease. OBJECTIVE: This article briefly reviews the four editions of a dermatological dictionary of words and phrases and gives some thoughts on their usefulness. CONCLUSION: My conclusions are twofold: we need a new dictionary; at the very least, we need a reprinting of the fourth edition of "A Dictionary of Dermatologic Terms" by Carter. PMID- 11907861 TI - [Treatment of obesity in practice - more necessary than ever]. PMID- 11907862 TI - [Sibutramine in clinical practice - a PMS-study with positive effects on blood pressure and metabolic parameters]. AB - BACKGROUND AND AIM OF THE STUDY: According to the latest health-survey data of the Robert Koch Institute, more than a fifth of all Germans are obese and about two thirds are overweight. As non-pharmacological therapies have failed to a large extent in the past, this post-marketing surveillance study was conducted to examine the feasibility, safety and efficacy of a medical weight reduction. PATIENTS AND METHODS: 6360 patients with a BMI of at least 30 kg/m(2) or, in the case of further risk factors, of 27 kg/m(2) or more, were included in the trial; the inclusion criteria and exclusion criteria chosen adhered strictly to the directions of use given for sibutramine. Two thirds of the patients had concomitant diseases and/or concomitant risk factors, and 62.4 % used concomitant medications. After being informed about non-medical therapies, the patients received 10 mg sibutramine once daily. After four weeks this treatment could be continued, terminated or increased to a dosage of 15 mg sibutramine per day. RESULTS: The body weight was reduced from 98.4 kg to 88.4 kg within the 12 weeks of treatment, which corresponds to a mean reduction of the BMI of 3.7 kg/m (2) and which was accompanied by clinically relevant reductions of waist and hip measurements. The following metabolic parameters improved significantly: cholesterol, LDL, HDL, triglycerides, blood glucose, HbA1c and uric acid. Whereas the blood pressure of normotensive patients was unchanged during the course of the study, or increased very slightly, for patients presenting with hypertension at baseline a significant decrease in blood pressure could be observed (-7.3/-4.0 mmHg), which was all the more pronounced, the higher the baseline values had been. CONCLUSION: Under real practice conditions, for overweight and obese patients treatment with sibutramine led to significant weight reductions and was well tolerated. The metabolic and cardiovascular risk profile was significantly improved, which was especially beneficial for patients with a high baseline risk. PMID- 11907863 TI - [Intestinal pseudoobstructions and gastric necrosis in mitochondrial myopathy]. AB - HISTORY AND CLINICAL FINDINGS: A 24-year-old female patient suffered for 4 months from recurrent abdominal pain, vomiting and diarrhea. Signs of an acute abdomen were the initial reason for admitting the patient to our hospital. The slim, pale patient had a complete bloated abdomen. Neurological status was normal. INVESTIGATIONS, TREATMENT AND COURSE: Radiographic examination showed a paralytic ileus with a megacolon. The recurrent abdominal symptoms were due to a covered perforation of the stomach. This was shrunken, scarred and had to be resected. Further intestinal pseudoobstructions were accompanied by substantial exsudations in the lungs, intestines and abdomen. At this time mutism like behavior patterns and an ophthalmoplegia appeared. Leukoencephalopathy in brain MRI scans and increased liquor-lactate suggested mitochondrial myopathy. DIAGNOSIS: The diagnosis of a mitochondrial myopathy was confirmed by increased liquor-lactate level, muscle biopsy with ragged-red fibers as well as abnormal mitochondrias and molecular-genetic investigations (mtDNA point mutation A3243G). Associations to MELAS (mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes) and MNGIE (mitochondrial neuro-gastrointestinal encephalomyopathy) syndrome are discussed. CONCLUSIONS: Unclear recurrent gastrointestinal symptoms even in the absence of neurological changes may reflect a mitochondrial disease. This applies especially to young patients with recurrent anorexia, vomiting and pseudoobstruction. In case of additional symptoms like ophthalmopathy, deafness, diabetes mellitus or signs of a MELAS syndrome the search for a mitochondrial system disorder is mandatory. PMID- 11907864 TI - [Pyrethroid exposure following indoor treatments with a dog flea powder]. AB - Pyrethroid exposure following indoor treatments with a dog flea powder. HISTORY: A 42 year old woman reported hair loss, gastrointestinal and non-specific symptoms. The patient has lived in a council flat and kept a dog who had been regularly treated with pyrethroid containing flea powder. INVESTIGATIONS: The biological monitoring of pyrethroid meta-bolites in urine was performed using gas chromatography-mass spectrometry. The values at admission and follow-up after 4 weeks were highly elevated. Inspection of the flat revealed a humid and cramped dwelling. TREATMENT: We recommended redevelopment and cleaning of the dwelling and the avoidance of ectoparasiticide use. CONCLUSION: To our knowledge this is the first documented case of high indoor pyrethroid exposure following the use of ectoparasiticides with domestic animals. Pyrethroids can cause neurotoxic symptoms and skin irritation. There are few data concerning chronic effects. The causal connection between pyrethroid exposure and symptoms remains unclear and poses a great problem in environmental medicine. PMID- 11907865 TI - [Stem cells]. PMID- 11907866 TI - [Therapeutical use of parathyroid hormone]. PMID- 11907868 TI - [Informed consent: ethical and legal aspects]. PMID- 11907867 TI - [Framework for biomedical research involving human subjects - algorithms for planning and organisation]. PMID- 11907869 TI - [Bisphosphonate therapy in Paget disease?]. PMID- 11907870 TI - New Zealand health reforms: effect on ophthalmic practice. AB - Are specialized ophthalmic units with inpatient facilities going to disappear in the New Zealand public health system? We have entered the era of cost containment, business methodologies, bench marking, day case surgery, and technologic advances. The dilemma for nursing is maintenance of a skill base with dwindling clinical practice areas. PMID- 11907871 TI - Preparing for the CRNO examination: it's time to get started. AB - The focus of this article is to suggest ways in which a nurse can prepare for the certification examination for ophthalmic registered nurses (CRNO). PMID- 11907872 TI - Hands across the patient: primary team nursing in an ambulatory surgicenter. PMID- 11907873 TI - Complications of extraocular muscle surgery. AB - This article reviews the goals, complications, interventions, and potential outcomes of extraocular muscle surgery. The information will support nursing interventions in the operating room and ambulatory care arenas, as well as those in school or public health settings. PMID- 11907874 TI - Get some culture: the need for cross-cultural awareness is closer than you think. PMID- 11907875 TI - Minor surgery: an extended role for ophthalmic nurses in the United Kingdom. AB - This article discusses the reasons for extending the scope of ophthalmic nursing practice in the Princess Alexandra Eye Pavilion, Edinburgh, Scotland, particularly with regard to the ophthalmic nurses' role and management of meibomian cysts (chalazions) through minor surgery to the eyelid. The article describes how the in-service teaching program was established in the hospital for those ophthalmic nurses who were interested in developing their knowledge and practical surgical nursing skills to undertake the management of meibomian cysts. The appropriate legal and ethical aspects of patient care involved in extending the scope of practice in the United Kingdom are discussed, including accountability and duty of care. PMID- 11907876 TI - Look for green grass--not poop--and you can refresh your job attitude! PMID- 11907877 TI - Continuous quality improvement analysis of operating room turnover time. AB - The continuous quality improvement process reminds us how recognizing opportunity for growth enhances the provision of quality patient care. PMID- 11907878 TI - The nurse practitioner role in a United Kingdom ophthalmic accident and emergency department--10 years of progress. AB - This article considers a unique, nurse practitioner (NP)-led accident and emergency service within a large ophthalmic hospital in the United Kingdom. The development of this service from a traditional, ophthalmologist-led department is traced, along with the original intent of the NP role and how it has evolved. Positive and negative aspects of the NP role and the results of the service's first audit are discussed. The service has proved very successful, and the NP role has been and will continue to be dynamic, with NPs totally committed to delivering the best possible care for their patients. PMID- 11907879 TI - Choroidal metastasis from prostate adenocarcinoma. PMID- 11907880 TI - Implementation of an innovation in the ophthalmic operating room: the Memory lens. AB - I have participated in the evolution of the cataract procedure for the last 23 years. This year, as nursing director for an eye surgery center, I was responsible for the implementation of a new prerolled intraocular lens: The Memory lens. I review 5 implementation strategies, which are: Be open to recently approved products Utilize all company resources Introduce, train, train, train Evaluate, review, review, review Enjoy the questions and attention from others considering the innovation The implementation of this innovation confirmed both my and the operating room staff's ability to change and reaffirmed their commitment to achieve the resultant patient benefits. PMID- 11907881 TI - Nonproliferative diabetic retinopathy and macular edema. AB - As previously noted, although visual loss usually does not fall below 20/200 in the presence of ME, it may nevertheless be a significant disability. Additional interventions may include referral to low vision clinics, home health agencies, visual loss support groups, and local or regional blindness agencies to aid the patient's occupational rehabilitation, coping mechanisms, and adaptation responses in the presence of this potentially debilitating process. Control of blood sugar, blood pressure, and the intervention of focal/grid laser treatments to seal leaks and prevent further edema provide the best chance of maintaining useful vision throughout life. Patient education is paramount to improve comprehension of the condition, recommended treatment modalities, and compliance with prescribed regimens. Assessments and interventions related to knowledge and sensory deficits, anxiety, discomfort, ineffective coping mechanisms, and health maintenance behaviors add a quality link in the multidisciplinary approach surrounding the delivery of care to patients with NPDR and clinically significant ME. PMID- 11907882 TI - To have or not to have--mutual recognition for nursing practice. PMID- 11907883 TI - Impact of glaucoma. Part 1: Burden of the disease. PMID- 11907884 TI - Childhood glaucoma. PMID- 11907885 TI - Glaucoma: what the ophthalmic nurse should know. AB - Glaucoma is both an international and a national public health issue. Worldwide, glaucoma is one of the leading causes of blindness. More than 2 million persons in the United States have glaucoma, yet only half of them are aware that they have the disease. The diagnosis of glaucoma requires an extensive ocular examination. Glaucoma consists of a group of ocular diseases that result in optic disk cupping and visual field loss. Although glaucoma is a blinding disease, in most cases, blindness may be prevented through early detection and treatment. Glaucoma management is determined by the type of glaucoma a person has. PMID- 11907886 TI - Hot items: certification examination questions. AB - This article discusses the conception and development of test items for the certification examination for ophthalmic registered nurses. PMID- 11907887 TI - Sight-saving mission changes personal perspective. PMID- 11907889 TI - Annular tinted contact lenses caused irregular corneal astigmatism. PMID- 11907888 TI - Endophthalmitis. AB - Endophthalmitis may change a person's outlook on life completely, leading him or her to wonder what went wrong and why he or she is not seeing. PMID- 11907890 TI - The geriatric depression scale (GDS). PMID- 11907891 TI - Modeling interpersonal competencies enhances patient care and the organization. PMID- 11907893 TI - Enucleation: indications, methods, and prosthetic options. PMID- 11907892 TI - Non steroidal anti-inflammatory aents. PMID- 11907894 TI - Corneal abrasion: to patch or not to patch. PMID- 11907895 TI - Analyzing the quality of ophthalmic nursing research. AB - The purpose of this project was to describe, classify, and analyze ophthalmic nursing research published between 1975 and 1996. Sixty-six quantitative research articles were reviewed. This mainly descriptive body of research covered such topics as assessment/screening, self-care, and risk factors. The research articles were generally rated as superior or average, with many of the average being at the low end of the scale. Twenty-eight percent of the research was not applicable to nursing practice, and 42% had low or no theoretical relevance to nursing. This review will help to establish a research agenda in the area of ophthalmic nursing. PMID- 11907896 TI - First do no harm: routine use of aminoglycosides in the operating room. AB - For decades aminoglycosides have been used routinely in intraocular surgery to prevent and treat endophthalmitis. However, as more and more studies are conducted, the reality is that these drugs are causing more harm than they are preventing. Summarized herein are the most recent studies concluding that even low-dose aminoglycosides can cause toxic retinal damage. The conclusion, with pertinent recommendations from the Johns Hopkins Hospital, Wilmer Eye Institute Infection Control Committee, is that the questionable benefit of use of aminoglycosides intraoperatively is not justified by the risk of injury. PMID- 11907897 TI - Developing the role of the ophthalmic nurse in England. AB - The role of the ophthalmic nurse in England has developed during the last 5 years with more nurses working as practitioners who run their own clinics. With the introduction of Working for Patients, the whole ethos of the National Health Service in England has changed. This study uses structured interviews to identify how three Trusts have developed the role of the ophthalmic nurse to meet the changing needs of the National Health Service. Analysis of the data identified that despite numerous problems encountered, ophthalmic nurses are responsible for driving same-day surgery ensuring that patients' needs are met. PMID- 11907898 TI - No end in sight: the nurse's role in ensuring a holistic approach in the care of a patient undergoing a drainage implant for end-stage glaucoma. AB - This article addresses issues that relate to the surgical care of patients in imminent danger of permanently losing their eyesight as a result of advanced glaucoma. An overview of the stages of glaucoma and concurrent treatment is reviewed. Close attention is given to the psychosocial and physiologic ramifications of this end-stage surgical procedure. The nurse's role in assisting patients through this life crisis is essential to the patient's well being, lifestyle changes, and quality of life. PMID- 11907899 TI - Complications of punctal plugs. PMID- 11907900 TI - The Pittsburgh Sleep Quality Index (PSQI). PMID- 11907901 TI - Introduction to oculoplastic surgery. PMID- 11907902 TI - Unusual presentations of dentigerous cysts. PMID- 11907903 TI - Post-exposure treatment & cost containment tips. PMID- 11907904 TI - Time management and feelings of accomplishment are directly related! What is wrong with this statement? PMID- 11907905 TI - A study: will waiting room time be reduced when the redesigned preliminary workup is performed by a certified ophthalmic technician? AB - The primary source of patient dissatisfaction in ophthalmic clinics is waiting room time. At one clinic, a study was conducted to determine whether redesignating preliminary workup tasks affected waiting room time. Previously, technicians and the ophthalmic registered nurse performed workups by taking histories, checking vision, reading glasses, performing refractions, conducting tonometry and slit lamp examinations, and dilating patients' eyes. Because the technicians' refractions were often inaccurate, the nurse or the doctor had to redo the refraction, prolonging waiting room times. Workup tasks, beginning with refractions, were transferred from the technicians to the nurse, which increased patient satisfaction and staff efficiency. PMID- 11907906 TI - Silicone oil-induced secondary glaucoma: a case study. AB - Silicone oil intraocular tamponade is a widely accepted procedure in the management of complex retinal detachments caused by proliferative diabetic vitreoretinopathy. Silicone oil has a high surface tension that mechanically limits fibrovascular reproliferation resulting in successful retinal reattachments. However, postoperative secondary glaucoma is a relatively frequent complication that may require intensive nursing management focused at intraocular pressure monitoring, positioning compliance, pain management, fluid and electrolyte balance, and glucose control. This article presents the postoperative clinical course of a 51-year-old man with diabetes and recurrent proliferative diabetic vitreoretinopathy who developed secondary glaucoma after silicone oil injection with a dramatic rise in intraocular pressure on the first postoperative day. Nursing management concurrent with medical and surgical management is discussed and the necessary nursing plan of care is identified. PMID- 11907907 TI - How a multidisciplinary team improved a process to attain positive patient outcome. AB - A case study approach will be used to demonstrate the effectiveness of a continuous quality improvement team designed to explore the process of preoperative medical record completeness. The goal of this team was to improve essential chart components (e.g., electrocardiography, history and physical examination) required 48 hours before the date of surgery in an effort to facilitate optimum patient management and timely flow of patients to the operating room. When the 48-hour compliance rule is followed, the nurse reviewer has sufficient time to review all patient data and notify attending physicians or designees of any abnormalities. In essence, improving component compliance provides better quality care because patients are able to be assessed before surgery with enough time to solve any problems. PMID- 11907908 TI - Volunteering at an eye camp: fulfillment in the heart of India. PMID- 11907909 TI - Ocular injuries induced by lightning. PMID- 11907910 TI - Prevent Blindness America: our local chapter experience. PMID- 11907911 TI - Ocular toxoplasmosis. PMID- 11907912 TI - Implementing a nurse-led telephone advice system in ophthalmology. AB - This article describes the background research I undertook to determine the potential for a telephone advice line in this major specialist eye hospital in the United Kingdom. The information gained was used to implement a computer-based telephone assessment system with guidelines. This system enables a registered nurse with an ophthalmic qualification to offer telephone triage and general information and advice about eye problems. The help line was identified as a service that would enhance the care of patients with eye problems, their carers, and health providers. The help line, "Moorfields Direct," was given a dedicated telephone number and was officially launched in February 1999. PMID- 11907913 TI - Eye banking: the business of restoring sight. AB - This continuing education article provides an overview of how an eye bank operates, offers general criteria about who can be a donor, and introduces the process by which eye donation takes place. In presenting this information, we hope to bring honor to the memory of all eye donors and thank them, their loved ones and families for this truly philanthropic gift. PMID- 11907914 TI - A systematic review of the skeletal effects of estrogen therapy in postmenopausal women. II. An assessment of treatment effects. AB - PURPOSE: To combine the results of randomized controlled trials to provide overall estimates of the effect of estrogen treatment on fracture rates and measures of bone mass. DATA SOURCES: Articles on estrogen treatment for osteoporosis published between 1977 and 1995 were identified. STUDY SELECTION: Studies selected were randomized controlled trials of the efficacy of estrogens in preventing loss of bone mass or fractures in postmenopausal women. DATA EXTRACTION: Data extraction and quality assessment were performed in duplicate, with assistance of a manual. Raters were blinded as to authors and their affiliations and the publication details. With estimates of bone mass, the treatment effect size was defined as the difference in the mean annual change in bone mass between the treatment and control groups divided by the pooled standard deviation for change. In the case of fractures, efficacy was measured as the reduction in the numbers of individuals experiencing new fractures with treatment. Effect sizes were pooled using the random effects model. RESULTS: Thirty-seven studies met the criteria for inclusion in the systematic review. Only one small secondary prevention trial contained evaluable data on vertebral fractures. This study found a fracture relative risk of 0.63 (95% confidence interval, CI 0.28-1.43) with estrogen treatment. There was more information on the effects of treatment on bone mass. Overall effect sizes ranged between 0.5 and 2.5 standard deviation (SD) units for change. A dose-response relationship was apparent but high doses of estrogens were not associated with effect sizes greater than those observed with recommended doses. There was no significant difference in efficacy between transdermal and oral administration of estrogens. Pooling of paired data from secondary prevention studies indicated that treatment effect sizes were smaller at the hip (0.92, 95% CI 0.3-1.5 SD units) than at the spine (2.1, 95% CI 0.9-3.3 SD units). No significant effects of co-intervention with calcium, progestogens or androgens were seen, although an additive effect of higher doses of calcium could not be ruled out. CONCLUSIONS: Clear-cut effects of estrogens in attenuating the postmenopausal decline in estimated bone mass were apparent in this literature. However, the trials were short-term and provide inadequate evidence on the effects of treatment on fracture risk. PMID- 11907915 TI - Effects of dietary phytoestrogens in postmenopausal women. AB - The aim of this study was to test the hypothesis that increased dietary intake of phytoestrogens reduces the health impact of the menopause. To test this hypothesis, a double-blind, randomized, entry-exit, cross-over study was conducted to assess the effects of three dietary manipulations--soy and linseed diets (high in phytoestrogens) and a wheat diet (low in phytoestrogens). Postmenopausal women were recruited and randomly assigned to one of the three dietary regimens. Urinary phytoestrogen concentrations, hot flush rate, vaginal smears, bone mineral density and bone mineral content were assessed for two 12 week periods. Comparative analysis showed no significant differences, but, when analyzed separately, groups consuming high phytoestrogen diets had between 10 and 30 times higher urinary excretion of phytoestrogens compared to those consuming the low phytoestrogen diet (p < 0.01). Study participants consuming soy, linseed and wheat diets had a 22% (not significant, n.s.), 41% (p < 0.009) and 51% (p < 0.001) reduction in hot flush rate; a 103% (p < 0.04), 5.5% (n.s.) and 11% (n.s.) increase in vaginal cytology maturation index; and a 5.2% (p < 0.04), 5.2% (n.s.) and 3.8% (n.s.) increase in bone mineral content, respectively. No changes were detected in bone mineral density. The differential effects of high phytoestrogen dietary manipulations on outcomes may represent tissue-specific responses to isoflavones and lignans contained in soy and linseed, respectively. Whilst health outcome measures were not significantly different between groups, the data obtained from separate analysis suggest that phytoestrogens in soy and linseed may be of use in ameliorating some of the symptoms of menopause. Furthermore, the significant decrease in hot flush rate in the wheat group cannot be attributable to phytoestrogens measured in this study. Due to subject variability, larger studies are still needed to evaluate population benefit. PMID- 11907916 TI - A cohort study of hormone replacement therapy given to women previously treated for breast cancer. AB - Women who have been previously treated for breast cancer are usually advised to avoid hormone therapy for fear of increasing their risk of tumor recurrence. However, for some women, menopausal symptoms are so severe that their quality of life is poor. Because ethic committees are reticent to permit a double-blind randomized trial, we performed a cohort study of hormone therapy after breast cancer. METHODS: The study group comprised 1472 women with breast cancer. A total of 167 subjects had used an oral or transdermal estrogen after their treatment for breast cancer. Amongst these estrogen users, 152 (91%) had also used a progestin. In total, 106 other women had used a progestin alone as a treatment for menopausal flushes and not as a treatment for breast cancer. Cox regression analysis was performed using estrogen as a time-dependent covariate with disease free interval as the outcome. RESULTS: The uncorrected hazard ratio for the estrogen-progestin users was 0.67 (95% confidence interval (CI) 0.38-1.16) and for the progestin alone users was 0.85 (95% CI 0.44-1.65). CONCLUSIONS: This study was unable to demonstrate a significant increase in risk of breast cancer recurrence for women who used HRT and suggests that the time is now appropriate for a randomized prospective trial of hormone therapy after breast cancer. PMID- 11907917 TI - Selective estrogen receptor modulators. PMID- 11907918 TI - Treatment of estrogen deficiency symptoms in women surviving breast cancer. PMID- 11907919 TI - Cancer, cohorts, confounding and consensus. PMID- 11907920 TI - Hormone replacement therapy and the skeleton: what does the evidence tell us? PMID- 11907921 TI - A systematic review of the skeletal effects of estrogen therapy in postmenopausal women. I. An assessment of the quality of randomized trials published between 1977 and 1995. AB - PURPOSE: To examine the quality of published randomized controlled trials of the effects of estrogen treatment on fracture risk and measures of bone mass. DATA SOURCES: Articles on estrogen treatment for osteoporosis published between 1977 and 1995 were identified by searching Medline and Excerpta Medica databases and bibliographies of original papers and published reviews. STUDY SELECTION: Studies selected were randomized controlled trials of the efficacy of estrogens in preventing loss of bone mass or fractures in postmenopausal women. DATA EXTRACTION: Data extraction and quality assessment were performed in duplicate, with assistance of a manual. Raters were blinded as to authors and their affiliations and the publication details. RESULTS: Of 99 eligible randomized controlled trials published between 1977 and 1995, eight included no extractable data, and 23 contained results that were published in duplicate. Total quality scores increased over time, but this was accounted for by improvements only in the measurement technologies used to estimate bone mineral content or density. There was no improvement in the quality of randomization methods, the extent to which withdrawals were accounted for, or in the baseline comparability of treated and control patients. Neither sample sizes nor durations of follow-up increased over time. CONCLUSIONS: This body of literature fails to address whether estrogen therapy reduces fracture rates, and does not allow for comparison of the effects of different active therapies on change in bone density. Although there were improvements in the techniques for estimating bone mass and delivering estrogen treatment, the studies published in the 1990s were no more informative for making clinical or policy decisions than those published in the 1970s. PMID- 11907923 TI - The change to Climacteric. PMID- 11907922 TI - Using longitudinal data to define the perimenopause by menstrual cycle characteristics. AB - OBJECTIVES: To determine which aspects of menstrual change best predict time to postmenopause. METHODS: A total of 250 Australian-born women aged 45-55 years were divided into five menstrual status categories: Group I reported no change in menstrual flow or frequency; Group II reported change in flow; Group III reported change in frequency; Group IV reported change in both frequency and flow; and Group V reported between 3 and 11 months of amenorrhea. Menstrual status groups were compared on baseline data for age, hormone levels, hot flushes and self rated menopausal status. The proportion of women moving to postmenopause in subsequent years was compared using 4 years of follow-up data. RESULTS: Women in Group V were older, had lower estradiol and inhibin levels, higher follicle stimulating hormone levels, and were more likely to report hot flushes, and to self-rate themselves as having started the menopausal transition, compared with the women who had menstruated in the last 3 months (Groups I-IV). Groups I and II were similar in age and hormonal status, as were Groups III and IV. The proportion of women who had moved to postmenopausal status in the 4 years after baseline were 12%, 14%, 58%, 53% and 94% for Groups I-V, respectively. CONCLUSIONS: Amenorrhea is the best predictor of future menopause followed by changes in menstrual frequency. Change in flow only was not predictive of future menopause. A two-stage classification scheme is suggested for defining the perimenopause. 'Early perimenopause' is defined as the self-reporting of changes in menstrual frequency over the last year, and 'late perimenopause' is defined as the self-report of 3-11 months of amenorrhea. PMID- 11907924 TI - Acute effects of a surgical menopause on serum concentrations of lipoprotein(a). AB - OBJECTIVE: The aim of this study was to investigate acute changes in serum concentrations of lipoprotein(a) and other atherogenic lipids and lipoproteins after a surgical menopause. METHODS: A total of 100 premenopausal Chinese women who were booked for hysterectomy for benign gynecological disorders were recruited. They study group comprised 40 subjects undergoing hysterectomy as well as bilateral oophorectomy. The control group consisted of 60 subjects undergoing hysterectomy with conservation of the ovaries. Complete data were available from 30 of the 40 subjects in the study group and from 44 of the 60 controls. Serum concentrations of lipoprotein(a) and other atherogenic lipids and lipoproteins were measured before surgery and these measurements were repeated 3 days, 8 weeks and 6 months postoperatively. Those study patients who received hormone replacement therapy and control patients who became menopausal, according to biochemical criteria, during the study period were excluded from analysis. RESULTS: Three days after surgery, there was a significant increase in the mean lipoprotein(a) concentration in the control group from 19.1 to 23.0 mg/dl (p < 0.01), but there was no significant change in the study group. There were no significant changes from baseline in the mean lipoprotein(a) concentration in either group 8 weeks or 6 months after surgery. There was a significant increase in the mean concentration of total cholesterol in the study group 8 weeks after surgery from 5.08 to 5.45 mmol/l (p < 0.01), in low density lipoprotein cholesterol from 3.22 to 3.49 mmol/l (p < 0.01), and in apolipoprotein B from 95.6 to 103.0 mg/dl (p < 0.05). However, the mean concentrations 6 months after surgery were not significantly different from baseline levels. The mean concentrations of high density lipoprotein cholesterol, apolipoprotein A-1 and triglycerides also did not differ significantly from baseline in the study group, either 8 weeks or 6 months after surgery. CONCLUSIONS: These results suggest that any increase in concentrations of lipoprotein(a), and other atherogenic lipids and lipoproteins which occur after the menopause, develops relatively slowly. The changes in concentrations which occurred within 8 weeks of surgery were probably an acute-phase reaction after surgery rather than a response to a decreasing estradiol concentration. PMID- 11907926 TI - Where is the evidence for effectiveness of treatments for the menopause? Introducing the Cochrane Collaboration. AB - The past two decades have seen an explosion of research surrounding the menopause. This ever-increasing volume of research has made it an impossible task for health professionals involved in the management of the menopause to be able to assimilate the information easily. There is an urgent need for the findings from research to be synthesized into simple, easy-to-read reviews that are of high quality and are based on the best evidence available. The Menstrual Disorders and Subfertility Group of the Cochrane Collaboration is attempting to address these issues by collecting a register of all the randomized controlled trials in the treatments of the menopause and preparing systematic reviews on a number of topics that will be of interest to health-care workers and consumers. Readers are invited to participate in this process by identifying published and unpublished data and by helping in the process of preparing protocols and systematic reviews for inclusion in the Cochrane Library. PMID- 11907925 TI - Hormone replacement therapy: prevalence, compliance and the 'healthy women' notion. AB - BACKGROUND: The objective of this study was to assess the current trends of hormone replacement therapy (HRT) use, including rates of use, length of use, continuation rates and characteristics of users and non-users and to examine the hypothesis that 'healthy women' are more likely to be users of HRT. METHODS: Analysis was carried out of three representative South Australian population studies in 1991, 1993 and 1995 comprising 3019, 3004 and 3016 personal interviews, respectively. RESULTS: Current use (and ever-use) of HRT in all women aged 50 years and over rose from 13.2% (26.7%) in 1991, to 21.2% (31.9%) in 1993 and 26.0% (40.5%) in 1995. Highest use is now in the 55-59-year age group where, in 1995, current use was 50.9% and ever use was 69.0%. Median compliance rates with HRT rose from 24 months in 1991 to 60 months in 1995 for current users aged 50 years of age or above. The pattern of increasing use of HRT is not consistent across age groups. Analyses of the 1995 data show that, in contrast with increasing rates of current use in women over 55 years, there was no overall change in rates for women below this age. There were no statistically significant differences in health indicators, e.g. blood pressure, smoking, cholesterol levels or body mass index between users and non-users of HRT. However, users reported significantly higher rates of previous osteoporosis and hysterectomy. CONCLUSIONS: Prevalence rates of HRT use are increasing together with compliance rates and this may reflect increased confidence with HRT therapy. Users of HRT have an increased rate of mammography compared to non-users and this may contribute to earlier detection and, therefore, increased estimates of breast cancer in HRT users. There was no support from the 1995 data for a 'healthy women' hypothesis among HRT users. PMID- 11907927 TI - Health status of hormone replacement therapy users and non-users as determined by the SF-36 quality-of-life dimension. AB - BACKGROUND: The objective of this study was to compare the health status of women who use and do not use hormone replacement therapy (HRT). METHOD: The 1994 South Australian Health Omnibus Survey (a population health interview survey) was used to administer the short form-36 health survey questionnaire (SF-36) to users and non-users of HRT. A representative sample of 813 women aged 40 years and older were interviewed. The response rate of the survey was 72.4%. Eight health dimensions of the SF-36 were measured: physical functioning, social functioning, role limitations owing to emotional problems, role limitations owing to physical problems, mental health, vitality, pain and general health. RESULTS: The mean score for all eight health dimensions was in the bottom 50% of the population for HRT users while non-users were in the upper 50%. Users of HRT had significantly poorer scores for physical limitations, body pain, general health, vitality, social functioning and mental health. CONCLUSION: Women who use HRT are less healthy than non-users when measured by a generic health status measure. PMID- 11907928 TI - Hormonal treatment and psychological function during the menopausal transition: an evaluation of the effects of conjugated estrogens/cyclic medroxyprogesterone acetate. AB - OBJECTIVE: To investigate the effect of hormone treatment on psychosocial distress of women during the menopausal transition (aged 40-52 years, still menstruating and with minor symptoms), in a study carefully designed to reduce bias and placebo effect. METHODS: The study was randomized, controlled by placebo, blinded to the subject, investigators and biostatistician, crossed-over after 6 months, and evaluated by a 77-item questionnaire every month for 12 months. The medication tested was continuous conjugated estrogens (Premarin) and cyclic medroxyprogesterone acetate (Provera). RESULTS: Of an eligible 105 apparently healthy women, recruited by advertisement, 83 completed the questionnaires for the study. Randomization was successful. While there was a substantial variation over time in the change of scores for all the psychosocial outcomes, there was no significant difference between active and placebo treatment when order of treatment allocation was ignored. However, a strong and consistent effect of the order of treatment allocation for many of the scores was found; in particular, the effect of active treatment was substantially stronger when it was administered second. Positive effects of active treatment were found for the score for the overall symptom rating test (p < 0.009) and its components of depression and feeling of inadequacy (p = 0.011; p = 0.001, respectively). CONCLUSIONS: The use of hormones, given as a continuous estrogen and cyclic progestogen formulation after a formal calibration/run-in period, may have a beneficial effect on psychosocial distress experienced by women towards the end of their reproductive function. PMID- 11907929 TI - Microanatomy of the female reproductive organs in postmenopause by scanning electron microscopy. AB - The detailed three-dimensional ultrastructural features of the reproductive organs of menopausal and postmenopausal women were studied by means of integrated transmission and scanning electron microscopy (SEM) and reported in a new colored microtopographical fashion. These methods revealed significant alterations in the microanatomy of the various reproductive organs specifically related to the decline of plasma estrogen levels. In particular, the ovary progressively showed characteristic wide areas of loss of epithelium with consequent exposure of the underlying connective tissue. Both endometrial and tubal mucosa demonstrated a gradual but often dramatic decrease in the number of ciliated cells which was more evident in the tube. In addition, the non-ciliated (microvillous secretory) cells of the uterus, including both endocervix and tubal mucosa, became flattened and, in some instances, their apical poles developed unusual wrinkles (microridges or microplicae). The ectocervix and vaginal squamous cells presented a reduction in the number of their microridges and changes in the typical structural organization. These microtopographical results showed that the decline of estrogen during the menopause and postmenopause induces important and complex structural changes of the woman's reproductive system, much more detailed than those revealed to date by the use of only conventional optical and transmission electron microscopy (TEM). The three-dimensional findings offer the opportunity to re-evaluate the classic histopathology of the above aging organs using more refined microtopographical and morphophysiopathological parameters. PMID- 11907930 TI - Postmenopausal bone remodelling and hormone replacement. AB - Osteoporosis is a serious, frequently occurring disease. Sensitive, specific and precise biochemical measures of bone remodelling, thus, are important tools in the evaluation of bone loss, fracture risk and treatment response. Bone turnover is predictive of the subsequent rate of loss. A high bone turnover documented by increased levels of biochemical markers predicts an increased rate of loss in bone mass. Additionally, as a high bone turnover leads to an increase in the extent of bone resorption as well as an increase of the erosion depth, this may also cause deterioration of the bone structure, thereby leading to an increased risk of fracture. That the predictive values of bone mass and markers of bone turnover are additive suggests that these measures describe different properties of bone. Hormone replacement induces a reduction in postmenopausal bone turnover, arrests loss of bone mass and decreases fracture risk. The skeletal response to therapy is reflected by bone markers and these may be used for monitoring purposes. Those who are in most need of treatment can be identified by a combination of bone mass and bone marker measurement. Furthermore, those who need the treatment most will demonstrate the best response in terms of bone mass. Concern has arisen because of a relatively large day-to-day variation, especially in urinary markers; however, this seems to be resolved through use of the new serum markers. PMID- 11907931 TI - Electrocardiogram pattern in hypercholesterolemic women: the influence of hormone replacement therapy. AB - OBJECTIVE: The aim of this study was to delineate electrocardiogram (ECG) patterns in postmenopausal women with hypercholesterolemia and to assess the possible influence of female sex hormones. STUDY DESIGN: A total of 72 postmenopausal women with moderately elevated total cholesterol levels constituted the case group, of which 48 came from a clinical trial and 24 from a cohort study. Some 236 women aged 50-59 years with normal levels of cholesterol were participants in the same cohort study in the local area. These 236 women had been subdivided into three groups: premenopausal, postmenopausal and postmenopausal with hormone replacement therapy (HRT). Of the 48 women in the clinical study group, 12 patients showed pathological ECG changes. Six of these patients were treated with HRT for 2 years (transdermal estradiol 50 micrograms/day and a daily dose of 5 mg medroxyprogesterone acetate, MPA) and the rest were non-users of HRT. RESULTS: In the women with hypercholesterolemia, 16 of 72 patients (22%) showed ischemic ECG changes, compared to nine of 88 (10%) with normal cholesterol levels (p = 0.04). We found no significant difference in the prevalence of ECG changes between postmenopausal women with and without HRT in the groups with normal levels of cholesterol. In the hormone treatment group, four of six patients showed an improvement in ECG pattern, in contrast to two of six non-users of HRT. CONCLUSIONS: This preliminary study revealed a higher prevalence of pathological ECG changes in postmenopausal women who had hypercholesterolemia than in normocholesterolemic women. These findings support the idea that hyperlipidemia contributes to the overall increase in cardiovascular disease, as this is also associated with ECG changes. Transdermal estradiol combined with MPA has a beneficial effect in reversing the process of atherosclerosis, as well as improving the ECG pattern. The prevalence of pathological ECG patterns was similar for HRT users and non-users. This outcome may be affected by several factors. Hence, further research is warranted. PMID- 11907932 TI - Mental health among perimenopausal women attending a menopause clinic: possible association with premenstrual syndrome? AB - BACKGROUND: Psychiatric symptoms are frequent in the perimenopause. They are similar to symptoms observed at different stages of a woman's life cycle, suggesting that there may be an association between mental disturbances of the perimenopause and those observed during premenstrual and postnatal periods. METHODS: This study aimed to determine the reliability of using a modified version of the Steiner premenstrual tension self-rating scale (PMTS) questionnaire for assessing retrospectively the presence of premenstrual complaints and to evaluate the association between previous premenstrual complaints and psychiatric symptoms at the time of the menopause. Forty-one perimenopausal women were selected to establish the reliability of the questionnaire to assess premenstrual symptoms retrospectively (4-8-week interval between measures); agreement between measurements was assessed using the kappa statistic. Ninety-six women were later recruited from a gynecological menopause outpatient clinic to study the association between premenstrual complaints and the presence of psychiatric symptoms at the time of the menopause (as measured by another self-reporting questionnaire, the SRQ-20); SRQ-20 total scores greater than 7 were considered to be indicative of psychiatric morbidity. RESULTS: All 36 PMTS items showed moderate to very good test-retest reliability (0.44 < kappa < 1.0). There was a significant correlation between total PMTS and SRQ-20 scores (Spearman correlation coefficient 0.75, p < 0.001), correlation coefficient and SRQ-20 total scores greater than 7 were found in 47.9% of patients. CONCLUSIONS: Premenstrual symptoms can be reliably measured in perimenopausal women. Women who report having experienced premenstrual dysphoria are more likely to present with psychiatric symptoms at the time of the menopause. PMID- 11907933 TI - Relationship between menopausal stage and age and quality of relationships with partners, children and friends. AB - OBJECTIVES: This study investigated the interaction of menopausal stage and age with the quality of women's relationships with their partners, children and close friends. METHODS: Females from a community sample (n = 48, age range 35-65 years) volunteered to participate in structured interviews. Questions relating to demographic background and relationships with partners, children and friends were included. Seventeen women were premenopausal, 16 women had been postmenopausal for 1-5 years and 15 women had been postmenopausal for between 6 and 10 years. RESULTS: The association of age and menopausal stage could not be separated when relationships with partners were considered. Premenopausal and younger women appeared to be more satisfied and more positive, and rated their relationship as more important to them than did older and postmenopausal women. Menopausal stage, but not age, appeared to be associated with the satisfaction women had with their children and the desire to see their children more often. When the two groups of postmenopausal women were compared, it was found that those women postmenopausal for 1-5 years wanted to see their children more often and were more dissatisfied with the relationship they had with their children than those women postmenopausal for 6-10 years. There were no effects of menopausal stage or age when the quality of close friendships was considered. CONCLUSIONS: Further work is needed to explore the implications of these findings as they relate to the interaction of menopausal stage and age with the quality of interpersonal relationships. PMID- 11907934 TI - Asymptomatic postmenopausal intrauterine fluid accumulation: characterization and significance. AB - OBJECTIVES: The aims of the study were to characterize those postmenopausal women who develop intrauterine fluid accumulation and to evaluate its significance. METHODS: All asymptomatic postmenopausal women who were referred for routine transvaginal ultrasonographic examination between 1 January 1995 and 31 March 1996 were included in the study. Demographic and ultrasonographic parameters were recorded on a prospectively created computerized database. When intrauterine fluid accumulation was identified, the women was referred for endometrial sampling. RESULTS: A total of 1175 consecutive, asymptomatic postmenopausal women were evaluated; intrauterine fluid accumulation was identified sonographically in 166 (14.1%). Women with intrauterine fluid accumulation were older, had experienced more years since the menopause, and had smaller uterine volume indices, thinner endometria and smaller indices of ovarian area, compared to those without intrauterine fluid accumulation (all at a significant level of p < 0.0005). The prevalences of hormone replacement therapy use were 6.6% in the 'accumulating fluid' women and 43% in the 'non-accumulating fluid' group (p < 0.0005). Of the 166 women with intrauterine fluid accumulation, 91 had an endometrial biopsy, of which 70% were insufficient for evaluation and 30% were normal on histology. CONCLUSION: Postmenopausal intrauterine fluid accumulation is a common, mostly benign phenomenon that typically occurs in the late postmenopausal age subgroups. It may be postulated that it represents part of the atrophic mechanism that takes place at this stage of life. Hormone replacement therapy appears to be a 'protection' against this phenomenon. PMID- 11907935 TI - Pharmacokinetic comparison between Conpremin (Premarin) and a generic preparation of conjugated estrogens. AB - OBJECTIVES: To determine the relative bioavailability of the estrogenic components of a generic brand of conjugated estrogens marketed in Chile in comparison to that of Conpremin (Premarin in the United States). METHODS: A randomized cross-over study was conducted on 16 healthy postmenopausal women receiving single oral doses of either two Conpremin 0.625-mg tablets or two 0.625 mg tablets of the generic brand, with a 14-day wash-out interval between doses. A gas chromatography tandem mass spectrometry assay was used to determine estrogen components. RESULTS: The peak plasma concentrations of unconjugated and total estrone and equilin, unconjugated 17 beta-dihydroequilin and 17 beta-estradiol were higher and occurred earlier with the generic conjugated estrogens than with Conpremin. The 90% confidence limits for both variables lay outside the accepted bioequivalence limits of 80-125%. Additionally, no measurable plasma concentration of unconjugated delta 8,9-dehydroestrone or 17 beta-delta 8,9 dehydroestradiol was seen after administration of the generic conjugated estrogens. CONCLUSIONS: These pharmacokinetic results indicate that the generic tablets do not have the modified-release characteristics of Conpremin tablets. In addition, the absence of delta 8,9-dehydroestrone and 17 beta-delta 8,9 dehydroestradiol in the plasma indicates that the generic form is not compositionally equivalent to Conpremin. PMID- 11907936 TI - Phytoestrogens: dietary intake and excretion in postmenopausal Chinese women. AB - OBJECTIVES: The aims of this study were to estimate the dietary intake of phytoestrogens and to measure urinary phytoestrogen excretion in postmenopausal Chinese women. METHODS: Postmenopausal Chinese women were recruited from the hormone replacement clinic of the Prince of Wales Hospital. Dietary intake of phytoestrogen-rich foods was estimated using a food frequency questionnaire. Urinary output of the isoflavonoids daidzein and genistein and the metabolite of daidzein, equol, was measured using high-performance liquid chromatography. RESULTS: The mean daily excretion of daidzein, genistein and equol was 3.24 (+/- 3.63), 1.47 (+/- 1.75) and 0.64 (+/- 1.53) mumol, respectively. The total mean daily isoflavonoid excretion was 5.36 (+/- 5.27) mumol. CONCLUSIONS: Urinary excretion of isoflavonoid phytoestrogens in this Chinese population was lower than that reported in Japanese subjects. This may be due to the higher consumption of legumes, especially soy products, in the Japanese compared to the Chinese diet. The intake of green vegetables was higher in the Chinese subjects, and this food group may be the main contributor to their total phytoestrogen intake. PMID- 11907937 TI - Estrogens and the cardiovascular system: role of estradiol metabolites in hormone replacement therapy. AB - Estrogen substitution in the postmenopause reduces cardiovascular disease by means of direct and indirect effects of estradiol on the cardiovascular system. Recently, there have been increased indications that estradiol metabolites can also have beneficial effects. In the present short review, the existing experimental data for effects of estradiol metabolites on the blood vessels have been compiled. Results of our own studies, together with those of other research groups, indicate that particularly the catechol estrogens are able to exert a positive influence on the cardiovascular system, and may perhaps play a physiological role there. Attention is drawn to clinical-pharmacological aspects for use of estradiol metabolites for the prevention and treatment of cardiovascular disease. PMID- 11907938 TI - Phytoestrogens and the menopause. AB - Phytoestrogens are defined as naturally occurring plant compounds that are structurally and functionally similar to 17 beta-estradiol or that produce estrogenic effects. The commonest sources are cereals, legumes and grasses. Isoflavones are the most highly investigated subgroup of phytoestrogens. They are attenuated estrogens and behave both in vivo and in vitro as agonists and antagonists. The highest concentrations are found in soy beans and legumes. The relative potencies of isoflavones as compared to estradiol are small but they can exhibit bioactivity when tested in high concentrations. A high dietary intake of phytoestrogens was first noted to be associated with a decreased incidence of certain diseases. This epidemiological information was obtained primarily from studying Asian populations. Soy consumption is highest in Japan, where urinary levels of phytoestrogen metabolites are extremely high, and where there are lower rates of so-called 'Western' diseases, namely breast, endometrial, colon and prostatic cancers as well as atherosclerotic disease. These observations have prompted extensive research, which has demonstrated the varying degrees of estrogenicity of these phytoestrogen compounds. This article provides an epidemiological background to phytoestrogens, a brief description of their composition and biochemistry, and an overview of the literature to date on phytoestrogens with an emphasis on relief of menopausal symptoms. PMID- 11907939 TI - Management problems. BRCA mutation, prophylactic oophorectomy and HRT. PMID- 11907940 TI - HERS--a missed opportunity. PMID- 11907941 TI - Effect of hormone replacement therapy on self-reported cognitive symptoms: results from the Postmenopausal Estrogen/Progestin Interventions (PEPI) trial. AB - OBJECTIVE: To describe self-reported cognitive changes in 875 postmenopausal women aged 45-64 years and assigned randomly to hormone therapy (either estrogen alone or estrogen in combination with a progestin). METHODS: A 3-year placebo controlled trial which included self-report data on problems with forgetfulness, concentration and distraction in groups of women assigned randomly to placebo, estrogen alone or estrogen plus one of three progestin regimens. RESULTS: Women assigned to any active treatment were consistently less likely to report cognitive symptoms than women assigned to placebo, but these results were not statistically significant. When compared with women assigned to estrogen plus any progestin, women assigned to estrogen alone had a significant increased risk for reporting problems with forgetfulness at follow-up (odds ratio, OR = 1.47). In the absence of problems with distraction at baseline, women assigned to estrogen alone were significantly less likely to report problems with distraction at follow-up, compared to women assigned to estrogen plus any progestin (OR = 0.36). Assignment to a continuous progestin regimen in combination with estrogen was marginally associated with an increased risk for reporting concentration problems at follow-up, compared to cyclic progestin regimens (OR = 1.63). CONCLUSIONS: These results provide some evidence to support the hypothesis that estrogen improves memory, but findings are not statistically significant in this healthy middle-aged cohort. These data suggest that estrogen in combination with any progestin confers a decreased risk for reporting forgetfulness at follow-up, compared to estrogen alone. However, there is evidence that distraction problems reported at follow-up were side-effects of progestin. There is also some evidence that women administered progestins cyclically were at lower risk for reporting cognitive symptoms at follow-up, particularly concentration problems. PMID- 11907943 TI - Apolipoprotein E phenotype associations with plasma lipoproteins and bone mass in postmenopausal women. AB - OBJECTIVE: The aim of the study was to investigate the effects of apolipoprotein E (apo E) phenotype on plasma lipids and bone mass in postmenopausal Japanese women. METHODS: In 320 subjects aged 40-65 years (mean +/- SE, 54.0 +/- 0.6), apo E phenotype was determined by isometric electrophoresis. Phenotypic frequencies were 0.3% for E2/2 (n = 1), 8.1% for E3/2 (n = 26), 70.3% for E3/3 (n = 225), 0.6% for E4/2 (n = 2), 19.4% for E4/3 (n = 62) and 1.3% for E4/4 (n = 4). Apo E2/2 and apo E3/2 were classified as E2+ (n = 27); apo E3 homozygotes were placed in another group (E3/3; n = 225), as were apo E4/3 together with apo E4/4 subjects (group E4+; n = 66). Bone density and bone quality were assessed with a newly developed ultrasonic bone densitometer, and plasma lipids were also measured. RESULTS: Age, body mass index and years since menopause did not differ significantly between the three groups. The E4+ group had the highest levels of total cholesterol, low-density lipoprotein cholesterol and apolipoprotein B, significantly higher than in the E2+ group. Plasma lipoprotein(a) concentrations were significantly higher in the E4+ group than in the other two groups. Ultrasonic parameters of bone density and quality tended to be lower in the E4+ group than in the E2+ group. CONCLUSIONS: The apo E4 allele was associated with high plasma cholesterol levels and an unfavorable change of bone structure in postmenopausal women. PMID- 11907942 TI - Uterine surveillance of asymptomatic postmenopausal women taking tamoxifen. AB - The aim of this study was to assess the ability of transvaginal sonography (TVS) and office hysteroscopy with sharp curettage to characterize the morphological changes in the uteri of asymptomatic postmenopausal women taking long-term tamoxifen for breast cancer. The overall acceptability of a single-visit screening clinic for these women was also evaluated. Fifty-eight women were recruited from patients undergoing regular follow-up at the Leicester Royal Infirmary for breast cancer. A single-visit clinic was acceptable to 94.8% of these women. Transvaginal sonography detected endometrial thickness of greater than 5 mm in 84.5% of cases, but there was no relationship between total tamoxifen exposure and endometrial thickness. Transvaginal sonography also detected uterine lesions such as fibroids and endometrial cysts in 34.5% of cases. Hysteroscopy detected the latter uterine lesions in 53.4% of cases, with three cases (5.2%) of endometrial polyps also being identified in these women. Sharp curettage sampling of the endometrium produced specimens sufficient for diagnosis in 84.5% of cases; 70.7% of specimens were reported as showing types of 'quiescent' endometrium with 13.8% of specimens showing 'active' endometrium. In the latter group, there was a case of complex hyperplasia detected and also a case with granulomatous endometritis. For each histopathological diagnosis identified, there was a wide range of endometrial thickness recorded by TVS. A single-visit screening clinic involving TVS and hysteroscopy with sharp curettage was acceptable to asymptomatic women taking tamoxifen. However, hysteroscopy was more effective than TVS in detecting endometrial lesions such as polyps, fibroids and cystic areas. Although TVS detected endometrial thickness greater than 5 mm in the majority of cases, there were no malignancies detected and, for each histopathological classification, there was a wide range of endometrial thickness associated. Thus, the isolated use of TVS is insufficient for screening the endometria of these women. PMID- 11907944 TI - Once and twice a week transdermal estradiol delivery systems: clinical efficacy and plasma estrogen levels. AB - We compared the clinical efficacy and circulating estrogen levels from two transdermal delivery systems, 'drug-in-adhesive' type, in 20 healthy postmenopausal women. Both patches, developed by Beta Pharmaceutical Laboratories in Argentina, deliver estradiol at a rate of 50 micrograms/day; the replacement frequency of system A (TrialSat) was twice a week and for system B (TrialSat LA) once a week. The women were treated for 180 days, in a continuous regimen, with additional oral medroxyprogesterone acetate 5 mg/day for 14 days of each cycle. Blood samples were taken at the end of the wearing period: the 3rd day for Group A and the 7th day for Group B, to determine levels of estradiol, estrone, non-sex hormone binding globulin (SHBG)-bound estradiol and SHBG. Both treatments had similar clinical efficacy and were well tolerated. Plasma estradiol levels were higher in Group A throughout the study, probably owing to the different sampling times. SHBG and non-SHBG-bound estradiol were unchanged in both groups. As there was a similar performance of both delivery systems, the 7-day patch may be preferable, and produce greater compliance. PMID- 11907946 TI - Do combinations of 1 mg estradiol and low doses of NETA effectively control menopausal symptoms? AB - OBJECTIVES: This study compared two continuous-combined hormone replacement therapy (HRT) formulations, 1 mg estradiol (E2)/0.25 mg norethisterone acetate (NETA) and 1 mg E2/0.5 mg NETA, with placebo, with regard to the efficacy for vasomotor symptom relief in menopausal women. METHODS: A total of 119 women aged 45-61 years with moderate and severe hot flushes and with amenorrhea for at least 3 months were randomly assigned to 12 weeks' treatment with 1 mg E2/0.25 mg NETA, 1 mg E2/0.5 mg NETA or placebo. The number and severity of hot flushes, as well as any vaginal bleeding, were recorded on a daily basis. The Kupperman Menopausal Index, Greene Climacteric Scale and visual analog scales for various symptoms were assessed before and after treatment. Subpopulation analysis according to menopausal status was performed. RESULTS: Both combinations significantly reduced the number and severity of hot flushes, compared to placebo. A reduction of approximately 85% in vasomotor symptomatology occurred in the two combination groups by week 4 of treatment, and this was further diminished throughout the study to approximately 97% reduction by week 12. At the end of the study, 85% of the women receiving 1 mg E2/0.5 mg NETA and 71% of the women receiving 1 mg E2/0.25 mg NETA were considered to the clinically adequate responders to treatment. Both combinations were associated with significant improvements, compared to placebo, in visual analog scales for overall general condition, Kupperman Menopausal Index, and Greene Climacteric vasomotor and psychological subscales. While both combinations resulted in similar bleeding profiles in postmenopausal women, the combination of 1 mg E2/0.5 mg NETA resulted in the lowest incidence of bleeding in late perimenopausal women. CONCLUSIONS: The combinations of 1 mg E2/0.25 mg NETA and 1 mg E2/0.5 mg NETA rapidly relieve vasomotor symptoms and are efficacious in the majority of menopausal women, including those with severe hot flushes. PMID- 11907945 TI - A 3-year study of prevention of postmenopausal bone loss: conjugated equine estrogens plus medroxyprogesterone acetate versus tibolone. AB - The aim of this study was to investigate the effects of tibolone in the prevention of postmenopausal bone loss over 3 years, and to compare these with the effects of sequential hormone replacement therapy. Forty early postmenopausal women were randomized to a 21-day regimen of conjugated equine estrogens (CEE, Premarin) plus sequential medroxyprogesterone acetate (MPA, Prodafem), or tibolone (Livial) daily. In total, 36 women completed 12 months and were considered for the intent-to-treat analysis, 34 completed 24 months and 23 completed 36 months. Main drop-out reasons were: lost to follow-up (n = 9) and minor side-effects (n = 4). Bone mineral density was measured at baseline and after 6, 12, 24 and 36 months, using dual-energy X-ray absorptiometry at the lumbar spine and the upper femur (neck, trochanter, total hip). In both groups, bone loss was prevented. Treatment with tibolone demonstrated significant increases in bone density at the spine (+4.6%; p < 0.01), at the total hip (+3.2%; p < 0.01) and at the trochanter (+4.5%; p < 0.01), whereas the CEE/MPA group showed a non-significant increase of bone mineral density at the lumbar spine (+2.6%) but no increases at the hip. Between-group differences in bone mineral density changes were significant (p < 0.05) for the total hip and the trochanter at 36 months. This increase of bone mineral density was not accompanied by changes in insulin-like growth factor-I (IGF-I) or insulin-like growth factor binding protein-3 (IGFBP-3) in either group. Osteocalcin, alkaline phosphatase and urinary ratios of hydroxyproline/creatinine and calcium/creatinine significantly decreased in both groups. In conclusion, sequential CEE/MPA prevented cortical and trabecular bone loss, with a transient increase of bone mineral density only during the first 6 months. Tibolone not only prevented cortical and trabecular bone loss, but further increased bone mineral density at the lumbar spine and at the hip throughout the 3 years of treatment, suggesting a sustained positive effect on bone mass. PMID- 11907947 TI - How significant are environmental estrogens to women? AB - Women are exposed to xenobiotic estrogens at least to the same extent as men. These estrogenic chemicals are either from plant material in the diet (phytoestrogens) or from industrial sources. Mainly industrially derived environmental estrogens may accumulate within the food chain and persist in human adipose tissue. In contrast, phytoestrogens do not bioaccumulate and are rapidly excreted in urine. The phytoestrogens probably represent the source of most extensive exposure for humans. Epidemiological evidence suggests that diets rich in phytoestrogens are associated with reduced incidences of cardiovascular disease, breast cancer, prostate cancer and osteoporosis. The numerous bioactivities (other than just estrogenicity) of phytoestrogens and related dietary compounds make it difficult to single out the mechanisms mediating such protective effects. The possibility that the newly discovered estrogen receptor beta may be an important modulator of phytoestrogen action is opening up new lines of research. While the evidence suggests that phytoestrogens may be of positive relevance to postmenopausal women, indications that exposure of women to industrially derived xenobiotic estrogens provides risks to health remain unproven. Further work is necessary to clarify the relative importance of 'xenobiotic' estrogens to human health, but it must be emphasized that the estrogenic potency of all the xenobiotic estrogens is very low compared with that of endogenous estrogens. PMID- 11907948 TI - Phytoestrogen study design. PMID- 11907949 TI - [Pheochromocytoma in children]. AB - Pheochromocytoma is rare in children. A wide varieties of lesions are observed and diagnosis, often made late, is based on urinary catecholamine assay. Magnetic resonance imaging provides the best morphological information. The disease is usually benign and prognosis is good. Familial forms of pheochromocytoma and pheochromocytoma associated with ischemia-induced bone lesions must be emphasized. PMID- 11907950 TI - [Autoimmune hepatitis. Physiopathologic, clinical, histological, and therapeutic features]. AB - Autoimmune hepatitis is characterized by an inflammation of the portal tract with lymphocytes and plasma cells, an hypergammaglobulinemia and a variety of circulating autoantibodies. The presence of smooth muscle antibodies and/or antinuclear antibodies define type 1. Type 2 is characterized by the presence of liver-kidney--microsomal antibodies. Environmental, genetic and infectious factors may explain the autoreactivity of T cells. Different non specific clinical features may be present. Sometimes the presentation may be an acute hepatitis; in the remainder, the disease may not be recognized until liver damage is advanced. Hypergammaglobulinemia and presence of circulating autoantibodies are the key for diagnosis. The association of prednisolone in combination with azathioprine remains the established treatment. If relapse or non response occur, other immunosuppressive therapy such as cyclosporin may be useful. Liver transplantation is reserved for (sub)fulminant forms and end stage liver disease. PMID- 11907952 TI - [Vitamin E deficiency. Etiopathogenesis, clinical, histopathologic, and electrical features, and main etiologies]. AB - Vitamin E deficiency is frequently observed in human pathology. In most cases, deficiency is moderate and asymptomatic. Severe deficiency is rare and presents as a progressive neurological syndrome including cerebellar ataxia and posterior cord injury. Neuropathological and electrophysiological features confirm spinocerebellar degeneration. The pathophysiology of vitamin E deficiency nervous dysfunction is still unknown. Oxidative alterations due to the lack of the main lipid-soluble antioxidant could be involved. A few causes of vitamin E deficiency are recognized (malnutrition, impaired lipid absorption, specific anomaly of hepatic or intestinal secretion of vitamin E, excessive endogenous consumption), but diseases associated with vitamin E deficiency are numerous and divers. Symptomatic severe deficiency is constantly observed in ataxia with vitamin E deficiency and abetalipoproteinemia. Intestinal, hepatobiliary, and pancreatic diseases are often associated with more or less marked deficiency. Other pathological circumstances such as malnutrition, alcoholism, hemolytic anemia, renal failure and hemodialysis could be associated with moderated and asymptomatic vitamin E deficiency. Oral or regular administration of high-dose vitamin E is required for patients with symptomatic severe deficiency. An adapted supplementation is recommended for patients with a marginal vitamin E status. PMID- 11907951 TI - [Vitamin E: structure, metabolism, and functions]. AB - Vitamin E is a generic term denoting eight different isomers among which alpha tocopherol is the most important and most active. Vitamin E metabolism is closely linked to lipids during intestinal absorption, plasma secretion and transport, and tissue uptake. It is a key compound involved in many physiological processes, such as neurological and immune functions. The most common role of vitamin E is its antioxidant effect, protecting molecules and tissues against the deleterious effect of free radicals. Vitamin E also contributes to the stabilization of biological membranes. In addition, it intervenes in the regulation of several enzymes and probably has impact on gene expression. Advancing knowledge about vitamin E has been achieved with high performance liquid chromatography, making assay accessible to many laboratories, and the use of deuterated derivatives to better apprehend its metabolism. Certain issues remain unresolved concerning the molecular basis of vitamin E's mechanism of action and the exact nature of metabolic dysfunction leading to the clinical manifestations of severe vitamin E deficiency. PMID- 11907953 TI - [Vitamin E deficiency: risk factor in human disease?]. AB - Oxidative stress is suspected to intervene in certain chronic diseases. Much research has been devoted to the relationship between vitamin E, a major lipid soluble antioxidant, and certain widespread diseases. Experimental and epidemiological proof supports a protective effect of vitamin E in a number of pathological processes such as coronary heart disease, cancer, cataract, diabetes mellitus, and Alzheimer disease. Randomized clinical trials have not confirmed a beneficial effect of vitamin E supplementation on the progression of these diseases. Certain methodological biases could however have affected these studies, explaining conflicting results. These biases include inaccuracy of vitamin E intake estimates and changes in eating habits during the course of the survey. An insufficient supplementation period using an insufficient dose and inclusion of aged and high-risk patients are the main limitations of the reported clinical trials. Large scale randomized clinical trials including healthy and low risk subjects, along with prolonged administration of high-dose natural vitamin E, associated with synergetic compounds, and testing on morphological or biological features, will allow a better understanding of the relationship between vitamin E and chronic diseases. PMID- 11907954 TI - [Acute community acquired pneumopathy caused by Nocardia asteroides in a 93-year old female patient]. AB - BACKGROUND: Lower respiratory tract infection (LRTI) by Nocardia species is most often considered as an opportunistic infection occurring mainly in middle-aged adults. CASE REPORT: A 93-year old woman without any significant comorbidity other than a mild asthma, presenting with a non-resolving pneumonia, was successfully treated for a community-acquired LRTI by Nocardia asteroides. DISCUSSION: LRTI by Nocardia asteroides is a rare occurrence. Although more frequent in immuno-compromised patients (50 to 85% of cases reported), Nocardia asteroides infection also occurs in immuno-competent individuals. Very few reports describe LRTI infection by Nocardia sp. in the very old, and almost exclusively in patients with major co-morbidities or variable degrees of immuno suppression. PMID- 11907955 TI - [Peri-aortic fibrosis: unusual case of jaundice]. AB - Periaortic fibrosis is defined as the development of aortic perianeuvrysmal fibrosis either resulting from a focal inflammatory reaction or a self perpetuating process. Compression of neighboring organs is a possible complication. Bile duct obstruction is exceptional. PMID- 11907956 TI - [Intestinal lymphoma associated with Behcet disease]. AB - The association of Behcet's disease and non-Hodgkin's lymphoma is very rare. The first case of primary intestinal lymphoma with intestinal Behcet's disease is reported. A 37 year-old woman had been treated for Behcet's disease with colchicine for 2 years. In July 1997, she developed uveitis and was treated by monthly intravenous pulses of cyclophosphamide for 6 months, but uveitis persisted; so cyclophosphamide was replaced by cyclosporin 300 mg/day. One month later, she suffered from diarrhea. Colonoscopy showed ileocoecal ulcerations. Histological examination of surgical biopsy revealed B large cell type lympocytic malignant lymphoma and vasculitis lesions compatible with intestinal Behcet's disease. Cyclosporin was stopped and treatment with prednisone was instituted. The relationship between non-Hodgkin's lymphoma and immunosuppressive drugs in Behcet's disease is discussed. PMID- 11907957 TI - [News of the Association of French speaking dermatologists. Memories from Cayenne]. PMID- 11907958 TI - ["Idiopathic" white atrophy]. AB - INTRODUCTION: Livedoid vasculitis is a clinico-pathological entity which may be idiopathic or secondary to various disorders. The aim of this study was to search for a thrombogenic biological abnormality in patients with apparently idiopathic livedoid vasculitis. PATIENTS AND METHODS: All the patients with histologically confirmed and apparently idiopathic livedoid vasculitis were evaluated. Blood study included search for anticardiolipin, anti-anionic phospholipids and anti beta 2 glycoprotein 1 antibodies, platelet aggregation and fibrinolytic system tests, cryofibrinogen and homocysteine serum level and factor V Q506 mutation and prothrombin 20210 G/A variant investigation. Clinical data and effects of treatments were reviewed retrospectively. RESULTS: Eleven of the 21 patients with livedoid vasculitis had an apparently idiopathic form. Seven of them (64 p. 100) had a thrombophilic state: antiphospholipid antibodies (n = 3), increased platelet aggregation (n = 1), cryofibrinogen (n = 1), decreased antithrombin III activity (n = 1) and factor V mutation (n = 1). The necrotic lesions were always localized on the lower limbs with a sensitive neuropathy in 2 cases. Complete remission was sometimes obtained with antiaggregant or anticoagulant therapy, but was unrelated to the thrombophilic abnormalities. CONCLUSION: Various thrombophilic abnormalities are frequently observed in livedoid vasculitis which seems to be the clinical expression of a thrombotic process of the microcirculation of the skin and sometimes of the peripheral nerves. The idiopathic feature of numerous cases gives evidence for the lack of our knowledge on thrombosis of the microcirculation. PMID- 11907959 TI - [Primary cutaneous cryptococcosis in HIV-seronegative subjects]. AB - BACKGROUND: Primary cutaneous cryptococcosis is an uncommon clinical entity characterized by direct skin inoculation without systemic involvement. We report four cases of this affection in HIV-negative patients seen between 1990 and 1999 in Nantes Dermatological Clinic. CASE REPORTS: Patients mean age was seventy. Three patients had recent exposure to soil or birds, and two remembered a trauma before the lesion appeared. In three cases the lesion was on the hand. In two cases the lesion was an ulcerated nodule, in another an abscess and in the last a cellulitis. Two subjects were treated by fludarabin and systemic corticosteroids for respectively chronic lymphocytic leukemia and Waldenstrom's macroglobulinemia. The third had a CD4 lymphopenia. Cultural examination confirmed the diagnosis. Treatment, with fluconazole in 3 cases and ketoconazole and itraconazole in 1 case, resulted in healing of the skin lesion in a few months. DISCUSSION: Recognition of primary cutaneous cryptococcosis as a clinical entity has long been debated. An altered immunological status is an important factor for developing this disease. There is often a clear history of trauma or exposure to soil or birds preceding the development of the lesion. Clinically it often looks like a papule or an ulcerated nodule. The lesion is confined to the skin without systemic involvement. The prognosis is excellent thanks to the use of oral antifungal imidazoles. PMID- 11907960 TI - [Drug hypersensitive syndrome caused by fluindione]. AB - BACKGROUND: Fluindione (Previscan) is an oral anticoagulant prescribed in relay to heparin therapy for deep venous thrombosis, pulmonary embolism... The main complications with oral anticoagulants are bleeding. However, severe immuno allergic complications, especially acute hepatitis, acute renal failure and acute bone marrow failure, have been described with phenindione therapy. OBSERVATIONS: The authors report herein the first five cases of drug-induced hypersensitivity syndrome due to fluindione. Clinical signs included erythroderma and severe systemic manifestations which occurred within 3 to 8 weeks after introducing the molecule. Sex ratio was four males for one female; their ages ranged from 53 to 84 years. Clinical signs included erythroderma (with photosensitivity in two patients), lymphadenopathy and fever evoking severe sepsis. In our observations, marked eosinophilia (5 cases), lymphocytosis, atypical lymphocytes (4 cases), hepatic cytolysis (4 cases), associated in 2 cases with hepatic cholestasis, and pulmonary signs were noted. Cutaneous eruption healed in about 3 to 6 weeks after withdrawal of the drug. In two cases, systemic steroids were required for the severity of systemic manifestations. Long after the acute episode and when steroids were stopped, patch testing with fluindione was still positive. DISCUSSION: To date, acute and/or severe skin diseases due to fluindione, associated or not with multisystemic involvement, have never been reported. This molecule is the most commonly used for the treatment of thromboembolic diseases. Patch testing is easy to perform and can help physicians find the responsible molecule. Moreover, skin manifestations are present only in 87 p. 100 of drug induced hypersensitivity syndromes. Acute hepatitis and acute renal failure might be drug-induced hypersensitivity syndromes without cutaneous manifestations. PMID- 11907961 TI - [Photoaggravated contact allergy and contact photoallergy caused by ketoprofen: 19 cases]. AB - INTRODUCTION: Between September 1994 and September 1999, we observed 19 cases of photoaggraved contact allergy or contact photoallergy to ketoprofen (non steroidal anti-inflammatory derived from arylpropionic acid). We present a clinical and photobiological retrospective study of these 19 cases with investigation of cross-reactivity between benzophenone-containing molecules. PATIENTS AND METHODS: On clinical level, we investigated the type of eruption, the delay of appearance, the initial area of eruption and areas of diffusion. Phototesting included patchtests and photopatchtests performed with the gel containing ketoprofen (17 patients), ketoprofen 2 p. 100 petrolatum (14 patients), fenofibrate 10 p. 100 petrolatum and 10 p. 100 water (15 patients), 3 benzophenones (19 patients): oxybenzone 10 p. 100 petrolatum, mexenone 2 p. 100 petrolatum, sulisobenzone 10 p. 100 petrolatum and the other arylpropionic derivatives (4 patients). Three identical series were applied: one was irradiated with 3/4 polychromatic minimal erythematosus dose, a second was irradiated with UVA 13 J/cm2 until January 1997, then 5 J/cm2, the third series was not irradiated (control series). RESULTS: Patients were 9 men and 10 women with an average age of 41.2 years. The type of eruption was an eczema. The delay of appearance of the eruption was one day to 3 months. For 10 patients, the delay was between 4 and 18 days. The eruption was localized to the application area in 1 case, to the application area then to the same contralateral area in 3 cases, to the application area then to all photoexposed areas in 13 cases, to the application area then to the photoexposed areas and then to non-sun-exposed areas in 2 cases. Evolution showed prolonged photosensitivity in 3 cases after withdrawal of the contact and the contact photoallergy to ketoprofen was severe. Gel-containing ketoprofen photopatchtests showed 9 photoaggravated contact allergy, 6 contact photoallergy and 2 contact allergy. Ketoprofen photopatchtests showed 12 contact photoallergy and 2 photoaggraved contact allergy. Tiaprofenic acid photopatchtests were positive in all performed cases (4/4), but photopatchtests with the other arylpropionic derivatives, without benzophenone structure, were negative. Fenofibrate photopatchtests were always positive (15/15). Benzophenones photopatchtests only showed 4 cases of contact photoallergy to oxybenzone (4/19). In 68 p. 100 of cases, patients presented a contact allergy or photoallergy to fragrances. CONCLUSIONS: This study shows the actual frequency of contact allergy and contact photoallergy to ketoprofen with a higher frequency of contact photoallergy. Thus, photopatchtesting is essential. In cases of contact photoallergy to ketoprofen, ketoprofen, tiaprofenic acid but not the other arylpropionic derivatives, fenofibrate and benzophenones have to be withdrawn. PMID- 11907962 TI - [Absence of interaction between thiomersal and aluminum occlusion chambers]. AB - BACKGROUND: Positive patch tests to thiomersal are frequent, but of little relevance. Interaction between aluminium occlusive chambers and the mercury of thiomersal may elicit skin irritation and cause false positive results. METHODS: Between January 1998 and May 1999, the patients were simultaneously patch tested with aluminium and plastic chambers. Positive skin tests were compared. RESULTS: Seventeen of 213 subjects showed positive tests: 15 with the 2 materials, 1 only for the aluminium chamber, 1 for the plastic chamber. DISCUSSION: The results did not differ for the two materials. We concluded that positive reactions observed were not caused by interaction between aluminium and thiomersal. PMID- 11907963 TI - [Skin manifestations of immune restoration syndrome in treated tuberculosis]. AB - INTRODUCTION: Immune restoration syndrome was first described in 1998 and involved mycobacterium avium complex. We report the case of a patient with acquired immunodeficiency syndrome who had disseminated cutaneous lesions due to Mycobacterium tuberculosis, following initiation of highly active antiretroviral therapy. CASE REPORT: A 42 year-old HIV-infected man, was admitted for fever, cough, nocturnal sweat and impaired of general condition. He had a viral load of 127,200 copies/ml and 199/ml CD4 T-cells. He was treated with triple tuberculosis combination therapy according to tuberculous contagium, positivity of the tuberculin intradermoreaction (15 mm) and right upper lung nodule on thoracic scan. M. tuberculosis was not found. Fever improved at day 3. Highly active antiretroviral therapy with zidovudine, lamivudine, indinavir, was started at day 11 and 33 days after, fever and dermohypodermal nodules with necrotising evolution appeared. Skin biopsy specimen showed tuberculoid granuloma. The levels of viral load and CD4 T-cells were less than 200 copies/ml and 497/ml respectively. Fever and cutaneous lesions spontaneously resolved without changing therapy. DISCUSSION: Immune restoration syndrome appears after initiation of antiretroviral therapy, in patients with advanced HIV infection and without prophylactic treatment versus MAC. This case report probably involves mycobacterium tuberculosis. Bacterial lysis and immune restoration take part in cutaneous pathogenesis. Subclinical mycobacterial infection should be monitored during initiation of antiretroviral therapy in patients with advanced HIV infection. PMID- 11907964 TI - [MELAS syndrome (mitochondrial encephalopathy with lactic acidosis and stroke like episodes]. AB - BACKGROUND: The MELAS syndrome (Mitochondrial Encephalopathy, Lactic Acidosis and Stroke-like episodes) belongs to the category of mitochondrial disorders. The most common molecular etiology of the syndrome is a mutation A to G transition at base pair 3243 in the mitochondrial genome. The phenotype is varied and depends on the proportion of DNA muted and which organ on aerobic metabolism suffers most. CASE-REPORT: An 17 year-old woman had successively neurosensory hearing loss, renal disease, cardiomyopathy, diabetes mellitus, lactic acidosis and stroke-like episodes that evoked a MELAS syndrome. DISCUSSION: The skin manifestations of patients with MELAS syndrome are scaly, pruritic, diffuse erythema, reticular pigmentation, moderate hypertrichosis, seborrheic eczema, atopy and vitiligo. Our patient presented severe hirsutism and reticular pigmentation of the limbs. No abnormal histologic and electron microscopic findings were noted in the skin or the follicles involved. PMID- 11907965 TI - [Pseudoainhum in lamellar ichthyosis]. AB - INTRODUCTION: Pseudoainhum is an affection characterized by the appearance of a constricting band around a digit or limb which may lead to spontaneous amputation. There are various etiologies which can be either congenital or resulting from a concomitant disease. We report herein an original case which occurred during lamellar ichthyosis. CASE-REPORT: We present the case of a thirty year-old woman affected by a severe form of lamellar ichthyosis who, in a few months, developed a pseudoainhum of the third left finger. Biological results were normal. X-rays of this finger showed distal resorption of the bone. The painful evolution and the absence of any conservative treatment compelled us to propose amputation of the third phalanx. DISCUSSION: Other cases of pseudoainhum have been observed in many disease responsible for vascular or neurological abnormalities. They can also result in physical traumatisms or be associated with many dermatoses, such as hereditary palmar and plantar keratodermas. As far as we know, this lesion has never been described during lamellar ichthyosis but could result in the keratinization disorders observed in this congenital ichthyosis. PMID- 11907966 TI - [Severe pustular and polymorphous vasculitis caused by losartan]. AB - INTRODUCTION: We report a case of severe pustular vasculitis most probably due to losartan (Cozaar) which is an antihypertensive drug inhibiting the receptors for angiotensin II. CASE REPORT: A 64-year-old woman, suffering from hypertension, developed a widespread and rapidly evolving bullous and pustular eruption without any other clinical sign. Leucocytoclastic vasculitis and junctional blisters were present with deposits of both IgA and IgG at these sites. Withdrawal of losartan, which was the only drug taken by the patient, and tapered corticotherapy cleared the lesions. The rash left a prominent mottled hyperpigmentation. It did not recur during the following months despite the presence of antineutrophil cytoplasmic antibodies. DISCUSSION: So far only a few cutaneous side effects have been reported following losartan intake. This drug was the likely culprit in the present case. Vasculitis was extremely severe and polymorphous. PMID- 11907967 TI - [Allgrove's syndrome]. AB - INTRODUCTION: Triple A syndrome is an autosomal recessive disease causing achalasia, alacrima and adrenal involvement with isolated glucocorticoid deficiency. Less than 70 cases have been reported worldwide. We report a case of familial adrenal insufficiency with hyperpigmentation diagnosed in a 14 month-old child. CASE REPORT: A 2 year-old boy, a product of consanguineous parents, was referred to our institution for evaluation of melanoderma. Since birth the boy had suffered from vomiting and diarrhea. Aged 6 months, mucosal erosive lesions had appeared associated with fever and further complicated at the age of one year by alacrima. At the age of 14 months, hyperpigmentation of all the skin had occurred. The family history is significant: two siblings (a girl and a boy) died of hypoglycemia with melanoderma and alacrima at the age of 5 and 3 respectively. Physical examination showed generalized hyperpigmentation particularly marked on the lips and genitalia. The genitalia of our patient were normal. Cortisol was 23.7 microns/l (normal: 193-772); ACTH was 11,722 pg/ml (normal: 10-60) and computed tomography of the abdomen confirmed adrenal gland hypoplasia. Treatment was initiated with hydrocortisone at the dose of 10 mg/day. At the age of 3, the boy developed plantar hyperkeratosis. When solid food was introduced, vomiting and regurgitation increased. An oesophagogram with fibroscopy revealed achalasia of the cardia. This was successfully corrected by surgery. Schimer's test confirmed alacrima. DISCUSSION: Our case report, characteristic of triple A syndrome, is unusual in view of the presence of plantar keratoderma and absence of any neurological abnormalities. PMID- 11907968 TI - [Imported cutaneous leishmaniasis caused by Leishmania infantum confirmed with immunoassay]. AB - BACKGROUND: Localized cutaneous leishmaniasis acquired in France is rarely reported. Diagnosis usually relies on detection of leishmania on smears or by culture. CASE REPORT: A 9-year-old child living outside endemic French Mediterranean areas had long-lasting crusted papules on the face for several months. Although the lesions were suggestive of cutaneous leishmaniasis, smears and culture were negative for Leishmania. Skin biopsy showed epithelioid and giant cell granuloma, but Leishman bodies were absent. Western Blot analysis of the patient's serum revealed antibodies directed against Leishmania infantum antigens, thus confirming the diagnosis of cutaneous leishmaniasis. Intralesional injections of meglumine antimoniate yielded complete regression of lesions. DISCUSSION: Localized cutaneous leishmaniasis in France is caused by Leishmania infantum and may be diagnosed outside endemic Mediterranean areas, following transmission from a sandfly bite during summer holidays in Southern France. Serum analysis by Western Blot assay distinguishes between clinically active and asymptomatic Leishmania infections, the latter being common in endemic areas. Western Blot analysis is useful for the diagnosis of cutaneous leishmaniasis when parasites cannot be detected by direct techniques. PMID- 11907969 TI - [Bilateral orbital lipomatosis in systemic cortical therapy]. PMID- 11907970 TI - [Skin and bone lesions]. PMID- 11907971 TI - [Brown spots in a newborn]. PMID- 11907972 TI - [Imported cutaneous leishmaniasis in metropolitan France]. PMID- 11907973 TI - [Hereditary osteo-onychodysplasia (nail patella syndrome)]. PMID- 11907975 TI - [Frostbite]. PMID- 11907974 TI - [Porphyria cutanea tarda]. PMID- 11907976 TI - [Areolo-mammary hyperkeratosis]. PMID- 11907977 TI - [Spontaneous repigmentation of white hair]. PMID- 11907980 TI - [How can anesthetic accidents studied better?]. PMID- 11907981 TI - [Photodynamic therapy with 5-aminolevulinic acid or placebo for recalcitrant foot and hand warts: randomised double-blind trial]. PMID- 11907982 TI - [Annales de Dermatologie in English]. PMID- 11907983 TI - [Risk factors of melanoma in mediterranean populations]. PMID- 11907984 TI - [Immune restoration syndromes in human immunodeficiency virus infection]. PMID- 11907985 TI - [Urticaria]. PMID- 11907986 TI - [Mediators causing urticaria]. PMID- 11907987 TI - [Eosinophils and urticaria]. PMID- 11907988 TI - [Food allergy and urticaria]. PMID- 11907989 TI - [Airborne allergen induced urticaria]. PMID- 11907990 TI - [Airborne and food allergen cross reactions]. PMID- 11907991 TI - [Chronic urticaria]. PMID- 11907992 TI - [Urticaria and systemic diseases]. PMID- 11907993 TI - [Contact urticaria]. PMID- 11907995 TI - Drug-related morbidity and mortality: an economic and clinical perspective. PMID- 11907994 TI - [Drug-induced urticaria]. PMID- 11907996 TI - Healthcare's uncertain future. PMID- 11907997 TI - Ice bound. PMID- 11907998 TI - DTC (direct-to-consumer): a powerful tool for information, education. PMID- 11907999 TI - Tuesdays with Morrie. PMID- 11908000 TI - Demographic shifts--the age wave. PMID- 11908002 TI - WA nurse blows whistle on aged care. PMID- 11908001 TI - Health insurance membership drops again. PMID- 11908003 TI - Practice nurses boosted by budget. PMID- 11908004 TI - Radiation oncology inquiry announced. PMID- 11908005 TI - District nurses get IT support. PMID- 11908006 TI - Fifty families have their say. PMID- 11908007 TI - Maximising the benefits of information. Interview by Steven Harulow. PMID- 11908009 TI - What about our rights? PMID- 11908008 TI - Quality must be enshrined. PMID- 11908010 TI - Better use of nurses reduces waiting times. PMID- 11908011 TI - I.V. fluids and microwaves oven safety. PMID- 11908013 TI - Report supports nurses in triage role. PMID- 11908012 TI - Nurses and the Glasgow coma scale. PMID- 11908014 TI - Violence: the role of training. PMID- 11908015 TI - Technology: but not at the expense of care. PMID- 11908016 TI - No rivalry or animosity: just mutual respect. PMID- 11908017 TI - Persevering with the physics and chemistry. PMID- 11908018 TI - Transport noise stresses sick babies. PMID- 11908020 TI - Rally signals aged care crisis as election issue. PMID- 11908019 TI - Nurses don't want to go home. PMID- 11908021 TI - Report shows Bishop wrong on aged care funding. PMID- 11908022 TI - Nurse numbers falling: workloads rising. PMID- 11908023 TI - Sydney University cuts nurse training. PMID- 11908025 TI - Vic nurses win world's first nurse-patient ratios. PMID- 11908024 TI - ANF supports scholarships. PMID- 11908026 TI - ANF makes submission on nursing to Senate inquiry. PMID- 11908027 TI - Communication between clients, carers and staff in a dementia care facility. PMID- 11908028 TI - Effect of health education on patients' beliefs about glaucoma and compliance. AB - A pretest-posttest control group experimental design (n = 100) was used to determine the effectiveness of an interactive patient education program compared with a didactic approach for persons with primary open angle glaucoma at a major specialist eye hospital in England. This study used a questionnaire with a knowledge test to explore patients' glaucoma knowledge, a series of vignettes to explore understanding of compliance and health motivation, and health locus of control scales to assess the effect of these variables. The improved posttest results (P = .000) suggest that patients benefit from education programs and that the ophthalmic nurse is an effective patient teacher. The interactive program has no statistically significant difference from the didactic presentation. Other types of interactive programs may prove to be more beneficial. PMID- 11908029 TI - Look on the bright side: a case study of uveitis. AB - This article explores the contemporary approach to the care of patients with uveitis, an inflammation of the uvea or middle layer of the eye, including the etiology, symptoms, diagnosis, types, and contemporary treatment of this serious ophthalmic condition. Uveitis represents the third-leading cause of blindness in developed countries. Vision loss with this disease occurs usually as a result of very slow damage to the macula, which is the consequence of low-grade chronic inflammation. A case study involving a young man with uveitis is used to demonstrate the importance of treating active inflammation aggressively rather than waiting for a particular level of degradation. The role of the professional nurse is highlighted because the support, care, and education of this patient appear to be major components to his well being. PMID- 11908030 TI - Compensating for sight loss with a guide dog. AB - Although medical advances occur daily, ophthalmic nurses face vision decrease or loss in their patients. Orientation and mobility training are offered by state agencies, helping the person find ways to adjust to living with sight loss. Whereas a white cane is the preferred mobility method of choice for some persons, more persons are discovering the advantages of a guide dog. This article will help ophthalmic nurses understand how a guide dog is produced and the uses of a guide dog. This knowledge is an option to increase mobility in newly blind patients. PMID- 11908031 TI - The colored lenses controversy. PMID- 11908032 TI - Dry eye: an introduction for ophthalmic health care personnel. PMID- 11908033 TI - Anti-amalgam commentary "it seems to me...". PMID- 11908034 TI - ADA briefs. Independent contractor or employee: be careful! PMID- 11908035 TI - ADA briefs. Appearance of patient records is important. PMID- 11908036 TI - Professional office sharing agreements. The big wave of the future. PMID- 11908038 TI - Children's dental health month: more important than ever. PMID- 11908039 TI - Children's dental health month in Hawaii. PMID- 11908040 TI - Humana goes online to promote patient choice in benefit plans. AB - For Humana, one way to promote quality care is to provide choice among health care consumers by using online resources. Last fall, the plan began rolling out Emphesys, an new online interactive health insurance plan, in individual markets across the country. PMID- 11908041 TI - Leapfrog Group patient safety initiatives draw kudos, criticism. AB - Just over a year ago, the Leapfrog Group proposed a three-pronged patient safety initiative. One year later, the group is finding that many hospitals and health systems across the country are looking closely at the recommendations and making changes. But, it also is finding that it has become a lightning rod within the health care community in debates on whether these proposals are the best way now to proceed in improving patient safety. PMID- 11908042 TI - Highlighting award-winning quality initiatives. AB - Recent award-winning hospitals and health systems share insights into their programs and initiatives that promote quality of care. While these programs address vast and different areas--using ACE inhibitors, expanding community services, providing mental health support, reducing nosocomial pneumonia, and enhancing patient safety--they all have one thing in common: Their work represent weeks, years, and even decades of extraordinary vision, planning, and just plain hard work to successfully provide quality care to patients and communities. PMID- 11908043 TI - Supportive communication in the face of breast cancer. PMID- 11908044 TI - Spiritual care. Lingering questions. PMID- 11908046 TI - Walking with fan through BMT. Bone Marrow Transplant. PMID- 11908047 TI - Rose Hawthorne Lathrop. Servant of the dying poor. PMID- 11908048 TI - Diane's legacy. PMID- 11908049 TI - Evelyn & Charles. An oasis of love in the ER. PMID- 11908050 TI - Does prayer really help? PMID- 11908051 TI - Heart surgery. Body & soul. PMID- 11908052 TI - Someone to love me. PMID- 11908053 TI - Trauma revisited. One nurse's response to terrorism. PMID- 11908054 TI - Evolution and diversity of prokaryotic small heat shock proteins. PMID- 11908055 TI - The developmental expression of small HSP. PMID- 11908056 TI - Expression and phosphorylation of mammalian small heat shock proteins. PMID- 11908057 TI - sHsp-phosphorylation: enzymes, signaling pathways and functional implications. PMID- 11908058 TI - Small stress proteins: modulation of intracellular redox state and protection against oxidative stress. PMID- 11908059 TI - Small stress proteins: novel negative modulators of apoptosis induced independently of reactive oxygen species. PMID- 11908060 TI - Discovery of two distinct small heat shock protein (HSP) families in the desert fish Poeciliopsis. PMID- 11908061 TI - Hsp27 as a prognostic and predictive factor in cancer. PMID- 11908062 TI - Cytoskeletal competence requires protein chaperones. PMID- 11908063 TI - Hsp27 in the nervous system: expression in pathophysiology and in the aging brain. PMID- 11908064 TI - Protection of neuronal and cardiac cells by HSP27. PMID- 11908065 TI - Chaperone function of sHsps. PMID- 11908066 TI - The small heat shock proteins of the nematode Caenorhabditis elegans: structure, regulation and biology. PMID- 11908068 TI - Roles of SCF and VHL ubiquitin ligases in regulation of cell growth. PMID- 11908067 TI - Drosophila small heat shock proteins: cell and organelle-specific chaperones? PMID- 11908069 TI - Proteasome inhibitors. PMID- 11908070 TI - Aging of proteins and the proteasome. PMID- 11908071 TI - Protein degradation in the aging organism. AB - It is now generally accepted that protein degradation declines with age but a mechanism of action for this decline has not yet been delineated. Although intracellular and extracellular proteins can enter multiple pathways of degradation, there primarily appears to be two final mediators of this degradation, the lysosome and the proteasome. Studies on the effects of age on lysosomal function suggest that, if anything, lysosomal enzyme activity increases with age (Ward 2000). The peptidase activities of the proteasome are altered with age, but not in a consistent manner. There is a significant age-related decline of the PGPH activity, but the rate-limiting peptidase activity, ChT-L activity, as well as T-L activity have both been reported either to increase, not change, or decrease (Table 1). In addition, proteasomal degradation of casein does not appear to be altered with age. As a result, it has not been possible to definitively implicate either of the two primary final mediators of protein degradation, the lysosome and the proteasome, as mechanisms of action for the decline in protein degradation observed in the aging organism. However, there are experimental observations suggesting that age may have strong effects on both macroautophagic and the chaperone-mediated autophagic processes. Therefore, it is important that more research activity be devoted to the investigation of the effects of age on these processes as this may be where mechanism(s) of action for the age-related decline in protein degradation lies. PMID- 11908072 TI - Protein degradation in Alzheimer's disease and aging of the brain. PMID- 11908073 TI - Protein degradation in human disease. PMID- 11908074 TI - The 26S proteasome. PMID- 11908075 TI - Cultural competence and health care: Japanese, Korean, and Indian patients in the United States. AB - Cultural competence requires sensitivity to the diverse ethnic, religious, and cultural expectations of patients in our health care system. In the increasingly multicultural world of the city hospital, patients will benefit from increased cultural competency on the part of health care providers. This study interviews Japanese, Korean, and Indian immigrants to the United States, showing that these individuals hold vastly different expectations concerning: 1) when to seek medical assistance; 2) the role of the doctor in the community; 3) the role of the patient and the patient's family in conversations with the medical specialist; 4) the roles of doctors versus nurses; 5) issues of privacy and disclosure to patient and family; 6) organ donation; and 7) end-of-life care. The paper concludes with immigrants' views on what would make their medical experience in the United States more comfortable, and hence, potentially more beneficial to their mental and physical health. PMID- 11908076 TI - Meeting folks where they are: collecting data from ethnic groups in the community. AB - Since the Nazi medical experiments in Europe and the Tuskegee Syphilis Study in the United States, members of ethnic groups have been reluctant to participate in research. However, a National Institutes of Health policy mandates that researchers develop and implement strategies to insure the inclusion of women and minorities as subjects in clinical investigations. The purpose of this paper is to discuss methods found to be effective in recruiting and retaining members of ethnic groups as participants in three research projects. The three studies were: (a) Gambling Behaviors in African American Elders and Perceived Effects on Health, (b) Adolescent Risk Behavior, Self Esteem, and Social Influence: Comparison of Dominican Youth in Their Homeland and in the United States, and (c) A Multicultural Study of Support for Alzheimer's Caregivers. PMID- 11908077 TI - American nursing students experience shock during a short-term international program. AB - When individuals plan to travel internationally, they frequently assume that they will have an enjoyable and memorable experience. But for some, the effects of culture shock may negatively impact their travels and memories. The purpose of this study was to describe culture shock as reported by student nurses who took part in an international short-term program. A phenomenological approach was utilized to elicit the essence of meaning attached to the experience. Eight student nurses in an upper Midwestern university, participated in this international experience. It was concluded that all of the student nurses experienced culture shock to a varying degree and they had varying perceptions of their experiences. PMID- 11908078 TI - A fast annealing evolutionary algorithm for global optimization. AB - By combining the aspect of population in genetic algorithms (GAs) and the simulated annealing algorithm (SAA), a novel algorithm, called fast annealing evolutionary algorithm (FAEA), is proposed. The algorithm is similar to the annealing evolutionary algorithm (AEA), and a very fast annealing technique is adopted for the annealing procedure. By an application of the algorithm to the optimization of test functions and a comparison of the algorithm with other stochastic optimization methods, it is shown that the algorithm is a highly efficient optimization method. It was also applied in optimization of Lennard Jones clusters and compared with other methods in this study. The results indicate that the algorithm is a good tool for the energy minimization problem. PMID- 11908079 TI - Studies on 4,7-di-substitution effects of one ligand in [Ru(phen)3]2+ with DFT method. AB - Studies on the complex [Ru(phen)3]2+ (phen = 1,10-phenanthroline) and its derivatives with 4,7-di-substitution on one ligand(phen) were carried out using the DFT method at the B3LYP/LanL2DZ level of theory. The trends in the substituent effects caused by the electron-pushing group (OH) and the electron withdrawing group (F), on the electronic structures and the related properties, for example, the energies and the components of some frontier molecular orbitals, the spectroscopy properties, and the net charge populations of some main atoms of the complexes, etc., have been investigated. The computational results show that the substituents have some interesting effects on the electronic structures and the related properties of the complexes. First, according to the analysis of components of LUMO of the complexes, the electron-withdrawing group (F) can activate the main ligand (the substituted ligand, i.e., 2R-phen) and passivate the coligands, on the contrary, the electron-pushing group (OH) can activate the coligands and passivate the main ligand in the first electronic excited states of complexes. Second, both the electron-pushing group (OH) and the electron withdrawing group (F) can cause a red shift in the electronic ground bands. Third, the characteristics of the atomic net charge populations on the main ligand can also be analyzed in detail by means of a schematic map expressed by several series of arrowheads based on the law of polarity alternation and the idea of polarity interference. The most negative charges are populated on N1, the next most net negative charges are populated on C3 among the skeleton atoms for the three complexes, etc. The computational results can be better used to explain some experimental phenomena and trends. PMID- 11908080 TI - Ab initio and DFT studies on van der Waals trimers: the OCS.(CO2)2 complexes. AB - Ab initio calculations [MP2, MP4SDTQ, and QCISD(T)] using different basis sets [6 31G(d,p), cc-pVXZ (X = D, T, Q), and aug-cc-pVDZ] and density functional theory [B3LYP/6-31G(d,p)] calculations were carried out to study the OCS.(CO2)2 van der Waals trimer. The DFT has proved inappropriate to the study of this type of systems where the dispersion forces are expected to play a relevant role. Three minima isomers (two noncyclic and one cyclic) were located and characterized. The most stable isomer exhibits a noncyclic barrel-like structure whose bond lengths, angles, rotational constants, and dipole moment agree quite well with the corresponding experimental values of the only structure observed in recent microwave spectroscopic studies. The energetic proximity of the three isomers, with stabilization energies of 1442, 1371, and 1307 cm-1, respectively, at the CBS-MP2/cc-pVXZ (X = D, T, Q) level, strongly suggests that the two unobserved structures should also be detected as in the case of the (CO2)3 trimer where both noncyclic and cyclic isomers have been reported to exist. The many-body symmetry adapted perturbation theory is employed to analyze the nature of the interactions leading to the formation of the different structures. The three-body contributions are small and stabilizing for the two most stable structures and almost negligible for the cyclic isomer. PMID- 11908081 TI - Crystal structure predictions using five space groups with two independent molecules. The case of small organic acids. AB - Crystal structure generations with two independent molecules have been performed for a series of carboxylic acids, using a slightly modified version of the OPLS force field. It was found that in this way the experimental structures with one independent molecule were produced as special cases, except for the molecules with four or more internal degrees of freedom. This work shows that a search with two independent molecules in only five space groups, although costly in computer power, can automatically also find structures with one independent molecule in many supergroups. Considering the observed abundances of structural classes, such a search should cover more than 95% of the possible homomolecular crystal structures. PMID- 11908082 TI - New method for parallel computation of Hessian matrix of conformational energy function in internal coordinates. AB - A new algorithm for parallel calculation of the second derivatives (Hessian) of the conformational energy function of biomolecules in internal coordinates is proposed. The basic scheme of this algorithm is the division of the entire calculation of the Hessian matrix (called "task") into subtasks and the optimization of the assignment of processors to each subtask by considering both the load balancing and reduction of the communication cost. A genetic algorithm is used for this optimization considering the dependencies between subtasks. We applied this method to a glutaminyl transfer RNA (Gln-tRNA) molecule for which the scalability of our previously developed parallel algorithm was significantly decreased when the large number of processors was used. The speedup for the calculation was 32.6 times with 60 processors, which is considerably better than the speedup for our previously reported parallel algorithm. The elapsed time for the calculation of subtasks, data sending, and data receiving was analyzed, and the effect of the optimization using the genetic algorithm is discussed. PMID- 11908083 TI - Calibration of force-field dependency in free energy landscapes of peptide conformations by quantum chemical calculations. AB - The free energy landscapes of peptide conformations were calibrated by ab initio quantum chemical calculations, after the enhanced conformational diversity search using the multicanonical molecular dynamics simulations. Three different potentials of mean force for an isolated dipeptide were individually obtained by the multicanonical molecular dynamics simulations using the conventional force fields, AMBER parm94, AMBER parm96, and CHARMm22. Each potential of mean force was then calibrated based upon the umbrella sampling algorithm from the adiabatic energy map that was calculated separately by the ab initio molecular orbital method, and all of the calibrated potentials of mean force coincided well. The calibration method was also applied to the simulations of a peptide dimer in explicit water models, and it was shown that the calibrated free energy landscapes did not depend on the force field used in the classical simulations, as far as the conformational space was sampled well. The current calibration method fuses the classical free energy calculation with the quantum chemical calculation, and it should generally make simulations for biomolecular systems much more reliable when combining with enhanced conformational sampling. PMID- 11908084 TI - Rapidly converging lattice sums for nonelectrostatic interactions. AB - The relative energies of one-, two-, and three-dimensional Bravais lattice Lennard-Jones particles can be calculated by lattice sums. The expression of lattice sums over a Lennard-Jones potential can be manipulated into a form that converges rapidly. A formalism capable of calculating the lattice potential at arbitrary points of a completely general lattice has been developed. This method provides an alternative way to calculate the relative energies from the surface and the interior bulk sites of many chemical systems. The method is illustrated with application to hcp and fcc Lennard-Jonesium, both for the relative binding energy and for calculating the potential along the geometric diffusion pathway between tetrahedral and octahedral interstitial sites. Diffusion from the tetrahedral site to the octahedral site experiences a barrier of 752.600 in units of 4 epsilon. The reverse pathway experiences a barrier of 1035.614 in units of 4 epsilon. PMID- 11908085 TI - New space warping method for the simulation of large-scale macromolecular conformational changes. AB - A space warping method, facilitating the modeling of large-scale conformational changes in mesoscopic systems, is presented. The method uses a set of "global (or collective) coordinates" that capture overall behavior, in conjunction with the set of atomic coordinates. Application of the space warping method to energy minimization is discussed. Several simulations where the method is used to determine the energy minimizing structures of simple central force systems are analyzed. Comparing the results and behavior of the space warping method to simulations involving atomic coordinates only, it is found that the space warping method scales better with system size and also finds lower minima when the potential energy surface has multiple minima. It is shown that the transformation of [Ala16]+ in vacuo from linear to globular is captured efficiently using the space warping method. PMID- 11908086 TI - Accurate prediction of proton chemical shifts. II. Peptide analogues. AB - Proton chemical shifts of eight cyclic amide molecules were measured in DMSO and D2O solutions. The magnetic shieldings of the corresponding aliphatic, aromatic, and amide protons were calculated by Hartree-Fock and DFT, using the 6-311G**, 6 311++G**, and TZVP basis sets. For aliphatic protons, all of these methods reproduce the experimental values in DMSO solutions excellently after linear regression. The Hartree-Fock method tends to give slightly better agreement than DFT. The best performance is given by the HF/6-311G** method, with an rms deviation of 0.068 ppm. The deviations from experimental chemical shifts in D2O solutions are only slightly larger than those in DMSO solutions. This suggests that we can use the calculated gas phase proton chemical shifts directly to predict experimental data in various solvents, including water. For amide protons, which exchange with water and form hydrogen bonds with DMSO, only modest agreement is obtained, as expected. The present studies confirm that the GIAO approach can reach high accuracy for the relative chemical shifts of aliphatic and aromatic protons at a low cost. Such calculations may provide constraints for the conformational analysis of proteins and other macromolecules. PMID- 11908087 TI - Improved semiempirical heats of formation through the use of bond and group equivalents. AB - Deficiencies in energetics obtained using the common semiempirical methods, AM1, PM3, and MNDO, may partly be traced to the use of pseudoatomic equivalents for conversion of molecular energies to heats of formation at 298 K. We present an alternative scheme based on the use of bond and group equivalents. Values for the 61 bond and group equivalents necessary for treatment of molecules containing the common organic elements, hydrogen, carbon, nitrogen, and oxygen have been derived. For a set of 583 neutral, closed-shell molecules mean absolute errors in AM1, PM3, and MNDO heats of formation are reduced from 6.6, 4.2, and 8.2 kcal/mol to 2.3, 2.2, and 3.0 kcal/mol, respectively. Several systematic problems are overcome in the present scheme including relative stabilities of branched hydrocarbons, energetics of conjugated systems, heats of formation of long chain hydrocarbons, and enthalpies of molecules containing multiple heteroatoms. Although the approach is restricted to molecules with well-defined functional groups, the equivalents are easy to incorporate and are chemically relevant. This revised procedure allows semiempirical methods to be used for far more reliable evaluations of heats of reactions. Estimates are made of the errors inherent in these semiempirical formalisms, arising from integral approximations and the neglect of explicit treatment of electron correlation effects, while excluding those from inadequate parameterization. PMID- 11908088 TI - Monte Carlo sampling algorithm for searching a scale-transformed energy space of polypeptides. AB - A Monte Carlo sampling algorithm for searching a scale-transformed conformational energy space of polypeptides is presented. This algorithm is based on the assumption that energy barriers can be overcome by a uniform sampling of the logarithmically transformed energy space. This algorithm is tested with Met enkephalin. For comparison, the entropy sampling Monte Carlo (ESMC) simulation is performed. First, the global minimum is easily found by the optimization of a scale-transformed energy space. With a new Monte Carlo sampling, energy barriers of 3000 kcal/mol are frequently overcome, and low-energy conformations are sampled more efficiently than with ESMC simulations. Several thermodynamic quantities are calculated with good accuracy. PMID- 11908089 TI - [Leptin--a hormone with many functions]. PMID- 11908090 TI - [Matrix metalloproteinases and their role in cancer progression]. PMID- 11908091 TI - [The role of cathepsin D in progression of neoplasms]. PMID- 11908092 TI - [Pseudouridine synthases--enzymes introducing the most abundant modified nucleoside in nucleic acids--pseudouridine]. PMID- 11908093 TI - [Genetic information under control--role of eukaryotic topoisomerase I]. PMID- 11908094 TI - [From differential gene expression to cDNA clone--review of methods for identification of genes with a variable level of transcription]. PMID- 11908095 TI - [Mechanism, regulation and role of bacterial bioluminescence]. PMID- 11908096 TI - [Using epidural anesthesia in patients with acute pancreatitis--prospective study of 121 patients]. AB - According to a previous study, an excellent level of analgesia can be expected when using epidural anaesthesia in patients with acute pancreatitis. In the present investigation, the effectiveness and safety of epidural anaesthesia is demonstrated in a large group of patients with severe acute pancreatitis, who were admitted to an intensive care unit. Epidural anaesthesia alone produced excellent analgesia on 1,083 of 1,496 observation days (72%) without the systemic use of other analgesic substances. Even in patients with marginal cardiovascular stability, epidural injection of local anaesthetic solution was tolerated well. Only 8% of all local anaesthetic injections were associated with a haemodynamic reaction that required pharmacological intervention. There was no case of a septic or neurological complication of epidural anaesthesia. Initially elevated serum amylase and lipase were normalized after 17.4 days (minimum one day, maximum 19 days). Surgical intervention was necessary for 36 patients, with a total of 64 surgeries having to be performed, including cholecystectomy. Sixteen patients required artificial ventilation for an average time of 12.3 days (minimum two days, maximum 48 days). Lethality was 2.5% (three patients), with all three patients suffering from an acute stage III pancreatitis. The average duration of ICU treatment was 12.4 days (minimum two days, maximum 101 days). PMID- 11908097 TI - [Etiology and sequelae of perioperative accidental hypothermia]. AB - Accidental hypothermia is a frequent event during the perioperative period. Recent studies revealed a drop in core temperature of over 2 degrees C in more than 50% of all patients undergoing an operation. This drop in core temperature seems to be primarily due to the following factors. Anaesthesia prevents behavioural adaptations to changes in ambient temperature. Simultaneously, autonomic mechanisms of temperature control are suppressed by general as well as by neuraxial anaesthesia. The interthreshold range between core temperatures that trigger responses to warmth and to cold increases up to 20-fold. This is primarily due to a decrease in the cold response threshold. As a result, body core temperature of anaesthetized patients is primarily determined by the much lower temperature of the environment. On one hand, decreases in body temperatures may exert organ protective effects under certain conditions, e.g., by increasing ischemic tolerance. On the other hand, there is accumulating evidence that accidental perioperative hypothermia may also adversely affect organ function and outcome. For example, unfavourable effects of perioperative hypothermia on the immune defence, on the function of the coagulation system, on cardiovascular performance, as well as on postoperative recovery have been reported. Consequently, measures should be taken to actively control the perioperative heat balance of patients. PMID- 11908098 TI - [Accidental hypothermia--a challenge for rescue service and intensive care]. AB - Accidental hypothermia is a rare clinical picture with different causes. Specific features are shown by patients who have accidents in water, due to rapid cooling. The SARRRAH project (Search and Rescue, Resuscitation and Rewarming in Accidental Hypothermia) was launched to secure fast and professional medical care right up to rewarming by extracorporal circulation. The University of Rostock takes part in this project. Based on the course of accidental hypothermia in fifteen patients, the authors report on the treatment of this life-threatening situation with special regard to the use of extracorporal circulation and present their first results. The core temperature of these patients lay between 16.0 and 34.0 degrees C. Eight of the patients had cardiac arrest at the scene of the accident. Seven of the patients with cardiac arrest were treated with extracorporal circulation in addition to cardiopulmonary resuscitation, which was started pre clinically and continued in hospital. In one patient, extracorporal circulation was used at an initial temperature of 25.4 degrees C without previous cardiopulmonary resuscitation. Six of these fifteen patients with accidental hypothermia died. Five of the non-survivors belonged to the group of eight patients who were rewarmed by extracorporal circulation. With one exception, they also had the lowest core temperatures. Only a homogeneous and up-to-date documentation will allow further conclusions to be made for improving the concept of therapy. PMID- 11908099 TI - Two aged care facilities to close. PMID- 11908100 TI - Hospital offers holistic health care. PMID- 11908101 TI - Fewer public hospital beds. PMID- 11908102 TI - Hungarian nurses win compensation. PMID- 11908103 TI - African HIV/AIDS causes nurse exodus. PMID- 11908104 TI - Important news for Vietnam nurses. The campaign continues. PMID- 11908106 TI - A collaborative approach to promoting safe maternity care. PMID- 11908105 TI - Defending wages and conditions. PMID- 11908107 TI - Caesarean rates continue to climb. PMID- 11908108 TI - Pay central to nursing shortages. PMID- 11908109 TI - Mental health beds in decline. PMID- 11908110 TI - Midwives lose indemnity insurance. PMID- 11908111 TI - Sexually transmissible infections. PMID- 11908112 TI - NT industrial action update. PMID- 11908113 TI - WA Nurses Board report shows aged care in crisis. PMID- 11908114 TI - Globalisation advances on public health. PMID- 11908115 TI - Improving access to health care. PMID- 11908116 TI - Aged care: compassion and commitment. PMID- 11908117 TI - Trial reaches halfway stage. Another step nearer to nurse practitioner in the ACT. PMID- 11908118 TI - Occupational exposure: there for your protection. PMID- 11908120 TI - Improving lupus knowledge. PMID- 11908119 TI - Medication learning packages for new graduates. PMID- 11908121 TI - Course offers insight into Aboriginal health. PMID- 11908122 TI - Treating the Jehovah's Witness patient. PMID- 11908123 TI - Nurses speaking out about what they do. Interview by Heather Witham. PMID- 11908124 TI - Not doing nothing. PMID- 11908125 TI - NSW nurses ask: what's a nurse worth? PMID- 11908126 TI - Victorian nurses lift bans as a sign of goodwill. PMID- 11908127 TI - Hospital deaths prompt calls for reform. PMID- 11908128 TI - Multiple sclerosis and the role of the MS nurse consultant. PMID- 11908129 TI - [Topical tacrolimus in dermatology: what can one expect?]. PMID- 11908130 TI - [Consideration of African albinos]. PMID- 11908131 TI - [Anti-endothelial cell antibodies in chronic leg ulcer: prevalence and significance]. AB - BACKGROUND: Anti-endothelial cell antibodies are detected in auto-immune vasculitis and connective tissue diseases such as scleroderma, with possible pathogenic involvement. We looked for these antibodies in patients presenting chronic ulcers. PATIENTS AND METHODS: Eighty patients were tested: 35 presented vascular ulcers (27 of venous origin and 8 arterial). A control group of 14 patients with connective tissue disease and vasculitis was formed, and a third group included 31 patients without vascular or immune disease. Their sera were tested by ELISA technique on EAhy 926 cell (obtained from hybridization of human endothelial and line A 549/8 cells). RESULTS: Blood tests were positive in 37.5 p. 100 of patients. Antibodies were present in 48.5 p. 100 of patients with leg ulcers (of which 59 p. 100 (16/27) venous ulcers). Fifty-seven per cent of the autoimmune group and 12 p. 100 of the third group were positive. Association with anti-phospholipid antibodies was observed twice in leg ulcers. The level of antibodies was higher in patients with leg ulcers. COMMENTS: The presence of high levels of anti-endothelial cells antibodies in 48.5 p. 100 of patients presenting leg ulcers shows that these antibodies are not specific to autoimmune diseases. Since they are involved in coagulation, inflammation (enhancing of adhesion molecules and tissue lysis) and apoptosis of endothelial cells, their significance in chronic ulcers, both as a consequence and/or possible cofactor merits discussion. PMID- 11908132 TI - [Lyell syndrome in Senegal: responsibility of thiacetazone]. AB - INTRODUCTION: Toxic epidermal necrolysis is a severe disease often leading to death or to mucosal, particularly ocular, after effects. The principle drugs responsible are antibacterial sulfonamides, anti-epileptics, non-steroid anti inflammatories, allopurinol and chlormezanone. We report a series of 38 cases of toxic epidermal necrolysis, observed in Dakar, imputable to thiacetazone and lethal in 60 percent of cases. PATIENTS AND METHODS: Our study was retrospective. Diagnosis of toxic epidermal necrolysis was made in patients presenting more than 30 p. 100 of the epidermis of their total body surface stripped off, multi orificial mucosal damage and epidermal necrosis revealed on histological examination. Drug imputability was established on classical criteria. Treatment was composed of reanimation and antibiotics. RESULTS: Among the 38 cases of toxic epidermal necrolysis counted, 24 were imputable to thiacetazone. All the patients presented typical clinical features, confirmed histologically. Evolution was lethal in 60 p. 100 of cases. The causes of death were frequently hypovolemic shock during the first week and septic shock during the second. The deceased were generally aged over 50, had more than 50 p. 100 of total epidermis stripped off, presented evolving tuberculosis at the time of the accident and HIV infection at the AIDS stage. After effects were vaginal synechia and 2 cases of blindness. COMMENTS: Our series is exceptional in that a) the drug responsible: thiacetazone, an economic tuberculostatic of minor efficacy, was systematically introduced after 2 months of intensive treatment with 4 major anti-tuberculosis agents; b) the 60 percent mortality rate, two-fold greater that that usually observed. Other than the known elements of poor prognosis in our patients, the treatment conditions of this dermatological emergency explain this high rate of mortality. PMID- 11908133 TI - [Survival after cutaneous metastasis: a study of 200 cases]. AB - BACKGROUND: Cutaneous metastatic disease is uncommon and the outcome after cutaneous metastasis has rarely been thoroughly studied. The objective of this work was to study the survival after diagnosis of cutaneous metastasis in a large series of patients and to evaluate survival according to the type of cancer. PATIENTS AND METHODS: This retrospective study was conducted out in the Laboratoire d'Histo-pathologie Cutanee of Strasbourg. Between 1950 to 1996, 228 patients with cutaneous metastasis were diagnosed on the basis of typical histopathology, confirmed by two dermatopathologists. We excluded lymphoma or leukaemia with secondary skin involvement. Medical and demographic data were collected from hospital data, and the "Registre du Cancer du Bas-Rhin". The type of neoplasm, the time of diagnosis of primary cancer and the time of death (or survival at 12/31/1996) was established in 200 patients, 99 men and 101 women with a mean age 62.4 +/- 13 years. We found 64 cases of breast carcinoma, 36 cases of lung carcinoma, 31 cases of melanoma and 69 cases of other cancers. Long term actuarial survival after cutaneous metastasis was calculated using by the Kaplan-Meier method. RESULTS: The median survival after cutaneous metastasis was 6.5 months (mean 22.8 +/- 43.8 months). The mortality rate was 13 p. 100 at 1 month, 48 p. 100 at 6 months and 64.5 p. 100 at 12 months. Median survival was calculated according to the primary neoplasm: breast carcinoma: 13.8 months, melanoma: 13.5 months, lung carcinoma: 2.9 months (36 cases). The outcome of patients with cutaneous metastasis of lung carcinoma was worse than those with melanoma (p < 10(-4)) and breast cancer (p < 10(-4)). Survival after cutaneous metastasis of other cancers could not be compared because of the small size of the subgroups: median survival after cutaneous metastasis of non cutaneous squamous cell carcinoma of the head and neck: 8.8 months (5 cases), cutaneous squamous cell carcinoma: 6.5 months (12 cases), carcinoma of oesophagus: 4.7 months (2 cases), colo-rectal cancer: 4.4 months (9 cases), pancreatic cancer: 3.3 months (2 cases), stomach cancer: 1.2 months (7 cases) and liver and gall bladder carcinoma: < 1 month (3 cases). Survival beyond 10 years was observed in 9 patients: 3 melanoma, 2 breast cancers, 2 prostatic carcinomas, 1 larynx carcinoma and 1 cutaneous squamous cell carcinoma. CONCLUSION: This is the first study in which the survival after the occurrence of skin metastasis was systematically analysed in a large series of patients. It shows that half of patients with cutaneous metastasis die within the first 6 months after the diagnosis. Those cases due to lung carcinoma have the poorest prognosis. PMID- 11908134 TI - [Griseofulvin]. AB - Griseofulvin is a metabolic product of Penicillium spp. It was the first available oral agent for the treatment of dermatophytoses and has now been used for more than forty years. Griseofulvin is fongistatic, the exact mechanism in witch it inhibits the growth of dermatophytes is doubtful. Several ways are invoked: inhibition of fungal cell mitosis and nuclear acid synthesis, probable interference with the function of microtubules. Griseofulvin is poorly absorbed from the gastrointestinal tract. Absorption is enhanced by administration with fatty meal. Peak plasma occurs four hours after oral administration. Griseofulvin is detected in the outer layer of the stratum corneum soon after it is ingested, it is diffused from the extracellular fluid and sweat. There is no information regarding the mechanism by witch the drug is delivered to nails and hair. Deposition in the newly formed cells could be the major factor. Griseofulvin has also anti-inflammatory properties and some direct vasodilatory effects when it is used in high doses. It is metabolised by the liver microsomial enzyme system and excreted in the urine. The half-life is 9 to 21 hours. Griseofulvine has been used in the therapy of dermatophyte onychomycosis, treatment periods from 6 to 18 months were necessary with disappointing results and numerous relapses. Newer oral antifungal agents are now preferred especially in toenail infections. For many authors griseofulvin is still the treatment of choice of tinea capitis. Doses are 15-20 mg/kg/d for 6 to 8 weeks in children with the microsized form. Clinical response rates have been reported between 80 and 90 p. 100 in controlled studies. Griseofulvin is well-tolerated particulary in children. More frequent side effects are minor: headaches, gastrointestinal reactions and cutaneous eruptions. The major drug interactions has been noted with phenobarbital, anticoagulants and oral contraceptives. PMID- 11908135 TI - [Mouth ulcers in patients receiving tacrolimus]. AB - INTRODUCTION: The buccal side effects of immunodepressors are well defined with cyclosporine and certain antimitotic agents. We report a case of buccal ulcerations in a patient treated with a new immunosuppressive macrolide: tacrolimus (Prograf). OBSERVATION: A 53 year-old woman presenting a severe cardio myopathy, underwent heart transplantation in May 1997. Tacrolimus was introduced in October 1997 after 3 episodes of acute reject. Eight months after tacrolimus, painful apthoid buccal ulcerations appeared. Biopsy of the buccal mucosa and other biological examinations revealed no particular etiology. Since tacrolimus could not be stopped, treatment with thalidomide was initiated. It was suspended on two occasions due to adverse events. The buccal ulcerations relapsed rapidly. The intrinsic imputability of tacrolimus in the occurrence of these lesions was noted "l2" ("plausible"). DISCUSSION: Several arguments suggest that these buccal ulcerations may result from the toxicity of tacrolimus: 1) absence of past history of apthae; 2) anatomo-clinical aspect of the lesion differing from that of common apthae, but similar to the ulcerations observed with nicorandil; 3) delay in occurrence of analogous ulcerations compared with that observed with methotrexate or nicorandil; 4) absence of another etiology; 5) relapse of ulcerations on two occasions after suspension of thalidomide, whilst tacrolimus was continued. PMID- 11908136 TI - [Schopf-Schulz-Passarge syndrome: 2 cases]. AB - BACKGROUND: Schopf-Schulz-Passarge's syndrome is a rare autosomal recessive genodermatosis associating hypodontia, palmoplantar keratoderma, cysts of the eyelid margins, onychodysplasia and hypotrichosis. We report two new cases. CASE REPORT: Case no. 1: A 49 year-old woman complained of erosive and fissured palmoplantar keratoderma. Nails were fragile and dystrophic. Permanent teeth were absent. She also had many small cysts of the eyelid margins and a middle hypotrichosis. There was no consanguinity between her parents. Case no. 2: A 56 year-old man was seen for red, scaly and well marked palmoplantar keratoderma. Permanent teeth were absent. He had a hair loss since the age of 30. Nails were hypoplastic and there were many small cysts of the eyelids. Biopsy of one of the cysts showed a follicular cyst associated with sweat duct dystrophy. Schopf Schulz-Passarge's syndrome was diagnosed in these 2 patients. There was no evidence of associated cutaneous tumors. DISCUSSION: Differential diagnosis of Schopf-Schulz-Passarge syndrome include other genodermatoses comprising palmoplantar keratoderma and dental abnormalities. Benign or malignant tumors are frequently associated: eccrine poromas, eccrine syringofibroadenomas, follicular tumors, basal cell and squamous cell carcinomas. Tumors usually appear after the age of 60. Regular follow-up and biopsy of the suspect lesions are necessary. PMID- 11908137 TI - [Hamartoma oligemicus]. AB - BACKGROUND: Naevus oligemicus is a rare affection caused by selective vasoconstriction of the deeper vessels ("thermal" skin blood flow), with respect of the superficial vascular plexus ("nutrient" blood flow). We report the fourth case. CASE REPORT: A 45 year-old man presented erythematocyanotic lesions with sensation of coolness on the abdomen and the thighs. These lesions had first been noted by the patient 3 years earlier. Physical examination and laboratory investigations were normal. Histology of a lesion revealed a non-specific superficial dermatitis. Result of skin surface thermometry of both areas showed that the surface temperature of the involved skin was up to 2 degrees C lower than the surrounding control skin. DISCUSSION: We don't know why there is a selective vasoconstriction of the deeper vessels and not of the superficial vascular plexus. Naevus oligemicus belongs to the pharmacological naevus group because of the absence of any histologic vascular abnormality. The functional abnormality has not been identified. This could be explained either by increased sympathetic vasoconstrictor tone in the autonomic sympathetic vasoconstrictor nerves supplying the deeper thermoregulatory vasculature in the affected skin, or by increased sensitivity of these vessels to vasoconstrictor catecholamines. PMID- 11908138 TI - [Intravascular papillary endothelial hyperplasia: multiple lesions appearing in the course of treatment with interferon beta]. AB - INTRODUCTION: Intravascular papillary endothelial hyperplasia is a benign and rare vascular lesion. We report a case of multiple vascular lesions of the hand following 3 months treatment with beta-interferon injections for multiple sclerosis. OBSERVATION: A 50 year-old man had multiple vascular nodules of the hands. He was treated with beta interferon injections for multiple sclerosis for 3 months. Histology showed typical changes of intravascular papillary endothelial hyperplasia: papillary endothelial proliferation in a dilated cavity associated with thrombosis. DISCUSSION: Intravascular papillary endothelial hyperplasia is a benign and rare vascular lesion usually presenting as a simple nodule. It may be painful. Diagnosis is histologic, characterized by papillary endothelial proliferation associated with a thrombus within a vessel. It may be confused with hemangiosarcoma. Treatment is surgical and recurrence after treatment is rare. Intravascular papillary endothelial hyperplasia is generally considered as an unusual form of thrombus organization. Intravascular papillary endothelial hyperplasia is divided into two groups: a pure form occurring within a dilated vessel and a mixed form appearing in benign vascular lesions. The originality of this case is the rarity and the multiplicity of the lesions. The possible pathogenesis of interferon-induced cutaneous vascular lesions is discussed. PMID- 11908139 TI - [Panniculitis and macrophage activation syndrome in a child with lupus erythematosus]. AB - INTRODUCTION: Panniculitis is rarely presented in the course of systemic lupus erythematosus. When it occurs, it is mainly related to lupus profundus. However, when panniculitis is associated with a reactive hemophagocytic syndrome, panniculitis could be linked to this hemophagocytosis reaction. CASE REPORT: We report a case of an 11-year-old girl treated for several years for systemic lupus erythematosus, who simultaneously presented panniculitis and an hemophagocytic syndrome. The reality of both diagnoses was based on the analysis of biological and histological data. Therapy with immunosuppressive drugs led to relief of the symptoms. DISCUSSION: We emphasize our discussion on the pathophysiology of lupus profundus and hemophagocytosis with regards to the role of cytokines and circulating immune complexes in both diseases. They may enhance the hypodermal necrosis observed in lupus profundus and induce macrophage cell dysregulation through cytotoxic cells known in hemophagocytosis. The immunomodulative action of immunosuppressive therapy with inhibition of cells involved in immune response, may explain its efficacy. PMID- 11908140 TI - [Familial hyperchylomicronemia with a new mutation of the lipoprotein lipase gene]. AB - INTRODUCTION: Familial hyperchylomicronemia is a rare autosomal recessive disease caused by lipoprotein lipase deficiency. CASE-REPORT: A nine month-old girl presented with eruptive xanthomas revealing a familial hyperchylomicronemia. No lipoprotein lipase activity was found. DNA analysis revealed a novel homozygous non-sense mutation of the lipoprotein lipase gene at the codon 288. The parents were heterozygous carriers. DISCUSSION: Familial hyperchylomicronemia usually presents with eruptiva xanthomas, abdominal pain, pancreatic manifestation and lipemia retinalis. Papulo-nodular xanthomas occur more frequently in children as in our case. Eighty lipoprotein lipase gene mutations have been recorded to date. The gene locates on chromosome 8. Only 9 non-sense mutations have been described which lead to a truncated protein. In our case, no enzymatic activity was detected probably due to an absence of secretion of the enzyme, even though catalytic activity persisted. The homozygous carrier status leads to hyperchylomicronemia whereas the heterozygote status may lead to mixed hyperlipidemia with an increased risk of atherosclerosis. The screening of lipoprotein lipase gene mutations should be carried out in all families with hyperchylomicronemia, regardless of the presence or absence of xanthomas. PMID- 11908141 TI - [Atopic dermatitis: survey of the quality of life of patients and their care]. PMID- 11908142 TI - [Thromboangiitis obliterans and arsenic poisoning]. PMID- 11908143 TI - [Evaluation of atopic dermatitis in routine general practice in Algeria]. PMID- 11908144 TI - [Flat angioma and idiopathic facial neuralgia. Association or coincidence?]. PMID- 11908145 TI - [Drug hypersensitivity syndrome rapidly resolving after human immunoglobulin infusion]. PMID- 11908146 TI - [Exophytic tumor of the scalp]. PMID- 11908148 TI - [Hypersensitivity to neomycin]. PMID- 11908147 TI - [Chronic evolution of papules]. PMID- 11908149 TI - [Single erythroplastic vulvar lesion in a 70-year-old woman: Bowen disease]. PMID- 11908150 TI - [Etiologic agents in contact urticaria]. PMID- 11908151 TI - [Basal cell carcinoma of a light exposed area in a dark skinned African woman]. PMID- 11908152 TI - [Management of mucocutaneous herpes in the immunocompetent patient]. PMID- 11908153 TI - [Solupred in pediatrics]. PMID- 11908154 TI - [Toxocariasis]. PMID- 11908155 TI - [Baboon syndrome: frontline new for the backside]. PMID- 11908156 TI - [Francois Henri Hallopeau (1842-1919)]. PMID- 11908157 TI - [Juvenile xanthogranuloma]. PMID- 11908158 TI - [Reactivation of herpesvirus 2 genital infection in seropositive asymptomatic subjects]. PMID- 11908159 TI - [Is vitamin C deficiency a current health problem?]. PMID- 11908160 TI - [Medical information for the patient]. PMID- 11908161 TI - [Managing chronic wounds. A new rubrics for continuing medical training in the Annales de Dermatologie]. PMID- 11908163 TI - [Does subcutaneous cellular tissue exist?]. AB - BACKGROUND: The definition of the "subcutaneous cellular tissue" is still debated. METHODS: In order to establish the localisation and composition of this tissue, or, merely its existence, we interviewed French dermatologists and conducted a bibliographic study on the historical definitions of the so-called "subcutaneous cellular tissue". A questionnaire was sent to French professors of dermatology to assess their definition of the "subcutaneous cellular tissue". They were also asked to make a simple cartoon showing the anatomy of the skin and "subcutaneous cellular tissue". RESULTS: We obtained 37 answers which could be classified in three main categories: 1) "subcutaneous cellular tissue" and hypodermis are synonymous, 2) "subcutaneous cellular tissue" is an autonomous tissue which separates the hypodermis from the tissues below and 3) "subcutaneous cellular tissue" designates all structures located below the hypodermis. In an historic perspective, the "cellular system" was a macroscopic concept described in the eighteenth century as whitish fibrils delimitating "cells". It was renamed loose connective tissue in the twentieth century and thus "cellular" became obsolete. The definition of skin, and of "subcutaneous cellular tissue" in particular, has greatly changed over time. In the eighteenth century, only epidermis and dermis were considered as belonging to the skin, although some included the tela subcutanea. In the twentieth century, the "subcutaneous cellular tissue" is considered either as a part of the hypodermis, or as the hypodermis itself or as a tissue located between the hypodermis and the fascia. DISCUSSION: The difficulty in defining "subcutaneous cellular tissue" is the result of a French semantic problem. The "cellular tissue" became loose connective tissue. For French dermatologists, the skin is composed of epidermis, dermis and hypodermis and the hypodermis can therefore not be subcutaneous. In order to check whether an autonomous tissue could be evidenced under the skin, we conducted an histologic study, which is presented in a second article. PMID- 11908162 TI - [Managing chronic wounds. Knowledge and practice of nurses]. AB - BACKGROUND: Recommendations on chronic wound management have greatly evolved during the last years. Practice in the medical units of the Civil Hospital of Colmar (France) appears to controversy these recommendations. Since nurses have a central place in such cares, we conducted a management audit among those working in different departments, before organising an educational program. METHODS: One hundred and ten nurses (20 p. 100), randomly selected among the 546 concerned by chronic wounds working in the hospital were invited to participate in this audit composed of 23 questions. RESULTS: Participation rate was 72 p. 100. Response rates were higher than 80 p. 100 for 18 of 23 questions. The percentages below are those of the expressed answers. For 58 p. 100 of nurses executing the care after medical prescription, 31 p. 100 did so without medical advice and 11 p. 100 in collaboration with a doctor. Five and 14 p. 100 respectively knew all the classical clinical characteristics of venous and arterial leg ulcers. Some important risk factors for pressure sores were not well known, and only 15 p. 100 of nurses regularly used an evaluation scale of this risk. The massage-kneading of a stage I pressure ulcer was practiced by 49 p. 100 of nurses. Depending on the questions, 64 to 82 p. 100 of them routinely used antiseptics, and 82 p. 100 thought it was more important than compression for the healing of a venous leg ulcer. In more than one third of the answers, the dressing of a chronic wound was described as sterile. Finally, if an average knowledge of hydrocolloids was observed, there was little on alginates, foams and hydrogels, with a majority of nurses avoiding to answer these questions. COMMENTS: Although important is the management of chronic wounds, basic knowledge was insufficient to hope for the nurses' collaboration. Prevention policy of pressure sores was not optimal. There were discrepancies between local practices and actual knowledge. Knowledge of new dressings was inadequate, and there was a risk for limited clinical benefit. We believe the results would have been similar if the audit had conducted among the doctors. Hense, a vast educational program has been initiated. PMID- 11908164 TI - [Cellular subcutaneous tissue. Anatomic observations]. AB - INTRODUCTION: We showed in a companion paper that the definition of the French "subcutaneous cellular tissue" considerably varied from the 18th to the end of the 20th centuries and has not yet reached a consensus. To address the anatomic reality of this "subcutaneous cellular tissue", we investigated the anatomic structures underlying the fat tissue in normal human skin. METHODS: Sixty specimens were excised from the surface to the deep structures (bone, muscle, cartilage) on different body sites of 3 cadavers from the Institut d'Anatomie Normale de Strasbourg. Samples were paraffin-embedded, stained and analysed with a binocular microscope taking x 1 photographs. Specimens were also excised and fixed after subcutaneous injection of Indian ink, after mechanic tissue splitting and after performing artificial skin folds. RESULTS: The aspects of the deep parts of the skin greatly varied according to their anatomic localisation. Below the adipose tissue, we often found a lamellar fibrous layer which extended from the interlobular septa and contained horizontally distributed fat cells. No specific tissue below the hypodermis was observed. Artificial skin folds concerned either exclusively the dermis, when they were superficial or included the hypodermis, but no specific structure was apparent in the center of the fold. India ink diffused to the adipose tissue, mainly along the septa, but did not localise in a specific subcutaneous compartment. DISCUSSION: This study shows that the histologic aspects of the deep part of the skin depend mainly on the anatomic localisation. Skin is composed of epidermis, dermis and hypodermis and thus the hypodermis can not be considered as being "subcutaneous". A difficult to individualise, fibrous lamellar structure in continuity with the interlobular septa is often found under the fat lobules. This structure is a cleavage line, as is always the case with loose connective tissues, but belongs to the hypodermis (i.e. fat tissue). No specific tissue nor any virtual space was observed below the skin. Thus, the commonly used term "subcutaneous cellular tissue" is inappropriate. PMID- 11908165 TI - [Severe pruriginous acne in dialysed renal failure. Diagnostic difficulties and efficacy of isotretinoin]. AB - INTRODUCTION: The occurrence of acne in dialysed renal failure patients has rarely been reported and the clinical characteristics and therapeutic issues rarely studied in these patients. CASE REPORTS: Two men and two women, 33 to 56 years-old, with chronic renal failure and no past history of acne, developed severe acne under dialysis. The acne was excoriated in all cases and associated with prurigo-like lesions and intense pruritus, which made diagnosis difficult. Acne was profuse on the face and the trunk, but also on the neck (1 case) and the upper limbs (2 cases). No patient was taking acne-inducing substances. Various to therapies attempting to control pruritus were ineffective. However, anti-acne treatments (cyclines associated with local tretinoin in 1 case and oral isotreninoin in 3 cases) led to complete regression of the acne, pruritus and the prurigo-like lesions without relapse after a follow-up time of 4 months to 2 years. DISCUSSION: Pruritus is frequent during renal failure. However, the occurrence of unexplained acne has only rarely been reported. Our patients' clinical picture was original, characterized by the late development, under dialysis, of severe and pruriginous acne, the pathogenesis of which is unknown. Because of the clinical and therapeutic implications (impaired quality of life, pigmentation or scarring and remarkable efficacy of oral isotretinoin) this clinical picture merits more attention, and the modalities for the prescription of isotretinoin in this context should be defined. PMID- 11908166 TI - [Bowen disease treated with scanned pulsed high energy CO2 laser. Follow-up of 6 cases]. AB - INTRODUCTION: Cutaneous Bowen's disease is an intra-epidermal squamous cell carcinoma. PATIENTS AND METHOD: Ten cases of cutaneous Bowen's disease diagnosed among 8 patients were treated by scanned high energy carbon dioxide laser between November 1996, and March 1998. A biopsy was performed in all patients before treatment. RESULTS: The post-treatment follow-up extended from 1 to 4 years with an average follow-up of 2 years and 11 months. Only one patient, whose lesion was located on the auricle, presented a recurrence after one year. The remaining patients did not present any recurrence during their last control: six patients were followed for two years or more and one patient for one year. We demonstrate a histological and clinical correlation between the number of carbon dioxide laser passes before a clinical endpoint and the thickness of the epidermal carcinoma treated. DISCUSSION: This new treatment has comparable efficacy to other treatments. It can be applied to extensive lesions without sequelae except for the risk of residual hypopigmentation. PMID- 11908167 TI - [Survey in 2000: 3 cases]. AB - BACKGROUND: We report three new cases of patients presenting scurvy. In the year 2000 this rare disease still occurs in France. CASE REPORTS: The three patients, 2 men and a woman respectively 51, 50, and 73 years-old were alcoholics, and lived alone in difficult social conditions. Dietary survey indicated in the 3 cases inadequate vitamin C intake, and a regimen including solely bread, rice, pasta, and packet soup devoid of fresh vegetables and fruit. The cutaneous findings attributed to scurvy were: in the first patient, a woody inflammatory and painful oedema of the left leg associated with perifollicular petechial haemorrhages over the lower limbs, and hyperpigmentation of the facial skin with slate-gray spotty pigmentation of the tongue (pseudo-addisonian hyperpigmentation); in the second patient, an accentuation of a pre-existing acne becoming more inflammatory and extensive; and in the third patient, a diffuse petechial eruption on the abdomen and lower extremities. The diagnosis of scurvy was confirmed by low plasma ascorbic acid levels (< 6 mumol/l). All patients were treated with 1 to 2 g of oral ascorbic acid daily for 2 weeks resulting in rapid and dramatic response. DISCUSSION: Scurvy is a rare disease in industrialized nations. Its incidence is unknown because of absence of total census. Dietary vitamin C deficiency represents the main risk factor exposing for scurvy among adults, often alcoholics and living in social isolation. Cutaneous features supporting the diagnosis of scurvy are described in our observations. The recognition of these cutaneous abnormalities is important because their association can be misleading, and erroneously interpreted as a sign of systemic vasculitis, or connective tissue disease. The diagnosis of scurvy is confirmed by the measurement of plasma ascorbic acid levels. Treatment is simple and based on the administration of vitamin C, which results in dramatic improvement. PMID- 11908168 TI - [Pemphigoid and acquired hemophilia]. AB - INTRODUCTION: The association of bullous pemphigoid and acquired haemophilia is reported. CASE-REPORT: A 74 year-old man developed a bullous pemphigoid after decreasing corticotherapy, ecchymosis and haematomas revealing a high level of acquired anti-VIII antibodies (110 Bethesda UB units; TCA 98 s). Immunosuppressive treatment (cyclosporine, prednisone, azathioprine and bolus of cyclophosphamide) did not stop the disease. Perfusion of recombinant factor VIIa, human immunoglobulins and prednisone-azathioprine association permitted clinical and biological remission. DISCUSSION: Acquired haemophilia is idiopathic half the time. It can appear in autoimmmune diseases. Mortality is high. Only 4 cases of association with bullous pemphigoid have been reported in the literature. At the haemorrhagic phase, porcine factor VIII or more recombinant activated factor VII with human immunoglobulins are necessary. Immunosuppressive treatment is used to decrease production of anti-factor VIII antibodies. PMID- 11908169 TI - [Acute osteomyelitis. A rare erysipelas differential diagnosis]. AB - INTRODUCTION: Acute haematogenous osteomyelitis, whose clinical features may mimic erysipelas, is an uncommon disease in adults. OBSERVATION: A 56 year-old man was hospitalized for a suspicion of leg erysipelas. Oral and intravenous antibiotic therapy was inefficient. Magnetic Resonance Imaging (MRI) of the leg revealed osteomyelitis with subperiosteal abscess. Change of antibiotics and surgical debridement improved the patient's condition. DISCUSSION: Erysipela is a common disease which most often responds to anti-streptococcal therapy. Unfavourable evolution under antibiotherapy must lead to consider necrotizing fasciitis but also acute osteomyelitis. In these cases MRI is necessary. In our observation, the leg pain which preceded other signs of local inflammation, suggested the existence of primitive bone infection which further diffused in soft tissues, thus explaining the erysipelatoid aspect. PMID- 11908170 TI - [Cutaneous acanthamoebiasis in a lung transplant patient]. AB - BACKGROUND: Cutaneous acanthamoebiasis is a rare opportunistic infection in immunocompromised patients, particularly in HIV infected patients. We report a case of a primary cutaneous Acanthamoeba infection, in a patient with a double lung transplant not infected by the HIV. CASE REPORT: A 38 year-old white man, with a double lung transplant 3 years ago, developed a painful fibrinous ulceration of the right foot, with cellulitis. A few days later, two purplish nodules were observed, surrounded by an inflammatory reaction. Histologic examination revealed trophozoite and cyst forms. Therapy was begun with intravenous pentamidine and itraconazole along with topical ketoconazole and chlorhexidine, but was ineffective. Because of the renal toxicity of pentamidine, the patient was treated by dialysis. He died six months after diagnosis of Acanthamoeba infection. DISCUSSION: Acanthamoeba, a free-living amoeba is the causative organism of few human diseases. In immunocompromised hosts (particularly HIV infected), besides granulomatous amebic encephalitis, it can provoke some cutaneous lesions such as nodules, pustules, ulcerations... Skin biopsy is diagnostic: numerous amebic trophozoites and cysts are visualized. Recommended treatments are pentamidine, itraconazole and flucytosine. Due to the morbidity and mortality of acanthamoeba infection optimal therapy must be defined. PMID- 11908171 TI - [Androgenic alopecia revealing an androgen secreting ovarian tumor]. AB - INTRODUCTION: Androgen-producing tumors of the ovary are rare in postmenopausal women and are revealed by severe virilization. Leydig hilus cell tumors are the most frequent postmenopausal virilizing tumors. In this report, an unusual and rare cause of alopecia due to Leydig cell hyperplasia within the wall of a simple cyst and in the ovarian hilus is described. OBSERVATION: An 80 year-old woman complained of a 10-year history of severe androgenic alopecia associated with very mild facial hirsutism, without others signs of virilization. Hormonal blood levels showed markedly elevated testosterone. Computed tomographic scan of the adrenals and the ovaries revealed an enormous left ovarian cystic mass. Bilateral hystero-ophorectomy was performed. Histological examination demonstrated bilateral Leydig cell hyperplasia within the wall of the cyst and in the right ovarian hilus. Two months postoperative hormonal evaluation demonstrated dramatically decreased plasma levels of testosterone. COMMENTARY: The clinical, X ray and histologic aspects of this case, although rare, show that the presence of virilization should lead to a search for an androgen-secreting ovarian or adrenal tumor. PMID- 11908172 TI - [Tumor of the nose]. PMID- 11908173 TI - [Serpiginous papular eruption]. PMID- 11908174 TI - [Epidemiology and statistics. Misleading figure]. PMID- 11908175 TI - [Contact hypersensitivity to colophony]. PMID- 11908176 TI - [Human papillomavirus and skin carcinoma in organ transplants. Recent studies]. PMID- 11908177 TI - [Paraneoplastic pemphigus]. PMID- 11908178 TI - [Nursing protocol of chronic wounds for nurses. General principles and techniques]. PMID- 11908179 TI - [Gout tophus]. PMID- 11908180 TI - [Anthrax]. PMID- 11908181 TI - [Post-corticoid atrophy]. PMID- 11908182 TI - [Cardiac liver and itching]. PMID- 11908183 TI - [Treatment with acitretin and alcohol drinking: a high risk combination]. PMID- 11908184 TI - [Edouard Jeanselme (1858-1935)]. PMID- 11908185 TI - [Can treatment of flat angiomas be proposed in the first months of life?]. PMID- 11908186 TI - [Corticosteroids for treating Kawasaki disease]. PMID- 11908187 TI - Just do the right thing. PMID- 11908188 TI - Cerebral amyloid angiopathy as a cause of recurrent intracerebral hemorrhage. AB - Cerebral amyloid angiopathy is a largely untreatable disease often not diagnosed until autopsy. Severe cerebral amyloid angiopathy can cause lobar cerebral hemorrhage, transient neurological symptoms, and dementia with leukoencephalopathy. Several outcome studies have suggested lower mortality and better functional recovery in lobar compared with deep hemorrhage. Recurrence of lobar hemorrhage however is relatively common. We describe a case of recurrent spontaneous intracerebral hemorrhage in a 72-year-old woman requiring surgical evacuations. A review of the literature summarizing the diagnosis and treatment of cerebral amyloid angiopathy is presented. PMID- 11908189 TI - Cervical pregnancy: two case reports. AB - REASON FOR STUDY: To determine if there is a safe and effective treatment for cervical pregnancy. MAIN FINDINGS: There is no study presently which shows whether the medical vs surgical vs medical and surgical treatment combined for cervical pregnancy is more efficient. Continued studies are necessary to validate any treatment and confirm which would increase survival with fewer complications. PRINCIPAL CONCLUSION: There is no definitive treatment for cervical ectopic pregnancies. The risk and benefits of each treatment must be entertained before a decision is made on either medical or surgical treatment. PMID- 11908190 TI - Mentally handicapped institutionalized patients: a five-year review of gynecologic examinations performed under general anesthesia. AB - The validity of the practice of routine examination of mentally handicapped women under general anesthesia was evaluated via a retrospective chart review at Yale New Haven hospital for a five-year period. In the absence of a history of abnormal bleeding or discharge, this select population of low risk women had a zero incidence of gonorrhea and chlamydia, and no cervical dysplasia. PMID- 11908191 TI - Health Insurance Portability and Accountability Act of 1996 (HIPAA): a provider's overview of new privacy regulations. AB - When it enacted The Health Insurance Portability and Accountability Act of 1996, Congress mandated establishment of privacy regulations covering individual health information. Title II of HIPAA, the Privacy Rule that became effective on April 14, 2001, offers Americans the first-ever set of comprehensive protections against the unintended and/or inappropriate disclosure of personal health information. Provisions of the Privacy Rule and its associated regulations include patient control over the use of health information, patient rights to information on the disclosure policies of the health-care provider, patient rights to review and amend one's medical information, standards for limiting the scope of data disclosed to other health-care providers, and penalties for noncompliance with the law. This paper presents a summary of the need for protection of personal health information and an overview of the provisions of this legislative foundation for protecting personal health records--the HIPAA Privacy Rule. PMID- 11908192 TI - Basic principles in the treatment of degenerative spinal disease. PMID- 11908193 TI - Development of allogeneic hematopoietic stem cell transplantation (HSCT). AB - The field of BMT has entered a new phase based on insights into basic molecular and cellular biology. The mechanism of cure mediated by allogeneic HSCT is now believed to be at least partially mediated by cellular attack against tumor associated antigens. Better understanding of these cellular targets and the means of targeting them specifically will allow a more precise application of cellular therapy than is currently possible. This in turn may make it possible to design therapies that are increasingly selective without the collateral toxicities of GVHD, infection, and direct organ damage that currently are the limiting features of allogeneic HSCT as practiced until recently. However, it remains a challenge to demonstrate that disease-control using reduced intensity preparative regimens is at least comparable to that achieved by traditional myeloablative HSCT. This will require carefully controlled studies in each the major diseases that have shown sensitivity to the immune-mediated effects of HSCT. The chapters below will highlight progress towards the goals of elucidating the role of NST. PMID- 11908194 TI - Combined use of autografting and non-myeloablative allografting for the treatment of hematologic malignancies and metastatic breast cancer. PMID- 11908196 TI - Non-myeloablative allogeneic hematopoietic transplantation and induction of graft versus-malignancy. PMID- 11908195 TI - Non-myeloablative hematopoietic stem cell transplantation (NST) in the treatment of human malignancies: from animal models to clinical practice. AB - NST is becoming a widely accepted method for allogeneic HSCT. Much experience has been gained, and the biology, indications and limitations are becoming clearer. Nonmyeloablative conditioning allows consistent engraftment of allografts from matched related, unrelated, and even partially matched donors. NST has been able to reduce the toxicity of allogeneic HSCT. The better immediate outcome produces better overall DFS. NST was feasible in elderly patients with almost no upper age limit, and in patients with organ dysfunction or other comorbidities precluding standard ablative conditioning. NST has also reduced the regimen-related toxicity of allogeneic HSCT in high-risk setting such as HSCT in heavily pretreated patients or following failure of a prior transplant procedure and in the unrelated setting. NST is rapidly becoming the treatment of choice in these indications where toxicity of standard ablative therapy is unacceptable. In certain malignancies such as in NHL, Hodgkin's disease and multiple myeloma, standard ablative NST has been reported to result in exceptionally high treatment related mortality, and NST is being investigated as a more reasonable alternative. NST may reduce the toxicity of the procedure even in younger patients who are eligible for ablative HSCT as well, however the long-term impact on patient outcome in this group is not yet established, and NST merits further investigation in prospective comparative trials. As described above, the known susceptibility of the underlying malignancy to GVT, the response to prior chemotherapy and bulk of residual disease, and the type of donor are other factors to consider when considering NST, and when selecting a regimen. The optimal preparative regimen needs to be defined. Ultimately less chemotherapy will be used and more specific immune-modulation, rather than intense nonspecific immunosuppression, will be used to achieve HVG tolerance. Preliminary animal models using costimulation blockade for specific induction of tolerance are promising steps towards achievement of this goal. Although much progress has been achieved with consistent achievement of engraftment with NST, GVHD and disease recurrence remain major obstacles to successful treatment. Existing clinical data suggest that NST does limit the incidence and severity of GVHD. Limitation of regimen-related toxicity, and bilateral transplantation tolerance afforded by mixed chimerism, are believed to have a major role in limiting GVHD. However GVHD remains the primary cause of treatment-related mortality. The development of techniques to separate GVHD and GVL are essential for further improvement of NST outcome. Better understanding of the biology and targets of GVHD and GVL may allow the elimination of alloreactive T-cells responsible for GVHD from the graft while retaining T-cells with GVL and infection control potential. Recurrence of the underlying malignancy is a major complication when NST is attempted in patients with chemo-refractory diseases and with high tumor bulk. Reduced toxicity regimens such as the FB/ATG regimen have been somewhat more successful in controlling disease progression until a potent GVT effect is established. However novel approaches are urgently required. NST serves as a platform for cellular immunotherapy. Judicious use of pre-emptive DLI needs to be explored. DLI may be amplified by activation of donor lymphocytes with IL-2 or in vivo administration of IL-2. Identification of tumor antigens will lead the way to ex vivo generation and expansion of tumor specific cytotoxic T-lymphocytes to be used as potent immunotherapy without the hazards of GVHD. Allogeneic transplantation is rapidly changing from administration of supralethal doses of chemotherapy and radiation, trying to physically eliminate the 'last tumor cell', to the more subtle and tolerated sophisticated immunotherapy. This effort will focus on specific induction of HVG tolerance followed by induction of tumor specific GVT effect to cure the underlying malignancy. PMID- 11908197 TI - Outpatient allografting in hematologic malignancies and nonmalignant disorders- applying lessons learned in the canine model to humans. AB - The novel allogeneic transplant approach outlined in this chapter proposes the dual concepts that currently used intensive cytoreductive conditioning programs can be successfully replaced by nonmyeloblative immunosuppression and that stem cell grafts create marrow space for engraftment through subclinical GVH reactions. Immunosuppression can be conceptually divided into two parts, one targeted exclusively towards host cells before transplantation, and the other aimed at both host and donor T lymphocytes after transplantation. The resultant effect is to either establish mutual graft-host tolerance as reflected by stable mixed donor-host chimerism or accomplish complete donor chimerism. Preclinical studies have demonstrated the feasibility of this new approach in the context of MHC-identical transplants in young canines. Early data from the dog model of severe hereditary hemolytic anemia suggest that mixed chimerism can partially correct the phenotypic expression of disease, but that complete chimerism will be necessary to halt the continued hemolytic process of the host type minority red blood cell population. In contrast, mixed chimerism would be expected to correct disease manifestations of other severe hereditary red blood cell disorders such as sickle cell disease. The early results in human patients with hematological malignancies have demonstrated the feasibility of establishing hematopoietic engraftment using a nonmyeloblative conditioning regimen. Using this regimen, transplants were performed in the outpatient setting, and the need for transfusion support was minimal. Preliminary information suggests that mixed chimerism does not appear to be stable in this older patient population, with most patients progressing to full donor chimerism and a small minority rejecting. Complete disease responses and molecular remissions have been observed in a significant proportion of patients, suggesting that adoptive immunotherapy may not be necessary in most cases. However, in those patients that engraft with at least mixed chimerism and develop progression of the underlying malignancy, DLIs can be used for subsequent adoptive immunotherapy. The feasibility and safety of the outpatient approach for MHC-nonidentical transplantation has been demonstrated. Graft rejection appears to have been corrected by the addition of fludarabine to the nonmyeloablative conditioning regimen for the recipients of HLA-identical sibling allografts. Despite most recipients of MHC-nonidentical transplantation having sustained engraftment, some rejections have been observed. Studies are ongoing to identify the risk factors for rejection after unrelated HSCT to ensure uniform engraftment in future studies. PMID- 11908198 TI - Non-myeloablative transplants for congenital diseases. AB - The morbidity and mortality associated with postnatal HSCT, toxicity of HSCT conditioning regimens, lifelong immunosuppressive therapy, and lack of compatible donors discourages many patients and physicians from utilizing postnatal HSCT as a treatment for congenital disease. Non-myeloblative in utero HSCT is now being considered as an alternate treatment with the hope that it will be more therapeutic with less toxicity to a wider spectrum of patients with congenital disorders. Prenatal stem cell transfer may eliminate many of the risks and hazards associated with postnatal HSCT, as the fetus may be less reactive than an immunologically mature individual such that tolerance to donor cells could be developed. GVHD and rejection of postnatal therapeutic grafts may be minimized thus reducing or eliminating altogether the need for postnatal myeloablation and immunosuppression. Much work must be done both in animal studies as well as in clinical trials. By using well-designed murine models such as the beta thalassemic mouse outlined above, we believe we can determine the optimal conditions for non-myeloablative postnatal transplants with allogeneic or haplocompatible HSC following prenatal tolerance induction with these cells. In addition, we may answer basic immunology questions regarding the development and regulation of immunity and tolerance in both mice and humans. PMID- 11908199 TI - Immunosuppression with limited toxicity: the characteristics of nucleoside analogs and anti-lymphocyte antibodies used in non-myeloablative hematopoietic cell transplantation. AB - The availability of two groups of pharmacologic agents, the nucleoside analogs and anti-lymphocyte antibody preparations of various specificities has enabled the development of NST. Although these agents are significantly less cytotoxic than high-dose alkylating agents and total-body irradiation (TBI), they are profoundly immunosuppressive. Opportunistic infections such as the reactivation of cytomegalovirus remain clinical obstacles when NST are performed using these agents, especially in elderly and previously immunosuppressed patients. For anti lymphocyte antibody preparations, the degree of immunosuppression produced and hence the risk of opportunistic infection varies depending upon specificity of the antibody (broad versus narrow). Allergic reactions and infusion related toxicity can occur with any antibody preparation, but the infusion of muromonab CD3 is sometimes associated with a particularly potent cytokine-release syndrome. Although fludarabine has been the mainstay of nucleoside analog usage for NST, the other nucleoside analogs-cladribine and pentostatin are beginning to be investigated in this context. PMID- 11908200 TI - Mobilization of allogeneic peripheral blood progenitor cells. AB - It is possible to reliably obtain sufficient PBSC from most normal donors to perform allogeneic transplantation. The mobilization regimen, usually administration of a single daily dose of G-CSF at 7.5 to 10 micrograms/kg subcutaneously for 4 to 6 days, is tolerable with acceptable side effects. However, there is wide variability among individuals with respect to the extent of mobilization achieved by the regimen and the optimal timing of apheresis. Studies suggest that the likelihood of obtaining an adequate harvest of CD34+ cells, as defined locally may be enhanced by employing higher doses or different schedules of G-CSF, monitoring the mobilization and/or collection of PBPC, and using apheresis procedures processing 2 or more times blood volume. However, an optimal regimen for mobilization and harvesting for all donors has not yet been identified and a small percentage of donors may not mobilize adequately with G CSF. Alternative regimens employing combinations of G-CSF and GM-CSF are available that may prove useful in such cases and novel cytokines that are even more effective than G-CSF in mobilizing stem cells are eagerly awaited. Based on currently available experience with normal donors, the short-term safety of G-CSF appears to be acceptable, however there exist several scenarios in which marrow harvesting may be preferable to G-CSF mobilization and apheresis collection of PBPC. PMID- 11908202 TI - Volunteering in Vietnam. PMID- 11908201 TI - Non-myeloblative induction of mixed hematopoietic chimerism: application to transplantation tolerance and hematologic malignancies in experimental and clinical studies. PMID- 11908203 TI - Beyond persevering. PMID- 11908204 TI - What can the Cancer Helpline offer nurses as clinicians? PMID- 11908205 TI - Program addresses surgical nursing needs. PMID- 11908206 TI - More on pensions. PMID- 11908207 TI - Meeting the challenge of distance learning. PMID- 11908209 TI - Creativity in clinical practice. PMID- 11908208 TI - Nursing is pivotal to the nation. PMID- 11908210 TI - Ground-based satellite-type images of the upper-atmosphere emissive layer. AB - With the introduction of infrared (IR) retina sensors used as focal-plane arrays in large telescopes, astronomical observations are now frequently located in the near-IR part of the spectrum. In this region the upper atmosphere introduces in the 0.7-3 microns range an additional component due to the OH vibrational band emission that should be subtracted from the astronomical data. Observations of this upper-atmosphere emission performed at the Pic de Chateaurenard (altitude of 2989 m) are presented here. A panoramic image of the emission is constructed by use of a set of 48 images obtained with a CCD camera mounted on an alt-azimuthal platform. After a numerical filter is used to suppress the star images, the atmospheric emission shows two distinct sets of arches vanishing at two opposite points in the WNW and ESE azimuths. The emissive layer, caused by the ozone hydrogen reaction, is thin and located at the altitude of 85 km. By use of these data, the perspective effect that produces the panoramic arches is inverted in introducing the concept of a virtual camera. The Van Rhijn effect and the refraction correction are taken into account. The three punctual transformations that use matrix algorithms are analyzed. The result is a satellite-type view of the emissive layer that appears as a disk having a radius of approximately 1100 km. This disk is limited by the summit line of the Alps surrounding the Pic de Chateaurenard. The field of view covers a large part of Europe, the Mediterranean Sea, and North Africa. It shows an extended wave system. The images presented show that the upper-atmospheric layer is an efficient tracer of the dynamic processes at that level. Satellite-type views can be calculated without the drawback of looking downward from a satellite and measuring the numerous emissions from cities, oil fields, and other luminous sources. PMID- 11908211 TI - All-optical code-division-multiplexing technique supporting multirate data communications and local-area-network interconnections. AB - An efficient all-optical code-division-multiplexing (AOCDM) technique is proposed to support multirate data communications and local-area-network (LAN) interconnections with multiple protocols. To achieve this goal, we use a strict optical orthogonal code (OOC) in multirate AOCDM systems to guarantee that both cross- and autocorrelation constraints are minimum (i.e., 1) for incoherent optical processing. In contrast, the use of a conventional OOC may result in correlation constraints of 2, which in turn can degrade system performance. Moreover, implementation issues on AOCDM systems are discussed. These include the design of low-cost optical transmitters and the trade-off among coherence time, spectral width, and pulse width for AOCDM systems. It is shown that multirate AOCDM systems have a high operation flexibility to support data communications and LAN interconnections of both equal and multiple bit rates. For multirate data transmissions, the proposed system can have a better bandwidth efficiency and a lower bit error rate than a system that uses a conventional OOC. PMID- 11908212 TI - Comparison of holographic photopolymer materials by use of analytic nonlocal diffusion models. AB - The one-dimensional diffusion equation governing holographic grating formation in photopolymers, which includes both nonlocal material response and generalized dependence of the rate of polymerization on the illuminating intensity, has been previously solved under the two-harmonic expansion assumption. The resulting analytic expressions for the monomer and polymer concentrations have been derived and their ranges of validity tested in comparison with the more accurate numerical four-harmonic case. We used these analytic expressions to carry out a study of experimental results presented in the literature over a 30-year period. Automatic fitting of the data with these formulas allows material parameters, including the nonlocal chain-length variance sigma, to be estimated. In this way, (i) a quantitative comparison of different materials can be made, and (ii) a standard form of experimental result presentation is proposed to facilitate such a procedure. PMID- 11908213 TI - Computational reconstruction of images from holograms. AB - The equations to reconstruct an image plane from a hologram are developed. This development is carried out for planes parallel to the hologram, which allows fast computation through the use of fast Fourier transforms. Algorithms for a digital computer are developed so images can be reconstructed, both with and without the Fresnel approximation, from a digitized hologram without the need for three dimensional optical reconstruction equipment. Examples of holographically recorded images of marine micro-organisms are shown. A computational method for counting the number of micro-organisms in the holographically recorded volume is developed, and an example is provided. PMID- 11908214 TI - Digital watermarking by a holographic technique. AB - A holographic technique is applied for digital watermarking by a computer. A digital-watermark image to be hidden is phase modulated in a random fashion, and its Fourier-transformed hologram is superposed on a content image. The watermark is reconstructed by means of a holographic-reconstruction technique from the bit map image that hides it. In the study the processes of constructing and reconstructing a digital hologram are described on the basis of the theory of Fourier optics. The conditions for superposing the hologram onto the content images are investigated in detail. The validity of the present method is verified by changing the weighting of the hologram relative to that of the content image. The effect of image size is also discussed with respect to reconstruction of the watermark, and it is shown that watermark information in a form of a diffuse-type Fourier-transform hologram cannot be removed by cutting it out of the host image. PMID- 11908215 TI - Target segmentation in active polarimetric images by use of statistical active contours. AB - We address the problem of target segmentation in active polarimetric images, which can reveal contrasts that do not appear in standard intensity images. However, these images are perturbed by strong specklelike noise. For the purpose of segmentation we thus use statistical active contours, which are known to possess noise robustness properties. The polarimetric imagers we consider acquire two different images of the same scene so as to form a two-channel image (TCI). These two images can be combined to form the orthogonal state contrast image (OSCI), which represents the degree of polarization of the backscattered light if its coherency matrix is diagonal. We characterize the segmentation performance of the statistical active contour procedure on the TCI and on the OSCI. In particular, we show that if the illumination beam is spatially nonuniform, it is more efficient to perform the segmentation on the OSCI, which is independent of the spatial variations of the illumination. PMID- 11908216 TI - Magneto-optical disk drive technology using multiple fiber-coupled flying optical heads. Part II. Laser noise considerations. AB - A magneto-optical data storage system utilizing single-mode fiber is capable of providing high signal-to-noise ratio (SNR) recording if laser noise sources are properly managed. In particular, mode partition noise (MPN) associated with use of a Fabry-Perot laser diode can be a significant problem in a fiber-based system. The various mechanisms leading to MPN as well as to laser phase noise are discussed in the context of a system constructed with polarization-maintaining fiber. The primary noise mechanisms include spurious fiber-endface reflections and errors in the quarter-wave plate on the recording head. An understanding of these effects is essential for fabrication of a fiber-based recording system with suitable SNR performance. PMID- 11908217 TI - Model of linewidth for laser writing on a photoresist. AB - We present a theoretical model to describe the feature size produced by direct laser writing upon a photoresist relative to various experimental parameters. The model allows the number of parameters required for describing the linewidth to be reduced and shows how the description can be made in terms of the ratio of laser power to writing velocity. Both of the limiting cases of the truncation of the laser beam are analyzed; i.e., the case of a nontruncated (Gaussian) beam and the case of a strongly truncated beam (simplified with uniform illumination assumed). Experimental measurements are presented that are fitted to the model to permit its validity to be assessed and for a comparison of these two regimes, which are shown to be different. PMID- 11908218 TI - Transfer of micro-optical structures into GaAs by use of inductively coupled plasma dry etching. AB - The transfer of continuous-relief micro-optical structures from resist into GaAs by the use of direct-write electron-beam (e-beam) lithography followed by dry etching in an inductively coupled plasma is demonstrated. BCl3-Ar chemistry was found to give satisfactory results; N2 and Cl2 were added to change the selectivity between GaAs and e-beam resist. The transfer process generates smooth etched structures. Distortion of the diffractive structures in the transfer process was examined. Blazed gratings with a period of 10 microns were optically evaluated with a 940-nm VCSEL. This grating was a five-step approximation of a blazed profile. The diffraction efficiency was 67% in the first order, with a theoretical value of 87%. Also, simulations of the optical performance of the transferred diffractive elements were made by use of a Fourier transform of the grating profile. Our goal is to integrate micro-optical structures with VCSELs. PMID- 11908219 TI - Simultaneous retrieval of atmospheric profiles, land-surface temperature, and surface emissivity from Moderate-Resolution Imaging Spectroradiometer thermal infrared data: extension of a two-step physical algorithm. AB - An extension to the two-step physical retrieval algorithm was developed. Combined clear-sky multitemporal and multispectral observations were used to retrieve the atmospheric temperature-humidity profile, land-surface temperature, and surface emissivities in the midwave (3-5 microns) and long-wave (8-14.5 microns) regions. The extended algorithm was tested with both simulated and real data from the Moderate-Resolution Imaging Spectroradiometer (MODIS) Airborne Simulator. A sensitivity study and error analysis demonstrate that retrieval performance is improved by the extended algorithm. The extended algorithm is relatively insensitive to the uncertainties simulated for the real observations. The extended algorithm was also applied to real MODIS daytime and nighttime observations and showed that it is capable of retrieving medium-scale atmospheric temperature water vapor and retrieving surface temperature emissivity with retrieval accuracy similar to that achieved by the Geostationary Operational Environmental Satellite (GOES) but at a spatial resolution higher than that of GOES. PMID- 11908220 TI - Mechanisms of disease pathogenesis in amyotrophic lateral sclerosis. A central role for calcium. PMID- 11908221 TI - Immune-mediated neuropathies. PMID- 11908222 TI - Molecular basis of hereditary neuropathies. PMID- 11908224 TI - Myasthenia gravis. PMID- 11908223 TI - Neuropathic pain. PMID- 11908225 TI - The Lambert-Eaton myasthenic syndrome. PMID- 11908226 TI - Congenital myasthenic syndromes. PMID- 11908227 TI - Genetic aspects of amyotrophic lateral sclerosis. PMID- 11908228 TI - Metabolic myopathies. PMID- 11908229 TI - Periodic paralyses and nondystrophic myotonias. PMID- 11908230 TI - Inflammatory myopathies. Recent advances in pathogenesis and therapy. PMID- 11908231 TI - Limb-girdle muscular dystrophies. PMID- 11908233 TI - Neuromuscular syndromes associated with human immunodeficiency virus infection. PMID- 11908232 TI - Myotonic dystrophies. PMID- 11908234 TI - Critical illness neuromyopathies. PMID- 11908235 TI - Motor unit number estimation in neurologic disease. AB - Since its introduction 30 years ago, MUNE technologies have been increasingly refined and applied to a wide variety of neuromuscular disorders. Differences of opinion remain among MUNE investigators as to which method should be used; however, statistical and MPS MUNE currently enjoy the most widespread use. A number of methodological issues remain, including the development of detailed universal standards for each technique and modifications for the further enhancement of reproducibility. These issues are the subject of ongoing investigation. However, despite technical variability, the MUNE values obtained with different methods show good agreement, both in studies of healthy subjects and in patients with a variety of neurogenic processes. MUNE has been most successfully applied to patients with ALS and in animal models of motor neuron disease, providing significant insight into the pathophysiology of these disorders. These techniques are being increasingly incorporated into clinical therapeutic trials. MUNE is a technology offering important promise in the study of neuromuscular disease, enabling the collection of novel data in the living patient unobtainable by any other method. PMID- 11908236 TI - Diagnostic criteria and outcome measurement of amyotrophic lateral sclerosis. AB - ALS is a most challenging disease. Accurate diagnostic criteria are well along in refinement, and experience with patients who are at the limits of the criteria should be presented to increase our comfort with the limits of ALS or motor neuron disease. As our understanding of the pathophysiology is further refined, there will be new drugs to test. It will be important to have sensitive clinical and electrophysiologic measures available to detect changes in the rate of progression. When effective drugs are identified, they will, in addition to helping patients, lend support to proposed mechanisms. Charcot would be impressed with our advances in the ability to detect and follow upper and lower motor neuron loss, but would urge us to press on in our clinical endeavors. PMID- 11908237 TI - Drug therapy in amyotrophic lateral sclerosis. PMID- 11908238 TI - Spinal muscular atrophies. PMID- 11908239 TI - Diabetic neuropathy. PMID- 11908240 TI - Legislation calls for Medicare coverage of outpatient drugs, pharmacists' services. PMID- 11908241 TI - Black-box warning for droperidol surprises pharmacists. PMID- 11908242 TI - Bosentan enters market with risk management program. PMID- 11908243 TI - Most clinical practice guideline authors receive drug industry support. PMID- 11908244 TI - Value on the margin? PMID- 11908245 TI - Prevalence and cost savings of therapeutic interchange among U.S. hospitals. AB - The prevalence and cost savings of therapeutic interchange (TI) among teaching, nonteaching, and investor-owned hospitals in the United States was studied. A survey was sent to all directors of pharmacy at hospitals listed in the 1999 American Hospital Association directory as having more than 100 beds; 463 (29.8%) hospitals responded. The survey elicited data about hospital demographics, the policies and personnel involved in TI, and the estimated cost savings incurred by the use of TI. Eighty-eight percent of teaching, 89% of nonteaching, and 100% of investor-owned hospitals reported having established TI policies and procedures; 88% of responding hospitals reported the use of TI as a means of formulary management. Individuals involved in the decision-making process for TI policies included physicians, pharmacists, and pharmacy and therapeutics committee members. Most responding hospitals reported having an automatic interchange procedure, and few required physician consent before the substitution was made. The most commonly substituted medication classes were histamine H2-receptor antagonists, proton-pump inhibitors, antacids, and quinolones. Differences in TI procedures and the medication classes commonly substituted were not significant between teaching and nonteaching hospitals. The annual dollar savings was estimated by 36% of teaching, 38% of nonteaching, and 50% of investor-owned hospitals and determined by record keeping in 18% of teaching, 20% of nonteaching, and 40% of investor-owned hospitals. Eighty-eight percent of teaching, 89% of nonteaching, and 100% of investor-owned hospitals have established TI policies. Significant variation in cost savings occurred when hospitals attempted to estimate the annual dollar savings, as no adequate commercially available software exists to perform this task. PMID- 11908246 TI - Survey of hospital policies regarding low-molecular-weight heparins. AB - Hospital policies regarding the use of low-molecular-weight heparins (LMWHs) were studied. A questionnaire addressing the formulary status of LMWH products, the use of prescribing guidelines, programs for therapeutic interchange, and policies to promote alternatives to LMWHs when appropriate was prepared. The questionnaire was mailed in January 2001 to pharmacy directors at 70 hospitals located in 19 states. All the hospitals were members of a national group purchasing organization. Forty-nine usable responses were received, for a response rate of 70%. Enoxaparin and dalteparin were the LMWH products most likely to be on the respondents' formularies (98% and 29% of hospitals, respectively). About 29% of the hospitals reported having guidelines on the use of LMWHs. Among hospitals that did not, most indicated that they were considering or would like to implement such guidelines. The most commonly cited barrier to the development and implementation of guidelines was lack of pharmacy personnel. Ten percent of the respondents reported having therapeutic-interchange programs for LMWHs. Cited barriers to therapeutic interchange programs included lack of therapeutic equivalence among products and lack of comparable labeled indications. Policies to promote alternatives to LMWHs were reported by 18% of the respondents. A multihospital survey showed that many hospitals wanted but relatively few had prescribing guidelines for LMWHs. PMID- 11908247 TI - Stability of irinotecan hydrochloride in aqueous solutions. AB - The stability of irinotecan after reconstitution in several vehicles for i.v. infusion was studied. Irinotecan hydrochloride injection was diluted in phosphate buffer solution (pH 4.0, 6.0, and 7.4), 5% dextrose injection, and 0.9% sodium chloride injection to a final concentration of 20 micrograms/mL. The solutions were stored at 25, 37, and 50 degrees C and assayed at intervals up to 24 hours by high-performance liquid chromatography for the concentration of the lactone form of irinotecan remaining. The effect of temperature and pH on the extent and rate of degradation of irinotecan was determined. The hydrolysis of irinotecan to its carboxylate form was reversible. The rate and extent of hydrolysis increased with increasing pH. The use of a weakly acidic vehicle, such as 5% dextrose injection, for reconstitution of irinotecan may maintain the drug's stability prior to administration. PMID- 11908248 TI - Determining hyperforin and hypericin content in eight brands of St. John's wort. PMID- 11908249 TI - Lipid-lowering therapy at hospital discharge after coronary artery bypass grafting. PMID- 11908250 TI - Internal approach to competency-based credentialing for hospital clinical pharmacists. PMID- 11908251 TI - Profession of second lieutenants. PMID- 11908252 TI - From the philosophy auditorium to the neurophysiology laboratory and back: from Bergson to Damasio. AB - Henri Bergson (1859-1941) was probably the most influential French philosopher at the turn of the twentieth century. In 1927 he was awarded the Nobel Prize for literature. Far beyond the restricted academic philosophical milieu, the impact of his thinking reached personalities as diverse as Claude Debussy, Marcel Proust, George Bemard Shaw, and the impressionists. His essay The Laughter (Le Rire) is one of the most profound and original ever written on the sense of humor. Bergson's opinions, with their emphasis on life, instinct and intuition, represented a deviation from the rationalist mainstream of western philosophical tradition. In some circles he was received with skepticism and irony, as in Bertrand Russel's History of Western Philosophy. Today, unbiased by theoretical "bergsonism," neurophysiologic research--as undertaken mainly by Antonio Damasio's team at Iowa University--confirms many of his hypotheses and elucidates their mechanisms. In this new light, intuition and "recognition by the body" should not be seen as the personal fantasy of an original thinker but as fundamental cognitive tools. PMID- 11908253 TI - Novel insights into the natural history of apical hypertrophic cardiomyopathy during long-term follow-up. AB - BACKGROUND: Long-term follow-up in apical hypertrophic cardiomyopathy is rare. OBJECTIVE: To study the natural history of the disease. METHODS: We followed 11 patients, 5 women and 6 men, for 5-20 years. RESULTS: At presentation all 11 patients had typical features of apical hypertrophic cardiomyopathy, with dyspnea in 3 and chest pains in 8, of whom 5 were typical of angina and 3 had myocardial infarction. R-wave voltage and T-wave negativity progressively decreased in magnitude at serial electrocardiograms in four patients. Perfusion defects were detected on thallium myocardial scintigraphy in three, increased apical uptake in two, and normal in one patient. Apical aneurysm with normal coronary arteries developed in a patient who had sustained ventricular tachycardia. All of the 10 catheterized patients had normal coronaries except for one with significant left anterior descending artery stenosis and another with a minor lesion. Symptomatic sustained ventricular tachycardia was found in two patients, one of whom required the implantation of an internal cardioverter defibrillator. CONCLUSIONS: Apical hypertrophic cardiomyopathy may develop morphologic and electrocardiographic changes with life-threatening arrhythmias necessitating close follow-up and treatment. PMID- 11908254 TI - Management by constraints: considering patient volume when adding medical staff to the emergency department. AB - BACKGROUND: The emergency department is one of the hospital's busiest facilities and is frequently described as a bottleneck. Management by constraint is a managerial methodology that helps to focus on the most critical issues by identifying such bottlenecks. Based on this theory, the benefit of adding medical staff may depend on whether or not physician availability is the bottleneck in the system. OBJECTIVE: To formulate a dynamic statistical model to forecast the need for allocating additional medical staff to improve the efficacy of work in the emergency department, taking into account patient volume. METHODS: The daily number of non-trauma admissions to the general ED was assessed for the period 1 January 1992 to 1 December 1995 using the hospital computerized database. The marginal benefit to shortening patient length of stay in the ED by adding a physician during the evening shift was examined for different patient volumes. Data were analyzed with the SAS software package using a Gross Linear Model. RESULTS: The addition of a physician to the ED staff from noon to midnight significantly shortened patient LOS: an average decrease of 6.61 minutes for 80 119 admissions (P < 0.001). However, for less than 80 or more than 120 admissions, adding a physician did not have a significant effect on LOS in the ED. CONCLUSIONS: The dynamic model formulated in this study shows that patient volume determines the effectiveness of investing manpower in the ED. Identifying bottleneck critical factors, as suggested by the theory of constraints, may be useful for planning and coordinating emergency services that operate under stressful and unpredictable conditions. Consideration of patient volume may also provide ED managers with a logical basis for staffing and resource allocation. PMID- 11908255 TI - Bladder tumor antigen stat test in non-urothelial malignant urologic conditions. AB - BACKGROUND: The bladder tumor antigen stat is a simple and fast one-step immunochromatographic assay for the detection of bladder tumor-associated antigen in urine. OBJECTIVES: To evaluate the BTA stat in non-bladder cancer patients in order to identify the categories contributing to its low specificity. METHODS: A single voided urine sample was collected from 45 patients treated in the urology clinic for conditions not related to bladder cancer. Each urine sample was examined by the BTA stat test and cytology. RESULTS: The overall specificity of the BTA stat test was 44%, which was significantly lower than that of urine cytology, 90%. The false positive rates for the BTA stat test varied among the different clinical categories, being highest in cases of urinary tract calculi (90%), and benign prostatic hypertrophy (73%). Exclusion of these categories from data analysis improved BTA stat specificity to 66%. CONCLUSIONS: Clinical categories contributing to low BTA stat specificity can be identified, and their exclusion improves the specificity of this test. PMID- 11908256 TI - Experience of hormonal therapy with anastrozole for previously treated metastatic breast cancer. AB - BACKGROUND: Recent years have brought significant progress to the development of hormonal therapies for the treatment of breast cancer. Several new agents have been approved for the treatment of breast cancer in the metastatic setting, among which is the new non-steroidal aromatase inhibitor, anastrozole, introduced for clinical use in Israel in March 1997. OBJECTIVE: To evaluate the response rate and survival duration of patients treated with anastrozole for metastatic breast cancer, who had previously received at least one line of hormonal therapy. METHODS: Anastrozole was administered to 37 patients with metastatic breast cancer. The median age was 64 years. Estrogen receptor was positive in 20 patients, negative in 10 and unknown in 7. All patients were previously treated with tamoxifen in the adjuvant setting or as first-line hormonal therapy for metastatic disease. Anastrozole was given orally, 1 mg/day. Response was evaluated 2 months after the initiation of treatment and reevaluated every 2 months. Therapy was given until disease progression. Ten ER-negative patients were excluded from the final analysis. RESULTS: Twenty-seven patients were eligible for response and toxicity analysis. The median follow-up was 20 months. One patient (3.7%) achieved complete response and remains free of disease 28 months after start of therapy. No partial responses were seen. Twenty patients (74%) had stable disease. Two year actuarial survival was 57%. Median survival was 26.5 months after starting therapy and median progression-free survival was 11 months. The toxicity was mild: one patient (3.7%) complained of weight gain and one patient (3.7%) had mild fatigue. CONCLUSION: Although the response rate was low, hormonal therapy with anastrozole seems to be beneficial in terms of disease stabilization, freedom from progression, and overall survival without serious toxicity. PMID- 11908257 TI - The effect of meperidine and promethazine on fetal heart rate indices during the active phase of labor. AB - BACKGROUND: Visual interpretation of fetal heart rare monitoring is subject to intra- and inter-observer variability. OBJECTIVE: To examine the effect of intrapartum administration of meperidine and promethazine on fetal heart activity measured objectively by a computerized system. METHODS: Fourteen healthy women with normal pregnancies at term were studied during the active phase of labor. Fetal heart rate was recorded with the Oxford Sonicaid system 8000. Recordings were performed for 40 minutes prior to and after maternal intravenous administration of meperidine 50 mg with promethazine 25 mg. RESULTS: The combination of meperidine and promethazine caused a significant decrease in the number of accelerations of 10 beats per minute (9.7 versus 2.6, P = 0.002) and 15 beats per minute (5.2 vs. 1.4, P = 0.003), time spent in episodes of high variation (14.8 vs. 2.0, P = 0.005) and short-term variation (7.8 vs. 5.0, P = 0.003). On the other hand there was an increase in the time spent in episodes of low variation (5.3 vs. 19.7, P = 0.009). CONCLUSIONS: Maternal administration of meperidine with promethazine has a significant effect on FHR indices during the active phase of normal labor. PMID- 11908258 TI - Pericarditis and pericardial effusion in acute ST-elevation myocardial infarction in the thrombolytic era. AB - BACKGROUND: Myocardial infarction-associated pericarditis is a common cause of chest pain following MI, its frequency depending on how it is defined. OBJECTIVES: To investigate the incidence of acute pericarditis and pericardial effusion in the acute phase of ST-elevation MI treated with thrombolytic therapy. METHODS: The study group comprised 159 consecutive patients fulfilling the criteria for acute MI who were admitted to our department during 18 months. Infarct-associated pericarditis was defined as the finding of a pericardial friction rub, a typical pleuropericardial pain, or both. All patients underwent physical examination of the cardiovascular system four times daily for 7 days, as well as daily electrocardiogram and echo Doppler examinations. RESULTS: Fourteen patients (8.8%) developed a friction rub and 11 patients (6.9%) had a mild pericardial effusion. Six patients (4.0%) had both a friction rub and pericardial effusion. Two patients had a friction rub for more than 7 days. Pleuropericardial chest pain was present in 31 patients (19.5%) but only 7 of them had a friction rub. The in-hospital mortality rate was 1.3% and no mortality was observed in the acute pericarditis group. CONCLUSION: The incidence of signs associated with acute pericarditis was lower in MI patients treated with thrombolysis, compared with historical controls, when a friction rub and/or pericardial effusion was present. There was no significant reduction in the incidence of pleuropericardial chest pain. PMID- 11908259 TI - Nitrofurantoin-induced chronic active hepatitis. AB - BACKGROUND: Nitrofurantoin is a commonly prescribed urinary antiseptic. Hepatic injury has been associated with its use. OBJECTIVES: To present three patients in whom long-term exposure to the drug resulted in chronic active hepatitis; and to review the epidemiology, clinical immunology, histopathology, pathogenetic features and treatment of previously reported cases. RESULTS: Withdrawing nitrofurantoin once the diagnosis was suspected did not lead to remission of the liver disease and glucocorticoids had to be administered. One patient died of liver failure. CONCLUSIONS: Awareness of this unusual side effect of nitrofurantoin is important and caution should be taken before prescribing it. Over the past years new insight into the immune nature of this drug has emerged. PMID- 11908260 TI - Plasma homocysteine: a new risk factor for Alzheimer's disease? PMID- 11908261 TI - Oligonephropathy: from a rare childhood disorder to a possible health problem in the adult. AB - Recent data have shed significant new light on the structural and functional development of the kidneys, as well as on a rare congenital form of bilateral renal hypoplasia called congenital oligomeganephronia. In this renal disorder, few greatly enlarged and "hard-working" nephrons are found that will ultimately sclerose and lead to end-stage renal failure during early childhood. At the same time it has been recognized that the number of nephrons in the kidneys of various animal species and humans is correlated to renal mass. Therefore, premature babies and/or infants small for gestational age due to intrauterine malnutrition will be born with relatively small kidneys and a certain nephron deficit, a condition called congenital oligonephropathy. Extensive worldwide epidemiologic studies have now shown that these premature or SGA infants have a high incidence of cardiovascular disease, hypertension, hyperlipidemia, diabetes and renal failure in adulthood. Although the pathophysiologic mechanisms responsible for these complications of premature birth are not entirely understood, it has become clear that the described association may pose a possible health problem in the adult population. This review describes the background of COMN and CON as well as the evidence that has accumulated on the adult complications of the latter. In addition, some thoughts are presented on the importance of identifying subjects possibly affected by CON, such that early recognition may alter the ultimate outcome. PMID- 11908262 TI - Nitric oxide in asthma. PMID- 11908263 TI - Alcohol consumption and the gastrointestinal tract. AB - Alcohol is one of the most commonly abused drugs, with a per capita consumption of approximately 10 L pure ethanol per year in the United States and even higher in Spain and France. In terms of mortality, the effect of alcohol on the liver and the pancreas is probably more significant than on the tubular gastrointestinal tract. However, alcohol is a very important cause of morbidity in the tubular gastrointestinal tract. Alcohol influences the motility in the esophagus, stomach and small bowel and has direct effects on the mucosa of the upper tract. While the stimulation of gastric acid secretion is inversely correlated with the alcohol concentration of the beverage, a direct pathogenetic role in peptic ulcer disease has not been demonstrated. Some alcohols, like red wine, have been shown to possess an anti-Helicobacter pylori effect. Alcohol also has a role in the development of tumors of the gastrointestinal tract. PMID- 11908264 TI - Apical hypertrophic cardiomyopathy: truly a benign entity or not benignly a true entity? PMID- 11908265 TI - Homocysteine gets to the brain. PMID- 11908266 TI - Leptin and transplantation: pieces are still missing in the puzzle. PMID- 11908267 TI - Mechanical alternatives to the human heart: intracorporeal assist systems and total artificial heart. AB - The currently available intracorporeal assist systems and total artificial heart provide long-term support for patients terminally ill with congestive heart failure, often in an out-of-hospital environment. While they are currently widely used mainly as a long-term bridge to heart transplantation, their inherent risks of infection or stroke are prompting interest in future devices that will be smaller, fully implantable and hopefully stroke-free. These devices will be described in the last part of this review. PMID- 11908268 TI - Serum concentrations of leptin in heart, liver and kidney transplant recipients. AB - BACKGROUND: Leptin is a 16 kDa hormone synthesized by adipocytes and involved in body weight regulation. OBJECTIVES: To determine serum leptin concentrations in heart, liver and kidney transplant recipients. METHODS: We investigated 57 patients: 18 male heart transplant recipients (age 25-69 years) at 1-66 months after transplantation, 6 female and 8 male liver transplant recipients (age 33 70) at 11-73 months after transplantation, and 10 female and 15 male kidney transplant recipients (age 20-61) at 3-138 months after transplantation. All recipients were receiving immunosuppressive therapy, including prednisone 0-20 mg/day, azathioprine 75-125 mg/day, cyclosporin 100-250 mg/day or tacrolimus 2-10 mg/day. The results were compared to those of 10 female and 10 male healthy controls. Morning serum concentrations of leptin were measured with a commercial radioimmunoassay (Linco Research Inc., USA), and serum insulin and cortisol levels were measured by radioimmunoassay. RESULTS: Patients (both men and women) after heart, liver and kidney transplantation exhibited significantly higher serum concentrations of leptin and leptin/body mass index ratios than controls. Serum leptin concentrations were significantly higher in women than in men and correlated very significantly with BMI in all cases. The multivariate stepwise analyses showed that among parameters including BMI, gender, age, time after transplantation, prednisone dose, hematocrit, serum concentrations of glucose, albumin, creatinine, cortisol and insulin, only BMI, gender, cortisol and insulin were significant independent determinants of serum leptin levels in these patients. CONCLUSIONS: This is the first report showing that, in addition to body mass index and gender, basal cortisol and insulin levels affect the hyperleptinemia in transplant patients. The clinical relevance of hyperleptinemia in these patients will require further investigation. PMID- 11908269 TI - Omenn syndrome in the context of other B cell-negative severe combined immunodeficiencies. AB - Severe combined immunodeficiencies represent a heterogeneous group of hereditary defects of the immune system that affect both T and B cells and whose etiology has only recently begun to be understood. A portion of these SCID patients bear a defect in either of the two recombination-activating genes, Rag-1 or Rag-2, while others have mutations in a newly identified gene, Artemis. Omenn syndrome is an unusual severe immunodeficiency with T cells but no B cells, and peculiar features also due to a defect in Rag-1 or Rag-2 genes. All these three forms are characterized by an impairment of the VDJ recombination, the process that insures the somatic diversification of immunoglobulin and T cell receptor-encoding genes. Recent findings have enabled us to better understand the pathophysiology of these three immunodeficiencies, which affect the V(D)J recombination process to a different extent and in different ways. PMID- 11908270 TI - Recurrent stroke in a young patient with celiac disease and hyperhomocysteinemia. PMID- 11908271 TI - Leukemoid reaction associated with transitional cell carcinoma. PMID- 11908272 TI - Acute confusional state developing in a patient taking acyclovir: from the frying pan into the fire. PMID- 11908273 TI - Trichotillomania: a possible therapeutic strategy for the family doctor. PMID- 11908274 TI - Acute hemorrhagic edema of infancy. PMID- 11908275 TI - A huge cystic neck mass in an elderly patient. PMID- 11908276 TI - Acro-osteolysis in a patient with scleroderma. PMID- 11908277 TI - The role of magnesium. PMID- 11908279 TI - Political commitment and public health prioritization. PMID- 11908278 TI - Hospitalization of nursing home residents: the lesson to be studied. PMID- 11908280 TI - Let's be pro-active about this. PMID- 11908282 TI - Opening the way. PMID- 11908281 TI - The journey of a president: Alan Tom DDS. PMID- 11908283 TI - Highlights from the HDS meeting of the memberships. PMID- 11908284 TI - Make the telephone your ally, not your foe! PMID- 11908286 TI - Researchers seek new methods of dental pain management. PMID- 11908285 TI - Three-dimensional computer tomography (CT) in the evaluation of placement for dental implants. PMID- 11908288 TI - Generic dentistry and the sacred trust. PMID- 11908289 TI - HDA endures a tough legislative session. PMID- 11908290 TI - Access to dental care for all our children. PMID- 11908291 TI - A closer look at the proposed national ad campaign. PMID- 11908292 TI - Members awakening to legislative concerns. PMID- 11908293 TI - How the Lt. Governor serves our state, and what we can accomplish together. PMID- 11908294 TI - Do we measure up in service excellence? PMID- 11908295 TI - Clean teeth, healthy heart. PMID- 11908296 TI - Desmoplastic variant of ameloblastoma in an 83-year-ald Asian female: report of a case with literature review. AB - The desmoplastic ameloblastoma is a rare histologic variant of ameloblastoma. This presentation outlines the successful treatment of an 83-year-old Asian female with this type of tumour by surgical resection and peripheral ostectomy with preservation of the interior border of the mandible. Clinical, radiographic, histopathologic and surgical aspects of this tumour were reviewed. PMID- 11908298 TI - Early childhood dental caries in Hawai'i. PMID- 11908297 TI - A comparison of custom-made mouthguards. PMID- 11908299 TI - Early periodic screening diagnosis & treatment. PMID- 11908300 TI - Business demands vs. patient needs: ethical issues challenged by managed care. PMID- 11908301 TI - Esthetic primary anterior crowns. PMID- 11908302 TI - Convention preview: an interview with Ronald D. Jackson, DDS, FAACD. Interview by Dr. Dennis Nagata. PMID- 11908303 TI - Behind in retirement planning? How to make up for lost time. PMID- 11908304 TI - Qualities of a great employee. PMID- 11908305 TI - Establishing and maintaining the "appearance-related" dental practice. PMID- 11908306 TI - Molar-kai express. Flights of fancy of an airborne dentist. PMID- 11908308 TI - The electronic dental office. PMID- 11908307 TI - The doctor in DR. PMID- 11908309 TI - Independent practice associations come to Hawaii. PMID- 11908310 TI - Highlights from the HDS meetings of the memberships. PMID- 11908311 TI - HDA entering the information superhighway. PMID- 11908312 TI - On the fast track.... Electronic claims processing ... putting technology to work for you. PMID- 11908313 TI - Computers becoming necessary component in effective practice management. PMID- 11908314 TI - Treating the elderly patient. PMID- 11908315 TI - Dry mouth (xerostomia). PMID- 11908316 TI - People, PAC's and politics. "It seems to me...". PMID- 11908317 TI - Emergency dental care. PMID- 11908319 TI - Overcoming the enemy. PMID- 11908318 TI - Committee reports. 1995 Hawaii Dental Association Insurance Committee survey results. PMID- 11908320 TI - There is a segment of the Oahu community that is in dire need of dental services. PMID- 11908321 TI - Nursing workforce issues: an international perspective. PMID- 11908322 TI - Addressing workforce issues. PMID- 11908323 TI - Nurses making a difference. PMID- 11908324 TI - Resistance linked to antibiotic misuse. PMID- 11908326 TI - Australian Infection Control Association outlines its activities. PMID- 11908325 TI - Now wash your hands. PMID- 11908327 TI - The challenge of malignant wounds. PMID- 11908328 TI - Nursing in Japan. PMID- 11908329 TI - Time to stop whingeing. PMID- 11908330 TI - Become part of the solution. PMID- 11908331 TI - The nurse's role in the child and family's adherence to asthma treatment. PMID- 11908332 TI - The toothache god of Kathmandu. PMID- 11908333 TI - Committee report. Long Range Planning. PMID- 11908334 TI - [Birth of otology illustrated by original texts]. AB - To be born, to appear, to begin. Though setting the birth of otology in the 19th century may seem a bit pretentious, the texts published at that time are ample proof of the fact. The combined impact of technical, scientific, medical and social factors led to the development of otology as a medical specialty. We discuss these different aspects in this article, illustrating them with extracts from the texts of the era. Once could say that the first half of the 19th century was a period of gestation for otology that was to come into full bloom during the second half of the same century. PMID- 11908335 TI - [Inverted papilloma: endoscopic versus external surgery. Apropos of 28 cases]. AB - To evaluate the role of endoscopic versus external surgery in the treatment of inverted papillomas, the clinical courses of 35 patients over a period of 10 years were reviewed. 13 patients were treated endoscopically whereas 15 were treated with an external approach. 7 patients with a post operative follow up of less than 12 months were excluded from the study. Recurrences occurred in 4 patients, 2 patients had been treated by endoscopic surgery and 2 by medial maxillectomy by lateral rhinotomy. 3 patients were diagnosed with squamous cell carcinoma. Salvage surgery was performed by an external procedure or endoscopically depending on the extension of the recurrence. Late complications occurred in both groups: cosmetic complaints and epiphora were more frequently encountered after external treatment. Functional complaints were noted after endoscopic treatment. If there is no evidence of associated malignancy, if complete exposure of the tumor is possible and long term follow up is feasible, the authors propose endoscopic surgery as first line treatment to excise the body of the tumor, assess it's extension, and remove the root of the tumor. If this is not the case, medial maxillectomy by external approach should be performed. PMID- 11908336 TI - [Cranialization of the frontal sinus]. AB - In the surgery of frontal sinus exclusion, the place of cranialization is still controversial. Our aims were to describe the operative technique, report our results and discuss its indications. PATIENTS AND METHODS: Nineteen patients were operated according to this technique between 1984 and 1997. Cranialization was performed in the first place for tumors, osteitis, traumatisms and benign tumors or mucoceles with a special location. In patients with chronic sinusitis or mucocele, it was performed in the second place when functional surgery was considered as failure and because sinus obliteration seemed to be inappropriate. RESULTS: There was no mortality nor anosmia linked to the procedure. Postoperative sequelae were related to the disease or to the surgical approach. After a median follow-up of 29 months, no disease recurrence was observed. CONCLUSION: Cranialization of frontal sinus gives good results in selected patients, with low morbidity. Even though its indications are infrequent, this technique has its place in the surgical exclusion of frontal sinus. PMID- 11908337 TI - [Is hepatic ultrasonography necessary in the initial check-up of patients with squamous cell carcinoma of the upper respiratory and digestive tract?]. AB - PURPOSE OF THE STUDY: The purpose of our study was to determine the position and value of ultrasound scan of the liver in the initial check-up of patients treated for a squamous cell carcinoma of the upper respiratory and digestive tract. MATERIAL AND METHODS: Our study is based on a retrospective review of 267 patients (249 males and 18 females) managed in the E.N.T. Department of Grenoble universitary hospital from 1993 to 1995 for a upper respiratory and digestive tract malignant tumor. No patient has been previously treated. The site of the primary tumor was: the oropharynx (108 cases), the hypopharynx (88 cases), the oral cavity (44 cases), the larynx (20 cases), the rhinopharynx (6 cases) and the cervical oesophagus (1 case). Endoscopic procedure with biopsy was performed for all the patients. Histologic examination revealed an invasive squamous cell carcinoma in all the cases. The complete check up included a ultrasound scan of the liver and a chest X-ray for all the patients. RESULTS: Ultrasound scan of the liver revealed one or several metastases in 4 cases (1.5%). The primary tumor was hypopharyngeal in 3 cases (2 stages III, 1 stage IV) and oropharyngeal in 1 case (stage III). In three cases, carcinoma was poorly differentiated. Ultrasound scan of the liver was doubtful for 8 patients (3%). The primary tumor was oropharyngeal in 6 cases (1 stage I, 3 stages III, 2 stages IV), laryngeal in 1 case (stage III) and hypopharyngeal in case (stage IV). In six cases carcinoma was well differentiated. All the complementary examinations concluded to a benign liver disease, with a mean diagnosis delay of 4 weeks for the 8 patients. The mean follow-up duration of the 8 patients was 22 months (range 9 to 42 months). None presented any metastases during the follow up. CONCLUSION: Our results compared with those of the literature revealed that ultrasound scan of the liver is a few specific examination which may be recommended for hypopharyngeal tumors, or for a large cervical adenopathy (N2 or N3), a poor differentiated tumor wherever the site of the primary tumor is. PMID- 11908338 TI - [Positron emission tomography in head and neck oncology]. AB - Defining a therapeutic strategy in oncology requires a substantial amount of imaging data provided by modern techniques. While the description of the lesions and their environment has become very precise, there remains a certain degree of uncertainty concerning tissue typing. Positron emission tomography is a scintigraphy technique which can produce quantitative images of metabolic characteristics. 18F-fluorodeoxyglucose is a tracer allowing an analysis of glucose metabolism known to be highly increased in malignant tissues. Increased uptake is an indication of malignancy with an established correlation with proliferative capacity. The only limitation of the method is the generally weak uptake observed in benign hypermetabolic inflammatory or infectious areas. All stages of the disease are concerned for head and neck cancer patients. Clinical experience to date indicates that PET can be applied most usefully to search or residual disease with a possible differentiation between post-therapeutic fibrosis and viable tumor tissue as well as the identification of early relapse. Study of early response to chemotherapy is also an interesting application. Search for extension can also be improved with this technique allowing both regional and whole body explorations. Positron emission tomography is not widely available in France at the present time so all patients cannot be examined with this technique. Positron emission tomography is an evolving technique with improvements being proposed both in the technique and in tracer elements. Further information will be available with new developments in this non-invasive exploration tool. PMID- 11908339 TI - [Extracorporeal lithotripsy in the treatment of salivary lithiasis. A prospective study apropos of 27 cases]. AB - In a prospective study, we evaluated the efficacy of extracorporeal lithotripsy (ECL) in the treatment of salivary stones. We define the indications for the treatment of submaxillary and parotid stones, depending on the diameter and the location of the stone. Twenty-seven patients were treated by ECL. All had a unique stone, in the salivary duct, with a diameter greater than 2 mm; the site was the parotid gland for ten cases, the submaxillary gland for seventeen cases. After six months, the results from a clinical and ultrasonography point of view, were the following: total efficacy or complete desintegration of the stone: 9 cases; partial efficacy or fragmentation of the calculi, with residual fragments with a diameter of less than 2 mm, which could be flushed out with possible spontaneous clearance: 10 cases; failure or the lack of the reduction in the size of the stone or partial fragmentation into concrements with a diameter of more than 2 mm: 8 cases. Based on our experience and the results reported in the medical literature, we propose an therapeutic approach to symptomatic salivary stones. ECL is the most preferable treatment for stones of the parotid duct or parotid gland. The treatment of stones of the submaxillary gland depends on the site and the size of the calculi; ECL should be proposed as first-line treatment when the stone diameter ranges from 2 to 10 mm and when it is situated in the pelvis of the duct or in the proximal duct; surgical resection of the submandibular gland is indicated when the stone is situated in the gland itself or when the diameter is more than 10 mm; when it is situated in the distal duct, a marsupialization should be performed. PMID- 11908340 TI - [Parathyroid cysts]. AB - Two cases of parathyroid cysts are described. One cyst was symptomatic, the other was discovered while performing thyroidectomy. In both cases, the cysts were non functional. There have been reports of functional cysts occurring in a lesser proportion and causing primary hyperthyroidism. These benign cysts are rare (300 cases reported in the literature). Surgery is the treatment of choice in most cases. PMID- 11908341 TI - [Cervical spondylodiscitis: a rare complication of phonatory implants]. AB - Cervical spondylodiscitis is described more often in children than in adults. Many cases occur after cervical or facial surgery, usually as a complication after difficult intubation, endoscopy or foreign body extraction. We report the first case of cervical spondylodiscitis occurring subsequent to secondary tracheoesophageal puncture for implantation of a phonatory prosthesis. Signs were not specific leading to difficult diagnosis and treatment. We discuss the impact of bacterial contamination during tracheoesophageal puncture. PMID- 11908342 TI - [Dysphagia and cervical osteophytosis: case report and review of the literature]. AB - Dysphagia with anterior osteophytosis of the cervical bone is a more and more reported affection, especially after the age of 60. However it remains a rare event. The cause to effect relationship between these two entities should be documented by a complete assessment. Surgical treatment is reserved for patients whose medical treatment has failed. We describe an original case of cervical bone osteophytosis which was diagnosed during an acute and febrile aphagia and review the literature about its assessment and treatment. PMID- 11908343 TI - Current status of nightguard vital bleaching. AB - Indications for using a 10% carbamide peroxide material in a custom-fitted tray to whiten teeth include teeth discolored from aging, chromogenic foods and drinks, and smoking, and also brown fluorosis-stained teeth, single dark teeth, and tetracycline-stained teeth. Tetracycline-stains may require 2 to 6 months of nightly treatment, whereas after discolorations generally resolve in 2 to 6 weeks. After an initial relapse in the first 2 weeks after the end of treatment, color tends to be stable for 1 to 3 years, with some treatments being permanent. The ADA has approved only six 10% carbamide peroxide materials, which have extensive research and publications on safety and efficacy. Considering the average cost of $196 per arch, and the noninvasive nature of this treatment, nightguard vital bleaching is probably the safest, most cost-effective, patient pleasing method to improve the appearance of a smile. However, it should be supervised by a dentist for the proper examination, diagnosis of the cause of discoloration, treatment options, and fabrication and fitting of the carrier. Sensitivity during whitening may be treated with fluoride and potassium nitrate. PMID- 11908344 TI - Nightguard vital bleaching: dark stains and long-term results. AB - Since its introduction to dentistry in 1989, nightguard vital bleaching has proven to be a simple and safe procedure for lightening discolored teeth. Efficacy of the technique is 98% for non-tetracycline-stained teeth, and with extended treatment time, tetracycline-stained teeth can be expected to lighten in 86% of cases. Satisfactory retention of the shade change without additional treatment can be expected in 63% of patients 3 years post-treatment and in at least 42% of patients at 7 years. Side effects are usually mild and transient, disappearing within days of treatment completion. Patients report that they are glad they went through the procedure and 98% recommend the procedure to a friend. PMID- 11908345 TI - Degradation of gel in tray whitening. AB - This article will address the breakdown or lowering of active bleaching agent concentration during use in the mouth. A greater than expected degradation occurs during the first 5 minutes followed by an exponential rate loss. Subjects have a large variation in degradation rates. After 2 and 6 hours there was 52% and 24% respectively, of the active agent present in a carbamide peroxide gel. The presence or absence of the pellicle did not affect the rate of degradation. The relationship of gel degradation to the whitening capability of a gel is yet to be determined. PMID- 11908346 TI - Present and future technologies of tooth whitening. AB - Dental stains can be broadly classified as intrinsic or extrinsic. Intrinsic stains are a result of defects in tooth development, fluorosis, or acquired through the use of tetracycline. Extrinsic stains are localized mainly in the pellicle and are generated by the reaction between sugars and amino acids or acquired from the retention of exogenous chromophores in the pellicle. Three clinical methods are currently used for measuring stain removal and tooth whitening in the development of new whitening technologies: Lobene Stain Index, Shade Guide Color Change, and Minolta ChromaMeter. Professional tooth whitening products rely on proven technologies--35% hydrogen peroxide for in-office power bleaching or 10% to 15% carbamide peroxide for at-home bleaching--to reduce intrinsic stain and change the inherent tooth color. Over-the-counter tooth whitening products use a combination of surfactants, abrasives, anticalculus agents, and low levels of hydrogen peroxide to reduce extrinsic stain and help maintain tooth whiteness after professional treatment. Future technologies for whitening teeth could involve the use of activating agents to enhance the performance of hydrogen peroxide and natural enzymes. PMID- 11908347 TI - Peroxide-containing tooth whiteners: an update on safety. AB - The efficacy of peroxide-containing tooth whiteners has been well accepted; however, controversy concerning their safety has continued. Recently, a court ruling resulted in a ban on peroxide-containing tooth whiteners in the United Kingdom. While the decision was based on the classification of whiteners, there were issues with specious concerns about the safety, particularly carcinogenicity, of hydrogen peroxide (H2O2). This article reviews and discusses available scientific data on the carcinogenicity of H2O2 and consequently, their implications in the use of whiteners. An update on the effect of whiteners on the enamel surface is also included because controversial results were reported in recent studies. Based on current information, it is concluded that the proper use of dentist-monitored, at-home tooth whiteners containing 10% carbamide peroxide imposes no risk of carcinogenicity and does not cause irreversible damage to enamel. PMID- 11908348 TI - ADA guidelines for the acceptance of tooth-whitening products. AB - Since the introduction of at-home tooth whitening using peroxide-containing agents in 1989, the procedure has been well-received by the public and is becoming one of the most popular cosmetic dental procedures today. To ensure the safety and efficacy of this treatment, the Guidelines for the Acceptance of this category of products by the American Dental Association were developed. This article provides background on the American Dental Association (ADA) Seal of Acceptance Program. It also offers an overview of the ADA Acceptance Program Guidelines for Home-Use Tooth Whitening Products as well as the process through which the document was developed. PMID- 11908349 TI - Extraction sites--foundations for restorative excellence. AB - Preservation of the socket site architecture promotes better esthetics, easier prosthetic treatment, and restoration of the alveolar bone for implant or bridge replacement options. The purported simplicity of the simple extraction can be misleading if the bone housing or soft tissues have been ravaged by disease. This article provides guidelines for case selection, patient consultations, selection of appropriate materials, and a step-by-step methodology for completing successful extraction site therapy. PMID- 11908350 TI - The principles of uncomplicated exodontia: simple steps for safe extractions. AB - This article reviews the basic principles of patient evaluation and surgical techniques to accomplish extraction of teeth in an uncomplicated manner. Also presented are techniques for extraction-site grafting with bioactive glass. PMID- 11908351 TI - Ideal gingival form with computer-generated permanent healing abutments. AB - Implant fixture positioning and inclination problems that result from poor osseous height, width, and ridge configuration have created difficult prosthetic tooth replacement scenarios. Options for ameliorating poor angulation are limited to the use of preangled and custom abutments. Overcoming poor angulation has been simplified by using custom vs preangled abutments because preangled abutments are limited by their standardization to a few random angles. Custom abutments can be more predictably formed to re-create the desired supporting preparation orientation and morphology. This facilitates the formation of anatomical gingival topography and coronal contours for prosthetic replacement. Cast, ceramic, and machine-milled titanium abutments have several advantages and disadvantages. A new custom abutment system that uses computer-guided manufacturing technology to machine mill custom abutments from commercially pure titanium is described. These abutments are anatomically correct, have the proper emergence anatomy, proper spatial design at the cervical margins, necessary occlusal reduction, and the proper axial angulation of ideal tooth preparations. A clinical case that illustrates implementation, with respect to the principle of stage 2 guided tissue healing, is presented. PMID- 11908352 TI - Retrospective case series analysis of the factors determining immediate implant placement. AB - Implant placement at the time of extraction has become an acceptable treatment option. The formation of a restorative treatment plan frequently requires the removal of questionable and hopeless teeth. This retrospective case series analysis reports the reasons for tooth removal before immediate implant placement and provides a rationale for removing questionable and hopeless teeth. Root length is also analyzed as related to tooth loss. Between September 1986 and December 1998, 460 teeth were removed from 282 patients. Reasons for removal were advanced periodontal disease, endodontic complications, nonrestorable caries, roots fractures, short roots (< 14 mm in length), root resorption, and loosened posts. Implants were placed at the time of extraction. Tallies, frequency distributions, and percentages were used to determine individual and multiple reasons for extraction. For teeth with short roots, computerized measurements were made from periapical x-rays. Advanced periodontal disease and restored endodontically treated teeth with posts were the primary reasons for tooth extraction. Dental implants replaced 305 maxillary and 155 mandibular teeth. PMID- 11908353 TI - Antidepressants for chronic orofacial pain. AB - The management of chronic orofacial pain has a history of therapeutic misadventures, charismatic-based treatment philosophies, controversies over the correct nomenclature, and a lack of scientific documentation. The dental profession has struggled to develop a systematic approach to nomenclature, treatment, and clinical research through numerous conferences, workshops, and consensus attempts. Despite these efforts, there is no generally accepted agreement on the etiology of chronic orofacial pain, its natural history, the role of occlusion, the need for aggressive treatment, or the effectiveness, safety, and indications for most current practices. These professional differences are often fostered by a lack of appreciation for the difference between clinical observations, which may form the basis for therapeutic innovation, and the need to verify the safety and efficacy of treatments in studies that control for factors that can mimic clinical success. This article describes the use of antidepressants as an example for the treatment of chronic orofacial pain. The treatment arose from the clinical observations of astute clinicians, but was subjected to scientific validation in well-controlled clinical trials. PMID- 11908354 TI - Crown fractures: new concepts, materials, and techniques. AB - This article reviews the technology and science of crown fracture treatment as it applies to new materials, coupled with clinical research. To achieve clinical success, dentists need to embrace new scientific concepts and update the more traditional principles of therapy. Proposed treatments are outlined, which are based on new esthetic and insoluble adhesive materials and procedures that take into consideration the integrity of pulp biology. PMID- 11908355 TI - A new desensitizing dentifrice--an 8-week clinical investigation. AB - An 8-week, double-blind, three-way clinical trial compared the dentinal hypersensitivity-reducing effectiveness of a new dentifrice containing 5.0% potassium nitrate and 0.454% stannous fluoride in a silica base (Colgate Sensitive Maximum Strength Toothpaste, Colgate-Palmolive Co.) with a commercially available desensitizing dentifrice containing 5.0% potassium nitrate and 0.243% sodium fluoride in a silica base (Sensodyne Fresh Mint Toothpaste, Block Drug Company, Inc.) and a nondesensitizing dentifrice containing 0.243% sodium fluoride in a silica base (Colgate Winterfresh Gel, Colgate-Palmolive Co.). One hundred nine subjects were stratified into three balanced groups according to gender, age, mean baseline tactile (Yeaple Probe), and thermal (air blast) scores. The test products were randomly assigned to each group with instructions to brush twice daily. Oral examinations with tactile and thermal assessments were repeated after 4 and 8 weeks. The new dentifrice group demonstrated statistically significant improvements in tactile and thermal sensitivity over the two control groups. PMID- 11908356 TI - Comparative investigation of the desensitizing efficacy of a new dentifrice: a 14 day clinical study. AB - The effect on dentinal hypersensitivity from the use of a new formulation dentifrice containing 5.0% potassium nitrate and 0.454% stannous fluoride in a silica base (Colgate Sensitive Maximum Strength Toothpaste, Colgate-Palmolive Co.) over a 14-day period was compared to a commercially available, nondesensitizing control dentifrice (Colgate Winterfresh Gel, Colgate-Palmolive Co.). A total of 66 subjects were entered into and completed the study. They were stratified into two balanced groups according to their baseline mean tactile (Yeaple Probe) sensitivity scores and air blast (thermal) sensitivity scores. The two groups were randomly assigned to use either the new formulation dentifrice or the commercially available control dentifrice. Participants were instructed to brush their teeth twice daily (morning and evening) for 1 minute with their assigned dentifrice and a commercially available soft-bristled toothbrush. Dentinal hypersensitivity examinations (tactile and air blast sensitivity) were conducted at baseline and after 3, 7, 10, and 14 days' use of the products. All examinations were conducted by the same examiner. Participants who used the new formulation dentifrice containing potassium nitrate/stannous fluoride/silica demonstrated a statistically significant improvement in both tactile sensitivity and air blast sensitivity after 10 and 14 days' use of the dentifrice (p < 0.05), as compared to a commercially available control dentifrice. It was concluded that a new dentifrice formulation containing 5.0% potassium nitrate and 0.454% stannous fluoride in a silica base provided a statistically significant reduction in dentinal hypersensitivity after only 10 days, as compared to a commercially available nondesensitizing control dentifrice. PMID- 11908357 TI - Clinical study to compare extrinsic stain formation in subjects using three dentifrice formulations. AB - The purpose of this study was to compare the degree of extrinsic dental stain formed with the use of three dentifrices: (1) a new dentifrice formulation containing 5.0% potassium nitrate and 0.454% stannous fluoride in a silica base (Colgate Sensitive Maximum Strength Toothpaste, Colgate-Palmolive Co.); (2) a commercially available dentifrice containing 5.0% potassium nitrate and 0.243% sodium fluoride in a silica base (Sensodyne Fresh Mint Toothpaste, Block Drug Company, Inc.); and (3) a dentifrice containing 0.243% sodium fluoride in a silica base (Colgate Winterfresh Gel, Colgate-Palmolive Co.). A total of 121 participants were entered into the study and stratified into 3 balanced groups according to baseline mean Lobene Stain Index scores. A thorough dental prophylaxis was completed on each participant after completion of the baseline examination. The three groups were randomly assigned to use one of the three dentifrices. The groups were well balanced with regard to mean baseline stain index scores, gender, and tobacco habits. Participants were instructed to brush their teeth twice daily (morning and evening) for 1 minute with their assigned dentifrice and a commercially available soft-bristled toothbrush. Extrinsic dental stain examinations, which measured dental stain area and dental stain intensity, were conducted at baseline and 4 and 8 weeks. Examinations were conducted by the same dental examiner at each examination. After 4 and 8 weeks' use of the test dentifrices, there was no statistically significant difference regarding extrinsic stain formation with the use of any of the dentifrices. Thus, it can be concluded from this study that the use of a new formulation dentifrice containing 5.0% potassium nitrate and 0.454% stannous fluoride in a silica base will not result in a greater formation of extrinsic dental staining than that which is formed by two commercially available dentifrices not known to cause extrinsic dental stain. PMID- 11908359 TI - Local delivery of antimicrobials: a new era in the treatment of adult periodontitis. AB - This article discusses the principles, products, and techniques currently available for local delivery of antimicrobials in the treatment of adult periodontitis. Four principles provide the scientific basis for the treatment of periodontitis: it is caused by bacteria; it cannot be cured, but it can be controlled; clinicians cannot remove all the plaque and calculus; and periodontitis reinfects. This article stresses how the local delivery of antimicrobials can help the clinician achieve the goals of arresting the disease and maintaining the disease in the arrested or controlled state. Rationales for reevaluating the treated patient and treatment options are presented. Local delivery systems are reviewed, stressing those available in the United States. Pharmacokinetics, multicenter randomized trials, and techniques are presented. PMID- 11908358 TI - A randomized clinical trial of the desensitizing efficacy of three dentifrices. AB - The effect on dentinal hypersensitivity from the use of a new dentifrice containing 5.0% potassium nitrate and 0.454% stannous fluoride in a silica base (Colgate Sensitive Maximum Strength Toothpaste, Colgate-Palmolive Co.) over an 8 week period was compared to a commercially available dentifrice containing 5.0% potassium nitrate and 0.243% sodium fluoride in a silica base (positive control [Sensodyne Fresh Mint Toothpaste, Block Drug Company, Inc.]) and to a commercially available nondesensitizing dentifrice containing 0.243% sodium fluoride in a silica base (negative control [Colgate Winterfresh Gel, Colgate Palmolive Co.]). A total of 120 participants were stratified into 3 balanced groups according to baseline mean air blast (thermal) and tactile (Yeaple Probe) sensitivity scores, gender, and age. Participants brushed their teeth twice daily (morning and evening) for 1 minute. Dentinal hypersensitivity examinations were conducted at baseline, 4 weeks, and 8 weeks by the same dental examiner. After 4- and 8-weeks' use of their assigned products, participants in the new dentifrice group demonstrated statistically significant improvements (p < 0.05) in tactile and air blast sensitivity, as compared to those using the positive and negative control dentifrices. PMID- 11908360 TI - Rationale for the use of Atridox therapy for managing periodontal patients. AB - This article reviews the studies that provided the safety and efficacy data essential for the Food and Drug Administration approval of Atridox. These studies detail the clinical effectiveness of Atridox and provide the foundation for an understanding of the use of Atridox in the clinical management of patients with periodontitis. Atridox is a locally delivered, controlled-release system for the administration of high concentrations of doxycycline to the periodontal pocket. Nine-month clinical studies involving more than 800 patients have shown Atridox and scaling and root planing to be superior to placebo and oral hygiene for the efficacy parameters of attachment level, probing depth, and bleeding on probing. The positive clinical effects are consistent throughout a wide range of patients with varied histories of periodontal therapy. The implications of these findings as they relate to clinical practice are briefly addressed. PMID- 11908361 TI - Locally delivered doxycycline hyclate: case selection, preparation, and application. AB - This article presents the perspective of a private practice clinician who participated in the phase III clinical trials of Atridox (doxycycline hyclate) 10%. The selection of periodontal cases most likely to benefit from treatment with Atridox as part of overall comprehensive case management is discussed. Atridox may be used either before or after scaling and root planing or, in more rare circumstances, as a stand-alone therapy. Preparation of the material is discussed, and guidance for optimal application is provided. PMID- 11908362 TI - The supplemental use of antibiotics in periodontal therapy. AB - Many antibiotics and other chemotherapeutic agents have been used as adjuncts to mechanical periodontal therapy with mixed results. This article reviews the clinical and microbial results obtained after the systemic administration of tetracyclines, penicillins, clindamycin, and metronidazole, as well as the combination of metronidazole and Augmentin as adjuncts to conventional periodontal treatment. The major adverse effects associated with each of these antibiotics are given, as well as the potential for the emergence of antibiotic resistance in the periodontal flora. Recently, the introduction of a new generation of controlled-release, locally applied antimicrobial agents provides the clinician with the opportunity to treat individual periodontal sites with high concentrations of medication. The clinical effects obtained from multicenter clinical trials with PerioChip, which contains chlorhexidine, and with Atridox, which contains doxycycline, are summarized. Finally, suggestions are given both for the selection of an antimicrobial agent and for minimizing the development of antibiotic resistance in the periodontal flora. PMID- 11908363 TI - Current status of caries prevention. AB - The use of fluoride in water, professional topical applications, and dentifrices has resulted in a pronounced decline in dental caries. However, caries continues to afflict more 90% of the US population. Preventing and controlling dental caries requires the use of innovative measures to address the causes and reverse the caries process. This article provides an overview of the current strategies for the prevention of dental caries, the rationale for their use, and their relative efficacy. PMID- 11908364 TI - The relationship between periodontal disease and systemic conditions. AB - In this year's report of the United States Surgeon General on oral health in America, two major themes evolved: 1) oral health means much more than healthy teeth, and 2) oral health is integral to general health. This article describes how oral diseases, in particular periodontal diseases, are associated with other health problems, including cardiovascular disease, diabetes mellitus, complications of pregnancies, and osteoporosis. PMID- 11908365 TI - Genuine halitosis, pseudo-halitosis, and halitophobia: classification, diagnosis, and treatment. AB - Although tongue brushing and appropriate mouthrinses are both important and basic treatment measures for halitosis, other dental treatments are sometimes required. The treatment of genuine halitosis caused by oral conditions is not complex. In addition to genuine halitosis patients, psychosomatic halitosis patients also visit dental practitioners. Although psychosomatic halitosis is out of the treatment realm of dental practitioners, patients with this condition will still seek help from a dental practitioner. They often only receive treatment for genuine halitosis without referral to a psychological specialist. If these psychosomatic halitosis patients are incorrectly managed, the psychological condition might become worse than before the visit. To avoid the mismanagement of halitosis patients, classifications of halitosis patients have been established. Genuine halitosis was subclassified as physiologic halitosis and pathologic halitosis. Pathologic halitosis was further categorized to oral pathologic halitosis and extraoral pathologic halitosis. Both pseudo-halitosis and halitophobia patients complain of the existence of halitosis, which is not offensive. Pseudo-halitosis cannot be treated by dental practitioners, and halitophobia patients must be referred to psychological specialists. Clinicians need to examine the psychological condition of halitosis patients at the initial patient visit. A questionnaire prepared for the clinic at the University of British Columbia was found to be advantageous for this purpose. PMID- 11908366 TI - Oral precancer and cancer: etiology, clinical presentation, diagnosis, and management. AB - Oral and oropharyngeal cancers represent 3% of all cancers in the United States annually, with nearly 50% of people diagnosed with oral and oropharyngeal cancers dying as a result of the disease. Because the dental practitioner is in an ideal position for recognizing any abnormality of the oral mucosa, he or she is involved in the battle against oral cancer by helping establish the diagnosis at an early stage. This article presents the clinical appearance, explains the origins, and describes steps for the management of oral precancer and cancer. PMID- 11908367 TI - Assessment and management of periodontal infections: a medical-surgical approach. AB - Over the last 30 years, significant advances have been achieved in elucidating the etiology, pathogenesis, and treatment of periodontal diseases. A new paradigm has emerged that includes the initiation of disease by specific bacteria within a biofilm that stimulate an immunoinflammatory host response, resulting in host tissue destruction. Disease modifiers, which may be of genetic, environmental, or acquired origin, have been recognized as major determinants of disease severity and progression. Current treatment protocols include an assessment of risk factors to identify an individual's degree of susceptibility and therapeutic responsiveness. Basic and clinical research has resulted in the development of several strategies to identify specific bacteria and host-derived markers associated with disease progression. Preventive and therapeutic antimicrobial therapies, which use various delivery systems, have been devised to target drug placement to the infection site. More recently, host modulatory therapies have been created that inhibit disease progression through the reduction of inflammatory mediators. Finally, biological mediators, including growth and differentiation factors, have been used to enhance an individual's healing potential to achieve periodontal and bone regeneration. A combined medical surgical approach is indicated for these new methods of diagnosis, prevention, and treatment of periodontal diseases, which will allow for earlier treatment interventions and improved patient outcomes. PMID- 11908368 TI - Prosthodontic and restorative care in the 21st century. AB - In a recent report on oral health, Surgeon General Satcher reminded us of the need to change the attitudes of families concerning the importance of oral health. In prosthodontics and restorative dentistry, remarkable developments are occurring daily. Specialties are changing, and different types of dental practices are emerging. The introduction of computer technology has greatly affected the way the restorative dentist practices, and the evolution of cyberschools are no longer a fantasy. Adhesive dentistry has replaced the manner in which we prepare, restore, and bond restorations to teeth. The entire field of ceramics and methods of fabricating esthetic restorations are entering a new era. The exceptional prognosis of various implant systems has changed the way we think about maintaining hopeless teeth. Through tissue engineering, the 21st century may be revolutionary in the way we replace missing teeth and lost tooth structure. PMID- 11908369 TI - Thoughts on the future of dental and craniofacial research. PMID- 11908370 TI - Comparison of gingivitis and plaque efficacy of a battery-powered toothbrush and an ADA-provided manual toothbrush. AB - This examiner-blind clinical study evaluated the efficacy of a new battery powered toothbrush with oscillating head (Colgate Actibrush) on established gingivitis and plaque at 15 days and again at 30 days, as compared to a control manual toothbrush (American Dental Association [ADA]-provided toothbrush, full head, soft bristles). A total of 63 participants completed the study. They were stratified into two balanced groups according to their mean baseline prebrushing plaque scores and were randomly assigned to use the battery-powered test toothbrush or the manual control toothbrush. Participants were instructed to brush their teeth twice daily (mornings and evenings) for 1 minute with their assigned toothbrush for the 30-day duration of the study. Gingivitis and plaque (pre- and postbrushing) examinations were conducted by the same dental examiner at baseline, after 15 days, and again after 30 days. The Colgate Actibrush demonstrated a significantly greater reduction of plaque (46.53%) and gingivitis (18.57%) when compared to the ADA-provided toothbrush after 30 days of use. Additionally, a comparison of the plaque scores for the battery-powered toothbrush at 15 and 30 days shows a continued reduction in plaque of more than 25% for a cumulative difference from baseline of 73%. These results support the conclusion that the new battery-powered toothbrush is clinically superior in plaque removal efficacy and gingivitis efficacy to the manual toothbrush, and continues to significantly improve plaque scores even up to 30 days of use. PMID- 11908371 TI - Comparative efficacy of the Colgate Actibrush battery-powered toothbrush vs Oral B CrossAction toothbrush on established plaque and gingivitis: a 6-week clinical study. AB - The objective of this clinical program was to compare the efficacy of the Colgate Actibrush battery-powered toothbrush and the Oral-B CrossAction Toothbrush (full head, soft bristle) for the control of supragingival plaque and gingivitis. Two independent clinical studies were conducted: Study 1 (repeated 3 times) was a single-use, examiner-blind clinical study designed to measure the removal of plaque after 24 hours of no oral hygiene. Study 2 was a definitive 6-week, examiner-blind clinical study designed to determine plaque and gingivitis efficacy at 3 and 6 weeks. Sixty-one men and women, who had refrained from using oral hygiene procedures for 24 hours, were entered into the study and stratified into 2 balanced groups according to baseline (prebrushing) plaque and gingivitis scores. For Study 1, Modified Navy Plaque Index (Rustogi Refinement) scores were obtained prebrushing and after a 1-minute supervised brushing with the assigned toothbrush and a commercially available toothpaste. On 3 separate occasions, after 24 hours of no oral hygiene, the Colgate Actibrush battery-powered toothbrush removed significantly more plaque than did the CrossAction Toothbrush. For Study 2, subjects were instructed to brush their teeth twice daily for 1 minute with the assigned toothbrush. Plaque Index scores and Loe-Silness Gingival Index scores were assessed after 3 and 6 weeks. At the 6-week examination, the group using the Colgate Actibrush battery-powered toothbrush exhibited a statistically significant reduction in both supragingival plaque and gingivitis, compared with the group that used the CrossAction Toothbrush. The results of these clinical studies support the conclusion that the Colgate Actibrush battery powered toothbrush is clinically superior for the control of both supragingival plaque and gingivitis, as compared with the Oral-B CrossAction manual toothbrush. PMID- 11908372 TI - Effectiveness of a battery-powered toothbrush on plaque removal: comparison with four manual toothbrushes. AB - The objective of these four clinical studies was to evaluate the efficacy of the Colgate Actibrush battery-powered toothbrush in comparison with four top-selling manual European toothbrushes (Dr. Best Flex Toothbrush [Germany]; Reach Triple Action Ultra Clean Toothbrush [Germany]; Jordan Multi-Action Toothbrush [Norway]; and Sanogyl Systeme In'Side Double Action Toothbrush [France]) for plaque removal. Each clinical study used a single-use, examiner-blind design in which the Colgate Actibrush battery-powered toothbrush was compared with one of the 4 manual toothbrushes for removal of plaque in adult subjects after 24 hours of no oral hygiene. Supragingival plaque formation was assessed prebrushing and after a 1-minute supervised brushing with either the battery-powered toothbrush or 1 of the manual toothbrushes. After 24 hours of no oral hygiene, Colgate Actibrush removed statistically significantly more plaque than did all 4 of the manual toothbrushes. The results of these clinical studies support the conclusion that Colgate Actibrush is clinically superior for the control of supragingival plaque, compared with the leading selling European manual toothbrushes. PMID- 11908373 TI - Comparative efficacy of a new battery-powered toothbrush and an electric toothbrush on plaque removal. AB - This single-use, examiner-blind clinical study evaluated the plaque-removal efficacy of a new, battery-powered toothbrush (Colgate Actibrush) compared to a commercially available electric toothbrush (Braun Oral-B Plaque Remover) after 24 hours of no oral hygiene. Adult men and women reported to the clinical facility having refrained from oral hygiene procedures for 24 hours, and were stratified into 2 balanced groups according to plaque (prebrushing) scores. Participants then brushed their teeth for 1 minute, under supervision, with their assigned toothbrush and a commercially available toothpaste (Colgate Cavity Protection Great Regular Flavor Fluoride Toothpaste) and again were evaluated for supragingival plaque (postbrushing). The same dental examiner conducted the prebrushing and postbrushing plaque examinations. The study found no statistically significant difference in plaque removal between the group using the battery-powered toothbrush and the group using the electric toothbrush. Both power toothbrushes were found clinically to be equally effective with regard to the removal of 24-hour plaque accumulation. PMID- 11908374 TI - Comparative efficacy of Colgate Actibrush battery-powered toothbrush and Colgate Plus (manual) toothbrush on established plaque and gingivitis: a 30-day clinical study in New Jersey. AB - The objective of this 30-day clinical study, conducted in harmony with American Dental Association guidelines, was to evaluate the efficacy of a new battery powered toothbrush (Colgate Actibrush) relative to a manual toothbrush (Colgate Plus Diamond Head Toothbrush, Full Head, Soft Bristle) in the control of supragingival plaque and gingivitis. A total of 110 adult men and women from the Northern New Jersey area were entered into the study and stratified into 2 balanced groups according to baseline plaque and gingivitis scores. Participants were instructed to brush twice daily (morning and evening) for 1 minute with their assigned toothbrush and a commercially available toothpaste (Colgate Cavity Protection Great Regular Flavor Fluoride Toothpaste). Examinations for plaque and gingivitis were conducted by the same dental examiner at baseline, after 15 days, and again after 30 days of product use. All 110 participants complied with the protocol and completed the 30-day clinical study. At the 30-day examinations, the group using the Colgate Actibrush battery-powered toothbrush exhibited a statistically significant greater reduction in plaque (26.7%) and in gingivitis (25.8%) than did the group who used the Colgate Plus Diamond Head Toothbrush. The results of this 30-day clinical study support the conclusion that the Colgate Actibrush battery-powered toothbrush provides a clinically superior level of efficacy for the control of supragingival plaque and for the control of gingivitis when compared with a manual toothbrush. PMID- 11908375 TI - Comparative efficacy on supragingival plaque and gingivitis of a manual toothbrush (Colgate Plus) and a battery-powered toothbrush (Colgate Actibrush) over a 30-day period. AB - This clinical study compared the effect of Colgate Actibrush, a battery-powered toothbrush, and Colgate Plus Diamond Head, a full-head, soft-bristled manual toothbrush, on established supragingival plaque and gingivitis over a 30-day period. Sixty-two healthy adult men and women were entered in the study. The subjects were stratified into two balanced groups according to their mean baseline plaque and gingivitis scores. Each group was randomly assigned to the use of one of the two toothbrushes. Subjects were instructed to brush their teeth at home twice daily (morning and evening) for 1 minute with their assigned toothbrush and a commercially available tooth-paste for the 30-day duration of the study. Gingivitis and plaque examinations, as well as a soft-tissue evaluation, were conducted by the same dental examiner at baseline and after 15 and 30 days of toothbrush use. Plaque and gingivitis scores were reduced significantly from baseline after the use of both toothbrushes for 30 days. However, improvement in both supragingival plaque and gingivitis scores was significantly greater in the group using Colgate Actibrush. In conclusion, although both toothbrushes provided a significant plaque and gingivitis benefit when used as part of a normal oral hygiene regimen, the efficacy of the Colgate Actibrush was demonstrated to be superior to that of the Colgate Plus Toothbrush after 30 days of use. PMID- 11908376 TI - The direct posterior esthetic restoration using state-of-the-art composite resin technology. AB - As a result of the evolution of both materials and techniques, the direct posterior composite restoration has become a common procedure in today's dental practice. Advances in the adhesive protocol have allowed for the conservative preparation of the dentition by using the micromechanical potential of the sound tooth structure. Improvements of composite resin materials have further enabled the practitioner to re-create the natural esthetic beauty of the dentition while at the same time restoring the functional morphology. This article describes the technical protocol and materials necessary to perform the direct posterior composite restoration in the posterior dentition. PMID- 11908377 TI - Enhanced resilience and esthetics in a Class IV restoration. AB - The purpose of this article is to give the reader a better understanding of the complex restorative challenge in achieving true harmonization of the primary parameters in esthetics (i.e., color, shape, and texture) represented by the replacement of a single anterior tooth. The case presented demonstrates the restoration of a Class IV fracture taking esthetic consideration of the anatomic variations of the adjacent teeth to produce a direct composite resin in harmony with the surrounding dentition. The basic procedure includes tooth preparation, development of the body layer, internal characterization with tints, development of the artificial enamel layers, shaping and contouring, and polishing. In understanding the total morphology of a tooth and using natural teeth as the basis for morphological thinking, the clinician possesses the knowledge to create restorations with a more natural appearance. Use of a recently developed optimized-particle composite and this morphological thinking allowed the author to achieve a restoration in harmony with the surrounding dentition. Continuing technological breakthroughs allow the clinician to implement and maximize new products to attain more predictable and esthetic results as demonstrated by this methodological protocol of incremental application of composite resins and modifiers to transform the Class IV fracture into a final restoration that mimics nature. PMID- 11908378 TI - Achieving ultimate anterior esthetics with a new microhybrid composite. AB - Direct bonding is one of the most commonly used forms of restoration for the conservative esthetic improvement of anterior teeth. The major challenges involve selecting composites that have adequate strength as well as provide lifelike optical properties that render the restoration functionally sound and esthetically pleasing. Properties such as high sculptability, wide shade range, varying opacities and translucency, as well as high polishability are essential for gaining absolute control over the restorative process and the attainment of natural-looking results. This article discusses a new composite restorative system and its application in the direct esthetic and functional rehabilitation of the anterior dentition. PMID- 11908379 TI - Properties and characteristics of an indirect Bis-GMA/barium-glass polymer ceramic restorative system. AB - Although porcelain has predictably served the dental profession for more than 200 years, this class of restorative material has presented a number of clinically undesirable characteristics. As a result, during the past several years, major efforts have been made to enhance the mechanical and physical characteristics of polymers to better mimic porcelain for specific clinical applications. In recent years, Bis-GMA/barium-glass polymer systems have been developed to offer dental professionals a versatile restorative alternative to porcelain. This article details the material properties and clinical characteristics of one such system for the esthetic restoration of anterior and posterior teeth. PMID- 11908380 TI - Investigation of an adhesive system used with direct and indirect applications. AB - A laboratory investigation was performed that examined the adhesive properties of a new adhesive system. Contrasts were made with other commercially established systems, and indirect and direct bonding procedures were investigated. For the indirect evaluation, simulated restorations were bonded with an experimental resin cement. Sealing was assessed through microleakage analysis. Adhesive strength value was measured by shear bond strength methods. Further, the marginal quality of the restorations was assessed using scanning electron microscopy. Finally, the adhesive shear bond strengths were established using direct bonding. PMID- 11908381 TI - The myth of dentist-patient confidentiality. PMID- 11908382 TI - Two-year clinical evaluation of a high-density posterior restorative material. AB - The purpose of this study was to clinically evaluate SureFil high-density posterior composite for Class 2 restorations of permanent first and second molars. All teeth were restored in occlusion with the natural dentition. After cavity preparation, the enamel was etched for 15 seconds with 34% phosphoric acid. No liner/base material was used for the dentin. Prime & Bond 2.1 (two applications) was applied for adhesion to the exposed dentin and etched enamel. The first coat of adhesive was cured for 20 seconds with a curing light and the next coat was only spread thin with a gentle jet of air. The cavity was restored in 3-mm to 5-mm increments. Clinical evaluation (US Public Health Service method) and wear analysis (indirect cast comparison method) were conducted immediately after placement (baseline) and at 3 months, 6 months, 9 months, 1 year, and 2 years. A total of 25 restorations were inserted. At baseline and after 3, 6, and 9 months (n = 24), all restorations were graded Alfa in all categories. At the 1 year recall, 3 restorations (n = 22) were graded Bravo for surface staining. At the 2-year recall, an additional 3 restorations (n = 22) were graded Bravo for surface staining. No restoration was graded Charlie in any category at any time. Wear analysis revealed an average of 13.8 microns of wear at the 2-year recall (n = 22). No evidence of recurrent caries was observed. In conclusion, after 2 years, this high-density composite material demonstrated clinical acceptability and excellent wear characteristics in all categories, making it suitable for posterior restorations. PMID- 11908383 TI - Practical applications of the clinical microscope in a restorative dental practice. PMID- 11908384 TI - Strength of evidence linking oral conditions and systemic disease. AB - Associations between dental diseases and systemic outcomes are potentially important because of the high occurrence of dental diseases. If this extremely common source of chronic infection (dental disease) leads to an increased morbidity and mortality rate, the public health impact of oral disease on millions of Americans would be substantial. Recent studies demonstrate an association between dental and systemic diseases, including systemic infections, cardiovascular disease, pregnancy outcomes, respiratory diseases, and increased all-cause mortality rate. Because there are several common risk factors for oral and systemic diseases, and limitations in published studies, a careful interpretation is needed. Confounding (shared risk factors for both systemic and dental disease) may explain part of the reported associations. It is also plausible that there may be a causal link. It is likely that if there is a causal link, several pathways and mediators coexist, linking oral and systemic disease. Bacteremia, bacterial endotoxins, cytokines, and other inflammatory mediators could conceivably be playing a direct or indirect role. Missing teeth are a surrogate marker for previous dental infection, and may also lead to altered dietary intake. Hence, diet may be an additional mediator for several of these outcomes. We caution clinicians not to recommend extracting infected teeth, based on the periodontal-systemic disease associations, if the teeth do not warrant extraction otherwise, because loss of teeth and edentulousness are associated with increased risk of systemic diseases. When assessed against causal-defined criteria, the evidence suggests possible causal associations between chronic periodontal disease and tooth loss with cardiovascular disease, bacterial endocarditis, pregnancy outcomes, and all-cause overall mortality. Further studies are needed to show consistency, to corroborate that the associations are independent of common risk factors for both systemic and dental disease, including healthy lifestyle factors, and to evaluate different biological pathways. PMID- 11908385 TI - Influence of behavioral science research on oral health promotion. AB - To realize the benefits of advances in preventive and problem-focused dentistry, patients must carry out specific oral health behaviors. The behaviors range from seeking preventive care, to keeping appointments, to carrying out home-based self care regimens. Unfortunately, rates of these behaviors are less than optimal and maximizing patient involvement in dental care is a major challenge facing dentistry. In fact, poor rates of patient adherence remains a significant problem in all areas of health care. Aspects of the treatment regimen itself as well as provider and patient actions impact these outcomes. Theoretical models grounded in the behavioral sciences provide useful frameworks for understanding the process of health behavior change that can be applied to patient care. Research in health promotion and disease prevention have identified a number of psychological, social, and environmental issues related to oral health behaviors and outcomes, including personal barriers, social support, stress and coping, health beliefs, and dental anxiety. These barriers are discussed, and examples of interventions that have incorporated theoretical models and behavioral outcomes and linking them to oral health are provided. Future directions of the expanding area of behavioral science and oral health promotion in clinical practice are discussed. PMID- 11908386 TI - Smoking and stress: common denominators for periodontal disease, heart disease, and diabetes mellitus. AB - The scientific evidence supporting the relationship between periodontal disease and two major health problems of the US population, i.e., cardiovascular disease and diabetes mellitus, is discussed. Evidence supporting the behavioral and psychosocial factors mediating the relationship between periodontal infection and these systemic conditions, namely smoking and stress, is presented, as well as models explaining these relationships. Treatment protocols are suggested for the clinical management of patients who smoke, are diabetic, or are at risk for cardiovascular disease. PMID- 11908387 TI - The intersection of socioeconomic variables, oral health, and systemic disease: all health care is cultural. AB - In the last decade, there has been an increase in the proportion of the American population that lacks health insurance. Additionally, long-term disparities in health access and status for populations of color have continued to exist without major change. A review of the literature in periodontia shows a statistical relationship between the risk of periodontal disease and a range of sociodemographic characteristics (race, income, social class). However, the factors giving rise to this increased risk or how to reduce it are unclear. Cultural competence, an elaboration of biopsychosocial theory, hypothesizes a relationship between the culture of a group and its risk of disease and the kind and quality of services obtained. Four areas of a culturally competent approach are cited as a minimum base of standards for providing services to people of color. PMID- 11908388 TI - Influences of personality and coping variables on health behaviors and health outcomes. AB - Many health problems begin with behavior: taking risks or failing to take proper precautions. Psychosocial processes of several types underlie such behaviors. It follows that research on health can benefit from considering psychosocial processes as well as biological ones. This article focuses on two general domains of psychological influences on health--personality and coping. Several approaches to the domain of personality are described, along with several issues involved in the conceptualization of coping. Examples of research on how these phenomena relate to health behaviors and health outcomes are presented. PMID- 11908389 TI - Preterm birth, osteoporosis, and periodontal disease. AB - The purpose of this two-part article is to review two major events in the life span of a woman. These include the putative relationship between oral health, pregnancy, and postmenopausal osteoporosis. Current knowledge about risk factors for preterm birth and for osteoporosis are discussed. The newest studies that address the relationship between oral and systemic health are also reviewed. PMID- 11908390 TI - The Oral Risk Assessment and Early Intervention System--a clinician's tool for integrating the bio/psycho/social risk into oral disease interventions. AB - An association between oral diseases and systemic diseases has been suspected for centuries. More recently, investigation has provided insight into the impact of psychologic factors, social value systems, beliefs about disease processes and predictive treatment outcomes. Historically, dentistry has focused primarily on the restoration of physical function rather than broader multifactorial issues that impact long-term health maintenance. Dental patients are a complex interaction of biologic susceptibilities, lifestyle behaviors, values, and disease risk variables, which require an interdisciplinary approach to provide optimal health care. This article provides information-processing tools to assist clinicians in developing appropriate patient interventions by employing on Oral Risk Assessment and Early Intervention System. PMID- 11908392 TI - A review of the curing mechanics of composites and their significance in dental applications. AB - Much of the technique sensitivity associated with polymer matrix composites is a direct result of their curing shrinkage. Challenges with marginal integrity, adaptation of proximal contact, and residual stress are related to this intrinsic property. There are many test methods described in the literature that measure various aspects of polymerization contraction. Some measure total contraction, which is the sum of pre- and postgelation shrinkage, whereas others are sensitive only to postgelation deformation, which occurs after the onset of measurable stiffness. Development of methods to compensate for curing shrinkage is best described on the basis of an understanding of the polymerization mechanics. The distinction between total and postgelation contraction, and recognition of limitations of test methods are important considerations when interpreting literature data before selecting a restorative material. PMID- 11908393 TI - Theoretical considerations of contraction stress. AB - Polymerization shrinkage of composite restorative materials causes clinical concerns because it introduces residual stresses in restored teeth. These stresses can, among other things, propagate enamel cracks, bring about microleakage, and cause postoperative sensitivity. The amount of shrinkage stress does not depend only on how much a composite contracts, but also on the elastic modulus ("stiffness") of the composite, the shape of the cavity, the established bonding between the tooth and restoration, etc. The relationships between these many factors can be described by universal physical laws. To analyze shrinkage stresses, theoretical models must be used that relate the various shrinkage properties and clinical conditions via physical laws. Models will assist us to rethink and optimize established paths to clinical success. PMID- 11908394 TI - Controlling and understanding the polymerization shrinkage-induced stresses in light-cured composites. AB - The shrinkage of composites induces stress in the final restoration when the composite is bonded to the tooth surface. The amount of stress can be controlled by the method of pulse-delay cure used. The development of new composites has changed the energy requirements for obtaining polymerization. The total energy required for optimal polymerization has been reduced and the rate at which the energy is delivered can have a strong effect on the final properties of the restoration. Newer composites may employ different photoinitiators, making the spectral emission from the curing light an important factor for properties of the final restoration. It is proposed that labels for composites carry: (1) indications for the total energy required; (2) specification of the pulse-delay cure if applicable; and (3) the required spectral bandwidth of the curing lamp light emission. PMID- 11908395 TI - Contemporary issues in photocuring. AB - Currently, there is much confusion regarding differences among the variety of light-curing units on the market. Units range from conventional, fixed, continuous output quartz-tungsten-halogen (QTH) models, to very high-intensity systems such as the plasma-arc (PAC) units and the argon-ion lasers. Also, a trend has been established for purposefully providing low light intensity output during initial phases of the curing exposure: the so-called "soft-start" method. Proponents of this method state that by decreasing the rate at which a photoactivated resin cures, stress buildup at the tooth/restoration interface can be minimized, resulting in enhanced marginal integrity. PMID- 11908396 TI - Resin polymerization problems--are they caused by resin curing lights, resin formulations, or both? AB - Negative effects of rapid, high-intensity resin curing have been predicted for both argon lasers and plasma-arc curing lights. To address these questions, six different resin restorative materials were cured with 14 different resin curing lights representing differences in intensities ranging from 400 mW/cm2 to 1,900 mW/cm2; delivery modes using constant, ramped, and stepped methods; cure times ranging from 1 second to 40 seconds; and spot sizes of 6.7 mm to 10.9 mm. Two lasers, five plasma-arc lights, and seven halogen lights were used. Shrinkage, modulus, heat generation, strain, and physical changes on the teeth and resins during strain testing were documented. Results showed effects associated with lights were not statistically significant, but resin formulation was highly significant. Microfill resins had the least shrinkage and the lowest modulus. An autocure resin had shrinkage and modulus as high as or higher than the light cured hybrid resins. Lasers and plasma-arc lights produced the highest heat increases on the surface (up to 21 degrees C) and within the resin restorations (up to 14 degrees C), and the halogen lights produced the most heat within the pulp chamber (up to 2 degrees C). Strain within the tooth was least with Heliomolar and greatest with Z100 Restorative and BISFIL II autocure resin. Clinical effects of strain relief were evident as white lines at the tooth-resin interface and cracks in enamel adjacent to the margins. This work implicates resin formulation, rather than light type or curing mode, as the important factor in polymerization problems. Lower light intensity and use of ramped and stepped curing modes did not provide significant lowering of shrinkage, modulus, or strain, and did not prevent enamel cracking adjacent to margins and formation of "white line" defects at the margins. Until materials with lower shrinkage and modulus are available, use of low-viscosity surface sealants as a final step in resin placement is suggested to seal defects. PMID- 11908397 TI - Operation and diagnostic features of the VIP light. AB - Composite curing lights have been an equipment staple in the dental practitioner's office for many years. This article offers some insights into the design considerations of quartz-tungsten-halogen dental curing lights as well as the peripheral devices associated with these lights. Some of the features of the new Bisco VIP light are explained. PMID- 11908399 TI - Clinical experience with PYRAMID stratified aggregate restorative and the VIP unit. AB - The shrinkage of composites induces stress in the final restoration when the composite is bonded to the tooth surface. The amount of stress can be controlled by the method of pulse-delay cure used. The development of new composites has changed the energy requirements for obtaining polymerization. The total energy required for optimal polymerization has been reduced, and the rate at which the energy is delivered can have a strong effect on the final properties of the restoration. Newer composites may employ different photoinitiators, making the spectral emission from the curing light an important factor for properties of the final restoration. It is proposed that labels for composites carry: (1) indications for the total energy required; (2) specification of the pulse-delay cure if applicable; and (3) the required spectral bandwidth of the curing lamp light emission. PMID- 11908398 TI - In vitro wear of packable dental composites. AB - The in vitro abrasion and attrition wear of several packable composites were evaluated on the Oregon Health Sciences University oral wear simulator and compared to two popular commercial composites. In general, the wear of the packable composites was equivalent to that of the nonpackable microfill and minifill composites. The wear of the enamel cusps opposing the composites in the wear machine was also found to be similar to that of enamel opposing the nonpackable composites. Based on these results, one would expect that these new packable composites would not offer improved clinical performance over current nonpackable composites in terms of wear resistance. PMID- 11908400 TI - The cariostatic mechanism of fluoride. AB - The benefits of using fluoride to prevent caries have been known for many years, but a complete understanding of this mechanism is still being researched. The fluoride concentration in the apatitic structure of enamel does not have as significant an effect on reducing caries as a continuous presence of fluoride in the plaque liquid. Concentrated, topical fluoride agents (such as in toothpaste, fluoride mouth rinses, gels, or varnishes) have a different mechanism of fluoride protection than low-concentration applications (such as fluoridated water). In initial caries lesions and plaque, concentrated agents form globules of a calcium fluoride-like material on the enamel surface. This material is fairly insoluble, possibly because it is coated with phosphates or proteins. This mechanism explains how the topical application of a fluoride varnish, two or three times a year, can result in caries reduction. PMID- 11908401 TI - Efficacy and safety of fluoride varnishes. AB - Clinical studies show that fluoride varnishes, such as Duraphat, are effective in increasing the fluoride content in the enamel and preventing caries. Varnish application is fast and easy. A professional prophylaxis before varnish application is not necessary, which decreases the application time. Patients receive significant preventive benefits with only semiannual varnish applications. Finally, and most importantly, studies show that fluoride varnishes are safe. Effectiveness, application ease, and safety give fluoride varnishes an advantage over other types of topical fluoride treatments, such as gels and rinses. PMID- 11908402 TI - Treating hypersensitivity with fluoride varnish. AB - Dentinal hypersensitivity results when stimulation causes the fluid in open dentinal tubules to undergo pressure changes, which activates mechanoreceptor nerves and results in pain. Treatment with fluoride varnish forms a protective layer of calcium fluoride that prevents this fluid flow, thereby reducing dentinal sensitivity. PMID- 11908403 TI - Evolution of professionally applied topical fluoride therapies. AB - Ever since researchers first realized the benefits of fluoride as an anticaries agent, they began investigating the effect of concentrated topical fluoride applications in reducing caries. Many forms of fluoride were studied, including solutions, gels or foams of sodium fluoride, stannous fluoride, organic amine fluoride, acidulated phosphate fluoride, and nonaqueous fluoride varnishes in an alcoholic solution of natural resins and difluorosilane agents covered by a polyurethane coating. All of these professionally applied topical agents have anticaries benefits, although these benefits and the ease of application vary. One trend that stands out in the evolution of professionally applied fluoride agents is the development of increasingly more user-friendly products. PMID- 11908404 TI - Vital tooth whitening with a novel hydrogen peroxide strip system: design, kinetics, and clinical response. AB - For many years, at-home whitening has been used with great success and produces some of the most satisfying results of all dental procedures. Historically, the most common procedure used was a custom-fabricated tray loaded with a 10% carbamide peroxide gel that was worn overnight. Today, many manufacturers offer higher concentrations (15% and 20% carbamide peroxide) for faster results. Regardless of the peroxide concentration used, the custom tray delivery system has remained essentially the same. Recently, a trayless whitening system was developed that does not require any prefabrication or gel loading. The new delivery system is a thin, conformable strip precoated with an adhesive hydrogen peroxide gel. Each preloaded strip is presented on a backing liner. To use the strip, it is peeled off of the backing liner and applied to the facial surfaces of the anterior teeth. Each strip is worn for 30 minutes, removed, and discarded. The strip holds the gel in place for sufficient time to allow the peroxide to intrinsically and extrinsically whiten the teeth. The highly flexible strips conform intimately to the tooth surface and provide a uniform, controlled application of the peroxide gel. PMID- 11908405 TI - Tooth-whitening efficacy and safety: a randomized and controlled clinical trial. AB - A randomized, single-center, double-blind, parallel-group, placebo-controlled trial evaluated the whitening efficacy and safety of 2-week, twice-daily use of a 5.3% hydrogen peroxide tooth-bleaching gel delivered on polyethylene film. Efficacy was based on change in Vita shade scores from baseline to the end of treatment. Thirty-three patients in each group completed treatment. Use of the peroxide-containing gel led to a mean change in baseline Vita shade score of 3.70 +/- 0.35, compared with a change of -0.87 +/- 0.24 after use of a placebo gel. After adjustment for baseline scores, the mean difference in shade change between the peroxide gel-treated group and placebo-treated group was -2.85 +/- 0.41 (P < 0.0001). Both treatments were generally well tolerated. The strips offer ease of use, comfort, and shorter duration of wear compared with other at home bleaching systems. PMID- 11908406 TI - A randomized clinical trial comparing a novel 5.3% hydrogen peroxide whitening strip to 10%, 15%, and 20% carbamide peroxide tray-based bleaching systems. AB - A randomized and controlled, parallel-group clinical trial compared the whitening benefits of a novel 5.3% hydrogen peroxide bleaching strip to 10%, 15%, and 20% carbamide peroxide tray-based bleaching systems. A total of 36 healthy adults were randomized to a 14-day regimen in which both arches were whitened for 1 hour per day in the whitening-strip group or 2 hours per day in the tray groups. Efficacy was measured objectively using digital images of the anterior teeth at baseline and after 14 days of treatment. Overall tooth color (L*a*b*) was derived from individual pixel values, and then mean levels of delta b*, delta L*, and composite color (delta E*) were compared using analysis of covariance. After 14 days of treatment, all groups experienced a greater than 1-unit mean improvement in delta b*, delta L*, and delta E* relative to baseline. For the primary study variable, reduction of yellow (delta b*) outcomes after 14 hours of using the experimental strip were comparable to those observed with the 10% tray group after 28 hours of use. These two treatment groups did not differ statistically with respect to any of the color measurements used in this study. For the tray groups, there was a reasonable dose relationship for the primary end point, delta b*, with the 15% and 20% tray groups averaging 17% and 68% improvements in yellow, respectively, over the 10% group. Except for the 20% carbamide peroxide system, where sensitivity was relatively common, all test products were well tolerated. In this first comparative evaluation vs marketed controls, use of the whitening strips twice daily for 14 days yielded a highly significant improvement in tooth color vs baseline. PMID- 11908407 TI - Effects of tooth-whitening gels on enamel and dentin ultrastructure--a confocal laser scanning microscopy pilot study. AB - Confocal laser scanning microscopy (CLSM) in reflection mode provides a useful means for nondestructive microscopic examination of the ultrastructural characteristics of hard tissues, including enamel and dentin. In this study, CLSM was used to examine the effects of in vitro bleaching on enamel and dentin. The crowns of extracted human third molars were sectioned below the occlusal fissure, revealing subsurface dentin and the outer surface ring of enamel. Specimens were polished with 1,200-grit alumina followed by fine polish lapidary film, and cut into four equal sections per tooth, allowing each tooth to serve as internal control. Sections were mounted in acrylate for handling and were bleached for 0, 15, and 30 hours in 0.25 g of commercial Opalescence whitening gel (10% carbamide peroxide), the 5.3% hydrogen peroxide gel used in Crest Whitestrips, or 5% HClO4 solution. Blank glycerin served as a control treatment. In vitro whitening was confirmed by colorimeter readings of delta b and delta L with a PR 650 spectrophotometer/colorimeter. Treated teeth were examined with CLSM comparing enamel surface, dentoenamel junction (DEJ), and dentin at approximately 5 microns subsurface to the polished surfaces under an oil immersion objective. A single sample from each treatment (30-hour whitening) was also examined by environmental variable pressure scanning electron microscopy (VP-SEM). Internal comparison of glycerin (no whitening) controls (0-, 15-, and 30-hour exposures) revealed reproducible ultrastructure, facilitating treatment comparisons. Whitened teeth revealed no significant micromorphological changes associated with the whitening process in subsurface enamel, DEJ, and dentin areas. The direct treatment of cross sections permitted simple access of all areas to whitening gel, thereby eliminating the possibility of diffusion limitation of the gel in producing artifacts. VP-SEM observations similarly showed no significant changes in the surfaces after treatment. PMID- 11908409 TI - Shifting paradigms in whitening: introduction of a novel system for vital tooth bleaching. AB - The advent of whitening strips (Crest Whitestrips) affords a novel system for peroxide delivery without custom tray fabrication. In addition to generalized bleaching indications, clinical applications may leverage the low overall dose and short wearing regimen with the easy-to-use strips. The impact of these strips on the dental practice may be direct, as a result of increased accessibility via professional or personal use, or indirect, because of advertising and research in the area of tooth whitening. PMID- 11908408 TI - Impact of demographic, behavioral, and dental care utilization parameters on tooth color and personal satisfaction. AB - A cross-sectional survey across broad age ranges was conducted to evaluate demographic, behavioral, and treatment parameters that impact tooth color and its perception. The sample included 180 US adults and teenagers, with a comparable representation of males and females in 6 different age strata, ranging from 13 to 64 years. Tooth color (L*a*b*) was measured on the maxillary central incisors using a spectrophotometer, and first-person satisfaction with tooth color was assessed using a five-point qualitative scale. Demographic, behavioral, and oral care parameters were modeled using multiple regression analysis. After adjusting for other explanatory variables, age, gender, coffee/tea consumption, and dental care all significantly affected yellowing (b*) and brightness (L*). Dental-visit frequency was the only factor that significantly predicted self-satisfaction with tooth color, explaining just 3% of the overall variability. First-person dissatisfaction with tooth color was common and found in most demographic and behavioral cohorts. Although age contributed to objectively measured tooth discoloration, personal satisfaction with tooth color was age-independent. These results suggest that the need or demand for esthetic dentistry may be broad-based and transcend stereotypical perceptions. PMID- 11908410 TI - A prospective clinical evaluation of electronically mixed polyvinyl siloxane impression materials: results from the prosthetic "SuperStudy"--a consumer evaluation. AB - Polyvinyl siloxane (PVS) impression materials incorporating a polyether carbosilane wetting agent and mixed with an electronic mixing system (Pentamix) were clinically compared with traditionally delivered (i.e., automixed) PVS impression materials during routine use by 1,505 general and specialized dental practitioners evaluating more than 30,000 impressions. Each study participant selected 20 patients and used standard tooth-preparation procedures appropriate to the therapy required, supplied specific data on each case, and ultimately evaluated the marginal detail, fit, and success of the final restorations. The areas requiring evaluation upon completion of the final restorations were ranked between "excellent," "good," "poor," and "remake needed," among users of the PVS materials with the electronic mixing system. About 80% of the respondents rated the Pentamix system as easier to mix and deliver than the gun or hand-mixed systems and two thirds said it was faster to mix. The system also received high scores for hygienic delivery, ease of mixing, and clean-up. PMID- 11908411 TI - The effect of the pulse-delay cure technique on residual strain in composites. AB - Polymerization-induced shrinkage of composites leads to residual stress in the final restoration. For composites with a high modulus of elasticity, the level of stress can have significant clinical consequences, including crack formation in the enamel or microscopic separations at the preparation/restoration interface. The pulse-delay cure technique cures composites by providing low-energy pulse initially (e.g., 200 mW/cm2 for 3 seconds), followed by a waiting period of 3 to 5 minutes for strain relief, during which the composite can be finished and polished. The final cure is obtained by exposure to a high-intensity light source of 500 mW/cm2 for the recommended time. In vitro data obtained by strain gauges show that the pulse-delay cure technique can reduce residual strain in the composite by as much as 34%. PMID- 11908414 TI - [Acute anterior instability of the shoulder: short- and mid-term outcome after conservative treatment]. PMID- 11908413 TI - Adjustable gastric and esophagogastric banding. Is a pouch compulsory? PMID- 11908415 TI - [Nosocomial infections in orthopedic surgery. Round table of the 75th meeting of the French Society of Orthopedic Surgery and Traumatology, 7 November 2000]. PMID- 11908416 TI - Assessing the present and future roles of infectious diseases specialists in treating infections. PMID- 11908417 TI - Structure of the porcine LH- and FSH-releasing hormone. I. The proposed amino acid sequence. 1991. PMID- 11908418 TI - A new plastic operation for stricture at the uretero-pelvic junction. Report of 20 operations. 1937. PMID- 11908419 TI - Anatrophic nephrotomy and plastic calyrhaphy. 1968. PMID- 11908420 TI - Transurethral prostatectomy: immediate and postoperative complications. a cooperative study of 13 participating institutions evaluating 3,885 patients. 1989. PMID- 11908421 TI - [Citric acid as an adjunct to periodontal surgery]. AB - This review focuses on the effect of acid demineralisation of the root surface as an adjunct to periodontal surgery in order to obtain new attachment. Several animal studies have shown that the treatment facilitates attachment of connective tissue to the root surface. Clinical studies, however, have failed to demonstrate the effectiveness of citric acid as an adjuvant to surgical debridement. PMID- 11908422 TI - [Chewing: a matter of selection and breakage]. AB - The masticatory performance of subjects can objectively be determined by describing the particle size distribution of a comminuted test food (cubes Optosil). After a fixed number of chewing strokes the particles are collected and sorted with a stack of sieves. The size distribution can be characterized with two parameters; the median particle size and the broadness of the distribution. The rate of reduction of the median size is used to characterize the chewing performance. The chewing process can be described as the result of two processes: selection and breakage. The parameters of these two processes can be derived from the chewing result using a mathematical model. The differences in chewing performance between subjects with a natural dentition and subjects with a complete denture occurs already in the first 20 chewing strokes. Experimental results indicate that especially the selection chance is smaller in denture wearers. PMID- 11908423 TI - [Adhesive dentistry II. Direct techniques, practical course]. PMID- 11908424 TI - [(Neo)natal tooth inperspective. Literature review and report of two cases]. AB - The 'problem' of the (neo)natal tooth is discussed in conjunction with a report of two cases and a review of the literature. The estimated incidence is between 1:700 and 1:3500 births. Girls are more often affected than boys. It usually concerns the two lower central deciduous incisors. The cause is most likely to be a superficial position of the tooth-germ, in which a hereditary component is a positive factor. The tooth is often deficient and very mobile, which may require treatment. In these cases extraction is the best therapy, if necessary after administration of vitamin K. PMID- 11908425 TI - [Application of titanium-nitride coatings in dentistry]. AB - Titanium and its alloys are increasingly important in dentistry. Thin titanium nitride (Ti-N) coatings improve the properties of metallic material for industrial purposes. Recently Ti-N coatings have been advocated in dentistry. In a survey of literature aspects of biological, mechanical and corrosive behaviour are assessed. Moreover, specific problems for clinical application are indicated. It is concluded that as long as the integrity of the Ti-N coating is not guaranteed under critical conditions one should be cautious with its clinical application. By no means it can be used for improvement of deficient dental alloys. PMID- 11908426 TI - [Dental hygienist in financial independent practice: an established institute?]. AB - The purpose of this article was to describe the development and actual status of the financial independent dental hygienist in The Netherlands. In 15 years nearly 100 dental hygienists started in financial independent practice after a few years of experience. In a survey (response: 80%, N = 58) respondents reported that especially patients with advanced periodontal diseases were treated in their practice. A majority of the respondents also works for a dentist employer. Compensation by insurance companies makes the treatment by independent dental hygienists approachable for more patients and is therefore advisable. PMID- 11908428 TI - [X-ray diagnosis. Introduction, incorporation and exploration of a new technique]. PMID- 11908427 TI - [Observing the tooth. The paths of light in visual and instrumental observation]. AB - The light we observe when we look at a tooth has travelled a long way through the tooth. A large fraction has passed through the dentine. There is also a sideways displacement: the point of entrance into the tooth is about 2 mm away from the point of exit. This implies that tooth color cannot correctly be measured with an instrument employing a single circular window for both illumination and measurement. Visual caries diagnosis of smooth surfaces and with Fiber Optic Trans Illumination (FOTI) is discussed in terms of light paths through the tooth. PMID- 11908429 TI - [Dental fillings]. PMID- 11908430 TI - [A bird's eye view of the development of periodontics in the Netherlands. From pyorrhea alveolaris to periodontal disease]. PMID- 11908431 TI - [A hundred years of dental practice hygiene]. PMID- 11908432 TI - [Fluoride]. PMID- 11908433 TI - [Oral hygiene in the next century]. PMID- 11908434 TI - [The echo of Angle]. PMID- 11908435 TI - [The development of gnathology: from occlusions to function]. PMID- 11908436 TI - [Crowns and partial dentures currently and in the past]. PMID- 11908437 TI - [Is there still room for partial dentures?]. PMID- 11908438 TI - [From dentures to partial dentures]. PMID- 11908439 TI - [History of maxillofacial prosthetics]. PMID- 11908440 TI - [Mouth diseases and jaw surgery. Between past and future]. PMID- 11908441 TI - [Re-, trans- and implantation]. PMID- 11908442 TI - [A hundred years of 'social dentistry' thinking]. PMID- 11908443 TI - [Jurisprudence?]. PMID- 11908444 TI - [Foreword]. PMID- 11908445 TI - [The jubilee year of literary colleagues]. PMID- 11908447 TI - [From endodontic disease to endodontics]. PMID- 11908446 TI - [From preventive dentistry to dental caries]. PMID- 11908448 TI - [Peripheral facial nerve palsy]. AB - There are different etiological factors concerning the acute peripheral facial nerve palsy. In the majority of the cases, however, no etiological factor can be found. These cases are called idiopathic facial palsy or Bells palsy. Perhaps local anaesthetics could play a role as an etiological factor. By means of a case report this form of facial nerve palsy will be discussed. PMID- 11908449 TI - [Prevention of bacterial endocarditis. Revision of guidelines for endocarditis prevention 1992]. PMID- 11908450 TI - [Revised guidelines for bacterial endocarditis prophylaxis]. PMID- 11908451 TI - [Ethnicity]. PMID- 11908452 TI - [Turkish and Moroccan migrants in the Netherlands. Some background information]. AB - During the years 1960-1970 a large number of foreign workers from mainly Turkey and Morocco came to the Netherlands. Most of the men came from rural areas and were poorly educated. In the seventies wives and children came to join their husbands and fathers. A lot of ths 'first generation' of men and women are now unemployed. Bad health makes it impossible for them to work. Their children and grandchildren are growing up in the Netherlands and quite a number of them are not benefitting fully from education and as a consequence do not have good opportunities in the job market. The legal circumstances for foreigners are very complicated. The many changes in their legal status create a feeling of continuous insecurity about their residence here. Most of the migrants have a low social-economic status and as such are at risk with their health. The older migrants mainly wish to return to their own country, but feel that they are obliged to stay with their children in the Netherlands. Most of the migrants came to the Netherlands in order to work, earn what is to them a substantial income, return home with their savings and start a business or settle in their own country. Their current situation in the Netherlands makes it very difficult for them to attain this ideal. PMID- 11908453 TI - [MRI and MRS diagnosis. Application in multiple sclerosis patients and research animals with experimental allergic encephalomyelitis]. AB - MR imaging and spectroscopy offer a noninvasive way to observe lesions and biochemical changes, respectively, that may provide new insights into demyelinating diseases such as MS. Although the role and importance of some of the metabolites, such as choline and N-acetyl aspartate in brain function and disease are not fully understood, the specificity of these changes may provide information about the stage and reversibility of the brain lesions. Animal studies are particularly insightful when MR spectroscopic signals are correlated with histologic and biochemical techniques. Clinical application of spectroscopic imaging could arise from the capability to differentiate between early lesions that might respond to therapy and older irreversible lesions. Although more data need to be obtained, MR spectroscopy shows promise for monitoring progression of MS and evaluating therapy. PMID- 11908454 TI - [Determination of vertical dimension of dental occlusion]. AB - Principles and concepts in determining the vertical dimension of occlusion in mutilated dentitions are summarized. Historically, most concepts have been developed for full dentures. However, in the rehabilitation of a mutilated dentition it is often necessary to increase or change the vertical dimension of occlusion. If the vertical dimension of occlusion has to be altered the patient's adaptability to the new position should be tested during a trial period. Determination of the vertical dimension of occlusion is apparently a combination of art, science and experience. PMID- 11908455 TI - [A protocol for the treatment of periodontal disease]. AB - The importance of a number of clinical indicators for the diagnosis, the treatment and the evaluation of periodontal disease is discussed. PMID- 11908456 TI - [A.a.: the bacterium with the long name]. AB - The introduction of Kodak's chairside test kit, Evalusite, for Actinobacillus actinomycetemcomitans (A.a.) and two other oral bacteria has stimulated this study of the relation between A.a. and periodontal destruction. A.a. has been associated with oral abscesses, brain abscesses, endocarditis and localized juvenile periodontitis. However, in all periodontal studies A.a. also has been observed in 'ordinary' periodontal pockets and subjects without active periodontal destruction. Based on longitudinal studies it is concluded, that there is no proof for a causal relationship between A.a. and periodontal destruction, that the detection of A.a. is not indicative for future alveolar bone loss and that the sensitivity of a test of A.a. is too low for the diagnosis of progressive periodontitis. The sheer presence of A.a. in spite of its virulence factors is not sufficient to cause periodontal destruction. It is stated that, in order to prevent overtreatment and medicalization, the use of bacteriological tests and broad spectrum antibiotics should be restricted to patients with progressive periodontal destruction not responding to standard treatment. PMID- 11908457 TI - [Vestibuloplasty in patients with cleft lip and palate]. AB - After cleft lip repair the upper lip is sometimes attached at the premaxilla. The scar bands and contractures may occur deleterious effects on: the growth, the facial expression, the speech, problems during orthodontic treatment and in prosthetic dental care, regression of the attached gingiva, resorption of the transplanted bone and the aspect of the upper lip. In nine edentulous patients with cleft lip palate, in six patients with cleft lip palate and with a mutilated dentition and in ten younger patients with cleft lip palate the buccal sulcus was successfully restored. The importance of a free upper lip and adequate sulcus has been under-emphasized in treatment of the patient with cleft lip palate. PMID- 11908458 TI - [Quality of care: concepts and definitions]. AB - Quality of dental care is receiving more and more attention. In the past many different definitions of quality are used. The definition of quality as it is used in the Dutch medical care system, is discussed. The policy for quality assurance of the Dutch government and the Dutch Dental Association (NMT) is briefly mentioned. PMID- 11908459 TI - [Turkish and Moroccan migrants in the Netherlands. II. Some background information]. PMID- 11908460 TI - Proceedings of the 2nd Asian-Pacific Symposium on Pain Control. Sydney, Australia, 26-28 January 2001. PMID- 11908461 TI - The interview dance: getting what you ask for. AB - Interviewing potential employees is hardly a science, but what we ask and how we ask it can often elicit the information we need about a prospective employee. Conversely, irrelevant questions can often backfire on us. PMID- 11908462 TI - Five-star service. AB - For the second part in the series of articles featuring human resources initiatives in Michigan hospitals (part one appeared in the November/December issue of Michigan Health & Hospitals magazine), the spotlight is on Lakeland Regional Health System, with three hospitals in southwest Nichigan. It has changed its culture of care to be more oriented toward service. PMID- 11908463 TI - Preparing for the next phase. PMID- 11908464 TI - Potential signalling pathways underlying corticotrophin-releasing hormone mediated neuroprotection from excitotoxicity in rat hippocampus. AB - In several neurological disorders including cerebral ischaemia, glutamate has been implicated as a neurotoxic agent in the mechanisms leading to neuronal cell death. The role of corticotrophin-releasing hormone (CRH), the 41-amino acid peptide, which activates the HPA axis in response to stressful stimuli, remains controversial. In this study, we report that CRH in low physiological concentrations (2 pM), prevented glutamate-induced neurotoxicity via receptor mediated mechanisms when administered to organotypic hippocampal cultures both during and after the glutamate-induced insult. Detailed investigations on the mechanisms mediating this neuroprotective effect showed that activation of the adenylate cyclase pathway and induction of MAP kinase phosphorylation mediate the CRH action. In addition we showed that CRH can inhibit the phosphorylation of JNK/SAPK by glutamate. Most importantly, we showed that CRH can afford neuroprotection against neurotoxicity up to 12 h following the insult, suggesting that CRH is acting at a late stage in the neuronal death cycle, and this might be important in the development of novel neuroprotective agents in order to improve neuronal survival following the insult. PMID- 11908465 TI - Effects of streptozotocin-diabetes on the hippocampal NMDA receptor complex in rats. AB - In animal models of diabetes mellitus, such as the streptozotocin-diabetic rat (STZ-rat), spatial learning impairments develop in parallel with a reduced expression of long-term potentiation (LTP) and enhanced expression of long-term depression (LTD) in the hippocampus. This study examined the time course of the effects of STZ-diabetes and insulin treatment on the hippocampal post-synaptic glutamate N-methyl-D-aspartate (NMDA) receptor complex and other key proteins regulating hippocampal synaptic transmission in the post-synaptic density (PSD) fraction. In addition, the functional properties of the NMDA-receptor complex were examined. One month of STZ-diabetes did not affect the NMDA receptor complex. In contrast, 4 months after induction of diabetes NR2B subunit immunoreactivity, CaMKII and Tyr-dependent phosphorylation of the NR2A/B subunits of the NMDA receptor were reduced and alphaCaMKII autophosphorylation and its association to the NMDA receptor complex were impaired in STZ-rats compared with age-matched controls. Likewise, NMDA currents in hippocampal pyramidal neurones measured by intracellular recording were reduced in STZ-rats. Insulin treatment prevented the reduction in kinase activities, NR2B expression levels, CaMKII-NMDA receptor association and NMDA currents. These findings strengthen the hypothesis that altered post-synaptic glutamatergic transmission is related to deficits in learning and plasticity in this animal model. PMID- 11908467 TI - The effect of tibolone on the lipoprotein profile of postmenopausal women with type III hyperlipoproteinemia. AB - OBJECTIVE: To investigate the short-term effect of treatment with tibolone on plasma lipid and lipoprotein levels in postmenopausal women with type III hyperlipoproteinemia (HLP). DESIGN AND INTERVENTION: Patients were randomized to receive, in a double-blind cross-over fashion, a fixed dose of tibolone, 2.5 mg once daily or placebo for 8 weeks. The two treatment periods were separated by a wash-out period of 6 weeks. At each visit body weight and blood pressure were determined. Before and after each treatment period, fasting venous blood samples were obtained from the patients for biochemical measurements. SETTING: The Leiden University Medical Center. SUBJECTS: Postmenopausal women with type III HLP (aged < or = 65 years) were recruited from the Lipid Clinics of the Leiden University Medical Center, the Amsterdam Medical Center, the Utrecht Medical Center and the University Hospital Rotterdam. Five out of 25 women with type III HLP were eligible to be included in the study. Four of the five included patients completed the study according to the protocol. One patient was excluded from blinded therapy because total cholesterol levels increased above 20 mmol L(-1). MAIN OUTCOME MEASURES: A significant reduction of plasma triglyceride, total cholesterol, VLDL cholesterol and VLDL triglyceride levels. RESULTS: Plasma triglyceride and total cholesterol levels decreased from 6.82 +/- 3.58 to 2.45 +/ 1.36 mmol L(-1) and from 13.53 +/- 3.64 to 6.61 +/- 2.03 mmol L(-1), respectively (both P < 0.05). The body mass index remained unchanged. The glycated haemoglobin percentage decreased significantly from 5.8 to 5.3%. Treatment with tibolone resulted in a profound reduction in plasma apolipoprotein E, VLDL cholesterol and VLDL triglyceride levels (mean reductions of 66, 77 and 70%, respectively, P < 0.05). CONCLUSIONS: Tibolone is a valuable adjuvant to current therapy in postmenopausal women with type III HLP. PMID- 11908466 TI - Photic regulation of L-arginine uptake in the golden hamster retina. AB - One of the limiting steps in the regulation of nitric oxide (NO) synthesis is the availability of its precursor, L-arginine, which depends on the presence of a specific uptake system. A characterization of the L-arginine uptake mechanism in the golden hamster retina was performed. This mechanism was stereospecific, saturable, and monophasic, with an apparent of 56.1 +/- 2.0 microM and a maximum velocity of 36.0 +/- 2.8 pmol/mg prot/min. The basic amino acids L-lysine and L ornithine but not D-arginine or the nitric oxide synthase inhibitors, N(omega) nitro-L-arginine methyl ester and N(omega)-nitro-L-arginine impaired L-arginine influx. Preincubation with L-lysine for 1 h prior to the transport assay significantly stimulated L-arginine uptake. Saturation studies of L-arginine uptake performed at 12.00 and 24.00 h indicated a higher value of Vmax at midnight than at midday. When the hamsters were placed under constant darkness or constant light for 48 h and killed at equivalent time points, representing subjective day and subjective night, the differences in L-arginine influx disappeared. Semiquantitative RT-PCR analysis showed that the levels of mRNAs for both CAT-1 and CAT-2B were significantly higher at midnight than at midday. L Arginine significantly increased cGMP accumulation in a time-dependent manner, with maximal effects during the night. Based on these results, it might be presumed that hamster retinal L-arginine uptake is regulated by the photic stimulus. PMID- 11908468 TI - Sarcoidosis--a great mimicker. PMID- 11908471 TI - Medi-sim multimedia: Clinical Simulations in Pharmacology I. PMID- 11908472 TI - Medication error reduction training. PMID- 11908469 TI - Clinical evaluation of cervical dentin sensitivity (CDS) in patients attending general dental clinics (GDC) and periodontal specialty clinics (PSC). AB - AIM: The objective of this study was to compare the prevalence, severity and distribution of CDS in patients attending general dental clinics (GDC) and periodontal specialty clinics (PSC) and to correlate them to possible causal factors. MATERIAL AND METHODS: 2 groups of patients aged 20-60 years recruited from GDC (144) and PSC (151) were evaluated for CDS by means of a questionnaire and intraoral clinical examinations. Furthermore, gingival recession and plaque scores were recorded at the same visit. RESULTS: The results showed that patients referred to PSC had a significantly higher prevalence of CDS (60.3%) than those examined at GDC (42.4%) (p<0.001). Also, mean plaque scores of PSC patients (1.87 +/- 0.88) was found to be significantly higher than that of GDC (1.44 +/- 0.7) (p<0.01). The occurrence and extent of gingival recession associated with hypersensitive teeth was significantly higher in PSC than GDC patients (p<0.01), with a 5% incidence of severe recession (5 mm) in PSC only. The association of periodontal disease and periodontal treatment to the high prevalence of CDS and gingival recession in PSC patients would suggest their role in predisposition to hypersensitivity. The distribution of CDS in tooth types revealed that upper molars and lower anteriors of PSC patients were mainly affected, and followed by, to a lesser extent, lower right canine and right first molars of GDC patients. CONCLUSION: The prevalence of CDS among our periodontal patients appears somewhat lower than that reported in periodontal specialty clinics of earlier studies but still higher than those reported in other dental populations. This indicates that periodontal disease and its treatments may increase the occurrence of hypersensitivity. PMID- 11908473 TI - Diagnostic ultrasound of fetal anomalies. PMID- 11908470 TI - Clinical and microbiological studies of periodontal disease in Sjogren syndrome patients. AB - BACKGROUND: Little is known about the periodontal status of patients with Sjogren's Syndrome (SS), a chronic inflammatory autoimmune disease characterized by xerophthalmia and xerostomia. The aim of the present study was to evaluate whether the periodontal status of SS patients, in terms of clinical and microbiological parameters, differs from systemically healthy age- and gender matched controls. METHODS: 8 primary SS and 10 secondary SS patients were examined in comparison with 11 control subjects. All patients were diagnosed by the European Community Criteria. Control subjects were systemically healthy and not undergoing periodontal treatment. The comparison of clinical status was made in terms of mean periodontal parameters (plaque index, gingival index, gingival recession, probing pocket depth, probing attachment level and bleeding on probing) as well as the frequency distribution of probing pocket depth and probing attachment level measurements. Microbiological assays of the subgingival dental plaque samples were carried out by both a chairside enzyme test (Periocheck) for the detection of peptidase activity (PA) and a polymerase chain reaction (PCR) analysis for 9 selected periodontal micro-organisms (Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Prevotella intermedia, Treponema denticola, Porphyromonas gingivalis, Eikenella corrodens, Campylobacter rectus, Bacteroides forsythus, Streptococcus oralis). RESULTS: The occurrence, severity and extent of periodontal lesions were not significantly different between the 3 patient groups for all periodontal parameters examined. No significant differences in the sub-gingival plaque samples from control, primary or secondary SS patients for the PA test, frequency or type of periodontal micro-organisms observed. CONCLUSION: No significant differences could be detected in either clinical or microbiological parameters of primary or secondary SS patients compared with that of control subjects. The results of the present study thus support the notion that the periodontal status of patients with SS do not differ from systemically healthy age- and gender-matched controls. PMID- 11908474 TI - Navigating the mental health Internet. PMID- 11908475 TI - NursingStation 2000 (version 2.0). PMID- 11908476 TI - Old hearts, diastolic dysfunction, and blockpnea. PMID- 11908477 TI - Evaluation of a specific balance and coordination programme for individuals with a traumatic brain injury. AB - The purpose of this study was to evaluate the effectiveness of an aerobic dancing training, designed to reduce postural imbalance and coordination deficits for individuals who had sustained a traumatic brain injury (TBI). A two group experimental design was conducted. A control group participated in a traditional muscular training (TMT) programme while participants in the experimental group were assigned to an aerobic dancing, Slide and Step training programme (specific training group (ST)). Participants were evaluated pre- and post-training. Balance was quantified using a force platform and coordination using a Peak Performance system to compare the velocity profiles of a modified Jumping jack test. Results showed that temporal variables were significantly different pre- and post training for the ST group, but no changes were found in the TMT group. The results of the balance test indicated a significant reduction of postural sway area in the ST group but not in the TMT group. Overall, the combination workout with Step and Slide is more effective in reducing balance and coordination deficits when compared to muscular based training. PMID- 11908478 TI - [Characterization of the NPH2 gene, coding for the glomerular protein podocin, implicated in a familial form of cortico-resistant nephrotic syndrome transmitted as an autosomal recessive]. PMID- 11908479 TI - [Renal interstitial fibrosis and urotelial carcinomas after ingestion of a Chinese herb (Aristolochia fangchi)]. PMID- 11908481 TI - American College of Cardiology Clinical Expert Consensus Document on Standards for Acquisition, Measurement and Reporting of Intravascular Ultrasound Studies (IVUS). A report of the American College of Cardiology Task Force on Clinical Expert Consensus Documents developed in collaboration with the European Society of Cardiology endorsed by the Society of Cardiac Angiography and Interventions. PMID- 11908480 TI - [30 years: Happy birthday, GPCR. The bradykinin B2 receptor: an alternative and antiproliferative pathway]. AB - The bradykinin B2 receptor which belongs to the G-protein-coupled receptor (GPCR) family is known for its proliferative effects. The mitogenic signalling pathways used are "classical" pathways for GPCRs. Recent data from our laboratory show that bradykinin can also induce anti-mitogenic effects in proliferating cells using an "alternative" signal transduction pathway involving a protein tyrosine phosphatase. Besides this alternative signalling pathway for the B2 receptor, a novel concept of GPCR signalling is described. PMID- 11908482 TI - Risk analysis for a radio-carpal joint replacement. AB - Risk analysis is required by the medical device directives to provide evidence that manufacturers have eliminated or reduced risks as far as possible so that a medical device does not compromise the safety of patients or health workers. This paper presents a risk analysis for the Swanson wrist implant, which is made from an implantable-grade silicone elastomer and used to replace the radiocarpal joint in the rheumatoid wrist. The main hazards identified were that the implant fractures and that silicone synovitis occurs in patients. The results of this risk analysis will be used to aid the design of a new wrist implant. PMID- 11908483 TI - Investigation of wear of knee prostheses in a new displacement/force-controlled simulator. AB - The performance of two knee simulators designed by ProSim (Manchester, UK) was evaluated by comparison of the wear seen in the press-fit condylar (PFC) Sigma (DePuy) knee prosthesis. Twelve specimens of the same design and manufacturing specification, were subjected to a wear test of 2 x 10(6) cycles duration using bovine serum as a lubricant. The anterior/posterior displacement and internal/external rotation inputs were based on the kinematics of the natural knee. International Standards Organization (ISO) standards were used for the flexion and axial load. The wear rates and wear scar areas were compared across all stations. The mean wear rates found were 17.6+/-5 mm3/10(6) cycles for stations 1 to 6 and 19.6+/-4 mm3/10(6) cycles for stations 7 to 12, resulting in an overall mean wear rate of 18.1+/-3 mm3/10(6) cycles. The differences between the two simulators were not significant. The average wear scar area seen on inserts from stations I to 6 was calculated at 32.4+/-1 per cent of the intended articulating surface. Similarly on stations 7 to 12 the average wear scar area was 30.7+/-3 per cent. The wear scars seen were a good physiological representation of those found from clinical explant data. This study has shown good repeatability from the simulator, both within and between the simulators. PMID- 11908484 TI - A spatial model of the knee for the preoperative planning of knee surgery. AB - A model on the spatial mechanical behaviour of the passive knee is presented. The femoral articular surfaces were represented by generalized, sagittally elliptical, toroidal surfaces. The medial and lateral tibial articular surfaces were represented by a dished spherical surface and the lower hemihyperbolic region of a torus respectively. Anatomical articular cartilage, knee ligaments and the posterior capsule were represented by spring-like deformable elements with non-linear load versus deflection characteristics. All the forces that act on the femur relative to the tibia were represented by three orthogonal forces and three associated moments. Spatial, articulation-dependent femorotibial kinematic constraint equations of the passive knee were formulated in an analytically explicit manner, based on the natural coordinates of the articular surfaces. The constraint equations were solved algebraically in closed form. Equations were derived that describe spatial femoro-tibial motion, ligament length, ligament strain, ligament-based elastic potential energy and the quasi static equilibrium of the passive knee. Software was written, simulations on the motion characteristics and load versus deflection characteristics of the knee were carried out and graphical results were presented. The simulation of planar flexion/extension was almost spontaneous. The time taken to simulate spatial six degree-of-freedom femoro-tibial motion was less than 2.5 min. The models were found to be capable of representing real-life passive knees to a high degree of satisfaction. It has been demonstrated that the models can provide knee surgeons with additional information on major aspects of the preoperative planning of knee surgery. The models can be used to enhance the preoperative planning of ligament reconstruction, articular surfaces related surgery, osteotomy and patellar tendon transfer surgery. PMID- 11908485 TI - Assessment of the ergonomically optimal operating surface height for laparoscopic surgery. AB - PURPOSE: The aim of this study was to find the ergonomically optimal operating surface height for laparoscopic surgery in order to reduce discomfort in the upper extremities of the operators and the assistants. The operating surface height was defined as the level of the abdominal wall of a patient with pneumoperitoneum. MATERIALS AND METHODS: Two pelvi-trainer tests were performed. One test was performed on six different operating surface heights. The (extreme) joint excursions of the shoulder, elbow, and wrist were measured by a video analysis method. Another test was performed by holding a laparoscope for 15 minutes while an electromyelograph of the biceps brachii was made. The results of both tests were evaluated subjectively by a questionnaire. RESULTS: The ergonomically optimal operating surface height lies between a factor 0.7 and 0.8 of the elbow height of the operator/assistant. At this height, the joint excursions stay in the neutral zone for more than 90% of the total manipulation time, and the activity of the biceps brachii when holding the laparoscope stays within 15% of the maximum muscle activity. CONCLUSIONS: The operating surface height influences the (extreme) upper joint excursions of the surgeon. The ergonomically optimal operating surface height reduces the discomfort in the shoulders, back, and wrists of the surgeon during laparoscopic surgery. This optimal table height range for laparoscopic surgery is lower than those currently available. PMID- 11908486 TI - Laparoscopic repair of morgagni's hernia with percutaneous placement of suture. AB - Simultaneous laparoscopic repair of Morgagni's hernia and laparoscopic cholecystectomy in a 59-year-old lady is described. In contrast to most previous reported series, in which repairs were fashioned with direct intracorporeal suturing, stapling, or mesh placement, an alternative technique of suture placement with a percutaneously inserted needle was applied. The procedure was smooth, and there was no evidence of recurrent hernia 9 months after the operation. PMID- 11908487 TI - The effect of chronic flow changes on brachial artery diameter and shear stress in arteriovenous fistulas for hemodialysis. AB - BACKGROUND: Vessel wall adaptation to acute or chronic flow changes is regulated by shear stress (SS) at the endothelium. This hypothesis was tested in the brachial artery (BA) of patients receiving an arteriovenous fistula (AVF) for hemodialysis vascular access. METHODS: The acute and sustained effects were evaluated in 13 patients. Pre-operatively and postoperatively on predetermined time-points BA diameter and shear rate (SR) were measured. SS was calculated from whole blood viscosity and SR. Analysis was performed with Wilcoxon's test and ANCOVA multivariate analysis. RESULTS: Acutely, mean SS increased (475%, p<0.05), peak-to-peak SS decreased (37%, p<0.05) and peak SS remained constant. BA diameter increased (15%, p<0.05). After one year a further increase was observed (r=0.59, p<0.001), plus an increase in mean SS (r=0.78, p<0.001). Peak-to-peak SS remained constant. CONCLUSION: Our results indicate that after AVF placement an acute increase in SS results in an acute increase of vessel diameter. However, one year of sustained high blood flow does not result in restoration of mean SS. PMID- 11908488 TI - A new expandable cannula to increase venous return during peripheral access cardiopulmonary bypass surgery. AB - Peripheral cannulation for cardiopulmonary bypass (CPB) is of prime interest in minimally invasive open heart surgery. As CPB is initiated with percutaneous cannulae, venous drainage is impeded due to smaller vessel and cannula size. A new cannula was developed which can change shape in situ and therefore may improve venous drainage. An in vitro circuit was set-up with a penrose latex tubing placed between the preload reservoir and the cannula, encasing the cannula's inlet and simulating the vena cava. The preload (P) was stabilised at 2 and at 5 mmHg respectively. The maximum flow rate was determined for 4 conditions: passive venous drainage (PVD) and assisted venous drainage (AVD) using a centrifugal pump at the 2 preload settings. We compared the results of the prototype cannula to classical femoral venous cannulae: basket 28Fr, a thoracic 28Fr and a percutaneous 27Fr. Under PVD conditions and a CVP of 2 mmHg, the prototype cannula's flow rate outperformed the next best cannula by 14% (p=0.0002) and 13% under AVD conditions (p=0.0001). Under PVD conditions and a CVP of 5 mmHg, the prototype cannula outperformed the percutaneous cannula by 19% (p=0.0001) and 14% under AVD conditions (p=0.0002). The new cannula outperforms the classical percutaneous venous cannulae during all of the four conditions tested in vitro. PMID- 11908489 TI - Coronary artery bypass grafting in patients with severe left ventricular dysfunction: a prospective randomized study on the timing of perioperative intraaortic balloon pump support. AB - In this prospective trial the results of preoperative and intraoperative IABP in coronary artery bypass graft (CABG) patients with low left ventricular ejection fraction (LVEF) were compared. Sixty CABG patients with preoperative LVEF < or = 0.30 were enrolled: in group A patients (n=30) IABP was started within 2 hours preoperatively; in group B (n=30) it was instituted intraoperatively before weaning from cardiopulmonary bypass. Cardiac performance was assessed through Swan-Ganz catheter monitoring and daily echocardiography. Hospital survival, length of IABP support, intubation, ICU and hospital stay, need for postoperative inotropic drugs and incidence of myocardial infarction were compared between the two groups. Survival in group A patients proved significantly higher (P=0.047). Cardiac performance after myocardial revascularization improved in both groups with significantly better outcomes in group A patients (P<0.001). Doses of inotropic drugs (dobutamine, enoximone) were lower in group A (P=0.001; P=0.004) and duration shorter (P<0.001; P<0.001). No major IABP-related complication was observed. PMID- 11908490 TI - Donor age and mechanosensitivity of human bone cells. AB - With increasing age the human skeleton decreases in density, thereby compromising its load-bearing capacity. Mechanical loading activates bone formation, but an age-dependent decrease in skeletal mechanoresponsiveness has been described in rats. In this paper we examine whether age-related bone loss is reflected by a decrease in the mechanosensitivity of isolated bone cells from human donors. Bone cell cultures were obtained from 39 donors (males and females) between 7 and 85 years of age. Cultures were challenged with 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) or mechanically stressed by treatment with pulsating fluid flow (PFF; 0.7 +/- 0.03 Pa at 5 Hz for 1 h). The growth capacity of the bone-derived cell population almost halved between 7 and 85 years of age. Basal alkaline phosphatase activity of the cells increased with donor age, while the response to 1,25(OH)2D3, measured as stimulated osteocalcin production, decreased with age. Together this suggests that the cell cultures from older donors represented a more mature, slower-growing cell population than the cultures from young donors. All cell cultures responded to mechanical stress with enhanced release of prostaglandin E2 (PGE2) and I2 (PGI2). The magnitude of the response was positively correlated with donor age, cell cultures from older donors showing a higher response than cultures from younger donors. There was also a positive correlation between time to reach confluency and mechanosensitivity, i.e., the PGE2 response to PFF treatment was higher in bone cell cultures with a slower growth rate. We conclude that bone cell cultures from older donors have a lower proliferative capacity and a higher degree of osteoblastic maturation than younger donors. The higher degree of osteoblastic maturation explains the higher response of the cultures to mechanical stress, in line with earlier studies on chicken bone cells. This study found no evidence for loss of mechanosensitivity with donor age. The reduced growth capacity might, however, be a factor in age related bone loss. PMID- 11908491 TI - A randomized trial of sodium fluoride (60 mg) +/- estrogen in postmenopausal osteoporotic vertebral fractures: increased vertebral fractures and peripheral bone loss with sodium fluoride; concurrent estrogen prevents peripheral loss, but not vertebral fractures. AB - Postmenopausal Caucasian women aged less than 80 years (n = 99) with one or more atraumatic vertebral fracture and no hip fractures, were treated by cyclical administration of enteric coated sodium fluoride (NaF) or no NaF for 27 months, with precautions to prevent excessive stimulation of bone turnover. In the first study 65 women, unexposed to estrogen (-E study), age 70.8 +/- 0.8 years (mean +/ SEM) were all treated with calcium (Ca) 1.0-1.2 g daily and ergocalciferol (D) 0.25 mg per 25 kg once weekly and were randomly assigned to cyclical NaF (6 months on, 3 months off, initial dose 60 mg/day; group F CaD, n = 34) or no NaF (group CaD, n = 31). In the second study 34 patients, age 65.5 +/- 1.2 years, on hormone replacement therapy (E) at baseline, had this standardized, and were all treated with Ca and D and similarly randomized (FE CaD, n = 17; E CaD, n = 17) (+E study). The patients were stratified according to E status and subsequently assigned randomly to +/- NaF. Seventy-five patients completed the trial. Both groups treated with NaF showed an increase in lumbar spinal density (by DXA) above baseline by 27 months: FE CaD + 16.2% and F CaD +9.3% (both p = 0.0001). In neither group CaD nor E CaD did lumbar spinal density increase. Peripheral bone loss occurred at most sites in the F CaD group at 27 months: tibia/fibula shaft 7.3% (p = 0.005); femoral shaft -7.1% (p = 0.004); distal forearm -4.0% (p=0.004); total hip -4.1% (p = 0.003); and femoral neck -3.5% (p = 0.006). No significant loss occurred in group FE CaD. Differences between the two NaF groups were greatest at the total hip at 27 months but were not significant [p < 0.05; in view of the multiple bone mineral density (BMD) sites, an alpha of 0.01 was employed to denote significance in BMD changes throughout this paper]. Using Cox's proportional hazards model, in the -E study there were significantly more patients with first fresh vertebral fractures in those treated with NaF than in those not so treated (RR = 24.2, p = 0.008, 95% CI 2.3-255). Patients developing first fresh fractures in the first 9 months were markedly different between groups: -23% of F CaD, 0 of CaD, 29% of FE CaD and 0 of E CaD. The incidence of incomplete (stress) fractures was similar in the two NaF-treated groups. Complete nonvertebral fractures did not occur in the two +E groups; there were no differences between groups F CaD and CaD. Baseline BMD (spine and femoral neck) was related to incident vertebral fractures in the control groups (no NaF), but not in the two NaF groups. Our results and a literature review indicate that fluoride salts, if used, should be at low dosage, with pretreatment and co treatment with a bone resorption inhibitor. PMID- 11908492 TI - Effects of transdermal estradiol delivered by a matrix patch on bone density in hysterectomized, postmenopausal women: a 2-year placebo-controlled trial. AB - This 2-year, double-masked, randomized, placebo-controlled trial was designed to evaluate the safety and efficacy in preventing bone loss in postmenopausal women of two doses of transdermal 17betaestradiol (estradiol) delivered by a matrix patch, compared with placebo. One hundred and sixty healthy, hysterectomized postmenopausal volunteers aged 40-60 years with serum estradiol levels < 20 pg/ml were started on treatment at four centers in The Netherlands. Every 6 months, bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DXA) at the lumbar spine, non-dominant wrist and left hip, and markers of bone turnover were assessed in urine and serum. The treatment arms were: estradiol, 100 microg/day (E-100, n = 53), oestradiol, 50 microg/day (E-50, n = 54), placebo (P-100, placebo to E-100, n = 27 or P-50, placebo to E-50, n = 26). Treatment was continued for up to 2 years. After 24 months, BMD of the lumbar spine in the E 100 group differed by 7.7% [5.8-9.5%] (mean [95% confidence interval]) from the placebo group and showed a mean (s.e.m.) increase in BMD from baseline of 5.9% (0.69%). For the E-50 group the difference compared with placebo was 6.2% [4.4 8.0%] and the absolute increase was 4.5% (0.62%); in the placebo group, the absolute change was -2.3% (0.48%). In the total wrist, the changes were: E-100: difference compared with placebo 2.5% [1.5 3.6%], absolute increase 0.6% (0.3%); E-50: difference compared with placebo 2.9% [1.8-3.9%], absolute increase 0.7% (0.25%); and absolute change on placebo: -2.5% (0.35%). In the total hip, the changes were: E-100: difference compared with placebo 3.7% [2.2-5.2%], absolute increase 2.8% (0.5%); E-50: difference compared with placebo: 3.2% [1,8-4.7%], absolute change 2.4% (0.36%); and absolute change on placebo -1.4% (0.66%). Three markers of bone turnover--serum bone-specific alkaline phosphatase, serum osteocalcin and urinary CTX--fell significantly during the trial. Breast pain was reported by 8% of women on placebo, by 6% of women on E-50 and by 17% of women on E-100. Estradiol delivered by the E-50 matrix patch effectively reversed bone loss in hysterectomized postmenopausal women with few side-effects. The marginal additional gain in BMD with the higher dose may be offset by a more important side effect profile. PMID- 11908493 TI - Thalidomide in Behcet's disease. AB - Behcet's disease is a chronic relapsing multisystem disease of unknown aetiology. It has a relapsing cyclical course, and is characterized by the triad of aphthous stomatitis, genital ulcerations and uveitis. There is familial and geographical clustering of cases, especially around the Mediterranean, the Middle East and East Asia. The condition is uncommon, but lesions sometimes are recalcitrant and can be debilitating to the affected individual. The treatment of Behcet's currently involves the use of steroids, immunomodulaters and immunosuppressives. Thalidomide has been used in cases of Behcet's disease with some success. This review will discuss Behcet's disease and the current information we have about using thalidomide for its treatment. PMID- 11908494 TI - Single molecular detection of a perylene dye dispersed in a Langmuir-Blodgett fatty acid monolayer using surface-enhanced resonance Raman scattering. AB - The Langmuir-Blodgett (LB) monolayer technique was used to fabricate single molecule LB monolayer containing bis(phenethylimido)perylene (PhPTCD), a red dye dispersed in arachidic acid (AA) with an average doping of 1 molecule per microm2. The monolayer was transferred onto Ag island films to obtain spatially resolved surface-enhanced resonance Raman scattering (SERRS) spectra. The mixed LB monolayers were fabricated with a concentration, on average, of 1, 6, 19 and 118 PhPTCD molecules per microm2 in AA. The AA provides a two-dimensional host matrix whose background signal does not interfere with the detection of the probe molecule's SERRS signal. The properties of the single molecule detection were investigated using micro-Raman with a 514.5-nm laser line. The Ag island surfaces coated with the LB monolayer were mapped with spatial steps of 3 microm and global chemical imaging of the most intense SERRS band in the spectrum was also recorded. The SERRS and surface-enhanced fluorescence (SEF) of the neat and single molecule LB monolayer were recorded in a temperature range from liquid nitrogen to + 200 degrees C. Neat PhPTCD LB monolayer spectra served as reference for the identification of characteristic signatures of the single molecule behavior. The spatial resolution of Raman-microscopy experiments, the multiplicative effect of resonance Raman and SERRS, and the high sensitivity of the new dispersive Raman instruments, allow SERRS to be part of the family of single molecular spectroscopies. PMID- 11908495 TI - Semiglobal simplex optimization and its application to determining the preferred solvation sites of proteins. AB - The classical simplex method is extended into the Semiglobal Simplex (SGS) algorithm. Although SGS does not guarantee finding the global minimum, it affords a much more thorough exploration of the local minima than any traditional minimization method. The basic idea of SGS is to perform a local minimization in each step of the simplex algorithm, and thus, similarly to the Convex Global Underestimator (CGU) method, the search is carried out on a surface spanned by local minima. The SGS and CGU methods are compared by minimizing a set of test functions of increasing complexity, each with a known global minimum and many local minima. Although CGU delivers substantially better success rates in simple problems, the two methods become comparable as the complexity of the problems increases. Because SGS is generally faster than CGU, it is the method of choice for solving optimization problems in which function evaluation is computationally inexpensive and the search region is large. The extreme simplicity of the method is also a factor. The SGS method is applied here to the problem of finding the most preferred (i.e., minimum free energy) solvation sites on a streptavidin monomer. It is shown that the SGS method locates the same lowest free energy positions as an exhaustive multistart Simplex search of the protein surface, with less than one-tenth the number of minizations. The combination of the two methods, i.e.. multistart simplex and SGS, provides a reliable procedure for predicting all potential solvation sites of a protein. PMID- 11908497 TI - Quantum mechanical study of the conformational behavior of proline and 4R hydroxyproline dipeptide analogues in vacuum and in aqueous solution. AB - The conformational behavior of the title compounds has been investigated by Hartree-Fock, MP2, and DFT computations on the most significant structures related to variations of the backbone dihedral angles, cis/trans isomerism around the peptide bond, and diastereoisomeric puckering of the pyrrolidine ring. In vacuum the reversed gamma turn (gammal), characterized by an intramolecular hydrogen bridge, corresponds to the absolute energy minimum for both puckerings (up and down) of the pyrrolidine ring. An additional energy minimum is found in the helix region, but only for an up puckering of the pyrrolidine ring. When solvent effects are included by means of the polarizable continuum model the conformer observed experimentally in condensed phases becomes the absolute minimum. The down puckering is always favored over its up counterpart, albeit by different amounts (0.4-0.5 kcal/mol for helical structures and about 2 kcal/mol for gammal structures). In helical structures cis arrangements of the peptide bond are only slightly less stable than their trans counterparts. This is no longer true for gammal structures, because the formation of an intramolecular hydrogen bond is possible only for trans peptide bonds. In most cases, proline and hydroxyproline show the same general trends; however, the electronegative 4(R) substituent of hydroxyproline leads to a strong preference for up puckerings irrespective of the backbone conformation. PMID- 11908496 TI - Semiautomatic sequence-specific assignment of proteins based on the tertiary structure--the program st2nmr. AB - The sequence-specific assignment of resonances is still the most time-consuming procedure that is necessary as the first step in high-resolution NMR studies of proteins. In many cases a reliable three-dimensional (3D) structure of the protein is available, for example, from X-ray spectroscopy or homology modeling. Here we introduce the st2nmr program that uses the 3D structure and Nuclear Overhauser Effect spectroscopy (NOESY) peak list(s) to evaluate and optimize trial sequence-specific assignments of spin systems derived from correlation spectra to residues of the protein. A distance-dependent target function that scores trial assignments based on the presence of expected NOESY crosspeaks is optimized in a Monte Carlo fashion. The performance of the program st2nmr is tested on real NMR data of an alpha-helical (cytochrome c) and beta-sheet (lipocalin) protein using homology models and/or X-ray structures; it succeeded in completely reproducing the correct sequence-specific assignments in most cases using 2D and/or 15N/13C Nuclear Overhauser Effect (NOE) data. Additionally to amino acid residues the program can also handle ligands that are bound to the protein, such as heme, and can be used as a complementary tool to fully automated assignment procedures. PMID- 11908498 TI - Restrained point-charge models for disaccharides. AB - Various methods for deriving atomic partial charges from the quantum chemical electrostatic potential and moments have been tested for the sucrose molecule. We show that if no further information is used, the charges on some carbon atoms become large and charge patterns involving these atoms are badly determined and poorly transferable. Adding lone-pairs on the ether oxygen atoms or dividing the molecule into smaller fragments did not cure the instabilities. We develop a method, CHELP-BOW0, that restrains charges toward zero with different weights for different atoms. These harmonic restraints preserve the linear form of the least squares equations, which are solved in a single step using singular-value decomposition. CHELP-BOW0 improves the chemical transferability of the charges compared to unrestrained methods, and slightly improves their conformational transferability. It introduces a modest degradation of the fit compared to unrestrained CHELP-BOW (mean average deviation of the potential 0.00016 vs. 0.00010 a.u.). A second new method, CHELP-BOWC, avoids the need for restraints by including several conformations in the fit, weighting each according to its estimated energy in solution. CHELP-BOWC charges are more transferable than CHELP BOW or CHELP-BOW0 charges to conformations not included in the training set. Restraints to zero charge do not further improve transferability of the CHELP BOWC charges. We, therefore, recommend CHELP-BOW charges for rigid molecules and CHELP-BOWC charges for flexible molecules. PMID- 11908499 TI - On the irrelevance of electrostatics for the crystal structures and polymorphism of long even n-alkanes. AB - It is known that the experimental triclinic crystal structures of even n-alkanes are not well reproduced upon energy minimization with current force fields. The inclusion of electrostatics does not solve this, and, moreover, some charge schemes show unphysical features such as positively charged carbon atoms or charge alternation. The effect of the electrostatics on the energies of the crystal structures of the even n-alkanes, and thereby on their polymorphism, has never been established. A new charge scheme is introduced that yields physically sensible charges without constraints. It will also be shown, however, that electrostatics are relevant neither for the structures of the crystals, nor for their energies. PMID- 11908500 TI - Oligovalent link atoms in embedding calculations. AB - Making reasonable statements about large systems is a notoriously difficult task of quantum chemistry. If such a system consists of a small but electronically important region, the core, which requires a treatment on a high level of theory, and a larger remainder, the bulk, which only acts in a perturbative fashion and thus admits a description on a lower level of theory, then it is appropriate to partition it accordingly and use an embedding scheme to provide for the coupling of the two regions. In many cases of said partitioning it will be necessary to cut covalent bonds so that the emerging free valences have to be saturated by link atoms. Mostly hydrogen atoms are used. However, it may be necessary to separate the entire system so that bulk atoms are generated that are bound to two or three core atoms at once, that is, such bulk atoms are divalent or even trivalent with respect to the core. In the present article we demonstrate by means of several silane molecules, ranging from Si3H8 to Si104H92, that the use of oligovalent link atoms is quite promising. PMID- 11908501 TI - Accompanying coordinate expansion formulas derived with the solid harmonic gradient. AB - A series of accompanying coordinate expansion (ACE) formulas for calculating the electron repulsion integral (ERI) over both generally and segmentally contracted solid harmonic (SH) Gaussian-type orbitals (GTOs) can be rederived by the use of the modified operator (called solid harmonic gradient here) of the spherical tensor gradient of Bayman and the reducing solid harmonic gradient defined in this article. The final general formulas contain the reducing mixed solid harmonics defined in a previous article [Ishida, K. J Chem Phys 1999, 111, 4913] and the reducing triply mixed solid harmonics defined previously [Ishida, K. J Chem Phys 2000, 113, 7818]. Each general formula in the series is named ACEb1k1, ACEb2k3, or ACEb3k3. New general algorithm can be obtained inductively from the general formula named ACEb2k3, in addition to the previously developed ACEb1k1 and ACEb3k3. For calculating ERI practically, we select one of these ACE algorithms, as it gives the minimum floating-point operation (FLOP) count. Theoretical assessment by the use of the FLOP count is performed for the (LL/LL) class of ERIs over both generally and segmentally contracted SH-GTOs (L = 1-3). It is found that the present ACE is theoretically the fastest among all rigorous methods in the literature. PMID- 11908502 TI - Theoretical Auger electron spectra of polymers by density functional theory calculations using model dimers. AB - We propose a new approach for analysis of Auger electron spectra (AES) of polymers by density functional theory (DFT) calculations with the Slater's transition-state concept. Simulated AES and X-ray photoelectron spectra (XPS) of four polymers [(CH2CH2)n (PE), (CH2CH(CH3))n (PP), (CH2CH(OCH3))n (PVME), and (CH2CH(COCH3))n (PVMK)] by DFT calculations using model dimers are in a good accordance with the experimental ones. The experimental AES of the polymers can be classified in each range of 1s-2p2p, 1s-2s2p, and 1s-2s2s transitions for C KVV and O KVV spectra, and in individual contributions of the functional groups from the theoretical analysis. PMID- 11908503 TI - Dynamics of a protein and water molecules surrounding the protein: hydrogen bonding between vibrating water molecules and a fluctuating protein. AB - Internal and rigid-body motions of bovine pancreatic trypsin inhibitor (BPTI) and of water molecules surrounding the BPTI are studied in a vicinity of an energy minimum using a normal mode analysis proposed as the independent molecule model. Water's rigid-body motion is predominant in comparison to its internal motions. We have derived information about the relationship between the magnitude of a thermal ellipsoid of an H-bonding atom and the anisotropy of its ellipsoid, and the relationship between the magnitude of the ellipsoid and the H-bond strength. We see a relationship between vibrational frequencies (assuming rigid-body motion of the water molecules) and the H-bond strength of the water taking part in this H-bonding. Analyzing the H-bond strength, we found that a hydrogen in water is likely to H-bond to oxygen in the protein, whereas an oxygen in water has a less strong preference to H-bond to the protein. For water molecules acting as the hydrogen acceptor, strong H-bonding has longer lifetimes than weak H-bonding. PMID- 11908504 TI - Theoretical study on the mechanism of the gas-phase reaction of diborane(3) anion with carbon disulfide. AB - The complex potential energy surface of the gas-phase reaction of HB(H)BH- with CS2 to give three low-lying products [B2H3S]- + CS, [BH2CS]- + HBS, and [BH3CS] + BS-, involving nine [B2H3CS2]- isomers and 12 transition states, has been investigated at the CCSD(T)/6-311++G(d,p)/B3LYP/6-311++G(d,p) level. Our calculations are in harmony with the recent experimental and theoretical results, and reveal some new bonding and kinetic features of this reaction system. Our theoretical results may help the further identification of the products [BH2CS]- + HBS and [BH3CS] + BS- and may provide useful information on the chemical behaviors of other electron-deficient boron hydride anions. PMID- 11908505 TI - Software news and updates. Basis-set completeness profiles in two dimensions. AB - A two-electron basis-set completeness profile is proposed by analogy with the one electron profile introduced by D. P. Chong (Can J Chem 1995, 73, 79). It is defined as Y(alpha, beta) = sigmam sigman (Galpha(1)Gbeta(2)/(1/r12)/ psim(1)psin(2)) (psim(1)psin(2)/r12/Galpha(1)Gp(2)) and motivated by the expression for the basis-set truncation correction that occurs in the framework of explicitly correlated methods (Galpha is a scanning Gaussian-type orbital of exponent alpha and [psim] is the orthonormalized one-electron basis under study). The two-electron basis-set profiles provide a visual assessment of the suitability of basis sets to describe electron-correlation effects. Furthermore, they provide the opportunity to assess the quality of the basis set as a whole- not only of the individual angular momentum subspaces, as is the case for the one electron basis-set profiles. The two-electron completeness profiles of the cc pVXZ (X = D, T, Q), aug-cc-pVTZ, cc-pCVTZ, and SVP-auxiliary basis sets for the carbon atom are presented as illustrative examples. PMID- 11908506 TI - CQ Sources/Bibliography. Meanings and contexts: anthropological perspectives in bioethics. PMID- 11908507 TI - Cancer and the prothrombotic state. AB - Thrombosis is a frequent complication of cancer, so it follows that the presence of a tumour confers a prothrombotic state. Indeed, in patients with cancer, each of the three components of Virchow's triad that predispose for thrombus formation have abnormalities, thus fulfilling the requirement for a prothrombotic or hypercoagulable state. The many signs and symptoms of the prothrombotic state in cancer range from asymptomatic basic abnormal coagulation tests to massive clinical thromboembolism, when the patient may be gravely ill. Many procoagulant factors, such as tissue factor and cancer procoagulant, are secreted by or are expressed at the cell surface of many tumours. Platelet turnover and activity are also increased. Damaged endothelium and abnormalities of blood flow in cancer also seem to play a part, as does abnormal tumour angiogenesis. Some studies have even suggested that these abnormalities may be related to long-term prognosis and treatment. We briefly describe the various clinical manifestations of thrombosis in cancer and discuss the evidence for the existence of a prothrombotic or hypercoagulable state associated with this disease. Further work is needed to examine the mechanisms leading to the prothrombotic state in cancer, the potential prognostic and treatment implications, and the possible value of quantifying indices of hypercoagulability in clinical practice. PMID- 11908508 TI - Cancer and ethnic minorities. PMID- 11908509 TI - Penetrating vascular trauma in Johannesburg, South Africa. AB - An awareness that time crucially affects outcome underpins the principles of management of vascular injury. Patients with hard signs of vascular injury should undergo urgent exploration. Soft signs mandate investigation, and arteriography is still the standard of care. Noninvasive vascular imaging may prove its worth in the future. All patients with penetrating arterial injury should receive broad spectrum antibiotic prophylaxis. Early repair of carotid artery injury provides the best likelihood of a neurologically intact survivor. There is a definite and emerging role of endovascular therapy both for difficult access injuries and for the later complications of vascular injury, such as false aneurysm and arteriovenous fistulas. The experimental and clinical evidence for the use of intraluminal shunts in peripheral vascular injury is compelling, and experience in their use is accumulating. Vascular trauma is complex and ideally is carried out by experts in a multidisciplinary environment; resuscitation and prompt revascularization are likely to lead to satisfactory outcomes. The major trauma load in South Africa represents an unparalleled experience in management of vascular injury, which seems likely to continue for the foreseeable future. PMID- 11908510 TI - Why do hypoxic cells behave badly? PMID- 11908511 TI - Is the increase in bronchial responsiveness or FEV1 shortly after cessation of beta2-agonists reflecting a real deterioration of the disease in allergic asthmatic patients? A comparison between short-acting and long-acting beta2 agonists. AB - Regular use of beta2-agonists might result in increased bronchial hyper responsiveness (BHR) and decreased forced expiratory volume in 1 sec (FEV1). It has been suggested that these possible detrimental effects are not a real deterioration of the disease, but that it might be only a transient (rebound) effect shortly after discontinuing this regular use. Moreover, these effects are thought to occur especially during short-acting and not during long-acting beta2 agonists use. The aim of this study was to invest gate whether a rebound effect (a pharmacological deterioration effect diminishing after several hours) in FEV1 and PC20 (concentration of histamine causing a 20% fall in FEV1 with regard to baseline) occurred after cessation of regular use of beta2-agonists, and whether this occurred both after short-acting and long-acting beta2-agonists. Allergic asthmatic patients (n = 134) were randomly allocated to the use of a short-acting (salbutamol), a long-acting beta2-agonist (formoterol) or placebo for 12 weeks (double-blind, double-dummy). No other asthma medication was allowed, including inhaled corticosteroids. At the start and every 4 weeks later FEV and PC20 were measured, each time at least 12 h after the last doses of study medication, which is in the possible rebound period. To investigate whether a (transient) rebound effect occurred, parameters were additionally measured at least 72 h later after discontinuation of the study medication. After 12 weeks of short-acting beta2 agonist use, a drop was seen in FEV1 from 85.6 (+/- 2.21)% predicted to 78.8 (+/- 2.9)% predicted, measured 15 h (median) after the last doses of medication. This was significantly different compared to placebo. When measured 168 h (median) later FEV1 recovered to 85.5 (+/- 2.4)% predicted, comparable to baseline. PC20 decreased with -1.17 (+/- 0.44) doubling dose after 12 weeks of short-acting beta2-agonist use, measured 15 h after the last doses of medication, which was significantly different compared to placebo. However, 168 h later PC20 recovered slightly with +0.55 (+/- 0.34) doubling dose, but this value was still lower compared to placebo. In contrast, during long-acting beta2-agonist and placebo use no significant changes were seen. In conclusion, the use of short-acting beta2-agonists resulted in a transient (rebound) effect in FEV while the effects on PC20 may point to a real deterioration of the disease. Long-acting beta2 agonist and placebo use showed no changes. We conclude that a mono-therapy of short-acting and not of long-acting beta2-agonists might have deleterious effects in asthma. PMID- 11908512 TI - Plasminogen activator inhibitor-I (PAI-I) polymorphisms in patients with obstructive sleep apnoea. AB - Cardiovascular diseases are frequent among patients with the obstructive sleep apnoea syndrome (OSAS), The aetiopathogenesis of this association is unclear. Type 1 plasminogen activator inhibitor (PAI-1) is one of the primary regulators of the fibrinolytic system. A reported association between PAI-1 activity and an insertion/deletion polymorphism (4G/5G) in the promoter region of the PAI-1 gene suggests a critical role for this genomic region in the pathogenesis of several cardiovascular diseases. In this study, we determined the prevalence of this polymorphism in patients with OSAS and in healthy control subjects. The 4G/5G polymorphism in the promoter region of the PAI-1 gene was determined in 78 male patients with severe OSAS (56 +/- 2 apnoeas per hour) and in 70 healthy male, non smoker volunteers of similar age, without personal or familial history of cardiovascular disease. The frequency ofthe 4G/4G, 4G/5G and 5G/5G genotypes in patients with OSAS (18%, 62%, 19%, respectively) was not significantly different from that seen in healthy subjects (16%, 60%, 24% P=NS). These results show that the distribution of the 4G/5G polymorphism in the promoter region ofthe PAI-1 gene in patients with OSAS is similar to that observed in healthy subjects. This observation suggests that the PAI-1 polymorphism has no relationship with the increased risk of cardiovascular diseases seen in patients with OSAS. PMID- 11908513 TI - An efficient and reproducible method for measuring hydrogen peroxide in exhaled breath condensate. AB - We investigated the sensitivity and reproducibility of a test procedure for measuring hydrogen peroxide (H202) in exhaled breath condensate and the effect of storage of the condensate on the H2O2 concentration, and compared the results to previous studies. Twenty stable COPD patients breathed into our collecting device twice for a period of 10 min. The total exhaled air volume (EAV) and condensate volume were measured both times and the H2O2 concentration of the condensate was determined fluorimetrically. The concentration was measured again after freezing the reaction product at -70 degrees C for a period of 10, 20 and 40 days. We collected 2-5 ml condensate in 10 min. The EAV and condensate volumes were strongly correlated. There was no significant difference between the mean H2O2 concentration of the first and second test. We obtained a detect on limit for the H2O2 concentration of 0.02 micromoll(-1). The H2O2 concentration appeared to remain stable for a period up to 40 days of freezing. Compared to previous studies we developed a more efficient breath condensate collecting device and obtained a lower H2O2 detection limit. The measurement of exhaled H2O2 was reproducible. In addition, storage of the samples up to 40 days showed no changes in H2O2 concentration. PMID- 11908514 TI - The enemy within: keeping self-reactive T cells at bay in the periphery. AB - The remarkable capacity of the mammalian immune system to coordinate deadly attacks against numerous invading pathogens, yet turn a blind eye to self-tissues continues to fascinate immunologists. It has been clear for some time that immune cells capable of recognizing self-proteins exist in normal individuals without seemingly causing harm. The 'peripheral tolerance' mechanisms that keep these cells in check are the focus of intense research, not least because defects in these pathways might cause autoimmune diseases. In this review, new developments in our understanding of peripheral tolerance are discussed. PMID- 11908515 TI - Population exposure to diagnostic use of ionizing radiation in The Netherlands. AB - The use of ionizing radiation for diagnostic medical procedures and the exposure of the Dutch population to this radiation were assessed for 1998. The annual average effective dose from diagnostic medical exposures has increased by 26% to 0.59 mSv per capita since the last inventory of medical radiation exposure in the Netherlands a decade ago. The population-averaged effective dose comprises x-ray procedures in hospitals (87%), nuclear medicine examinations (11%), mammography screening (1.5%), and extramural dentistry (0.2%). The rise has resulted mainly from an increase in frequency and patient dose for CT examinations and from vascular radiology. The increase in the number of CT examinations leveled off in the mid-1990's. Medically exposed people were found to be significantly older than the general population. Based on age distribution alone, an "age reduction factor" for the risk coefficient of 0.64 was found to apply to the medically exposed group. More information on patient dose for the complete set of procedures should, according to this study, become available. PMID- 11908516 TI - An examination of uranium levels in Canadian forces personnel who served in the Gulf War and Kosovo. AB - A uranium bioassay program was conducted involving 103 active and retired Canadian Forces personnel. The total uranium concentrations in each of two 24-h urine collections were analyzed separately at independent commercial laboratories by inductively coupled plasma mass spectrometry (ICP-MS) and by instrumental neutron activation analysis (INAA). The mean and median concentrations were determined to be 4.5 ng L(-1) and 2.8 ng L(-1), respectively, from ICP-MS and 17 ng L(-1) and 15 ng L(-1), respectively, from INAA. The total uranium concentrations were sufficiently low so that isotopic (238U:235U ratio) assays could not be performed directly from urine samples. Isotopic assays were performed on hair samples from 19 of the veterans participating in the testing. The isotopic hair assays were scattered around the natural 238U:235U ratio of 137.8, ranging from 122 +/- 21 to 145 +/- 16 (1sigma). Due to concern expressed in the media over possible depleted uranium exposure and long-term retention in bone, a single bone sample (vertebrate bone marrow) from a deceased member of the Canadian Forces was also analyzed for total uranium content and isotopic ratio by ICP-MS. The sample was shown to have 16.0 +/- 0.3 microg kg(-1) uranium by dry weight and a 238U:238U isotopic ratio of 138 +/- 4, consistent with natural uranium. PMID- 11908518 TI - The unique instability of Irish demography. PMID- 11908519 TI - The history of tracheotomy. AB - The history of tracheotomy, from the documentation in ancient Egyptian times through to the formalisation of the open technique by Chevalier Jackson in the twentieth century, is outlined. PMID- 11908517 TI - Developmental regulation and downstream effects of the knox class homeobox genes Oskn2 and Oskn3 from rice. AB - Plant homeobox genes of the class 1 knox (knotted1-like) type are involved in the regulation of shoot apical meristem formation and function. Their expression generally occurs either throughout the meristem or specifically at the lateral organ boundaries. Down-regulation in the organ primordia is tightly controlled and misexpression in leaves leads to a perturbed development. Here, the transcriptional control of two rice knox genes, Oskn2 and Oskn3, was addressed, showing that the promoter sequences of both genes mediate the initial down regulation during lateral organ formation, but are insufficient to keep expression in lateral organs stably off. Therefore, maintenance of the repressed state requires control elements outside the promoter regions. Ectopic expression of Oskn2 or Oskn3 induced similar defects in panicle branching. internode elongation and leaf patterning. However, small differences suggested that their target gene specificities are not completely overlapping. This was supported by the observation that Oskn3 protein but not Oskn2 could interact with two reported recognition sequences of a KNOX protein from barley. Finally, protein-protein interactions may contribute to the functioning of KNOX proteins, as the ability of Oskn3 and Oskn2 to form heterodimers could be demonstrated. PMID- 11908520 TI - Rudolf Virchow (1821-1902) and Die Cellularpathologie (1858). AB - Rudolf Virchow gave clinical significance to Schwann's cell theory and thereby changed the course of pathology. Although he intended to found a science of pathological physiology based on omnis cellula e cellula, advances in microscopy facilitated and promoted, even dictated, the development of histopathology. As well as Die Cellularpathologie and Die krankhaften Geschwulste, Virchow has left the Archiv which he edited until shortly before his death in 1902. In later life he turned increasingly to anthropology, archaeology, and ethnology, but from an early age he was renowned as a fiery liberal politician. PMID- 11908521 TI - Life support system using water electrolysis. AB - The purpose of the integrated physiologico-hygienic and biotechnological laboratory experiment was to evaluate the effect of life support subsytems placed inside a sealed cabin on the cabin environment and, consequently, on the human performance and activity, and also to assess the operability of subsystems by the crewmen and disclose any resulting difficulties. PMID- 11908522 TI - [Grmek's library donation to IMEC]. PMID- 11908524 TI - [The Tuesday seminar]. AB - Grmek as a member of Jouanna's group of scholars on Tuesday morning at the Sorbonne. PMID- 11908523 TI - [Grmek, medical history, and paleopathology]. AB - Mirko Drazen Grmek died on 6 March 2000, defeated by an implacable enemy (amyotrophic lateral sclerosis, which had been diagnosed just 18 months earlier). He has now found peace in his final resting place, the Montparnasse cemetery in Paris. His immense body of work reveals an omniscient man of great wisdom, a cosmopolitan polyglot who devoted his life to the history of science, with particular emphasis on medicine and disease. He looked at paleopathology for what that discipline could bring to the study of populations in antiquity, and succeeded in anchoring it in history with his definitive concept of "pathocenose", created in 1969. Several years later, his most important work, "Les Maladies a l'aube de la civilisation occidentale", (1983) set forth with definitive and convincing illustrations the importance of paleopathology, which will therefore be forever associated with one of the most outstanding medical history books of the XXth. century. PMID- 11908525 TI - Life sciences and space research XII. Proceedings of the Open Meeting of the Working Group on Space Biology of the Sixteenth Plenary Meeting of COSPAR, Constance, F.R.G., 23 May-5 June 1973. PMID- 11908526 TI - Interaction between radiation effects, gravity and other environmental factors in Tribolium confusum. AB - Multicellular organisms possess homeostatic control systems, which, in responding to changes of the external environment, modify the internal milieu of the organism accordingly, in order to make survival and normal physiological processes possible. This group has studied effects on physiological processes of single and combined environmental factors in the flour beetle, Tribolium confusum. Studies included low- and high-LET radiation, gravity compensation, near-weightlessness in space flight, ambient temperature, atmospheric composition, and magnetic field effects on the growth and development of Tribolium. For somatic effects, there appears to be a "normal physiological range" for each of these environmental variables; moreover, their effects seem independent of each other. When one of the listed environmental factors is near the limits of normal physiological tolerance, however, marked synergism has been observed between the effects of this factor and other environmental stresses. Fertilized Tribolium eggs showed a differential radiosensitivity to external irradiation, and a linear, dose-effect relationship was obtained, as accelerated heavy nitrogen and oxygen ions were used. Synergism was found when Tribolium were irradiated with ionizing radiation and subsequently exposed to temperature either below or above their normal physiological range. Gravity compensation or near weightlessness in space flight have adverse effects on the development of Tribolium when an ambient temperature is near the higher or lower limit of tolerance. Similar results were observed when pupae were exposed to a combination of oxygen, temperature and magnetic field stresses. PMID- 11908527 TI - Retinal change induced in the primate (Macaca mulatta) by oxygen nuclei radiation. AB - Retinas of primates (Macaca mulatta) were exposed to oxygen nuclei at the Bevatron, Berkeley, California. Color fundus photographs and fluorescein angiograms were taken of the retinas prior to irradiation and up to 5 weeks post exposure. Animals were sacrificed at post exposure intervals for histopathologic examination of the retinas. A series of animals were exposed to 200 kVp X-ray and examined on the same regime as the first series. The results showed a low rad equivalent dose for retinal damage as compared with the X-ray series, i.e., a high quality factor, and a marked compression of the latency between exposure and onset of the retinal pathology. PMID- 11908528 TI - The Biostack Experiments I and II aboard Apollo 16 and 17. AB - The concept of the Biostack experiment has become practicable through European scientific collaboration and with help of NASA. The objectives of this experiment flown aboard Apollo 16 and 17 are to study the biological effects of individual heavy cosmic particles of high-energy loss (HZE) not available on earth; to study the influence of additional spaceflight factors; to get some knowledge on the mechanism by which HZE particles damage biological materials; to get information on the spectrum of charge and energy of the cosmic ions in the spacecraft; to estimate the radiation hazards for man in space. For this purpose the Biostack experiment comprises a widespread spectrum of biological objects, and various radiobiological end-points are under investigation. Bacterial spores, protozoa cysts, plant seeds, shrimp eggs, and insect eggs were included in the Biostack experiment packages together with different physical radiation detectors (nuclear emulsions, plastics, AgCl crystals, and LiF thermoluminescence dosimeters). By using special arrangements of biological objects and physical track detectors, individual evaluation of tracks was obtained allowing the identification of each penetrating particle in relation to the possible biological effects on its path. The response of the different biological objects to space flight and HZE ions bombardment was of different degree, presumably depending on the ability of the organism to replace the cells damaged by a hit. The results help to estimate the radiation hazard for astronauts during space missions of long duration. PMID- 11908529 TI - Experimental methods of correlation between the trajectories of cosmic heavy ions and biological objects: dosimetric results from the Biostack experiment on Apollo 16 and 17. AB - The biological objects in the Biostack are stacked alternately between physical detectors of HZE particle tracks which include nuclear emulsions. The pattern of the biological objects is transferred to the upper side of the adjacent emulsion by optical illumination. On each sheet of nuclear emulsion a coordinate grid is transferred to the bottom side by optical illumination. The visible track left by the passage of the heavy ion can then be very clearly localized. The charge of the ions is determined by photometric measurements of the tracks. These measurements are calibrated with known tracks of heavy ions accelerated at the Bevatron at Berkeley. The results are given as: flux, number of stars per cm3, charge of the ions, comparison with other detectors (plastics, AgCl crystals, LiF), energy loss, hit region in the biological objects. PMID- 11908530 TI - AgCl detectors in the Biostack II experiment aboard Apollo 17. AB - Two layers of AgCl detectors with a total surface of 90 cm2 were flown. Tracks of nuclei, from light (Z>4) up to the heaviest were recorded and could be distinguished by their geometrical trackwidths. The tracks were divided into five groups of atomic numbers, and their abundance was measured. Also the number of surviving nuclear stars was counted. 22.5 cm2 of the detector surface were covered with eggs of Artemia salina. The detectors could be developed without removing the eggs, so that the spots hit could be determined directly. The radiation effect on these eggs is being investigated. PMID- 11908531 TI - Personal growth and personality development: a foreword to the special section. PMID- 11908532 TI - Defense mechanisms, behavior, and affect in young adulthood. AB - The relationship between defense mechanism use, observed behavior, and affect was investigated in a sample of 91 young adults. Defense mechanisms were assessed using Cramer's (1991a) Defense Mechanism Manual for TAT stories; behavior was based on observer Q-sort ratings (Block, 1978). The findings show that men and women who rely on the immature defense of denial at age 23 show multiple signs of behavioral immaturity, as well as anxiety. In contrast, extensive use of projection was related to a suspicious, hyperalert personality style, including anxiety and depression, in men, but to a sociable, nonwary, nondepressed style in women. The use of the mature defense of identification, by women, was related to behavior characterized by maturity, social competence, and the absence of depressive symptoms. PMID- 11908533 TI - Emotional responses to changing feedback: is it better to have won and lost than never to have won at all? AB - Two experiments investigated how self-esteem guides people's emotional responses to changing evaluative feedback. In both experiments, participants received an initial evaluation (either positive or negative) followed by a second evaluation (either positive or negative). Emotional reactions to the second evaluation were then assessed. High self-esteem participants found feedback that was consistently negative to be most distressing, whereas low self-esteem participants were most disturbed by feedback that changed from positive to negative. The theoretical and practical implications of the findings are discussed. PMID- 11908534 TI - Being known, intimate, and valued: global self-verification and dyadic adjustment in couples and roommates. AB - We contend that close relationships provide adults with optimal opportunities for personal growth when relationship partners provide accurate, honest feedback. Accordingly, it was predicted that young adults would experience the relationship quality with relationship partners who evaluated them in a manner consistent their own self-evaluations. Three empirical tests of this self-verification hypothesis as applied to close dyads were conducted. In Study 1, young adults in dating relationships were most intimate with and somewhat more committed to partners when they perceived that partners evaluated them as they evaluated themselves. Self-verification effects were pronounced for those involved in more serious dating relationships. In Study 2, men reported the greatest esteem for same-sex roommates who evaluated them in a self-verifying manner. Results from Study 2 were replicated and extended to both male and female roommate dyads in Study 3. Further, self-verification effects were most pronounced for young adults with high emotional empathy. Results suggest that self-verification theory is useful for understanding dyadic adjustment across a variety of relational contexts in young adulthood. Implications of self-verification processes for adult personal development are outlined within an identity negotiation framework. PMID- 11908535 TI - Personal goals and psychological growth: testing an intervention to enhance goal attainment and personality integration. AB - We hypothesized that semester goal attainment provides a route to short-term psychological growth. In an attempt to enhance this process, we randomly assigned participants to either a goal-training program or to a control condition. Although there were no main effects of program participation on later goal attainment, important interactions were found. Consistent with a "prepared to benefit" model, participants already high in goal-based measures of personality integration perceived the program as most useful and benefited the most from the program in terms of goal attainment. As a result, they became even more integrated and also increased in their levels of psychosocial well-being and vitality. Implications for theories of short-term growth and positive change are discussed, as is the unanswered question of how to help less-integrated persons grow. PMID- 11908536 TI - The Michelangelo phenomenon and personal well-being. AB - Previous research has demonstrated that when a close romantic partner views you and behaves toward you in a manner that is congruent with your ideal self, you experience movement toward your ideal self (termed the "Michelangelo phenomenon"; Drigotas, Rusbult, Wieselquist, & Whitton, 1999). The present research represents an attempt demonstrate the phenomenon's link to personal well-being. Results of a cross-sectional study of individuals in dating relationships, with a 2-month follow-up assessing breakup, replicated previous findings regarding relationship well-being and revealed strong links between the model and personal well-being, even when accounting for level of relationship satisfaction. Such results provide further evidence for the social construction of the self and personal well-being. PMID- 11908537 TI - Personality development and growth in women across 30 years: three perspectives. AB - This article addresses three questions about personality development in a 30-year longitudinal study of women (N = 78): (1) To what extent did the women maintain the same position in relation to each otheron personality characteristics over the 30 years, and what broad factors were related to the amount of change in their rank order? (2) Did the sample as a whole increase or decrease over time on indices of personality growth, and did they change in ways distinctive to women? (3) Were experiential factors associated with individual differences in the amount of change? Results showed that personality was quite consistent while also showing that time interval was positively related to rank-order change and age was negatively related to rank-order change. Over the period from age 21 to age 52, the women increased on measures of norm-orientation and complexity and showed changes on measures of Dominance and Femininity/Masculinity consistent with the hypothesis that changing sex roles would lead to increases in Dominance and increases, then decreases, in Femininity/Masculinity. A third set of results showed that changes in Dominance and Femininity/Masculinity were associated with life circumstances such as marital tension, divorce, and participation in the paid labor force. The implications of the findings for personality development and growth are discussed. PMID- 11908538 TI - Impact of organizational restructuring on nurses' facilitation of parental participatory care. AB - In a study to explore nurses' perceptions of their role as facilitators of parent participation in the care of hospitalized children and the factors that promoted or impeded their role, massive reform and downsizing of hospitals were noted to have created tension between the restructured health care system and the capacity of nurses to provide the optimal level of care they valued. Specifically, revised staffing patterns, (e.g., increased number of part-time and casual nurses) and workload issues (e.g., time constraints, early discharges) created concerns relative to job satisfaction and the quality of participatory care facilitated by nurses. Findings from this study support those reported in other literature. PMID- 11908539 TI - The effect of staff nurse participation in a clinical nursing research project on attitude towards, access to, support of and use of research in the acute care setting. AB - The purpose of this study was to determine the effect of participation in research on staff nurses' attitude towards, access to, perceived support of and reported use of research in practice. Six medical surgical units in a southeastern Ontario teaching hospital were randomly assigned to receive 3 different levels of exposure to research: high, low and usual. On the high participation units, a clinical research group consisting of the investigator and interested nurses (n = 18) critiqued research literature related to an important clinical issue (i.e., patterns of sleep) and designed and implemented a clinical research study. On the low participation units, a similar clinical research group (n = 10) met once and were involved, solely, in the design and implementation of the clinical research protocol. On the control units, there were no formalized research groups or activities. All registered nurses (n = 235), including the research group participants, on the 6 units were surveyed with a research utilization questionnaire (RUQ) pre and post participant intervention. The RUQ scores were higher on the high participation units at baseline and post intervention in comparison to the low and control units. Nurses who participated directly in the clinical research groups (high and low) reported similar RUQ scores post intervention and higher scores in comparison to all nurses. All RUQ scores were higher post intervention. Nurses with clinical expertise but minimal research expertise participated meaningfully in clinical research. While participation had an individual effect there was no unit effect, suggesting other factors, such as organizational support and culture, are important determinants of research use. PMID- 11908540 TI - Increasing nurse first-line leader (manager) positions to improve nurse retention and quality of patient care. AB - Nurses' job action during the spring and summer 2001 taught us much in British Columbia. Post job action de-briefing with non-contract staff confirmed for the Capital Health Region in Victoria that the workload of patient care managers, along with their scope of responsibility was contributing to dissatisfaction, both on the part of patient care managers as well as nursing staff who they are intended to serve. Further, it was becoming increasingly apparent that the success of most of the organizational initiatives depended greatly on the presence, availability and support of patient care managers. Supported by the senior leadership team, the Chief Nursing Officer for the Capital Health Region agreed to have a look at unit based leadership--see what was being discussed across the country and in the literature and draw on her own experience in three provinces. The result would be a discussion paper that would circulate broadly throughout the organization and would provide the organization with broad input to help it decide collectively "What do we need to do? Where do we need to go from here? And how will we get there?" This article is an adaptation of that discussion paper. PMID- 11908541 TI - Elizabeth Logan: demonstrating a nursing model in practice. PMID- 11908542 TI - "Let's get real". Choosing silence--choosing voice. PMID- 11908543 TI - Supply and demand for cardiac nurses in Ontario: perceptions of CNOs. AB - This article presents the results of a nursing survey of cardiac care hospitals undertaken by a Cardiac Care Network of Ontario Consensus Panel on Cardiovascular Human Resources. The focus of the Panel was to identify areas of current or pending shortages in human resources and make recommendations to the Ministry of Health and Long-Term Care about human resource management in adult cardiac care in Ontario. The article presents the number and mix of full-time, part-time and casual nursing staff, the age distribution of RNs, and the number of vacant Registered Nurse (RN) positions for a sample of cardiac care hospitals in Ontario. Next a sample of Chief Nursing Officer opinions about factors contributing to current difficulties in recruiting RNs and the outlook for future shortages are presented. Implications for nurse managers are offered, including development of new recruitment and retention strategies, identification of further efficiencies in care provision, and a need for nurse manager involvement in debates about the future of how health care is provided in Canada. PMID- 11908544 TI - Effects of SCH23390 and raclopride on a run-climb-run behavioral task in rats. AB - The present study was designed to compare the putative differential behavioral consequences of treatment with SCH23390 (a selective dopamine D1 receptor blocker) and raclopride (a selective dopamine D2 receptor blocker) by employing a run-climb-run (RCR) behavioral task of different lengths. Rats were trained to traverse an uncovered floor alleyway (150 cm), climb a vertical rope (70 or 130 cm), and run across an upper board (100 cm) to access water for the reinforcement. At doses of 0.05, 0.10 and 0.15 mg/kg administered intraperitoneally 60 min before the behavioral session, both SCH23390 and raclopride significantly increased the total time to complete the tasks in a dose related fashion. Microstructural analysis on the RCR behavioral performance revealed that the most apparent impairment induced by either drug was observed as the subject shifted motion from the end of the floor alleyway to the rope when hopping or to initiate climbing. However, the motion shift from climbing to running on the upper board was significantly impaired by raclopride, but not by SCH23390. Surprisingly, neither SCH23390 nor raclopride affected the climbing response itself. Running responses on the floor alleyway board were significantly disrupted by raclopride, whereas those on the upper board were significantly disrupted by SCH23390. Deficits induced by both drugs were more profound for the longer compared to the shorter rope, and were most notably shown at the transition area from running to climbing. These data indicate that both dopamine D1 and D2 receptors are involved in the RCR behavior performance. The results also suggest that the cost of motoric demand for behavioral performance is important for evaluating of the effects of drugs blocking dopamine receptors. PMID- 11908546 TI - The relationship between FTL and NA, DMV or CVLM in central cardiovascular control. AB - The aim of the present study was to examine the relationship between the lateral tegmental field (FTL), a cardioinhibitory area, with other cardioinhibitory areas, i.e., the ambiguus nucleus (NA) and the dorsal motor nucleus of vagus (DMV) and the caudal ventrolateral medulla (CVLM), a vasopressor inhibitory area. In 55 cats anesthetized with chloralose (40 mg/kg) and urethane (400 mg/kg), the cardiovascular responses of heart rate (HR), systemic arterial blood pressure (SAP) and vertebral nerve activity (VNA) were recorded. The FTL, NA, DMV and CVLM were identified first by stimulation (rectangular pulses in 80 Hz, 0.5 ms, 50-100 microA) and then confirmed by microinjection of sodium glutamate (Glu, 0.25M, 70 nl). In studying the influence of NA, DMV, or CVLM lesion on the Gluinduced responses in FTL, kainic acid (KA, 24 mM, 100 nl) was microinjected into the NA, DMV or CVLM. FTL stimulation produced an average decrease of HR by 55%. After KA lesioning of the ipsilateral NA or the DMV, the decreased HR induced by FTL was significantly diminished. After subsequent lesion of the contralateral DMV or NA, the bradycardia of FTL was abolished. The reduction of resting HR was more intense after lesioning the NA than DMV and with the left side more than that of the right side. These studies suggest that the cardioinhibitory responses of FTL are mediated through both NA and DMV with predominance of the former, while the hypotensive effect of FTL is mediated through CVLM. The precise pathway responsible for the FTL-induced bradycardia and hypotension is to be determined. PMID- 11908545 TI - Functional coupling of voltage-dependent L-type Ca2+ current to Ca2+-activated K+ current in pituitary GH3 cells. AB - Ca2+-activated K+ currents (I(K(Ca)) can contribute to action potential repolarization and after-hyperpolarization in GH3 cells. In this study, we examined how the activation of I(K(Ca) at the cellular level could be functionally coupled to Ca2+ influx through L-type Ca2+ channels. A 30-msec Ca2+ influx step to 0 mV was found to exhibit substantial contribution of Ca2+ influx through the activation of I(Ca,L) to the activation of I(K(Ca)). A bell-shaped relationship between the conditioning potentials and the integrated I(K(Ca)) was observed, suggesting that the magnitude of integrated I(Ca,L) correlates well with that of integrated I(K(Ca)) in the same cell. A linear relationship of integrated I(Ca,L) and integrated I(K(Ca)) was found with a coupling ratio of 69+/-7. The value of the coupling ratio was unaffected by the presence of Bay K 8644 or nimodipine, although these compounds could effectively affect the amplitudes of both I(K(Ca)) and I(Ca,L). However, tetrandrine could decrease the coupling ratio. Paxilline or intracellular Ca2+ buffer with EGTA decreased the coupling ratio, while apamin had no effect on it. Interestingly, phorbol 12 myristate 13-acetate also reduced the coupling ratio significantly, whereas thapsigargin increased this value. Thus, the present study indicates that the activation of I(K(Ca)) during brief Ca2+ influx, which is inhibited by paxilline, is coupled to Ca2+ influx primarily through the L-type channels. The selective modulation of I(K(Ca)) by second messengers or Ca2+ release from internal stores may affect the coupling efficiency and hence cellular excitability. PMID- 11908547 TI - The effect of monaural middle ear destruction on postnatal development of mouse inferior colliculus. AB - This study examined the effect of monaural middle ear destruction on postnatal development of inferior collicular (IC) neurons of the laboratory mouse, Mus musculus. Monaural middle ear destruction was performed on juvenile mice and the density, number and size of IC neurons were determined at different postoperative ages. For electrophysiological study, collicular auditory response properties were always examined four weeks after operation. Monaural middle ear destruction produced larger neurons in the ipsilateral IC (relative to the operated ear) and smaller neurons in the contralateral IC of experimental mice in comparison with IC neurons of control mice. IC neurons of control mice typically had lower minimum thresholds and greater Q10 values than IC neurons of experimental mice. In experimental mice, neurons in the contralateral IC typically had longer latencies and higher minimum thresholds than neurons in the ipsilateral IC. Clear tonotopic organization was only observed for IC neurons of control mice. Possible mechanisms for these different observations are discussed. PMID- 11908548 TI - NMDA inhibits oxotremorine-induced acid secretion via the NO-dependent cyclic GMP system in rat stomach. AB - The mechanism of N-methyl-D-aspartate (NMDA) inhibits oxotremorine-induced acid secretion was examined in rat stomach, in relation to the cyclic GMP system. NMDA (10(-7) M) did not affect the spontaneous acid secretion from the everted preparations of isolated rat stomach, but inhibited the acid secretion stimulated by oxotremorine, and this effect of NMDA was antagonized by 2-amino-5 phosphonovaleric acid (AP-5), (+/-)3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) or N(G)-nitro-L-arginine (L-NNA). NMDA also elevated the cyclic GMP content of mucosal slices from rat stomach, and this effect of NMDA was antagonized by L-NNA. These results indicate that NMDA receptors are present in the rat stomach and regulate the gastric acid secretion. The mechanism underlying the effect of NMDA inhibits oxotremorine-induced acid secretion may be mediated by the NO-dependent cyclic GMP system. PMID- 11908549 TI - Role of histamine in aggravation of gastric acid back-diffusion and vascular permeability in septic rats. AB - The aim of the present study was to investigate the role of histamine in aggravation of gastric acid back-diffusion and vascular permeability in lipopolysaccharide (LPS)-induced septic rats. Male specific pyrogen-free Wistar rats were deprived food for 24 h before the experiment. Intravenous LPS (3 mg/kg dissolved in sterilized saline) was given to rats 12 h after food removal. Control rats received sterilized saline only. Under diethylether-anesthesia, the pylorus and esophageal sphincters of rats were ligated. Vagotomy also was performed. The stomachs were then irrigated for 3 h with physiological acid solutions containing 0-150 mM HCl plus adequate amount of NaCl. Increases in various ulcerogenic parameters, such as gastric acid back-diffusion, mucosal histamine concentration, luminal hemoglobin (Hb) content and stomach ulcer, were dependent on the concentration of acid solutions irrigated in stomachs of those LPS rats. Gastric vascular permeability also was increased in an acid concentration-related manner. In those LPS rats, high correlation was found between extents of acid back-diffusion and mucosal ulceration. Increased vascular permeability also closely related to the luminal Hb content. Moreover, these ulcerogenic parameters were dose-dependently ameliorated by intraperitoneal ketotifen and ranitidine. Diamine oxidase also was effective in inhibition, but exogenous histamine on the contrary, produced exacerbation of these ulcerogenic parameters. In conclusion, histamine plays a pivotal role in modulating gastric acid back-diffusion and vascular permeability that are greatly associated with hemorrhagic ulcer in septic rats. PMID- 11908550 TI - Options in disability determination. Lessons that pertain to the regional musculoskeletal disorders. PMID- 11908551 TI - What does cartilage calcification tell us about osteoarthritis? PMID- 11908552 TI - Dynamic T cell receptor clonotype changes in synovial tissue of patients with early rheumatoid arthritis: effects of treatment with cyclosporin A (Neoral). AB - OBJECTIVE: To study T cell receptor (TCR) repertoire changes in synovial membrane over a 16 week period in patients with early rheumatoid arthritis (RA); and to study the influence of cyclosporin A (CSA) on TCR repertoire in a subgroup of these patients. METHODS: Synovial tissue biopsies and paired blood samples were obtained from 12 patients with early RA at 2 time points. Seven patients were treated with CSA (Neoral-Sandimmun, 3 mg/kg/day) and 5 patients with placebo for 16 weeks. TCR V gene repertoires were analyzed by semiquantitative PCR-ELISA. CDR3 spectratyping and sequence analysis was used to compare TCR clonotype distributions. RESULTS: TCR-specific mRNA was detected in all synovial tissue biopsies at the first sampling, but in only 8/12 biopsies 16 weeks later (4/7 CSA group, 4/5 placebo group). Overrepresented TCR BV genes were found in biopsies of 10/12 patients at the first time point, and in 7/12 patients after 16 weeks (3/7 CSA, 4/5 placebo). CDR3 sequence analysis revealed dynamic repertoire changes with only a few persisting clonotypes in the synovial tissue of placebo controls. Persisting T cell clonotypes were more frequently found in the synovial tissue of CSA treated patients compared to the placebo group. CONCLUSION: These data suggest a dynamic process of T cell recruitment in the joints of RA patients. This process, possibly due to activation and subsequent infiltration of new T cell clones, apparently is influenced by CSA treatment. Synovial tissue T cells were no longer detected after 16 weeks' CSA treatment in 3 patients. In the other CSA treated patients, new T cell clones infiltrated, while other clones were persistently represented in the joints. These data may have important consequences for the design of T cell targeted therapies for RA. PMID- 11908553 TI - Norepinephrine from synovial tyrosine hydroxylase positive cells is a strong indicator of synovial inflammation in rheumatoid arthritis. AB - OBJECTIVE: Density of sympathetic nerve fibers in synovial tissue was lower in patients with rheumatoid arthritis (RA) compared to those with osteoarthritis (OA). This was accompanied by norepinephrine (NE) release from synovial tyrosine hydroxylase positive cells (TH+ cells). We investigated the role of TH+ cells and NE in synovial inflammation. METHODS: Synovial tissue of 34 patients with RA and 36 with OA who underwent knee joint replacement surgery was characterized using immunohistochemistry and a synovial tissue superfusion technique, respectively. In culture experiments with mixed synoviocytes, the effect of NE on secretion of interleukin 6 (IL-6), IL-8, tumor necrosis factor (TNF), and matrix metalloproteinase-3 (MMP-3) was investigated. RESULTS: Tissue density of TH+ cells was higher in RA compared to OA (63.9 vs 34.2 cells/mm2; p = 0.017). Basal NE release from synovial tissue correlated highly significantly with density of TH+ cells in RA (Rrank = 0.573, p = 0.001) but not in OA (Rrank = 0.102, NS). Basal NE release correlated with the degree of inflammation in RA (Rrank = 0.420, p = 0.021) but not in OA (Rrank = 0.174, NS), and with spontaneous IL-8 secretion in RA (Rrank = 0.581, p = 0.001) but not in OA (Rrank = 0.160, NS). Only in RA, density of TH+ cells correlated positively with spontaneous secretion of IL-6, IL 8, and MMP-3. We confirmed the extensive loss of sympathetic nerve fibers in RA compared to OA (0.32 vs 3.1 nerve fiber/mm2; p < 0.001). The ratio of sympathetic to sensory nerve fibers was 1 to 5 in RA and 2 to 1 in OA. A ratio of 1.0 separates almost all patients into 2 diseases groups (RA vs OA). Prior prednisolone treatment of RA patients was related to decreased spontaneous cytokine secretion, a lower density of T cells, CD163+ macrophages and TH+ cells, a lower degree of inflammation, and reduced synovial NE secretion. NE was able to inhibit secretion of IL-6 (in OA), IL-8 (in RA), and TNF (in RA and OA) in culture experiments. CONCLUSION: TH+ cells and release of NE are strongly linked to a higher degree of synovial inflammation. Culture experiments indicate that NE has antiinflammatory properties at higher concentrations (10(-5) M). NE secretion of TH+ cells may be an antiinflammatory mechanism to counteract local inflammation. Thus, TH+ cell derived NE can be an important local factor of immunomodulation in synovial inflammation. PMID- 11908554 TI - Dose response and safety study of meloxicam up to 22.5 mg daily in rheumatoid arthritis: a 12 week multicenter, double blind, dose response study versus placebo and diclofenac. AB - OBJECTIVE: This Phase III, placebo and active controlled, multicenter trial evaluated the efficacy and safety of meloxicam 7.5, 15, and 22.5 mg daily for the treatment of rheumatoid arthritis (RA). METHODS: A 12 week, randomized, double blind, double dummy, parallel group trial compared daily oral meloxicam 7.5, 15, and 22.5 mg to placebo (negative control) and diclofenac 75 mg BID (positive control). A total of 894 patients (18 years of age with confirmed RA who flared following an NSAID-free period) were randomized to be treated. Baseline scores for all endpoints were similar among the treatment groups. Patient assessments were at 0, 2, 4, 8, and 12 weeks or early termination. RESULTS: All treatment groups demonstrated significant improvement from baseline (p < 0.001). Meloxicam 7.5 and 22.5 mg was significantly superior to placebo in all 5 primary efficacy endpoints (swollen joint count, tender joint count, patient pain, patient and physician global; all p < 0.05). Diclofenac 150 mg was superior to placebo for 4 of 5 primary efficacy measures (all but swollen joint count; p < 0.05) and meloxicam 15 mg was superior for 3 of 5 primary endpoints (patient pain and patient and physician global). AUC of patient global, patient pain, and modified Health Assessment Questionnaire demonstrated dose-response (p < 0.04), while AUC ACR20 showed a qualitative trend in the same direction. The rate of gastrointestinal (GI) events during the 12 week trial for all doses of meloxicam and diclofenac did not differ significantly from placebo (23.2-32.0%). GI withdrawals were comparable and not significantly different across all treatment groups (4.3-5.7%). CONCLUSION: This trial demonstrated a dose response relationship for meloxicam 7.5, 15, and 22.5 mg using AUC measurement of response for the treatment of RA. All 3 doses of meloxicam. and positive control, were effective in the treatment of RA. The overall incidence rate of GI events did not differ significantly from placebo in either the meloxicam treatment groups or the positive control. PMID- 11908555 TI - A randomized, placebo controlled trial of an antisense oligodeoxynucleotide to intercellular adhesion molecule-1 in the treatment of severe rheumatoid arthritis. AB - OBJECTIVE: To determine the safety of an antisense oligodeoxynucleotide to intercellular adhesion molecule-1 (ICAM-1) (ISIS 2302), administered in an intensive 4 week regimen with dose escalation; and to provide preliminary evidence for efficacy in rheumatoid arthritis (RA). METHODS: Patients with active RA were enrolled in a 6 month, double blind, placebo controlled, dual center, dose escalation (0.5, 1, and 2 mg/kg) study. Subjects received a total of 13 intravenous ISIS 2302 infusions, given on alternate days for 2 weeks and then 3 times a week for another 2 weeks. Doses of corticosteroids (< or = 10 mg/day) and disease modifying antirheumatic drugs (stable > or = 3 months) remained constant throughout the study. The primary efficacy endpoint was the Day 26 Paulus index, with secondary evaluations at Months 2-6. RESULTS: A total of 43 patients were enrolled with 11, 10, 3, and 19 patients receiving placebo or 0.5, 1, or 2 mg/kg of ISIS 2302, respectively. There were no differences between groups after randomization and the mean baseline swollen joint count was 22.5. Pharmacokinetic studies revealed a T(1/2) of 63 min and first-order kinetics with slight dose dependency, suggesting a saturable clearance process, although no accumulation was noted with repeat dosing. The Paulus 20% responses at Day 26 were 20%, 0%, and 5% for patients treated with ISIS 2302 (0.5, 1, 2 mg/kg, respectively) and 36% with placebo. For Months 2-6, the average intent-to-treat Paulus 20% responses were 21.2% for ISIS 2302 and 12.6% for placebo. Only ISIS 2302 treated subjects (19%) achieved Paulus 50% responses. ISIS 2302 was well tolerated. An expected and transient mean activated partial thromboplastin time increase of roughly 7 s was observed at the highest dose (2 mg/kg), as were small and clinically insignificant increases in serum C3a levels. T/B cell immunophenotyping, recall antigen skin testing, and serum immunoglobulin levels revealed no significant immunosuppressive effects. CONCLUSION: This study shows that 13 ISIS 2302 infusions over 4 weeks are well tolerated in patients with active RA. Although significant efficacy was not evident at the primary endpoint (1 month), the study lacked sufficient power to draw any formal conclusions. We tested a 4-fold drug concentration range, which led to a lower area under the curve range than was therapeutic in a subsequent Crohn's disease trial. Any further evaluation of this well tolerated ICAM-1 antisense agent should therefore be conducted at higher dosing. PMID- 11908556 TI - High levels of interleukin 13 in rheumatoid arthritis sera are modulated by tumor necrosis factor antagonist therapy: association with dendritic cell growth activity. AB - OBJECTIVE: To investigate the physiology of interleukin 13 (IL-13) in rheumatoid arthritis (RA) and the effects of tumor necrosis factor (TNF) antagonists (etanercept) on the distribution of IL-13 in patients with RA. METHODS: We measured cytokine levels in RA sera (pre/post etanercept), RA synovial fluid (SF), osteoarthritis (OA) SF, and normal human sera by ELISA. Detection of IL-13 was not influenced by rheumatoid factor, as revealed in spike recovery and isotype antibody control studies. Biologically active IL-13 in RA SF was studied using dendritic cell (DC) progenitors that develop into mature DC with IL-13 and with neutralizing antibodies to IL-13. The modulation of IL-13 by etanercept was compared to that of IL-6 and monocyte colony stimulating factor (M-CSF). The effect of etanercept on the ability of RA sera to promote DC growth was studied using DC progenitors. RESULTS: IL-13 was increased in RA sera versus normal sera, OA SF, and RA SF. Relative to OA SF and normal sera, RA SF was enriched in IL-13. The IL-13 contained in RA samples was biologically active, prompting DC growth from progenitors. Circulating DC growth activity was strongly reduced by anti-TNF therapy. Whereas decreases in DC growth factors including IL-13 and IL-6 occurred with etanercept therapy and were associated with clinical improvement, concurrent increases in circulating M-CSF (a non-DC, monocyte-specific growth factor) were noted. CONCLUSION: The increase of biologically active IL-13 in RA supports the concept that IL-13 regulates immune cell (including dendritic cell) activity and indicates how the varied anatomical distribution of cytokines may play a role in the RA disease process. The differential regulation of circulating IL-13 and M CSF levels by TNF antagonists further implies discrete roles in the TNF-cytokine network in RA. PMID- 11908557 TI - The acute phase response does not fully predict the presence of insulin resistance and dyslipidemia in inflammatory arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA) is associated with an increased mortality rate from cardiovascular disease. This may relate to insulin resistance and dyslipidemia, which were both reported to correlate with the acute phase response in RA. We investigated whether insulin resistance and dyslipidemia could be explained by the acute phase response as well as excess weight in inflammatory arthritis. METHODS: We investigated 87 patients, 38 with RA, 29 with spondyloarthropathy, 20 with undifferentiated inflammatory arthritis. Thirty age, sex, and race matched healthy volunteers served as controls. Fasting blood samples were taken for determination of erythrocyte sedimentation rate (ESR), plasma glucose, serum insulin, and total cholesterol (chol), low density lipoprotein cholesterol (LDL-chol), high density lipoprotein cholesterol (HDL chol), and triglycerides. Insulin resistance was estimated by the homeostasis model assessment for insulin resistance (HOMA) and the quantitative insulin sensitivity check index (QUICKI). RESULTS: In controls the mean (SD) HOMA (microU x mmol/ml x l), QUICKI, body mass index (BMI, kg/m2), and ESR (mm/h) were 1.1 (0.5), 0.393 (0.048), 22.9 (2.8), and 13 (8) in patients; they were 1.9 (1.3), 0.357 (0.037), 26.5 (4.2), and 26 (18) in controls, respectively. Each of these differences was highly significant (p < 0.001). HDL-chol concentrations were lower (p = 0.002) and chol/HDL-chol ratios and triglyceride levels were higher (p < 0.001 and p = 0.004, respectively) in patients compared to controls. A high ESR predicted insulin resistance and dyslipidemia, while a high BMI similarly predicted insulin resistance but not dyslipidemia. After controlling for ESR and BMI, insulin sensitivity was no longer different between patients and controls, while HDL-chol concentrations remained lower (p = 0.015) and chol/HDL-chol ratios remained higher (p = 0.003) in patients compared to controls. CONCLUSION: Insulin resistance and dyslipidemia were highly prevalent in patients with inflammatory arthritis. The acute phase response and excess weight could fully explain the insulin resistance but only partially explain the dyslipidemia. These findings have important implications for the management of inflammatory arthritis. PMID- 11908558 TI - Gastroprotective therapy and risk of gastrointestinal ulcers: risk reduction by COX-2 therapy. AB - OBJECTIVE: Proton pump inhibitors (PPI) and misoprostol decrease the risk of development of nonsteroidal antiinflammatory drug induced gastric ulcers and aid healing of upper gastrointestinal (GI) ulcers. H2 receptor antagonists (H2RA) are less effective for this task, but are widely used by patients and physicians for the treatment of GI symptoms and duodenal ulcers. Sucralfate is a weaker agent that is sometimes used for prophylaxis or treatment of upper GI ulcers. We investigated the effect of GI drugs and selective and nonselective NSAID on the incidence of GI ulcer development in a cohort of patients immediately after the release of celecoxib and rofecoxib to investigate the effect of confounding by indication when effective GI agents and cyclooxygenase 2 (COX-2)-specific inhibitors are prescribed to a high risk population. METHODS: During a 6 month period of observation 8547 NSAID users were evaluated by mailed questionnaire concerning NSAID drug use and ulcer development. In the first half of 1999, patients took 12,177 separate NSAID courses. GI therapy that followed the development of upper GI ulcers was excluded from analysis. Ulcer reports were confirmed by followup validation. RESULTS: GI drugs were used concomitantly in this population by 42% of patients using an NSAID. GI drugs were associated with an increased risk of ulcer. But this risk was confined to PPI (OR 4.1, 95% CI 2.95, 5.69), and not to other GI drugs. Overall, patients using nonselective NSAID compared to those taking COX-2-specific inhibitors had an increased risk of upper GI ulcers (OR 2.12, 95% CI 1.43, 3.34). Patients taking nonselective NSAID plus PPI were also at increased risk for upper GI ulcers compared to those taking nonselective NSAID alone (OR 5.09. 95% CI 3.88, 6.67). Similarly, the risk of upper GI ulcers was increased in the nonselective NSAID plus PPI group (OR 3.83, 95% CI 2.32, 6.31) compared to the COX-2 plus PPI group. CONCLUSION: PPI use, but not other GI drug use, is a marker for increased susceptibility to ulcers among NSAID users. This risk of upper GI ulcers is increased in PPI users regardless of which NSAID is used (nonselective or COX-2-specific inhibitor). Although COX-2 use is associated with greater risk factors for upper GI ulcers due to channeling bias, COX-2 users have significantly fewer ulcers than equivalent nonselective NSAID users regardless of concomitant PPI utilization. PMID- 11908559 TI - Aerobic fitness, fatigue, and physical disability in systemic lupus erythematosus. AB - OBJECTIVE: To measure aerobic fitness, muscle strength, fatigue, and physical disability in patients with systemic lupus erythematosus (SLE). METHODS: Ninety three patients with SLE and 41 sedentary controls were recruited into the study. Aerobic fitness was assessed by monitoring peak and submaximal oxygen uptake, heart rate, duration of exercise, and perceived exertion during a treadmill walking test. Strength was measured using voluntary isometric quadriceps contraction. Symptomatic measures included physical and mental fatigue, mood, sleep, and functional incapacity. RESULTS: Compared to sedentary controls patients with SLE had significantly reduced levels of aerobic fitness (mean VO2peak SLE patients, 23.2 ml/kg/min vs controls, 29.6 ml/kg/min; p < 0.001) and reduced exercise capacity (mean exercise duration SLE patients, 10.4 min vs controls, 13.1 min; p < 0.001). The SLE patients also had reduced muscle strength (mean maximum voluntary quadriceps contraction SLE patients, 298 N vs controls, 376 N; p = 0.003). Resting lung function was also significantly worse in the SLE patients (mean FEV, SLE patients, 2.6 l vs controls, 2.9 l; p = 0.002). Fatigue (p < 0.001), depressed mood (p < 0.001), poor sleep quality (p < 0.001), and functional incapacity (p < 0.001) were all significantly greater in the SLE patients. Linear regression models suggested that physical disability correlated with aerobic fitness (p < 0.001), fatigue (p = 0.005), body mass index (p = 0.01), and depression (p = 0.05) and that fatigue correlated with depression (p < 0.001). CONCLUSION: Patients with SLE were less fit with reduced exercise capacity, reduced muscle strength, more fatigue, and greater disability compared to sedentary controls. Treatments developed to manage depression and improve aerobic fitness should be considered in the overall treatment of fatigue and disability in SLE. PMID- 11908560 TI - Fatigue in patients with systemic lupus erythematosus: lack of associations to serum cytokines, antiphospholipid antibodies, or other disease characteristics. AB - OBJECTIVE: To determine if fatigue in patients with systemic lupus erythematosus (SLE) is associated with levels of serum cytokines, antiphospholipid antibodies (aPL), or other disease features. METHODS: In a cross sectional study 57 Caucasian patients with SLE were subjected to clinical neurological examination and cerebral magnetic resonance imaging (MRI). Fatigue was evaluated by Fatigue Severity Scale (FSS) and disease activity by SLE Disease Activity Index (SLEDAI). Serum levels of tumor necrosis factor-alpha (TNF-alpha), interleukin 2 (IL-2), IL 6, IL-10, transforming growth factor-beta (TGF-beta), interferon-alpha (IFN alpha), anticardiolipin antibody (aCL) IgG and IgM, as well as anti-beta2 glycoprotein I antibody (anti-beta2-GPI) IgG and IgM were analyzed by ELISA. RESULTS: Four of 5 patients with SLE had fatigue (FSS score > or = 3). There were no associations between fatigue and any sociodemographic variables, medication for SLE, disease activity, cerebral infarcts, serum cytokines, aCL or beta2-GPI antibodies, or any routine hematological, biochemical, or immunological tests. CONCLUSION: Fatigue is a common phenomenon in patients with SLE. There is no association to disease activity or other markers of disease or inflammation. Fatigue is a complex phenomenon, and cytokine involvement in brain tissue not reflected by cytokine serum concentrations in this study cannot be excluded. Alternatively, psychosocial factors may well be the dominant predictor of fatigue in patients with SLE. PMID- 11908561 TI - Sicca symptoms and anti-SSA/Ro antibodies are common in mixed connective tissue disease. AB - OBJECTIVE: To examine a large well characterized cohort of patients with mixed connective tissue disease (MCTD) and determine longitudinally the prevalence of clinical and serologic features of Sjogren's syndrome in these patients. METHODS: Patients were followed longitudinally up to 30 years with systematic clinical and serologic analysis. Sera were analyzed for reactivity with SSA/Ro, SSB/La, and snRNP polypeptide U1-70kD and for anticardiolipin antibodies. RESULTS: Among a well characterized patient population with MCTD, 18/55 (32.7%) had antibodies to SSA/Ro, while 2/55 (3.6%) had antibodies to SSB/La, either initially or during the course of followup. All patients had antibodies to U1-70kD small nuclear ribonucleoprotein antigen. Sicca symptoms were common, occurring in 23/55 (41.8%) patients. Patients with MCTD who were anti-SSA/Ro positive had increased incidence of malar rash (p < 0.03) and photosensitivity (p < 0.001) compared to anti-SSA/Ro negative patients. CONCLUSION: Sicca symptoms are frequent in patients with MCTD, occurring in up to one-third of the patients studied. The presence of anti-SSA/Ro antibodies identifies a group of MCTD patients with a very high incidence of malar rash and photosensitivity. PMID- 11908562 TI - Longterm anticoagulation is preferable for patients with antiphospholipid antibody syndrome. result of a decision analysis. AB - OBJECTIVE: Patients with antiphospholipid antibody syndrome (APS) have a high risk for rethrombosis. Anticoagulation with warfarin and aspirin reduces the frequency of recurrences. No universally accepted approach regarding the duration and intensity of antithrombotic therapy exists. We investigated the best antithrombotic regimen for patients with APS after the first deep venous thrombosis (DVT). METHODS: We identified 6 anticoagulation regimens used in such patients, the rates of morbidity and mortality associated with bleeding, and the rates of recurrent thrombosis associated with APS by literature search. A decision tree was developed and the expected risks and benefits of each anticoagulation regimen were assessed at 2 different time points: at one year and again 4 years after the initial thrombosis. RESULTS: Based on the decision analysis, longterm warfarin alone at an international normalization ratio (INR) between 3.0 and 4.0 had the highest expected utility of the 6 antithrombotic regimens, both one year and 4 years after the initial venous thrombotic event. Short term anticoagulation for only 6 months is less beneficial. Combination therapy of warfarin and aspirin (ASA) does not offer an improvement in the expected utility over warfarin alone. CONCLUSION: Although the applicability of this analysis to clinical decision-making is not entirely clear, patients with APS presenting with DVT appear to benefit from longterm warfarin (INR 3.0-4.0) that may be superior to warfarin (INR 2.0-3.0). Short term warfarin therapy seems to be less beneficial and the use of ASA does not offer a clear additional benefit. Randomized controlled trials are needed to provide a better basis for recommendations for the treatment APS. PMID- 11908563 TI - Opportunistic infections are preceded by a rapid fall in antineutrophil cytoplasmic antibody (ANCA) titer in patients with ANCA associated vasculitis. AB - OBJECTIVE: To study the clinical course and changes in antineutrophil cytoplasmic antibody (ANCA) titers in patients with ANCA associated vasculitis (AAV) who developed opportunistic infections. METHODS: Among the patients with AAV tested in the Immunopathology Laboratory at the Massachusetts General Hospital between 1989 and 1998, all patients who experienced opportunistic infections (n = 16) were included. We retrospectively studied their clinical features and examined the relationship between changes in ANCA titer and the onset of the opportunistic infections. ANCA titers were measured by antigen-specific ELISA. RESULTS: Of the 16 AAV patients with opportunistic infection, 15 had no evidence for active vasculitis at the time of the infection. Among these 15 patients, opportunistic infections were associated with a steep fall in ANCA titers. There was no consistent pattern of change in C-reactive protein levels. In 7 patients, the immunosuppressive regimen was increased for new clinical findings shortly before the diagnosis of an opportunistic infection, despite the absence of histologic documentation of active vasculitis. Three of these 7 patients died. One patient, who did not experience a significant fall in ANCA titer. i.e., less than 4-fold from his prior peak, was simultaneously found to have Pneumocystis carinii pneumonia and biopsy proven active vasculitis. CONCLUSION: Our data strongly suggest that opportunistic infections in patients with AAV are associated with negative or rapidly falling ANCA titers. Therefore, changes in ANCA titer can help distinguish opportunistic infections from vasculitis flares when patients with AAV present with indeterminate clinical findings. PMID- 11908564 TI - Prevalence of ankylosing spondylitis and other spondyloarthropathies among patients with inflammatory bowel disease: a population study (the IBSEN study). AB - OBJECTIVE: To study the occurrence of spondyloarthropathies (SpA) in patients with inflammatory bowel disease (IBD) seen 6 years after IBD diagnosis. METHODS: In a population based cohort of 654 patients with IBD, 521 patients (80%) were investigated, which included a complete rheumatological examination. Radiographs of the sacroiliac joints and lumbar spine were performed in 406 of these patients (78%). The development of SpA was analyzed with regard to the presence of HLA B27, duration of IBD symptoms, and the extent of intestinal inflammation. RESULTS: The occurrence of ankylosing spondylitis (AS) was 2.6% in ulcerative colitis and 6% in Crohn's disease (p = 0.08), yielding an overall prevalence of 3.7% in IBD. No correlation between localization or extent of the intestinal inflammation and presence of AS was found. HLA-B27 was present in 73% of cases with AS. The overall prevalence of SpA was 22%. Inflammatory back pain without AS (IBP) was found in 18% of the patients. Typical features of SpA were rare, while fibromyalgia was common in IBP, indicating that IBP is not a precursor or manifestation of SpA in patients with IBD. The prevalence of radiological sacroiliitis without clinical features of SpA was 2.0%. CONCLUSION: AS occurred frequently in patients with newly diagnosed IBD. IBP did not seem to predispose to AS or other forms of SpA. The overall prevalence of SpA was 22%, whereas the prevalence of asymptomatic radiological sacroiliitis was low. PMID- 11908565 TI - Bone mineral density, calcaneal ultrasound, and bone turnover markers in women with ankylosing spondylitis. AB - OBJECTIVE: To assess bone mineral density (BMD) by dual energy x-ray absorptiometry (DEXA) and calcaneal quantitative ultrasound (QUS) in a cohort of pre- and postmenopausal women with ankylosing spondylitis (AS), and to determine any relationships with markers of bone turnover and disease activity or severity. METHODS: Fifty premenopausal and 16 postmenopausal women with AS were studied. Clinical and radiological status was assessed by the Bath AS Disease Activity Index (BASDAI), Bath AS Functional Index (BASFI), Bath AS Metrology Index (BASMI), and Bath AS Radiology Index (BASRI). BMD of the hip and spine was measured by DEXA, and QUS measured at the heel. Serum osteocalcin (OC), bone specific alkaline phosphatase (BALP), urinary D-pyridinoline crosslinks (D-PYR), and C-reactive protein (CRP) were assayed. RESULTS: Women with AS (n = 66) had reduced BMD at the hip compared to age and sex matched controls (n = 132). The mean t scores were -1.1 and -2.0, and z scores -0.4 and -0.37, for pre- and postmenopausal women, respectively. Four (6%) had osteoporosis and 34 (52%) had osteopenia according to the WHO definitions. Using a multiple regression model, femoral neck BMD was found to be significantly affected by age, body mass index, and the sacroiliac radiographic score. There were no significant correlations of BMD with disease duration or disease activity. QUS measures did not correlate with DEXA measures of BMD. Women with AS had significantly lower markers of bone formation, OC and BALP, and a trend to higher D-PYR than controls. Serum OC levels correlated negatively with femoral neck BMD, whereas D-PYR correlated with CRP levels. CONCLUSION: Women with AS have reduced hip BMD, 0.39 SD below age and sex matched controls. Bone turnover in women with AS is characterized by low OC and BALP. PMID- 11908566 TI - Interleukin 1beta (IL-1beta), IL-10, tumor necrosis factor-alpha, and cellular proliferation index in peripheral blood mononuclear cells in patients with ankylosing spondylitis. AB - OBJECTIVE: To evaluate cytokine production and cellular proliferation index (CPI) in peripheral blood mononuclear cells (PBMC) of patients with ankylosing spondylitis (AS), and their association with clinical variables. METHODS: In a cross sectional study we compared the production of tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), and IL-10 and CPI in response to phytohemagglutinin (PHA) in PBMC of 27 patients with AS and 24 healthy controls. We also assessed clinical characteristics including disease activity index (BASDAI) and functional index (BASFI). RESULTS: Levels of IL-1beta were higher in patients with AS (median 242 pg/ml) than in controls (median 65 pg/ml); p = 0.002. No differences were observed in median levels of TNF-alpha or IL-10 between AS and controls. Patients had a reduction in CPI (1.2 in AS vs 1.8 in controls; p < 0.001). A positive correlation was observed between IL-10 production and age (rho = 0.34, p = 0.01). A borderline negative correlation was observed between CPI and age (rho = -0.26, p = 0.07). CONCLUSION: Patients with AS had high production of IL-1beta compared with controls and a poor response in CPI. These findings may explain the lack of response for microbial antigens mediated by the innate immune response. PMID- 11908567 TI - Birth order and ankylosing spondylitis: no increased risk of developing ankylosing spondylitis among first-born children. AB - OBJECTIVE: In the HLA-B27 transgenic mouse model the first litters have been shown to have a higher percentage of diseased offspring than later litters. First born children (n = 162) have also been shown to have a higher risk of ankylosing spondylitis (AS) than later-born children. We examined this effect of birth order using similar methods but larger numbers. METHODS: Patients from the Bath AS database (n = 4517; M:F = 2.5:1) were examined according to position of birth within the family. Chi-squared analysis was used to examine if AS was more prevalent among first-born than later-born children. RESULTS: The first-born child was not significantly more likely to have AS than later-born children (p = 0.295). [Observed compared to expected: 1607 (36%) compared to 1641.13 (36%) for first-born children and 2910 (64%) compared to 2876.3 (64%) for later-born children, respectively.] There was no biological gradient (i.e., inverse correlation between birth order and disease risk). CONCLUSION: There was no statistically significant effect of birth order based on our data. Findings suggesting a birth order effect may be skewed, as it is possible that those parents who do have AS will be less likely to have a large family and yet it is their offspring who will be at greatest risk of developing disease. This will affect the data, as those children born into a large family (i.e., high birth order children) will be at a lower risk of AS than any child born into a small but family-history-positive unit. PMID- 11908568 TI - Familial mediterranean fever and Behcet's disease--are they associated? AB - OBJECTIVE: To test whether the coexistence of familial Mediterranean fever (FMF) and Behcet's disease (BD) is more frequent than expected and whether each disease affects the severity of the other. METHODS: We screened 353 charts of patients with FMF to detect individuals with concomitant BD. Of these, 152 patients with FMF over the age of 18 years were also interviewed and examined specifically. We also studied 53 patients with BD, looking for FMF and for their MEFV mutations. We compared BD patients with MEFV mutations to those without them. RESULTS: None of 353 patients with FMF was found to have concomitant BD. Sixteen patients with BD bore MEFV mutations, 2 of whom were symptomatic homozygotes and had concomitant FMF. No patient with BD with a single MEFV mutation had FMF. Both BD groups (with or without MEFV mutations) were similar in their clinical manifestations and disease course. CONCLUSION: BD and FMF are 2 separate entities that have a mild trend toward a higher than expected association. However, there was no mutual effect of FMF on BD or vice versa. PMID- 11908570 TI - Correlation between body composition and efficacy of lateral wedged insoles for medial compartment osteoarthritis of the knee. AB - OBJECTIVE: To investigate anthropometric measures that closely correlate with symptomatic relief of osteoarthritis (OA) of the knee in response to lateral wedged insole use. METHODS: Seventy-one patients with medial compartment knee OA were treated with insoles with subtalar strapping or insoles with talonavicular strapping. Randomization was performed according to birth date. The following variables were evaluated: age, disease duration, Kellgren-Lawrence radiographic stage, body mass index, percent body fat, waist to hip ratio, lower extremity lean body mass (L-LBM) per body weight, and radiographic femorotibial angle at baseline. The trial lasted 8 weeks. The correlation between each variable and the remission score (delta score) using the Lequesne index of severity was analyzed. RESULTS: In the subtalar strapping group (n = 34), delta score of knee OA was more strongly associated with age (p = 0.004, r = 0.48) than other variables. A significant correlation was also observed between L-LBM per body weight and delta score (p = 0.041, r = -0.36) in the subtalar strapping group. No other variables significantly correlated with the delta score in the subtalar strapping groups. No variable significantly correlated with the ? score in the talonavicular strapping group (n = 37). CONCLUSION: We previously reported that use of insoles with subtalar strapping leads to valgus realignment of the femorotibial angle in patients with knee OA with varus deformity, and it may have a similar therapeutic effect to that of high tibial osteotomy. These data suggest that the insole with subtalar strapping is more efficacious for younger patients and those with a higher L-LBM per body weight, and less efficacious for older patients with sarcopenia. PMID- 11908569 TI - Endothelial nitric oxide synthase gene polymorphisms in Behcet's disease. AB - OBJECTIVE: To analyze potential associations of Glu-Asp298 polymorphism in exon 7 and 4 a/b polymorphism in intron 4 of the endothelial nitric oxide synthase (eNOS) gene with susceptibility for Behcet's disease (BD). METHODS: Seventy-three consecutive Italian patients who satisfied the International Study Group criteria for BD and 135 healthy blood donor controls from the same geographic area were genotyped by polymerase chain reaction and allele-specific oligonucleotide techniques for eNOS polymorphisms in exon 7 and in intron 4. RESULTS: The distribution of the Glu-Asp298 genotype differed significantly between BD patients and controls (p(corr) = 0.00009). Allele Asp298 was significantly more frequent in BD patients than in controls (p(corr) = 0.0006, OR 2.1, 95% CI 1.5 3.3). The distribution of 4 a/b genotype was similar in patients and controls. CONCLUSION: Our findings show that Glu-Asp298 polymorphism of eNOS gene is associated with BD susceptibility. PMID- 11908571 TI - Synthesis of interleukin 1beta, tumor necrosis factor-alpha, and interstitial collagenase (MMP-1) is eicosanoid dependent in human osteoarthritis synovial membrane explants: interactions with antiinflammatory cytokines. AB - OBJECTIVE: To determine the level of leukotriene B4 (LTB4) synthesized and released by synovium of patients with osteoarthritis (OA), and to study the role of lipoxygenase (LO)/cyclooxygenase (COX) products on proinflammatory cytokine and interstitial collagenase (MMP-1) synthesis. METHODS: Human OA synovial explants were cultured in the presence of lipopolysaccharide (L) and the ionophores ionomycin (I) and thapsigargin (T) (LIT) for 72 h at 37 degrees C, and LTB4 released into the culture medium was measured in the absence or presence of a COX-2-specific inhibitor, NS-398, or the 5-LO activating protein inhibitor Bay x-1005. Increasing concentrations of LTB4 (10(-9) to 10(-6) M) were incubated with explants for 24 h at 37 degrees C, and interleukin 1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in the conditioned medium were quantitated by ELISA. The effect of endogenous eicosanoids on basal and induced levels of IL-1beta, TNF-alpha, and MMP-1 synthesis was examined by incubating explants in the presence of NS-398 and Bay-x-1005. The effect of antiinflammatory cytokines rhIL-4, IL-10, and IL-13 on basal and LTB4 dependent stimulation of IL 1beta/TNF-alpha synthesis was studied under titration conditions. RESULTS: Physiologically relevant concentrations (10(-10) to 10(-9) mol/l) of LTB4 were produced in the presence of LIT. Bay-x-1005 abrogated LTB4 release, while NS-398 was without effect. LTB4 stimulated IL-1beta and TNF-alpha synthesis with an EC50 of 190 +/- 35 and 45 +/- 9 nmol/l, respectively. Significant concentrations of IL 1beta and TNF-alpha were released (100-200 and 500-600 pg/ml, respectively). Basal and LIT induced IL-1beta and TNF-alpha production were inhibited by Bay-x 1005 in a dose dependent manner, while the addition of NS-398 caused a potent stimulatory effect. The preferential COX-2 inhibitor also induced MMP-1 synthesis in a manner essentially identical to the proinflammatory cytokines. The antiinflammatory cytokine IL-4 blocked LTB4 dependent stimulation of IL-1beta and TNF-alpha synthesis. In contrast, IL-10 markedly stimulated both cytokines when incubated alone or in the presence of LTB4 where the effect was additive. CONCLUSION: Endogenous and locally produced eicosanoids regulate proinflammatory cytokine and MMP-1 synthesis under basal and stimulated conditions in vitro, with leukotrienes and prostaglandins having opposite effects in general. The clinical use of antiinflammatory drugs that inhibit eicosanoid synthesis requires an appreciation of their relative capacity to inhibit LO/COX in order to predict their effect on the synthesis of proinflammatory cytokines and matrix metalloproteases. IL-10 stimulated proinflammatory cytokine synthesis in our ex vivo culture system. PMID- 11908572 TI - Compartment differences in knee cartilage volume in healthy adults. AB - OBJECTIVE: It is unclear why there are compartmental differences in the risk of knee osteoarthritis (OA). We investigated whether there are compartment differences in the volume of knee cartilage in healthy persons and identified determinants of medial and lateral tibial cartilage volume. METHODS: A total of 166 healthy persons (age range 21-79 yrs, 58% female) with clinically and structurally normal knees were examined. Thickness and volume were determined for the medial and lateral tibial articular cartilages by processing images acquired in the sagittal plane using T1 weighted fat saturated magnetic resonance imaging on an independent work station. RESULTS: In every subject, the lateral tibial cartilage was thicker than medial cartilage (mean 6.43 +/- SD 1.25 mm vs 4.49 +/- 0.81 mm; p < 0.001), and the volume of cartilage was greater (2.34 +/- 0.70 ml vs 1.82 +/- 0.56 ml; p < 0.001). This effect persisted when values for men and women were analyzed separately. Both medial and lateral tibial cartilage volume and thickness were greater in men compared to women, independent of body mass index and bone size. The reduction in medial and lateral tibial cartilage volume and thickness was inversely related to the current level of physical activity. CONCLUSION: This study supports the knee compartment differences in cartilage volume recently reported in children. It is likely these differences are maintained throughout life. The possibility that the amount of knee cartilage in an individual is a risk factor for OA now needs to be tested in longitudinal studies. PMID- 11908573 TI - Sports injury, occupational physical activity, joint laxity, and meniscal damage. AB - OBJECTIVE: To investigate the risk factors for meniscal damage, an important determinant of knee osteoarthritis. METHODS: We studied 243 men and women aged 20 59 years in whom the diagnosis of a meniscal tear was confirmed for the first time at arthroscopy, over a 25 month period, in 2 British hospitals. Each case was compared with one or 2 community controls, matched by age and sex, who were registered with the same general practitioner. Information on exposure to risk factors was obtained by a structured questionnaire and physical examination. RESULTS: Meniscal tear was strongly associated with participation in sports during the 12 months preceding the onset of symptoms; the risk was particularly high for soccer (OR 3.7; 95% CI 2.1-6.6). Higher body mass index and occupational kneeling (OR 3.8; 95% CI 1.3-11.0) and squatting (OR 2.9; 95% CI 1.0-8.0) were associated with an increased risk of degenerative meniscal lesions, after adjustment for social class, joint laxity, and sports participation. Joint laxity was associated with degenerative meniscal lesions independently of occupational physical activity, sports, and obesity. CONCLUSION: Our results confirm the importance of sporting activities entailing knee torsion in acute meniscal tear. They also point to a role for occupational activity, adiposity, and joint laxity in the pathogenesis of degenerative meniscal lesions. Modifying these mechanical risk factors may serve to reduce the risk of meniscal injury and may also help to prevent later knee osteoarthritis. PMID- 11908574 TI - Neck and upper limb pain: more pain is associated with psychological distress and consultation rate in primary care. AB - OBJECTIVE: To investigate the association between the extent of pain and the severity of psychological distress in neck and upper limb pain, and to establish whether extent of pain is associated with consultation frequency in primary care. METHODS: The study population was selected from responders to a general health survey conducted in a general practice in North Staffordshire, UK. Responders indicating pain in the neck or upper limb area were included. The survey included the Hospital Anxiety and Depression Scale (HADS). Consultation data were retrieved for a period of 12 months following the survey. RESULTS: A total of 867 responders had experienced neck-upper limb pain in the month preceding the survey (33% of all responders). Responders with more generalized pain within the neck upper limb area had significantly higher HADS scores compared to responders with pain in one area only, particularly for depression (median scores 5 vs 3 points). Annual consultation frequency was also higher among responders with generalized pain [adjusted OR for high consultation frequency (> or = 7 visits vs 0-2 visits) 1.6, 95% CI 1.1 to 2.4]. When the analysis was restricted to consultations specifically related to neck-upper limb pain, the association between extent of pain and consultation frequency was weak and not statistically significant. CONCLUSION: Our survey revealed a significant association between extent of pain in the neck-upper limb area and psychological distress, although scores for anxiety and depression were generally low, with only a small proportion of responders reporting moderate or severe symptoms. Responders with both generalized pain and depressive symptoms were more likely to consult their family doctor, but not specifically for musculoskeletal pain. These results confirm the hypothesis that general psychological well being rather than specific somatic symptoms predict consultation frequency. PMID- 11908575 TI - The high prevalence of pathologic calcium crystals in pre-operative knees. AB - OBJECTIVE: Calcium pyrophosphate dihydrate (CPPD) and basic calcium phosphate (BCP) crystals are important in the pathogenesis of osteoarthritis (OA) but are under recognized even in end stage disease. We determined the prevalence of these calcium crystals in synovial fluid (SF) of persons undergoing total knee arthroplasty for degenerative arthritis. METHODS: SF samples were obtained from 53 knee joints undergoing total arthroplasty for a pre-operative diagnosis of OA. SF were analyzed via compensated light microscopy for CPPD crystals and a semiquantitative radiometric assay for BCP crystals. Fifty pre-operative radiographs were analyzed and graded according to the scale of Kellgren and Lawrence. RESULTS: Patients had an average age of 70 years at the time of surgery. CPPD and/or BCP crystals were identified in 60% of SF. Overall radiographic scores correlated with mean concentrations of BCP crystals. Higher mean radiographic scores correlated with the presence of calcium-containing crystals of either type in SF Radiographic chondrocalcinosis was identified in only 31% of those with SF CPPD. CONCLUSIONS: Pathologic calcium crystals were present in a majority of SF at the time of total knee arthroplasty. Intraoperative SF analysis could conveniently identify pathologic calcium crystals providing information that may be relevant to the future care of the patient's replaced joint and that of other joints. This information could also potentially aid in predicting the likelihood of the need for contralateral total knee arthroplasty. PMID- 11908576 TI - A randomized clinical trial comparing fitness and biofeedback training versus basic treatment in patients with fibromyalgia. AB - OBJECTIVE: To compare the therapeutic effects of physical fitness training or biofeedback training with the results of usual care in patients with fibromyalgia (FM). METHODS: One hundred forty-three female patients with FM (American College of Rheumatology criteria) were randomized into 3 groups: a fitness program (n = 58), biofeedback training (n = 56), or controls (n = 29). Half the patients in the active treatment groups also received an educational program aimed at improving compliance. Assessments were done at baseline and after 24 weeks. The primary outcome was pain [visual analog scale (VAS)]. Other endpoints were the number of tender points, total myalgic score (dolorimetry), physical fitness, functional ability (Arthritis Impact Measurement Scale and Sickness Impact Profile), psychological distress (Symptom Checklist-90-Revised), patient global assessment (5 point scale), and general fatigue (VAS). RESULTS: Baseline scores were similar in all 3 groups. Altogether 25 (17.5%) patients dropped out; they were similarly distributed over all groups: 14 patients after randomization and 11 (8%) during the study. A true high impact level for fitness training was not attained by any patient. After treatment, no significant differences in change scores of any outcome were found between the groups (ANOVA, p > 0.05). All outcome measures showed large variations intra- and interindividually. The educational program did not result in higher compliance with training sessions (62% vs 71%). Analysis of the subgroup of subjects with a high attendance rate (> 67%) also showed no improvement. CONCLUSION: In terms of training intensity and maximal heart rates, the high impact fitness intervention had a low impact benefit. Therefore effectiveness of high impact physical fitness training cannot be demonstrated. Thus compared to usual care, the fitness training (i.e., low impact) and biofeedback training had no clear beneficial effects on objective or subjective patient outcomes in patients with FM. PMID- 11908577 TI - High or low intensity aerobic fitness training in fibromyalgia: does it matter? AB - OBJECTIVE: To determine the efficacy of training in fibromyalgia (FM), we compared the effects of high intensity fitness training (HIF) and low intensity fitness training (LIF). METHODS: Thirty-seven female patients with FM were randomly allocated to either a HIF group (n = 19) or a LIF group (n = 18). Four patients (1 HIF group, 3 LIF group) refused to participate after randomization but before the start of the intervention. They were excluded from the analysis. Assessments were performed at baseline and after 20 weeks of HIF or LIF. The primary outcome was patient's global assessment [on 100 mm visual analog scale (VAS)]. Secondary endpoints were pain, number of tender points, total myalgic score, physical fitness, health status, and psychological distress. RESULTS: One patient in the HIF group (n = 18) and 2 in the LIF group (n = 15) stopped training sessions during the course of the study. Nine of 18 patients in the HIF group compared to 8 of 15 patients in the LIF group achieved a participation rate of 67% or more. Most important reasons for nonadherence were postexercise pain and fatigue, time consumption, and stress. The VAS for global well being improved slightly from 64 to 56 mm in the HIF group, and did not change in the LIF group (58 to 61 mm) (p = 0.07). The Wmax (physical fitness) changed modestly from 110 to 123 watt in the HIF group, and from 97 to 103 watt in the LIF group (p = 0.3). VAS for pain increased from 53 to 64 mm in the HIF group and from 52 to 54 mm in the LIF group. The large standard deviations around mean change in global assessments, number of tender points, total myalgic score, and psychological distress (by SCL-90) severely influenced the power to detect within- and between group differences. Analysis limited to those patients who accomplished a high attendance rate (> 67%) showed similar results. CONCLUSION: High intensity physical fitness training compared to low intensity physical fitness training leads to only modest improvements in physical fitness and general well being in patients with FM, and does not positively affect psychological status and general health. PMID- 11908578 TI - Chronic widespread musculoskeletal pain with or without fibromyalgia: psychological distress in a representative community adult sample. AB - OBJECTIVE: To estimate the severity of depression, anxiety, and other symptoms of psychological distress in a representative general population sample of fibromyalgia (FM) cases (FC) compared to pain controls (PC), and to identify strong correlates of depression and anxiety. METHODS: We compared the severity of depression, anxiety, and other symptoms of psychological distress between 2 representative community samples: (1) 74 confirmed FC, and (2) 48 adults with chronic widespread pain (PC) who did not meet the 1990 ACR criteria for FM. Psychological distress was measured using the Centre for Epidemiological Studies Depression (CES-D) Scale, the State-Trait Anxiety Inventory (STAI), and other measures of psychological distress from the literature. Using cutoff scores for CES-D and trait anxiety, we compared demographic and clinical characteristics in those above and below each cutoff score. Simple linear regression was performed to identify factors strongly and independently correlated with depression and trait anxiety. RESULTS: Compared to PC, FC were more symptomatic on virtually all measures of psychological distress. Similarly, individuals who scored above cutoff scores for depression and anxiety had more physical symptoms and had poorer function than those below. Depression and trait anxiety were highly correlated (r = 0.86). In a simple regression model, the best predictors for both depression and trait anxiety were the total number of symptoms and a physical disability score. CONCLUSION: Depression and anxiety are common and frequently severe even among community cases of FM. PMID- 11908579 TI - Tender point scores and their relations to signs of mobility, symptoms, and disability in female home care personnel and the prevalence of fibromyalgia syndrome. AB - OBJECTIVE: In this study of female home care personnel employed in a municipality (n = 643; participation rate 94%) we investigated (1) the prevalence of tender points and fibromyalgia (FM); (2) the relationships between tender point score and other signs and symptoms; (3) if subgroups based on the tender point score differed with respect to signs, symptoms, disability, and health related quality of life; and (4) signs that showed the strongest intercorrelations with disability and health. METHODS: The following variables were registered: (1) Signs: joint mobility, spinal posture and mobility, tender points, and segmental mobility and pain provocation at L4-S1 levels of the low back. (2) SYMPTOMS: pain and pain intensity and other symptoms. (3) Disability (i.e., self-rated reduced capacity for everyday activities and employment) and health: 3 indices and sick leave. RESULTS: The tender point score correlated with the number of pain regions and the pain intensities, and the amount of other symptoms, sick leave, and disability. Tender point score was the strongest regressor of the investigated signs in regression of the 2 disability indices. Segmental pain showed the strongest correlation with tender point score. Three subgroups identified by tender point score showed significant differences in segmental pain, prevalence and intensity of different symptoms, disability, and health related quality of life. The prevalence of FM was 2.0%. CONCLUSION: Tender point score together with different symptoms showed relatively strong correlations with disability. A relatively high prevalence of FM was found in occupationally active female home care personnel. PMID- 11908580 TI - Pain and disability, perceptions and beliefs of a rural Indian population: A WHO ILAR COPCORD study. WHO-International League of Associations for Rheumatology. Community Oriented Program for Control of Rheumatic Diseases. AB - OBJECTIVE: The WHO-ILAR Community Oriented Program for Control of Rheumatic Diseases (COPCORD) primarily aims to estimate the burden of rheumatic musculoskeletal symptoms/disorders (RMS). We investigated data on pain and disability, perceptions and beliefs in the first rural community based COPCORD study in India. METHODS: A total of 4092 adults were interviewed (response rate 89%) in a population survey (Stage 1) in Bhigwan village in 1996 using modified COPCORD core questionnaires. Twenty-one trained volunteers completed the survey in 5 weeks. Those reporting RMS were identified (Phase 1) to complete a self evaluation questionnaire (Phase 2) prior to rheumatological evaluation (Phase 3). Phase 2 included questions on perceptions and beliefs regarding pain, effect on life, work and socioeconomic factors, disability, and therapy; only the moderate and severe grades were considered significant. Patients marked their pain sites on a manikin during the presurvey week. A validated modified Health Assessment Questionnaire disability index (HAQDI) in the local language evaluated functional disability. RESULTS: RMS were the predominant ailments reported by 746 adult villagers (18.2%; 95% CI 17.1, 19.2). Moderate pain of > 2 years' duration was reported by almost 60% of RMS patients. Neck (6%), lumbar (11.4%), shoulder (7.4%), elbow (6.5%), wrist (6.4%), hand (6.1%), knee (13.2%), calf (6.6%), and ankle (6.5%) were the common painful sites, predominantly in women; 91%, 89%, and 31% with RMS reported a significant grade of pain, RMS illness, and disturbed sleep, respectively. In the age group 25-54 years, 21% of those with RMS perceived a significant effect on work ability, while less than 20% of those with RMS admitted a similar effect on their personal life (including finances). About 10% with RMS had ceased to work because of RMS. Among RMS subjects 21% scored a significant HAQDI, but many more reported significant difficulty (HAQ) in the individual items of walking, hygiene (squatting), arising (from sitting cross legged), reaching, and occupational/household chores; this corresponded to the dominant pain sites in low back and lower limbs. Oral tobacco use was reported to be significantly greater (p < 0.001) in the RMS patients. Past trauma was recalled by 23% of patients, and many connected this to their RMS. Modern medicines were consumed by 55% of patients with RMS. Among patients, 86% and 65% expected "pain relief" and "cure," respectively, from their doctor; 23% of patients wanted greater sympathy and attention. However, 21% of patients had never visited a doctor and were only identified by the COPCORD study. CONCLUSION: The findings of this study (1) demonstrate that RMS, although a predominant ailment, has a modest effect on daily living in most subjects with RMS; (2) indicate there is inconsistency between the measures of pain and disability (using HAQ) and their effects; (3) describe the beliefs and expectations of the community. Based on the data and community support, the COPCORD has been continued for Stages II and III, especially with a view to health education. PMID- 11908581 TI - Health care provision in pediatric rheumatology in Germany--national rheumatologic database. AB - OBJECTIVE: To describe the health care provision for children and adolescents with chronic arthritides in Germany in 1998. METHODS: Data were analyzed from the German pediatric rheumatologic database of the year 1998. It contains clinical and patient questionnaire data for 2488 patients with rheumatic diseases seen at 18 pediatric rheumatology units. RESULTS: A total of 1811 of all patients recorded in the database had chronic arthritides--931 with juvenile chronic arthritis, 86 with juvenile spondyloarthropathy, and 65 with juvenile psoriatic arthritis were considered in the analysis. These patients seen by pediatric rheumatologists had a median age of 10 years and a median disease duration of 4 years. The majority were being treated at pediatric rheumatology disease centers and at universities. Nonsteroidal antiinflammatory drugs were the most commonly used drugs for all forms of chronic arthritides. Almost half the patients with chronic arthritides received disease modifying antirheumatic drugs, with methotrexate the most frequently prescribed agent. While the majority of patients reported having physiotherapy, low prescription rates were noted for comprehensive measures such as occupational therapy and patient education. Only a few patients showed severe functional limitation, 2% of them being rated in Steinbrocker class III or higher. While the patients' functional limitation correlated with disease activity, neither disease duration nor sex, arthritis subgroup nor time span to the first visit at the rheumatology unit had any relevant influence on functional status. CONCLUSION: The data reveal the spectrum of patients with chronic arthritides seen by German pediatric rheumatologists, as well as the treatment patterns of their physicians. PMID- 11908582 TI - Ultrasonography of the hip in the evaluation of children with seronegative juvenile rheumatoid arthritis. AB - OBJECTIVE: To find an objective measure of hip joint effusion with ultrasound (US) in patients with juvenile rheumatoid arthritis (JRA). METHODS: The hip joints of 24 children with JRA were evaluated with US. All patients were negative for rheumatoid factor and antinuclear antibodies. Patients with unilateral or bilateral hip pain, swelling, or limitation of range of motion were included. In each hip, the distance from the femoral neck to joint capsule was measured. Values were compared to measurements in a control group of 24 children with no history of hip joint or rheumatic disease. Statistically significant differences between the 2 groups were analyzed by t test. Two standard deviations above the control group mean was used as the standard for an effusion. RESULTS: There was a statistically significant difference in US joint space between the children with JRA and the control subjects (p < 0.001). The mean in the control group was 0.43 cm and the mean in the JRA group was 0.60 cm. A distance of 0.59 cm from femoral neck to joint capsule was determined to be consistent with an effusion. Using this standard, 71% of the children with JRA had effusion in at least one hip, and 25% had effusion bilaterally. No control subjects had measurements above this level. CONCLUSION: Ultrasonography is effective in the evaluation of hip joint involvement in patients with JRA, and may be useful in facilitating the diagnosis, classification, and followup of this illness. PMID- 11908583 TI - Neuropsychiatric systemic lupus erythematosus presenting as amyotrophic lateral sclerosis. AB - A 61-year-old woman with a history of photosensitive dermatitis and recurrent mouth ulcers presented with progressive weakness typical of amyotrophic lateral sclerosis (ALS), and subsequently underwent extensive neurologic and rheumatologic testing. We investigated whether ALS-like motor neuron disease associated with a positive antinuclear antibody (ANA) is really ALS or rather neuropsychiatric systemic lupus erythematosus (NPSLE). On neurologic evaluation, she had prominent bulbar involvement with dysarthria and dysphagia associated with profound lingual fasciculations and a denervating pattern on electromyogram. MRI showed no evidence of cerebral ischemia. Laboratory studies revealed a positive ANA (1:2560 titer), positive antiphospholipid antibodies (GPL and MPL), circulating lupus anticoagulant, and depressed C3 and C4. Repeat MRI studies at 4 and 11 mo revealed an evolving infarct in the paramedian pons consistent with the presence of NPSLE. Therapy was initiated with corticosteroids and intravenous cyclophosphamide, and the neurologic condition did not improve, but also did not progress inexorably as would be expected with ALS. NPSLE, presumably through the mechanism of ischemic vasculopathy, may present as motor neuron disease clinically indistinguishable from ALS. PMID- 11908584 TI - Infliximab treatment in a patient with rheumatoid arthritis on hemodialysis. AB - We describe a 60-year-old woman with active rheumatoid arthritis (RA) and endstage renal disease secondary to hypertensive nephrosclerosis undergoing hemodialysis. She had tried multiple antirheumatic medications; however, their usefulness was limited due to toxic side effects or lack of efficacy. She was then treated with chimeric antitumor necrosis factor monoclonal antibody (infliximab), which resulted in immediate improvement in clinical and laboratory measures. After about 2 years of therapy, no side effects have been observed. This report expands the spectrum of infliximab to include RA patients with renal insufficiency. PMID- 11908585 TI - Neuropathic shoulder arthropathy associated with syringomyelia and Arnold-Chiari malformation (type I). PMID- 11908586 TI - Simple analgesics versus NSAID. PMID- 11908587 TI - Autoinimune disease among teachers. PMID- 11908588 TI - Breast implants and fibromyalgia. PMID- 11908589 TI - The death of Mozart. PMID- 11908590 TI - Do resemblance ratings measure the accuracy of facial approximations? AB - Since forensic facial approximations are used to promote recognition of a deceased person, an accurate forensic facial approximation (FFA) should be easily recognized as the person to whom the skull belonged (target individual). However, the accuracy of FFAs has been previously assessed by the direct comparison of an FFA to the corresponding target individual for similarity (i.e., a resemblance rating). Resemblance ratings may not indicate a facial approximation's accuracy since the resemblance of non-target individuals is not accounted for. This experiment tests the validity of using resemblance ratings to assess the accuracy of FFAs. The study indicates that there is no statistically significant difference between: (a) resemblance ratings of FFAs to target individuals and (b) resemblance ratings of FFAs to individuals incorrectly identified as the target individual. It is concluded that it is not possible from resemblance ratings to determine the accuracy and/or quality of a facial approximation since a non target individual may receive a resemblance rating equal to, or higher than, the target individual. PMID- 11908591 TI - Development of fluorescent markers using polycyclic aromatic hydrocarbons with vaseline. AB - Identifiable fluorescent markers were developed as tracers to tail suspects using phenanthrene, anthracene, fluoranthene, pyrene, perylene, and coronene in vaseline. Vaseline was used as a carrier of the marker. Of the six compounds in the vaseline, perylene and fluoranthene were readily observed under ultraviolet (UV) light at a wavelength of 365 nm. All six compounds were identified selectively and sensitively without interference of vaseline using a high performance liquid chromatograph (HPLC) with a fluorescence detector. The detection limit was much less than 1 ng, corresponding to that of the observation behavior under UV light. The results showed that each component with vaseline was more effective than the individual component for the delay in degradation. The case examples of the fluorescent markers are shown. PMID- 11908592 TI - Evaluation of some oxygen, sulfur, and selenium substituted ninhydrin analogues, nitrophenylninhydrin and benzo[f]furoninhydrin. AB - Six ninhydrin analogues containing oxygen, sulfur, and selenium substituents at the C-5 position, 5-(4-nitrophenyl)ninhydrin, and benzo[f]furoninhydrin were evaluated as fingerprint development reagents. The analogues all showed good fingerprint color development but were not superior to ninhydrin in this respect. The benzo[f]furoninhydrin complex was strongly luminescent at room temperature following zinc complexation, while the remaining analogues required cooling to 196 degrees C to produce optimum luminescence. The benzo[f]furo, nitrophenyl, and methyl selenide analogues showed the best potential as fingerprint reagents with the benzo[f]furo analogue comparing favorably with DFO. PMID- 11908593 TI - Associating gunpowder and residues from commercial ammunition using compositional analysis. AB - Qualitatively identifying and quantitatively determining the additives in smokeless gunpowder to calculate a numerical propellant to stabilizer (P/S) ratio is a new approach to associate handgun-fired organic gunshot residues (OGSR) with unfired powder. In past work, the P/S values of handgun OGSR and cartridges loaded with known gunpowders were evaluated. In this study, gunpowder and residue samples were obtained from seven boxes of commercial 38 caliber ammunition with the goals of associating cartridges within a box and matching residues to unfired powders, based on the P/S value and the qualitative identity of the additives. Gunpowder samples from four of the seven boxes of ammunition could be easily differentiated. When visual comparisons of the cartridge powders were considered in addition to composition, powder samples from all seven boxes of ammunition could be reliably differentiated. Handgun OGSR was also collected and evaluated in bulk as well as for individual particles. In some cases, residues could be reliably differentiated based on P/S and additive identity. It was instructive to evaluate the composition of individual unfired gunpowder and OGSR particles. We determined that both the numerical centroid and dispersity of the P/S measurements provide information for associations and exclusions. Associating measurements from residue particles with those of residue samples collected from a test firing of the same weapon and ammunition appears to be a useful approach to account for any changes in composition that occur during the firing process. PMID- 11908594 TI - A study of the effects of a Micelle Encapsulator Fire Suppression Agent on dynamic headspace analysis of fire debris samples. AB - The effects of a Micelle Encapsulator Fire Suppression Agent (F-500, Hazard Control Technologies Inc., Fayetteville, Georgia) on the routine analysis of fire debris samples by Gas Chromatography (GC) were studied. When mixed with water the product can be used in the suppression of Class A and Class B fires. Laboratory tests were performed to determine whether or not the product has any effect on the analysis for ignitable liquids by GC, in particular for gasoline, medium petroleum distillates. and heavy petroleum distillates. Test burns were suppressed using either the micelle encapsulator or water and samples collected from these burns were analyzed. The results of analysis show that use of the micelle encapsulator at a fire scene may affect the chromatographic data obtained from samples collected by the investigator. However, the effect does not prevent the identification of common ignitable liquids in fire debris samples. PMID- 11908595 TI - Classification of narcotics in solid mixtures using principal component analysis and Raman spectroscopy. AB - Eighty-five solid samples consisting of illegal narcotics diluted with several different materials were analyzed by near-infrared (785 nm excitation) Raman spectroscopy. Principal Component Analysis (PCA) was employed to classify the samples according to narcotic type. The best sample discrimination was obtained by using the first derivative of the Raman spectra. Furthermore, restricting the spectral variables for PCA to 2 or 3% of the original spectral data according to the most intense peaks in the Raman spectrum of the pure narcotic resulted in a rapid discrimination method for classifying samples according to narcotic type. This method allows for the easy discrimination between cocaine, heroin, and MDMA mixtures even when the Raman spectra are complex or very similar. This approach of restricting the spectral variables also decreases the computational time by a factor of 30 (compared to the complete spectrum), making the methodology attractive for rapid automatic classification and identification of suspect materials. PMID- 11908596 TI - Content based information retrieval in forensic image databases. AB - This paper gives an overview of the various available image databases and ways of searching these databases on image contents. The developments in research groups of searching in image databases is evaluated and compared with the forensic databases that exist. Forensic image databases of fingerprints, faces, shoeprints, handwriting, cartridge cases, drugs tablets, and tool marks are described. The developments in these fields appear to be valuable for forensic databases, especially that of the framework in MPEG-7, where the searching in image databases is standardized. In the future, the combination of the databases (also DNA-databases) and possibilities to combine these can result in stronger forensic evidence. PMID- 11908597 TI - Assessment of the quality of medical documents issued in central police stations in Madrid, Spain: the doctor's role in the prevention of ill-treatment. AB - Doctors sometimes assess allegations of ill-treatment. Reports from such examinations may be used if the practice of the police is to be appraised: they should therefore be relevant and exhaustive. We assessed, retrospectively, the quality of 318 medical documents concerning 100 persons held in central police stations in Madrid, Spain, from 1991 to 1994. In 71 documents concerning 44 persons the doctors quoted the detainee as alleging ill-treatment. Most of the documents appeared to lack significant information on history of ill-treatment and description of the clinical examination. Of 34 conclusions, ten were unacceptable and the premises were insufficient in 16. These observations point to weaknesses and needs for improvements in the fulfillment of the role of doctors as safeguards of the rights of detainees. Medical examinations should be conducted outside the control of police officers, by a neutral doctor using a check-list/protocol. The quality of the report should fulfill international standards. PMID- 11908598 TI - The effect of oleoresin capsicum "pepper" spray inhalation on respiratory function. AB - We performed a randomized, cross-over controlled trial to assess the effect of Oleoresin capsicum (OC) spray inhalation on respiratory function by itself and combined with restraint. Thirty-five subjects were exposed to OC or placebo spray, followed by 10 min of sitting or prone maximal restraint position (PMRP). Spirometry, oximetry, and end-tidal CO2 levels were collected at baseline and throughout the 10 min. Data were compared between groups (ANOVA) and with predefined normal values. In the sitting position, OC did not result in any significant changes in mean percent predicted forced vital capacity (%predFVC), percent predicted forced expiratory volume in 1 s (%predFEV1), oxygen, or CO2 levels. In PMRP, mean %predFVC and %predFEV1 fell 14.4 and 16.5% for placebo and 16.2 and 19.1% for OC, but were not significantly different by exposure. There was no evidence of hypoxemia or hypercapnia in either groups. OC exposure did not result in abnormal spirometry, hypoxemia, or hypoventilation when compared to placebo in either sitting or PMRP. PMID- 11908599 TI - Fiery tongues and mystical motivations: glossolalia in a forensic population is associated with mania and sexual/religious delusions. AB - Comparisons are made between a nonrandom sample of 18 glossolalists and 130 non glossolalists admitted to a maximum-security forensic hospital. The glossolalic mentally disordered offender exhibited a predominance of diagnoses in the manic spectrum, and was typically psychotic. The delusions, hallucinations, and crimes were predominately of a religious and sexual nature. Glossolalist perpetrators tended to be female. We review the extant research on glossolalia in both normal and clinical samples. and integrate our findings, the first study of glossolalia in a forensic setting. PMID- 11908600 TI - Double parricide: forensic analysis and psycholegal implications. AB - In a retrospective study, eleven adult parricidal forensic cases from Southern California are presented. Each case involves the murder of both parents and was referred for forensic evaluation. Common characteristics among the eleven cases are presented. Two case examples illustrate features of recognized adult parricidal subtypes. The findings are compared with studies involving parricide, double-parricide, and extant case law. PMID- 11908601 TI - Skeletal identification using the frontal sinus region: a retrospective study of 39 cases. AB - The importance of identification using the frontal sinus has been previously demonstrated in case reports. In this study, 39 cases of identification using frontal sinus comparison from the Ontario Chief Coroner's Office were reviewed and differences between antemortem and postmortem radiographs examined. All cases involved decedents older than twenty years. Three cases were rejected due to poor antemortem and postmortem film quality. One subject had no frontal sinus. Thirty five cases provided conclusive postmortem to antemortem pattern matches. Sixteen cases also yielded metric (quantitative) matches. Duration between antemortem and postmortem radiographic examinations, age, gender, and cause of death did not affect the ability to obtain a match. This is the largest study undertaken on actual cases and demonstrates the validity of frontal sinus pattern matching for forensic identification. PMID- 11908602 TI - Changes in composition of ballpoint pen inks on aging in darkness. AB - A method for comparison of the relative age of ink entries written by the same ballpoint pen on documents stored in darkness is presented. Inks were extracted from the document and analyzed by HPLC (high performance liquid chromatography). On aging, changes in the chemical composition of the inks were noted. These changes were similar to those observed when inks were exposed to light or heat. The aging was followed by using ternary diagrams constructed for dyes generally present in blue-colored inks--Crystal Violet, Methyl Violet, and Tetramethyl Para Rosaniline. The procedure is applicable for relative dating of ink entries in diaries, notebooks, etc., where often several ink entries are written by the same ink. However, a prolonged exposure of the document to daylight and/or artificial light (light from fluorescent tubes) as well as to extensive heat will render the whole procedure inapplicable. An example of the use of the proposed method in casework is given. PMID- 11908603 TI - Validation of a radiographic method to establish new fetal growth standards: radio-anatomical correlation. AB - In forensic medicine, specialists might face difficulties when estimating age at death from fetal remains. Depending on the state of preservation, this age assessment is essentially based on the diaphyseal size of long bones. In a previous work, for the measurement of fetal femoral ossified shafts, we already established a simple and reliable method using a radiographic protocol. Since we previously stated that radiographic measurement values were closer to real anatomical size than ultrasonographic ones, in the present study we decided to check the importance of the difference between radiographic and anatomical measurements. Therefore, we dissected 30 pairs of fetal femurs and compared the difference between the two kinds of measurements (in percentages). This difference seemed to be slight (4.027%), but it was large enough to entail significant differences (p < 0.001). In order to provide a correction factor for radiographic measurements, we established a linear regression formula, which was tested on another sample of 30 pairs of dissected femurs. As a consequence of the good results, we improved the linear regression using a powerful statistical tool: the bootstrap. Finally, we obtained a simple equation that allowed us to figure out the real anatomical size with an R2 of 99% and a mean relative difference of 0.153% (with a standard error of 0.252 mm, and therefore a 95% confidence interval with limits of -0.35 and 0.657 mm). This difference did not entail any significant differences (p = 0.498), and therefore, we concluded that with the proposed correction, radiographic measurements can easily be used by forensic specialists in their daily tasks or to establish new growth standards in order to best fit their population of interest. PMID- 11908604 TI - Extension of the color suite available for chemical enhancement of fingerprints in blood. AB - The use of ortho- and para-phenylenediamine (OPD & PPD respectively) for the enhancement of fingerprints in blood has been investigated. Optimal pH conditions and H2O2 concentrations have been determined using UV/Vis spectroscopy. Both OPD and PPD are effective and less hazardous alternatives to the presently used 3,3' diaminobenzidine (DAB) for the development of blood fingerprints, especially on porous surfaces. The orange color of OPD and the purple color of PPD offer alternative colors to the brown color of DAB and the light green color of ABTS for standing out against particular backgrounds. Both OPD and PPD can be used after ninhydrin treatment, but the reverse is not the case. PMID- 11908605 TI - Determination of ABO blood grouping from human oral squamous epithelium by the highly sensitive immunohistochemical staining method EnVision+. AB - Using the highly sensitive immunohistochemical staining method EnVision+, which employs a dextran polymer reagent for the secondary antibody, the detection of the ABH antigens was attempted in the oral squamous epithelium. This new technique uses monoclonal antibody as a primary antibody and it takes about three hours for staining. The time is much shorter than conventional absorption-elution testing or absorption-inhibition testing for the determination of ABO blood grouping. Secretor saliva samples were stained at strong intensity by the antibody, which corresponded to its blood group and anti-H. On the one hand, nonsecretor saliva samples were stained at strong intensity only by the antibody that corresponded to its blood group, and at weak intensity only by anti-H. Since human oral squamous epithelium antigens were stained specifically by this method, we can examine the ABO blood group of saliva samples and perform cytodiagnosis at the same time. Our research suggested that the EnVision+ Method is a useful technique for ABO blood grouping of saliva in forensic cases. PMID- 11908607 TI - Digital imaging methods as an aid in dental identification of human remains. AB - The physical comparison of known (K) and questioned (Q) evidence samples is an accepted tool in numerous forensic identification disciplines (1). A subset of this process is the use of antemortem and postmortem dental radiographs to identify unidentified human remains. This method has been generally accepted for decades (2). The outcome is performed with a considerable degree of accuracy, due in part to a finite pool of possible candidates for identification derived via the NCIC database, passenger lists, and law enforcement Missing Persons reports. This paper describes a dental identification comparison protocol that incorporated digital imaging technology in this process. The computer was used to create digital exemplars of the K and Q evidence that were spatially and quantitatively compared (3). The digital mode allowed direct metric and morphologic comparison through the aid of a digital camera, desktop computer, monitor, and printer. The well-known computer program Adobe Photoshop 5.0 (4) was used to process the digital information in two forensic cases described in this paper. It is a commercially available digital imaging editing program that is operated on laptop and desktop computers possessing sufficient chip speed and RAM (Pentium II or equivalent and at least 76MB RAM) to open the large-size files generated by high-resolution digital capture devices. This program accepts raster based image formats (e.g. .JPG, .BMP). Photoshop is noted for its diverse imaging functions, which allow the computer monitor to be used as a comparison microscope when Q and K sample images are tiled side-by-side and/or superimposed. Two and three-dimensional Q and K evidence samples can be individually digitized and then independently resized to allow two-dimensional comparison. The investigator also has the ability to create magnified images (200% to 300%) when the original digital image has been captured at near photoquality resolution (300 dpi). The visual comparison of physical features on the computer monitor permits a large field of view and robust digital control over image quality. Photographic measurement and enhancement features of Adobe Photoshop mimics and in some circumstances surpasses the historic use of conventional photographic manipulation in forensic casework. This paper presents two cases processed via routine forensic odontology identification protocols. These protocols had minimal results due to limitations described in the case histories. The additional application of digital methods proved useful in the ultimate identification of these human remains. PMID- 11908606 TI - Detection of a primer-binding site polymorphism for the STR locus D16S539 using the Powerplex 1.1 system and validation of a degenerate primer to correct for the polymorphism. AB - Quality assurance samples submitted from the NCSBI as part of a contract with TBTG to outsource DNA Database samples showed unexpected discrepancies for the locus D16S539 when all other loci yielded identical results. Discrepancies observed included allele drop out and an imbalance in sister alleles with samples returned from TBTG. This led to a comprehensive review of the technical procedures used between the two laboratories to determine the cause of the discrepancies noted for the locus D16S539, since both laboratories were using the PowerPlex 1.1 typing kit from the Promega Corporation. The NCSBI and the TBTG utilize different extraction methods (organic extraction vs. FTA) and amplification conditions (AmpliTaq vs AmpliTaq Gold), respectively, so the exact cause of discrepancy observed was not immediately apparent. Experiments at the NCSBI associated the observed allele drop out and the imbalance of the sister alleles with the use of AmpliTaq Gold and a hot start procedure. Sequencing data revealed that a point mutation resides on the D16S539 primer-binding site that reaches polymorphic levels in African-American populations. This led to the development of a degenerate primer by the Promega Corporation to detect "missing" alleles when AmpliTaq Gold is used. The degenerate primer was then thoroughly tested to show its efficacy in detecting the "true" D16S539 profile when used. PMID- 11908608 TI - Pseudo-subarachnoid hemorrhage of the head diagnosed by computerized axial tomography: a postmortem study of ten medical examiner cases. AB - In this report, we describe ten cases of pseudo-subarachnoid hemorrhage on computer axial tomography (CT) scan of the head. A pseudo-subarachnoid hemorrhage is a false positive finding by CT of the head in which the scan is interpreted as being positive for a subarachnoid hemorrhage not substantiated by subsequent neuropathologic findings. This study is a retrospective review of postmortem cases brought into the Office of the Chief Medical Examiner for the State of Maryland over a three-year period (from 1997 to 2000). We compared the clinician's impression of the CT scan with the postmortem neuropathology. The clinical diagnosis of subarachnoid hemorrhage was based on misinterpretation of non-contrast CT scans of the head. In six of the ten cases, the reading was performed by a radiologist and in four cases by nonradiologist physicians (emergency room physician, neurologist, or neurosurgeon). All the patients survived between a few hours to a few days after being admitted to the hospital. For most of the cases (80%), the neuropathology showed hypoxic/ischemic encephalopathy. The most common cause of death (four out of ten cases) was narcotic intoxication. This report is submitted so that clinicians and pathologist become more familiar with this entity. PMID- 11908609 TI - Concentrations of unconjugated morphine, codeine and 6-acetylmorphine in urine specimens from suspected drugged drivers. AB - Concentrations of unconjugated morphine, codeine and 6-acetylmorphine (6-AM), the specific metabolite of heroin, were determined in urine specimens from 339 individuals apprehended for driving under the influence of drugs (DUID) in Sweden. After an initial screening analysis by immunoassay for 5-classes of abused drugs (opiates, cannabinoids, amphetamine analogs, cocaine metabolite and benzodiazepines), all positive specimens were verified by more specific methods. Opiates and other illicit drugs were analyzed by isotope-dilution gas chromatography-mass spectrometry (GC-MS). The limits of quantitation for morphine, codeine and 6-AM in urine were 20 ng/mL. Calibration plots included an upper concentration limit of 1000 ng/mL for each opiate. We identified the heroin metabolite 6-AM in 212 urine specimens (62%) at concentrations ranging from 20 ng/mL to > 1000 ng/mL. The concentration of 6-AM exceeded 1000 ng/mL in 79 cases (37%) and 31 cases (15%) were between 20 and 99 ng/mL. When 6-AM was present in urine the concentration of morphine was above 1000 ng/mL in 196 cases (92%). The concentrations of codeine in these same urine specimens were more evenly distributed with 35% being above 1000 ng/mL and 21% below 100 ng/mL. These results give a clear picture of the concentrations of unconjugated morphine, codeine and 6-acetylmorphine that can be expected in opiate-positive urine specimens from individuals apprehended for DUID after taking heroin. PMID- 11908610 TI - Radiographic identification using the clavicle of an individual missing from the Vietnam conflict. AB - A case is reported in which radiographic comparison of a clavicle was used to establish the identification of a civilian missing from the Vietnam conflict. While the use of radiographic comparison of skeletal features is not a newly developed technique for personal identification purposes, this case outlines a unique set of circumstances surrounding the disappearance of an individual during the Vietnam conflict and his eventual identification. A radiographic comparison of a right clavicle was critical in the identification process of this individual almost 34 years after he was reported missing. The use of digital technology for radiographic comparison greatly facilitated the process. PMID- 11908611 TI - Restoration of obliterated painted registration number on vehicle. AB - A simple technique employed for the restoration of the obliterated painted registration number on a motor vehicle involved in a case of murder is described. PMID- 11908612 TI - An unusual oxidation type on bulb filament after a car crash dive. AB - A car with two dead bodies trapped inside was discovered in a gravel pit. The main hypothesis of investigators was a traffic accident simulation by night. Examination of a broken light-bulb revealed both a rather unusual oxidation type and small rounded cavities never reported before. Tests were performed and allowed forensic scientists to establish that the lightbulbs were switched on during the crash dive, in contrast with the investigators idea. PMID- 11908613 TI - Nutrient canals of the alveolar process as an anatomic feature for dental identifications. AB - Nutrient canals are anatomic structures of the alveolar bone through which neurovascular elements transit to supply teeth and supporting structures. A dental identification using a nutrient canal of the mandibular alveolar process as the most compelling anatomic feature for antemortem-postmortem radiographic comparison is described. Nutrient canals as a potential marker for clinical disease is also discussed. PMID- 11908614 TI - The spectrum of intramyocardial small vessel disease associated with sudden death. AB - Intramyocardial small vessel abnormalities are not commonly recognized. The best known abnormality is fibromuscular dysplasia involving the sinoatrial or atrioventricular nodal arteries. Small vessel disease has been reported as an isolated cardiac anomaly in individuals with sudden death, and may also be associated with other cardiac conditions including hypertrophic cardiomyopathy and mitral valve prolapse. The nature of the association is unknown, and the mechanism causing sudden death is sometimes obscure. We describe pathological changes of the intramyocardial small vessels of three individuals with sudden death. Abnormalities involved small vessels at different levels. In all the cases, the abnormalities were thought to have caused or contributed to the individual's death. The possible mechanisms of this are discussed. PMID- 11908615 TI - Bizarre impalement fatalities--where is the implement? AB - Two fatalities due to unusual impalement injuries are reported. (1) A large branch broken off during a storm had entered a passing car and perforated the chest of the driver and the back of the seat. The chest organs were grossly lacerated. The car was subsequently stopped by another tree and this second impact removed the wood from the body. (2) A man suffered anorectal impalement by the leg of a stool turned upside down. He had introduced one stool leg into his anus for sexual stimulation and fell onto it. This resulted in a wound channel 36 cm long including perforation of the rectum, urinary bladder, mesentery, transverse mesocolon and liver. Before autopsy, the mode of death was unclear because the man had removed the stool leg himself, his wife had hidden the stool from the scene, and there were no relevant external injuries. In both cases, a reliable reconstruction required investigation of the scene and consideration of extremely unlikely circumstances or of bizarre human activities. PMID- 11908616 TI - Cervical fracture, decapitation, and vehicle-assisted suicide. AB - Two cases of vehicle-assisted suicides are described in males aged 33 and 24 years, respectively. In both cases the victims had tied ropes between stationary objects and their necks and then attempted to drive their vehicles away. The speed with which the vehicles were driven resulted in forces great enough to cause fracture--dislocation of the cervical spine in Case 1 and virtual decapitation in Case 2. Although inadvertent alteration of the death scene in Case 1, with removal of the rope, complicated the initial assessment, the extent of soft tissue and bony injuries was such that ligature strangulation appeared unlikely. PMID- 11908617 TI - Spousal homicide and the subsequent staging of a sexual homicide at a distant location. AB - The case of a 63-year-old man who killed his 52-year-old wife and then staged a sexual homicide at a distant location is reported. A review of all evidence, a forensic psychological interview, and psychological testing indicated that the murder was the result of a narcissistic rage reaction during which the subject beat his wife to death with a paint can, a clothing iron, and a rock. He then drove her body to a field 87.3 miles away, and positioned it in a manner that exposed her breasts and her underwear. He turned himself into the police two days later. There is no controlled empirical research on staging, although this single case supports the criminal investigative theory that staging exists, and is done to deliberately mislead homicide investigations (Douglas et al., 1992). PMID- 11908618 TI - An unusual rubber stamp case. AB - Given a questioned document and a questioned pre-inked rubber stamp, comparisons of each can be made with a microscope or similar magnification device. If sufficient evidence exists, these questioned items can be linked even if the stamp was not used as it was originally designed. PMID- 11908619 TI - Postmortem blood and vitreous humor ethanol concentrations in a victim of a fatal motor vehicle crash. AB - A 20-year-old male was found on the passenger side of a small car after a collision with a semi-trailer truck. Postmortem blood, collected from the chest cavity, and vitreous humor samples were collected following harvesting of the heart and bones. Gas chromatographic analysis revealed a blood ethanol concentration of 0.32 g/dL and a vitreous humor ethanol concentration of 0.09 g/dL. The stomach was intact and full of fluid and food, but its contents were not collected. Possible explanations for the large difference between the two results include diffusion of ethanol from the stomach into the chest cavity, contamination of the blood sample prior to collection, and ingestion of a large quantity of ethanol shortly before death. This case demonstrates the importance of proper quality assurance procedures in collecting postmortem specimens and of collecting a vitreous humor sample for ethanol analysis in postmortem toxicology cases. PMID- 11908620 TI - Han Chinese population data for ten STR loci in Changsha, China. PMID- 11908621 TI - Allele frequencies for the AmpFlSTR profiler loci in a Colombian population (Department of Boyaca). PMID- 11908622 TI - Allele frequencies for eight STR loci in a population sample from Sicily (Italy). PMID- 11908623 TI - Allele frequency distribution at five Y-chromosomal short tandem repeat loci among five distinct ethnic groups of India. PMID- 11908624 TI - Indian population data for two tetrameric short tandem repeat loci--vWA and D3S1359. PMID- 11908625 TI - Distribution of allele frequencies for two short tandem repeats (HUMTH01 and F13A01) among five Indian population groups. PMID- 11908626 TI - Distribution of DYS19, DYS389 I, DYS389 II, DYS390 alleles in a southern Italian population sample. PMID- 11908627 TI - STR data for the PowerPlex 16 loci in Buenos Aires population (Argentina). PMID- 11908628 TI - Y-chromosome STR haplotypes in Central-West African immigrant in Spain population sample. PMID- 11908629 TI - Commentary on: McBride DG, Dietz MJ, Vennemeyer MT, Meadors SA, Benfer RA, Furbee L. Bootstrap methods for sex determination from the os coxae using the ID3 algorithm. PMID- 11908630 TI - Commentary on: Dou C, Bournique J, Zinda M, Gnezda M, Nally A, Salamone S. Comparison of rates of hydrolysis of lorazepam-glucuronide, oxazepam-glucuronide and temazepam-glucuronide catalyzed by E. coli beta-glucuronidase using the on line benzodiazepine screening immunoassay on the Roche/Hitachi 917 analyzer. PMID- 11908631 TI - Commentary on: Lee GSH, Brinch KM, Kannangara K, Dawson M, Wilson MA. A methodology based on NMR spectroscopy for the forensic analysis of condoms. PMID- 11908632 TI - The physiology of lactoferrin. AB - This paper reviews our current knowledge of the structure and function of the iron-binding protein lactoferrin. In particular, it attempts to relate the various proposed physiological functions of lactoferrin to its most characteristic biochemical properties, i.e. its ability to bind iron and its highly basic nature. The extent to which various physiological functions can be considered as definitely established is critically reviewed, and suggestions for future research are proposed. PMID- 11908633 TI - Bovine lactoferrin and lactoferricin derived from milk: production and applications. AB - Bovine lactoferrin is produced on an industrial scale from cheese whey or skim milk. The safety of purified lactoferrin has been confirmed from the results of a reverse mutation test using bacteria, a 13-week oral repeated-dose toxicity study in rats, and clinical studies. In order to apply active lactoferrin to various products, a process for its pasteurization was developed. Subsequently, lactoferrin has been used in a wide variety of products since it was first added to infant formula in 1986. A pepsin hydrolysate of lactoferrin is also used in infant formula. This hydrolysate contains a potent antimicrobial peptide named lactoferricin that is derived from the lactoferrin molecule by pepsin digestion. Semilarge-scale purification of lactoferricin can be performed by hydrophobic interaction chromatography. Lactoferricin also exhibits several biological actions and appears to be the functional domain of lactoferrin. Recent studies have demonstrated that oral administration of lactoferrin or lactoferricin exerts a host-protective effect in various animals and in humans. The results of these studies strongly suggest that the effects of oral lactoferrin are mediated by modulation of the immune system. Further elucidation of the clinical efficacy and mechanism of action of lactoferrin will increase the value of lactoferrin containing products. PMID- 11908634 TI - Effect of lactoferrin on Helicobacter felis induced gastritis. AB - Lactoferrin possesses antibiotic, antiinflammatory, and immune-modulating properties that may be active against the gastritis-, ulcer- and cancer-inducing bacterium Helicobacter pylori. In vitro testing of bovine and human lactoferrin by several laboratories has shown significant bacteriostatic and bactericidal activity. Subsequent in vivo testing of bovine lactoferrin in animal models of H. pylori infection has shown beneficial effects of this agent. Our laboratory has utilized a mouse model that is infected with the feline strain of this bacterium, H. felis. The resulting gastritis that develops in this model and the effects of bovine lactoferrin and recombinant human lactoferrin (from Aspergillus niger var. awamori, Agennix Inc., Houston, Tex.) treatment were assessed by various measures. Infected animals treated with orally administered lactoferrin showed reversals in all parameters. In addition, when recombinant human lactoferrin was used in combination with low doses of amoxicillin or tetracycline, there was an enhancement in gastritis-reducing activity. Possible mechanisms for these effects of lactoferrin are discussed. Lactoferrin has significant, orally active in vivo actions and should be further investigated for clinical situations involving Helicobacter infections where it may have utility when administered alone and also when given in combination with established antibiotic agents. PMID- 11908635 TI - Anti-invasive activity of bovine lactoferrin towards group A streptococci. AB - Group A streptococci (GAS) are able to invade cultured epithelial and endothelial cells without evidence of intracellular replication. GAS, like other facultative intracellular bacterial pathogens, evolved such ability to enter and to survive within host cells avoiding the host defences, and bacterial intracellular survival could explain the recurrence of infections. We report here that 1 mg bovine lactoferrin (bLf)/mL significantly hindered the in vitro invasion of cultured epithelial cells by GAS isolated from patients suffering from pharyngitis and completely inhibited the invasiveness of GAS pretreated with subinhibiting concentrations of erythromycin or ampicillin. One milligram of bLf per millilitre was also able to increase the number of epithelial cells undergoing apoptosis following GAS invasion, although the number of intracellular GAS in the presence of bLf decreased by about 10-fold. The ability of bLf to decrease GAS invasion was confirmed by an in vivo trial carried out on 12 children suffering from pharyngitis and already scheduled for tonsillectomy. In tonsil specimens from children treated for 15 days before tonsillectomy with both oral erythromycin (500 mg t.i.d. (three times daily)) and bLf gargles (100 mg t.i.d.), a lower number of intracellular GAS was found in comparison with that retrieved in tonsil specimens from children treated with erythromycin alone (500 mg t.i.d.). PMID- 11908636 TI - Antiviral activity of ovotransferrin discloses an evolutionary strategy for the defensive activities of lactoferrin. AB - Ovotransferrin (formerly conalbumin) is an iron-binding protein present in birds. It belongs to the transferrin family and shows about 50% sequence homology with mammalian serum transferrin and lactoferrin. This protein has been demonstrated to be capable of delivering iron to cells and of inhibiting bacterial multiplication. However, no antiviral activity has been reported for ovotransferrin, although the antiviral activity of human and bovine lactoferrins against several viruses, including human herpes simplex viruses, has been well established. In this report, the antiviral activity of ovotransferrin towards chicken embryo fibroblast infection by Marek's disease virus (MDV), an avian herpesvirus, was clearly demonstrated. Ovotransferrin was more effective than human and bovine lactoferrins in inhibiting MDV infection and no correlation between antiviral efficacy and iron saturation was found. The observations reported here are of interest from an evolutionary point of view since it is likely that the defensive properties of transferrins appeared early in evolution. In birds, the defensive properties of ovotransferrin remained joined to iron transport functions; in mammals, iron transport functions became peculiar to serum transferrin, and the defensive properties towards infections were optimised in lactoferrin. PMID- 11908637 TI - Cancer prevention by bovine lactoferrin and underlying mechanisms--a review of experimental and clinical studies. AB - In experimental studies, bovine lactoferrin (bLF) has been found to significantly inhibit colon, esophagus, lung, and bladder carcinogenesis in rats when administered orally in the post-initiation stage. Furthermore, concomitant administration with carcinogens resulted in inhibition of colon carcinogenesis, possibly by suppression of phase I enzymes, such as cytochrome P450 1A2 (CYP1A2), which is preferentially induced by carcinogenic heterocyclic amines. Enhancement of the activities of their phase II counterparts, such as glutathione S transferase might have also played a critical role in post-initiation suppression in a study of tongue carcinogenesis. Anti-metastatic effects were moreover detected when bLF was given intragastrically to mice bearing highly metastatic colon carcinoma 26 cells (Co 26Lu), with apparent enhancing influence on local and systemic immunity. Marked increase in the number of cytotoxic T and NK cells in the mucosal layer of the small intestine and peripheral blood cells was thus found, this in turn enhancing the production of Interleukin 18 (IL-18) and caspase-1 in the epithelial cells of the small intestine, with possible consequent induction of interferon (IFN)-gamma positive cells. Furthermore, bLF has been found to exert anti-hepatitis C virus (HCV) activity in a preliminary clinical trial in patients with chronic active hepatitis due to this virus, a main causative factor in hepatocellular carcinoma development in Japanese. More extensive clinical trials are now underway in the National Cancer Center Hospital and other institutes to further explore the preventive potential against colon carcinogenesis. PMID- 11908638 TI - Detection of exon polymorphisms in the human lactoferrin gene. AB - We previously demonstrated that lactoferrin gene polymorphisms occur in cancer cells of patients with leukemia and breast cancer. In this study, we established a non-radioactive polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, one of the most sensitive and simplest methods to detect polymorphisms and mutations of the human lactoferrin gene. We optimized the PCR conditions for nine different DNA templates and 16 pairs of exon primers for SSCP analysis. The DNA templates used in the analyses were prepared from a cosmid clone (CT6-1) that contains the human lactoferrin gene, human placental tissue, leukocytes from 10 normal volunteers, leukemic cells of two patients, and previously established three breast and two leukemic cell lines. With the appropriate exon-primer sets, PCR products from exon I to exon 16 of the lactoferrin gene were generated from the DNA templates and analyzed by SSCP. Compared with the homogenous cloned DNA, lactoferrin gene polymorphisms were detected within exons 2, 5, 7, 9, 13, 14, and 15 of the normal placental and leukocyte DNA. In addition, abnormal migration patterns of the lactoferrin gene in cancer cells were detected in exons 4, 5, 13, 14, and 15. The PCR-SSCP band migration patterns can be attributed either to gene polymorphism in normal cells or to DNA mutations in cancer cells and the employed method cannot distinguish between them. Nonetheless, the present analysis suggests that genetic polymorphisms of the lactoferrin gene exist in selected exons and additional mutations of the lactoferrin gene do occur in the cancer cells. PMID- 11908639 TI - Methoxychlor stimulates the mouse lactoferrin gene promoter through a GC-rich element. AB - The lactoferrin gene in the mouse uterus is a target gene for natural estrogens and xenoestrogens. One of the xenoestrogens is methyoxychlor, an insecticide that displays both estrogenic and antiandrogenic activities. Recently, methyoxychlor was found to stimulate lactoferrin gene expression in the uterus of an estrogen receptor null mouse. The present study is designed to uncover the methoxychlor response region in the mouse lactoferrin gene promoter. A series of different lengths of the mouse lactoferrin gene 5' flanking region were linked to a chloramphenicol acetyltransferase (CAT) reporter construct and transfected into human endometrial carcinoma HEC-1B cells, an estrogen receptor null cell line, in order to examine the methoxychlor response. The transfected cells were treated with methoxychlor or the metabolite of methoxychlor, HPTE, and the CAT reporter activities were measured. Constructs that contain a mouse lactoferrin 5' region longer than 100 bp were activated more than twofold by both methoxychlor and HPTE. The activation of the CAT reporter by the chemicals was dose dependent and reached saturation. Additional deletion mutants within the 100-bp region were tested, and a GC-rich sequence (GC-II) that we have previously characterized as an epidermal growth factor (EGF) response element was identified to be the region for the methoxychlor response. GC-II binds Sp1, Sp3, and IKLF transcription factors, collaborates with the AP1/CREB binding element, and confers the EGF response. Whether the effect of methoxychlor requires the AP1/CREB binding element has yet to be established; however, the present finding provides an alternative signaling pathway for the xenoestrogens. PMID- 11908640 TI - Lactoferrin and transferrin: functional variations on a common structural framework. AB - Lactoferrin shares many structural and functional features with serum transferrin, including an ability to bind iron very tightly, but reversibly, a highly-conserved three-dimensional structure, and essentially identical iron binding sites. Nevertheless, lactoferrin has some unique properties that differentiate it: an ability to retain iron to much lower pH, a positively charged surface, and other surface features that give it additional functions. Here, we review the structural basis for these similarities and differences, including the importance of dynamics and conformational change, and specific interactions that regulate iron binding and release. PMID- 11908641 TI - Studies of the ceruloplasmin-lactoferrin complex. AB - We have previously shown that iron-containing human lactoferrin (LF) purified from breast milk is able to form both in vitro and in vivo a complex with ceruloplasmin (CP), the copper-containing protein of human plasma. Here we present evidence that the CP-LF complex is dissociated by high concentrations of NaCl, CaCl2, or EDTA, or by decreasing the pH to 4.7. In addition, DNA, bacterial lipopolysaccharide, and heparin can displace CP from its complex with LF. Antibodies to either of the two proteins also cause dissociation of the complex. PMID- 11908642 TI - Ca2+ binding to bovine lactoferrin enhances protein stability and influences the release of bacterial lipopolysaccharide. AB - Bovine lactoferrin (bLf) is known to damage the outer membrane of Gram-negative bacteria by binding to bacterial lipopolysaccharide (LPS). We report that LPS is released from bacterial outer membranes also when apo- or metal-saturated Lf is separated from bacterial cells by a dialysis membrane. This process occurs in phosphate-buffered saline with no added Ca2+ and Mg2+ and is hindered by addition of these cations. The effect of bLf is similar to that induced by EDTA and has been ascribed to chelation of Ca2+. In fact, it may be envisaged that Ca2+ binding sites on LPS have different affinities and that bLf can remove those ions that are more weakly bound. Ca2+ binding does not alter Lf iron-binding properties significantly or its UV and CD spectral features but brings about changes in the FT-IR bands due to carboxylate residues. Ca2+ binding is characterized by an apparent dissociation constant of 6 microM and a stoichiometry of 1.55 Ca2+ per Lf molecule; it enhances bLf stability towards chemical and thermal denaturation. The increase in stability takes place in both the apo- and iron-saturated forms but not in the desialilated protein, indicating that the carboxylate groups of the sialic acid residues present on two of the glycan chains are involved in Ca2+ binding. PMID- 11908643 TI - Towards a structure-function analysis of bovine lactoferricin and related tryptophan- and arginine-containing peptides. AB - The iron-binding protein lactoferrin is a multifunctional protein that has antibacterial, antifungal, antiviral, antitumour, anti-inflammatory, and immunoregulatory properties. All of these additional properties appear to be related to its highly basic N-terminal region. This part of the protein can be released in the stomach by pepsin cleavage at acid pH. The 25-residue antimicrobial peptide that is released is called lactoferricin. In this work, we review our knowledge about the structure of the peptide and attempt to relate this to its many functions. Microcalorimetry and fluorescence spectroscopy data regarding the interaction of the peptide with model membranes show that binding to net negatively charged bacterial and cancer cell membranes is preferred over neutral eukaryotic membranes. Binding of the peptide destabilizes the regular membrane bilayer structure. Residues that are of particular importance for the activity of lactoferricin are tryptophan and arginine. These two amino acids are also prevalent in "penetratins", which are regions of proteins or synthetic peptides that can spontaneously cross membranes and in short hexapeptide antimicrobial peptides derived through combinatorial chemistry. While the antimicrobial, antifungal, antitumour, and antiviral properties of lactoferricin can be related to the Trp/Arg-rich portion of the peptide, we suggest that the anti-inflammatory and immunomodulating properties are more related to a positively charged region of the molecule, which, like the alpha- and beta defensins, may act as a chemokine. Few small peptides are involved in as wide a range of host defense functions as bovine and human lactoferricin. PMID- 11908644 TI - Important structural features of 15-residue lactoferricin derivatives and methods for improvement of antimicrobial activity. AB - This review focuses on important structural features affecting the antimicrobial activity of 15-residue derivatives of lactoferricins. Our investigations are based on an alanine-scan of a 15-residue bovine lactoferricin derivative that revealed the absolute necessity of two tryptophan residues for antimicrobial activity. This "tryptophan-effect" was further explored in homologous derivatives of human, caprine, and porcine lactoferricins by the incorporation of one additional tryptophan residue, and by increasing the content of tryptophan in the bovine derivative to five residues. Most of the resulting peptides display a substantial increase in antimicrobial activity. To identify which molecular properties make tryptophan so effective, a series of bovine lactoferricin derivatives were prepared containing non-encoded unnatural aromatic amino acids, which represented various aspects of the physicochemical nature of tryptophan. The results clearly demonstrate that tryptophan is not unique since most of the modified peptides were of higher antimicrobial potency than the native peptide. The size and three-dimensional shape of the inserted "super-tryptophans" are the most important determinants for the high antimicrobial activity of the modified peptides. This review also describes the use of a "soft-modeling" approach in order to identify important structural parameters affecting the antimicrobial activity of modified 15-residue murine lactoferricin derivatives. This QSAR-study revealed that the net charge, charge asymmetry, and micelle affinity of the peptides were the most important structural parameters affecting their antimicrobial activity. PMID- 11908645 TI - Lactoferrin gene expression and regulation: an overview. AB - Lactoferrin is highly conserved among human, mouse, bovine, and porcine species. The numbers of amino acids encoded by 15 of the 17 exons in these species are identical, and in 12 locations, they have identical codon interruptions at the intron-exon splice junctions. However, lactoferrin expression is both ubiquitous and species, tissue, and cell-type specific. It is differentially regulated through multiple signaling pathways such as steroid hormone, growth factor, and kinase cascade pathways. Comparing the lactoferrin gene promoters from different species, common and different characteristics are observed. The human, mouse, bovine, porcine, and bubaline (African antelope) promoters all contain a noncanonical TATA box with an adjacent Sp1 site. Both human and mouse have multiple steroid hormone response elements, while none are found in the other species studied, suggesting that the lactoferrin gene is differentially regulated among different species by steroid hormones. Several transcription factors have been identified that are crucial for the expression of the lactoferrin gene during differentiation of the myeloid cells and in estrogen and epidermal growth factor regulation. This article provides an overview on lactoferrin expression and regulation in different species. PMID- 11908646 TI - Characterization of mammalian receptors for lactoferrin. AB - Lactoferrin (Lf) has been suggested to have several physiological functions. Specific binding of Lf, indicating the presence of Lf receptors (LfRs), has been observed in various types of mammalian cells such as lymphocytes, hepatocytes, and enterocytes. These LfRs are considered to function as a mediator for some of the functions of Lf. We here review current knowledge of mammalian LfRs characterized in different tissues. We also briefly present evidence for the existence of an LfR provided by our cloning of a human intestinal LfR (HLfR). The entire coding region of the HLfR was cloned by polymerase chain reaction (PCR), and a recombinant HLfR (rHLfR) was expressed in a baculovirus system. The rHLfR was purified by immobilized human Lf (HLf) affinity chromatography, indicating that the rHLfR retained the capacity to bind HLf. The gene was expressed at high levels in fetal small intestine and in adult heart but at lower levels in Caco-2 cells. In summary, we demonstrate the presence of a unique receptor-mediated mechanism for Lf, functioning in the small intestine of the newborn infant and possibly in other tissues of human adults. PMID- 11908647 TI - Bacterial lactoferrin receptors: insights from characterizing the Moraxella bovis receptors. AB - Moraxella bovis is the causative agent of infectious conjunctivitis in cattle. Moraxella bovis isolates were shown to specifically bind bovine lactoferrin (bLf) and bovine transferrin (bTf) and to use these proteins as a source of iron to support the growth of iron-limited cells. Affinity isolation experiments with immobilized bTf yielded two proteins readily resolved by SDS-PAGE analysis, whereas only a single band of approximately 100 kDa was detected when immobilized bLf was used as the affinity ligand. Using a novel cloning strategy, regions containing the genes encoding the lactoferrin (Lf) and transferrin (Tf) receptor proteins were isolated and sequenced, demonstrating that they both consisted of two genes, with the tbpB or lbpB gene preceding the tbpA or lbpA gene. The cloned lbp genes were used to generate isogenic mutants deficient in lactoferrin binding protein A and (or) B, and the resulting strains were tested in growth and binding assays. The isogenic mutants were deficient in their use of bLf for growth and had substantially diminished bLf binding capability. The predicted amino acid sequence from the segment encoding Lf binding protein B revealed an internal amino acid homology suggesting it is a bi-lobed protein, with a C-lobe enriched in acidic amino acids, but without the evident clustering observed in Lf-binding proteins from other species. PMID- 11908648 TI - Lactoferrin-binding proteins in Bifidobacterium bifidum. AB - Lactoferrin is an iron-binding glycoprotein and its bacteriostatic and bactericidal effects on gram-positive and gram-negative bacteria are well known. On the other hand, it is known that certain kinds of lactic acid bacteria are resistant to its antibacterial effects. Moreover, it is reported that lactoferrin promotes the growth of bifidobacteria in in vitro and in vivo experiments. In our experiments, lactoferrin-binding protein was found both in the membrane and cytosolic fractions of Bifidobacterium bifidum Bb-11. The bifidobacteria were grown in anaerobic conditions with lactobacilli MRS broth containing cysteine, harvested by centrifugation, and processed by sonication. The lactoferrin-binding proteins on the PVDF-membrane transferred after SDS-PAGE were detected by far Western (western-Western) method using biotinylated lactoferrin and streptavidin labelled horse radish peroxidase. The molecular weights of the lactoferrin binding protein detected in the membrane fraction were estimated to be 69 kDa and those in cytosolic fractions were 20, 35, 50, and 66 kDa. PMID- 11908649 TI - Lactoferrin and host defense. AB - Lactoferrin is a multifunctional member of the transferrin family of nonheme iron binding glycoproteins. Lactoferrin is found at the mucosal surface where it functions as a prominent component of the first line of host defense against infection and inflammation. The protein is also an abundant component of the specific granules of neutrophils and can be released into the serum upon neutrophil degranulation. While the iron-binding properties were originally believed to be solely responsible for the host defense properties ascribed to lactoferrin, it is now known that other mechanisms contribute to the broad spectrum anti-infective and anti-inflammatory roles of this protein. In this article, current information on the functions and mechanism of action of lactoferrin are reviewed, with particular emphasis on the activities that contribute to this protein's role in host defense. In addition, studies demonstrating that lactoferrin inhibits allergen-induced skin inflammation in both mice and humans, most likely secondary to TNF-alpha (tumor necrosis factor alpha) production, are summarized. Collectively, these results suggest that lactoferrin functions as a key component of mammalian host defense at the mucosal surface. PMID- 11908650 TI - RAPD markers as predictors of infectious hypodermal and hematopoietic necrosis virus (IHHNV) resistance in shrimp (Litopenaeus stylirostris). AB - Random amplified polymorphic DNA (RAPD) fingerprints of two shrimp populations (Litopenaeus stylirostris) were compared to find genetic marker(s) that may be associated with infectious hypodermal and hematopoietic necrosis virus (IHHNV) resistance or susceptibility. Of the 100 10-mer random primers and 100 intersimple-sequence repeat (ISSR) primers screened, five provided markers specific to the Super Shrimp population and three provided markers specific to the wild caught population. The two populations were further characterized for relative viral load (reported as cycle threshold, CT) using real-time quantitative PCR with primers specific to the IHHNV genome. The beta-actin gene was amplified to serve as a control for normalization of the IHHNV viral load. The mean viral load was significantly lower (C(T) = 34.58; equivalent to 3.3 x 10(1) copies of IHHNV genome/ng DNA) in Super Shrimp than in the wild caught population (CT = 23.49; equivalent to 4.2 x 10(4) copies/ng DNA; P < 0.001; CT values are inversely related to viral load). A preliminary prediction model was created with Classification and Regression Tree (CART) software (Salford Systems, San Diego, Calif.), where the resultant decision tree uses the presence or absence of seven RAPD markers as predictors of the relative viral load. PMID- 11908651 TI - Genetic and physical mapping of Lrk10-like receptor kinase sequences in hexaploid oat (Avena sativa L.). AB - Oat receptor-like kinase gene sequences, homologous to the Lrk10 gene from wheat (Triticum aestivum L.), were mapped in oat (Avena sativa L.). PCR primers designed from the wheat Lrk10 were used to produce ALrk10 from oat. Two DNA sequences, ALrk1A1 and ALrk4A5, were produced from primers designed from coding and noncoding regions of ALrk10. Their use as RFLP probes indicated that the kinase genes mapped to four loci on different hexaploid oat 'Kanota' x 'Ogle' linkage groups (4_12, 5, 6, and 13) and to a fifth locus unlinked to other markers. Three of these linkage groups contain a region homologous to the short arm of chromosome I of wheat and the fourth contains a region homologous to chromosome 3 of wheat. Analysis with several nullisomics of oat indicated that two of the map locations are on satellite chromosomes. RFLP mapping in a 'Dumont' x 'OT328' population indicated that one map location is closely linked to Pg9, a resistance gene to oat stem rust (Puccinia graminis subsp. avenae). Comparative mapping indicates this to be the region of a presumed cluster of crown rust (Puccinia coronata subsp. avenae) and stem rust resistance genes (Pg3, Pg9, Pc44, Pc46, Pc50, Pc68, Pc95, and PcX). The map position of several RGAs located on KO6 and KO3_38 with respect to Lrk10 and storage protein genes are also reported. PMID- 11908652 TI - Cytogenetic and molecular characterization of intergeneric hybrids between Brassica napus and Orychophragmus violaceus. AB - Twenty-two intergeneric hybrids from a cross between Brassica napus (AACC, 2n = 38) cultivar Oro and the ornamental crucifer Orychophragmus violaceus (OO, 2n = 24) were produced without embryo rescue. The plants were classified into three groups based on morphological and cytological observations and RAPD banding patterns. Plants of Group I had morphological traits of both parents and 2n = 29 chromosomes. In these plants, 62.1% of the pollen mother cells (PMCs) had the pairing configuration 1 III + 9 II + 8 I; the remaining PMCs had 10 II + 9 I. The plants possessed 97.6-98.8% B. napus specific and 9.2-11.7% O. violaceus specific RAPD fragments. Plants of Group II exhibited novel morphological traits and possessed 2n = 35, 36, or 37 chromosomes. Plants of Group III were morphologically similar to B. napus and possessed 2n = 19, 37, 38, or 39 chromosomes. Plants of Group II and Group III had 94.1-99.4% B. napus specific RAPD fragments and no O. violaceus specific RAPD fragments. Chromosome fragments were observed in PMCs of most of the F1 plants in all groups. Based on the cytological results and RAPD analysis, it is suggested that genome doubling and chromosome elimination occurred in the intergeneric hybrids of B. napus x O. violaceus. PMID- 11908653 TI - A DNA mismatch repair gene links to the Ph2 locus in wheat. AB - DNA mismatch repair is an essential system for maintaining genetic stability in bacteria and higher eukaryotes. Based on the conserved regions of the bacterial MutS gene and its homologues in yeast and human, a wheat cDNA homologue of MSH6, designated TaMSH7, was isolated by RT-PCR. The deduced amino acid sequence of TaMSH7 shows conserved domains characteristic of other MSH6 genes, with highest similarity to maize MSH7 and Arabidopsis MSH7. TaMSH7 is expressed in meristem tissue associated with a high level of mitotic and meiotic activity, with maximum expression in the reproductive organs of young flower spikes. TaMSH7 is located on the short arms of chromosomes 3A, 3B, and 3D and has been mapped within barley chromosome 3HS. The copy on 3DS is located within the region deleted in the wheat mutant ph2a, which shows altered recombination frequency in the interspecific hybrids. The relationship between the ph2a mutant and TaMSH7 gene function is discussed. PMID- 11908654 TI - Allelic variation of a Beauveria bassiana (Ascomycota: Hypocreales) minisatellite is independent of host range and geographic origin. AB - The minisatellite locus, BbMin1, was isolated from a partial Beauveria bassiana genomic library that consisted of poly(GA) flanked inserts. Polymerase chain reaction (PCR) of the BbMin1 repeat demonstrated allele size variation among 95 B. bassiana isolates. Amplification was also observed from single isolates of Beauveria amorpha, Beauveria brongniartii, and Beauveria caledonica. Eight alleles were identified at the haploid locus, where repeat number fluctuated between one and fourteen. AMOVA and theta (Fst) indicated that fixation of repeat number has not occurred within pathogenic ecotypes or geographically isolated samples of B. bassiana. Selective neutrality of allele size, the rate of BbMin1 mutation, and the age of the species may contribute to host and geographic independence of the marker. Presence of alleles with a large number of repeat units may be attributed to the rare occurrence of somatic recombination or DNA replication error. The molecular genetic marker was useful for the identification of genetic types of B. bassiana and related species. PMID- 11908655 TI - Chromosomal and allelic variation in Drosophila americana: selective maintenance of a chromosomal cline. AB - Geographically structured genetic variation, as represented by clines and hybrid zones, offers unique opportunities to study adaptation and speciation in natural populations. A hybrid zone has been reported between Drosophila americana americana and Drosophila americana texana, two taxa that are distinguished solely by the arrangement of their X and 4th chromosomes. In this study, samples of D. americana were collected along a latitudinal transect across the inferred hybrid zone, and the frequency of the alternative chromosomal arrangements is reported. These data illustrate that the alternative chromosomal arrangements are distributed along a shallow cline over a broad geographic region, and that the frequency of the arrangements is tightly correlated with latitude. Allelic variants at 13 RFLP loci in three genes on chromosome 4 exhibit no evidence of association with the cline. Presence of a cline for the chromosomal arrangements, as well as a general absence of geographic structure for variation at these genes, is interpreted as evidence that natural selection is responsible for the maintenance of this chromosomal cline. Furthermore, these results demonstrate that taxonomic subdivision of D. americana is unwarranted, because it exists as a cohesive species that is segregating a chromosomal fusion. PMID- 11908656 TI - A molecular linkage map of tomato displaying chromosomal locations of resistance gene analogs based on a Lycopersicon esculentum x Lycopersicon hirsutum cross. AB - A molecular linkage map of tomato was constructed based on a BC1 population (N = 145) of a cross between Lycopersicon esculentum Mill. line NC84173 (maternal and recurrent parent) and Lycopersicon hirsutum Humb. and Bonpl. accession PI126445. NC84173 is an advanced breeding line that is resistant to several tomato diseases, not including early blight (EB) and late blight (LB). PI126445 is a self-incompatible accession that is resistant to many tomato diseases, including EB and LB. The map included 142 restriction fragment length polymorphism (RFLP) markers and 29 resistance gene analogs (RGAs). RGA loci were identified by PCR amplification of genomic DNA from the BC1 population, using ten pairs of degenerate oligonucleotide primers designed based on conserved leucine-rich repeat (LRR), nucleotide binding site (NBS), and serine (threonine) protein kinase (PtoKin) domains of known resistance genes (R genes). The PCR-amplified DNAs were separated by denaturing polyacrylamide gel electrophoresis (PAGE), which allowed separation of heterogeneous products and identification and mapping of individual RGA loci. The map spanned 1469 cM of the 12 tomato chromosomes with an average marker distance of 8.6 cM. The RGA loci were mapped to 9 of the 12 tomato chromosomes. Locations of some RGAs coincided with locations of several known tomato R genes or quantitative resistance loci (QRLs), including Cf-1, Cf 4, Cf-9, Cf-ECP2, rx-1, and Cm1.1 (chromosome 1); Tm-1 (chromosome 2); Asc (chrromosme 3); Pto, Fen, and Prf (chromosome 5); 01-1, Mi, Ty-1, Cm6.1, Cf-2, CF 5, Bw-5, and Bw-1 (chromosome 6); I-1, 1-3, and Ph-1 (chromosome 7); Tm-2a and Fr1 (chromosome 9); and Lv (chromosome 12). These co-localizations indicate that the RGA loci were either linked to or part of the known R genes. Furthermore, similar to that for many R gene families, several RGA loci were found in clusters, suggesting their potential evolutionary relationship with R genes. Comparisons of the present map with other molecular linkage maps of tomato, including the high density L. esculentum x Lycopersicon pennellii map, indicated that the lengths of the maps and linear order of RFLP markers were in good agreement, though certain chromosomal regions were less consistent than others in terms of the frequency of recombination. The present map provides a basis for identification and mapping of genes and QTLs for disease resistance and other desirable traits in PI126445 and other L. hirsutum accessions, and will be useful for marker-assisted selection and map-based gene cloning in tomato. PMID- 11908657 TI - Comparative mapping of Homo sapiens chromosome 4 (HSA4) and Sus scrofa chromosome 8 (SSC8) using orthologous genes representing different cytogenetic bands as landmarks. AB - The recently published draft sequence of the human genome will provide a basic reference for the comparative mapping of genomes among mammals. In this study, we selected 214 genes with complete coding sequences on Homo sapiens chromosome 4 (HSA4) to search for orthologs and expressed sequence tag (EST) sequences in eight other mammalian species (cattle, pig, sheep, goat, horse, dog, cat, and rabbit). In particular, 46 of these genes were used as landmarks for comparative mapping of HSA4 and Sus scrofa chromosome 8 (SSC8); most of HSA4 is homologous to SSC8, which is of particular interest because of its association with genes affecting the reproductive performance of pigs. As a reference framework, the 46 genes were selected to represent different cytogenetic bands on HSA4. Polymerase chain reaction (PCR) products amplified from pig DNA were directly sequenced and their orthologous status was confirmed by a BLAST search. These 46 genes, plus 11 microsatellite markers for SSC8, were typed against DNA from a pig-mouse radiation hybrid (RH) panel with 110 lines. RHMAP analysis assigned these 57 markers to 3 linkage groups in the porcine genome, 52 to SSC8, 4 to SSC15, and 1 to SSC17. By comparing the order and orientation of orthologous landmark genes on the porcine RH maps with those on the human sequence map, HSA4 was recognized as being split into nine conserved segments with respect to the porcine genome, seven with SSC8, one with SSC15, and one with SSC17. With 41 orthologous gene loci mapped, this report provides the largest functional gene map of SSC8, with 30 of these loci representing new single-gene assignments to SSC8. PMID- 11908658 TI - Identification and chromosomal organization of two rye genome-specific RAPD products useful as introgression markers in wheat. AB - Two rye genome-specific random amplified polymorphic DNA (RAPD) markers were identified for detection of rye introgression in wheat. Both markers were amplified in all of the tested materials that contained rye chromatin such as rye, hexaploid triticale, wheat-rye addition lines, and wheat varieties with 1BL.1RS translocation. Two cloned markers, designated pSc10C and pSc20H, were 1012 bp and 1494 bp, respectively. Sequence analysis showed that both pSc10C and pSc20H fragments were related to retrotransposons, ubiquitously distributed in plant genomes. Using fluorescence in situ hybridization (FISH), probe pSc10C was shown to hybridize predominantly to the pericentromeric regions of all rye chromosomes, whereas probe pSc20H was dispersed throughout the rye genome except at telomeric regions and nucleolar organizing regions. The FISH patterns showed that the two markers should be useful to select or track all wheat-rye translocation lines derived from the whole arms of rye chromosomes, as well as to characterize the positions of the translocation breakpoints generated in the proximal and distal regions of rye arms. PMID- 11908659 TI - A microsatellite sequence from the rice blast fungus (Magnaporthe grisea) distinguishes between the centromeres of Hordeum vulgare and H. bulbosum in hybrid plants. AB - A TC/AG-repeat microsatellite sequence derived from the rice blast fungus (Magnaporthe grisea) hybridized to all of the centromeres of Hordeum vulgare chromosomes, but hybridized faintly or not at all to the chromosomes of Hordeum bulbosum. Using this H. vulgare centromere-specific probe, the chromosomes of four F1 hybrids between H. vulgare and H. bulbosum were analyzed. The chromosome constitution in the root tips of the hybrids was mosaic, i.e., 7 (7v, H. vulgare) and 14 (7v + 7b H. bulbosum), or 14 (7v + 7b) and 27 (14v + 13b), or 7 (7v), 14 (7v + 7b), and 27 (14v + 13b). The 27-chromosome tetraploid hybrid cells were revealed to have the NOR (nucleolus organizer region) bearing chromosome of H. bulbosum in a hemizygous state, which might indicate some role for this chromosome in the chromosome instability of the hybrid condition. The chromosomal distribution showed that the chromosomes of H. vulgare were concentric and chromosomes of H. bulbosum were peripheral in the mitotic squash. This non-random chromosome distribution and the centromere-specific repeated DNA differences in the two species were discussed in relation to H. bulbosum chromosome elimination. Meiotic chromosome analyses revealed a high frequency of homoeologous chromosome pairing in early prophase. However, this chromosome pairing did not persist until later meiotic stages and many univalents and chromosome fragments resulted. These were revealed to be H. bulbosum by fluorescence in situ hybridization (FISH) analysis with the H. vulgare centromere-specific probe. Because the chromosome segregation of H. vulgare and H. bulbosum chromosomes at anaphase I of meiosis was random, the possibility for obtaining chromosome substitution lines in diploid barley from the diploid hybrid was discussed. PMID- 11908660 TI - An improved genetic linkage map for cowpea (Vigna unguiculata L.) combining AFLP, RFLP, RAPD, biochemical markers, and biological resistance traits. AB - An improved genetic linkage map has been constructed for cowpea (Vigna unguiculata L. Walp.) based on the segregation of various molecular markers and biological resistance traits in a population of 94 recombinant inbred lines (RILs) derived from the cross between 'IT84S-2049' and '524B'. A set of 242 molecular markers, mostly amplified fragment length polymorphism (AFLP), linked to 17 biological resistance traits, resistance genes, and resistance gene analogs (RGAs) were scored for segregation within the parental and recombinant inbred lines. These data were used in conjunction with the 181 random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), AFLP, and biochemical markers previously mapped to construct an integrated linkage map for cowpea. The new genetic map of cowpea consists of 11 linkage groups (LGs) spanning a total of 2670 cM, with an average distance of 6.43 cM between markers. Astonishingly, a large, contiguous portion of LG1 that had been undetected in previous mapping work was discovered. This region, spanning about 580 cM, is composed entirely of AFLP markers (54 in total). In addition to the construction of a new map, molecular markers associated with various biological resistance and (or) tolerance traits, resistance genes, and RGAs were also placed on the map, including markers for resistance to Striga gesnerioides races 1 and 3, CPMV, CPSMV, B1CMV, SBMV, Fusarium wilt, and root-knot nematodes. These markers will be useful for the development of tools for marker-assisted selection in cowpea breeding, as well as for subsequent map-based cloning of the various resistance genes. PMID- 11908661 TI - FISH-mapping of rDNAs and Arabidopsis BACs on pachytene complements of selected Brassicas. AB - To improve resolution of physical mapping on Brassica chromosomes, we have chosen the pachytene stage of meiosis where incompletely condensed bivalents are much longer than their counterparts at mitotic metaphase. Mapping with 5S and 45S rDNA sequences demonstrated the advantage of pachytene chromosomes in efficient physical mapping and confirmed the presence of a novel 5S rDNA locus in Brassica oleracea, initially identified by genetic mapping using restriction fragment length polymorphism (RFLP). Fluorescence in situ hybridization (FISH) analysis visualized the presence of the third 5S rDNA locus on the long arm of chromosome C2 and confirmed the earlier reports of two 45S rDNA loci in the B. oleracea genome. FISH mapping of low-copy sequences from the Arabidopsis thaliana bacterial artificial chromosome (BAC) clones on the B. oleracea chromosomes confirmed the expectation of efficient and precise physical mapping of meiotic bivalents based on data available from A. thaliana and indicated conserved organization of these two BAC sequences on two B. oleracea chromosomes. Based on the heterologous in situ hybridization with BACs and their mapping applied to long pachytene bivalents, a new approach in comparative analysis of Brassica and A. thaliana genomes is discussed. PMID- 11908662 TI - Rapid verification of wheat-Hordeum introgressions by direct staining of SCAR, STS, and SSR amplicons. AB - A range of single tagged site (STS), simple sequence repeat (SSR), and sequence characterized amplified region (SCAR) markers were screened for their utility in detecting Hordeum vulgare and H. chilense chromosomes in a wheat background. PCR conditions were optimized for specific amplification of the targeted sequences and to avoid cross-species amplification. Two H. vulgare derived STSs, six H. vulgare derived SSRs, and nine H. chilense derived SCARs were usable for the detection of five H. vulgare and three H. chilense chromosomes by direct ethidium bromide staining of the PCR products in test tubes, avoiding the more costly and time-consuming DNA electrophoresis step. The practical application of the method is illustrated by the identification of a monotelosomic substitution of H. vulgare chromosome 6HS in tritordeum and a monosomic addition of H. chilense chromosome 6Hch in durum wheat. PMID- 11908663 TI - Novel genes are enriched in normalized cDNA libraries from drought-stressed seedlings of rice (Oryza sativa L. subsp. indica cv. Nagina 22). AB - We have utilized an efficient method to enrich cDNA libraries for novel genes and genes responsive to drought stress in rice (Oryza sativa L. subsp. indica). We separately constructed standard and normalized cDNA libraries from leaf tissue of rice seedlings grown under controlled drought stress. Sequencing from the 3' end was performed on 1000 clones from the normalized leaf cDNA library and 200 clones from the standard leaf cDNA library. For the first 200 clones, the clone redundancy in the non-normalized library was about 10%, compared with 3.5% in the normalized cDNA library. Comparison of these cDNAs with the sequences in public databases revealed that 28.2% of the expressed sequence tags (ESTs) from the normalized library were novel. Clones from the standard and normalized leaf libraries and a root library uncovered numerous cDNAs that are highly homologous to known drought-responsive genes including those that encode metallothioneins, late embroyonic abundant (LEA) proteins, heat-shock proteins, cytochrome P450 enzymes, catalases, peroxidases, kinases, phosphatases, and transcription factors. PMID- 11908664 TI - Linkage mapping of genes controlling resistance to white rust (Albugo candida) in Brassica rapa (syn. campestris) and comparative mapping to Brassica napus and Arabidopsis thaliana. AB - Genes for resistance to white rust (Albugo candida) in oilseed Brassica rapa were mapped using a recombinant inbred (RI) population and a genetic linkage map consisting of 144 restriction fragment length polymorphism (RFLP) markers and 3 phenotypic markers. Young seedlings were evaluated by inoculating cotyledons with A. candida race 2 (AC2) and race 7 (AC7) and scoring the interaction phenotype (IP) on a 0-9 scale. The IP of each line was nearly identical for the two races and the population showed bimodal distributions, suggesting that a single major gene (or tightly linked genes) controlled resistance to the two races. The IP scores were converted to categorical resistant and susceptible scores, and these data were used to map a single Mendelian gene controlling resistance to both races on linkage group 4 where resistance to race 2 had been mapped previously. A quantitative trait loci (QTL) mapping approach using the IP scores detected the same major resistance locus for both races, plus a second minor QTL effect for AC2 on linkage group 2. These results indicate that either a dominant allele at a single locus (Acal) or two tightly linked loci control seedling resistance to both races of white rust in the biennial turnip rape cultivar Per. The map positions of white rust resistance genes in B. rapa and Brassica napus were compared and the results indicate where additional loci that have not been mapped may be located. Alignment of these maps to the physical map of the Arabidopsis genome identified regions to target for comparative fine mapping using this model organism. PMID- 11908665 TI - Mapping epigenetic quantitative trait loci (QTL) altering a developmental trajectory. AB - Genetic variation in a quantitative trait that changes with age is important to both evolutionary biologists and breeders. A traditional analysis of the dynamics of genetic variation is based on the genetic variance-covariance matrix among different ages estimated from a quantitative genetic model. Such an analysis, however, cannot reveal the mechanistic basis of the genetic variation for a growth trait during ontogeny. Age-specific genetic variance at time t conditional on the causal genetic effect at time t - 1 implies the generation of episodes of new genetic variation arising during the interval t - 1 to t. In the present paper, the conditional genetic variance estimated from Zhu's (1995) conditional model was partitioned into its underlying individual quantitative trait loci (QTL) using molecular markers in an F2 progeny of poplars (Populus trichocarpa and Populus deltoides). These QTL, defined as epigenetic QTL, govern the alterations of growth trajectory in a population. Three epigenetic QTL were detected to contribute significantly to variation in growth trajectory during the period from the establishment year to the subsequent year in the field. It is suggested that the activation and expression of epigenetic QTL are influenced by the developmental status of trees and the environment in which they are grown. PMID- 11908666 TI - Microsatellite isolation and characterization in sunflower (Helianthus annuus L.). AB - Development of microsatellite markers for sunflower (Helianthus annuus L.) was performed to estimate their frequency, nature (structure), levels of polymorphism, usefulness for genotype identification, and calculation of genetic relationships between inbred lines representing the species diversity. Isolation was performed from a small-insert genomic library followed by hybridization screening using oligonucleotide probes containing different nucleotide arrays. In this work, 503 unique microsatellite clones were sequenced and 271 PCR primer sequences bordering the microsatellite repeat were designed. For polymorphism assessment, 16 H. annuus germplasm accessions were checked and 170 of the primers tested were shown to be polymorphic for the selected lines. The polymorphic microsatellites produced an average of 3.5 alleles/locus and an average polymorphism information content (PIC) of 0.55. The most frequently found motifs within polymorphic simple-sequence repeats (SSRs) were: (GA)n, (GT)n, (AT)n, followed by trinucleotides (ATT)n, (TGG)n, and (ATC)n, and the tetranucleotide (CATA)n. Most of the 170 SSRs obtained showed important differences in the 16 reference inbred lines used for their characterization. In this work, 20 of the most informative SSRs destined to sunflower genotyping and legal fingerprinting purposes are fully described. PMID- 11908667 TI - Pstl repeat: a family of short interspersed nucleotide element (SINE)-like sequences in the genomes of cattle, goat, and buffalo. AB - The PstI family of elements are short, highly repetitive DNA sequences interspersed throughout the genome of the Bovidae. We have cloned and sequenced some members of the PstI family from cattle, goat, and buffalo. These elements are approximately 500 bp, have a copy number of 2 x 10(5) - 4 x 10(5), and comprise about 4% of the haploid genome. Studies of nucleotide sequence homology indicate that the buffalo and goat PstI repeats (type II) are similar types of short interspersed nucleotide element (SINE) sequences, but the cattle PstI repeat (type I) is considerably more divergent. Additionally, the goat PstI sequence showed significant sequence homology with bovine serine tRNA, and is therefore likely derived from serine tRNA. Interestingly, Southern hybridization suggests that both types of SINEs (I and II) are present in all the species of Bovidae. Dendrogram analysis indicates that cattle PstI SINE is similar to bovine Alu-like SINEs. Goat and buffalo SINEs formed a separate cluster, suggesting that these two types of SINEs evolved separately in the genome of the Bovidae. PMID- 11908668 TI - Identification and characterization of RAPD markers inferring genetic relationships among Pine species. AB - Total genomic DNAs were extracted from several populations of pine species and amplified using oligonucleotides of random sequences. Polymorphism in random amplified polymorphic DNA (RAPD) markers was high and sufficient in distinguishing each of the species. Genetic relationships among eight pine species (Pinus sylvestris, Pinus strobus, Pinus rigida, Pinus resinosa, Pinus nigra, Pinus contorta, Pinus monticola, and Pinus banksiana) from different provenances were analyzed. The degree of band sharing was used to evaluate genetic distance between species and to construct a phylogenetic tree. In general, the dendrogram corroborated the description of relationships based on morphological characteristics and crossability, but also provided new insights into pine taxonomy. RAPD markers specific to some pine species were cloned and sequenced. PCR amplifications using pairs of designed specific primers revealed that all the cloned sequences were likely genus specific because they were not found in spruce or larch. True species-specific sequences were identified using designed primers flanking cloned RAPD fragments. The analysis of RAPD fragment sequences confirmed the genetic relationships among species. A 2281-bp RAPD band called PI-Mt-Stb-23 from P. strobus was used as a probe in restriction fragment length polymorphism (RFLP) analysis and produced distinct banding patterns for each species examined, consistent with the highly polymorphic character of DNA fingerprinting probes. PMID- 11908669 TI - Mosaic haploid-diploid embryos and polyspermy in the tellinid bivalve Macoma balthica. AB - We investigated meiosis, fertilization, and early development in eggs of the tellinid bivalve Macoma balthica (L.), which has external fertilization. Meiosis is standard but polyspermy is found to be very common. In all eight crosses examined, mosaic embryos consisting of a mixture of diploid (2n = 38) and haploid cells occur at a frequency ranging from 2.7 to 29.1%. The earliest mosaic found is in the two-cell stage. We propose that an androgenic haploid cell lineage can originate from one supernumerary sperm that decondenses into a functional haploid nucleus, starts mitosis, and is incorporated in the developing embryo. PMID- 11908670 TI - S1 satellite DNA as a taxonomic marker in brown frogs: molecular evidence that Rana graeca graeca and Rana graeca italica are different species. AB - The brown frog Rana graeca was believed to be present in two areas, the Balkan Peninsula and the Italian Apennines. We have characterised the S1 satellite DNA family from Rana graeca graeca and compared it with that of Rana graeca italica. On Southern blots, the patterns of S1 satellite DNA bands are very different between Italian and Greek specimens, but homogeneous among various populations of the same taxon. The satellite DNA from the Greek taxon contains two repetitive units (S1a (494 bp) and S1b (363 bp)) that could be sequenced after amplification from genomic DNA to directly yield their consensus sequences in each genome. These consensus sequences were very similar among the Greek populations, but differed either in sequence (in S1a) or in both size and sequence (in S1b) from the corresponding repeats of the Italian taxon. A mechanism of concerted evolution is likely responsible for the high homogeneity of S1a and S1b repeat sequences within each genome and species. The genomic content of S1 satellite DNA was lower in the Greek than in the Italian populations (0.5 vs. 1.9%) and fluorescence in situ hybridization (FISH) analysis showed the S1 satellite on only 4 chromosome pairs in the Greek taxon and on all 13 chromosome pairs in the Italian taxon. The completely different structure and genomic organization of the S1 satellite DNA indicate that the Greek and Italian taxa are distinct species: R. graeca and R. italica. PMID- 11908671 TI - DNA polymorphism in the living fossil Ginkgo biloba from the eastern United States. AB - Random amplified polymorphic DNA (RAPD) analysis is a valuable tool in studying inter- and intra-specific genetic variations, patterns of gene expression, and for the identification of specific genes using nearly isogenic variants. Here we used RAPD analysis to study the genetic variation in Ginkgo biloba grown in the eastern United States. Our results support the evidence that Southern blot hybridization of RAPD using probes made from cloned DNA fragments allows a more accurate analysis of the RAPD pattern than dye-stained gels or Southern blot hybridization of RAPD blots using probes made from purified PCR products. Using these techniques, we observed a high degree of relatedness among plants grown in certain localities although significant genetic variation may exist in the species, and could be a possible explanation for the observed variations in the efficacy of medications derived from G. biloba extract. PMID- 11908672 TI - Phylogenetic analysis reveals stowaway-like elements may represent a fourth family of the IS630-Tc1-mariner superfamily. AB - The genomes of plants, like virtually all other eukaryotic organisms, harbor a diverse array of mobile elements, or transposons. In terms of numbers, the predominant type of transposons in many plants is the miniature inverted-repeat transposable element (MITE). There are three archetypal MITEs, known as Tourist, Stowaway, and Emigrant, each of which can be defined by a specific terminal inverted-repeat (TIR) sequence signature. Although their presence was known for over a decade, only recently have open reading frames (ORFs) been identified that correspond to putative transposases for each of the archetypes. We have identified two Stowaway elements that encode a putative transposase and are similar to members of the previously characterized IS630-Tc1-mariner superfamily. In this report, we provide a high-resolution phylogenetic analysis of the evolutionary relationship between Stowaway, Emigrant, and members of the IS630 Tc1-mariner superfamily. We show that although Emigrant is closely related to the pogo-like family of elements, Stowaway may represent a novel family. Integration of our results with previously published data leads to the conclusion that the three main types of MITEs have different evolutionary histories despite similarity in structure. PMID- 11908673 TI - Undermethylated DNA as a source of microsatellites from a conifer genome. AB - Developing microsatellites from the large, highly duplicated conifer genome requires special tools. To improve the efficiency of developing Pinus taeda L. microsatellites, undermethylated (UM) DNA fragments were used to construct a microsatellite-enriched copy library. A methylation-sensitive restriction enzyme, McrBC, was used to enrich for UM DNA before library construction. Digested DNA fragments larger than 9 kb were then excised and digested with RsaI and used to construct nine dinucleotide and trinucleotide libraries. A total of 1016 microsatellite-positive clones were detected among 11 904 clones and 620 of these were unique. Of 245 primer sets that produced a PCR product, 113 could be developed as UM microsatellite markers and 70 were polymorphic. Inheritance and marker informativeness were tested for a random sample of 36 polymorphic markers using a three-generation outbred pedigree. Thirty-one microsatellites (86%) had single-locus inheritance despite the highly duplicated nature of the P. taeda genome. Nineteen UM microsatellites had highly informative intercross mating type configurations. Allele number and frequency were estimated for eleven UM microsatellites using a population survey. Allele numbers for these UM microsatellites ranged from 3 to 12 with an average of 5.7 alleles/locus. Frequencies for the 63 alleles were mostly in the low-common range; only 14 of the 63 were in the rare allele (q < 0.05) class. Enriching for UM DNA was an efficient method for developing polymorphic microsatellites from a large plant genome. PMID- 11908674 TI - Matrix metalloproteinases and cancer. AB - The extracellular matrix (ECM) is a framework of proteins and proteoglycans, secreted by and surrounding stromal fibroblasts. The ECM gives structural integrity to tissues. Remodelling of the ECM is essential for both tumour invasion and angiogenesis. There is abundant evidence to suggest that ECM degrading enzymes, including matrix metalloproteinases (MMPs), are involved in these processes. MMPs are novel targets for therapeutic intervention with the potential to inhibit tumour growth and invasion either on their own or in conjunction with cytotoxic treatments. This review outlines our current understanding of the regulation of MMP expression, the association of MMPs with tumour growth, invasion and angiogenesis and the clinical evolution of MMP inhibitors. PMID- 11908675 TI - Vascular endothelial growth factor and vascular targeting of solid tumors. AB - Vascular targeting agents, which selectively destroy tumor blood vessels, are attractive agents for the treatment of solid tumors. They differ from anti angiogenic agents in that they target the mature, blood-conducting vessels of the tumors. They are better suited for larger tumors where angiogenesis can occur less frequently. For application in man, target molecules are needed that are selectively expressed on the vascular endothelium of tumors. Such markers include the complexes that are formed when vascular endothelial growth factor (VEGF) binds to its receptors (VEGFR). VEGF production by tumor cells is induced by oncogenic gene mutations and by the hypoxic conditions within the tumor mass. The receptors, VEGFR1 (FLT-1) and VEGFR2 (KDR/Flk-1), are upregulated on vascular endothelial cells in tumors by hypoxia and by the increased local concentration of VEGF. Consequently, there is a high concentration of occupied receptors on tumor vascular endothelium. Here, we review the concept of vascular targeting and the development of monoclonal antibodies that bind to VEGF: VEGFR complexes and their use as tumor vascular targeting agents. A promising monoclonal antibody is 2C3, which blocks VEGF from binding to VEGFR2 but not VEGFR1. We conclude that 2C3 might have dual activity as an anti-angiogenic agent by inhibiting VEGFR2 activity and as a vascular targeting agent for selective drug delivery to tumor vessels. PMID- 11908676 TI - Steroids and cytokines in endometrial angiogenesis. AB - Angiogenesis, the formation of new blood vessels from pre-existing ones, occurs physiologically in the endometrium and pathologically e.g. during tumour growth. Sex-steroid hormones affect angiogenesis in the endometrium during the menstrual cycle and might be implicated in cancer angiogenesis. Little information is presently available regarding the exact mechanisms by which these steroids exert their function on the process of both physiological and pathological angiogenesis. In this overview a survey is given of factors important for angiogenesis and of the effects of steroids on the expression of these factors. We have focused on endometrial angiogenesis, because the endometrium is unique in its cyclic growth pattern for which angiogenesis is indispensable. Stimulators and inhibitors of endometrial angiogenesis, that have been found (or suggested) to respond to ovarian steroids, are discussed These factors include vascular endothelial growth factor (VEGF), fibroblast growth factors (FGFs), tumour necrosis factor a (TNF-alpha), erythropoietin (Epo) and trombospondin-1 (TSP-1). Also, the influence of steroids on the expression of matrixdegrading proteases, in particular the plasminogen activator/plasmin system and matrixdegrading metalloproteinases (MMPs) are reviewed, because these proteases play an important role in the migration and invasion of endothelial cells during the process of angiogenesis. An insight into the effects of steroids on endometrial angiogenesis may be helpful to understand and anticipate the potential stimulatory and inhibitory effect of various steroids on angiogenesis in other tissues, in particular tumours. PMID- 11908677 TI - Hepatocyte growth factor expression in human cancer and therapy with specific inhibitors. AB - Hepatocyte growth factor/Scatter Factor (HGF/SF) mediated stimulation of the Met receptor tyrosine kinase results in pleiotropic cellular effects including proliferation, morphogenesis, motility and invasion. In vivo, HGF/SF-Met activation has been shown to participate in tumorigenesis, angiogenesis and metastasis. Coupled with accumulating evidence that aberrant HGF/SF-Met expression is frequently observed in a variety of human tumors, often in association with progressive disease, these data present HGF/SF-Met as an attractive target for therapeutic intervention in human cancer. In this review, we will present the most compelling evidence suggesting a key role for HGF/SF-Met signaling in tumorigenesis, and discuss several possible therapeutic strategies. PMID- 11908678 TI - Fourteen-membered ring macrolides as anti-angiogenic compounds. AB - Macrolide antibiotics have been widely used for infectious diseases since the 1950s. For the last decade, 14-membered ring macrolides, of which erythromycin is the prototype, have attracted a great deal of attention because of their additional therapeutical activities far beyond antibiotics. First, erythromycin has prokinetic effects on the gastrointestinal tract as a motilin receptor agonist. Second, 14-membered ring macrolides, including erythromycin, roxithromycin and clarithromycin have immunomodulating or anti-inflammatory effects in a variety of settings, whereas 16-membered ring macrolides do not. Recently, we found roxithromycin and clarithromycin suppressed angiogenesis and tumor growth in vivo. Both these drugs are administered per os with insignificant side-effects. Their safety has been established through the experience of long term treatment for chronic lower respiratory infectious diseases such as diffuse panbronchiolitis. Although the precise mechanisms have not yet been clarified, 14 membered ring macrolides and their derivatives are promising in therapeutic applications for solid tumors. PMID- 11908679 TI - Assessment of the VEGF, bFGF, aFGF and IL8 angiogenic activity in urinary bladder carcinoma, using the mice cutaneous angiogenesis test. AB - Development of new blood vessels in solid tumors depends on changes in equilibrium between angiogenic stimulators and inhibitors. Overexpression of angiogenic growth factors has been shown in bladder carcinoma. The 'mice cutaneous angiogenesis test' is a good method for assessment of the total angiogenic potential of bladder cancer tissue. The analysis of the levels of proangiogenic factors could be useful for the choice of properly directed angiogenesis inhibitors. The aim of our study was to investigate the influence of blocking some angiogenic factors on the angiogenic activity of bladder cancer tissue. Tumor tissue obtained from 12 patients with invasive bladder carcinoma was used. Cancer tissue homogenates were incubated in the presence of specific antibodies against VEGF, bFGF, Il-8 and aFGF. Cytokine levels were determined using the ELISA test. Cutaneous angiogenesis assay in Balb/c mice was performed to detect the angiogenic activity of the tumor tissue. VEGF, bFGF and Il-8 were present in all examined cancer tissues (aFGF level was not estimated). Cytokine concentration and angiogenic activity of bladder cancer tissue showed individual variation. There was no correlation between the cytokines content in tumor tissue and the ability of this tissue to induce angiogenesis. Absorption caused significant reduction in cytokines level. The reduction of angiogenic activity was observed in the cancer tissue of 1 patient after VEGF absorption, in 3 patients' tissue homogenates after incubation with anti-aFGF and in 2 patients' homogenates after bFGF absorption. There was no reduction of angiogenic activity after Il-8 absorption. PMID- 11908680 TI - Role of angiogenesis in drug resistance. AB - Tumor angiogenesis seems to be an important factor in determining therapeutic effectiveness and patient prognosis. An abundance of data on tumor angiogenesis is available and it clearly shows that most human solid tumors are heterogeneously vascularized and oxygenated and contain hypoxic regions. Such regions and areas of reduced vascularization can affect the response to a variety of drugs. Direct measurements of angiogenesis in various types of tumors have, upon correlation of the data with therapeutic outcome, shown that a high vascular density is associated with a decreased response to therapy and short overall survival time. Therefore, the extent of tumor angiogenesis may well be an important factor contributing to the difficulty of successful therapy in certain tumor types. PMID- 11908681 TI - Vascular effects of photodynamic therapy. AB - Photodynamic therapy (PDT) of tumours is based on a dual selectivity, i.e. preferential uptake and retention of a photosensitiser by tumours and irradiation of the tumour area with light wavelengths specifically absorbed by the photosensitiser. The photoexcited sensitiser generates highly reactive oxygen species that induce irreversible damage to neoplastic cells and vessels. Following sensitiser accumulation and irradiation, damage to sensitive sites within the microvasculature, namely to endothelial cells and the vascular basement membrane, is induced and leads to the establishment of thrombogenic sites within the vessel lumen. This initiates a physiological cascade of responses including platelet aggregation, the release of vasoactive molecules, leukocyte adhesion, increases in vascular permeability and vessel constriction. These result in tumour destruction by vascular collapse, blood flow stasis and tissue hemorrhages. Several photosensitisers are able to induce severe vasculature damage, although by variously different mechanisms. Due to its efficient vascular interactions, photodynamic treatment is also increasingly used for non-cancerous lesions. Successful application of PDT mainly for vessel occlusion and thrombosis in intimal hyperplasia, restenosis, atherosclerotic plaques, corneal and choroidal neovascularisation and port-wine stains is reported. PMID- 11908682 TI - New molecular markers for the study of tumour lymphangiogenesis. AB - The capacity of malignant tumours to metastasize to distant tissues presents a huge problem for the treatment of cancers using conventional surgical and cytotoxic drug therapies. One of the main routes for tumour spread is via the lymphatic vessels, an important conduit for tumours such as breast, lung and gastrointestinal tract that frequently colonize regional lymph nodes. In comparison with the vasculature however, little is known about the biology of tumour lymphatics, tumour lymphangiogenesis or the mechanisms that regulate entry and subsequent migration of tumour cells within lymphatic vessels. This situation has persisted because of the lack of specific molecular markers with which to visualize even normal lymphatics within tissues or to isolate lymphatic endothelial cells for in vitro experimental analysis. Just recently however, novel markers for lymphatic endothelial cells have been identified and their availability has revolutionized research in this field. In this article we highlight the main characteristics of these markers and review recent progress in their use to study tumour lymphangiogenesis. PMID- 11908683 TI - Tumour angiogenesis and response to radiotherapy. AB - The important role of angiogenesis as a predictive factor of response to cytotoxic and radiation therapy has been recently raised. Poor tumour oxygenation is a well recognised feature related to radio-resistance. Since the vascular density is linked to the availability of oxygen and drugs to the tumoural stroma, poor density should be a potent marker of reduced blood perfusion and, therefore, hypoxia and low drug intratumoural concentration. On the other hand, high vascular density and angiogenic ability of cancer is not synonymous with high blood flow since the geometry of the vascular/epithelial component distribution, vascular collapse due to increased interstitial blood pressure, or non-functional vasculature due to an immature structure of the vessels may not allow the establishment of an adequate blood flow, which results in tissue hypoxia. Moreover, activation of angiogenic pathways confer a cancer cell proliferation/apoptosis advantage and trigger an angiogenic regeneration process during fractionated radiotherapy or between the courses of chemotherapy, resulting in rapid tumour re-growth and failure of radiotherapy due to reasons independent of hypoxia and blood flow. The present study reviews the literature on angiogenesis and radiotherapy and suggests a classification of tumours according to their angiogenic ability, which could become a useful tool for the identification of sub-groups of patients that could benefit from specific radiotherapy schedules or combination regimens with cytotoxic and anti-angiogenic compounds. PMID- 11908684 TI - Squamous cell head and neck cancer: evidence of angiogenic regeneration during radiotherapy. AB - BACKGROUND AND PURPOSE: Intra-tumoural neoangiogenesis is an essential process for tumour progression. Although intensification of angiogenic pathways during cytotoxic therapy has been reported by a few experimental studies, the role of angiogenesis in response to radiotherapy is unclear. We recently reported an adverse effect of intense angiogenesis in the radiotherapy outcome of squamous cell head and neck cancer (SCHNC). In the present study we investigated the radiotherapy-induced changes in the microvessel density (MVD) and in the expression of the angiogenic factor thymidine phosphorylase (TP) in SCHNC. PATIENTS AND METHODS: Twenty-four patients with SCHNC underwent a biopsy of the primary lesion immediately before and after delivery of 20Gy of conventionally fractionated radiotherapy. The MVD and the expression of TP was assessed with immunohistochemistry. RESULTS: The irradiated samples were composed of cancer cell islets or bands, immersed within avascular degenerated tissue. In tumours that did not reach complete response after the end of radiotherapy, these viable cancer tissue areas had a significantly higher MVD (p=0.006) and increased percentage of cancer cells with nuclear TP expression (p=0.0004) than the MVD and the TP expression noted in specimens before radiotherapy. TP expression in these islets was directly related to the MVD (p=0.004, r=0.56). CONCLUSION: The present study supports the idea that intensified angiogenic growth (angiogenic regeneration) during radiotherapy is associated with failure of radiotherapy. PMID- 11908686 TI - Tumour hypoxia, hypoxia signaling pathways and hypoxia inducible factor expression in human cancer. AB - Hypoxia has been recognised as an important tumoral feature related to resistance to radiotherapy since 1933. Recent advances in biological research have revealed important aspects on the cellular response to hypoxic stimuli and on the role of hypoxia pathways in the metabolism, growth and progression of cancer. The hypoxia inducible factors (HIF-1a and HIF-2a) have been identified as key proteins that directly respond to hypoxic stress. Following hypoxia, stabilisation and nuclear binding of HIFs triggers the expression of a variety of genes related to erythropoiesis, glycolysis and angiogenesis. This review reports on and discusses the biology of the hypoxia pathways, the studies performed on the expression of HIFs in human cancer and the implications of hypoxia pathways in cancer therapy. PMID- 11908685 TI - Assessment of vascular maturation in lung and breast carcinomas using a novel basement membrane component, LH39. AB - LH39, is a monoclonal antibody recognizing an epitope located at the lamina lucida of mature small veins and capillaries but not in newly- formed vessels of several pathological conditions including cancer. We examined the ratio of mature/immature vessels in 50 breast and 81 lung carcinomas and correlated the vascular maturation index (VMI) to different clinicopathological variables including angiogenesis. Mature vessels were defined by staining with antibodies to both LH39 and CD31, using double immunohistochemistry, whereas immature vessels stained only for CD31. VMI was defined as the percentage fraction of mature vessels (LH39 positive)/total number of vessels (CD31 positive). VMI in breast carcinomas ranged from 0-47% (median 8.75%), which was significantly lower than that observed in the normal breast cases (range 54%-70%; median 68%). The median VMI in the non-small cell lung carcinomas was 46% (range 15%-90%). There was a significant inverse correlation between high tumor VMI and absence of nodal involvement in both breast and lung tumors examined (p=0.01). Thymidine phosphorylase (TP) expression, but not vascular endothelial growth factor (VEGF) expression, was related to a low VMI showing an intense vascular remodeling in TP expressing cases. Thus, assessment of vessel maturation might be complementary to microvessel number to aid the identification of patients who might benefit from specific antiangiogenic therapies or vascular targeting treatment. PMID- 11908687 TI - The prognostic role of angiogenesis in breast cancer. AB - Angiogenesis is of key importance in the process of tumour progression in a number of tumour types including breast cancer. Breast cancer angiogenesis has been the most extensively studied and now serves as a paradigm for understanding the biology of angiogenesis and its effects on tumour outcome and patient prognosis. The first study to examine intra-tumoural microvessel density (IMD) immunohistochemically, and relate it to tumour outcome, was carried out by Weidner and colleagues in 1991 using an antibody against factor-8 related antigen as an endothelial marker in a series of breast cancers. They found a near linear relationship between increased microvessel counts and metastasis. This work defined the standard methodology still used today for the morphometric assessment of IMD, such that by evaluating only three so called "hotspot" areas it was possible to determine the metastatic potential of a tumour. The biological rationale behind this was that these highly angiogenic areas were those most likely to be the easiest point of entry for tumour cells into the systemic circulation. Since this initial work there have been many studies which confirm these findings and have related angiogenesis to poor prognosis using a variety of antibodies including those to CD31 and CD34. Angiogenesis is also potentially a unique target for anti-tumour therapy, and much research is being carried out in this area, including blockade of angiogenic signalling pathways and the therapeutic use of antiangiogenic factors. PMID- 11908688 TI - Angiogenesis, angiogenic factor expression and hematological malignancies. AB - Solid tumor growth consists of an avascular and a subsequent vascular phase. Several studies have now shown that, as in solid tumors, angiogenesis also plays a critical role in the progression of hematological malignancies. Monoclonal gammopathy of undetermined significance (MGUS) and non-active to active multiple myeloma (MM) progress when plasma cells induce angiogenesis and this in turn promotes progression. The increased bone marrow neovascularization, increased angiogenic and proteolytic potential of plasma cells may explain the frequent occurrence of extramedullary localization in MM. As observed in active MM, enhanced bone marrow neovascularization is apparent in acute untreated lymphoblastic leukemia. In B-cell non-Hodgkin's lymphomas, angiogenesis is significantly enhanced in relation to progression. Angiostatic molecules, such as thalidomide, could also be considered for the clinical management of hematological tumors. PMID- 11908689 TI - Prognostic significance of angiogenesis in rectal cancer: a morphometric investigation. AB - BACKGROUND: Neo-angiogenesis is crucial for tumor growth and metastasis and has been proposed as an independent prognostic factor for survival in patients with solid tumors. In this study the quantitative expression of angiogenesis was investigated by direct stereologic assessment of the vascular surface density in rectal carcinoma to determine the possible correlation of angiogenesis with clinicopathological factors and prognosis. PATIENTS AND METHODS: Sections from formalin-fixed paraffin-embedded tissue blocks of 29 primary rectal carcinomas were resected and immunostained for endothelial cell factor-VIII-related antigen. The vascular surface density (VSD), number of vessels per square mm (NVES), maximum NVES (NVESmax) according to the three maximum values of NVES and number of vessels in the unit area (N) were assessed by means of morphometry. The results were related to the main prognostic variables and the survival of patients. RESULTS: There were no significant differences between survivors and non-survivors in terms of the angiogenesis parameters that were investigated. The overall survival rate was not significantly different for sex, age, tumor size and differentiation, extrahepatic metastasis, depth of invasion and the mode of adjuvant therapy. However, a significantly lower overall survival rate was observed in patients with liver metastatic disease (p<0.001), lymph node involvement (p=0.04) and incomplete resection (p<0.001). Multivariate analysis indicated that only the number of vessels in the unit area (HR = 1. 028, p = 0. 04), hepatic metastases (HR=14.94, p=0.007) and type of resection (HR=23.81, p=0.004) predicted overall survival. CONCLUSION: These findings suggest that increased tumoral vascularity, consistent with previous studies, adversely affects survival in rectal cancer patients. Liver metastatic status and completeness of the surgical resection were the most powerful criteria to predict the final outcome of these patients. Thus, neo-angiogenesis is indeed an important and key step in tumorigenesis, but it may not be the single overwhelming factor that determines recurrence and metastasis in rectal carcinoma. PMID- 11908690 TI - Prognostic role of angiogenesis in colorectal cancer. AB - The need for dependable prognostic markers in colorectal cancer, both in the advanced as well the as adjuvant setting, is greater than ever. The introduction of new chemotherapeutic agents in the clinic, with different mechanisms of action as well as different side-effect profiles, has made this even more important. Angiogenesis, the formation of new vessels, is essential for tumour growth and metastasis. The adverse impact of tumour angiogenesis in the context of colorectal cancer on relapse and prognosis has been evaluated in numerous retrospective studies. Most of these studies have tended to be relatively small in terms of patient numbers and sometimes have reached conflicting conclusions. A number of different angiogenic factors, including vascular endothelial growth factor (VEGF), thymidine phosphorylase (TP) and intratumoural microvessel density (IMD), have been assessed to varying extents. This article highlights some of the important developments in this rapidly expanding field. PMID- 11908692 TI - Angiogenesis, angiogenic factor expression and prognosis of gastric carcinoma. AB - It is known that the growth and metastasis of malignant tumors depends on neovascularization. It has also been suggested that the degree of tumor angiogenesis is related to clinical outcome in several tumor types. This is true for gastric carcinoma, where tumor angiogenesis is closely correlated with prognosis and hematogenous metastasis. Several types of angiogenic factors have been investigated in gastric cancer. In the current review, the correlation between angiogenesis / angiogenic factor expression and prognosis in gastric cancer is discussed. Moreover, the potential clinical applications of antivascular, anti-angiogenic and angiostatic agents for the treatment of gastric carcinoma are summarized. PMID- 11908691 TI - Angiogenesis, angiogenic factor expression and prognosis of bladder cancer. AB - Angiogenesis is the process by which tumours induce a blood supply from their surrounding tissues and it has been shown to be necessary for tumour growth. Evidence is accumulating for both the prognostic usefulness of measures of angiogenesis and its potential as a target for anticancer therapy. This review discusses the evidence concerning the association between angiogenesis and bladder cancer, focusing on the mechanisms behind the angiogenic process and the quantification of factors believed to be involved, relating these clinically to their prognostic use and to the antiangiogenic strategies so far described in vitro and in vivo. PMID- 11908693 TI - Prognostic role of angiogenesis in non-small cell lung cancer. AB - The prognostic role of angiogenesis has been thoroughly examined during the past decade. Using specific monoclonal antibodies which recognize the endothelium and counting the microvessel density (MVD) under optical microscope, the tumour angiogenic ability can be estimated. In non-small cell lung cancer (NSCLC) a large number of studies have provided significant evidence that high MVD is one of the most important and independent variables defining poor outcome after surgery. The development of antibodies against several angiogenic factors, working on paraffin-embedded material, allowed the study of the angiogenic and prognostic role of molecules involved in the angiogenic process. Most of the studies published in the literature agree that expression of VEGF (vascular endothelial growth factor) and of thymidine phosphorylase are important prognostic factors in NSCLC. This review analyzes the results of clinicopathological studies on the prognostic role of MVD and of angiogenic factor expression in NSCLC. PMID- 11908694 TI - Angiogenesis and endometrial cancer. AB - Angiogenesis occurs more frequently in endometrial carcinomas developing against a background of endometrial atrophy rather than carcinomas arising from a hyperplastic endometrium. In some studies, angiogenesis is associated with unfavourable histopathological features, but not in others. In all studies, however, increased angiogenesis is related to poor prognosis. Vascular endothelial growth factor (VEGF) is the major stimulus for endothelial cell proliferation in endometrial carcinomas and is, therefore, associated with high angiogenesis. VEGF expression at the invading tumour front is 4-10 times higher than in the inner tumour areas and is significantly associated with poor prognosis, particularly within stage I endometrial disease. However, since its stimulating effect on endothelial cells is basically dependent on the presence of VEGF receptors, i.e. the flk-1(KDR), the detection of a functionally intact angiogenic pathway VEGF/flk-1 (KDR) is a more reliable and, indeed, independent prognostic parameter. The other angiogenic factor, thymidine phosphorylase (TP), is related to the adverse histopathological variables of the non endometrioid carcinomas, high tumour grade, deep myometrial invasion and advanced stage of disease, at least at the invading tumour front, and its prognostic significance is, therefore, limited. Furthermore, it is not directly related with increased angiogenesis. However, the simultaneous expression of high VEGF / high TP activity at the invading tumour front emerged as the most potent angiogenic phenotype in endometrial carcinomas, indicating that the two molecules are co operating. Furthermore, a high TP activity in the stromal cells is associated with a high density of activated macrophages which further promotes endometrial tumour angiogenesis. PMID- 11908695 TI - Biochemical decompression of hydrogen by naturally occurring bacterial flora in pigs: what are the implications for human hydrogen diving? PMID- 11908696 TI - Exposure to hyperoxia in diving and hyperbaric medicine--effects on blood cell counts and serum ferritin. AB - A reduction in hemoglobin concentration has been consistently reported after deep saturation dives, whereas reductions in thrombocyte counts and changes in biochemical parameters specific for liver function have been reported after some dives. In this study the contribution of exposure to hyperoxia to these changes were studied. Hemoglobin concentration, blood cell counts, serum ferritin, and biochemical parameters specific for liver damage were measured before and after a shallow 28-day saturation dive to a pressure of 250 kPa with the same hyperoxic exposure (40-50 kPa) as in a deep saturation dive in eight male divers. The same parameters were measured before, during, and after a standard 21-day hyperbaric oxygen (HBO2) treatment series in a selected group of 16 patients (8 male). There were significant reductions in hemoglobin concentrations of 3.8 +/- 4.7% (P = 0.023) and 10.2 +/- 5.3% (P = 0.003) after the HBO2 treatment series and dive, respectively, accompanied with reductions in red cell counts, reticulocyte counts, and hematocrit. There was an increase in ferritin concentrations of 29 +/ 21% (P = 0.002) and 107 +/- 43% (P < 0.001). In contrast to some deep dives, there were no changes in thrombocyte counts or biochemical parameters specific for liver damage. Exposure to hyperoxia contributes significantly to reduced hemoglobin and increased ferritin concentrations after saturation dives. The changes may reflect a shift of iron from synthesis of hemoglobin in the bone marrow to storage in macrophages caused by a downregulation of hemoglobin synthesis, or an increased oxidative stress. The changes are too small to be of clinical significance with respect to diving and HBO2 treatment. PMID- 11908697 TI - Effects of inspiratory and expiratory resistance in divers' breathing apparatus. AB - This study was performed to determine if inspiratory breathing resistance causes greater or smaller changes than expiratory resistance. Unacceptable inspiratory resistances were also determined. Five subjects exercised at 60% of their VO2max while immersed in a hyperbaric chamber. The chamber was pressurized to either 147 kPa (1.45 atm abs, 4.5 msw, 15 fsw) or 690 kPa (6.8 atm abs, 57 msw, 190 fsw). Breathing resistance was imposed on the inspiratory or expiratory side and was as high as 0.8-1.2 kPa liter(-1) x s(-1) (8-12 cm H2O x liter(-1) x s(-1)) at a flow of 2-3 liter x s(-1) at 1 atm abs., the other side being unloaded. The subjects reacted to the imposed load by prolonging the phase of breathing that was loaded. Inspiratory breathing resistance caused greater changes than expiratory resistance in end-tidal CO2, dyspnea scores, maximum voluntary ventilation, and respiratory duty cycle. Using previously published criteria for acceptable levels of dyspnea scores and the CO2 levels, we found that an inspiratory resistance inducing a volume-averaged pressure of 1.5 kPa is not acceptable. Similarly, an expiratory resistance should not induce a volume-averaged pressure exceeding 2.0 kPa PMID- 11908698 TI - Effects of water immersion on pulmonary function in asthmatics. AB - Immersion induces air trapping in the lungs, as does asthma. Consequently, when using diving apparatus, asthmatics may face greater risk than non-asthmatics of pulmonary barotrauma (PBT) during ascent. We studied the pulmonary airflows and closing capacities (CC = closing volume + residual volume) in subjects with exercise-induced asthma (A, n = 12) and in healthy controls (C, n = 11) under four conditions: dry and immersed, both before and after exercise (treadmill running, non-immersed). Immersed, both C and A had significant and equivalent reductions in vital capacity, FEV1, FEV1/FVC, and FEF25%-75%. Post-exercise and immersed, pulmonary airflows deteriorated further in A but were better in C: FEV1 (A, 3.6 +/- 0.8 liter vs. 3.3 +/- 0.8 liter, P = 0.001; C, 3.9 +/- 0.5 liter vs. 4.1 +/- 0.6 liter, P = 0.006), FEF25-75% (A, 3.5 +/- 1.0 liter x s(-1) vs. 3.0 +/ 0.8 liter x s(-1). P < 0.05; C, 4.0 +/- 0.9 liter x s(-1) vs. 4.3 +/- 0.9 liter x s(-1), P < 0.05). Therefore, in contrast to C, A subjects had reduced pulmonary airflows during immersion after exercise. Furthermore, A subjects more often had no closing volume phase IV when immersed after exercise than C (P = 0.005). Interpreting the absence of phase IV as indicative of more air trapping in the asthmatics during immersion after exercise would be consistent with the reductions in airflow. PMID- 11908699 TI - Dysbaric osteonecrosis in Turkish sponge divers. AB - Skeletal radiographs were performed to determine the prevalence of dysbaric osteonecrosis (DON) in 51 Turkish sponge divers. DON was correlated with the diver's age and experience, maximum diving depth, and decompression sickness (DCS). Thirty-six of the 51 divers had radiographic evidence of one or more lesions, for a prevalence of 70.6% DON. Proximal humerus was the most effected site. Type B (head, neck, and shaft) lesions were most common, comprising 63.6% of all DON lesions. Type A (juxta-articular) lesions were observed in all 10 divers who complained of painful motion of their shoulder or hip joints. DON most commonly affected the proximal humerus. Reportedly, 38 of these 51 (74.5%) divers had experienced DCS. We did not fnd any significant relationship between DON and DCS, maximum diving depth, diving experience, and divers' age. PMID- 11908700 TI - Decompression sickness risk reduced by native intestinal flora in pigs after H2 dives. AB - Decompression sickness (DCS) risk following a simulated dive in H2 was lower in pigs with a native intestinal flora that metabolized H2. Pigs (n = 27; 19.4 +/- 0.2 kg body mass) were placed in a chamber that was pressurized to 22.2-25.5 atm (absolute; 2.2-2.6 MPa) with 84-93% H2 for 3 h. Chamber concentrations of O2, H2, He, N2, and CH4 were monitored by gas chromatography. Release of CH4 from the pigs indicated that intestinal microbes had metabolized H2 After decompressing to 11 atm, the pigs were observed for DCS. Animals with DCS released significantly less (P < 0.05) methane (0.53 +/- 0.37 ppm CH4; n = 5) than those without DCS (1.40 +/- 0.17 ppm CH4; n = 22). The DCS risk reduction was attributed to the loss of roughly 12% of the total volume of H2 that could be stored in the tissues of the pigs. Thus, H2 metabolism by the native intestinal flora of pigs may protect against DCS following a simulated H2 dive. PMID- 11908701 TI - The role of NK cells in autoimmune disease. AB - NK cells are a subset of mononuclear cells which have long been suspected of playing an immunoregulatory role in the prevention of autoimmune diseases. Here, we briefly discuss the characteristics of NK cells--particularly what is known of their functional capabilities--and summarise the major findings from studies of NK cells in human and animals susceptible to three major autoimmune diseases: multiple sclerosis, systemic lupus erythematosus and type 1 (autoimmune) diabetes mellitus. In each case, we present the evidence for an association between disease and deficiencies in NK cells. The prospect of clinical interventions that stimulate NK cell activity are discussed and the current status described. PMID- 11908702 TI - Soluble Fas ligand-susceptible "memory" cells in mice but not in human: potential role of soluble Fas ligand in deletion of auto-reactive cells. AB - Human soluble Fas ligand (sFasL) has an apoptotic activity in contrast to murine sFasL. The physiological function of human sFasL is not known, while the pathological consequence of sFasL overproduction has been reported. To understand the physiological function of (human) sFasL, murine and human lymphocytes were treated with sFasL. sFasL treatment significantly decreased CD45RBlo "memory" CD4+ lymphocyte fraction and increased propidium iodide (PI)+ apoptotic CD45RBloCD4+ lymphocytes among murine peripheral lymphocytes. However, sFasL treatment neither decreased CD45RO+ "memory" CD4+ lymphocyte fraction nor increased PI+ CD45RO+CD4+ lymphocytes among human peripheral lymphocytes, suggesting that the deletion of memory cells by sFasL had already occurred in vivo. Patients with systemic lupus erythematosus had sFasL-susceptible "memory" cell fraction suggesting an incomplete deletion of such "memory" cells. These results suggest that the physiological function of human sFasL is to delete the potentially auto-reactive "memory" lymphocytes, which complements membrane FasL (mFasL)-mediated deletion of auto-reactive cells in human beings but not in mice. PMID- 11908704 TI - Evidences of the involvement of Bak, a member of the Bcl-2 family of proteins, in active coeliac disease. AB - Apoptosis of enterocytes is a feature that characterises the development of lesions in coeliac disease (CD). However, the intracellular pathways that lead to apoptosis of enterocytes have not been completely clarified. Bak is a member of the Bcl-2 family of proteins that acts as an endogenous promoter of apoptosis in normal enterocytes. However, its role in coeliac lesions has not been explored. We used small intestinal mucosa from patients with CD to evaluate the differential expression of members of the Bcl-2 family of proteins. Gene expression of Bak was analysed by RT-PCR of biopsies from 14 patients with untreated CD and from 19 controls without CD. In these samples, we also investigated the localisation of the Bak protein by immunohistochemistry and its apoptotic activity. In patients with untreated CD there was a 2.3-fold higher expression of Bak mRNA (p = 0.026), without significant differences in the expression of related genes bax or bcl-2. The higher expression of interferon gamma (IFN-gamma) (p = 0.036) and the higher number of apoptotic cells identified by the TUNEL method (p = 0.032) confirmed the proapoptotic status in the intestinal mucosa of CD patients. We found a significant positive correlation (p < 0.0001) between the expression of IFN-gamma and Bak mRNA in patients with untreated CD. The expression of Bak protein was higher in patients with CD, and the immunoreactivity was almost restricted to the epithelium. We found that Bak mRNA and its protein were overexpressed in the intestinal lesions of CD patients and that IFNgamma confers increased susceptibility for enterocytes to undergo apoptosis via upregulation of Bak. PMID- 11908703 TI - Inhibitory effects of incomplete Freund's adjuvant on experimental autoimmune encephalomyelitis. AB - Freund's incomplete adjuvant (IFA), an aqueous/oil emulsion that is widely used in combination with antigenic proteins and peptides to induce tolerance, is considered to be immunologically inert. However, sporadic reports indicate that IFA may itself have inhibitory properties on induction of adjuvant induced arthritis and spontaneous diabetes. In the current study, the effects of IFA/saline were evaluated on the induction of experimental autoimmune encephalomyelitis (EAE) in three different strains of mice. IFA/saline given i.p. in two doses of > 100 microl 10 days apart were found to inhibit EAE induction to varying degrees in all three strains of mice in a dose dependent fashion. The IFA/saline injections inhibited both mitogen and antigen-induced T cell proliferation, induced elevated secretion of IFN-gamma and IL-10 by neuroantigen specific T cells, and reduced expression of cytokines, chemokines, and chemokine receptors of CNS-infiltrating mononuclear cells. These data demonstrate for the first time a direct inhibitory effect of IFA/saline on EAE, and re-emphasize the need to properly control experiments using IFA to induce antigen-specific tolerance. PMID- 11908705 TI - Th1 and Th2 cytokine modulation by IL-10/IL-12 imbalance in autoimmune haemolytic anaemia (AIHA). AB - Recent studies about autoimmune diseases in animal models and in humans focused their attention on lymphocyte activation and in vitro cytokine production. The respective contribution of the Th1 and Th2 cytokines to the pathogenesis of autoimmune diseases is still a matter of debate. In this study the role of IL-2, IL-4, IFN-gamma, IL-10 and IL-12 cytokines were investigated by examining their spontaneous and mitogen-induced (OKT3 and PHA or LPS) synthesis and T-cells proliferative response by peripheral blood mononuclear cells to determine their role in the pathogenesis of AIHA. Thirteen patients affected by AIHA, idiopathic or associated with other diseases, and 13 healthy subjects, randomly selected from a group of blood donors, were investigated. This study indicated that AIHA is characterised by increased basal synthesis of IL-4 and decreased levels of IFN gamma compared with healthy controls (p < 0,01). These results suggest that there is a basal decrease of Th1 cytokine and an increase of the Th2 ones. Enhanced IL 2 levels in AIHA patients are likely due to the necessity of a T-cell proliferation stimulus rather than produced as Th1 prevalent stimulation. Furthermore, it has been observed a significant increase in IL-12 production in LPS stimulated cultures from healthy controls, but not in AIHA patients, that shows IL-10 increased levels, which could cause a secondary decrease in IFN-gamma production and a stimulation of Th2 differentiation. These observations indicate that decreased production of Th1-type cytokines and prevalent Th2 ones leading to autoantibodies production in AIHA may be secondary to the imbalance between IL-10 and IL-12. These results strongly suggest that manipulation of the cytokine network, i.e. IL-10/IL-12 balance, maintained by cells of the innate immune system, can have a strong effect on the incidence of AIHA and their modulation might be useful for a therapeutic control of the disorder. PMID- 11908706 TI - Specific proliferation towards myelin antigens in patients with multiple sclerosis during a relapse. AB - Whether autoreactive T cells from multiple sclerosis (MS) patients display a certain autoreactive pattern is controversial. In this study, we have analyzed reactivity towards myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), alpha B-crystallin and S100beta antigens in 35 relapsing remitting MS patients and 12 healthy controls (HC). During relapse, we observed T specific proliferation towards MBP (15.8%), MOG (38.9%), alpha B-crystallin (11.1%) and S100beta (26.3%) in MS patients. Reactivity to MBP (12%), MOG (28%), alpha B-crystallin (28%) and S100beta (19.2%) was also observed in HC. There were changes in the specific proliferation in consecutive samples obtained from either patients or HC. Such fluctuations did not follow any specific or conservative patterns. Antigen-specific cytokine production was also assessed as a method to evaluate whether there were differences in the qualitative response between MS patients and HC, with negative results. In summary, we show here that the reactivity patterns, as measured by specific proliferation and cytokine production, are similar in RR-MS patients and HC and fluctuate over time. PMID- 11908707 TI - Plasmapheresis modulates Th1/Th2 imbalance in patients with systemic lupus erythematosus according to measurement of intracytoplasmic cytokines. AB - To examine the possible effect of plasmapheresis on the ratio of Th1/Th2 type cytokine-secreting cells we recruited eight patients with active systemic lupus erythematosus into the present study. They all failed to respond to conventional therapy. A sensitive multiparametric flow cytometric analysis was used for the detection of intracellular IL-4, IL-10 and IFNgamma. Stimulated peripheral blood cells were analysed by this procedure. Plasmapheresis was performed every second day for three occasions, using a continuous flow type blood cell separator, and a total of 100 ml/body weight kg plasma was removed. Patients received 1 mg/kg/day methylprednisolone during this period. As a result of the procedure, the rate of IFNgamma positive Th cells increased, while the rate of IL-4 and IL-10 expressing CD4 positive cells decreased. Together with these observations the concentration of anti-ds-DNA antibodies decreased after plasmapheresis. A decrease in disease activity index (SLE-DAI) indicated the clinical effectiveness of the therapy. PMID- 11908708 TI - Effect of diesel exhaust particle extracts on collagen-induced arthritis in mice. AB - We investigated the effect of diesel exhaust particles (DEP) extracts on collagen induced arthritis (CIA) in mice. For this study, a single DEP sample was consecutively extracted with hexane (HEX-DEP), benzene (BEN-DEP), dichloromethane (DIC-DEP), methanol (MET-DEP), and 1 M ammonia (AMM-DEP) in that order. Residues unextracted with the last extraction solvent 1 M ammonia (UNE-DEP) were also used for experiments. To induce CIA, mice were immunized with type II collagen (CII) and 21 days later given a booster injection. DEP, each DEP extract, and UNE-DEP were intranasally administered every two days over a period of 20 days, commencing on the day of immunization. The results showed that treatment of mice with DEP, DIC-DEP, and UNE-DEP augmented both the incidence and the severity of CIA. DEP and DIC-DEP increased production of anti-CII IgG, IgG2a, and IgG1 antibodies as well as secretion of JFN-gamma and IL-4. Treatment with UNE-DEP resulted in an increase in antigen-specific IgG, IgG2a, and IFN-gamma but neither IgG1 nor IL-4. AMM-DEP failed to affect CIA as well as production of IgG2a and IFN-gamma, although significant increases in anti-CII IgGI and IL-4 were observed in the treatment group. HEX-DEP, BEN-DEP, and MET-DEP had no effects on CIA and production of antibodies and cytokines examined. Thus, DEP and DIC-DEP appear to contain compounds, which enhance both Th1 and Th2 responses, while UNE-DEP and AMM-DEP to contain chemicals, which augment Th1 and Th2 alone, respectively. Th1- but not Th2-modulating compounds from DEP, DIC-DEP, and UNE-DEP seem to influence CIA. PMID- 11908709 TI - Sequence analysis of Tnf as a candidate for Idd16. AB - Linkage analysis and congenic mapping have localized 18 loci (Idd1-18) that contribute to the development of autoimmune type 1 diabetes in the nonobese diabetic (NOD) mouse. By using a congenic NOD strain which possesses recombinant MHC from a closely related CTS strain, a susceptible region (Idd16) was mapped to the segment adjacent to, but distinct from class II A and E genes (Idd1). The tumor necrosis factor alpha gene (Tnf), which is located within the Idd16 region, has been suspected to be a candidate gene for type 1 diabetes in the NOD mouse. Although the protein-coding region in Tnf has been sequenced in the NOD mouse and its related strains, the complete upstream region (approximately 1400 bp, including the 5'-untranslated region) has not yet been studied. To study the possible contribution of the transcriptional regulation of Tnf to susceptibility to type 1 diabetes, we determined the complete nucleotide sequences of the NOD strain and its related strain, CTS, in comparison with the non-diabetic control strain, C57BL/6. The nucleotide sequence of the 5'-upstream region in the NOD mouse was identical to that in the C57BL/6 mouse, but different from that in the CTS mouse. In particular, a C to A substitution at position 3408 in the CTS mouse creates a new GATA family binding site, which may be responsible for the lower incidence of type 1 diabetes in the NOD. CTS-H-2 congenic strain despite the presence of the same class II MHC. PMID- 11908710 TI - Organ-specific autoimmunity in granulocyte macrophage-colony stimulating factor (GM-CSF) deficient mice. AB - Granulocyte macrophage-colony stimulating factor (GM-CSF) is a pleiotrophic proinflammatory cytokine that augments adaptive immunity by linking it to innate immunity. Experimental autoimmune gastritis is an animal model of human autoimmune gastritis and pernicious anaemia. We have previously shown that GM-CSF is expressed in the gastric mucosa of mice with gastritis initiated by neonatal thymectomy (Gastroenterology 110 (1996) 1791) and that transgenic expression of GM-CSF in the stomach induces autoimmune gastritis in mice (J. Immunol. 166 (2001) 2090). Here we have examined the development of autoimmune gastritis initiated by immunisation or by neonatal thymectomy in GM-CSF deficient mice. We found that gastritis develops in GM-CSF deficient mice initiated by neonatal thymectomy but not by immunisation with gastric antigen. These observations suggest that GM-CSF is not absolutely required for the initiation of autoimmunity and highlights the different conclusions that can be drawn using different disease models. PMID- 11908711 TI - Current management of polycythemia vera. AB - Over a century has elapsed since the first description of polycythemia vera (PV), and current treatment recommendations are primarily based on the results of clinical trials that were performed in the late 1960s and early 1970s. Continued identification and appropriate utilization of PV-specific biologic parameters may allow substantial modification of early diagnostic criteria. New cytoreductive treatment agents are increasingly being used without any evidence of superiority over conventional therapy. The role of aspirin is being readdressed by an ongoing controlled study. Transformation of PV into either myelofibrosis with myeloid metaplasia or acute leukemia remains a major complication that may not be influenced by current therapy. PMID- 11908712 TI - Results of a randomized phase III trial in children and adolescents with advanced stage diffuse large cell non-Hodgkin's lymphoma: a Pediatric Oncology Group study. AB - PURPOSE: The Pediatric Oncology Group (POG) adopted a histology-based approach to the management of pediatric non-Hodgkin's lymphomas (NHL) utilizing the National Cancer Institute Working Formulation for Clinical Usage. Patients with diffuse large cell lymphoma (DLCL) were treated on a separate protocol from small cell diffuse undifferentiated or lymphoblastic lymphomas. This study assessed the overall and event free survival of children with DLCL and determined the effects of cyclophosphamide upon these end-points in a prospective randomized trial. PATIENTS AND METHODS: One hundred and twenty eligible stage III or IV NHL patients with the confirmed diagnosis of diffuse large cell or immunoblastic histology were enrolled on study between October 1986 and November 1991. Patients were randomized to receive or not receive cyclophosphamide: 58 received cyclophosphamide, doxorubicin, vincristine, 6-mercaptopurine (6-MP), and prednisone (ACOP+) and 62 were treated with doxorubicin, vincristine, 6-MP, and prednisone (APO). In both treatment programs methotrexate was substituted when the doxorubicin cumulative dose reached 450 mg/m2. Radiation was administered to bulky disease if progression or no response were observed after induction therapy. Planned duration of therapy was 12 months. RESULTS: The 5-year event free survival (EFS) rates of patients treated with ACOP+ versus APO were 62+/-7 and 72+/-6%, respectively. While there was no statistically significant difference between the two treatment arms (p = 0.28), we can only say that we are 95% confident that the difference in 5-year EFS falls in the wide range from 28% in favor of APO to 8% favoring ACOP+. Marrow suppression was the main toxicity with one fatal infection. There were three other deaths on study due to respiratory failure in patients with mediastinal masses. Only one patient experienced cardiotoxicity requiring discontinuation of doxorubicin. Ten patients received radiation therapy to achieve remission. CONCLUSION: The efficacy of elimination of cyclophosphamide from the treatment program of children and adolescents with advanced stage diffuse large cell lymphoma was inconclusive as to its effect on EFS. Furthermore, the majority of the patients (92%) did not require any radiation therapy to bulky disease indicating that the chemotherapy regimens are quite efficient for achievement of complete remission. PMID- 11908713 TI - Primary treatment of low-grade non-Hodgkin's lymphoma with the combination of fludarabine and mitoxantrone: a phase II study of the Hellenic Cooperative Oncology Group. AB - The treatment of patients with recurrent low-grade lymphoma with the combination of fludarabine, mitoxantrone and dexamethasone has been associated with significant activity but has also caused frequent infectious complications. We designed a phase II study for previously untreated patients with the combination of fludarabine and mitoxantrone but without steroids. Our aim was to assess the activity of this combination as primary treatment for low-grade lymphoma and to avoid the additional immunosuppression induced by dexamethasone. Twenty seven patients with low-grade lymphoma received fludarabine 25 mg/m2/day i.v. on days 1 3 and mitoxantrone 10 mg/m2 i.v. on day 1. The treatment was repeated every 28 days for a maximum of six cycles. Twenty patients (74%) achieved an objective response including 12 (44%) complete and 8 (30%) partial responses. The main toxicity was grade III or IV neutropenia, which occurred in 40% of patients but there were no severe opportunistic infections. The median time to progression for all patients was 32 months. With a median follow-up of 33.4 months, six patients have died and the probability of survival at 3 years is 75%. We conclude that the fludarabine and mitoxantrone regimen is safe and effective for newly diagnosed patients with low-grade lymphoma who require treatment. Prospective randomized trials are needed in order to assess the impact of this treatment on patients' survival. PMID- 11908714 TI - Breast lymphoma associated with breast implants: two case-reports and a review of the literature. AB - Breast lymphoma is a rare entity consisting mostly of B-cell lymphomas that affects older women. Very few cases of breast lymphomas of T-cell origin have been reported. Even fewer cases of breast lymphomas have been reported in women who have received breast implants. Silicone, a frequent component of breast implants, has been extensively investigated as a possible etiologic agent for some neoplasias and autoimmune disorders, including non-Hodgkin's lymphoma. We herein report two unusual cases of anaplastic large cell lymphoma of T-cell phenotype developing in the breasts of women who had received breast implants. PMID- 11908715 TI - Treatment of refractory peripheral T-cell lymphoma with denileukin diftitox (ONTAK). AB - Peripheral T-cell lymphomas (PTCLs) are an uncommon and heterogeneous group of well-differentiated, post-thymic T-cell malignancies that can present in the skin as cutaneous T-cell lymphomas. In general, their prognosis is poor, and specific therapy is not well defined. We report the successful treatment of a patient with relapsed, refractory PTCL who after failing 13 standard single and multiple chemotherapy regimens and experimental agents had a dramatic prolonged response to diftitoxin denileukin (ONTAK). This fusion protein, composed of diphtheria toxin coupled to interleukin-2, is approved for cutaneous T-cell lymphomas, including mycosis fungoides, and should be considered for treatment of the rare subset of peripheral T-cell lymphomas. PMID- 11908717 TI - Angioimmunoblastic lymphadenopathy (AILD) may respond to thalidomide treatment: two case reports. AB - The histological morphology of AILD-type T-cell lymphoma shows proliferation of small, arborizing high endothelial venules. Patients typically have generalized lymphadenopathy, fever, weight loss, skin rash, polyclonal hypergammaglobulinemia and autoimmune phenomena, and are susceptible to infections. The clinical course is usually aggressive. About 30% of patients have an initial remission on steroids alone, but most require some form of cytotoxic chemotherapy. Median survival ranges from 15 to 24 months, and it appears that the disease cannot be cured. We have used anti-angiogenetic therapy with thalidomide in two AILD patients and observed remarkable responses. PMID- 11908716 TI - Adult T-cell leukemia/lymphoma: a rare case in the USA and review of the literature. AB - Adult T-cell leukemia/lymphoma (ATLL), in its acute stage, is a uniformly fatal disease. ATLL is caused by the human T-cell lymphotropic virus I (HTLV-1), a retrovirus endemic in numerous areas throughout the world including Japan, the Caribbean, Central and South America and certain areas of the United States. Although the progression from HTLV-1 carrier status to ATLL occurs only rarely, ATLL is incurable and thus prevention of HTLV-1 transmission should be a primary goal. With the development of new anti-retroviral and monoclonal therapies, there exist potential cures or at least prolonged remissions for patients diagnosed with ATLL. We present a case of ATLL that, to our knowledge, is only the third reported case in Georgia. In addition, we present a brief review of the literature, including potential new treatment regimens that appear to have promise in the treatment of ATLL. PMID- 11908718 TI - Vascular endothelial growth factor (VEGF) expression in plasmacytoma. AB - Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis. Although the role and importance of angiogenic factors such as VEGF have been established in various solid tumors, this has not been widely evaluated in hemopoietic neoplasias. In this trial, VEGF was studied in plasmacytoma and VEGF expression was compared with histopathologic grade. Forty-seven samples have taken from cases with plasmacytoma (Pm) (33 bones and 14 soft tissue plasmacytomas) were used as study material. Pm was the initial presentation in all cases, and bone marrow (BM) involvement was detected in 19 cases with systemic evaluation. Twenty-seven of the cases were male (age range was between 19 and 81 years). Histopathologically 27 cases had mature and 20 cases had immature morphology. Immunohistochemical analysis was used to detect VEGF expression and this was scored according to the percentage of the VEGF stained cells. VEGF expression was detected in 32 cases and in eight cases this expression was strong. In 11 cases expression was moderate and 13 cases showed mild expression. When we compared VEGF expression with pathologic grade, 17 of 20 immature samples showed VEGF expression while 15 of 27 mature samples showed VEGF expression. There was a statistically significant association between immature morphology and VEGF expression (p < 0.0264). Additionally all the samples, except one, with strong VEGF expression showed immature morphology. In conclusion two thirds of the cases with Pm showed VEGF expression and this was associated with immature morphology. Increased expression of VEGF was seen in plasmacytomas, and additional studies are needed to determine whether this translates to increased microvasculature or increased risk of progression to myeloma. PMID- 11908719 TI - An unusual presentation of plasma cell dyscrasias: cardiac tamponade due to myelomatous infiltration. AB - Pericardial involvement, a rare complication of multiple myeloma (MM), is caused by amyloidosis, infections, bleeding abnormalities or plasma cell infiltration, usually at a late or terminal stage of the disease. Three cases of MM with pericardial involvement are reported here and discussed in the light of current literature. In a retrospective review of all patients with MM at two institutions, three cases of pericardial involvement were identified. In one case, we were able to obtain cytospin preparations of the pericardiocentesis fluid. In the remaining two patients, the pericardial biopsy specimen was obtained via a pericardial window. All patients had progressive dyspnea and signs of pericardial tamponade. The pericardiocentesis fluid showed infiltration with plasma cells in one of the three patients, who had a progressive and fatal course. In the second patient pericardial invasion was proven by biopsy and the third was diagnosed with a plasma cell leukemia but developed a pericardial effusion demonstrated by pericardial biopsy. All these three patients died of progressive disease without any response to chemotherapy and supportive measures. In conclusion, optimal treatment for malignant involvement of the pericardium by myeloma cells has not yet been established and is often fatal. PMID- 11908720 TI - Level of CD 20-expression and efficacy of rituximab treatment in patients with resistant or relapsing B-cell prolymphocytic leukemia and B-cell chronic lymphocytic leukemia. AB - Rituximab (IDEC C2B8) is a chimeric human-mouse monoclonal anti-CD20 antibody that has been proven to be effective for the treatment of patients with CD20 positive leukemia and lymphoma. The level of CD20-expression in patients with B cell chronic lymphocytic leukemia (B-CLL) is low in comparison to other B-cell lymphomas and normal B-cells. Previous experience with rituximab treatment in small series of patients with B-CLL suggest that it is less effective in B-CLL than in follicular lymphomas. We analyzed the correlation between CD20-expression level and efficacy of rituximab treatment in eight patients with refractory or relapsed B-CLL and two patients with B-cell prolymphocytic leukemia (B-PLL). We could not identify any correlation between CD20-expression and efficacy of rituximab treatment. PMID- 11908721 TI - Expression of the multidrug resistance-associated protein (mrp) gene in chronic lymphocytic leukemia. AB - In order to define more accurately the role of multidrug resistance (MDR)-related protein (mrp) gene in chronic lymphocytic leukemia (CLL), we addressed the question of its expression pattern in isolated peripheral blood B lymphocytes in seven healthy donors and 28 patients with CLL, with respect to some laboratory and clinical parameters of the disease. For this purpose, we used a semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), based on the coamplification of an internal standard not homologous to the DNA target to quantify the mrp transcription level in each studied sample. We report that the level of constitutive mrp gene's expression in peripheral blood lymphocytes is higher in CLL patients than in healthy controls. We found increased mrp gene expression levels in patients with higher white blood cells (WBC) and lymphocytes' counts as well as in more advanced disease stages according to Rai or Binet scale. Finally, mrp gene's expression was higher in patients with progressive CLL, especially in cases refractory to chemotherapy salvage. The results of the present study suggest that expression of mrp gene might be relevant in the pathogenesis of the MDR phenotype in CLL. PMID- 11908722 TI - VH gene analysis in sporadic Burkitt's lymphoma: somatic mutation and intraclonal diversity with special reference to the tumor cells involving germinal center. AB - We analyzed the immunoglobulin heavy chain variable region (V(H)) gene in seven cases of sporadic Burkitt's lymphoma (sBL) to elucidate their cell of origin. In particular, we focused on the V(H) gene status of tumor cells involving adjacent germinal center (GC) by microdissecting histological sections. Among the seven V(H) genes V(H)1 family was found in two, V(H)3 in four, and V(H)4 in one. All rearranged V(H) genes demonstrated somatic mutations at percentages ranging from 1.4 to 7.5% (mean, 4.2%), which is a similar level to that seen in IgM-only B cells. Three out of four V(H) genes with more than 2% sequence difference from their corresponding germline counterpart showed evidence of antigen selection in their framework region 3. Three cases demonstrated signs of intraclonal diversity with a mutational frequency of 0.47-0.98%, which was 13.5-28.8 times as great as the Taq infidelity in our experimental conditions. However the level of somatic mutation and the effect of antigen selection on V(H) gene were diverse in these three cases, and the relationship between V(H) gene somatic mutation status and intraclonal diversity was unclear in sBL. In the analysis of microdissected tissues, all 20 tumor clones in the adjacent GCs showed additional replacement mutations in complementarity determining region 3, suggesting a role of antigen in tumor progression. This finding resembles the phenomenon that memory B-cells reenter into GC to undergo further affinity maturation. In contrast, 7/11 V(H) gene sequences irrelevant to GC were identical to those of the major tumor clone. Thus our findings suggested that sBL is derived from memory B-cells rather than GC B-cells, and that antigen stimulation is involved in the clonal expansion of a proportion of sBL. PMID- 11908723 TI - Characterization of a novel human anaplastic large cell lymphoma cell line tumorigenic in SCID mice. AB - L82, a novel anaplastic large cell lymphoma (ALCL) cell line was established from the pleural effusion of a 24-year-old patient with recurrent ALCL. L82 cells showed the typical morphologic features of ALCL cells with irregular, often indented, nuclear profiles, prominent nucleoli, and abundant cytoplasm. The immunoprofile of L82 corresponds to that seen typically in primary ALCL cells, with positivity for CD30, EMA, CD3, CD4, CD25, CD71, TIA1, and granzyme B; the cells were negative for EBV-related antigens. Cytogenetic analysis showed a complex, near triploid karyotype with 72-77 chromosomes, including the ALCL specific translocation t(2;5)(p23;q35). Chromosomal analysis revealed a number of secondary structural alterations including amplification of 7q21-31, 1q, and 6p, and gain of chromosomal material in 8q (affecting the c-myc gene). The rearrangement of the T-cell receptor-gamma locus shows that L82 is clonally derived from T-lineage lymphoid cells. mRNAs for interleukin 7 (IL-7), IL-8, IL 9, IL-10, TNF-beta, and for the IL-7 and IL-9 receptor were found. These data show that the T-helper cell (Th)1/Th2 balance was polarized to Th2. L82 were inoculated intraperitoneally into 4 week-old SCID mice and produced a disseminated tumor within 4-6 weeks. Morphological, immunohistochemical, and molecular genetic investigation confirmed that the xenograft and the original ALCL tumor were identical. SCID mice xenografted with the human ALCL cell line, L82, provide a useful model system for the investigation of the biology of ALCL and of new therapeutic approaches, such as specific immunotherapy. PMID- 11908724 TI - Quantitative analysis of cell-free Epstein-Barr virus genome copy number in patients with EBV-associated hemophagocytic lymphohistiocytosis. AB - To determine whether the EBV genome content in serum or plasma reflects clinical features and outcome in EBV-associated hemophagocytic lymphohistiocytosis (EBV HLH), we quantified the cell-free EBV genome copy number by real-time PCR in 38 patients with EBV-HLH, and compared this to the values from 15 patients with infectious mononucleosis (IM). The median (range) cell-free EBV genome copy number at diagnosis was 3.0 x 10(3) (undetectable -5.5 x 10(7)) copies/ml in EBV HLH, which was significantly higher than the 6.6 x 10(1) (undetectable -1.0 x 10(3)) copies/ml in IM (P = 0.0008). We serially analyzed cell-free EBV genome copy number in 10 cases of EBV-HLH up to 4 months from diagnosis. In four patients who achieved remission, the EBV genome became undetectable soon after starting therapy. In the remaining six patients who responded poorly to therapy, the EBV genome copy number in the serum or plasma remained at high levels except for one case. In addition, we confirmed that the EBV genome became undetectable after hematopoietic stem cell transplantation in 4 EBV-HLH cases. These results suggest that the quantitative analysis of cell-free EBV genome copy number is useful for evaluating disease activity and for predicting the response to therapy in EBV-HLH. PMID- 11908725 TI - Malignant evolution of Schnitzler's syndrome--chronic urticaria and IgM monoclonal gammopathy: report of a new case and review of the literature. AB - Schnitzler's syndrome, initially described in 1974 is an uncommon condition defined by chronic urticaria and monoclonal IgM gammopathy. Additional features include fever of unknown origin, elevated ESR, bone pain and frequently a benign clinical course. We conducted a literature search of Medline, EMBASE and Cancerlit and found 56 cases of Schnitzler's syndrome reported to date. The absence of lymphoproliferative disease in this condition is typical, but nine patients have progressed to develop lymphoplasmacytic neoplasias, particularly Waldenstrom's macroglobulinemia (WM). Malignant evolution of Schnitzler's syndrome is a rare complication, but emphasizes the importance of long term follow-up and the need for these patients to undergo periodic assessment of the bone marrow and lymph nodes. Treatment of this condition is difficult, with varying response to corticosteroids and largely unsuccessful results with standard chemotherapy used for WM. We describe a case of Schnitzler's syndrome in a 50-year old man with lymphocytic aggregates in the bone marrow after 9 years of chronic urticaria, fever, arthralgias and bone pain. We review the clinical features and treatment, with emphasis on the hematologic aspects of this unusual condition. PMID- 11908726 TI - Bladder involvement of diffuse large B-cell lymphoma diagnosed by a cytological study of the urine. AB - The diagnosis of lymphoma involvement from the urine sediment has been rarely reported in the literature. We present a 78-year-old woman with diffuse large B cell lymphoma in whom a relapse of the disease was diagnosed due to the presence of lymphoma cells in the urine. The analysis of urine by flow cytometry demonstrated a clonal B-cell origin, with an identical immunophenotype to that of the initial lymphoma. This case emphasizes the interest of cytological studies of the urine in cases with suspected bladder involvement by lymphoma. PMID- 11908727 TI - Survival of B lineage leukemic cells: signals from the bone marrow microenvironment. AB - Stromal cells are an essential component of the bone marrow microenvironment that regulate development of immature hematopoietic progenitor cells. Through production of soluble cytokines, and signaling through adhesion molecule interactions, stromal cells impact survival, proliferation, and differentiation of hematopoietic progenitor cells. Similarities between normal pro-B and pre-B cells and B lineage acute lymphoblastic leukemic (ALL) progenitors have been well characterized which provide a model for investigation of the mechanisms by which ALL cells respond to bone marrow microenvironment signals. In addition to providing survival signals to B lineage ALL during initiation of disease, the bone marrow has long been recognized as a "sanctuary site" for leukemic cells during traditional chemotherapy. In the current review, mechanisms by which stromal cells contribute to leukemic cell survival, and the potential impact on treatment efficacy, are discussed. A growing appreciation of the significance of the bone marrow microenvironment in the progression of ALL, and further investigation of the signaling between leukemic progenitors and stromal cells, may contribute to novel treatment strategies aimed at enhancing sensitivity of ALL cells to currently available chemotherapeutic agents. PMID- 11908728 TI - Simultaneous occurrence of myelodysplastic syndrome and monoclonal B lymphocytes with a different clonal origin. AB - Bone marrow and peripheral blood from a myelodysplastic syndrome patient with trisomy 13 and monoclonal B lymphocytes (without evidence of systemic lymphoma) were investigated for clonal lymphoid lineage involvement using interphase fluorescence in situ hybridization (FISH) and X-chromosome inactivation assay (HUMARA) on CD19+ and CD34+ sorted cells. Trisomy 13 was detected in 55% of CD34+ cells and in 5.5% of CD19+ cells, the latter being comparable to the negative control specimen. X-chromosome inactivation showed both CD34+ and CD19+ cells to be monoclonal, though their inactivated X-chromosome was different. The results strongly suggested that both populations of CD34+ and CD19+ cells have originated from a different progenitor stem cell. PMID- 11908729 TI - Immunoglobulin and T-cell receptor gene-gene rearrangement in pleural cavity based T-cell rich B-cell lymphoma in an immunocompetent patient. AB - Body cavity-based lymphomas are fluid-based lymphomas that are not associated with a tumor mass or adenopathy which could explain the origin of the lymphomatous effusion. A distinct lymphoma that grows in the body cavity as a lymphomatous effusion in the absence of a tumor mass has been identified as a primary effusion lymphoma. This almost exclusively occurs in patients with acquired immunodeficiency syndrome (AIDS), who invariably have a history of Kaposi sarcoma. We report a rare case of a recurrent pleural effusion in an immunocompetent patient. There was no evidence of lymphadenopathy or an associated mass on computerized tomography of the chest, abdomen and pelvis. Serology for HIV, HHS-8, EBV and HTLV-1 were negative. Cytologic examination of the pleural fluid showed an elevated white cell count with 97% lymphocytes, mostly with T-cell markers. Bone marrow aspirate and biopsy were negative and bronchoscopy was unrevealing. Pleural biopsy was significant for >70% T lymphocytes and some large atypical cells. Which had CD19, CD20 and weak bcl-2 positivity. Kappa and lambda light chains did not show distinct clonality. A preliminary diagnosis of T-cell rich B-cell lymphoma (TCRBCL) of the pleural cavity was made. The diagnosis was confirmed with DNA studies done on the pleural biopsy specimen using PCR and southern blot. Dual rearrangement of Ig heavy chain region and TCR-beta genes were identified. The patient responded to combination chemotherapy with cyclophosphamide, adriamycin, vincristine and prednisone. Our case is the first known case of pleural cavity-based TCRBCL and illustrates the role of gene rearrangement studies in such patients. PMID- 11908730 TI - Leukemic leptomeningeal involvement in stage 0 and stage 1 chronic lymphocytic leukemia. AB - Central nervous system (CNS) involvement in early (Rai Stage 0 and Stage 1) chronic lymphocytic leukemia (CLL) is rare, with only five cases reported. We present the sixth reported case, a 77-year-old male with a 4 year history of Stage 0 CLL who presented with sudden onset of diplopia and headache. Workup revealed a leukemic involvement of his CNS and he responded well to treatment with intrathecal (IT) methotrexate. After his third IT treatment, he developed a change in his mental status, consistent with a chemotherapy induced encephalopathy, which was effectively treated with IT hydrocortisone. In addition to the case presentation, we review the previously reported cases in an effort to determine any characteristics common among the Stage 0/1 CLL patients with reported CNS involvement. PMID- 11908731 TI - Nasal NK-cell lymphoma followed by relapse in the uterine cervix. AB - We report a case of nasal natural killer (NK)-cell lymphoma in a 51-year-old Japanese woman who showed a later relapse in the uterine cervix. The nasal NK cell lymphoma regressed after local radiation therapy. Six months after the diagnosis while the patient was being treated with chemotherapy for a subclinical tumor, a mass lesion of the uterine cervix was noticed by follow-up computed tomography. Giemsa-stained vaginal smear showed lymphoid tumor cells with large azurophilic granules, leading to a rapid diagnosis of cervical involvement by NK cell lymphoma. The chemotherapy regimens were immediately changed, but the patient died 2 months after the relapse with an overall survival of 8 months. This case may be of value in elucidating the biological behavior and natural history of NK-cell lymphoma. PMID- 11908732 TI - Pentasomy 8q resulting from duplication of isochromosome 8q in chronic myelomonocytic leukemia. AB - We report an interesting case of chronic myelomonocytic leukemia (CMML) with pentasomy 8q resulting from the duplication of isochromosome 8q in a 47-year-old male. His blood picture and myelogram showed CMML and the chromosome study, using R-banding and G-banding techniques, revealed a karyotype of 47,XY, 8,+i(8)(q10)x2. Dual-color fluorescence in situ hybridization (FISH) studies with a #8 centromeric probe and a locus-specific probe for C-myc gene completely confirmed the result of the conventional cytogenetic method. Reverse transcription polymerase reaction (RT-PCR) revealed no BCR/ABL fusion transcript. Hydroxyurea and 6-mercaptopurine therapy did not induce a complete remission and five months later he died of exacerbation of his disease. On reviewing another two cases with pentasomy 8q in the literature, we feel that pentasomy 8q, when present as a sole anomaly, may play a specific role in leukemogenesis and in determining the clinical characteristics such as monocytic involvement and poor prognosis. PMID- 11908733 TI - Idiopathic eosinophilic endomyocarditis in the absence of peripheral eosinophilia. AB - In this case report we present two patients with unusual manifestations of eosinophilic endomyocarditis: A 69-year-old patient with a history of heart failure and ventricular fibrillation and a 16-year-old woman with ventricular fibrillation and an ECG indicating acute myocardial infarction had both normal blood eosinophil counts at the onset of symptoms. The absence of hypereosinophilia, therefore, does not exclude the presence of eosinophilic organ infiltration. Endomyocardial biopsy may be the only diagnostic procedure to identify necrotic eosinophilic endomyocarditis in patients with unexplained heart failure or ventricular fibrillation. PMID- 11908734 TI - Ikaros gene expression and leukemia. AB - The Ikaros (Ik) protein, or LyF1, was initially described as a protein binding to regulatory sequences of a number of genes expressed in murine lymphoid cells. Ikaros is a critical regulator of normal hematopoietic stem cell differentiation, as evidenced by dramatic defects in the lymphoid compartments, in homozygous animals with gene inactivation. Because differential splicing produces multiple isoforms with potentially different functions, Ikaros provides a unique model to study how post-transcriptional mechanisms may be involved in neoplastic processes. Indeed, several groups including ours have underlined evidences that expression of different Ikaros isoforms vary among different types of leukemias. The predominance of short isoforms in certain subsets is intriguing. Here, additional observations reinforced the hypothesis that Ikaros expression may be deregulated in human leukemias. Whether this is a cause or a consequence of the leukemic process remains speculative. Other human diseases however, provide examples of abnormal post-transcriptional regulations that have been further characterized. PMID- 11908735 TI - Adenovirus infection of primary malignant lymphoid cells. AB - Adenovirus infection represents a cellular stress that induces host cell pro apoptotic responses. To overcome this barrier to productive infection, viral polypeptides modulate a variety of host cell pathways. The interface of these early viral gene products with key cellular regulatory proteins has provided considerable information concerning basic cellular mechanisms operative in cell cycle regulation, transcriptional control and apoptosis. The overlap of these mechanisms with those impacted during oncogenesis provides the opportunity to use adenoviruses and adenovirus mutants to characterize the state of key regulatory pathways in specific malignant cells. For example, adenoviruses mediate cytotoxicity after infection of chronic lymphocytic leukemia (CLL) cells, mantle cell lymphoma (MCL) cells and multiple myeloma cell lines. Specific adenovirus mutants demonstrate enhanced cytotoxicity and, in many cases, apoptosis is not the primary mechanism of cell death. Analysis of these infections with respect to both the features of the primary malignant cell and the mechanisms of adenovirus mediated cytotoxicity holds the prospect of providing novel insights into the status of key regulatory pathways in individual patient malignant cells. These studies also hold the prospect of supporting the development of specific attenuated adenoviruses as therapeutic agents with selective cytotoxicity for specific primary lymphoid malignancies. PMID- 11908737 TI - Rearrangements of chromosome band 3q21 in myeloid leukemia. AB - Chromosome rearrangements affecting band 3q21, namely, the inv(3)(q21q26), the t(3;3)(q21;q26), and the t(1;3)(p36;q21), are associated with a particularly poor prognosis in myeloid leukemia or myelodysplasia. Originally, inv(3) and t(3;3) breakpoints have been reported to cluster in a region (breakpoint cluster region, BCR) of approximately 30 kb, which is located centromeric and downstream of the ribophorin I (RPN-I) gene. More recently, we established a PAC contig that includes the 3q21 BCR, and used these PAC clones to map breakpoints in patient samples by both metaphase and interphase fluorescence in situ hybridization (FISH) analysis. A significant proportion of inv(3) and t(3;3) breakpoints was located at sometimes considerable distances centromeric of the originally described BCR, in a region recently also implicated in t(1;3) rearrangements. These breakpoints may thus define a second, centromeric BCR (BCR-C), or extend the original 3q21 BCR to a size of approximately 100 kb. Activation of the EVI-1 gene in 3q26 by regulatory sequences of the housekeeping gene RPN-I has been suggested as a leukemogenic mechanism in patients with inv(3) and t(3;3). However, despite a number of characteristics that make EVI-1 an attractive candidate oncogene, its biological properties fail to fully explain the phenotype of leukemias carrying 3q rearrangements. Several additional candidate genes have been identified in or near the 3q21 breakpoint region, but their possible contribution to the characteristics of leukemias with 3q21 rearrangements remains to be explored. PMID- 11908736 TI - Survival and cell cycle control in early hematopoiesis: role of bcl-2, and the cyclin dependent kinase inhibitors P27 and P21. AB - Homeostasis of the hematopoietic system is maintained by proliferation and differentiation of a small number of long-term surviving, self-renewing stem cells, which give rise to the fully mature elements. The fine interplay between differentiation, proliferation and death by apoptosis determines the equilibrium of this system. Thus, genes involved in the control of these processes are very important in the regulation and development of hematopoietic cells especially in the initial stages. The interactions among cyclins, their specific cyclin dependent kinases (CDKs) and, a number of cyclin-dependent kinase inhibitors (CDKIs) such as p27 and p21, exert a direct control on the cell cycle but can also produce other independent effects on hematopoietic differentiation. Proteins of the Bcl-2 family are also crucial in regulating the balance between entry into apoptosis and survival capacity and their roles change in the course of differentiation. In addition, a number of autocrine and paracrine soluble factors (such as TGF-beta1) modulate the behavior and differentiation potential of hematopoietic elements. Studies on a few in vitro systems of early hematopoietic differentiation have stressed the importance of Bcl-2 and of the CDKIs p27 and p21 at this stage, have confirmed cell-cycle independent effects and have demonstrated how the modulation and the effects in response to different stimuli is mostly dependent on the differentiation stage of the target cells. PMID- 11908738 TI - Myeloablative treatments for multiple myeloma: update of a comparative study of different regimens used in patients from the Spanish registry for transplantation in multiple myeloma. AB - After a previous analysis that did not detect clear differences in the results of three conditioning regimens used for autologous stem cell transplantation (ASCT) in patients from the Spanish Registry for Transplant in Multiple Myeloma (MM), we have updated the registry, including a larger number of cases and a fourth conditioning regimen with a longer follow-up. We evaluate 472 MM patients treated with 200 mg/m2 melphalan (MEL200), 135 patients treated with 140 mg/m2 melphalan plus total body irradiation [(MEL140 + TBI)], 186 patients treated with 12 mg/kg busulphan plus 140 mg/m2 melphalan (BUMEL) and 28 patients treated with 14 mg/kg busulphan followed by cyclophosphamide 120 mg/kg (BUCY). There were no significant differences in respect to either transplant related death or haematological recovery, regardless of growth factor use, between the four conditioning programs. Nevertheless, hospitalization time with MEL200 was less than with BUMEL when growth factors were used (19+/-9 vs. 25+/-9 days, P = 0.009) and less than with MEL140 + TBI without growth factors (20+/-8 days vs. 27+/-9 days, P = 0.002). In patients with measurable disease at ASCT (non-complete remission [CR]), BUMEL achieved a 51% CR vs. 43%-31% in the other groups (P = 0.007). The response rate for patients in partial remission (PR) at ASCT was 100% with BUMEL vs. 93%-86% in the other groups (P between 0.02 and 0.0007). The median overall survival (OS) for the BUMEL group was 57 months (95% confidence interval [CI]: 51-78) as compared to 45 (CI: 36-64) months for the MEL200 group and 39 (CI: 28-72) months for the MEL140 + TBI and BUCY groups. The median event free survival (EFS) was longer in the BUMEL group [30 (CI: 22-44) mo] than in the MEL200 [22 (CI: 18-26) mo], BUCY [23 (CI: 11-50) mo] or MEL140 + TBI groups [20 (CI: 15-29) mo]. Nevertheless, the differences in OS and EFS did not reach statistical significance in either the univariate analysis or the multivariate analysis adjusted with other high prognostic weight factors. As in the initial study, differences in regards to the anti-myeloma effect of the conditioning regimens are not conclusive. However, the better response rates associated with the favorable tendency in outcome achieved with BUMEL, continue to justify further prospective studies. PMID- 11908740 TI - Prognostic factors in elderly patients with acute myelogenous leukemia: a single center study in Japan. AB - We retrospectively analyzed data of 47 patients aged 60 years or older, hospitalized in our institution with the diagnosis of acute myelogenous leukemia (AML), and searched for prognostic factors. Induction with anthracyclines significantly correlated with better complete remission (CR) rate (P = 0.0016) and overall survival (OS) (P < 0.001). Another factor significantly affecting CR rate was higher age (> 70 years) (P = 0.042). Therapy-non-related factors predictive for shorter OS in univariate analyses were age older than 70 years (P = 0.003), percentage of blasts in bone marrow more than 80% (P = 0.048), serum lactate dehydrogenase level higher than 250 U l(-1) (P = 0.032). In stepwise cox proportional hazard regression model, all the four factors predictive for poor OS remained to be independently and significantly prognostic for shorter OS. Only two patients receiving anthracyclines died within 30 days and the frequency was not different from that in patients not receiving anthracyclines. The use of anthracyclines as induction therapy is recommended even in the elderly patients. PMID- 11908739 TI - Treatment of acute lymphoblastic leukemia in the elderly: an evaluation of interferon alpha given as a single agent after complete remission. AB - Although interferon (IFN) has been used in elderly patients with acute lymphoblastic leukemia (ALL), the benefits from IFN therapy have not been properly assessed, especially as it was given combined with other cytotoxic drugs, which obscured the role of IFN if any. In 1997, we started a study aimed at improving our previous results in elderly patients with ALL and at assessing the therapeutic role of IFN in this disease. Fifty-eight patients with ALL, aged 55-81 years (median: 64.9 years), were randomly allocated to treatment with vindesine or vincristine during induction. After a first consolidation course, IFN was administered as a single agent for three months together with cranial radiotherapy. Chemotherapy was then resumed with a second consolidation course and maintenance. A complete remission (CR) was obtained in 58% of patients (CI: 45-71%), significantly less than in our previous study which included IFN combined with chemotherapy during maintenance (CR: 85%, CI:70-94%, p = 0.007). Overall survival (median: 289 vs 434 days in the previous study, p = 0.01) and disease-free survival (median: 146 vs 427 days, p = 0.009) were also inferior in the present study. In particular, the pattern of relapses over time suggested that the 3 month IFN treatment phase with no additional chemotherapy might have contributed to the comparatively poor outcome of this cohort. In addition, vindesine given during induction did not prove less neurotoxic than vincristine, did not improve the CR rate, and had no impact on survival. In conclusion, although similar to published studies in elderly patients with ALL, this study is inferior to our previous one. INF, given as a single drug, has a modest role if any in the treatment of older persons with ALL. PMID- 11908741 TI - Unrelated donor leukocyte infusions to treat relapse after unrelated donor bone marrow transplantation. AB - Allogeneic stem cell transplantation (SCT) may cure many patients with hematologic malignancies due to both the intensive conditioning therapy, and in many patients, the potent graft-vs-leukemia (GVL) effect of the donor graft. The GVL effect is mediated in large part by mature T-cells contained in the donor graft and has been defined in detail in animal models of transplantation. The GVL activity has been observed in the clinical setting after SCT from both matched siblings and unrelated donors. The best demonstration and most direct evidence of GVL activity in humans come from the use of donor leukocyte infusions (DLI). For patients who relapse with chronic myelogenous leukemia after matched sibling SCT, infusions of leukocytes collected from the original transplant donor will re establish complete and durable remission in 60-80% of patients. DLI is less effective for more advanced phases of CML and for patients who relapse with diseases other than CML. DLI after matched sibling SCT is complicated primarily by graft-vs-host disease (GVHD), marrow aplasia, and unfortunately, relapse in some cases. There has been little information regarding the use of unrelated DLI (UDLI). Available data now shows that despite initial concerns that UDLI would result in excessive toxicity, it is an effective approach to relapse after unrelated donor marrow grafting. Response rates are similar to those seen after the use of matched sibling DLI, and many remissions remain durable. Graft-vs-host disease is a frequent complication after UDLI though the incidence and severity of GVHD is also similar to the use of matched sibling DLI. It is not clear that the GVL and GVHD effects can be separated, since the majority of responding patients also develop GVHD. The most effective cell dose for UDLI has not been established, though there does not appear to be either a dose-response or dose toxicity relationship from UDLI. Although second unrelated donor bone marrow transplantation (BMT) may cure a small minority of patients, GVL induction with UDLI offers a safer and potentially more effective therapy for relapsed leukemia, and offers insights in methods to manipulate the human immune system for therapeutic benefit. PMID- 11908742 TI - Diffuse large B-cell lymphoma: a clinicopathologic analysis of 444 cases classified according to the updated Kiel classification. AB - The purpose of this study was: to compare the survival of diffuse large B-cell lymphomas (DLBCL) stratified according to the up-dated Kiel classification. A retrospective study of a cohort of 1378 cases was organized in 1996 by the Non Hodgkin's Lymphoma Classification Project, and the DLBCL were classified according to the updated Kiel classification. The distribution of the different types and subtypes was as follows: centroblastic (CB, 85.4%), composed of the polymorphic (CB-PM, 58.6%), monomorphic (CB-MM, 17.1%) and multilobated (CB-ML, 9.7%) subtypes; immunoblastic (IB, 11.2%), with (8.3%) or without (2.9%) plasmacytoid differentiation; and anaplastic large cell lymphoma (ALCL) of B-cell type (3.4%). The rate of diagnostic agreement between pathologists was 78% for CB and 65% for IB lymphoma. The 5-year overall survival (OAS) for the entire group was 47% and the 5-year failure-free survival (FFS) was 42%. No significant differences in survival were found between the three major groups (CB, IB, ALCL). However, the 5-year OAS and FFS of patients with DLBCL not containing immunoblasts (CB-MM+CB-ML) was 51 and 52%, respectively, and was significantly better than the survival of those containing immunoblasts (CB-PM+IB+ALCL), which was 44 and 38% (p = 0.06 and p = 0.037), respectively. These results did not appear to be due to differences in the clinical features of the two groups, and was most significant for patients with low stage or low risk disease. However, histologic subtyping was not an independent risk factor for the entire group by multivariate analysis. In conclusion, patients with CB-MM and CB-ML (without immunoblasts) had a significantly better OAS and FFS than those with CB-PM, IB and ALCL (with immunoblasts). Therefore, we conclude that additional studies are still needed to further evaluate the importance of immunoblastic differentiation in DLBCL. PMID- 11908743 TI - Zinc nutrition and HIV infection. AB - The trace element zinc is involved in many important immune processes. A number of immunologic impairments owing to zinc deficiency are also evident in HIV disease, most notably a reduction in the number of circulating T lymphocytes. Observational epidemiologic studies have provided conflicting results on the role of zinc status in HIV disease progression. Randomized, placebo-controlled trials are needed to resolve this controversy. Studies must also address the role of zinc in vertical transmission of HIV from mother to child and its role in reducing the risk of adverse pregnancy outcomes, both of which are of considerable public health importance in developing countries. PMID- 11908744 TI - Involvement of inositol in reproduction. AB - Inositol is involved in several aspects of reproduction. It affects overall embryogenesis, may prevent neural tube defects, and stimulates the production of lung surfactant. This article will review the involvement of inositol in reproduction. After describing the biologic function of inositol and its derivatives, studies are quoted in which the role of inositol in fertility, embryogenesis, fetal development, and pregnancy outcome are examined. PMID- 11908745 TI - The effect of low and high doses of beta-carotene and exposure to cigarette smoke on the lungs of ferrets. AB - When the diets of ferrets were supplemented with large (pharmacologic) daily doses of beta-carotene (BC) for 6 months, the levels of retinoic acid and the retinoic acid receptor beta declined significantly in lung tissues. Indicators of cell proliferation (c-jun and c-fos proteins and others) increased. Histologic observations showed that feeding high doses of BC resulted in keratinized squamous metaplasia in the lung tissues. When high-doses of BC were combined with daily exposure to cigarette smoke, the BC effects were greatly accentuated. These results may lead to an explanation of the increased incidence of lung cancer in two large independent epidemiologic studies of smokers in which pharmacologic doses of BC were given. PMID- 11908746 TI - All that glitters is not iron (deficiency): revisiting the question of why anemic individuals are anemic. AB - Three recent surveys from Kenya, the Ivory Coast, and East Java emphasize that anemia is a complex and multifaceted public health problem. A thorough understanding of iron metabolism and its disturbance is central to resolving the problem of anemia but requires intricate analysis. To maximize oxygen-carrying capacity and optimize human function, one must combine issues of sociology, economics, and general health with multiple micronutrient interventions. PMID- 11908747 TI - Case-control association studies in pharmacogenetics. PMID- 11908748 TI - Chipping away at the mystery of drug responses. PMID- 11908749 TI - Jets: a modification to speed flexible oligonucleotide array construction. PMID- 11908750 TI - Oxidative stress and redox imbalance in acetaminophen toxicity. PMID- 11908752 TI - Pharmacogenomic testing: the cost factor. PMID- 11908751 TI - Integrating genotype and phenotype information: an overview of the PharmGKB project. Pharmacogenetics Research Network and Knowledge Base. PMID- 11908753 TI - The pharmacogenomics of depression. PMID- 11908754 TI - Recent advances in P450 research. AB - P450 enzymes comprise a superfamily of heme-containing proteins that catalyze oxidative metabolism of structurally diverse chemicals. Over the past few years, there has been significant progress in P450 research on many fronts and the information gained is currently being applied to both drug development and clinical practice. Recently, a major accomplishment occurred when the structure of a mammalian P450 was determined by crystallography. Results from these studies will have a major impact on understanding structure-activity relationships of P450 enzymes and promote prediction of drug interactions. In addition, new technologies have facilitated the identification of several new P450 alleles. This information will profoundly affect our understanding of the causes attributed to interindividual variations in drug responses and link these differences to efficacy or toxicity of many therapeutic agents. Finally, the recent accomplishments towards constructing P450 null animals have afforded determination of the role of these enzymes in toxicity. Moreover, advances have been made towards the construction of humanized transgenic animals and plants. Overall, the outcome of recent developments in the P450 arena will be safer and more efficient drug therapies. PMID- 11908755 TI - Mouse anxiety: the power of knockout. AB - The role of the neurokinine 1 receptors (NK1R) and its endogenous ligand, the neuropeptide substance P (SP) in the pathophysiology and treatment response of anxiety and depression has been controversial. In a new study, however, mice with a targeted inactivation of the NK1R show a phenotype that is associated with anxiety-related behaviors and stress responses. Since this behavioral phenotype was associated with an increase in serotonergic function, NK1R knockout mice represent a systematic model for advanced investigations of SP/NK1R and serotonin system interaction in anxiety responses. The availability of an increasing number of knockout mouse anxiety models as well as integration of emerging tools and technologies for genetic analysis will provide the groundwork for the genetic dissection of anxiety and related disorders. PMID- 11908756 TI - Single nucleotide polymorphism identification in candidate gene systems of obesity. AB - We have constructed a large panel of single nucleotide polymorphisms (SNP) identified in 68 candidate genes for obesity. Our panel combines novel SNP identification methods based on EST data, with public SNP data from largescale genomic sequencing, to produce a total of 218 SNPs in the coding regions of obesity candidate genes, 178 SNPs in untranslated regions, and over 1000 intronic SNPs. These include new non-conservative amino acid changes in thyroid receptor beta, esterase D, acid phosphatase 1. Our data show evidence of negative selection among these polymorphisms implying functional impacts of the non conservative mutations. Comparison of overlap between SNPs identified independently from EST data vs genomic sequencing indicate that together they may constitute about one half of the actual total number of amino acid polymorphisms in these genes that are common in the human population (defined here as a population allele frequency above 5%). We have analyzed our polymorphism panel to construct a database of detailed information about their location in the gene structure and effect on protein coding, available on the web at http://www.bioinformat ics.ucla.edu/snp/obesity. We believe this panel can serve as a valuable new resource for genetic and pharmacogenomic studies of the causes of obesity. PMID- 11908757 TI - The predictive value of MDR1, CYP2C9, and CYP2C19 polymorphisms for phenytoin plasma levels. AB - Phenytoin, an anticonvulsant, exhibits nonlinear pharmacokinetics with large interindividual differences. Because of its small therapeutic range with the risk of therapeutic failure or adverse drug effects in susceptible persons, therapeutic drug monitoring is frequently applied. The interindividual differences in dose response can partially be explained by known genetic polymorphisms in the metabolic enzyme CYP2C9 but a large deal of individual variability remains still unexplained. Part of this variability might be accounted for by variable uptake of phenytoin, which is a substrate of p glycoprotein, encoded by the human MDR1 gene. We evaluated, whether phenytoin plasma levels correlate with a polymorphism in the MDR1 gene, C3435T, which is associated with intestinal PGP activity. Genotyping and analyses of plasma levels of phenytoin and metabolites in 96 healthy Turkish volunteers showed that the MDR1C > T3435 polymorphism affects phenytoin plasma levels (P = 0.064) and the metabolic ratio of p-HPPH vs phenytoin (MDR1*TT genotype, P = 0.026). The MDR1*CC genotype is more common in volunteers with low phenytoin levels (P < or = 0.001, chi2 test). A combined analysis of variable alleles of CYP2C9, 2C19 and MDR1 revealed that the number of mutant CYP2C9 alleles is a major determinant, the number of MDR1*T alleles further contributes to the prediction of phenytoin plasma levels and CYP2C19*2 does not explain individual variability. The regression equation that fitted the data best included the number of mutant CYP2C9 and MDR*T alleles as predictory variables and explained 15.4% of the variability of phenytoin data (r2 = 0.154, P = 0.0002). Furthermore, analysis of CYP2C9 and MDR1 genotypes in 35 phenytoin-treated patients recruited from therapeutic drug monitoring showed that combined CYP2C9 and MDR1 analysis has some predictive value not only in the controlled settings of a clinical trial, but also in the daily clinical practice. PMID- 11908758 TI - Genomic analysis of a mouse model of immunoglobulin A nephropathy reveals an enhanced PDGF-EDG5 cascade. AB - The molecular mechanism of immunoglobulin A nephropathy (IgAN), the most common primary renal glomerular disease worldwide, is unknown. HIGA (high serum IgA) mouse is a valid model of IgAN showing almost all of the pathological features, including mesangial cell proliferation. Here we elucidate a pattern of gene expression associated with IgAN by analyzing the diseased kidneys on cDNA microarrays. In particular, we showed an enhanced expression of several genes regulating the cell cycle and proliferation, including growth factors and their receptors, as well as endothelial differentiation gene-5 (EDG5), a receptor for sphingosine 1-phosphate (SPP). One of the growth factors, platelet-derived growth factor (PDGF) induces a marked upregulation of EDG5 in proliferative mesangial cells, and promotes cell proliferation synergistically with SPP. The genomic approach allows us to identify families of genes involved in a process, and can indicate that enhanced PDGF-EDG5 signaling plays an important role in the progression of IgAN. PMID- 11908759 TI - The acetylcholine-binding protein: 'What's in a name?'. PMID- 11908760 TI - Regulation of drug transport by new xenobiotic receptors. PMID- 11908761 TI - How the G protein-coupled receptor activates GTP-binding protein. PMID- 11908762 TI - Combinatorial chemistry: starting the second decade. PMID- 11908763 TI - Database designs for microarray data. PMID- 11908764 TI - Multigenic traits and risk assessment in pharmacology: a population approach. PMID- 11908765 TI - When drugs development meets genomics. PMID- 11908766 TI - Pharmacogenetics: the ethical context. PMID- 11908767 TI - The pharmacogenomics of HIV therapy. PMID- 11908768 TI - Recent advances in the pharmacogenomics of thiopurine methyltransferase. AB - The thiopurine drugs (6-mercaptopurine, 6-thioguanine and azathioprine) are commonly used cytotoxic agents and immunosuppressants. One important route for the metabolism of these agents is methylation, mediated by thiopurine methyltransferase (TPMT). It is now well established that inter-individual variation in sensitivity to thiopurines can be due to the presence of common genetic polymorphisms affecting the TPMT gene. More recently variations in the number of tandem repeats in the 5' promoter region have been shown to influence TPMT expression in vitro. In this article, we review recent advances in the understanding of the range of inter-individual variation that may be involved in the open reading frame or promoter region of the TPMT gene. We also review the data which have been published regarding the influence such variations may have on both the clinical efficacy and toxicity of the thiopurine drugs. PMID- 11908769 TI - Vesicular protein transport. AB - Protein transport and sorting in the secretory and endocytic pathways via vesicles is required for organelle biogenesis, constitutive and regulated secretion and constitutive and regulated endocytosis. It is essential for a multicellular organism and the function of its specialised cell types that the multiple transport and sorting events are highly accurate. They determine the protein and lipid composition of specialised compartments, receptor protein function and membrane homeostasis. This review describes the individual events involved in the process of vesicle mediated protein transport and sorting and summarizes the knowledge about the function of proteins and lipids orchestrating the process. PMID- 11908770 TI - CYP2C9*3 influences the metabolism and the drug-interaction of candesartan in vitro. AB - Candesartan cilexetil is an angiotensin II receptor antagonist, and candesartan, its active metabolite, is metabolized by CYP2C9. However, the effect of CYP2C9*3 on candesartan metabolism is not established. We characterized the kinetics of candesartan by CYP2C9*1/*1 and CYP2C9*1/*3 in human liver microsomes. The difference between the two was not significant. Subsequently, CYP2C9*1 and CYP2C9*3 (Leu359) were expressed in yeast, and the kinetics of candesartan were determined. The wild-type showed the lower Km (345 vs 439 microM; 3/4) and higher Vmax/Km (1/3) than the Leu359 variant. Also, we investigated potential interaction between candesartan and warfarin with both the wild-type and the Leu359 variant. Candesartan had no effect on S-warfarin 7-hydroxylation. In contrast, S-warfarin inhibited candesartan metabolism by the wild-type (K = 17microM) greater than by the Leu359 variant (Ki = 36 microM). These findings suggest that CYP2C9*3 may change not only the metabolic activity but also the inhibitory susceptibility compared with CYP2C9*1. PMID- 11908771 TI - Immune-mediated heart disease: in the footsteps of Jenner. PMID- 11908772 TI - Molecular mechanisms of anti-G-protein-coupled receptor autoantibodies. PMID- 11908773 TI - Role of cytokines in autoimmune myocarditis and cardiomyopathy. AB - Cellular as well as humoral autoimmune responses are critically associated with the pathogenesis and progression of myocarditis and cardiomyopathy. Cytokines appear to play critical roles in accentuating or regulating autoimmune mechanisms in these disorders. However, depending on the triggers of autoimmune responses against the heart, such as viral or parasitic infections and experimental immunization with cardiac myosin, the effect of each cytokine on autoimmune myocardial disease may vary. Cytokines may represent new therapeutic targets in the treatment and prevention of autoimmunity-mediated myocarditis and cardiomyopathy, though the etiology and variability in the type of autoimmune responses should be taken into account in the development of cytokine/anti cytokine treatment of these disorders. PMID- 11908774 TI - The transition from viral to autoimmune myocarditis. AB - Myocarditis offers a unique opportunity to study the factors contributing to its transition from a viral infection to an autoimmune disease. In this article, we review recent studies on the role of nitric oxide (NO), gamma interferon (IFN gamma) and interleukin 12 (IL-12) in the progression from early (viral) to late (autoimmune) phases of myocarditis induced by Coxsackievirus B3 (CB3) in highly susceptible (A.CA) and moderately susceptible (B10.M) mice. NO plays a paradoxical role, being protective in early stages but detrimental later in the course of disease. Treatment with antibody to IFN-gamma reduced early disease, but had little effect on the severity of cardiac lesions at later times. Treatment with recombinant (r) IL-12 significantly reduced the autoimmune cardiac lesions in moderately susceptible B10.M mice, but had no measurable effect in highly susceptible A.CA animals. These studies provide evidence that the profile of inflammatory mediators produced early in the course of viral infection determines the later development of autoimmune disease. PMID- 11908775 TI - A peptide fragment of beta cardiac myosin heavy chain (beta-CMHC) can provoke autoimmune myocarditis as well as the corresponding alpha cardiac myosin heavy chain (alpha-CMHC) fragment. AB - The validity of the general belief that alpha cardiac myosin heavy chain (alpha CMHC) is primarily responsible for causing experimental autoimmune myocarditis because of the more profound tolerance induction to beta-CMHC due to its expression during the embryonic stage has been examined. In order to completely avoid cross-contamination among components of the two myosin heavy chains, recombinant myosin fragments were synthesized in Escherichia coli using cDNA fragments of rat alpha- and beta-CMHC cloned by reverse transcription polymerase chain reaction (RT-PCR). Two fragments corresponding to amino acid residues 1107 1164 derived from alpha- and beta-heavy chains were equally capable of provoking severe myocarditis in Lewis rats when immunized in complete Freund's adjuvant. No significant differences in the severity, as judged from histological scoring, were observed between the diseases induced by the two different peptide fragments, indicating conclusively that beta-CMHC is as pathogenic as alpha-CMHC. PMID- 11908776 TI - Heart-infiltrating and peripheral T cells in the pathogenesis of human Chagas' disease cardiomyopathy. AB - Heart tissue destruction in chronic Chagas' disease cardiomyopathy (CCC), occurring in 30% of individuals chronically infected by the protozoan parasite Trypanosoma cruzi, may be caused by autoimmune recognition of patients' heart tissue by a T cell rich inflammatory infiltrate. Recently, our group demonstrated that T cells infiltrating the heart of CCC patients crossreactively recognize cardiac myosin heavy chain and tandemly repetitive T. cruzi antigen B13, and possess an inflammatory T1-type cytokine profile. Susceptibility factors leading 30% of infected patients to develop CCC, while the rest of the patients remain largely asymptomatic (ASY), are still obscure. We compared immunological phenotypes of CCC and ASY patients, who have distinct clinical outcomes despite bearing a similar chronic T. cruzi infection. Preliminary observations indicate that PBMC from CCC patients recognize a set of B13 and cardiac myosin epitopes distinct from that recognized by ASY patients. Moreover, the IFN-gamma response of CCC patients is more intense than that of ASY, both at qualitative and quantitative levels. Taken together, results suggest that heart damage in Chagas' disease cardiomyopathy may be secondary to inflammatory cytokines and a delayed type hypersensitivity process started and/or maintained by heart-crossreactive T cells. Furthermore, the distinct recognition repertoire and the high frequency of IFN-gamma producing among CCC patients could be important factors leading to the differential development of CCC among T. cruzi infected individuals. PMID- 11908778 TI - Cardiac autoantibodies to myosin and other heart-specific autoantigens in myocarditis and dilated cardiomyopathy. AB - Dilated cardiomyopathy (DCM) is characterized by dilation and impaired contraction of the left ventricle or both; it is a relevant cause of heart failure and a common indication for heart transplantation. It may be idiopathic, familial/genetic, viral, autoimmune or immune-mediated, associated with a viral infection. Myocarditis is an inflammatory disease of the myocardium; it may be idiopathic, infectious or autoimmune and may heal or lead to DCM. Thus, in a patient subset, myocarditis and DCM are thought to represent the acute and chronic stages of an organ-specific autoimmune disease of the myocardium. In keeping with this hypothesis, autoimmune features in patients with myocarditis/DCM include: familial aggregation, a weak association with HLA-DR4, abnormal expression of HLA class II on cardiac endothelium on endomyocardial biopsy, detection of organ- and disease-specific cardiac autoantibodies in the sera of affected patients and of symptom-free relatives. The organ-specific cardiac autoantibodies detected by immunofluorescence are directed against multiple antigens. One of these, first identified using immunoblotting and confirmed by ELISA, is the cardiac-specific alpha-myosin isoform. Myosin fulfils the expected criteria for organ-specific autoimmunity, in that immunization with cardiac but not skeletal myosin reproduces, in susceptible mouse strains, the human disease phenotype of myocarditis/DCM; in addition, alpha-myosin is entirely cardiac-specific. The organ-specific cardiac autoantibodies detected by immunofluorescence in symptom-free relatives were associated with echocardiographic features suggestive of early disease. Short-term follow-up is in keeping with this interpretation, although extended follow-up is necessary to define better the role of the antibody as predictor of disease susceptibility in healthy subjects at risk of myocarditis/DCM, such as first-degree relatives. PMID- 11908777 TI - Streptococcus-induced myocarditis in mice. PMID- 11908779 TI - Autoantibodies in Chagas' cardiomyopathy and arrhythmias. PMID- 11908780 TI - Improvement of cardiac function after immunoadsorption in patients with dilated cardiomyopathy. AB - Dilated cardiomyopathy (DCM) is a myocardial disease characterized by progressive depression of myocardial contractile function and by ventricular dilatation. Abnormalities of the cellular and humoral immune system are present in patients with myocarditis and DCM. Various circulating cardiac autoantibodies have been detected among patients suffering from DCM. The relative contribution of cardiac antibodies to cardiac malfunction in DCM remains to be elucidated. Extraction of antibodies by immunoadsorption has been successfully used for treatment of various autoimmune diseases. In this review we report recent studies, which indicate that immunoadsorption improves cardiac function of patients with DCM. The data from these studies indicate that activation of the humoral immune system, with production of cardiac autoantibodies, may play a functional role in cardiac malfunction of patients with DCM. PMID- 11908781 TI - Prevention of experimental autoimmune cardiomyopathy in rabbits by receptor blockers. AB - We investigated the effects of beta1-adrenoceptor blockade and M2-muscarinic receptor antagonist in rabbits which have developed dilated cardiomyopathy-like changes after immunization with the peptides from the second extracellular loop of human beta1-adrenoceptor (beta1-peptide) and M2-muscarinic receptor (M2 peptide). Ten rabbits, which were immunized with beta1-peptide once a month for one year, were treated with bisoprolol and 10 rabbits, which were immunized with M2-peptide, were treated with otenzepad. Although both groups treated with receptor blockade or antagonist showed an increased titer of anti-beta1 adrenoceptor or anti-M2-muscarinic receptor antibodies, myocardial damages were markedly less than those in beta1-peptide- or M2-peptide-immunized rabbits. This study indicates that anti-beta1-adrenoceptor and anti-M2-muscarinic receptor antibodies are of pathogenic importance in the development of human dilated cardiomyopathy, and that beta-adrenoceptor blockade, bisoprolol, and M2 muscarinic receptor antagonist, otenzepad, might be clinically useful for treatment of dilated cardiomyopathy. PMID- 11908782 TI - Structural forces reflecting polyelectrolyte organization from bulk solutions and within surface complexes. AB - The interactions between two macroscopic surfaces approaching one another underlies many of the phenomena observed in Colloid and Interface science. In Russia this gave rise to the branch of colloid science now referred to as Surface Forces. Important discoveries, such as the molecular organization of solvent molecules at an interface, have been unveiled by surface force measurements. More recently, forces and structures at macromolecular length scales have been uncovered. In particular, oscillatory force profiles have been detected from aqueous solutions containing polyelectrolytes. The force-structure relationship can reflect organization in the bulk solution or the internal structure of the adsorbed layer. Using a range of surface force techniques, combined with X-ray and neutron scattering results, we review the main features of these fascinating systems and provide an overview of how they relate to other systems such as micellar solutions, polymer-surfactant complexes and simple solvents. PMID- 11908783 TI - Stability of draining plane-parallel films containing surfactants. AB - The stability of partially mobile draining thin liquid films with respect to axisymmetric fluctuations was studied. The material properties of the interfaces (Gibbs elasticity, surface and bulk diffusions) were taken into account. When studying the long wave stability of films, the coupling between the drainage and perturbation flows was considered and the lubrication approximation was applied. Two types of wave modes were examined: radially-bounded and unbounded waves. The difference between the thickness of loss of stability, h(st), the transitional thickness, h(tr), at which the critical wave causing rupture becomes unstable, and the critical thickness, h(cr), when the film ruptures, is demonstrated. Both the linear and the non-linear theories give h(st) > h(tr) > h(cr). The numerical results show that the interfacial mobility does not significantly influence the thickness of the draining film rupture. The interfacial tension and the disjoining pressure are the major factors controlling the critical thickness. The available experimental data for critical thicknesses of foam and emulsion films show excellent agreement with the theoretical predictions. The important role of the electromagnetic retardation term in the van der Waals interaction is demonstrated. Other published theories of the film stability are discussed. PMID- 11908784 TI - Two-particle dynamics on an electrode in ac electric fields. AB - The relative motion between pairs of negatively charged latex particles 9.7 microm in diameter and deposited on an electrode was measured by optical microscopy and image analysis. At an rms field of approximately 30 V cm(-1), the two particles moved toward each other at frequencies below 500 Hz, but they separated at 1000 Hz. In the cases of aggregation, there are several interesting characteristics. First, when the center-to-center separation of a pair was initially 6 particle radii or more apart, a transient 'incubation' period of tens of seconds was observed before the particles began to move toward each other. Second, the two particles never came into contact, rather at long times the pair maintained a stationary gap between them equal to approximately one-half the particle radius. This stationary gap between particles was also observed for the aggregation of clusters of three or more particles. Finally, the rate of approach for a pair of particles decreased as the frequency increased. Larger fields are required to move particles together in ac compared to dc fields; at 30 Hz the ac field must be 130 times greater than the dc field to achieve the same rate of approach. Taking advantage of the qualitative and quantitative differences of the cooperative motion of particles in dc vs. ac fields, one should be able to re position particles by alternating between these two modes. We demonstrated that the same pair of particles can be brought together at low frequency (100 or 200 Hz) and then separated at high frequency (1000 Hz). PMID- 11908785 TI - Capillary condensation as a morphological transition. AB - The process of capillary condensation/evaporation in cylindrical pores is considered within the idea of symmetry breaking. Capillary condensation/evaporation is treated as a morphological transition between the wetting film configurations of different symmetry. We considered two models: (i) the classical Laplace theory of capillarity and (ii) the Derjaguin model which takes into account the surface forces expressed in terms of the disjoining pressure. Following the idea of Everett and Haynes, the problem of condensation/evaporation is considered as a transition from bumps/undulations to lenses. Using the method of phase portraits, we discuss the mathematical mechanisms of this transition hidden in the Laplace and Derjaguin equations. Analyzing the energetic barriers of the bump and lens formation, it is shown that the bump formation is a prerogative of capillary condensation: for the vapor liquid transition in a pore, the bump plays the same role as the spherical nucleus in a bulk fluid. We show also that the Derjaguin model admits a variety of interfacial configurations responsible for film patterning at specific conditions. PMID- 11908786 TI - Thin wetting films from aqueous electrolyte solutions on SiC/Si wafer. AB - The stability and rupture of thin wetting films from aqueous NaCl or Na2SO4 solutions of different concentrations on silicon carbide were investigated. The flat surface of SiC was obtained by plasma-enhanced chemical vapor deposition (PE CVD) on top of a silicon wafer. The microinterferometric method was used for measuring the film thickness with time. The light reflectance was calculated as a function of film thickness for the four-layer system: air/aqueous solution/SiC/Si wafer. The microinterferometric experiments showed that films from aqueous NaCl and Na2SO4 solutions with concentrations up to 0.01 M were stable independent of the pre-treatment of the substrate. The pre-treatment of the SiC surface was crucial for the wetting film stability at electrolyte concentrations greater than 0.01 M. The films were unstable and ruptured if SiC was washed with 5% hydrofluoric acid and concentrated sulfuric acid, while they were stable if washing was in sulfuric acid only, without immersing SiC in HF. The average equilibrium film thickness was determined as a function of electrolyte concentration. Measurements of the electrokinetic potential zeta were performed by electrophores of SiC powder in 0.001 M NaCl. It was shown that silicon carbide surface was negatively charged. The theory of heterocoagulation was used for the interpretation of the results. Besides the DLVO forces, the structural disjoining pressure (both positive and negative) has been included in the analysis. PMID- 11908787 TI - The influence of solid-liquid interactions on dynamic wetting. AB - The molecular-kinetic theory of dynamic wetting has been extended to take specific account of solid-liquid interactions. By equating the work of adhesion with the surface component of the specific activation free energy of wetting, equations have been derived which show the way in which solid-liquid interactions modify both the driving force and the resistance to wetting. For a liquid meniscus advancing across the surface of a solid, these two effects have opposing consequences. Thus, strong interactions increase both the driving force and the resistance, while weak interactions decrease the driving force and the resistance. Because of the form of the relationships, the two effects do not simply cancel out. As a result, the maximum rate at which a liquid can wet a solid may exhibit its own maximum at some intermediate level of interaction. Data taken from both experimental and molecular-dynamics simulations are shown to support these findings, which have significant implications for any process where wetting dynamics are important, such as coating. PMID- 11908788 TI - Capillary forces between surfaces with nanoscale roughness. AB - The flow and adhesion behavior of fine powders (approx. less than 10 microm) is significantly affected by the magnitude of attractive interparticle forces. Hence, the relative humidity and magnitude of capillary forces are critical parameters in the processing of these materials. In this investigation, approximate theoretical formulae are developed to predict the magnitude and onset of capillary adhesion between a smooth adhering particle and a surface with roughness on the nanometer scale. Experimental adhesion values between a variety of surfaces are measured via atomic force microscopy and are found to validate theoretical predictions. PMID- 11908789 TI - Disjoining pressures, zeta potentials and surface tensions of aqueous non-ionic surfactant/electrolyte solutions: theory and comparison to experiment. AB - A self-consistent electrostatic theory is presented to predict disjoining pressure isotherms of aqueous thin-liquid films stabilized by non-ionic surfactants and air/water surface tensions and zeta potentials of electrolyte solutions with and without non-ionic surfactant. The proposed model combines specific adsorption of hydroxide ions at the interface with image charge and dispersion forces on ions in the diffuse double layer. The result is a quantitative description of aqueous solution interfaces as a function of surfactant concentration, ionic strength and pH. Disjoining pressure isotherms of thin-liquid films stabilized by non-ionic surfactants and electrophoresis experiments on air bubbles and oil droplets in aqueous solutions demonstrate that hydroxide ions specifically adsorb at air/water and oil/water interfaces. The surface charge increases with pH, decreases with increasing surfactant concentration, increases slightly with ionic strength, and for n-alkyl polyethylene oxide non-ionic surfactants is not significantly affected by surfactant molecular structure. Concentrated electrolyte-solution surface tensions, however, indicate that ions are repelled from the air/water interface by an 'image charge' force, that is a parameterized by the ion valence and the ionic strength of the aqueous solution. Additionally, differences in induced induced dipole forces on an ion near an interface lead to a van der Waals dispersion interaction force that depends on the ion polarizabilites and the molecular properties of the two surrounding bulk phases. Incorporation of these two additional ion free energies into the Poisson-Boltzmann equation along with a simple model for hydroxide-ion specific adsorption at the air/water interface results in a non-linear second-order ordinary differential equation containing two adjustable parameters. The proposed modified Poisson-Boltzmann (MPB) theory accurately predicts newly measured disjoining pressures of thin-liquid foam films stabilized by polyethylene oxide n-alkyl ether surfactants. With no additional adjustable parameters, zeta potentials of nascent air bubbles in water and surface tensions of aqueous electrolyte solutions are successfully predicted. The new electrostatic model also explains the fascinating existence of a surface tension minimum in dilute electrolyte solutions, known as the Jones-Ray effect. PMID- 11908790 TI - Electrokinetic properties of methylated quartz capillaries. AB - Electrokinetic (zeta)-potentials of methylated (trimethylchlorosilane) quartz capillaries (5-6 microm in radius) were determined in 10(-4) M KCl solution. Over the course of time, the absolute values of the zeta-potential decrease, as a result of the formation of small bubbles on the rough methylated surface, generated from the flowing, nitrogen gas-saturated solution. This decrease is attributed to screening of a part of the solid surface. After the passage of time, a sharp increase in the zeta-potentials was observed, as the pressure was increased and the initial potential values were recovered. Sometimes, oscillations in the zeta-potentials were observed. This behaviour was explained by detachment of bubbles from the methylated surfaces by the flowing solution. Addition of non-ionic surfactant, which made the methylated surface hydrophilic, decreased the measured zeta-potentials. This was attributed to suppression of water slippage, an effect known to occur for hydrophobic solid surfaces. A mixed mechanism of charge formation is characteristic for these methylated quartz surfaces and is connected with presence of hydrophobic and hydrophilic areas. The ratio between these areas controls both the formation of surface charge as well as the contact angles. PMID- 11908792 TI - Adsorption of single and mixed ionic surfactants at fluid interfaces. AB - Two different approaches have been used in the literature to describe the effects of ionisation of surfactants on the surface pressure, pi. One approach is based on a molecular model for a charged monolayer, in which the mutual repulsion of the long-chain surfactant ions results in an additional surface pressure, piel, calculated with the Gouy-Chapman theory for the formation of a diffuse electric double layer, and with counterion binding in the Stern-Helmholtz layer adjacent to the surfactant monolayer. The other approach regards the surface as a two dimensional solution defined as a Gibbs dividing surface, which is electroneutral by definition. In this approach, the adsorption of any ion is the sum of its excesses in the monolayer and the electrical double layer; no assumptions are made about the spatial distribution of charges. It has been shown that both models can produce a reasonable description of experimental results obtained for solutions of a single ionic surfactant (RX) with or without inorganic electrolyte (XY). In many cases, measurements of pi vs. mean ionic activity at different salt concentrations (cXY) are found to coincide on a single curve, implying that at given mean ionic activity both adsorption and pi are independent of cXY, i.e. that double-layer contributions to the surface pressure are negligible. In addition, the electroneutral 2-D solution approach has resulted in a simple explanation of several typical features of mixed ionic surfactant solutions, in particular for mixtures of anionic and cationic surfactants. In mixed solutions too, double-layer effects appear to be negligible. We present arguments for such negligibility. One reason is a significant degree of binding between adsorbed surface active ions (R) and counterions (X); another is that for 1:1 electrolytes, the contribution of the diffuse double layer to the adsorption of the combination (RX) vanishes. As a result, it is possible to interpret the same experimental data in terms of both models. PMID- 11908791 TI - Viscosity of concentrated suspensions: influence of cluster formation. AB - Dispersed particles can form clusters even at low concentrations. Colloidal and hydrodynamic forces are responsible for this phenomenon and these forces determine both structure and size of clusters. We assume that the viscosity of a concentrated suspension is completely determined by cluster size distribution, regardless if clusters form under the action of colloidal, hydrodynamic interactions or applied shear rates. Based on this assumption an equation, which describes dependency of viscosity on a concentration of dispersed particles taking into account cluster formation, is deduced. Under special restrictions the deduced dependency coincides with the well-known Dougherty-Krieger's equation except for a clear physical meaning of parameters entered. Our consideration shows that Dougherty-Krieger's equation has deeper physical background than it has been supposed earlier. Experimental verification of the suggested model shows a good agreement with the theory predictions and proves a presence of clusters even at low concentrations of dispersed particles. PMID- 11908793 TI - Superspreading driven by Marangoni flow. AB - The spontaneous spreading (called superspreading) of aqueous trisiloxane ethoxylate surfactant solutions on hydrophobic solid surfaces is a fascinating phenomenon with several practical applications. For example, the ability of trisiloxane ethoxylate surfactants to enhance the spreading of spray solutions on waxy weed leaf surfaces, such as velvetleaf (Abutilion theophrasti), makes them excellent wetting agents for herbicide applications. The superspreading ability of silicone surfactants has been known for decades, but its mechanism is still not well understood. In this paper, we suggest that the spreading of trisiloxane ethoxylates is controlled by a surface tension gradient, which forms when a drop of surfactant solution is placed on a solid surface. The proposed model suggests that, as the spreading front stretches, the surface tension increases (the surfactant concentration becomes lower) at the front relative to the top of the droplet, thereby establishing a dynamic surface tension gradient. The driving force for spreading is due to the Marangoni effect, and our experiments showed that the higher the gradient, the faster the spreading. A simple model describing the phenomenon of superspreading is presented. We also suggest that the superspreading behavior of trisiloxane ethoxylates is a consequence of the molecular configuration at the air/water surface (i.e. small and compact hydrophobic part), as shown by molecular dynamics modeling. We also found that the aggregates and vesicles formed in trisiloxane solutions do not initiate the spreading process and therefore these structures are not a requirement for the superspreading process. PMID- 11908795 TI - Forces due to dynamic structure in thin liquid films. AB - The long-range surface forces arising due to dynamic structure in thin liquid films are considered. The phonon formalism is applied to analyse the contribution of the collective motions in liquid to the excess free energy and the disjoining pressure of the film. It is shown that both the intermolecular interactions and the interactions between liquid and confining phases essentially have an influence on the sign and the magnitude of the phonon component of disjoining pressure and its temperature dependence. The methods and procedures used to assess the density of vibrational states for real and model liquids are discussed. The description of several spectroscopic experimental methodics specific for investigation of thin liquid interlayers is given concurrently with experimental data revealing the dynamic structuring in the course of film thinning. PMID- 11908794 TI - Simple model for prediction of surface tension of mixed surfactant solutions. AB - A rigorous theoretical model is presented which describes the equilibrium behaviour of a surfactant mixtures at liquid/fluid interfaces. The theory describes mixtures of surfactants with different molar areas and accounts for the non-ideality of the surface layer. The theoretical results are in good agreement with experimental data and support the idea of additivity of the interaction parameters in the surface layer. The rigorous equation of state is transformed into simple relationships for the description of the adsorption behaviour of mixed surfactant systems. The model requires surface tensions of the single surfactant systems or the adsorption isotherms to construct the isotherm of the mixture while no extra interaction parameters between the different compounds are assumed. The model is tested with a number of literature data, such as mixed sodium alkyl sulfates, mixtures of betaine homologues BHB12 with BHB16, non-ionic surfactant mixtures, and anionic-nonionic mixtures (1-butanol with BHB12, and oxethylated decanol (C10EO5) with sodium dodecyl sulfate). The agreement between experimental data and the theoretical calculations is excellent. This approach can be especially important for practical applications of surfactant mixtures for which experimental data are scarce. PMID- 11908796 TI - Films driven by surface tension gradients. AB - Fingering instabilities are often observed at the contact line of drops of surfactant solutions spreading spontaneously on solid surfaces. It has been recognised recently that a usual linear stability analysis predicts stable behaviour in contrast to the observed instability. It now seems the instability arises from short-time transients, where the thickness of the film ahead of the main drop is a crucial parameter for amplification. We reconsidered previous experiments and performed new ones along these lines. The strengths and weaknesses of the available models were analysed. PMID- 11908797 TI - Chain-melting phase transition and short-range molecular interactions in phospholipid foam bilayers. AB - Occurrence of two-dimensional chain melting phase transition in foam bilayers was established for the first time. Microscopic horizontal foam bilayers [Newton black films (NBF)] were investigated by the microinterferometric method of Scheludko-Exerowa. The foam bilayers were formed from water-ethanol solutions of dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) and egg phosphatidylcholine (Egg PC) and samples of amniotic fluid (AF) at different temperatures. The influence of temperature on the foam bilayer thickness h(w) and on the critical concentration Cc for formation of foam bilayer was studied. It was shown that in the range of the main phase transition the temperature dependence of h(w) and C(c) changed specifically in the case of DMPC and DPPC foam bilayers. The thickness of the foam bilayers increased with decreasing temperature in the range of the main phase transition due to the melting of hydrocarbon tails of phospholipid molecules. These changes took place at the temperatures of the bulk chain-melting phase transitions, as determined by differential scanning calorimetry (DSC) for both aqueous, and water/ethanol DMPC, DPPC, and DPPC dispersions. An effect of the 'disperse medium' on h(w) was found for foam bilayers from DPPC. The results that foam bilayers could have different thickness at different temperatures disproved the current concept that NBF acquired constant thickness at concentrations higher than C(el,cr). The data for Cc were analysed on the basis of the hole-nucleation theory of bilayer stability of Kashchiev and Exerowa. This theory considered the amphiphile bilayer as a two dimensional ordered system with short-range molecular interactions between the first neighbour molecules (as in a crystal). The short-range molecular interactions were presented by the parameter binding energy Q of an amphiphile molecule in the bilayer. The binding energy Q of two neighbouring phospholipids was calculated for the gel (30-60 kT) and liquid crystalline state (16-18 kT) of the bilayers from DMPC, DPPC, Egg PC, AF. Concentration/temperature phase diagram of DPPC foam bilayers that defined regions of gaseous (ruptured), gel and liquid crystalline foam bilayers were drawn. The values of Q obtained for various samples were very close and vary from 5.3 x 10(-20) to 9.4 x 10(-20) (approx. 13 22 kT) which indicated that in all cases the foam bilayers were in liquid crystalline state. This is an important result since the parameter studied threshold concentration (threshold dilution) is crucial for a very successful assessment of the risk for respiratory distress syndrome (RDS) in newborns and could be employed in medicine for assessment of other respiratory disturbances. It is to be expected that foam bilayers from phospholipids could be used as a model for investigation of short-range forces in biological structures, of interaction between membranes, etc. PMID- 11908798 TI - Charge effects for differentiation of oligodeoxynucleotide isomers containing 8 oxo-dG residues. AB - Dissociation reactions of a series of multiply charged oligodeoxynucleotide (ODN) 12-mer anions were studied using an ion trap mass spectrometer. These mixed nucleobase 12-mers fragment first by loss of a neutral nucleobase (A, G, C, and/or 5-methyl-cytosine) followed by cleavage at 3' C-O bond of the sugar from which the base is lost to produce the complementary sequence ions, i.e., [a - B] and w type of ions. No detectable loss of 8-oxo-guanine and/or thymine from these 12-mers is observed under gentle collision conditions in the ion trap. The primary loss of a nucleobase and the subsequent backbone cleavage to generate sequence ions strongly depend on the charge state of the parent molecular ion. For low charge states (2- and 3-), product ions due to the loss of a neutral guanine base and related sequence ions are dominant in the tandem mass spectra. However, preferential loss of a neutral adenine becomes the primary reaction channel from the 5- charge state of the molecular ion. Such charge state dependent fragmentation behavior was utilized to determine the site of 8-oxo-dG residue in a series of structural isomers. The position of 8-oxo-dG residue can be simply determined from the fragmentation pattern of 3- charge state, but not of 5- charge state. It is suggested that in addition to specific modification that affects the N-glycosidic bond strength, total charge content of an ODN is an important factor for determining the differential fragmentation behavior. PMID- 11908799 TI - SOS: a simple interactive program for ab initio oligonucleotide sequencing by mass spectrometry. AB - Mass spectra of oligonucleotides derived from collision-induced dissociation following electrospray ionization provide an effective means of sequence determination, at the 20-mer level and below. An interactive, stand-alone computer program, Simple Oligonucleotide Sequencer (SOS) has been developed for rapid oligonucleotide sequencing from mass spectra, under user control on a residue by residue basis. Modifications can be defined in any combination for the base, sugar or backbone. Sequence ladders can be independently constructed in both the 5' --> 3' directions and 3' --> 5' directions, and graphically compared for homology and overlap. A particular advantage of this method is the ability to easily erase and rebuild alternate subsequences. The program can be used for ab initio sequencing of modified or unmodified oligonucleotides, for rapid verification of sequence, and in studies of fragmentation processes of model oligonucleotide derivatives. PMID- 11908800 TI - Comparison of SSI with APCI as an interface of HPLC-mass spectrometry for analysis of a drug and its metabolites. AB - Sonic spray ionization (SSI) was compared with atmospheric pressure chemical ionization (APCI) as an interface of high-performance liquid chromatography (HPLC)-mass spectrometry (MS) for sensitive analyses of a neuroleptic drug, haloperidol and its two metabolites, such as reduced haloperidol and 4-(4 chlorophenyl)-4-hydroxypiperidine (CPHP), in biological samples. For both SSI and APCI interfaces, HPLC-MS-MS gave higher sensitivity than HPLC-MS. The sensitivities by HPLC-SSI-MS-MS for haloperidol and reduced haloperidol were 100 and 30 times higher, respectively, than those by HPLC-APCI-MS-MS; no spectrum with recognizable peaks was obtained for CPHP with the APCI interface. Therefore, detection limits and regression equations were examined by the HPLC-SSI-MS-MS for human plasma and urine samples spiked with the above drug and its metabolites. Haloperidol, reduced haloperidol, and CPHP showed good linearity in the ranges of 5-800, 10-800, and 100-800 ng/mL, respectively, for both human plasma and urine; their detection limits were 2.5, 5, and 75 ng/mL, respectively, using a new polymer HPLC column which enabled direct application of biological samples. PMID- 11908801 TI - Measurements of mean initial velocities of analyte and matrix ions in infrared matrix-assisted laser desorption ionization mass spectrometry. AB - The mean initial velocities of analyte ions ranging in molecular weight from 1000 Da to 150 kDa and desorbed with a pulsed Er:YAG laser from various solid-state and liquid IR MALDI matrices were measured along with those of the matrix ions. Experiments with UV MALDI were performed for comparison in addition for a 2,5 dihydroxybenzoic acid preparation. Two different measurement principles were employed, (1) a delayed extraction method, relying on the initial velocity dependent increase of flight times with delay time between laser and HV ion extraction pulse, and (2) a field-free drift method in which the first region of a two-stage ion source was varied in length and the flight times compared. The two methods yielded somewhat different values for the mean initial ion velocities. Based on a detailed discussion of the measurement principles it is suggested that the actual initial velocities of IR MALDI ions lie between the limits set by the two methods. The influences of the analyte-to-matrix ratio, laser fluence, and laser wavelength on the initial ion velocities were also investigated. Significant differences between the desorption mechanisms for liquid and solid-state matrices were observed. PMID- 11908802 TI - Visualization and analysis of molecular scanner peptide mass spectra. AB - The molecular scanner combines protein separation using gel electrophoresis with peptide mass fingerprinting (PMF) techniques to identify proteins in a highly automated manner. Proteins separated in a 2-dimensional polyacrylamide gel (2-D PAGE) are digested in parallel and transferred onto a membrane keeping their relative positions. The membrane is then sprayed with a matrix and inserted into a matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometer, which measures a peptide mass fingerprint at each site on the scanned grid. First, visualization of PMF data allows surveying all fingerprints at once and provides very useful information on the presence of chemical noise. Chemical noise is shown to be a potential source for erroneous identifications and is therefore purged from the mass fingerprints. Then, the correlation between neighboring spectra is used to recalibrate the peptide masses. Finally, a method that clusters peptide masses according to the similarity of the spatial distributions of their signal intensities is presented. This method allows discarding many of the false positives that usually go along with PMF identifications and allows identifying many weakly expressed proteins present in the gel. PMID- 11908804 TI - Characterization of amino acid side chain losses in electron capture dissociation. AB - We have used electrospray ionization (ESI) Fourier-transform ion cyclotron resonance (FTICR) mass spectrometry to characterize amino acid side chain losses observed during electron capture dissociation (ECD) of ten 7- to 14-mer peptides. Side-chain cleavages were observed for arginine, histidine, asparagine or glutamine, methionine, and lysine residues. All peptides containing an arginine, histidine, asparagine or glutamine showed the losses associated with that residue. Methionine side-chain loss was observed for doubly-protonated bombesin. Lysine side-chain loss was observed for triply-protonated dynorphin A fragment 1 13 but not for the doubly-protonated ion. The proximity of arginine to a methoxy C-terminal group significantly enhances the extent of side-chain fragmentation. Fragment ions associated with side-chain losses were comparable in abundance to those resulting from backbone cleavage in all cases. In the ECD spectrum of one peptide, the major product was due to fragmentation within an arginine side chain. Our results suggest that cleavages within side chains should be taken into account in analysis of ECD mass spectral data. Losses from arginine, histidine, and asparigine/glutamine can be used to ascertain their presence, as in the analysis of unknown peptides, particularly those with non-linear structures. PMID- 11908805 TI - Generation and characterization of ionic and neutral P(OH)2+/* in the gas phase by tandem mass spectrometry and computational chemistry. AB - The bicoordinated dihydroxyphosphenium ion P(OH)2+ (1+) was generated specifically by charge-exchange dissociative ionization of triethylphosphite and its connectivity was confirmed by collision induced dissociation and neutralization-reionization mass spectra. The major dissociation of 1+ forming PO+ ions at m/z 47 involved another isomer, O=P-OH2+ (2+), for which the optimized geometry showed a long P-OH2 bond. Dissociative 70-eV electron ionization of diethyl phosphite produced mostly 1+ together with a less stable isomer, HP(O)OH+ (3+). Ion 2+ is possibly co-formed with 1+ upon dissociative 70 eV electron ionization of methylphosphonic acid. Neutralization-reionization of 1+ confirmed that P(OH)2* (1) was a stable species. Dissociations of neutral 1, as identified by variable-time measurements, involved rate-determining isomerization to 2 followed by fast loss of water. A competitive loss of H occurs from long-lived excited states of 1 produced by vertical electron transfer. The A and B states undergo rate-determining internal conversion to vibrationally highly excited ground state that loses an H atom via two competing mechanisms. The first of these is the direct cleavage of one of the O-H bonds in 1. The other is an isomerization to 3 followed by cleavage of the P-H bond to form O=P-OH as a stable product. The relative, dissociation, and transition state energies for the ions and neutrals were studied by ab initio and density functional theory calculations up to the QCISD(T)/6-311+G(3df,2p) and CCSD(T)/aug-cc-pVTZ levels of theory. RRKM calculations were performed to investigate unimolecular dissociation kinetics of 1. Excited state geometries and energies were investigated by a combination of configuration interaction singles and time-dependent density functional theory calculations. PMID- 11908803 TI - Solution composition and thermal denaturation for the production of single stranded PCR amplicons: piperidine-induced destabilization of the DNA duplex? AB - Strategies to produce single-stranded PCR amplicons for detection by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) were investigated using modified electrospray solutions and by thermally denaturing the duplex structures with a resistively heated electrospray ionization source. A synthetic 20-mer oligonucleotide annealed to its complementary strand was used as a model system for initial experiments. Electrospray solutions were altered by varying the relative proportion of aqueous phase in efforts to induce destabilization of the double helix. When the electrospray solution contains a 25% aqueous content, the 20-mer oligonucleotide is detected in its double-stranded form. Increasing the proportion of aqueous phase in the electrospray solution to 60% destabilized the double helix, resulting in the detection of only single-stranded species. This strategy was extended to an 82-bp polymerase chain reaction (PCR) product derived from the human tyrosine hydroxylase gene (HUMTH01). In efforts to destabilize the 82-bp PCR product, electrospray solutions reaching 70% aqueous content were necessary to promote the detection of only single-stranded amplicons. Implementation of the resistively heated transfer line and an electrospray solution in which the oligonucleotide is on the threshold of duplex stability allowed for double stranded and single-stranded species to be generated from the same ESI solutions at both ambient and elevated transfer line temperatures, respectively, without disruption of the electrospray process. The volatile base piperidine, present at 20 mM concentrations in the electrospray solution, was found to play a critical role in the formation of single-stranded species at the higher aqueous percentages and a duplex destabilization mechanism has been proposed. PMID- 11908806 TI - A simple approach for coupling liquid chromatography and electron ionization mass spectrometry. AB - A miniaturized, aerosol based interface for directly coupling a liquid chromatograph with a mass spectrometer is presented. The interface is entirely within the electron ionization (EI) source of the mass spectrometer and no additional, external devices are needed. This simple and effective approach exploits micro-flow nebulization technology providing a new interface suitable for a variety of applications of environmental and biological interest. The new interface provides necessary linearity, ruggedness, sensitivity, and reproducibility of response for trace level analysis, and readily interpretable mass spectra for unambiguous identification of unknown compounds of small to medium molecular weight. PMID- 11908807 TI - Fragmentation of phosphopeptides by atmospheric pressure MALDI and ESI/Ion trap mass spectrometry. AB - An investigation of phosphate loss from phosphopeptide ions was conducted, using both atmospheric pressure matrix-assisted laser desorption/ionization (AP MALDI) and electrospray ionization (ESI) coupled to an ion trap mass spectrometer (ITMS). These experiments were carried out on a number of phosphorylated peptides in order to investigate gas phase dephosphorylation patterns associated with phosphoserine, phosphothreonine, and phosphotyrosine residues. In particular, we explored the fragmentation patterns of phosphotyrosine containing peptides, which experience a loss of 98 Da under collision induced dissociation (CID) conditions in the ITMS. The loss of 98 Da is unexpected for phosphotyrosine, given the structure of its side chain. The fragmentation of phosphoserine and phosphothreonine containing peptides was also investigated. While phosphoserine and phosphothreonine residues undergo a loss of 98 Da under CID conditions regardless of peptide amino acid composition, phosphate loss from phosphotyrosine residues seems to be dependent on the presence of arginine or lysine residues in the peptide sequence. PMID- 11908809 TI - C1 inhibitor deficiency: hereditary and acquired forms. AB - C1 inhibitor deficiency can be hereditary or acquired. The hereditary disorder has two types, each of which is inherited as a dominant disorder, with genetic mechanisms leading either to low levels of normal C1 INH and little or no mutant problem as a result of mRNA or protein synthetic defects or degradative mechanisms (Type I) or with point mutations and synthesis of a functionless protein product with transinhibition of the normal allele (Type II). The acquired disorder with low C1q is due to C1 INH consumption associated with lymphoma or connective tissue disease (Type I) and/or autoimmune mechanisms (Type II). The swelling of all types is due to absence of inhibition of the plasma kinin forming cascade with liberation of bradykinin while complement activation, a critical marker of the disorder, is not responsible for the swelling. Treatment employs androgenic compounds, antifibrinolytic agents, or replacement therapy. PMID- 11908808 TI - Conformational studies of Zn-ligand-hexose diastereomers using ion mobility measurements and density functional theory calculations. AB - Ion mobility studies and density functional theory calculations were used to study the structures of [Zn/diethylenetriamine/Hexose/Cl]+ complexes in an effort to probe differences in the three-dimensional conformations. This information allows us to gain insight into the structure of these complexes before collisional activation, which is the first step in understanding the stereoselective dissociations observed under collisionally activated conditions. The collision cross sections obtained from the ion mobility measurements showed that the mannose structure is more compact than the galactose and glucose complexes, respectively. Using density functional theory, candidate structures for each of the experimentally observed complexes were generated. Two criteria were used to determine the most likely structures of these complexes before activation: (1) The allowed relative energies of the molecules (between 0-90 kJ/mol) and (2) collision cross section agreement (within 2%) between the theoretically determined structures and the experimentally determined cross section. It was found that the identity of the monosaccharide made a difference in the overall conformation of the metal-ligand-monosaccharide complex. For glucose and galactose, metal coordination to O(6) was found to be favorable, with the monosaccharide occupying the 4C1 chair conformation, while for mannose, O(2) metal coordination was found with the monosaccharide in a B3,0 conformation. Coordination numbers varied between four and six for the Zn(II) metal centers. Given these results, it appears that the stereochemistry of the monosaccharide influences the conformation and metal coordination sites of the Zn(II)/monosaccharide/dien complex. These differences may influence the dissociation products observed under collisionally activated conditions. PMID- 11908811 TI - Compilation and meta-analysis of randomized placebo-controlled clinical trials on the prevention of respiratory tract infections in children using immunostimulants. AB - BACKGROUND: Several immunostimulants presume to prevent respiratory tract infections (RTIs) in children, but their efficacy is controversial. OBJECTIVES: To compile the findings of the randomized, placebo-controlled trials (RCTs) on the prevention of acute respiratory tract infections (ARTIs) in children using immunostimulants, and to perform a meta-analysis. DATA SOURCES: Medline, EMBASE databases, and register of Cochrane Acute Respiratory Infection Group. REVIEW METHODS: We searched all the references of immunostimulants and selected papers referring to RCTs on the prevention of ARTIs in children. Papers were rated according to Jadad's instrument. We abstracted the number of ARTIs, and a one tailed probability value (p) was abstracted for each trial. Effect of medication was determined as weighted mean +/- SE of percent reduction of ARTIs regarding ARTIs of placebo groups as 100%. RESULTS: Four of five RCTs with Jadad's score > 3 showed significant reduction of ARTIs in immunostimulant groups. When only the trials reporting mean +/- SD and/or dispersion were considered (n = 16), the global weighted percent effect of immunostimulants showed a change of -42.64%, with 95% confidence intervals from -45.19% to -40.08%; i. e., the treated group presented about 60% of the mean number of ARTIs in the placebo group. CONCLUSIONS: According to this meta-analysis and RCTs with Jadad's score > 3, immunostimulants are an effective treatment for the prevention of ARTI. Further high-quality RCTs are required to demonstrate the effect and the size of the effect of each individual immunostimulant. PMID- 11908810 TI - Pulmonary remodeling in asthma. AB - For many years it was assumed that all asthmatics had an at least potentially reversible disease. It is now clear both from longitudinal studies of FEV1 and biopsy data that some asthmatics develop permanent obstructive lung disease. Some adults exhibit an accelerated decline in lung function, and some children never reach normal lung volume. The most likely histologic changes accounting for this phenomena are the deposition of collagen and glycoprotein beneath the basement membrane and in the extracellular matrix, and the destruction of elastic tissue. This permanent obstruction does not occur in all asthmatics. Factors that place patients at increased risk appear to be related to the severity and the duration of the disease and the degree of bronchial hyperresponsiveness. It is unclear, based upon present data, as to whether or not inhaled corticosteroids can favorably affect the process of remodeling, but evidence seems to favor this hypothesis. Inhaled corticosteroids, in some studies, have been shown to decrease the thickness of the lamina reticularis and retard the decline in FEV1. In addition, removal of the source of asthma, as demonstrated in occupational asthma due to toluene diisocyanate, can have a beneficial effect in this regard. PMID- 11908812 TI - Study of the in vitro sulphidoleukotriene production in food-allergic patients. AB - OBJECTIVE AND DESIGN: To study in vitro sulphidoleukotriene (sLT) production by food allergic patients using cellular allergen stimulation test (CAST)-ELISA and to evaluate the reliability of this technique for diagnosing food allergic reactions. SUBJECTS: Forty patients with adverse reactions after food intake, 20 healthy controls, and 15 individuals sensitized to inhalant allergens as atopic controls. METHODS: Skin tests, serum-specific IgE, histamine release test (HRT), CAST-ELISA and food challenges. One-way ANOVA was used to compare tests results between patients and controls and to study mediator release and specific IgE, related to the severity of clinical pictures. Sensitivity and specificity were analyzed by ROC curves. RESULTS: Food allergic patients showed higher (p < 0.05) Ag-dependent sLT production (836.2 +/- 664.1 pg/ml) (mean +/- standard deviation) than both control groups. After stimulus with anti-IgE antibodies, sLT production was higher (p < 0.05) by atopic controls (1630.8 +/- 696.5 pg/ml) compared to patients and healthy controls. Patients with anaphylactic reactions showed higher Ag-specific and anti-IgE sLT and histamine production than patients with less severe manifestations. Mean serum-specific IgE was significantly lower (p < 0.05) in patients presenting oral allergy syndrome compared to patients with more severe clinical pictures. CAST-ELISA was the most sensitive method. Prick by prick test was the most specific. CONCLUSIONS: CAST-ELISA may provide a useful tool for diagnosing food allergy. Enhanced cell releasability may be linked to the severity of the clinical response to foods. PMID- 11908813 TI - Immunoblot studies in birch pollen-allergic patients with and without fruit hypersensitivity: part I: antibody pattern for birch pollen extract. AB - About 40-70% of birch pollen allergic patients show allergic symptoms after ingesting or handling raw fruits. Several investigations have indicated a partial immunological identity between birch pollen and stone fruit. To further clarify this association, we investigated 59 patients with allergic symptoms (conjunctivitis, rhinitis, and asthma during the birch pollen season) and 18 nonatopic controls by skin prick test (SPT) and RAST with birch pollen, fresh apple, cherry, and peach as well as freshly prepared fruit extracts. According to a questionnaire dealing with symptoms after ingestion of raw fruits, the subjects were divided into groups with (35 FH+) and without (24 FH-) fruit hypersensitivity. IgE, IgG, IgG1, IgG4, IgA, and IgM binding patterns to birch pollen extracts were performed with 33 sera (12 FH+, 11 FH-, and 10 nonatopic controls) using the immunoblot-technique. Patients with FH+ expressed a significantly stronger sensitization to birch pollen than patients without FH-, as measured by RAST and SPT. Native fruits induced stronger SPT reactions than fruit extracts, and patients with FH+ showed a significantly higher skin index with all fruits and fruit extracts tested. Specific IgE, IgG, IgG1, IgG4, IgM and IgA to birch pollen extracts could be detected by immunoblot in all groups, albeit with different frequencies and intensities. From this data we conclude that fruit hypersensitivity is related more to the 17 kd and 67-85 kd than to the 26-28 kd or 36 kd protein bands of the birch pollen extract. The relationship of specific IgE > IgG > IgM to a single protein band seems to be associated with the development of symptomatic type I allergy. PMID- 11908814 TI - Immunoblot studies in birch pollen-allergic patients with and without fruit hypersensitivity: part II: antibody pattern for fruit extracts. AB - Patients allergic to birch pollen also exhibit more hypersensitivity reactions to fresh fruits and vegetables than do patients allergic to other pollens. Several investigations have indicated a possible partial immunological identity between birch pollen and fruits. To study this, 23 birch pollen-allergic patients 12 with (FH+) and 11 without (FH-) fruit hypersensitivity and 10 nonatopic controls were examined with self-prepared apple-peel, cherry, and peach extracts by immunoblotting. The self-prepared extracts were characterized by histamine release studies with 20 FH+ birch pollen-allergic patients. Specific IgE, IgG, IgG1, IgG4, IgA, and IgM binding patterns of the fruit extracts presented an individual distribution with at least 1-3 IgE bands at varying molecular weight locations. The FH+ group expressed intense IgE binding to the different extracts compared to the FH- group, and even the control group showed all immunoglobulin classes, though different frequencies and intensities compared to the allergic groups. It seemed that the specific IgE > IgG > IgM relation to a single antigen is important for distinguishing between symptomatic and asymptomatic persons. With this hypothesis we found most IgE with less IgG binding to apple-peel in the region of 22-28 kd and 43-56 kd, cherry: 15-25 kd and 72- > 90 kd and peach 35-41 kd and 66-76 kd, suggesting that these proteins might be important for cross reactivity with birch pollen and developing fruit hypersensitivity. PMID- 11908815 TI - Prevalence of latex sensitivity in patients with malignancies. AB - An increased frequency of allergic reactions to latex have been reported in specific populations with chronic latex exposure (e. g., children with spina bifida, health-care workers, rubber and doll-manufacturing workers). However, latex sensitization occurs in adult patients with no known risk factors. This study investigated hypersensitivity to latex in patients with malignancies. MATERIAL AND METHODS: We studied 70 adult patients (40 females, 30 males) with a standard questionnaire, skin prick tests (SPTs) with latex, common aeroallergens and fruits to all of the subjects. In addition, specific IgE concentrations for all of these allergens were measured in serum with the Pharmacia UniCAP system. RESULTS: The prevalence of latex sensitization was found to be 11.4% (n = 8). Both SPTs and anti-latex IgE antibodies of these patients were positive in the sera. Latex sensitization was related to the personal history of allergic diseases (p = 0.02), the duration of disease (p = 0.01), and the number of invasive procedures (p = 0.04). DISCUSSION: This study shows that latex hypersensitivity is present in patients with malignancies. From our results latex sensitization must be kept in mind while dealing with this susceptible patient group. PMID- 11908816 TI - T-cell reactions to drugs in distinct clinical manifestations of drug allergy. AB - BACKGROUND: Recent data indicate that T cells play a major role in different forms of drug allergies. OBJECTIVE: To show that T-cell reactions are involved in various forms of adverse reactions to different kinds of drugs, and that lymphocyte transformation and skin tests may be positive in patients who had distinct clinical manifestations of drug allergies. METHODS: We collected data of 44 patients with a highly suggestive history for adverse drug reaction who had on subsequent investigations a positive lymphocyte transforrmation test. In 41/44 patients (93%) skin tests with the suspected drugs were performed and in some cases drug-specific IgE-antibodies were determined. All patients were HLA typed. RESULTS: Clinical manifestations of the drug allergy were heterogeneous, comprising maculopapular and bullous exanthema, erythema exsudativum multiforme, vasculitis, serum sickness, urticaria, as well as involvement of internal organs. Maculopapular exanthemas formed the largest group (54%), followed by reactions more indicative of immediate hypersensitivity (28%), such as urticaria/angioedema. In most cases (63%), beta-lactam antibiotics were found to have caused the allergic reaction. Skin tests for immediate reactions were positive in 6/40 patients (15%) tested, those for late reactions in 24/38 patients (63%) tested. CONCLUSIONS: Our data provide evidence that drug-specific T cells can be detected in distinct clinical manifestations of drug allergy. A combined approach using a detailed case history, lymphocyte transformation tests, skin tests (immediate and delayed type) appears to be helpful to identifying the incriminated drug. PMID- 11908818 TI - The influence of a central vacuum system on quality life in patients with house dust-associated allergic rhinitis. AB - Indoor pollution is one of the most common problems addressed by allergists and troublesome for their patients. Although a large variety of products are available for removing such pollutants, including house dust, there is a relative paucity of data on the effectiveness of such devices. In many cases, central vacuum systems are recommended, particularly in new homes. To specifically address the question of whether a central vacuum system produces an improvement in the well characterized domains of Juniper Rhinoconjunctivitis Quality of Life Questionnaire, we selected 25 individuals with a history of documented type I hypersensitivity to house dust. Each of these individuals used either a Beam Central Vacuum System or their own conventional vacuum for a period of 3 months. At the end of this period, the individual switched over to the opposite limb of the study for 3 additional months. Interestingly, in all seven domains of the evaluation, including activity, sleep, nonnasal symptoms, practical problems, nasal symptoms, eye symptoms and emotions, use of the central vacuum proved to be superior. PMID- 11908817 TI - The role of R576Q polymorphism of interleukin-4 receptor alpha gene in atopy: results of a family-based study design. AB - INTRODUCTION: There is strong evidence that the R576 allele of interleukin-4 receptor alpha gene (IL-4R) might predispose to atopy. To test this hypothesis, we examined the association between the R576Q polymorphism and atopy in a Polish population using the family-based study design. MATERIALS AND METHODS: 44 atopic patients (age range from 11 to 34 years) with pollen and house dust mite allergy or/and mild asthma together with both parents were studied. The R576Q polymorphism of the IL-4R gene was genotyped in each patient and both parents, respectively, using the PCR-based protocol. The results were analyzed by the transmission disequilibrium test (TDT). The total IgE serum level, allergen specific IgE to the common aeroallergens, IL-4, and sIL-4Ralpha were assessed in each patient and both parents. RESULTS: In the TDT test the R576 and Q576 alleles were transmitted from the heterozygous parents to the affected offspring 20 and 15 times, respectively (McNemar test: p = 0.19). The results of the transmission disequilibrium test did not reach statistical significance. Thus, the R576 allele might contribute to the pathogenesis of allergic diseases in patients with high total IgE serum level (p < 0.05). A larger study group has to be studied to prove the observed linkage and association. PMID- 11908819 TI - Recurrent pneumonia as warning manifestation for suspecting primary immunodeficiencies in children. AB - Two hundred and eight children with recurrent pneumonia were studied over a 5 year period. Among these patients we found 10 cases with primary immunodeficiency disease: 6 cases of IgA deficiency, 1 case of X-linked agammaglobulinemia, 1 case of common variable immunodeficiency, 1 case of hyper IgM syndrome, and 1 case of Wiskott-Aldrich syndrome. This study describes the clinical features of these cases and assesses the usefulness of our immunodeficiency screening protocol. In this group 6 were males; the mean age at first episode of pneumonia was 3 years (range 3 months to 18 years), and the age of diagnosis ranged between 10 months and 19 years. The average number of episodes of pneumonia in each patient was 5 (range 2 to 12), and the number of hospitalizations ranged up to 13. The etiologic agents isolated from this recurrent pneumonia were S. pneumoniae, Moraxella, adenovirus, respiratory syncytial virus, and influenza B virus. Intravenous immunoglobulin was used in four cases. Two patients had chronic pulmonary damage with bronchiectasis and interstitial pneumonia. Only one patient died (Wiskott-Aldrich syndrome) during the follow-up from an intracranial hemorrhage. We found that the screening protocol applied to patients with recurrent pneumonia is a useful tool for ruling out the primary immunodeficiency disorders. PMID- 11908820 TI - Effects of long-term inhaled steroid use on bone mineral density in asthma patients. AB - Asthma is a chronic inflammatory disease of the airways. Inhaled corticosteroids are the most effective and most frequently used antiinflammatory drugs in the treatment of asthma. It is well known that long-term systemic steroid therapy causes osteoporosis, whereas it is thought that inhaled forms do not cause this side effect. Despite the recent disagreeing reports, it has been suggested that the use of inhaled steroids are associated with findings of biochemical bone changes. Therefore, we measured the bone mineral densities (BMD) of 18 patients (15 female, 3 male) with bronchial asthma receiving long-term inhaled steroids by X-ray absorptiometry technique and compared the results with those of 14 healthy control subjects (12 female, 2 male) who had been matched according to age, sex, menopausal state, and body mass index (BMI). RESULTS: There were no detectable significant difference between the BMD levels of left hip (trochanter major, neck of femur, Ward's triangle) and lumbar area of the spine (L2-L4) (p > 0.05). PMID- 11908821 TI - Long-term duration of reduced serum complement level following burn injury. AB - We report the case of a boy whose serum CH50 was below the detection limit following burn injury and skin transplantation. The APCH50 level was slightly decreased, although C3, C4, and the other complements were within the normal range. Cold activation was not detected in his plasma. His peripheral blood monocyte ratio slightly elevated to 19% and then decreased to 5.9%. In this case, burn injury caused the depletion of the complement, particularly in the alternative pathway, and resulted in the reduced CH50 level, although C3 did not show the typical pattern of alternative pathway depletion. In previously reported cases of burn injury, the CH50 level returned to the normal range within 2 weeks. In this patient the reduced level of CH50 continued for 4 months. We should consider burn injury one of the causes of complement deficiency even in cases with a duration of more than 1 month. PMID- 11908822 TI - Studies on functional roles of the histaminergic neuron system by using pharmacological agents, knockout mice and positron emission tomography. AB - Since one of us, Takehiko Watanabe (TW), elucidated the location and distribution of the histaminergic neuron system in the brain with antibody raised against L histidine decarboxylase (a histamine-forming enzyme, HDC) as a marker in 1984 and came to Tohoku University School of Medicine in Sendai, we have been collaborating on the functions of this neuron system by using pharmacological agents, knockout mice of the histamine-related genes, and, in some cases, positron emission tomography (PET). Many of our graduate students and colleagues have been actively involved in histamine research since 1985. Our extensive studies have clarified some of the functions of histamine neurons using methods from molecular techniques to non-invasive human PET imaging. Histamine neurons are involved in many brain functions, such as spontaneous locomotion, arousal in wake-sleep cycle, appetite control, seizures, learning and memory, aggressive behavior and emotion. Particularly, the histaminergic neuron system is one of the most important neuron systems to maintain and stimulate wakefulness. Histamine also functions as a bioprotection system against various noxious and unfavorable stimuli (for examples, convulsion, nociception, drug sensitization, ischemic lesions, and stress). Although activators of histamine neurons have not been clinically available until now, we would like to point out that the activation of the histaminergic neuron system is important to maintain mental health. Here, we summarize the newly-discovered functions of histamine neurons mainly on the basis of results from our research groups. PMID- 11908823 TI - Effects of a beta-adrenergic blocking agent timolol on intra ocular pressure responses induced by stimulation of cervical sympathetic nerve in the cat. AB - We clarified whether the intraocular pressure (IOP) response elicited by stimulation of the cervical sympathetic nerve (CSN) is influenced by changes in the baseline of IOP level and by beta-adrenergic blockade. The CSN was stimulated electrically for 30 seconds (10 V, 0.1-100 Hz, 2 milliseconds pulse duration) in urethane (100 mg/kg i.v.)-chloralose (50 mg/kg i.v.)-anesthetized, paralyzed cats. The IOP was monitored manometrically, and a controlled saline infusion was delivered into the anterior chamber to gradually increase IOP. CSN stimulation was delivered at the various baseline IOP levels so obtained. When required, a beta-adrenergic blocker timolol (2%) was delivered into the conjunctival cul-de sac. The normal IOP in our cats was 25+/-3 mmHg. This value decreased transiently on CSN stimulation. The amplitude of this IOP response depended on stimulus frequency and the pre-stimulus baseline IOP level. Topical administration of timolol increased the IOP response to CSN stimulation at a given baseline level. These results suggest that beta-adrenergic blockade increases the alpha adrenergic mediated-IOP reduction elicited by CSN stimulation at given baseline IOP level. PMID- 11908824 TI - Sampling methods and residential factors affecting formaldehyde concentration in indoor air. AB - Formaldehyde (HCHO) is the most serious residential pollutant. In order to evaluate residential HCHO levels, two sampling methods have been recommended; one is a 30 minute sampling in a closed room, and the other is a 24 hour sampling with an ordinary lifestyle routine. The aim of this report was to clarify the difference between the HCHO levels obtained by the two sampling methods. Residential air in 58 rooms was sampled for 30 minutes by an active sampling method more than 5 hours after residents closed windows, and by a passive sampling method for 24 hours with an ordinary lifestyle routine. The HCHO concentration with the 30 minute sampling was 0.118+/-0.065 ppm (range: 0.034 0.295 ppm) and 36 rooms (62%) exceeded the Japanese guideline value of 0.08 ppm, while 5% were higher than 0.25 ppm. The HCHO concentration with the 24 hours sampling was 0.053+/-0.039 ppm (range: 0.02-0.167 ppm) and only 13 rooms (22%) exceeded 0.08 ppm. The relationship between the concentrations obtained by the two methods was linear. However, the level with the 24 hour sampling significantly reduced with prolonged window opening time, meaning that occupants made an effort to reduce the usual exposure to about 40% of the exposure in a closed room by opening windows in order to escape from irritation. Since major adverse effects of HCHO are irritation and sensitization, the occasional peak concentration must be focused. In order to evaluate residential HCHO levels, measurement in a closed room is recommended even if people are living there. PMID- 11908825 TI - Different binding property of verotoxin-1 and verotoxin-2 against their glycolipid receptor, globotriaosylceramide. AB - We determined the binding of verotoxin-1 (VT1) and verotoxin-2 (VT2) against globotriaosylceramide (Gb3) by a monoclonal antibody (mAb)-based enzyme-linked immunosorbent assay (ELISA). Ethanolic solution of Gb3 containing cholesterol and phosphatidylcholine was passively adsorbed onto the wells of microtiter plate, and Gb3-bound VT1 and VT2 were detected by anti-VT1 and anti-VT2 mAbs, respectively. Although both VT1 and VT2 reacted with Gb3 in a concentration dependent manner, terminal galactose requirement for Gb3 binding was also different from each other. Pretreatment of VT1 showed the inhibitory effect on the binding of VT2 to Gb3, while the VT2-pretreatment showed no inhibitory effect on VT1 binding to Gb3. This was not due to the replacement of Gb3-bound VT2 with post-treated VT1. These results suggest that the binding sites of VT1 and VT2 on Gb3 are not identical to each other. PMID- 11908826 TI - Lung perfusion in hemorrhagic shock of rats: the effects of resuscitation with whole blood, saline or Hes 6%. AB - This study was undertaken to determine the effects of various resuscitation regimens on lung perfusion following resuscitation from hemorrhagic shock. Fourty male Sprague-Dawley rats (250-300 g) were used. The rats were divided randomly into four groups (n = 10 for each) and were sedated with intramuscular ketamine (100 mg/kg). We measured blood pressure, rectal temperature and lung perfusion using radioscintigraphy with a technetium colloid indicator. The systolic blood pressure was decreased 75% by removing blood via v. jugularis in the first three groups and group 4 was accepted as the control group, and blood volume was not diminished. Then the first three groups were resuscitated with autologous blood containing 125 units heparine/ml in group 1, saline in group 2, and hydroxyethyl starch (HES) 6% in group 3. After the correction of hypovolemia, all animals were injected 100 Bg (0.1 cc) technetium 99 m macroaggregated albumin (99mTc MAA) via penil vein. After injection of 99mTc MAA, 3 minutes fixed images were detected by a y camera in posterior position at 15 minutes and 5 hours. 99mTc MAA "wash out" rate in lung was determined quantitatively at 5 hours. Compared to a control group, lung perfusion was decreased significantly in groups resuscitated with saline, and HES 6% while perfusion was restored with autologous blood. We conclude that heparinized autologous blood saved lung capillary circulation in hemorrhagic shock in rats. PMID- 11908827 TI - Colposuspension as a treatment for urinary incontinence in spayed dogs. PMID- 11908828 TI - Mycoplasmal respiratory infections in small animals: 17 cases (1988-1999). AB - Seventeen cases (i.e., 14 dogs and three cats) were identified as having Mycoplasma spp. as the sole bacterial isolate cultured from airway washings in 224 cases evaluated for lower respiratory disease that was present in each case. Primary diagnoses included pneumonia (35.3%), airway collapse (35.3%), and bronchitis (29.4%). Fourteen cases had follow-up information available. Of these cases, eight showed resolution or improvement with antimycoplasmal drugs. Mycoplasma spp. is recognized as a primary cause of respiratory disease in several species, including humans. The relationship between Mycoplasma spp. and respiratory disease detected in some of these cases suggests some Mycoplasma spp. may act as primary pathogens in dogs and cats. PMID- 11908829 TI - Subcutaneous emphysema, pneumomediastinum, and pulmonary emphysema in a young schipperke. AB - A 4-month-old, intact female schipperke was presented for evaluation and treatment of subcutaneous (SC) emphysema. Radiographs revealed pneumomediastinum and SC emphysema. Sequential radiographs confirmed a worsening of the SC emphysema. Extensive, nonsurgical evaluation failed to reveal the source of the air within the mediastinum. Exploratory thoracotomy revealed an emphysematous right middle lung lobe. Lobectomy of the right middle lung lobe resolved both the pneumomediastinum and SC emphysema. Histopathological evaluation confirmed pulmonary emphysema. A variation of congenital pulmonary emphysema was considered in this case. PMID- 11908830 TI - Clinical signs, clinicopathological findings, etiology, and outcome associated with hemoptysis in dogs: 36 cases (1990-1999). AB - Hemoptysis, the expectoration of blood or bloody mucus from the respiratory tract at or below the larynx, was retrospectively evaluated in 36 dogs. Cough, tachypnea, and dyspnea were common historical and physical examination signs. Anemia was documented in 11 dogs, but was severe in only one dog. Other clinicopathological findings reflected the underlying diseases. All thoracic radiographs obtained were abnormal; alveolar and interstitial patterns were most common. Diseases predisposing to hemoptysis included bacterial bronchopneumonia (n=7), neoplasia (n=5), trauma (n=5), immune-mediated thrombocytopenia (n=4), heartworm disease (n=4), rodenticide poisoning (n=3), lung-lobe torsion (n=1), left-sided congestive heart failure (n=1), pulmonary hypertension (n=1), and foreign-body pneumonia (n=1). Four additional dogs had more than one underlying disease process. Nine dogs were either euthanized or died in the hospital during the initial visit. While at least half of the 27 dogs discharged went on to completely recover, five dogs discharged were known to have either died or been euthanized as a result of their disease in <6 months. PMID- 11908831 TI - Monoclonal gammopathies in the dog: a retrospective study of 18 cases (1986-1999) and literature review. AB - Eighteen dogs with monoclonal gammopathies were evaluated retrospectively. Most of the cases were associated with lymphoproliferative tumors (i.e., nine multiple myelomas, one Waldenstrom's macroglobulinemia, one lymphoma, one chronic lymphocytic leukemia, and one mucocutaneous plasmacytoma). The prevalence of nonmyelomatous monoclonal gammopathies (28%) was also significant (three leishmaniasis and two ehrlichiosis). Presenting complaints and clinical signs often were nonspecific or related to bleeding diathesis. Significant laboratory findings included proteinuria, hypoalbuminemia, and anemia. Some unusual features were also observed: a multiple myeloma with immunoglobulin M secretion, another myeloma with two narrow spikes on the electrophoretic pattern, and a mucocutaneous plasmacytoma secreting an immunoglobulin G paraprotein. PMID- 11908832 TI - Suspected paraspinal abscess and spinal epidural empyema in a dog. AB - Epidural spinal cord compression was visualized myelographically in a dog presented for rapid development of paraparesis. A large, fluid-filled pocket in the epaxial musculature was found at surgery and appeared to communicate with the first lumbar vertebra. Unfortunately, cytopathological evaluation of the fluid was not performed. No etiological agents were isolated on aerobic culture. The dog responded well to decompressive surgery and medical therapy consisting of antibiotics, pain medication, and nursing care. In the veterinary literature, only two studies of spinal epidural empyema in the dog have been reported. Of these dogs, one had successful decompressive surgery performed. The other dogs in these two reports were euthanized. The dog presented in this report fully recovered. Spinal epidural empyema should be considered as a differential diagnosis in dogs presenting with a fever and a rapidly progressing myelopathy. PMID- 11908833 TI - Tetraparesis in a cat with fibrocartilaginous emboli. AB - An 8-year-old cat, with a history of ataxia that progressed to tetraparesis over a 5-day period, was evaluated. A lesion was localized to the sixth cervical (C6) to second thoracic (T2) spinal cord segments based on physical and neurological examination findings. Blood work was unremarkable, as was survey radiography of the thoracic and abdominal cavities. Cerebrospinal fluid analysis showed moderate neutrophilic inflammation. A definitive diagnosis was not made until necropsy, at which time intravascular fibrocartilaginous embolization (FCE) of the cervical spinal cord was identified. This is only the third published report of FCE in the feline species and the first such case involving the cervical spinal cord. PMID- 11908834 TI - Canine gastric adenocarcinoma and leiomyosarcoma: a retrospective study of 21 cases (1986-1999) and literature review. AB - This retrospective study describes the clinical course, treatment, and outcome of 21 dogs with gastric adenocarcinomas (n=19) and leiomyosarcomas (n=2). Medical records from 1986 to 1999 were reviewed for signalment, weight, diagnosis, tumor location, clinical signs, radiographic imaging procedures, surgical procedures, chemotherapy, duration of follow-up monitoring, outcome, cause of death, metastatic rate, metastatic sites, and method of detection of metastasis. Fourteen of 19 (74%) dogs with gastric adenocarcinomas had metastasis. Metastatic sites included gastric lymph nodes, omentum, liver, duodenum, pancreas, spleen, esophagus, adrenal glands, and lungs. Both cases of a gastric leiomyosarcoma had metastatic disease involving the liver (n=2) and duodenum (n=1). Surgery, consisting of either a Billroth I or a gastrojejunostomy, provided immediate relief of the gastric outflow obstruction and clinical improvement in the early postoperative period. The beneficial effects of chemotherapy alone or adjuvant chemotherapy are still unknown. Recurrence of clinical signs 3 days to 10 months after surgery caused all owners to elect euthanasia. The long-term prognosis for most cases of gastric adenocarcinomas and leiomyosarcomas is poor because of the presence of advanced disease. Surgical resection, however, does alleviate gastric outflow obstruction in the immediate postoperative period. PMID- 11908835 TI - Synovial T-cell lymphoma of the stifle in a dog. AB - A 6-year-old, 43-kg, spayed female rottweiler was presented for a 1-month history of progressive, left hind-limb lameness. Upon physical examination, a cranial drawer sign and joint distention were present in the left stifle. Radiographically, the stifle had evidence of effusion, remodeling of the patella, and an enlarged popliteal lymph node. Marked synovial thickening and an intact cranial cruciate ligament were noted during surgery. Despite finding a nonspecific, mixed inflammatory response on joint fluid cytopathology, histopathology demonstrated T-cell lymphoma of the synovium. Lameness may be the sole presenting clinical sign in canine lymphoma. PMID- 11908836 TI - Arthroscopic biceps brachii tenotomy as a treatment for canine bicipital tenosynovitis. AB - Five dogs of varying breeds, ranging from 4 to 8 years in age, were presented with clinical signs consistent with bicipital tenosynovitis. After failure of conservative treatment, each dog underwent shoulder arthroscopy. Following examination of the scapular humeral joint, the bicipital tendon was severed with a bipolar radiofrequency electrosurgical system. The arthroscopic procedure resulted in a good to excellent outcome for all five dogs. PMID- 11908837 TI - Spontaneous gastroduodenal perforation in 16 dogs and seven cats (1982-1999). AB - The records of 23 dogs and cats diagnosed with spontaneous gastroduodenal perforation (GDP) were retrospectively reviewed. Survival was 63% in dogs and 14% in cats. Rottweilers <5 years of age were overrepresented. Clinical evidence of gastrointestinal bleeding was common in dogs but not in cats. Shock was an uncommon presenting condition in dogs and was not closely linked to outcome. In fact, progression of an ulcerating lesion to GDP was not associated with marked changes in symptoms exhibited by many patients in this study. Most GDPs were associated with histopathological evidence of subacute or chronic peritoneal reaction at the time of diagnosis. This suggests that diagnostic methods employed lacked sensitivity in identifying early perforating lesions, and that dramatic signs of acute abdomen following gastroduodenal perforation may not be as common as was previously thought. PMID- 11908838 TI - Traumatic rupture of the ureter: 10 cases. AB - A retrospective study was performed on eight dogs, one cat, and one ferret with ruptured ureters secondary to blunt trauma. The most common physical examination findings were abdominal distension/discomfort (in five of 10 animals) and gross hematuria (in five of six animals). Multiple organ injury was also common (in seven of 10 animals). Loss of retroperitoneal and peritoneal detail was the most common radiographic finding (in four of six animals). Ureteronephrectomy was the most common surgical procedure (performed in five out of seven procedures). Three of the five cases discharged were available for follow-up and have had no evidence of associated problems. PMID- 11908839 TI - A stochastic model of multistable visual perception. AB - Multistability in vision is an intriguing phenomenon that is currently not well understood. In this paper, we present a new, stochastic model for multistable visual perception. It is based on results of time series analysis of experimental data, yielding evidence for it being a linear, stochastic process. This is the outcome of testing for unstable periodic orbits and comparing the correlation dimension of the data to that of white noise. In the model, all degrees of freedom but one can be determined by general knowledge, thus resulting in a high degree of parsimony. The remaining parameter is used to model the individual characteristics that vary between subjects. Fitting simulations to the experimental data proves the parameter to be in a physiologically highly plausible range. PMID- 11908840 TI - Changes in the fluctuation of interbeat intervals in spontaneously beating cultured cardiac myocytes: experimental and modeling studies. AB - Isolated and cultured neonatal cardiac myocytes contract spontaneously and cyclically, and have the properties of a non-linear oscillator. In this study, we have analyzed the relationship between the fluctuation of contraction rhythm of spontaneously beating cultured cardiac myocytes, and the coupling strength among them. The coefficient of variation of contraction intervals increased transiently in the early stages of incubation, and then decreased almost monotonically with time. The contraction rhythm of the myocytes became synchronized in the late stage of the culture. The day on which synchronization occurred almost coincided with the day when the coefficient of variation reached its lowest value. In addition, we have performed a mathematical analysis using interacting Bonhoeffer van der Pol oscillators to clarify the mechanisms underlying the changes in the fluctuation of contraction rhythm with time. As the coupling strength among oscillators increased, the coefficient of variation of oscillation periods increased temporarily, but then decreased rapidly when the oscillators showed synchronization. These results suggest that the changes in the fluctuation of beating rhythm result from the increase in strength of electrical coupling among spontaneously beating cardiac myocytes. PMID- 11908841 TI - Response of a pacemaker neuron model to stochastic pulse trains. AB - We investigated the response of a pacemaker neuron model to trains of inhibitory stochastic impulsive perturbations. The model captures the essential aspect of the dynamics of pacemaker neurons. Especially, the model reproduces linearization by stochastic pulse trains, that is, the disappearance of the paradoxical segments in which the output firing rate of pacemaker neurons increases with inhibition rate, as the coefficient of variation of the input pulse train increases. To study the response of the model to stochastic pulse trains, we use a Markov operator governing the phase transition. We show how linearization occurs based on the spectral analysis of the Markov operator. Moreover, using Lyapunov exponents, we show that variable inputs evoke reliable firing, even in situations where periodic stimulation with the same mean rate does not. PMID- 11908842 TI - Relevant EEG features for the classification of spontaneous motor-related tasks. AB - There is a growing interest in the use of physiological signals for communication and operation of devices for the severely motor disabled as well as for healthy people. A few groups around the world have developed brain-computer interfaces (BCIs) that rely upon the recognition of motor-related tasks (i.e., imagination of movements) from on-line EEG signals. In this paper we seek to find and analyze the set of relevant EEG features that best differentiate spontaneous motor related mental tasks from each other. This study empirically demonstrates the benefits of heuristic feature selection methods for EEG-based classification of mental tasks. In particular, it is shown that the classifier performance improves for all the considered subjects with only a small proportion of features. Thus, the use of just those relevant features increases the efficiency of the brain interfaces and, most importantly, enables a greater level of adaptation of the personal BCI to the individual user. PMID- 11908843 TI - Neural network simulations of the primate oculomotor system IV. A distributed bilateral stochastic model of the neural integrator of the vertical saccadic system. AB - The present report examines the performance of a distributed bi-directional neural network that simulates the vertical velocity to position integrator of the primate brain. Consistent with anatomy and physiology, its units receive stochastically weighted input from vertical medium-lead burst neurons. Also consistent with anatomy, units belonging to integrators with opposite on directions (up or down) are interconnected via the posterior commissure (again in a stochastically weighted manner) and they can be excitatory or inhibitory. To demonstrate that integration can be a one-step process, the output of model units was routed directly to vertical motoneurons. Model units replicate the wide range of saccade-related discharge patterns encountered in the portion of the primate brain that is thought to house the vertical neural integrator (the interstitial nucleus of Cajal) while "lesions" of model units and/or their interconnections replicate the symptoms which follow insults to this brain area. PMID- 11908844 TI - Differences in automatic social information processing between nondepressed and subclinically depressed individuals. AB - The present research examined individual differences in automatic social information processing. We hypothesized that because nondepressed and subclinically depressed persons have different interpersonal experiences, they may process social information in different ways. In this experiment, participants were asked to make judgments about social relationships after being reminded of a target person. They had to make these judgments under either a light or a heavy memory load. Results showed that when nondepressed participants were reminded of people with whom they had frequent pleasant interactions, they made a greater number of positive judgments about their social relationships than did subclinically depressed participants. When subclinically depressed participants were reminded of people with whom they had had frequent unpleasant interactions, they made a greater number of negative judgments about their social relationships than did their nondepressed counterparts. Moreover, performance in these experimental conditions was unaffected by memory load, suggesting that automatic thoughts about their social relationships had been evoked. PMID- 11908845 TI - General and specific traits of personality and their relation to sleep and academic performance. AB - Few studies have examined the links between personality variables and sleep and their combined effect on specific real-world outcomes. Participants in this study completed numerous personality, sleep, and performance measures; we examined the associations among these measures. Personality was assessed using the Five-Factor Model. The personality trait of Conscientiousness (especially its facet of Achievement Striving) was a substantial predictor of academic performance. Analyses of the sleep variables revealed three distinct constructs: quantity, quality, and schedule. Sleep quantity showed few interesting correlates. In contrast, sleep quality was associated with greater well-being and improved psychological functioning, whereas sleep schedule (i.e., average rising and retiring times) was significantly related to Conscientiousness, such that conscientious individuals maintain earlier schedules. PMID- 11908846 TI - Hope, defense mechanisms, and adjustment: implications for false hope and defensive hopelessness. AB - Two studies replicated and expanded an earlier finding that defense style plays a crucial role in the relation between hope and dysphoria (Kwon, 2000). Lower hope and higher defense style immaturity were each associated with greater dysphoria, depression proneness, and maladjustment. Individuals with low hope and low defense immaturity did not have poor outcomes, supporting the existence of a subtype of low hope (defensive hopelessness) that may have adaptive value. The combination of high hope and high defense immaturity was not associated with maladaptive outcomes, arguing against the false hope construct. Additionally, the findings remained after controlling for levels of anxiety. Thus, it appears that the results are not attributable to general distress or negative affectivity. Finally, domain-specific hope was shown to correlate most strongly with matching areas of adjustment, providing evidence for the validity of the construct. PMID- 11908847 TI - Day-to-day relationships between self-awareness, daily events, and anxiety. AB - Every day for 3 weeks, 41 participants provided measures of their state private and public self-consciousness (self-awareness, SA), and anxiety, and they described the events that occurred each day. Multilevel random coefficient modeling analyses found that daily private and public SA were positively related to the importance and frequency of daily negative social events and to daily anxiety. Public SA was also positively related to the importance and frequency of daily positive social events. Neither public nor private SA was related to the importance and frequency of daily achievement events. The strength of the relationship between public SA and positive social events was stronger for people who were less anxious, less depressed, and for those with greater self-esteem. Analyses of lagged relationships suggested that increased private SA led to increased negativity of social events, whereas increased public SA led to increased positivity of social events, and increased anxiety led to increased private SA. PMID- 11908848 TI - A model for the thermal unfolding of amicyanin. AB - In the present study the thermal unfolding of amicyanin has been addressed using differential scanning calorimetry, fluorescence emission, optical density, circular dichroism and electron paramagnetic resonance. The combined use of these techniques has allowed us to assess, during unfolding of the protein, its global conformational changes in relationship to the local structural modifications occurring in the copper environment and close to the fluorescent chromophore Trp46 of the protein. The thermal transition from the native to the denatured state is on the whole irreversible and occurs in the temperature range between 65 and 72 degrees C, depending on the scan rate and technique used. Amicyanin as a whole shows a complex unfolding pathway, which has been described in terms of a three-step model: N <--> U --> F1 --> F2. According to this model, in the first step the native state of the protein (N) goes reversibly to the unfolded state (U), in the second one U goes irreversibly to F1 and, finally, the state F2 is irreversibly reached in the third step. Kinetic factors prevent the experimental separation of these steps. Nevertheless, the comparison of the data obtained with the different experimental techniques testifies the presence, within the unfolding pathway, of some intermediate states, although not sufficiently long lived to allow a detailed characterization. A first intermediate transient state has been identified around 68 degrees C, whereas a second one can be related to conformational changes that involve the copper environment. Finally, an exothermal phenomenon, caused by irreversible rearrangements of the melted polypeptide chains, is evidenced. In addition, according to the EPR findings, the type 1 copper ion, which is four-fold coordinated by two N and two S atoms in a distorted tetrahedron in the native state of the protein, shows type 2 features after denaturation. A mathematical model simulating the unfolding Cp(exc) profile has been also developed. PMID- 11908849 TI - Two-photon excitation and photolysis by pulsed laser illumination modelled by spatially non-uniform reactions with simultaneous diffusion. AB - Two-photon absorption in the focus of a pulsed laser has the potential for localized photolysis of caged compounds, generating high concentrations of neurotransmitters, hormones and messengers. The concentrations of cage, intermediates and products in the femtolitre focal volume depend on reaction rates and diffusional exchange with the external volume. This problem of reaction with diffusion was analysed with analytical and numerical methods to determine simple relations between parameters useful in the design and interpretation of experiments. The diffraction-limited laser spot is approximated well by a sphere, radius A, in diffusional exchange with either an infinite uniform medium, representing extracellular photolysis, or within a non-permeable sphere, a "cell" of radius B, representing intracellular photolysis. Photolysis is modelled as sequential irreversible reactions, with either the excitation step alone, rate constant k(e), or with a subsequent "dark" reaction, rate constant k(p). For extracellular photolysis, steady-state depletion of a cage averaged in a spherical spot increases hyperbolically with k(e) with half-maximum depletion at k(e) = K0.5 = 2.5 D/A2, where D is the diffusion coefficient. With measured parameters for spot size A = 0.3 microm and diffusion D = 800 microm2/s, K0.5 = 22,200 s(-1). The optimal exposure for localized photolysis is the characteristic diffusion time tau = A2/D, 113 micros in this example, and is the time taken to reach 57% of steady state in the diffusion-limited case. In the two-step model, with excitation and "dark" reaction steps, rate constants both exceeding K0.5 are necessary to generate 50% of maximal product concentration in the illuminated volume. High concentrations of photolysis products depend particularly on a high excitation rate constant (k(e) > K0.5), and localization of the products requires fast dark reactions (k(p) > K0.5). If products diffuse faster than the cage, their steady-state concentrations are decreased, and concentration transients may occur. For localized intracellular photolysis, the duration of exposure that generates product concentration at the cell boundary, B, less than 10% of the spot concentration should be shorter than 0.043(B/A)3tau, and is determined by diffusion. PMID- 11908850 TI - The efficiency of two-photon photolysis of a "caged" fluorophore, o-1-(2 nitrophenyl)ethylpyranine, in relation to photodamage of synaptic terminals. AB - Localized photolysis of caged neurotransmitters with the two-photon effect for investigations at synaptic preparations was evaluated by determining the toxicity to synaptic transmission of pulsed near-IR laser light focused into the terminals of the snake neuromuscular junction, and measuring the extent of photolysis of a conventional caging group with similar irradiation in microcuvette experiments. Photodamage was seen in synaptic terminals as a large, irreversible increase of spontaneous synaptic activity with laser flashes of 5 ms at 1 Hz at average powers > 5 mW and was due to multiphoton absorption. Localized photolysis due to two-photon absorption was investigated for a representative caged fluorophore, the 1-(2-nitrophenyl)ethyl ether of pyranine (NPE-HPTS). Irradiation of NPE-HPTS at 5 mW with the same optical arrangement produced very low rates of photolysis. NPE-HPTS photolysis mechanisms were investigated at high laser powers by measuring (1) the kinetics of two-photon fluorescence generated by two-photon photolysis in the focal volume and (2) the rates of HPTS accumulation inside closed 2-10 microm radius vesicles, measured with one-photon excitation during two-photon photolysis by repetitive 10 micros laser exposures. The two-photon crosssection of NPE-HPTS photolysis calculated from the rates is 0.02-0.04 GM (10(-50) cm4 x s/photon) and limits the efficiency of photolysis at 5 mW. With free diffusional exchange, 50% steady-state cage depletion in the focal volume was estimated to occur only at high laser powers of ca. 72 mW, masked in experiments by multiphoton bleaching. Based on these results, the two-photon photolysis cross-section needed for 50% steady-state photolysis of a caged neurotransmitter at 5 mW is calculated as 31 GM, much higher than in existing caged compounds. PMID- 11908851 TI - Shear stress induces caveolin-1 translocation in cultured endothelial cells. AB - Considering that vascular endothelial caveolae could be flow sensors converting mechanical stimuli into chemical signals transmitted into the cell, this work studied, in vitro, the change of caveolin-1 expression and distribution of cultured endothelial cells exposed to laminar flows. Experimental results showed that, in control cells, caveolin-1 were primarily localized on the cell surface, and presented some local concentrations. In cells exposed to laminar flows, caveolin-1 distribution showed a time-dependent variation. After 24 h of shear (1.0 Pa), the expression of caveolin-1 increased and a local caveolin-1 concentration was found, in most cells, at the upstream side of the cell body where the hydrostatic pressure and the spatial gradient of shear stress were at a maximum. As a comparison, tumor necrosis factor-a induced a decrease of caveolin 1 in the cells. PMID- 11908852 TI - Spectroscopic evidence for site-specific cellular activity in the tubular gland in human intestine. AB - The intestinal crypts contain mucus-secreting goblet cells in large numbers. In the tubular gland (crypt), the cells are generated at the bottom and end their life cycle at the top. Recently, FTIR microspectroscopy (FTIR-MC) has been applied in biology and medicine. The characterization of various cellular types using FTIR-MC and its subsequent application for the diagnosis of cancer is becoming a reality. In this report, we investigate the differential cellular activity in the normal tubular gland using FTIR-MC. Our results indicate that the absorbance for the cells in the bottom of the crypt is always higher than those in the upper portion. There are spectral pattern changes and frequency shifts for cells at the bottom and top sites of the normal crypt. Also, the comparison of a normal crypt with a malignant one has been made. This is the first spectroscopic evidence in the literature showing the difference in the cellular activity at different sites in the tubular gland. The reasons for our observations and their implications are discussed. PMID- 11908853 TI - Evidence for direct interaction between actin and the cystic fibrosis transmembrane conductance regulator. AB - Previous studies have demonstrated that actin filament organization controls the cystic fibrosis transmembrane conductance regulator (CFTR) ion channel function. The precise molecular nature of the interaction between actin and CFTR, however, remains largely unknown. In this report, interactions between actin and purified human epithelial CFTR were directly assessed by reconstitution of the channel protein in a lipid bilayer system and by atomic force microscopy (AFM). CFTR containing liposomes in solution were deposited on freshly cleaved mica and imaging was performed in tapping-mode AFM. CFTR function was also determined in identical preparations. Images of single CFTR molecules were obtained, and addition of monomeric actin below its critical concentration showed the formation of actin filaments associated with CFTR. The data indicate a direct interaction between actin and CFTR exists, which may explain the regulatory role of the cytoskeleton in ion channel function. This was confirmed by functional studies of CFTR single-channel currents, which were regulated by addition of various conformations of actin. The present study indicates that CFTR may directly bind actin and that this interaction helps affect the functional properties of this channel protein. PMID- 11908854 TI - Effect of strand orientation on the interaction of berenil with DNA triple helices. AB - Using circular dichroism spectroscopy the ability of berenil, a minor groove binding drug, to induce triple helix formation was investigated with two oligonucleotides designed to form two intramolecular triplexes containing T*A:T and G*G:C triplets, which differ only by the orientation of their third strand: 5'-d(G4A4G4-[T4]-C4T4C4-[T4]-G4T4G4), and 5'-d(G4T4G4-[T4]-G4A4G4-[T4]-C4T4C4), where [T4] represents a stretch of four thymine residues. We demonstrate that when added to the duplex form of these oligonucleotides, berenil induces triplex structure formation only if the orientation of third strand is anti-parallel to the purine strand. PMID- 11908855 TI - Risk factors for methicillin-resistance among patients with Staphylococcus aureus bacteremia at the time of hospital admission. AB - BACKGROUND: Community-acquired infections caused by methicillin-resistant Staphylococcus aureus (MRSA) seem to be increasing. Characteristics permitting recognition of patients with such strains would aid infection control efforts and choice of empiric therapy pending culture and susceptibility results. METHODS: Retrospective review of medical records for all adults seen in the Emergency Care Center at Grady Memorial Hospital, Atlanta, Georgia, whose blood cultures taken within 24 hours of entry yielded S. aureus. Risk factors for the presence of methicillin resistance in S. aureus isolates recovered from patients with staphylococcal bacteremia were assessed. RESULTS: S. aureus isolates from 118 (40%) of 297 study patients with bacteremia at the time of admission were methicillin-resistant. Multivariate analysis identified hospitalization in the 6 months preceding admission [odds ratio (OR) = 4.4; 95% CI, 2.0-9.8], receipt of antimicrobial agents in the past 3 months (OR = 5.6; 95% CI, 2.6-11.9], presence of indwelling urinary catheter (OR = 7.3; CI, 2.5-20.9), and nursing home residence (OR = 9.9; 95% CI, 3.9-25.6) to be independently associated with the presence of methicillin resistance. All but 4 of the 118 patients with methicillin-resistant strains had at least 1 of these factors and the proportion of resistant isolates progressively increased as more of these features were present. CONCLUSIONS: The presence of these risk factors should be considered when making decisions about isolation and other infection control procedures as well as empiric antimicrobial therapy with vancomycin for patients with suspected staphylococcal infection at the time of hospital admission. Similar studies could guide practices for dealing with such patients in other centers, because the occurrence of MRSA infections at the time of admission may vary widely by geographic area. PMID- 11908856 TI - Iterative method for learning skills as an efficient outpatient teacher. AB - BACKGROUND: We sought to identity the choices and the methods used by ambulatory teachers in a qualitative study, using teacher-intern-patient role plays to improve ambulatory teaching. METHODS: We used repeated performances of a scripted role play; during each iteration, field notes were taken by the authors. Insights garnered at each iteration were incorporated into the next version of the role play. After 9 iterations, no further insights into outpatient teaching were forthcoming, and our observations were included into a qualitative study. RESULTS: The sequence of steps and major choices to be made in an outpatient teaching encounter were delineated. The goals of the initial opening phase were defined as setting a learning climate, gathering information about the case, and assessing the learner's level of knowledge. Alternatives posed for setting up the second phase of the interaction with the patient included the choice of being a role model or being a "coach." Three-way conversations between patient, learner, and teacher were described in this phase of the encounter. In the final or summary phase of the encounter, we described the choices between giving a "general rule" for learning, and/or exploring higher-level issues, such as patient-doctor communication skills, medical ethics, or feedback for the learner. CONCLUSIONS: The sequence of steps involved in an outpatient teaching encounter, were defined, and the major choices to be made in the encounter were described. PMID- 11908857 TI - Diffuse pulmonary hemorrhage: evolutionary 'flaw' or consequence of evolutionary progress? AB - Diffuse alveolar hemorrhage (DAH), a serious and frequently life-threatening process, can occur in a wide variety of disorders, including mitral stenosis, systemic autoimmune diseases, conditions associated with anti-neutrophil cytoplasmic antibodies, anti-glomerular basement membrane antibody, inhaled toxins or infection, and drug allergies. These causes have no common denominator, but the end result, of capillary inflammation and hemorrhage, is the same. The capillaritis seen in DAH is present exclusively in the lung and is not part of a more generalized systemic vasculitis. In a recent article, I highlighted the central role of the neutrophils in this process, and suggested that in DAH the neutrophils present in the lung may sift the associated immune complexes and transfer them out of the vascular system into the alveoli. In this article, we document the inflammation seen during the remission period and during the acute lung bleeding of DAH in 2 patients. Because DAH is a common finding in a variety of pathological conditions, we then explore the possible evolutionary mechanisms behind DAH. PMID- 11908858 TI - Insulin: a novel factor in carcinogenesis. AB - Cancer is a leading cause of mortality in the United States. Despite much research on specific carcinogens, the cause of many cancers remains unclear. The identification of novel causative agents offers the potential for cancer prevention. Diseases such as obesity and diabetes mellitus, characterized by hyperinsulinemia, are associated with increased risk of endometrial, colorectal, and breast carcinomas. There is increasing evidence that insulin is a growth factor for tumor formation. The mechanisms underlying insulin-mediated neoplasia may include enhanced DNA synthesis with resultant tumor cell growth, inhibition of apoptosis, and altered sex hormone milieu. The reduced insulin levels seen with physical activity, weight loss, and a high fiber diet may account for decreased cancer risk. The role of newer drugs that restore sensitivity to insulin, thereby reducing hyperinsulinemia, is an exciting potential area of cancer prevention. In this review, we discuss the potential role of insulin as a tumor growth factor. PMID- 11908859 TI - Case records of the VA Maryland Healthcare System/University of Maryland Medicine. A 75-year-old man with right upper quadrant pain and gallstones. PMID- 11908860 TI - Pulmonary artery thrombus and subcapsular liver hematoma in a patient with HELLP syndrome: a therapeutic conundrum. AB - HELLP syndrome (hemolysis, elevation of liver enzymes, and low platelet count) occurs during pregnancy. Intrahepatic hemorrhage and subcapsular liver hematoma with or without rupture are reported complications of this syndrome. The patient described in this report developed HELLP syndrome associated with a subcapsular liver hematoma and pulmonary artery thrombus, complications that created a therapeutic conundrum. PMID- 11908861 TI - Tumor lysis syndrome after hydrocortisone treatment in metastatic melanoma: a case report and review of the literature. AB - Tumor lysis syndrome (TLS) is 1 of the complications that usually follows chemotherapy treatment of myelo-lymphoproliferative diseases. Corticosteroids (CS) could also induce TLS in this type of malignancies. On the other hand, TLS in solid tumors is less frequent, and CS treatment was never reported to be associated with TLS in solid tumor. Here we report the first case of TLS in a solid tumor (melanoma) after CS treatment. PMID- 11908862 TI - Pleuritis as a presenting manifestation of rheumatoid arthritis: diagnostic clues in pleural fluid cytology. AB - Pleural effusion is a recognized but relatively uncommon complication of rheumatoid arthritis and has distinctive cytopathologic features. It may occur before, concurrently with, or after the development of joint manifestations of the disease. Clinical diagnosis of rheumatoid pleuritis may be delayed or overlooked in a patient without obvious evidence of arthritis. Failure to recognize the unique cytologic picture of rheumatoid pleuritis caused a 5-month delay in diagnosis of a rheumatoid pleural effusion in an elderly man whose concurrent arthritic symptoms were not given adequate clinical recognition. PMID- 11908864 TI - Acupuncture therapy for persistent hiccups. AB - Persistent hiccups (singultus) is a rare but severely disabling disorder. The causes of persistent hiccups are numerous, as are the treatment options. However, none of the treatment modalities has proven to be effective by evidence-based criteria, and no treatment has been shown to be superior to another. Traditional acupuncture has not been previously reported as a modality for the treatment of persistent hiccups in the English medical literature. We describe 2 patients with persistent hiccups refractory to conventional treatments that were treated successfully using acupuncture. PMID- 11908863 TI - Transient Graves disease developing after surgery for Cushing disease. AB - A 49-year-old man, diagnosed as having Cushing disease in 1976 at the age of 26, underwent a Hardy operation 13 years after treatment with reserpine combined with pituitary radiation. In laboratory examinations before and 2 weeks after the successful surgery, the patient's serum thyroid hormones were found to be normal except for suppressed serum thyroid-stimulating hormone (TSH), and his serum anti TSH receptor (TRAb) and anti-TSH receptor-stimulating antibodies (TSAb) were negative. Glucocorticoid supplemental treatment was withdrawn on the 15th day after surgery and was restarted on the 48th day, during which time there were no signs of an adrenal crisis. Sinus tachycardia, fine finger tremor, and enlarged thyroid gland, approximately the size of a thumb head, were observed on the 140th day after surgery. Thyrotoxicosis with increased serum TSAb and TRAb and high 24 h thyroid uptake of 123I was noted, indicating a diagnosis of Graves disease. No special treatment was prescribed, but his serum thyroid hormone levels started to decrease on the 140th day after the operation and returned to normal on the 520th day. Serum TRAb also spontaneously decreased, but the timing of the peak of serum TRAb was delayed 230 days from that of the thyroid hormones. This is the first reported case of Graves disease after successful surgery for Cushing disease. We presume that a latent autoimmune process in the thyroid, suppressed by hypercortisolism, developed into overt Graves disease after the abrupt reduction of plasma glucocorticoid levels induced by surgery. PMID- 11908865 TI - Primary lymphoma of the central nervous system: a clinicopathologic study. AB - We performed clinicopathologic examinations of 27 cases of primary lymphoma of the central nervous system not related to acquired immune deficiency syndrome. We considered age and change of performance status (PS) to be especially important in clinical examination. We also conducted pathological studies of these tumors and the characteristics of their cells, in order to characterize pathological subtypes, cell kinetics, and involvement of viruses. PS of patients more than 70 years old decreased markedly before treatment and did not show the improvement after treatment that was exhibited by those under 70 years of age. Low PS (60% or less) after initial treatment, high MIB-1 positivity (over 44.0%), and high counts of AgNOR (over 4.56/cell) were significantly associated with lower survival rates. Patients with immunoblastic lymphoma and high MIB-1 positivity are likely to die from general debilitation, without evidence of recurrence from imaging. Preoperative steroid therapy was significantly associated with higher apoptotic positivity. PMID- 11908866 TI - Histone deacetylase inhibitors such as sodium butyrate and trichostatin A induce apoptosis through an increase of the bcl-2-related protein Bad. AB - The effects of sodium butyrate (SB) and trichostatin A (TSA) on cell proliferation andapoptosis against human glioma T98G, U251MG, and U877MG cells were investigated. Upon exposure to either SB or TSA, cell proliferation was reduced, and apoptosis detected by DNA fragmentation analysis and the cleavage of CPP32 was induced. Previously, we reported that SB increased the expression levels of p21 (WAF-1) and inhibited G1-S transition of the cell cycle. In this study, we showed that TSA also increased p21 expression, suggesting that histone deacetylase (HDAC) inhibitors may up-regulate p21 protein in common and thus arrest proliferation in the G1 phase of the cell cycle. To further determine the underlying molecular mechanisms of apoptosis with either SB or TSA treatment, we studied the expression levels of apoptosis-related proteins in human glioma cells. SB increased the expression of the Bad protein, although the expression of Bcl-2, Bcl-xL, Bax, and Fas was not changed by theaddition of SB. TSA treatment also up-regulated the expression of Bad protein. The results suggest that HDAC inhibitors such as SB and TSA induce apoptosis through an increase in Bad protein in human glioma cells in vitro. PMID- 11908867 TI - Morphologic features of human chorionic gonadotropin- or alpha-fetoprotein producing germ cell tumors of the central nervous system: histological heterogeneity and surgical meaning. AB - Our study of germ cell tumors (GCT) of the central nervous system (CNS) investigated the relationship between tumor histology and patient serum titers of human chorionic gonadotropin (HGC) and alpha-fetoprotein (AFP). Thirty-five patients were enrolled. Their serum titers of HCG (mlU/ml) and/or AFP (ng/ml) before initial treatment were available, as were tumor specimens obtained before the administration of adjuvant therapy. They were divided into three groups, depending on whether HCG alone (group H), AFP alone (group A), or both HCG and AFP (group HA) were detected. Each group was subdivided into three groups: patients in group I had H, A, and/or HA titers below 9.9; patients in group II/III had titers from 10.0 to 999; and those in group IV had titers of 1000 or more. Serial sections of tissue specimens were repeatedly stained, mainly with hematoxylin and eosin (H-E) stain, HCG immunostain, and AFP immunostain. There were seven patients in the H-I group and five in H-II/III. Of these 12 patients, 11 had germinomas (G) and one had an embryonal carcinoma (EC). Five patients were included in group A: one was classified as A-II/III and had a germinoma, and the remaining four patients were in A-IV and had yolk sac tumors (YST) or mixed GCT consisting mainly of YST or EC (MXGCT-YST, EC). The HA group consisted of 18 patients. Three were classified as HA-I and had germinomas; nine HA-II/III patients had T or MXGCT-T; and six HA-IV patients had choriocarcinoma (CC), YST, MXGCT-CC, or MXGCT-YST. Throughout the study, the situations for the elevated serum titers could be elucidated in only four cases (three in group A-IV and one in group HA-IV). These results led to the conclusion that serologic evaluation is superior to morphologic evaluation in diagnosing marker-producing GCTs. From a diagnostic perspective, the role of surgery is to verify the HCG- and AFP immunonegative tissue in patients with G, T, and EC. PMID- 11908868 TI - Expression of Bcl-2, Bcl-x, and Bax proteins in astrocytomas in relation to patient survival. AB - The Bcl-2 family is composed of a group of related proteins that either prevent or promote apoptosis. This study was undertaken to assess the prognostic value of Bcl-2, Bcl-x, and Bax in patients with astrocytomas. Tissue samples from 104 astrocytomas (WHO grade 2, 21 cases: grade 3, 49 cases; grade 4, 34 cases), including 68 primary and 36 recurrent tumors, were examined immunohistochemically for Bcl-2, Bcl-x, and Bax expression. Patient charts were reviewed for clinical presentation, and survival was followed. The mean values of the Bcl-2, Bcl-x, and Bax labeling indexes (LI) were 15.9 +/- 13.1%, 53.2 +/- 35.3%, and 25.9 +/- 23.2%, respectively. Statistical analysis showed that the Bcl-x LI of high-grade (grade 3 or 4) astrocytomas was higher than that of low-grade (grade 2) tumors (P = 0.0064). There were no significant differences in patient survival between the high- and low-LI groups of Bcl-2, Bcl-x, and Bax. Since the mechanism and regulation of apoptosis are still unclear, it seems difficult to use the Bcl-2 family as a biological marker in predicting the prognosis of patients with astrocytomas. PMID- 11908869 TI - Drug-resistance gene expression and progression of astrocytic tumors. AB - To clarify the influence of biochemotherapy on the progression of astrocytic tumors, the expression of O6-methylguanine DNA-methyltransferase (MGMT) mRNA, as well as of other drug-resistance- and drug-sensitivity-related genes such as multidrug resistance gene 1, multidrug resistance-associated protein, glutathione S-transferase-pi, DNA topoisomerase II, and interferon receptor mRNA, and the interferon regulatory factor (IRF)-1 and -2 ratios in gliomas were investigated by quantitative reverse transcription-polymerase chain reaction (RT-PCR). The mean MGMT/beta2-microglobulin (beta2-MG) ratio for 130 neuroepithelial tumors was 8.2 +/- 17.8. The mean ratio of 45 glioblastomas was significantly higher than that for the other 85 tumors. In contrast, the mean of 26 low-grade gliomas was significantly lower than that of other tumors. The mean IRF-1/IRF-2 ratio of 16 other brain tumors that mainly consisted of medulloblastomas was significantly greater than that of the other 114 tumors. Almost no significant differences were observed between primary and recurrent tumors in the expression of any gene, and before and after therapy with corresponding drugs. The mean MGMT/beta2-MG ratio in primary glioblastomas was significantly higher than that in secondary tumors. These findings suggest that native drug resistance is more important than acquired resistance when glioma therapy is considered. PMID- 11908870 TI - Primary malignant lymphoma of the brain: analysis of MMAC1 (PTEN) tumor suppressor gene. AB - With the use of RT-PCR (reverse transcriptase-polymerase chain reaction), Northern blot analysis, and Western blot analysis, seven primary brain lymphomas were examined for the state of the MMACI tumor suppressor gene. Nucleotide analysis of RT-PCR clones revealed no abnormality in the MMAC1 coding sequence in each case. Although Northern blot revealed variation among cases in the signal intensities for MMAC1 mRNA, Western blot revealed a distinct MMAC1 protein band in all cases, suggesting that the actual MMAC1 expressions were similar. In Western blot analysis of phosphorylated Akt (p-Akt), which is regulated positively by PI3K (phosphoinositide-3 kinase) and negatively by MMAC1, all the lymphomas revealed an Akt band but not a p-Akt band, suggesting that the MMAC1 phosphatase activity was maintained in each case. These findings suggest that the MMAC1 gene is normal in its coding sequence, gene expression, and phosphatase activity in the lymphomas. Thus, unlike the p16 and p15 tumor suppressor genes, which are frequently deleted and inactivated in brain lymphoma and represent a striking contrast to systemic lymphoma, MMAC1 may not play an important role in carcinogenesis in this tumor, as in the systemic counterpart. PMID- 11908871 TI - Localization of gelatinase activities in glioma tissues by film in situ zymography. AB - Malignant gliomas are characterized by their invasiveness and angiogenesis, which are mediated by metalloproteinases (MMPs). In this study, localization of gelatinase activities in six cases of glioblastoma, two cases of anaplastic astrocytoma, and six cases of low-grade astrocytomas was investigated by film in situ zymography (FIZ). FIZ is a novel technique, because it allows preservation of tissue structure with gelatinolytic activity and can detect net MMP activity in tumor tissues. In malignant gliomas, gelatinolytic activities were extensively localized in the vessel walls in the viable tumor areas and peritumoral areas as well as the tumor cell itself. These findings suggest that FIZ is a useful tool not only to detect tumor net MMP activities but also to detect angiogenic features in the invading front of malignant gliomas. PMID- 11908872 TI - Prognostic value of Ki-67 (MIB-1) and p53 in ependymomas. AB - To determine whether Ki-67 (MIB-1) and p53 have prognostic value in ependymomas, clinicopathologic study was undertaken in 29 patients with this tumor. The clinical course correlated well with the histological grade according to the World Health Organization (WHO) grading system, and it was the worst in patients with anaplastic ependymoma. The percent expression of MIB-1 and p53 correlated with the histological grade of malignancy. With regard to the subtypes of benign ependymoma, the clinical course was the worst in clear-cell ependymoma, which had a significantly higher expression of MIB-1 and p53 than the other subtypes. Tanycytic ependymoma showed the most benign clinical course and the lowest expression of MIB-1 and p53. Although the WHO grading generally tended to correlate with the clinical course of ependymomas, these two subtypes--clear-cell ependymoma and tanycytic ependymoma--exhibited biological properties different from those of other grade II ependymomas. PMID- 11908873 TI - Anaplastic meningioma with papillary, rhabdoid, and epithelial features: a case report. AB - A 74-year-old man manifested disturbed consciousness and right hemiparesis. Computed tomography revealed a left frontal parasagittal meningeal tumor with extensive peritumoral brain edema and skull invasion. Subtotal removal was performed. Five years later, he underwent two more operations of massive recurrences. Pathological studies revealed anaplastic meningioma with two different histological areas. One was an epithelial and meningothelial area, and the other was a papillary and rhabdoid area. In the papillary and rhabdoid area, small tumor cells with a high nucleus/cytoplasm ratio proliferated densely around the dilated central capillaries with a pseudopapillary pattern. Many rhabdoid cells (vimentin ++, cytokeratin AE1/AE3 +, epithelial membrane antigen [EMA] + +) tended to be distributed far from the central capillaries. There were many mitotic figures near the central vessels. Dense MIB1-positive nuclei were also observed near the central vessels. The trabecular pattern of the tumor cells in the epithelial area was quite different from the histological features of chordoid meningioma. PMID- 11908874 TI - Detection of Epstein-Barr virus DNA and expression of CD30 antigen in primary anaplastic diffuse large B-cell lymphoma of the brain. AB - We describe a case of primary anaplastic diffuse large-cell lymphoma arising in the central nervous system (CNS). Primary CD30-positive anaplastic diffuse large B-cell lymphoma of the brain is very rarely reported. Given that this tumor is immunohistochemically heterogeneous, polymerase chain reaction (PCR) and Epstein Barr virus (EBV) analysis of tumor DNA are essential techniques for early and accurate histological diagnosis in these CD30-positive cerebral lymphoma cases. We report an early CD30- and EBV-positive anaplastic diffuse large B-cell lymphoma in the CNS that was diagnosed not only from the immunohistochemical study and MRI findings, but also from the genotype confirmations. This tumor was documented to have EBV episomes of monoclonal origin by PCR analysis of immunoglobulin gene rearrangement. PMID- 11908875 TI - Different responses of benign and atypical meningiomas to gamma-knife radiosurgery: report of two cases with immunohistochemical analysis. AB - Recent reports have shown that gamma-knife radiosurgery provides a safe and effective strategy for the management of brain tumors. To evaluate the role of stereotactic radiosurgery in the management of meningiomas, we investigated the histopathology of two patients. The patients, a 37-year-old man and a 54-year-old woman, presented with visual field disturbance or headache. Imaging studies demonstrated intracranial meningiomas--tentorial and sphenoid ridge, respectively. Each patient undewent subtotal surgical resection (more than 90% in both patients), followed by gamma-knife radiosurgery of the remnant tumor marginal doses of 15 Gy. Pathological examination of the original tumors revealed a meningothelial meningioma and an atypical meningioma, respectively. Enlargement of the remnant tumors 4 months after radiosurgery resulted in total surgical resection in both patients. Thirteen months later, the patient with the atypical meningioma underwent a third operation for early recurrence of the tumor. Histopathology was investigated, and MIB-1, p53, and bcl-2 labeling indexes (LI) were analyzed immunohistochemically. Histopathologically, the specimens showed necrosis and intratumoral vessel obliteration after radiosurgery in both cases. However, more remnant tumor cells survived in the atypical meningioma. Immunohistochemically, increased wild-type p53, decreased bcl-2 expression, and decreased MIB-1 LI were observed in the benign meningioma. In the atypical meningioma, on the contrary, MIB-1 LI was decreased and mutant-type p53 and bcl-2 expression were unchanged. The specimen from the third operation revealed an anaplastic meningioma, and MIB-1 LI was markedly increased. These findings suggest that the efficacy of radiosurgery may differ between benign and atypical meningiomas. PMID- 11908876 TI - Expression of vascular endothelial growth factor (VEGF) and its two receptors in diffusely infiltrating astrocytomas and relationship to proliferative activity of tumor cells. AB - We studied the relationships among vascular endothelial growth factor (VEGF), its receptors (Flt1 and Flk1), and MIB-1. Their expression in 47 diffusely infiltrating astrocytomas obtained at surgery or autopsy was investigated by the ABC method and analyzed quantitatively. The positive rate of VEGF in tumor cells was higher than that in endothelial cells, and Flk1 was lower in tumor cells (P < 0.01, 0.01), whereas Flt1 in both tumor cells and endothelial cells was found at similar levels (P > 0.05). In tumor cells, VEGF became high with increased histological grades (P < 0.01). whereas both Flt1 and Flk1 were higher in grade 4 than in grades 2 and 3 (P < 0.01, 0.05). VEGF, Flt1, and Flk1 in endothelial cells were also highly expressed in grade 4 (P < 0.01). The distribution of MIB-1 positive nuclei in grade 4 was similar to VEGF, and the percent of positivity from grade 2 to grade 4 also increased (P < 0.01). There was a linear positive correlation between VEGF and both Flt1 and Flk1 in both tumor cells and endothelial cells (P < 0.01). So was the percent of positivity with VEGF, Flt1, and Flk1 in tumor cells and endothelial cells (P < 0.01). The experiment suggests that VEGF may act as a growth factor for both endothelial cells and tumor cells. VEGF, Flt1, and Flk1 can be considered as indicators of the malignancy potential of diffusely infiltrating astrocytomas. The expression of VEGF and the two receptors may be affected by the proliferative activity of tumor cells. PMID- 11908877 TI - Phenotypic changes associated with exogenous expression of p16INK4a in human glioma cells. AB - The tumor suppressor p16/CDKN2A/INK4a gene is frequently mutated, mostly by homozygous deletions in high-grade gliomas. Although the p16 protein suppresses cell proliferation primarily through inhibition of cell-cycle progression at the G1 phase, other phenotypic changes in glioma cells associated with p16INK4a alterations have not been fully described. To determine the roles of p16 alterations in glioma formation, we have established ecdysone-driven inducible p16 expression in the human glioblastoma cell line CL-4, which were derived from p16-null U87MG cells. Here we show that exogenous p16 expression in CL-4 cells results in morphological changes, with large and flattened cytoplasm, which are associated with increased formation of cytoplasmic actin-stress fibers and vinculin accumulation in the focal adhesion contacts. Adhesion of CL-4 cells to extracellular matrix proteins, such as laminin, fibronectin, and type IV collagen, significantly increased upon exogenous p16 expression, which correlated with increased expression of integrin alpha5 and alphav. Expression of a small GTP-binding protein, Rac, also decreased. Following epidermal growth factor stimulation, phosphorylation of MAP kinases ERK1 and 2 and induction of an early immediate gene product, c-Fos, were significantly reduced in CL-4 cells with p16 expression. These results suggest that the tumor suppressor p16 may exert its antitumor effects through modulation of multiple aspects of glioblastoma phenotypes, including proliferation, invasiveness, and responsiveness to extracellular growth stimuli. PMID- 11908878 TI - The antimalarials quinacrine and chloroquine potentiate the transplacental carcinogenic effect of ethylnitrosourea on ependymal cells. AB - Quinacrine and chloroquine, two widely used antimalarials, bind strongly to deoxyribonucleic acid, thus preventing mutagenesis. We studied a possible chemoprotective effect of these substances on carcinogenesis of the nervous system induced in Wistar rats by transplacental administration of ethylnitrosourea. One experimental group consisted of rats born from mothers treated with quinacrine prior to prenatal exposure to ethylnitrosourea; a second group consisted of rats chronically treated with chloroquine after prenatal exposure to ethylnitrosourea. When compared with controls, no significant differences were observed in tumor incidence. However, early tumor growth was observed in both rats treated with quinacrine (P < 0.0004) and rats treated with chloroquine (P < 0.02). These differences were due mostly to rapid development of ependymomas of the spinal cord. Our results suggest that quinacrine and chloroquine do not prevent the structural alterations induced in DNA by ethylnitrosourea, which lead, in the long term, to a high incidence of neoplasms in the nervous system. Moreover, the antimalarials studied seem to promote the carcinogenic effects of ethylnitrosourea on ependymal cells. PMID- 11908880 TI - Physiological implications of mechanical effects of +Gz accelerations on brain structures. AB - BACKGROUND: In certain flight configurations, fighter pilots are exposed to high Gz acceleration (G) which may induce inflight loss of consciousness (G-LOC). When acceleration is of high amplitude, and the onset rate very rapid, G-LOC can occur extremely suddenly. HYPOTHESIS: Mechanisms other than brain hypoxia could be involved, enhancing its effects. In order to study the mechanical effects induced by such accelerations on cerebral structures, we estimated the stresses imposed on cerebral tissue when the brain is exposed to +Gz acceleration forces. METHODS: An "in vitro" experiment was performed to measure brain deformations during centrifugation. A finite element model of the brain was formulated. RESULTS: Computations indicate that traction and shear stresses are enhanced around the tentorial notch, and that compression stresses increase at the base of the cerebral hemispheres. CONCLUSION: The amplitude of these stresses is not sufficient to directly disturb proper nerve cell functioning. However, they could interfere with brain vessels as external surface forces, thus enhancing vessel collapse and brain ischemia. PMID- 11908879 TI - Novel approach to analysis of in vitro tumor angiogenesis with a variable pressure scanning electron microscope: suppression by matrix metalloproteinase inhibitor SI-27. AB - Degradation of basement membrane by metalloproteinases (MMP) is a critical step in tumor angiogenesis. To evaluate in vitro angiogenesis, several models have been employed, including bovine cornea, fenestrated rat brain, Matrigel, and others. These models did not provide quantitative analysis of capillary formation. The current study aimed for a novel approach to in vitro assay of angiogenesis with a "wet scanning electron microscope (SEM)" to investigate suppression of tumor angiogenesis by the MMP inhibitor, SI-27. The effects of noncytotoxic concentrations of SI-27 (1-100 microM) were determined on nonmitogenic vascular endothelial growth factor (VEGF) (10 ng/ml)-mediated cell motility and in vitro angiogenesis of human umbilical vein endothelial cells (HUVECs). Activities of MMP and tissue inhibitor of metalloproteinase (TIMP) were determined by enzyme-linked immunosorbent assay (ELISA). Subsequently, the inhibitory effect of SI-27 was examined on in vitro angiogenesis stimulated by supernatants of human glioma cell lines (U87MG, U251MG, or U373MG). In vitro angiogenesis was quantitatively analyzed with a variable-pressure SEM. Cell motility and in vitro angiogenesis by HUVECs were significantly increased by VEGF along with elevated MMP-1 and -2 activity, whereas SI-27 significantly suppressed VEGF-mediated in vitro angiogenesis and inactivated both MMP-1 and MMP-2, but not inhibited cell motility. The angiogenesis promoted by glioma supernatants showed a significant reduction in the presence of SI-27. SI-27, a novel MMP inhibitor, inhibited tumor angiogenesis in vitro. It can be anticipated to prevent tumor growth through its angiosuppressive effect. Quantitative analysis with a variable pressure SEM is a novel approach to in vitro angiogenesis assay. PMID- 11908881 TI - Electromagnetic interference in pacemakers in single-engine fixed-wing aircraft: a European perspective. AB - BACKGROUND: The aim of this study is to evaluate the possible interference by avionics with cardiac pacemakers of aircrew or passengers/patients in single engine fixed-wing aircraft. Pacemakers which are implanted in patients are in part electromagnetic sensors and can be subject to interference from various external electromagnetic sources. Although modern (chip-based) pacemakers are effectively shielded from electromagnetic interference (EMI), the magnitude of electromagnetic radiation in cockpits of general aviation aircraft is higher and of a different nature than experienced in daily life. An increasing number of pacemaker-bearing individuals are being transported by air. However, the possible EMI with modern types of pacemakers during flight has not been investigated until now. METHOD: In order to evaluate the effect of EMI on five modern types of pacemakers in the cockpit environment of a single-engine fixed-wing aircraft, we have subjected the pacemakers, each implanted into an artificial thorax, to a series of test flights. Each pacemaker was equipped with data logging capabilities which were used for detection of possible EMI. After each flight, the pacemakers were examined by means of the dedicated programmers. In addition, two single lead ventricular pacemakers (VVI) were analyzed by means of beat-to beat Holter recordings during two separate flights. This enabled an exact analysis of pacemaker function and of possible EMI. RESULTS: No effect of EMI could be detected in any of the pacemakers by interrogating their internal counters after the test flights. In addition, no signs of EMI could be detected on the Holter recordings of the VVI pacemakers. CONCLUSION: We conclude that modern pacemakers are unaffected by EMI in the cockpit environment of single engine fixed-wing aircraft. PMID- 11908882 TI - Postural control and venous gas bubble formation during hypobaric exposure. AB - BACKGROUND: Earlier studies have shown that acute hypoxia at simulated altitudes up to 18,000 ft affects postural control. The main objective of this study was to investigate whether this is caused by hypoxia or by other effects of reduced barometric pressure. Doppler monitoring was included to rule out venous gas emboli (VGE) as a possible cause of disturbed postural control. A secondary objective was to evaluate two conventional altitude chamber training profiles regarding release of VGE. HYPOTHESIS: Chamber flights up to 18,000 ft affect postural control due to acute hypoxia or other effects of reduced barometric pressure such as bubble formation. VGE probably will not be formed at the altitude chamber flight profiles and procedures selected for this study. METHODS: Repeated registrations of postural control and Doppler monitoring for detection of possible VGE were performed on 12 subjects before, during, and after exposure to two different altitude chamber flight profiles. In chamber flight profile 1 the subjects were first preoxygenated for 45 min and then exposed to a normoxic environment at altitudes of 25,000, 18,000, 14,000, and 8000 ft. Chamber flight profile 2 consisted of an 80 min exposure to 14,000 ft without preoxygenation or supplemental oxygen for the first 60 min. RESULTS: In chamber flight profile 1, where normoxic conditions were achieved during all balance testing, no significant changes in postural control were found. No VGE were observed and no subjective dizziness was reported during this exposure. In chamber flight profile 2, a significant influence on postural control was reported for the eyes-open condition, when breathing air at 14,000 ft. These changes normalized when reaching ground level. VGE were observed in one of the 12 subjects after 75 min at 14,000 ft. Another subject complained of severe dizziness during the initial part of the decompression to 14,000 ft, and was excluded from further experiments. CONCLUSIONS: Changes in postural control at altitudes up to 18,000 ft is probably due to acute hypoxia. VGE may form during acute altitude exposure to 14,000 ft. PMID- 11908883 TI - Perception of the cabin attitude changes in hypergravity. AB - BACKGROUND: The G-excess illusion is becoming increasingly recognized as a cause of aviation fatalities. Studies of this illusion have looked at perception of subjects' orientation by moving the head during hypergravity, but independent of the pilot's head movement with respect to aircraft. This illusion can also occur by aircraft motion, but this has not been studied extensively. OBJECTIVE: The aim of this study was to investigate the subject's perception of orientation to the simulator cab attitude changes at 1.6 G without making any head movement with respect to the cab, and assess the feasibility of simulating the G-excess illusion on the ground with a centrifuge-like spatial disorientation simulator. METHODS: The 1.6-G force field was provided by the gravitoinertial force (GIF) of the simulator when it made an off-center (planetary) rotation at a constant velocity of 130 degrees x s(-1). Eleven subjects' perceptions of orientation of the cab attitudes were collected respectively by their report before and after certain cab tilt, in a roll plane of 1.6 G. RESULTS: When the cab was tilted 20 degrees at 1.6 G, the subjects perceived the angle to be 48.6 +/- 39.4 degrees. CONCLUSION: Most subjects experienced an exaggerated sensation to the cab attitude changes in roll plane. G-excess illusion can be generated in a centrifuge-like device on the ground. PMID- 11908884 TI - The age 60 rule: age discrimination in commercial aviation. AB - BACKGROUND: The Federal Aviation Administration's Age 60 Rule, promulgated in 1959, prohibits airline pilots from working in Part 121 operations once they have reached the age of 60. The Age 60 Rule remains a most contentious and politically sensitive topic, with challenges to the Rule currently mounted in both legislative and legal arenas. METHODS: An extensive review of the medical literature was accomplished using MEDLINE. Pertinent Federal Regulations were examined. Legal proceedings and public domain documents were noted. Letters and personal communication were solicited where necessary information could not be ascertained by other means. RESULTS: The Age 60 Rule was not based on any scientific data showing that airline pilots aged 60 and older were any less safe than younger pilots, and there is evidence to indicate that the choice of age 60 was actually based on economic rather than safety considerations. Airline pilots consistently exceed general population norms for longevity, physical health, and mental abilities. Fear of an adverse pilot health event causing a crash in standard multi-crew operations is not justified. For decades, airline pilots under age 60 have been granted the means to demonstrate their fitness for flying by taking medical, cognitive, and performance evaluations that are denied to airline pilots when they reach age 60. Actual flight experience demonstrates that older pilots are as safe as younger pilots. International aviation experience indicates that abolishing the Age 60 Rule will not compromise aviation safety. CONCLUSION: There appears to be no medical, scientific, or safety justification for the Age 60 Rule. As such, perpetuation of the Age 60 Rule, where age alone is used as the single criterion of older pilot fitness, represents age discrimination in commercial aviation. PMID- 11908885 TI - Outbreak of head pediculosis during SFINCSS-99 isolation experiment. AB - This short case report presents the development and treatment of a head lice outbreak during a space simulation experiment. The confinement experiment was conducted from July 1999 to March 2000 at the State Research Center of the Russian Federation Institute for Biomedical Problems in Moscow. Following a symptom period of up to 9 wk before diagnosis and treatment, 3 wk of treatment were required to remove all nits from those infected. Recommendations are made with regard to prevention of such infections during space missions. Treatment altered performance and comfort of crews, took time out of other duties, and even affected relations between crewmembers. The case is made for the development of a public health agenda in space research and medical space programs. PMID- 11908886 TI - Influence of microgravity on plasma levels of gastroenteropancreatic peptides: a case study. AB - BACKGROUND: No data are available about the short- or long-term influences of microgravity in space on the release of gastroenteropancreatic peptides, although these peptides are involved in the regulation of gastrointestinal functions. METHODS: Fasting plasma samples were gained during the EUROMIR-94 mission from a European Space Agency (ESA) astronaut who experienced no signs of space motion sickness in orbit. Plasma concentrations of nine gastroenteropancreatic peptides were measured with sensitive and specific radioimmunoassays. RESULTS: Fasting plasma levels of motilin, pancreatic polypeptide (PP), vasoactive intestinal peptide (VIP), and secretin were increased and plasma level of cholecystokinin (CCK) was decreased by acute exposure of the astronaut to microgravity. Chronic (4 wk) exposure caused an enhancement of plasma CCK, motilin, neurotensin, VIP, and insulin whereas plasma concentrations of PP, secretin, gastrin, and somatostatin showed no changes. During the 25-d stay on MIR station plasma levels of CCK, motilin, and neurotensin increased. Short-time body rotations caused an elevation of plasma levels of PP but decreased plasma motilin. CONCLUSIONS: As the influence of microgravity on the peptide levels was not uniform, an effect due to other factors (e.g., change in fluid balance or body weight) is unlikely. Moreover, adaptive changes of some peptides occurred during the stay in orbit. The release of PP and motilin seems to be very sensitive to rotation forces. These results have to be confirmed in more subjects in space to be able to link changes of gastroenteropancreatic peptide release to alterations of gastrointestinal functions. PMID- 11908887 TI - Cabin air quality: an overview. AB - In recent years, there have been increasing complaints from cockpit crew, cabin crew, and passengers that the cabin air quality of commercial aircraft is deficient. A myriad of complaints including headache, fatigue, fever, and respiratory difficulties among many others have been registered, particularly by flight attendants on long-haul routes. There is also much concern today regarding the transmission of contagious disease inflight, particularly tuberculosis. The unanswered question is whether these complaints are really due to poor cabin air quality or to other factors inherent intlight such as lowered barometric pressure, hypoxia, low humidity, circadian dysynchrony, work/rest cycles, vibration, etc. This paper will review some aspects relevant to cabin air quality such as volatile organic compounds (VOCs), carbon dioxide (CO2), carbon monoxide (CO), ozone (O3), particulates, and microorganisms, as well as the cabin ventilation system, to discern possible causes and effects of illness contracted inflight. The paper will conclude with recommendations on how the issue of cabin air quality may be resolved. PMID- 11908888 TI - The correlation between aerobic fitness and motion sickness susceptibility. AB - Susceptibility to motion sickness has been linked to aerobic fitness in several studies, however, these studies have not elucidated the underlying physiological mechanism by which increased aerobic fitness is related to a decreased ability to tolerate motion sickness stimuli. This pilot study provides further evidence of a relationship between aerobic fitness and motion sickness susceptibility. It also suggests that aerobic capacity is more specifically linked to signs and symptoms of vasomotor origin including stomach discomfort, nausea and/or vomiting, headache, and diaphoresis. By independently correlating vasomotor susceptibility and neurogenic susceptibility to maximum oxygen uptake, we find that vasomotor symptoms in particular are significantly increased in aerobically fit individuals. Larger studies should be conducted to confirm this relationship. PMID- 11908889 TI - You're the flight surgeon. Altitude chamber-related symptoms. PMID- 11908890 TI - National Academy of Sciences report on cabin air quality. PMID- 11908891 TI - Keeping you informed of the latest advances in science and technology. PMID- 11908892 TI - The phylogenetic position of the pelobiont Mastigamoeba balamuthi based on sequences of rDNA and translation elongation factors EF-1alpha and EF-2. AB - The taxonomic position and phylogenetic relationships of the Pelobionta, an amitochondriate amoeboflagellate group, are not yet completely settled. To provide more information, we obtained sequences for the large subunit rDNA gene, the gene for translation elongation factor 1alpha, and for a large part of the gene encoding translation elongation factor 2 from a representative of this group, Mastigamoeba balamuthi (formerly Phreatamoeba balamuthi). The gene for the large subunit rDNA was unusually large compared to those of other protists, a phenomenon that had previously been observed for the gene encoding the small subunit rDNA. Phylogenetic reconstruction using a maximum likelihood method was performed with these sequences, as well as the gene encoding the small subunit rDNA. When evaluated individually, the M. balamuthi genes for the small and large subunit rDNAs and elongation factor 1alpha had a most recent common ancestor with either the Mycetozoa (slime molds) or with Entamoeba histolytica. A clade formed by M. balamuthi, E. histolytica, and Mycetozoa was not rejected statistically for any of the sequences. A combined maximum likelihood analysis using 3,935 positions from all molecules suggested that these three taxonomic units form a robust clade. We were unable to resolve the closest group to this clade using the combined analysis. These findings support the notion, which had previously been proposed primarily on cytological evidence, that both M. balamuthi and E. histolytica are closely related to the Mycetozoa and that these three together represent a major eukaryotic lineage. PMID- 11908893 TI - Serological affinities of the oyster pathogen Perkinsus marinus (Apicomplexa) with some dinoflagellates (Dinophyceae). AB - The protozoan oyster pathogen Perkinsus marinus is classified in the phylum Apicomplexa, although molecular-genetic and ultrastructural evidence increasingly concur on its closer phylogenetic relationship with the dinoflagellates. To test for evidence of serological epitopes common to P. marinus and dinoflagellates, we probed 19 free-living and 8 parasitic dinoflagellate, or dinoflagellate-like, species for cross-reactivity with polyclonal antibodies to P. marinus. Three of 19 free-living dinoflagellates (16%), and 7 of 8 parasitic dinoflagellates (88%) were labeled by anti-P. marinus antibodies. In reciprocal immunoassays using polyclonal antibodies to the Hematodinium sp. dinoflagellate parasite of Norway lobsters, Nephrops norvegicus, P. marinus and the same 7 parasitic dinoflagellates labeled by anti-P. marinus antibodies, were again labeled. The dinoflagellate-like parasite of prawns Pandalus platyceros was not labeled by either antibody reagent. These reciprocal results confirm the presence of shared antibody-binding epitopes on cells of P. marinus and several dinoflagellates. The apparent widespread serological affinity between P. marinus and the parasitic dinoflagellates suggests a closer phylogenetic link to the syndinean dinoflagellate lineage. The consistent failure of the dinoflagellate-like prawn parasite to bind either antibody reagent shows that this parasite is serologically distinct from both P. marinus and Hematodinium-species parasitic dinoflagellates. PMID- 11908894 TI - Laboratory and field-based studies of abundances, small-scale patchiness, and diversity of gymnamoebae in soils of varying porosity and organic content: evidence of microbiocoenoses. AB - Soil samples (varying in granularity) from four natural sites were cultured in microcosms to determine small-scale patchiness in abundance and diversity of gymnamoebae. Eighty grams of the same thoroughly mixed soil, either moistened with distilled water (- nutrients) or supplemented with an equivalent vol. of organically enriched water (+ nutrients), were placed in covered glass jars and incubated for 14 d (25 degrees C). Abundances (number/gram soil) were assessed in each of 3 core samples (5-10 mm apart). Assay precision was estimated to be +/- 4%. Abundances were similar in the 3 closely-spaced samples, but occasional samples had higher abundances, probably representing localized enriched sites ("nutrient hot spots"). Diversity within the triplicate, closely spaced samples varied substantially. Mean abundance and diversity of amoebae were consistently higher in organically enriched soil and in soil of increasing granularity. Field samples collected directly from two of the sites showed similar patterns of abundance and diversity as found in the experimental studies, indicating substantial small-scale compartmentalization of soil protist communities. These data provide evidence of soil eukaryotic microbiocoenoses and indicate that soil microfauna may encounter wide variations in resources and prey communities as they migrate within small distances of several millimeters or less. PMID- 11908895 TI - Transplasma membrane electron transport in Leishmania donovani promastigotes. AB - Leishmania donovani promastigotes are capable of reducing certain electron acceptors with redox potential at pH 7 down to -125 mV; outside the plasma membrane promastigotes can reduce ferricyanide. Ferricyanide has been used as an artificial electron acceptor probe for studying the mechanism of transplasma membrane electron transport. Transmembrane ferricyanide reduction by L. donovani promastigotes was not inhibited by such mitochondrial inhibitors as antimycin A or cyanide, but it responded to inhibitors of glycolysis. Transmembrane ferricyanide reduction by Leishmania appears to involve a plasma membrane electron transport chain dissimilar to that of hepatocyte cells. As with other cells, transmembrane electron transport is associated with proton release, which may be involved in internal pH regulation. The Leishmania transmembrane redox system differs from that of mammalian cells in being 4-fold less sensitive to chloroquine and 12-fold more sensitive to niclosamide. Sensitivities to these drugs suggest that transplasma membrane electron transport and associated proton pumping may be targets for the drugs used against leishmaniasis. PMID- 11908896 TI - Re-examining alveolate evolution using multiple protein molecular phylogenies. AB - Alveolates are a diverse group of protists that includes three major lineages: ciliates, apicomplexa, and dinoflagellates. Among these three, it is thought that the apicomplexa and dinoflagellates are more closely related to one another than to ciliates. However, this conclusion is based almost entirely on results from ribosomal RNA phylogeny because very few morphological characters address this issue and scant molecular data are available from dinoflagellates. To better examine the relationships between the three major alveolate groups, we have sequenced six genes from the non-photosynthetic dinoflagellate, Crypthecodinium cohnii: actin, beta-tubulin, hsp70, BiP, hsp90, and mitochondrial hsp10. Beta tubulin, hsp70, BiP, and hsp90 were found to be useful for intra-alveolate phylogeny, and trees were inferred from these genes individually and in combination. Trees inferred from individual genes generally supported the apicomplexa-dinoflagellate grouping, as did a combined analysis of all four genes. However, it was also found that the outgroup had a significant effect on the topology within alveolates when using certain methods of phylogenetic reconstruction, and an alternative topology clustering dinoflagellates and ciliates could not be rejected by the combined data. Altogether, these results support the sisterhood of apicomplexa and dinoflagellates, but point out that the relationship is not as strong as is often assumed. PMID- 11908897 TI - Establishment of a spinated type of Diplodinium rangiferi by transfaunation of the rumen ciliates of Japanese sika deer (Cervus nippon centralis) to the rumen of two Japanese shorthorn calves (Bos taurus taurus). AB - One liter of rumen fluid containing 4.7 x 10(4) ciliates/ml, representing four genera including nine species of ciliates from a Japanese sika deer was inoculated into two unfaunated Japanese shorthorn calves. Two weeks after inoculation, all species originally present in the inoculum were subsequently detected in the rumen fluid of one or both calves. Ciliate densities ranged from 10(5)-10(6) cells/ml over the remainder of the 33-wk experiment. The inoculum contained Diplodinium rangiferi. which lacks caudal appendages, as is characteristic for the species. However, three weeks later, the rumen fluid of both calves contained D. rangiferi, which possesses caudal appendages varying from a single spine to multiple spines with a complicated furcate appearance. The caudal spines of D. rangiferi did not disappear during the experiment, even when the diet of the calves was switched to the ration of sika deer from which the inoculum was obtained. PMID- 11908898 TI - Phylogenetic position of Blastocystis hominis and of stramenopiles inferred from multiple molecular sequence data. AB - Blastocystis hominis, a parasite of the human intestine, has recently been positioned within stramenopiles by the small subunit rRNA phylogeny. To further confirm its phylogenetic position using multiple molecular sequence data, we determined the nucleotide sequences putatively encoding small subunit ribosomal RNA, cytosolic-type 70-kDa heat shock protein, translation elongation factor 2, and the non-catalytic 'B' subunit of vacuolar ATPase of B. hominis (HE87-1 strain). Moreover, we determined the translation elongation factor 2 sequence of an apicomplexan parasite, Plasmodium falciparum, that belongs to alveolates. The maximum likelihood analyses of small subunit rRNA and cytosolic-type 70-kDa heat shock protein clearly demonstrated that B. hominis (HE87-1 strain) is positioned within stramenopiles, being congruent with the previous small subunit rRNA analysis, including the sequences of B. hominis (Nand strain) and a Blastocystis isolate from guinea pig. Although no clear resolution among major eukaryotic groups was obtained by the individual phylogenies based on the four molecules analyzed here, a combined analysis of various molecules, including these, clearly indicated that Blastocystis/stramenopiles are the closest relatives of alveolates. PMID- 11908899 TI - Not-adaptive behavior of isotropically heated, inert populations of Oxytricha bifaria (Ciliophora, Stichotrichia). AB - The physiological effects on isotropically heated populations of Oxytricha bifaria cultured at 24 degrees C were investigated. At 34.6 degrees C ciliates became inert, and did not adaptively react to either cold or warm microgradients; they neither moved towards the favorable cold thermal source nor escaped from the unfavorable warm one. The inert oxytrichas were only able to perform the Side Stepping Reaction (SSR) on the same spot. However, mobile ciliates at 31.6 degrees C reacted to the cold microgradient by immediately orienting themselves towards its source, without accelerating but reducing their SSR frequency. Moreover, in a warm microgradient such ciliates immediately increased their SSR frequency, then moved away from the thermal source. At 34.6 degrees C the behavior of ciliates was not-adaptive--not acting to guide the organisms to more favorable conditions--whereas at 31.6 degrees C it was still clearly adaptive. Therefore, the locomotory inertness of the oxytrichas at 34.6 degrees C was the result of thermal stress rather than their behavioral response to the environmental isotropy, in contrast to populations of the same species made inert at 9 degrees C. PMID- 11908900 TI - Ecology of the red-tide dinoflagellate Ceratium furca: distribution, mixotrophy, and grazing impact on ciliate populations of Chesapeake Bay. AB - Ceratium furca is a primarily photosynthetic dinoflagellate also capable of ingesting other protists. During 1995 and 1996, we documented the abundance of C. furca in Chesapeake Bay and determined grazing rates on prey labeled with fluorescent microspheres. Abundance usually remained below 20 cells ml(-1), although the species was capable of localized late-summer blooms (< or = 478 cells ml(-1)) in the more saline lower to mid-Bay region. Feeding rates ranged from 0 to 0.11 prey dinoflagellate(-1) h(-1) or from 0 to 37 pg C dinoflagellate( 1) h(-1) and were highest at lower salinities. Clearance rates averaged 2.5 +/- 0.35 microl dinoflagellate(-1) h(-1). Impact of C. furca feeding on prey populations was higher in the lower Bay, averaging 67% of Strobilidium spp. removed d(-1). Ingestion rates were positively correlated with prey abundance and dissolved inorganic nitrogen, but negatively with salinity, depth, dissolved inorganic phosphorus, and inorganic P:N ratio. Daily consumption of prey biomass by C. furca averaged 4.6% of body carbon, 6.5% of body nitrogen, and 4.0% of body phosphorus. with maximal values of 36, 51, and 32%, respectively. Thus, the ability to exploit an organic nutrient source when inorganic nutrients are limiting may give C. furca a competitive advantage over purely photosynthetic species. PMID- 11908901 TI - Microtubules mediate germ-nuclear behavior after meiosis in conjugation of Paramecium caudatum. AB - Microtubule dynamics in Paramecium caudatum were investigated with an anti-alpha tubulin antibody and a microinjection technique to determine the function of microtubules on micronuclear behavior during conjugation. After meiosis, all four haploid micronuclei were connected by microtubular filaments to the paroral region and moved close to this region. This nuclear movement was micronucleus specific, because some small macronuclear fragments transplanted from exconjugants never moved to the region. Only one of the four germ nuclei moved into the paroral cone and was covered by microtubule assembly (the so-called first assembly of microtubules, AM-I). This nucleus survived there, while the other three not in this region degenerated. The movement of germ nucleus was inhibited by the injection of the anti-alpha-tubulin antibody. The surviving germ nucleus divided once and produced a migratory pronucleus and a stationary pronucleus. Prior to the reciprocal exchange of the migratory nuclei, microtubules assembled around the migratory pronuclei again (the so-called second assembly of microtubules, AM-II). Then, the migratory pronucleus moved into the partner cell and fused with the stationary pronucleus. Thus, microtubules appear to be indispensable for nuclear behavior: they enable migration of postmeiotic nuclei to the paroral region and they permit the survival of the nucleus at the paroral cone. PMID- 11908902 TI - Arginine kinase: a common feature for management of energy reserves in African and American flagellated trypanosomatids. AB - This work reports the characterization of an arginine kinase in the unicellular parasitic flagellate Trypanosoma brucei, the etiological agent of human sleeping sickness and Nagana in livestock. The arginine kinase activity, detected in the soluble fraction obtained from procyclic forms, had a specific activity similar to that observed in Trypanosoma cruzi, about 0.2 micromol min(-1) mg(-1). Western blot analysis of T. brucei extracts revealed two bands of 40 and 45 kDa. The putative gene sequence of this enzyme had an open reading frame for a 356-amino acid polypeptide, one less than the equivalent enzyme of T. cruzi. The deduced amino acid sequence has an 82% identity with the arginine kinase of T. cruzi, and highest amino acid identities of both trypanosomatids sequences, about 70%, were with arginine kinases from the phylum Arthropoda. In addition, the amino acid sequence possesses the five arginine residues critical for interaction with ATP as well as two glutamic acids and one cysteine required for arginine binding. The finding in trypanosomatids of a new phosphagen biosynthetic pathway, which is not present in mammalian host tissues, suggests this enzyme as a possible target for chemotherapy. PMID- 11908903 TI - A structurally deviant member of the Euplotes raikovi pheromone family: Er-23. AB - Pheromones of Euplotes raikovi form a homologous family of proteins with 37- to 40-amino acid residues, including six cysteines that form three strictly conserved disulfide bridges. The determination of the primary structure of the pheromone Er-23, which was isolated from cells derived from natural populations of E. raikovi that secrete the other known pheromones, has now revealed a novel structure type. The polypeptide chain of this pheromone contains 51 residues, 10 of which are cysteines presumably involved in the formation of five disulfide bridges, and lacks a carboxyl-terminal tail following the last cysteine of the sequence. The elongation of the Er-23 molecule is presumed to result from multiple events of gene duplication starting from an ancestral motif Xxx(2-4)-Cys Xxx(5-7)-Cys. PMID- 11908904 TI - Dietary cis and trans monounsaturated and saturated FA and plasma lipids and lipoproteins in men. AB - Trans monounsaturated fatty acids (TFA) are hypercholesterolemic compared to oleic acid to a degree approaching or equivalent to saturated FA. However, it is unknown to what extent these effects may be due to cholesterol lowering by oleic acid rather than elevation by saturated FA and TFA. In order to better understand the impact of replacing TFA in foods, it is first necessary to know the relative lipid-modifying effects of the major FA that change as TFA are lowered or removed. For 5 wk, 50 normocholesterolemic men were fed controlled diets providing approximately 15% of energy from protein, 39% from fat, and 46% from carbohydrate in a randomized, 6 x 6, crossover design. Eight percent of energy was replaced across diets with the following: carbohydrate (CHO) (1:1 simple to complex); oleic acid (OL); TFA; stearic acid (STE); TFA/STE (4% of energy from each); carbon 12:0-16:0 saturated FA (LMP). LDL cholesterol concentrations (mmol/L) were as follows (different superscripts indicate significance at P < or = 0.01): OL 2.95a; CHO 3.05a,b; STE 3.10b,c; LMP 3.21c,d; TFA + STE 3.32d,e; and TFA 3.36e. HDL cholesterol concentrations (mmol/L) were as allows: STE 1.16a; TFA 1.16a,b; TFA/STE 1.17a,b; CHO 1.19b; OL 1.24c; and LMP 1.30d. Triacylglycerides were highest after STE (1.13) and lowest after OL (0.88) (P < 0.001). Thus, compared to the carbohydrate control diet, TFA raised LDL cholesterol at least equivalent to LMP but had no effect on HDL cholesterol; STE had no effect on LDL cholesterol but lowered HDL cholesterol; LMP raised both LDL cholesterol and HDL cholesterol; and oleic acid raised HDL cholesterol but had no effect on LDL cholesterol. PMID- 11908905 TI - Maternal supplementation with CLA decreases milk fat in humans. AB - CLA refers to isomers of octadecadienoic acid with conjugated double bonds. The most abundant form of CLA (rumenic acid (RA): c9,t11-18:2) is found in milk and beef fat. Further, CLA supplements containing RA and t10,c12-18:2 are now available. Consumption of commercially produced CLA has been shown to decrease adipose accretion in growing laboratory and production animals and cause milk fat depression in cows. We tested the hypothesis that CLA supplementation would increase milk CLA concentration and decrease milk fat content in humans. Breastfeeding women (n = 9) participated in this double-blind, placebo controlled, crossover study divided into three periods: intervention I (5 d), washout (7 d), and intervention II (5 d). Women were randomized to treatment order. During each intervention period, women consumed 1.5 g of CLA supplement or placebo (olive oil) daily; during the washout period, no supplements were consumed. Milk was collected by complete breast expression on the final day of each period; milk output was estimated by 24-h weighing on the penultimate day of each intervention period. Milk RA and t10,c12-18:2 concentrations were greater (P < 0.05) during the CLA treatment period as compared to the placebo period. Milk fat content was significantly lower during the CLA treatment, as compared to the placebo treatment (P < 0.05). Data indicate no effect of treatment on milk output. Therefore, it would be prudent that lactating women not consume commercially available CLA supplements at this time. PMID- 11908906 TI - Characterization of the adipose tissue atrophy induced by peroxisome proliferators in mice. AB - In the present study, we characterized the effects of peroxisome proliferators (PP) on adipose tissue in mice. Treatment with potent PP, such as perfluorooctanoic acid (PFOA), 2-methyl-2-(p(1,2,3,4-tetrahydroxy-naphthyl) phenoxy)propionic acid, (4-chloro-6-(2,3-xylidino)2-pyrimidinylthio) acetic acid, and di(2-ethylhexyl)phthalate, caused dramatic decreases in adipose tissue weight, whereas the moderately potent PP, acetylsalicylic acid, had a relatively weak effect. This decrease in weight reflects a loss of fat from adipocytes rather than a loss of cells, as demonstrated by constant DNA content. The dose dependency and time-course experiments indicate that peroxisome proliferation occurs simultaneously with or prior to adipose tissue atrophy. Thus, hepatic peroxisome proliferation might result in the increased mobilization of lipids and lipid utilization in liver. The enhanced adipose tissue hormone-sensitive lipase (HSL) activity and down-regulated lipoprotein lipase (LPL) activity observed upon PP treatment might, at least in part, explain the loss of fat via increased FA release from adipocytes and/or decreased FA uptake from the circulation, respectively. In addition, the possible involvement of the increased tumor necrosis factor alpha expression found upon PFOA treatment in reducing the insulin sensitivity of adipose tissue and thereby altering LPL and HSL activities is discussed. PMID- 11908907 TI - Evidence that leptin contributes to intestinal cholesterol absorption in obese (ob/ob) mice and wild-type mice. AB - In the present study, the effect of leptin on intestinal cholesterol absorption was investigated in C57 BL/6 OlaHsd Lep(ob)/Lep(ob) obese (ob/ob) mice and lean C57 BL/6 (wild-type) mice. Animals were treated either with or without recombinant leptin for 2 wk. Cholesterol absorption was measured by the constant isotope feeding method and indirectly by the ratio of campesterol to cholesterol in serum. In ob/ob mice, cholesterol absorption was significantly higher compared to wild-type mice [83.4 +/- 2.3% (SD) vs. 77.6 +/- 1.5%, P < 0.01]. Treatment with leptin significantly reduced cholesterol absorption in both ob/ob and wild type mice by 8.5 (P < 0.001) and 5.2% (P < 0.05), respectively. Serum concentrations of campesterol and the ratio of campesterol to cholesterol in ob/ob mice were significantly higher compared to wild-type mice (2.2 +/- 0.3 mg/dL vs. 1.2 +/- 0.3 mg/dL, P< 0.001; and 36.8 +/- 2.8 microg/mg vs. 28.0 +/- 3.3 microg/mg, P < 0.001). After treatment of ob/ob mice with leptin, concentrations of campesterol and its ratio to cholesterol were significantly lower (2.2 +/- 0.3 mg/dL vs. 1.0 +/- 0.2 microg/mg, P < 0.001; and 36.8 +/- 2.8 microg/mg vs. 13.2 +/- 2.2 microg/mg, P < 0.001, respectively). In wild-type mice, the ratio of campesterol to cholesterol in serum was also significantly lower after treatment with leptin (28.0 +/- 3.3 microg/mg vs. 22.6 +/- 5.0 microg/mg, P < 0.05). A significant positive correlation (r = 0.701, P < 0.01) between cholesterol absorption and the ratio of campesterol to cholesterol in serum was found. It is concluded that leptin contributes to intestinal cholesterol absorption in ob/ob mice and lean wild-type mice. PMID- 11908908 TI - Acute pancreatitis decreases pancreas phospholipid levels and increases susceptibility to lipid peroxidation in rat pancreas. AB - The objective of this study was to analyze whether acute pancreatitis leads to changes in the lipid composition and susceptibility to lipid peroxidation of pancreatic membranes. Total lipids, cholesterol, phospholipids, FA, and lipid peroxidation were determined in the pancreatic tissue of rats treated with cerulein and of control rats. In pancreatitic rats, significant decreases in membrane total phospholipid contents (P < 0.05) and in choline and ethanolamine glycerophospholipid levels (P < 0.05 and P < 0.01, respectively), with concomitant significantly higher values of their lysoderivative forms, were found. The cholesterol/phospholipid molar ratio increased by 26%. The unsaturation index of the FA profile decreased significantly (P < 0.01) as a consequence of a decrease in the arachidonic acid content. Incubation of membranes with xanthine oxidase/hypoxanthine-Fe2+/ADP resulted in an increase in the production of TBARS in pancreatitic rats compared to controls. In summary, acute pancreatitis causes changes in the lipid composition of rat pancreatic crude membranes and a greater susceptibility of these membranes to lipid peroxidation. PMID- 11908909 TI - Gamma-tocopheryl quinone stimulates apoptosis in drug-sensitive and multidrug resistant cancer cells. AB - Chemotherapy-induced cell death is linked to apoptosis, and there is increasing evidence that multidrug-resistance in cancer cells may be the result of a decrease in the ability of a cell to initiate apoptosis in response to cytotoxic agents. In previous studies, we synthesized two classes of electrophilic tocopheryl quinones (TQ), nonarylating alpha-TQ and arylating gamma- and delta TQ, and found that gamma- and delta-TQ, but not alpha-TQ, were highly cytotoxic in human acute lymphoblastic leukemia cells (CEM) and multidrug-resistant (MDR) CEM/VLB100. We have now extended these studies on tumor biology with CEM, HL60 and MDR HL60/MX2 human promyelocytic leukemia, U937 human monocytic leukemia, and ZR-75-1 breast adenocarcinoma cells. gamma-TQ, but not alpha-TQ or tocopherols, showed concentration and incubation time-dependent effects on loss of plasma membrane integrity, diminished viable cell number, and stimulation of apoptosis. Its cytotoxicity exceeded that of doxorubicin in HL60/MX2 cells, which express MRP, an MDR-associated protein. Apoptosis was confirmed by TEM, TUNEL, and DNA gel electrophoresis. Kinetic studies showed that an induction period was required to initiate an irreversible multiphase process. Gamma-TQ released mitochondrial cytochrome c to the cytosol, induced the cleavage of poly(ADP-ribose)polymerase, and depleted intracellular glutathione. Unlike xenobiotic electrophiles, gamma-TQ is a highly cytotoxic arylating electrophile that stimulates apoptosis in several cancer cell lines including cells that express MDR through both P-glycoprotein and MRP-associated proteins. The biological properties of arylating TQ electrophiles are closely associated with cytotoxicity and may contribute to other biological effects of these highly active agents. PMID- 11908910 TI - Farnesol and geranylgeraniol: prevention and reversion of lovastatin-induced effects in NIH3T3 cells. AB - Mevalonic acid-derived intermediates in the cholesterol biosynthetic pathway have been recognized as being critical to the isoprenylation of a variety of growth regulating proteins, including those of the RAS superfamily. Treatment of cells with lovastatin, a hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, depletes cells of mevalonic acid and thus blocks the isoprenylation of proteins in the RAS superfamily. In NIH3T3 cells pretreated with lovastatin, subsequent addition of farnesol (FOH), but not geranylgeraniol (GGOH), reverses lovastatin's block of RAS isoprenylation. Neither FOH nor GGOH prevents lovastatin-induced inhibition of RAS isoprenylation when added to cells concurrently with lovastatin. In intact cells, 167 microM FOH and 125 microM GGOH decrease incorporation of [14C]acetate into cholesterol by approximately 50 and 75%, respectively. Results suggest that the radio-label from either [3H]FOH or [3H]GGOH is incorporated into cholesterol. Co-treatment of cells with lovastatin or mevalonic acid did not significantly alter [3H]FOH or [3H]GGOH incorporation into cholesterol. Lovastatin induces cell rounding; GGOH, but not FOH, both prevents and reverses lovastatin-induced cell rounding. These results provide additional evidence for the existence of a novel "isoprenoid shunt" that differentially utilizes FOH and GGOH as metabolic precursors for isoprenoids that have been depleted by lovastatin treatment. PMID- 11908911 TI - Dietary n-3 PUFA alter colonocyte mitochondrial membrane composition and function. AB - There is experimental evidence that dietary fish oil, which contains the n-3 fatty acid family, i.e., EPA and DHA, protects against colon tumor development, in part by increasing apoptosis. Since mitochondria can act as central executioners of apoptosis, we hypothesized that EPA and DHA incorporation into colonocyte mitochondrial membranes, owing to their high degree of unsaturation, would enhance susceptibility to damage by reactive oxygen species (ROS) generated via oxidative phosphorylation. This, in turn, would compromise mitochondrial function, thereby initiating apoptosis. To test this hypothesis, colonic crypts were isolated from rats fed either fish oil, purified n-3 fatty acid ethyl esters, or corn oil (control). Dietary lipid source had no effect on colonic mitochondrial phospholipid class mole percentages, although incorporation of EPA and DHA was associated with a reduction in n-6 fatty acids known to enhance colon tumor development, i.e., linoleic acid LNA) and its metabolic product, arachidonic acid (ARA). Select compositional changes in major phospholipid pools were correlated to alterations in mitochondrial function as assessed by confocal microscopy. The mol% sum of LNA plus ARA in cardiolipin was inversely correlated with ROS (P = 0.024). Ethanolamine glycerophospholipid ARA (P = 0.046) and choline glycerophospholipid LNA (P = 0.033) levels were positively correlated to mitochondrial membrane potential. In contrast, ethanolamine glycerophospholipid EPA (P = 0.042) and DHA (P = 0.024) levels were negatively correlated to mitochondrial membrane potential. Additionally, EPA and DHA levels in choline glycerophospholipids (P = 0.026) were positively correlated with caspase 3 activity. These data provide evidence in vivo indicating that dietary EPA and DHA induce compositional changes in colonic mitochondrial membrane phospholipids that facilitate apoptosis. PMID- 11908912 TI - Cyclodextrins: a potential tool for studying the role of glycerolipids in photosynthetic membranes. AB - A novel cyclodextrin derivative, i.e., permethylated-alpha-cyclodextrin (PM-alpha CD), was used for removing glycerolipids from spinach thylakoid membranes and investigating their role in photosynthetic activities. A three-step selective removal of each lipid class was observed in treated membranes. Up to a concentration of 4 mM, PM-alpha-CD (in the presence of 75 microg chlorophyll a+b/mL), PM-alpha-CD displayed a marked selectivity for anionic lipids [sulfoquinovosyldiacylglycerol (SQDG) and phosphatidylglycerol (PG)] in comparison with galactolipids. At this concentration, half of PG and SQDG were removed. Within this range of concentration, the volume response of treated thylakoids to variation of osmolarity, an indirect mean of verifying the structural integrity of the membrane, was not altered. Similarly, neither photosystem II (PSII) nor photosystem I (PSI) activity was affected. In contrast, the low-temperature fluorescence ratio F695/F740 drastically diminished from 1.45 to about 0.7, essentially due to the decrease of PSII fluorescence. The results derived from the fast fluorescence rise expressed in the form of a spider suggest that the fraction of inactivated (non-Q(A) reducing) reaction centers (RC) increases while the active (Q(A) reducing) RC remained intact. Raising the concentration of PM-alpha-CD from 4 to 7 mM resulted in a progressive but greater diminution of the galactolipid level than that of SQDG and PG. Within this concentration range, the integrity of the membrane was not altered, nor was either PSII or PSI activity, whereas the F695/F740 ratio decreased to about 0.45 as well as the fraction of inactivated RC. At concentrations above 7 mM of PM alpha-CD, the integrity of the membrane was impaired, resulting in a decrease of both electron transport activities. At all concentrations, PM-alpha-CD did not show any selectivity toward either the acyl chains of the lipid molecules or the molecular species of PG. The results are discussed in terms of the role of glycerolipids in thylakoid membrane function and the relationship of the chemical structure of PM-alpha-CD and its lipid removal capacity. PMID- 11908913 TI - Biosynthesis of arachidonic acid in the oleaginous microalga Parietochloris incisa (Chlorophyceae): radiolabeling studies. AB - The fresh-water green alga Parietochloris incisa is the richest plant source of the PUFA arachidonic acid (20:4n-6, AA). To elucidate the biosynthesis of AA in this alga we labeled cultures of P. incisa with radioactive precursors. Pulse chase labeling with acetate resulted in its incorporation via the de novo biosynthesis pathway of FA. However, labeled acetate was also utilized for the elongation of C16 and C18 PUFA. Labeling with [1-14C]oleic acid has shown that the first steps of the lipid-linked FA desaturations utilize cytoplasmic lipids. PC and diacylglyceryltrimethylhomoserine are the major lipids involved as acyl carriers for the delta12 and delta6 desaturations of oleic acid, leading sequentially to linoleic and gamma-linolenic acids. The latter is released from its lipid carrier and elongated to 20:3n-6, which is reincorporated primarily into PE and PC and finally desaturated to AA. Galactolipids, mostly monogalctosyldiacyl-glycerol (MGDG), serve as substrates for the chloroplastic delta12 desaturase and, apparently, the omega3 desaturation, common to higher plants and many green algae. The predominant sequence desaturates the 18:1/16:0 molecular species of MGDG stepwise to the 18:3n-3/16:3n-3 molecular species similar to the prokaryotic pathway of higher plants and green algae. PMID- 11908914 TI - Detection of the 15-acetate of prostaglandin E2 methyl ester as a prominent component of the prostaglandins in the gorgonian coral Plexaura homomalla. AB - 15R-Prostaglandin E2 (PGE2) methyl ester 15-acetate (1) was isolated from the R variety of the Caribbean sea whip coral Plexaura homomalla collected in the Florida Keys. It was present in coral samples from separate collections in 2-10% of the abundance of the major prostaglandin component, PGA2 methyl ester 15 acetate. The structure of 1 was assigned based on one- and two-dimensional 1H NMR, HPLC, and LC-MS analyses. A sample of the S-variety of P. homomalla was found to contain a similar abundance of the corresponding 15S product, prostaglandin E2 methyl ester 15-acetate. The significance of PGE acetylation is discussed in relation to the proposed mechanism of PGA synthesis in the coral. PMID- 11908915 TI - Synthesis of (S)-alpha-amino oleic acid. AB - An efficient synthesis of (S)-alpha-amino oleic acid was developed. The fully protected FA derivative was obtained in four steps starting from methyl (2S)-2 [bis(tert-butoxycarbonyl)amino]-5-oxopentanoate. These steps are (i) olefination of the starting aldehyde with the appropriate phosphonate anion, (ii) hydrogenation of the double bonds, (iii) controlled reduction of omega-ethyl ester to an aldehyde in the presence of alpha-methyl ester, and (iv) a Wittig reaction of the latter aldehyde with the suitable ylide. Free alpha-amino oleic acid was prepared after deprotection of the amino group followed by saponification in a total yield of 24%. N-tert-Butoxycarbonyl-protected amino oleic acid and the corresponding amino alcohol were prepared in high yield. The structures of the products have been established by various spectroscopic techniques. PMID- 11908916 TI - The role of novel antipsychotics in bipolar disorders. AB - Patients with bipolar disorder frequently receive antipsychotic agents during both the acute and maintenance phases of treatment. Conventional antipsychotics are effective against mania, but they may induce depressive symptoms and expose patients with bipolar disorder to increased risks of tardive dyskinesia. Recent studies have shown risperidone to be effective for acute mania, both as monotherapy and in combination with mood stabilizers; this agent has also shown efficacy as add-on maintenance therapy in open-label studies as it exhibited both antimanic and antidepressant effects. Olanzapine, another novel antipsychotic, is also effective against both manic and depressive symptoms and in the maintenance treatment as indicated by an open-label study. Data on other novel agents are more limited. PMID- 11908917 TI - Psychotropic drugs and adverse events in the treatment of bipolar disorders revisited. AB - Psychopharmacology research aims to expand the therapeutic ratio between efficacy, on the one hand, and adverse events and safety, on the other. The novel antipsychotics are now the antipsychotics of choice in the treatment of bipolar disorders. They have the advantages of potential antidepressant properties and low risks of extrapyramidal side effects and, especially, of tardive dyskinesia. However, novel antipsychotics may also have varying propensities to cause side effects, such as somnolence, hyperprolactinemia, weight gain (sometimes significant), and possibly diabetes mellitus. The increasing use of these novel agents requires careful assessment and monitoring of emergent side effects and diligent consideration of associated medical complications. Two new anticonvulsants, lamotrigine and topiramate, have recently shown promise in the treatment of bipolar disorders. Most of their adverse effects can be avoided by slow titration toward the recommended doses. In contrast to carbamazepine and valproic acid, topiramate may be associated with weight loss. PMID- 11908918 TI - Predictors of treatment response in bipolar disorders: evidence from clinical and brain imaging studies. AB - The clinical features of bipolar disorders can be correlated with responses to medications. Patients who respond to lithium, for example, often present differently from those who respond to divalproex or carbamazepine, but the correlations are relatively modest. Brain-imaging tools, such as positron emission tomography (PET), single photon emission computed tomography (SPECT), and functional magnetic resonance imaging (fMRI), can relate brain function to clinical features and medication responses. For example, in depression, it appears that prefrontal cortical function is decreased while subcortical anterior paralimbic activity is increased. Preliminary evidence suggests that baseline metabolism increases and decreases in the left insula may be associated with carbamazepine and nimodipine responses, respectively, and that cerebral lithium concentrations may correlate with antimanic effects. Although it is not yet a clinical tool for bipolar disorders, brain imaging provides useful research data to understand the fundamental neurobiology of mood disorders and to more effectively target therapeutics. PMID- 11908919 TI - Bipolar disorders and the effectiveness of novel anticonvulsants. AB - The discovery that valproic acid is helpful in the management of patients with rapid-cycling bipolar disorder led to an explosion of research culminating in the third-generation anticonvulsants. Refractory depressive phases are frequent in bipolar disorders. No studies to date have shown that gabapentin is effective in bipolar mania or hypomania. Lamotrigine may have a role in treating bipolar depressive episodes, but it is not a particularly effective antimanic agent. Topiramate has shown encouraging results in both depressed and manic bipolar patients, and it may also promote weight loss. The new anticonvulsants are promising agents for the treatment of bipolar disorders, but they are heterogeneous with regard to their efficacy, target symptoms, and adverse event profiles. PMID- 11908920 TI - Role of central 5-HT(2) receptors in fluoxetine-induced decreases in T lymphocyte activity. AB - Previous studies have demonstrated that fluoxetine administration decreases mitogen-induced T lymphocyte proliferation. The present studies were carried out to determine which receptor subtype(s) was involved and whether these effects on lymphocyte responses were centrally or peripherally mediated. Two hours following administration of the 5-HT(1A) agonist 8-OH-DPAT (1 mg/kg), there was no change in lymphocyte proliferation responses, whereas the 5-HT(2) agonist DOI (2.5 mg/kg) significantly decreased (80%) proliferation. Similarly, pretreatment with the 5-HT(2) antagonists ritanserin (5 mg/kg, 30 min) or ketanserin (5 mg/kg, 1 h) was found to completely antagonize the effects of fluoxetine on lymphocyte proliferation. Consistent with central 5-HT(2) receptor involvement, microinjection of DOI (50 microg) resulted in a decrease in lymphocyte proliferation similar to that observed following systemic administration. Furthermore, central administration of ketanserin (20 microg) prevented the suppressive effects of systemic fluoxetine. Collectively, these results suggest that decreases in mitogen-induced lymphocyte proliferation following acute fluoxetine administration was due to indirect effects of fluoxetine following the activation of central 5-HT(2) receptors. PMID- 11908921 TI - The effects of stress on memory cytotoxic T lymphocyte-mediated protection against herpes simplex virus infection at mucosal sites. AB - Psychological stress has been shown to affect many components of the innate and adaptive immune responses to a variety of pathogens including herpes simplex virus (HSV). Mucosal tissues are clinically relevant sites of infection with HSV as well as with many other common pathogens. However, there is a scarcity of experimental evidence that stress affects mucosal immunity. We have taken advantage of a murine model of HSV-specific immune protection that is mediated by only memory cytotoxic T lymphocytes (CTLm) specific for a single CTL recognition epitope within glycoprotein B of HSV-1 (gB498-505). This CTLm population is elicited by vaccination with a recombinant vaccinia virus, which expresses this epitope in the absence of any other HSV-encoded antigens. We report here that stress reduces the ability of gB498-505-specific CTLm to protect against a lethal intranasal or intravaginal HSV infection. Also, stress decreases the ability of these CTLm to limit virus levels at the mucosal site of infection but does not have a significant effect on the levels of virus in the innervating sensory ganglia. Finally, stress decreases protection against HSV-mediated pathology of the vaginal epithelium. These studies are the first to examine the effects of stress on CTLm activation and function in vivo. PMID- 11908922 TI - Chemical sympathectomy has no effect on the severity of murine AIDS: murine AIDS alone depletes norepinephrine levels in infected spleen. AB - Numerous studies have shown that alterations in sympathetic nervous system (SNS) function produced by beta-adrenergic receptor blockade or chemical sympathectomy can produce changes in T and B lymphocyte function and both innate and acquired immune responses. However, fewer studies have investigated changes in immune response following SNS alterations in animal models of disease. We tested whether blocking SNS activity using 6-OHDA or the beta-receptor antagonist nadolol alters the typical pattern in production of T helper 1 (Th1) and Th2 cytokines seen in cultures of spleen cells from C57BL/6 mice infected with murine AIDS (MAIDS). We found that neither method of sympathetic blockade affected cytokine response to MAIDS. We also found that the norepinephrine concentration and content of the spleen were reduced dramatically by the MAIDS infection itself at 3 and 6 weeks after LP-BM5 inoculation. This finding has not been previously reported in mice with MAIDS and suggests that the viral infection itself produces a functional sympathectomy in the spleen, a target of that infection. PMID- 11908923 TI - Transforming growth factor-beta 1 and cortisol in differentially reared primates. AB - Exposure of primate infants to adverse rearing conditions during the first half year of life can result in enduring behavioral, neuroendocrine, and immunologic abnormalities. However, the effects of differential rearing on cytokines, some of which can regulate immune and inflammatory responses and modulate activity of the central nervous system and the hypothalamic-pituitary-adrenal (HPA) axis, are largely unexamined. The present study explored the relationship between circulating levels of transforming growth factor-beta 1 (TGF-beta 1) and cortisol in macaques reared either normally or under conditions of variable foraging demand (VFD). Under VFD rearing, for a period of 4 months, the infants' mothers intermittently had to expend more time and effort to obtain food than did the mothers of normally reared control subjects. Two years after cessation of the rearing experience, exposure to a moderate stressor (confinement in an unfamiliar room for 90 min) induced elevated levels of serum TGF-beta 1 and plasma cortisol in VFD-reared monkeys compared to normally reared controls. The correlation between TGF-beta 1 and cortisol levels was substantially higher in the normally reared subjects. Examination of the relationship between HPA axis and immune function will improve our understanding of the pathophysiological consequences of adverse rearing. PMID- 11908924 TI - Altered kinetics of IL-1 alpha, IL-1 beta, and KGF-1 gene expression in early wounds of restrained mice. AB - Inflammatory processes that occur after injury contribute to wound closure. Previous studies showed that wounds of restraint-stressed (RST) mice had a reduced number of inflammatory cells and healed more slowly compared to controls. To investigate the molecular mechanisms between stress and wound healing, we studied cutaneous gene expression of IL-1 alpha, IL-1 beta, and KGF-1. Female SKH 1 mice were restrained for 3 days before and for 5 days following placement of cutaneous wounds. Wounds were subjected to competitive RT-PCR. At day 1, RST mice had significantly lower IL-1 beta and KGF-1 mRNA than controls. At day 5, RST mice had significantly higher IL-1 alpha and IL-1 beta mRNA than controls. Treatment of RST mice with the glucocorticoid receptor antagonist RU486 restored IL-1 beta mRNA expression at day 1. Our results suggest that stress induces alterations in the kinetics of cutaneous proinflammatory cytokine and growth factor gene expression which could impair the quality of healing tissues. PMID- 11908925 TI - New molecular targets and biological therapies in sarcomas. AB - The treatment of patients with soft tissue and bone sarcomas has dramatically improved over the last decade. This improvement has been brought about through advances in diagnosis, surgical techniques, conformal radiotherapy, and combination chemotherapy. Further advances in the management of the diverse spectrum of sarcoma patients will reflect tailoring of therapy based on molecular abnormalities. The role of cytogenetics and molecular analysis of fusion or mutated genes in diagnosis, prognosis, and design of biological treatments is discussed. An example of this approach has been the recent success in treatment of patients with gastrointestinal stromal tumours expressing mutant c-kit with a specific tyrosine kinase inhibitor, STI571. Molecular rearrangements may also serve as targets for designing specific immunotherapies with the fusion gene product. The use of biological therapies with signal transduction inhibitors, angiogenesis inhibitors, matrix metalloproteinase inhibitors, immunotherapy, differentiation inducers, and gene therapy could complement existing treatments for long-term control of disease. As these newer biological agents take form, clinical trial design will undergo change to reflect the chronic nature of these therapies. PMID- 11908926 TI - Remission induction therapy for childhood acute lymphoblastic leukaemia: clinical and cellular pharmacology of vincristine, corticosteroids, L-asparaginase and anthracyclines. AB - Remission induction therapy with vincristine, a corticosteroid, L-asparaginase and an anthracycline has been the mainstay of the initial phase of treatment for childhood acute lymphoblastic leukaemia (ALL) for the past 25 years. The speed and depth of the early response to remission induction therapy has become an important determinant of the intensity of subsequent therapy in many protocols worldwide. Moreover, the detection of significant levels of minimal residual disease at the end of remission induction may have an important bearing on subsequent outcome. Although these clinical observations may reflect, in part, the inherent sensitivity of lymphoblasts to remission induction therapy, the pharmacology of these agents in relation to childhood ALL may also play an important part in early response to therapy. In-vitro studies of human leukaemia cell lines indicate that both the extracellular fluid concentration and duration of exposure to vincristine and anthracyclines are important determinants of cytotoxicity. For L-asparaginase and corticosteroids, the cellular and molecular pharmacological determinants of chemosensitivity have been partially characterized, but further work is needed in this area. The clinical pharmacology of vincristine and L-asparaginase have been well characterized in relation to childhood ALL, and considerable interpatient pharmacokinetic variability exists for these drugs. For corticosteroids and anthracyclines, pharmacology studies are needed in order to fully characterize and understand the factors influencing interpatient pharmacokinetic variability for these agents in relation to childhood ALL. Whereas the relationship between the clinical pharmacology, and potentially important pharmacodynamic effects such as asparagine depletion, has been well characterized for therapy with L-asparaginase, similar studies have yet to be performed for the other drugs that form the mainstay of remission induction therapy for childhood ALL. Therefore, further studies are required to investigate the relative importance of the clinical and cellular pharmacology of vincristine, corticosteroids, L-asparaginase and anthracyclines in the speed and depth of response to remission induction therapy for childhood ALL. Where these have been studied, interindividual differences in the clinical and cellular pharmacology of anticancer agents have been shown to be important determinants of the long-term disease-free survival for children with ALL. PMID- 11908927 TI - Consolidation therapy for childhood acute lymphoblastic leukaemia: clinical and cellular pharmacology of cytosine arabinoside, epipodophyllotoxins and cyclophosphamide. AB - The intensification of post-remission induction therapy has been shown to improve the relapse-free survival for childhood acute lymphoblastic leukaemia (ALL), and is now a standard component of the treatment of childhood acute lymphoblastic leukaemia. For cytosine arabinoside (ara-C), methotrexate, vincristine and corticosteroids, in-vitro studies indicate that the extracellular drug concentration and exposure time are important determinants of cytotoxicity for human leukaemia cell lines. For L-asparaginase, epipodopyllotoxins and cyclophosphamide, there have been few studies of the relationship between cellular pharmacology and cytotoxicity in relation to ALL. The clinical and cellular pharmacology of methotrexate and cytosine arabinoside have been studied in relation to childhood ALL in vivo. For these drugs, there is evidence to suggest that maintenance of plasma concentrations that are biochemically optimal is necessary to maximize anti-leukaemic effects. For cytosine arabinoside in particular, optimal extracellular fluid concentrations are not likely to be achieved or maintained by bolus or short-duration i.v. infusions. A potentially important example of this may be served by the success of antimetabolite-based intrathecal chemotherapy for CNS-directed treatment of childhood ALL. Intrathecal administration of both methotrexate and cytosine arabinoside results in prolonged leukaemic cell exposure to cytotoxic concentrations of the drug. For vincristine, anthracyclines and asparaginase, the actual dose intensity received by children during consolidation therapy may be important, and there is considerable interpatient variation in the pharmacokinetics of cyclophosphamide and teniposide in the therapy of childhood cancers. The importance of this relationship to childhood ALL is not known. The pharmacological and cellular pharmacological studies performed at St Jude Children's Research Hospital (Memphis, TN, USA) have allowed investigation of the relationships between the clinical and cellular pharmacology of methotrexate and prognosis, and have supported the individualization of consolidation therapy with this drug. Cytosine arabinoside has been less well studied in relation to childhood ALL, although evidence exists to suggest that the administration of conventional-dose bolus or infusion schedules may not be optimal in terms of the antileukaemic efficacy of this antimetabolite. For L-asparaginase, ongoing studies may allow the relationship between dose and schedule of administration to be related to pharmacodynamic measures such as asparagine depletion and prognosis. Therefore, through knowledge of clinical and cellular pharmacological properties, it may be possible to optimize the consolidation phase of therapy for childhood ALL, without disrupting the fundamental principles by which the overall treatment is administered. This may be particularly important for children with disease that has inherent or acquired resistance to therapy. PMID- 11908928 TI - Continuing therapy for childhood acute lymphoblastic leukaemia: clinical and cellular pharmacology of methotrexate, 6-mercaptopurine and 6-thioguanine. AB - Across the world, therapy with 6-mercaptopurine (6-MP) and methotrexate (MTX) forms the basis of the continuing therapy of childhood acute lymphoblastic leukaemia (ALL). In this review, the pharmacological determinants of the sensitivity of human leukaemia cell lines and lymphoblasts derived from children with ALL will be discussed. In addition, clinical pharmacological studies of 6-MP and MTX in relation to the continuing therapy with childhood ALL will be reviewed. For 6-MP in vitro, prolonged exposure times to relatively high extracellular drug concentrations are necessary for cytotoxicity, and these concentrations are much higher than those achieved during continuing therapy for childhood ALL. For MTX, plasma concentrations are achieved during continuing therapy that would be cytotoxic to human leukaemia cells during prolonged exposures in vitro. For both MTX and 6-MP, wide inter- and intrapatient variation in plasma pharmacokinetic parameters has been described. For 6-MP and MTX, cellular pharmacological studies have been largely restricted to erythrocytes as a surrogate of the possible effects in leukaemic blasts. Although measures of the pharmacology of 6-MP and MTX in erythrocytes has been related to prognosis in many studies, 6-MP systemic exposure and the dose intensity of 6-MP and MTX actually received by children during this phase of therapy seems to be the most important determinant of efficacy. Further studies will be needed to determine the importance of pharmacokinetic variability during continuing therapy as a determinant of outcome for children with ALL. In this respect, minimal residual disease status during this phase of treatment may prove to be a useful pharmacodynamic endpoint. PMID- 11908929 TI - Rationale and techniques of intra-operative hyperthermic intraperitoneal chemotherapy. AB - BACKGROUND: In recent years surgical cytoreduction followed by intra-operative hyperthermic intraperitoneal chemotherapy (HIPEC) was introduced as treatment modality in patients with peritoneal surface malignancy. In the current review the rational for this approach, the prerequisites and the different techniques used are discussed. METHODS: A literature search through PubMed was performed. RESULTS: Pharmacokinetic studies have shown an important dose advantage for intraperitoneal versus intravenous application. Hyperthermia enhances the penetration of cytostatic drugs into tumour tissue and also shows synergism with various cytostatic drugs. The penetration depth of drugs into tissue is limited, therefore HIPEC can only be effective in patients with minimal residual disease after (aggressive) surgery. HIPEC can be conducted in various ways, without clear proven advantage of one method over the others. Local complications after this combined treatment approach are mainly surgery related. Intraperitoneal chemotherapy may cause systemic toxicity, dependent on the drug used. In randomised studies cytoreductive surgery followed by HIPEC has proven its value in the prevention of peritoneal dissemination in gastric cancer. Phase II data on HIPEC in peritoneal carcinomatosis of colorectal origin and pseudomyxoma peritonei are promising, but randomised studies are still not available. CONCLUSION: Aggressive surgical cytoreduction and HIPEC in patients with peritoneal surface malignancy has a clear rational and seems to have clinical value. PMID- 11908930 TI - Chemo-immunotherapy and chemo-adoptive immunotherapy of cancer. AB - The chemo-immunotherapy (CIT) and chemo-adoptive immunotherapy (CAIT) regimens tested in the past decade are summarized. From them we have learned a great deal about the interactions between various chemotherapeutic agents, immune modulating agents and effector cells. The most commonly reported result in multi-modality experiments with CAIT has been a synergistic enhancement in antitumor activity. Clinical trials usually demonstrated improvement in patient quality of life, an extension of survival time, and occasional complete regression of tumor. In many animal models, this enhancement often meant the complete regression and apparent cure of tumor in the animal. One mechanism by which this synergistic enhancement takes place appears to be a suppression of tumor-associated suppressor T cell activity by the chemotherapeutic agents, thereby inducing enhanced cytolytic activity against tumor by the adoptively transferred, activated effector cells. In CAIT the most commonly used drug has been cyclophosphamide. In CIT a wide variety of chemotherapy agents have been used but none of the clinical trials made use of cyclophosphamide. Thus, direct comparisons are not possible. Suggestive of the intricate regulatory processes involved, many CIT studies indicate a synergy only when specific doses of chemotherapy and immunotherapy agents are given, and in a specific sequence. CIT has become less toxic, is being handled on a cost-effective outpatient basis, while maintaining similar objective response rates to earlier inpatient treatments. In the future, CAIT and CIT will probably have an increasing role in the management of patients with specific cancers. PMID- 11908931 TI - Childhood asthma hospitalization and residential exposure to state route traffic. AB - This study investigated whether pediatric hospitalization for asthma was related to living near a road with heavy traffic. In this case-control study, cases (N=417) consisted of white children aged 0-14 years who were admitted for asthma and who resided in Erie County, New York, excluding the city of Buffalo. Controls (N=461) were children in the same age range admitted during the same time period for nonrespiratory diseases. Subjects' residential addresses were linked to traffic information provided by the New York State Department of Transportation. After adjustments for age and poverty level were made, children hospitalized for asthma were more likely to live on roads with the highest tertile of vehicle miles traveled (VMT) (odds ratio (OR): 1.93, 95% confidence interval (CI): 1.13 3.29) within 200 m and were more likely to have trucks and trailers passing by within 200 m of their residence (OR=1.43, 95% CI: 1.03-1.99) compared to controls. However, childhood asthma hospitalization was not significantly associated with residential distance from state roads, annual VMT within 500 m, or whether trucks or trailers passed by within 500 m. This study suggests that exposure to high volumes of traffic/truck within 200 m of homes contributes to childhood asthma hospitalizations. PMID- 11908932 TI - Sociodemographic, clinical, and laboratory features of cases of organic phosphorus intoxication who attended the Emergency Department in the Southeast Anatolian Region of Turkey. AB - In this study, sociodemographic, laboratory, and clinical features of cases of organic phosphorus (OP) intoxication in the Southeast Anatolian region of Turkey were investigated. Patients with OP intoxication admitted to the Emergency Service of Dicle University Hospital in Diyarbakyir City between May and August 1998 were evaluated. This prospective cohort study included five male (M) and 19 female (F) consecutive patients. Five cases were accidental intoxication; however, 19 cases were suicide attempts. Mean age of cases was 24+/-11 years; 54.2% of them were under the age of 20 years and 83.3% of them were under the age of 30 years. The M/F ratio was 1.0/3.8. The cases of suicidal purposeful intoxications were mostly determined in singles (58.3%, P<0.05). Most of the cases had a primary education level (16, 66.7%) and a lower socioeconomic status (14, 58.3%); 79.2% of cases admitted to our emergency service received atropine in primary health care centers. In emergency service, pralidoxime (PAM) was administered to only 29.2% of cases. According to ECG examination, tachycardia (14, 58.3%), ST changes (13, 54.2%), and T changes (3, 12.5%) were mostly seen; bradycardia and serious ventricular arrhythmias were not seen in any case. Patients who received atropine plus PAM had higher rates of arrhythmias, but this was not statistically significant (P>0.05). The most observed biochemical features of cases were leukocytosis (21 cases), hypokalemia (18 cases), and hyponatremia (4 cases). Other biochemical features were not evidently altered. In present cases, the most frequently seen symptoms and findings were vomiting (18, 75%), feeling faint (17, 70.8%), and tachycardia and dozing off (14, 58.3%). Serious ventricular arrhythmias were not observed, and in our region, OP intoxication especially affected young unmarried females, and most of them resulted from a suicidal purpose. PMID- 11908933 TI - Acute accidental exposure to chlorine gas in the Southeast of Turkey: a study of 106 cases. AB - The present study reports a thorough investigation of the sociodemographic characteristics, clinical findings, and treatment of persons affected acutely by chlorine gas exposure from a chlorine tank belonging to the municipality of Diyarbakir. One hundred six persons were assessed. In this cross-sectional study, 58 patients were male and 48 were female. Children and adolescents younger than 18 years constituted more than half of the patients (60 cases, 56.6%). The age of patients ranged between 3 months and 75 years. Among the cases evaluated in emergency rooms, 7 patients had mild poisoning and were discharged after first examinations and symptomatic treatments, 62 patients were moderately affected and were taken under observation, and the remaining 37 were severely affected and were hospitalized. In physical examinations, 29 patients had expiratory wheezing, and 1 had tachycardia and extrasystoles. There were no deaths among these patients, acute chlorine intoxication affected mostly children. Respiratory tract findings were predominant in most of the patients. Steroid and bicarbonate applications were inadequate supportive therapies. Humidified O(2) and beta agonist applications were most useful in the therapy of acute chlorine intoxication. PMID- 11908934 TI - Utility of the WHO neurobehavioral core test battery in Chinese workers-a meta analysis. AB - We performed a meta-analysis to determine the most sensitive subtests of the WHO Neurobehavioral Core Test Battery (NCTB) when administered to Chinese workers. Extensive Chinese Biological Medical Database and MEDLINE searches, review of cited references, and discussion with other investigators were undertaken. Data were extracted from 39 eligible studies; the summary effects (effect sizes) were calculated using a fixed-effect model. Various exposure agents showed different sensitivities to the seven subtests of NCTB. For mercury-exposure, the Benton Visual Retention was the most sensitive subtest, with an effect size (95% CI) of 6.0 (4.4-7.6). For lead-exposure, the most sensitive subtests were the Pursuit Aiming II and Profile of Mood States, with effect sizes (95% CI) of 11.3 (8.3 14.3) and 10.6 (7.5-13.7), respectively. For organic solvents-exposure, Digit Span, Pursuit Aiming II, and Digit Symbol were the most sensitive subtests, with effect sizes (95- CI) of 4.7 (3.3-6.1), 4.6 (3.1-6.1), and 4.1 (2.7-5.5), respectively. PMID- 11908935 TI - Interleukins 2 and 12 produce significant recovery of cytotoxic function in dibutyltin-exposed human natural killer cells. AB - Cytotoxic function of human natural killer (NK) cells is modulated by a variety of cytokines. Interleukins (IL) 2, 12, 15, and 18 and Interferon gamma (IFNgamma) are potent stimulators of NK cell cytotoxicity. Butyltins (BTs) are used in a variety of consumer products and industrial applications. Dibutyltin (DBT) is found in plastic products, beverages stored in PVC pipes during manufacturing, and poultry products. BTs appear to increase the risk of cancer and viral infections in exposed individuals. Recently, we have demonstrated that the ability of NK cells to kill tumor cells is greatly diminished after a 1-h exposure to dibutyltin. This inhibition of tumor killing function continues even after removal of the compound. There is no significant recovery of NK cytotoxic function even when the cells are allowed to recover for 6 days. In the current study we examine the effects of NK-stimulatory cytokines on the ability of NK cells to recover from the inhibitory effects of a 1-h DBT treatment. Highly purified NK cells (>95% CD16(+)) or a lymphocyte preparation containing both T lymphocytes and NK cells were treated with 5 microM DBT and then allowed to recover for 24 h, 48 h, 4 days, and 6 days in DBT-free medium containing either no cytokine or a maximally stimulatory dose of several NK-stimulatory cytokines. Tumor killing function was tested using a radioactive chromium release assay. As seen in our previous studies there is no recovery of NK cell cytotoxic function even after a 6-day recovery period when no cytokine is present in the medium. However, there is significant recovery of NK cytotoxic function when IL2, IL12, or the combination of IL2 plus IL12 is present in the medium during the recovery period. The other cytokines tested (IL15, IL18, and IFNgamma) were unable to increase the cytotoxicity of DBT-exposed NK cells. PMID- 11908936 TI - Variation of systolic blood pressure in rats exposed to cadmium and nickel. AB - This research was intended to verify separate and combined effects of cadmium and nickel on blood pressure in rats. After cadmium chloride (CdCl(2)) and nickel chloride (NiCl(2)) were administered individually and combined by intraperitoneal injection to rats daily for a week, systolic blood pressure of the tails were measured at 1, 5, 10, 20, and 30 days after administration. Each substance was injected into rats with 0.1 mg/kg x bw and 1.0 mg/kg x bw concentrations. After 0.1 mg/kg x bw CdCl(2) was injected, statistically significant differences from the control group (only saline) were found at 1, 5, and 10 days. After 0.1 mg/kg x bw NiCl(2) was injected, no statistically significant differences from the control group were found. After 0.1 mg/kg x bw CdCl(2) and 0.1 mg/kg x bw NiCl(2) were injected combined simultaneously, statistically significant differences from the control group were found at 1, 5, and 10 days, compared with the 0.1 mg/kg x bw CdCl(2) group after 5 days and the 0.1 mg/kg x bw NiCl(2) group after 5 and 10 days. After 1.0 mg/kg x bw CdCl(2) was injected, statistically significant differences from the control group were found at 1, 5, 10, and 20 days. After 1.0 mg/kg x bw NiCl(2) was injected, statistically significant differences from the control group were found at 1 and 5 days. After 1.0 mg/kg x bw CdCl(2) and 1.0 mg/kg x bw NiCl(2) were injected combined, statistically significant differences were found at 1, 5, 10, 20, and 30 days, compared with 1.0 mg/kg x bw CdCl(2) at 10, 20, and 30 days and 1.0 mg/kg x bw NiCl(2) at 5, 10, 20, and 30 days. It was found that the effect of CdCl(2) on blood pressure was much more than that of NiCl(2) and the combination of CdCl(2) and NiCl(2) in high concentration delayed recovery of blood pressure. PMID- 11908937 TI - Exposure of children to lead and cadmium from a mining area of Brazil. AB - During the past 50 years the Ribeira river valley, in the southern part of the state of Sao Paulo, Brazil, had been under the influence of the full activity of a huge lead refinery and mine working by the side of the river. The plant completely stopped all kinds of industrial activities at the end of 1995, and part of the worker population and their families still remain living nearby in small communities. The objective of the study was to assess the exposure of children to lead and cadmium in these areas, where residual environmental contamination from the past industrial activity still exists. Blood samples of 295 children aged 7 to 14 years, residing in rural and urban areas around the mine and the refinery, were collected. A questionnaire was given to gather information on food habits, leisure activities, father's past employment, current and former residential places, and other variables. Blood lead and cadmium concentrations were analyzed by graphite furnace atomic absorption spectrometry using Zeeman background correction. Cadmium values obtained in this population were mostly below established quantification limits (0.5 microg/dl). The median of blood lead level (BLL) obtained in children living close to the lead refinery was 11.25 microg/dl, and the median in other mining regions far from the refinery was 4.4 microg/dl. Logistic regression analysis was conducted to examine the independent contribution of selected variables in predicting BLL in these children. The following variables showed significant association with high BLL: residential area close to the lead refinery [odds ratio (OR)=10.38 (95% confidence interval (Cl)=4.86-23.25)], former father's occupational lead exposure [OR=4.07 (95% Cl=1.82-9.24)], and male gender [OR=2.60 (95% Cl=1.24-5.62)]. PMID- 11908938 TI - Butyltin compounds in human liver. AB - Intake of marine food is the main source of butyltin exposure in humans. Health effects following exposure to butyltin compounds are usually in the immune system, but endocrine effects of butyltin from a variety of marine species have been documented. The information on human exposure to butyltin compounds and hepatic deposition is limited. The present study include 18 consecutively sampled human livers analyzed for butyltin compounds. Dibutyltin (DBT) concentrations varied between 0.8 and 28.3 ng/g with a mean concentration of 9.0 ng/g. Significantly lower concentrations of monobutyltin (MBT) were observed, ranging from 0.3 to 4.7 ng/g with a mean value of 1.6 ng/g. Age and DBT/MBT ratio were significantly associated. We suggest that younger men have more recent exposures or have a lower capacity to debutylate DBT than older men and would therefore potentially be more susceptible to butyltin toxicity. Given the interperson variability observed in our limited group of men, we cannot exclude that thresholds for either immunotoxicity or effects on the endocrine system may occur due to exposure to butyltin compounds alone or in combination with other environmental toxicants with similar target organs. PMID- 11908939 TI - Dichlorodiphenyltrichloroethane in soil, river sediment, and fish in the Amazon in Brazil. AB - Dichlorodiphenyltrichloroethane (DDT), its main metabolites, and other organochlorines were analyzed in soils (n=6), fluvial sediments (n=14), and fish (n=10) that were collected in several areas of the Amazon region in Brazil. The samples were analyzed by capillary column gas chromatography coupled to electron capture detection. DDT residues were present in most of the collected sediments in concentrations of approximately 10 to 100 micro/kg (ppb, dry weight). Some urban top soils were found to have more than 1 mg/kg (ppm). In fish, as much as 0.5 mg/kg of total DDT (wet weight) was found in the edible parts. The presence of p,p'-DDT in most of the samples reflects the use of this insecticide against vectors of malaria between 1946 and 1993, which has led to its ubiquitous presence in the environment of the Brazilian Amazon. PMID- 11908941 TI - CD40/CD40L interaction is essential for the induction of EAE in the absence of CD28-mediated co-stimulation. AB - CD28 provides a co-stimulatory signal critical for optimal T cell activation. We and others have shown that the B7/CD28 co-stimulatory pathway is a major regulatory pathway for the control of immune responses. Experimentally induced models of autoimmunity have been shown to be prevented or reduced in intensity in mice deficient for CD28. Here, we show that EAE and accompanying neuroantigen specific immune responses are drastically reduced in the absence of CD28. However, we go on to show that EAE can be induced in CD28-deficient mice following two immunizations. After re-immunization, CD28-deficient mice develop severe EAE with myelin-specific responses equal to those of wildtype controls, and extensive demyelination in the spinal cord. Treatment of CD28-deficient mice with anti-CD40L at the time of immunization significantly reduced DTH responses and prevented the development of EAE following two immunizations, indicating a critical role for CD40/CD40L signaling in the absence of CD28. Taken together, our results indicate that CD28-mediated co-stimulation does not regulate immunological anergy. Instead, CD28 appears to adjust the threshold for activation and expansion of autoreactive cells. PMID- 11908942 TI - The influence of HLA-DR4 (0401) on the immune response to type II collagen and the development of collagen induced arthritis in mice. AB - Rheumatoid arthritis (RA) is an autoimmune disease that is genetically associated with the MHC class II molecule HLA-DRbeta1*0401 (DR4). In order to determine if this MHC can influence the immune response to the candidate autoantigen type II collagen (CII), we have studied collagen induced arthritis (CIA) resistant C57BL/6 mice, made transgenic (Tg) for human DR4. These DR4 Tg mice exhibited a strong T cell proliferative response to CII and its DR4 restricted peptide p261 273 after immunization with these antigens that was not seen in the C57BL/6 wild type mice. DR4 Tg mice also exhibited an increase in IFN-gamma production in response to CII, indicating the activation of Th1 cells. While these Tg mice produced IgM anti-CII antibodies, they failed to produce a detectable level of IgG2a (Th1 type) anti-bCII antibody and did not develop CIA. This study shows that a Th1 type T cell response to CII can be established in CIA non-susceptible mice by introducing the human transgene, DR4. This T cell response, however, is not sufficient to induce an antibody isotype switch to IgG2a, nor is it sufficient for the induction of CIA. These results may help to explain why many individuals expressing HLA-DRbeta1*0401 do not develop RA. PMID- 11908943 TI - Absence of CD5 dramatically reduces progression of pulmonary inflammatory lesions in SHP-1 protein-tyrosine phosphatase-deficient 'viable motheaten' mice. AB - Mice homozygous for the viable motheaten (Hcph(me-v)) mutation are deficient in SHP-1 protein-tyrosine phosphatase, resulting in severe systemic autoimmunity and immune dysfunction. A high percentage of B-cells in viable motheaten mice express the cell surface glycoprotein CD5, in contrast to wild type mice that express CD5 on only a small percentage of B-cells. CD5(+) B-cells have been associated with autoantibody production. To determine the role of CD5 in the development of the inflammatory disease in me(v)/ me(v) mice, we created a stock of CD5(null)me(v)/ me(v) mice. The longevity of CD5(null)me(v)/ me(v) mice was increased 69% in comparison to me(v)/ me(v) mice on a similar (B6;129) background. The increased lifespan was associated with a marked reduction in pulmonary inflammation. Flow cytometry analysis of spleen cells from CD5(null)me(v)/ me(v) mice at 9-12 weeks of age revealed significant decreases in percentages of IgM/B220 double positive B-cells, Mac-1/Gr-1 double positive cells and CD4(+) T-cells compared with me(v)/ me(v) mice. CD5(null)me(v)/ me(v) mice also had significantly lower serum IgM levels in comparison to me(v)/ me(v) mice. Study of CD5(null)me(v)/ me(v) mice may provide further insight into the role of CD5 in cell signaling and may help explain the observed association of CD5(+) B-cells with autoimmune disease. PMID- 11908944 TI - T cell developmental defects in 'viable motheaten' mice deficient in SHP-1 protein-tyrosine phosphatase. Developmental defects are corrected in vitro in the presence of normal hematopoietic-origin stromal cells and in vivo by exogenous IL 7. AB - Defects in the gene that encodes SHP-1 protein tyrosine phosphatase result in multiple hematopoietic abnormalities and generalized autoimmunity in viable motheaten (me(v)) mice. These mice also exhibit early thymic involution and abnormalities in T cell development. Here, we describe the use of fetal thymic organ culture (FTOC) and bone marrow adoptive transfer to study the effects of SHP-1 deficiency on thymocyte development. Chimeric FTOC established with normal bone marrow placed onto deoxyguanosine-treated fetal thymic lobes or onto scid fetal thymic lobes generated T cells. Bone marrow from SHP-1-deficient me(v)/ me(v) mice generated decreased numbers of T cells in chimeric FTOC established using deoxyguanosine-treated thymi but generated normal numbers in chimeric FTOC established using scid thymi. However, scid fetal thymi seeded with me(v)/ me(v) bone marrow also exhibited morphological abnormalities and contained elevated numbers of macrophages. Addition of IL-7 to me(v)/ me(v) bone marrow-seeded scid FTOC led to increased cell numbers, particularly of macrophages. Intrathymic injection of IL-7 partially restored the ability of progenitor cells in me(v)/ me(v) bone marrow to populate the thymus of adoptive recipients. We conclude that abnormal T cell development in me(v)/ me(v) mice may in part be due to defects in the ability of bone marrow-derived accessory cells to provide bioavailable IL-7 to developing thymocytes. PMID- 11908945 TI - Experimental chronic Chagas' disease myocarditis is an autoimmune disease preventable by induction of immunological tolerance to myocardial antigens. AB - The protozoan Trypanosoma cruzi causes chronic Chagas' disease myocarditis (CCDM) in infected mammals. The pathogenesis of CCDM, however, is still unclear. Indirect evidence for either parasite- or heart-specific immune responses playing a pathogenic role is available. In this work, the participation of autoimmunity in the development of CCDM is demonstrated in mice in which immunological tolerance to heart antigens was induced or strengthened prior to their infection by T. cruzi. Tolerance was induced by heart antigen administration in the presence of complete Freund's adjuvant and anti-CD4 antibodies. Tolerized mice developed less intense CCDM than control non-tolerized animals that had received only anti-CD4 and adjuvant. This result confirms the important notion that tolerance to self, and in particular to heart antigens, may be reinforced/induced in normal animals, and raises the possibility that analogous interventions may prevent the development of CCDM in millions of T. cruzi -infected human beings. PMID- 11908947 TI - Characterization of the dominant autoreactive T-cell epitope in spontaneous autoimmune haemolytic anaemia of the NZB mouse. AB - NZB mice spontaneously develop autoimmune haemolytic anaemia (AIHA) due to a T helper-dependent autoantibody response against the erythrocyte anion channel protein, Band 3. Here, we characterize the recognition of the Band 3 sequence 861 874, which carries the dominant, I-E(d)-restricted T cell epitope. The ability of N and C-terminal truncated versions of peptide 861-874 to elicit NZB splenic T cell proliferation indicated that the core epitope spans residues 862-870. Next, a set of alanine substitution analogues was tested to determine which residues functioned either as MHC anchor or TCR contact residues. A combination of proliferation and MHC:peptide binding assays identified residues 862(L), 864(V), 865(L), and 869(K) as I-E(d) anchor residues, and 868(V) as the only TCR contact residue. The ability of the wild-type sequence 861-874 to compete with a high affinity reference peptide for binding to I-E(d) indicates that the escape of pathogenic NZB T cells from purging of the autoreactive repertoire cannot be attributed to ineffective presentation of peptide 861-874 by its restricting element. It will now be possible to design altered peptide ligands of Band 3 861 874, in order to further dissect the mechanisms responsible for the maintenance and loss of T cell tolerance to RBC autoantigens, and to modulate the immune response in AIHA. PMID- 11908946 TI - Cross-reactive mycobacterial and self hsp60 epitope recognition in I-A(g7) expressing NOD, NOD-asp and Biozzi AB/H mice. AB - The highly conserved 60 kD endogenous heat shock protein (hsp60) has been suggested to be a target for T cell recognition in autoimmune diseases such as type I diabetes. We previously reported cross-recognition of both mycobacterial hsp60 (Mt60) and self hsp60 (m60) by Mt60 immunized NOD mice. To identify the epitopes involved, we generated T cell lines against m60 or its mycobacterial counterpart and tested these lines for recognition of complete sets of overlapping peptides spanning either hsp60 sequence. T cell lines responded to identical regions in the hsp60 proteins, regardless of their degree of conservation or I-A(g7) binding-affinity. Additionally, we determined whether a protective genetic background would affect the presence of hsp60 cross-reactive T cells in the peripheral repertoire by comparing epitope recognition in I-A(g7) expressing NOD, NOD-asp and Biozzi AB/H mice. Two out of five immunodominant murine peptides were able to induce proliferation in NOD and NOD-asp Mt60 T cell lines, but not in Biozzi AB/H T cell lines. Our results point out that Mt60 immunization not necessarily leads to proliferative T cells responding to endogenous hsp60 peptides in the context of diabetes-predisposing I-A(g7). Moreover, the capacity of T cells to respond to self hsp60 is not influenced by the presence of protective I-A(g7asp). Yet, proliferation of hsp60 autoreactive T cells is solely measured in combination with insulitis and as such serves as a surrogate marker for islet inflammation. PMID- 11908948 TI - Gene-expression profile of collagen-induced arthritis. AB - To provide a global analysis of genes involved in the inflammatory process in joints of DBA/1J mice suffering from collagen induced arthritis (CIA) we used oligonucleotide microarrays representing approximately 11,000 genes to determine the gene expression profile of the inflamed paws at peak of disease, and compared them to normal tissue. Peak of disease was determined from clinical evaluation of disease and histopathology of joints. Of the 11,000 genes assayed, 223 showed differential expression of four fold or more (187 upregulated and 36 downregulated). Ninety-five of the genes observed had well-characterized full length sequences in databases, and 128 were unknown (Ests). Inflammation resulted in a profile of increased gene expression of matrix metalloproteinases, immune related, extra-cellular matrix and cell adhesion molecules, as well as molecules involved in cell division and transcription; differential regulation of molecules involved in signal transduction, protein synthesis and metabolism. Of the 55 genes with known chromosomal locations nine mapped to previously identified QTL, contributing to susceptibility or severity of CIA, i.e. MHC class I, II, Basigin, FAP, Cathepsin K, CD 53, RAF1, glucagon, and retinal taurine transporter. The profile of gene expression supports current theoretical models of disease progression and might open new perspectives for both diagnosis and treatment of arthritis. PMID- 11908949 TI - An organic CD4 inhibitor reduces the clinical and pathological symptoms of acute experimental allergic encephalomyelitis. AB - CD4(+) T cells have an important role in mediating the pathogenesis of many human and experimental autoimmune diseases including experimental allergic encephalomyelitis (EAE), a demyelinating animal model for multiple sclerosis (MS). We applied a computer screening approach to select a small organic molecule, TJU103, that would specifically inhibit autoreactive CD4(+) T cells by disrupting the function of the CD4 molecule during activation. Upon studying the therapeutic effect of TJU103 in acute EAE, it was found that administration shortly before or after the onset of clinical symptoms reduced the severity of disease in both SJL and SWXJ-14 mouse models. In addition, TJU103 treatment could affect both in vivo responses to EAE rechallenge and secondary in vitro proliferation and cytokine production of T cells responding to proteolipid protein epitope 139-151 (PLPe). These results demonstrate the potential of the TJU103 organic inhibitor for future clinical application in CD4(+) T cell mediated diseases. PMID- 11908950 TI - Characterization of anti-phosphatidylcholine polyreactive natural autoantibodies from normal human subjects. AB - In 1990 our group reported a patient with autoimmune hemolytic anemia and high titers of IgM anticardiolipin antibodies that cross-reacted with phosphatidylcholine (PTC). These autoantibodies also recognized bromelain-treated erythrocytes (BrE) and in vitro aged erythrocytes. The epitope exposed with this treatment is PTC. To detect and characterize antiphosphatidylcholine antibodies (anti-PTC) in a normal human population, we studied by ELISA the presence of serum anti-PTC (IgG and IgM) in clinically healthy human subjects. The most representative samples were also studied for IgG or IgM activity against BrE by flow cytometry, rheumatoid factor activity, anti-dsDNA, anti-ssDNA by ELISA and by indirect immunofluorecence (IIF) using HEp-2 line and a healthy human fibroblast strain as substratum. Eighty five percent of sera had IgM anti-PTC and none had IgG. IgM antibodies against BrE were inhibited by PTC micelles (mPTC). Anti-PTC were also inhibited by phosphorylcholine and phosphatidic acid. Aggregated gammaglobulin (AGG) reactivity was inhibited by dsDNA and mPTC. The IgM anti-dsDNA activity was inhibited by soluble dsDNA, AGG and mPTC. All sera gave intermediate filaments pattern by IIF and reacted against purified vimentin by dot blot and Western blot.Our study shows hemolytic IgM anti-PTC present in normal human serum. The main epitope recognized by these autoantibodies is phosphorylcholine. The physicochemical characteristics, crossreactivity with self antigens and functional properties are typical features of natural autoantibodies. PMID- 11908952 TI - Fission-track zircon age of the Eocene Pondaung Formation, Myanmar. PMID- 11908951 TI - ICA512(IA-2) epitope specific assays distinguish transient from diabetes associated autoantibodies. AB - ICA512/IA-2, a tyrosine phosphatase-like protein, is one of the major autoantigens in type 1 diabetes. Following phage display characterization of ICA512 autoantigenic epitopes, we developed fluid phase autoantibody radioimmunoassays for a series of ICA512 fragments (F1 [amino acids (aa): 761 964], F2A [aa 256-760], F2B [aa 761-928], and F2C [aa 929-979]). With the hypothesis that 'non-diabetes associated' ICA512 autoantibodies would differ from diabetes associated ICA512 autoantibodies in terms of epitopes recognized, we analyzed ten such serum samples (two from normal control individuals, one from a general population subject with transient ICA512 autoantibodies and seven from relatives of patients with type 1 diabetes who had single transient ICA512 positivity). All but one of the 'non-diabetes associated' ICA512 positive samples (9/10) did not react with Fragment 1 which contains the major antigenic epitopes of the molecule that were recognized by almost all (51/52) ICA512 positive new onset patient samples and pre-diabetic relatives (P< 10(-6)). The great majority of samples (44/52) from the new onset patients and pre-diabetic relatives reacted with at least two fragments and 60% (31/52) with three or more fragments. In contrast, only one sample of the ICA512 'non-diabetes associated' sera reacted with multiple fragments (P< 10(-4)). Our findings suggest that diabetes associated anti-ICA512 autoantibodies react with multiple ICA512 epitopes while non-diabetes associated ICA512 autoantibodies may usually represent reactivity of antibodies with determinants of ICA512 unrelated to type 1 diabetes. The ability to distinguish diabetes associated from non-diabetes associated anti-ICA512 autoantibodies should provide prognostic information and more importantly suggests that even with highly specific radioassays positivity may occur unrelated to type 1 diabetes. PMID- 11908953 TI - A new middle Miocene pliopithecid from Sant Quirze, northern Spain. PMID- 11908954 TI - Oligocene sivaladapid primate from the Bugti Hills (Balochistan, Pakistan) bridges the gap between Eocene and Miocene adapiform communities in Southern Asia. AB - A new species of Guangxilemur (Sivaladapidae, Adapiformes) is described from the early Oligocene Chitarwata Formation (Bugti Member) of the Bugti Hills, Sulaiman geological Province, Balochistan, Pakistan. Guangxilemur singsilai n. sp. provides further diagnostic morphological characters from its newly described upper and lower dentitions, confirming its intermediate phylogenetic position between Eocene and Miocene Asian sivaladapid adapiforms. G. singsilai possesses moderately developed shearing and puncturing molar features and maintains lingual cusps on upper molars as in Eocene hoanghoniines; in contrast, it possesses a typical molariform P(4) as in Miocene sivaladapines. The important paleogeographic changes that have affected South Asia during the Tertiary (related to the collision between the Indian and Eurasian Plates) have played a critical role in reforming circulation and climatic differentiation. The presence in Pakistan of an unique and well-diversified Oligocene primate fauna, clearly demonstrates that South Asia maintained favourable environmental conditions during the middle Caenozoic global climatic deterioration that coincides with drastic changes in faunal structure on the whole Holarctic Province, including the extinction of adapiform primates. PMID- 11908955 TI - Morphology and affinities of new hominin cranial remains from Member 4 of the Sterkfontein Formation, Gauteng Province, South Africa. AB - Descriptions are provided of 27 hominin cranial specimens recovered from Member 4 of the Sterkfontein Formation between 1968 and 1994. Provisional statements of taxonomic affinity are given. The principal conclusion of this overview is that the bulk of the cranial remains from Member 4 are attributable to Australopithecus africanus or are consistent with the anatomy of that species, while some others are indeterminate. No specimen can be assigned confidently to a known species other than A. africanus. However, two specimens document the possibility that a second, possibly new species is represented among the Member 4 hominins, although such a species is difficult to characterize on cranial evidence alone. PMID- 11908957 TI - Faunal change, environmental variability and late Pliocene hominin evolution. AB - Global change during the late Pliocene was manifested in declining temperatures, increased amplitude of climate cycles, and shifts in the periodicity of orbital climate forcing. Linking these changes to the evolution of African continental faunas and to hominin evolution requires well-documented fossil evidence that can be examined through substantial periods of time. The Omo sequence of southern Ethiopia provides such a database, and we use it to analyze change in the abundances of mammal taxa at different levels of temporal and taxonomic resolution between 4 and 2 Ma. This study provides new evidence for shifts through time in the ecological dominance of suids, cercopithecids, and bovids, and for a trend from more forested to more open woodland habitats. Superimposed on these long-term trends are two episodes of faunal change, one involving a marked shift in the abundances of different taxa at about 2.8+/-0.1 Ma, and the second the transition at 2.5 Ma from a 200-ka interval of faunal stability to marked variability over intervals of about 100 ka. The first appearance of Homo, the earliest artefacts, and the extinction of non-robust Australopithecus in the Omo sequence coincide in time with the beginning of this period of high variability. We conclude that climate change caused significant shifts in vegetation in the Omo paleo-ecosystem and is a plausible explanation for the gradual ecological change from forest to open woodland between 3.4 and 2.0 Ma, the faunal shift at 2.8 +/-0.1 Ma, and the change in the tempo of faunal variability of 2.5 Ma. Climate forcing in the late Pliocene is more clearly indicated by population shifts within the Omo mammal community than by marked turnover at the species level. PMID- 11908956 TI - Sexual dimorphism in the Atapuerca-SH hominids: the evidence from the mandibles. AB - The pattern of sexual dimorphism in 15 mandibles from the Atapuerca-SH Middle Pleistocene site, attributed to Homo heidelbergensis, is explored. Two modern human samples of known sex are used as a baseline for establishing sexing criteria. The mandible was divided for analysis into seven study regions and differential expression of sexual dimorphism in these regions is analysed. A total of 40 continuous and 32 discrete variables were scored on the mandibles. The means method given in Regh & Leigh (Am. J. phys. Anthrop.110, 95-104, 1999) was followed for evaluating the potential of correct sex attribution for each variable. On average, the mandibles from the Atapuerca-SH site present a degree of sexual dimorphism about eight points higher than in H. sapiens samples. However, mandibular anatomy of the European Middle Pleistocene hominid records sexual dimorphism differentially. Different areas of the Atapuerca-SH mandibles exhibit quite distinct degrees of sexual dimorphism. For instance, variables of the alveolar arcade present very low or practically no sexual dimorphism. Variables related to overall size of the mandible and symphysis region present a medium degree of sex differences. Finally, ramus height, and gonion and coronoid process present a high degree of sexual dimorphism (indexes of sexual dimorphism are all above 130%). Whether this marked sexual dimorphism in specific anatomical systems affects sexual differences in body size is not completely clear and further studies are needed. Sexual differences detected in the mandible of modern humans have at least two components: differences related to musculo-skeletal development and differences related to a different growth trajectory in males and females (relative development of some of the basal border features). The Atapuerca-SH mandibles display little variation in the basal border, however. The limited variation of this mandibular region may indicate that the pattern of sexual variation in H. heidelbergensis is different enough to that of H. sapiens to caution against simple extrapolation of criteria from one pattern to the other. PMID- 11908959 TI - (+)-7S-Hydroxyxestospongin A from the marine sponge Xestospongia sp. and absolute configuration of (+)-xestospongin D. AB - The structure of the title compound, (+)-7S-hydroxyxestospongin A was solved by single-crystal X-ray diffraction analysis and the absolute stereochemistry obtained by analysis of the derived R and S Mosher's esters. The absolute configuration of xestospongin D was determined for the first time by analysis of anomalous scattering from the X-ray crystal diffraction data set. Xestospongins A, C, and D, araguspongine C, and demethylxestospongin B exhibited modest antifungal activity (MIC 30-100 g/mL) against various fluconazole-resistant Candida spp., but 7S-hydroxyxestospongin A was inactive. PMID- 11908960 TI - Cytotoxic constituents of the stem bark of Neolitsea acuminatissima. AB - Three new eudesmanolide sesquiterpenes, neolitacumone A-C (1-3), and one new benzylisoquinoline alkaloid, neolitacumonine (5), along with 27 known compounds were isolated from the stem bark of Neolitsea acuminatissima. The structures of compounds 1-3 and 5 were established on the basis of spectral and chemical evidence. Compounds 2, 3, and 20 were selectively inhibitory to Hep 2,2,15 cells with IC50 values in the range 0.24-0.04 microg/mL. Compound 20 was marginally cytotoxic to Hep G2 cells. PMID- 11908961 TI - Cyclotheonamide E4 and E5, new potent tryptase inhibitors from an Ircinia species of sponge. AB - Tryptase is a protease released from mast cells and is believed to contribute to the inflammatory process in allergic diseases including asthma. In the course of screening to find tryptase inhibitors, we isolated two new tryptase inhibitors, cyclotheonamide E4 (3) and E5 (4), from a marine sponge of the genus Ircinia. The structures of these molecules were determined by interpretation of 1H and 13C NMR spectra, and they were shown to be closely related to the previously reported cyclotheonamides E (1), E2, and E3 (2). These molecules contain two unusual amino acids, vinylogous tyrosine and alpha-ketohomoarginine, which are involved in strong activities against serine proteases. Cyclotheonamide E4 showed potent inhibitory activity against human tryptase (IC50 5.1 nM). Therefore, cyclotheonamide E4 may be useful as a therapeutic agent in the treatment of allergic diseases including asthma. PMID- 11908962 TI - New phenolic constituents from Smilax bracteata. AB - From the methanol extract of Smilax bracteata rhizomes, six new phenolic compounds, (2S,3S)-5-O-beta-D-glucopyranosyloxy-6-methyl-3'-methoxy-3,7,3' trihydroxyflavan (1), (2S,3S)-5-O-beta-D-glucopyranosyloxy-6-methyl-4'-methoxy 3,7,4'-trihydroxyflavan (2), 3beta-(3',5'-dihydroxyphenyl)-2alpha-(4' ' hydroxyphenyl)dihydrobenzofuran-5-carbaldehyde (3), (1-p-O-coumaroyl-6-O feruroyl)-beta-D-fructofuranosyl-alpha-D-glucopyranoside (4), (1-p-O-coumaroyl 3,6-di-O-feruroyl)-beta-D-fructofuranosyl-alpha-D-glucopyranoside (5), and (6-O feruroyl)-beta-D-fructofuranosyl-(6-O-acetyl)-alpha-D-glucopyranoside (6) were isolated together with five known compounds. Their structures were established by spectral data interpretation. PMID- 11908963 TI - Chemical constituents of Calophyllum brasiliensis: structure elucidation of seven new xanthones and their cancer chemopreventive activity. AB - The first study of chemical constituents of the stem bark of Calophyllum brasilienses collected in Brazil has led to the isolation and identification of seven new xanthones named brasixanthones A (1), B (4), C (5), D (6), E (2), F (3), and G (10), together with 10 known xanthones. Among the xanthones isolated in this study, 4, 5, 6, and 11 were found to exhibit significant inhibitory activity against 12-O-tetradecanoylphorbol-13-acetate induced Epstein-Barr virus early antigen activation in Raji cells. PMID- 11908964 TI - Microbial transformations of isosteviol. AB - Microbial transformations of the tetracyclic diterpenoid isosteviol (ent-16 ketobeyeran-19-oic acid) (2) have revealed that isosteviol is metabolized by Cunninghamella bainieri, Actinoplanes sp., Mucor recurvatus, and Cunninghamella blakesleeana to yield five new metabolites, ent-11alpha,12alpha-dihydroxy-16 ketobeyeran-19-oic acid (5), ent-11alpha,12alpha,17-trihydroxy-16-ketobeyeran-19 oic acid (6), ent-12alpha,15alpha-dihydroxy-16-ketobeyeran-19-oic acid (7), ent 7alpha,15alpha-dihydroxy-16- ketobeyeran-19-oic acid (8), and ent-9alpha-hydroxy 16-ketobeyeran-19-oic acid (9), together with three known metabolites, ent-7alpha hydroxy-16-ketobeyeran-19-oic acid (3), ent-7beta-hydroxy-16-ketobeyeran-19-oic acid (4), and ent-12alpha-hydroxy-16-ketobeyeran-19-oic acid (10). The structures of these metabolites were established on the basis of HRFABMS and 1D and 2D NMR spectral data. In addition, metabolites 3-10 were tested for antihypertensive activity and were found to be less active than the parent compound 2. PMID- 11908965 TI - Microbial transformations of two lupane-type triterpenes and anti-tumor-promoting effects of the transformation products. AB - Microbial transformation of betulin (1), a lupane-type triterpene obtained from the bark extract of white birch (Betula platyphylla Sukatshev var. japonica), and its chemical oxidation product, betulonic acid (2), by the fungus Chaetomium longirostre yielded 4,28-dihydroxy-3,4-seco-lup-20(29)-en-3-oic acid (3) and 4 hydroxy-3,4-seco-lup-20(29)-ene-3,28-dioic acid (4) from 1, and 4,7beta,17 trihydroxy-3,4-seco-28-norlup-20(29)-en-3-oic acid (5) and 7 beta,15 alpha dihydoxy-3-oxolup-20(29)-en-28-oic acid (6) from 2. The four metabolites, 3-6, along with 1 and 2, were evaluated for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O tetradecanoylphorbol-13-acetate (TPA) in Raji cells as a primary screening test for inhibitors of tumor promotion. All of the triterpenes tested showed potent inhibitory effects, with the four metabolites 3-6 exhibiting the more potent effects. PMID- 11908966 TI - New diarylheptanoids and diarylheptanoid glucosides from the rhizomes of Tacca chantrieri and their cytotoxic activity. AB - Two new diarylheptanoids (1, 2) and seven new diarylheptanoid glucosides (3-9) were isolated from the rhizomes of Tacca chantrieri. Their structures were determined by spectroscopic analysis, including 2D NMR data, and the results of hydrolytic cleavage. The absolute configurations of the 3,5-dihydroxyheptane moieties of the new diarylheptanoids were determined to be 3R and 5R by the application of the CD exciton chirality method to the corresponding 3,5-bis-p bromobenzoyl derivatives. The cytotoxic activities of the isolated compounds and some derivatives against HL-60 human promyelocytic leukemia cells, HSC-2 human oral squamous carcinoma cells, and normal human gingival fibroblasts (HGF) are reported. PMID- 11908967 TI - Benzoylphloroglucinol derivatives from Hypericum scabrum. AB - Nine new polyprenylated benzoylphloroglucinol derivatives, hyperibones A-I (1-9), were isolated from the aerial parts of the Uzbekistan medicinal plant Hypericum scabrum. Their structures were determined mainly on the basis of spectroscopic evidence (2D NMR and HRMS). Compounds 1, 2, and 4 showed mild in vitro antibacterial activity against methicillin-resistance Staphylococus aureus (MRSA) and methicillin-sensitive Staphylococus aureus (MSSA). PMID- 11908968 TI - Further new staurosporine derivatives from the ascidian Eudistoma toealensis and its predatory flatworm Pseudoceros sp. AB - Three new indolocarbazole alkaloids, 3-hydroxy-4'-N-methylstaurosporine (3), 3 hydroxy-4'-N-demethylstaurosporine (4), and 3'-demethoxy-3'-hydroxy-4'-N demethylstaurosporine (5), were isolated from the marine ascidian Eudistoma toealensis and its predatory flatworm Pseudoceros sp. in addition to two known staurosporines. The structures were determined by 1D and 2D homonuclear and 1H detected heteronuclear NMR spectroscopy and from comparisons with published data. CD measurements for these five staurosporine derivatives, as well as the previously described seven staurosporines, are reported, confirming that all derivatives possess the 2'S,3'R,4'R,6'R configuration. PMID- 11908969 TI - Cytotoxic prenylated xanthones and the unusual compounds anthraquinobenzophenones from Cratoxylum sumatranum. AB - Six new xanthones, cratoxyarborenones A-F (1-6), were isolated from the leaves, twigs, and/or stem bark of Cratoxylum sumatranum along with the known compound, vismione B (9), as active constituents by bioassay-directed fractionation using the KB human cancer cell line cytotoxicity assay. In addition, two novel anthraquinobenzophenones, cratoxyarborequinones A (7) and B (8), and two known compounds, 2,4,6-trihydroxybenzophenone 4-O-geranyl ether and delta-tocotrienol, were obtained as inactive constituents. PMID- 11908970 TI - Rare sesquiterpenes from the algicolous fungus Drechslera dematioidea. AB - From the inner tissue of the marine red alga Liagora viscida the fungus Drechslera dematioidea was isolated. After mass cultivation, the fungus was investigated for its secondary metabolite content, and 10 new sesquiterpenoids [isosativenetriol (1), drechslerines A (2) and B (3), 9-hydroxyhelminthosporol (5), drechslerines C-G (6-10), and sativene epoxide (12)] were isolated. Compounds 8 and 10 exhibited antiplasmodial activity against Plasmodium falciparum strains K1 and NF54. The known compounds helminthosporol (4), cis sativenediol (11), isocochlioquinone A (14), isocochlioquinone C (15), and cochlioquinone B (16) were also isolated. All structures were elucidated using spectroscopic methods, mainly 1D and 2D NMR and MS. PMID- 11908971 TI - New erythrolides from the Caribbean gorgonian octocoral Erythropodium caribaeorum. AB - Seven new briarane diterpenes, erythrolides K-Q (1-7), as well as the known compounds erythrolides A, B, C, F, and J have been isolated from samples of the Caribbean gorgonian Erythropodium caribaeorumcollected at Buccoo Reef and Flying Reef, Tobago. The structures of the new compounds were determined by high resolution 1H and 13C NMR spectroscopy utilizing COSY, HMBC, HMQC, and NOESY experiments. The structures of erythrolides K and P were confirmed and their relative stereochemistry determined by X-ray crystallographic analysis. PMID- 11908972 TI - Polyphenolic glycosides and oligosaccharide multiesters from the roots of Polygala dalmaisiana. AB - Four new polyphenolic glycosides, dalmaisiones A-D (1-4), 16 new oligosaccharide multiesters, dalmaisioses A-P (5, 7-21), and one known tetrasaccharide multiester, reiniose G (6), were isolated from the roots of Polygala dalmaisiana. The structures of the new compounds were elucidated on the basis of chemical and spectroscopic evidence. PMID- 11908973 TI - New prenylated bromoquinols from the green alga Cymopolia barbata. AB - Six new prenylated bromohydroquinones, 3'-methoxy-7-hydroxycymopol (1), 3 hydroxycymopolone (2), 3,7-dihydroxycymopolone (3), 7-hydroxycymopochromanone (4), 7-hydroxycymopochromenol (5), and a related 6-hydroxy derivative of cymopochromenol (6), have been isolated from the green marine alga Cymopolia barbata. The structures of these cymopol-related metabolites were determined by spectral methods. PMID- 11908974 TI - Oligomeric acylphloroglucinols from myrtle (Myrtus communis). AB - The dimeric nonprenylated acylphloroglucinol semimyrtucommulone (6) was obtained from the leaves of myrtle (Myrtus communis) as a 2:1 mixture of two rotamers. The known trimeric phloroglucinol myrtucommulone A (1) was also isolated and characterized spectroscopically as a silylated cyclized derivative (5). Myrtucommulone A showed significant antibacterial activity against multidrug resistant (MDR) clinically relevant bacteria, while semimyrtucommulone was less active. PMID- 11908975 TI - Dimeric and trimeric hydrolyzable tannins from Quercus coccifera and Quercus suber. AB - Three new hydrolyzable tannins, cocciferins D(1) (1), D(2) (2), and T(1) (4), were isolated from the leaves of Quercus coccifera. Cocciferin D(2) (2) and two additional new tannins, cocciferins D(3) (3) and T(2) (5), were also obtained from the leaves of Quercus suber. Their oligomeric structures were elucidated on the basis of spectroscopic methods and chemical evidence. Compounds 2, 3, and 5 were rare oligomers possessing glucose cores with both open-chain and pyranose forms. PMID- 11908976 TI - Dammarane-type triterpene saponins from Panax japonicus. AB - Six new dammarane-type saponins (1-6), together with 11 known saponins (7-17), were isolated from Ye-Sanchi, the underground part of Panax japonicus collected in the South of Yunnan Province, China. Their structures were elucidated by chemical and spectroscopic means. PMID- 11908977 TI - Flavonoid glycosides from Rhazya orientalis. AB - Six new flavonoid glycosides, quercetin 3-O-alpha-L-rhamnopyranosyl(1-->6)-[alpha L-rhamnopyranosyl(1-->2)]-(4-O-trans-p-coumaroyl)-beta-D-galactopyranoside-7-O alpha-L-rhamnopyranoside (1), quercetin 3-O-alpha-L-rhamnopyranosyl(1-->6)-[alpha L-rhamnopyranosyl(1-->2)]-(3-O-trans-p-coumaroyl)-beta-D-galactopyranoside-7-O alpha-L-rhamnopyranoside (2), isorhamnetin 3-O-alpha-L-rhamnopyranosyl(1-->6) [alpha-L-rhamnopyranosyl(1-->2)]-(4-O-trans-p-coumaroyl)-beta-D-galactopyranoside 7-O-alpha-L-rhamnopyranoside (3), isorhamnetin 3-O-alpha-L-rhamnopyranosyl(1-->6) [alpha-L-rhamnopyranosyl(1-->2)]-(3-O-trans-p-coumaroyl)-beta-D-galactopyranoside 7-O-alpha-L-rhamnopyranoside (4), isorhamnetin 3-O-alpha-L-rhamnopyranosyl(1-->6) [alpha-L-rhamnopyranosyl(1-->2)]-(4-O-cis-p-coumaroyl)-beta-D-galactopyranoside-7 O-alpha-L-rhamnopyranoside (5), and isorhamnetin 3-O-alpha-L-rhamnopyranosyl(1- >6)-[alpha-L-rhamnopyranosyl(1-->2)]-(4-O-trans-feruloyl)-beta-D galactopyranoside-7-O-alpha-L-rhamnopyranoside (6), were isolated from the dried aerial parts of Rhazya orientalis. The structures of 1-6 were determined by spectroscopic and chemical means. PMID- 11908978 TI - Phoriospongin A and B: two new nematocidal depsipeptides from the Australian marine sponges Phoriospongia sp. and Callyspongia bilamellata. AB - Bioassay-directed fractionation of two southern Australian sponges, Phoriospongia sp. and Callyspongia bilamellata, yielded two new nematocidal depsipeptides, identified as phoriospongins A (1) and B (2). The structures of the phoriospongins were determined by detailed spectroscopic analysis and comparison with the previously reported sponge depsipeptide cyclolithistide A (3), as well as ESIMS and HPLC analysis of acid hydrolysates. It is noteworthy that the unique and yet structurally related metabolites 1-3 are found in sponges spanning three taxonomic orders, Poescilosclerida, Haplosclerida, and Lithistida. PMID- 11908980 TI - Equilibrating isomers: bromoindoles and a seco-xanthine encountered during a study of nematocides from the southern Australian marine sponge Hymeniacidon sp. AB - Bioassay-directed fractionation of a Hymeniacidon sp. yielded as nematocidal agents the equilibrating E/Z bromoindole ethyl esters 1 and 2 and corresponding methyl esters 3 and 4. Also isolated for the first time as a natural product was an equilibrating mixture of seco-xanthine formamides, attributed the trivial name hymeniacidin (5). The structure for 5 was assigned on the basis of detailed spectroscopic analysis and total synthesis. PMID- 11908979 TI - Bioactive metabolites from a marine-derived strain of the fungus Emericella variecolor. AB - From a marine-derived strain of the fungus Emericella variecolor, varitriol (1), varioxirane (2), dihydroterrein (3), and varixanthone (4), besides the known mold metabolites ergosterol, terrein, shamixanthone, and tajixanthone hydrate, were identified. The chemical structures of 1-4 were established by means of spectroscopic techniques and some chemical transformations. In the NCI's 60-cell panel, varitriol (1) displayed increased potency toward selected renal, CNS, and breast cancer cell lines. Varixanthone (4) showed antimicrobial activity. PMID- 11908981 TI - Dantaxusins C and D, two novel taxoids from Taxus yunnanensis. AB - Two new taxane diterpenes, dantaxusin C (1) and dantaxusin D (2), were isolated from an ethanol extract of the aerial parts of Taxus yunnanensis along with 14 known taxoids. All structures were established on the basis of 1D and 2D NMR and HREIMS spectroscopic methods. PMID- 11908982 TI - Renealtins A and B, new diarylheptanoids with a tetrahydrofuran ring from the seeds of Renealmia exaltata. AB - Two new diarylheptanoids, renealtins A (1) and B (2), have been isolated from the seeds of the Brazilian medicinal plant Renealmia exaltata ("Pacova-catinga", Zingiberaceae), and their structures were elucidated by spectroscopic data. Renealtins A (1) and B (2) are the first example of naturally occurring diarylheptanoids containing a tetrahydrofuran ring. PMID- 11908983 TI - Tunichrome sp-1: new pentapeptide tunichrome from the hemocytes of Styela plicata. AB - A modified pentapeptide has been isolated from the hemocytes of the ascidian Styela plicata. The structure of the peptide was determined by Edman sequence analysis, mass spectrometry, and NMR spectroscopy with the stereochemistry assigned by acid hydrolysis followed by both (a) GC-MS of the volatile amino acid derivatives on a chiral column and (b) ultrasensitive detection of fluorescent diasteromeric derivatives of the component amino acids after reversed-phase HPLC. The peptide L-DOPA-L-DOPA-Gly-L-Pro-dcdeltaDOPA (where DOPA = 3,4 dihydroxyphenylalanine and dcdeltaDOPA = decarboxy-(E)-alpha,beta-dehydro-3,4 dihydroxyphenylalanine) we designate as tunichrome Sp-1. PMID- 11908984 TI - Potent cytotoxic lignans from Justicia procumbens and their effects on nitric oxide and tumor necrosis factor-alpha production in mouse macrophages. AB - A new lignan glycoside, 4-O-alpha-L-arabinopyranosyl-(1' "-->2' ')-beta-D apiofuranosyldiphyllin (2), named procumbenoside A, and 11 known compounds were isolated from the whole plant of Justicia procumbens. The structure of 2 was established by spectral analysis and chemical methods. The known compounds justicidin A (1), diphyllin (3), and tuberculatin (4) showed potent cytotoxic effects against a number of cancer cells in vitro. Compounds 1 and 4 also strongly enhanced tumor-necrosis factor-alpha (TNF-alpha) generation from mouse macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS). PMID- 11908985 TI - ent-Kaurane and beyerane diterpenoids from Excoecaria agallocha. AB - The roots of Excoecaria agallocha yielded four new diterpenoids, ent-3 beta,20 epoxy-3 alpha,6 alpha-dihydroxykaur-16-ene (agallochin F) (1), 3beta,20-epoxy-3 alpha-hydroxybeyer-15-ene (agallochin G) (2), 3 beta,20:15R,16S-diepoxy-3 alpha beyeranol (agallochin H) (3), and 3 beta,20-epoxy-3 alpha,6 alpha-dihydroxy-18 nor-beyer-15-ene (agallochin I) (4), along with three known derivatives, 2 acetoxy-1,15-beyeradiene-3,12-dione (5), 2-hydroxy-1,15-beyeradiene-3,12-dione (6), and ent-kauran-16 beta-ol-3-one. The structures of 1-4 were determined by spectroscopic (NMR and MS) data interpretation. PMID- 11908986 TI - Clavosolides A and B, dimeric macrolides from the Philippines sponge Myriastra clavosa. AB - A specimen of the sponge Myriastra clavosa from the Philippines contained two dimeric macrolides, clavosolides A and B. Clavosolide A is a symmetrical dimer, while clavosolide B is rendered unsymmetrical by the replacement of one of the methoxyl groups by a hydroxyl group. The structures of the clavosolides were elucidated by interpretation of spectroscopic data. PMID- 11908987 TI - New flavonol-phenylbutadiene adducts from the leaves of Alpinia flabellata. AB - Two new flavonol-phenylbutadiene adducts, rel-5-hydroxy-7,4'-dimethoxy-2' 'S (2,4,5-trimethoxy-E-styryl)tetrahydrofuro[4' 'R,5' 'R:2,3]flavanonol (1) and rel 5-hydroxy-7,4'-dimethoxy-3' 'S-(2,4,5-trimethoxy-E-styryl)tetrahydrofuro[4' 'R,5' 'R:2,3]flavanonol (2), were isolated from the leaves of Alpinia flabellata, along with three known compounds, 2,4,5-trimethoxybenzoic acid, 2,4,5 trimethoxycinnamic acid, and 5-hydroxy-3,7,4'-trimethoxyflavone. The structures of 1 and 2 were determined by spectroscopic interpretation. PMID- 11908988 TI - Two new ring A-rearranged clerodane diterpenes, dunniana acids A and B, from Clausena dunniana. AB - Two ring A-rearranged clerodane diterpenes named dunniana acids A (1) and B (2) were isolated from the aerial parts of Clausena dunniana. The structures of 1 and 2 were determined using spectral methods. PMID- 11908989 TI - Halogenated metabolites from the new Okinawan red alga Laurencia yonaguniensis. AB - A novel brominated diterpene based on the rare neoirieane skeleton, named neoirietetraol (1), has been isolated along with a halogenated C15 acetogenin, (3Z)-laurenyne (2), from a new Laurencia species, L.yonaguniensis Masuda et Abe, species inedita, collected at Yonaguni Island, Okinawa Prefecture, Japan. The structures of these metabolites were elucidated by spectroscopic data (IR, 1H NMR, 13C NMR, 2D NMR, and MS). Neoirietetraol (1) was toxic to the brine shrimp (Altemia salina; LC50, 40.1 microM) and also showed weak antibacterial activities against two marine bacteria, Alcaligenes aquamarinus and Escherichia coli. PMID- 11908990 TI - Phomadecalins A-D and phomapentenone A: new bioactive metabolites from Phoma sp. NRRL 25697, a fungal colonist of Hypoxylon stromata. AB - Five new natural products, phomadecalins A-D (1-4) and phomapentenone A (5), have been obtained from cultures of Phoma sp. (NRRL 25697), a fungal colonist isolated from the stromata of Hypoxylon sp. The structures of these compounds were elucidated through a series of 1D and 2D NMR experiments. Compounds 1-4 display activity against Gram-positive bacteria. PMID- 11908991 TI - Prenylated isoflavonoids from the root bark of Erythrina vogelii. AB - Four new prenylated isoflavonoids, vogelins D-G (1-4), were isolated from the CH2Cl2 extract of Erythrina vogelii root bark in addition to the known compounds isolupalbigenin (5), ficuisoflavone (6), ulexone (7), isochandalon (8), and isoderrone (9). The structures 1-4 were elucidated by spectroscopic and chemical methods. The absolute configuration of compound 1 was determined on the basis of its CD spectrum. Possible biogenetic relationships among the E. vogelii isoflavonoids are briefly discussed. PMID- 11908992 TI - Renieramide, a cyclic tripeptide from the Vanuatu sponge Reniera n. sp. AB - The polar extract of the Vanuatu sponge Reniera n. sp., which showed immunomodulating activity in preliminary tests, was found to contain a cyclic tripeptide, which we named renieramide (1). This metabolite is identical to a synthetic derivative mentioned in a patent concerning the preparation of cyclic peptides of the OF4949 family of anticancer agents. We describe here the first isolation of this metabolite from natural sources and its complete characterization by spectroscopic and chemical approaches. Renieramide (1) possesses a 17-membered cyclic side-chain-linked biphenyl ether skeleton, typical of the class that includes the natural products OF4949 I-IV, K13, and eurypamides. A tridimensional model of 1, obtained by NMR restrained molecular mechanics and dynamics, is also presented. PMID- 11908993 TI - Nobiloside, a new neuraminidase inhibitory triterpenoidal saponin from the marine sponge Erylus nobilis. AB - A neuraminidase inhibitor, nobiloside (1), was isolated from the marine sponge Erylus nobilis Thiele, 1903. Its structure was determined as a penasterol trisaccharide. The absolute configurations were determined by NMR and chiral GC analysis. It inhibited neuraminidase from the bacterium Clostridium perfringens with an IC50 value of 0.46 microg/mL. PMID- 11908994 TI - Agastinol and agastenol, novel lignans from Agastache rugosa and their evaluation in an apoptosis inhibition assay. AB - Investigation of the whole plant of Agastache rugosa resulted in the isolation of two new lignan compounds. Their structures were elucidated as (8S,7'R,8'S)-4 hydroxybenzoic acid 4-(4-hydroxy-3-methoxybenzyl)-2-(4-hydroxy-3 methoxyphenyl)tetrahydrofuran-3-ylmethyl ester (agastinol, 1) and (7'R,8'S)-4 hydroxybenzoic acid 4-(hydroxy-3-methoxybenzylidene)-2-(4-hydroxy-3 methoxyphenyl)-tetrahydrofuran-3-ylmethyl ester (agastenol, 2). Agastinol and agastenol inhibited etoposide-induced apoptosis in U937 cells with IC50 values of 15.2 and 11.4 microg/mL, respectively. PMID- 11908995 TI - New lanostanoids from the fungus Ganoderma concinna. AB - Three new compounds, 5 alpha-lanosta-7,9(11),24-triene-3beta-hydroxy-26-al (1), 5 alpha-lanosta-7,9(11),24-triene-15 alpha-26-dihydroxy-3-one (2), and 8 alpha,9 alpha-epoxy-4,4,14 alpha-trimethyl-3,7,11,15,20-pentaoxo-5 alpha-pregnane (3), were isolated from Ganoderma concinna along with 12 known compounds. The structures of compounds 1 and 2 were determined on the basis of MS and NMR studies. The structure of 3 was determined by MS, NMR, and single-crystal X-ray diffraction. Compounds 1, 2, and 3 induce apoptosis in human promyelocytic leukemia HL-60 cells, as indicated by examining the morphological features of cells and detection of DNA fragmentation by gel electrophoresis. PMID- 11908996 TI - Cloning and characterization of the bleomycin biosynthetic gene cluster from Streptomyces verticillus ATCC15003. AB - Bleomycin (BLM) biosynthesis has been studied as a model for hybrid peptide polyketide natural product biosynthesis. Cloning, sequencing, and biochemical characterization of the blm biosynthetic gene cluster from Streptomyces verticillus ATCC15003 revealed that (1) the BLM hybrid peptide-polyketide aglycon is assembled by the BLM megasynthetase that consists of both nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) modules; (2) BlmIX/BlmVIII/BlmVII constitute a natural hybrid NRPS/PKS/NRPS system, serving as a model for both hybrid NRPS/PKS and PKS/NRPS systems; (3) the catalytic sites appear to be conserved in both hybrid NRPS/PKS and nonhybrid NRPS or PKS systems, with the exception of the KS domains in the hybrid NRPS/PKS systems that are unique; (4) specific interpolypeptide linkers may play a critical role in intermodular communication to facilitate the transfer of the growing intermediates between the interacting NRPS and/or PKS modules; (5) post-translational modification of the BLM megasynthetase has been accomplished by a single PPTase with broad carrier protein specificity; and (6) BlmIV/BlmIII-templated assembly of the BLM bithiazole moiety requires intriguing protein juxtaposition and modular recognition. These results lay the foundation to investigate the molecular basis for intermodular communication between NRPS and PKS in hybrid peptide-polyketide natural product biosynthesis and set the stage for engineering novel BLM analogues by genetic manipulation of genes governing BLM biosynthesis. PMID- 11908997 TI - Improvements in gene therapy: averting the immune response to adenoviral vectors. AB - Gene therapy is an interesting approach for the correction of defective genes, the treatment of cancer and the introduction of immunomodulatory genes. Various techniques for gene transfer into cells or tissues have been developed within the last decade; these can be divided generally into viral and nonviral gene transfer systems. Nonviral techniques include the liposome- or gene gun-mediated introduction of therapeutic genes; however, the efficiency of gene transfer by these applications is still very low. In contrast, viruses have optimised their strategies for efficient infection of virtually any cell type in a mammalian organism. The genetic modification of genomes from different virus families (Adenoviridae, Retroviridae, Herpesviridae) led to the development of gene therapy vectors with a similar capacity to infect cells or tissues as that of wild type viruses. In contrast to wild type viruses, gene therapy vectors are engineered to transfer therapeutic genes into the target cells or tissues. In addition, they have lost their capacity for replication in target cells, because of the removal of essential genes, which allows replication only in specialised packaging cell lines engineered for the production of recombinant viruses. Despite considerable progress over the past decade in the generation of gene transfer systems with reduced immunogenic properties, the remaining immunogenicity of many gene therapy vectors is still the major hurdle, preventing their frequent application in clinical trials. Recombinant adenoviruses have been shown to be promising vectors for gene therapy, since they are able to transduce both quiescent and proliferating cells very efficiently. However, a major disadvantage of adenoviral vectors lies in the activation of both the innate and adaptive parts of the recipient's immune system when applied in vivo. The inflammatory responses induced by adenovirus particles can be very strong and can be fatal in patients treated with these adenoviral constructs. Therefore, many experiments have been performed in the effort to prevent these inflammatory responses mediated by adenoviral particles. The depletion of cell populations responsible for these inflammatory responses as well as the application of immunosuppressive drugs have been investigated. Moreover, the generation of less immunogenic adenoviral vectors by further genetic modification within the adenoviral genome has led to vectors with reduced immunogenic properties. Both strategies to reduce inflammatory responses against adenoviral particles are discussed in this review. PMID- 11908998 TI - Genetic polymorphisms influencing therapy and susceptibility to rejection in organ allograft recipients. AB - Solid organ transplantation during the past 30 years has developed from an experimental procedure into routine clinical practice. The current repertoire of immunosuppressive agents has made a major contribution to transplant survival; however, problems in different areas still need to be overcome. Several gene polymorphisms are supposed to influence immunosuppressive therapy and susceptibility to rejection. Therefore, a priority of transplant biologists is to estimate individual patient risk and to characterise the genetic profile of patients in need of a transplant in order to optimise the use of a scarce resource such as organs from cadaver donors, and to avoid serious drug-induced adverse effects. Polymorphisms in genes encoding tumour necrosis factor-alpha (TNFalpha), interleukin (IL)-6, IL-10, interferon-gamma (IFNgamma), transforming growth factor-beta (TGFbeta) and thiopurine S-methyltransferase (TPMT) can have significant effects on an individual's risk of rejection, as well as their ability to tolerate immunosuppressive therapy. Genotyping of known polymorphisms in these genes may in the future contribute to our ability to individualise immunosuppressive therapy in organ transplant recipients. PMID- 11908999 TI - Chlamydia vaccines: strategies and status. AB - The ultimate goal of current chlamydial vaccine efforts is to utilise either conventional or modern vaccinology approaches to produce a suitable immunisation regimen capable of inducing a sterilising, long-lived heterotypic protective immunity at mucosal sites of infection to curb the severe morbidity and worldwide prevalence of chlamydial infections. This lofty goal poses tremendous challenges that include the need to clearly define the relevant effectors mediating immunity, the antigens responsible for inducing these effectors, the anti chlamydial action(s) of effectors, and establishment of the most effective method of vaccine delivery. Tackling these challenges is further compounded by the biological complexity of chlamydia, the existence of multiple serovariants, the capacity to induce both protective and deleterious immune effectors, and the occurrence of asymptomatic and persistent infections. Thus, novel molecular, immunological and genetic approaches are urgently needed to extend the frontiers of current knowledge, and develop new paradigms to guide the production of an effective vaccine regimen. Progress made in the last 15 years has culminated in various paradigm shifts in the approaches to designing chlamydial vaccines. The dawn of the current immunological paradigm for antichlamydial vaccine design has its antecedence in the recognition that chlamydial immunity is mediated primarily by a T helper type1 (Th1) response, requiring the induction and recruitment of specific T cells into the mucosal microenvironment. Additionally, the ancillary role of humoral immune response in complementing the Th1-driven protective immunity, through ensuring adequate memory and optimal Th1 response during a reinfection, has been recognised. With continued progress in chlamydial genomics and proteomics, select chlamydial proteins, including structural, membrane and secretory proteins, are being targeted as potential subunit vaccine candidates. However, the development of an effective adjuvant, delivery vehicle or system for a potential subunit vaccine is still an elusive objective in these efforts. Promising delivery vehicles include DNA and virus vectors, bacterial ghosts and dendritic cells. Finally, a vaccine still represents the best approach to protect the greatest number of people against the ocular, pulmonary and genital diseases caused by chlamydial infections. Therefore, considering the urgency and the enormity of these challenges, a partially protective vaccine preventing certain severe sequelae would constitute an acceptable short-term goal to control Chlamydia. However, more research efforts and support are needed to achieve the worthy goal of protecting a significant number of the world's population from the devastating consequences of chlamydial invasion of the human mucosal epithelia. PMID- 11909000 TI - Prolonging organ allograft survival: potential role of nitric oxide scavengers. AB - A growing number of studies suggest a key role of nitric oxide (NO) derived from the inducible NO synthase (iNOS) isoform as a signalling molecule leading to acute organ transplant rejection. Current theory suggests that NO targets certain tissue proteins for nitrosylation or nitration leading to inhibition of enzyme/protein function and to cell death via apoptosis. Gene expression of iNOS and formation of nitrotyrosine residues have been confirmed in biopsies of rejecting grafts in humans. Experimental attempts to delay graft rejection by treatment with iNOS enzyme inhibitors have yielded conflicting results. An alternative strategy to alter rejection mediated by NO is to scavenge and/or neutralise the actions of excess NO, thereby by-passing the inhibition of iNOS enzyme activity. This review summarises recent laboratory evidence that new experimental NO scavengers/neutralisers have potential value to prolong graft survival. To date, various metal-based NO scavenging/neutralising compounds have been shown to enhance cardiac allograft survival in the absence of immunosuppression. When used in combination with low-dose cyclosporin, these agents produce a synergistic action to enhance graft survival or even to produce "permanent graft survival" under certain prolonged drug regimens. A portion of this benefit may be accounted for by the property of some of these compounds to display immunosuppressant and anti-inflammatory activity in vivo. These properties are based on findings including the following: (i) attenuating cell infiltration into the graft; (ii) attenuating activation of NFkappaB (a transcription factor important for upregulation of various inflammatory genes); (iii) attenuating cyclin D3 gene expression (a marker of cell proliferation; (iv) antagonising autoimmune activation (as determined by attenuated cytokine gene expression in splenocytes isolated from treated animals but stimulated for several days ex vivo in mixed lymphocyte cultures). PMID- 11909001 TI - Mechanisms of intravenous immunoglobulin action in the treatment of autoimmune disorders. AB - Intravenous immunoglobulins (IVIg) are therapeutic preparations of normal human immunoglobulin (Ig) G obtained from pools of blood from more than 1000 healthy donors, and exert immunomodulatory effects in autoantibody-mediated and T-cell mediated autoimmune disorders and systemic inflammatory diseases. IVIg mechanisms of action in autoimmune diseases have been extensively analysed during the last 15 years and include the following: (i) interaction of the IgG Fc fragment with Fc receptors on leucocytes and endothelial cells; (ii) interaction of infused IgG with complement proteins; (iii) monocyte and lymphocyte modulation of synthesis and release of cytokines and cytokine antagonists; (iv) modulation of cell proliferation and reparation; (v) neutralisation of circulating autoantibodies; (vi) selection of immune repertoires; and (vii) interaction with other cell surface molecules on T and B lymphocytes. PMID- 11909002 TI - Bacteriophages: potential treatment for bacterial infections. AB - Bacteriophages (phages) are viruses of bacteria that can kill and lyse the bacteria they infect. After their discovery early in the 20th century, phages were widely used to treat various bacterial diseases in people and animals. After this enthusiastic beginning to phage therapy, problems with inappropriate use and uncontrolled studies and ultimately the development of antibacterials caused a cessation of phage therapy research in the West. However, a few institutions in Eastern Europe continued to study and use phages as therapeutic agents for human infections. The alarming rise in antibacterial resistance among bacteria has led to a review of the Eastern European studies and to the initiation of controlled experiments in animal models. These recent studies have confirmed that phages can be highly effective in treating many different types of bacterial infections. The lethality and specificity of phages for particular bacteria, the ability of phages to replicate within infected animal hosts, and the safety of phages make them efficacious antibacterial agents. Although there are still several hurdles to be overcome, it appears likely that phage therapy will regain a role in both medical and veterinary treatment of infectious diseases, especially in the scenario of emerging antibacterial resistance. PMID- 11909004 TI - Cancer vaccine--Antigenics. AB - Antigenics is developing a therapeutic cancer vaccine based on heat-shock proteins (HSPs). The vaccine [HSPPC-96, Oncophage] is in a pivotal phase III clinical trial for renal cancer at 80 clinical sites worldwide. The trial is enrolling at least 500 patients who are randomised to receive surgical removal of the primary tumour followed by out-patient treatment with Oncophage((R)) or surgery only. This study was initiated on the basis of results from a pilot phase I/II study and preliminary results from a phase II study in patients with renal cell cancer. In October 2001, Oncophage was designated as a fast-track product by the Food and Drug Administration in the US for the treatment of renal cell carcinoma. Oncophage is in phase I/II trials in Italy for colorectal cancer (30 patients) and melanoma. The trials in Italy are being conducted at the Istituto dei Tumouri, Milan (in association with Sigma-Tau). Preliminary data from the phase II trial for melanoma was presented at the AACR-NCI-EORTC International Conference in Florida, USA, in October 2001. Oncophage is also in a phase I/II (42 patients) and a phase II trial (84 patients) in the US for renal cell cancer, a phase II trial in the US for non-Hodgkin's lymphoma (35 patients), a phase II trial in the US for sarcoma (20-35 patients), a phase I/II trial in the US for melanoma (36 patients), and phase I/II trials in Germany for gastric (30 patients) and pancreatic cancers. A pilot phase I trial in patients with pancreatic cancer began in the US in 1997 with 5 patients enrolled. In November 2000, Antigenics announced that this trial had been expanded to a phase I/II study which would now include survival as an endpoint and would enroll 5 additional patients. The US trials are being performed at Memorial Sloan Kettering Cancer Center and the M.D. Anderson Cancer Center. The trials in Germany are being carried out at Johannes Gutenberg-University Hospital, Mainz. Oncophage is an autologous vaccine consisting of purified complexes of tumour derived HSPs linked to tumour antigen peptides. When these HSPPC are readministered to a patient following surgery or biopsy of the tumour, the antigenic tumour peptides are expressed on the surface of potent antigen presenting cells of the immune system, such as macrophages and dendritic cells. This stimulates a much more powerful anti-tumour immune response than that generated by expression of the same antigens by the tumour cell. Thus, Antigenics autologous HSP technology is attractive because it is highly specific for individual patients and circumvents the need for identification of specific antigens for individual cancers (i.e. it does not require definition of the antigenic epitopes on cancer cells) and it overcomes the immune tolerance associated with various tumours. Oncophage is manufactured in a 10-hour process from surgically resected autologous tumour. A minimum of 1-3g of tumour tissue is required to produce enough Oncophage for a course of treatment. The major limiting factor for producing Oncophage from a particular cancer is the ability to purify HSP from that cancer. From clinical studies to date, Antigenics has been able to produce HSP from 100, 98, 90, 71 and 30% of colorectal carcinoma, renal cell carcinoma, melanoma, gastric cancer and pancreatic cancer tumours, respectively. The low success rate with pancreatic cancers is because of the high concentration of proteases in that tissue type. HSPs are a family of highly conserved proteins present in the cells of all organisms. They function as molecular chaperones, assisting the correct folding of polypeptides and aiding intracellular protein transport. In addition, HSPs associate with a broad range of peptides derived from intracellular protein degradation, including antigenic peptides produced in tumour cells. Antigenics has exclusively licensed worldwide rights to its HSP immunotherapeutic complexes from Mount Sinai School of Medicine and Fordham University in the USA. On 3 November 1998, Antigenics was issued a US patent (5,830,464) covering immunotherapy in which antigen-presenting cells are isolated and mixed with heat shock protein-antigen complexes purified from patients' tumours. The patent was issued to Fordham University, New York, US, who subsequently licensed it to Antigenics. Antigenics has an agreement with Sigma Tau, under the terms of which the latter company will fund 2 clinical trials in return for an option to market Oncophage in Italy, Portugal, Spain and Switzerland. Antigenics also has an agreement with Medison for marketing of Oncophage in Israel. PMID- 11909005 TI - FGF-4 gene therapy GENERX--Collateral Therapeutics. AB - Collateral Therapeutics and Schering AG in Germany are developing a gene therapy product, GENERX for coronary artery disease. Based on the terms of the agreement, Schering or its affliates will be responsible for conducting and financing phase II/III clinical trials which are currently underway in the US and Europe. In particular, Berlex Labs (the US subsidiary of Schering AG), is involved in developing the gene therapy in the US. GENERX is an angiogenic gene therapy which triggers the production of a protein that stimulates new blood vessel growth providing an alternative route for blood to bypass clogged and blocked arteries in the heart. GENERX involves a one-time, non-surgical delivery of an adenovirus vector containing the human fibroblast growth factor-4 (FGF-4) into coronary arteries via a standard catheter. The FGF-4 gene was licensed from New York University. Collateral Therapeutics has been granted a US patent for "gene transfer-mediated angiogenesis therapy" for the nonsurgical administration of angiogenic genes for coronary and peripheral vascular disease. The patented technology has been licensed from the University of California. Collateral and Berlex have initiated pivotal phase IIb/III trials with GENERX in the US and Europe. The US-based study will evaluate the safety and efficacy of GENERX in patients with stable exertional angina due to coronary artery disease. The European-based study will evaluate patients with advanced coronary artery disease who are not considered candidates for interventions such as angioplasty and bypass surgery and/or patients who are unlikely to have positive outcomes from such interventions. Both studies, of a multicentre, randomised, double-blind and placebo-controlled design, will evaluate 2 dose levels of GENERX which will be non-surgically administered to the heart via intracoronary infusion through a standard cardiac catheter. Collateral also plans to develop a non-surgical gene therapy product using the FGF-4 gene for the treatment of patients with heart failure. In a blinded placebo-controlled study in a pig model of pacing-induced heart failure, intracoronary delivery of human FGF-4 expressed in an adenovirus vector showed significant improvement in regional cardiac function and a reduction in the size of the heart over a 3-week study period. If these results translated favourably to humans, FGF-4 gene therapy may be a therapeutic option for patients with dilated heart failure. Collateral Therapeutics has also announced a research collaboration with Targeted Genetics on the use of viral vectors to deliver therapeutic genes in cardiovascular disease. Under the terms of the agreement, Targeted Genetics and Collateral Therapeutics each have the option to collaborate further to use Targeted Genetics' recombinant adeno associated viral vector to treat congestive heart failure. In such an event, Targeted Genetics would be responsible for constructing and manufacturing the vector, and Collateral Therapeutics will fund the costs of future collaboration. Either party may terminate this agreement at any time upon 30 days prior written notice. PMID- 11909006 TI - Management of children with severe asthma exacerbation in the emergency department. AB - Although acute asthma is a very common cause of emergency department visits in children, there is as yet insufficient evidence for the establishment of a standardized treatment protocol. The aim of this review is to describe updated information on the management of asthma exacerbations in the pediatric emergency department. Oxygen is the first-line treatment of acute asthma exacerbations in the emergency department to control hypoxemia. It is accompanied by the administration of beta(2)-adrenoceptor agonists followed by corticosteroids. beta(2)-Adrenoceptor agonists have traditionally been administered by nebulization, although spacers have recently been introduced and proven, in many cases, to be as effective as nebulization. Oral prednisolone, with its reliability, simplicity, convenience and low cost, should remain the treatment of choice for the most severe asthma exacerbations, when the lung airways are extremely contracted and filled with secretions. Recently, several studies have shown that high-dose inhaled corticosteroids are at least as effective as oral corticosteroids in controlling moderate to severe asthma attacks in children and therefore should be considered an alternative treatment to oral corticosteroids in moderate to severe asthma attacks. Studies of other drugs have shown that ipratropium bromide may be given only in addition to beta(2)-adrenoceptor agonists; theophylline has no additional benefit, and magnesium sulfate has no clear advantage. Comprehensive asthma management should also include asthma education, measures to prevent asthma triggers, and training in the use of inhalers and spacers. Proper management will avoid most asthma attacks and reduce admission and readmission to emergency departments. PMID- 11909007 TI - State of care for hemophilia in pediatric patients. AB - On demand treatment of bleeding episodes is still the main approach to hemophilia care of patients of any age. Prompt infusion of coagulation factor concentrate in the home setting allows treatment of hemorrhages at early onset, reducing the incidence of complications and improving the quality of life. Nevertheless, the technological evolution and progressive improvement in the safety of therapeutic products have changed the management of the disease, particularly in children. The current availability of safer concentrates has drastically reduced the risk of transmission of blood-borne infections. Innovative approaches, such as early primary prophylaxis and immune tolerance induction, have become feasible and their introduction represents a major advance in the achievement of the main therapeutic goals: control of the bleeding diathesis and elimination of inhibitors. Prophylactic regimens have been shown to be effective in preventing the occurrence not only of joint bleeding but also of arthropathy when started early in children with severe hemophilia. Inhibitor development still represents the main complication of hemophilia treatment, making concentrate administration ineffective. Immune tolerance induction by daily infusion of coagulation factor concentrate was shown to eradicate the inhibitors in 63 to 83% of patients. These intensive treatment regimens are administered at home to very young children and create the problem of adequate venous access. Subcutaneous venous ports have been used in patients with hemophilia if peripheral veins could not be frequently accessed; however, the risk of infection is an important limit to their use. PMID- 11909008 TI - Post-traumatic stress disorder in children and adolescents: epidemiology, diagnosis and treatment options. AB - Post-traumatic stress disorder (PTSD) is a common psychiatric condition in childhood and adolescence. Rates vary widely depending upon the type of trauma exposure. Interpersonal traumas, such as rape or physical abuse, are more likely to result in PTSD than exposure to natural or technological disaster. Clinical presentations are exceedingly complex and children with PTSD are at increased risk of having comorbid psychiatric diagnoses. Because of its complexity and frequent occurrence with other disorders, assessment of PTSD necessitates a broad based evaluation utilizing multiple informations and structured instruments specific to the symptoms of PTSD in youth. Cognitive-behavioral therapy (CBT) is the treatment of first choice. Pharmacological agents for PTSD treatment have received little empirical investigation in childhood. Pharmacological treatment is used to target disabling symptoms of the disorder, which limit psychotherapy or life functioning, by helping children to tolerate working through distressful material in therapy and life. Pharmacological treatment should be based on a stepwise approach utilizing broad spectrum medications such as the selective serotonin reuptake inhibitors as first-line agents. Comorbid conditions should be identified and treated with appropriate medication or psychosocial interventions. Treatment algorithms are provided to guide rational medication strategies for children and adolescents with PTSD, subsyndromal PTSD, and in PTSD that is comorbid with other psychiatric conditions of childhood. Reduction in even one debilitating symptom of PTSD can improve a child's overall functioning across multiple domains. PMID- 11909009 TI - Diagnosis and management of pediatric myocarditis. AB - Myocarditis is an insidious inflammatory disorder of the myocardium. As a clinical entity, it has been recognized for two centuries, but it defies traditional diagnostic tests. A greater understanding of the immune response underlying the pathobiology of the disorder can lead to a more rational therapeutic approach. The presentation, course and therapeutic options appear to be different in the pediatric compared with the adult population. An understanding of the difference between fulminant and acute progressive myocarditis has led to successful treatment strategies. A variety of new therapies are available, including antiviral agents, immunosuppression, and modulation of the biological response to inflammation. The specific question for patients with myocarditis is whether regimens designed to reduce or eliminate inflammation can provide clinical benefits compared with conventional heart failure therapy. This review highlights pathological mechanisms, modalities of diagnosis, and novel therapies which may improve outcomes. PMID- 11909010 TI - Management of uveitis in pediatric patients: special considerations. AB - Uveitis refers to inflammation involving the uvea or middle coat of the eye. This condition occurs uncommonly, particularly in persons aged approximately N2nu, with nu=0.75+/-0.01. This provides an experimental test of the theoretical value nu=3 / 4 of the critical exponent for a self avoiding random walk in two dimensions. The measured probability distribution P(r) is compared with the universal function of the scaling theory. PMID- 11909046 TI - Urn model of separation of sand. AB - We introduce an urn model that describes spatial separation of sand. In this dynamical model, in a certain range of parameters spontaneous symmetry breaking takes place and equipartitioning of sand into two compartments is broken. The steady-state equation for an order parameter, a critical line, and the tricritical point on the phase diagram are found exactly. The master equation and the first-passage problem for the model are solved numerically and the results are used to locate first-order transitions. Exponential divergence of a certain characteristic time shows that the model can also exhibit very strong metastability. In certain cases characteristic time diverges as N(z), where N is the number of balls and z=1 / 2 (critical line), 2 / 3 (tricritical point), or 1 / 3 (limits of stability). PMID- 11909047 TI - Ratchet-induced segregation and transport of nonspherical grains. AB - We consider through simulations the behavior of elongated grains on a vibrating ratchet-shaped base. We observe differences in layer velocity profile and in net grain velocity for grains that are composed of one, two, or three collinear spheres. In the case of mixtures of different species of grains, we demonstrate layer-by-layer variation in the average velocity as well as layer segregation of species, and show that horizontal separation of the species can be achieved using this geometry. We also find that the addition of a small number of shorter grains to a sample of long grains provides a lubrication effect that increases the velocity of the long grains. PMID- 11909048 TI - Interface view of directed sandpile dynamics. AB - We present a directed unloading sand-box-type avalanche model, driven by slowly lowering the retaining wall at the bottom of the slope. The avalanche propagation in the two-dimensional surface is related to the space-time configurations in one dimensional Kardar-Parisi-Zhang (KPZ) interface growth. We relate the scaling exponents of the avalanche cluster distribution to those for the growing surface. The numerical results are close but deviate significantly from the exact KPZ values. This might reflect stronger than usual corrections to scaling or be more fundamental, due to correlations between subsequent space-time interface configurations. PMID- 11909049 TI - Basin of attraction of a bounded self-organized critical state. AB - The robustness of the self-organized critical (SOC) state observed in the motion of an annular plate rotating over a granular medium is studied in this paper. In particular, we investigate the effect of parameters to which the emergent SOC state may be sensitive, including the initialization scheme, driving velocity, and confining pressure. The results indicate that the critical state is not a universal attractor, but has a finite basin of attraction. Furthermore, this state is only one of the three observed, which compare well with subcritical, critical, and supercritical states. The results call into question the precise definition of the term "self-organized criticality," an issue we address. PMID- 11909050 TI - Finite width of shear zones. AB - We present some experimental and numerical results based on a simple model designed to give an estimate for the width of a shear zone. We conclude that the observed finite size of the shear zone can be associated with the propagation of the force lines inside the medium. The model, based on a simple argument on the force distribution and dilatancy, predicts a width of about ten grain diameters. PMID- 11909051 TI - Orientational structure of dipolar hard-spherical colloids. AB - We have studied the orientational structure of a dipolar hard-spherical colloid on a homogeneous isotropic phase. The results are expressed as a function of the dipolar strength mu and volume fraction phi of dipolar colloids, and the refractive index of the scattering medium, n(s). The study is based on the self correlation of the orientation density of the dipolar colloids, which is the static orientational structure factor [F(q)], where q is the wave vector. The importance of this quantity is that for very low phi values, it can be probed in a depolarized light scattering experiment. We have found that the structure of the suspension is better observed for high n(s). F(q) presents a different behavior for dilute and dense concentrations, it is also observed that the position of its minimum depends on phi. The response of a dipolar colloid due to its collective orientational behavior is also studied, using as an "ordering parameter" the static orientational structure factor at q=0[F(q=0)]. The study is performed for isochores as a function of mu. We have divided the analysis into five regimes, from very low to very high phi; values, i.e., phi=0.005 24, 0.1, 0.2, 0.35, and 0.45. Our analysis suggests that the dipolar colloid evolves to an orientationally ordered phase when the dipolar strength is increased, for all concentrations except for the lowest value case, phi=0.005 24. When phi=0.1 the dipolar colloid reaches the transition suddenly, whereas for the very low regime, the slope of F(q=0) first increases as if the dipolar colloid would evolve to an orientationally ordered phase; but near the transition the slope is inverted, resulting in a no global orientational order. Thus, our results suggest that in the very low regime a dipolar colloid may have a reentrant transition. PMID- 11909052 TI - Ordering and short-time orientational diffusion in dipolar hard-spherical colloids. AB - Orientational hydrodynamic functions and short-time, self-orientational and collective orientational diffusion coefficients of dipolar hard-spherical colloids are performed on a homogeneous isotropic phase, as functions of the wave vector q, for various values of the volume fraction and the dipolar strength of the macroparticles. The calculation is based on the dynamic orientational structure factor, which is the time-dependent self-correlation of the orientation density. We assume that the time evolution of the orientation density is given by the Smoluchoswki's equation, taking into account the hydrodynamic interactions as well as the dipolar interaction. The former are considered assuming pairwise additivity. The importance of the dynamic orientational structure factor is that its initial slope can be measured in a depolarized light scattering experiment. The results predict a different behavior for dilute and for dense dipolar colloids. The ordering phenomena are studied via the ordering coefficients, which are the orientational hydrodynamic functions at q=0. The results show that as the dipolar colloid evolves to the instability line, the translational ordering velocity increases while the rotational one reduces. The short-time orientational diffusion coefficients at q=0 are also performed. They predict that near to the instability line, the dipolar colloid diffuses translationally more than rotationally. At very dilute concentration the dipolar colloid presents an unexpected dynamical behavior, which seems to indicate that the colloid could be evolving to a reentrant phase. PMID- 11909053 TI - Anisotropy of the structure factor of magnetic fluids under a field probed by small-angle neutron scattering. AB - Small-angle neutron scattering is used to measure the two-dimensional diffraction pattern of a monophasic magnetic colloid, under an applied magnetic field. This dipolar system presents in zero field a fluidlike structure. It is well characterized by an interaction parameter K(0)(T) proportional to the second virial coefficient, which is here positive, expressing a repulsion of characteristic length kappa-10. Under the field a strong anisotropy is observed at the lowest q vectors. The length kappa-10 remains isotropic, but the interaction parameter K(T) becomes anisotropic due to the long-range dipolar interaction. However, the system remains stable, the interaction being repulsive in all directions. Thus we do not observe any chaining of the nanoparticles under magnetic field. On the contrary, the revealed structure of our anisotropic colloid is a lowering of the concentration fluctuations along the field while the fluidlike structure, observed without field, is roughly preserved perpendicularly to the field. It expresses a strong anisotropy of the Brownian motion of the nanoparticles in the solution under applied field. PMID- 11909054 TI - Binding of biological effectors on magnetic nanoparticles measured by a magnetically induced transient birefringence experiment. AB - We have investigated the relaxation of the magnetically induced birefringence in a suspension of magnetic nanoparticles in order to detect the binding reaction of polyclonal antibodies on the particle surface. The birefringence relaxation is driven by the rotational diffusion of the complex formed by the magnetic nanoparticles bound to the antibody and thus is directly related to the hydrodynamic size of this complex. Birefringence relaxations are well described by stretched exponential laws revealing a polydisperse distribution of hydrodynamic diameters. Comparing the size distribution of samples with different initial ratios of immunoglobuline added per magnetic nanoparticles, we evidence the graft of an antibody on particle and eventually the onset of particles aggregation. Measurements on samples separated in size by gel filtration demonstrate the robustness of our experiment for the determination of size distribution and its modification due to the adsorption of a macromolecule. The immunoglobuline binding assay is performed comparatively for ionic magnetic nanoparticles with different coatings. PMID- 11909055 TI - Constant bond breakup probability model for reversible aggregation processes. AB - Reversible aggregation processes were simulated for systems of freely diffusing sticky particles. Reversibility was introduced by allowing that all bonds in the system may break with a given probability per time interval. In order to describe the kinetics of such aggregation-fragmentation processes, a fragmentation kernel was developed and then used together with the Brownian aggregation kernel for solving the corresponding kinetic master equation. The deduced fragmentation kernel considers a single characteristic lifetime for all bonds and accounts for the cluster morphology by averaging over all possible configurations for clusters of a given size. It became evident that the simulated cluster-size distributions could be described only when an additional fragmentation effectiveness was considered. Doing so, the stochastic solutions were in good agreement with the simulated data. PMID- 11909056 TI - Phase transition analogous to Bose-Einstein condensation in systems of noninteracting surfactant aggregates. AB - Ideal bosons and a classical system of monomers that aggregate forming noninteracting ring polymers are known to have the same partition function. So, the ring polymers have a phase transition, the analogue of Bose-Einstein condensation of bosons. At this phase transition macroscopic polymers are formed. The link between these systems is made via Feynman's path integrals: these integrals are the same for the trajectories of the bosons in imaginary time and for the configurations of the polymers. We show that a transition of this general form occurs within a whole class of aggregating systems. Examples are the lamellae formation in suspensions of disclike micelles or the emulsification failure observed in water-oil-surfactant emulsions. As with bosons, the transition occurs even when aggregates do not interact. The lambda-transition in 4He is believed to be Bose-Einstein condensation modified by interatomic interactions. We suggest that interaggregate interactions too only modify the transition we have found. PMID- 11909057 TI - Phase equilibria and glass transition in colloidal systems with short-ranged attractive interactions: application to protein crystallization. AB - We have studied a model of a complex fluid consisting of particles interacting through a hard-core and short-range attractive potential of both Yukawa and square-well form. Using a hybrid method, including a self-consistent and quite accurate approximation for the liquid integral equation in the case of the Yukawa fluid, perturbation theory to evaluate the crystal free energies, and mode coupling theory of the glass transition, we determine both the equilibrium phase diagram of the system and the lines of equilibrium between the supercooled fluid and the glass phases. For these potentials, we study the phase diagrams for different values of the potential range, the ratio of the range of the interaction to the diameter of the repulsive core being the main control parameter. Our arguments are relevant to a variety of systems, from dense colloidal systems with depletion forces, through particle gels, nanoparticle aggregation, and globular protein crystallization. PMID- 11909058 TI - Thermodiffusion in magnetic colloids evidenced and studied by forced Rayleigh scattering experiments. AB - This paper shows how forced Rayleigh scattering can be used as an experimental tool for studying thermodiffusion (Soret effect). The systems investigated are magnetic colloids of different types. A framework including thermodiffusion and dielectrophoresis is described in which the evolutions of temperature and of colloid concentration are clearly distinguished. The framework is then shown to account for experiments on steady-state concentration gratings coupled with transient temperature ones, and the parameters are determined therefrom. Dielectrophoretic forces are found to be negligible. Studying different types of magnetic colloids with various dilution rates shows that the sign of the Soret effect is controlled by the nature of the particle coating made up of electrostatic charges or of surfactant, and that its mechanism is located at the nanoparticle core-solvent interface. PMID- 11909059 TI - Kinetics of macroion coagulation induced by multivalent counterions. AB - Due to the strong correlations between multivalent counterions condensed on a macroion, the net macroion charge changes sign at some critical counterion concentration. This effect is known as the charge inversion. Near this critical concentration the macroion net charge is small. Therefore, short range attractive forces between macroions dominate Coulomb repulsion and lead to their coagulation. The kinetics of macroion coagulation in this range of counterion concentrations is studied. We calculate the Coulomb barrier between two approaching like charged macroions at a given counterion concentration. Two different macroion shapes (spherical and rodlike) are considered. A new "self regulated" regime of coagulation is found. As the size of aggregates increases, their charge and Coulomb barrier also grow and diminish the sticking probability of aggregates. This leads to a slow, logarithmic increase of the aggregate size with time. PMID- 11909060 TI - History-dependent rheology of a surfactant hexagonal phase. AB - The time-dependent response of a surfactant hexagonal phase of a sodium dodecyl sulphate/pentanol/cyclohexane/brine system to stepped strain is investigated. The dynamics of the system is found to be governed by strain- and noise-induced yielding of the domains of the system. The effects of the applied strain magnitude and the ionic strength of the brine on the character of the transitions experienced by the system are reported. PMID- 11909061 TI - Using patterned substrates to promote mixing in microchannels. AB - Using a lattice Boltzmann model for fluid dynamics, we investigate the flow and phase behavior of a binary fluid moving over a patterned substrate within a microchannel. The binary fluid consists of two immiscible components, A and B, and this liquid is subjected to a Poiseuille flow. The substrate is decorated with a checkerboard pattern of A- and B-like patches. Through a coupling of hydrodynamics and thermodynamics, each component is driven to flow from the nonwettable domains to wettable regions. As a consequence, the A and B fluids undergo extensive mixing within the microchannels. We investigate how the degree of mixing depends on the size of the patches, the velocity of the imposed flow field, and the characteristics of the fluid. The results provide guidelines for creating localized "mixing stations" within microfluidic devices. The findings also reveal how a combination of imposed flow fields and surface patterning can be exploited to control the phase behavior of complex fluids. PMID- 11909062 TI - Real-coded lattice gas model for ternary amphiphilic fluids. AB - We have developed an amphiphilic surfactant model in the framework of the real coded lattice gas (RLG), in order to analyze the dynamics and structure of ternary fluids. Formation of both the oil-in-water and the bicontinuous microemulsion phases, as well as the reduction of surface tension by adsorption of surfactant at interface are successfully reproduced in numerical simulations. Our model is simple in terms of description and implementation, however, complex structures and dynamic behavior of the ternary fluids emerge from the collective dynamics of the RLG and surfactant particles. PMID- 11909063 TI - Glasslike relaxation of labyrinthine domain patterns. AB - A spatial analysis of globally disordered (labyrinthine) stripe domain patterns in thin ferrimagnetic garnet films is applied to investigate the pattern evolution. After demagnetization of the sample we obtain a branched (fernlike) structure. By periodic modulation of the magnetic field the number of the branches diminishes and a labyrinthine pattern develops. We describe the evolution of the pattern by a measure extracted from the curvature of the border line of the magnetic domains. The relaxation of this measure is found to be nonexponential and can be described by the Kohlrausch-Williams-Watts law. PMID- 11909064 TI - Inside the hysteresis loop: multiplicity of internal states in confined fluids. AB - We study the equilibrium and stability of metastable states and capillary condensation hysteresis of a Lennard-Jones fluid in cylindrical pores by means of the canonical ensemble density functional theory and gauge cell Monte Carlo simulations. We demonstrate a possibility for the existence of multiple laterally uniform internal states of equal density inside the hysteresis loop. The region of multiple states is bounded by the states of zero compressibility. The internal states can be stabilized in Monte Carlo simulations constraining the density fluctuations. PMID- 11909065 TI - Kinetics of formation of a phase with an arbitrary stoichiometric composition in a multicomponent solid solution. AB - A kinetic theory of nucleation and growth of a evolving phase with a given stoichiometric composition in a multicomponent solid solution is developed. It is assumed naturally that the phase grows as a result of individual atom incorporation into the phase domain in a stoichiometric ratio. As it is shown, for the case of phase formation in a multicomponent system the basic kinetic equations, describing the nucleation-growth process, can be reduced formally to the respective expression derived for nucleation-growth processes in one component systems. However, the effective diffusion coefficients and the effective supersaturation are expressed as nontrivial combinations of the thermodynamic and kinetic parameters of the different components involved in the phase formation process. In the determination of these properties, the theory is not restricted in its applicability to perfect solutions but extended to phase formation in real mixtures. Thus, the theory may be applied directly towards the interpretation of experimental data. In the present paper, particular attention is devoted to the analysis of the two stages of the overall transformation process: (1) the stage of quasi-steady-state nucleation and (2) the transient stage of coarsening. As the results of this analysis, the quasi-steady-state nucleation rate, the number of clusters formed via nucleation and growth, and the time evolution of the cluster size distributions are established. Moreover, estimates are given for the duration of the different stages of the transformation process. PMID- 11909066 TI - Electrorheological fluid under elongation, compression, and shearing. AB - Electrorheological (ER) fluid based on zeolite and silicone oil under elongation, compression, and shearing was investigated at room temperature. Dc electric fields were applied on the ER fluid when elongation and compression were carried out on a self-constructed test system. The shear yield stress, presenting the macroscopic interactions of particles in the ER fluid along the direction of shearing and perpendicular to the direction of the electric field, was also obtained by a HAAKE RV20 rheometer. The tensile yield stress, presenting the macroscopic interactions of particles in the ER fluid along the direction of the electric field, was achieved as the peak value in the elongating curve with an elongating yield strain of 0.15-0.20. A shear yield angle of about 15 degrees 18.5 degrees reasonably connected tensile yield stress with shear yield stress, agreeing with the shear yield angle tested well by other researchers. The compressing tests showed that the ER fluid has a high compressive modulus under a small compressive strain lower than 0.1. The compressive stress has an exponential relationship with the compressive strain when it is higher than 0.1, and it is much higher than shear yield stress. PMID- 11909067 TI - Anisotropic capillary interactions and jamming of colloidal particles trapped at a liquid-fluid interface. AB - We determine the capillary attraction and equilibrium configurations of particles trapped at a liquid-fluid interface due to the pinning of their contact line. We calculate analytically the asymptotic interaction energy between two particles and, numerically, the multibody energy landscape for up to four contacting particles. Our results are consistent with recent experiments. We show that a system composed of a large number of such particles behaves as a jammed system. PMID- 11909068 TI - Monte Carlo simulation of sinusoidally modulated superlattice growth. AB - The fabrication of ZnSe/ZnTe superlattices grown by the process of rotating the substrate in the presence of an inhomogeneous flux distribution instead of the successively closing and opening of source shutters is studied via Monte Carlo simulations. It is found that the concentration of each compound is sinusoidally modulated along the growth direction, caused by the uneven arrival of Se and Te atoms at a given point of the sample, and by the variation of the Te/Se ratio at that point due to the rotation of the substrate. In this way we obtain a ZnSe(1 x)Tex alloy in which the composition x varies sinusoidally along the growth direction. The period of the modulation is directly controlled by the rate of the substrate rotation. The amplitude of the compositional modulation is monotonic for small angular velocities of the substrate rotation, but is itself modulated for large angular velocities. The average amplitude of the modulation pattern decreases as the angular velocity of substrate rotation increases and the measurement position approaches the center of rotation. The simulation results are in good agreement with previously published experimental measurements on superlattices fabricated in this manner. PMID- 11909069 TI - Scaling laws in adsorption on bivariate surfaces. AB - The adsorption of particles with nearest-neighbor attractive and repulsive interactions is studied through Monte Carlo simulation on bivariate surfaces characterized by patches of weak and strong adsorbing sites of size l. Patches are considered to have either a square or a strip geometry and they can be either arranged in a deterministic ordered structure or in a random way. Quantities are identified that scale obeying power laws as a function of the scale length l. The consequences of this finding are discussed for the determination of the energetic topography of the surface from adsorption measurements. PMID- 11909071 TI - Wetting transitions at soft, sliding interfaces. AB - We observe (by optical interferometry) the contact of a rubber cap squeezing a nonwetting liquid against a plate moving at velocity U. At low velocities, the contact is dry. It becomes partially wet above a threshold velocity V(c1), with two symmetrical dry patches on the rear part. Above a second velocity V(c2), the contact is totally wet. This regime U>V(c2) corresponds to the hydroplaning of a car (decelerating on a wet road). We interpret the transitions at V(c1), V(c2) in terms of a competition between (a) liquid invasion induced by shear (b) spontaneous dewetting of the liquid (between nonwettable surfaces). PMID- 11909070 TI - Evidence for tip velocity oscillations in dendritic solidification. AB - Dendritic growth experiments were conducted in the reduced-convection environment aboard the space shuttle Columbia on STS-87. Spectral analysis was performed on 30-frame/s video data during growths of isothermal dendrites. Results indicate that pivalic acid dendrites exhibit a subtle oscillatory behavior of the axial growth velocity near the tip, with a frequency component that is associated with the sidebranch formation process. PMID- 11909073 TI - Is a position-dependent stiffness relevant for the wetting phase diagram? AB - In this paper we determine the wetting phase diagram for three-dimensional systems with short-range forces assuming the presence of a position-dependent stiffness contribution as recently proposed [M.E. Fisher and A.J. Jin, Phys. Rev. Lett. 69, 792 (1992)]. We predict a discontinuous transformation of the phase diagram immediately upon moving beyond the mean-field approximation. However, in contrast to Fisher and Jin we find that a renormalization group calculation yields fluctuation-induced second-order transitions rather than fluctuation induced first-order ones. As a consequence, in all fluctuation regimes we recover the same qualitative phase diagram as predicted in the absence of a position dependent stiffness coefficient. Furthermore, recent predictions for tricritical wetting behavior remain unaffected by the stiffness contribution. PMID- 11909074 TI - Domain wall roughening in dipolar films in the presence of disorder. AB - We derive a low-energy Hamiltonian for the elastic energy of a Neel domain wall in a thin film with in-plane magnetization, where we consider the contribution of the long-range dipolar interaction beyond the quadratic approximation. We show that such a Hamiltonian is analogous to the Hamiltonian of a one-dimensional polaron in an external random potential. We use a replica variational method to compute the roughening exponent of the domain wall for the case of two dimensional dipolar interactions. PMID- 11909075 TI - Ions and nematic surface energy: beyond the exponential approximation for the electric field of ionic origin. AB - We present a general model to describe the influence of the ionic adsorption on the anisotropic part of the surface energy of a nematic liquid crystal in contact with a substrate. We show that in the limit of small adsorption energy, the exponential approximation for the electric field of ionic origin works well. In this limit, the dielectric and flexoelectric contributions to the surface energy are quadratic and linear on the density of adsorbed ions, respectively. In the opposite limit of large adsorption energy, the exponential approximation for the electric field does not work, and the two contributions to the surface energy are both found to depend linearly on the surface density of adsorbed charges. Approximated formulas reported in literature are derived from our general equations as particular cases, and their limits discussed. An expression for the surface polarization in nematic liquid crystal due to the ionic adsorption is also deduced. Our analysis is performed in the framework of the Poisson-Boltzmann theory, where dimensionless ions are treated within a mean field approach. Possible extensions of our model are indicated. PMID- 11909072 TI - Nonequilibrium self-assembly of metals on diblock copolymer templates. AB - Most studies of self-assembled systems reveal that the highest order is associated with equilibrium states of the system. By systematically studying metal decoration of diblock copolymer templates, I show that a high degree of order can arise under strongly nonequilibrium conditions. Under a wide range of conditions, thermally evaporated gold decorates ultrathin, asymmetric, polystyrene-b-polymethylmethacrylate diblock copolymer films with isolated nanoparticles. These particles aggregate into nanoparticle chains inside the polystyrene block with a selectivity approaching 100%. However, even at metal loading fractions of up to 30% by volume no coalescence into continuous nanowires is observed. This behavior is also shared by indium, tin, lead, bismuth, and silver at low coverage (<30 A nominal thickness). At high coverage (>100 A nominal thickness), however, silver self-assembles to form nanowires. One can understand the formation of the chains of nanoparticles by understanding the equilibrium state of the system (metal+polymer). The silver nanowires are highly nonequilibrium structures and, to the best of my knowledge, unexplained by existing theoretical models. Assuming an energy difference for metallic particles for either side of the diblock, a mobility difference, and an attractive interaction between metallic particles, I modeled the self-assembly of the nanowires with a Monte Carlo simulation. This Monte Carlo simulation qualitatively agrees with the formation of the silver nanowires and their relaxation to equilibrium upon moderate heating. PMID- 11909076 TI - Structural transitions in thin free-standing films of an antiferroelectric liquid crystal exhibiting the smectic-C(*)(alpha) phase in the bulk sample. AB - Optical reflectivity studies have been conducted upon thin free-standing films of an antiferroelectric liquid crystal possessing the bulk phase sequence Sm C(*)(A) Sm C(*)(alpha)-Sm A. Electrically induced field independent and field dependent structural transformations were observed above the bulk Sm C(*)(A)-Sm C(*)(alpha) transition temperature. The anisotropy of the reflectivity was measured for different states in thin films. Our data provide direct evidence of structures without the short-pitched helix in the temperature region between the antiferroelectric Sm C(*)(A) and untilted Sm A structures. PMID- 11909077 TI - Slow molecular dynamics of water in a lyotropic complex fluid studied by deuterium conventional and spin-lattice relaxometry NMR. AB - A nuclear magnetic resonance study of protons and deuterons in the mesomorphic phases of the micellar lyotropic mixture potassium laurate/1-decanol/heavy water is reported. The slow dynamical behavior of water molecules has been investigated with deuterons spin-lattice relaxation dispersion in the Larmor frequency range 10(3)cubic Ia3d phase transition was successfully detected. Through an analysis of chain-length dependence of the entropy of transition, the sequence of two cubic mesophases (with space groups Ia3d and Im3m) is deduced for thermotropic mesogens ANBC(n). It is shown that the disorder of the core arrangement decreases in the order of Sm-C-->cubic (Im3m)- >cubic (Ia3d) while that of the chain in the reverse order cubic (Ia3d)-->cubic (Im3m)-->Sm C. PMID- 11909094 TI - Electro-optic response and switchable Bragg diffraction for liquid crystals in colloid-templated materials. AB - We report optical switching studies on nematic liquid crystal incorporated into structures based on self-assembled colloids. We compare the electro-optic responses of liquid crystal imbibed into colloid-templated polymers, liquid crystal imbibed in the interstitial space of colloid crystals, and conventional polymer-dispersed liquid crystals. We characterize the Bragg diffraction of our templated liquid-crystal/polymer composites as a function of electric field and measure switching times. The response of liquid crystal in connected networks differs qualitatively from that of liquid crystal in isolated cavities. PMID- 11909095 TI - External electric-field effect on nematic anchoring energy. AB - The influence of an external field on the effective anchoring energy of a nematic liquid crystal in contact with a substrate is theoretically analyzed. Our analysis is performed on the hypothesis that the electrodes are perfectly blocking and that there is no selective ion adsorption. The proposed theory predicts an effective anchoring energy dependent on the applied dc voltage. According to the sign of the dielectric anisotropy and of the flexoelectric coefficient the dependence of the anchoring energy strength with the bias can be monotonic or not. For large bias voltage the effective anchoring energy strength tends to a constant value. Our theory is in qualitative agreement with published data investigating the influence of the bias on the saturation voltage. PMID- 11909096 TI - Area-constrained planar elastica. AB - We determine the equilibria of a rigid loop in the plane, subject to the constraints of fixed length and fixed enclosed area. Rigidity is characterized by an energy functional quadratic in the curvature of the loop. We find that the area constraint gives rise to equilibria with remarkable geometrical properties; not only can the Euler-Lagrange equation be integrated to provide a quadrature for the curvature but, in addition, the embedding itself can be expressed as a local function of the curvature. The configuration space is shown to be essentially one dimensional, with surprisingly rich structure. Distinct branches of integer-indexed equilibria exhibit self-intersections and bifurcations-a gallery of plots is provided to highlight these findings. Perturbations connecting equilibria are shown to satisfy a first-order ODE which is readily solved. We also obtain analytical expressions for the energy as a function of the area in some limiting regimes. PMID- 11909097 TI - Coexistence curve near the tricritical point in ternary polymer solutions. AB - Two-phase coexistence curves were measured for ternary solutions of bimodal polystyrene in methylcyclohexane with the molecular weight ratio near the tricritical value 23. Coexistence curves were determined by the refractive index method on a diagram of temperature versus volume fraction of total polystyrene. The diameter was strongly curved near the top. The double logarithmic plots of volume fraction difference between two coexisting phases versus reduced temperature yielded the critical exponent beta=0.250+/-0.005 for the tricritical solution and, 0.412+/-0.005 and 0.383+/-0.016 for solutions not far from the tricritical one. The former value could be compared with the classical tricritical exponent beta=1 / 4 and the latter values near 0.40 could be explained by a crossover between the ordinary critical exponent 0.32 or the tricritical exponents 1 / 4 and the classical exponent 1 / 2. PMID- 11909098 TI - Orientations of the lamellar phase of block copolymer melts under oscillatory shear flow. AB - We develop a theory to describe the reorientation phenomena in the lamellar phase of block copolymer melts under reciprocating shear flow. We show that, similar to the steady shear, the oscillating flow anisotropically suppresses fluctuations and gives rise to the [parallel]--> [perpendicular] transition. The experimentally observed high-frequency reverse transition is explained in terms of interaction between the melt and the shear-cell walls. PMID- 11909099 TI - Chain segment order in ultrathin polymer films: a deuterium NMR study. AB - The orientational order of monomerically thin films of polydimethylsiloxane melt deposited on the surfaces of cylindrical pores is investigated in this paper by solid-state deuterium nuclear magnetic resonance. Spectra demonstrate substantial orientation of the methyl groups by the surface. The quadrupolar splitting of the oriented film component saturates below one monomer layer, but decreases stepwise for thicker films. Comparisons of experimental and simulated spectra suggest a flat chain conformation in the first polymer monolayer. PMID- 11909100 TI - Salt effect on volume phase transition of a gel. AB - The salt effect on the phase transition of N-isopropylacrylamide (NIPA) gel was studied for alkali-metal chlorides (NaCl, KCl, and CsCl). Low-frequency Raman scattering experiment was conducted to know the dynamic state of water molecule under the presence of salt and its correlation to macroscopic phase behavior of the gel was investigated together with the thermodynamic activities of water molecule of aqueous alkali-metal chloride solutions. The series of swelling experiment reveal that the change in the gel volume phase transition strongly depends on the salt concentration and is related to the dehydration with respect to hydrophobic hydration. From the analysis of the reduced low-frequency Raman spectra in water and aqueous alkali-metal chlorides solutions by the use of the relaxation mode that takes into account the inertia and the non-white effects, the characteristic values of aqueous salt solutions (i.e., relaxation time and modulation speed) indicate that the addition of alkali-metal chloride to gel fluid affects the disruption of water molecules in the hydration shell around the NIPA gel and the formation of the hydrogen-bonded network structure of water around themselves, as a result of which the gel collapses. The chemical potential and the dynamic nature of water molecule at the transition points are well correlated: the chemical potentials at the transition points are almost constant whereas the structure of bulk water is changed by addition of alkali-metal chlorides or change in temperature. These results strongly suggest that the swelling ratio of N-isopropylacrylamide gel is a function of hydration degree, which is regulated by the chemical potential of water. PMID- 11909101 TI - Viscosity of entangled polystyrene thin film melts: Film thickness dependence. AB - We determined the low-shear effective viscosity of entangled polystyrene thin film melts, in the thickness range of 270, the deposit attains some static configuration, in which no deposition attempt is accepted. In 1+1 dimensions, the interface width has a limiting value W(s) approximately p(-eta), with eta=3/2, which is confirmed by numerical simulations. The dynamic scaling relation W(s)=p(-eta)f(tp(z)) is obtained in very large substrates, with z=eta. PMID- 11909126 TI - Self-consistent expansion for the molecular beam epitaxy equation. AB - Motivated by a controversy over the correct results derived from the dynamic renormalization group (DRG) analysis of the nonlinear molecular beam epitaxy (MBE) equation, a self-consistent expansion for the nonlinear MBE theory is considered. The scaling exponents are obtained for spatially correlated noise of the general form D(r-r('),t-t('))=2D(0)[r-->-r(')](2rho-d)delta(t-t(')). I find a lower critical dimension d(c)(rho)=4+2rho, above which the linear MBE solution appears. Below the lower critical dimension a rho-dependent strong-coupling solution is found. These results help to resolve the controversy over the correct exponents that describe nonlinear MBE, using a reliable method that proved itself in the past by giving reasonable results for the strong-coupling regime of the Kardar-Parisi-Zhang system (for d>1), where DRG failed to do so. PMID- 11909125 TI - Inertial effects in anomalous dielectric relaxation. AB - The inertia corrected Debye model of rotational Brownian motion of polar molecules is generalized to fractional dynamics (anomalous diffusion) in the context of the fractional Klein-Kramers equation. The fractal generalization of the Gross-Sack solution for the complex dielectric susceptibility chi(omega) for an assembly of fixed axis rotators is given. The high-frequency behavior of chi(omega) is controlled by the inertia of a dipole as in normal diffusion, so that the Gordon sum rule for dipolar absorption is satisfied ensuring a return to optical transparency at very high frequencies. PMID- 11909127 TI - Anomalous slowing down in the metastable liquid of hard spheres. AB - It is demonstrated that a straightforward extension of the Arrhenius law accurately describes diffusion in the thermodynamically stable liquid of hard spheres. A sharp negative deviation from this behavior is observed as the liquid is compressed beyond its stability limit. This dynamical anomaly can be compared with the nonlinear slowing down characteristic of the supercooled dynamics regime in liquids with continuous interaction. It is suggested that the observed dynamical transition is caused by long-time decomposition of the configuration space. This interpretation is corroborated by the observation of characteristic anomalies in the geometry of a particle trajectory in the metastable domain. PMID- 11909129 TI - Elastic coupling of silica gel dynamics in a liquid-crystal-aerosil dispersion. AB - The dynamics of a thixotropic silica aerosil gel dispersed in an octylcyanobiphenyl liquid crystal were directly probed by x-ray intensity fluctuation spectroscopy. For all samples, the time-autocorrelation function of the gel was well described by a modified-exponential function over the q range studied. Compared to a pure gel sample, a dilute (0.06 g cm(-3)) gel embedded within the liquid crystal displayed more complex and temperature dependent dynamics. Near the second-order smectic-A-to-nematic phase transition of the liquid crystal the gel relaxation became significantly more complex and slower (tau approximately 2150 s) compared to relaxations observed well within either phase. This clearly demonstrates coupling between the dynamics of the gel and the host liquid crystal, consistent with critical slowing down of smectic and director fluctuations. A random dampening field, elastically coupled to the liquid crystal, would explain the earlier observed crossover of this transition towards 3d-XY behavior. PMID- 11909128 TI - Creep rupture of viscoelastic fiber bundles. AB - We study the creep rupture of bundles of viscoelastic fibers occurring under uniaxial constant tensile loading. A fiber bundle model is introduced that combines the viscoelastic constitutive behavior and the strain controlled breaking of fibers. Analytical and numerical calculations showed that above a critical external load the deformation of the system monotonically increases in time resulting in global failure at a finite time t(f), while below the critical load the deformation tends to a constant value giving rise to an infinite lifetime. Our studies revealed that the nature of the transition between the two regimes, i.e., the behavior of t(f) at the critical load sigma(c), strongly depends on the range of load sharing: for global load sharing t(f) has a power law divergence at sigma(c) with a universal exponent of 0.5, however, for local load sharing the transition becomes abrupt: at the critical load t(f) jumps to a finite value, analogous to second- and first-order phase transitions, respectively. The acoustic response of the bundle during creep is also studied. PMID- 11909130 TI - Exact calculations of the paranematic interaction energy for colloidal dispersions in the isotropic phase of a nematogenic material. AB - In a recent paper [Phys. Rev. E 61, 2831 (2000)], Borstnik, Stark, and Zumer have studied the stability of a colloidal dispersion of micron-sized spherical particles in the isotropic phase of a nematogenic material. Close to the nematic transition, the attraction due to a surface-induced paranematic order can yield flocculation. Their calculation of the nematic-mediated interaction was based on an ansatz for the order-parameter profile. We compare it with an exact numerical calculation, showing that their results are qualitatively correct. Besides, we point out that in the considered regime, the exact interaction is extremely well approximated by a simple analytical formula which is asymptotically exact. PMID- 11909131 TI - Linearity and scaling of a statistical model for the species abundance distribution. AB - We derive a linear recursion relation for the species abundance distribution in a statistical model of ecology and demonstrate the existence of a scaling solution. PMID- 11909133 TI - Reply to "comment on 'Monte Carlo simulations for a Lotka-type model with reactant surface diffusion and interactions' ". AB - We reply to the Comment by Zhdanov [preceding paper, Phys. Rev. E 65, blacksquare, square, filled (2002)] on our recent paper [G. Zvejnieks and V. N. Kuzovkov, Phys. Rev. E 63, 051104 (2001)]. We demonstrate that our quite different viewpoints result, in fact, entirely from nonunique definitions of the master equation, which has nothing to do with neglecting important physical principles, as Zhdanov claims. PMID- 11909132 TI - Comment on "Monte Carlo simulations for a Lotka-type model with reactant diffusion and interactions". AB - Discussing the effect of adsorbate-adsorbate lateral interactions on the kinetics of heterogeneous catalytic reactions, Zvejnieks and Kuzovkov [Phys. Rev. E 63, 051104 (2001)] conclude that in the case of adsorbed particles the Metropolis Monte Carlo dynamics is meaningless and propose to use their own dynamics, which is equivalent to the Glauber dynamics. In this Comment, I show that these and other conclusions and prescriptions by Zvejnieks and Kuzovkov are not in line with the general principles of simulations of rate processes in adsorbed overlayers. PMID- 11909134 TI - Comment on "nonlinear viscosity and Grad's method". AB - In their recent paper [Phys. Rev. E 60, 4052 (1999)] Uribe and Garcia-Colin suggest that the stress tensor associated with the nonlinear viscosity formula eta=eta(0)sinh (-1)kappa/kappa (kappa=a Rayleigh dissipation function) vanishes asymptotically as the magnitude of the velocity gradient increases. In this Comment, it is pointed out that their remark is invalid, because the stress tensor asymptotically exhibits a logarithmic kappa dependence. It is also pointed out that their evolution equations for the stress tensor components are missing the terms containing the velocity gradients in the transversal directions and, as a consequence, give rise to a vanishing shear stress, contrary to the experimental evidence of gas flow in a tube. PMID- 11909136 TI - Nonuniversality of invasion percolation in two-dimensional systems. AB - Employing highly efficient algorithms for simulating invasion percolation (IP) with trapping, we obtain precise estimates for the fractal dimensions of the sample-spanning cluster, the backbone, and the minimal path in a variety of two dimensional lattices. The results indicate that these quantities are nonuniversal and vary with the coordination number Z of the lattices. In particular, while the fractal dimension D(f) of the sample-spanning cluster in lattices with low Z has the generally accepted value of about 1.82, it crosses over to the value of random percolation, D(f) approximately equal to 1.896, if Z is large enough. Since optimal paths in strongly disordered media and minimum spanning trees on random graphs are related to IP, the implication is that these problems do not also possess universal scaling properties. PMID- 11909137 TI - First-order depinning transition of a driven interface in disordered media. AB - We introduce a simple growth model which exhibits a first-order pinning-depinning (PD) transition in disordered media. In our model, a first-order PD transition is triggered by the local inertia force F(l)=pLv macro where p denotes a constant between 0 and 1, L is the system size, and v macro is the average velocity in a local region of the growing interface. If pp(c), our model shows a first-order PD transition. We measure the critical exponents characterizing the dynamical behavior of our model and explain how a first-order PD transition can occur if p>p(c). Besides the PD transitions, our model exhibits another phase transition from a fluctuating to a nonfluctuating interface with a constant velocity. PMID- 11909138 TI - Equivalence of stationary state ensembles. AB - We show that the contact process in an ensemble with conserved total particle number, as simulated recently by Tome and de Oliveira [Phys. Rev. Lett. 86, 5463 (2001)], is equivalent to the ordinary contact process, in agreement with what the authors assumed and believed. Similar conserved ensembles and equivalence proofs are easily constructed for other models. PMID- 11909135 TI - Reply to "comment on nonlinear viscosity and Grad's method ". AB - We show that while Eu's claim is true that we made a mistake regarding the asymptotic behavior of his theory is true, the correct asymptotic behavior cannot have a physical meaning. We analyze in more detail his theory for a dilute gas of rigid spheres, and show that in some cases it predicts a negative value of the xx component of the pressure tensor. PMID- 11909139 TI - Statistics of multiple sign changes in a discrete non-Markovian sequence. AB - We study analytically the statistics of multiple sign changes in a discrete non Markovian sequence psi(i)=phi(i)+ phi(i-1) (i=1,2, em leader,n) where phi(i)'s are independent and identically distributed random variables each drawn from a symmetric and continuous distribution rho(phi). We show that the probability P(m)(n) of m sign changes up to n steps is universal, i.e., independent of the distribution rho(phi). The mean and variance of the number of sign changes are computed exactly for all n>0. We show that the generating function (tilde)P(p,n)= summation operator(infinity)(m=0)P(m)(n)p(m) approximately exp[-theta(d)(p)n] for large n where the "discrete" partial survival exponent theta(d)(p) is given by a nontrivial formula, theta(d)(p)=ln[sin(-1)(square root of [1-p(2)])/square root of [1-p(2)]] for 0< or = p < or = 1. We also show that in the natural scaling limit m-->infinity, n-->infinity but keeping x=m/n fixed, P(m)(n) approximately exp[-n Phi (x)] where the large deviation function Phi(x) is computed. The implications of these results for Ising spin glasses are discussed. PMID- 11909141 TI - Multiparameter generalization of nonextensive statistical mechanics. AB - We show that the stochastic interpretation of Tsallis's thermostatistics given recently by Beck [Phys. Rev. Lett 87, 180601 (2001)] leads naturally to a multiparameter generalization. The resulting class of distributions is able to fit experimental results, which cannot be reproduced within Boltzmann's or Tsallis's formalism. PMID- 11909140 TI - Anomalous diffusion, stable processes, and generalized functions. AB - The evolution equations in real space and time corresponding to a class of anomalous diffusion processes are examined. As special cases, evolution equations corresponding to stable processes are derived using the theory of generalized functions, recovering some known results differently interpreted, and an evolution law for stable processes of order unity. PMID- 11909142 TI - Crack propagation in thin glass plates caused by high velocity impact. AB - Crack propagation within thin glass plates under high shock loading is directly observed using a high speed camera. The fractal dimension of cracks and the power law exponents of the fragment area distributions are investigated as a function of time. Two models of the fragmentation process are proposed: in one case the cracks are net-like, while in the other the cracks are tree-like, and the relations between fractal dimension and power-law exponent are estimated and compared with the experimental results. It appears that at early stages of the fragmentation process the relation is described by the latter case, while at later stages it approaches that of the former case. PMID- 11909143 TI - Epidemic dynamics in finite size scale-free networks. AB - Many real networks present a bounded scale-free behavior with a connectivity cutoff due to physical constraints or a finite network size. We study epidemic dynamics in bounded scale-free networks with soft and hard connectivity cutoffs. The finite size effects introduced by the cutoff induce an epidemic threshold that approaches zero at increasing sizes. The induced epidemic threshold is very small even at a relatively small cutoff, showing that the neglection of connectivity fluctuations in bounded scale-free networks leads to a strong overestimation of the epidemic threshold. We provide the expression for the infection prevalence and discuss its finite size corrections. The present paper shows that the highly heterogeneous nature of scale-free networks does not allow the use of homogeneous approximations even for systems of a relatively small number of nodes. PMID- 11909144 TI - Diffusion resonances in action space for an atom optics kicked rotor with decoherence. AB - We numerically investigate momentum diffusion rates for the pulse kicked rotor across the quantum to classical transition as the dynamics are made more macroscopic by increasing the total system action. For initial and late time rates we observe an enhanced diffusion peak which shifts and scales with changing kick strength, and we also observe distinctive peaks around quantum resonances. Our investigations take place in the context of a system of ultracold atoms which is coupled to its environment via spontaneous emission decoherence, and the effects should be realizable in ongoing experiments. PMID- 11909145 TI - Suppressing spatiotemporal disorder via local perturbations in an electrochemical cell. AB - We report experimental results depicting suppression of complex spatiotemporal dynamics under the influence of local periodic stimulations. In an experimental electrochemical system, applying a continuous forcing signal to one of the sites in an array of eight coupled oscillators, the naturally complex behavior of the remaining seven electrodes can be converted to periodic responses. The oscillations remain periodic as long as the forcing is active and revert back to exhibiting chaotic dynamics after the control is switched off. These results can also be interpreted as experimental realization of "phase-synchronization" induced via local driving in an extended system. A possible relevance to the experimentally observed calcium wave patterns is pointed out. PMID- 11909146 TI - High-dimensional interior crisis in the Kuramoto-Sivashinsky equation. AB - An investigation of interior crisis of high dimensions in an extended spatiotemporal system exemplified by the Kuramoto-Sivashinsky equation is reported. It is shown that unstable periodic orbits and their associated invariant manifolds in the Poincare hyperplane can effectively characterize the global bifurcation dynamics of high-dimensional systems. PMID- 11909147 TI - Reconstructing embedding spaces of coupled dynamical systems from multivariate data. AB - A method for reconstructing dimensions of subspaces for weakly coupled dynamical systems is offered. The tool is able to extrapolate the subspace dimensions from the zero coupling limit, where the division of dimensions as per the algorithm is exact. Implementation of the proposed technique to multivariate data demonstrates its effectiveness in disentangling subspace dimensionalities also in the case of emergent synchronized motions, for both numerical and experimental systems. PMID- 11909148 TI - Spatial forcing of spontaneous optical patterns. AB - A nonlinear optical system, which spontaneously forms hexagonal patterns, is exposed to a weak, spatially modulated forcing. As forcing, stationary hexagonal patterns are used under variation of their transverse wave number. In the experiment, we observe a locking when the forcing wave number is close to one of the critical wave numbers of the unforced system. Outside the locking regimes, forcing provokes spatiotemporal disorder. The system response is characterized quantitatively with respect to its dynamics and to its spatial order. PMID- 11909149 TI - Rainbow transition in chaotic scattering. AB - We study the effects of classical chaotic scattering on the differential cross section, which is the measurable quantity in most scattering experiments. We show that the fractal set of singularities in the deflection function is not, in general, reflected on the differential cross section. We show that there are systems in which, as the energy (or some other parameter) crosses a critical value, the system's differential cross-section changes from a singular function having an infinite set of rainbow singularities with structure in all scales to a smooth function with no singularities, the scattering being chaotic on both sides of the transition. We call this metamorphosis the rainbow transition. We exemplify this transition with a physically relevant class of systems. These results have important consequences for the problem of inverse scattering in chaotic systems and for the experimental observation of chaotic scattering. PMID- 11909150 TI - Effects of disorder on the wave front depinning transition in spatially discrete systems. AB - Pinning and depinning of wave fronts are ubiquitous features of spatially discrete systems describing a host of phenomena in physics, biology, etc. A large class of discrete systems is described by overdamped chains of nonlinear oscillators with nearest-neighbor coupling and subject to random external forces. The presence of weak randomness shrinks the pinning interval and it changes the critical exponent of the wave front depinning transition from 1/2 to 3/2. This effect is derived by means of a recent asymptotic theory of the depinning transition, extended to discrete drift-diffusion models of transport in semiconductor superlattices and is confirmed by numerical calculations. PMID- 11909151 TI - Ehrenfest times for classically chaotic systems. AB - We describe the quantum-mechanical spreading of a Gaussian wave packet by means of the semiclassical WKB approximation of Berry and Balazs [J. Phys. A 2, 625 (1979)]. We find that the time scale tau on which this approximation breaks down in a chaotic system is larger than the Ehrenfest times considered previously. In one dimension tau=7/6 lambda(-1)ln(A/Plancks over 2 pi), with lambda the Lyapunov exponent and A a typical classical action. PMID- 11909152 TI - Scalar variance decay in chaotic advection and Batchelor-regime turbulence. AB - The decay of the variance of a diffusive scalar in chaotic advection flow (or equivalently Batchelor-regime turbulence) is analyzed using a model in which the advection is represented by an inhomogeneous baker's map on the unit square. The variance decays exponentially at large times, with a rate that has a finite limit as the diffusivity kappa tends to zero and is determined by the action of the inhomogeneous map on the gravest Fourier modes in the scalar field. The decay rate predicted by recent theoretical work that follows scalar evolution in linear flow and then averages over all stretching histories is shown to be incorrect. The exponentially decaying scalar field is shown to have a spatial power spectrum of the form P(k) approximately k(-sigma) at wave numbers small enough for diffusion to be neglected, with sigma<1. PMID- 11909153 TI - Three-lobe-shaped equilibrium states in magnetic liquid bridges. AB - Stable 3-lobed cross sectional equilibrium shapes, invariant under rotations of 120 degrees, are reported in magnetic liquid drops confined between two horizontal plates in the presence of a vertical dc magnetic field. Their connectivity with other 4-, 3- or 2-lobed shapes as magnetic field strength varies is studied experimentally. Although 3-lobed shapes are found to evolve into bent 2-lobed ones by slowly decreasing the field strength, the reverse transition is not observed when the field strength increases. Moreover, 2-lobed shapes suffer a bending transition, by increasing the field strength, which is found to be hysteretic. PMID- 11909154 TI - Explosive energy release in magnetic shocks. AB - We show that a magnetic shock whose initial density and/or magnetic perturbation exceeds the Hugoniot limit may lead to substantial and rapid energy release in low beta plasmas (such as occur in the magnetospheres of neutron stars). We illustrate this effect for a fast Magnetohydrodynamic perturbation, as well as for large density perturbations which can be naturally created in low beta plasmas. Using the Riemann solution and simulations, we show that slow modes of finite magnitudes and Alfvenic perturbations can generate strong density perturbations. These perturbations develop into shocks, resulting in efficient energy release. PMID- 11909155 TI - Toroid moments in the momentum and angular momentum loss by a radiating arbitrary source. AB - In the context of experimental evidence concerning a nuclear toroid dipole, we briefly present here an exact but tedious calculation of the angular momentum loss, recoil force, and radiation intensity for an arbitrary source in terms of electric, magnetic, and toroid multipoles, thus emphasizing the importance of the latter in getting the results in closed forms, unbiased by approximations. Corrections to some familiar formulas from books, mostly on account of time varying toroid moments, are found and discussed. PMID- 11909156 TI - Statistical theory for incoherent light propagation in nonlinear media. AB - A statistical approach based on the Wigner transform is proposed for the description of partially incoherent optical wave dynamics in nonlinear media. An evolution equation for the Wigner transform is derived from a nonlinear Schrodinger equation with arbitrary nonlinearity. It is shown that random phase fluctuations of an incoherent plane wave lead to a Landau-like damping effect, which can stabilize the modulational instability. In the limit of the geometrical optics approximation, incoherent, localized, and stationary wave fields are shown to exist for a wide class of nonlinear media. PMID- 11909157 TI - Nematic liquid crystals: a suitable medium for self-confinement of coherent and incoherent light. AB - Nematic liquid crystals exhibit a saturable, non-instantaneous nonlinear response through light-induced reorientation. In such a material, we demonstrate that (2+1)-dimensional spatial solitary waves can be generated at milliwatt power levels not only with a coherent optical beam, but also with incoherent excitations. Self-trapping also allows the efficient guidance of a weak co polarized probe. PMID- 11909159 TI - Generalized thermodynamics of q-deformed bosons and fermions. AB - We study the thermostatistics of q-deformed bosons and fermions obeying the symmetric (q<-->q(-1)) algebra and show that it can be built on the formalism of q calculus. The entire structure of thermodynamics is preserved if ordinary derivatives are replaced by an appropriate Jackson derivative. In this framework, we derive the most important thermodynamic functions describing the q-boson and q fermion ideal gases in the thermodynamic limit. We also investigate the semiclassical limit and the low-temperature regime and demonstrate that the nature of the q deformation gives rise to pure quantum statistical effects stronger than undeformed boson and fermion particles. PMID- 11909158 TI - Rational approximation with multidimensional scattered data. AB - Accurate and efficient rational approximation schemes are presented for interpolating multidimensional scattered data with a novel weighted least-squares procedure including domain decomposition. Two particular representations of the method are formulated and the corresponding algorithms are implemented. Numerical tests on three- and six-dimensional model systems are carried out, demonstrating high efficiency and accuracy. This work was motivated by the need for multidimensional function approximation using irregular grids when solving quantum fluid dynamics equations, and the method should have broader physical applications. PMID- 11909160 TI - Criticality and oscillatory behavior in non-Markovian contact process. AB - A non-Markovian generalization of a one-dimensional contact process is being introduced in which every particle has an age and will be annihilated at its maximum age tau. There is an absorbing state phase transition which is controlled by this parameter. The model can demonstrate oscillatory behavior in its approach to the stationary state. These oscillations are also present in the mean-field approximation, which is a first-order differential equation with time delay. Studying dynamical critical exponents suggests that the model belongs to the direct percolation universality class. PMID- 11909161 TI - Exact amplitude ratio and finite-size corrections for the MxN square lattice Ising model. AB - Let f, U, and C represent, respectively, the free energy, the internal energy, and the specific heat of the critical Ising model on the MxN square lattice with periodic boundary conditions, and f(infinity) represents f for fixed M/N and N- >infinity. We find that f, U, and C can be written as N(f-f(infinity))= summation operator(infinity)(i=1)f(2i-1)/N(2i-1), U=-square root of [2]+ summation operator(infinity)(i=1)u(2i-1)/N(2i-1), and C=8 ln N/pi+ summation operator(infinity)(i=0)c(i)/N(i), i.e., Nf and U are odd functions of N(-1). We also find that u(2i-1)/c(2i-1)=1/square root of [2] and u(2i)/c(2i)=0 for 1 < or = i 1/2 the correlations appear to have no effect, and the transport process is diffusive. However, for H<1/2 and depending on the morphology of the medium, three distinct types of transport processes, namely, anomalous, Fickian, and superdiffusive transport may emerge. Moreover, if the medium is anisotropic and stratified, biased diffusion in it is characterized by power-law growth of the mean square displacements with the time in which the effective exponents characterizing the power-law oscillates log periodically with the time. This result cannot be predicted by any of the currently available continuum theories of transport in disordered media. PMID- 11909175 TI - Fracture in mode I using a conserved phase-field model. AB - We present a continuum phase-field model of crack propagation. It includes a phase-field that is proportional to the mass density and a displacement field that is governed by linear elastic theory. Generic macroscopic crack growth laws emerge naturally from this model. In contrast to classical continuum fracture mechanics simulations, our model avoids numerical front tracking. The added phase field smooths the sharp interface, enabling us to use equations of motion for the material (grounded in basic physical principles) rather than for the interface (which often are deduced from complicated theories or empirical observations). The interface dynamics thus emerges naturally. In this paper, we look at stationary solutions of the model, mode I fracture, and also discuss numerical issues. We find that the Griffith's threshold underestimates the critical value at which our system fractures due to long wavelength modes excited by the fracture process. PMID- 11909176 TI - Experimental study of the dimensionality of black-eye patterns. AB - The spatial structure of one type of Turing pattern named "black eyes" is studied in experiments using the chlorite-iodide-malonic acid reaction in spatial open reactor. The purpose of the work is to verify (or falsify) two possible theoretical interpretations of black-eye pattern formation: the projection of a body-centered-cubic (bcc) structure onto the plane vertical to the body diagonal and the spatial resonance of larger wave vectors with the fundamental ones. The latter happens when the mode of the larger wave vectors becomes linearly unstable, as the system goes far beyond the onset of Turing bifurcation. Our experimental results give evidence that black-eye patterns are not a projection of the bcc structure, so that the interpretation of a three-dimensional structure can be ruled out. The observations in the experiment also suggest another possible mechanism for the formation of black eyes. PMID- 11909177 TI - Slow dynamics and aging in a nonrandomly frustrated spin system. AB - A simple, non-disordered spin model has been studied in an effort to understand the origin of the precipitous slowing down of dynamics observed in supercooled liquids approaching the glass transition. A combination of Monte Carlo simulations and exact calculations indicates that this model exhibits an entropy vanishing transition accompanied by a rapid divergence of time scales. Measurements of various correlation functions show that the system displays a hierarchy of time scales associated with different degrees of freedom. Extended structures, arising from the frustration in the system, are identified as the source of the slow dynamics. In the simulations, the system falls out of equilibrium at a temperature T(g) higher than the entropy-vanishing transition temperature and the dynamics below T(g) exhibits aging as distinct from coarsening. The cooling rate dependence of the energy is also consistent with the usual glass formation scenario. PMID- 11909178 TI - Capacity drop due to the traverse of pedestrians. AB - In this paper, we have proposed a simplified model to describe the traffic flow when there are pedestrians traversing the road. The numerical simulation shows that the capacity of the road decreases in the presence of pedestrians. If the traffic flow rate is small, the traffic flow is basically unaffected even if some pedestrians traverse the road. However, if the flow rate exceeds a critical value, the vehicles cannot pass without delay, and a traffic jam appears. We also discuss simplified conditions of the model and accordingly present a modified model, which predicts qualitatively the same results except with a different capacity. PMID- 11909179 TI - Extended phase-space dynamics for the generalized nonextensive thermostatistics. AB - We apply a variant of the Nose thermostat to derive the Hamiltonian of a nonextensive system that is compatible with the canonical ensemble of the generalized thermostatistics of Tsallis. This microdynamical approach provides a deterministic connection between the generalized nonextensive entropy and power law behavior. For the case of a simple one-dimensional harmonic oscillator, we confirm by numerical simulation of the dynamics that the distribution of energy H follows precisely the canonical q statistics for different values of the parameter q. The approach is further tested for classical many-particle systems by means of molecular dynamics simulations. The results indicate that the intrinsic nonlinear features of the nonextensive formalism are capable of generating energy fluctuations that obey anomalous probability laws. For q<1 a broad distribution of energy is observed, while for q>1 the resulting distribution is confined to a compact support. PMID- 11909180 TI - Correlation effects in a simple model of a small-world network. AB - We analyze the effect of correlations in a simple model of a small-world network by obtaining exact analytical expressions for the distribution of shortest paths in the network. We enter correlations into a simple model with a distinguished site, by taking the random connections to this site from an Ising distribution. Our method shows how the transfer-matrix technique can be used in the new context of small-world networks. PMID- 11909181 TI - Highly clustered scale-free networks. AB - We propose a model for growing networks based on a finite memory of the nodes. The model shows stylized features of real-world networks: power-law distribution of degree, linear preferential attachment of new links, and a negative correlation between the age of a node and its link attachment rate. Notably, the degree distribution is conserved even though only the most recently grown part of the network is considered. As the network grows, the clustering reaches an asymptotic value larger than that for regular lattices of the same average connectivity and similar to the one observed in the networks of movie actors, coauthorship in science, and word synonyms. These highly clustered scale-free networks indicate that memory effects are crucial for a correct description of the dynamics of growing networks. PMID- 11909182 TI - Multifractal properties of resistor diode percolation. AB - Focusing on multifractal properties we investigate electric transport on random resistor diode networks at the phase transition between the nonpercolating and the directed percolating phase. Building on first principles such as symmetries and relevance we derive a field theoretic Hamiltonian. Based on this Hamiltonian we determine the multifractal moments of the current distribution that are governed by a family of critical exponents [psi(l)]. We calculate the family [psi(l)] to two-loop order in a diagrammatic perturbation calculation augmented by renormalization group methods. PMID- 11909183 TI - Light-induced evaporation and condensation growth of aerosol particles. AB - The nonisothermal evaporation and condensation of a particle suspended in a vapor gas mixture under the effect of resonant optical radiation is studied. A traveling light wave is absorbed by the aerosol particle and velocity selectively by vapor molecules. The dependence of the evaporation or condensation rate on optical and thermophysical properties of the particle and gaseous molecules is studied. The free-molecule regime is examined. PMID- 11909184 TI - Heterogeneous interfacial failure between two elastic blocks. AB - We investigate numerically the failure process when two elastic media, one hard and one soft that have been glued together thus forming a common interface, are pulled apart. We present three main results: (1) The area distribution of simultaneously failing glue (bursts) follows a power law consistent with the theoretically expected exponent 2.5, (2) the maximum load and displacement before catastrophic failure scale as L(2) and L(0), respectively, where L is the linear size of the system, and (3) the area distribution of failed glue regions (clusters) is a power law with exponent -1.6 when the system fails catastrophically. PMID- 11909185 TI - Extreme value statistics and traveling fronts: application to computer science. AB - We study the statistics of height and balanced height in the binary search tree problem in computer science. The search tree problem is first mapped to a fragmentation problem that is then further mapped to a modified directed polymer problem on a Cayley tree. We employ the techniques of traveling fronts to solve the polymer problem and translate back to derive exact asymptotic properties in the original search tree problem. The second mapping allows us not only to again derive the already known results for random binary trees but to obtain exact results for search trees where the entries arrive according to an arbitrary distribution, not necessarily randomly. Besides it allows us to derive the asymptotic shape of the full probability distribution of height and not just its moments. Our results are then generalized to m-ary search trees with arbitrary distribution. PMID- 11909186 TI - Duality in nonextensive statistical mechanics. AB - We revisit recent derivations of kinetic equations based on Tsallis' entropy concept. The method of kinetic functions is introduced as a standard tool for extensions of classical kinetic equations in the framework of Tsallis' statistical mechanics. Our analysis of the Boltzmann equation demonstrates a remarkable relation between thermodynamics and kinetics caused by the deformation of macroscopic observables. PMID- 11909187 TI - Influence of boundary conditions on statistical properties of ideal Bose-Einstein condensates. AB - We investigate the probability distribution that governs the number of ground state particles in a partially condensed ideal Bose gas confined to a cubic volume within the canonical ensemble. Imposing either periodic or Dirichlet boundary conditions, we derive asymptotic expressions for all its cumulants. Whereas the condensation temperature becomes independent of the boundary conditions in the large-system limit, as implied by Weyl's theorem, the fluctuation of the number of condensate particles and all higher cumulants remain sensitive to the boundary conditions even in that limit. The implications of these findings for weakly interacting Bose gases are briefly discussed. PMID- 11909188 TI - Effect of disorder on critical short-time dynamics. AB - Critical short-time dynamics in a bond-diluted Ising model is investigated in this paper using numerical simulations. The effective static and dynamic critical exponents determined by the power-law scaling are found to depend strongly on bond concentration and initial state. For weak disorder, the short-time scaling relations for the system quenched from high temperature are observed to hold. In the strong dilution limit, multiscaling relations for the system starting from the ordered state are found. Corrections to the short-time scaling are proposed. The effect of disorder on critical short-time dynamics is discussed. PMID- 11909189 TI - Anomalous scaling and Lee-Yang zeros in self-organized criticality. AB - We show that the generating functions of probability distributions in self organized criticality (SOC) models exhibit a Lee-Yang phenomenon [Phys. Rev. 87, 404 (1952)]. Namely, their zeros pinch the real axis at z=1, as the system size goes to infinity. This establishes a new link between the classical theory of critical phenomena and SOC. A scaling theory of the Lee-Yang zeros is proposed in this setting. PMID- 11909190 TI - Dynamic model for the popularity of websites. AB - In this paper, we have studied a dynamic model to explain the observed characteristics of websites in the World Wide Web. The dynamic model consists of the self-growth term for each website and the external force term acting on the website. With simulations of the model, we can explain most of the important characteristics of websites. These characteristics include a power-law distribution of the number of visitors to websites, fluctuation in the fractional growth of individual websites, and the relationship between the age and the popularity of the websites. We also investigated a few variants of the model and showed that the ingredients included in the model adequately explain the behavior of the websites. PMID- 11909191 TI - Global exponential stability of continuous-time interval neural networks. AB - This paper addresses global robust stability of a class of continuous-time interval neural networks that contain time-invariant uncertain parameters with their values being unknown but bounded in given compact sets. We first introduce the concept of diagonally constrained interval neural networks and present a necessary and sufficient condition for global exponential stability of these interval neural networks irregardless of any bounds of non-diagonal uncertain parameters in connection weight matrices. Then we extend the robust stability result to general interval neural networks by giving a sufficient condition. Simulation results illustrate the characteristics of the main results. PMID- 11909192 TI - Pseudospectral method for the Kardar-Parisi-Zhang equation. AB - We discuss a numerical scheme to solve the continuum Kardar-Parisi-Zhang equation in generic spatial dimensions. It is based on a momentum-space discretization of the continuum equation and on a pseudospectral approximation of the nonlinear term. The method is tested in (1+1) and (2+1) dimensions, where it is shown to reproduce the current most reliable estimates of the critical exponents based on restricted solid-on-solid simulations. In particular, it allows the computations of various correlation and structure functions with high degree of numerical accuracy. Some deficiencies that are common to all previously used finite difference schemes are pointed out and the usefulness of the present approach in this respect is discussed. PMID- 11909193 TI - Effect of the subsurface oxygen diffusion on the Ziff-Gulari-Barshad catalytic reaction model. AB - We study a version of the Ziff-Gulari-Barshad model where we include the diffusion of oxygen atoms between the uppermost layer and the subsurface. When a CO molecule impinges the surface, it occupies a single site, while the O2 molecule needs two neighboring sites to be adsorbed. The oxidation of the CO molecule occurs only at the top layer, and this happens whenever a CO molecule is nearest neighbor of an O atom. Through the pair mean-field approximation we determine the phase diagram of the model for different values of the diffusion rate of oxygen atoms between the subsurface and the top layer. The diagram exhibits a continuous line that separates regions displaying O-poisoned and non-O poisoned states. We show that above a critical value of the diffusion rate of oxygen atoms from the subsurface to the top layer, there is no more oxygen poisoning for any nonzero value of the diffusion rate from the top layer to the subsurface. This behavior is also verified in Monte Carlo simulations. PMID- 11909194 TI - Kink production in the presence of impurities. AB - The production of kinks during a quench in an overdamped regime of phi(4) model is investigated. Expelling defects from regions of nonzero force coming from the impurity are predicted. PMID- 11909195 TI - Numerical confirmation of late-time t(1/2) growth in three-dimensional phase ordering. AB - Results for the late-time regime of phase ordering in three dimensions are reported, based on numerical integration of the time-dependent Ginzburg-Landau equation with nonconserved order parameter at zero temperature. For very large systems (700(3)) at late times, t> or =150, the characteristic length grows as a power law, R(t) approximately t(n), with the measured n in agreement with the theoretically expected result n=1/2 to within statistical errors. In this time regime R(t) is found to be in excellent agreement with the analytical result of Ohta, Jasnow, and Kawasaki [Phys. Rev. Lett. 49, 1223 (1982)]. At early times, good agreement is found between the simulations and the linearized theory with corrections due to the lattice anisotropy. PMID- 11909196 TI - Model for glass transition in a binary fluid from a mode coupling approach. AB - We consider the mode coupling theory (MCT) of glass transition for a binary fluid. The equations of nonlinear fluctuating hydrodynamics for the compressible fluid are obtained with a proper choice of slow variables which correspond to the conservation laws in the system. The resulting model equations are used to obtain a coupled set of nonlinear integro-differential equations for the various correlations of partial density fluctuations. These equations are then solved self consistently in the long-time limit to locate the dynamic transition in the system. The transition point from our model is at considerably higher density than predicted in the other existing MCT models for binary systems. PMID- 11909197 TI - Generalized Young's equation for rough and heterogeneous substrates: a microscopic proof. AB - We derive by microscopic techniques applied to a 3d lattice gas model, a generalization of Young's equation for rough and chemically heterogeneous substrates, which combines both Cassie's and Wenzel's laws. We also show that, already in the homogeneous case, the model can exhibit, for particular geometries and appropriated values of the parameters, two regimes governed either by the Wenzel's law or by the Cassie's law. PMID- 11909198 TI - Towards a variational principle for motivated vehicle motion. AB - We deal with the problem of deriving the microscopic equations governing individual car motion based on assumptions about the strategy of driver behavior. We presume the driver behavior to be a result of a certain compromise between the will to move at a speed that is comfortable for him under the surrounding external conditions, comprising the physical state of the road, the weather conditions, etc., and the necessity to keep a safe headway distance between the cars in front of him. Such a strategy implies that a driver can compare the possible ways of further motion and so choose the best one. To describe the driver preferences, we introduce the priority functional whose extremals specify the driver choice. For simplicity we consider a single-lane road. In this case solving the corresponding equations for the extremals we find the relationship between the current acceleration, velocity, and position of the car. As a special case we get a certain generalization of the optimal velocity model similar to the "intelligent driver model" proposed by Treiber and Helbing. PMID- 11909199 TI - Conformal map modeling of the pinning transition in Laplacian growth. AB - In Laplacian growth processes pinning may be expected due to a nonlinear response of a material during dielectric breakdown, or due to stick-slip boundary conditions in two-fluid flow in a porous medium, while thermal noise will lead to depinning. Using a method recently proposed by Hastings and Levitov, the size R(max) approximately E(-alpha)(c) of the pinned pattern is shown to scale with the critical field E(c) (electric field for dielectric breakdown, pressure gradient for fluid flow). These pinned patterns have a lower effective fractal dimension d(f) than diffusion-limited aggregation due to the enhancement of growth at the hot tips of the developing pattern. At finite temperature, thermal noise leads to depinning and growth of patterns with a shape and dimensionality dependent on both E(c) and the thermal noise. Using multifractal analysis, scaling expressions are established for this dependency. PMID- 11909200 TI - Log-periodic route to fractal functions. AB - Log-periodic oscillations have been found to decorate the usual power-law behavior found to describe the approach to a critical point, when the continuous scale-invariance symmetry is partially broken into a discrete-scale invariance symmetry. For Ising or Potts spins with ferromagnetic interactions on hierarchical systems, the relative magnitude of the log-periodic corrections are usually very small, of order 10(-5). In growth processes [diffusion limited aggregation (DLA)], rupture, earthquake, and financial crashes, log-periodic oscillations with amplitudes of the order of 10% have been reported. We suggest a "technical" explanation for this 4 order-of-magnitude difference based on the property of the "regular function" g(x) embodying the effect of the microscopic degrees of freedom summed over in a renormalization group (RG) approach F(x)=g(x)+mu(-1)F(gamma x) of an observable F as a function of a control parameter x. For systems for which the RG equation has not been derived, the previous equation can be understood as a Jackson q integral, which is the natural tool for describing discrete-scale invariance. We classify the "Weierstrass-type" solutions of the RG into two classes characterized by the amplitudes A(n) of the power-law series expansion. These two classes are separated by a novel "critical" point. Growth processes (DLA), rupture, earthquake, and financial crashes thus seem to be characterized by oscillatory or bounded regular microscopic functions that lead to a slow power-law decay of A(n), giving strong log-periodic amplitudes. If in addition, the phases of A(n) are ergodic and mixing, the observable presents self-affine nondifferentiable properties. In contrast, the regular function of statistical physics models with "ferromagnetic"-type interactions at equilibrium involves unbound logarithms of polynomials of the control variable that lead to a fast exponential decay of A(n) giving weak log periodic amplitudes and smoothed observables. PMID- 11909201 TI - Thermodynamics of ionic microgels. AB - We present a theory of dilute aqueous suspensions of microgel particles. It is found that as the number of charged monomers in the polymer network composing mesoscopic gel increases, the particles undergo a swelling transition. Depending on the hydrophobicity of the polymer, this transition can be either continuous or discontinuous. Furthermore, similar to charge stabilized colloidal particles, we find that the electrophoretic mobility of the microgel is controlled by an effective charge. Unlike the colloids, however, for which the effective charge grows asymptotically with the logarithm of the bare charge, the effective charge of an ionic microgel scales as Z(eff) approximately Z0.5. The findings are in good agreement with the experimental measurements. PMID- 11909202 TI - Universality class of discrete solid-on-solid limited mobility nonequilibrium growth models for kinetic surface roughening. AB - We investigate, using the noise reduction technique, the asymptotic universality class of the well-studied nonequilibrium limited mobility atomistic solid-on solid surface growth models introduced by Wolf and Villain (WV) and Das Sarma and Tamborenea (DT) in the context of kinetic surface roughening in ideal molecular beam epitaxy. We find essentially all the earlier conclusions regarding the universality class of DT and WV models to be severely hampered by slow crossover and extremely long-lived transient effects. We identify the correct asymptotic universality class(es) that differs from earlier conclusions in several instances. PMID- 11909203 TI - Quantum refrigeration cycles using spin-1/2 systems as the working substance. AB - The cycle model of a quantum refrigerator composed of two isothermal and two isomagnetic field processes is established. The working substance in the cycle consists of many noninteracting spin-1/2 systems. The performance of the cycle is investigated, based on the quantum master equation and semigroup approach. The general expressions of several important performance parameters, such as the coefficient of performance, cooling rate, and power input, are given. Especially, the case at high temperatures is analyzed in detail. The results obtained are further generalized and discussed, so that they may be directly used to describe the performance of the quantum refrigerator using spin-J systems as the working substance. Finally, the optimum characteristics of the quantum Carnot refrigerator are derived simply. PMID- 11909204 TI - Entropy of electromagnetic polarization. AB - The entropy of electromagnetic polarization is considered in this paper. It is shown that unless the non-field entropy, and not the total entropy, is used as the independent variable in the expression for the internal energy, the first law is violated and the meaning of heat flow, as given by the second law, is contradicted. The total entropy and its field and non-field components are shown to be state functions. The field entropy comprises contributions from the field generated by the contents of the system and stored within as well as outside its boundaries. The contribution of the field stored outside the system boundaries is derived and demonstrated for the case of a uniformly polarized sphere. Finally, expressions are derived for field entropies and entropy densities, in composite systems, using the concept of interaction entropy. The results are shown to be fundamentally different compared to those used in the current literature. PMID- 11909205 TI - Self-organized critical system with no stationary attractor state. AB - A simple model economy with interacting producers and consumers is introduced. When driven by extreme dynamics, the model self-organizes not to an attractor state, but to an asymptote, on which the economy has a constant rate of deflation, is critical, and exhibits avalanches of activity with power-law distributed sizes. This example demonstrates that self-organized critical behavior occurs in a larger class of systems than so far considered: systems not driven to an attractive fixed point, but, e.g., an asymptote, may also display self-organized criticality. PMID- 11909206 TI - Semiclassical spatial correlations in chaotic wave functions. AB - We study the spatial autocorrelation of energy eigenfunctions psi(n)(q) corresponding to classically chaotic systems in the semiclassical regime. Our analysis is based on the Weyl-Wigner formalism for the spectral average C(epsilon)(q(+),q(-),E) of psi(n)(q(+))psi(*)(n)(q(-)), defined as the average over eigenstates within an energy window epsilon centered at E. In this framework C(epsilon) is the Fourier transform in the momentum space of the spectral Wigner function W(x,E;epsilon). Our study reveals the chord structure that C(epsilon) inherits from the spectral Wigner function showing the interplay between the size of the spectral average window, and the spatial separation scale. We discuss under which conditions is it possible to define a local system independent regime for C(epsilon). In doing so, we derive an expression that bridges the existing formulas in the literature and find expressions for C(epsilon)(q(+),q(-),E) valid for any separation size /q(+)-q(-)/. PMID- 11909207 TI - Transition from intermittency to periodicity in lag synchronization in coupled Rossler oscillators. AB - The dynamical and statistical behavior of lag synchronization in two coupled self sustained chaotic Rossler oscillators is reexamined. The lack of uniqueness in the conventional characterization of lag synchronization based on the similarity function has caused much skepticism about the existence of lag synchronization. We provide an evidence that the emergence of lag synchronization is associated with the transition from on-off intermittency to a periodic structure in the laminar phase distribution. PMID- 11909208 TI - Spatiotemporal stability and control of one-way open coupled Lorenz systems. AB - We investigate the spatiotemporal stability of a homogeneous solution in one-way open coupled Lorenz systems, and suppress the spatial instability in the systems by using the H(infinity) control technique. The suppression is illustrated with numerical simulations. In addition, it is shown that the suppression can be also achieved for one-way ring-type systems. PMID- 11909209 TI - Lotka-Volterra system in a random environment. AB - Classical Lotka-Volterra (LV) model for oscillatory behavior of population sizes of two interacting species (predator-prey or parasite-host pairs) is conservative. This may imply unrealistically high sensitivity of the system's behavior to environmental variations. Thus, a generalized LV model is considered with the equation for preys' reproduction containing the following additional terms: quadratic "damping" term that accounts for interspecies competition, and term with white-noise random variations of the preys' reproduction factor that simulates the environmental variations. An exact solution is obtained for the corresponding Fokker-Planck-Kolmogorov equation for stationary probability densities (PDF's) of the population sizes. It shows that both population sizes are independent gamma-distributed stationary random processes. Increasing level of the environmental variations does not lead to extinction of the populations. However it may lead to an intermittent behavior, whereby one or both population sizes experience very rare and violent short pulses or outbreaks while remaining on a very low level most of the time. This intermittency is described analytically by direct use of the solutions for the PDF's as well as by applying theory of excursions of random functions and by predicting PDF of peaks in the predators' population size. PMID- 11909210 TI - Second moment of the Husimi distribution as a measure of complexity of quantum states. AB - We propose the second moment of the Husimi distribution as a measure of complexity of quantum states. The inverse of this quantity represents the effective volume in phase space occupied by the Husimi distribution, and has a good correspondence with chaoticity of classical system. Its properties are similar to the classical entropy proposed by Wehrl, but it is much easier to calculate numerically. We calculate this quantity in the quartic oscillator model, and show that it works well as a measure of chaoticity of quantum states. PMID- 11909211 TI - Peculiarities of the relaxation to an invariant probability measure of nonhyperbolic chaotic attractors in the presence of noise. AB - We study the relaxation to an invariant probability measure on quasihyperbolic and nonhyperbolic chaotic attractors in the presence of noise. We also compare different characteristics of the rate of mixing and show numerically that the rate of mixing for nonhyperbolic chaotic attractors can significantly change under the influence of noise. A mechanism of the noise influence on mixing is presented, which is associated with the dynamics of the instantaneous phase of chaotic trajectories. We also analyze how the synchronization effect can influence the rate of mixing in a system of two coupled chaotic oscillators. PMID- 11909212 TI - Asymmetric unimodal maps at the edge of chaos. AB - We numerically investigate the sensitivity to initial conditions of asymmetric unimodal maps x(t+1)=1-a/x(t)/(z(i)) (i=1,2 correspond to x(t)>0 and x(t)<0, respectively, z(i)>1, 0>1 there are no oscillations at all for the survival probability, and there is almost an exponential decay with the characteristic time determined by Fermi golden rule. In this case, one may not restrict himself to only resonance pair levels. The number of levels perturbed by tunneling grows proportionally to square root of [Lambda] (in other words, instead of isolated pairs there appear the resonance regions containing the sets of strongly coupled levels). In the region of intermediate values of Lambda one has a crossover between both limiting cases, namely, the exponential decay with subsequent long period recurrent behavior. PMID- 11909223 TI - Naturally invariant measure of chaotic attractors and the conditionally invariant measure of embedded chaotic repellers. AB - We study local and global correlations between the naturally invariant measure of a chaotic one-dimensional map f and the conditionally invariant measure of the transiently chaotic map f(H). The two maps differ only within a narrow interval H, while the two measures significantly differ within the images f(l)(H), where l is smaller than some critical number l(c). We point out two different types of correlations. Typically, the critical number l(c) is small. The chi(2) value, which characterizes the global discrepancy between the two measures, typically obeys a power-law dependence on the width epsilon of the interval H, with the exponent identical to the information dimension. If H is centered on an image of the critical point, then l(c) increases indefinitely with the decrease of epsilon, and the chi(2) value obeys a modulated power-law dependence on epsilon. PMID- 11909224 TI - Nonperturbative and perturbative parts of energy eigenfunctions: a three-orbital schematic shell model. AB - We study the division of components of energy eigenfunctions, as the expansion of perturbed states in unperturbed states, into nonperturbative and perturbative parts in a three-orbital schematic shell model possessing a chaotic classical limit, the Hamiltonian of which is composed of a Hamiltonian of noninteracting particles and a perturbation. The perturbative parts of eigenfunctions are expanded in a convergent perturbation expansion by making use of the nonperturbative parts. The division is shown to have the property that, when the underlying classical system is chaotic, the statistics of the components of the nonperturbative parts whose relative localization length are close to 1 is in agreement with the prediction of random-matrix theory. When the underlying classical system is mixed, main bodies of most of the eigenfunctions are found to occupy parts of their nonperturbative regions, with some of the rest of the eigenfunctions being "ergodic" in their nonperturbative regions due to avoided level crossings. In case of the classical system being chaotic, most of the eigenfunctions are found "ergodic" or almost "ergodic" in their nonperturbative regions. Numerical results show that the average relative localization length of nonperturbative parts of eigenfunctions is useful in characterizing the behavior of the quantum system in the process of the underlying classical system changing from a mixed system to a chaotic one. PMID- 11909225 TI - Shadowing breakdown and large errors in dynamical simulations of physical systems. AB - Simulations play a crucial role in the modern study of physical systems. A major open question for long dynamical simulations of physical processes is the role of discretization and truncation errors in the outcome. A general mechanism is described that can cause extremely small noise inputs to result in errors in simulation statistics that are several orders of magnitude larger. A scaling law for the size of such errors in terms of the noise level and properties of the dynamics is given. This result brings into question trajectory averages that are computed for systems with particular dynamical behaviors, in particular, systems that exhibit fluctuating Lyapunov exponents or unstable dimension variability. PMID- 11909226 TI - Pseudointegrable Andreev billiard. AB - A circular Andreev billiard in a uniform magnetic field is studied. It is demonstrated that the classical dynamics is pseudointegrable in the same sense as for rational polygonal billiards. The relation to a specific polygon, the asymmetric barrier billiard, is discussed. Numerical evidence is presented indicating that the Poincare map is typically weak mixing on the invariant sets. This link between these different classes of dynamical systems throws some light on the proximity effect in chaotic Andreev billiards. PMID- 11909227 TI - Experimental observation of the characteristic relations of type-I intermittency in the presence of noise. AB - Recently, it has been reported that the characteristic relation of type-I intermittency in the presence of noise is deformed nontrivially as the channel width epsilon changes from the positive region to the negative. In order to verify it experimentally as a real phenomenon, we study the characteristic relations both for epsilon<0 and for epsilon>0 in a simple inductor-resistor diode circuit that is under noisy circumstances. The experimental results agree well with the theoretical expectation that the characteristic relations are proportional to epsilon(-1/4) for epsilon>0 and proportional to exp(alpha/epsilon/(3/2)) for epsilon<0. PMID- 11909228 TI - Turbulence in small-world networks. AB - The transition to turbulence via spatiotemporal intermittency is investigated in the context of coupled maps defined on small-world networks. The local dynamics is given by the Chate-Manneville minimal map previously used in studies of spatiotemporal intermittency in ordered lattice. The critical boundary separating laminar and turbulent regimes is calculated on the parameter space of the system, given by the coupling strength and the rewiring probability of the network. Windows of relaminarization are present in some regions of the parameter space. New features arise in small-world networks; for instance, the character of the transition to turbulence changes from second-order to a first-order phase transition at some critical value of the rewiring probability. A linear relation characterizing the change in the order of the phase transition is found. The global quantity used as order parameter for the transition also exhibits nontrivial collective behavior for some values of the parameters. These models may describe several processes occurring in nonuniform media where the degree of disorder can be continuously varied through a parameter. PMID- 11909229 TI - Pattern formation in two-frequency forced parametric waves. AB - We present an experimental investigation of superlattice patterns generated on the surface of a fluid via parametric forcing with two commensurate frequencies. The spatiotemporal behavior of four qualitatively different types of superlattice patterns is described in detail. These states are generated via a number of different three-wave resonant interactions. They occur either as symmetry breaking bifurcations of hexagonal patterns composed of a single unstable mode or via nonlinear interactions between the two primary unstable modes generated by the two forcing frequencies. A coherent picture of these states together with the phase space in which they appear is presented. In addition, we describe a number of new superlattice states generated by four-wave interactions that arise when symmetry constraints rule out three-wave resonances. PMID- 11909230 TI - Synchronization: stability and duration time. AB - We consider the problem of stability and duration of the synchronization process between self-excited oscillators, both in their regular and chaotic states. Making use of the properties of Hill equation describing the deviation between the slave and the master, we derive the stability conditions and expressions of the synchronization time. A fairly good agreement is obtained between the analytical and numerical results. PMID- 11909231 TI - Synchronization of chaotic systems with different order. AB - The chaotic synchronization of third-order systems and second-order driven oscillator is studied in this paper. Such a problem is related to synchronization of strictly different chaotic systems. We show that dynamical evolution of second order driven oscillators can be synchronized with the canonical projection of a third-order chaotic system. In this sense, it is said that synchronization is achieved in reduced order. Duffing equation is chosen as slave system whereas Chua oscillator is defined as master system. The synchronization scheme has nonlinear feedback structure. The reduced-order synchronization is attained in a practical sense, i.e., the difference e=x(3)-x(1)(') is close to zero for all time t> or =t(0)> or =0, where t(0) denotes the time of the control activation. PMID- 11909232 TI - Controlling dynamics in spatially extended systems. AB - Spatially extended systems exhibit a variety of spatiotemporal dynamics--from stable to chaotic. These dynamics can change under pathological conditions and impair normal functions. Thus, having the ability to control the altered dynamics for improved functioning has the potential for wide ranging applications in real and artificial systems. Here we propose a simple and general method that can be used to target the spatiotemporal dynamics, both globally and in spatially localized regions, in either direction--i.e., towards the stable or unstable manifold-by simply changing the strength and the sign of an externally applied perturbation or pinning. The method is applicable to both chaotic and nonchaotic systems, with discrete and continuous local dynamics, and for different topologies of interactions. We also apply it to simulate an experiment on epileptogenic neuronal activity in rat hippocampal tissue [B. J. Gluckman et al., J. Neurophys. 76, 6202 (1996)]. This unified approach for differential targeting of global and local dynamics promises to be useful for systems spanning large spatial scales and having structural and functional heterogeneity. PMID- 11909233 TI - Microscopic chaos from Brownian motion in a one-dimensional anharmonic oscillator chain. AB - The problem of relating microscopic chaos to macroscopic behavior in a many degrees-of-freedom system is numerically investigated by analyzing statistical properties associated to the position and momentum of a heavy impurity embedded in a chain of nearest-neighbor anharmonic Fermi-Pasta-Ulam oscillators. For this model we have found that the behavior of the relaxation time of the momentum autocorrelation function of the impurity is different depending on the dynamical regime (either regular or chaotic) of the lattice. PMID- 11909234 TI - Achronal generalized synchronization in mutually coupled semiconductor lasers. AB - Heil et al. [Phys. Rev. Lett. 86, 795 (2001)] recently discovered achronal synchronization of chaos in mutually coupled semiconductor lasers. This paper offers an analytic interpretation of their experiment using a simple rate equation model. Local eigenvalue analysis shows that isochronal synchronization is unstable; achronal synchronization, on the other hand, is stable if a generalized synchronization function is introduced. Single- and multimode simulations have substantiated this rate equation interpretation. Finally, there is a brief examination of "chaos pass filtering." PMID- 11909235 TI - Bifurcations of synchronized responses in synaptically coupled Bonhoffer-van der Pol neurons. AB - The Bonhoffer-van der Pol (BvdP) equation is considered as an important model for studying dynamics in a single neuron. In this paper, we investigate bifurcations of periodic solutions in model equations of four and five BvdP neurons coupled through the characteristics of synaptic transmissions with a time delay. The model can be considered as a dynamical system whose solution includes jumps depending on a condition related to the behavior of the trajectory. Although the solution is discontinuous, we can define the Poincare map as a synthesis of successive submaps, and give its derivatives for obtaining periodic points and their bifurcations. Using our proposed numerical method, we clarify mechanisms of bifurcations among synchronized oscillations with phase-locking patterns by analyzing periodic solutions observed in the coupling system and its subsystems. Moreover, we show that a global behavior of chaotic itinerancy or a phenomenon of chaotic transitions among several quasiattracting states can be observed in higher-dimensional systems of the synaptically four and five coupled neurons. PMID- 11909236 TI - Arithmetical signatures of the dynamics of the Henon map. AB - We report a fourth-degree polynomial that parametrizes analytically all period-4 orbits of the Henon map and use it to investigate arithmetical signatures of the symbolic coding for this prototypical multidimensional system. A discontinuity in the symbolic dynamics observed by Hansen while following numerically a period-6 orbit along a closed loop in parameter space is shown to exist already for period 4. We obtain an analytical expression for the locus of all such discontinuities in parameter space and explain their origin. Our analytical results allow the accurate location of all discontinuities, in contrast with topological methods based on homoclinic tangencies that exist over continuous intervals. PMID- 11909237 TI - Amplitude envelope synchronization in coupled chaotic oscillators. AB - A peculiar type of synchronization has been found when two Van der Pol-Duffing oscillators, evolving in different chaotic attractors, are coupled. As the coupling increases, the frequencies of the two oscillators remain different, while a synchronized modulation of the amplitudes of a signal of each system develops, and a null Lyapunov exponent of the uncoupled systems becomes negative and gradually larger in absolute value. This phenomenon is characterized by an appropriate correlation function between the returns of the signals, and interpreted in terms of the mutual excitation of new frequencies in the oscillators power spectra. This form of synchronization also occurs in other systems, but it shows up mixed with or screened by other forms of synchronization, as illustrated in this paper by means of the examples of the dynamic behavior observed for three other different models of chaotic oscillators. PMID- 11909238 TI - Scaling of structure functions in homogeneous shear-flow turbulence. AB - We apply spectral dynamics and non-Gaussian statistical model of velocity difference to study the scaling of structure functions in homogeneous shear-flow turbulence. Let L(S) be the shear length scale and eta the viscous scale. It is found that, when L(S)/eta is finite, due to a combined effect of viscosity and mean shear, the scaling deviates from normal scaling, and the deviation increases as L(S)/eta decreases. In the presence of a strong shear (L(S)/eta<100), the deviation is substantially larger than the prediction of typical intermittency models, in agreement with recent experiments. As L(S)/eta-->infinity, the normal scaling is valid in the inertial range where viscous and shear effects are negligible. PMID- 11909239 TI - Circulation-strain sum rule in stochastic magnetohydrodynamics. AB - We study probability density functions (PDFs) of the circulation of velocity and magnetic fields in magnetohydrodynamics, computed for a circular contour within inertial range scales. The analysis is based on the instanton method as adapted to the Martin-Siggia-Rose field theory formalism. While in the viscous limit the expected Gaussian behavior of fluctuations is indeed verified, the case of vanishing viscosity is not suitable of a direct saddle-point treatment. To study the latter limit, we take into account fluctuations around quasistatic background fields, which allows us to derive a sum rule relating PDFs of the circulation observables and the rate of the strain tensor. A simple inspection of the sum rule definition leads straightforwardly to the algebraic decay rho(Gamma) 1/Gamma(2) at the circulation PDF tails. PMID- 11909240 TI - Magnetic fluctuations with a zero mean field in a random fluid flow with a finite correlation time and a small magnetic diffusion. AB - Magnetic fluctuations with a zero mean field in a random flow with a finite correlation time and a small yet finite magnetic diffusion are studied. Equation for the second-order correlation function of a magnetic field is derived. This equation comprises spatial derivatives of high orders due to a nonlocal nature of magnetic field transport in a random velocity field with a finite correlation time. For a random Gaussian velocity field with a small correlation time the equation for the second-order correlation function of the magnetic field is a third-order partial differential equation. For this velocity field and a small magnetic diffusion with large magnetic Prandtl numbers the growth rate of the second moment of magnetic field is estimated. The finite correlation time of a turbulent velocity field causes an increase of the growth rate of magnetic fluctuations. It is demonstrated that the results obtained for the cases of a small yet finite magnetic diffusion and a zero magnetic diffusion are different. PMID- 11909241 TI - Continuum description of rarefied gas dynamics. III. The structures of shock waves. AB - We use the one-dimensional steady version of the equations derived in paper I to compute the structure of shock waves and find good agreement with experiment. PMID- 11909242 TI - Critical "dimension" in shell model turbulence. AB - We investigate the Gledzer-Ohkitani-Yamada (GOY) shell model within the scenario of a critical dimension in fully developed turbulence. By changing the conserved quantities, one can continuously vary an "effective dimension" between d=2 and d=3. We identify a critical point between these two situations where the flux of energy changes sign and the helicity flux diverges. Close to the critical point the energy spectrum exhibits a turbulent scaling regime followed by a plateau of thermal equilibrium. The corrections due to intermittency persist close to the critical point. We identify scaling laws and perform a rescaling argument to derive a relation between the critical exponents. We further discuss the distribution function of the energy flux. PMID- 11909243 TI - Flow pattern exchange in the Taylor-Couette system with a very small aspect ratio. AB - Numerical investigation is carried out on the flow pattern exchanges found in Taylor-Couette flows between two concentric rotating cylinders. The inner cylinder rotates while the outer cylinder and both end walls are stationary. The aspect ratio (column length/gap width) is small, and its range is from 0.5 to 1.6. Previous experimental results for this range of the aspect ratio showed that the steady flow patterns are classified into three groups: the normal two-cell mode, anomalous one-cell mode and twin-cell mode. All modes found by experiments are predicted in the present numerical calculation. Besides these three flow modes, an unsteady mode is predicted, which is time dependent and fully developed. The existence of the unsteady mode is also confirmed by our experiments. When the inner cylinder starts to rotate from rest, vortices at the corners of the inner cylinder and both end walls develop, and they induce the normal two-cell mode. The flow of the anomalous one-cell mode or twin-cell mode appears after an abrupt breakdown of symmetric two-cell mode flows. During the gradual deceleration of the inner cylinder, the transitions of flow modes occur. We observed mode transitions between the normal two-cell mode and anomalous one cell mode and mode transitions from the twin-cell mode to the normal two-cell mode, anomalous one-cell mode, and unsteady mode. The critical loci where these mode transitions appear are determined. The numerical confirmation of the twin cell mode is a different result obtained in the present study. PMID- 11909244 TI - Stability analysis of boundary layer flow due to the presence of a small hole on a surface. AB - A linear, temporal, and viscous stability analysis of the boundary layer induced on a solid plane by a three-dimensional potential sink flow is considered. The flow is inviscidly (neutrally) stable. For axisymmetric perturbations, one can analyze separately the stability of those perturbations with a purely circumferential motion, and those with no azimuthal velocity. The first ones are shown to be always stable, a result that is found analytically. The second ones become unstable in a range of (high) Reynolds numbers that depends on the radial wave number. Finally, it is shown that all nonaxisymmetric perturbations are linearly stable. PMID- 11909245 TI - Nonlinear wave dynamics in Faraday instabilities. AB - Nonlinear wave dynamics in parametrically driven surface waves are studied in numerical simulations of the two-dimensional Navier-Stokes equation, with an emphasis on the evolution and interaction between different wave number modes. The dynamics are found to be closely correlated with the single-mode nonlinear saturated wave amplitudes. Modulating behavior of primary wave modes in a particular parameter range and in time scales much longer than the underlining wave periods is observed. PMID- 11909246 TI - Nonhydrodynamic modes and a priori construction of shallow water lattice Boltzmann equations. AB - Lattice Boltzmann equations for the isothermal Navier-Stokes equations have been constructed systematically using a truncated moment expansion of the equilibrium distribution function from continuum kinetic theory. Applied to the shallow water equations, with its different equation of state, the same approach yields discrete equilibria that are subject to a grid scale computational instability. Different and stable equilibria were previously constructed by Salmon [J. Marine Res. 57, 503 (1999)]. The two sets of equilibria differ through a nonhydrodynamic or "ghost" mode that has no direct effect on the hydrodynamic behavior derived in the slowly varying limit. However, Salmon's equilibria eliminate a coupling between hydrodynamic and ghost modes, one that leads to instability with a growth rate increasing with wave number. Previous work has usually assumed that truncated moment expansions lead to stable schemes. Such instabilities have implications for lattice Boltzmann equations that simulate other nonideal equations of state, or that simulate fully compressible, nonisothermal fluids using additional particles. PMID- 11909247 TI - Nonlinear effects due to gravity in a conical Hele-Shaw cell. AB - In this work we study the viscous fingering instability in a conical Hele-Shaw cell under the presence of gravity. We focus on understanding how the dynamical evolution of the fingering patterns is affected by the combined action of gravity and cell topology. Gravity-induced nonlinear effects are studied by a mode coupling approach. Our results show that the interplay between gravity and cell topology leads to important effects, and profoundly modifies pattern evolution. We have found that the most dramatic consequences refer to finger tip behavior. Depending on the relative values of fluids' densities and viscosities, finger tip splitting reaches maximum intensity at well defined, preferred values of the cell opening angle. In fact, finger tip splitting can be completely replaced by finger tip sharpening as the cell angle is varied. Finger competition dynamics is also significantly changed: it is considerably enhanced (restrained) if the displaced fluid is more (less) dense. PMID- 11909248 TI - Nonlinear states of the screw dynamo. AB - The self-excitation of magnetic field by a spiral Couette flow between two coaxial cylinders is considered. We solve numerically the fully nonlinear, three dimensional magnetohydrodynamic (MHD) equations for magnetic Prandtl numbers P(m) (ratio of kinematic viscosity to magnetic diffusivity) between 0.14 and 10 and kinematic and magnetic Reynolds numbers up to about 2000. In the initial stage of exponential field growth (kinematic dynamo regime), we find that the dynamo switches from one distinct regime to another as the radial width delta(r)(B) of the magnetic field distribution becomes smaller than the separation of the field maximum from the flow boundary. The saturation of magnetic field growth is due to a reduction in the velocity shear resulting mainly from the azimuthally and axially averaged part of the Lorentz force, which agrees with an asymptotic result for the limit of P(m)<<1. In the parameter regime considered, the magnetic energy decreases with kinematic Reynolds number as Re-0.84, which is approximately as predicted by the nonlinear asymptotic theory (approximately Re( 1)). However, when the velocity field is maintained by a volume force (rather than by viscous stress) the dependence of magnetic energy on the kinematic Reynolds number is much weaker. PMID- 11909250 TI - Memory effects in a turbulent dynamo: generation and propagation of a large-scale magnetic field. AB - We are concerned with large-scale magnetic-field dynamo generation and propagation of magnetic fronts in turbulent electrically conducting fluids. An effective equation for the large-scale magnetic field is developed here that takes into account the finite correlation times of the turbulent flow. This equation involves the memory integrals corresponding to the dynamo source term describing the alpha effect and turbulent transport of magnetic field. We find that the memory effects can drastically change the dynamo growth rate, in particular, nonlocal turbulent transport might increase the growth rate several times compared to the conventional gradient transport expression. Moreover, the integral turbulent transport term leads to a large decrease of the speed of magnetic front propagation. PMID- 11909249 TI - Flow paths in wetting unsaturated flow: experiments and simulations. AB - The flow paths and instabilities of gravity driven infiltration of a wetting fluid into a porous medium are studied. The model experiments and simulations independently represent techniques to study the unsaturated flow in porous media, and they produce a consistent picture of how the paths of fluid transport form and depend on the relative strength of the gravitational force. The experiments, which employ a transparent and quasi-two-dimensional model, reveal that the fluid pathways contain an internal link-blob structure and increase in width with decreasing gravity. The model, which couples the well established invasion percolation model for capillary governed flow with a model that describes the viscous film flow in partially filled pores, corroborates these experimental findings. PMID- 11909251 TI - Energy and structure of inertial range turbulence deduced from an evolution of fluid impulse. AB - We explore numerically a very simple idea that may provide a material explanation for inertial range turbulence. We base a Lagrangian model of viscous incompressible fluid flow on an evolving ensemble of vortex doublet sheets. Initially these are randomly oriented and randomly distributed within a disk in two-dimensional space. These sheets are then actively transported (in two dimensions) according to the Oseledets equation of motion for fluid impulse. The mutual interaction of these sheets, and their diffusion, establishes a velocity fluctuation field. In a specific sense this evolution is self-affine, and we exploit this property to calculate standard statistical measures for the fluctuation field. We determine from this simple model the second-order structure function and the energy spectrum of inertial range turbulence. PMID- 11909252 TI - Growth of binary fluid convection: role of the concentration field. AB - The growth of convection in binary fluid mixtures out of different perturbations of the quiescent conductive state is investigated using finite-difference numerical simulations for realistic ethanol-water parameters with strong negative Soret coupling between temperature and concentration fluctuations. Several different analysis tools are used to elucidate the complex spatiotemporal behavior associated with the dramatic concentration redistribution during the transients. It shows first the competition between counterpropagating waves that initially superimpose to form standing wave perturbations. Having reached a critical amplitude an advective breaking of the concentration wave triggers a very fast flow-induced transition from standing to traveling wave convection with large phase velocity and large concentration field amplitudes. Strongly nonlinear advective mixing and weak long-time diffusive homogenization then slow down the waves. PMID- 11909253 TI - Experiments on free decay of quasi-two-dimensional turbulent flows. AB - Decaying quasi-two-dimensional turbulence in a thin-layer flow is explored in laboratory experiments. We report the presence of power-law interval in the enstrophy decay law, in agreement with earlier experiments by Cardoso et al. [Phys. Rev. E 49, 454 (1994)] and Hansen et al. [Phys. Rev. E 58, 7261 (1998)]. The decay exponent proves sensitive to the way in which the energy decay is compensated. For the range of initial microscale Reynolds numbers between 35 and 95, the decay exponent is close to -0.4 for the ratio of enstrophy to energy, and to -0.75 for the enstrophy multiplied with a compensating factor of exp( 2lambda(t)), where lambda is the bottom-drag coefficient and t the decay time. The vorticity behavior does not comply with the theory of Carnevale et al. [Phys. Rev. Lett. 66, 2735 (1991)]: robust vortices are not observed in the vorticity field and the vorticity kurtosis is less than the Gaussian value. PMID- 11909254 TI - Vortex filament motion under the localized induction approximation in terms of Weierstrass elliptic functions. AB - We study quasiperiodic solutions of the localized induction approximation in terms of the elliptic functions of Weierstrass. They describe the Kida-class motion of a thin vortex filament in an incompressible inviscid fluid. Our solution includes various filament shapes such as the vortex ring, the helicoidal filament, the plane sinusoidal filament, and the Hasimoto type-1 soliton filament. PMID- 11909255 TI - Lattice Boltzmann simulation of chemical dissolution in porous media. AB - In this paper, we develop a lattice Boltzmann model for simulating the transport and reaction of fluids in porous media. To simulate such a system, we account for the interaction of forced convection, molecular diffusion, and surface reaction. The problem is complicated by the evolution of the porous media geometry due to chemical reactions, which may significantly and continuously modify the hydrologic properties of the media. The particular application that motivates the present study is acid stimulation, a common technique used to increase production from petroleum reservoirs. This technique involves the injection of acid (e.g., hydrochloric acid, HCl, acetic acid, HAc) into the formation to dissolve minerals comprising the rock. As acid is injected, highly conductive channels or "wormholes" may be formed. The dissolution of carbonate rocks in 0.5M HCl and 0.5M HAc is simulated with the lattice Boltzmann model developed in this study. The dependence of dissolution process and the geometry of the final wormhole pattern on the acid type and the injection rate is studied. The results agree qualitatively with the experimental and theoretical analyses of others and substantiate the previous finding that there exists an optimal injection rate at which the wormhole is formed as well as the number of pore volumes of the injected fluid to break through is minimized. This study also confirms the experimentally observed phenomenon that the optimal injection rate decreases and the corresponding minimized number of pore volumes to break through increases as the acid is changed from HCl to HAc. Simulations suggest that the proposed lattice Boltzmann model may serve as an alternative reliable quantitative approach to study chemical dissolution in porous media. PMID- 11909256 TI - Steady ion momentum in nonlinear plasma waves. AB - The analysis of a one-dimensional two-fluid hydrodynamic model with relativistic electrons and nonrelativistic ions shows that the propagation of a nonlinear plasma wave is accompanied by a steady currentless plasma drift. Ions, due to their larger mass, appear to be the main carriers of the average momentum of the plasma wave. Two examples of nonlinear plasma waves generated by moving sources (short laser pulses and electron bunches) are analyzed to show details of the energy and momentum conservation laws. PMID- 11909257 TI - Theory of K(alpha) generation by femtosecond laser-produced hot electrons in thin foils. AB - An analytical model of femtosecond K(alpha) x-ray generation from laser irradiated foils is presented. Expressions are found for the photon emission yield in both forward and backward directions in integral form as a function of hot-electron temperature and target thickness. It is found that for any given target material, there is a foil thickness and a hot-electron temperature at which the K(alpha) emission is maximized. Conversion efficiencies are consistent with contemporary measurements of K(alpha) radiation produced with femtosecond lasers. PMID- 11909258 TI - Spatiotemporal characteristics of the collisionless rf sheath and the ion energy distributions arriving at rf-biased electrodes. AB - A self-consistent fluid model is proposed for describing collisionless radio frequency (rf) sheaths driven by a sinusoidal current source. This model includes all time-dependent terms in ion fluid equations, which are commonly ignored in some analytical models and numerical simulations. Moreover, an equivalent circuit model is introduced to determine self-consistently the relationship between the instantaneous potential at a rf-biased electrode and the sheath thickness. The time-dependent voltage wave form, the sheath thickness and the ion flux at the electrode as well as the spatiotemporal variations of the potential, the electric field and the ion density inside the sheath are calculated for various rf powers and ratios of the rf frequency to the ion plasma frequency. The ion energy distributions ( IEDs) impinging on the rf-biased electrode are also calculated with the ion flux at the electrode. The numerical results show that the frequency ratio is a crucial parameter determining the spatiotemporal variations of the rf sheath and the shape of the IEDs. PMID- 11909259 TI - Two-dimensional collision of probe photons with relativistic ionization fronts. AB - The collision of a probe laser pulse with a relativistic ionization front is analyzed via two-dimensional ray-tracing theory and simulations. It is shown that collisions in higher dimensions lead to new regimes for the frequency upshift of the probe photons; the frequency upshift can be considerably higher for particular collision angles that maximize the interaction length with the ionization front gradient. Finite ionization fronts also lead to angle-dependent frequency upshifts, thus acting as diffraction gratings. PMID- 11909260 TI - Energetics of electromagnetic wave transformation in a time-varying magnetoplasma medium. AB - The transformation of a circularly polarized electromagnetic wave in a magnetoplasma medium with increasing plasma density is considered. The wave propagates along the static magnetic field. Complete analysis, including ion motion, is given both for slow (compared to the wave frequency) and rapid ionization rate. In the case of slow temporal variation of the plasma density, a relation between the energy of the wave and its frequency, which is conserved during the plasma creation process (adiabatic invariant), is found. The existence of significant energy losses follows from the invariant. The dissipative mechanism is explained via consideration of the case of a sudden growth of plasma density in time from one value to another. It is shown that energy transforms into the kinetic energy of carriers, and preionization of the medium plays a principal role in the dissipation process. In the special case of a whistler wave, up to 50% of the energy may be transformed into an ion-cyclotron wave when dense plasma is rapidly created. PMID- 11909261 TI - Stopping power of nonideal, partially ionized plasmas. AB - The stopping power of strongly coupled, partially ionized plasmas is investigated for charged beam particles with arbitrary velocities. Our approach is based on kinetic equations of the Boltzmann type that are suitably generalized to describe three-particle collisions. In this way, we consider elastic collisions between the beam and free plasma particles as well as the ionization and excitation of composite plasma particles by beam particle impact. Explicit expressions for both contributions are given in terms of the momentum transfer cross section that has been generalized for three-particle collisions. For fast beam particles, we obtain a generalized Bethe formula that includes correction terms due to the nonideality of the target plasma. Results are shown for hydrogen, carbon, and argon plasmas. Considerable modifications compared to the ideal behavior arise for strongly coupled plasmas. In particular, we are able to describe the Mott transition in the stopping power of dense, partially ionized plasmas. PMID- 11909262 TI - Unified formulation for inhomogeneity-driven instabilities in the lower-hybrid range. AB - A local dispersion relation that describes inhomogeneity-driven instabilities in the lower-hybrid range is derived following a procedure that correctly describes energy exchange between waves and particles in inhomogeneous media, correcting some inherent ambiguities associated with the standard formalism found in the literature. Numerical solutions of this improved dispersion relation show that it constitutes a unified formulation for the instabilities in the lower-hybrid range, describing the so-called modified two-stream instability, excited by the ion cross-field drift, including the ion Weibel instability, and also describing the lower-hybrid drift instability, which is due to inhomogeneity effects on the electron population. PMID- 11909263 TI - Quasilinear diffusion as a result of modulational instability in the pulsar plasma. AB - Quasilinear diffusion due to modulational instability is considered in this paper. Interaction between the high-frequency, nearly transverse O mode (or the transverse X mode) and the low-frequency, nearly longitudinal L-O mode in a pulsar magnetospheric pair plasma can lead to modulational instability. The low frequency L-O mode is superluminal, which is not subjected to usual Landau damping, and it is possible that excess wave energy is stored in this superluminal mode. The superluminal low-frequency L-O mode can dissipate in a way similar to the process of Langmuir wave collapse, that is, it cascades from the long- to short-wavelength regimes. When the phase speed becomes less than c, the waves can be damped through various resonances. We consider, in particular, damping through cyclotron resonance, which can lead to particle acceleration. The energetic beam particles, which have a very small spread initially, can develop a high-energy distribution tail, acquiring pitch angles through quasilinear diffusion. These particles can emit gamma rays through synchrotron radiation, contributing to the observed pulsar high-energy emission. PMID- 11909264 TI - Injection of electron beam into a toroidal trap using chaotic orbits near magnetic null. AB - Injection of charged particle beam into a toroidal magnetic trap enables a variety of interesting experiments on non-neutral plasmas. Stationary radial electric field has been produced in a toroidal geometry by injecting electrons continuously. When an electron gun is placed near an X point of magnetic separatrix, the electron beam spreads efficiently through chaotic orbits, and electrons distribute densely in the torus. The current returning back to the gun can be minimized less than 1% of the total emission. PMID- 11909266 TI - Theory of wave propagation along a waveguide filled with moving magnetized plasma. AB - Making use of the transformation of constitutive relations for electromagnetic waves and the transformation of the wave vector in Minkowski space, we have worked out the theory of wave propagation along a waveguide filled with moving magnetized plasma (MMPW). The dispersion equations of the wave propagation in a circular MMPW are given in this paper, along with a detailed discussion of their behaviors. Numerical calculations show that there are many interesting and important features of the wave propagation along an MMPW compared with that in a stationary magnetized plasma-filled waveguide (MPW). PMID- 11909265 TI - Spatially resolved x-ray spectroscopy investigation of femtosecond laser irradiated Ar clusters. AB - High temperature plasmas have been created by irradiating Ar clusters with high intensity 60-fs laser pulses. Detailed spectroscopic analysis of spatially resolved, high resolution x-ray data near the He(alpha) line of Ar is consistent with a two-temperature collisional-radiative model incorporating the effects of highly energetic electrons. The results of the spectral analysis are compared with a theoretical hydrodynamic model of cluster production, as well as interferometric data. The plasma parameters are notably uniform over one Rayleigh length (600 microm). PMID- 11909267 TI - Electric microfield distributions in electron-ion plasmas. AB - The low-frequency electric microfield distribution in a Coulomb plasma is calculated for various plasma parameters, from weak to strong Coulomb coupling and from zero to strong electron screening. Two methods of numerical calculations are employed: the adjustable-parameter exponential approximation and the Monte Carlo simulation. The results are represented by analytic fitting formulas suitable for applications. PMID- 11909269 TI - Harmonic generation of ultraintense laser pulses in underdense plasma. AB - We propose a nonlinear theory of the generation of harmonic radiation from the interaction of ultraintense laser beams with plasma. The harmonic generation is related to the transition of the laser-plasma equilibrium state. By taking into account correlations among sidebands, we study many sidebands comprehensively. The harmonic generation is viewed as a redistribution of laser field over different frequencies because of the requirement of system stability. We introduce a system parameter S that is related to the sideband intensity spectrum and self-consistently calculate the value of S. Our numerical experiments reveal that the variations of controllable system parameters, plasma density, and laser peak intensity have a great effect on S. PMID- 11909268 TI - Observation of neutron spectrum produced by fast deuterons via ultraintense laser plasma interactions. AB - We report the first precise spectral measurement of fast neutrons produced in a deuterated plastic target irradiated by an ultraintense sub-picosecond laser pulse. The 500-fs, 50-J, 1054-nm laser pulse was focused on the deuterated polystyrene target with an intensity of 2 x 10(19) W/cm(2). The neutron spectra were observed at 55 degrees and 90 degrees to the rear target normal. The neutron emission was 7 x 10(4) per steradian for each detector. The observed neutron spectra prove the acceleration of deuterons and neutron production by d(d,n)3He reactions in the target. The neutron spectra were compared with Monte Carlo simulation results and the deuteron's directional anisotropy and energy spectrum were studied. We conclude that 2% of the laser energy was converted to deuterons, which has an energy range of 30 keV up to 3 MeV. PMID- 11909270 TI - Oblique electrostatic modes in self-gravitating dusty plasmas. AB - The propagation of oblique and perpendicular electrostatic modes in dusty self gravitational magnetized plasmas is treated with due care for the small gravitational effects between charged dust particles and for the correct balance between electrostatic and gravitational forces. At purely oblique propagation, generalizations of the dust-cyclotron and dust-acoustic modes are found, where the latter can be subject to a Jeans instability. For strictly perpendicular propagation, only a mixed dust-acoustic and dust lower-hybrid mode occur at low frequencies. A Jeans collapse can be avoided by strong enough magnetic fields, the criterion for which is given. PMID- 11909271 TI - Axisymmetric relativistic self-channeling of laser light in plasmas. AB - By using an improved cavitation model, relativistic self-channeling structures are derived, which make it possible to propagate laser powers exceeding the critical one for self-focusing. A propagation mode for high laser power is also presented which is qualitatively different from those in the weakly relativistic case. Structural stability analysis shows that stable self-wave-guide propagation can take place. PMID- 11909272 TI - Shear Alfven waves in turbulent plasmas. AB - The rate of decay of shear Alfven waves along a magnetic field line of a diffusive plasma grows with the number of nodes of the initial perturbation. It is reasonable to think that the energy dissipation produced by this decay will be small if the perturbation was localized in a small set. This does not happen in turbulent plasmas: transport of the oscillation by the flow involves the whole domain. A general relation is obtained proving that the global energy dissipation is bounded below by an exponential of the number of nodes of any shear Alfven wave along a segment of any field line of the average magnetic field. PMID- 11909273 TI - Dense plasma temperature equilibration in the binary collision approximation. AB - Temperature equilibration in dense, strongly coupled plasmas has been investigated without most of the usual simplifying assumptions. A quantum kinetic approach is used that accounts for strong electron-ion collisions through an exact T-matrix treatment of the scattering cross section using a screened interaction. Our results reveal the accuracy of the usual Spitzer formula for Coulomb logarithms larger than about three. Moreover, a simple model based on hyperbolic orbits yields surprisingly accurate results. We also have included equation of state effects to describe realistic plasmas. PMID- 11909274 TI - Parametric and resonant transition radiation in periodic stratified structures. AB - We consider the problem of electromagnetic emission when an electrically charged particle crosses a periodically stratified structure following an arbitrary linear trajectory. A theory generalizing a recently developed method [Phys. Rev. E 63, 016613 (2001)] is presented in the framework of the classical theory of electromagnetism in continuous media. It allows one to account for both the so called parametric radiation and the resonant transition radiation. We implement our model to interpret the experiments performed by Kaplin et al. [Appl. Phys. Lett. 76, 3647 (2000)] in the x-ray domain with a stack of 300 W/B(4)C bilayers irradiated by 500 MeV electrons. PMID- 11909275 TI - Approximate solutions of two-way diffusion equations. AB - In this paper, a general and systematic scheme is formulated for finding approximate solutions of two-way diffusion equations. This expansion scheme is valid for arbitrary mean-free path and can be carried out to any desired accuracy. Its potential is demonstrated by constructing approximate solutions for two problems concerning the kinetics of an electron beam, and the accuracy is found to be very good even when only a few terms are included in the expansion. The approximate solutions found are compared with numerical calculations and previous analytical work in the literature. PMID- 11909276 TI - Phase and gain measurements in a distributed-loss cyclotron-resonance maser amplifier. AB - The control of gain and phase delay in a cyclotron-resonance maser (CRM) amplifier is essential for a variety of applications. In this experiment, the gain and phase-delay variations are measured with respect to controlling parameters; the electron-beam current and the axial magnetic field. Following Chu et al. [Phys. Rev. Lett. 74, 1103 (1995)], the CRM amplifier comprises of a distributed-loss waveguide to enable high gain without oscillations. Our experiment yields an amplification up to 26 dB, and a phase-delay control range of 360 degrees. In order to keep a fixed gain with the varying phase delay, the two controlling parameters (i.e., the solenoid field and the beam current) are operated together in a compensating mode. The experiment is conducted in a frequency of 7.3 GHz, with an electron beam of 18-kV voltage and 0.25-0.4-A current. The experimental results are compared with a theoretical model. Practical implementations of gain and phase control in CRM devices are discussed. PMID- 11909277 TI - Analytical description of stripping foil extraction from isochronous cyclotrons. AB - Analytical expressions are derived describing beam parameters on a stripping foil (SF) as a function of radial amplitude of betatron oscillations and of energy gain. The results computed from these expressions are in good agreement with those from numerical calculations. These results indicate the existence of a parametric relationship between radial emittance and energy spread, via the amplitude of radial betatron oscillations. This relationship enables one to generate the working diagram of expected beam parameters on a SF. Such a diagram can be particularly useful for designers of extraction systems, since it gives the relationship between quantities used as extraction system input parameters. The derived analytical expressions can make the design of cyclotrons easier and significantly reduce the need of numerical simulations. PMID- 11909278 TI - Experimental observation of short-pulse upshifted frequency microwaves from a laser-created overdense plasma. AB - A short and frequency upshifted from a source microwave pulse is experimentally generated by the overdense plasma that is rapidly created by a laser. The source wave, whose frequency is 9 GHz, is propagating in the waveguide filled with tetrakis-dimethyl-amino-ethylene gas, which is to be converted to the overdense plasma by the laser. The detected frequency of the pulse is over 31.4 GHz and its duration is 10 ns. This technique has the potential for the generation of a tunable frequency source. PMID- 11909279 TI - Theory of acoustic scattering by supported ridges at a solid-liquid interface. AB - We combine a general Green's function formalism and an approach due to Nyborg [W. L. Nyborg, in Acoustic Streaming, Physical Acoustics, edited by W. P. Mason (Academic, London, 1965), Vol. II B, Chap. 11] to calculate the first-order pressure and second-order pressure gradient fields in the vicinity of solid inhomogeneities at a solid/liquid interface. We treat the problem of scattering of an incident acoustic plane wave by a single ridge and two parallel ridges separated by a trench on a planar substrate. The calculated vibrational density of states shows the existence of resonances at low frequencies, especially in the case of a trench. Excitation of a trench resonant vibrational mode enhances the magnitude of the first-order pressure and of the second-order pressure gradient. The resonant frequencies of a trench decrease and the pressure enhancement increases with increasing aspect ratio of the ridges (height to width). PMID- 11909280 TI - Interferometric methods in diagnostics of polarization singularities. AB - An interferometric technique for analysis of a polarization singular skeleton (s contours and C points) of an optical vector field is elaborated. It was shown that complete characteristics of C points and s contours may be reconstructed from interferometric data. Some examples of elaborated interferometric technique application to the analysis of randomly polarized speckle-fields singularities are presented. PMID- 11909281 TI - Electromigration-induced propagation of nonlinear surface waves. AB - Due to the effects of surface electromigration, waves can propagate over the free surface of a current-carrying metallic or semiconducting film of thickness h(0). In this paper, waves of finite amplitude, and slow modulations of these waves, are studied. Periodic wave trains of finite amplitude are found, as well as their dispersion relation. If the film material is isotropic, a wave train with wavelength lambda is unstable if lambda/h(0)<3.9027 ..., and is otherwise marginally stable. The equation of motion for slow modulations of a finite amplitude, periodic wave train is shown to be the nonlinear Schrodinger equation. As a result, envelope solitons can travel over the film's surface. PMID- 11909282 TI - Time delay distribution in Bragg gratings. AB - A layer-by-layer analysis of the time delay of both reflected and transmitted light in one-dimensional photonic band-gap structures is developed and applied to uniform Bragg gratings. An effective Fabry-Perot cavity is associated with every layer along the Bragg grating, multiple paths with a well defined layer traversal time are identified, and the average time is computed, introducing an appropriate weighting factor that accounts for interference between different paths. The analysis presented leads directly to a complex-valued time delay whose real part is shown to be equivalent to the classic phase time delay. Physical meaning is also given to the imaginary part. The local dwell time, interpreted as the average time spent by light in the layer independently of the final (transmitted or reflected) state, is proved analytically to be related to the energy density distribution when small index change gratings are considered. The time delay evolution is derived at different wavelengths and the nonuniform distribution along the grating is discussed. Nonintuitive features such as superluminal transmission time delay for propagation inside the band gap and negative reflection time delay close to transmission resonances are addressed. Finally, the effect of introducing a small perturbation in the structure is shown to be directly related to the local time delay and is proposed as a possible experimental measurement scheme for both its real and imaginary parts. PMID- 11909283 TI - Multichannel pulse dynamics in a stabilized Ginzburg-Landau system. AB - We study the stability and interactions of chirped solitary pulses in a system of nonlinearly coupled cubic Ginzburg-Landau (CGL) equations with a group-velocity mismatch between them, where each CGL equation is stabilized by linearly coupling it to an additional linear dissipative equation. In the context of nonlinear fiber optics, the model describes transmission and collisions of pulses at different wavelengths in a dual-core fiber, in which the active core is furnished with bandwidth-limited gain, while the other, passive (lossy) one is necessary for stabilization of the solitary pulses. Complete and incomplete collisions of pulses in two channels in the cases of anomalous and normal dispersion in the active core are analyzed by means of perturbation theory and direct numerical simulations. It is demonstrated that the model may readily support fully stable pulses whose collisions are quasielastic, provided that the group-velocity difference between the two channels exceeds a critical value. In the case of quasielastic collisions, the temporal shift of pulses, predicted by the analytical approach, is in semiquantitative agreement with direct numerical results in the case of anomalous dispersion (in the opposite case, the perturbation theory does not apply). We also consider a simultaneous collision between pulses in three channels, concluding that this collision remains quasielastic, and the pulses remain completely stable. Thus, the model may be a starting point for the design of a stabilized wavelength-division-multiplexed transmission system. PMID- 11909284 TI - Suppression of Manakov soliton interference in optical fibers. AB - In this paper, we study the interaction of two vector solitons in the Manakov equations that govern pulse transmission in randomly birefringent fibers. Under the assumptions that these solitons initially are well separated and having nearly the same amplitudes and velocities but arbitrary polarizations, we derive a reduced set of ordinary differential equations for both solitons' parameters. We then solve this reduced system analytically. Our analytical solutions show that, when two Manakov solitons have the same amplitude and phases, their collision distance steadily increases as their initial polarizations change from parallel to orthogonal. In particular, the collision distance at orthogonal polarizations is of the order of the square of the collision distance at parallel polarizations. When the Manakov solitons have different amplitudes, a quasiequidistant bound state can be formed. The degrees of position and amplitude oscillations in this bound state diminish as the initial polarizations change from parallel to orthogonal. With a combination of launching Manakov solitons along orthogonal polarizations and at unequal amplitudes, Manakov-soliton interference is almost completely suppressed. These theoretical results are in excellent agreement with our direct numerical simulations. PMID- 11909285 TI - Purely imaginary eigenvalues of Zakharov-Shabat systems. AB - Zakharov-Shabat systems with single-hump and real, but not necessarily symmetric, potentials are shown to have purely imaginary eigenvalues only. Coupled with examples of double-hump potentials with nonimaginary eigenvalues, this establishes that confinement of Zakharov-Shabat eigenvalues to the imaginary axis is a characteristic of potentials whose energy is concentrated in a single region of the time axis. PMID- 11909286 TI - Anomalous dispersion and superluminal group velocity in a coaxial photonic crystal: theory and experiment. AB - We demonstrate that coaxial cables with a periodic impedance exhibit dispersion properties specific to photonic crystals, albeit on a much lower frequency scale. Highly superluminal (>2c) pulse propagation is observed near the photonic band gap at 10 MHz. The influence of group velocity dispersion and crystal length on the traveling speed and shape of a Gaussian pulse are discussed. Results compare favorably with a simple multilayer theory and a coupled-mass model of the structure. PMID- 11909287 TI - Spatial optical solitons in nonlinear photonic crystals. AB - We study spatial optical solitons in a one-dimensional nonlinear photonic crystal created by an array of thin-film nonlinear waveguides, the so-called Dirac-comb nonlinear lattice. We analyze modulational instability of the extended Bloch-wave modes and also investigate the existence and stability of bright, dark, and "twisted" spatially localized modes in such periodic structures. Additionally, we discuss both similarities and differences of our general results with the simplified models of nonlinear periodic media described by the discrete nonlinear Schrodinger equation, derived in the tight-binding approximation, and the coupled mode theory, valid for shallow periodic modulations of the optical refractive index. PMID- 11909288 TI - Polarization patterns and vectorial defects in type-II optical parametric oscillators. AB - Previous studies of lasers and nonlinear resonators have revealed that the polarization degree of freedom allows for the formation of polarization patterns and novel localized structures, such as vectorial defects. Type- II optical parametric oscillators are characterized by the fact that the down-converted beams are emitted in orthogonal polarizations. In this paper we show the results of the study of pattern and defect formation and dynamics in a type-II degenerate optical parametric oscillator, for which the pump field is not resonated in the cavity. We find that traveling waves are the predominant solutions and that the defects are vectorial dislocations that appear at the boundaries of the regions where traveling waves of different phase or wave-vector orientation are formed. A dislocation is defined by two topological charges, one associated with the phase and another with the wave-vector orientation. We also show how to stabilize a single defect in a realistic experimental situation. The effects of phase mismatch of nonlinear interaction are finally considered. PMID- 11909289 TI - Collective behavior of parametric oscillators. AB - We revisit the mean-field model of globally and harmonically coupled parametric oscillators subject to periodic block pulses with initially random phases. The phase diagram of regions of collective parametric instability is presented, as is a detailed characterization of the motions underlying these instabilities. This presentation includes regimes not identified in earlier work [I. Bena and C. Van den Broeck, Europhys. Lett. 48, 498 (1999)]. In addition to the familiar parametric instability of individual oscillators, two kinds of collective instabilities are identified. In one the mean amplitude diverges monotonically while in the other the divergence is oscillatory. The frequencies of collective oscillatory instabilities in general bear no simple relation to the eigenfrequencies of the individual oscillators nor to the frequency of the external modulation. Numerical simulations show that systems with only nearest neighbor coupling have collective instabilities similar to those of the mean field model. Many of the mean-field results are already apparent in a simple dimer [M. Copelli and K. Lindenberg, Phys. Rev. E 63, 036605 (2001)]. PMID- 11909290 TI - Optical absorption for parallel cylinder arrays. AB - We study the long-wavelength electromagnetic resonances of interacting cylinder arrays. By using a normal-modes expansion where the effects of geometry and material are separated, it is shown that two parallel cylinders with different radii have electromagnetic modes distributed symmetrically about depolarization factor 1 / 2. Both sets couple to longitudinal and transverse components of the external field, but amplitudes of symmetric depolarization factors become exchanged when considering longitudinal or transverse polarization. We also find that amplitudes satisfy sum rules that depend on the ratio of the cylinders radii. The main effect of the difference in radii is a spectral shift towards the isolated cylinder resonance as this difference increases. PMID- 11909291 TI - Trapping of vibrational energy in crumpled sheets. AB - We investigate the propagation of transverse elastic waves in crumpled media. We set up the wave equation for transverse waves on a generic curved, strained surface via a Langrangian formalism and use this to study the scaling behavior of the dispersion curves near the ridges and on the flat facets. This analysis suggests that ridges act as barriers to wave propagation and that modes in a certain frequency regime could be trapped in the facets. A simulation study of the wave propagation qualitatively supported our analysis and showed interesting effects of the ridges on wave propagation. PMID- 11909292 TI - Increased damping of irregular resonators. AB - It is shown that fractal drums and jagged geometry resonators may be more damped than ordinary Euclidean systems. Several damping mechanisms are examined and studied by numerical calculations. The results depend on the dissipation mechanisms but globally they increase with localization, frequency, and the irregularity of the resonator. The increased dissipation is due to the uneven spatial distribution of the vibrational amplitude in two different ways. First, it is related to the partial confinement of the vibrational modes. Secondly, increased dissipation may be due to singularities in the amplitude distribution. This is the case when a few points exist where the vibration is pinned to zero inducing local logarithmic singularities. This last effect can be spectacular: a single defect can dominate the surface damping by viscous forces of a square drum. PMID- 11909293 TI - Body versus surface forces in continuum mechanics: is the Maxwell stress tensor a physically objective Cauchy stress? AB - The Maxwell stress tensor (MST) T(M) plays an important role in the dynamics of continua interacting with external fields, as in the commercially and scientifically important case of "ferrofluids." As a conceptual entity in quasistatic systems, the MST derives from the definition f(M)def=inverted Delta x T(M), where f(M)(x) is a physically objective volumetric external body-force density field at a point x of a continuum, derived from the solution of the pertinent governing equations. Beginning with the fact that T(M) is not uniquely defined via the preceding relationship from knowledge of f(M), we point out in this paper that the interpretation of T(M) as being a physical stress is not only conceptually incorrect, but that in commonly occuring situations this interpretation will result in incorrect predictions of the physical response of the system. In short, by elementary examples, this paper emphasizes the need to maintain the classical physical distinction between the notions of body forces f and stresses T. These examples include calculations of the torque on bodies, the work required to deform a fluid continuum, and the rate of interchange of energy between mechanical and other modes. PMID- 11909294 TI - Spatially resolved photonic transfer through mesoscopic heterowires. AB - We report spatially resolved observations of light wave propagation along high refraction index dielectric heterowires lying on a transparent substrate. The heterowires are made of linear chains of closely packed mesoscopic particles. The optical excitation of these heterowires is performed through channel waveguides featuring submicrometer transverse cross sections. Both numerical simulations and near-field optical images, recorded with a photon scanning tunneling microscope, agree to show that, at visible frequencies, tuning the periodicity of the heterowires controls the propagation length within a range of several micrometers. PMID- 11909295 TI - Coherent interaction effects in pulses propagating through a doped nonlinear dispersive medium. AB - Using a numerical approach we report on the cloning dynamics of simultaneous self induced transparency (SIT) and nonlinear Schrodinger (NLS) solitons in a doped nonlinear dispersive medium. This technique involves a three-level atomic system interacting resonantly with two optical fields within a Lambda scheme. As a result, a pulse in the signal frequency is transformed into a replica of the pulse in the pump frequency. The atomic population evolution shows that the basic mechanism behind the cloning process is the coherent population trapping effect. Furthermore, it is shown that the signal clone presents characteristics of both the SIT soliton and NLS soliton. PMID- 11909296 TI - Topological solitons in nondegenerate one-component chains. AB - The possibility of the existence of topological solitons in one-component chains with a nondegenerate potential of gradient type is proven. The existence and stability of the solitons are ensured by the competing nonlinear nearest-neighbor potential V1 and parabolic second-nearest-neighbor potential V2. Solitonic solutions have been found analytically for piecewise-parabolic V1 and numerically for smoothened nearest-neighbor (NN) potential V(1,delta). Numerical results for the soliton velocity and front width are in good agreement with analytical estimates. The solitons are shown to move at a unique velocity and actually maintain the constant profile as long as the NN potential is smooth enough. The impact of two solitons of different sign is inelastic and leads to their recombination. It is argued that the soliton propagation may constitute an elementary event of structural transformations in the chain. PMID- 11909297 TI - Slow light, induced dispersion, enhanced nonlinearity, and optical solitons in a resonator-array waveguide. AB - We describe an optical transmission line that consists of an array of wavelength scale optical disk resonators coupled to an optical waveguide. Such a structure leads to exotic optical characteristics, including ultraslow group velocities of propagation, enhanced optical nonlinearities, and large dispersion with a controllable magnitude and sign. This device supports soliton propagation, which can be described by a generalized nonlinear Schrodinger equation. PMID- 11909298 TI - Pattern formation via symmetry breaking in nonlinear weakly correlated systems. AB - We study pattern formation initiated by modulation instability in nonlinear partially coherent wave fronts and show that anisotropic noise and/or anisotropic correlation statistics can lead to ordered patterns. PMID- 11909299 TI - Propagation of classical waves in nonperiodic media: scaling properties of an optical Cantor filter. AB - Wave propagation through a subclass of deterministic nonperiodic media, namely, fractal Cantor multilayer structures are investigated theoretically as well as experimentally. Transmission spectra of Cantor structures are found to have two distinctive properties (scalability and sequential splitting) closely related to the geometrical peculiarities of the multilayers. A systematic correlation between structural self-similarity and spectral regularities of Cantor multilayers is established. PMID- 11909300 TI - Numerical studies of left-handed materials and arrays of split ring resonators. AB - We present numerical results on the transmission properties of the left-handed materials (LHMs) and split-ring resonators (SRRs). The simulation results are in qualitative agreement with experiments. The dependence of the transmission through LHMs on the real and imaginary part of the electric permittivity of the metal, the length of the system, and the size of the unit cell are presented. We also study the dependence of the resonance frequency of the array of SRRs on the ring thickness, inner diameter, radial and azimuthal gap, as well as on the electrical permittivity of the board and the embedding medium, where the SRR resides. Qualitatively good agreement with previously published analytical results is obtained. PMID- 11909301 TI - Polarization state of the optical near field. AB - The polarization state of the optical electromagnetic field lying several nanometers above complex dielectric-air interfaces reveals the intricate light matter interaction that occurs in the near-field zone. From the experimental point of view, access to this information is not direct and can only be extracted from an analysis of the polarization state of the detected light. These polarization states can be calculated by different numerical methods, well suited to near-field optics. In this paper, we apply two different techniques (localized Green's function method and differential theory of gratings) to separate each polarization component associated with both electric and magnetic optical near fields produced by nanometer sized objects. A simple dipolar model is used to get an insight into the physical origin of the near-field polarization state. In a second stage, accurate numerical simulations of field maps complete data produced by analytical models. We conclude this study by demonstrating the role played by the near-field polarization in the formation of the local density of states. PMID- 11909302 TI - Lyapunov spectrum from time series using moving boxes. AB - We present a very fast algorithm (few seconds) for estimating full Lyapunov spectrum from time series. The method requires a smaller number of parameters than other time-average algorithms, and is tested for data with different numerical precision, sampling frequencies, total sampling times, and presence of noise. We report conclusive results for the electron density broadband fluctuations of a plasma at the edge of tokamak TBR-1. PMID- 11909304 TI - Kinetic Monte Carlo simulations with minimal searching. AB - Kinetic Monte Carlo (KMC) simulations are used to simulate epitaxial crystal growth. Presently, the fastest reported methods use binary trees to search through a list of rates in O(log(2) M) time, where M is the number of rates. These methods are applicable to an arbitrary set of rates, but typical KMC bond counting schemes involve only a finite set of distinct rates. This allows one to construct a faster list-based algorithm with a computation time that is essentially independent of M. It is found that this algorithm typically reduces computation time by between 30% and 50% for typical simulations, with this factor increasing for larger simulations. PMID- 11909303 TI - Real-space renormalization-group approach to field evolution equations. AB - An operator formalism for the reduction of degrees of freedom in the evolution of discrete partial differential equations (PDE) via real-space renormalization group is introduced, in which cell overlapping is the key concept. Applications to (1+1)-dimensional PDEs are presented for linear and quadratic equations that are first order in time. PMID- 11909305 TI - Efficient method for simulating quantum electron dynamics under the time dependent Kohn-Sham equation. AB - A numerical scheme for solving the time evolution of wave functions under the time-dependent Kohn-Sham (TDKS) equation has been developed. Since the effective Hamiltonian depends on the wave functions, the wave functions and the effective Hamiltonian should evolve consistently with each other. For this purpose, a self consistent loop is required at every time step for solving the time evolution numerically, which is computationally expensive. However, in this paper, we develop a different approach, expressing a formal solution of the TDKS equation, and prove that it is possible to solve the TDKS equation efficiently and accurately by means of a simple numerical scheme without the use of any self consistent loops. PMID- 11909306 TI - Computer simulations of a two-dimensional system with competing interactions. AB - The results and methodology of large scale computer simulations of the two dimensional dipolar Ising model with long-range interactions are reported. Systems as large as 117,649 particles were studied to elucidate the elementary excitations and phase diagram of two-dimensional systems, such as Langmuir monolayers, thin garnet films, and adsorbed films on solid surfaces, which spontaneously form patterns of stripes, bubbles, and intermediately shaped domains. The challenging numerical investigations of large scale systems with long-range interactions at low temperatures were made possible by combining the fast multipole method and a non-Metropolis Monte Carlo sampling technique. Our simulations provide evidence that, at sufficiently high ratios of the repulsive to the attractive coupling constant for the model, twofold stripe order in the systems of interest is lost through a defect-mediated mechanism. Heat capacity data and the excitations observed in our simulations as the system disorders indicate that it is most likely an instance of a Kosterlitz-Thouless phase transition. The results from simulations with and without external field are in excellent agreement with the predictions of an analytic scaling theory [A. D. Stoycheva and S. J. Singer, Phys. Rev. E 64, 016118 (2001)], confirming the phase diagram furnished by the analytic model. The scaling theory suggests that, under certain conditions, defect-mediated stripe melting may be supplanted by Ising like disordering within stripes for small repulsion strength. A qualitative discussion of a model that supports both disordering mechanisms is presented. PMID- 11909307 TI - Improvements on the application of convergence accelerators for the evaluation of some three-electron atomic integrals. AB - Convergence accelerator methods are employed to analyze some of the most difficult three-electron integrals that arise in atomic calculations. These integrals have an explicit dependence on the interelectronic coordinates, and take the form integral r(i)(1)r(j)(2)r(k)(3)r(l)(23)r(m)(31)r(n)(12) exp(( alpha(r1)-beta(r2)-gamma(r3))dr(1)dr(2)dr(3). The focus of the present investigation are the most difficult cases of the parameter set [i, j, k, l, m, n]. Several convergence accelerator techniques are studied, and a comparison presenting the relative effectiveness of each technique is reported. When the convergence accelerator approach is combined with specialized numerical quadrature methods, we find that the overall technique yields high-precision results and is fairly efficient in terms of computational resources. Difficulties associated with the standard numerical precision loss of convergence accelerator techniques are circumvented. PMID- 11909308 TI - Taylor-series expansion and least-squares-based lattice Boltzmann method: Two dimensional formulation and its applications. AB - An explicit lattice Boltzmann method (LBM) is developed in this paper to simulate flows in an arbitrary geometry. The method is based on the standard LBM, Taylor series expansion, and the least-squares approach. The final formulation is an algebraic form and essentially has no limitation on the mesh structure and lattice model. Theoretical analysis for the one-dimensional (1D) case showed that the version of the LBM could recover the Navier-Stokes equations with second order accuracy. A generalized hydrodynamic analysis is conducted to study the wave-number dependence of shear viscosity for the method. Numerical simulations of the 2D lid-driven flow in a square cavity and a polar cavity flow as well as the "no flow" simulation in a square cavity have been carried out. Favorable results were obtained and compared well with available data in the literature, indicating that the present method has good prospects in practical applications. PMID- 11909309 TI - Self-referential method for calculation of the free energy of crystals by Monte Carlo simulation. AB - We propose a Monte Carlo simulation method for the evaluation of free energies in crystalline systems. In principle, the method involves evaluating the free-energy difference between systems of N molecules and 2N molecules. This difference, coupled with the assumption that the free energy is extensive and thus proportional to N, provides sufficient information to obtain the absolute free energy of the crystal. The approach to doubling the system size does not involve insertion or removal of molecules in the system. Instead, the configurations of the molecules are expressed in terms of the normal-mode coordinates of a harmonic lattice. By decoupling certain of these coordinates from the molecule configurations, we obtain a transformation that in effect yields the system-size doubling. The method is examined via application to a system of hard rods in one dimension. This simple model is considered principally because of the availability of an analytic solution for its free energy, which permits accurate testing of the performance and correctness of the proposed method. In using the hard-rod model we also avoid other complications related to treatment of the temperature, and application of normal-mode coordinate decoupling in higher dimensions. The proposed method is shown to be able to provide good results for the free-energy calculation, but further development will be needed before it can be considered practical for general-purpose use. PMID- 11909310 TI - Lattice-switch Monte Carlo method: application to soft potentials. AB - The lattice-switch Monte Carlo method, recently introduced and applied in the context of hard spheres, is extended to particles interacting through a soft potential. The method utilizes a transformation that switches between configurations of two different crystalline structures, allowing the phase space of both structures to be explored in a single simulation and the difference between their free energies to be determined directly. We apply the method to determine the fcc-hcp crystalline phase behavior of the classical Lennard-Jones solid. PMID- 11909311 TI - Thermal treatment of the minority game. AB - We study a cost function for the aggregate behavior of all the agents involved in the minority game (MG) or the bar attendance model (BAM). The cost function allows us to define a deterministic, synchronous dynamic that yields results that have the main relevant features than those of the probabilistic, sequential dynamics used for the MG or the BAM. We define a temperature through a Langevin approach in terms of the fluctuations of the average attendance. We prove that the cost function is an extensive quantity that can play the role of an internal energy of the many-agent system while the temperature so defined is an intensive parameter. We compare the results of the thermal perturbation to the deterministic dynamics and prove that they agree with those obtained with the MG or BAM in the limit of very low temperature. PMID- 11909312 TI - Computer simulation study of irreversible adsorption: coverage fluctuations. AB - In this paper, we develop a cellular automata model to study the coverage fluctuations in monolayers of irreversible adsorbed particles. The effect of bulk diffusion and excluded volume interactions between adsorbed and incoming particles on coverage fluctuations is analyzed by simulations and analytically. We also show that the macroscopic boundary and initial conditions imposed at the system (open or closed cell) determine the effect of these factors on coverage fluctuations. In fact, under certain conditions, the excluded volume interactions only influence fluctuations near the jamming limit. PMID- 11909313 TI - Interband spectrum of weakly coupled stochastic lattice Ginzburg-Landau models. AB - We analyze the excitation spectrum of the generator associated with the relaxation rate to equilibrium in weakly coupled stochastic Ginzburg-Landau models on a spatial lattice Z(d). The spectrum has a quasiparticle interpretation. Depending on d and on the specific interaction, by solving the Bethe-Salpeter equation in the ladder approximation, we show the existence of a stable particle above the upper envelope of the two-particle band, possessing a concave dispersion curve. This result furthers our knowledge about the spectrum of the stochastic dynamics generator. PMID- 11909314 TI - Exact solution for single-scale Gaussian random transport. AB - A quadrature expression is derived for the probability density function of passive tracers advected from a point by a one-dimensional, single-scale, Gaussian velocity field. The effect of trapping on the tracer moments and the Lagrangian velocity variance is explicitly demonstrated. PMID- 11909315 TI - Theory for competing reactions with initially separated components. AB - The asymptotic long-time properties of a system with initially separated components and two competing irreversible reactions A1+B-->C1 and A2+B-->C2 are studied. It is shown that the system is characterized by a single reaction zone, with width growing like t(1/6), in which both reactions occur. Numerical computations of the mean-field kinetic equations confirm these asymptotic results. PMID- 11909316 TI - Energetics of rocked inhomogeneous ratchets. AB - We study the efficiency of frictional thermal ratchets driven by a finite frequency driving force and in contact with a heat bath. The efficiency exhibits varied behavior with driving frequency. Both nonmonotonic and monotonic behavior have been observed. In particular, the magnitude of the efficiency in the finite frequency regime may be more than the efficiency in the adiabatic regime. This is our central result for rocked ratchets. We also show that for the simple potential we have chosen, with only spatial asymmetry (homogeneous system) or only a frictional ratchet (symmetric potential profile), the adiabatic efficiency is always more than in the nonadiabatic case. PMID- 11909317 TI - Fractional Langevin model of memory in financial time series. AB - Financial time series are random with the absolute value of the price index fluctuations having an inverse power-law correlation. A dynamical model of this behavior is proposed using a fractional Langevin equation. The physical basis for this model is the divergence of the microscopic time scale to overlap with the macroscopic time scale: a condition that is not observed in classical statistical mechanics. This time-scale separation provides a mechanism for the market to adjust the volitility of the price index fluctuations. PMID- 11909318 TI - Pernicious effect of physical cutoffs in fractal analysis. AB - Fractal scaling appears ubiquitous, but the typical extension of the scaling range observed is just one to two decades. A recent study has shown that an apparent fractal scaling spanning a similar range can emerge from the randomness in dilute sets. We show that this occurs also in most kinds of nonfractal sets irrespective of defining the fractal dimension by box counting, minimal covering, the Minkowski sausage, Walker's ruler, or the correlation dimension. We trace this to the presence of physical cutoffs, which induce smooth changes in the scaling, and a bias over a couple of decades around some characteristic length. The latter affects also the practical measure of fractality of truly fractal objects. A defensive strategy against artifacts and bias consists in carefully identifying the cutoffs and a quick-and-dirty thumb rule requires to observe fractal scaling over at least three decades. PMID- 11909319 TI - Targeting unknown and unstable periodic orbits. AB - We present a method to target and subsequently control (if necessary) orbits of specified period but otherwise unknown stability and position. For complex systems where the dynamics is often mixed [e.g., coexistence of regular and chaotic regions in area-preserving (Hamiltonian) systems], this targeting algorithm offers a way to not only gently bring the system from the chaotic domain to an unstable periodic orbit (where control is applied), but also to access stable regions of phase space (where control is not necessary) from within the stochastic regions. The technique is quite general and applies equally well to dissipative or conservative discrete maps and continuous flows. PMID- 11909320 TI - Low-dimensional chaos in zero-Prandtl-number Benard-Marangoni convection. AB - Three-dimensional surface-tension-driven Benard convection at zero Prandtl number is computed in the smallest possible doubly periodic rectangular domain that is compatible with the hexagonal flow structure at the linear stability threshold of the quiescent state. Upon increasing the Marangoni number beyond this threshold, the initially stationary flow becomes quickly time dependent. We investigate the transition to chaos for the case of a free-slip bottom wall by means of an analysis of the kinetic energy time series. We observe a period-doubling scenario for the transition to chaos of the energy attractor, intermittent behavior of a component of the mean velocity field, three characteristic energy levels, and two frequencies that contain a considerable amount of the power spectral density connected with the kinetic energy time series. PMID- 11909321 TI - Density relaxation of a near-critical fluid in response to local heating and low frequency vibration in microgravity. AB - The response of a confined near-critical fluid to local heating in the presence of vibration is studied by means of two-dimensional numerical simulations of the compressible and unsteady Navier-Stokes equations written for a van der Waals fluid. As in the experiments performed two years ago onboard the Mir orbital station, two different regimes of density distribution are observed. For sufficiently low frequency and high amplitude vibration, two thermal plumes develop from the heat source along the vibration axis. Otherwise (higher frequency and/or lower amplitude), density inhomogeneities caused by heating stay around the heat source. For this regime, the pair of vortices created in each half period absorbs the preceding one, while it is convected away for the double plume regime. As time goes on, this process repeats, with a lateral extension of the low density region. At lower frequencies, instabilities appear in the flow, thus corroborating again microgravity experiments. PMID- 11909322 TI - Spiral-defect chaos: Swift-Hohenberg model versus Boussinesq equations. AB - Spiral-defect chaos (SDC) in Rayleigh-Benard convection is a well-established spatio-temporal complex pattern, which competes with stationary rolls near the onset of convection. The characteristic properties of SDC are accurately described on the basis of the standard three-dimensional Boussinesq equations. As a much simpler and attractive two-dimensional model for SDC generalized Swift Hohenberg (SH) equations have been extensively used in the literature from the early beginning. Here, we show that the description of SDC by SH models has to be considered with care, especially regarding its long-time dynamics. For parameters used in previous SH simulations, SDC occurs only as a transient in contrast to the experiments and the rigorous solutions of the Boussinesq equations. The small scale structure of the vorticity field at the spiral cores, which might be crucial for persistent SDC, is presumably not perfectly captured in the SH model. PMID- 11909323 TI - Fluid motions in the Earth's core inferred from time spectral features of the geomagnetic field. AB - The aim of this work is to investigate the time spectral features of the main geomagnetic field fluctuations as measured on the Earth's surface in connection with a nontraditional turbulent dynamics of the fluid motions in the outer layers of the Earth's liquid core. The average geomagnetic field spectrum is found to be a power law, characterized by a spectral exponent alpha approximately -11/3, on time scales longer than 5 yr. We discuss the spectral exponent in connection with an intense magnetic field in the Earth's core and with a vortex coalescence process in a regime of drift-wave turbulence. PMID- 11909324 TI - Collisionless damping of nonlinear dust ion acoustic wave due to dust charge fluctuation. AB - A dissipation mechanism for the damping of the nonlinear dust ion acoustic wave in a collisionless dusty plasma consisting of nonthermal electrons, ions, and variable charge dust grains has been investigated. It is shown that the collisionless damping due to dust charge fluctuation causes the nonlinear dust ion acoustic wave propagation to be described by the damped Korteweg-de Vries equation. Due to the presence of nonthermal electrons, the dust ion acoustic wave admits both positive and negative potential and it suffers less damping than the dust acoustic wave, which admits only negative potential. PMID- 11909325 TI - Electron anisotropic scattering in gases: a formula for Monte Carlo simulations. AB - The purpose of this Brief Report is to point out the mistake in a formula for anisotropic electron scattering, previously published in Phys. Rev. A 41, 1112 (1990), which is widely used in Monte Carlo models of gas discharges. Anisotropic electron scattering is investigated based on the screened Coulomb potential between electrons and neutral atoms. The approach is also applied for electron scattering by nonpolar neutral molecules. Differential cross sections for electron scattering by Ar, N2, and CH4 are constructed on the basis of momentum and integrated cross sections. The formula derived in this paper is useful for Monte Carlo simulations of gas discharges. PMID- 11909326 TI - Simple stochastic model for optical tunneling. AB - A simple model, derived from a Brownian-motion scheme, is capable of interpreting the results of delay-time measurements relative to frustrated total reflection experiments at the microwave scale but also in the visible region. In this framework we also obtain a plausible description of the trajectories (rays) inside the tunneling region, the air gap between two paraffin prisms. PMID- 11909327 TI - Fast algorithm for generating long self-affine profiles. AB - We introduce a fast algorithm for generating long self-affine profiles. The algorithm, which is based on the fast wavelet transform, is faster than the conventional Fourier filtering algorithm. In addition to increased performance for large systems, the algorithm, named the wavelet filtering algorithm, a priori gives rise to profiles for which the long-range correlation extends throughout the entire system independently of the length scale. PMID- 11909328 TI - Calculation of the Coulomb energy in quasi-two-dimensional systems. AB - In this paper we obtain expressions for the Coulomb energy in infinite quasi-two dimensional systems in the form of fast converging Ewald sums in the three dimensional coordinate and two-dimensional inverse space needed for accurate formulation of periodic boundary conditions in computer simulations. Numerical tests evidence that the acceptable accuracy in the total energy is achieved by taking into account rather small number of terms in the sum for the inverse space responsible for the long-range part of interaction. PMID- 11909329 TI - Comment on "Symmetric path integrals for stochastic equations with multiplicative noise". AB - We recall our approach through discretizations for path integrals and its general results for representations of probability densities. It is shown that the result of Arnold [P. Arnold, Phys. Rev. E 61, 6099 (2000)] is a particular case of our work. PMID- 11909330 TI - Using novel variable transformations to enhance conformational sampling in molecular dynamics. AB - One of the computational "grand challenges" is to develop methodology capable of sampling conformational equilibria in systems with rough energy landscapes. Here, a significant step forward is made by combining molecular dynamics with a novel variable transformation designed to enhance sampling by reducing barriers without introducing bias and, thus, to preserve, perfectly, equilibrium properties. PMID- 11909331 TI - Multimode interferometer for guided matter waves. AB - Atoms can be trapped and guided with electromagnetic fields, using nanofabricated structures. We describe the fundamental features of an interferometer for guided matter waves, built of two combined Y-shaped beam splitters. We find that such a device is expected to exhibit high contrast fringes even in a multimode regime, analogous to a white light interferometer. PMID- 11909332 TI - Localized single-photon wave functions in free space. AB - We solve the joint open problems of photon localization and single-photon wave functions in the context of spontaneous emission from an excited atom in free space. Our wave functions are well-defined members of a discrete orthonormal function set. Both the degree and shape of the localization are controlled by entanglement mapping onto the atom wave function, even though the atom is remote from the photon. PMID- 11909333 TI - Observable Dirac-type singularities in Berry's phase and the monopole. AB - A three-dimensional generalization of the sign-change ( pi phase shift) rule for adiabatic cycles of spin-1/2 or two-state wave functions encircling a degeneracy in the parameter space of the Hamiltonian yields a Dirac-type singularity wherein any closed circuit of the adiabatic cycle in which the degeneracy is "looped" results in an observable +/-2pi phase shift. It is concluded that an interferometer loop similarly taken around a magnetic monopole of strength n/2 yields an observable +/-2npi phase shift, n being an integer. PMID- 11909334 TI - Matter-wave interferometer for large molecules. AB - We demonstrate a near-field Talbot-Lau interferometer for C70 fullerene molecules. Such interferometers are particularly suitable for larger masses. Using three free-standing gold gratings of 1 microm period and a transversally incoherent but velocity-selected molecular beam, we achieve an interference fringe visibility of 40% with high count rate. Both the high visibility and its velocity dependence are in good agreement with a quantum simulation that takes into account the van der Waals interaction of the molecules with the gratings and are in striking contrast to a classical moire model. PMID- 11909335 TI - A rotating black ring solution in five dimensions. AB - The vacuum Einstein equations in five dimensions are shown to admit a solution describing a stationary asymptotically flat spacetime regular on and outside an event horizon of topology S1xS2. It describes a rotating "black ring." This is the first example of a stationary asymptotically flat vacuum solution with an event horizon of nonspherical topology. The existence of this solution implies that the uniqueness theorems valid in four dimensions do not have simple five dimensional generalizations. It is suggested that increasing the spin of a spherical black hole beyond a critical value results in a transition to a black ring, which can have an arbitrarily large angular momentum for a given mass. PMID- 11909336 TI - A uniqueness theorem for the anti-de Sitter soliton. AB - The stability of physical systems depends on the existence of a state of least energy. In gravity, this is guaranteed by the positive energy theorem. For topological reasons, this fails for nonsupersymmetric Kaluza-Klein compactifications, which can decay to arbitrarily negative energy. For related reasons, this also fails for the anti-de Sitter (AdS) soliton, a globally static, asymptotically toroidal Lambda<0 spacetime with negative mass. Nonetheless, arguing from the AdS conformal field theory (AdS/CFT) correspondence, Horowitz and Myers proposed a new positive energy conjecture, which asserts that the AdS soliton is the unique state of least energy in its asymptotic class. We give a new structure theorem for static Lambda<0 spacetimes and use it to prove uniqueness of the AdS soliton. Our results offer significant support for the new positive energy conjecture and add to the body of rigorous results inspired by the AdS/CFT correspondence. PMID- 11909337 TI - Hall-drift induced magnetic field instability in neutron stars. AB - In the presence of a strong magnetic field and under conditions as realized in the crust and the superfluid core of neutron stars, the Hall drift dominates the field evolution. We show by a linear analysis that, for a sufficiently strong large-scale background field depending at least quadratically on position in a plane conducting slab, an instability occurs which rapidly generates small-scale fields. Their growth rates depend on the choice of the boundary conditions, increase with the background field strength, and may reach 10(3) times the Ohmic decay rate. The effect of that instability on the rotational and thermal evolution of neutron stars is discussed. PMID- 11909338 TI - Gravothermal collapse of self-interacting dark matter halos and the origin of massive black holes. AB - Black hole formation is an inevitable consequence of relativistic core collapse following the gravothermal catastrophe in self-interacting dark matter (SIDM) halos. Very massive SIDM halos form supermassive black holes (SMBHs) > or about 10(6)M(middle dot in circle) directly. Smaller halos believed to form by redshift z = 5 produce seed black holes of (10(2)-10(3))M(middle dot in circle) which can merge and/or accrete to reach the observational SMBH range. This scenario for SMBH formation requires no baryons, no prior star formation, and no other black hole seed mechanism. PMID- 11909339 TI - Generalized symmetry breaking on orbifolds. AB - We reconsider the phenomenon of mass generation via coordinate-dependent compactifications of higher-dimensional theories on orbifolds. For definiteness, we study a generic five-dimensional theory compactified on S(1)/Z(2). We show that the presence of fixed points, where the fields or their derivatives may be discontinuous, permits new realizations of the Scherk-Schwarz mechanism where, for example, the mass terms are localized at the orbifold fixed points. Our technique can be used to describe the explicit breaking of global flavor symmetries and supersymmetries by brane-localized mass terms. It can also be applied to the spontaneous breaking of local symmetries, such as gauge symmetries or supergravities. PMID- 11909340 TI - Spontaneously broken spacetime symmetries and Goldstone's theorem. AB - Goldstone's theorem states that there is a massless mode for each broken symmetry generator. It has been known for a long time that the naive generalization of this counting fails to give the correct number of massless modes for spontaneously broken spacetime symmetries. We explain how to get the right count of massless modes in the general case, and discuss examples involving spontaneously broken Poincare and conformal invariance. PMID- 11909341 TI - Demonstration of the lateral casimir force. AB - The lateral Casimir force between a sinusoidally corrugated gold coated plate and large sphere was measured for surface separations between 0.2 to 0.3 microm using an atomic force microscope. The measured force shows the required periodicity corresponding to the corrugations. It also exhibits the necessary inverse fourth power distance dependence. The obtained results are shown to be in good agreement with a complete theory taking into account the imperfectness of the boundary metal. This demonstration opens new opportunities for the use of the Casimir effect for lateral translation in microelectromechanical systems. PMID- 11909344 TI - How sensitive is a neutrino factory to the angle theta(13)? AB - We consider the impact of nonstandard interactions (NSI) of neutrinos on the determination of neutrino mixing parameters at a neutrino factory using nu(-)(e)- >nu(-)(mu) "golden channels" for the measurement of theta(13). We show how a small residual NSI leads to a drastic loss in sensitivity in theta(13), of up to 2 orders of magnitude. This can be somewhat overcome if two baselines are combined. PMID- 11909342 TI - Measurements of the cross section for e(+)e(-) --> hadrons at center-of-mass energies from 2 to 5 GeV. AB - We report values of R = sigma(e(+)e(-)-->hadrons)/sigma(e(+)e(-)-->mu(+)mu(-)) for 85 center-of-mass energies between 2 and 5 GeV measured with the upgraded Beijing Spectrometer at the Beijing Electron-Positron Collider. PMID- 11909343 TI - Observation of exclusive B --> D(*)K(*-) decays. AB - We report the first observation of the exclusive decays B-->D((*))K(*-), using 9.66 x 10(6) BB pairs collected at the Upsilon(4S) with the CLEO detector. We measure the following branching fractions: B(B--->D(0)K(*-)) = (6.1+/-1.6+/ 1.7)x10(-4), B(B(0)-->D(+)K(*-)) = (3.7+/-1.5+/-1.0)x10(-4), B(B(0)-->D(*+)K(*-)) = (3.8+/-1.3+/-0.8)x10(-4), and B(B--->D(*0)K(*-)) = (7.7+/-2.2+/-2.6)x10(-4). The B-->D(*)K(*-) branching ratios are the averages of those corresponding to the 00 and 11 helicity states. The errors shown are statistical and systematic, respectively. PMID- 11909347 TI - Masses of the 70(-) baryons in large N(c) QCD. AB - The masses of the negative parity 70-plet baryons are analyzed in large N(c) QCD to order 1/N(c) and to first order in SU(3) symmetry breaking. The existing experimental data are well reproduced and 20 new observables are predicted. The leading order SU(6) spin-flavor symmetry breaking is small and, as it occurs in the quark model, the subleading in 1/N(c) hyperfine interaction is the dominant source of the breaking. It is found that the Lambda(1405) and Lambda(1520) are well described as three-quark states and spin-orbit partners. New relations between splittings in different SU(3) multiplets are found. PMID- 11909346 TI - Towards collinear evolution equations in electroweak theory. AB - We investigate collinear factorization in electroweak radiative corrections to hard inclusive processes at the TeV scale. Because of the uncanceled double logs found previously, we find a factorization pattern which is qualitatively different from the analogous one in QCD. New types of splitting functions emerge which are needed to describe the initial beam charges and are dependent on an infrared cutoff provided by the symmetry breaking scale. We derive such splitting functions at the one-loop level for broken SU(2) gauge theory, and we also discuss the structure functions' evolution equations, under the assumption that isospin breaking terms are sufficiently subleading at higher orders. PMID- 11909348 TI - Longitudinal flow of protons from (2-8)A GeV central Au+Au collisions. AB - Rapidity distributions of protons from central 197Au+197Au collisions measured by the E895 Collaboration in the energy range from (2-8)A GeV at the Brookhaven AGS are presented. Longitudinal flow parameters derived using a thermal model including collective longitudinal expansion are extracted from these distributions. The results show an approximately linear increase in the longitudinal flow velocity, (L), as a function of the logarithm of beam energy. PMID- 11909350 TI - Evidence for a new low-lying resonance state in 7He. AB - Low-lying resonance states in 7He(6He+n), formed after fragmentation reactions of a 227 MeV/nucleon 8He beam on a carbon target, have been studied. Coincidences between 6He nuclei and neutrons, corresponding to the one-neutron knockout channel in 8He, were selected. The relative energy spectrum in the 6He+n system shows a structure, which is interpreted as the 7He ( Ipi = 3/2(-)) ground state, unbound with 0.43(2) MeV relative to the 6He+n system and a width of Gamma = 0.15(8) MeV overlapping with an excited ( Ipi = 1/2(-)) state observed at 1.0(1) MeV with a width of Gamma = 0.75(8) MeV. PMID- 11909349 TI - Spin-momentum correlations in quasielastic electron scattering from deuterium. AB - The spin-momentum correlation parameter A(V)(ed) was measured for the 2H-->(e- >,e'p)n reaction for missing momenta up to 350 MeV/c at Q2 = 0.21 (GeV/c)(2) for quasielastic scattering of polarized electrons from vector-polarized deuterium. The data give detailed information about the deuteron spin structure and are in good agreement with the results of microscopic calculations based on realistic nucleon-nucleon potentials and including various spin-dependent reaction mechanism effects. The experiment reveals in a most direct manner the effects of the D state in the deuteron ground-state wave function and shows the importance of isobar configurations for this reaction. PMID- 11909352 TI - Measurement of absolute cross sections for excitation of the 3s(2)3p(5) 2P(o)(3/2)- 3s(2)3p(5) 2P(o)(1/2) fine-structure transition in Fe9+. AB - Experimental cross sections are reported for the 3s(2)3p(5) 2P(o)(3/2)- 3s(2)3p(5) 2P(o)(1/2) transition in Fe9+ located at 1.945 eV. The center-of-mass interaction energies are in the range of 1.72 eV (below threshold) through threshold, to 5.6 eV (2.9 x threshold). Data are compared with results of a 49 state Breit-Pauli R-matrix theory. The experiment detects structures at 3.5 and 4.6 eV corresponding to enhancement of the direct excitation via many narrow, closely spaced resonances about these energies calculated by the theory. Iron is present in practically every astrophysical object, as well as being an impurity in fusion plasmas. Present data are the first electron-energy-loss measurements on a highly charged iron ion. PMID- 11909351 TI - Quadrupole moment of the 11(-) intruder isomer in 196Pb and its implications for the 16(-) shears band head. AB - The quadrupole moment of the 11(-) isomer in 196Pb has been measured by the level mixing spectroscopy method. This state has a pi(3s(-2)(1/2)1h(9/2)1i(13/2))11(-) configuration which is involved in most of the shears band heads in the Pb region. The first directly measured value of Q(s)(11(-)) = (-)3.41(66) b, coupled to the previously known quadrupole moment of the nu(1i(-2)(13/2))12(+) isomer allows us to estimate the quadrupole moment of the 16(-) shears band head as Q(s)(16(-)) = -0.32(10) b. The experimental values are compared to tilted axis cranking calculations, giving insight into the validity of the additivity approach to couple quadrupole moments and on the amount of deformation in the shears bands. PMID- 11909353 TI - Simultaneous projectile-target ionization: a novel approach to (e, 2e) experiments on ions. AB - A kinematically complete experiment for simultaneous ionization of a projectile and target has been performed for 3.6 MeV/u C2+ on He collisions measuring the final vector momenta of the He1+ recoil ion and of two electrons (projectile, target) in coincidence with the emerging C3+ projectile. The feasibility of an event-by-event separation of the various reaction channels, among them the ionization of C2+ by the interaction with a quasifree target electron, is demonstrated in agreement with six-body classical trajectory Monte Carlo calculations, paving the way to kinematically complete electron-ion scattering experiments. PMID- 11909354 TI - Transporting and time reversing light via atomic coherence. AB - We study basic issues central to the storage of quantum information in a coherently prepared atomic medium such as the role of adiabaticity. We also propose and demonstrate transporting, multiplexing, and time reversing of stored light. PMID- 11909355 TI - Generation of single dispersion precompensated 1-fs pulses by shaped-pulse optimized high-order stimulated Raman scattering. AB - We propose and theoretically analyze a new approach for generating and shaping 1 fs pulses. It combines the ideas of strong-field molecular optics and optimal control to manipulate light generation in a pump-probe Raman regime. Flexible phase control over the generated spectrum of about 3 eV width is achieved by controlling the input pulses and maximizing the coherence of medium excitation by adiabatically aligning molecules in the medium with a specially shaped pump pulse. The generated pulse is optimized for an output window, precompensating for its dispersion to all orders. PMID- 11909356 TI - Observed scattering into a dark optical vortex core. AB - The dark core of an optical vortex was used to detect on-axis, forward-scattered light from a colloidal solution in the single and multiple scattering regimes. Using no adjustable parameters we obtain good agreement with a concentration dependent scattering model. PMID- 11909358 TI - Ultrasound tunneling through 3D phononic crystals. AB - We report the study of ultrasound tunneling in 3D phononic crystals, consisting of fcc arrays of close-packed tungsten carbide beads in water. The transmission coefficient, phase velocity, and group velocity were measured along the [111] direction, allowing us to systematically investigate the tunneling of ultrasound at frequencies in the lowest band gap. Our experimental data are interpreted using multiple scattering theory, which provides a good explanation of our results. The effect of absorption and the difference between the tunneling of classical waves and quantum waves are discussed. PMID- 11909359 TI - New effect in near-field thermal emission. AB - A near-field effect has been discovered experimentally in thermal radio emission of an absorbing dielectric medium. It is related to a specific character of the distribution of a quasistationary field component near a radiating surface. The effect consists of the fact that the effective depth of the received emission formation appears to be less than the skin-layer depth and depends on the size of the receiver antenna and its height above the surface. It can be considered as a new source of information about depth temperature distribution. A theory has been developed that allows for determining the relative contribution of wave and quasistationary components to the Plank emission received near the surface. PMID- 11909357 TI - Leading off-diagonal correction to the form factor of large graphs. AB - Using periodic-orbit theory beyond the diagonal approximation we investigate the form factor, K(tau), of a generic quantum graph with mixing classical dynamics and time-reversal symmetry. We calculate the contribution from pairs of self intersecting orbits that differ from each other only in the orientation of a single loop. In the limit of large graphs, these pairs produce a contribution 2tau(2) to the form factor which agrees with random-matrix theory. PMID- 11909360 TI - Novel method for multiturn extraction: trapping charged particles in islands of phase space. AB - A novel method for multiturn extraction from a circular particle accelerator is presented, based on trapping particles into islands of phase space generated by nonlinear resonances. By appropriate use of sextupoles and octupoles, stable islands can be created at small amplitude in phase space. By varying the linear tune, particles can be trapped inside these islands and then transported towards higher amplitudes for extraction. Results of numerical simulations are discussed. PMID- 11909362 TI - Control of neoclassical tearing modes by sawtooth control. AB - The onset of a neoclassical tearing mode (NTM) depends on the existence of a large enough seed island. It is shown in the Joint European Torus that NTMs can be readily destabilized by long-period sawteeth, such as obtained by sawtooth stabilization from ion-cyclotron heating or current drive. This has important implications for burning plasma scenarios, as alpha particles strongly stabilize the sawteeth. It is also shown that, by adding heating and current drive just outside the inversion radius, sawteeth are destabilized, resulting in shorter sawtooth periods and larger beta values being obtained without NTMs. PMID- 11909363 TI - Alpha-tail production with ion-cyclotron-resonance heating of 4He-beam ions in JET plasmas. AB - Third-harmonic ion-cyclotron-resonance heating of 4He-beam ions has produced for the first time on the JET tokamak high-energy populations of 4He ions to simulate 3.5 MeV fusion-born alpha (alpha) particles. Acceleration of 4He ions to the MeV energy range is confirmed by gamma-ray emission from the nuclear reaction 9Be(alpha,ngamma) 12C and excitation of Alfven eigenmodes. Concomitant electron heating and sawtooth stabilization are observed. The scheme could be used in next step tokamaks to gain information on trapped alpha particles and to test alpha diagnostics in the early nonactivated phase of operation. PMID- 11909364 TI - Measurements of nonlinear growth of ion-acoustic waves in two-ion-species plasmas with thomson scattering. AB - We report the first Thomson-scattering measurements of the growth of ion-acoustic waves in well-characterized multi-ion-species plasmas consisting of gold and beryllium. We observe that only the berylliumlike mode grows, verifying linear kinetic theory. In addition, a twofold increase in ion temperature is measured when ion-acoustic waves are excited to large amplitudes by stimulated Brillouin scattering (SBS). This increase in ion temperature is a strong indication of hot ions due to trapping. We explain the measured SBS reflectivity by nonlinear detuning of the SBS instability due to these trapping effects. PMID- 11909365 TI - Effects of circulating energetic ions on sawtooth oscillations. AB - In contrast to the well-known result that the effects of the trapped energetic ions (TEI) on the internal kink mode are due to the toroidal precession of the TEI, it is found that the effects of the circulating energetic ions (CEI) on sawtooth are due to the toroidal circulation of the CEI. The effects of the CEI on sawtooth oscillations are found to be different from the well-known purely stabilizing effects of the TEI on sawtooth oscillations; the toroidal circulation of the co-CEI provides an additional sink of free energy and stabilizes the mode; the toroidal circulation of the counter-CEI provides an additional source of free energy and destabilizes the mode. PMID- 11909361 TI - Generation of GW radiation pulses from a VUV free-electron laser operating in the femtosecond regime. AB - Experimental results are presented from vacuum-ultraviolet free-electron laser (FEL) operating in the self-amplified spontaneous emission (SASE) mode. The generation of ultrashort radiation pulses became possible due to specific tailoring of the bunch charge distribution. A complete characterization of the linear and nonlinear modes of the SASE FEL operation was performed. At saturation the FEL produces ultrashort pulses (30-100 fs FWHM) with a peak radiation power in the GW level and with full transverse coherence. The wavelength was tuned in the range of 95-105 nm. PMID- 11909366 TI - From current-driven to neoclassically driven tearing modes. AB - In the TCV tokamak, the m/n = 2/1 island is observed in low-density discharges with central electron-cyclotron current drive. The evolution of its width has two distinct growth phases, one of which can be linked to a "conventional" tearing mode driven unstable by the current profile and the other to a neoclassical tearing mode driven by a perturbation of the bootstrap current. The TCV results provide the first clear observation of such a destabilization mechanism and reconcile the theory of conventional and neoclassical tearing modes, which differ only in the dominant driving term. PMID- 11909367 TI - Coherent optical phonon oscillations in bulk GaN excited by far below the band gap photons. AB - We report on generation of coherent optical phonon oscillations in 150 microm thick bulk GaN. With photon energy far below the band gap, the generation mechanisms of coherent phonon modes of A1(LO), high- and low-frequency E2 are revealed to be the impulsive stimulated Raman scattering. We find that one among the two degenerate E2 modes is selectively detected with a proper choice of probe polarization. Dephasing times range from 1.5 to 70 ps for different modes, and phonon-three-photon absorbed carrier interactions are compared between the A1(LO) and the E2 mode. PMID- 11909345 TI - Measurement of B --> K*gamma branching fractions and charge asymmetries. AB - The branching fractions of the exclusive decays B0-->K(*0)gamma and B+- >K(*+)gamma are measured from a sample of (22.74+/-0.36)x10(6) BB decays collected with the BABAR detector at the PEP-II asymmetric e(+)e(-) collider. We find B (B0-->K(*0)gamma) = [4.23+/-0.40(stat)+/-0.22(syst)]x10(-5), B(B+- >K(*+)gamma) = [3.83+/-0.62(stat)+/-0.22(syst)]x10(-5) and constrain the CP violating charge asymmetry to be -0.170K(*)gamma)<0.082 at 90% C.L. PMID- 11909368 TI - Hot nanotubes: stable heating of individual multiwall carbon nanotubes to 2000 k induced by the field-emission current. AB - Field emission (FE) electron spectroscopy from an individual multiwalled carbon nanotube (MWNT) is used to measure quantitatively stable temperatures at the apex, T(A), of up to 2000 K induced by FE currents approximately 1 microA. The high T(A) is due to Joule heating along the length of the MWNT. These measurements also give directly the resistance of the individual MWNT which is shown to decrease with temperature, and explain the phenomenon of FE-induced light emission which was observed simultaneously. The heating permits thermal desorption of the MWNT and, hence, excellent current stability. PMID- 11909369 TI - A novel Monte Carlo scheme for the rapid equilibration of atomistic model polymer systems of precisely defined molecular architecture. AB - Two novel connectivity-altering atomistic Monte Carlo moves are presented for the fast equilibration of condensed phases of long-chain systems with a variety of chain architectures. With the new moves, isotropic or oriented melts of linear or long-chain branched polymers, dense brushes of terminally grafted macromolecules, and cyclic peptides can be simulated. Results concerning the structural, conformational, and volumetric properties of linear, monodisperse polyethylene melts, simulated with a new united-atom molecular model, are in excellent agreement with experimental data. PMID- 11909370 TI - Hydrodynamics of topological defects in nematic liquid crystals. AB - We show that backflow, the coupling between the order parameter and the velocity fields, has a significant effect on the motion of defects in nematic liquid crystals. In particular, the defect speed can depend strongly on the topological strength in two dimensions and on the sense of rotation of the director about the core in three dimensions. PMID- 11909371 TI - Fractal geometry of surface areas of sand grains probed by pulsed field gradient NMR. AB - Pulsed field gradient NMR self-diffusion studies of water were used to determine surface-to-volume ratios and specific surface areas of the grains forming a glacial sand deposit. Both quantities exhibit a noninteger power-law dependence as a function of the diameters of the grains. The associated fractal dimensions of the surface area ( D(s)) and of the pore volume ( D(v)) are found to be D(s) D(v) = -0.70+/-0.05 and D(s) = 2.20+/-0.05. The results demonstrate that NMR studies with native pore fluids are suitable to investigate the fractal nature of natural, unconsolidated porous materials. PMID- 11909372 TI - Formation of vacancy-impurity complexes by kinetic processes in highly As-doped Si. AB - Positron annihilation experiments have been applied to verify the formation mechanism of electrically inactive vacancy-impurity clusters in highly n-type Si. We show that the migration of V-As pairs at 450 K leads to the formation of V-As2 complexes, which in turn convert to stable V-As3 defects at 700 K. These processes manifest the formation of V-As3 as the dominant vacancy-impurity cluster in highly n-type Si. They further explain the electrical deactivation and clustering of As in epitaxial or ion-implanted Si during postgrowth heat treatment at 700 K. PMID- 11909373 TI - Key to understanding interstitial H2 in Si. AB - A new IR absorption line at 3191.1 cm(-1) has been discovered for the interstitial HD molecule in Si. This new line appears for sample temperatures above approximately 20 K and lies 73.9 cm(-1) below the 3265.0 cm(-1) line previously observed for HD. We attribute the 73.9 cm(-1) energy difference to the rotation of the interstitial HD molecule. The selection rules associated with these two lines are consistent with the puzzling absence of an ortho-para splitting in the IR absorption spectra of H2 and D2 in Si. PMID- 11909374 TI - X-ray photoelectron spectroscopy study of disordering in Gd2(Ti1-xZrx)2O7 pyrochlores. AB - The dramatic increases in ionic conductivity in Gd2(Ti1-xZrx)2O7 solid solution are related to disordering on the cation and anion lattices. Disordering in Gd2(Ti1-xZrx)2O7 was characterized using x-ray photoelectron spectroscopy (XPS). As Zr substitutes for Ti in Gd2Ti2O7 to form Gd2(Ti1-xZrx)2O7 (0.25 < x < or =0.75), the corresponding O 1s XPS spectrum merges into a single symmetric peak. This confirms that the cation antisite disorder occurs simultaneously with anion disorder. Furthermore, the O 1s XPS spectrum of Gd2Zr2O7 experimentally suggests the formation of a split vacancy. PMID- 11909375 TI - Behavior of partial monolayers of argon adlayers deposited on graphite. AB - This calculation examines the properties of partial monolayers of argon deposited on graphite, over an interval 20< or =T< or =90 K. In particular, we focus upon the phase transitions. Two peaks in the specific heat are found. The first peak is very sharp and narrow. It has been identified as a rotation of the adlayer from an angle off the substrate symmetry axis, to that axis. The calculated orientational behavior is compared with predictions of the two dimensional Ising model. The broad peak around 49.5 K is identified as a melting transition which is characterized by a loss of local sixfold symmetry. PMID- 11909376 TI - Limits of the hydrodynamic no-slip boundary condition. AB - A controversial point in fluid dynamics is to distinguish the relative importance of surface roughness and fluid-surface intermolecular interactions in determining the boundary condition. Here hydrodynamic forces were compared for flow of Newtonian fluids past surfaces of variable roughness but similar, poorly wetted, surface chemistry. The critical shear stress and shear rate to observe deviations from predictions using the no-slip boundary condition increased nearly exponentially with increasing roughness and diverged at approximately 6 nm rms roughness. We conclude that local intermolecular interactions dominated when the surface was very smooth, but roughness dominated otherwise. This quantifies the limits of both ideas. PMID- 11909377 TI - Phase separation due to quantum mechanical correlations. AB - Can phase separation be induced by strong electron correlations? We present a theorem that affirmatively answers this question in the Falicov-Kimball model away from half filling, for any dimension. In the ground state the itinerant electrons are spatially separated from the classical particles. PMID- 11909378 TI - Josephson current in Luttinger liquid-superconductor junctions. AB - We study the Josephson current through a Luttinger liquid in contact with two superconductors. We show that it can be deduced from the Casimir energy in a two boundary version of the sine-Gordon model. We develop a new thermodynamic Bethe ansatz, which, combined with a subtle analytic continuation procedure, allows us to calculate this energy in closed form, and obtain the complete current crossover function from the case of complete normal to complete Andreev reflection. PMID- 11909379 TI - Coupling between spin and orbital degrees of freedom in KCuF3. AB - We present the results of resonant x-ray scattering experiments on KCuF3. Structurally forbidden reflections, corresponding to magnetic and 3d-orbital long range order, have been observed. Integrated intensities have been measured as a function of incident energy, polarization, azimuthal angle, and temperature. The results give evidence for a strong coupling between orbital and spin degrees of freedom. The interplay between magnetic and orbital order parameters is revealed by the temperature dependence of the intensity of orbital Bragg peaks. PMID- 11909380 TI - Strong enhancement of drag and dissipation at the weak- to strong-coupling phase transition in a bilayer system at a total Landau level filling nu = 1. AB - We consider a bilayer electronic system at a total Landau level filling factor nu = 1, and focus on the transition from the regime of weak interlayer coupling to that of the strongly coupled (1,1,1) phase (or "quantum Hall ferromagnet"). Making the assumption that in the transition region the system is made of puddles of the (1,1,1) phase embedded in a bulk of the weakly coupled state, we show that the transition is accompanied by a strong increase in longitudinal Coulomb drag that reaches a maximum of approximately h/2e(2). In that regime the longitudinal drag increases with decreasing temperature. PMID- 11909382 TI - Nonmonotonic d(x(2)-y(2)) superconducting order parameter in Nd2-xCexCuO4. AB - Low energy polarized electronic Raman scattering of the electron-doped superconductor Nd2-x Ce x CuO4 ( x = 0.15, T(c) = 22 K) has revealed a nonmonotonic d(x(2)-y(2)) superconducting order parameter. It has a maximum gap of 4.4k(B)T(c) at Fermi surface intersections with an antiferromagnetic Brillouin zone (the "hot spots") and a smaller gap of 3.3k(B)T(c) at fermionic Brillouin zone boundaries. The gap enhancement in the vicinity of the hot spots emphasizes the role of antiferromagnetic fluctuations and the similarity in the origin of superconductivity for electron- and hole-doped cuprates. PMID- 11909381 TI - Electronic structure of the trilayer cuprate superconductor Bi(2)Sr(2)Ca(2)Cu(3)O(10+delta). AB - The low-energy electronic structure of the nearly optimally doped trilayer cuprate superconductor Bi(2)Sr(2)Ca(2)Cu(3)O(10+delta) is investigated by angle resolved photoemission spectroscopy. The normal state quasiparticle dispersion and Fermi surface and the superconducting d-wave gap and coherence peak are observed and compared with those of single- and bilayer systems. We find that both the superconducting gap magnitude and the relative coherence-peak intensity scale linearly with T(c) for various optimally doped materials. PMID- 11909384 TI - Inhomogeneous magnetism in single crystalline Sr(3)CuIrO(6+delta): implications to phase-separation concepts. AB - The single crystalline form of an insulator, Sr(3)CuIrO(6+delta), is shown to exhibit unexpectedly more than one magnetic transition (at 5 and 19 K) with spin glass-like magnetic susceptibility behavior. On the basis of this finding, viz., inhomogeneous magnetism in a chemically homogeneous material, we propose that the idea of "phase separation" described for manganites is more widespread in different ways. The observed experimental features enable us to make a comparison with the predictions of a recent toy model on magnetic phase separation in an insulating environment. PMID- 11909383 TI - Band structure and density of States effects in co-based magnetic tunnel junctions. AB - Utilizing Co/Al(2)O(3)/Co magnetic tunnel junctions with Co electrodes of different crystalline phases, a clear relationship between electrode crystal structure and junction transport properties is presented. For junctions with one fcc(111) textured and one polycrystalline (polyphase and polydirectional) Co electrode, a strong asymmetry is observed in the magnetotransport properties, while when both electrodes are polycrystalline the magnetotransport is essentially symmetric. These observations are successfully explained within a model based on ballistic tunneling between the calculated band structures (density of states) of fcc-Co and hcp-Co. PMID- 11909385 TI - Ferroelectric domain structure of SrBi2Nb2O9 epitaxial thin films. AB - The domain structure in a ferroelectric with well-defined crystallography and negligible ferroelastic distortion (<0.002%) is reported. In contrast to prototypical ferroelectrics in which long-range elastic strain dictates the domain structure, in SrBi2Nb2O9 the elastic term is insignificant, allowing dipole-dipole interactions and domain wall energies to dominate in determining the domain structure. Electron microscopy reveals ferroelectric domains that are irregularly shaped and highly curved. Out-of-phase boundary defects are shown to be weakly correlated with 90 degrees ferroelectric domain structure. PMID- 11909386 TI - Anisotropy and magnetic field effects on the entanglement of a two qubit Heisenberg XY chain. AB - We investigate the entanglement of a two-qubit anisotropic Heisenberg XY chain in thermal equilibrium at temperature T in the presence of an external magnetic field B along the z axis. By means of the combined influences of anisotropic interactions and a magnetic field B, one is able to produce entanglement for any finite T, by adjusting the magnetic field strength. This contrasts with the isotropic interaction or the B = 0 cases, for which there is no entanglement above a critical temperature T(c) that is independent of the external B field. PMID- 11909387 TI - The stress tensor in entangled polymers. AB - In entangled polymeric liquids, a subchain connecting two entanglements is an open system which can exchange particles (Kuhn segments) with its neighboring subchains along the polymer chain. We present a calculation of the subchain mechanical behavior as determined from the grand canonical formalism of statistical mechanics, with different subchains in the same chain sharing the same chemical potential. It is shown that the linear monomer density is a constant, as originally inferred by Doi and Edwards, but the tension is generally different from one subchain to another. Accounting for the latter effect leads to a new tensorial strain measure, somewhat different from that proposed by Doi and Edwards. PMID- 11909388 TI - Fluctuation-driven dynamics of the internet topology. AB - We study the dynamics of the Internet topology based on empirical data on the level of the autonomous systems. It is found that the fluctuations occurring in the stochastic process of connecting and disconnecting edges are important features of the Internet dynamics. The network's overall growth can be described approximately by a single characteristic degree growth rate g(eff) approximately 0.016 and the fluctuation strength sigma(eff) approximately 0.14, together with the vertex growth rate alpha approximately 0.029. A stochastic model which incorporates these values and an adaptation rule newly introduced reproduces several features of the real Internet topology such as the correlations between the degrees of different vertices. PMID- 11909389 TI - Riemannian geometry of irrotational vortex acoustics. AB - We consider acoustic propagation in an irrotational vortex, using the technical machinery of differential geometry to investigate the "acoustic geometry" that is probed by the sound waves. The acoustic space-time curvature of a constant circulation hydrodynamical vortex leads to deflection of phonons at appreciable distances from the vortex core. The scattering angle for phonon rays is shown to be quadratic in the small quantity Gamma/2pi(cb), where Gamma is the vortex circulation, c the speed of sound, and b the impact parameter. PMID- 11909390 TI - Diffraction of a superfluid fermi gas by an atomic grating. AB - An atomic grating generated by a pulsed standing-wave laser field is proposed to manipulate the superfluid state in a quantum degenerate gas of fermionic atoms. We show that in the presence of atomic Cooper pairs, the density oscillations of the gas caused by the atomic grating exhibit a much longer coherence time than that in the normal Fermi gas. Our result indicates that the technique of a pulsed atomic grating is a potential candidate to detect the atomic superfluid state in a quantum degenerate Fermi gas. PMID- 11909391 TI - Spontaneous symmetry breaking with abnormal number of Nambu-Goldstone bosons and kaon condensate. AB - We describe a class of relativistic models incorporating a finite density of matter in which spontaneous breakdown of continuous symmetries leads to a lesser number of Nambu-Goldstone bosons than that required by the Goldstone theorem. This class, in particular, describes the dynamics of the kaon condensate in the color-flavor locked phase of high density QCD. We describe the spectrum of low energy excitations in this dynamics and show that, despite the presence of a condensate and gapless excitations, this system is not a superfluid. PMID- 11909392 TI - Shadows of the planck scale: scale dependence of compactification geometry. AB - By studying the effects of the shape moduli associated with toroidal compactifications, we demonstrate that Planck-sized extra dimensions can cast significant "shadows" over low-energy physics. These shadows distort our perceptions of the compactification geometry associated with large extra dimensions and place a fundamental limit on our ability to probe the geometry of compactification by measuring Kaluza-Klein states. We also find that compactification geometry is effectively renormalized as a function of energy scale, with "renormalization group equations" describing the "flow" of geometric parameters such as compactification radii and shape angles as functions of energy. PMID- 11909393 TI - Covariant coordinate transformations on noncommutative space. AB - We show how to define gauge-covariant coordinate transformations on a noncommuting space. The construction uses the Seiberg-Witten equation and generalizes similar results for commuting coordinates. PMID- 11909394 TI - Observation of the decay K- --> pi(-)mu(+)mu(-) and measurements of the branching ratios for K+/- --> pi(+/-)mu(+)mu(-). AB - Using data collected with the HyperCP (E871) spectrometer during the 1997 fixed target run at Fermilab, we report the first observation of the decay K--->pi( )mu(+)mu(-) and new measurements of the branching ratios for K+/--->pi(+/ )mu(+)mu(-). By combining the branching ratios for the decays K+-->pi(+)mu(+)mu( ) and K--->pi(-)mu(+)mu(-), we measure Gamma(K+/--->pi(+/-)mu(+)mu(-))/Gamma(K+/- ->all) = (9.8+/-1.0+/-0.5)x10(-8). The CP asymmetry between the rates of the two decay modes is [Gamma(K+-->pi(+)mu(+)mu(-))-Gamma(K--->pi(-)mu(+)mu( ))]/[Gamma(K+-->pi(+)mu(+)mu(-))+Gamma(K--->pi(-)mu(+)mu(-))] = -0.02+/-0.11+/ 0.04. PMID- 11909395 TI - Can the minimal supersymmetric standard model with light bottom squark and light gluino survive Z-peak constraints? AB - In the framework of the minimal supersymmetric model we examine the Z-peak constraints on the scenario of one light bottom squark (sbottom) ( approximately 2-5.5 GeV) and light gluino (approximately 12-16 GeV), which has been successfully used to explain the excess of bottom quark production in hadron collisions. Such a scenario is found to be severely constrained by the CERN LEP Z peak observables, especially by R(b), due to the large effect of gluino-sbottom loops. To account for the R(b) data in this scenario, the other mass eigenstate of sbottom, i.e., the heavier one, must be lighter than 125 (195) GeV at 2sigma(3sigma) level, which, however, is disfavored by CERN LEP II experiments. PMID- 11909397 TI - Predictions for Higgs and supersymmetry spectra from SO(10) Yukawa unification with mu > 0. AB - We use t, b, tau Yukawa unification to constrain supersymmetry parameter space. We find a narrow region survives for mu>0 (suggested by b-->sgamma and the anomalous magnetic moment of the muon) with A0 approximately -1.9m(16), m(10) approximately 1.4m(16), m(16) approximately 1200-3000 GeV and muM(1/2) approximately 100-500 GeV. Demanding Yukawa unification thus makes definite predictions for Higgs and sparticle masses. PMID- 11909398 TI - Collective deceleration of ultrarelativistic nuclei and creation of quark-gluon plasma. AB - We propose a unified space-time picture of baryon stopping and quark-gluon plasma creation in ultrarelativistic heavy-ion collisions. It is assumed that the highly Lorentz contracted nuclei are decelerated by the coherent color field which is formed between them after they pass through each other. This process continues until the field is neutralized by the Schwinger mechanism. Conservation of energy and momentum allow us to calculate the energy losses of the nuclear slabs and the initial energy density of the quark-gluon plasma. PMID- 11909399 TI - 129Xe-Xe molecular spin relaxation. AB - We identify the formation of bound 129Xe-Xe molecules as the primary fundamental spin-relaxation process at densities below 14 amagat. Low pressure Xe relaxation rate measurements as a function of gas composition show that Xe-Xe molecular relaxation contributes 1/T1 = 1/4.1 h to the total observed relaxation rate. The measured rate is consistent with theoretical estimates deduced from previously measured NMR chemical shifts. At atmospheric pressure the molecular relaxation is more than an order of magnitude stronger than binary relaxation. Confusion of molecular and wall relaxation mechanisms has historically caused wall relaxation rates to be overestimated. PMID- 11909400 TI - Quantum state preparation and conditional coherence. AB - It is well known that spontaneous parametric down-conversion can be used to probabilistically prepare single-photon states. We have performed an experiment in which arbitrary superpositions of zero- and one-photon states can be prepared by appropriate postselection. The optical phase, which is meaningful only for superpositions of photon number, is related to the relative phase between the zero- and one-photon states. Whereas the light from spontaneous parametric down conversion has an undefined phase, we show that this technique collapses one beam to a state of well-defined optical phase when a measurement succeeds on the other beam. PMID- 11909396 TI - Search for CP violation in tau--> K(pi)nu(tau) decays. AB - We search and find no evidence for CP violation in tau decays into the K(pi)nu(tau) final state. We provide limits on the imaginary part of the coupling constant Lambda describing a relative contribution of the CP violating processes with respect to the standard model to be -0.172 approximately l(zeta)p exhibit anomalous scaling: zeta(p) = p / 2 rather than the expected zeta(p) = p / 3. Correspondingly, the energy spectrum is described by E(k) approximately k(-2) rather than the expected E(k) approximately k(-5/3). PMID- 11909406 TI - Phonon anomalies due to collective stripe modes in high T(c) cuprates. AB - Phonon anomalies observed in various high T(c) cuprates by neutron experiments are analyzed theoretically in terms of the stripe concept. The phonon self-energy correction is evaluated by taking into account the charge collective modes of stripes, giving rise to dispersion gap, or kink and shadow phonon modes at twice the wave number of spin stripe. These features coincide precisely with observations. The gapped branches of the phonon are found to be in-phase and out of-phase oscillations relative to the charge collective mode. PMID- 11909407 TI - Nearly space-filling fractal networks of carbon nanopores. AB - Small-angle x-ray scattering, nitrogen adsorption, and scanning tunneling microscopy show that a series of activated carbons host an extended fractal network of channels with dimension D(p) = 2.8-3.0 (pore fractal), channel width 15-20 A (lower end of scaling), network diameter 3000-3400 A (upper end of scaling), and porosity of 0.3-0.6. We interpret the network as a stack of quasiplanar invasion percolation clusters, formed by oxidative removal of walls between closed voids of diameter of approximately 10 A and held in registry by fibrils of the biological precursor, and point out unique applications. PMID- 11909408 TI - Smectic membranes in motion: approaching the fast limits of x-ray photon correlation spectroscopy. AB - The dynamics of the layer-displacement fluctuations in smectic membranes have been studied by x-ray photon correlation spectroscopy (XPCS). We report transitions from an oscillatory damping regime to simple exponential decay of the fluctuations, both as a function of membrane thickness and upon changing from specular to off-specular scattering. This behavior is in agreement with recent theories. Employing avalanche photodiode detectors and the uniform filling mode of the synchrotron storage ring, the fast limits of XPCS have been explored down to 50 ns. PMID- 11909409 TI - Structural identification of metcars. AB - The ground-state structure of the metcar Ti8C12 is investigated using first principles computer simulations. Comparison with recent experimental data on the vibrational spectrum of gas phase Ti8C12 allows one to identify the geometrical structure of the clusters studied in the experiment. The present combination of predictive first-principles computer simulations and detailed experimental measurements of the vibrational spectra of clusters offers the first viable tool for structural identification of cluster shapes. PMID- 11909410 TI - Stress field in granular systems: loop forces and potential formulation. AB - The transmission of stress through a marginally stable granular pile in two dimensions is exactly formulated in terms of a vector field of loop forces, and thence in terms of a single scalar potential. This leads to a local constitutive equation coupling the stress tensor to fluctuations in the local geometry. For a disordered pile of rough grains this means the stress tensor components are coupled in a frustrated manner. In piles of rough grains with long range staggered order, frustration is avoided and a simple linear theory follows. We show that piles of smooth grains can be mapped onto a pile of unfrustrated rough grains, indicating that the problems of rough and smooth grains may be fundamentally distinct. PMID- 11909411 TI - Chemical isomerism as a key to explore free-energy landscapes in disordered matter. AB - The effects of a minor chemical modification on the microscopic structure of a material in its glass and crystal phases are investigated by the concurrent use of neutron diffraction and computer simulation. Significant changes in short-, intermediate-, and long-range order are found, resulting from the change in molecular structure. These differences are explainable by a shift in the balance between directional and excluded-volume interactions. PMID- 11909412 TI - Solution of a multiple-scattering inverse problem: electron diffraction from surfaces. AB - We present a solution to the multiple-scattering inverse problem for low-energy electron diffraction that enables the determination of the three-dimensional atomic structure of an entire surface unit cell directly from measured data. The solution requires a knowledge of the structure of the underlying bulk crystal and is implemented by a maximum entropy algorithm. PMID- 11909413 TI - Zero-point vibration of hydrogen adsorbed on Si and Pt surfaces. AB - Hydrogen atoms adsorbed on Si(111) and Pt(111) were investigated by nuclear reaction analysis (NRA) using 1H(15N,alphagamma)12C. From measurements of the NRA spectrum at normal and tilted ion incidences the zero-point vibrational energies of H on Si(111) in the perpendicular and parallel directions were found to be 123.4+/-4.6 and 44.6+/-6.2 meV, respectively, which are consistent with harmonic potentials. The zero-point energies obtained for Pt(111)-H were 80.8+/-3.9 and 62.1+/-6.0 meV for perpendicular and parallel directions, respectively. These results indicate that the stretching mode is harmonic, while the bending mode is strongly anharmonic. PMID- 11909414 TI - Mean-field nucleation theory with nonlocal interactions. AB - Mean-field nucleation theory has for a long time been successfully used to extract microscopic parameters from island density data in growth experiments. However, it produces grossly incorrect results when used to analyze weakly corrugated systems, where adsorbate interactions cannot be neglected. Here, a mean-field theory that includes nonlocal adsorbate interactions is developed and successfully tested against kinetic Monte Carlo growth simulations for a realistic adsorbate system. PMID- 11909415 TI - Novel continuum modeling of crystal surface evolution. AB - We propose a novel approach to continuum modeling of the dynamics of crystal surfaces. Our model follows the evolution of an ensemble of step configurations, which are consistent with the macroscopic surface profile. Contrary to the usual approach where the continuum limit is achieved when typical surface features consist of many steps, our continuum limit is approached when the number of step configurations of the ensemble is very large. The model can handle singular surface structures such as corners and facets. It has a clear computational advantage over discrete models. PMID- 11909416 TI - Coulomb drag shot noise in coupled Luttinger liquids. AB - Coulomb drag shot noise has been studied theoretically for 1D interacting electron systems, which are realized, e.g., in single-wall nanotubes. We show that under adiabatic coupling to external leads, the Coulomb drag shot noise of two coupled or crossed nanotubes contains surprising effects, in particular, a complete locking of the shot noise in the tubes. In contrast to Coulomb drag of the average current, the noise locking is based on a symmetry of the underlying Hamiltonian and is not limited to asymptotically small energy scales. PMID- 11909417 TI - Hall effect induced by a spin-polarized current in superconductors. AB - We propose a novel anomalous Hall effect caused by the spin-polarized current in superconductors (SC). The spin-polarized quasiparticles flowing in SC are deflected by spin-orbit scattering to yield a quasiparticle charge imbalance in the transverse direction. Overall charge neutrality gives rise to a compensating change in the number of Cooper pairs. A transverse electric field builds up as opposed to an acceleration of the Cooper pairs, producing the Hall voltage. It is found that the Hall voltages due to the side jump and skew scattering mechanisms have different temperature dependence in the superconducting state. A spin injection Hall device to generate the ac Josephson effect is proposed. PMID- 11909418 TI - Optical intersubband transitions and femtosecond dynamics in Ag/Fe(100) quantum wells. AB - The optical intersubband transitions and femtosecond dynamics of electrons in quantum well states in Ag/Fe(100) are investigated by interferometric time resolved two-photon photoemission. The quantum well wave functions and transition probabilities are evaluated from the two-photon photoemission resonance energies and intensities using an extended phase accumulation model. Direct femtosecond pump-probe correlation measurements elucidate the importance of interfaces in confined structures. PMID- 11909419 TI - Shot noise through a quantum dot in the Kondo regime. AB - The shot noise in the current through a quantum dot is calculated as a function of voltage from the high-voltage Coulomb-blockaded regime to the low-voltage Kondo regime. Using several complementary approaches, it is shown that the zero frequency shot noise (scaled by the voltage) exhibits a nonmonotonic dependence on voltage, with a peak around the Kondo temperature. Beyond giving a good estimate of the Kondo temperature, it is shown that the shot noise yields additional information on the effects of electronic correlations on the local density of states in the Kondo regime, unaccessible in traditional transport measurements. PMID- 11909420 TI - Density-induced interchange of anisotropy axes at half-filled high Landau levels. AB - We observe density-induced 90 degrees rotations of the anisotropy axes in transport measurements at half-filled high Landau levels in the two dimensional electron system, where stripe states are proposed ( nu = 9/2, 11/2, etc.). Using a field effect transistor, we find the transition density to be 2.9x10(11) cm(-2) at nu = 9/2. Hysteresis is observed in the vicinity of the transition. We construct a phase boundary in the filling factor magnetic field plane in the regime 4.4 or approximately 10(-10)M(4)(Pl) untenable. PMID- 11909444 TI - Bound states of string networks and D-branes. AB - We show the existence of nonthreshold bound states of (p,q) string networks and D3-branes, preserving 1/4 of the full type-IIB supersymmetry, interpreted as string networks "dissolved" in D3-branes. We also explicitly write down the expression for the mass density of the system and discuss the extension of the construction to other Dp-branes. Differences in our construction of string networks with the ones interpreted as dyons in N = 4 gauge theories are also pointed out. PMID- 11909445 TI - M-theory conifolds. AB - Seven manifolds of G2 holonomy provide a bridge between M-theory and string theory, via Kaluza-Klein reduction to Calabi-Yau six manifolds. We find first order equations for a new family of G2 metrics D7, with S3 x S3 principal orbits. These are related at weak string coupling to the resolved conifold, paralleling earlier examples B7 that are related to the deformed conifold, allowing a deeper study of topology change and mirror symmetry in M-theory. The D7 metrics' nontrivial parameter characterizes the squashing of an S3 bolt, which limits to S2 at weak coupling. In general the D7 metrics are asymptotically locally conical, with a nowhere-singular circle action. PMID- 11909447 TI - Solution of the multi-Reggeon compound state problem in multicolor QCD. AB - We study the properties of the color-singlet compound states of Reggeized gluons in multicolor QCD using their relation with non-compact two-dimensional Heisenberg spin magnets. Applying the methods of integrable models, we calculate their spectrum and discuss the application of the obtained results to high-energy asymptotics of the scattering amplitudes in perturbative QCD. PMID- 11909448 TI - Three-jet cross sections in hadron-hadron collisions at next-to-leading order. AB - We present a new QCD-event generator for hadron colliders which can calculate one , two-, and three-jet cross sections at next-to-leading order accuracy. We study the transverse energy spectrum of three-jet hadronic events using the k(radially) algorithm. We show that the next-to-leading order correction significantly reduces the renormalization and factorization scale dependence of the three-jet cross section. PMID- 11909449 TI - Deeply bound 1s and 2p pionic states in 205Pb and determination of the s-wave part of the pion-nucleus interaction. AB - We observed well-separated 1s and 2p pi(-) states in 205Pb in the 206Pb(d,3He) reaction at T(d) = 604.3 MeV. The binding energies and the widths determined are B(1s) = 6.762+/-0.061 MeV, Gamma(1s) = 0.764(+0.154)(-0.171) MeV, B(2p) = 5.110+/ 0.045 MeV, and Gamma(2p) = 0.321(-0.062)(+0.060) MeV. They are used to deduce the real and imaginary strengths of the s-wave part of the pion-nucleus interaction, which translates into a positive mass shift of pi(-) in 205Pb. PMID- 11909450 TI - Suppression of soft nuclear bremsstrahlung in proton-nucleus collisions. AB - Photon energy spectra up to the kinematic limit have been measured in 190 MeV proton reactions with light and heavy nuclei to investigate the influence of the multiple-scattering process on the photon production. Relative to the predictions of models based on a quasifree production mechanism, a strong suppression of bremsstrahlung is observed in the low-energy region of the photon spectrum. We attribute this effect to the interference of photon amplitudes due to multiple scattering of nucleons in the nuclear medium. PMID- 11909451 TI - Revalidation of the isobaric multiplet mass equation. AB - We have determined the energy of the J(pi) = 1/2(+), T = 3/2 resonance in 32S(p,p) to be E(p) = 3374.7+/-0.8 keV. This disagrees with the previously accepted value of E(p) = 3370+/-1 keV by Abbondanno et al. [Nuovo Cimento 70A, 391 (1970)] and solves a problem raised by recent observations of unexpected deviations from the isobaric multiplet mass equation. This resonance is also important in calibrating the beta-delayed proton spectra from 33Ar and 32Ar, and our findings may modify previous conclusions. PMID- 11909452 TI - Mechanical and chemical spinodal instabilities in finite quantum systems. AB - Self-consistent quantum approaches are used to study the instabilities of finite nuclear systems. The frequencies of multipole density fluctuations are determined as a function of dilution and temperature for several isotopes. The spinodal region of the phase diagrams is determined, and it appears that instabilities are reduced by finite size effects. The role of surface and volume instabilities is discussed. It is indicated that the important chemical effects associated with mechanical disruption may lead to isospin fractionation. PMID- 11909453 TI - Interaction induced localization in a gas of pyramidal molecules. AB - We propose a model to describe a gas of pyramidal molecules interacting via dipole-dipole interactions. The interaction modifies the tunneling properties between the classical equilibrium configurations of the single molecule and, for sufficiently high pressure, the molecules become localized in these classical configurations. We explain quantitatively, without free parameters, the shift to zero frequency of the inversion line observed upon increase of the pressure in a gas of ammonia or deuterated ammonia. For sufficiently high pressures, our model suggests the existence of a super-selection rule for states of different chirality in substituted derivatives. PMID- 11909454 TI - Optical detection of nonradiating alkali atoms in solid helium. AB - We have detected by optical means nonfluorescing 85Rb and 87Rb atoms implanted in a body centered cubic 4He crystal. In contrast to cesium the resonance fluorescence of rubidium is strongly quenched by the helium matrix, and the weak resonance absorption of the two Rb isotopes was detected using a double resonance technique. From a comparative study of the (optically detected) magnetic resonance spectra of 85Rb, 87Rb, and 133Cs we infer their effective g(F) factors and conclude that they are not perturbed by the He matrix at a level of 2 x 10( 4). We show further that optical pumping of Rb proceeds via depopulation, whereas for Cs it proceeds via repopulation. PMID- 11909455 TI - Effect of relativistic many-electron interactions on photoelectron partial wave probabilities. AB - We obtain relative cross sections for the production of photoelectrons with specific angular momentum quantum numbers. These cross sections are obtained from the polarization analysis of the visible fluorescence of ions produced when circularly polarized vacuum ultraviolet radiation photoionizes ground state Ar. The ratio of cross sections for the production of photoelectrons with the same orbital angular momentum but different total angular momenta shows strong deviations from the statistical ratio, demonstrating the importance of relativistic interactions in many-electron photoionization dynamics. PMID- 11909446 TI - Q2 Dependence of quadrupole strength in the gamma*p --> Delta(+)(1232) --> p pi(0) transition. AB - Models of baryon structure predict a small quadrupole deformation of the nucleon due to residual tensor forces between quarks or distortions from the pion cloud. Sensitivity to quark versus pion degrees of freedom occurs through the Q2 dependence of the magnetic (M1+), electric (E1+), and scalar (S1+) multipoles in the gamma*p-->Delta(+)-->p pi(0) transition. We report new experimental values for the ratios E(1+)/M(1+) and S(1+)/M(1+) over the range Q2 = 0.4-1.8 GeV2, extracted from precision p(e,e(')p)pi(0) data using a truncated multipole expansion. Results are best described by recent unitary models in which the pion cloud plays a dominant role. PMID- 11909457 TI - Reflection-type hologram for atoms. AB - A cold metastable neon atomic beam was manipulated with a reflective amplitude hologram that was encoded on a silicon surface. A black-and-white pattern of atoms was reconstructed on a microchannel plate detector. The hologram used the enhanced quantum reflection developed by authors and was made of a two dimensional array of rectangular low and high reflective cells. The surface of the high reflective cell was composed of regularly spaced roof-shaped ridges, while the low reflective cell was simply a flat surface. The hologram was the first demonstration of reflective atom-optical elements that used universal interaction between a neutral atom and solid surface. PMID- 11909456 TI - Coherent transient enhancement of optically induced resonant transitions. AB - By applying pulse shaping techniques to a broadband 100 fs pulse in resonance with a two-level atomic transition, we are able to enhance the peak transient excited level population relative to that achievable with transform limited pulses. We also demonstrate how the dispersion induced by the absorption line itself leads to similar rapidly oscillating transients in the excited population. These transient population effects are applicable in any multiphoton resonant transition. PMID- 11909458 TI - Efficient generation of flexible-monomer intermolecular potential energy surfaces. AB - A new method of generating flexible-monomer intermolecular interaction potentials has been proposed. The method, based on symmetry-adapted perturbation theory, extends a rigid-monomer potential into a flexible-monomer one at a cost negligible compared to performing calculations on a full-dimensional grid (i.e., including internal degrees of freedom of monomers). The non-rigidity effects are accounted for by density-overlap integrals and by asymptotic expansion coefficients. Results for a model system (Ar-HF) demonstrate that the method recovers a substantial portion of these effects. PMID- 11909459 TI - HF dimer in small helium clusters: interchange-tunneling dynamics in a quantum environment. AB - We present diffusion quantum Monte Carlo calculations of the interchange tunneling splitting of (4)He(n)(HF)(2) clusters, n = 1-10. The tunneling splitting decreases rapidly for n = 1-4 clusters, and much more slowly for n>4. The decrease calculated for (4)He(n)(HF)(2) represents 74% of the reduction in the tunneling splitting measured recently for HF dimer in nanodroplets of more than 2000 He atoms. The first four He atoms quench the interchange tunneling very efficiently by virtue of occupying the equatorial ring which encircles the C(2h) transition state of the tunneling pathway. PMID- 11909460 TI - Optical trapping and manipulation of nano-objects with an apertureless probe. AB - We propose a novel way to trap and manipulate nano-objects above a dielectric substrate using an apertureless near-field probe. A combination of evanescent illumination and light scattering at the probe apex is used to shape the optical field into a localized, three-dimensional optical trap. We use the coupled-dipole method and the Maxwell stress tensor to provide a self-consistent description of the optical force, including retardation and the influence of the substrate. We show that small objects can be selectively captured and manipulated under realistic conditions. PMID- 11909461 TI - Spatial-field correlation: the building block of mesoscopic fluctuations. AB - The absence of self-averaging in mesoscopic systems is a consequence of long range intensity correlations. Microwave measurements suggest, and diagrammatic calculations confirm, that the correlation function of the normalized intensity with displacement of the source and detector, Delta R and Delta r, respectively, can be expressed as the sum of three terms, with distinctive spatial dependences. Each term involves only the sum or the product of the square of the field correlation function, F identical with F(2)(E). The leading-order term is the product, F(Delta R)F(Delta r); the next term is proportional to the sum, F(Delta R)+F(Delta r); the third term is proportional to F(Delta R)F(Delta r)+[F(Delta R)+F(Delta r)]+1. PMID- 11909462 TI - Degree of polarization in near fields of thermal sources: effects of surface waves. AB - We introduce the concept of degree of polarization for electromagnetic near fields. The approach is based on the generalized Stokes parameters that appear as expansion coefficients of the 3 x 3 coherence matrix in terms of the Gell-Mann matrices. The formalism is applied to optical near fields of thermally fluctuating half-space sources with particular interest in fields that are strongly polarized owing to resonant surface plasmons or phonons. This novel method is particularly useful when assessing the full vectorial characteristics of random evanescent fields, e.g., for near-field spectroscopy and polarization microscopy. PMID- 11909463 TI - Transition to chaos in continuous-time random dynamical systems. AB - We consider situations where, in a continuous-time dynamical system, a nonchaotic attractor coexists with a nonattracting chaotic saddle, as in a periodic window. Under the influence of noise, chaos can arise. We investigate the fundamental dynamical mechanism responsible for the transition and obtain a general scaling law for the largest Lyapunov exponent. A striking finding is that the topology of the flow is fundamentally disturbed after the onset of noisy chaos, and we point out that such a disturbance is due to changes in the number of unstable eigendirections along a continuous trajectory under the influence of noise. PMID- 11909464 TI - Variational approach to hard sphere segregation under gravity. AB - It is demonstrated that the minimization of the free energy functional for hard spheres and hard disks yields the result that excited granular materials under gravity segregate not only in the widely known "Brazil nut" fashion, i.e., with the larger particles rising to the top, but also in reverse "Brazil nut" fashion. Specifically, the local density approximation is used to investigate the crossover between the two types of segregation occurring in the liquid state, and the results are found to agree qualitatively with previously published results of simulation and of a simple model based on condensation. PMID- 11909465 TI - Single-shot electron-beam bunch length measurements. AB - We report subpicosecond electro-optic measurements of the length of individual relativistic electron bunches. The longitudinal electron-bunch shape is encoded electro-optically on to the spectrum of a chirped laser pulse. The electron-bunch length is determined by analyzing individual laser-pulse spectra obtained with and without the presence of an electron bunch. Since the length of the chirped laser pulse can be easily changed, the electron bunch can be visualized on different time scales. This single-shot imaging technique is a promising method for real-time electron-bunch diagnostics. PMID- 11909466 TI - Hosing and sloshing of short-pulse GeV-class wakefield drivers. AB - This Letter examines the electron-hosing instability in relation to the drivers of current and future plasma-wakefield experiments using fully three-dimensional particle-in-cell simulation models. The simulation results are compared to numerical solutions and to asymptotic solutions of the idealized analytic equations. The measured growth rates do not agree with the existing theory and the behavior is shown to depend sensitively on beam length, shape, and charge. We find that even when severe hosing occurs the wake can remain relatively stable. PMID- 11909467 TI - Competition between two forms of ordering in finite Coulomb clusters. AB - The lowest-energy state of spherical clusters made up of single-species charged particles in a three-dimensional confining potential is investigated by molecular dynamics simulations for a system size of 5 x 10(3) to 1.2 x 10(5). The energy per particle is compared between shell-structured clusters and spherical finite bcc lattices with relaxed surfaces. The shell structure in the interior is the lowest-energy configuration for ion numbers lower than about 10(4), while for higher ion numbers, an interior with bcc ordering surrounded by a few shells on the outside has lower energy. The formation of a small bcc lattice (nucleation) in the shell-structured cluster of 2 x 10(4) ions is observed. PMID- 11909468 TI - Laser-generated waves and wakes in rotating ion crystals. AB - Locally excited plasma waves are generated in a Coulomb crystal by "pushing" with radiation pressure on a rotating cloud of laser-cooled 9Be+ ions. The waves form a stationary wake that is directly imaged through the dependence of the ion fluorescence on Doppler shifts, and theoretical calculations in a slab geometry are shown to accurately reproduce these images. The technique demonstrates a new method of exciting and studying waves in cold ion clouds. PMID- 11909469 TI - Suppression of MHD fluctuations leading to improved confinement in a gun-driven spheromak. AB - Magnetic fluctuations have been reduced to approximately 1% during discharges on the Sustained Spheromak Physics Experiment by shaping the spatial distribution of the bias magnetic flux in the device. In the resulting quiescent regime, the safety factor profile is nearly flat in the plasma and the dominant ideal and resistive MHD modes are greatly reduced. During this period, the temperature profile is peaked at the magnetic axis and maps onto magnetic flux contours. Energy confinement time is improved over previous reports in a driven spheromak. PMID- 11909470 TI - Neoclassical tearing physics in the spherical tokamak MAST. AB - Results from MAST provide a first test of neoclassical tearing mode physics in the spherical tokamak (ST). The mode accounts for the main performance limit in conventional tokamaks. Its behavior in the ST is remarkably well described by existing theoretical models, although it is more readily seeded by sawtooth events in these scenarios. Modeling confirms the significance of stabilizing field-curvature effects. This provides good grounds for optimism that with suitable control of profiles, it may be possible to avoid these modes in the ST. PMID- 11909471 TI - Topology of electronic charge density and energetics of planar faults in fcc metals. AB - Using ab initio calculations we have studied the energetics and the evolution of the electronic charge density with shear in three fcc metals exhibiting different deformation properties, aluminum, silver, and iridium. The charge redistribution described by the change in character of specific charge density critical points (cps), is ascertained from the values of the charge density, rho(0), and its three principal curvatures, rho( parallel parallel), rho(hh), and rho(vv), respectively. The change in character of cps correlates with the energetics. For all three metals, rho(hh) vanishes near the unstable stacking configuration. The symmetry or asymmetry of the charge redistribution, measured by rho(hh)/rho(vv), may be an important factor determining stacking fault energies. PMID- 11909472 TI - Dynamics of sodium in sodium disilicate: channel relaxation and sodium diffusion. AB - We use molecular-dynamics computer simulations to study the dynamics of amorphous (Na2O)2(SiO2). We find that the Na ions move in channels embedded in a SiO2 matrix. The characteristic distance between these channels gives rise to a prepeak in the structure factor at q approximately equal to 0.95 A(-1). The dynamics of sodium is given by a fast process which can be seen in the incoherent scattering function and a slow process which is seen in the coherent function. The relaxation time of the latter coincides with the alpha-relaxation time of the matrix. The Kohlrausch exponent of the fast process for q>1.6 A(-1) is the same as the von Schweidler exponent for the slow one. Thus the two processes are closely related. PMID- 11909473 TI - Multiscale modeling of precipitate microstructure evolution. AB - We demonstrate how three "state-of-the-art" techniques may be combined to build a bridge between atomistics and microstructure: (1) first-principles calculations, (2) a mixed-space cluster expansion approach, and (3) the diffuse-interface phase field model. The first two methods are used to construct the driving forces for a phase-field microstructural model of theta'- Al2Cu precipitates in Al: bulk, interfacial, and elastic energies. This multiscale approach allows one to isolate the physical effects responsible for precipitate microstructure evolution. PMID- 11909474 TI - Novel pathway to the growth of diamond on cubic beta-SiC(001). AB - By carrying out first-principles calculations on diamond-forming processes, we predict a method for the heteroepitaxial growth of diamond on cubic beta SiC(001). In the method, we used two processes: (i) the preformation of an sp(3) like surface configuration of beta-SiC(001) by the adsorption of group-V surfactants; (ii) the successive growth of diamond by the segregation of the surfactants onto a surface and the desorption of surface hydrogen. Analyzing the segregation energies, we found that the atomic size effect plays a crucial role in the surfactant-mediated growth of diamond on beta-SiC(001). PMID- 11909475 TI - Computer simulation of displacement cascades in nanocrystalline ni. AB - Large-scale molecular dynamics of cascade production of the primary damage state are performed in fcc nanocrystalline Ni of average grain diameters of 5 and 12 nm. Primary knock-on atom kinetic energies of 5-30 keV are simulated. During the thermal spike phase, significant atomic motion towards the surrounding grain boundary structure is observed, characterized by many replacement-collision sequences. Upon resolidification, the excess volume condenses to form vacancy dominated defects with a complex partial dislocation network forming at higher energies. PMID- 11909476 TI - Hydrogen vibration modes in GaP:N: the pivotal role of nitrogen in stabilizing the H(2)(*) complex. AB - Atomic structures of N-related hydrogen complexes in GaP:N are calculated from first principles. As the more electronegative N bonds H stronger than P, it stabilizes the H(2)(*) complex that is otherwise unstable against the formation of an H2 molecule. This provides the first theoretical proof that H(2)(*) can be stable in a III-V semiconductor. The previously proposed H-N-H dihydride model is found to be unstable against spontaneously transforming into H(2)(*), which involves only monohydrides, H-N and H-Ga. The calculated local vibrational frequencies and isotope shifts are in good agreement with experiment. PMID- 11909478 TI - Surface modification without desorption: recycling of Cl on Si(100)-(2 x 1). AB - We demonstrate chlorine-induced modification of Si(100)-(2 x 1) under conditions where Cl is recycled rather than desorbed as SiCl2. A dimer with 2 Cl atoms, 2SiCl, converts to SiCl2+Si, allowing the bare Si atom to escape onto the terrace. At temperatures below the desorption threshold, the SiCl2 unit decays through Cl diffusion, allowing the second Si atom to escape. The result is a dimer vacancy, terrace regrowth structures, and Cl that is able to participate in another pitting event. Access to this unexpected roughening pathway is controlled by the Cl concentration and temperature. This previously overlooked process represents an important component of Si(100) surface processing. PMID- 11909477 TI - Outstanding magnetic properties of nematic suspensions of goethite (alpha-FeOOH) nanorods. AB - Aqueous suspensions of goethite (alpha-FeOOH) nanorods form a mineral lyotropic nematic phase that aligns in a very low magnetic field (20 mT for samples 20 microm thick). The particles orient along the field direction at intensities smaller than 350 mT, but they reorient perpendicular to the field beyond 350 mT. This outstanding behavior is also observed in the isotropic phase which has a very strong magnetic-field induced birefringence that could be interesting for applications. We interpret these magnetic effects as resulting from a competition between a nanorod remanent magnetic moment and a negative anisotropy of its magnetic susceptibility. PMID- 11909479 TI - Marginal scaling scenario and analytic results for a glassy compaction model. AB - A diffusion-deposition model for glassy dynamics in compacting granular systems is treated by time scaling and by a method that provides the exact asymptotic (long-time) behavior. The results include Vogel-Fulcher dependence of rates on density, inverse logarithmic time decay of densities, exponential distribution of decay times, and broadening of noise spectrum. These are all in broad agreement with experiments. The main characteristics result from a marginal rescaling in time of the control parameter (density); this is argued to be generic for glassy systems. PMID- 11909480 TI - Oxygen diffusion through the disordered oxide network during silicon oxidation. AB - An atomic-scale description is provided for the long-range oxygen migration through the disordered SiO2 oxide during silicon oxidation. First-principles calculations, classical molecular dynamics, and Monte Carlo simulations are used in sequence to span the relevant length and time scales. The O2 molecule is firmly identified as the transported oxygen species and is found to percolate through interstices without exchanging oxygen atoms with the network. The interstitial network for O2 diffusion is statistically described in terms of its potential energy landscape and connectivity. The associated activation energy is found in agreement with experimental values. PMID- 11909481 TI - Cation mass dependence of the nearly constant dielectric loss in alkali triborate glasses. AB - Electrical ac conductivity measurements on alkali triborate glasses ( M2O x 3B2O3, M = Li, Na, K, and Rb) were performed at temperatures down to 8 K and frequencies up to 1 GHz. All samples show a nearly constant dielectric loss (NCL), at the limit of high frequencies and/or low temperatures. The magnitude of the NCL is found to decrease as m(-1/3) with increasing alkali ion mass m. This quantitative result for the NCL, closely related to the mean-square displacement of ions, indicates that the origin of the NCL might be related to vibrational relaxation of the ions in the anharmonic potentials that cage them, and the cage is decaying very slowly with time. PMID- 11909482 TI - Overlayer strain relief on surfaces with square symmetry: phase diagram for a 2D Frenkel-Kontorova model. AB - Overlayers on surfaces with square symmetry exhibit a huge variety of strain relief mechanisms. I present a simple 2D Frenkel-Kontorova model and calculate the associated zero temperature phase diagram which shows a transition from overlayers with square symmetry (and possible square dislocation patterns) to hexagonal symmetry. The phase diagram includes the experimentally observed clock rotated phase. Local density approximation calculations suggested by the model show that a clean Ni(100) surface reconstructs from a bulk-terminated to a clock rotated structure at biaxial compressive strains above 2.5%. PMID- 11909483 TI - Freezing of a stripe liquid. AB - The existence of a stripe-liquid phase in a layered nickelate, La(1.725)Sr(0.275)NiO(4), is demonstrated through neutron scattering measurements. We show that incommensurate magnetic fluctuations evolve continuously through the charge-ordering temperature, although an abrupt decrease in the effective damping energy is observed on cooling through the transition. The energy and momentum dependence of the magnetic scattering are parametrized with a damped-harmonic-oscillator model describing overdamped spin waves in the antiferromagnetic domains defined instantaneously by charge stripes. PMID- 11909484 TI - Direct observation of charge order in an epitaxial NdNiO3 film. AB - The first direct observation of charge order of Ni(3+delta(')) and Ni(3-delta) by resonant x-ray scattering experiments in an epitaxial film of NdNiO3 is reported. A quantitative value of delta+delta(') = (0.45 +/- 0.04)e was obtained. The temperature dependence of the charge order deviates significantly from those of the magnetic moment and crystallographic structure. This might be an indication of a difference in their fluctuation time scales. These observations are discussed in terms of the temperature-driven metal-insulator transition in the RNiO3 family. PMID- 11909485 TI - First-order pairing transition and single-particle spectral function in the attractive hubbard model. AB - A dynamical mean-field theory analysis of the attractive Hubbard model in the normal phase is carried out upon restricting to solutions where superconducting order is not allowed. A clear first-order pairing transition as a function of the coupling takes place at all the electron densities out of half filling between a Fermi liquid, stable for UU(c), and it is accompanied by phase separation. The spectral function in the metallic phase is constituted by a low-energy structure around the Fermi level, which disappears discontinuously at U = U(c), and two high-energy features (Hubbard bands), which persist in the insulating phase. PMID- 11909486 TI - Theory of ferromagnetism in Ca(1-x)La(x)B(6). AB - Novel ferromagnetism in Ca(1-x)La(x)B(6) is studied in terms of the Ginzburg Landau theory for excitonic-order parameters, taking into account symmetry of the wave functions. We found that the minima of the free energy break both inversion and time-reversal symmetries, while the product of these two remains preserved. This explains various novelties of the ferromagnetism and predicts a number of magnetic properties, including the magnetoelectric effect, which can be tested experimentally. PMID- 11909487 TI - Spin-filter device based on the Rashba effect using a nonmagnetic resonant tunneling diode. AB - We propose an electronic spin-filter device that uses a nonmagnetic triple barrier resonant tunneling diode (TB-RTD). This device combines the spin-split resonant tunneling levels induced by the Rashba spin-orbit interaction and the spin blockade phenomena between two regions separated by the middle barrier in the TB-RTD. Detailed calculations using the InAlAs/InGaAs material system reveal that a splitting of a peak should be observed in the I-V curve of this device as a result of the spin-filtering effect. The filtering efficiency exceeds 99.9% at the peak positions in the I-V curve. PMID- 11909488 TI - Shell filling and exchange coupling in metallic single-walled carbon nanotubes. AB - We report the characterization of electronic shell filling in metallic single walled carbon nanotubes by low-temperature transport measurements. Nanotube quantum dots with average conductance approximately (1-2)e(2)/h exhibit a distinct four-electron periodicity for electron addition as well as signatures of Kondo and inelastic cotunneling. The Hartree-Fock parameters that govern the electronic structure of metallic nanotubes are determined from the analysis of transport data using a shell-filling model that incorporates the nanotube band structure and Coulomb and exchange interactions. PMID- 11909489 TI - Dominance of Fermi-surface holes in p-type tunneling. AB - In-plane uniaxial stress is used to tune continuously the mixing between the heavy-hole (HH) and light-hole (LH) states in a p-type double-barrier structure. The LH1 and HH2 resonant tunneling peaks shift at almost the same rate with stress, in contrast to the corresponding exciton peaks observed by photoreflectance, which exhibit a strong Fano-related anticrossing. Comparison between the observed shifts and a four-band k x p calculation of the state energies in the well provides the first experimental proof that the flow of holes through off-zone center states dominates the resonant tunneling current in p-type structures. PMID- 11909490 TI - Two-stage Kondo effect in a quantum dot at a high magnetic field. AB - We report a strong Kondo effect (Kondo temperature approximately 4 K) at high magnetic field in a selective area growth semiconductor quantum dot. The Kondo effect is ascribed to a singlet-triplet transition in the ground state of the dot. At the transition, the low-temperature conductance approaches the unitary limit. Away from the transition, for low bias voltages and temperatures, the conductance is sharply reduced. The observed behavior is compared to predictions for a two-stage Kondo effect in quantum dots coupled to single-channel leads. PMID- 11909491 TI - Observation of quantized Hall drag in a strongly correlated bilayer electron system. AB - The frictional drag between parallel two-dimensional electron systems has been measured in a regime of strong interlayer correlations. When the bilayer system enters the excitonic quantized Hall state at total Landau level filling factor nu(T) = 1, the longitudinal component of the drag vanishes but a strong Hall component develops. The Hall drag resistance is observed to be accurately quantized at h/e(2). PMID- 11909492 TI - Electronic transport through carbon nanotubes: effects of structural deformation and tube chirality. AB - Atomistic simulations using a combination of classical force field and density functional theory (DFT) show that carbon atoms remain essentially sp(2) coordinated in either bent tubes or tubes pushed by an atomically sharp atomic force microscope (AFM) tip. Subsequent Green's-function-based transport calculations reveal that for armchair tubes there is no significant drop in conductance, while for zigzag tubes the conductance can drop by several orders of magnitude in AFM-pushed tubes. The effect can be attributed to simple stretching of the tube under tip deformation, which opens up an energy gap at the Fermi surface. PMID- 11909493 TI - Prediction of high T(c) superconductivity in hole-doped LiBC. AB - The layered lithium borocarbide LiBC, isovalent with and structurally similar to the superconductor MgB2, is an insulator due to the modulation within the hexagonal layers (BC vs B2). We show that hole doping of LiBC results in Fermi surfaces of B-C p sigma character that couple very strongly to B-C bond stretching modes, precisely the features that lead to superconductivity at T(c) approximately equal to 40 K in MgB2. Comparison of Li(0.5)BC with MgB2 indicates the former to be a prime candidate for electron-phonon coupled superconductivity at substantially higher temperature than in MgB2. PMID- 11909494 TI - Evidence for two-band superconductivity from break-junction tunneling on MgB2. AB - Superconductor-insulator-superconductor tunnel junctions have been fabricated on MgB2 that display Josephson and quasiparticle currents. These junctions exhibit a gap magnitude, Delta approximately 2.5 meV, that is considerably smaller than the BCS value, but which clearly and reproducibly closes near the bulk T(c). In conjunction with fits of the conductance spectra, these results are interpreted as direct evidence of two-band superconductivity. PMID- 11909495 TI - Exact correlation functions of the BCS model in the canonical ensemble. AB - We evaluate correlation functions of the BCS model for a finite number of particles. The integrability of the Hamiltonian relates it with the Gaudin algebra G[sl(2)]. Therefore, a theorem that Sklyanin proved for the Gaudin model, can be applied. Several diagonal and off-diagonal correlators are calculated. The finite-size scaling behavior of the pairing correlation function is studied. PMID- 11909497 TI - Coherent magnetic oscillation in the spin ladder system alpha(')- NaV2O5. AB - We report the first observation of coherent magnetic excitations in a spin ladder system NaV2O5 by using femtosecond time-domain spectroscopy. A pronounced coherent oscillation is observed at 127 cm(-1) (nearly twice the spin gap energy) and assigned to a two-magnon bound state, based on the temperature dependence of the intensity below the charge ordering phase transition at T(C) = 34 K. This mode can be observable only when circularly polarized light is used as a pump or a probe beam, suggesting that it corresponds to a spin-flip excitation from the singlet ground state. A phonon mode strongly coupled to the spin state is also found at 303 cm(-1). PMID- 11909496 TI - Quantal phases, disorder effects, and superconductivity in spin-Peierls systems. AB - In view of recent developments in the investigation on cuprate high- T(c) superconductors and the spin-Peierls compound CuGeO3, we study the effect of dilute impurity doping on the spin-Peierls state in quasi-one-dimensional systems. We identify a common origin for the emergence of antiferromagnetic order upon the introduction of static vacancies and superconductivity for mobile holes. PMID- 11909498 TI - Gate-induced band ferromagnetism in an organic polymer. AB - We propose that a chain of five-membered rings (polyaminotriazole) should be ferromagnetic with an appropriate doping that is envisaged to be feasible with a field-effect transistor structure. The ferromagnetism is confirmed by a spin density functional calculation, which also shows that ferromagnetism survives the Peierls instability. We explain the magnetism in terms of the Mielke and Tasaki flatband ferromagnetism with the Hubbard model. This opens a new possibility of band ferromagnetism in purely organic polymers. PMID- 11909499 TI - Evolution of magnetic polarons and spin-carrier interactions through the metal insulator transition in Eu(1-x)Gd(x)O. AB - Raman scattering studies as functions of temperature, magnetic field, and Gd substitution are used to investigate the evolution of magnetic polarons and spin carrier interactions through the metal-insulator transition in Eu(1-x)Gd(x)O. These studies reveal a spin-fluctuation-dominated paramagnetic (PM) regime for T>T*>T(C), and a coexistence regime for T9% of patients. The incidence and levels of hypertension associated with COX 2 inhibitors are within the range of those observed with nonspecific NSAIDs. Apparent differences between the COX-2 inhibitors celecoxib and rofecoxib may be functions of differences in study population susceptibilities to NSAID-mediated hypertensive effects. Patients at risk for hypertension should be monitored for changes in blood pressure during NSAID treatment. PMID- 11909558 TI - Cardiovascular pharmacology of nonselective nonsteroidal anti-inflammatory drugs and coxibs: clinical considerations. AB - Cyclooxygenase (COX)-2 inhibitors have been developed with the goal of providing similar efficacy and greater safety compared with traditional nonsteroidal anti inflammatory drugs. Development was based on the hypothesis that COX-1 is the housekeeping enzyme necessary for production of prostaglandins (PGs) with homeostatic functions, whereas COX-2 is a mediator of pathophysiologic processes. However, later research has demonstrated a role of COX-2 in production of PGs that have functions under normal physiologic conditions. In the vasculature, COX 2 seems to be the main enzyme responsible for the production of prostacyclin. Increased synthesis of this vasodilatory and antithrombotic PG represents a homeostatic response during periods of accelerated platelet-vessel wall interactions and counteracts increased synthesis of COX-1-derived prothrombotic prostanoid thromboxane A(2) (TXA(2)). The clinical sequelae of inhibiting prostacyclin activity in the absence of concomitant inhibition of TXA(2) are not currently clear. Animal studies show that inhibition of prostacyclin activity does not lead to spontaneous thrombosis but may increase response to thrombotic stimuli. Therefore, prostacyclin synthesis may be important for limiting thrombotic events in patients who are at an increased cardiovascular risk. Overviews of clinical studies in arthritis and Alzheimer's disease have not demonstrated increased cardiovascular risk associated with specific COX-2 inhibition in most patients. However, data from 1 clinical trial revealed a 5 fold divergence in rates of myocardial infarction between a coxib and a nonsteroidal anti-inflammatory drug comparitor. Credible explanations for the results of this trial have been proposed and further studies are necessary to clarify the relative risk-to-benefit ratio of COX-2 inhibition in patients at increased risk for cardiovascular events, and the effects of concomitant aspirin therapy. PMID- 11909559 TI - Relative risk of cardiovascular events in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is associated with increased comorbidity and mortality resulting from cardiovascular disease. A review of past and recent studies suggests that inflammation and thrombosis may provide a link between both diseases. This association has significant clinical implications for therapy, because many of the drugs used in the symptomatic treatment of RA, such as nonsteroidal anti-inflammatory drugs and the new cyclooxygenase (COX)-2-specific inhibitors, affect mediators of both inflammation and thrombosis. Recent studies have elucidated the potential effects of COX-2-specific inhibitors on thrombogenic events and their potential impact on cardiovascular disease. Although a causal effect of these drugs to increase the risk of cardiovascular disease has not been established, further investigation is needed on the effects of COX-2-specific inhibitors on cardiovascular risk. Such data would be of particular importance in determining appropriate therapeutic approaches to the treatment of patients with RA who are also at risk for cardiovascular events. PMID- 11909560 TI - Extent of, and factors associated with, delay to hospital presentation in patients with acute coronary disease (the GRACE registry). AB - Our primary study aim was to examine extent of, and factors associated with, delay in seeking medical care in a large multinational registry of patients with acute myocardial infarction (AMI) and unstable angina pectoris. A secondary goal was to examine the relation between duration of prehospital delay and receipt and timing of coronary reperfusion strategies. Investigators from 14 countries are participating in the Global Registry of Acute Coronary Events (GRACE) project. The study sample consisted of 3,693 patients with ST-segment elevation AMI, 2,935 with non-ST-segment elevation AMI, and 3,954 patients with unstable angina hospitalized between 1999 and 2001. The average and median delay times were longest in patients with non-ST-segment elevation AMI (6.1 and 3.0 hours, respectively) followed by patients with unstable angina (5.6 and 3.0 hours) and those with ST-segment elevation AMI (4.7 and 2.3 hours). Approximately 41% of patients with ST-segment elevation AMI presented to the 94 study hospitals within 2 hours of the onset of acute coronary symptoms; this compared with approximately one third of patients with non-ST-segment elevation AMI and unstable angina. Several demographic and clinical characteristics were associated with prehospital delay. In patients with ST-segment elevation AMI, duration of prehospital delay was inversely related to the receipt of thrombolytic therapy, but was inconsistently related to the use of percutaneous coronary interventions. The results of this study demonstrate that a large proportion of patients continue to exhibit prolonged delay in seeking medical care after the onset of acute coronary symptoms and remain in need of targeted educational efforts to reduce extent of delay. PMID- 11909561 TI - Comparison of results of coronary angioplasty for acute myocardial infarction in patients > or =75 years of age versus patients <75 years of age. AB - We reviewed 1,063 consecutive patients treated with direct coronary angioplasty for acute myocardial infarction (AMI): 261 were > or =75 and 802 were <75 years of age. Compared with the younger group, the older group had a higher percentage of women (48% vs 22%, p <0.0001), multivessel coronary disease (50% vs 39%, p <0.01), overall in-hospital mortality (8.4% vs 3.7%, p <0.01), cardiac mortality rate (6.1% vs 3.1%, p <0.05), and noncardiac mortality rate (2.3% vs 0.6%, p <0.05). Successful reperfusion was achieved in both groups at a similarly high rate (93% and 95%, p = NS). Hospital mortality was similar whether reperfusion was successful or failed. Successful compared with unsuccessful angioplasty decreased mortality rates in the older (6.6% vs 33%, p <0.0001) and younger (3.0% vs 18%, p <0.0001) groups. When reperfusion was successful, the cardiac mortality rate in older patients was not significantly higher than in younger patients: 4.1% vs 2.4%, p = NS. PMID- 11909562 TI - Effects of gold coating of coronary stents on neointimal proliferation following stent implantation. AB - Experimental studies suggest a reduced neointimal tissue proliferation in vascular stainless steel stents coated with gold. This prospective multicenter trial evaluated the impact of gold coating on neointimal tissue proliferation in patients undergoing elective stent implantation. The primary end point was the in stent tissue proliferation measured by intravascular ultrasound at 6 months comparing stents of identical design with or without gold coating (Inflow). Two hundred four patients were randomized to receive uncoated (group A, n = 101) or coated (group B, n = 103) stents. Baseline parameters did not differ between the groups. Stent length and balloon size were comparable, whereas inflation pressure was slightly higher in group A (14 +/- 3 vs 13 +/- 3 atm, p = 0.013). Procedural success was similar (A, 97%; B, 96%). The acute angiographic result was better for group B (remaining stenosis 4 +/- 12% vs 10 +/- 11%, p = 0.002). Six-month examinations revealed more neointimal proliferation in group B. By ultrasound, the neointimal volume within the stent was 47 +/- 25 versus 41 +/- 23 mm(3) (p = 0.04), with a ratio of neointimal volume-to-stent volume of 0.45 +/- 0.12 versus 0.40 +/- 0.12 (p = 0.003). The angiographic minimal luminal diameter was smaller in group B (1.47 +/- 0.57 vs 1.69 +/- 0.70 mm, p = 0.04), with a higher late luminal loss of 1.17 +/- 0.51 versus 0.82 +/- 0.56 mm (p = 0.001). Thus, gold coating of the tested stent type resulted in more neointimal tissue proliferation. PMID- 11909563 TI - Haptoglobin phenotype and the risk of restenosis after coronary artery stent implantation. AB - We recently demonstrated that an allelic polymorphism in the haptoglobin gene is a major determinant of susceptibility to a number of vascular disorders. We set out to determine if haptoglobin phenotype was predictive of the development of restenosis in a consecutive series of patients, all of whom underwent stent implantation followed by repeat angiography with quantitative coronary angiography analysis 6 months later. This study included 214 consecutive patients undergoing stent implantation for de novo lesions between 1998 and 1999 in Aalst, Belgium. All underwent follow-up quantitative coronary angiography analysis 6 months after the procedure. The haptoglobin phenotype was determined by electrophoresis. No significant differences were found between patients segregated by phenotype with respect to clinical, procedural, and angiographic factors previously suggested to influence the development of restenosis. None of the diabetic patients homozygous for the haptoglobin 1 allele developed restenosis compared with a >50% restenosis rate for diabetic patients with at least 1 haptoglobin 2 allele (p <0.02). In all patients (diabetic and nondiabetic), we observed a trend toward a lower incidence of restenosis in patients homozygous for the 1 allele (21% vs 33%, p <0.09). Moreover, we found a graded risk relation to the number of haptoglobin 2 alleles. The risk of developing restenosis was greater in subjects with 2 haptoglobin 2 alleles (36%) than in those with 1 haptoglobin 2 allele (31%) or no haptoglobin 2 alleles (21%). Thus, knowledge of the haptoglobin phenotype may be useful in assessing and utilizing new therapies that attempt to reduce restenosis, and may have important implications for the risk stratification algorithm used in managing diabetic patients with coronary artery disease. PMID- 11909564 TI - Usefulness of rest and low-dose dobutamine wall motion scores in predicting survival and benefit from revascularization in patients with ischemic cardiomyopathy. AB - This study examined the value of wall motion scores at rest and with low- and high-dose dobutamine infusion for prediction of outcome and benefit from revascularization in patients with ischemic cardiomyopathy. Follow-up was obtained in 139 patients with ischemic cardiomyopathy who had echocardiography at rest, and during low- (10 microg/kg/min) and high-dose dobutamine (maximal dose 50 microg/kg/min) infusion. Both rest and low-dose wall motion scores were multivariate predictors of cardiac death, but ischemia and peak dose scores were not predictors. Rest scores risk stratified patients into 3 groups: score (1.00 to 1.99) with 11% cardiac death; score (2.00 to 2.49) with 30% death; and score > or =2.50 with 47% death. One third of patients with rest scores > or =2.50 had improvement in scores to < 2.50 with low-dose dobutamine. Their frequency of cardiac death was reduced to 23% compared with 60% (p = 0.04) in those who remained with low-dose scores > or =2.50. Low-dose scores also identified those who benefited from revascularization. In patients with low-dose scores (1.00 to 1.99), the frequency of cardiac death was marginally lower in revascularized than nonrevascularized patients (10% vs 21%, p = 0.28). In patients with scores (2.00 to 2.49), revascularized patients had a significantly lower frequency of cardiac death than nonrevascularized patients (15% vs 41%, p < 0.05). The frequency of death in those with low-dose scores > or =2.50 was very high in both revascularized (75%) and nonrevascularized (56%, p = 0.42) patients. Thus, rest and low-dose wall motion scores enable risk stratification of patients with ischemic cardiomyopathy and identify those who do and do not benefit from revascularization. PMID- 11909566 TI - Effects of enhanced external counterpulsation on stress radionuclide coronary perfusion and exercise capacity in chronic stable angina pectoris. AB - Enhanced external counterpulsation (EECP) is an effective noninvasive treatment for patients with coronary artery disease (CAD). EECP has been demonstrated to improve anginal class and time to ST-segment depression during exercise stress testing. This study assesses the efficacy of EECP in improving stress-induced myocardial ischemia using radionuclide perfusion treadmill stress tests (RPSTs). The international study group enrolled patients from 7 centers with chronic stable angina pectoris and a baseline ischemic pre-EECP RPST. Patients' demographic and clinical characteristics were recorded. A baseline pre-EECP maximal RPST was performed within 1 month before EECP treatment. The results were compared with a follow-up RPST performed within 6 months of completion of a 35 hour course of EECP. Four centers performed post-EECP RPST to the same level of exercise as pre-EECP, whereas 3 centers performed maximal RPST post-EECP. The study enrolled 175 patients (155 men and 20 women). Improvement in angina, defined by > or =1 Canadian Cardiovascular Society angina class change, was reported in 85% of patients. In the centers performing the same level of exercise, 81 of 97 patients (83%) had significant improvement in RPST perfusion images. Patients who underwent maximal RPST revealed improvement in exercise duration (6.61 +/- 1.88 pre-EECP vs 7.41 +/- 2.03 minutes post-EECP, p <0.0001); 42 of the 78 patients (54%) in this group showed significant improvement in RPST perfusion images. Thus, EECP was effective in improving stress myocardial perfusion in patients with chronic stable angina at both comparable (baseline) and at maximal exercise levels. PMID- 11909565 TI - Usefulness of dobutamine Tc-99m sestamibi-gated single-photon emission computed tomography for prediction of left ventricular ejection fraction outcome after coronary revascularization for ischemic cardiomyopathy. AB - Gated single-photon emission computed tomography (SPECT) imaging allows analysis of myocardial perfusion and assessment of baseline global and regional left ventricular (LV) function and their changes during low-dose dobutamine infusion. The study examined whether the changes in LV ejection fraction induced by dobutamine and evaluated using technetium-99m sestamibi- gated SPECT predict the evolution of ejection fraction after revascularization in patients with ischemic cardiomyopathy. Thirty-seven patients underwent resting and dobutamine nitrate enhanced sestamibi-gated SPECT before revascularization and baseline-resting sestamibi gated SPECT after intervention to assess global functional changes. A postrevascularization improvement in ejection fraction > or =5 U was defined as significant. At follow-up, ejection fraction increased significantly in 19 patients. According to receiver-operating characteristic curve analysis, an increase in ejection fraction > or =5 U during dobutamine was the optimal cutoff value for predicting a significant postrevascularization improvement, with 79% sensitivity, 78% specificity, and 78% accuracy. A significant correlation was found between dobutamine and postrevascularization ejection fraction (r = 0.85; p <0.0001). The increase in ejection fraction during dobutamine is a good predictor of an improvement in ejection fraction after revascularization. This represents another important diagnostic contribution obtained using gated SPECT imaging for the assessment of myocardial viability in patients with ischemic cardiomyopathy. PMID- 11909567 TI - Relation of lipoprotein(a) as coronary risk factor to type 2 diabetes mellitus and low-density lipoprotein cholesterol in patients > or =65 years of age (The Italian Longitudinal Study on Aging). AB - High levels of serum lipoprotein(a) [Lp(a)] have been associated with increased risk of coronary artery disease (CAD), but this association apparently is not confirmed in elderly people. We evaluated the interactions of Lp(a) with lipid and nonlipid CAD risk factors in a sample of subjects enrolled in the prevalence survey (1992 to 1993) of the Italian Longitudinal Study on Aging (ILSA). The entire population consisted of 5,632 elderly people, aged 65 to 84 years, randomly selected in 8 Italian municipalities. The present cross-sectional study included 400 free-living elderly subjects (74 +/- 6 years) from the randomized cohort of Casamassima (Bari, Southern Italy) (n = 704). The results showed that in the elderly population, high serum Lp(a) is a CAD risk factor dependent on type 2 diabetes mellitus and elevated low-density lipoprotein (LDL) cholesterol levels. In particular, the combined effect of high Lp(a) (> or =20 mg/dl) and high LDL cholesterol (> or =3.63 mmol/L [> or =140 mg/dl]), increases coronary risk by 2.75 (95% confidence interval 7.70 to 0.99); finally, the effect of Lp(a) > or =20 mg/dl and LDL cholesterol > or =3.63 mmol/L (> or =140 mg/dl), combined with type 2 diabetes mellitus, increases risk of CAD by 6.65 (95% confidence interval 35.40 to 1.25). In the elderly, elevated Lp(a) levels appear not to be an independent predictor of CAD, but this lipoprotein is a risk factor only in subjects with type 2 diabetes mellitus and elevated LDL cholesterol. PMID- 11909568 TI - Changes in hemodynamics and left ventricular structure after menopause. AB - To evaluate the cardiovascular changes associated with menopause, we studied hemodynamics at rest, ambulatory blood pressure, and left ventricular structure in a biracial cohort of pre- and postmenopausal women of similar age, race, weight, and blood pressure. Despite similar levels of blood pressure, postmenopausal women had a higher indexed peripheral resistance (2,722 +/- 757 vs 2,262 +/- 661 dynes.s.m(2)/cm(5), p <0.01) and a lower cardiac index (2.64 +/- 0.73 vs 3.10 +/- 0.71 L/min.m(2), p <0.01) than premenopausal women. Postmenopausal women also had less nocturnal decreases in both systolic (15 +/- 8 vs 19 +/- 8 mm Hg, p <0.01) and diastolic (12 +/- 6 vs 15 +/- 6 mm Hg, p = 0.05) pressures during ambulatory monitoring and higher levels of hematocrit (40 +/- 2% vs 38 +/- 3%, p <0.01). In association with this greater hemodynamic load, postmenopausal women had evidence of early concentric left ventricular remodeling, manifested by a greater relative wall thickness (0.38 +/- 0.06 vs 0.35 +/- 0.06, p <0.01) than that observed in premenopausal women. Differences between pre- and postmenopausal women in hemodynamics, diurnal blood pressure variation, and left ventricular structure were observed in white and African American subjects. These results suggest that menopause is associated with hemodynamic changes and left ventricular remodeling, which may contribute to the enhanced cardiovascular risk observed in postmenopausal women. PMID- 11909569 TI - Usefulness of plasma vitamin B(6), B(12), folate, homocysteine, and creatinine in predicting outcomes in heart transplant recipients. AB - Atherothrombotic complications are frequently seen in patients undergoing heart transplantation. These patients have high plasma total homocysteine concentrations associated with lower folate and vitamin B(6) levels. The relation between these metabolic abnormalities and the development of vascular complications, however, remains unclear. Fasting plasma total homocysteine, folate, vitamin B(12), vitamin B(6), and creatinine were measured in 160 cardiac transplant recipients who were followed for a mean duration of 28 +/- 9 months after blood draw (mean 59 +/- 28 months after transplant). Cardiovascular events and causes of mortality were determined and Cox proportional-hazards regression analysis was used to identify the independent predictors for cardiovascular events and mortality. Twenty-five patients developed cardiovascular events and 17 died (11 cardiovascular deaths). Mean +/- SD total homocysteine value was 18.4 +/ 8.5 (range 4.3 to 63.5 micromol/L). Hyperhomocysteinemia (> or =15 micromol/L) was seen in 99 patients (62%). Levels were no different in patients with or without cardiovascular complications/death (16.8 +/- 6.2 vs 18.9 +/- 9 micromol/L, p = 0.4). However, vitamin B(6) deficiency was seen in 21% of recipients with and in 9% without cardiovascular complications/death (p = 0.05). The relative risk for cardiovascular events, including cardiovascular death, increased 2.7 times (confidence interval 1.2 to 5.9) for B(6) levels < or =20 nmol/L compared with those with normal B(6) levels (p = 0.02). Thus, hyperhomocysteinemia is common in transplant recipients but may have no causal role in the atherothrombotic vascular complications of transplantation. Deficiency of vitamin B(6), however, may predict adverse outcomes, suggesting a possible role for supplementation with this vitamin. PMID- 11909570 TI - Prognostic value of electrocardiograms, ventricular late potentials, ventricular arrhythmias, and left ventricular systolic dysfunction in patients with Duchenne muscular dystrophy. AB - Myocardial involvement is a common finding in patients with Duchenne-type muscular dystrophy (DMD). Nevertheless, the prognostic values of standard electrocardiogram (ECG), ventricular arrhythmias, ventricular late potentials (LPs), and left ventricular (LV) systolic dysfunction have not been extensively investigated. Eighty-four patients with DMD (aged 18.6 +/- 4.8 years) underwent standard and signal-averaged electrocardiography, 24-hour Holter monitoring, and echocardiography. The prevalence of electrocardiographic abnormalities, frequent ventricular premature complexes, LPs, and LV systolic dysfunction was 71%, 32%, 28%, and 35%, respectively. Median follow-up was 76 months (range 5 to 106). The mortality rate in the follow-up period was 27%. The typical DMD electrocardiographic alterations, ventricular arrhythmic pattern, and LPs were not predictors of mortality. In contrast, the presence of LV systolic dysfunction detected on echocardiography was a powerful predictor of mortality in the follow up period (p = 0.013, hazard ratio 3.14, 95% confidence interval 1.27 to 7.79). Thus, echocardiographic assessment of LV systolic dysfunction provides prognostic information in patients with DMD. Electrocardiographic alterations, ventricular arrhythmias, and LPs have no prognostic value in predicting mortality in these patients. PMID- 11909571 TI - Management of hypoplastic left heart syndrome in the 1990s. AB - We used the discharge database of the University Hospital Consortium to determine the management and early outcome of neonates with hypoplastic left heart syndrome admitted to member institutions from 1990 to 1999. Of the 2,264 patients, 1,203 underwent a Norwood procedure, with 42% mortality. Cardiac transplantation was performed in 72, with 38% mortality, and 217 (10%) were discharged without any surgical procedure. The proportion of patients managed by the Norwood procedure increased from 43% during the first half of the decade to 59% in the second half, with corresponding decreases in the proportion managed by transplantation or nonintervention. A mortality rate of < or =40% was achieved in all 5 institutions performing >50 Norwood procedures, and by 9 of 40 institutions performing <50. Performance of a Norwood procedure has become the most frequent management for neonates with hypoplastic left heart syndrome. Lower operative mortality rates are generally, but not exclusively, achieved by institutions with high surgical volume. PMID- 11909572 TI - Validation of an algorithm for predicting cardiac events in renal transplant candidates. AB - A 2-tiered noninvasive cardiac risk stratification algorithm was first evaluated in a test population with planar thallium myocardial perfusion imaging and subsequently in a validation population using single-photon emission computed tomographic (SPECT) thallium myocardial perfusion imaging. This study examines if SPECT imaging was as predictive of cardiac death as planar imaging and also if SPECT imaging predicted nonfatal cardiac events in the patient population. Renal transplant candidates were evaluated using a 2-tiered noninvasive cardiac risk stratification algorithm. The first tier of risk assessment utilized 5 variables: age >50 years, insulin-dependent diabetes mellitus, abnormal electrocardiogram, and a history of either angina or congestive heart failure. Patients without risk factors were considered low risk and underwent no further cardiac evaluation. Patients with > or =1 risk factor were considered high risk and underwent a second tier of risk assessment with planar (n = 95) or SPECT (n = 112) imaging. In the test population, 13 of 16 cardiac deaths (81%) occurred in high-risk patients with abnormal planar studies. In the validation group, all cardiac deaths (5 of 60) and nonfatal cardiac events (13 of 60) occurred in high-risk patients with abnormal SPECT studies. SPECT imaging was at least as predictive as planar imaging and also predicted nonfatal as well as fatal cardiac events. Pretransplant risk stratification by clinical variables identified low-risk patients who may not require further cardiac evaluation and high-risk patients with normal SPECT imaging who may not require angiography. PMID- 11909573 TI - Usefulness of pulse deficit to predict in-hospital complications and mortality in patients with acute type A aortic dissection. AB - Vascular compromise seen with pulse deficits is common in patients with type A dissection. However, patient characteristics and in-hospital outcomes associated with pulse deficits have not been evaluated. Accordingly, we studied 513 patients (mean age 62 +/- 14 years, 65% men) with acute type A aortic dissection enrolled in the International Registry of Acute Aortic Dissection. Pulse deficits, defined as decreased or absent carotid or peripheral pulses as noted by clinicians and later confirmed by diagnostic imaging, at surgery or at autopsy were noted in 154 patients (30%). Age <70 years, male gender, neurologic deficit(s), altered mental status, and hypotension, shock, or tamponade on admission were all significantly higher in patients with than without pulse deficits. The etiology of aortic dissection, clinical symptoms, and imaging findings were similar in the 2 groups. In-hospital complications (hypotension, coma, renal failure, and limb ischemia) and mortality (41% vs 25%, p = 0.0002) were significantly higher in patients with pulse deficit. Cox proportional-hazards regression analysis identified pulse deficit as an independent predictor of 5-day in-hospital mortality (risk ratio 2.73, 95% confidence interval 1.7 to 4.4; p <0.0001). Further, overall mortality rates increased with an increasing number of pulse deficits (p for trend <0.0001). Pulse deficits are common findings in patients with type A aortic dissection and identify those at high risk of in-hospital adverse events. This simple clinical sign should direct physicians to consider a diagnosis of aortic dissection in patients with acute chest pain, and should help identify a subgroup of patients who would benefit from more aggressive strategies. PMID- 11909574 TI - What if it were your child? PMID- 11909575 TI - Comparison of low-density lipoprotein cholesterol lowering by pravastatin to <100 mg/dl versus >100 mg/dl on brachial artery vasoreactivity in patients with severe hypercholesterolemia and previous atherosclerotic events or diabetes mellitus. PMID- 11909576 TI - Design of the Pravastatin or Atorvastatin Evaluation and Infection Therapy (PROVE IT)-TIMI 22 trial. PMID- 11909577 TI - Effect of oral administration of testosterone on brachial arterial vasoreactivity in men with coronary artery disease. PMID- 11909578 TI - Variations in collagen content of asynergic left ventricular segments in explanted hearts of men with ischemic cardiomyopathy. PMID- 11909579 TI - Intimal hyperplasia regression from 6 to 12 months after stenting. PMID- 11909581 TI - Efficacy and tolerability of automatic nighttime atrial fibrillation shocks in patients with permanent internal atrial defibrillators. PMID- 11909580 TI - Comparison of gold-coated NIR stents with uncoated NIR stents in patients with coronary artery disease. PMID- 11909582 TI - Accuracy of clinical diagnosis of left heart valvular or obstructive lesions in pediatric outpatients with heart murmur. PMID- 11909583 TI - Usefulness of decreased levels of D-dimer in predicting progressive accelerated graft arteriosclerosis. PMID- 11909584 TI - Effect of intensive resistance training on isotonic exercise Doppler indexes of left ventricular systolic function. PMID- 11909585 TI - Safety of subxyphoid pericardial access using a blunt-tip needle. PMID- 11909586 TI - Relation between brachial artery reactivity and noninvasive large and small arterial compliance in healthy volunteers. PMID- 11909587 TI - Fatal pulmonary toxicity occurring within two weeks of initiation of amiodarone. PMID- 11909588 TI - Two husbands--one disease: the broad clinical spectrum of hypertrophic cardiomyopathy. PMID- 11909589 TI - Synthesis of GD3-lactam: a potential ligand for the development of an anti melanoma vaccine. AB - The novel sialyl donor methyl (ethyl 4,7,8,9-tetra-O-acetyl-5-N,N-diacetylamino 3,5-dideoxy-2-thio-3-thiophenyl-D-erythro-beta-L-gluco-non-2-ulopyranosid)onate was used for glycosylation of a lactosyl acceptor to give the GM3-trisaccharide derivative in 83% yield. Introduction of an azido group at C-9" of the GM3 trisaccharide derivative, transformation into a glycosyl acceptor, and sialylation with the above mentioned novel sialyl donor gave a GD3-trisaccharide in 50% yield. Reduction of the azido group gave the corresponding amine, which underwent spontaneous lactamization to the GD3-[1"'-9"]-lactam in an overall yield of 86%. Removal of protecting groups of over five steps, followed by per-O acetylation gave an acetylated GD3-[1"'-9"]-lactam TMSEt glycoside in 27% overall yield. The acetylated GD3-[1"'-9"]-lactam TMSEt glycoside is suitable for glycosylation of linker-arms and the resulting linker-glycosides are planned to be coupled to carrier proteins, thus providing immunogens for trial vaccinations against malignant melanoma. PMID- 11909590 TI - A substrate-unspecified glycosylation reaction promoted by copper(II) trifluoromethanesulfonate in benzotrifluoride. AB - A glycosylation reaction induced by copper(II) trifluoromethanesulfonate is described. Using benzotrifluoride as the reaction solvent, five kinds of glycosyl donors, a glucosyl chloride, a fluoride, a trichloroacetimidate, a 1-O-acetyl compound, and a lactol were activated to give the corresponding glucosides. PMID- 11909591 TI - Molecular ordering of cellulose after extraction of polysaccharides from primary cell walls of Arabidopsis thaliana: a solid-state CP/MAS (13)C NMR study. AB - Solid-state CP/MAS 13C NMR spectroscopy was used to determine the effects of three different sequential extraction procedures, used to remove non-cellulosic polysaccharides, on the molecular ordering of cellulose in a cell-wall preparation containing mostly primary cell walls obtained from the leaves of the model dicotyledon, Arabidopsis thaliana. The extractions were 50 mM trans-1,2 diaminocyclohexane N,N,N',N'-tetraacetic acid (CDTA) and 50 mM sodium carbonate (giving Residue 1); 50 mM CDTA, 50 mM sodium carbonate and 1 M KOH (giving Residue 2); and 50 mM CDTA, 50 mM sodium carbonate and 4 M KOH (giving Residue 3). The molecular ordering of cellulose in Residue 1 was similar to that in unextracted walls: the cellulose was almost all crystalline, with 43% of molecules contained in crystallite interiors and similar proportions of the triclinic (I(alpha)) and monoclinic (I(beta)) crystal forms. Residue 2 was partly decrystallized and the remaining crystallites were mostly in the I(beta) form. Residue 3 was a mixture of cellulose II, cellulose I and amorphous cellulose. The presence of signals at 100.0 and 102.3 ppm in the spectra of Residues 1 and 2, but not of unextracted cell walls, suggested that the extractions giving these residues caused some of the non-cellulosic polysaccharides, possibly xyloglucans and galactoglucomannans, to become relatively well ordered, for example through interactions with cellulose crystallite surfaces. PMID- 11909592 TI - Disentangling alpha from beta mechanical relaxations in the rubber-to-glass transition of high-sugar-chitosan mixtures. AB - The occurrence of molecular motions in addition to those of the glass-transition region (alpha mechanism) were investigated in chitosan and a branched derivative substituted with alkyl chains having eight carbon atoms. Once hydrophobic interactions of the alkyl groups in aqueous solution were demonstrated, polymers were mixed with glucose syrup at high levels of solids. The real (G') and imaginary (G") components of the complex dynamic modulus in high-solid mixtures were measured between 0.1 and 100 rad s(-1) in the temperature range from -55 to 50 degrees C. The method of reduced variables gave superposed curves of G' and G", which unveiled an anomaly in the dispersion of the alkylated derivative both in terms of higher modulus values and dominant elastic component of the polymeric network, as compared with the glass-transition region of chitosan. It was proposed that the new mechanical feature was due to beta mechanism, and master curves of viscoelastic functions and relaxation processes were constructed to rationalize it. PMID- 11909593 TI - Synthesis of D-mannitol and L-iditol derivatives as monomers for the preparation of new regioregular AABB-type polyamides. AB - 1,6-Diamino-1,6-dideoxy-2,3,4,5-tetra-O-methyl-D-mannitol (and its L-iditol analogue) suitable for their utilization as monomers in the preparation of linear polyamides are described. Regio- and stereoregular polyamides of the AABB-type have been prepared by the active ester polycondensation method from these C(2) symmetric monomers and suberic and dodecanedioic acids. The resulting polyamides were obtained in fair yields (70-60%) and were characterized by elemental analyses and infrared and 1H and 13C NMR spectroscopies. Their M(w) and M(w)/M(n) were determined by GPC relative to polystyrene standards. All of them were gummy non-crystalline solids. PMID- 11909594 TI - Direct regioselective 2-O-(p-toluenesulfonylation) of sucrose. AB - 2-O-(p-Toluenesulfonyl)sucrose was regioselectively synthesized by direct p toluenesulfonylation of sucrose using N-(p-toluenesulfonyl)imidazole in the presence of molecular sieves at 40 degrees C. The reactivities of the sucrose hydroxy groups toward this sulfonylation increased in the order as follows: OH 2>>OH-1'>OH-3'>OH-6>OH-6'. These results were diametrically opposite to the expected sulfonylation with p-toluenesulfonyl chloride in pyridine, for which the reactivity increased in the order as follows: OH-6', OH-6>>OH-1'>OH-2. The desired 2-O-(p-toluenesulfonyl)sucrose was readily isolated by simple open reversed-phase column chromatography, followed by recrystallization, thus overcoming the main difficulties associated with regioselectivity, efficiency, and isolation techniques for the practical preparation. PMID- 11909595 TI - Synthesis of a potential tetrasaccharide ligand for E-selectin. AB - A potential tetrasaccharide ligand for E-selectin, (Na(+-)O(3)SO-3)Galbeta-(1- >4)[Fucalpha-(1-->3)]Glcbeta-(1-->6)Gal, an analogue of the ovarian cystadenoma glycoprotein tetrasaccharide fragment, was synthesized in a highly practical way. PMID- 11909596 TI - Glycon specificity profiling of alpha-glucosidases using monodeoxy and mono-O methyl derivatives of p-nitrophenyl alpha-D-glucopyranoside. AB - Hydrolysis of probe substrates, eight possible monodeoxy and mono-O-methyl analogs of p-nitrophenyl alpha-D-glucopyranoside (pNP alpha-D-Glc), modified at the C-2, C-3, C-4, and C-6 positions, was studied as part of investigations into the glycon specificities of seven alpha-glucosidases (EC 3.2.1.20) isolated from Saccharomyces cerevisiae, Bacillus stearothermophilus, honeybee (two enzymes), sugar beet, flint corn, and Aspergillus niger. The glucosidases from sugar beet, flint corn, and A. niger were found to hydrolyze the 2-deoxy analogs with substantially higher activities than against pNP alpha-D-Glc. Moreover, the flint corn and A. niger enzymes showed hydrolyzing activities, although low, for the 3 deoxy analog. The other four alpha-glucosidases did not exhibit any activities for either the 2- or the 3-deoxy analogs. None of the seven enzymes exhibited any activities toward the 4-deoxy, 6-deoxy, or any of the methoxy analogs. The hydrolysis results, with the deoxy substrate analogs, demonstrated that alpha glucosidases having remarkably different glycon specificities exist in nature. Further insight into the hydrolysis of deoxyglycosides was obtained by determining the kinetic parameters (k(cat) and K(m)) for the reactions of sugar beet, flint corn, and A. niger enzymes. PMID- 11909597 TI - 1-O-Acetyl-beta-D-galactopyranose: a novel substrate for the transglycosylation reaction catalyzed by the beta-galactosidase from Penicillium sp. AB - 1-O-Acetyl-beta-D-galactopyranose (AcGal), a new substrate for beta galactosidase, was synthesized in a stereoselective manner by the trichloroacetimidate procedure. Kinetic parameters (K(M) and k(cat)) for the hydrolysis of 1-O-acetyl-beta-D-galactopyranose catalyzed by the beta-D galactosidase from Penicillium sp. were compared with similar characteristics for a number of natural and synthetic substrates. The value for k(cat) in the hydrolysis of AcGal was three orders of magnitude greater than for other known substrates. The beta-galactosidase hydrolyzes AcGal with retention of anomeric configuration. The transglycosylation activity of the beta-D-galactosidase in the reaction of AcGal and methyl beta-D-galactopyranoside (1) as substrates was investigated by 1H NMR spectroscopy and HPLC techniques. The transglycosylation product using AcGal as a substrate was beta-D-galactopyranosyl-(1-->6)-1-O-acetyl beta-D-galactopyranose (with a yield of approximately 70%). In the case of 1 as a substrate, the main transglycosylation product was methyl beta-D-galactopyranosyl (1-->6)-beta-D-galactopyranoside. Methyl beta-D-galactopyranosyl-(1-->3)-beta-D galactopyranoside was found to be minor product in the latter reaction. PMID- 11909598 TI - Structure of the core part of the lipopolysaccharides from Proteus penneri strains 7, 8, 14, 15, and 21. AB - The core-lipid A region of the lipopolysaccharides from Proteus penneri strains 7, 8, 14, 15, and 21 was studied using NMR spectroscopy, ESI MS, and chemical analysis after alkaline deacylation, deamination, and mild-acid hydrolysis of the lipopolysaccharides. The following general structure of the major core oligosaccharides is proposed: [abstract: see text] where all sugars are in the pyranose form and have the D configuration unless otherwise stated, Hep and DDHep=L-glycero- and D-glycero-D-manno-heptose, respectively, K=H, and Q=H in strain 8 or alpha-Glc in strains 7, 14, 15, and 21. In addition, several minor structural variants are present, including those lacking Ara4N in strains 7 and 15 and having the alpha-GlcN residue N-acylated to a various degree with glycine in strains 7, 8, 14, and 21. In strain 14, there are also core oligosaccharides with K=amide of beta-D-GalpA with putrescine, spermidine, or 4-azaheptane-1,7 diamine; remarkably, these structural variants lack either the PEtN group or the alpha-Hep-(1-->2)-alpha-DDHep disaccharide fragment at alpha-D-GalpA. While structural features of the inner core part are shared by Proteus strains studied earlier, the outermost Q-(1-->4)-alpha-GalNAc-(1-->2)-alpha-DDHep-(1-->6)-alpha GlcN oligosaccharide unit has not been hitherto reported. PMID- 11909599 TI - Chiral induction based on carbohydrate ligands in olefin platinum(0) complexes. AB - A chiral N,N-ligand based on glucose is able to recognise selectively one enantioface of a prochiral olefin in a trigonal Pt environment. NMR and X-ray studies have been carried out aiming to disclose the factors, which govern this unexpected result. The selectivity originates from the ability of the ligand to create a chiral pocket of C(2) symmetry, which is retained in both solution and solid state. PMID- 11909600 TI - Preparation of (1-->4)-beta-D-xylooligosaccharides from an acid hydrolysate of cotton-seed xylan: suitability of cotton-seed xylan as a starting material for the preparation of (1-->4)-beta-D-xylooligosaccharides. AB - Cotton-seed residual cake, which is a byproduct of the process of oil extraction from the seed, was delignified with sodium hypochlorite (1% available chlorine). Xylan was then prepared from the delignified wet material by alkali extraction with 15% sodium hydroxide. The cotton-seed xylan contained 64.7% xylose and 9.4% uronic acid. The xylan was hydrolyzed with 0.125 M sulfuric acid at 90 degrees C for 15 min. The resultant hydrolysis products were separated by gel-permeation chromatography on BioGel P-4 and Toyopearl HW-40F columns connected in series, with water as an eluate. Xylose and xylooligosaccharides with a degree of polymerization ranging from DP 2 to 15 were separated under such conditions, and each xylooligosaccharide-containing peak fraction afforded a single band on fluorophore-assisted carbohydrate electrophoresis. These results suggest that cotton-seed xylan is suitable for the preparation of xylose and xylooligosaccharides. PMID- 11909602 TI - Pharmacogenetics in affective disorders. AB - Pharmacogenetics will be of substantial help in the field of affective disorders pharmacotherapy. The possible definition of a genetic liability profile for drug side-effects and efficacy will be of great help in treatments that need weeks to months to be effective. During the last few years, a number of groups have reported possible liability genes. The efficacy and time of onset of selective serotonin reuptake inhibitors have been associated with a polymorphism in the promoter region of the transporter (SERTPR) in many independent studies, while variants at the tryptophan hydroxylase gene, 5-HT2a receptor and G-protein beta3 have been associated with them in pilot studies. Lithium long-term prophylactic efficacy has been associated with SERTPR, TPH and inositol polyphosphate 1 phosphatase variants, though in unreplicated samples. A number of further candidate genes were not associated with these treatments. In conclusion, both acute and long-term treatments appear to be, at least to some extent, under genetic influence and preliminary data have identified possible liability genes. PMID- 11909603 TI - T-226296: a novel, orally active and selective melanin-concentrating hormone receptor antagonist. AB - Through the screening of our in-house chemical compound library, we found a novel melanin-concentrating hormone (MCH) receptor antagonist, T-226296, a (-) enantiomer of N-[6-(dimethylamino)-methyl]-5,6,7,8-tetrahydro-2-naphthalenyl]-4' fluoro[1,1'-biphenyl]-4-carboxamide. T-226296 exhibited high affinity for cloned human and rat MCH receptors (SLC-1) in receptor binding assays (IC50=5.5+/-0.12 nM for human SLC-1; 8.6+/-0.32 nM for rat SLC-1). T-226296 had high selectivity over other receptors, including the second subtype of the MCH receptor, SLT (MCH2), transporters and ion channels. In Chinese hamster ovary (CHO) cells expressing human SLC-1, T-226296 reversed the MCH-mediated inhibition of forskolin-stimulated cAMP accumulation, inhibited MCH-induced intracellular Ca2+ increase, and also inhibited MCH-stimulated arachidonic acid release. In rats, oral administration of T-226296 (30 mg/kg) almost completely suppressed the food intake induced by intracerebroventricular injection of MCH. These results clearly indicate that T-226296 is a novel, orally active and selective MCH receptor antagonist that will be promising for further exploring the physiology and pathophysiology of MCH-SLC-1 signaling. PMID- 11909604 TI - Interaction of human organic anion transporters with various cephalosporin antibiotics. AB - Cephalosporin antibiotics are thought to be excreted into the urine via organic anion transporters (OATs). The purpose of this study was to elucidate the interaction of human-OATs with various cephalosporin antibiotics, using proximal tubule cells stably expressing human-OAT1, human-OAT3 and human-OAT4. Human-OAT1 and human-OAT3 are localized to the basolateral side of the proximal tubule, whereas human-OAT4 is localized to the apical side. The cephalosporin antibiotics tested were cephalothin, cefoperazone, cefazolin, ceftriaxone, cephaloridine, cefotaxime, cefadroxil and cefamandole. All of these cephalosporin antibiotics significantly inhibited organic anion uptake mediated by human-OAT1, human-OAT3 and human-OAT4. Kinetic analysis revealed that these inhibitions were competitive. The inhibition constant (K(i)) values of cefoperazone, cefazolin, ceftriaxone and cephaloridine for human-OAT1 were much lower than those for human OAT3 and human-OAT4, whereas the K(i) values of cephalothin and cefotaxime for human-OAT3 were much lower than those for human-OAT1 and human-OAT4. Human-OAT4 mediated the bidirectional transport of estrone sulfate, an optimal substrate for human-OAT4. These results suggest that human-OAT1, human-OAT3 and human-OAT4 interact with various cephalosporin antibiotics, and that human-OAT1 and human OAT3 play a distinct role in the basolateral uptake of cephalosporin antibiotics. Since the K(i) value of cephaloridine for human-OAT4-mediated organic uptake was much higher than that for human-OAT1, the results indicate the possibility that human-OAT4 limits the efflux of cephaloridine, leading to the accumulation of cephaloridine and the induction of nephrotoxicity. PMID- 11909605 TI - Involvement of p38 mitogen-activated protein kinase in PLL-AGE-induced cyclooxgenase-2 expression. AB - In the present study, murine RAW 264.7 macrophages were incubated with poly-L lysine-derived advanced glycosylation end products (PLL-AGEs) to examine cyclooxygenase-2 protein expression. Treatment of RAW 264.7 cells with PLL-AGEs caused the dose-dependent expression of cylooxygenase-2 but not cylooxygenase-1 and an increase in cylooxygenase activity. Increased cylooxygenase-2 expression was seen at 6 h and reached a maximum at 24 h. The tyrosine kinase inhibitor, genistein, and the p38 mitogen-activated protein kinase (MAPK) inhibitor, [4-(4 fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole] (SB 203580), inhibited PLL-AGE-induced cylooxygenase-2 expression, while the Ras inhibitor, FPT inhibitor II, and the MAP kinase kinase inhibitor, (2'-amino-3' methoxyflavone) (PD 98059), had no effect on PLL-AGE-induced cylooxygenase-2 expression. Incubation of RAW 264.7 cells with PLL-AGEs resulted in activation of p38 MAPK, and this activation was suppressed by genistein and SB 203580. Taken together, our results suggest that activation of protein tyrosine kinase and p38 MAPK is involved in AGE-induced cyclooxygenase-2 expression in RAW 264.7 macrophages. PMID- 11909607 TI - Human urotensin II-induced aorta ring contractions are mediated by protein kinase C, tyrosine kinases and Rho-kinase: inhibition by somatostatin receptor antagonists. AB - Human urotensin II-(1-11) and its N-terminally shortened analogues, human urotensin II-(4-11)-OH and human urotensin II-(4-11)-NH2 are potent vasoconstrictor peptides in isolated rat thoracic aorta. Human urotensin II induced tonic aorta ring contractions are inhibited by the Ca2+ channel antagonists, verapamil, nitrendipine and diltiazem; D609 (Tricyclodecan-9-yl xanthogenate, K), selective inhibitor of phosphatidylcholine-specific phospholipase C and partially by phospholipase C inhibitor U-73122 [1-[6-((17ss-3 Methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-25-dione] and a selective inhibitor of phosphatidyl-inositol-specific phospholipase C-ET-18-OCH3 (Edelfosine,1-O-octadecyl-2O-methyl-rac-glycero-3-phosphorylcholine); protein kinase C inhibitors, chelerythrine and NPC-15437 [S-2,6-diamino-N-[[1-(1 oxotridecyl)-2-piperidinyl]methyl]-hexanamide dihydrochloride]; tyrosine kinase inhibitors, genistein and tyrphostin B42 and Rho-kinase inhibitor HA-1077 [1-(5 isoquinolinylsulfonyl)-homopiperazine dihydrochloride]. This indicates that human urotensin II-induced tonic contractions of the rat aorta are mediated by phospholipase C, protein kinase C, tyrosine kinases and Rho-kinase related pathways. In the high K+ medium, human urotensin II induces dose-dependent phasic oscillations of aortic rings. These are inhibited by Ca2+ channel antagonists, the phospholipase C inhibitor, U-73122 and protein kinase C inhibitors, chelerythrine and NPC-15437, indicating that human urotensin II-induced phasic oscillations of the rat aorta are mediated by phospholipase C and protein kinase C-dependent pathways. Given their close structural similarity, several somatostatin analogues, importantly containing DCys5 and DTrp7 and expressing different degrees of somatostatin receptor antagonist activity, were tested for possible inhibitory effects on human urotensin II-induced contractions of the rat aorta rings. Pre-incubation of rat aorta rings in the presence of somatostatin analogues, which are preferentially sst2 specific binders: PRL-2882; PRL-2903 and PRL-2915 at micro-molar concentrations significantly blocked the development of human urotensin II-induced tonic contractions. Somatostatin receptor antagonists dose-dependently inhibited human urotensin II-induced Ca2+ transients in rat thoracic aorta rings. These somatostatin receptor antagonists displayed moderate affinities for recombinant rat and human urotensin II receptor binding sites. The data support the suggestion that urotensin II receptor and somatostatin type 2/5 receptors display similar surface topologies and that analogues of somatostatin could provide useful lead compounds for the development of more potent urotensin II receptor antagonists. PMID- 11909606 TI - Long-term effects of benidipine on cerebral vasoreactivity in hypertensive rats. AB - We tested the hypothesis that long-term application of a Ca2+ channel blocker would ameliorate the functional and morphological deterioration of the cerebral arteries during hypertension. Male spontaneously hypertensive rats (SHR) were fed a standard rat chow, containing a low (3 mg/kg/day) or high dose (6 mg/kg/day) of benidipine, a Ca2+ channel blocker, for 2 months. Using a cranial window, we examined responses of the basilar artery to acetylcholine, sodium nitroprusside, (-)-(3S,4R)-4-(N-acetyl-N-hydroxyamino)-6-cyano-3,4-dihydro-2,2-dimethyl-2H-1 benzopyran-3-ol (Y-26763; an opener of ATP-sensitive K+ channels), and (R)-(+) trans-N-(4-pyridyl)-4-(1-aminoethyl)-cyclohexanecarboxamide (Y-27632; an inhibitor of Rho-associated kinase). Mean arterial pressure of the control group was 193+/-5 mm Hg (mean+/-S.E.M.), while that of the low-dose benidipine group was 183+/-5 mm Hg and that of the high-dose group was 159+/-4 mm Hg. Dilator responses of the basilar artery to acetylcholine and Y-26763 were impaired in SHR compared with those of normotensive Wistar-Kyoto (WKY) rats and treatment with benidipine enhanced the vasodilator responses to acetylcholine and Y-26763 in SHR. Y-27632-induced dilatation of the basilar artery was enhanced in SHR compared to that in WKY rats and the vasodilatation was reduced by benidipine in SHR. Sodium nitroprusside caused similar dilatation of the basilar artery, in both WKY rats and the SHR control group, and benidipine did not affect nitroprusside-induced dilatation of the artery in SHR. The wall of the basilar artery was significantly thicker in SHR than in WKY rats and benidipine treatment reduced the wall thickness of the artery in SHR. These findings suggest that chronic treatment with a Ca2+ channel blocker may enhance the dilator capacity and reduce contractility of the basilar artery during hypertension. Benidipine may also ameliorate the morphological changes of the basilar artery in hypertension. PMID- 11909608 TI - Enhancement of neurokinin A-induced smooth muscle contraction in human urinary bladder by mucosal removal and phosphoramidon: relationship to peptidase inhibition. AB - Neurokinin A (NKA) is potent in contracting the human detrusor muscle. Here, we have investigated whether these contractile responses are influenced by the presence of the mucosa, by the peptidase inhibitor phosphoramidon or by possible modulators, prostaglandins and nitric oxide. Contractile responses to neurokinin A were unaffected by indomethacin or N-omega-nitro-L-arginine, but were significantly reduced in strips containing mucosa. Phosphoramidon, an inhibitor of neutral endopeptidase 24.11 (neprilysin, CD10), was ineffective at 10 microM, but at 100 microM, significant increase in the maximum response was achieved by neurokinin A in detrusor strips with and without mucosa. In immunohistochemical studies, neutral endopeptidase immunoreactivity occurred in peripheral nerve trunks in the detrusor and in a fibrous meshwork in the subepithelial lamina propria. Our data indicate that neutral endopeptidase is present in bladder mucosa and detrusor, and support the concept that this metalloprotease and/or related enzymes are important in regulating the actions of tachykinins. PMID- 11909609 TI - Basolateral, but not apical, ATP inhibits vasopressin action in rat inner medullary collecting duct. AB - Previous studies have shown that basolateral ATP inhibits vasopressin action in the renal collecting tubule. Although there is evidence for an apical P2Y2 receptor in this tubule segment, it is not known whether apical ATP has similar effects. In the rat inner medullary collecting duct basolateral, but not apical, ATP (0.1-100 microM) reversibly inhibited vasopressin-induced increases in water permeability with an IC50 of 1.09 microM. Basolateral UTP, but not ADP, alpha,beta-methylene-ATP or 2-methylthio-ATP also inhibited vasopressin action. It is concluded that basolateral but not apical P2Y2 receptors inhibit vasopressin action in the collecting duct. PMID- 11909610 TI - Effects of CGS 21680, a selective adenosine A2A receptor agonist, on allergic airways inflammation in the rat. AB - We have investigated the effect of 2(4-((2-carboxymethyl)phenyl)ethylamino)-5'-N ethylcarboxamidoadenosine (CGS 21680), a potent and selective agonist at adenosine A2A receptors, on pulmonary inflammation induced by allergen challenge in the ovalbumin-sensitised, Brown Norway rat. Aerosol administration of ovalbumin (5 mg x ml(-1) for 60 min; calculated dose 0.4 mg x kg(-1)) induced increases in bronchoalveolar lavage fluid leukocyte numbers, protein content and myeloperoxidase and eosinophil peroxidase activities measured 24 h post challenge. CGS 21680 (10 and 100 microg x kg(-1) given intratracheally (i.t.) 30 min before and 3 h after allergen challenge) inhibited dose-dependently all the parameters of inflammation. Qualitatively similar results were obtained with the glucocorticosteroid, budesonide (0.1, 1 and 10 mg x kg(-1) given 3 h prior to ovalbumin challenge). CGS 21680 given i.t. reduced blood pressure in anaesthetised rats at similar doses to those at which anti-inflammatory effects were manifested. Both the anti-inflammatory and hypotensive responses to CGS 21680 were blocked by pretreatment with the selective adenosine A2A receptor antagonist, 4-(2-(7-amino-2-(2-furyl)(1,2,4)triazolo(2,3-a(1,3,5)triazin-5-yl amino)ethyl)phenol (ZM 241385), 3 mg x kg(-1) p.o., 1 h prior to the agonist. Thus, CGS 21680 manifests broad-spectrum anti-inflammatory activity in a model of allergic asthma in the Brown Norway rat through activation of adenosine A2A receptors. The striking similarity to budesonide, a clinically used anti inflammatory agent, suggests that adenosine A2A receptor agonists may be useful alternatives to glucocorticosteroids in the treatment of asthma. PMID- 11909611 TI - CX-659S: a novel diaminouracil derivative that has antioxidative and acute anti inflammatory activities. AB - We investigated the antioxidative activities and the effects on acute inflammation in mice of a novel diaminouracil derivative, CX-659S ((S)-6-amino-5 (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxamido)-3-methyl-1-phenyl-2,4(1H,3H) pyrimidinedione). CX-659S showed potent scavenging activities against the hydroxyl radical and peroxynitrite and inhibited lipid peroxidation in rat brain homogenates in vitro. Topically applied CX-659S dose-dependently inhibited arachidonic acid- and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear edema in mice. Consistent with its antioxidative properties in vitro, CX-659S dramatically attenuated the accumulation of lipid peroxides in the mouse ear elicited by repeated application of TPA. Previously, we reported the effectiveness of CX-659S against contact hypersensitivity reactions in both mouse and guinea pig models. These present results further suggest the therapeutic potential of CX-659S for acute skin inflammation that may involve oxidative tissue damage. PMID- 11909613 TI - The disappearing sample: researcher and research ability. PMID- 11909612 TI - Hyperglycemia induced by intracerebroventricular choline: involvement of the sympatho-adrenal system. AB - Intracerebroventricular (i.c.v.) injection of choline (75-300 microg) produced a dose-dependent increase in blood glucose levels. Pre-treatment with the nicotinic acetylcholine receptor antagonist, mecamylamine (50 microg, i.c.v.) blocked the hyperglycemia induced by choline (150 microg, i.c.v.), but the response was not affected by pre-treatment with the muscarinic acetylcholine receptor antagonist, atropine (10 microg, i.c.v.). Pre-treatment with the neuronal choline uptake inhibitor, hemicholinium-3 (20 microg, i.c.v.), attenuated the hyperglycemia induced by choline. The hyperglycemic response to choline was associated increased plasma levels of adrenaline and noradrenaline. The hyperglycemia elicited by choline was greatly attenuated by bilateral adrenalectomy, and entirely blocked by either surgical transection of the splanchnic nerves or by pre-treatment with the alpha-adrenoceptor antagonist, phentolamine. These data show that choline, a precursor of acetylcholine, increases blood glucose and this effect is mediated by central nicotinic acetylcholine receptor activation. An increase in sympatho-adrenal activity appears to be involved in the hyperglycemic effect of choline. PMID- 11909614 TI - Supporting family carers through the use of information and communication technology--the EU project ACTION. AB - Assisting Carers Using Telematics Interventions to meet Older persons' Needs (ACTION) is an EU project designed to improve the quality of life of frail older and disabled people and their family carers by the use of information and communication technology. It involved Sweden, England, Northern Ireland, the Republic of Ireland and Portugal. This article provides an overview of the research and development process of the ACTION system and services. The focus is upon the evaluation results with regard to the quality of life of older people and their family carers, the usability of ACTION and cost considerations. Recommendations are made regarding user-focused technology such as ACTION. PMID- 11909615 TI - Coping and adjustment in Chinese patients with chronic obstructive pulmonary disease. AB - The purpose of this study was to examine the relationships between coping and psychosocial adjustment of Chinese patients with chronic obstructive pulmonary disease (COPD). A convenience sample of 54 hospitalized COPD adult patients participated in the study. All participants had moderate to severe scores in respect of pulmonary functional status and symptoms. The findings indicated that the participants adopted limited coping strategies and had poor psychosocial adjustment to their illness. No significant correlation was found between the total scores for coping and psychosocial adjustment. Both confrontive and optimistic coping scores of the perceived use and helpfulness of the coping scale were negatively correlated with the total adjustment score. Fatigue, older age, worse perception of current health status, and use of confrontive and emotive coping styles were predictors of worse psychosocial adjustment. This study provides a better understanding of the needs and difficulties experienced by Chinese COPD patients and concludes that modification of such factors by health professionals may lead to a better psychosocial adjustment. PMID- 11909616 TI - Seeing need and developing care: exploring knowledge for and from practice. AB - The use and creation of knowledge by practitioners is complex. There is a contemporary emphasis on the implementation of knowledge, or evidence, derived from outside the immediate practice environment. However, there is also an appreciation of the context-specific nature of practitioner knowledge that they use to inform patient care, and that is derived from their intimate contact with patients. This paper described one study that sought to analyse the impact of developments in practice on the wider professional and organisational community. Three case study sites were identified through a multidimensional sampling matrix, and interviews conducted with 41 practitioners of various organisational positions and professions. The primacy of context-specific, or proximal knowledge was highlighted, together with a manipulation of evidence and policy to fit understandings of local patient need. PMID- 11909617 TI - Managing technology in the intensive care unit: the nurses' experience. AB - This study investigates the experiences of a group of critical care nurses regarding the use of technology in the intensive care unit. The study is grounded in the phenomenological hermeneutics of Martin Heidegger and argues that this methodology is compatible with the humanistic values of nursing. Data were generated from in-depth interviews and overt participant observation. The process of data analysis was guided by Heidegger's notions of phenomenological reflection and hermeneutic circle, (Van Manen's, Researching lived experience: human science for an action sensitive pedagogy. The State University of New York press, Ontario, Canada, 1990) process of thematic analysis and (Draper's, Nursing perspectives on quality of life. Routledge, London, 1997) analytical principles. The ability to manage the technology emerged as a main component of being a critical care nurse. It is revealed also that the nurses' ability to manage the technology is gained mainly through experience. The effect of machinery management on patient care is seen as part of everyday routine in the critical care setting. Moreover, technical activities are seen as more important and stimulating than other nursing activities. PMID- 11909619 TI - Psychosocial factors influencing nurses' involvement with organ and tissue donation. AB - This paper focuses on the first phase of a 3-year study that explored the psychosocial factors that influence nurses' willingness to discuss post-mortem donation intentions with relatives of potential organ and tissue donors. The United Kingdom's donation system is dependent upon such discussions taking place. A cross-sectional survey of 776 randomly selected nurses, from two health regions in the United Kingdom, found that personal negative attitudes to aspects of donation and transplantation, fears and misconceptions about the donation process, clinical area of work, past experience, and socio-historical factors influence discussion behaviour. Knowledge deficits were discovered, together with requests for general information about the donation process and specific information about the organ and tissue donor exclusion criteria. PMID- 11909618 TI - Fear and in-hospital social support for coronary artery bypass grafting patients on the day before surgery. AB - The purpose of this study was to ascertain the amount of in-hospital social support received by coronary artery bypass grafting patients and the impact of this support on their feelings of fear and anxiety. As adapted from Kahn's theory, social support was understood as emotional, informational and tangible support. The bypass grafting fear scale was developed to measure the fear, and the hospital anxiety and depression scale and the state anxiety inventory were used to measure the anxiety. Data were collected pre-operatively with a questionnaire from in-patients (N=193) and analysed using logistic regression analysis and one-way ANOVA. The majority of patients received plenty of social support from nurses and a great deal of multiprofessional counselling. When the amount of social support was high, patients experienced lower levels of fear and anxiety. It is concluded that social support from nurses can effectively reduce pre-operative fear and anxiety, but that the amount of support should be high. PMID- 11909620 TI - The nurse specialist as main care-provider for patients with type 2 diabetes in a primary care setting: effects on patient outcomes. AB - A solution to safeguard high quality diabetes care may be to allocate care to the nurse specialist. By using a one group pretest-posttest design with additional comparisons, this study evaluated effects on patient outcomes of a shared care model with the diabetes nurse as main care-provider for patients with type 2 diabetes in a primary care setting. The shared care model resulted in an improved glycaemic control, additional consultations and other outcomes being equivalent to diabetes care before introduction, with the general practitioner as main care provider. Assignment of care for patients with type 2 diabetes to nurse specialists seems to be justified. PMID- 11909621 TI - Coping with organizational stress among hospital nurses in Southern Ontario. AB - Government cutbacks and anticipated staff reductions were hypothesized to be a unique source of organizational stress. The study focused on how nurses coped with stress and whether any strategy effectively reduced occupational stress. A sample of 107 nurses were asked to rate their occupational stress, job satisfaction, and coping strategies. Avoidance and social support were found to be significantly correlated with stress, but neither of these coping strategies appeared to reduce nurses' level of organizational stress. However, an interaction between problem solving and job satisfaction was found to be highly significant and it added 42% to predicting stress levels. Supporting the stress buffering hypothesis, nurses with lower intrinsic job satisfaction seemed to benefit from employing problem solving as a coping strategy whereas dissatisfied nurses who infrequently use problem solving reported the highest levels of organizational stress. Paradoxically, intrinsically satisfied nurses who most frequently utilize problem solving experienced heightened organizational stress. PMID- 11909622 TI - Interventions used by nursing staff members with psychogeriatric patients resisting care. AB - Although resistance to care can have a major impact on the provision of care, relatively limited research has been reported on the topic. This research examined differences in the use of interventions by nurses in different care settings to manage resistance to care with eating and dressing. A convenience sample of 50 nurses (34 working in psychiatric hospitals and 16 in nursing homes) participated in the study. Nurses in both settings reported using similar interventions for both problems. Non-nursing interventions were consistently mentioned to be more common when resistance was accompanied by physical aggression than by verbal aggression. PMID- 11909623 TI - Violence against nurses in healthcare facilities in Kuwait. AB - A national cross-sectional survey was conducted to document the prevalence and determinants of violence against nurses in healthcare facilities in Kuwait. It involved all nurses employed in all types of health-related facilities and available in the country in May 1999. The questionnaire was completed by 5876 nurses (85% females, 88% non-Kuwaitis). Verbal violence had been experienced in the 6 previous months by 48% of the group, and physical violence by 7%. There was no physical harm reported in 63% of cases of physical violence. Physical abusers were mostly patients (51%). Compared to nurses who had never experienced physical violence, those who had experienced some were more likely to be male, non Kuwaiti, to have had a shorter professional experience, and to be working in a hospital rather than in a primary healthcare center. The experience of nurses with violence is still relatively rare in Kuwait. Communication with patients and their entourage of family members and/or close friends is needed to clarify expectations and to avoid frustration and angry verbal outbursts. PMID- 11909624 TI - A beta-glucosidase/xylosidase from the phytopathogenic oomycete, Phytophthora infestans. AB - An 85-kDa beta-glucosidase/xylosidase (BGX1) was purified from the axenically grown phytopathogenic oomycete, Phytophthora infestans. The bgx1 gene encodes a predicted 61-kDa protein product which, upon removal of a 21 amino acid leader peptide, accumulates in the apoplastic space. Extensive N-mannosylation accounts for part of the observed molecular mass difference. BGX1 belongs to family 30 of the glycoside hydrolases and is the first such oomycete enzyme deposited in public databases. The bgx1 gene was found in various Phytophthora species, but is apparently absent in species of the related genus, Pythium. Despite significant sequence similarity to human and murine lysosomal glucosylceramidases, BGX1 demonstrated neither glucocerebroside nor galactocerebroside-hydrolyzing activity. The native enzyme exhibited glucohydrolytic activity towards 4 methylumbelliferyl (4-MU) beta-D-glucopyranoside and, to lesser extent, towards 4 MU-D-xylopyranoside, but not towards 4-MU-beta-D-glucopyranoside. BGX1 did not hydrolyze carboxymethyl cellulose, cellotetraose, chitosan or xylan, suggesting high substrate specificity and/or specific cofactor requirements for enzymatic activity. PMID- 11909625 TI - Alkaloid production in Duboisia hybrid hairy root cultures overexpressing the pmt gene. AB - Putrescine:SAM N-methyltransferase (PMT) catalyses the N-methylation of the diamine putrescine to form N-methylputrescine, the first specific precursor of both tropane and pyridine-type alkaloids, which are present together in the roots of Duboisia plants. The pmt gene of Nicotiana tabacum was placed under the regulation of the CaMV 35S promoter and introduced into the genome of a scopolamine-rich Duboisia hybrid by a binary vector system using the disarmed Agrobacterium tumefaciens strain C58C1 carrying the rooting plasmid pRiA4. The presence of the foreign gene in kanamycin-resistant hairy roots and its overexpression were confirmed by polymerase chain reaction and Northern blot analysis respectively. The N-methylputrescine levels of the resulting engineered hairy roots increased (2-4-fold) compared to wild type roots, but there was no significant increase in either tropane or pyridine-type alkaloids. PMID- 11909626 TI - Changes in cell wall polysaccharides of Silene vulgaris callus during culture. AB - In Silene vulgaris (M.) G. cell culture three growth phases were distinguished, namely, a lag phase, an exponential phase and a stationary phase. Pectin termed silenan and an acidic arabinogalactan were isolated as cell wall polysaccharides of S. vulgaris callus at the different growth phases during culture. Production of silenan as the galacturonan (or rhamnogalacturonan) core was observed at the beginning of the exponential phase and at the stationary phase of the callus growth. Arabinogalactan, containing the galacturonic acid residues, is formed at the exponential phase followed by attachment to the core of silenan in the middle of the exponential phase. The arabinogalactan constituent of silenan appeared to be destroyed gradually at the stationary growth phase. The monosaccharide compositions of silenan and arabinogalactan were determined at various phases of the callus growth. Silenan was found to be formed in maximum amounts at the exponential phase of the cell growth. Insignificant alterations of the yields of acidic arabinogalactan were found during culture while total productivity per litre of medium and rate of production per day of arabinogalactan were found to be maximal at the exponential phase of growth. PMID- 11909627 TI - Monoterpenoid accumulation in Melaleuca alternifolia seedlings. AB - Individual leaves of the commercial terpinen-4-ol type of Melaleuca alternifolia were examined both quantitatively and qualitatively for volatile constituents from the emergence of the first true leaves, through to 6-week-old tenth leaf set material. A GC internal standard addition method was used to measure changes in oil composition and the accumulation of volatile constituents expressed on a dry weight, unit leaf area and whole leaf basis. In the early stages of seedling growth, leaves contained higher concentrations of terpinolene, alpha-pinene and beta-pinene and lower concentrations of terpinen-4-ol, sabinene and cis-sabinene hydrate than mature leaf. Concentrations of the former constituents fell and the latter rose by the time leaf set 10 was 6 weeks old. Key constituent, 1,8-cineole remained in similar concentration throughout ontogeny. The variation in concentration of other key constituents during early stages of seedling development suggests that caution is required in extrapolating seedling leaf data to mature tree oil quality. PMID- 11909628 TI - Survey of aliphatic glucosinolates in Sicilian wild and cultivated Brassicaceae. AB - In the frame of the activities carried out to exploit Sicilian local cultivars of brassicas, we focused our attention on some of the potential health compounds of various local cruciferous crops. These compounds are of interest to improve the quality of the produce with the aim to develop new cultivars capable of providing functional foods able to prevent disease. In this context, we surveyed for the presence of specific glucosinolates in local cultivars of broccoli, cauliflower, kale, and in some wild species widespread in Sicily, using as control various commercial cultivars. Glucosinolate composition varied extensively among species and crops of the same species, such as cauliflower, broccoli and kale. Cultivar variation for glucosinolate profile was also observed for some crops. For example, Sicilian cultivars of cauliflower possessing colored curds displayed a high content of glucosinolates, glucoraphanin in particular, compared to white curd commercial cultivars. Also some wild species had a high content of other glucosinolates. PMID- 11909630 TI - Neogrifolin derivatives possessing anti-oxidative activity from the mushroom Albatrellus ovinus. AB - Three neogrifolin derivatives, 3-hydroxyneogrifolin, 1-formylneogrifolin and 1 formyl-3-hydroxyneogrifolin along with grifolin and neogrifolin were isolated from the Japanese mushroom Albatrellus ovinus belonging to Scutigeraceae. Their structures were established by a combination of two-dimensional NMR spectroscopic analyses and by chemical synthesis. 3-Hydroxyneogrifolin and 1-formyl-3-hydroxy neogrifolin showed more potent antioxidative activity properties than either alpha-tocopherol or BHA. PMID- 11909629 TI - Production of taxoids with biological activity by plants and callus culture from selected Taxus genotypes. AB - Twenty seven different yew trees belonging to various genotypes and hybrids have been screened for their capacity to produce significant amounts of taxoids provided with biological activity in the tubulin test. From the three best genotypes selected, Taxus x media "Sargentii" proved to be able to produce viable calluses from excised roots placed in vitro. Taxoid composition at various times of the in vitro culture was determined and the carcinostatic efficiency of the extracts was established using the KB cell cytotoxicity test. In leaves and calluses, respectively, 0.069 and 0.032% paclitaxel (taxol) contents were found. These contents were significantly higher than those previously reported for other genotypes. PMID- 11909631 TI - Pyrano-isoflavones with erectile-dysfunction activity from Eriosema kraussianum. AB - Five pyrano-isoflavones have been isolated from the rootstock of Eriosema kraussianum N. E. Br (Papilionaceae). Spectral data and single crystal X-ray analyses were used for structural elucidation. The most active of the compounds had an activity of 75% of that found in Viagra in the erectile dysfunction test on rabbit penile smooth muscle. PMID- 11909632 TI - Proanthocyanidin glycosides and related polyphenols from cacao liquor and their antioxidant effects. AB - Purification of polar fractions from cacao liquor extracts gave 17 phenolics including four new compounds. The new compounds were characterized as a C glycosidic flavan, an O-glycoside of a dimeric and two O-glycosides of trimeric A linked proanthocyanidins, on the basis of spectroscopic data. Isolated polyphenols showed inhibitory effects on nicotinamide adenine dinucleotide phosphate-dependent lipid peroxidation in microsomes and on the autoxidation of linoleic acid. These effects were attributed to the radical-scavenging activity in the peroxidation chain reactions, based on the findings that the cacao polyphenols effectively scavenged the 1,1-diphenyl-2-picrylhydrazyl radical. PMID- 11909633 TI - Characterization of betaines using electrospray MS/MS. AB - Betaines are an important class of naturally occurring compounds that function as compatible solutes or osmoprotectants. Because of the permanent positive charge on the quaternary ammonium moiety, mass spectrometric analysis has been approached by desorption methods, including fast atom bombardment and plasma desorption mass spectrometry. Here we show that electrospray ionization MS gives comparable results to plasma desorption MS for a range of authentic betaine standards and betaines purified from plant extracts by ion exchange chromatography. A distinct advantage of electrospray ionization MS over plasma desorption MS is the capability of obtaining product ion spectra via MS/MS of selected parent ions, and hence structural information to discriminate between ions of identical mass. PMID- 11909634 TI - Sesquiterpene pyridine alkaloids from Hippocratea excelsa. AB - Nineteen sesquiterpene pyridine alkaloids including 17 new compounds have been isolated from the 70% aq. EtOH extract of stem barks of Hippocratea excelsa. The structures of these compounds were elucidated by various spectroscopic means. PMID- 11909635 TI - Secoiridoid and iridoid glucosides from Syringa afghanica. AB - Phytochemical investigation of the dried leaves of Syringa afghanica, has led to the isolation of nine secoiridoid glucosides, safghanosides A-H and 2"-epi frameroside, as well as an iridoid glucoside, syringafghanoside along with nineteen known compounds. The structures were elucidated by spectroscopic and chemical means. PMID- 11909637 TI - Toll-like receptors as adjuvant receptors. AB - The mammalian Toll-like receptors (TLRs) are expressed on macrophages and dendritic cells, which are primarily involved in innate immunity. At present, ligands for several of the TLRs, such as TLR2, TLR3, TLR4, TLR5, TLR6, and TLR9, have been identified. Most of these ligands are derived from pathogens, but not found in the host, suggesting that the TLRs are critical to sensing invading microorganisms. Pathogen recognition by TLRs provokes rapid activation of innate immunity by inducing production of proinflammatory cytokines and upregulation of costimulatory molecules. Activated innate immunity subsequently leads to effective adaptive immunity. In this regard, the TLRs are considered to be adjuvant receptors. Distinct TLRs can exert distinct, but overlapping sets of biological effects. Accumulating evidence indicates that this can be attributed to both the common and unique aspects of the signaling mechanisms that mediate TLR family responses. For example, TLR2 and TLR9 require MyD88 as an essential signal transducer, whereas TLR4 can induce costimulatory molecule upregulation in a MyD88-independent manner. Understanding the TLR system should offer invaluable opportunity for manipulating host immune responses. PMID- 11909638 TI - Upregulation by retinoic acid of transforming growth factor-beta-stimulated heat shock protein 27 induction in osteoblasts: involvement of mitogen-activated protein kinases. AB - We investigated whether transforming growth factor-beta (TGF-beta) stimulates the induction of heat shock protein (HSP) 27 and HSP70 in osteoblast-like MC3T3-E1 cells and the mechanism underlying the induction. TGF-beta increased the level of HSP27 but had no effect on the HSP70 level. TGF-beta stimulated the accumulation of HSP27 dose-dependently, and induced an increase in the level of mRNA for HSP27. TGF-beta induced the phosphorylation of p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase. The HSP27 accumulation induced by TGF-beta was significantly suppressed by PD98059, an inhibitor of the upstream kinase of p44/p42 MAP kinase, or SB203580, an inhibitor of p38 MAP kinase. PD98059 and SB203580 suppressed the TGF-beta-stimulated increase in the level of mRNA for HSP27. Retinoic acid, a vitamin A (retinol) metabolite, which alone had little effect on the HSP27 level, markedly enhanced the HSP27 accumulation stimulated by TGF-beta. Retinoic acid enhanced the TGF-beta-induced increase of mRNA for HSP27. The amplification of TGF-beta-stimulated HSP27 accumulation by retinoic acid was reduced by PD98059 or SB203580. Retinoic acid failed to affect the TGF-beta induced phosphorylation of p44/p42 MAP kinase or p38 MAP kinase. These results strongly suggest that p44/p42 MAP kinase and p38 MAP kinase take part in the pathways of the TGF-beta-stimulated HSP27 induction in osteoblasts, and that retinoic acid upregulates the TGF-beta-stimulated HSP27 induction at a point downstream from p44/p42 MAP kinase and p38 MAP kinase. PMID- 11909639 TI - Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19 oxidoprogesterone. AB - The alkylation of amino groups of the mineralocorticoid receptor (MR) with pyridoxal 5'-phosphate or 2,4,6-trinitrobenzenesulphonate (TNBS) under controlled conditions modifies only one lysyl residue, which accounts for a 70% inhibition of steroid binding capacity. The Kd of aldosterone for MR is not affected by the treatment, but the total number of binding sites is greatly decreased. The modified receptor is capable of dynamically conserving its association with the hsp90-based heterocomplex. Importantly, the binding of natural agonists protects the hormone binding capacity of the MR from the inactivating action of alkylating agents. In contrast, antagonistic steroids are totally incapable of providing such protection. Like the antagonistic ligands, and despite its potent mineralocorticoid biological effect, the sole MR specific synthetic agonist known to date, 11,19-oxidoprogesterone (11-OP), shows no protective effect upon treatment of the MR with pyridoxal 5'-phosphate or TNBS. Limited digestion of the MR with alpha-chymotrypsin generates a 34 kDa fragment, which becomes totally resistant to digestion upon binding of natural agonists, but not upon binding of antagonists. Interestingly, the synthetic 21-deoxypregnanesteroid 11-OP exhibits an intermediate pattern of proteolytic degradation, suggesting that the conformational change generated in the MR is not equivalent to that induced by antagonists or natural agonists. We conclude that in the first steps of activation, the MR changes its conformation upon binding of the ligand. However, the nature of this conformational change depends on the nature of the ligand. The experimental evidence shown in this work suggests that a single lysyl group can determine the hormone specificity of the MR. PMID- 11909640 TI - Modulation of intrinsic P-glycoprotein expression in multicellular prostate tumor spheroids by cell cycle inhibitors. AB - The effects of cell cycle inhibition on the expression of the multidrug resistance transporter P-glycoprotein (P-gp) as well as of the cyclin-dependent kinase (CDK) inhibitors p27(Kip1) and p21(WAF-1) were investigated in DU-145 prostate tumor spheroids. With increasing spheroid size the number of cells in the G0/G1 phase augmented, whereas the number of cells in the G2/M phase and the S phase of the cell cycle declined. The number of G0/G1 cells was elevated after incubation with either mimosine, staurosporine or serum-free medium. Mitomycin C and roscovitine increased the number of S phase cells. Roscovitine additionally increased cells in the G2/M phase. Incubation in serum-free medium upregulated p21(WAF-1), p27(Kip1) and P-gp. Mimosine treatment resulted in upregulation of p27(Kip1) and P-gp, whereas p21(WAF-1) remained unchanged. Upon roscovitine treatment p27(Kip1) and p21(WAF-1) were downregulated, whereas P-gp was unaltered. Mitomycin C treatment resulted in downregulation of p27(Kip1) and p21(WAF-1); no significant change in P-gp levels was observed. Staurosporine induced upregulation of p21(WAF-1) whereas p27(Kip1) remained unaltered. P-gp was downregulated upon staurosporine treatment, which was owing to an elevation of intracellular reactive oxygen species by this compound. It is concluded that upregulation of P-gp in G0/G1 phase cells requires coexpression of the CDK inhibitor p27(Kip1) but not the CDK inhibitor p21(WAF-1). PMID- 11909641 TI - Cytotoxic effects of metal complexes of biogenic polyamines. I. Platinum(II) spermidine compounds: prediction of their antitumour activity. AB - Cytotoxicity and cell growth inhibition studies were performed for three distinct polynuclear platinum(II) complexes of spermidine, which showed to have significant cytotoxic and antiproliferative properties on the HeLa cancer cell line. The chemical environment of the metal centres in the drugs, as well as the coordination pattern of the ligand, were found to be strongly determinant of their cytotoxic ability. In the light of the results gathered, the most effective anticancer compound among the ones tested (IC50=5 microM) was found to be the one displaying three difunctional (PtCl2N2) moieties ((PtCl2)3(spd)2). Both the cytotoxic activity and the antiproliferative properties of the complexes studied showed to be irreversible for all the concentrations tested. PMID- 11909642 TI - Identification of interaction between MEK2 and A-Raf-1. AB - Mitogen-activated protein (MAP) kinases are activated by dual-specificity kinases, termed MEKs. Using MEK2 as bait in yeast two-hybrid screening, besides c Raf and KSR, A-Raf was identified as a novel partner that interacts with MEK2. This interaction was confirmed by in vitro binding assay. Further investigation indicates that regions critical for this interaction were located between residues 255 and 606 that represent the kinase domain of A-Raf. PMID- 11909643 TI - Activation of NKCC1 by hyperosmotic stress in human tracheal epithelial cells involves PKC-delta and ERK. AB - Hyperosmotic stress activates Na+-K+-2Cl- cotransport (NKCC1) in secretory epithelia of the airways. NKCC1 activation was studied as uptake of 36Cl or 86Rb in human tracheal epithelial cells (HTEC). Application of hypertonic sucrose or NaCl increased bumetanide-sensitive ion uptake but did not affect Na+/H+ and Cl /OH-(HCO3-) exchange carriers. Hyperosmolarity decreased intracellular volume (Vi) after 10 min from 7.8 to 5.4 microl/mg protein and increased intracellular Cl- (Cl-i) from 353 to 532 nmol/mg protein. Treatment with an alpha-adrenergic agent rapidly increased Cl-i and Vi in a bumetanide-sensitive manner, indicating uptake of ions by NKCC1 followed by osmotically obligated water. These results indicate that HTEC act as osmometers but lose intracellular water slowly. Hyperosmotic stress also increased the activity of PKC-delta and of the extracellular signal-regulated kinase ERK subgroup of the MAPK family. Activity of stress-activated protein kinase JNK was not affected by hyperosmolarity. PD 98059, an inhibitor of the ERK cascade, reduced ERK activity and bumetanide sensitive 36Cl uptake. PKC inhibitors blocked activation of ERK indicating that PKC may be a downstream activator of ERK. The results indicate that hyperosmotic stress activates NKCC1 and this activation is regulated by PKC-delta and ERK. PMID- 11909645 TI - Cortical connectivity in high frequency beta-rhythm in schizophrenics with positive and negative symptoms. AB - During the last decade the role of high frequency EEG activity in the 'binding phenomenon' was discovered. It was supposed that this phenomenon provided the integration between different brain structures underlying higher nervous functions and possibly even consciousness [Proc. Natl. Acad. Sci. 90 (1993) 2078; Annu. Rev. Neurosci. 18 (1995) 555; J. Neurosci. V 16 (1996) 4240; Am. Physiol. Soc. (1998) 1567; Induced Rhythms in the Brain (1992) 425; NeuroReport 8 (1997) 531; Proc. Natl. Acad. Sci. USA 94 (1997) 12198]. Schizophrenia is considered as a disorder of the integration between different brain regions [Review of Psychiatry 18 (1999a) 29; Conceptual Advances in Russian Neuroscience: Complex Brain Functions (1999) 151; Brain Res. Rev. 31 (2000) 301], and in the present work we have studied cortical connectivity, focusing on those connections which are maintained by high frequency EEG-rhythm (20-40 Hz). The results showed a high degree of biopotential synchronisation between definite cortical areas during cognitive processes in normal subjects and have evidenced significant functional connectivity disturbances in schizophrenia in this EEG frequency domain. PMID- 11909644 TI - Induced gamma activity is associated with conscious awareness of pattern masked nouns. AB - The aim of this study was to explore the relationship between electroencephalographic (EEG) activity in the gamma frequency range and conscious awareness of a visual stimulus. EEG was recorded from subjects while they were shown backward-masked words only some of which they were able to discriminate correctly. The results showed that activity in the gamma frequency range increased with reported awareness of a word independently of whether it was correctly discriminated or not. It is concluded that gamma power is associated with awareness-dependent visual processing but not with processing that occurs in the absence of awareness. PMID- 11909646 TI - Frontal and parieto-temporal cortical ablations and diaschisis-like effects on auditory neurocognitive potentials evocable from apparently intact ipsilateral association areas in humans: five case reports. AB - The aim of this study was to investigate the effects of disruption on the warning auditory S1-elicited ERP and CNV complexes recordable on the site and on remote ipsilateral apparently normal anatomo-functionally interconnected brain regions. These effects in some cases showed aspects of a probable diaschisis-like phenomenon, due to resections of extensive frontal association cortex or of primary and secondary sensory parieto-temporal areas damaged by differing pathological processes. Using a standard CNV paradigm, 21/19 EEG electrodes connected with three different references, and scalp-topographic bidimensional mapping analysis, the S1 auditory binaural/monaural clicks N1a,b,c, P2, N2, P3 and CNV waves were recorded in 10 normal subjects and 11 patients. Nine of the latter had been submitted to unilateral frontal dorsolateral cortex ablation, one to bihemispheric dorsomedial cortex ablation, and one to unilateral ablation of sensory parieto-temporal cortex and underlying white matter, verified through CT/MRI examinations. No true S1ERP/CNV components were recordable over the ablated cortical areas, whereas normal ERP/CNV complexes were observable on the intact hemispheres. In five patients, four of whom with frontocortical ablations, the S1 ERP/CNV complexes appeared severely diminished or disrupted, in two cases in a slow, partially-reversible manner, also in the neuroradiologically normal ipsilateral functionally-connected post-rolandic sensory and association areas. Similar deactivation of some ERP components was observed in reverse on the unilateral dorsolateral frontocortical region in the fifth patient with parieto temporal cortex ablation. Even when they are partially reversible, these ipsilateral remote ERP changes in apparently intact brain regions, due to ablations of functionally-interconnected cortical formations, probably reflect cortical deactivation or simply dysfacilitation deriving from functional unilateral diaschisis. If these changes are instead irreversible they may probably be interpreted as transneuronal degeneration phenomena, though they are not at present easy to document either neuroradiologically or electroclinically. PMID- 11909647 TI - Phase-coupling of theta-gamma EEG rhythms during short-term memory processing. AB - Because of the importance of oscillations as a general phenomenon of neuronal activity the use of EEG spectral analysis is among the most important approaches for studying human information processing. Usually, oscillations at different frequencies occur simultaneously during information processing. Thus, the question for synchronisation of different frequencies by phase coupling and its possible functional significance is of primary importance. An answer may be given by bispectral analysis. Estimation of the (cross-) bispectrum allows to identify synchronised frequencies and possibly, the existence of non-linear phase coupling of different oscillators. Previous studies have demonstrated the simultaneous occurrence of slow (4-7 Hz) and fast (20-30 Hz) oscillations at frontal and prefrontal electrode positions during memory processing. However, interrelations between these rhythms have not been investigated up to now. In order to test short-term memory, the Sternberg task with random figures and number words was carried out on 10 female subjects. During the task EEG was recorded. Power and bispectral analyses from frontal, prefrontal and frontopolar regions were performed off-line. Increased power was found in both the theta and the gamma bands. Strong phase-coupling between theta at Fz and gamma at F3 and at Fp1, respectively, was shown for memorising number words by means of cross bicoherence. A possible reason for this is an amplitude modulation of gamma frequencies by slow oscillations. The correspondent coherence analysis between the envelope of gamma frequencies at Fp1 and the raw EEG at Fz supports this presumption. This finding is interpreted as an EEG aspect of the functional linking between the prefrontal areas and the G.cinguli (as part of the limbic system), which are both extremely important for memory functions. PMID- 11909648 TI - Appraisals and impression management opportunities: person and situation influences on cardiovascular reactivity. AB - This study examined how threat and coping appraisals and impression management opportunities influenced cardiovascular reactivity within a self-presentation context. Participants were videotaped performing a speech that they believed might be evaluated. Participants' physiological responses were recorded 1 week later while: (a) watching their speech, indicating which portions they wished to re-shoot before it was evaluated,;(b) simply watching their speech; or (c) watching an architectural videotape and indicating which portions should be reshot, and also watching their own speech, but without evaluation concerns. Cardiovascular reactivity was influenced by the presence or absence of the impression management opportunity, but more variance in reactivity was explained when participants' threat and coping appraisals were also taken into account. The specific cardiovascular responses pertinent to goal-relevant action within each condition were the only responses related to appraisals. These findings emphasize the importance of considering both the situation and the state of the individual in that situation when attempting to understand social psychological influences on physiological reactivity. PMID- 11909649 TI - Musical beat influences corticospinal drive to ankle flexor and extensor muscles in man. AB - Body movements in man are frequently observed in relation to musical rhythms. In this study we have investigated the effect of strongly rhythmic music on corticospinal drive to the ankle extensor and flexor muscles involved in foot tapping. Surface electromyographic (EMG) recordings were made from tibialis anterior (TA) and lateral gastrocnemius (LGN) muscles in 12 normal subjects. Rock music with a strong rhythmic beat or white noise was played. Transcranial magnetic stimulation (TMS) was delivered to the motor cortex in time with the music to produce motor evoked potentials (MEPs) in both muscles while relaxed. During white noise trials nine subjects showed a significant correlation of MEP areas in TA with LGN, compared with only three subjects during music. Overall, there was a significant decrease in the correlation coefficient during music. We conclude that correlated variations in MEP areas between the muscles seen during white noise can be destroyed in the presence of music. This may be due to sub threshold variations in corticospinal excitability to ankle extensors and flexors, which are time-locked to the musical beat but out of phase with one another. PMID- 11909650 TI - A review of Anglo-American forensic professional codes of ethics with considerations for code design. AB - Continuing instances of ethical misconduct and recent advances in the practice of forensic science, the latter reflected by increased growth and professionalism, along with heightening public scrutiny, require that the profession be clear, current, and forthright about its core values. A way of expressing these values is through the codes of ethics of its professional membership organizations. Codes of ethics/conduct from 14 Anglo-American professional forensic membership organizations are reviewed for their clarity, construction, cogency, and the normative form of their provisions. Ethical precepts from the forensic codes reviewed are organized, in summary fashion, by obligation form and application context. Functions that professional codes have served in other professions are described. The codes are found to vary considerably. Some provisions are unclear, and most of the codes are less than comprehensive. There is very little guidance for application of ethical obligations in any of the codes reviewed. Suggestions for redesign, with a partial example, are provided. PMID- 11909651 TI - Simultaneous analysis of gamma-hydroxybutyric acid, gamma-butyrolactone, and 1,4 butanediol by micellar electrokinetic chromatography. AB - A micellar electrokinetic chromatography (MEKC) method was optimised for simultaneous analysis of gamma-hydroxybutyric acid (GHB), gamma-butyrolactone (GBL), and 1,4-butanediol (BD). Best conditions for separation and baseline stability were achieved using a carrier electrolyte comprising 30.0mM sodium barbital and 150.0mM sodium dodecyl sulphate (SDS) at pH 10.2. Calibration functions were linear, giving correlation coefficients (r(2)) >0.998 for the three target compounds. Limits of detection (LOD) defined as three times the noise, were 5.1mg/l, 0.34 and 0.25g/l for GHB, GBL and BD, respectively. The repeatability of migration times and peak areas, expressed as the R.S.D. (n = 9) was better than 0.41 and 3.05%, respectively. Some casework samples were analysed using the optimised conditions. PMID- 11909652 TI - Quantitative determination of amphetamines, cocaine, and opiates in human hair by gas chromatography/mass spectrometry. AB - Hair of young subjects (N = 36) suspected for drug abuse was analysed for morphine, codeine, heroin, 6-acetylmorphine, cocaine, methadone, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4 methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA). The analysis of morphine, codeine, heroin, 6-acetylmorphine, cocaine, and methadone in hair included incubation in methanol, solid-phase extraction, derivatisation by the mixture of propionic acid anhydride and pyridine, and gas chromatography/mass spectrometry (GC/MS). For amphetamine, methamphetamine, MDA, MDMA, and MDEA analysis, hair samples were incubated in 1M sodium hydroxide, extracted with ethyl acetate, derivatised with heptafluorobutyric acid anhydride (HFBA), and assayed by GC/MS. The methods were reproducible (R.S.D. = 5.0-16.1%), accurate (85.1-100.6%), and sensitive (LoD = 0.05-0.30ng/mg). The applied methods confirmed consumption of heroin in 18 subjects based on positive 6 acetylmorphine. Among these 18 heroin consumers, methadone was found in four, MDMA in two, and cocaine in two subjects. Cocaine only was present in two, methadone only in two, methamphetamine only in two, and MDMA only in seven of the 36 subjects. In two out of nine coloured and bleached hair samples, no drug was found. Despite the small number of subjects, this study has been able to indicate the trend in drug abuse among young people in Croatia. PMID- 11909653 TI - HPLC simultaneous determination of glycerol and mannitol in human tissues for forensic analysis. AB - A method for simultaneous determination of glycerol and mannitol in various human tissues was devised and for this we used high-performance liquid chromatography (HPLC). Specimens were homogenized in a mixture of chloroform and methanol, phosphate buffer (pH 7.0) and pentaerythritol (IS) solution. After centrifugation, an aliquot of the aqueous layer was evaporated to dryness and derivatized with p-nitrobenzoyl chloride at 50 degrees C for 1h, then applied to HPLC with analytical conditions of: column, CAPCELL PAK C18 MG (250 mm x 3.0 mm i.d., 5 microm, Shiseido Co. Ltd., Tokyo, Japan); column temperature, 1-2 degrees C; mobile phase, 75% acetonitrile-distilled water containing 0.05% trifluoroacetic acid, 0.05% heptafluoro-n-butyric acid and 0.1% triethylamine; flow rate, 0.5 ml/min; wavelength, 260 nm. Calibration curves for both substances were linear in concentration ranges from 1 to 500 microg/0.1g and correlation coefficients exceeded 0.99. The relative standard deviation (R.S.D.) of the method was evaluated at concentrations of 10 and 100 microg/0.1g, and ranged from 0.84 to 10.6%. Using this method, we determined the regional distribution levels of glycerol and mannitol in various tissues from an autopsied brain dead man. PMID- 11909654 TI - Articular surfaces of the pectoral girdle: sex assessment of prehistoric New Zealand Polynesian skeletal remains. AB - Prehistoric Polynesian skeletal remains are frequently being recovered in New Zealand due to the increasing pace of urbanisation. Since such material must often be reinterred quickly, it is important that the sex of individuals be determined from the remains in a relatively short time. For this purpose, discriminant function analysis was utilised for sex determination of prehistoric adult New Zealand Polynesian clavicles (31 male and 31 female) and scapulae (33 male and 38 female). Diameters of the acromial and sternal ends of the clavicle and the height and breadth of the scapular glenoid cavity were measured and subjected to Statistical Package for the Social Sciences (SPSS) direct discriminant function analysis. For the single discriminant function derived, accuracy of sex determination was 97.7% and reduction in error over random assignment by sex was 95%. This discriminant function will be a useful tool in the assessment of human remains in the forensic and archaeological context because it incorporates measurements which can be taken on incomplete bones. PMID- 11909656 TI - Rapid analysis of 3,4-methylenedioxymethamphetamine: a comparison of nonaqueous capillary electrophoresis/fluorescence detection with GC/MS. AB - Because of the increasing use of 3,4-methylenedioxymethamphetamine (3,4-MDMA), a rapid and sensitive analytical technique is required for its detection and determination. Using nonaqueous capillary electrophoresis/fluorescence spectroscopy (NACE/FS) detection, it is possible to determine this drug at the level 0.5 ppm without any pre-treatment in less than 5 min. After liquid-liquid extraction, the sample can be condensed and a detection limit of 3,4-MDMA in urine of 50 ppb (S/N = 3) can be achieved. The precision of the method was evaluated by measuring the repeatability and intermediate precision of migration time and the corrected peak height by comparison with a 3,4-MDMA-D5 internal standard. With the conventional GC/MS method, it is necessary to derivatize the 3,4-MDMA before injection and the GC migration time also is in excess of 20 min. Therefore, NACE/FS represents a good complementary method to GC/MS for use in forensic analysis. PMID- 11909655 TI - Effects of acute cold exposure on blood-brain barrier permeability in acute and chronic hyperglycemic rats. AB - These experiments were carried out to study, the effects of cold exposure on the permeability of blood-brain barrier (BBB) in hyperglycemic rats. The integrity of the BBB was investigated using Evans blue albumin (EBA) extravasation. Serum glucose levels in hyperglycemic rats were significantly higher than that obtained from normoglycemic rats (P < 0.05). Mean arterial blood pressure in hypothermic groups significantly dropped into lower levels, than that obtained in normothermic groups (P < 0.05). The EBA extravasation to the cerebellum in the group of cold exposure+acute hyperglycemia significantly increased compared with the values obtained from the cold exposure group (P < 0.05). The EBA extravasation to the brain regions of diabetic rats exposed to cold increased more than that in normotermic control rats (P < 0.05), but did not exceed the levels in cold controls. The result of this study suggests that, acute hyperglycemia superimposed upon the permeability of BBB in the rat exposed to cold, only in selected regions of the brain, especially the cerebellum, and this result could be an important factor to explain the mechanisms of death related with hyperglycemia+cold exposure in forensic medicine. PMID- 11909657 TI - Evaluation of scientific evidence using Bayesian networks. AB - Bayesian networks provide a valuable aid for representing epistemic relationships in a body of uncertain evidence. The paper proposes some simple Bayesian networks for standard analysis of patterns of inference concerning scientific evidence, with a discussion of the rationale behind the nets, the corresponding probabilistic formulas, and the required probability assessments. PMID- 11909658 TI - On-column derivatization for determination of amphetamine and methamphetamine in human blood by gas chromatography-mass spectrometry. AB - A simple determination method of amphetamine (AP) and methamphetamine (MA) in human blood was developed using on-column derivatization and gas chromatography mass spectrometry (GC-MS). AP and MA were adsorbed on the surface of Extrelut and then derivatized the N-propoxycarbonyl derivatives using propylchloroformate. Pentadeuterated MA was used as an internal standards. The recoveries of AP and MA from the spiked blood were 89.7 and 90.3%, respectively. The calibration curves showed linearity in the range of 12.5-2000 ng/g for AP and MA in blood. The coefficients of variation of intraday and interday were 0.42-4.58%. Furthermore, this proposed method was applied to some medico-legal cases of MA intoxication. MA and its metabolite AP were detected in the blood samples, and the correlation of the blood level of amphetamines and the behaviors of the victims was in good agreement with the criteria proposed by Nagata [Jpn. J. Legal Med. 37 (1983) 513]. PMID- 11909659 TI - Light microscopical investigations on structural changes of skeletal muscle as artifacts after postmortem stimulation. AB - Samples of skeletal muscle were taken from 20 human corpses where mechanical or electrical stimulation had been carried out up to 8h postmortem (hpm) in order to estimate the time since death. The stimulation had caused an idiomuscular bulge or tetanic contraction of the muscle tissue at this location. The muscle samples were examined for structural changes of the fibers by light microscopy. A comparison with control muscle samples taken contralaterally from the same corpse, showed that the findings previously interpreted as being of intravital origin, e.g. destruction of fiber integrity, invagination and contraction bands, could also be due to postmortem alterations. It is hypothesized that structural changes to the muscle fibers can, in general, be produced as long as the manifestation time is shorter than the supravital phase after the time of force impact. PMID- 11909660 TI - "Natural death" of a patient with a deactivated implantable-cardioverter defibrillator (ICD)? AB - A 66-year-old patient with terminal heart insufficiency (NYHA IV) received maximum medical therapy, but was also in need of an implantable-cardioverter defibrillator (ICD). The ICD functioned flawlessly for the whole duration of implantation. It reverted several ventricular tachycardias with anti-tachycardial pacing alone, whereas some needed cardioversion as well. The patient died on the fourth day of hospitalization for a routine check of his ICD. The post-mortem examination revealed, that the ICD was deactivated and that the data had been erased after the patient's death. By reading off the raw data still stored within the ICD, the erased information could be restored. The stored EGMs showed traces of old ICD interventions as well as a permanent deactivation provoked by exposition to a magnetic field just hours before the patient's death. The problem of archiving and documenting the volatile electronic data inside the ICD is discussed. The need of a full autopsy after telemetric reading of the ICD data, including the explantation of the ICD aggregate and electrodes, as a means of quality assurance and under forensic aspects is emphasized. PMID- 11909661 TI - The "skin-skull-brain model": a new instrument for the study of gunshot effects. AB - In order to create and study wound morphology, a "skin-skull-brain model" had to be designed which would make the laboratory reproduction of a real ballistic injury possible. To simulate the human skin, an artificial skin (a silicon cap) is used. This silicon scalp contains synthetic fibers (artificial leather) to simulate the collagen and fat of the scalp. The artificial skull is a layered polyurethane sphere (19 cm o.d.; and 5, 6, or 7 mm thick) constructed in a specially designed form with a Tabula externa, Tabula interna, and a porous Diploe sandwiched in between. The periostium of the artificial skull is made of latex. This elastic latex layer prevents the bone fragments from scattering after the model has been struck by gunfire. The brain itself is simulated with ordnance gelatin, 10% at 4 degrees C, a material well known in wound ballistics. Gunshots were fired at a distance of 10 m from the model. During the evaluation of the "skin-skull-brain model", it was possible to show that injuries inflicted to this model are fully comparable to the morphology of equivalent real gunshot injuries. Using the "skin-skull-brain model" has some significant advantages: the model is inexpensive, easy to construct, instantly available for use, and eliminates ethics conflicts. The main advantage of such a model is, in comparison with biological substances, the high reproducibility of inflicted traumas. PMID- 11909662 TI - A study of the morphology of gunshot entrance wounds, in connection with their dynamic creation, utilizing the "skin-skull-brain model". AB - The goal of this study was to document the dynamic effects created within, and the developing mechanisms of a gunshot entrance wound to the skin utilizing high speed photography and the "skin-skull-brain model". The high-speed photography was taken with an Imacon 468/Hadland-Photonics camera. Full metal jacketed, 9 mm Luger projectiles were fired at the target model from a distance of 10 m. During the evaluation of the "skin-skull-brain model", it was possible to show that injuries inflicted to this model are fully comparable to the morphology of equivalent real gunshot entrance wounds. It has been possible to document and study the dynamic process of the "bullet-skin-interaction" in the gunshot entrance wound. The development of the morphologic terms of the entrance wound are discussed. In combination with high-speed photography, this "skin-skull-brain model" is a perfect tool for the documentation and the study of the dynamic development of gunshot entrance wounds in the skin. PMID- 11909663 TI - A "skin-skull-brain model" for the biomechanical reconstruction of blunt forces to the human head. AB - In order to create and study blunt force wound morphology, a "skin-skull-brain model" had to be designed which would make the laboratory reproduction of a blunt force injury to the head possible. During the evaluation of the "skin-skull-brain model", it was possible to show that injuries inflicted to this model are fully comparable to the morphology of equivalent real blunt forces injuries to humans. Utilization of the "skin-skull-brain model" presents some significant advantages: the model is inexpensive, easy to construct, instantly available for use, and eliminates ethics conflicts. The main advantage of such a model is, in comparison with biological substances, the high reproducibility of experimentally inflicted traumas. PMID- 11909664 TI - Demonstration of a chloroquine fatality after 10-month earth-grave. AB - A 19-year-old woman suspect of a suicidal drug intoxication was exhumed after a 10-month earth-grave, because the police was accused of manslaughter and neglected help by the relatives of the deceased. Toxicologic analysis revealed as the cause of death an acute chloroquine intoxication. An expert opinion had to deal with the question if the woman would have been saved if the police had appeared earlier. Therefore the duration of agonal period after suicidal chloroquine ingestion was important. An estimation of the time since death was possible on the one hand ex-post from the development of cadaveric changes and supravital reactions and on the other hand, based on premortal changes detectable on the body together with the findings of the authorities. Taking into account all evidence the woman was probably already dead at or prior to the arrival of the police (110 min after ingestion), at least this could not be excluded. Chloroquine has to be considered to be useful for fatal poisoning, which is also recommended in some publications on methods to commit suicide. PMID- 11909665 TI - DNA genotyping of unbuffered formalin fixed paraffin embedded tissues. AB - Formalin-induced DNA degradation was studied at different fixation times (3, 7, 16 and 32 days) each on 10 formalin fixed paraffin embedded tissues (FFPET) stored for 15 years at room temperature. The four different extraction protocols used in this study showed that Chelex100 extracts performed the best at 3 and 7 days of formalin fixation (DFF) (with regard to the quantity and the quality of the DNA). However, Qiamp extracts showed better results for long sized alleles, as well for single polymerase chain reaction (PCR) amplifications after 16 and 32 DFF, as for multiplex PCR at shorter fixation times. DNA degradation is expressed by the size of the amplified alleles, only 100 bp templates surviving after 32 DFF (AMG locus). Single locus amplifications (CD4 and FES/FPS alleles) performed better than multiplex PCR (ProfilerPlus), with nearly 100% positive results at 7 DFF. In both types of amplifications, the success rate decreased proportionally with the time of formalin fixation and, consequently, with the size of the required DNA template. PMID- 11909666 TI - Y-STR DNA amplification as biological evidence in sexually assaulted female victims with no cytological detection of spermatozoa. AB - Identification of spermatozoa is the biological evidence most often sought in specimens from rape victims. Absence of spermatozoa usually terminates biological investigations, and the victim's testimony can be contested. We assessed the utility and reliability of PCR amplification using Y-chromosomal STR polymorphisms in specimens from female victims of sexual assault with negative cytology. One hundred and four swabs without spermatozoa detected by cytology were collected from 79 alleged sexually assaulted female victims and amplification of Y-STR and of amelogenin was performed.Overall, Y-chromosome was detected and evidenced sexual penetration in 28.8% of swabs. In the population of victims examined more than 48 h after the sexual assault, Y-STR were still evidenced in 30% of the cases. These results show that swabs should be taken from victims for Y-chromosome DNA typing even after long delays between sexual assault and medical examination. PMID- 11909667 TI - Validation of the STR DXS7424 and the linkage situation on the X-chromosome. AB - X-linked microsatellite markers have proven to be powerful tools for parentage testing, mainly in deficiency paternity cases when the disputed child is female. However, only a small number of X-linked short tandem repeats (STRs) have been comprehensively described for forensic applications to date. We present sequence and population genetic data of the DXS7424 STR (GDB-G00-577-633) which is a trinucleotide repeat polymorphism representing 12 alleles of 147-180 bp in length. DXS7424 is located at Xq22 and closely linked to DXS101, corresponding to a genetic localisation of 104.9-121 cM from Xp-tel.PCR fragment length measurements and sequencing were carried out using the automatic gene analyser ABI 310 (Applied Biosystems). The population of 764 unrelated Germans checked for this STR exhibited the following features: polymorphism information content (PIC) = 0.780; heterozygosity (Het) = 0.843; mean exclusion chance (MEC = 0.766. Kinship tests revealed a typical X-linked inheritance. In 300 meioses under investigation, mutations were not found. Significant deviations from the Hardy Weinberg equilibrium (HWE) were not established. Linkage studies confirmed closely linkage to DXS101. Additional we found linkage disequilibrium between DXS7424 and DXS101. This requires to use the established haplotype frequencies in kinship testing. PMID- 11909668 TI - Factors influencing the precision of estimating the postmortem interval using the triple-exponential formulae (TEF). Part I. A study of the effect of body variables and covering of the torso on the postmortem brain, liver and rectal cooling rates in 117 forensic cases. AB - Brain, liver, rectal and environmental temperatures were continuously monitored under controlled conditions, in 117 forensic fatalities, for up to 60 h after death. Cases were studied either naked (63%) or covered with two blankets (37%). Bodies were classified into fat and thin groups. Statistical analysis and curve fitting of the data yielded the average triple-exponential formulae for each body site and each body group. The effects of covering of the torso and body parameters such as weight, height, surface area and 'cooling size factor' (Z) on postmortem cooling are assessed and discussed. Results show that covering of the torso has a significant influence on the rate of postmortem cooling, however, there is no good correlation between the body variables and the cooling rate. PMID- 11909669 TI - Factors influencing the precision of estimating the postmortem interval using the triple-exponential formulae (TEF). Part II. A study of the effect of body temperature at the moment of death on the postmortem brain, liver and rectal cooling in 117 forensic cases. AB - The temperatures of three body sites, namely, the brain, liver and the rectum as well as the temperature of the environment were continuously monitored, every 5 10 min, in 117 forensic cases commencing soon after death and in most cases, within 45 min postmortem. The body temperature at the moment of death was empirically determined by a computer-based extrapolation method. Thus, temperature data for the first 3h of each body site were fitted to single exponential equations and the fitted curve was extrapolated backwards to obtain the intercept on the Y-axis (the temperature axis). The effect of body temperature at the moment of death on postmortem cooling rate is examined and factors influencing body temperature at death are discussed. Forensic fatalities associated with hyper and hypothermia are reviewed briefly. PMID- 11909670 TI - A study of the shape of the post-mortem cooling curve in 117 forensic cases. AB - The shape of the post-mortem cooling curve is described and its theoretical aspects are summarised. The concepts of the "infinite cylinder" and the "initial temperature plateau" are explained and their practical implications are discussed. Results of the study of the shape of post-mortem cooling curves for the brain, liver and the rectum of 117 cases are given. The bodies were monitored either naked or covered with blankets. For each case, temperatures of the three body sites and the environment were monitored soon after death and up to many hours post-mortem. Empirically derived three-exponential formulae were used in this study. The cooling curves for the three body sites were found to be of compound shapes and the slopes of the curves vary throughout the monitoring period. The "initial temperature plateau" was found on average in 22% of all cooling curves with the plateau incidence being significantly highest in the rectal curves (27% of rectal curves compared with 7% of brain and liver curves, P<0.1%). The effect of body sites, body build and covering of the torso on the occurrence of the plateau is assessed and discussed. PMID- 11909671 TI - Computer aided shot reconstructions by means of individualized animated three dimensional victim models. AB - In cases of lethal firearm injuries with indefinite indications concerning self versus third-party infliction a computer enhanced reconstruction with the aim of an anatomical feasibility study can provide significant clues concerning the course of the traumatic event. To this end an exact three-dimensional geometrical model of the victim including all relevant anatomical data as well as the careful documentation of the injuries and a three-dimensional model of the characteristic outlines of the weapon true to scale is generated with the help of an animation program (POSER Version 4, Meta Creation, Egisys AG). With this animated digital three-dimensional model of the victim and the weapon a series of simulation sequences is created by variation of the body positions and the grasp of the weapon. Anatomically impossible positions in view of the physical characteristics of the victim and the site and direction of the bullet path are automatically excluded from the reconstruction. An exact match of the simulation sequence and the real injuries is a statement for a possible self-infliction of the gunshot wound. PMID- 11909672 TI - An unusual airplane crash--deadly life saver. Unintentional activation of an automated reserve opening device causing airplane accident. AB - A Pilatus PC 6 "Turbo-Porter" crashed after having dropped skydivers. The interdisciplinary investigation of the crash revealed that the automated reserve opening device "Student-CYPRES" which the passenger wore on his parachute system had been set off during the landing approach. The parachute pulled the passenger out of the airplane. Subsequently the parachute got caught at the horizontal tail, and its wearer was instantly killed by internal decapitation. The airplane crashed because of its damaged horizontal tail, and the pilot lost his life. PMID- 11909673 TI - Effects of refrigeration on the biometry and development of Protophormia terraenovae (Robineau-Desvoidy) (Diptera: Calliphoridae) and its consequences in estimating post-mortem interval in forensic investigations. AB - The aim of this study was to simulate the low temperatures that insects could experience between the time being sampled from cadavers and their arrival in the laboratory. This was in order to investigate the effect of low temperature on development of maggots. At different stages of development, individuals of Protophormia terraenovae (Robineau-Desvoidy) reared at 24 degrees C were submitted to a temperature of 4.0+/-0.5 degrees C for a period varying from 1 to 10 days. Independent of the stage of development at which the insects were refrigerated, the treatment induced significant changes on the duration of development. The effect of low temperature on the developmental time between the return to 24 degrees C to adult emergence depended on the larval stage that was refrigerated. When first instar larvae and prepupae were refrigerated, the time to emergence at 24 degrees C decreased with an increase of duration of the refrigeration period. Time to emergence increased under the same conditions when second instar larvae and pupae were refrigerated. These results indicate that keeping larvae of P. terraenovae at 4 degrees C does not just simply lead to a cessation of metabolism but disturbs the regular development. Ten days of cooling induced an error in estimating post-mortem interval (PMI) of more than 6h. PMID- 11909675 TI - Retrograde transthoracic venous bullet embolism. Report of a case following a single gunshot with multiple wounds in the left arm and chest. AB - Bullet embolism of the arterial, venous and paradoxical types are well known but rare complications of penetrating missile injuries. Retrograde transthoracic venous bullet embolization is extremely rare with only four cases previously reported in the literature. Single gunshot can cause multiple wounds, mainly because the bullet passes through an intermediate target before striking the victim. We present the autopsy findings of a retrograde transthoracic bullet embolism to the right external iliac vein after a single gunshot with multiple wounds in the left arm and thorax. Problems related to medico-legal investigations of gunshots are reviewed. PMID- 11909674 TI - Evaluation of diagnostic tools applied in the examination of sudden unexpected deaths in infancy and early childhood. AB - During the period between 1984 and 1999, 309 cases of sudden unexpected death in infancy and early childhood (0-3 years) were investigated at the Institute of Forensic Medicine in Oslo. In 73 cases, an explainable cause of death was found. In this non-sudden infant death syndrome (SIDS) group, 42 cases were due to disease, 14 to accidents, 7 to neglect/abuse and 10 cases were due to homicide. In 43 cases, there were pathological findings at the autopsy or suspect features in the history and/or circumstances, which were, however, insufficient to explain death ("borderline" SIDS). In the remaining 193 cases, nothing of significance was detected ("pure" SIDS). The purpose of the present study was to evaluate the importance of the different diagnostic tools used in diagnosing non-SIDS and borderline SIDS cases. The definition of SIDS requires a negative history as well as a negative autopsy result. Thus, the following variables were analysed: circumstances, medical history and autopsy, which included a gross pathological investigation, histology, neuropathology, microbiology, radiology and toxicology. In diagnosing deaths due to disease, histology, neuropathology and microbiology were the most important diagnostic tools. In contrast, information about the circumstances of death and the gross pathological findings at autopsy most often revealed the cause of death in accidents and cases of neglect/abuse and homicide. Following the drop in SIDS rate in Norway after 1989, the share of pure SIDS in proportion to the total population of sudden unexpected deaths in infancy and early childhood has decreased. The increasing proportion of non-SIDS and borderline SIDS cases presents a challenge to improve the quality of the investigation in cases of sudden death in infancy and early childhood. PMID- 11909676 TI - Y-STR haplotype data and allele frequency of the DXS10011 locus in a Japanese population sample. AB - The distribution of allele frequency of X-chromosomal STR, DXS10011, from 99 unrelated Japanese, 72 male and 27 female, were determined by PCR amplification and PAGE. At the same time, haplotype frequencies of five Y-chromosomal STR loci, DYS19, DYS389I, DYS389II, DYS390 and DYS393 from male samples were determined. PMID- 11909677 TI - Motherless case in paternity testing by Lee et al. PMID- 11909678 TI - Leiden research program on ageing. AB - Research into ageing is among the priorities of the Leiden University Medical Center. Several tight collaborations between basic and clinical departments are the basis of this program. Our focus is to identify determinants of human longevity and disease at old age with an emphasis on inflammation, atherosclerosis, and cognitive decline. To this end we enroll a large series of long-lived families for genetic screening, prospectively follow large cohorts of old people dependent on various genetic and environmental risk factors, and perform randomized controlled trials in the general population testing plausible hypotheses how interventions can maximize rewarding lifespans. PMID- 11909679 TI - The reserve-capacity hypothesis: evolutionary origins and modern implications of the trade-off between tumor-suppression and tissue-repair. AB - Antagonistic pleiotropy, the evolutionary theory of senescence, posits that age related somatic decline is the inevitable late-life by-product of adaptations that increase fitness in early life. That concept, coupled with recent findings in oncology and gerontology, provides the foundation for an integrative theory of vertebrate senescence that reconciles aspects of the 'accumulated damage' 'metabolic rate', and 'oxidative stress' models. We hypothesize that (1) in vertebrates, a telomeric fail-safe inhibits tumor formation by limiting cellular proliferation. (2) The same system results in the progressive degradation of tissue function with age. (3) These patterns are manifestations of an evolved antagonistic pleiotropy in which extrinsic causes of mortality favor a species optimal balance between tumor suppression and tissue repair. (4) With that trade off as a fundamental constraint, selection adjusts telomere lengths--longer telomeres increasing the capacity for repair, shorter telomeres increasing tumor resistance. (5) In environments where extrinsically induced mortality is frequent, selection against senescence is comparatively weak as few individuals live long enough to suffer a substantial phenotypic decline. The weaker the selection against senescence, the further the optimal balance point moves toward shorter telomeres and increased tumor suppression. The stronger the selection against senescence, the farther the optimal balance point moves toward longer telomeres, increasing the capacity for tissue repair, slowing senescence and elevating tumor risks. (6) In iteroparous organisms selection tends to co ordinate rates of senescence between tissues, such that no one organ generally limits life-span. A subsidiary hypothesis argues that senescent decline is the combined effect of (1) uncompensated cellular attrition and (2) increasing histological entropy. Entropy increases due to a loss of the intra-tissue positional information that normally regulates cell fate and function. Informational loss is subject to positive feedback, producing the ever accelerating pattern of senescence characteristic of iteroparous vertebrates. Though telomere erosion begins early in development, the onset of senescence should, on average, be deferred to the species-typical age of first reproduction, the balance point at which selection on this trade-off should allow exhaustion of replicative capacity to overtake some cell lines. We observe that captive-rodent breeding protocols, designed to increase reproductive output, simultaneously exert strong selection against reproductive senescence and virtually eliminate selection that would otherwise favor tumor suppression. This appears to have greatly elongated the telomeres of laboratory mice. With their telomeric failsafe effectively disabled, these animals are unreliable models of normal senescence and tumor formation. Safety tests employing these animals likely overestimate cancer risks and underestimate tissue damage and consequent accelerated senescence. PMID- 11909680 TI - Extension of life span and stress resistance of Drosophila melanogaster by long term supplementation with melatonin. AB - According to the free radical theory of aging, free radicals are involved in the production of changes in cellular metabolism that lead to a time-dependent functional decline in all living beings. Consequently, antioxidant and/or free radicals scavengers may retard the aging process. We explored the effect of melatonin on the life span of Drosophila melanogaster (Oregon wild strain). It was presumed that given the antioxidant and free radicals scavenger properties of melatonin, this hormone would prevent oxidative damage to the fly tissues and slow down the process of aging. Melatonin, added daily to the nutrition medium at a concentration of 100 microg/ml, increased significantly the life span of D. melanogaster. The maximum life span was 61.2 days in controls and 81.5 days in melatonin fed flies. Relative to the controls, the percentage increase in the melatonin fed flies was 33.2% in maximum life span, 19.3% in the onset of 90% mortality, and 13.5% in median life span. Furthermore, in a test of superoxide mediated toxicity it was shown that melatonin treatment increased the resistance of D. melanogaster to paraquat. Finally, the augmented resistance to an ambient temperature of 36 degrees C was also a demonstration of the antioxidative protection provided by the hormone. PMID- 11909681 TI - Oxidative phosphorylation enzyme complexes in caloric restriction. AB - Free radicals, generated especially by electron leakage from mitochondrial electron transport chain (ETC), are accepted as one of the possible causes of aging. Long-term caloric restriction (CR) is known to increase the species specific average and maximum life spans. Thus it provides a means for investigating mechanisms of aging. There is evidence suggesting a decrease in the free radical production with CR. In this study, Blue-Native PAGE (BN-PAGE) technique was used to investigate the effect of CR on the oxidative phosphorylation enzyme complexes. Of the total 30 female Swiss Albino balb/c mice, 15 were used as control and the other 15 as CR group. Alternate day feeding regimen was used in the CR group for 66 weeks beginning at the end of 3rd month. In the control group, 5 (33.3%) mice died, 3 (20%) of them of breast cancer, 2 (13.3%) of unknown causes and no death cases were observed in the CR group during the study. BN-PAGE was performed on the extracts from brain mitochondrial fractions. Complexes II and V were excluded from the study due to some analytical limitations. No difference was found in the levels of complexes I and III between the groups. In the CR group, complex IV level was found increased and the ratio of complex III-IV decreased compared with the control group. Since there is a slight increase (108%) in the level of complex IV in the CR group, our results could suggest possible partial compensation of electron leakage in the upstream complexes in ETC, and the decrease of free radical production with CR. PMID- 11909683 TI - Reduced susceptibility to peroxidation of erythrocyte plasma membranes from centenarians. AB - The plasma membrane composition affects intracellular processes and the cellular susceptibility to free radical attack, which has been associated with the impairment of cellular functions occurring during senescence. The study of the modifications of the plasma membrane in centenarians might elucidate the biological mechanisms at the basis of longevity and successful aging. The work was performed in 190 subjects, divided into five groups according to the age range: (1) 21-40 years (n=25); (2) 41-60 years (n=30); (3) 61-80 years (n=30); (4) 81-99 years (n=50); and (5) centenarians (> or = 100 years) (n=55). The following determinations were performed on erythrocyte membranes: (i) the lipid peroxide level (Lp) evaluated as malondialdehyde content; (ii) susceptibility to in vitro oxidation evaluated as difference in the content of thiobarbituric acid reactive substances before and after phenylhydrazine addition; (iii) unsaturated/saturated fatty acid ratio and individual polyunsaturated fatty acid composition measured by gas chromatography; and (iv) fluidity studied by means of the anisotropy of the probe 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). Erythrocyte membranes from centenarians showed: (i) decreased basal lipid peroxide levels and reduced susceptibility to peroxidation in comparison with elderly subjects; (ii) increased unsaturated/saturated fatty acid ratio in comparison with every other age group; (iii) higher levels of eicosapentaenoic and docosahexaenoic acid and reduced content of linoleic and arachidonic acid in comparison with elderly subjects; and (iv) decreased anisotropy of TMA-DPH, i.e. higher fluidity compared with all the other age groups. In conclusion, the present work demonstrates that erythrocyte membranes from centenarians show some distinct features in comparison with elderly subjects that might act in a protective way against injuries. PMID- 11909682 TI - Oxidative DNA damage as a marker of aging in WI-38 human fibroblasts. AB - Cause-effect relationships between oxidative stress, DNA damage and aging were investigated in WI-38 human diploid fibroblasts at 21, 41 or 58 population doublings (PDs), corresponding to young, middle age or old fibroblasts, respectively. Oxidative DNA damage was evaluated by immunohistochemical detection of 8-hydroxy-2'deoxyguanosine (8-OHdG) adducts or by single cell microgel electrophoresis (COMET assay). Aging was evaluated by growth rate, senescence associated-beta-galactosidase (SA-beta galactosidase) activity, cell cycle distribution, and expression of p21. Our results demonstrate that (i) oxidative DNA damage is proportional to the age of cells (ii) DNA damage in old/58 PDs cells reflects both an increased susceptibility to oxidative stress, induced by acute exposure to sub-lethal concentrations of hydrogen peroxide (H(2)O(2)), and a reduced efficiency of repair mechanisms. We also show that mild chronic oxidative stress, induced by prolonged exposure to 5 microM H(2)O(2), accelerates aging in fibroblasts. In fact, this treatment increased 8-OHdG levels, SA-beta galactosidase activity, and G0/G1 cell cycle arrest in middle age/41 PDs, making them similar to H(2)O(2)-untreated old/58 PDs cells. Although other mechanisms may concur in mediating the effects of H(2)O(2), these results lend support to the concept that oxidative stress may be a key determinant of aging. Measurements of oxidative DNA damage might therefore be exploited as reliable marker of cellular aging. PMID- 11909684 TI - Attenuation of Ca(2+)-activated ATPase and shortening velocity in hypertrophied fast twitch skeletal muscle from aged Japanese quail. AB - The effect of aging on the in vitro contractile properties of the patagialis (PAT) muscle of 35 young adult (YA; 8 weeks of age) and 35 aged adult (AA, 110 weeks of age) Coturnix quails was examined after 0-30 days of stretch-overload. Overload was achieved by placing a weight equivalent to 12% of the birds' body weight on one wing. The contralateral wing served as the intra-animal control. Overload increased the weight of the PAT by 45.1+/-2.1% in YA, and 24.1+/-2.6% in AA. Twitch contraction time increased with loading from 43.2+/-1.2 to 67.3+/-2.2 ms in YA birds and 57.2+/-1.7 to 77.4+/-1.9 ms in AA birds. Unloaded shortening velocity (Vo) decreased by 40.1+/-2.2 and 38.8+/-3.2% in YA and aged birds, respectively. The decrease in fast myosin expression was greater in overloaded muscles of YA (20%) as compared to AA birds (12%). However, this was accompanied by a greater decrease in total muscle ATPase activity in aged birds (61%) compared to YA birds (40%). Myosin isozyme Ca(2+)-ATPase activity was 26% lower in FM1 but not other fast myosins in YA birds, but it was approximately 30% lower in all fast myosins in PAT muscles of aged birds. These data show that the reduction of Vo and the increase in twitch duration with aging may be due in part to reductions in ATPase activity in all myosin isoforms, as compared to myosin isoforms isolated from YA birds. PMID- 11909685 TI - Troglitazone treatment of aging Brown Norway rats improves food intake and weight gain after fasting without increasing hypothalamic NPY gene expression. AB - Compared to younger animals, aged male Brown Norway (BN) rats demonstrate increased body fat and serum insulin, and lower prepro-neuropeptide Y (ppNPY) mRNA content in the arcuate nucleus (ARC), and blunted food intake (FI) and body weight (BW) gain in response to a 72 h fast. Since centrally administered insulin decreases FI and weight of young rats and inhibits fasting-induced increases of NPY gene expression, we hypothesized that hyperinsulinemia in old rats contributes to an age-related central dysregulation of energy balance. Young, middle-aged and old BN rats were fed chow with troglitazone (Trog; 200 mg/kg BW/d) or without drug for 75 d (Experiment 1) or 66 d (Experiment 2). Rats were then fasted for 72 h, refed for 2 weeks and sacrificed after an overnight fast (Experiment 1) or fasted for 72 h and sacrificed (Experiment 2). Serum insulin and leptin were measured from trunk blood and brains were analyzed for ppNPY mRNA by in situ hybridization. In Experiment 1, troglitazone treatment resulted in increased post-fast weight gain, rate of gain and FI in old rats. Troglitazone decreased serum insulin by 50% in old rats, while leptin levels decreased 20-30% in all age groups in Experiment 1. No differences in serum insulin or leptin were detectable with troglitazone treatment in Experiment 2, due to the extreme suppression caused by the 72 h fast. Troglitazone treatment did not increase ARC NPY gene expression either after a 72 h fast and re-feeding for 2 weeks (Experiment 1) or immediately after a 72 h fast (Experiment 2). These findings suggest that increased insulin levels may contribute to age-related impairments of FI and BW regulation. However, improvements in these defects in energy regulation induced by troglitazone do not appear to result from changes in NPY gene expression, and may be due to alterations in other hypothalamic neuropeptides that regulate energy balance. PMID- 11909686 TI - Asymptomatic bacteriuria in elderly humans is associated with increased levels of circulating TNF receptors and elevated numbers of neutrophils. AB - Low-grade inflammatory activity is strongly associated with age-associated diseases such as atherosclerosis, dementia, type-2 diabetes, sarcopenia, and osteoporosis and predicts mortality risk in elderly populations. The aim of the current study was to investigate if asymptomatic bacteriuria in elderly humans was associated with inflammation. Midstream clean-catch urine culture was collected from consecutive, elderly patients at admission to a department of internal medicine due to functional disability. Forty patients (age 70-91 years) were selected and included in the current study; 20 subjects had positive urine culture and 20 sex- and age-matched subjects had negative urine culture. Inclusion criteria were temperature below 37.8 degrees C, no clinical signs of infection and no current antibiotic treatment. Patients with asymptomatic bacteriuria had significantly increased levels of circulating tumor necrosis factor receptors (sTNFR-I) and a higher number of neutrophils in the blood compared to the group without bacteriuria. Thus, the present study provides some support for the hypothesis that asymptomatic urinary infections are associated with low-grade immune activity in frail, elderly humans. PMID- 11909687 TI - Dehydroepiandrosterone-sulfate serum levels and common age-related diseases: results from a cross-sectional Italian study of a general elderly population. AB - The association of low serum dehydroepiandrosterone sulfate (DHEAS) levels with age, lifestyle, general health status indicators, and specific diseases was investigated in 436 men and 544 women of 65-97 yr old. In both sexes low serum DHEAS levels were associated with age, alcohol intake, number of current medications, and decreased thyroid function. Low DHEAS was also associated with low serum albumin in men and low systolic blood pressure in women. Compared to healthy men (n=106) age-adjusted serum DHEAS levels were significantly lower in men with atrial fibrillation, chronic obstructive lung disease, dementia, parkinsonism, cancer, diabetes, hypothyroidism, and in institutionalized men. Compared to healthy women (n=100) age-adjusted serum DHEAS levels were significantly lower in women with occlusive arterial disease, chronic obstructive lung disease, and osteoporosis. After controlling for differences in lifestyle and general health status parameters, low DHEAS levels remained statistically associated only with atrial fibrillation in men and osteoporosis in women, and it cannot be excluded that these association were spurious, due to multiple comparisons. These data suggest that in elderly people low serum DHEAS levels are more a non-specific indicator of aging and health status than a risk indicator of specific diseases. PMID- 11909689 TI - Proteomics in experimental gerontology. AB - The first gerontological studies using two-dimensional gel electrophoresis (2DGE) were frustrating since it was very difficult, when not impossible, to identify the proteins for which an age-related change in expression level was suspected. Reproducibility was also a main pitfall. Accumulated progress in 2DGE and especially the development of mass spectrometry of proteins and peptides gave accessibility to the routine identification of differentially expressed proteins. A new paradigm was born: proteomics. In addition to expression changes, post translational modifications are included in proteomics, and will be more and more studied using mass spectrometry. After a review of the current developments of 2DGE and mass spectrometry, we shall discuss how the technologies currently available in proteomics could give fresh impetus to experimental gerontology, complementary to more recent approaches based on wide expression analysis tools such as DNA and protein arrays. PMID- 11909688 TI - Correlation of age with in vivo expression of endothelial markers. AB - The importance of endothelial senescence as a pathogenetic factor in age-related vascular alterations has almost exclusively been studied in vitro. However, the in vitro-findings have rarely been compared with histomorphological changes in aging human tissue or in age-related degenerative diseases. Therefore, we compared the expression of the endothelial marker CD34 and the procoagulant protein von Willebrand factor (vWf) in lung endothelium by conventional immunohistochemistry and confocal laser scan microscopy (CLSM), taking the age of the patients into consideration. The staining reactions were statistically analysed by covariance analysis. With age the endothelial staining intensity of CD34 increased in arteries and veins, but decreased in arterioles, capillaries and venules. For vWf, on the contrary, the endothelial staining intensity increased with age in all types of vessels. CLSM confirmed a mosaic staining pattern. This study demonstrates age-associated phenotypical alterations of CD34 and vWf. Whether the down-regulation of CD34 correlates with an age-associated reduction of the angiogenetic properties of EC or an age-related over-expression of vWf as a relevant cofactor for the raised coagulatory activity and the increase in thrombotic diseases resp coronary heart disease in older patients, remains subject to debate. PMID- 11909690 TI - Introduction-serial review: iron and cellular redox status. PMID- 11909691 TI - Iron release, oxidative stress and erythrocyte ageing. AB - Iron, to be redox cycling active, has to be released from its macromolecular complexes (ferritin, transferrin, hemoproteins, etc.). Iron is released from hemoglobin or its derivatives in a nonprotein-bound, desferrioxamine-chelatable form (DCI) in a number of conditions in which the erythrocytes are subjected to oxidative stress. Such conditions can be related to toxicological events (haemolytic drugs) or to physiological situations (erythrocyte ageing, reproduced in a model of prolonged aerobic incubation), but can also result from more subtle circumstances in which a state of ischemia-reperfusion is imposed on erythrocytes (e.g., childbirth). The released iron could play a central role in oxidation of membrane proteins and senescent cell antigen (SCA) formation, one of the major pathways for erythrocyte removal. Iron chelators able to enter cells (such as ferrozine, quercetin, and fluor-benzoil-pyridoxal hydrazone) prevent both membrane protein oxidation and SCA formation. The increased release of iron observed in beta-thalassemia patients and newborns (particularly premature babies) suggests that fetal hemoglobin is more prone to release iron than adult hemoglobin. In newborns the release of iron in erythrocytes is correlated with plasma nonprotein-bound iron and may contribute to its appearance. PMID- 11909692 TI - Deleterious iron-mediated oxidation of biomolecules. AB - Iron is an essential metal for most biological organisms. However, if not tightly controlled, iron can mediate the deleterious oxidation of biomolecules. This review focuses on the current understanding of the role of iron in the deleterious oxidation of various biomolecules, including DNA, protein, lipid, and small molecules, e.g., ascorbate and biogenic amines. The effect of chelation on the reactivity of iron is also addressed, in addition to iron-associated toxicities. The roles of the iron storage protein ferritin as both a source of iron for iron-mediated oxidations and as a mechanism to safely store iron in cells is also addressed. PMID- 11909693 TI - Extracellular cysteines define ectopeptidase (APN, CD13) expression and function. AB - Alanyl aminopeptidase (APN) is a surface-bound metallopeptidase that processes the N-terminals of biologically active peptides such as enkephalins, angiotensins, neurokinins, and cytokines. It exerts profound activity on vital processes such as immune response, cellular growth, and blood pressure control. Inhibition of either APN gene expression or its enzymatic activity severely affects leukocyte growth and function. We show here that oxidoreductase-mediated modulations of the cell surface thiol status affect the enzymatic activity of APN. Additional evidence for the pivotal role of extracellular cysteines in the APN molecule was obtained when substitution of any of these six cysteines caused complete loss of surface expression and enzymatic activity. In contrast, the transmembrane Cys24 appears to have no similar function. Enzymatically inactive cysteine mutants were retained in the endoplasmic reticulum as shown by high resolution imaging and Endoglycosidase H digestion. In the absence of any crystal structure data, the demonstration that individual extracellular cysteines contribute to APN expression and function appears to be of particular importance. The data are the first to show thiol-dependent modulation of the activity of a typical surface-bound peptidase at the cell surface, probably reflecting a general regulating mechanism. This may relate to various disease processes such as inflammation or malignant transformation. PMID- 11909694 TI - Flavonoids of Inula britannica protect cultured cortical cells from necrotic cell death induced by glutamate. AB - We previously reported 12 antioxidative flavonoids isolated from the n-BuOH extract of Inula britannica (Asteraceae). This prompted us to investigate further whether these flavonoids also showed antioxidative activity upon live cells grown in a culture system. Among the 12 flavonoids tested, only patuletin, nepetin, and axillarin protected primary cultures of rat cortical cells from oxidative stress induced by glutamate. These flavonoids exerted significant neuroprotective activity when they were administered either before or after the glutamate insult. Treatment with these flavonoids maintained the activities of such antioxidant enzymes as catalase, glutathione-peroxidase, and glutathione reductase, all of which play important roles in the antioxidative defense mechanism. Moreover, these three flavonoids also attenuated significant drops in glutathione induced by glutamate which is a routine concomitant of oxidative stress by inhibiting glutathione diminution. Accordingly, these flavonoids did not stimulate the synthesis of glutathione. With regard to structure-activity relationships, our results indicated that the 6-methoxyl group in the A ring and the 3', 4'-hydroxyl groups in the B ring are crucial for the protection against the oxidative stress; glycosylation greatly reduced their protective activities. Collectively, these results indicated that patuletin, nepetin, and axillarin strongly protect primary cultured neurons against glutamate-induced oxidative stress. PMID- 11909695 TI - Superoxide organic chemistry within the liposomal bilayer, part II: a correlation between location and chemistry. AB - Coumarin ester derivatives 1, substituted at C-4 and/or C-12 with alkyl chains, were synthesized and intercalated within DMPC liposomal bilayers. By correlating the 13C chemical shift with medium polarity [E(T)(30)], the relative location of these substrates within the liposomal bilayer was determined. The length of the alkyl chain substituents clearly influences the lipophilicity of the substrates and their location and orientation within the liposome: Superoxide readily saponifies the C-12 esteric linkage of 1, when this reaction site lies in a polar region of the liposome (E(T)(30) > 45 kcal/mol), to give the corresponding 7 hydroxy coumarin derivatives 2. However, when C-12 lies deeper and is hence less available to O(2)(*-), the lactonic carbon C-2, which lies in a shallower region (E(T)(30) = 43-49), is the preferred site for superoxide-mediated cleavage. When coumarin 1 is disubstituted with long chains at both C-12 and C-4, these derivatives lie deep within the bilayer and react only slowly with O(2)(*-). These results indicate there is indeed a correlation between location within the bilayer and substrate reactivity. Contrary to the suggestion of Dix and Aikens (Chem. Res. Toxicol.6:2-18; 1993) superoxide can penetrate deep within the liposomal bilayer. Nevertheless, its concentration drops precipitously (to approximately 16% of what it is near the interface) below E(T) values of 38, thereby precluding substantial reaction with many highly lipophilic substrates. This work also confirms the findings of others that reactions of small oxy radicals occur within cellular membranes and appear to be of significant biological importance. PMID- 11909696 TI - Humic acid induces the generation of nitric oxide in human umbilical vein endothelial cells: stimulation of nitric oxide synthase during cell injury. AB - Humic acid (HA) has been implicated as an etiological factor in the peripheral vasculopathy of blackfoot disease (BFD). In this study, we examined the effects of HA upon the generation of nitric oxide (NO) during the process of lethal cell injury in cultured human umbilical vein endothelial cells (HUVECs). NO production was measured by the formation of nitrite (NO(2)(-)), the stable end-metabolite of NO. Cell death was assessed by measuring the release of intracellular lactate dehydrogenase (LDH). Treatment HUVECs with HA at a concentration of 50, 100, and 200 microg/ml concentration-dependently increased nitrite levels, reaching a peak at 12 h subsequent to HA treatment, with a maximal response of approximately 400 pmole nitrite (from 1 x 10(4) cells). HA-induced nitrite formation was blocked completely by N(G)-nitro-L-arginine methyl ester (L-NAME) and also by N(G)-methyl L-arginine (L-NMA), both being specific inhibitors of NO synthase. The LDH released from endothelial cells was evoked at from 24 h after the addition of HA (50, 100, 200 microg/ml) in a concentration- and time-dependent manner. The HA induced LDH release was also reduced by the presence of both L-NAME and L-NMA. The addition of Ca(2+) chelator (BAPTA) inhibited both nitrite formation and LDH release by HA. Moreover, the antioxidants (superoxide dismutase, vitamin C, vitamin E) and protein kinase inhibitor (H7) effectively suppressed HA-induced nitrite formation. These results suggest that HA treatment of endothelial cells stimulates NO production, which can elicit cell injury via the stimulation of Ca(2+)-dependent NO synthase activity by increasing cytosolic Ca(2+) levels. Because the destruction of endothelial cells has been implicated in triggering the onset of BFD, the induction of excessive levels of NO and consequent endothelial-cell injury may be important to the etiology of HA-induced vascular disorders associated with BFD for humans. PMID- 11909697 TI - Immunohistochemical localization of the antioxidant enzymes biliverdin reductase and heme oxygenase-2 in human and pig gastric fundus. AB - The intrinsic antioxidant capacities of the bile pigments biliverdin and bilirubin are increasingly recognized since both heme degradation products can exert beneficial cytoprotective effects due to their scavenging of oxygen free radicals and interaction with antioxidant vitamins. Several studies have been published on the localization of the carbon monoxide producing enzyme heme oxygenase-2 (HO-2), which concomitantly generates biliverdin; histochemical data on the distribution of biliverdin reductase (BVR), converting biliverdin to bilirubin, are still very scarce in large mammals including humans. The present study revealed by means of immunohistochemistry the presence of BVR and HO-2 in mucosal epithelial cells and in the endothelium of intramural vessels of both human and porcine gastric fundus. In addition, co-labeling with the specific neural marker protein-gene product 9.5 (PGP 9.5) demonstrated that both BVR and HO-2 were present in all intrinsic nerve cell bodies of both submucous and myenteric plexuses, while double labeling with c-Kit antibody confirmed their presence in intramuscular interstitial cells of Cajal (ICC). Our results substantiate the hypothesis that BVR, through the production of the potent antioxidant bilirubin, might be an essential component of normal physiologic gastrointestinal defense in man and pig. PMID- 11909698 TI - Nitric oxide augments voltage-gated P/Q-type Ca(2+) channels constituting a putative positive feedback loop. AB - P/Q-type Ca(2+) channels, which are postulated to play major roles in synaptic transmission, are regulated in a variety of ways. Ca(2+) currents through P/Q type Ca(2+) channels (Ca(v)2.1/beta(1a)/alpha(2)delta) heterologously expressed in mammalian cells were recorded using the whole-cell patch clamp method. The oxidant H(2)O(2) increased the current amplitude and the effect was reversed by the reducing agent dithiothreitol (DTT). The stimulatory effect of H(2)O(2) on the Ca(2+) current was mimicked by the NO donors, SNAP, and diethylamine NONOate, and reversed by the reducing agent DTT. The presence of a soluble guanylate cyclase inhibitor did not abolish the ability of SNAP to increase the Ca(2+) current. Adenovirus-mediated overexpression of nitric oxide synthase in combination with application of the Ca(2+) ionophore A23187 also increased the Ca(2+) current amplitude and the effect was again reversed by DTT. The NOS inhibitor L-NAME abolished the stimulatory effect of A23187, and A23187 did not change the Ca(2+) currents in the cells treated with control adenovirus particles. The time course of the decline of the Ca(2+) current, but not of the Ba(2+) current, in response to repeated depolarization was markedly slowed by adenovirus-mediated overexpression of nitric oxide synthase. The results demonstrate that nitric oxide enhances the channel activity by promoting oxidation and suggest that Ca(2+), nitric oxide synthase, and nitric oxide could constitute a positive feedback loop for regulation of voltage-gated P/Q-type Ca(2+) channels. PMID- 11909700 TI - Moderate G6PD deficiency increases mutation rates in the brain of mice. AB - Mice that harbored the x-ray-induced low efficiency allele of the major X-linked isozyme of glucose-6-phospate dehydrogenase (G6PD), Gpdx(a-m2Neu), and, in addition, harbored the transgenic shuttle vector for the determination of mutagenesis in vivo, pUR288, were employed to further our understanding of the interdependence of general metabolism, oxidative stress control, and somatic mutagenesis. The Gpdx(a-m2Neu) mutation conferred moderate G6PD deficiency in hemizygous males (Gpdx(a-m2Neu/y)) displaying residual enzyme activities of 27% in red blood cells and 13% in brain (compared to wild-type controls, Gpdx(a/y) males). In spite of this mild phenotype, the brains of G6PD-deficient males exhibited a significant distortion of redox control ( approximately 3-fold decrease in the ratio of reduced glutathione to oxidized glutathione), a considerable accumulation of promutagenic etheno DNA adducts ( approximately 13 fold increase in ethenodeoxyadenosine and approximately 5-fold increase in ethenodeoxycytidine), and a substantial elevation of somatic mutation rates ( approximately 3-fold increase in mutant frequencies in lacZ, the target and reporter gene of mutagenesis in the shuttle vector, pUR288). The mutation pattern in the brain was dominated by illegitimate genetic recombinations, a presumed hallmark of oxidative mutagenesis. These findings suggested a critical function for G6PD in limiting oxidative mutagenesis in the mouse brain. PMID- 11909699 TI - Redox-sensitive interaction between KIAA0132 and Nrf2 mediates indomethacin induced expression of gamma-glutamylcysteine synthetase. AB - Exposure of HepG2 cells to nonsteroidal anti-inflammatory drugs (i.e., indomethacin and ibuprofen; NSAIDs) as well as resveratrol, caused increased expression of the mRNAs coding for the catalytic (Gclc) and modifier (Gclm) subunits of the glutathione synthetic enzyme, gamma-glutamylcysteine synthetase. In addition, indomethacin exposure increased intracellular glutathione content as well as inhibited glutathione depletion and cytotoxicity caused by diethyl maleate. Indomethacin-induced increases in the expression of gamma glutamylcysteine synthetase mRNA were preceded by increases in steady state levels of intracellular pro-oxidants and glutathione disulfide accumulation. Simultaneous incubation with the thiol antioxidant N-acetylcysteine (NAC) inhibited indomethacin-mediated increases in GCLC mRNA, suggesting that increases in GCLC message were triggered by changes in intracellular oxidation/reduction (redox) reactions. Indirect immunofluorescence using intact cells demonstrated that indomethacin induced the nuclear translocation of Nrf2, a transcription factor believed to regulate GCLC expression. Immunoprecipitation studies showed that indomethacin treatment also inhibited Nrf2 tethering to KIAA0132 (the human homolog of Keap1 accession #D50922), which is believed to be a negative regulator of Nrf2. Consistent with this idea, over-expression of Nrf2 increased GCLC reporter gene expression and over-expression of KIAA0132 inhibited GCLC reporter gene activity as well as inhibited indomethacin-induced increases in the expression of GCLC. Finally, simultaneous treatment with NAC inhibited both indomethacin-induced release of Nrf2 from KIAA0132 and indomethacin-induced nuclear translocation of Nrf2. These results demonstrate that NSAIDs and resveratrol cause increases in the expression of gamma-glutamylcysteine synthetase mRNA and identify these agents as being capable of stimulating glutathione metabolism. These results also support the hypothesis that indomethacin-induced transcriptional activation of GCLC involves the redox dependent release of KIAA0132 from Nrf2 followed by the nuclear translocation of Nrf2. PMID- 11909701 TI - Thiophene derivatives: a new series of potent norepinephrine and serotonin reuptake inhibitors. AB - A series of (1S,3S,6R,10S)-(Z)-9-(thienylmethylene- or substituted thienylmethylene)-7-azatricyclo[4.3.1.0(3,7)]decanes was prepared and evaluated for the ability to block dopamine, serotonin, and norepinephrine reuptake by their respective transporters. Compound 5b is a NET-selective inhibitor, 5c is a mixed NET- and SERT-selective inhibitor, while 11 is a SERT-selective inhibitor. PMID- 11909702 TI - Stereoselective synthesis of a C-glycoside analogue of N-Fmoc-serine beta-N acetylglucosaminide by Ramberg-Backlund rearrangement. AB - A C-glycoside analogue of N-Fmoc-serine beta-N-acetylglucosaminide 1 was synthesized stereoselectively from a sulfone derivative of serinol thio-N acetylglucosamide 8 using a Ramberg-Backlund rearrangement as a key step. PMID- 11909703 TI - Liquid-phase combinatorial synthesis of aminobenzimidazoles. AB - Liquid-phase synthesis of 2-arylaminobenzimidazoles using soluble polymer support strategy is first described. The key step to benzimidazole skeleton is achieved in the presence of diisopropylcarbodimide (DICDI) and isothiocyanates. A mechanism study for one-pot cyclodesulfurization is also investigated on the support. A wide range of benzimidazole derivatives is synthesized in excellent yield and purity just by simple wash and precipitation. PMID- 11909704 TI - Azapeptides as inhibitors of the hepatitis C virus NS3 serine protease. AB - Truncation and substitution SAR studies of azapeptide-based inhibitors of the Hepatitis C virus (HCV) NS3 serine protease have been performed. These azapeptides were designed from the HCV polyprotein's NS5A-NS5B trans cleavage junction and contained an azaamino acid residue at the P1 position. These azapeptides exhibited predominantly non-acylating, competitive inhibition, contrary to classical azapeptides. PMID- 11909705 TI - Novel potent antagonists of human neuropeptide Y-Y5 receptor. Part 4: tetrahydrodiazabenzazulene derivatives. AB - Novel tetrahydrodiazabenzazulene derivatives, designed from the lead compound 1 discovered by screening of our in-house chemical library, were prepared and found to be potent neuropeptide Y-Y5 (NPY-Y5) receptor antagonists. The structure activity relationships are described. Compounds 7 (FR240662) and 16 (FR252384) were especially attractive owing to their high affinities for the NPY-Y5 receptors, oral absorption and permeability to brain. PMID- 11909706 TI - Antitumor agents. Part 3: synthesis and cytotoxicity of new trans-stilbene benzenesulfonamide derivatives. AB - A new series of trans-stilbene benzenesulfonamide derivatives were designed and synthesized as potential antitumor agents. These new compounds were evaluated in the National Cancer Institute's 60 human tumor cell line in vitro screen. Compounds 9-13 were cytotoxic against several cell lines. Notably, two compounds, 9 and 12, demonstrated selective cytotoxic activity against BT-549 breast cancer (GI(50)=0.205 microM) and HT-29 colon cancer (GI(50)=0.554 microM), respectively. PMID- 11909707 TI - Noncovalent tripeptidic thrombin inhibitors incorporating amidrazone, amine and amidine functions at P1. AB - A series of noncovalent tripeptidic thrombin inhibitors incorporating amidrazone, amine and amidine functions at P1 was investigated. While the amidrazone and the amine series displayed limited oral absorption, the amidine series demonstrated generally good oral absorption and strong antithrombotic activity; the single digit picomolar K(i) achieved from this series is among the best yet reported. The present work highlights the benzamidine compound 11f (LB30812) that exhibits excellent overall profiles of potency, oral absorption and antithrombotic efficacy. PMID- 11909708 TI - The synthesis and characterization of BMS-204352 (MaxiPost) and related 3 fluorooxindoles as openers of maxi-K potassium channels. AB - 3-Aryl-3-fluorooxindoles can be efficiently synthesized in two steps by the addition of an aryl Grignard to an isatin, followed by treatment with DAST. Oxindole 1 (BMS-204352; MaxiPost) can be isolated using chiral HPLC or prepared by employing chiral resolution. Cloned maxi-K channels are opened by 1, which demonstrates a brain/plasma ratio >9 in rats. PMID- 11909710 TI - Optimization of the enzymatic synthesis of O-glycan core 2 structure by use of a genetic algorithm. AB - The enzymatic synthesis of Gal-beta 1,3[GlcNAc-beta 1,6]-GalNAc-alpha 1-OBn (core 2-Bn) using a multi-enzyme system consisting of a beta-galactosidase (EC 3.2.1.23) from bovine testes and a recombinant core 2 beta 1,6-GlcNAc transferase (C2GnT, EC 2.4.1.102) was empirically optimized by the use of a genetic algorithm. After variation of seven relevant parameters and performance of 56 experiments, two local maxima regarding the selection criteria could be found after four generations of optimization. The selectivity of core 2-Bn formation showed values up to 90%. PMID- 11909709 TI - A novel dicyanotriterpenoid, 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-onitrile, active at picomolar concentrations for inhibition of nitric oxide production. AB - New oleanane triterpenoids with various substituents at the C-17 position of 2 cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO) and methyl 2-carboxy-3,12 dioxooleana-1,9(11)-dien-28-oate were synthesized. Among them, 2-cyano-3,12 dioxooleana-1,9(11)-dien-28-onitrile shows extremely high inhibitory activity (IC(50)=1 pM level) against production of nitric oxide induced by interferon gamma in mouse macrophages. This potency is about 100 times and 30 times more potent than CDDO and dexamethasone, respectively. PMID- 11909711 TI - Reductive amination products containing naphthalene and indole moieties bind to melanocortin receptors. AB - Presumed pharmacophoric groups of melanocortin peptides (naphthalene, amino or guanidine, and indole moieties) were combined in mimetics molecules looking for their favorable location for activity at melanocortin (MC) receptors. Twenty-two compounds were prepared and tested. The best of these displayed micromolar affinities for the MC receptors. PMID- 11909712 TI - N-alkylaminoacids and their derivatives interact with melanocortin receptors. AB - Thirty four N-alkylaminoacids (Arg, Trp, Nal, Pro, Hyp, L and D) and derivatives were prepared by a process that included reductive alkylation of the amino function. Both solid phase and solution synthesis was used. Title substances displayed binding activity on melanocortin receptors MC(1,3-5) reaching the low micromolar range. PMID- 11909713 TI - Thalidomide and its analogues as cyclooxygenase inhibitors. AB - Thalidomide showed cyclooxygenase (COX)-1/2 inhibitory activity with a potency comparable to that of aspirin. Structural development studies of thalidomide resulted in potent COX-1/2 inhibitors, and COX-1-selective and COX-2-selective inhibitors. PMID- 11909714 TI - New thiazane and thiazolidine PNA monomers: synthesis, incorporation into PNAs and hybridization studies. AB - New constrained PNA monomers containing a substituted thiazolidine or a thiazane ring were synthesized and incorporated in the center of a 9-mer homothymine PNA. The PNA/DNA hybrids stability was studied by UV-melting experiments which showed that the presence of the modified unit destabilizes the PNA/DNA triplexes. PMID- 11909715 TI - Squaric acid derivatives as VLA-4 integrin antagonists. AB - SAR studies aimed at improving the rate of clearance by the incorporation of a 3,4-diamino-3-cyclobutene-1,2-dione group as an amino acid isostere in a series of VLA-4 integrin antagonists are described. PMID- 11909716 TI - Covalent analogues of DNA base-pairs and triplets. Part 2: synthesis and cytostatic activity of bis(purin-6-yl)acetylenes,-diacetylenes and related compounds. AB - The title bis(purin-6-yl)acetylenes, -diacetylenes, -ethylenes and -ethanes were prepared as covalent base-pair analogues starting from 6-ethynylpurines and 6 iodopurines by cross-coupling and homo-coupling reactions and hydrogenations. The bis(purin-6-yl)acetylenes and -diacetylenes exhibited significant cytostatic activity in vitro (IC(50)=0.4-1.0 micromol/l). PMID- 11909717 TI - New aromatase inhibitors. Synthesis and inhibitory activity of pyridinyl substituted flavanone derivatives. AB - Two (E)-pyridinyl-substituted flavanone derivatives were synthesized and UV irradiation of these compounds afforded a Z-enriched mixture. These products were tested for their ability to inhibit the cytochrome P450 aromatase. It was observed that the introduction of a pyridinylmethylene group at carbon 3 on flavanone nucleus led to a significant increase of aromatase inhibitory effect. Moreover, configuration had a substantial influence on the aromatase inhibitory activity since (E)-isomers were found to be more active than (Z)-isomers. PMID- 11909718 TI - Design, synthesis and in vitro evaluation of potent, novel, small molecule inhibitors of plasminogen activator inhibitor-1. AB - We have synthesized and evaluated a series of tetramic acid-based and hydroxyquinolinone-based inhibitors of plasminogen activator inhibitor-1 (PAI-1). These studies resulted in the identification of several compounds which showed excellent potency against PAI-1. The design, synthesis and SAR of these compounds are described. PMID- 11909719 TI - Synthesis and preliminary biological assessments of RGD bearing biocompatible telomers. AB - Work reported herein deals with the synthesis and preliminary biological assessments of a new class of biocompatible telomeric carriers bearing peptidic RGDSK sequences as tumor cell targeting and tyrosine moieties labelled with 125I as in vivo probe. The radioactivity levels obtained in several tissues, after the intravenous injections of these telomers in mice bearing grafted B16 syngenic melanoma showed that the addition of a RGD residue to a telomeric structure confers it an increased affinity for the highly vascularized zone surrounding the tumor. PMID- 11909720 TI - Benzoindoloquinolines interact with DNA tetraplexes and inhibit telomerase. AB - Telomeric G-rich single-stranded DNA can adopt a G-tetraplex structure which has been shown to inhibit telomerase activity. We have examined benzoindoloquinolines derivatives for their ability to stabilize an intramolecular G-quadruplex. The increase in T(m) value of the G-quadruplex was associated with telomerase inhibition in vitro. PMID- 11909721 TI - A 3,4-epoxypiperidine structure as a novel and simple DNA-cleavage unit. AB - Based on the 4-hydroxy-1-azabicyclo[3.1.0]hexane structure of azinomycin, a 3,4 epoxypiperidine structure was designed as a novel and simple alkylating molecular unit, and some 3,4-epoxypiperidine derivatives were found to show DNA-cleavage activity, the structural requirements for which were revealed. PMID- 11909722 TI - Unprecedented overmetabolism of a porphyrinogen substrate by coproporphyrinogen oxidase. AB - Harderoporphyrinogen-I is metabolized by avian hemolysate preparations of coproporphyrinogen oxidase to give a trivinylic product; this unprecedented 'overmetabolism' of the porphyrinogen substrate provides strong support for a proposed model of the active site of this poorly understood enzyme. PMID- 11909723 TI - New highly active taxoids from 9 beta-dihydrobaccatin-9,10-acetals. AB - To synthesize new highly active taxoids, we designed and synthesized 9 beta dihydro-9,10-acetal taxoids. In vitro study of these analogues clearly showed them to be more potent than docetaxel. PMID- 11909724 TI - Sulfide analogues as potent and selective M(2) muscarinic receptor antagonists. AB - We have discovered highly potent, selective sulfide M(2) receptor antagonists with low molecular weight and different structural features compared with our phase I clinical candidate Sch 211803. Analogue 30 showed superior M(2) receptor selectivity profile over Sch 211803. More importantly, this study provided new leads for the discovery of M(2) receptor antagonists as potential drug candidates. PMID- 11909725 TI - Benzobicyclooctanes as novel inhibitors of TNF-alpha signaling. AB - A novel series of TNF-alpha inhibitors based on a benzobicyclooctane scaffold is reported. The compounds display good potency in inhibiting TNF-alpha induced apoptosis and NF kappa B activation. Additionally, they are selective for TNF alpha as they do not inhibit apoptosis induced by soluble Fas ligand. The compounds described here can act as leads for future medicinal chemistry efforts and may also be useful tools for elucidating the TNF-alpha signaling pathway. PMID- 11909726 TI - 5-Phosphonomethylquinoxalinediones as competitive NMDA receptor antagonists with a preference for the human 1A/2A, rather than 1A/2B receptor composition. AB - NMDA antagonists derived from 5-phosphonomethyl-1,4-dihydroquinoxaline-2,3-dione (3a) are potent anticonvulsant agents, and display strong protective effects in the electroshock-induced convulsion assay in mice. Their preference for the human NMDAR 1A/2A over 1A/2B subunit composition was optimized, leading to (1RS,1'S) PEAQX (9r), which shows a >100-fold selectivity. PMID- 11909727 TI - Spirocyclic nonpeptide glycoprotein IIb-IIIa antagonists. Part 3: synthesis and SAR of potent and specific 2,8-diazaspiro[4.5]decanes. AB - The synthesis and biological activity of analogues containing spiro piperidinylpyridine and pyrrolidinylpyridine templates are described. The potent activity of these compounds as platelet aggregation inhibitors demonstrates the utility of the spiro structures as central template for nonpeptide RGD mimics. PMID- 11909728 TI - Structural approach of the mechanism of inhibition of alpha-chymotrypsin by coumarins. AB - A pharmacophore associated to the inhibiton of alpha-chymotrypsin has been built based on the structural and electronic characterization of a series of coumarin derivatives. PMID- 11909729 TI - First and unexpected synthesis of macrocyclic cyclophane-based unusual alpha amino acid derivatives by phosphazene base without high dilution conditions. AB - First synthesis of a macrocylic cyclophane-based unusual alpha-amino acid derivative 11 by coupling of ethyl isocyanoacetate with 1,2-bis(4 bromomethylphenyl)ethane under phase-transfer catalysis (PTC) conditions. Phosphazene base such as 2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro 1,3,2-diazaphosphorine (BEMP) is useful to improve the yield of cyclophane derivative without high dilution conditions. PMID- 11909730 TI - The synthesis and structure-activity relationships of 1,3-diaryl 1,2,4-(4H) triazol-5-ones: a new class of calcium-dependent, large conductance, potassium (maxi-K) channel opener targeted for urge urinary incontinence. AB - A series of 1,3-diaryl 1,2,4-(4H)-triazol-5-ones was prepared and shown by electrophysiological analysis to activate a cloned maxi-K channel mSlo (or hSlo) expressed in Xenopus laevis oocytes. The effects of these structurally novel maxi K channel openers on bladder contractile function were studied in vitro using isolated rat bladder strips pre-contracted with carbachol. Several 1,3-diaryl 1,2,4-(4H)-triazol-5-one derivatives were found to be potent smooth muscle relaxants but this activity did not completely correlate with maxi-K channel opening. PMID- 11909731 TI - Synthesis of base-modified dihydropacidamycins. AB - We describe in this paper the synthesis of 1,2-di-O-acetyl-5-azido-3,5-dideoxy alpha,beta-L-arabinofuranose, a carbohydrate donor that was used for the synthesis of 1-(5'-amino-3',5'-dideoxy-alpha-L-arabinofuranosyl)uracil, the nucleoside found in dihydropacidamycin D. The carbohydrate donor was also used for the synthesis of a set of new nucleosides that were introduced in new dihydropacidamycins. These compounds were tested for biological activity, and the results showed that uracil is the only base recognized by MraY. PMID- 11909732 TI - Novel estimation of lipophilic behaviour of polychlorinated biphenyls. AB - A novel method has been developed for the estimation of lipophilic behaviour of polychlorinated biphenyls (PCBs) using the recently introduced Padmakar-Ivan (PI) index. The results obtained are compared with the earlier reported Abraham method. The statistical analyses showed that the proposed method based on the PI index is quite useful. PMID- 11909733 TI - Beta-carbolines as specific inhibitors of cyclin-dependent kinases. AB - Harmine (3), 7-fluoro-1-methyl beta-carboline (35) and 1-(5-methyl-imidazol-4-yl) beta-carboline (41) were potent and specific inhibitors of cyclin-dependent kinases. The degree of aromaticity of the tricyclic ring and the positioning of substituents are important for inhibitory activity. While most beta-carbolines inhibited CDK2 and CDK5 to the same extent, selective inhibition against CDK2 was observed in 1-(2-chlorophenyl)- (12), 1-(2-fluorophenyl)- (15), and 1-(2-chloro-5 nitrophenyl)- (28) beta-carbolines. PMID- 11909735 TI - Toxoplasma gondii: in vivo expression of BAG-5 and cyst formation is independent of TNF p55 receptor and inducible nitric oxide synthase functions. AB - Wild type, TNFRp55(-/-), iNOS(-/-) and IFN-gamma(-/-) mice were infected with Toxoplasma gondii strain ME-49, and the central nervous system (CNS), lungs, liver, spleen, heart and kidneys were examined for the presence of parasites expressing tachyzoite-specific (SAG-1) and bradyzoite-specific (BAG-5) antigens. During the acute phase of infection, the peripheral organs, but not the CNS, of the IFN-gamma(-/-) mice are heavily parasitized by tachyzoites and there are no signs of parasites expressing BAG-5. In contrast, the tissues from TNFRp55(-/-) and inducible nitric oxide synthase (iNOS)(-/-) mice, mainly the CNS, presented high numbers of parasites expressing SAG-1 and/or BAG-5. Tachyzoite transformation into bradyzoite and cyst development was shown to be normal in the tissues from TNFRp55(-/-) and iNOS(-/-) mice, as indicated by the high numbers of BAG-5/PAS positive cysts. Consistently, reactivation of infection in IFN-gamma(-/ ) mice was rapid and characterized by a dramatic increase in SAG-1, contrasting with slow course in the TNFRp55(-/-) or iNOS(-/-) mice associated with a relatively small increase in SAG-1- and/or BAG-5-positive parasites. In conclusion, our results suggest that the control of multiplication of tachyzoites is largely dependent on endogenous IFN-gamma with partial involvement of TNFRp55 and iNOS. In contrast, induction of BAG-5 expression and cyst formation during toxoplasmosis seems to be dependent on IFN-gamma, but independent of TNFRp55 and iNOS functions. PMID- 11909736 TI - A role for Toxoplasma gondii type 1 ser/thr protein phosphatase in host cell invasion. AB - Host cell invasion by Toxoplasma gondii tachyzoites relies on many coordinated processes. The tachyzoite participates in invasion by providing an actomyosin dependent force driving it into the nascent parasitophorous vacuole as well as by releasing molecules which contribute to the vacuole membrane. Exposure to type 1/2A protein phosphatase inhibitors, okadaic acid (OA) or tautomycin significantly impairs tachyzoite invasiveness. Furthermore, the tachyzoite extract contains a biochemically active type 1, but not a type 2A, serine threonine protein phosphatase, which is immunologically related to eukaryotic phosphatase type 1 catalytic subunit. When tachyzoite extracts are incubated with a monoclonal antibody reactive to human type 1 catalytic subunit, other T. gondii molecules are coprecipitated among which one competes with the inhibitory toxin OA. Finally, in vitro phosphate labelling assays indicate that the biochemically characterized PP1 activity controls the phosphorylation of several proteins. Taken together, these data strongly suggest that the type 1 phosphatase activity detected in invasive tachyzoites is implicated in the control of the host cell invasion process. PMID- 11909737 TI - Adherence of Flavobacterium psychrophilum on the body surface of the ayu Plecoglossus altivelis. AB - Bacterial cold water disease in the ayu Plecoglossus altivelis caused by Flavobacterium psychrophilum is a serious problem in the Japanese freshwater culture industry. The distribution and activity of this bacterium on the body surface of the ayu in the infection process was investigated. The survival of F. psychrophilum in tap water showed that this bacterium might sustain its infectivity for 24 h. In an experimental infection, juvenile ayu were immersed in water containing 10(8.9) CFU/ml F. psychrophilum, and the progressing infection was followed by scanning electron microscopy during a 24-h period. This bacterium was observed in the ayu for 24 h adhering to the lower jaw and caudal peduncle, where the epidermis tissue was collapsed. This study showed that bacterial suspension in water sustains the activity of this bacterium. F. psychrophilum attaches especially to the jaw and caudal peduncle, growing at these sites, collapsing the dermal structure and invading the tissues. PMID- 11909738 TI - A PCR-ELISA for detecting Shiga toxin-producing Escherichia coli. AB - A sensitive and specific PCR-ELISA was developed to detect Escherichia coli O157:H7 and other Shiga toxin-producing E. coli (STEC) in food. The assay was based on the incorporation of digoxigenin-labeled dUTP and a biotin-labeled primer specific for Shiga toxin genes during PCR amplification. The labeled PCR products were bound to streptavidin-coated wells of a microtiter plate and detected by an ELISA. The specificity of the PCR was determined using 39 bacterial strains, including STEC, enteropathogenic E. coli, E. coli K12, and Salmonella. All of the STEC strains were positive, and non-STEC organisms were negative. The ELISA detecting system was able to increase the sensitivity of the PCR assay by up to 100-fold, compared with a conventional gel electrophoresis. The detection limit of the PCR-ELISA was 0.1-10 CFU dependent upon STEC serotypes, and genotypes of Shiga toxins. With the aid of a simple DNA extraction system, PrepMan, the PCR-ELISA was able to detect ca. 10(5) CFU of STEC per gram of ground beef without any culture enrichment. The entire procedure took about 6 h. Because of its microtiter plate format, PCR-ELISA is particularly suitable for large-scale screening and compatible with future automation. PMID- 11909739 TI - Cerebral malaria: the contribution of studies in animal models to our understanding of immunopathogenesis. AB - Cerebral malaria is a serious and often fatal complication of Plasmodium falciparum infections. The precise mechanisms involved in the onset of neuropathology remain unknown, but parasite sequestration in the brain, metabolic disturbances and host immune responses are all thought to be involved. This review outlines the current state of knowledge of cerebral disease in humans, and discusses the contribution of studies of animal models to elucidation of the underlying mechanisms. PMID- 11909740 TI - HIV-1 and the brain: connections between HIV-1-associated dementia, neuropathology and neuroimmunology. AB - AIDS patients frequently exhibit neurological disorders due to the neurotoxic events that result from HIV-1 and/or opportunistic infections in the brain. This review examines recent clinical findings related to HIV-1-associated dementia, and outlines current areas of basic research that may clarify how HIV-1 associated encephalopathy produces clinical symptoms of brain dysfunction. PMID- 11909741 TI - The gut microflora and intestinal epithelial cells: a continuing dialogue. AB - The mammalian intestinal epithelium effectively performs its physiological functions in a microbe-rich environment, while the prokaryotic population thrives amidst efficient cellular defenses. Recent delineation of the mechanisms by which bacteria communicate with their eukaryotic hosts and the cellular sites that microbial signals act in may shed light on these complex interactions. PMID- 11909742 TI - Use of Lactobacillus to prevent infection by pathogenic bacteria. AB - This review focuses on the use and potential of Lactobacillus to prevent infections of the urogenital and intestinal tracts. The presence and dominance of Lactobacillus in the vagina is associated with a reduced risk of bacterial vaginosis and urinary tract infections. The mechanisms appear to involve anti adhesion factors, by-products such as hydrogen peroxide and bacteriocins lethal to pathogens, and perhaps immune modulation or signaling effects. The instillation of Lactobacillus GR-1 and B-54 or RC-14 strains into the vagina has been shown to reduce the risk of urinary tract infections, and improve the maintenance of a normal flora. Ingestion of these strains into the gut has also been shown to modify the vaginal flora to a more healthy state. In addition, these strains inhibit the growth of intestinal, as well as urogenital pathogens, colonize the gut and protect against infections as shown in mice. Other probiotic strains, such as Lactobacillus GG, have been shown to prevent and treat gastroenteritis caused by rotavirus and bacteria. Given that lactobacilli are not the dominant commensals in a gut which comprises around 10(10) organisms, much work is still needed to define the mechanisms whereby GR-1, RC-14, GG and other strains contribute to health restoration and maintenance. Such critically important studies will require the medical science community to show a willingness to turn away from pharmaceutical remedies as the only solution to health and disease. PMID- 11909743 TI - Outer membrane proteins: key players for bacterial adaptation in host niches. AB - Outer membrane proteins (OMPs) of Gram-negative bacteria have diverse functions and are directly involved in the interaction with various environments encountered by pathogenic organisms. Thus, OMPs represent important virulence factors and play essential roles in bacterial adaptation to host niches, which are usually hostile to invading pathogens. Understanding the structure and functions of bacterial OMPs will facilitate the design of antimicrobial drugs and vaccines. In this paper, we will present a brief review on OMPs that contribute to bacterial adaptive responses including iron uptake, antimicrobial peptide resistance, serum resistance, and drug/bile resistance. PMID- 11909744 TI - Proteins in the chlamydial inclusion membrane. AB - The chlamydiae are obligate intracellular pathogens that occupy a nonacidified vacuole (the inclusion) during their entire developmental cycle. Several proteins have recently been identified that are localized to the inclusion membrane. The following is a discussion of how inclusion membrane proteins might participate in the chlamydial developmental process. PMID- 11909745 TI - La Crosse virus: replication in vertebrate and invertebrate hosts. AB - La Crosse virus is maintained in a cycle involving mosquitoes and small mammals. Vertebrate cell infection is generally cytolytic; vector cell infection results in persistent infection. Features of La Crosse virus replication that may permit the virus to traffic between vector and vertebrate hosts and condition different infection outcomes are described. PMID- 11909746 TI - The natural resistance-associated macrophage protein 1 Slc11a1 (formerly Nramp1) and iron metabolism in macrophages. AB - Slc11a1 (solute carrier family 11 member 1) (formerly Nramp1) modulation of iron metabolism in macrophages plays an important role in early phase macrophage activation, and therefore host innate immunity. This review focuses on the role of Nramp1 in intramacrophage iron metabolism, with emphasis on the two prevailing mechanisms of Nramp1 modulation of iron metabolism in macrophages. PMID- 11909747 TI - Cathelicidins: microbicidal activity, mechanisms of action, and roles in innate immunity. AB - Antimicrobial peptides are important host-defense molecules of innate immunity. Cathelicidins are a diverse family of potent, rapidly acting and broadly effective antimicrobial peptides, which are produced by a variety of cells. This review examines the classification, antimicrobial spectrum, mechanism of action, and regulation of cathelicidins. PMID- 11909748 TI - Antibodies against gangliosides: a link between preceding infection and immunopathogenesis of Guillain-Barre syndrome. AB - Autoantibodies against gangliosides GM1 and GQ1b, characteristic cell surface glycolipids of the nervous system, are present in specific clinical types of GuillainBarre syndrome (GBS). Close associations of anti-GM1 with acute motor axonal neuropathy, and of anti-GQ1b with Miller Fisher syndrome, strongly suggest that these antibodies contribute to neuropathy pathogenesis. Immune responses against gangliosides are suspected to originate as a result of molecular mimicry between gangliosides and lipopolysaccharides of Campylobacter jejuni, the most frequent infectious trigger of GBS. Thus, antibodies against gangliosides may link C. jejuni infection with the precipitation of neurological disease. PMID- 11909749 TI - The epidemiology of variant Creutzfeldt-Jakob disease in Europe. AB - Variant Creutzfeldt-Jakob disease is one of a family of neurodegenerative diseases, first diagnosed in 1996. Scientific evidence strongly supports the hypothesis that it is acquired through consumption of bovine spongiform encephalopathy-infected meat. The majority of cases have been diagnosed in the UK in young individuals, with an excess of cases in the north and a significant cluster of cases in Leicestershire. Many uncertainties in its biology and epidemiology, in particular the length of the incubation period, make predictions of any future epidemic difficult. Studies are currently under way to obtain more precise estimates of the prevalence of asymptomatic infection through testing tonsil and appendix tissues for the abnormal prion protein. PMID- 11909750 TI - A simplified and rapid method for scoring micronucleated erythrocytes in human blood. PMID- 11909751 TI - Enumeration of micronucleated CD71-positive human reticulocytes with a single laser flow cytometer. AB - The extreme rarity of micronucleated reticulocytes (RETs) in the peripheral blood of non-splenectomized humans has precluded facile enumeration of these cells, as well as evaluation of this endpoint as an index of cytogenetic damage. In this report, we describe a high-throughput, single-laser flow cytometric system for scoring the incidence of micronuclei (MN) in newly formed human RETs. The procedure is based on an immunochemical reagent that differentially labels the most immature fraction of RETs from mature erythrocytes based on the expression level of the transferrin receptor (also known as CD71). The resolution of four erythrocyte populations (young RETs and mature erythrocytes, with and without MN) was achieved for human blood cells treated with phycoerythrin-conjugated anti CD71, RNase, and either SYTOX Green or SYBR Green I nucleic acid dyes. Anti glycophorin A labeling of erythroid cells (CyChrome conjugate) was also incorporated into the staining procedure to ensure that debris or other potential artifacts did not adversely impact the analyses. Instrument calibration procedures utilizing malaria-infected rodent erythrocytes were also developed, and are described. Using this analytical system, blood samples from 10 healthy non-splenectomized human volunteers were analyzed for micronucleus frequencies with a single-laser flow cytometer. Average micronucleus frequencies in the mature and most immature fraction of RETs were 0.016 and 0.19%, respectively. Blood samples from three healthy splenectomized volunteers were also evaluated. As expected, these samples exhibited higher micronucleus frequencies in the mature subset of erythrocytes (range 0.03-0.18%). The resulting data suggest that MN can be quantified in human erythrocyte populations with a single-laser flow cytometer, and that the frequency of MN cells in the youngest reticulocyte population approaches values expected in the absence of splenic selection against MN-erythrocytes. This high throughput system is potentially important for evaluating the value of the micronucleated reticulocyte endpoint as an index of chromosome breakage and/or chromosome segregational abnormalities in human populations. PMID- 11909752 TI - Transformation of mutagenic aromatic amines into non-mutagenic species by alkyl substituents. Part II: alkylation far away from the amino function. AB - Alkyl and trifluoromethyl derivatives of 4-aminobiphenyl (1) (4ABP) and 2 aminofluorene (7) (2AF) were synthesised and assayed for mutagenicity using Salmonella typhimurium tester strains TA98 and TA100 with and without the addition of S9 mix. Modification of 1 was achieved by attachment of alkyl groups (methyl, ethyl, iso-propyl, n-butyl, tert-butyl) and a trifluoromethyl group (CF(3)) in the 4'-position, the 3'-position (Me, CF(3)) and the 3'-, 5'-positions (DiMe, DiCF(3)). Compound 7 was modified by introduction of alkyl groups (methyl, tert-butyl, adamantyl) and a trifluoromethyl group (CF(3)) in the 7-position. The derivatives of 1 and 7 show for groups with growing steric demand decreased mutagenic activity. The bulkiest groups (CF(3), tert-butyl and adamantyl) induce the strongest effects on the mutagenicity. It was even possible to eliminate the mutagenicity of 1 and 7 by introduction of such substituents. In the last part of the work, we compared the experimental mutagenicities with calculated values derived from QSAR correlations. Our findings show that the predictions for aromatic amines with bulky substituents were generally too high. The strongest deviations were observed in the case of the CF(3)-, tert-butyl- and the adamantyl group. Only the parent compounds and derivatives with small alkyl groups were predicted well. These investigations show that "large" substituents have an influence on the mutagenicity caused by their steric demand. To predict the correct mutagenicities of such compounds, it is necessary to introduce steric parameters in the respective QSAR equations which will be done in a forthcoming paper. PMID- 11909753 TI - Evaluation of the antigenotoxic potential of monomeric and dimeric flavanols, and black tea polyphenols against heterocyclic amine-induced DNA damage in human lymphocytes using the Comet assay. AB - The polyphenolic dimers, epicatechin-4beta-8-catechin (B1), epicatechin-4beta-8 epicatechin (B2), catechin-4beta-8-catechin (B3), catechin-4beta-8-epicatechin (B4), and the gallate ester epicatechin-4beta-8-epicatechin gallate (B'2G) were isolated from grape seeds, and theaflavins and theafulvins from black tea brews. The ability of these naturally-occurring polyphenols to afford protection against the genotoxicity of the heterocyclic amine 3-amino-1-methyl-5H-pyrido[4,3 b]indole (Trp-P-2) was compared with that of the monomeric tea flavanols, (+) catechin (C), (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-) epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG). Genotoxic activity was evaluated in human peripheral lymphocytes using the Comet assay. At the concentration range of 1-100 microM, neither the monomeric nor the dimeric flavanols prevented the lymphocyte DNA damage induced by Trp-P-2. In contrast, both of the black tea polyphenols, theafulvins and theaflavins, at a dose range of 0.1-0.5 mg/ml, prevented, in a concentration-dependent manner, the DNA damage elicited by Trp-P-2. Finally, neither the monomeric and dimeric polyphenols (100 microM) nor the theafulvins and theaflavins (0.5mg/ml) caused any DNA damage in the human lymphocytes. These studies illustrate that black tea theafulvins and theaflavins, if absorbed intact, may contribute to the anticarcinogenic potential associated with black tea intake. PMID- 11909754 TI - Micronuclei in cervical smears and peripheral blood lymphocytes from women with and without cervical uterine cancer. AB - Cervical cancer represents the second most common malignant neoplasia in women world-wide. In Mexico, cervical cancer is the most common female malignancy. It has been recently seen an increased frequencies of micronuclei (MN) lymphocytes and cervical epithelial cells of cervical cancer patients. The aim of this hospital-based unmatched case-control study was to investigate the association between progressive stages in development of cervical cancer and frequency of micronucleated cells in the cervical epithelium and peripheral lymphocytes of 40 women, grouped by disease stage. Women at the Obstetrics and Gynecology Hospital of the Instituto Mexicano del Seguro Social (IMSS) in Monterrey, Mexico were diagnosed and classified on the bases of the Papanicolaou (PAP) smear and colposcopy/biopsy into control, low-grade squamous intraepithelial lesions (LGSIL), high-grade squamous intraepithelial lesions (HGSIL), and invasive groups. Analysis of the MN data in both cell types revealed (a) homogeneity among women within each of the four groups with regard to MN frequency, (b) in general, a correlation between MN frequency and grade of cervical lesion, and (c) a positive linear trend between the MN frequency and increased cervical cancer risk. In conclusion, we suggest that MN are a useful biomarker of cancer risk. Nonetheless, these results should be validated by other researchers. PMID- 11909755 TI - Mutagenicity of aristolochic acid in the lambda/lacZ transgenic mouse (MutaMouse). AB - Aristolochic acid (AA) is found in a plant that causes urothelial carcinomas in patients with Chinese herb nephropathy (CHN). To evaluate the in vivo mutagenicity of AA, we analysed the mutant frequency (MF) in the lacZ and cII gene of 10 organs of the lambda/lacZ transgenic mouse (MutaMouse) after intragastric treatment with AA (15mg/kg per week x 4). Simultaneously, the clastogenicity of AA was evaluated by the peripheral blood micronucleus assay. The nature of the mutations induced by AA was revealed by the sequence analysis of the cII gene, which is also a phenotypically selectable marker in the lambda transgene. MFs in the target organs-forestomach, kidney, and bladder of AA treated mice were significantly higher than those of control mice (forestomach 33 and 15-fold; kidney 10- and 9-fold; bladder 16- and 31-fold, for the lacZ and cII, respectively). The MFs in non-target organs, except the colon, showed only slight increases. Sequence analysis of cII mutants in target organs revealed that AA induced mainly A:T to T:A transversions whereas G:C to A:T transitions at CpG sites predominated among spontaneous mutations. These results suggested that AA, which is activated by cytochrome P450 and peroxidase to form cyclic nitrenium ions that bind to deoxyadenine, caused the A to T transversions in the target organs of mice. PMID- 11909757 TI - The genotoxicity of priority polycyclic aromatic hydrocarbons in complex mixtures. AB - Risk assessment of complex environmental samples suffers from difficulty in identifying toxic components, inadequacy of available toxicity data, and a paucity of knowledge about the behavior of geno(toxic) substances in complex mixtures. Lack of information about the behavior of toxic substances in complex mixtures is often avoided by assuming that the toxicity of a mixture is simply the sum of the expected effects from each mixture component, i.e. no synergistic or antagonistic interactions. Although this assumption is supported by research investigating non-genotoxic end-points, the literature describing the behavior of genotoxic substances in complex mixtures is sparse and, occasionally, contradictory. In this study, the results of polycyclic aromatic hydrocarbon (PAH) analyses on freshwater bivalves were used to prepare realistic mixtures containing up to 16 PAHs. The SOS genotoxicity of the mixtures and each component were then assessed in an effort to evaluate the additivity of PAH genotoxicity. At nominal PAH concentrations above 1 microg/ml, observed genotoxic responses were far lower than those predicted under the assumption of additivity. At nominal concentrations below 0.75 microg/ml, differences are smaller and occasionally negligible, indicating that the genotoxicity of unsubstituted homocyclic PAHs is additive or slightly less than additive. Other researchers who have investigated the mutagenicity, carcinogenicity, and DNA binding activity of mixtures containing unsubstituted homocyclic PAHs have also reported additive effects. Therefore, the mutagenic risk posed by simple, well-characterized mixtures of priority PAHs can reasonably be estimated as the sum of the risks posed by the mixture components. Current data indicate that less-than-additive effects likely result from saturation of metabolic pathways needed to activate mutagenic PAHs. PMID- 11909756 TI - Dinitropyrenes induce gene mutations in multiple organs of the lambda/lacZ transgenic mouse (Muta Mouse). AB - Dinitropyrenes (DNPs), 1,3-, 1,6- and 1,8-dinitropyrene, are carcinogenic compounds found in diesel engine exhaust. DNPs are strongly mutagenic in the bacterial mutation assay (Ames test), mainly inducing frameshift type mutations. To assess mutagenicity of DNPs in vivo is important in evaluating their possible involvement in diesel exhaust-induced carcinogenesis in human. For this purpose, we used the lambda/lacZ transgenic mouse (Muta Mouse) to examine induction of mutations in multiple organs. A commercially available mixture of DNPs (1,3-, 1,6 , 1,8-, and unidentified isomer (s) with a content of 20.2, 30.4, 35.2, and 14.2%, respectively) was injected intragastrically at 200 and 400mg/kg once each week for 4 weeks. Seven days after the final treatment, liver, lung, colon, stomach, and bone marrow were collected for mutation analysis. The target transgene was recovered by the lambda packaging method and mutation of lacZ gene was analyzed by a positive selection with galE(-) E. coli. In order to determine the sequence alterations by DNPs, the mutagenicity of the lambda cII gene was also examined by the positive selection with hfl(-) E. coli. Since cII gene (294bp) is much smaller than the lacZ (3024bp), it facilitated the sequence analysis. Strongest increases in mutant frequencies (MFs) were observed in colon for both lacZ (7.5x10(-5) to 43.3x10(-5)) and cII (2.7x10(-5) to 22.5x10(-5)) gene. Three-four-fold increases were observed in stomach for both genes. A statistically significant increase in MFs was also evident in liver and lung for the lacZ gene, and in lung and bone marrow for the cII gene. The sequence alterations of the cII gene recovered from 37 mutants in the colon were compared with 50 mutants from untreated mice. Base substitution mutations predominated for both untreated (91%) and DNP-treated (84%) groups. The DNPs treatment increased the incidence of G:C to T:A transversion (2-43%) and decreased G:C to A:T transitions (70-22%). The G:C to T:A transversions, characteristic to DNPs treatment, is probably caused by the guanine-C8 adduct, which is known as a major DNA-adduct induced by DNPs, through an incorporation of adenine opposite the adduct ("A"-rule). The present study showed a relevant use of the cII gene as an additional target for mutagenesis in the Muta Mouse and revealed a mutagenic specificity of DNPs in vivo. PMID- 11909758 TI - Genotoxicological characterisation of complex mixtures. Genotoxic effects of a complex mixture of perhalogenated hydrocarbons. AB - Toxicological and genotoxicological investigation of complex mixtures is one of the main focus of the recent research in toxicology. Testing complex mixtures present a formidable scientific problem since most recently available toxicological data has been obtained from single substance studies and is not simply transferable to mixtures of chemicals. Although there are no special strategies and standard protocols available for determining toxic and genotoxic effects of complex mixtures, the fundamental concepts of evaluation are the same as those for single substances. The focus of interest of the submitted paper is the genotoxicological characterisation of a complex mixture of mostly perhalogenated hydrocarbons which is generated as a waste product from the plasma etching process in the semiconductor industry. By use of several in vitro test systems (comet assay and micronucleus test), the clastogenic potency of the mixture was tested in various human cell types (lymphocytes and normal bronchial epithelial cells) and in rat hepatocytes. Results demonstrated that the complex perhalogenated hydrocarbons mixture causes DNA single-strand breaks and micronuclei formation, and direct concentration-to-effect correlations were proved in all experiments. The presence of an external metabolising system (S9 mix from rat hepatocytes) in human cell culture systems did not cause any change of the observed effects when compared to experiments performed in the absence of the S9 mix. Therefore, we conclude that the mixture acts as direct genotoxicant and that there is no detoxification by the external enzyme system.Further, convincing and reproducible results of the in vitro comet assay and the micronucleus assay in primary human cell cultures indicated these tests may be utilized for the genotoxicological analyses of complex mixtures with concern to human health hazard. PMID- 11909759 TI - Effects of heavy metal contamination of soils on micronucleus induction in Tradescantia and on microbial enzyme activities: a comparative investigation. AB - The aim of this study was to investigate correlation between genotoxic effects and changes of microbial parameters caused by metal contamination in soils. In total, 20 soils from nine locations were examined; metal contents and physicochemical soil parameters were measured with standard methods. In general, a pronounced induction of the frequency of micronuclei (MN) in the Tradescantia micronucleus (Trad-MN) assay was seen with increasing metal concentration in soils from identical locations. However, no correlations were found between metal contents and genotoxicity of soils from different locations. These discrepancies are probably due to differences of the physicochemical characteristics of the samples. Also, the microbial parameters depended on the metal content in soils from identical sampling locations. Inconsistent responses of the individual enzymes were seen in soils from different locations, indicating that it is not possible to define a specific marker enzyme for metal contamination. The most sensitive microbial parameters were dehydrogenase and arylsulfatase activity, biomass C, and biomass N. Statistical analyses showed an overall correlation between genotoxicity in Tradescantia on the one hand and dehydrogenase activity, biomass C, and the metabolic quotient on the other hand. In conclusion, the results of the present study show that the Trad-MN assay is suitable for the detection of genotoxic effects of metal contamination in soils and furthermore, that the DNA-damaging potential of soils from different origin cannot be predicted on the basis of chemical analyses of their metal concentrations. PMID- 11909760 TI - Mutagenicity of sediments along the Po River and genotoxicity biomarkers in fish from polluted areas. AB - We monitored the mutagenicity of extracts of sediment fine particles collected, both in the cold season and in the hot season, from 10 reaches along the Po River, the main Italian watercourse. Each sample was representative of several kilometers of river stretch. At sub-toxic doses, the samples were not mutagenic to the Salmonella typhimurium his(-) strains TA98, TA100 and TA102, irrespective of the presence of S9 mix. However, they induced a mutagenic response in YG1024, which is typically reverted by frameshift mutagens that are metabolized in bacteria via acetyl-CoA:N-hydroxylamine O-acetyltransferase. Mutagenicity of sediments was higher during the cold season and had a spatial distribution consistent with the occurrence of pollution sources and confluence with polluted tributaries. Nevertheless, in the final stretch, near the Po delta into the Adriatic Sea, mutagenicity of sediments was low, comparable to that detected in the Po proximal reach, not far away from its springs. Genotoxicity biomarkers were evaluated in three cyprinid species, the "Italian nase" (Chondrostoma soetta), chub (Leuciscus cephalus), and barbel (Barbus plebejus), captured upstream and downstream of the confluence of a polluted tributary (Lambro River) with the Po River. There was no difference between the two areas concerning concentrations of fluorescent aromatic compounds in fish bile while, after metabolic activation, the bile of fish caught from the more polluted area became mutagenic to YG1024. Moreover, the levels of adducts to liver DNA were significantly higher in L. cephalus caught from the more polluted area, and the increase of micronucleated erythrocyte frequency was borderline to statistical significance, but only in C. soetta. Thus, certain biomarkers of exposure and effect in fish, as assessed under field conditions, correlate with the pollution of river sediments by mutagenic compounds. PMID- 11909761 TI - Chromosomal aberrations under basal conditions and after treatment with X-ray in human lymphocytes as related to the GSTM1 genotype. AB - The frequency of chromosomal aberrations (CAs) was evaluated in blood lymphocytes from 18 healthy subjects. Basal CA frequencies were not significantly different in GSTM1 positive and GSTM1 null subjects (P>0.05), whereas they were considerably higher in smokers than in non-smokers. After 1 Gy dose of X-ray challenge of blood samples, CA frequencies were significantly higher in GSTM1 null subjects, compared to GSTM1 positive subjects (P<0.005), and in smokers, compared to non-smokers. These effects are ascribed to the influence of GSTM1 genotype and of smoking status on DNA repair capacities. As the induction of CAs are associated with carcinogenesis, the challenge assay is able to detect enhanced susceptibility for CA caused by genetic predisposition of DNA repair deficiency. PMID- 11909762 TI - A new antimutagen from Mentha cordifolia Opiz. AB - The CHCl(3) extract of Mentha cordifolia Opiz. showed antimutagenicity against tetracycline. An antimutagen was purified by solvent partitioning and repeated normal phase-vacuum liquid chromatography (NP-VLC) using a micronucleus test guided isolation and purification. Spectral analyses showed that the isolated antimutagen is possibly 6,7-bis-(2,2-dimethoxyethene)-2,11-dimethoxy-2Z,4E,8E,10Z dodecatetraendioic acid. It inhibited the mutagenicity of tetracycline by 68.7% at a dosage of 0.01 mg per 20 g mouse. Statistical analysis using Kruskal-Wallis one-way analysis of variance (ANOVA) by ranks showed that its variance differs from that of the solvent control group (tetracycline+dimethylsulfoxide (DMSO)) at alpha=0.001. Moreover, the isolated antimutagen did not exhibit mutagenic activity at the same dosage. Statistical analysis showed that it is not mutagenic at 0.001 level of significance because its variance differs from that of tetracycline. PMID- 11909763 TI - Lymphocyte DNA damage in elevator manufacturing workers in Guangzhou, China. AB - AIMS: To study the effect of smoking, passive smoking, alcohol drinking, and occupational exposure to low level of benzene on DNA strand breaks in elevator manufacturing workers in Guangzhou, China. METHODS: Three hundred and fifty-nine workers (252 men and 107 women) of a modern elevator manufacturing factory, 205 were from production departments and 154 from managerial department. Information on the workers' health conditions, smoking, passive smoking, alcohol consumption and occupational exposure history was collected by personal interview. Lymphocyte DNA damage was measured by the Comet assay. RESULTS: None of the women smoked and 20.6% of the men were daily smokers. In non-smokers, the prevalence of passive smoking at work was 25% for men and 11.2% for women, and at home, 37.8 and 48.6%, respectively. Smoking significantly increased tail moment (P<0.001). Daily smokers had the largest tail moment (geometric mean, 95% CI) (0.93 microm (0.81 0.94)), followed by occasional smokers (0.76 microm (0.59-0.95)), ex-smokers (0.70 microm (0.58-0.85)), and never smokers (0.56 microm (0.53-0.60)). Tail moment increased significantly with daily tobacco consumption (cigarettes per day) (r=0.26, P<0.001) after adjusting for age, gender, occupational exposure, passive smoking, and drinking. Analysis of covariance (ANCOVA) showed that smoking (P<0.001), passive smoking at home (P=0.026), occupational exposure (P<0.001), male gender (P<0.001), and age (P=0.001) had independent effects on tail moment, whereas passive smoking at work and alcohol drinking had no significant effect. CONCLUSIONS: Smoking, passive smoking at home, male gender, age and occupational exposure independently increased lymphocyte DNA strand breaks. The presence of excess DNA damage under low level of occupational exposure to benzene or other solvents suggest that the current allowance concentrations may not be safe to prevent genotoxicity. PMID- 11909764 TI - Lead induced DNA strand breaks in lymphocytes of exposed workers: role of reactive oxygen species and protein kinase C. AB - Lead and lead compounds play a significant role in modern industry; a wide variety of population is at risk of occupational exposure and lead is suspected to be a human carcinogen. The biochemical and molecular mechanisms of lead toxicity are poorly understood, but emerging data suggest that some of the effects of lead may be due to its interference with calcium in the activation of protein kinase C (PKC) and/or through production of reactive oxygen species (ROS). Many of these results are conducted in vitro on cell lines or ex vivo on human lymphocytes treated in vitro. We, therefore, performed a study on the induction of DNA damage, using the alkaline comet assay, in lymphocytes of battery plant workers. To elucidate in vivo the mechanism(s) responsible for this effect, we determined ROS production, and glutathione (GSH) levels in living cells using the fluorescent probe (2',7'-dichlorofluorescein and monochlorobimane, respectively). Subcellular fractions were obtained from sonicated lymphocytes; cytosolic and membrane expression of PKC isoforms (alpha, and zeta) was evaluated after electrophoresis by immunoblot analysis. The results indicate that lead-exposed workers have significantly elevated levels of DNA breaks compared to the unexposed group. A multivariate analysis of variance (ANOVA) shows that the most common confounding factors (smoking, drinking and age) have no synergistic effects with lead-exposure on the comet parameters or on GSH levels and ROS production. The logistic regression analysis distinguishing the exposed and non-exposed indicates that only GSH with tail moment are selected as significant risk factors. There is a significant positive correlation with ROS production and negative correlation with GSH levels. The content of PKC alpha in cytosol and membranes is decreased 40% (indicating a down-regulation of protein), whereas PKC zeta isoform is not modified in an evident manner. Our results suggest that lead-exposure induces an increase of DNA breakage with an alternate cellular redox state and a significant down-regulation of PKC alpha, suggesting that this metal may act as a tumor promoter. PMID- 11909765 TI - A comparative study of genotoxic effects of anti-topoisomerase II drugs ICRF-193 and bufalin in Chinese hamster ovary cells. AB - With the ultimate purpose of testing the existence of possible differences in the effectiveness of the topoisomerase II catalytic inhibitor ICRF-193 (a bisdioxopiperazine) and the enzyme suppressor bufalin (a bufadienolide from toad venom) we have carried out a series of experiments aimed at inducing cytotoxicity as well as DNA and chromosome damage in transformed CHO cells. In order to assess any possible influence of DNA repair capacity of the treated cells on the final outcome, we have made use of the repair-defective CHO mutant EM9, which shows a defect in DNA single- and double-strand breaks repair for comparison with its repair-proficient parental line AA8. Our results seem to indicate that, while both ICRF-193 and bufalin suppress cell growth and result in a clear inhibition of topoisomerase II catalytic activity, only ICRF-193 has been shown as able to induce both chromosome and DNA damage, with a more pronounced effect in the CHO mutant EM9 than in the repair-proficient line AA8. PMID- 11909766 TI - Induction of sister chromatid exchanges and chromosome aberrations in cultured mammalian cells treated with aminophenylnorharman formed by norharman with aniline. AB - Aminophenylnorharman (APNH) is a newly identified mutagenic heterocyclic amine formed by coupling of norharman with aniline in the presence of S9 mix. Furthermore, mutagenic amino-3'-methylphenylnorharman (AMPNH) and aminophenylharman (APH) have been identified from a reaction mixture of norharman and o-toluidine and that of harman and aniline, respectively, with S9 mix. Among these three heterocyclic amines, APNH shows most potent mutagenic activity towards Salmonella typhimurium TA98 and YG1024 with S9 mix. In the present study, the induction of sister chromatid exchanges (SCEs) by APNH was examined in Chinese hamster lung (CHL) cells in vitro, comparing it to those of AMPNH and APH. On incubation with rat S9 for 6h, followed by a recovery culture period of 18h, a dose-dependent effect was found at concentrations between 0.00125 and 0.01 microg/ml for APNH and between 0.3125 and 5 microg/ml for AMPNH and APH. The approximate chemical concentrations leading to a three-fold of control SCE levels calculated from slopes of the linear regressions of induced SCEs were 0.005 for APNH, 0.51 for AMPNH and 1.7 microg/ml for APH. Because of the very strong SCE causing ability of APNH, we further explored its genotoxicity by examining the induction of chromosome aberrations in CHL cells. A dose-dependent effect was found for chromosome aberrations at concentrations between 0.00125 and 0.04 microg/ml of APNH. The aberrations observed were primarily chromatid exchanges (cte) and breaks (ctb). In conclusion, the potency of SCE induction and clastogenic activity induced by APNH is stronger than Actinomycin D, Mitomycin C (MMC) or 1,8-dinitropyrene which are considered to be the potent clastogens in the literature. Further studies are needed for elucidating mechanisms of the genotoxic actions of these compounds and for evaluating their potential hazards to human health. PMID- 11909767 TI - Mutagenic and carcinogenic heterocyclic amines as affected by muscle types/skin and cooking in pan-roasted mackerel. AB - The dependence on muscle types/skin and on degrees of cooking in the formation of heterocyclic amines (HCAs) of pan-roasted mackerel was studied. High levels of 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were found in very well done skin and ordinary muscle, being 4.2 and 5.3 ng/g, followed by 2-amino-3,4 dimethylimidazo[4,5-f] quinoxaline (MeIQx), being 1.8 and 2.1 ng/g and 2-amino-9H pyrido[2,3-b]indole (AalphaC), being 1.2 and 2.8 ng/g, respectively. In pan roasted mackerel, ordinary muscles contributed much more greatly to the formation of HCAs than skins due to its higher HCA contents and composition (76.1%). PMID- 11909768 TI - Antimutagenic potential of curcumin on chromosomal aberrations in Wistar rats. AB - Curcumin, a yellow pigment commonly used as a spice and food coloring agent is obtained from rhizomes of Curcuma longa and is a major chemopreventive component of turmeric. In the present set of investigations the antimutagenic potential of curcumin has been evaluated using in vivo chromosomal aberration assay in Wistar rats. Cyclophosphamide (CP), a well-known mutagen was given by intraperitoneal (i.p.) injection at the dose of 40 mg/kg body weight (b.w.). Curcumin was given at the dose of 100 and 200 mg/kg b.w. through gastric intubation for seven consecutive days prior to CP treatment. The animals were sacrificed at the sampling time of 24 h after treatment and their bone marrow tissue was analyzed for chromosomal damage and mitotic index. In CP treated animals a significant induction of chromosomal aberration was recorded with decrease in mitotic index. However, in curcumin-supplemented animals, no significant induction in chromosomal damage or change in mitotic index was recorded. In different curcumin supplemented groups, a dose dependent significant decrease in CP induced clastogenicity was recorded. The incidence of aberrant cells was found to be reduced by both the doses of curcumin when compared to CP treated group. The anticytotoxic potential of curcumin towards CP was also evident as the status of mitotic index was found to show increment. The study revealed the antigenotoxic potential of curcumin against CP induced chromosomal mutations. PMID- 11909769 TI - Preventing stroke. PMID- 11909770 TI - Africa can solve its own health problems. PMID- 11909771 TI - New treatments for varicose veins. PMID- 11909772 TI - Modern worries, new technology, and medicine. PMID- 11909773 TI - Air pollution and short term mortality. PMID- 11909774 TI - German doctors face investigation in drugs scandal. PMID- 11909775 TI - Woman at centre of Southall case faces prison sentence. PMID- 11909776 TI - Fewer new HIV infections in India. PMID- 11909777 TI - Intermediate care offers no "quick fix" solution. PMID- 11909778 TI - Mentally ill mother escapes death penalty but faces life imprisonment. PMID- 11909779 TI - Controversial immunologist faces court case. PMID- 11909780 TI - WHO insists screening can cut breast cancer rates. PMID- 11909781 TI - Ban on tobacco advertising moves a step closer. PMID- 11909782 TI - Influx of fake drugs to Nigeria worries health experts. PMID- 11909784 TI - Public sector must develop drugs for neglected diseases. PMID- 11909783 TI - WHO calls for safety of health workers in Palestinian territories. PMID- 11909785 TI - Use of ramipril in preventing stroke: double blind randomised trial. AB - OBJECTIVE: To determine the effect of the angiotensin converting enzyme inhibitor ramipril on the secondary prevention of stroke. DESIGN: Randomised controlled trial with 2x2 factorial design. SETTING: 267 hospitals in 19 countries. PARTICIPANTS: 9297 patients with vascular disease or diabetes plus an additional risk factor, followed for 4.5 years as part of the HOPE study. OUTCOME MEASURES: Stroke (confirmed by computed tomography or magnetic resonance imaging when available), transient ischaemic attack, and cognitive function. Blood pressure was recorded at entry to the study, after 2 years, and at the end of the study. RESULTS: Reduction in blood pressure was modest (3.8 mm Hg systolic and 2.8 mm Hg diastolic). The relative risk of any stroke was reduced by 32% (156 v 226) in the ramipril group compared with the placebo group, and the relative risk of fatal stroke was reduced by 61% (17 v 44). Benefits were consistent across baseline blood pressures, drugs used, and subgroups defined by the presence or absence of previous stroke, coronary artery disease, peripheral arterial disease, diabetes, or hypertension. Significantly fewer patients on ramipril had cognitive or functional impairment. CONCLUSION: Ramipril reduces the incidence of stroke in patients at high risk, despite a modest reduction in blood pressure. PMID- 11909786 TI - Relation between burden of disease and randomised evidence in sub-Saharan Africa: survey of research. AB - OBJECTIVE: To evaluate whether the amount of randomised clinical research on various medical conditions is related to the burden of disease and health needs of the local populations in sub-Saharan Africa. DESIGN: Construction and analysis of comprehensive database of randomised controlled trials in sub-Saharan Africa based on Medline, the Cochrane Controlled Trials Register, and several African databases. SETTING: Sub-Saharan Africa. MAIN OUTCOME MEASURES: Number of trials and randomised subjects for each category of disease in the global burden of disease taxonomy; ratios of disability adjusted life years (DALYs) per amount of randomised evidence. RESULTS: 1179 eligible randomised controlled trials were identified. The number of trials published each year increased over time. Almost half of the trials (n=565) had been done in South Africa. There was relatively good correlation between the estimated burden of disease at year 2000 and the number of trials performed (r=0.53, P=0.024) and the number of participants randomised (r=0.68, P=0.002). However,some conditions-for example, injuries (over 20 000 DALYs per patient ever randomised)-were more neglected than others. CONCLUSION: Despite recent improvements, few clinical trials are done in sub Saharan Africa. Clinical research in this part of the world should focus more evenly on the major contributors to burden of disease. PMID- 11909787 TI - Glycaemic control with continuous subcutaneous insulin infusion compared with intensive insulin injections in patients with type 1 diabetes: meta-analysis of randomised controlled trials. AB - OBJECTIVE: To compare glycaemic control and insulin dosage in people with type 1 diabetes treated by continuous subcutaneous insulin infusion (insulin infusion pump therapy) or optimised insulin injections. DESIGN: Meta-analysis of 12 randomised controlled trials. PARTICIPANTS: 301 people with type 1 diabetes allocated to insulin infusion and 299 allocated to insulin injections for between 2.5 and 24 months. MAIN OUTCOME MEASURES: Glycaemic control measured by mean blood glucose concentration and percentage of glycated haemoglobin. Total daily insulin dose. RESULTS: Mean blood glucose concentration was lower in people receiving continuous subcutaneous insulin infusion compared with those receiving insulin injections (standardised mean difference 0.56, 95% confidence interval 0.35 to 0.77), equivalent to a difference of 1.0 mmol/l. The percentage of glycated haemoglobin was also lower in people receiving insulin infusion (0.44, 0.20 to 0.69), equivalent to a difference of 0.51%. Blood glucose concentrations were less variable during insulin infusion. This improved control during insulin infusion was achieved with an average reduction of 14% in insulin dose (difference in total daily insulin dose 0.58, 0.34 to 0.83), equivalent to 7.58 units/day. CONCLUSIONS: Glycaemic control is better during continuous subcutaneous insulin infusion compared with optimised injection therapy, and less insulin is needed to achieve this level of strict control. The difference in control between the two methods is small but should reduce the risk of microvascular complications. PMID- 11909788 TI - National service framework for coronary heart disease: audit of English hospitals. PMID- 11909789 TI - Obstacles to answering doctors' questions about patient care with evidence: qualitative study. AB - OBJECTIVE: To describe the obstacles encountered when attempting to answer doctors' questions with evidence. DESIGN: Qualitative study. SETTING: General practices in Iowa. PARTICIPANTS: 9 academic generalist doctors, 14 family doctors, and 2 medical librarians. MAIN OUTCOME MEASURE: A taxonomy of obstacles encountered while searching for evidence based answers to doctors' questions. RESULTS: 59 obstacles were encountered and organised according to the five steps in asking and answering questions: recognise a gap in knowledge, formulate a question, search for relevant information, formulate an answer, and use the answer to direct patient care. Six obstacles were considered particularly salient by the investigators and practising doctors: the excessive time required to find information; difficulty modifying the original question, which was often vague and open to interpretation; difficulty selecting an optimal strategy to search for information; failure of a seemingly appropriate resource to cover the topic; uncertainty about how to know when all the relevant evidence has been found so that the search can stop; and inadequate synthesis of multiple bits of evidence into a clinically useful statement. CONCLUSIONS: Many obstacles are encountered when asking and answering questions about how to care for patients. Addressing these obstacles could lead to better patient care by improving clinically oriented information resources. PMID- 11909790 TI - Cancer chemoprevention. PMID- 11909791 TI - Broad complex tachycardia--Part I. PMID- 11909792 TI - Alteplase for stroke: money and optimistic claims buttress the "brain attack" campaign. PMID- 11909793 TI - Clinical problem solving and diagnostic decision making: selective review of the cognitive literature. PMID- 11909794 TI - MMR vaccine uptake may be lower than reported because of manipulation of target groups. PMID- 11909795 TI - MMR vaccine debate. Debate crystallises dilemma facing many medical disciplines. PMID- 11909796 TI - Physician assistants. Many general practitioners would welcome having physician assistants. PMID- 11909797 TI - European working time directive for doctors in training. Reduction in juniors' hours abolishes concept of continuity of care. PMID- 11909798 TI - Improving outcomes in depression. Integrated solutions should not be provided at the expense of reduced participation of statutory sector. PMID- 11909799 TI - The future of rehabilitation. Rehabilitation should be regarded as scientific challenge. PMID- 11909800 TI - Risky drinking by both sexes should be tackled. PMID- 11909801 TI - Time has come for mandatory unit labelling. PMID- 11909803 TI - Hospitals must not be closed without considering all factors. PMID- 11909802 TI - Child outpatient non-attendance may indicate welfare concerns. PMID- 11909804 TI - Death after inserting Hickman line was probably avoidable. PMID- 11909810 TI - Reduced transient outward K+ current and cardiac hypertrophy: causal relationship or epiphenomenon? PMID- 11909811 TI - Myosin light chain phosphatase: it gets around. PMID- 11909812 TI - Brain angiotensin II: new insights into its role in sympathetic regulation. PMID- 11909814 TI - Transcriptional regulation of vertebrate cardiac morphogenesis. AB - Transcription factors can regulate the expression of other genes in a tissue specific and quantitative manner and are thus major regulators of embryonic developmental processes. Several transcription factors that regulate cardiac genes specifically have been described, and the recent discovery that dominant inherited transcription factor mutations cause congenital heart defects in humans has brought direct medical relevance to the study of cardiac transcription factors in heart development. Although this field of study is extensive, several major gaps in our knowledge of the transcriptional control of heart development still exist. This review will concentrate on recent developments in the field of cardiac transcription factors and their roles in heart formation. PMID- 11909813 TI - CADASIL Notch3 mutant proteins localize to the cell surface and bind ligand. AB - Cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a vascular dementia arising from abnormal arteriolar vascular smooth muscle cells. CADASIL results from mutations in Notch3 that alter the number of cysteine residues in the extracellular epidermal growth factor-like repeats, important for ligand binding. It is not known whether CADASIL mutations lead to loss or gain of Notch3 receptor function. To examine the functional consequences of CADASIL mutations, we engineered 4 CADASIL-like mutations into rat Notch3 and have shown that the presence of an unpaired cysteine does not impair cell-surface expression or ligand binding. PMID- 11909815 TI - Matrix metalloproteinases: regulation and dysregulation in the failing heart. AB - Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes responsible for myocardial extracellular protein degradation. Several MMP species identified within the human myocardium may be dysregulated in congestive heart failure (CHF). For example, MMPs that are expressed at very low levels in normal myocardium, such as collagenase-3 (MMP-13) and the membrane-type-1 MMPs, are substantially upregulated in CHF. However, MMP species are not uniformly increased in patients with end-stage CHF, suggesting that a specific portfolio of MMPs are expressed in the failing myocardium. With the use of animal models of CHF, a mechanistic relationship has been demonstrated with respect to myocardial MMP expression and the left ventricular (LV) remodeling process. The tissue inhibitors of the MMPs (TIMPs) are locally synthesized proteins that bind to active MMPs and thereby regulate net proteolytic activity. However, there does not appear to be a concomitant increase in myocardial TIMPs during the LV remodeling process and progression to CHF. This disparity between MMP and TIMP levels favors a persistent MMP activation state within the myocardium and likely contributes to the LV remodeling process in the setting of developing CHF. The elucidation of upstream signaling mechanisms that contribute to the selective induction of MMP species within the myocardium as well as strategies to normalize the balance between MMPs and TIMPs may yield some therapeutic strategies by which to control myocardial extracellular remodeling and thereby slow the progression of the CHF process. PMID- 11909816 TI - Differential expression of adenosine receptors in human endothelial cells: role of A2B receptors in angiogenic factor regulation. AB - Adenosine has been reported to stimulate or inhibit the release of angiogenic factors depending on the cell type examined. To test the hypothesis that differential expression of adenosine receptor subtypes contributes to endothelial cell heterogeneity, we studied microvascular (HMEC-1) and umbilical vein (HUVEC) human endothelial cells. Based on mRNA level and stimulation of adenylate cyclase, we found that HUVECs preferentially express A2A adenosine receptors and HMEC-1 preferentially express A2B receptors. Neither cells expressed A1 or A3 receptors. The nonselective adenosine agonist 5'-N-ethylcarboxamidoadenosine (NECA) increased expression of interleukin-8 (IL-8), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) in HMEC-1, but had no effect in HUVECs. In contrast, the selective A2A agonist 2-p-(2 carboxyethyl)phenylethylamino-NECA (CGS 21680) had no effect on expression of these angiogenic factors. Cotransfection of each type of adenosine receptors with a luciferase reporter in HMEC-1 showed that A2B receptors, but not A1, A2A, or A3, activated IL-8 and VEGF promoters. These effects were mimicked by constitutively active alphaG(q), alphaG12, and alphaG13, but not alphaG(s) or alphaG(i1-3). Furthermore, stimulation of phospholipase C indicated coupling of A2B receptors to G(q) proteins in HMEC-1. Thus, differential expression of adenosine receptor subtypes contributes to functional heterogeneity of human endothelial cells. A2B receptors, predominantly expressed in human microvascular cells, modulate expression of angiogenic factors via coupling to G(q), and possibly via G12/13. PMID- 11909817 TI - Decreased nitric oxide synthesis in human endothelial cells cultured on type I collagen. AB - Endothelial dysfunction, considered as a defective vascular dilatation after certain stimuli, is characteristic of different pathological conditions, such as hypertension, atherosclerosis, or diabetes. A decreased synthesis or an increased degradation of nitric oxide (NO) has been postulated as the mechanism responsible for this alteration. The present experiments were designed to test the hypothesis that the presence of an abnormal extracellular matrix in vessel walls could be responsible for the decreased NO synthesis observed in these pathological conditions. Experiments were performed in cultured human umbilical vein endothelial cells (HUVECs) grown on type IV (Col. IV) or type I (Col. I) collagen. Cells seeded on Col. I showed decreased nitrite synthesis, nitric oxide synthase activity, eNOS protein content, and eNOS mRNA expression when compared with cells grown on Col. IV. Moreover, cells grown on Col. I failed to respond to glucose oxidase activation of the eNOS system. In both cases, the changes in the eNOS mRNA expression seemed to depend on the modulation of eNOS promoter activity. The downregulation of eNOS induced by Col. I was blocked by D6Y, a peptide that interferes with the Col. I-dependent signals through integrins, as well as by specific anti-integrin antibodies. Moreover, a decreased activation of integrin-linked kinase (ILK) may explain the effects observed in Col. I-cultured cells because the activity of this kinase was decreased in these cells and ILK modulation prevented the Col. I-induced changes in HUVECs. Taken together, these findings may contribute to explaining the basis of endothelial dysfunction in some vascular diseases. PMID- 11909818 TI - Differential association and localization of myosin phosphatase subunits during agonist-induced signal transduction in smooth muscle. AB - It has been known for some time that agonist-induced contractions of vascular smooth muscle are often associated with a sensitization of the contractile apparatus to intracellular Ca2+. One mechanism that has been suggested to explain Ca2+ sensitization is inhibition of myosin phosphatase activity. In the present study, we tested the hypothesis that differential localization of the phosphatase might be associated with its inhibition. Quantitative confocal microscopy of freshly dissociated, fully contractile smooth muscle cells was used in parallel with measurements of myosin light chain and myosin phosphatase phosphorylation. The results indicate that, in the smooth muscle cells, the catalytic and targeting subunits of the phosphatase are dissociated from each other in an agonist-specific manner and that the dissociation is accompanied by a slower rate of myosin phosphorylation. Targeting of myosin phosphatase to the cell membrane precedes the dissociation of subunits and is associated with phosphorylation of the targeting subunit at a Rho-associated kinase (ROK) phosphorylation site. The phosphorylation and membrane translocation of the targeting subunit are inhibited by a ROK inhibitor. This dissociation of subunits may provide a mechanism for the decreased phosphatase activity of phosphorylated myosin phosphatase. PMID- 11909819 TI - Intramolecular interaction of SUR2 subtypes for intracellular ADP-Induced differential control of K(ATP) channels. AB - ATP-sensitive K+ (K(ATP)) channels are composed of sulfonylurea receptors (SURs) and inwardly rectifying Kir6.2-channels. The C-terminal 42 amino acid residues (C42) of SURs are responsible for ADP-induced differential activation of K(ATP) channels in SUR-subtypes. By examining ADP-effect on K(ATP) channels containing various chimeras of SUR2A and SUR2B, we identified a segment of 7 residues at central portion of C42 critical for this phenomenon. A 3-D structure model of the region containing the second nucleotide-binding domain (NBD2) of SUR and C42 was developed based on the structure of HisP, a nucleotide-binding protein forming the bacterial Histidine transporter complex. In the model, the polar and charged residues in the critical segment located within a distance that allows their electrostatic interaction with Arg1344 at the Walker-A loop of NBD2. Therefore, the interaction might be involved in the control of ADP-induced differential activation of SUR2-subtype K(ATP) channels. PMID- 11909820 TI - Role of alpha4 integrin and VCAM-1 in CD18-independent neutrophil migration across mouse cardiac endothelium. AB - Myocardial damage due to reperfusion of ischemic tissue is caused primarily by infiltrating neutrophils. Although leukocyte beta2 integrins (CD18) play a critical role, significant neutrophil emigration persists when CD18 is neutralized or absent. This study examined the role of leukocyte beta1 integrin (alpha4) and its endothelial ligand VCAM-1 in CD18-independent neutrophil migration across cardiac endothelium. In a mouse model of myocardial ischemia and reperfusion, we show that compared with wild-type mice, neutrophil infiltration efficiency was reduced by 50% in CD18-null mice; in both types of mice, myocardial VCAM-1 staining increased after reperfusion. In wild-type mice, antibodies against CD18, ICAM-1 (an endothelial ligand for CD18), or VCAM-1 given 30 minutes before ischemia did not block neutrophil emigration at 3 hours reperfusion. Although anti-VCAM-1 attenuated neutrophil emigration by 90% in CD18 null mice, it did not diminish myocardial injury. To determine if CD18 independent neutrophil emigration was a tissue-specific response, we used isolated peripheral blood neutrophils from wild-type or CD18-null mice and showed neutrophil migration across lipopolysaccharide-activated cultured cardiac endothelium is CD18-independent, whereas migration across endothelium obtained from inferior vena cava is CD18-dependent. Consistent with our in vivo findings, migration of CD18-deficient neutrophils on cardiac endothelial monolayers is blocked by antibodies against alpha4 integrin or VCAM-1. We conclude tissue specific differences in endothelial cells account, at least partially, for CD18 independent neutrophil infiltration in the heart. PMID- 11909821 TI - Inhibition of nitrobenzylthioinosine-sensitive adenosine transport by elevated D glucose involves activation of P2Y2 purinoceptors in human umbilical vein endothelial cells. AB - Chronic incubation with elevated D-glucose reduces adenosine transport in endothelial cells. In this study, exposure of human umbilical vein endothelial cells to 25 mmol/L D-glucose or 100 micromol/L ATP, ATP-gamma-S, or UTP, but not ADP or alpha,beta-methylene ATP, reduced adenosine transport with no change in transport affinity. Inhibition of transport by D-glucose, ATP, and ATP-gamma-S was associated with reduced maximal binding, with no changes in the apparent dissociation constant for nitrobenzylthioinosine (NBMPR). A significant reduction (approximately 60+/-10%, P<0.05; n=6) in the number of human equilibrative NBMPR sensitive nucleoside transporters (hENT1s) per cell (1.8+/-0.1x10(6) in 5 mmol/L D-glucose) and in hENT1 mRNA levels was observed in cells exposed to D-glucose or ATP-gamma-S. Incubation with elevated D-glucose, but not with D-mannitol, increased the ATP release by 3+/-0.2-fold. The effects of D-glucose and nucleotides on the number and activity of hENT1 and hENT1 mRNA were blocked by reactive blue 2 (nonspecific P2Y purinoceptor antagonist), suramin (Galpha(s) protein inhibitor), or hexokinase but not by pyridoxal phosphate-6-azophenyl 2',4'-disulfonic acid (nonselective P2 purinoceptor antagonist). Our findings demonstrate that inhibition of adenosine transport via hENT1 in endothelial cells cultured in 25 mmol/L D-glucose could be due to stimulation of P2Y2 purinoceptors by ATP, which is released from these cells in response to D-glucose. This could be a mechanism to explain in part the vasodilatation observed in the early stages of diabetes mellitus or in response to D-glucose infusion. PMID- 11909822 TI - Reduction of I(to) causes hypertrophy in neonatal rat ventricular myocytes. AB - Prolonged action potential duration (APD) and decreased transient outward K+ current (I(to)) as a result of decreased expression of K(v4.2) and K(v4.3) genes are commonly observed in heart disease. We found that treatment of cultured neonatal rat ventricular myocytes with Heteropoda Toxin3, a blocker of cardiac I(to), induced hypertrophy as measured using cell membrane capacitance and (3)H leucine uptake. To dissect the role of specific I(to)-encoding genes in hypertrophy, I(to) was selectively reduced by overexpressing mutant dominant negative (DN) transgenes. I(to) amplitude was reduced equally (by about 50%) by overexpression of DN K(v1.4) (K(v1.4)N) or DN K(v4.2) (either K(v4.2)N or K(v4.2)W362F), but only DN K(v4.2) prolonged APD duration (at 1 Hz) and induced myocyte hypertrophy. This hypertrophy was prevented by coexpressing wild-type K(v4.2) channels (K(v4.2)F) with the DN K(v4.2) genes, suggesting the hypertrophy is due to I(to) reduction and not nonspecific effects of transgene overexpression. The hypertrophy caused by reductions of K(v4.x)-based I(to) was associated with increased activity of the calcium-dependent phosphatase, calcineurin, and could be prevented by coinfection with Ad-CAIN, a specific calcineurin inhibitor. The hypertrophy and calcineurin activation induced by K(v4.2)N infection were prevented by blocking Ca2+ entry and excitability with verapamil or high [K+]o. Our studies suggest that reductions of K(v4.2/3)-based I(to) play a role in hypertrophy signaling by activation of calcineurin. PMID- 11909823 TI - Role of heteromultimers in the generation of myocardial transient outward K+ currents. AB - Previous studies have demonstrated a role for Kv4 alpha subunits in the generation of the fast transient outward K+ current, I(to,f), in the mammalian myocardium. The experiments here were undertaken to explore the role of homomeric/heteromeric assembly of Kv4.2 and Kv4.3 and of the Kv channel accessory subunit, KChIP2, in the generation of mouse ventricular I(to,f). Western blots reveal that the expression of Kv4.2 parallels the regional heterogeneity in I(to,f) density, whereas Kv4.3 and KChIP2 are uniformly expressed in adult mouse ventricles. Antisense oligodeoxynucleotides (AsODNs) targeted against Kv4.2 or Kv4.3 selectively attenuate I(to,f) in mouse ventricular cells. Adenoviral mediated coexpression of Kv4.2 and Kv4.3 in HEK-293 cells and in mouse ventricular myocytes produces transient outward K+ currents with properties distinct from those produced on expression of Kv4.2 or Kv4.3 alone, and the gating properties of the heteromeric Kv4.2/Kv4.3 channels in ventricular cells are more similar to native I(to,f) than are the homomeric Kv4.2 or Kv4.3 channels. Biochemical studies reveal that Kv4.2, Kv4.3, and KChIP2 coimmunoprecipitate from adult mouse ventricles. In addition, most of the Kv4.2 and KChIP2 are associated with Kv4.3 in situ. Taken together, these results demonstrate that functional mouse ventricular I(to,f) channels are heteromeric, comprising Kv4.2/Kv4.3 alpha subunits and KChIP2. The results here also suggest that Kv4.2 is the primary determinant of the regional heterogeneity in I(to,f) expression in adult mouse ventricle. PMID- 11909824 TI - Hypertrophic cardiomyopathy in cardiac myosin binding protein-C knockout mice. AB - Familial hypertrophic cardiomyopathy (FHC) is an inherited autosomal dominant disease caused by mutations in sarcomeric proteins. Among these, mutations that affect myosin binding protein-C (MyBP-C), an abundant component of the thick filaments, account for 20% to 30% of all mutations linked to FHC. However, the mechanisms by which MyBP-C mutations cause disease and the function of MyBP-C are not well understood. Therefore, to assess deficits due to elimination of MyBP-C, we used gene targeting to produce a knockout mouse that lacks MyBP-C in the heart. Knockout mice were produced by deletion of exons 3 to 10 from the endogenous cardiac (c) MyBP-C gene in murine embryonic stem (ES) cells and subsequent breeding of chimeric founder mice to obtain mice heterozygous (+/-) and homozygous (-/-) for the knockout allele. Wild-type (+/+), cMyBP-C(+/-), and cMyBP-C(-/-) mice were born in accordance with Mendelian inheritance ratios, survived into adulthood, and were fertile. Western blot analyses confirmed that cMyBP-C was absent in hearts of homozygous knockout mice. Whereas cMyBP-C(+/-) mice were indistinguishable from wild-type littermates, cMyBP-C(-/-) mice exhibited significant cardiac hypertrophy. Cardiac function, assessed using 2 dimensionally guided M-mode echocardiography, showed significantly depressed indices of diastolic and systolic function only in cMyBP-C(-/-) mice. Ca2+ sensitivity of tension, measured in single skinned myocytes, was reduced in cMyBP C(-/-) but not cMyBP-C(+/-) mice. These results establish that cMyBP-C is not essential for cardiac development but that the absence of cMyBP-C results in profound cardiac hypertrophy and impaired contractile function. PMID- 11909825 TI - Inducible nitric oxide synthase modulates cyclooxygenase-2 activity in the heart of conscious rabbits during the late phase of ischemic preconditioning. AB - Cyclooxygenase-2 (COX-2) is known to mediate the cardioprotective effects of the late phase of ischemic preconditioning (PC); however, the signaling pathways involved in COX-2 induction following ischemic PC are unknown. In addition, although inducible nitric oxide synthase (iNOS) has been identified as a co mediator of late PC together with COX-2, the interaction between iNOS and COX-2 in the heart is unknown. Using conscious rabbits, we found that the induction of COX-2 expression 24 hours after ischemic PC was blocked by pretreatment with inhibitors of protein kinase C (PKC), Src protein tyrosine kinases (PTKs), and nuclear factor-kappaB (NF-kappaB) but not by inhibitors of NOS or scavengers of reactive oxygen species (ROS). The selective iNOS inhibitors SMT and 1400W, given 24 hours after PC, abrogated the increase in myocardial prostaglandin E2 (PGE2) and 6-keto-PGF1alpha, whereas the selective soluble guanylate cyclase inhibitor ODQ had no effect. COX-2 selective inhibitors (celecoxib and NS-398) did not affect iNOS activity. These results demonstrate that (i) ischemic PC upregulates cardiac COX-2 via PKC-, Src PTK-, and NF-kappaB-dependent signaling pathways, whereas generation of NO and ROS is not necessary, and (ii) the activity of newly synthesized COX-2 following PC requires iNOS-derived NO whereas iNOS activity is independent of COX-2-derived prostanoids, indicating that COX-2 is located downstream of iNOS in the protective pathway of late PC. The data also indicate that iNOS modulates COX-2 activity via cGMP-independent mechanisms. To our knowledge, this is the first demonstration that iNOS-derived NO drives prostanoid synthesis by COX-2 in the heart. NO-mediated activation of COX-2 may be a heretofore unrecognized mechanism by which NO exerts its salubrious effects in the late phase of PC. PMID- 11909826 TI - Inhibition of protein kinase Calpha prevents endothelial cell migration and vascular tube formation in vitro and myocardial neovascularization in vivo. AB - Although protein kinase C (PKC) activation is required for endothelial cell (EC) growth, migration, adhesion, and vessel formation, the role of individual PKC isoenzymes in these events is not defined. Because PKCalpha has been previously linked with enhanced EC migration and response to angiogenic growth factors, we characterized a specific phosphorothioate-modified 21-mer antisense PKCalpha (AS PKCalpha). AS-PKCalpha (500 nmol/L) prevented the expression of PKCalpha protein by 90% in human ECs and did not reduce the expression of any other PKC isoenzyme. AS-PKCalpha reduced human EC migration by 64% compared with its control oligonucleotide in a "scratch" wounding assay, and AS-PKCalpha reduced human EC adhesion to the extracellular matrix protein vitronectin by 18%. Phosphorylation of mitogen-activated protein kinase (extracellular signal-regulated kinase 1/2) induced by vascular endothelial growth factor was inhibited by 30% in human ECs transfected with AS-PKCalpha. Compared with control, AS-PKCalpha also reduced the number of EC tubes formed in a 3D type I collagen gel assay by 37.5%. Finally, using an osmotic minipump, we infused AS-PKCalpha into mice in which myocardial infarction was induced by coronary ligation and found that the oligonucleotide was primarily taken up by intramyocardial blood vessels. Compared with the results with control oligonucleotide, AS-PKCalpha oligonucleotide inhibited the number of anti-PKCalpha-stained blood vessels by 48% and reduced the total vessel number by 72% as well. In conclusion, the expression of PKCalpha is required for full EC migration, adhesion to vitronectin, vascular endothelial growth factor induced extracellular signal-regulated kinase activation, and tube formation and is likely to be of importance in myocardial angiogenesis in vivo after ischemia. PMID- 11909827 TI - Brain-selective overexpression of angiotensin (AT1) receptors causes enhanced cardiovascular sensitivity in transgenic mice. AB - To examine the physiological importance of brain angiotensin II type 1 (AT1) receptors, we developed a novel transgenic mouse model with rat AT1a receptors targeted selectively to neurons of the central nervous system (CNS). A transgene consisting of 2.8 kb of the rat neuron-specific enolase (NSE) 5' flanking region fused to a cDNA encoding the full open-reading frame of the rat AT1a receptor was constructed and transgenic mice (NSE-AT1a) were generated. Two of six transgenic founder lines exhibited brain-selective expression of the transgene at either moderate or high levels. Immunohistochemistry revealed widespread distribution of AT1 receptors in neurons throughout the CNS. This neuron-targeted overexpression of AT1a receptors resulted in enhanced cardiovascular responsiveness to intracerebroventricular (ICV) angiotensin II (Ang II) injection but not to other central pressor agents, demonstrating functional overexpression of the transgene in NSE-AT1a mice. Interestingly, baseline blood pressure (BP) was not elevated in either transgenic line. However, blockade of central AT1 receptors with ICV losartan caused significant falls in basal BP in NSE-AT1a mice but had no effect in nontransgenic controls. These results suggest that whereas there is an enhanced contribution of central AT1 receptors to the maintenance of baseline BP in NSE-AT1a mice, particularly effective baroreflex buffering prevents hypertension in this model. Used both independently, and in conjunction with mice harboring gene-targeted deletions of AT1a receptors, this new model will permit quantitative and relevant investigations of the role of central AT1a receptors in cardiovascular homeostasis in health and disease. PMID- 11909828 TI - Can we apply results from large to small arteries? PMID- 11909829 TI - Antimicrobial resistance: priorities for action. PMID- 11909830 TI - Guidelines for antibiotic usage in hospitals. PMID- 11909831 TI - Cardiotoxicity of fluoroquinolones. PMID- 11909832 TI - Mechanisms of antimicrobial action of antiseptics and disinfectants: an increasingly important area of investigation. PMID- 11909833 TI - Helicobacter pylori susceptibility testing by disc diffusion. AB - The bacterium Helicobacter pylori is found in c. 40% of the population and is responsible for the development of duodenal disease. Triple treatment with a proton-pump inhibitor or bismuth salt plus two antibiotics is now commonplace in all patients diagnosed. As antibiotic resistance reduces treatment efficacy, it is time to consider routine susceptibility testing to guide individual patient treatment and surveillance of antibiotic resistance. There are no published nationally agreed standards for disc diffusion testing of H. pylori. After reviewing the literature, we recommend the following method for disc diffusion tests. A suspension of cultures < or = 4 days old equivalent to McFarland Standard no. 4 (10(8) cfu/mL) should be used on Mueller-Hinton or Columbia agar base with 5-10% blood, using a metronidazole disc strength of 5 Ig and a clarithromycin disc strength of 2 microg. Anaerobic pre-incubation of plates is unnecessary. A H. pylori control susceptible to metronidazole (e.g. NCTC 12822) should be used. Zone sizes with the Mueller-Hinton agar base for metronidazole testing are <16 mm resistant, 16-21 mm intermediate and >21 mm susceptible. We suggest that isolates in the intermediate zone should be re-tested by Etest. Zone sizes with the Columbia agar base for metronidazole testing are <10 mm resistant and > or = 10 mm susceptible. Co-infection with two strains, which may be a mixture of isolates susceptible and resistant to metronidazole leading to conflicting susceptibility results, occurs in 5-10% of patients. Zone sizes with Mueller-Hinton agar and Columbia blood agar for clarithromycin testing are resistant no zone and susceptible any zone. PMID- 11909834 TI - Molecular characterization of ketolide-resistant erm(A)-carrying Staphylococcus aureus isolates selected in vitro by telithromycin, ABT-773, quinupristin and clindamycin. AB - The aim of this study was to investigate whether a Staphylococcus aureus strain that carried an inducibly expressed erm(A) gene might exhibit resistance to the non-inducers telithromycin, ABT-773, clindamycin, quinupristin, dalfopristin or the combination quinupristin-dalfopristin after incubation in the presence of inhibitory concentrations of any of these compounds. Whenever resistant mutants were obtained, these were investigated for the molecular basis of the altered resistance phenotype. Resistant mutants were not selected with dalfopristin or quinupristin-dalfopristin, but were obtained with the other four agents. Irrespective of which drug was used for selection, all mutants were cross resistant to clindamycin, quinupristin, telithromycin and ABT-773, and exhibited structural alterations in the erm(A) translational attenuator. The structural alterations observed included deletions of 14, 83, 121, 131, 147 or 157 bp, three different tandem duplications of 23, 25 or 26 bp, two different types of point mutation, as well as the insertion of IS256. All these alterations either completely prevented the formation of mRNA secondary structures in the erm(A) regulatory region or favoured the formation of those mRNA secondary structures that allowed translation of the erm(A) transcripts. Deletions, which were observed in almost two-thirds of the mutants, might be explained by illegitimate recombination between different parts of the erm(A) regulatory region. PMID- 11909835 TI - Identification of an immunodominant drug efflux pump in Burkholderia cepacia. AB - Burkholderia cepacia, a major pathogen amongst individuals with cystic fibrosis (CF), is intrinsically resistant to most clinically available antibiotics. We report the identification of an immunodominant antigen in CF patients infected with B. cepacia, a multidrug-resistance efflux pump called BcrA. The bcrA gene encodes a 46 kDa peptide with 14 potential alpha-helices that belongs to the major facilitator superfamily of drug transporters. A recombinant Escherichia coli strain was constructed containing the bcrA gene, which resulted in a four fold increase in resistance to tetracycline and an eight-fold increase in resistance to nalidixic acid. These results demonstrate that the bcrA gene is part of a drug efflux system that is potentially a major contributor to the high level antibiotic resistance observed in B. cepacia and thus a potential target for novel therapeutics. PMID- 11909836 TI - Increase in MICs of ciprofloxacin in vivo in two closely related clinical isolates of Enterobacter cloacae. AB - The mechanisms of fluoroquinolone resistance in two isolates of Enterobacter cloacae, Ecl#1 and Ecl#2, from the same patient and with identical pulsed-field gel electrophoresis patterns, have been analysed. MICs of ciprofloxacin were 0.25 and 1 mg/L for Ecl#1 and Ecl#2, respectively. Ecl#2 was also more resistant to chloramphenicol and organic solvents. The quinolone resistance determining regions of gyrA/B and parC/E, and the marORA and acrB genes, were sequenced. Expression of marR, acrB, soxS, robA, ramA and fis was analysed by northern blotting. The activity of a 90 bp E. cloacae mar promoter fragment was examined with the reporter plasmid pIGJ-1mar. Sequencing the gyrAB and parCE genes revealed a single amino acid substitution in GyrA (corresponding to position 83 in GyrA of Escherichia coli) in Ecl#1 and Ecl#2 (Phe83) compared with reference strain E. cloacae DSMZ 3264 (Thr83). Ecl#2 accumulated significantly less norfloxacin and displayed higher levels of expression of marR and acrB than Exl#1, indicative of greater fluoroquinolone efflux activity. Sequencing gyrB, parC/E and marORA, and northern blotting of robA, ramA and fis, did not reveal any further differences between the two strains. No homologue of soxRS was detected in E. cloacae. Expression of GFP from pIGJ1-mar in Ecl#2 was higher than in Ecl#1. In these two closely related clinical isolates of E. cloacae, a target mutation in GyrA (Ecl#1 and Ecl#2) and increased fluoroquinolone efflux by AcrAB (Ecl#2) contribute to the resistance phenotypes, corroborating findings in vitro and in vivo about the sequential development of fluoroquinolone resistance. PMID- 11909837 TI - Pseudomonas aeruginosa cells adapted to benzalkonium chloride show resistance to other membrane-active agents but not to clinically relevant antibiotics. AB - Our objective was to determine whether strains of Pseudomonas aeruginosa can adapt to growth in increasing concentrations of the disinfectant benzalkonium chloride (BKC), and whether co-resistance to clinically relevant antimicrobial agents occurs. Attempts were made to determine what phenotypic alterations accompanied resistance and whether these explained the mechanism of resistance. Strains were serially passaged in increasing concentrations of BKC in static nutrient broth cultures. Serotyping and genotyping were used to determine purity of the cultures. Two strains were examined for cross-resistance to other disinfectants and antibiotics by broth dilution MIC determination. Alterations in outer membrane proteins and lipopolysaccharide (LPS) expressed were examined by SDS-PAGE. Cell surface hydrophobicity and charge, uptake of disinfectant and proportion of specific fatty acid content of outer and cytoplasmic membranes were determined. Two P. aeruginosa strains showed a stable increase in resistance to BKC. Co-resistance to other quaternary ammonium compounds was observed in both strains; chloramphenicol and polymyxin B resistance were observed in one and a reduction in resistance to tobramycin observed in the other. However, no increased resistance to other biocides (chlorhexidine, triclosan, thymol) or antibiotics (ceftazidime, imipenem, ciprofloxacin, tobramycin) was detected. Characteristics accompanying resistance included alterations in outer membrane proteins, uptake of BKC, cell surface charge and hydrophobicity, and fatty acid content of the cytoplasmic membrane, although no evidence was found for alterations in LPS. Each of the two strains had different alterations in phenotype, indicating that such adaptation is unique to each strain of P. aeruginosa and does not result from a single mechanism shared by the whole species. PMID- 11909838 TI - In vitro action of carboxyfullerene. AB - Fullerene compounds have avid reactivity with free radicals and are regarded as 'radical sponges'. The trimalonic acid derivative of fullerene is one of the water-soluble compounds that has been synthesized and found to be an effective antioxidant both in vivo and in vitro. Carboxyfullerene has been shown to be effective in the treatment of both Gram-positive and -negative infections, although its mode of action is poorly understood. We determined the MIC and minimal bactericidal concentration of carboxyfullerene for 20 isolates, including Staphylococcus spp., Streptococcus spp., Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi and Klebsiella pneumoniae. We further investigated the action of carboxyfullerene using transmission electron microscopy (TEM), anticarboxyfullerene antibody binding assay and a membrane perturbation assay. All Gram-positive species were inhibited by < or = 50 mg/L of carboxyfullerene, whereas Gram-negative species were not inhibited, even at 500 mg/L carboxyfullerene. Bactericidal activity was demonstrated only for Gram positive species, particularly for Streptococcus pyogenes A-20, which was killed rapidly. Intercalation of carboxyfullerene into the cell wall of staphylococci and streptococci was demonstrated by TEM and anti-carboxyfullerene binding assay. Damage to the cell membrane in Gram-positive, but not Gram-negative, bacteria was confirmed by the membrane perturbation assay. These findings indicate that the action of carboxyfullerene on Gram-positive bacteria is achieved by insertion into the cell wall and destruction of membrane integrity. PMID- 11909839 TI - Evaluation of MIDITECH automated colorimetric MIC reading for antimicrobial susceptibility testing. AB - The MIDITECH colorimetric susceptibility test with automated reading is a modification of the standard broth microdilution method that uses a 3-(4,5 dimetylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) dye for detecting viable bacteria. The method can be applied to non-fastidious aerobic Gram negative bacteria, staphylococci and Enterococcus faecalis. To assess the reliability of this method, we compared susceptibility data obtained by this test with standard NCCLS microdilution assay results. For this purpose, 15 antibiotics and a well characterized set of 527 Gram-negative and Gram-positive bacterial isolates collected and stored at the Division of Infectious Diseases and Chemotherapy (Vienna General Hospital, Austria), yielding 5751 organism antibiotic combinations, were analysed in duplicate. The overall essential agreement (+/-1 log(2) dilution) between the MIDITECH and NCCLS methods was 96.18 +/- 0.67%. The colorimetric assay compared with the reference method produced MICs < or = 2 log(2) dilutions and > or = 2 log(2) dilutions in 2.34% and 1.48% comparisons, respectively. For 326 Gram-negative bacteria, the absolute interpretative agreement of both methods ranged from 87.12% for ampicillin sulbactam to 99.85% for meropenem (mean 94.86%); 417 (4.92%) minor, three (0.05%) major and 15 (0.63%) very major errors were found. For 127 staphylococci and 74 E. faecalis isolates, the absolute interpretative agreement ranged from 90.80% for ciprofloxacin to 100% for vancomycin and linezolid (mean 96.96%); 81 (2.77%) minor, three (0.15%) major and eight (0.83%) very major errors were found. For most of the clinically important aerobically growing pathogens, the MIDITECH colorimetric test provided reliable quantitative susceptibility data. The main advantage of this method is simple performance, automated reading and data processing without expensive investments. PMID- 11909840 TI - Antibiotic guidelines and antibiotic use in adult bacterial meningitis in The Netherlands. AB - In The Netherlands, national guidelines for the treatment of adult patients with bacterial meningitis were introduced in October 1997. In 1998 we began a prospective, nationwide study to evaluate the compliance with these consensus based guidelines. In addition, we evaluated whether the recommended initial treatment provides adequate microbiological coverage. From October 1998 to January 2000, 365 adults with bacterial meningitis were identified using information from The Netherlands Reference Laboratory for Bacterial Meningitis; 263 patients were classified into four categories depending on patient's age and underlying health status. In the first category, patients 16-60 years without risk factors, Neisseria meningitidis was the most common pathogen (53%); 62 of 127 patients (49%) received treatment in compliance with the guidelines. In the second and third categories, patients >60 years without risk factors and those with risk factors independently of age, Streptococcus pneumoniae caused 61% and 58% of cases, respectively. Compliance in these categories was about 17%. Overall, 33% of patients received treatment in compliance with the guidelines. The microbiological coverage of patients treated in compliance and not in compliance with the guidelines was 98% and 93%, respectively. In conclusion, 1 year after national consensus-based guidelines for the initial treatment of adult patient with bacterial meningitis were introduced in The Netherlands, only one third of Dutch physicians were adhering to the guidelines. The microbiological coverage for the patients who were treated in compliance with the guidelines was almost complete (98%). PMID- 11909841 TI - Antibiotic guidance on the web: an opportunity for open review. AB - The report The Path of Least Resistance recommended that local antibiotic prescribing information should be based on national guidelines produced under the aegis of the National Institute for Clinical Excellence (NICE). During the period when NICE guidelines on the management of infectious disease are being developed, interim guidelines are needed in primary care. Draft antibiotic guidance developed by Primary Care Groups and the Public Health Laboratory Service was posted on the PHLS website for consultation. An explanatory letter containing the website address was sent to all regional prescribing leads, asking them to draw it to the attention of all those involved in the development of antibiotic guidance. As a result of the numerous comments received from around the UK in the 18 months after posting on the website, 125 substantial changes were made and grading of evidence was added. The electronic production of guidance greatly facilitated the open review process and consequent modifications and reduced printing costs. PMID- 11909842 TI - In vitro bactericidal activity of ABT-773 and amoxicillin against erythromycin susceptible and -resistant strains of Streptococcus pyogenes. AB - The bactericidal activity of ABT-773 was compared with amoxicillin against 10 clinical isolates of S. pyogenes (six erythromycin susceptible and four erythromycin resistant). The MIC ranges (mg/L) were 0.004-0.25 of ABT-773 and 0.015-0.12 of amoxicillin. At 24 h, ABT-773 concentrations of 2 x MIC and 8 x MIC were bactericidal against three and six organisms, respectively. In comparison, amoxicillin was bactericidal against all 10 organisms at both test concentrations. PMID- 11909843 TI - Production of the RdxA protein in metronidazole-susceptible and -resistant isolates of Helicobacter pylori cultured from treated mice. AB - The objective of this study was to use immunoblotting with RdxA antisera to examine the production of the RdxA protein in mouse-derived metronidazole susceptible and -resistant isolates of Helicobacter pylori. A 24 kDa immunoreactive band corresponding to RdxA was observed in all 15 metronidazole susceptible and five of 50 metronidazole-resistant isolates. The rdxA gene of these five isolates contained missense mutations and transformation experiments confirmed that these mutations were associated with inactivation of the rdxA gene. No RdxA protein was produced in the other 45 metronidazole-resistant strains, including one in which the nucleotide sequence of the rdxA gene was unchanged. These results demonstrate a high correlation between production of the RdxA protein and susceptibility of H. pylori to metronidazole. Testing for the absence of the RdxA protein identifies the majority of strains that will respond poorly to metronidazole-containing eradication regimens. PMID- 11909845 TI - Alcoholic ingredients in skin disinfectants increase biofilm expression of Staphylococcus epidermidis. AB - The pathogenesis of Staphylococcus epidermidis is correlated with biofilm formation. We investigated the effect of three common alcoholic skin disinfectants, ethanol, n-propanol and isopropanol, on the biofilm formation of 37 clinical, icaADBC-positive S. epidermidis isolates. In alcohol-supplemented media 18 strains displayed increased biofilm expression. Sixteen of 19 strains were generally incapable of biofilm formation. In three representative isolates, the increase in biofilm formation was paralleled by increased polysaccharide intercellular adhesin synthesis. Regarding the widespread use of alcoholic skin disinfectants, it is possible that the alcohol-inducible biofilm phenotype of S. epidermidis could add to the development of foreign body-related infections. PMID- 11909844 TI - Comparison of phenotypic and genotypic methods for the detection of clarithromycin resistance in Mycobacterium avium. AB - The MICs of clarithromycin for 10 clinical isolates of Mycobacterium avium were determined using three methods: Bactec 460-TB, broth microdilution and Etest. The results were compared with the presence of resistance mutations in the 23S rRNA gene. Isolates were obtained from five AIDS patients who were treated with clarithromycin. Five isolates were recovered before and five during treatment. MICs were reproducible and comparable between the three methods. They were < or = 4 mg/L for pre-treatment isolates and > or = 128 mg/L for strains recovered during treatment. An MIC > or = 128 mg/L was associated with the presence of mutations in the 23S rRNA gene that were absent in the isolates exhibiting MIC < ro = 4 mg/L. PMID- 11909846 TI - Effect of amoxicillin and co-amoxiclav on the aerobic and anaerobic nasopharyngeal flora. AB - The effects of co-amoxiclav (AMC) and amoxicillin (AMX) therapy on the nasopharyngeal flora of children with acute otitis media (AOM) were compared. Nasopharyngeal culture for aerobic and anaerobic bacteria were obtained before therapy and 2-4 days after completion of antimicrobial therapy in 25 patients treated with either antibiotic. After therapy, 16 (64%) of the 25 patients treated with AMX and 23 (92%) of the 25 patients treated with AMC were considered clinically cured. Polymicrobial aerobic-anaerobic flora were present in all instances. A significant reduction in the number of both aerobic and anaerobic isolates occurred after therapy in those treated with AMX (177 isolates versus 133, P< 0.005) and AMC (172 isolates versus 60, P< 0.001). However, the number of all isolates recovered after therapy in those treated with AMC was significantly lower (60 isolates) than in those treated with AMX (133 isolates, P < 0.001). The recovery of known aerobic pathogens (e.g. Streptococcus pneumoniae, Staphylococcus aureus, beta-haemolytic streptococci, Haemophilus species and Moraxella catarrhalis) and penicillin-resistant bacteria after therapy was lower in the AMC group than in the AMX group (P < 0.005). This study illustrates the greater ability of AMC, compared with AMX, to reduce the number of potential nasopharyngeal pathogens and penicillin-resistant bacteria in children with AOM. PMID- 11909847 TI - An in vitro evaluation of the antibiotic/heparin lock to sterilize central venous haemodialysis catheters. AB - This in vitro study investigated the ability of antibiotic/heparin locks to sterilize central venous haemodialysis catheters (CVCs) inoculated with methicillin-resistant Staphylococcus epidermidis (MRSE). Isolates of MRSE were incubated in broth inside CVCs. The catheters were then drained and filled with either vancomycin/gentamicin/heparin (VGH), cefazolin/gentamicin/ heparin (CGH) or control locks for 48 h. The catheters were drained, filled with fresh broth and again incubated. The final catheter solutions were sampled and the remaining volumes filtered. The samples, filters and catheter segments were examined for growth. For two isolates, both the VGH and CGH locks sterilized the catheters. Bacterial counts of the remaining two isolates were significantly reduced by >99%, but the catheters were not sterilized after the instillation of a single antibiotic/heparin lock. PMID- 11909848 TI - Effect of an efflux pump inhibitor on the MIC of nalidixic acid for Acinetobacter baumannii and Stenotrophomonas maltophilia clinical isolates. PMID- 11909849 TI - In vitro activity of the novel des-F(6) quinolone BMS-284756 against genetically characterized clinical streptococcal isolates, including isolates with reduced quinolone susceptibility. PMID- 11909850 TI - In vitro antimicrobial susceptibility of clinical isolates of Aeromonas caviae, Aeromonas hydrophila and Aeromonas veronii biotype sobria. PMID- 11909851 TI - Catalytic properties of rice alpha-oxygenase. A comparison with mammalian prostaglandin H synthases. AB - Long-chain fatty acids can be metabolized to C(n)(-1) aldehydes by alpha oxidation in plants. The reaction mechanism of the enzyme has not been elucidated. In this study, a complete nucleotide sequence of fatty acid alpha oxygenase gene in rice plants (Oryza sativa) was isolated. The deduced amino acid sequence showed some similarity with those of mammalian prostaglandin H synthases (PGHSs). The gene was expressed in Escherichia coli and purified to apparently homogeneous state. It showed the highest activity with linoleic acid and predominantly formed 2-hydroperoxide of the fatty acid (C(n)), which is then spontaneously decarboxylated to form corresponding C(n)(-1) aldehyde. With linoleic or linoleic acids as a substrate, rice alpha-oxygenase formed no product having a lambda(max) at approximately 234 nm, which indicated that the enzyme could not oxygenize the pentadiene system in the substrate. The spectroscopic feature of the purified enzyme in its ferrous state is similar to that of mammalian PGHS, whereas that of dithionite-reduced state showed significant difference. Site-directed mutagenesis revealed that His-158, Tyr-380, and Ser-558 were essential for the alpha-oxygenase activity. These residues are conserved in PGHS and known as a heme ligand, a source of a radical species to initiate oxygenation reaction and a residue involved in substrate binding, respectively. This finding suggested that the initial step of the oxygenation reaction in alpha oxygenase has a high similarity with that of PGHS. The rice alpha-oxygenase activity was inhibited by imidazole but hardly inhibited by nonsteroidal anti inflammatory drugs, such as aspirin, ibuprofen, and flurbiprofen, which are known as typical PGHS inhibitors. In addition, peroxidase activity could not be detected with alpha-oxygenase when palmitic acid 2-hydroperoxide was used as a substrate. From these findings, the catalytic resemblance between alpha-oxygenase and PGHS seems to be evident, although there still are differences in their substrate recognitions and peroxidation activities. PMID- 11909852 TI - Isolation and characterization of a human STAT1 gene regulatory element. Inducibility by interferon (IFN) types I and II and role of IFN regulatory factor 1. AB - The transcription factor STAT1 plays a pivotal role in signal transduction of type I and II interferons (IFNs). STAT1 activation leads to changes in expression of key regulatory genes encoding caspases and cell cycle inhibitors. Deficient STAT1 expression in human cancer cells and virally mediated inhibition of STAT1 function have been associated with cellular resistance to IFNs and mycobacterial infection in humans. Thus, given the relative importance of STAT1, we isolated and characterized a human STAT1 intronic enhancer region displaying IFN-regulated activity. Functional analyses by transient expression identified a repressor region and type I and II IFN-inducible elements within the STAT1 enhancer sequence. A candidate IRF-E/GAS/IRF-E (IGI) sequence containing GAAANN nucleotide repeats was shown by gel shift assay to bind to IFN regulatory factor-1 (IRF-1), but not to IFN-stimulated gene factor-3 (ISGF-3) or STAT1-3. An additional larger IGI-binding complex containing IRF-1 was identified. Mutation of the GAAANN repeats within the IGI DNA element eliminated IRF-1 binding and the IFN-regulated activity of the STAT1 intronic enhancer region. Transfection of the IFN-resistant MM96 cell line to express increased levels of IRF-1 protein also elevated STAT1, STAT2, and p48/IRF-9 expression and enhanced cellular responsiveness to IFN-beta. Reciprocating regulation between IRF-1 and STAT1 genes and encoded proteins indicates that an intracellular amplifier circuit exists controlling cellular responsiveness to the IFNs. PMID- 11909853 TI - Regulation of TRAF2 signaling by self-induced degradation. AB - Receptors belonging to the tumor necrosis factor receptor (TNF-R) family utilize cytoplasmic adapter proteins called TNF-R-associated factors (TRAFs) as key elements in their signaling pathways. However, it is not yet clear how individual TRAFs regulate signaling by this large and growing receptor family. Signaling via the TNF-R family member CD40 has recently been shown to result in recruitment of TRAF2 to plasma membrane detergent-resistant microdomains (lipid rafts) as well as to subsequently initiate TRAF2 degradation. As TRAF2 associates with most members of the TNF-R family, we wished to determine how this degradation occurs. We show here that CD40-mediated TRAF2 degradation requires the zinc-binding RING domain of TRAF2 and is preceded by TRAF2 ubiquitination, suggesting that the TRAF2 RING may promote ubiquitination although the RING itself is not a target of ubiquitination. Several approaches show that ubiquitination and proteasomal activity are integral to TRAF2 degradation, and inhibition of this process potentiates CD40 signaling. PMID- 11909854 TI - A novel, extraneuronal role for cyclin-dependent protein kinase 5 (CDK5): modulation of cAMP-induced apoptosis in rat leukemia cells. AB - A number of cyclin-dependent protein kinase (CDK) inhibitors were tested for the ability to protect IPC-81 rat leukemic cells against cAMP-induced apoptosis. A near perfect proportionality was observed between inhibitor potency to protect against cAMP-induced apoptosis and to antagonize CDK5, and to a lesser extent, CDK2 and CDK1. Enforced expression of dominant negative CDK5 (but not CDK1-dn or CDK2-dn) protected against death, indicating that CDK5 activity was necessary for cAMP-induced apoptosis. The CDK inhibitors failed to protect the cells against daunorubicine-, staurosporine-, or okadaic acid-induced apoptosis. The inhibition of CDK5 prevented the cleavage of pro-caspase-3 in cAMP-treated cells. The cells could be saved closer to the moment of their onset of death by inhibitors of caspases than by inhibitors of CDK5. This suggested that the action of CDK5 was upstream of caspase activation. The cAMP treatment resulted in a moderate increase of the level of CDK5 mRNA and protein in IPC-81 wild-type cells. Such cAMP induction of CDK5 was not observed in cells expressing the inducible cAMP early repressor. The cAMP-induced increase of CDK5 contributed to apoptosis since cells overexpressing CDK5-wt were more sensitive for cAMP-induced death. These results demonstrate the first example of a proapoptotic CDK action upstream of caspase activation and of an extra-neuronal effect of CDK5. PMID- 11909855 TI - Purification and characterization of a cytosolic, 42-kDa and Ca2+-dependent phospholipase A2 from bovine red blood cells: its involvement in Ca2+-dependent release of arachidonic acid from mammalian red blood cells. AB - It has become evident that a Ca(2+)-dependent release of arachidonic acid (AA) and subsequent formation of bioactive lipid mediators such as prostaglandins and leukotrienes in red blood cells (RBCs) can modify physiological functions of neighboring RBCs and platelets. Here we identified a novel type of cytosolic PLA(2) in bovine and human RBCs and purified it to apparent homogeneity with a 14,000-fold purification. The purified enzyme, termed rPLA(2), has a molecular mass of 42 kDa and reveals biochemical properties similar to group IV cPLA(2), but shows different profiles from cPLA(2) in several column chromatographies. Moreover, rPLA(2) did not react with any of anti-cPLA(2) and anti-sPLA(2) antibodies and was identified as an unknown protein in matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. Divalent metal ions tested exhibited similar effects between rPLA(2) and cPLA(2), whereas mercurials inhibited cPLA(2) but had no effect on rPLA(2). Antibody against the 42-kDa protein not only precipitated the rPLA(2) activity, but also reacted with the 42-kDa protein from bovine and human RBCs in immunoblot analysis. The 42-kDa protein band was selectively detected in murine fetal liver cells known as a type of progenitor cells of RBCs. It was found that EA4, a derivative of quinone newly developed as an inhibitor for rPLA(2), inhibited a Ca(2+) ionophore-induced AA release from human and bovine RBCs, indicating that this enzyme is responsible for the Ca(2+)-dependent AA release from mammalian RBCs. Finally, erythroid progenitor cell assay utilizing diaminobenzidine staining of hemoglobinized fetal liver cells showed that rPLA(2) detectable in erythroid cells was down-regulated when differentiated to non-erythroid cells. Together, our results suggest that the 42-kDa rPLA(2) identified as a novel form of Ca(2+)-dependent PLA(2) may play an important role in hemostasis, thrombosis, and/or erythropoiesis through the Ca(2+)-dependent release of AA. PMID- 11909856 TI - Loss of plasma membrane phospholipid asymmetry requires raft integrity. Role of transient receptor potential channels and ERK pathway. AB - Cholesterol-rich membrane microdomains, also termed lipid rafts, are implicated in the recruitment of essential proteins for intracellular signal transduction. In nonstimulated cells, phosphatidylserine, an anionic aminophospholipid essential for the hemostatic response, is mostly sequestered in the inner leaflet of the plasma membrane. Cell stimulation by Ca(2+)-mobilizing or apoptogenic agents induces the migration of phosphatidylserine to the exoplasmic leaflet, allowing the assembly and activation of several key enzyme complexes of the coagulation cascade and phagocyte recognition of stimulated or senescent cells. We have recently proposed that store-operated Ca(2+) entry regulates externalization of phosphatidylserine at the cell surface (Kunzelmann-Marche, C., Freyssinet, J.-M., and Martinez, M. C. (2001) J. Biol. Chem. 276, 5134-5139). Here, we show that store-operated Ca(2+) entry and phosphatidylserine exposure are dramatically reduced after raft disruption by methyl-beta-cyclodextrin. In addition, transient receptor potential channel 1-specific antibody was able to significantly decrease Ca(2+)-induced redistribution of phosphatidylserine. Furthermore, store-operated Ca(2+) entry and phosphatidylserine exposure were dependent in part on the extracellular signal-regulated kinase pathway associated with rafts. Hence, raft integrity and store-operated Ca(2+) entry involving transient receptor potential channel 1 channels are essential for completion of the phosphatidylserine transmembrane redistribution process. PMID- 11909857 TI - Metastable macromolecular complexes containing high mobility group nucleosome binding chromosomal proteins in HeLa nuclei. AB - High mobility group nucleosome-binding (HMGN) proteins belong to a family of nuclear proteins that bind to nucleosomes and enhance transcription from chromatin templates by altering the structure of the chromatin fiber. The intranuclear organization of these proteins is dynamic and related to the metabolic state of the cell. Here we report that approximately 50% of the HMGN proteins are organized into macromolecular complexes in a fashion that is similar to that of other nuclear activities that modify the structure of the chromatin fiber. We identify several distinct HMGN-containing complexes that are relatively unstable and find that the inclusion of HMGN in the complexes varies according to the metabolic state of the cell. The nucleosome binding ability of HMGN in the complex is stronger than that of the free HMGN. We suggest that the inclusion of HMGN proteins into metastable multiprotein complexes serves to target the HMGN proteins to specific sites in chromatin and enhances their interaction with nucleosomes. PMID- 11909858 TI - Mutational study on the roles of disulfide bonds in the beta-subunit of gastric H+,K+-ATPase. AB - The gastric proton pump, H(+),K(+)-ATPase, consists of the catalytic alpha subunit and the non-catalytic beta-subunit. Correct assembly between the alpha- and beta-subunits is essential for the functional expression of H(+),K(+)-ATPase. The beta-subunit contains nine conserved cysteine residues; two are in the cytoplasmic domain, one in the transmembrane domain, and six in the ectodomain. The six cysteine residues in the ectodomain form three disulfide bonds. In this study, we replaced each of the cysteine residues of the beta-subunit with serine individually and in several combinations. The mutant beta-subunits were co expressed with the alpha-subunit in human embryonic kidney 293 cells, and the role of each cysteine residue or disulfide bond in the alpha/beta assembly, stability, and cell surface delivery of the alpha- and beta-subunits and H(+),K(+)-ATPase activity was studied. Mutant beta-subunits with a replacement of the cytoplasmic and transmembrane cysteines preserved H(+),K(+)-ATPase activity. All the mutant beta-subunits with replacement(s) of the extracellular cysteines did not assemble with the alpha-subunit, resulting in loss of H(+),K(+)-ATPase activity. These mutants did not permit delivery of the alpha-subunit to the cell surface. Therefore, each of these disulfide bonds of the beta-subunit is essential for assembly with the alpha-subunit and expression of H(+),K(+)-ATPase activity as well as for cell surface delivery of the alpha-subunit. PMID- 11909859 TI - Analysis of heterophilic and homophilic interactions of cadherins using the c Jun/c-Fos dimerization domains. AB - Cadherin-mediated cell-cell adhesion is initiated by cis dimerization of cadherin ectodomains at the cell surface followed by an antiparallel trans interaction of dimers on opposing cells. To resolve open questions concerning the molecular details and specificity of cis and trans interactions, ectodomains of E- and P cadherin were analyzed by chemical cross-linking and by electron microscopy. At the high intrinsic concentration created by artificial oligomerization the N terminal cadherin (CAD)-domain of P-cadherin are forming ring-like cis dimers. At 2 mm Ca(2+)-associated rings involving two cis dimers indicate trans contacts in electron micrographs. cis and trans interactions were further analyzed by heterodimerization of the ectodomains of E-cadherin (ECAD) and P-cadherin (PCAD) through the leucine zipper domains of c-Jun and c-Fos. ECADJun/ECADFos dimers predominantly form ring-like cis dimers at 1 mm Ca(2+) and double-ringed trans contacts above 2 mm Ca(2+). The Ca(2+)-dependent tetrameric trans contacts of ECADJun/ECADFos dimers are also detectable after chemical cross-linking. Only cis contacts but no trans interactions are observed for heterodimers of ECADFos and the Trp-2 to Ala mutant ECADW2AJun arguing for a decisive role of Trp-2 in trans but not cis interaction. Neither cis nor trans interaction was found for heterodimers of ECADJun and PCADFos suggesting that specificity for homophilic interactions already exists at the level of cis dimerization. PMID- 11909860 TI - The viral transactivator E1A regulates the mouse mammary tumor virus promoter in an isoform- and chromatin-specific manner. AB - Proteins encoded by the adenovirus E1A gene regulate both cellular and viral genes to mediate effects on cell cycle, differentiation, and cell growth control. We have identified the mouse mammary tumor virus (MMTV) promoter as a target of E1A action and investigated the role nucleoprotein structure plays in its response to E1A. Both 12 and 13 S forms target the MMTV promoter when it has a disorganized and accessible chromatin configuration. However, whereas the 13 S form is stimulatory, the 12 S form is repressive. When the MMTV promoter adopts an organized and repressed chromatin structure, it is targeted only by the 13 S form, which stimulates it. Although evidence indicates that E1A interacts with the SWI/SNF remodeling complex, E1A had no effect on chromatin remodeling at the MMTV promoter in organized chromatin. Analysis of E1A mutants showed that stimulation of the MMTV promoter is mediated solely through conserved region 3 and does not require interaction with Rb, p300/CBP-associated factor, or CBP/p300. Imaging analysis showed that E1A colocalizes with MMTV sequences in vivo, suggesting that it functions directly at the promoter. These results indicate that E1A stimulates the MMTV promoter in a fashion independent of chromatin conformation and through a direct mechanism involving interaction with the basal transcription machinery. PMID- 11909861 TI - Cell death and mechanoprotection by filamin a in connective tissues after challenge by applied tensile forces. AB - Cells in mechanically challenged environments must cope with high amplitude forces to maintain cell viability and tissue homeostasis. Currently, force induced cell death and the identity of mechanoprotective factors are not defined. We examined death in cultured periodontal fibroblasts, connective tissue cells that are exposed to heavy applied forces in vivo. Static tensile forces (0.48 piconewtons/microm2 cell area) were applied through magnetite beads coated with collagen or bovine serum albumin. There was a time-dependent increase of the percentage of propidium iodide-permeable cells in force-loaded cultures incubated with collagen but not bovine serum albumin beads, indicating a role for integrins. Cells exhibited reduced mitochondrial membrane potential, increased caspase-3 activation, nuclear condensation, terminal deoxynucleotidyl transferase nick end labeling staining, and detachment from the culture dish. The caspase-3 inhibitor acetyl-Asp-Glu-Val-Asp-aldehyde reduced detachment 3-fold. There was a rapid (<10-s) decrease in plasma membrane potential after force application, which, in filamin A-deficient melanoma cells, contributed to irreversible cell depolarization. In fibroblast cultures, cells with increased permeability to propidium iodide exhibited approximately 2-fold less filamin A content than impermeable cells. Fibroblasts transfected with antisense filamin A constructs or with filamin A constructs without an actin-binding domain exhibited 2-3-fold increased proportions of dead cells relative to controls. We conclude that high amplitude forces delivered through integrins can promote apoptosis in a proportion of cells and that filamin A confers mechanoprotection by preventing membrane depolarization. PMID- 11909862 TI - The unique centromeric chromatin structure of Schizosaccharomyces pombe is maintained during meiosis. AB - In meiosis I sister centromeres are unified in their polarity on the spindle, and this unique behavior is known to require the function of meiosis-specific factors that set some intrinsic property of the centromeres. The fission yeast, Schizosaccharomyces pombe, possesses complex centromeres consisting of repetitive DNA elements, making it an excellent model in which to study the behavior of complex centromeres. In mitosis, during which sister centromeres mediate chromosome segregation by establishing bipolar chromosome attachments to the spindle, the central core of the S. pombe centromere chromatin has a unique irregular nucleosome pattern. Deletion of repeats flanking this core structure have no effect on mitotic chromosome segregation, but have profound effects during meiosis. While this demonstrates that the outer repeats are critical for normal meiotic sister centromere behavior, exactly how they function and how monopolarity is established remains unclear. In this study we provide the first analysis of the chromatin structure of a complex centromere during meiosis. We show that the nature and extent of the unique central core chromatin structure is maintained with no measurable expansion. This demonstrates that monopolarity of sister centromeres, and subsequent reversion to bipolarity, does not involve a global change to the centromeric chromatin structure. PMID- 11909863 TI - Isolation and identification of the major heparan sulfate proteoglycans in the developing bovine rib growth plate. AB - Heparan sulfate proteoglycans are thought to mediate the action of growth factors. The heparan sulfate-containing proteoglycans in extracts of the bovine fetal rib growth plate were detected using the monoclonal antibody 3G10, which recognizes a neoepitope generated by heparitinase digestion (David, G., Bai, X. M., Van der Schueren, B., Cassiman, J. J., and Van den Berghe, H. (1992) J. Cell Biol. 119, 961-975). The heparan sulfate proteoglycans that react with this antibody were identified using antisera to known proteoglycans; purified using CsCl density gradient centrifugation, molecular sieve, and ion exchange chromatography; and then characterized. The major heparan sulfate proteoglycans in the growth plate had core proteins of 200 kDa and larger and were identified as perlecan and aggrecan. These two heparan sulfate proteoglycans could be effectively separated from each other by CsCl density gradient centrifugation alone. Perlecan contained 25% heparan sulfate and 75% chondroitin sulfate. The heparan sulfate chains on growth plate perlecan were considerably smaller than the chondroitin sulfate chains, and the heparan sulfate disaccharide content was different than that found for heparan sulfate from either kidney, tumor tissue, or growth plate aggrecan. Aggrecan contained only 0.1% heparan sulfate, which was localized to the CS-1 domain of aggrecan. These results indicate that perlecan and aggrecan would be the principal candidate proteoglycans involved in the action of heparan sulfate-binding proteins in the developing growth plate. PMID- 11909864 TI - The protein phosphatase-1 regulator NIPP1 is also a splicing factor involved in a late step of spliceosome assembly. AB - NIPP1 is a ubiquitous regulator of protein phosphatase-1 (PP1) and is targeted to the splicing factor storage sites (speckles) in the nucleus by its forkhead associated domain. We show here that NIPP1 is also a component of the spliceosomes in HeLa cell-splicing extracts and that the interaction with the spliceosomes requires a functional forkhead-associated domain. The in vitro splicing of beta-globin pre-mRNA was not affected by exogenous wild type NIPP1 but was blocked by mutants that lacked residues 225-329. The inhibition by these dominant negative mutants resulted from a block in a late phase of spliceosome assembly, i.e. at the transition between the B-complex and the C-complex. Site directed mutagenesis furthermore showed that this spliceosomal function of NIPP1 was unrelated to its ability to bind PP1 or RNA. Our data suggest that NIPP1 can function independently as a splicing factor and a phosphatase regulator. PMID- 11909865 TI - Myc target in myeloid cells-1, a novel c-Myc target, recapitulates multiple c-Myc phenotypes. AB - Using cDNA microarrays, we recently identified a large number of transcripts that are regulated differentially by the c-Myc oncoprotein in myeloid cells. Here, we characterize one of these, termed MT-MC1 (Myc Target in Myeloid Cells-1). MT-MC1 is a widely expressed nuclear protein whose overexpression, unlike that of c-Myc targets reported previously, recapitulates multiple c-Myc phenotypes. These include promotion of apoptosis, alteration of morphology, enhancement of anchorage-independent growth, tumorigenic conversion, promotion of genomic instability, and inhibition of hematopoietic differentiation. The MT-MC1 promoter is a direct c-Myc target; it contains two consensus E-box elements, both of which bind c-Myc.Max heterodimers. Mutation of either site abrogates DNA binding by c Myc.Max and renders the promoter c-Myc unresponsive. Finally, MT-MC1 regulates the expression of several other c-Myc target genes. MT-MC1 represents a proximal and direct c-Myc target that recapitulates many of the properties typically associated with Myc oncoprotein overexpression. PMID- 11909866 TI - The major conformational IgE-binding epitopes of hevein (Hev b6.02) are identified by a novel chimera-based allergen epitope mapping strategy. AB - A novel approach to localize and reconstruct conformational IgE-binding epitope regions of hevein (Hev b6.02), a major natural rubber latex allergen, is described. An antimicrobial protein (AMP) from the amaranth Amaranthus caudatus was used as an immunologically non-IgE-binding adaptor molecule to which terminal or central parts of hevein were fused. Hevein and AMP share a structurally identical core region but have different N-terminal and C-terminal regions. Only 1 of 16 hevein-allergic patients showed weak IgE binding to purified native or recombinant AMP. Chimeric AMP with the hevein N terminus was recognized by IgE from 14 (88%) patients, and chimeric AMP with the hevein C terminus was recognized by IgE from 6 (38%) patients. In contrast, chimeric AMP containing the hevein core region was recognized by IgE from only two patients. When both the N terminal and C-terminal regions of hevein were fused with the AMP core, IgE from all 16 patients bound to the chimera. This chimera was also able to significantly inhibit (>70%) IgE binding to the native hevein. On the contrary, linear synthetic peptides corresponding to hevein regions in the AMP chimeras showed no significant IgE binding capacity in either enzyme-linked immunosorbent assay or inhibition enzyme-linked immunosorbent assay. These results suggest that the IgE binding ability of hevein is essentially determined by its N-terminal and C terminal regions and that major IgE-binding epitopes of hevein are conformational. The chimera-based epitope mapping strategy described here provides a valuable tool for defining structural epitopes and creating specific reagents for allergen immunotherapy. PMID- 11909868 TI - CX3CR1 tyrosine sulfation enhances fractalkine-induced cell adhesion. AB - Fractalkine is a unique CX(3)C chemokine/mucin hybrid molecule that functions like selectins in inducing the capture of receptor-expressing cells. Because of the importance of tyrosine sulfation for ligand binding of the selectin ligand PSGL1, we tested the role of tyrosine sulfation for CX(3)CR1 function in cell adhesion. Tyrosine residues 14 and 22 in the N terminus of CX(3)CR1 were mutated to phenylalanine and stably expressed on K562 cells. Cells expressing CX(3)CR1 Y14F were competent in signal transduction but defective in capture by and firm adhesion to immobilized fractalkine under physiologic flow conditions. In static binding assays, CX(3)CR1-Y14F mutants had a 2-4-fold decreased affinity to fractalkine compared with wild type CX(3)CR1. By surface plasmon resonance measurements of fractalkine binding to biosensor chip-immobilized cell membranes, CX(3)CR1-Y14F mutants had a 100-fold decreased affinity to fractalkine. CX(3)CR1 expressing cell membranes treated with arylsulfatase to desulfate tyrosine residues also showed a 100-fold decreased affinity for fractalkine. Finally, synthesized, sulfated N-terminal CX(3)CR1 peptides immobilized on biosensor chips showed a higher affinity for fractalkine than non-sulfated peptides. Thus, we conclude that sulfation of tyrosine 14 enhances the function of CX(3)CR1 in cell capture and firm adhesion. Further, tyrosine sulfation may represent a general mechanism utilized by molecules that function in the rapid capture of circulating leukocytes. PMID- 11909867 TI - Sphingomyelin modulates the transbilayer distribution of galactosylceramide in phospholipid membranes. AB - The interrelationships among sphingolipid structure, membrane curvature, and glycosphingolipid transmembrane distribution remain poorly defined despite the emerging importance of sphingolipids in curved regions and vesicle buds of biomembranes. Here, we describe a novel approach to investigate the transmembrane distribution of galactosylceramide in phospholipid small unilamellar vesicles by (13)C NMR spectroscopy. Quantitation of the transbilayer distribution of [6 (13)C]galactosylceramide (99.8% isotopic enrichment) was achieved by exposure of vesicles to the paramagnetic ion, Mn(2+). The data show that [6 (13)C]galactosylceramide prefers (70%) the inner leaflet of phosphatidylcholine vesicles. Increasing the sphingomyelin content of the 1-palmitoyl-2-oleoyl phosphatidylcholine vesicles shifted galactosylceramide from the inner to the outer leaflet. The amount of galactosylceramide localized in the inner leaflet decreased from 70% in pure 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles to only 40% in 1-palmitoyl-2-oleoyl-phosphatidylcholine/sphingomyelin (1:2) vesicles. The present study demonstrates that sphingomyelin can dramatically alter the transbilayer distribution of a monohexosylceramide, such as galactosylceramide, in 1-palmitoyl-2-oleoyl-phosphatidylcholine/sphingomyelin vesicles. The results suggest that sphingolipid-sphingolipid interactions that occur even in the absence of cholesterol play a role in controlling the transmembrane distributions of cerebrosides. PMID- 11909869 TI - Species-specific inhibition of porphobilinogen synthase by 4-oxosebacic acid. AB - Porphobilinogen synthase (PBGS) catalyzes the condensation of two molecules of 5 aminolevulinic acid (ALA), an essential step in tetrapyrrole biosynthesis. 4 Oxosebacic acid (4-OSA) and 4,7-dioxosebacic acid (4,7-DOSA) are bisubstrate reaction intermediate analogs for PBGS. We show that 4-OSA is an active site directed irreversible inhibitor for Escherichia coli PBGS, whereas human, pea, Pseudomonas aeruginosa, and Bradyrhizobium japonicum PBGS are insensitive to inhibition by 4-OSA. Some variants of human PBGS (engineered to resemble E. coli PBGS) have increased sensitivity to inactivation by 4-OSA, suggesting a structural basis for the specificity. The specificity of 4-OSA as a PBGS inhibitor is significantly narrower than that of 4,7-DOSA. Comparison of the crystal structures for E. coli PBGS inactivated by 4-OSA versus 4,7-DOSA shows significant variation in the half of the inhibitor that mimics the second substrate molecule (A-side ALA). Compensatory changes occur in the structure of the active site lid, which suggests that similar changes normally occur to accommodate numerous hybridization changes that must occur at C3 of A-side ALA during the PBGS-catalyzed reaction. A comparison of these with other PBGS structures identifies highly conserved active site water molecules, which are isolated from bulk solvent and implicated as proton acceptors in the PBGS catalyzed reaction. PMID- 11909870 TI - Juvenile hormone diol kinase. II. Sequencing, cloning, and molecular modeling of juvenile hormone-selective diol kinase from Manduca sexta. AB - The gene sequence of Manduca sexta juvenile hormone diol kinase (JHDK) codes for an enzyme that has 59% sequence identity to Drosophila melanogaster sarcoplasmic calcium-binding protein-2 (dSCP2). JHDK and dSCP2 are similar to G-proteins with three conserved sequence elements involved in purine nucleotide binding. Both proteins contain two pairs of EF-hand motifs. Characterization and partial purification of the D. melanogaster homolog of M. sexta JHDK from adult D. melanogaster gave material with JHDK activity. This activity has an experimental pI and molecular mass that are nearly identical to those of dSCP2. Moreover, D. melanogaster phosphotransferase activity has very similar chromatographic retention in three systems compared with M. sexta JHDK. Substrate docking to three-dimensional models of JHDK has shown that the three conserved nucleotide binding elements surround the putative substrate-binding site and align with conserved sequence elements of p21(Ras) and adenylate kinase. D. melanogaster dSCP2 is a homolog of M. sexta JHDK, and these proteins constitute a novel kinase family that binds nucleotides using the scaffold of an SCP (Protein Data Bank code ). PMID- 11909871 TI - Juvenile hormone diol kinase. I. Purification, characterization, and substrate specificity of juvenile hormone-selective diol kinase from Manduca sexta. AB - Manduca sexta juvenile hormone diol kinase (JHDK) catalyzes the conversion of juvenile hormone (JH) diol to JH diol phosphate. JHDK may be the first example of a phosphotransferase directly involved in the catabolism and inactivation of a lipid-soluble hormone. JHDK is an enzyme crucial for secondary metabolism of JH and possesses high specificity and catalytic efficiency for JH diol. In this study, the purification and characterization of native JHDK are described; its enzymatic properties are examined; and its role in cellular JH metabolism is explored. Using a variety of potential substrates, we show that JHDK has a preference for ATP, but will catalyze the formation of JH diol phosphate with GTP as the phosphate donor. JHDK has a nanomolar K(m) for JH I diol and a low micromolar value for MgATP. JH II and III diols also serve as phosphate acceptors with low micromolar K(m), whereas other diol derivatives of terpenoid esters structurally similar to JH metabolites are not phosphorylated. The reaction proceeds via a sequential Bi Bi mechanism. JHDK is active as a homodimer with a subunit molecular mass of 20 kDa. JHDK binds 5'-p-fluorosulfonylbenzoyladenosine and is inhibited by micromolar levels of Ca2+. PMID- 11909872 TI - Bax-mediated Ca2+ mobilization promotes cytochrome c release during apoptosis. AB - Previous studies have demonstrated that Ca(2+) is released from the endoplasmic reticulum (ER) in some models of apoptosis, but the mechanisms involved and the functional significance remain obscure. We confirmed that apoptosis induced by some (but not all) proapoptotic stimuli was associated with caspase-independent, BCL-2-sensitive emptying of the ER Ca(2+) pool in human PC-3 prostate cancer cells. This mobilization of ER Ca(2+) was associated with a concomitant increase in mitochondrial Ca(2+) levels, and neither ER Ca(2+) mobilization nor mitochondrial Ca(2+) uptake occurred in Bax-null DU-145 cells. Importantly, restoration of DU-145 Bax expression via adenoviral gene transfer restored ER Ca(2+) release and mitochondrial Ca(2+) uptake and dramatically accelerated the kinetics of staurosporine-induced cytochrome c release, demonstrating a requirement for Bax expression in this model system. In addition, an inhibitor of the mitochondrial Ca(2+) uniporter (RU-360) attenuated mitochondrial Ca(2+) uptake, cytochrome c release, and DNA fragmentation, directly implicating the mitochondrial Ca(2+) changes in cell death. Together, our data demonstrate that Bax-mediated alterations in ER and mitochondrial Ca(2+) levels serve as important upstream signals for cytochrome c release in some examples of apoptosis. PMID- 11909873 TI - Identification of peptaibols from Trichoderma virens and cloning of a peptaibol synthetase. AB - The fungus Trichoderma virens is a ubiquitous soil saprophyte that has been applied as a biological control agent to protect plants from fungal pathogens. One mechanism of biocontrol is mycoparasitism, and T. virens produces antifungal compounds to assist in killing its fungal targets. Peptide synthetases produce a wide variety of peptide secondary metabolites in bacteria and fungi. Many of these are known to possess antibiotic activities. Peptaibols form a class of antibiotics known for their high alpha-aminoisobutyric acid content and their synthesis as a mixture of isoforms ranging from 7 to 20 amino acids in length. Here we report preliminary characterization of a 62.8-kb continuous open reading frame encoding a peptaibol synthetase from T. virens. The predicted protein structure consists of 18 peptide synthetase modules with additional modifying domains at the N- and C-termini. T. virens was shown to produce a mixture of peptaibols, with the largest peptides being 18 residues. Mutation of the gene eliminated production of all peptaibol isoforms. Identification of the gene responsible for peptaibol production will facilitate studies of the structure and function of peptaibol antibiotics and their contribution to biocontrol activity. PMID- 11909874 TI - HIV Tat binds Egr proteins and enhances Egr-dependent transactivation of the Fas ligand promoter. AB - HIV Tat can enhance activation-induced up-regulation of Fas ligand (FasL), which may contribute to T cell apoptosis in human immune deficiency virus (HIV) infected individuals. We have assessed functional and physical interactions between Tat and the Egr family of transcription factors (Egr-1, -2, and -3), the latter two of which are major participants in activation-induced FasL up regulation. Here we report that whereas Tat itself has no effect on the FasL promoter, it binds to Egr-2 and -3 and synergizes with them to superinduce expression of a FasL promoter-driven reporter. A Tat molecule containing a single amino acid substitution that results in the loss of transactivation activity for the HIV long terminal repeat still binds Egr-3 but can no longer enhance Egr mediated transactivation of the FasL promoter. Furthermore, the mutated Tat acts as a dominant negative inhibitor, blocking the superinduction of FasL caused by wild type Tat. Because Tat is present in virus-infected cells and in the serum of HIV-infected individuals, these results suggest that increased expression of FasL in these circumstances may result from the cooperative activities of activation induced Egrs and Tat. PMID- 11909875 TI - Coordination of ATF6-mediated transcription and ATF6 degradation by a domain that is shared with the viral transcription factor, VP16. AB - ATF6 is a 670-amino acid endoplasmic reticulum (ER) transmembrane protein that is cleaved in response to ER stress. The resulting N-terminal fragment of approximately 400 amino acids translocates to the nucleus and activates selected ER stress-inducible genes, such as GRP-78 and sarco/endoplasmic reticulum ATPase, which are required for cell survival. In studying the mechanism of ATF6-activated transcription, we found that when HeLa cells were transfected with a plasmid encoding ATF6-(1-373), ER stress-inducible reporter gene activation was high, but ATF6-(1-373) expression was low, unless a proteasome inhibitor was added. In contrast, transfection with a plasmid encoding ATF6-(94-373) resulted in low reporter activation and high expression of ATF6-(94-373), which was independent of the proteasome inhibitor. Thus, the information responsible for transcriptional activation and proteasomal degradation must lie within the N terminal 93 amino acids of ATF6. This portion of ATF6 was found to be homologous to the herpes simplex viral protein, VP16. One 8-amino acid domain of particular interest in this region of ATF6 is 75% identical to the VN8 region in VP16. VN8 is required for VP16-mediated transcription, as well as rapid degradation of VP16 by proteasomes. Point mutations in the VN8-like region of ATF6 caused a loss of transcription, increased expression levels, and an increase in half-life. Thus, the potent transcriptional activities and rapid degradation of ATF6 and VP16 require the VN8 domains in each protein. Homology searches indicate that ATF6 is the only eukaryotic protein known that possesses an active VN8 domain, raising questions about how this domain evolved and the functional importance underlying its appearance in only these two transcription factors. PMID- 11909876 TI - A case-only approach for assessing gene by sex interaction in human longevity. AB - As one aspect of the complex feature of longevity, gene by sex interaction plays an important role in influencing human life span. With advances in molecular genetics, more studies aimed at assessing gene by sex interaction are expected. New and valid statistical methods are needed. In this article, we introduce a nontraditional approach, the case-only design, which was originally proposed for assessing gene and disease associations, to detect gene by sex interaction in human longevity. Applications of this method to data collected from centenarian studies show that it can produce consistent results as compared with results obtained from case-control and other approaches. The method cannot be used as a substitute for traditional case-control studies since it is limited to the detection of interactions only. However, the easily applicable case-only approach can be an important tool for screening many potential genes that contribute to human longevity. PMID- 11909877 TI - Effect of L-carnitine on brain lipid peroxidation and antioxidant enzymes in old rats. AB - The effect of L-carnitine on lipid peroxidation and enzymatic antioxidants, such as superoxide dismutase, catalase, and glutathione peroxidase, was evaluated in brain regions of young and old rats. In all brain regions except the hypothalamus, lipid peroxidation was higher for old rats than for young control rats. The activity of superoxide dismutase, glutathione peroxidase, and catalase was lower in the striatum, cerebral cortex, and hippocampus, but no difference was observed in the hypothalamus and cerebellum. L-Carnitine administration (intraperitoneally) prevented thiobarbituric acid-reactive substance formation in the cerebral cortex, cerebellum, hypothalamus, hippocampus, and striatum of 24 month-old rats. Administration of L-carnitine reversed the age-associated changes in a duration-dependent manner. Results suggest that the neuroprotective effect on the brains in old rats was achieved by the elevation of antioxidants with L carnitine. PMID- 11909879 TI - Functionally relevant thresholds of quadriceps femoris strength. AB - The purpose of this study was to identify quadriceps femoris strength thresholds below which performance on ambulatory tasks is compromised. A second purpose was to evaluate whether self-reported functioning matches evaluated performance of the activities. Subjects (N = 100; age 73 +/- 0.9 years) participated in isometric knee extension strength tests, performed three functional ambulatory tasks (chair rise, gait speed, and stair ascent and descent), and answered standard survey questions assessing physical function. Significant relationships were observed between functional performance and the ratio of isometric leg extension peak torque to body weight (STR/WT) for each activity (p <.0001). For each activity, thresholds of STR/WT between 3.0 and 3.5 N m/kg were observed, below which the likelihood for success was reduced. Thresholds were determined by calculating the value of STR/WT that minimized the classification error. Individuals with a STR/WT < 3.0 N m/kg are at a substantial risk for impaired function in chair rise, gait speed, and stair ascent and descent. Sensitivity and specificity of STR/WT as a predictor of functional success ranged from 76% to 81% and from 78% to 94%, respectively, depending on activity. This is of clinical significance, as the STR/WT thresholds can identify individuals with preclinical disability (beginning to have difficulty with ambulatory tasks) as opposed to those in whom an outright disability is observed. This may be useful for targeting individuals for strengthening interventions and developing specific intervention goals. PMID- 11909878 TI - Maintenance of whole muscle strength and size following resistance training in older men. AB - Following a progressive resistance training (PRT) program of 3 days per week, we sought to examine how effective a resistance training maintenance program of 1 day per week would be to preserve muscle strength and size in older men. Each subject's whole muscle strength (1 repetition maximum, or 1RM) and whole muscle size (determined by computed tomography scan) were measured before (T1) and after (T2) 12 weeks of PRT and again following 6 months (T3) of training (TR) or detraining (DT). During the 12-week PRT, older men (N = 10; age 70 +/- 4 years) trained their knee extensors 3 days per week at 80% of their 1RM. The maintenance program consisted of older men (n = 5; TR; 75 +/- 1 years) who completed 3 sets of 10 repetitions at 80% of their 1RM 1 day per week (this was equivalent to a single training session that was performed 3 days per week during the 12-week PRT). The other group of older men (n = 5; DT; 69 +/- 1 years) resumed their normal lifestyle (no regular physical activity) following the 12-week PRT. From T1 to T2, muscle strength increased (p <.05) 45% (66 +/- 10 to 94 +/- 10 kg) in the TR group and 53% (50 +/- 6 to 74 +/- 7 kg) in the DT group. From T2 to T3, whole muscle strength of the TR group was unchanged (96 +/- 11 kg), whereas strength decreased (p <.05) in the DT group by 11% (66 +/- 6 kg). Muscle size demonstrated a similar pattern with a 7% increase (p <.05) in both groups from T1 to T2. No change in muscle size was found in the TR group from T2 to T3, whereas the DT group had a 5% reduction (p <.05). These data indicate that resistance training 1 day per week was sufficient to maintain muscle strength and size in these older men following a 12-week PRT program. Furthermore, the men who resumed their normal lifestyle (no regular physical activity) experienced significant losses in muscle strength and size. PMID- 11909880 TI - Effect of calorie restriction on mortality kinetics in inbred strains of mice following 7,12-dimethylbenz[a]anthracene treatment. AB - Calorie restriction (CR) has long been known to increase longevity and to delay the onset and to decrease the incidence of many age-related disease processes. The mechanism(s) by which these outcomes are attained is unidentified. This experiment was designed to examine whether differences existed in the extent to which various inbred strains of mice respond to CR. This work explored whether carcinogen-treated animals could be used to facilitate this aim by decreasing the time needed to observe differences in mortality kinetics between CR mice and ad libitum (AL) fed controls. Female mice from each of eight strains (A/J, BALB/c, C3H, C57BL/6, DBA/2J, FVB/J, NMRI, and 129/J) were given a single oral dose (65 mg/kg) of the carcinogen 7,12-dimethylbenz[a]anthracene and subsequently fed AL or calorically restricted. Following carcinogen treatment, the spectrum of lesions observed demonstrated genotypic variability, thereby complicating comparison among the inbred strains examined. However, in terms of the magnitude of alteration in mortality kinetics observed, a statistical analysis revealed that differences exist among the various strains of mice in their response. PMID- 11909882 TI - Avoidance and accommodation of surface height changes by healthy, community dwelling, young, and elderly men. AB - The purpose of this study was to compare avoidance and accommodation strategies during gait between healthy, community-dwelling, young and elderly males. Ten young (28.4 +/- 5.4 years) and ten elderly (69.5 +/- 6.1 years) males were tested while walking on the level, avoiding a 11.75-cm-high obstacle, and accommodating a change in floor height of 11.75 cm. Three dimensional, kinematic, and kinetic analyses were performed bilaterally. Both age groups used the same general sagittal plane strategies in order to avoid and accommodate the obstructions, but the elderly group had significantly reduced lead toe clearances. These riskier toe clearances by the elderly group were found to be due to limited, frontal plane pelvic motion, shorter stride lengths, and subtle differences in the interplay of lower limb energetic patterns within the sagittal plane. These results are discussed with respect to diminishing physical capacity in the elderly populations. PMID- 11909881 TI - Hormonal responses to endurance and resistance exercise in females aged 19-69 years. AB - Thirty cross-trained, female subjects (19-69 years) completed an endurance exercise session (ES), a resistance exercise session (RS), and a control session (CS) in a randomized, balanced design. The ES consisted of 40 minutes of cycling at 75% maximum heart rate, and the RS consisted of 3 sets of 10 repetitions of eight exercises. During the CS, subjects performed no exercise. Before and after exercise, and after 30 minutes of recovery, blood samples were analyzed for plasma lactate and serum growth hormone, insulin-like growth factor 1, testosterone, estradiol, dehydroepiandrosterone, and cortisol. Samples were taken during the CS at the same intervals as during the exercise sessions. There were no age-related differences in intensity measures during exercise. Absolute change from baseline in testosterone (p <.001), estradiol (p <.05), and growth hormone (p <.01) was significantly greater in the ES and RS compared with that in the CS. Absolute change in dehydroepiandrosterone was significantly greater in the RS only (p <.05). Results indicate that an acute bout of exercise can increase concentrations of anabolic hormones in females across a wide age range. PMID- 11909883 TI - Qualitative research: what does it have to offer to the gerontologist? AB - The growing challenges of promoting health and managing illness in an ever changing health care system require an arsenal of research approaches. Qualitative methods have a long tradition in disciplines such as sociology and anthropology and are being used with greater frequency as interdisciplinary health-related disciplines attempt to understand and explain complex problems. The purpose of this article is to define and describe the main features of qualitative research and to examine ways in which this methodology is relevant and useful in gerontological studies. A concise comparison of quantitative and qualitative methods is made, and suggestions are provided for when qualitative approaches are useful. A review of the three most common approaches is provided. Most important, references are provided for those gerontologists interested in learning more about qualitative research methods. PMID- 11909884 TI - Maintenance of BMD in older male runners is independent of changes in training volume or VO(2)peak. AB - BACKGROUND: The results of several recent research studies have questioned the benefit of chronic running in reducing the risk of developing osteoporosis. These negative conclusions are based upon the findings that runners, in general, are not different from controls with regard to their bone mineral density (BMD). This has led one to speculate that the value of weight-bearing exercise might be related to its ability to maintain bone mass or, perhaps, decrease the expected rates of loss. METHODS: We examined a group of 54 male master athletes ranging in age from 40 to 80 years longitudinally over a 5- to 7-year intervening period. Physiological parameters included VO(2)peak, body composition, bone density, and running performance. Medical histories and training records were obtained via questionnaire. RESULTS: Over the average 4.6 years between tests one and two, significant increases in body weight and lean body mass were observed. Aerobic fitness declined, as did weekly mileage and 5K and 10K performance times. Bone mineral density was lower in the whole body but not in the hip or the spine. Finally, we report no significant relationship between the change in training volume, change in body weight or lean mass, and change in aerobic capacity with changes in BMD. CONCLUSIONS: Hip and spine BMD are maintained over a 4- to 5-year period in master runners. Furthermore, changes in bone density or content are not sensitive to moderate changes in training volumes. We conclude that bone density can be maintained by running in older active men. These findings suggest that if a minimal threshold of mileage is required, the level is certainly below the average mileage of master runners. PMID- 11909885 TI - Late life function and disability instrument: I. Development and evaluation of the disability component. AB - BACKGROUND: Efforts to evaluate the effectiveness of clinical and community-based interventions designed to impact late-life disability have been hindered significantly by limitations in current instrumentation. More conceptually sound and responsive measures of disability are needed. METHODS: Applying Nagi's disablement model, we wrote questionnaire items that assessed disability in terms of frequency and limitation in performance of 25 life tasks. We evaluated their validity and test-retest reliability with 150 ethnically and racially diverse adults aged 60 and older who had a range of functional limitations, using factor analysis and Rasch analytic techniques to examine and refine the instrument. RESULTS: Our analyses resulted in a 16-item disability component with two dimensions, one focused on frequency of performance and the other addressing limitation in performance of life tasks, with two disability domains within each dimension. The frequency dimension consisted of a personal and a social role domain, and the limitation dimension consisted of an instrumental and a management role domain. Expected differences in summary scores of known functional limitation groups support the validity of this instrument. Test-retest intraclass correlations of the reproducibility of each overall dimension summary score were moderate to high (intraclass correlation coefficients .68-.82). CONCLUSIONS: The Late-Life Function and Disability Instrument has potential to assess meaningful concepts of disability across a wide variety of life tasks with relatively few items. PMID- 11909887 TI - Complementary and alternative medicine (CAM) use by older adults: a comparison of self-report and physician chart documentation. AB - BACKGROUND: Older adults are increasingly using complementary and alternative medicine (CAM) dietary supplements and herbal remedies, but may not discuss this with their physicians. When patients do report using CAM, their physicians may not record this information in patient charts. METHODS: This cross-sectional analysis compared results of a convenience sample survey with medical charts. Participants were older than 65 and from an urban academic hospital's ambulatory geriatrics practice. We measured (i) prevalence of CAM use; (ii) proportion of CAM supplements and herbs (CAMsh) reported by patients and documented in patients' charts; (iii) percentage of patients reporting taking CAMsh with anticoagulant activity (ginger, ginkgo, garlic, and vitamin E) while concomitantly taking prescribed anticoagulant medications, as per chart; and (iv) percentages of those patients for whom the CAM anticoagulant was or was not documented in the chart. RESULTS: We surveyed 212 patients; of those, 182 had available charts. Prevalence of CAM use was 64%. Only 35% of all self-reported supplements were documented in the charts. Of 182 patients, 84 (46%) reported taking CAM with anticoagulant properties: of these, 52% took a prescribed anticoagulant (per chart), while 48% took CAM but not prescribed anticoagulants. CONCLUSION: CAM use is highly prevalent among older adults. Physicians do not consistently record the use of CAMsh on patients' charts. This may lead to unrecognized, potentially harmful drug-herb/drug-supplement interactions. Physicians should elicit and document information on CAM use from older adult patients, both to provide sound medical care and to advance knowledge about drug herb/drug-supplement interactions. PMID- 11909886 TI - Late Life Function and Disability Instrument: II. Development and evaluation of the function component. AB - BACKGROUND: Self-reported capability in physical functioning has long been considered an important focus of research for older persons. Current measures have been criticized, however, for conceptual confusion, lack of sensitivity to change, poor reproducibility, and inability to capture a wide range of upper and lower extremity functioning. METHODS: Using Nagi's disablement model, we wrote physical functioning questionnaire items that assessed difficulty in 48 common daily tasks. We constructed the instrument using factor analysis and Rasch analytic techniques and evaluated its validity and test-retest reliability with 150 ethnically and racially diverse adults aged 60 years and older who had a range of functional limitations. RESULTS: Our analyses resulted in a 32-item function component with three dimensions--upper extremity, basic lower extremity, and advanced lower extremity functions. Expected differences in summary scores of known-functional limitation groups support its validity. Test-retest stability over a 1- to 3-week period was extremely high (intraclass correlation coefficients =.91 to.98). CONCLUSIONS: The Late-Life Function and Disability Instrument has potential to assess activity concepts related to upper and lower extremity functioning across a wide variety of daily physical tasks and individual levels of physical functioning. PMID- 11909888 TI - The relationship between cognitive and physical performance: MacArthur Studies of Successful Aging. AB - BACKGROUND: The relationship between change in cognitive and physical performance has yet to be fully understood. Because aging decreases the ability to learn new information while preserving more established knowledge, this article examines whether the association between change in cognitive and physical performance depends on the nature of the physical task. METHODS: Data from the MacArthur Research Network on Successful Aging Community Study--a longitudinal three-site, cohort study of high-functioning, disability-free Americans aged 70 to 79 in 1988 (reinterviewed in 1991 and 1995)--are used for this investigation. We examine the association between change in cognitive performance and two categories of physical performance: novel/attentional demanding physical tasks (e.g., standing on a single leg) or routine physical tasks (e.g., walking at a normal pace). Change in physical performance (over 7 years) is regressed on change in cognitive performance (over the same period) controlling for baseline cognitive ability, demographic factors, health status, and behavioral characteristics. RESULTS: The findings suggest that declines in cognitive performance are associated with declines in both novel/attentional demanding and routine physical tasks. In addition to decline in cognition, gender, prevalent health conditions (e.g., cancer, high blood pressure, and the fracture of a hip), and smoking behavior are associated with decline in performance on some physical tasks. CONCLUSIONS: The findings suggest that the successful execution of physical tasks demands cognitive processes regardless of the nature of the task. Researchers using performance-based measures of physical functioning should pay particular attention to the cognitive capacities of their subjects and how these might influence their assessment. PMID- 11909889 TI - Sleep problems in a very old population: drug use and clinical correlates. AB - BACKGROUND: Complaints of disturbed or dissatisfied sleep are common among older people. This study aimed to evaluate the prevalence of sleep problems in very old persons and its relation to physical and mental health and drug use. METHODS: This is a cross-sectional analysis of sleep problems in a population of old persons living in Stockholm, Sweden. There were 641 subjects aged 81+ years; 77.8% were women, 91.4% were noninstitutionalized, and 68.6% lived alone. All persons underwent a comprehensive medical and psychiatric interview and examination. Sleep problems were assessed using the Clinical Psychopathological Rating Scale (CPRS). Covariates included chronic medical conditions, depression, dementia, pain, self-rated health, activities of daily living, use of hypnotics sedatives, use of other psychotropic drugs, and use of nonpsychotropic drugs. RESULTS: More than one third of subjects were identified with sleep problems. They were more common among women and persons using a higher number of drugs. Poor self-rated health, depression, and pain were related to the presence of sleep problems. Among persons with sleep problems and depression, only 19.2% used antidepressants, and 46.2% used hypnotics-sedatives. Among persons with sleep problems and pain, 63.2% used analgesics, and 47.0% used hypnotics-sedatives. One or more chronic diseases, use of hypnotics-sedatives, use of other psychotropic drugs, and use of nonpsychotropic drugs were also related to sleep problems. After multivariate analysis, factors significantly related to sleep problems were female gender, depression, pain, and hypnotic-sedative use. CONCLUSIONS: Sleep problems were common in this very old population. These results suggest the importance of carefully assessing an older person's complaints to accurately diagnose and effectively treat sleep problems. PMID- 11909891 TI - Effects of age, step direction, and reaction condition on the ability to step quickly. AB - BACKGROUND: The ability to take a step quickly is important for balance maintenance during activities of daily living. The purpose of this study was to investigate the effects of age, reaction condition, and step direction on the ability to take a volitional step as fast as possible. METHODS: The performance of a voluntary step task was measured in young adult (mean age 20, SD 0.9 years), young-old adult (mean age 67, SD 3.7 years), and old adult (mean age 78, SD 2.3 years) healthy female participants. Each participant stepped as fast as possible in eight directions in response to a visual cue in a simple or choice reaction time condition. The effects of age, reaction condition, and step direction and their interactions on the primary outcome variables of response time, step liftoff, and step landing time were examined. RESULTS: The normal aging process progressively increased the response, liftoff, and landing times. The choice reaction time condition, compared to the simple, had significantly increased response, liftoff, and landing times. Step direction significantly affected the liftoff and landing times, with lateral, diagonal, and anterior and posterioir (A P) times increasing, respectively. CONCLUSIONS: We found substantial declines in the ability to step rapidly in healthy adults as age increased. When a decision was required regarding the step direction, the step performance also declined. Step direction also significantly affected step performance. The assessment of voluntary step performance, which may be an indicator of balance ability, should include dimensions of both direction and the choice condition. PMID- 11909892 TI - Consumer responses to the Wisconsin Partnership Program for Elderly Persons: a variation on the PACE Model. AB - BACKGROUND: The Wisconsin Partnership Program (WPP) is a variation on the Program for All-inclusive Care of the Elderly (PACE) model that is designed to be more flexible by allowing frail elderly dual-eligible (for both Medicare and Medicaid) clients to use their regular primary care physicians instead of relying on the physician hired by PACE. Case management is provided by a team of nurse, social worker, and nurse practitioner. The latter is charged with communicating with the client's primary physician. METHODS: We compared the functional status and satisfaction of WPP elderly enrollees with those of two sets of dually eligible controls drawn from the Medicaid waiver rosters. One set of controls came from persons in the same county who opted not to enroll in WPP. The second came from matched counties that did not have access to the WPP. Enrollees were interviewed in person. Family members were interviewed by telephone. RESULTS: The prevalence of activities of daily living (ADLs) and instrumental activities of daily living (IADLs) dependency was lower for the WPP sample than that for the controls. The pattern of unmet needs was generally comparable. About half of each sample had a written advance directive. Overall, there were few areas of significant difference in beneficiaries' satisfaction. The WPP families were more satisfied than either control group that services were provided when needed and were better coordinated. There were no significant differences in the prevalence of any aspect of care burden. CONCLUSIONS: The impact of WPP seems limited. There is some evidence that families perceive better coordinated care. A more complete evaluation will await the analysis of the differences in utilization patterns between WPP and the controls. PMID- 11909890 TI - Arterial wall production of cytokines in giant cell arteritis: results of a pilot study using human temporal artery cultures. AB - BACKGROUND: Giant cell arteritis (GCA) is a subacute periarteritis predominantly affecting segments of the external carotids of elderly patients. Vasculitic lesions in GCA samples might be characterized by in situ production of cytokines mRNA, indicative of macrophage and T-cell activation. However, whether the cytokine production of vessels with arteritis differs from that of vessels exposed to inflammatory conditions that originate peripheral to the vessel remains unknown. METHODS: We investigated cytokine and soluble receptor cytokine production in blood samples and cultures of human temporal arteries from 22 consecutive patients (mean age 77 +/- 6 years) further investigated for possible diagnosis of GCA: 7 patients had GCA and 15 had neither GCA nor vasculitis but had other inflammatory, infectious, or malignant diseases (controls). The production of cytokines and soluble cytokine receptors in the supernatants of cultures of 3-mm segments of temporal artery specimens, before and after lipopolysaccharide (LPS) stimulation (10 ng/ml and 10 microg/ml) and in serum, was quantified using sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: Cytokine production by temporal arteries increased significantly and in a dose dependent manner (p <.01) after LPS stimulation in all patients studied, suggesting that the system is methodologically functional. Despite a large interindividual variation, we found similar differences in cytokine production before and after stimulation by 10 ng/ml and 10 microg/ml LPS between both groups: temporal arteries of GCA patients produced more interleukin (IL)-1beta (p <.05) and IFNgamma (nonsignificant) and less tumor necrosis factor (TNF)alpha (p <.05) and IL-6 (nonsignificant) than temporal arteries of controls. The levels of TNFalpha (p <.05) and IL-6 soluble receptor (p <.05) were significantly lower in GCA patients as compared with controls in blood samples, whereas levels of cytokines in temporal artery and in blood samples were not significantly correlated at the individual level in both groups. CONCLUSIONS: The present pilot study, which requires further confirmation on a larger number of well-defined patients with GCA, suggests that a specific arterial cytokine production profile might exist in GCA (high IL-1beta +/- IFNgamma and low TNFalpha), addresses the question of the mechanisms by which IL-1beta and TNFalpha might be differentially regulated at the level of the arterial cell wall, and supports the view that cultures of the temporal artery might be an interesting tool for evaluating the role of cytokines in GCA pathogenesis. PMID- 11909893 TI - Mitochondrial optic neuropathies. PMID- 11909894 TI - A new antiepileptic drug. PMID- 11909895 TI - Improved antisaccade performance in schizophrenia with risperidone. PMID- 11909896 TI - Intensity of rehabilitation: some answers and more questions? PMID- 11909897 TI - Respective potencies of Botox and Dysport: a double blind, randomised, crossover study in cervical dystonia. PMID- 11909898 TI - Headache and hypertension: refuting the myth. PMID- 11909901 TI - Improved antisaccade performance with risperidone in schizophrenia. AB - OBJECTIVE: Antisaccade errors are consistently increased in schizophrenia. As they have been demonstrated only in cross sectional studies, it is unclear how they vary longitudinally or with different medications. In a previous cross sectional study, we reported a trend towards a reduction in error rates in a patient group treated with risperidone, compared with clozapine and sulpiride treated groups. METHODS: Gap random and antisaccade paradigms were performed on two occasions in the same sample of DSM-IV schizophrenic patients (n=12) in transition between conventional antipsychotic drugs and risperidone. A cross over design was used with six patients switching from risperidone to conventional (group I) and six in the opposite direction (group II). A control sample (n=12) was also tested on two occasions and their performance compared. The effects of practice between first and second testing and of switching between conventional antipsychotic drugs and risperidone and vice versa was also evaluated. RESULTS: A significant reduction in error rate was demonstrated during risperidone treatment (n=12), compared with conventional APD treatment. Switching from conventional to risperidone produced a reduction in errors, and vice versa. CONCLUSIONS: Treatment with risperidone was associated with improvement in antisaccade errors. PMID- 11909899 TI - Functional brain mapping of psychopathology. AB - In this paper, we consider the impact that the novel functional neuroimaging techniques may have upon psychiatric illness. Functional neuroimaging has rapidly developed as a powerful tool in cognitive neuroscience and, in recent years, has seen widespread application in psychiatry. Although such studies have produced evidence for abnormal patterns of brain response in association with some pathological conditions, the core pathophysiologies remain unresolved. Although imaging techniques provide an unprecedented opportunity for investigation of physiological function of the living human brain, there are fundamental questions and assumptions which remain to be addressed. In this review we examine these conceptual issues under three broad sections: (1) characterising the clinical population of interest, (2) defining appropriate levels of description of normal brain function, and (3) relating these models to pathophysiological conditions. Parallel advances in each of these questions will be required before imaging techniques can impact on clinical decisions in psychiatry. PMID- 11909902 TI - Structural MRI of the brain in presumed carriers of genes for schizophrenia, their affected and unaffected siblings. AB - BACKGROUND: Schizophrenia is a highly heritable disorder associated with structural brain abnormalities. The aim of this study was to establish if the gene(s) for schizophrenia are associated with specific abnormalities of brain structure. SUBJECTS: Six sibships from multiple affected families were recruited. Each sibship consisted of one patient with schizophrenia, one "obligate carrier" without the disorder but with an affected child, and one "non-affected non carrier". Such sibships are very rare, but present a powerful opportunity to separate the associations of genotype and phenotype. Obligates presumably have the gene(s) but not the disorder, affected siblings have both, whereas non affected non-carrier siblings have neither. METHOD: Brain MRI was conducted with a semiautomated region of interest analysis. The risk of false positive findings was reduced by collapsing brain regions and sides into five regions and comparing groups by repeated measures analysis of variance. RESULTS: In terms of whole brain volumes and volumes of cortical structures, obligates resembled their non affected non-carrier siblings, both groups having significantly greater volumes than their schizophrenic siblings (p=0.01 and p=0.04). Obligates also had significantly smaller ventricles than their schizophrenic siblings (p=0.03). However, with respect to the amygdalohippocampal complex, the obligates' brains resembled those of their schizophrenic siblings, both groups showing a significant reduction in volume when compared with their non-affected non-carrier siblings (p=0.001). CONCLUSIONS: In the families studied, reductions in volumes of cortical structures and reductions in whole brain volume seem to be associated with the phenotype of schizophrenia. By contrast, reduced volume of the amygdalohippocampal complex seems to be associated with genetic risk for the disorder even in the absence of disease. PMID- 11909900 TI - Intravenous immunoglobulin in neurological disease: a specialist review. AB - Treatment of neurological disorders with intravenous immunoglobulin (IVIg) is an increasing feature of our practice for an expanding range of indications. For some there is evidence of benefit from randomised controlled trials, whereas for others evidence is anecdotal. The relative rarity of some of the disorders means that good randomised control trials will be difficult to deliver. Meanwhile, the treatment is costly and pressure to "do something" in often distressing disorders considerable. This review follows a 1 day meeting of the authors in November 2000 and examines current evidence for the use of IVIg in neurological conditions and comments on mechanisms of action, delivery, safety and tolerability, and health economic issues. Evidence of efficacy has been classified into levels for healthcare interventions (tables 1 and 2). PMID- 11909903 TI - Respective potencies of Botox and Dysport: a double blind, randomised, crossover study in cervical dystonia. AB - OBJECTIVES: Botulinum toxin type A is a potent neuromuscular paralyzing agent used in various disorders including cervical dystonia. Two preparations of botulinum toxin are now commercially available ( Dysport and Botox), but much controversy remains about their respective potencies. The aim of the study was to compare the efficacy of Botox with two different ratios of Dysport. METHODS: A double blind, randomised, three period cross over study involving 54 patients with cervical dystonia was performed. The patients received the following treatments in a randomised order: Botox at the usually effective dose, Dysport at a dose of 1:3 (conversion factor of 3 between Botox and Dysport units-that is, one Botox unit=three Dysport units) and at a dose of 1:4 (conversion factor of four). The improvement of the Tsui (primary outcome criteria) and of the TWSTRS pain scales between baseline and a control visit 1 month after each of the three injections, as well as the incidence of adverse events, were assessed. RESULTS: Comparison of the Tsui scores and of the TWSTRS pain scores showed a better effect on impairment and pain with Dysport 1:3 (p=0.02 and 0.04, respectively) and 1:4 (p=0.01 and 0.02, respectively) than with Botox. The number of adverse events was higher with both Dysport treatments. The most frequent adverse event was dysphagia, found in 3%, 15.6%, and 17.3% (Botox, Dysport 1:3 and 1:4, respectively) of the patients. No adverse event required withdrawal of therapy or specific management. CONCLUSIONS: Dysport 1:3 (and Dysport 1:4 to a greater extent) is more efficient than Botox for both impairment and pain in cervical dystonia although with a somewhat higher incidence of minor adverse effects. This strongly suggests that the most appropriate conversion factor between Botox and Dysport units is less than 3 in cervical dystonia. PMID- 11909904 TI - Blood pressure and risk of headache: a prospective study of 22 685 adults in Norway. AB - OBJECTIVES: Prevalence studies of the association between blood pressure and headache have shown conflicting results. The aim was to analyse the relation between blood pressure and risk of headache in a prospective study. METHODS: A total of 22 685 adults not likely to have headache, had their baseline blood pressure measured in 1984-6, and responded to a headache questionnaire at follow up 11 years later (1995-7). The relative risk of headache (migraine or non migrainous headache) was estimated in relation to blood pressure at baseline. RESULTS: Those with a systolic blood pressure of 150 mm Hg or higher had 30% lower risk (risk ratio (RR)=0.7, 95% CI 0.6-0.8) of having non-migrainous headache at follow up compared with those with systolic pressure lower than 140 mm Hg. For diastolic blood pressure, the risk of non-migrainous headache decreased with increasing values, and these findings were similar for both sexes, and were not influenced by use of antihypertensive medication. For migraine, there was no clear association with blood pressure. CONCLUSION: In the first prospective study of blood pressure and the risk of headache, high systolic and diastolic pressures were associated with reduced risk of non-migrainous headache. One possible explanation may be the phenomenon of hypertension associated hypalgesia, which probably involves the baroreflex system influencing nociception in the brain stem or spinal cord. PMID- 11909905 TI - Dynamic cerebral autoregulation and beat to beat blood pressure control are impaired in acute ischaemic stroke. AB - OBJECTIVES: Hypertension and chronic cerebrovascular disease are known to alter static cerebral autoregulation (CA) but the effects of acute stroke on dynamic CA (dCA) have not been studied in detail. Those studies to date measuring dCA have used sympathetically induced blood pressure (BP) changes, which may themselves directly affect dCA. This study assessed whether dCA is compromised after acute stroke using spontaneous blood pressure (BP) changes as the stimulus for the dCA response. METHODS: 56 patients with ischaemic stroke (aged 70 (SD 9) years), studied within 72 hours of ictus were compared with 56 age, sex, and BP matched normal controls. Cerebral blood flow velocity was measured using transcranial Doppler ultrasound (TCD) with non-invasive beat to beat arterial BP levels, surface ECG, and transcutaneous CO(2) levels and a dynamic autoregulatory index (dARI) calculated. RESULTS: Beat to beat BP, but not pulse interval variability was significantly increased and cardiac baroreceptor sensitivity (BRS) decreased in the patients with stroke. Dynamic CA was significantly reduced in patients with stroke compared with controls (strokes: ARI 3.8 (SD 2.2) and 3.2 (SD 2.0) for pressor and depressor stimuli respectively v controls: ARI 4.7 (SD 2.2) and 4.5 (SD 2.0) respectively (p<0.05 in all cases)). There was no difference between stroke and non-stroke hemispheres in ARI, which was also independent of severity of stroke, BP, BP variability, BRS, sex, and age. CONCLUSION: Dynamic cerebral autoregulation, as assessed using spontaneous transient pressor and depressor BP stimuli, is globally impaired after acute ischaemic stroke and may prove to be an important factor in predicting outcome. PMID- 11909906 TI - Long term effects of intensity of upper and lower limb training after stroke: a randomised trial. AB - OBJECTIVE: To assess long term effects at 1 year after stroke in patients who participated in an upper and lower limb intensity training programme in the acute and subacute rehabilitation phases. DESIGN: A three group randomised controlled trial with repeated measures was used. METHOD: One hundred and one patients with a primary middle cerebral artery stroke were randomly allocated to one of three groups for a 20 week rehabilitation programme with an emphasis on (1) upper limb function, (2) lower limb function or (3) immobilisation with an inflatable pressure splint (control group). Follow up assessments within and between groups were compared at 6, 9, and 12 months after stroke. RESULTS: No statistically significant effects were found for treatment assignment from 6 months onwards. At a group level, the significant differences in efficacy demonstrated at 20 weeks after stroke in favour of the lower limb remained. However, no significant differences in functional recovery between groups were found for Barthel index (BI), functional ambulation categories (FAC),action research arm test (ARAT), comfortable and maximal walking speed, Nottingham health profile part 1(NHP-part 1), sickness impact profile-68 (SIP-68), and Frenchay activities index (FAI) from 6 months onwards. At an individual subject level a substantial number of patients showed improvement or deterioration in upper limb function (n=8 and 5, respectively) and lower limb function (n=19 and 9, respectively). Activities of daily living (ADL) scores showed that five patients deteriorated and four improved beyond the error threshold from 6 months onwards. In particular, patients with some but incomplete functional recovery at 6 months are likely to continue to improve or regress from 6 months onwards. CONCLUSIONS: On average patients maintained their functional gains for up to 1 year after stroke after receiving a 20 week upper or lower limb function training programme. However, a significant number of patients with incomplete recovery showed improvements or deterioration in dexterity, walking ability, and ADL beyond the error threshold. PMID- 11909907 TI - Timing of surgery for supratentorial aneurysmal subarachnoid haemorrhage: report of a prospective study. AB - OBJECTIVES: The debate on the timing of aneurysm surgery after subarachnoid haemorrhage (SAH) pivots on the balance of the temporal risk for fatal rebleeding versus the risk of surgical morbidity when operating early on an acutely injured brain. By following a strict management protocol for SAH, the hypothesis has been tested that in the modern arena of treatment for aneurysmal SAH the timing of surgery to secure supratentorial aneurysms does not affect surgical outcome. METHODS: Over a 6 year period, patients admitted with a diagnosis of SAH to a regional neurosurgical unit have been prospectively studied. All have been on a management protocol in which early transfer and resuscitation has been followed regardless of age and clinical condition. Angiographic investigation and surgery have been pursued in those who have been able to at least flex to pain. A total of 1168 patients (60.7% female, mean age 54.3) with proved SAH were received on median day 1 (86.4% arrived within 3 days) of the ictus. Of these, 784 (67.1%) showed aneurysms on angiography and were prepared for surgery. Those who received surgery for a supratentorial aneurysm within 21 days of the ictus were included in the final analysis (n=550). Patients with an initial negative angiogram, with posterior circulation aneurysms, or aneurysms treated by endovascular means, with aneurysms requiring emergency surgery for space occupying haematomas, with aneurysms which re-bled before surgery, and those who received very late surgery (after 21 days from ictus) were excluded. Surgical outcomes at hospital discharge and after 6 months were assessed using the Glasgow outcome score (GOS). Discharge destination and duration of stay in a neurosurgical ward were also documented. The influence of the timing of surgery (early group day 1-3 postictus, intermediate group day 4-10, or late group day 11-21) was analysed prospectively. RESULTS: 60.2% of cases fell into the early surgery group, 32.4% into the intermediate group, and 7.5% into the late operated group. Late surgery was due to delays in diagnosis, transfer, and logistic factors, but not clinical decision. The demographic characteristics, site of aneurysm, and clinical condition of the patients at the time of initial medical assessment were balanced in the three surgical timing groups. There was no significant difference in GOS between the surgical timing groups at 6 months (favourable GOS score 4 and 5: 83.2%, 80.5%, and 83.8% respectively; p=0.47, Kruskal-Wallis test). Outcome was favourable in 84% of patients under 65 years, and 70% in those over 65. The discharge destinations (home, referring hospital, nursing home, rehabilitation centre) showed no significant difference between surgical timing groups. There was no significant difference in mean time to discharge after admission to this hospital from the referring hospital (16.2, 16.2, and 14.6 days for early, intermediate, and late groups respectively; p=0.789, Analysis of variance (ANOVA)). As a result, there was reduction in the mean duration of total hospital inpatient stay in favour of the earliest operated patients (mean time 18.1, 22.0, and 28.3 days respectively; p=0.001. ANOVA showed that besides age, the only determinant of surgical outcome and duration of stay was presenting clinical grade (p<0.0005). CONCLUSION: The current management of patients presenting with SAH from anterior circulation aneurysms allows early surgery to be followed safely regardless of age. The only independent variables affecting outcome are age and clinical grade at presentation. The timing of surgery did not significantly affect surgical outcome, promoting a policy for early surgery that avoids the known risks of rebleeding and reduces inpatient stay. PMID- 11909908 TI - Endovascular management of unruptured intracranial aneurysms: does outcome justify treatment? AB - OBJECTIVE: The appropriate management of unruptured intracranial aneurysms depends on a complete understanding of their natural history and on the risks and efficacy of treatment options. There is little current data on the risks of endovascular therapy for these aneurysms. The aim of this study was to assess outcome of endovascular treatment of unruptured intracranial aneurysms. METHOD: A retrospective analysis was performed on all unruptured aneurysms treated by Guglielmi detachable (GD) coils at this institution from 1994 to 2000. RESULTS: Seventy three unruptured aneurysms were treated in 62 patients. There were 52 female and 10 male patients, with a median age of 55.7 years. Clinical background was: subarachnoid haemorrhage due to rupture of an additional aneurysm (40), headache (4), third nerve palsy (four), familial (four), and incidental (10). There were 14 technical failures with no clinical sequelae. Four procedural complications occurred (5.5%, 95% confidence interval (95% CI) 0.3% to 10.9%). One patient had temporary clinical sequelae (1.4%, 95% CI 0% to 2.7%); 79% of treated aneurysms had stable occlusions at follow up, 10.5% showed improved occlusion grade, 10.5% showed some recurrence, and three patients have required retreatment. Follow up modified Glasgow outcome scores were grade 1, 71%; grade 2, 18%; grade 3, 3%; grade 4, 3%. There were no deaths or haemorrhages during the follow up period. Two patients died as a result of complications from subarachnoid haemorrhage. CONCLUSION: The endovascular treatment of patients with unruptured aneurysms is safe with few clinical or procedural complications. Poor outcomes were only seen in those patients who presented with subarachnoid haemorrhage due to rupture of an aneurysm at another site. PMID- 11909910 TI - Expression of chemokines in the CSF and correlation with clinical disease activity in patients with multiple sclerosis. AB - OBJECTIVE: To define the chemokine profile in the CSF of patients with multiple sclerosis (MS) and compare it with three control groups; patients with benign headache (headache), non-inflammatory neurological diseases (NIND), and other inflammatory neurological diseases (IND). In addition, the correlations of CSF chemokine concentrations with chemokine receptor expression on CSF CD4(+) T cells and with clinical disease activity were assessed. METHODS: Forty three patients with MS, 24 with IND, 44 with NIND, and 12 with benign headache undergoing diagnostic or therapeutic lumbar puncture were included. Supernatant fluid from CSF was analysed for four beta (CCL2, CCL3, CCL4, CCL5) and two alpha (CXCL9, CXCL10)chemokines by enzyme linked immunosorbent assay (ELISA). Chemokine receptors CCR3, CCR5, and CXCR3 on CD4(+) T cells from eight patients with MS were analysed using directly conjugated fluorescent labelled monoclonal antibodies and flow cytometry. RESULTS: CXCL10, formerly interferon-gamma inducible protein-10 (IP-10), was significantly increased and CCL2, formerly monocyte chemoattractant protein-1 (MCP-1), was significantly reduced in the CSF of patients with MS and IND compared with those with benign headache and NIND. Concentrations of CXCL10 were significantly greater in patients with relapsing remitting compared with secondary progressive MS and correlated significantly with CXCR3 expression on CSF CD4(+) T cells from patients with MS. Concentrations of CXCL10 decreased and CCL2 concentrations increased as time from the last relapse increased in patients with MS. CONCLUSION: Increased CXCL10 and decreased CCL2 concentrations in the CSF are associated with relapses in MS. Although serial values from individual patients were not available, this study suggests that CXCL10 and CCL2 may return towards baseline concentrations after a relapse. Correlation of CXCL10 with CD4(+) T cell expression of CXCR3 was consistent with its chemoattractant role for activated lymphocytes. Thus CXCL10 neutralising agents and CXCR3 receptor antagonists may be therapeutic targets in MS. PMID- 11909909 TI - Medial temporal lobe atrophy predicts Alzheimer's disease in patients with minor cognitive impairment. AB - OBJECTIVES: To investigate whether medial temporal lobe atrophy predicted outcome in patients with minor cognitive impairment and whether assessment of the medial temporal lobe could increase the predictive accuracy of age and delayed recall for outcome. Quantitative and qualitative methods of assessing the medial temporal lobe were also compared. METHODS: Patients with minor cognitive impairment older than 50 years (n=31) were selected from a memory clinic and were followed up for on average 1.9 years. The medial temporal lobe was assessed in three different ways: volumetry of the hippocampus, volumetry of the parahippocampal gyrus, and qualitative rating of medial temporal lobe atrophy (MTA). Outcome measures were Alzheimer type dementia or cognitive decline at follow up. Delayed recall was tested with a verbal learning test. RESULTS: Ten patients had experienced cognitive decline at follow up, of whom seven had probable Alzheimer type dementia. All medial temporal lobe measurements were associated with cognitive decline at follow up (p trend analysis between 0.001 (hippocampus) and 0.05 (parahippocampal gyrus)). Only the hippocampal volume and MTA score were associated with Alzheimer type dementia at follow up (p trend analysis respectively 0.003 and 0.01). All medial temporal lobe measurements increased the predictive accuracy of age and the delayed recall score for cognitive decline (p increase in predictive accuracy varied between <0.001 (hippocampus) and 0.02 (parahippocampal gyrus and MTA score)) and the hippocampal volume and the MTA score increased the predictive accuracy of age and the delayed recall score for Alzheimer type dementia (p= 0.02). CONCLUSIONS: The ability to detect patients at high risk for Alzheimer type dementia among those with minor cognitive impairment increases when data on age and memory function are combined with measures of medial temporal lobe atrophy. Volumetry of the hippocampus is preferred, but qualitative rating of medial temporal lobe atrophy is a good alternative. PMID- 11909912 TI - Long term course of childhood epilepsy following relapse after antiepileptic drug withdrawal. AB - OBJECTIVE: To explore the course of epilepsy following relapse after antiepileptic drug (AED) withdrawal. METHODS: Forty two patients were identified with onset of epilepsy in childhood in whom AEDs had been withdrawn after at least 2 years of seizure freedom, and in whom a relapse had occurred. Two patients were lost to follow up. RESULTS: Median follow up after AED withdrawal was 5.9 years (range 1.6-13.2 years). Relapse occurred in more than half of the patients within 6 months of AED withdrawal. At the end of follow up, 12 patients (30%) were seizure free for at least 1 year (mean 10.4 years) without medication; 16 (40%) were seizure free for at least 1 year (mean 5.3 years) with ongoing medication; and 12 patients (30%) were seizure free for less than 1 year with medication. No status epilepticus occurred in any patient after withdrawal. Age at onset, if over the age of 5, combined with normal intelligence were predictive of an excellent outcome; presence of a neurological disorder, and hence symptomatic aetiology, was predictive of poor outcome after a relapse. CONCLUSIONS: Fears that premature withdrawal of AEDs might result in uncontrollable seizures were unsubstantiated in this study. The current practice of withdrawing AEDs in children who have been seizure free for 2 years can be beneficial to most of these patients. PMID- 11909911 TI - The value of temporary external lumbar CSF drainage in predicting the outcome of shunting on normal pressure hydrocephalus. AB - OBJECTIVE: It has been reported that temporary external lumbar CSF drainage (ELD) is a very accurate test for predicting the outcome after ventricular shunting in patients with normal pressure hydrocephalus (NPH). However, only a limited number of patients have been studied for assessing the predictive accuracy of ELD. Therefore, the value of ELD in predicting the outcome after a ventriculoperitoneal shunt in patients with presumed NPH was assessed. METHODS: All patients with presumed NPH were invited to participate in this study. Clinical assessment, MRI, and neuropsychological evaluation were followed by a lumbar CSF tap test consisting of removing 40 ml CSF. When this test resulted in marked clinical improvement of gait impairment, mental disturbances, or both, the patient was shunted without further tests. In patients with either questionable or no improvement after the CSF tap test, ELD was carried out. The value of ELD for predicting the outcome after shunting was calculated by correlating the results of ELD with that of ventriculoperitoneal shunting. RESULTS: Between January 1994 and December 2000, 49 presumed NPH patients from three institutes were included. Forty three had idiopathic, and the remaining six had secondary NPH. Forty eight patients were shunted; 39 had an ELD of whom 38 completed the test. After 2 months 35 of the 48 (73%) shunted patients had improved. The predictive value of a positive ELD was 87% (95% confidence interval (95% CI) 62 98) and that of a negative ELD 36% (95% CI 17-59). In two patients serious test related complications (meningitis) occurred without residual deficit. CONCLUSION: The study suggests that although the predictive value of a positive ELD is high, that of a negative ELD is deceptively low because of the high rate of false negative results. The costs and invasiveness of the test and the possibility of serious test related complications further limits its usefulness in managing patients with presumed NPH. PMID- 11909913 TI - Long term reshaping of language, sensory, and motor maps after glioma resection: a new parameter to integrate in the surgical strategy. AB - OBJECTIVES: To describe cortical reorganisation and the effects of glioma infiltration on local brain function in three patients who underwent two operations 12-24 months apart. METHODS: Three patients who had no neurological deficit underwent two operations for low grade glioma, located in functionally important brain regions. During each operation, local brain function was characterised by electrical mapping and awake craniotomy. RESULTS: Language or sensorimotor areas had been invaded by the tumour at the time of the first operation, leading to incomplete glioma removal in all cases. Because of a tumour recurrence, the patients were reoperated on between 12 and 24 months later. Functional reorganisation of the language, sensory, and motor maps was detected by electrical stimulation of the brain, and this allowed total glioma removal without neurological sequelae. CONCLUSIONS: These findings show that surgical resection of a glioma can lead to functional reorganisation in the peritumorous and infiltrated brain. It may be that this reorganisation is directly or indirectly caused by the surgical procedure. If this hypothesis is confirmed by other studies, the use of such brain plasticity potential could be used when planning surgical options in some patients with low grade glioma. Such a strategy could extend the limits of tumour resection in gliomas involving eloquent brain areas without causing permanent morbidity. PMID- 11909915 TI - Abnormal visual projection in a human albino studied with functional magnetic resonance imaging and visual evoked potentials. AB - The albino visual pathway is abnormal in that many fibres from the temporal retina project to the contralateral visual cortex. The visual projections in a human albino and a control have been investigated with fMRI and VEP during independent visual stimulation of both hemifields. Activity in the occipital cortex in the normal was contralateral to the stimulated visual field, whereas it was contralateral to the stimulated eye in the albino, independent of the stimulated visual field. Thus, the albino visual cortex is activated not only by stimulation in the contralateral visual field, but also by abnormal input representing the ipsilateral visual field. These novel findings help elucidate the nature of albino misrouting. PMID- 11909916 TI - Nikola Tesla (1856-1943). PMID- 11909914 TI - Concurrent excitatory and inhibitory effects of high frequency stimulation: an oculomotor study. AB - OBJECTIVE: To describe a reversible neurological condition resembling a crossed midbrain syndrome resulting from high frequency stimulation (HFS) in the midbrain. METHODS: Postoperative evaluation of quadripolar electrodes implanted in the area of the subthalamic nucleus of 25 patients with Parkinson's disease (PD) successfully treated by HFS. RESULTS: Four of the 25 patients experienced reversible acute diplopia, with dystonic posture and tremor in the contralateral upper limb when the white matter between the red nucleus and the substantia nigra was stimulated. The motor signs resembled those caused by lesions of the red nucleus. The ipsilateral resting eye position was "in and down" (three patients) or "in" (one patient). Enophthalmos was seen. Abduction was impaired and vertical eye movements were limited, but adduction was spared. The movements of the controlateral eye were normal. The ocular signs could be best explained by sustained hyperactivity of the extrinsic oculomotor nerve. Simultaneous tonic contraction of the superior rectus, the inferior rectus, and inferior oblique may cause the enophthalmos and partial limitation of upward and downward eye movements. Antagonist tonic contraction of the ipsilateral medial rectus severely impairs abduction. CONCLUSION: This crossed midbrain syndrome, possibly resulting from simultaneous activation of oculomotor nerve and lesion-like inhibition of the red nucleus suggests that high frequency stimulation has opposite effects on grey and white matter. PMID- 11909917 TI - Ataxia in the setting of complicated enteropathy: double jeopardy. AB - The differential diagnosis of subacute onset ataxia in the setting of enteropathy is wide. A 54 year old patient with a pancerebellar syndrome and known ulcerative jejunoileitis is described. Small bowel biopsy showed evidence of enteropathy associated T cell lymphoma and subsequent neuropathological analysis and immunophenotyping confirmed metastasis of this tumour to the cerebellum. The presence of anti-gliadin antibodies and MRI evidence of a more longstanding process suggested additional immunologically mediated cerebellar dysfunction. Lymphomatous involvement of the CNS is rare in patients with complicated enteropathies, and has not been previously reported to involve the cerebellar parenchyma. This diagnostic possibility should be borne in mind before attributing cerebellar dysfunction in patients with the coeliac related enteropathies to nutritional compromise or immunological dysfunction (gluten ataxia) alone. PMID- 11909919 TI - Isolated corpus callosal infarction secondary to pericallosal artery disease presenting as alien hand syndrome. AB - Two patients are described with the callosal type of alien hand syndrome. Both presented with abnormal feelings in the left upper limb and intermanual conflict without clinical evidence of callosal apraxia or frontal lobe dysfunction such as motor deficit or reflexive grasping. Imaging studies disclosed subacute infarction in the body and splenium of the corpus callosum due to pericallosal artery disease. These patients were unique in their presentation as a callosal type of alien hand syndrome secondary to ischaemic stroke. PMID- 11909918 TI - Separating depersonalisation and derealisation: the relevance of the "lesion method". AB - OBJECTIVES: Depersonalisation (DP) and derealisation (DR) are often met with in patients with a wide range of localisable neurological conditions. This suggests that the "lesion method" might be a valid approach to study the neurobiology of DP/DR. However, the fact that anxiety can trigger DP/DR makes it difficult to establish whether the presence of DP/DR in neurological patients is mainly determined by coexisting anxiety or by lesion location. To overcome this difficulty, we suggest the study of neurological phenomena, which although not considered as DP/DR, bear enough phenomenological resemblance with them as to warrant their use as models. METHODS: One patient with "visual hypoemotionality" and another with "hemiasomatognosia" are described in detail together with a selective literature review. RESULTS: Complaints of patients with visual hypoemotionality are indistinguishable from those of patients with "visual derealisation". There is also a phenomenological overlap between "asomatognosia" and the symptom of "body alienation", which is a central feature of depersonalisation. CONCLUSIONS: Phenomenological similarities between visual hypoemotionality and DR suggest that a disruption of the process by means of which perception becomes emotionally coloured may be an underlying mechanism in both conditions. Likewise, phenomenological overlaps with asomatognosia suggest that DP might result from parietal mechanisms disrupting the experience of body ownership and agency. These findings give validity to the notion that DP and DR may have distinct neurobiological mechanisms. PMID- 11909920 TI - A Raymond Chandler trio. PMID- 11909922 TI - Acute small fibre sensory neuropathy: another variant of Guillain-Barre syndrome? AB - Six patients who presented with acute sensory neuropathy were studied. All patients underwent detailed clinical assessment along with electrophysiological tests and relevant laboratory investigations. All patients had acute onset numbness, reaching the peak deficit within 4 weeks. Four of them had associated burning dysaesthesia. An antecedent illness was reported in four; diarrhoea in three, and urinary tract infection in one. The neurological examination disclosed normal muscle strength, symmetric glove and stocking type sensory loss for pain and temperature, normal proprioception, and vibration senses with normal or brisk tendon reflexes. Analysis of CSF demonstrated albuminocytological dissociation in all. Routine motor and sensory nerve conduction studies were normal. Sympathetic skin responses were also normal except for the lower limbs in one patient. Stool cultures for Campylobacter jejuni were negative. The outcome was favourable. Burning dysaesthesia disappeared within 4 months. Numbness and objective sensory loss tended to persist longer. The clinical features and normal routine nerve conduction studies, which assess large diameter nerve fibre function, indicate small sensory fibre dysfunction in the group. Their presentation and CSF findings would fit into the diagnosis of sensory Guillain-Barre syndrome. The current study suggests that acute small fibre sensory neuropathy (ASFSN) is another clinical entity which could perhaps be included in the heterogeneous range of Guillain-Barre syndrome. PMID- 11909921 TI - Intrinsic spinal cord lesions complicating epidural anaesthesia and analgesia: report of three cases. AB - Serious neurological complications related to epidural anaesthesia and analgesia are only rarely reported. We describe the clinical and radiological features of three patients who sustained intrinsic spinal cord lesions after attempted epidural catheterisation. In each case there was an early onset of motor and sensory impairment after the procedure and MRI demonstrated similar, extensive, paracentral, high signal intensity lesions within the cord on T2 weighted images. Possible mechanisms to explain these MRI appearances are discussed. It is proposed that the most likely cause of these lesions was direct trauma to the spinal cord during the procedure and subsequent injection of fluid into the spinal cord producing localised hydromyelia. The prognosis in each case was for a gradual recovery of motor function but spinothalamic sensory impairment and severe spontaneous pain over the affected area persisted. PMID- 11909923 TI - A case of aceruloplasminaemia: abnormal serum ceruloplasmin protein without ferroxidase activity. AB - A 34 year old diabetic man with a complete deficiency of serum ferroxidase activity, regardless of the presence of serum ceruloplasmin (Cp), a multicopper ferroxidase protein, is described. The patient had had diabetes mellitus for 13 years, and was also found to have retinal degeneration accompanied by the development of a hearing disturbance of unknown aetiology. Laboratory examination showed markedly increased serum ferritin and low serum iron. Magnetic resonance imaging showed a pronounced hypointensity in the putamen, caudate, cerebellar dentate, and thalamus on T2 weighted images, and also disclosed a low level signal in the liver, suggesting the accumulation of some magnetic substances in the brain and liver. Liver biopsies histochemically identified iron deposition in the hepatocytes. Most of these findings were consistent with the newly established autosomal recessive disease "aceruloplasminaemia", except for the presence of serum Cp and the lack of apparent neurological symptoms. Interestingly, no ferroxidase activity was detected in the patient's serum, whereas suppressed ferroxidase activity was found in his mother's serum. A nucleotide sequence analysis of the Cp gene showed two mutations; a C to T substitution at nucleotide 2701 in exon 16, resulting in a nonsense mutation at amino acid 882 (Arg882ter), and a T to G substitution at nucleotide 2991 in exon 17, resulting in an amino acid alternation at amino acid 978 (His978Gln). The second mutation was also found in the patient's mother. The absence of serum ferroxidase activity despite the presence of serum Cp protein in this compound heterozygote was considered to be due to the production of a non-functional Cp harbouring no ferroxidase activity. PMID- 11909924 TI - Deep brain stimulation of the centre median-parafascicular complex in patients with movement disorders. AB - The centre median-parafascicular (CM-Pf) complex of the thalamus is considered to be a possible target for deep brain stimulation (DBS) in patients with movement disorders. In a prospective study on the effect of CM-Pf DBS versus somatosensory thalamic DBS on chronic neuropathic pain, three of 12 patients had additional movement disorders. Bifocal quadripolar electrodes were implanted by computed tomography guided stereotactic surgery under local anaesthesia contralaterally to the side of the pain for test stimulation. Two of the three patients with movement disorders had permanent implantation of CM-Pf electrodes. During test stimulation of the left CM-Pf complex for several days, a 67 year old woman received no benefit with respect to the neuropathic pain, but the choreoathetotic movements of her right foot ceased. As the pain syndrome was not improved, she decided not to have permanent implantation. A 74 year old man with postzoster neuralgia and allodynia enjoyed excellent relief from his pain with chronic CM-Pf DBS. In addition, improvement in the tremor at rest was noted. A 72 year old man had sustained reduction in his stump dyskinesias. Further evaluation of the possible role of the "forgotten" central and medial thalamic nuclei in the treatment of movement disorders may be warranted. PMID- 11909925 TI - Outpatient video EEG recording in the diagnosis of non-epileptic seizures: a randomised controlled trial of simple suggestion techniques. AB - OBJECTIVE: To assess the yield of recorded habitual non-epileptic seizures during outpatient video EEG, using simple suggestion techniques based on hyperventilation and photic stimulation. DESIGN: Randomised controlled trial of "suggestion" v "no suggestion" during outpatient video EEG recording. SETTING: Regional epilepsy service (tertiary care; single centre). PARTICIPANTS: 30 patients (22 female, 8 male), aged over 16 years, with a probable clinical diagnosis of non-epileptic seizures; 15 were randomised to each group. MAIN OUTCOME MEASURES: Yield of habitual non-epileptic seizures recorded, and requirement for additional inpatient video EEG. RESULTS: 10/15 patients had habitual non-epileptic seizures with suggestion; 5/15 had non-epileptic seizures with no suggestion (p = 0.058; NS); 8/9 patients with a history of previous events in medical settings had non-epileptic seizures recorded during study. Logistic regression analysis with an interaction clause showed a significant effect of suggestion in patients with a history of previous events in medical settings (p = 0.003). An additional inpatient video-EEG was avoided in 14 of the 30 patients (47%). CONCLUSIONS: Habitual non-epileptic seizures can be recorded reliably during short outpatient video EEG in selected patients. Simple (non invasive) suggestion techniques increase the yield at least in the subgroup with a history of previous events in medical settings. Inpatient video EEG can be avoided in some patients. PMID- 11909926 TI - Vertebral artery dissection presenting as neuralgic amyotrophy. PMID- 11909927 TI - Postoperative pseudoaneurysm of the superficial temporal artery. PMID- 11909928 TI - Restless legs syndrome and peripheral neuropathy. PMID- 11909929 TI - Parent reports of coping and stress responses in children with recurrent abdominal pain. AB - OBJECTIVE: To examine relationships among coping, stress responses, pain, somatic symptoms, and anxious/depressed symptoms in a sample of children and adolescents with recurrent abdominal pain (RAP). METHOD: We assessed parents' reports of coping and involuntary responses to stress in relation to pain, somatic symptoms, and symptoms of anxiety and depression in a sample of 174 children and adolescents with RAP. RESULTS: Based on parent reports, children's primary control engagement coping (e.g., problem solving, emotional modulation) and secondary control engagement coping (e.g., acceptance, distraction, positive thinking) in response to pain were associated with fewer somatic complaints and symptoms of anxiety and depression; secondary control engagement coping was also associated with less pain. Involuntary engagement (e.g., physiological reactivity, rumination) and disengagement (e.g., escape, inaction) responses to pain were associated with more somatic symptoms and higher levels of anxiety and depression. CONCLUSIONS: We highlight implications of these findings for understanding processes of coping and stress reactivity in children with RAP. PMID- 11909930 TI - Predicting children's response to an invasive medical investigation: the Influence of effortful control and parent behavior. AB - OBJECTIVE: To investigate the relative contributions of effortful control (reflecting the child's ability to shift and refocus attention) and parental coping- and distress-promoting behaviors to children's coping and distress during the voiding cystourethrogram (VCUG, X-ray of the kidneys). METHOD: Thirty-two children between ages 2 and 7 years were videotaped undergoing the VCUG. Parent and child behaviors were coded according to the CAMPIS-R (Blount et al., 1997), and parents completed a temperament inventory assessing effortful control across a range of everyday situations. RESULTS: Children manifested relatively high rates of distress and low rates of coping. Their coping attempts were not associated with reduced rates of distress. The most frequent child coping behavior was distraction. Both effortful control and parent coping-promoting behavior (particularly talk about topics other than the VCUG) made independent contributions to child coping behavior. Parent distress-promoting behavior (particularly reassurance) made a strong contribution to child distress behavior. CONCLUSIONS: Factors relating to the child (effortful control) and parent (coping and distress-promoting behaviors) both contribute to children's response to an aversive medical procedure. Interventions that facilitate parent coping and promoting behavior, reduce their distress-promoting behavior, and compensate for children's infrequent and ineffective use of coping strategies (such as distraction) may be optimal for young children, particularly those low in effortful control. PMID- 11909931 TI - Neurocognitive development of young children with sickle cell disease through three years of age. AB - OBJECTIVE: To determine (1) the neurocognitive development of children with sickle cell disease (SCD) from 6 months through 36 months of age, (2) the independent and combined contributions of biomedical risk and parenting risk to child neurocognitive functioning, and (3) the independent and combined contributions of biomedical risk, parent cognitive processes, and family functioning to parent adjustment. METHOD: The study sample included 89 African American children and their parents served through the Duke University-University of North Carolina Comprehensive Sickle Cell Center. Measures of cognitive and psychomotor development were obtained at 6, 12, 24, and 36 months of age, and parents completed self-report measures of the cognitive processes of daily stress and attributional style, psychological adjustment, and family functioning. RESULTS: There was no significant decrease in psychomotor functioning (PDI) over time but cognitive functioning (MDI) declined, with a significant decrease occurring between the 12- and 24-month assessment points. At 24 months, poorer cognitive functioning was associated with parenting risk, in terms of a learned helplessness attributional style, and biomedical risk, in terms of HbSS phenotype. Levels of psychological distress within the clinical range were reported by 24% of the parents, and poorer parent adjustment was associated with high levels of daily stress, less knowledge about child development, lower expectations of efficacy, and HbSC phenotype. CONCLUSIONS: The findings indicate that young children with SCD are at risk for neurocognitive impairment and provide support for the initiation of early intervention studies to promote neurocognitive development. PMID- 11909932 TI - Interactive effects of internalizing and externalizing problem behaviors on recurrent pain in children. AB - OBJECTIVE: To examine, in children, relationships between self-reported recurrent pain and emotion regulation indicated by rated internalizing and externalizing problem behaviors and adjustment. METHOD: Finnish 11-12-year-old schoolchildren (N = 414) completed a questionnaire measuring recurrent pain. Emotion regulation was assessed by a Multidimensional Peer Nomination Inventory, Teacher Rating Form. Relationships between recurrent pain and emotion regulation were examined in logistic regression analyses, after controlling for past injuries and chronic illnesses. RESULTS: Independent of injuries and chronic illnesses, externalizing and internalizing problem behaviors related to recurrent pain, and more so together than separately. Gender differences were found; constructive behavior associated with recurrent pain only in girls. CONCLUSIONS: Low self-control of emotions, indicated by internalizing and externalizing problem behaviors, was related to pain in both boys and girls; high self-control of emotions, indicated by constructive behavior, associated with pain only in girls. PMID- 11909933 TI - Behavioral outcome of preschoolers exposed prenatally to cocaine: role of maternal behavioral health. AB - OBJECTIVE: To examine the impact of prenatal cocaine exposure and maternal behavioral health (recent drug use and psychological functioning) on child behavior at age 5 years. METHOD: In this longitudinal investigation, maternal report of child behavior was assessed using the Achenbach Child Behavior Checklist (CBCL) in 140 cocaine-exposed and 181 noncocaine-exposed (61 alcohol, tobacco, and/or marijuana-exposed, and 120 nondrug-exposed) low-income, African American children. Structural equation modeling was used to estimate suspected causal relationships between indicators of maternal behavioral health at 5-year follow-up, according to self-report on a modified Addiction Severity Index (ASI) and CBCL scores. RESULTS: Prenatal cocaine exposure was not related to child behavior at age 5. Recent maternal drug use and psychological functioning had relationships with CBCL Internalizing and Externalizing scores. However, when considered within a combined model, only maternal psychological functioning remained significant. CONCLUSIONS: Findings highlight the importance of maternal functioning in the behavioral outcome of children exposed prenatally to cocaine. PMID- 11909934 TI - Family adjustment to childhood developmental disability: a measure of parent appraisal of family impacts. AB - OBJECTIVE: To develop the Family Impact of Childhood Disability Scale (FICD) to assess subjective interpretation or "primary appraisal" of parents regarding the impact of a child with developmental disabilities on the family. METHOD: A random sample of 87 families was assessed while children with developmental disabilities were in the preschool years. After 7 years had elapsed, 64 of these families were interviewed again when the children were in the preteen years. A set of standardized self-report measures provided mother and father views of child, parent, and family functioning. RESULTS: The FICD demonstrated adequate internal consistency, with some evidence of discriminant and predictive validity. The FICD total score, based on the discrepancy between positive and negative subscale scores, was found to be a significant predictor of future parenting stress of mothers and of fathers, even when controlling for other important explanatory variables such as marital adjustment and level of disability in a child. CONCLUSIONS: The 15-item FICD offers a brief assessment of both positive and negative parent appraisals, with a total discrepancy score that predicts long term parenting stress. PMID- 11909935 TI - Hospital emergency rooms and children's health care attitudes. AB - OBJECTIVE: To assess attitudes of children requiring hospital emergency room (ER) treatment for trauma injuries 5 years afterward to evaluate the long-term effect of treatment distress. For comparison, health care attitudes of a large random sample of children were assessed. METHOD: Children (N = 139, 7-19 years old) recruited from the ER completed a health care attitude questionnaire. Comparable schoolchildren (N = 1,300) completed the same questionnaire, with the addition of a few questions asking about hospital contact. The ER-recruited group was part of a 5-year follow-up study, and at the time of initial recruitment, their parents had rated their children's degree of distress at both the time of injury and of ER treatment on a 6-point scale. RESULTS: For the ER-recruited sample, the degree of distress during ER treatment did not seem to have longterm effects on children's attitudes. For the random sample, contact with the ER, especially for a trauma injury, was related to children liking the ER more. CONCLUSIONS: Although other research has shown that aversive medical experiences may negatively affect children's attitudes, these findings suggest that the nature of the medical contact is important in how children interpret medically induced pain, which is related to their attitudes. PMID- 11909936 TI - The impact of maternal behavior on children's pain experiences: an experimental analysis. AB - OBJECTIVE: To provide an experimental investigation of the impact of maternal behavior on children's pain experiences. METHOD: Participants were 120 healthy children (60 boys, 60 girls) between the ages of 8 and 12 years and their mothers. Mothers were randomly assigned and trained to interact with their children in one of three ways while the children were exposed to lab-induced cold pressor pain: (1) a pain-promoting interaction, (2) a pain-reducing interaction, and (3) a no training control group. Training was based on behaviors presumed to have the expected impact, as based on correlational studies reported in the literature. Children's pain experiences during the cold pressor were assessed using self-reports of intensity and affect, coding of facial activity, tolerance, and heart rate responsiveness. RESULTS: Girls whose mothers interacted with them in the pain-promoting manner reported more pain than daughters of mothers in the control group, who in turn reported more pain than girls whose mothers interacted with them in the pain-reducing manner. This effect was not significant for boys. Maternal interaction type had no effect on children's pain affect, facial activity, tolerance, or heart rate. CONCLUSIONS: Results indicate that maternal behavior can have a direct impact on their daughters' subjective reports of pain. These data support the importance of social learning factors in influencing children's pain experiences. PMID- 11909937 TI - Brief report: Assessment of children's gastrointestinal symptoms for clinical trials. AB - OBJECTIVE: To conduct a pilot study evaluating a procedure for assessment of daily symptoms and functioning in pediatric patients. METHOD: Participants included 11 parent-child dyads referred to a tertiary care center for evaluation of constipation and abdominal pain. Each family was provided a hand-held computer and modem. For 7 consecutive days, parents and children (ages 6-10 years) responded as a team to questions regarding the level of children's gastrointestinal symptoms and the extent to which symptoms interfered with the day's activities. Parents responded to a telephone interview evaluating the procedure. RESULTS: Parents reported that children understood most questions and that responses entered into the computer were accurate. Parents and children were enthusiastic about the data collection method. Some technical problems arose in use of the computers. CONCLUSIONS: Within the limitations of a small sample, this data collection procedure appears to have promise for evaluating pediatric symptom outcomes. PMID- 11909938 TI - SRC catalytic but not scaffolding function is needed for integrin-regulated tyrosine phosphorylation, cell migration, and cell spreading. AB - Src family kinases (SFKs) are crucial for signaling through a variety of cell surface receptors, including integrins. There is evidence that integrin activation induces focal adhesion kinase (FAK) autophosphorylation at Y397 and that Src binds to and is activated by FAK to carry out subsequent phosphorylation events. However, it has also been suggested that Src functions as a scaffolding molecule through its SH2 and SH3 domains and that its kinase activity is not necessary. To examine the role of SFKs in integrin signaling, we have expressed various Src molecules in fibroblasts lacking other SFKs. In cells plated on fibronectin, FAK could indeed autophosphorylate at Y397 independently of Src but with lower efficiency than when Src was present. This step was promoted by kinase inactive Src, but Src kinase activity was required for full rescue. Src kinase activity was also required for phosphorylation of additional sites on FAK and for other integrin-directed functions, including cell migration and spreading on fibronectin. In contrast, Src mutations in the SH2 or SH3 domain greatly reduced binding to FAK, Cas, and paxillin but had little effect on tyrosine phosphorylation or biological assays. Furthermore, our indirect evidence indicates that Src kinase activity does not need to be regulated to promote cell migration and FAK phosphorylation. Although Src clearly plays important roles in integrin signaling, it was not concentrated in focal adhesions. These results indicate that the primary role of Src in integrin signaling is as a kinase. Indirect models for Src function are proposed. PMID- 11909939 TI - Mutation of mouse p53 Ser23 and the response to DNA damage. AB - Recent studies have suggested that phosphorylation of human p53 at Ser20 is important for stabilizing p53 in response to DNA damage through disruption of the interaction between MDM2 and p53. To examine the requirement for this DNA damage induced phosphorylation event in a more physiological setting, we introduced a missense mutation into the endogenous p53 gene of mouse embryonic stem (ES) cells that changes serine 23 (S23), the murine equivalent of human serine 20, to alanine (A). Murine embryonic fibroblasts harboring the p53(S23A) mutation accumulate p53 as well as p21 and Mdm2 proteins to normal levels after DNA damage. Furthermore, ES cells and thymocytes harboring the p53(S23A) mutation also accumulate p53 protein to wild-type levels and undergo p53-dependent apoptosis similarly to wild-type cells after DNA damage. Therefore, phosphorylation of murine p53 at Ser23 is not required for p53 responses to DNA damage induced by UV and ionizing radiation treatment. PMID- 11909940 TI - Analysis of sequence upstream of the endogenous H19 gene reveals elements both essential and dispensable for imprinting. AB - Imprinting of the linked and oppositely expressed mouse H19 and Igf2 genes requires a 2-kb differentially methylated domain (DMD) that is located 2 kb upstream of H19. This element is postulated to function as a methylation sensitive insulator. Here we test whether an additional sequence 5' of H19 is required for H19 and Igf2 imprinting. Because repetitive elements have been suggested to be important for genomic imprinting, the requirement of a G-rich repetitive element that is located immediately 3' to the DMD was first tested in two targeted deletions: a 2.9-kb deletion (Delta D MD Delta G) that removes the DMD and G-rich repeat and a 1.3-kb deletion (Delta G) removing only the latter. There are also four 21-bp GC-rich repetitive elements within the DMD that bind the insulator-associated CTCF (CCCTC-binding factor) protein and are implicated in mediating methylation-sensitive insulator activity. As three of the four repeats of the 2-kb DMD were deleted in the initial 1.6-kb Delta DMD allele, we analyzed a 3.8-kb targeted allele (Delta 3.8kb-5'H19), which deletes the entire DMD, to test the function of the fourth repeat. Comparative analysis of the 5' deletion alleles reveals that (i) the G-rich repeat element is dispensable for imprinting, (ii) the Delta DMD and Delta DMD Delta G alleles exhibit slightly more methylation upon paternal transmission, (iii) removal of the 5' CTCF site does not further perturb H19 and Igf2 imprinting, suggesting that one CTCF binding site is insufficient to generate insulator activity in vivo, (iv) the DMD sequence is required for full activation of H19 and Igf2, and (v) deletion of the DMD disrupts H19 and Igf2 expression in a tissue-specific manner. PMID- 11909941 TI - Histone H1 represses estrogen receptor alpha transcriptional activity by selectively inhibiting receptor-mediated transcription initiation. AB - Chromatin is the physiological template for many nuclear processes in eukaryotes, including transcription by RNA polymerase II. In vivo, chromatin is assembled from genomic DNA, core histones, linker histones such as histone H1, and nonhistone chromatin-associated proteins. Histone H1 is thought to act as a general repressor of transcription by promoting the compaction of chromatin into higher-order structures. We have used a biochemical approach, including an in vitro chromatin assembly and transcription system, to examine the effects of histone H1 on estrogen receptor alpha (ER alpha)-mediated transcription with chromatin templates. We show that histone H1 acts as a potent repressor of ligand and coactivator-regulated transcription by ER alpha. Histone H1 exerts its repressive effect without inhibiting the sequence-specific binding of ER alpha to chromatin or the overall extent of targeted acetylation of nucleosomal histones by the coactivator p300. Instead, histone H1 acts by blocking a specific step in the ER alpha-dependent transcription process, namely, transcription initiation, without affecting transcription reinitiation. Together, our data indicate that histone H1 acts selectively to reduce the overall level of productive transcription initiation by restricting promoter accessibility and preventing the ER alpha-dependent formation of a stable transcription pre-initiation complex. PMID- 11909942 TI - Enhanced endotoxin sensitivity in fps/fes-null mice with minimal defects in hematopoietic homeostasis. AB - The fps/fes proto-oncogene encodes a cytoplasmic protein tyrosine kinase implicated in growth factor and cytokine receptor signaling and thought to be essential for the survival and terminal differentiation of myeloid progenitors. Fps/Fes-null mice were healthy and fertile, displayed slightly reduced numbers of bone marrow myeloid progenitors and circulating mature myeloid cells, and were more sensitive to lipopolysaccharide (LPS). These phenotypes were rescued using a fps/fes transgene. This confirmed that Fps/Fes is involved in, but not required for, myelopoiesis and that it plays a role in regulating the innate immune response. Bone marrow-derived Fps/Fes-null macrophages showed no defects in granulocyte-macrophage colony-stimulating factor-, interleukin 6 (IL-6)-, or IL-3 induced activation of signal transducer and activator of transcription 3 (Stat3) and Stat5A or LPS-induced degradation of I kappa B or activation of p38, Jnk, Erk, or Akt. PMID- 11909943 TI - Critical but distinct roles for the pleckstrin homology and cysteine-rich domains as positive modulators of Vav2 signaling and transformation. AB - Vav2, like all Dbl family proteins, possesses tandem Dbl homology (DH) and pleckstrin homology (PH) domains and functions as a guanine nucleotide exchange factor for Rho family GTPases. Whereas the PH domain is a critical positive regulator of DH domain function for a majority of Dbl family proteins, the PH domains of the related Vav and Vav3 proteins are dispensable for DH domain activity. Instead, Vav proteins contain a cysteine-rich domain (CRD) critical for DH domain function. We evaluated the contribution of the PH domain and the CRD to Vav2 guanine nucleotide exchange, signaling, and transforming activity. Unexpectedly, we found that mutations of the PH domain impaired Vav2 signaling, transforming activity, and membrane association. However, these mutations do not influence exchange activity on Rac and only slightly affect exchange on RhoA and Cdc42. We also found that the CRD was critical for the exchange activity in vitro and contributed to Vav2 membrane localization. Finally, we found that phosphoinositol 3-kinase activation synergistically enhanced Vav2 transforming and signaling activity by stimulating exchange activity but not membrane association. In conclusion, the PH domain and CRD are mechanistically distinct, positive modulators of Vav2 DH domain function in vivo. PMID- 11909944 TI - Targeted disruption of Ras-Grf2 shows its dispensability for mouse growth and development. AB - The mammalian Grf1 and Grf2 proteins are Ras guanine nucleotide exchange factors (GEFs) sharing a high degree of structural homology, as well as an elevated expression level in central nervous system tissues. Such similarities raise questions concerning the specificity and/or redundancy at the functional level between the two Grf proteins. grf1-null mutant mice have been recently described which showed phenotypic growth reduction and long-term memory loss. To gain insight into the in vivo function of Grf2, we disrupted its catalytic CDC25-H domain by means of gene targeting. Breeding among grf2(+/-) animals gave rise to viable grf2(-/-) adult animals with a normal Mendelian pattern, suggesting that Grf2 is not essential for embryonic and adult mouse development. In contrast to Grf1-null mice, analysis of grf2(-/-) litters showed similar size and weight as their heterozygous or wild-type grf2 counterparts. Furthermore, adult grf2(-/-) animals reached sexual maturity at the same age as their wild-type littermates and showed similar fertility levels. No specific pathology was observed in adult Grf2-null animals, and histopathological studies showed no observable differences between null mutant and wild-type Grf2 mice. These results indicate that grf2 is dispensable for mouse growth, development, and fertility. Furthermore, analysis of double grf1/grf2 null animals did not show any observable phenotypic difference with single grf1(-/-) animals, further indicating a lack of functional overlapping between the two otherwise highly homologous Grf1 and Grf2 proteins. PMID- 11909945 TI - BP1, a homeodomain-containing isoform of DLX4, represses the beta-globin gene. AB - In earlier studies we identified a putative repressor of the human beta-globin gene, termed beta protein 1 (BP1), which binds to two silencer DNA sequences upstream of the adult human beta-globin gene and to a negative control region upstream of the adult delta-globin gene. Further studies demonstrated an inverse correlation between the binding affinity of the BP1 protein for the distal beta globin silencer sequence and the severity of sickle cell anemia, suggesting a possible role for BP1 in determining the production of hemoglobin S. We have now cloned a cDNA expressing the BP1 protein. Sequencing revealed that BP1 is a member of the homeobox gene family and belongs to the subfamily called Distal less (DLX), genes important in early development. Further analysis showed that BP1 is an isoform of DLX4. BP1 protein has repressor function towards the beta globin promoter, acting through the two beta-globin DNA silencers, demonstrated in transient transfection assays. Strong BP1 expression is restricted to placenta and kidney tissue, with no expression in 48 other human tissues. BP1 exhibits regulated expression in the human erythroid cell line MB-02, where its expression decreases upon induction of the beta-globin gene. BP1 is thus the first member of the DLX family with known DNA binding sites and a function in globin gene regulation. PMID- 11909946 TI - The response of c-jun/AP-1 to chronic hypoxia is hypoxia-inducible factor 1 alpha dependent. AB - Hypoxia (low-oxygen tension) is an important physiological stress that influences responses to a wide range of pathologies, including stroke, infarction, and tumorigenesis. Prolonged or chronic hypoxia stimulates expression of the stress inducible transcription factor gene c-jun and transient activation of protein kinase and phosphatase activities that regulate c-Jun/AP-1 activity. Here we describe evidence obtained by using wild-type and HIF-1 alpha nullizygous mouse embryonic fibroblasts (mEFs) that the induction of c-jun mRNA expression and c Jun phosphorylation by prolonged hypoxia are completely dependent on the presence of the oxygen-regulated transcription factor hypoxia-inducible factor 1 alpha (HIF-1 alpha). In contrast, transient hypoxia induced c-jun expression in both types of mEFs, showing that the early or rapid induction of this gene is independent of HIF-1 alpha. These findings indicate that the c-jun gene has a biphasic response to hypoxia consisting of inductions that depend on the degree or duration of exposure. To more completely define the relationship between prolonged hypoxia and c-Jun phosphorylation, we used mEFs from mice containing inactivating mutations of critical phosphorylation sites in the c-Jun N-terminal region (serines 63 and 73 or threonines 91 and 93). Exposure of these mEFs to prolonged hypoxia demonstrated an absolute requirement for N-terminal sites for HIF-1 alpha-dependent phosphorylation of c-Jun. Taken together, these findings suggest that c-Jun/AP-1 and HIF-1 cooperate to regulate gene expression in pathophysiological microenvironments. PMID- 11909947 TI - The direct recruitment of BLNK to immunoglobulin alpha couples the B-cell antigen receptor to distal signaling pathways. AB - Following B-cell antigen receptor (BCR) ligation, the cytoplasmic domains of immunoglobulin alpha (Ig alpha) and Ig beta recruit Syk to initiate signaling cascades. The coupling of Syk to several distal substrates requires linker protein BLNK. However, the mechanism by which BLNK is recruited to the BCR is unknown. Using chimeric receptors with wild-type and mutant Ig alpha cytoplasmic tails we show that the non-immunoreceptor tyrosine-based activation motif (ITAM) tyrosines, Y176 and Y204, are required to activate BLNK-dependent pathways. Subsequent analysis demonstrated that BLNK bound directly to phospho-Y204 and that fusing BLNK to mutated Ig alpha reconstituted downstream signaling events. Moreover, ligation of the endogenous BCR induced Y204 phosphorylation and BLNK recruitment. These data demonstrate that the non-ITAM tyrosines of Ig alpha couple Syk activation to BLNK-dependent pathways. PMID- 11909948 TI - Tumor necrosis factor receptor 1 is an ATPase regulated by silencer of death domain. AB - Self-aggregation of tumor necrosis factor receptor type 1 (TNFR1) induces spontaneous downstream signaling and results in cell death. It has been suggested that silencer of death domain (SODD) binds TNFR1 monomers to prevent self aggregation. We found that SODD binds through its BAG domain to the ATPase domain of Hsp70. We also determined that SODD binds through its BAG domain to TNFR1. ATP, but not nonhydrolyzable ATP-gamma S, regulates the SODD binding by Hsp70 or TNFR1. ATP binding by TNFR1 was abolished when a point mutation was introduced into a phosphate-binding loop motif characteristic of ATP-binding proteins, suggesting that TNFR1 functions as an ATPase. Furthermore, TNFR1 was present in aggregates in ATP-depleted cells and SODD disassembled aggregates in vitro only in the presence of ATP. These data suggest that SODD functions as a cofactor analogous to the nucleotide exchange factor BAG-1, which modulates the ATPase cycle of Hsp70 proteins. We propose a new model in which a nucleotide-dependent conformational change in TNFR1 has a key role in regulating TNF signaling. PMID- 11909949 TI - Kap121p-mediated nuclear import is required for mating and cellular differentiation in yeast. AB - To further our understanding of how the nucleocytoplasmic transport machinery interfaces with its cargoes and how this affects cellular physiology, we investigated the molecular mechanisms of phenotypes associated with mutations in karyopherin Kap121p. Two previously unreported phenotypes of kap121 cells were observed: defects in mating and in the transition from the normal yeast form to the pseudohyphal, invasive form. In parallel, we searched for Kap121p cargoes by using Kap121p as a probe in overlay assays of yeast nuclear proteins. One of the major interacting proteins identified by this procedure was Ste12p, a transcription factor central to both the mating response and the pseudohyphal transition. We therefore investigated whether defects in these differentiation processes were due to an inability to import Ste12p. Both immunopurification and in vitro binding studies demonstrated that Ste12p interacted specifically with Kap121p in a Ran-GTP-sensitive manner and that Ste12p was mislocalized to the cytoplasm by inactivation of Kap121p in a temperature-sensitive mutant. The Kap121p-specific nuclear localization signal (NLS) of Ste12p was determined to reside within a C-terminal region of Ste12p. Furthermore, by overexpression of STE12 or expression of a STE12-cNLS fusion in kap121 cells, the invasive-growth defect and the mating defect were both suppressed. Together these data demonstrate that Ste12p is imported into nuclei by Kap121p and that mating and differentiation defects associated with kap121 mutants are primarily attributable to the mislocalization of Ste12p. PMID- 11909951 TI - Novel G-protein complex whose requirement is linked to the translational status of the cell. AB - G proteins, which bind and hydrolyze GTP, are involved in regulating a variety of critical cellular processes, including the process of protein synthesis. Many members of the subfamily of elongation factor class G proteins interact with the ribosome and function to regulate discrete steps during the process of protein synthesis. Despite sequence similarity to factors involved in translation, a role for the yeast Hbs1 protein has not been defined. In this work we have identified a genetic relationship between genes encoding components of the translational apparatus and HBS1. HBS1, while not essential for viability, is important for efficient growth and protein synthesis under conditions of limiting translation initiation. The identification of an Hbs1p-interacting factor, Dom34p, which shares a similar genetic relationship with components of the translational apparatus, suggests that Hbs1p and Dom34p may function as part of a complex that facilitates gene expression. Dom34p contains an RNA binding motif present in several ribosomal proteins and factors that regulate translation of specific mRNAs. Thus, Hbs1p and Dom34p may function together to help directly or indirectly facilitate the expression either of specific mRNAs or under certain cellular conditions. PMID- 11909950 TI - Central role for the XRCC1 BRCT I domain in mammalian DNA single-strand break repair. AB - The DNA single-strand break repair (SSBR) protein XRCC1 is required for genetic stability and for embryonic viability. XRCC1 possesses two BRCA1 carboxyl terminal (BRCT) protein interaction domains, denoted BRCT I and II. BRCT II is required for SSBR during G(1) but is dispensable for this process during S/G(2) and consequently for cell survival following DNA alkylation. Little is known about BRCT I, but this domain has attracted considerable interest because it is the site of a genetic polymorphism that epidemiological studies have associated with altered cancer risk. We report that the BRCT I domain comprises the evolutionarily conserved core of XRCC1 and that this domain is required for efficient SSBR during both G(1) and S/G(2) cell cycle phases and for cell survival following treatment with methyl methanesulfonate. However, the naturally occurring human polymorphism in BRCT I supported XRCC1-dependent SSBR and cell survival after DNA alkylation equally well. We conclude that while the BRCT I domain is critical for XRCC1 to maintain genetic integrity and cell survival, the polymorphism does not impact significantly on this function and therefore is unlikely to impact significantly on susceptibility to cancer. PMID- 11909952 TI - Autoinhibitory regulation of p73 by Delta Np73 to modulate cell survival and death through a p73-specific target element within the Delta Np73 promoter. AB - p73 is a p53-related tumor suppressor but is also induced by oncogene products such as E2F-1, raising a question as to whether p73 is a tumor suppressor gene or oncogene. Unlike p53, p73 has several variants, including Delta Np73, which lacks the NH(2)-terminal transactivation domain. Although, in developing neurons, Delta Np73 is expressed abundantly and seems to inhibit the proapoptotic function of p53, the role of p73 and Delta Np73 and their regulatory mechanism in cell growth and differentiation are poorly understood. Here we report that p73, but not p53, directly activates the transcription of endogenous Delta Np73 by binding to the p73-specific target element located at positions -76 to -57 within the Delta Np73 promoter region. The activation of Delta Np73 promoter by p63 was marginal. Delta Np73 was associated with p73 alpha, p73 beta, and p53, as demonstrated by immunoprecipitation assays, and inhibited their transactivation activities when we used reporters of Mdm2, Bax, or Delta Np73 itself in SAOS-2 cells. Furthermore, induction or overexpression of Delta Np73 promoted cell survival by competing with p53 and p73 itself. Thus, our results suggest that the negative feedback regulation of p73 by its target Delta Np73 is a novel autoregulatory system for modulating cell survival and death. PMID- 11909953 TI - Cripto-1 activates nodal- and ALK4-dependent and -independent signaling pathways in mammary epithelial Cells. AB - Cripto-1 (CR-1), an epidermal growth factor-CFC (EGF-CFC) family member, has a demonstrated role in embryogenesis and mammary gland development and is overexpressed in several human tumors. Recently, EGF-CFC proteins were implicated as essential signaling cofactors for Nodal, a transforming growth factor beta family member whose expression has previously been defined as embryo specific. To identify a receptor for CR-1, a human brain cDNA phage display library was screened using CR-1 protein as bait. Phage inserts with identity to ALK4, a type I serine/threonine kinase receptor for Activin, were identified. CR-1 binds to cell surface ALK4 expressed on mammalian epithelial cells in fluorescence activated cell sorter analysis, as well as by coimmunoprecipitation. Nodal is coexpressed with mouse Cr-1 in the mammary gland, and CR-1 can phosphorylate the transcription factor Smad-2 in EpH-4 mammary epithelial cells only in the presence of Nodal and ALK4. In contrast, CR-1 stimulation of mitogen-activated protein kinase and AKT in these cells is independent of Nodal and ALK4, suggesting that CR-1 may modulate different signaling pathways to mediate its different functional roles. PMID- 11909954 TI - Scaffold/matrix attachment region elements interact with a p300-scaffold attachment factor A complex and are bound by acetylated nucleosomes. AB - The transcriptional coactivator p300 regulates transcription by binding to proteins involved in transcription and by acetylating histones and other proteins. These transcriptional effects are mainly at promoter and enhancer elements. Regulation of transcription also occurs through scaffold/matrix attachment regions (S/MARs), the chromatin regions that bind the nuclear matrix. Here we show that p300 binds to the S/MAR binding protein scaffold attachment factor A (SAF-A), a major constituent of the nuclear matrix. Using chromatin immunoprecipitations, we established that both p300 and SAF-A bind to S/MAR elements in the transiently silent topoisomerase I gene prior to its activation at G(1) during cell cycle. This binding is accompanied by local acetylation of nucleosomes, suggesting that p300-SAF-A interactions at S/MAR elements of nontranscribed genes might poise these genes for transcription. PMID- 11909956 TI - Inducer-specific enhanceosome formation controls tumor necrosis factor alpha gene expression in T lymphocytes. AB - We present evidence that the inducer-specific regulation of the human tumor necrosis factor alpha (TNF-alpha) gene in T cells involves the assembly of distinct higher-order transcription enhancer complexes (enhanceosomes), which is dependent upon inducer-specific helical phasing relationships between transcription factor binding sites. While ATF-2, c-Jun, and the coactivator proteins CBP/p300 play a central role in TNF-alpha gene activation stimulated by virus infection or intracellular calcium flux, different sets of activators including NFATp, Sp1, and Ets/Elk are recruited to a shared set of transcription factor binding sites depending upon the particular stimulus. Thus, these studies demonstrate that the inducer-specific assembly of unique enhanceosomes is a general mechanism by which a single gene is controlled in response to different extracellular stimuli. PMID- 11909957 TI - Direct channeling of retinoic acid between cellular retinoic acid-binding protein II and retinoic acid receptor sensitizes mammary carcinoma cells to retinoic acid induced growth arrest. AB - Cellular retinoic acid-binding protein II (CRABP-II) is an intracellular lipid binding protein that associates with retinoic acid with a subnanomolar affinity. We previously showed that CRABP-II enhances the transcriptional activity of the nuclear receptor with which it shares a common ligand, namely, the retinoic acid receptor (RAR), and we suggested that it may act by delivering retinoic acid to this receptor. Here, the mechanisms underlying the effects of CRABP-II on the transcriptional activity of RAR and the functional consequences of these effects were studied. We show that CRABP-II, a predominantly cytosolic protein, massively undergoes nuclear localization upon binding of retinoic acid; that it interacts with RAR in a ligand-dependent fashion; and that, in the presence of retinoic acid, the CRABP-II-RAR complex is a short-lived intermediate. The data establish that potentiation of the transcriptional activity of RAR stems directly from the ability of CRABP-II to channel retinoic acid to the receptor. We demonstrate further that overexpression of CRABP-II in MCF-7 mammary carcinoma cells dramatically enhances their sensitivity to retinoic acid-induced growth inhibition. Conversely, diminished expression of CRABP-II renders these cells retinoic acid resistant. Taken together, the data unequivocally establish the function of CRABP-II in modulating the RAR-mediated biological activities of retinoic acid. PMID- 11909955 TI - Conditional disruption of the peroxisome proliferator-activated receptor gamma gene in mice results in lowered expression of ABCA1, ABCG1, and apoE in macrophages and reduced cholesterol efflux. AB - Disruption of the peroxisome proliferator-activated receptor gamma (PPAR gamma) gene causes embryonic lethality due to placental dysfunction. To circumvent this, a PPAR gamma conditional gene knockout mouse was produced by using the Cre-loxP system. The targeted allele, containing loxP sites flanking exon 2 of the PPAR gamma gene, was crossed into a transgenic mouse line expressing Cre recombinase under the control of the alpha/beta interferon-inducible (MX) promoter. Induction of the MX promoter by pIpC resulted in nearly complete deletion of the targeted exon, a corresponding loss of full-length PPAR gamma mRNA transcript and protein, and marked reductions in basal and troglitazone-stimulated expression of the genes encoding lipoprotein lipase, CD36, LXR alpha, and ABCG1 in thioglycolate elicited peritoneal macrophages. Reductions in the basal levels of apolipoprotein E (apoE) mRNA in macrophages and apoE protein in total plasma and high-density lipoprotein (HDL) were also observed in pIpC-treated PPAR gamma-MXCre(+) mice. Basal cholesterol efflux from cholesterol-loaded macrophages to HDL was significantly reduced after disruption of the PPAR gamma gene. Troglitazone selectively inhibited ABCA1 expression (while rosiglitazone, ciglitazone, and pioglitazone had little effect) and cholesterol efflux in both PPAR gamma deficient and control macrophages, indicating that this drug can exert paradoxical effects on cholesterol homeostasis that are independent of PPAR gamma. Together, these data indicate that PPAR gamma plays a critical role in the regulation of cholesterol homeostasis by controlling the expression of a network of genes that mediate cholesterol efflux from cells and its transport in plasma. PMID- 11909958 TI - New insights into the pleiotropic drug resistance network from genome-wide characterization of the YRR1 transcription factor regulation system. AB - Yrr1p is a recently described Zn(2)Cys(6) transcription factor involved in the pleiotropic drug resistance (PDR) phenomenon. It is controlled in a Pdr1p dependent manner and is autoregulated. We describe here a new genome-wide approach to characterization of the set of genes directly regulated by Yrr1p. We found that the time-course production of an artificial chimera protein containing the DNA-binding domain of Yrr1p activated the 15 genes that are also up-regulated by a gain-of-function mutant of Yrr1p. Gel mobility shift assays showed that the promoters of the genes AZR1, FLR1, SNG1, YLL056C, YLR346C, and YPL088W interacted with Yrr1p. The putative consensus Yrr1p binding site deduced from these experiments, (T/A)CCG(C/T)(G/T)(G/T)(A/T)(A/T), is strikingly similar to the PDR element binding site sequence recognized by Pdr1p and Pdr3p. The minor differences between these sequences are consistent with Yrr1p and Pdr1p and Pdr3p having different sets of target genes. According to these data, some target genes are directly regulated by Pdr1p and Pdr3p or by Yrr1p, whereas some genes are indirectly regulated by the activation of Yrr1p. Some genes, such as YOR1, SNQ2, and FLR1, are clearly directly controlled by both classes of transcription factor, suggesting an important role for the corresponding membrane proteins. PMID- 11909959 TI - Nir2, a novel regulator of cell morphogenesis. AB - Cell morphogenesis requires dynamic reorganization of the actin cytoskeleton, a process that is tightly regulated by the Rho family of small GTPases. These GTPases act as molecular switches by shuttling between their inactive GDP-bound and active GTP-bound forms. Here we show that Nir2, a novel protein related to Drosophila retinal degeneration B (RdgB), markedly affects cell morphology through a novel Rho-inhibitory domain (Rid) which resides in its N-terminal region. Rid exhibits sequence homology with the Rho-binding site of formin homology (FH) proteins and leads to an apparent loss of F-actin staining when ectopically expressed in mammalian cells. We also show that Rid inhibits Rho mediated stress fiber formation and lysophosphatidic acid-induced RhoA activation. Biochemical studies demonstrated that Nir2, via Rid, preferentially binds to the inactive GDP-bound form of the small GTPase Rho. Microinjection of antibodies against Nir2 into neuronal cells markedly attenuates neurite extension, whereas overexpression of Nir2 in these cells attenuates Rho-mediated neurite retraction. These results implicate Nir2 as a novel regulator of the small GTPase Rho in actin cytoskeleton reorganization and cell morphogenesis. PMID- 11909960 TI - Mutant mouse models reveal the relative roles of E2F1 and E2F3 in vivo. AB - The E2F1, -2, and -3 transcription factors are key downstream targets of the retinoblastoma protein (pRB) tumor suppressor that drive expression of proliferation-associated genes. Here we use mutant mouse strains to investigate E2F3's role in vivo. We show that E2F3 is essential for embryonic viability in the pure 129/Sv background but the presence of C57BL/6 alleles yields some adult survivors. Although growth retarded, surviving E2f3(-/-) animals are initially healthy. However, they die prematurely, exhibiting no obvious tumor phenotype but with the typical signs of congestive heart failure. The defects are completely distinct from those arising in E2f1 mutant mice (S. J. Field et al., Cell 85:549 561; 1996; L. Yamasaki et al., Cell 85:537-548, 1996), supporting the prevailing view that these E2Fs must have some unique biological functions in vivo. To test this model, we examined the phenotypes of E2f1 E2f3 compound mutant mice. Almost all of the developmental and age-related defects arising in the individual E2f1 or E2f3 mice were exacerbated by the mutation of the other E2f. Thus, E2F1 and E2F3 appear to play critical, overlapping roles in the development and maintenance of a variety of tissues. Importantly, this study did identify one major difference in the properties of E2F1 and E2F3: either alone or in combination with E2F1 loss, E2f3 mutation did not increase the incidence of tumor formation. These data strongly suggest that tumor suppression is a specific property of E2F1 and not E2F3. PMID- 11909961 TI - SKAP55 coupled with CD45 positively regulates T-cell receptor-mediated gene transcription. AB - CD45 plays a critical role in T-cell receptor (TCR)-mediated signaling. In a yeast two-hybrid screen, SKAP55, the Src kinase-associated phosphoprotein of unknown function, was found as a substrate which associated with CD45 in vivo. Mutational analysis demonstrated the pivotal role of Tyr-232 in SKAP55 in the association with CD45. In Jurkat cells, anti-CD3 antibody stimulation promoted SKAP55 tyrosine phosphorylation and translocation from the cytoplasm to the membrane. Overexpression of SKAP55 in these cells induced transcriptional activation of the IL-2 promoter, while mutant SKAP55-Y232F totally suppressed the promoter activity. Furthermore, overexpression of SKAP55-Y232F also caused the tyrosine hyperphosphorylation of Fyn with a decreased kinase activity. Thus, SKAP55 is an essential adapter to couple CD45 with the Src family kinases for dephosphorylation and, thus, positively regulates TCR signaling. PMID- 11909962 TI - Sp100 interacts with ETS-1 and stimulates its transcriptional activity. AB - The cell nucleus is highly organized into distinct domains that spatially separate physiological processes. One of these domains, the Sp100-promyelocytic leukemia protein nuclear body (NB), is implicated in pathological processes, such as cancer and viral infection, yet its functions remain poorly understood. We show here that Sp100 interacts physically and functionally with ETS-1 and that NB morphology is affected by ETS-1. ETS-1 is a member of the ets family of transcription factors, which are key mediators of physiological and pathological processes. We have found that Sp100 interacts with two regions of ETS-1 (domains A+B and D+E+F). ETS-1 alters NBs while remaining localized throughout the nucleus, apparently by recruitment of the core component Sp100 away from the NBs. Sp100 strongly increases ETS-1 activation of natural and ets-focused promoters, through a mechanism involving the activation (C) domain of ETS-1 in addition to the interaction domains. Sp100 acts as a novel coactivator that potentiates the activator function of ETS-1. Our results provide an important new connection between nuclear structures and an important regulator of gene expression. PMID- 11909963 TI - Scatter factor/hepatocyte growth factor stimulation of glioblastoma cell cycle progression through G(1) is c-Myc dependent and independent of p27 suppression, Cdk2 activation, or E2F1-dependent transcription. AB - Scatter factor/hepatocyte growth factor (SF/HGF) expression has been linked to malignant progression in glial neoplasms. Using two glioma cell lines, U373MG and SNB-19, we have demonstrated that SF/HGF stimulation allows cells to escape G(1)/G(0) arrest induced by contact inhibition or serum withdrawal. SF/HGF induced effects on two mechanisms of cell cycle regulation: suppression of the cyclin-dependent kinase inhibitor p27 and induction of the transcription factor c Myc. Regulation of p27 by SF/HGF was posttranslational and is associated with p27 nuclear export. Transient transfections of U373MG and SNB-19 with wild-type p27 and a degradation-resistant p27T187A mutant were insufficient to induce cell cycle arrest, and SF/HGF downregulation of p27 was not necessary for cell cycle reentry. Analysis of Cdk2 kinase activity and p27 binding to cyclin E complexes in the presence of exogenous wild-type p27 or p27T187A demonstrated that Cdk2 activity was not necessary for SF/HGF-mediated G(1)/S transition. Similarly, overexpression of dominant-negative forms of Cdk2 did not block SF/HGF-triggered cell cycle progression. In contrast, SF/HGF transcriptionally upregulated c-Myc, and overexpression of c-Myc was able to prevent G(1)/G(0) arrest in the absence of SF/HGF. Transient overexpression of MadMyc, a dominant-negative chimera for c Myc, caused G(1)/G(0) arrest in logarithmically growing cells and blocked SF/HGF mediated G(1)/S transition. c-Myc did not exert its effects through p27 downregulation in these cell lines. SF/HGF induced E2F1-dependent transcription, the inhibition of which did not block SF/HGF-induced cell cycle progression. We conclude that SF/HGF prevents G(1)/G(0) arrest in glioma cell lines by a c-myc dependent mechanism that is independent of p27, Cdk2, or E2F1. PMID- 11909964 TI - Activation of m-calpain (calpain II) by epidermal growth factor is limited by protein kinase A phosphorylation of m-calpain. AB - We have shown previously that the ELR-negative CXC chemokines interferon inducible protein 10, monokine induced by gamma interferon, and platelet factor 4 inhibit epidermal growth factor (EGF)-induced m-calpain activation and thereby EGF-induced fibroblast cell motility (H. Shiraha, A. Glading, K. Gupta, and A. Wells, J. Cell Biol. 146:243-253, 1999). However, how this cross attenuation could be accomplished remained unknown since the molecular basis of physiological m-calpain regulation is unknown. As the initial operative attenuation signal from the CXCR3 receptor was cyclic AMP (cAMP), we verified that this second messenger blocked EGF-induced motility of fibroblasts (55% +/- 4.5% inhibition) by preventing rear release during active locomotion. EGF-induced calpain activation was inhibited by cAMP activation of protein kinase A (PKA), as the PKA inhibitors H-89 and Rp-8Br-cAMPS abrogated cAMP inhibition of both motility and calpain activation. We hypothesized that PKA might negatively modulate m-calpain in an unexpected manner by directly phosphorylating m-calpain. A mutant human large subunit of m-calpain was genetically engineered to negate a putative PKA consensus sequence in the regulatory domain III (ST369/370AA) and was expressed in NR6WT mouse fibroblasts to represent about 30% of total m-calpain in these cells. This construct was not phosphorylated by PKA in vitro while a wild-type construct was, providing proof of the principle that m-calpain can be directly phosphorylated by PKA at this site. cAMP suppressed EGF-induced calpain activity of cells overexpressing a control wild-type human m-calpain (83% +/- 3.7% inhibition) but only marginally suppressed that of cells expressing the PKA resistant mutant human m-calpain (25% +/- 5.5% inhibition). The EGF-induced motility of the cells expressing the PKA-resistant mutant also was not inhibited by cAMP. Structural modeling revealed that new constraints resulting from phosphorylation at serine 369 would restrict domain movement and help "freeze" m calpain in an inactive state. These data point to a novel mechanism of negative control of calpain activation, direct phosphorylation by PKA. PMID- 11909965 TI - Fission yeast Mad3p is required for Mad2p to inhibit the anaphase-promoting complex and localizes to kinetochores in a Bub1p-, Bub3p-, and Mph1p-dependent manner. AB - The spindle checkpoint delays the metaphase-to-anaphase transition in response to spindle and kinetochore defects. Genetic screens in budding yeast identified the Mad and Bub proteins as key components of this conserved regulatory pathway. Here we present the fission yeast homologue of Mad3p. Cells devoid of mad3(+) are unable to arrest their cell cycle in the presence of microtubule defects. Mad3p coimmunoprecipitates Bub3p, Mad2p, and the spindle checkpoint effector Slp1/Cdc20p. We demonstrate that Mad3p function is required for the overexpression of Mad2p to result in a metaphase arrest. Mad1p, Bub1p, and Bub3p are not required for this arrest. Thus, Mad3p appears to have a crucial role in transducing the inhibitory "wait anaphase" signal to the anaphase-promoting complex (APC). Mad3-green fluorescent protein (GFP) is recruited to unattached kinetochores early in mitosis and accumulates there upon prolonged checkpoint activation. For the first time, we have systematically studied the dependency of Mad3/BubR1 protein recruitment to kinetochores. We find Mad3-GFP kinetochore localization to be dependent upon Bub1p, Bub3p, and the Mph1p kinase, but not upon Mad1p or Mad2p. We discuss the implications of these findings in the context of our current understanding of spindle checkpoint function. PMID- 11909966 TI - Identification of mammalian Sds3 as an integral component of the Sin3/histone deacetylase corepressor complex. AB - Silencing of gene transcription involves local chromatin modification achieved through the local recruitment of large multiprotein complexes containing histone deacetylase (HDAC) activity. The mammalian corepressors mSin3A and mSin3B have been shown to play a key role in this process by tethering HDACs 1 and 2 to promoter-bound transcription factors. Similar mechanisms appear to be operative in yeast, in which epistasis experiments have established that the mSin3 and HDAC orthologs (SIN3 and RPD3), along with a novel protein, SDS3, function in the same repressor pathway. Here, we report the identification of a component of the mSin3 HDAC complex that bears homology to yeast SDS3, physically associates with mSin3 proteins in vivo, represses transcription in a manner that is partially dependent on HDAC activity, and enables HDAC1 catalytic activity in vivo. That key physical and functional properties are also shared by yeast SDS3 underscores the central role of the Sin3-HDAC-Sds3 complex in eukaryotic cell biology, and the discovery of mSds3 in mammalian cells provides a new avenue for modulating the activity of this complex in human disease. PMID- 11909967 TI - Structural insight into the mechanisms of targeting and signaling of focal adhesion kinase. AB - Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase whose focal adhesion targeting (FAT) domain interacts with other focal adhesion molecules in integrin mediated signaling. Localization of activated FAK to focal adhesions is indispensable for its function. Here we describe a solution structure of the FAT domain bound to a peptide derived from paxillin, a FAK-binding partner. The FAT domain is composed of four helices that form a "right-turn" elongated bundle; the globular fold is mainly maintained by hydrophobic interactions. The bound peptide further stabilizes the structure. Certain signaling events such as phosphorylation and molecule interplay may induce opening of the helix bundle. Such conformational change is proposed to precede departure of FAK from focal adhesions, which starts focal adhesion turnover. PMID- 11909968 TI - Reduced sperm count and normal fertility in male mice with targeted disruption of the ADP-ribosylation factor-like 4 (Arl4) gene. AB - The ADP-ribosylation factor-like protein 4 (ARL4) is a 22-kDa GTP-binding protein which is abundant in testes of pubertal and adult rodents but absent in testes from prepubertal animals. During testis development, ARL4 expression starts at day 16 when the spermatogenesis proceeds to the late pachytene. In the adult testis, the ARL4 protein was detected in pre- and postmeiotic cells, spermatocytes, and spermatides, but not in spermatogonia and mature spermatozoa. Mouse Arl4-null mutants generated by targeted disruption of the Arl4 gene were viable and grew normally; male as well as female Arl4(-/-) mice were fertile. However, inactivation of the Arl4 gene resulted in a significant reduction of testis weight and sperm count by 30 and 60%, respectively, without reduction of litter size or frequency. It is suggested that the disruption of Arl4 produces a moderate retardation of germ cell development, possibly at the stage of meiosis. PMID- 11909969 TI - Hydrocephalus, situs inversus, chronic sinusitis, and male infertility in DNA polymerase lambda-deficient mice: possible implication for the pathogenesis of immotile cilia syndrome. AB - A growing number of DNA polymerases have been identified, although their physiological function and relation to human disease remain mostly unknown. DNA polymerase lambda (Pol lambda; also known as Pol beta2) has recently been identified as a member of the X family of DNA polymerases and shares 32% amino acid sequence identity with DNA Pol beta within the polymerase domain. With the use of homologous recombination, we generated Pol lambda(-/-) mice. Pol lambda(-/ ) mice develop hydrocephalus with marked dilation of the lateral ventricles and exhibit a high rate of mortality after birth, although embryonic development appears normal. Pol lambda(-/-) mice also show situs inversus totalis and chronic suppurative sinusitis. The surviving male, but not female, Pol lambda(-/-) mice are sterile as a result of spermatozoal immobility. Microinjection of sperm from male Pol lambda(-/-) mice into oocytes gives rise to normal offspring, suggesting that the meiotic process is not impaired. Ultrastructural analysis reveals that inner dynein arms of cilia from both the ependymal cell layer and respiratory epithelium are defective, which may underlie the pathogenesis of hydrocephalus, situs inversus totalis, chronic sinusitis, and male infertility. Sensitivity of Pol lambda(-/-) cells to various kinds of DNA damage is indistinguishable from that of Pol lambda(+/+) cells. Collectively, Pol lambda(-/-) mice may provide a useful model for clarifying the pathogenesis of immotile cilia syndrome. PMID- 11909970 TI - Stat1-vitamin D receptor interactions antagonize 1,25-dihydroxyvitamin D transcriptional activity and enhance stat1-mediated transcription. AB - The cytokine gamma interferon (IFN-gamma) and the calcitropic steroid hormone 1,25-dihydroxyvitamin D (1,25D) are activators of macrophage immune function. In sarcoidosis, tuberculosis, and several granulomatoses, IFN-gamma induces 1,25D synthesis by macrophages and inhibits 1,25D induction of 24-hydroxylase, a key enzyme in 1,25D inactivation, causing high levels of 1,25D in serum and hypercalcemia. This study delineates IFN-gamma-1,25D cross talk in human monocytes-macrophages. Nuclear accumulation of Stat1 and vitamin D receptor (VDR) by IFN-gamma and 1,25D promotes protein-protein interactions between Stat1 and the DNA binding domain of the VDR. This prevents VDR-retinoid X receptor (RXR) binding to the vitamin D-responsive element, thus diverting the VDR from its normal genomic target on the 24-hydroxylase promoter and antagonizing 1,25D-VDR transactivation of this gene. In contrast, 1,25D enhances IFN-gamma action. Stat1 VDR interactions, by preventing Stat1 deactivation by tyrosine dephosphorylation, cooperate with IFN-gamma/Stat1-induced transcription. This novel 1,25D-IFN-gamma cross talk explains the pathogenesis of abnormal 1,25D homeostasis in granulomatous processes and provides new insights into 1,25D immunomodulatory properties. PMID- 11909971 TI - Evidence that TAF-TATA box-binding protein interactions are required for activated transcription in mammalian cells. AB - Surfaces of human TATA box-binding protein (hsTBP) required for activated transcription in vivo were defined by constructing a library of surface residue substitution mutations and assaying them for their ability to support activated transcription in transient-transfection assays. In earlier work, three regions were identified where mutations inhibited activated transcription without interfering with TATA box DNA binding. One region is on the upstream surface of the N-terminal TBP repeat with respect to the direction of transcription and corresponds to the TBP surface that interacts with TFIIA. A second region on the stirrup of the C-terminal TBP repeat corresponds to the TFIIB-binding surface. Here we report that the third region where mutations inhibit activated transcription in mammalian cells, the convex surface of the N-terminal repeat, corresponds to a surface on TBP that interacts with hsTAF1, the major scaffold subunit of TFIID. Since mutations at the center of the hsTAF1-interacting region inhibit the ability of the protein to support activated transcription in vivo, these results are consistent with the conclusion that an interaction between hsTBP and TAF(II)s is required for activated transcription in mammalian cells. PMID- 11909972 TI - Akt/protein kinase B promotes organ growth in transgenic mice. AB - One of the least-understood areas in biology is the determination of the size of animals and their organs. In Drosophila, components of the insulin receptor phosphoinositide 3-kinase (PI3K) pathway determine body, organ, and cell size. Several biochemical studies have suggested that Akt/protein kinase B is one of the important downstream targets of PI3K. To examine the role of Akt in the regulation of organ size in mammals, we have generated and characterized transgenic mice expressing constitutively active Akt (caAkt) or kinase-deficient Akt (kdAkt) specifically in the heart. The heart weight of caAkt transgenic mice was increased 2.0-fold compared with that of nontransgenic mice. The increase in heart size was associated with a comparable increase in myocyte cell size in caAkt mice. The kdAkt mutant protein attenuated the constitutively active PI3K induced overgrowth of the heart, and the caAkt mutant protein circumvented cardiac growth retardation induced by a kinase-deficient PI3K mutant protein. Rapamycin attenuated caAkt-induced overgrowth of the heart, suggesting that the mammalian target of rapamycin (mTOR) or effectors of mTOR mediated caAkt-induced heart growth. In conclusion, Akt is sufficient to induce a marked increase in heart size and is likely to be one of the effectors of the PI3K pathway in mediating heart growth. PMID- 11909974 TI - Impaired cardiac contractility response to hemodynamic stress in S100A1-deficient mice. AB - Ca(2+) signaling plays a central role in cardiac contractility and adaptation to increased hemodynamic demand. We have generated mice with a targeted deletion of the S100A1 gene coding for the major cardiac isoform of the large multigenic S100 family of EF hand Ca(2+)-binding proteins. S100A1(-/-) mice have normal cardiac function under baseline conditions but have significantly reduced contraction rate and relaxation rate responses to beta-adrenergic stimulation that are associated with a reduced Ca(2+) sensitivity. In S100A1(-/-) mice, basal left ventricular contractility deteriorated following 3-week pressure overload by thoracic aorta constriction despite a normal adaptive hypertrophy. Surprisingly, heterozygotes also had an impaired response to acute beta-adrenergic stimulation but maintained normal contractility in response to chronic pressure overload that coincided with S100A1 upregulation to wild-type levels. In contrast to other genetic models with impaired cardiac contractility, loss of S100A1 did not lead to cardiac hypertrophy or dilation in aged mice. The data demonstrate that high S100A1 protein levels are essential for the cardiac reserve and adaptation to acute and chronic hemodynamic stress in vivo. PMID- 11909973 TI - HMG2 interacts with the nucleosome assembly protein SET and is a target of the cytotoxic T-lymphocyte protease granzyme A. AB - The cytotoxic T-lymphocyte protease granzyme A induces caspase-independent cell death in which DNA single-stranded nicking is observed instead of oligonucleosomal fragmentation. A 270- to 420-kDa endoplasmic reticulum associated complex (SET complex) containing the nucleosome assembly protein SET, the tumor suppressor pp32, and the base excision repair enzyme APE can induce single-stranded DNA damage in isolated nuclei in a granzyme A-dependent manner. The normal functions of the SET complex are unknown, but the functions of its components suggest that it is involved in activating transcription and DNA repair. We now find that the SET complex contains DNA binding and bending activities mediated by the chromatin-associated protein HMG2. HMG2 facilitates assembly of nucleoprotein higher-order structures by bending and looping DNA or by stabilizing underwound DNA. HMG2 is in the SET complex and coprecipitates with SET. By confocal microscopy, it is observed that cytoplasmic HMG2 colocalizes with SET in association with the endoplasmic reticulum, but most nuclear HMG2 is unassociated with SET. This physical association suggests that HMG2 may facilitate the nucleosome assembly, transcriptional activation, and DNA repair functions of SET and/or APE. HMG2, like SET and APE, is a physiologically relevant granzyme A substrate in targeted cells. HMG1, however, is not a substrate. Granzyme A cleavage after Lys65 in the midst of HMG box A destroys HMG2-mediated DNA binding and bending functions. Granzyme A cleavage and functional disruption of key nuclear substrates, including HMG2, SET, APE, lamins, and histones, are likely to cripple the cellular repair response to promote cell death in this novel caspase-independent death pathway. PMID- 11909975 TI - Activated Notch4 inhibits angiogenesis: role of beta 1-integrin activation. AB - Notch4 is a member of the Notch family of transmembrane receptors that is expressed primarily on endothelial cells. Activation of Notch in various cell systems has been shown to regulate cell fate decisions. The sprouting of endothelial cells from microvessels, or angiogenesis, involves the modulation of the endothelial cell phenotype. Based on the function of other Notch family members and the expression pattern of Notch4, we postulated that Notch4 activation would modulate angiogenesis. Using an in vitro endothelial-sprouting assay, we show that expression of constitutively active Notch4 in human dermal microvascular endothelial cells (HMEC-1) inhibits endothelial sprouting. We also show that activated Notch4 inhibits vascular endothelial growth factor (VEGF) induced angiogenesis in the chick chorioallantoic membrane in vivo. Activated Notch4 does not inhibit HMEC-1 proliferation or migration through fibrinogen. However, migration through collagen is inhibited. Our data show that Notch4 cells exhibit increased beta1-integrin-mediated adhesion to collagen. HMEC-1 expressing activated Notch4 do not have increased surface expression of beta 1-integrins. Rather, we demonstrate that Notch4-expressing cells display beta1-integrin in an active, high-affinity conformation. Furthermore, using function-activating beta 1 integrin antibodies, we demonstrate that activation of beta1-integrins is sufficient to inhibit VEGF-induced endothelial sprouting in vitro and angiogenesis in vivo. Our findings suggest that constitutive Notch4 activation in endothelial cells inhibits angiogenesis in part by promoting beta 1-integrin mediated adhesion to the underlying matrix. PMID- 11909977 TI - Translational control of cell fate: availability of phosphorylation sites on translational repressor 4E-BP1 governs its proapoptotic potency. AB - Translational control has been recently added to well-recognized genomic, transcriptional, and posttranslational mechanisms regulating apoptosis. We previously found that overexpressed eukaryotic initiation factor 4E (eIF4E) rescues cells from apoptosis, while ectopic expression of wild-type eIF4E-binding protein 1 (4E-BP1), the most abundant member of the 4E-BP family of eIF4E repressor proteins, activates apoptosis--but only in transformed cells. To test the possibility that nontransformed cells require less cap-dependent translation to suppress apoptosis than do their transformed counterparts, we intensified the level of translational repression in nontransformed fibroblasts. Here, we show that inhibition of 4E-BP1 phosphorylation by rapamycin triggers apoptosis in cells ectopically expressing wild-type 4E-BP1 and that expression of 4E-BP1 phosphorylation site mutants potently activates apoptosis in a phosphorylation site-specific manner. In general, proapoptotic potency paralleled repression of cap-dependent translation. However, this relationship was not a simple monotone. As repression of cap-dependent translation intensified, apoptosis increased to a maximum value. Further repression resulted in less apoptosis--a state associated with activation of translation through internal ribosomal entry sites. These findings show: that phosphorylation events govern the proapoptotic potency of 4E BP1, that 4E-BP1 is proapoptotic in normal as well as transformed fibroblasts, and that malignant transformation is associated with a higher requirement for cap dependent translation to inhibit apoptosis. Our results suggest that 4E-BP1 mediated control of apoptosis occurs through qualitative rather than quantitative changes in protein synthesis, mediated by a dynamic interplay between cap dependent and cap-independent processes. PMID- 11909976 TI - Requirement of TRAP/mediator for both activator-independent and activator dependent transcription in conjunction with TFIID-associated TAF(II)s. AB - The multiprotein human TRAP/Mediator complex, which is phylogenetically related to the yeast SRB/Mediator coactivator, facilitates activation through a wide variety of transcriptional activators. However, it remains unclear how TRAP/Mediator functions in the context of other coactivators. Here we have identified a previously uncharacterized integral subunit (TRAP25) of the complex that is apparently metazoan specific. An antibody that is specific for TRAP25 allowed quantitative immunodepletion of essentially all TRAP/Mediator components from HeLa nuclear extract, without detectably affecting levels of RNA polymerase II and corresponding general transcription factors. Surprisingly, the TRAP/Mediator-depleted nuclear extract displayed severely reduced levels of both basal and activator-dependent transcription from DNA templates. Both activities were efficiently restored upon readdition of purified TRAP/Mediator. Moreover, restoration of basal and activator-dependent transcription to extracts that were simultaneously depleted of TRAP/Mediator and TFIID (TBP plus the major TAF(II)s) required addition of both TBP and associated TAF(II)s, as well as TRAP/Mediator. These observations indicate that TAF(II)s and Mediator are jointly required for both basal and activated transcription in the context of a more physiological complement of nuclear proteins. We propose a close mechanistic linkage between these components that most likely operates at the level of combined effects on the general transcription machinery and, in addition, a direct role for Mediator in relaying activation signals to this machinery. PMID- 11909980 TI - Pharmacology of dopamine agonists in the treatment of Parkinson's disease. AB - There is now increasing use of dopamine agonists as effective early monotherapy in the treatment of Parkinson's disease (PD). Dopamine agonists can induce an antiparkinsonian effect through actions on either D(1)-like or D(2)-like dopamine receptors, and the multiple receptor subtypes present in the brain may provide further opportunities to improve the treatment of PD. Functional interactions exist between D(1)- and D(2)-like receptors, and adaptive changes occur after denervation and repeated administration of a dopamine agonist. Long-acting dopamine agonists produce a lower incidence of dyskinesia than levodopa (L-dopa) when they are used as monotherapy in either PD or in drug-naive 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP)-treated primates. Continuous dopaminergic stimulation appears less likely to prime basal ganglia for involuntary movements compared with drugs, such as L-dopa, that produce pulsatile stimulation. However, once priming has occurred, dopamine agonists produce dyskinesia identical to that of L-dopa. Continuous administration of long-acting dopamine agonists may reverse the priming process initiated by L-dopa, markedly decreasing dyskinesia intensity with a minimal loss of antiparkinsonian activity, at least in MPTP-treated primates. Dopamine receptors in brain areas other than the striatum, such as the globus pallidus and subthalamic nucleus, and in the mesolimbic and mesocortical regions may also contribute to antiparkinsonian activity of dopamine agonists and their associated side effects. The future potential of dopamine agonists may lie in the selective stimulation of dopamine receptor subtypes in different brain areas and through the actions of partial dopamine agonists and drugs that normalize dopamine receptor function. PMID- 11909978 TI - NF-kappa B activates prostate-specific antigen expression and is upregulated in androgen-independent prostate cancer. AB - The transcription factor NF-kappa B regulates gene expression involved in cell growth and survival and has been implicated in progression of hormone-independent breast cancer. By expressing a dominant-active form of mitogen-activated protein kinase kinase kinase 1, by exposure to tumor necrosis factor alpha, or by overexpression of p50/p65, we show that NF-kappa B activates a transcription regulatory element of the prostate-specific antigen (PSA)-encoding gene, a marker for prostate cancer development, treatment, and progression. By DNase I footprinting, we identified four NF-kappa B binding sites in the PSA core enhancer. We also demonstrate that androgen-independent prostate cancer xenografts have higher constitutive NF-kappa B binding activity than their androgen-dependent counterparts. These results suggest a role of NF-kappa B in prostate cancer progression. PMID- 11909979 TI - MSK1 and MSK2 are required for the mitogen- and stress-induced phosphorylation of CREB and ATF1 in fibroblasts. AB - Using mouse knockouts for mitogen- and stress-activated protein kinase 1 (MSK1) and MSK2 and a double knockout of both MSK1 and MSK2, we show that these protein kinases are required for the stress-induced phosphorylation of transcription factors CREB and ATF1 in primary embryonic fibroblasts. In contrast mitogen induced phosphorylation of CREB and ATF1 is greatly reduced but not totally abolished. The mitogen- and stress-induced phosphorylation of CREB at Ser133 has been linked to the transcription of several immediate early genes, including c fos, junB, and egr1. The knockout of both MSK1 and MSK2 resulted in a 50% reduction in c-fos and junB gene transcription in response to anisomycin or UV-C radiation but only a small reduction in response to tetradecanoyl phorbol acetate or epidermal growth factor in fibroblasts. The transcription of egr1 in response to both mitogenic and stress stimuli, as well as stress-induced apoptosis, was unaffected in the MSK1/MSK2 double knockout. PMID- 11909982 TI - Levodopa strengths and weaknesses. AB - Despite clinical experience with levodopa for more than three decades, the role of this agent in the treatment of Parkinson's disease (PD) has not been well defined. Clearly the most effective antiparkinsonian drug, levodopa, is associated with emergence of motor complications, particularly fluctuations and dyskinesias, as well as other side effects. In addition to these limitations, there is an ongoing debate about the potential neurotoxic effects of levodopa, suggested by some in vitro studies. However, there is no support for levodopa induced neurotoxicity from in vivo studies. This review discusses possible mechanisms of levodopa-related complications and therapeutic strategies for their prevention and management. PMID- 11909981 TI - Neuroprotection and dopamine agonists. AB - Several factors are known to be capable of inducing relatively selective dopaminergic cell death in the substantia nigra and inducing the clinical features that characterize Parkinson's disease (PD). Neuronal toxins such as 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) can induce parkinsonism in human and animal models, and rotenone, another specific mitochondrial complex I inhibitor, can induce similar effects in rodents to produce a model for PD. Studies in twins suggest a significant genetic component to young-onset PD, and several gene mutations have now been identified as causing familial autosomal dominant or autosomal recessive PD. Etiologic factors including free radical mediated damage (including excitotoxicity), mitochondrial dysfunction, and inflammation-mediated cell damage can contribute to pathogenesis. In addition, the recent interest in protein misfolding, aggregation, and proteosomal activity has provided further insight into potential pathogenetic pathways in PD. Against this background there has been increasing interest in the development of drugs to modify these biochemical abnormalities and thus alter the course of PD, either by retarding the rate of cell death or by restoring function to neurons that are likely to be damaged but not dead. In this context, dopamine agonists have shown significant promise. Not only do these drugs provide symptomatic relief of PD but they also appear to be associated with a significant decrease in the rate of motor complications and to be capable of protecting against some of the adverse consequences of levodopa use. However, evidence is now emerging that dopamine agonists may have additional neuroprotective properties. As a group, they have antioxidant actions in vitro and in vivo. More specifically, the D(2)/D(3) dopamine agonist pramipexole may have neuroprotective activity that is, at least in part, unrelated to its dopamine agonist action. Protection in cell and animal models against a variety of toxins, including MPTP and 6-hydroxydopamine, confirms that this agonist has in vitro and in vivo neuroprotective action. Evidence is now emerging that some of this may be mediated by direct action on mitochondrial membrane potential and the inhibition of apoptosis. If the neuroprotective action of this drug is confirmed in patients with PD, this will have important implications for its early use in patients. PMID- 11909984 TI - Long-term studies of dopamine agonists. AB - Dopamine agonists have long been used as adjunctive therapy for the treatment of Parkinson's disease (PD). In more recent years these drugs have also been proved safe and effective as initial therapy in lieu of levodopa in the treatment of PD. Long-term levodopa therapy is associated with motor complications, including fluctuating response patterns and dyskinesia. By initially introducing a dopamine agonist as symptomatic drug therapy, it may be possible to postpone the use of levodopa and delay or prevent the development of motor complications. Recently, four clinical trials have explored this hypothesis by comparing the long-term response and side effects of levodopa with dopamine agonist therapy. The drugs studied have included ropinirole, pramipexole, cabergoline, and pergolide. In each of these projects, the occurrence of motor complications, such as wearing off and dyskinesia, was significantly less in the subjects assigned to initiation of therapy with a dopamine agonist. The addition of levodopa could be postponed by many months or even several years. Therefore, these long-term studies of dopamine agonists support the initiation of a dopamine agonist instead of levodopa in an effort to postpone levodopa-related motor complications. This therapeutic approach may be particularly appropriate in PD patients with a long treatment horizon on the basis of age and general good health. The extension phase of the long-term study comparing pramipexole with levodopa is ongoing, and follow-up information may help to establish the value of this treatment strategy. PMID- 11909983 TI - The role of dopamine agonists in the treatment of early Parkinson's disease. AB - Levodopa is the gold standard for the symptomatic treatment of Parkinson's disease (PD). However, chronic treatment is associated with development of motor complications in the majority of patients. Recent laboratory studies suggest that pulsatile administration of a short-acting agent, such as levodopa, contributes to the development of these problems and that they might be mitigated through the use of longer-acting dopamine agonists. Placebo-controlled clinical trials have shown that dopamine agonists have anti-parkinsonian effects in patients with early PD. More importantly, four different prospective double-blind studies have demonstrated that initiating symptomatic therapy with a dopamine agonist is associated with a significantly reduced risk for development of motor complications than is initial treatment with a dopamine agonist. Furthermore, several lines of laboratory research suggest that dopamine agonists might protect dopaminergic neurons in PD and retard the rate of disease progression. Double blind trials using clinical and imaging end points are now testing this hypothesis and the results should soon be available. On the basis of this evidence, a rational approach to the treatment of PD patients who are not elderly or cognitively impaired is to initiate therapy with a dopamine agonist and supplement with levodopa when dopamine agonist monotherapy can no longer provide satisfactory clinical control. PMID- 11909985 TI - Relevance of motor complications in Parkinson's disease. AB - Although patients with Parkinson's disease usually respond to dopaminergic therapy with a smooth, continued effect when medication is first initiated, many patients eventually develop a fluctuating response along with involuntary movements (dyskinesias). The fluctuations in motor response often result in patients requiring more frequent dosing of medication that is less convenient, and they begin to lose control of their daily life. Their ability to work or to perform activities of daily living may fluctuate with response to medication, and the involuntary movements may interfere with activities. The fluctuations and dyskinesias can be painful and quite embarrassing. Overall, quality of life suffers. In addition, there is a marked increase in the expense to both the patient and society when motor fluctuations and dyskinesias develop. PMID- 11909986 TI - Tremor and dopamine agonists. AB - Although all dopaminergic drugs are effective in reducing tremor, no single drug has been shown to be clearly superior in the treatment of tremor. Levodopa produces a mean improvement of 30 to 50% in the Unified Parkinson's Disease Rating Scale (UPDRS) subtest for rest tremor. Comparable improvement is achieved with the dopamine agonists. Dopamine agonists are particularly well suited for patients with newly diagnosed tremor-predominant disease and no cognitive impairment, but they are also useful in advanced patients with tremor that is refractory to levodopa and anticholinergics. The response of tremor to pharmacotherapy is variable, and clinicians must be prepared to try all of the available drugs before concluding that surgery is the only alternative. PMID- 11909987 TI - Depression associated with Parkinson's disease: clinical features and treatment. AB - Depression associated with Parkinson's disease (PD) is common and affects 25 to 40% of patients. Recognition of the signs and symptoms of depression associated with PD is essential for clinical practitioners. Treatment of depression in this subset of patients can have a direct and dramatic impact on functional disability and quality of life. A review of the literature concerning depression and depression associated with PD was undertaken, with specific attention given to disease mechanisms, clinical presentation, association with thyroid disease, and the principles of management and treatment. Specific signs and symptoms of depression can be easily identified in patients with PD. Practitioners should be aware of these signs and symptoms when diagnosing and treating depression associated with PD. Practitioners should also be aware of the pros and cons of each treatment option and should choose a therapy appropriate for each individual patient's needs. It is important to identify the features of depression associated with PD in order to render early diagnosis and institute practical and efficacious therapy. PMID- 11909988 TI - Dopamine agonists and sleep in Parkinson's disease. AB - Dopaminergic therapy is increasingly recognized as a cause of excessive daytime sleepiness in patients with PD. This adverse effect may be a dose-related phenomenon that is somewhat more likely to occur with dopamine agonists than with levodopa, although all dopaminergic drugs can be sedating. However, medication effect is only one of several causes of somnolence in PD. Other factors include age-related changes in sleep quality, nocturnal motor disturbances, primary sleep disorders such as sleep apnea, medication-induced sleep disruption, and concurrent medical illnesses. There is also increasing evidence that the disease process itself may affect the control of the sleep-wake cycle. Although we have characterized the sleep disturbances in PD, further investigation is needed to define their prevalence and etiology, particularly with respect to the role of dopamine and dopaminergic agents. Clinicians should be alert to the complaint of excessive sleepiness in their patients and should attempt to identify and treat the underlying causes. PMID- 11909989 TI - Treatment of early Parkinson's disease. AB - The early treatment of Parkinson's disease (PD) consists of nonpharmacologic treatment, consideration of neuroprotective therapy, and initial symptomatic treatment. Education for the patient and family, access to support groups, regular exercise, and good nutrition are essential to the overall management of PD. Disease-modifying therapies, such as agents that provide neurorescue or neuroprotection, will provide a major advance in the treatment of PD. Intervention at the genetic/environmental level or that affects the cascade of pathophysiologic events, protein aggregation, or apoptosis could result in neuroprotection. Many agents are now being investigated for neuroprotective potential. A major paradigm shift has recently occurred because of the recent basic and clinical data indicating that dopamine agonists, rather than levodopa, should be the initial symptomatic therapy in PD. However, levodopa may be started first in some patients because of patient age, cognitive status, or cost of drugs. PMID- 11909991 TI - Why doesn't the elephant have a pleural space? AB - The elephant is the only mammal whose pleural space is obliterated by connective tissue. This has been known for 300 years but never explained. The elephant is also the only animal that can snorkel at depth. The resulting pressure differences require changes in the pleural membranes and pleural space. PMID- 11909990 TI - Restless legs syndrome: treatment with dopaminergic agents. AB - Restless legs syndrome (RLS) is a common neurologic disorder that affects 5 to 10% of the population and increases in prevalence with aging. The clinical hallmarks of RLS include dysesthesias or paresthesias in the legs and sometimes the arms, occurring primarily at rest, which are usually worse in the evening and are alleviated by movement. RLS can be a disabling disorder, causing sleep disturbance at night and excessive sleepiness during the day. Although treatment with levodopa alleviates symptoms, many RLS patients develop rebound (occurrence of symptoms during the night) or augmentation (occurrence of symptoms before levodopa dosing in the evening). Augmentation occurs in up to 82% of patients treated with levodopa, limiting the long-term usefulness of this agent. The direct dopamine receptor agonists are long-acting drugs often administered as a single dose at bedtime. Among these agents, pergolide, pramipexole, ropinirole, and cabergoline have all been shown to alleviate RLS symptoms in 70 to 100% of patients. The most common adverse effect is nausea. Augmentation, although it may be associated with chronic agonist use, is usually mild and responsive to additional dosing. The direct dopamine receptor agonists have largely replaced levodopa as the most effective treatment for RLS. PMID- 11909992 TI - The basal ganglia and disorders of movement: pathophysiological mechanisms. AB - The basal ganglia are part of a neuronal network organized in parallel circuits. The "motor circuit" is most relevant to the pathophysiology of movement. Abnormal increment or reduction in the inhibitory output activity of basal ganglia give rise, respectively, to poverty and slowness of movement (i.e., Parkinson's disease) or dyskinesias. PMID- 11909993 TI - Plasminogen activator inhibitor 1: physiological and pathophysiological roles. AB - Plasminogen activator inhibitor 1 (PAI-1) inhibits plasminogen activators (u-PA and t-PA) by forming stable complexes endocytosed via a low-density lipoprotein receptor superfamily member-dependent mechanism. PAI-1 circulates actively in plasma and latently in platelets but is also secreted and deposited into the matrix by several cells, where it participates in tissue repair processes. PMID- 11909994 TI - MAP kinases: from intracellular signals to physiology and disease. AB - Although differentiated cells will usually maintain their specialized character, conversion of cellular specificities can be observed during adaptation or reparative regeneration. In pathological conditions, such as inflammation and carcinogenesis, even highly specialized cells can alter their properties, leading to a deranged control of cell differentiation and/or proliferation. Mitogen activated protein kinases are central regulators of these processes. PMID- 11909995 TI - New roles for old holes: ion channel function in aquaporin-1. AB - Mammalian aquaporins are part of the diverse major intrinsic protein family of water and solute channels. Intriguing links exist in structural and functional properties between aquaporins and ion channels. A novel role for aquaporin-1 as a gated ion channel reshapes our current views of this ancient family of transmembrane channel proteins. PMID- 11909996 TI - ADAM-mediated shedding and adhesion: a vascular perspective. AB - Proteolytic conversion (shedding) of membrane-bound proteins to soluble forms is a novel regulatory mechanism mediated by enzymes called a disintegrin and metalloproteases (ADAMs). In this review, the potential importance of ADAMs in vascular physiology and disease, especially in relation to atherosclerosis, will be highlighted. PMID- 11909997 TI - Arterial gas embolism and decompression sickness. AB - Decompression sickness occurs when a sufficiently large gas phase forms within the tissues of the body after a reduction in ambient pressure. Arterial gas embolism occurs secondary to pulmonary barotrauma when gas is forced into the pulmonary vasculature. Although they may clinically present in a similar fashion, the underlying pathophysiology of the two conditions is quite different. PMID- 11909998 TI - Insulin-like growth factor-I: compartmentalization within the somatotropic axis? AB - Insulin-like growth factor-I (IGF-I) is essential for normal growth; igf-1 gene mutations are associated with extreme growth retardation in mice and, very rarely, in humans. The relative contributions of tissue vs. endocrine (hepatic) IGF-I to the regulation of growth has been a fundamental question. New gene targeting technologies are providing answers for these questions. PMID- 11910000 TI - Mapping and sequencing the rice genome. PMID- 11910001 TI - A comprehensive rice transcript map containing 6591 expressed sequence tag sites. AB - To determine the chromosomal positions of expressed rice genes, we have performed an expressed sequence tag (EST) mapping project by polymerase chain reaction based yeast artificial chromosome (YAC) screening. Specific primers designed from 6713 unique EST sequences derived from 19 cDNA libraries were screened on 4387 YAC clones and used for map construction in combination with genetic analysis. Here, we describe the establishment of a comprehensive YAC-based rice transcript map that contains 6591 EST sites and covers 80.8% of the rice genome. Chromosomes 1, 2, and 3 have relatively high EST densities, approximately twice those of chromosomes 11 and 12, and contain 41% of the total EST sites on the map. Most of the EST-dense regions are distributed on the distal regions of each chromosome arm. Genomic regions flanking the centromeres for most of the chromosomes have lower EST density. Recombination frequency in these regions is suppressed significantly. Our EST mapping also shows that 40% of the assigned ESTs occupy only approximately 21% of the entire genome. The rice transcript map has been a valuable resource for genetic study, gene isolation, and genome sequencing at the Rice Genome Research Program and should become an important tool for comparative analysis of chromosome structure and evolution among the cereals. PMID- 11910002 TI - An integrated physical and genetic map of the rice genome. AB - Rice was chosen as a model organism for genome sequencing because of its economic importance, small genome size, and syntenic relationship with other cereal species. We have constructed a bacterial artificial chromosome fingerprint-based physical map of the rice genome to facilitate the whole-genome sequencing of rice. Most of the rice genome ( approximately 90.6%) was anchored genetically by overgo hybridization, DNA gel blot hybridization, and in silico anchoring. Genome sequencing data also were integrated into the rice physical map. Comparison of the genetic and physical maps reveals that recombination is suppressed severely in centromeric regions as well as on the short arms of chromosomes 4 and 10. This integrated high-resolution physical map of the rice genome will greatly facilitate whole-genome sequencing by helping to identify a minimum tiling path of clones to sequence. Furthermore, the physical map will aid map-based cloning of agronomically important genes and will provide an important tool for the comparative analysis of grass genomes. PMID- 11910003 TI - KNAT1 and ERECTA regulate inflorescence architecture in Arabidopsis. AB - Plant architecture is dictated by morphogenetic factors that specify the number and symmetry of lateral organs as well as their positions relative to the primary axis. Mutants defective in the patterning of leaves and floral organs have provided new insights on the signaling pathways involved, but there is comparatively little information regarding aspects of the patterning of stems, which play a dominant role in architecture. To this end, we have characterized five alleles of the brevipedicellus mutant of Arabidopsis, which exhibits reduced internode and pedicel lengths, bends at nodes, and downward-oriented flowers and siliques. Bends in stems correlate with a loss of chlorenchyma tissue at the node adjacent to lateral organs and in the abaxial regions of pedicels. A stripe of achlorophyllous tissue extends basipetally from each node and is positioned over the vasculature that services the corresponding lateral organ. Map-based cloning and complementation studies revealed that a null mutation in the KNAT1 homeobox gene is responsible for these pleiotropic phenotypes. Our observation that wild type Arabidopsis plants also downregulate chlorenchyma development adjacent to lateral organs leads us to propose that KNAT1 and ERECTA are required to restrict the action of an asymmetrically localized, vasculature-associated chlorenchyma repressor at the nodes. Our data indicate that it is feasible to alter the architecture of ornamental and crop plants by manipulating these genetically defined pathways. PMID- 11910004 TI - Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. AB - Numerous studies have shown that transcription factors are important in regulating plant responses to environmental stress. However, specific functions for most of the genes encoding transcription factors are unclear. In this study, we used mRNA profiles generated from microarray experiments to deduce the functions of genes encoding known and putative Arabidopsis transcription factors. The mRNA levels of 402 distinct transcription factor genes were examined at different developmental stages and under various stress conditions. Transcription factors potentially controlling downstream gene expression in stress signal transduction pathways were identified by observed activation and repression of the genes after certain stress treatments. The mRNA levels of a number of previously characterized transcription factor genes were changed significantly in connection with other regulatory pathways, suggesting their multifunctional nature. The expression of 74 transcription factor genes responsive to bacterial pathogen infection was reduced or abolished in mutants that have defects in salicylic acid, jasmonic acid, or ethylene signaling. This observation indicates that the regulation of these genes is mediated at least partly by these plant hormones and suggests that the transcription factor genes are involved in the regulation of additional downstream responses mediated by these hormones. Among the 43 transcription factor genes that are induced during senescence, 28 of them also are induced by stress treatment, suggesting extensive overlap responses to these stresses. Statistical analysis of the promoter regions of the genes responsive to cold stress indicated unambiguous enrichment of known conserved transcription factor binding sites for the responses. A highly conserved novel promoter motif was identified in genes responding to a broad set of pathogen infection treatments. This observation strongly suggests that the corresponding transcription factors play general and crucial roles in the coordinated regulation of these specific regulons. Although further validation is needed, these correlative results provide a vast amount of information that can guide hypothesis-driven research to elucidate the molecular mechanisms involved in transcriptional regulation and signaling networks in plants. PMID- 11910006 TI - AUX1 promotes lateral root formation by facilitating indole-3-acetic acid distribution between sink and source tissues in the Arabidopsis seedling. AB - Arabidopsis root architecture is regulated by shoot-derived signals such as nitrate and auxin. We report that mutations in the putative auxin influx carrier AUX1 modify root architecture as a result of the disruption in hormone transport between indole-3-acetic acid (IAA) source and sink tissues. Gas chromatography selected reaction monitoring-mass spectrometry measurements revealed that the aux1 mutant exhibited altered IAA distribution in young leaf and root tissues, the major IAA source and sink organs, respectively, in the developing seedling. Expression studies using the auxin-inducible reporter IAA2::uidA revealed that AUX1 facilitates IAA loading into the leaf vascular transport system. AUX1 also facilitates IAA unloading in the primary root apex and developing lateral root primordium. Exogenous application of the synthetic auxin 1-naphthylacetic acid is able to rescue the aux1 lateral root phenotype, implying that root auxin levels are suboptimal for lateral root primordium initiation in the mutant. PMID- 11910005 TI - The Arabidopsis salt overly sensitive 4 mutants uncover a critical role for vitamin B6 in plant salt tolerance. AB - Salt stress is a major environmental factor influencing plant growth and development. To identify salt tolerance determinants, a genetic screen for salt overly sensitive (sos) mutants was performed in Arabidopsis. We present here the characterization of sos4 mutants and the positional cloning of the SOS4 gene. sos4 mutant plants are hypersensitive to Na(+), K(+), and Li(+) ions. Under NaCl stress, sos4 plants accumulate more Na(+) and retain less K(+) compared with wild type plants. SOS4 encodes a pyridoxal kinase that is involved in the biosynthesis of pyridoxal-5-phosphate, an active form of vitamin B6. The expression of SOS4 cDNAs complements an Escherichia coli mutant defective in pyridoxal kinase. Supplementation of pyridoxine but not pyridoxal in the growth medium can partially rescue the sos4 defect in salt tolerance. SOS4 is expressed ubiquitously in all plant tissues. As a result of alternative splicing, two transcripts are derived from the SOS4 gene, the relative abundance of which is modulated by development and environmental stresses. Besides being essential cofactors for numerous enzymes, as shown by pharmacological studies in animal cells, pyridoxal-5-phosphate and its derivatives are also ligands for P2X receptor ion channels. Our results demonstrate that pyridoxal kinase is a novel salt tolerance determinant important for the regulation of Na(+) and K(+) homeostasis in plants. We propose that pyridoxal-5-phosphate regulates Na(+) and K(+) homeostasis by modulating the activities of ion transporters. PMID- 11910008 TI - Cell pattern in the Arabidopsis root epidermis determined by lateral inhibition with feedback. AB - In the root epidermis of Arabidopsis, hair and nonhair cell types are specified in a distinct position-dependent pattern. Here, we show that transcriptional feedback loops between the WEREWOLF (WER), CAPRICE (CPC), and GLABRA2 (GL2) genes help to establish this pattern. Positional cues bias the expression of the WER MYB gene, leading to the induction of CPC and GL2 in cells located in a particular position (N) and adoption of the nonhair fate. The truncated MYB encoded by CPC mediates a lateral inhibition mechanism to negatively regulate WER, GL2, and its own gene in the alternative position (H) to induce the hair fate. These results provide a molecular genetic framework for understanding the determination of a cell-type pattern in plants. PMID- 11910007 TI - Establishment of cereal endosperm expression domains: identification and properties of a maize transfer cell-specific transcription factor, ZmMRP-1. AB - In maize, cells at the base of the endosperm are transformed into transfer cells that facilitate nutrient uptake by the developing seed. ZmMRP-1 is the first transfer cell-specific transcriptional activator to be identified. The protein it encodes contains nuclear localization signals and a MYB-related DNA binding domain. A single gene copy is present in maize, mapping to a locus on chromosome 8. ZmMRP-1 is first expressed soon after fertilization, when the endosperm is still a multinuclear coenocyte. The transcript accumulates in the basal nucleocytoplasmic domain that gives rise to transfer cells after cellularization. The transcript can be detected throughout transfer cell development, but it is not found in mature cells. ZmMRP-1 strongly transactivates the promoters of two unrelated transfer cell-specific genes. The properties of ZmMRP-1 are consistent with it being a determinant of transfer cell-specific expression. Possible roles for ZmMRP-1 in the regulation of endosperm and transfer cell differentiation are discussed. PMID- 11910009 TI - Rice SPK, a calmodulin-like domain protein kinase, is required for storage product accumulation during seed development: phosphorylation of sucrose synthase is a possible factor. AB - Suc, an end product of photosynthesis, is metabolized by Suc synthase in sink organs as an initial step in the biosynthesis of storage products. Suc synthase activity is known to be regulated by reversible phosphorylation, but the details of this process are unclear at present. Rice SPK, a calcium-dependent protein kinase, is expressed uniquely in the endosperm of immature seed, and its involvement in the biosynthetic pathways of storage products was suggested. Antisense SPK transformants lacked the ability to accumulate storage products such as starch, but produced watery seed with a large amount of Suc instead, as the result of an inhibition of Suc degradation. Analysis of in vitro phosphorylation indicated that SPK phosphorylated specifically a Ser residue in Suc synthase that has been shown to be important for its activity in the degradation of Suc. This finding suggests that SPK is involved in the activation of Suc synthase. It appears that SPK is a Suc synthase kinase that may be important for supplying substrates for the biosynthesis of storage products. PMID- 11910010 TI - Fertile hypomorphic ARGONAUTE (ago1) mutants impaired in post-transcriptional gene silencing and virus resistance. AB - Transgene-induced post-transcriptional gene silencing (PTGS) results from specific degradation of RNAs that are homologous with the transgene transcribed sequence. This phenomenon, also known as cosuppression in plants and quelling in fungi, resembles RNA interference (RNAi) in animals. Indeed, cosuppression/quelling/RNAi require related PAZ/PIWI proteins (AGO1/QDE-2/RDE-1), indicating that these mechanisms are related. Unlike Neurospora crassa qde-2 and Caenorhabditis elegans rde-1 mutants, which are morphologically normal, the 24 known Arabidopsis ago1 mutants display severe developmental abnormalities and are sterile. Here, we report the isolation of hypomorphic ago1 mutants, including fertile ones. We show that these hypomorphic ago1 mutants are defective for PTGS, like null sgs2, sgs3, and ago1 mutants, suggesting that PTGS is more sensitive than development to perturbations in AGO1. Conversely, a mutation in ZWILLE/PINHEAD, another member of the Arabidopsis AGO1 gene family, affects development but not PTGS. Similarly, mutations in ALG-1 and ALG-2, two members of the C. elegans RDE-1 gene family, affect development but not RNAi, indicating that the control of PTGS/RNAi and development by PAZ/PIWI proteins can be uncoupled. Finally, we show that hypomorphic ago1 mutants are hypersensitive to virus infection, confirming the hypothesis that in plants PTGS is a mechanism of defense against viruses. PMID- 11910011 TI - In vivo analysis of the role of atTic20 in protein import into chloroplasts. AB - The import of nucleus-encoded preproteins into plastids requires the coordinated activities of membrane protein complexes that facilitate the translocation of polypeptides across the envelope double membrane. Tic20 was identified previously as a component of the import machinery of the inner envelope membrane by covalent cross-linking studies with trapped preprotein import intermediates. To investigate the role of Tic20 in preprotein import, we altered the expression of the Arabidopsis Tic20 ortholog (atTic20) by antisense expression. Several antisense lines exhibited pronounced chloroplast defects exemplified by pale leaves, reduced accumulation of plastid proteins, and significant growth defects. The severity of the phenotypes correlated directly with the reduction in levels of atTic20 expression. In vitro import studies with plastids isolated from control and antisense plants indicated that the antisense plastids are defective specifically in protein translocation across the inner envelope membrane. These data suggest that Tic20 functions as a component of the protein-conducting channel at the inner envelope membrane. PMID- 11910012 TI - Zein protein interactions, rather than the asymmetric distribution of zein mRNAs on endoplasmic reticulum membranes, influence protein body formation in maize endosperm. AB - Prolamin-containing protein bodies in maize endosperm are composed of four different polypeptides, the alpha-, beta-, gamma-, and delta-zeins. The spatial organization of zeins within the protein body, as well as interactions between them, suggests that the localized synthesis of gamma-zeins could initiate and target protein body formation at specific regions of the rough endoplasmic reticulum. To investigate this possibility, we analyzed the distribution of mRNAs encoding the 22-kD alpha-zein and the 27-kD gamma-zein proteins on cisternal and protein body rough endoplasmic reticulum membranes. In situ hybridization revealed similar frequencies of the mRNAs in both regions of the endoplasmic reticulum, indicating that the transcripts are distributed more or less randomly. This finding implies that zein protein interactions determine protein body assembly. To address this question, we expressed cDNAs encoding alpha-, beta-, gamma-, and delta-zeins in the yeast two-hybrid system. We found strong interactions among the 50-, 27-, and 16-kD gamma-zeins and the 15-kD beta-zein, consistent with their colocalization in developing protein bodies. Interactions between the 19- and 22-kD alpha-zeins were relatively weak, although each of them interacted strongly with the 10-kD delta-zein. Strong interactions were detected between the alpha- and delta-zeins and the 16-kD gamma-zein and the 15-kD beta zein; however, the 50- and 27-kD gamma-zeins did not interact with the alpha- and delta-zein proteins. We identified domains within the 22-kD alpha-zein that bound preferentially the alpha- and delta-zeins and the beta- and gamma-zeins. Affinities between zeins generally were consistent with results from immunolocalization experiments, suggesting an important role for the 16-kD gamma zein and the 15-kD beta-zein in the binding and assembly of alpha-zeins within the protein body. PMID- 11910013 TI - Reciprocal expression of two candidate di-iron enzymes affecting photosystem I and light-harvesting complex accumulation. AB - Crd1 (Copper response defect 1), which is required for the maintenance of photosystem I and its associated light-harvesting complexes in copper-deficient ( Cu) and oxygen-deficient (-O(2)) Chlamydomonas reinhardtii cells, is localized to the thylakoid membrane. A related protein, Cth1 (Copper target homolog 1), is shown to have a similar but not identical function by genetic suppressor analysis of gain-of-function sct1 (suppressor of copper target 1) strains that are transposon-containing alleles at CTH1. The pattern of Crd1 versus Cth1 accumulation is reciprocal; Crd1 abundance is increased in -Cu or -O(2) cells, whereas Cth1 accumulates in copper-sufficient (+Cu), oxygenated cells. This expression pattern is determined by a single trans-acting regulatory locus, CRR1 (COPPER RESPONSE REGULATOR 1), which activates transcription in -Cu cells. In +Cu cells, a 2.1-kb Cth1 mRNA is produced and translated, whereas Crd1 is transcribed only at basal levels, leading to Cth1 accumulation in +Cu cells. In -Cu cells, CRR1 function determines the activation of Crd1 expression and the production of an alternative 3.1-kb Cth1 mRNA that is extended at the 5' end relative to the 2.1-kb mRNA. Synthesis of the 3.1-kb mRNA, which encodes six small upstream open reading frames that possibly result in poor translation, blocks the downstream promoter through transcriptional occlusion. Fluorescence analysis of wild-type, crd1, and sct1 strains indicates that copper-responsive adjustment of the Cth1:Crd1 ratio results in modification of the interactions between photosystem I and associated light-harvesting complexes. The tightly coordinated CRR1-dependent regulation of isoenzymes Cth1 and Crd1 reinforces the notion that copper plays a specific role in the maintenance of chlorophyll proteins. PMID- 11910014 TI - The Cf-9 disease resistance protein is present in an approximately 420-kilodalton heteromultimeric membrane-associated complex at one molecule per complex. AB - The tomato Cf-9 gene confers race-specific resistance to the fungal pathogen Cladosporium fulvum expressing the corresponding avirulence gene Avr9. In tobacco, Cf-9 confers a hypersensitive response to the Avr9 peptide. To investigate Cf-9 protein function in initiating defense signaling, we engineered a functional C-terminal fusion of the Cf-9 gene with the TAP (Tandem Affinity Purification) tag. In addition, we established a transient expression assay in Nicotiana benthamiana leaves for the production of functional Cf-9:myc and Cf 9:TAP. Transiently expressed Cf-9:myc and Cf-9:TAP proteins induced an Avr9 dependent hypersensitive response, consistent with previous results with stably transformed tobacco plants and derived cell suspension cultures expressing c-myc tagged Cf-9. Gel filtration of microsomal fractions solubilized with octylglucoside revealed that the Cf-9 protein, either as c-myc or TAP fusions, migrated at a molecular mass of 350 to 475 kD. By using blue native gel electrophoresis, the molecular size was confirmed to be approximately 420 kD. Our results suggest that only one Cf-9 protein molecule is present in the Cf-9 complex and that Cf-9 is part of a membrane complex consisting of an additional glycoprotein partner(s). The high structural similarity between Cf proteins and Clavata2 (CLV2) of Arabidopsis, together with the similarity of molecular mass between Cf-9 and CLV complexes (420 and 450 kD, respectively), led us to investigate whether Cf-9 is integrated into membrane-associated protein complexes like those formed by CLV1 and CLV2. Unlike CLV2, the Cf-9 protein did not form disulfide-linked heterodimers, no ligand (Avr9)-dependent shift in the molecular mass of the Cf-9 complex was detected, and no Rho-GTPase-related proteins were found associated with Cf-9 under the conditions tested. Thus, Cf-9-dependent defense signaling and CLV2-dependent regulation of meristem development seem to be accomplished via distinct mechanisms, despite the structural similarity of their key components Cf-9 and CLV2. PMID- 11910015 TI - Convergence and divergence of stress-induced mitogen-activated protein kinase signaling pathways at the level of two distinct mitogen-activated protein kinase kinases. AB - Plants respond to biotic and abiotic stresses by inducing overlapping sets of mitogen-activated protein kinases (MAPKs) and response genes. To define the mechanisms of how different signals can activate a common signaling pathway, upstream activators of SIMK, a salt stress- and pathogen-induced alfalfa MAPK, were identified. Here, we compare the properties of SIMKK, a MAPK kinase (MAPKK) that mediates the activation of SIMK by salt stress, with those of PRKK, a distantly related novel MAPKK. Although both SIMKK and PRKK show strongest interaction with SIMK, SIMKK can activate SIMK without stimulation by upstream factors. In contrast, PRKK requires activation by an upstream activated MAPKK kinase. SIMKK mediates pathogen elicitor signaling and salt stress, but PRKK transmits only elicitor-induced MAPK activation. Of four tested MAPKs, PRKK activates three of them (SIMK, MMK3, and SAMK) upon elicitor treatment of cells. However, PRKK is unable to activate any MAPK upon salt stress. In contrast, SIMKK activates SIMK and MMK3 in response to elicitor, but it activates only SIMK upon salt stress. These data show that (1) MAPKKs function as convergence points for stress signals, (2) MAPKKs activate multiple MAPKs, and (3) signaling specificity is obtained not only through the inherent affinities of MAPKK-MAPK combinations but also through stress signal-dependent intracellular mechanisms. PMID- 11910016 TI - Use of the transposon Ac as a gene-searching engine in the maize genome. AB - We show here that, although genes constitute only a small percentage of the maize genome, it is possible to identify them phenotypically as Ac receptor sites. Simple and efficient Ac transposition assays based on the well-studied endosperm markers bz and wx were used to generate a collection of >1300 independent Ac transposants. The majority of transposed Ac elements are linked to either the bz or the wx donor loci on chromosome 9. A few of the insertions produce obvious visible phenotypes, but most of them do not, suggesting that these populations will be more useful for reverse genetics than for forward transposon mutagenesis. An inverse polymerase chain reaction method was adapted for the isolation of DNA adjacent to the transposed Ac elements (tac sites). Most Ac insertions were into unique DNA. By sequencing tac sites and comparing the sequences to existing databases, insertions were identified in a number of putative maize genes. The expression of most of these genes was confirmed by RNA gel blot analysis. We report here the isolation and characterization of the first 46 tac sites from the two insertion libraries. PMID- 11910017 TI - Plasma membrane aquaporins in the motor cells of Samanea saman: diurnal and circadian regulation. AB - Leaf-moving organs, remarkable for the rhythmic volume changes of their motor cells, served as a model system in which to study the regulation of membrane water fluxes. Two plasma membrane intrinsic protein homolog genes, SsAQP1 and SsAQP2, were cloned from these organs and characterized as aquaporins in Xenopus laevis oocytes. Osmotic water permeability (P(f)) was 10 times higher in SsAQP2 expressing oocytes than in SsAQP1-expressing oocytes. SsAQP1 was found to be glycerol permeable, and SsAQP2 was inhibited by 0.5 mM HgCl(2) and by 1 mM phloretin. The aquaporin mRNA levels differed in their spatial distribution in the leaf and were regulated diurnally in phase with leaflet movements. Additionally, SsAQP2 transcription was under circadian control. The P(f) of motor cell protoplasts was regulated diurnally as well: the morning and/or evening P(f) increases were inhibited by 50 microM HgCl(2), by 2 mM cycloheximide, and by 250 microM phloretin to the noon P(f) level. Our results link SsAQP2 to the physiological function of rhythmic cell volume changes. PMID- 11910020 TI - Histidine pK(a) shifts and changes of tautomeric states induced by the binding of gallium-protoporphyrin IX in the hemophore HasA(SM). AB - The HasA(SM) hemophore, secreted by Serratia marcescens, binds free or hemoprotein bound heme with high affinity and delivers it to a specific outer membrane receptor, HasR. In HasA(SM), heme is held by two loops and coordinated to iron by two residues, His 32 and Tyr 75. A third residue His 83 was shown recently to play a crucial role in heme ligation. To address the mechanistic issues of the heme capture and release processes, the histidine protonation states were studied in both apo- and holo-forms of HasA(SM) in solution. Holo HasA(SM) was formed with gallium-protoporphyrin IX (GaPPIX), giving rise to a diamagnetic protein. By use of heteronuclear correlation NMR spectroscopy, the imidazole side-chain (15)N and (1)H resonances of the six HasA(SM) histidines were assigned and their pKa values and predominant tautomeric states according to pH were determined. We show that protonation states of the heme pocket histidines can modulate the nucleophilic character of the two axial ligands and, consequently, control the heme binding. In particular, the essential role of the His 83 is emphasized according to its direct interaction with Tyr 75. PMID- 11910018 TI - Structural genomics: a pipeline for providing structures for the biologist. PMID- 11910019 TI - Natively unfolded proteins: a point where biology waits for physics. AB - The experimental material accumulated in the literature on the conformational behavior of intrinsically unstructured (natively unfolded) proteins was analyzed. Results of this analysis showed that these proteins do not possess uniform structural properties, as expected for members of a single thermodynamic entity. Rather, these proteins may be divided into two structurally different groups: intrinsic coils, and premolten globules. Proteins from the first group have hydrodynamic dimensions typical of random coils in poor solvent and do not possess any (or almost any) ordered secondary structure. Proteins from the second group are essentially more compact, exhibiting some amount of residual secondary structure, although they are still less dense than native or molten globule proteins. An important feature of the intrinsically unstructured proteins is that they undergo disorder-order transition during or prior to their biological function. In this respect, the Protein Quartet model, with function arising from four specific conformations (ordered forms, molten globules, premolten globules, and random coils) and transitions between any two of the states, is discussed. PMID- 11910021 TI - Amino acid intrinsic alpha-helical propensities III: positional dependence at several positions of C terminus. AB - In this study, we have analyzed experimentally the helical intrinsic propensities of non-charged and non-aromatic residues at different C-terminal positions (C1, C2, C3) of an Ala-based peptide. The effect was found to be complex, resulting in extra stabilization or destabilization, depending on guest amino acid and position under consideration. Polar (Ser, Thr, Cys, Asn, and Gln) amino acids and Gly were found to have significantly larger helical propensities at several C terminal positions compared with the alpha-helix center (-1.0 kcal/mole in some cases). Some of the nonpolar residues, especially beta-branched ones (Val and Ile) are significantly more favorable at position C3 (-0.3 to -0.4 kcal/mole), although having minor differences at other C-terminal positions compared with the alpha-helix center. Leu has moderate (-0.1 to -0.2 kcal/mole) stabilization effects at position C2 and C3, whereas being relatively neutral at C1. Finally, Met was found to be unfavorable at C1 and C2 ( +0.2 kcal/mole) and favorable at C3 (-0.2 kcal/mole). Thus, significant differences found between the intrinsic helical propensities at the C-terminal positions and those in the alpha-helix center must be accounted for in helix/coil transition theories and in protein design. PMID- 11910023 TI - Classification of G-protein coupled receptors by alignment-independent extraction of principal chemical properties of primary amino acid sequences. AB - We have developed an alignment-independent method for classification of G-protein coupled receptors (GPCRs) according to the principal chemical properties of their amino acid sequences. The method relies on a multivariate approach where the primary amino acid sequences are translated into vectors based on the principal physicochemical properties of the amino acids and transformation of the data into a uniform matrix by applying a modified autocross-covariance transform. The application of principal component analysis to a data set of 929 class A GPCRs showed a clear separation of the major classes of GPCRs. The application of partial least squares projection to latent structures created a highly valid model (cross-validated correlation coefficient, Q(2) = 0.895) that gave unambiguous classification of the GPCRs in the training set according to their ligand binding class. The model was further validated by external prediction of 535 novel GPCRs not included in the training set. Of the latter, only 14 sequences, confined in rapidly expanding GPCR classes, were mispredicted. Moreover, 90 orphan GPCRs out of 165 were tentatively identified to GPCR ligand binding class. The alignment-independent method could be used to assess the importance of the principal chemical properties of every single amino acid in the protein sequences for their contributions in explaining GPCR family membership. It was then revealed that all amino acids in the unaligned sequences contributed to the classifications, albeit to varying extent; the most important amino acids being those that could also be determined to be conserved by using traditional alignment-based methods. PMID- 11910022 TI - Fine-tuning function: correlation of hinge domain interactions with functional distinctions between LacI and PurR. AB - LacI and PurR are highly homologous proteins. Their functional units are homodimers, with an N-terminal DNA binding domain that comprises the helix-turn helix (HTH), N-linker, and hinge regions from both monomers. Hinge structural changes are known to occur upon DNA dissociation but are difficult to monitor experimentally. The initial steps of hinge unfolding were therefore examined using molecular dynamics simulations, utilizing a truncated, chimeric protein comprising the LacI HTH/N-linker and PurR hinge. A terminal Gly-Cys-Gly was added to allow "dimerization" through disulfide bond formation. Simulations indicate that differences in LacI and PurR hinge primary sequence affect the quaternary structure of the hinge x hinge' interface. However, these alternate hinge orientations would be sterically restricted by the core domain. These results prompted detailed comparison of recently available DNA-bound structures for LacI and truncated LacI(1-62) with the PurR structure. Examination revealed that different N-linker and hinge contacts to the core domain of the partner monomer (which binds effector molecule) affect the juxtapositions of the HTH, N-linker, and hinge regions in the DNA binding domain. In addition, the two full-length repressors exhibit significant differences in the interactions between the core and the C-linker connection to the DNA binding domain. Both linkers and the hinge have been implicated in the allosteric response of these repressors. Intriguingly, one functional difference between these two proteins is that they exhibit opposite allosteric response to effector. Simulations and observed structural distinctions are correlated with mutational analysis and sequence information from the LacI/GalR family to formulate a mechanism for fine-tuning individual repressor function. PMID- 11910024 TI - Analysis of serine proteinase-inhibitor interaction by alanine shaving. AB - We analyzed the energetic importance of residues surrounding the hot spot (the P(1) position) of bovine pancreatic trypsin inhibitor (BPTI) in interaction with two proteinases, trypsin and chymotrypsin, by a procedure called molecular shaving. One to eight residues of the structural epitope, composed of two extended and exposed loops, were mutated to alanine(s). Although truncation of the side chains of residues surrounding the P(1) position to methyl groups caused a decrease in Delta G(den) values up to 6.4 kcal mole(-1), it did not influence the overall conformation of the inhibitor. We found that the replacement of up to six residues with alanines was fully additive at the level of protein stability. To analyze the influence of the structural epitope on the association energy, we determined association constants for BPTI variants and both enzymes and applied the additivity analysis. Shaving of two binding loops led to a progressive drop in the association energy, more pronounced for trypsin (decrease up to 9.6 kcal mole(-1)) than chymotrypsin (decrease up to 3.5 kcal mole(-1)). In the case of extensively mutated variants interacting with chymotrypsin, the association energies agreed very well with the values calculated from single mutational effects. However, when P(1)-neighboring residues were shaved to alanine(s), their contribution to the association energy was not fully removed because of the presence of methyl groups and main chain-main chain intermolecular hydrogen bonds. Moreover, the hot spot had a different contribution to the complex stability in the fully shaved BPTI variant compared with the wild type, which was caused by perturbations of the P(1)-S(1) electrostatic interaction. PMID- 11910025 TI - The interdigitated beta-helix domain of the P22 tailspike protein acts as a molecular clamp in trimer stabilization. AB - The P22 tailspike adhesin is an elongated thermostable trimer resistant to protease digestion and to denaturation in sodium dodecyl sulfate. Monomeric, dimeric, and protrimeric folding and assembly intermediates lack this stability and are thermolabile. In the native trimer, three right-handed parallel beta helices (residues 143-540), pack side-by-side around the three-fold axis. After residue 540, these single chain beta-helices terminate and residues 541-567 of the three polypeptide chains wrap around each other to form a three-stranded interdigitated beta-helix. Three mutants located in this region -- G546D, R563Q, and A575T -- blocked formation of native tailspike trimers, and accumulated soluble forms of the mutant polypeptide chains within cells. The substitutions R563Q and A575T appeared to prevent stable association of partially folded monomers. G546D, in the interdigitated region of the chain, blocked tailspike folding at the transition from the partially-folded protrimer to the native trimer. The protrimer-like species accumulating in the G546D mutant melted out at 42 degrees C and was trypsin and SDS sensitive. The G546D defect was not corrected by introduction of global suppressor mutations, which correct kinetic defects in beta-helix folding. The simplest interpretation of these results is that the very high thermostability (T(m) = 88 degrees C), protease and detergent resistance of the native tailspike acquired in the protrimer-to-trimer transition, depends on the formation of the three-stranded interdigitated region. This interdigitated beta-helix appears to function as a molecular clamp insuring thermostable subunit association in the native trimer. PMID- 11910026 TI - Hydroxynonenal inactivates cathepsin B by forming Michael adducts with active site residues. AB - Oxidation of plasma low-density lipoprotein (oxLDL) generates the lipid peroxidation product 4-hydroxy-2 nonenal (HNE) and also reduces proteolytic degradation of oxLDL and other proteins internalized by mouse peritoneal macrophages in culture. This leads to accumulation of undegraded material in lysosomes and formation of ceroid, a component of foam cells in atherosclerotic lesions. To explore the possibility that HNE contributes directly to the inactivation of proteases, structure-function studies of the lysosomal protease cathepsin B have been pursued. We found that treatment of mouse macrophages with HNE reduces degradation of internalized maleyl bovine serine albumin and cathepsin B activity. Purified bovine cathepsin B treated briefly with 15 microM HNE lost approximately 76% of its protease activity and also developed immunoreactivity with antibodies to HNE adducts in Western blot analysis. After stabilization of the potential Michael adducts by sodium borohydride reduction, modified amino acids were localized within the bovine cathepsin B protein structure by mass spectrometric analysis of tryptic peptides. Michael adducts were identified by tandem mass spectrometry at cathepsin B active site residues Cys 29 (mature A chain) and His 150 (mature B chain). Thus, covalent interaction between HNE and critical active site residues inactivates cathepsin B. These results support the hypothesis that the accumulation of undegraded macromolecules in lysosomes after oxidative damage are caused in part by direct protease inactivation by adduct formation with lipid peroxidation products such as HNE. PMID- 11910027 TI - Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method. AB - We recently reported on a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based technique, termed SUPREX, that removes several important limitations associated with measuring the thermodynamic stability of proteins. In contrast to conventional spectroscopy based techniques for characterizing the equilibrium unfolding behavior of proteins, SUPREX is amenable to the thermodynamic analysis of both purified and unpurified proteins using mg to ng quantities of material. Here we report on the application of SUPREX to the analysis of multimeric protein systems. Included in this work are the SUPREX results we obtained in studies on six model multimeric proteins including the GCN4p1 dimer, the coil-V(a)L(d) trimer, the 4 oxalocrotonate tautomerase (4-OT) hexamer, the Trp repressor (TrpR) dimer, the Arc repressor (ArcR) dimer, and an ArcR mutant (the (DOA20)ArcR) dimer which contained two destabilizing mutations including an Asp to Ala mutation at position 20 and an amide to ester bond mutation between amino acid (aa) residues 19 and 20. As part of the work described here, we present a new method for the analysis of SUPREX data that is generally applicable to both monomeric and multimeric protein systems. Our results on the model proteins in this study indicate that this new method can be used to determine folding free energies for proteins with the accuracy and precision of conventional spectroscopy-based methods. PMID- 11910028 TI - Probability-based protein secondary structure identification using combined NMR chemical-shift data. AB - For a long time, NMR chemical shifts have been used to identify protein secondary structures. Currently, this is accomplished through comparing the observed (1)H(alpha), (13)C(alpha), (13)C(beta), or (13)C' chemical shifts with the random coil values. Here, we present a new protocol, which is based on the joint probability of each of the three secondary structural types (beta-strand, alpha helix, and random coil) derived from chemical-shift data, to identify the secondary structure. In combination with empirical smooth filters/functions, this protocol shows significant improvements in the accuracy and the confidence of identification. Updated chemical-shift statistics are reported, on the basis of which the reliability of using chemical shift to identify protein secondary structure is evaluated for each nucleus. The reliability varies greatly among the 20 amino acids, but, on average, is in the order of: (13)C(alpha)>(13)C'>(1)H(alpha)>(13)C(beta)>(15)N>(1)H(N) to distinguish an alpha helix from a random coil; and (1)H(alpha)>(13)C(beta) >(1)H(N) approximately (13)C(alpha) approximately (13)C' approximately (15)N for a beta-strand from a random coil. Amide (15)N and (1)H(N) chemical shifts, which are generally excluded from the application, in fact, were found to be helpful in distinguishing a beta-strand from a random coil. In addition, the chemical-shift statistical data are compared with those reported previously, and the results are discussed. A JAVA User Interface program has been developed to make the entire procedure fully automated and is available via http://ccsr3150-p3.stanford.edu. PMID- 11910029 TI - Mass spectrometric analysis of the kinetics of in vivo rhodopsin phosphorylation. AB - On stimulation, rhodopsin, the light-sensing protein in the rod cells of the retina, is phosphorylated at several sites on its C terminus as the first step in deactivation. We have developed a mass spectrometry-based method to quantify the kinetics of phosphorylation at each site in vivo. After exposing either a freshly dissected mouse retina or the eye of an anesthetized mouse to a flash of light, phosphorylation and dephosphorylation reactions are terminated by rapidly homogenizing the retina or enucleated eye in 8 M urea. The C-terminal peptide containing all known phosphorylation sites is cleaved from rhodopsin, partially purified by ultracentrifugation, and analyzed by liquid chromatography coupled with mass spectrometry (LCMS). The mass spectrometer responds linearly to the peptide from 10 fmole to 100 pmole. The relative sensitivity to peptides with zero to five phosphates was determined using purified phosphopeptide standards. High pressure liquid chromatography (HPLC) coupled with tandem mass spectrometry (LCMS/MS) was used to distinguish the three primary sites of phosphorylation, Ser 334, Ser 338, and Ser 343. Peptides monophosphorylated on Ser 334 were separable from those monophosphorylated on Ser 338 and Ser 343 by reversed-phase HPLC. Although peptides monophosphorylated at Ser 338 and Ser 343 normally coelute, the relative amounts of each species in the single peak could be determined by monitoring the ratio of specific daughter ions characteristic of each peptide. Doubly phosphorylated rhodopsin peptides with different sites of phosphorylation also were distinguished by LCMS/MS. The sensitivity of these methods was evaluated by using them to measure rhodopsin phosphorylation stimulated either by light flashes or by continuous illumination over a range of intensities. PMID- 11910030 TI - Complex behavior in solution of homodimeric SecA. AB - SecA, a homodimeric protein involved in protein export in Escherichia coli, exists in the cell both associated with the membrane translocation apparatus and free in the cytosol. SecA is a multifunctional protein involved in protein localization and regulation of its own expression. To carry out these functions, SecA interacts with a variety of proteins, phospholipids, nucleotides, and nucleic acid and shows two enzymic activities. It is an ATPase and a helicase. Its role during protein localization involves interaction with the precursor polypeptides to be exported, the cytosolic chaperone SecB, and the SecY subunit of the membrane-associated translocase, as well as with acidic phospholipids. At the membrane, SecA undergoes a cycle of binding and hydrolysis of ATP coupled to conformational changes that result in translocation of precursors through the cytoplasmic membrane. The helicase activity of SecA and its affinity for its mRNA are involved in regulation of its own expression. SecA has been reported to exist in at least two conformational states during its functional cycle. Here we have used analytical centrifugation, as well as column chromatography coupled with multi-angle light scatter, to show that in solution SecA undergoes at least two monomer-dimer equilibrium reactions that are sensitive to temperature and to concentration of salt. PMID- 11910031 TI - Improved affinity of engineered streptavidin for the Strep-tag II peptide is due to a fixed open conformation of the lid-like loop at the binding site. AB - The Strep-tag II is a nine-amino acid peptide that was developed as an affinity tool for the purification of corresponding fusion proteins on streptavidin columns. The peptide recognizes the same pocket of streptavidin where the natural ligand is normally bound so that biotin or its chemical derivatives can be used for competitive elution. We report here the crystal structures of the streptavidin mutants '1' and '2,' which had been engineered for 10-fold higher affinity towards the Strep-tag II. Both streptavidin mutants carry mutations at positions 44, 45, and 47, that is, in a flexible loop region close to the binding site. The crystal structures of the two apo-proteins and their complexes with the Strep-tag II peptide were refined at resolutions below 2 A. Both in the presence and absence of the peptide, the lid-like loop next to the ligand pocket- comprising residues 45 through 52--adopts an 'open' conformation in all four subunits within the asymmetric unit. The same loop was previously described to be disordered in the wild-type apo-streptavidin and to close over the pocket upon complexation of the natural ligand biotin. Our findings suggest that stabilization of the 'open' loop conformation in the absence of a ligand abolishes the need for conformational rearrangement prior to the docking of the voluminous peptide. Because no direct contacts between the flexible part of the loop and the peptide ligand were detected, it seems likely that the higher affinity of the two streptavidin mutants for the Strep-tag II is caused by a predominantly entropic mechanism. PMID- 11910033 TI - Alkaline phosphatase from the hyperthermophilic bacterium T. maritima requires cobalt for activity. AB - The hyperthermophilic bacterium Thermotoga maritima encodes a gene sharing sequence similarities with several known genes for alkaline phosphatase (AP). The putative gene was isolated and the corresponding protein expressed in Escherichia coli, with and without a predicted signal sequence. The recombinant protein showed phosphatase activity toward the substrate p-nitrophenyl-phosphate with a k(cat) of 16 s(-1) and a K(m) of 175 microM at a pH optimum of 8.0 when assayed at 25 degrees C. T. maritima phosphatase activity increased at high temperatures, reaching a maximum k(cat) of 100 s(-1), with a K(m) of 93 microM at 65 degrees C. Activity was stable at 65 degrees C for >24 h and at 90 degrees C for 5 h. Phosphatase activity was dependent on divalent metal ions, specifically Co(II) and Mg(II). Circular dichroism spectra showed that the enzyme gains secondary structure on addition of these metals. Zinc, the most common divalent metal ion required for activity in known APs, was shown to inhibit the T. maritima phosphatase enzyme at concentrations above 0.3 moles Zn: 1 mole monomer. All activity was abolished in the presence of 0.1 mM EDTA. The T. maritima AP primary sequence is 28% identical when compared with E. coli AP. Based on a structural model, the active sites are superimposable except for two residues near the E. coli AP Mg binding site, D153 and K328 (E. coli numbering) corresponding to histidine and tryptophan in T. maritima AP, respectively. Sucrose-density gradient sedimentation experiments showed that the protein exists in several quaternary forms predominated by an octamer. PMID- 11910032 TI - Subunit composition of a bicomponent toxin: staphylococcal leukocidin forms an octameric transmembrane pore. AB - Staphylococcal leukocidin pores are formed by the obligatory interaction of two distinct polypeptides, one of class F and one of class S, making them unique in the family of beta-barrel pore-forming toxins (beta-PFTs). By contrast, other beta-PFTs form homo-oligomeric pores; for example, the staphylococcal alpha hemolysin (alpha HL) pore is a homoheptamer. Here, we deduce the subunit composition of a leukocidin pore by two independent methods: gel shift electrophoresis and site-specific chemical modification during single-channel recording. Four LukF and four LukS subunits coassemble to form an octamer. This result in part explains properties of the leukocidin pore, such as its high conductance compared to the alpha HL pore. It is also pertinent to the mechanism of assembly of beta-PFT pores and suggests new possibilities for engineering these proteins. PMID- 11910034 TI - Modeling the transmembrane domain of bacterial chemoreceptors. AB - Bacterial chemoreceptors signal across the membrane by conformational changes that traverse a four-helix transmembrane domain. High-resolution structures are available for the chemoreceptor periplasmic domain and part of the cytoplasmic domain but not for the transmembrane domain. Thus, we constructed molecular models of the transmembrane domains of chemoreceptors Trg and Tar, using coordinates of an unrelated four-helix coiled coil as a template and the X-ray structure of a chemoreceptor periplasmic domain to establish register and positioning. We tested the models using the extensive data for cross-linking propensities between cysteines introduced into adjacent transmembrane helices, and we found that many aspects of the models corresponded with experimental observations. The one striking disparity, the register of transmembrane helix 2 (TM2) relative to its partner transmembrane helix 1, could be corrected by sliding TM2 along its long axis toward the periplasm. The correction implied that axial sliding of TM2, the signaling movement indicated by a large body of data, was of greater magnitude than previously thought. The refined models were used to assess effects of inter-helical disulfides on the two ligand-induced conformational changes observed in alternative crystal structures of periplasmic domains: axial sliding within a subunit and subunit rotation. Analyses using a measure of disulfide potential energy provided strong support for the helical sliding model of transmembrane signaling but indicated that subunit rotation could be involved in other ligand-induced effects. Those analyses plus modeled distances between diagnostic cysteine pairs indicated a magnitude for TM2 sliding in transmembrane signaling of several angstroms. PMID- 11910035 TI - Thermodynamics of single peptide bond cleavage in bovine pancreatic trypsin inhibitor (BPTI). AB - A major goal of this paper was to estimate a dynamic range of equilibrium constant for the opening of a single peptide bond in a model protein, bovine pancreatic trypsin inhibitor (BPTI). Ten mutants of BPTI containing a single Xaa- >Met substitution introduced in different parts of the molecule were expressed in Escherichia coli. The mutants were folded, purified to homogeneity, and cleaved with cyanogen bromide to respective cleaved forms. Conformation of the intact mutants was similar to the wildtype, as judged from their circular dichroism spectra. Substantial conformational changes were observed on the chemical cleavage of three single peptide bonds--Met46-Ser, Met49-Cys, and Met53-Thr- located within the C-terminal helix. Cleavage of those peptide bonds caused a significant destabilization of the molecule, with a drop of the denaturation temperature by 56.4 degrees C to 68 degrees C at pH 4.3. Opening of the remaining seven peptide bonds was related to a 10.8 degrees C to 39.4 degrees C decrease in T(den). Free energies of the opening of 10 single peptide bonds in native mutants (Delta G(op,N)) were estimated from the thermodynamic cycle that links denaturation and cleavage free energies. To calculate those values, we assumed that the free energy of opening of a single peptide bond in the denatured state (Delta G(op,D)) was equal to -2.7 kcal/mole, as reported previously. Calculated Delta G(op,N) values in BPTI were in the range from 0.2 to 10 kcal/mole, which was equivalent to a >1 million-fold difference in equilibrium constants. The values of Delta G(op,N) were the largest for peptide bonds located in the C terminal helix and significantly lower for peptide bonds in the beta-structure or loop regions. It appears that opening constants for single peptide bonds in various proteins span across 33 orders of magnitude. Typical equilibrium values for a single peptide bond opening in a protein containing secondary structure elements fall into negligibly low values, from 10(-3) to 10(-8), and are efficient to ensure stability against proteolysis. PMID- 11910036 TI - Free-thiol Cys331 exposed during activation process is critical for native tetramer structure of cathepsin C (dipeptidyl peptidase I). AB - The mature bovine cathepsin C (CC) molecule is composed of four identical monomers, each proteolytically processed into three chains. Five intrachain disulfides and three nonpaired cysteine residues per monomer were identified. Beside catalytic Cys234 in the active site, free-thiol Cys331 and Cys424 were characterized. Cys424 can be classified as inaccessible buried residue. Selective modification of Cys331 results in dissociation of native CC tetramer into dimers. The 3D homology-based model of the CC catalytic core suggests that Cys331 becomes exposed as the activation peptide is removed during procathepsin C activation. The model further shows that exposed Cys331 is surrounded by a surface hydrophobic cluster, unique to CC, forming a dimer-dimer interaction interface. Substrate/inhibitor recognition of the active site in the CC dimer differs significantly from that in the native tetramer. Taken together, a mechanism is proposed that assumes that the CC tetramer formation results in a site-specific occlusion of endopeptidase-like active site cleft of each CC monomeric unit. Thus, tetramerization provides for the structural basis of the dipeptidyl peptidase activity of CC through a substrate access-limiting mechanism different from those found in homologous monomeric exopeptidases cathepsin H and B. In conclusion, the mechanism of tetramer formation as well as specific posttranslational processing segregates CC in the family of papain proteases. PMID- 11910037 TI - Ca(II)- and Tb(III)-induced stabilization and refolding of anticoagulation factor I from the venom of Agkistrodon acutus. AB - Anticoagulation factor I (ACF I) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X-binding protein in a Ca(2+)-dependent fashion with marked anticoagulant activity. The equilibrium unfolding/refolding of apo ACF I, holo-ACF I, and Tb(3+)-reconstituted ACF I in guanidine hydrochloride (GdnHCl) solutions was studied by following the fluorescence and circular dichroism. Metal ions were found to increase the structural stability of ACF I against GdnHCl and thermal denaturation and, furthermore, influence its unfolding/refolding behavior. The GdnHCl-induced unfolding/refolding of both apo ACF I and Tb(3+)-ACF I is a two-state process with no detectable intermediate state(s), whereas the GdnHCl-induced unfolding/refolding of holo-ACF I in the presence of 1 mM Ca(2+) follows a three-step transition, with intermediate state a (Ia) and intermediate state b (Ib). Ca(2+) ions play an important role in the stabilization of the Ia and Ib states. The decalcification of holo-ACF I shifts the ending zone of unfolding/refolding curve toward lower GdnHCl concentration, whereas the reconstitution of apo-ACF I with Tb(3+) ions shifts the initial zone of denaturation curve toward higher GdnHCl concentration. Therefore, it is possible to find a denaturant concentration (2.0 M GdnHCl) at which refolding from the fully denatured state of apo-ACF I to the Ib state of holo-ACF I or to the native state of Tb(3+)-ACF I can be initiated merely by adding the 1 mM Ca(2+) ions or 10 microM Tb(3+) ions to the unfolded state of apo-ACF I, respectively, without changing the concentration of the denaturant. Using Tb(3+) as a fluorescence probe of Ca(2+), the kinetic results of metal ions-induced refolding provide evidence that the compact Tb(3+)-binding region forms first, and subsequently, the protein undergoes further conformational rearrangements to form the native structure. PMID- 11910038 TI - Dynamical characterization of residual and non-native structures in a partially folded protein by (15)N NMR relaxation using a model based on a distribution of correlation times. AB - A spectral density model based on a truncated lorentzian distribution of correlation times is used to analyze the nanosecond time-scale dynamics of the partially unfolded domain 2 of annexin I from its (15)N NMR relaxation parameters measured at three magnetic field strengths. The use of a distribution of correlation times enables the characterization of the dynamical features of the NH bonds of the protein in terms of heterogeneity of dynamical states in the nanosecond range. The variation along the sequence of the two dynamical parameters introduced, namely the center and the width of the distribution, points out the different types of residual secondary structures present in the D2 domain. Moreover, it allows a physically sensible interpretation of the dynamical behavior of the different residual helices and of the non-native structures. Also, a striking correspondence is found between the parameters obtained using an extended Lipari and Szabo model and the parameters obtained using the distribution of correlation times. This result led us to propose a specific interpretation of the model-free order parameter for internal motions in the nanosecond range in the case of unfolded states. PMID- 11910039 TI - Direct proton magnetic resonance determination of the pKa of the active center histidine in thiolsubtilisin. AB - The serine proteases constitute a group of endopeptidases whose members owe their catalytic activity to the presence of a catalytic triad of amino acids consisting of a serine, a histidine and an aspartate. The pK(a) values for this histidine have been determined for several cases in which there is a negative charge installed at the serine to mimic the oxyanionic intermediate and related transition state for the catalytic pathway. Instances from this laboratory include (1) replacement of the serine by a cysteine in subtilisin to create a thiolate; (2) formation of monoisopropylphosphoryl-Ser 195 monoanionic phosphodiesters (in trypsin and chymotrypsin, Ser 221 in subtilisins); and (3) tetrahedral boronates formed with peptide boronic acids. The nuclear magnetic resonance (NMR) signals pertinent to this histidine, or signals indirectly reflecting the state of ionization of this histidine, have been used effectively to monitor changes in the active center ionization state. In every case studied, there is elevation of the pK(a) at the histidine when the negative charge is installed at the serine position. Herein is reported the first NMR measurement of the active center His 63 pK(a) in thiolsubtilisin Carlsberg; it is elevated by 3 units compared with the parent enzyme. Using a numerical solution (finite difference) of the Poisson-Boltzmann equation, a protein dielectric constant of 4 provides a good estimate of the experimentally observed pK(a) elevations. Very significantly, a very low protein dielectric constant (epsilon(p) = 3-5) is required in all of the comparisons, and for all three enzymes used (chymotrypsin, trypsin, and subtilisin). Finally, we discuss why the electrostatic perturbation sensed at His of the active center is more amplified by a negative charge on the Ser side than the same charge on the Asp side. A plausible explanation is that the positive charge on the imidazolium ring of the His is localized, with the N(delta 1) carrying a smaller fraction, the N(epsilon 2) carrying the bulk of the positive charge. PMID- 11910040 TI - Acyl group specificity at the active site of tetrahydridipicolinate N succinyltransferase. AB - Tetrahydrodipicolinate N-succinyltransferase (DapD) catalyzes the succinyl-CoA dependent acylation of L-2-amino-6-oxopimelate to 2-N-succinyl-6-oxopimelate as part of the succinylase branch of the meso-diaminopimelate/lysine biosynthetic pathway of bacteria, blue-green algae, and plants. This pathway provides meso diaminopimelate as a building block for cell wall peptidoglycan in most bacteria, and is regarded as a target pathway for antibacterial agents. We have solved the X-ray crystal structures of DapD in ternary complexes with pimelate/succinyl-CoA and L-2-aminopimelate with the nonreactive cofactor analog, succinamide-CoA. These structures define the binding conformation of the cofactor succinyl group and its interactions with the enzyme and place its thioester carbonyl carbon in close proximity to the nucleophilic 2-amino group of the acceptor, in support of a direct attack ternary complex mechanism. The acyl group specificity differences between homologous tetrahydrodipicolinate N-acetyl- and N-succinyltransferases can be rationalized with reference to at least three amino acids that interact with or give accessible active site volume to the cofactor succinyl group. These residues account at least in part for the substrate specificity that commits metabolic intermediates to either the succinylase or acetylase branches of the meso-diaminopimelate/lysine biosynthetic pathway. PMID- 11910041 TI - Polyproline II helical structure in protein unfolded states: lysine peptides revisited. AB - The left-handed polyproline II (PPII) helix gives rise to a circular dichroism spectrum that is remarkably similar to that of unfolded proteins. This similarity has been used as the basis for the hypothesis that unfolded proteins possess considerable PPII helical content. It has long been known that homopolymers of lysine adopt the PPII helical conformation at neutral pH, presumably a result of electrostatic repulsion between side chains. It is shown here that a seven residue lysine peptide also adopts the PPII conformation. In contrast with homopolymers of lysine, this short peptide is shown to retain PPII helical character under conditions in which side-chain charges are heavily screened or even neutralized. The most plausible explanation for these observations is that the peptide backbone favors the PPII conformation to maximize favorable interactions with solvent. These data are evidence that unfolded proteins do indeed possess PPII content, indicating that the ensemble of unfolded states is significantly smaller than is commonly assumed. PMID- 11910042 TI - Free energies of protein decoys provide insight into determinants of protein stability. PMID- 11910043 TI - Comments to the editor concerning the paper entitled "Effects of endotoxin on neutrophil-mediated ischemia/reperfusion injury in the rat heart in vivo" by Lipton et al. PMID- 11910046 TI - Control of coronary blood flow during exercise. AB - Under normal physiological conditions, coronary blood flow is closely matched with the rate of myocardial oxygen consumption. This matching of flow and metabolism is physiologically important due to the limited oxygen extraction reserve of the heart. Thus, when myocardial oxygen consumption is increased, as during exercise, coronary vasodilation and increased oxygen delivery are critical to preventing myocardial underperfusion and ischemia. Exercise coronary vasodilation is thought to be mediated primarily by the production of local metabolic vasodilators released from cardiomyocytes secondary to an increase in myocardial oxygen consumption. However, despite various investigations into this mechanism, the mediator(s) of metabolic coronary vasodilation remain unknown. As will be seen in this review, the adenosine, K(+)(ATP) channel and nitric oxide hypotheses have been found to be inadequate, either alone or in combination as multiple redundant compensatory mechanisms. Prostaglandins and potassium are also not important in steady-state coronary flow regulation. Other factors such as ATP and endothelium-derived hyperpolarizing factors have been proposed as potential local metabolic factors, but have not been examined during exercise coronary vasodilation. In contrast, norepinephrine released from sympathetic nerve endings mediates a feed-forward betaadrenoceptor coronary vasodilation that accounts for approximately 25% of coronary vasodilation observed during exercise. There is also a feed-forward alpha-adrenoceptor-mediated vasoconstriction that helps maintain blood flow to the vulnerable subendocardium when heart rate, myocardial contractility, and oxygen consumption are elevated during exercise. Control of coronary blood flow during pathophysiological conditions such as hypertension, diabetes mellitus, and heart failure is also addressed. PMID- 11910045 TI - The potential of DNA vaccination against tumor-associated antigens for antitumor therapy. AB - Conventional treatment approaches for malignant tumors are highly invasive and sometimes have only a palliative effect. Therefore, there is an increasing demand to develop novel, more efficient treatment options. Increased efforts have been made to apply immunomodulatory strategies in antitumor treatment. In recent years, immunizations with naked plasmid DNA encoding tumor-associated antigens have revealed a number of advantages. By DNA vaccination, antigen-specific cellular as well as humoral immune responses can be generated. The induction of specific immune responses directed against antigens expressed in tumor cells and displayed e.g., by MHC class I complexes can inhibit tumor growth and lead to tumor rejection. The improvement of vaccine efficacy has become a critical goal in the development of DNA vaccination as antitumor therapy. The use of different DNA delivery techniques and coadministration of adjuvants including cytokine genes may influence the pattern of specific immune responses induced. This brief review describes recent developments to optimize DNA vaccination against tumor associated antigens. The prerequisite for a successful antitumor vaccination is breaking tolerance to tumor-associated antigens, which represent "self-antigens." Currently, immunization with xenogeneic DNA to induce immune responses against self-molecules is under intensive investigation. Tumor cells can develop immune escape mechanisms by generation of antigen loss variants, therefore, it may be necessary that DNA vaccines contain more than one tumor antigen. Polyimmunization with a mixture of tumor-associated antigen genes may have a synergistic effect in tumor treatment. The identification of tumor antigens that may serve as targets for DNA immunization has proceeded rapidly. Preclinical studies in animal models are promising that DNA immunization is a potent strategy for mediating antitumor effects in vivo. Thus, DNA vaccines may offer a novel treatment for tumor patients. DNA vaccines may also be useful in the prevention of tumors with genetic predisposition. By DNA vaccination preventing infections, the development of viral-induced tumors may be avoided. PMID- 11910047 TI - Systolic elastance and resistance in the regulation of cardiac pumping function in early streptozotocin-diabetic rats. AB - We determined the roles of maximal systolic elastance (E(max)) and theoretical maximum flow ((max)) in the regulation of cardiac pumping function in early streptozotocin (STZ)-diabetic rats. Physically, E(max) can reflect the intrinsic contractility of the myocardium as an intact heart, and (max) has an inverse relation to the systolic resistance of the left ventricle. Rats given STZ 65 mg/kg i.v. (n = 17) were divided into two groups, 1 week and 4 weeks after induction of diabetes, and compared with untreated age-matched controls (n = 15). Left ventricular (LV) pressure and ascending aortic flow signals were recorded to calculate E(max) and (max), using the elastance-resistance model. After 1 or 4 weeks, STZ-diabetic animals show an increase in effective LV end-diastolic volume (V(eed)), no significant change in peak isovolumic pressure (P(iso)(max)), and a decline in effective arterial volume elastance (E(a)). The maximal systolic elastance E(max) is reduced from 751.5 +/- 23.1 mmHg/ml in controls to 514.1 +/- 22.4 mmHg/ml in 1- and 538.4 +/- 33.8 mmHg/ml in 4-week diabetic rats. Since E(max) equals P(iso)(max)/V(eed), an increase in V(eed) with unaltered P(iso)(max) may primarily act to diminish E(max) so that the intrinsic contractility of the diabetic heart is impaired. By contrast, STZ-diabetic rats have higher theoretical maximum flow (max) (40.9 +/- 2.8 ml/s in 1- and 44.5 +/- 3.8 ml/s in 4-week diabetic rats) than do controls (30.7 +/- 1.7 ml/s). There exists an inverse relation between (max) and E(a) when a linear regression of (max) on E(a) is performed over all animals studied (r = 0.65, p < 0.01). The enhanced (max) is indicative of the decline in systolic resistance of the diabetic rat heart. The opposing effects of enhanced (max) and reduced E(max) may negate each other, and then the cardiac pumping function of the early STZ diabetic rat heart could be preserved before cardiac failure occurs. PMID- 11910048 TI - Ethanol modulates the growth of human breast cancer cells in vitro. AB - The role of ethanol or its metabolites on breast neoplasm has not been characterized. We hypothesized that ethanol may alter the growth rate of human breast tumor epithelial cells by modulating putative growth-promoting signaling pathways such as p44/42 mitogen-activated protein kinases (MAPKs). The MCF-7 cell line, considered a suitable model, was used in these studies to investigate the effects of ethanol on [(3)H]thymidine incorporation, cell number, and p44/42 MAPK activities in the presence or absence of a MAPK or extracellular signal-regulated kinase ERK-1, and (MEK1) inhibitor (PD098059). Treatment of MCF-7 cells with a physiologically relevant concentration of ethanol (0.3% or 65 mM) increased p44/42 activities by an average of 400% (P < 0.02), and subsequent cell growth by 200% (P < 0.05) in a MEK1 inhibitor (PD098059)-sensitive fashion, thus suggesting that the Ras/MEK/MAPK signaling pathways are crucial for ethanol-induced MCF-7 cell growth. PMID- 11910049 TI - Increased beef consumption increases apolipoprotein A-I but not serum cholesterol of mildly hypercholesterolemic men with different levels of habitual beef intake. AB - The objective of this research was to compare the effects of a lean beef enriched in oleic acid to a beef that is typical of the commercial beef consumed in the United States. Ten mildly hypercholesterolemic men, ages 34-58 years old, were selected from the Texas A&M University faculty and staff. Subjects were randomly assigned to one of two diets for a 6-week duration followed by a crossover after a 4-week habitual diet washout period. Diets were consumed daily for a 6-week study period. Participants substituted lean beef obtained from Wagyu bullocks or commercial beef for the meat typically consumed. Total cholesterol, apolipoproteins A-I and B, triacylglycerols, and low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol were measured in serum samples collected weekly. Beef type had no effect on any measured variable. There were no significant differences between baseline HDL or LDL cholesterol concentrations after the consumption of the beef test diets. Apolipoprotein A-I, serum glucose, and uric acid concentrations were elevated by the additional dietary beef. Analysis of records of customary diets indicated that one group consumed 160 g of beef daily, whereas the other group consumed only 26 g of beef daily. Therefore, post hoc analyses tested the habitual beef intake x treatment time interaction. LDL cholesterol concentration was markedly higher in the group with low habitual beef intake (180 vs 144 mg/dl), and HDL cholesterol was slightly higher (44 vs 40 mg/dl; post-test values) than for the group with high habitual beef intake, but there were no habitual intake x time interactions for LDL or HDL cholesterol. Creatinine and blood urea nitrogen concentrations also were greater in the individuals habitually consuming less beef. This study had three important findings: i) a lean beef source enriched with oleic acid was no different from commercial beef in its effect on lipoprotein fractions; ii) neither previous level of beef intake nor baseline LDL cholesterol concentration influenced the serum cholesterol response to added dietary beef, which was negative; and iii) apolipoprotein A-I, but not HDL or LDL cholesterol, was sensitive to the additional dietary beef. PMID- 11910050 TI - Effects of polyamines on the release of gonadotropin-releasing hormone and gonadotropins in developing female rats. AB - Polyamines, putrescine (PUT), spermidine (SPD), spermine (SPM), and agmatine (AGM), are polycationic amines related to multiple cell functions found in high concentrations during the development of hypothalamus and pituitary. In previous works, we demonstrated that alpha-difluoromethylornithine (DFMO), an inhibitor of polyamines biosynthesis, induced a delay in puberty of female rats, accompanied by high, sustained follicle-stimulating hormone (FSH) levels during the infantile period. Also, DFMO treatment induced changes in polyamine concentration both in hypothalamus and pituitary of rats, mainly a decrease of PUT and SPD, an increase in SPM, and no change in AGM. In the present work, we investigated the direct effects of polyamines on the secretion of hypothalamic GnRH and pituitary gonadotropins in 6- and 15-day-old female rats. In 6-day-old animals, in vitro incubations with PUT, SPD, and AGM of hypothalami or anterior pituitaries were able to inhibit GnRH, FSH, and leutinizing hormone (LH) secretion, respectively. SPM showed a nonspecific transient inhibitory effect on FSH. When challenged with either high K(+) (hypothami) or GnRH (pituitaries), the tissues incubated in the presence of polyamines showed no differences when compared with their controls. No effects of polyamines in 15-day-old rats in either tissue were observed. Pituitary cell cultures of 6-day-old animals incubated with DFMO for 4 days showed a significant increase in FSH, but not in LH. We conclude that high PUT, SPD, and AGM levels during the first 10 days of life are important for the development of the hypothalamic-hypophyseal unit, probably related to an inhibitory effect on GnRH and gonadotropins. Therefore, polyamine participation, especially PUT and SPD, is of importance in the regulation of GnRH and gonadotropin secretion in the neonatal and infantile periods, critical stages in the establishment of sexual differentiation. PMID- 11910051 TI - Effect of modified diabetic splenocytes on mice injected with multiple low-dose streptozotocin. AB - This work reports the effects of a previous injection of mitomycin-modified splenocytes from multiple-low dose streptozotocin-treated mice (mld-sz) on autoimmune diabetes produced by mld-sz. Our work shows that a previous inoculation of modified mononuclear splenocytes from mld-sz mice prevents alterations in glycemia, in insulin secretion (IS) pattern from isolated perifused islets, and in mass of pancreatic islets. Immunohistochemistry showed an alteration in the number of beta, but not of alpha or delta cells. While a mononuclear intra-islet infiltration was observed in mld-sz mice, a predominantly polar or peri-islet infiltration was seen in vaccinated mice. Islet-associated mononuclear cells from mld-sz mice produced diabetes and induced a diminished IS when transferred to normal receptors. Those cells from previously vaccinated mld sz mice had no effect when injected into normal receptors. In addition, they also inhibited the damage induced in normal receptors by the islet-associated mononuclear cells from mld-sz animals. Cellular death was also prevented by previous vaccination. Our results suggest that vaccination with modified splenocytes from mld-sz mice is capable of shifting the islet cells infiltration pattern from an aggressive one toward a protective one and thus preventing the beta cell destruction observed in mld-sz mice. PMID- 11910052 TI - Public health quality measurement: concepts and challenges. AB - Public health agencies increasingly are recognizing the need to formally and quantitatively assess and improve the quality of their programs, information, and policies. Measuring quality can help organizations monitor their progress toward public health goals and become more accountable to both the populations they serve and policy makers. Yet quality assessment is a complex task that involves precise determination and specification of useful measures. We discuss a well established conceptual framework for organizing quality assessment in the context of planning and delivery of programs and services by local health departments, and consider the strengths and limitations of this approach for guiding quality improvement. We review several past and present quality measurement-related initiatives designed for public health department use, and discuss current and future challenges in this evolving area of public health practice. PMID- 11910053 TI - Cascade effects of medical technology. AB - Cascade effect refers to a process that proceeds in stepwise fashion from an initiating event to a seemingly inevitable conclusion. With regard to medical technology, the term refers to a chain of events initiated by an unnecessary test, an unexpected result, or patient or physician anxiety, which results in ill advised tests or treatments that may cause avoidable adverse effects and/or morbidity. Examples include discovery of endocrine incidentalomas on head and body scans; irrelevant abnormalities on spinal imaging; tampering with random fluctuations in clinical measures; and unwanted aggressive care at the end of life. Common triggers include failing to understand the likelihood of false positive results; errors in data interpretation; overestimating benefits or underestimating risks; and low tolerance of ambiguity. Excess capacity and perverse financial incentives may contribute to cascade effects as well. Preventing cascade effects may require better education of physicians and patients; research on the natural history of mild diagnostic abnormalities; achieving optimal capacity in health care systems; and awareness that more is not the same as better. PMID- 11910054 TI - The effectiveness of state-level tobacco control interventions: a review of program implementation and behavioral outcomes. AB - In 2001, nearly one billion dollars will be spent on statewide tobacco control programs, including those in California, Massachusetts, Arizona, and Oregon, funded by cigarette tax revenues, and the program in Florida, funded by the state's settlement with the tobacco industry. With such large expenditures, it is imperative to find out whether these programs are working. This paper reviews the effectiveness of the statewide tobacco control programs in California, Massachusetts, Arizona, Oregon, and Florida. It focuses on two aspects of process evaluation--the funding and implementation of the programs and the tobacco industry's response, and four elements of outcome evaluation--the programs' effects on cigarette consumption, adult and youth smoking prevalence, and protection of the public from secondhand smoke. The paper formulates general lessons learned from these existing programs and generates recommendations to improve and inform the development and implementation of these and future programs. PMID- 11910055 TI - Direct marketing of pharmaceuticals to consumers. AB - Revised FDA regulations governing pharmaceutical companies' broadcast advertisements directed to consumers produced substantial increases in direct-to consumer advertising (DTCA) expenditures. Proponents of DTCA claim it supports patient autonomy in the patient-physician relationship and has motivated some consumers to seek a physician's care for conditions they previously had not discussed with a doctor. However, DTCA's blend of promotion and information has produced more prescription drug awareness than knowledge--it has been largely ineffective in educating patients with medical conditions about the medications for those conditions. The evidence for DTCA's increase in pharmaceutical sales is as impressive as is the lack of evidence concerning its impact on the health of the public. Broadcast advertisements are too brief to include extensive technical information; consequently, the impact of FDA regulations to assure a fair balance of risk and benefit in DTCA is still being assessed. PMID- 11910056 TI - Challenges in motor vehicle safety. AB - Reductions in motor vehicle injury and death represent a major public health success. Since the advent of the federal program in highway safety in 1966, motor vehicle deaths have dropped dramatically, not only in rates per miles driven but also in absolute numbers. Key to this success has been the broad-based comprehensive approach promoted by the program's first administrator, a public health physician. The federal program provided leadership and coordination that leveraged national, state, and local programs to bring about safer vehicles, improved traffic records, more effective enforcement, enormously improved emergency medical services, more responsible judicial systems, and many other accomplishments. Although progress has been made on many fronts, major areas addressed here include federal motor vehicle safety standards, alcohol safety programs, occupant restraint laws and usage, and speed limits. The achievements in motor vehicle safety provide a model for other injury control efforts. PMID- 11910057 TI - HALYS and QALYS and DALYS, Oh My: similarities and differences in summary measures of population Health. AB - Health-adjusted life years (HALYs) are population health measures permitting morbidity and mortality to be simultaneously described within a single number. They are useful for overall estimates of burden of disease, comparisons of the relative impact of specific illnesses and conditions on communities, and in economic analyses. Quality-adjusted life years (QALYs) and disability-adjusted life years (DALYs) are types of HALYs whose original purposes were at variance. Their growing importance and the varied uptake of the methodology by different U.S. and international entities makes it useful to understand their differences as well as their similarities. A brief history of both measures is presented and methods for calculating them are reviewed. Methodological and ethical issues that have been raised in association with HALYs more generally are presented. Finally, we raise concerns about the practice of using different types of HALYs within different decision-making contexts and urge action that builds and clarifies this useful measurement field. PMID- 11910058 TI - Morbidity and mortality from medical errors: an increasingly serious public health problem. AB - From 1983 to 1998, U.S. fatalities from acknowledged prescription errors increased by 243%, from 2,876 to 9,856. This percentage increase was greater than for almost any other cause of death, and far outpaced the increase in the number of prescriptions. Many nonfatal prescription errors also occur, but estimates of the frequency of these errors vary widely, because various definitions, geographic settings, and institutions have been used. Efforts to reduce fatal and nonfatal prescription errors have encountered perceptual, legal, medical, and cultural barriers. It may be possible to reduce prescription errors by instituting a central agency responsible for collecting, analyzing, and reporting harmful or potentially harmful drug events, and for issuing recommendations and directives. PMID- 11910059 TI - The importance of the normality assumption in large public health data sets. AB - It is widely but incorrectly believed that the t-test and linear regression are valid only for Normally distributed outcomes. The t-test and linear regression compare the mean of an outcome variable for different subjects. While these are valid even in very small samples if the outcome variable is Normally distributed, their major usefulness comes from the fact that in large samples they are valid for any distribution. We demonstrate this validity by simulation in extremely non Normal data. We discuss situations in which in other methods such as the Wilcoxon rank sum test and ordinal logistic regression (proportional odds model) have been recommended, and conclude that the t-test and linear regression often provide a convenient and practical alternative. The major limitation on the t-test and linear regression for inference about associations is not a distributional one, but whether detecting and estimating a difference in the mean of the outcome answers the scientific question at hand. PMID- 11910061 TI - The public health impact of Alzheimer's disease, 2000-2050: potential implication of treatment advances. AB - Recent developments in basic research suggest that therapeutic breakthroughs may occur in Alzheimer's disease treatment over the coming decades. To model the potential magnitude and nature of the effect of these advances, historical data from congestive heart failure and Parkinson's disease were used. Projections indicate that therapies which delay disease onset will markedly reduce overall disease prevalence, whereas therapies to treat existing disease will alter the proportion of cases that are mild as opposed to moderate/severe. The public health impact of such changes would likely involve both the amount and type of health services needed. Particularly likely to arise are new forms of outpatient services, such as disease-specific clinics and centers. None of our models predicts less than a threefold rise in the total number of persons with Alzheimer's disease between 2000 and 2050. Therefore, Alzheimer's care is likely to remain a major public health problem during the coming decades. PMID- 11910060 TI - Gambling and related mental disorders: a public health analysis. AB - This article reviews the prevalence of gambling and related mental disorders from a public health perspective. It traces the expansion of gambling in North America and the psychological, economic, and social consequences for the public's health, and then considers both the costs and benefits of gambling and the history of gambling prevalence research. A public health approach is applied to understanding the epidemiology of gambling-related problems. International prevalence rates are provided and the prevalence of mental disorders that often are comorbid with gambling problems is reviewed. Analysis includes an examination of groups vulnerable to gambling-related disorders and the methodological and conceptual matters that might influence epidemiological research and prevalence rates related to gambling. The major public health problems associated with gambling are considered and recommendations made for public health policy, practice, and research. The enduring value of a public health perspective is that it applies different 'lenses' for understanding gambling behaviour, analysing its benefits and costs, as well as identifying strategies for action. Harvey A. Skinner (160, p. 286) PMID- 11910062 TI - Utilization management: issues, effects, and future prospects. AB - Utilization management encompasses a diverse set of activities designed to influence the use of health care services and thereby constrain health care resource consumption. Utilization management, which has become one of the most widely used cost-containment approaches, has engendered debate and controversy. Physicians have been outspoken critics of utilization management because it has limited their clinical autonomy and has contributed to an intolerable administrative burden. Insurance carriers, managed care plans, and third-party payers have defended the use of utilization management as an imperfect-but necessary-practice that is needed to reduce consumption of unnecessary or inappropriate health care services. This review examines the operation and effects of three widely used utilization management procedures: prospective utilization review, case management, and physician gatekeeping programs. In addition, it explores the future role of utilization management in the health care system and outlines a set of principles that we believe should be used to guide the development of utilization management strategies in the future. PMID- 11910063 TI - Dietary interventions to prevent disease. AB - Changing dietary behaviors to prevent chronic disease has been an important research focus for the last 25 years. Here we present a review of published articles on the results of research to identify methods to change key dietary habits: fat intake, fiber intake, and consumption of fruits and vegetables. We divided the research reviewed into sections, based on the channel through which the intervention activities were delivered. We conclude that the field is making progress in identifying successful dietary change strategies, but that more can be learned. Particularly, we need to transfer some of the knowledge from the individual-based trials to community-level interventions. Also, more research with rigorous methodology must be done to test current and future intervention options. PMID- 11910064 TI - The macroeconomic determinants of health. AB - Why are some societies healthier than others? The consensus in development economics is that the health achievement of nations has to do with their levels of economic development. Higher per capita incomes, through steady and stable economic growth, increase a nation's capacity to purchase the necessary economic goods and services that promote health. In this paper, we review the conceptual and empirical linkages between poverty and poor health in both developing and developed countries. The empirical evidence is overwhelming that poverty, measured at the level of societies as well as individuals, is causally related to poor health of societies and individuals, respectively. Recent macroeconomic research has also drawn attention to the role of health as a form of human capital that is vital for achieving economic stability. In particular, attention has been drawn toward the ways in which unhealthy societies impede the process of economic development. However, the reciprocal connection between economic prosperity and improved health is neither automatic nor universal. Other features of society, such as the equality in the distribution of the national wealth, seem to matter as well for improving average population health and especially for reducing inequalities in health. We conclude by arguing for a need to reexamine the way in which health is conceptualized within the macroeconomic development framework. PMID- 11910065 TI - Socioeconomic status and health: the potential role of environmental risk exposure. AB - Among several viable explanations for the ubiquitous SES-health gradient is differential exposure to environmental risk. We document evidence of inverse relations between income and other indices of SES with environmental risk factors including hazardous wastes and other toxins, ambient and indoor air pollutants, water quality, ambient noise, residential crowding, housing quality, educational facilities, work environments, and neighborhood conditions. We then briefly overview evidence that such exposures are inimical to health and well-being. We conclude with a discussion of the research and policy implications of environmental justice, arguing that a particularly salient feature of poverty for health consequences is exposure to multiple environmental risk factors. PMID- 11910066 TI - Effects of smoking restrictions in the workplace. AB - The health hazards caused by exposure to environmental tobacco smoke (ETS) are well established. Workplace exposure to ETS is strongly influenced by the types of workplace and smoking policy-total bans on smoking have become common in many countries. Blue-collar and service workers are more likely than other types of workers to be exposed to ETS in the workplace. Smokers who are employed in workplaces with smoking bans are likely to consume fewer cigarettes per day, are more likely to be considering quitting, and quit at an increased rate compared with smokers employed in workplaces with no or weaker policies. Despite substantial progress in protecting workers from ETS, additional efforts are needed in areas that include attention to exposure among blue-collar and service workers; policies in workplaces with a limited number of employees; and studies of enforcement, effects on smoking cessation in multiple settings, and cost effectiveness. PMID- 11910067 TI - Socioeconomic inequalities in injury: critical issues in design and analysis. AB - Injuries continue to place a tremendous burden on the public's health and rates vary widely among different groups in the population. Increasing attention has recently been given to the effects of socioeconomic status (SES) as a determinant of health among both individuals and communities. However, relatively few studies have focused on the influence of SES and injuries. Furthermore, those that have, and the other injury studies that have included measures of SES in their analysis, have varying degrees of conceptual and methodological rigor in their use of this measure. Recent advances in data linkage and analytic techniques have, however, provided new and improved methods to assess the relationship between SES and injuries. This review summarizes the relevant literature on SES and injuries, with particular attention to study design, and the measurement and interpretation of SES. We found that increasing SES has a strong inverse association with the risk of both homicide and fatal unintentional injuries, although the results for suicide were mixed. However, the relationship between SES and nonfatal injuries was less consistent than for fatal injuries. We offer potential explanatory mechanisms for the relationship between SES and injuries and make recommendations for future research in this area. PMID- 11910068 TI - Thinking outside the box: recent advances in the analysis and presentation of uncertainty in cost-effectiveness studies. AB - As many more clinical trials collect economic information within their study design, so health economics analysts are increasingly working with patient-level data on both costs and effects. In this paper, we review recent advances in the use of statistical methods for economic analysis of information collected alongside clinical trials. In particular, we focus on the handling and presentation of uncertainty, including the importance of estimation rather than hypothesis testing, the use of the net-benefit statistic, and the presentation of cost-effectiveness acceptability curves. We also discuss the appropriate sample size calculations for cost-effectiveness analysis at the design stage of a study. Finally, we outline some of the challenges for future research in this area particularly in relation to the appropriate use of Bayesian methods and methods for analyzing costs that are typically skewed and often incomplete. PMID- 11910069 TI - Tuberculosis. AB - Tuberculosis is an infectious disease caused by bacteria in the Mycobacterium tuberculosis complex. Of these, the most common species to infect humans is M. tuberculosis. The TB bacillus is an extremely successful human pathogen, infecting two billion persons worldwide; an estimated 2 to 3 million people die from tuberculosis each year. In the United States, TB rates decreased steadily at the rate of 5% per year from 1953 until 1985 when the trend reversed, with the number of TB cases peaking in 1992. Outbreaks of multidrug-resistant TB (MDR TB) were reported, and these cases were documented to be transmitted in nosocomial and congregate settings, including hospitals and prisons. AIDS patients infected with M. tb developed disease rapidly, and case-fatality rates of >80% were noted in those infected with multidrug-resistant M. tb. Intensive intervention, at enormous cost, caused the number of TB cases to decline. This article discusses factors that led to the increase in TB cases, their subsequent decline, and measures needed in the future if TB is to be eliminated in the United States. PMID- 11910070 TI - The future of benefit-cost analyses of the Clean Air Act. AB - This review examines the first two studies conducted pursuant to a Congressional mandate that the U.S. Environmental Protection Agency analyze the effects of the Clean Air Act on the "public health, economy, and the environment of the United States." While these studies indicate that overall, the nation received good value for the resources it invested in improving air quality over the past three decades, we don't know if even higher value could have been obtained by changing or eliminating certain potentially inefficient elements. The review focuses on the critical policy and technical choices made in the analyses, including the selection of the appropriate baseline and the level of disaggregation for the studies. It is proposed that a potential third analysis focus on potential new policies not yet mandated by law or regulation. It is also proposed that the next study fill in key information gaps, expand the benefit categories, and incorporate new research on topics such as mortality and morbidity benefits, cost uncertainties, and others. PMID- 11910071 TI - Ordering in a fluid inert gas confined by flat surfaces. AB - High-resolution transmission electron microscopy images of room-temperature fluid xenon in small faceted cavities in aluminum reveal the presence of three well defined layers within the fluid at each facet. Such interfacial layering of simple liquids has been theoretically predicted, but observational evidence has been ambiguous. Molecular dynamics simulations indicate that the density variation induced by the layering will cause xenon, confined to an approximately cubic cavity of volume approximately 8 cubic nanometers, to condense into the body-centered cubic phase, differing from the face-centered cubic phase of both bulk solid xenon and solid xenon confined in somewhat larger (>/=20 cubic nanometer) tetradecahedral cavities in face-centered cubic metals. Layering at the liquid-solid interface plays an important role in determining physical properties as diverse as the rheological behavior of two-dimensionally confined liquids and the dynamics of crystal growth. PMID- 11910072 TI - A system for stable expression of short interfering RNAs in mammalian cells. AB - Mammalian genetic approaches to study gene function have been hampered by the lack of tools to generate stable loss-of-function phenotypes efficiently. We report here a new vector system, named pSUPER, which directs the synthesis of small interfering RNAs (siRNAs) in mammalian cells. We show that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes. Stable expression of siRNAs using this vector mediates persistent suppression of gene expression, allowing the analysis of loss-of-function phenotypes that develop over longer periods of time. Therefore, the pSUPER vector constitutes a new and powerful system to analyze gene function in a variety of mammalian cell types. PMID- 11910073 TI - Phase of matter. The elusive liquid-solid interface. PMID- 11910075 TI - John Gardner: a salute. PMID- 11910074 TI - Functional annotation of a full-length Arabidopsis cDNA collection. AB - Full-length complementary DNAs (cDNAs) are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. We isolated 155,144 RIKEN Arabidopsis full-length (RAFL) cDNA clones. The 3'-end expressed sequence tags (ESTs) of 155,144 RAFL cDNAs were clustered into 14,668 nonredundant cDNA groups, about 60% of predicted genes. We also obtained 5' ESTs from 14,034 nonredundant cDNA groups and constructed a promoter database. The sequence database of the RAFL cDNAs is useful for promoter analysis and correct annotation of predicted transcription units and gene products. Furthermore, the full-length cDNAs are useful resources for analyses of the expression profiles, functions, and structures of plant proteins. PMID- 11910076 TI - Human rights. Statistical analysis provides key links in Milosevic trial. PMID- 11910077 TI - Archaeology. Dam threatens Iraqi ancient sites. PMID- 11910078 TI - Caspian Sea. Scientists deplore OK for sturgeon catch. PMID- 11910079 TI - Women in academia. Engineers marginalized, MIT report concludes. PMID- 11910080 TI - Paleoanthropology. African skull points to one human ancestor. PMID- 11910082 TI - Astrophysics. Distant galaxy heralds end of Dark Ages. PMID- 11910081 TI - Neuroscience. The good, the bad, and the anterior cingulate. PMID- 11910083 TI - Chemistry. Whisper of magnetism tells molecules apart. PMID- 11910084 TI - Forensic science. Judge reverses decision on fingerprint evidence. PMID- 11910085 TI - Evolutionary biology. Ancient DNA untangles evolutionary paths. PMID- 11910087 TI - Agriculture. Reseeding project offers aid to strapped Afghani farmers. PMID- 11910086 TI - Cancer therapy. Setbacks for endostatin. PMID- 11910088 TI - Invasive species. California tries to rub out the monster of the lagoon. PMID- 11910090 TI - Astronomy. Unusual venture helps make the sky affordable. PMID- 11910089 TI - U.S. demographics. New annual survey brings census into 21st century. PMID- 11910091 TI - Bioengineering. Working outside the protein-synthesis rules. PMID- 11910092 TI - Special essay. The seven pillars of life. PMID- 11910093 TI - Nuclear research. The past and future of University research reactors. PMID- 11910094 TI - Molecular biology. Skiing toward nonstop mRNA decay. PMID- 11910095 TI - Chemical reactions. Steric and solvent effects in ionic reactions. PMID- 11910096 TI - Cosmology. The beginning of time. PMID- 11910097 TI - Superconductivity. The true colors of cuprates. PMID- 11910098 TI - Paleoclimate. Blowing hot and cold. PMID- 11910100 TI - Ecology. Of predators, prey, and power laws. PMID- 11910099 TI - Signal transduction. Hot and cold TRP ion channels. PMID- 11910101 TI - Pituitary development: regulatory codes in mammalian organogenesis. AB - During mammalian pituitary gland development, distinct cell types emerge from a common primordium. Appearance of specific cell types occurs in response to opposing signaling gradients that emanate from distinct organizing centers. These signals induce expression of interacting transcriptional regulators, including DNA binding-dependent activators and DNA binding-independent transrepressors, in temporally and spatially overlapping patterns. Together they synergistically regulate precursor proliferation and induction of distinct cell types. Terminal cell type differentiation requires selective gene activation strategies and long term active repression, mediated by cell type-specific and promoter-specific recruitment of coregulatory complexes. These mechanisms imply the potential for flexibility in the ultimate identity of differentiated cell types. PMID- 11910102 TI - Self-assembled magnetic matrices for DNA separation chips. PMID- 11910103 TI - Superconductivity-induced transfer of in-plane spectral weight in Bi2Sr2CaCu2O8+delta. AB - Optical data are reported on a spectral weight transfer over a broad frequency range of Bi2Sr2CaCu2O8+delta, when this material became superconducting. Using spectroscopic ellipsometry, we observed the removal of a small amount of spectral weight in a broad frequency band from 10(4) cm(-1) to at least 2 x 10(4) cm(-1), due to the onset of superconductivity. We observed a blue shift of the ab-plane plasma frequency when the material became superconducting, indicating that the spectral weight was transferred to the infrared range. Our observations are in agreement with models in which superconductivity is accompanied by an increased charge carrier spectral weight. The measured spectral weight transfer is large enough to account for the condensation energy in these compounds. PMID- 11910104 TI - Steric effects and solvent effects in ionic reactions. AB - Rates of SN2 reactions of chloride ion with methyl- and tert-butyl-substituted chloroacetonitrile were measured by using Fourier transform-ion cyclotron resonance spectrometry to follow the isotopic exchange reaction. Barrier heights for these reactions indicate that steric effects in the gas phase are diminished relative to apparent steric effects in solution. We attribute the increased barrier in solution to a solvation effect. Monte Carlo simulations done using statistical perturbation theory confirm that steric hindrance to solvation contributes to SN2 barriers in solution. PMID- 11910106 TI - Low-frequency signals in long tree-ring chronologies for reconstructing past temperature variability. AB - Preserving multicentennial climate variability in long tree-ring records is critically important for reconstructing the full range of temperature variability over the past 1000 years. This allows the putative "Medieval Warm Period" (MWP) to be described and to be compared with 20th-century warming in modeling and attribution studies. We demonstrate that carefully selected tree-ring chronologies from 14 sites in the Northern Hemisphere (NH) extratropics can preserve such coherent large-scale, multicentennial temperature trends if proper methods of analysis are used. In addition, we show that the average of these chronologies supports the large-scale occurrence of the MWP over the NH extratropics. PMID- 11910105 TI - Liquid-state NMR and scalar couplings in microtesla magnetic fields. AB - We obtained nuclear magnetic resonance (NMR) spectra of liquids in fields of a few microtesla, using prepolarization in fields of a few millitesla and detection with a dc superconducting quantum interference device (SQUID). Because the sensitivity of the SQUID is frequency independent, we enhanced both signal-to noise ratio and spectral resolution by detecting the NMR signal in extremely low magnetic fields, where the NMR lines become very narrow even for grossly inhomogeneous measurement fields. In the absence of chemical shifts, proton phosphorous scalar (J) couplings have been detected, indicating the presence of specific covalent bonds. This observation opens the possibility for "pure J spectroscopy" as a diagnostic tool for the detection of molecules in low magnetic fields. PMID- 11910107 TI - Coda wave interferometry for estimating nonlinear behavior in seismic velocity. AB - In coda wave interferometry, one records multiply scattered waves at a limited number of receivers to infer changes in the medium over time. With this technique, we have determined the nonlinear dependence of the seismic velocity in granite on temperature and the associated acoustic emissions. This technique can be used in warning mode, to detect the presence of temporal changes in the medium, or in diagnostic mode, where the temporal change in the medium is quantified. PMID- 11910108 TI - Adaptive immune response of Vgamma2Vdelta2+ T cells during mycobacterial infections. AB - To examine the role of T cell receptor (TCR) in gammadelta T cells in adaptive immunity, a macaque model was used to follow Vgamma2Vdelta2+ T cell responses to mycobacterial infections. These phosphoantigen-specific gammadelta T cells displayed major expansion during Mycobacterium bovis Bacille Calmette-Guerin (BCG) infection and a clear memory-type response after BCG reinfection. Primary and recall expansions of Vgamma2Vdelta2+ T cells were also seen during Mycobacterium tuberculosis infection of naive and BCG-vaccinated macaques, respectively. This capacity to rapidly expand coincided with a clearance of BCG bacteremia and immunity to fatal tuberculosis in BCG-vaccinated macaques. Thus, Vgamma2Vdelta2+ T cells may contribute to adaptive immunity to mycobacterial infections. PMID- 11910109 TI - An mRNA surveillance mechanism that eliminates transcripts lacking termination codons. AB - Translation is an important mechanism to monitor the quality of messenger RNAs (mRNAs), as exemplified by the translation-dependent recognition and degradation of transcripts harboring premature termination codons (PTCs) by the nonsense mediated mRNA decay (NMD) pathway. We demonstrate in yeast that mRNAs lacking all termination codons are as labile as nonsense transcripts. Decay of "nonstop" transcripts in yeast requires translation but is mechanistically distinguished from NMD and the major mRNA turnover pathway that requires deadenylation, decapping, and 5'-to-3' exonucleolytic decay. These data suggest that nonstop decay is initiated when the ribosome reaches the 3' terminus of the message. We demonstrate multiple physiologic sources of nonstop transcripts and conservation of their accelerated decay in mammalian cells. This process regulates the stability and expression of mRNAs that fail to signal translational termination. PMID- 11910110 TI - Exosome-mediated recognition and degradation of mRNAs lacking a termination codon. AB - One role of messenger RNA (mRNA) degradation is to maintain the fidelity of gene expression by degrading aberrant transcripts. Recent results show that mRNAs without translation termination codons are unstable in eukaryotic cells. We used yeast mutants to demonstrate that these "nonstop" mRNAs are degraded by the exosome in a 3'-to-5' direction. The degradation of nonstop transcripts requires the exosome-associated protein Ski7p. Ski7p is closely related to the translation elongation factor EF1A and the translation termination factor eRF3. This suggests that the recognition of nonstop mRNAs involves the binding of Ski7p to an empty aminoacyl-(RNA-binding) site (A site) on the ribosome, thereby bringing the exosome to a mRNA with a ribosome stalled near the 3' end. This system efficiently degrades mRNAs that are prematurely polyadenylated within the coding region and prevents their expression. PMID- 11910111 TI - Brain to plasma amyloid-beta efflux: a measure of brain amyloid burden in a mouse model of Alzheimer's disease. AB - The deposition of amyloid-beta (Abeta) peptides into amyloid plaques precedes the cognitive dysfunction of Alzheimer's disease (AD) by years. Biomarkers indicative of brain amyloid burden could be useful for identifying individuals at high risk for developing AD. As in AD in humans, baseline plasma Abeta levels in a transgenic mouse model of AD did not correlate with brain amyloid burden. However, after peripheral administration of a monoclonal antibody to Abeta (m266), we observed a rapid increase in plasma Abeta and the magnitude of this increase was highly correlated with amyloid burden in the hippocampus and cortex. This method may be useful for quantifying brain amyloid burden in patients at risk for or those who have been diagnosed with AD. PMID- 11910112 TI - Dynamics of Pleistocene population extinctions in Beringian brown bears. AB - The climatic and environmental changes associated with the last glaciation (90,000 to 10,000 years before the present; 90 to 10 ka B.P.) are an important example of the effects of global climate change on biological diversity. These effects were particularly marked in Beringia (northeastern Siberia, northwestern North America, and the exposed Bering Strait) during the late Pleistocene. To investigate the evolutionary impact of these events, we studied genetic change in the brown bear, Ursus arctos, in eastern Beringia over the past 60,000 years using DNA preserved in permafrost remains. A marked degree of genetic structure is observed in populations throughout this period despite local extinctions, reinvasions, and potential interspecies competition with the short-faced bear, Arctodus simus. The major phylogeographic changes occurred 35 to 21 ka B.P., before the glacial maximum, and little change is observed after this time. Late Pleistocene histories of mammalian taxa may be more complex than those that might be inferred from the fossil record or contemporary DNA sequences alone. PMID- 11910113 TI - Rates of evolution in ancient DNA from Adelie penguins. AB - Well-preserved subfossil bones of Adelie penguins, Pygoscelis adeliae, underlie existing and abandoned nesting colonies in Antarctica. These bones, dating back to more than 7000 years before the present, harbor some of the best-preserved ancient DNA yet discovered. From 96 radiocarbon-aged bones, we report large numbers of mitochondrial haplotypes, some of which appear to be extinct, given the 380 living birds sampled. We demonstrate DNA sequence evolution through time and estimate the rate of evolution of the hypervariable region I using a Markov chain Monte Carlo integration and a least-squares regression analysis. Our calculated rates of evolution are approximately two to seven times higher than previous indirect phylogenetic estimates. PMID- 11910114 TI - A common rule for the scaling of carnivore density. AB - Population density in plants and animals is thought to scale with size as a result of mass-related energy requirements. Variation in resources, however, naturally limits population density and may alter expected scaling patterns. We develop and test a general model for variation within and between species in population density across the order Carnivora. We find that 10,000 kilograms of prey supports about 90 kilograms of a given species of carnivore, irrespective of body mass, and that the ratio of carnivore number to prey biomass scales to the reciprocal of carnivore mass. Using mass-specific equations of prey productivity, we show that carnivore number per unit prey productivity scales to carnivore mass near -0.75, and that the scaling rule can predict population density across more than three orders of magnitude. The relationship provides a basis for identifying declining carnivore species that require conservation measures. PMID- 11910115 TI - Neuronal calcium sensor 1 and activity-dependent facilitation of P/Q-type calcium currents at presynaptic nerve terminals. AB - P/Q-type presynaptic calcium currents (IpCa) undergo activity-dependent facilitation during repetitive activation at the calyx of the Held synapse. We investigated whether neuronal calcium sensor 1 (NCS-1) may underlie this phenomenon. Direct loading of NCS-1 into the nerve terminal mimicked activity dependent IpCa facilitation by accelerating the activation time of IpCa in a Ca2+ dependent manner. A presynaptically loaded carboxyl-terminal peptide of NCS-1 abolished IpCa facilitation. These results suggest that residual Ca2+ activates endogenous NCS-1, thereby facilitating IpCa. Because both P/Q-type Ca2+ channels and NCS-1 are widely expressed in mammalian nerve terminals, NCS-1 may contribute to the activity-dependent synaptic facilitation at many synapses. PMID- 11910116 TI - The medial frontal cortex and the rapid processing of monetary gains and losses. AB - We report the observation of neural processing that occurs within 265 milliseconds after outcome stimuli that inform human participants about gains and losses in a gambling task. A negative-polarity event-related brain potential, probably generated by a medial-frontal region in or near the anterior cingulate cortex, was greater in amplitude when a participant's choice between two alternatives resulted in a loss than when it resulted in a gain. The sensitivity to losses was not simply a reflection of detecting an error; gains did not elicit the medial-frontal activity when the alternative choice would have yielded a greater gain, and losses elicited the activity even when the alternative choice would have yielded a greater loss. Choices made after losses were riskier and were associated with greater loss-related activity than choices made after gains. It follows that medial-frontal computations may contribute to mental states that participate in higher level decisions, including economic choices. PMID- 11910118 TI - Evolution of faster development does not lead to greater fluctuating asymmetry of sternopleural bristle number in Drosophila. AB - Both strong directional selection and faster development are thought to destabilize development, giving rise to greater fluctuating asymmetry (FA), although there is no strong empirical evidence supporting this assertion. We compared FA in sternopleural bristle number in four populations of Drosophila melanogaster successfully selected for faster development from egg to adult, and in four control populations. The fraction of perfectly symmetric individuals was higher in the selected populations, whereas the FA levels did not differ significantly between selected and control populations, clearly indicating that directional selection for faster development has not led to increased FA in sternopleural bristle number in these populations. This may be because: (i) development time and FA are uncorrelated, (ii) faster development does result in FA, but selection has favoured developmentally stable individuals that can develop fast and still be symmetrical, or (iii) the increased fraction of symmetric individuals in the selected populations is an artifact of reduced body size. Although we cannot discriminate among these explanations, our results suggest that the relationship between development time, FA and fitness may be far more subtle than often thought. PMID- 11910117 TI - Control of synaptic strength by glial TNFalpha. AB - Activity-dependent modulation of synaptic efficacy in the brain contributes to neural circuit development and experience-dependent plasticity. Although glia are affected by activity and ensheathe synapses, their influence on synaptic strength has largely been ignored. Here, we show that a protein produced by glia, tumor necrosis factor alpha (TNFalpha), enhances synaptic efficacy by increasing surface expression of AMPA receptors. Preventing the actions of endogenous TNFalpha has the opposite effects. Thus, the continual presence of TNFalpha is required for preservation of synaptic strength at excitatory synapses. Through its effects on AMPA receptor trafficking, TNFalpha may play roles in synaptic plasticity and modulating responses to neural injury. PMID- 11910120 TI - Accelerated molecular evolution of insect orthologues of ERG28/C14orf1: a link with ecdysteroid metabolism? AB - We have analysed the evolution of ERG28/C14orf1, a gene coding for a protein involved in sterol biosynthesis. While primary sequence of the protein is well conserved in all organisms able to synthesize sterols de novo, strong divergence is noticed in insects, which are cholesterol auxotrophs. In spite of this virtual acceleration, our analysis suggests that the insect orthologues are evolving today at rates similar to those of the remaining members of the family. A plausible way to explain this acceleration and subsequent stabilization is that Erg28 plays a role in at least two different pathways. Discontinuation of the cholesterogenesis pathway in insects allowed the protein to evolve as much as the function in the other pathway was not compromised. PMID- 11910121 TI - Genetic variation at minisatellite loci D1S7, D4S139, D5S110 and D17S79 among three population groups of eastern India. AB - Genetic variation at four minisatellite loci D1S7, D4S139, D5S110 and D17S79 in three predominant population groups of eastern India, namely Brahmin, Kayastha and Garo, are reported in this study. The Brahmin and Kayastha are of Indo Caucasoid origin while the Garo community represents the Indo-Mongoloid ethnic group. The methodology employed comprised generation of HaeIII-restricted fragments of isolated DNA, Southern blotting, and hybridization using chemiluminescent probes MS1, pH30, LH1 and V1 for the four loci. All four loci were highly polymorphic in the population groups. Heterozygosity values for the four loci ranged between 0.68 and 0.95. Neither departure from Hardy Weinberg expectations nor evidence of any association across alleles among the selected loci was observed. The gene differentiation value among the loci is moderate (GST = 0.027). A neighbour-joining tree constructed on the basis of the generated data shows very low genetic distance between the Brahmin and Kayastha communities in relation to the Garo. Our results based on genetic distance analysis are consistent with results of earlier studies based on serological markers and linguistic as well as morphological affiliations of these populations and their Indo-Caucasoid and Indo-Mongoloid origin. The minisatellite loci studied here were found to be not only useful in showing significant genetic variation between the populations but also to be suitable for human identity testing among eastern Indian populations. PMID- 11910119 TI - An automated annotation tool for genomic DNA sequences using GeneScan and BLAST. AB - Genomic sequence data are often available well before the annotated sequence is published. We present a method for analysis of genomic DNA to identify coding sequences using the GeneScan algorithm and characterize these resultant sequences by BLAST. The routines are used to develop a system for automated annotation of genome DNA sequences. PMID- 11910122 TI - Isolation and characterization of stable mutants of Streptomyces peucetius defective in daunorubicin biosynthesis. AB - Daunorubicin and its derivative doxorubicin are antitumour anthracycline antibiotics produced by Streptomyces peucetius. In this study we report isolation of stable mutants of S. peucetius blocked in different steps of the daunorubicin biosynthesis pathway. Mutants were screened on the basis of colony colour since producer strains are distinctively coloured on agar plates. Different mutants showed accumulation of aklaviketone, epsilon-rhodomycinone, maggiemycin or 13 dihydrocarminomycin in their culture filtrates. These results indicate that the mutations in these isolates affect steps catalysed by dnrE (mutants SPAK and SPMAG), dnrS (SPFS and SPRHO) and doxA (SPDHC) gene products. PMID- 11910123 TI - Testing quantum dynamics in genetic information processing. AB - Does quantum dynamics play a role in DNA replication? What type of tests would reveal that? Some statistical checks that distinguish classical and quantum dynamics in DNA replication are proposed. PMID- 11910124 TI - Darkness in El Dorado: human genetics on trial. AB - A recent book by a freelance journalist makes major accusations against genetic studies by J. V. Neel in the Amazon a generation ago. Contrary to these charges, there was no connection of Neel's work with human experiments conducted by the Rochester Manhattan project twenty years earlier, nor did the studies serve as a control for survivors of the atomic bombs in Japan. Neel was not a eugenicist. His program of measles vaccination reduced mortality, and was not in any sense an experiment. Given the passage of time and lack of supporting evidence, further investigation of these charges is pointless. However, the political climate in which human populations are studied has changed dramatically over the last generation. Unless guidelines reflect an international consensus, the benefits of population studies to human welfare and science will be jeopardized. The World Health Organization guidelines should be extended to cover current research. PMID- 11910125 TI - Evidence for dominant suppression of repeat-induced point mutation (RIP) in crosses with the wild-isolated Neurospora crassa strains Sugartown and Adiopodoume-7. AB - A convenient assay to score repeat-induced point mutation (RIP) in Neurospora employs the erg-3 locus as a mutagenesis target. Using this assay we screened 132 wild-isolated Neurospora crassa strains for ability to dominantly suppress RIP. RIP was exceptionally inefficient in crosses with the wild isolates Sugartown (P0854) and Adiopodoume-7 (P4305), thereby suggesting the presence of dominant RIP suppressors in these strains. In other experiments, we found no evidence for dominant RIP suppression by the Spore killer haplotypes Sk-2 and Sk-3. PMID- 11910126 TI - K-selection, alpha-selection, effectiveness, and tolerance in competition: density-dependent selection revisited. AB - In the Drosophila literature, selection for faster development and selection for adapting to high density are often confounded, leading, for example, to the expectation that selection for faster development should also lead to higher competitive ability. At the same time, results from experimental studies on evolution at high density do not agree with many of the predictions from classical density-dependent selection theory. We put together a number of theoretical and empirical results from the literature, and some new experimental results on Drosophila populations successfully subjected to selection for faster development, to argue for a broader interpretation of density-dependent selection. We show that incorporating notions of alpha-selection, and the division of competitive ability into effectiveness and tolerance components, into the concept of density-dependent selection yields a formulation that allows for a better understanding of the empirical results. We also use this broader formulation to predict that selection for faster development in Drosophila should, in fact, lead to the correlated evolution of decreased competitive ability, even though it does lead to the evolution of greater efficiency and higher population growth rates at high density when in monotypic culture. PMID- 11910127 TI - Body size and mating success in Drosophila willistoni are uncorrelated under laboratory conditions. AB - Mating activity and wing length were investigated in the F1; progeny of Drosophila willistoni females collected in the field to examine any possible relationship between body size and mating success. The flies were observed in a mating chamber under laboratory conditions. No significant differences in wing length were observed between copulating and noncopulating flies, and there was no significant correlation between wing length and copulation latency for both males and females. These results therefore suggest that the commonly accepted view that large body size is positively correlated with mating success in Drosophila does not always hold true. The results support the view that the extent of environmentally induced variation in body size may be an important factor in determining whether an association between body size and mating success is observed in Drosophila species. PMID- 11910128 TI - Effects of mutations at the stambh A locus of Drosophila melanogaster. AB - We report novel findings on the cytogenetic location, functional complexity and maternal and germline roles of the stambh A locus of Drosophila melanogaster. stmA is localized to polytene bands 44D1.2 on 2R. stmA mutations are of two types: temperature-sensitive (ts) adult and larval paralytic or unconditional embryonic or larval lethal. Twelve alleles reported in this study fall into two intragenic complementing groups suggesting that stmA is a complex locus with more than one functional domain. Some unconditional embryonic lethal alleles show a 'neurogenic' phenotype of cuticle loss accompanied by neural hypertrophy. It is shown that embryos of ts paralytic alleles also show mild neural hypertrophy at permissive temperatures while short exposure to heat induces severe cuticle loss in these embryos. stmA exerts a maternal influence over heat-induced cuticle loss. Unconditional embryonic lethal alleles of stmA are also germline lethal. PMID- 11910131 TI - Necrotizing enterocolitis--an unconquered disease. PMID- 11910129 TI - Male sterility associated with overexpression of the noncoding hsromega gene in cyst cells of testis of Drosophila melanogaster. AB - Of the several noncoding transcripts produced by the hsromega gene of Drosophila melanogaster, the nucleus-limited >10-kb hsromega-n transcript colocalizes with heterogeneous nuclear RNA binding proteins (hnRNPs) to form fine nucleoplasmic omega speckles. Our earlier studies suggested that the noncoding hsromega-n transcripts dynamically regulate the distribution of hnRNPs in active (chromatin bound) and inactive (in omega speckles) compartments. Here we show that a P transposon insertion in this gene's promoter (at -130 bp) in the hsromega05421; enhancer-trap line had no effect on viability or phenotype of males or females, but the insertion-homozygous males were sterile. Testes of hsromega05421; homozygous flies contained nonmotile sperms while their seminal vesicles were empty. RNA:RNA in situ hybridization showed that the somatic cyst cells in testes of the mutant male flies contained significantly higher amounts of hsromega-n transcripts, and unlike the characteristic fine omega speckles in other cell types they displayed large clusters of omega speckles as typically seen after heat shock. Two of the hnRNPs, viz. HRB87F and Hrb57A, which are expressed in cyst cells, also formed large clusters in these cells in parallel with the hsromega-n transcripts. A complete excision of the P transposon insertion restored male fertility as well as the fine-speckled pattern of omega speckles in the cyst cells. The in situ distribution patterns of these two hnRNPs and several other RNA-binding proteins (Hrp40, Hrb57A, S5, Sxl, SRp55 and Rb97D) were not affected by hsromega mutation in any of the meiotic stages in adult testes. The present studies, however, revealed an unexpected presence (in wild-type as well as mutant) of the functional form of Sxl in primary spermatocytes and an unusual distribution of HRB87F along the retracting spindle during anaphase telophase of the first meiotic division. It appears that the P transposon insertion in the promoter region causes a misregulated overexpression of hsromega in cyst cells, which in turn results in excessive sequestration of hnRNPs and formation of large clusters of omega speckles in these cell nuclei. The consequent limiting availability of hnRNPs is likely to trans-dominantly affect processing of other pre-mRNAs in cyst cells. We suggest that a compromise in the activity of cyst cells due to the aberrant hnRNP distribution is responsible for the failure of individualization of sperms in hsromega05421; mutant testes. These results further support a significant role of the noncoding hsromega-n transcripts in basic cellular activities, namely regulation of the availability of hnRNPs in active (chromatin bound) and inactive (in omega speckles) compartments. PMID- 11910130 TI - Mutation analysis of codons 345 and 347 of rhodopsin gene in Indian retinitis pigmentosa patients. AB - More than 100 mutations have been reported till date in the rhodopsin gene in patients with retinitis pigmentosa. The present study was undertaken to detect the reported rhodopsin gene point mutations in Indian retinitis pigmentosa patients. We looked for presence or absence of codon 345 and 347 mutations in exon 5 of the gene using the technique of allele-specific polymerase chain reaction by designing primers for each mutation. We have examined 100 patients from 76 families irrespective of genetic categories. Surprisingly, in our sample the very widely reported highly frequent mutations of codon 347 (P --> S/A/R/Q/L/T) were absent while the codon 345 mutation V --> M was seen in three cases in one family (autosomal dominant form) and in one sporadic case (total two families). This is the first report on codon 345 and 347 mutation in Indian retinitis pigmentosa subjects. PMID- 11910132 TI - XXXIX National Conference of Indian Academy of Pediatrics, Bangalore January 24, 2002. PMID- 11910133 TI - Birth weight patterns in rural undernourished pregnant women. AB - OBJECTIVE: To study the birth weight pattern in chronic as well as currently undernourished pregnant women. DESIGN: Prospective study of rural pregnant women by following eligible women. SETTING: Two adjoining blocks of rural Varanasi. METHOD: 3700 pregnant women from rural areas of Varanasi for whom data for anthropometry, hemoglobin, dietary intake, birth weight, fundal height and abdominal girth at 16 +/- 2, 28 +/- 2 and 36 +/- 2 weeks of gestation were recorded. Outcome measure was birth weight pattern of newborns. RESULTS: Of the births, 7.2% were < 2250 g and 27.4% < 2500 g. The weekly birth weight increments in gestation 36-42 weeks were 5-53 g, only. The fundal height did not increase during 35-39 weeks of gestation (lower by 5 cm as compared to normal). Nutrition supplement in the third trimester significantly increased fundal height and abdominal girth. Fundal height below 24.5 cm at 28 weeks of gestation (1368 women) was associated with higher low birth weight deliveries. CONCLUSION: Birth weight and fundal height increments during later pregnancy are low in undernourished pregnant women. Fundal height < 24.5 cm at 28 weeks of gestation identified women with higher risk for lowbirth weight infants. The prevalence of low birth weight was 27.4% and of prematurity was 6.6%. PMID- 11910134 TI - BCG, tuberculin skin-test results and asthma prevalence in school children in North London. AB - OBJECTIVE: To test whether there is any relationship between asthma prevalence and BCG immunization or tuberculin skin text reaction. DESIGN: Cross-sectional survey. SETTING: Secondary school in Haringey, North London, U.K. SUBJECTS: 780 children aged 11-18 years (median 13.35 years). INTERVENTIONS: Administration of tuberculin skin text and questionnaire. MAIN OUTCOME MEASURES: Diagnosis of asthma, presence of nocturnal cough, exercise-induced wheeze or wheeze with viral respiratory infections; diameter of induration with tuberculin skin text; history of BCG immunization. RESULTS: 57 of 629 children (8.5%) had a significantly positive Mantoux reaction (>or=15 millimeters of induration). Children with and without a history of BCG immunization did not differ significantly in prevalence of asthma diagnosis (11.8% vs 14.1%, p > 0.6), exercise-induced wheeze (16.9% vs 21.2%, p>0.4), viral induced wheeze (15.4% vs 7%, p>0.6) or nocturnal cough (32.3% vs 32.7%, p> 0.6). We also found no significant correlation of the prevalence of asthma diagnosis or symptoms with diameter of Mantoux test reaction. CONCLUSION: There is no evidence of an effect of BCG immunization or tuberculin reactivity on the incidence of asthma in secondary school children in Haringey, North London and the exposure to tuberculosis is high in these children. PMID- 11910135 TI - Atypical pneumonia in children. PMID- 11910136 TI - Retinopathy of prematurity. PMID- 11910137 TI - Role of parents evaluation of developmental status in detecting developmental delay in young children. PMID- 11910138 TI - Hypocalcemia clinical, biochemical, radiological profile and follow-up in a tertiary hospital in India. PMID- 11910139 TI - Transplacentally transmitted anti-measles antibodies in term and preterm infants. PMID- 11910140 TI - Stridor in an infant - a rare cause. PMID- 11910141 TI - Benign familial neonatal convulsions. PMID- 11910143 TI - Incomplete Beckwith-Wiedemann syndrome in a child with orbital rhabdomyosarcoma. PMID- 11910142 TI - Primary hypomagnesemia with secondary hypocalcemia in an infant. PMID- 11910144 TI - An uncommon variety of vein of galen malformation. PMID- 11910145 TI - Congenital neuroblastoma with cutaneous metastases. PMID- 11910146 TI - Staphylococcus warneri septicemia in preterm neonates - a reminder. PMID- 11910147 TI - A countrywide post-marketing surveillance of nimesulide suspension. PMID- 11910148 TI - A case of cutaneous and pharyngeal diphtheria. PMID- 11910149 TI - Pulse oximetry in asphyxiated newborns in the delivery room. PMID- 11910152 TI - Midazolam sedation in mechanically ventilated newborns. PMID- 11910151 TI - Foreign body in the larynx of a one-day-old baby presenting on day 15 of life. PMID- 11910154 TI - Investigation of the structure and phase transitions in the novel A-site substituted distorted perovskite compound Na(0.5)Bi(0.5)TiO(3). AB - Rietveld neutron powder profile analysis of the compound Na(0.5)Bi(0.5)TiO(3) (NBT) is reported over the temperature range 5-873 K. The sequence of phase transitions from the high-temperature prototypic cubic structure (above 813 K), to one of tetragonal (673-773 K) and then rhombohedral structures (5-528 K) has been established. Coexisting tetragonal/cubic (773-813 K) and rhombohedral/tetragonal (with an upper temperature limit of 145 K between 528 and 673 K) phases have also been observed. Refinements have revealed that the rhombohedral phase, space group R3c, with a(H) = 5.4887 (2), c(H) = 13.5048 (8) A, V = 352.33 (3) A(3), Z = 6 and D(x) = 5.99 Mg m(-3), exhibits an antiphase, a( )a(-)a(-) oxygen tilt system, omega = 8.24 (4) degrees, with parallel cation displacements at room temperature. The tetragonal phase, space group P4bm, with a(T) = 5.5179 (2), c(T) = 3.9073 (2) A, V = 118.96 (1) A(3), Z = 2 and D(x) = 5.91 Mg m(-3), possesses an unusual combination of in-phase, a(0)a(0)c(+) oxygen octahedra tilts, omega = 3.06 (2) degrees, and antiparallel cation displacements along the polar axis. General trends of cation displacements and the various deviations of the octahedral network from the prototypic cubic perovskite structure have been established and their systematic behaviour with temperature is reported. An investigation of phase transition behaviour using second harmonic generation (SHG) to establish the centrosymmetric or non-centrosymmetric nature of the various phases is also reported. PMID- 11910153 TI - Twinning and intergrowth of rare earth boride carbides. AB - Twins and intergrown crystals of tetragonal rare earth boride carbides, especially those with the La(5)B(2)C(6) structure type, have been investigated by high-resolution electron microscopy and X-ray diffraction. The structure of the twin interface has been determined. It provides an explanation for coherently intergrown domains of different structure. The Sc(3)C(4) structure type is remarkable because it is frequently intergrown with La(5)B(2)C(6)-type phases. It provides, for instance, a model for the intergrowth of other types, e.g. Gd(4)B(3)C(4) and Gd(5)B(2)C(5). The presence of metal-atom square nets in different orientations in the structures accounts for a number of intergrowth phenomena. The possibilities and limitations of X-ray structure determinations are discussed with respect to actual examples. PMID- 11910155 TI - Intersubsystem chemical bonds in the misfit layer compounds (LaS)(1.13)TaS(2) and (LaS)(1.14)NbS(2). AB - The modulated structures of incommensurate composite crystals (La(0.912)S)(1.13)TaS(2) at room temperature and of (La(0.949)S)(1.14)NbS(2) at T = 115 K are refined against high-resolution X-ray data. The compounds are isostructural with superspace group F'm2m(alpha,0,0)00s. For (LaS)(1.13)TaS(2), lattice parameters of the first subsystem TaS(2) were obtained as a = 3.2922 (1), b = 5.7776 (2) and c = 23.013 (2) A. For the second subsystem LaS, the same b and c parameters were found, but a = 5.8090 (8) A. Refinements led to a final structure model with R = 0.036 for 4767 observed unique reflections (R = 0.023 for 2147 main reflections, R = 0.099 for 1554 first-order satellites and R = 0.112 for 1042 second-order satellites). The final model includes modulation parameters up to the second-order harmonics for the displacements of the atoms, for the occupational parameters and for the temperature parameters. A clear correlation is found between the relative positions of the subsystems, the displacement modulation, the occupational modulation and the modulation of the temperature parameters. The analysis shows that the variations in environments are resolved by correlated variations in the temperature factors. For (LaS)(1.14)NbS(2), lattice parameters at T = 115 K of the NbS(2) subsystem were obtained as a = 3.3065 (4), b = 5.7960 (5) and c = 22.956 (3) A. For the LaS subsystem, the same values for b and c were obtained, but a = 5.7983 (7) A. Refinements led to a final structure model with R = 0.048 for 5909 observed unique reflections (R = 0.034 for 2528 main reflections, R = 0.092 for 2171 first order satellites and R = 0.113 for 1103 second-order satellites). The final structure model is similar to that of (LaS)(1.13)TaS(2). In particular, it is found that the values of the modulation parameters are almost equal and it is concluded that the modulations are independent of the temperature and the replacement of Ta with Nb, and thus represent a general mechanism of resolving the strain between the mutually incommensurate layers. PMID- 11910156 TI - Single-crystal study of the m = 2 tubular cobalt oxide, Bi(4)Sr(12)Co(8)O(30 delta). AB - The structure of the m = 2 tubular compound Bi(4)Sr(12)Co(8)O(30-delta), bismuth strontium cobalt oxide, was determined by single-crystal X-ray diffraction. This phase of orthorhombic symmetry exhibits a very strong tetragonal pseudosymmetry. The structure consists of 90 degrees -oriented Bi(2)Sr(2)CoO(6+delta) slices, four Co atoms wide, forming [Sr(4)Co(4)O(13)](infinity) pillars at their intersection. The Co atoms in these pillars form four corner-sharing CoO(5) bipyramids. In the resulting [Co(4)O(13)] cluster, an anionic disorder is evidenced and discussed. Then, an accurate description of the particular structure of the pillars is given. Finally, a comparison with the Mn tubular compound Bi(3.6)Sr(12.4)Mn(8)O(30-delta) is carried out. PMID- 11910157 TI - Crystal chemistry of zirconosilicates and their analogs: topological classification of MT frameworks and suprapolyhedral invariants. AB - The first attempt is undertaken to consider systematically topological structures of zirconosilicates and their analogs (60 minerals and 34 synthetic phases), where the simplest structure units are MO(6) octahedra and TO(4) tetrahedra united by vertices ([TO(4)]:[MO(6)] = 1:1-6:1). A method of analysis and classification of mixed three-dimensional MT frameworks by topological types with coordination sequences [N(k)] is developed, which is based on the representation of crystal structure as a finite "reduced" graph. The method is optimized for the frameworks of any composition and complexity and implemented within the TOPOS3.2 program package. A procedure of hierarchical analysis of MT-framework structure organization is proposed, which is based on the concept of polyhedral microensemble (PME) being a geometrical interpretation of coordination sequences of M and T nodes. All 12 theoretically possible PMEs of MT(6) polyhedral composition are considered where T is a separate and/or connected tetrahedron. Using this methodology the MT frameworks in crystal structures of zirconosilicates and their analogs were analyzed within the first 12 coordination spheres of M and T nodes and related to 41 topological types. The structural correlations were revealed between rosenbuschite, lavenite, hiortdahlite, woehlerite, siedozerite and the minerals of the eudialyte family. PMID- 11910158 TI - A new set of molecular descriptors. AB - A new set of criteria for quantitative analysis of molecular interactions is proposed, which is based on the conceptions of atomic and molecular Voronoi Dirichlet polyhedra. It is shown that the calculation of solid angles of ligands and complexes as a whole allows one to estimate screening effects and the probability of forming intra- and intercomplex non-valent contacts. The set proposed was used to study the influence of steric factors on the stability of complex groups as well as on the presence or absence of agostic contacts in crystal structures of 808 rare-earth pi-complexes. PMID- 11910159 TI - Modified photoreactivity due to mixed crystal formation. I. Three mixed crystals between isostructural cobaloxime complexes. AB - Three crystals of 2-cyanoethyl cobaloxime complexes with 3-chloropyridine, 3 bromopyridine and 3-methylpyridine as axial base ligands are isostructural to one another. Three mixed crystals were formed between the pairs: (3-bromopyridine)(2 cyanoethyl)cobaloxime-(2-cyanoethyl)(3-methylpyridine)cobaloxime(0.45/0.55);(3 chloropyridine)(2-cyanoethyl)cobaloxime-(2-cyanoethyl)(3bromopyridine)cobaloxime (0.61/0.39); (3-chloropyridine)(2-cyanoethyl)cobaloxime-(2-cyanoethyl)(3 methylpyridine)cobaloxime (0.44/0.56). The X-ray crystal structure analysis revealed that the mixed compounds are also isostructural to the component crystals. It was found from the change in IR spectra that the 2-cyanoethyl groups in the three mixed crystals isomerized to 1-cyanoethyl groups on exposure to a xenon lamp, as observed for the 2-cyanoethyl groups in the component crystals. Rate constants for the three mixed and three component crystals, measured under the same conditions, are approximately the same, as the reaction cavities for the 2-cyanoethyl groups in the six crystals have almost the same size. For the mixed crystal between 3-chloropyridine and 3-methylpyridine complexes, the isomerization proceeded with retention of the single-crystal form. The conformation and configuration of the 1-cyanoethyl group that was produced were well explained by the shape of the reaction cavity before irradiation. PMID- 11910160 TI - Structure prediction as a tool for solution of the crystal structures of metallo organic complexes using powder X-ray diffraction data. AB - A simulated-annealing direct-space approach has been applied to predict the crystal structures of a series of metallo-organic complexes containing Zn, Cu and Ni. The prediction methodology generates a set of energetically reasonable crystal structures among which the actual structure is present, but it is not always possible to specify unambiguously the known crystal structure solely on the basis of energy. In each case, however, the ambiguity may be resolved by recourse to laboratory powder X-ray diffraction (PXRD) data. In this manner, structure prediction is shown to be a powerful tool for structure solution using PXRD data, with the additional advantage that indexing of the PXRD profile is not required at the outset. PMID- 11910161 TI - Structures of trans-[PtCl(2)(PBz(3))(2)], trans-[PtI(2)(PBz(3))(2)], trans [Pt(NCS)(2)(PBz(3))(2)].0.5C(6)H(6) and trans-[PdI(2)(PBz(3))(2)]. AB - A series of structures of trans-[MX(2)(PBz(3))(2)] [M = Pt, X = Cl(-); PBz(3) = tribenzylphosphine (1), I(-), trans-diiodobis(tribenzylphosphine)platinum(II) (2), and NCS(-), trans-di(thiocyanate)bis(tribenzylphosphine)platinum(II) (3); M = Pd, X = I(-), trans-diiodobis(tribenzylphosphine)palladium(II) (4)] have been characterized by X-ray crystallography. In all compounds each tribenzylphosphine has one benzylcarbon close to the coordination plane. In (1), (2) and (4) those (in-plane) C atoms, from the two different PBz(3), exhibit an anti conformation along the P-P axis, while (3) has the gauche conformation. Root mean square (RMS) calculations and half-normal probability plots show that the complexes in (2) and (4) are very similar and the only significant differences between them are the M P bonds, 2.354 (4) and 2.330 (5) A, and the M-I bond distances, 2.604 (1) and 2.611 (2) A, for Pd and Pt, respectively. Calculations of the steric demand of the PBz(3) ligands based on the Tolman model gave values ranging from 155 to 178 degrees for the effective and 156 to 179 degrees for the Tolman angles, respectively. PMID- 11910162 TI - Inclusion compounds of a diol host with xylidines: controlled stoichiometries. AB - The diol host, 1,1'-bis-(4-hydroxyphenyl)cyclohexane (DHPC) and a number of xylidine isomers as guests formed a series of inclusion compounds that gave rise to various host:guest ratios controlled by crystallization temperatures. For the host DHPC with a particular xylidine isomer, the number of guests included generally decreases as the crystallization temperature increases. The crystal structures of these host-guest compounds were elucidated using single-crystal X ray diffraction. Their thermal stabilities were characterized by TG and DSC analysis. The selectivity of enclathration by the host was measured by carrying out a series of competition experiments. The kinetics of guest decomposition were studied using isothermal and non-isothermal methods and reconciled with the crystal structures. PMID- 11910163 TI - Structure and pseudosymmetry of cholesterol at 310 K. AB - The structure of cholesterol above the (304.8 K) phase transition, previously published in preliminary form [Hsu & Nordman (1983). Science, 220, 604-606], has been fully refined using augmented X-ray data. The crystals are triclinic, space group P1, with (reassigned) cell parameters a = 27.565 (10), b = 38.624 (16), c = 10.748 (4) A, alpha = 93.49 (3), beta = 90.90 (3), gamma = 117.15 (3) degrees, and V = 10151 (7) A(3). The unit cell contains Z = 16 molecules, of which eight are related to the other eight by unusual twofold rotational pseudosymmetry. The structure is related to the room-temperature phase, with Z = 8, by a rearrangement of some of the molecules, and by a doubling of the a axis. PMID- 11910164 TI - Isostructuralism in a series of methylester/methylamide derivatives of (R,R)-O,O' dibenzoyl tartaric acid; inclusion properties and guest-dependent homeotypism of the crystals of (R,R)-O,O'-dibenzoyltartaric acid diamide. AB - The compounds studied are methyl ester, amide and methylamide derivatives of (R,R)-O,O'-dibenzoyl tartaric acid. The molecules adopt the planar T conformation of the four-carbon chain with the terminal C=O bonds situated antiperiplanar with respect to the nearest C*-O bond. All investigated molecules occupy a twofold symmetry site in the crystal, including the mono(N-methylamide) monomethylester which lacks the C(2) molecular symmetry. Connected with this is the static disorder in which the methylester and the N-methylamide groups replace each other and isostructuralism within the methylester/methylamide series. (R,R)-O,O' Dibenzoyltartaric acid diamides [(R,R)-O,O'-dibenzoyl-2,3 dihydroxybutanediamides], both primary and secondary, form hydrogen-bond aggregation patterns typical for amides, despite the presence of other hydrogen bond acceptors in the molecule. However, in primary amides such packing leads to the creation of homeotypic crystal structures in which structural voids are filled by cyclic solvent molecules (pyridine, 1,4-dioxane). The presence of polyamide ladders, consisting of 'fused' hydrogen-bond rings, seems to be responsible for the low solubility and high melting point of these substances. PMID- 11910165 TI - Chiral versus racemic building blocks in supramolecular chemistry: tartrate salts of organic diamines. AB - In the 1:1 adducts C(12)H(10)N(2).C(4)H(6)O(6) formed between 1,2-bis(4' pyridyl)ethene and racemic tartaric acid [(I), triclinic P1;, Z' = 1] and (2R,3R) tartaric acid [(II), triclinic P1, Z' = 2], the ionic components are linked by hard hydrogen bonds into single sheets, which are further linked by C-H.O hydrogen bonds. In the analogous adducts C(10)H(18)N(2).C(4)H(6)O(6) formed by 4,4'-bipyridyl with racemic tartaric acid [(III), triclinic P1;, Z' = 1] and the chiral acid [(IV), monoclinic P2(1), Z' = 1], the hard hydrogen bonds generate bilayers which are again linked by C-H.O hydrogen bonds. Piperazine forms a 1:1 salt [((C(4)H(10)N(2))H(2))(2+)].[(C(4)H(4)O(6))(2-)] with (2R,3R)-tartaric acid [(V), monoclinic P2(1)] sheets, which are linked by the cations to form a pillared-layer framework. In each of the 1:2 salts formed by racemic tartaric acid with piperazine [(VI), monoclinic P2(1)/n, Z' = 0.5] and 1,4 diazabicyclo[2.2.2]octane (DABCO) [(VII), monoclinic P2(1)/n, Z' = 0.5], the cation lies across a centre of inversion, with the [(HN(CH(2)CH(2))(3)NH)(2+)] cation disordered over two sets of sites: in both (VI) and (VII) the anions form a three-dimensional framework encapsulating large voids which accommodate the cations. The salt formed between DABCO and (2R,3R)-tartaric acid [(VIII), orthorhombic P2(1)2(1)2(1), Z' = 1] has 3:4 stoichiometry and contains four different types of ion, [(HN(CH(2)CH(2))(3)NH)(2+)](2).[N(CH(2)CH(2))(3)NH](+).3(C(4)H(5)O(6))( ).C(4)H(4)O(6)(2-): the hard hydrogen bonds generate a three-dimensional framework. PMID- 11910166 TI - Polymorphs and pseudopolymorphs of N,N'-dithiobisphthalimide. AB - N,N'-Dithiobisphthalimide, C(16)H(8)N(2)O(4)S(2) (I), forms a wide range of polymorphs and solvates (pseudopolymorphs). When (I) is crystallized from methanol it yields a solvent-free polymorph (4), in Pna2(1) with Z' = 1, in which the molecules are linked into chains by a single C-H.O hydrogen bond: crystallization from either acetonitrile or dimethylformamide produces a monoclinic polymorph (5), in P2(1)/c with Z' = 2, also solvent-free, in which the molecules are linked into molecular ladders. Nitromethane forms a monosolvate, C(16)H(8)N(2)O(4)S(2).CH(3)NO(2) (6), in P2(1)/c with Z' = 1, in which the solvent molecules are linked to the molecules of (I) not only via a conventional C-H.O hydrogen bond but also via a polarized multicentre interaction involving all three C-H bonds of the solvent molecule. Chlorobenzene forms a precise hemisolvate, C(16)H(8)N(2)O(4)S(2).0.5C(6)H(5)Cl (7), in P1; with Z' = 1, while ethylbenzene forms an approximate hemisolvate 2C(16)H(8)N(2)O(4)S(2).0.913C(6)H(5)C(2)H(5).0.087H(2)O (8), in P2(1)/c with eight molecules of (I) per unit cell. In both solvates the molecules of (I) are linked, in (7) by pi.pi stacking interactions augmented by weak C-H.O hydrogen bonds and in (8) by stronger C-H.O hydrogen bonds: the solvent molecules lie in isolated cavities, disordered across inversion centres in (7) and fully ordered in general positions in (8). Crystallization of (I) either from tetrahydrofuran or from wet tert-butanol yields isomorphous solvates (9) and (10), respectively, in C2/c with Z' = 0.5, in which molecules of (I) lie across twofold rotation axes and are linked by pi.pi stacking interactions and very weak C-H.O hydrogen bonds, forming a framework enclosing continuous channels: highly disordered solvent molecules lie within these channels. p-Xylene and toluene form isomorphous hemisolvates (11) and (12) with unit cells metrically very similar to those of (9) and (10), but in P2(1)/n with Z' = 1: in these two solvates the molecules of (I) are linked into a framework by very short C-H.O hydrogen bonds; the solvent molecules lie within continuous channels, but they are localized across inversion centres so that the toluene is disordered across an inversion centre. PMID- 11910167 TI - Amino-substituted O(6)-benzyl-5-nitrosopyrimidines: interplay of molecular, molecular-electronic and supramolecular structures. AB - The structures of eight 2,4,6-trisubstituted-5-nitrosopyrimidines (one of which crystallizes in two polymorphs) have been determined, including seven O(6)-benzyl derivatives which are potential, or proven, in vitro inhibitors of the human DNA repair protein O(6)-alkylguanine-DNA-transferase. In the derivatives having an amino substituent at the 4-position, an intramolecular N-H.O hydrogen bond with the nitroso O as an acceptor leads to an overall molecular shape similar to that of substituted purines. There is a marked propensity for these nitroso compounds to crystallize with Z' = 2. The structure of an analogue with no nitroso group is also reported for comparative purposes. Compounds containing the N-alkyl substituents -NHCH(2)COOEt, -NHCH(2)CH(2)COOEt and -NHCH(CH(2)Ph)COOEt, derived from amino acid esters, exhibit a rich variety of conformational behaviour, and in all of the nitroso compounds the bond lengths provide strong evidence for a highly polarized electronic structure. Associated with this polarization is extensive charge-assisted hydrogen bonding between the molecules, leading to supramolecular aggregation in the form of finite (zero-dimensional) aggregates, chains, molecular ladders, sheets and frameworks. PMID- 11910170 TI - NADPH-diaphorase activity and nitric oxide synthase-like immunoreactivity colocalize in the electromotor system of four species of gymnotiform fish. AB - The electric organ discharge (EOD) of gymnotiform electric fish is controlled by a well-characterized neural circuit in the brainstem and spinal cord. NADPH diaphorase (NADPH-d) activity was previously found in phase-locking and/or rapidly firing neurons in the electromotor and electrosensory systems of Apteronotus leptorhynchus [Turner and Moroz, 1995]. These findings suggested that nitric oxide synthase (NOS) is expressed in these neurons and may regulate their precise, high frequency firing. We extended these results by examining the distribution of both NADPH-d activity and NOS-like immunoreactivity (NOS-lir) in the electromotor systems of four gymnotiform species that differ in the frequency and modulation of their EODs. NOS-lir colocalized with NADPH-d staining throughout the electromotor system, indicating that NADPH-d is a faithful indicator of NOS in this system. The distribution of NOS-lir and NADPH-d was similar in the electromotor systems of all four species in this study, with one exception: NOS and NADPH-d staining was consistently less intense in pacemaker and relay cells in Sternopygus macrurus, which produces low frequency EODs, than in the three other species that produce higher frequency EODs. This species difference in NOS expression in the pacemaker nucleus may be related to species differences either in EOD frequency or in modulations of the EOD (e.g., the jamming avoidance response). In Apteronotus species, NOS-lir and NADPH-d were concentrated in bands along the axons of their nerve-derived electric organs. These bands corresponded to regions surrounded by little or no staining with a Schwann cell-specific antibody, suggesting that the NOS-positive regions lie near nodes of Ranvier. In Sternopygus and Eigenmannia, the innervated, posterior membranes of muscle-derived electrocytes were more intensely labeled for NADPH-d and NOS than inexcitable portions of the membrane. Thus, in both muscle- and nerve-derived electric organs, NOS is concentrated near excitable membranes. These results indicate that NOS is well-positioned within the electromotor system to regulate the frequency, precision, amplitude, and waveform of EODs. PMID- 11910172 TI - Sciaenid inner ears: a study in diversity. AB - Sciaenid fishes (Family Sciaenidae) could potentially serve as models for understanding the relationship between structure and function in the teleost auditory system, as they show a broad range of variation in not only the structure of the ear but also in the relationship between the ear and swim bladder. In this study, scanning electron microscopy (SEM) was used to investigate inner ear ultrastructure of the Atlantic croaker (Micropogonias undulatus), spotted seatrout (Cynoscion nebulosus), kingfish (Menticirrhus americanus) and spot (Leiostomus xanthurus). These species reflect the diversity of otolith and swim bladder morphology in sciaenids. The distribution of different hair cell bundle types, as well as hair cell orientation patterns on the saccular and lagenar maculae of these fishes were similar to one another. The rostral ends of the saccular sensory epithelia (maculae) were highly expanded in a dorsal-ventral direction in the Atlantic croaker and spotted seatrout as compared to the kingfish and spot. Also, ciliary bundles of the saccular maculae contained more stereocilia in the Atlantic croaker and spotted seatrout as compared with kingfish and spot. The shapes of the lagenar maculae were similar in all four species. In the Atlantic croaker and spotted seatrout lagenar maculae, the number of stereocilia per bundle was greater than those for the kingfish and spot. Given that saccular macula shape and numbers of stereocilia per bundle correlate with swim bladder proximity to the ear in the studied species, it is possible that inner ear ultrastructure could be indicative of auditory ability in fishes. PMID- 11910171 TI - Evolution of calls and auditory tuning in the Physalaemus pustulosus species group. AB - In species within the Physalaemus pustulosus species group, male frogs produce a whine-like advertisement call consisting of a frequency sweep typically descending from 1,000 to 400 Hz (depending on the species). One species, Physalaemus pustulosus, the tungara frog, has evolved a second call syllable, the chuck, which males place after their whine. Most energy in the chuck is above 1,500 Hz and peaks at 2,400 Hz. We investigated whether the evolution of this new call component in P. pustulosus coincided with evolution of auditory tuning. We used multiunit electrophysiological recordings of auditory-evoked activity in the midbrain to characterize auditory tuning in Physalaemus pustulosus, four other Physalaemus species within the P. pustulosus clade, and three additional, closely related Physalaemus species as outgroups. All eight species had similar sensitivity profiles, with a broad area of enhanced sensitivity from 100 to 1,100 Hz, which we presume represents amphibian papilla (AP) tuning, and a second, narrower area of enhanced sensitivity centered above 2,100 Hz, which we presume represents basilar papilla (BP) tuning. For all species, the whine stimulates the AP. The P. pustulosus chuck stimulates the BP. The frequency with greatest AP sensitivity differed significantly among species. Although in all cases the AP peak lay within the frequency sweep of the whine, phylogenetically corrected correlations revealed no significant relationships between AP tuning and any spectral feature of the whine. BP tuning was similar among all species, with mean BP best excitatory frequencies (BEFs) around 2,100-2,200 Hz, with the exception of P. pustulatus, with a mean BP BEF of 2,549 Hz. Physalaemus pustulosus, the only investigated species that produces a call component stimulating the BP, had a BP BEF that was not significantly different from any of the species within its clade except P. pustulatus, or from any of the outgroup species. A phylogenetic reconstruction of ancestral BP tuning confirms that the only point of evolutionary change in BP tuning is in the line of descent leading to P. pustulatus, not in the line leading to P. pustulosus despite this being the species using the BP for communication. The results indicate that BP tuning around 2,200 Hz is a conserved trait in the Physalaemus pustulosus species group and that no evolution of BP tuning accompanied the subsequent evolution of the call component (the chuck) that stimulates it. This supports the sensory exploitation idea, which posits that signals evolve to match preexisting features of receiver systems. PMID- 11910173 TI - The role of temporal cues in rhesus monkey vocal recognition: orienting asymmetries to reversed calls. AB - An understanding of the acoustic cues that animals use to categorize their vocalizations has important implications for the way we design neuroethological investigations of auditory function. Compared to other species, we know relatively little about the kinds of acoustic features used by nonhuman primates to recognize and categorize vocalizations. To further our understanding, this study explores the role of temporal features in recognition of conspecific vocalizations by rhesus macaques (Macaca mulatta). Experiments were designed to extend an earlier set of findings showing that adult rhesus macaques selectively turn with the right ear leading when a conspecific vocalization is played 180 degrees behind them, but turn left or not at all when a non-conspecific signal is played. Two call types were used as stimuli: shrill barks (alarm call) and harmonic arches (food call). We found that for normal calls, rhesus macaques turned to the right - supporting earlier findings - but for time- reversed shrill barks and harmonic arches, subjects oriented to the left. These results suggest that for at least a subset of calls, rhesus macaques use temporal cues to recognize conspecific vocal signals. The asymmetry of the behavioral response, and the corresponding asymmetry in the time-amplitude waveform, may have important implications for studies of temporal coding in the primate auditory system. PMID- 11910174 TI - Brain structures of a medaka mutant, el (eyeless), in which eye vesicles do not evaginate. AB - Eye development and brain structures of a mutant teleost fish were investigated. The el (eyeless) mutation in medaka (Oryzias latipes) is recessive and affects eye formation; in the most severe cases, it results in the absence of eyes. Developmental studies revealed that normal eyeballs are not formed in the el mutant embryos, but small optic cup-like structures differentiate in situ in the walls of the prosencephalon without evagination. The anophthalmic el homozygous fish hatched normally, although they did not respond behaviorally to visual stimuli. A small fraction of these fish grew to adulthood. In the adult anophthalmic el homozygous fish, the brain exhibited abnormalities in several subdivisions. A pair of small abnormal protrusions was observed on the surface of the ventral telencephalon and preoptic area. Immunocytochemistry using a rhodopsin monoclonal antibody showed that opsin-positive cells were present in the abnormal structures. Bodian staining showed that the optic nerves were present near the abnormal structures, although the number of optic nerve fibers was extremely small. The optic tectum was extremely small, and the thickness of the stratum opticum and stratum fibrosum et griseum superficiale was reduced. These behavioral and morphological observations suggest that the adult anophthalmic el homozygous fish are functionally blind, although small retina like structures were partially differentiated and persisted in the adult fish brain. Moreover, the adult anophthalmic el homozygous fish were infertile, and the sizes of the hypophysis and the hypothalamus were reduced. Thus, the el mutation affects not only the brain structures that are related to the visual system but also those related to the reproductive system. PMID- 11910175 TI - International Conference on Exogenous Factors Affecting Thrombosis and Haemostasis. Paris, France, July 12-13, 2001. Proceedings. PMID- 11910176 TI - Vampire bat plasminogen activator DSPA-alpha-1 (desmoteplase): a thrombolytic drug optimized by natural selection. AB - Plasminogen activators are enzymes found in all vertebrate species investigated so far. Their physiological function is the generation of localized proteolysis in the context of tissue remodeling, wound healing and neuronal plasticity. The common vampire bat (Desmodus rotundus) is a New World species that feeds exclusively on blood. Its saliva contains highly potent plasminogen activators, specialized in rapid lysis of fresh blood clots. Biochemical and pharmacological evidence indicates that these plasminogen activators represent a new class of thrombolytics with pharmacological and toxicological properties superior to human tissue-type plasminogen activator, the clot dissolving agent now most frequently used in medicine. A form of the enzyme produced by recombinant DNA technology is currently employed to test this hypothesis in clinical studies. PMID- 11910177 TI - Proteases from Vipera lebetina venom affecting coagulation and fibrinolysis. AB - Our studies of the venom from the Levantine viper Vipera lebetina have demonstrated the existence of both coagulants and anticoagulants in the same venom. We showed that V. lebetina venom contains: (1) proteases that degrade fibrinogen, but not fibrin; (2) fibrinolytic enzyme (lebetase); (3) factor X activator (VLFXA); (4) factor V activator (VLFVA). Fibrinolytic enzyme and VLFXA are metalloproteases; the other studied enzymes are serine proteases. alpha Fibrinogenase has no homolog among known serine proteases. Beta-fibrinogenase is a typical thermostable arginine esterase that hydrolyzes esters and amides of arginine and attacks the beta-chain of fibrinogen. Lebetase is a direct-acting fibrinolytic zinc metalloendopeptidase related in amino acid sequence to reprolysins. We used the matrix-assisted laser desorption/ionization time-of flight mass spectrometry technique for the recovery and identification of peptides released by protease hydrolysis and for the detection of human factor X cleavage products after VLFXA hydrolysis. VLFXA cleaves the Arg(52)-Ile(53 )bond in the heavy chain of human factor X and the Arg(226)-Val(227) bond in human factor IX precursor; VLFVA cleaves Arg(1545)-Ser(1546) in factor V. PMID- 11910178 TI - Snake venom proteinases as tools in hemostasis studies: structure-function relationship of a plasminogen activator purified from Trimeresurus stejnegeri venom. AB - Snake venom serine proteinases affect many steps of the blood coagulation cascade. Each of them usually acts selectively on one coagulation factor. They are therefore potentially useful components to study the mechanisms of action, the regulation and the structure-function relationships of human serine proteinase coagulation factors. This strategy is illustrated for a plasminogen activator purified from Trimeresurus stejnegeri venom. PMID- 11910179 TI - Alfimeprase: pharmacology of a novel fibrinolytic metalloproteinase for thrombolysis. AB - Alfimeprase is a recombinantly produced, truncated form of fibrolase, a known directly fibrinolytic zinc metalloproteinase that was first isolated from the venom of the southern copperhead snake (Agkistrodon contortrix contortrix). Both fibrolase and alfimeprase have been shown to have direct proteolytic activity against the fibrinogen Aalpha chain. In vivo pharmacology studies have shown that thrombolysis with alfimeprase is up to 6 times more rapid than with plasminogen activators. Alfimeprase can be bound and neutralized by serum alpha(2) macroglobulin, a prevalent mammalian protease inhibitor which is capable of forming a macromolecular complex with alfimeprase. As a result, systemic bleeding complications have been greatly reduced due to the inhibitory effects of alpha(2) macroglobulin. This article reviews the biochemical in vitro and in vivo characteristics of this novel acting thrombolytic. PMID- 11910180 TI - Multifunctional snake C-type lectins affecting platelets. AB - Snake venoms contain a wide range of components, many of which affect haemostasis by activation or inhibition of platelets or coagulation factors. They can be classified into groups based on structure and mode of action. One group is the snake C-type lectins, so called because of the typical folding which closely resembles that found in classical C-type lectins, such as selectins and mannose binding proteins. Unlike the classic C-type lectins, those from snakes are generally heterodimeric with two subunits, alpha and beta. Some are multimeric heterodimers. The subunits have homologous sequences and are generally linked by a disulphide bond as well as by swapping loops. One of the first C-type lectins with a defined function was echicetin which was demonstrated to bind to platelet GPIb and block several functions of this receptor. Since then, many proteins with similar structure have been reported to act on platelet receptors or coagulation factors and several have been crystallized. These proteins were thought to be specific for a single platelet receptor or coagulation factor, i.e. they had only one receptor per heterodimer. Recent studies show that most of these C-type lectins have binding sites for more than one ligand and have complex mechanisms of action. PMID- 11910181 TI - The use of snake venom toxins as tools to study platelet receptors for collagen and von Willebrand factor. AB - A large proportion of the biologically active proteins and peptides present within snake venoms interact with components of the haemostatic system to promote or inhibit the normal sequence of events that lead to clot formation. The venom proteins achieve their effects through interaction with various components of the coagulation cascade, endothelial matrix and platelets. Within the latter group, a number of venom proteins target the interaction of platelets with the major adhesive proteins, von Willebrand factor and collagen. The venom proteins bind either the adhesive protein itself or their receptors on the platelet surface, notably GP-Ib-IX-V and GPVI. This review discusses the substantial contribution that venom proteins have made to our understanding of the role of these two adhesive proteins and their receptors (excluding GPIIb-IIIa) in platelet regulation. PMID- 11910182 TI - Lebecetin, a C-lectin protein from the venom of Macrovipera lebetina that inhibits platelet aggregation and adhesion of cancerous cells. AB - A novel C-lectin protein, lebecetin, was purified and characterized from the venom of Macrovipera lebetina. It is a disulfide-linked heterodimer of 15 and 16 kD. The subunits are homologous to each other and to the other snake venom proteins of the C-type (Ca(2+)-dependent) lectin superfamily. Lebecetin shows a potent inhibitory effect on whole blood and washed platelets induced by different agonists. It inhibits the agglutination of human fixed platelets in the presence of ristocetin. Lebecetin also interferes with the adhesion of IGR39 melanoma and HT29D4 adenocarcinoma cells. These two lines adhere to lebecetin used as matrix. Lebecetin is also able to strongly reduce IGR39 and HT29D4 cell adhesion to fibrinogen and laminin, but not to fibronectin and collagen types I and IV, respectively. Adhesion properties of lebecetin may thus involve integrin receptors. PMID- 11910183 TI - New insights on disintegrin-receptor interactions: eristostatin and melanoma cells. AB - Viper venom disintegrins have been used frequently to study the cellular receptors which characterize various types of cells, including platelets, endothelial cells and cancer cells. While the majority of such analyses have pointed to involvement of integrin receptors alphavbeta3, alpha5beta1 or alphaIIbbeta3, this may not always be so. Eristostatin, from Eristocophis macmahoni, is a potent inhibitor of ADP-induced platelet aggregation as well as of human and murine melanoma metastases in mouse model systems. This disintegrin requires an RGDW motif, as well as an intact C-terminus, in order to interact with both platelets and four different types of melanoma cells. Eristostatin causes nonmetastatic SBc12 melanoma cells to show higher susceptibility to specific killing by NK-like TALL-104 cells. While it is known that eristostatin binds to alphaIIbbeta3 on platelets, the receptor with which eristostatin binds to the melanoma cells has not yet been identified. PMID- 11910184 TI - A novel snake venom disintegrin that inhibits human ovarian cancer dissemination and angiogenesis in an orthotopic nude mouse model. AB - OVCAR-5 is a human epithelial carcinoma cell line of the ovary, established from the ascitic fluid of a patient with progressive ovarian adenocarcinoma without prior cytotoxic treatment. The unique growth pattern of ovarian carcinoma makes it an ideal model for examining the anticancer activity of contortrostatin (CN), a homodimeric disintegrin from southern copperhead venom. FACS analysis revealed that OVCAR-5 is integrin alphavbeta3 negative, but alphavbeta5 positive. CN effectively blocks the adhesion of OVCAR-5 cells to several extracellular matrix proteins and inhibits tumor cell invasion through an artificial basement membrane. In a xenograft nude mouse model with intraperitoneal introduction of OVCAR-5 cells, intraperitoneal injection of CN was used for therapy. Tumor dissemination in CN-treated versus control groups was studied by gross examination, and antiangiogenic potential was examined by factor VIII immunohistochemistry and image analysis. CN not only significantly inhibited ovarian cancer dissemination in the nude mouse model, but it also dramatically prevented the recruitment of blood vessels to tumors at secondary sites. PMID- 11910185 TI - Viper venom components affecting angiogenesis. AB - Angiogenesis is a complex process consisting of the proliferation, migration and differentiation of endothelial cells, and it is essential for the progression of malignant solid tumors. In this report, we examine the effects of disintegrins (e.g. rhodostomin and accutin) and glycoprotein-lb-binding proteins (e.g. agkistin) on each step in angiogenesis using in vitro and in vivo models. Rhodostomin (but not agkistin) inhibited the viability of human umbilical vein endothelial cells (HUVECs) and capillary tube formation of HUVECs. Rhodostomin also inhibited HUVEC migration and invasion evoked by basic fibroblast growth factor (bFGF). In in vivo studies, rhodostomin inhibited bFGF-, but not vascular endothelial-growth-factor (VEGF)- associated angiogenesis in chick chorioallantoic membrane model, blocked both bFGF and B16F10 melanoma cell induced neovascularization, and suppressed the growth of subcutaneously inoculated B16F10 solid tumor, leading to a prolonged survival of the C57BL/6 mice treated with rhodostomin. The antiangiogenic effects of rhodostomin on bFGF treated HUVECs may be mainly related to the blockade of the interaction of endothelial alpha(v)beta(3) and extracellular matrix. PMID- 11910186 TI - Lebetin peptides: potent platelet aggregation inhibitors. AB - Lebetins from Macrovipera lebetina snake venom constitute a new class of inhibitors of platelet aggregation. There are two groups of peptides: lebetin 1 (L1; 11- to 13-mer) and lebetin 2 (L2; 37- to 38-mer). The short lebetins are identical to the N-terminal segments of the longer ones. They inhibit platelet aggregation induced by various agonists (e.g. thrombin, PAF-acether or collagen). The shortest lebetin (11-mer) shows potent inhibition of rabbit (IC(50) = 7 nM) and human (IC(50) = 5 nM) platelets. They prevent collagen-induced thrombocytopenia in rats. N- and C-terminal-truncated synthetic L1gamma (sL1gamma; 11-mer) is less active in inhibiting platelet aggregation than the native peptide. Results from Ala scan studies of the sL1gamma peptide indicated that replacement of the residues (P3, G7, P8, P9 or N10) resulted in a remarkable drop in the activity, whereas replacement of residues K2, P4 or K6 by Ala resulted in enhancement of the antiplatelet activity by at least 10-fold. To examine the activity of multimeric L1gamma, several multimeric peptides were synthesized using the multiple-antigen peptide system assembled on a branched lysine core and their antiplatelet activity was evaluated in vitro. The largest multimeric peptides showed a 1,000-fold increase in antiplatelet activity. PMID- 11910187 TI - Diagnostic uses of snake venom. AB - Snake venom toxins are invaluable for the assay of coagulation factors and for the study of haemostasis generally. Thrombin-like enzymes (SVTLE) are used for fibrinogen and fibrinogen breakdown product assays as well as detecting dysfibrinogenaemias. Since SVTLE are not inhibited by heparin, they can be used for assaying antithrombin III in samples containing heparin. Snake venom prothrombin activators are utilised in prothrombin assays, whilst Russell's viper venom (RVV) can be used to assay clotting factors V, VII, X and lupus anticoagulants (LA). Activators from the taipan, Australian brown snake and saw scaled viper have also been used to assay LA. Protein C (PC) and activated PC (APC) resistance can be measured by means of RVV, Protac (from Southern copperhead snake venom) and STA-Staclot (from Crotalus viridis helleri) whilst von Willebrand factor can be studied with Botrocetin (Bothrops jararaca). Finally, snake venom C-type lectins and metalloproteinase disintegrins are being used to study platelet glycoprotein receptors and show great potential for use in the routine coagulation laboratory. PMID- 11910188 TI - Procoagulant proteins from snake venoms. AB - Several procoagulant proteins from snake venoms have been isolated and characterized. They are either serine proteinases or metalloproteinases, which activate specific zymogens of coagulation factors and initiate the coagulation cascade. These procoagulant proteins are useful in treating various thrombotic and hemostatic conditions and contribute to our understanding of molecular details in the activation of specific coagulation factors. Recent studies have shown that the prothrombin activators with serine proteinase activity are structurally and functionally similar to mammalian coagulation factors. Their structural studies should provide us insight into prothrombinase complex formation. Here, we present an overview of snake venom procoagulant factors. PMID- 11910189 TI - Snake venom activators of factor X: an overview. AB - Activators of blood coagulation factor X have been described in the venom of many snake species belonging to the genus Viperidae and Crotalidae as well as from a few Elapid species. Based on the structural and functional properties of purified activating principles, factor X activators are either metalloproteases or serine proteases. The best known activator is RVV-X from Russell's viper (Daboia russelli), a metalloprotease consisting of a heavy chain containing the catalytic domain and two light chains which share homology with C-type lectins and which are thought to exert a regulatory function in the Ca(2+)-dependent activation of factor X. This activator is also one of the best examples of the use of exogenous activators in coagulation research and in addition it is used in many diagnostic research kits. In this paper, an overview is given of the structural and functional properties of snake venom factor X activators thus far described in the literature. PMID- 11910190 TI - Snake venom prothrombin activators homologous to blood coagulation factor Xa. AB - We have recently determined the complete amino acid sequence of trocarin, a group D prothrombin activator from the venom of Tropidechis carinatus (Australian rough scaled snake). This proteinase is both functionally and structurally similar to mammalian blood coagulation factor Xa. It shows approximately 70% homology and possesses the characteristic Gla domain, two EGF domains and serine proteinase domain. To examine structure-function relationships, we generated a molecular model of trocarin based on a human factor Xa des-Gla crystal structure (1xka) as template. Based on known sites of interaction between mammalian factor Xa, factor Va and prothrombin, structure-function relationships of trocarin were explored. Unlike factor Xa, trocarin is glycosylated and has a large carbohydrate moiety at the entrance to its active site pocket. This might contribute to differences observed in the kinetics of hydrolysis of synthetic substrates by trocarin as compared to human factor Xa. A Ca(2+)-binding loop present in the heavy chain of factor Xa also seems to be lost in trocarin. In addition to its role in hemostasis, factor Xa shows other biological effects, including inflammation via its interaction with effector protease receptor-1 (EPR-1). Interestingly, the EPR 1 recognition site is distinctly different in trocarin, the functional consequences of which are being investigated. PMID- 11910191 TI - Factor V activation and inactivation by venom proteases. AB - Blood coagulation factor V is a single-chain glycoprotein with M(r) = 330,000 which plays an important role in the procoagulant and anticoagulant pathways. Thrombin activates factor V into factor Va, a two-chain molecule which is composed of a heavy (M(r) = 105,000) and a light chain (M(r) = 71,000/74,000). Factor Va accelerates factor Xa-catalysed prothrombin activation more than 1,000 fold and under physiological conditions the cofactor activity of factor Va in prothrombin activation is down-regulated by activated protein C. Factor V can also be activated by a wide variety of snake venoms (e.g. from Vipera species, Naja naja oxiana, Bothrops atrox) and by proteases present in the bristles of a South American caterpillar (Lonomia achelous). Some venoms, notably of Vipera lebetina turanica and Lonomia achelous, contain proteases that are able to inactivate factor V or factor Va. Venom factor V activators are excellent tools in studying the structure-function relationship of factor V(a) and they are also used in diagnostic tests for quantification of plasma factor V levels and for the screening of defects in the protein C pathway. In this review, the structural and functional properties of animal venom factor V activators and inactivators is described. PMID- 11910192 TI - Molecular basis for the partition of the essential functions of thrombin among snake venom serine proteinases: the case of thrombin-like enzymes. AB - Thrombin is a mammalian serine proteinase that plays a prominent role in the maintenance and regulation of hemostasis through its interaction with various substrates and/or ligands. The venoms of several snakes contain glycosylated serine proteinases that have been recognized to possess one or more of the essential activities of thrombin on fibrinogen (Fg) and/or platelets. These proteinases share about 60% sequence identity. One class of snake venom serine proteinases are those known as thrombin-like (TLE), named after their ability to directly clot Fg in order to preferentially produce fibrinopeptide A, fibrinopeptide B or both. To understand the molecular basis of this phenomenon, the corresponding amino acid sequences and molecular structures need to be analyzed. Given the absence of experimentally determined tertiary structures of snake venom, TLEs, three-dimensional molecular models should prove useful in this context. Towards this goal, we obtained models of snake venom TLEs that used TSV PA as template, TSV-PA being the only snake venom serine proteinase whose crystal structure is known to date. Along with a comparative sequence analysis the models contribute to the identification and description of thrombin-homologous or alternative binding sites, helping thus to understand differences in specificity. PMID- 11910193 TI - Effects of lopap on human endothelial cells and platelets. AB - Severe consumption coagulopathy has been detected in rats after Lopap (a prothrombin activator from Lonomia obliqua caterpillar bristles) infusion and in humans after accidental contact with L. obliqua bristles. However, platelet count and antithrombin (AT) levels were only modestly affected, suggesting that a different form of blood coagulation activation may be involved in this hemorrhagic syndrome. Here we describe that Lopap had no effect on aggregation of washed human platelets induced by several agonists, suggesting that it might not impair platelet function in vivo. AT was able to inhibit the amidolytic activity of thrombin generated by incubation of Lopap with prothrombin in a purified system, which may be different from that generated by the prothrombinase complex in vivo. The surface expression of both ICAM-1 and E-selectin but not of VCAM-1 was upregulated by Lopap in cultured HUVEC, suggesting that it may behave differently from other mediators, such as thrombin and tumor necrosis factor alpha. PMID- 11910194 TI - Protein C activators from snake venoms and their diagnostic use. AB - Proteinases converting the zymogen protein C (PC) of vertebrates into activated PC have been detected in several snake venoms. Most PC activators have been purified from venom of snake species belonging to the genera of the Agkistrodon complex. Unlike the physiological, thrombin-catalyzed PC activation reaction which requires thrombomodulin as a cofactor, most snake venom activators directly convert the zymogen PC into the catalytically active form which can easily be determined by means of coagulation or chromogenic substrate techniques. Due to this feature, the fast-acting PC activator Protac from Agkistrodon contortrix contortrix (southern copperhead snake) venom has found a broad application in diagnostic practice for the determination of disorders in the PC pathway. Recently, screening assays for the PC pathway have been introduced, based on the observation that the PC pathway is probably the most important physiological barrier against thrombosis. PMID- 11910195 TI - Interaction of bothrojaracin with prothrombin. AB - Bothrojaracin (BJC) is a 27-kD protein from Bothrops jararaca venom that interacts with alpha-thrombin (K(D) = 0.7 nM) through both anion-binding exosites I and II. Recently, it has been shown that BJC interacts with the exosite I precursor (proexosite I) on human prothrombin (K(D) = 75 nM), forming a 1:1 Ca(2+)-independent noncovalent complex with the zymogen. Complex formation was associated with inhibition of zymogen activation by Oxyuranus scutellatus venom. In addition, BJC strongly decreased the prothrombin activation by factor Xa only in the presence of factor Va. A similar effect was observed in the presence of phospholipids, suggesting that BJC specifically inhibits the interaction of prothrombin with factor Va. It is proposed that BJC has two independent mechanisms for anticoagulation: (1) inhibition of exosite-I-dependent activities on alpha-thrombin, and (2) inhibition of prothrombin activation through interaction with proexosite I. PMID- 11910196 TI - Anticoagulant venom and mammalian secreted phospholipases A(2): protein- versus phospholipid-dependent mechanism of action. AB - Some venom and mammalian-secreted phospholipases A(2) (sPLA(2)) have been described to exert an anticoagulant effect. This review will discuss and compare phospholipid-dependent versus protein-dependent mechanisms of action of these sPLA(2) on the coagulation cascade. The importance of venom proteins, and of the study of their pharmacological effects, to explore the physiological functions of homologous mammalian proteins is also pointed out. PMID- 11910197 TI - Lonomia genus caterpillar envenomation: clinical and biological aspects. AB - Persons who have been in contact with Lonomia achelous or Lonomia obliqua caterpillars present external and internal bleeding and opening of recently healed wounds. Hematological tests show normal platelet count, prolonged prothrombin time, activated partial thromboplastin time and thrombin time, totally corrected by normal plasma. Decreased fibrinogen (Fg), factor (F) V, FXIII, plasminogen and alpha(2)-antiplasmin with increased FVIII: C, von Willebrand factor, Fg degradation products and D dimers. Tissue plasminogen activator, plasminogen activator inhibitor and protein C varied. In L. achelous biological fluids, compounds with anticoagulant or procoagulant properties have been identified. In L. obliqua bristle extracts, mainly procoagulant activities have been identified. Subcutaneous injections of L. achelous crude extracts and a semipurified fraction reduce Fg, plasminogen and FXIII in rabbits. Intravenous injections of a very purified fraction of L. achelous in rabbits produce lysis of preformed thrombi, a decrease of Fg, plasminogen, alpha(2)-antiplasmin, FXIII and inhibition of postthrombolytic thrombus growth. Subcutaneous injections of L. obliqua bristle extracts prolong prothrombin time and activated partial thromboplastin time and reduce FXIII. Intravenous injections of crude bristle extract and a purified fraction of L. obliqua induce disseminated intravascular coagulation. PMID- 11910198 TI - Identification of anticoagulant activities in salivary gland extracts of four horsefly species (Diptera, tabanidae). AB - Anticoagulant activities against the extrinsic and intrinsic coagulation pathways were identified in salivary gland extracts (SGE) prepared from four tabanids (Hybomitra muehlfeldi, Tabanus autumnalis, Haematopota pluvialis, Heptatoma pellucens). All extracts prolonged human plasma clotting time in a dose-dependent manner and inhibited thrombin activity in the chromogenic substrate assay. Horsefly SGE did not inhibit factor Xa. Partial purification of SGE proteins using reversed-phase high-performance liquid chromatography revealed species specific differences in the elution profiles and range of fractions with anticoagulant activities. PMID- 11910199 TI - Hyperinsulinemia in pre- and post-pubertal children born small for gestational age. AB - BACKGROUND: Reduced fetal growth is a potential risk factor for development of metabolic abnormalities in later life. The relationship between low birthweight and impaired glucose tolerance, type 2 diabetes and insulin resistance in adulthood has been well documented. PURPOSE: Assuming that fetal undernutrition is associated with insulin resistance in middle age, we elected to study whether this process may already be present in young adults and adolescents born small for gestational age (SGA). SUBJECTS AND METHODS: Children born in Vall d'Hebron Hospital Infantil, Barcelona, between 1986 and 1989 and between 1978 and 1983 with birthweights below the third centile for the local standard values, were invited to participate in the present study. Of those, 51 (22 girls and 29 boys) were pre-pubertal with 9.4 +/- 0.2 years of age and 49 (29 girls and 20 boys ) were post-pubertal, with 17.3 +/- 0.3 years of age. All patients underwent a standard, 2-hour oral glucose tolerance test. Insulin and glucose responses were compared with our previously published data in control children with normal birthweight. RESULTS: The insulin response at 30 min after glucose load was significantly higher (p < 0.001) in pre- and post-pubertal girls and boys formerly SGA than in controls. In addition, the girls also had a higher insulin response at 60 and 120 min. Mean serum insulin (MSI), the area under the insulin curve during the glucose challenge, was statistically increased in pre- and post pubertal boys and girls born SGA when compared to controls. CONCLUSION: The presence of high insulin levels after an oral glucose challenge in children and adolescents born SGA might be considered as an early marker of subsequent insulin resistance in adulthood. Furthermore, our population offers the opportunity to study the natural course of hyperinsulinemia and its outcome. Follow-up of this cohort may be helpful in distinguishing a subset of young children and adolescents in whom therapeutic intervention could be done. PMID- 11910201 TI - Thyroid hormone levels during a 3-week professional road cycling competition. AB - AIMS: The aim of this investigation was to examine the thyroid hormone levels of professional cyclists during a 3-week stage competition (Vuelta a Espana 1998). METHODS: The study population was made up of 16 male cyclists from two world leading professional teams. Four blood samples were drawn (between 07:00 and 09:00 a.m.) from each participant before and at the end of the 1st, 2nd and 3rd weeks of competition. 3,5,3'(-Triiodothyronine (T(3)), free T(3) (FT(3)), thyroxine (T(4)), free T(4) (FT(4)) and thyroid-stimulating hormone (TSH) were determined in each blood sample by radioimmunoassay. RESULTS: Serum T(4), FT(4) and FT(3) levels showed a significant increase (p < 0.05) by the last week of competition while concentrations of TSH and T(3) remained unchanged. CONCLUSION: In conclusion, 3 weeks of competition provokes changes in basal thyroid hormone concentrations in professional road cyclists. PMID- 11910200 TI - Effect of near physiologic insulin therapy on hypoglycemia counterregulation in type-1 diabetes. AB - OBJECTIVES: The aim of this study was to examine hormonal counterregulation during insulin-induced hypoglycemia in type-1 diabetic patients during long-term near normoglycemic insulin therapy and intensive clinical care. METHODS: Type-1 diabetic patients (age 35.3 +/- 2 years, body mass index 22.8 +/- 1 kg x m(-2), mean diabetes duration 13.6 (11-17 years), mean HbA1c during the last year 6.6 +/ 0.1%) and nondiabetic subjects were studied during (0-120 min) and after (120 240 min) hypoglycemic (3.05 mmol/l) hyperinsulinemic (approximately 330 pmol/l) clamp tests. RESULTS: During hypoglycemia peak plasma concentrations of glucagon (199 +/- 16 vs. 155 +/- 11 ng/l, p < 0.05), epinephrine (4,514 +/- 644 vs. 1,676 +/- 513 pmol/l, p < 0.001), norepinephrine (2.21 +/- 0.14 vs. 1.35 +/- 0.19 nmol/l, p < 0.01) and cortisol (532 +/- 44 vs. 334 +/- 61 nmol/l) were reduced in the diabetic patients. Plasma lactate did not change from baseline values (0.51 +/- 0.06 mmol/l) in diabetic but doubled in healthy subjects (1.13 +/- 0.111 mmol/l, p < 0.001 vs. control). During the posthypoglycemic recovery period plasma concentrations of free fatty acids were higher in diabetic patients at 240 min (1.34 +/- 0.12 vs. 2.01 +/- 0.23 mmol/l, p < 0.05). CONCLUSION: Despite long term near physiologic insulin substitution and the low incidence of hypoglycemia, hormonal hypoglycemia counterregulation was impaired in type-1 diabetic patients after a diabetes duration of more than 10 years. PMID- 11910202 TI - Alteration of endothelin-1 concentration in STZ-induced diabetic rat nephropathy. Effects of a PGI(2) derivative. AB - BACKGROUND: Recently, an endothelin (ET-1) with a potent vasoconstrictive activity and stimulative activity of vascular muscular cell growth was discovered and blood ET-1 levels were higher in diabetic patients than in healthy subjects, suggesting that high ET-1 levels assist development and progression of diabetic microangiography. METHODS: We examined renal function, and serum and tissue ET-1 levels in streptozotocin (STZ)-induced diabetic rats treated with a prostaglandin (PG) I(2) derivative to investigate the effect of PGI(2) in diabetic vascular disturbance. RESULTS: Renal weight, urinary albumin, urinary N-acetyl-beta,D glucosaminidase (NAG) and serum ET-1 levels increased in STZ-induced diabetic rats, and a tendency to increase in renal tissue ET-1 levels was observed. Furthermore, electron-microscopic findings in the kidneys showed mesangial cell proliferation and mesangial matrix expansion which might be caused by diabetic nephropathy. The PGI(2) derivative reduced urinary albumin and NAG levels in STZ induced rats. It was considered, therefore, that the PGI(2) derivative is effective in diabetic nephropathy. As the PGI(2) derivative also reduced renal tissue ET-1 levels, improvement of diabetic nephropathy partially was considered to result from the reduction of renal tissue ET-1 levels. CONCLUSION: In STZ induced rats, increased serum ET-1 levels and a tendency to increase in renal tissue ET-1 levels were associated with increases in urinary albumin and NAG levels, and these levels were decreased by a PGI(2) derivative. These findings suggested that increased ET-1 concentrations assist development and progression of diabetic nephropathy, especially diabetic microangiopathy, and the PGI(2) derivative may be effective for inhibition of diabetic microangiopathy mediated by reduction of ET-1 concentrations. PMID- 11910203 TI - Age-related prevalence of platelet-associated immunoglobulins G in nonthrombocytopenic patients with autoimmune thyroid disease and autoimmune polyglandular syndrome. AB - OBJECTIVE: The prevalence of platelet-associated IgG (paIgG) in nonthrombocytopenic patients with autoimmune thyroid disease (AITD) alone or associated with autoimmune polyglandular syndrome (APS) has been studied. SUBJECTS: A total of 164 individuals were enrolled in this study: 81 patients with AITD alone, 33 patients with APS, and 50 healthy controls. RESULTS: The presence of paIgG was recorded in 41 of 81 patients with AITD (51%) as compared with 2 of 50 control subjects (4%, p < 0.0001). The prevalence of paIgG in patients with APS was higher even when compared with patients with AITD alone (25/33, 76%; p = 0.02). The presence of paIgG was not related to the functional thyroid parameters. The prevalence of paIgG was higher in the older than in the younger patients (75 vs. 47%, p = 0.0037). CONCLUSIONS: The results indicate that the prevalence of paIgG in patients with AITD is higher than previously thought, namely in elderly patients and in patients with APS, and not related to the thyroid function. PMID- 11910204 TI - Kinetics and effect of percutaneous administration of dihydrotestosterone in children. AB - BACKGROUND: Percutaneous administration of dihydrotestosterone (DHT) has been successful in promoting phallic growth in infants and children with 5 alpha reductase deficiency raised as males. We investigated whether percutaneous administration of DHT is similarly effective in patients with micropenis due to alternative diagnoses. METHODS: Six patients (age range 1.9-8.3 years) with micropenis of variable etiology were studied prospectively. 2.5% DHT gel was applied to the phallus once daily at a dose of 0.15-0.33 mg/kg body weight. Serum DHT concentrations were measured at 0, 2, 4, 8, 12 and 24 h following application of DHT gel. RESULTS: Peak DHT concentrations were attained within 2-8 h after application of the gel and subsequently remained within the normal adult range in all but 1 patient, who had received the lowest dose of 0.15 mg/kg. An increase in phallic growth, ranging from 0.5-2.0 cm, was achieved after 3-4 months of treatment in all patients whose DHT concentrations were maintained within adult range. CONCLUSION: Percutaneous administration of DHT in a dose of 0.2-0.3 mg/kg once daily for a period of 3-4 months may be useful in the management of patients with testosterone biosynthetic defects, who have sufficient masculinization to warrant male sex assignment, or in patients with micropenis prior to reconstructive surgery. PMID- 11910205 TI - Molecular analysis of X-linked ichthyosis in Japan. AB - BACKGROUND: X-linked ichthyosis (XLI) is an inherited skin disorder caused by a deficiency of steroid sulfatase (STS). The gene and protein of STS were examined in 19 Japanese patients with XLI. RESULTS: In Western blotting analysis, no cross reacting peptide was detected in the patients' placenta, although a single band (63 kD) corresponding to STS in a normal subject was observed. Southern blotting was performed using EcoRI digests of cellular DNA from 13 XLI patients and full length human STS cDNA as a probe. Normal males had bands of 20, 15, 10, 9.0, 6.1, 4.2, 2.6, and 1.5 kb. Twelve of the 19 patients had only 20- and 1.5-kb bands. Only one patient had the same band pattern as that of normal males. The STS gene was analyzed by PCR in 6 of the 19 patients. PCR amplification products were sequenced to analyze the STS gene. Two cases with one-base change in the STS gene and variation in amino acids H444R and E560P were found. Mutant STS cDNA was transfected into COS-1 cells and the STS enzyme activity was assayed. The enzyme activities were less than the minimum detection value of the detection system. CONCLUSIONS: These results suggest that XLI is mainly caused by an extensive deletion of the STS gene and that the PCR method is useful for detection of STS point mutations. PMID- 11910206 TI - The European Training Syllabus in Paediatric Endocrinology and Diabetes. PMID- 11910208 TI - Ipsilateral thalamic stimulation after thalamotomy for essential tremor. A case report. AB - We report a patient with severe essential tremor who was treated with thalamic stimulation ipsilateral to a prior thalamotomy. Thalamotomy performed 30 years prior to stimulator implantation provided tremor reduction for one year before the tremor recurred. An electrode lead was implanted in the thalmaic nucleus ventralis intermedius (Vim) with nearly complete control of his tremor with sustained benefit over an 18-month follow-up period. Vim thalamic stimulation is an effective treatment option for recurrent tremor in patients who have undergone ipsilateral thalamotomy. PMID- 11910209 TI - Temporal sequence of response to unilateral GPi pallidotomy of motor symptoms in Parkinson's disease. AB - The present study was performed to determine the temporal sequence of the response to unilateral MRI/microelectrode-guided pallidotomy of each cardinal symptom in Parkinson's disease (PD). For this purpose, we performed a quantitative assessment of motor functions in 19 patients with PD at several time points up to 6 months following surgery. We here report that although all the motor signs were significantly improved 6 months after pallidotomy, the temporal sequence of tremor response was different from those of other symptoms. PMID- 11910210 TI - Continuous intrathecal clonidine administration for the treatment of neuropathic pain. AB - In many cases, the treatment of neuropathic pain by intrathecal opioids fails to meet expectations. In a trial involving 10 patients, the intrathecal administration of clonidine combined with opioids in the treatment of chronic pain was introduced in our department for the first time. Eight patients with neuropathic pain syndromes were subjected to a continuous intrathecal clonidine application in addition to intrathecal morphine. At an average dose of 44 microg clonidine/day, a 70-100% reduction in pain was achieved. Residual non-neuropathic pain in 4 of 8 patients was successfully treated with clonidine and low doses of opioids. On the basis of the results achieved so far, we recommend that clonidine should be routinely tested for intrathecal drug administration, especially in patients with a prominent neuropathic pain component. PMID- 11910211 TI - Personal-computer-based system for three-dimensional anatomic-physiological correlations during stereotactic and functional neurosurgery. AB - This paper describes the automatic three-dimensional (3D) graphic possibilities that are supplied by the Neurosurgical Deep Recording System (NDRS) to facilitate anatomic-physiological targeting during stereotactic and functional neurosurgery using depth recording. This software has been developed to substitute the complex electronic equipment ordinarily used for deep brain electrical recording, display and processing by a personal computer. It may also help to improve on-line graphic analysis, automatic management of the recorded information and flexibility to implement different forms of signal analysis. It can automatically show a 2D or 3D representation of the electrode track, with the electrophysiological findings superimposed as well as the corresponding sagittal, coronal and axial views of a brain atlas using automatic scaling. The NDRS has already successfully been applied during more than 300 neurosurgeries in Spain and Cuba, enabling improved targeting accuracy and safety. PMID- 11910214 TI - American Journal of Therapeutics. PMID- 11910212 TI - Factors influencing the application accuracy of neuronavigation systems. AB - OBJECTIVE: The overall accuracy of neuronavigation systems may be influenced by (1) the technical accuracy, (2) the registration process, (3) voxel size and/or distortion of image data and (4) intraoperative events. The aim of this study was to test the influence of the registration and imaging modality on the accuracy. METHODS: A plexiglas phantom with 32 rods was taken for navigation targeting. Sixteen fiducials were attached to the surface of the phantom forming two different attachment patterns (clustered vs. diffusely scattered). This model was scanned by MRI and CT (1-mm slices). Registration was performed using different numbers and attachment patterns of the fiducials. Using CT or MRI, the localization error was measured in image space as the Euclidean distance between targets defined in image space and those detected in the physical space. Accuracy was measured with two commercial systems, the Zeiss MKM and the StealthStation. RESULTS: The mean localization error varied between 1.59 +/- 0.29 mm (MKM, 8 scattered fiducials, CT scanning) and 3.86 +/- 2.19 mm (MKM, 4 clustered fiducials, MRI). The worst localization error was 9.5 mm (MKM). In case of an optimal registration, the 95th percentile for the localization error was 2.2 (MKM) and 2.75 mm (StealthStation). The imaging modality has only minor influence on the localization error, with CT increasing accuracy minimally. Both the fiducial number and the attachment pattern critically influence the localization error: 8 fiducials and a generalized attachment pattern increase the accuracy significantly. No correlation between the calculated registration accuracy and the measured localization accuracy was found. CONCLUSION: The application accuracy of different neuronavigation systems critically depends on the registration. The calculated registration accuracy provided by the system does not correspond to the localization error found in reality. The accuracy of frameless neuronavigation systems is comparable to that of classical frame-based stereotactic devices. PMID- 11910215 TI - Therapeutic effect of using a long-pulsed alexandrite laser system with a cooling device for epilation in reconstructive surgery of auricular malformations. AB - Hair growth after reconstructive surgery for auricular malformations including microtia and full-thickness skin grafts poses serious problems for patients. Currently, the author and his colleagues use a long-pulsed alexandrite laser system with a cooling device to remove hair from hairy regions before and after reconstructive surgery of auricular malformations. They achieve good results with approximately two to five treatments, and report the details here. Using their protocols there were no adverse reactions such as ulceration, scarring, pigmentation, or depigmentation. The use of this system may change the technique of auricle treatment because of the advantages it offers, such as low invasiveness, high effectiveness, and safety. PMID- 11910216 TI - A personal technique: mammaplasty with J scar. AB - Mastopexy and reduction mammaplasty techniques have evolved with time, pursuing the aim of an effective and reliable technique that produces a well-shaped breast and reduces the amount of scarring. The authors believe that the L mammaplasty achieves the best results in terms of a short scar and a good, stable shape. They present their technique of a modified L mammaplasty with a resulting scar in the shape of a J, which implies a central breast resection with the nipple-areola complex transposed on a superior pedicle. The correct execution of the preoperative markings and the shaping of the gland tissue are mandatory to obtaining the desired result. The technique has been used during the past 7 years on 326 patients, providing satisfactory results with short scars and virtually no complications. PMID- 11910217 TI - Correction of severe secondary cleft lip nasal deformity using a composite graft: current approach and review. AB - In the case of a severe tissue deficiency with a secondary cleft lip nasal deformity, a composite graft can be useful for columellar lengthening or to create symmetrical nostrils. The current study used composite grafts to correct secondary cleft lip nasal deformities with a severe tissue deficiency or severe nostril asymmetry. A total of 19 patients who were born with complete cleft type were operated between 1995 and 1999. Of these patients, 10 were men and 9 were women, and the age distribution was 7 to 35 years old. In 9 patients with unilateral cleft lip nasal deformities and in 6 patients with bilateral cleft lip nasal deformities, columellar lengthening was performed using a composite graft taken from the helix in 14 patients and the contralateral alar rim in 1 patient. In 4 patients with severely asymmetrical nostrils resulting from a short alar rim in unilateral cleft lip nasal deformities, the ear helix was used in 2 patients, whereas in the other two patients, the alar rim of the unaffected side was transferred to the affected side to create a symmetrical nostril by reducing the length of the ala on the unaffected side. The follow-up period ranged from 1 to 3 years, and results were as follows: Four days after the graft, the composite tissue exhibited a pinkish color, and complete survival was confirmed after 7 days. The absorption rate was approximately 10% and the color mismatch became minimal with time. Composite tissue from the ear was found to be useful for full layer reconstruction of the ala and columella because of its stiffness, thin nature, and similarity. Composite tissue from the alar rim on the contralateral side was also determined to be a good material for full-layer reconstruction of the deficient ala. PMID- 11910218 TI - Articulation disorders associated with maxillary growth after attainment of normal articulation after primary palatoplasty for cleft palate. AB - Little attention has been given to the connection between maxillary growth and the emergence of articulation disorders among cleft palate patients who previously successfully attained normal articulation after undergoing primary palatoplasty. Follow-up assessment of 22 surgically treated unilateral cleft lip and palate patients who attained normal articulation before school entry revealed that, during mixed dentition, half the patients exhibited palatalized articulation and half retained normal articulation. To determine the possible effect on articulation of growth changes in dental arch dimensions, a comparison was made of deciduous dentition and mixed dentition oral cavity measurements. Indications are that palatalized articulation occurs in patients with a smaller anterior palatal volume, the presence of linguoversion teeth, and poorer growth ability. Results reveal the importance of early management of dental arch dimensions, and the need for periodic assessment and management of articulation even for patients who attain normal articulation subsequent to primary palatoplasty. PMID- 11910219 TI - Shur-clens: an agent to remove silicone gel after breast implant rupture. AB - Removal of silicone gel from surrounding tissues after implant rupture is difficult. Local inflammation, infection, and silicone granulomas warrant thorough removal of the silicone gel. Shur-Clens (20% solution of the surfactant poloxamer 188), povidone-iodine, and saline are agents that are used to aid in the removal of silicone gel from tissue. The purpose of this study was to compare the efficacy of silicone gel removal by these three agents in vitro. Shur-Clens, povidone-iodine, and saline were compared as solvents for silicone gel. Four weight increments of silicone gel (0.02 g, 0.04 g, 0.06 g, and 0.08 g) were placed on glass slides. These slides were placed in separate beakers containing 40 ml test solution. The slides were soaked for 1 minute with gentle agitation. The slides were removed, rinsed gently with de-ionized water, and placed in a vacuum desiccator to dry. The slides were weighed to determine the amount of silicone removed after soaking in the solution. Analysis of variance was used to determine the significance between the three solvents. The percentages of silicone gel removed for the four weight increments (0.02 g, 0.04 g, 0.06 g, and 0.08 g) in saline were 5.6%, 2.9%, 2.1%, and 5.8%, respectively. In povidone iodine solution, the percentages were 18.9%, 25.4%, 28.8%, and 51.9%. In Shur Clens, the percentages were 31.3%, 43.0%, 63.5%, and 79.9%. The greater percentage of silicone gel removed by Shur-Clens was significant compared with the other solutions (p < or = 0.05). Shur-Clens was shown to be a more effective solvent for removal of silicone gel in vitro. This enhanced efficacy is a result of the fact that Shur-Clens contains 20% of the surfactant poloxamer 188. The authors' clinical experience with 7 patients who underwent ruptured silicone breast implant removal demonstrated the superiority of Shur-Clens. Shur-Clens is a surfactant cleanser that is widely available, is inexpensive, and has a good safety profile. They propose the use of Shur-Clens to clean silicone gel spillage to decrease local complications resulting from residual silicone gel. PMID- 11910220 TI - Surgical treatment of painful neuroma in medial antebrachial cutaneous nerve. AB - The branches of the medial antebrachial cutaneous nerve (MACN) are located at the medial site of the elbow. The MACN, especially the posterior branches, may be injured or transected during cubital tunnel surgery or other medial approaches to the elbow. Damage to the nerve can cause a neuroma, which leads to disabling pain and restriction of elbow movement. The initial treatment of the neuroma is nonsurgical, and includes local massage, desensitization, physiotherapy, and systemic medication. If after 6 months of these nonsurgical treatments there is no improvement, surgery is indicated. The authors report their experience with 12 patients treated surgically for painful neuroma by high resection of the proximal end or its implantation into the triceps muscle. After surgery there was a high success rate of pain relief and functional improvement in both elbow movement and handgrip strength. PMID- 11910221 TI - Versatility of the free anterolateral thigh flap for reconstruction of soft tissue defects: review of 140 cases. AB - From August 1995 to June 1999, 140 free anterolateral thigh (ALT) flaps were transferred to reconstruct a variety of soft-tissue defects. The size of ALT flap ranged from 10 to 33 cm in length and 4 to 14 cm in width. Based on the anatomic variations of the perforators, the blood supply to the skin island came from the septocutaneous perforators only in 19 patients (13.6%), arising from the descending or transverse branch of the lateral circumflex femoral artery (LCFA), or originating directly from LCFA. The other flaps were supplied by musculocutaneous perforators that were elevated as a true perforator flap via intramuscular dissection (N = 34, 24.3%), or used a cuff of vastus lateralis muscle for added bulk (N = 87, 62.1%). The overall success rate was 92% (129 of 140). After a 2-year follow-up, all flaps have healed unevenffully and donor thigh morbidity is minimal. Anatomic variations must be considered if the ALT flap is to be used safely and reliably. PMID- 11910222 TI - Microsurgical free flap transfer to amputation sites: indications and results. AB - A series of microsurgical free flap reconstructions to amputation stumps of the upper as well as the lower extremities was reviewed in 7 male and 2 female patients. Indications included preservation of length after trauma in 6 patients and cure of local infection in 2 patients. In 1 patient an extensive defect after resection of a recurrent shoulder sarcoma required use of a complete arm fillet free flap for tumor reconstruction. Microvascular free flaps used included four scapular flaps, two fillet flaps from the amputated extremity, one anterolateral thigh flap, and one lateral arm flap. Seven of 9 patients were fitted with a prosthesis and underwent occupational therapy resulting in ambulatory and improved functional status. Microvascular reconstruction is indicated in emergency settings as well as for elective reconstruction of amputation sites. Using uninjured "spare parts" of the amputated extremity should be considered. Elective reconstruction is performed preferably with free flaps based on the subscapular vascular system. PMID- 11910223 TI - Endoscopic harvest of four muscle flaps: safe and effective techniques. AB - The recent explosion of endoscopic techniques in plastic surgery has led to the successful harvest of a number of useful muscle flaps. The gracilis, rectus femoris, external oblique, and gastrocnemius muscles can all be harvested safely and reproducibly using endoscopic techniques. The aim of this study was to identify a safe and effective technique for endoscopic muscle flap harvest. Harvesting the gracilis muscle as a free flap and the gastrocnemius as a pedicle flap lends themselves best to the use of endoscopic techniques. PMID- 11910224 TI - Digital photography as a means of enhancing interconsultant communication in oncological cutaneous surgery. AB - Plastic surgeons have associated themselves closely with photography almost since the inception of medical photography. Photographs are used for documentation of medical records and comparison of pre- and postoperative status. Digital photography allows instant picture viewing and printing as well as Web-based mailing and, as a result, it facilitates interconsultant communication between the dermatologist, surgeon, and pathologist during the surgical treatment of cutaneous neoplasms. Moreover, it enhances patient monitoring during long-term follow-up. PMID- 11910225 TI - Tragic case of a dog bite in a young child: the dog stands trial. AB - The authors present the tragic case of an 18-month-old child who was bitten by a dog, causing amputation of the forearm and substantial damage to the cutaneous muscle on his back, shoulder, thorax, and neck. A free latissimus dorsi flap was performed to preserve the humerus from which the periosteum had been torn away. A series of cutaneous expansions were then undertaken to graft skin back onto the back, the armpit, and the shoulder stump, to allow for a mechanical prosthesis. A study of the literature on this subject proves that dog bites are more frequent and serious (sometimes even fatal) in young children than in adults. In view of the current legislation, it would seem that the public health authorities are doing little to resolve this distressing problem. PMID- 11910226 TI - Use of a venous flap from an amputated part for salvage of an upper extremity injury. AB - The authors describe a patient in whom a large arterialized venous flap was harvested from a nonreplantable part after partial hand amputation. A 9 x 6-cm segment of dorsal hand skin was transplanted acutely in an artery-vein-vein fashion to cover exposed bone, joints, and reconstructed tendons. The flap provided durable coverage, and at 1 year the patient regained 94% total active motion at the index finger and 99% total active motion at the long finger. Salvage of component parts such as a venous flap and extensor tendons avoided additional procedures for coverage and staged tendon reconstructions. PMID- 11910227 TI - Acute renal failure during dextran-40 antithrombotic prophylaxis: report of two microsurgical cases. AB - Dextran is often used to enhance the successful outcome of microvascular transplantations. So far, some 60 cases of dextran-induced acute renal failure have been reported. Among them, only one has been reported as an adverse reaction to antithrombotic prophylaxis during microvascular surgery. To stress that dextran prophylaxis can be a serious threat to microsurgical patients, the authors report on two more such patients and discuss how to prevent and treat this condition. Hypovolemic patients of older age are at risk, as are patients with cardiovascular diseases, renal artery stenosis, and preexisting renal insufficiency. Dextran should be discontinued promptly in patients with decreased urinary output or with high urine specific gravity. Dextran-induced acute renal failure may be severe and it may last several days. Plasmapheresis is the therapy of choice and, usually, diuresis and renal function resume shortly after such treatment. PMID- 11910228 TI - Proteus syndrome in adulthood. AB - Proteus syndrome is a very rare congenital condition comprising malformations and overgrowth of multiple sorts of tissue. It was described for the first time in 1979 and was termed Proteus syndrome in 1983. The authors describe a 37-year-old patient who was diagnosed initially as having Klippel-Trenaunay-Weber syndrome at the age of 10 years. The patient was operated for a major thoracic lymphatic malformation, which caused functional problems. The operation at older age for this large lymphatic malformation proved to be complex. In addition, the authors address the difficulties in diagnosing Proteus syndrome. Vascular malformations causing functional problems in adulthood require major surgical procedures with a high risk of postoperative complications. PMID- 11910229 TI - Mafenide acetate allergy presenting as recurrent chondritis. AB - Acute chondritis has a strong predilection for recurrence. Mafenide acetate has been implicated in causing reactions that mimic this condition; however, these hypersensitivity reactions lack fever, fluctuance, and pain. The authors report a case of mafenide acetate allergy presenting as recurrent chondritis in a patient who had previously been treated successfully for this condition. In this patient, the allergic response resolved within 3 days after cessation of mafenide acetate. If unappreciated, it may have led to unnecessary operative intervention. Therefore, auricular edema and erythema, without fever, fluctuance, and pain, must be recognized by surgeons as a possible mafenide acetate allergy and must be considered in the differential diagnosis for patients who present with recurrent acute suppurative chondritis. PMID- 11910230 TI - Intraoperative lymphatic mapping to treat groin lymphorrhea complicating an elective medial thigh lift. AB - Groin lymphoceles and lymphorrhea are a rare complication of medial thigh lift procedures. The author describes a case in which a very thin patient developed groin lymphorrhea after an uncomplicated medial thigh lift procedure. Initial treatment interventions, including edema control and the placement of a drain with surgical exploration, failed to control the lymphatic leak. Additionally, the onset of an infection and abscess formation complicated the treatment efforts. Using techniques well established in treating cutaneous malignancies, the lymphocele was treated successfully by identifying three separately damaged lymphatic channels with the use of intraoperative lymphatic mapping with blue dye. No drains were needed and the immediate cessation of lymph flow was noted. Using this novel adaptation of a well-known technique, the groin lymphocele was able to be repaired quickly and effectively with minimal morbidity and no evidence of recurrence to date. PMID- 11910231 TI - Two-stitch technique for distal shaft hypospadias repair. AB - The authors present a new technique for distal shaft hypospadias repair using a flip flap fashioned with only two stitches. In raising the flap, the tissues lateral and proximal to the flap are undermined but there is no undermining beneath the flap. The flap is sutured up to the glans with two 4-0 Vicryl sutures. No sutures are placed at the lateral edges of the flap to create the "tube" of the neo-urethra, and no postoperative urinary drainage is used. Thirty three children with distal shaft hypospadias underwent the "two-stitch" flip flap operation. The complications were one fistula and one case of urinary retention. The technique is an easy method for reconstructing distal penile hypospadias with a very low rate of complications and is suitable for an outpatient surgical setting. PMID- 11910233 TI - Re: Successful breast reconstruction with a perforator-to-deep inferior epigastric perforator flap. PMID- 11910234 TI - Adequate depth of excision for basal cell carcinoma of the nose. PMID- 11910235 TI - Basal cell carcinoma of the scalp 70 years after irradiation. PMID- 11910236 TI - Basal cell carcinoma metastatic to the parotid stroma. PMID- 11910237 TI - Unusual schwannoma of the buccal and temporal regions. PMID- 11910238 TI - Hemangioma in masseteric muscle. PMID- 11910240 TI - Cylindroma of the leg. PMID- 11910239 TI - Cylindroma of the foot. PMID- 11910241 TI - Posttraumatic lipogranuloma of the hand. PMID- 11910242 TI - Is rational flap selection and good surgical technique sufficient for treating pressure ulcers? The importance of psychology: a prospective clinical study. PMID- 11910243 TI - Bridging cleft palate. PMID- 11910244 TI - Efficacy of moist exposed burn ointment in the management of cutaneous wounds and ulcers: a multicenter pilot study. PMID- 11910245 TI - Cardiovascular effects of inhalational anesthetics. PMID- 11910246 TI - Contributing Authors. PMID- 11910248 TI - Preface. PMID- 11910247 TI - Cardiovascular effects of intravenous anesthetics. PMID- 11910249 TI - New drugs: fenoldopam mesylate, glycoprotein IIb/IIIa antagonists, and K+ATP channel agonists. PMID- 11910250 TI - General anesthesia versus regional anesthesia. AB - No distinct advantage is apparent between regional and general anesthesia when considering perioperative cardiac morbidity and mortality in peripheral vascular surgery. However, there is some evidence to support regional anesthesia over general anesthesia in an effort to optimize graft patency if the regional technique is extended into the postoperative period to provide neuraxial analgesia. An inadequate number of randomized, controlled trials have been conducted to determine whether regional or general anesthesia should be performed for carotid endarterectomy. The nonrandomized trials do support regional anesthesia by virtue of reductions in stroke, myocardial infarction, and death. A randomized, prospective trial is needed to verify these outcomes. The choice of technique does not appear to affect mortality in patients requiring hip fracture surgery, although Urwin et al. (29) reported less 1-month mortality in patients receiving regional anesthesia. General anesthesia has been associated with increased blood loss and thromboembolic complications in patients undergoing hip fracture repair. Epidural anesthesia has been shown to promote quicker return of bowel function postoperatively when the catheter has been sited at T12 or higher. Anastomotic breakdown in patients with epidural anesthesia/analgesia has rarely been reported. Most studies tend to show quicker return of bowel function when local anesthetics alone are administered epidurally. PMID- 11910251 TI - Transesophageal Echocardiography in the noncardiac surgical patient. PMID- 11910252 TI - Myocardial ischemia monitoring. PMID- 11910253 TI - Mast cells and stress--a psychoneuroimmunological perspective. PMID- 11910254 TI - The effects of antipsychotic-induced hyperprolactinaemia on the hypothalamic pituitary-gonadal axis. AB - Hyperprolactinaemia is commonly induced by antipsychotic medications that have dopamine-blockade as their main mechanism of action. The purpose of this study was to assess the effect of antipsychotic-induced hyperprolactinaemia on hypothalamic-pituitary-gonadal axis (HPG) function.HPG axis function was assessed in 67 consecutive outpatients who were diagnosed with schizophrenia and stabilized for a period of not less than 2 years on typical antipsychotic medication, by means of clinical history, relevant questionnaires and measurement of plasma prolactin, estradiol, progesterone, testosterone, LH, FSH, sex hormone binding globulin, and TSH levels. Normative laboratory data were used to assess whether hormone levels fell within the reference range for a normal population. There was a significant correlation between dose of medication and plasma prolactin levels for the total group (P<0.001). Prolactin levels were significantly negatively associated with sex hormone levels in females (P<0.05). Males taking antipsychotic medication had a mean prolactin level of 404.1m/IU and mean gonadotrophin and sex hormone levels that fell within normal limits. The results of this study indicate that neuroleptic-induced prolactin secretion is a dose-related side effect and, in females, the level of hyperprolactinaemia is correlated with the degree of suppression of the HPG axis. Women taking long-term prolactin-raising antipsychotic medications are likely to be hyperprolactinaemic and have an associated hypogonadal state. In males, prolactin levels remain within normal limits, but at the upper end, with no apparent disturbance of reproductive hormones. PMID- 11910255 TI - Would a switch from typical antipsychotics to risperidone be beneficial for elderly schizophrenic patients? A naturalistic, long-term, retrospective, comparative study. AB - Elderly chronic schizophrenia patients are particularly difficult to treat because of aging-related changes, cognitive impairment, and comorbid physical illness. This article describes a naturalistic, retrospective study of typical antipsychotic treatment versus risperidone for elderly psychotic inpatients. Fifty-one patients, mean age 72.7 + 5.9 years, mean disease duration 33.1 + 12.0 years, who met the DSM-IV criteria for schizophrenia or schizoaffective disorder were treated by risperidone (n = 26) or typical antipsychotic treatment (n = 25) during acute exacerbation and followed up for 18 months. Patients were rated using the clinical global impression (CGI) scale and positive and negative symptom scale (PANSS), and their body weight was recorded at baseline, 6 months, and 18 months. Both treatment groups improved on all rating scales at 18 months. Levels of decrease in PANSS positive and total scores were more prominent in patients treated with risperidone (p < 0.01 and p < 0.05, respectively). The change in CGI scores reached significance only after 18 months and was more pronounced in the risperidone group (p < 0.01). Anti-Parkinsonian medications were used more frequently in the typical antipsychotic group, whereas benzodiazepines were used more frequently in the risperidone group. Body mass index increased minimally after 18 months in the risperidone group (+ 0.3 kg/m2), whereas a larger (+ 1.1 kg/m2), albeit not statistically significant, increase was recorded in the typical antipsychotic group. Emergence of side effects was less frequent in patients treated with risperidone (4/26 vs. 16/25 patients, p < 0.01). The results of this study demonstrate that in elderly chronic schizophrenic patients, switching from typical antipsychotics to risperidone is effective and well tolerated. PMID- 11910256 TI - The safety and pharmacokinetics of quetiapine when coadministered with haloperidol, risperidone, or thioridazine. AB - The effects of haloperidol, risperidone, and thioridazine on the pharmacokinetics and side-effect profile of quetiapine were investigated in 36 patients with schizophrenia, schizoaffective disorder, or bipolar disorder in a single-center, two-period, multiple-dose, open-label, randomized trial. Over a one-to two-week period, quetiapine doses were escalated to 300 mg twice daily (bid). Patients were then treated for at least 7 days at the target quetiapine dose and subsequently entered into the combination therapy period, receiving haloperidol (7.5 mg, bid), risperidone (3 mg, bid), or thioridazine (200 mg, bid) for 8.5 days (after 3 days of dose escalation). Key assessments included the pharmacokinetics of quetiapine at steady state (area under the curve within a dosing interval [AUCtSS], maximum [CmaxSS], and minimum [CminSS] observed plasma concentrations, and oral clearance [Cl/f]), as well as the UKU Side Effect Rating Scale scores and safety evaluations. Neither risperidone nor haloperidol had significant effects on quetiapine pharmacokinetics. However, thioridazine produced statistically significant changes, decreasing the least squares means values of the AUCtSS, CmaxSS, and CminSS by 40%, 47%, and 31%, respectively, and increasing Cl/f by 68%. Increases in the following adverse events were noted during coadministration: somnolence (risperidone), insomnia and dry mouth (all three coadministered therapies), and dizziness (thioridazine). UKU side effect items that became worse in >or= 25% of patients during each coadministration period included sedation and increased sleep duration. Results of laboratory tests, electrocardiograms, and vital sign measurements revealed few clinically important changes. Clinical stability can be maintained with good tolerability during the transition from quetiapine monotherapy to periods of coadministration with haloperidol, risperidone, or thioridazine. Coadministration of either haloperidol or risperidone did not have any important effects on the steady-state pharmacokinetics of quetiapine. Thioridazine significantly increased the oral clearance of quetiapine. Increased doses of quetiapine may be necessary to control psychotic symptoms when thioridazine is coadministered with quetiapine. PMID- 11910257 TI - Serotonin and fluoxetine levels in plasma and platelets after fluoxetine treatment in depressive patients. AB - Depression is a mood disorder characterized by complex alterations of neurotransmitters such as serotonin, norepinephrine, and dopamine. In particular, there is substantial evidence of abnormalities in serotonin neurotransmission. Peripheral parameters of serotoninergic transmission, such as the 5 hydroxytryptamine content of plasma and platelets, have been used to identify biochemical alterations related to depression. In recent years, these parameters have also been used to examine the mechanism of action of antidepressive drugs such as the selective serotonin reuptake inhibitors. This study investigated the interaction between the plasma and platelet levels of fluoxetine and serotonin after fluoxetine administration to depressed patients. Twelve patients affected by major depression (according to the DSM-IV criteria) received a single oral dose of fluoxetine in the morning: 5 mg in the first 5 days, 10 mg from day 6 to day 10, and 20 mg from day 11 to day 40. Blood samples were collected at 0, 7, 10, and 24 hours after drug administration on the day 1 of fluoxetine 5 mg and on the 1st and the 30th day of fluoxetine 20 mg (days 11 and 40 of treatment, respectively). Plasma fluoxetine and serotonin levels increased after drug administration, reaching the highest levels on the 30th day of fluoxetine 20 mg. Fluoxetine levels were also detectable in platelets, with a time variation similar to plasma values. Platelet serotonin levels decreased after drug administration, and the lowest values were observed on the 30th day of fluoxetine 20 mg. PMID- 11910258 TI - Acute efficacy of fluoxetine versus sertraline and paroxetine in major depressive disorder including effects of baseline insomnia. AB - This study assessed whether fluoxetine, sertraline, and paroxetine differ in efficacy and tolerability in depressed patients and the impact of baseline insomnia on outcomes. Patients (N = 284) with DSM-IV major depressive disorder were randomly assigned in a double-blind fashion to fluoxetine, paroxetine, or sertraline for 10 to 16 weeks of treatment. Using the Hamilton Rating Scale for Depression (HAM-D) sleep disturbance factor score, patients were categorized into low (<4) or high (>or=4) baseline insomnia subgroups. Changes in depression and insomnia were assessed. Safety assessments included treatment-emergent adverse events (AEs), reasons for discontinuation, and AEs leading to discontinuation. In addition, AEs were evaluated within insomnia subgroups to determine emergence of activation or sedation. Depression improvement, assessed with the HAM-D-17 total score, was similar among treatments in all patients (p = 0.365) and the high (p = 0.853) and low insomnia (p = 0.415) subgroups. Insomnia improvement, assessed with the HAM-D sleep disturbance factor score, was similar among treatments in all patients (p = 0.868) and in the high (p = 0.852) and low insomnia (p = 0.982) subgroups. Analyses revealed no significant differences between treatments in the percentages of patients with substantial worsening, any worsening, worsening at endpoint, or improvement at endpoint in the HAM-D sleep disturbance factor in either insomnia subgroup. Treatments were well tolerated in most patients. No significant differences between treatments in the incidence of AEs suggestive of activation or sedation were seen in the insomnia subgroups. These data show no significant differences in acute treatment efficacy and tolerability across fluoxetine, sertraline, and paroxetine in major depressive disorder patients. Improvement in overall depression and in associated insomnia was achieved by most patients regardless of baseline insomnia. PMID- 11910259 TI - Sertraline and fluoxetine treatment of obsessive-compulsive disorder: results of a double-blind, 6-month treatment study. AB - The purpose of this study was to evaluate the comparative efficacy and tolerability of sertraline and fluoxetine in the treatment of obsessive compulsive disorder (OCD). Outpatients meeting DSM-IV criteria for OCD, with a Yale-Brown Obsessive-Compulsive (Y-BOCS) total score >or= 17, an NIMH Global Obsessive-Compulsive (NIMH-OC) scale score >or= 7, and a CGI-Severity score >or= 4 were randomized to 24 weeks of double-blind treatment with sertraline (N = 77) or fluoxetine (N = 73). Primary efficacy measures consisted of the Y-BOCS, the NIMH-OC scale, and the CGI-Severity (CGI-S) and Improvement (CGI-I) scales. Equivalent and significant (p < 0.001) improvement was found at week 24 in Y-BOCS and NIMH-OC scale scores for sertraline and fluoxetine. After 12 weeks, 49.2% of patients on sertraline were rated on the CGI-S scale as being mildly ill or not ill compared to 24.6% on fluoxetine (p < 0.01). A Cox analysis found patients on sertraline to have a statistically nonsignificant 42% greater likelihood of achieving a response by week 12 (CGI-I, much or very much improved; 95% CI, 0.85, 2.38; p = 0.18). Sertraline treatment also resulted in a higher proportion of remissions than fluoxetine (defined as a CGI-I 0.05). Time for the extrapolated DMR versus time log-linear plots to return to baseline was significantly different between fluoxetine (63.2 +/- 5.6 days) and both paroxetine (20.3 +/- 6.4 days) and sertraline (25.0 +/- 11.0 days) (p < 0.01), making the rank order (from longest to shortest) of time for CYP2D6 inhibition to dissipate: fluoxetine > sertraline >or= paroxetine. Differences between mean baseline DMR values and measured values obtained after drug discontinuation for each drug group became nonsignificant on discontinuation day 5 for both paroxetine and sertraline and on discontinuation day 42 for fluoxetine. These data define the time course of a persistent effect that fluoxetine, sertraline, and paroxetine have on CYP2D6 following drug discontinuation and should be considered when initiating therapy with a CYP2D6 substrate. PMID- 11910263 TI - Effect of fluoxetine and imipramine on the pharmacokinetics and tolerability of the antipsychotic quetiapine. AB - The effects of fluoxetine and imipramine on the pharmacokinetics and nonpsychiatric side effect profile of quetiapine fumarate were investigated in 26 patients with schizophrenia, schizoaffective disorder, or bipolar disorder in a multicenter, two-period, multiple-dose, open-label, randomized trial. Over a 1- to 2-week period, patients were titrated to a 300-mg twice-daily dose of quetiapine. Patients treated for at least 7 days at the target dose entered a combination therapy period, receiving fluoxetine (60 mg daily) or imipramine (75 mg twice daily) for 8 days. Key assessments included pharmacokinetic analysis of quetiapine, the Udvalg for kliniske undersogelser (UKU) Side Effect Rating Scale, and safety evaluations (e.g., adverse events, electrocardiograms, laboratory tests, and vital signs). Fluoxetine increased the quetiapine area under the plasma concentration time curve during a 12-hour interval (+12%), maximum plasma concentration during the dosing interval (C(ss)(max); +26%), and minimum plasma concentration at the end of the dosing interval (+8%), although it decreased oral clearance (-11%). The change in C(ss)(max) was statistically although not clinically significant. Imipramine did not affect the pharmacokinetics of quetiapine. Overall, scores on the UKU Side Effect Rating Scale improved during combination therapy with either agent, and no statistically significant deterioration was observed for any item. For safety assessments, the only clinically remarkable event was an imipramine-associated complete left bundle branch block in one patient. No unexpected side effects were reported. In conclusion, combination therapy with quetiapine and fluoxetine or imipramine had a minimal effect on quetiapine pharmacokinetics and was well tolerated. PMID- 11910264 TI - Placebo-controlled comparison of three dose-regimens of 5-hydroxytryptophan challenge test in healthy volunteers. AB - Single-dose administration of 5-hydroxytryptophan (5-HTP) is regularly used as a challenge test of the serotonergic system. The use of 5-HTP has been limited by an apparent small window between the occurrence of neuroendocrine endpoints and the occurrence of side effects. Therefore, many dosing strategies have been tried with and without concurrent administration of carbidopa, a peripheral inhibitor of the decarboxylation from 5-HTP to serotonin. The aim of the current study was to assess the relation between pharmacokinetics and pharmacodynamics of 5-HTP. Twelve healthy male volunteers were included in a placebo-controlled, randomized, four-way crossover, double-blind, single-dose investigation of oral 5-HTP with or without coadministration of carbidopa. The four dose regimens were placebo, 5-HTP 100 mg, 5-HTP 200 mg, and 5-HTP 100 mg with coadministration of carbidopa 100 mg and 50 mg at 3 hours before and 3 hours after the administration of 5-HTP, respectively. The last regimen resulted in a doubling of the elimination half life, an apparent clearance at least 14 times smaller, and a 15.4 times greater area under the curve compared with 5-HTP 100 mg without carbidopa. Furthermore, it was the only regimen to induce a significant change in cortisol and prolactin. It did not induce any change in subjective psychologic symptoms or cardiovascular parameters, but it was the only regimen to induce some nausea in three participants. The authors conclude that this regimen of 5-HTP 100 mg plus carbidopa is a relatively simple, effective, and tolerable challenge of the presynaptic serotonergic system. Further increase of the dose of 5-HTP might improve the size of the effect on endpoints as long as the tolerability remains good. PMID- 11910265 TI - Double-blind placebo-controlled pilot study of sertraline in military veterans with posttraumatic stress disorder. AB - The efficacy of sertraline in the treatment of civilian posttraumatic stress disorder (PTSD) has been established by two large placebo-controlled trials. The purpose of the current pilot study was to obtain preliminary evidence of the efficacy of sertraline in military veterans suffering from PTSD. Outpatient Israeli military veterans with a DSM-III-R diagnosis of PTSD were randomized to 10 weeks of double-blind treatment with sertraline (50-200 mg/day; N = 23, 83% male, mean age = 41 years) or placebo (N = 19, 95% male, mean age = 38 years). Efficacy was evaluated by the Clinician-Administered PTSD Scale (CAPS-2) and by Clinical Global Impression Scale-Severity (CGI-S) and -Improvement (CGI-I) ratings. Consensus responder criteria consisted of a 30% or greater reduction in the CAPS-2 total severity score and a CGI-I rating of "much" or "very much" improved. The baseline CAPS-2 total severity score was 94.3 +/- 12.9 for sertraline patients, which is notably higher than that reported for most studies of civilian PTSD. On an intent-to-treat endpoint analysis, sertraline showed a numeric but not statistically significant advantage compared with placebo on the CAPS-2 total severity and symptom cluster scores. In the study completer analysis, the mean CGI-I score was 2.4 +/- 0.3 for sertraline and 3.4 +/- 0.3 for placebo (t = 2.55, df = 30, p = 0.016), CGI-I responder rates were 53% for sertraline and 20% for placebo (chi2 = 3.62, df = 1, p = 0.057), and combined CGI I and CAPS-2 responder rates (>or=30% reduction in baseline CAPS-2 score) were 41% for sertraline and 20% for placebo (chi2 = 1.39, df = 1, p = 0.238). Sertraline treatment was well tolerated, with a 13% discontinuation rate as a result of adverse events. This pilot study suggests that sertraline may be an effective treatment in patients with predominantly combat-induced PTSD, although the effect size seems to be somewhat smaller than what has been reported in civilian PTSD studies. Adequately powered studies are needed to confirm these results and to assess whether continued treatment maintains or further improves response. PMID- 11910266 TI - Treatment predictors of extrapyramidal side effects in patients with tardive dyskinesia: results from Veterans Affairs Cooperative Study 394. AB - Predictors for the development of tardive dyskinesia (TD) have been studied extensively over the years, yet there are few studies of predictors of the course of TD after it has developed. Moreover, few studies have examined predictors of the course of other extrapyramidal side effects (EPS) in patients maintained on neuroleptics. The purpose of this study was to determine which modifiable variables are important in the prediction of EPS in patients with persistent TD over a period of as long as 2 years. One hundred fifty-eight patients enrolled in the Veterans Affairs Cooperative Study 394 were included in this study. A linear mixed-effects (LME) analysis to estimate the Abnormal Involuntary Movement Scale score (for TD severity), Simpson-Angus Scale (for parkinsonism severity), and Barnes Akathisia Scale at any given time after intake assessment was performed. The severity of each of the TD and EPS outcomes at any given visit was predicted by their respective baseline severity scores. Additional predictors of a favorable course of TD included lower doses of antipsychotic medications and use of anticholinergic medications. Other predictors of a favorable course of EPS included younger age and the use of atypical antipsychotic medication (for rigidity) and the use of anticholinergic medication (for tremor). These findings indicate that clinician-modifiable factors related to medication usage can influence the outcome of TD and EPS in patients with persistent TD. PMID- 11910267 TI - The effect of multiple doses of cimetidine on the steady-state pharmacokinetics of quetiapine in men with selected psychotic disorders. AB - Quetiapine fumarate (Seroquel) is an atypical antipsychotic agent approved for the treatment of psychosis. It is extensively metabolized by the CYP450 3A4 isozyme. The principal aim of the study was to investigate the effect of multiple doses of cimetidine, a nonspecific P450 inhibitor, on the steady-state pharmacokinetics of quetiapine. Thirteen patients (seven completers) with selected psychotic disorders received escalating doses of quetiapine from 25 to 150 mg three times daily on days 3 to 8 and were then maintained at 150 mg three times daily until day 19. Cimetidine (400 mg) was initiated on the afternoon of day 15 and administered three times daily with every dose of quetiapine thereafter. Quetiapine plasma concentrations were measured before and after cimetidine coadministration, and quetiapine pharmacokinetic parameters were calculated. Of the 13 men who entered the study, seven completed it. A slight increase in quetiapine plasma levels and reduction in oral clearance were observed after cimetidine coadministration. No serious adverse events were observed during quetiapine treatment. No clinically relevant alterations in quetiapine pharmacokinetics were observed after cimetidine coadministration in patients with psychotic disorders. PMID- 11910268 TI - The anxiolytic effect of the novel antipsychotic ziprasidone compared with diazepam in subjects anxious before dental surgery. AB - The novel atypical antipsychotic ziprasidone has a pharmacologic profile notable for potent agonism of serotonin (5-HT)1A receptors, antagonism at 5-HT1D receptors, and reuptake inhibition of norepinephrine. 5-HT1A receptor agonism, in particular, suggests anxiolytic activity, and ziprasidone has shown preliminary efficacy in treating the symptoms of anxiety associated with psychotic disorders. In this study, the anxiolytic efficacy of ziprasidone was evaluated in nonpsychotic subjects who were anxious before undergoing minor dental surgery. We compared a single oral dose of 20 mg ziprasidone (N = 30) with that of 10 mg diazepam (N = 30) and placebo (N = 30) in a randomized, parallel-group, double blind study. The peak anxiolytic effect of ziprasidone compared with that of placebo was similar to that of diazepam but had a later onset. At 3 hours postdose, the anxiolytic effect of ziprasidone was significantly greater than that of placebo (p < 0.05) and somewhat greater than that of diazepam. Diazepam showed a significantly greater anxiolytic effect than placebo at 1 hour (p < 0.05) but not at 3 hours. The sedative effect of ziprasidone was never greater than that of placebo, whereas that of diazepam was significantly greater than that of placebo 1 to 1.5 hours postdose. Ziprasidone was generally well tolerated. Only one patient reported treatment-related adverse events (nausea and vomiting) and, unlike diazepam, ziprasidone did not cause reductions in blood pressure. Dystonia, extrapyramidal syndrome, akathisia, and postural hypotension were not seen with ziprasidone. Thus, ziprasidone may possess anxiolytic effects in addition to its antipsychotic properties. PMID- 11910269 TI - Paroxetine increases steady-state concentrations of (R)-methadone in CYP2D6 extensive but not poor metabolizers. AB - Steady-state blood concentrations of (R)- methadone (i.e., the active form), (S) methadone, and (R,S)-methadone were measured before and after introduction of paroxetine 20 mg/day during a mean period of 12 days in 10 addict patients in methadone maintenance treatment. Eight patients were genotyped as CYP2D6 homozygous extensive metabolizers (EMs) and two patients as poor metabolizers (PMs). Paroxetine significantly increased concentrations of both enantiomers of methadone in the whole group (mean increase for (R)-methadone +/- SD, 26 +/- 32%; range, -14% to +83%, p = 0.032; for (S)-methadone, 49 +/- 51%; range, -29% to +137%, p = 0.028; for (R,S)-methadone, 35 +/- 41%; range, -20% to +112%, p = 0.032) and in the group of eight EMs (mean increase, 32%, p = 0.036; 53%, p = 0.028; and 42%, p = 0.036, for (R)-methadone, (S)-methadone, and (R,S)-methadone, respectively). On the other hand, in the two PMs, (S)-methadone but not (R) methadone concentrations were increased by paroxetine (mean increases of 36% and 3%, respectively). Paroxetine is a strong CYP2D6 inhibitor, and these results confirm previous studies showing an involvement of CYP2D6 in methadone metabolism with a stereoselectivity toward the (R)-enantiomer. Because paroxetine is a mild inhibitor of CYP1A2, CYP2C9, CYP2C19, and CYP3A4, increase of (S)-methadone concentrations in both EMs and PMs could be mediated by inhibition of any of these isozymes. PMID- 11910270 TI - Double-blind, placebo-controlled study of single-dose metergoline in depressed patients with seasonal affective disorder. AB - A role for serotonin in season affective disorder (SAD) has been explored with a variety of serotonergic pharmacologic agents. The authors initially hypothesized that metergoline, a nonspecific serotonin antagonist, would exacerbate depressive symptoms. In a small, open-label pilot study, the authors observed the opposite effect. They decided to follow up on this finding with this formal study. The study followed a double-blind, randomized cross-over design. Sixteen untreated, depressed patients with SAD received single oral doses of metergoline 8 mg and of placebo, spaced 1 week apart. Fourteen patients were restudied after 2 weeks of light treatment. Depression ratings using the Structured Interview Guide for the Hamilton Depression Rating Scale-Seasonal Affective Disorder Version were performed at baseline and at 3 and 6 days after each intervention. These data were analyzed by baseline-corrected repeated measures with analysis of variance. In the off-lights condition, severity of depression was diminished after metergoline compared with placebo administration (p = 0.001). Patient daily self ratings suggested that the peak effect occurred 2 to 4 days after study drug administration. In contrast, after 2 weeks of treatment with bright artificial light, metergoline did not demonstrate a significant effect on mood. These data suggest that single doses of metergoline may have antidepressant effects that last several days. Possible mechanisms include 5-hydroxytryptamine(2) receptor downregulation and dopamine agonism. PMID- 11910271 TI - Acute stress-induced seizures and loss of consciousness in a ten-year-old boy with cutaneous mastocytosis. PMID- 11910272 TI - Effects of oral procyclidine administration on cognitive functions in healthy subjects: implications for schizophrenia. PMID- 11910273 TI - Donepezil management of schizophrenia with associated dementia. PMID- 11910275 TI - Pathologic laughter associated with paroxetine treatment. PMID- 11910274 TI - Amitriptyline and paroxetine: effects upon peripheral nervous system (PNS). PMID- 11910276 TI - Conservative management with naltrexone of an iatrogenic methadone overdose in an opiate-naive patient. PMID- 11910277 TI - Possible duplicate publication. PMID- 11910279 TI - Salt consumption, reactive oxygen species and cardiovascular ageing: a hypothetical link. AB - Opinions about the impact of dietary salt on blood pressure have dominated the debate regarding 'salt sensitivity', which can be broadly defined as the blood pressure response to changes in sodium intake among individuals in a population. However, the larger question is whether salt consumption exerts significant biological effects independent of changes in blood pressure. Provisional answers to this question are reviewed based on newly discovered links between sodium metabolism and the generation of reactive oxygen species. These links suggest that, in a subset of the general population, salt consumption is a determinant in cardiovascular ageing. PMID- 11910280 TI - Salt-sensitivity, hypertension and cardiovascular ageing: broadening our view without missing the point. PMID- 11910281 TI - Hypertension and the risk of cancer: is there new evidence? PMID- 11910282 TI - Relationship between size at birth and hypertension: problems and perspectives. PMID- 11910283 TI - Self blood pressure monitoring at home by wrist devices: a reliable approach? PMID- 11910284 TI - Reversed white-coat hypertension: definition, mechanisms and prognostic implications. PMID- 11910285 TI - The drive to identify genetic factors influencing left ventricular mass responses to antihypertensive treatment. PMID- 11910286 TI - Endothelins and venous tone in DOCA-salt hypertension. PMID- 11910287 TI - Right enzyme? Wrong place? PMID- 11910288 TI - Monocytes/macrophages in hypertension. PMID- 11910289 TI - Angiotensin and epidermal growth factor receptor cross talk goes up and down. PMID- 11910290 TI - Neutral endopeptidase inhibition: the potential of a new therapeutic approach in cardiovascular disease evolves. PMID- 11910291 TI - NKCC2: a drug target in hypertension. PMID- 11910292 TI - Dual angiotensin II blockade: a promise of enhanced renal protection? PMID- 11910293 TI - Left ventricular hypertrophy: a new approach for fibrosis inhibition. PMID- 11910294 TI - The ABCD of anti-hypertensive therapy? PMID- 11910295 TI - Does hypertension increase mortality risk from lung cancer? A prospective cohort study on smoking, hypertension and lung cancer risk among Korean men. AB - OBJECTIVE: To examine the effects of hypertension on lung cancer prospectively and to determine the interactive effect of hypertension and smoking on lung cancer risk. DESIGN: A prospective cohort study. PARTICIPANTS: The cohort comprised 452,645 Korean men, aged 35-64 years, who received health insurance from the Korea Medical Insurance Corporation and who had biennial medical evaluations in 1992 and 1994. METHODS: Multivariate Cox proportional hazard models were tested, controlling for age, smoking status, exercise, body mass index, alcohol use, diabetes and serum cholesterol concentration. MAIN OUTCOME MEASURE: Deaths from lung cancer. RESULTS: At baseline, 261 080 persons (58.3%) were identified as current cigarette smokers. Between 1995 and 1999, 883 deaths from lung cancer (44.8/100,000 person-years) occurred. An initial finding indicated that hypertension increased the mortality risk of lung cancer [risk ratio (RR) 1.3, 95% confidence interval (CI) 1.1-1.5]. However, after stratification for smoking status, the risk ratio was increased only for current smokers (RR 1.4, 95% CI 1.2-1.6). When the interaction term was included in the multivariate model, there was a significant interactive effect of hypertension with current smoking (RR 1.8, 95% CI 1.0-3.1) on the risk of death from lung cancer, whereas the effect of hypertension itself did not attain significance. CONCLUSION: Hypertension was not an independent risk factor in lung cancer related deaths, but it increased the modest risk of lung cancer death among current smokers. PMID- 11910296 TI - Relationship between size at birth and hypertension in a genetically homogeneous population of high birth weight. AB - OBJECTIVE: To investigate the association between birth size and hypertension within a genetically homogeneous population of high birth weight. DESIGN: Cohort study with retrospectively collected data on size at birth. SUBJECTS AND SETTING: The study included 4601 men and women born 1914-1935 in Reykjavik, Iceland, who participated in the Reykjavik Study of the Icelandic Heart Association. MAIN OUTCOME MEASURES: Birth size measurements, adult blood pressure (BP) and body mass index (BMI), and family history of hypertension. RESULTS: Birth weight was inversely related to hypertension in adulthood in women (P for trend < 0.001). The relationship was of borderline significance in men (P for trend = 0.051). A low ponderal index was significantly associated with high BP in women (P for trend = 0.025) but not men (P > 0.05). For women with an adult BMI > 26 kg/m2, the odds ratio for hypertension for those born weighing < 3.45 kg was 2.1 [95% confidence interval, 1.3-3.3, compared with women born weighing > 3.75 kg. The association was only significant in women without a family history of hypertension. CONCLUSIONS: An inverse association between size at birth and adult hypertension was seen in a population of greater birth size than has previously been investigated. The relation was strongest among women born small who were overweight in adulthood, and for those without a family history of hypertension. The results support the hypothesis that the association between birth weight and hypertension is not of genetic origin only. The large birth size of Icelanders might be protective and partly explain the lower mean systolic blood pressure in Iceland than in related nations. PMID- 11910297 TI - Accuracy and reliability of wrist-cuff devices for self-measurement of blood pressure. AB - OBJECT: Self-measurement of blood pressure (BP) might offer some advantages in diagnosis and therapeutic evaluation and in patient management of hypertension. Recently, wrist-cuff devices for self-measurement of BP have gained more than one third of the world market share. In the present study, we validated wrist-cuff devices and compared the results between wrist- and arm-cuff devices. The factors affecting the accuracy of wrist-cuff devices were also studied. METHOD: The research group to assess the validity of automated blood pressure measuring device consisted of 13 institutes in Japan, which validated two wrist-cuff devices (WC-1 and WC-2) and two arm-cuff devices (AC-1 and AC-2). They used a crossover method, where the comparison was done between auscultation, by two observers by means of a double stethoscope on one arm and the device on the opposite arm or wrist. RESULTS: There was good inter-observer agreement for the auscultation method in each institute (systolic blood pressure (SBP), -0.1 +/- 2.8 mmHg; diastolic blood pressure (DBP), -0.1 +/- 2.6 mmHg, n = 498). The mean difference between auscultation and the device was minimal both in arm-cuff devices (mean difference for AC-1, 2.2/1.9 mmHg, n = 97 and for AC-2, 5.1/2.9 mmHg, n = 136, SBP/DBP) and wrist-cuff devices (mean difference for WC-1, 2.1/1.2 mmHg, n = 173 mmHg and for WC-2, -2.3/-5.6 mmHg, n = 92). The standard deviation of the difference (SDD) in wrist-cuff devices, however (SDD for WC-1, 9.7/7.3 mmHg and for WC-2, 10.2/8.6 mmHg), was larger than that of the arm-cuff devices (SDD for AC-1, 5.6/6.6 mmHg and for AC-2, 6.3/5.1 mmHg). Grading of AC-1 and AC-2 based on criteria of British Hypertension Society was A/A and B/A, respectively, while that of WC-1 and WC-2 was C/B and D/B, respectively. Using the same validation protocol, the results of validation for one device were divergent in each institute. In wrist-cuff devices, the BP value obtained in palmar flexion was significantly higher and that obtained in palmar dorsiflexion was significantly lower than that in palmar extension. In some cases, finger plethysmogram did not disappear during maximum inflation of the wrist-cuff (congruent with 250 mmHg), even in palmar extension and especially in palmar flexion, suggesting incomplete obstruction of radial and/or ulnar arteries during inflation. CONCLUSION: The results suggest that wrist-cuff devices in the present form are inadequate for self-measurement of blood pressure and, thus, are inadequate for general use or clinical and practical use. However, there is much possibility in wrist-cuff device and the accuracy and reliability of wrist-cuff device are warranted by an improvement of technology. PMID- 11910298 TI - 'Reverse white-coat hypertension' in older hypertensives. AB - OBJECTIVES: The role of ambulatory blood pressure monitoring (ABPM) in the elderly is unclear. This study has examined differences between clinic and ambulatory blood pressure (BP) in a large cohort of older hypertensives, with particular respect to the factors influencing the direction and magnitude of this difference. DESIGN: The Second Australian National Blood Pressure Study (ANBP2) is a general practice-based randomized-outcome trial in 6083 older hypertensives treated with an angiotensin-converting enzyme (ACE) inhibitor or diuretic-based regimen. Before starting treatment a subset of 713 patients (age range 65-83 years) had a 'successful' 26-hour ambulatory blood pressure recording with a SpaceLabs 90207 recorder. RESULTS: Average clinic BP (+/- SD) was 167 +/- 12/90 +/- 8 mmHg. Average daytime ambulatory BP was 157 +/- 15*/89 +/- 10* mmHg and night ambulatory BP was 137 +/- 16+/74 +/- 10+ mmHg (different from clinic BP: *P < 0.01;from daytime ambulatory BP: +P < 0.001). Twenty-one to 45% of all patients had higher daytime systolic or diastolic ambulatory BP than clinic readings, with smoking, previous treatment for hypertension and lower clinic BP being the main predictors of this 'reverse white-coat effect'. CONCLUSIONS: Although mean daytime ambulatory blood pressures were lower than clinic readings in this large cohort of untreated older hypertensives, a substantial proportion showed the reverse of the so-called 'white-coat effect'. These findings identify the important role for ABPM in the elderly, not only for avoiding overtreatment in those with typical 'white-coat hypertension' but also for ensuring adequate treatment is given to those with the reverse of this phenomenon. PMID- 11910299 TI - The influence of zygosity status on blood pressure and on lipid profiles in male and female twins. AB - OBJECTIVE: To study the influence of zygosity on blood pressure and serum lipid concentrations among male and female twins. SETTING: Department of Endocrinology, Odense University Hospital, Denmark. PARTICIPANTS: A total of 125 monozygotic and 178 dizygotic twin pairs aged 55-74 years of age, ascertained from The Danish Twin Register. DESIGN: Population-based cross-sectional study. MAIN OUTCOME MEASURES: Blood pressure and serum lipid concentrations. RESULTS: The prevalence of hypertriglyceridemia and hypercholesterolemia were higher among monozygotic compared with dizygotic twins, whereas the prevalence of hypertension was similar. The level of triglycerides [0.28 (0.44) versus 0.18 (0.41), P = 0.01] and total cholesterol [1.82 (0.17) versus 1.78 (0.19), P = 0.03] were significantly higher in monozygotic compared with dizygotic twins. Systolic blood pressure was non-significantly higher among monozygotic twins (136.8 (21.3) versus 134.1 (19.6), P = 0.10). When comparing monozygotic and dizygotic twins within each sex group, the difference in triglyceride level was only apparent among male twins and the differences in systolic blood pressure and total cholesterol were only seen among female twins. Birth weight as determined in a subgroup of the population was similar in monozygotic and dizygotic twins. CONCLUSIONS: Zygosity status per se influences fasting serum triglycerides and total-cholesterol and to some extent systolic blood pressure in twins, supporting an influence of an intrauterine component on lipid profiles. The influence is independent of birth weight and seems to be sex-specific. PMID- 11910300 TI - Polymorphisms of genes encoding components of the sympathetic nervous system but not the renin-angiotensin system as risk factors for orthostatic hypotension. AB - OBJECTIVE: The genetic background of orthostatic hypotension, an important risk factor for future cardiovascular morbidity and mortality, was investigated. DESIGN AND METHODS: The study subjects comprised 415 community-dwelling individuals, who were free from any cardiovascular complications, aged 50 years or older (mean age 70.5 +/- 9 years). Basal systolic blood pressure (SBP) was measured twice in supine posture after resting for more than 10 min. The orthostatic change in SBP was determined at 1 min and 3 min after standing up. The maximum change in SBP after standing was determined. Orthostatic hypotension was defined as a decline in SBP greater than 20 mmHg. The polymorphisms of genes encoding components of the renin-angiotensin system and sympathetic nervous system, which play pivotal roles in postural change in blood pressure regulation, were determined. RESULTS: There were no significant associations between the maximum change in SBP, the prevalence of orthostatic hypotension and gene polymorphisms of angiotensin-converting enzyme I/D, angiotensinogen M235T and angiotensin II type 1 receptor A1166C. On the contrary, polymorphism of the Gs protein alpha-subunit (GNAS1) T131C was significantly associated with the maximum change in SBP after standing [1.9 +/- 16 versus -3.6 +/- 16 mmHg (TT + TC versus CC), P = 0.008]. The prevalence of orthostatic hypotension was significantly different among GNAS1 genotypes (chi squared = 10.12, P = 0.011) and G-protein beta 3 subunit (GNB3) genotypes (chi squared = 6.12, P = 0.020). Multiple logistic regression analysis showed that both GNAS1 CC genotype [odds ratio (OR) = 2.79, 95% confidence interval (CI) 1.35-5.79, P = 0.006] and GNB3 C allele (OR = 1.78, 95% CI 1.06-3.00, P = 0.030) were independent risks for orthostatic hypotension. CONCLUSIONS: These findings indicate that genes encoding sympathetic nervous components could be involved in the predisposition for orthostatic hypotension. PMID- 11910301 TI - Polymorphisms in the angiotensinogen and angiotensin II type 1 receptor gene are related to change in left ventricular mass during antihypertensive treatment: results from the Swedish Irbesartan Left Ventricular Hypertrophy Investigation versus Atenolol (SILVHIA) trial. AB - BACKGROUND: Our aim was to determine if gene polymorphisms in the renin angiotensin-aldosterone system (RAAS) were related to the degree of change in left ventricular hypertrophy (LVH) during antihypertensive treatment. METHODS AND RESULTS: Patients with essential hypertension and echocardiographically diagnosed LVH were included in a double-blind study to receive treatment with either the angiotensin II type 1 receptor (AT1-receptor) antagonist irbesartan (n = 41), or the beta-1 adrenergic receptor blocker atenolol (n = 43) as monotherapy for 3 months. The angiotensinogen T174M and M235T, the angiotensin-converting enzyme I/D, the AT1-receptor A1166C and the aldosterone synthase (CYP11B2) -344 C/T polymorphisms were analysed and related to the change in left ventricular mass (LVM). Patients with the angiotensinogen 174 TM genotype treated with irbesartan responded with the greatest reduction in LVM (-23 +/- 31SD g/m2 for TM and +0.5 +/- 18 g/m2 for TT, P = 0.005), independent of blood pressure reduction. Both the angiotensinogen 235 T-allele (P = 0.02) and the AT1-receptor 1166 AC genotype responded with the greatest reduction in LVM when treated with irbesartan (-0.1 +/- 19 g/m2 for AA and -18 +/- 30 g/m2 for AC, P = 0.02), independent of blood pressure reduction. These polymorphisms were not associated with the change in LVM during treatment with atenolol. DISCUSSION: The angiotensinogen T174M and M235T and the AT1-receptor A1166C polymorphisms were related to the change in LVH during antihypertensive treatment with an AT1-receptor antagonist; of these angiotensinogen T174M was the most powerful. This highlights the role of the RAAS for left ventricular hypertrophy and the potential of pharmacogenetics as a tool for guidance of antihypertensive therapy. PMID- 11910302 TI - Endothelin receptor function in mesenteric veins from deoxycorticosterone acetate salt-hypertensive rats. AB - OBJECTIVES: To identify the receptors by which endothelin-1 (ET-1) increases venomotor tone in hypertension. METHODS: Vascular reactivity to ET-1 and the selective endothelin receptor subtype B (ET(B)) agonist, sarafotoxin 6c (S6c), was studied in mesenteric blood vessels from deoxycorticosterone acetate (DOCA salt) hypertensive and normotensive control rats. The diameter of small (< or = 280 microm) mesenteric arteries and veins was monitored in vitro using computer assisted video microscopy. Contractions of mesenteric arteries (< or= 250 microm diameter) were also studied, using a myograph. ET-1 mRNA levels were measured in mesenteric arteries and veins using real-time RT-PCR techniques. RESULTS: ET-1 induced contractions were reduced in arteries of DOCA-salt hypertensive rats compared with those of normotensive control rats; S6c produced negligible contractions in arteries from both groups. ET-1 concentration-responses curves in arteries measured using video microscopy or a myograph were similar. ET-1 and S6c caused veins to contract, and there were no differences between responses to these agonists in tissues from DOCA-salt hypertensive rats or normotensive control rats. Studies using ET(A) and ET(B) receptor antagonists indicated that ET-1-induced venoconstriction was mediated by ET(A) receptors. Potassium chloride concentration-response curves were similar in arteries and veins from normotensive control rats and DOCA-salt hypertensive rats. ET-1 mRNA levels in DOCA-salt hypertensive rat arteries or veins were not different from those in normotensive control rat arteries and veins. CONCLUSIONS: These data indicate that ET-1 reactivity is maintained in mesenteric veins, but not arteries, in DOCA salt hypertension. Therefore, the sustained increase in venomotor tone mediated by ET(A) receptors that is known to occur in vivo in DOCA-salt hypertensive rats is not caused by direct venoconstriction. PMID- 11910303 TI - Increased free radical production in hypertension due to increased expression of the NADPH oxidase subunit p22(phox) in lymphoblast cell lines. AB - OBJECTIVES: To confirm increased production of reactive oxygen species (ROS) in hypertension, to demonstrate the source of ROS and to analyse NADPH oxidase subcomponent expression in hypertension. DESIGN: A lymphoblast model was used, as this has previously been used in the study of hypertension and of NADPH oxidase. Chemiluminescence (CL) was chosen to assay ROS production, as it is simple and sensitive. METHODS: Lymphocytes from 12 hypertensive patients (HT), and 12 age- and sex-matched normotensive (NT) subjects, were immortalized. Luminol, isoluminol and Cypridina luciferin analogue (CLA) CL were used to assay ROS production. NADPH oxidase subunits were measured by Western blot analysis. RESULTS: Stimulation with 50 micromol/l arachidonic acid (AA) resulted in increased ROS production in HT cell lines with luminol, CLA and isoluminol CL. Stimulation with 500 nmol/l 12-O-tetradecanoylphorbol-13-acetate (TPA) produced a detectable increase in HT ROS production with luminol and with CLA, whereas there was no significant difference with isoluminol. The ROS production was abolished by diphenyleneiodonium chloride (DPI) but not by rotenone, indicating that a non mitochondrial flavoprotein such as NADPH oxidase is the source of ROS. Analysis of NADPH oxidase subcomponents revealed an increase in p22(phox) in HT subjects. CONCLUSIONS: We have shown there is increased ROS production in lymphoblasts derived from hypertensive subjects, probably originating from NADPH oxidase. As the ROS production persists in transformed cells, this suggests a genetic predisposition to increased ROS production. Increased expression of p22(phox) in HT lymphoblasts may account for some of the increased ROS. PMID- 11910304 TI - Identification of mechanically induced genes in human monocytic cells by DNA microarrays. AB - BACKGROUND: Hypertension is a risk factor for coronary heart disease. Macrophages are critically involved in both atherogenesis and plaque instability. Although macrophages may be subjected to excess mechanical stress in these diseases, the way in which biomechanical forces affect macrophage function remains incompletely defined. OBJECTIVE: To investigate the molecular response to mechanical force in macrophages. DESIGN AND METHODS: We used a DNA microarray with 1056 genes to describe the transcriptional profile of mechanically induced genes in human monocytic THP-1 cells. Mechanical deformation was applied to a thin and transparent membrane on which cells were cultured. After THP-1 cells were pre incubated in the presence of phorbol 12-myristate 13-acetate (0.2 micromol/l) for 24 h, THP-1 cells attached to the membrane were subjected to biaxial mechanical strain. Interleukin-8 concentrations were determined using an enzyme-linked immunosorbent assay. RESULTS: In DNA microarray analysis, cyclic mechanical strain at 1 Hz induced only three genes more than 2.5-fold at 3 and 6 h in THP-1 cells: prostate apoptosis response-4 (3.0-fold at 3 h, 6.7-fold at 6 h), interleukin-8 (4.3-fold at 6 h) and the immediate-early response gene, IEX-1 (2.6 fold at 6 h). Real-time reverse transcriptase polymerase chain reaction analysis confirmed the amplitude-dependent induction of these three genes. In addition, mechanical strain increased interleukin-8 protein expression. CONCLUSION: The present study demonstrates that human monocytic cells respond to mechanical deformation with induction of immediate-early and inflammatory genes. These findings suggest that mechanical stress in vivo, such as that associated with hypertension, may play an important part in atherogenesis and instability of coronary artery plaques, through biomechanical effects on vascular macrophages. PMID- 11910305 TI - Functional cross-talk between angiotensin II and epidermal growth factor receptors in NIH3T3 fibroblasts. AB - BACKGROUND: The main angiotensin (Ang) II subtype receptors (AT1R and AT2R) are involved in cellular growth processes and exert functionally antagonistic effects. OBJECTIVE: To characterize the mechanisms by which Ang II receptors influence growth, by investigating the interactions between Ang II subtype receptors and epidermal growth factor (EGF) receptors on mitogen-activated protein kinase (MAPK) pathway activation. DESIGN AND METHODS: The experiments were performed using a mouse fibroblast cell line, NIH3T3, by transient co transfection with rat AT1R or AT2R expression vectors, or both. Extracellular signal-regulated kinase (ERK)1/2 phosphorylation was analysed by western blot and the ERK activity was evaluated using PathDetect, an in-vivo signal transduction pathway trans-reporting system. Selective Ang II receptor antagonists (losartan for AT1R and PD123319 for AT2R) were used to investigate the contributions of each receptor to the response observed. RESULTS: Our data show that, in this cellular model, both Ang II receptors phosphorylate ERK1/2. However, in the cells expressing AT1R, the EGF-induced MAPK pathway was enhanced in the presence of Ang II in a synergistic fashion. In contrast, a reduction of EGF-induced MAPK activation was observed in the cells expressing AT2R. In cells expressing both Ang II subtype receptors, Ang II promoted an enhancement of EGF-induced MAPK activation. However, in the presence of the AT1R antagonist, losartan, the effect of EGF was reduced. CONCLUSION: These data indicate the existence of an opposite cross-talk of AT1R and AT2R with EGF receptors, and suggest a complex functional interaction between these pathways in the regulation of cellular growth processes. PMID- 11910306 TI - Heterogeneity of autonomic regulation in hypertension and neurovascular contact. AB - OBJECTIVE: Brainstem neurovascular contact (NVC) may interfere with central autonomic regulation and contribute to essential hypertension. We have previously shown that patients with autosomal-dominant hypertension, brachydactyly, and NVC feature extreme phenylephrine hypersensitivity due to impaired baroreflex buffering. We tested the hypothesis that similar abnormalities are present in patients with essential hypertension who have NVC. METHODS: Six patients with NVC and essential hypertension and five patients with NVC and monogenic hypertension and brachydactyly were studied. Responses to incremental phenylephrine doses were assessed before and during ganglionic blockade with trimethaphan. RESULTS: Supine blood pressure was 172 +/- 8.8/89 +/- 6.1 mmHg before ganglionic blockade. Blood pressure decreased 47 +/- 5/18 +/- 3 mmHg with trimethaphan (16 +/- 4.4/4 +/- 4.0 mmHg in autosomal-dominant hypertension, P < 0.05). Before ganglionic blockade, 25 microg phenylephrine increased systolic blood pressure 17 +/- 4 mmHg in patients with essential hypertension and 30 +/- 3 mmHg in patients with autosomal dominant hypertension (P < 0.05). During ganglionic blockade, the same dose increased systolic blood pressure 32 +/- 1 and 33 +/- 4 mmHg in patients with essential and with autosomal-dominant hypertension, respectively (NS). CONCLUSIONS: Phenylephrine hypersensitivity due to baroreflex dysfunction is uncommon in patients with essential hypertension and NVC. This finding may suggest that the effect of NVC on autonomic regulation is heterogeneous. An alternative explanation is that radiological NVC is not necessarily functionally relevant. PMID- 11910307 TI - Combined inhibition of neutral endopeptidase with angiotensin converting enzyme or endothelin converting enzyme in experimental diabetes. AB - OBJECTIVE: The effects of combined inhibition of neutral endopeptidase (NEP) with either angiotensin-converting enzyme (ACE), or endothelin-converting enzyme (ECE) on blood pressure, urinary albumin excretion and heart weight were explored in experimental diabetes. DESIGN: Streptozotocin-induced diabetic Sprague-Dawley rats were treated with vehicle, the NEP/ACE inhibitor S 21402, the NEP/ECE inhibitor CGS 26303, the NEP inhibitor SCH 42495, the ACE inhibitor captopril or the endothelin receptor antagonist bosentan for 4 weeks. METHODS: Blood pressure was measured by tail-cuff method and radiotelemetry. Albuminuria, plasma renin activity and plasma atrial natriuretic peptide (ANP) were determined by radioimmunoassay. NEP binding was assessed by in vitro quantitative autoradiography. Metabolic and biochemistry parameters including food intake, 24 h urine volume, plasma glucose, glycated hemoglobin, glomerular filtration rate (GFR) and urinary sodium excretion were also determined. RESULTS: Mean blood pressure over the 4-week study period after commencement of treatment was reduced to a similar extent by a range of treatments including the ACE inhibitor, NEP/ACE inhibitor, endothelin receptor antagonist, NEP/ECE inhibitor, but not the NEP inhibitor, compared with vehicle-treated diabetic rats. Heart to body weight ratio in diabetic rats was only reduced by the NEP/ACE and the NEP/ECE inhibitor. Increased albuminuria in diabetic rats (1.1 times/divided by 1.2 mg/day) was reduced by the NEP/ACE (0.6 times/divided by 1.2 mg/day) and the NEP/ECE inhibitors (0.4 times/divided by 1.2 mg/day). Renal NEP was reduced by the NEP/ACE inhibitor (35 +/- 4%) or NEP/ECE inhibitor (38 +/- 4%) as well as by the pure NEP inhibitor (27 +/- 4%) compared with the untreated diabetic group. Other abnormal metabolic and biochemical parameters in diabetic rats were not influenced by any drug treatment. CONCLUSIONS: Combined inhibition of NEP/ACE or NEP/ECE confers beneficial effects on blood pressure, albuminuria and heart to body weight ratio in experimental diabetes. PMID- 11910308 TI - Losartan titration versus diuretic combination in type 2 diabetic patients. AB - METHODS: We compared the effects of Losartan dose titration to 100 mg versus the addition of 12.5 mg of hydrochlorothiazide, in 90 type 2 diabetic patients with microalbuminuria and blood pressure > 130/85 mmHg, receiving losartan 50 mg as initial treatment during 4 weeks. RESULTS: With the first dose of losartan, systolic (SBP) and diastolic blood pressure (DBP) decreased from 154.5 (152.1 157.5) to 144.4 (141.3-147.5) mmHg (P < 0.001) and from 91.1 (89.4-92.8) to 84.6 (82.8-86.4) mmHg (P < 0.001), with 20 patients attaining the expected goal blood pressure (< 130/85 mmHg); albuminuria decreased from 109.8 (90.5-133.3) to 83.5 (63.6-109.5) mg per 24 h (P = 0.006). Patients not attaining the target blood pressure were randomly allocated to titration or to the combination arm. After an additional 4 weeks, patients titrated exhibited a fall in SBP and DBP from 157.1 (152.7-161.5) to 142.1 (136.4-147.8) mmHg (P < 0.001) and from 92.4 (89.5-95.3) to 83.6 (81.1-86.1) mmHg (P < 0.001); albuminuria decreased from 136.3 (97.8 189.9) to 99.7 (69.3-143.4) mg per 24 h (P = 0.002). In the combination arm, there were similar reductions in SBP and DBP from 155.3 (151.5-159.1) to 139.1 (132.1-146.1) mmHg (P < 0.001) and from 92.1 (89.3-94.9) to 80.9 (77.4-84.4) mmHg (P < 0.001); while albuminuria fell from 107.7 (82.2-141.0) to 64.2 (45.9-89.9) mg per 24 h (P = 0.001). CONCLUSIONS: Losartan 50 mg was effective in reducing blood pressure and albuminuria in type 2 diabetic patients. When the blood pressure target was not reached, the two strategies tested seem to contribute similarly to further reductions in blood pressure and albuminuria. PMID- 11910309 TI - Renal Na-K-Cl cotransporter NKCC2 in Dahl salt-sensitive rats. AB - BACKGROUND: Dahl salt-sensitive (DS) rats are characterized by enhanced NaCl reabsorption in the loop of Henle, but the responsible ion transport protein is unknown. OBJECTIVE: To investigate renal Na-K-Cl cotransporter NKCC2 function and expression in DS rats under a low-salt diet. METHODS: NKCC2 functioning was assessed in vitro by measuring bumetanide-sensitive rubidium uptake and cytosolic chloride concentration in isolated medullary thick ascending limb (mTAL) tubules, and in vivo by measuring the salidiuretic action of orally given bumetanide. NKCC2 expression was assessed by Western blot analysis of outer medullary proteins using T4 monoclonal antibody. RESULTS: mTAL tubules from DS rats exhibited significantly higher bumetanide-sensitive rubidium uptake (85.1 +/- 4.8 versus 66.2 +/- 4.4 nmol/min per mg protein in DS and DR, (Dahl salt-resistant) rats, respectively; P = 0.011) and significantly higher cytosolic chloride (32.8 +/- 1.7 versus 25.0 +/- 1.5 mmol/l in DS and DR rats, respectively). Moreover, DS rats showed a significantly higher (P < 0.001) natriuretic response to bumetanide (1.13 +/- 0.05 versus 0.64 +/- 0.09 mmole/3 h in DS and DR rats, respectively). Finally, Western blot analysis revealed less NKCC2 expression in DS rats. CONCLUSIONS: We conclude that DS rats have increased renal NKCC2 activity, thus explaining, at least in part, their genetic renal inability to excrete sodium. Moreover, DS rats have a decreased renal NKCC2 expression, which can be a compensatory phenomenon against NKCC2 hyperactivity. PMID- 11910310 TI - Dissociation between blood pressure reduction and fall in proteinuria in primary renal disease: a randomized double-blind trial. AB - OBJECTIVE: Guidelines recommend lower threshold and goal blood pressure (BP) for patients with proteinuria. BP reduction could be accompanied by a different fall in proteinuria depending of the antihypertensive drug. The objective was to compare proteinuria reduction when BP is lowered to the same level with different drugs. DESIGN: Prospective, randomized, double-blind, controlled trial. SETTING: 12 Spanish centres. PATIENTS: A total of 119 patients with primary renal disease, blood pressure > 130/85 mmHg, proteinuria > 1 g/day, and creatinine clearance > or = 50 ml/min. INTERVENTION: After a 4-week run-in placebo period, patients were randomized to: atenolol 50 mg/day; trandolapril 2 mg/day; verapamil 240 mg/day or verapamil 180 + trandolapril 2 mg/day combination; forced double-dose titration was carried out at the 4th week. Treatment duration was 6 months. OUTCOME MEASURES: Changes in BP, 24 h proteinuria, serum albumin and calcium. RESULTS: BP was significantly reduced with the four treatments [SBP/DBP (mmHg]: atenolol 12.2/9.9; trandolapril 12.9/9.3; verapamil 8.2/7.9 and verapamil + trandolapril 13.6/11.3) without differences between them. A significant fall in proteinuria was seen in the trandolapril, 40.2% [95% confidence interval (CI) 24.3-56.2%], and verapamil + trandolapril groups, 48.5% (95% CI, 31.7-64.3%) accompanied with increases in serum albumin (trandolapril: from 3.86 +/- 0.64 to 4.03 +/- 0.67 g/dl; verapamil + trandolapril: from 4.15 +/- 0.58 to 4.40 +/- 0.51 g/dl). CONCLUSIONS: In patients with proteinuric primary renal disease, adequate dose titration of antihypertensive drugs may provide a substantial BP reduction. Only angiotensin-converting enzyme inhibitor (trandolapril) treatment, alone or better combined with verapamil, reduces proteinuria and increases serum albumin. PMID- 11910311 TI - Reduction of proteinuria; combined effects of receptor blockade and low dose angiotensin-converting enzyme inhibition. AB - OBJECTIVE: Angiotensin-converting enzyme inhibitors (ACEI) show an antiproteinuric and thus nephroprotective effect in patients suffering from glomerulonephritis. Angiotensin II-receptor-antagonists (AT1RA) are also efficacious in reducing proteinuria. The study was performed to investigate the antiproteinuric effect of AT1RA candesartan in patients diagnosed with chronic glomerulonephritis by biopsy, and who were already being treated with an ACEI. METHODS: A total of 12 patients with a persistent proteinuria of at least 1 g/day who were already being treated with an ACEI for more than 3 months were included. The study was performed using a double-blind, placebo-controlled and randomized method with two treatment periods of 8 weeks (placebo or candesartan 8 mg/day) and a wash-out period of 4 weeks in between. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were measured by inulin- and PAH-clearances at the beginning and the end of each treatment period. RESULTS: Proteinuria significantly decreased from 2 +/- 0.4 g/day to 1.3 +/- 0.3 g/day (P < 0.05) with the addition of candesartan treatment, whereas it remained unchanged (from 1.8 +/ 0.3 g/day to 1.9 +/- 0.3 g/day) under placebo. GFR (candesartan: from 66 +/- 13 to 58 +/- 11 ml/min per 1.73 m2, placebo: from 64 +/- 11 to 62 +/- 13 ml/min per 1.73 m2) and ERPF (candesartan: from 329 +/- 44 to 304 +/- 37 ml/min per 1.73 m2, placebo: from 362 +/- 48 to 315 +/- 46 ml/min per 1.73 m2) did not alter significantly after 8 weeks of treatment. The addition of candesartan treatment significantly reduced systolic blood pressure (from 129 +/- 3 to 123 +/- 2 mmHg, P < 0.05) and diastolic blood pressure (from 79 +/- 2 to 76 +/- 2 mmHg, P < 0.05) compared with placebo (systolic: 128 +/- 3 to 127 +/- 3 mmHg, diastolic: 79 +/- 2 to 79 +/- 2 mmHg). CONCLUSION: Candesartan promotes a complementary antiproteinuric and a small antihypertensive effect after a treatment period of 8 weeks in patients with chronic glomerulonephritis when given in conjunction with an ACEI. Renal hemodynamics did not vary significantly. PMID- 11910313 TI - Prevention of diastolic heart failure by endothelin type A receptor antagonist through inhibition of ventricular structural remodeling in hypertensive heart. AB - OBJECTIVES: Despite the clinical frequency of diastolic heart failure, its therapeutic strategy has not been established. Our recent study demonstrated activation of the endothelin (ET) system in a diastolic heart failure model with hypertension. Several studies have reported that ET type A (ETA) receptor antagonist improves systolic function and prevents systolic heart failure; however, its effects on diastolic heart failure are unknown. We investigated the effects of chronic administration of ET(A) receptor antagonist in diastolic heart failure. DESIGN AND METHODS: Dahl-Iwai salt-sensitive rats fed on a high-salt diet from 7 weeks of age, in which congestive heart failure develops following hypertension without cardiac chamber dilatation or systolic dysfunction, were divided into groups with and without administration of a subdepressor dose of ET(A) receptor antagonist. RESULTS: Hypertension induced compensatory left ventricular (LV) hypertrophy at 13 weeks in six untreated rats. Persistent pressure overload developed progressive LV hypertrophy and fibrosis from 13 to 19 weeks, resulting in elevated LV filling pressure and increased lung weight. Chronic ET(A) receptor blockade did not restrain compensatory LV hypertrophy at 13 weeks; however, it attenuated LV hypertrophy and fibrosis thereafter (n = 6). These beneficial effects resulted in the maintenance of normal LV filling pressure without changes in LV end-diastolic diameter, indicating prevention of LV stiffening. CONCLUSIONS: Chronic ET(A) receptor blockade is likely to exert beneficial effects in diastolic failure through attenuation of the progression of LV hypertrophy and fibrosis. PMID- 11910312 TI - Inhibition of left ventricular fibrosis by tranilast in rats with renovascular hypertension. AB - BACKGROUND: Growth factors such as transforming growth factor-beta (TGF beta) are believed to have an essential role in cardiac fibrosis. Tranilast (N(3,4 dimethoxycinnamoyl) anthranilic acid) attenuates the increased expression of TGF beta mRNA in vitro. OBJECTIVE: To investigate whether tranilast reduces cardiac fibrosis in rats with two-kidney, one-clip (2K1C) renovascular hypertension. In addition, we tested the in-vitro effects of tranilast on cardiac myocytes and non myocyte cells. METHODS: We analysed hearts from four groups of rats: sham operated controls; rats with 2K1C renovascular hypertension; rats with 2K1C renovascular hypertension treated for 12 weeks with the angiotensin converting enzyme (ACE) inhibitor, quinapril (6 mg/kg per day); rats with 2K1C renovascular hypertension treated for 12 weeks with tranilast (400 mg/kg per day). RESULTS: Systolic blood pressure was reduced after quinapril treatment. Tranilast did not alter blood pressure (2K1C: 223 +/- 19 mmHg; 2K1C + quinapril: 149 +/- 15 mmHg (P < 0.01 compared with 2K1C); 2K1C + tranilast: 204 +/- 32 mmHg). Left ventricular weight was likewise reduced significantly by quinapril, but not significantly by tranilast (2K1C: 1.52 +/- 0.2 g; 2K1C + quinapril: 1.26 +/- 0.18 g (P < 0.05 compared with 2K1C); 2K1C + tranilast: 1.37 +/- 0.27 g). Using a computer-aided image analysis system, we demonstrated that tranilast prevented cardiac fibrosis in a blood-pressure-independent manner (P < 0.01 compared with 2K1C). Determination of the cardiac hydroxyproline content similarly revealed a significant reduction in cardiac fibrosis by tranilast (2K1C: 4.92 +/- 0.48 mg/mg; 2K1C + tranilast: 3.97 +/- 0.46 mg/mg; P < 0.05). The effect of tranilast on cardiac fibrosis was comparable to the effects of a blood-pressure-decreasing dose of the ACE inhibitor, quinapril. Cell culture experiments revealed that tranilast significantly decreased the proliferation of cardiac non-myocyte cells. Proliferation of cardiac myocytes was not altered. CONCLUSION: This study revealed that long-term treatment with tranilast markedly attenuated left ventricular fibrosis in rats with renovascular hypertension. This was most probably the result of an antiproliferative effect of tranilast on cardiac non myocyte cells. Tranilast thus offers a unique new therapeutic approach to the reduction of TGF beta-mediated cardiac fibrosis in vivo. PMID- 11910314 TI - Attenuation of aortic banding-induced cardiac hypertrophy by propranolol is independent of beta-adrenoceptor blockade. AB - OBJECTIVE: Racemic propranolol attenuates cardiac hypertrophy secondary to abdominal aortic banding-induced pressure overload by a mechanism independent of its effect on cardiac work load. This was only observed, however, using doses of propranolol that were much higher than those needed to induce beta-adrenoceptor blockade. Thus, the question remains as to whether the antihypertrophic effect of propranolol depends on its ability to antagonize cardiac beta-adrenoceptor mediated action (positive chronotropic effect, trophic effect) or on beta adrenoceptor-independent action. METHODS: In a rat model of chronic pressure overload induced by abdominal aortic banding, we evaluated the effects on left ventricular hypertrophy (LVH) of the propranolol isomers, L-propranolol and D propranolol, which compared to L-isomer is approximately 50-fold less potent as a beta-adrenoceptor antagonist, but is similarly potent as a membrane-stabilizer, as well as of timolol, a non-selective beta-adrenergic antagonist devoid of membrane stabilizing activity, and disopyramide, which is a membrane stabilizer, but not a beta-adrenoceptor blocker. RESULTS: Compared to sham-operated rats, banded rats had 30% greater left ventricular to body weight (LVW/BW) ratio (P < 0.01). The increase in LVW/BW ratio was significantly attenuated by treatment with 40 and 80 (but not 10) mg/kg per day of L-propranolol. Left ventricular hypertrophy was also prevented by D-propranolol, 40 and 80 mg/kg per day, and disopyramide, 50 mg/kg per day, whereas timolol, 30 and 60 mg/kg per day, showed no antihypertrophic effect. In separate groups of banded rats in which the reduction in heart rate induced by propranolol (80 mg/kg per day) was prevented by chronic cardiac pacing at 375 b.p.m., hypertrophy was again prevented, indicating that the effects of L-propranolol on LVH are not related to a reduction in cardiac work load. CONCLUSIONS: In the aortic banding-induced model of LVH: (i) the antihypertrophic effect of propranolol is independent of its beta adrenergic blocking activity; and Iii) since disopyramide and D-propranolol also proved to be able to antagonize banding-induced LVH, the hypothesis is proposed that membrane-stabilizing activity, among the ancillary properties of propranolol, most likely accounts for the antihypertrophic effect of this drug. PMID- 11910315 TI - Double-blind, placebo-controlled crossover comparison of five classes of antihypertensive drugs. AB - OBJECTIVE: Hypertension guidelines recommend initial treatment with a beta blocker or diuretic and adding the other drug where blood pressure is not controlled. We hypothesized that systematic rotation through the major classes of antihypertensive drugs would demonstrate substantial differences in the pattern of an individual patient's response, and suggest a more rational approach to choosing best treatment. DESIGN: Thirty-four young hypertensives (age 28-55, median 47) rotated in a double-blind, Latin-square, crossover fashion through 6 weeks of treatment each with amlodipine, doxazosin, lisinopril, bisoprolol, bendrofluazide and placebo. Blood pressure was measured at each visit. 'Best' drug, defined by efficacy and tolerability, was repeated at the end. RESULTS: Rotation doubled the number of patients reaching target blood pressure (systolic < 140 mmHg) on one drug (P = 0.03). All five drugs were represented among the 'best' drugs. In six patients, the blood pressure on 'best' drug was at least 10 mmHg lower than on any other. Response to the 'best' drug was highly correlated (r = 0.79) with its previous administration. By contrast, there were only weak correlations between responses to pairs of drugs, except for angiotensin converting enzyme (ACE) inhibitor (A) with beta-blocker (B), and calcium blocker (C) with diuretic (D) - each r = 0.71, P < 0.005). In these young patients, the majority of patients (23/34) responded best to a drug suppressing the renin system (A and B). CONCLUSIONS: Patients vary reproducibly in their response to initial treatment, and switching among drugs can increase the efficacy of monotherapy. The results support an AB/CD scheme for choosing therapy, in which the first drug is taken from one of these pairs, and uncontrolled patients switch to one of the other pair. PMID- 11910318 TI - Dr. Charlie: the insights (and occasional errors) of a quondam oculoplastic surgeon. PMID- 11910316 TI - Hypertension in high-risk patients: beware of the underuse of effective combination therapy (results of the PRATIK study). AB - OBJECTIVE: To determine whether blood pressure control in a general practice setting is influenced by the presence of additional risk factors, and to analyse the role of antihypertensive therapy in this relationship. DESIGN: A cross sectional study was conducted with a sample of 3153 general practitioners. SETTING: Primary care. PARTICIPANTS: The first five hypertensive patients presenting at the practitioner's office were included. MAIN OUTCOME MEASURES: Cardiovascular risk factors, antihypertensive drugs and cardiovascular history were reported. Blood pressure was measured. The analysis was conducted in treated patients who were divided in three groups: no other risk factors (group I); 1-2 risk factors (group II); 3 or more risk factors or target-organ damage or diabetes or associated cardiovascular disease (group III). RESULTS: Data were available for all variables in the 14 066 treated hypertensive individuals who form the basis of this report. Blood pressure control had been achieved in a lower percentage of individuals in group III (27%) than in group I (42.9%). To control hypertension, combination therapies were more frequently required in group III (55.8%) than in group II (43.5%) or group I (34.2%). Among individuals with uncontrolled hypertension, about 39% of patients in group III were receiving monotherapy and the percentage receiving two-drug treatments identified as effective in the 1999 WHO guidelines was significantly lower in group III. CONCLUSION: The study shows that, in general practice, control of blood pressure decreases as the number of risk factors present increases. An underuse of combination therapy, especially effective two-drug treatment in patients with several risk factors, may account for this finding. PMID- 11910319 TI - Stretching of the Mueller muscle results in involuntary contraction of the levator muscle. AB - PURPOSE: Since the levator muscle involuntarily and tonically contracts against the weight and elastic resistance of the upper eyelid to maintain an adequate visual field, a mechanoreceptor such as a muscle spindle or a periodontal mechanoreceptor is thought to be essential for its functioning. It was surmised that the Mueller muscle might act as a serial kind of muscle spindle of the levator muscle. METHODS: The response of the bilateral levator muscles evoked by stretching the Mueller muscle of each eyelid of 87 patients with dermatochalasis or aponeurotic blepharoptosis was electromyographically and photographically recorded. RESULTS: Stretching of the unilateral Mueller muscle evoked contraction of the ipsilateral levator muscle in 18 and of the bilateral levator muscle in 69 of the 87 patients. CONCLUSIONS: The Mueller muscle can be thought of as a large, serial kind of muscle spindle, so that stretching by voluntary phasic contraction of the levator muscle for initial eye opening may evoke an afferent impulse to the mesencephalic trigeminal nucleus. Subsequently, this nucleus may stimulate the central caudal nucleus of the oculomotor nuclear complex, leading to involuntary tonic contraction of the ipsilateral or bilateral levator muscles, in the form of a continuous stretch reflex, to maintain an adequate visual field. PMID- 11910322 TI - Combined cervicofacial rhytidectomy and laser skin resurfacing. AB - PURPOSE: To determine the safety and efficacy of simultaneous cervicofacial rhytidectomy (face lift) and laser skin resurfacing. METHODS: A retrospective study of 100 consecutive patients who underwent simultaneous face lift and carbon dioxide laser resurfacing procedures was performed. Patients received regional (periorbital and/or perioral), "T"-shaped (forehead and central face), or full face laser skin resurfacing, and were classified on this basis. All patients were assessed for flap necrosis, delayed reepithelialization, reactive hyperpigmentation, persistent erythema, scarring, or other unsatisfactory results postoperatively. RESULTS: One of 10 patients receiving full-face resurfacing, including the entire subcutaneously undermined flap, suffered full-thickness skin necrosis of the distal segment of one flap. Six of the 100 patients developed reactive hyperpigmentation postoperatively. One patient evidenced an imprint of the laser footprint over nonundermined skin, requiring a secondary procedure. Aside from the one patient with scar, no cases of delayed reepithelialization were identified. CONCLUSIONS: Carbon dioxide laser skin resurfacing combined with face lift procedures can achieve dramatic cervicofacial rejuvenation. Laser resurfacing over nonundermined skin during the face lift procedure does not appear to increase the risk of postoperative complication. Laser treatment over an undermined distal face lift flap appears to increase the potential for skin necrosis, and should not be considered a routine modality for facial rejuvenation. PMID- 11910321 TI - Eyelid lymphatics I: histochemical comparisons between the monkey and human. AB - PURPOSE: To study the lymphatic drainage of the cynomolgus monkey and human eyelid and periocular tissue by means of histochemistry. METHODS: Eyelid and periocular tissue from three cynomolgus monkeys undergoing sacrifice for glaucoma and retina research purposes and discarded tissue from a wedge resection of one human eyelid were used for histochemical analysis. Lymphatic capillaries were distinguished histochemically in monkey and human eyelids by light microscopy with a 5'-nucleotidase (5'-Nase) staining method. Identification of lymphatic vessels was based on strict morphologic criteria combined with specific 5'-Nase staining. RESULTS: Histochemical analysis with 5'-nucleotidase revealed a subcutaneous and pretarsal lymphatic plexus in both the human and monkey. CONCLUSIONS: Histochemical results demonstrate similar lymphatic plexi in the monkey and human. Future studies will help to clarify the lymphatic drainage pathways of monkey and human eyelids. PMID- 11910320 TI - Clinicopathologic evaluation of the Mueller muscle in thyroid-associated orbitopathy. AB - PURPOSE: To study the histopathologic features of the Mueller muscle in chronic eyelid retraction caused by thyroid-associated orbitopathy. To investigate if the degree of eyelid retraction correlates with any histopathologic finding. METHODS: A prospective case series of 23 consecutive patients with thyroid-associated orbitopathy was studied. Specimens were obtained during a standard muellerectomy. Formalin-preserved specimens were studied with the use of hematoxylin-eosin, periodic acid-Schiff, Masson trichrome, and Giemsa stains. Immunostaining against leukocyte common antigen, L26, CD3, and KP-1 was performed. Three control specimens were also evaluated in a similar fashion. Fresh tissue was placed in cold glutaraldehyde overnight, postfixed, dehydrated, and infiltrated with epoxy resin. Silver (70 nm) sections were cut and stained with uranyl acetate and lead citrate for electron microscopic examination. RESULTS: On light microscopy, fibrosis and mast cell infiltration was present in all 23 specimens. Fat infiltration was noted in 16 of 23 specimens and did not correlate with increasing age of the patient. Interstitial edema and lymphocytic infiltration were not observed. On immunohistochemistry, leukocyte common antigen was positive, confirming the presence of inflammation. L26, CD3, and KP1 were negative. Electron microscopy demonstrated fibrosis, mast cells, and abundant contracting Mueller cells. The degree of clinical retraction in millimeters did not correlate with fibrosis, inflammation, or fat infiltration. The control specimens demonstrated rare fat and mast cell infiltration and no fibrosis. CONCLUSIONS: Contrary to previous reports, the Mueller muscle is involved in the inflammation and fibrosis that characterizes thyroid-associated orbitopathy. The Mueller muscle is grossly enlarged. On histopathologic inspection, fibrosis, fatty infiltration, and increased mast cell presence accompany focal atrophy of the Mueller muscle. In concordance with prior descriptions, many Mueller cells are in an actively contracting state on electron microscopy. PMID- 11910323 TI - No-scar Asian epicanthoplasty: a subcutaneous approach. AB - PURPOSE: The epicanthal fold is a normal finding in the medial portion of the upper eyelid in many Asians. We describe a simple technique of removing the excess muscle and softening or eliminating the epicanthal fold without making incisions in the medial canthal region, thus avoiding complications such as scarring or web formation METHODS: Interventional case series. The subcutaneous epicanthoplasty was performed on all Asian patients undergoing concurrent upper eyelid blepharoplasty or aponeurotic ptosis repair. RESULTS: Thirty-eight Asian patients underwent epicanthal fold correction between January 1996 and December 2000. All patients had softening of the epicanthal fold; however, some cases of mild undercorrection were noted. CONCLUSIONS: Softening or elimination of the Asian epicanthal fold can be accomplished without making skin incisions in the medial canthal region. Our technique is a simple, graded procedure that can be performed in conjunction with upper blepharoplasty or ptosis repair. PMID- 11910324 TI - Eyelid sensation after supratarsal lid crease incision. AB - PURPOSE: To determine the severity and duration of the loss of eyelid sensation after upper eyelid crease incision. METHODS: This clinic-based case study was performed by analyzing observational measurements of patients undergoing upper blepharoplasty or ptosis surgery. Eighty-three eyelids of 50 patients were studied. A Cochet-Bonnet filament-type aesthesiometer was used to obtain all measurements. Preoperative and postoperative measurements were recorded at 1 week, 1 month, and final (2-6 months) time periods. Statistical analysis evaluated the degree and duration of the sensory loss and the extent of recovery during the evaluation period. Recovery of sensation was defined as a numerical reading within one point of baseline. RESULTS: The mean aesthesiometry reading was calculated at the preoperative (3.45), 1-week (1.20), 1-month (1.56), and final postoperative (2.56) periods. Paired t testing showed a decreased but significant difference in sensation measurement at each comparison. Recovery of sensation to within one point occurs at the preoperative to late time period comparison. All but 4 of the 68 eyelids tested at the 1-week postoperative time period had a measured loss of sensation. Of the 44 eyelids tested at the final time period, all but 1 had regained some or all of this sensory loss. CONCLUSIONS: Loss of skin sensation in the eyelid after upper eyelid crease incision blepharoplasty or blepharoptosis repair occurs in most patients and should be considered an expected outcome of the procedure. Partial to complete recovery of eyelid sensation over 2 to 6 months should also be expected, though in rare instances this does not occur. PMID- 11910325 TI - The quasi-integrated porous polyethylene orbital implant. AB - PURPOSE: To describe a new quasi-integrated porous polyethylene orbital implant that combines the advantages of host tissue incorporation and improved motility with a single-stage surgery. METHODS: Twenty-four consecutive patients undergoing primary or secondary orbital implantation received the quasi-integrated porous polyethylene implant. Approximately 6 weeks after implantation, a custom-fitted prosthesis was made by an impression technique to provide a "lock-and-key" fit with the orbital implant. Postoperative complications and motility of the prosthetic shell were evaluated. RESULTS: During the 27-month period between December 1998 and March 2001, 24 patients received the quasi-integrated porous polyethylene implant as a buried orbital implant. Thirteen patients received the implant as a primary orbital implant after either evisceration or enucleation and 11 patients received the implant as a secondary orbital implant. Follow-up ranged from 3 months to 30 months, with an average of 16.9 months. All patients were considered to have good motility of their prosthetic shell at their final follow up visit. No cases of implant extrusion or migration were noted. Two patients required deepening of their inferior fornix to accommodate the increased motility of their prosthesis. CONCLUSIONS: The new quasi-integrated porous polyethylene orbital implant provides improved motility without the need for secondary placement of pegs or screws. It has the advantage of biocompatibility, allowing host tissue incorporation to resist implant migration and extrusion. The implant is available in three sizes: small, medium, and large, approximating the volume of a 16-, 18-, and 20-millimeter sphere, respectively. PMID- 11910326 TI - Acquired monocular vision: functional consequences from the patient's perspective. AB - PURPOSE: The study is conducted to determine the effect of acquired monocular vision (enucleation, phthisis) on the daily activities of patients. METHODS: Sixty-five patients in a case series completed a 30-question survey evaluating the functional consequences of monocular vision. RESULTS: Patients with sudden visual loss adapted more slowly than those with gradual loss (8.8 versus 3.6 months). Difficulties with depth perception and cosmesis were prevalent. Unexpected consequences included neck pain, strain with reading, employment change, depression, car accidents, and alcoholism. Ninety-one percent of patients had no formal training to help them adapt. CONCLUSIONS: More than previously appreciated, there are many obstacles that patients face in making the transition to monocular vision. Few patients receive any formal instruction to help them adapt, yet many indicate that it would be beneficial. PMID- 11910327 TI - Orbital invasion with prolactinoma: a clinical review of four patients. AB - PURPOSE: To present four patients with invasive prolactin-secreting pituitary adenomas extending in the orbit. METHODS: Retrospective, noncomparative case series. RESULTS: In all cases, the tumor was a large, invasive prolactin secreting adenoma. In three cases, significant permanent vision loss occurred and severe disfigurement resulted; two of these patients died of tumor-related complications. In one case, the tumor responded well to medical treatment, and the orbital symptoms improved significantly. CONCLUSION: Although it is rare for a pituitary adenoma to invade the orbit, devastating consequences to the integrity of the globe and ocular adnexa may result. Therefore, early recognition and proper treatment of orbital pituitary tumors are of utmost importance to minimize ocular and orbital damage. PMID- 11910328 TI - Advanced magnetic resonance imaging techniques for patients with hemifacial spasm. AB - PURPOSE: To review the underlying causes, diagnostic issues, and treatment of hemifacial spasm, with emphasis on advanced MRI techniques. METHODS: Brief technical note. RESULTS: High-resolution T1- and T2-weighted spin-echo and/or gradient echo imaging of the posterior fossa should be performed with the use of intravenous gadolinium for maximum contrast between CSF, vessel, and nerve. Magnetic resonance angiography is often useful, and new state-of-the-art sequences provide more detail. CONCLUSIONS: As MRI techniques improve, diagnosis and treatment of patients with hemifacial spasm will become easier. Ophthalmologists should be aware of these new magnetic resonance techniques. PMID- 11910329 TI - Congenital retraction of the lower eyelid: two case reports. AB - PURPOSE: Idiopathic congenital retraction of the lower eyelid is rare. To further define the clinical features of this condition, we describe two new cases. METHODS: Retrospective chart review. RESULTS: In both cases, eye movements were restricted ipsilaterally and the condition was nonprogressive. CT scans showed mildly enlarged inferior and medial rectus muscles in one patient and normal muscles in the other. All other orbital structures were normal. CONCLUSIONS: No definite cause for the lower eyelid retraction could be found, but it appeared to be related to a tight inferior rectus muscle. These patients may represent a variant of ocular fibrosis syndrome. PMID- 11910330 TI - Lower eyelid reverse ptosis repair. AB - PURPOSE: To report the results of the surgical repair of lower eyelid reverse ptosis. METHODS: Retrospective case series. Eight patients ranging in age from 31 to 77 years underwent surgical repair of lower eyelid reverse ptosis. The pupillary axis of the affected eye(s) in each patient was obscured in downgaze, interfering with reading. The lower eyelid reverse ptosis resulted from involutional changes in 3 patients, previous orbital decompression in 3 patients, multiple prior retinal and extraocular muscle operations in 1 patient, and previous orbital floor fracture and repair in 1 patient. Transcutaneous advancement of the lower eyelid retractors was performed in 12 eyelids of the 8 patients. RESULTS: The mean preoperative vertical eyelid fissure was 6.2 mm (median, 6 mm; range, 3-9 mm), increasing after surgery to a mean of 7.7 mm (median, 8 mm; range, 5-11 mm). The mean preoperative distance between the central light reflex and the lower eyelid margin was 1.7 mm (median, 1.25 mm; range, 1-4 mm); this distance increased to a mean of 3.3 mm (median, 3.25 mm; range, 2.5-4.5 mm) after surgery. Symptoms improved in all patients, and there were no perioperative complications. Follow-up intervals ranged from 2 months to 24 months (mean, 9 months; median, 13 months). CONCLUSIONS: Analogous to upper eyelid ptosis repair by advancement of the levator aponeurosis, lower eyelid reverse ptosis may be corrected effectively and safely by advancing the lower eyelid retractors. PMID- 11910331 TI - Nonpalpable breast carcinoma presenting as orbital infiltration: case presentation and literature review. AB - PURPOSE: To report orbital metastasis as the presenting sign of a nonpalpable breast carcinoma in a 61-year-old woman. METHODS: Case report and literature review. RESULTS: A uterine metastasis of unknown origin found 11 years prior was retrospectively identified as having the same histopathology as the breast and orbit tumors. Eight years after her orbital presentation, the patient was diagnosed with and underwent resection of a metastatic adenocarcinoma of the colon, consistent with lobular adenocarcinoma of the breast. CONCLUSIONS: This is a rare case of an orbital metastatic adenocarcinoma preceding the diagnosis of a nonpalpable primary breast carcinoma. PMID- 11910332 TI - Ovarian cancer as a chronic disease: a new treatment paradigm. Highlights of a roundtable discussion. PMID- 11910333 TI - Nursing perspectives on patient management during chronic therapy for relapsed ovarian cancer. PMID- 11910334 TI - The best and the brightest. PMID- 11910336 TI - The molecular classification of the clinical manifestations of Crohn's disease. AB - BACKGROUND & AIMS: Crohn's disease is a common inflammatory disorder of the gut characterized by variation in both location and behavior. Chromosome 16 and the HLA region on chromosome 6 have been implicated in susceptibility to disease. Mutations in the NOD2/CARD15 gene, recently identified on chromosome 16, have been associated with disease overall but are found in only 25% of patients. No data regarding their contribution to specific disease subtypes exist. Here we report a detailed genotype-phenotype analysis of 244 accurately characterized patients. METHODS: A total of 244 white patients with Crohn's disease recruited from a single center in the United Kingdom were studied. All patients were rigorously phenotyped and followed-up for a median time of 16 years. By using linkage disequilibrium mapping we studied 340 polymorphisms in 24 HLA genes and 3 NOD2/CARD15 polymorphisms. RESULTS: We show that NOD2/CARD15 mutations determine ileal disease only. We confirm that alleles on specific long-range HLA haplotypes determine overall susceptibility and describe novel genetic associations with susceptibility, location, and behavior of Crohn's disease. CONCLUSIONS: The clinical pattern of Crohn's disease may be defined by specific genotypes. This study may provide the basis for a future molecular classification of disease. PMID- 11910337 TI - The contribution of NOD2 gene mutations to the risk and site of disease in inflammatory bowel disease. AB - BACKGROUND & AIMS: Mutations in the NOD2 gene are strongly associated with susceptibility to Crohn's disease (CD). We analyzed a large cohort of European patients with inflammatory bowel disease to determine which mutations confer susceptibility, the degree of risk conferred, their prevalence in familial and sporadic forms of the disease, and whether they are associated with site of disease. METHODS: Individuals were genotyped for 4 NOD2 mutations: P268S, R702W, G908R, and 3020insC. Allelic transmission distortion to 531 CD- and 337 ulcerative colitis-affected offspring was assessed by the transmission disequilibrium test. Association was also tested in an independent cohort of 995 patients with inflammatory bowel disease and 290 controls. Cases were stratified by disease site and compared across NOD2 genotypes. RESULTS: R702W, G908R, and 3020insC were strongly associated with CD but not with ulcerative colitis. Linkage disequilibrium was observed between P268S and the other mutations, forming 3 independent disease haplotypes. Genotype relative risks were 3.0 for mutation heterozygotes and 23.4 for homozygotes or compound heterozygotes. The frequency of NOD2 mutations was higher in cases from families affected only with CD and was significantly increased in ileal-specific disease cases compared with colon-specific disease (26.9% vs. 12.7%, P = 0.0004). CONCLUSIONS: The R702W, G908R, and 3020insC mutations are strong independent risk factors for CD and are associated particularly with ileal disease. PMID- 11910338 TI - The natural history of fistulizing Crohn's disease in Olmsted County, Minnesota. AB - BACKGROUND & AIMS: Little is known about the cumulative incidence and natural history of fistulas in Crohn's disease in the community. METHODS: The medical records of all Olmsted County, Minnesota residents who were diagnosed with Crohn's disease from 1970 to 1993 and who developed a fistula were abstracted for clinical features and outcomes. Six patients denied research authorization. The cumulative incidence of fistula from time of diagnosis was estimated by using the Kaplan-Meier product-limit method. RESULTS: At least 1 fistula occurred in 59 patients (35%), including 33 patients (20%) who developed perianal fistulas. Twenty-six (46%) developed a fistula before or at the time of formal diagnosis. Assuming that the 9 patients with fistula before Crohn's disease diagnosis were instead simultaneous diagnoses, the cumulative risk of any fistula was 33% after 10 years and was 50% after 20 years (perianal, 21% after 10 years and 26% after 20 years). At least 1 recurrent fistula occurred in 20 patients (34%). Most fistulizing episodes (83%) required operations, most of which were minor. However, 11 perianal fistulizing episodes (23%) resulted in bowel resection. CONCLUSIONS: Fistulas in Crohn's disease were common in the community. In contrast to referral-based studies, only 34% of patients developed recurrent fistulas. Surgical treatment was frequently required. PMID- 11910339 TI - Celiac disease in patients with severe liver disease: gluten-free diet may reverse hepatic failure. AB - BACKGROUND & AIMS: Mild liver abnormalities are common in patients with celiac disease and usually resolve with a gluten-free diet. We investigated the occurrence of celiac disease in patients with severe liver failure. METHODS: Four patients with untreated celiac disease and severe liver disease are described. Further, the occurrence of celiac disease was studied in 185 adults with previous liver transplantation using serum immunoglobulin A endomysial and tissue transglutaminase antibodies in screening. RESULTS: Of the 4 patients with severe liver disease and celiac disease, 1 had congenital liver fibrosis, 1 had massive hepatic steatosis, and 2 had progressive hepatitis without apparent origin. Three were even remitted for consideration of liver transplantation. Hepatic dysfunction reversed in all cases when a gluten-free diet was adopted. In the transplantation group, 8 patients (4.3%) had celiac disease. Six cases were detected before the operation: 3 had primary biliary cirrhosis, 1 had autoimmune hepatitis, 1 had primary sclerosing cholangitis, and 1 had congenital liver fibrosis. Only 1 patient had maintained a long-term strict gluten-free diet. Screening found 2 cases of celiac disease, 1 with autoimmune hepatitis and 1 with secondary sclerosing cholangitis. CONCLUSIONS: The possible presence of celiac disease should be investigated in patients with severe liver disease. Dietary treatment may prevent progression to hepatic failure, even in cases in which liver transplantation is considered. PMID- 11910340 TI - The association between hepatitis C infection and survival after orthotopic liver transplantation. AB - BACKGROUND & AIMS: The effect of hepatitis C viral (HCV) infection on patient and allograft survival after orthotopic liver transplantation is controversial. Hepatitis C recurrence after transplant is inevitable, but studies to date have not found a survival difference between recipients with and without HCV. METHODS: Using data from the United Network for Organ Sharing, we performed a retrospective cohort study of 11,036 patients who underwent 11,791 liver transplants between 1992 and 1998. The hazard rates of patient and allograft survival for patients who were HCV-positive as compared with patients who were HCV-negative were assessed by proportional-hazards analysis, with adjustment for potential confounding variables, including donor, recipient, and transplant center characteristics. RESULTS: Liver transplantation in HCV-positive recipients was associated with an increased rate of death (hazard ratio, 1.23; 95% confidence interval [CI], 1.12-1.35) and allograft failure (hazard ratio, 1.30; 95% CI, 1.21-1.39), as compared with transplantation in HCV-negative recipients. This reduction in survival persisted after adjusting for potential confounders. There was an interaction between HCV and sex (P < 0.001) with the effect of HCV on survival being most pronounced in female recipients (patient survival hazard ratio, 1.56; 95% CI, 1.35-1.81; allograft survival hazard ratio, 1.51; 95% CI, 1.34-1.70). CONCLUSIONS: HCV infection significantly impairs patient and allograft survival after liver transplantation. PMID- 11910341 TI - Occurrence of hepatopulmonary syndrome in Budd-Chiari syndrome and the role of venous decompression. AB - BACKGROUND & AIMS: Hepatopulmonary syndrome (HPS) has been predominantly detected in cirrhotic patients and rarely in patients with noncirrhotic portal hypertension. The aim of this study was to determine the occurrence of HPS in patients with Budd-Chiari syndrome (only anecdotal reports available) and evaluate the role of venous decompression in its reversal. METHODS: Twenty-nine consecutive cases of Budd-Chiari syndrome without primary cardiopulmonary disease were investigated by air contrast echocardiography and arterial blood gas analysis. Venous decompression (e.g., by balloon cavoplasty) was attempted when feasible. RESULTS: Eight cases (27.6%) of HPS and 9 cases (31.0%) with positive contrast echocardiography but unimpaired oxygenation were detected. Duration of disease was longer (P = 0.026) among those with positive contrast echocardiography. Cavoplasty reversed 4 of 5 cases of HPS and 2 of 2 cases with positive contrast echocardiography alone. Venous decompression by drainage of amebic liver abscess (which was compressing hepatic venous outflow) also reversed 1 case of HPS. HPS was relieved by venous decompression in 5 of 6 cases. CONCLUSIONS: HPS developed in a substantial fraction of our patients with Budd Chiari syndrome, with positive contrast echocardiography occurring mainly in the benign, slowly progressing variety. Venous decompression showed promise in reversing such cases. PMID- 11910342 TI - 6-MP metabolite profiles provide a biochemical explanation for 6-MP resistance in patients with inflammatory bowel disease. AB - BACKGROUND & AIMS: Approximately 40% of inflammatory bowel disease (IBD) patients fail to benefit from 6-mercaptopurine (6-MP)/azathioprine (AZA). Recent reports suggest 6-thioguanine nucleotide (6-TGN) levels (>235) independently correlate with remission. An inverse correlation between 6-TGN and thiopurine methyltransferase (TPMT) has been described. The objectives of this study were to determine whether dose escalation optimizes both 6-TGN levels and efficacy in patients failing therapy because of subtherapeutic 6-TGN levels and its effect on TPMT. METHODS: Therapeutic efficacy and adverse events were recorded at baseline and upon reevaluation after dose escalation in 51 IBD patients. 6-MP metabolite levels and TPMT activity were recorded blinded to clinical information. RESULTS: Fourteen of 51 failing 6-MP/AZA at baseline achieved remission upon dose escalation, which coincided with significant rises in 6-TGN levels. Despite increased 6-MP/AZA doses, 37 continued to fail therapy at follow-up. Dose escalation resulted in minor changes in 6-TGN, yet a significant increase in 6 methylmercaptopurine ribonucleotides (6-MMPR) (P < or = 0.01) and 6-MMPR:6-TGN ratio (P < 0.001). 6-MMPR rises were associated with dose-dependent hepatotoxicity in 12 patients (24%). TPMT was not influenced by dose escalation. CONCLUSIONS: Serial metabolite monitoring identifies a novel phenotype of IBD patients resistant to 6-MP/AZA therapy biochemically characterized by suboptimal 6-TGN and preferential 6-MMPR production upon dose escalation. PMID- 11910343 TI - Effects of long-term propranolol and octreotide on postprandial hemodynamics in cirrhosis: a randomized, controlled trial. AB - BACKGROUND & AIMS: Postprandial increases in portal pressure may influence esophageal variceal rupture. The effects of chronic propranolol and octreotide (100 and 200 microg subcutaneously in a single dose) on postprandial hemodynamics were evaluated. METHODS: FIRST STUDY: 36 cirrhotic patients were studied at baseline and 30 and 60 minutes after a standard meal and then treated with propranolol (139 +/- 9 mg/d during 39 +/- 2 days). SECOND STUDY: After baseline measurements, patients were randomized into 3 groups: (1) placebo, (2) octreotide (100 microg), or (3) octreotide (200 microg) (n = 12 for each group). Thirty minutes postinjection a new baseline was established and measurements were repeated 30 and 60 minutes after the meal. RESULTS: First study: Baseline portal pressure was 18.1 +/- 1.2 mm Hg, 30 and 60 minutes after the meal it was 21.5 +/- 0.8 mm Hg and 20.5 +/- 0.8 mm Hg, respectively (both P < 0.01 vs. baseline). Cardiac index (CI) was 4.5 +/- 0.2, 4.8 +/- 0.2, and 4.9 +/- 0.2 L x min(-1) x m( 2), respectively (both P < 0.05 vs. baseline). Peripheral vascular resistance was 1012 +/- 56, 902 +/- 51 (P = NS), and 884 +/- 49 dynes x sec x cm(-5) (P< 0.05 vs. baseline), respectively. Second study: Propranolol and placebo did not blunt postprandial increase in portal pressure. Octreotide (100 microg) partially ameliorated postprandial increase in portal pressure. Octreotide (200 microg) significantly enhanced the portal hypotensive effect of propranolol and blunted the postprandial increase in portal pressure. CONCLUSIONS: Octreotide blunts postprandial increase in portal pressure not prevented by long-term propranolol administration. PMID- 11910344 TI - Terlipressin in patients with cirrhosis and type 1 hepatorenal syndrome: a retrospective multicenter study. AB - BACKGROUND & AIMS: Type 1 hepatorenal syndrome (HRS) is a severe complication of cirrhosis associated with a short median survival time (<2 weeks). Although the administration of terlipressin improves renal function, its effect on survival is unknown. This study investigated predictive factors of survival in patients with type 1 HRS treated with terlipressin. METHODS: Ninety-nine patients with type 1 HRS treated with terlipressin in 24 centers were retrospectively studied. Terlipressin-induced improved renal function was defined as a decrease in serum creatinine value to <130 micromol/L or a decrease of at least 20% at the end of treatment. RESULTS: At inclusion, the Child-Pugh score was 11.8 +/- 1.6 (mean +/- SD). Terlipressin (3.2 +/- 1.3 mg/day) was administered for 11 +/- 12 days. Renal function improved in 58% of patients (serum creatinine decreased by 46% +/- 17% from 272 +/- 114 micromol/L). Median survival time was 21 days. Survival rate was 40% at 1 month. Multivariate analysis showed that improved renal function and Child-Pugh score < or =11 at inclusion were independent predictive factors of survival (P < 0.0001 and 0.02, respectively). Thirteen patients underwent liver transplantation (92 +/- 95 days after HRS onset), 10 of whom had received terlipressin and had had improved renal function. CONCLUSIONS: This retrospective uncontrolled study shows that in patients with type 1 HRS, terlipressin-induced improved renal function is associated with an increase in survival. Thus, a randomized trial investigating the effect of terlipressin on survival in patients with type 1 HRS should be performed. PMID- 11910345 TI - Effect of iron depletion in carbohydrate-intolerant patients with clinical evidence of nonalcoholic fatty liver disease. AB - BACKGROUND & AIMS: Increased body iron, genetic hemochromatosis (GH) mutations, and nonalcoholic fatty liver disease (NAFLD) tend to cluster in carbohydrate intolerant patients. In an attempt to further clarify the interrelationships among these conditions, we studied 42 carbohydrate-intolerant patients who were free of the common GH mutations C282Y and H63D, and had a serum iron saturation lower than 50%. METHODS: We measured body iron stores, and induced iron depletion to a level of near-iron deficiency (NID) by quantitative phlebotomy. RESULTS: In the 17 patients with clinical evidence of NAFLD, we could not demonstrate supranormal levels of body iron (1.6 +/- 0.2 vs. 1.4 +/- 0.2 g; P = 0.06). However, at NID, there was a 40%-55% improvement (P = 0.05-0.0001) of both fasting and glucose-stimulated plasma insulin concentrations, and near normalization of serum alanine aminotransferase activity (from 61 +/- 5 to 32 +/- 2 IU/L; P < 0.001). CONCLUSIONS: These results reflect the insulin-sparing effect of iron depletion and indicate a key role of iron and hyperinsulinemia in the pathogenesis of NAFLD. PMID- 11910347 TI - Dietary sphingomyelin suppresses intestinal cholesterol absorption by decreasing thermodynamic activity of cholesterol monomers. AB - BACKGROUND & AIMS: In humans, cholesterol absorbed from the intestine contributes appreciably to serum cholesterol levels. We hypothesized that cholesterol thermodynamic activity (A(t)) would predict bioavailability of cholesterol monomers in intestinal content, and that natural dietary phospholipids exhibiting high affinity for cholesterol would reduce its absorption. METHODS: Cholesterol A(t) was determined by measuring partitioning of monomeric cholesterol from aqueous solutions of taurocholate, cholesterol, and either milk sphingomyelin (MSM), dipalmitoyl phosphatidylcholine (DPPC), or egg yolk phosphatidylcholine (EYPC) into wafers of polymerized silicone. Cholesterol absorption from the same mixtures was tested with monolayers of Caco-2 cells. For in vivo absorption studies (employing male C57L/J mice), we used the fecal dual isotope method during dietary enrichment with MSM, DPPC, or EYPC at varying dose levels. RESULTS: Cholesterol A(t) values were reduced significantly in MSM- and DPPC containing systems compared with EYPC and correlated positively with reduced uptake and esterification of cholesterol by Caco-2 cells. Mice fed chow absorbed 31.4% +/- 6.9% (mean +/- SEM) cholesterol, whereas enrichment with MSM or DPPC led to dose-dependent decreases in cholesterol absorption; even at 0.1% MSM, cholesterol absorption was reduced by 20.4% +/- 15.4% (P < 0.05, n = 6). CONCLUSIONS: Different phospholipids have distinct effects on micellar cholesterol A(t), which predicts cholesterol uptake by enterocytes in vitro as well as in vivo. Natural phospholipids with high affinity for cholesterol, as evidenced particularly by sphingomyelin, decrease A(t) and curtail intestinal cholesterol absorption. PMID- 11910346 TI - Hereditary nonpolyposis colorectal cancer in young colorectal cancer patients: high-risk clinic versus population-based registry. AB - BACKGROUND & AIMS: Early onset colorectal cancer (CRC) is an important feature of hereditary nonpolyposis colorectal cancer (HNPCC). We sought to compare rates of genetically defined HNPCC among individuals with early onset CRC drawn from a high-risk clinic and a population-based cancer registry. METHODS: Probands with CRC diagnosed before 36 years of age were enrolled from a high-risk CRC clinic at the University of California, San Francisco (UCSF), and a population-based Kaiser Permanente (KP) Health Plan cancer registry. Probands provided cancer family histories and tumors for microsatellite instability (MSI) testing and MSH2/MLH1 protein immunostaining. Germline MSH2 and MLH1 mutational analysis was performed. RESULTS: Forty-three probands were enrolled from UCSF and 23 from KP. The UCSF and KP probands had similar median age of onset of CRC (30 vs. 31 years) and the percentage with any personal or family history of another HNPCC-related cancer (70% vs. 74%). However, 28 of 40 (70%) of the UCSF tumors were MSI-H compared with 6 of 18 (33%) of KP tumors (P = 0.01), and 13 germline MSH2 or MLH1 mutations were found in the UCSF group compared with 0 in the KP group (P = 0.0001). In a multivariate analysis, institution (P = 0.002) and the total number of colorectal cancers in the family (P = 0.0001) were independent predictors of MSH2 or MLH1 mutation. CONCLUSIONS: Family history of cancer is an important feature of HNPCC, even among individuals with early onset CRC. Caution must be undertaken when extrapolating data regarding HNPCC from high-risk clinic populations to the general population. PMID- 11910348 TI - Ivermectin used in percutaneous drug injection method for the treatment of liver hydatid disease in sheep. AB - BACKGROUND & AIMS: Ivermectin is a macrocyclic lactone (avermectins) produced by the actinomycete Streptomyces avermitilis. In this experimental study, the effectiveness of intracystic injection of ivermectin was studied as a new approach of percutaneous treatment of cystic echinococcosis. METHODS: Twelve naturally infected sheep were selected and divided into 2 subgroups: treatment group (n = 9) and control group (n = 3). In the treatment group, approximate volume of ivermectin solution needed to achieve an intracystic concentration of 10 microg/mL was injected into cysts, but in the control group, sterile distillated water was applied. No reaspiration was performed at all. RESULTS: In the following period of 6 months, repeated sonography revealed a significant decrease in cyst sizes and progressive solidification of the cysts in the treatment group. In the control group, volumes of the cysts were increased. No major complications occurred during or after the procedure. After 6 months, all sheep were killed and examined for macroscopic and microscopic changes. Pathologic examination in the treatment group showed pericyst hyalinization, inflammatory cells in the cyst wall, degeneration of laminated and germinal membrane, and necrotic material in the cyst cavity. No viable protoscolices or daughter cysts were observed. CONCLUSIONS: Percutaneous treatment of cystic echinococcosis with ivermectin as a scolicidal agent seems to be effective in this animal model. PMID- 11910349 TI - Heat stress prevents impairment of bile acid transport in endotoxemic rats by a posttranscriptional mechanism. AB - BACKGROUND & AIMS: Endotoxemia leads to reduction of bile acid transporters in the hepatocyte membrane and impaired bile acid transport. Because heat stress ameliorates other sequelae of endotoxemia, studies were performed to determine whether heat stress would correct deficient bile acid transport caused by endotoxin. METHODS: Body temperature of rats was elevated to 42 degrees C for 10 minutes. Lipopolysaccharide was injected after different time intervals, and maximal transport for cholyltaurine was measured in perfused rat livers. Sodium dependent and -independent uptake was studied in isolated hepatocytes. Protein expression, messenger RNA levels, and tissue distribution of the bile acid transporters sodium taurocholate cotransporting protein (ntcp) and bile salt export pump (bsep) were also analyzed. RESULTS: In the perfused liver, cholyltaurine transport was reduced by 59% by endotoxin, but transport was not reduced when heat stress was applied 2 hours before injection of lipopolysaccharide. The protective effect coincided with maximal expression of heat shock proteins 70 and 25. Sodium-dependent and -independent transport was preserved by heat stress. Expression of bile acid transporters in plasma membrane fractions was reduced after injection of lipopolysaccharide but not if lipopolysaccharide was preceded by heat stress. In contrast, messenger RNA levels of bile acid transporters were not preserved by heat stress. CONCLUSIONS: Heat stress preserves bile acid transporters during endotoxemia by a posttranscriptional mechanism. PMID- 11910350 TI - Neutrophils and NADPH oxidase mediate intrapancreatic trypsin activation in murine experimental acute pancreatitis. AB - BACKGROUND & AIMS: Intrapancreatic activation of digestive enzymes is a key event in the parenchymal cell injury of pancreatitis. We hypothesized that neutrophils recruited to the pancreas during pancreatitis may contribute to such activation. METHODS: To cause experimental pancreatitis, rats and mice were treated with high doses of cerulein. Activation of the digestive enzyme, trypsin, was measured in pancreatic homogenates using a fluorogenic assay and localized immunocytochemically with antibody to trypsin-activation peptide (TAP). RESULTS: Compared with controls, rats depleted of neutrophils with antineutrophil serum exhibited a marked attenuation in intrapancreatic trypsin activation and acinar cell TAP labeling induced by high-dose cerulein. To examine the mechanism, mice deficient in either nicontinamide adenine dinucleotide phosphate (NADPH) oxidase, or myeloperoxidase (MPO) were studied for trypsin activation. Mice deficient in NADPH oxidase exhibited attenuation of the cerulein-induced trypsin activation, but those deficient in MPO did not. Using measurements of Western blot analysis, generation of reactive oxygen species, and immunocytochemistry, we demonstrated the NADPH oxidase activity is in neutrophils and not pancreatic acinar tissue. CONCLUSIONS: The results demonstrate a novel role for neutrophils infiltrating the pancreas in pathologic activation of digestive enzymes in acute pancreatitis and indicate that this effect is mediated by products of NADPH oxidase. PMID- 11910351 TI - Bile acids induce cyclooxygenase-2 expression via the epidermal growth factor receptor in a human cholangiocarcinoma cell line. AB - BACKGROUND & AIMS: Although bile acids have been implicated in colon cancer development, their role in biliary tract carcinogenesis remains unexplored. Because receptor tyrosine kinases and cyclooxygenase (COX)-2 have been implicated in carcinogenesis, we examined the hypothesis that bile acids modulate these enzymes in KMBC cells, a human cholangiocarcinoma cell line. METHODS: The effect of bile acids on epidermal growth factor receptor (EGFR) stimulation, mitogen activated protein kinase (MAPK) activation, and COX-2 expression was evaluated. RESULTS: Bile acids both induced EGFR phosphorylation and enhanced COX-2 protein expression. Bile acid-induced EGFR phosphorylation was associated with subsequent activation of MAPK p42/44, p38, and c-Jun-N-terminal kinase (JNK). The MAPK inhibitors, PD098059 for MAP or extracellular signal-regulated kinase 1, SB203580 for p38, and BAY 37-9751 for Raf-1, blocked COX-2 induction by bile acids. However, inhibition of JNK activity did not block bile acid-mediated COX-2 induction. CONCLUSIONS: The results show that EGFR is activated by bile acids and functions to induce COX-2 expression by an MAPK cascade. This induction of COX-2 may participate in the genesis and progression of cholangiocarcinomas. PMID- 11910352 TI - SNAP-25, a SNARE protein, inhibits two types of K channels in esophageal smooth muscle. AB - BACKGROUND & AIMS: The plasma membrane-associated soluble N-ethylmaleimide sensitive factors attachment protein receptors (SNAREs), synaptosome-associated protein of 25 kilodaltons (SNAP-25), and syntaxin 1A, have been found to physically interact with and functionally modify membrane-spanning ion channels. Studies were performed in cat esophageal body and lower esophageal sphincter (LES) smooth muscle to (1) show the presence of SNAP-25, and (2) determine whether SNAP-25 affects K+ channel activity. METHODS: Single circular muscle cells from the esophageal body and sphincter were studied. Cellular localization of SNAP-25 and K+ channel activity were assessed. RESULTS: SNAP-25 was found in the plasma membrane of all regions examined. Outward K+ currents in body circular muscle were mainly composed of large conductance Ca2+-activated channel currents (K(Ca), 40.1%) and delayed rectifier K+ channel currents (K(V), 54.2%). Microinjection of SNAP-25 into muscle cells caused a dose-dependent inhibition of both outward K+ currents, maximal 44% at 10(-8) mol/L. Cleavage of endogenous SNAP-25 by dialyzing botulinum neurotoxin A into the cell interior resulted in a 35% increase in outward currents. CONCLUSIONS: SNAP-25 protein is present in esophageal smooth muscle cells, and inhibits both K(V) and K(Ca) currents in circular muscle cells. The findings suggest a role for SNAP-25 in regulation of esophageal muscle cell excitability and contractility, and point to potential new targets for treatment of esophageal motor disorders. PMID- 11910353 TI - Facilitation and attenuation of a visceral nociceptive reflex from the rostroventral medulla in the rat. AB - BACKGROUND & AIMS: Noxious inputs from somatic tissue are subject to biphasic descending modulation from the rostroventral medulla (RVM). In the present study, we investigated descending facilitatory and inhibitory influences from the RVM on a visceral nociceptive reflex. METHODS: The visceromotor response (VMR), a contraction of peritoneal musculature during noxious colorectal distention (80 mm Hg, 20 seconds), was quantified as the integrated electromyogram. RESULTS: At 22 sites in the RVM, electrical stimulation produced biphasic effects, facilitating the VMR at low (5, 10, and 25 microA) and inhibiting it at greater (>50 microA) intensities of stimulation. Electrical stimulation at all intensities tested (5 200 microA) in other sites in the RVM only inhibited (30 sites) or only facilitated (12 sites) the VMR to colorectal distention. Activation of glutamatergic receptors in the RVM replicated the effects of electrical stimulation. Reversible blockage (intraspinal lidocaine injection) or irreversible transection of spinal funiculi revealed that descending facilitatory influences from the RVM were conveyed in the ventrolateral/ventral funiculus, whereas descending inhibitory influences were contained in the dorsolateral funiculi. CONCLUSIONS: Spinal visceral nociceptive reflexes are subject to facilitatory modulation from the RVM, providing the basis for a mechanism by which visceral sensations can be enhanced from supraspinal sites. PMID- 11910354 TI - Interferon-alpha activates multiple STAT signals and down-regulates c-Met in primary human hepatocytes. AB - BACKGROUND & AIMS: Interferon (IFN)-alpha therapy is currently the primary choice for viral hepatitis and a promising treatment for hepatocellular carcinoma (HCC). Primary mouse and rat hepatocytes respond poorly to IFN-alpha stimulation. Thus, it is very important to examine the IFN-alpha signal pathway in primary human hepatocytes. METHODS: The IFN-alpha-activated signals and genes in primary human hepatocytes and hepatoma cells were examined by Western blotting and microarray analyses. RESULTS: Primary human hepatocytes respond very well to IFN-alpha stimulation as shown by activation of multiple signal transducer and activator of transcription factor (STAT) 1, 2, 3, 5, and multiple genes. The differential response to IFN-alpha stimulation in primary human and mouse hepatocytes may be caused by expression of predominant functional IFN-alpha receptor 2c (IFNAR2c) in primary human hepatocytes vs. expression of predominant inhibitory IFNAR2a in mouse hepatocytes. Microarray analyses of primary human hepatocytes show that IFN alpha up-regulates about 44 genes by over 2-fold and down-regulates about 9 genes by 50%. The up-regulated genes include a variety of antiviral and tumor suppressors/proapoptotic genes. The down-regulated genes include c-myc and c-Met, the hepatocyte growth factor (HGF) receptor. Down-regulation of c-Met is caused by IFN-alpha suppression of the c-Met promoter through down-regulation of Sp1 binding and results in attenuation of HGF-induced signals and cell proliferation. CONCLUSIONS: IFN-alpha directly targets human hepatocytes, followed by activation of multiple STATs and regulation of a wide variety of genes, which may contribute to the antiviral and antitumor activities of IFN-alpha in human liver. PMID- 11910355 TI - Proteinases and proteinase-activated receptor 2: a possible role to promote visceral hyperalgesia in rats. AB - BACKGROUND & AIMS: PAR-2s are highly expressed throughout the gastrointestinal tract. These receptors are cleaved by trypsin and mast cell tryptase and can be activated by peptides corresponding to the tethered ligand of the receptor (SLIGRL-NH2 for rat). The aim of this study was to determine whether colonic administration of PAR-2 agonists affects visceral sensitivity to rectal distention in conscious rats. METHODS: Abdominal contractions (a criteria of visceral pain) were recorded in rats equipped with intramuscular electrodes. Rectal distention was performed at various times after intracolonic infusion of SLIGRL-NH2 and trypsin. Inflammation parameters and permeability were followed in the colon after the intracolonic injections. Fos expression at a spinal level (L4 L6) was also studied 2 hours after intracolonic injection of SLIGRL-NH2. RESULTS: Rectal distention significantly increased abdominal contractions starting at the RD volume of 0.8 mL. Intracolonic injection of SLIGRL-NH2 (200 microg/rat) and trypsin (200 U/rat), but not vehicle, LRGILS-NH2 (control peptide), boiled trypsin, or SLIGRL-NH2 injected IP, significantly increased (P < 0.05) abdominal contractions for high volumes of distention, 10- and 24-hour postinfusion. SLIGRL NH2-induced hyperalgesia was inhibited by a NK1 receptor antagonist (SR 140333) but not by indomethacin. Intracolonic injection of SLIGRL-NH2 elevated spinal Fos expression and caused increased intestinal permeability but did not cause detectable inflammation. CONCLUSIONS: Intracolonic infusion of subinflammatory doses of PAR-2 agonists activated spinal afferent neurons and produced a delayed rectal hyperalgesia that involves changes in intestinal permeability and the activation of NK1 receptors. These results identify a possible role for proteinases and PAR-2 in the genesis of visceral hyperalgesia. PMID- 11910356 TI - Clostridium difficile toxin A triggers human colonocyte IL-8 release via mitochondrial oxygen radical generation. AB - BACKGROUND & AIMS: Clostridium difficile toxin A causes mitochondrial dysfunction resulting in generation of oxygen radicals and adenosine triphosphate (ATP) depletion. We investigated whether mitochondrial dysfunction is involved in nuclear factor kappaB (NF-kappaB) activation and interleukin (IL)-8 release from toxin A-exposed enterocytes. METHODS: NF-kappaB activation and IL-8 release in response to toxin A were correlated with reactive oxygen intermediate (ROI) generation and ATP production in HT-29 monolayers or HT-29 cells exposed to ethidium bromide (EB) to inhibit mitochondrial function. RESULTS: HT-29 cells exposed to EB showed damaged mitochondria and diminished resting levels of ATP. ROI production in EB-treated cells exposed to toxin A for 30 minutes was significantly reduced. Exposure of wild-type HT-29 cells to toxin A resulted in increased oxygen radical generation and IL-8 production (P < 0.01 vs. control) that was inhibited by antioxidant pretreatment. Degradation of IkappaB was observed within 30 minutes of toxin exposure, before ras homologue (Rho) glucosylation, and was followed by nuclear translocation of NF-kappaB. Toxin A did not increase IL-8 levels in EB-treated cells, whereas IL-8 release in response to IL-1beta was not affected. CONCLUSIONS: Our data support an early role for mitochondria-derived ROIs in stimulation of IL-8 release from colonocytes by toxin A. ROI generation is independent of Rho inactivation and involves nuclear translocation of NF-kappaB before release of IL-8. PMID- 11910357 TI - Corticosteroids modulate the secretory processes of the rat intrahepatic biliary epithelium. AB - BACKGROUND & AIMS: We investigated the expression of glucocorticoid receptors (GcRs) in the intrahepatic biliary epithelium and the role of corticosteroids in the regulation of cholangiocyte secretion. METHODS: GcR was studied by immunohistochemistry, reverse-transcription polymerase chain reaction, and Western blots. The effects of dexamethasone and budesonide on biliary bicarbonate excretion and H+/HCO3- transport processes were investigated in bile fistula rats, isolated intrahepatic bile duct units (IBDUs), and purified cholangiocytes. RESULTS: GcRs were expressed by rat cholangiocytes. Although acute administration of corticosteroids showed no effect, treatment for 2 days with dexamethasone or budesonide increased (P < 0.05) biliary bicarbonate concentration and secretion, which were blocked by the specific GcR antagonist, RU-486. IBDUs isolated from rats treated with dexamethasone or budesonide showed an increased (P < 0.05) activity of the Na+/H+ exchanger (NHE1 isoform) and Cl-/HCO3- exchanger (AE2 member), which was blocked by RU-486. Protein expression of NHE1 and AE2 and messenger RNA for NH1 but not AE2 were increased (P < 0.05) in isolated cholangiocytes by dexamethasone treatment. CONCLUSIONS: The intrahepatic biliary epithelium expresses GcR and responds to corticosteroids by increasing bicarbonate excretion in bile. This is caused by corticosteroid-induced enhanced activities and protein expression of transport processes driving bicarbonate excretion in the biliary epithelium. PMID- 11910358 TI - Enteroinvasive bacteria alter barrier and transport properties of human intestinal epithelium: role of iNOS and COX-2. AB - BACKGROUND & AIMS: Various invasive pathogens cause diarrhea, but the mechanism(s) are poorly understood. We hypothesized that nitric oxide and prostaglandins might modulate chloride secretory and barrier properties of the infected intestinal epithelium and that diarrhea is caused, in part, by altered expression of inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2). METHODS: Studies were conducted in human intestinal epithelial cell lines (HT29/cl.19A, Caco-2, and T84). Cells were infected with enteroinvasive Escherichia coli (EIEC 029:NM) or Salmonella dublin (SD), or nonpathogenic, noninvasive bacteria (Streptococcus thermophilus [ST] and Lactobacillus acidophilus [LA]). Infected cells and controls were tested for transepithelial resistance, chloride secretion, prostaglandin E2, guanosine 3',5'-cyclic monophosphate and adenosine 3',5'-cyclic monophosphate, and protein expression. RESULTS: Cells infected with EIEC or SD, but not uninfected controls or ST/LA-exposed monolayers, showed a progressive reduction in transepithelial resistance starting at 6-12 hours. Infected HT29/cl.19A and Caco-2 cells, but not T84 cells, also showed an increase in total nitrite. Expression of iNOS, and consequently COX-2, was also increased, followed by increased production of prostaglandins and cyclic nucleotides. Furthermore, basal and stimulated chloride secretory responses to various agonists were enhanced in HT29/cl.19A and Caco-2 cells after infection with enteroinvasive bacteria, and this effect was reversed for some agonists by iNOS or COX-2 inhibitors. Increased expression of cystic fibrosis transmembrane conductance regulator and NKCC1 was also observed in EIEC or SD-infected cells vs. controls, secondary to NO synthase activity. CONCLUSIONS: Up-regulation of iNOS and COX-2 by enteroinvasive bacteria can modulate chloride secretion and barrier function in intestinal epithelial cells. Thus, these enzymes represent possible therapeutic targets in infectious diarrhea. PMID- 11910359 TI - The type II inositol 1,4,5-trisphosphate receptor can trigger Ca2+ waves in rat hepatocytes. AB - BACKGROUND & AIMS: Ca2+ regulates cell functions through signaling patterns such as Ca2+ oscillations and Ca2+ waves. The type I inositol 1,4,5-trisphosphate receptor is thought to support Ca2+ oscillations, whereas the type III inositol 1,4,5-trisphosphate receptor is thought to initiate Ca2+ waves. The role of the type II inositol 1,4,5-trisphosphate receptor is less clear, because it behaves like the type III inositol 1,4,5-trisphosphate receptor at the single-channel level but can support Ca2+ oscillations in intact cells. Because the type II inositol 1,4,5-trisphosphate receptor is the predominant isoform in liver, we examined whether this isoform can trigger Ca2+ waves in hepatocytes. METHODS: The expression and distribution of inositol 1,4,5-trisphosphate receptor isoforms was examined in rat liver by immunoblot and confocal immunofluorescence. The effects of inositol 1,4,5-trisphosphate on Ca2+ signaling were examined in isolated rat hepatocyte couplets by using flash photolysis and time-lapse confocal microscopy. RESULTS: The type II inositol 1,4,5-trisphosphate receptor was concentrated near the canalicular pole in hepatocytes, whereas the type I inositol 1,4,5 trisphosphate receptor was found elsewhere. Stimulation of hepatocytes with vasopressin or directly with inositol 1,4,5-trisphosphate induced Ca2+ waves that began in the canalicular region and then spread to the rest of the cell. Inositol 1,4,5-Trisphosphate-induced Ca2+ signals also increased more rapidly in the canalicular region. Hepatocytes did not express the ryanodine receptor, and cyclic adenosine diphosphate-ribose had no effect on Ca2+ signaling in these cells. CONCLUSIONS: The type II inositol 1,4,5-trisphosphate receptor establishes a pericanalicular trigger zone from which Ca2+ waves originate in hepatocytes. PMID- 11910360 TI - Chemoprevention of esophageal adenocarcinoma by COX-2 inhibitors in an animal model of Barrett's esophagus. AB - BACKGROUND & AIMS: Carcinogenesis in Barrett's esophagus (BE) is associated with an increased expression of cyclooxygenase (COX) 2. However, there has been no direct evidence that inhibition of COX-2 prevents cancer in BE. We studied the effect of MF-Tricyclic, a selective COX-2 inhibitor, on the development of BE and adenocarcinoma in a rat model. METHODS: Four weeks after esophagojejunostomy, 105 Sprague-Dawley rats were randomized to a chow containing MF-Tricyclic or Sulindac, or a placebo. Ninety-six (92%) rats completed the study and were sacrificed at 28 +/- 2 weeks. The animals were assessed for the presence of cancer, tumor volume, BE, degree of inflammation, and COX-2 expression and activity. RESULTS: MF-Tricyclic and Sulindac reduced the relative risk of development of esophageal cancer by 55% (95% confidence interval [CI] = 43%-66%, P < 0.008) and by 79% (95% CI = 68%-87%, P < 0.001), respectively, compared with controls. No significant differences were noted in the risk of esophageal cancer between the MF-Tricyclic and the Sulindac group (P = 0.34). The median tumor volume was not significantly different among the 3 groups (P = 0.081). Moderate to severe degree of inflammation was significantly more common (P = 0.005) in the control compared with the MF-Tricyclic and the Sulindac group; however, the prevalence of BE was not significantly different between groups (P = 0.98). Rats in the control group had higher tissue PGE2 level compared with the MF-Tricyclic and Sulindac groups (P = 0.038). CONCLUSIONS: Selective and nonselective COX-2 inhibitors can inhibit inflammation, COX-2 activity, and development of adenocarcinoma induced by reflux. This provides direct evidence that COX-2 inhibitors may have chemopreventive potential in BE. PMID- 11910361 TI - p16 inactivation by methylation of the CDKN2A promoter occurs early during neoplastic progression in Barrett's esophagus. AB - BACKGROUND & AIMS: The potential role of p16 inactivation by CDKN2A/p16 promoter hypermethylation and/or loss of heterozygosity (LOH) of the CDKN2A gene was investigated in neoplastic progression of Barrett's esophagus. METHODS: CDKN2A promoter hypermethylation was studied by methylation sensitive single-strand conformation analysis and sequencing using bisulfite modified DNA in Barrett's esophageal adenocarcinomas, premalignant lesions, and normal squamous esophageal epithelium. All of the lesions of interest were sampled by microdissection from paraffin-embedded fixed tissue sections. RESULTS: No methylation of the CDKN2A promoter was found in normal esophageal squamous cell epithelia, whereas methylation was detected in 18 of 22 (82%) adenocarcinomas and 10 of 33 (30%) premalignant lesions, including 4 of 12 (33%) samples with intestinal metaplasia only. LOH at the CDKN2A gene locus was found in 68% of adenocarcinomas and in 55% of premalignant lesions. Of 28 samples without p16 immunoreactivity, 25 (89%) showed CDKN2A promoter hypermethylation with or without LOH of CDKN2A. Only 2 (8%) samples expressing p16 protein were found to be methylated; these showed a mixture of completely methylated and unmethylated CDKN2A promoters. In 7 of 19 (37%) informative samples without LOH of CDKN2A, the CDKN2A promoter was found to be methylated at both alleles. Loss of p16 protein expression was strongly associated with CDKN2A promoter hypermethylation (P < 0.00001), but not with LOH (P = 0.33). CONCLUSIONS: Our results indicate that methylation of the CDKN2A promoter is the predominant mechanism for p16 inactivation. This hypermethylation is a very common event in esophageal adenocarcinoma and occurs as early as metaplasia. PMID- 11910362 TI - Leukocyte recruitment in colon cancer: role of cell adhesion molecules, nitric oxide, and transforming growth factor beta1. AB - BACKGROUND & AIMS: A deficient leukocyte recruitment has been suggested in tumor vasculature, but little is known about the underlying molecular mechanism. To characterize leukocyte-endothelium interaction in experimental colon cancer, quantify the main endothelial cell adhesion molecules (CAMs), and evaluate the effect of tumor-derived products. METHODS: Leukocyte recruitment was assessed by intravital videomicroscopy in mice bearing HT29-derived tumors. Endothelial CAMs were measured using the dual-radiolabeled antibody technique. The role of molecules mediating leukocyte rolling (P-, E-, and L-selectin) or adhesion (intercellular adhesion molecule 1 [ICAM-1] and vascular cell adhesion molecule 1 [VCAM-1]) carcinoembryonic antigen (CEA), and transforming growth factor (TGF) beta1 was assessed through immunoblockade, whereas participation of nitric oxide (NO) and cyclooxygenase (COX) metabolites were evaluated by means of nonselective and selective inhibition. RESULTS: Basal and lipopolysaccharide-stimulated leukocyte rolling and adhesion were markedly reduced in tumor vasculature. ICAM-1 immunoblockade prevented leukocyte adhesion in both tumor and nontumor microvessels. Neither baseline nor LPS-induced endothelial ICAM-1, P-, and E selectin expression in tumors were reduced with respect to nontumor vasculature. Although VCAM-1 expression was reduced in tumor endothelium, immunoneutralization of VCAM-1 failed to reverse LPS-induced leukocyte recruitment in this setting. CEA immunoblockade and COX inhibition did not modify the deficient leukocyte rolling. Nonselective NO inhibition partially reversed the defective adhesion response in tumor microvessels. Finally, TGF-beta1 immunoblockade partially and selectively restored impaired leukocyte rolling and adhesion in tumor microvessels. CONCLUSIONS: Impaired leukocyte recruitment in tumor vasculature cannot be attributed to a depressed expression of the main CAMs. Selective restoration after NO inhibition and TGF-beta1 immunoblockade suggests involvement of both molecules in this phenomenon. PMID- 11910363 TI - Acquired myopathic intestinal pseudo-obstruction may be due to autoimmune enteric leiomyositis. AB - We describe a previously healthy boy who developed intestinal pseudo-obstruction following an episode of gastroenteritis at age 2 years. At presentation, the patient had mildly raised erythrocyte sedimentation rate and C-reactive protein level, and elevated antineutrophil cytoplasmic antibodies, antinuclear anti-DNA, and anti-smooth muscle antibodies. His electrogastrography was myopathic with no dominant frequency. First full-thickness intestinal biopsies showed a T lymphocytic myositis, particularly in the circular muscle. Steroid therapy resulted in clinical remission; cessation of steroids, in relapse. Further full thickness biopsies showed an initial reduction in alpha-smooth muscle actin immunostaining in circular muscle myocytes and later atrophy and disappearance of many myocytes. Vascular and the remaining enteric smooth muscle cells showed HLA DR and intercellular adhesion molecule 1 expression. These observations demonstrate the ability of enteric myocytes to take part in an inflammatory response and to change their phenotype, allowing them to act as antigen presenting cells and to activate T cells. This and possible cytokine production by the myocytes play a role in their own destruction. This process responded to immunosuppressive therapy. PMID- 11910364 TI - Systematic review of the comorbidity of irritable bowel syndrome with other disorders: what are the causes and implications? AB - BACKGROUND & AIMS: Comorbid or extraintestinal symptoms occur frequently with irritable bowel syndrome and account for up to three fourths of excess health care visits. This challenges the assumption that irritable bowel is a distinct disorder. The aims of this study were to (1) assess comorbidity in 3 areas: gastrointestinal disorders, psychiatric disorders, and nongastrointestinal somatic disorders; and (2) evaluate explanatory hypotheses. METHODS: The scientific literature since 1966 in all languages cited in Medline was systematically reviewed. RESULTS: Comorbidity with other functional gastrointestinal disorders is high and may be caused by shared pathophysiological mechanisms such as visceral hypersensitivity. Psychiatric disorders, especially major depression, anxiety, and somatoform disorders, occur in up to 94%. The nongastrointestinal nonpsychiatric disorders with the best-documented association are fibromyalgia (median of 49% have IBS), chronic fatigue syndrome (51%), temporomandibular joint disorder (64%), and chronic pelvic pain (50%). CONCLUSIONS: Multivariate statistical analyses suggest that these are distinct disorders and not manifestations of a common somatization disorder, but their strong comorbidity suggests a common feature important to their expression, which is most likely psychological. Some models explain the comorbidity of irritable bowel with other disorders by suggesting that each disorder is the manifestation of varying combinations of interacting physiological and psychological factors. An alternative hypothesis is that the irritable bowel diagnosis is applied to a heterogeneous group of patients, some of whom have a predominantly psychological etiology, whereas others have a predominantly biological etiology, and that the presence of multiple comorbid disorders is a marker for psychological influences on etiology. PMID- 11910365 TI - Validity of randomized clinical trials in gastroenterology from 1964-2000. AB - BACKGROUND & AIMS: The internal validity of clinical trials depends on the adequacy of the reported methodological quality. We assessed the methodological quality of all 383 randomized clinical trials published in GASTROENTEROLOGY as original articles from 1964 to 2000. METHODS: The methodological quality (randomization and blinding), sample size, publication year, and disease area were extracted from each trial. Changes during the study period were analyzed by analysis of variance with adjustments for potential confounders. RESULTS: Forty two percent of all trials reported adequate generation of the allocation sequence, 39% reported adequate allocation concealment, and 62% were double blind. The reported methodological quality improved significantly in the mid 1990s. CONCLUSIONS: The present study shows a positive development, but the reported methodological quality of trials can still be improved. PMID- 11910366 TI - Genomics and phenomics in Crohn's disease. PMID- 11910367 TI - Patient and graft survival following liver transplantation for hepatitis C: much ado about something. PMID- 11910368 TI - Failure to yield: drug resistance in inflammatory bowel disease. PMID- 11910369 TI - Pancreatitis severity: who calls the shots? PMID- 11910370 TI - Barrett's-related esophageal cancer: has the final hurdle been cleared, now paving the way for human chemoprevention trials? PMID- 11910371 TI - Acute HCV: the early bird catches the virus. PMID- 11910372 TI - Awakening the sleeping postsurgical abdomen. PMID- 11910373 TI - Cost analysis of antireflux surgery versus medical management. PMID- 11910374 TI - Health-related quality of life after ileal pouch anal anastomosis for ulcerative colitis: right answer-wrong question. PMID- 11910378 TI - Discussion on esophageal motility in reflux disease before and after fundoplication: a prospective, randomized, clinical, and manometric study. PMID- 11910379 TI - ICC in neurotransmission: hard to swallow a lack of involvement. PMID- 11910380 TI - Cost utility of initial medical management for Crohn's disease perianal fistula. PMID- 11910381 TI - Helicobacter pylori infection in childhood: transmission and role of antibiotics. PMID- 11910382 TI - Increasing nursing research in cystic fibrosis. AB - A pressing need exists for more research on the clinical, psychosocial, and genetic dimensions of cystic fibrosis (CF)(a broad spectrum of issues to which nursing research is well suited). The National Institute of Nursing Research (NINR) is therefore seeking ways both to bring new nurse researchers into ongoing CF research and also to encourage those already engaged in nursing research in other areas to widen their focus to include CF. These goals were the focus of the NINR Spring Science Workgroup held on May 1-2, 2001, in Bethesda, Maryland. The NINR brought together an interdisciplinary workgroup of clinical scientists and practitioners in the fields of pediatric and adult cystic fibrosis as well as investigators with research expertise in telehealth, genetics, child development and family interactions, and ethics. The workgroup, chaired by Dr Patricia Grady and Dr Hilary Sigmon, was charged with identifying gaps in current knowledge and suggesting ways to increase the number of nurse researchers engaged in nursing research on CF. PMID- 11910383 TI - Correlates of health-related quality of life in patients with heart failure. AB - BACKGROUND: The purpose of this study was to explore health-related quality of life (HRQOL) in patients with advanced heart failure undergoing heart transplantation evaluation. The overall aim of the study was to determine whether patients' demographic characteristics, functional status, neuroticism, social network, social support, spirituality, and time since symptom onset are related to the physical and mental components of a patient's HRQOL. METHODS: A descriptive, correlational design was used. Patients (N = 61) were recruited from 2 university-affiliated, outpatient, heart failure programs. Data were collected from chart review, a 6-minute walk, and patient-completed instruments. HRQOL, including physical and mental health components, was assessed with use of the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36). RESULTS: Demographic characteristics, functional status, neuroticism, social network, social support, spirituality, and time since symptom onset explained 26% of the variability in the physical health component of HRQOL and 44% of the variability in the mental health component of HRQOL in patients with advanced heart failure. In analyzing the data for the most parsimonious model, New York Heart Association classification, 6-minute walk distance, and neuroticism explained 49% of the variability in the mental health component of HRQOL. CONCLUSIONS: New York Heart Association classification, 6-minute walk distance, and neuroticism are related to the mental health component of HRQOL and can be easily included in the assessment of patients with heart failure who are undergoing heart transplantation evaluation. The findings of the current study require replication but may be used to identify patients with heart failure who are potentially at risk for reduced HRQOL. PMID- 11910384 TI - Health-related quality of life and sense of coherence among elderly patients with severe chronic heart failure in comparison with healthy controls. AB - OBJECTIVE: To describe health-related quality of life (QoL) and sense of coherence (SOC) in a group of elderly people with moderate to severe chronic heart failure and to make comparisons with a healthy sex- and age-matched control group. METHODS: Patients (n = 94), with a mean age of 81 years, hospitalized for chronic heart failure with New York Heart Association functional classification III to IV were age- and sex-matched to a healthy control group. The instruments used were the 36-Item Short Form Health Survey (SF-36) and SOC. RESULTS: The patients had lower levels of health-related QoL scores (SF-36) but high and similar scores of SOC compared with the controls. There were, however, significant positive correlations between the SOC scores and the emotional dimensions in the SF-36 instrument. CONCLUSION: The findings from this study indicate that old age and severe chronic heart failure were associated with limited functional abilities and impaired health-related QoL but also with internal resources such as SOC. PMID- 11910385 TI - Nurses's knowledge of heart failure education principles. AB - OBJECTIVE: The goal of this study was to determine nurses' knowledge of heart failure (HF) self-management education principles. DESIGN: The study was exploratory and descriptive and included a convenience sample. SETTING: Research took place in a large midwestern health care system that included a university based hospital, community hospitals, and home or palliative care. SUBJECTS: Subjects included 300 nurses who provide care to patients with HF. OUTCOME MEASURE: The outcome measures included overall and topic specific perceptions of basic information important to HF self-management. Topics included diet, fluids or weight, signs or symptoms of worsening condition, medications, and exercise. INTERVENTION: A 20-item, true or false written survey was administered between February 2000 and April 2000. RESULTS: Of the 300 nurses surveyed, 92% were registered nurses and 8% were licensed practical nurses; 38% worked in a large university-based hospital; 44% were employed at 5 community hospitals; and 18% worked in home or hospice-palliative care. Mean HF self-management knowledge score was 15.2 +/- 2.0. Registered nurses scored significantly higher than licensed practical nurses (15.3 vs 14.1; P =.004). Individual questions with overall scores <30% were related to dry or ideal weight in daily weight monitoring (24%), nonsymptomatic, low blood pressure (26%), and short-term dizziness when rising (19%). Individual question scores >30% and < or =75% were related to nonsteroidal anti-inflammatory inhibitor use (49%), potassium-based salt substitute use (52%), rest vs activity (72%), and lean delicatessen meat use in a low sodium diet (75%). In questions with scores <30%, nurses requested more information only 5% to 8% of the time. Overall analysis of variance indicated differences by work experience. HF nurses (primary population) scored higher than critical-care, medical-surgical, or telemetry floor nurses (16.2 +/- 1.7; 15.1 +/ 1.8; and 14.7 +/- 2.0, respectively; P <.001); home care nurses scored higher than hospital or palliative care nurses (15.9 +/- 1.5; 15.1 +/- 2.0; and 14.0 +/- 1.5, respectively; P =.006). CONCLUSION: Nurses may not be properly educated in HF self-management principles and must be provided with the right information so they can improve the quality and amount of information they offer to patients. Nurses who are better prepared to educate patients with HF may be more likely to carry out this nursing function as a part of their daily job role. PMID- 11910386 TI - Failed reperfusion after thrombolytic therapy: recognition and management. AB - BACKGROUND: Failed reperfusion after thrombolysis occurs in as many as 30% of patients with acute myocardial infarction (MI). Furthermore, some patients have incomplete tissue perfusion despite reperfusion of the infarct-related artery. Close assessment of the efficacy of thrombolytic administration in people with evolving acute MI is necessary, particularly with regard to myocardial perfusion status, because some patients may benefit from incremental pharmacologic or invasive reperfusion strategies. PURPOSE AND METHOD: This article reviews a number of strategies to assess infarct-related artery patency and myocardial tissue perfusion. These include coronary angiography, continuous ST-segment monitoring, serial electrocardiography, obtaining serial serum biochemical markers of myocardial necrosis, monitoring for reperfusion arrhythmias, and assessment of changes in chest pain intensity. CONCLUSION: The early detection of failed reperfusion is critical if incremental strategies to enhance myocardial salvage are to be considered. Continuous ST-segment monitoring is a relatively inexpensive, reliable, and accurate tool for assessing real-time myocardial perfusion. PMID- 11910388 TI - Effects of phase I cardiac rehabilitation on anxiety of patients hospitalized for coronary artery bypass graft in Taiwan. AB - OBJECTIVE: To assess the effects of phase I cardiac rehabilitation intervention on anxiety of patients hospitalized for coronary artery bypass graft (CABG) surgery. DESIGN: Prospective, quasi-experimental, random assignment, repeated measurements. SETTING: The Veterans General Hospital Taipei, Taiwan, Republic of China. PATIENTS: Seventy patients were randomly assigned to (1) the phase I cardiac rehabilitation intervention (experimental) group and (2) the nonintervention (comparison) group. Ultimately, 60 subjects were included in the data analyses. OUTCOME MEASURES: Psychological status was evaluated by the state of anxiety scores on the State-Trait Anxiety Inventory. Anxiety scores were measured 3 times: (1) after admission, before the patient underwent CABG surgery; (2) the day before the patient underwent CABG surgery; and (3) the day of discharge from the hospital. INTERVENTION: Individual instruction in progressive exercises and daily activities according to the phase I cardiac rehabilitation program (Chinese manual) were used during hospitalization. RESULTS: Data analysis was performed with use of generalized estimating equations (GEE) to assess the between- and within-group variations. The mean anxiety for all subjects before undergoing CABG surgery was 42.6. The mean anxiety on the day before undergoing CABG surgery was 33.7 in the experimental group and 49.8 in the comparison group; there were statistical differences, with a P <.05 level of significance between these 2 groups. The mean anxiety on the day of discharge in the experimental group was 28.6 and in the comparison group was 38.4; there were statistical differences, with a P <.05 level of significance between these 2 groups. CONCLUSIONS: These results have been supported by similar studies. This finding suggests that application of phase I cardiac rehabilitation intervention can reduce the anxiety level during hospitalization of patients undergoing CABG surgery. PMID- 11910387 TI - Preformed anti-human leukocyte antigen antibodies jeopardize cardiac transplantation in patients with a left ventricular assist device. AB - In 1997, 15% of patients who received a cardiac transplant in the United States needed a mechanical circulatory support device before transplantation. One device that patients received was the left ventricular assist device (LVAD). During the LVAD support period, approximately 30% to 80% of LVAD recipients have positive test results for panel reactive antibodies (PRAs). Many of these antibodies form against human leukocyte antigens (HLA). These antigens are present on most cells and stimulate antibody production when a person receives unrelated donor cells. Several pre-LVAD and post-LVAD factors contribute to anti-HLA antibody formation. These antibody levels must be lowered before transplantation because the presence of anti-HLA antibodies makes it more difficult to find a suitable donor and increases the risk of rejection. The objectives of this article are to describe anti-HLA antibody formation in LVAD recipients, review its major consequences and treatments, and discuss nursing actions associated with anti-HLA antibody formation. PMID- 11910389 TI - A pilot study to investigate any relationship between sustained maximal inspiratory pressure and extubation outcome. AB - OBJECTIVE: To establish whether any relationship exists between extubation outcome and sustained maximal inspiratory pressures (SMIP). DESIGN AND SETTING: Prospective clinical study in the 7-bed general intensive care unit of a university hospital. SUBJECTS: Twenty-seven intubated adults who were deemed ready for extubation were enrolled. MEASUREMENTS: Standard respiratory parameters and inspiratory muscle function data (ie, SMIP and peak maximal inspiratory pressures [MIP]) were recorded before extubation. RESULTS: SMIP was found to be significantly greater in those who were successfully extubated than in those who underwent a failed extubation (P <.01). Receiver operating characteristic curves for SMIP data indicated that a cutoff point of 57.5 pressure time units would give a sensitivity and specificity of 1.0 for extubation outcome prediction. Peak MIP was also significantly greater in those successfully extubated (P =.04); a cutoff point of 17.5 cm H(2)O gave a sensitivity of 1.0 and a specificity of 0.5. CONCLUSIONS: In this study, peak MIP was not specific enough to be clinically useful as a predictor of extubation outcome. SMIP was associated with extubation outcome with equally high sensitivity and specificity and may therefore have a role in outcome prediction. PMID- 11910390 TI - Investigation of the difference between treadmill self-efficacy and actual performance in Taiwanese patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: Because overactivity or underactivity may result in inadequate physical responses among patients with chronic obstructive pulmonary disease (COPD), the purpose of this study was to examine the difference between treadmill self-efficacy and actual treadmill performance. Factors that influence self efficacy and actual performance were also examined. DESIGN: The design was a descriptive and correlational study. SETTING: The study took place at the Research Center of Sports Medicine in University. PATIENTS: A total of 48 subjects with COPD were recruited from 4 hospitals. OUTCOME MEASURES: The outcome measures were treadmill self-efficacy and actual treadmill performance. INTERVENTION: Data were collected by means of treadmill exercise testing and 3 structured questionnaires. RESULTS: The findings of the study demonstrated that the average maximal functional capacity was 2.94 METs. A positive significant relationship between treadmill self-efficacy and actual performance was observed. However, the majority of subjects (72.9%) underestimated their treadmill performance and only 7 subjects (14.6%) assessed their treadmill performance accurately. Dyspnea was the most common reason for a subject to stop during the exercise testing. The patient's past experience was the most important predictor for both treadmill self-efficacy and actual treadmill performance. CONCLUSIONS: These results revealed that patients in Taiwan who have COPD have extremely poor functional capacity and most of them underestimated their exercise performance. An assessment of self-efficacy and exercise performance seems imperative in the development of individualized nursing interventions to help COPD patients. PMID- 11910391 TI - Hidden trigeminy and pulsus alternans. PMID- 11910392 TI - Worms take cues from host immune system. PMID- 11910393 TI - The Puppy Mill Protection Act of 2001. PMID- 11910394 TI - Home bred dogs are OK, aren't they? PMID- 11910395 TI - Oozing ostrich egg: omphalitis caused by Enterobacter sp. PMID- 11910396 TI - Conservation and regeneration of transgenic lines of swine by semen cryopreservation and artificial insemination. PMID- 11910397 TI - Pain and distress in research animals: a panel of experts debates the issues. PMID- 11910398 TI - Cage processing and waste management: a cost-analysis and decision-making exercise. PMID- 11910399 TI - Affordable noise control in a laboratory animal facility. PMID- 11910400 TI - Considerations for an automated cage-processing system. PMID- 11910402 TI - Roles and responsibilities of facility owners and attending veterinarians in ensuring adequate veterinary care. PMID- 11910403 TI - Can one vote overrule a majority? In a designated-Member review...maybe. PMID- 11910404 TI - Pink mass on the dorsomedial aspect of a rabbit's eye: cherry eye or prolapse of the deep gland of the nictating membrane. PMID- 11910405 TI - Issues to consider when phenotyping mutant mouse models. PMID- 11910406 TI - Mutant mouse pathology: an exercise in integration. PMID- 11910408 TI - DNA isolation methods for genotyping rodents. PMID- 11910407 TI - Amnesic effects of relative humidity and temperature in mice. PMID- 11910409 TI - Protocol approval: deliberate or just flip of a coin. PMID- 11910410 TI - E-filing of USDA reports; Freedom of Information Act. PMID- 11910411 TI - Did an audiotape squirrel away the minutes? PMID- 11910412 TI - Acute ataxia in a young ferret following canine distemper vaccination. Renal failure after epinephrine overdose. PMID- 11910414 TI - A community-based occupational enrichment program for captive chimpanzees. AB - The authors describe a cost-effective program for providing chimpanzee enrichment that at the same time educates the local community about the care of these animals in research. PMID- 11910415 TI - Developing a rodent enrichment program. AB - The authors describe methods to establish a program for selecting, monitoring, and testing environmental enrichment devices for the rodents in their pharmaceutical facility. They also discuss some of the benefits of such a program for these species that are often overlooked in standard environmental enrichment plans. PMID- 11910416 TI - Environmental enrichment for laboratory marmosets. AB - The authors report on using two environmental enrichment devices for marmosets, and suggest the design of five other devices that may be more successful in stimulating foraging or grooming behavior than the devices tested. PMID- 11910417 TI - Environmental enrichment committee: its role in program development. AB - The authors discuss the role of the Environmental Enrichment Committee in developing, implementing, assessing, and modifying a university animal enrichment program. PMID- 11910418 TI - Group-housing subadult male cynomolgus macaques in a pharmaceutical environment. AB - The authors describe the preliminary results of a program to group-house male cynomolgus monkeys. Using a unique cage design, they were able to achieve environmental enhancement and enrichment that led to easier handling of the animals used in protocols for pharmacological research. PMID- 11910420 TI - [Dental fluorosis in Brazil: a critical review]. AB - This paper discusses dental fluorosis as a relevant public health problem, using a review of epidemiological studies published in the last 10 years on the disease's prevalence, severity, and risk factors. The results suggest that there are already more cases than expected, although few studies refer to major severity. Thus, measures are needed for the prevention and surveillance of dental fluorosis. PMID- 11910421 TI - Chemical exposure during pregnancy and oral clefts in newborns. AB - This article presents a literature review on the risk factors for oral clefts (lip and/or palate), emphasizing discussion of maternal exposure to endocrine disruptors. Several studies have identified the risk of cigarette smoking and alcohol consumption, use of anticonvulsant drugs, and exposure to organic solvents. A protective effect has been shown for supplementation with folic acid. As with other chemicals, the risk associated with exposure to sex hormones is still obscure, although some authors describe a moderate risk level. New studies addressing this hypothesis need to be conducted, while the population exposed to these endocrine disrupters is increasing. PMID- 11910422 TI - [Childhood cancer: a comparative analysis of incidence, mortality, and survival in Goiania (Brazil) and other countries]. AB - Analysis of cancer incidence, mortality, and survival rates can yield geographic and temporal trends that are useful for planning and evaluating health interventions. This article reviews cancer incidence and mortality rates and respective trends around the world in children under 15 years old, as well as their 5-year survival rates in developed and developing countries. We conclude that even though increasing or stable childhood cancer incidence rates and decreasing mortality rates have been observed in developed countries, the trends remain unknown in developing countries. Data from the city of Goiania, Brazil, show stable childhood cancer incidence and mortality rates. Five-year survival rates (48%) in Goiania are similar to those seen in underdeveloped regions and lower than those reported in developed countries (64-70%). PMID- 11910423 TI - Suggestion of an inverse relationship between perception of occupational risks and work-related injuries. AB - Worker perception of risk influences worker behavior and consequently exposure to risks. However, an inverse relationship between perception of occupational risks and work-related injuries has not yet been clearly established. A matched case control was performed aiming to investigate possible differences in perception of occupational risks between workers who had suffered occupational injuries and those who had not. Cases were defined as all 93 workers from a large metallurgical factory in southeastern Brazil, who had suffered occupational injuries during the year 1996. Controls were 372 workers who had not suffered occupational injuries, matched on the basis of the factory sector and jobs performed. Assessment of occupational risk perception was performed by asking the workers to fill out a questionnaire consisting of questions on specific risks related to problems in work relations, work per se, and mode of production. The findings suggest that the degree of perception that workers with occupational injuries have of some occupational risks is lower than that of their non-injured coworkers. PMID- 11910424 TI - [Management of acute respiratory infections in children: evaluation in Rio de Janeiro health care facilities]. AB - The goal of this article was to assess quality of case management for acute respiratory infection (ARI) in children and to detect barriers to proper management. A cross-sectional approach was used with a representative sample of primary and out-patient health care facilities under the Rio de Janeiro Municipal Health Department. Physicians were observed while attending children under five years of age with ARI, and the children were then evaluated according to standard criteria, and the results compared. Physicians were interviewed and health care facilities evaluated for availability of antibiotics. We studied 29 facilities (two hospitals, 20 health centers, and seven health posts), interviewed 46 physicians, and observed 267 children. Sensitivity of the classification used to detect pneumonia was 21.8 (95% CI: 9.3-40.4), specificity was 77.3 (95% CI: 70.3 82.4), and accuracy was 70.6 (95% CI: 64.7-75.5). Antibiotics were prescribed unnecessarily for 8.9% of ARI cases. Standard antibiotics were available in all the health care facilities. We conclude that quality of ARI case management in children can be improved substantially, especially with more training and supervision and better organization of services. PMID- 11910425 TI - [Quality of body mass measurement in Rio de Janeiro public health centers in 1996]. AB - The main purpose of the study was to evaluate the quality of body mass measurement of children under 5 years of age treated in 21 health care units in the city of Rio de Janeiro. A total of 38 health care professionals were observed, 292 children were weighed, and 41 scales were calibrated. Almost all of the mechanical scales were positioned improperly. The procedure of weighing the child "undressed" was not performed in 29.9% of the cases on adult beam scales. Almost all of the scales were calibrated (98%) and showed a high correlation coefficient (0.999). There were no important differences between body mass values obtained by the health care professionals and those read by the observer. Thus, nutritional classification using either value showed a perfect correlation (Kappa correlation coefficient = 1). Although some procedures in obtaining body mass values were unsatisfactory, they did not affect the nutritional classification of children in the present study. PMID- 11910426 TI - [Signs, meanings, and actions associated with Chagas disease]. AB - An anthropological approach was employed to investigate the universe of representations (ways of thinking) and behaviors (ways of acting) associated with Chagas disease in a group of public service workers in Belo Horizonte (both infected and not infected with Trypanosoma cruzi). An attempt was also made to evaluate the repercussions of this universe of representations and behaviors on the lives of the infected individuals. The collection and analysis of the data followed the "systems of signs, meanings, and actions" model developed by Corin et al. (1989, 1992). Sixteen seropositive and 12 seronegative workers were interviewed to compare their ways of thinking and acting towards the disease. Data analysis allowed identification of diverse elements within a context that maximized the limitations imposed by Chagas disease. These should be taken into account in the planning of educational campaigns and elaboration of health care models for Chagas patients. PMID- 11910427 TI - [Quality of life assessment in laryngectomized patients: a systematic review]. AB - Laryngectomy is the main sequela in patients with cancer of the larynx. The authors conducted a systematic review to evaluate the relationship between quality of life and laryngectomy. Ninety-six articles published in scientific journals were identified, and 35 were selected whose main focus was laryngectomy and quality of life in laryngectomized patients. Each article was evaluated by way of systematic review. Evaluation of the qualitative attributes of articles used the Qualitative Assessment Questionnaire (QAQ), a validated and tested instrument. Most articles lacked consistency and methodological rigor in measuring quality of life among laryngectomized patients. The relationship between quality of life and laryngectomy also fell far short of the multidimensional approach of the quality of life construct as suggested by the WHO/QOL (World Health Organization/Quality of Life) group. PMID- 11910428 TI - [Contraceptive methods and adequacy of hormonal oral contraceptive use in the city of Pelotas, Rio Grande do Sul, Brazil: 1992-1999]. AB - A cross-sectional study was conducted to assess contraceptive methods and the adequacy of oral contraceptive use by women aged 20 to 49 years in the city of Pelotas. The results were compared with another cross-sectional study performed in 1992. A sample was randomly selected, including 766 women aged 20 to 49 years. Some 495 of the sample (64.6%) used a contraceptive method, in the following order: oral contraceptives (55.4%), surgical sterilization (22.2%), condoms (10,5%), and IUD (7.7%). Among users of oral contraceptives, 62 (22.2%) had some contraindication. Incorrect use of contraceptive methods was associated with age but not with socioeconomic status. As compared to the previous study, there was a reduction in the use of oral contraceptives. Meanwhile, other methods such as surgical sterilization, condoms, and IUD were used more frequently than in 1992. PMID- 11910429 TI - [Social reproduction of leprosy: a study of patients profile with leprosy in the city of Sao Paulo]. AB - This study discusses the relationship between work and living conditions among leprosy patients enrolled in the Sao Paulo municipal public health system in 1996. Social patterns were studied based on the theory of social determination of the health-disease process. The main purpose of the study was to emphasize evidence of the disease determination network, seeking new knowledge to improve public policies on leprosy. Data were gathered from a sample of leprosy patients registered in the city's public health system. Although patients' families are characterized by a common social thread, different work/life possibilities allow for a classification of patients into three social groups. The majority belong to groups that are marginalized from social production, living in areas where social exclusion is more extreme, on the outskirts of the city. If the trends in this study persist, incident leprosy cases will result from the social exclusion of migrants from Brazil's Southeast and Northeast. The study also discusses the position of young people and female patients in the determination network of this infectious disease in the city of Sao Paulo. PMID- 11910430 TI - [A methodology to evaluate and expand knowledge of adolescents of the elementary school on occupational accidents]. AB - Knowledge concerning work-related hazards and accidents among adolescents enrolled in an elementary public school was evaluated through an analysis of their occupational profile and discussions concerning concepts of risk situations at work and individual and collective measures for accident prevention and control. The results included a significant change in their understanding of work related accidents as identified by statistical tests comparing the proportion of correct answers in a questionnaire applied prior to and at two moments after the intervention. PMID- 11910431 TI - [Characterization of hormone replacement therapy users in Campinas, Sao Paulo]. AB - This study employed a descriptive, cross-sectional, population-based design to characterize climacteric women from Campinas, Sao Paulo State, based on use of hormone replacement therapy (HRT). An area cluster sample was selected with 456 women 45 to 60 years of age, residing in Campinas, based on data from the Brazilian Institute of Statistics and Geography (IBGE). Women were selected by area cluster, and the reference unit was the census tract as defined by the IBGE. Data were collected through home interviews using a structured and pre-tested questionnaire provided by the International Health Foundation/International Menopause Society and by the North American Menopause Society and adapted by the authors. In order to characterize women according to current, past, or no use of HRT, a polytonic logistic regression model was used, with a backward selection process of variables. The authors conclude that the main characteristics of HRT users in the city of Campinas were perimenopausal status and higher literacy and socioeconomic class. PMID- 11910432 TI - [Preventive strategies in the control of dental caries: a research synthesis]. AB - A literature review was conducted on the effectiveness of measures to prevent dental caries. Articles published in scientific journals from 1980 to 1998 and indexed in MEDLINE were selected. Some 210 articles were found and their abstracts classified according to intervention strategies, type of research design, and observed effects. The most frequent preventive measures were the use of fluoride toothpaste (20.5%), fluoride mouthwash (17.2%), and occlusal sealants (18,1%). The main target population was schoolchildren 6-12 years old (53.8%) and teenagers (15.7%). Western Europe produced 58.7% of the articles on this subject, followed by United States (22.7%), Eastern Europe, Asia, Canada, and Latin America. No relationship was identified between program results (effective as opposed to ineffective studies) and the type of preventive practice and research design, indicating that other issues need to be investigated, such as the context of organizational implementation (where, how, and by whom prevention is implemented). Education in oral hygiene as a basic tool and key component in specific preventive action has received little research attention. PMID- 11910434 TI - [Pregnancy in adolescence, associated factors, and perinatal results among low income post-partum women]. AB - This paper compares socioeconomic characteristics, prenatal care, and life styles of three groups of post-partum women, one consisting of adolescents (< 20 years) and the other two of women 20-34 years old, classified according to their history of pregnancy during adolescence. A sample of 3,508 post-partum women was selected from public hospitals in the city of Rio de Janeiro, Brazil, and interviewed just after childbirth. To verify the hypothesis of homogeneity of proportions, chi square tests (chi2) were used. Comparing the three groups, the most adverse conditions were found among the 20-34-year-old mothers with a history of pregnancy during adolescence. These women have the least schooling, the highest rates of smoking and use of illegal drugs during pregnancy, and the fewest prenatal appointments. According to this study, prenatal care proved to be an effective compensatory policy for the prevention of prematurity and low birth weight, especially among adolescent mothers. PMID- 11910433 TI - [The concepts of risk and prevention from the perspective of injecting drug users]. AB - This paper provides an exploratory analysis, using a qualitative approach, to perceptions by injecting drug users (IDUs) on: (a) risks associated with injecting practices; (b) risks of HIV/AIDS; and (c) prevention of HIV/AIDS. The study was conducted in five harm reduction programs in Brazil (the AjUDE-Brasil Project). Forty semi-structured interviews were carried out. The study focuses on the concept of "risk". IDUs were mainly concerned over becoming ill and exposure to overdose and violence. IDUs dealt with individual problems in very specific ways in order to minimize them. They admitted that they were well informed about HIV/AIDS but lacked knowledge on reinfection and other bloodborne diseases. IDUs intended to follow guidelines established by health campaigns, but their addiction usually made it difficult. IDUs also suggested alternative harm reduction strategies and displayed a peculiar reading of concepts concerning sharing their injecting paraphernalia. PMID- 11910435 TI - [Portuguese-language cross-cultural adaptation of the Revised Conflict Tactics Scales (CTS2), an instrument used to identify violence in couples]. AB - This article concerns the evaluation of cross-cultural equivalence between the Revised Conflict Tactics Scales (CTS2) originally developed in English and used to identify violence in couples and a Portuguese-language version for use in Brazil. Besides a broad literature review, evaluation of conceptual and item equivalences involved expert discussion groups focusing on the existence and pertinence of the underlying theoretical concepts and corresponding component items in the Brazilian context. Semantic equivalence involved the following steps: two translations and respective back-translations; an evaluation regarding referential and general (connotative) equivalence between the original instrument and each version; further discussions with experts in order to define the final version; and pre-testing the latter on 774 women. It was possible to establish high-quality conceptual, item, and semantic equivalence between the Portuguese language version and the original CTS2. Acceptability of the version was excellent. Although the results were encouraging, they should be reevaluated in the light of forthcoming psychometric analysis (measurement equivalence). PMID- 11910436 TI - The Bambui Health and Aging Study (BHAS): private health plan and medical care utilization by older adults. AB - The aim of this cross sectional study was to investigate whether holding a private health plan affects the consumption of medical services (hospitalization and visits to a doctor) and use of medications by older adults. All residents in Bambui town (Minas Gerais, Brazil) aged >/= 60 years (n = 1,742) were selected. From these, 92.2% were interviewed and 85.9% were examined (blood tests and physical measurements). After adjustments for worse health status, reported less visits to a doctor, and used a small number of prescribed medications. The main explanation for the aged holding a private health plan was economic, not health. Even though those who had only public health coverage complained more in relation to medical care (70.9%), an important proportion of the aged with a private health care plan presented some kind of complaint (45.2%). Another worrying factor was the difficulty to acquire medication because of financial problems (47.2 and 25.2% reported, respectively). Further investigations are needed to verify whether our results can be generalized to other communities of the country. PMID- 11910438 TI - [Reinventing life: a proposal for a socio-anthropological approach to breast cancer]. AB - This paper analyzes social representations of breast cancer and discusses the possibility of including symbolic aspects in the approach to this disease. The methodology is based on both a critical review of the theme (highlighting socio anthropological issues) and the authors' experience in the field of oncology. The paper thus focuses on: (a) historical aspects of clinical and surgical approaches to breast cancer; (b) theoretical discussion of socially-constructed representations of cancer; and (c) incorporation of the symbolic dimension when dealing with women with breast cancer. The authors conclude that health care policy for women with breast cancer should consider both improvements in technical services and the symbolic dimension involved in living with this disease. PMID- 11910439 TI - [Process assessment of health care: Adequacy of the diabetes mellitus treatment in Pelotas, Southern Brazil]. AB - A study was conducted in 1998 with the aim of assessing adequacy of clinical management of diabetic patients attending public health services in Pelotas, Brazil. Patients were interviewed to gather data on medical consultation, disease, and treatment. Data analysis focused on the "type of treatment" variable with three categories: diet, physical activity, and drug treatment. Seventy-six percent of the patients reported having received nutritional orientation. Only 50% of them reported having followed their diet during the preceding 15 days. Also, 75% of patients reported having received recommendations to perform physical activity. However, only one third had practiced some exercise in the previous month. Among 377 interviewed, 289 (77%) were on medication, and among the patients taking oral hypoglycemic agents many had at least one contraindication. The article discusses the need to enhance physicians' adherence to scientific guidelines for diabetes management. PMID- 11910437 TI - [knowledge level about of American tegumentary leishmaniasis (ATL) and use of alternative therapies in an endemic area in the Amazon Region in the State of Maranhao, Brazil]. AB - The level of knowledge concerning American tegumentary leishmaniasis (ATL) and use of alternative therapies was evaluated in five rural communities in Buriticupu, Maranhao State, Brazil. The study lasted from September 1997 to January 1998. Local inhabitants answered an interview on housing conditions, epidemiological aspects, prevention, standard clinical treatment, and alternative therapies. The study population was 378 (19%) out of a total of 1,980 inhabitants, from Sexta Vicinal (35), Quinta Vicinal (63), Trilha 410 (96), Vila Uniao (85), and Buritizinho (99). Of the interviewees, 72% had little knowledge of ATL transmission, 96% had heard something about the disease from friends, and 60.7% knew ATL by the local term "lesh". Glucantime was the most familiar drug for treatment, while 29.6% referred to use of herbal remedies on the ulcers. Citrus limon (lemon) was the plant most frequently used, and 15.4% of the interviewees used it as a powder spread on the wound. We conclude that the population had little correct knowledge of ATL in the five areas studied, especially with regard to prevention and treatment. PMID- 11910440 TI - [Brazilian policy for the distribution and production of antiretroviral drugs: a privilege or a right?]. AB - This article focuses on the Brazilian National AIDS Program and its policy of distributing and producing antiretroviral drugs, emphasizing links between local decisions and global HIV/AIDS policies. Emphasizing recent developments in the Brazilian and international scenario with regard to access to treatment for people with HIV/AIDS, the article highlights the participation by the pharmaceutical industry, governments, civil society, and UN agencies in establishing responses to the pandemic. The author concludes by identifying transnational activism as a key response to both the power of pharmaceutical corporations and the law of the market (including patent laws), thus fostering global solidarity for people with HIV/AIDS. PMID- 11910441 TI - [A cancer mortality pattern in Brazilian electrical workers]. AB - This study provides the cancer mortality pattern for a cohort of 10,017 electric power plant workers from 1978 to 1994. Standard mortality ratio (SMR) analysis showed lower mortality from all causes (0.47: 0.43-0.51) and from all groups of causes including neoplasms (0.73: 0.60-0.90) among these workers, in comparison with the same sex and age bracket in Rio de Janeiro. In relation to specific cancer sites, standard cancer mortality ratios (SCMRs) greater than one were observed for neoplasms of the small bowel (10.35: 3.34-32.09), gallbladder (2.64: 0.99-7.03), pancreas (1.83: 0.91-3.66), kidney (2.91: 1.39-6.10), and bladder (2.60: 0.58-4.12), as well as for melanoma (1.82: 0.46-7.28). Important limitations were the workers' relatively young age and the short period of time since job admission (less than 15 years). The small number of deaths for each anatomic site led to instability in the study results, with many non-significant SCMRs and wide confidence intervals. Due to these limitations, study results are difficult to interpret in light of currently available evidence. PMID- 11910442 TI - [Individuals and changes in health organizations: a psychosociological approach]. AB - The Brazilian health sector has undergone a severe crisis, affecting the case resolving capacity, efficiency and governability of the health system as a whole and health organizations in particular. Although innovative management systems and tools have been encouraged, such innovations are limited in their ability to spawn organizational change, especially with regard to the challenge of enabling individual adherence to institutional projects and relations involving individuals and organizations. This paper focuses on the French psychosociological approach for analyzing and intervening in organizations, one of whose main thinkers is Eugene Enriquez. In its view of contemporary organizations, this approach focuses on the conflict between reproduction and creation as the main problem to be solved by management processes. While an organization is essentially seen as a place of order and repetition, organizational change implies the challenge of bringing creative individuals into the organization's project, avoiding the trap of controlling their minds and behavior. PMID- 11910443 TI - [Chemical safety, health, and environment: prospects for governance in the Brazilian context]. AB - Chemical safety is acknowledged by Agenda 21 as one of the most serious problems worldwide, involving governance at the national and international levels. In Brazil, chemical safety problems have increased in intensity and extent, far beyond the capacity to deal with them. The problems are all the more serious in Brazil because issues of democracy, security, sustainability, and equity, all fundamental to governance, are still incipient and still far from being solved. New societal arrangements and a new, contextualized and more participatory science form the basis for developing and expanding strategies for governance to deal with the problem of chemical safety. PMID- 11910445 TI - [Labor process and workers' health in charcoal production in Minas Gerais, Brazil]. AB - This study is a tentative approach to the relationship between environmental and occupational health and development in a specific situation: charcoal production in the Jequitinhonha Valley, the poorest region in the State of Minas Gerais, Brazil. The study focuses on the labor process, involving heavy exploitation, hazardous working conditions, and extremely precarious living conditions. Working conditions and activities are described and related to the workers' health situation. Social policies and decisive participation by the various social stakeholders are necessary to change this reality. PMID- 11910444 TI - [Changes in perinatal care as a determinant of the level and diversity of antiinfectives use in a neonatal intensive care unit in Rio de Janeiro, Brazil]. AB - This study focuses on a five-year profile (1990-1994) in the utilization of antiinfectives in a neonatal intensive care unit (NICU) in Rio de Janeiro, Brazil. Total antiinfectives consumption, diversity, and possible determinants were analyzed. A descriptive, longitudinal, and retrospective drug utilization study design was adopted. Document research, observation, and semi-structured interviews provided information on the neonatal intensive care provided, and drug supply and daily hospital records provided data on antiinfectives use, based on the Anatomical Therapeutic Chemical (ATC) classification and the infant defined daily dose (DDDi) as units of measurement. The numerical results suggest an increase in total consumption of antiinfectives and a decrease in their diversity. Analyzed in view of the institutional profile and rational drug use, these results indicate that changes in the type of care provided during the study period were key determinants of antiinfectives use. PMID- 11910446 TI - The estimated magnitude of AIDS in Brazil: a delay correction applied to cases with lost dates. AB - The number of HIV-infected people is an important measure of the magnitude of the AIDS epidemic in Brazil and allows for comparison with epidemic patterns in other countries. This quantity can be estimated from the number of reported AIDS cases, which in turn needs to be corrected for the distribution of reporting delays and under-recording of cases. These distributions are unknown and must also be estimated from the recorded dates, which were missed to the Brazilian National AIDS registry. This paper estimates the number of AIDS cases diagnosed by inputting the lost information based on an estimate of the pattern in registration delay until 1996. We first fitted a non-stationary bivariate Poisson regression model to estimate the pattern in reporting delay. In the subsequent steps these models were applied to input new data, thus replacing the missing information, and to estimate the magnitude of the AIDS epidemic in the country. Model estimates ranged from 36,000 to 50,000 AIDS cases diagnosed in Brazil and still unreported. Therefore, the epidemic was 20 to 30% greater than known from the available information as of February 1999. To be useful to health policy makers, the surveillance system based on officially reported AIDS cases must be continuously improved. PMID- 11910447 TI - [Evaluating the potential of an intervention aimed at promoting oral rehydration therapy (ORT) by educating pharmacy employees]. AB - Diarrhea is a major cause of morbidity and mortality in childhood, and Brazilians rely heavily on pharmacies for the resolution of this and other health problems. To promote the rational use of both pharmaceuticals and oral rehydration therapy (ORT), an intervention study was performed in pharmacies in Southwestern Brazil. Semi-structured interviews showed oral rehydration solution, or ORS (50%), antidiarrheals (39%), trimethoprim-sulfamethoxazole (27%), and yeast (22%) to be the most frequently suggested drugs, whereas questionnaire responses were ORS (75%), trimethoprim-sulfamethoxazole (25%), and yeast (25%), thus revealing that more than one methodology is needed if reliable data are to be obtained. An educational intervention was applied to 86.7% of pharmacies, but acquisition of knowledge on management of diarrhea did not prevent pharmacy workers from suggesting antidiarrheal drugs instead of ORS alone. In order to have pharmacy workers comply with official protocols for episodes of diarrhea, interventions should include regulatory measures on drugs that are contraindicated for children, and the role of pharmacies and pharmacists should be reviewed. PMID- 11910448 TI - [Use of health service to children under five years of age in Southern Brazil]. AB - To identify the main determinants of health services utilization by children under five years of age, a population-based study using systematic sampling was conducted in Rio Grande, Southern Brazil. Twelve previously trained interviewers applied a standardized home questionnaire to mothers in order to obtain data on living conditions, medical consultations, and hospitalization for children under five years of age. Among 514 children covered by the study, 50% had required consultation with a physician in the previous three months and 11% had been hospitalized in the previous twelve months. Acute respiratory infection was responsible for almost two-thirds of the consultations and half of the hospitalizations. After adjusting the analysis for several confounders, the most important determinants were children's age, father's schooling, and type of home construction. Identification of these factors can contribute to adequate planning of future health interventions and to reach children in the community who need but have not received health care. PMID- 11910449 TI - [Diffusion of science, communication, and health]. AB - The diffusion of science has been identified as an instrument and even as a social movement capable of helping strengthen citizenship and improving the health of populations. The article expounds on the integration of aspects involved in the diffusion of science based on science itself, education, language, and communication, as well as an understanding and social control of science. The article discusses the role of those who produce knowledge in health science as well as those who disseminate it, focusing on such loci as health institutions and schools. The link between various fields of knowledge and practice highlight possibilities and obstacles in the historical and social context. PMID- 11910450 TI - [Health professionals' reporting of family violence against children and adolescents]. AB - Reporting family violence against children has two main benefits: it protects the child from the violence and improves epidemiological control of violence. Health professionals play an important role in this area, since they are required to report any known or even suspected case of violence. Nevertheless, when and how to report has been questioned recently. This paper discusses the problems faced by health professionals and suggests specific solutions to the Brazilian case. The authors conclude that it is necessary: (a) to clarify the legal notion of violence in Brazil, specifically the concept of suspicion; (b) to create technical manuals to guide action in this area; (c) to improve the number and quality of services to assist the population; (d) to improve studies and discussion of the consequences of reporting, mainly concerning the notion of justice transmitted to Brazilian families through this practice. PMID- 11910451 TI - [Tools for planning a project to promote adolescent health and development: the adolescents' perspective]. AB - This qualitative study was developed with low-income teenage students from outlying cities around the Federal District, Brazil. The main objective was to identify their opinions, feelings, and information concerning the community's reality in order to implement a project to promote health care in this age group. A focal group technique was used to collect data. Two focal groups of adolescents 13 to 17 years old were conducted. Resulting data were submitted to descriptive analysis. According to the findings, adolescents have limited opportunities to engage in leisure activities. Problems in the community include lack of security, unavoidable contact with violence, and drug abuse. Adolescents understand that such problems are due to the absence of an appropriate social context. They also highlight difficulties in establishing healthy interpersonal relationships within their families. Their first feeling is one of disempowerment in dealing with prevailing conditions, but they also show willingness to become involved in community work. PMID- 11910452 TI - [Medical residence in pediatrics: in the field of practice]. AB - The study focuses on how specificity of pediatric practice at the Fernandes Figueira Institute, Oswaldo Cruz Foundation, contributes to the development of clinical reasoning, the ability to detect evolution in serious illness, and the capacity to use diagnostic and therapeutic techniques. Data were collected using a thematic interview and analyzed by a semiotic model. The results showed a common view of medicine as both science and art, the doctor-patient relationship as legitimating medical knowledge, pediatrics as having its own peculiarities (thus being defined as a medical specialty), and a severity postulate that hinders the development of clinical reasoning and thus the ability to detect evolution in serious illness. PMID- 11910453 TI - Toxicity of tetracyclines and tetracycline degradation products to environmentally relevant bacteria, including selected tetracycline-resistant bacteria. AB - Tetracyclines used in veterinary therapy invariably will find their way as parent compound and degradation products to the agricultural field. Major degradation products formed due to the limited stability of parent tetracyclines (tetracycline, chlortetracycline, and oxytetracycline) in aqueous solution were theoretically identified at various environmental conditions, such as pH, presence of chelating metals, and light. Their potency was assessed on sludge bacteria, tetracycline-sensitive soil bacteria, and tetracycline-resistant strains. Several of the degradation products had potency at the same concentration level as tetracycline, chlortetracycline, and oxytetracycline on both the sludge and the tetracycline-sensitive soil bacteria. Further, both 5a,6 anhydrotetracycline and 5a,6-anhydrochlortetracycline had potency on tetracycline resistant bacteria supporting a mode of action different from that of the parent compounds. PMID- 11910454 TI - Determination of toxicokinetic parameters for bioconcentration of water-soluble fraction of petroleum hydrocarbon associated with no. 0 diesel in Changjiang estuary and Jiaozhou bay: model versus mesocosm experiments. AB - A method is proposed for determination of toxicokinetic parameters for bioconcentration by phytoplankton of the water-soluble fraction (WSF) of petroleum hydrocarbon (PH) associated with No. 0 diesel, in which WSF-PH concentration in phytoplankton cells, C(A(d)), is estimated by subtracting concentration in water (S-bottle) containing a phytoplankton sample from that in a C-bottle without phytoplankton. It was demonstrated that C(A(d)) agrees well with the concentration found by direct ultrasonication extraction of collected cells, C(A(ind)) ( r = 0.88, p < 0.0001), and its uncertainty was about 17.6%. Mesocosms in 25-m3 ethylene vinyl acetate or 4-m3 polyethylene bags were performed at two sites in China: Changjiang Estuary in spring/summer 1998 and Jiaozhou Bay in autumn 1999 and spring/summer 2000. The experiments were designed to determine toxicokinetic parameters, including specific rates of uptake and elimination, and bioconcentration factor (BCF), for bioconcentration of WSF-PH by phytoplankton. A modified kinetic two-compartment model for bioconcentration of WSF-PH by phytoplankton was developed to estimate the toxicokinetic parameters. In the model, the influence of phytoplankton growth on bioconcentration and WSF PH decline due to biotic and abiotic processes other than bioconcentration, such as volatilization, microbial degradation, phytolysis, and sorption expressed as an exponential-decay equation, are taken into account. Size-dependent BCF was observed in the laboratory experiment. BCFs were 1.0 x 10(4) in summer in Changjiang Estuary, 1.6 x 10(4) in summer, and 1.1 x 10(4) in autumn in Jiaozhou Bay. The difference in BCF may be interpreted by its size dependence. PMID- 11910455 TI - Soil metabolism of isoxaflutole in corn. AB - The herbicide isoxaflutole 1 (5-cyclopropyl-4-isoxazolyl)[2-(methylsulfonyl)-4 (trifluoromethyl)phenyl]-methanone) has been applied preemergence at the rate of 125 g ha(-1) on corn crops grown on fields located in regions different as to their soil textures. Its metabolite diketonitrile 2 (2-cyano-3-cyclopropyl-1-(2 methylsulfonyl-4-trifluoromethylphenyl)propane-1,3-dione)-which is the herbicide's active compound-and its nonherbicide metabolite 3 (2-methylsulfonyl-4 trifluoromethylbenzoic acid) were measured in the 0-10 cm surface soil layer of the corn crops after the treatment and until the harvest. At the opposite of what occurred in plant shoots, the transformation of isoxaflutole 1 into diketonitrile 2 was not immediate in soil. In the 0-10 cm surface soil layer, this transformation occurred progressively according to an apparent second-order kinetics, and the soil half-lives of isoxaflutole 1 self were comprised between 9 and 18 days. The adsorption of isoxaflutole 1 onto the solid phase of the soil and its organic matter should explain the stabilization effect of soil, increased by the application of fresh organic fertilizer. The sum of the concentrations of isoxaflutole 1 and diketonitrile 2 disappeared in the 0-10 cm surface soil layer according to an apparent first-order kinetics, and the soil half-lives of this sum were comprised between 45 and 65 days. The sum of the concentrations of isoxaflutole 1 and of its metabolites diketonitrile 2 and acid 3 did not account for the amount of isoxaflutole 1 applied. The discrepancy increased with the delay after the application, showing that the acid 3 was further metabolized in soil into common nontoxic products, and ultimately into CO2. The conjunction of the adsorption of isoxaflutole and its metabolites (which reduced their mobilities) onto the soil and its organic matter, and their further metabolism should explain why isoxaflutole and its metabolites were not detected in the 10 15 and 15-20 cm surface soil layers during the crops. PMID- 11910456 TI - The potential association between menta deformities and trace elements in Chironomidae (Diptera) taken from a heavy metal contaminated river. AB - Elevated morphological deformity rates in Chironomidae larvae have been attributed to such pollutants as oil tars, pesticides, organochlorines, and heavy metals. The potential of chironomids as biological indicators of heavy metal contamination and bioavailability in a contaminated ecosystem was assessed. Chironomid larvae and river sediment were collected from the Coeur d' Alene River system, Idaho, USA, contaminated with heavy metals from mining activities. Chironomid larvae collected at contaminated sites exhibited mouthpart deformity proportions between 3.8 and 10.3% compared to the control site of 0.9%. As, Cd, Cu, Ni, Pb, and Zn were determined in both larvae and sediment samples. Significant correlation between metal concentrations and deformity rates were observed for all metals except Ni. The data also suggests that feeding habits may influence differences in pollution tolerance among genera. PMID- 11910458 TI - Cellular biomarkers in native and transplanted populations of the mussel Perumytilus purpuratus in the intertidal zones of San Jorge Bay, Antofagasta, Chile. AB - Cellular biomarkers were measured in the mussel Perumytilus purpuratus from intertidal zones of San Jorge Bay, Antofagasta, Chile. They were also used to measure sublethal effects on individuals exposed to Cu under laboratory conditions. Lysosomal stability in hemocytes, and the degree of vacuolization and the content of lipofuscin granules in digestive cells were the cellular responses measured. Three study sites were established in San Jorge Bay: Coloso, E.R., and Reference. Both E.R. and Coloso receive effluent discharges. Reference does not receive any sewage discharges. Before sampling, mussels from Reference were transplanted into the intertidal zone of each site. Samplings were obtained at the beginning, after 45 days, and after 90 days after transplantation. Seawater samples for total dissolved Cu analysis and adult mussels ( P. purpuratus) from native and transplanted populations were collected each time. Cellular biomarkers and Cu concentrations in gonads, gills, and remaining tissues (gut and muscle) were measured. Mussels from Reference were exposed to sublethal Cu concentrations (5, 10, 20, 30, 40, and 80 microg x L(-1)) during 45 days under laboratory conditions. Lysosomal stability was measured in mussel hemocytes by means of the neutral red retention assay. The degree of vacuolization and the extent of lipofuscin granules were determined in the digestive cells by image analysis of histological sections stained with the Schmorl's method. Seawater Cu concentrations and tissue Cu concentrations in P. purpuratus were higher in E.R. than in Reference and Coloso (p < 0.02). Native mussel populations from E.R. showed lower lysosomal stability (p < 0.05), higher vacuolization degree (p < 0.001), and lower amounts of lipofuscin granules (p < 0.001) than those from Coloso and Reference. Transplanted mussel to E.R. showed significant reduction in lysosomal stability (p < 0.05) and in extent of lipofuscin granules (p < 0.05) and significant increase in vacuolization degree (p < 0.05), whereas Reference and Coloso are not significantly dissimilar between them. Seawater Cu concentration was positively correlated with Cu content in gonads ( r2 = 0.61; p < 0.02), gills ( r2 = 0.66; p < 0.01), remaining tissues ( r2 = 0.56; p < 0.05), and the degree of vacuolization ( r2 = 0.65; p < 0.01) and negatively with lysosomal stability ( r2 = 0.79; p < 0.001) and lipofuscin granules extent ( r2 = 0.53; p < 0.05). Mussels exposed to Cu under laboratory conditions showed decreased lysosomal stability (over 30 microg Cu x L(-1)) (p < 0.02) and increased degree of vacuolization (at 80 microg Cu x L(-1)) (p < 0.05) and an increased lipofuscin granules extent (although differences among treatments were not statistically significant). PMID- 11910457 TI - Biological responses of Lumbriculus variegatus exposed to fluoranthene-spiked sediment. AB - Lumbriculus variegatus was used as a bioassay organism to examine the impact of the sediment-associated polycyclic aromatic hydrocarbon (PAH) fluoranthene on behavior, reproduction, and toxicokinetics. The number of worms increased between the beginning and end of the experiment at 59 microg x g(-1) fluoranthene, but at the next higher treatment (108 microg x g(-1)) the number of worms found was lower and not different from the control. Worms exposed to 95 microg x g(-1) also exhibited increased reproduction when fed a yeast-cerophyl-trout chow mixture. On a total biomass basis, only the 95 microg x g(-1) exposure with food exhibited a statistically significant increase over the nonfed control. Evaluation of reproduction at the two highest treatments was compromised by a brief aeration failure 2 days before the end of the experiment. The behavioral responses were followed as changes in biological burial rate (sediment reworking rate) of a 137Cs-labeled marker layer. The biological burial rate increased toward a plateau as the concentration increased from the control (3.9 microg x g(-1) dry weight total PAH) to 355 microg x g(-1) dry weight fluoranthene in sediment. The aeration failure had minimal impact on the determination of reworking rate because all the data for the rate determination were collected prior to the aeration failure. Uptake and elimination rates declined with increasing treatment concentration across the range of fluoranthene concentrations, 59-355 microg x g( 1) dry weight sediment. The disconnect between the increasing biological burial rates and the decreasing toxicokinetics rates with increasing exposure concentration demonstrates that the toxicokinetic processes are dominated by uptake and elimination to interstitial water. The bioaccumulation factor (concentration in the organisms on a wet weight basis divided by the concentration in sediment on a dry weight basis) ranged from 0.92 to 1.88 on day 10 and declined to a range of 0.52 to 0.99 on day 28 with the lowest value at the highest dose. PMID- 11910459 TI - Perfluorooctane sulfonate in oysters, Crassostrea virginica, from the Gulf of Mexico and the Chesapeake Bay, USA. AB - Concentrations of perfluorooctane sulfonate (PFOS), a metabolite of several sulfonated perfluoroorganic compounds, were measured in oysters collected from 77 locations in the Gulf of Mexico and Chesapeake Bay of the United States. PFOS was detected in oysters collected from 51 of the 77 locations at concentrations ranging from < 42 to 1,225 ng/g on a dry weight basis. This study provides baseline data for future monitoring programs to examine long-term trends in concentrations of PFOS. PMID- 11910460 TI - Mercury in Morelet's crocodile eggs from northern Belize. AB - Recent studies have examined mercury accumulation in crocodilians. However, though most researchers have focused on tissue concentrations, few have examined mercury levels in crocodilian eggs. In July 1995, we analyzed mercury in 31 nonviable Morelet's crocodile ( Crocodylus moreletii) eggs collected from eight nests across three localities in northern Belize. All eggs were found to contain mercury. Based on an individual egg basis, mean concentration of mercury for all three localities was among the lowest reported for any crocodilian species. When localities were examined separately, mean concentrations for Laguna Seca and Gold Button Lagoon were comparable to those observed in other studies, and the mean for Sapote Lagoon was the lowest ever reported. Based on mean nest concentrations, mercury in eggs from Laguna Seca was approximately two- and tenfold higher than for Gold Button Lagoon and Sapote Lagoon, respectively. Variability in mercury concentrations among localities is likely the result of site-specific differences in mercury input, bioavailabilty, and bioaccumulation. Mercury concentrations were relatively uniform in eggs from the same nest and among nests from the same localities. The presence of mercury in Morelet's crocodile eggs suggests exposure in adult females, developing embryos, and neonates. However, crocodiles in these areas show no overt signs of mercury toxicity, and no indication of population decline is evident. A paucity of data on the effects of mercury on crocodilians precludes meaningful speculation as to the biological significance of tissue and egg concentrations. Controlled laboratory studies and long-term population monitoring are needed to address these questions. PMID- 11910462 TI - Effects of sublethal levels of tributyltin chloride in a new toxicity test organism: the Chinese rare minnow ( Gobiocypris rarus). AB - A newly developed toxicity test organism, the Chinese rare minnow ( Gobiocypris rarus), which has similar merits to some common experimental fishes and could become a standardized test species for China, was used in a short-term study with emphasis on accumulation in fish muscles and cytological effects of sublethal concentrations of tri-n-butyltin chloride (TBT). The 1-year-old fish were exposed for 1 or 2 weeks to a concentration range of 50 to 5,000 ng TBT/L in static systems. Hepatosomatic Index (HSI) and Gonad Somatic Index (GSI) sensitively showed the adverse effects of TBT to the fish. Compared with the nominal TBT concentrations in water phase, 459-(5,000 ng TBT/L, 1 week) to 4,065- (50 ng TBT/L, 2 weeks)-fold higher concentrations of butyltin species were detected in the corresponding exposure fish muscle. Contaminations of TBT in muscle tissue increased with both the exposure levels and exposure time. The cellular pathological effects in the liver were studied. These included vacuoles with increasing number and size, swelling of mitochondria, abnormal nuclei, and decreases of rough endoplasmic reticulum cisternaes. The results confirmed that the Chinese rare minnow ( G. rarus) could be utilized as a useful species to evaluate water toxicity in the laboratory. PMID- 11910461 TI - Copper speciation and accumulation in the gill microenvironment of carp ( Cyprinus carpio) in the presence of kaolin particles. AB - Carp were exposed to copper adsorbed on kaolin particles at various concentrations (0-2.4 mg/L), and net accumulation due to elevated adsorbed copper in the surrounding water was observed. Copper speciation in the water and fish gill microenvironment was modeled in the presence and absence of kaolin using a chemical speciation program (MINTEQA2). The adsorption affinity constants of kaolin for copper at various pH values used in the speciation calculations were experimentally determined, and the quantitative relationship between the affinity constant and pH was modeled. Copper accumulations in fish gills exposed to kaolin adsorbed copper was then observed. The results indicate that desorption occurred in the fish gill microenvironment due to both mucus competition of copper and slight increase in water pH. Furthermore, the available copper species increased as a result of desorption, causing a net accumulation of Cu by the gills. PMID- 11910463 TI - Platinum group elements in the feathers of raptors and their prey. AB - Platinum (Pt), palladium (Pd), and rhodium (Rh) concentrations were determined in the feathers of three raptor species in Sweden, the sparrowhawk ( Accipiter nisus), the peregrine falcon ( Falco peregrinus), and the gyrfalcon ( Falco rusticolus), as well as the main prey of the sparrowhawk (the house sparrow, Passer domesticus) and the gyrfalcon (the willow grouse, Lagopus lagopus). The analysis of feathers from 1917-1999 revealed a clear temporal trend, with significantly higher Rh concentrations in sparrowhawk and peregrine falcon after 1986. There is evidence for increasing platinum group element (PGE) concentrations from 1917 to 1999 in peregrine falcon and sparrowhawk. This suggests that feathers reflect increased PGE concentrations in the environment over this time period. Mean concentrations of PGE in feathers of raptors after 1986 ranged from 0.3 to 1.8 ng x g(-1) for Pt, 0.6 to 2.1 ng x g(-1) for Pd (indicative values), and 0.1 to 0.6 ng x g(-1) for Rh. House sparrows in urban areas had significantly higher Pt and Pd concentrations than urban sparrowhawks. The higher Pd concentrations in relation to Pt and Rh may indicate the greater mobility of Pd in the environment. Although PGE concentrations are generally higher in birds living in urban areas, no significant spatial trend could be established. This is partly due to the widespread distribution of automobiles and partly because birds forage and integrate PGE exposure over large areas. Laser ablation analysis demonstrates that PGE contamination of feathers is predominantly external, consisting of small particles in the nanometer size range. Other indications of external contamination are that Pt and Pd levels are significantly higher in the vane than in the shaft and that PGE relative ratios (except Pd) reflect urban particles. PMID- 11910464 TI - In vitro study of methylmercury in blood of bottlenose dolphins (Tursiops truncatus). AB - The biochemical behavior of methylmercury (MeHg) in dolphin blood was investigated in vitro. MeHg distribution between plasma and erythrocytes and its release from erythrocytes into plasma or medium without SH group was determined. At the subcellular level its distribution among different thiol-containing molecules was also investigated in erythrocytes and plasma. When blood was treated with 0.1 mM MeHg, about 98.1% was found in red cells and 1.9% in plasma; only 0.6% of MeHg present in the cellular compartment was bound to membranes. Hemoglobin (Hb) and albumin, principal proteins containing SH groups (PSH), and glutathione (GSH) appeared to be the main targets of MeHg in dolphin blood. Gel filtration of stroma-free hemolysate of treated red blood cells (RBCs) revealed that MeHg was almost equally present in high (52.5%) and low (47.5%) molecular weight fractions, whereas in plasma it only eluted with proteins (high molecular weight fractions). Hemoglobin was identified as the main intracellular protein binding MeHg. The exchange reaction of MeHg between GSH and dolphin hemoglobin was also evaluated and the equilibrium constants calculated. PMID- 11910465 TI - Elevated concentrations of trace elements in Caspian seals (Phoca caspica) found stranded during the mass mortality events in 2000. AB - Concentrations of V, Mn, Fe, Cr, Co, Cu, Zn, As, Se, Mo, Ag, Cd, Tl, Hg, Pb, and organic mercury (Org-Hg) were determined in liver, kidney, and muscle of healthy Caspian seals ( Phoca caspica) collected in 1998. These concentrations were compared with those of seals infected with canine distemper virus (CDV) found stranded along the coastal areas in 2000. Concentrations of toxic elements (As, Ag, Cd, Tl, Hg, Pb, and Org-Hg) in Caspian seals stranded in 2000 were comparable or lower than those of samples collected in 1998 and in other pinnipeds. Thus it may be inferred that these elements were not the causative agents in the deaths of the seals. In contrast, concentrations of Zn and Fe were much higher in diseased Caspian seals than those in other pinnipeds. Zinc concentrations in all tissues of Caspian seals also increased during 1993-2000. Furthermore, negative correlations were found between blubber thickness and hepatic and renal Zn concentrations. These results imply the disturbance in homeostatic control and nutritional status of essential elements in Caspian seals stranded in 2000. PMID- 11910466 TI - Content and redistribution of vitamin E in tissues of Wistar rats under oxidative stress induced by hydrazine. AB - Hydrazine toxicity is associated with generation of several kinds of free radicals and oxidative stress in cell. Experiments in vivo have demonstrated that oxidative stress could either diminish or increase concentration of vitamin E in some tissues. Thus in the present study we performed experiments to determine whether hydrazine-induced oxidative stress would change the tissue levels of the vitamin. Seven days of hydrazine intoxication led to accumulation of different amounts of vitamin E: 215% in the liver, 118% in the heart, 135% in the spleen, and 100% in the muscle over control value. There were no changes in the level of the vitamin in kidney and pancreas, despite its significant depletion in the serum. In tissue that accumulated vitamin E after hydrazine treatment, an increased of oxidative stress measured by the concentration of lipid-soluble fluorophore was observed. Significant increases of 107%, 46%, 72%, and 58% over control values were observed in the liver, heart, spleen, and muscle, respectively. Rats treated with hydrazine and pharmacological doses of alpha tocopherol accumulated higher concentrations of vitamin E in all studied tissues compared with the alpha-tocopherol-only treated rats. However, in tissues with elevated levels of fluorophore as liver, heart, spleen, and muscle, the accumulation of vitamin E was 5.03, 4.5, 4.03, and 4.6 times higher than in alpha tocopherol-treated rats, respectively. Vitamin E concentration was much higher than in kidney and pancreas, where the accumulation was only 2.31 and 2.6 times higher. On the other hand, 3 days of hydrazine treatment did not change either the level of lipid-soluble fluorophore or the level of vitamin E in the liver mitochondria, microsomes, and homogenate. In skeletal muscle vitamin E caused decreased lipofuscin accumulation, and in pancreas vitamin E increased lipofuscin accumulation. Our data indicate that hydrazine is able to modify significantly vitamin E status in different rat tissues. PMID- 11910467 TI - Exposure of ground-rig applicators to the herbicide bromoxynil applied as a 1:1 mixture of butyrate and octanoate. AB - Bromoxynil is a herbicide used extensively on the Canadian prairies for weed control in cereal production. This is a report on exposure to and absorption of bromoxynil by farmers during handling, transferring, mixing, and then applying the herbicide to cereal crops using tractor-drawn ground-rigs. The 14 individual spray operations, in which farmers applied 9 kg of bromoxynil (phenol equivalent, p.e.) to 32 ha, lasted from 113 to 549 min and involved one to five tank fills. In five of the spray operations, the farmers wore neoprene gauntlet-style protective gloves. Tractors, equipped with cabs, were used in nine spray operations.Air sampling, hand washes, and dermal patch dosimeters served as the basis to calculate the amount of bromoxynil (p.e.) available for inhalation, deposition on the hands, and deposition under a standardized set of protective clothing. Tractors, equipped with cabs, decreased inhalation exposure by a factor of approximately 10. Use of gauntlet-style neoprene protective gloves decreased dermal exposure to the hands region of the body by 25 times, whereas two layers of laundered cotton provided a protective effect of > 15 in the chest region. The median value for the amount of bromoxynil (p.e.) inhaled was 0.018 microg x kg BW(-1)x kg (p.e.)(-1) for the 14 spray operations. The hands region of the body was the area most exposed to bromoxynil (p.e.), the median value of dermal deposition to the hands being 808 microg (p.e.) compared to 1,600 microg (p.e.) for the whole body, when protective gloves were not worn. The urinary excretion pattern of bromoxynil (p.e.) did not demonstrate an obvious maximum followed by a continuous decline to background concentrations. Instead, the amounts excreted increased over the first few days after application and then tended to remain relatively constant during the remainder of the 10-day sampling period. The median value for urinary excretion was 2.22 ng x g creat(-1) x kg BW(-1) x kg (p.e.)(-1). PMID- 11910468 TI - Cleanup of gloves contaminated with granular terbufos and tefluthrin. AB - Chemical-resistant gloves are used for protection from pesticides in farming operations. Cleanup of gloves after pesticide contamination was the focus of this research. Nitrile, neoprene, and barrier laminate glove specimens were exposed to 300 mg terbufos or tefluthrin granules for 3 or 30 min in petri dishes in a laboratory. Specimens were cleaned by flush with running water or LaunderOmeter washing with detergent. Following the cleanup treatments, specimens were dried and placed in test tubes with solvents to extract pesticide residue. Levels of contamination remaining were determined by gas chromatography. The residue remaining varied with exposure time, material type, cleanup method, and pesticide. Flush was more effective with the shorter exposure time. Tefluthrin was more effectively removed than terbufos. Barrier laminate was confirmed as a single-use material. Cleanup procedures reduced contamination in nitrile and neoprene, but findings show that these materials retained residue after cleanup. PMID- 11910469 TI - Smoking is a risk factor for recurrence of groin hernia. AB - Studies of connective tissue from patients with inguinal hernia have shown that smoking may be associated with hernia formation due to a defective connective tissue metabolism. Whether smoking is a risk factor for recurrence, too, was examined in this study. From December 1990 through December 1995, 649 patients underwent hernia repair as open sutured repair (Cooper ligament or abdominal ring repair) or as open mesh repair. Five hundred forty-four eligible patients were evaluated for recurrence 2 years postoperatively. Association between recurrence and 17 patient-, disease-, and intraoperative variables were analyzed by multiple logistic regression. The results showed that smoking was significantly and independently associated with recurrence compared to nonsmoking [odds ratio (OR = 2.22; 95% confidence interval (95% CI) = 1.19-4.15)]. Open sutured repair compared to open mesh repair was the most significant predictor for recurrence (OR = 7.23; 95% CI = 3.01-17.37). Surprisingly, local anesthesia was associated with a higher risk of recurrence compared to general anesthesia (OR = 2.44; 95% CI = 1.19-5.09). Potential confounders and other risk factors for hernia recurrence such as age, alcohol consumption, previous surgery, and anatomical characteristics of the hernia were adjusted for in the analysis. In conclusion, smoking is an important risk factor for recurrence of groin hernia, presumably due to an abnormal connective tissue metabolism in smokers. PMID- 11910470 TI - Recurrent inguinal hernia: disease of the collagen matrix? AB - The aim of this study was to investigate the collagen matrix in recurrent inguinal hernias. Total ribonucleic acid (RNA) was extracted from skin fibroblasts of three groups (control group I = healthy skin; control group II = plain skin scar; recurrent inguinal hernia group = skin of recurrent inguinal hernias; each n = 5). Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis were used to investigate the expression of procollagen type I/- III, MMP-1, and MMP-13 mRNAs. Both ratios of procollagen types I to III mRNAs and collagen types I to III were apparently decreased in the recurrent hernia group compared to those of both control groups (p < 0.01). Significant differences were caused by the increase of both procollagen type III mRNA and collagen type III protein synthesis. A concomitant increase of MMP-1 and MMP-13 mRNAs and proteins was also observed in the recurrent hernia group and showed significant differences compared to those of both control groups I and II, respectively (p < 0.01). In conclusion, the decreased ratio of collagen types I to III seems not only to be the result of a relative increase in the levels of type III procollagen mRNA but also may be the result of an increase of MMP-1 and MMP-13. The data of the present study strongly suggest recurrent inguinal hernias to be a disease of the collagen matrix and result in a clearer understanding of the underlying pathophysiology and may support specific therapeutic strategies in hernia surgery (e.g., surgical meshes). PMID- 11910471 TI - Experimental evaluation of a new layered prosthesis exhibiting a low tensile modulus of elasticity: long-term integration response within the rat abdominal wall. AB - The use of a new type of prosthesis, Bard Composix (BC), constructed of two layers of polypropylene mesh (PP) and one layer of expanded polytetrafluoroethylene (ePTFE), could provide a good solution for hernia repair when both minimal adhesions and maximum collagenous infiltration are necessary. We experimentally evaluated long-term stability of this composite. In 15 Sprague Dawley rats, a full thickness defect was created in the anterior abdominal wall and repaired with BC. Studies were performed over implantation intervals of 2, 4, and 6 months in strips obtained from the prosthesis-host tissue interfaces. Light microscopy, environmental scanning electron microscopy (ESEM), immunohistochemistry, and tensiometry were used. Overall findings provide evidence that PP and ePTFE association renders the alloy well suited for hernia repair, promoting a robust and durable alloplast-soft tissue union. At all points studied, the patch was well tolerated and meshes did not shrink, come loose, or migrate. Neovascularization continued 6 months after implantation. Ex vivo mechanical characterization demonstrated that the primary advantage of the new device stems from a low modulus of elasticity, a property that can be exploited to enhance mechanical load transfer from prosthetic materials to the relatively frail surrounding tissues. After implantation, adequate tensile strength and a low modulus of elasticity were detected in the restored zone, conferring great adaptability to the abdominal wall. In conclusion, the BC layered prosthesis proved suitable for implantation in abdominal wall defects, exhibiting favorable biocompatibility and integration with minimal side effects. PMID- 11910472 TI - Factors affecting management and outcome in blunt renal injury. AB - Blunt renal trauma patients often have multiple injuries. We retrospectively analyzed factors that could be predictive of outcome and the need for nephrectomy in this population. Blunt renal injury patients admitted to a Level I trauma center from January 1989 to July 1997 were identified and their charts reviewed. Multiple factors were examined. Using logistic regression analysis, factors predictive of mortality and the need for nephrectomy were determined. Of 11,847 trauma patients admitted, 95 (0.80%) suffered blunt renal injury. Mean age and ISS were 31.4 and 23.7, respectively. The number of deaths and nephrectomies was 11 and 10, respectively. Higher renal injury grade, as well as higher ISS values and 24-hour transfusion needs, directly correlated with the need for nephrectomy. Greater age, higher ISS, and higher 24-hour transfusion requirements lowered probability for survival. Patients with blunt renal injuries often sustain multiple injuries. The grade of renal injury, the overall injury severity of the patient, and the requirement of blood transfusion are the primary factors in determining the patient's need for nephrectomy and overall outcome. PMID- 11910473 TI - Connective tissue growth factor gene expression alters tumor progression in esophageal cancer. AB - The ability of cancer cells to initiate specific fibroblast reactions may subsequently determine tumor evolution. In the present study we examined the coordinated expression of transforming growth factor-beta-1 (TGF-beta1), its signaling receptors, and its downstream mediator-connective tissue growth factor (CTGF)--and their impact on tumor progression and fibrogenesis in esophageal carcinomas. Messenger ribonucleic acid (mRNA) expression of TGF-beta1, CTGF, TGF beta receptor subtype I ALK5 (TbetaR-IALK5), and TGF-beta receptor type II (TbetaR-II) was studied by Northern blot analysis in esophageal cancer and the normal esophagus. By means of immunohistochemistry and Western blot analysis, the respective proteins were localized in the tissue samples and the protein content was quantitated. Northern blot analysis revealed 3-fold and 4-fold increases (p < 0.05) in TGF-beta1 and CTGF mRNA levels, respectively, in esophageal cancer in comparison with normal controls, whereas TbetaR-I mRNA levels were significantly decreased and TbetaR-II mRNA levels were unchanged in the cancer samples. Immunostaining revealed results similar to those seen on the RNA level. TGF-beta1 and CTGF immunoreactivity were increased, TbetaR-II was unchanged, and TbetaR IALK5 immunoreactivity was decreased. CTGF immunoreactivity was mainly present in the stroma surrounding the cancer cells but was also present in the cancer cells. The degree of fibrosis was different in squamous and adenocarcinomas and was significantly related to CTGF mRNA expression levels. The presence of CTGF in squamous cell carcinomas was associated with longer survival, whereas in adenocarcinomas it influenced survival negatively. The findings indicate that TGF beta signaling is disturbed in esophageal cancer. CTGF, a downstream effector of TGF-beta action, differentially influences the composition of tumor microenvironment and distinct cell-matrix interactions in the two histological types of esophageal carcinoma, resulting in differences in tumor progression and patient survival. PMID- 11910474 TI - Re-study of gastric cancer: analysis of outcome. AB - Cancer of the stomach (CaS) is a dreaded disease. Fortunately, there is a decreasing incidence, except in the East. The authors did a re-study of CaS, a widely investigated but unresolved gastrointestinal malignancy. The clinicopathologic features were evaluated to identify and measure the prognostic factors that would help the surgeon decide optimal therapy. Among 383 admitted for CaS at the East Avenue Medical Center, Quezon City, Philippines between January 1987 and December 1996, 149 underwent radical resection with curative intent. (As historical control, the experience in 136 cases was reviewed during the immediately preceding 5-year period [1982-1986] when extended lymphadenectomy was not the standard policy.) For staging, the TNM system (tumor-node-metastasis) was used; to describe anatomy and surgery of stomach lymphatics, the "Japanese Rules," as modified, were adapted. Curative radical gastrectomy would include removal of the diseased stomach and regional lymphatics as defined by frozen section, including subtotal (or total) gastrectomy and "extended" D2 (with no. 12) node dissection. The clinicopathologic factors were statistically analyzed, using the accepted methods: Kaplan-Meier for survival, univariate analysis, and multivariate analysis for independent predictors. Of the 12 risk factors assessed by univariate analysis, the following were identified by multivariate analysis as independent prognosticators of survival: (1) wall penetration; (2) node invasion; (3) TNM stage; (4) resection margin; and (5) tumor size. After curative resection, the operative mortality was 5.3% and the complications, 19.4%. The 5 year survival was 60.4%, and recurrence, 15.4%. The results have shown that the pathology-related factors, (1) wall penetration; (2) node invasion; and (3) resection margin, are independent prognosticators of survival, remarkably affecting outcome. In conclusion, the study supports radical gastrectomy with extended D2 lymphadenectomy for CaS as safe and effective. Survival and recurrence are a function of pathology and adequate resection; operative mortality is defined by the patient's condition. PMID- 11910475 TI - One-stage surgical management of concomitant abdominal aortic aneurysm and gastric or colorectal cancer. AB - One-stage surgical management of concomitant abdominal aortic aneurysm (AAA) and gastric or colorectal cancer should provide certain benefits. We reviewed the records of 21 patients with both AAA and gastric or colorectal cancer who underwent one-stage surgical management. Four had distal gastrectomy, 2 had total gastrectomy, and 5 had abdominoperineal rectal resection transperitoneally; 3 had total gastrectomy transperitoneally and AAA repair extraperitoneally. Two underwent right hemicolectomy and thromboexclusion of the AAA. Two had creation of a temporary ileostomy and implantation of an interposition graft. Two underwent left hemicolectomy, creation of a temporary transversostomy, and implantation of an interposition graft. One had a Hartmann's procedure and implantation of a bifurcated prosthetic interposition graft for AAA. There were no operative deaths or serious postoperative complications. One patient had colorectal ischemia that resolved with conservative treatment. Eighteen of the 21 patients (85.7%) were alive 10 months to 14 years postoperatively. In conclusion, one-stage surgical treatment of concomitant AAA and gastric or colorectal cancer is well tolerated and can avoid the time, financial costs, and patient anxiety involved in a second operation. PMID- 11910476 TI - Small bowel intussusception in symptomatic pediatric patients: experiences with 19 surgically proven cases. AB - Nineteen cases of surgically proven symptomatic pediatric small bowel intussusceptions (SBI) were retrospectively reviewed. Clinical presentations included vomiting (89.5%), abdominal pain and/or irritable crying (89.5%), fever (52.6%), bloody stools (26.3%), palpable abdominal masses (15.8%), hematemesis (10.5%), jaundice (5.3%), and seizures (5.3%). The duration between symptom onset and hospitalization ranged between 20 and 336 hours (average 75.8 hours). Two patients with suspected appendicitis and small bowel obstruction were operated on promptly. Sonograms revealed target lesions (average diameter 2.9 cm) suggestive of intussusception in 13 out of 17 patients, with 10 lesions located in the paraumbilical or left abdominal regions. Barium enemas in 12 of these 13 patients demonstrated no colonic lesions. Diagnosis and surgery were delayed in 16 patients (average delay = 32 hours). The remaining 1 patient with positive sonographic findings underwent early surgery after computed tomographic (CT) confirmation of SBI. Surgery revealed ileoileal intussusceptions in 11 patients, jejunojejunal in 4, jejunoileal in 3, and duodenojejunal in 1. Eight patients had lead points. Bowel complications (ischemia, necrosis, or perforation) occurred in 8 patients. The duration between symptom onset and surgery in patients with bowel complications was significantly longer than for patients without complications (p = 0.0026). In conclusion, delayed diagnosis and surgical treatment in symptomatic pediatric patients with SBI were common, leading to a high rate (42%) of bowel complications. Sonographic demonstration of a 2-3 cm target lesion, especially if paraumbilical or left abdominal, is suggestive of SBI and may obviate the need for a barium enema; however, CT is helpful for confirming SBI. In symptomatic SBI, once diagnosed, early surgical referral is strongly recommended. PMID- 11910477 TI - Surgical indications and strategies for necrotizing enterocolitis in low income countries. AB - In high income countries, mortality of neonates due to necrotizing enterocolitis (NEC) is decreasing due to advances in early detection and sepsis management. Surgical strategies, by and large, are becoming less relevant as the sophistication of intensive care facilities increases in those countries. Management of septic neonates with NEC in low income countries, however, due to inherent resource constraints cannot achieve the same standards as those reported from high income countries. Surgeons therefore play an important role because they can offer different surgical strategies in line with the deficiency of their environment. Over a 4-year period, 172 neonates who had surgery for NEC in a single institution were prospectively followed up. Standard surgical practices of the high income countries were compared to those adopted by the authors. The end point was early mortality, taking into account the anatomical zones of the involved intestine. Survival was increased from 21% to 84% by early surgery and by extending the length of colon resected in cases of ileocolic NEC. In this approach, a short resuscitative period is followed by intestinal resection including most of the diseased colon as well as the perforated/dead bowel. A more modest improvement in survival was noted with a planned re-look laparotomy in neonates with generalized NEC. PMID- 11910478 TI - Effect of bile acid replacement on endotoxin-induced tumor necrosis factor-alpha production in obstructive jaundice. AB - There is a high incidence of perioperative morbidity and mortality in patients with obstructive jaundice due to sepsis. Tumor necrosis factor-a (TNF-alpha) is considered a crucial mediator in inducing and processing the inflammatory cascade. We hypothesize that obstructive jaundice leads to an increased endotoxin induced TNF-alpha production and that intestinal bile acid replacement can prevent this phenomenon. Sprague-Dawley rats were randomized to three groups of 12 animals each. Group 1 underwent common bile duct ligation (CBDL) with oral intestinal bile acid (deoxycholic acid 5 mg/100 g body weight/3 times daily) replacement (CBDL + bile acid); group 2 underwent common bile duct ligation with the same amount of normal saline replacement orally (CBDL + saline); and group 3 underwent a sham operation (sham control). After 2 days, endotoxin was given to the animals, and after 90 minutes, tissues (liver and lung) and blood were collected for checking the TNF-alpha levels and biochemical analyses. Comparisons among these three groups were performed and recorded. While serum and tissue (liver and lung) TNF-alpha levels of group 2 (CBDL + saline) were significantly increased after endotoxin challenge, these elevations were reduced to control levels (sham control) following oral replacement of intestinal bile acid (CBDL + bile acid). Obstructive jaundice leads to an increased endotoxin-induced TNF alpha production and intestinal bile acid replacement can inhibit this phenomenon. PMID- 11910479 TI - Significance of serum type IV collagen level of hepatectomized patients with chronic liver damage. AB - Type IV collagen, one of the serum markers for hepatic fibrosis, was measured perioperatively in patients with and without chronic liver damage to investigate whether this parameter changes in response to acute stress to the liver and can predict the surgical risk of hepatic resection. The serum type IV collagen level was significantly elevated in patients with liver cirrhosis. There were significant correlations between serum type IV collagen levels and the indocyanine green clearance test and cholinesterase activity, although the correlation coefficients were not high. The size of the resected hepatic mass was not the primary factor to influence the postoperative serum type IV collagen level. In patients with liver cirrhosis, the postoperative serum type IV collagen level increased significantly compared to that in patients with normal liver or chronic hepatitis. Postoperative liver failure occurred in 0%, 11.6%, and 44.4% of patients with preoperative serum type IV collagen levels of <150, < or = 150 to 300, and > or = 300 ng/ml, respectively. In those with postoperative liver failure, the serum type IV collagen levels were significantly higher both pre- and postoperatively compared to those in patients with uneventful courses. Several preoperative liver function tests indicated that type IV collagen is an independent risk factor for postoperative liver failure. Thus perioperative measurement of the serum type IV collagen levels seemed to be useful for predicting the risk of hepatic resection in patients with chronic liver damage. PMID- 11910480 TI - Relation of biliary bile acid output to hepatic adenosine triphosphate level and biliary indocyanine green excretion in humans. AB - We have previously demonstrated that there are two subgroups of patients with different types of biliary bile acid output after relief of obstructive jaundice by percutaneous transhepatic biliary drainage (PTBD). The reason for two groups is not clear but is possibly the difference in hepatic reserve function. The aim of this study was to examine the relation of biliary bile acid output to the hepatic ATP level and biliary excretion rate of indocyanine green (ICG) in humans. Patients whose bile could be collected through a PTBD tube participated in this study. The mean serum total bilirubin concentration was 12.7 mg/dl at the time of PTBD, decreasing to 1.1 mg/dl before surgery. These patients underwent curative resection for cancer of the bile duct, duodenal papilla, or pancreatic head after the relief of hyperbilirubinemia. Bile was collected at 1-hour intervals for 5 hours after intravenous administration of ICG (0.5 mg/kg) within a few days before surgery, and a small liver tissue sample was obtained immediately after laparotomy without using ischemic procedures. The concentrations of total bile acid and ICG in bile, the bile flow rate, and the bile acid output and ICG excretion rate in bile over 5 hours were determined. ATP concentrations in liver tissue were determined by high performance liquid chromatography. Results of hepatic ATP levels were correlated with the bile acid output and ICG excretion rate into bile. Both the biliary bile acid output (micromoles per 5 hours) and ICG excretion rate (percent of injected dose of ICG) over 5 hours were significantly correlated with the hepatic ATP level (p = 0.0190 and p = 0.0084, respectively). Neither the bile flow rate nor the serum liver function tests were related to the hepatic ATP level. Significant correlation was found between the bile acid output and the ICG excretion rate (p = 0.0127). Biliary bile acid output reflects the hepatic ATP level. Determination of the biliary bile acid output and ICG excretion may provide useful parameters for evaluating the hepatic energy status, which is essential for organ viability. PMID- 11910481 TI - Surgical strategy for cystic diseases of the liver in a western hepatobiliary center. AB - The aim of this study was to define the indications and evaluate the results of various management options in patients with cystic liver disease. Between 1992 and 1999 we managed 60 consecutive patients with cystic liver disease. Diagnoses included a simple cyst (solitary 12, multiple 10), adult polycystic liver disease (APLD 17), Caroli's disease (8), hydatid cysts (4), and neoplastic cysts (9). Half of the patients with simple cysts had mild or no symptoms and required no treatment. Percutaneous drainage in eight patients (simple cyst 4, APLD 4) was followed by symptomatic recurrence in three. Laparoscopic deroofing in three patients (multiple simple cysts 2, APLD 1) was followed by symptomatic enlargement of the remaining cysts that required further intervention (laparoscopic deroofing 2, transplantation 1). Laparoscopic hepatectomy was successful in three patients with solitary simple cysts. Of 18 patients who underwent open hepatic resection (neoplastic 8, Caroli's 4, simple cysts 3, hydatid cysts 2, APLD 1), 2 patients with Caroli's disease required liver transplantation for disease progression. Nine patients (Caroli's 5, APLD 4) underwent liver transplantation, and three had a concomitant renal transplant. Seven patients developed complications, and three died (5%). Cholangiocarcinoma developed in three patients with bilateral Caroli's disease, and all died. Radiologic treatment has a limited role in the management of patients with simple cysts or APLD. Laparoscopic deroofing of simple cysts may have to be repeated, whereas resection minimizes cyst recurrence. Unilobar Caroli's disease may be resected, whereas bilateral disease requires early liver transplantation owing to the high risk of malignancy. Transplantation is a reserved option in patients with extensive APLD. PMID- 11910482 TI - Incidence of deep venous thrombosis in patients undergoing obesity surgery. AB - The aim of this study was to investigate prospectively the incidence of deep venous thrombosis (DVT) after surgery for morbid obesity. The series comprised 116 consecutive patients undergoing Roux-en-Y gastric bypass. The median age and body mass index were 35 years (range 19-59 years) and 42 kg/m2 (range 32-68 kg/m2), respectively. The patients were examined with duplex ultrasonography pre- and postoperatively. No patient had any symptoms or signs of DVT postoperatively, and ultrasonography showed no signs of thrombosis in iliac, femoral, and popliteal veins in any of the patients. Two patients (1.7%) had a thrombus in the peroneal vein of one leg. Repeated ultrasonographic investigation after 1 week showed complete resolution of both. One patient with a previously unknown activated protein C resistance had an angiographically confirmed minor pulmonary embolus. The incidence of venous thromboembolism after obesity surgery seems to be low, and obesity as a risk factor for thromboembolic disease might have been overestimated in the past. PMID- 11910484 TI - Pancreatic function after acute biliary pancreatitis: does it change? AB - According to the Cambridge and Marseilles symposia, morphologic and functional recovery from acute biliary pancreatitis (ABP) occurs if the initial cause and complications are eradicated. Nevertheless, in recent years there has been controversy over this topic, and varying results have been reported. These differing results may be due to different diagnostic methods, number of patients studied with regard to etiologic factors, severity of the disease, and differences in the tests used. A total of 63 ABP patients [17 male (27%), 46 female (73%); 45 mild, 18 severe] were prospectively studied. All patients underwent cholecystectomy. No patient in this series underwent necrosectomy or pancreatectomy. During the acute phase the severity of the disease was assessed following the Atlanta criteria, and the occurrence and rate of necrosis were determined by dynamic computed tomography. Exocrine and endocrine pancreatic functions were assessed at 1 month, 6 months, and 1 year after the ABP episode by means of various pancreatic function tests. We evaluated the occurrence of pancreatic failure following ABP and if this deficiency was associated with the severity of the episode. According to the classic symposia criteria, the study showed no deficit in exocrine or endocrine pancreatic function. No statistically significant differences were found when the pancreatic function and severity of the process were assessed. PMID- 11910483 TI - Surgical treatment for severe acute pancreatitis: extent and surgical control of necrosis determine outcome. AB - In patients operated on for severe acute pancreatitis (SAP), the factors determining outcome remain unclear. From 1986 to 1998 a total of 340 patients with a diagnosis of SAP and in need of operative treatment were admitted to the intensive care unit (ICU) of a university hospital and a secondary care hospital. The mean APACHE II score on the day of admission was 16.1 (range 8-35). All patients required operative therapy. Among the 340 patients, 270 (79.4%) had to be reoperated: 196 patients (72.6%) underwent operative revisions on demand, and 74 (27.4%) patients had preplanned reoperation. The overall mortality was 39.1% (133 patients). Septic organ failure in 126 patients (37.1%) and myocardial infarction or pulmonary embolism in 7 patients (2%) were the causes of death. The patient's age (p < 0.0002), APACHE II scores at admission (p < 0.0001), presence or development of (single or multiple) organ failure (p < 0.002), infection (p < 0.02) and extent (p < 0.04) of pancreatic necrosis, and surgical control of local necrosis (p < 0.0001) significantly determined survival. SAP that requires surgical treatment is associated with high in-hospital mortality. Surgical control of local necrosis is the precondition for survival. Advanced age of the patient, high APACHE II score at admission, development of organ failure, and the extent and infection of pancreatic necrosis influence the outcome. PMID- 11910485 TI - Intraoperative transfusion: is it a real prognostic factor of periampullary cancer following pancreatoduodenectomy? AB - The purpose of this study was to clarify the prognostic significance of transfusion following pancreatoduodenectomy for periampullary cancers. We analyzed 357 periampullary cancers from 1985 to 1997 (ampullary cancer 130 cases, distal bile duct cancer 141 cases, pancreatic head cancer 86 cases). A total of 215 (60%) of the 357 patients have received intraoperative transfusion. The 5 year survival rate of 130 ampullary cancer patients was 59%; altogether, 76 patients (58%) underwent intraoperative transfusion. The 5-year survival rate of patients without intraoperative transfusion was 79%, whereas that of patients with a transfusion was 47% (p = 0.029). Following multivariate analysis, intraoperative transfusion was found to be an independent poor prognostic factor for those with ampullary cancer (relative risk 2.174). Among those with common bile duct cancer, the overall 5-year survival rate was 33%, and the 5-year survival rates for patients with (n = 87) or without (n = 54) transfusion were 25% and 38%, respectively, which did not reach statistical significance (p = 0.0717). For those with pancreatic head cancer, the overall 5-year survival rate was 16%, and there was no survival difference between transfused (n = 52) and untransfused (n = 34) patients. In the present study the reason was not clear, although intraoperative transfusion was an independent significant prognostic factor for ampullary cancer. Careful dissection to minimize intraoperative bleeding is mandatory during pancreatoduodenectomy for ampullary cancer. PMID- 11910486 TI - Contractility in vitro and mitochondrial response in small and large rabbit bowel after anastomosis. AB - Leakage of a large bowel anastomosis remains the most serious postoperative complication in gastrointestinal surgery. In a recent experimental study we found that surgically induced hypoxia resulted in more derangement of a variety of biochemical markers in the large bowel (LB) than in the small bowel (SB). We explored the question of whether spontaneous and agonist-induced contractility of SB and LB muscle strips was influenced by surgical procedures and how contractility was related to energetic oxidative metabolism capacity in smooth muscle mitochondria. Sixty male New Zealand rabbits were operated on under general anesthesia. Segments of ileum and colon were resected from each rabbit, and an end-to-end anastomosis was constructed. A representative subset of segments from SB (n = 14) and LB (n = 14) at time 0 was used as controls. Tracts containing an anastomosis were resected at days 2, 7, and 14 after operation. At each time point, 20 segments adjacent to the anastomosis of both SB and LB were used for tensiometric and biochemical studies. Tensiometric studies demonstrated modifications in the smooth muscle function at both the acute and chronic stages with intestinal inflammation that may contribute to surgical stress-associated abnormal motility. Biochemical data showed that the respiratory capacity of the resected LB was more impaired than that of the SB. In both SB and LB, changes in respiratory activity preceded tensiometric changes. Thus abnormalities of contractility after surgical stress are more evident in LB than SB in segments adjacent to the anastomoses. This could be the consequence of abnormal biochemical changes, as adenosine triphosphate (ATP) is required for membrane potential maintenance, calcium homeostasis, and actin-myosin interactions. PMID- 11910487 TI - Factors associated with clinically significant anastomotic leakage after large bowel resection: multivariate analysis of 707 patients. AB - The aim of this study was to determine by univariate and multivariate analyses the factors associated with clinically significant anastomotic leakage (AL) after large bowel resection. From 1990 to 1997 a series of 707 patients underwent colonic or rectal resection (without a stoma). Patients were divided into two groups: those with clinical anastomotic leakage (group 1) and those without it (group 2). AL occurred in 43 of 707 patients (6%). The overall mortality was 2.2% and was significantly higher in patients with AL than in those without: 5 of 43 (12%) versus 11 of 664 (1.6%), p < 0.001. Univariate analysis showed 15 variables associated with the risk of AL: previous abdominal or pelvic irradiation (p = 0.02), American Society of Anesthesiologists (ASA) score > 2 (p = 0.04), leukocytosis (p = 0.02), renal failure (p = 0.03), steroid treatment (p = 0.01), duration of operation (p = 0.001), intraoperative septic conditions (p = 0.006), total colectomy (p = 0.009), transverse colectomy (p = 0.02), difficulties encountered during anastomosis (p = 0.001), ileorectal anastomosis (p = 0.02), colocolic anastomosis (p = 0.01), abdominal drainage (p = 0.05), and blood transfusion intraoperatively (p = 0.006) and postoperatively (p = 0.001). Multivariate analysis showed that only preoperative leukocytosis (p = 0.04), intraoperative septic conditions (p = 0.001), difficulties encountered during anastomosis (p = 0.007), colocolic anastomosis (p = 0.004), and postoperative blood transfusion (p = 0.0007) were independent factors associated with AL. The risk of AL increased from a range of 12% to 30% if one risk factor was present, to 38% with two factors, to 50% with three factors. After colorectal resection and intraperitoneal anastomosis, a temporary protective stoma is proposed in selected patients with high risk factors for AL, as observed in our study. PMID- 11910488 TI - Small nonpolypoid colorectal carcinoma. AB - The objective of this study was to characterize and assess the presence and frequency of small nonpolypoid colorectal adenocarcinomas among patients with colorectal cancer referred for surgery. The medical, endoscopic, and surgical reports and the histopathologic slides of all patients operated on for colorectal cancer were retrospectively reviewed. Small nonpolypoid colorectal cancer (SNPCC) was defined as a malignant, nonpolypoid lesion smaller than 15 mm. SNPCC was classified according to the Japanese macroscopic classification of colorectal carcinoma. The frequency of SNPCC among patients referred for operation was 1.8%. Most of these patients were asymptomatic and were diagnosed by the same endoscopist using a high-resolution video-endoscope without the assistance of enhancement techniques. These lesions had a mean size of 10.8 mm, were mainly of the flat or flat elevated type, and were located in the distal colon. Among patients with colorectal cancer referred for surgery, 1.8% had SNPCC. These lesions can be detected using high-resolution video-endoscopy equipment without the need for enhancement techniques, as reported in Japanese series. Increased awareness of the existence of such SNPCC lesions may help the average endoscopist detect such lesions. As SNPCC represents colorectal cancer, all the cases in our series were treated by typical oncologic surgical resection. PMID- 11910489 TI - Case series of acute abdominal surgery in rural Sierra Leone. AB - In many poor countries of the world the need for surgical treatment of acute abdominal emergencies is largely unmet. In some cases this service is provided by physicians with little postgraduate surgical training, and there is a paucity of published data on the outcomes of this service. This series of sequential cases of acute abdominal surgical emergencies from a hospital in rural Sierra Leone illustrates the causes, outcomes, and challenges in this setting. All patients with an acute abdomen from September 1992 until September 1994 who required surgery were identified by review of theater records, ward books, and patients' notes. Altogether, 173 cases were identified. Operative diagnoses included ectopic pregnancy (n = 43), strangulated hernia (n = 45) 15 of which required bowel resection, appendicitis (n = 15), normal appendix (n = 4), uterine rupture (n = 9), perforated ulcer (n = 8), tubal or pelvic abscess (n = 7), volvulus (n = 6), and others. Ninety percent survived to discharge after a median postoperative stay of 9.2 days (range 7-127 days). Of the 18 deaths, 83% occurred during the first 3 days. Factors associated with poor outcome were ileal perforation due to typhoid fever and resection of bowel after a strangulated hernia. These results show that acute abdominal surgery can be done at the district level in poor countries using limited facilities by staff without extensive surgical training. The outcomes are comparable to those from larger centers. PMID- 11910490 TI - F-like plasmid sequences in enteric bacteria of diverse origin, with implication of horizontal transfer and plasmid host range. AB - Seventy-eight bacterial isolates from human, animal, and plant hosts, representing eight species of the family Enterobacteriaceae, were screened for F like plasmid sequences. Of the examined human Escherichia coli strains, 28% harbored one or two of the three F-like, RepFI replication regions, while 35% of the examined animal and all phytopathogenic strains harbored RepFIA-specific sequences. Comparative analysis of Salmonella, Shigella, Erwinia, and E. coli plasmid RepFI sequences showed 100% or very high homology, indicating frequent and recent interspecies gene transfer. The high incidence of RepFIA sequences in enteric bacterial species, including Klebsiella and Erwinia, showed that F-like plasmids are successful in avoiding natural barriers to establishment of horizontally transferred DNA and that in the natural environment conjugal transfer is efficient in diverse ecological niches. PMID- 11910492 TI - Three-piece-ligation PCR and application in disruption of chlorophyll synthesis genes in Synechocystis sp. PCC 6803. AB - Linear DNA, consisting of a drug-resistance marker and long flanking sequences, was synthesized by one-step polymerase chain reaction after a three-piece ligating reaction. Chlorophyll synthesis genes, chlH and chlL in Synechocystis sp. PCC 6803, were replaced by a kanamycin-resistance marker through double recombinations with flanking homology regions. Under LAHG conditions, the chlL but not chlH mutant stopped chlorophyll synthesis, while both synthesized chlorophyll in the light. PMID- 11910491 TI - Mapping the fibrinogen-binding domain of serum opacity factor of group a streptococci. AB - Serum opacity factor (SOF) is a large, extracellular, and cell-bound protein of group A streptococci that has two known functions, opacification of serum and binding of fibronectin. Herein, we describe a new function of SOF, the binding of fibrinogen. Utilizing purified, truncated recombinant SOF proteins, the fibrinogen-binding domain was localized to a region in the C-terminus of SOF encompassing amino acid residues 844-1047. Western-blot analysis revealed that SOF bound primarily to the beta subunit of fibrinogen. A SOF-negative mutant bound 50% less fibrinogen than did its wild-type parent. Furthermore, fibrinogen blocked the binding of SOF to fibronectin. These data suggest that fibrinogen and fibronectin bind to the same domain within SOF. It remains to be determined whether the binding of fibrinogen to SOF contributes to the virulence of group A streptococci. PMID- 11910493 TI - Characterization of an alanine racemase gene from Lactobacillus reuteri. AB - An alanine racemase gene from Lb. reuteri was cloned by using degenerate oligonucleotides corresponding to conserved regions derived from several bacterial alanine racemases. The protein is 375alphaalpha in length and shows 63.6% homology to the Lb. plantarum alanine racemase. Unlike the single alanine racemase activity found in Lb. plantarum, deletion of the Lb. reuteri alanine racemase reveals a second activity, which is inhibited by beta-chloro- D-alanine. PMID- 11910494 TI - Recycle use of Sphingomonas sp. CDH-7 cells for continuous degradation of carbazole in the presence of MgCl2. AB - Carbazole (CA) is a heterocyclic nitrogen compound contained in the crude petroleum oil and recalcitrant to removal through the refinery processes. For development of the efficient CA-degradation bioprocess, conditions for the recycle use of Sphingomonas sp. CDH-7 resting cells were examined. When the resting cells (O.D.(660) 3.3) were shaken in 50 m M K2HPO4-KH2PO4 buffer (pH 7.0) containing CA 1000 mg/L, CA 880 mg/L was degraded within 3 h, but thereafter the activity decreased markedly. However, the activity was found to be restored to the initial level after the shaking treatment for 3 h in CA-free medium solution or in the buffer containing 20 m M MgCl2. Although the CA-degradation activity of CDH-7 resting cells was lost after 3 h of shaking in the buffer containing 100 m M EDTA, it was restored through the shaking treatment for 3 h in the buffer containing 20 m M MgCl2. When CA was periodically added eight times at a concentration of 100 mg/L (0.599 m M) to the reaction mixture containing the resting cells, CA 778 mg/L (4.66 m M) was continuously degraded within 35 h by the recycle use of resting cells, with the restoration treatment after each CA degradation reaction by the resting cells. PMID- 11910495 TI - Urate oxidase from the rust Puccinia recondita is a heterotetramer with two different-sized monomers. AB - Uricase (urate: oxygen oxidoreductase; EC 1.7.3.3) from the rust Puccinia recondita was purified to electrophoretic homogeneity. Preparations with a specific activity of 8.4 U/mg were used for characterization of the enzyme, which showed a strong similarity to other plant and fungal urate oxidases. The enzyme had a pH optimum of 9.0, a K(m) of 35 microM for urate, and it was inhibited only by oxonate and xanthine. A molecular mass of 152 kDa was estimated for the native protein. SDS-PAGE analysis revealed a striking difference to most urate oxidases, since two different-sized subunits were detected. These results suggest that P. recondita uricase is a tetramer with two types of subunits. PMID- 11910497 TI - Characterization of the Porphyromonas gingivalis FtsZ containing a novel GTPase activity. AB - The FtsZ protein is a GTPase that is essential for cell division. We have cloned, sequenced, and expressed the FtsZ (PgFtsZ) gene from the Porphyromonas gingivalis, an oral, anaerobic, rod-shaped bacterium implicated in progressive periodontal disease. The PgFtsZ gene consisted of 1374 bp and coded for an acidic protein with a calculated molecular mass of 50,253 Da. The deduced amino acid sequence exhibited a significant homology with E. coli FtsZ (54% identical residues). Like other prokaryotic FtsZs, PgFtsZ possessed the clear motifs for GTP binding (GGGTGTG) and hydrolysis (NLDFADV). When PgFtsZ was overexpressed in E. coli, cell division was inhibited. Recombinant PgFtsZ was purified to homogeneity and characterized. The purified PgFtsZ exhibited GTPase activity even in the absence of Mg2+, and completely retained its activity with EDTA. Furthermore, Na+ and K+ ions inhibited its GTPase activity in a dose-dependent manner. These results suggest that PgFtsZ contains an atypical GTPase activity that has not been previously described. PMID- 11910496 TI - Repressed respiration of oral streptococci grown in biofilms. AB - The respiratory activities of oral streptococci grown in biofilms were found to be markedly repressed compared with those of cells from aerobic culture, or for Streptococcus mutans GS-5, even for those grown in static culture. Respiration rates generally reflected levels of NADH oxidase activities in cell extracts. Superoxide dismutase levels were somewhat reduced in biofilm cells. However, sensitivities to oxidative damage caused by H2O2, t-butylhydroperoxide, or 8 hydroxyquinoline were not greatly different for cells from suspension cultures and those from either intact or dispersed biofilms. The capacities of S. sanguis and S. gordonii to produce H2O2 also were markedly repressed by biofilm growth, and presumably this repression would affect the ecology of dental plaque by reducing oxidative stresses under crowded conditions. PMID- 11910498 TI - Construction of double-copy glucose isomerase gene engineering strain of Streptomyces diastaticus by homologous recombination. AB - The plasmid pUT for homologous recombination was constructed by the insertion of the 1.1-kb thiostrepton resistance ( tsr(R)) gene into the E. coli plasmid pUB1 GI1. Plasmid pUTK was produced through ligating the cleaved plasmid pUT by KpnI. After pUT and pUTK were introduced into Streptomyces diastaticus No.7 strain M1033 (SM33) by protoplast transformation, a series of tsr(R) transformants were obtained, further based on enzyme assays. These results for polymerase chain reaction (PCR), DNA sequencing, restriction enzyme digestion, and recovery of cloned fragments from the transformant chromosome demonstrated the plasmid pUT and pUTK had integrated into the SM33 chromosome in three different patterns of single cross-over by homologous recombination. This directly results in double copy GI gene in the transformant chromosome, of which one is wild-type GI gene, the other mutant GI ( GIG138P, GI1) gene. Among the strains of the three kinds of recombinant patterns, one transformant was chosen and named K1, T2, and T3, respectively. The further identification of the three recombinant strains by PCR, DNA sequencing, restriction enzyme digestion, and Southern hybridization also proved there is a double-copy GI gene within their chromosome. Enzyme activity assay and thermostability analysis indicated that all three engineering strains expressed not only wild-type enzyme but also mutant GI. PMID- 11910499 TI - Characterization of a new Bacillus thuringiensis isolate highly active against Cochylis hospes. AB - A new bacterial isolate, 00-50-5, from sunflower head extracts was identified as Bacillus thuringiensis (Bt) according to its morphology. Bt isolate 00-50-5 was highly active against the banded sunflower moth (BSM), Cochylis hospes Walsingham. A sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) 4-15% gradient gel of whole strain protein of 00-50-5 revealed six proteins with molecular masses (Mr) of 133, 80, 60, 27, 15, and 14 kDa. SDS-PAGE of pH 4.2 precipitated proteins (PP) or activated proteins formed by adding the BSM larval gut protease at 1:50 (wt/wt, protease/PP) showed five bands, including two major proteins of Mr 60 kDa and 27 kDa, and three small peptides of Mr 15, 13, and 7 kDa. The BSM larval gut protease was able to completely digest the proteins when present at a high ratio (10:1, wt/wt, protease/PP). The 60- and 27-kDa proteins could be digested by subtilisin Carlsberg at ratios of 1:50 or 1:1 (wt/wt, protease/PP), but neither BSM larval gut protease nor trypsin was effective at the same ratios. Three small peptides of Mr 15, 13, and 7 kDa were digested by the gut protease at a ratio of 1:1 (wt/wt). The N-terminal sequence of 1-31 amino acid residues for the 27-kDa protein showed 96.7% homology to a 31-amino acid fragment from camelysin, a protease from B. cereus, indicating that the 27-kDa protein may be a camelysin and a novel active protein against BSM. PMID- 11910500 TI - Optimization of alkaline protease production from Bacillus sp. by response surface methodology. AB - High yields (1939 U/ml) of an alkaline protease were obtained in batch fermentation of a Bacillus sp. using a response surface methodology. The interaction of four variables, viz., starch, peptone, incubation time, and inoculum density, suggested inoculum density to be an insignificant variable. However, incubation time had a profound effect on protease yields at all the concentrations of carbon and nitrogen used. The response surface raised and flattened with increase in time of incubation, and maximum protease production up to 1939 U/ml was obtained after 96 h of incubation. The model equation obtained was validated experimentally at maximum starch (15 mg/ml) and peptone (7.5 mg/ml) concentration with increased incubation time up to 144 h in the presence of minimum inoculum density (1%). An overall 2.6-fold increase in protease production was obtained as compared with mean observed response (750 U/ml) at zero level of all variables. PMID- 11910501 TI - HtpG plays a role in cold acclimation in cyanobacteria. AB - Abstracts. The heat shock protein HtpG is homologous to members of the Hsp90 protein family of eukaryotes and is essential for basal and acquired thermotolerances in cyanobacteria. In this study we have examined the role of HtpG in the cyanobacterium, Synechococcus sp. PCC 7942, in the acclimation to low temperatures. The inactivation of the htpG gene resulted in severe inhibition of cell growth and of the photosynthetic activity when the htpG mutant was shifted to 16 degrees C from 30 degrees C. Wild-type cells were able to resume growth without a lag period when shifted to 30 degrees C after 5 days at 16 degrees C, while the mutant displayed a detectable lag. The HtpG protein was induced in the wild-type cells at 16 degrees C. Electrophoresis in the absence of sodium dodecyl sulfate (SDS) showed that a novel, high-molecular-weight complex containing GroEL and DnaK accumulated at 16 degrees C, but the accumulation was strongly inhibited in the htpG mutant. Our results demonstrate that the HtpG protein contributes significantly to the ability of cyanobacteria to acclimate to low temperatures. PMID- 11910502 TI - Prevalence of a novel capsule-associated lipoprotein among pasteurellaceae pathogenic in animals. AB - Capsular serotype A strains of Pasteurella multocida of avian origin express a 40 kDa lipoprotein (Plp-40) thought to attach the extracellular polysaccharide to the cell surface. The objective of the present study was to assess the prevalence of Plp-40 in P. multocida strains of disparate serotypes and host origins, as well as other pathogenic members of the family Pasteurellaceae. Exponential-phase reference and clinical isolates were radiolabeled with [3H]-palmitate, lysed to obtain whole-cell protein fractions, and analyzed using SDS-PAGE and fluorography to assess lipoprotein content. The ability to produce Plp-40 was found to be conserved among certain P. multocida reference and clinical strains of different host origins including avian, human, porcine, bovine, feline, canine, ovine, and cervine, but not rabbit. Production of a 40-kDa lipoprotein was exhibited by all clinical isolates of Pasteurella aerogenes, Pasteurella pneumotropica, Actinobacillus suis, Actinobacillus suis-like organism, and Actinobacillus pleuropneumoniae examined, but not Pasteurella (Mannheimia) haemolytica, Actinobacillus lignieresii, or Haemophilus spp. These data suggest that, while not all Pasteurellaceae are able to produce a 40-kDa lipoprotein under the present experimental conditions, expression is somewhat conserved among diverse isolates of disparate host origins. PMID- 11910503 TI - Levels of thermostable direct hemolysin produced by Vibrio parahaemolyticus O3:K6 and other serovars grown anaerobically with the presence of a bile acid. AB - Twenty-three V. parahaemolyticus strains, including 12 pandemic O3:K6 strains, were examined for their growth and production of thermostable direct hemolysin (TDH) under an anaerobic culture condition with or without presence of a bile acid, taurocholic acid (TCA). Both bacterial growth and TDH production were markedly enhanced by TCA for a majority of the strains, but the scale of the TDH production was disproportionately greater than that of the corresponding growth for 14 strains. Such enhancement was, however, not specific to the pandemic strains. PMID- 11910505 TI - What's in fibrin? PMID- 11910506 TI - Molecular mechanisms of human embryogenesis: developmental pathogenesis of renal tract malformations. AB - The focus of this review is the normal and abnormal development of the kidney and lower urinary tract; for convenience, we will refer to the whole system as the renal tract. The content represents a convergence among the clinical disciplines of histopathology, nephrology, and urology as well the basic sciences of developmental biology and molecular genetics. The story has considerable clinical relevance since diverse renal tract malformations are increasingly detected on fetal ultrasound screening and constitute major causes of chronic renal failure necessitating dialysis and kidney transplantation in children. Evidence is emerging that at least some of these disorders have a defined genetic basis; in others, an abnormal embryonic, or even maternal, environment may contribute to the pathogenesis. This field of study is frequently updated, with new discoveries being made almost every week. Hence this review can not be exhaustive or definitive, but instead highlights some specific areas of interest. PMID- 11910507 TI - Macrophage activation and hemophagocytic syndrome in langerhans cell histiocytosis: report of 30 cases. AB - Macrophage activation and secondary hemophagocytic syndrome are rarely reported in association with Langerhans cell histiocytosis (LCH). The authors reviewed their pathology files for cases of LCH in which evidence of macrophage activation coexisted and report 30 such cases indicating that the association is not that rare and may even be underdiagnosed unless specifically sought. Available clinical data were collected and correlated with pathological findings. Of the 30 cases of LCH with varying degrees of macrophage activation, 29 had multisystem disease. The cases were graded from I to V on the basis of evidence for, and severity of, macrophage activation; cases in category I had evidence of fully developed hemophagocytic syndrome whereas those in category V had limited evidence of macrophage activation. There were seven cases with fully developed hemophagocytic syndrome (category I) and an additional five with hemophagocytosis and some but not all of the features of hemophagocytic syndrome (category II). Most of these 12 cases were young children with high-risk LCH and poor prognosis; 4 are known to have died. Coexisting hemophagocytic syndrome in these cases of LCH may have contributed to their poor prognosis. The association of LCH with macrophage activation, though more than coincidental, is of unknown pathogenesis, but the role of T lymphocytes and cytokines is prominent in both disorders and is presumed to link the two. PMID- 11910508 TI - Lung pathology in premature infants with Ureaplasma urealyticum infection. AB - Respiratory tract colonization with Ureaplasma urealyticum in preterm infants has been associated with a higher incidence of pneumonia, severe respiratory failure, bronchopulmonary dysplasia (BPD), and death. In this report, we characterize the lung pathology and expression of tumor necrosis factor-alpha (TNF-alpha) associated with U. urealyticum pneumonia in very low-birth weight infants (VLBW; < or =1500 g). Lung pathology of archived autopsy specimens was retrospectively reviewed in three groups of VLBW infants: 5 gestational controls who died from nonpulmonary causes, 13 infants with pneumonia who were culture and/or PCR negative for U. urealyticum, and 5 infants with pulmonary disease and positive for U. urealyticum by tracheal aspirate and/or lung tissue culture or polymerase chain reaction (PCR). Presence and extent of alveolar macrophages and neutrophils, as well as interstitial lymphocytic infiltration and fibrosis were evaluated. PCR was performed on formalin-fixed, paraffin-embedded lung sections. Additional sections were immunostained for TNF-alpha. The peripheral total white blood cell counts and absolute neutrophil counts were three-fold higher in infants with U. urealyticum pneumonia than cell counts in infants infected with other organisms. There was a trend toward a predominance of neutrophils in alveoli of non- Ureaplasma pneumonia infants, but a trend toward a predominance of alveolar macrophages in U. urealyticum-infected infants. The most striking finding was the presence of increased interstitial fibrosis in all Ureaplasma infected infants. TNF-alpha immunoreactive cell density was very low in the gestational controls, but increased in both pneumonia groups. We conclude that persistent lung U. urealyticum infection may contribute to chronic inflammation and early fibrosis in the preterm lung. PMID- 11910509 TI - Aluminum phagocytosis in quadriceps muscle following vaccination in children: relationship to macrophagic myofasciitis. AB - Macrophagic myofasciitis (MMF) is a rare, seemingly emerging entity among adult patients in France. We encountered two children with the first two cases of MMF in North America. A 5-year-old male with chronic intestinal pseudo-obstruction required nighttime parenteral nutrition. Abnormal pupillary reflexes and urinary retention suggested a diffuse dysautonomia, which prompted a neurological diagnostic work-up. A 3-year-old child had developmental delay and hypotonia. Both children received age-appropriate immunizations. Quadriceps muscle biopsy from each child showed the typical patchy, cohesive centripetal infiltration of alpha-1-antitrypsin+, alpha-1-antichymotrypsin+, CD68+, PAS+, CD1a-, S-100-, factor XIII- granular macrophages with adjacent myofiber atrophy, dilated blood vessels, and mild endomysial and perimysial fibrosis. No myonecrosis was observed and no discrete granulomas were seen. A single aluminum peak was demonstrated on energy dispersive X-ray microanalysis. The etiology of the clinical symptoms in these cases and in cases reported as MMF remains intriguing. Despite numerous stains to demonstrate organisms, most infectious causes leading to macrophage activation were ruled out. These cases are being reported to increase awareness of this condition and to encourage a systematic epidemiologic and clinicopathologic study in North America. PMID- 11910510 TI - Maternal floor infarction and massive perivillous fibrin deposition: histological definitions, association with intrauterine fetal growth restriction, and risk of recurrence. AB - Maternal floor infarction (MFI) is a poorly understood placental lesion reportedly associated with intrauterine growth restriction (IUGR) and recurrence. In this study of MFI and the related placental disorder, massive perivillous fibrin deposition (MFD), semiquantitative histologic criteria for these diagnoses are defined and rates of IUGR and recurrence are assessed. Pathologic slides of 80 placentas diagnosed as MFI or MFD at the Brigham and Women's Hospital (1989 99) were reviewed and reclassified as classic MFI, transmural MFD, borderline MFD, or neither MFI or MFD. The prevalence of IUGR was determined, and placental slides from additional pregnancies were reviewed to evaluate recurrence. Among 25 cases originally diagnosed as MFI, 5 (20%) were reclassified by study criteria as classic MFI, 9 (36%) as transmural or borderline MFD, and 11 (44%) as neither lesion. Among 55 cases originally diagnosed as MFD, 27 (49%) were reclassified as transmural or borderline MFD, 4 (7%) as classic MFI, and 24 (44%) as neither lesion. IUGR was identified in no case with classic MFI, in 31% of cases with transmural or borderline MFD (P = 0.02), and in 11% of cases with neither lesion. Recurrence was documented in 1 of 7 (14%) second- or third-trimester placentas. Possible recurrence was suggested in 3 of 6 (50%) first-trimester spontaneous abortion specimens. Classification of intraplacental fibrin is subjective and problematic; almost half of potential cases of MFI or MFD did not fulfill our study's diagnostic criteria. MFD may be more common and more strongly associated with IUGR than classic MFI. Recurrence of these lesions appears to be infrequent among second- and third-trimester placentas, but may be relatively common in first-trimester spontaneous abortions. PMID- 11910511 TI - Peripheral nerve sheath tumors from patients with neurofibromatosis type 1 do not have the chromosomal translocation t(X;18). AB - Neurofibromatosis type 1 (NF1) is a common autosomal dominant genetic disorder that is caused by a mutation in the NF1 gene. Hallmark characteristics include dermal neurofibromas, cafe-au-lait spots, and learning disabilities. In approximately 25% of NF1 cases, plexiform neurofibromas, or peripheral nerve sheath tumors (PNSTs) that involve large segments of nerve sheath and nerve root, can form, of which a small percentage become malignant (MPNST). Most MPNSTs are composed of spindled neoplastic cells, and they can resemble other spindle-cell sarcomas, including leiomyosarcoma and monophasic synovial sarcoma. Histological diagnosis of MPNST is not always straightforward, and various immunohistochemical and molecular adjuncts can be critical in establishing a correct diagnosis. One example of genetic testing is the assay for the t(X;18) chromosomal translocation, which has been found to be common in synovial sarcomas. The aim of this study was to determine whether MPNSTs contain the t(X;18) chromosomal translocation. To detect the t(X;18) translocation product, SYT-SSX, total RNA was extracted from frozen archival tumors (15 dermal neurofibromas, 4 plexiform neurofibromas, and 7 MPNSTs) using Trizol. The RNA was then subjected to reverse transcriptase polymerase chain reaction (RT-PCR) to specifically amplify SYT-SSX. None of the dermal neurofibromas, plexiform neurofibromas, or MPNSTs analyzed were positive for SYT-SSX mRNA. The results indicate that the t(X;18) translocation is absent in neurofibromas and is not a marker for MPNST in patients with NF1. PMID- 11910512 TI - Unusual variant of holoprosencephaly in monosomy 13q. AB - The clinical phenotype related to the terminal deletion of the long arm of the chromosome 13 (the so-called 13q- syndrome) includes a considerable number of malformations, especially of the brain. This report describes five cases of a cerebral midline anomaly that leads to a particular clover-shaped type of holoprosencephaly in 13q- fetuses at different stages of the second and third trimesters of gestation. Our cases are compared to those in literature reviews. This malformation has only been described by computer tomography and magnetic resonance imaging in eight children of various ages and has been called "middle interhemispheric fusion" or syntelencephaly. Recently, the human gene ZIC2, the mutation of which leads to holoprosencephaly, has been mapped to the long arm of chromosome 13. on band q32. These findings suggest that this particular type of holoprosencephaly may be related to ZIC2 gene loss of function. PMID- 11910513 TI - Congenital nephrosis of the Finnish type: overview of placental pathology and literature review. AB - Congenital nephrosis of the Finnish type (CNF) is a rare, autosomal recessive disorder of glomerular filtration that results in massive proteinuria, edema, and ascites. Although previous studies describe the classic renal lesions characterizing this disorder, there are few documenting in detail the associated placental alterations. In this context, we present a case of CNF with emphasis on the placental pathology and compare our findings to what has been previously reported in the literature. A 36-year-old G2P1 with no significant medical history developed persistently elevated amniotic fluid alpha-fetoprotein in the absence of neural tube defects. Because of a clinical suspicion of CNF, she electively terminated the pregnancy at 19 weeks. Postmortem examination revealed characteristic renal changes, confirmed by electron microscopy, as well as significant placental villous edema. Although the placenta was not enlarged, the villi appeared profoundly hydropic. Extensive cystic vacuolar change was documented in both stem villi and tertiary villi, affecting 95% of the villi present. Since the fetus was not grossly edematous, the placental findings may represent the first sign of systemic hypoproteinemia. PMID- 11910514 TI - Apoptosis and white matter injury in preterm infants. AB - White matter injury in premature infants with or without intraventricular hemorrhage (IVH) remains an important cause of neonatal mortality and neurologic morbidity. The contribution of apoptosis to the cellular death in white matter injury in the preterm infant is unclear. The objective of this study was to determine whether apoptosis contributes to the cellular death in premature infants with cranial ultrasound (US) evidence of IVH and asymmetric periventricular echogenicity (PVE). Brain tissue incorporating frontoparietal white matter was obtained from 21 infants: 6 infants with severe IVH and asymmetric PVE (grade 1V IVH) on US (group 1); 9 infants with minimal IVH or normal US who died within 21 days (group II); and 6 infants with minimal IVH or normal US who died later (group III). The presence of DNA fragmentation, typical of apoptosis, was determined using a terminal deoxytransferase-mediated dUTD nick end labeling (TUNEL) assay. The TUNEL index for group I infants was significantly greater, i.e., 2.75 +/- 1.94% versus 0.84 +/- 0.70% for group II and 0.42 +/- 0.22 for group III infants (P = 0.004). Most cells showing reactivity had morphologic characteristics consistent with astrocytes and oligodendroglia. The number of white matter cells showing morphologic changes consistent with apoptosis, such as nuclear blebs and karyorrhexis, was also quantitated and was significantly more numerous in group I than in group II infants, i.e., 0.51 +/- 0.64% versus 0.02 +/- 0.05% (P = 0.0005), and group III infants, i.e., 0.10 +/- 0.18% (P = 0.03). These findings implicate apoptosis as a contributing mechanism for the cellular death in infants with IVH and asymmetric PVE. Strategies aimed at preventing the white matter injury will need to incorporate methods of inhibiting the ongoing process of apoptosis. PMID- 11910515 TI - Hormonal activity may predict aggressive behavior in neuroblastoma. AB - Overproduction of catecholamines (dopamine [DA], norepinephrine [NE]) and their metabolites (homovanillic [HVA] and vanillylmandelic [VMA] acids) characterizes neuroblastoma (NB). In previous studies, increased urinary DA/NE, and DA/VMA ratios have been associated with poor prognosis, whereas low DA/NE ratios have been associated with longer disease-free survival. Higher urinary VMA, HVA, and NE levels have been found in association with low MYCN amplification, in contrast to cases with high MYCN amplification in which normal levels have been found. It is then believed that an "immature" catecholamine pattern indicates poor prognosis. We correlated urinary DA, NE, VMA, and HVA levels with age, clinical tumor stage, histological features (favorable [FH]/unfavorable [UH]) and MYCN status of 33 patients with NB. DA/VMA and DA/HVA ratios were also calculated. Wilcoxon rank sum and chi-squared tests were performed to determine statistical significance. Eighty-eight percent (15/17) of stage 3-4 cases had DA levels >2 times the upper limit of normal, but only 8% (1/12) of stage 1-2 cases had DA levels twice the upper limit of normal. In 61% (11/18) of stage 3-4 cases, the VMA level was >10 times the upper limit of normal, in contrast to stage 1-2 cases, in which only one patient (1/15) had a VMA level >10 times the upper limit of normal. Similar findings were obtained with urinary HVA and NE. Patients older than 12 months of age at diagnosis also had higher urinary levels of DA, VMA, HVA, and NE than those of patients younger than 12 months of age at diagnosis. Eighty-two percent (14/17) of stage 3-4 cases had DA/VMA ratios <0.78, with the other 18% (3/17) showing ratios between 1.4 and 8.82 (all stage 4 and >12 months of age). In contrast, all stage 1-2 cases ((12)) had ratios <1.4. All (12/12) non MYCN-amplified cases had DA/VMA ratios <1.4 (0.06-0.84), while one MYCN amplified case (1/3) had a ratio of 8.82; the other two MYCN-amplified cases had DA/VMA ratios of 0.09-0.11. Twenty-nine percent (2/7) of cases with UH had a DA/VMA ratio >1.4, but in all FH cases (14/14) the DA/VMA ratio was <1.4 (0.08 0.084). Similar to previous studies, we found that aggressive NB is associated with higher urinary levels of DA, VMA, HVA, and NE. We also confirmed the previous observation that there appears to be a subset of NB in which a possible blockade in DA metabolism is associated with poor prognostic features (>12 months, stage 4, UH, and MYCN amplification). A seemingly novel observation in our study is that all high DA/HVA and DA/VMA ratios were obtained in stage 4 tumors, suggesting an association between the inability to metabolize DA and the acquisition of metastatic potential. On the basis of our results, we would like to emphasize the importance of determining not only DA, HVA, and VMA urinary levels, to support the diagnosis of NB, but also DA/HVA and DA/VMA ratios as a rapid initial assessment of prognosis in these patients. PMID- 11910516 TI - Neonatal alloimmune thrombocytopenia with HLA alloimmunization: case report with immunohematologic and placental findings. AB - Severe neonatal thrombocytopenia is associated with a significant risk of neonatal bleeding complications. It may result from increased consumption, increased destruction, deficient production, or abnormal sequestration within the spleen. When immune mediated, most cases of clinically significant neonatal thrombocytopenia are due to maternal alloimmunization to paternally derived platelet antigens present on fetal platelets. We present the clinical, placental, and immunohematologic findings of a case of severe neonatal alloimmune thrombocytopenia (NAITP) complicated by additional HLA group alloimmunization. The placenta showed chronic villitis of unknown etiology (VUE) and diffuse microthrombi within the villous capillaries, indicating that abnormal thrombogenesis can be a complication of severe NAITP. PMID- 11910517 TI - Cystic dysplasia of the rete testis: report of two cases and review of the literature. AB - Cystic dysplasia of the rete testis (CDT) is a rare congenital defect, characterized by multiple irregular cystic spaces in the mediastinum of the testis that may involve the whole gonad. A review of the literature has shown 32 reported cases, the majority of which were associated with ipsilateral renal malformations (agenesis/cystic dysplasia). Pathogenesis may be attributed to an early insult involving mesonephric duct development. Although treatment is surgical, when feasible, a conservative or nonoperative approach is suggested. Here we report two cases, one in a 3-year-old boy and one in a 10-day-old newborn. Concomitant cystic dysplasia of ipsilateral kidney was present in the former patient, while CDT was the solitary finding in the latter patient. Orchiectomy was performed in both patients, for extensive gonad involvement in the older boy and for suspected gonad torsion in the newborn patient. PMID- 11910518 TI - Gains of chromosome 8 in pleuropulmonary blastomas of childhood. PMID- 11910519 TI - The role of visual information in the control of scleral matrix biology in myopia. AB - Changes in eye size during the development of refractive error are accompanied by alterations in scleral biochemistry in both humans and animal models of myopia. This review discusses more recent data on scleral changes in mammalian models of myopia and considers the role of visual information in the control of scleral matrix biology. These visually-driven changes in scleral biochemistry are placed in the context of both the emmetropisation process and the abnormal enlargement of the eye that is characteristic of human high myopia. PMID- 11910520 TI - C5a receptor-mediated production of IL-8 by the human retinal pigment epithelial cell line, ARPE-19. AB - PURPOSE: C5a anaphylatoxin is a potent inflammatory mediator that acts on polymorphonuclear cells (PMN) and monocytes via C5a receptors (C5aR). It mediates chemotaxis of both cell types and stimulates cytokine release from the latter. To investigate whether C5a can act on retinal pigment epithelial (RPE) cells, we examined ARPE-19 cells for the presence of C5aR and the effect of C5a stimulation. METHODS: C5aR expression was measured by flow cytometry using specific anti-C5aR antibody and by RT-PCR analyses. Cells were stimulated with 50 nM C5a and the induction of IL-8 mRNA expression was measured by semi quantitative RT-PCR. RESULTS: Surface levels of C5aR on ARPE-19 cells were found to be comparable to those on human PMN. Stimulation with C5a induced a dose- and time-dependent increase of IL-8 mRNA expression. CONCLUSION: The findings of C5aR on ARPE-19 cells and induction of IL-8 mRNA upon C5a stimulation suggests that C5a may participate in the defense of choroidal and retinal tissue during inflammation. PMID- 11910521 TI - Platelet-activating factor (PAF) enhances apoptosis induced by ultraviolet radiation in corneal epithelial cells through cytochrome c-caspase activation. AB - PURPOSE: To examine the role of platelet-activating factor (PAF) on apoptosis of corneal epithelial cells exposed to radiation. METHODS: Rabbit corneal epithelial (RCE) and human corneal epithelial (HCE) cells were exposed to UVC radiation and then to carbamyl PAF (cPAF) for different increments of time. The PAF antagonist BN50739 was added 30 min before cPAF. The caspase inhibitors Ac-DEVD-CHO and Ac YVAD-CHO were added 1 h before, and the phospholipase A(2) (PLA(2)) inhibitor MAFP was added 3 h before UVC irradiation. FITC-dUTP TUNEL and DAPI staining were performed to assess the percentage of apoptotic cells. DNA ladder analysis was used to investigate apoptosis induced by different intensities of UVC (50-600 J/m(2)) with or without cPAF. Caspase activation and release of cytochrome c from mitochondria to cytosol were determined by Western blot. RESULTS: While only 2.7% of RCE cells were DAPI positive in controls incubated for 12 h, 44% of cells were stained positive 4 h after irradiation; these values increased to 63% in the presence of cPAF. Cells incubated with cPAF alone were similar to controls. TUNEL staining and DNA laddering showed also increased in apoptosis after PAF treatment of UV-irradiated cells and BN50739 blocked the effect of cPAF. cPAF increased caspase-3 activation induced by UV irradiation in HCE cells. Cytochrome c release from mitochondria to cytosol was observed 30 min after irradiation. cPAF almost doubled the release of cytochrome c at 30 min and 1 h. Here, too, BN50739 blocked the PAF effect. No release of cytochrome c by PAF was seen in non-irradiated cells, even at higher concentrations. MAFP caused a decrease in cytochrome c release from UV-treated cells, and caused an even greater inhibition of cytochrome c release in cells stimulated with PAF. CONCLUSIONS: PAF increases RCE and HCE apoptosis caused by UV irradiation by stimulating PLA(2), producing an early release of cytocrome c from mitochondria and activating caspase-3 by a receptor-mediated mechanism. This accelerating effect of PAF on the apoptotic cascade only occurred when corneal epithelial cells had been previously damaged by UV radiation. PMID- 11910522 TI - Long-term survival of allogeneic donor cell-derived corneal epithelium in limbal deficient rabbits. AB - PURPOSE: To investigate the capability of cultivated allogeneic epithelial stem cells to restore a functional ocular surface in a limbal deficient cornea; to verify the long term survival of epithelial allograft; and to examine the host immune response to heterologous cell transplant in a rabbit model. METHODS: Limbal deficiency was established by performing limbectomy on rabbits (n = 100). Corneal epithelial stem cells were obtained from the limbus and replicated in vitro without a supporting layer. The cell (3 x 10(5)) suspension was then transplanted via topical application as eye drops. Animals were divided into allograft, autograft, and control groups. Females were used as recipients and males as donors for the allograft. Corneas were collected at 7, 14, 21, 40 days as well as 2, 3, 7 and 8 months after cell transplantation. Experimental corneas were evaluated by histology, immunofluorescence, immunohistochemistry and Y chromosome analysis. RESULTS: A well-differentiated corneal epithelium was recognized at 14 to 40 days after cell transfer overlying an infiltrated corneal stroma. Corneal re-epitheliazation was confirmed in 31 of 36 allograft corneas. No significant immune rejection was noted. Stromal abnormality caused by previous limbal deficiency was mostly resolved three months after the regeneration of corneal epithelium. CONCLUSIONS: Transplanted corneal epithelial stem cells were able to differentiate into normal corneal epithelium in vivo without the use of membrane scaffolding. This non-autologous donor cell-derived corneal epithelium survived up to 8 months without immunosuppression and was able to reverse the stromal scarring. Thus, cultivated epithelial stem cells have great potential as an alternative to multiple-surgical procedures in the treatment of limbal deficiency states. PMID- 11910524 TI - Adrenergic supersensitivity of rabbit choroidal blood vessels after sympathetic denervation. AB - PURPOSE: To demonstrate the phenomenon of adrenergic denervation supersensitivity in rabbit choroidal blood vessels after superior cervical ganglionectomy. METHODS: Twenty four albino rabbits of both sexes weighing 2-3 kg were randomly separated into two groups. Twelve rabbits received bilateral superior cervical sympathectomy 2 weeks prior to the study (group s). The other 12 rabbits served as controls (group n). Four different concentrations of 0.1 ml phenylephrine, 0.05%, 0.025%, 0.013%, and 0.007% were slowly injected into the vitreous body near the retinal surface in group (n) and (s) rabbits (n = 6 in each group). The choroidal blood flow (PF), blood volume (CMBC), and velocity (V) were measured simultaneously by laser Doppler flowmetry (Perimed, PF 4001). RESULTS: The PF showed similar decreases in group (n) and (s) rabbits after injection of 0.05%, 0.025%, and 0.013% phenylephrine. With 0.007% phenylephrine, the PF remained unchanged in group (n) rabbits, but decreased significantly in group (s) rabbits (p = 0.0007). Velocity decreased similarly in both group (n) and (s) rabbits except for the 0.007% phenylephrine, concentration in which velocity decreased significantly in group s rabbits (p = 0.0001). There was no statistical difference in CMBC between group n and s rabbits at any of the test concentrations. CONCLUSIONS: The difference in PF decrease between group (n) and (s) rabbits with 0.007% phenylephrine demonstrated the existence of choroidal blood vessel denervation supersensitivity. The decrease in PF was achieved mainly through a decrease in blood cell velocity. PMID- 11910523 TI - Involvement of nitric oxide in the ocular hypotensive action of nipradilol. AB - PURPOSE: To evaluate the role of nitric oxide (NO) in the mechanism of the ocular hypotensive action of nipradilol, a beta-blocker with alpha( 1)-blocking activity. METHODS: Change in intraocular pressure (IOP) of albino rabbits was measured after a single application of carboxy-PTIO (c-PTIO), an NO trapping agent. Next, IOP change was measured every hour for 5 hours after the instillation of 0.25% nipradilol into one of the eyes with and without c-PTIO pretreatment of both eyes. IOP change induced by desnitro-nipradilol was also examined. The outflow facility and uveoscleral outflow were determined by two level constant pressure and anterior chamber perfusion methods before and at 3 hours after the application of nipradilol with and without c-PTIO pretreatment. RESULTS: Topical administration of c-PTIO showed no significant effect on IOP. Unilateral instillation of nipradilol reduced IOP significantly compared with control eyes with a maximum reduction of 3.6 mmHg and effect duration of 3 hours. Pretreatment with c-PTIO partially inhibited the reduction during an earlier period (1 approximately 2 hours) and completely at 3 hours. IOP change by desnitro-nipradilol was similar to that by nipradilol with c-PTIO pretreatment. Nipradilol increased both outflow facility and uveoscleral outflow at 3 hours, whereas pretreatment with c-PTIO inhibited both of these outflows. CONCLUSIONS: Results indicate that ocular hypotensive action by nipradilol during the relatively late period may be mainly due to enhancement of aqueous humor outflow by NO at least in the rabbits. PMID- 11910525 TI - Adenoviral vector-mediated beta-glucuronidase cDNA transfer to treat MPS VII RPE in vitro. AB - PURPOSE: To develop an effective therapy for treating glycosaminoglycan (GAG) storage in mucopolysaccharidosis VII (MPS VII) retinal pigment epithelium (RPE) in vitro using adenoviral vector mediated human beta-glucuronidase cDNA (Ad-GUSB) transfer. METHODS: Ad-GUSB was used to infect RPE at confluency. The transduction condition was optimized varying time of infection and number of infectious particles. The beta-glucuronidase (GUSB) activity was measured in transduced cells and media using a fluorogenic substrate. The GAG profiles were examined by metabolically labeling RPE with (35)Na(2)SO(4). RESULTS: Transduced RPE, irrespective of species or disease status, expressed a high level of beta glucuronidase. The expressed enzyme restored normal levels of GAGs in the RPE cells of homozygous affected MPS VII dogs by metabolizing stored GAGs. The over expressed enzyme (>10 000 nmoles/hr/mg) failed to restore normal level of GAGs. A high level of GUSB expression was maintained in vitro at least nine weeks. CONCLUSIONS: Adenoviral vector could mediate transfer of GUSB in MPS VII affected RPE and RPE of various species, and the expression was observed to be stable in vitro. However, controlled expression of GUSB was essential for the metabolism of stored GAGs to achieve normal levels. PMID- 11910526 TI - Vitreous levels of insulin-like growth factor-I in patients with proliferative diabetic retinopathy. AB - The levels of IGF-I in the vitreous body and the intraocular fluid of patients with proliferative diabetic retinopathy (PDR) were determined using radioimmunoassay (RIA). Eleven vitreous specimens were obtained from the intraocular fluid of eyes of patients with PDR who underwent surgery during the operation. Eleven intraocular fluids from the same patients during reoperations were compared with controls. The expression of IGF-I mRNA in cultured human Muller glial cells was evaluated using reverse transcription-polymerase chain reaction (RT-PCR). The mean IGF-I level in the vitreous samples during initial PDR surgery and reoperation was significantly higher than that found in the vitreous of the control (p < 0.05). The level of IGF-I increased in 6 of the 11 cases. Cultured human Muller cells expressed IGF-I mRNA. The results indicate increased levels of IGF-I both in the initial vitreous and ocular fluid at post operative re-proliferation. Muller cell is suggested as an origin of local IGF-I production. PMID- 11910527 TI - The effect of partial vitrectomy on blood-ocular barrier function in the rabbit. AB - PURPOSE: To compare ocular vascular permeability in the rabbit after vitrectomy as assessed by contrast-enhanced magnetic imaging (CE-MRI) and measurements of aqueous and vitreous humor protein concentration. METHODS: Partial vitrectomies were performed, irrigating with BSS or BSS PLUS. Post-operative vascular leakage was determined by CE-MRI following intravenous administration of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA). Aqueous and vitreous protein concentrations were quantified by standard biochemical assay. ERG evaluations were performed on postoperative days 1, 3, and 7. RESULTS: Using BSS as irrigant, breakdown of the inner blood-retinal barrier (BRB) occurred in 4/7 eyes on post operative day 1. The rate of Gd-DTPA leakage was significantly greater on postoperative day 1 than that in unoperated, control eyes, but declined approximately 50% by day 3. At both time points, outer BRB breakdown was restricted to the sclerotomy wounds. No BRB leakage was detectable in control eyes. Blood-aqueous barrier (BAB) leakage was bilateral on day 1. Significantly greater Gd-DTPA leakage occurred in the operated eye than in the nonsurgical contralateral eye. On day 3, approximately 40% bilateral reduction in leakage indicated resolution of BAB leakage. Notably, Gd-DTPA leakage of the BAB and BRB was significantly reduced in the BSS PLUS treated group. In contrast to MRI assessments, protein concentrations of the aqueous and vitreous in the surgical eye showed no detectable differences between BSS and BSS PLUS. Concurrent with the transient loss of ocular barrier function, ERG responses also declined. However, by day 7 greater than 90% recovery was noted in BSS PLUS treated animals but not in the BSS treatment group. CONCLUSIONS: CE-MRI is capable of detecting subtle changes in vascular permeability following ocular surgery. Advantages of using BSS PLUS compared to BSS as the irrigating solution can be detected using this technique. BSS PLUS's protection of barrier function is consistent with a rapid recovery in retinal function not observed in BSS treated eyes. PMID- 11910528 TI - Neuron specific enolase in retinal detachment. AB - PURPOSE: Neuron Specific Enolase (NSE) is released following central nervous system (CNS) distress. As retina is part of the CNS, NSE levels were measured in the subretinal fluid (SRF), aqueous, and serum of patients with primary rhegmatogenous retinal detachment (RD). METHODS: Radioimmunoassay was used to determine NSE levels in the SRF, aqueous, and serum of 13 patients (28-92 years old, mean = 71 years) with RD. As controls, NSE was measured in the aqueous of 6 patients undergoing cataract surgery and in serum of 18 patients without ophthalmological or neurological diseases. RESULTS: SRF levels of NSE ranged from 50-200 microg/l (mean +/- s.d. = 150 +/- 57). NSE levels in aqueous from patients with RD were 2-140 microg/l (mean +/- s.d. = 39 +/- 42), significantly higher than in controls (0-6 microg/l; mean +/- s.d. = 1.58 +/- 2.24; p = 0.04). Serum NSE levels in RD patients ranged from 6.5-80 microg/l (mean +/- s.d. = 26 +/- 21) and was significantly higher than in controls (5.3 +/- 1.66 microg/l; p = 0.005). CONCLUSIONS: Retinal neuron injury in retinal detachment (RD) releases sufficient Neuron Specific Enolase (NSE) to be detected in subretinal fluid, aqueous, and even in serum. Thus, NSE could index disease severity in RD and provide a means by which to assess the response to neuroprotection in RD. PMID- 11910529 TI - Hypoxia-induced [(3)H]D-aspartate release from isolated bovine retina: modulation by calcium-channel blockers and glutamatergic agonists and antagonists. AB - PURPOSE: The aim of the present study was two-fold: (a) to examine the effect of hypoxia on [(3)H]D-aspartate release from isolated bovine and human retinae, and (b) to investigate the regulation of hypoxia-induced neurotransmitter release by glutamate receptor agonists and antagonists. METHODS: Isolated neural retinae were incubated in oxygenated Krebs buffer solution containing [(3)H]D-aspartate and then prepared for studies of neurotransmitter release using the superfusion method. Release of [(3)H]D-aspartate was evoked by K(+) (50 mM) applied at 90 minutes (S(1)) and hypoxia (induced by exposure of tissues to solutions pregassed with 95%N(2): 5% CO(2) for 60 minutes) at 108 minutes (S(2)) after onset of superfusion. RESULTS: Under hypoxic conditions, pO(2) in normal Krebs buffer solution was reduced from 14.53 +/- 0.26 ppm (n = 6) to 0.54 +/- 0.04 ppm (n = 9) after one hour of gassing with 95% N(2): 5% CO( 2). Exposure to hypoxia elicited an overflow of [(3)H]D-aspartate yielding S(2)/S(1) ratios of 0.62 +/- 0.06 (n = 12) and 0.54 +/- 0.03 (n = 8) in bovine and human tissues respectively. In isolated bovine retinae, L- and N-calcium-channel antagonists diltiazem, nitrendipine, verapamil and omega-conotoxin significantly (p < 0.01 or higher) attenuated hypoxia-induced [(3)H]D-aspartate release. L-glutamate (30 microM) significantly (p < 0.001) potentiated hypoxia-induced [(3)H]D-aspartate release whereas kainate (30 microM) inhibited this response. NMDA (in concentrations up to 1 mM) had no effect on hypoxia-induced [(3)H]D-aspartate release. Antagonists of glutamate receptors and the polyamine site on the NMDA receptor inhibited hypoxia-induced release of [(3)H]D-aspartate in bovine retina with the following rank order of activity: ifenprodil congruent with MCPG > L-AP3 > MK-801. At an equimolar concentration (10 microM), L-AP3 but not ifenprodil, MCPG, MK 801 or arcaine, caused a significant (p < 0.001) inhibition of hypoxia-induced [(3)H]D aspartate release from human retinae. CONCLUSIONS: Hypoxia can induce the release of [( 3)H]D-aspartate from isolated bovine retinae by a calcium-dependent process. Hypoxia-induced [(3)H]D-aspartate release from isolated bovine retinae can be regulated by glutamate receptor agonists/antagonists and blockers of polyamine site on the NMDA receptor. PMID- 11910530 TI - Time-dependent changes in adrenal cortex ultrastructure and corticosterone levels after noise exposure in male rats. AB - In response to a stressful stimulus, there is a marked activation of the hypothalamic-pituitary-adrenal axis leading to a release of adrenocorticotropic hormone. This, in turn, acts on the zona fasciculata of the adrenal cortex to increase corticosterone plasma levels. Given the frequency of chronic intermittent noise exposure in man, we selected loud noise to evaluate concomitant changes in the ultrastructure of the adrenal cortex and corticosterone release. Following chronic (21 days, 6 h per day) loud white noise exposure (100 dBA, 0-26 KHz), we found the zona fasciculata to be most sensitive to time-dependent ultrastructural changes. These consisted of modifications in cell compartments involved in hormone synthesis and release. On the other hand, we found a progressive increase in corticosterone plasma levels which reached a plateau 9 days after noise exposure. The significance of these changes, in relation to phenomena like sensitization to repetitive stress, are discussed. Furthermore, the present data suggest that chronic loud noise exposure might potentially lead to endocrine dysfunctions. PMID- 11910531 TI - Distribution and changes with age of nitric oxide synthase-immunoreactive nerves of the rat urinary bladder, ureter and in lumbosacral sensory neurons. AB - In the distal parts of the urinary tract, nerves containing nitric oxide (NO) are either postganglionic parasympathetic nerves, with cell bodies in the major pelvic ganglia, or sensory nerves with cell bodies in the lumbosacral dorsal root ganglia. We have used indirect immunohistochemical techniques to examine the distribution and regional variation of nerves immunoreactive for neuronal nitric oxide synthase (NOS) in the urinary bladder, distal ureter and in neurons in lumbosacral dorsal root ganglia (L1-L2 & L6-S1) of young adult (3 months) and aged (24 months) male rats. Semi-quantitative estimations of nerve densities were made of NOS fibres innervating the dome, body and base of the urinary bladder and distal ureter. Quantitative studies were also used to examine the effects of age on the percentage of dorsal root ganglion neurons immunoreactive for NOS. The dome and the body regions, in both age groups, contained no NOS-immunoreactive axons. The bladder base and distal ureter in young adults showed sparse to moderate numbers of fibres immunoreactive to NOS within the urothelium and in the subepithelium and muscle coat. In the aged rat there were slight reductions in the densities of NOS-immunoreactive nerves in all three regions. In the lumbosacral dorsal root ganglia, the percentage of NOS-immunoreactive neuronal profiles showed a significant reduction from 4.6 +/- 0.2% in young adult to 2.7 +/- 0.2% (means +/- S.E.M) in aged rats. These findings suggest that the effects of NO on the bladder and distal ureteric musculature and also its expression in dorsal root ganglion neurons are affected in aged rats and that the micturition reflex may be perturbed as a result. PMID- 11910532 TI - A quick microwave histochemical stain for copper. AB - A rapid microwave method is described for staining copper in liver. This procedure was compared with a conventional method for copper. To this end, liver sections obtained from patients affected by several liver diseases associated with copper overload, were stained both with the standard rubeanic acid method for copper and with our modification of the same method, incorporating microwave treatment. Liver sections from a normal human newborn were used as a positive control. In Wilson's disease in the cirrhotic stage, copper was detected by the conventional method solely in periportal cells; following the microwave treatment, we were able to demonstrate copper in the whole lobule. In alcoholic cirrhosis, rubeanic acid stained copper only in a few periportal cells, while, by our modified method, copper was detected in almost all periportal hepatocytes. In chronic biliary tract disease, and in the newborn liver, copper was demonstrated in a few periportal cells by both the two histochemical procedures. In conclusion, although copper was detected by both procedures, a different degree of positivity was sometimes observed by using microwaves. Moreover, the microwave treated sections showed more contrast and less artifacts. From a practical point of view, for the simplicity of employment and, above all, for its quickness (10 min), we suggest the use of our method in all conditions where copper overload is suspected. PMID- 11910533 TI - Gene and protein expressions of type I collagen are regulated tissue-specifically in rat hyaline cartilages in vivo. AB - The present study was designed to investigate how rat hyaline cartilages at various sites in vivo express the gene and protein of type I collagen using in situ hybridization and immunohistochemistry. The gene of pro alpha 1(I) collagen was expressed by chondrocytes in articular cartilage, and the protein of type I collagen was identified in the cartilage matrix. In contrast, growth plate cartilage expressed the gene of pro alpha 1(I) collagen, but no protein of type I collagen. Neither gene nor protein of type I collagen was expressed in cartilages of trachea and nasal septum. The present study suggested that expression of type I collagen in hyaline cartilages may be regulated tissue-specifically at gene and/or protein levels. PMID- 11910534 TI - Phthalate esters influence FGF-2 translocation in Py1a rat osteoblasts. AB - Exposure of the Py1a rat osteoblastic cells to butyl benzyl phthalate (BBP) and dibutyl phthalate (DBP) showed that these endocrine disrupting chemicals (EDC) strongly and reversibly affect the cytoplasmic fibroblast growth factor-2 (FGF-2) translocation into the nucleus in a dose-dependent and time-related manner. Stimulation of cells with high concentrations of BBP or DBP for short timing gave results comparable to those of cells treated with low concentrations for long timing. By confocal laser scanning microscope (CLSM) analysis it was found that the first relevant effect resulted in an accumulation of FGF-2 near the nuclear envelope, sometimes in the shape of clusters; the growth factor was then translocated into the nucleus and, finally, after long periods of exposure, the basal nuclear and cytoplasmic binding, typical of unstimulated cells, was re established. In addition it was found that phthalate esters did not affect the FGF receptor 2 (FGFR-2) but decreased Con A binding indicating a possible inhibition of collagen fiber assembly. The different concentrations and timing of exposure of BBP and DBP affected the FGF-2 modulation in a similar way. Noticeable cumulative effects of BBP and DBP were not observed. PMID- 11910535 TI - Spatial distribution and cell kinetics of the glands in the human esophageal mucosa. AB - The glands in the human esophageal mucosa have been considered rudimentary, and have been poorly studied. Only recently, their role in the defense of the esophageal mucosa in gastroesophageal reflux disease has been put in evidence. In the present study, the presence of the different esophageal gland types was observed in 82 necropsy specimens. A precise topographical study was possible in 15 specimens. Cell proliferation parameters, namely DNA synthesis and mitotic index values in the squamous epithelium and in the esophageal glands were measured in 8 patients undergoing a surgical palliative esophagectomy, after in vivo labelling with bromodeoxyuridine. Upper mucosal, lower mucosal, and submucosal glands were observed in respectively 3.7%, 87% and 99% of the 82 esophageal specimens. Our topographical analysis showed that the upper mucosal glands, lower mucosal glands, and submucosal glands are occupying respectively 0.02%, 2.61%, and 3.96% of the esophageal surface. DNA synthesis and mitotic index values in the progenitor zone of the squamous epithelium were respectively 7.91% and 0.81%, whereas the proliferative activity in the glands was extremely low. The labelling index In the excretory ducts of the glands was only 0.07%, while no labelled cells nor mitotic figures were observed in any of the glandular acini. PMID- 11910536 TI - Methods on in situ hybridization, immunohistochemistry and beta-galactosidase reporter gene detection. PMID- 11910538 TI - Outcome from mild head injury in young children: a prospective study. AB - There is a lack of agreement regarding the long-term consequences of mild head injury (HI) at any age, with such effects rarely studied in early childhood. Given the rapid development occurring within the brain during this period, any disruption may have the potential to cause transient or permanent damage to brain structure and function. The present study sought to investigate the behavioral implications of such potential disruptions using a prospective, longitudinal design. Children aged 3-7 years at the time of injury, and suffering from mild HI, were evaluated acutely and at 6 and 30 months post-injury. Pre-injury data were collected with respect to communication, social skills, daily living skills and behavioral function. Results were compared to those from a non-injured control sample matched for age, gender, socioeconomic status, and pre-injury function. Findings showed few group differences. Children with mild HI performed similarly to controls on measures of intellectual ability, receptive language, and both everyday and spatial memory capacity. Group differences were identified for verbal fluency and story recall, with HI children failing to recover over time. PMID- 11910539 TI - Posttraumatic stress disorder and mild brain injury: controversies, causes and consequences. AB - Posttraumatic stress disorder (PTSD) is an anxiety condition that often arises following a traumatic experience. It has commonly been argued that impaired consciousness associated with mild brain injury (MBI) precludes encoding of the traumatic experience, and this prevents PTSD development. This review considers the available evidence on PTSD following MBI and indicates the distinctive nature of PTSD after MBI. The review then discusses the possible mechanisms that may mediate PTSD in this population. The interaction of PTSD and postconcussive symptoms is discussed within a cognitive model that emphasizes the role of catastrophic interpretations of postconcussive symptoms. Finally, the implications of PTSD after MBI for assessment and treatment are reviewed. PMID- 11910540 TI - Mild head injury: facts and artifacts. AB - While most would agree that mild traumatic brain injury (TBI) is associated with early neuropsychological problems, disagreement exists regarding their persistence and whether they are the cause of the disabilities experienced by some people. The aim of this study was to examine how the criteria used to define mild TBI and how the pre-injury characteristics of people affect their neuropsychological outcome. A total of 157 unselected hospitalized cases with Glasgow Coma Scale scores of 13-15 and 109 trauma controls were prospectively recruited and administered a number of cognitive measures at 1 month and 12 months after injury. The results indicated early impairments that decreased with time and the stringency of the definition of 'mild' TBI. The contribution of demographics was usually significant and often stronger than the mild TBI effect. Subtle variation of the demographics of the brain injured or the comparison subjects can be sufficient to mimic or mask mild brain injury effects. PMID- 11910541 TI - Cognitive outcome after mild and moderate traumatic brain injury in older adults. AB - This paper presents findings on the cognitive outcome of older adults sustaining mild traumatic brain injury (TBI). Results indicate that mild TBI patients who are 50 years or older, unlike those with moderate TBI, exhibit cognitive functioning that is comparable to noninjured controls by 1-2-months postinjury. However, these patients continue to report significant anxiety, depression, and somatic preoccupation despite their improvement on objective neuropsychological measures. The lowest postresuscitation Glasgow Coma Scale (GCS) score and the presence of intracranial pathology are more strongly associated with outcome than the durations of posttraumatic amnesia and impaired consciousness, possibly reflecting measurement issues in older persons who are likely to be injured in low velocity falls and to suffer delayed complications. A classification system that considers not only the GCS score but also the presence of intracranial pathology is sensitive to differences in the outcome of older adults, similar to the findings in young patients. The implications of these findings for older TBI patients and directions for research are discussed. PMID- 11910542 TI - Depression and posttraumatic stress disorder at three months after mild to moderate traumatic brain injury. AB - To investigate the frequency and risk factors of major depressive disorder (MDD) after mild to moderate traumatic brain injury (TBI), 69 TBI and 52 general trauma (GT) patients were prospectively recruited and studied at 3-months postinjury. There was a nonsignificant difference in the proportion of MDD patients in the TBI and GT groups. Therefore, a composite MDD group (TBI and GT patients) was compared to patients who were nondepressed. Female gender was related to MDD, but no other risk factors were identified. MDD was associated with disability (Glasgow Outcome Scale, Community Integration Questionnaire) and cognitive impairment. MDD was comorbid with posttraumatic stress disorder. Implications for postacute management of mild to moderate TBI are discussed. PMID- 11910543 TI - A dose-response relation of headers and concussions with cognitive impairment in professional soccer players. AB - The purpose of this study was to determine the effects of headers and concussions on cognitive impairment in professional soccer players. A group of 84 active professional soccer players from several premier league soccer clubs underwent neuropsychological evaluations. The dose-response relation between the number of headers in one professional season and the number of soccer-related concussions on cognitive functioning was investigated. It was found that the number of headers in one season was related to poorer results on tests measuring focused attention and visual/verbal memory. Soccer-related concussions were related to poorer results on tests measuring sustained attention and visuoperceptual processing. The findings suggest that headers as well as concussions separately contribute to cognitive impairment. PMID- 11910544 TI - Neuroimaging findings in mild traumatic brain injury. AB - The role of neuroimaging in the diagnosis and management of mild traumatic brain injury (TBI) is evolving. In general, the structural imaging techniques play a role in acute diagnosis and management, while the functional imaging techniques show promise for clarification of pathophysiology, symptom genesis, and mechanisms of recovery. A wide array of neuropathological processes are involved in mild TBI including changes in bone (e.g., a skull fracture), tissue density and water content (edema), blood flow, white matter integrity and pathway connectivity (diffuse axonal injury), and subtle changes in the neuronal and extracellular biochemical milieu. No single imaging technique is capable of addressing all these processes. It is, therefore, important to be aware of the advantages and limitations of the various available imaging modalities. This paper selectively reviews the pertinent literature on the structural and functional imaging in mild TBI. PMID- 11910546 TI - Assessment of response bias in mild head injury: beyond malingering tests. AB - The evaluation of response bias and malingering in the cases of mild head injury should not rely on a single test. Initial injury severity, typical neuropsychological test performance patterns, preexisting emotional stress or chronic social difficulties, history of previous neurological or psychiatric disorder, other system injuries sustained in the accident, preinjury alcohol abuse, and a propensity to attribute benign cognitive and somatic symptoms to a brain injury must be considered along with performances on specific measures of response bias. This article reviews empirically-supported tests and indices. Use of the likelihood ratio and other statistical indicators of diagnostic efficiency are demonstrated. Bayesian model averaging as a statistical technique to derive optimal prediction models is performed with a clinical data set. PMID- 11910545 TI - Postconcussional disorder following mild to moderate traumatic brain injury: anxiety, depression, and social support as risk factors and comorbidities. AB - Previous studies of postconcussional disorder (PCD) have utilized a dimensional approach (i.e., number and/or severity ratings of symptoms) to study postconcussional symptoms. This study used a syndromal approach (modified form of the DSM-IV criteria) for investigating risk factors for developing PCD, 3-months postinjury. The head trauma requirement was waived in order to determine specificity of symptoms to traumatic brain injury. Preliminary results from this ongoing study indicated significant risk factors including female gender, poor social support, and elevated self-reported depressive symptoms at 1-month postinjury. Comorbidities included concurrent diagnosis of major depressive disorder and/or posttraumatic stress disorder. Hispanics were significantly less likely to develop PCD than other racial/ethnic groups. PCD resulted more frequently from motor vehicle accidents and assaults. Screening tests for PCD risk factors/comorbidities performed shortly after injury (i.e., during routine follow-up clinic appointments) coupled with appropriate referrals for psychoeducational interventions and support groups may avoid prolonged loss of productivity and poor perceived quality of life in these patients. PMID- 11910547 TI - Treatment of post-concussion syndrome following mild head injury. AB - Approximately 38% of patients who sustain head trauma characterized by a brief disturbance of consciousness and clinically unremarkable neuroradiologic findings meet International Classification of Diseases 10th edition (ICD-10) diagnostic criteria for postconcussion syndrome (PCS). Physicians treat a majority of cases with nonsteroidal analgesics or antidepressants, and refer about 40% for psychological consultation. Psychological treatment typically involves education, reassurance, and reattribution of symptoms to benign causes. A review of controlled treatment outcome studies conducted over the past 2 decades in Scandinavia, Great Britain, Canada, and the United States suggests that early single session treatment can prevent the syndrome as effectively as traditional outpatient therapy. Several standardized, empirically supported treatment manuals are available. PMID- 11910548 TI - Prediction of outcome in mild to moderate head injury: a review. AB - This paper reviews the functional outcome of patients sustaining mild and moderate head injury (HI). Discrepancies across studies in the definition of minor, mild, and moderate HI are discussed in terms of hindering the interpretation of recovery. The predictive value of acute severity indices, neuroimaging findings, and the results of other techniques are summarized. Measurement of outcome based solely on the Glasgow Outcome Scale (GOS) is critiqued, and it is recommended that a differentiated outcome scale involving emotional, behavioral, cognitive, and physical domains should be used. PMID- 11910553 TI - Clinical characterization, linkage analysis, and PRPC8 mutation analysis of a family with autosomal dominant retinitis pigmentosa type 13 (RP13). AB - A Dutch family with autosomal dominant retinitis pigmentosa (adRP) displayed a phenotype characterized by an early age of onset, a diffuse loss of rod and cone sensitivity, and constricted visual fields (type I). One male showed a mild progression of the disease. Linkage analysis showed cosegregation of the genetic defect with markers from chromosome 17p13.1-p13.3, a region overlapping the RP13 locus. The critical interval of the RP locus as defined in this family was flanked by D17S926 and D17S786, with a maximal lod score of 4.2 (theta = 0.00) for marker D17S1529. Soon after the mapping of the underlying defect to the 17p13 region, a missense mutation (6970G>A; R2310K) was identified in exon 42 of the splicing factor gene PRPC8 in one patient of this family. Diagnostic restriction enzyme digestion of exon 42 amplified from genomic DNA of all family members revealed that the R2310K mutation segregated fully with the disease. The type I phenotype observed in this family is similar to that described for three other RP13 families with mutations in PRPC8. PMID- 11910554 TI - The epsilon4 allele of apolipoprotein E gene is a potential risk factor for the severity of macular edema in type 2 diabetic Mexican patients. AB - PURPOSE: The objective of this study was to determine the relationship between APOE polymorphism and the severity of retinal hard exudates in Mexican patients with type 2 diabetes. METHODS: We studied 36 patients with diabetic retinopathy and 22 unrelated and apparently healthy age-matched individuals. Blood samples for DNA genotyping and lipid profile were taken. Genotyping of polymorphic APOE alleles was done after polymerase chain reaction amplification of genomic DNA, digestion with HhaI, and agarose gel electrophoresis. Stereoscopic 35 degrees color fundus photographs were taken of seven standard fields. Diabetic retinopathy, macular edema and hard exudates were graded according to a standardized procedure. RESULTS: The results showed that the lipid profile was higher but not statistically significant (p > 0.05) in e4 allele carriers, with the exception of total lipids (p > 0.05). The frequency of severe retinal hard exudates was higher in those epsilon4 allele carriers (p < 0.05). The higher frequency of visual impairment (VA < 0.5 Log MAR) in epsilon4 carriers showed a tendency towards statistical significance (p = 0.057). CONCLUSION: Our results suggest that the epsilon4 allele of the ApoE gene is a potential risk factor for the severity of retinal hard exudates and visual loss in type 2 diabetic Mexican patients with diabetic retinopathy. PMID- 11910555 TI - Solitary retinal capillary hemangioma: lack of genetic evidence for von Hippel Lindau disease. AB - PURPOSE: To report the results of genetic testing for von Hippel-Lindau (VHL) disease in patients presenting with solitary retinal capillary hemangioma (RCH). METHODS: Ten patients with solitary RCH, who were excluded clinically as having VHL disease, underwent genetic testing using a combination of Southern blot, conformation sensitive gel electrophoresis, and direct sequence analysis. The results of the genetic tests were used to refine the empiric risk for VHL disease using principles of probability. RESULTS: Genetic testing for VHL disease was negative for mutation in all patients. The negative results of the genetic tests diminished the empiric risk for VHL disease by 100-fold. CONCLUSIONS: Solitary RCH can occur sporadically or be associated with VHL disease. In addition to clinical evaluation, genetic testing should be considered to exclude VHL disease with a high level of certainty. PMID- 11910556 TI - Progressive autosomal dominant optic atrophy and sensorineural hearing loss in a Turkish family. AB - PURPOSE: To describe the clinical features, mode of inheritance, and linkage analysis of ten affected members of a three-generation family with progressive optic atrophy and progressive hearing loss. MATERIALS AND METHODS: The proband, a 10-year-old boy, presented with progressive visual failure. Ten other members in his family, including his mother, half-sister, aunt, two uncles, grandfather, and some of the cousins, also had progressive visual loss and hearing loss. Six affected and four unaffected cases were examined in detail. Blood samples were drawn from 16 members for DNA extraction. Two loci previously described for optic atrophy were tested for linkage in the present family. RESULTS: The mode of inheritance was clearly autosomal dominant. Six members of the family were found to have progressive optic atrophy and hearing loss, both starting in the first decade of life. Total or red-green color blindness was detected in some patients. None of the members of this family showed evidence of other systemic disorders; however, four had blepharochalasis. No other cause could be found for the hearing or the visual loss. Linkage analysis excluded OPA1 and OPA2. CONCLUSION: The present Turkish family belongs to the group of individuals with autosomal dominantly inherited optic atrophies with hearing loss. Linkage analysis excluded OPA1 and OPA2, indicating that a novel gene defect underlies the disease in this family. Further genome-wide linkage analysis and identification of the disease associated gene will help define the pathophysiology of this syndrome. PMID- 11910557 TI - Colobomatous microphthalmia and orbital neuroglial cyst: case report. AB - We present the case of a boy with a congenital right orbital cyst with bilateral colobomatous microphthalmia. Neuroimaging studies excluded communication between the cyst and the eye and between the cyst and the central nervous system. Analysis of cyst fluid obtained by aspiration detected beta 2-transferrin by high resolution immunofixation (IFE). The cyst recurred two months following aspiration. It was then completely excised and histopathologic studies demonstrated a cyst containing neuroglial tissue. No recurrence was observed for 12 months following excision. PMID- 11910558 TI - A novel mutation in the FOXL2 gene in a patient with blepharophimosis syndrome: differential role of the polyalanine tract in the development of the ovary and the eyelid. AB - Blepharophimosis/ptosis/epicanthus inversus syndrome (BPES) is an autosomal dominant disorder characterized by abnormalities of the eyelids. We herein report a 12-year-old girl with BPES who had bilateral blepharophimosis, ptosis, hypertelorism, and downslanting palpebral fissures. Mutation analysis revealed the insertion of a cytosine (dup 1036C) within a wild-type run of six cytosines. A comparison of the phenotypic outcomes of the previously described mutations and the dup 1036C mutation reported herein suggest that the outcome is largely dependent on the involvement of the polyalanine tract (residues 221 to 231). We suggest that the polyalanine tract may have a differential role in eyelid and ovarian development and function. Further work is required to clarify whether ovarian function can be predicted on the basis of genotype. PMID- 11910560 TI - A computer-based register for inherited retinal dystrophies in Southern Africa. AB - A genetic register for inherited retinal degenerative disorders (RDDs) has been established at the Division of Human Genetics, UCT Medical School, Cape Town, South Africa. The primary role of the register is to monitor the progress of molecular research and to facilitate the efficient delivery of services, including genetic counselling, to respective family members and new patients. The database currently holds information on 1829 subjects. The RDD-specific breakdown of the data are presented. PMID- 11910559 TI - Visual improvement in Leber congenital amaurosis and the CRX genotype. AB - PURPOSE: In order to determine genotype-phenotype correlations in Leber congenital amaurosis (LCA), we analyzed the phenotype and genotype of 250 LCA children. We identified a heterozygous CRX mutation in an affected mother and son, and describe the ocular phenotype of the proband from birth through infancy to age 11 years. METHODS: Best-corrected Snellen visual acuities, electroretinograms (ERGs), and Goldmann visual fields were measured, while SSCP and direct sequencing were done for genotyping. RESULTS: The proband had congenital nystagmus, amaurotic, paradoxical pupils, and arteriolar narrowing, without a pigmentary retinopathy. The child had very poor fixation and wandering nystagmus at age 5 months, but had measurable vision at age 6 years. Snellen visual acuities were 20/900 at that time, and slowly improved to 20/150 by age 11 years. Perimetry revealed 60 degrees fields with the V4e target at ages 9 and 10 years, with a new 20 degrees inferior island to the III4e target. ERGs at 5 and 8 months were non-detectable, while the photopic ERGs at age 10 years and again at 11 years showed measurable cone a- and b-waves. At age 47, the phenotype of the affected mother consisted of hand motion vision, a pigmentary retinopathy, and non-detectable visual fields and ERGs. We identified a heterozygous CRX mutation, A177Delta1bp (529delG), in both affected individuals, which is predicted to cause a frameshift and introduces a premature termination codon at position 186. CONCLUSIONS: We report a CRX genotype with an ocular phenotype that consists of spontaneous, marked visual improvement in the proband from birth to age 11 years, which is unlike the previous six reports of LCA patients with the CRX genotype. PMID- 11910561 TI - Investigation of the prevalence of the myocilin Q368STOP mutation in Ugandan glaucoma patients. PMID- 11910564 TI - Identification of patients best suited for combined liver-kidney transplantation: part II. AB - Liver-kidney transplantation (LKT) should be reserved for those recipients with primary disease affecting both organs. However, increasing transplant list waiting times have increased the development and duration of acute renal failure before liver transplantation. Furthermore, the need for posttransplant calcineurin inhibitors can render healing from acute renal failure difficult. Because of the increasing requests for and controversy over the topic of a kidney with a liver transplant (OLT) when complete failure of the kidney is not known, the following article will review the impact of renal failure on liver transplant outcome, treatment of peri-OLT renal failure, rejection rates after LKT, survival after LKT, and information on renal histology and progression of disease into the beginnings of an algorithm for making a decision about combined LKT. PMID- 11910562 TI - A back migration from Asia to sub-Saharan Africa is supported by high-resolution analysis of human Y-chromosome haplotypes. AB - The variation of 77 biallelic sites located in the nonrecombining portion of the Y chromosome was examined in 608 male subjects from 22 African populations. This survey revealed a total of 37 binary haplotypes, which were combined with microsatellite polymorphism data to evaluate internal diversities and to estimate coalescence ages of the binary haplotypes. The majority of binary haplotypes showed a nonuniform distribution across the continent. Analysis of molecular variance detected a high level of interpopulation diversity (PhiST=0.342), which appears to be partially related to the geography (PhiCT=0.230). In sub-Saharan Africa, the recent spread of a set of haplotypes partially erased pre-existing diversity, but a high level of population (PhiST=0.332) and geographic (PhiCT=0.179) structuring persists. Correspondence analysis shows that three main clusters of populations can be identified: northern, eastern, and sub-Saharan Africans. Among the latter, the Khoisan, the Pygmies, and the northern Cameroonians are clearly distinct from a tight cluster formed by the Niger-Congo speaking populations from western, central western, and southern Africa. Phylogeographic analyses suggest that a large component of the present Khoisan gene pool is eastern African in origin and that Asia was the source of a back migration to sub-Saharan Africa. Haplogroup IX Y chromosomes appear to have been involved in such a migration, the traces of which can now be observed mostly in northern Cameroon. PMID- 11910565 TI - Living donor liver transplant recipients achieve relatively higher immunosuppressant blood levels than cadaveric recipients. AB - Two recent brief reports suggest that recipients of living donor liver transplants achieve higher levels of immunosuppressive agents than cadaveric (CAD) liver transplant recipients administered the same dose. These results could have important implications regarding the dosing of immunosuppressives in living donor liver transplant recipients. We report our findings relative to immunosuppressive doses and levels in a cohort of 46 living donor liver transplant recipients. Immunosuppressive blood levels and doses were recorded weeks 1, 2, 3, and 4 and months 2, 3, 4, 5, and 6 for 46 living donor liver transplant recipients and 66 matched CAD liver transplant recipients who underwent transplantation between August 1997 and May 2001. The ratio of level to dose also was recorded at each interval. The mean overall cyclosporine A dose was similar in living donor liver transplant recipients (323 mg/d) compared with CAD recipients (344 mg/d; P = not significant [NS]). The mean overall tacrolimus dose was 15% lower in patients who underwent living donor liver transplantation (LDLT; 5.7 mg/d) than CAD transplantation (6.7 mg/d), although statistical significance was not achieved (P =.08). The mean overall cyclosporine A level was 18% higher in those undergoing LDLT (275 ng/mL) than CAD transplantation (234 ng/mL; P =.015). The mean overall tacrolimus level was the same in living donor liver transplant recipients (10.8 ng/mL) and CAD recipients (10.2 ng/mL; P = NS). The overall cyclosporine A level-dose ratio was 26% higher for those undergoing LDLT (0.83) than CAD transplantation (0.66; P =.01). The overall tacrolimus level-dose ratio was 26% higher for those undergoing LDLT (1.82) than CAD transplantation (1.44; P =.01). In conclusion, (1) living donor liver transplant recipients achieve higher blood levels of tacrolimus and cyclosporine A for a given dose compared with CAD recipients, and (2) this difference is observed up to 6 months after transplantation, when hepatic regeneration is completed. PMID- 11910566 TI - Tacrolimus dosing requirements and concentrations in adult living donor liver transplant recipients. AB - Living donor liver transplantation in adult recipients is becoming increasingly common. The liver metabolizes most drugs, including immunosuppressive agents. Right-lobe grafts used in adult living donor liver transplantation consist of only 50% to 60% of the total liver. The purpose of this study is to determine whether there is a difference between tacrolimus doses and concentrations in patients who received a partial liver transplant from a living donor (LRD) versus those who received a whole-liver transplant from a cadaveric donor (CAD). Thirteen LRD recipients and 13 CAD recipients who underwent transplantation between April 1998 and July 2000 were included in this analysis. A CAD control group matched for age, sex, and race was used for comparison. Tacrolimus doses and concentrations were analyzed weekly for the first 4 weeks, then monthly for 6 months posttransplantation. There was no difference in acute rejection rates, renal and liver function test results, or number of potentially interacting medications administered between groups. LRD recipients required significantly lower doses of tacrolimus compared with CAD recipients at 2 weeks (0.058 v 0.110 mg/kg/d; P <.01), 3 weeks (0.068 v 0.123 mg/kg/d; P <.02), 4 weeks (0.086 v 0.141 mg/kg/d; P <.02), 2 months (0.097 v 0.141 mg/kg/d; P <.03), and 3 months (0.099 v 0.138 mg/kg/d; P <.03). Tacrolimus 12-hour trough concentrations were similar between groups at all times except for 2 weeks posttransplantation, when LRD recipients' concentrations were significantly greater than those of CAD recipients (12.4 v 9.5 ng/mL; P <.03). In addition, in the first month posttransplantation, LRD recipients were more likely to have greater concentrations of tacrolimus (>15 ng/mL; 22.1% v 9.2%; P <.01). In conclusion, LRD recipients have significantly decreased tacrolimus dosing requirements compared with CAD recipients during the first 3 months posttransplantation despite having similar tacrolimus concentrations. PMID- 11910567 TI - Influence of albumin supplementation on tacrolimus and cyclosporine therapy early after liver transplantation. AB - Liver transplant recipients administered gelatin (GEL) rather than human albumin solution (HAS) can become profoundly hypoalbuminemic in the early postoperative period and often have hepatic dysfunction at this time. The combined effect of these two abnormalities could be an increase in the unbound (active) concentration of low-extraction highly albumin-bound drugs, such as tacrolimus (TAC). This may increase the efficacy and/or toxicity of such drugs. We prospectively compared the clinical outcome of 69 de novo liver transplant recipients randomized primarily to TAC or cyclosporine (CYA) and secondarily to HAS or GEL therapy during the first 14 days after liver transplantation. Antipyrine clearance on the 7th postoperative day was used as a measure of liver metabolic function. Serum albumin levels were significantly lower in both GEL arms than HAS arms during the first 14 days (P <.001). Although antipyrine clearance was similar in all four trial arms, it was intermediate between that found in historic healthy controls and patients with cirrhosis (P <.0001). Serum creatinine concentrations were significantly greater in the TAC plus GEL arm than the other three arms (P <.001). The linearized treated acute rejection rate was significantly greater in the TAC plus HAS arm than the other three arms (relative risk, 2.02; 95% confidence interval, 1.07 to 3.78; P =.03). These data indicate that excess nephrotoxicity can occur with TAC in liver transplant recipients with impaired hepatic metabolism who are administered GEL. In addition, supplementary albumin may reduce the efficacy of TAC in liver transplant recipients at a time when the risk for rejection is greatest. PMID- 11910568 TI - Body surface area and body weight predict total liver volume in Western adults. AB - Computed tomography (CT) is used increasingly to measure liver volume in patients undergoing evaluation for transplantation or resection. This study is designed to determine a formula predicting total liver volume (TLV) based on body surface area (BSA) or body weight in Western adults. TLV was measured in 292 patients from four Western centers. Liver volumes were calculated from helical computed tomographic scans obtained for conditions unrelated to the hepatobiliary system. BSA was calculated based on height and weight. Each center used a different established method of three-dimensional volume reconstruction. Using regression analysis, measurements were compared, and formulas correlating BSA or body weight to TLV were established. A linear regression formula to estimate TLV based on BSA was obtained: TLV = -794.41 + 1,267.28 x BSA (square meters; r(2) = 0.46; P <.0001). A formula based on patient weight also was derived: TLV = 191.80 + 18.51 x weight (kilograms; r(2) = 0.49; P <.0001). The newly derived TLV formula based on BSA was compared with previously reported formulas. The application of a formula obtained from healthy Japanese individuals underestimated TLV. Two formulas derived from autopsy data for Western populations were similar to the newly derived BSA formula, with a slight overestimation of TLV. In conclusion, hepatic three-dimensional volume reconstruction based on helical CT predicts TLV based on BSA or body weight. The new formulas derived from this correlation should contribute to the estimation of TLV before liver transplantation or major hepatic resection. PMID- 11910569 TI - Right-lobe living related liver transplantation: evaluation of a comprehensive magnetic resonance imaging protocol for assessing potential donors. AB - The purpose of this study was to determine the practicability and diagnostic accuracy of a magnetic resonance (MR) protocol capable of replacing computed tomography, catheter angiography, and endoscopic retrograde cholangiopancreatography for the presurgical evaluation of potential liver donors before right hepatectomy. MR imaging (MRI) was performed on a 1.5 T scanner using a phased-array torso surface coil for signal reception. The following image sets were collected: axial two-dimensional (2D) T1-weighted fast low angle shot (FLASH), axial 2D T2-weighted half-Fourier acquisition single-shot turbo-spin echo (HASTE) with fat saturation, coronal MR cholangio-pancreatography (MRCP) based on 2D multisection HASTE and single-section single-shot rapid acquisition with relaxation enhancement (RARE) imaging, dynamic contrast-enhanced three dimensional (3D) FLASH, and contrast-enhanced T1-weighted FLASH. 3D FLASH data sets were collected before and after an intravenous administration of Multihance (gadobenate dimeglumine, Gd-BOPTA; Bracco, Milano, Italy), 0.2 mmol/kg of body weight. Thirty-eight potential liver donors were assessed by means of MRI. Twenty patients also underwent digital subtraction angiography (DSA). Of these, 16 patients underwent liver harvesting. MR angiography (MRA) data sets correlated with DSA results, and MRCP results correlated with intraoperative findings. Patients were excluded as potential donors based on insufficient liver mass of the left hepatic lobe (n = 5) or presence of hepatic pathological states (n = 9) seen at MRI, such as hemangiomas, focal nodular hyperplasias, or hepatic steatosis. MRCP showed the biliary system to the level of the first hepatic side branch. Dilated ducts were present in 4 patients. MRA depiction of hepatic arterial morphological characteristics correlated with catheter angiography results in all 20 patients: Three left hepatic arteries originating from the left gastric artery, three aberrant right hepatic arteries originating from the superior mesenteric artery, and two aberrant origins of both hepatic arteries and one common hepatic artery originating from the superior mesenteric artery were correctly identified on MRA. Similarly, the portal venous system was fully assessed on MRA. A comprehensive assessment of the hepatic parenchyma, biliary and pancreatic ductal system, and hepatic arterial, portal, and venous systems can be accomplished using the outlined protocol. PMID- 11910570 TI - Fatigue and physical function after orthotopic liver transplantation. AB - Over the last two decades, orthotopic liver transplantation (OLT) has become an established treatment for acute and chronic liver failure. OLT impacts not only on survival, but also on health-related quality of life. This study was undertaken to describe the self-rated health of Danish liver transplant recipients, compare their self-rated health against that of the general population, and to investigate associations between sex, age, diagnosis, time after OLT, and postoperative physical function and fatigue. All adult surviving liver transplant recipients who underwent OLT in Copenhagen, Denmark, from 1990 to 1998 (n = 154) were contacted by mail and asked to complete a self administered questionnaire. The questionnaire contained the 36-Item Short Form Health Survey, the Multidimensional Fatigue Inventory, the Hospital Anxiety and Depression Scale, and questions on marital status, education, and work. The response rate was 84.4% (n = 130). Liver transplant recipients reported poorer self-rated health than the general population in physical, but not in mental, health areas. One health aspect, fatigue, was investigated in great detail. This study found that liver transplant recipients experienced physical, rather than mental, fatigue. Diagnosis was found to be a predictor of postoperative physical function and fatigue because patients with an alcoholic or cryptogenic cirrhosis background had significantly poorer physical function and experienced more physical fatigue than liver transplant recipients with other diagnoses. Work status and survival time after OLT had significant effects on postoperative physical function and fatigue. Working and having undergone transplantation 4 to 5 years previously were associated with significantly better physical function and less physical fatigue than not working and having undergone transplantation 1 to 3 years previously. This study suggests that liver transplant recipients experience physical, rather than mental, impairment and fatigue and that diagnosis, work status, and survival time after OLT are associated with physical function and fatigue. PMID- 11910571 TI - Beyond the mean. PMID- 11910572 TI - Assessing health-related quality of life pre- and post-liver transplantation: a prospective multicenter study. AB - We report on a prospective multicenter study to assess pretransplantation and posttransplantation health-related quality of life (HRQL) of liver transplant recipients. HRQL was assessed at several timepoints using a self-completion questionnaire consisting of the EuroQol instrument (EQ-5D) and the 36-Item Short Form Health Survey (SF-36) health status profile measure. All individuals selected to receive treatment as part of the UK NHS liver transplantation program at each of six liver transplantation centers in England and Wales during a 2-year period (n = 542) were eligible to be included on the study. An overall response rate of 84% (455 responses) was achieved. A paired comparison of HRQL at listing and 3 months posttransplantation showed statistically significant improvements (P <.05) in all dimensions of the SF-36 (with the exception of Bodily Pain [P =.686]) and the EQ-5D tariff and visual analogue scale (VAS) scores. An analysis of posttransplantation HRQL over time for patients who survived until the end of the study (24 months posttransplantation) showed a statistically significant improvement (P <.05) for all dimensions of the SF-36 (apart from Mental Health [P =.245] and Role-Emotional dimensions [P =.265]) and the EQ-5D VAS and tariff scores. Adjusting for patients who died posttransplantation reduced mean EQ-5D tariff scores substantially, and the change over time in EQ-5D tariff scores was no longer statistically significant (P =.55). Results of regression analysis conducted to assess the importance of patient characteristics and center size on EQ-5D tariff and VAS scores generated posttransplantation indicated there was variation in scores according to patient age and center size. PMID- 11910573 TI - Predictors of mortality and stenosis after transjugular intrahepatic portosystemic shunt. AB - Transjugular intrahepatic portosystemic shunts (TIPSs) are used to treat variceal hemorrhage and refractory ascites. We sought to determine factors associated with stenosis and mortality after TIPS placement in patients with end-stage liver disease. This is a retrospective review of 90 TIPSs placed over a 3-year period. Demographic, clinical, and biochemical parameters were analyzed in univariate analyses to determine their association with stenosis and death. Multivariate analyses were conducted using logistic regression and Cox proportional hazard modeling. Thirty-five TIPSs were placed for recurrent variceal bleeding; 14 TIPSs, for uncontrolled variceal bleeding; 34 TIPSs, for refractory ascites; and 7 TIPSs, for other causes. The overall mortality rate was 33%, and 18 patients died within 30 days of TIPS placement. The 1-year stenosis rate was 49%. Fourteen patients underwent liver transplantation a mean of 116 +/- 143 days after TIPS placement. Prothrombin time greater than 17 seconds, serum creatinine level greater than 1.7 mg/dL, total bilirubin level greater than 3 mg/dL, and uncontrolled variceal bleeding as an indication for TIPS placement were significant predictors of 30-day mortality. Serum creatinine level was a predictor of 30-day mortality in individuals with recurrent variceal hemorrhage or ascites. Multivariate analyses showed that creatinine level greater than 1.7 mg/dL and uncontrolled variceal bleeding as an indication for TIPS placement were independently associated with 30-day mortality. Individuals with both coagulopathy and renal insufficiency had a 30-day mortality rate of 78%. Urgent placement of TIPS was associated with an increased risk for stenosis (hazard ratio = 4.5; 95% confidence interval, 1.9 to 10.1; P <.001), but no other clinical variables were associated with stenosis. Uncontrolled variceal bleeding as an indication for TIPS placement, coagulopathy, hyperbilirubinemia, and renal insufficiency were associated with increased mortality in patients with TIPSs. Individuals with both coagulopathy and renal insufficiency had high mortality. Urgent TIPS placement for uncontrolled variceal bleeding was associated with stenosis. PMID- 11910574 TI - Model for end-stage liver disease and Child-Turcotte-Pugh score as predictors of pretransplantation disease severity, posttransplantation outcome, and resource utilization in United Network for Organ Sharing status 2A patients. AB - The Model for End-Stage Liver Disease (MELD) has been proposed as a replacement for the Child-Turcotte-Pugh (CTP) classification to stratify patients for prioritization for orthotopic liver transplantation (OLT). Improved classification of patients with decompensated cirrhosis might allow timely OLT before the development of life-threatening complications, reducing the number of critically ill patients listed as United Network for Organ Sharing (UNOS) status 2A at the time of OLT. We compared the ability of the MELD and CTP scores to predict pre-OLT disease severity, as well as outcome and resource utilization post-OLT. Data from 42 consecutive UNOS status 2A patients undergoing OLT at a single center were used to calculate MELD and CTP scores at the time of status 2A listing. Multivariate analysis was used to determine the relationship between these scores and pre-OLT disease severity measures, survival post-OLT, and measures of resource use post-OLT. The MELD was superior to CTP score in predicting pre-OLT requirements for mechanical ventilation and dialysis. Neither score correlated with the resource utilization parameters studied. Only two patients died within 3 months post-OLT; neither score was predictive of survival in this cohort. In summary, the MELD is superior to CTP score in estimating pre OLT disease severity in UNOS status 2A patients and thus may help risk stratify status 2A or decompensated status 2B OLT candidates and optimize the timing of OLT. However, neither score correlated with resource use post-OLT in the strata of critically ill patients. PMID- 11910575 TI - De novo tumors after liver transplantation: a single-institution experience. AB - The aims of this analysis are to characterize the incidence and types of malignancies and tumor-specific mortality in our institution. Retransplantation, rejection episodes, and OKT3 use were evaluated. Our single-institution prospective database of 1,570 liver transplantations in 1,421 patients was analyzed. Data were statistically analyzed regarding sex, age at transplantation, time from transplantation to diagnosis of tumor, tumor type, and follow-up time. One hundred twenty-five patients (8.8%) developed de novo tumors; 69 patients were men, 56 patients were women. Seventeen patients received more than one allograft. De novo tumors were as follows: skin, 41; lymphomas, 35; lung, 11; colon, 6; anal, 2; rectal, 1; breast, 7; thyroid, 3; oropharyngeal squamous cell, 3; metastatic without primary tumor identified, 4; renal cell, 3; Kaposi's sarcoma, 1; angiosarcoma, 1; uterine, 1; ovarian, 1; pituitary, 1; pancreatic, 2; cholangiocarcinoma, 1; and esophageal, 1. These tumors developed in a statistically significant chronological sequence. Lung cancers and lymphomas showed shorter mean survival times, as well as greater mortality. OKT3 use and rejection did not show significance in tumor development. De novo tumors post liver transplantation affected our population in a distribution similar to that of the general non-transplantation population. Intense short courses of immunosuppression for rejection were not as important as chronic immunosuppression in the development of tumors. The risk for development was not enough to preclude transplantation. We found that tumors developed in chronological fashion. Therefore, directed surveillance, patient education, and early detection may facilitate earlier treatment. PMID- 11910576 TI - Intrahepatic cytokine profiles associated with posttransplantation hepatitis C virus-related liver injury. AB - Recurrent chronic hepatitis, cholestatic hepatitis, and acute rejection in conjunction with hepatitis C virus (HCV) recurrence are well-recognized clinical sequelae of reinfection of the hepatic allograft with HCV. The aim of this study is to characterize intrahepatic cytokine responses associated with reinfection of the allograft with HCV in these settings. Intrahepatic messenger RNA expression of T helper cell subtype 1 (TH1) cytokines interleukin-2 (IL-2), interferon gamma, and tumor necrosis factor-alpha and TH2 cytokines IL-4 and IL-10 was measured by real-time polymerase chain reaction system using TaqMan probes in 53 liver specimens from six groups of patients. These were: (1) recurrent chronic hepatitis C (CH-I; n = 15), (2) cholestatic hepatitis (n = 6), (3) acute rejection associated with HCV recurrence (AR-HCV; n = 12), (4) acute rejection in non-HCV-infected allografts (AR non-HCV; n = 5), (5) patients with chronic hepatitis C who did not undergo transplantation (CH-C; n = 10), and (6) non diseased liver tissue (n = 6). Intrahepatic viral loads were measured using an Amplicor monitor assay (Roche Diagnostic Systems, Branchburg, NJ). The CH-I and CH-C groups had similar TH1 intrahepatic cytokine profiles. Compared with the CH I group, the cholestatic group expressed increased levels of the TH2 cytokines IL 10 (P =.024) and IL-4 (P =.0024). The AR-HCV group also expressed more TH2 cytokines IL-10 (P =.014) and IL-4 (P =.034) compared with the CH-I group. Both the AR-HCV and AR non-HCV groups showed similar intrahepatic cytokine profiles. Intrahepatic viral loads were highest in the cholestatic group compared with the AR-HCV, CH-I, and CH-C groups (P =.0007). In conclusion, a novel observation is that the cholestatic group showed upregulation of the TH2 cytokines IL-10 and IL 4, in addition to high viral loads. In this setting, the TH2 immune response may favor viral replication and graft damage. PMID- 11910577 TI - Monitoring extracellular concentrations of lactate, glutamate, and glycerol by in vivo microdialysis in the brain during liver transplantation in acute liver failure. AB - Swelling of cerebral glial cells is a characteristic complication in patients with acute liver failure (ALF). This astrocyte edema may result in high intracranial pressure (ICP) and brain herniation before or during liver transplantation. Metabolic alterations responsible for the development of high ICP in patients with ALF are not fully understood. We describe changes in neurochemistry during liver transplantation using a cerebral microdialysis technique in a young man with severe ALF and cerebral edema. We found that the extracellular content of lactate ([lactate](ec)) gradually increased during the operation. Because cerebral oxygen saturation and [lactate](ec) to [pyruvate](ec) ratio were within normal limits, hypoxia was not likely to be responsible for the increased [lactate](ec) levels. Instead, we found that [lactate](ec) levels correlated in this patient with arterial lactate concentrations during and after grafting (r(2) = 0.96; P <.05), but did not correlate with arterial glucose concentrations (r(2) = 0.20; P = not significant). Also, [glutamate](ec) and [glycerol](ec) levels were severely elevated before liver transplantation, but tended to decrease in the hours after grafting. These findings indicate disturbances in glutamate neurotransmission, arachidonic acid metabolism, and lactate flux across the blood-brain barrier in patients with ALF. PMID- 11910578 TI - Imaging findings in invasive zygomycosis. PMID- 11910579 TI - Endoscopic surveillance and primary prophylaxis for upper gastrointestinal bleeding in liver transplant candidates. PMID- 11910580 TI - The descent of woman. PMID- 11910581 TI - The ERA trial: findings and implications for the future. PMID- 11910582 TI - Controversial issues in climacteric medicine I. Cardiovascular disease and hormone replacement therapy. International Menopause Society Expert Workshop. 13 16 October 2000, royal society of medicine, London, UK. AB - The clinical benefits of HRT are clearly established for the relief of menopausal symptoms, improving quality of life and the prevention of osteoporosis. Although research on the impact of HRT (oral, transdermal, tibolone, etc.) and on the effects of raloxifene on CVD is still ongoing, with certain unresolved controversies, studies using a variety of different HRT formulations have shown a clear benefit on surrogate markers of CHD and epidemiological and clinical, although not randomized, studies have demonstrated a CHD reduction in HRT-treated women. Today, HRT may be used for the primary prevention of CVD. Conversely, there is no clear reason to commence HRT solely or primarily to confer an immediate cardiovascular benefit in postmenopausal women with established CHD. Equally, there is no compelling evidence for discontinuing--or indeed not initiating--HRT in women without CVD because of concern about cardiovascular risk. In any case, all medical interventions should be individualized to the specific woman's age, characteristics and needs. The ultimate effects of different dosages, schedules and type of hormones used should be clarified, avoiding inferring the effects of one form of HRT to others. The importance of increased attention to life-style factors such as healthy diet, exercise and cessation of smoking should be underlined since these can confer specific benefits also to menopausal women. For women with known risks for CVD, HRT may contribute to the beneficial effects of life-style improvements and well established therapies (including blood pressure control, cholesterol-lowering drugs, aspirin, etc.). New strategies, including lower dosages, new estrogens, progestins, and new estrogen-like substances may be designed to target specific needs. PMID- 11910583 TI - Sociodemographic and clinical factors associated with HRT use in women attending menopause clinics in Italy. AB - OBJECTIVE: The aim of this analysis is to find any association between the use of hormone replacement therapy (HRT) and sociodemographic and clinical factors among women attending 54 menopause clinics in Italy. METHODS: The analysis includes data relating to 17,851 women who attended one of 54 menopause clinics in Italy for general gynecological evaluations and counselling between 1995 and 1997. The characteristics of women reporting ever HRT use were compared with those of never users. The odds ratios (ORs) of HRT use were computed in relation to selected reference categories, together with their 95% confidence intervals (CIs). RESULTS: Of the 17,851 women interviewed, 8539 reported ever HRT use. The mean age of the never and ever HRT users was 52.8 years and 53.7 years, respectively. Higher education was associated with an increased frequency of HRT use: in comparison with women reporting no or primary-/middle-school education, the OR of HRT use of women reporting a high-school diploma or university degree was 1.3 (95% CI 1.1-1.6). HRT use tended to be less frequently reported with increasing body mass index (BMI): in comparison with women whose BMI was < 23.4 kg/m2, the OR of HRT use in those with a BMI of 23.4-26.1 kg/m2 and > or = 26.2 kg/m2 was 0.8 (95% CI 0.8-0.9) and 0.6 (95% CI 0.5-0.7), respectively. Among the postmenopausal women, those who had undergone surgical menopause had an OR of HRT use of 1.3 (95% CI 1.2-1.4). A history of breast cancer was associated with a lower frequency of HRT use (OR 0.3, 95% CI 0.2-0.4). Likewise, a history of thromboembolic disease (OR 0.5, 95% CI 0.4-0.7) or myocardial infarction (OR 0.7, 95% CI 0.6-0.9) was associated with a lower frequency of HRT use. CONCLUSIONS: In our study population, the variable most closely related to HRT use was education/social class, thus underlining the impact of information campaigns on HRT and women's health. Among the medical determinants, our analysis indicates that a history of myocardial infarction, thromboembolic disease or breast cancer is associated with less frequent use of HRT. PMID- 11910584 TI - British-Asian women's views on and attitudes towards menopause and hormone replacement therapy. AB - OBJECTIVE: To establish an understanding of the views and values of Asian women living in the UK but of Indian subcontinent origin, in relation to the menopause and hormone replacement therapy (HRT): hopefully this will allow the development of strategies for increasing HRT uptake and long-term adherence. METHOD: This was a postal, structured questionnaire survey, completed anonymously by British-Asian women who expressed their opinions and knowledge about the menopause and HRT, including their views on service provision. RESULTS: Some 70% of women were in the age range 40-59 years; 74.5% had some education. In all 82% declared that they understood the menopause, and 77% thought that the menopause was a natural phenomenon. Over 75% were interested in seeking a medical opinion for management of the menopause. Of the women surveyed, 33% (n = 88) felt happy about and 46% (n = 123) felt frightened by the menopause. CONCLUSIONS: The results suggest an overwhelming demand for information and the need for special clinics run by female doctors who can communicate in the women's own language. Fears and concerns are similar to those of the Caucasian population. PMID- 11910585 TI - Dietary phytoestrogen intake in mid-life Australian-born women: relationship to health variables. AB - OBJECTIVES: To estimate the dietary intake of isoflavone-rich foods in a population-based cohort of Australia-born women, and to investigate whether a high intake of isoflavone-rich foods is associated with health-related variables. METHOD: The study considered a population-based cohort of Australian-born women aged 51-62 years, with interviews, blood and physical measurements taken in their own homes. Food frequency questionnaires included usual eating habits and isoflavone-rich foods. RESULTS: In total, 354 women (98%) returned both dietary questionnaires. Some 222 women (62%) reported consuming isoflavone-rich foods. Soy breads and milk provided the most servings per month. The mean intake of isoflavones calculated from soy beans, soy grits, tofu, soy milk, and soy and linseed bread in the whole cohort was 17 (standard deviation 35, range 0-340) mg/day. Fifty-one women (14%) consumed > 40 mg isoflavone/day. Compared with the rest of the cohort they had higher intakes of fruit, energy, protein, vitamins and minerals; ate a greater variety of vegetables; were more likely to exercise; were less likely to smoke; had lower mean body mass index, waist and hip measures and higher bone mineral density of the femoral neck; and had lower negative mood scores (all p < 0.05). CONCLUSION: Australian-born women in Melbourne have a wide range of intake of isoflavone-rich foods. For those with a high intake it seems to be one component of a healthier life-style. It is thus not possible to assign particular health benefits to the one dietary component. PMID- 11910586 TI - Effect of an estradiol gel with monthly or quarterly progestogen on menopausal symptoms and bleeding. AB - OBJECTIVE: To assess the acceptability, efficacy and endometrial safety of transdermal estradiol gel (Divigel/Sandrena) combined with monthly or quarterly oral progestogen (medroxyprogesterone acetate). METHODS: This 12-month, multicenter, open-label study was carried out at 12 study centers in Finland and Sweden. A total of 395 postmenopausal women received 1 mg estradiol in 1 g gel, daily, with oral medroxyprogesterone acetate 10 mg for the first 12 days every month (groups I and III) or every 3 months (group II). The main outcome measures were relief of climacteric symptoms, bleeding patterns and endometrial safety. RESULTS: All regimens reduced the severity of hot flushes, sweating episodes and vaginal dryness. In groups I and III, approximately 80% and 70% of women, respectively, had regular monthly withdrawal bleeding (excepting the first cycle), with irregular bleeding in 8.3% and 5.3% of treatment months. In group II, approximately 94% of women had regular tri-monthly withdrawal bleeding, with irregular bleeding in 10.7% of the treatment months. Endometrial hyperplasia was observed in 0.3% of women. More than 87% of subjects completed the study, and 97% of these rated the gel as acceptable or convenient. CONCLUSIONS: Both the 1- and 3-month regimens were equally effective in controlling climacteric symptoms and protecting against endometrial hyperstimulation. The bleeding patterns were comparable between groups and were similar to those reported for oral estrogens. Estradiol gel was highly acceptable to the majority of women. PMID- 11910587 TI - Comparison of the effects of continuous combined and sequential combined medroxyprogesterone acetate-estradiol treatment on the proliferation of MCF-7 cells. AB - OBJECTIVE: The aim of the study was to examine the effects of two different, clinically relevant, hormone replacement therapy (HRT) regimen types, continuous combined and sequential combined, on breast cancer cells by means of an in vitro model. The study was carried out using the C21-progestin medroxyprogesterone acetate (MPA) in combination with estradiol, and with the proliferation of MCF-7, a human breast cancer cell-line, as end-point. METHODS: Proliferation of MCF-7 cells was measured by means of a crystal violet staining technique. Growth was triggered using a constant estradiol concentration of 10(-10) mol/l, while varying the MPA concentration from 10(-11) to 10(-6) mol/l. RESULTS: The continuous combined model of treatment led to the inhibition of estradiol-induced growth of MCF-7 with MPA concentrations of 10(-10) mol/l and upwards, compared with estradiol-alone-induced growth. The sequential combined model showed a greater inhibition at the higher MPA concentrations of 10(-8)-10(-6) mol/l, with reduced sensitivity to inhibition at the lower MPA concentrations tested, of 10( 11)-10(-9) mol/l. The different treatment types resulted in significantly different sensitivities of the MCF-7 cells to inhibition of estradiol-induced proliferation at the higher MPA concentrations of 10(-8)-10(-6) mol/l. CONCLUSIONS: The results demonstrate the importance of considering in vivo factors in an in vitro model with regard to improving the cell culture techniques used, to obtain a clearer picture of the possible mechanisms involved in the potential breast cancer risk with different HRT regimens. PMID- 11910588 TI - Expression of adhesion molecules LFA-I and ICAM-I on osteoclast precursors during osteoclast differentiation and involvement of estrogen deficiency. AB - OBJECTIVES: Estrogen deficiency caused by the menopause or ovariectomy leads to stimulation of osteoclastogenesis. The adhesion molecules, leukocyte function associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1), are necessary for osteoclast formation. In this study, the expression of LFA-1 and ICAM-1 on osteoclast precursors during osteoclast differentiation, and the involvement of ovariectomy in the expression, were investigated. METHODS: Spleen cells isolated from normal or ovariectomized (OVX) mice were co-cultured with TMS14, stromal cells derived from mouse bone marrow, in the absence or presence of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) for 7 days. On days 3, 5 and 7 of culture, the expression of LFA-1 and ICAM-1 on osteoclast precursors was quantitated using indirect immunofluorescence and confocal laser cytometry, and, on day 7, the number of formed osteoclasts was measured by tartrate-resistant acid phosphatase (TRAP) stain. RESULTS: The level of ICAM-1 expression on osteoclast precursors gradually increased with osteoclast differentiation, whereas that of LFA-1 did not change. A high level of ICAM-1 was observed on the integrin beta 3-positive mononuclear cells. On the osteoclast precursors isolated from OVX mice, both the level of ICAM-1 expression per cell and the number of cells showing a high expression of ICAM-1 significantly increased, with an increase in the number of osteoclast-like cells. However, the level of LFA-1 did not change. CONCLUSIONS: These results indicate that the expression level of ICAM 1, but not that of LFA-1, is involved in osteoclast differentiation. Estrogen deficiency results in an increase in ICAM-1 expression on osteoclast precursors, which may be one of the mechanisms underlying bone loss following the menopause or ovariectomy. PMID- 11910589 TI - Problems of endometrial histology interpretation. PMID- 11910590 TI - (Greetings to Japan). PMID- 11910591 TI - The menopause in Japan--Konenki Jigoku. PMID- 11910592 TI - Does estrogen therapy enhance memory? PMID- 11910593 TI - Experience of menopausal symptoms by Chinese and Canadian women. AB - OBJECTIVE: This study examines the differences in frequency and distress caused by menopausal symptoms experienced by Canadian and Chinese women. DESIGN: Cross sectional surveys were conducted in Canada and China. SETTING: The Canadian studies were conducted in Toronto. The Chinese data were collected in Guangzhou City. PARTICIPANTS: Women, 47-62 years old, 2-7 years after a natural menopause, were recruited. METHODS: Two hundred and eighty-two Canadians were recruited for two menopause studies. Data from 297 Chinese were obtained through a household survey. From a 105-item symptom questionnaire, women indicated the frequency and distress caused by symptoms in the previous month. MAIN OUTCOME MEASURES: Frequency and mean distress score of problems were ranked and compared. Using the importance score method, a Chinese questionnaire was constructed and compared with the Menopause-Specific Quality of Life questionnaire. RESULTS: The frequency of symptoms and distress experienced by Canadian and Chinese women differed markedly. Fourteen of the 29 items differed between the Chinese questionnaire and the Menopause-Specific Quality of Life questionnaire. CONCLUSIONS: There are striking differences between Chinese and Canadian women in the frequency and distress caused by menopausal symptoms. The Canadian questionnaire may not be relevant for Chinese women. PMID- 11910594 TI - Lack of effects of transdermal estradiol on diastolic function: a randomized placebo-controlled double-blind short-term trial. AB - OBJECTIVE: Limited information is available on the effects of transdermal estradiol on diastole. The present study was a randomized double-blind placebo controlled trial designed to investigate the short-term effects of transdermal estradiol on left ventricular diastolic function in postmenopausal women. METHODS: The study included 45 women aged 50.8 +/- 3.6 years (25 randomized to the study group and 20 to placebo), who underwent Doppler echocardiography and determination of the plasma estradiol level after 4 and 8 weeks of transdermal estradiol administration in a dose of 50 micrograms per 24 h. RESULTS: There were no modifications in heart rate. Systolic blood pressure dropped in the study patients after 8 weeks (p < 0.03); diastolic blood pressure remained unchanged. Estradiol levels were 67 +/- 36 pg/ml at 4 weeks and 70 +/- 49 pg/ml at 8 weeks in the study group. Basal values of peak early and peak atrial velocities, acceleration time and rate, deceleration time and rate, early/atrial velocity ratio and pressure half-time were not significantly different between the estradiol and placebo groups. Doppler values remained unchanged after both 4 and 8 weeks in women receiving estradiol. Women with relatively high serum 17 beta estradiol levels (> 100 pg/ml) at 4 or 8 weeks of treatment did not present more pronounced changes in the Doppler-derived parameters compared with patients with low hormone levels. CONCLUSION: The results showed a lack of short-term effects of transdermal estradiol on left ventricular diastolic function in postmenopausal women, irrespective of serum estradiol levels. PMID- 11910595 TI - Concurrent use of simvastatin and estrogen--progestin therapy compared with each therapy alone for hypercholesterolemia in postmenopausal women. AB - OBJECTIVE: Substantial improvements in lipoprotein-lipid profiles have previously been shown with both simvastatin and combined estrogen-progestin therapy in postmenopausal hypercholesterolemic women. Since little is known about the impact of the concomitant use of these therapies, the effects of concurrent hormone therapy and simvastatin in hypercholesterolemic postmenopausal women have been evaluated. METHODS: Twenty-three postmenopausal women with fasting serum total cholesterol levels greater than 250 mg/dl received, in a randomized cross-over design, simvastatin (10 mg daily) for 8 weeks or postmenopausal hormone therapy (up to 1.25 mg of conjugated equine estrogens plus 5 mg of medroxyprogesterone acetate daily) for 8 weeks, with an 8-week wash-out interval between the two treatment periods. In a third, non-randomized treatment period after a second wash-out interval, each woman received a combination of simvastatin and postmenopausal hormone therapy in the same dosage regimens as above. Fasting blood was sampled monthly from baseline to measure total cholesterol, high- and low-density lipoprotein (HDL and LDL) cholesterol, triglycerides and lipoprotein(a). RESULTS: For total cholesterol, the mean decreases with hormone therapy, simvastatin and combination therapy were 12% (95% confidence interval 6 17%), 26% (20-31%) and 28% (24-31%), respectively, and for LDL cholesterol 21% (14-27%), 37% (30-44%) and 46% (41-51%), respectively. Simvastatin was more effective than hormone therapy (p < 0.001), while the effect of the combined therapy was even greater (total cholesterol, p = 0.012; LDL cholesterol, p < 0.001). The level of HDL cholesterol increased similarly with each treatment: 4% (-3-11%), 6% (2-10%) and 7% (2-13%), respectively. Triglyceride levels increased with hormone therapy and decreased with simvastatin (p < 0.001), while there was little change with the combination (effect of combined therapy vs. simvastatin, p = 0.002; vs. hormone therapy, p < 0.001). Both hormone therapy and combined therapy reduced lipoprotein(a) similarly (-23% and -14%, respectively, p = 0.078). Simvastatin had no effect on lipoprotein(a) levels. CONCLUSION: For postmenopausal women with hypercholesterolemia, use of a statin in combination with continuous combined oral estrogen and progestin therapy can result in a more cardioprotective lipoprotein-lipid profile than that achieved with either therapy used alone. PMID- 11910596 TI - Increased reduction in bone density and skin thickness in postmenopausal women taking long-term corticosteroid therapy: a suggested role for estrogen add-back therapy. AB - BACKGROUND: Long-term corticosteroid therapy is complicated by osteoporosis and generalized thinning of the skin. These two complications of such therapy were routinely assessed at the Menopause Clinic of St. Luke's Hospital Medical School, University of Malta. METHODS: A cross-sectional study was performed on 64 postmenopausal women who had been taking long-term corticosteroids. Each woman had her skin thickness measured using high-resolution ultrasound (22 MHz) and her bone density measured by dual-energy X-ray absorptiometry (DEXA). These measurements were compared with those of a control group (n = 557), a group of women who had sustained osteoporotic fractures (n = 180) and a group of women taking hormone replacement therapy (HRT) (n = 399). A longitudinal study of 29 postmenopausal women taking corticosteroids was also performed. This study compared results for women who, in addition to their corticosteroids, were taking HRT and for those who were taking corticosteroids alone. RESULTS: The cross sectional study showed that corticosteroid therapy was associated with the lowest mean skin thickness measurement (0.83 mm). Similarly, low mean bone density measurements for the lumbar spine (0.805 g/cm2) and left hip (0.715 g/cm2) were obtained for this group. The mean skin thicknesses in the control group and the HRT group were 0.93 mm and 0.935 mm, respectively, while that in the osteoporotic fracture group was 0.88 mm. The bone density of the fracture group was similar to that of the group of women taking long-term corticosteroids, with the lumbar spine having a mean density of 0.805 g/cm2 and 0.81 g/cm2, and the left hip having a density of 0.705 g/cm2 and 0.715 g/cm2, respectively. Bone densities were similar for the control group and the HRT group, and higher than that of the corticosteroid or fracture group. The lumbar spine had a mean density of 0.925 g/cm2 in the control group and 0.93 g/cm2 in the hormonally treated group. Both the treated and control groups had similar bone densities of the left hip at about 0.82 g/cm2. The small longitudinal study compared postmenopausal women on long-term corticosteroid therapy taking HRT with another group who were not taking HRT. This 4-year study revealed mean total increases in skin thickness of 6.1% and bone density of 5.5% (left hip) and 14.6% (lumbar spine) in the HRT group, since the start of the study. Conversely, the control group registered reductions over 4 years in both skin thickness (2.8%) and bone density (lumbar spine 4.5% and hip 5.0%). CONCLUSION: In postmenopausal women taking long-term corticosteroids, skin thickness and bone density were both decreased, but the addition of HRT as add-back improved the situation dramatically. Skin thickness and bone density in women taking long-term corticosteroids were comparable to those in women who had sustained osteoporotic fractures. It is therefore suggested that HRT be used as add-back therapy in postmenopausal women taking long-term corticosteroid therapy. PMID- 11910597 TI - Development and validation of a physical examination scale to assess vaginal atrophy and inflammation. AB - BACKGROUND: The assessment of vaginal atrophy is central to menopause-related research and clinical practice. However, methods of grading vaginal atrophy have not been subjected to reliability and validity assessments. OBJECTIVE: To assess the validity and reproducibility of selected vaginal examination findings proposed to represent vaginal atrophy and inflammation. DESIGN: Cross-sectional study. PARTICIPANTS: A convenience sample of 40 postmenopausal volunteers with past history of breast cancer. MEASUREMENTS: Participants completed questionnaires to assess vaginal symptoms. Pelvic examinations were carried out using specified criteria to assess the vaginal appearance. Vaginal cytological smears and pH measurements were performed using standardized techniques. RESULTS: A four-item physical examination atrophy scale had an alpha reliability of 0.48 and was statistically significantly correlated with pH (r = 0.55) and parabasal cells (r = 0.50). A three-item physical examination inflammation scale had an alpha reliability of 0.45. It was not significantly associated with vaginal smear measured inflammation. Masked inter-rater agreement for the examination was 90%. Self-reported symptoms of itching/irritation or dryness were poorly related to findings on physical examination. CONCLUSIONS: These results offer objective validation that the physical characteristics proposed to represent atrophy are related to biomarkers of this condition. A relationship between examination characteristics believed to represent inflammation and inflammation biomarkers was not upheld. PMID- 11910598 TI - Weight gain and the menopause: a 5-year prospective study. AB - OBJECTIVE: To investigate prospectively changes in weight, skin-fold measurements, waist circumference and waist/hip ratio in relation to changes in menopausal status, hormone therapy use and life-style factors. METHOD: The study was a 5-year follow-up of volunteers from a population-based cohort of Australian born women aged 46-57 years at baseline: 106 premenopausal, 106 perimenopausal and 21 hormone therapy users. RESULTS: Mean (SD) weight gain of the entire cohort over 5 years was 2.1 (5.1) kg. Baseline age was negatively associated with weight change (regression coefficient = -0.4, SE 0.1, p < 0.05). After 5 years, 20 women remained premenopausal, 80 were perimenopausal, 112 had become naturally postmenopausal and 21 remained on hormone therapy. Changes in weight were greater than zero (p < 0.05) in all groups except for the women who remained on hormone therapy. There was no significant difference in weight gain between women who remained premenopausal and those who had a natural menopause. Increases in suprailiac skin-fold measurements (p < 0.05) and in waist circumference and waist/hip ratio occurred in women who experienced the menopausal transition but not in those who took hormone therapy continuously. There was no association between weight change and baseline weight, exercise, alcohol intake or smoking. CONCLUSION: Weight gain was not related to change in menopausal status nor to any life-style factors measured. Women who were older at baseline gained less weight than the younger members. Suprailiac skin-fold measurements, waist circumference and waist/hip ratio all increased during the menopausal transition. Continuous hormone therapy users showed no gain in mean weight, suprailiac skin-fold measurements or waist measurements over the follow-up period. PMID- 11910599 TI - A general practice pilot audit study to assess advice and treatment offered to women following hysterectomy. AB - BACKGROUND: Current evidence suggests that information and treatment offered to women post-hysterectomy to prevent osteoporosis are poor. OBJECTIVES: To pilot a general practice audit protocol, to assess its ability to identify hysterectomized women and to offer appropriate information on fracture prevention. METHOD: A pilot audit study was designed to assess advice offered to hysterectomized women aged 25-64 years. Data on ovarian status were evaluated. Women were recalled for counselling if there was no evidence of advice on fracture prevention and if they were not taking hormone replacement therapy (HRT). Change in HRT usage was used as the outcome measure to assess the impact of the audit. RESULTS: Baseline data were collated for 5743 women. In 1456 (25%), both ovaries were removed and 4029 (70%) had one or both ovaries conserved; 258 (5%) had incomplete ovarian data. The average age at hysterectomy with both ovaries removed was 43.9 years, and with conservation of one or both ovaries was 40.5 years and 40.1 years, respectively. Some 2888 (50%) were ever-users of HRT (both ovaries removed 70%, one ovary removed 53%, both ovaries conserved 44%). A total of 2083 (36%) were known current users (both ovaries removed 53%, one ovary removed 40%, both ovaries conserved 30%). The mean duration of use in 2620 ever users was 44 months. This was similar for all the women irrespective of ovarian status. Eighty per cent of those given HRT reported receiving some advice. Seventy-seven per cent of those not given HRT had not received advice. By the end of the audit, 424 had commenced HRT (20% increase). Current use rose to 44% (both ovaries removed 59%, one ovary conserved 46%, both ovaries conserved 39%). Sixty three per cent of new users chose transdermal preparations. CONCLUSIONS: The mean age at hysterectomy of between 40.1 and 43.9 years indicates the potential for early ovarian failure. Use of HRT is associated with availability of counselling. Uptake was better than anticipated, but HRT usage was still well below optimum. This audit fulfills its objectives but not without cost implications. Year by year it should achieve significantly improved management and health of hysterectomized women, and improved standards of patient care. PMID- 11910600 TI - Migraine in a specialist menopause clinic. AB - OBJECTIVES: Epidemiological studies suggest that migraine and headache worsen during the climacteric. The authors noted that women attending a specialist hospital-based menopause clinic frequently reported vasomotor and other common climacteric symptoms but few spontaneously reported headache or migraine. The aim of this study was to assess the prevalence of migraine and headache in women attending this clinic. METHODS: Seventy-four women consecutively attending the menopause clinic at St. Bartholomew's Hospital were questioned about headache. Those with a positive response were further interviewed to obtain a headache diagnosis. RESULTS: Headache was found to be a common symptom affecting 57% of women in the 3 months before attending a specialist menopause clinic. Migraine affected 29% of patients in the preceding 3 months. This condition was associated with significant disability: 80% of women reported that attacks were more frequent than once a month; 75% reported that the attacks were severe; 50% reported that the duration of treated attacks was longer than 1 day. DISCUSSION: The high prevalence of headache and migraine in this group suggests that perimenopausal women should routinely be asked about headache and offered appropriate advice. This should include optimal attack therapy and strategies for preventing attacks, which may include hormone replacement therapy (HRT). Further studies are warranted to evaluate the relationship between climacteric symptoms, headaches, migraine and HRT. PMID- 11910601 TI - Hormone replacement therapy and venous thromboembolism. PMID- 11910602 TI - Discontinuation of HRT, memory disorder and cerebral blood flow. PMID- 11910603 TI - HRT and cancer again. PMID- 11910604 TI - HRT and breast cancer: is there any news? A clinician's perspective. PMID- 11910605 TI - Early follicular phase serum FSH as a function of age: the roles of inhibin B, inhibin A and estradiol. AB - OBJECTIVE: Reproductive aging in regularly cycling normal women is characterized by a gradual decline in ovarian follicle number and a progressive increase in serum follicle stimulating hormone (FSH), particularly over the age of 40 years. The lack of any consistent decrease in circulating estradiol and progesterone has led to the hypothesis that the FSH increase results from decreasing ovarian inhibin production. The aim of this study was to investigate the relationship between serum inhibins A and B, FSH and estradiol in normal women between the ages of 20 and 50 years. DESIGN AND PATIENTS: Serum from 66 regularly cycling subjects, aged 20-50 years, was collected on days 3-5 of the menstrual cycle for this cross-sectional study. MEASUREMENTS: Serum inhibin A and inhibin B levels were measured by specific enzyme-linked immunosorbent assays (ELISAs). Alpha subunit forms were determined by an immunofluorometric assay which detects all known monomeric and dimeric forms of inhibin A and inhibin B and free alpha subunit. FSH and estradiol levels were measured by immunoassay. Data were log transformed before analysis. RESULTS: Serum FSH, inhibin A and estradiol, but not inhibin B, were positively correlated (p < 0.05-p < 0.001) with age between years 20 and 50. Between 40 and 50 years, serum FSH was negatively correlated with inhibin B (r = -0.61, p < 0.001) and alpha subunit forms (r = -0.47, p < 0.05) and with estradiol (r = -0.39, p < 0.05), but not with inhibin A (r = -0.21, not significant). When log(FSH) was modelled as a function of log(inhibin B) and log(estradiol) with age fitted as a covariate, inhibin B only was a significant independent predictor of FSH (beta = -0.30, p < 0.01). Using purified inhibin A and B standards for the three assays, which were calibrated in terms of their alpha subunit content, serum inhibin A levels were 10-15% of those of inhibin B, with inhibin A + B levels being 22% of total alpha subunit levels. No significant correlation was observed between total inhibin alpha subunit and its dimers. The free alpha subunit, as determined from the difference in levels of total alpha subunit and inhibin A + B, remained relatively unchanged with age, suggesting that it is not differentially regulated. CONCLUSIONS: This study shows that, during the early follicular phase, FSH, inhibin A and estradiol but not inhibin B increase with age. Some of the increase in inhibin A and estradiol may be the result of accelerated follicular development with increasing age. Serum inhibin B and estradiol but not inhibin A are inversely correlated with FSH between ages 40 and 50, but only inhibin B is a significant independent predictor of FSH. This supports the postulate that inhibin B is the main form of inhibin regulating FSH at this stage of the menstrual cycle. During the early follicular phase, serum levels of inhibin A are presumably too low to play a significant physiological role or are less active. PMID- 11910606 TI - Cross-sectional study of determinants of menopausal age and hormone replacement therapy use in Italian women. AB - OBJECTIVE: The aim of the study was to analyze determinants of age at menopause and hormone replacement therapy (HRT) use. Women included were identified in a large cross-sectional study into epidemiology of the menopause among a sample of women visiting their general practitioners, conducted in Italy. METHODS: Eligible women were identified among consecutive patients, aged 44-66 years, who visited their general practitioner for a general health check-up during the period May November 1997. A total of 16,916 postmenopausal women were identified by 1123 general practitioners. RESULTS: Overall, the mean age at menopause was 50.2 (SD 3.8) years. Ever-married women reported a slightly higher age at spontaneous menopause than that of never-married women. The finding was significant, but the difference was small. Smoking women reported a younger age at menopause. No clear association emerged between age at menopause, physical activity and body mass index (BMI). A total of 3515 women (20.8%) reported HRT ever-use; the mean duration of use was 3.6 years. HRT use was more frequent among women of higher socioeconomic status, those with a lower BMI and smokers. In particular, in comparison with women reporting a BMI of < 25 kg/m2, the odds ratio (OR) of HRT use was 0.7 (95% confidence interval (CI) 0.7-0.8) and 0.5 (95% CI 0.4-0.6), respectively, in women with a BMI of 25-28 kg/m2 and > or = 29 kg/m2. No association emerged between alcohol consumption, level of physical activity and HRT use. Diabetic women reported HRT use less frequently than non-diabetic women. Likewise, hypertensive women, and those with a history of cardiovascular disease, were less likely to be HRT users than those not reporting these conditions. Women with a diagnosis of osteoporosis/osteopenia reported HRT use more frequently. CONCLUSION: This study, using a large dataset from an Italian population, has confirmed that smoking is related to age at menopause. It has also demonstrated that HRT is more frequently used by women of higher socioeconomic status, those with low BMI and smokers. Diabetes is associated with less frequent use of HRT; conversely, osteoporosis/osteopenia is associated with more frequent HRT use. PMID- 11910607 TI - Postmenopausal estrogen-progestin therapy and breast cancer: a clinical response to epidemiological reports. PMID- 11910608 TI - Psychosocial factors, attitude to menopause and symptoms in Swedish perimenopausal women. AB - OBJECTIVES: To analyze attitude to menopause from women's own accounts and to examine whether psychosocial factors and attitude are associated with symptom reporting. METHODS: The data form part of a population-based longitudinal study of a cohort of women who have been followed annually for 5 years using psychological interviews and rating scales, health screening and hormonal characterization. The data reported here were collected at the fourth follow-up when the women (n = 148) were 53 years old. RESULTS: Women were classified as perimenopausal (27%), postmenopausal (15%), hormone replacement therapy (HRT) users (52%) and hysterectomized (6%), based on self-reports. More than half the women (51%) had a positive attitude to menopause, 24% had a negative attitude and 25% had a neutral attitude. Menopausal status was not associated with attitude to menopause. Factor analysis of symptom ratings yielded ten independent factors comprising negative mood, vasomotor symptoms, decreased sexual desire, memory problems, sleep-related symptoms, vaginal dryness, urogenital problems, joint pain, vitality and increased sexual desire. Only vasomotor symptoms and joint pain were associated with menopausal status. The other symptoms were more strongly related to psychosocial factors, life-style and attitude to menopause. CONCLUSIONS: The results support the view of the menopause as a developmental phase associated with an increased self-awareness and a stronger personal identity. More than half the women held a positive view of the menopause, whereas the remaining proportion of women had either a negative or a neutral attitude. Only vasomotor symptoms and joint pain were associated with postmenopausal status. Other symptoms were significantly related to psychosocial factors, life style and attitude to menopause. PMID- 11910609 TI - Measurement-specific quality-of-life satisfaction during the menopause in an Arabian Gulf country. AB - OBJECTIVE: The aim of this study was to use an instrument, the menopause-specific quality-of-life satisfaction questionnaire for the postmenopausal period, in the United Arab Emirates (UAE). DESIGN: A cross-sectional descriptive study was used to generate menopause symptoms experienced by Arabian Gulf women. Measurement specific quality-of-life satisfaction questionnaires were used and face-to-face interviews were performed. SETTING: The study was based in primary health-care clinics in Al Ain City, Sharjah and Dubai Emirates, UAE. SUBJECTS: A multistage sampling design was used, and a representative sample of 450 UAE females aged 45 years and above were included during January-April 1999. RESULTS: Of the 450 women living in both urban and rural areas, 390 women agreed to participate (86.7%) and responded to the study. The mean age and standard deviation (SD) of the subjects was 56.5 +/- 6.6 years, and the median age of natural menopause in the present study was 48 years (mean +/- SD 48.4 +/- 3.8). The rate of consanguinous marriages in the sample was found to be 47.2%. The most common disease was found to be diabetes mellitus (10.3%), followed by osteoarthritis (7.7%), hypertension (7.2%) and asthma (6.2%), but the majority of subjects (68.7%) had no specific disease. Out of 29 possible symptoms, the mean number of symptoms was 7.57 (range 0-24). The most frequent symptom was 'aches in the back of the neck or head' at 46.4% followed by 'aches in the muscles/joints' at 34.6%. The least reported symptom was 'facial hair' at 15.9%. Increasing education resulted in more symptoms reported, and increasing parity resulted in fewer symptoms reported. In the present study, it was found that employed women experienced more symptoms and disorders. Of the total sample, 28.5% of the subjects reported no symptoms. In the four domains, 69% reported physical symptoms, 58.7% reported psychosocial symptoms, 40% reported vasomotor symptoms and 37.9% reported sexual symptoms. Spearman's rank correlation coefficient indicated that there is highly statistically significant concordance between the four domains (p < 0.01). CONCLUSION: The present study showed that menopause related symptoms in UAE women are fewer and of less severity than in Western women. The postmenopausal women, despite a continued decline in estrogen levels, reported few symptoms as part of a normal life stage, suggesting that they were able to cope with stress. PMID- 11910610 TI - The Menopause Rating Scale (MRS): comparison with Kupperman index and quality-of life scale SF-36. AB - OBJECTIVE: To evaluate further the Menopause Rating Scale (MRS) for scoring menopausal symptoms by comparison with other instruments relevant for women in their menopausal transition: the Kupperman index and the quality-of-life scale SF 36. METHOD: A population sample of 306 women, who had been randomly selected from an initially representative survey of German women (aged 40-60), completed three questionnaires in 1997: the Menopause Rating Scale (MRS), the Kupperman index and the short form-36 (SF-36). RESULTS: A comparison of the MRS with the Kupperman index produced a high correlation of raw scores (r = 0.91). The highest association of scores (80%) was found in the highest quartile of the MRS. The terms 'mild', 'moderate' and 'severe', relating to the degree of severity of menopausal symptoms, reflect different contents and spread in each scale, i.e. are not directly comparable. There is a strikingly good association between the subscales of the SF-36 and the MRS. The MRS correlates best with those dimensions of the SF-36 that are highly relevant for women in the menopausal transition. For this reason, the MRS can be utilized as an age- and condition-specific quality-of life instrument. CONCLUSIONS: The Menopause Rating Scale is a valuable modern tool for the assessment of menopausal complaints. It combines in practice excellent applicability and good reliability, and there are normal values for the population available. The MRS could serve as an adequate diagnostic instrument for menopausal quality of life. PMID- 11910611 TI - The Menopause Rating Scale (MRS): reliability of scores of menopausal complaints. AB - OBJECTIVE: To analyze the reliability of scores of the recently developed self administrative Menopause Rating Scale (MRS) in a follow-up investigation of a cohort after approximately one and a half years, and to look for possible reasons for variation. METHOD: A follow-up investigation of a random sample of an initially representative survey of German women (aged 40-60), which dated back to early 1996, was performed in late 1997. A subsample of 306 women participated. The MRS scale, a self-administrative standardized questionnaire, was applied with additional, mainly health-related, questions. RESULTS: The MRS results at baseline and follow-up were significantly correlated (r = 0.60). The majority of women remained in the category 'no or mild menopausal symptoms'. The kappa statistics showed significant agreement of the various subscales (total, somatic, psychological and urogenital scales) between the two measurements. Neither age nor social factors contributed to a change of score according to a multiple regression analysis. Some, but not all, health-related variables showed a slight association with change of score, such as satisfaction with health in general and, specifically, the presence of hypertension, cardiac and gastrointestinal diseases. No overall relation to hormone replacement therapy (HRT) was found during this observation period. CONCLUSIONS: The majority of women demonstrated sufficient reliability of MRS scores. Changes in the score after one and a half years were little influenced by the variables tested, except some health conditions such as cardiac disease. It should be stressed that the MRS has the benefit of being a self-administrative tool for the assessment of climacteric complaints with convenient applicability, and representative reference data have been collected in a German population. PMID- 11910612 TI - Which is the appropriate hormone replacement therapy after sub-total hysterectomy? PMID- 11910613 TI - Pulsatile release of sex steroids? A hypothesis to explain anomalies in hormonal therapy. PMID- 11910614 TI - Misleading claims for dietary isoflavones. PMID- 11910615 TI - How should we give progestogen? PMID- 11910616 TI - Effect of sequential transdermal progesterone cream on endometrium, bleeding pattern, and plasma progesterone and salivary progesterone levels in postmenopausal women. AB - BACKGROUND: Transdermal progesterone is being used in some countries as a purported treatment for menopausal symptoms, either alone or prescribed in conjunction with estrogen, but little information exists regarding the biological activity and effectiveness of this method of delivery of progesterone in protecting the endometrium from excess proliferation. This study was designed to evaluate the use of sequential transdermal progesterone. End-points evaluated included endometrial cellular response and bleeding pattern as well as plasma hormone levels and salivary progesterone estimations. METHOD: Twenty-seven postmenopausal women were treated with continuous transdermal estrogen (28-day cycle) and a cream containing 16, 32 or 64 mg of progesterone in each 4-cm extrusion from a tube of Pro-Feme administered daily in a sequential (days 15-28 of cycle) regimen. Blood and endometrial samples were analyzed for progesterone response prior to therapy, after the first 14 days of unopposed transdermal estrogen and following 14 days of transdermal progesterone. Saliva samples were taken during the last 14 days of the 84-day study, when the final progesterone cream therapy was being applied. RESULTS: Hormone assay indicated that physiological levels of estradiol were achieved, but progesterone levels were insufficient to induce any detectable change in the endometrium. Only one patient experienced bleeding during the study period. Levels of salivary progesterone were so variable as to be considered completely unreliable in determining the potential influence on biological activity. INTERPRETATION: Pro-Feme transdermal progesterone administered in a 16-, 32- or 64-mg daily dose for 14 days in a sequential regimen does not appear to be effective in inducing a secretory change in a proliferative endometrium. Salivary progesterone levels were not of value in managing the therapy of postmenopausal women. PMID- 11910617 TI - The effects of soy protein containing phytoestrogens on menopausal symptoms in postmenopausal women. AB - OBJECTIVE: To analyze the impact of soy protein dietary supplements containing phytoestrogens on menopausal symptoms in healthy postmenopausal women. METHODS: A double-blind, placebo-controlled trial was conducted in 94 healthy postmenopausal women aged 50-75 years, with 44 randomized to soy supplements containing 118 mg of isoflavones (daidzein, genistein, glycitein and their respective glycosides), and 50 to an identically presented casein placebo. A validated questionnaire on menopausal symptoms was administered at baseline and at 3 months of treatment. Compliance was assessed by high-performance liquid chromatography assay of urinary phytoestrogens. Statistical analysis was completed using non-parametric statistical methods and multivariate analysis. RESULTS: At baseline 80% of women recruited were experiencing menopausal symptoms, although symptom severity was mild. Those consuming phytoestrogen supplements had 13- and 17-fold increases in urinary excretion of genistein and daidzein, respectively, with no change in the placebo group. Active soy supplements did not significantly alter either individual symptoms or specific symptom category scores when compared to placebo. Within-group comparisons revealed that the active group reported a significant improvement in vaginal dryness (p = 0.01), libido (p = 0.009), facial hair (p = 0.04) and dry skin (p = 0.027). However, similarly, those on placebo reported an improvement in libido (p = 0.015), facial hair (p = 0.014) and dry skin (p = 0.011) but not vaginal dryness. CONCLUSIONS: In this group of 94 older postmenopausal women with a high frequency of mild menopausal symptoms, 3 months of soy supplements containing phytoestrogens did not provide symptomatic relief compared with placebo. PMID- 11910618 TI - Estradiol pharmacokinetics after transdermal application of patches to postmenopausal women: matrix versus reservoir patches. AB - OBJECTIVE: A new matrix 17 beta-estradiol transdermal patch incorporating lauric acid to improve estradiol skin absorption has been designed for hormone replacement therapy. Estradiol pharmacokinetics obtained with the prototype, its industrial counterpart, a matrix-type, System 50, and a reservoir-type, Estraderm TTS 50, transdermal patch have been compared. Each device delivers 50 micrograms estradiol daily. METHODS: Twenty postmenopausal women received each of the four formulations for 3 days in a Latin-square design and with a minimum 4-day wash out period between treatments. Estradiol plasma concentrations were measured by radioimmunoassay at 6, 12, 24, 48 and 72 h after application. RESULTS: The prototype patch and its industrial counterpart showed no significant difference in estradiol delivery, with 72-h systemic exposure to estradiol similar to that of the reservoir patch but greater than that of the reference matrix formulation, with average baseline-corrected concentrations (SEM) of 35 (4), 32 (3), 32 (2) and 19 (1.8) pg/ml, respectively. In addition, they ensured more stable delivery, with coefficients of variation of plasma estradiol concentrations (12-72 h) of 29, 41, 63 and 84%, respectively. All matrix patches demonstrated the same patients to be poor estradiol absorbers, different from those encountered with the reservoir patch type, despite an improved estradiol bioavailability with the lauric acid-containing matrix patch. CONCLUSION: Matrix patches incorporating lauric acid led to estradiol plasma levels more stable than with the reference matrix and reservoir patches, and greater than those with the reference matrix patch. PMID- 11910619 TI - Low-dose esterified estrogens (0.3 mg/day): long-term and short-term effects on menopausal symptoms and quality of life in postmenopausal women. AB - OBJECTIVE: To study the impact of low-dose unopposed esterified estrogens on menopausal symptoms and quality of life. METHODS: In a long-term, 2-year, randomized, double-blind, placebo-controlled study, 204 postmenopausal women were treated with esterified estrogens 0.3 mg daily or placebo. Menopausal symptoms were assessed with a modified Kupperman index at baseline, 3, 6 and thereafter every 6 months. In a second 12-week, open-label, short-term pilot study, 25 postmenopausal women with moderate to severe vasomotor symptoms were treated with esterified estrogens 0.3 mg daily for 12 weeks. Vasomotor symptoms and quality of life were assessed using the Greene scale and Quality of Life Menopause Scale (QUALMS). RESULTS: In the long-term study, significant (p < 0.05) reductions in total symptom scores were observed at each time point with esterified estrogens compared with placebo. Somatic symptom scores (hot flushes, night sweats, vaginal dryness) decreased significantly (p < 0.01) in patients treated with esterified estrogens 0.3 mg compared to baseline and placebo. In the short-term, open-label pilot study, the incidence of vasomotor symptoms was significantly (p < 0.01) reduced with esterified estrogens 0.3 mg from week 4 until the study end. Significant (p < 0.05) improvements versus baseline were seen in the somatic and vasomotor/sleep domains and in the total quality-of-life score. CONCLUSIONS: Esterified estrogens 0.3 mg given daily provide adequate menopausal symptom relief and improved quality of life in postmenopausal women. PMID- 11910620 TI - Retrospective self-report compared with menstrual diary data prospectively kept during the menopausal transition. AB - OBJECTIVE: To investigate the reliability of self-report of menstrual frequency and flow changes in the context of determining menopausal status categories, using data from the longitudinal phase of the Melbourne Women's Midlife Health Project (MWMHP). METHODS: Women reporting at interview at least one menstrual period during the previous 3 months are assigned pre- or perimenopausal status according to their responses to questions about changes in menstrual frequency and flow. For a sample of 72 such women, menstrual diary information was converted into standardized scores measuring change in frequency and flow of menses during the 2 years prior to interview. These scores, coded into categories, were used to derive measures of the sensitivity, specificity and predictive values of the interview responses. RESULTS: Self-report of change in menstrual frequency and flow have low sensitivity to measures based on prospectively kept menstrual diaries. CONCLUSIONS: Retrospective self-report at interview of changes in menstrual frequency and flow should not be regarded as reliable measures of actual changes in cycle parameters. PMID- 11910621 TI - Disturbances in postural balance are common in postmenopausal women with vasomotor symptoms. AB - OBJECTIVES: To establish the prevalence of unsteadiness and rotatory vertigo in peri- and postmenopausal women, and whether balance disturbances are more common in women with vasomotor symptoms and without hormone replacement therapy (HRT). METHOD: A validated questionnaire was sent to all 1523 women aged 54 or 55 years in Linkoping, Sweden. RESULTS: Daily or weekly unsteadiness was reported by 5%, and daily or weekly rotatory vertigo by 4% of all women. The frequency of vasomotor symptoms correlated with reported unsteadiness (rs = 0.23, p < 0.001). Fourteen per cent of women with daily vasomotor symptoms reported weekly or daily unsteadiness, compared with 3% of those without vasomotor symptoms (odds ratio (OR) 7.58, 95% confidence interval (CI) 3.72-15.45). The frequency of vasomotor symptoms correlated with rotatory vertigo (rs = 0.19, p < 0.001). Ten per cent of women with daily vasomotor symptoms reported weekly or daily rotatory vertigo, compared with 2% of women without vasomotor symptoms (OR 5.21, 95% CI 1.07 25.52). No correlation was seen between vasomotor symptoms and falls. Users of HRT had the same prevalence of balance disturbances as non-users. CONCLUSIONS: Women with frequent vasomotor symptoms seem to run a greater risk of unsteadiness and rotatory vertigo than do women without symptoms. This association may not be explained by means of a cross-sectional study, but there might exist a causal connection between vasomotor symptoms and balance disturbances. PMID- 11910623 TI - A business approach to dental practice. PMID- 11910622 TI - Intrauterine application of progestins in hormone replacement therapy: a review. AB - The intrauterine application of progestins as endometrial protection against hyperstimulation by estrogen replacement therapy has been investigated in clinical trials since the early 1990s and one product has become available for this indication. This review considers the available published and presented reports on intrauterine use of progestin to date. Reports of 19 studies were reviewed. These studies included both peri- and postmenopausal women (826 in total), treated with different types of estrogens administered via various routes. Progesterone was used in two small studies, while all other studies used different doses of levonorgestrel for periods ranging from 6 months to more than 5 years. Endometrial effects, bleeding profiles, systemic effects (symptoms and metabolic), as well as clinical experience, were considered and were comparable to other forms of continuous combined hormone replacement therapy (HRT). It is concluded that the current evidence supports complete endometrial protection and a good safety profile. The observed bleeding profiles appear favorable but have not yet been directly compared with other forms of continuous combined HRT. A favorable effect on serum lipids has been observed and also awaits direct comparative confirmation. Progestin-attributable side-effects, effects on bone and breast tissues and other systemic effects have not yet been studied. Acceptance by patients has been good, while insertion did not present undue problems for the investigating physicians. Retention of the studied intrauterine systems has been very good. Intrauterine use of progestins, especially levonorgestrel, by purpose-designed systems as part of combined HRT, is a new way of administration and carries good benefits, while some aspects require more clinical evidence. PMID- 11910624 TI - New guidelines and closer tabs on legislature aim for 1996. PMID- 11910625 TI - Hmmm ... something to think about.... PMID- 11910627 TI - The last diet. PMID- 11910626 TI - Keep moving--for a healthy life. PMID- 11910628 TI - Stress--a karate kick on the job. PMID- 11910630 TI - The challenge of coping with stress. PMID- 11910631 TI - Stress and dentistry. Better practice through control. PMID- 11910629 TI - Orthodontics and the mesially angulated molar. PMID- 11910632 TI - Hey, I didn't want that! PMID- 11910633 TI - Chaotic nightmare or fresh opportunity. PMID- 11910634 TI - Anyone have a match? PMID- 11910635 TI - Quality ... or quantity ... care? PMID- 11910636 TI - Is "managed care" an oxymoron? PMID- 11910637 TI - Freedom of Choice, who is it good for? PMID- 11910638 TI - Managed care: friend or foe? PMID- 11910639 TI - Direct reimbursement: an alternative employee benefit. PMID- 11910640 TI - The ABC's, PPO's and HMO's. PMID- 11910641 TI - Rapid, progressive facial swelling in a 23-year-old female. PMID- 11910642 TI - Oral lesions in Crohn's disease: review of the literature with case report. PMID- 11910644 TI - Another message from the heart. PMID- 11910643 TI - Stop to smell the roses. PMID- 11910645 TI - Working with geriatric dental patients. Maximizing function and functional dental health. PMID- 11910646 TI - Senior-Dent Program. PMID- 11910647 TI - Medical considerations for geriatric dentists. PMID- 11910648 TI - Growing old in a new dental age. PMID- 11910649 TI - Distinguished Service Award: Dr Mark Tajima. PMID- 11910650 TI - Effect of perimenopause on calcium absorption: a longitudinal study. AB - OBJECTIVE: Cross-sectional studies suggest that the rise in calcium requirement at the menopause may be attributable, at least in part, to a fall in intestinal calcium absorption. The aim of the present study was to determine the effect of the menopause on intestinal calcium absorption and the relationship between any change in calcium absorption and serum calcitriol. METHODS: Radiocalcium absorption and serum calcitriol were measured in 72 women aged 47.3 (standard error, SE 0.19) years who were initially premenopausal (as judged by menstrual history and serum follicle stimulating hormone (FSH)) and again 18 months later. RESULTS: Calcium absorption fell at the second visit from 0.72 (0.029)/h to 0.64 (0.029)/h (p = 0.003). Serum calcitriol had also fallen at the second visit from 124 (4.2) pmol/l to 111 (4.0) pmol/l (p = 0.007). At that visit, serum FSH exceeded the premenopausal reference range in 11 subjects and the menstrual cycle had become irregular in 24 of them. In the 11 women with raised FSH at the second visit, radiocalcium absorption fell from 0.85/h (0.097) at baseline to 0.57/h (0.049) (p = 0.008), but only from 0.70/h (0.028) to 0.65/h (0.033) (not significant) in the remaining 61. Similarly, radiocalcium absorption fell significantly (p = 0.003) in the 24 women with irregular menses, but not in the remaining 48 who continued to menstruate regularly. These changes in calcium absorption were still significant after correction for changes in calcitriol levels. CONCLUSION: The perimenopause is associated with a fall in calcium absorption, which is only in part attributable to a fall in calcitriol levels. PMID- 11910651 TI - Comparison of the efficacy and endometrial safety of two estradiol valerate/dienogest combinations and Kliogest for continuous combined hormone replacement therapy in postmenopausal women. AB - OBJECTIVE: To compare the efficacy and endometrial safety of two estradiol valerate/dienogest combinations with Kliogest in the treatment of postmenopausal symptoms. DESIGN: This was a double-blind, randomized, multicenter study. METHODS: Patients were randomized to estradiol valerate 2.0 mg/dienogest 2.0 mg (Climodien), estradiol valerate 2.0 mg/dienogest 3.0 mg (E2Val 2/DNG 3); or estradiol 2.0 mg/estriol 1.0 mg/norethisterone acetate 1.0 mg (Kliogest) once daily for 1 year. The primary efficacy variable was the Kupperman index. Endometrial safety was determined primarily by biopsy. RESULTS AND CONCLUSIONS: Climodien and E2Val 2/DNG 3 were therapeutically equivalent to Kliogest (mean changes in Kupperman index -20.1, -19.0 and -18.3, respectively). No statistically significant differences existed between treatment groups in the severity of postmenopausal symptoms. The incidences of endometrial atrophy were similar in all groups. Climodien appeared to be superior to Kliogest in terms of vaginal bleeding pattern, whereas E2Val 2/DNG 3 was associated with a slightly higher incidence and greater intensity of vaginal bleeding. The incidences of adverse events were similar in all groups. A greater proportion of women in the Kliogest and E2Val 2/DNG 3 groups experienced vaginal bleeding, whereas breast problems were more common with Climodien. Climodien and E2Val 2/DNG 3 induced desirable changes in insulin-like growth factor I (decrease) and sex hormone binding globulin (increase) that were not seen with Kliogest. PMID- 11910652 TI - Headache at menopause and in hormone replacement therapy users. AB - OBJECTIVES: To determine the prevalence of headache in menopausal women and to assess factors that may influence headache at menopause and in women using hormone replacement therapy (HRT). METHOD: A questionnaire survey was performed on 1000 consecutive women attending Leicester Royal Infirmary menopause clinics. Weight, blood pressure and smoking status were also recorded. RESULTS: Recurrent headache was common and reported by 850 women; 240 women reported a history of migraine. Most women (n = 617) had headaches more frequently than once monthly and 520 women had had at least one headache in the preceding week. Stress was the most common trigger factor (n = 704). A group of 'hormone-sensitive' women was identified (n = 259) whose headaches became worse at menopause and showed variable response to HRT. Logistic regression models showed reported history of migraine and more difficulty coping with stress to be strong predictors for worse headache at menopause and with HRT. Headache improved with age and increasing diastolic blood pressure. No other significant factors were identified. CONCLUSION: Headache is a substantial problem at menopause and in HRT users. It is difficult to predict which women will develop worse headaches at menopause and with HRT, but a history of migraine and reduced coping with stress were significant factors in this group. PMID- 11910653 TI - The role of megestrol acetate as an alternative to conventional hormone replacement therapy. AB - OBJECTIVE: To investigate the effect of megestrol acetate on menopausal symptoms, lipid metabolism, bone metabolism and coagulation. METHODS: In a prospective observational study, 71 postmenopausal women, for whom conventional hormone replacement therapy (HRT) was unsuitable, were treated with megestrol acetate 40 mg per day. At 0, 3, 6 and 12 months, fasting lipoproteins, bone biochemistry and thrombophilia profiles were measured and symptom score cards (Greene climacteric scale) completed. Bone mineral density measurement was performed at 0 and 12 months. RESULTS: Forty-one women completed the study. Treatment produced significant decreases in psychological (p < 0.001), vasomotor (p < 0.001) and somatic (p < 0.01) symptoms. There were significant reductions in triglycerides (p < 0.001), total cholesterol (p < 0.001), very-low-density lipoprotein (VLDL) (p < 0.05), low-density lipoprotein (LDL) (p < 0.001), high-density lipoprotein (HDL) cholesterol (p < 0.001) and lipoprotein(a) (p < 0.001). Levels of protein C were reduced (p < 0.05) and fibrinogen increased (p < 0.05). Protein S, plasminogen and antithrombin III levels showed an upward trend, which did not reach statistical significance. Biochemical markers of bone turnover did not change, apart from a significant decrease in alkaline phosphatase. Spinal bone density decreased significantly after 12 months, while femoral neck density remained unchanged. CONCLUSIONS: Megestrol acetate controls menopausal symptoms, has equivocal effects on cardiovascular risk markers and does not increase bone density. It is useful where estrogen is contraindicated. PMID- 11910655 TI - Advertising standards for complimentary medicines at the climacteric. PMID- 11910654 TI - Menopausal symptoms: experience of Chinese women. AB - OBJECTIVES: To investigate the prevalence of symptoms in middle-aged Chinese rural and city women of different occupations; and to explore the relationship between symptoms, hormone levels and other factors. METHODS: This cross-sectional study included 806 women aged 41-60 years, selected by multistage cluster sampling and a structured interview questionnaire. The response rate was 95%. The sample compromised 402 professional urban women and 404 women farmers living in rural areas. Some 209 women were randomly selected from the two groups for hormonal assay and bone mineral density screening. RESULTS: The professional group was more symptomatic than the farming group (p < 0.01). The presence of symptoms was significantly related to an increasing level of education. There were no significant differences between occupational groups in levels of estradiol follicle stimulating hormone (FSH) and luteinizing hormone (LH). Estradiol level and bone mineral density were decreased and FSH and LH increased in the postmenopausal group (p < 0.05) compared with the other menopausal groups. Hot flushes were the only symptom to be significantly associated with hormone levels (estradiol and LH). The occurrence of symptoms was significantly related to indications of bone and joint disease, heart disease, primary dysmenorrhea, decline in sexual interest, irregular menses and feelings of becoming older, sad and lost. These factors were all reported more often by the professional women than by the farmers. CONCLUSION: Symptom experience in mid-life Chinese women is related to both biological and psychosocial factors. PMID- 11910656 TI - Vitamin D receptor and estrogen receptor gene polymorphisms in postmenopausal Danish women: no relation to bone markers or serum lipoproteins. AB - OBJECTIVE: To investigate the polymorphisms of the vitamin D receptor (VDR) and estrogen receptor (ER) genes in relation to biochemical markers of bone turnover (serum osteocalcin and urinary collagen type I degradation products (CrossLaps), and to study ER genotypes in relation to serum lipoproteins, blood pressure, or changes in these parameters after 2 years of hormone replacement therapy (HRT) in 499 Danish postmenopausal women. METHODS: The VDR gene polymorphisms were determined by means of the three restriction enzymes, i.e. BsmI, ApaI and TaqI, while the ER gene polymorphisms were determined by means of the PvuII and XbaI restriction enzymes. Serum osteocalcin, urinary CrossLaps and the lipoproteins were also assessed. Body mass index was recorded. RESULTS: The VDR or ER genotypes did not differ significantly with respect to age, age at menopause or body mass index. No significant effect of VDR or ER genotype on bone turnover was found. Furthermore, we were unable to find any relationship between ER genotype and lipoproteins or blood pressure at baseline, or changes in these parameters during HRT. CONCLUSION: A clinically significant relationship between VDR and ER genotypes and biochemical markers of bone turnover or serum lipoproteins could not be demonstrated in healthy Danish postmenopausal women. PMID- 11910658 TI - Significant differences in estradiol bioavailability from two similarly labelled estradiol matrix transdermal systems. AB - OBJECTIVE: To compare the bioavailabilities of estradiol delivered by two transdermal estradiol matrix systems; Alora and Evorel. STUDY DESIGN: A single center, open-label, randomized, two-period cross-over study in 33 postmenopausal women. The subjects received two successive 84-h applications of either Alora or Evorel (each labelled to deliver 50 micrograms/day 17 beta-estradiol) in a randomized sequence. Serial serum samples, collected over the 84-h period following the application of the second patch, were analyzed for estradiol using a validated radioimmunoassay method. RESULTS: The fluctuation index produced by Evorel was significantly higher than that produced by Alora (Evorel, 135%; Alora, 76%; p < 0.0005). In addition, the estradiol baseline-corrected area under the curve for Evorel was significantly lower than that for Alora (Alora, 2871.8 pg h/ml; Evorel, 1870.6 pg h/ml; p < 0.0005). Both patches were found to be generally well tolerated. CONCLUSION: Alora delivered a higher, more consistent concentration of estradiol into the systemic circulation over the entire dosing interval than did Everol. Although the full clinical significance of these findings is currently unknown, this study demonstrates that there are significant differences in estradiol delivery from these two products, although they are labelled with the same nominal delivery rate. PMID- 11910657 TI - Comparison of different treatment modalities for postmenopausal patients with osteopenia: hormone replacement therapy, calcitonin and clodronate. AB - OBJECTIVE: To evaluate the effectiveness of hormone replacement therapy (HRT), clodronate, calcitonin and a clodronate plus calcitonin combination in postmenopausal patients with osteopenia. METHODS: One hundred postmenopausal patients with osteopenia, with bone mineral density (BMD) measurements at least one standard deviation below the mean value for young premenopausal subjects (T score < -1), were studied. They had no contraindications to HRT, clodronate or calcitonin use and were randomized to four different treatment groups. Patients in group I were treated with transdermal estradiol 50 micrograms/day and oral medroxyprogesterone acetate 10 mg/day during the last 12 days of the month; group II received oral clodronate 400 mg/day for 1 month out of every 3 months; group III received calcitonin nasal spray 100 IU/day; and patients in group IV were treated with oral clodronate 400 mg/day for 1 month out of every 3 months plus calcitonin nasal spray 100 IU/day. Elementary calcium 1000 mg/day was supplemented to patients in all groups. Spinal and femoral neck BMD measurements and markers of bone mineral metabolism were measured in each patient before treatment and 6, 12 and 18 months after treatment in 86 patients. RESULTS: Significant increases in mean lumbar spine BMD were found in the group receiving HRT, and at the end of 18 months there was a 2.69 +/- 0.76% increase, compared with baseline. Mean femoral neck BMD also increased by 2.22 +/- 0.57% in the HRT group; this was significantly different from baseline, resulting in a higher bone mass gain than in the other three groups. Increases in both lumbar spine and femoral neck BMD were found in patients treated with clodronate, although the only significant increase was observed in lumbar spine BMD at the end of 18 months. The mean changes in BMD were not significantly different, compared with the other groups, and at the end of 18 months there was a 2.20 +/- 0.58% increase at the lumbar spine. The mean vertebral and femoral neck BMD did not change significantly throughout the study period in patients receiving calcitonin. At the end of 18 months, there was a 0.13 +/- 0.52% decrease and a 0.11 +/- 0.49% increase in mean lumbar spine and femoral neck BMD, respectively, compared with baseline. The combination of clodronate plus calcitonin increased mean lumbar spine and femoral neck BMD by 2.08 +/- 1.05% and 1.46 +/- 1.09%, respectively, at the end of 18 months, but these increases were not significantly different from those in the groups where these agents were used alone. Significant decreases in bone resorption and in markers of bone formation were observed in all groups. CONCLUSION: HRT was found to be the most effective treatment regimen in postmenopausal patients with osteopenia, compared with clodronate, calcitonin and a clodronate plus calcitonin combination. Clodronate or calcitonin might be alternatives when HRT is contraindicated or refused by the patient; although calcitonin was found to be less effective. The clodronate plus calcitonin combination was not superior to either of these agents when used alone. PMID- 11910659 TI - Factors affecting sexual functioning of women in the mid-life years. AB - OBJECTIVE: To model the interaction of hormones, symptoms and psychosocial factors on women's sexuality during the menopausal transition. DESIGN AND METHODS: A prospective, observational study was carried out of a community-based sample of 438 Australian-born women aged 45-55 years at baseline. The study comprised six annual assessments in the women's own homes utilizing a core questionnaire, with rating scales for well-being and daily hassles, and a Personal Experiences Questionnaire as a measure of sexual functioning. Levels of follicle stimulating hormone (FSH), estradiol and inhibin were measured annually. Statistical analysis was performed by structural equation modelling. RESULTS: The retention rate was 90% (final sample size after exclusions, n = 354). The normal fit index for the global model obtained was 0.92. There is a significant direct effect of menopausal status on vaginal dryness/dyspareunia, and an indirect effect on sexual responsivity via a direct effect of menopausal status on symptoms, which then affect well-being. Menopausal status reflects hormonal status. Feelings for the partner and the partner's sexual problems have direct effects on different aspects of sexual functioning. Other social variables such as paid work, interpersonal stress, daily hassles and educational level affect sexual functioning indirectly via effects on symptoms and well-being. CONCLUSIONS: Psychosocial factors, symptoms and the menopausal transition affect women's sexual functioning during the mid-life years. PMID- 11910660 TI - Out-patient hysteroscopy in asymptomatic postmenopausal women. AB - OBJECTIVE: To determine the prevalence of uterine and endometrial abnormalities in normal postmenopausal women and assess the accuracy of subjective hysteroscopic appearances and endometrial histology following 12 weeks of hormonal treatment. DESIGN: A multicentered randomized double-blind placebo controlled trial in which the volunteers received conjugated equine estrogen 0.625 mg, selective estrogen receptor modulator (in one of two doses) or placebo. SETTING: Out-patient endoscopy unit in a large teaching hospital. METHODS: Out patient hysteroscopy and endometrial biopsy. RESULTS: Forty-eight women had a hysteroscopy and endometrial biopsy: eight (16.6%; 95% confidence interval, CI 6 27%) women had intrauterine polyps and 38 (79.2%) women had normal hysteroscopies. Thirty-five women had a repeat endometrial assessment with hysteroscopy and, for the detection of proliferative endometrium (prevalence 22.8%), a sensitivity of 87.5%, a specificity of 74%, a negative predictive value of 95% and a positive predictive value of 50% were observed. There was a good proportion of agreement, 0.77 (95% CI 0.63-0.91), but a kappa score of 0.486 revealed only a moderate level of agreement. The likelihood ratios for proliferative endometrium were: LHR+ = 3.38 (fair), and LHR- = 0.17 (moderate). CONCLUSIONS: Hysteroscopic assessment of the uterine cavity is efficient in the detection of pathological intrauterine lesions, but is only moderately successful in determining physiological changes in the endometrium. This study defines a standard of observational statistics for out-patient hysteroscopy in relation to normal endometrial histology in postmenopausal women who may have been ingesting exogenous estrogens. PMID- 11910661 TI - Selective estrogen receptor modulators: tissue selectivity and differential uterine effects. AB - Selective estrogen receptor modulators (SERMs) are compounds that bind to estrogen receptors and produce estrogen-like (agonist) effects in some tissues and estrogen-blocking (antagonist) effects in other tissues. One of the goals of SERM research has been to develop compounds that provide the potential benefits of estrogen in the skeleton and cardiovascular system, but avoid the negative effects of estrogen in other tissues. Estrogen therapy has been consistently associated with endometrial stimulation, including glandular proliferation, hyperplasia and cancer. In contrast, the presence or degree of endometrial stimulation observed with SERMs varies by compound. The purpose of this review is to differentiate the endometrial effects of compounds that display a SERM-like profile. Molecular mechanisms involving SERM binding to estrogen receptors, preclinical uterine effects in both tissue culture and animal models, and endometrial findings in clinical experience are discussed. There are several SERMs commercially available or in development. The favorable safety profile of raloxifene in the uterus differentiates it from the others. Future SERM development will continue to focus on finding compounds that exhibit minimal endometrial stimulation. PMID- 11910662 TI - Bleeding patterns and endometrial histology during administration of low-dose estradiol sequentially combined with dydrogesterone. AB - OBJECTIVE: To determine bleeding patterns and endometrial response in postmenopausal women taking low-dose (1 mg) estradiol in a sequential combined formulation with 5 or 10 mg dydrogesterone. METHODS: A total of 151 postmenopausal women were allocated randomly to 5 or 10 mg dydrogesterone during cycle days 15-28 in a sequential oral formulation with 1 mg estradiol continuously during 13 cycles of 28 days. Occurrence of vaginal bleeding was recorded daily and analyzed in accordance with World Health Organization (WHO) standards. Endometrial biopsies, obtained at baseline and cycle days 25-27 of the final treatment cycle, were interpreted independently by two pathologists. RESULTS: The study was completed by 131 women (87%). The percentage of women with bleeding (mean +/- SD) was 57.2 +/- 3.6% in the 1/5-mg group and 65.8 +/- 4.2% in the 1/10-mg group (p < 0.001); cross-sectional analysis showed that, in every cycle, there were more women with bleeding in the 1/10-than in the 1/5-mg group (p < 0.001). With regard to the day of onset of bleeding, the mean difference between groups was 1.4 +/- 1.1 days (p < 0.001). There was no difference in duration of bleed (5 days), or intensity or incidence of intermittent bleeding (3 14% per cycle). Both regimens resulted in high rates of amenorrhea in each cycle (26-49%), but only 14/151 (9%) women were amenorrheic throughout. Three patients (2%) discontinued owing to bleeding problems. Endometrial protection was adequate in 98.3% (1/5-mg group) and 98.5% (1/10-mg group) with only one case of proliferation (1/10-mg group) and one of simple hyperplasia (1/5-mg group). CONCLUSIONS: The bleeding pattern associated with low-dose (1 mg) estradiol sequentially combined with 5 or 10 mg dydrogesterone shows a high rate of amenorrhea in each cycle; there is a dydrogesterone dose effect on the occurrence and day of onset of bleeding. Bleeding episodes that occur show a regular pattern and are of slight intensity. The endometrial safety of both regimens is high. PMID- 11910663 TI - Patient acceptability of and tolerance to a placebo intravaginal ring in hysterectomized women: a pilot study. AB - OBJECTIVE: To assess the tolerance to and patient acceptability of a placebo intravaginal ring in hysterectomized, postmenopausal women over a 28-day period. DESIGN: Open, single-center. SETTING: University hospital specialist menopause clinic. PARTICIPANTS: A total of 24 healthy postmenopausal women attending menopause clinics at King's College Hospital and the Amarant Centre, London. MAIN OUTCOME MEASURES: Discomfort, ring expulsions, bowel, urinary and sexual difficulties associated with the ring and women's overall views on this method. RESULTS: The major problems with use of the intravaginal ring were patient discomfort, involuntary ring expulsion and sexual partner discomfort. These were experienced by 13, 11 and seven women and their partners, respectively, and resulted in premature discontinuation by three women who suffered discomfort and two women who experienced recurrent involuntary ring expulsion. CONCLUSIONS: Two thirds of the patients found the intravaginal ring acceptable, one-third did not. PMID- 11910664 TI - Serum estrogen level, attention, memory and other cognitive functions in middle aged women. AB - OBJECTIVE: To evaluate the relationship between serum estradiol level and cognitive processing efficiency and memory. METHODS: Sixty-three healthy women aged 45-65 years were recruited through a newspaper announcement. The subjects were divided into two subgroups (low-estrogen group, n = 37 and high-estrogen group, n = 26) according to their serum follicle stimulating hormone (FSH) and estradiol levels. In the high-estrogen group, estrogen was either endogenous or supplied by estrogen replacement therapy. Automatic and controlled cognitive processing and attentional resources were measured using CogniSpeed software, together with conventional tests of cognitive performance: similarities, digit span, digit symbol, block design, object naming and recall, paired word associates (PWA) recall, Benton visual retention and paced auditory serial addition test (PASAT). The Beck depression inventory was also assessed. RESULTS: Cognitive reaction speeds were similar in both groups. Women with low estrogen levels made more errors in the vigilance test (sustained attention, p = 0.040). There were no differences in short-term or long-term memory, or verbal, visual or working memory between the study groups. Older women were slower in the ten choice reaction time (10-CRT) test (r = 0.25, p = 0.047) and made more errors in the test of suppressing attention (Stroop incongruence test; r = 0.34, p = 0.007) and in the sustaining attention test (vigilance test; r = 0.47, p < 0.001). Depression scores did not correlate with cognitive variables. CONCLUSIONS: Cognitive performance was well preserved in healthy middle-aged women. Cognitive speed, accuracy, attentional resources and memory did not show impairment with decline of serum estrogen level in this age group. PMID- 11910665 TI - Changes in coagulation factors and fibrinolytic components of postmenopausal women receiving continuous hormone replacement therapy. AB - OBJECTIVE: The effect of hormone replacement therapy (HRT) on coagulation factors and fibrinolytic components in postmenopausal women was studied for 6 months to elucidate whether continuous HRT has an influence on thrombosis. METHODS: One hundred and thirty-four postmenopausal women were divided into three groups according to treatment: 39 women who had undergone hysterectomy and oophorectomy received 0.625 mg/day of conjugated equine estrogen (CEE) continuously (CEE therapy), 48 postmenopausal women received both 0.625 mg/day of CEE and 2.5 mg/day of medroxyprogesterone acetate (MPA) continuously (CEE/MPA therapy) and 47 postmenopausal women received placebo as control. The following variables were measured before treatment as well as after 1, 3 and 6 months of treatment: factor VII activity, protein C activity, fibrinogen level, antithrombin III activity, plasminogen activator inhibitor-1 (PAI-1) level and the plasma concentration of tissue-type plasminogen activator (t-PA). RESULTS: After 1 month of treatment, protein C activity increased by 9.6% and 11.4% of the initial value (p < 0.05), fibrinogen level decreased by 7.8% and 6.1% of the initial value (p < 0.05) and PAI-1 decreased by 19.4% and 14.3% of the initial value (p < 0.05) in the CEE therapy group and the CEE/MPA therapy group, respectively. Factor VII activity increased by 10.1% of the initial value (p < 0.05) in the CEE therapy group only. Antithrombin III and t-PA levels did not change throughout either treatment. CONCLUSION: Except for an increase in factor VII activity in the case of continuous CEE therapy, continuous HRT had no unfavorable effects on either coagulation factors or fibrinolytic components. PMID- 11910666 TI - Life satisfaction and health-related quality of life (SF-36) of middle-aged men and women. AB - OBJECTIVES: To investigate life satisfaction and health-related quality of life (SF-36) in a general population sample of middle-aged women and men. The effects of menopausal status and hormone replacement therapy (HRT) use upon life satisfaction and health-related quality of life (HrQOL) were also examined for the female sample. DESIGN: All men and women aged 49-55 years from the age/sex register of a large general practice in London were contacted and asked to complete a questionnaire about their health. SUBJECTS: A total of 103 women (55%) and 86 men (40%) participated; of the women, 15% were premenopausal, 68% peri- or postmenopausal and 17% taking HRT. RESULTS: Women and men reported similar levels of HrQOL, life satisfaction and general health, although women reported more physical problems (SF-36). The significant predictors of HrQOL were serious illness, employment and marital status, but HRT use and menopausal status were not significantly associated with life satisfaction nor HrQOL (for women). CONCLUSIONS: Gender differences in health and HrQOL may be less apparent during mid-life, although there were some subtle differences between men and women in reported health concerns and reasons given for (dis)satisfaction with their lives. PMID- 11910667 TI - Polycystic ovary syndrome and implications for the menopause. AB - Polycystic ovary syndrome (PCOS) is the most common endocrine condition of premenopausal women and has significant metabolic abnormalities that could have an impact after the menopause. Diabetes mellitus, hyperlipidemia and hyperinsulinemia could potentially affect health in this era of life. Endometrial cancer, due to unopposed estrogen action, is more common where progestins have not been given for menstrual dysfunction. Preventive management earlier in life will avoid postmenopausal problems in PCOS. PMID- 11910668 TI - Menopausal knowledge and perception in the Bulgarian population. PMID- 11910669 TI - The four harms of harmless therapies. PMID- 11910670 TI - Is there a proven place for phytoestrogens in the menopause? PMID- 11910671 TI - The effect of Promensil, an isoflavone extract, on menopausal symptoms. AB - OBJECTIVES: The primary aim was to assess whether the use of an isoflavone extract containing 40 mg or 160 mg of total isoflavones affects the frequency of menopausal flushes and other symptoms. The secondary aims were assessments of possible effects on menopause symptom scores and biological measures of estrogen activity. METHODS: A randomized, double-blind, placebo-controlled prospective trial of 37 postmenopausal women with symptoms of estrogen deficiency was performed over a 12-week period. The women were randomized to three treatment groups: placebo, 40 mg or 160 mg, delivered in tablet form. RESULTS: There was no significant difference in the incidence of flushes between the three groups at trial conclusion. There was no difference between the groups in Greene Menopause Symptom Scores, vaginal pH, levels of follicle stimulating hormone (FSH), sex hormone binding globulin (SHBG) or total cholesterol, liver function or blood parameters. A statistically significant increase in high-density lipoprotein (HDL) cholesterol of 18.1% (p = 0.038) occurred in the 40-mg group. CONCLUSION: A large placebo response and inadvertent use of dietary isoflavones in the placebo group may have obscured a significant change in flushing frequency. Previous uncontrolled studies claiming a beneficial effect of foods with a high isoflavone content on menopausal symptoms may have been confounded by a large placebo response. PMID- 11910672 TI - Randomized placebo-controlled trial of an isoflavone supplement and menopausal symptoms in women. AB - OBJECTIVE: To test the hypothesis that increasing the intake of isoflavones by dietary supplementation may produce a therapeutic effect in reducing the incidence and severity of hot flushes in menopausal women. METHODS: Fifty-one postmenopausal women were randomized to placebo and active (one tablet per day of a 40-mg isoflavone supplement) groups in a cross-over design trial. After a 1 week run-in period, subjects were commenced on a 12-week period of treatment (active or placebo), followed by a 1-month placebo wash-out period, then crossed over to the alternative treatment regimen for a further 14 weeks. Symptom diaries were maintained throughout the trial and at the start and end of treatment. Plasma sex hormone binding globulin (SHBG) assay, full blood count, biochemical profiles, vaginal swabs and vaginal ultrasound scans were performed and isoflavones determined in 24-h urine collections by high-pressure liquid chromatography (HPLC) analysis. RESULTS: There was no significant difference between active and placebo groups in the reduction in hot flushes between start and finish time-points. Analysis performed on interim data time-points revealed a substantially greater reduction in flushing in the active group than placebo at 4 and 8 weeks after commencement of treatment, but this was not statistically significant. There were no significant differences between groups for Greene scores or in SHBG levels, hematological or biochemical parameters and vaginal swab or ultrasound findings. The combined values for all subjects, regardless of treatment group, revealed a strong negative correlation between the level of urinary isoflavone excretion and the incidence of hot flushes. CONCLUSIONS: These data do not indicate a therapeutic benefit from dietary supplementation with isoflavones in women experiencing menopausal symptoms, but do indicate that the apparent placebo effect in many studies of menopausal symptoms may be attributable to dietary sources of isoflavones. The study also demonstrates that 3 months of isoflavone supplementation did not cause adverse events or endometrial changes. PMID- 11910673 TI - Long-term effects of oral estradiol and dydrogesterone on carbohydrate metabolism in postmenopausal women. AB - OBJECTIVE: To determine in postmenopausal women the long-term effects on carbohydrate metabolism of the administration of oral micronized 17 beta estradiol (2 mg/day continuously) and cyclical dydrogesterone (10 mg/day for 14 days per 28-day cycle). METHODS: A 2-year open-label prospective, non-comparative study was carried out of 13 healthy postmenopausal women receiving cyclical estradiol and dydrogesterone and serving as their own controls. Concentrations of blood glucose, plasma insulin, C-peptide, glucagon and free fatty acids (FFAs) were determined before treatment (base-line) and at 6, 12 and 24 months of hormone replacement therapy under fasting conditions and during a standard 75-g, 3-h, oral glucose tolerance test (OGTT). RESULTS: Fasting blood glucose levels were unchanged throughout the study, and the mean areas under the curves (AUCs) for glucose response increased slightly but non-significantly versus baseline; fasting plasma insulin levels tended a decrease, and AUCs for insulin responses to the glucose load fell by 23% from baseline (not significant); fasting C peptide levels and AUCs were unchanged; plasma glucagon fasting levels and responses were in the normal range and stable throughout the study; and plasma FFA fasting levels decreased significantly, as well as FFA AUCs during OGTTs, at the 12th and 24th months of the study. CONCLUSIONS: During a 2-year treatment with oral estradiol and cyclical dydrogesterone, a direct progesterone derivative, tolerance to glucose was unchanged, fasting plasma insulin and insulin response to repeated glucose loads were decreased, and C-peptide levels remained unchanged, indicating a potential improvement in insulin sensitivity and clearance, as in younger women; additionally, a slightly enhanced antilipolytic activity of insulin was observed. PMID- 11910674 TI - High resolution ultrasound assessment of the carotid artery: its relevance in postmenopausal women and the effects of tibolone on carotid artery ultrastructure. AB - OBJECTIVES: Hormone replacement therapy protects from cardiovascular disease at the menopause in part by reduction of menopausal pro-atherogenic serum lipid changes. Tibolone has beneficial effects on lipids, although serum high density lipoprotein levels decrease. This study aimed primarily to establish the effects of long-term administration of tibolone on a new surrogate marker for cardiovascular disease risk, the measurement of carotid artery intima-media thickness (CIMT) using high-resolution ultrasound. METHODS: Measurement of CIMT and assessment of carotid atherosclerotic plaques were undertaken in 31 women on tibolone and 30 voluntary controls from an ongoing open-label study of tibolone. RESULTS: The two groups were comparable, except for mean age and prevalence of current smokers. Repeatability of CIMT measurements was acceptable (CV, 10.0%). CIMT was significantly thicker in those with atherosclerotic plaques and increased systolic blood pressure. Prevalence of plaques was raised in those who had ever smoked, and those with elevated systolic blood pressure. There was no influence of tibolone on CIMT, whether plaques were present or not. CONCLUSIONS: This reliable technique demonstrates associations between CIMT and established risk factors. CIMT was significantly thicker in those with existing plaques. We did not demonstrate an effect of long-term tibolone use on either CIMT or prevalence of plaques. PMID- 11910675 TI - HERS and its aftermath. PMID- 11910676 TI - Dispositional factors, coping and adaptation during menopause. AB - The objective of the study was to examine the direct and indirect influences of dispositional factors, namely optimism, health-related hardiness (HRH) and sense of coherence (SOC), on the symptom experiences of peri- and postmenopausal women. Indirect effects of dispositional factors were examined via attitudes to the menopause and coping (emotion-versus problem-focused). A survey methodology was employed to assess the research objective for 176 peri- and postmenopausal women recruited from menopause clinics and family planning centers in Queensland, Australia. Emotional stability of the subjects was statistically controlled to eliminate any possible confounding effect on symptom reporting. The results indicated that optimism and SOC affect menopausal health directly, as evidenced by fewer symptoms reported by women scoring highly on these dispositions. Any indirect effect of HRH, optimism and SOC appears to be exerted via problem focused coping rather than emotion-focused coping or through attitudes. It was concluded that dispositional factors are important to the experience of the menopause and how women adapt to their midlife transition. Psychologists and professionals working in menopause clinics may need to promote feelings of optimism and a sense of coherence in menopausal women, to facilitate better adaptation to this important transitional phase in women's lives. PMID- 11910677 TI - Climacteric modifications in body weight and fat tissue distribution. AB - OBJECTIVE: To estimate the effects of climacteric modifications on body weight and fat distribution. METHODS: From 8764 women attending the authors' Menopause Clinic, 1075 untreated, normal healthy women were selected and divided into three groups: premenopausal (n = 380), perimenopausal (n = 263) and postmenopausal (n = 432). The total body fat tissue mass and distribution were analyzed using the dual energy X-ray method. RESULTS: Body weight and body mass index (BMI) were significantly higher in perimenopausal and postmenopausal than in premenopausal women. The mean total body fat and the percentage of fat with respect to soft tissue were significantly (p < 0.05) higher in the perimenopausal and postmenopausal groups than in the premenopausal group. The amount of fat tissue and the regional fat percentage with respect to total fat tissue were higher in the trunk (p < 0.05) and arm (p < 0.05) regions in perimenopausal and postmenopausal than in premenopausal women. In the postmenopausal group, the leg fat tissue was significantly (p < 0.05) less than in the premenopausal and perimenopausal groups. Total body and leg lean tissue was significantly (p < 0.01) less in postmenopausal than in premenopausal and perimenopausal women. In the premenopausal group, body weight and BMI were positively correlated with age (r = 0.37 and r = 0.54, respectively). No significant correlations were observed in the perimenopausal group. In postmenopausal women, body weight and BMI were loosely correlated with age (r = 0.13 and r = 0.11, respectively). In three groups of 63 age-matched women, with similar BMI, the percentage of total body fat with respect to soft tissue was significantly (p < 0.001) higher in the perimenopausal and postmenopausal groups than in the premenopausal group. Regarding body fat distribution, the percentage of fat with respect to total fat tissue was significantly higher in the trunk (p < 0.001) region in perimenopausal and postmenopausal women than in premenopausal women. In the leg region, the percentage of fat with respect to total fat tissue was significantly (p < 0.05) higher in the premenopausal group than in the postmenopausal group. In the arm region, a slight but not significant (p < 0.08) difference was shown in fat distribution among the three groups. CONCLUSIONS: Climacteric changes rather than the aging process are relevant for prediction of body weight and fat distribution, especially for perimenopausal and postmenopausal women, who show a shift to a central, android fat distribution. PMID- 11910678 TI - Changes in bone mineral density with age, menopausal status and body mass index in Turkish women. AB - OBJECTIVE: The aim was to evaluate changes in bone mineral density (BMD) with age and body mass index (BMI) in healthy pre- and postmenopausal women living in the urban areas of Turkey. DESIGN: The study was prospective, carried out from 1993 to 1997. SETTING: The study carried out at a university hospital, the Istanbul University Cerrahpasa School of Medicine, Turkey. PATIENTS: The study group consisted of 849 healthy women of ages 20-84 years, admitted to the Istanbul University Cerrahpasa School of Medicine. The cases were divided into age groups, starting with 20-29 years and ending with 70 years and over. For regression analysis, the cases were further regrouped as 20-39, 40-59 and 60 years and over. Dual energy X-ray absorptiometry (DEXA) was used to measure BMD in the lumbar vertebrae (L2-L4) and in the classical locations of the proximal femur such as the femoral neck, the Ward's triangle and the trochanter. Multiple regression analysis was performed for the evaluation of annual changes in BMD with respect to age and/or BMI. RESULTS: A significant decrease in BMD started especially in the 40-49 age group, matching the average age of menopause in the study population. In contrast to the non-significant changes in the 20-39 age group, a significant decrease in BMD in the 40-59 age group, which included the average age of menopause, was detected in all locations (p < 0.0001). In addition to the significant effect of the menopause on BMD, an association between BMD and BMI was found in every location and age group (p = 0.02 to p < 0.0001). The total bone loss in the 70 and over age group, in comparison with the 30-39 age group, was 18.78% in L2-L4, 21.69% in the femoral neck, 32.68% in the Ward's triangle and 14.11% in the trochanter. Corresponding values between age groups 70 and over and 60-69 were 0.25%, 7.62%, 11.94% and 8.29%, respectively. Women in the older age groups had a slower decline in BMD in the lumbar vertebrae, in comparison with the proximal femur. Moreover, the maximum postmenopausal total bone mineral loss was in the Ward's triangle. CONCLUSIONS: The present results, confirming the results of other studies, have revealed a significant association between BMD and the menopausal status of women in the Turkish population. Additionally, a significant correlation has been detected between BMI and BMD, regardless of location and age. PMID- 11910680 TI - The HERS trial. PMID- 11910679 TI - Regulation of endometrial angiogenesis. AB - Bleeding problems are the most common reason for discontinuation of hormone replacement therapy. Human endometrium undergoes the unique process of benign angiogenesis under the control of ovarian steroids during reproductive life and it is presumed that similar processes occur when women take hormone replacement therapy. The key players in endometrial growth and angiogenesis are vascular endothelial growth factor, thymidine phosphorylase and adrenomedullin. Regulation of these angiogenic factors is described. PMID- 11910681 TI - Hot flushes and other menopausal symptoms in relation to soy product intake in Japanese women. AB - OBJECTIVE: To examine the relationships between dietary intake of soy products and hot flushes and other menopausal symptoms. METHODS: Subjects were 284 women aged 40-59 years who attended a health check-up program provided by a general hospital in Gifu, Japan. They completed a health questionnaire including the Kupperman test of menopausal distress. Diet was assessed by a semiquantitative food frequency questionnaire. RESULTS: Fermented soy product intake but not total soy product intake was significantly negatively correlated with hot flush severity (r = -0.16, p = 0.01) after controlling for age and menopausal status. Neither total soy product intake nor fermented soy product intake was significantly correlated with menopausal index score. Estimated isoflavone intake from total and fermented soy products was significantly lower by 15% (p = 0.02) and 19% (p = 0.01), respectively, in women with hot flushes, compared to those without hot flushes after controlling for covariates. CONCLUSION: The data support a hypothesis that intake of fermented soy products alleviates the severity of hot flushes. PMID- 11910682 TI - Some dental history factoids.... PMID- 11910683 TI - Managed care in dentistry brings antitrust issues to spotlight. PMID- 11910684 TI - Changes in dentistry. PMID- 11910685 TI - [Bronchial caliber study: a continuous challenge in pneumology]. AB - The caliber and the bronchial wall represent the anatomical substratum of several diseases of the respiratory tract. Many investigations allow qualitative and quantitative approaches but none can appreciate exactly the injuries degree and the lesions extent. The radiological methods are powerful in particular the high resolution computed tomography with great in-depth descriptive capacity. The respiratory functional tests with a variable sensitivity allow a dynamic but indirect analysis, the provocation test studies bronchial hyperresponsiveness and the plethysmography measures the airways specific resistance. The bronchoscopy explores the bronchi directly but is confronted with the problem of the image distortion. The fields of studies of the bronchial gauge are vast including all bronchopulmonary pathology and the correlation of two different methods allows a better evaluation. PMID- 11910686 TI - ["Reforms of the Tunisian health system and health insurance"]. AB - This article introduces on-going reforms and the perspectives of evolution of the tunisian health system. After a review of how Tunisia is redefining its health system in two directions: management system and funding procedures, the authors focussed on the analysis of perspectives of health insurance and a unified health coverage system. The solution of health system problems cannot be found before undertaking a precise reformulation of health system objectives, such a reformulation needs to imply all contributing persons and institutions in order to preserve accessibility, equity, quality and performance. In fact, it is a requested to split from the current diverse and pluralistic system to unified and homogenous new system. PMID- 11910687 TI - [Basis and guidelines for empirical antibody therapy in the management of community-acquired pneumonia]. AB - The choice of the antibiotic should be based on clinical, chest X-ray radiography and essentially microbiologic criteria. Incurrent practice treatment is more often empiric based on epidemiologic characteristics of the microbiologic agents and the particularities of each patient. A satisfactory approach requires, in addition, a perfect knowledge of different available antibiotics and the resistance of certain etiologic pathogens to these latters. because S. Pneumoniae is the most frequently encountered pathogen, B lactams and especially Penicillin G. and amoxicillin remain the most useful drugs prescribed for adults with risk factors. However, in advanced age patients and those with comorbidity, the spectrum should be enlarged and should include, besides S. Pneumoniae, H. influenzae and other Gram negative bacilli. When the pneumonia is more severe and has required hospitalization, the antimicrobial therapy must be immediate, multiple and large. The causal agent must be searched for desperately, so that the antimicrobial therapy can be adapted secondary to the results of the antibiogram. PMID- 11910688 TI - [Allogenic hematopoietic stem cell transplantation in acquired aplastic anemia: first experience of the National Center for Bone Marrow Grafting]. AB - Bone marrow transplantation from HLA-identical sibling offers cure and leads to restoration of normal hematopoiesis and long-term survival in 60-80% of recipients. From february 1998 to october 1999, seven patients with aplastic anemia (2 very severe aplastic anemia and 5 severe aplastic anemia), with a median age of 22 years (14-39), received a transplant from an HLA-identical sibling donor. All patients had sustained engraftment. Only one patient developed grade IV acute graft-versus-host disease. One patient died in the 22th day of systemic mycobacterial infection and one in the 79th day of acute graft-versus host disease. The remaining 5 patients are alive and have a complete hematological recovery, with a median follow-up of 6 months (1,5-12). There are at least two reasons for the improved survival of patients with aplastic anemia who where treated by HLA-indentical bone marrow transplantation. One is the decreased incidence of graft rejection that has resulted from the more judicious use of transfusions before bone marrow transplantation, and improvements in the immunosuppressive qualities of the conditioning programs. Another reason for improved survival is the decrease in the incidence and severity of acute graft versus-host disease. PMID- 11910689 TI - [Control of hemodynamic variations secondary to suspension microlaryngoscopy]. AB - The objective was to evaluate the clinic interest of the esmolol in the attenuation of the hemodynamic response to suspension microlaryngoscopy (mls). It was a prospective, randomized and double blind study, having included 29 patient ASA II and III, distributed in two groups, having received respectively before laryngoscopy: group A (n = 15), esmolol in intravenous (i.v.) at the dose of 150 micrograms.kg-1 and DS group (n = 14), 10 ml of normal saline at 0.9%. Has been achieved a general anaesthesia i.v. based on propofol, fentanyl and atracurium. After tracheal intubation, it has been assured an artificial ventilation. The two groups were comparable for the demographic and anesthetic characteristics. A significant decrease of the mean arterial pressure (-11%) and of the prp (product systolic arterial pressure heart rate) have been raised during the mls, in the group esmolol, whereas the variations of the cardiac frequency were comparable. In order to assure a better hemodynamic control during the mls, it seems important, in addition to the adjustment of the recommended dose of esmolol, to cover the whole duration of the endoscopic act by a continuous drip. PMID- 11910690 TI - [Ramadan and customs of life: investigation with 84 adult residents in the district of Tunis]. AB - To know nature and the dimension of the change of the customs of life leads by the Ramadan, we led a comparative descriptive inquiry before and during the month of the fast at 84 adults residents in the district of Tunis. Our results underline an increase of the consumption of meat and eggs with an average frequency of 4.3 and 6.1 times a week respectively. This overconsumption of the animal proteins contrasts with a tendency in the decline of the consumptions of vegetables. The exciting (tea, coffee, tobacco) are less consumed during the Ramadan. Also, we noted a decline of 50% of the average number of smoked cigarettes. There is an intensification of domestic links with an increase of the frequency of exchange of domestic visits. It's crossed of average from 0.7 to 1.2 times a week (p < 0.001). The phenomenon of irritability is frequently lived by near 20% of the investigated. We recommend under shape of an educational program the intensification of positive customs and the correction of negative customs, to benefit of sacred month, to tighten towards a more balanced life all year. PMID- 11910691 TI - [Management of uterine fibromas. Report of 219 cases]. AB - This study is about 219 uterine fibroma treated surgically between 1994 and 1998. The surgical operation represent 19.1% of the gynecological interventions. The average age of our patients was of 41.7 years with 30.6% of nulliparous. The main motives of consultation were: the confusion of the menstrual cycle in 56.6%, the pelvic pains in 32%, the increase of the volume of the belly in 10% and the infertility in 10% of cases. The surgical indications were dominated by the big size of the womb in 68% of cases and hemorrhagic complications in 57.7% of cases. The myomectomy indicated for women young and avid for pregnancy was realized in 94 cases; 80.8% by abdominal way, 13.8% by hysteroscopic way and 5.3% by coelioscopy. Hysterectomy proposed for women near menopauses was realized in 125 cases: 70.4% by abdominal way and 29.6% by vaginal way. The complications per- and post-operating are rare, represented essentially by the bleeding and the urinary infections. PMID- 11910692 TI - Acute appendicitis during pregnancy. AB - Acute appendicitis is the most common surgical emergency occurring during pregnancy. To determine possible methods for improving diagnostic and management accuracy, a retrospective review was conducted of 23 pregnant patients who underwent laparotomy with a preoperative diagnosis of acute appendicitis. Gestational stage at presentation included the first trimester in 2 patients, the second trimester in 6 patients and the third trimester in 15 patients. Eighteen patients (78.26%) had pathologically proven acute appendicitis. Perforation occurred in 2 patients. Postoperative fetal complications included one intrauterine death (4.3%) and three premature births. There was no maternal deaths and morbidity was limited to atelectasis and wound infection in 4 patients. Gestational physiologic changes make difficult the diagnosis of acute appendicitis. Prompt surgical intervention prevent maternal complications and foetal loss. PMID- 11910693 TI - [Diagnosis of holoprosencephalia. Report of 17 cases]. AB - OBJECTIVE: To establish the epidemiologic profile of holoprosencephalia and determine benefits of ultrasound and foetopathologic examination to the diagnostic. METHODS AN MATERIAL: [corrected] Retrospective study about 17 cases of holoprosencephalia observed in CMNT between Janaury 1992 and September 2000. RESULTS: Ultrasound diagnosis was made in 13 cases (75%). Ultrasound criteria were; absence of median structure of the brain and unique ventricule. The prognosis was always bad. Foetopathologic examination revealed 7 cases of lobar holoproencephalia and 10 of semi lobar. Fascial dysmorphia were noted in 82% of cases. CONCLUSION: The foetopathology and genetic counselling looking for fascial, dysmorphia in family's members gives a good evaluation of recurrences. PMID- 11910694 TI - [Pregnancy and AIDS. Report of 9 cases]. AB - The pregnancy at women infected by the HIV characterizes by the risk of transmission of the HIV to the child. The goal of this retrospective study concerning 9 deliveries of women infected by the HIV cured in the Infectious Diseases Service and the Lalla Meryem's Maternity of Casablanca CHU Ibn-rochd during 10 years (1990-1999) is to specify the experience of two services of concerning plug in cost of the patients infected by HIV by trying to pull some a protocol of plug in cost. The antiretroviral therapy has been used in 8 cases out 9. The AZT long protocol has been used in 6 cases and the bitherapy in 2 cases. The delivery has taken place by the natural way under cover of AZT in all cases. All babies have received the AZT in syrup and none has been breastfed to the breast. Two newborns on 9 out 9 have been contaminated by the HIV. PMID- 11910696 TI - [Brucella endocarditis of native valves. Report of 3 cases]. AB - Brucella endocarditis is a rare but a serious complication of human brucellosis. We report 3 cases, the diagnostic was suspected by the patient's history of systemic brucellosis in two cases and established by the culture of native valve material in the third. All the patients underwent surgery for non control of the infections, one patient died in immediately postoperative period by acute cardiac failure. For the other patients, there were no early or late mortality and no recurrence after a follow up of respectively 6 and 84 months. The diagnostic of brucella endocarditis needed a very high degree of clinical suspicion, it requires an early management valve replacement is in the majority of cases, followed by adequate and prolonged antibiotic treatment. PMID- 11910695 TI - [Fatty liver disease and other complications of obesity]. AB - Obesity induces many organic complications (somatic, metabolic and mechanical). The aims of this prospective study were to evaluate its complications in a group of thirty fat patients. Fatty liver disease, detected by echography is noted at 70% of patients. Clinically, it makes pains at the right side and seems favorized by an elevated BMI > 40 kg/m2, and diabetes, hypertriglyceridemiae, hypercaloric and hyperglucidic ration. 10% of our patients present stone bile duct associated to fatty liver disease. PMID- 11910697 TI - [In situ hybridization in undifferentiated nasopharynx carcinoma in Tunisia. Report of 3 cases]. AB - The undifferentiated carcinoma of nasopharyngeal type (UCNT) is highly prevalent in South East Asia countries and has intermediate incidence in Tunisia, where Epstein-Barr virus constitutes one of the key factors of oncogenesis. Our preliminary results, in 3 patients with UCNT, show an elevated rate of EBV antibodies and a latent infection (type II) (expression of LMP and absence of ZEBRA by immunohistochemical reaction) and a positive staining with EBERs probe by in situ hybridation (ISH) in all cases. These results confirm a close association between EBV and tunisian UCNT. Moreover, the use of HIS technique for detection of EBERs constitutes an additional and formal argument of this association. PMID- 11910698 TI - [Esophageal cancer with esophagotracheal fistula. Treatment by esophageal prosthesis. Report of two cases]. AB - Esophageal cancer has a bad prognosis because of late diagnosis. The rate of Survival is very low. Palliative treatment is frequently made. The aim of the treatment is to allow feeding, and treat some complications likes breathing troubles. We report two cases of esotracheal fistulae secondary to esophageal cancer. The esophageal fistula was successfully treated by esotracheal prosthesis. PMID- 11910699 TI - [Erosive spondylarthropathy in hemodialysis. A case report]. AB - We report a case of 67-year-old woman with 11-year history of hemodialysis that complain of neck pain associated with cervico-brachial neuralgia. Imaging finding simulate infectious spondylitis. We consider the diagnosis of destructive spondylarthropathy in hemodialyzed patient since etiologic investigation was negative and RMN finding was suggestive. Our patient was partially improved with symptomatic treatment. PMID- 11910700 TI - [Muscle metastasis of a primary bronchial carcinoma]. AB - Intra muscular metastasis are rare. They usually occurs during the course of a cancer. The primary bronchial carcinoma is the most common. Clinical features include painful mass, frequently, they are symptomatic. Sonography CT scan and MR imaging shows a non specific abnormally, but they defined their seat and their extension. The authors report a case of metastases of epidermoid bronchial carcinoma to left adductor muscle and gives a review of the literature. PMID- 11910701 TI - Alcohol and hepatitis C. AB - Infection with the hepatitis C virus (HCV) has become a leading cause of scarring of the liver (i.e., fibrosis) and cirrhosis in the United States. HCV-related cirrhosis (with its associated complications, such as liver cancer) is a major cause of death, although it develops slowly and occurs only in approximately one third of HCV-infected patients. Alcohol can exacerbate HCV infection and the associated liver damage by causing oxidative stress and promoting fibrosis, thereby accelerating disease progression to cirrhosis. Furthermore, alcohol may exacerbate the side-effects associated with current antiviral treatment of HCV infection and impair the body's immune defense against the virus. Of the HCV infected people who do not consume alcohol, only a minority progresses to severe liver disease and requires antiviral treatment. Because alcohol potentiates the fibrosis- and cancer-inducing actions of HCV, alcoholics are particularly vulnerable to HCV infection and most in need of treatment. PMID- 11910702 TI - Alcohol's effects on the risk for coronary heart disease. AB - Several studies have indicated that moderate drinkers have a lower risk of both nonfatal myocardial infarction and fatal heart disease than do abstainers. To determine whether alcohol truly prevents coronary heart disease or whether other factors may contribute to this observed relationship, researchers conducted a systematic literature review and a combined analysis (i.e., meta-analysis) of 42 published studies. This analysis found that consumption of up to two drinks per day can promote changes in the levels of molecules that reduce the risk of heart disease while also increasing the levels of certain molecules that promote heart disease. Alcohol also may affect the risk of heart disease by acting on other various other molecules involved in a variety of physiological processes related to heart disease. Finally, the relationship between alcohol consumption and heart disease may be modulated by genetic factors. PMID- 11910703 TI - Alcohol consumption and the risk of cancer: a meta-analysis. AB - Alcohol consumption has been linked to an increased risk for various types of cancer. A combined analysis of more than 200 studies assessing the link between alcohol and various types of cancer (i.e., a meta-analysis) sought to investigate this association in more detail. This meta-analysis found that alcohol most strongly increased the risks for cancers of the oral cavity, pharynx, esophagus, and larynx. Statistically significant increases in risk also existed for cancers of the stomach, colon, rectum, liver, female breast, and ovaries. Several mechanisms have been postulated through which alcohol may contribute to an increased risk of cancer. Concurrent tobacco use, which is common among drinkers, enhances alcohol's effects on the risk for cancers of the upper digestive and respiratory tract. The analysis did not identify a threshold level of alcohol consumption below which no increased risk for cancer was evident. PMID- 11910704 TI - Alcohol and female puberty: the role of intraovarian systems. AB - Alcohol consumption during early adolescence may suppress the secretion of specific female reproductive hormones, thereby delaying puberty and adversely affecting the maturation of the reproductive system. These effects occur through several mechanisms, including altered production and secretion by the ovaries of estradiol, a key steroid hormone involved in the timing and regulation of female reproductive events. Alcohol can affect estradiol production by interfering with the normal function of regulatory hormones produced by the brain and the pituitary gland. Recent research has demonstrated additional potential mechanisms for alcohol's effects on female reproductive capability, including interference with specific regulatory systems located entirely within the ovary. Such "intraovarian" systems include the insulin-like growth factor-1 (IGF-1) and nitric oxide (NO) systems. Alcohol can dampen the stimulatory effects of the ovarian IGF-1 system and can increase the inhibitory effects of the ovarian NO system. These effects combine to decrease estradiol secretion. Thus, alcohol impairs ovarian function not only by interfering with hormonal communication between the brain, pituitary gland, and ovaries but also by directly altering the function of regulatory systems within the ovaries themselves. These results provide further evidence of the risks of underage drinking and the importance of its prevention. PMID- 11910706 TI - Alcohol and the male reproductive system. AB - Alcohol use affects all three parts of the hypothalamic-pituitary-gonadal (HPG) axis, a system of endocrine glands and hormones involved in male reproduction. Alcohol use is associated with low testosterone and altered levels of additional reproductive hormones. Researchers are investigating several potential mechanisms for alcohol's damage. These mechanisms are related to alcohol metabolism, alcohol related cell damage, and other hormonal reactions associated with alcohol consumption. Chronic alcohol use in male rats also has been shown to affect their reproductive ability and the health of their offspring. PMID- 11910705 TI - Effects of alcohol use and estrogen on bone. AB - In marked contrast with men who drink, women who drink alcohol are found, as a group, to have higher bone mass compared with women who abstain. Furthermore, the apparent beneficial effects of alcohol use are more apparent in postmenopausal women than women of reproductive age, suggesting that there might be an interaction between alcohol and estrogen. Estrogen deficiency accompanying menopause leads to bone loss, which in turn predisposes women to osteoporosis later in life. Estrogen deficiency accelerates bone remodeling, which is the process by which small areas of bone are destroyed and rebuilt, and leads to an imbalance whereby bone resorption--the part of remodeling consisting of breaking down and assimilating--exceeds bone formation. Alcohol might reduce bone loss in postmenopausal women by increasing the circulating levels of estrogen. Alternatively, alcohol might slow bone loss by acting on bone cells to reduce bone remodeling. Alcohol use has a negative effect on the immature skeleton but current understanding suggests that small quantities of alcohol may have beneficial effects on bone in older women. PMID- 11910707 TI - Effects of alcohol and HIV infection on the central nervous system. AB - Many people at risk for or infected with the human immunodeficiency virus (HIV) are heavy drinkers. Both HIV infection and heavy alcohol use adversely affect the immune system and central nervous system (CNS) function. However, little research has addressed the effects of heavy alcohol use on the severity and progression of HIV disease, including the development of HIV-associated CNS disease. Animal and in-vitro studies suggest that alcohol impairs various aspects of the immune system and increases the susceptibility to HIV infection, but may not accelerate progression of HIV disease. However, heavy alcohol use may interfere with the patient's adherence to antiretroviral treatment regimens. Neuropathological and neuropsychological studies have indicated that certain brain areas are affected by both HIV-infection and chronic alcohol abuse. Magnetic resonance spectroscopy studies of both HIV-positive and HIV-negative people who were either heavy or light drinkers found that chronic alcohol abuse exacerbates some metabolic injury in the brains of HIV-infected people, although this effect may be less pronounced in patients receiving effective antiretroviral therapy. PMID- 11910708 TI - Alcohol use and the risk of developing Alzheimer's disease. AB - Some of the detrimental effects of heavy alcohol use on brain function are similar to those observed with Alzheimer's disease (AD). Although alcohol use may be a risk factor for AD, it is difficult to study this relationship because of similarities between alcoholic dementia and AD and because standard diagnostic criteria for alcoholic dementia have not yet been developed. Similar biological mechanisms may be involved in the effects of AD and alcohol abuse on the brain. Epidemiologic studies have investigated the relationship between alcohol use and AD but have not provided strong evidence to suggest that alcohol use influences the risk of developing AD. Further research is needed before the effect of alcohol use on AD is understood fully. PMID- 11910710 TI - Pt. sexually assaulted by physician within 2-3 feet of nurses. PMID- 11910709 TI - Were nurses fired for refusing to participate in fraud? PMID- 11910711 TI - MA: 'school nurses' included in bargaining unit: court rules arbitrator exceeded authority. PMID- 11910713 TI - State regulations on agency nurse rates invalid. PMID- 11910712 TI - NC: did 'qualified immunity' apply to nurses?: court finds no 'deliberate indifference'. PMID- 11910714 TI - Economic analysis of a pragmatic randomised trial of home visits by a nurse to elderly people with hypertension in Mexico. AB - OBJECTIVE: To analyse the costs and the effectiveness of an intervention of home visits made by nurses to elderly people versus usual care given by the family medicine units. MATERIAL AND METHODS: A sample of 4,777 subjects aged 60 years and over covered by the Mexican Institute of Social Security (Instituto Mexicano del Seguro Social, IMSS) were screened. Those with a systolic and/or diastolic blood pressure level higher or equal than 160/90 mm Hg were randomly allocated to the intervention or control groups. The intervention consisted of visits at home by nurses who gave health and lifestyle advice to the participants. The economic evaluation was considered from a health services and patient perspective. Direct and indirect costs were calculated as incremental. Effectiveness was measured in terms of cost per millimetre of mercury reduced. RESULTS: Three hundred and forty five participants were allocated to the intervention group and compared with 338 controls. At the end of the intervention period the difference in the mean change in systolic blood pressure was 3.31 mm Hg (95% CI 6.32, 0.29; p = 0.03) comparing with the control group. In diastolic blood pressure the difference was 3.67 (95% CI 5.22, 2.12; p < 0.001). The total cost of the intervention was 101 901.66 pesos. The intervention cost per patient was 34.61 pesos (US$3.78), (CI 95% 34.44, 35.46). The cost-effectiveness ratios was 10.46 pesos (US$1.14) for systolic (CI 95% 129.31, 5.51) and 9.43 (US$1.03) for diastolic (CI 95% 19.90, 2.49). CONCLUSIONS: The reduction in blood pressure obtained may well justify the small incremental cost of the intervention. PMID- 11910715 TI - [Continuous oral hydration or with fractionated doses in acute diarrhea-induced dehydration in children]. AB - OBJECTIVE: To evaluate the safety and effectiveness of two oral rehydration techniques. MATERIAL AND METHODS: A randomized clinical trial was conducted at the oral rehydration unit of Hospital Infantil de Mexico "Federico Gomez", between September 1998 and June 1999. Forty patients five-year old and younger children, dehydrated due to acute diarrhea, were given oral rehydration solution (ORS) ad libitum (AL group); another forty patients received ORS in fractionated doses (FD group). Clinical characteristics were similar in both groups. Results are presented as means, standard deviations and medians, according the distribution of simple and relative frequencies. RESULTS: The mean stool output in the AL group was 11.0 +/- 7.5 g/kg/h; as compared to 7.1 +/- 7.4 in the FD group (p = 0.03). ORS intake, rehydration time, and mean diuresis values were similar in both groups (p > 0.05). Six patients in the AL group and five in the FD group had high stool output (> 10 g/kg/h), that improved after administration of rice starch solution. One patient in the AL group and two in the FD group had persistent vomiting that improved with gastroclisis. No patient required intravenous rehydration. CONCLUSIONS: These results suggest that ORS administration ad libitum under supervision, is a technique as safe and effective as the fractionated doses technique, for the treatment of dehydrated children due to acute diarrhea. PMID- 11910716 TI - Carriage of antibiotic-resistant pneumococci in a cohort of a daycare center. AB - OBJECTIVE: To define epidemiologic relationships to determine the prevalence and potential risk factors for nasopharyngeal colonization by antibiotic-resistant pneumococci, their serotypes and their antibiotic susceptibility patterns in children attending a daycare center (DCC). MATERIAL AND METHODS: A prospective cohort study was conducted among children (n = 53) attending the DCC at Hospital Infantil de Mexico Federico Gomez, which is staffed by 20 employees. Patients were enrolled in the study during a two-year period from September 1997 to September 1999. All the participants were followed prospectively, swabbing them every four months. The strains recovered were typed and screened for susceptibility to several antibiotics. The daycare records were reviewed also. Odds ratios and fisher's exact test: or chi square test of significance were computed from contingency tables as appropriate. Exact 95% confidence intervals were computed for odds ratios. Data analysis was performed using Epi statistics program version 6.04 a. RESULTS: Pneumococci were recovered from 45/53 of the infants at one or more visits. A total of 178 isolates were carried. The carriage rate was 47%. Only 7 adults acquired pneumococci during the study. Types 6, 14, 19 and 23 were prevalent and represented 77% of the total. Antibiotic-resistant strains were higher to penicillin and erythromycin. CONCLUSIONS: Children were frequent carriers of pneumococci, the rate of carriage was high in infancy and tended to decrease with age. The types commonly carried by children were the same as those causing invasive disease. There is a high proportion of carriers with antibiotic-resistant S. pneumoniae strains. Children who have had frequent antimicrobial courses are at particular risk. PMID- 11910717 TI - [Factorial validity of the Spanish adaptation of the Maslach Burnout Inventory General Survey]. AB - OBJECTIVE: To explore the factorial structure of a Spanish version of the "Maslach Burnout Inventory-General Survey" (MBI-GS). MATERIAL AND METHODS: A cross-sectional study was conducted in 1999 among 149 municipal policemen in Tenerife, Spain. A questionnaire was applied to collect data on Professional Efficacy, Cynicism, and Burnout. Data were analyzed using factorial analysis and principal components with Varimax rotation. RESULTS: Four factors had eigenvalues greater than I; Factor I grouped Professional Efficacy items, Factor II grouped three Cynicism items. Further analysis was conducted to limit extraction to three factors. The factorial solution replicated the distribution of items in the questionnaire manual. CONCLUSIONS: Study results show that the Spanish version of the questionnaire is valid and reliable. PMID- 11910718 TI - [Family life strategies and their relation with malnutrition in children under 2 years old]. AB - OBJECTIVE: To identify the role of family life strategies on malnutrition in children aged 6-23 months of age. MATERIAL AND METHODS: This case-control study was conducted in 1998 in the municipality of Teolocholco, State of Tlaxcala, Mexico, among families with children aged 6-23 months of age. The sample was conformed by 105 cases and 210 controls. Family life strategies were grouped into five types: family composition, means and distribution of family income, family and social networks, and life preservation strategies. Malnutrition was classified according to height for age. Data were analyzed using logistic regression to obtain odds ratios and 95% confidence intervals. RESULTS: Data were collected from 605 families, for a total of 445 controls and 160 cases. The predictive model included mother's schooling, overcrowding, time elapsed between childbirths, per capita monthly income, and time devoted to child-rearing activities. CONCLUSIONS: Family life strategies determine children's nutritional status; understanding the influence of the family on the children's health status is necessary to develop effective programs aimed at improving the nutritional status of children. PMID- 11910719 TI - [Exposure to group B Streptococcus among Mexican women in reproductive age]. AB - OBJECTIVE: To assess the prevalence of IgG antibodies against Group B streptococci (GBS) among women of reproductive age in Mexico. MATERIAL AND METHODS: Serum specimens were drawn from 15 to 40 year-old women, representative of all regions and socioeconomic levels of the country. The sample was randomly selected from Banco Nacional de Sueros (National Sera Bank); serum samples were collected during a national seroepidemiologic survey conducted in 1987-1988. The assays for standardization and for evaluation of seroprevalence were carried out at the Hospital de Pediatria del Centro Medico Nacional Siglo XXI (Children's Hospital) Instituto Mexicano del Seguro Social (IMSS) (Mexican Institute of Social Security) from January to November 1995. IgG antibodies against group B antigen were studied with an enzyme-linked immunosorbent assay (ELISA) developed in our lab. Group B antigen was produced and purified from the reference strain GBS 110. RESULTS: A total of 2669 serum samples were studied; 2405 were positive to anti-group B antigen IgG antibodies, for a seroprevalence of 90.2%. No differences in prevalence were found among the different age groups or among the different states of the country. CONCLUSIONS: The high seroprevalence of antibodies against GBS suggests that young women in Mexico are commonly exposed to GBS infection. PMID- 11910720 TI - [Re-emergence of hemorrhagic disease in newborns. Implications for its prevention]. AB - OBJECTIVE: To describe the occurrence of hemorrhagic disease of the newborn (HDN) at a tertiary care pediatric hospital of Morelos state. MATERIAL AND METHODS: A retrospective case series study was conducted between 1997-2000 at Hospital del Nino Morelense (Morelos State Children's Hospital), in 46 newborns aged under 12 weeks. Study subjects were referred from peripheral units with a diagnosis of HDN. RESULTS: The severe late-onset form of HDN was present in 91% of the cases. Fifty-two percent of childbirths were assisted by a physician and 48% by an empiric midwife. Application of vitamin K was unknown in 61% of cases, in 39% it was not applied and in 4% it was applied. The majority of infants presented severe symptoms due to intra-cranial bleeding, 11% died, and 41% had severe disease sequelae. CONCLUSIONS: Given the high prevalence of HDN in the State of Morelos, reproductive health programs should be reviewed and training programs intensified to promote the utilization of vitamin K by physicians and nurses for preventing this disease. PMID- 11910721 TI - The Chilean health system: 20 years of reforms. AB - The Chilean health care system has been intensively reformed in the past 20 years. Reforms under the Pinochet government (1973-1990) aimed mainly at the decentralization of the system and the development of a private sector. Decentralization involved both a deconcentration process and the devolution of primary health care to municipalities. The democratic governments after 1990 chose to preserve the core organization but introduced reforms intended to correct the system's failures and to increase both efficiency and equity. The present article briefly explains the current organization of the Chilean health care system. It also reviews the different reforms introduced in the past 20 years, from the Pinochet regime to the democratic governments. Finally, a brief discussion describes the strengths and weaknesses of the system, as well as the challenges it currently faces. PMID- 11910722 TI - [Transmissible spongiform encephalopathies]. AB - Transmissible spongiform encephalopathies (TSE) are a group of diseases which have received a lot of attention in recent years. The interest on these diseases has been stimulated by the appearance of bovine spongiform encephalopathy (BSE) and the new variant of Creutzfeldt-Jakob disease (nvCJD); the latter is likely to be acquired by ingesting contaminated beef. Until now 109 cases of nvCJD have been reported, most of them occurring in the United Kingdom. Some experts think that this is the beginning of a nvCJD pandemic. Deep knowledge of the mechanisms of transmission of TSE is needed to prevent the emergence of a TSE pandemic in humans. We address various aspects of TSE and discuss prevention methods of TSE in ruminants and humans. PMID- 11910724 TI - Mechanical ventilation in acute lung injury and ARDS. Tidal volume reduction. AB - Traditional mechanical ventilation practices used generous tidal volumes in patients with acute lung injury and acute respiratory distress syndrome (ALI/ARDS). This approach may have caused overdistention of aerated lung units, thus exacerbating lung injury in some patients. Several recent clinical trials of traditional versus lower tidal volume strategies in ALI/ARDS yielded disparate results. In the largest study, the lower tidal volume approach was associated with lower mortality and more ventilator-free days. This article reviews the rationale for tidal volume reduction in ALI/ARDS and the differences between the studies. Several different interpretations of the recent clinical trial results are addressed. PMID- 11910723 TI - [Cost of care of arterial hypertension and its impact on the budget assigned to health in Mexico]. AB - OBJECTIVE: To assess the medical care costs of hypertension and their impact on the health care expenditures and on Mexico's Gross National Product (GNP). MATERIAL AND METHODS: An ecological study was conducted from June to November 1999, at Instituto Mexicano del Seguro Social (Mexican Institute of Social Security, IMSS), in Monterrey, Nuevo Leon, Mexico. A random sample of medical charts of patients with hypertension was selected, to extract data on utilization of health services and unitary costs per care episode. The cost per care episode and per hypertensive patient was calculated by adjusting the unitary cost as a function of standard and extreme utilization of IMSS health services. The resulting figure was then projected to the total population of hypertensive patients and compared to the annual health care expenditures of Mexico. RESULTS: The annual cost per patient with hypertension was $1,067 in the standard scenario and $3,913 in the extreme scenario. The annual expenditures from hypertension corresponded to 13.95% of the budget allocated to health care and to 0.71%, of Mexico's GNP. These figures changed to 51.17% and 2.61% in the extreme scenario, respectively. CONCLUSIONS: The costs of hypertension medical care account for a good portion of healthcare resources. This problem should be analyzed by multidisciplinary health teams in search of more efficient medical care alternatives. PMID- 11910725 TI - Ventilator-associated pneumonia. Prevention, diagnosis, and therapy. AB - Ventilator-associated pneumonia remains the nosocomial ICU infection of greatest concern. The authors have summarized the clinical trials that have assessed specific strategies to prevent VAP and the current controversies regarding the diagnosis and therapeutic approach to this condition. Improvements in care of patients who are at risk for or who have developed VAP will depend on the judicious application of this information for individual patients. PMID- 11910726 TI - ARDS and the multiple organ dysfunction syndrome. Common mechanisms of a common systemic process. AB - The multiple organ dysfunction syndrome is a common but poorly understood complication of critical illness. Its evolution reflects the interactions of an acute, life-threatening insult, the response of the host to that insult, and the therapeutic measures instituted to restore normal homeostasis. Although the cellular mechanisms remain elusive, processes such as inflammation, microvascular thrombosis, apoptosis, and fibrosis and tissue repair contribute to its clinical expression. In the lung, these forces create the characteristic changes of ARDS; that common disorder, however, is better seen as one manifestation of a systemic process than as an isolated problem of the lung. Therapy, in the absence of a more sophisticated understanding of pathologic mechanism, is supportive. The growing recognition that iatrogenic factors contribute to the expression of MODS has highlighted the need for the clinician to be aware of the potential for harm inherent with every intervention. PMID- 11910727 TI - ARDS. Monitoring tissue perfusion. AB - A clinically feasible method for assessing regional splanchnic perfusion is still lacking. Methods used for research purposes demonstrate that the effects of current therapies on splanchnic perfusion are not predictable in intensive care patients with and without ARDS. Tonometry, laser Doppler flowmetry, and spectrophotometry have been used to assess splanchnic perfusion. Combining the available methods in different parts of the gastrointestinal tract may help assess splanchnic perfusion more accurately in the near future. PMID- 11910728 TI - Optimal peep in ARDS. Changing concepts and current controversies. AB - From many recently performed studies, it is clear that a criterion standard for determining the optimal positive end-expiratory pressure (PEEP) level in patients with acquired respiratory distress syndrome (ARDS) does not exist. What is evident and consistent, however, are several points such the optimal PEEP level ultimately represents a balance between regional areas of overstretching and regional derecruitment; higher levels of PEEP may be required early in ARDS, independent of oxygenation requirements; and the exact method for titrating PEEP in patients with ARDS remains to be determined. These points and others are delineated and discussed in this article. PMID- 11910729 TI - Cytopathic hypoxia. Is oxygen use impaired in sepsis as a result of an acquired intrinsic derangement in cellular respiration? AB - Several lines of evidence indicate that cellular energetics are deranged in sepsis, not by inadequate tissue perfusion but rather by impaired mitochondrial respiration; that is, organ dysfunction in sepsis may result from cytopathic hypoxia. If this concept is correct, the therapeutic implications are enormous. Efforts to improve outcome in septic patients by monitoring and manipulating cardiac output, systemic oxygen (DO2), and regional blood flow are doomed to failure. Instead, the focus should be on developing pharmacologic strategies (e.g., isoform-selective iNOS or PARP inhibitors) to restore normal mitochondrial function and cellular energetics. PMID- 11910730 TI - ARDS. The future. AB - Improving the course and outcome of patients with ARDS presents a considerable challenge. An important component of meeting this challenge is a more comprehensive understanding of the heterogeneous pathophysiology of ARDS and the biologic response of the individual patient. This understanding may be developed through the power of genomics and its related technology. In particular, it will be crucial to characterize the immunophenotypes of individual patients with ARDS. By understanding the immune status of a given patient at a given point in the disease process, physicians can consider manipulating proinflammatory systems more rationally, such as the complement and chemokine cascades, or the anti inflammatory arm of the immune system. Finally, a more refined molecular and genetic understanding of endogenous cytoprotective molecules and mechanisms, such as the heat shock response and HO-1, may provide further tools in the future armamentarium against ARDS. PMID- 11910731 TI - Effects of prone position ventilation in ARDS. An evidence-based review of the literature. AB - In summary, the many studies done on PPV show that the technique improves oxygenation most of the time. The mechanisms behind this effect are probably numerous and have yet to be elucidated completely. In addition, PPV is a safe procedure that rarely worsens a patient's respiratory status or causes other complications and is thus a welcome additional therapeutic option when treating patients with ARDS. Despite the recent large, randomized, controlled trial showing no improvement in mortality rate or organ dysfunction overall, there is evidence suggesting that PPV may be of most benefit in more severely ill patients. Further studies will be useful. PMID- 11910732 TI - Inhaled nitric oxide in ARDS. AB - The role of nitric oxide (NO) in numerous physiologic systems only recently has been discovered. When used as a gas, inhaled NO (iNO) has many unique properties that cause immediate improvements in pulmonary hemodynamics and oxygenation. Acute benefits in physiologic parameters have been demonstrated in numerous studies of iNO in acute respiratory distress syndrome (ARDS), but recent randomized controlled trials have failed to show improvement in outcome. The addition of other treatments that prolong or enhance the affect of iNO or its use with other ventilator modalities such as prone positioning or high-frequency ventilation offer hope that iNO may be beneficial in select groups of patients. PMID- 11910733 TI - New management strategies in ARDS. Immunomodulation. AB - Acute respiratory distress syndrome is a common condition among the critically ill and is associated with high morbidity and mortality [table: see text] rates. Improved understanding of the underlying inflammatory pathogenetics has encouraged the search for strategies that, by modifying this immune response, can improve outcome for this group of patients. Some agents are obviously anti inflammatory. Others have been used primarily for other purposes; their immune effects are incidental, but no less important. Although immunomodulatory strategies have been discussed for many years, they now are beginning to show positive results, as in the study using activated protein C. Most patients with ARDS die with ARDS, rather than from ARDS. The approach to treatment must not be lung-limited but must take into account the systemic effects of the inflammatory response. The complex nature of the syndrome makes it likely that no single agent will provide the long-desired cure. Rather, it is probable that an individual patient will require a combination of several agents or different agents at different times during the disease process (Table 1). Mortality rates from ARDS already are beginning to fall with improved nutritional and ventilatory support. Positive results from trials using immunomodulatory agents are being reported, and soon such agents will form part of the routine management of patients with ARDS, further improving the outlook for these patients. PMID- 11910734 TI - Corticosteroids in ARDS. An evidence-based review. AB - In general, a rule for corticosteroids in preventing or relieving the acute respiratory distress syndrome (ARDS) has yet to be established, although these drugs are indicated for conditions such as Pneumocystis carinii pneumonia. High dose corticosteroids have not been shown to reduce mortality through their anti inflammatory properties when given early to patients with sepsis, septic shock, or ARDS. Corticosteroids have been shown, however, to reduce mortality in patients with late ARDS only in one small, inconclusive study. More recent investigators have focused on the usefulness of low-dose corticosteroids in reducing mortality in patients with sepsis or septic shock who may have relative adrenal insufficiency, but these studies also are inconclusive, and it is unclear that low-dose corticosteroids affect the development of ARDS in these patients. PMID- 11910735 TI - New therapies for adults with acute lung injury. High-frequency oscillatory ventilation. AB - High-frequency oscillatory ventilation seems theoretically ideal for the treatment of patients with ARDS, allowing adequate oxygenation and ventilation to be maintained without causing further damage to the already injured lung. High frequency oscillating ventilation also seems a sound strategy for improving oxygenation in patients who are no longer responding to conventional mechanical ventilation. Currently, HFOV should be used in the adult ICU as one of many ancillary therapies available for the treatment of extremely ill, hypoxemic patients with ARDS. Future research may define the role of HFOV as a more routine strategy for preventing VALI in this patient population. PMID- 11910736 TI - [Pharmaceutical importance of Allium sativum L. 2. Antibacterial effects]. AB - The communication summarizes mainly newly obtained experimental findings confirming the antibacterial effect of garlic preparations (powders, extracts, juice, essential oil, oil macerate) and their individual components. It also reports the effectiveness of substances newly isolated from the oil macerate (iso E-10-devinylajoene, Z-10-devinylajoene, and three, or five thiosulfinates). The effect on the tested bacteria included here is, according to new evidence, indisputable. The paper has purposefully excluded the action of garlic against the widely distributed bacteria Helicobacter pylori, which causes chronic gastritis, gastric and duodenal ulcers. The findings concerning this matter will be published in the following communication. PMID- 11910737 TI - [Drug chirality]. AB - Chirality is a stereochemical phenomenon in the area of drugs, influencing pharmacodynamic, pharmacokinetic, and biotransformation processes. On an example of known drugs, the present paper describes the relations of chirality to drug efficacy, safety, toxicity, manifestations of drug interactions, and some other factors with a feedback action on the chirality-drug relationship. PMID- 11910738 TI - [Advances in the development of new antimycobacterial agents. The alkylsulfanyl group--a pharmacophore for antimycobacterial activity]. AB - Tuberculosis ranks among the most widely spread infectious diseases and the need to find new antituberculotics has become more and more urgent. The present review paper aims at the compounds containing an alkylsulfanyl group which can be considered the pharmacophore of antituberculotic activity. The alkylsulfanyl group is bound to various heterocycles, condensed systems, and there is also a bond to the aliphatic structure. The activities of some compounds discussed in the resent paper are comparable with those of the antituberculotics in current use and therefore they could be introduced into practice, or at least suggest further direction of development. The present survey is based on the journal Chemical Abstracts. PMID- 11910740 TI - [Liberation of potential local anesthetics from colloidal dispersions]. AB - Evaluation of pharmaceutical availability of drugs from topical preparations is usually aimed to evaluate the capability of the dosage form to release the drug for its therapeutic action. Permeation cells have been used for this purpose, employing solid membranes including cellophane. Within a study of phenylcarbamic acid derivatives with local anaesthetic effect, the present paper evaluates the dosage form for topical administration containing potential drug XIX M, the chemical name N-[2-(2-heptyloxyphenylcarbamoyloxy)-ethyl]-morpholinium chloride and potential drug XXII B, the chemical name N-[2-(2-octyloxyphenylcarbamoyloxy) ethyl]-dimethylammonium chloride. The selected auxiliary substance in the formulation of the dosage form was a cellulose derivative--methylcellulose in increasing concentrations from 0.5 to 1.5%. On the basis of the obtained results it can be stated that the examined polymer methylcellulose with increasing concentration decelerated the release of both potential drugs approximately in the same way and that it can be used in practice to prolong the local anaesthetic effect. PMID- 11910741 TI - [Effect of magnesium stearate on the tensile strength of tablets made with the binder Prosolv SMCC 90]. AB - The present paper evaluated the tensile strength of tablets made from the dry binder Prosolv SMCC 90 and the influence of three concentrations of the lubricant magnesium stearate on the tensile strength of tablets manufactured from this material. The results were compared with the same evaluation in Avicel PH 102. The tested concentrations of the stearate were 0.4, 0.8 and 1.2%. The tablets were compressed by three press powers (3, 3.5, and 4 kN). On the basis of obtained results it can be stated that under the same press powers Prosolv SMCC 90 alone yields stronger compacts than Avicel PH 102. From the viewpoint of decreased strength of compacts by adding magnesium stearate, the dry binder Prosolv SMCC 90 is much less sensitive than Avicel PH 102. In Avicel PH 102 a marked decrease in tensile strength was recorded with an addition of 0.4%, which was not observed with Prosolv SMCC90. A more significant decrease in the strength of compacts was shown by the substance not until a stearate concentration of 0.8%. The highest employed stearate concentration of 1.2% decreases the tensile strength of tablets made from Prosolv SMCC 90 in the press powers of 3.5 and 4 kN two times less than the tensile strength of the compacts from Avicel PH 102. PMID- 11910742 TI - [Effect of CrCl3 on the in vitro production of flavonoids in a culture of Ononis arvensis L]. AB - The paper investigated the effect of the abiotic elicitor chromium trichloride on the production of flavonoids in the callus and suspension cultures of Ononis arvensis L. after 12; 24; 48; 72, and 168-hour administration. The tested elicitor positively influenced the formation of flavonoids. A statistically significant increase in the flavonoid production in the callus culture was recorded with the use of a chromium trichloride solution in the concentration c1 (6.32 x 10(-4) mol/l) after 24; 48; 72, and 168 hours, in the concentration c2 (6.32 x 10(-6) mol/l) after 24; 48, and 72 hours, and in the concentration c3 (6.32 x 10(-8) mol/l) after 12; 24, and 48 hours. In the suspension culture, the flavonoid production was significantly increased by the concentration c1 (6.32 x 10(-4) mol/l) after 12 and 72 hours, the concentration c2 (6.32 x 10(-6) mol/l) after 12; 24; 48, and 72 hours, and the concentration c3 (6.32 x 10(-8) mol/l) after 12; 24; 48, and 72 hours. The maximal increase in the flavonoid production by 98% took place in the callus culture after elicitation with a chromium trichloride in the concentration c3 (6.32 x 10(-8) mol/l) after 48 hours of administration, and in the suspension culture after elicitation with CrCl3 in the concentration c2 (6.32 x 10(-6) mol/l) after 12 hours by 100%. PMID- 11910743 TI - [Effect of precursors on the production of coumarins in a suspension culture of Angelica archangelica L]. AB - The effect of potential precursors (cinnamic acid, phenylalanine, tyrosine) in three concentrations (0.01; 0.1; 1 mmol/l) on the growth of the culture and coumarin production was investigated in the suspension culture of Angelica archangelica L. The cultures were cultivated under constant illumination (3500 lux) and in the dark. The growth of the culture in the dark was not affected by the precursors, cinnamic acid in a concentration of 1 mmol/l exerted toxic action. The growth of the culture cultivated under constant illumination was stimulated by tyrosine in a concentration of 0.1 and 1 mmol/l, and phenylalanine in a concentration of 0.1 mmol/l; phenylalanine and cinnamic acid in a concentration of 1 mmol/l inhibited the growth of the culture. In the cultivation in the dark, coumarin production was increased by phenylalanine in a concentration 0.01 mmol/l and by tyrosine in a concentration of 1 mmol/l. In the cultivation under constant illumination, coumarin production was decreased by the action of cinnamic acid, was not affected by tyrosine, and phenylalanine (0.01 and 0.1 mmol/l) increased it in comparison with the culture without precursors. PMID- 11910744 TI - [Penicillin G acylase--synthesis, regulation, production]. AB - Penicillin G acylase (PGA) is one of very important industrial enzymes used in the production of polysynthetic beta-lactam antibiotics. This enzyme catalyzes the hydrolysis of the amidic bond of penicillin G with the development of 6 aminopenicillanic acid which serves as the initial substance for the production of semisynthetic penicillins. In the strain Escherichia coli W ATCC 11105 and ATCC 9637, PGA is coded by the pga gene on the chromozome and synthesized as the pre-pro-PGA (pp PGA) precursor, which is transported, with probable participation of the chaperon system, to the periplasmatic space of the cell. Here after a series of proteolytic reactions the active enzyme PGA develops, consisting of two subunits alpha and beta. Expression of the pga gene is subject to several regulatory mechanisms: temperature repression, catabolic repression by glucose, repression by oxygen, and induction by phenylacetic acid (FOK). The formation of active PGA is also influenced at the post-translation level, where an important role is played by intracellular proteolytic reactions and the transport system of pre-pro-PGA across the cytoplasmatic membrane. The chromozomal area of the pga gene of the E. coli W strain was employed for the construction of many recombinant plasmids. These plasmids served to transform suitable host strains, some of which are now used in industry as highly productive microorganisms. PMID- 11910745 TI - Development of an RNFA program: a Newfoundland and Labrador adventure. AB - This inquiry was to determine the feasibility of creating a nursing role for first assisting in the cardiac operating room in Newfoundland and Labrador. A committee was struck to review the role as it existed in the United States and Canada. Following this, the committee gained approval from appropriate professional associations and began developing the program. The result is that three nurses are currently working as RNFA's in Cardiac Surgery and two nurses are piloting a RNFA Program for General Surgery. PMID- 11910746 TI - Computerized O.R. scheduling: is it an accurate predictor of surgical time? AB - The goal of this project was to determine whether a standardized surgical time, generated by the Operating Room Information System (ORIS), could be used as an accurate predictor of actual surgical time. Utilizing retrospective, quantitative data from the ORIS database, frequency distributions by surgical speciality, were completed. Chi-square analysis was applied to determine the significance of the frequency distributions. The study outcome indicates that ORIS computer generated procedure times were not an accurate predictor of actual surgical time. Further follow-up will be required to determine if alternate scheduling methodologies would lead to higher accuracy rates. PMID- 11910747 TI - [The development of nanoparticles on DNA isolation and purification]. AB - Magnetic nanoparticle is a new solid-support of affinity chromatography. The particle size is small and it has super-paramagnetism. It has large surface area and it can be endowed many reaction groups such as streptavidin, antibody or DNA fragments. The target DNA can be separated in magnetic field. The magnetic nanoparticle is applied in the biomolecular field gradually and it has a very broad prospect of appliance. PMID- 11910748 TI - [Progress in telomerase and the inhibitors]. AB - In cells division chromosome length is shorten for the ending DNA couldn't be replicated completely, and the loss of telomere DNA will lead to senescence and death. Activation of telomerase can elongate telomere length and maintain gene stability. Up-regulation of telomerase is considered to be responsible for immortalization and carcinogenesis. It plays an important role in cell-span and cell division. The telomerase inhibitors will become efficient drugs in tumor therapy. PMID- 11910749 TI - [Advances in in vitro refolding of inclusion body proteins]. AB - Overexpression of recombinant proteins in E. coli often results in formation of insoluble, inactive inclusion bodies. These inclusion bodies, which contain the recombinant proteins in a highly enriched form, can be isolated by solid/liquid separation. After solubilization, active proteins can be generated through an appropriate refolding process. Within the last decade, specific strategies and methods have been developed for preparing active recombinant proteins from inclusion bodies. Recent developments in renaturation procedure include the inhibition of aggregation during refolding by the application of low molecular weight additives and matrix-bound renaturation techniques. PMID- 11910750 TI - [New progress of serial analysis of gene expression]. AB - As an efficient tool that has been developed recently, serial analysis of gene expression (SAGE) allows the qualitative and quantitative analysis of a large number of transcripts. It can define the transcripts at relatively low levels and characterize the genomic expression near completion. In addition, it provides insights into the timely and orderly expression of genes by comparing the profiles constructed for a pair of cells that are kept at different conditions, thus identifying a set of novel genes. In this review, the newest progress of SAGE's application and research is mentioned in details with its original method and principle outlined. PMID- 11910751 TI - [The application of genetic engineering to the petroleum biodesulfurization]. AB - The developed course and reaction mechanisms of petroleum biodesulfurization were introduced. The recent development of genetic engineering technology, which used in desulfuration strain's construction, reconstruction and other fields, was summarized emphatically. Its current research situation internal and overseas and the developing prospect were simply analyzed, and our research designs were submitted. PMID- 11910752 TI - [A fusion protein of rotavirus VP6 and cholera toxin B subunit: expression in Escherichia coli and analysis of biological activities]. AB - Rotavirus infection is a major cause of dehydrating diarrhea in infants worldwide. The non-toxic cholera toxin B subunit(CTB), known as an immunomodulatory carrier, might help to stimulate mucosal immune response when coupled to rotavirus antigens in oral immunization. Here we report for the first time the construction of a translational fusion of CTB gene 5' to the VP6 gene of a human rotavirus A(field strain T114), and expression of the CTB-VP6 fusion protein in E. coli BL21(DE3). The expressed fusion protein has a molecular weight of 56 kD, as expected, and accounts for about 15% of the total E. coli protein. Western blottings with the hyperimmune serum against rotavirus strain WA and the antibody against cholera toxin indicated that the fusion protein retains the antigenicity identical to the native CTB and VP6. The GM1-ELISA analysis proves that the renatured CTB-VP6 has strong affinity for GM1 ganglioside. PMID- 11910753 TI - [The inhibitory effects of hepatocyte targeting pH-sensitive liposome mediated phosphorothioate antisense oligonucleotide on gene expression controlled by HCV 5'NCR]. AB - Antisense oligodeoxynucleotide is a negative charged macromolecule and hence is difficult to penetrate cell membrane and liable to degradation. To increase the effective concentration of antisense drugs in the target cells, a hepatocyte targeting liposome directed to asialoglycoprotein receptors exclusively expressing on the hepatocyte membrane was designed and prepared based on the receptor-mediated gene transfer. In order to accelerate endosomal exit of nucleic acid drugs, the liposomal formulation with pH-sensitive property was adopted. The hepatocyte-targeting and pH-sensitivity of liposome were analyzed by galactose receptor competitive inhibition and hemolysis of chicken red blood cell. Antisense oligodeoxynucleotide, HCV363 against HCV 5'NCR, was delivered via prepared liposome to transgenic cell HepG2.9706 and evaluated for its inhibitory effect on luciferase expression controlled by HCV 5'NCR in HepG2.9706 using Luciferase Assay System The results showed that different concentrations (10, 20, 30 mmol/L) of galactose solutions reduce the delivery effects of liposome to some extend that were up to saturation when the concentrations of galactose solution exceed 20 mmol/L. Prepared liposomes mixed with chicken RBC are put into PBS buffers with different pH values(4.0-8.0), it was observed that the amount of heme is greatly released in acidic PBS (pH < 6) due to the fusion of liposome and RBC membranes. Liposome-mediated HCV363 has dose-dependent inhibitory activities on luciferase expression controlled by HCV 5'NCR in HepG2.9706 and the inhibitory rate is 86% at a concentration of 1.0 mumol/L. In conclusion, the liposome is proven to be a hepatocyte-targeting pH-sensitive delivery system that can increase the pharmaceutical effects of antisense drugs. PMID- 11910754 TI - [Cloning and expression of heptad repeat regions of the Newcastle disease virus fusion protein]. AB - Two heptad repeat regions (HR1, HR2) from F clone of the virulent and avirulent NDV were cloned, expressed with vector pGEX-6p-1 in E. coli BL21 (DE3), and purified the fusion protein by Glutathiones Sepharose 4B Column. After cleaved by precission protease, the desired protein was purified by Glutathione Sepharose 4B Column and high temperature. Purified HR1 and HR2 were analyzed by Mass spectrum, the results shows that the Molecular Weights of HR1 and HR2 of virulent NDV are 7103 and 6301, and the Molecular Weights of HR1 and HR2 of avirulent NDV is 7107 and 6309. PMID- 11910755 TI - [Studies on bioconversion conditions and stereoselectivity of Arthrobacter K1108 hydantoinase]. AB - The bioconversion conditions of hydantoinase of Arthrobacter K1108 were studied. It was shown that the optimum temperature of the enzyme is 55 degrees C, and the optimum pH is 7.0. The enzyme can be activated by Co2+ and Fe2+, while inhibited by Ca2+. The optimal substrate of the hydantoinase is 5-benzylhydantoin, while 5 phenylhydantoin and 5-indolylmethylhydantoin cannot effectively digested, showing a high specificity on the substrates. An investigation on the hydantoinase stereoselectivity mechanism showed that the N-carbamoylamino acid hydrolase is stereoselective but the hydantoin hydrolase is not. PMID- 11910756 TI - [Study of enzyme immunosensor immobilized by regenerated silk fibroin]. AB - The author studies the enzyme immunosensor made up of graphite electrode and protein film of regenerated silk fibroin which is employed to immobilized antigen (rabbit IgG). IgG will be recognized and combined by antibody (goat-anti-rabbit IgG-HRP). After enlarging the signal of the combination of antigen and antibody by H2O2, the concentration of IgG is able to be measured by testing the electrode potential directly. The Enzyme Immunosensor is sensitive when measuring IgG and the detection limit of 1.2 x 10(-10) mol/L is found. It shows linear response over the concentration range of 4.1 x 10(-7)-1.2 x 10(-10) mol/L. Electrophoresis cuts down the time of antibody-antigen interaction from 90 min to 30 min. The response time is 15 s. Enzyme immunosensor with better stability and higher sensitivity can be used extensively in clinical diagnosis, medical and environmental studies, HLA molecular diagnosis and so on. PMID- 11910757 TI - [Expression, purification and identification of human matrix metalloproteinase 2]. AB - The expression sequence of matrix metalloproteinase-2 (MMP-2) has been obtained by PCR amplifying, restriction enzyme cut and sequencing analysis demonstrate that the sequence is correct. The recombinant expression plasmid pPIC9/MMP-2 containing MMP-2 is constructed and transformed the yeast Pichia pastoris. Recombinant matrix metalloproteinase-2 protein was expressed in Pichia pastoris in great deal after induction by methanol. The purity of the recombinant MMP-2 filtrated through Sephacryl S-200 reached to electrophoresis purity. With the ability to degrade gelatin and IV type collagen, recombinant MMP-2 has the similar substrate specificity with natural MMP-2. The recombinant MMP-2 with 50 kD molecular weight is smaller than natural MMP-2, which suggested degradation occurred to it. PMID- 11910758 TI - [Studies on the expression condition of human trefoil factor 3 in Pichia pastoris]. AB - In order to enhance the expression level of human trefoil factor 3 (hTFF3) in Pichia pastoris, we optimized the transformant growth conditions in shake flasks including carbon sources in growth medium, inoculation ratio, methanol concentration, pH rotation speed and inducing time. The transformant could grow on the glucose to OD600 5.0 after 14 hours inoculation. The best inoculation ration of 100 mL growth medium to the induction medium was 1:1. The expression level of dimeric human trefoil factor 3 induction with 1% methanol for 48 hours at pH 6.0, agitation speed 240 r/min could reach 20 mg/L with OD600 15. The protein was expressed in 5-liter fermentor with 2% methanol induction for 32 hours, finally the cell density reached OD600 120. 100 mg/L of recombinant hTFF3 was obtained in the supernatant. PMID- 11910759 TI - [Enhanced immunogenicity of plasmid encoding polyprotein gene of infectious bursal disease virus by co-administration of chicken interleukin 2 (IL-2)]. AB - Chicken interleukin 2 (IL-2) is one of important nonmammalian cytokines isolated recently. The influencing of IL-2 on immunogenicity of DNA vaccine was examined using infectious bursal disease virus as a model. The IL-2 cDNA of Xiaoshan chicken and the polyprotein gene of IBDV-ZJ2000 were amplified by RT-PCR, cloned, sequenced and inserted into the control of CMV promoter and enhancer of pCI vector. 14-day-old chickens were vaccinated intramuscularly with DNA vaccine, two weeks later, they were boosted with DNA, and two weeks post boost, they were challenged with virulent IBDV. The results showed that protective responses and neutralization antibody responses of DNA vaccine co-administrated with chicken IL 2 were much higher than those of injected with DNA vaccine alone. Furthermore, the T lymphocyte proliferation response of peripheral blood, thymus and spleen, and the B lymphocyte proliferation response of bursa induced by DNA vaccine can be significantly enhanced by chicken IL-2. These results obviously indicated that chicken IL-2 was a strong adjuvant which can significantly enhance the immunogenicity of IBDV DNA vaccine. PMID- 11910760 TI - [Cloning of HAL1 gene and characterization for salt tolerance tomato]. AB - The HAL1 gene was cloned by PCR strategy and confirmed by sequencing. Its open read frame is 879 bp, encoding a peptide of 294 amino acids (32 kD Protein). A chimaeric construct of HAL1 and Npt II (neomycin phosphotransferase II) was constructed and introduced into commercial cultivars of tomato (Zhong SU No. 5: Lycopersicon escullentum) by Agrobacterium tumefacien-mediated gene transformation. Transformants were selected for their ability to grow and root on media containing kanamycin. Transformation was confirmed by analysis of PCR, Southern blot and RT-PCR. The salt tolerance of transgenic tomato is evaluated by comparing the fresh weight, dry weight, Na+, K+ content of transgenic tomato and control tomato. It is concluded that the over-expressing of HAL1 in tomato could enhance the salt tolerance of the transgenic tomato. PMID- 11910761 TI - [Cloning and structure analysis of a restriction and modification system, LlaBIII from Lactococcus lactis subsp. cremoris W56]. AB - A 22.4 kb naturally occurring plasmid pJW566, isolated from L. lactis W56, was found to encode an R/M system named LlaBIII. The LlaBIII R/M system was isolated on a chloramphenicol resistant derivative of plasmid pJW566, resulting in a plasmid pJK1. Subcloning analysis showed that the LlaBIII determinant was located on a 5 kb HindIII-Sph I fragment. The fragment was sequenced. It contained a single open reading frame (ORF), corresponding to a protein of 1584 or 1576 aa. In the deduced amino acid sequence seven helicase motifs characteristic of endonuclease type I and type III and a conserved catalysis motif X in the R subunits of type I R/M systems were located in the N-terminus, followed by four conserved motifs found in DNA N6-adenine methyltransferases. The C-terminus of the deduced amino acid sequence showed no homology to known R/M systems. Therefore, this polypeptide encoded by LlaBIII is a multifunctional protein possessing putative DNA recognition, methylation and restriction activities. PMID- 11910762 TI - [Construction of cDNA library of Eimeria tenella sporulated oocysts]. AB - A lambda ZAP express cDNA library was constructed using mRNA from Eimeria tenella sporulated oocysts. Total RNA was isolated by the TRIzol from Eimeria tenella sporulated oocysts, mRNA was further purified through oligo(dT)-cellulose columns. The first-strand cDNA was synthesized by using MMLV reverse transcriptase with oligo(dT)18 primers containing Xho I restriction site. After the second strand cDNA replacement synthesized, the uneven termini of the double stranded cDNA were filled in with cloned Pfu DNA polymerase and EcoR I adapters were ligated to the blunt ends. Then the double-strand cDNA was digested with Xho I restriction enzyme. The fragments of 0.5 kb-4 kb were collected by agarose gel fraction method. After ligation of the cDNA with the lambda ZAP Express vector, the cDNA library was packaged using Gigapack III Gold Packaging extract. According to the phage plaques bright selection, the cDNA library contained 6 x 10(6) clones and the titer of the amplified library was 1 x 10(11) pfu/mL. By using PCR identification, the cDNA library contained approximately 96% recombinant phages. PMID- 11910763 TI - [Establishment of detective systems for GL-7ACA acylase expression]. AB - Glutaryl-7-amino cephalosporanic acid (GL-7ACA) acylase catalyzes the conversion of GL-7ACA to 7-amino cephalosporanic acid (7-ACA). The product 7-ACA is a starting compound for semi-synthetic cephalosporin antibiotics in industry. In order to detect the expression and specific activity of protein-engineered GL 7ACA acylase accurately, two useful detective systems for its expression has been established, in which reporter genes xylE and lacZ were fused to the downstream the GL-7ACA acylase gene acy respectively and the activity of catechol dioxygenase or beta-galactosidase could indicate the amount of acy expression. PMID- 11910764 TI - [Bone-inducing activity of human bone morphogenetic protein-2 102 peptide]. AB - To analyze the bone-inducing activity of C terminal of hBMP-2 and get a new recombinant product of hBMP-2, the gene encoding 102 aa of hBMP-2 mature peptide C terminal was cloned and expressed in E. coli and the first Cys was mutated with Ser. The fragments encoding the target peptide were amplified and cloned into heat-inducible expression vector pDH and transformed into E. coli DH5 alpha. After induction, a new protein bond appeared on the SDS-PAGE. The expressed products amounted to 30% of the total bacterial protein, which existed in the form of inclusion body. The products of bacterial lysates were purified through the ion-exchange chromatography. The denatured proteins were dialysed and diluted directly into the refolding buffer. The renatured products were implanted into mouse thigh muscles to analyze their bone-inducing activity respectively. The results of histological assay showed that the 102 peptide of hBMP-2 could ectopically induce formation of bone, while the mutated 102 peptide of hBMP-2 could not. It suggested hBMP-2 102 peptide still had bone-inducing activity. The first Cys of hBMP-2 mature peptide might be necessary for integrity of three pairs of disulfide bond, and also essential for bone-inducing activity of hBMP-2. PMID- 11910765 TI - [Decolorization and isolation of recombinant hirudin expressed in the methylotrophic yeast Pichia pastoris]. AB - Schemes for decolorization of broth and isolation of recombinant hirudin from the fermentation supernatants of recombinant yeast Pichia pastoris were developed to remove a great deal of non-desired proteins and coloring matters. Based on the stability of hirudin, heating is desirable to pre-treat the fermentation broth. Several different column chromatographies, including anion-exchange, cation exchange, hydrokyapatite adsorption, hydrophobic interaction, gel filtration and their combination were investigated on the efficiencies of decolorization and isolation. Among these methods, the combination of cation-exchange and anion exchange was found to be a success in both decolorization and isolation of hirudin. PMID- 11910766 TI - [Effects of medium and culture conditions on polysaccharide synthesis by suspension cell culture of Gardenia jasminoides Eills]. AB - The effects of medium and culture condition on polysaccharide synthesis by suspension culture of Gardenia jasminoides Eills were studied. The results show that B5 was the optimum medium, 5-10 days was suitable for subculture periods, and an inoculum of 80 g/L wet cell was better to cell growth and polysaccharide accumulation. For carbon source, it was better to use glucose than sucrose on cell growth, but due to the higher price of glucose, 45 g/L compound carbon source combined sucrose with glucose (1:1) was the optimum. The effect of different kinds of nitrogen source on cell growth and polysaccharide synthesis was not so big, and 40-50 mmol/L nitrogen was optimum, lower concentration of nitrogen source could inhibit the synthesis of polysaccharides. In addition, by controlling the harvest time of the polysaccharide in the suspension culture, the accumulation of yellow pigment could be prevented, which is easier to the polysaccharide purification. PMID- 11910767 TI - [Analysis of parameters for optimum of ergosterol fermentation]. AB - Parameters in the process of ergosterol fermentation are studied. The relationship between biomass, ergosterol content in Saccharomyces cerevisiae and parameters such as DO, pH, OUR, glucose concentration, are discussed. Because of its good manipulation in yeast fermentation process, Do can thus serve as an effective control parameter. DO at about 12% (+/- 1%) can enhance the total yield of ergosterol considerably. PMID- 11910768 TI - [Effects of glucose on growth, metabolism and EPO expression in recombinant CHO cell cultures]. AB - The maximum viable cell density almost kept constant in the experimental range of initial glucose concentrations from 8.9 to 49.6 mmol/L in the batch cultures of recombinant CHO cells. The yield coefficient of lactate to glucose increased with the initial glucose concentrations from 8.9 up to 17.9 mmol/L, and then kept constant with above 17.9 mmol/L. There was little effect of glucose on glutamine metabolism. It was also demonstrated that the accumulative concentration of EPO increased with the increase of glucose concentration from 8.9 to 17.9 mmol/L, and decreased with its further increase from 17.9 to 49.6 mmol/L. It is showed that there is an optimum glucose concentration at which the EPO express of cultured recombinant CHO cells is on the peak. PMID- 11910769 TI - [Isolation, purification and renaturation of recombinant-DNA-derived porcine somatotropin]. AB - Large scale abstraction and isolation of bacterially synthesized, recombinant-DNA derived, porcine growth hormone (r-pST) is described. The r-pGH is found in genetic engineering E. coli as the form of inclusion bodies. Pellet fraction which were mainly inclusion bodies, after cell breakage and centrifugation, were collected. Cell envelope components, such as protein, lipid, endotoxin and nucleic acids are selectively removed from the pellet fraction by an EDTA/lysozyme/deoxycholate extraction. Inclusion bodies were dissolved using 6 mol/L guanidine/HCl and air oxidation is then carried out in the presence of the guanidine/HCl. The Guanidine/HCl protein mixture were diluted by renaturation solution. Guanidine/HCl were removed by dialysis and then correctly refolded, oxidized r-pGH were obtained. Injection experiment of hypophysectomized rats proved r-pST with high native bioactivity was obtained. PMID- 11910771 TI - [Compound RSA 11 beta-hydroxylation with Curvularia lunata]. AB - Compound RSA 11 beta-Hydroxylation of Curvularia lunata was studied. The composition of hydroxylation products was analyzed. It was found that the conversion rate with protoplasts was increased conspicuously, the construction and quantity of hydroxylation products was not alteration, but the percentage was changed greatly. The 14 alpha-OH percentage of protoplasts was 3.03-fold than that of mycelia, the 11 alpha-OH percentage was decreased 60%, and the conversion ratio of 11 beta-OH was more 1.29-fold than that of the mycelia. It affected the percentage of hydroxylation products when 2 mg/mL polyoxins were added into the mycelia conversion system. PMID- 11910770 TI - [Effects of metal ions on gamma-poly (glutamic acid) synthesis by Bacillus licheniformis]. AB - In the course of gamma-poly (glutamic acid) gamma-PGA fermentation, metal ions K+, Mg2+, Fe3+, Ca2+, Mo6+, Mn2+, Co2+ and Zn2+ in the medium have certain effects on the synthesis of gamma-poly(glutamic acid). Excess or lack of K+, Mg2+ and Fe3+ results in reduced yield of gamma-PGA. It was found that the gamma-PGA synthesis by Bacillus licheniformis was promoted obviously by Ca2+ and Mo6+. Synthesis and stereochemical composition of gamma-PGA was greatly regulated by Mn2+. gamma-PGA was not produced without Mn2+ in medium, and with the increase of Zn2+ concentration the yield of gamma-PGA and the proportion of D-glutamic acid in the peptide increase. Regulative effect of Co2+ and Zn2+ was almost the same as that of Mn2+, thus the combination of Mn2+, Co2+ cannot enhance gamma-PGA synthesis and affect stereochemical composition of gamma-PGA. Based on the experimental date, an appropriate formulation of metal ions in the medium for gamma-PGA production was obtained. PMID- 11910772 TI - [Information is prevention]. PMID- 11910773 TI - [A resource for the gravely ill, too]. PMID- 11910774 TI - [Good information essential for satisfaction]. PMID- 11910776 TI - [Another child, how does one face it?]. PMID- 11910775 TI - [Let's say what we have to say]. PMID- 11910777 TI - [An accompaniment for negotiating for the long term]. PMID- 11910779 TI - [Too often forgotten]. PMID- 11910778 TI - [Maintaining autonomy is a priority]. PMID- 11910780 TI - [Self control is its own reward]. PMID- 11910781 TI - Influenza. PMID- 11910783 TI - Receiving the torch in the era of sexology's renaissance. PMID- 11910782 TI - Infected areas as at 7 March 2002. PMID- 11910784 TI - Dermatoglyphics, handedness, sex, and sexual orientation. AB - Both handedness and dermatoglyphic asymmetry reflect early, prenatal influences and both have been reported to be associated with male sexual orientation; handedness has been related to female sexual orientation as well. Neurohormonal and developmental perturbation are two competing hypothesis that attempt to explain these connections. We attempted to replicate these associations and to extend dermatoglyphic asymmetry findings to women. Dermatoglyphic directional asymmetry and fluctuating asymmetry were unrelated to sexual orientation. Homosexual women, but not homosexual men, had highly significant increases in non right-handedness compared with same-sex heterosexual controls. Although this pattern of results does not allow resolution of the two competing models, it does lend additional support to a biological basis of sexual orientation. PMID- 11910785 TI - Differences in finger length ratios between self-identified "butch" and "femme" lesbians. AB - There is indirect evidence that heightened exposure to early androgen may increase the probability that a girl will develop a homosexual orientation in adulthood. One such putative marker of early androgen exposure is the ratio of the length of the index finger (2D) to the ring finger (4D), which is smaller in male humans than in females, and is smaller in lesbians than in heterosexual women. Yet there is also evidence that women may have different sexual orientations at different times in their lives, which suggests that other influences on female sexual orientation, presumably social, are at work as well. We surveyed individuals from a gay pride street fair and found that lesbians who identified themselves as "butch" had a significantly smaller 2D:4D than did those who identified themselves as "femme." We conclude that increased early androgen exposure plays a role in only some cases of female homosexuality, and that the sexual orientation of "femme" lesbians is unlikely to have been influenced by early androgens. PMID- 11910786 TI - Relationships among childhood sex-atypical behavior, spatial ability, handedness, and sexual orientation in men. AB - Moderate support was obtained in a sample of 101 gay, bisexual, and heterosexual males for the perinatal hormone theory, which hypothesizes that attenuated levels of androgens during critical periods of male fetal development fail to masculinize and defeminize the brain. Affected individuals develop female-typical sexual orientation (assessed here by a pie chart) and cerebral organization, reflected in visual-spatial abilities and gender nonconformity. Handedness, also thought to reflect in utero hormone exposure, was evaluated. Gay and bisexual males reported greater femininity and lesser masculinity than heterosexuals, with bisexuals intermediate in masculinity, suggesting a common biological mediator for homoeroticism and sex atypicality. Among bisexual males, increased masculinity was related to enhanced performance on all spatial tasks. Group mean differences in spatial ability and handedness were not found; however, among bisexuals, poorer visual-spatial performance predicted increased homoeroticism and right-handedness positively correlated with all spatial tasks. If perinatal hormones contribute to a generalized feminization of the brain, the current data indicate that it is most apparent among bisexual males. Sexing of their brains may involve several sexually dimorphic regions that are related in a continuous manner. Inferred cerebral feminization was more circumscribed among gay and heterosexual males, for whom childhood sex atypicality was most highly distinguishing. Unspecified mechanisms responsible for homoeroticism in them may differ from those that produce same-sex attractions in bisexuals and thus have relatively little impact on other components of cerebral feminization. PMID- 11910787 TI - On the elusive nature of sex differences in cognition: hormonal influences contributing to within-sex variation. AB - We argue that within-sex variation resulting from the prenatal organizational and adult activational effects of gonadal steroid hormones has the potential to obscure between sex differences in cognitive performance and functional cerebral asymmetry. Two putative markers for prenatal testosterone, finger ridge count (FRC) asymmetry and the 2D:4D finger length ratio, have been linked to within-sex variation in cognitive performance. In particular, FRC allows the identification of men and women who show a reversal of the typical sex-related pattern of task performance. Three paradigms for the study of activational effects--seasonal, menstrual, and diurnal hormonal cycles--have evaluated changes in task performance and functional cerebral asymmetry. The performance of sex-dimorphic, but not sex-neutral, tasks changes with estrogen across the menstrual cycle and with testosterone across its seasonal and diurnal cycles. Functional cerebral asymmetry also changes systematically across both the menstrual cycle and the diurnal testosterone cycle in such a way that suggests left hemisphere performance increases as testosterone levels decline whereas right hemisphere performance increases as estrogen levels decline. In studies of sex differences, such correlates of within-sex hormone-related differences are rarely measured or controlled. Whatever the explanation for the associations of putative markers and hormone cycles with differences in cognitive abilities and cerebral asymmetry, it is clear that these relationships have the potential to contribute to the elusive nature of sex differences in cognition and functional brain organization. PMID- 11910788 TI - Sex differences in play fighting revisited: traditional and nontraditional mechanisms of sexual differentiation in rats. AB - In the traditional model for sexual differentiation in mammals, the female phenotype is the default condition. That is, the female-typical pattern will persist unless acted upon by hormones early in development. The frequency of play fighting in rats, as in most other mammals, is sexually differentiated, and conforms to the traditional model. Males engage in more play fighting than females, and this can be reduced to female-typical levels by neonatal castration. Furthermore, females can be induced to play fight at male-typical levels if treated with testosterone neonatally. Fractionation of play fighting into its constituent components, attack and defense, reveals that it is the frequency of attack that is sexually differentiated, not the likelihood of defense. However, in males, defensive behavior changes at puberty so that the play becomes "rougher." For males to switch to this rougher form of play fighting, they have to be androgenized perinatally. Hence, for males, this second aspect of play fighting that is sexually differentiated also follows the traditional model. In marked contrast, development of the female-typical pattern does not. Neonatal treatment of females with testosterone has no effect; at puberty, they still show the female-typical pattern. On the the other hand, ovariectomy, either at birth or at weaning, leads to females exhibiting the male-typical transition to rougher play fighting at puberty. That is, ovarian hormones appear to actively inhibit the expression of a male-typical trait in females. Play fighting, then, is a mixture of traits, with some features conforming to the traditional model and some not. For some phenotypic features, ovarian hormones appear to exert an active role in their development. PMID- 11910789 TI - Development of sexual partner preference in the zebra finch: a socially monogamous, pair-bonding animal. AB - Zebra finches are group living socially monogamous birds that pair for life. Partner preference is strongly sexually differentiated: males prefer to pair with females and females prefer to pair with males. Where do these preferences come from? What occurs during development that produces adult birds that pair with the opposite sex? There is surprisingly little animal research that addresses such questions, especially in species that form pair-bonds. Our research program focuses on two processes that may be involved in the development of adult sexual partner preference: (1) early (possibly organizational) hormone actions and (2) social experience. Females treated with estradiol or fadrozole (an estrogen synthesis inhibitor) as nestlings or embryos showed masculinized sexual-partner preference as adults, preferring to pair with other females even when potential male partners were available. Removal of adult males from breeding cages, so that young birds were not exposed to males or to male-female pairs during development, eliminated sex-typical partner preferences; these birds were equally interested in both sexes and were more likely than controls to pair with a same-sex partner. These experiments provide insights into the development of sexual-partner preference that may be applicable to other group living pair-bonding animals with biparental care. They also contribute to the foundation of animal research that is necessary for a biological approach to understanding the pair-bonding component of human sexual orientation. PMID- 11910790 TI - Prenatal exposure of the ovine fetus to androgens sexually differentiates the steroid feedback mechanisms that control gonadotropin releasing hormone secretion and disrupts ovarian cycles. AB - Exposure of the female sheep fetus to exogenous testosterone in early pregnancy permanently masculinizes the reproductive neuroendocrine axis. Specifically, in utero androgens given to female lambs from day 30 to 90 of a 147 day pregnancy dramatically altered the response of the gonadotropin releasing hormone (GnRH) neuronal network in the hypothalamus to both estrogen (E) and progesterone (P) feedback. Elevated concentrations of estrogen stimulated a massive release of GnRH in gonadectomized female sheep; however, male and androgenized female lambs were unable to respond to high E concentrations by producing this preovulatory like "surge" of GnRH. Further, the inhibitory actions of progesterone (P) were also sexually differentiated and adult males and androgenized females were much less responsive to P-negative feedback than normal ewes. The consequences of these abnormal steroid feedback mechanisms were reflected in the fact that only 72% of ovary-intact androgenized ewes exhibited normal estrous cycles in their first breeding season whereas none had a single estrous cycle during the second breeding season. In contrast, 100% of the control animals exhibited repeated reproductive cycles in both seasons. These data indicate that a relatively short exposure to male hormones during in utero life permanently alters the neural mechanisms that control reproduction and leads progressively to a state of infertility. PMID- 11910791 TI - Hormonal influences on sexual partner preference in rams. AB - Domestic rams display a naturally occurring variation in sexual partner preference, such that 6-10% of range-bred populations prefer male sexual partners (male-oriented) in contrast to the majority of rams that prefer female sexual partners (female-oriented). Male-oriented rams exhibit hormone profiles and stress responses distinctly different from their heterosexual counterparts. These differences include reduced circulating levels of testosterone that arise after anesthetization. Lower levels of aromatase activity in the medial preoptic area and estrogen receptor in the amygdala were also measured in male-oriented versus female-oriented rams and may represent an important link to sexual behavior that should be investigated. It is anticipated that the male-oriented ram model will be useful for studies aimed at identifying both the activational and organizational components and the neuronal substrates of male sexual partner preferences. PMID- 11910792 TI - Sexual partner preference in female Japanese macaques. AB - Whether animals ever exhibit a preference for same-sex sexual partners is a subject of debate. Japanese macaques represent excellent models for examining issues related to sexual preference in animals because females, in certain populations, routinely engage in both heterosexual and homosexual behavior over the course of their life spans. Multiple lines of evidence indicate that female homosexual behavior in Japanese macaques is a sexual behavior, not a sociosexual one. Additional evidence indicates that female Japanese macaques do not engage in homosexual behavior simply because acceptable male mates are unavailable or unmotivated to copulate. Patterns of sexual partner choice by female Japanese macaques that are the focus of intersexual competition indicate that females of this species choose same-sex sexual partners even when they are simultaneously presented with a motivated, opposite-sex alternative. Thus, in some populations of Japanese macaques, females prefer certain same-sex sexual partners relative to certain male mates, and vice versa. Taken together, this evidence suggests that female Japanese macaques are best characterized as bisexual in orientation, not preferentially homosexual or preferentially heterosexual. PMID- 11910793 TI - How many gay men owe their sexual orientation to fraternal birth order? AB - In men, sexual orientation correlates with the number of older brothers, each additional older brother increasing the odds of homosexuality by approximately 33%. However, this phenomenon, the fraternal birth order effect, accounts for the sexual orientation of only a proportion of gay men. To estimate the size of this proportion, we derived generalized forms of two epidemiological statistics, the attributable fraction and the population attributable fraction, which quantify the relationship between a condition and prior exposure to an agent that can cause it. In their common forms, these statistics are calculable only for 2 levels of exposure: exposed versus not-exposed. We developed a method applicable to agents with multiple levels of exposure--in this case, number of older brothers. This noniterative method, which requires the odds ratio from a prior logistic regression analysis, was then applied to a large contemporary sample of gay men. The results showed that roughly 1 gay man in 7 owes his sexual orientation to the fraternal birth order effect. They also showed that the effect of fraternal birth order would exceed all other causes of homosexuality in groups of gay men with 3 or more older brothers and would precisely equal all other causes in a theoretical group with 2.5 older brothers. Implications are suggested for the gay sib-pair linkage method of identifying genetic loci for homosexuality. PMID- 11910794 TI - Age of puberty and sexual orientation in a national probability sample. AB - The relations between sexual orientation and age of puberty in both men and women were examined in a national probability sample of the United States. The sample was the National Health and Social Life Survey (E. O. Laumann, J. H. Gagnon, R. T. Michael, & S. Michaels, 1994), which contains 3432 cases. Gay/bisexual men reported an earlier age of puberty (e.g., age of first pubic hair) relative to heterosexual men, but lesbian/bisexual women did not report a different age of puberty (i.e., age of menarche) than heterosexual women. These results confirm findings from prior research examining age of puberty using nonrepresentative samples and add to a body of literature suggesting that gay/bisexual men may score, on average, in the female-typical direction on certain sex-dimorphic physical and developmental characteristics. PMID- 11910795 TI - Gender-related traits of heterosexual and homosexual men and women. AB - Two studies investigated the relation between sexual orientation and gender related traits. Analyzing data from an Internet survey, Study 1 found that gay men and lesbians differed from same-sex heterosexuals most strongly on gender diagnosticity (GD) measures, which assess male- versus female-typicality of occupational preferences (effect sizes were 1.14 for men and 0.53 for women) and least strongly on instrumentality (I) and expressiveness (E). Study 2 found that GD measures showed large differences between 289 gay and 200 heterosexual men (d = 0.95) and between 296 lesbian and 435 heterosexual women (d = 1.32), whereas I and E showed much smaller differences. In Study 2 homosexual-heterosexual diagnosticity measures, computed from men's and women's occupational preferences, correlated very strongly with GD measures (r = 0.88 for men and 0.89 for women), indicating that occupational preference items that distinguished men from women also tended to distinguish heterosexual from homosexual individuals. LISREL 8 analyses showed that self-ascribed masculinity-femininity did not mediate the strong relation between sexual orientation and GD for men or for women. PMID- 11910796 TI - Hormonal mechanisms underlying aberrant sexual differentiation in male rats prenatally exposed to alcohol, stress, or both. AB - The male offspring of rats exposed to restraint stress, alcohol, or both during late pregnancy show normally masculinized genitalia; however, sexual differentiation of behavior is dissociated from the external morphology. In contrast to controls, males exposed prenatally to stress, alcohol, or a combination of these factors exhibited the female lordotic pattern. Thus, all 3 prenatal treatments led to incomplete behavioral defeminization. Behavioral masculinization was not altered by fetal alcohol exposure alone, but a significant number of males that experienced prenatal stress alone failed to copulate. A more severe disruption of behavioral masculinization occurred when stress and alcohol were combined. Very few males exposed to the combination treatment mated with females. This study attempted to relate the effects of these treatments on sexual behavior to the postparturitional surge in plasma testosterone (T) that is known to influence the process of sexual differentiation. Prenatally stressed males, like control males showed a large, brief surge in plasma T that peaked 1 hr after delivery. Altered defeminization and masculinization were seen in prenatally stressed males, despite a normal postparturitional T surge. Fetal alcohol exposure, with or without concomitant stress, depressed T to the same extent right after birth and led to a similarly blunted T surge 1 hr later. Thus, equal disruption of the neonatal T pattern occurred in alcohol-alone males, who showed normal male copulatory behavior, and in alcohol-plus-stress males, whose behavior was severely attenuated. The results suggest that consideration of abnormal exposure to T during prenatal ontogeny may be required to understand the atypical sexual behaviors associated with these treatments. PMID- 11910797 TI - Masculinization effects in the auditory system. AB - Otoacoustic emissions (OAEs) and auditory evoked potentials (AEPs) are different in several special populations of subjects. For females having opposite-sex co twins (OSDZ females) and for homosexual and bisexual females, OAEs are masculinized. Certain AEP measures from homosexual and bisexual females also are masculinized. Certain AEP measures from homosexual males are hypermasculinized. These and other facts can be explained by assuming that these special populations received greater-than-normal exposures to androgens at some point(s) during development, possibly during prenatal development. It is proposed that some differences in androgenization may have been spatially and temporally localized rather than global, and that the localized response to androgen exposure sometimes may be nonmonotonic. PMID- 11910798 TI - [Phylogenetic diversity of aerobic saprotrophic bacteria isolated from the Daqing oil field]. AB - A diverse and active microbial community in the stratal waters of the Daqing oil field (China), which is exploited with the use of water-flooding, was found to contain aerobic chemoheterotrophic bacteria (including hydrocarbon-oxidizing ones) and anaerobic fermentative, sulfate-reducing, and methanogenic bacteria. The aerobic bacteria were most abundant in the near-bottom zones of injection wells. Twenty pure cultures of aerobic saprotrophic bacteria were isolated from the stratal waters. Under laboratory conditions, they grew at temperatures, pH, and salinity values typical of the stratal water from which they were isolated. These isolates were found to be able to utilize crude oil and a wide range of hydrocarbons, fatty acids, and alcohols. Phylogenetic analysis carried out with the use of complete 16S rRNA sequences showed that the isolates could be divided into three major groups: gram-positive bacteria with a high and a low G + C content of DNA and gram-negative bacteria of the gamma-subclass of the Proteobacteria. Gram-positive isolates belonged to the genera Bacillus, Brevibacillus, Rhodococcus, Dietzia, Kocuria, Gordonia, Cellulomonas, and Clavibacter. Gram-negative isolates belonged to the genera Pseudomonas and Acinetobacter. In their 16S rRNA sequences, many isolates were similar to the known microbial species and some probably represented new species. PMID- 11910799 TI - [Intensity of the microbiological processes of the methane cycle in different types of Baltic lakes]. AB - The intensity of the microbiological processes of methane formation (MF) and methane oxidation (MO) was determined in the sediments and water of different types of Baltic lakes. The emission of methane from the lake sediments and methane distribution in the water column of the lakes were studied as functions of the lake productivity and hydrologic conditions. During summers, the intensity of MF in the lake sediments and waters varied from 0.001 to 106 ml CH4/(dm3 day) and from 0 to 3.2 ml CH4/(1 day), respectively, and the intensity of MO in the sediments and water varied from 0 to 11.2 ml CH4/(dm3 day) and from 0 to 1.1 ml CH4/(1 day), respectively. The total methane production (MP) in the lakes varied from 15 to 5000 ml CH4/(m2 day). In anoxic waters, the MP comprised 9-18% of the total PM in the lakes. The consumption of organic carbon for methanogenesis varied from 0.03 to 9.7 g/(m2 day). The role of the methane cycle in the degradation of organic matter in the lakes increased with their productivity. PMID- 11910800 TI - [Isolation of actinomycetes from soil using extremely high frequency radiation]. AB - A new method employing extremely high frequencies (EHFs) is proposed for the selective isolation of actinomycetes from soil. The pretreatment of soil suspensions with EHF wavelengths of 5.6 and 7.1 mm led to a nonselective isolation of actinomycetes. At the same time, the irradiation of soil suspensions within wavelength bands of 3.8-5.8 and 8-11.5 mm considerably augmented the total number of isolated actinomycetes and increased the fraction of the isolated rare genera by 2 and 7 times, respectively. The rare actinomycete genera were represented by Actinomadura, Microtetraspora, Nonomuraea, Micromonospora, Amycolatopsis, Pseudonocardia, Saccharotrix, and Streptosporangium. PMID- 11910801 TI - [The growth-promoting effect of Beijerinckia and Clostridium sp. cultures on some agricultural crops]. AB - New strains of Beijerinckia mobilis and Clostridium sp. isolated from the pea rhizosphere were studied with respect to their promoting effect on the growth and development of some agricultural crops. Seed soaking in bacterial suspensions followed by the soil application of the suspensions or their application by means of foliar spraying was found to be the most efficient method of bacterization. The application of B. mobilis and Clostridium sp. cultures in combination with mineral fertilizers increased the crop production by 1.5-2.5 times. The study of the population dynamics of B. mobilis by the method of genetic marking showed that this bacterium quickly colonized the rhizoplane of plants and, therefore, had characteristics of an r-strategist. At the same time, Clostridium sp. was closer to K-strategists, since this bacterium slowly colonized the econiches studied. The introduction of the bacteria into soil did not affect the indigenous soil bacterial complex. The presence of Clostridium sp. slowed down the colonization of roots by the fungal mycelium. The possible mechanisms of the plant growth-promoting activity of B. mobilis and Clostridium sp. are discussed. PMID- 11910802 TI - [New evidence for the ability of methylobacteria and methanotrophs to synthesize auxins]. PMID- 11910803 TI - [The role of bacterial growth autoregulators (alkyl hydroxybenzenes) in the response of staphylococci to stresses]. AB - The investigation of the response of a batch culture of Staphylococcus aureus to exogenous alkyl-substituted hydroxybenzenes (AHBs), chemical analogues of anabiosis autoinducers, showed that C1-AHB at concentrations from 5 microM to 1.5 mM did not influence the culture growth, whereas the more hydrophobic C6-AHB inhibited it at concentrations of 0.5 mM and higher. Either of the AHBs drastically enhanced the phenotypic dissociation of staphylococcal cultures, which manifested itself in an increase in the fraction of cells producing small nonhemolyzing colonies of G type when plated on solid media with erythrocytes. In a submerged staphylococcal culture, the relative number of cells producing G-type colonies varied from 10 to 90%, depending on the concentration of the AHB added. The growth of S. aureus in the presence of AHBs also enhanced cell tolerance to heat shock (heating at 45 or 60 degrees C for 10 min). The role of AHBs, which are structural modifiers of membranes and possess chaperone activity, in the mechanisms responsible for cell tolerance and phenotypic dissociation of microbial populations is discussed. PMID- 11910804 TI - [Elemental composition of extremely alkaliphilic anaerobic bacteria]. AB - The contents of several chemical elements were assessed in the haloalkaliphilic acetogenic bacterium Natroniella acetigena and the alkaliphilic sulfate-reducing bacterium Desulfonatronum lacustre using X-ray microanalysis, stereoscanning microscopy, and mass spectrometry. The organisms were found to differ significantly in their relative contents of S, K, P, and Cl. The P/S ratio in cells of the alkaliphilic bacteria studied grown on mineral media at different pH was pH-dependent. With a pH increase from 9 to 10, potassium extrusion from cells was observed, suggesting that secondary K+/H+ antiport activity accounts for the homeostasis of cytosolic pH. Deenergization of bacterial cells in the presence of inhibitors and ionophores results in specific changes in the P/S ratio, which may be considered an indicator of the cell energetic state. In Natroniella acetigena, the content of intracellular Cl was directly proportional to the NaCl concentration in the medium. Some metals were shown to be necessary for the N. acetigena viability; the requirement for Ni and Co was absolute. Although little demand for Mg was characteristic of the bacteria studied, their growth was stimulated by an increase in Mg concentration, and the cell resistance to lysis was enhanced. PMID- 11910805 TI - [Comparative study of the elemental composition of vegetative and dormant microbial cells]. AB - X-ray microanalysis showed that vegetative cells, viable resting forms, and nonviable forms (micromummies) of the bacteria Bacillus cereus and Micrococcus luteus and the yeast Saccharomyces cerevisiae differ in the contents of bioelements S, P, Ca, and K and the Ca/K and P/S ratios. Viable resting forms (cystlike refractory cells and bacillar endospores) had more calcium and less phosphorus and potassium than vegetative cells, the difference being higher for bacilli than for micrococci and yeasts. The distinctive feature of all viable resting microbial forms was their low P/S ratios and high Ca/K ratios. The differences revealed in the cellular content and ratios of bioelements probably reflect changes in ionic homeostasis accompanying the transition of vegetative microbial cells to the dormant state. Relevant potassium parameters indicate that the membranes of viable resting forms retain their barrier function. At the same time, the nonviable forms, even morphologically intact, of B. cereus and S. cerevisiae exhibited an anomalously low content of potassium, while those of M. luteus had an anomalously high content of this element. This suggests that the cellular membranes of micromummies lose their barrier function, which results in a free diffusion of potassium ions across the membranes. The possibility of using the elemental composition parameters for quick analysis of the physiological state of microorganisms in natural environments is discussed. PMID- 11910807 TI - [Microbial geochemical calcium cycle]. AB - The participation of microorganisms in the geochemical calcium cycle is the most important factor maintaining neutral conditions on the Earth. This cycle has profound influence on the fate of inorganic carbon, and, thereby, on the removal of CO2 from the atmosphere. The major part of calcium deposits was formed in the Precambrian, when prokaryotic biosphere predominated. After that, calcium recycling based on biogenic deposition by skeletal organisms became the main process. Among prokaryotes, only a few representatives, e.g., cyanobacteria, exhibit a special calcium function. The geochemical calcium cycle is made possible by the universal features of bacteria involved in biologically mediated reactions and is determined by the activities of microbial communities. In the prokaryotic system, the calcium cycle begins with the leaching of igneous rock predominantly through the action of the community of organotrophic organisms. The release of carbon dioxide to the soil air by organotrophic aerobes leads to leaching with carbonic acid and soda salinization. Under anoxic conditions, of major importance is the organic acid production by primary anaerobes (fermentative microorganisms). Calcium carbonate is precipitated by secondary anaerobes (sulfate reducers) and to a smaller degree by methanogens. The role of the cyanobacterial community in carbonate deposition is exposed by stromatolites, which are the most common organo-sedimentary Precambrian structures. Deposition of carbonates in cyanobacterial mats as a consequence of photoassimilation of CO2 does not appear to be a significant process. It is argued that carbonates were deposited at the boundary between the "soda continent", which emerged as a result of subaerial leaching with carbonic acid, and the ocean containing Ca2+. Such ecotones provided favorable conditions for the development of the benthic cyanobacterial community, which was a precursor of stromatolites. PMID- 11910806 TI - [Degradation of fucoidan by the marine proteobacterium Pseudoalteromonas citrea]. AB - It was found that Pseudoalteromonas citrea strains KMM 3296 and KMM 3298 isolated from the brown algae Fucus evanescens and Chorda filum, respectively, and strain 3297 isolated from the sea cucumber Apostichopus japonicus are able to degrade fucoidans. The fucoidanases of these strains efficiently degraded the fucoidan of brown algae at pH 6.5-7.0 and remained active at 40-50 degrees C. The endo-type hydrolysis of fucoidan resulted in the formation of sulfated alpha-L fucooligosaccharides. The other nine strains of P. citrea studied (including the type strain of this species), which were isolated from other habitats, were not able to degrade fucoidan. PMID- 11910808 TI - [Preparations of extracellular proteinases from Aspergillus ochraceus 513 and Aspergillus alliaceus 7dN1]. AB - Preparations of extracellular proteolytic enzymes with high anticoagulant activity resembling protein C activators were isolated from the culture liquids of Aspergillus ochraceus 513 and Aspergillus alliaceus 7 dN1 by precipitation with ammonium sulfate and subsequent purification from ammonium ions by gel filtration on a column with Sephadex G-25. The pH and temperature activity optima and stability of the proteolytic enzymes from A. ochraceus 513 and A. alliaceus 7 dN1 were determined. PMID- 11910809 TI - [Nonlinearity in the growth of bacterial colonies: conditions and causes]. AB - The universally recognized kinetic model of colony growth, introduced by Pirt, predicts a linear increase of colony size. The linearity follows from the assumption that the colony expands through the growth of only such cells that are located immediately behind the moving colony front, in the so-called peripheral zone of constant width and density. In this work, Pirt's model was tested on two bacteria--Alcaligenes sp. and Pseudomonas fluorescens--having markedly distinct cultural properties and grown on agarized medium with pyruvate. The colony size dynamics was followed for different densities of the inoculum, ranging from a single cell to a microdroplet of bacterial suspension (10(5)-10(6) cells), and for different depths of the agar layer, determining the amount of available substrate. A linear growth mode was observed only with P. fluorescens and only in the case of growth from a microdroplet. When originating from a single cell, colonies of both organisms displayed nonlinear growth with a distinct peak of Kr (the rate of colony radius increase) occurring after 2-3 days of growth. The growth of P. fluorescens colonies showed virtually no dependence on the depth of the agarized medium, whereas the rate of colony size increase of Alcaligenes sp. turned out to be directly related to the medium layer thickness. The departure from linearity is consistently explained by a new kinetic chart stipulating a possible contribution to the colony growth not only of peripheral cells but also (much more distinct in Alcaligenes) of cells at the colony center. The colony growth dynamics is determined not only by the concentration of the limiting substrate but also by the amount of autoinhibitor, the synthesis of which is governed by age of cells. The distinctions of growth from a single cell and microdroplet could also originate as a result of dissociation into the R- and S forms and competition between the corresponding subpopulations for oxygen and the common substrate. PMID- 11910810 TI - [The colony architectonics in Bacillus subtilis 2335]. AB - Colonies grown from vegetative B. subtilis 2335 cells had a standard structure, with bacillar cells occupying the whole colony volume. At the same time, the colonies of this bacterium grown from germinated spores had an abnormal structure characterized by the location of cells in a surface layer 100-200 microns thick at the colony boundary with the air. The glycocalyx of the colonies grown from spores was characterized by a wetting angle theta e of 120 degrees-160 degrees, whereas that of the colonies grown from vegetative cells had an angle theta e as low as 5 degrees-30 degrees. It is suggested that spores and vegetative cells follow different strategies of substrate colonization and that the architectonics of bacterial colonies is determined by the physicochemical properties of the glycocalyx. PMID- 11910811 TI - [Temperate bacteriophage ZF40 of Erwinia carotovora: phage particle structure and DNA restriction analysis]. AB - Structural organization of the temperate bacteriophage ZF40 of Erwinia carotovora was studied. Phage ZF40 proved to be a typical member of the Myoviridae family (morphotype A1). Phage particles consist of an isometric head 58.3 nm in diameter and a contractile 86.3-nm-long tail with a complex basal plate and short tail fibers (31.5 nm). Phage tail sheath, a truncated cone in shape, is characterized by specific packaging of structural subunits. The ZF40 phage genome is 45.8 kb in size, as determined by restriction analysis, and contains DNA cohesive ends. The ZF40 phage of Erwinia carotovora is assumed to be a new species of bacteriophages specific for enterobacteria. PMID- 11910812 TI - [Study of Erwinia carotovora phage resistance with the use of temperate bacteriophage ZF40]. AB - The causes of the unique phage resistance of the pectinolytic phytopathogenic strains of Erwinia carotovora were studied with the use of temperate bacteriophage ZF40. It was shown that, in these bacteria, the bacteriophage-cell interaction can be substantially blocked at the adsorption level. An adequate indicator for studying the temperate bacteriophages of erwinias was developed on the basis of mutants resistant to macromolecular bacteriocins. Various restriction-modification systems, which influence cell resistance to bacteriophages, were revealed for the first time in E. carotovora. The phage resistance was shown to be determined by the wide occurrence of homoimmune temperate viruses in pectinolytic erwinias. PMID- 11910813 TI - [Geochemical characteristics of the carbonate constructions formed during microbial oxidation of methane under anaerobic conditions]. AB - The aragonite constructions of the Black Sea are formed in a stable anaerobic zone and are a perfect object to study the natural mechanism of anaerobic methane oxidation. The most probable pathway of methane oxidation is its methanogen mediated reaction with bicarbonates, dissolved in seawater, with the formation of water and acetate, which is then consumed by other components of the anaerobic community. Comparison of the delta 13C values of carbonate minerals and organic matter once more demonstrated that the formation of the organic matter of biomass is accompanied by intense fractionation of carbon isotopes, as a result of which the total organic matter of biomass acquires an extremely light isotopic composition, characterized by delta 13C values as low as -83.8@1000. PMID- 11910814 TI - [Interview with Dr. W. Wiebicke, Bremen. To treat early with anti-inflammatory drugs!]. PMID- 11910815 TI - [Classics with innovative potential noted. The MMW drug prize for inhalative airway therapeutics]. PMID- 11910816 TI - [The patient has gastroesophageal reflux. Radio waves instead of antacids?]. PMID- 11910817 TI - [Sexual abuse--neglect--violence. Accordingly you diagnose child abuse]. PMID- 11910818 TI - [Phytotherapy. Advise your patients]. PMID- 11910819 TI - [Hepatitis C in Germany. More than 5000 new infections annually]. AB - The number of people infected with HCV in Germany is estimated to be 300,000. New infections total 5100 every year, with drug consumption playing a major transmission role. Since the introduction of routine tests, the risk of contracting HCV infection from donated blood has now decreased to less than 1 in 1 million. For diagnostic purposes, antibody screening assays, confirmation assays to exclude false positive results (immunoblot), core antigen assay and nucleic acid testing are employed. An effective vaccine against HCV infection is still not available, so that prevention of transmission must continue to be the main aim of prophylaxis. In this connection, every practicing physician is expected to know his infection status for HCV, HIV and HBV. PMID- 11910821 TI - [Use observations. On what studies can the physician rely?]. PMID- 11910820 TI - [Pegylated interferon plus ribavirin in chronic hepatitis C. More than half of patients show a lasting response]. AB - In the past ten years, treatment of chronic hepatitis C has made great advances. Only five years ago, not more than 16% of the patients experienced a lasting cure; now the HC virus can be eliminated in 54-56%. This has been made possible by an improvement to interferon-alpha by pegylation, that is, retardation, resulting in appreciably longer and more uniform active levels, together with improved eradication by the combination with ribavirin. At the same time, treatment has become more convenient for the patient, since interferon need now be administrated only once weekly, usually on an outpatient basis. In view of the side effects of interferon, treatment must be closely monitored. In the meantime, concrete treatment recommendations are available for every genetic type of HCV, for patients experiencing relapse, and for primary treatment failures. PMID- 11910822 TI - [Principles of general ultrasonographic practice. I. Basic concepts and definitions]. PMID- 11910824 TI - [Hormone replacement on new roads. Estrogens by nasal spray]. PMID- 11910823 TI - [The basic disease had already been diagnosed. Abdominal pain increased]. PMID- 11910825 TI - [A year after the introduction. Generic human insulin tested]. PMID- 11910826 TI - [Beta blockers for hypertension. No negative influence on insulin resistance]. PMID- 11910828 TI - [Geriatric assessment in 3 steps. Often mental decline is not correctly assessed]. PMID- 11910827 TI - [Antismoking pills for cardiac patients, too? Every third one rebounds]. PMID- 11910829 TI - [Bacterial airway infections. Standard antibiotics more often ineffective]. PMID- 11910830 TI - [Diabetic patient with hypertension takes A-I I blocker. Less likely dialysis]. PMID- 11910831 TI - [Official therapy guidelines for the family physician. Your time table in asthma]. PMID- 11910832 TI - [Continuation 29. Tachycardia: harmless or life threatening?]. PMID- 11910833 TI - [PARI: an acronym which is (also) an augury]. PMID- 11910834 TI - [PARI-ETLD (Therapeutic Approach and Nursing Research -- Epidemiology and Treatment of Decubitus Lesions]. AB - Pressure sores are one of the preferred nursing research topics but, in spite of the large number of studies, most questions related to the prevention and treatment of pressure sores remain unanswered. Well designed clinical trials and on sufficiently large samples are very rare and most treatments are routinely used even without a reliable evidence of their efficacy. The PARI-ETLD trial is the occasion for: a. evaluating the efficacy of the Fitostimoline, in the ri epitelization of superficial pressure sores; b. starting a clinical trial conducted by nurses; c. building a multicentre nursing network for collecting data on the epidemiology of pressure sores and for evaluating the effectiveness of caring strategies and treatments. The protocol presented, with the data collection forms, is an example of feasibility of clinical trials in the nursing practice and offers examples of ways for overcoming common problems related to the implementation of clinical trials in everyday practice. PMID- 11910835 TI - [Andrea's story]. AB - First-hand accounts of illness experiences provide important insights for other patients and their carers and can be a powerful tool for patient information and professional education. Andrea was ran over by a motor-bike while he was carried by bike and reported a complicated femur fracture. Three different representations of the story are reported and confronted: the bold chronicle of events, that sets the scenery and time sequence; Andrea's mother point of view on what happened after the accident, and during the course of the illness; and Andrea's story, told with his words and drawings. The methodological comments offered as discussion, stress how the collection of relevant patients stories can be a valuable research resource because it can offer a broad perspective which cannot be obtained by other means. PMID- 11910837 TI - [Assessment of the relationship between health personnel and caregivers caring for dementia patients. Results of a pilot survey]. AB - The characteristics of 129 patients admitted to 2 Nursing Homes (with and without Alzheimer ward) and one Geriatric Rehabilitation Unit and of their caregivers were described in this pilot study. Caregivers were asked to evaluate the quality of care received by the patients. Mental status of patients was assessed with the MM-SE, Functional status with ADL and behaviours with ICS and NPI. The stress of caregivers was evaluated with the Green's Relative Stress Scale. Patients with cognitive problems were admitted because of their illness (32.4%) or because impossible to care for them at home (20%), whilst for patients without cognitive problems difficulties in treatment at home (35.3%) and problems in walking (23.5%) represented the two main categories for admission. Caregivers are mostly women (65%) and the stress level of caregiver of patients with dementia is 22.7 (range 0-60) vs 14.2 of the other group. The overall judgement on the quality of care is good with few exceptions for the relationship with doctors and health personnel, and the level of competence of the health personnel of the Nursing homes without Alzheimer ward. The study was the occasion for a reflection on caregivers' expectations and wishes and for advancing proposals for a better answer to their needs. PMID- 11910836 TI - [Strategies to face the shortage of nurses in 11 health settings: descriptive analysis]. AB - A survey on the perceived severity and criteria for defining the nursing shortage was made by interviewing with a semistructured questionnaire 11 nurses responsible for nursing services of 11 Italian hospitals and districts. The questionnaire was administered by interview. Ten hospitals suffer a medium-severe nursing shortage. The main solutions adopted to overcome the nursing shortage are: the closure of wards, the merge of wards, with comparable patients; the employment of support personnel; the mobility of nursing personnel, whereby nurses change wards and are move where they are most needed; modification of shifts. Interestingly, criteria for defining the nursing shortage vary from hospital to hospital, but they are not based on objective data (c.g. number of "missing" nurses). The payment of overtime hours is also different, ranging from 20 to 35 dollars/hour. Since the shortage of nurses will last for the next 5-8 years, a study of criteria for assessing it, of solutions adopted and of their effectiveness is pivotal for gaining a better understanding of the problem. PMID- 11910838 TI - [Shift work at hospitals: comparison of organization models]. PMID- 11910839 TI - [What is efficacious for decubitus lesions]. PMID- 11910840 TI - [Violence against women. Report of Amnesty International]. PMID- 11910842 TI - Less invasive heart tests. PMID- 11910843 TI - I know breakfast is the most important meal of the day, but what are the elements of a good breakfast? PMID- 11910845 TI - Chronic pain: relief is out there. PMID- 11910844 TI - My hair is thinning on the top of my head. What treatments are available for hair loss in women? PMID- 11910847 TI - Aspirin: little pill, big potential. PMID- 11910846 TI - Advancing age is no defense against STDs. PMID- 11910848 TI - Too much vitamin A raises hip-fracture risk, except in women taking HRT. PMID- 11910849 TI - New dietary hints for kidney-stone prevention. PMID- 11910850 TI - Quality of life varies for breast cancer survivors. PMID- 11910851 TI - Arthritis. PMID- 11910852 TI - Drinking alcohol: don't start now. PMID- 11910853 TI - Hormone may be key to raloxifene's breast cancer benefits. PMID- 11910854 TI - Herbal remedies: are they safe and effective? PMID- 11910855 TI - Ditch the decaf? PMID- 11910856 TI - [Anthrax. The pathogen, the disease picture and possible use as a biological weapon]. PMID- 11910857 TI - [Autoantigens of subepidermal bullous autoimmune dermatoses]. AB - The dermal-epidermal junction contains a network of structural proteins that link epidermis and dermis. A central component of this complex is the cell membrane associated hemidesmosomal plaque. Formation of autoantibodies against different components of this hemidesmosomal anchoring complex can lead to subepidermal blisters. Such autoantibodies have been frequently used to characterize the target antigens at the molecular level. Autoimmune subepidermal blistering diseases include bullous pemphigoid, pemphigoid gestationis, lichen planus pemphigoides, linear IgA disease, cicatricial pemphigoid, anti-p450-, anti-p200- and anti-p105-pemphigoid, epidermolysis bullosa acquisita, bullous systemic lupus erythematosus and dermatitis herpetiformis Duhring. Differences in the clinical picture of these diseases can be attributed, at least in part, to the different specificity of the autoantibodies involved. The autoimmune response is further modulated by inflammatory cells and other inflammatory mediators. Native and recombinant forms of the autoantigens are increasingly used for the diagnosis of these diseases. PMID- 11910858 TI - [Fever as etiology of temporary infertility in the man]. AB - BACKGROUND AND OBJECTIVE: Fever of > or = 39 degrees C for > or = 3 days can lead to severely impaired semen quality and even azoospermia. Among spermatogenic cells, pachytene spermatocytes and spermatids are particularly sensitive to temperature. Because of the nature of a spermatogenic cycle, impairment of semen quality becomes obvious after a latent period of several weeks. The changes are usually reversible within a few months. Therefore, a careful history with regard to fever episodes during preceding 3 months is mandatory for interpretation of semen analysis. PATIENTS/METHODS: Five patients are reported who desired a child and had experienced high fever of > or = 39 degrees C for > or = 3 days. The results of semen analyses are presented in relation to fever episodes. RESULTS: Sperm concentration declined to 0.4-7% and fast progressive motility to 0-23% 3-6 weeks after the fever episodes and returned to base line levels within 4-6 months. In patients without semen analysis before the fever episode, semen quality remarkably improved thereafter. CONCLUSIONS: Investigating patients for a history of fever during the 3 months preceding fertility work-up avoids unreasonable treatment recommendations based on transient impairment of semen quality. PMID- 11910859 TI - ["Folie a deux" in the age of lasers]. AB - We report on a 59-year-old teacher suffering for approximately 5 years from multiple dermatological changes, some of which are artificially induced and some of which are imaginary. The patient believes these manifestations are the result of neighbors shooting at her with lasers. Her husband is also convinced of the veracity of these impressions. We interpret her notion of almost daily laser attacks to a skin-related paranoid/hallucinatory psychosis with the involvement of her husband in a "folie a deux". PMID- 11910860 TI - [Cutaneous manifestation of cysticercosis]. AB - Cysticercosis, an infection with the larva of Taenia solium, is caused by the accidental ingestion of the parasite's eggs. In many countries of the Third World, cysticercosis, and especially neurocysticercosis, is a widespread problem. A patient from Northern Malawi presented not only with cysticercosis but also with BT leprosy and pityriasis versicolor. Dermatologists should be familiar with the clinical picture of cysticercosis in order to make an early diagnosis in patients from at-risk areas. PMID- 11910861 TI - [Papular acantholytic dyskeratosis]. AB - Papular acantholytic dyskeratosis is a very rare skin disease, which shows a suprabasilar cleft with acantholytic and dyskeratotic cells, simulating the light microscopic picture of Darier disease. In this condition, the lesions are acquired without a genetic basis. We observed a 40-year-old woman with about 200 papules on the lower aspects of her legs; histological examination showed changes similar to Darier disease. Transmission electron microscopy showed a reduced number of desmosomes and perinuclear distribution of tonofilaments. The lesions were successfully treated with cryotherapy with spray technique. PMID- 11910862 TI - [Reproducible drug exanthema to terbinafine with characteristic distribution of baboon syndrome]. AB - A 26 year old patient developed a fixed drug eruption located on his hands, inguinal and gluteal areas following oral treatment of onychomycosis with terbinafine. The rash showed the characteristic distribution of the "baboon syndrome", so-named because of the red perianal region of the baboon. Although epicutaneous testing revealed no positive reaction, the rash could be induced in identical sites by oral administration of terbinafine. As the underlying pathomechanism for the "baboon-syndrome" a systemically induced allergic contact dermatitis has been suggested. In addition to the described substances, e.g. mercury, amoxicillin, ampicillin, heparin and nickel, this is the first report of "baboon syndrome" induced by terbinafine. PMID- 11910863 TI - [Muir-Torre syndrome]. AB - The Muir-Torre syndrome (MTS) is a rare, autosomal-dominant inherited disease characterized by sebaceous gland tumors and at least one internal malignancy. In many cases a genetic defect known as microsatellite instability can be identified. Similar pathogenetic mechanisms are found in patients with the hereditary non-polyposis colon cancer syndrome, so that at least some patients with MTS are considered as having a phenotypic variant of that syndrome. A 66 year-old woman with MTS, developed multiple malignant cutaneous and visceral tumors over 32 years; in addition, she had multiple sebaceous gland adenomas. Microsatellite instability could be proved with 2 out of 5 studied markers. The family history was positive as numerous relatives of the patient's mother were reported to have developed internal malignancies. PMID- 11910864 TI - [Post-zoster granuloma with detection of varicella zoster virus DNA in the granulomas]. AB - Granulomatous skin changes following herpes zoster are uncommon and their pathogenesis is unclear. We demonstrated varicella-zoster virus in the granuloma tissue of an immunocompromised patient with postherpetic granulomas and use this finding as basis for discussing the pathogenesis of these lesions. PMID- 11910865 TI - [What is your diagnosis? Verrucous plaque in a 3-year-old African child]. PMID- 11910866 TI - [Ethics in medicine. Needs and forms]. PMID- 11910867 TI - [Better tumor eradication in transgenic altered T-lymphocytes. Role of the TGF beta signal transduction cascade]. PMID- 11910868 TI - [Comment on W. Harth and R. Linse: "Botulinophilia"]. PMID- 11910869 TI - [Information for quality management in the hospital: on the function and methodology of patient and staph surveys]. AB - BACKGROUND: For quality management to be both systematic and realistic, a wealth of information is needed on the experiences patients as well as staff have in the hospital's daily running. This makes patient and staff surveys imperative to prudent quality management. TOPIC: The following article describes essential points for such studies to become methodologically sound and for their results to be of practical relevance. It is emphasized that the surveys' main purpose--i.e. to stimulate measures for improving the quality of care--can only be fulfilled when hospital staff, with the resolute support by its directors, is being successfully involved in these studies during all stages. PMID- 11910870 TI - [Training and educational measures in severe chronic heart failure. Experiences and application to general practice]. AB - OBJECTIVES: Prognosis and course of severe left ventricular dysfunction have been shown to be influenced positively by exercise training. Yet, physicians have been reluctant to include exercise into therapeutic concepts due to widespread uncertainty about the acceptable intensity of cardiac stress. Standardized exercise programs with proven safety may promote acceptance of this approach. PATIENTS AND METHODS: 25 patients with severe heart failure were enrolled in a PC controlled interval exercise training on cycle ergometer, lasting 21 minutes five times a week for a 4-week inpatient period. After discharge patients performed incremental daily walking for 3 months. Associated educational measures concerned life-style changing and self-controls. RESULTS: None of the patients had to interrupt the training because of side effects. 3 weeks and 3 months after starting exercise training there was an increase of peak VO2 from 13.4 +/- 2.8 to 14.5 +/- 3.8 (p < 0.05) and 15.3 +/- 3.2 ml/kg/min (p < 0.01), respectively, and an increase of ventilatory anaerobic threshold (VAT) from 9.3 +/- 1.9 to 10.0 +/- 2.3 (p < 0.05) and 11.3 +/- 2.2 ml/kg/min (p < 0.001), respectively. The improved aerobic capacity corresponded to a 9.3% increase in the results of the 6-minute walk test in the 4th week (n.s.). During out-patient period, the walking could be increased from 37 to 58 minutes daily. The physical quality of life was significantly improved after 3 months. There was no hospital admission due to heart failure. CONCLUSION: Exercise training in the therapy of severe heart failure is safe and increases the aerobic capacity. Associated educational measures may be able to minimize the danger of wrong exercise techniques and to reduce the rate of hospital readmissions. PMID- 11910871 TI - [Acute coronary syndrome--better risk stratification by determination of inflammatory parameters?]. AB - BACKGROUND: In the past years coronary atherosclerosis and plaque rupture have been characterized not only as a problem of growth of smooth vascular cells, of activated thrombocytes and coagulation but also of inflammation. PARAMETERS OF INFLAMMATION: Several studies have demonstrated the importance of C-reactive protein as a marker for cardiovascular complications. Also interleukin-6, tumor necrosis factor-alpha and adhesion molecules are discussed as predictors of a poor prognosis in patients with acute coronary syndromes. RISK STRATIFICATION: A positive troponin T test is correlated with an increased rate of early complications; thus, these patients benefit from an aggressive therapy with GP IIb/IIIa receptor antagonists and acute coronary intervention. The C-reactive protein is predictive for late complications such as restenosis, reinfarction and death due to cardiac events; these patients benefit from close controls and from an anti-inflammatory therapy with acetylsalicyclic acid and statins. PMID- 11910872 TI - [Measuring case severity with a DRG-based reimbursement system]. AB - BACKGROUND: The Australian Refined Diagnosis-Related Groups (AR-DRGs) will be the model for the German DRGs (G-DRGs). Their system to measure severity of illness will be a major point of interest. METHOD: The most common systems for measuring severity of illness are presented and compared with the AR-DRGs based on criteria regarding applicability. RESULTS: None of the systems for measuring severity of illness fits all the criteria. They can be used for reimbursement of inpatient care or for quality assurance, but not for both at the same time. The designated areas for the use of the systems should not be exceeded. CONCLUSION: AR-DRGs are very complex in measuring the costs per case (severity of illness in terms of efficiency). They are not able to support quality assessment by risk adjustment (severity of illness in terms of medical complexity). A less complex system would have been easier to transfer to Germany with the same incentives for providing effective care. PMID- 11910873 TI - [Primary prevention of cardiovascular diseases by nonpharmacological means]. AB - BACKGROUND AND AIM: Primary prevention with drugs such as ACE inhibitors, statins and betablockers is highly effective, but due to expensiveness not cost effective. The aim of this review article was to demonstrate whether nonpharmacological therapy with life-style factors (nonsmoking, healthy eating pattern, increased physical activity) can reduce the incidence of classic risk factors and the cardiovascular morbidity and mortality. METHODS: The effect of several important life-style factors such as different diets, nonsmoking and physical activity was analyzed by means of an inquiry of the literature. RESULTS: Randomized actual studies demonstrate that a healthy diet can influence the beginning and progression of hypertension and diabetes mellitus. Epidemiologic studies show with high consistency that nonsmoking, regular physical activity and an diet rich in fruit and vegetables, low-fat products, fish and lean meat is associated with a lower incidence of cardiovascular events and mortality. A further decrease of the cardiovascular morbidity can be reached by combining several of these life-style variables. CONCLUSIONS: Life-style factors such as a healthy eating pattern, nonsmoking and regular physical activity can contribute to an enormous health benefit in the general population and save money for the public social systems. Therefore the promotion of this healthy life-style should be a major aim of the health policy. PMID- 11910874 TI - [Fatal bleeding complications caused by Evans syndrome (autoimmune thrombocytopenia and hemolytic anemia) and type II autoimmune hepatitis in a 56 year-old patient]. AB - BACKGROUND: Autoimmune hepatitis is a rare form of hepatitis of nonviral origin. Two main subentities have been described. The classical lupoid hepatitis (type I) is characterized by hypergammaglobulinemia and the presence of lupus erythematosus cells due to antinuclear antibodies. Autoimmune hepatitis type II, which is associated with antiliver/kidney microsomal antibodies type 1 (LKM 1) shows a more aggressive clinical course than autoimmune hepatitis type I and is frequently (41% of cases) associated with other immunologic diseases. CASE REPORT: In the present study we report a case of autoimmune hepatitis Type II, associated with autoimmune thrombocytopenia and hemolytic anemia, in a 56-year old patient. The patient's death was caused by a fatal association of a failing coagulation system due to liver dysfunction and a severe autoimmune thrombocytopenia. The aggressive course of the thrombocytopenia even after splenectomy demonstrated that the splenic enlargement due to the portal hypertension was only a minor factor in the destruction of the thrombocytes. Interestingly, some findings of this case such as the advanced age, the presence of anti-smooth muscle antibodies and HLA-DR4 are usually associated with autoimmune hepatitis type I. CONCLUSION: The findings of this case indicate that concomitant autoimmune diseases can worsen the prognosis of autoimmune hepatitis. Prednisolone and azathioprine might not be sufficient to treat aggressive forms of autoimmune hepatitis. Immunosuppressive regimens administered to recipients after organ transplantation might be used as a therapy of autoimmune hepatitis in multicenter clinical trials. PMID- 11910875 TI - [Manifestation of Cushing syndrome and osteoporotic fractures in pregnancy in a patient with Carney complex]. AB - HISTORY AND CLINICAL FINDINGS: In a 31-year-old patient a conventional X-ray was performed due to persistent pain at the lumbar spine level after a cesarean section. It revealed compression fractures of L2 and L3. Besides very clear clinical signs of hypercortisolism, multiple hyperpigmentations and naevi in the patient's face including the lips and the conjunctiva of the right eye were visible, suggesting a Carney complex. INVESTIGATIONS: Insuppressible cortisol levels confirmed an adrenal origin of hypercortisolism. A selective catheterization of adrenal veins supported the presence of bilateral adrenal cortisol production. The computed tomography showed nodular lesions in the right and a hyperplasia of the left adrenal gland. TREATMENT AND COURSE: A bilateral adrenalectomy was performed and a primary pigmented nodular adrenal hyperplasia was confirmed histologically. Clinical signs of hypercortisolism rapidly resolved after adrenalectomy. CONCLUSIONS: The diagnosis of Cushing's syndrome as a part of Carney complex was diagnosed at the end of a pregnancy although signs of hypercortisolism were present a long time before. The rare diagnosis of Carney complex should be considered in patients exhibiting symptoms of hypercortisolim and the typical clinical signs (hyperpigmentations). PMID- 11910876 TI - [Acute renal failure as a sequela of mushroom poisoning with Cortinarius speciocissimus]. AB - BACKGROUND: Poisoning with the mushroom Cortinarius speciocissimus is a rare event. Within 2-17 days after ingestion unspecific symptoms appear. Characteristically, this state is soon followed by irreversible kidney damage which finally requires chronic hemodialysis. CASE REPORT: A 46-year-old man was admitted with nausea, vomiting and acute anuria 9 days after he had eaten Cortinarius speciocissimus. Hemodialysis had to be started. Kidney biopsy revealed an interstitial nephritis. This finding corresponds to earlier case reports about intoxication with orellanine, the poison of Cortinarius speciocissimus. 6 months later the kidney function had not recovered leaving the patient to chronic hemodialysis. CONCLUSION: The prognosis of poisoning with that mushroom is poor. The amount of ingested poison is crucial. Monitoring of kidney function is mandatory when poisoning is suspected. Therapy is directed to symptoms. Usually dialysis has to be commenced. To rule out other causes for kidney damage, a kidney biopsy is required. PMID- 11910878 TI - [Approach of menopause in women at risk for breast cancer]. AB - The small but significant increase in risk of discovering breast cancer in women with hormone replacement therapy and the recent discussion of coronary benefit of this treatment have led many authors to insist on the necessity to evaluate the benefit/risks ratio before administration. This evaluation is particularly important for women that are already at high risk of breast cancer because of some genetic predisposition, family history or some benign breast diseases. In these cases, it is important to evaluate the absolute risk of breast cancer, to define the patient's needs more precisely, to specify menopausal symptoms; it is also important to evaluate the risk of osteoporosis, to review the various therapeutic possibilities, which are not only estrogen/progestin treatments (there are alternative treatments), and to give the patients honest information. Before obtaining the results of current trials, we are proposing here a pragmatic attitude and a decision algorithm to adopt a therapeutic attitude more easily which will be decided together by both patients and their physicians. PMID- 11910877 TI - [Metformin-associated lactic acidosis with acute renal failure in type 2 diabetes mellitus]. AB - CASE REPORT: An 83-year-old patient was admitted to our hospital because of gastrointestinal symptoms, mental confusion and dysarthria. The patient suffered from type 2 diabetes mellitus and was taking metformin. A mild renal insufficiency was known. On admission, we found impaired consciousness, Kussmaul breathing, a body temperature of 32.1 degrees C, and hemodynamic instability. Laboratory testing revealed lactic acidosis (pH 6.71, base excess--30, standard bicarbonate 4.0 mmol/l, lactate 24.4 mmol/l) and acute renal failure with a creatinine of 10.6 mg/dl and blood urea nitrogen of 134 mg/dl. Electrolytes were not altered; the blood glucose was elevated (147 mg/dl). According to history, physical examination, and laboratory testing the diagnosis metformin-induced lactic acidosis with acute renal failure was made. This diagnosis was confirmed by an elevated level of metformin. As soon as possible a bicarbonate hemodialysis was initiated. After 8 hours of hemodialysis the acid-base metabolism was almost balanced and the vigilance of the patient normalized. No further sessions of hemodialysis were needed and insulin therapy was started. CONCLUSIONS: Metformin induced lactic acidosis is a common side effect in patients with renal insufficiency. For an early diagnosis, clinical symptoms of intoxication should be well known by physicians and patients. First-line therapy for correction of lactic acidosis and effective elimination of metformin is bicarbonate hemodialysis. Sodium bicarbonate infusions are not able to correct the acid-base metabolism sufficiently. For prevention the renal function should be monitored closely and metaformin therapy should be stopped, if a deterioration of renal function is observed. PMID- 11910879 TI - [Laparoscopic treatment of genital prolapse: lateral utero-vaginal suspension with 2 meshes. Results of a series of 47 patients]. AB - We report our experience with a new technique to treat genital prolapse: the laparoscopic lateral suspension with two meshes. This is a prospective longitudinal study of 47 women with genital prolapse. With a mean follow-up of 15.2 +/- 10 months (1-39), the anatomical result was perfect in 78.7% of the cases (37 patients). From the functional point of view, 89.3% were satisfied (42 patients). These good preliminary results need to be confirmed by other extensive studies. PMID- 11910881 TI - [Is the limitation to 6 cycles of insemination with donor sperm justified?]. AB - OBJECTIVE: To evaluate the effect of the limitation at 6 cycles of artificial insemination in a program of sperm donation. STUDY DESIGN: 266 couples included in a program of sperm donation underwent 1,354 cycles including 532 intracervical insemination (ICI), 678 intrauterine insemination (IUI) and 133 in vitro fertilization (IVF). RESULTS: The birth rate by cycle was 10.8% in ICI, 18% in IUI, 21.9% in IVF. The risk of multiple pregnancies was 0% in ICI, 13% in IUI, 33% in IVF. The authors feign then two strategies, the first one with 6 ICI followed by 6 IUI, and the other one with 6 IUI alone. The birth rate, the risk of multiple pregnancies and the cost of these two strategies is discussed. CONCLUSION: The restriction to 6 IA Cycles in a donor semen program does not change the birth rate, but increases the multiple pregnancy rate and the cost of the treatment of these patients. PMID- 11910880 TI - [Evolution of homocysteine during ovarian stimulation for IVF or ICSI]. AB - OBJECTIVE: To determine the homocysteine evolution during ovarian stimulation in IVF or ICSI protocols and in, a second time, to evaluate the role of hyperhomocysteine as thrombotic risk factor for the treated patients. MATERIAL AND METHODS: Plasma homocysteine was determined three times for each of 31 women included in an IVF/ICSI program. Dosages were realised before stimulation, after gonadotrophin-releasing hormone agonist treatment (GnRH) and on the day of hCG injection. Vitamin B12 and folates were determined before stimulation. In case of hyperhomocysteinemia, a research of APCR (Activated Protein C Resistance) was realised. RESULTS: Five hyperhomocysteinemia cases were discovered (16.12% of studied population). APCR was found in a patient with hyperhomocysteinemia (14 mumol/L, before stimulation). Molecular biology has confirmed an heterozygous mutation of factor V Leiden. During the ovarian stimulation the evolution of homocysteine was independent of the 17 beta oestradiol evolution. CONCLUSION: The prevalence of hyperhomocysteinemia was not significative according to the limited size of the studied population. The increase of oestradiol during induction protocols is unrelated to the homocysteine level. This work must be continued with largest population to have better knowledge of the prevalence of hyperhomocysteinemia among women included in ovarian stimulation protocols. PMID- 11910882 TI - [Hemoperitoneum secondary to the rupture of superficial veins of a uterine leiomyoma]. AB - The most frequent cause of gynaecological hemoperitoneum is ruptured ectopic pregnancy. An uncommon cause of hemoperitoneum is rupture of uterine leiomyoma. We report one case of massive intraperitoneal hemorrhage and hypovolemic shock due to ruptured uterine leiomyoma vessels. The patient was treated with myomectomy and the postoperative course was uneventful. These case suggest that massive intra peritoneal hemorrhage associated with uterine leiomyoma should be considered in women with hypovolemic shock and pelvic mass. PMID- 11910883 TI - [Sexual fantasies and impotence. Apropos of 2 cases]. AB - Sexual fantasies, drawn of the erotic memory that constitutes himself the long of the sexual history of each, are indispensable, for the good sexual working. Some authors tend to consider the imagination like a "veritable intra psychic erogenous zone". However, people who, for a reason or an another, persisted to avoid all sexual activity before the marriage and to separate all erotic velleity, don't they risk to have an erotic imaginary atrophied, who could sound negatively once on their sexual behaviour, after the marriage, when all is suddenly authorized for them? Through two clinical observations of secondary sexual impotence, we are going to try to show the ominous consequences of such a relentlessness (imposed by the education, the social morals or the religion), against sexual stimulation, on the sexuality. These consequences are more easily observable in men, of the fact of their active role, in the sexual activity. Our purpose is to insist on the necessity to explore the sexual history owing all sexual dysfunction, and to place it in the biographic setting of the impatient; but also and especially to attract the attention on the importance of a precocious sex education, adapted to age, to the personal maturation and the cultural and educational context. PMID- 11910884 TI - [Epidemiology and natural history of genital infection by human papillomavirus]. AB - Human papillomavirus genital infection is a very common sexual transmitted disease, probably the most common of them. On one hand, this infection is more often than not transient and asymptomatic and induces an effective immunity which allows the infection cure; on the other hand it can be responsible for an intraepithelial lesion which can progress to an invasive cancer. In spite of the decrease of cervical cancer incidence thanks to Pap smear screening, it remains a real preoccupation for clinicians. If HPV is not sufficient for cervical carcinogenesis, it represents however a necessary factor. Near 100% of cervical cancers are indeed positive in HPV DNA. HPV infection is very frequent in young people aged less than 25 years and viral clearance average is 8 months. This clearance is the consequence of host immunity intervention which leads to spontaneous regression of infection and of the overwhelming majority of low grade squamous intraepithelial lesions (more than 80% within a period of two years). The major factor which permits the progression to high grade squamous intraepithelial lesions is the persistent feature of HPV infection. Cervical cancer is clearly the first viral-induced solid tumor discovered in human species. Furthermore it represents a woman death cause that can be avoided. PMID- 11910885 TI - [Contribution of high resolution breast ultrasonography in the characterization of ambiguous mammograms]. AB - The near-field imaging capability of sonography equipment has recently and markedly improved. High-frequency ultrasonography can improve the specificity of clinically and mammographically detected abnormalities, and helps accurately distinguish benign solid nodules from indeterminate or malignant nodules. The aim of this distinction is to obviate surgical biopsy. PMID- 11910886 TI - [3rd generation progestagen and thromboembolic disease. How do we decode the information?]. AB - In 1995-1996, four studies published in different medical papers showed an increased risk of veinous thromboembolism among women using third-generation contraceptives compared with those using second-generation pills. These papers had an important mediatic impact. Nevertheless, rereading of the scientific information showed important bias in these studies (methodology, prescription, diagnosis). More recently, two studies published in the British Medical Journal analysing the venous thromboembolism (VTE) relative risk with the third generation contraceptives and using the same database (General Practitioner Research Database) found contradictory results, introducing some confusion. The use of a third-generation progestagen does seem to have led to an unexpected and yet unexplained return of a higher risk of venous thromboembolism but the risk has certainly been overestimated in the first studies. PMID- 11910887 TI - [Prolongation of embryonic cultures and value for blastocyst stage transfer. There is a benefit to prolonging embryonic cultures to the blastocyst stage]. PMID- 11910888 TI - [Prolongation of embryonic cultures and value for blastocyst stage transfer. There is no benefit to prolonging embryonic cultures to the blastocyst stage]. PMID- 11910889 TI - [In response to the article by P. Faucher et al. Management of pregnant women infected by HIV at Bichat hospital between 1990 and 1998: analysis of 202 pregnancies. Gynecol Obstet Fertil 2001; 29: 211-5]. PMID- 11910890 TI - [In response to the article by A. Agostini et al. Significance of pelvic packing in persistent hemorrhage after hysterectomy for hemostasis. Gynecol Obstet Fertil 2001: 29: 613-5]. PMID- 11910891 TI - [Our responsibility to children of assisted reproduction ]. PMID- 11910892 TI - Chemokine receptors: multifaceted therapeutic targets. AB - Chemokines and their receptors are involved in the pathogenesis of diseases ranging from asthma to AIDS. Chemokine receptors are G-protein-coupled serpentine receptors that present attractive tractable targets for the pharmaceutical industry. It is only ten years since the first chemokine receptor was discovered, and the rapidly expanding number of antagonists holds promise for new medicines to combat diseases that are currently incurable. Here, I focus on the rationale for developing antagonists of chemokine receptors for inflammatory disorders and AIDS, and the accumulating evidence that favours this strategy despite the apparent redundancy in the chemokine system. PMID- 11910893 TI - The B7-CD28 superfamily. AB - The B7-1/B7-2-CD28/CTLA-4 pathway is crucial in regulating T-cell activation and tolerance. New B7 and CD28 molecules have recently been discovered and new pathways have been delineated that seem to be important for regulating the responses of previously activated T cells. Several B7 homologues are expressed on cells other than professional antigen-presenting cells, indicating new mechanisms for regulating T-cell responses in peripheral tissues. Some B7 homologues have unknown receptors, indicating that other immunoregulatory pathways remain to be described. Here, we summarize our current understanding of the new members of the B7 and CD28 families, and discuss their therapeutic potential. PMID- 11910894 TI - Redefinition of lymphoid progenitors. AB - Similarities between T and B lymphocytes might have led to the idea that these functionally distinct cells develop from a common lymphoid progenitor. However, investigations with a new clonal assay which allows for T-, B- and myeloid lineage development indicate that commitment to T-cell and B-cell lineages occurs instead through myeloid/T and myeloid/B bipotential stages, respectively. These findings provide an opportunity to reconsider the ontogeny and phylogeny of T- and B-cell development. PMID- 11910895 TI - Relationship of viral infections to wheezing illnesses and asthma. AB - Viral infections can influence both the development and the severity of asthma. In early life, viral infections can either increase or, remarkably, decrease the risk of subsequent asthma. In children and adults with existing asthma, viral respiratory infections frequently cause acute airway obstruction and wheezing. This article discusses the influence of viral infections on mechanisms of virus induced airway inflammation in relationship to the development, persistence and severity of asthma. PMID- 11910896 TI - MIF keeps macrophages on guard. PMID- 11910897 TI - Making a commitment. PMID- 11910898 TI - C-type lectin receptors on dendritic cells and Langerhans cells. AB - Dendritic cells and Langerhans cells are specialized for the recognition of pathogens and have a pivotal role in the control of immunity. As guardians of the immune system, they are present in essentially every organ and tissue, where they operate at the interface of innate and acquired immunity. Recently, several C type lectin and lectin-like receptors have been characterized that are expressed abundantly on the surface of these professional antigen-presenting cells. It is now becoming clear that lectin receptors not only serve as antigen receptors but also regulate the migration of dendritic cells and their interaction with lymphocytes. PMID- 11910899 TI - Epitope spreading in immune-mediated diseases: implications for immunotherapy. AB - Evidence continues to accumulate supporting the hypothesis that tissue damage during an immune response can lead to the priming of self-reactive T and/or B lymphocytes, regardless of the specificity of the initial insult. This review will focus primarily on epitope spreading at the T-cell level. Understanding the cellular and molecular basis of epitope spreading in various chronic immune mediated human diseases and their animal models is crucial to understanding the pathogenesis of these diseases and to the ultimate goal of designing antigen specific treatments. PMID- 11910901 TI - Journey towards excellence. PMID- 11910900 TI - Lipid rafts and B-cell activation. AB - The B-cell antigen receptor acts during B-cell activation both to initiate signalling cascades and to transport antigen into the cell for subsequent processing and presentation. Recent evidence indicates that membrane microdomains, termed lipid rafts, have a role in B-cell activation as platforms for B-cell receptor (BCR) signalling and might also act in antigen trafficking. Lipid rafts might facilitate the regulation of the BCR during B-cell development by B-cell co-receptors, and during viral infection. So, lipid rafts seem to be an important new piece of the B-cell signalling puzzle. PMID- 11910902 TI - Thumbsucking can affect a child's smile. PMID- 11910903 TI - Dentists can be better informed about child abuse issues. PMID- 11910904 TI - A resolution we can all share. PMID- 11910905 TI - Hawaii's first lawsuit alleging AIDS discrimination. PMID- 11910906 TI - "Am I legally obligated to provide dental care to an HIV infected patient?". PMID- 11910907 TI - AIDS dental discrimination case settled. PMID- 11910908 TI - A season of hope. PMID- 11910909 TI - Winning the battle against overhead. PMID- 11910910 TI - Trade imbalance or brain imbalance? PMID- 11910911 TI - "Da way I see it". PMID- 11910913 TI - New artificial tooth more accurate for teaching dental students. PMID- 11910912 TI - Disagreement with HMSA and with the purpose of the ADA's Code. PMID- 11910914 TI - JAWS-3D records jaw movements. PMID- 11910915 TI - We are growing. PMID- 11910916 TI - TB is on the rise. PMID- 11910917 TI - Approach change with cautious eyes. PMID- 11910919 TI - Hawaii Medical Service Association is no longer reimbursing patients under code number 3110 Pulp cap-direct or 3120 Pulp cap-indirect of the American Dental Association's Code on Dental Procedure and Nomenclature. PMID- 11910918 TI - More complicated than just a coding issue. PMID- 11910920 TI - Non-Hodgkins lymphoma of the oral cavity associated with HIV infection. PMID- 11910921 TI - An immediate and enormous threat to dentistry. PMID- 11910922 TI - Fairy ridiculous. PMID- 11910924 TI - HIV supplemental income protection when accidentally exposed. PMID- 11910923 TI - Behavioral pediatric dentistry, human communication, and Hawaii. PMID- 11910925 TI - The propriety and wisdom of obligating the HDA to a debt of 3.8 million dollars for 30 years there are a few additional points to be made. PMID- 11910926 TI - Hawaii Dental Service. PMID- 11910927 TI - Maui Medicaid clinic. PMID- 11910928 TI - Maui Dental Coalition. PMID- 11910929 TI - How do you clean teeth in your office? PMID- 11910930 TI - Dentistry in China: revelations of a western visitor. PMID- 11910931 TI - Effective chairside manner. Part 2. PMID- 11910932 TI - Mind quake. PMID- 11910933 TI - [Usefulness of immunohistochemistry as a diagnostic tool for tumors and pseudotumoral bone lesions]. AB - Immunohistochemical study for the diagnosis of bone tumors and tumor-like lesions has to be scheduled after an appropriate analysis of clinical data, radiological findings, and results of histology in H-E sections. The value of several markers for osteoblasts is discussed, chiefly for various forms of osteosarcomas. In the same way, the role of S-100 protein as well as anticollagen type II antibody is developed for cartilaginous tumors. The selection of markers in the fields of round cell tumors and spindle cell tumors of bone is also discussed. Some diagnostic problems with the support of immunohistochemistry are described, like chordomas versus chondrosarcomas or bone metastases. Lastly, immunohistochemical study of proliferating factors in the bone tumor field is quoted. PMID- 11910934 TI - [Chondroblastoma and its differential diagnosis]. AB - Chondroblastoma is one of the most well defined cartilaginous tumors of bone on imaging and pathological findings. However in practice, the pathologist is faced with diagnostic difficulties. The most recent features of the tumor are developed in the review: clinical findings, skeletal distribution, and above all, the results of the imaging study which has to be strictly correlated with histology for the diagnosis as well as for the differential diagnosis, location, osteolysis, matrix production, periosteal reactions, MRI studies. Histopathology of chondroblastoma is described: cellular component, architecture, and secondary changes. Usefulness of cytopathology is also stressed on smears or tumor imprints. Main findings of recent and various immunohistochemical stains are described and discussed as well. Ultrastructural and cytogenetic studies are summarized. Treatment, natural history, and prognosis of the tumor are developed. The three most important differential diagnoses are giant cell tumors, clear cell chondrosarcomas, and aneurysmal bone cysts. PMID- 11910935 TI - [Embryology and pathology of the human notochord]. AB - The scope of this review is the report of recent findings on the developmental anatomy of the human notochord and vertebral column and the related tumors, that is chordomas. Anatomy and histology of adult intervertebral discs are also described. All the topics are completed by histochemical and immunohistochemical findings. Location and incidence of notochordal vestiges in the adult are correlated to the development of chordomas which are also described for their clinical features, skeletal distribution, spread, prognosis, and treatment. Pathology of chordomas includes morbid anatomy, histopathology, cytology, and ultrastructural findings. The seemingly distinct group of chondroid chordomas is also described. The link between chordomas and notochordal cells is stressed. PMID- 11910936 TI - [Cellular and molecular biology of fibrous dysplasia]. AB - Several recent studies reflect major progress on the molecular biology and on the mechanisms of the lesions of fibrous dysplasia (monostotic and polyostotic forms) as well as the McCune-Albright syndrome. This review includes a detailed histological description of fibrous dysplasia: disorganized collagen fibers, woven bone formation, immature cytology of the osteoblasts and preosteoblastic cells. Histological lesions are also associated with an alteration of bone proteins: such as an increase in the expression of osteonectin, decrease of osteopontin and bone sialoprotein. Cell cultures of dysplasic osteoblastic cells have shown that the bone lesions are the result of abnormalities of the proliferation and differentiation of bone forming cells with a rise in CAMP levels. Poorly differentiated cells are associated with the formation of disorganized collagen fibers and with a trouble of synthesis of bone proteins. All these findings are linked to mutations of the Gs alpha (sub-unit alpha) protein. Various activating mutations are described as well as the molecular mechanisms of different forms of fibrous dysplasia, involving specially the c-fos proto-oncogene. Therapeutic implications are quoted, specially calcitonin and bisphosphonates. PMID- 11910937 TI - [Adamantinoma of the long bones: an anatomo-clinical review and its relationship with osteofibrous dysplasia]. AB - Adamantinoma of long bones is a perplexing tumor for its histology as well as for its histogenesis. Recent progress has been made with ultrastructural and immunohistochemical works. The possible relationship with osteofibrous dysplasia is the subject of conflicting discussions and the potential link has implications for the diagnosis, prognosis, and treatment. Clinical aspects of adamantinomas are described: location, age, gender, symptoms, imaging, diagnosis, treatment, clinical course, prognosis. Histopathology is extensively covered for usual and peculiar forms. Immunohistochemical results are described and discussed. Data of electron microscopy, cytological findings, and DNA flow cytometry are also reported. The putative association with osteofibrous dysplasia is the subject of an extensive development with the report of the personal experience and works of the authors, and arguments for viewing osteofibrous dysplasia and "osteofibrous dysplasia-like" adamantinomas as precursor lesions of classic adamantinomas are favoured. PMID- 11910938 TI - [Osteonecrosis]. AB - In orthopedic pathology, the pathologist is most commonly faced with the study of resected femoral heads for osteonecrosis. Such a study necessitates a knowledge of clinical findings, of physiopathology and chiefly of radiological findings. Osteonecrosis of the femoral head following fracture has been reported with four morphological stages which are very precisely described. In the same way, idiopathic, non traumatic, or primary osteonecrosis is covered, stressing the putative etiological factors and the most important findings of imaging. The review includes also the skeletal manifestations of decompression sickness as well as bone infarctions not associated with caisson disease, and spontaneous osteonecrosis of the knee. From the recruitment of the Hospital for Special Surgery, NY, it has been recently reported that a significant number of patients regarded as cases of osteonecrosis, either in the femoral head or in the knee, are actually cases of subchondral insufficiency fractures. PMID- 11910939 TI - [Membranous lipodystrophy of the bone]. AB - Membranous lipodystrophy is a rare disease in which cyst-like lesions of fat occur in subcutaneous and other sites including bone marrow, together with sudanophilic leukoencephalopathy. The disease has been reported mostly in Finland and in Japan, with sporadic cases from USA, Belgium, Italy, and France. The cyst like lesions of limb bones progress in early adult life, usually followed by neurological disorders including convulsions and presenile dementia. Histologically, the fat cells of the bone marrow, synovial membrane, and other sites, are replaced by a convoluted, hyaline, eosinophilic membrane that surrounds a large space. The histochemical and ultrastructural characteristics of the membranes are described. In the brain, atrophy of the subcortical white matter of the frontal and temporal lobes with marked astrocytosis and fibrillary gliosis, and a slight or moderate degeneration of myelin sheaths, are the most prominent changes. Genetic studies of the disease have been reported, revealing an autosomal recessive gene, explaining both its sporadic and familial occurrence, with mutations involving the gene encoding a transmembrane protein, key activating signal transduction element in NK cells (TYROBP). The pathogenesis of membranocystic changes is still unknown: result of a metabolic disorder or circulatory disturbances. PMID- 11910940 TI - [Erdheim-Chester disease. Apropos of a case with autopsy findings]. AB - Erdheim-Chester's Disease is a very uncommon variety of non-Langerhans histiocytosis of unknown etiology, which characteristically affects long bones bilaterally and symmetrically in adults. It may be accompanied by visceral foci of variable localization and extension determining prognosis. Bone scintigraphy is characteristic enough to evoke the disease but histologic examination of a peripheral specimen is required to confirm the diagnosis: spumous histiocytes CD68+, PS100+/-, CD1a-. We describe a case revealed by a severe lung disease with detailed autopsy. PMID- 11910941 TI - [A subcutaneous myxoid nodule of the shoulder]. PMID- 11910943 TI - The OIIQ assesses the nursing extern program before the press. PMID- 11910942 TI - [A subcutaneous nodule of the ring finger]. PMID- 11910945 TI - Nurses and social workers called upon to work as partners. PMID- 11910947 TI - Fear, shame, and health promotion. PMID- 11910948 TI - Social stigma and negative consequences: factors that influence college students' decisions to seek testing for sexually transmitted infections. AB - College students often delay or avoid seeking testing for sexually transmitted infections (STIs), even if the services are readily available. We used in-depth, semistructured interviews to survey 41 college students aged 18 to 23 years about factors that influence decisions about STI testing. We grouped statements into 9 themes that represent influences on the decision. The most frequently mentioned factors were negative consequences of testing and perceived vulnerability to infection; other issues that influenced decision making included perceived benefits, perceived severity of diseases, public knowledge and opinion, social norms, provider characteristics, test-site characteristics, and personal considerations. Social stigmas and negative consequences appear to represent significant barriers to college students' being tested, which could increase the risk of spreading infections to others. Clinicians and health educators should raise students' awareness of the need for screening and should work to reduce the barriers to screening, including social stigmas and negative consequences. PMID- 11910949 TI - Trauma screening in students attending a medical university. AB - Clinicians seldom assess trauma history in patients who seek treatment for psychological problems, yet trauma exposure is often related to psychological distress. Assessing trauma history can provide valuable information for treatment conceptualization and provision, although patients may not spontaneously share their histories because of embarrassment, avoidance, or other concerns. The authors compared 73 students at a southeastern US medical university who sought counseling and psychological services and completed intake paperwork without a trauma screen with 130 students whose intake procedures included trauma screening. They found that (a) patients who were specifically asked about trauma history were more likely to report such events, (b) previous physical assaults with a weapon were related to current psychological distress, and (c) physical assaults with or without a weapon were related to clinically significant psychological distress. These findings suggest that screening for lifetime trauma history should be a standard part of mental health screenings in similar medical university counseling centers. PMID- 11910950 TI - Freshman 15: valid theory or harmful myth? AB - The authors investigated whether the perception that freshmen gain 15 pounds during their 1st year of college is related to either actual or perceived weight gain. Forty-nine incoming freshmen at a small liberal arts college completed the study by filling out questionnaires and health data at the beginning and end of their 1st year on campus. The findings revealed no significant weight gain at the end of the year. The "Freshman 15" myth was found to play an important role in perpetuating negative attitudes toward weight. Freshmen who were concerned about gaining 15 pounds were more likely to think about their weight, have a poorer body image than others, and categorize themselves as being overweight. PMID- 11910952 TI - Using a chain-of-effects framework to meet institutional accountability demands. PMID- 11910951 TI - Vaginal douching practices among women attending a university in the southern United States. AB - The authors assessed the frequency, characteristics, and motivational antecedents of vaginal douching practices among 125 White and 155 Black female college students. Overall, 40% of the students had ever douched and half of those women currently douche. Black women were most likely to be encouraged to douche by their mothers, whereas White women were more influenced by television advertisements. Among the sexually active women, being Black, using oral contraceptives, using spermicides, and being encouraged to douche by their mothers or by the media were independently associated with ever having douched. These associations were present among both Black and White women and were stronger when current douching was compared with never having douched. Women who were discouraged from douching by a physician or nurse were more likely to have stopped the practice. Douching is common, even among educated young women; nurses' and physicians' advice to stop douching appears to have a salutary effect. PMID- 11910953 TI - A cross-sectional audit of student health insurance waiver forms: an assessment of reliability and compliance. AB - To assess the reliability of using a waiver process to ensure compliance with health insurance requirements established by a university, the author conducted a cross-sectional verification and compliance audit of insurance waiver forms received for the 1999/2000 academic year. This study revealed that a waiver form process could not be relied upon to enforce compliance. PMID- 11910954 TI - Gastroenteritis: a nonclinical analysis. PMID- 11910955 TI - NORA (National Osteoporosis Risk Assessment) study sounds alarm on risk of osteoporotic fractures. PMID- 11910956 TI - Population-based effort takes promise of DM to a new level with CAD patients. AB - The case for early investments in DM for CAD patients. While the majority of DM programs tend to focus on the most severely ill patients, a strong case can be made that earlier intervention would offer greater benefits--at least over the long term. In fact, using such cutting-edge tools as predictive modeling, a handful of pioneering organizations are beginning to take their DM programs to this next stage, and early results show promise. PMID- 11910957 TI - Modest steps produce major results in initiative aimed at improving birth outcomes. AB - Consider strategies to boost early identification of pregnant members. Why? Because the earlier you can get women into prenatal care, the better your chances for positive birth outcomes. And, especially for many Medicaid plans, pregnancy related complications are high-ticket items. The good news is: identification strategies need not be particularly expensive or difficult to implement. See how one Chicago-based plan has been able to make great strides in this area in a short period of time. PMID- 11910958 TI - Recognize the subtleties of depression in primary care. AB - PCPs: learn how to recognize bipolar disorder in your depressed patients. Experts maintain that a high percentage of depressed patients who are treatment refractory may actually suffer from a milder form of bipolar disorder known as bipolar II. Not only is this condition much more prevalent than previously thought, it is often misdiagnosed, leading to poor outcomes and continuing utilization. While PCPs may not be comfortable prescribing the mood-stabilizing drugs generally indicated for bipolar cases, they can easily learn how to recognize the potential for this condition so patients can get the help they need quickly. PMID- 11910959 TI - Susceptibility of bacterial isolates to cefepime in comparison to other broad spectrum antimicrobial agents at a tertiary care center in Lebanon. AB - Newly introduced antimicrobial agents have to be evaluated to establish their current activity and susceptibility data base against microorganisms for future comparison. Cefepime is a fourth generation cephalosporin that was recently introduced in Lebanon but no background susceptibility data is available for it in this country. We prospectively analyzed the antimicrobial susceptibility pattern of the bacterial isolates from the American University of Beirut Medical Center to a number of broad spectrum antimicrobial agents and compared it to the susceptibility of cefepime. Consecutive clinical bacterial isolates, representing 665 gram-negative and 387 gram-positive were tested: 82 to 100% of the gram negative isolates were susceptible to cefepime, including most of the extended spectrum beta-lactamase producing Enterobacteriaceae. All the oxacillin susceptible staphylococcus isolates, and the penicillin susceptible Streptococcus spp. as well as 92% of the S. pneumoniae isolates were susceptible. The data shows that currently, cefepime provides a very broad spectrum of activity against gram-positive as well as gram-negative bacterial isolates recovered from clinical specimens. PMID- 11910960 TI - Approach to acute bacterial meningitis. Minireview. PMID- 11910961 TI - Nasoseptal variation in relation to sinusitis. A computerized tomographic evaluation. AB - Paraseptal structural abnormalities are common in patients with sinusitis. They may coexist with radiographic changes suggestive of sinusitis. Their etiological role in patients with no history of sinus disease is still controversial. In this study, computerized tomographic scan of orbits of 89 cases with no history of sinusitis were reviewed between 1996 and 1998. The incidence of septal deviation, concha bullosa and paradoxical middle turbinates was looked at. Their correlation with sinusitis was analyzed using CM-square statistical method. Results showed that the presence of these anatomical abnormalities is not associated with an increased incidence of sinusitis as shown radiologically. We conclude that the presence of septal deviation or large middle turbine on routine rhinoscopy does not mandate further radiological evaluation of the sinuses in the absence of history of sinusitis. PMID- 11910962 TI - Overview of the World Health Report 2000 Health systems: improving performance. PMID- 11910963 TI - Constitutional chromosome abnormalities among patients referred for blood karyotype analysis: a 5-year study at the AUBMC. AB - We report results on 2010 cases of blood referred for constitutional karyotype analysis. Referrals were grouped into 16 different categories, of which reproductive failure represented the highest percentage (33%), followed by structural congenital abnormalities (14.17%), developmental delay (11.34%), Down syndrome (9.65%), and abnormal sexual development (8.16%), while other categories represented smaller percentages. The total rate of abnormality was 16%, and the highest abnormality rates were among the clinically-recognizable chromosomal syndromes, while lower percentages were detected among less specific referrals. However, abnormality rates were generally different from the typical reported rates, probably due to the inclusion of cases not requiring chromosome analysis or the failure to recognize specific chromosomal syndromes. Other identified problems included lack of proper phenotypic description and difficulty in obtaining familial follow-up for proper diagnosis and genetic counseling. PMID- 11910964 TI - Portal hypertension. Update on pathophysiology and management. AB - A lot has been learned about the pathophysiology of portal hypertension and its complications. This knowledge has led to vast advancements in therapy for this serious disease. I believe the future will carry more breakthroughs in therapy intercepting earlier steps in the disease process such as arresting fibrogenesis, inducing fibrinolysis or possibly targeting vascular remodeling and neovessel formation or employing hepatocytes transplantation. PMID- 11910965 TI - Spontaneous bacterial peritonitis (SBP). PMID- 11910966 TI - Liver disease in pregnancy. PMID- 11910967 TI - Update on the status of liver transplantation in Lebanon. PMID- 11910968 TI - Hepatitis A: an updated overview. PMID- 11910969 TI - Hepatitis B virus infection. PMID- 11910970 TI - Hepatitis C. PMID- 11910971 TI - Hepatitis E, G, and TT virus. PMID- 11910972 TI - Autoimmune hepatitis. PMID- 11910973 TI - Non alcoholic steatohepatitis. PMID- 11910974 TI - Alcoholic liver disease. PMID- 11910975 TI - [Present and future of insulin therapy in type 2 diabetic patients]. AB - Type 2 diabetes natural history and progressive deterioration requires a therapeutic approach starting with diet and physical activity changes, then associated with pharmacological interventions (monotherapy, then multiple therapy). When oral anti-diabetics at optimal dosage have failed to maintain good diabetic control (HbA1c 6.5% Pounds), insulin treatment has to be considered and should be used when diabetic controls remain poor (HbA1c > 8%). The most adequate insulin regimen are bedtime NPH associated or not with oral anti-diabetics, or alternatively 2 daily injections (morning and evening) of intermediate insulin. Intensive insulin (3 daily injections or more), and particularly the use of short acting analogues is an effective regimen when bedtime or conventional regimen have failed. Insulin glargin could be an interesting alternative to bedtime NPH but needs further data in type 2 diabetes. Insulin treatment has to be initiated with an adequate patient (and family) education. Glycaemic objectives (fasting blood glucose and HbA1c) should be regularly assessed and adapted taking into consideration diabetic control, clinical effects and safety assessments (weight gain, hypoglycaemic events...). PMID- 11910976 TI - [Insulin and weight gain: myth or reality?]. AB - Most patients with type 2 diabetes gain weight when treated with insulin. Weight gain is observed when insulin is introduced after oral agents have failed, but also when insulin is introduced shortly after the diagnosis of diabetes. The mechanisms of this weight gain are incompletely understood, but reduction of energy lost by glucosuria and reduction of energy needed for glucose production are main determinants. The same reasons apply to the weight gain observed at the beginning of treatment with sulfonylureas, even though patients usually gain less weight with sulfonyulreas than with insulin. In the UKPDS, at 10 years of the study, patients treated with insulin gained 2 kg more, i.e. 2.5% of the average weight of patients included in the trial, than patients treated with sulfonylureas. The reasons of the excess of weight gain with insulin as compared with sulfonylureas remain unclear. Patients with type 2 diabetes treated with insulin gain weight only during the first 2-3 years after insulin introduction. The weight stabilizes thereafter. Type 2 diabetes usually remains unknown for years before diagnosis and patients may lose weight during this long period of time preceding diagnosis. It is hypothesized that the weight gain observed after the introduction of insulin may simply correspond to the reexpression of the physiologically controlled body weight. PMID- 11910977 TI - [Inhaled insulin: clinical results in type 2 diabetic patients]. AB - French type 2 diabetic patients are underinsulinised mainly because of fear of injections. Among all alternatives to the subcutaneous route, only the lung has a sufficient bioavailability of 10% without addition of promoters. This is made possible by the large surface and thinness of the alveolo-capillary barrier, and only if insulin particles have an ideal size (2-3 microns). The Pfizer-Aventis Inhale project utilises powder aerosolisation, whether the Novo-Aradigm project utilises liquid nebulization. In the former project, phase 1 studies have shown plasma kinetics similar to subcutaneous lispro, and intrasubject reproducibility non significantly different from rapid and lispro insulins. Phase 2 studies, performed on more than 200 insulin-treated and non insulin-treated subjects have shown an efficacy similar to subcutaneous insulin, with no difference in terms of side-effects (hypoglycaemia, weight gain) and a satisfactory 1-year local tolerance, as evaluated by functional tests. Phase 3 studies, performed on 1400 subjects have just been completed and are not published yet. Though results are promising, some important questions remain to be clarified: long term tolerance, miniaturisation of the inhaler, overcost, and long-term acceptability, especially in type 2 patients. It already appears that the major potential indications may be "functional" (flexible) insulin therapy of type 1 diabetes and early insulinisation of type 2 patients with oral drugs failure. PMID- 11910978 TI - [Towards earlier nd more aggressive insulin therapy in type 2 diabetic patients]. PMID- 11910979 TI - [Insulin treatment in type 2 diabetes: epidemiological data]. AB - After several years of disease duration, blood glucose control is difficult without using insulin in type 2 diabetic patients. However, large differences are observed regarding the use of insulin from the south to the north of Europe. A positive effect of insulin therapy on metabolic control compared to oral antidiabetic agents is shown only in clinical trials performed on selected patients. As to degenerative complications, a great excess of retinopathy is observed in insulin-treated patients in several surveys, without possibility to determine the specific effect of treatment from that of disease severity. The UKPDS is the only randomised long-term clinical trial with a large number of patients. The aim was to compare conventional to intensive antidiabetic treatment, whatever the type or drug, insulin or sulfonyureas. After a ten-year follow-up, glycemic control was better with the intensive treatment, the microvascular complications less frequent by 25%, but the benefit on cardiovascular complications or mortality was modest. Another clinical trial from US shows that a too strict insulin regimen could perhaps have no effect or even opposite to the objectives regarding cardiovascular morbidity. Considering these clinical uncertainties and the need for more definite long-term clinical data, it is justified to conduct additional long-term clinical studies in this field. Furthermore after secondary failure of oral anti-diabetic agents in type 2 diabetic patients, it appears reasonable to discuss the rational for insulin treatment based on an individual assessment, particularly in older diabetics, weighing the whole benefits and risks for a given patient of such an important therapeutic change. PMID- 11910980 TI - [Modifications in myocardial energy metabolism in diabetic patients]]. AB - The capacity of cardiac myocyte to regulate ATP production to face any change in energy demand is a major determinant of cardiac function. Because FA is the main heart fuel (although the most expensive one in oxygen, and prompt to induce deleterious effects), this process is based on a balanced fatty acid (FA) metabolism. Several pathological situations are associated with an accumulation of FA or derivatives, or with an excessive b-oxidation. The diabetic cardiomyocyte is characterised by an over consumption of FA. The control of the FA/glucose balance clearly appears as a new strategy for cytoprotection, particularly in diabetes and requires a reduced FA contribution to ATP production. Cardiac myocytes can control FA mitochondrial entry, but display weak ability to control FA uptake, thus the fate of non beta-oxidized FA appear as a new impairment for the cell. Both the trigger and the regulation of cardiac contraction result from membrane activity, and the other major FA function in the myocardium is their role in membrane homeostasis, through the phospholipid synthesis and remodeling pathways. Sudden death, hypercatecholaminemia, diabetes and heart failure have been associated with an altered PUFA content in cardiac membranes. Experimental data suggest that the 2 metabolic pathways involved in membrane homeostasis may represent therapeutic targets for cytoprotection. The drugs that increase cardiac phospholipid turnover (trimetazidine, ranolazine,...) display anti-ischemic non hemodynamic effect. This effect is based on a redirection of FA utilization towards phospholipid synthesis, which decrease their availability for energy production. A nutritional approach gave also promising results. Besides its anti-arrhythmic effect, the dietary docosahexaenoic acid is able to reduce FA energy consumption and hence oxygen demand. The cardiac metabolic pathways involving FA should be considered as a whole, precariously balanced. The diabetic heart being characterised by a different metabolic "status" with similarities to that of myocardium in coronary disease. Diabetes and other chronic cardiac diseases share common FA metabolism disorders leading to an altered energy balance, a decrease in long chain polyunsaturated Fas, and altered FA profiles in cardiac membranes. These disturbances, however, do not represent independent therapeutic targets, and should be considered as a whole. PMID- 11910981 TI - [Clinical and diagnostic aspects of coronary disease in diabetic patients]. AB - Coronary heart disease in diabetic patients is more frequent than in non-diabetic patients, even without other associated risk factors. It is generally characterised by multi-vessel and distal lesions, associated with an increase incidence of moderate diffuse stenoses, endothelial dysfunction and intramyocardial microangiopathy. Diabetic patients present with a higher incidence of silent myocardial ischaemia (around 30%) than non-diabetic patients. The diagnosis of coronary artery disease has to be assessed mainly using non invasive techniques, particularly cardiac imaging with sensitising tests: thallium myocardial scintigraphy, echocardiography using exercise stress test or pharmacologically-induced stress (dipyridanole, dobutamide). The characteristics of these methods and their predictive values are described, and a step-by-step diagnosis approach is proposed. PMID- 11910983 TI - [The heart is at the heart of the problem of the management of type 2 diabetic patients]. PMID- 11910982 TI - [Metabolic considerations in the treatment of coronary disease in diabetic patients]. AB - Coronary heart disease represents the first cause of death in diabetic patients. The poor outcome of this ischemic disease is multifactorial. The abnormalities in myocardial energy metabolism encountered by the diabetic heart explain both the ischemic severity and the worsening of reperfusion lesions. The metabolic abnormalities in diabetic heart consist in both an impairment of glucose metabolism (diminished glucose uptake, reduced glycolysis, decreased glucose oxidation...) and an increase in fatty acid oxidation. During ischemia, glucose oxidation is reduced and anaerobic glycolysis becomes the main ATP substrate. Lactate accumulate in myocardial cells, inducing both a metabolic acidosis and an intracellular calcium overload. During reperfusion, intracellular homeostasis is restored very slowly in diabetic heart. Several therapeutic approaches are used to correct these metabolic disturbances. Glucose--insulin--potassium infusion in acute myocardial infarction leads to a significant reduction in the mortality relative risk in diabetic patients (ECLA and DIGAMI studies). The benefit is greater in diabetic patients who where non-insulin-treated prior to ischemia followed by myocardial reperfusion therapy. Others more direct pharmacological approaches improve glucose oxidation during myocardial ischaemia and myocardial reperfusion. The reference drug remains trimetazidine for which one of the fundamental mechanism of action was discovered recently. This specific metabolic, non haemodynamic approach, complete the gold-standard treatment of coronary heart disease in diabetic patients (e.g. aspirin, beta-blockers, ACE inhibitors and statins). PMID- 11910984 TI - [Epidemiology of heart disease in diabetes]. AB - Diabetes mellitus is a frequent disease--3% of the French population in 1999. Vital prognosis is essentially related to cardiac complications, at least in type 2 diabetic patients, that is 85 to 90% of diabetic subjects. The most frequently observed cardiac complications are symptomatic or silent ischemic coronary disease and myocardial events, followed by heart failure. These complications may also result from cardiac autonomic neuropathy, be manifested by arrhythmia and even lead to sudden death. Data on the prevalence and incidence of these events are scarce, and the epidemiologic characteristics are not the primary aim of publications. This review analyses: cardiovascular drug consumption in French diabetic and non diabetic patients, the incidence and prevalence rates of the most important cardiac complications from some large general prospective studies which included enough diabetic subjects, coronographic severity scores for diabetic and non-diabetic subjects, prognosis following treated coronary events, some prevalence data on cardiac autonomic neuropathy. PMID- 11910985 TI - [Fatigue in patients with chronic hepatitis C: a sign to look for]. PMID- 11910986 TI - [Hepavir, the first observational study of one cohort of patients treated with alpha-2a interferon, monotherapy. Evaluation of asthenia and its social consequences]. AB - OBJECTIVES: The aim of this observational study in patients with chronic hepatitis C and treated with interferon alpha-2a was to assess 1) monitoring in everyday practice, 2) the acceptability of treatment and 3) the intensity of fatigue. METHODS: Three hundred and fifty four patients were enrolled by physicians in both teaching and general hospitals, or private practice. Before treatment, clinical, epidemiological, and virological data were collected as well as a self-evaluation of fatigue using a visual analogic scale. Clinical follow-up was assessed every 3 months during treatment and 6 months after the end of treatment and included an evaluation of fatigue and the number of workdays missed due to sickness. RESULTS: Two hundred and nineteen men and 135 women, mean age 45 +/- 13, were included. The epidemiological, histological and virological features of this group were similar to those patients usually treated for chronic hepatitis C. Before treatment, the mean measurement of fatigue was 41 on a scale from 0 (perfect form) to 100 (exhausted). Fatigue was unrelated to age, source of infection, biological activity, or histological score. It worsened in patients who stopped interferon after 3 or 6 months, but was stable in patients who continued treatment for 12 months. Fatigue decreased after the end of treatment and was unrelated to treatment response. The need to stop work was strongly related to the intensity of fatigue and the number of workdays missed due to sickness represented nearly two months out of three in 25% of active patients during the first quarter and in 15% of patients thereafter. 61% of patients self injected interferon (mainly previous drug users) whereas 30% of patients used nurse care throughout treatment. CONCLUSION: This study not only provides a realistic evaluation of fatigue in patients with chronic hepatitis C, before, during and after treatment, but also highlights its social and economic consequences. It shows the need for further cost-effectiveness studies on new therapeutic strategies using combined treatments. PMID- 11910987 TI - [Genetic alterations in hepatocellular carcinomas: associations with clinical parameters]. PMID- 11910988 TI - [Factors of virulence of Helicobacter pylori, what is it the importance?]. PMID- 11910989 TI - [The irresistible adventure of laparoscopic colorectal surgery]. PMID- 11910991 TI - Vasoactive intestinal peptide is involved in the inhibitory effect of interleukin 1 beta on the jejunal contractile response induced by acetylcholine. AB - Although previous studies have shown that interleukin-1 beta (IL-1 beta) decreases acetylcholine (ACh)-induced intestinal contraction by an action on the enteric nervous system, the neuromediator(s) involved are still unknown. AIM: To determine the role of nitric oxide (NO), vasoactive intestinal peptide (VIP) and/or adenosine triphosphate (ATP) in mediating this inhibitory effect. METHODS: The effects of NO synthase inhibitors, VIP and ATP antagonists on motor response to the ACh were investigated before and after 90-min exposure of a rat preparation of jejunal longitudinal muscle-myenteric plexus to IL-1 beta. NG nitro-L-arginine methyl ester, NG-nitro-L-arginine and NG-monomethyl-L-arginine were used to inhibit NO synthase, VIP (10-28) and [D-p-Cl-Phe6, Leu17] VIP to block VIP receptors, and suramin to block ATP receptors. RESULTS: NO synthase inhibitors failed to block the inhibitory effect of IL-1 beta on ACh-contracted jejunum smooth muscle. Suramin also failed to affect IL-1 beta-induced inhibition, whereas VIP antagonists abolished it. Moreover, the action of IL-1 beta was partly reproduced by VIP. CONCLUSIONS: While neither NO nor ATP accounts for the inhibitory effect of IL-1 beta on ACh-contracted jejunum, VIP seems to be a key-mediator of this effect. PMID- 11910990 TI - [CagA status and virulence of Helicobacter pylori strains. Results of a French multicentric prospective study]. AB - Previous experimental and epidemiological studies with few patients suggested that the presence of the cagA gene was a virulence factor for Helicobacter pylori (H. pylori). AIM: To establish in this large epidemiological cohort study the relationship between the histological virulence of H. pylori infection and the cagA status of the bacteria. METHODS: This prospective cohort study (6 month follow-up) was conducted on adult patients undergoing endoscopy for upper gastrointestinal symptoms. The cagA status of H. pylori-positive patients was established using the polymerase chain reaction (PCR) method on an antral biopsy. A score of histological virulence (inflammation, activity) was recorded on the basis of the Sydney system (on antral, angular and fundic biopsies). Eradication treatment given was not imposed and a clinical follow-up was performed at 3 and 6 months. H. pylori eradication was verified by a 13C urea breath test at 3 months. RESULTS: Four hundred and twenty two centers recruited 652 patients (mean age: 51 +/- 15 years, 55% female). Upper GI endoscopy was abnormal in 80% of the patients of whom 68% had a gastritis aspect; 38% were infected by H. pylori, and among them 51% were cagA-positive. The histological virulence scores associated with the cagA-positive strains were significantly higher than those associated with the cagA-negative strains, globally (P = 0.0035), in the antrum (P = 0.0063), and in the angulus (P = 0.046), but not in the fundus (P = 0.05). The cagA status was correlated neither with the symptom severity at inclusion and at 6 months (P > 0.05), nor with the H. pylori eradication rate at 3 months (75% in cagA-positive and 70% in cagA-negative strains, P = 0.52). CONCLUSION: This study on a large cohort of patients confirms the greater histological virulence of H. pylori cagA positive strains. However, this virulence was not associated with more severe symptoms nor with an increase in resistance to H. pylori eradication treatment. PMID- 11910992 TI - [Laparoscopic colorectal surgery in the era of evidence- based medicine]. PMID- 11910993 TI - [Epithelioid hemangioendothelioma associated with nodular regenerative hyperplasia]. AB - Epithelioid hemangioendothelioma of the liver is a rare neoplasm of vascular origin. We report a case of epithelioid hemangioendothelioma occurring in a patient with nodular regenerative hyperplasia. This association suggests that some hepatic vascular changes might promote the growth of epithelioid hemangioendothelioma. PMID- 11910995 TI - [Liver transplantation in a patient with cirrhosis and an uncontrolled extra hepatic infectious focus]. PMID- 11910994 TI - [Lanreotide acetate may cure cystic dystrophy in heterotopic pancreas of the duodenal wall]. AB - Cystic dystrophy in heterotopic pancreas of the duodenal wall is a rare but benign disease, associated in most of the cases with chronic pancreatitis. Treatment of this disease is controversial. We report here the use of a long acting somatostatin synthetic stable analogue in the treatment of a cystic dystrophy in heterotopic pancreas of the duodenal wall: a 45-year-old man, hard drinker, was treated successfully during three months with lanreotide acetate; disappearance of cysts was confirmed by a computed tomography two months after the end of treatment. PMID- 11910997 TI - [Hepatotoxicity of mizolastine (Mizollen) : report of 2 cases]. PMID- 11910996 TI - [Liver abscesses due to Yersinia pseudotuberculosis discloses a genetic hemochromatosis]. PMID- 11910998 TI - [Primary malignant melanoma of the esophagus: value of endoscopic ultrasonography in the assessment of extension]. PMID- 11910999 TI - [Acute pancreatitis by pancreatico-gastric anastomosis stenosis three years after pancreaticoduodenectomy]. PMID- 11911000 TI - [Surgical management of pancreatic metastasis from renal cell carcinoma]. PMID- 11911001 TI - [Pneumomediastinum without colonic perforation during a severe attack of ulcerative colitis]. PMID- 11911002 TI - [Colonic cancer and pneumococcal bacteremia:an unusual association]. PMID- 11911003 TI - [Hepatic injuries related to Lyme borreliosis: response to 2 cases presented by I. Dadamessi et al]. PMID- 11911004 TI - [Helicobacter pylori increases the antisecretory activity of omeprazole in the parietal cell and the proton pump]. PMID- 11911005 TI - [Is preoperative radiotherapy necessary for rectal cancer treated by total resection of the mesorectum?]. PMID- 11911006 TI - Measuring teacher attitudes and expectations toward students with ADHD: development of the test of knowledge about ADHD (KADD). AB - OBJECTIVE: The attitudes, expectations, and behaviors of teachers toward children with Attention-Deficit/Hyperactivity Disorder (ADHD) may have a lasting impact on the academic self-efficacy and success of students with ADHD. Yet, to date there exists no standardized measure of teacher attitudes or expectations toward children with ADHD. The purpose of this study was to develop an instrument to measure teacher attitudes and expectancies toward children with ADHD. METHOD: The Test of Knowledge About ADHD (KADD) was constructed based on the error-choice method, which is an indirect method of attitude measurement. This psychometric approach was utilized to reduce patterns of response distortions that produce systematic errors in direct attitudinal assessment. Elementary school teachers (N = 103) completed the error-choice instrument. CONCLUSION: Results indicated adequate to good internal consistency reliability for the KADD for this sample, with a Cronbach's coefficient alpha of 0.82. Additionally, results yielded initial discriminant evidence to suggest that the KADD has construct validity for this sample. Implications for the use of this instrument in applied areas are discussed. PMID- 11911008 TI - Validity of the Brown ADD scales: an investigation in a predominantly inattentive ADHD adolescent sample with and without reading disabilities. AB - The Brown ADD Scale for Adolescents is widely used in clinical settings, yet, no published studies have investigated divergent and concurrent validity and specificity and sensitivity to inattentive ADHD symptomatology. Ninety-eight participants (13 to 16 years) were classified as ADHD/I and/or reading disabled (RD) using Kiddie Schedule for Affective Disorder and Schizophrenia (K-SADS), Conners' Rating Scales (CRS-R), and Ontario Child Health Study Scales (OCHSS), WRAT3, and WRMT-R. The results were: 29 ADHD/I; 12 RD, 16 ADHD/I with RD; and, 41 controls. The RD group was included to evaluate specificity. The Brown was administered but not used in classification. The ADHD groups scored higher on the Brown subscales compared with the other two groups. The recommended cutoffs resulted in high rates of false negatives but few false positives; this suggests good specificity but poor sensitivity. There were moderate correlations among the Brown, CRS-R, and OCHSS. The Brown can be useful in screening out ADHD; however, its low sensitivity precludes its usefulness in diagnosing ADHD. PMID- 11911007 TI - A meta-analytic review of gender differences in ADHD. AB - The present study examined gender differences in ADHD through a meta-analysis. Effect size estimates for the primary symptoms and correlates of ADHD were calculated in an attempt to replicate and extend a previous meta-analysis on gender differences in the disorder. Relatively lenient inclusion criteria were used in order to maximize the number of studies included in the effect sizes. The results indicated that in comparison to ADHD boys, ADHD girls had lower ratings on hyperactivity, inattention, impulsivity, and externalizing problems. In addition, ADHD girls had greater intellectual impairments and more internalizing problems than ADHD boys. Overall, the results of the current meta-analysis indicated general agreement with the previous meta-analysis. The clinical implications of these gender differences and future research considerations are discussed. PMID- 11911009 TI - Psychosocial adjustment and peer competence of siblings of children with Attention-Deficit/Hyperactivity Disorder. AB - Preliminary data examined the relationships between mothers, their sons with Attention-Deficit/Hyperactivity Disorder (ADHD), and younger siblings. We hypothesized that the conflict between the mother and the son with ADHD would also occur in the relationship between the child with ADHD and the younger sibling. Significant associations were found among ADHD behaviors, family conflict variables, conflict in the relationship between mother and child with ADHD, conflict in the relationship between mother and younger sibling, and conflict in the relationship between the child with ADHD and the younger sibling. Further, significant associations were found between behavior symptoms associated with ADHD and more problematic family relationships. Of particular interest was the finding of an unexpected inverse association between siblings' peer competence and mother-ADHD conflict; specifically, that conflict occurring at home from ADHD accounted for a greater percentage of the variance in peer competence as rated by teachers. Recommendations are made for future research with larger samples, alternative designs, older siblings, and fathers. PMID- 11911011 TI - Manpower and medicine. PMID- 11911010 TI - Audits, investigations and serious trouble. PMID- 11911012 TI - Heart transplants: need versus availability. AB - Every year in the US heart failure accounts for roughly 60,000 deaths and is the contributing cause in another 300,000 deaths. The two-year survival rate for patients with advanced heart failure is less than 50%, with the incidence of death at 106 in 100,000, more than that for AIDS and breast cancer combined. As these figures attest, the economic burden is quite extensive. The Centers for Medicaid and Medicare estimate a cost of $10 billion a year for this diagnosis alone. Both the human and financial cost have impelled doctors and researchers to improve their capacity to treat heart failure both through conventional methods and, in the most serious cases, through transplantation. Many pioneers have either directly or indirectly contributed to our ability to treat heart failure. Among these early researchers were: Dr Alexis Carrel, who was awarded the Nobel Prize for his pioneering work in vascular anastomosis; Dr John Gibbon, who did important work in the development of the cardiopulmonary bypass machine; Drs Normal Shumway, Richard Lower, and Demikhov, who developed heart transplant procedures in the canine model; Dr Christian Barnaard, who performed the first technically successful human-to-human heart transplant (1967); and Dr Thomas Hardy, who attempted the first xenotransplant (1963). While these achievements were phenomenal advances, long-term survival for transplant recipients was minimal until progress was made in immunosuppressive techniques. PMID- 11911013 TI - Geographic variation in breast-conserving surgery in Kentucky's Medicare population. AB - Breast cancer represents a significant disease burden in Kentucky, affecting some 2,900 newly diagnosed women each year. About 600 Kentucky women will die of breast cancer in 2001. Kentucky's age-adjusted death rate for breast cancer in 1995 was 24.2 per 100,000 women, ranking 30th in the United States. In Kentucky and nationally, it is known that breast cancer treatment and prognosis may be complicated by elderly women's age and comorbid illnesses. Not all differences in treatment and prognosis can be explained, however, by patient characteristics or illness severity. A large body of health services research has developed over the last thirty years that documents the relationship between the supply of health care resources (e.g., physicians, hospital beds) and variances in treatment. This line of research has particular significance in Kentucky, where the supply of resources varies greatly across the state. Because breast cancer treatment variations unrelated to breast cancer disease may affect prognosis and outcome, it is important that these treatment variations be understood. The purpose of this study was to determine the existence and extent to which Kentucky women over 65 with the same stage of breast cancer receive breast-conserving surgery in different geographic regions, and to link geographic differences to differences in population characteristics, and the availability of health care resources for cancer treatment. PMID- 11911014 TI - gamma-Hydroxybutyric acid and 5-fluorouracil, metabolites of UFT, inhibit the angiogenesis induced by vascular endothelial growth factor. AB - UFT, a drug composed of uracil and tegafur at the molar ratio of 4:1, is an orally active agent for the treatment of a wide variety of malignant tumours. Using a murine dorsal air sac (DAS) assay, we have previously shown that UFT and its metabolites, gamma-hydroxybutyric acid (GHB) and 5-fluorouracil (5-FU), inhibited the angiogenesis induced by murine renal cell carcinoma. Here we report that UFT was more effective than other fluorinated pyrimidines such as 5-FU and doxifluridine (5'-DFUR) in blocking the angiogenic responses elicited by five human cancer cell lines which produced high levels of vascular endothelial growth factor (VEGF), but no detectable fibroblast growth factor-2 (FGF-2) in vitro. In contrast, UFT was unable to block the angiogenic response to one human gastric cancer cell line which produced both VEGF and FGF-2 in vitro. However, the production or secretion of VEGF by these cells was unaffected by GHB and 5-FU treatment. Interestingly, GHB suppressed the chemotactic migration and tube formation of human umbilical vein endothelial cells (HUVECs) stimulated by VEGF, without inhibiting their DNA synthesis. Since GHB did not affect the FGF-2-driven activities in HUVECs, its action appears to be VEGF-selective. On the other hand, 5-FU inhibited DNA synthesis and migration of HUVECs stimulated by both VEGF and FGF-2, and tube formation driven by VEGF, suggesting that 5-FU is cytotoxic to endothelial cells. The inhibitory effects of 5-FU, and especially those GHB, were reproduced under in vivo condition using the DAS assay. The VEGF-mediated angiogenesis was significantly inhibited by UFT, 5-FU, and especially by GHB. We propose that the selective inhibitory effects of GHB on VEGF-mediated responses of endothelial cells are involved in the anti-angiogenic activity of UFT. PMID- 11911015 TI - Inhibition of human angiogenesis with heparin and hydrocortisone. AB - Angiogenesis is a critical determinant of tumor growth and the development of metastases. Heparin, steroids, and heparin/steroid combinations have been used in a variety of in vitro models and in vivo in animal models as effective inhibitors of angiogenesis. We tested heparin, steroid and heparin/steroid combinations at a variety of concentrations to determine their effect on the human 'angiogenic switch' from a resting to a proliferative endothelium in vessels from three placentas (initiation), and the effect of these compounds on the subsequent growth of a human angiogenic response (promotion). Using full-thickness human placental vein discs cultured in three-dimensional fibrin-thrombin clots, we demonstrated that heparin (300, 3000 micrograms/ml), steroid (350, 3500 micrograms/ml), and combinations of heparin/steroid at these doses effectively blocked both initiation and promotion of a human angiogenic response in a dose dependent fashion. We also demonstrated that high-dose steroid or heparin/steroid treatment for 15 days resulted in disruption of vessel integrity, while treatment with heparin alone produced a suppressed growth rate but had intact vessel architecture. High-dose heparin/steroid treatment could also disrupt a developed angiogenic response and retard further development of an angiogenic response following the cessation of treatment. PMID- 11911016 TI - Quantitative analysis of angiogenesis using confocal laser scanning microscopy. AB - Angiogenesis is essential for tumor growth and metastasis. Angiogenesis is commonly quantified by measuring microvessel density (MVD) within tumors. In this report, we compared light microscopy with confocal laser scanning microscopy (CLSM) in the qualitative and quantitative analysis of angiogenesis. MVDs were determined manually in a lung tumor xenograft and a normal skeletal muscle using CD31 immunohistochemical staining and light microscopy. Area of three-dimensional representation of microvessels, detected as CD31 immunofluorescence, was measured automatically using computer-assisted CLSM. By manual counting under light microscopy, the relative level of MVD of the lung tumor vs. skeletal muscle was 0.8. However, the corresponding relative level of microvessels was 3.4 as determined by computer-assisted CLSM. Furthermore, the architecture of microvessels was better delineated with CLSM than with light microscopy. We have applied this CLSM method for analyzing the antiangiogenic effect of an anticancer drug, paclitaxel, in the lung tumor xenograft model. We conclude that CLSM is an appropriate method for quantitative and qualitative analysis of microvasculature in normal and tumor tissues. PMID- 11911017 TI - Endostatin inhibits angiogenesis by stabilization of newly formed endothelial tubes. AB - Endostatin decreased vascular endothelial growth factor (VEGF)-induced formation of endothelial tubes and microvessels sprouting from aortic rings and blocked their network. After cessation of treatment, the survival time of endostatin plus VEGF-treated tubes was approximately doubled in comparison to VEGF alone. Endostatin antibody blocked VEGF-induced endothelial tube formation and disrupted existing tubes. Endostatin immunostaining was localized between endothelium and basement membrane and in inter-endothelial junctions of new, but not of quiescent, blood vessels. In tumors grown in SCID mice, endostatin immunostaining was stronger accompanying blood vessel maturation and was significantly prominent in vessels of tumor marginal zone where angiogenesis is highly active. These data indicate a new antiangiogenic action of endostatin stabilizing and maturating endothelial tubes of newly formed blood vessels. Thus, strategies accelerating vascular stabilization and maturation could be promising in tumor therapy. PMID- 11911018 TI - Case report: bronchiolitis obliterans organizing pneumonia in a spice process technician. PMID- 11911019 TI - Rapidly progressive, fixed airway obstructive disease in popcorn workers: a new occupational pulmonary illness? PMID- 11911020 TI - A follow-up: high level of dioxin contamination in Vietnamese from agent orange, three decades after the end of spraying. PMID- 11911021 TI - Hepatitis C virus infection among public safety workers. PMID- 11911022 TI - Hepatitis C screening and prevalence among urban and rural public safety workers in Oregon. PMID- 11911023 TI - The incidence of green tobacco sickness among Latino farmworkers. PMID- 11911024 TI - Effect of the Heart At Work program on awareness of risk factors, self-efficacy, and health behaviors. AB - This study evaluates the effects of the American Heart Association's Heart At Work program on cardiovascular disease risk factor awareness, self-efficacy, and health behaviors. A prospective, quasi-experimental research design was used to assess the impact of the program at two factory sites (one intervention and one control). A total of 633 employees participated. Intervention site respondents significantly improved their knowledge of blood pressure management, the relationship between nutrition and cardiovascular disease, and heart attack risk factors. They also were more likely to begin treatment for hypertension, to report fewer sick days, and to have plans to improve their diet and lose weight. These findings suggest that the Heart At Work program had a favorable overall impact. PMID- 11911026 TI - Evaluation of the North Country on the Job Network: a model of facilitated care for injured workers in rural upstate New York. AB - We describe the evaluation of a community-based program designed to facilitate access to care and return to work for injured workers in a rural, medically underserved area in upstate New York. Providers are recruited to provide easily accessible care and are oriented to concepts of transitional duty and rapid return to work as medically appropriate; companies are recruited with the agreement to provide transitional work for injured employees. Registered nurses, hired by the local hospital, serve as case coordinators to facilitate care and coordinate communications among all parties. Over 3000 injured workers received care through the program in the first 56 months, with a decline in the number of transitional days over time. The number of days that the cases remain open has steadily declined, and the number of return-to-work cases has increased. The success of this initiative provides an excellent background for continued improvement in delivery of care to injured workers and proactive efforts at improving workplace safety and health in a rural area. PMID- 11911025 TI - Impact of case manager training on worksite accommodations in workers' compensation claimants with upper extremity disorders. AB - Management of the return-to-work process in claimants with work-related upper extremity disorders often poses challenges to the health care provider, claimant, and employer. Modifying workplace ergonomic risk factors as a component of the workplace accommodation process may improve return-to-work outcomes by reducing recurrent pain and discomfort. The present study is a case-control evaluation of the effects of a 2-day training program for nurse case managers that was designed to facilitate the implementation of workplace accommodations within a workers' compensation health care delivery system. After the training, 101 claimants with compensable upper extremity disorders were randomly assigned to case managers with and without training. Overall, 208 accommodations were recommended and 155 of these were implemented (75%). Claimants of trained nurses received 1.5 times as many recommendations for accommodations as claimants managed by nurses not trained in the process, and 1.4 times as many accommodations were implemented, although no differences were found between the two groups in implementation rates. Trained nurses were more likely to recommend accommodations addressing workstation layout, computer-related improvements, furnishings, accessories, and lifting/carrying aids, whereas the untrained nurses were more likely to suggest light duty and lifting restrictions. This study indicates that the training was associated with a change in the practice behavior of case managers regarding the workplace accommodation process. More research is needed to identify barriers to implementation and develop more effective approaches to facilitate worksite accommodations in disabled workers with carpal tunnel syndrome and other persistent upper extremity disorders. PMID- 11911027 TI - Exposures in the painting trades and paint manufacturing industry and risk of cancer among men and women in Sweden. AB - Using data from the 1960 and 1970 Swedish censuses and the Swedish Cancer Register for 1971 to 1989, this study investigated variations in cancer risk by gender associated with employment in painting trades and paint manufacturing. Among men, standardized incidence ratios were significantly increased for lung cancer among painters and lacquerers; bladder cancer among artists; and pancreas cancer, lung cancer, and nonlymphocytic leukemia among paint and varnish plant workers. Risks for women were elevated for cancers of the esophagus, larynx, and oral cavity among lacquerers and for oral cancer among glaziers. These findings are consistent with the report of the International Agency for Research on Cancer that classified painting as an occupationally related cause of cancer and provide further evidence that the risk of certain cancers is increased by exposures in the paint manufacturing process. PMID- 11911028 TI - One-year study of occupational human immunodeficiency virus postexposure prophylaxis. AB - A 12-month experience with human immunodeficiency virus (HIV) postexposure prophylaxis (PEP) in a tertiary care center was evaluated for timeliness of treatment and adherence to treatment recommendations. Forty-six health care workers were started on HIV PEP. Risk status of the source patient, rather than type of exposure, was a significant determinant for both initiating and completing treatment. Of those exposed to HIV-positive sources, 79% completed the full 28 days of therapy. Only 22% of all health care workers who started PEP discontinued treatment because of adverse effects. Excluding three cases with significant delays in reporting and one in which treatment was controversial, the mean time from exposure to first dose of PEP was 1 hour and 46 minutes. The use of a defined treatment protocol, with supporting educational material and PEP medication immediately available, is an effective way of managing HIV exposures. PMID- 11911029 TI - Risk factors for multisymptom illness in US Army veterans of the Gulf War. AB - This research study examined the prevalence of symptoms and identified risk factors for reported symptoms among a group of Army Gulf War (GW) veterans. A survey was mailed to all members of the Ft. Devens cohort in 1997, representing the third assessment of a group that consisted of 2949 US Army soldiers deployed to the Gulf, and was studied initially in 1991. A total of 1290 subjects responded to the mailed survey; aggressive follow-up methods to address non response bias were employed. Subjects were classified as having multisymptom illness if they reported symptoms from at least two of three symptom categories (fatigue, mood-cognition, musculoskeletal). Sixty percent of the respondents met criteria for multisymptom illness. Female gender, lower levels of education, psychological symptoms, self-reported use of a medical clinic in the Gulf, ingestion of anti-nerve gas pills (pyridostigmine bromide), anthrax vaccination, tent heaters, exposure to oil fire smoke, and chemical odors were significantly related to multisymptom illness in logistic regression analyses. Analyses in which subjects were stratified by level of psychological symptoms revealed different sets of GW-service environmental exposures and suggest that subgroups of GW veterans may have different sets of risk factors. PMID- 11911030 TI - A longitudinal follow-up of pulmonary function and respiratory symptoms in workers exposed to manganese. AB - The purpose of the investigation was to study the effects on the respiratory system in mine workers with long-term exposure to manganese (Mn) in the workplace. The study included a follow-up of pulmonary function and respiratory symptoms among 145 workers employed in a large Mn mine and 65 matched controls, and the assessment of Mn concentrations in environment and biological samples. Lung function was measured by recording spirometric parameters. The Mn-exposed workers reported more respiratory symptoms and a significantly higher prevalence of all grades of pulmonary function impairment. All predicted symptoms except for asthma increased significantly in the current smoking group compared with the non smoking group. There was a significant decrease in FEV1, FVC, and FEV1% values in exposed workers compared with controls at stages 2 and 3, with an additive effect of the smoking habit. The Mn concentrations in blood, urine, and hair were significantly higher in the exposed workers than in the controls. The level of cumulative exposure index of workplace Mn was notable and did not change significantly over this study. The respiratory effects found in Mn-exposed workers were probably caused by the Mn in the workplace and the synergistic effect of smoking. These effects indicate a need for respiratory protection and improvements in the work environment. PMID- 11911031 TI - Elevated mortality from lung cancer associated with arsenic exposure for a limited duration. AB - In 1959, arsenic poisoning was detected in the town of Nakajo in Japan. The cause was exposure to inorganic arsenic in well water during 1954 to 1959. To examine the long-term effects of limited-duration arsenic exposure, we conducted mortality and survival studies for patients with chronic arsenic exposure and for control subjects from 1959 to 1992. The ratio of observed deaths to expected deaths from lung cancer was significantly high (7:0.64) for male patients. The lung cancer mortality rate was elevated markedly in subgroups with higher clinical severities of symptoms. Small cell carcinoma was specific to the exposed patients. The cumulative change of survival declined significantly in the exposed patients compared with the controls. The decline disappeared when lung cancer deaths were treated as lost to follow-up. The results showed that a 5-year period of arsenic exposure was associated with risk of lung cancer. PMID- 11911032 TI - Correlation between serum lipid profiles and the ratio and count of the CD16+ monocyte subset in peripheral blood of apparently healthy adults. AB - BACKGROUND AND PURPOSE: Deposited vascular oxidized low-density lipoproteins (LDLs) are important triggers of the transformation of circulatory monocytes into macrophages. CD16+ monocytes have been reported to be the precursors of tissue macrophages. In this study, we sought to determine the relationship between serum LDL-cholesterol and the percentage and count of the CD16+ monocyte subset in the peripheral blood of healthy adults. METHODS: We studied the correlations between serum lipid profiles and both peripheral CD16+ and CD36+ monocyte subset ratios and counts in apparently healthy adults (50 men and 50 women). Monocyte surface antigens CD16 and CD36 on CD14+ monocytes were detected using fluorescent triple staining and flow-cytometry. Surface staining was performed by incubating 1 x 10(6) blood mononuclear cells with phycocrythrin-conjugated anti-CD14, fluorescein isothiocyanate-conjugated anti-CD36, and respective control isotopes (mouse IgGs). A total of 5,000 cells were counted and the frequency of surface antigens was determined by FACscan. RESULTS: A significant positive link between LDL-cholesterol and the CD16+ subset ratio was found by linear correlation analysis (p < 0.05) but not by multivariate regression analysis. Both linear correlation analysis and ANOVA revealed a significant inverse link between high density lipoprotein (HDL)-cholesterol and the CD16+ subset ratio (both p < 0.01). By multivariate regression analysis, gender was the main significant determinant for the CD16+ subset ratio. When serum total cholesterol (TC) was excluded from the analysis to avoid the interference from collinearity between serum TC and LDL (r = 0.84), HDL-cholesterol became independently and inversely linked to the CD16+ subset ratio. There were independent inverse links between HDL-cholesterol and the counts of all monocytes, CD16+ monocytes, and CD36+ monocytes. CONCLUSIONS: Our results suggest that circulating HDL-cholesterol may be much more important than LDL-cholesterol in affecting the transformation of circulatory monocytes into macrophages. The inverse link between HDL-cholesterol and the number of macrophage precursors in peripheral blood might contribute partly to the well-known antiatherogenic effect of HDL-cholesterol. PMID- 11911033 TI - Presence of iceA1 but not cagA, cagC, cagE, cagF, cagN, cagT, or orf13 genes of Helicobacter pylori is associated with more severe gastric inflammation in Taiwanese. AB - BACKGROUND AND PURPOSE: The presence of the cag pathogenicity island (PAI) may enhance the virulence of Helicobacter pylori. This study determined the prevalence of genotypes of the PAI, IS605, and iceA1 in H. pylori strains in Taiwanese and whether these genotypes were related to the type of clinical disease or gastric pathology. METHODS: One hundred clinical isolates were collected from 33 duodenal ulcer, 41 nonulcer related dyspepsia, 14 gastric ulcer, and 12 gastric malignancy patients. Polymerase chain reaction (PCR) assays were performed for cagA, cagC, cagE, cagF, and cagN in the cagI region, cagT and orf13 in the cagII region, and IS605 and iceA1 in all H. pylori isolates. Gastric histology of the infected host was reviewed using the updated Sydney system. RESULTS: All strains were positive for all the selected genes in the PAI. PCR amplification found IS605 in 17%, while colony hybridization revealed it in 36% of strains. The prevalence of the cagA gene detected by PCR using cagA1, cagA2, and cagA3 primers was 26, 100, and 100%, respectively. The iceA1 gene existed in 72% of the H. pylori isolates. The mean ulcer size and the severity of acute gastric inflammation in patients infected with iceA1-positive strains were significantly greater than in those infected with iceA1-negative strains (p < 0.05). CONCLUSIONS: All clinical H. pylori isolates from different gastric diseases in our Taiwanese patients were positive for the PAI, but only 36% of these isolates carried an IS605 insertion. The selected genes in the PAI were not correlated with disease outcome. The determination of cagA prevalence is based on the selection of suitable primers. In contrast, bacterial factors such as the presence of iceAl could be related to severity of gastric inflammation and increase in ulcer size in H. pylori-infected patients. PMID- 11911035 TI - Antibiotic usage in community-acquired infections in hospitals in Taiwan. AB - BACKGROUND AND PURPOSE: Using an epidemiologically meaningful in-hospital population with community-acquired infections, we evaluated antibiotic therapy in terms of indication and choice of antibiotic and microbiologic work-up. METHODS: Infectious disease specialists evaluated charts of 436 patients from 9 hospitals and selected those who received antibiotics within 3 days of admission. Each antibiotic prescribed was marked for appropriateness of indication and choice. Microbiologic isolates were evaluated for their clinical significance. RESULTS: The most common infections were in the lower respiratory tract (46.1%). Each patient received a mean of 2.25 antibiotics for 8.1 +/- 6.4 days. Of the 975 courses of antibiotics given in the study period, indication and choice were correct in 37.4% and unsatisfactory in 14.5%. The vast majority of antibiotics used (79.2%) were first-line antibiotics--usually first-generation cephalosporins, aminoglycosides, and aminopenicillins. Most patients (66%) had a microbiology laboratory work-up, but only 37.4% were judged by evaluators to have a meaningful microbiologic diagnosis. Among the 201 patients with lower respiratory tract infections, 105 (52.2%) had a diagnosis of pneumonia. A positive isolate was recovered in 30 (28.6%) patients, and most of these isolates (20, 68.7%) were aerobic gram-negative rods. There were three positive blood cultures but none grew Streptococcus pneumoniae. CONCLUSIONS: Antibiotics were used excessively in number and duration. The microbiologic work-up had little effect on the indication and choice of antibiotics. Community-acquired pneumonia differed markedly from that in Western countries in that only 3.3% were caused by S. pneumoniae. PMID- 11911034 TI - Estrogen blocks parathyroid hormone-stimulated osteoclast-like cell formation in modulating differentiation of mouse marrow stromal cells in vitro. AB - BACKGROUND AND PURPOSE: During skeletal development and bone remodeling, bone marrow stromal cells give rise to osteoblasts and provide a critical microenvironment to support osteoclast formation. Estrogen is important for the maintenance of bone balance in adult animals by either increasing bone mass or inhibiting osteoclastic bone resorption. This study sought to determine the role that estrogen plays in coordinating osteogenic differentiation of bone marrow stromal cells and the ability of these cells to support osteoclast formation. METHODS: A conditionally immortalized mouse bone marrow stromal cell line, MS1, was used to examine the effects of estrogen on stromal cell differentiation and on stromal cell-supported osteoclast formation. RESULTS: On treatment of MS1 cells with 17 beta-estradiol (E2) (10(-12)-10(-8) M), alkaline phosphatase activity and bone nodule formation were increased in a dose-dependent manner, while the proliferation of MS1 cells was dose-dependently inhibited. 17 beta-E2 (10(-12)-10(-8) M) also caused a concentration-dependent inhibition of tartrate resistant acid phosphatase (TRAP)-positive multinucleated cell (MNC) formation in parathyroid hormone (PTH)-stimulated MS1 and spleen cell cocultures. Furthermore, estrogen pretreatment of MS1 cells also decreased the number of TRAP-positive MNCs in cocultures. Culturing in PTH-treated conditioned media did not rescue the loss of activity supporting osteoclast-like cell formation in MS1 cells. CONCLUSION: These results indicate that 17 beta-E2-stimulated osteoblastic differentiation of mouse marrow stromal cells results in bone matrix mineralization and a decrease in activity of supporting PTH-induced osteoclast like cell formation. PMID- 11911036 TI - Neonatal choroid plexus cysts and early childhood developmental outcome. AB - BACKGROUND AND PURPOSE: Choroid plexus cysts (CPCs) are incidental findings on sonograms of the neonatal head. The incidence of CPCs and their association with childhood neurodevelopmental outcome remains unclear. The purpose of this study was to determine the incidence of neonatal CPCs and their association with early childhood neurodevelopmental outcome. METHODS: Between July 1997 and June 1998, routine brain sonographic examinations were performed on 2,111 normal newborns. All neonates in whom CPCs were detected were followed up with serial sonograms at 1, 2, 4, and 6 months of age. Developmental milestones were subsequently evaluated at follow-up, with assessments performed according to the Denver II Developmental Screening Test. RESULTS: CPCs were identified in 186 neonates (8.8%), 14 (7.5%) of which were bilateral. The mean (+/- standard deviation) cyst diameter was 2.6 +/- 0.8 mm (range, 1.1-8.6 mm), with five cysts of more than 5 mm. Physical examinations were otherwise normal for all 186 neonates with a CPC. Follow-up ultrasonographic studies were completed for 155 children, with cysts regressing spontaneously in 137. By 6 months of age, residual cysts were visible in only 18 children (11.6%). Developmental outcome was normal for all 179 children who completed the scheduled follow-up ranging from 30 to 42 months (mean, 35 mo). CONCLUSIONS: CPCs were detected in 8.8% of neonates. Most of the cysts resolved spontaneously. The existence of isolated CPCs in the newborn was not associated with abnormal physical findings or with any delay in early childhood development. PMID- 11911037 TI - One-stage correction of proximal hypospadias and penoscrotal transposition. AB - BACKGROUND AND PURPOSE: Total correction of proximal hypospadias and penoscrotal transposition (PST) is a challenge to surgeons. Staged operation is usually recommended because the blood supply to the neourethra or the skin covering the penile shaft may be severed during scrotoplasty. This paper describes results obtained using a new technique for total correction, which preserves the blood supply to the neourethra in a one-stage operation. PATIENTS AND METHODS: Between July 1998 and March 2000, five boys (mean age 4 yr) with proximal hypospadias and PST underwent total correction in a one-stage operation. The urethral meatus of these patients was located at mid shaft in one, at the penoscrotal junction in two, and at the scrotum in two. Hypospadias was repaired using the Snodgrass procedure and PST was corrected using the Ehrlich and Scardino technique. Radical bulbar urethra dissection and tunica albugineal plication were used to correct penile curvature in all five cases. The urethral stent was removed on the seventh or eighth postoperative day. The meatus was then dilated using the cone tip of an ophthalmic ointment tube two or three times per day for 2 to 4 weeks. Postoperative urinary flow was observed in the outpatient clinic. RESULTS: The mean follow-up period was 11.2 months. There was no postoperative fistula. One patient had postoperative meatal stenosis that was successfully treated by dilation. Postoperatively, the penile base was well above the scrotal rhugae and the meatus was at the tip of the glans in each patient. The postoperative urinary flow was straight in all patients. CONCLUSION: Combining Snodgrass hypospadias repair and Ehrlich and Scardino PST repair in a one-stage operation preserved the blood supply to the neourethra and achieved excellent functional and cosmetic results. PMID- 11911038 TI - Association of changes in the pattern of urinary calculi in Taiwanese with diet habit change between 1956 and 1999. AB - OBJECTIVE: Due to rapid economic development over the past four decades, urinary stone components may have changed in Taiwan. We studied the changes in urinary stone components over time and the possible association with dietary changes during the same period. MATERIALS AND METHODS: From 1956 to 1999, 9,715 urinary calculi were collected at a single institution and analyzed using polarizing microscopy. Dietary information was obtained from an official national report. Linear regression was used to analyze the possible correlation between the change in stone components with daily consumption of animal protein, vegetable protein, and lipid. RESULTS: Eleven distinct components were identified. Calcium oxalate (Jensen type I stone; found in sterile, acidic urine) was found most frequently (87.3%), and its incidence increased gradually with time. However, the incidence of Jensen type III stone (caused by metabolic abnormality) gradually decreased from 1956 to 1999. The male to female ratio among subjects was 2.3:1, and the modal age was in the forties. Female patients were more likely to suffer from type II stones (found in infected, alkaline urine), whereas type I and III stones were more prevalent in males. Among the dietary components, consumption of animal and vegetable proteins and lipid increased significantly during the same period, and appeared to be coincident with the increased incidence of type I stones during the study period. CONCLUSIONS: Patterns of urinary tract stones in Taiwan have progressively changed in the past four decades and are now similar to those in western populations. The incidence of type I stones has increased during the past four decades, which may reflect the Westernization of dietary habits in Taiwanese during the same period. PMID- 11911039 TI - Healthcare utilization patterns and risk adjustment under Taiwan's National Health Insurance system. AB - BACKGROUND AND PURPOSE: Some recent proposals for reform of Taiwan's National Health Insurance (NHI) system include risk adjustment mechanisms. However, there is a paucity of research on risk adjustment and its utilization in health insurance systems in Taiwan. The purposes of this study were to determine the healthcare utilization pattern and to develop a risk assessment model for capitation payments under NHI. METHODS: The individual enrollment and medical expenditure data for 1996 and 1997 were obtained from the Bureau of NHI. A random sample of 360,037 beneficiaries was divided into two sub-samples: one for model building and one for validation. Linear regression was employed to estimate the relationship between each individual's 1997 total expenditure and risk adjusters, i.e., age, gender, prior years' medical spending, and catastrophic status. RESULTS: The 10- to 14-years age group had the lowest total expenditure of $NT 3,055 ($US 1 = $NT 27.5 in 1996) in 1996, while the 65 years and over age group had the highest at almost 10 times more than the lowest. The distributions of total expenditure for both genders followed the familiar J-shaped curve. The average of the total expenditure of individuals with a catastrophic diagnosis was more than 17 times that of individuals without. Age and gender resulted in a predictive R2 of only 3.8% in the risk assessment model. By including prior total expenditure, the predictive R2 increased to 24.2%. Further addition of catastrophic status increased the predictability slightly to 25%. Prior outpatient expenditure predicted 72% of subsequent outpatient expenditure, but prior inpatient expenditure predicted only 3% of subsequent inpatient expenditure. CONCLUSIONS: As in other countries, age and gender provided only limited predictability in risk assessment. On the other hand, prior outpatient expenditure in this study provided relatively superior predictability in risk assessment. Prudence is required when including prior utilization as a part of the risk assessment model in calculating capitation payments, as this may indirectly encourage unnecessary use of healthcare services. PMID- 11911040 TI - Factors influencing the long-term effects of supervised cardiac rehabilitation on the exercise capacity of patients with acute myocardial infarction. AB - BACKGROUND AND PURPOSE: Cardiac rehabilitation (CR) programs can effectively increase exercise capacity after an acute myocardial infarction (AMI). The purpose of this study was to investigate the changes in exercise capacity at the end of a supervised CR program and to determine the factors influencing exercise capacity 6 months after the end of a supervised CR program. METHODS: One hundred and four AMI patients who completed an 8-week supervised CR program were included in the study. Those who had an increased exercise capacity at the end of the 8 week supervised CR program were followed for 6 months to determine the possible factors influencing exercise capacity. Student's-t, chi-square, and ANOVA tests were used to make comparisons between groups and among stages of the program. RESULTS: All participants had a significant increase in exercise capacity and high-density lipoprotein cholesterol (HDL-C) at the end of the 8-week supervised CR program. The anaerobic threshold increased from 0.94 +/- 0.19 to 1.11 +/- 0.19 W/kg (p < 0.05) and serum HDL-C increased from 36.0 +/- 8.7 to 41.1 +/- 10.7 mg/dL (p < 0.05). Follow-up results demonstrated that smoking cessation, decrease in body mass index, and participation in leisure time physical activities were positively associated with increased exercise capacity. CONCLUSION: This study demonstrated that a supervised CR program improves exercise capacity and serum lipid profile in patients after AMI. Status of smoking, body mass index, and leisure time physical activities affect the long-term results of exercise training. PMID- 11911042 TI - Natural killer cell deficiency associated with Hodgkin's lymphoma: a case report. AB - Natural killer (NK) cells are large granular lymphocytes that play important roles in immunity against viral infection. NK cell deficiency is associated with recurrent episodes of severe herpes group virus reactivation. Few cases of NK cell deficiency have been reported. Here, we report the case of a Taiwanese girl who had suffered from severe atopic dermatitis since infancy, and recurrent episodes of herpes virus reactivation since the age of 3 years old. NK cell deficiency was diagnosed based on the finding of persistently low NK cell counts in peripheral blood. Hodgkin's lymphoma developed when she was 6 years old. The present case suggests that NK cell deficiency may be an important risk factor in the development of Hodgkin's lymphoma. PMID- 11911043 TI - Technical considerations in hand-assisted laparoscopic live donor nephrectomy: initial Taiwan experience from National Taiwan University Hospital. AB - This report describes the initial experience of laparoscopic live donor nephrectomy (lap-LDN) in Taiwan and discusses the technical considerations and modifications of our technique. From September to November 2000, three (one right and two left) lap-LDNs were performed at our institute. The right kidney was retrieved in one donor because of an early branching of the left renal artery. The details of our technique are described for both left and right LDN. The perioperative parameters were compared to those of the 10 immediately preceding cases of LDNs using the traditional open approach. All lap-LDNs and open LDNs were successful, and all 13 recipients had smooth recovery of renal function. The donors recuperated better in the lap-LDN group with resumption of oral intake on postoperative day (POD) 1 and discharge on POD 5 (vs POD 3.4 and 8.5, respectively, in the open group). The mean blood loss was lower and narcotic use was less in the lap-LDN group (75 vs 164 mL, 25 vs 47 mg morphine sulfate equivalent, respectively). The extraction wound was much shorter in the lap-LDN group (6.5-8 vs 23 cm). The warm ischemia time was slightly but not significantly shorter in the open group (4 vs 2.75 min), and the average operative time was shorter in the open group. The results of our initial experience suggest that for surgeons with laparoscopic surgery experience, lap-LDN is a feasible procedure that decreases donor discomfort, while improving the quality of graft kidneys and the safety of the donor. PMID- 11911041 TI - Hypopituitarism and nesidioblastosis in an elderly patient with orbital lymphoma. AB - Nesidioblastosis is a rare cause of hyperinsulinemic hypoglycemia in adults. We report a case of combined hypopituitarism with secondary adrenal insufficiency and nesidioblastosis with symptomatic hypoglycemia. This 84-year-old woman had a diagnosis of right side orbital lymphoma and underwent one complete course of local radiotherapy 6 months prior to this admission. Intermittent consciousness alteration had occurred in the 2 months after radiotherapy, and hypoglycemia with a blood glucose of 0.61 mmol/L (11 mg/dL) was noted. Although this case exhibited inappropriate hyperinsulinemia during the hypoglycemic episode, the prolonged fasting test was negative, which was unusual for insulinoma but common to nesidioblastosis. The hypopituitarism and secondary adrenal insufficiency, which may have been radiotherapy related, and the nesidioblastosis led to a relapse of neuroglycopenia. After a glucocorticoid supplement and an 80% subtotal pancreatectomy, her hypoglycemic symptoms were relieved. This case reminds us that nesidioblastosis, in addition to insulinoma, should be considered as a cause of hypoglycemia in elderly patients. PMID- 11911044 TI - Effects of experience and training on diagnostic accuracy. AB - In this reexamination of a previously published report (R. Brammer, 1997), psychologists and psychology students (N = 138) were provided an artificial intelligence program that simulated a clinical interview, The "client" provided paragraph-length answers to the questions participants chose to ask. At the end of their interview, the participants provided a brief diagnosis for the client. A path analysis revealed that clinical experience is a strong predictor of the ability of form an accurate diagnosis and that an individual's level of training, mediated by the number of diagnostic questions asked, also helps to derive accurate diagnoses. PMID- 11911045 TI - Underreporting of psychopathology on the MMPI-2: a meta-analytic review. AB - Meta-analytic techniques were applied to studies of the MMPI-2 in which participants given standard instructions were compared with participants instructed or believed to have been underreporting. Traditional and supplementary indices of underreporting yielded a mean effect size of 1.25, suggesting that underreporting respondents differ from those responding honestly by a little more than 1 standard deviation, on the average, on these scales. Analyses of classification accuracy suggested that several scales are moderately effective in detecting underreporting, although accuracy decreases if participants have been coached about validity scales. Base rates of defensive responding in relevant populations are reviewed, and methodological issues, including research designs, coaching, and incremental validity of supplementary underreporting scales, are discussed. PMID- 11911046 TI - Identifying persistently antisocial offenders using the Hare Psychopathy Checklist and DSM antisocial personality disorder criteria. AB - Early starting, lifetime criminal persistence has been called sociopathy, antisocial personality disorder, and psychopathy. There is, however, disagreement about its core features and which measure is best for identifying such individuals. In the 1st of 2 studies of male offenders (n = 74), we found a large association between scores on the Psychopathy Checklist-Revised (PCL-R; R. D. Hare, 1991) and the Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) antisocial personality disorder criteria scored as a scale. The second study (n = 684) replicated this finding and found that, as previously shown for PCL-R scores, a discrete class (or taxon) also underlies scores on items reflecting antisocial personality disorder. The high association among these sets of items and their similarity in predicting violence suggested that the same natural class underlies each. Results indicated that life-course-persistent antisociality can be assessed well by measures of psychopathy and antisocial personality disorder. PMID- 11911047 TI - A comparison of MMPI-2 validity in African American and Caucasian psychiatric inpatients. AB - This study investigated ethnic differences on the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) in 229 African American and 1,558 Caucasian psychiatric inpatients. Mean differences were found on several MMPI-2 validity and clinical scales. These were generally consistent with differences between the groups, indicated by the available extratest criterion data. To identify potential bias, the authors conducted 65 step-down hierarchical multiple regression analyses, predicting conceptually relevant clinical criteria from either MMPI-2 clinical or content scales for each gender. A number of MMPI-2 scales evidenced bias reflecting minor underprediction of psychopathology in African Americans. It is important to note that, in almost all cases, the magnitude of these differences was small and not clinically significant. PMID- 11911048 TI - Correspondence of psychiatric patient and informant ratings of personality traits, temperament, and interpersonal problems. AB - Psychological assessment of psychiatric patients frequently relies on self report, yet descriptions from patients often are regarded as suspect. Investigation of agreement between reports from patients versus knowledgeable informants is critical to assessing the validity of self-ratings. Self- and informant reports of temperament, personality traits, and interpersonal problems were collected from an adult, nonpsychotic psychiatric sample (N = 90). The majority of patients had depressive diagnoses (62%), were female (81%), and were Caucasian (98%). Few mean-level differences between self- and informant reports were found. Self-informant agreement correlations were comparable in magnitude and variability to findings from nonclinical samples. Results suggest that the overall effect of psychopathology on self-ratings of personality traits, temperament, and interpersonal problems was minimal in the authors' patient sample. This conclusion runs counter to the intuitively appealing notion that psychopathology has a detrimental effect on self-awareness. PMID- 11911049 TI - Differential item functioning in a Spanish translation of the PTSD checklist: detection and evaluation of impact. AB - This study demonstrated the application of an innovative item response theory (IRT) based approach to evaluating measurement equivalence, comparing a newly developed Spanish version of the Posttraumatic Stress Disorder Checklist-Civilian Version (PCL-C) with the established English version. Basic principles and practical issues faced in the application of IRT methods for instrument evaluation are discussed. Data were derived from a study of the mental health consequences of community violence in both Spanish speakers (n = 102) and English speakers (n = 284). Results of differential item functioning (DIF) analyses revealed that the 2 versions were not fully equivalent on an item-by-item basis in that 6 of the 17 items displayed uniform DIF. No bias was observed, however, at the composite PCL-C scale score, indicating that the 2 language versions can be combined for scale-level analyses. PMID- 11911050 TI - Multidimensional latent-construct analysis of children's social information processing patterns: correlations with aggressive behavior problems. AB - Social information processing (SIP) patterns were conceptualized in orthogonal domains of process and context and measured through responses to hypothetical vignettes in a stratified sample of 387 children (50% boys; 49% minority) from 4 geographical sites followed from kindergarten through 3rd grade. Multidimensional, latent-construct, confirmatory factor analyses supported the within-construct internal consistency, cross-construct discrimination, and multidimensionality of SIP patterns. Contrasts among nested structural equation models indicated that SIP constructs significantly predicted children's aggressive behavior problems as measured by later teacher reports. The findings support the multidimensional construct validity of children's social cognitive patterns and the relevance of SIP patterns in children's aggressive behavior problems. PMID- 11911051 TI - Identifying victims of peer aggression from early to middle childhood: analysis of cross-informant data for concordance, estimation of relational adjustment, prevalence of victimization, and characteristics of identified victims. AB - Two studies were conducted to investigate cross-informant measures of children's peer victimization. In Study 1, self- and peer reports of victimization were compared for 197 children from Kindergarten (M age = 5.73) to Grade 4. Before Grade 2, peer reports were less reliable than self-reports and were poor estimators of relational adjustment. In Study 2, single- versus multiple informant (self, peer, teacher, parent) victimization measures were compared for 392 children across grades 2 (M age = 8.73) to 4. Results indicated that (a) data from the four informants were reliable and increasingly concordant over time, (b) no single-informant measure proved to be the best predictor of relational adjustment, and (c) a multi-informant composite measure yielded better estimates of relational adjustment than any single-informant measure. PMID- 11911052 TI - The reliability and validity of the psychopathy checklist: youth version (PCL:YV) in nonincarcerated adolescent males. AB - Current knowledge about the validity of the psychopathy syndrome in youth is limited largely to studies relying on parent-teacher rating scales or slight modifications of adult measures. Recently, the Psychopathy Checklist: Youth Version (PCL:YV) was designed for use with adolescents. However, most studies that have used this measure examined incarcerated males and addressed only validity criteria related to antisocial behavior. We investigated the generality and construct validity of the psychopathy syndrome in an adolescent sample by assessing 115 adolescent males on probation with the PCL:YV. Reliability of measurement was high. PCL:YV ratings predicted not only antisocial behaviour but also other indices of childhood psychopathology, interpersonal behaviors associated with adult psychopathy, and a lack of attachment to parents. These findings suggest that the PCL:YV identifies a syndrome in adolescence consistent with theory and research on adult males. PMID- 11911053 TI - Leadership and patriotism. PMID- 11911054 TI - Compliance delay for HIPAA transaction regulations. PMID- 11911055 TI - Implant placement in the vertically enhanced ridge--a surgical and restorative interdisciplinary approach. AB - A diagnostic and therapeutic approach is offered to enable the surgeon and restorative dentist to successfully install endosseous implants into the vertically enhanced ridge. The strategy involves the use of diagnostic and surgical templates to aid in the creation of positive osseous architecture followed by the vertical orientation of the endosseous implant(s). This effort provides optimal hard-tissue and soft-tissue form for the creation of the most favorable crown profiles for implant-supported prostheses. PMID- 11911056 TI - Endodontics in the 21st century: the rotary revolution. AB - Endodontic therapy is becoming an increasingly routine facet of general dental practice. Advances in technology and techniques have made endodontics in the contemporary clinical environment more pleasant and predictable for both the patient and the provider. The development of rotary instrumentation with nickel titanium files has revolutionized the cleaning and shaping aspects of treatment. A sequenced model is discussed that has been shown to have encouraging outcomes in both predoctoral and continuing education experiences. The protocol may use more instruments than some other approaches, but has evolved as a technique that provides reliable and predictable outcomes. PMID- 11911057 TI - Subcutaneous emphysema after restorative dental treatment. AB - Subcutaneous emphysema is an uncommon phenomenon in dentistry, usually occurring with the use of air-driven, high-speed handpieces during dental and oral surgery, operative, endodontic, or periodontal treatment. Air is forced into a surgical wound or subepithelial laceration in the oral cavity, dissecting through the different layers of tissue fasciae, and usually creating a unilateral enlargement of the facial and/or submandibular regions. This occurs with or without crepitus, pain, and airway obstruction. Treatment usually consists of antibiotic and mild analgesic therapy, close observation, and reassurance by the attending dentist. Symptoms generally subside in 3 to 10 days; however, consultation with a physician is necessary to rule out further complications. PMID- 11911058 TI - The all-ceramic cantilever bridge: a variation on a theme. AB - All-ceramic crowns have become a dependable treatment modality. As a result, there is interest in expanding this modality to all-ceramic bridge-work. This article describes a modification of the CAD/CAM all-ceramic Procera technique, which allows the fabrication of a single-unit cantilever ceramic bridge using existing equipment and procedures. The stress-bearing areas of the bridge are made of densely sintered high-purity alumina without ceramic-fusing material. Although the indications for this bridge are limited, this technique adds to the dentist's armamentarium of esthetic restorations. PMID- 11911059 TI - Casting alloy trends in restorative dentistry. PMID- 11911060 TI - The immunocompromised health care professional. PMID- 11911061 TI - Replacement of an anterior tooth with a fiber-reinforced resin bridge. PMID- 11911062 TI - The basics of practice management. PMID- 11911063 TI - New technologies in adult orthodontics. Interview by Mark J. Friedman. PMID- 11911064 TI - Primary vascular-type craniofacial pain. AB - Primary vascular-type craniofacial pain comprises a group of pain disorders that share common diagnostic features. These are unilateral, episodic, pulsatile, severe pain. Accompanying phenomena include local autonomic (e.g., tearing, rhinorrhea) and systemic signs (e.g., nausea, photophobia). Primary vascular-type craniofacial pain includes migraine, cluster headache, and paroxysmal hemicrania. A new diagnostic entity, vascular orofacial pain, is suggested. Treatment of primary vascular-type craniofacial pain depends on its more specific diagnosis, and may be abortive or prophylactic. Diagnostic features, common pathophysiological mechanisms, and treatment modalities are discussed. PMID- 11911065 TI - Quality control of the automatic processor. AB - An often ignored component of a radiology quality control program is daily sensitometric testing of automatic film processors. Sensitometric strip tests can reveal fluctuating chemical activity levels of developing solutions that can have a detrimental effect on developing films. By performing a daily sensitometric strip test before patient films are developed, time, cost, and patient radiation exposure are minimized, which reduces the number of retakes as a result of processing problems. Sensitometric strip tests also assist in quality control by making troubleshooting more specific. PMID- 11911066 TI - Training pathways for careers in dental, oral, and craniofacial research. AB - There are virtually unlimited opportunities for biomedical research training through either extramural or intramural training mechanisms, with programs designed for both the pre- and postdoctoral educational levels. The only difficulty is selecting from the vast array of research topic areas. PMID- 11911068 TI - Management systems and software. AB - To ensure that your software optimizes your practice management systems, design systems that allow you and your team to achieve your goals and provide high levels of quality dentistry and customer service to your patients. Then use your current software system or purchase a new practice management software program that will allow your practice to operate within the guidelines of the systems which you have established. You can be certain that taking these steps will allow you to practice dentistry with maximum profitability and minimum stress for the remainder of your career. PMID- 11911067 TI - Optimal esthetic results with indirect posterior composite resins. PMID- 11911069 TI - Computer-assisted analysis of the oral brush biopsy. AB - Computer-assisted analysis of the oral brush biopsy is a recently introduced tool that determines the significance of an oral lesion. The oral brush biopsy is minimally invasive, requires no anesthesia, and definitively distinguishes benign from precancerous and cancerous lesions. Oral brush biopsy specimens are analyzed with the aid of a highly specialized neural network-based computer system specifically designed to detect oral epithelial precancerous and cancerous cells. PMID- 11911071 TI - Introducing esogetic colorpuncture: a wholistic system of acu-light therapy for body, mind & soul. PMID- 11911070 TI - The Florence Nightingale Moment: May 12, 2002. Nurture our nurses around the world. PMID- 11911072 TI - Environmentally safer traveling. Part I: Automobiles. PMID- 11911073 TI - Regulation of the ion-transporting mitochondrion-rich cell during adaptation of teleost fishes to different salinities. AB - The mitochondrion-rich cells (MRCs) in teleost gill and equivalent tissues are important osmoregulatory sites in maintaining ionic balance. These cells express a variety of ion pumps, transporters, and channels, which play central roles in ionic regulation. Recently, two types of MRCs have been identified in euryhaline fishes: seawater (SW)-type MRCs extrude Na and Cl ions in SW conditions; freshwater (FW)-type MRCs take up at least Cl-. Long-term development/differentiation of the two types of MRCs during adaptation to different salinities appears to be regulated mainly by endocrine factors. Osmolality, Ca2+, neurotransmitters, and fast-acting hormones rapidly regulate the SW MRCs. Recent information is assembled in this review and suggests the functional plasticity of highly specialized MRCs. PMID- 11911074 TI - The crustacean eye: dark/light adaptation, polarization sensitivity, flicker fusion frequency, and photoreceptor damage. AB - Compound eyes, nauplius eyes, frontal organs, intracerebral ocelli, and caudal photoreceptors are the main light and darkness detectors in crustaceans, but they need not be present all at once in an individual and in some crustaceans no photoreceptors whatsoever are known. Compound eye designs reflect on their functions and have evolved to allow the eye to operate optimally under a variety of environmental conditions. Dark-light-adaptational changes manifest themselves in pigment granule translocations, cell movements, and optical adjustments which fine-tune an eye's performance to rapid and unpredictable fluctuations in ambient light intensities as well as to the slower and predictable light level changes associated with day and night oscillations. Recycling of photoreceptive membrane and light-induced membrane collapse are superficially similar events that involve the transduction cascade, intracellular calcium, and membrane fatty acid composition, but which differ in aetiology and longterm consequence. Responses to intermittant illumination and linearly polarized light evoke in the eye of many crustaceans characteristic responses that appear to be attuned to each species' special needs. How the visual responses are processed more centrally and to what extent a crustacean makes behavioural use of e-vector discrimination and flickering lights are questions, however, that still have not been satisfactorily answered for the vast majority of all crustacean species. The degree of light induced photoreceptor damage depends on a large number of variables, but once manifest, it tends to be progressive and irreversible. Concomittant temperature stress aggravates the situation and there is evidence that free radicals and lipid hydroperoxides are involved. PMID- 11911075 TI - Vacuolar-type H(+)-ATPase and Na+, K(+)-ATPase expression in gills of Atlantic salmon (Salmo salar) during isolated and combined exposure to hyperoxia and hypercapnia in fresh water. AB - Changes in branchial vacuolar-type H(+)-ATPase B-subunit mRNA and Na+, K(+) ATPase alpha- and beta-subunit mRNA and ATP hydrolytic activity were examined in smolting Atlantic salmon exposed to hyperoxic and/or hypercapnic fresh water. Pre smolts, smolts, and post-smolts were exposed for 1 to 4 days to hyperoxia (100% O2) and/or hypercapnia (2% CO2). Exposure to hypercapnic water for 4 days consistently decreased gill vacuolar-type H(+)-ATPase B-subunit mRNA levels. Salmon exposed to hyperoxia had either decreased or unchanged levels of gill B subunit mRNA. Combined hyperoxia + hypercapnia decreased B-subunit mRNA levels, although not to the same degree as hypercapnic treatment alone. Hyperoxia generally increased Na+, K(+)-ATPase alpha- and beta-subunit mRNA levels, whereas hypercapnia reduced mRNA levels in presmolts (beta) and smolts (alpha and beta). Despite these changes in mRNA levels, whole tissue Na+, K(+)-ATPase activity was generally unaffected by the experimental treatments. We suggest that the reduced expression of branchial vacuolar-type H(+)-ATPase B-subunit mRNA observed during internal hypercapnic acidosis may lead to reduction of functional V-type H(+) ATPase abundance as a compensatory response in order to minimise intracellular HCO3- formation in epithelial cells. PMID- 11911076 TI - Possible involvement of nitric oxide in signaling pigment dispersion in teleostean melanophores. AB - The possible involvement of nitric oxide (NO) in regulating the motile activities of teleostean melanophores was studied in the dark chub Zacco temmincki (Cyprinidae, Cypriniformes) and in the translucent glass catfish Kryptopterus bicirrhis (Siluridae, Siluriformes). NO donors, including (+/-)-(E)-methyl-2-[(E) hydroxyimino]-5-nitro-6-methoxy-3-hexaneamide (NOR1), molsidomine (MSD), sodium nitroprusside (SNP) and glyceryl trinitrate (GTN), had no pigment-aggregating action on melanophores, but actively dispersed melanosomes in those cells. Among those reagents, NOR 1, a spontaneous releaser of NO, was the most effective. Inhibitors for nitric oxide synthase (NOS), i.e. N omega-nitro-L-arginine methyl ester (L-NNA), N omega-nitro-L-arginine (L-NAME) and N omega-monomethyl-L arginine (L-NMMA), showed melanosome-aggregating effects. A membrane-permeable analogue of cyclic guanosine-3',5'-monophosphate (8-Br-cGMP) was effective in dispersing melanosomes. The sum of these results suggests that NO plays an active role in the elaborate control of color changes in teleosts by dispersing pigment in melanophores via activation of soluble guanylyl cyclase to increase cytosolic levels of cGMP. PMID- 11911077 TI - Manipulation of the somatosensory cortex modulates stimulus-induced repetitive ear movements in a seizure-sensitive strain of gerbil. AB - Some Mongolian gerbils (Meriones unguiculatus) respond to stimulation by seizures, the pattern of which changes progressively during development. We previously established a seizure-sensitive strain, MGS/Idr, in which all animals exhibit such stimulus-induced seizures. We have now noted that all adults of this strain also show repetitive backward movements of the ears at the ears at the beginning of stimulus-induced seizures, although the incidence varies with the individual. We examined whether the cerebral cortex was involved in these movements and found that electrical stimulation of an area of the somatosensory cortex elicited strong backward movement of the ear on the contralateral side, and that unilateral application of bicuculline, a GABAA receptor antagonist, induced spontaneous repetitive backward movements of the same ear. In this area, sharp waves appeared in the electrocortigram during the repetitive ear movements induced by seizure-inducing stimuli. Unilateral ablation of this area abolished stimulus-induced repetitive movements of the contralateral ear, but had no effects on those of the ipsilateral ear. These results suggest that, in certain types of seizure-susceptible subjects, it may be possible to modify stimulus induced repetitive movements by manipulating a certain area of the somatosensory cortex which is related to these movements and that this gerbil strain may be useful in research on this subject. PMID- 11911078 TI - Inhibitory effect of postpartum lesions or cuts in median raphe nucleus on maternal behavior in female rats. AB - In order to clarify the role of the median (MRN) and dorsal (DRN) raphe nuclei in regulating maternal care (retrieving and licking behavior), radiofrequency lesions or microknife cuts were made in postpartum rats on the day after delivery. Animals were housed individually without pups after the operation. One week after the surgery, maternal behavioral test was carried out daily for 3 days using pups of 2-6 days age. The results demonstrated that rats with MRN lesions or ventral horizontal cuts of the MRN showed extremely low incidence of the maternal behavior, as compared to those in control and sham-operated groups. DRN lesions or dorsal cuts of the MRN had no effect. In locomotor activities measured by the infrared sensor system, there was no difference between the groups. This suggest that the MRN but not DRN plays a critical role in regulating retrieving and licking behaviors and ventral outputs are involved in this function in postpartum rats. PMID- 11911079 TI - EST analysis of genes that are expressed in the neural complex of Ciona intestinalis adults. AB - A subtractive cDNA library was made corresponding to mRNAs expressed in the neural complex relative to those expressed in the pharynx of adults of the ascidian Ciona intestinalis. Determination and comparison of expressed sequence tags (ESTs) of a set of 1,527 randomly selected clones demonstrated that they represent 832 independent sequences. Five hundred seventy-two of the clones contained amino-acid-encoding sequences. BLASTX analyses showed that 342 of the 572 clones were strong matches (P < 10(-7)) to previously identified proteins, while the remaining 230 fell into the "no match" category. Among the clones matching previously identified proteins, about 80 clones represented proteins that are involved in the formation, maintenance of the structure, and function of the nervous system: 22 proteins are associated with signal transduction, five proteins are related to the synapse, 11 to transcription factors, nine to transporters, five to enzymes, and 13 to extracellular matrix and cytoskeletal components, and six to apoptosis. In addition, sequence information for genes associated with the immune system and for genes encoding proteins with interesting functions were obtained. These data provide cues for further studies on genes that are expressed in and function in the ascidian nervous system. PMID- 11911080 TI - Novel structures in secreting the androgenic gland hormone. AB - The secretory granules in the androgenic gland of the terrestrial isopod Armadillidium vulgare, which have been indistinct for long time because of vulnerable structures, were revealed by using the rapid-freezing and freeze substitution method. The fine structure of the androgenic gland is conspicuous by the distribution of numerous particular organelles in the cytoplasm consisting of the endoplasmic reticulum and the Golgi complex, and by having a number of highly organized structures developed between the androgenic gland cells. The structures connect to the intercellular space, which is seen as intercellular canaliculi for exporting the androgenic gland hormone. The plasma membranes near the particular structure of the intercellular canaliculi in the androgenic gland are often specialized to form cellular junctions. The secretory granules including the electron-dense materials, which are supposed to be peptides of androgenic gland hormone, are distributed beside the particular structure of the intercellular canaliculi. Some of the granules are seen to fuse with the plasma membranes. This observation suggests that, in the Armadillidium vulgare, the secretory granules containing androgenic gland hormone are transferred to the extracellular space through the intercellular canaliculi particularly developed for exporting the peptide hormone. This is the first evidence to show the secretory mechanism of the androgenic gland hormone in the Isopoda. PMID- 11911081 TI - Involvement of calcium, inositol-1,4,5 trisphosphate and diacylglycerol in the prothoracicotropic hormone-stimulated ecdysteroid synthesis and secretion in the prothoracic glands of Bombyx mori. AB - The objective of this study was to determine which intracellular second messenger systems are activated by prothoracicotropic hormone in the prothoracic glands (PGs) of Bombyx mori. Recombinant prothoracicotropic hormone (rPTTH) could stimulate ecdysteroid synthesis and secretion from day 6 PGs of the 5th instar of Bombyx mori within 30 min of in vitro incubation. However, rPTTH did not stimulate any increases in the glandular content of inositol 1,4,5-trisphosphate and cAMP during this short incubation period. Extracellular Ca2+ influenced the basal and rPTTH-stimulated ecdysteroid synthesis and release in a dose-dependent manner. The L-type Ca2+ channel antagonist, nitrendipine, inhibited the rPTTH stimulated ecdysteroid synthesis and secretion (IC50-28 microM). The phospholipase C inhibitor, 2-nitro-4-carboxyphenyl-N, N-diphenylcarbamate, inhibited the rPTTH-stimulated ecdysteroid synthesis (IC50-19 microM). The protein kinase C inhibitor, chelerythrine chloride, inhibited the rPTTH stimulated ecdysteroid synthesis (IC50-14 microM). The protein kinase C activator, phorbol-12-myristate 13-acetate (PMA), could stimulate basal ecdysteroid synthesis and secretion (EC50-1 microM) and its inactive alpha-isomer (4 alpha-PMA) was ineffective. The combined results suggest that the PTTH stimulated ecdysteroid synthesis and release in the PGs of Bombyx is dependent on extracellular Ca2+ and the bifurcating second messenger signalling cascade of inositol 1,4,5-triphosphate and diacylglycerol. PMID- 11911082 TI - Relative potency of three homologous natriuretic peptides (ANP, CNP and VNP) in eel osmoregulation. AB - Evidence has accumulated that atrial natriuretic peptide (ANP) plays important roles in sea-water adaptation in eels. However, the roles of the other two natriuretic peptides (CNP and VNP) in osmoregulation have not been examined yet. In the present study, the effects of homologous ANP, CNP and VNP were compared on plasma Na+ concentration (an indicator of plasma osmolality), hematocrit (an approximate indicator of blood volume) and drinking rate in freshwater- and seawater-adapted eels. In seawater eels, ANP and VNP, but not CNP, infused at 5 pmol/kg/min decreased plasma Na+ concentration and drinking rate and increased hematocrit. In freshwater eels, ANP and VNP failed to decrease plasma Na+ concentration but increased hematocrit to the same extent as in seawater eels. Inhibition of drinking was not detectable in freshwater eels because of little drinking before NP infusions. These results show that the effects of NPs on plasma Na+ concentration, drinking rate and hematocrit are mediated by NPR-A, since only ANP and VNP that bind with higher affinity to NPR-A are effective in seawater eels. The mechanisms of regulation of plasma Na+ concentration and hematocrit are unknown, but NPR-A is present in the responsible tissues for regulation of hematocrit in both freshwater and seawater eels. However, NPR-A may be absent in the tissues of freshwater eels that are responsible for regulation of plasma Na+ concentration. PMID- 11911083 TI - Some nemerteans (Nemertea) from Queensland and the Great Barrier Reef, Australia. AB - Three species of marine nemerteans described and illustrated from Queensland and the Great Barrier Reef, Australia, include one new genus and two new species: these are the monostiliferous hoplonemerteans Thallasionemertes leucocephala gen. et sp. nov. and Correanemertes polyophthalma sp. nov. A new colour variety of the heteronemertean Micrura callima is also reported, this species previously only being known from Rottnest Island, Western Australia. A key for the field identification of the marine nemerteans recorded from coastal Queensland and the Great Barrier Reef is provided. PMID- 11911084 TI - Description of a new species of Thais (Mollusca: Neogastropoda: Muricidae) from Taiwan, based on morphological and allozyme analyses. AB - Thais keluo sp. nov. is described from intertidal shores of southwest Taiwan. The new species is differentiated from five other closely related species, namely T. bitubercularis (Lamarck), T. jubilaea Tan and Sigurdsson, T. clavigera (Kuster), T. luteostoma (Holten) and T. rufotincta Tan and Sigurdsson, all of which occur in the South China Sea, on the basis of shell, radula and penis morphology. Thais keluo is also distinguished from the latter three species based on allozyme electrophoresis. The shell of T. keluo is characterized by four raised, spiral bands on the last whorl, one or two small, oblique columellar plica(e) on the inner lip, a finely crenate, thin, narrow, reddish-brown outer lip edge and four white, papillate denticles inside the outer lip of the aperture. In males, the penis is curved with a long, simple flagellum. The UPGMA cluster analysis based on 9 enzyme loci revealed that T. luteostoma is more closely related to T. clavigera than to T. keluo n.sp. The Nei's genetic distance (D) obtained between the new species and T. clavigera/T. luteostoma was 0.31, while T. clavigera and T. luteostoma were separated by a distance of 0.16. Thais rufotincta was separated from the other species by a distance of 0.78. In contrast, phylogenetic analysis of morphological data by maximum parsimony suggested that T. luteostoma was more closely related to T. keluo than to T. clavigera. However, both analyses indicated the close relationship amongst T. clavigera, T. luteostoma and the new species in relation to T. rufotincta. PMID- 11911085 TI - A new distinctive species of pagurid hermit crab (Crustacea: Decapoda: Anomura) from Japan. AB - A new species of pagurid hermit crab, Pagurus decimbranchiae, is described and illustrated based on 20 specimens collected from shallow waters of the Pacific coast of Japan ranging from Boso Peninsula to Tanegashima Island. It is quite distinctive in having the rudimentary arthrobranch on the third maxilliped represented by a single bud, however close morphological similarity is found between the new species and P. moluccensis Haig and Ball. Comparisons are also made among other species, including P. boriaustraliensis Morgan, P. sp. cf. boriaustraliensis sensu Rahayu and Komai (2000) and the members of the P. anachoretus group. The present generic assignment of the new species should be considered provisional, as more extensive study is needed to investigate phylogenetic relationships of the new species and the other species of Pagurus. PMID- 11911086 TI - An evaluation of the use of two-dimensional gel electrophoresis in proteomics. AB - With whole genomes being sequenced almost routinely, the next logical step towards a better understanding of cellular mechanisms lies in studying the functional units of gene expression-proteins. One of the fundamental approaches in proteomics is the use of two-dimensional gel electrophoresis as a mode of separation and visualization of complex protein mixtures. Despite several limitations of the method, its ability to separate large numbers of proteins, including their post-translationally modified forms, ensures that it will continue to be popular in several well-defined areas of proteomics. In this article, we discuss the merits and drawbacks of two-dimensional gels and compare them with alternative systems such as one-dimensional gels and liquid chromatography-based separation methods. In the wake of recent advances in mass spectrometry and related areas, we outline areas where two-dimensional gels can best be utilized as the preferred separation method in proteomic strategies. PMID- 11911087 TI - Proteome, transcriptome and genome: top down or bottom up analysis? AB - Biological systems are comprised of protein components found at a wide variety of abundances from millions of molecules of a single species per cell to less than one copy per cell. Because of this wide range of concentrations, measurement or a full accounting of each system is presently unavailable. Conventional separation and analytical methods (two-dimensional gel electrophoresis and mass spectrometry) allow identification and quantitation of many of the most abundant gene products (top down methods); and the majority of gene products, which are found at low abundance, can be neither identified nor measured in complex mixtures at present. The gene products that are found at low levels can be characterized and their properties analyzed by preparing ordered gene libraries of limited complexity from mRNA. When such preparations are expressed in cell free systems and analyzed by two-dimensional gel electrophoresis, the features of the gene products are available for analysis. This 'bottom up' approach allows identification of gene product properties so that analytical procedures can be devised and applied to complex mixtures. PMID- 11911088 TI - Proteomics: posttranslational modifications, immune responses and current analytical tools. AB - The publication of the human genome sequence enables most of the still unknown protein sequences to be added to the current databases. A sequence alone does not, however, give information about the possible expression level of the corresponding protein, neither does it inform about the possible posttranslational modifications, like phosphorylation, glycosylation or changes in individual amino acids. Thus, the human proteome project, a large scale analysis of the functions of gene products, will have an enormous impact on our understanding of the biochemistry of proteins, processes and pathways they are involved in. The diversity in proteins is considerably expanded by various posttranslational modifications. These also pose problems to the investigators, but their careful analysis often pays back because they can reveal important properties in proteins or peptides--like an increased antigenicity leading to (auto)immune responses or an active form of a signaling protein. Immune tolerance usually exists towards self-proteins, but in specific cases it may be broken by posttranslational modifications in the proteins. Novel mass spectrometric, affinity and display techniques offer valuable tools for the large-scale analysis of proteomes. In the present paper we discuss their use for the detection of posttranslational modifications, functional interactions and possible disease associated abnormalities in proteins. PMID- 11911089 TI - The role of mass spectrometry in proteome studies. AB - Mass spectrometry (MS) is an important tool in modern protein chemistry. In proteome analyses the expression of hundreds or thousands of proteins can be monitored at the same time. First, complex protein mixtures are separated by two dimensional gel electrophoresis (2-DE) and then individual proteins are identified by using MS followed by database searches. Recent developments in this field have made it possible to do automated, high-throughput protein identification that is needed in proteome analyses. MS can also be used to characterize post-translational modifications in proteins and to study protein complexes. This review will introduce the current MS methods used in proteome studies, and discuss their advantages and disadvantages. New instrumental MS developments are also presented that are useful in these analyses. PMID- 11911090 TI - Analysis of N-linked oligosaccharides: progress towards the characterisation of glycoprotein-linked carbohydrates. AB - The covalent attachment of carbohydrate to proteins is a very common co- or post translational event in the biosynthesis of glycoproteins. The type and heterogeneity of these oligosaccharides can affect a range of physico-chemical and biological properties of a glycoprotein. Thus the development of sensitive, reliable and robust analytical methods for carbohydrate analysis is important in the pharmaceutical industry, especially in the recombinant production of experimental and therapeutic glycoproteins. In this report we have reviewed methodology for the in-gel enzymatic release of N-linked oligosaccharides from glycoproteins separated by electrophoresis. These oligosaccharides are derivatised by reductive amination using 3-acetamido-6-aminoacridine (AA-Ac), a novel, highly fluorescent probe. A major advantage of this technique is that glycan derivatives are amenable to analysis by an array of chromatographic and mass spectrometric methods, allowing the resolution and characterisation of a wide variety of glycan structures. It is hoped that in due course the methodology described will be applied to proteomics studies, especially in identifying the role of carbohydrate in protein function and disease. PMID- 11911091 TI - Bioinformatics in proteomics. AB - Several genome sequencing projects have recently been completed and the majority of human coding regions have been sequenced. In the next step many of the further studies will concentrate on proteins. Proteomics methods are essential for studying protein expression, activity, regulation and modifications. Bioinformatics is an integral part of proteomics research. The recent developments and applications in proteomics are discussed including mass spectrometry data analysis and interpretation, analysis and storage of the gel images to databases, gel comparison, and advanced methods to study e.g. protein co-expression, protein-protein interactions, as well as metabolic and cellular pathways. The significance of informatics in proteomics will gradually increase because of the advent of high-throughput methods relying on powerful data analysis. PMID- 11911092 TI - [Opioid therapy misuse in persistent somatoform pain disorder]. PMID- 11911093 TI - [Pain and right-sided hemiparesis]. PMID- 11911094 TI - Predictive value of endoscopic ultrasonography for regression of gastric low grade and high grade MALT lymphomas after eradication of Helicobacter pylori. PMID- 11911095 TI - Recurrence of biliary symptoms after endoscopic sphincterectomy for choledocholithiasis in patients with gallbladder stones. PMID- 11911096 TI - Position statement on defining scholarship for the discipline of nursing. AB - While the mission of institutions of higher learning is unique in each setting, the commitment to scholarly approaches to education, practice, and research creates common bonds across the academic nursing community. This document is intended to clarify, extend, and enhance the scholarly work of nursing in academic settings. The application of the standards proposed in this document will differ by institution, yet will provide a framework for the advancement of nursing knowledge that will ultimately improve the health of people. PMID- 11911097 TI - Pay per view: the Arizona telemedicine program's billing results. PMID- 11911098 TI - Child abuse prevention program with POTS-based telehealth: a feasibility project. PMID- 11911099 TI - High frame rate Doppler echocardiography in the rat: an evaluation of the method. AB - AIMS: In small animal models, two-dimensional (2D) and Doppler echocardiography should provide more information than M-mode, especially in animals with infarcted and distorted left ventricles, but has been limited by low frame rates and poor near field resolution. New, high frame rate echo-Doppler equipment with digital processing was tested for accuracy of measurements. METHODS AND RESULTS: Fourteen normal Wistar rats (232-328 g) were examined under halothane anaesthesia. Pulsed Doppler recordings from both left ventricular outflow tract(LVOT) and right ventricular outflow tract (RVOT) cor-responded well with simultaneous ultrasound transit time measurements of aortic flow (LVOT: v=0.99x+4.8, min R=0.93. Standard error of estimate (SEE)=8.3 ml x min(-1), and RVOT: v=0.97x -4.3. R=0.93. SEE =8.4 ml x min(-1). No systematic differences were observed over a flow range of 20-90 ml x min(-1). Left ventricular (LV) dimensions assessed by 2D parasternal long-axis and short-axis views were equal to M-mode measurements with LV diameter 6.6 + 0.44 mm, anterior wall 1.8 +/- 0.18 mm, and posterior wall 1.5 + 0.56 mm. Mean absolute difference 4.4-8.5%. Intra- and interobserver variability was 4.6 +/- 4.1% and 6.7 +/- 7.0% for Doppler measurements, and 4.3 +/- 3.8% and 3.8 +/- 4.6% for dimensions, respectively. CONCLUSION: High frame rate Doppler echocardiography provides accurate non-invasive measurements of cardiac structure and function in the rat. PMID- 11911100 TI - The contribution of hysterectomy to the occurrence of urge and stress urinary incontinence symptoms. AB - OBJECTIVE: To study the contribution of hysterectomy to the occurrence of urge-or stress urinary incontinence symptoms. DESIGN: A population-based, cross-sectional cohort study conducted in 1999. SETTING: A university medical centre in The Netherlands. POPULATION: Random sample of 2322 women, between 35 and 70 years of age, from a suburban area in the central part of The Netherlands. METHODS: Self report questionnaire with questions from the Urogenital Distress Inventory, sociodemographic variables and data on obstetric and gynaecological history. MAIN OUTCOME MEASURES: Multivariate logistic regression analysis was used to obtain odds ratios with 95% confidence interval of the type and bothersomeness of urinary incontinence for hysterectomy. RESULTS: One thousand, six hundred and twenty-six women (70%) responded. The adjusted odds of urge (1.9, 1.4;2.6) and bothersome urge (2.6, 1.4;4.4) urinary incontinence were increased for women who had a hysterectomy. This association was not limited to the elderly but also occurred in women under 60 years of age. No increased odds of stress or bothersome stress incontinence were found. CONCLUSIONS: Meta-analysis has shown that hysterectomy increases the odds of urinary incontinence by 30%. However, a distinction between the different types of urinary incontinence symptoms has not been made. Such a distinction is of importance since urge incontinence gives a significantly greater reduction in health-related quality of life as compared with stress incontinence. Therefore, our finding that women scheduled for hysterectomy have an increased risk of developing urge incontinence symptoms indicates that these women should be counselled about this particular consequence. PMID- 11911102 TI - Substandard care in immigrant versus indigenous maternal deaths in The Netherlands. AB - As part of the Confidential Enquiry into the Causes of Maternal Deaths in The Netherlands, substandard care was assessed in immigrant versus indigenous maternal deaths. Except for substandard care related to the women's and relatives' decisions, substandard care factors were hypothesised to occur in similar frequency among both groups of women. The results, however, indicate that substandard care factors related to all aspects of care were disproportionately more frequent in immigrant women. More research into the interpretation of these worrying data is needed. PMID- 11911101 TI - A double-blind, randomised, placebo controlled study on the influence of carbohydrate solution intake during labour. AB - OBJECTIVE: Although there has been much debate on whether women should be allowed to eat and drink during labour, little scientific data are available on the effects of caloric intake on the course of labour. DESIGN: Double-blind, randomised, placebo controlled. SETTING: Leyenburg Hospital, The Hague, The Netherlands. POPULATION: Two hundred and one consecutive nulliparous women, pregnant of a single fetus in cephalic presentation. METHODS: All women were included in early labour (2cm-4cm of cervical dilatation) and were allowed to drink at will. MAIN OUTCOME MEASURES: The duration of labour, the need for augmentation and pain medication and the incidence of abdominal and vaginal instrumental deliveries. RESULTS: Drinking of carbohydrate solutions was well tolerated, but did not show any beneficial effects regarding labour outcome when compared with the control group. In the carbohydrate group, a higher caesarean section rate was observed (RR 2.9, 95% CI 1.29-6.54). CONCLUSIONS: Women in the carbohydrate group had worse labour outcome. It is unclear whether a statistical coincidence, a negative effect of the carbohydrate intake or an incorrect carbohydrate intake strategy is responsible for these results. Further studies are necessary before any definite conclusion can be drawn. PMID- 11911103 TI - Alternatives to Kinevac: shortages lead to inventive measures. PMID- 11911104 TI - Three Nobelists who paved the way. PMID- 11911105 TI - Red cell aplasia due to erythrovirus parvovirus B19 infection in a Jehovah's witness with post-transplant EBV-induced aggressive lymphoma: management considerations. AB - The refusal of Jehovah's Witnesses to accept blood and blood products often poses a clinical dilemma to present day medicine. We present a case of a Jehovah's Witness who had undergone renal transplantation, only to develop an Epstein-Barr virus associated aggressive lymphoma post-transplant. His condition was further complicated by erythrovirus (parvovirus) B 19 infection resulting in red cell aplasia and severe anemia. The management of this difficult clinical situation is discussed together with a review of recommendations for chemotherapy treatment in Jehovah's Witnesses. PMID- 11911108 TI - Contact leukoderma caused by buspirone patches. PMID- 11911106 TI - Incidence of MLL rearrangement in acute myeloid leukemia, and a CALM-AF10 fusion in M4 type acute myeloblastic leukemia. AB - To determine the incidence of the mixed lineage leukemia (MLL) gene rearrangements in acute myeloid leukemia (AML) without cytogenetically-detected 11q23 abnormalities, we screened 64 cases of AML at diagnosis for MLL rearrangement by FISH. Three cases (4.7%) had a MLL rearrangement detected; one was shown to have a cryptic t(11;22)(q23;q11) and another to have a t(9;11)(p21 22;q23) which had been missed by the conventional cytogenetic study. No 11q23 structural abnormality was visible in the third case. Twenty-six of the 64 cases were further studied by Southern blotting and DNA hybridization, and four of these cases (15%) were found to have MLL rearrangement: in three of these, FISH had not detected any abnormality. FISH was also used to confirm MLL involvement in eight cases of AML that had a cytogenetic abnormality at 11q23; in one of these, Southern blot did not show a rearrangement. The survival of patients with MLL abnormalities identified by cytogenetics, FISH and/or DNA analysis was significantly worse than that of patients without MLL abnormalities (event-free survival p = 0.016) although two patients with a t(9;11)(p21-22;q23) were long term survivors, consistent with this particular translocation having a better prognosis. One further case with a cytogenetic abnormality close to 11q23 was studied; it was found to have a t(10;11)(p13;q21), and the breakpoints were shown by FISH to involve the Clathrin Assembly Lymphoid Myeloid (CALM) gene at 11q21 and the AF10 gene at 10p13. Our data confirm the value of combining cytogenetic, FISH and molecular analyses to define the incidence and precise nature of MLL and 11q23 abnormalities in AML. PMID- 11911107 TI - Allergic contact dermatitis from minoxidil: study of the cross-reaction to minoxidil. PMID- 11911109 TI - Calibration monitoring of the visible and near-infrared channels of the along track scanning radiometer-2 by use of stable terrestrial sites. AB - The Along-Track Scanning Radiometer-2 (ATSR-2) is equipped with visible and near infrared channels at 1.6, 0.87, 0.66, and 0.56 microm. An in-flight visible calibration (VISCAL) system used to convert the raw signal to top-of-the atmosphere reflectances is described. To monitor the long-term stability of the VISCAL, a number of large-area stable terrestrial sites have been employed. We describe the methods used to determine the long-term drifts in the ATSR-2 onboard calibration device and evaluate the suitability of the sites for calibration monitoring. PMID- 11911110 TI - Forensic comparison of soils by bacterial community DNA profiling. AB - This preliminary investigation has shown that a soil microbial community DNA profile can be obtained from the small sample of soil recovered from the sole of a shoe, and from soil stains on clothing. We have also shown that these profiles are representative of the site of collection and therefore could potentially be used as associative evidence to prove a link between suspects and crime scenes. Soil community profiles were obtained using the T-RFLP fingerprinting method that uses fluorescent primer technology and semi-automated analysis techniques similar to those used in human DNA profiling in forensic laboratories. PMID- 11911111 TI - Intercellular adhesion molecule-1 gene polymorphisms in isolated polymyalgia rheumatica. AB - OBJECTIVE: In untreated polymyalgia rheumatica (PMR), high levels of circulating soluble intercellular adhesion molecule-1 (ICAM-1) have been observed. To investigate the clinical implication of ICAM-1 polymorphisms in isolated PMR, we examined their potential influence in an unselected series of patients. METHODS: We studied 72 patients with isolated PMR and 129 ethnically matched controls from Lugo, Spain. Patients and controls were genotyped for HLA-DRB1 and ICAM-1 polymorphism at codons 241 and 469 by molecular methods. RESULTS: The distribution of alleles and genotypes for each ICAM-1 polymorphism did not show significant differences between patients with isolated PMR and controls. There were also no associations between ICAM-1 polymorphisms and relapses of the disease. The latter was primarily associated with carriage of an HLA-DRB1*0401 allele (OR 7.2, p = 0.01), although all relapsed patients with HLA-DRB1*0401 also carried the GG genotype of the ICAM-1 polymorphism at codon 241. The presence of both HLA-DRB1*0401 and the GG241 ICAM-1 genotype gave an OR of 15.2 (p = 0.005) after correction for age and sex. CONCLUSION: Although ICAM-1 polymorphisms alone do not appear to be associated with disease severity in isolated PMR, the presence of both HLA-DRB1*0401 and the ICAM-1 codon 241 GG homozygosity was significantly associated with increased risk of relapses in these patients. PMID- 11911112 TI - Chronic fatigue syndrome and arthralgia following parvovirus B19 infection. AB - OBJECTIVE: To determine the incidence of arthralgia and fatigue complicating B19 infection, along with associated B19 markers and autoantibodies. METHODS: We studied patients with acute B19 infection (n = 51), patients followed from the time of acute B19 infection (mean 22.5 mo) (n = 39), and healthy controls (n = 50). Clinical details were collected using a questionnaire and blood was tested for B19 markers and autoantibodies. RESULTS: Acute B19 arthralgia occurred in 31 patients and was associated with female sex (p = 0.007) and age > 20 years (p = 0.02). Acute B19 fatigue occurred in 8 patients and was not significantly associated with any marker. At followup, symptoms consisted of arthralgia (n = 5), arthralgia and fatigue (n = 6), fatigue (n = 7), lymphadenopathy (n = 1), and purpura due to thrombocytopenia (n = 2). Chronic B19 arthralgia was associated with persistent B19 viremia (p = 0.029). Comparison of the B19 followup group with the controls revealed a significantly increased prevalence of arthralgia (p = 0.0002), fatigue (p < 0.0001), and all other markers. Chronic B19 arthralgia was associated with both acute B19 arthralgia (p = 0.0168) and positive ANA at acute infection (p = 0.0043). Chronic B19 fatigue was associated with acute B19 fatigue (p = 0.011). Five patients fulfilled the Centers for Disease Control criteria for a diagnosis of chronic fatigue syndrome (CFS) and one of these was negative for serum anti-B19 IgG at followup by both Western blot and immunofluorescence. However, there was no characteristic pattern of B19 markers/autoantibodies in patients with B19 associated chronic fatigue. CONCLUSION: CFS may follow acute parvovirus B19 infection; however, attribution of a case of CFS to B19 infection may be extremely difficult in the absence of serological confirmation of acute infection at fatigue onset. PMID- 11911113 TI - Study of the advancing and receding contact angles: liquid sorption as a cause of contact angle hysteresis. AB - Two types of experiments were used to study the behavior of both advancing and receding contact angles, namely the dynamic one-cycle contact angle (DOCA) and the dynamic cycling contact angle (DCCA) experiments. For the preliminary study, DOCA measurements of different liquids on different solids were performed using an automated axisymmetric drop shape analysis-profile (ADSA-P). From these experimental results, four patterns of receding contact angle were observed: (1) time-dependent receding contact angle; (2) constant receding contact angle; (3) 'stick/slip'; (4) no receding contact angle. For the purpose of illustration, results from four different solid surfaces are shown. These solids are: FC-732 coated surface; poly(methyl methacrylate/n-butyl methacrylate) [P(MMA/nBMA)]; poly(lactic acid) (DL-PLA); and poly(lactic/glycolic acid) 50/50 (DL-PLGA 50/50). Since most of the surfaces in our studies exhibit time dependence in the receding contact angle, a more extended study was conducted using only FC-732-coated surfaces to better understand the possible causes of decreasing receding contact angle and contact angle hysteresis. Contact angle measurements of 21 liquids from two homologous series (i.e. n-alkanes and 1-alcohols) and octamethylcyclotetrasiloxane (OCMTS) on FC-732-coated surfaces were performed. It is apparent that the contact angle hysteresis decreases with the chain length of the liquid. It was found that the receding contact angle equals the advancing angle when the alkane molecules are infinitely large. These results strongly suggest that the chain length and size of the liquid molecule could contribute to contact angle hysteresis phenomena. Furthermore, DCCA measurements of six liquids from the two homologous series on FC-732-coated surfaces were performed. With these experimental results, one can construe that the time dependence of contact angle hysteresis on relatively smooth and homogeneous surfaces is mainly caused by liquid retention/sorption. The results also suggested that the contact angle hysteresis will eventually approach a steady state, where the rate of liquid retention-evaporation or sorption process would balance out each other. If the existence of contact angle hysteresis can be attributed to liquid sorption/retention, one should only use the advancing contact angles (measured on a dry surface) in conjunction with Young's equation for surface energetic calculations. PMID- 11911114 TI - A Bayesian model of stereopsis depth and motion direction discrimination. AB - The extraction of stereoscopic depth from retinal disparity, and motion direction from two-frame kinematograms, requires the solution of a correspondence problem. In previous psychophysical work [Read and Eagle (2000) Vision Res 40: 3345-3358], we compared the performance of the human stereopsis and motion systems with correlated and anti-correlated stimuli. We found that, although the two systems performed similarly for narrow-band stimuli, broadband anti-correlated kinematograms produced a strong perception of reversed motion, whereas the stereograms appeared merely rivalrous. I now model these psychophysical data with a computational model of the correspondence problem based on the known properties of visual cortical cells. Noisy retinal images are filtered through a set of Fourier channels tuned to different spatial frequencies and orientations. Within each channel, a Bayesian analysis incorporating a prior preference for small disparities is used to assess the probability of each possible match. Finally, information from the different channels is combined to arrive at a judgement of stimulus disparity. Each model system--stereopsis and motion--has two free parameters: the amount of noise they are subject to, and the strength of their preference for small disparities. By adjusting these parameters independently for each system, qualitative matches are produced to psychophysical data, for both correlated and anti-correlated stimuli, across a range of spatial frequency and orientation bandwidths. The motion model is found to require much higher noise levels and a weaker preference for small disparities. This makes the motion model more tolerant of poor-quality reverse-direction false matches encountered with anti-correlated stimuli, matching the strong perception of reversed motion that humans experience with these stimuli. In contrast, the lower noise level and tighter prior preference used with the stereopsis model means that it performs close to chance with anti-correlated stimuli, in accordance with human psychophysics. Thus, the key features of the experimental data can be reproduced assuming that the motion system experiences more effective noise than the stereoscopy system and imposes a less stringent preference for small disparities. PMID- 11911115 TI - A spatial stochastic neuronal model with Ornstein-Uhlenbeck input current. AB - We consider a spatial neuron model in which the membrane potential satisfies a linear cable equation with an input current which is a dynamical random process of the Ornstein-Uhlenbeck (OU) type. This form of current may represent an approximation to that resulting from the random opening and closing of ion channels on a neuron's surface or to randomly occurring synaptic input currents with exponential decay. We compare the results for the case of an OU input with those for a purely white-noise-driven cable model. The statistical properties, including mean, variance and covariance of the voltage response to an OU process input in the absence of a threshold are determined analytically. The mean and the variance are calculated as a function of time for various synaptic input locations and for values of the ratio of the time constant of decay of the input current to the time constant of decay of the membrane voltage in the physiological range for real neurons. The limiting case of a white-noise input current is obtained as the correlation time of the OU process approaches zero. The results obtained with an OU input current can be substantially different from those in the white-noise case. Using simulation of the terms in the series representation for the solution, we estimate the interspike interval distribution for various parameter values, and determine the effects of the introduction of correlation in the synaptic input stochastic process. PMID- 11911116 TI - Improvement of vascular dysfunction and blood lipids of insulin-resistant rats by a marine oil-based phytosterol compound. AB - The syndrome that is characterized by obesity, insulin resistance, and hyperlipidemia is increasingly prevalent in all prosperous societies. It is now recognized as a major contributor to cardiovascular disease. Vascular dysfunction in the form of hypercontractility and impaired nitric oxide-mediated relaxation is a significant component of cardiovascular disease, predisposing to ischemic events. The JCR:LA-cp strain of rats exhibits all major aspects of the obesity/insulin resistance syndrome, including vascular dysfunction and ischemic lesions of the heart. Dietary lipid intake may have a marked effect on plasma lipid levels and, potentially, on vascular disease. We have investigated the effects of a novel preparation, ONC101 (a phytosterol esterified with fish oil), on plasma lipids and vascular function in the insulin-resistant JCR:LA-cp rat. Treatment of obese male rats with ONC101 from 8 to 12 wk of age resulted in no change in plasma lipid concentrations at 0.5 g/kg body weight. At the higher dose of 2.6 g/kg, plasma TG fell 50% (1.26 vs. 2.59 mmol/L, P < 0.002) and cholesterol esters were significantly reduced (1.34 vs. 1.61 mmol/L, P < 0.002). Food intake and body weights were unaffected by ONC101 treatment. At the low dose of 86 mg/kg, the hypercontractility of aortic rings in response to phenylephrine was normalized and the relaxant response to acetylcholine was significantly improved. The results indicate that ONC101 at high doses has significant hypolipidemic effects and, at very low doses, has beneficial effects on endothelial and vascular smooth muscle cell function. PMID- 11911118 TI - Lactoferrin: influences on Langerhans cells, epidermal cytokines, and cutaneous inflammation. AB - It has been suggested previously that, in addition to other biological roles, lactoferrin (LF) may display antiinflammatory properties secondary to the regulation of cytokine expression. To explore this concept further, we have here examined in human volunteers the influence of recombinant homologous LF on the migration of epidermal Langerhans cells (LC), a process that is known to be dependent upon the local availability of certain proinflammatory cytokines including tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL 1beta). In common with previous studies in mice, it was found that topical administration of LF prior to exposure at the same site to the contact sensitizer diphenylcyclopropenone resulted in a significant reduction of allergen-induced LC migration from the epidermis (measured as a function of the frequency of CD1a+ or HLA-DR+ LC found in epidermal sheets prepared from punch biopsies of the treated skin sites). However, under the same conditions of exposure, LF was unable to influence migration of LC induced by the intradermal administration of TNF-alpha data consistent with the hypothesis that one action of LF in the skin is to regulate the local production of this cytokine. Further support for this hypothesis was derived from experiments conducted with IL-1beta. This cytokine is also able to induce the mobilization of LC following intradermal injection, although in this case, migration is known to be dependent upon the de novo production of TNF-alpha. We observed that prior exposure to LF resulted in a substantial inhibition of IL-1beta-induced LC migration, data again consistent with the regulation of TNF-alpha production by LF. Collectively, these results support the view that LF is able to influence cutaneous immune and inflammatory processes secondary to regulation of the production of TNF-alpha and possibly other cytokines. PMID- 11911117 TI - Locally and systemically active glucocorticosteroids modify intestinal absorption of lipids in rats. AB - Orally administered systemically active steroids enhance the digestive and absorptive functions of the intestine, but their effect on lipid uptake is unknown. The effect of the locally acting steroid budesonide on intestinal absorptive function also is unknown. Accordingly, this study was undertaken to assess the influence of 4 wk of treatment of weaning male rats with a daily oral gavage of budesonide (BUD), prednisone (PRED), or control vehicle on the jejunal and ileal uptake of fatty acids and cholesterol. BUD enhanced jejunal uptake of oleic acid and ileal uptake of linoleic acid. PRED increased jejunal uptake of cholesterol and ileal uptake of lauric, palmitic, linoleic, and linolenic acids. Higher doses of BUD (up to 1 mg/kg) given to adult rats for 2 wk further increased the uptake of some lipids. The changes in the uptake of lipids were not due to variations in the weight of the intestinal mucosa or in the animals' food intake. Ileal ornithine decarboxylase mRNA expression was increased with PRED, but there were no steroid-associated changes in the expression of the mRNA of the early response genes c-myc, c-jun, or c-fos or of proglucagon, the liver fatty acid-binding protein (FABP), the ileal lipid-binding protein, tumor necrosis factor alpha, interleukin 2 (IL-2), IL-6, or IL-10. In summary, treatment of weanling rats with BUD and PRED enhances the uptake of some lipids by a process that is independent of the effects of early response genes and genes encoding cytokines, proglucagon, and FABP. PMID- 11911119 TI - Genome size and microsatellites: the effect of nuclear size on amplification potential. AB - Although the frequency of microsatellite DNA regions generally increases with increasing genome size, genome size has a negative effect on polymerase chain reaction (PCR) amplification. Thus, researchers developing sets of PCR primers, as is commonly done for microsatellite DNA regions, may encounter greater difficulty when working with species that have larger genomes. I investigated the effect of genome size on overall amplification success using data from nine different metazoan taxa. The proportion of primer sets that did not amplify PCR products was strongly and positively correlated with the haploid C value of the target species. Increasing genome size may affect amplification success negatively because of a decrease in target:nontarget DNA or by dilution of the available primer pool by nonspecific binding. PMID- 11911120 TI - Molecular evolution of ribosomal intergenic spacers in Odontophrynus americanus 2n and 4n (Amphibia: Anura). AB - Ribosomal intergenic spacers (IGSs) of Odontophrynus americanus 2n and 4n were cloned, restriction mapped, and partially sequenced. Three distinct regions, namely alpha, beta, and delta, were identified in the IGSs. The alpha and beta regions flanked the 28S and 18S rRNA genes, respectively, conserving an identical restriction pattern at each ploidy level. The delta region, located between alpha and beta, was highly variable in size and restriction pattern, enclosing different BamHI subrepeats (B-SR), 87- to 530-bp-long. Sequence analysis showed that B-SRs were composed mainly of different arrangements of similar blocks of sequences. Another family of repetitive sequences was found in the delta region, clustered inside large BamHI fragments. These subrepeats are 189-bp-long and, although very similar in diploid and tetraploid IGSs, show a pattern of concerted evolution. A hypothetical functional role for the 189-bp repeats is discussed in view of their predicted secondary structure and presence of potential E2 binding sites inside diploid subrepeats. Although the same structural elements were present both in diploid and tetraploid IGSs, the higher level of repeatability of tetraploid IGSs suggests that common ancestor sequences have undergone several rounds of amplification after O. americanus polyploidy. PMID- 11911121 TI - Effect of personality dimensions on the diurnal pattern of motor activity. AB - The effects of personality dimensions, age, and gender on 24-hour motor activity patterns were studied in 101 healthy subjects between 20-70 years. We measured motor activity by wrist-actigraphy and personality dimensions by the Tridimensional Personality Questionnaire (TPQ) of Cloninger. Random Regression Models were used to estimate the effects of personality dimensions, age, and gender on the diurnal pattern of motor activity. Harm Avoidance as well as Reward Dependence influenced the overall level of motor activity, independent of age and gender. Subjects high on Harm Avoidance showed lower activity levels than subjects low on Harm Avoidance, whereas subjects high on Reward Dependence had higher overall levels of motor activity than subjects low on Reward Dependence. Older subjects were found to be less active than younger subjects, but the activity level did not differ between males and females. PMID- 11911123 TI - Compensation monitor. Executive bonuses: health care takes care of its own. PMID- 11911122 TI - Unexplained deaths following knee surgery--Minnesota, November 2001. PMID- 11911124 TI - Molecular classification of psoriasis disease-associated genes through pharmacogenomic expression profiling. AB - Psoriasis is recognized as the most common T cell-mediated inflammatory disease in humans. Genetic linkage to as many as six distinct disease loci has been established but the molecular etiology and genetics remain unknown. To begin to identify psoriasis disease-related genes and construct in vivo pathways of the inflammatory process, a genome-wide expression screen of multiple psoriasis patients was undertaken. A comprehensive list of 159 genes that define psoriasis in molecular terms was generated; numerous genes in this set mapped to six different disease-associated loci. To further interpret the functional role of this gene set in the disease process, a longitudinal pharmacogenomic study was initiated to understand how expression levels of these transcripts are altered following patient treatment with therapeutic agents that antagonize calcineurin or NF-KB pathways. Transcript levels for a subset of these 159 genes changed significantly in those patients who responded to therapy and many of the changes preceded clinical improvement. The disease-related gene map provides new insights into the pathogenesis of psoriasis, wound healing and cellular-immune reactions occurring in human skin as well as other T cell-mediated autoimmune diseases. In addition, it provides a set of candidate genes that may serve as novel therapeutic intervention points as well as surrogate and predictive markers of treatment outcome. PMID- 11911125 TI - Good standards should not need legislation. PMID- 11911126 TI - Nursing specialties still influenced by tradition. PMID- 11911127 TI - Care of elderly people and nursing standards. PMID- 11911128 TI - Ageist attitudes also rife among nurses. PMID- 11911129 TI - From the Centers for Disease Control and Prevention. Rashes among schoolchildren- 14 states, October 4, 2001-February 27, 2002. PMID- 11911130 TI - From the Centers for Disease Control and Prevention. Recent trends in mortality rates for four major cancers, by sex and race/ethnicity--United States, 1990 1998. PMID- 11911131 TI - JAMA patient page. Celiac disease. PMID- 11911132 TI - An interview with Professor Thomas Tursz. PMID- 11911133 TI - Prevention of venous thromboembolism with fondaparinux. PMID- 11911134 TI - Prevention of venous thromboembolism with fondaparinux. PMID- 11911135 TI - Prevention of venous thromboembolism with fondaparinux. PMID- 11911136 TI - Recognition and management of anthrax. PMID- 11911137 TI - Recognition and management of anthrax. PMID- 11911138 TI - Recognition and management of anthrax. PMID- 11911139 TI - Sudden death due to cardiac arrhythmias. PMID- 11911140 TI - Haemodynamic changes caused in rats by prolonged accelerations. AB - Experiments were performed on 60 rats undergoing about 5 Gz acceleration. The animals were killed in liquid nitrogen at various times after centrifugation (time of centrifugation 1 hour). Before centrifugation 131I-labelled albumin was injected intravenously and after the experiment radioactivity of muscles from the extremities was measured. The following conclusions were drawn from the experiments: (i) About 5 Gz acceleration in the longitudinal axis results in blood displacement to the hind limbs with extravascular leakage of albumin. (ii) Immediately after centrifugation lowering of blood amount in the hind limbs occurred. During the first 15 minutes this decrease progresses very quickly, while later it is much smaller. Nevertheless even 3 hours after centrifugation there was only partial return to normality. (iii) Transudation of albumin in the hind limbs lasts a few hours after centrifugation. (iv) The haemodynamic changes caused by centrifugation consist of blood stasis and extravascular leakage of proteins. PMID- 11911141 TI - Effect of hypergravity and hyperthermia on antidiuretic hormone secretion. AB - The effect of acceleration and hyperthermia on the antidiuretic hormone secretion (ADH) was investigated in rats both separately and simultaneously. The two conditions of stress elicited a rise in plasma ADH concentration in proportion to their intensity. Concomitant exposure to the two factors produced an additional effect. The parallel histochemical studies using methods for demonstrating RNA, proteins and the neurosecretory material in the supraoptic nucleus, showed the synthesis and depletion of the hormone content in correlation with the plasma concentration of ADH. PMID- 11911142 TI - Metabolic responses of monkeys to increased gravitational fields. AB - A depletion of body fat has been reported by several investigators for various species of homoiotherms exposed to chronic acceleration. Generally the effect is proportional to field strength, and over the size range examined (0.25-5 kg) it becomes more severe in larger animals. There is some evidence that this may be the result of increased secretion of a pituitary hormone, the fat mobilizing substance (FMS). An exception to the defatting effect of chronic acceleration was observed in mature pig-tailed monkeys (Macaca nemestrina)--although they exhibited other changes generally found in homoiotherms exposed to hyperdynamic environments. It is considered that this unusual primate response to chronic acceleration may result from a different geometry of the central nervous system, and its effect upon gravity load distribution in the lower part of the brain. PMID- 11911143 TI - Gravitational stress and exercise. AB - This paper deals with the influence of G forces on certain circulatory and metabolic adjustments to dynamic exercise. Leg exercise in the sitting position increases systemic mean pressure both at normal and at increased G, thus improving G tolerance. When the force of gravity is increased to three times its normal value, leg exercise produces a much more marked increment in stroke volume than in heart rate, which contrasts with the case at normal gravity. Beta adrenergic blockade, which strongly inhibits the heart rate response to gravitational stress, but leaves the systemic mean pressure and G tolerance unaffected, exaggerates the dominance of the stroke volume increase in exercise at increased G. With leg exercise at increasing work loads at 3G, the O2 uptake tends to level off and the arterial lactates to rise at a lower load than at normal gravity, indicating that the upper limit for the O2 uptake is lowered by the G force. A progressive increase of the venous admixture in the lungs suggests that the primary limitation of the O2 transport occurs in the pulmonary gas exchange. PMID- 11911145 TI - Histological studies on the vestibular organ of frog embryos and larvae after simulated weightlessness. AB - The ontology of gravity receptors is discussed. As well as phylogenetic aspects of the question, we may ask whether the different gravity receptors in plants and animals derive from the same source; did they evolve from a general sensitivity of the living cell or do they represent parallel inventions. One may also ask whether during ontogeny of the receptor it receives information from gravity or whether the development of function of the organ is an autonomous process. In many cases ontogeny shows traces of phylogeny. Statoliths, e.g. the differentiating statolith membranes of the frog embryo, look somewhat like sedimentational structures. Furthermore the amphibian embryo in its relatively early stages takes up a special position in relation to the direction of gravity. Which mechanism is involved here? The materials studied were frog embryos and larvae which developed within a fast rotating clinostat. The results may help to answer the question as to whether the differentiation of the frog statolith membrane is an autonomous process. PMID- 11911144 TI - The role of gravity in the phylogeny of structure and function in animal sensors of spatial orientation, and their predicted action in weightlessness. AB - The evolution of the structural, functional and cytochemical organization of the gravity receptor which determines a body position in the gravitational field of the earth by means of muscular regulation was traced both invertebrates and vertebrates, using electron microscopic and histochemical methods. In the course of evolution of vertebrates, the specialized gravity receptor-statocyst which, as a rule, consists of primary sensory cells and supplies otoliths, is formed. In vertebrates, there exists a vestibular apparatus made up of secondary sensory cells and also having otoliths. The receptor cells, both of statocysts and the vestibular apparatus, are supplied with special antennas (kinocilia and stereocilia). Deviation of the antennas stimulated by displacement of the otoliths resulting from locomotor activity of animals leads to excitation of the receptor cells. When exposed to a modified gravitational field (linear accelerations of 10 g, for 3 min), the receptor cells of the vestibular apparatus, in all classes of vertebrates, show progressive changes in RNA content and protein synthesis (increase followed by decrease) which return to normal only after 12 days. Thus, immediate transfer of animals and man from acceleration to weightlessness appears to be a reason for movement disease. The above consideration showed the need for an experiment in which an animal (with its vestibular apparatus) which had not undergone previous accelerations, would be exposed to weightlessness. Frog embryos, Rana temporaria, at the stage preceding the organogenesis, when the vestibular apparatus and other organs were lacking, were chosen as a suitable subject. Frog embryos at the stage of an early gastrula were placed in a special container Emkon aboard the Soyuz 10 spacecraft. After short accelerations, they were exposed to weightlessness for 44 hours. The embryos were allowed to continue to develop to the stage of early tail bud. The experimental embryos showed normally developed acoustic vesicles and vestibular ganglia. Clear differentiation of the receptor cells with antennas (kinocilia and stereocilia) was found in the acoustic vesicles. Thus, in weightlessness, vestibular apparatus develops just as well as in the gravitational field of the earth. However, only a much longer stay in weightlessness conditions will indicate whether there are any changes in the structural, cytochemical and functional organization of vestibular apparatus. The similarity in the structural, functional and cytochemical organization of the gravity receptor in vertebrates and invertebrates appears to allow the prediction of the behaviour of the gravity receptor as a whole, and of its receptor elements, both in normal and changed gravitational fields. The first attempts were carried out only on the vestibular apparatus of vertebrates. PMID- 11911146 TI - Viability of Bacillus subtilis spores exposed to space environment in the M-191 experiment system aboard Apollo 16. AB - During the Apollo 16 space flight, in the experiment system M-191, (microbial response to space environment) spores of Bacillus subtilis 168 were exposed to space vacuum or solar UV irradiation with a peak wavelength of 254 nm or both. The effects of these space factors on the colony-forming ability of the spores were studied. It was found (i) that space vacuum alone did not affect the survival of pre-dried spores; (ii) that space vacuum in combination with solar UV irradiation with a peak wavelength of 254 nm had a synergistic effect, which may by attributed to a UV supersensitivity of the spores during vacuum exposure. These results agreed with findings of simulation experiments on earth. It was concluded that air dried spores may survive exposure to space vacuum if shielded against solar UV irradiation. PMID- 11911147 TI - Preliminary results on the action of cosmic heavy ions on the development of eggs of Artemia salina. AB - The Biostack experiment (Principal Investigators: H. Bucker) aboard Apollo 16 contained one unit with eggs of the brine shrimp Artemia salina. The eggs were fixed in polyvinyl alcohol foils. Nuclear emulsions and plastics were used as track detectors. The development of 260 eggs hit by cosmic heavy ions was investigated. It differed significantly from the development of the flight controls--eggs flown in the Biostack but not hit by heavy ions--and of the ground controls. From these results it was concluded that a hit of a single heavy ion may injure the encysted blastula. This damage was found to influence the gastrula formation and even the hatching process of the nauplius. Abnormalities of the orthonauplius were observed during the development of the hit eggs. These are shortened extremities, or abnormal thorax or abdomen. In addition, in the Biostack II aboard Apollo 17 eggs of Tribolium confusum and Carausius morosus were included. The influence of single heavy ions on the development process of these highly organized insects was investigated. PMID- 11911148 TI - Microbial studies in the Biostack experiment of the Apollo 16 mission: germination and outgrowth of single Bacillus subtilis spores hit by cosmic HZE particles. AB - Bacillus subtilis spores were flown in the Biostack experiment aboard the Apollo 16 command module. The spores embedded in plastic foils were stacked between physical track detectors. The energy loss spectrum of the heavy particles of cosmic radiation was determined. Biological studies were restricted to the high energy loss component of these particles. Spores that had received single hits whose positions were determined with a typical accuracy of +/- 1 micrometers, were investigated for radiation effects on germination and outgrowth. It was found that germination was not influenced by a hit by an HZE particle, but outgrowth was reduced significantly. PMID- 11911149 TI - Study of cosmic ray effects on Artemia salina eggs during the Apollo 16 and 17 flights. AB - We have used Artemia salina eggs, embedded in polyvinyl alcohol, to study the biological effects of heavy ions of cosmic rays. Each biological layer was sandwiched between track detectors. Hit eggs by heavy ions show a great inhibition of their developmental ability. A lower inhibition is observed for eggs that were flown but not hit. Simulation experiments are in progress to determine the factors responsible for inhibition of eggs that were not hit and to improve our knowledge of cellular damage induced by heavy ions. PMID- 11911150 TI - European surveillance shows north-south divide in resistant bacteria. PMID- 11911151 TI - EU to tighten rules on vitamin pills. PMID- 11911152 TI - Psychologists allowed to prescribe drugs for mental illness. PMID- 11911153 TI - US doctors launch campaign on malpractice claims. PMID- 11911154 TI - MMR vaccine debate. General practitioners' two roles are not in conflict with MMR immunisation. PMID- 11911155 TI - MMR vaccine debate. Single measles vaccine should be allowed. PMID- 11911156 TI - MMR vaccine debate. There is no room for lingering doubt. PMID- 11911157 TI - MMR vaccine debate. Competing interests need to be declared. PMID- 11911158 TI - Physician assistants. American idea of physician assistants can be anglicised. PMID- 11911159 TI - European working time directive for doctors in training. Profession needs to modernise surgical training. PMID- 11911160 TI - Heisenberg and Bohr--another view. PMID- 11911161 TI - Survey of foreign recipients of U.S. Ph.D.'s. PMID- 11911162 TI - Letter of appreciation. PMID- 11911163 TI - Cholesterol's role in synapse formation. PMID- 11911164 TI - Detection of biogenic CO production above vascular cell cultures using a near room-temperature QC-DFB laser. AB - We report the first application of pulsed, near-room-temperature quantum cascade laser technology to the continuous detection of biogenic CO production rates above viable cultures of vascular smooth muscle cells. A computer-controlled sequence of measurements over a 9-h period was obtained, resulting in a minimum detectable CO production of 20 ppb in a 1-m optical path above a standard cell culture flask. Data-processing procedures for real-time monitoring of both biogenic and ambient atmospheric CO concentrations are described. PMID- 11911165 TI - Treatment of malignant esophagorespiratory fistulas with covered stents. PMID- 11911166 TI - Gastrointestinal fistulas: treatment with covered stents. PMID- 11911167 TI - Cervical esophageal webs: association with gastroesophageal reflux. AB - BACKGROUND: We investigated whether there is a significant association between cervical esophageal webs and gastroesophageal reflux on pharyngoesophagography. METHODS: We studied 50 patients with cervical esophageal webs on pharyngoesophagrams and 50 control subjects. The control group was matched to the webs group for age, sex, and symptomatology. Patients with cervical esophageal webs and controls were compared to determine the prevalence of gastroesophageal reflux, hiatal hernias, reflux esophagitis, and abnormal esophageal motility. Pearson's chi-square test was used to determine any statistically significant differences in the frequencies of these findings between groups. RESULTS: Thirty nine (78%) of 50 patients with cervical esophageal webs versus 27 (54%) of 50 patients in the control group had gastroesophageal reflux (p = 0.01). When patients were classified based on degree of gastroesophageal reflux, 22 (44%) of 50 patients with cervical esophageal webs versus 21 (42%) of 50 controls had mild reflux (p = 0.84), whereas 17 (34%) of 50 patients with webs versus six (12%) of 50 controls (p < 0.009) had moderate/marked reflux. Thus, the prevalence of moderate/marked gastroesophageal reflux was significantly greater in patients with webs than in the controls. However, no significant differences were found in the prevalence of mild gastroesophageal reflux, hiatal hernias, reflux esophagitis, or abnormal esophageal motility. CONCLUSION: We found a significant association between cervical esophageal webs and gastroesophageal reflux independent of age, sex, or symptomatology. Radiologists should be aware of this association, so that patients with cervical esophageal webs on pharyngoesophagography are evaluated for gastroesophageal reflux at the time of the barium study or advised to undergo further testing for gastroesophageal reflux disease. PMID- 11911168 TI - The need for hypoglycemia detection and prevention in type 1 diabetes. PMID- 11911169 TI - The impact of initiatives in education, self-management training, and computer assisted self-care on outcomes in diabetes disease management. AB - The purpose of this work is to elucidate the advantages and disadvantages arising from three distinct diabetes disease management initiatives in a managed care setting. The initiatives included (1) education alone, (2) education with self management training, and (3) education with computer-assisted self-care. Outcomes of interest were changes in glycated hemoglobin (HbA1c), body weight, and costs of care in each cohort of care recipients. A total of 978 health plan members with diabetes within a mixed model HMO were included in the initiatives for improving blood glucose control. HbA1c was measured at baseline and at 3 and 12 months, body weight at baseline and 12 months, and costs of care over 1 year. Costs were derived from suppliers and the health plan. The design is a longitudinal observation study. With the edu-cation-alone initiative, costs, HbA1c, and body weight were unchanged. When education is supplemented with ongoing self-management training, HbA1c fell 1.1% (p < 0.01), body weight rose by 11 kg (p < 0.01), and costs for care increased by $18 per member per month. When education is supplemented with ongoing computer-assisted self-care, HbA1c also dropped by 1.1% (p < 0.01), body weight was unchanged (p > 0.4), and costs for care were $1.31 per member per month. All initiatives improved glycated hemoglobin. Other outcomes must therefore be considered. Among the initiatives, this study elucidated significant differences in body weight and costs. Therefore, in choosing a diabetes disease management program, it would appear that costs should be the primary consideration and methodologies that control body weight should be a priority. PMID- 11911170 TI - What we can really expect from telemedicine in intensive diabetes treatment: results from 3-year study on type 1 pregnant diabetic women. AB - Existing standards of the management of the diabetic patients are not efficient enough, and further improvement is needed. The major objective of this paper is to present and discuss the therapeutic effectiveness of an intensive care telematic system designed and applied for intensive treatment of pregnant type 1 diabetic women. The developed system operates automatically, every night transferring all the data recorded during the day in the patient's glucometer memory to a central clinical unit. In order to assess the efficiency of the designed and developed system, a 3-year randomized prospective clinical trial was conducted, using the study group and the control group, each consisting of 15 pregnant type 1 diabetic women. All patients were treated by the same diabetologist. In the presented analysis, two indices calculated weekly were used for the assessment of glycemic control: MBG represents mean blood glucose level, and the universal J-index is sensitive to the glycemic level and glycemic variations. The most important results from the study concern: (a) better glycemic control in the study group in comparison with the control group during the course of treatment, as assessed by the average differences of the MBG and J indices calculated weekly (n = 24) (deltaMBG = -3.2 +/- 4.3 mg/dL, p = 0.0016, deltaJ = -1.4 +/- 2.3, p = 0.0065); (b) much more similar results in glycemic control among members of the study group compared to each other, than among members of the control group compared to each other, as indicated by significantly lower variations of the applied glycemic control indices (SDMBG: 11.9 vs. 18.7 mg/dL, p = 0.0498; SDJ: 6.5 vs. 10.9, p = 0.0318); (c) the observed tendency of a better glycemic control for patients with a lower level of intelligence (IQ < 100) supported by the telematic system in comparison with all other assessed groups of patients. The last result was not statistically significant (p > 0.05). This telematic intensive care system improved the effectiveness of diabetes treatment during pregnancy. It also allows the diabetologist's strategy to be much more precise than if it were conducted without telematic support. This telematic system is inexpensive and simple in use. PMID- 11911172 TI - Computer-based remote diabetes education for school personnel. AB - Teachers are expected to respond quickly and accurately to any diabetes incident that may occur to children in the school setting. Access to diabetes information is crucial for student safety, health, academic achievement, and social competence. This paper describes a technique to provide Web-based diabetes information using computer audio and video to enrich a text-based training experience. Two groups of teachers were presented with diabetes training material via either paper or a Web-based computer system. Both groups were then evaluated for diabetes knowledge and satisfaction. Subjects using the Web-based system had significantly (t = 2.22; p < 0.033) higher knowledge scores (72.5% versus 66.4% correct) and were significantly (t = 3.9; p < 0.001) more satisfied with the training session (4.2 versus 3.1 on a five-point scale) than subjects who used paper documents traditionally used for teacher training. With the advantages in learning and the reduced cost of a Web-based system, diabetes distance education is a viable and desirable alternative to paper-based diabetes education. PMID- 11911171 TI - Accuracy of the hemocue portable glucose analyzer in a large nonhomogeneous population. AB - Several studies have reported inconsistent results between HemoCue (HC) whole blood glucose measurements compared to plasma glucose. We selected a large patient population with diverse pathologies and healthy volunteers to evaluate HC. For this comparison, whole blood glucose concentration was measured using HC and referenced to laboratory plasma glucose. The population (n = 512) included healthy volunteers, diabetics, and patients with heart failure, liver failure, renal failure, renal and liver transplant, and other chronic diseases. Patients were on a wide variety of medications, vitamins, and food supplements. Venous blood samples were collected in tubes containing potassium oxalate and sodium fluoride. Comparison of the results was made using the method of Bland and Altman and ANOVA at three selected glucose ranges. The glucose measurement ([HC + laboratory]/2) ranges were 24-75, 76-129, and 130-404 mg/dL. A positive bias for all three glucose ranges was observed: 38 +/- 17 mg/dL for the high glucose group compared to 24 +/- 9 mg/dL and 22 +/- 10 mg/dL for the middle and low groups, respectively. In the high glucose group 90% of the values were within 10% (R = 0.97) of the laboratory reference values compared to 81% and 55% in the normal and low glucose groups, respectively. HC glucose measurements were generally within two SD from the laboratory plasma reference. HC consistently yielded lower whole blood glucose measurements than plasma with the largest differences seen in the low glucose range (29%). HC measured more consistently at the higher glucose concentrations and was 16% lower than plasma, although the mean absolute error was highest for that range. No significant effects in the bias could be attributed to disease while possible effects from instrument modifications by the manufacturer remain uncertain. PMID- 11911173 TI - DYN 12, a small molecule inhibitor of the enzyme amadorase, lowers plasma 3 deoxyglucosone levels in diabetic rats. AB - 3-Deoxyglucosone (3DG) is a highly reactive alpha-dicarbonyl sugar and potent protein cross-linker that is important in the formation of advanced glycation end products (AGEs), which have been postulated to lead to the development of diabetic complications. (1) Reducing 3DG levels in diabetics is a potentially effective therapy to slow the development of diabetic complications. Standard biochemical methods were used to isolate, identify, and characterize the enzyme responsible for the production of 3DG, in order to develop an effective therapeutic agent against this target. We have purified and characterized Amadorase, a fructosamine-3-kinase, and demonstrated both in vitro and in vivo that it is responsible for the production of 3-deoxyglucosone (3DG). A small molecule inhibitor of Amadorase, DYN 12, significantly lowered plasma levels of 3DG in diabetic (by 46%, p = 0.0116) and normal (by 43%, p = 0.0024) rats. These data are the first indications that it is possible to significantly reduce 3DG production in diabetics and thus possibly reduce the development of diabetic complications. PMID- 11911174 TI - Imaging in vivo brain-hormone interaction in the control of eating and obesity. AB - The field of neuroimaging has made great progress in the mapping of human brain function. In this article, we present a functional magnetic resonance imaging (fMRI) study on the hypothalamic regulation of satiety and its relationship with obesity. The fMRI techniques have been proven invaluable for analyzing changes in brain activity that are associated with most sensorimotor and cognitive functions. However, few studies have been successful in the delineation of the interaction between the central nervous system and the endocrine system, due to the lack of suitable mapping methods which can pinpoint the onset of changes in neuronal activity (e.g., those following eating or drug intake). We have recently introduced a new fMRI method known as temporal clustering analysis (TCA) for dynamically tracking the time course of brain activation. Along with simultaneous blood sampling for the circulating hormone levels, the fMRI techniques with TCA may provide an integrated view of the nervous and endocrine systems in vivo, and thus greatly enhance our understanding of the complex interplay between neural events and hormonal signals. PMID- 11911175 TI - Dose accuracy testing of the humalog/ humulin insulin pen device. AB - The primary purpose of the study was to determine whether pen users would challenge the insufficient remaining dose (IRD) stop mechanism with sufficient force to affect the dose accuracy of the final dose. The secondary purpose was to determine the participant's positive and negative impressions of the Humalog/Humulin pen and the likelihood of using the new prefilled pen. Three different modifications to the prefilled pen's IRD stop feature were made. These three pen models then underwent environmental dose accuracy testing at various temperatures and humidities, and user dose accuracy testing by 64 patients with diabetes. Evaluation also involved challenging the IRD stop at various dialing torques. Thirty pens from each model were tested to failure of the IRD stop. A model of the prefilled pen was selected for commercialization that met the dose accuracy targets of +/- 1 unit (U) for insulin doses less than 20 U and +/- 5% of dose volume for doses equal to or over 20 U. The selected pen model was superior at the minimum (1 unit), median (30 unit) and maximum (60 unit) dose volumes. Also 92% (n = 59) of patients interviewed felt that the stop mechanism for the final dose was clear. Extensive testing in the development of a prefilled insulin delivery device demonstrates an accurate and reliable medical device. PMID- 11911176 TI - The pen is mightier than the needle (and syringe). PMID- 11911177 TI - New therapies for the pregnant patient with diabetes. AB - Gestational diabetes complicates 3-5% of pregnancies. Of diabetes seen during pregnancy, 10% is pregestational and the remaining 90% represents gestational diabetes. (1,2) Pregnancy in women with pregestational diabetes is especially high risk. Spontaneous abortion, preterm labor, congenital malformations, preeclampsia, macrosomia, birth injury, and cesarean section are all increased in these pregnancies. Deterioration of maternal health during pregnancy, especially in the setting of diabetes-induced end-organ disease, is a real concern. Vigilant surveillance and management of associated disorders such as retinopathy, nephropathy, and chronic hypertension are required. During the preinsulin era, maternal and perinatal mortality in pregnancies complicated by pregestational diabetes was approximately 50%. (1,2) Although modern obstetrical management and the appropriate use of insulin have dramatically improved maternal-fetal outcomes, pregnant patient with diabetes remains at increased risk for complications. There is no doubt that optimizing maternal glucose control is a key element in avoiding established perinatal risks. The most effective means to accomplish this control are topics of active research. Further, hormonal changes during pregnancy can make glycemic control difficult even for the most compliant and educated patient. This paper discusses several new approaches, either currently in practice or under consideration, to pregnancies complicated by diabetes, including oral hypoglycemic agents, lispro, the insulin pump, and transplantation. PMID- 11911178 TI - Using genetic advances to investigate diabetes in Tasmania. PMID- 11911180 TI - The future of sensor technologies in diabetes: impact on healthcare delivery. PMID- 11911179 TI - The impact of continuous glucose monitoring on hospital point-of-care testing programs. PMID- 11911182 TI - Diabetes technology news. PMID- 11911181 TI - Use of the AIDA diabetes simulation software--www.2aida.org--as an interactive educational tool for teaching student nurses. AB - In previous "Diabetes Information Technology & WebWatch" columns, various user experiences with an interactive educational virtual diabetes patient simulator, called AIDA, have been documented. The simulator is available free of charge from www.2aida.org on the Web. In the 5+ years since the program was first made available on the Internet, over 125,000 people have visited the AIDA Website and over 27,000 copies of the program have been downloaded, gratis. User comments that have been received about the program have highlighted some of the many and varied ways in which a range of people have been applying the diabetes simulations in their own particular situations and practices. Inevitably, up to now, a great deal of attention has focused on use of the program by individuals with diabetes and their relatives, as well as by health-care professionals such as diabetologists/endocrinologists, diabetes educators, and primary care physicians (general practitioners [GPs]). However, an important group of health carers involved in the provision of day-to-day care for many people with diabetes are nurses. The current "Diabetes Information Technology & WebWatch" column overviews a workshop held in June 2001 in Italy to gain experience with application of the AIDA diabetes simulation approach as a teaching tool for student nurses. Feedback obtained from participants attending the workshop was generally very positive, with the student nurses reporting the simulation approach to be both of interest and of use. Further workshops involving other health-care students and professionals-in particular, medical students and qualified nurses-are planned. PMID- 11911183 TI - FimZ binds the Salmonella typhimurium fimA promoter region and may regulate its own expression with FimY. AB - The FimZ protein, an activator of FimA production in Salmonella typhimurium, acts in conjunction with FimY to facilitate the expression of type 1 fimbriae. The predicted amino acid sequence of FimZ suggests that this protein may be a DNA binding protein related to BvgA, a sensory regulator of virulence gene expression in Bordetella pertussis. Purification of FimZ following overexpression of the protein by a strong inducible promoter and gel mobility shift assays confirm that FimZ is a 25-kDa polypeptide that binds to the promoter region offimA. The region of DNA protected from DNase I digestion by FimZ binding is located between 47 and 98 nucleotides upstream from thefimA transcription initiation site. This region possesses a pair of 7-base pair tandem repeats, of which at least one is necessary for FimZ binding. One copy of the 7-base pair sequence is also located in thefimZ promoter region. In addition, expression from afimZ-lacZ reporter construct confirms that FimZ plays a role in its own expression. Both FimZ and FimY are required for high-level expression of FimZ, which suggests that these two fimbrial proteins are involved in regulating both FimA and FimZ. PMID- 11911184 TI - Two-dimensional analysis of exoproteins of methicillin-resistant Staphylococcus aureus (MRSA) for possible epidemiological applications. AB - We applied two-dimensional gel electrophoresis (2-DE) to the total exoproteins secreted from pathogenic MRSA strains and identified major protein spots by N terminal amino acid sequence analysis. In approximately 300 to 500 spots visualized on each gel, various exoproteins and cell-associated proteins were identified and their sites on the gels confirmed for construction of a reference map. Major exotoxins such as enterotoxins SEA, SEB, and SEC,, toxic shock syndrome toxin-1 (TSST-1), and hemolysins were distributed in the region of pI 6.8 to 8.1 and MW 21 to 35 kDa. Although the differences between calculated and observed values of pI and MW were relatively small in each exoprotein, those of several proteins including alpha-hemolysin and SEB were considerably deviated from the positions of the expected values. Some exoproteins were detected as multiple spots. These included beta-hemolysin, enterotoxins SEA, SEB, and SEC3, glutamic acid-specific endopeptidase, glycerophosphoryl diester phosphodiesterase and triacylglycerol lipase. The multiple spots of these exoproteins may be generated by the action of own proteases. Certain similarities of 2-DE patterns among strains belonging to the same coagulase types were observed. On the basis of 2-DE image analysis, coagulase type II strains secreted somewhat larger amounts of SEB and SEC3 as well as TSST-1 than the strains belonging to other coagulase types. Taken together, 2-DE analysis of exoproteins is applicable to epidemiological studies for MRSA, as compared with pulsed field gel electrophoresis of restricted chromosomal DNA. PMID- 11911186 TI - Promoted cell death of cells expressing human MxA by influenza virus infection. AB - Interferon-inducible MxA protein plays a crucial role in cellular protection from RNA virus infection, although the protection mechanism is not completely clarified. Here, we examined effects of MxA on either uninfected or influenza virus A/PR/8/34-infected cells. Viral protein synthesis was reduced in cells expressing MxA. Under serum-starved conditions, not only viral but also cellular protein synthesis was reduced by expression of MxA. Of interest is that MxA promoted cell death induced by apoptotic stimuli as well as influenza virus infection. These results lead to a possibility that MxA suppresses multiplication of influenza virus by affecting cellular functions including the apoptotic pathway. PMID- 11911185 TI - Immunogenicity of a recombinant MVA and a DNA vaccine for Japanese encephalitis virus in swine. AB - We previously reported that mice immunized with recombinant modified vaccinia virus Ankara (MVA) encoding Japanese encephalitis virus (JEV) prM and E genes were completely protected against JEV challenge (Nam, J.H., Wyatt, L.S., Chae, S.L., Cho, H.W., Park, Y.K., Moss, B. Vaccine 1999,17: 261-268). In this study, we examined the immunogenicity in swine of this recombinant MVA (vJH9) or a DNA vaccine (pcJH-1) expressing the same JEV genes. Although the booster effect in mice with a combination of vJH9, pcJH-1 and inactivated JEV commercial vaccine was not apparent by measuring JEV antibodies, the recombinant MVA vaccine (vJH9) and the DNA vaccine (pcJH-l) efficiently produced neutralizing antibodies in swine and 2 doses of each showed a booster effect in mice and swine. Therefore, both vJH9 and pcJH-1 are good candidates for a second generation JEV vaccine. PMID- 11911187 TI - Chemokine production by rat macrophages stimulated with streptolysin O from Streptococcus pyogenes. AB - The contribution of streptolysin O (SLO) from Streptococcus pyogenes to neutrophil infiltration in inflammatory lesions was determined by production of cytokine-induced neutrophil chemoattractant (CINC)-1, -2 and -3, and macrophage inflammatory protein (MIP)-1alpha by rat macrophages stimulated with SLO in culture. Active SLO induced the production of CINCs and MIP-1alpha in dose- and time-dependent manners. These inductions were ascertained by chemokine mRNA expression in macrophages. Streptolysin S was without effect. The SLO-cholesterol complex induced the chemokine production in proportion to the residual hemolytic activity of the complex. In addition, the effects of SLO on the chemokine production were confirmed by the injection of active SLO into the preformed air pouch on the back of rats. The infiltration of neutrophils into the pouch fluid (exudate) increased steadily with a lag phase of about 2 hr. The major chemokine found in exudates was MIP-1alpha but not CINCs. In this study, it became clear that active SLO, but not the inactive one, contributed to the production of MIP 1alpha and CINCs in the conditioned medium and in exudates. PMID- 11911188 TI - Characterization of the outermembrane proteins of Vibrio cholerae expressed in in vivo culture. AB - Two outer membrane proteins (Omps) of Vibrio cholerae O1, expressed in the intestine (in vivo) but not in culture media (in vitro), were investigated. The molecular masses of those proteins were 116 kDa and 15 kDa, and they were not associated with iron-regulated proteins. Convalescent cholera patients' sera reacted with the 15 kDa protein but not with the 116 kDa protein. The N-terminal amino acid sequence of the 15 kDa protein was homologous to V. cholerae OmpT. Anti-serum to the 15 kDa protein caused agglutination of the organisms grown in the intestine, but not the organisms in in vitro culture. The anti-serum was bactericidal, but it did not inhibit the adhesion of the organisms to the intestine and HEp-2 cells. These findings suggest the possibility that the 15 kDa protein could be involved in post-infection immunity. PMID- 11911189 TI - Cross-reactivity and sensitivity of two Legionella urinary antigen kits, Biotest EIA and Binax NOW, to extracted antigens from various serogroups of L. pneumophila and other Legionella species. AB - Legionella antigen detection kits for diagnosing legionellosis from urine have become widely used, but basic information about reactivity of the kits to non serogroup (SG) 1 L. pneumophila and other Legionella species remains incomplete. We evaluated Biotest EIA and the most recently developed Binax NOW by using in vitro extracted antigens of 22 L. pneumophila SG 1 to 15 strains and of 27 other Legionella species. Both kits showed excellent sensitivity to L pneumophila SG 1 antigens, but reacted to different sets of non-SG I L. pneumophila with different sensitivity. No cross-reactivity was observed to Legionella species other than L. pneumophila. PMID- 11911190 TI - Oral delivery of antigens in liposomes with some lipid compositions modulates oral tolerance to the antigens. AB - Several liposomes containing ovalbumin (OVA), a model antigen, with different lipid compositions were prepared in order to evaluate their ability to induce oral tolerance. Oral administration of these liposomal OVAs induced suppression of the proliferative responses of popliteal lymph node cells from the treated mice to OVA, suggesting that these treated mice were tolerized. The efficiency of the induction of oral tolerance was affected by the liposome composition. OVA entrapment in these liposomes could modulate the tolerizing dose of OVA itself. These results suggest that some liposomes can be suitable antigen-delivery systems for modulated and/or effective induction of oral tolerance. PMID- 11911192 TI - Effect of phosphatidylserine content on the partition coefficients of diazepam and flurazepam between phosphatidylcholine-phosphatidylserine bilayer of small unilamellar vesicles and water studied by second derivative spectrophotometry. AB - The affinity of the psychotropic benzodiazepine drugs diazepam (DZ) and flurazepam (FZ) to phosphatidylserine (PS) was examined since PS is abundantly contained in brain membranes. The effect of PS content on the partition coefficients (K(p)s) of these drugs between phosphatidylcholine (PC)-PS bilayer membranes of small unilamellar vesicles (SUV) and water was measured using second derivative spectrophotometry. The second derivative spectra of DZ and FZ measured in the solutions containing various amounts of PC-PS SUV clearly showed derivative isosbestic points and a distinct derivative intensity change depending on the amount of the SUV added. The derivative intensity differences (AD) of the drugs before and after addition of the SUV suspension were measured at a specific wavelength. Using the AD values, the Kp values were calculated and obtained with relative standard deviation of below 10%. The Kp values of both drugs increased according to the PS content in the PS-PC bilayer membranes of the SUV proving that both have higher affinity to the PC-PS bilayer membranes than to PC membranes. The effect was much larger for FZ, i.e., the Kp value of FZ at 30 mol% PS content increased to about five times the value for the PC SUV. This can be explained by the fact that at the experimental pH of 7.4, 80% of FZ molecules are in a cationic form (pKa=8.1), so that these molecules are highly accessible to the negatively charged PS molecules. The results support the rapid and high distribution of DZ and FZ in the central nervous system after their administration. PMID- 11911191 TI - Mushroom tyrosinase inhibition activity of some chromones. AB - Currently, aloesin is used in the cosmetic industry as a whitening agent because it inhibits tyrosinase activity. Aloesin is a C-glycosylated chromone compound isolated from aloe, and it is difficult to synthesize because of C-glycosyl moiety in the molecule. The purpose of this study is to search for a new chromone compound which is easy to synthesize and which posesses stronger tyrosinase inhibitory activity than aloesin. Fourteen chromone derivatives were synthesized and screened for their mushroom-tyrosinase inhibitory activity. 5-Methyl-7 methoxy-2-(2'-benzyl-3'-oxobutyl)chromone (15) showed the strongest activity among tested compounds. Its activity was not only stronger than aloesin, but also stronger than arbutin and kojic acid. The kinetic analysis revealed a competitive inhibition of 15 with tyrosinase for the L-tyrosine binding site. PMID- 11911193 TI - Structure activity relationships of new inhibitors of mammalian 2,3-oxidosqualene cyclase designed from isoquinoline derivatives. AB - We have designed more potent inhibitors from the previously reported LF 05-0038, a 6-isoquinolinol based inhibitor of 2,3-oxidosqualene cyclase (IC50: 1.1 microM). Replacement of the 3-OH group by various 3-substituted amino groups, and modification of the alkyl chain borne by the endocyclic nitrogen led to inhibitors with IC50 in the range of 0.15 to 1 microM. In a second step, opening of the bicyclic ring system afforded the corresponding aminoalkylpiperidines which were slightly more potent. Finally, introduction of suitable aromatic containing moieties on the piperidine nitrogen yielded very potent inhibitors such as 20x (IC50 = 18 nM) easy to synthesize and achiral. The recent availability of the crystal structure of squalene-hopene cyclase allowed us to construct a three-dimensional (3D) model of the related 2,3-oxidosqualene cyclase (OSC) which was tentatively used to describe the possible mode of binding of our compounds and which can be useful for designing new inhibitors. PMID- 11911194 TI - Five new chromones possessing monoamine oxidase inhibitory activity from an ascomycete, Chaetomium quadrangulatum. AB - Five novel chromones (1,4-benzopyran-4-ones), among which three are tetracyclic and one contains a sulfonyl group, have been isolated from an Ascomycete, Chaetomium quadrangulatum, as monoamine oxidase inhibitory features, and named chaetoquadrins A (1)-E (5). PMID- 11911195 TI - Potential insulinominetic agents of zinc(II) complexes with picolinamide derivatives: preparations of complexes, in vitro and in vivo studies. AB - Following the finding of in vitro insulinominetic activities of new prepared Zn(II) complexes with amide ligands (2-picolinamide (pa-a) and 6-methyl-2 picolinmethylamide (6mpa-ma)) in isolated rat adipocytes treated with epinephrine in terms of inhibition of free fatty acid release, their blood glucose normalizing effects were observed on daily intraperitoneal injections for 14 d in a type 2 diabetes mellitus model animal, KK-Ay mice. The blood glucose levels of KK-Ay mice were maintained in a normal range during the administration of both complexes. After the administration of each complex for 14 d, the improvement of glucose metabolism was confirmed as judged by the glucose tolerance test. PMID- 11911196 TI - Development of a novel vertical high shear kneader and its performance in wet kneading of pharmaceutical powders. AB - A novel multi-functional vertical high shear kneader has been developed. Wet kneading of pharmaceutical powders was conducted under various blade components and operating conditions. Compression properties of wet kneaded mass was analyzed and dispersion of hinder liquid (water) among the mass was investigated by assaying tracer aqueous pigment. Pellets were produced through a dome type extrusion granulator with continuous extrusion pressure measurement device and a fluidized bed drier, and then the physical properties were measured. Quantitative relationship between the pellet's physical properties and the binder dispersion condition as well as the compression properties could be obtained. It was found that the newly developed kneader was very effective to uniformly disperse binder as well as impart high shear stress to the wet mass without generating obvious adhesion onto the vessel wall. It was also pointed out that the extrusion pressure could determine the physical strength of pellet. This method proposes a new methodology for continuous monitoring of kneading condition as well as predicting pellet's physical properties. PMID- 11911197 TI - Efficient synthesis and hydrolysis of cyclic oxalate esters of glycols. AB - Based on the mechanism postulated for the formation of the cyclic carbonates 3 in the reactions of glycols 1 with oxalyl chloride in the presence of triethylamine, we present here three efficient syntheses of the cyclic oxalates 2 of various glycols 1 by controlling the formation of 3: replacement of the base by pyridine markedly diminishes yields of 3 in all reactions, realizing dramatic reversals of the product ratios in the reactions with the (R*,R*)-compounds 1g-i,q,r and pinacol (1k); although considerable amounts of the oxalate polymers are formed in the reactions with some (R*,S*)-glycols, this drawback can be removed by the use of 2,4,6-collidine instead of pyridine; 1,1'-oxalyldiimidazole is useful for the synthesis of two selected cyclic oxalates 2e,f. The cyclic oxalates 2 other than trisubstituted and tetrasubstituted ones were found to be very reactive: kinetic studies on the hydrolysis of 1,4-dioxane-2,3-dione (2a) as well as its mono- and some selected 5,6-disubstituted derivatives 2 have revealed that they undergo hydrolysis 260-1500 times more rapidly than diethyl oxalate (12) in acetate buffer-acetonitrile (pH 5.69) at 25 degrees C. Although the cyclic oxalate 21 from cis-1,2-cyclopentanediol (11) was 1.5 times more reactive than 2a, it has been shown with other substrates that increasing number of the alkyl substituents decreases the rate of hydrolysis. On the contrary, the phenyl group was found to have somewhat accelerative effect. PMID- 11911198 TI - Pharmacokinetic study of allixin, a phytoalexin produced by garlic. AB - The pharmacokinetic behavior of allixin (3-hydroxy-5-methoxy-6-methyl-2-penthyl 4H-pyran-4-one) was investigated in an experimental animal, mice. Allixin was administered using an inclusion compound because the solubility of allixin in aqueous solution is very low. The allixin content in serum and in the organs of administered animals was analyzed by liquid chromatography (LC)-MS. Most of the administered allixin disappeared within 2 h, and the bioavailability of allixin was estimated to be 31% by obtained area under the blood concentration-time curve (AUC). The metabolites of allixin were studied using the metabolic enzyme fraction of liver and liver homogenate. Several new peaks corresponding to allixin metabolites were observed in the HPLC chromatoprofile. The chemical structure of the metabolites was investigated using LC-MS and NMR. Three of them were identified as allixin metabolites having a hydroxylated pentyl group. PMID- 11911199 TI - Facile synthesis of 9-substituted 9-deazapurines as potential purine nucleoside phosphorylase inhibitors. AB - A facile synthesis of 9-substituted 9-deazapurines as potential inhibitors of purine nucleoside phosphorylase has been achieved by the direct Friedel-Crafts aroylation or arylmethylation of 9-deazapurines using trifluoromethanesulfonic acid as catalyst. The aroylated 9-deazapurines could be transformed into the corresponding 9-aryimethyl derivatives by the Wolff-Kishner reaction. A novel synthesis of 9-deazahypoxanthine was also developed by treatment of 4-hydroxy-5 phenylazo-6-methylpyrimidin-2-thione with triethyl orthoformate in trifluoroacetic acid (TFA) to yield 8-oxo-7H-2-phenylpyrimido[5,4-c]pyridazin-6 thione followed by Raney nickel reduction. PMID- 11911200 TI - Seven-membered vibsane-type diterpenes with a 5,10-cis relationship from Viburnum awabuki. AB - New five seven-membered vibsane-type diterpenes named 5-epi-vibsanin C, 5-epi vibsanin H, 5-epi-vibsanin K, 18-O-methyl-5-epi-vibsanin K and 5-epi-vibsanin E have been isolated from the leaves of Viburnum awabuki (Caplifoliaceae). Their structures have been elucidated by analyses of spectroscopic data and comparison of their spectral data with those of the previously known seven-membered vibsane type diterpenes. The occurrence of these seven-membered vibsane-type diterpenes with a cis relationship on the C-5 and C-10 positions in nature have been predicted by conformational analysis of vibsanin B, an eleven-membered vibsane type diterpene. Vibsanin C, 5-epi-vibsanin C and 5-epi-vibsanin H exhibited moderate cytotoxic activities on KB cells. PMID- 11911201 TI - Lewis acid-catalyzed asymmetric diels-alder reactions using chiral sulfoxide ligands: chiral 2-(arylsulfinylmethyl)-1,3-oxazoline derivatives. AB - New chiral sulfoxide-1,3-oxazoline ligands have been developed as chiral ligands for Lewis acid-catalyzed asymmetric Diels-Alder reactions. The use of chiral sulfinyl 1,3-oxazoline ligands in copper(II)-catalyzed asymmetric Diels-Alder reactions provided an endo cycloadduct as a major product with moderate enantioselectivity. A rationale is proposed for the mechanism of the asymmetric induction. PMID- 11911202 TI - Rare earth metal trifluoromethanesulfonates catalyzed benzyl-etherification. AB - Rare earth metal trifluoromethanesulfonates [rare earth metal triflate, RE(OTf)3] were found to be efficient catalyst for benzyl-etherification. In the presence of a catalytic amount of RE(OTf)3, condensation of benzyl alcohols and aliphatic alcohols proceeded smoothly to afford the benzyl ethers. The condensation between benzyl alcohols and thiols also proceeded, and thio ethers were obtained in good yield. In these reactions, RE(OTf)3 could be recovered easily after the reactions were completed and could be reused without loss of activity. PMID- 11911203 TI - Nine new secoiridoid glucosides from Jasminum nudiflorum. AB - Phytochemical study of the leaves of Jasminum nudiflorum has led to the isolation of nine new secoiridoid glucosides, jasnudiflosides F-L (1-7), nudifloside D (8) and isooleoacteoside (9). The structures of these compounds were elucidated on the basis of chemical and spectroscopic evidence. PMID- 11911204 TI - Platanionosides D-J: megastigmane glycosides from the leaves of Alangium platanifolium (Sieb. et Zucc.) Harms var. platanifolium Sieb. et Zucc. AB - From the leaves of Alangium paltanifolium var. platanifolium, collected in Fukuoka Prefecture, twelve further megastigmane glycosides were isolated. Seven of them, named platanionosides D-J (1-7), were found to be new compounds. Their structures were elucidated from spectroscopic evidence and their absolute structures were determined from beta-D-glucosylation-induced shift trends of 13C NMR and by application of a modified Mosher's method. PMID- 11911205 TI - Influence of pH on the binding of diphenylmethylenepiperidines by 5-HT2B receptors in rat stomach fundus. AB - Cyproheptadine is one of the compounds exhibiting the highest activity at 5-HT2B receptors. In a previous work we analysed the relevance of the amino group in diphenylmethylenepiperidines (DPMP), which are open cyproheptadine analogues. Only compounds containing N-H or N-methyl motifs, showed significant 5-HT2B activity. Surprisingly, the corresponding quaternary ammonium salt demonstrated a total lack of activity. Therefore, the question arises whether protonation favours the interaction of these compounds with 5-HT2B receptors. Consequently, we studied the protonation influence (by varying the pH of the medium) on the antagonism of serotonin by some cyproheptadine analogues in rat stomach fundus. The main results were: 1) N-protonation increases the activity of DPMPs. 2) Alkaline pH facilitates the occurrence of a non-surmountable antagonism. 3) The contrast between the activity of protonated DPMPs and the lack of activity of the corresponding quaternary ammonium cation, suggests either that the latter is prevented from acting by steric hindrance, or that the mechanism by which protonation may increase the activity depends not only on the charge of the proton, but also on its ability to form hydrogen bonds. PMID- 11911206 TI - Synthesis and insecticidal activity of novel 4beta-halogenated benzoylamino podophyllotoxins against Pieris rapae Linnaeus. AB - Twelve new 4beta-halogenated benzoylamino compounds (7.1-7.12) of podophyllotoxin have been synthesized, and their structures were confirmed by IR, 1H-NMR, MS spectra as well as CHN elemental analysis. These compounds showed delayed insecticidal activity against 5th instar larvae of Pieris rapae Linnaeus in vivo, when tested by a leaf-dipping method at a concentration of 250ppm. By preliminary qualitative structure-activity relationship analysis, we found the following results: 1) Compounds 7.2, 7.5-7.9 were more potent than the nature parent product in the mortality after 15 d against P. rapae in vivo. Especially compounds 7.5 and 7.6 bearing meta- and para-chlorobenzene substituents respectively, were the most potent of these compounds; 2) Substitution on the benzene ring moiety of 4beta-benzoylamino podophyllotoxin (PPT) with Cl, Br, I at the para or at the meta position yielded compounds which were as potent or more potent than those containing the corresponding substituting group at the ortho position. 3) Substitution on the benzene ring moiety of 4beta-benzoylamino podophyllotoxin with I either at the ortho, meta or para position yielded less potent compounds (7.10-7.12) when compared with PPT. PMID- 11911207 TI - A new isoflavone glycoside from Ceiba pentandra (L.) Gaertner. AB - From the 80% EtOH extract of the bark of Ceiba pentandra (L.) Gaertner, a new isoflavone glycoside was isolated along with known isoflavones, vavain and vavain glucoside. The structure was elucidated by spectroscopic analysis as 5-hydroxy 7,4',5'-trimethoxyisoflavone 3'-O-alpha-L-arabinofuranosyl(1-->6)-beta-D glucopyranoside. PMID- 11911208 TI - Allixin accumulation with long-term storage of garlic. AB - Extremely high accumulation of allxiin, a phytoalexin derived from garlic, was observed in necrotic tissue areas after long-term storage. The allixin produced recrystallized on the surface of the garlic clove. The amount of allixin produced in raw garlic with necrotic tissue areas was 1400 ng/mg wet garlic, which exceeds the minimum exhibitory concentration of allixin. After approximately 2 years of storage, amount of allixin accumulated reached slightly less than 1% of the dry weight of garlic cloves. PMID- 11911209 TI - Application of eudragit RS to thermo-sensitive drug delivery systems. I. Thermo sensitive drug release from acetaminophen matrix tablets consisting of eudragit RS/PEG 400 blend polymers. AB - In order to develop the polymer materials having temperature-sensitive and high biological safety, Eudragit RS-PO and polyethylene glycol 400 (PEG 400) blend polymers (EPG) were prepared. The EPGs that have the glass transition temperature (Tg) at around the body temperature were prepared by the addition of 5--13% PEG 400 to Eudragit RS. As glassy polymers are not in thermodynamic equilibrium below their Tg, the effects of isothermal aging on the T(g)s of Eudragit RS and EPG containing 10% PEG 400 (10% EPG) were also studied at various aging temperatures. The Tg values of Eudragit RS increased with the aging time and after 30 d of aging, they apparently reached constant values which markedly differed depending on the aging temperatures. On the other hand, the Tg values of 10% EPG were almost independent of the aging temperature and reached around 33 degrees C at 30 d after aging. The ability as thermo-sensitive polymer of EPG was evaluated by the dissolution test of the acetaminophen (AAP) matrix tablets prepared with EPG. The AAP release rate from the EPG matrix tablets slightly changed below the Tg of tablets, and then, it markedly increased above the Tg. Considering high biological safety of Eudragit RS and PEG 400, EPG might be available to develop the novel thermo-sensitive drug delivery systems. PMID- 11911210 TI - Megistoquinones I and II, two quinoline alkaloids with antibacterial activity from the bark of Sarcomelicope megistophylla. AB - Two alkaloids, megistoquinone I (1) and megistoquinone II (2), were isolated from the bark of Sarcomelicope megistophylla. Their structures have been elucidated on the basis of MS and NMR data. Both belong to quinoline alkaloid series and should be considered as oxidation products of a furo[2,3-b]quinoline precursor. The two alkaloids showed antibacterial properties with minimum inhibitory concentration (MIC) ranging from 2.35 to 5.25 mg/ml for 1 and 0.73 to 1.23 mg/ml for 2. PMID- 11911211 TI - Syntheses of 1beta-methylcarbapenems bearing 5-methyl-4-hydroxypyrrolidinone. AB - A new series of 1beta-methylcarbapenems 1a-d bearing 5-methyl-4 mercaptopyrrolidinone rings has been prepared and evaluated for in vitro antibacterial activity and pharmacokinetic parameters. Most compounds showed excellent antibacterial activity and high stability to dehydropeptidase-1. We have synthesized optically active 5-methyl-4-hydroxypyrrolidinones from enantiomerically pure aziridine esters. PMID- 11911212 TI - Six new 2-(2-phenylethyl)chromones from Agarwood. AB - Six new chromones, 6-methoxy-2-[2-(3-methoxy-4-hydroxyphenyl)ethyllchromone (2), 6,8-dihydroxy-2-(2-phenylethyl)chromone (3), 6-hydroxy-2-[2-(4 hydroxyphenyl)ethyl]chromone (4), 6-hydroxy-2-[2-(2-hydroxyphenyl)ethyl]chromone (5), 7-hydroxy-2-(2-phenylethyl)chromone (6), and 6-hydroxy-7-methoxy-2-(2 phenylethyl)chromone (7) were isolated from the ether extract of agarwood in addition to a known compound, 2-(2-phenylethyl)chromone or flidersiachromone (1). Their structures were determined by spectroscopic methods including UV, IR, and NMR spectral data and comparisons with the calculated values using the hydroxyl and methoxyl substituent increments of the chromone ring. PMID- 11911213 TI - Synthesis of the side chain of a novel carbapenem via iodine-mediated oxidative cyclization of (1R)-N-(1-aryl-3-butenyl)acetamide. AB - A (2R,4S)-trans-disubstituted pyrrolidine ring system was constructed by employing iodine-mediated oxidative cyclization of (1R)-N-[1-(4-bromophenyl)-3 butenyl]acetamide 3 as a key step. The resulting diastereomeric mixture of (2R)-2 aryl-4-acetoxypyrrolidine 4 was stereoselectively converted to the side-chain of a novel ultrabroad-spectrum carbapenem 1, via (2R,4R)-2-aryl-4-hydroxypyrrolidine 7. PMID- 11911214 TI - Novel quinazoline ring synthesis by cycloaddition of N-arylketenimines with N,N disubstituted cyanamides. AB - The reaction of N-aryl-substituted ketenimines with N,N-disubstituted cyanamides or (MeS)2C=N-CN under high pressure afforded 4-(N,N-disubstituted amino) or 4 (MeS)2C=N-substituted quinazoline derivatives, respectively. These products were formed by [4+2] cycloaddition between the aza-diene moieties of the N arylsubstituted ketenimines and cyano groups. A 4-(unsubstituted amino)quinazoline derivative was synthesized by hydrolysis of the latter product. PMID- 11911216 TI - Facile formation of chiral calixarene analogs incorporating cystine peptide into the macrocyclic ring. AB - Chiral calixarene analogs incorporating cystine peptide into their macrocyclic ring were easily prepared by the cyclization reactions of bis(chloromethyl)phenol formaldehyde oligomers with cystine peptides in moderate yields. Circular dichroism (CD) spectra indicated the existence of the transmission of the chirality from peptide unit to phenol-formaldehyde oligomer moiety. PMID- 11911215 TI - Antidiabetic principles of natural medicines. V. Aldose reductase inhibitors from Myrcia multiflora DC. (2): Structures of myrciacitrins III, IV, and V. AB - Following the characterization of myrciacitrins I and II and myrciaphenones A and B, three new flavanone glucosides, myrciacitrins III, IV, and V, were isolated from the leaves of Brazilian Myrcia multiflora. The structures of new myrciacitrins were elucidated on the basis of physicochemical and chemical evidence. Myrciacitrins were found to show potent inhibitory activity on aldose reductase. PMID- 11911217 TI - Highly accelerating effect of lewis acids on ruthenium(II)-catalyzed radical addition reactions. AB - The intramolecular ruthenium(II)-catalyzed radical addition of the trichloroacetyl pendant group to the 2-oxazolone skeleton is greatly enhanced in the presence of catalytic Lewis acids including rare earth metal triflates, thus providing a convenient route to a highly potential chiral synthon for vic-amino alcohols. PMID- 11911218 TI - Two novel long-chain alkanoic acid esters of lupeol from alecrim-propolis. AB - Two new long-chain alkanoic acid esters of lupeol were isolated together with known triterpenoids, alpha-amyrin, beta-amyrin, cycloartenol, lanosta-7,24-diene 3beta-ol and lupeol from Alecrim-propolis collected in Brazil. The structures were characterized by spectroscopic means. PMID- 11911219 TI - New entry of coupling reaction of phenacylamine derivatives with silylstannane. AB - The new coupling reaction of phenacylamines with silylstannane and lithium diisopropylamide (LDA) is reported. The treatment of a phenacylamine iodide 1 with (trimethylsilyl)tributylstannane (Me3SiSnBu3) and cesium fluoride (CsF) gave a dimerization product 2 having no iodine atom. Reaction of 1 with LDA afforded a dimerization product 3 with an iodine atom. The products 2 and 3 were separated to the meso and racemic isomers, respectively. PMID- 11911220 TI - Effect of vitamin A deficiency on cardiovascular function in the rat. AB - Selected parameters of cardiovascular function were evaluated in vitamin A deficient rats at 70 days of age. Resting heart rate was increased by an average of 100 bpm (21.4+/-2.7%), whereas resting systolic blood pressure was normal in vitamin A-deficient animals. The maximal contractile force developed per milligram weight of tissue by aortic rings excised from vitamin A-deficient animals was reduced in response to high potassium (-25.0+/-8.7%) and phorbol 12,13-dibutyrate (-36.8+/-8.4%) but was only slightly reduced in response to norepinephrine (-17.8+/-11.1%). Intimal rubbing to remove the endothelium had no effect on the loss in contractile responsiveness, and the relaxant response to acetylcholine was similar between control and vitamin A-deficient tissue groups. This suggests that the decrease in contractility of vascular smooth muscle from the vitamin A-deficient rats did not involve altered release of endothelium derived vasoactive factors. Western blot analysis suggested a reduction in the protein levels of several differentiation markers including alpha-actin (-22%), calponin (-37%), desmin (-37%), and vinculin (-40%), whereas the level of PKCalpha was unchanged from control values. Our findings indicate a significant decrease in contractile responsiveness of aortic smooth muscle of the vitamin A deficient rat that may be associated with a down regulation in the expression of contractile-related proteins. PMID- 11911221 TI - Inhibition of rat lipoprotein lipid peroxidation by the oral administration of D003, a mixture of very long-chain saturated fatty acids. AB - Previous results have demonstrated that policosanol, a mixture of aliphatic primary alcohols isolated and purified from sugar cane wax, whose main component is octacosanol, inhibited lipid peroxidation in experimental models and human beings. D003 is a defined mixture of very long-chain saturated fatty acids, also isolated and purified from sugar cane wax, whose main component is octacosanoic acid followed by traicontanoic, dotriacontanoic, and tetracontanoic acids. Since very long-chain fatty acids are structurally related to their corresponding alcohols, we investigated the effect of oral treatment with D003 (0.5, 5, 50, and 100 mg/kg) over 4 weeks in reducing the susceptibility of rat lipoprotein to oxidative modification. The combined rat lipoprotein fraction VLDL + LDL was subjected to several oxidation systems, including those containing metal ions (CuSO4), those having the capacity to generate free radicals 2,2-azobis-2 amidinopropane hydrochloride (AAPH), and a more physiological system (resident macrophages). D003 (5, 50, and 100 mg/kg) significantly inhibited copper-mediated conjugated-diene generation in a concentration-dependent manner. D003 increased lag phase by 53.1, 115.3, and 119.3%, respectively, and decreased the rate of conjugate-diene generation by 16.6, 21.5, and 19.6%, respectively. D003 also inhibited azo-compound initiated and macrophage-mediated lipid peroxidation as judged by the significant decrease in thiobarbituric acid reactive substance (TBARS) generation. In all the systems the maximum effect was attained at 50 mg/kg. There was also a parallel attenuation in the reduction of lysine amino groups and a significant reduction of carbonyl content after oxidation of lipoprotein samples. Taken together, the present results indicate that oral administration of D003 protects lipoprotein fractions against lipid peroxidation in the lipid as well in the protein moiety. PMID- 11911222 TI - Combination IK1 and IKr channel blockade: no additive lowering of the defibrillation threshold. AB - Selective blockade of the inward rectifier potassium channel I(K1) by barium, or of the rapidly activating delayed rectifier potassium channel I(Kr) by D,L sotalol, prolongs repolarization and reduces the defibrillation threshold (DFT). This study hypothesized that combination I(K1) and I(Kr) channel block would produce concentration-dependent additive effects on DFT and ventricular refractoriness. A range of barium and D,L-sotalol concentrations, alone and in combination, were examined with respect to DFT, ventricular effective refractory period (VERP), and ventricular fibrillation cycle length (VFCL) in 133 Langendorff-perfused rabbit hearts. Barium produced a concentration-dependent reduction of DFT (-49+/-4%), with concentration-dependent increases in VERP (26+/ 6%) and VFCL (42+/-18%). D,L-Sotalol produced a concentration-dependent lowering of DFT (-53+/-6%) with a concentration-dependent increase in VFCL (34+/-8%) but not VERP. Low (1.6 microM), intermediate (3.1 microM), and high (12.5 microM) barium concentrations combined with varying D,L-sotalol concentrations produced equal or smaller decreases in DFT compared with corresponding doses of barium or D,L-sotalol alone. Except at the lowest concentrations of barium (1.6 and 3.1 microM) (p < 0.05), there was no significant additive interaction between barium and D,L-sotalol on VERP or VFCL. Combination I(K1) and I(Kr) channel block by barium and D,L-sotalol does not produce additive reduction of DFT. PMID- 11911223 TI - [Nphe1]nociceptin(1-13)-NH2 antagonizes nociceptin-induced hypotension, bradycardia, and hindquarters vasodilation in the anesthetized rat. AB - The purpose of this study was to investigate the effects of [Nphe1]nociceptin(1 13)-NH2 on nociceptin-induced decreases in mean arterial pressure (MAP), heart rate (HR), and hindquarters vascular bed resistance (HVBR) of the anesthetized rat. The results showed that i.c.v. or i.v. [Nphe1]nociceptin(1-13)-NH2 (1.5-12 nmol/kg and 5-120 nmol/kg, respectively) could antagonize the depressor effects of i.c.v. or i.v. nociceptin (3 and 30 nmol/kg, respectively) on MAP and HR. Furthermore, [Nphe1]nociceptin(1-13)-NH2 (5-120 nmol/kg) could reverse nociceptin (30 nmol/kg)-induced decrease of HVBR. However, [Nphe1]nociceptin(1-13)-NH2 had no significant effects on similar effects induced by morphine. Our results suggest that [Nphe1]nociceptin(1-13)-NH2 acts as a selective antagonist of the nociceptin receptor in the cardiovascular system of the rat. PMID- 11911224 TI - No change in dopamine D1 receptor in vivo binding in rats after sub-chronic haloperidol treatment. AB - A frequent side effect in the long-term treatment of schizophrenia with the dopamine D2 antagonist haloperidol (HAL) is the appearance of tardive dyskinesia or, in animals, of repetitive involuntary vacuous chewing movements (VCMs). In rats, chronic HAL-induced or D1 receptor-stimulated VCMs are suppressed by D1 antagonists, suggesting that this behavioral supersensitivity is mediated by D1 receptors. The goal of this study was to investigate in vivo the possible relationship between D1 receptor binding and D1-mediated behavioral supersensitivity, after subchronic HAL treatments. D1 agonist R-SKF 82957 and antagonist SCH 23390, both labeled with carbon-11, were used to assess in vivo D1 receptor binding. Rats were treated with HAL (1.5 mg/kg, i.p.) or vehicle for 21 days, followed by a 4 day washout period. No significant difference was found in the regional brain binding of either radioligand. D1 receptor-mediated behaviors including VCMs, grooming, and rearing were measured in control or HAL-treated rats. VCMs were significantly increased in HAL-treated rats, suggesting D1 receptor stimulation and possibly receptor supersensitivity. This study failed to link the purported D1 receptor-mediated behaviors with in vivo receptor binding measures of R-[11C]SKF 82957 or [11C]SCH 23390 in rat brain regions. PMID- 11911225 TI - Kinetic characterization and inhibition of the rat MAB elastase-2, an angiotensin I-converting serine protease. AB - An elastase-2 has been recently described as the major angiotensin (Ang) II forming enzyme of the rat mesenteric arterial bed (MAB) perfusate. Here, we have investigated the interaction of affinity-purified rat MAB elastase-2 with some substrates and inhibitors of both pancreatic elastases-2 and Ang II-forming chymases. The Ang II precursor [Pro 11 -D-Ala 12]-Ang I was converted into Ang II by the rat MAB elastase-2 with catalytic efficiency of 8.6 min-1 microM-1, and the chromogenic substrates N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide and N succinyl-Ala-Ala-Pro-Phe-p-nitroanilide were hydrolyzed by the enzyme with catalytic efficiencies of 10.6 min-1 microM-1 and 7.6 min-1 microM-1, respectively. The non-cleavable peptide inhibitor CH-5450 inhibited the rat MAB elastase-2 activities toward the substrates Ang I (IC50 = 49 microM) and N succinly-Ala-Ala-Pro-Phe-p-nitroanilide (IC 50 = 4.8 microM), whereas N-acetyl Ala-Ala-Pro-Leu-chloromethylketone, an effective active site-directed inhibitor of pancreatic elastase-2, efficiently blocked the Ang II-generating activity of the rat MAB enzyme (IC 50 = 4.5 microM). Altogether, the data presented here confirm and extend the enzymological similarities between pancreatic elastase-2 and its rat MAB counterpart. Moreover, the thus far unrealized interaction of elastase-2 with [Pro 11-D-Ala 12]-Ang I and CH-5450, both regarded as selective for chymases, suggests that evidence for the in vivo formation of Ang II by chymases may have been overestimated in previous investigations of Ang II-forming pathways. PMID- 11911226 TI - Effects of captopril and omapatrilat on early post-myocardial infarction survival and cardiac hemodynamics in rats: interaction with cardiac cytokine expression. AB - The purpose of this study was to evaluate and compare the effects of simultaneous angiotensin-converting enzyme (ACE) and neutral endopeptidase 24.11 (NEP) inhibition by the vasopeptidase inhibitor omapatrilat (10 and 40 mg x kg(-1) x day(-1)) with those of the selective ACE inhibitor captopril (160 mg x kg(-1) x day(-1)) on survival, cardiac hemodynamics, and cytokine mRNA expression in left ventricular (LV) tissues 4 days after myocardial infarction (MI) in rats. The effects of the co-administration of both B1 and B2 kinin receptor antagonists (2.5 mg x kg(-1) x day(-1) each) with and without omapatrilat were also evaluated to assess the role of bradykinin (BK) during this post-MI period. Both omapatrilat and captopril treatments improve early (4 days) post-MI survival when started 4 h post-MI. The use of kinin receptor antagonists had no significant effect on survival in untreated MI rats and omapatrilat-treated MI rats. This improvement in survival with omapatrilat and captopril is accompanied by a reduced LV end-diastolic pressure (LVEDP) and pulmonary congestion. The use of kinin receptor antagonists had little effect on cardiac hemodynamics or morphologic measurements. Acute MI significantly increased the expression of cardiac cytokines (TNF-alpha, TGF-beta1, and IL-10). Captopril significantly attenuated this activation, while omapatrilat had variable effects: sometimes increasing but generally not changing activation depending on the cytokine measured and the dose of omapatrilat used. The co-administration of both kinin receptor antagonists attenuates the increase in expression of cardiac TNF-alpha and TGF-beta1 after omapatrilat treatment. Taken together, these results would suggest that despite very marked differences in the way these drugs modified the expression of cardiac cytokines, both omapatrilat and captopril improved early (4 days) post-MI survival and cardiac function to a similar extent. PMID- 11911227 TI - Effect of cannabinoids on neural transmission in rat gastric fundus. AB - The purpose of this study was to examine the possible role of cannabinoids on the neuromuscular function of rat gastric fundus. In addition to possible direct effects on smooth muscle, the influence of cannabinoids on contractile (cholinergic) and relaxant (non-adrenergic, non-cholinergic (NANC)) neural innervation of the rat gastric fundus was investigated in vitro. Neither anandamide (an endogenous cannabinoid receptor agonist) nor Win 55,212-2 and methanandamide (synthetic cannabinoid receptor agonists) nor AM 630 (a cannabinoid receptor antagonist) showed any effect on smooth muscle activity at baseline or after precontraction with 5-hydroxytryptamine (5-HT; 10(-7) M). Electrical field stimulation (EFS) of the smooth muscle preparation (40 V; 5 Hz) caused cholinergically mediated twitch contractions that were abolished by atropine (10(-6) M) or tetrodotoxin (TTX; 10(-6) M). Anandamide and Win 55,212-2 reduced these twitch contractions in a concentration-dependent manner, an effect that could be reversed by the cannabinoid receptor antagonist AM 630 for anandamide, but not for Win 55,212-2. When NANC relaxant neural responses (presence of atropine (10(-6) M) and guanethidine (10(-6) M)) were induced by EFS, the cannabinoid receptor agonists anandamide and Win 55,212-2 reduced the relaxant response, an effect that could be reversed by the cannabinoid receptor antagonist AM 630 for anandamide, but not for Win 55,212-2. When given alone AM 630 caused an increase in the EFS-induced relaxant response. The presence of CB1 and CB2 cannabinoid receptor mRNA within the rat stomach was demonstrated by reverse transcription polymerase chain reaction (RT-PCR). The results of this study indicate that cannabinoids modulate excitatory cholinergic and inhibitory NANC neurotransmission in the rat gastric fundus. Endogenous cannabinoids may play a physiological role only in NANC inhibitory transmission, as AM 630 did not modify the electrically induced cholinergic contraction. The involved cannabinoid receptors are most likely located on neuronal structures. The present study also provides evidence that more than one receptor type is involved. PMID- 11911228 TI - The role of PI 3-kinase in EGF-stimulated jejunal glucose transport. AB - Epidermal growth factor (EGF) rapidly increases jejunal glucose transport. Signal transduction mechanisms mediating EGF-induced alterations in jejunal glucose transport remain to be determined. New Zealand White rabbit (1 kg) jejunal tissue was stripped and mounted in short-circuited Ussing chambers. The transport of tritiated 3-O-methylglucose was measured in the presence of the PKC agonist 1,2 dioctanoyl-sn-glycerol (1,2-DOG) or the inactive analog 1,3-dioctanoyl-sn glycerol (1,3-DOG). Additional experiments examined the effect of the PKC inhibitor chelerythrine, the PLC inhibitor U73122, the MAPK inhibitor PD 98059, the G-protein inhibitor GDP-betaS, the PI 3-kinase inhibitor LY294002, or the microtubule inhibitor colchicine on EGF-induced jejunal glucose transport. Net jejunal 3-O-methylglucose absorption was significantly increased following specific activation of PKC. A PKC antagonist inhibited the EGF-induced increase in net 3-O-methylglucose transport, while PI 3-kinase inhibition completely blocked the EGF-induced transport increase. Inhibition of PLC, MAPK, G-proteins, and microtubules had no effect on EGF-stimulated increases in jejunal transport. We conclude that the effect of EGF on jejunal glucose transport is mediated at least in part by PKC and PI 3-kinase. PMID- 11911229 TI - Insulin sensitivity regulated by feeding in the conscious unrestrained rat. AB - Hepatic insulin sensitizing substance (HISS), a putative hormone released from the liver in response to insulin in fed animals, accounts for 50-60% of insulin action. HISS release is regulated by permissive control of the hepatic parasympathetic nerves. The objectives were to develop the rapid insulin sensitivity test (RIST) in conscious rats, and to assess the effects of anesthesia, atropine, feeding, and fasting on insulin action. The RIST index, expressed as milligrams glucose per kilogram body weight required to maintain euglycemia after a 50 mU/kg bolus of insulin, was similar in conscious and anesthetized rats (238.6+/-42.5 vs. 225.3+/-30.4 mg/kg). Atropine produced a 56% inhibition of insulin action in fed rats. After a 24 h fast, full HISS-dependent insulin resistance had developed as shown by a low RIST index that was not reduced further by atropine. Fasting caused a 10.5% decrease in insulin action per hour over six hours. HISS-dependent insulin resistance in 24-h fasted rats was reversed 4 h after re-feeding (90.9+/-12.3 vs. 204.5+/-30.5 mg/kg). We conclude that HISS-dependent and HISS-independent insulin action, as assessed by the RIST, is similar in conscious and pentobarbital-anesthetized rats. Pharmacological blockade of HISS-dependent insulin action and physiological regulation of HISS action by feeding-fasting is confirmed. Re-feeding fasted rats reversed HISS-dependent insulin resistance. Merits of use of the RIST in conscious versus anesthetized rats are discussed. PMID- 11911230 TI - Validity of the Wechsler abbreviated scale of intelligence and other very short forms of estimating intellectual functioning. AB - Performance on the Wechsler Adult Intelligence Scale-III (WAIS-III) was compared to performance on the Wechsler Abbreviated Scale of Intelligence (WASI), as well as short form estimations of intellectual functioning derived from WAIS-III performance, in a mixed clinical sample of 72 participants. The WASI verbal IQ (VIQ) score was significantly higher than the WAIS-III VIQ, whereas performance IQ (PIQ) estimates all differed from actual WAIS-III PIQ and full scale IQ (FSIQ). Correlations of WAIS-III scores with WASI scores were consistently lower than were correlations between the WASI-III and all other short forms. Although maintaining administration times of 15 minutesfor a two-subtest FSIQ and 30 minutes for a four-subtest FSIQ, the WASI did not consistently demonstrate desirable accuracy in predicting scores obtained from the WAIS-III. The results suggest that clinicians should use the WASI cautiously if at all, especially when accurate estimates of individuals' WAIS-III results are needed. PMID- 11911231 TI - The Minnesota Multiphasic Personality Inventory-2 across the human immunodeficiency virus spectrum. AB - The Minnesota Multiphasic Personality Inventory-2 (MMPI-2) was used to assess individuals' patterns of psychological symptoms across the spectrum of HIV illness. Two hundred and twenty-five participants in the present sample were administered the MMPI-2, 61 were HIV-seronegative controls, 61 were asymptomatic, 36 were symptomatic, and 67 met criteria for AIDS. Symptomatic HIV-seropositive patients scored higher on the Hypochondriasis, Conversion-Hysteria, and Depression Scales. These differences appeared to be largely due to an increase in somatic complaints rather than an increase in other depressive symptoms. Group differences did not appear to be due to HIV-associated neuropsychological dysfunction. Interpretive strategies for the MMPI-2 and treatment considerations are discussed. PMID- 11911232 TI - Perinatal Bereavement Grief Scale: distinguishing grief from depression following miscarriage. AB - The study evaluated the Perinatal Bereavement Grief Scale (PBGS), the first scale designed to measure grieffollowing reproductive loss in terms of yearning for the lost pregnancy and lost baby. Participants included 304 women interviewed by telephone I to 3 times within 6 months aftermiscarriage. The PBGS had high internal consistency and test-retest reliability. It showed convergent validity with measures of attachment and investment in the child and divergent validity against measures of social desirability and depressive symptoms, supporting the conceptual distinction between grief and depression. Cross-cultural validity was acceptable whether tested by language (Spanish vs. English) or ethnicity (Hispanic vs. other). This measure of yearning enables study of the epidemiology and prognostic value of this key feature of mourning. PMID- 11911233 TI - A structure-based approach to psychological assessment: matching measurement models to latent structure. AB - The present article sets forth the argument that psychological assessment should be based on a construct's latent structure. The authors differentiate dimensional (continuous) and taxonic (categorical) structures at the latentand manifest levels and describe the advantages of matching the assessment approach to the latent structure of a construct. A proper match will decrease measurement error, increase statistical power, clarify statistical relationships, and facilitate the location of an efficient cutting score when applicable. Thus, individuals will be placed along a continuum or assigned to classes more accurately. The authors briefly review the methods by which latent structure can be determined and outline a structure-based approach to assessment that builds on dimensional scaling models, such as item response theory, while incorporating classification methods as appropriate. Finally, the authors empirically demonstrate the utility of their approach and discuss its compatibility with traditional assessment methods and with computerized adaptive testing. PMID- 11911234 TI - The daily inventory of stressful events: an interview-based approach for measuring daily stressors. AB - This study introduces the Daily Inventory of Stressful Events (DISE), an interview-based approach to the measurement of multiple aspects of daily stressors through daily telephone interviews. Using a U.S. national sample of adults aged 25 to 74 (N = 1,031), the prevalence as well as the affective and physical correlates of daily stressors are examined. Respondents had at least one daily stressor on 40 percent of the study days and multiple stressors on 11 percent of the study days. The most common class of stressors was interpersonal tension followed by work-related stressors for men and network stressors (events that occur to close others) for women. Stressors that involved danger of loss were more prevalent than stressors in which loss actually occurred. Regression analyses showed that specific types of daily stressors such as interpersonal tensions and network stressors were unique predictors of both health symptoms and mood. PMID- 11911235 TI - The use of reliable digits to detect malingering in a criminal forensic pretrial population. AB - The present research is a cross-validation of previous investigation by Greiffenstein, Baker; and Gola; Greiffenstein, Gola, and Baker; and Meyers and Volbrecht on the reliable digits (RELD) method of detecting suspected malingering on the Wechsler Adult Intelligence Scale-Revised (WAIS-R). The results support the use of the RELD method on a criminalforensic pretrial population (N = 187). Sensitivities, specificities, and incremental hit ratesfor two cut levels of the RELD method, Minnesota Multiphasic Personality Inventory-2 (MMPI-2) Infrequency and the Personality Assessment Inventory Negative Impression Scales, as well as multiple combined cut scores, were comparable to those observed in previous studies that used neuropsychologically evaluated participants. The selection of which cut score or combination of cut scores is appropriate on the RELD method is also discussed. PMID- 11911236 TI - Identifying faking bad on the Minnesota Multiphasic Personality Inventory Adolescent with Mexican adolescents. AB - This study examined the extent to which the validity scales of the Minnesota Multiphasic Personality Inventory-Adolescent identified Mexican adolescents who were instructed to fake bad. Validity scales data were used to differentiate between nonclinical adolescents instructed to fake bad and both clinical and nonclinical adolescents who received standard instructions. Participants were 59 male and 87 female Mexican high school students and 59 male and 87 female Mexican adolescents from clinical settings. This is the first study onfaking with adolescents in Mexico. The F, Fl, and F2 Scales and the F-K index discriminated adequately between the three different groups. Results were similar to those previously reportedfor adults and adolescents in Mexico and the United States. High positive and negative predictive powers and overall hit rates were obtained in this study. Higher cut scores were needed to discriminate between the groups of girls than between the groups of boys. PMID- 11911237 TI - Measures of self-efficacy and optimism in older adults with generalized anxiety. AB - This study provides initial psychometric data for the Self-Efficacy Scale (SES) and the Life Orientation Test (LOT) in a sample of older adults with generalized anxiety disorder (GAD). Participants included 76 adults, ages 60 to 80, who met Diagnostic and Statistical Manual of Mental Disorders (4th ed.) criteria for GAD. Self-efficacy and outcome expectancies were lower in older adults with GAD relative to published data from younger and older community samples. Both the SES and LOT demonstrated adequate internal consistency. Confirmatory factor analysis provided evidence for optimism and pessimism factors within the LOT and exploratory factor analysis of the SES suggested threefactors that overlap with previous findings. Overall, the data support the potential utility of these instruments in late-life GAD and set the stage forfuture investigations of generalized self-efficacy expectancies and outcome expectancies (or optimism) as they relate to the prediction of affect and behavior in this group. PMID- 11911238 TI - Factor and subtest discrepancies on the differential ability scales: examining prevalence and validity in predicting academic achievement. AB - Past literature has largely ignored the population frequency of multivariate factor and subtest score discrepancies. Another limitation has been that statistical models imperfectly model the clinical assessment process, whereby significant discrepancies between both factors and subtests are included in predictions about an individual's academic achievement. The present study examined these issues using a nationally representative sample (N = 1,185) completing the Differential Ability Scales. Results indicate that approximately 80% of children in a nonreferred sample show at least one statistically significant ability discrepancy. In addition, the global estimate of cognitive ability was the most parsimonious predictor of academic achievement, whereas information about ability discrepancies did not significantly improve prediction. Findings suggest that when predicting academic achievement, there is little value in interpreting cognitive scores beyond the global ability estimate. PMID- 11911239 TI - Confirmatory factor analysis of single- and multiple-factor competing models of the dissociative experiences scale in a nonclinical sample. AB - Previous research on the Dissociative Experiences Scale (DES) has demonstrated that (a) dissociation is quantifiable in both clinical and nonclinical samples and (b) a three-factor structure (amnesia, depersonalization, and absorption) is tenable for clinical samples. The factor structurefor nonclinical samples is less clear, with one- and multiple-factor solutions proposed. To clarify the DESfactor structure in nonclinical samples, confirmatory factor analyses were conducted on (a) one-, two-, three-, and four-factorfirst-order models and (b) two bifactor (hierarchical) models of DES scoresfor two samples of nonclinical university students. Results of delta(chi2) and goodness-of-fit indices support the three factor (first-order) model as bestfitting of the datafor these samples. The utility of this DES model for screening both dissociative pathology and elevated normal dissociative behavior in clinical and nonclinical populations is discussed. PMID- 11911240 TI - Interferon-gamma modulates TRAIL-mediated apoptosis in human colon carcinoma cells. AB - BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is expressed by immune cells and has been shown to play an important role in tumor surveillance due to its ability to induce apoptosis in various transformed cells. Interferon gamma (IFN-gamma), a multipotent cytokine with broad stimulatory effects on anti-tumoral immune reactions, may exert its cytotoxic activity directly on tumor cells or indirectly via stimulation of effector cells. This study was designed to determine the effect of IFN-gamma on TRAIL-mediated apoptosis in human colon carcinoma cell lines. MATERIALS AND METHODS: Cytotoxicity was assessed by MTT-assay. Expression of death receptors was measured by reverse transcriptase polymerase chain reaction. Apoptosis was assessed by caspase-8 immunoblot, DNA fragmentation and morphological studies. RESULTS: Treatment with TRAIL resulted in detectable cytotoxicity within 5 hours and was enhanced in a dose-dependent manner. When cells were pretreated with IFN gamma, the cytotoxic effect of TRAIL increased significantly. Treated cells showed a typical apoptotic morphology that was accompanied by internucleosomal cleavage of DNA. Up-regulation of caspase-8 expression and activation were detected as a result of pretreatment with IFN-gamma and subsequent apoptosis mediated by TRAIL. CONCLUSION: Our results demonstrated that IFN-gamma sensitises human colon carcinoma cells to TRAIL-mediated apoptosis, partly by elevated caspase-8 expression. PMID- 11911241 TI - Expression of progranulin and the epithelin/granulin precursor acrogranin correlates with neoplastic state in renal epithelium. AB - BACKGROUND: Current traditional pathological parameters, including staging and grading, are not sufficient in predicting outcome in patients with renal cell carcinoma (RCC). Acrogranin is an epithelial growth factor and has been demonstrated to play a role in teratocarcinogenesis and tumorigenesis. The aim of this study was to examine levels of acrogranin in renal cancer. MATERIALS AND METHODS: Western blot analysis was performed on renal tissue protein lysates. In addition, immunohistochemical (IHC) analysis of acrogranin expression was conducted on tissue sections of various histological types and grades of RCC. RESULTS: Western analysis showed that acrogranin levels were low in benign renal tissue and increased in malignant renal tissue. In addition, IHC revealed that high-grade RCC exhibited higher levels of expression than low-grade RCC and normal tissue. CONCLUSION: These data suggest that acrogranin may be a functional important growth factor in RCC and may be a potential molecular marker for high grade RCC. PMID- 11911242 TI - Chemopreventive effects of green tea polyphenols correlate with reversible induction of p57 expression. AB - Green tea polyphenols are known to induce apoptosis in certain types of tumor cells. However, the mechanism(s) that enables normal cells to evade the apoptotic effect is still not understood. In this study, Western blot analysis combined with cycloheximide treatment was used to examine the effects of green tea polyphenols on the expression levels of p57, a cyclin-dependent kinase and apoptosis inhibitor, in normal human keratinocytes and in the oral carcinoma cell lines SCC25 and OSC2. The results showed that the most potent green tea polyphenol, (-)-epigallocatechin-3-gallate (EGCG), induced p57 in normal keratinocytes in a dosage- and time-dependent manner, while the levels of p57 protein in oral carcinoma cells were unaltered. The differential response in p57 induction was consistent with the apoptosis status detected by annexin V assay. The data suggest that the chemopreventive effects of green tea polyphenols may involve p57-mediated cell cycle regulation in normal epithelial cells. PMID- 11911243 TI - The neurotrophin receptor p75NTR is a tumor suppressor in human prostate cancer. AB - The loss of tumor suppressor gene function contributes to the transformation of human prostate epithelial cells to a malignant pathology. Previous studies have demonstrated a loss of protein expression of the neurotrophin receptor, the p75NTR, from the pathological human prostate. Through cell cycle analysis, we demonstrate that a dose-dependent increase in the expression of p75NTR protein induces increased quiescence of a series of prostate tumor cells in vitro. When the same series of tumor cells were injected into the flanks of SCID mice, the growth of prostate tumors was suppressed in proportion to increased p75NTR expression. Tumor analysis showed that a dose-dependent increase in p75NTR expression was associated with a dose-dependent increase in the proportion of apoptotic cells, as determined by TUNEL assay, and a dose-dependent decrease in the proportion of cells committed to proliferation, as determined by proliferating cell nuclear antigen assay. These results demonstrate that the neurotrophin receptor p75NTR is a tumor suppressor of prostate growth by enhancing cell cycle quiescence, reducing proliferation and increasing apoptosis of the tumor cells. PMID- 11911244 TI - Interaction between bcl-2 and P53 in neoplastic progression of basal cell carcinoma of the head and neck. AB - BACKGROUND: Basal cell carcinoma (BCC) is the most frequent tumor of the human skin and generally shows a favourable clinical behaviour. However, a percentage of BCC grows aggressively, infiltrating contiguous structures, sometimes giving distant metastases. MATERIALS AND METHODS: Bcl-2 and p53 protein expression was studied immunohistochemically in 60 cases of BCC (30 non-aggressive, BCC1 and 30 aggressive cases, BCC2) of the head and neck region with a complete clinical follow-up. RESULTS: All the BCC1 showed distinct cytoplasmic staining for bcl-2. The intensity of staining ranged from intermediate to high, with only three cases showing low positivity. Among BCC2, none of the 30 cases showed positivity for bcl-2. Bcl-2 expression was directly correlated with the BCC1 sub-type and a favourable clinical follow-up (p<0.01). Among BCC1, 27 cases were found negative for p53 protein expression while 3 exhibited only a low immunoreactivity. Among BCC2, 11 out of 30 cases showed an intermediate immunoreactivity, and 18 out of 30 exhibited high positivity for p53 protein. The expression of p53 protein correlated inversely with cellular differentiation (p<0.01). CONCLUSION: From the analysis of these results it is reasonable to consider bcl-2 and p53 protein expression as useful discriminating prognostic factors in the evaluation of BCCs of the head and neck region. In fact, the finding of clones expressing bcl-2 in a case of BCC may be indicative of an "indolent" cellular neoplastic phenotype. In other words, bcl-2 could be used as a "clonal marker" of a still favourable clinical behaviour. Conversely, the partial or complete loss of bcl-2-bearing neoplastic clones during histological transformation, with the appearance of clones expressing p53 protein in a BCC could be considered a hallmark of transition from a low-to high-grade malignancy, characterized by the emergence of cellular clones with a more aggressive phenotype, responsible for worse clinical behaviour. PMID- 11911245 TI - Association between telomerase activity and basic fibroblast growth factor up regulation in retinoblastomas. AB - BACKGROUND: Telomerase is an enzyme associated with cellular immortality and malignancy in many cell types. On the other hand, growth factors such as basic fibroblast growth factor (bFGF) or platelet-derived growth factor (PDGF) promote tumor growth, whereas the association between telomerase and these growth factors remains unclear. MATERIALS AND METHODS: Telomerase activity and expression of bFGF and PDGF were assayed in 9 retinoblastoma tissues and 2 cell lines (WERI-Rb 1 and Y79). The association of telomerase activity with bFGF or PDGF was investigated. RESULTS: Telomerase activity was detected in three out of nine tissues and both cell lines. Two telomerase highly-positive tissues and WERI-Rb-1 and Y79 cells expressed bFGF. As for the expression of PDGF, only one retinoblastoma tissue with high telomerase was positive. To further determine whether telomerase activity and bFGF are closely associated, we inhibited each expression. Inhibition of telomerase in WERI-Rb-1 cells using the anti-sense treatment suppressed the expression of bFGF and subsequently induced apoptosis after 25 to 30 doublings. When bFGF expression was suppressed by the neutralizing antibody, telomerase activity was not affected nor was apoptosis detected. CONCLUSION: Telomerase may up-regulate the expression of bFGF and protect retinoblastoma cells from cell death, indicating, the possibility that inhibition of telomerase is a promising approach for the treatment of telomerase-positive tumors. PMID- 11911246 TI - Antiviral, cytotoxic and apoptotic activities of picolinic acid on human immunodeficiency virus-1 and human herpes simplex virus-2 infected cells. AB - The antiviral, cytotoxic and apoptotic effects of picolinic acid against human immunodeficiency virus-1 (HIV-1) and human herpes simplex virus-2 (HSV-2) infected cells were evaluated in cultured cells using in vitro assays. Picolinic acid was tested at several concentrations (from 0.15 to 3 mM) against one input dilution of the viruses to determine if the agent had any antiviral activity against HIV-1 or HSV-2. The results showed that picolinic acid at 1.5 mM (185 ug/mL) and 3 mM (369 ug/mL) was active against HIV-1 and HSV-2-infected cells, causing cytotoxicity which resulted in apoptosis and lack of viral replication. In parallel control experiments with non-infected cells, picolinic acid also caused cytotoxicity and apoptosis, which was more prominent at 3 mM than at 1.5 mM. Thus, infected cells appear to be slightly more sensitive to the cytotoxic and apoptotic effects of picolinic acid. The overlapping of the antiviral profile with the cytotoxic profile of picolinic acid indicates that, in vitro, the most likely sequence of events is that picolinic acid initially causes cytotoxicity which in turn results in apoptosis of cells infected with HIV-1 or HSV-2 and thus reduces the amount of viral replication. PMID- 11911247 TI - WP744, a novel anthracycline with enhanced proapoptotic and antileukemic activity. AB - BACKGROUND: MDR1 or MRP1 drug resistance mechanisms seriously limit the efficacy of anthracyclines such as doxorubicin, in the treatment of acute myeloid leukemia (AML). Our studies indicated that reducing basicity, increasing steric hindrance at C-4', and/or lipophilicity may help circumvent P-glycoprotein (P-gp)-mediated anthracycline efflux and thus increase drug retention in MDR-positive cells. From a series of 4'-substituted analogs, 4'-O-benzylated doxorubicin (WP744) was selected for a comparison with the classic anthracycline doxorubicin for their cytotoxic and pro-apoptotic properties. WP744 retains cytotoxic activity against P-gp and MRP-positive cells. METHODS AND RESULTS: In three AML cell lines (K562, KBM-3, and OCIM2) WP744 was markedly more potent (IC50 values of 0.18, <0.05, and <0.05 microg/ml, respectively) than doxorubicin (IC50 values of >0.5, 0.07, and 0.09 microg/ml, respectively). Likewise, WP744 inhibited the colony formation by AML-CFU cells from fresh bone marrow of three AML patients more strongly than doxorubicin. Cell growth inhibition by WP744 is accompanied by apoptosis induction as shown by TUNEL assay in OCIM2 cells. WP744-induced apoptosis appears to be mediated by caspase-3 as apoptotic changes were abrogated in the presence of the caspase 3 inhibitor Z-DEVD-FMK. Accordingly, caspase 3 activity was elevated in the lysates from drug-treated cells. WP744 induced also cleavage of apoptotic marker poly(ADP-ribose)polymerase (PARP). Finally, WP744 at 0.05 microM and greater was a potent inducer of apoptosis (by quantitative DNA fragmentation) in cultured human acute lymphoblastic leukemia (ALL) CEM cells, compared to 0.5 microM doxorubicin needed for a similar effect. CONCLUSION: The novel anthracycline WP744 was found to be an antileukemic agent with proapoptotic activity superior to that of doxorubicin. PMID- 11911248 TI - Antisense to integrin alpha v inhibits growth and induces apoptosis in medulloblastoma cells. AB - BACKGROUND: Integrin alpha v promotes brain microvessel endothelium survival, yet its role in brain tumor cells is unknown. MATERIALS AND METHODS: Alpha v synthesis in medulloblastoma cells Daoy, D341Med, and D283Med, was inhibited with antisense oligonucleotides (ASODN) to test the effect on growth and survivaL RESULTS: ASODN reduced alpha v surface expression 75% in a dose- and time dependent manner (2 microM, 72 hours). Alpha v-deficient cells grown with vitronectin demonstrated reduced cell spreading, G0-G1 growth arrest, decreased proliferation and increased apoptosis compared to controls or alpha v-deficient cells grown with collagen. Furthermore, insulin-like growth factor-I (IGF-I) suboptimally stimulated proliferation and survival of alpha v-deficient cells, suggesting that alpha v-IGF-I interactions potentiate medulloblastoma growth. Finally, treatment with alpha v-blocking antibody induced caspase-8 and caspase-3 expression, while apoptosis of alpha v-deficient cells was associated only with increased caspase-3. CONCLUSION: Alpha v integrin supports medulloblastoma growth by activating adhesion-dependent and -independent survival pathways and thus may serve as a novel therapeutic target in this tumor. PMID- 11911249 TI - Effect of daunomycin on expression of CD44H protein in HTB140 melanoma cells in vitro. AB - CD44H is a transmembrane protein involved in cell-cell and cell-matrix interactions. In melanoma cells CD44H influences motility and invasiveness. Daunomycin (daunorubicin) is an anthracycline antibiotic, capable to inducing apoptosis in many cell lines. The data presented in this report show that 12 hours treatment with clinically applied concentration of daunomycin led to apoptotic death of a portion of the investigated melanoma cell population. Surviving cells showed random distribution of CD44H and decreased expression of the protein on the cell surface, but no cell blebbing or changes in nuclei. Hypothetical mechanisms concerning a role of CD44H in induction of apoptosis are discussed. PMID- 11911250 TI - Identification of breast cancer metastasis-associated genes by chip technology. AB - In order to identify genes associated with metastasis of mammary carcinoma, we compared the transcriptional profile (Affymetrix chip technology) of two cell lines derived from primary mammary carcinoma, three cell lines derived from bone marrow micrometastasis, a cell line derived from a lymph node metastasis as well as a cell line derived from malignant ascites. We found that 11 genes (0.16%) were up-regulated in all five cell lines derived from metastasis and 32 genes (0.45%) were up-regulated in four of these cell lines. Sixteen genes (0.23%) were down-regulated in the five metastatic cell lines, while 24 genes (0.34%) were down-regulated in four of the metastatic cell lines. The usefulness of our system for the identification of genes associated with metastasis of mammary carcinoma is demonstrated by the identification of genes which have already been implicated in metastasis of mammary carcinoma. This suggests that further evaluation of identified de-regulated genes, which until now have not been seen in context with metastasis of mammary carcinoma, should be undertaken. PMID- 11911251 TI - Augmentation of apoptosis responses in p53-deficient L1210 cells by compounds directed at blocking NFkappaB activation. AB - A mouse leukemia L1210 cell line (Y8) selected for resistance to deoxyadenosine was found to be deficient in the expression of p53 mRNA and protein while maintaining the expression of WAF1/p21 mRNA and protein even under basal conditions. The Y8 cells were shown to be more sensitive to apoptosis induced by a variety of agents when compared to the parental wild-type (WT) L1210 cells. Roscovitine, an inhibitor of cdk 2 and cdk5, was one of the agents that caused increased apoptosis in the Y8 cells through a pathway that ultimately involved the activation of caspase-3 activity. In these studies, the effects of leflunomide and parthenolide (drugs reported to alter the activation of NFkappaB in a variety of cell types) were studied for their cell cycle and apoptotic effects in WT and Y8 cells as single agents and in combination with roscovitine. Leflunomide at IC50 concentrations had little effect on the cell cycle distribution of either the WT or Y8 cells while at higher concentrations caused a G0/G1 block in Y8 cells. Parthenolide, at IC50 concentrations, caused a G0/G1 cell cycle block in the WT and Y8 cells but at higher concentrations caused a G2/M block in the Y8 cells. The combinations of leflunomide and roscovitine or parthenolide and roscovitine did not alter, in a significant way the cell cycle distribution of the Y8 cells. However, in the presence of the combinations of leflunomide and roscovitine or parthenolide and roscovitine there were large increases in the fraction of Y8 cells undergoing early apoptosis without a corresponding increase in the necrotic fraction of cells. These data show that combinations of agents directed at different pathways or different steps of pathways involved in apoptosis can cause the cells to reach an apoptotic threshold that results in synergistic apoptosis. PMID- 11911252 TI - Involvement of cancer-activating genes on chromosomes 7 and 8 in esophageal (Barrett's) and gastric cardia adenocarcinoma. AB - The incidence of adenocarcinomas of the distal esophagus (Barrett's esophagus) and proximal stomach (gastric cardia) has increased rapidly over the past decades. In contrast to this dramatic increase, genetic knowledge is sparse. MATERIALS AND METHODS: We investigated genomic amplification on chromosomes 7 and 8 by comparative genomic hybridization (CGH) and protein expression of relevant oncogenes (EGFR, HGF, MET, CTSB, MYC) by immunohistochemistry (IHC) in 22 esophageal and 22 gastric cardia carcinomas. RESULTS: The CGH and IHC patterns were very similar for the two cancer locations. IHC showed positive immunostaining in 93% of the adenocarcinomas for at least one of the investigated genes, whereas CGH disclosed genomic gains on chromosome 7 and/or 8 in 80%. CONCLUSION: Cancer-activating genes on chromosomes 7 and 8 are frequently involved in gastro-esophageal junction adenocarcinomas. Moreover, the similarities in chromosomal changes and protein expression patterns strongly suggest that esophageal and gastric cardia adenocarcinomas have a shared etiology. This is in agreement with studies addressing gastroesophageal reflux disease and intestinal metaplasia at these locations. PMID- 11911253 TI - Matrix metalloproteinases (MMPs) in breast cancer cell lines of different tumorigenicity. AB - In the present study we investigated the presence, amount and activity of matrix metalloproteinases (MMPs)-1, -2, -3, -7, -8, -9, -10, -11 and -13 and TIMP-1 in three well-defined breast cancer cell lines with different biological behaviour; i.e. poorly-invasive MCF-7 cells, invasively growing MDA-MB-231 cells and invasive and highly-metastatic MDA-MB-435 cells. The parallel immunocytochemical determination of the degree of cellular differentiation, as monitored by the immunocytochemical expression of cytokeratins (CK), confirmed differences in the tumor cell differentiation. Thereby, MCF-7 cells expressed more glandular CKs than MDA-MB-231 cells, while MDA-MB-435 cells were only labelled by pancytokeratin markers, but neither by glandular nor by squamous epithelial CKs. Conditioned media were analyzed for the presence of MMPs and TIMP-1 using Western blot with specific polyclonal antibodies and for gelatinolytic and caseinolytic activity by zymography. In addition, the cellular pool of several MMPs was investigated by immunocytochemistry. An enhanced cytoplasmatic staining for MMP-3 and -9, MMP-1, -10 and -11 was seen in the highly metastatic cells at almost equal levels, while MMP-2 revealed only a minor intracellular staining in all three cell lines. Western blots of conditioned media showed enhanced amounts of MMP-1, -3, -7, -10 and -11 in media of the two metastatic cell lines. Casein zymography correlated with the results of the MMP-1 Western blots. By means of gelatin zymography, MMP-2 and -9 were detectable in cell culture supematants of all the three cell lines, while gelatinolytic activity was elevated in the media of the more malignant MDA-MB-435 cells. Separate addition of EDTA or Pefa bloc SL partially inhibited the gelatinoltic activity indicating the presence of metallo- and serine proteinases, respectively; combined application of both inhibitors resulted in a complete suppression of activity. We provide evidence that the deviation expression in secretion of various MMPs in breast cancer cell lines of different tumorigenicity correlates with the biological behaviour of these cells, ie. the more malignant cells synthesize more MMPs than the less malignant ones. In addition, the secretion of MMP-1, -3, -7, -10 and -11 was enhanced in the malignant MDA-MB-231 and -435 cells when compared to the corresponding intracellular pool. This analysis confirms previous results obtained in a keratinocyte tumor cell model and provides evidence for a more general biological association between MMP-expression and tumor cell growth. PMID- 11911254 TI - A quantitative angiogenesis model for efficacy testing of chemopreventive agents. AB - One of the approaches in chemoprevention to prevent or delay the progression of precancerous lesions, is to apply chemopreventive agents that can potentially block angiogenesis. A quantitative in vivo angiogenesis inhibition assay was developed to test the efficacy of twelve chemopreventive agents that represent different chemical classes and multiple biological activities, using the chick chorioallantoic membrane (CAM) model and an oncogene-transfected angiogenic cell line (6 Ti ras/SV myc # 4). These tumorigenic cells held by a primary agarose pellet, were placed alone or with a secondary pellet incorporating five concentrations of the test agent, on an exposed CAM of 7-day-old chick embryo for 72 hours in a humidified chamber at 35 degrees C. The cell-induced angiogenic blood vessels, including the microvessels radiating from the cell pellet focal area, were scored using a computerized custom image analysis system. The results show that nonsteroidal antiinflammatory drugs (NSAIDS); aspirin, sulindac, sulindac sulfide and sulindac sulfone, were effective inhibitors of cell-induced angiogenesis (23-66%). Aspirin displayed a dose-dependent response with the highest inhibition at 300 microM and an EC50 (the effective molar concentration that inhibits angiogenesis by 50%) of 26 microM. Sulindac sulfone was more effective than sulindac with an EC50 of 5 microM versus 85 microM. However, sulindac sulfide showed an intermediate response with an EC50 of 41 microM. The retinoids; all-trans-retinoic acid (ATRA), 9-cis-retinoic acid (9-cis-RA), and 13 cis-retinoic acid (13-cis-RA) were also highly effective inhibitors of cell mediated CAM-angiogenesis. 13-cis-RA with an EC50 of 3.6 nM, has been the most efficacious test agent. > 400-fold more effective than 9-cis-RA (1.5 microM). ATRA exhibited an intermediate response between 9-cis-RA and 13-cis-RA with an EC50 of 0.3 microM, and was 100-fold more efficacious than 9-cis-RA. However, the synthetic retinoid, N-(4-hydroxyphenyl) retinamide (4-HPR), was not an effective inhibitor of CAM angiogenesis. Thalidomide, a compound with multiple biological activities, exhibited dose-dependent inhibition ranging from 10-1000 microM with an EC50 of 19 microM. Other agents that exhibited dose-dependent inhibition included Bowman-Birk inhibitor (BBI), EC50: 10 microg/ml, tamoxifen, EC50, 0.05 microM and difluoromethyl omithine (DFMO), with an EC50 of 13 microM. These results suggest that tumor-associated angiogenesis can be modulated by non-toxic concentrations of chemopreventive agents representing multiple biological activities and multiple targets. PMID- 11911255 TI - Chemotherapeutic evaluation of 4-hydroxybenzylretinone (4-HBR), a nonhydrolyzable C-linked analog of N-(4-hydroxyphenyl) retinamide (4-HPR) against mammary carcinogenesis. AB - The antitumor effects of N-(4-hydroxyphenyl)retinamide (4-HPR), and its stable C linked analog, 4-hydroxybenzylretinone (4-HBR) on the regression of established 7,12-dimethylbenz(a)anthracene(DMBA)-induced rat mammary tumors were compared. 4 HBR is a stable and nonhydroyzable derivative which cannot be converted in vivo to retinoic acid (RA). The results indicate that 4-HBR decreased mammary tumor volumes to the same extent as equimolar concentration (2 mmol/kg diet) of 4-HPR ( 45% for 4-HBR vs. -42% for 4-HPR, p<0.01). Both 4-HPR and 4-HBR bind very poorly to nuclear retinoid receptors RARs and RXRs. The similarity of physicochemical properties of 4-HPR and 4-HBR as well as their equal antitumor potency suggests that 4-HPR like 4-HBR, is acting directly rather than through hydrolysis to free RA. Treatment with 4-HPR caused an almost 65% decrease in serum retinol levels. These results suggest that 4-HBR may have a significant chemotherapeutic advantage over 4-HPR, as the nonhydrolyzable analog may not cause night blindness which occurs as a significant side effect of 4-HPR usage. PMID- 11911256 TI - Effect of a matrix metalloproteinase inhibitor (ONO-4817) on lung metastasis of murine renal cell carcinoma. AB - We examined the anti-metastatic effect of a newly developed inhibitor of synthetic matrix metalloproteinase (MMP), ONO-4817, on experimental pulmonary metastasis of murine renal cell carcinoma (Renca) cells and on tumor cell invasion, through reconstituted basement membrane (Matrigel) in vitro using the same cells. Oral administration of ONO-4817 (50-200 mg/kg/day) to Renca-bearing mice resulted in a dose-dependent inhibition of lung metastasis without a loss of body weight. ONO-4817 at the high dose of 200 mg/kg showed a tendency to prolong the survival of the mice. We also found that oral administration of ONO-4817 significantly inhibited the angiogenic response (number of vessels oriented towards the tumor mass) and the growth of tumors inoculated i.d. in syngeneic mice. In addition, ONO-4817, at non-cytotoxic concentrations of less than 10 microM, caused a marked inhibition of the invasion of Renca cells as compared to the vehicle control. Gelatin zymography revealed that ONO-4817 inhibited the enzymatic activity of MMP-2 produced by Renca cells in a concentration-dependent manner. In conclusion, ONO-4817 effectively inhibited lung metastasis of Renca cells through its anti-invasive and anti-angiogenic properties. These results suggest that use of the MMP inhibitor (MMPI) ONO-481 7 may provide a therapeutic basis for preventing lung recurrence and metastasis of renal cell carcinoma. PMID- 11911257 TI - The frequency of micronuclei in lung cancer cell lines and their correlation to intrinsic radiation sensitivity. AB - BACKGROUND: Micronuclei arise from chromosomal, acentric, fragments or whole chromosomes that are not incorporated into the daughter nuclei at mitosis. The micronuclei assay is technically simple and requires only one mitosis in order to obtain information concerning the amount of micronuclei. This study was performed to investigate whether the formation of micronuclei could be used as a marker for intrinsic radiation sensitivity in lung cancer patients. MATERIALS AND METHODS: Two human lung cancer cell lines, a non-small cell lung cancer (NSCLC) cell line (U-1810) and a small cell lung cancer (SCLC) cell line (U-1906L), were used. Radiosensitivity data on the survival fraction at 2 Gy were obtained from the clonogenic assay. Radiation was delivered as one-fraction doses: 0, 1, 2, 4, 10 and 20 Gy. After irradiation, the cells were incubated for 0, 24, 48, 60, 72 and 96 hours before fixation and staining. RESULTS: The frequency of micronuclei in U 1906L was clearly elevated after 96 hours in the 20 Gy fraction. The frequency of micronuclei reached 5.5%. For U-1810 the micronuclei had a peak clearly different than the other settings after 48 hours in the 10 Gy fraction. The frequency of micronuclei was 1.2%. CONCLUSION: Counting micronuclei is not sensitive enough for estimation of radiosensitivity in clinical doses. However, our results demonstrated a distinct difference between NSCLC and SCLC cell lines at higher doses. This difference might be due to different repair fidelity, so future studies with this assay should aim to investigate this hypothesis. PMID- 11911258 TI - Is magnetic resonance imaging texture analysis a useful tool for cell therapy in vivo monitoring? AB - Assessment of anti-tumor treatment efficiency is usually done by measuring tumor size. Treatment may however induce changes in the tumor other than tumor size. Magnetic Resonance Imaging Texture Analysis (MRI-TA) is presently used to follow activated lymphocyte cell therapy. We used a 7T microimager to acquire high resolution MR images of an experimental liver metastasis from colon carcinoma in rats treated (n = 4) or not (n = 3) with a cell therapy product. MRI-TA was then performed with Linear Discriminant Analysis and showed: i) a significant variation of tumor texture with tumor growth and ii) a significant modification in the texture of tumors treated with activated lymphocytes compared with untreated tumors. T2-weighted images or volume calculation did not evidence any difference. MRI-TA appears as a promising method for early detection and follow up of response to cell therapy. PMID- 11911259 TI - Gene expression profiles in human BPH: utilization of laser-capture microdissection and quantitative real-time PCR. AB - Human benign prostatic hyperplasia (BPH) nodules consist of epithelium and stroma that have different properties functionally as well as morphologically. To investigate the molecular profiles of the prostate gland, separate examination of these components are necessary. Laser-capture microdissection (LCM) is a newly developed device which enables one to dissect interesting parts of tissues under the microscope. In the current work we studied the gene expression profiles of prostatic epithelium and stroma using LCM. Androgen receptor (AR), estrogen receptor alpha and beta (ER alpha and ER beta), progesterone receptor (PR) and 5 alpha reductase type I and type II (5alphaR I and 5alphaR II) gene expressions were studied by RT-PCR. All of these genes were expressed both in the epithelium and stroma. Furthermore, AR transcript was quantified by quantitative real-time PCR. AR transcript ranged from 302 to 1440 copies per 10(5) GAPDH copies and from 257 to 3223 copies per 10(5) GAPDH copies in the epithelium and stroma, respectively. Thus, LCM and quantitative real-time PCR are powerful tools for molecular analysis of heterogeneous tissues including prostate gland. PMID- 11911260 TI - Evaluation of a selective prostaglandin E receptor EP1 antagonist for potential properties in colon carcinogenesis. AB - BACKGROUND: Cyclooxygenases (COXs) and prostanoids play pivotal roles in colon carcinogenesis. This study was designed to determine the chemopreventive effects of ONO-8711, a selective prostaglandin E receptor EP1 antagonist, on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) in male F344 rats and to compare its potential with that of nimesulide, a well documented selective COX-2 inhibitor. MATERIALS AND METHODS: Five-week-old male F344 rats received s.c. injections of AOM (15 mg/kg body weight) or the saline vehicle once weekly for two weeks and were fed the control diet (AIN-76A) or the experimental diets containing 400 or 800 ppm of ONO-8711 or 400 ppm nimesulide for 5 weeks. RESULTS: Administration of ONO-8711 at 800 ppm significantly reduced the total number of ACF/colon and 5-bromodeoxyuridine (BrdUrd) labeling index as compared to the control diet group (by 31% and 66%, respectively). As expected, dietary administration of nimesulide also suppressed the development of ACF and BrdUrd labeling index in the colon, by about 39% and 54%, respectively. CONCLUSION: Our finding that ONO-8711 significantly suppresses colonic ACF formation and cell proliferation strengthens the hypothesis that the selective prostaglandin E receptor EP1 antagonists possesses chemopreventive activity against colon cancer development. PMID- 11911261 TI - Progesterone induces apoptosis in malignant mesothelioma cells. AB - Progesterone has been used in the treatment of patients with recurrent or metastatic progesterone receptor-positive endometrial carcinoma and breast cancer. In vitro study using a breast cancer cell line, T47D, demonstrated an increase in p53 gene expression and induction of apoptosis by the administration of progesterone. Therefore, we investigated the effect of progesterone administration on the proliferation and apoptosis in a mesothelioma cell line, 211H. The expression of the progesterone receptor gene was detected in this cell line by a nested RT-PCR method. The proliferation of the cell line was suppressed after a 10-day incubation with 30 microM progesterone. In progesterone-treated 211H cells, apoptotic cells were detected by TUNEL assay and nuclear DNA fragmentation analysis. These results clearly demonstrated that progesterone administration suppressed the cell proliferation and induced apoptosis in malignant mesothelioma cells. PMID- 11911262 TI - Cytotoxic activity of gallic acid against liver metastasis of mastocytoma cells P 815. AB - Gallic acid (3,4,5-trihydroxy benzoic acid), a naturally occurring plant phenol, inhibited the proliferation of metastatic tumor cells, such as P815 murine mastocytoma, B16 murine melanoma and L5178 murine lymphoma cells at IC50s of 6.5, 8.0 and 3.6 microg/ml, respectively. P815 mastocytoma cells are known to metastasize specifically to the liver. When DBA/2 mice, injected intravenously with P815 cells, were treated with gallic acid at a concentration of 50 mg/kg, the number of nodules in the liver and serum glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT), which had increased as liver metastasis progressed, decreased. However, gallic acid itself did not show a liver protective effect though the life span of DBA/2 mice was extended by gallic acid treatment. These results suggest that gallic acid is able to inhibit liver metastasis, by killing P815 cells metastasized to the liver. PMID- 11911263 TI - Variation of HMGB1 expression in breast cancer. AB - The amount of steroid hormone receptor proteins does not always correlate with the response of breast cancers to endocrine therapy. This may partly be due to the fact that binding of the estrogen receptor (ER) to estrogen responsive elements (ERE) of its target genes is mediated by additional cellular proteins. One of these is the high mobility group protein HMGB1, known to interact with ER thus dramatically increasing its binding to ERE. This is the first report analysing the expression patterns of HMGB1 in breast cancer cells. Northern blot analyses of the 1.4 kb and the 2.4 kb transcripts of HMGB1 in 13 breast cancer samples revealed a strong intertumoural variation by a factor of 8.5 and 14.5, respectively. This variation may contribute to the different response, of estrogen receptor-positive breast tumours to endocrine therapy, making HMGB1 a marker of considerable clinical interest. PMID- 11911264 TI - Therapeutic electromagnetic field effects on angiogenesis and tumor growth. AB - BACKGROUND: A new approach to cancer therapy based on the application of therapeutic electromagnetic fields (TEMF) has been developed by EMF Therapeutics, Inc., Chattanooga, TN, USA. This study was designed to assess the effect of TEMF on tumor vascularization and growth of murine 16/C mammary adenocarcinoma cells in C3H/HeJ mice. MATERIALS AND METHODS: Implanted tumors were allowed to grow for seven days until the tumor volume reached 100 mm3 before treatment was started. Mice (20 per control, 10 per EMF exposed group) received treatment (10 minutes per day with 0, 10 mT, 15 mT or 20 mT) with a 120 pulses per second pulsating magnetic field. Tumor growth was assessed throughout the treatment period. The extent of tumor vascularization was evaluated by immunohistochemical staining for CD31. RESULTS: Exposure to TEMF significantly reduced tumor growth, significantly reduced the percentage of area stained for CD31 indicating a reduction in the extent of vascularization and there was a concomitant increase in the extent of tumor necrosis. CONCLUSION: A novel TEMF treatment safely reduced growth and vascularization of implanted breast cancers in mice. IMPLICATION: TEMF may prove a useful adjuvant to increase the therapeutic index of conventional cancer therapy. PMID- 11911265 TI - Antiangiogenic and antitumour effects in vivo of genistein applied alone or combined with cyclophosphamide. AB - The antitumour and antiangiogenic effects in vivo of genistein, applied alone or in combined therapy with cyclophosphamide, in the Lewis lung carcinoma (LL2) and B16 melanoma mouse tumour models, were analysed. Our own new method, allowing quantification of the volume of blood present in tumour tissue, enabled estimation of the degree of vascularization. Tumour cells entrapped in alginate beads were injected subcutaneously into mice. The quantification of alginate implant vascularization was performed with 125I-labeled mouse albumin injected intravenously. In mice bearing transplantable Lewis lung cancer the additive antiangiogenic, but not cytostatic, effect of genistein combined with cyclophosphamide (CY) was observed, since the treatment with genistein alone reduced tumour blood supply in 35% (tumour weight in 36%), with CY in 38% (tumour weight in 70%) and with both compounds in 61% (tumour weight in 75%). In the B16 melanoma model the respective values were: 60 and 44% for genistein, 83 and 79% for CY and 76 and 74% for combined treatment. These results indicate a higher antiangiogenic rather than cytostatic effect of genistein in both mouse tumour models applied. PMID- 11911266 TI - In vitro antitumor activity of TAS-103 against freshly-isolated human colorectal cancer. AB - DNA topoisomerases (Topo) are enzymes that relieve the secondary twist on the DNA strand in the process of DNA synthesis and transcription; therefore they are unique targeting molecules for the chemotherapy of colorectal cancer. TAS-103 (6 [[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-in-deno [2,1-c]quinolin-7-one dihydrochloride, MW; 406.31), a novel quinoline derivative, has recently been established as a Topo I and Topo II inhibitor. The aim of the present study was to investigate the antitumor activity of TAS-103 by the MTT assay in human highly purified and freshly-isolated colorectal cancer cells. To our knowledge, this is the first data concerning the antitumor activity of TAS-103 in highly-purified and isolated human colorectal cancer cells. TAS-103 showed the strongest antitumor activity among the conventional anticancer agents for colorectal cancer (p<0.05). The combination with CDDP augmented the antitumor activity of TAS-103 (p<0.05), indicating that CDDP is one of the most potent candidates to be used in combination with TAS-103. To predict the clinical effect of TAS-103, the expressions of Topo I and Topo II were measured by quantitative PCR. However, a correlation between the expression of Topo and the antitumor activity of TAS-103 was not established. In conclusion, according to this data, TAS-103 may be useful in the chemotherapy of colorectal cancer. PMID- 11911267 TI - Strong antioxidant activity of ellagic acid in mammalian cells in vitro revealed by the comet assay. AB - Oxidative stress due to oxygen and various radical species is associated with the induction of DNA single- and double-strand breaks and is considered to be a first step in several human degenerative diseases, cancer and ageing. Naturally occurring antioxidants are being extensively analysed for their ability to protect DNA against such injury. We studied three naturally occuring compounds, Ascorbic Acid, Melatonin and Ellagic acid, for their ability to modulate DNA damage produced by two strong radical oxygen inducers (H2O2 and Bleomycin) in cultured CHO cells. The alkaline Comet assay was used to measure DNA damage and a cytofluorimetric analysis was performed to reveal the intracellular oxidative species. The data showed a marked reduction of H2O2- and Bleomycin-induced DNA damage exerted by Ellagic Acid. On the contrary Ascorbic acid and Melatonin appeared to induce a slight increase in DNA damage per se. In combined treatments, they caused a slight reduction of H2O2-induced damage, but they did not efficiently modulate the Bleomycin-induced one. The Dichlorofluorescein diacetate (DCFH-DA) cytofluorimetric test confirmed the strong scavenging action exerted by Ellagic Acid. PMID- 11911268 TI - Modulation of CD99/MIC2 expression of human AHTO-7 osteoblasts by carcinoma cell line-conditioned media. AB - BACKGROUND: Prostate cancer metastases induce a predominantly osteoblastic response in bone tissue, resulting in new bone formation and associated morbidity; however, the mechanisms of these tumor-host responses are not fully understood. MATERIALS AND METHODS: Supernatants of prostate (PC3, DU145, LNCaP), breast (BT20, ZR-75-1), colon (SW620, Colo 320DM), pancreatic (ASPC1, Capan-1), renal cell (ACHN) and hepatoma (HepG2) cell lines were tested for their capacity to modulate proliferation, activity of alkaline phosphatase (ALP) and CD99/MIC2 expression in AHTO-7 (large T antigen transfected human trabecular osteoblasts) cells in vitro. RESULTS: Osteoblastic stimulation is not restricted to prostate cancer derived conditioned media CM and high activity is found in CM from Capan 1, HepG2 and ACHN lines. Furthermore these CM down-regulate the expression of the CD99/MIC2 antigen in comparison to medium by AHTO-7 cells as detected by HBA-71 immunofluorescence, with the exception of prostate cancer-derived CM. Induction of the differentiation marker ALP was detected in response to CM derived from Capan-1, BT-20 and Colo320DM. Stimulation of the proliferation of AHTO-7 cells (105-138% of control), induction of ALP (1.17-5.29-fold) and down-regulation of CD99 (13.6-57.5%) exhibited no correlation. CM derived from PC3 and LNCaP metastatic prostate cancer cell lines specifically resulted in the retention/stimulation of the expression of CD99/MIC2 in AHTO-7 cells in contrast to all other cell lines tested. CONCLUSION: The CD99/MIC2 antigen, which is expressed on human osteoblasts and osteosarcoma cells, seems to constitute a new and independent response marker of osteoblasts in the triggering of osteoblastic reaction by prostate cancer cells. PMID- 11911269 TI - Differentiation of the P-gp and MRP1 multidrug resistance systems by mobile lipid 1H-NMR spectroscopy and phosphatidylserine externalization. AB - We have previously demonstrated that proton NMR spectra of fatty acid chains in erythroleukemia K562 wild-type cells and their MDR1 counterparts show variations related to the phenotype over-expressing the P-glycoprotein (P-gp). Human lung cancer cells whose multidrug resistance (MDR) counterparts over-express the multidrug resistance-associated protein MRP1 have not yet been studied by NMR. Both P-gp and MRP1 belong to the same ATP-binding cassette transporter superfamily. A comparison of NMR spectra from both these multidrug-resistance phenotypes showed that the results previously obtained on the MDR1 family are not valid for MRP1. Furthermore, flow cytofluorimetry studies with external phosphatidylserine labelling showed that P-gp and MRP1 overexpressions have strong but differentiated effects on cell lipid pools. PMID- 11911270 TI - Analysis of EBV latency by EBER in situ hybridization in nasopharyngeal carcinoma Spanish patients. AB - BACKGROUND: The aim of this study was to analyse the Epstein-Barr virus (EBV) latency by detecting the EBV-associated latent small nuclear RNAs (EBER), in a group of biopsies from Spanish patients with diagnosed nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: NPC paraffin samples with the presence of EBV demonstrated by non-isotopic in situ hybridization (NISH) and nested-PCR, were analysed for EBV latency using EBER in situ hybridization (EBER-ISH). RESULTS: We detected EBER in 83.3% of samples (10 out of 12 cases), demonstrating the relationships between EBV genome presence with the latent viral infection. We correlated these results of EBV-DNA and -RNA presence with the immunoexpression of latent membrane protein-1 (LMP-1), a viral oncogenic protein (8 out of 12 cases or 66.6%). CONCLUSION: These results indicate that all the types of NPC are variants of an EBV-associated malignancy and that viral latency is a critical phenomenon in the development of this neoplasia. PMID- 11911271 TI - Radiation responsiveness of human lung cancer cell lines measured with a short term semiautomatic assay. AB - BACKGROUND: Fluorometric microculture cytotoxicity assay (FMCA) is a short-term semi-automatic method, based on dye-inclusion of surviving cells. The assay was developed for investigations of drug resistance on tumour cells from biopsy material. In the present study, this short-term assay was evaluated, regarding usefulness in determining radio-sensitivity. MATERIALS AND METHODS: Eight human lung cancer cell lines were used. There were five small cell lung cancer (SCLC and three non-small cell lung cancer (NSCLC cell lines. Results were compared with the corresponding data derived from the clonogenic assay and/or the extrapolation method. RESULTS: The surviving fraction (SF) after 2, 5 and 10 Gy compared with data from the clonogenic assay were not in accordance for 5 of the 8 cell lines. The FMCA assay overestimated SF- values for the SCLC cell lines. CONCLUSION: The FMCA assay is not useful as a quick screening method for the radioresponsiveness in vitro of human tumour cell lines. PMID- 11911272 TI - Induction of apoptotic pathway in the vestibule of cisplatin (CDDP)-treated guinea pigs. AB - BACKGROUND: During the process of apoptosis, double-stranded DNA is broken into single-stranded DNA by the action of caspases and caspase-activated deoxyribonuclease. We immunohistochemically examined the apoptotic changes induced by cisplatin in the vestibule of guinea pigs. MATERIALS AND METHODS: Cisplatin (10 mg/kg b.w.) was intraperitoneally injected into guinea pigs and, 3 days after the injection, the animals were sacrificed by intracardiac perfusion of fixative. The temporal bones were then removed and immunohistochemically stained for caspase-activated deoxyribonuclease or caspase 3. RESULTS: Both caspase-activated deoxyribonuclease and caspase 3 were observed in the dark cell area, transitional area and the sensory epithelium. CONCLUSION: These findings suggest that apoptosis is involved in the vestibular dysfunction of the CDDP treated patients. PMID- 11911273 TI - Carboplatin AUC and gamma-glutamylcysteine synthetase gene expression in peripheral mononuclear cells of lung cancer patients. AB - BACKGROUND: To investigate the association between glutathione-related enzymes and carboplatin (CBDCA) dose, we examined gene expression levels for both subunits of gamma-glutamylcysteine synthetase (heavy; gamma-GCSh, light; gamma GCS1) in peripheral mononuclear cells (PMN) of lung cancer patients before and after CBDCA administration. MATERIALS AND METHODS: PMN and plasma samples were obtained from 10 advanced non-small lung cancer patients before and after CBDCA administration. We analyzed the gene expression levels by reverse transcription polymerase chain reaction. RESULTS: Gamma-GCSh expression levels in PMN increased within 24 hours after CBDCA administration, whereas gamma-GCS1 expression levels did not. However, the actual area under the concentration curve (AUC) of CBDCA did not correlate with gamma-GCSh expression at 24 hours or the increased ratio of gamma-GCSh expression in PMN. CONCLUSION: Expression of gamma-GCSh is induced by CBDCA, however, CBDCA AUC is not a determinant for the increased expression levels of gamma-GCSh in PMN. PMID- 11911275 TI - Potential of the proteasomal inhibitor MG-132 as an anticancer agent, alone and in combination. AB - Proteasomal activity is required for normal cellular functions including cell division, where entry and exit from mitosis is strictly regulated by cyclins and cyclin-dependent kinases which are among the important substrates of the proteasomal degradative machinery. Inhibitors of proteasomal activity have been shown to be effective inducers of apoptosis in tumor cells and may be useful as anticancer agents, either alone or in combination with other drugs. We have examined the effect of MG-132, a dipeptide proteasomal inhibitor, on various human cancer cell lines. We have also examined the effect of MG-132 on normal CD34+ enriched primary human peripheral blood stem cells. Our results indicate that MG-312 is a potent anticancer agent with cytotoxic effects on a variety of human cancer cell lines irrespective of their p53 status. MG-132 was found to be more effective in combination with drugs such as doxorubicin and etoposide that act in the S/G2-phase of the cell cycle via a mechanism that involves stabilization of cyclin B1 and increased expression of Bax. Further, MG-132 inhibits CFU-GM colony formation of the CD34+ enriched PBSC population and this inhibition correlates with release of cyt C into the cytosol. PMID- 11911274 TI - Long-term effects of 1-nitropyrene on oncogene and tumor suppressor gene expression. AB - Late changes in the expression of oncogenes and tumor suppressor genes following carcinogenic exposure were examined in lung, liver and kidney. 1-Nitropyrene (1 NP), which is a high-risk exposure factor in urban and industrial zones, was used as a carcinogenic agent. c-myc, Ha-ras and p53 gene expression was investigated after administration of a single dose of 1-NP to sensitive CBA/Ca mice in lung, liver and kidney for one year. One week after a single dose 1-NP administration, the expression of p53 was elevated in the liver, but, decreased in the lung and kidney. There was no increase in the expression of c-myc or Ha-ras genes at that time. One month after the administration of the 1-NP, the expression of p53 was increased in the kidney while the expression of Ha-ras and p53 was elevated in the liver. There was no significant difference in gene expression between the treated and control animal groups at any of the investigated periods except for the above-mentioned organs and at the end point of the investigation. According to the literature, 1-NP and its metabolites remain at high concentrations in the kidney, liver and lung. The concentration of the carcinogenic agent and the expression of the studied genes did not seem to correlate with each other in this experiment. PMID- 11911276 TI - Chalcones: structural requirements for antioxidant, estrogenic and antiproliferative activities. AB - Flavonoids are largely studied for their biological properties and particularly for their scavenging and antioxidant activities. In the present study, we first evaluated the antioxidant and the estrogenic actions of chalcones, then we tested their effects on MCF-7 cell proliferation. Chalcones are unique in the flavonoids family in lacking a heterocyclic C ring. We tested substituted chalcones with different numbers and different positions of the hydroxy groups: 2' hydroxychalcone, 4'-hydroxychalcone, 4-hydroxychalcone, 2',4-dihydroxychalcone, isoliquiritigenin, 2',4'-dihydroxychalcone, phloretin and naringenin chalcone. For the antioxidant tests we established the importance of the alpha-beta double bond and the 6'-hydroxy group. The establishment of the structure-activity relationship for the estrogenic properties showed a correlation between the antioxidant and the estrogenic properties. The importance of conformation and hydroxy group positions observed for chalcones, having antioxidant and estrogenic properties, was also observed on MCF-7 cell growth with the same structure activity relationship. The role of electron and hydrogen transfer in the correlation between these three biological activities was discussed. PMID- 11911277 TI - Antileukemic activity of synthetic daunomycinone derivatives bearing modifications in the glycosidic moiety. AB - The antileukemic activities of the daunomycinone glycosides synthesized in our laboratories (compounds 4 and 7, code names S12 and S13, respectively) were characterized in L1210 cells in vitro. S13 inhibits tumor cell proliferation and viability at day 4 (IC50: 150-200 nM) more effectively than S12 (IC50: 250-450 nM), suggesting that the 4'-trifluoracetamido substitution of the glycosidic moiety of these 3'-halo daunonycinone derivatives has greater antitumor potential than the 4'-azido substitution. Since S12 and S13 do not increase but rather decrease the mitotic index of L1210 cells at 24 hours, they are not antitubulin drugs but might arrest the early stages of cell cycle progression. Pretreatments for 1.5-3 hours with S12 and S13 are sufficient to partially inhibit the rates of DNA and RNA syntheses (IC50: 4-10 microM) determined over 30- to 60-minute periods of pulse-labeling in L 1210 cells in vitro, but these daunomycinone glycosides alter neither the cellular transport of purine and pyrimidine nucleosides nor the rate of protein synthesis. After 24 hours, the concentration dependent induction of DNA cleavage by S13 reaches a plateau at 10 microM but the weaker S12 requires 48 hours to maximally stimulate DNA cleavage like S13. The mechanism by which S13 induces DNA fragmentation is inhibited by actinomycin D, cycloheximide, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethyl ketone, N-tosyl-L-phenylalanine chloromethyl ketone and ZnSO4, suggesting that S13 triggers apoptosis by caspase and endonuclease activation. Since microM concentrations of S12 and S13 are cytostatic and cytotoxic, but do not sufficiently inhibit RNA and protein syntheses to block their own ability to sustain the active process of apoptosis and DNA fragmentation, such 3'-halo daunomycinone glycosides might be valuable to develop new means of polychemotherapy. PMID- 11911278 TI - Up-regulation of telomerase activity in Herpesvirus saimiri immortalized human T lymphocytes. AB - Human T-lymphocytes can be transformed to unlimited growth by Herpesvirus saimiri (HVS). We studied the telomerase activity of a recently established HVS immortalized human CD4 T cell clone in comparison to peripheral blood lymphocytes (PBL) and unstimulated or phytohemagglutinin (PHA)-stimulated CD4 T-cells by a Telomeric Repeat Amplification-Protocol (TRAP) -Assay. Telomerase activity in PHA stimulated CD4 T-cells was seven-fold and in HVS-infected CD4 T-cells 14-fold higher than in untreated CD4 T-cells. The HVS immortalized T-cell clone provides a useful tool for studying the regulation of telomerase activity during carcinogenesis and for testing of telomerase-inhibitory drugs. PMID- 11911279 TI - Remodeling of vimentin cytoskeleton correlates with enhanced motility of promyelocytic leukemia cells during differentiation induced by retinoic acid. AB - The intermediate filament (IFs) cytoskeleton is one of the major determinants for the mechanical properties of cytoplasm. Vimentin is the major IFs protein in peripheral blood neutrophils. We investigated its expression and function during neutrophil differentiation using the promyelocytic leukemia cell line NB4. The differentiation of NB4 cells along the neutrophil lineage and the monocytic pathway was induced by all-trans retinoic acid (ATRA) and phorbol esters (PMA), respectively. We demonstrated a down-regulation of vimentin after ATRA treatment of NB4 cells by immunoblotting and immunofluorescence. The architecture of the vimentin cytoskeleton in differentiated NB4 cells resembled that observed in mature neutrophils. In contrast, we showed a slight increase of vimentin content in phorbol ester (PMA)-treated NB4 cells. The structural features of the vimentin cytoskeleton obtained by image analysis showed significant differences in network density and directionality between ATRA-treated NB4 cells and controls. The functional consequence of the cytoskeletal remodeling for the mechanical properties of NB4 cells was assessed in migration assays. After ATRA treatment, we found a 4-fold increased migration of NB4 cells across transwell membranes with a 8 microm pore size without any cell size modification. No significant differences between PMA-treated NB4 cells and control cells could be observed using similar tests. These results indicate that both vimentin expression and network architecture are tightly controlled during neutrophil differentiation to regulate the mechanical properties of these cells. PMID- 11911280 TI - Anticancer activity of rViscumin (recombinant mistletoe lectin) in tumor colonization models with immunocompetent mice. AB - The antitumoral and immunostimulating properties of rViscumin (recombinant mistletoe lectin) were investigated in two mouse tumor models. After intravenous inoculation with RAW-117-P or L-1 sarcoma cells in Balb/c mice, rViscumin was given s.c. at non-toxic doses ranging from 0.3 to 150 ng rViscumin/kg. One set of experiments was performed to investigate the survival of rViscumin-treated animals. Another set was carried out to analyze the effect of rViscumin treatment on the number of tumor colonies in infiltrated lungs (RAW-117P) or liver (L-1) and the activation of immune cell subsets, respectively. An overall prolonged survival time after treatment with rViscumin and a reduction in the number of tumor colonies after administration of certain rViscumin doses was observed. Immunophenotyping of the peripheral leukocytes of treated mice revealed increased numbers of T-lymphocytes, pan-NK cells and activated monocytes. The results indicate that rViscumin has antineoplastic properties and might therefore be a promising candidate in cancer therapy. PMID- 11911281 TI - Interferon-gamma inhibits growth and migration of A172 human glioblastoma cells. AB - Malignant gliomas are highly proliferative and invasive tumors with poor prognosis. We investigated the influence of Interferon-gamma (IFN-gamma) on the human malignant glioma cell line A172, measuring cell viability (MTT-test), proliferation (3H-thymidine-uptake), cell death (FACS) adhesion to hyaluronic acid (HA, adhesion-assay) and migration (Boyden-chamber). IFN-gamma significantly decreased cell viability and proliferation. Measured by FACS, an up-regulation of CD95 expression has been shown in combination with an increased rate of cell death, first seen after 96 hours IFN-gamma treatment. Adhesion to HA was decreased after pre-treatment with IFN-gamma. This was not mediated by down regulation of the main HA-receptor CD44, since IFN-gamma did not change CD44 expression. IFN-gamma-treated cells showed a significantly diminished migration rate through a native or HA-coated 8-microm polycarbonate membrane. To summarise, IFN-gamma influences both the main characteristics of malignancy: it decreases cell proliferation and induces cell death, further it diminishes migration of A172 human glioblastoma cells. PMID- 11911282 TI - Effect of mammalian lignans on the growth of prostate cancer cell lines. AB - BACKGROUND: Mammalian lignans, enterolactone (EL) and enterodiol (ED), have been shown to inhibit breast and colon carcinoma. To date, there have been no reports of the effect of lignans on prostatic carcinoma. We investigated the effects of ED and EL on three human prostate cancer cell lines (PC-3, DU-145 and LNCaP). MATERIALS AND METHODS: Cells were treated with either 0.1% (v/v) DMSO (vehicle) or 10-100 microM of EL, ED or genistein (positive control) for 72 hours. Cell viability was measured by the propidium iodide nuclei staining fluorometric assay with each assay performed in triplicate. RESULTS: At 10-100 microM, EL significantly inhibited the growth of all cell lines, whereas ED only inhibited PC-3 and LNCaP cells. While EL was a more potent growth inhibitor than ED, both were less potent than genistein. The dose for 50% growth inhibition of LNCaP cells (IC50) by EL was 57 microM, whereas IC50 was 100 microM for ED, (the observed IC50 for genistein was 25 microM). CONCLUSION: ED and EL suppress the growth of prostate cancer cells, and may do so via hormonally-dependent and independent mechanisms. PMID- 11911283 TI - Butyric acid enhances in vivo expression of hTNF-alpha in transduced melanoma cell line. AB - Butyric acid (NaBut) and its derivatives are well-known agents eliciting tumor cell differentiation and apoptosis. In experimental models, NaBut is also used to enhance the efficacy of viral vectors. With the use of B78 murine melanoma cells transduced with the retroviral vector containing human tumor necrosis factor alpha (hTNF-alpha) gene, we investigated the ability of NaBut to increase the cytokine expression. We observed an increase in hTNF-alpha expression in vitro after incubation with NaBut. We also describe that the NaBut pro-drug tributyrin is able to increase hTNF-alpha expression in transduced B78 cells in a tumor vaccination model in mice. This observation strongly suggests a novel potential role for NaBut and its derivatives in tumor therapy. It could be used not only as a therapeutic directly acting on tumor cells but, in parallel, as a genetic vaccine "enhancer". PMID- 11911284 TI - Tissue distribution and plasma pharmacokinetics of UCN-01 at steady-state and following bolus administration in rats: influence of human alpha1-acid glycoprotein binding. AB - The primary focus of this study was to investigate the role of human alpha1-acid glycoprotein (hAGP) on the pharmacokinetics and tissue distribution of the antitumor drug UCN-01 (7-hydroxystaurosporine) in rats, following bolus administration and at steady-state blood concentration. To evaluate plasma pharmacokinetics, the rats received UCN-01 alone, UCN-01 + hAGP (87:1 ratio), or UCN-01 + hAGP (26:1 ratio) i.v. Additional rats were studied after i.m. administration of UCN-01 and i.v. administration of human AGP (87:1 ratio). For tissue distribution, rats received UCN-01 alone, UCN-01 + hAGP (87:1 ratio). One hour after drug administration, blood samples as well as various tissues and organs were collected. Plasma concentrations of UCN-01 as well as tissue accumulation were measured by HPLC using a fluorescence detector. Following i.v. bolus administration, the UCN-01 concentration-time profile declined bi exponentially. The distribution half-life was 0.2 hours, while the elimination half-life was 6.65 hours. The volume of distribution of the central compartment (Vc) was 1000 ml/kg and the volume of distribution during the elimination phase (Vdb) was 2551 ml/kg. The total body clearance (TBC) was 4.4 ml/min/kg. Co administration of hAGP with UCN-01 at 1:87 ratio did not affect the elimination half-life of UCN-01 during our sampling period, however the distribution half life was delayed by approximately 2.7-fold. Furthermore, the Vc, Vd(beta) and TBC were significantly reduced to 395 ml/kg, 735 ml/kg and 1.34 ml/min/kg, respectively. UCN-01 Vd's and TBC were reduced further by increasing human hAGP:UCN-01 ratio to 26:1. Also, hAGP administration did not significantly affect the pharmacokinetic profile of UCN-01 after i.m. administration, which was similar to that measured after i.v. administration. One hour after i.v. bolus administration, UCN-01 was distributed extensively to all tissues with a tissue/plasma ratio ranging from 10-times in the brain to more than 1000-times in the lungs. The presence of hAGP drastically reduced tissue accumulation of UCN 01, although the tissue to plasma ratio remained > 1.0 except for the brain. At steady-state blood concentration following the infusion of UCN-01 over 180 minutes, the ratio of the drug concentration to the concomitant plasma concentration remained > 1.0, even in the presence of hAGP. The data showed that the binding of UCN-01 to hAGP drastically altered its pharmacokinetics and tissue distribution, even at the plasma steady-state concentration. PMID- 11911285 TI - Quantitative measurement of telomerase catalytic subunit (hTERT) mRNA in laryngeal squamous cell carcinomas. AB - We tested 30 laryngeal squamous cell carcinomas (LSCCs) and 30 matched control laryngeal samples from the same patients for the presence of human telomerase catalytic subunit (hTERT) mRNA by using the Roche LightCycler Telo TAGGG hTERT Quantification kit. The hTERT index was calculated to express the relative quantity levels of hTERT mRNA. hTERT mRNA was detectable in 10 out of 30 (33%) laryngeal tissues covered by normal and/or reactively hyperplastic laryngeal epithelium and 23 out of 30 LSCCs (77%). The mean hTERT indices were 0.15 for control non-cancerous laryngeal samples, 0.57 for grade I, 2.35 for grade II and 3.72 for grade III LSCCs. LSCCs without detectable hTERT mRNA (23%) tended to have lower grades of disease. No correlation was found between the levels of hTERT mRNA and tumour size or locoregional lymph node status. We believe that hTERT mRNA in normal and/or reactively hyperplastic laryngeal epithelium originates from the stem cells and corresponds to the self-renewal capacity of the squamous epithelium. However, the greater quantity of h TERT mRNA in LSCCs is the result of telomerase reactivation in the process of laryngeal carcinogenesis. PMID- 11911286 TI - Proliferation characteristics of canine transmissible venereal tumor. AB - Canine transmissible venereal tumor (CTVT) grows progressively (P-phase) in the host and then spontaneously regresses (R-phase). The mechanisms behind the transition from the P-to R-phases are not well understood. In this study, in order to determine the proliferation characteristics of CTVT, we evaluated telomerase activity and enumerated nuclear organizing regions (AgNOR) and proliferating cell nuclear antigen (PCNA). It was found that CTVT cells from the P-and R-phases were both positive for telomerase activity, although it was lower in the R-phase. Evaluations of telomerase activity should take into account the stage of mitosis. Although, in the majority of cases, telomerase activity can be used to differentiate between benign and malignant tumors in dogs, other factors or markers should also be used to obtain accurate diagnoses. The PCNA-positive rate and the number and area of AgNOR per cell increased much more in the P-phase than the R-phase. However, the AgNOR values were always higher. Thus, the AgNOR count can be used to distinguish the P-and R-phases of CTVT. In addition, mitotic figures were much higher in number in the P-phase as compared to the R-phase. We believe that, during spontaneous regression of CTVT cells, slow tumor cell proliferation must contribute to the decrease in tumor size. However, shortening of tumor cell telomeres is not directly involved in this process. Other factors, such as expression of MHC antigens on CTVT cells, humoral immunity, cytokines released by the inflammatory cells and, especially, tumor infiltrating lymphocytes may contribute to CTVT regression. PMID- 11911287 TI - TAC-101, a novel retinobenzoic-acid derivative, enhances gap junctional intercellular communication among renal epithelial cells treated with renal carcinogens. AB - [4-3,5-Bis(trimethylsilyl)benzamido] benzoic acido] (TAC-101), which exhibits an anti-tumor effect, can bind to retinoic acid receptors (RARs). It has retinoid like properties, such as chemopreventive action against cancer cells. The up regulation of connexin (Cx) expression by retinoids is well known in various epithelial cells. In this study, we investigated whether TAC-101 up-regulates gap junctional intercellular communication (GJIC) in renal epithelial cells exposed to the renal carcinogens. Madin Darby canine kidney (MDCK) cells were incubated with TAC-101 for 3 days, then briefly exposed to renal carcinogens potassium bromate (KBrO3) or dimethylnitrosamine (DMN). TAC-101 increased the expression of connexin 43 protein without affecting Cx43 phosphorylation and prevented inadequate Cx43 localisation caused by KBrO3 or DMN. Consequently, TAC-101 prevented the disruption of GJIC in MDCK cells. These data suggested that TAC-101 enhanced GJIC by up-regulating Cx43 expression and that TAC-101 might be useful for the prevention of renal cell carcinoma. PMID- 11911288 TI - Anti-tumor effect associated with down-regulation of MHC class 1 antigen after co transfection of GM-CSF and IFN-gamma genes in CT26 tumor cells. AB - We previously reported that co-expression of GM-CSF and IFN-gamma genes in tumors induces a synergistic anti-tumor effect. Interestingly, we have used flow cytometry to identify two kinds of populations of CT26 cells that co-express GM CSF and IFN-gamma genes: one population (CT26/G/I-down) that expresses very low levels of MHC class I and a second population (CT26/G/I-up) that expresses high levels of H-2Kd antigen. We have compared the anti-tumor effect between CT26/I-up and CT26/G/I-down cells. When wild-type (wt) CT26 cells were injected subcutaneously into Balb/c mice, tumor was formed 5-7 days after injection. However, when both CT26/G/I-up and CT26/G/I-down cells were injected, we did not identify any tumor. The protection' study showed that both CT26/G/I-up and CT26/G/I-down. cells could induce systemic immunity against secondary challenge with unmodified parental tumor cell. CT26/G/I-down cells showed normal expression of the heavy chain of MHC class I (HC), beta2-microglobulin (beta2m) and the TAP gene. Furthermore, in CT26/G/I-down cell, although the MHC molecules were detected normally in the cytoplasmic fraction, no message was detected in the membrane fraction. Pulse-chase experiments showed that class I antigen was normally synthesized like wt CT26 or CT26/G/I-up cells. Based on our results, non MHC restricted immune effectors might play the major role in synergistic anti tumor effects with co-expression of GM-CSF and IFN-gamma in CT26 tumor model. PMID- 11911289 TI - Prognostic significance of Ape1/ref-1 subcellular localization in non-small cell lung carcinomas. AB - PURPOSE: To evaluate the prognostic value of the DNA repair/redox-protein Ape1/ref-1 in a retrospective series of consecutive non-small cell lung carcinomas (NSCLC). PATIENTS AND METHODS: Sections from 91 radically resected NSCLC were analyzed for immunohistochemical expression of Ape1/ref-1. For each case 1,000 tumor cells were evaluated to detect nuclear and cytoplasmic reactivity scored as a percentage of positive cells. With respect to sub-cellular localization and percentage of immunoreactive cells, each tumor was classified as "cytoplasmic" or "non cytoplasmic". The survival rate according to Ape1/ref-1 sub cellular localization was calculated. RESULTS: The main pattern of Ape1/ref-1 expression was nuclear. No significant difference was observed in Ape1/ref-1 pattern according to histotype (squamous vs adenocarcinoma). Among adenocarcinomas, a cytoplasmic expression of Ape1/ref-1 was significantly associated with poor survival rate in univariate (p=0.01) and multivariate (p=0.07) analyses. In addition, a cytoplasmic expression of the DNA repair protein was also predictive of worse prognosis (log-rank test, p=0.02) in cases with lymph node involvement, regardless of histotype. CONCLUSION: The results suggest a potential role of Ape1/ref-1 sub-cellular localization as a prognostic indicator in patients with NSCLC. In particular, cytoplasmic localization of the protein seems to confer a poor outcome in subgroups of patients with nodal involvement or adenocarcinoma histotype. PMID- 11911291 TI - Comparison of radiological and gross examination for detection of cancer in de fleshed skeletons. AB - The reliability of visual examination of de-fleshed bones was assessed for detection of postcranial metastatic disease in individuals known to have had cancer. This was compared with standard clinical radiological techniques. The skeletons of 128 diagnosed cancer patients from an early 20th century autopsied skeletal collection (Hamann-Todd Collection) were examined. Radiological examination detected evidence of metastatic disease in 33 individuals, compared to 11 by visual examination of the postcranial skeletons. Four of these cases were detected by both techniques. Blastic lesions were most commonly overlooked on visual examination, because they were localized to trabecular (internal bone) structures. The ilium was the most commonly affected bone, with lytic or blastic lesions detected in 30 out of 33 individuals. While the proximal femur was affected in only nine individuals, X-ray of the proximal femur and ilium detected all individuals with postcranial evidence of metastatic disease. Skeletal distribution of metastases provides no clue to the location of origin or histological subtype of the cancer. A survey of archeological human remains for metastatic cancer requires radiological examination. Such skeletal surveys should X-ray at least the ilia and femora. PMID- 11911290 TI - Serum dehydroepiandrosterone and dehydroepiandrosterone sulfate and risk of melanoma or squamous cell carcinoma of the skin. AB - BACKGROUND: Dehydroepiandrosterone (DHEA) and its analogs have potent chemoprotective actions in mouse skin tumorigenesis models. To assess this association in humans, we investigated the relationship of prediagnostic serum concentrations of DHEA and dehydroepiandrosterone sulfate (DHEAS) to the subsequent risk of developing malignant melanoma and squamous cell carcinoma of the skin in residents of Washington County, Maryland, USA. PATIENTS AND METHODS: In a nested case-control study, serum that had been stored in 1974 was thawed and assayed for DHEA and DHEAS for 23 cases of malignant melanoma and 28 cases of squamous cell carcinoma and 1-2 matched controls per case. RESULTS: The mean serum concentrations of DHEA or DHEAS were similar in cases and controls. There were no statistically significant trends in the risk of developing malignant melanoma or squamous cell skin cancer by concentration of either steroid (all p for-trends >0.30). CONCLUSION: The results of this study do not support the hypothesis that physiological concentrations of DHEA or DHEAS protect against skin cancer in humans. PMID- 11911292 TI - Clinical usefulness of chemotherapy based on an in vitro chemosensitivity test in urothelial cancer patients. AB - BACKGROUND: We evaluated the clinical usefulness of individualized chemotherapy based on an in vitro chemosensitivity test, i.e., the histoculture drug response assay (HDRA), for urothelial cancer. MATERIALS AND METHODS: 62 clinically obtained cancer specimens were studied. Each specimen was tested for sensitivity to nine anticancer drugs. The antitumor effect was calculated as the inhibition rate to their control values. The HDRA effective cytotoxic drugs were selected for clinical treatment. RESULTS: HDRA was possible in 58 out of 62 specimens (93.5%). Their chemosensitivity showed a wide variety even among the same histological category. No correlation was seen between histological grade and chemosensitivity. The effect of chemotherapy on the measurable lesions was studied in 12 patients and good clinical responses were obtained in 7 of them (58.3%). All 7 responders were the patients who received drugs predicted to be effective by HDRA. In 8 out of the 12 patients (66.7%), including 7 true-positive and 1 true-negative, HDRA correctly predlicted the clinical effect of chemotherapy. CONCLUSION: The HDRA might be feasible for predicting the efficacy and, therefore, selecting the proper anticancer drug for individual patients. PMID- 11911293 TI - Clinical estimation of the growth rate of lung cancer. AB - It is well known that the growth rate of lung tumors is closely related to prognosis and is an important determinant of responsiveness to therapy and curability. In this study, the velocity of tumor growth was calculated by dividing the area of the lesion at presentation divided by the time elapsed since symptoms were first noted. This parameter was applied to a group of patients with lung cancer and the predictive value of the velocity of tumor growth was assessed. Survival expectancy was found to be closely related to the growth rate of the tumors. The median survival time of patients with more slowly growing tumors was 102 weeks, while that of patients with fast-growing tumors was 30 weeks (log-rank test, p=0.00001). Linear regression analysis between velocity of tumor growth and tumor cell proliferation as measured by the PCNA-labelling index revealed a significant correlation between these two parameters. In conclusion, the velocity of a tumor measured in this way is an independent and significant prognostic factor for patients with lung cancer and may be used to non-invasively assess lung cancer proliferation in vivo, identifying rapidly growing tumors with poor prognosis that could benefit from a more aggressive therapy. PMID- 11911294 TI - Maximum Ki-67 staining in prostate cancer provides independent prognostic information after radical prostatectomy. AB - BACKGROUND: We have evaluated the prognostic value of Ki-67 growth fraction after radical prostatectomy, especially focusing on intermediate grade carcinomas. MATERIALS AND METHODS: 104 patients treated by radical prostatectomy for clinically localized prostate cancer were studied. The area of highest tumour grade was selected from the prostatectomy specimens and used for Ki-67 immunostaining The fraction of Ki-67 positive tumour nuclei in the area of most intense proliferation ("hot spot") was estimated, and related to biochemical failure. RESULTS: Ki-67 expression (median 6.7%, range 1.2-42.6%) was significantly associated with WHO histological grade. In univariate analysis of all 104 carcinomas, Ki-67 expression was associated with time to biochemical failure as were age, tumour dimension, WHO histological grade, pathological stage, positive surgical margins and pre-operative s-PSA. In multivariate Cox' analysis, Ki-67 expression, pathological stage and pre-operative s-PSA remained as independent predictors of time to biochemical failure. Ki-67 expression (HR 4.8, p < 0.001) was also found to be an independent predictor among moderately differentiated carcinomas. CONCLUSION: Estimates of Ki-67 growth fraction in areas of highest tumour grade may prove to be a useful prognostic biomarker after radical prostatectomy. PMID- 11911295 TI - Symptom assessment in advanced palliative home care for cancer patients using the ESAS: clinical aspects. AB - Four hundred and thirty-one cancer patients were assessed with the ESAS and a VAS QoL at admission to Hospital-based home care (HBHC) and subsequently. RESULTS: Pain and nausea were well-controlled (mean 2.5 and 1.8) whereas patients were less satisfied with appetite, activity and sense of well-being. Dyspnoea and anxiety (lung cancer, p<0.001 and p<0.01) and pain (prostate cancer, p<0.01), were related to diagnosis while activity, drowsiness, appetite and well-being to survival (p<0.05 to p<0.001). The correlations between individual symptoms and well-being were low (0.2-0.5), whereas the correlation between well-being and the Symptom Distress Score (SDS) was 0.76. "Well-being" was a better word to use than QoL. DISCUSSION: ESAS is useful in HBHC and data show that symptoms other than merely pain and nausea are of importance. As the global measurement (one VAS) of well-being has a high correlation with SDS, this single measurement may be clinically adequate for quality assurance of symptom control in dying cancer patients. PMID- 11911296 TI - Acquisition of multidrug resistance in recurrent breast cancer demonstrated by the histoculture drug response assay. AB - Recurrent breast cancer has a very poor response rate to chemotherapy. To understand the degree of acquisition of multidrug resistance in recurrent disease, 24 recurrent breast tumors and 127 primary tumors were evaluated and compared for chemosensitivity in the histoculture drug response assay (HDRA). The evaluation rate was 98.8%. The HDRA utilizes 3-dimensional culture of human tumors on collagen-gel rafts. Doxorubicin (DXR), 5-fluorouracil (5-FU) and mitomycin C (MMC) were tested as standard agents and cisplatin (CDDP) as a candidate agent on surgical specimen of breast cancer in the HDRA. In vitro drug exposure in the HDRA was for 7 days. At the end of the assay, tumor response was assessed by the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The mean inhibition rates of primary tumors vs. recurrent tumors were 57.9% and 38.6% for DXR (p<0.0005); 59.9% and 42.8% for MMC (p<0.01); 49.0% and 33.4% for 5-FU (p<0.01); and 34.5% and 16.0% for CDDP (p<0.005), respectively. The recurrent cases were pretreated clinically with CAF (cyclophosphamide, DXR and 5-FU), CEF (cyclophosphamide, epirubicin and 5-FU) or CMF (cyclophosphamide, methotrexate and 5-FU). In the CAF and CEF group, the HDRA sensitivity to CDDP was significantly lower in recurrent disease (p<0.005) than that of primary breast cancer suggesting that one agent can induce resistance to another. This is further suggested by the fact that 64.7% of the recurrent cases were resistant to all 4 agents tested as opposed to 27% of the primary cases and that only 5.9% of the recurrent cases were sensitive to three or more agents as opposed to 18% of the primary cases. The correlation of the HDRA results to clinical outcome in the study was 80.0% with 15 cases evaluated consisting of 5 true positives, 3 false positives, 7 true negatives and no false negatives. Thus, the HDRA gives useful clinical information, in particular for the specific individualized treatment design necessary to overcome the multidrug resistance problem of recurrent breast cancer. PMID- 11911297 TI - Levels of angiogenic peptides in sera from patients with carcinoid tumours during alpha-interferon treatment. AB - BACKGROUND: Alpha-interferon, a known inhibitor of angiogenesis and cell proliferation, is used in the standard treatment of patients with carcinoid tumors. We studied the levels of two angiogenic peptides (bFGF and VEGF) in sera from patients with carcinoid tumours before and during treatment with alpha interferon. The aim was to investigate if the antitumoral effect of alpha interferon in these patients could be at least in part explained by a reduction in the measured angiogenetic peptides. PATIENTS AND METHODS: Sera from 29 patients with carcinoid tumours were collected before and during alpha-interferon treatment and analyzed using commercially available ELISA-kits. RESULTS: Interferon alpha treatment did not cause reduction of bFGF and VEGF levels in serum from patients with carcinoid tumours. In fact there was no correlation between changes in bFGF or VEGF levels and treatment effect. CONCLUSION: The action of alpha-interferon does not seem to be mediated by bFGF or VEGF in patients with carcinoid tumours. If alpha-interferon has an anti-angiogenic effect in this patient group, it is probably mediated by angiogenic peptides other than bFGF and VEGF. PMID- 11911298 TI - Solitary fibrous tumor of the inguinal region: a clinicopathological, light microscopic, immunohistochemical, electron microscopic and flow-cytometric DNA study. AB - Solitary fibrous tumors (SFTs) are rare neoplasms with a probable mesenchymal origin that were first reported in the pleura but can occur in different sites. We report a case of SFT arising in the inguinal region of a 55-year-old woman. The patient presented with a mass in the left groin; she underwent wide excision of the lesion which was well-circumscribed and without evidence of adjacent soft tissue involvement. The histological, immunohistochemical and electron microscopic criteria for SFT were found. She had an uneventful recovery and she is alive without evidence of disease five years after operation. To our knowledge, this neoplasm has never been reported in this location. PMID- 11911299 TI - Chemoradiation for patients with esophageal cancer aged 80 and older. AB - BACKGROUND: The objectives of this study were to evaluate the efficacy and toxicity of concurrent chemoradiation in patients with esophageal cancer aged 80 and older. PATIENTS AND METHODS: Seven patients with esophageal cancer, aged 80 or more were treated with chemoradiation. Five received a systemic combination of cisplatin and 5-fluorouracil while 2 received daily 5-fluorouracil, concurrent with radiotherapy. The total doses of radiotherapy ranged from 50 to 65 Gy. RESULTS: Complete response was obtained in 3 patients, and partial response and no change in 2 cases each. Esophageal passage improved in 4 patients. The treatment was well-tolerated. There was no death attributable to any adverse treatment effects. None of the patients experienced grade 3 or worse acute toxicities. CONCLUSION: This study demonstrated that advanced age per se is not a sufficient reason to exclude elderly patients from aggressive treatment. PMID- 11911300 TI - Eleven cases of intraosseous lipoma of the calcaneus. AB - Intraosseous lipomas are rare benign primary bone tumors, with an incidence of one per thousand bone tumors. We studied eleven cases of intraosseous lipoma of the calcaneus. All the patients received radiographic examinations and MRI with T1-and T2-weighted images with Gd-DTPA enhancement. Seven patients received tumor curettage. Plain radiographs demonstrated that all of the lesions showed clear osteolysis surrounded by a sclerotic margin in the central body of the calcaneus. MRI revealed that all the lesions had demarcated homogenous high signal intensity lesions on both T1- and T2-weighted images, which strongly suggested adipose tissue. In 5 cases, the center of the lesion showed low signal intensity lesions on T1-weighted images, suggesting the existence of central necrosis or cyst formation. After excising the tumors of 7 patients, there was no local tumor recurrence. Surgical intervention was considered unnecessary in all cases after the diagnosis of an intraosseous lipoma was determined by MRI. PMID- 11911301 TI - Predictive value of different prognostic factors in breast cancer recurrences: multivariate analysis using a logistic regression model. AB - BACKGROUND: The aim of this study was to compare the sensitivity of different pre operative parameters in patients with breast cancer (BC) recurrence using univariate and multivariate analysis. MATERIALS AND METHODS: We retrospectively analyzed a series of 387 women (median age 60 years, range 35-83 years) who underwent curative surgery for pT1-2 BC. The patients were divided into two groups: Group 1: 325 (84.0%) patients with no evidence of disease during a median follow-up of 53 months (range 25-149 months) and Group 2: 62 (16.0%) patients who developed local or distant recurrences. RESULTS: Univariate analysis showed significant (p<0.01) differences between the two Groups in age, size and grading of the tumor and hormone receptor rate. MIB1 proliferation rate, serum markers CEA and CA 15-3, and lymph node status were not useful in predicting relapse. Multivariate analysis using a logistic regression model showed that only age, size of the tumor and hormone receptor rate independently correlate with the onset of recurrences. CONCLUSION: There is no clear correlation between BC recurrence and the majority of the prognostic factors available. Multivariate analysis of several pre-operative parameters may help to correctly select the high risk population. PMID- 11911302 TI - Detection of esophageal carcinoma using single-photon emission computed tomography of thallium-201: a preliminary report. AB - The aim of this preliminary study was to evaluate the potential usefulness of thallium-201 (Tl-201) in detecting esophageal carcinoma. A total of 15 patients with esophageal carcinoma were arranged for Tl-201 single-photon emission computed tomography (SPECT) of the chest. Thirteen of the patients had epidermoid carcinoma and two had adenocarcinoma. Meanwhile, 7 normal controls without any history of esophageal disease also accepted Tl-201 SPECT of the chest for comparison. From the 15 patients, Tl-201 chest SPECT detected esophageal carcinoma in 13 (86.7%) but not in 2 (13.3%) patients with epidermoid carcinoma. In contrast, all 7 normal controls (100.0%) had negative results of Tl-201 chest SPECT. Our study showed that the Tl-201 chest SPECT scan could be considered as a non-invasive useful imaging method in detection of esophageal carcinoma. PMID- 11911303 TI - New diagnostic approach to intracystic lesions of the breast by fiberoptic ductoscopy. AB - Intracystic tumors of the breast are uncommon and, at the time of ultrasonography and aspiration cytology, it is difficult to distinguish cancer from a benign tumor. The Fiberoptic Ductoscopy System (FDS) is an emerging technique allowing direct visual access to the ductal system of the breast. FDS was inserted through the cannulae into the cavity and we observed the intracystic tumors (3 intracystic papillomas and 2 intracytsic papillary carcinomas). The appearance of the malignant tumors was irregular, rough-shaped and they tended to bleed. On the contrary, benign tumors had smooth surfaces without bleeding. Cytological findings showed malignant cells in one out of two breast cancer patients. In addition, in the immunohistochemical study of resected tumor tissues from 5 patients, we observed positive reactions with anti-ErbB-2 antibody in 2 intracystic papillary carcinomas. In contrast, none of the histologically confirmed benign lesions (3 intracystic papillomas) gave positive results. In conclusion, the use of FDS as a non-invasive technique may provide valuable information. PMID- 11911304 TI - Long-term local hyperthermia in the treatment of advanced breast cancer (case report). AB - BACKGROUND: It is difficult to control non-resectable locally advanced primary and recurrent breast cancer by conventional modalities. Recently, hyperthermia (HT) has been recognized as an effective adjuvant to radiotherapy (RT) and chemotherapy (CT) in treatment of various malignancies, including breast cancer. PATIENT AND METHODS: The patient was a 58-year-old female Japanese, with breast cancer, T4N2M0, stage IIIb (papillo-tubular carcinoma). Previous treatment included RT and neoadjuvant CT Local HT was performed with a total number of 87 sessions given over 12 months. The mean time of each session was 40 minutes. Elevation of temperature to a tumoricidal level of 43 degrees C was confirmed. The patient received cyclophosphamide (50 mg p.o./day) and tamoxifen (20 mg p.o./day) during the whole period of HT. Due to the decreased amount of WBC, further CT was not possible, except for one course of CMF performed 3 months after the start of HT. RESULTS: The patient had a decrease in the intensity of pain even after the first 3 sessions. In one month, movement in the right shoulder became possible in an anterio-posterior direction. By 5 months, the healing of ulceration became evident. At present, the patient is in continuous CR for 15 months after HT. The movement in the shoulder joint is markedly improved in all directions. In addition, HT did not cause any notable complications. CONCLUSION: Long-term HT may be useful in the management of locally advanced breast cancer and these results should encourage further clinical study. PMID- 11911305 TI - Prognostic value of CD44 expression in colorectal carcinomas. AB - BACKGROUND: CD44 has diverse functions in cell-cell and cell-matrix interactions and its expression appears to be an indicator of invasive and metastatic behaviour in carcinomas. However, contradictory data have been reported about the correlation between CD44 expression and prognosis in colorectal carcinomas. We aimed (i) to establish whether immunohistochemically detectable CD44 expression is related to tumor aggressiveness, (ii) to correlate CD44 expression with the degree of tumor differentiation and (iii) to determine the relationship between CD44 expression and patient survival and other conventional clinicopathological features. PATIENTS AND METHODS: The immunohistochemical expression of CD44 in a series of 111 colorectal carcinomas was examined using the monoclonal mouse anti human phagocytic glycoprotein-1, CD44 (clone DF 1485) in correlation with clinicopathological variables. To achieve a reliable semi-quantitative evaluation, not only the staining intensity but also the distribution of positive tumor cells were analyzed. RESULTS: CD44 staining was high-grade positive in 42 and low-grade positive/negative in 69 tumor tissues. There was no association between CD44 expression and tumor size, histological differentiation, depth of invasion, lymph node involvement, clinical stage of the disease, or the radicality of surgical resection. CD44 expression was not correlated significantly with recurrence and distant metastases. Multivariate analysis showed that only the modified Astler-Coller (MAC) staging system was an independent prognostic factor of recurrence (HR=15.267; 15.267-6.808, 95% CI; p=0.001) and survival (HR=37.064; 13.309-103.220, 95% CI; p=0.001). Kaplan-Meier curves showed that there was no significant association between CD44 expression and recurrence and overall survival in either MAC B or C colorectal cancer. CONCLUSION: Expression of CD44 was not associated with any conventional clinicopathological features. CD44 cannot be considered as a prognostic predictor of recurrence, metastasis and overall survival. PMID- 11911306 TI - Expression patterns of alpha-, beta- and gamma-catenin in pancreatic cancer: correlation with E-cadherin expression, pathological features and prognosis. AB - BACKGROUND: The E-cadherin-catenin cell adhesion complex has been implicated in tumour invasion and metastasis. In this study, we evaluated the clinical significance of E-cadherin and catenin expression in pancreatic cancer. MATERIALS AND METHODS: The immunohistochemical expression and cellular co-localization of alpha-, beta- and gamma-catenin were investigated in 43 paraffin-embedded specimens of pancreatic cancer. The relationship between their expression and E cadherin expression, clinicopathological features and prognosis was evaluated. RESULTS: Abnormal alpha-, beta- and gamma-catenin expression was found in 37%, 44% and 40% of cases, respectively. Both alpha-catenin and gamma-catenin expression correlated with disease stage and with lymph node and distant metastases, whereas aberrant beta-catenin expression only correlated with the presence of lymph node metastases. There was a significant and progressive concordance between E-cadherin and alpha-, beta- and gamma-catenin expression patterns, respectively. The expressions of alpha-, beta- and gamma-catenin also significantly correlated with one another. All three catenins, like E-cadherin, were associated with a poor prognosis, but only E-cadherin and alpha-catenin were independent prognostic factors for cancer-specific survival. CONCLUSION: Changes in catenin expression and pancreatic cancer progression are possibly related events. The expression of E-cadherin and alpha-catenin may add useful new prognostic information. PMID- 11911308 TI - Serum p53 autoantibodies as prognostic marker in patients with oesophageal carcinoma. AB - BACKGROUND: Oesophageal carcinoma is one of the more aggressive cancers and the patients usually seek medical attention only when the disease is already advanced. Therefore it is important to have a tool, which is simple and fast, to measure and to predict the prognosis for these patients. Mutations in the p53 gene are among the most common genetic abnormalities in oesophageal carcinoma. The present study is the first study, to our knowledge, in which the relationship between the presence of p53 autoantibodies in the serum and survival has been investigated in patients with oesophageal carcinoma. PATIENTS AND METHODS: Serum from patients with oesophagal carcinoma was collected between 1996 and 1999 at the Department of Oncology, Uppsala University Hospital, Sweden. The serum samples were analysed for the presence of p53 autoantibodies using a sandwich ELISA. RESULTS: In a multivariate analysis, the presence of p53 autoantibodies was associated with decreased survival (p=0.047). Patients with extensive disease had a poor prognosis and time to death was decreased in these patients (p=0.000022). The one-year survival was 0% for these patients if they had p53 autoantibodies compared to 36% for patients with no p53 autoantibodies and extensive disease. CONCLUSION: We conclude that the presence of serum p53 autoantibodies is associated with decreased survival for patients with oesophageal carcinoma. PMID- 11911307 TI - Chemotherapy with vinorelbine, cisplatin and continuous infusion of 5 fluorouracil in locally advanced breast cancer: a promising low-toxic regimen. AB - Primary chemotherapy for locally advanced breast cancer, usually an anthracycline containing regimen, improves local disease control allowing for an initially inoperable tumour to be resected. The feasibility and efficacy of a regimen containing vinorelbine (V), cisplatin (P) and 5-fluorouracil (5-Fu) as continuous infusion (ViFuP regimen) for patients with locally advanced breast cancer were evaluated. Twenty-six patients with a T4 breast cancer presentation (eight also had synchronous distant metastases) were treated with V (20 mg total dose i.v. on day 1 and day 3), P (60 mg/m2 i.v. on day 1) and 5-Fu (200 mg/m2/d as continuous infusion) all given every 3 weeks for a maximum of 6 courses. Eleven patients had an inflammatory breast lesion, 4 had a T4a and 11 a T4b presentation. Among those with metastases, 6 had one site and 2 had two sites of disease. After chemotherapy all tumors except one became operable. Objective response was observed in 19 out of the 26 evaluable patients (73%; 95% CI: 52-88%): fourteen had a partial response (54%); 5 had a clinically complete response (19%) and 5 had complete pathological response (20%; 95% CI: 7-41%). Seven patients had stable disease (27%) while no disease progression under treatment occurred. Mild or moderate side-effects included neutropenia (G1-G2 in 58% and G3 in 31% of patients), anemia (G1 in 19%), nausea and/or vomiting (G1-G2 in 92% of patients), mucositis (G1-G2 in 23%), diarrhea (G1 in 19%), plantar-palmar erythema (G1 in 12%) and alopecia G1 in 27% of patients. We conclude that the ViFuP regimen is well-tolerated and its use results in a high response rate. Thus ViFuP may be considered a relevant alternative to more toxic regimens, with an acceptable response rate. Despite the lack of a formal demonstration of equal efficacy with more toxic regimens commonly applied in locally advanced breast cancer, testing new modalities or drugs might provide a more fruitful strategy for relevant therapeutic progress. PMID- 11911309 TI - In vitro chemosensitivity of human soft tissue sarcoma. AB - Chemotherapy is essential in the treatment of small round cell sarcomas. However, as yet there is no progress concerning the efficacy of chemotherapy in the treatment of other types of soft tissue tumors (STS). The Histoculture Drug Response Assay (HDRA) is an in vitro chemosensitivity test that has a high correlation with clinical response, the usefulness of which has been reported in various kinds of solid tumors. However, there has never been a report on its use in STS until now. In this study, in order to investigate the variation in chemosensitivity in STS, fresh biopsy or surgical samples of STS were tested using the HDRA method. Drug sensitivity testing by HDRA showed that two drugs, ADM and THP, had a significantly higher inhibitory rate than CDDP, IFOS, or VP-16 in the thirty-three soft tissue sarcomas tested. Depending on the morphological type, spindle cell sarcomas were sensitive to THP, which showed significantly higher inhibition rates than CDDP, IFOS, or VP-16. Small round cell sarcomas were relatively sensitive to all of the drugs tested. However the drug sensitivity of pleomorphic cell sarcoma was low except for ADM and THP, while its sensitivity to THP was higher than about 70%. However, there are numerous other soft tissue sarcomas that do not belong to these categories; drug sensitivity testing in each of them and the devising of individualized treatment strategies seems necessary to improve the therapeutic outcome. PMID- 11911310 TI - Differentiating benign and malignant pulmonary lesions with FDG-PET. AB - The purpose of this retrospective study was to evaluate the efficacy of positron emission tomography (PET) with 18F-fluoro-2-deoxyglucose (FDG) to differentiate benign from malignant pulmonary lesions. Fifty-five patients, suspected of having primary pulmonary neoplasm based on chest radiographic findings, underwent FDG PET scanning. Pathological diagnoses were obtained in 41 patients with a total of 43 pulmonary lesions. The other 14 patients (14 lesions) were followed-up clinically for at least four months. The standard uptake value (SUV) was determined in each patient. The SUV of the 15 benign and 40 malignant pulmonary lesions were 1.60+/-0.42 and 6.14+/-2.67, respectively. If SUV was > 2.50, the pulmonary lesion was considered as a malignant pulmonary lesion. FDG-PET could correctly detect 34 true-positive and 15 true-negative pulmonary lesions. However, 6 false-positive and one-false negative pulmonary lesions were misdiagnosed by FDG-PET. The sensitivity, specificity and accuracy of FDG-PET to differentiate between benign and malignant pulmonary lesions were 94%, 71% and 86%, respectively. FDG-PET can accurately detect malignant pulmonary lesions with a high sensitivity. However, false-positive FDG-PET findings caused by some inflammatory processes may decrease its specificity. PMID- 11911311 TI - Hepatectomy for metastases from non-colorectal and non-neuroendocrine tumor. AB - BACKGROUND: The role of hepatectomy for metastases from non-colorectal (NCR) and non-neuroendocrine (NNE) tumors has not been defined. We analyzed several factors of the primary tumor and liver metastases to clarify the prognostic determinants. PATIENTS AND METHODS: From 1990 to 1995, 33 patients underwent hepatectomy for metastases from NCR and NNE tumors. The primary tumors were gastric cancer in 9, biliary cancer in 7, pancreatic cancer in 6, breast cancer in 4 and miscellaneous tumor in 7 patients. RESULTS: The overall 5-year survival after hepatectomy was 12.1%. Morbidity and mortality rates were 21.2% and 9.1%, respectively. The liver metastases from breast cancer, unilateral liver metastases and curative resection of liver metastases were prognostic factors in univariate analysis. CONCLUSION: Resection of liver metastases from NCR and NNE tumors may offer prolonged survival for selected patients. PMID- 11911312 TI - Post-operative adjuvant chemotherapy for colorectal cancer with 5-fluorouracil (5 FU) infusion combined with 1-hexylcarbamoyl-5-fluorouracil (HCFU) oral administration after curative resection. AB - BACKGROUND: Although surgical resectability is an important prognostic factor, recurrences are commonly noted in advanced colorectal cancer patients, even after apparently curative surgery. Since such recurrences cannot be cured, better adjuvant chemotherapies are urgently required. PATIENTS AND METHODS: We studied the effect of post-operative chemotherapy using oral administration of 1 hexylcarbamoyl-5-fluorouracil (HCFU) with 5-fluorouracil (5-FU) infusion for curatively-resected Stage IIIa and IIIb colorectal cancers. This study was prospectively randomized and controlled and 314 (97.8%) out of 321 patients were determined to be candidates for statistical assessment. Group A and Group B received 5-FU intravenous injection at, respectively, 333 mg/m2 and 1000 mg/m2 body surface area/24 hours continuously for 72 hours beginning on post-operative day 0 and day 6, with oral HCFU 300 mg daily for 52 weeks beginning 2 weeks after surgery. RESULTS: There were no differences in overall 5-year survival or disease free survival between Group A and Group B. A retrospective subset analysis. however, suggested that the protocol of Group B tended to yield better 5-year survival (68.3%) for rectal cancer than that of Group A (58.8%). CONCLUSION: Inductive therapy with high-dose 5-FU in combination with oral HCFU appears to be beneficial as adjuvant chemotherapy for advanced rectal cancer with lymph node metastasis. PMID- 11911313 TI - Localized stage I-IE aggressive non-Hodgkin's lymphoma (NHL): results of prospective study with multimodality therapeutic approach. AB - BACKGROUND: A brief course of chemotherapy followed by radiation therapy was considered the best treatment for localized high-grade Non-Hodgkin's Lymphoma (NHL). The purpose of this study was to determine the efficacy and feasibility of a brief-course of anthracycline-based chemotherapy (CHOP) and consolidation radiation therapy (CRT) in a series of 57 consecutive patients with stage I-IE intermediate-high grade NHL. PATIENTS AND METHODS: Between January 1990 and December 1998, 57 consecutive patients, stage I=31 (55%) and stage IE=26 (45%), were treated with 3 cycles of CHOP regimen. Forty-four (77%) received a CRT and thirteen (23%) with primitive gastric and splenic NHL underwent radical surgery. Multivariate analysis was performed to evaluate age, lactate dehydrogenase (LDH), bulky, nodal versus extranodal localization, as prognostic factors of locoregional control and survival. RESULTS: After a median follow-up of 84 months (range 4-128 months) the 5-year overall survival (OS), disease-free survival (DFS) and event-free survival (EFS) rates were 88%, 87.5% and 84%, respectively. Risk factor analysis revealed that the LDH value was the most important adverse prognostic factor for OS and EFS. No differences were found regarding the age and or extranodal localization. The 5-year OS, DFS and EFS was 100% in thirteen patients with primitive gastric or splenic NHL treated with a radical surgical approach followed by chemotherapy without CRT. CONCLUSION: We confirm the efficacy and feasibility of a brief course of CHOP chemotherapy followed by CRT in localized I-IE intermediate-high grade NHL without adverse prognostic factors. Randomized studies are warranted in order to define the dose and the target volume of CRT (involved field or extended field) in this setting of patients. PMID- 11911314 TI - Merkel cell carcinoma: a case series of twelve patients and review of the literature. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare tumor of the skin. MATERIALS AND METHODS: A retrospective analysis of 12 cases of MCC diagnosed and treated over a 14-year period. RESULTS: Seven females and 5 males, with a median age of 72 years and MCC of the face (5), trunk (1) and extremities (6), were studied. The stage was I in 2 cases and II in 10. Eight patients had surgery, two surgery and chemotherapy and two chemotherapy only. The last two patients died of disease after 10 and 16 months. Three patients suffered locoregional recurrence after 4, 6 and 24 months and three were lost to follow-up after 8, 9 and 24 months while disease-free. Two disease-free patients died of unrelated causes after 4 and 48 months, while three were alive and well after 24, 48 and 84 months. The three relapsing patients received chemotherapy in addition to local radiation in one case. One is alive and well, the other alive with disease and the third died of disease after 84, 107 and 10 months, respectively. CONCLUSION: MCC often recurs locally. Surgery is the treatment of choice, while radiotherapy is important for local control. Chemotherapy produces responses in the neoadjuvant setting or after relapse. PMID- 11911315 TI - Carboplatin + epirubicin +VP-16 + lenograstim followed by radiotherapy + carboplatin as radiosensitizer in limited small cell lung cancer. A multicenter phase II study. AB - A phase II trial was undertaken to test the activity and toxicity of carboplatin (300 mg/m2, i.v. day 1) + epirubicin (75 mg/m2, i.v. day 1) + VP-16 (100 mg/m2, i.v. days 1 to 3) + lenograstim (5 mcg/kg, s.c. days 6 to 15) administered every 3 weeks for 4 cycles and subsequent chest irradiation (50 Gy) + daily carboplatin (25 mg/m2) in the first-line treatment of adults affected by limited small cell lung cancer (SCLC). PATIENTS AND METHODS: A single-stage phase II design was used; the complete response (CR) rate after chest radiotherapy was the primary end-point. Twenty-three CRs were required out of 38 patients to consider the treatment worthy of further study. Prophylactic cranial irradiation (PCI) was planned in case of CR. Patients aged < or = 70 were eligible if they had limited SCLC, a performance status not worse than 2 by the ECOG scale and no prior chemotherapy or radiotherapy. RESULTS: From January 1995 to April 1999, 33 patients were enrolled; the median age was 60 years. All the patients started chemotherapy; 23 patients received chest irradiation and concurrent daily carboplatin; 11 patients also received PCI. Toxicity was generally mild. Sixteen CRs (48.5%, 95% CI: 30.8-66.5) were recorded; the objective response rate was 72.7% (95% CI: 54.5-86.7). The median time-to-progression was 7.9 months (95% CI: 6.5-10.4). The median-survival was 10.7 months (95% CI: 9.2-16.1). CONCLUSION: Induction chemotherapy with carboplatin + epirubicin + VP-16 followed by chest irradiation plus concurrent daily carboplatin is well-tolerated but not sufficiently active to warrant further study in the treatment of patients with limited SCLC. PMID- 11911316 TI - Surgical management of irradiation failures in T1-T2 squamous cell carcinoma of the glottic larynx. AB - BACKGROUND: The purpose of this study was to analyse the results of salvage surgery after failure of irradiation to control the primary T1-T2 glottic cancer. MATERIALS AND METHODS: Ninety-eight patients with T1 and T2 squamous cell cancer of the glottic larynx were treated with curative intent by radiotherapy. The tumour recurred in 22 of the 98 (22%) patients. Surgical management consisted of total and frontolateral laryngectomy. Survival rates were calculated from the date of the salvage operation. RESULTS: Two of the 22 patients refused to undergo salvage surgery and one patient had pulmonary metastasis. Of the 19 patients who underwent salvage surgery, 14 (74%) had total laryngectomy and 5 (26%) had frontolateral laryngectomy. The operations were curative in 15 (79%) of the 19 patients. The overall 5-year survival rate after surgery was 78%. CONCLUSION: Stringent follow-up of patients with irradiated T1 and T2 glottic laryngeal cancer is essential to permit a successful salvage. PMID- 11911317 TI - On the role of aging in carcinogenesis. AB - BACKGROUND: Previously we identified the age at half-total incidence (Age 1/2) for a variety of tumors. In this paper, we compare the Age 1/2 with the age- standardized incidence rate (ASR) in populations that differ a) genetically and environmentally and b) genetically but not environmentally. MATERIALS AND METHODS: From 1990-era data, we calculated 95% confidence limits about mean values for Age 1/2 and ASR for cancers of the stomach, colon, rectum, lung, female breast, corpus uterus, ovary, testis and prostate. RESULTS: In populations that differed genetically and environmentally as well as those that differed genetically but not environmentally, we observed large variations in ASR, with little or no variation in Age 1/2. CONCLUSION: The determinants of ASR are distinct from the determinants of Age 1/2 and likely to be genes that can vary between tissues of tumor origin but are common to all people. PMID- 11911318 TI - Cytokeratin expression patterns as an indicator of tumour progression in oesophageal squamous cell carcinoma. AB - Recent studies point out that cytokeratins (CKs) are involved in dynamic cell remodeling during cancer progression and particularly, CK expression patterns have been associated with invasion and metastasis. In oesophageal squamous cell cancer (ESCC), lymph node (LNN) metastasis is an important step in disease progression, invariably associated with an ominous prognosis. To assess whether specific CK expression patterns could represent reliable markers of tumor progression, a series of 111 ESCCs (59 lymph node-positive, 52-negative) derived from the high- incidence area of Linxian (Northern China), were subjected to immunohistochemical (IHC) analysis with an extensive panel of CK antibodies. Statistically significant differences were observed for CK18 (p=0.01), CK19 (p=0.04) and PKK1 (p=0.02) expression between the LNN-negative and LNN-positive ESCCs. Furthermore, significant correlation between specific CK distribution pattern and progressive disease (i.e., LNN metastasis) was evidenced. The results suggest that CK8, CK18 and CK19 expression and distribution pattern could be of predictive value as a marker of disease progression as defined by the appearance of lymph node metastases in oesophageal squamous cell cancer. PMID- 11911319 TI - Severe CPT-11-induced diarrhea in presence of FK-506 following liver transplantation for hepatocellular carcinoma. AB - BACKGROUND: The treatment of malignancies in transplanted patients has become an emerging issue. The anticancer agent CPT-11 is hydrolysed to the active metabolite SN-38 and many drugs interact with its metabolism and toxicity. PATIENT AND METHODS: We studied the clinical and pharmacological interactions between CPT-11 and FK506 in a liver transplant patient. Serial plasma samples of FK506, CPT-11, SN-38 and SN-38 glucuronide were assayed by high-performance liquid chromatography. RESULTS: While no CPT-11 toxicity was observed pre operatively, several post-operative cycles of CPT-11 were complicated with severe diarrhea. No change in FK506 plasma concentrations was noted in the presence of CPT-11 but the pharmacokinetics of CPT-11 was altered in the presence of FK506. SN-38 glucuronidation was reduced for up to 12 hours following CPT-11 infusion. This increase in plasmatic exposure to unbound SN-38 might account for diarrhea. CONCLUSION: The starting dose of CPT-11 should be reduced in FK506-treated liver transplant patients. PMID- 11911320 TI - A flexible benefits tax credit for health insurance and more. AB - This essay outlines a concept for a "flexible benefits" tax credit for expanding health insurance coverage and other purposes such as retirement savings plans (with potential withdrawals for higher education, first-home ownership, and catastrophic medical expenses). Two examples are presented. The advantages of a flexible benefits tax credit are considered in terms of efficient use of the budget surplus to help meet the varied (and changing) needs of American families, to eliminate major national gaps in health insurance and pension coverage, and to advance other objectives. If the budget surplus is used wisely, political decisionmakers could achieve health insurance coverage for most uninsured workers and children and assure a future with real economic security for American families. PMID- 11911322 TI - Targeting communities with high rates of uninsured children. AB - Data from the first two rounds of the Community Tracking Study household survey show that coverage expansions through the State Children's Health Insurance Program (SCHIP) have virtually eliminated differences across communities in children's eligibility for public or private health coverage. Nevertheless, some communities continue to have very high rates of uninsured children, in large part because of lower participation rates in public programs and higher costs for employer-sponsored coverage. Participation in SCHIP may increase in high uninsurance communities as the new programs mature, although low participation rates in public programs prior to SCHIP suggest that enrollment barriers may still be greater in such communities. PMID- 11911321 TI - Reevaluation of capitation contracting in New York and California. AB - We obtained detailed quantitative and interview data from Aetna U.S. Healthcare and six physician organizations to examine changes between 1998 and 2000 in the scope of capitation contracting and delegation of responsibility for claims payment and medical management in New York and California. The physician organizations in New York included Benchmark (Continuum), Montefiore IPA, and Lenox Hill Healthcare Network. In California they included Brown and Toland Medical Group, Monarch Healthcare, and Santa Clara County IPA. In both California, where global and shared risk capitation have been common, and New York, where they have not, we find movement to reduce the scope of prepayment and a rethinking of the delegated contractual relationship by physician organizations and health plans. This represents a departure from the 1990s, when many industry participants and analysts expected capitated and delegated relationships to spread across the nation. PMID- 11911323 TI - A bias toward action: a conversation with Leonard Schaeffer. Interview by John K. Iglehart. PMID- 11911324 TI - Tracking health care costs. AB - This paper provides an update on trends in health care costs since 1999. Although the growth rate in overall costs has been stable since 1999, the trend in costs for hospital services rose, while that for prescription drugs declined, although it remains extremely high. Increased growth in hospital costs reflects the retreat from tightly managed care and labor shortages. The discrepancy between premium trends and cost trends has increased, which reflects the health insurance underwriting cycle. If these trends continue, likely responses by employers would lead to consumers' facing higher out-of-pocket costs and an increase in the number of uninsured persons. PMID- 11911325 TI - Reforming Medicare: impacts on federal spending and choice of health plans. AB - The rising cost of Medicare and well-documented problems plaguing Medicare+Choice (M+C) have increased interest in "reforming" the program. To improve efficiency, most reform proposals would rely on competitive bidding to establish payments to M+C plans. At the same time, beneficiaries would be given financial incentives to select low-cost M+C plans. A major unknown is the extent to which Medicare reforms would generate federal budgetary savings. To examine this issue, we develop three illustrative Medicare reform options that differ greatly in how Medicare would establish its payments to plans. Our results highlight the fact that Medicare should expect modest savings from reforming the program. However, other goals of reform, such as establishing more efficient payments to plans, would be achieved. PMID- 11911326 TI - Medicare+Choice: doubling or disappearing? AB - Although the changes in the program created by the Balanced Budget Act are often viewed as the reason for the current instability in the Medicare+Choice (M+C) program, in fact, health plans are having difficulties in all of their markets, not just in Medicare. It may be time to reconsider the purpose of the program and to fundamentally redesign how payments are made to managed care organizations contracting with Medicare. Two alternative approaches are suggested: treating M+C like another provider type by severing the payment linkage to spending under traditional Medicare, and overhauling the program by creating a value-based purchasing orientation rewarding plans that provide higher-quality care to beneficiaries with chronic diseases. PMID- 11911327 TI - Medicare+Choice: where did the scorekeepers go wrong? PMID- 11911328 TI - Medicare managed care: preserving an option for the future. PMID- 11911329 TI - Medicare+Choice: a time for hibernation. PMID- 11911330 TI - Paying Medicare+Choice plans: the view from MedPAC. PMID- 11911331 TI - Adding quality to the health care purchasing equation. PMID- 11911333 TI - The glucose breath test: a diagnostic test for small bowel stricture(s) in Crohn's disease. AB - The aim of this study was to determine whether an indirect noninvasive indicator of proximal bacterial overgrowth, the glucose breath test, was of diagnostic value in inflammatory bowel disease. Twenty four of 71 Crohn's disease patients tested had a positive glucose breath test. No statistical conclusions could be drawn between the Crohn's disease activity index and glucose breath test status. Of patients with radiologic evidence of small bowel stricture(s), 96.0% had a positive glucose breath test, while only one of 46 negative glucose breath test patients had a stricture. The positive and negative predictive values for a positive glucose breath test as an indicator of stricture formation were 96.0% and 97.8%, respectively. This correlation was not altered in Crohn's disease patients with fistulae or status postresection of the terminal ileum. The data in ulcerative colitis were nondiagnostic. In conclusion, the glucose breath test appears to be an accurate noninvasive inexpensive diagnostic test for small bowel stricture(s) and secondary bacterial overgrowth in Crohn's disease. PMID- 11911332 TI - Role of 5-aminosalicylic acid (5-ASA) in treatment of inflammatory bowel disease: a systematic review. PMID- 11911335 TI - Intestinal Behcet's disease presenting as a massive acute lower gastrointestinal bleed. PMID- 11911334 TI - Lactobacillus plantarum 299v inhibits Escherichia coli-induced intestinal permeability. AB - The purpose of this work was to investigate whether a probiotic bacterium, Lactobacillus plantarum 299v, could affect Escherichia coli-induced passage of mannitol across the intestinal wall. Sprague-Dawley rats were pretreated for one week by either tube feeding with L. plantarum 299v twice daily, free access to L. plantarum 299v by adding the bacterium in the drinking water, or negative control receiving regular feeding. Intestinal segments were mounted in Ussing chambers and the mucosa was exposed to control medium, E. coli, and L. plantarum 299v (alone or together). [14C]Mannitol was added as a marker of intestinal permeability and samples were taken from the serosal side. E. coli exposure induced a 53% increase in mannitol passage across the intestinal wall (P < 0.05). One week of pretreatment with L. plantarum 299v in the drinking water abolished the E. coli-induced increase in permeability. Tube feeding for one week or short term addition of L. plantarum 299v in the Ussing chambers had no effect on the permeability provoked by E. coli challenge. Notably, L. plantanum 299v itself did not change the intestinal passage of mannitol. These data demonstrate that pretreatment with L. plantarum 299v, which is a probiotic bacterium, protects against E. coli-induced increase in intestinal permeability, and that L. plantarum 299v alone has no influence on the intestinal permeability. Thus, this study supports the concept that probiotics may exert beneficial effects in the gastrointestinal tract. PMID- 11911336 TI - Post-liver transplant Crohn's disease: graft tolerance but not self-tolerance? AB - Inflammatory bowel disease (IBD) rarely occurs de novo after liver transplantation, and when it does usually presents as ulcerative colitis in patients transplanted for primary sclerosing cholangitis. We present two patients who developed de novo Crohn's colitis two and three years after liver transplant for primary biliary cirrhosis (PBC) and chronic hepatitis B infection, respectively. Both were on maintenance immunosuppression with a calcineurin inhibitor and azathioprine with no evidence of allograft rejection. Investigations for enteric and opportunistic infection were negative. In the nontransplant setting, the development of IBD is likely multifactorial with an immune origin. In our cases, the immunosuppression was titrated to minimal levels when IBD developed. Clearly, although a significant degree of allograft tolerance can occur, autoimmune diseases can still develop elsewhere, suggesting selective immune tolerance. PMID- 11911337 TI - Nitric oxide level profile in human liver transplantation. AB - The aim of this study was to monitor nitric oxide blood levels at various times intraoperatively and following liver transplantation in humans. Nitric oxide production was assessed directly as circulating nitrosyl-hemoglobin adducts by electron paramagnetic resonance spectroscopy in 22 patients undergoing orthotopic liver transplantation. Two significant peaks in nitrosylhemoglobin levels were detected at 5 and 60 min after reperfusion (5.02 +/- 3.33 arbitrary units and 5.75 +/- 4.19, respectively, vs 3.33 +/- 2.28 under basal state; P < 0.05 for both comparisons). Postoperative nitrosyl-hemoglobin levels remained elevated, up to 5.42 +/- 0.89 arbitrary units (P < 0.05 vs basal values). Neither soluble intercellular adhesion molecule-1 or soluble endothelial-leukocyte adhesion molecule concentrations were altered intraoperatively. Only the former was significantly raised after transplantation. Neutrophil elastase levels showed an early increase and remained high throughout surgery, returning to basal values after transplantation. No correlations were found among studied parameters. These data suggest that nitric oxide may play a role in ischemia-reperfusion phases in human liver transplantation. Mechanisms other than leukocyte-endothelial adhesion and neutrophil activation seem to affect nitric oxide production under these conditions. PMID- 11911338 TI - Hepatitis C virus (HCV) RNA level determined by second-generation branched-DNA probe assay as predictor of response to interferon treatment in patients with chronic HCV viremia. AB - Using first- and second-generation branched-DNA probe assays (1st- and 2nd-bDNA), we investigated the predictors of favorable clinical response to interferon (IFN) treatment in patients with chronic HCV viremia. A total of 122 patients (85 genotype lb and 37 genotype 2a) with chronic HCV viremia received 24-week IFN alpha treatment. Patients with sustained clearance of serum HCV RNA by polymerase chain reaction at six months after IFN treatment were defined as having a sustained response (SR). HCV RNA level was determined by 1st- and 2nd-bDNA assays prior to treatment. Mean HCV RNA level by 1st-bDNA was significantly higher in genotype lb patients [5.4 x 10(6) HCV genome equivalent (Meq)/ ml] than in genotype 2a patients (0.9 Meq/ml) (P < 0.05). There was no significant difference between patients with these genotypes in the level by 2nd-bDNA (1b: 5.2 Meq/ml and 2a: 3.1 Meq/ml). SR was achieved by 43 (35.2%) of 122 patients. Mean HCV RNA levels by both the 1st- and 2nd-bDNA of SR patients (1.0 and 1.9 Meq/ml) were significantly lower than those of non-SR patients (5.3 and 6.0 Meq/ml) (both P < 0.05). The SR rate in genotype 2a patients (59.5%) was significant higher than in genotype lb patients (24.7%) (P < 0.05). Stepwise logistic regression analysis showed that HCV RNA level < or = 1.0 Meq/ml by 2nd-bDNA (odds ratio = 7.6, compared to level > 1.0 Meq/ml, P < 0.05) was a significant predictive cutoff for SR. Using 2nd-bDNA, a significantly higher rate of SR was found in genotype lb patients with level < or = 1.0 Meq/ml (57.6%) than in those with level > 1.0 Meq/ml (3.8%) (P < 0.05). The SR rate of genotype 2a patients with level >1.0 Meq/ml (68.6%) was somewhat higher than for those with level < or = 1.0 Meq/ml (52.4%). These findings suggested that, using 2nd-bDNA, a low HCV RNA level of < or = 1.0 Meq/ml was the most favorable marker of successful IFN treatment and that patients with genotype 2a, even those with level >1.0 Meq/ml, had a high rate of SR to IFN treatment. PMID- 11911339 TI - Thiamine deficiency in hepatitis C virus and alcohol-related liver diseases. AB - Thiamine deficiency is a common feature in chronic alcoholic patients, and its pathophysiology remains poorly understood. Until now, thiamine deficiency has been considered to be mainly the result of alcoholism irrespective of the underlying liver disease. The aims of the study were to compare the prevalence of thiamine deficiency in alcohol- and hepatitis C virus-(HCV-) related cirrhosis and in patients with chronic hepatitis C without cirrhosis. Forty patients with alcoholic cirrhosis (group A), 48 patients with HCV-related cirrhosis (group B), and 59 patients with chronic hepatitis C without cirrhosis (group C) were included prospectively. Thiamine status was evaluated by concomitant determination of erythrocyte transketolase activity, thiamine diphosphate (TDP) effect, and direct measurement of erythrocyte thiamine and its phosphate esters by HPLC. Thiamine was mainly present in erythrocytes in its diphosphorylated form. Prevalence of thiamine deficiency and levels of TDP in thiamine-deficient patients were similar in patients of group A (alcoholic cirrhosis) and of group B (viral C cirrhosis). None of the patients with chronic hepatitis (group C) was deficient. Thiamine deficiency was not correlated with the severity of the liver disease or disease activity. No impairment of thiamine phosphorylation was found in the three groups. conclusion, alcoholic or HCV-related cirrhotics have the same range of thiamine deficiency, while no patient without cirrhosis has thiamine deficiency, and impaired phosphorylation does not account for the deficiency observed in cirrhotics. We suggest that thiamine should be given to patients with cirrhosis irrespective of its cause. PMID- 11911340 TI - Various S-GOT/S-GPT ratios in nonviral liver disorders and related physical conditions and life-style. AB - The relationship between the GOT/GPT ratio in nonviral liver disorders and underlying physical condition and life-style were evaluated. The subjects were 12,808 male railway company workers who underwent an annual health checkup. Nonviral liver disorders were defined as elevated transaminases (GOT > 76 IU/liter or GPT > 86 IU/liter, while negative for hepatitis B and C markers (282 cases). Controls were 9,783 males with normal findings for GOT, GPT, and y-GTP. By logistic regression analysis, GOT-dominant liver disorders were significantly related to alcohol consumption, hypertriglyceridemia, and diabetes mellitus. They were still significant on multivariate analysis. GPT-dominant liver disorders were significantly related to obesity, less exercise, hypercholesterolemia, and hypertriglyceridemia. Obesity and hypercholesterolemia were significant on multivariate analysis. In conclusion, the relationship between hypertriglyceridemia or diabetes mellitus and GOT-dominant disorders, which was not explained empirically, could indicate another pathogenesis for nonviral liver disorders, such as underlying insulin resistance. PMID- 11911341 TI - Evidence for prostaglandin-producing supressor cells in HCV patients with normal ALT. AB - Our objective was to verify the presence of prostaglandin-producing suppressor cells in response to hepatitis C virus antigens in peripheral blood mononuclear cells proliferation. Standard proliferation tests were performed in 31 patients: 20 with chronic hepatitis C after antiviral treatment [7 long-term responders (LTR), 7 relapsers (RR), 6 nonresponders (NR)], 7 with HCV infection with persistently normal aminotransferase levels (PNAL), and 4 with hepatocellular carcinoma. Six antigens were used from the core and NS3 regions. A modified proliferation assay consisting of the addition of indomethacin was also done. Lymphoproliferative responses to the HCV antigens were detectable in 27% (11/41) of test points of LTR, 10% (3/31) of RR, 26% (9/35) of NR, and 18% (7/39) of patients with PNAL. Indomethacin only had effect in PNAL patients, by increasing the frequency of reactivity from 18% (7/39) to 36% (14/39) tests points (P = 0.037); also, in three of these patients (43%) indomethacin strongly modified proliferation to core 31-50 and NS3 1248-1261 antigens, increasing both the frequency and stimulation index from 33% (6/18) to 72% (13/18) (chi2 = 5.43, P = 0.019) and 1.89 +/- 0.43 to 6.18 +/- 4.74 (P = 0.028), respectively. These results suggest that prostaglandin-producing suppressor may play a role in chronic HCV infection by inhibiting cellular immune responses in patients with persistently normal ALT. PMID- 11911342 TI - Autoimmune thrombocytopenia induced by PEG-IFN-alpha2b plus ribavirin in hepatitis C. PMID- 11911343 TI - Lamivudine treatment for recurrent pancreatitis associated with reactivation of chronic B hepatitis. PMID- 11911344 TI - Occupational mortality from squamous cell carcinoma of the esophagus in the United States during 1991-1996. AB - The epidemiology of esophageal squamous cell cancer has remained poorly understood. The occupational distribution of this cancer may provide clues about its yet unknown etiology. Data files from the National Center for Health Statistics (NCHS) of the United States offer a unique source to study causes of death, broken down by occupation and industry. The number of deaths from esophageal cancer was retrieved from the computerized US vital statistics. Mortality by occupation or industry was expressed as standardized proportional mortality ratio (PMR), adjusted by age, gender, and ethnicity. Between 1991 and 1996, 63,717 subjects died from esophageal squamous cell carcinoma. Mortality was particularly high among nonwhites and men. The industrial and the occupational distributions shared a similar pattern. Mortality from esophageal squamous cell carcinoma occurred more frequently among subjects exposed to silica dust, such as brickmasons and stonemasons, concrete and terrazzo finishers, roofers, and construction laborers. It was also high in such industries as unspecified machinery or manufacturing and such occupations as unspecified material handlers, janitors, or cleaners. It was low in industries and occupations associated with agriculture, clergy, work in religious organizations, and textiles. In conclusion, mortality from esophageal squamous cell carcinoma appeared to be low in occupations associated with less consumption of alcohol and tobacco. It was high among occupations potentially associated with exposure to silica dust and chemical solvents or detergents. PMID- 11911345 TI - Identification of spasmolytic polypeptide expressing metaplasia (SPEM) in remnant gastric cancer and surveillance postgastrectomy biopsies. AB - Following gastrectomy, the remnant oxyntic mucosa is at increased risk of developing adenocarcinoma. Alkaline pancreaticoduodenal reflux, carcinogen production from intragastric bacterial overgrowth, denervation, and devascularization have been implicated in this malignant transformation. Recent reports have described a novel spasmolytic polypeptide (SP) expressing metaplastic lineage designated as SPEM. This lineage has been identified in the mucosa surrounding gastric adenocarcinomas, and SP staining has been observed in the cells of surface dysplasia and invasive malignancy. In this study we describe 19 cases of remnant gastric adenocarcinoma from Japan. In addition, we studied surveillance biopsies in 90 patients who underwent antrectomy for carcinoma. SPEM was identified in the mucosa surrounding 88% of the remnant cancers, as well as in 61% of the surveillance biopsies. In the malignant resections, 67% of the surface dysplasia displayed SP positive cells, and 25% revealed SP immunostaining within invasive malignant cells. These findings implicate SPEM as a potential precursor lesion of gastric adenocarcinoma. PMID- 11911347 TI - Spontaneous gas-forming liver abscess caused by Salmonella within hepatocellular carcinoma: a case report and review of the literature. AB - In conclusion, pyogenic liver abscess in hepatocellular carcinoma is unusual. Most of the reported cases occurred after a treatment such as transcatheter arterial embolization or percutaneous ultrasound-guided ethanol injection. Salomonella very rarely causes pyogenic liver abscesses. Only 14 cases have been reported in the English literature since 1911. Salmonella liver abscess occurring within a primary neoplasm is even rarer. There were only two such cases described in patients with hepatocellular carcinoma before. The present case is the third one, but it may be the first case of obvious spontaneous gas-forming liver abscess caused by Salmonella within hepatocellular carcinoma. PMID- 11911346 TI - Gastric carcinoid tumors without autoimmune gastritis in Japan: a relationship with Helicobacter pylori infection. AB - In Japan, most cases of gastric carcinoid tumor (GCT) are unassociated with either autoimmune gastritis (AIG) showing type-A chronic atrophic gastritis (CAG A) or Zollinger-Ellison syndrome (ZES). However, the pathogenesis of this tumor remains unknown. Recent studies have determined that Helicobacter pylori infection induces gastric carcinoid in Mongolian gerbils and that H. pylori lipopolysaccharide exerts a mitogenic effect on ECL cells. We examined five patients with histologically diagnosed GCT, 40 patients with H. pylori-positive gastric ulcer (Hp+GU), 24 patients with H. pylori-positive duodenal ulcer (Hp+DU), and 12 patients with AIG showing CAG-A topographically. We compared the prevalence of H. pylori infection, and the levels of gastrin and pepsinogen (PG) in the serum of patients with GCT with those of patients with Hp+GU, or Hp+DU, and AIG. We also investigated the histological characteristics of the tumor and the gastric corpus mucosa in the GCT patients. The levels of serum gastrin and PG I and II were measured using an RIA kit. In all five (100%) patients with GCT, H. pylori infection was present, without any evidence of AIG or ZES. The serum levels of gastrin in the GCT patients were higher than those in either Hp+GU or Hp+DU patients and lower than those in the AIG patients. In contrast, serum PG I levels and the PG I/II ratio were lower in the GCT group than in the Hp+GU or Hp+DU groups. Histologically, all GCTs were ECL cell tumors and peritumoral corporal mucosal atrophy was observed in four of the five patients with GCT. In conclusions, H. pylori infection and hypergastrinemia were found in the patients with GCT without AIG. This finding suggests that H. pylori infection may induce corporal mucosal atrophy and hypergastrinemia that can produce a GCT with time. PMID- 11911349 TI - Heterozygosity for factor V Leiden and G20210A prothrombin genotypes in a patient with mesenteric vein thrombosis. PMID- 11911348 TI - Rat small intestinal goblet cell kinetics in the process of restitution of surface epithelium subjected to ischemia-reperfusion injury. AB - Repair of superficial damage to gastrointestinal mucosa occurs by a process called restitution. Goblet cells reside throughout the length of the intestine and are responsible for the production of mucus. However, a kinetic analysis of goblet cell dynamics of small intestine in restitution has hitherto not been reported. The aim of the present study was to investigate the role of goblet cells in the process of restitution of rat small intestine subjected to ischemia and ischemia-reperfusion injury, and therefore intestinal epithelium from rats subjected to both ischemia and ischemia-reperfusion was studied. Detachment of enterocytes was observed after 5-min of reperfusion. After 20-30 minutes of reperfusion, the denuded villous tips were covered with goblet cells. Within 75 min of reperfusion the epithelium restitution was complete. On the other hand, restitution was not observed in ischemia group. These data suggest that goblet cells may play an important role in restitution after ischemia-reperfusion injury. PMID- 11911350 TI - Reactive oxygen species and vascular cell adhesion molecule-1 in distant organ failure following bile duct obstruction in mice. AB - Pulmonary injury with leukocyte infiltration is a frequent occurrence in obstructive cholangitis patients. We wished to evaluate the roles of reactive oxygen species and vascular cell adhesion molecule-1 (VCAM-1) in this distant organ failure. Wild type (WT) and transgenic (SODtg) mice overexpressing superoxide dismutase underwent bile duct ligation and transection (BDL). VCAM-1 expression was quantified, and histopathology was assessed for the liver and lung. BDL resulted in increased leukocyte infiltration to the lung at five days in WT mice. VCAM-1 expression significantly increased in WT mouse liver at three days and WT mouse lung at five days. When these same experiments were performed in SODtg mice, these increases in leukocyte infiltration and VCAM-1 expression in lung were significantly attenuated. These data suggest that reactive oxygen species produced in response to BDL may up-regulate VCAM-1 expression in the lung and play an important role in the pathophysiology of this pulmonary injury. PMID- 11911351 TI - Acute pancreatitis after single-dose exposure to propofol: a case report and review of literature. PMID- 11911352 TI - Epidermoid cysts of the spleen occurring in sisters. PMID- 11911353 TI - Endoscopic evaluation of patients with partial gastrectomy and iron deficiency. AB - Endoscopy is indicated for the evaluation of unexplained iron deficiency to rule out neoplasia. Iron deficiency is common in postgastrectomy patients. The endoscopic yield for significant pathology in these patients is unknown but is expected to be lower than for other iron-deficient groups. A retrospective case control study with 2:1 matching was performed comparing iron-deficient patients (ferritin < or = 50 microg/liter) having prior Billroth I or Billroth II gastrectomy to matched iron-deficient controls with normal gastric anatomy. There were 52 postgastrectomy patients and 113 controls. There were no significant differences between postgastrectomy patient and controls in age, gender ratio, or laboratory test results, with the exception of MCV (88.9 +/- -1.1 vs 86.0 +/- 0.8, mean +/- SEM, P = 0.048) There were no significant differences in the prevalence of upper gastrointestinal pathology (24.5% vs 29.2%), large (>1 cm) colon polyps (8.3% vs 5.2%), or the presence of any adenomatous colon polyp (28.6% vs 18.9%). There were no malignancies. In conclusion, prevalence of clinically significant pathology is similar for postgastrectomy and nonpostgastrectomy iron-deficient patients. Endoscopic evaluation of iron deficiency should not differ in postgastrectomy patients. PMID- 11911354 TI - Relationship between acid reflux episodes and gastroesophageal reflux symptoms is very inconstant. AB - In this prospective study 244 consecutive patients presenting with typical and chronic signs of gastroesophageal reflux were included. Conventional 24-hr esophageal pH monitoring was carried out to establish the symptom association probability, the concordance index, and the symptom sensitivity index. The symptom association probability could be calculated in 110 patients (45%). Two groups were identified: group 1 had normal duration of esophageal acid exposure; subgroup la (nonsignificant symptom association probability) included 39 patients (35.5%) and subgroup lb (significant symptom association probability) included 24 patients (21.8%); group 2 had abnormal duration of esophageal acid exposure; subgroup 2a (nonsignificant symptom association probability) included 21 patients (19.1%) and subgroup 2b (significant symptom association probability) included 26 patients (23.6%). In all, 56.6% of the patients presented typical symptoms of reflux not directly determined by one or repeated acid reflux episodes. The correlation between symptom association probability and the symptom sensitivity index allows for more accurate determination of esophageal acid sensitivity (subgroups lb and 2b). PMID- 11911355 TI - Ineffective esophageal motility is a primary motility disorder in gastroesophageal reflux disease. AB - The relationship between esophageal motor abnormalities and GERD has been widely studied. The purpose of this study was to identify the prevalence of ineffective esophageal motility (IEM) in patients with GERD. In addition, we also evaluated esophageal acid exposure, acid clearance, and endoscopic esophagitis in GERD patients with IEM. Of 89 patients enrolled in this study, 47 (52.8%) were found to have nonspecific esophageal motility disorder (NEMD). Forty-four of the 47 (93.6%) patients with NEMD met the diagnostic criteria for IEM. The overall incidence of IEM in GERD patients was 49.4%. Patients with IEM had significant increases in upright and recumbent mean fraction of time pH < 4 (6.70% and 4.38%) and mean recumbent esophageal acid clearance (12.45 min/reflux) when compared to those with other motility findings. Seventeen of the 44 (39%) IEM patients did not have endoscopic esophagitis. On the other hand, 26 of the 39 (67%) patients with normal manometry had endoscopic esophagitis. We concluded that not only is the prevalence of IEM high in GERD, but also that IEM patients have more recumbent gastroesophageal reflux and delayed acid clearance. Combined with endoscopic findings, we propose that IEM can be viewed as a specific entity of primary esophageal motility disorder in patients with GERD. PMID- 11911356 TI - Fat and esophageal sensitivity to acid. AB - The hypothesis that fat increases esophageal sensitivity to acid was tested in eight patients with gastroesophageal reflux disease and 11 healthy subjects. Protocol 1 included randomized intragastric infusions of saline or Lipofundin S 20% (306 kcal) on two separate days, followed after 30 and 90 min by an 8 ml/min intraesophageal infusion of 0.1 N HCl. The time to the onset of heartburn and the maximum heartburn score by visual analog scale during the acid infusion were similar after intragastric saline (2 min and 29.5 mm, medians) and fat (2 min and 20.5 mm). Protocol 2 included two 8 ml/min intraesophageal infusions of 0.2 N HCI diluted in an equal volume of saline or Lipofundin S 20% at a time interval of 10 min in randomized order. The time to the onset of heartburn and the maximum heartburn score were unaffected by the presence of fat in the esophageal infusate (2.5 min and 53 mm without vs 1.5 min and 49 mm with fat). We conclude that fat does not increase esophageal sensitivity to acid. PMID- 11911357 TI - Clinical relevance of cagE gene from Helicobacter pylori strains in Japan. AB - It has been reported that H. pylori-containing cagE was associated with duodenal ulcer. The aims of the present study were to clarify the association between the cagE gene and clinical outcome and to analyze the relationship between the cagE gene and two other virulence factors--cagA and vacA--in two areas in Japan (Fukui and Okinawa) where the prevalence of duodenal ulcer and gastric cancer risk are quite different. Eighty of 81 isolates possessed the cagE gene, and all isolates possessed the cagA gene. The vacA genotype s1c/ml was a major genotype in both areas in Japan. There was no significant association between cagE, cagA status, or vacA genotype and clinical outcome. Phylogenetic analysis of the cagE gene indicated that most Japanese isolates formed a different cluster from strains isolated in the West with an association with the vacA genotype. In conclusion, the strains with cagE, cagA, and the s1c/ml genotype of vacA are predominant in Japan regardless of clinical outcome and construct a different phylogenetic cluster from those in the West. PMID- 11911358 TI - Cigarette smoking promotes atrophic gastritis in Helicobacter pylori-positive subjects. AB - Aging and smoking are known to promote atrophic gastritis (AG) and intestinal metaplasia (IM). This study investigated the relationship between Helicobacter pylori (Hp) infection, aging, smoking, and AG/IM. Ninety-six Hp-negative and 231 Hp-positive subjects were divided according to age; (< or = 39 years, 40-59 years, and > or = 60 years) and smoking history (never smoked, or currently smoking). Histologic grading was performed according to the updated Sydney system. Fasting pH, total bile acid (TBA) concentration, and ammonia (NH3) concentration in gastric juice were measured. Comparisons were made based on Hp status, age, and smoking. Independent relative risks for severe AG and IM were calculated. Grades of atrophy and IM were significantly higher in Hp-positive subjects, and these increased with age. Within Hp-positive subjects, grades of atrophy and IM were higher in smokers and in the middle and upper age groups. Within Hp-positive subjects, gastric pH and TBA were similarly higher in smokers and older subjects. An increased risk of severe AG/IM was statistically associated with smoking (OR 9.31, 3.85-22.50/OR 4.91, 1.90-12.68) and high TBA concentrations (OR 2.92, 1.19-7.17/OR 3.28, 1.25-8.62). Both Hp infection and aging are closely related to the development of AG and IM. Cigarette use and high TBA concentrations may play a role in the progression of AG and IM in Hp-positive subjects. PMID- 11911359 TI - A revised model for U4atac/U6atac snRNA base pairing. PMID- 11911360 TI - Highly conserved NIKS tetrapeptide is functionally essential in eukaryotic translation termination factor eRF1. AB - Class-1 polypeptide chain release factors (RFs) play a key role in translation termination. Eukaryotic (eRF1) and archaeal class-1 RFs possess a highly conserved Asn-Ile-Lys-Ser (NIKS) tetrapeptide located at the N-terminal domain of human eRF1. In the three-dimensional structure, NIKS forms a loop between helices. The universal occurrence and exposed nature of this motif provoke the appearance of hypotheses postulating an essential role of this tetrapeptide in stop codon recognition and ribosome binding. To approach this problem experimentally, site-directed mutagenesis of the NIKS (positions 61-64) in human eRF1 and adjacent amino acids has been applied followed by determination of release activity and ribosome-binding capacity of mutants. Substitutions of Asn61 and Ile62 residues of the NIKS cause a decrease in the ability of eRF1 mutants to promote termination reaction in vitro, but to a different extent depending on the stop codon specificity, position, and nature of the substituting residues. This observation points to a possibility that Asn-Ile dipeptide modulates the specific recognition of the stop codons by eRF1. Some replacements at positions 60, 63, and 64 cause a negligible (if any) effect in contrast to what has been deduced from some current hypotheses predicting the structure of the termination codon recognition site in eRF1. Reduction in ribosome binding revealed for Ile62, Ser64, Arg65, and Arg68 mutants argues in favor of the essential role played by the right part of the NIKS loop in interaction with the ribosome, most probably with ribosomal RNA. PMID- 11911361 TI - Mutations in RRM4 uncouple the splicing repression and RNA-binding activities of polypyrimidine tract binding protein. AB - The polypyrimidine tract binding protein (PTB, or hnRNP I) contains four RNA binding domains of the ribonucleoprotein fold type (RRMs 1, 2, 3, and 4), and mediates the negative regulation of alternative splicing through sequence specific binding to intronic splicing repressor elements. To assess the roles of individual RRM domains in splicing repression, a neural-specific splicing extract was used to screen for loss-of-function mutations that fail to switch splicing from the neural to nonneural pathway. These results show that three RRMs are sufficient for wild-type RNA binding and splicing repression activity, provided that RRM4 is intact. Surprisingly, the deletion of RRM4, or as few as 12 RRM4 residues, effectively uncouples these functions. Such an uncoupling phenotype is unique to RRM4, and suggests a possible regulatory role for this domain either in mediating specific RNA contacts, and/or contacts with putative splicing corepressors. Evidence of a role for RRM4 in anchoring PTB binding adjacent to the branch site is shown by mobility shift and RNA footprinting assays. PMID- 11911362 TI - Saccharomyces cerevisiae nucleolar protein Nop7p is necessary for biogenesis of 60S ribosomal subunits. AB - To identify new gene products that participate in ribosome biogenesis, we carried out a screen for mutations that result in lethality in combination with mutations in DRS1, a Saccharomyces cerevisiae nucleolar DEAD-box protein required for synthesis of 60S ribosomal subunits. We identified the gene NOP7that encodes an essential protein. The temperature-sensitive nop7-1 mutation or metabolic depletion of Nop7p results in a deficiency of 60S ribosomal subunits and accumulation of halfmer polyribosomes. Analysis of pre-rRNA processing indicates that nop7 mutants exhibit a delay in processing of 27S pre-rRNA to mature 25S rRNA and decreased accumulation of 25S rRNA. Thus Nop7p, like Drs1p, is required for essential steps leading to synthesis of 60S ribosomal subunits. In addition, inactivation or depletion of Nop7p also affects processing at the A0, A1, and A2 sites, which may result from the association of Nop7p with 35S pre-rRNA in 90S pre-rRNPs. Nop7p is localized primarily in the nucleolus, where most steps in ribosome assembly occur. Nop7p is homologous to the zebrafish pescadillo protein necessary for embryonic development. The Nop7 protein contains the BRCT motif, a protein-protein interaction domain through which, for example, the human BRCA1 protein interacts with RNA helicase A. PMID- 11911363 TI - Defining a 5' splice site by functional selection in the presence and absence of U1 snRNA 5' end. AB - Pre-mRNA splicing in metazoans is mainly specified by sequences at the termini of introns. We have selected functional 5' splice sites from randomized intron sequences through repetitive rounds of in vitro splicing in HeLa cell nuclear extract. The consensus sequence obtained after one round of selection in normal extract closely resembled the consensus of natural occurring 5' splice sites, suggesting that the selection pressures in vitro and in vivo are similar. After three rounds of selection under competitive splicing conditions, the base pairing potential to the U1 snRNA increased, yielding a G100%U100%R94%A67%G89%U76%R83% intronic consensus sequence. Surprisingly, a nearly identical consensus sequence was obtained when the selection was performed in nuclear extract containing U1 snRNA with a deleted 5' end, suggesting that other factors than the U1 snRNA are involved in 5' splice site recognition. The importance of a consecutive complementarity between the 5' splice site and the U1 snRNA was analyzed systematically in the natural range for in vitro splicing efficiency and complex formation. Extended complementarity was inhibitory to splicing at a late step in spliceosome assembly when pre-mRNA substrates were incubated in normal extract, but favorable for splicing under competitive splicing conditions or in the presence of truncated U1 snRNA where transition from complex A to complex B occurred more rapidly. This suggests that stable U1 snRNA binding is advantageous for assembly of commitment complexes, but inhibitory for the entry of the U4/U6.U5 tri-snRNP, probably due to a delayed release of the U1 snRNP. PMID- 11911364 TI - Both ran and importins have the ability to function as nuclear mRNA export factors. AB - The Ran protein regulates nucleocytoplasmic transport mediated by the karyopherin family of nuclear transport factors. Ran is converted to the active, GTP bound form in the nucleus and then binds to a conserved domain found in all karyopherins. This interaction induces cargo binding for exportins and cargo release for importins. In either case, the Ran.GTP is then transported to the cytoplasm by the karyopherin, where it is hydrolyzed to Ran.GDP. To ask whether Ran could function as a nuclear mRNA export factor, we fused Ran to the MS2 coat protein and inserted MS2 RNA-binding sites into an unspliced cat mRNA that is normally sequestered in the nucleus. Coexpression of MS2-Ran induced cat mRNA export and CAT enzyme expression as effectively as, for example, an MS2-Rev fusion protein. MS2-Ran dependent nuclear mRNA export was reduced by inhibitors specific for Crm1, but not blocked as was seen with MS2-Rev. Consistent with the hypothesis that Crm1 is not the only karyopherin cofactor for MS2-Ran mediated mRNA export, we show that not only Crm1 but also CAS, transportin, importin beta and exportin t can all export mRNA from the nucleus when tethered via the MS2 RNA binding domain. In contrast, two shuttling hnRNPs, hnRNP A1 and hnRNP K, proved unable to function as nuclear RNA export factors when expressed as MS2 fusions. Together, these data argue that karyopherins that normally function to transport proteins into or out of the nucleus are also capable of exporting tethered mRNA molecules. PMID- 11911365 TI - Determinants of the recognition of enteroviral cloverleaf RNA by coxsackievirus B3 proteinase 3C. AB - The initiation of enteroviral positive-strand RNA synthesis requires the presence of a functional ribonucleoprotein complex containing a cloverleaf-like RNA secondary structure at the 5' end of the viral genome. Other components of the ribonucleoprotein complex are the viral 3CD proteinase (the precursor protein of the 3C proteinase and the 3D polymerase), the viral 3AB protein and the cellular poly(rC)-binding protein 2. For a molecular characterization of the RNA-binding properties of the enteroviral proteinase, the 3C proteinase of coxsackievirus B3 (CVB3) was bacterially expressed and purified. The recombinant protein is proteolytically active and forms a stable complex with in vitro-transcribed cloverleaf RNA of CVB3. The formation of stable complexes is also demonstrated with cloverleaf RNA of poliovirus (PV) 1, the first cloverleaf of bovine enterovirus (BEV) 1, and human rhinovirus (HRV) 2 but not with cloverleaf RNA of HRV14 and the second cloverleaf of BEV1. The apparent dissociation constants of the protein:RNA complexes range from approx. 1.7 to 4.6 microM. An electrophoretic mobility shift assay with subdomain D of the CVB3 cloverleaf demonstrates that this RNA is sufficient to bind the CVB3 3C proteinase. Binding assays using mutated versions of CVB3 and HRV14 cloverleaf RNAs suggest that the presence of structural features rather than a defined sequence motif of loop D are important for 3C proteinase-RNA interaction. PMID- 11911366 TI - Posttranscriptional modifications in the A-loop of 23S rRNAs from selected archaea and eubacteria. AB - Posttranscriptional modifications were mapped in helices 90-92 of 23S rRNA from the following phylogenetically diverse organisms: Haloarcula marismortui, Sulfolobus acidocaldarius, Bacillus subtilis, and Bacillus stearothermophilus. Helix 92 is a component of the ribosomal A-site, which contacts the aminoacyl tRNA during protein synthesis, implying that posttranscriptional modifications in helices 90-92 may be important for ribosome function. RNA fragments were isolated from 23S rRNA by site-directed RNase H digestion. A novel method of mapping modifications by analysis of short, nucleotide-specific, RNase digestion fragments with Matrix Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) was utilized. The MALDI-MS data were complemented by two primer extension techniques using reverse transcriptase. One technique utilizes decreasing concentrations of deoxynucleotide triphosphates to map 2'-O-ribose methylations. In the other, the rRNA is chemically modified, followed by mild alkaline hydrolysis to map pseudouridines (psis). A total of 10 posttranscriptionally methylated nucleotides and 6 psis were detected in the five organisms. Eight of the methylated nucleotides and one psi have not been reported previously. The distribution of modified nucleotides and their locations on the surface of the ribosomal peptidyl transferase cleft suggests functional importance. PMID- 11911367 TI - Selection, design, and characterization of a new potentially therapeutic ribozyme. AB - An in vitro selection was designed to identify RNA-cleaving ribozymes predisposed for function as a drug. The selection scheme required the catalyst to be trans acting with phosphodiesterase activity targeting a fragment of the Kras mRNA under simulated physiological conditions. To increase stabilization against nucleases and to offer the potential for improved functionality, modified sequence space was sampled by transcribing with the following NTPs: 2'-F-ATP, 2' F-UTP, or 2'-F-5-[(N-imidazole-4-acetyl) propylamine]-UTP, 2'-NH2-CTP, and GTP. Active motifs were identified and assessed for their modified NMP and divalent metal dependence. The minimization of the ribozyme's size and the ability to substitute 2'-OMe for 2'-F and 2'-NH2 moieties yielded the motif from these selections most suited for both nuclease stability and therapeutic development. This motif requires only two 2'-NH2-Cs and functions as a 36-mer. Its substrate sequence requirements were determined to be 5'-Y-G-H-3'. Its half-life in human serum is >100 h. In physiologically relevant magnesium concentrations [approximately 1 mM] its kcat = 0.07 min(-1), Km = 70 nM. This report presents a novel nuclease stable ribozyme, designated Zinzyme, possessing optimal activity in simulated physiological conditions and ready for testing in a therapeutic setting. PMID- 11911368 TI - The spliced leader-associated RNA is a trypanosome-specific sn(o) RNA that has the potential to guide pseudouridine formation on the SL RNA. AB - The spliced leader-associated (SLA1) RNA is a trypanosome-specific small RNA with unknown function. SLA1 carries a Sm-like site, and is associated with core Sm proteins. Here we found that SLA1 belongs to a family of hairpin-containing RNAs that are implicated in directing pseudouridylation. A potential for base-pair interaction between SLA1 and spliced leader (SL) RNA agrees with the canonical rules for guiding pseudouridylation on SL RNA. Direct RNA analysis showed that this uridine is indeed pseudouridylated in the SL RNA of Leptomonas collosoma, Leishmania major, and Trypanosoma brucei. This position is conserved in all trypanosomatid SL RNAs. Mutations introduced in the SL RNA to disrupt the interaction domain of SLA1/SL RNA abolished the formation of the pseudouridine. SLA1 is localized both to the nucleolus and nucleoplasm. This study solves a long standing question regarding the function of this novel RNA and describes the first H/ACA RNA, which, unlike all other pseudouridine guides, is also a bona fide snRNA. PMID- 11911369 TI - RNA footprinting analysis using ion pair reverse phase liquid chromatography. AB - Hydroxyl radical footprinting is a powerful technique often employed in characterization of the tertiary interactions between proteins and nucleic acids. Following the generation of a nucleic acid "ladder" either by chemical or enzymatic reactions, the radiolabeled products are traditionally separated by denaturing gel electrophoresis and further quantified by phosphorimaging techniques. Here we report the use of ion pair reverse phase liquid chromatography to analyze the products of an RNA footprinting reaction using fluorescently labeled RNA molecules. This technique offers several advantages over existing procedures, including rapid analysis, automation, and direct quantification of the cleavage products without the need to employ radiolabeling. To illustrate the resolving power of this technique, we have analyzed the products of base hydrolysis, generated from a fluorescently labeled RNA molecule and have subsequently used this method to define the solvent accessibility of the substrate strand as it docks with the hairpin ribozyme. PMID- 11911370 TI - Water counting: quantitating the hydration level of paramagnetic metal ions bound to nucleotides and nucleic acids. AB - Binding of divalent metal ions plays a key role in the structure and function of ribozymes and other RNAs. In turn, the energetics and kinetics of the specific binding process are dominated by the balance between the cost of dehydrating the aqueous ion and the energy gained from inner-sphere interactions with the macromolecule. In this work, we introduce the use of the pulsed EPR technique of 2H Electron Spin-Echo Envelope Modulation (ESEEM) to determine the hydration level of Mn2+ ions bound to nucleotides and nucleic acids. Mn2+ is an excellent structural and functional mimic for Mg2+, the most common divalent ion of physiological interest. Comparison of data in D2O and H2O, with aqueous Mn2+ as a reference standard, allows a robust and precise determination of the number of bound water molecules, and therefore the number of RNA-derived ligands. Examples of applications to the mononucleotide models MnGMP and MnATP, as well as to the paradigmatic RNA system tRNAPhe, are shown. PMID- 11911371 TI - Avian egg size: variation within species and inflexibility within individuals. AB - Egg size is a widely-studied trait and yet the causes and consequences of variation in this trait remain poorly understood. Egg size varies greatly within many avian species, with the largest egg in a population generally being at least 50% bigger, and sometimes twice as large, as the smallest. Generally, approximately 70% of the variation in egg mass is due to variation between rather than within clutches, although there are some cases of extreme intra-clutch egg size variation. Despite the large amount of variation in egg size between females, this trait is highly consistent within individuals between breeding attempts; the repeatability of egg size is generally above 0.6 and tends to be higher than that of clutch size or laying date. Heritability estimates also tend to be much higher for egg size (> 0.5) than for clutch size or laying date (< 0.5). As expected, given the high repeatability and heritability of egg size, supplemental food had no statistically significant effect on this trait in 18 out of 28 (64%) studies. Where dietary supplements do increase egg size, the effect is never more than 13% of the control values and is generally much less. Similarly, ambient temperature during egg formation generally explains less than 15% of the variation in egg size. In short, egg size appears to be a characteristic of individual females, and yet the traits of a female that determine egg size are not clear. Although egg size often increases with female age (17 out of 37 studies), the change in egg size is generally less than 10%. Female mass and size rarely explain more than 20% of the variation in egg size within species. A female's egg size is not consistently related to other aspects of reproductive performance such as clutch size, laying date, or the pair's ability to rear young. Physiological characteristics of the female (e.g. endogenous protein stores, oviduct mass, rate of protein uptake by ovarian follicles) show more promise as potential determinants of egg size. With regards to the consequences of egg-size variation for offspring fitness, egg size is often correlated with offspring mass and size within the first week after hatching, but the evidence for more long-lasting effects on chick growth and survival is equivocal. In other oviparous vertebrates, the magnitude of egg-size variation within populations is often as great or greater than that observed within avian populations. Although there are much fewer estimates of the repeatability of egg size in other taxa, the available evidence suggests that egg size may be more flexible within individuals. Furthermore, in non-avian species (particularly fish and turtles), it is more common for female mass or size to explain a substantial proportion of the variation in egg size. Further research into the physiological basis of egg-size variation is needed to shed light on both the proximate and ultimate causes of intraspecific variation in this trait in birds. PMID- 11911372 TI - Fluctuating asymmetry as an indicator of fitness: can we bridge the gap between studies? AB - There is growing evidence from both experimental and non-experimental studies that fluctuating asymmetry does not consistently index stress or fitness. The widely held--yet poorly substantiated--belief that fluctuating asymmetry can act as a universal measure of developmental stability and predictor of stress mediated changes in fitness, therefore staggers. Yet attempts to understand why the reported relationships between fluctuating asymmetry, stress and fitness are so heterogeneous--i.e. whether the associations are truly weak or non-existent or whether they become confounded during different stages of the analytical pathways remain surprisingly scarce. Hence, we attempt to disentangle these causes, by reviewing the various statistical and conceptual factors that are suspected to confound potential relationships between fluctuating asymmetry, stress and fitness. Two main categories of factors are discerned: those associated with the estimation of developmental stability through fluctuating asymmetry and those associated with the effects of genotype and environment on developmental stability. Next, we describe a series of statistical tools that have recently been developed to help reduce this noise. We argue that the current lack of a theoretical framework that predicts if and when relationships with developmental stability can be expected, urges for further theoretical and empirical research, such as on the genetic architecture of developmental stability in stressed populations. If the underlying developmental mechanisms are better understood, statistical patterns of asymmetry variation may become a biologically meaningful tool. PMID- 11911373 TI - A review of criticisms of phylogenetic nomenclature: is taxonomic freedom the fundamental issue? AB - The proposal to implement a phylogenetic nomenclatural system governed by the PhyloCode), in which taxon names are defined by explicit reference to common descent, has met with strong criticism from some proponents of phylogenetic taxonomy (taxonomy based on the principle of common descent in which only clades and species are recognized). We examine these criticisms and find that some of the perceived problems with phylogenetic nomenclature are based on misconceptions, some are equally true of the current rank-based nomenclatural system, and some will be eliminated by implementation of the PhyloCode. Most of the criticisms are related to an overriding concern that, because the meanings of names are associated with phylogenetic pattern which is subject to change, the adoption of phylogenetic nomenclature will lead to increased instability in the content of taxa. This concern is associated with the fact that, despite the widespread adoption of the view that taxa are historical entities that are conceptualized based on ancestry, many taxonomists also conceptualize taxa based on their content. As a result, critics of phylogenetic nomenclature have argued that taxonomists should be free to emend the content of taxa without constraints imposed by nomenclatural decisions. However, in phylogenetic nomenclature the contents of taxa are determined, not by the taxonomist, but by the combination of the phylogenetic definition of the name and a phylogenetic hypothesis. Because the contents of taxa, once their names are defined, can no longer be freely modified by taxonomists, phylogenetic nomenclature is perceived as limiting taxonomic freedom. We argue that the form of taxonomic freedom inherent to phylogenetic nomenclature is appropriate to phylogenetic taxonomy in which taxa are considered historical entities that are discovered through phylogenetic analysis and are not human constructs. PMID- 11911374 TI - Duet-splitting and the evolution of gibbon songs. AB - Unlike the great apes and most other primates, all species of gibbons are known to produce elaborate, species-specific and sex-specific patterns of vocalisation usually referred to as "songs". In most, but not all, species, mated pairs may characteristically combine their songs in a relatively rigid pattern to produce coordinated duet songs. Previous studies disagree on whether duetting or the absence of duetting represented the primitive condition in gibbons. The present study compares singing behaviour in all gibbon species. Various vocal characteristics were subjected to a phylogenetic analysis using previously published phylogenetic trees of the gibbon radiation as a framework. Variables included the degree of sex-specificity of the vocal repertoire, the occurrence of solo songs, and the preference for a specific time of day for song-production. The results suggest the following scenario for the evolution of gibbon songs: (1) The last common ancestor of recent gibbons produced duet songs. (2) Gibbon duets probably evolved from a song which was common to both sexes and which only later became separated into male-specific and female-specific parts (song-splitting theory). (3) A process tentatively called "duet-splitting" is suggested to have led secondarily from a duetting species to a non-duetting species, in that the contributions of the pair-partners split into temporally segregated solo songs. This appears to be the first time that a non-duetting animal can be shown to be derived from a duetting form. (4) The return to exclusive solo singing may be related to the isolated island distribution of the non-duetting species. PMID- 11911375 TI - Sexual segregation in ungulates: a comparative test of three hypotheses. AB - In most social ungulate species, males are larger than females and the sexes live in separate groups outside the breeding season. It is important for our understanding of the evolution of sociality to find out why sexual segregation is so widespread not only in ungulates but also in other mammals. Sexual body size dimorphism was proposed as a central factor in the evolution of sexual segregation in ungulates. We tested three hypotheses put forward to explain sexual segregation: the predation-risk, the forage-selection, and the activity budget hypothesis. We included in our analyses ungulate species ranging from non dimorphic to extremely dimorphic in body size. We observed oryx, zebra, bighorn sheep and ibex in the field and relied on literature data for 31 additional species. The predation-risk hypothesis predicts that females will use relatively predator-safe habitats, while males are predicted to use habitats with higher predation risk but better food quality. Out of 24 studies on different species of ungulates, females and their offspring chose poorer quality but safer habitat in only eight cases. The forage-selection hypothesis predicts that females would select habitat based on food quality, while males should prefer high forage biomass. In fact, females selected higher quality food in only six out of 18 studies where males and females segregated, in eight studies there was no difference in forage quality and in four studies males were in better quality habitat. The activity budget hypothesis predicts that with increasing dimorphism in body size males and females will increasingly differ in the time spent in different activities. Differences in activity budgets would make it difficult for males and females to stay in mixed-sex groups due to increased costs of synchrony to maintain group cohesion. The predictions of the activity budget hypothesis were confirmed in most cases (22 out of 23 studies). The heavier males were compared to females, the more time females spent foraging compared to males. The bigger the dimorphism in body mass, the more males spent time walking compared to females. Lactating females spent more time foraging than did non-lactating females or males. Whether species were mainly bulk or intermediate feeders did not affect sexual differences in time spent foraging. We conclude that sexual differences in activity budgets are most likely driving sexual segregation and that sexual differences in predation risk or forage selection are additive factors. PMID- 11911376 TI - Some recent advances on the study and understanding of the functional design of the avian lung: morphological and morphometric perspectives. AB - The small highly aerobic avian species have morphometrically superior lungs while the large flightless ones have less well-refined lungs. Two parabronchial systems, i.e. the paleopulmo and neopulmo, occur in the lungs of relatively advanced birds. Although their evolution and development are not clear, understanding their presence is physiologically important particularly since the air- and blood flow patterns in them are different. Geometrically, the bulk air flow in the parabronchial lumen, i.e. in the longitudinal direction, and the flow of deoxygenated blood from the periphery, i.e. in a centripetal direction, are perpendicularly arranged to produce a cross-current relationship. Functionally, the blood capillaries in the avian lung constitute a multicapillary serial arterialization system. The amount of oxygen and carbon dioxide exchanged arises from many modest transactions that occur where air- and blood capillaries interface along the parabronchial lengths, an additive process that greatly enhances the respiratory efficiency. In some species of birds, an epithelial tumescence occurs at the terminal part of the extrapulmonary primary bronchi (EPPB). The swelling narrows the EPPB, conceivably allowing the shunting of inspired air across the openings of the medioventral secondary bronchi, i.e. inspiratory aerodynamic valving. The defence stratagems in the avian lung differ from those of mammals: fewer surface (free) macrophages (SMs) occur, the epithelial cells that line the atria and infundibula are phagocytic, a large population of subepithelial macrophages is present and pulmonary intravascular macrophages exist. This complex defence inventory may explain the paucity of SMs in the avian lung. PMID- 11911377 TI - A cat study. PMID- 11911378 TI - Acculturation, socioeconomic vulnerability, and quality of life in Spanish speaking and bilingual latino HIV-infected men and women. AB - Little is known about the health-related QOL (HRQOL) of low-income, Latino men and women living with HIV Monolingual Spanish-speaking Latino individuals may be at greater risk than bilingual men and womenforpoor HRQOL due to problems associated with language and cultural barriers. This study examined the health status and HRQOL of these two groups of clinic patients. The monolingual group had significantly lower levels of acculturation butdid not differ from the bilingual group on any dimension of health status. This group also reported more disruptions on several dimensions of HRQOL. In a multivariate context, health status variables (as a block) accountedfor the greatest proportion of variance in all three measures of HRQOL. The results did not support the hypothesis that acculturation mediates the impact of health status on HRQOL. More studies are needed to examine the impact of acculturation and socioeconomic vulnerability on health outcomes. PMID- 11911379 TI - The decision to take hormone replacement therapy among women with disabilities. AB - Whereas making decisions during menopause can be challenging for all women, those with physical impairments face special issues with respect to menopause in general and hormone replacement therapy (HRT) in particular. In this correlational study the authors explored the factors such women consider when making decisions about HRT One hundred sixty-seven women with physical impairments throughout the United States completed surveys concerning their attitudes and knowledge about HRT Approximately half the menopausal women were currently taking HRT The strongest predictor of HRT use was women's perceptions of their health care providers opinions about their taking HRT, combined with their motivation to comply with the provider's recommendation. Thisfinding points to the significant role that nurses and other health care providers play in assisting women to make informed health care choices during menopause. PMID- 11911380 TI - Examination of the transtheoretical model in current smokers. AB - Although the transtheoretical model of behavior change has frequently been used as a basisfor smoking-cessation programs, in very few studies have the study variables been linked with the theoretical concepts. This study used a convenience sample of 79 current smokers to determine the relationships among the stages of change and the processes of change. Logistic regressions were used in order to determine whether certain processes were related to specific stages and whether specific processes were related to movement among the stages. Participants in the precontemplation stage and the preparation stage were found to rely on specific processes, whereas those in the contemplation stage did not. PMID- 11911382 TI - Using hierarchical cluster analysis in nursing research. AB - This article is a pedagogical piece on hierarchical cluster analysis, a method for investigating the structure underlying data. Such methods are useful for finding similar groups of cases in data sets when it is not known a priori how many groups are present. The article is laid out asfollows: First, a brief history and overview of the methods is presented; second, an illustrative example with a small hypothetical data set is used to clarify fundamental concepts; third, hierarchical cluster analysis is applied to a data set from the author's own program of research to illustrate one way in which the methods may be employed in nursing research; fourth, the limitations of the methods are discussed; and finally, a list of suggested readings, at varying levels of detail. are provided for the interested researcher. PMID- 11911381 TI - Help-seeking and social support in Japanese sojourners. AB - Research shows that social support is essential for healthy psychological functioning. Help seeking and social support are soc ial processes shaped by cultural understandings about how need should be expressed, to whom, and in what circumstances. This study used grounded theory methodology to examine how cultural factors regulate help seeking and social support in a sample of 25 Japanese sojourners' wives living in America. Culturally based social edicts such as mutual responsibility and in-group solidarity were found to promote help seeking and social support. In contrast, culturally specific factors such as enryo (polite deference) hierarchy, and the cultural rules governing reciprocity inhibited these behaviors. From these data, a cultural model of social exchange allowing for cultural diversity, is proposed. This model can increase the effectiveness of nursing interventions aimed at community-based health promotion. PMID- 11911383 TI - Comparative judgment of numerosity and numerical magnitude: attention preempts automaticity. AB - It is commonly believed that humans are unable to ignore the meanings of numerical symbols, even when these meanings are irrelevant to the task at hand. In 5 experiments, the authors tested the notion of automatic activation of numerical magnitude by asking participants to compare, while timed, pairs of numerical arrays on either numerosity or numerical value. Garner and Stroop effects were used to gauge the degree of interactive processing. The results showed that both effects were sensitive to the discriminability of values along the constituent dimensions, to the number of stimulus values used, and to practice and motivation. Notably, Stroop and Garner effects were eliminated under several conditions. These findings are incompatible with claims of obligatory activation of meaning in numerical processing, and they cast doubt on theories positing automatic processing of semantic information for alphanumerical symbols. PMID- 11911384 TI - Comparing prototype-based and exemplar-based accounts of category learning and attentional allocation. AB - Exemplar theory was motivated by research that often used D. L. Medin and M. M. Schaffer's (1978) 5/4 stimulus set. The exemplar model has seemed to fit categorization data from this stimulus set better than a prototype model can. Moreover, the exemplar model alone predicts a qualitative aspect of performance that participants sometimes show. In 2 experiments, the authors reexamined these findings. In both experiments, a prototype model fit participants' performance profiles better than an exemplar model did when comparable prototype and exemplar models were used. Moreover, even when participants showed the qualitative aspect of performance, the exemplar model explained it by making implausible assumptions about human attention and effort in categorization tasks. An independent assay of participants' attentional strategies suggested that the description the exemplar model offers in such cases is incorrect. A review of 30 uses of the 5/4 stimulus set in the literature reinforces this suggestion. PMID- 11911385 TI - Visual marking: dissociating effects of new and old set size. AB - Visual marking makes it possible to ignore old items during search. In a typical study, old items are previewed 1 s before adding an equal number of new items, one of which is the target. Previewing half of the items reduces the search slope relating response time (RT) to overall set size by half. However, this manipulation sometimes only reduces overall RT but not search slope (Experiment 1). By orthogonally varying the numbers of old and new items, Experiment 2 shows that old and new set sizes interactively affect visual marking. Given a constant new set size, the size of the old set has negligible effect on RT. However, increasing the new set size reduces the preview benefit in overall RT. Experiment 3 shows that this reduction may be restricted to paradigms that use temporal segregation cues. Studies should vary old and new set size orthogonally to avoid missing a visual marking effect where one may be present. PMID- 11911386 TI - The effect of negation on deductive inferences. AB - Research shows that negation can suppress the activation of propositions presented explicitly in text, but does negation have a similar effect on propositions that can be inferred? That is, does negation inhibit the inference process? Four experiments investigated whether a deductive inference that produces a negated conclusion (therefore not a) is made as readily as a similar inference form that yields an affirmative conclusion (therefore a). A combination of naming latencies, verification times, and reading times indicate that negation does not affect the deductive inference process itself, although it may inhibit the activation of inferred concepts. PMID- 11911387 TI - The effects of surface and structural feature matches on the access of story analogs. AB - Competing theories of analogical reasoning have disagreed on the relative contributions of surface and structural features to the access of previously read base stories when one is reading a current cue story. A key limitation of the prior work was that surface and structural feature overlap between bases and cues was not manipulated precisely. The present study systematically manipulated the number of surface and structural matches to determine their relative effect on access. Results involving reminding and reading-time measures suggest that surface and lower-order structural features affected access about equally, at least when a higher-order relation (HOR) was shared between a base and cue story. When a HOR was not shared, surface feature overlap continued to affect access while lower-order structural features had a less reliable effect. Models of access might need to be adjusted to account for these phenomena. PMID- 11911388 TI - Spatial updating of locations specified by 3-d sound and spatial language. AB - Blind and blindfolded sighted observers were presented with auditory stimuli specifying target locations. The stimulus was either sound from a loudspeaker or spatial language (e.g., "2 o'clock, 16 ft"). On each trial, an observer attempted to walk to the target location along a direct or indirect path. The ability to mentally keep track of the target location without concurrent perceptual information about it (spatial updating) was assessed in terms of the separation between the stopping points for the 2 paths. Updating performance was very nearly the same for the 2 modalities, indicating that once an internal representation of a location has been determined, subsequent updating performance is nearly independent of the modality used to specify the representation. PMID- 11911389 TI - Feature modulation search: a novel memory search model that extends the perceptual interference effect to musical stimuli. AB - Although most research in the perceptual interference literature has used visual stimuli, recent experiments have replicated this effect using musical stimuli. Two experiments investigated whether musical and visual demonstrations of the effect are produced by a common mechanism by examining whether the provision of incorrect cues would impair melodic object identification in the perceptual interference paradigm. Whereas incorrect cues typically do not affect visual object identification, they reduced melody identification both when the subjects generated the cues and when the experimenter provided them. A novel model (feature modulation search) is presented to account for perceptual interference in musical stimuli. The model has potential applications for the larger perceptual interference literature and for related phenomena in which target related information impairs cognition. PMID- 11911390 TI - Outstanding memories: the positive and negative effects of nudes on memory. AB - A picture of a nude isolated in a series of pictures of clothed models was quite memorable, but when compared with a clothed picture, the nude impaired memory for background picture derails as well as pictures immediately following the nude (anterograde amnesia). Recall of details given picture-gist recall, as well as recognition of person details, were equivalent in the clothed and nude conditions. A clothed picture isolated in a series of nudes did not hurt memory for background information, nor did it produce anterograde amnesia. Apparently, distinctiveness supports good memory for the gist of the nude or clothed pictures. However, distinctiveness is not responsible for the anterograde amnesia and poor memory for background details found with nudes. PMID- 11911391 TI - Gender is a dimension of face recognition. AB - In an experiment, the authors investigated the impact of gender categorization on face recognition. Participants were familiarized with composite androgynous faces labeled with either a woman's first name (Mary) or a man's first name (John). The results indicated that participants more quickly eliminated faces of the opposite gender than faces of the same gender than the face they were looking for. This gender effect did not result from greater similarity between faces of the same gender. Rather, early gender categorization of a face during face recognition appears to speed up the comparison process between the perceptual input and the facial representation. Implications for face recognition models are discussed. PMID- 11911393 TI - Conjunction error rates on a continuous recognition memory test: little evidence for recollection. AB - Two experiments examined conjunction memory errors on a continuous recognition task where the lag between parent words (e.g., blackmail, jailbird) and later conjunction lures (blackbird) was manipulated. In Experiment 1, contrary to expectations, the conjunction error rate was highest at the shortest lag (1 word) and decreased as the lag increased. In Experiment 2 the conjunction error rate increased significantly from a 0- to a 1-word lag, then decreased slightly from a 1- to a 5-word lag. The results provide mixed support for simple familiarity and dual-process accounts of recognition. Paradoxically, searching for an item in memory does not appear to be a good encoding task. PMID- 11911392 TI - Brain-potential evidence for the time course of access to biographical facts and names of familiar persons. AB - On seeing familiar persons, biographical (semantic) information is typically retrieved faster and more accurately than name information. Serial stage models explain this pattern by suggesting that access to the name follows the retrieval of semantic information. In contrast, interactive activation and competition (IAC) models hold that both processes start together but name retrieval is slower because of structural peculiarities. With a 2-choice go/no-go procedure based on a semantic and a name-related classification, the authors tested differential predictions of the 2 alternative models for reaction times (RTs) and lateralized readiness potentials (LRP). Both LRP (Experiment 1) and RT (Experiment 2) results are in line with IAC models of face identification and naming. PMID- 11911394 TI - On the form of ROCs constructed from confidence ratings. AB - A classical question for memory researchers is whether memories vary in an all-or nothing, discrete manner (e.g., stored vs. not stored, recalled vs. not recalled), or whether they vary along a continuous dimension (e.g., strength, similarity, or familiarity). For yes-no classification tasks, continuous- and discrete-state models predict nonlinear and linear receiver operating characteristics (ROCs), respectively (D. M. Green & J. A. Swets, 1966; N. A. Macmillan & C. D. Creelman, 1991). Recently, several authors have assumed that these predictions are generalizable to confidence ratings tasks (J. Qin, C. L. Raye, M. K. Johnson, & K. J. Mitchell, 2001; S. D. Slotnick, S. A. Klein, C. S. Dodson, & A. P. Shimamura, 2000, and A. P. Yonelinas, 1999). This assumption is shown to be unwarranted by showing that discrete-state ratings models predict both linear and nonlinear ROCs. The critical factor determining the form of the discrete-state ROC is the response strategy adopted by the classifier. PMID- 11911395 TI - The absence of a gender congruency effect in romance languages: a matter of stimulus onset asynchrony? AB - Using the picture-word interference paradigm, H. Schriefers and E. Teruel (2000) found that in German the grammatical gender of the distractor word affects the production of phrases composed of article+picture name: Latencies were longer for picture-word pairs of different genders. However, the effect was found only at positive stimulus onset asynchronies (SOAs; i.e., when pictures were presented 75 or 150 ms earlier than word distractors). This gender congruency effect is not obtained in Romance languages. The present article examines whether in these languages, as in German, the effect appears at positive SOAs. No effect was observed in Italian and Spanish at positive SOAs. An account is proposed to explain why the gender congruency effect is obtained in Germanic (Dutch and German) but not in Romance languages. PMID- 11911396 TI - Why aggregated learning follows the power law of practice when individual learning does not: comment on Rickard (1997, 1999), Delaney et al. (1998), and Palmeri (1999). AB - The power law of practice is often considered a benchmark test for theories of cognitive skill acquisition. Recently, P. F. Delaney, L. M. Reder, J. J. Staszewski, and F. E. Ritter (1998), T. J. Palmeri (1999), and T. C. Rickard (1997, 1999) have challenged its validity by showing that empirical data can systematically deviate from power-function fits. The main purpose of the present article is to extend their explanations in two ways. First, the authors empirically show that abrupt changes in performance are not necessarily based on a shift from algorithm to memory-based processing, but rather and more generally, that they occur whenever a more efficient task strategy is generated. Second, the authors show mathematically and per simulation that power functions can perfectly fit aggregated learning curves even when all underlying individual curves are discontinuous. Therefore, the authors question conclusions drawn from fits to aggregated data. PMID- 11911397 TI - T-cell associated antigen-positive B-cell lymphoma. AB - We immunophenotyped 128 patients with B-cell non-Hodgkin's lymphoma (B-NHL) of various histological subtypes using two-color flow cytometry (FCM), and found that lymphoma cells obtained from 31 patients (24.2%) coexpressed at least one of the following T-cell associated antigens (T-Ag); CD2 (2.3%), CD5 (18.0%) or CD7 (6.3%). Moreover, 3 patients expressed two kinds of T-Ag (CD2/CD5, CD2/CD7 or CD5/CD7) as reported by other investigators. Though we could not find coexpression of CD3, CD4 or CD8 antigen in any patients analyzed in our study, such T-Ag(+) B-NHL have also been reported in the literature. As clinical features, extranodular involvement and higher International Prognostic Index (high and high intermediate) seemed more frequent in T-Ag(+) B-NHL than T-Ag(-) B NHL in our study. Such prognostic significance of T-Ag expression is also reported by other investigators especially in CD5(+) diffuse large B-cell lymphoma. In addition, two-color FCM for detecting such aberrant T-Ag expression in B-NHL is useful for monitoring the minimal residual disease in the subgroup with T-Ag(+) B-NHL. PMID- 11911398 TI - Chromosomal DNA and p53 stability, ubiquitin system and apoptosis in B-CLL lymphocytes. AB - The ubiquitin system regulates diverse biological processes such as DNA replication and repair, biogenesis of ribosome, peroxisome and nucleosome, cell cycle, stress response and signal transduction pathways. Thus, the reported role of the ubiquitin system in apoptotic death control as well the alteration of its control in carcinogenesis should come as no surprise. Indeed, we and other groups have reported that the ubiquitin system is involved in apoptotic cell death of normal human lymphocytes and that this control is altered in B lymphocytes derived from chronic lymphocytic leukemia patients (B-CLL), rendering these malignant cells hypersensitive to specific inhibition of protein degradation/processing through proteasomal function. This approach recently allowed us to demonstrate that the stability of the tumor suppressor and pro apoptotic protein p53 is differentially regulated in B-CLL versus normal lymphocytes and that this difference might at least partly explain the impaired response of B-CLL lymphocytes to apoptotic death activation. These results strongly suggest an imbalance in p53 regulation in B-CLL cells that leads to a variable response to DNA damage and constitutively expressed chromosomal instability. The question we and others would like to address is whether this alteration, or more likely a subset of alterations of the ubiquitin-proteasome pathway, is specific to B-CLL malignancy or if it is a hallmark of cancer cells in general. In either case, a better understanding of the ubiquitin-dependent control of apoptosis should pave the way towards a methodological approach for in vitro development of discriminating treatments which may be of potential usefulness in clinical trials of B-CLL. PMID- 11911400 TI - CD105 (endoglin) expression on hematopoietic stem/progenitor cells. AB - Endoglin (CD105) is a component of the transforming growth factor-beta (TGF-beta) receptor (TGF-betaR) complex. Together with betaglycan, CD105 is considered as a TGF-betaR accessory molecule (also called TGF-betaRIII), but its functions in the receptor-ligand interactions are still poorly understood. A small subset of human CD34+ hematopoietic stem/progenitor cells that has phenotypic and functional features suggestive of very primitive hematopoietic cells expresses the CD105 antigen. CD34+/CD105+ cells recirculate in the peripheral blood of mobilized subjects and can be purified by immunomagnetic isolation strategies. The hematopoietic potential of these CD34+/CD105+ cells appears to be sustained by a combination of hematopoietic and non-hematopoietic cytokines, which comprises Flt3 ligand, erythropoietin, interleukin-15 and vascular endothelial growth factor. Endogenous TGF-beta1 is a crucial factor for the maintenance of CD34+/CD105+ immaturity acting through positive modulation of both CD105 and CD34 molecules in the absence of relevant effects on the cell cycle profile. CD105 is absent on very primitive CD34-/lineage-/CD45+ (CD34-Lin-) human hematopoietic cells isolated from cord blood. However, in vitro exposure of CD34-Lin- cells to exogenous TGF-beta1 causes the appearance of a discrete population of CD34+/CD105+ cells. Collectively, available data on CD105 expression and function in primitive hematopoiesis indicate that this molecule could cooperate with the dissociation of TGF-beta1 cell cycle effects from its other effects on cell survival and differentiation. PMID- 11911399 TI - Novel insights into the mechanism of t(14;18)(q32;q21) translocation in follicular lymphoma. AB - The t(14:18)(q32:q21) chromosomal translocation and the ensuing overexpression of the BCL-2 proto-oncogene are strongly associated with the pathogenesis of follicular lymphoma. At the molecular level, the translocation process arises from the illegitimate rearrangement between the BCL-2 proto-oncogene and the immunoglobulin heavy chain (IgH) locus. Due to the presence of the D(H) and J(H) gene segments from the IgH locus as well as de novo nucleotide additions at the breakpoints, the translocation process has been assumed to result from a mistake occurring during V(D)J recombination in early B-cells in the bone marrow. However, recent detailed molecular analyses of both the direct and reciprocal breakpoints have revealed that the t(14;18) translocation is a more complex process than previously thought, and have challenged this traditional view. Here we review these observations, and discuss the intriguing possibility that t(14;18) translocation could preferentially occur in the germinal centers during receptor revision, and involves both V(D)J recombination and somatic hypermutation mechanisms. PMID- 11911401 TI - Transplantation of autologous peripheral blood progenitor cells: impact of CD34 cell selection on immunological reconstitution. AB - Peripheral blood progenitor cells (PBPC) represent an ideal source of stem cells for autologous transplantation because of technical advantages and more favourable engraftment kinetics. The reconstituion of a functional immune system occurs earlier in patients transplanted with cytokine-mobilized autologous PBPC compared with bone marrow; because of the greater T-cell content in PBPC products, donor-derived antigen-specific T-cells transferred with the graft might contribute to short-term immunity in transplant recipients. Despite a prompt reconstitution of B- and T-cell numbers, both B- and T-cell function are profoundly impaired for a prolonged period of time after PBPC infusion. The positive selection of CD34+ cells might provide effective tumor cell purging without compromising hematopoietic recovery in patients with acute leukemia, multiple myeloma, breast cancer and non-Hodgkin's lymphoma, whose autografts have been reported to contain malignant cells which might promote disease relapse. However, the incidence of viral infections in the early posttransplant period might be increased after CD34-selected compared with unmanipulated PBPC transplants, as a result of the lack of accessory and immune cells in the graft. The purpose of this review is to provide an update on immunological reconstitution after transplantation of autologous PBPC; in particular, emphasis will be placed on the mechanisms of immune dysfunction after the infusion of unmanipulated and CD34-selected autografts. PMID- 11911402 TI - Increased chemokine receptor expression and the infiltration of lymphoid organs. AB - Lymphocyte passage across endothelium into lymph nodes and Peyer's patches is a multistep process that involves selectin-supported rolling, followed by a triggering event, and then firm integrin-mediated adhesion. The combined actions of adhesion molecules and chemokines largely determine the migratory behavior of lymphocytes. Chemokines are known to mediate their effects through 7 transmembrane-domain G-protein-coupled receptors. Here, we review the role of chemokines and their receptors in lymphocyte's migration into lymphoid organs. PMID- 11911403 TI - Myelodysplastic syndrome and pregnancy: the Mayo Clinic experience. AB - Although the myelodysplastic syndrome (MDS) is most common in the elderly, younger patients, including women of child-bearing age, may be affected. The association of MDS with pregnancy appears to be very rare: fewer than 25 cases have been reported. We report the outcomes of seven pregnancies in four women seen at the Mayo clinic between 1983 and 2000. Three of the women were found to have MDS when an abnormal complete blood count was detected during routine prenatal care. The fourth patient had an apparently congenital MDS, and suffered three spontaneous abortions before undergoing premature menopause as a result of pelvic irradiation for vulvar cancer. We discuss some of the unique concerns regarding pregnant patients with MDS. As women give birth at older ages and as more young persons survive cancer, MDS and pregnancy are likely to be seen together with increasing frequency. PMID- 11911404 TI - Efficacy of splenectomy for patients with mantle cell non-Hodgkin's lymphoma. AB - The purpose of this study was to define the role of splenectomy in patients (pts) with mantle cell lymphoma (MCL) with regard to improving cytopenias and symptoms of splenomegaly. 26 pts with MCL underwent splenectomy between January 1987 and October 1999 and were followed prospectively for hematologic response and operative morbidity and mortality. A positive response was defined at 1 month of follow-up as: a hemoglobin of > or = 1.0 g/dl in a pt with a preoperative value < 11.0 g/dl; or a platelet count of > or = 100 x 10(9)/L in a pt with a preoperative value < 100 x 10(9)/L. A positive hematologic response was achieved in 69.2% of pts with preoperative anemia, 90% with thrombocytopenia, and 50% with both anemia and thrombocytopenia. The peri- and post-operative morbidity were 3.8 and 19.2%, respectively, the operative mortality was 0%. The median duration of hospitalization was six days. Four (15.4%) pts have not required chemotherapy after splenectomy. Three of these four were previously untreated and they have maintained stable disease for eight years after splenectomy without chemotherapy. Eight additional pts did not require chemotherapy for > 13 months after splenectomy. These results suggest that splenectomy may provide durable remission in selected pts with refractory cytopenias or symptoms related to splenomegaly in pts with MCL. There is a subset of pts that have prolonged disease stabilization without the requirement for immediate chemotherapy after splenectomy. PMID- 11911405 TI - Hydroxyurea-associated platelet count oscillations in polycythemia vera: a report of four new cases and a review. AB - Cyclic oscillations in the peripheral blood platelet count were recently described in two patients with polycythemia vera (PV) receiving therapy with hydroxyurea (HU). This phenomenon can make proper HU dosing very challenging and may be especially problematic in PV patients who are at risk for thrombohemorrhagic complications. In this report, we describe four new cases of HU-associated platelet oscillations in patients with PV and extend our observations on one of the two previously reported cases. We also review cyclic thrombocytosis within the broader context of periodic hematopoiesis. A thrombopoietin-mediated negative feedback loop is central to most models of periodic thrombopoiesis, but this is probably an oversimplification. It is possible that HU destabilizes the delicate balance of thrombopoietin/c-Mpl signaling in megakaryocytic lineage hematopoiesis that is already markedly altered in PV; this hypothesis remains speculative. For patients who develop oscillatory variation in their platelet counts associated with HU use, keeping the HU dose constant may result in damping or termination of the cycles. However, this strategy is not always successful. PMID- 11911406 TI - High dose cyclophosphamide plus recombinant human granulocyte-colony stimulating factor (rhG-CSF) in the treatment of follicular, low grade non-Hodgkin's lymphoma: CALGB 9150. AB - The main objectives of this study were to determine the feasibility of administering high doses of cyclophosphamide plus recombinant human granulocyte colony stimulating factor (rhG-CSF) every 14-21 days to patients with follicular small cleaved cell lymphoma. For each patient, the treatment was not considered feasible if fewer than four cycles of cyclophosphamide chemotherapy could be administered on schedule (i.e. at least every 29 days) or (1) hospitalization of the patient for longer than three days was necessary for neutropenic fever (38 degrees C) or bacteriologically documented infection in > 50% of the cycles, or (2) grade > or = 2 hemorrhage in association with thrombocytopenia of grade > or = 3 severity occurred in > 50% of the cycles or (3) non-hematologic toxicity (excluding nausea/vomiting and alopecia) of grade > or = 3 occurred in > 50% of cycles. The goal was to have a treatment program feasible in 75% or more of the treated patients. The secondary objectives were to determine the toxicities, the complete and partial response rates, and the time to treatment failure (TTF). The trial also attempted to assess the effectiveness of this treatment program in eradicating Bcl-2 rearrangements by PCR, and to assess complete remission duration in relationship to PCR results in patients who respond to this chemotherapy program. Patients were required to have histologically documented non-Hodgkin's lymphoma of the subtypes follicular, predominantly small cleaved cell (IWF-B) or follicular mixed, (IWF-C). Patients were required to have Stage IV disease including histologic evidence of bone marrow involvement. Measurable disease was required and patients were also required to have one of the following risk factors: > or = 2 extranodal sites, node or nodal group > or = 5 cm. Submission of fresh bone marrow for molecular genetic studies for the presence of Bcl-2-Ig fusion DNA was mandatory in previously untreated patients. Patients had to be between 18 and physiologic age 55 years (carefully selected patients over age 55 years were also eligible), expected survival > 2 years, performance status 0-1, and have adequate renal, hepatic and bone marrow function, and a cardiac ejection fraction > or = 50%. Cyclophosphamide 4.5 g/m2 i.v. was given with mesna every 14 days with rhG-CSF support. Twenty-nine patients were accrued to this trial. The median follow-up time is 5.0 years, with a range of 2.5-6.7 years. The overall response rate was 75% (9 CRs 37.5%, 9PRs 37.5%). The median duration of survival is 5.53 years. The 1-year estimated probability of freedom from treatment failure was 50% and of survival at 1 year was 92%. No strong association was observed between TTF and age, symptomatic stage, histology performance status, number of extranodal sites or baseline Bcl-2 status. At 3 years the survival of all patients was 78% and failure free survival was 17%. 15 (62%) of the 24 eligible previously untreated patients met the criteria for feasibility specified in the protocol. The 95% CI for the feasibility rate is (44 and 82%). Twenty-two of the 24 (92%) previously untreated patients had specimens submitted for testing for Bcl-2 rearrangements. Thirteen of the 22 (59%) were found to have rearrangements at baseline. Post-treatment specimens were submitted for seven of the 13 patients. Four of the seven converted to Bcl-2 negative following treatment. Eight of 13 Bcl-2 positive patients (62%) had a clinical response to treatment. The 95% exact binomial CI for the total response rate in this subgroup is (28 and 88%). This study demonstrates that repetitive doses of cyclophosphamide at 4.5 g/m2 every two weeks with rhG-CSF support can be administered to selected younger patients with advanced follicular lymphoma with morphologic involvement of the bone marrow with acceptable non-hematologic toxicity. PMID- 11911407 TI - Treatment of acute promyelocytic leukemia with arsenic trioxide: clinical and basic studies. AB - Arsenic trioxide (As2O3) has recently been identified as an effective drug in the treatment of newly diagnosed and relapsed acute promyelocytic leukemia (APL) without cross-resistance to all-trans retinoic acid and achieved complete remission rates of 80-90% according to most reports. With intravenous infusion at a dose of 0.08-0.16 mg/kg daily, a course of 28-42 days is required to induce remission. As2O3 in combination with chemotherapy as postremission therapy results in longer survival than arsenic alone. In vitro, As2O3 exerts dose dependent dual effect; triggering apoptosis at relatively high concentration (0.5 2.0 micromol/l), which is associated with the disruption of mitochondrial transmembrane potentials, while inducing partial differentiation at low concentration (0.1-0.5 micromol/l), which might be related to retinoic acid signaling pathway. Importantly, at both concentrations, As2O3 can degrade PML (promyelocytic leukemia) -RAR alpha (retinoic acid receptor), an oncoprotein that has a central role in leukemogenesis. PMID- 11911408 TI - Second malignancies after treatment for Hodgkin's disease. AB - The occurrence of treatment-related second malignancy following Hodgkin's disease (HD) has now been recognized as a major problem. The purpose of this study was to review our experience with second malignancies in patients treated for Hodgkin's disease, comparing the results with the international literature data. Six hundred and sixty five patients with HD were treated in our department, between 1978 and 1996. Second neoplasm developed in 32 cases (4.8%). Seven secondary hematological malignancies were observed: four acute nonlymphocytic leukemias, two non-Hodgkin's lymphomas and one chronic myeloid leukemia. Among patients with second hematological malignancies, the mean age at diagnosis of HD was 44 years and the mean interval until the development of second malignancy was 6.1 years. Five patients received chemo- and radiotherapy and in two cases chemotherapy was used. Three of the seven patients are alive. Twenty-five patients have had solid tumors, affecting lung (5), breast (3), colon (3), stomach (2), urinary bladder (2), head-and-neck (1), thyroid gland (1), esophagus (1), liver (1), pancreas (1), furthermore, three sarcomas and two malignant melanomas were observed. Their mean age at the diagnosis of HD was 46 years and the mean period of latency was 8.3 years. Chemotherapy was applied to nine patients, 16 patients received both chemo- and radiotherapy. Eleven patients had solid tumors in the region irradiated earlier. Ten out of the 25 patients are alive, three patients' present state is unknown. Since alkylating agents increase the risk of leukemia and irradiation contributes mainly to other malignancies, future treatment protocols should attempt to reduce the most serious consequence of therapy without compromising the survival. It is necessary to investigate the impact of additional risk factors. Careful, lifelong observation is indicated for patients with HD, with special attention given to new clinical signs and symptoms. PMID- 11911409 TI - Dose escalation of ara-c may improve response rates in a subgroup of chronic myeloid leukemia patients with poor response to interferon-alpha and low-dose ara C. AB - The present analysis was performed to evaluate the impact of cytosine arabinoside (ara-C) dose escalation on hematological and cytogenetic responses in patients with chronic myelogenous leukemia (CML) who failed to respond to low-dose ara-C (LD ara-C) at a dose of 10 mg/m2/d over 10 days per month and interferon-alpha (IFNalpha, 3.5 MU/d). Following the same administration schedule, dose escalation of ara-C to 15 and 20 mg/m2/d 1-10 was performed in 36 of 119 patients (30%) due to inadequate hematological response and/or disease progression. As a result, improvement of hematological and cytogenetic responses was achieved in 22 (61%) and nine (25%) patients, respectively. Escalated ara-C dose levels were usually well tolerated, although some patients experienced deterioration of preexisting side effects. Our results support the critical role of ara-C dose towards a better disease control in CML. PMID- 11911410 TI - Risk group definition in children with acute myeloid leukemia by calculating individual risk factors on the basis of a multivariate stepwise Cox regression analysis. AB - To define risk groups in children with acute myeloid leukaemia (AML), we conducted a multivariate stepwise Cox regression analysis of three consecutive multicentre studies in East Germany. The total number of patients was 240, but cytogenetics and remission status on day 15 were routinely investigated only in the most recent study, AML-III/93 (78 patients). We derived an equation to calculate individual risk factors, determined those risk factors for all patients of study AML-III/93 and divided them into three groups with 26 patients in each. The variables in the equation were: WBC, FAB-type, auer rods, cytogenetics and response status on day 15. The event-free survival was 80% in the low risk, 55% in the intermediate risk and 15% in the high risk group. Our results strongly suggest that calculating individual risk factors on the basis of a multivariate stepwise Cox regression analysis is a useful tool in defining risk groups. PMID- 11911412 TI - Multiple bone lesions after allogeneic bone marrow transplantation in a patient with relapsed adult acute lymphoblastic leukemia: minimal residual disease analysis may predict extramedullary relapse. AB - We describe a patient with acute lymphoblastic leukemia (ALL, L2) who relapsed with multiple bone lesions after allogeneic bone marrow transplantation (allo BMT). Allo-BMT was performed from an HLA-identical sibling during the first hematological complete remission (CR). Minimal residual disease (MRD) assessed by polymerase chain reaction (PCR) with primers for T cell receptor delta (TCRdelta) gene became positive in the bone marrow sample on day 46 after allo-BMT. On day 113, the patient complained of a painful tumor at the right clavicle. The examination of biopsy specimen revealed infiltration of leukemic cells. After partial response was achieved by local radiotherapy, disseminated bone lesions were demonstrated by 99mTC scintigraphy scan, followed by bone marrow relapse on day 137. The patient died of cardiac tamponade on day 236 after Allo-BMT. MRD assessed by PCR assay for TCRdelta gene in the bone marrow is useful for the prediction of extramedullary as well as medullary relapse after BMT. PMID- 11911411 TI - High-dose chemotherapy with hematopoietic stem cell transplantation is effective for nasal and nasal-type CD56+ natural killer cell lymphomas. AB - CD56+ natural killer (NK) cell lymphomas occur frequently in the nasal and nasopharyngeal regions and carry a poor prognosis. We have studied seven cases with NK-cell lymphomas. These lymphomas showed the following immunophenotype: CD56+, CD2+, sCD3- and Epstein-Barr virus-encoded small RNAs (EBERs)+. Six patients had localized (stage I or II) disease involving the nasopharyngeal region, while one had stage III disease. One patient with stage I disease achieved a complete remission (CR) after treatment with involved-field irradiation, but subsequently relapsed and died. The remaining six patients received combination chemotherapy as primary treatment: five patients with localized stage I or II disease and one patient with advanced stage III disease. Responses to initial chemotherapy were generally poor. These six patients received a variety of salvage chemotherapy regimens, but never achieved a CR. Subsequently, four of six patients showed a highly aggressive clinical course and died of disseminated disease within 1 year from the diagnosis. Three of six patients received high-dose chemotherapy supported by syngeneic, autologous or allogeneic peripheral blood stem cell transplantation. Two of the three transplant patients achieved a CR and are now surviving in continuous CR. Our clinical experience suggests that myeloablative high-dose chemotherapy and bone marrow rescue by hematopoietic stem cell transplantation may be an effective salvage treatment modality for refractory NK-cell lymphomas and could be considered as a part of the initial therapy for these patients. PMID- 11911413 TI - Widespread bone disease in acute myeloid leukaemia. AB - We describe a 11-year-old boy with acute myeloid leukaemia who presented with widespread bone disease. Spine X-rays revealed multiple crush fractures and there were multiple hot spots on the bone scan. The bone-mineral density was markedly reduced but there was no hypercalcaemia or hypercalcuria. Bone marrow aspirate revealed 98% blast cells and a balanced translocation between chromosomes 10 and 17 in seven of nine metaphases. Plasma interleukin-6 level before chemotherapy was high at 53 pg/ml. We postulate that the mechanism for bony destruction in this case was similar to that in the adult disease myeloma. PMID- 11911414 TI - Association of CD38 antigen expression with other prognostic parameters in early stages of chronic lymphocytic leukemia. AB - The expression of the surface molecule CD38 on B cell chronic lymphocytic leukemia (B-CLL) cells has recently been described as a prognostic marker for patient survival. We have analyzed CD19/CD38 expression in 81 patients with predominantly early stages of B-CLL (69 Binet A, seven Binet B, five Binet C). Sixty-two patients (77%) had less than 30% CD38+/CD19+ cells, while 19 (23%) had > or = 30%. There was a significant association between Binet stages (A vs. B+C, p < 0.0001), Rai stages (0-II vs. III+IV, p < 0.001) and CD38 expression, confirming the published cut-off level of 30%. A particularly strong association between CD38 expression was found with soluble CD23 (sCD23) levels of > or = 2000 U/ml (p < 0.0001) and beta2-microglobulin (beta2 MG) serum levels of > or = 3 mg/l (p < 0.0001) indicating that CD38 is a marker of tumor mass as well as disease progression. A borderline association was found with lymphocyte doubling time (LDT) < 12 months (p = 0.05) due to low patient numbers, while there was no association with age, sex or immunoglobulin deficiency. Discordant results were obtained in a number of patients: 10 of 69 patients (14%) with Binet A had a CD38 > or = 30% while three of seven patients with Binet B had a CD38 < 30%. In these two subgroups CD38 and other prognostic factors gave discrepant results. Due to the early stage and short median observation time (12 months. range 1-24 months), calculations concerning patient survival were not performed. However, our data show a strong association between CD38 and other known prognostic factors. The results also suggest that this factor is not always reliable in Binet A patients. PMID- 11911415 TI - Tamoxifen-based treatment induces clinically meaningful responses in multiple myeloma patients with relapsing disease after autotransplantation. AB - Tamoxifen has been shown to induce apoptosis in multiple myeloma cell lines and primary myeloma cells. Daily tamoxifen was given to 12 consecutive multiple myeloma patients who either relapsed following autologous stem cell transplantation (11) or had progressive disease on conventional chemotherapy (1). Methotrexate was also given biweekly to enhance the antiangiogenetic effect. Two patients achieved complete remission lasting 8 and 18 months. Two additional patients had stable disease (SD) for 7 and 11 months. All responders were men and the earliest signs of response were seen after approximately 6-8 weeks of treatment. The regimen was very well tolerated. Speculations about a possible mechanism of action of tamoxifen in multiple myeloma are discussed. PMID- 11911416 TI - Rituximab toxicity in patients with peripheral blood malignant B-cell lymphocytosis. AB - Infusion related adverse events (AE) with day 1 rituximab in patients with B-cell non-Hodgkin's lymphoma (NHL) are common. The purpose of this study was to evaluate the AE occurring in patients with malignant B-cell lymphocytosis who received rituximab. Patients with a > or = 3 x 10(9)/L absolute lymphocyte count (ALC) receiving rituximab from 1998 to 1999 or participating in a phase I study of rituximab and interleukin-12 were reviewed. The AE occurring on the day of rituximab, the treatment provided (including hospitalization), and the subsequent ALC responses were recorded. Twenty-seven patients were identified; 14 had NHL, one Waldenstrom's macroglobulinemia, and 12 patients had chronic lymphocytic leukemia. The baseline median ALC was 9.58 x 10(9)/L (mean, 49.31; range, 3.56 380.95). All patients received rituximab as an outpatient. There were only two AE > or = grade 3. One patient was hospitalized for 1 day for i.v. fluids to treat an increase in creatinine that occurred with tumor lysis. A second patient developed a pulmonary syndrome five days after day 1 rituximab and required mechanical ventilation, but had no long-term lung toxicity. This study demonstrates that patients with high numbers of circulating blood B-lymphocytes can usually safely receive rituximab as outpatients. Patients who experience a rapid drop in ALC should be monitored closely for tumor lysis and the pulmonary syndrome. PMID- 11911417 TI - Cyclosporin A for the treatment of pure red cell aplasia associated with myelodysplasia. AB - Recent reports have highlighted an associated of pure red cell aplasia (PRCA) with myelodysplasia (MDS). There are minimal data on the response of PRCA in this context to immunosuppression; in particular the role of cyclosporin A (CSA) has not been evaluated. We describe a patient with PRCA/MDS in whom CSA and low dose prednisolone led to restoration of marrow erythropoietic activity. Tests indicated the PRCA was likely to be mediated by cytotoxic lymphocytes rather than an inhibitory effect of cytokines. These observations suggest that CSA, due to its suppressive effect on cytotoxic T cell activity, is a rational therapy in this context. PMID- 11911418 TI - A polymorphism in the BCL-6 gene is associated with follicle center lymphoma. AB - Follicle center lymphoma (FCL) accounts for approximately 40% of all non Hodgkin's lymphomas (NHL). The genetic-environmental interactions involved in the etiology and pathogenesis of this disease are unknown. In our previous study a single nucleotide polymorphism (SNP) (397C) in the regulatory untranslated first intron region of the BCL-6 gene was found in four of the eight FCL patients but in none of the 10 healthy controls. To further evaluate the potential association between the 397C allele of the BCL-6 gene and FCL, we performed a case-control study. Genomic DNA was isolated from 85 FCL patients, from 98 control cases without a previous history of malignancy, treated at Stanford University Medical Center for non-malignant disorders and from 90 samples from the DNA Polymorphism Discovery Resource. The 397G and the 397C polymorphic alleles were identified by a PCR-RFLP method. To evaluate the possible effect of this polymorphism on gene expression, BCL-6 mRNA levels in nine FCL tumors with the 397G-G genotype and in nine FCL tumors with the 397G-C genotype were measured by quantitative real-time RT-PCR. The 397C polymorphic allele was found in 32 FCL cases (37.6%), in 20 controls (20.4%) and in 17 (18.9%) samples from the DNA Polymorphism Discovery Resource. The prevalence of the 397G-C and 397C-C genotypes was significantly higher in FCL cases than in control group (p = 0.01). No difference in BCL-6 gene expression was observed between FCL cases with 397G-G and 397G-C genotypes. The present study demonstrates a possible association between the 397C allele of the BCL-6 proto-oncogene and FCL. The similar levels of BCL-6 mRNA expression in 397G G and in 397G-C FCL cases suggests that any possible oncogenic effect of the polymorphic allele would not simply be related to a direct effect on BCL-6 gene expression and suggests the existence of other FCL susceptibility genes that are in linkage disequilibrium with the 397C allele of the BCL-6 gene. PMID- 11911419 TI - Expression of G-CSF receptor on myeloid progenitors. AB - Although granulocyte colony-stimulating factor (G-CSF) has been reported to act on cells of neutrophilic lineage, the expression of receptors for G-CSF (G-CSFR) on human hematopoietic progenitor cells has been unclear. We then analyzed the expression of G-CSFR on human bone marrow and G-CSF mobilized peripheral blood CD34+ cells, and examined the proliferation and differentiation capabilities of sorted CD34+ G-CSFR+ and CD34+ G-CSFR- cells using methylcellulose clonal culture. These results indicate that the expression of G-CSFR on CD34+ cells is restricted to myeloid progenitors, suggesting that the specific activity of G-CSF on myelopoiesis depends on the exclusive expression of its receptor on myeloid progenitors, and that the mobilization of various hematopoietic stem/progenitor cells is not a direct effect of G-CSF in humans. PMID- 11911420 TI - The presence of circulating clonal CD19+ cells in multiple myeloma. AB - Multiple myeloma (MM) is a B-cell malignancy characterized by the expansion of mature plasma cells (PC) localized in the bone marrow (BM). Several studies have identified circulating clonotypic CD19+ cells at a differentiation stage preceding the PC. The level of circulating clonotypic CD19+ cells is highly variable but generally low. Circulating clonotypic cells respond well to induction therapy, although a small subset within the CD19 compartment is resistant even to high-dose chemotherapy. The clonal CD19+ cells represent an ongoing differentiating population ranging from memory B-cells to plasmablasts. However, a clonal relationship gives no proof of malignant potential, and whether or not clonotypic precursor cells are involved in the disease process is a subject of intense debate. Translocations involving the immunoglobulin locus (14q32) are an early non-transforming event common to both monoclonal gammopathy of undetermined significance (MGUS) and MM introduced at the memory B-cell level. At the plasmablast stage, a phenotypic transformation occurs with downregulation of CD19 and upregulation of myeloma specific markers such as CD56, CD117 and CD28. Translocations involving the isotype-switch machinery and the introduction of tumor-specific markers at the plasmablast stage suggest that the clonal CD19+ memory B-cells and CD19+ plasmablasts are non-malignant, but immortalized relatives that gave rise to myeloma. A final proof of the malignant potential of CD19+ clonotypic cells might await the identification of the molecular events causing the transformation in myeloma. PMID- 11911421 TI - CD40 ligand immunotherapy in cancer: an efficient approach. AB - Cancer cells do not elicit a clinically sufficient anti-tumor immune response that results in tumor rejection. Recently, many investigators have been trying to enhance anti-tumor immunity and encouraging results have been reported. This review will discuss current anti-cancer immunotherapy; interleukin-2 therapy, tumor vaccine secreting Granulocyte macrophage-colony stimulating factor, dendritic cells fused with tumor cells, and CD40 ligand immunotherapy. Moreover, we introduce our two kinds of CD40 ligand immuno-genetherapy; (1) oral CD40 ligand gene therapy against lymphoma using attenuated Salmonella typhimurium (published in BLOOD 2000), (2) cancer vaccine transfected with CD40 ligand ex vivo for neuroblastoma (unpublished). Both approaches resulted in a high degree of protection against the tumor progression and they are simple and safe in the murine system. PMID- 11911422 TI - Immunophenotype changes and loss of CD52 expression in two patients with relapsed T-cell prolymphocytic leukaemia. AB - T-cell prolymphocytic leukaemia (T-PLL) is an aggressive disease often resistant to conventional chemotherapy. Long lasting remissions with the monoclonal antibody CAMPATH-1H (anti-CD52) have been documented. We describe two unusual T PLL patients treated successfully first with CAMPATH-1H in whom, at the time of relapse, the cells underwent a phenotypic switch with loss of CD52 expression. In one of them, cytogenetic analysis demonstrated the same chromosome abnormalities in the cells at diagnosis and relapse. The reasons for the immunophenotypic changes are unknown but it is likely that loss of CD52 antigen expression contributed to the resistance to CAMPATH-1H in one of the patients when re treated. PMID- 11911423 TI - Aggressive natural-killer cell lymphoma presenting with skin lesions, breast nodule, suprarenal masses and life-threatening pericardial and pleural effusions. AB - We report the clinical and laboratory findings of a patient with an aggressive Epstein-Barr virus positive CD2+/CD56+ natural killer-cell lymphoma with a high mitotic activity and complex chromosomal abnormalities presenting with life threatening pericardial and pleural effusions, disseminated skin lesions, breast nodule and large suprarenal masses. The clinical course was characterized by resistance to chemotherapy and relapsing pericardial and pleural effusions with respiratory and haemodynamic failure. Death occurred 4 months after the first manifestations of the disease as a consequence of cardiac tamponade. PMID- 11911424 TI - Cure of pulmonary Rhizomucor pusillus infection in a patient with hairy-cell leukemia: role of liposomal amphotericin B and GM-CSF. AB - We describe a case of successfully treated multifocal pulmonary Rhizomucor pusillus, a condition which has previously been universally fatal. A 77 year-old man had a background of chronic neutropenia due to hairy-cell leukemia, splenectomy, corticosteroid therapy and an obstructing left ureteric transitional cell carcinoma. He was successfully treated with 3 months of high-dose liposomal amphotericin B and 7 months of granulocyte-macrophage colony-stimulating factor. Treatment was complicated by mild reversible deterioration of renal function. There was a near complete radiological response to the therapy at 6 months and the patient remains well 20 months following diagnosis of R. pusillus and 13 months following cessation of treatment. PMID- 11911425 TI - Hemophagocytic syndrome associated with inappropiate secretion of antidiuretic hormone in lymphoma and acute myeloblastic leukemia: report of two cases. AB - Hemophagocytic syndrome (HPS) is a rare clinicopathological disorder characterized by systemic proliferation of phagocytizing histiocytes associated with fever, cytopenias, lymphadenopathy, hepatosplenomegaly, and disseminated intravascular coagulopathy. We present the association of hemophagocytic syndrome associated with inappropriate secretion of antidiuretic hormone (SIADH) in two cases of hematological malignancies; anaplastic large cell lymphoma (ALCL) and acute myeloblastic leukemia (AML M4) In the patient with lymphoma, the diagnosis of lymphoma, HPS and SIADH were concurrent. In the patient with AML, HPS and SIADH were observed while the patient was in hematological remission. Thus it seems that patients with HPS may also carry a risk for the development of SIADH; the relationship with HPS and SIADH should be further investigated. PMID- 11911426 TI - Monotherapy with rituximab induces rapid remission of recurrent cold agglutinin mediated hemolytic anemia in a patient with indolent lympho-plasmacytic lymphoma. AB - Cold agglutinin mediated immune hemolytic anemia secondary to lymphoproliferative disease (LPD), is primarily treated with measures directed to eliminate the malignant clone and as such, chemotherapy is usually given. The recent availability of monoclonal antibodies, has made it feasible to obtain both a clinical and molecular remission, as well as a remission on the functional level, such as elimination of secondary autoimmune phenomena. Recently we have administered a course of monotherapy with rituximab (4 weekly injections, x 375 mg/m2) to a patient with refractory and transfusion dependent cold agglutinin mediated hemolytic anemia secondary to indolent B-cell lymphoma. She achieved complete remission with a significant improvement in hemolysis and also became transfusion independent with a current follow-up of over one year. In individual cases, Rituximab has the potential of achieving not only a complete clinical remission (CR) but also a molecular CR, as well as a "functional" CR, by eliminating the clinical manifestations of autoimmunity; in this case, cold agglutinin mediated hemolytic anemia, secondary to NHL. Good results in autoimmunity secondary to lymphoma raises the possibility of future potential benefit of this agent in other primary autoimmune disorders. PMID- 11911427 TI - Isolated pentasomy of chromosome 8 in erythroleukemia. AB - Pentasomy 8 as a sole anomaly in hematological disorders is rare. Only 2 such cases, one in acute monocytic leukemia and one in chronic myelomonocytic leukemia have been described in the literature to date. Here, we report the first case of a 42 year old man with erythroleukemia displaying a pentasomy 8 clone. Conventional cytogenetics of bone marrow cells showed 16 metaphases with pentasomy 8 and 9 with normal diploidy. Fluorescence in situ hybridization (FISH) analysis using a whole chromosome painting probe and a centromeric probe specific for chromosome 8 confirmed the presence of pentasomy 8 and also revealed a low percentage of a trisomic and a tetrasomic clone. The patient died three days after diagnosis without chemotherapy. The findings suggest that pentasomy 8 is associated with a heterogeneous group of myeloid disorders and probably plays a specific role in the progression of myeloid neoplasia. PMID- 11911428 TI - Clonality of acquired primary pure red cell aplasia: effectiveness of antithymocyte globulin. AB - Primary pure red cell aplasia (PRCA) was diagnosed in two male patients, 65 and 69 years old respectively. In both, surface markers of peripheral blood nuclear cells revealed the presence of TCR alphabeta+ phenotype. Clonality of T cells was confirmed by the polymerase chain reaction in both patients, in whom, prednisone at a dose of 1 mg/kg/day improved the anemia and lower doses caused its renewal, resulting in the reappearance of the patient's transfusion requirement. On the other hand, the anemia seems to have been treated permanently (second case) with horse antithymocyte globulin (ATG) (20 mg/kg/day 1 to 8 +) since his hemoglobin was about 15 g/dl at the time of writing. In the first patient, the hemoglobin level was 10.5 g/dl one month after the administration of ATG (15 mg/kg/d 1 to 5 +), but unfortunately, the patient died because of a massive gastrointestinal bleeding on the fortieth day following this treatment. We, therefore, suggest that, patients with acquired primary PRCA should be screened to detect the presence of a T-cell clone and recommend that, treatment should start earlier with ATG, if the PRCA is due to a T-cell clonal disorder. PMID- 11911429 TI - Primary anaplastic large cell lymphoma of the spleen presenting as a splenic abscess. AB - Large cell anaplastic lymphoma (ALCL) is characterized mainly by the presence of large, atypical lymphoid cells with anaplastic nuclear morphology and positivity to Ki-1 antigen. We describe, to our knowledge, the fourth reported case of primary ALCL of the spleen. The patient, a 62 year old woman, presented symptoms resembling a splenic abscess. PMID- 11911430 TI - Jejunal perforation in a patient with adult T-cell leukemia. AB - We present a case of adult T-cell leukemia (ATL) with jejunal perforation at the site of intestinal involvement by ATL. A 39-year-old woman presented with sudden onset abdominal pain. Physical examination showed generalized severe abdominal tenderness and intraabdominal free air was seen on radiographic examination. Under a diagnosis of peritonitis due to intestinal perforation, an emergency operation was performed. A pinhole-like perforation was found in the jejunum 80 cm distal to Treitz's ligament, and the patient underwent partial resection of the affected jejunum. Microscopic examination revealed diffuse infiltration of abnormal lymphocytes into the entire wall of the jejunum and mesenteric lymph nodes. A diagnosis of ATL was confirmed by the presence of antibody to human T lymphotropic virus type 1 (HTLV-1) in the serum, a positive result for T-cell markers and the HTLV-1 proviral genome in the mononuclear cells in the specimens. The final diagnosis was thus lymphoma subtype of ATL. Combination chemotherapy was repeated until the patient died 14 months postoperatively. Emergent surgery followed by intense chemotherapy might improve survival in patients with ATL and perforated intestine. PMID- 11911431 TI - Focal pulmonary uptake of gallium-67 due to radiation pneumonitis: the case for a misdiagnosis of Hodgkin's disease progression. AB - Gallium-67 scan is usually performed in patients with Hodgkin's disease and high grade non-Hodgkin lymphoma for evaluation of disease status after treatment. We present a case of an asymptomatic woman in complete remission of Hodgkin's disease after treatment with chemotherapy and radiotherapy where a focal uptake of Gallium-67 was discovered two months after finishing treatment. As classical radiation pneumonitis can appear one to three months after finishing radiotherapy and normally has an asymptomatic course, this possibility should be considered in these cases, especially when prior chemotherapy was administered. PMID- 11911432 TI - Early invasive pulmonary aspergillosis in a leukemia patient linked to aspergillus contaminated marijuana smoking. AB - 46-year-old patient with acute myeloid leukemia (AML) whose disease manifested as fever, chills and dry cough is reported here. Despite broad antibiotic coverage he remained acutely ill with spiking fever, shaking chills, and hypoxemia. His initial chest radiograph was normal but chest computed tomography (CT) scan disclosed bilateral focal infiltrates. Hypoxemia and severe thrombocytopenia precluded invasive diagnostic procedures. A thorough epidemiological investigation revealed that before becoming acutely ill the patient smoked daily tobacco mixed with marijuana from a "hookah bottle". While waiting for tobacco and "hookah water" cultures, we started antifungal therapy. Resolution of fever and hypoxemia ensued after 72 hours. Tobacco cultures yielded heavy growth of Aspergillus species. We suggest that habitual smoking of Aspergillus-infested tobacco and marijuana caused airway colonization with Aspergillus. Leukemia rendered the patient immunocompromised, and allowed Aspergillus to infest the lung parenchyma with early occurrence of invasive pulmonary aspergillosis. Physicians should be aware of this potentially lethal complication of "hookah" and marijuana smoking in immunocompromised hosts. PMID- 11911433 TI - Pure red cell aplasia associated with parvovirus B19 infection in T-large granular lymphocyte leukemia. AB - There have been few reports of large granular lymphocyte (LGL) leukemia with neutropenia complicated with pure red cell aplasia (PRCA) that developed after human parvovirus (HPV) B19 infection. We report here the case of a 35-year-old female who developed HPV B19-associated PRCA with T-LGL leukemia. LGL count of peripheral blood was lower than 2 x 10(9) l(-1), although phenotypic analysis of LGL showed CD3+, CD16-, CD56-, CD57+ with double positive for CD3 and CD57, and genetic study showed the clonal rearrangement of T-cell receptor gene. Microscopically, the patient's bone marrow showed characteristic giant proerythroblasts. A serologic study of HPV B19 was positive for IgM, but negative for IgG, with a positive result on Dot-blot hybridization assay for HPV B19 DNA. Severe anemia and reticulocytopenia ameliorated without treatment 10 days after the initial examination, but slight anemia, neutropenia, a moderate increase of LGL counts with rearrangement of TCR gene, and positive result of HPV B19 DNA has persisted 7 months after the initial examination. We suggest that this viral infection may play an etiologic role in some patients with LGL leukemia who develop PRCA. PMID- 11911435 TI - Terminology for sample collection in clinical genetic studies. PMID- 11911434 TI - G-CSF related capillary leak syndrome in a child with leukemia. PMID- 11911436 TI - Pharmacogenomics and the (ir)relevance of race. PMID- 11911437 TI - Pharmacogenomics and schizophrenia: clinical implications. PMID- 11911438 TI - Global perspectives on proteins: comparing genomes in terms of folds, pathways and beyond. AB - The sequencing of complete genomes provides us with a global view of all the proteins in an organism. Proteomic analysis can be done on a purely sequence based level, with a focus on finding homologues and grouping them into families and clusters of orthologs. However, incorporating protein structure into this analysis provides valuable simplification; it allows one to collect together very distantly related sequences, thus condensing the proteome into a minimal number of 'parts.' We describe issues related to surveying proteomes in terms of structural parts, including methods for fold assignment and formats for comparisons (eg top-10 lists and whole-genome trees), and show how biases in the databases and in sampling can affect these surveys. We illustrate our main points through a case study on the unique protein properties evident in many thermophile genomes (eg more salt bridges). Finally, we discuss metabolic pathways as an even greater simplification of genomes. In comparison to folds these allow the organization of many more genes into coherent systems, yet can nevertheless be understood in many of the same terms. PMID- 11911439 TI - Critically assessing the state-of-the-art in protein structure prediction. AB - One of the most tantalising 'grand challenges' in structural biology is to solve the problem of predicting the structure of a protein from its amino acid sequence alone. Although this problem appeals to many researchers on a purely academic level, the practical importance of protein structure prediction has become particularly clear with the release of the first draft of the complete human genome sequence last year. This moved modern biology into the new so-called 'post genome' era, and for the foreseeable future, one of the main issues in modern biology will be the characterisation of the many 'unknown' gene sequences which are now sitting waiting in DNA and protein data banks. Protein structure can provide a great deal of insight into the evolutionary origins, function and mechanism of a protein, and so any means for determining the 3-D structure of a novel protein will likely be of critical importance. PMID- 11911440 TI - Genes that co-cluster with estrogen receptor alpha in microarray analysis of breast biopsies. AB - The estrogen receptor plays a critical role in the pathogenesis and clinical behavior of breast cancer. To better understand the molecular basis of estrogen dependent forms of this disease we studied gene expression profiles from 53 primary breast cancer biopsies. Gene expression data for more than 7000 genes were generated from each tumor sample with oligo microarrays. A standard correlation-clustering algorithm identified 18 genes that co-clustered with estrogen receptor alpha. Eleven of these genes had previously been associated with estrogen regulation or breast tumorigenesis including trefoil factor 1 and estrogen regulated LIV-1. Additional study of these 18 genes may further delineate the role of estrogen receptor in breast cancer, generate new predictive biomarkers for response to endocrine therapies and identify novel therapeutic targets. PMID- 11911441 TI - Accuracy of two-dimensional electrophoresis for target discovery in human colorectal cancer. AB - Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) is increasingly used for target discovery in human disease to complement genomic studies. We have assessed the possibilities and limits of 2-D PAGE applied to human colorectal cancer. Up to 10(8) epithelial cells were purified from paired normal and pathological biopsies using Ber-EP4 coated magnetic beads, allowing the elimination of cellular and fluid contaminations. The mean coefficient of variation (CVAR) of repeated 2-D PAGE analysis with silver staining was lying between 20 and 28%. However, only 47% (interrun) to 76% (intrarun) of spots could be matched within a triplicate experiment. Interindividual phenotypic variability was high. Intratumoral phenotypic variability was not found to be significant. When method and tumor variability were added, 90% of CVAR were inferior to 48%. Thus, two-fold up- or down-regulation of protein expression reveals biological significance. Serial paired comparison of 923 proteins in 10 patients showed highly reproducible differences between normal and cancer tissues. Under well defined experimental conditions and after the high variability of the technique has been considered, 2-D PAGE parallel analysis of paired colorectal samples allows patient-specific tumor profiling. PMID- 11911442 TI - The VNTR polymorphism of the human dopamine transporter (DAT1) gene affects gene expression. AB - The human dopamine transporter (DAT1) gene contains a variable number of tandem repeats (VNTR) in its 3'-untranslated region (UTR). The linkage and association between the VNTR polymorphism of DAT1 and various neuropsychiatric disorders have been reported. We have determined the genomic structure of DAT1 genes containing 7-, 9-, 10-, and 11-repeat alleles and examined the effect of VNTR polymorphism in the 3'-UTR region of DAT1 on gene expression using the luciferase reporter system in COS-7 cells. Luciferase expression was significantly higher when the 3' UTR of the DAT1 gene contained the 10-repeat allele than when it contained the 7- or 9-repeat alleles. This suggests that VNTR polymorphism affects the expression of the dopamine transporter. PMID- 11911443 TI - Pharmacogenomics and ethnic minorities. PMID- 11911445 TI - RAGE and PRAY. PMID- 11911444 TI - Mining the bibliome. PMID- 11911446 TI - Is annexin 7 a tumor suppressor gene in prostate cancer? PMID- 11911447 TI - A critical assessment of the role of pharmacogenomics and pharmacogenetics approaches to cardiovascular diseases. PMID- 11911448 TI - Time flies like an arrow. Fruit flies like crack? PMID- 11911449 TI - Length of marriage and its effect on spousal concordance in Kuwait. AB - It was hypothesized that marriage duration affects physical and cultural homogamy and spousal concordance in Kuwaiti marriages. Westernization increased spousal correlations due to fewer arranged marriages and increased individual spousal choice. Spousal similarities for selected physical and cultural traits were also examined for couples married 15 years or less, 16 to 30 years, and 31 years and more. Consanguineous couples belong to the al-Kandari, one of the largest and most important kindreds in Kuwait, who traditionally married kin and continue to do so. Six physical measurements and blood pressure were taken along with a sociocultural questionnaire to examine cultural preferences. In all, 242 couples (484 people) participated; 62 couples were in non-consanguineous unions. It was hypothesized that in shorter-duration unions spouses would be more alike for physical and cultural traits. For physical traits, results for stature, weight, the body mass index, and hip circumference are congruent with the hypothesis, whereas results for the triceps and subscapular skinfolds, waist circumference, and systolic and diastolic blood pressure are not. Overall, for some traits spouses are more alike than in previous generations, and specific aspects of similarity among long-term spouses reflect historical and cultural phenomena. PMID- 11911450 TI - Fat distribution in Venezuelan children and adolescents estimated by the conicity index and waist/hip ratio. AB - This study compares the conicity index (C) with the waist/hip ratio (WHR) in a cross-sectional sample of Venezuelan children (n = 784 boys and n = 735 girls), 3 to 16 years of age. Distributions of C and WHR were compared in Box-plot diagrams. Regression analysis was used to examine the relationship between indices by age and sex. Conicity captured more outliers in the distribution than WHR and explained 33% to 62% of the variability in WHR in three age groups. The influence was stronger in females during adolescence (R2 = 0.60, P < 0.05). According to the principle of C. most children presented a bi-conical shape, which was more pronounced in boys than girls and which was indicative of a more central distribution of adiposity. These results are related, in part, to age and sex differences in body composition and to the earlier onset of the adolescent growth spurt in Venezuelan children. PMID- 11911453 TI - Stature and stature distribution in Portuguese male adults 1904-1998: the role of environmental factors. AB - The records of height of 841,457 18-year-old Portuguese males were analyzed by area of residence (districts). The sample included all Portuguese 18-year-old males born between 1966 and 1979 and examined between 1985 and 1998, in the north, center, and south of Portugal. They represent all social strata. There were statistically significant differences (P < or = 0.001) among districts: males from Lisbon (172.8 cm) and Setubal (172.7 cm), the most developed districts, were tallest, and those from Madeira (169.7 cm) and Coimbra (171.6 cm) were shortest. Compared with published data for 1904, there was a positive secular trend in height. The average increase was 8.93 cm and the estimated rate was 0.99 cm per decade. The changes that occurred were mainly the result of the reduction of the shortest classes of stature, those < 150 cm to 170 cm, and an increase in the frequency of the highest classes, > or = 170 cm. This positive trend and the changes in stature distribution must be related to the general improvement in standard living conditions that occurred in Portugal primarily after the 1960s and 1970s, especially in terms of nutrition and the health system. Taking into account the socioeconomic differences that still exist between districts, the results suggest that the secular trend in height should continue for the Portuguese population in future decades. PMID- 11911451 TI - Genetic variability in the Guahibo population from Venezuela. AB - Four communities from Guahibo of Venezuela were analyzed for the genetic variants of nine erythrocyte enzymes and five serum proteins. Of the 14 loci determined, four were monomorphic. Significant frequency differentiation among communities, was present for ESD and TF markers. In general, Guahibo allele frequencies are in the variation ranges described for South American groups. The analysis indicates a relatively higher affinity of Guahibos with other Venezuelan groups within an irregular pattern of genetic distances that are likely related to the complex demographic history of the South American groups. Genetic diversity estimates reveal a moderate degree of genetic structure between the four Guahibo communities. This intra-tribal variability in Guahibo appears to be lower than in Venezuelan Piaroa but higher than in other Amerindians and could be attributed to a combined effect of low population size and relative isolation of communities. At a continental level, the distribution of genetic diversity is consistent with preferential population movements along the eastern and western coastal areas. PMID- 11911452 TI - Anthropometric characteristics of pregnant women in Cali, Colombia and relationship to birth weight. AB - Anthropometric dimensions were collected from 46 pregnant women living in Cali, Colombia to gain a better understanding of how poor, urban women deal with the demands of pregnancy and to identify relationships between maternal characteristics and infant birth weight. Height, weight, skinfold thicknesses (subscapular, suprailiac, thigh, calf, and triceps), and circumferences (hip, thigh, calf, and mid-upper arm) were measured on all women. Infant measurements were weight and length. The women were measured in the second and third trimesters, and a subsample (n = 16) was measured twice in the third trimester. Mean birth weight was 3,137.6 +/- 488.5 g (n = 44), and mean length was 49.8 +/- 3.0 cm. All but three of the infants were full-term, and the incidence of low birth weight (LBW) was 9%. The 46 women showed a significant increase in weight (P < 0.001); subscapular, suprailiac, and mid-thigh skinfold thicknesses (P < or = 0.01) and in hip, thigh, and calf circumferences (P < or = 0.01) between trimesters 2 and 3. Women who gave birth to both normal birth weight (NBW) and LBW infants showed significant increases in weight (P < 0.001 and P = 0.02, respectively), but only women who had NBW infants showed significant increases in the suprailiac skinfold and hip circumference (P < 0.001). In the third trimester, attained weight, skinfold thicknesses, and circumferences tended to be greater in women who had NBW infants. In general, this group of women gained less weight and had a greater incidence of LBW infants compared with women in developed countries, but changes in skinfold thicknesses over the course of pregnancy were similar compared with other studies. PMID- 11911454 TI - Human growth in southern Zambia: a first study of Tonga children predating the Kariba Dam (1957-1958). AB - During the late 1950s the Kariba hydro-electric dam was constructed on the border of Zambia and Zimbabwe forcing the relocation of 57,000 people, mainly Tonga. As part of a larger study to assess the effects of the relocation, research into the human biology of the Tonga people was conducted. The research reported here provides a basis for comparison with long-term follow-up data on growth and physical status of Gwembe Tonga to determine the effects of resettlement. The sample consists of 303 schoolchildren, 7 to 13 years, from two schools that were not relocated and three schools that were to be relocated. Homogeneity of the two groups led us to combine them as a single baseline sample before relocation. Comparisons with NCHS (National Center for Health Statistics) reference data and with contemporary urban data from southern Zambia indicate sub-optimal nutritional status. After ages 6 and 7, height-for-age and weight-for-age Z scores of boys decline steadily towards -2.0 SD throughout the 12th year, whereas mean Z-scores of girls decline markedly from 8 years on. Mean HAZ (Height-for-age Z-score) of girls falls below -2.0 SD by 11 years and approaches -3.0 SD by 13 years. Thirty-nine percent of males and 47% of females in the baseline sample showed effects of moderate or severe protein energy malnutrition at the time of relocation. Because these children were from a school sample, gender differences in opportunity to attend school may be a factor in what appears to be gender bias, favoring the status of boys. This sample provides a baseline for assessing the long-term impact of forced relocation on the Gwembe Tonga. PMID- 11911455 TI - Evidence of age-related responses to short-term environmental variation: time series analysis of cross-sectional data from Taiwan, 1969 to 1990. AB - Cross-sectional heights and weights collected by Taiwan's Ministry of Education since 1964, and reported separately for Taipei and rural townships from 1969 through 1990, were used to test the hypothesis that in Taiwan the degree to which increments of change in height and weight of urban or rural boys and girls tracked each other from year to year was a function of sex and age-related capacities to respond to common environmental forces. Five testable implications follow from this hypothesis. Results were evaluated using detrended time series derived correlations of yearly or near yearly change in sex-specific age group height or weight means within and across regions, cross-sectionally estimated ages of maximum change of height and weight, and evidence of response to two recessions. The hypothesis was largely supported by the available evidence. The similarities in secular change from year to year, with only a few exceptions, suggested that socioeconomic trends indirectly affected the pace of growth of children in both urban and rural Taiwan measured in the spring of the following year. The strength and age-related pattern of these correlations were consistent with sex and age-related capacities to respond to shared environmental factors. Evidence of lags between height and weight, found both in cross-sectionally estimated mean ages of maximum change in height and weight and in the pattern of within sex correlations also argued for the hypothesis. The evidence further suggested that when the economy faltered briefly in Taiwan following worldwide oil crises, children's growth was rapidly affected in an age-dependent manner. This study bolsters the view that secular changes in growth reflect environmental quality. It suggests as well that under some circumstances, environmental change may be rapidly reflected as mean changes in height or weight of certain age groups. The intensity of the response appears to be related to the degree of improvement or deprivation, the maturationally mediated pace of growth, and probably initial energy balance. PMID- 11911456 TI - Prediction of cross-sectional geometry from metacarpal radiogrammetry: a validation study. AB - Regression models have been developed to adjust algebraic estimates of second metacarpal cortical bone geometry to actual values (as determined through invasive analysis). These models, derived from an archaeological sample of European origin, have high efficacy in predicting actual values but have not been validated on non-European samples. This paper reports a validation study for these models applied to a historic/proto-historic sample of Inuit from the central Canadian Arctic (n = 166; ages and sexes pooled as per the original study). In that the Inuit sample has been argued to exhibit distinct skeletal biology, this represents a robust test of the predictive models. The algebraic models again produced biased overestimates of actual values, whereas the predictive regression models were found to provide good estimates of actual values for measures of bone strength (Total Area, bending about the Ix and Iy axes), but not for estimates of mass (Cortical Area). This difference may exist in either functional or systemic differences in skeletal physiology and aging bone loss in the Inuit. PMID- 11911457 TI - Variation at 10 protein coding loci in the Mbenzele Pygmies from the Central African Republic and a comparison with microsatellite data. AB - Ten protein coding loci (6-PGD, A1-AT, ACP1, CaII, ESD, GC, GPX1, Hb beta, PGM1, and TF) were analyzed in the Mbenzele Pygmies from the Central African Republic. The frequency data were used to calculate the genetic distances between Mbenzele Pygmies and other African groups. In the principal coordinate plot of FST genetic distances, the Mbenzele cluster together with other Pygmies of the western cluster, the Biaka from C.A.R., Gielli from Cameroon, and Babinga from Congo. By contrast, they are considerably distanced from other Pygmy groups of the eastern cluster (Twa from Rwanda, Mbuti from Zaire). Genetic distances obtained using protein loci were compared with those based on microsatellite loci. The two distance matrices are insignificantly correlated (r = 0.268; one tail probability = 0.332), and the main difference is in the higher genetic affinity between the Mbenzele and Biaka Pygmies observed at the protein level. Although reasons underlying the discrepancy between inter-populational variation at protein and DNA loci are not established with certainty, the comparison suggests that the genetic distance between the Mbenzele and Biaka Pygmies at microsatellite loci could have been shaped by genetic drift. PMID- 11911458 TI - Functional genomics of the social amoebae, Dictyostelium discoideum. AB - Dictyostelium discoideum is one of the simplest organisms to form a multicellular structure, and it offers several advantages as a model. In order to understand the genetic basis of the multicellular development, a comprehensive analysis of cDNAs is being performed. To date, about 75,000 ESTs have been collected at different stages of development. They have been assembled into about 6,400 independent sequences that represent 70-80% of all of the expected genes in D. discoideum. The results are available on the Internet. In addition to structural analyses, functional analyses of the temporal and spatial expression patterns and gene targeting are being carried out. Furthermore, there are plans to combine the information that is obtained from the cDNA, Genome, and Proteome Projects, as well as the published results, into an integrated database, DictyBase. PMID- 11911460 TI - Inhibitory effect of zeatin, isolated from Fiatoua villosa, on acetylcholinesterase activity from PC12 cells. AB - Acetylcholinesterase (AChE) inhibitors, which enhance cholinergic transmission by reducing the enzymatic degradation of acetylcholine, are the only source of the compound that is currently approved for the treatment of Alzheimer's disease (AD). The methanol extract from Fiatoua villosa among 100 traditional edible plants that were tested, showed the most potent inhibitory effect (51%) on acetylcholinesterase in vitro. After the sequential solvent fractionation of the methanol extract of Fiatoua villosa, the active fraction was repeatedly subjected to open-column chromatography on silica gel. From the highest inhibitory fraction, the chloroform fraction (75%) on AChE, the single compound, was obtained by the Sep-Pak Cartridge (C18: reverse phase column). This compound was finally purified by HPLC (micro-bondapack C18 reverse phase column: 19 x 300 mm). According to the electron impact mass spectrometry (EI-MS), we confirmed that the molecular mass was 219 m/z. The structure of this compound was identified as zeatin [2-methyl-4-(1H-purine-6-ylamino)-2-buten-1-ol], one of the derivatives of purine adenine. The concentration that was required for 50% enzyme inhibition (IC50 value) was 1.09 x 10(-4) M. This study demonstrated that the zeatin from Fiatoua villosa appeared to be the most potent AChE inhibitor in AD. PMID- 11911459 TI - Protection by carnosine-related dipeptides against hydrogen peroxide-mediated ceruloplasmin modification. AB - Carnosine, homocarnosine, and anserine are present in high concentrations in the muscle and brain of many animals and humans. Previous studies showed that these compounds have an antioxidant function. We investigated the protective effects of carnosine and related compounds on the modification of human ceruloplasmin that is induced by H2O2. Carnosine, homocarnosine, and anserine significantly inhibited the fragmentation and inactivation of ceruloplasmin that is induced by H2O2. All three compounds also inhibited the release of copper ion from protein, and the formation of hydroxyl radicals in the ceruloplasmin/H2O2 system. These compounds inhibited the fragmentation of human serum albumin that is induced by the copper-catalyzed oxidation system, as well as by the iron-catalyzed oxidation system. These results suggest that carnosine, homocarnosine, and anserine might protect ceruloplasmin against H2O2-mediated oxidative damage through a combination of copper chelation and free radical scavenging. PMID- 11911461 TI - Refolding of the catalytic and hinge domains of human MT1-mMP expressed in Escherichia coli and its characterization. AB - The catalytic and hinge domain (Tyr112-Ile318) of the human membrane type-1 matrix metalloproteinase (MT1-MMP; MMP-14), containing hexa-histidines at the C terminus (chMT1-MMP), was overexpressed in Escherichia coli. The expressed polypeptide was almost exclusively found in the inclusion body, and then purified by a single Ni2+-NTA agarose column chromatography after solubilization with 6 M urea. During refolding, the 26.9 kDa chMT1-MMP was processed to a 24.3 kDa intermediate form and then to a 22.2 kDa mature form. By Western blot analysis and mass spectrometry combined with N-terminal sequencing, the intermediate form was identified as a mixture of the Tyr112-Thr299 with a translation-initiating methionine and Ile114-Thr299, and that the mature form corresponds to Ile114 Pro290. These results demonstrate that the mature form was generated by successive autoproteolysis of the N- and C-terminal sites between Thr299-Thr300, Ala113-Ile114, and Pro290-Thr291 during refolding. Catalytic activity of the mature chMT1-MMP was demonstrated by a peptide cleavage assay. In addition, it has gelatinolytic activity and is able to activate proMMP-2 to the mature MMP-2. These results indicate that the refolded chMT1-MMP retains characteristics of MT1 MMP. PMID- 11911462 TI - A cDNA clone for cyclophilin from Griffithsia japonica and phylogenetic analysis of cyclophilins. AB - A cDNA clone, designated as Griffithsia japonica cyclophilin-1 (GjCyp-1), was isolated by differential screening of a cDNA library for a red alga, G. japonica. The transcript that corresponded to GjCyp-1 was abundant in vegetative, male, and tetrasporangial thalli, but only the basal level of the transcript was detected in female gametophytes. Determination of the nucleotide sequence of GjCyp-1 identified an open reading frame (ORF), which shared high homologies with cyclophilins that were previously reported in other organisms. Currently available amino acid sequences of eukaryotic cyclophilins were compared in order to examine their phylogenetic relationship to GjCyp-1. A phylogenetic analysis, based on the aligned sequences, showed two major clades - cytosolic cyclophilins (CypA) and ER cyclophilins (CypB). The clade of CypA was divided into six groups plant, nematode, mammal, euglenozoa, fungi, and platyhelminthes CypA. GjCyp-1 appeared to be closely allied with the euglenozoan CypAs, but constituted an independent lineage. GjCyp-1 showed little relationship with other algal Cyps. A green alga, Chlamydomonas (Chl a + b group), was located in a green plant clade, but a brown alga, Fucus (Chl a + c group), formed an independent clade with a fungus Uromyces (Basidiomycota). PMID- 11911463 TI - Expression of p21WAF1 is dependent on the activation of ERK during vitamin E succinate-induced monocytic differentiation. AB - Vitamin E-succinate (VES) induced monocvtic differentiation of HL-60 human leukemia cells. Treatment with VES increased the nitroblue tetrazolium reduction activity, and the expression of monocyte specific cell surface antigen, CD14 and c-fms. During the monocytic differentiation of HL-60 cells that were induced by VES, the phosphorylation of extracellular signal-regulated protein kinase (ERK) was increased by 12 h and then gradually decreased to a level that was similar to that of the control. However, the phosphorylation levels of p38 and JNK, as well as the expression levels of ERK, p38, and JNK, were unchanged by the VES treatment. Treatment with VES also induced hypophosphorylation of the retinoblastoma protein and an increase of the p21WAF1 protein level. VES-induced ERK phosphorylation was abolished by the ERK inhibitor, PD98059, which resulted in a remarkable prevention of VES-induced monocytic differentiation. Inhibition of the ERK activity by PD98059 also diminished the VES-induced p21WAF1 protein expression, but did not change the phosphorylation state of the retinoblastoma protein. Collectively, these data suggest that the ERK signaling pathway mediates the up-regulation of the p21xWAF1 expression that is induced by VES, which is required for monocytic differentiation of HL 60 cells. PMID- 11911464 TI - Simultaneous expression of allogenic class II MHC and B7.1 (CD80) molecules in A20 B-lymphoma cell line enhances tumor immunogenicity. AB - We expressed the allogenic class II MHC antigen and B7.1 (CD80) co-stimulatory molecule in A20 beta-lymphoma cells in order to test their efficacy as immuno stimulating adjuvant agents in inducing tumor-specific immunity. The transduction of the allogenic I-Ab alpha and beta chain genes into A20 cell resulted in a surface expression of the allogenic class II MHC molecules. The expression of the allogenic class II MHC antigen (I-Ab) in A20 cells enhanced the proliferation of T cells in a mixed lymphocyte tumor culture and in vitro cytotoxic T lymphocyte (CTL) generation against parental cells. The B7.1 gene, which is known to be a potent co-stimulatory molecule, was also transduced and expressed in A20 cells, either alone or in combination with I-Ab. The B7.1 transduction alone leads to a similar in vitro immune enhancing effect as I-Ab. When both the I-Ab and B7.1 genes were transduced, the in vitro immunostimulating capacity was further enhanced. Finally, we also tested the A20 cells that were transduced with I-Ab and/or B7.1 for their efficacy as preventive tumor vaccines in vivo. The results indicate that the A20 cells that express both the I-Ab and B7.1 have more potent vaccinating potential, compared to the cells that express only one of the molecules. PMID- 11911465 TI - Characterization of mycoplasma arginine deiminase expressed in E. coli and its inhibitory regulation of nitric oxide synthesis. AB - We previously reported that a cytostatic protein that is found in ASC-17D Sertoli cell-conditioned media was Mycoplasma arginine deiminase (ADI), which hydrolyzes L-arginine into L-citrulline and ammonia. Here, we report the over-expression of recombinant ADI (rADI) in E. coli and the down-regulation of lipopolysaccharide (LPS) induced-nitric oxide (NO) production by rADI treatment. We cloned the ADI gene from Mycoplasma arginini genomic DNA by a polymerase chain reaction, and changed five TGA tryptophan codons (stop codon in E. coli) to TGG codons in the coding region by site-directed mutagenesis in order to express in E. coli. The rADI was purified to apparent homogeneity by DEAE-Sepharose and arginine-affinity chromatography. The rADI expressed in E. coli was identified as 45 kDa on SDS PAGE and 90 kDa on native PAGE, implying that it exists as a dimer like ADI of M. arginini. The Km for arginine of rADI was approximately 370+/-50 microM. Its optimal temperature and pH were 41 degrees C and pH 6.4, respectively, and enzyme activity remained > or = 50% for 5 d at physiological temperature and pH. Treatment of purified rADI suppressed NO production in macrophage-like RAW 264.7 and primary glial cells that were exposed to LPS. Furthermore, an intraperitoneal injection of rADI significantly suppressed the rise of blood nitrite/nitrate levels that were induced by the systemic administration of bacterial endotoxin LPS to mice, resulting in an improvement in their survival rate. These results suggest that the depletion of blood arginine with an arginine-metabolizing enzyme, such as ADI, could suppress excessive production of NO that is caused by inducible NOS (iNOS) during the endotoxemia. Also, rADI may be used as a new approach to control NO-related diseases, such as sepsis. PMID- 11911466 TI - A leaf-specific 27 kDa protein of potato Kunitz-type proteinase inhibitor is induced in response to abscisic acid, ethylene, methyl jasmonate, and water deficit. AB - The 22 kDa Kunitz-type potato proteinase inhibitor (22 kDa KPPI) was induced in tubers. However, the 27 kDa protein, which is immunologically related to the 22 kDa KPPI, was induced in leaves by wounding, hormones, and environmental stresses. The leaf-specific 27 kDa protein was induced in leaves that were treated with exogenous abscisic acid (ABA), ethephon, methyl jasmonate (MeJA), and water deficit. These results indicate that the 27 kDa protein in leaves could function as a defense protein against mechanical damages by herbivorous animals and abiotic environmental stresses that could induce plant hormones. PMID- 11911467 TI - Development of a new xenoestrogen screening system using fission yeast Schizosaccharomyces pombe. AB - Endocrine disrupters refer to environmental or chemical compounds, which interfere with the endocrine system of organisms. In this study, our aim was to develop a screening method to detect xenoestrogen (an endocrine disrupter that is commonly encountered in our daily life) by using fission yeast Schizosaccharomyces pombe. Although the yeast (the simplest eukaryotic cell) has no endocrine system, estrogen receptors that are created to express in the yeast cell can be activated by estrogen in a similar manner to mammalian cells. First, in order to express the human estrogen receptor (hER) in the yeast cell, we constructed a yeast expression vector that contained hER (pREP42MHN-hER). In the yeast cells that are transformed with the pREP42MHN-hER vector, estrogen receptors could recognize xenoestrogen, which allowed the determination of the presence of xenoestrogen in any given sample. Furthermore, we constructed a yeast strain that contained an estrogen responsive element (ERE) that fused with the Escherichia coli LacZ gene (pERE-LacZ) as a reporter for binding of xenoestrogen with the estrogen receptor. Since this vector system allows determination of the presence and level of xenoestrogen with simple procedures, it is expected that they can serve as an efficient assay system to detect xenoestrogen. PMID- 11911468 TI - Electron paramagnetic resonance study reveals a putative iron-sulfur cluster in human rpS3 protein. AB - The human ribosomal protein S3 (rpS3) functions as a component of the 40S subunit and as a UV DNA repair endonuclease. This enzyme has an endonuclease activity for UV-irradiated and oxidatively damaged DNAs. DNA repair endonucleases recognize a variety of UV and oxidative base damages in DNA from E. coli to human cells. E. coli endonuclease III is especially known to have an iron-sulfur cluster as a co factor. Here, we tried an electron paramagnetic resonance (EPR) method for the first time to observe a known iron-sulfur cluster signal from E. coli endonuclease III that was previously reported. We compared it to the human rpS3 in order to find out whether or not the human protein contains an iron-sulfur cluster. As a result, we succeeded in observing a Fe EPR signal that is apparently from an iron-sulfur cluster in the human rpS3 endonuclease. The EPR signal from the human enzyme, consisting of three major parts, is similar to that from the E. coli enzyme, but it has a distinct extra peak. PMID- 11911469 TI - Functional studies on the interaction between human replication protein A and Xeroderma pigmentosum group A complementing protein (XPA). PMID- 11911470 TI - Production of monoclonal antibodies and immunohistochemical studies of brain myo inositol monophosphate phosphatase. AB - Five monoclonal antibodies that recognize porcine brain myo-inositol monophosphate phosphatase (IMPase) have been selected and designated as mAb IMPP 9, IMPP 10, IMPP 11, IMPP 15, and IMPP 17. These antibodies recognize different epitopes of the enzyme and one of these inhibited the enzyme activity. When the total proteins of the porcine brain homogenate separated by SDS-PAGE were probed with monoclonal antibodies, a single reactive protein band of 29 kDa, co migrating with the purified porcine brain IMPase, was detected. Using the anti IMPase antibodies as probes, the cross reactivities of the brain IMPase from human and other mammalian tissues, as well as from avian sources, were investigated. Among the human and animal tissues tested, the immunoreactive bands on Western blots appeared to have the same molecular mass of 29 kDa. In addition, there was IMPase immunoreactivity in the various neuronal populations in the rat brain. These results indicate that mammalian brains contain only one major type of immunologically similar IMPase, although some properties of the enzymes that were previously reported differ from each another. The first demonstration of the IMPase localization in the brain may also provide useful data for future investigations on the function of this enzyme in relation to various neurological diseases. PMID- 11911471 TI - Cloning of the cel8Y gene from Pectobacterium chrysanthemi PY35 and its comparison to cel genes of soft-rot Pectobacterium. AB - The phytopathogenic Pectobacterium chrysanthemi (Pch) PY35 secretes multiple isozymes of plant cell wall degrading enzyme cellulases. We cloned a second cel gene that encodes cellulase in Pch PY35. The inserted 2 kb fragment was subcloned in order to geneate pPY710 (cel8Y). The structural organization of the cel8Y gene consists of an open reading frame (ORF) of 999 bp that encodes 332 amino acid residues with a signal peptide of 23 amino acids. The predicted amino acid sequence of Cel8Y was very similar to that of Cellulomonas uda, but completely different from that of the Cel5Z of Pch PY35. It belonged to the glycoside hydrolase family 8, based on amino acid sequence similarities in contrast to Cel5Z of Pch PY35, which was confirmed as family 5. Cel8Y was not closely related to the known cellulases of Pectobacterium. It had the conserved region of the glycoside hydrolase family 8, ASDGDVLIAWALLKAGNKW. The apparent molecular mass of the Cel8Y protein was calculated to be approximately 34 kDa by a carboxymethylcellulosesodium dodecyl sulfate-polyacrylamide gel electrophoresis (CMC-SDS-PAGE). The Cel8Y had a calculated pl of 6.49. It was optimally active at pH 7 with an approximate optimal temperature around 40 degrees C. The cellulase activity of Cel8Y was lower than that of Cel5Z. PMID- 11911472 TI - A transcription terminator in the groEx gene of symbiotic X-bacteria expressed in Escherichia coli. AB - The over-expressing groEx gene of symbiotic X-bacteria in Amoeba proteus has unique nucleotide motifs (Tx), containing two hairpins and a C-rich region at its 3'-end. To investigate the role of Tx as a transcription terminator, we mutated Tx and analyzed the effects on the expression of an upstream-located lacZ in E. coli. The level of beta-galactosidase (beta-gal) production in Tx deletion mutants was reduced to 23% of the control level. Site-directed mutation of the hairpin-1, C-rich region, and hairpin-2 reduced the beta-gal production to 28 64%, 33, and 20% of wild-type Tx, respectively. The amount of lacZ transcripts that were detected in RNA blots was proportional to the level of beta-gal. The Tx sequence had 97% termination efficiency in vivo, and the mutations in Tx resulted in read-through transcripts with significantly shortened half-life. In rho- E. coli, the level of the beta-gal production by Tx was reduced to 15% of that in rho+ E. coli. These results suggest that Tx is a Rho-dependent transcription terminator. Also, hairpin-2 is the most effective component among the three motifs of Tx for proper termination of the transcription and stability of mRNAs. PMID- 11911473 TI - Processing and assembly in vitro of engineered soybean beta-conglycinin subunits with the asparagine-glycine proteolytic cleavage site of 11S globulins. AB - A short interdomain sequence between the N- and C-terminal domains of beta conglycinin, the major 7S seed storage protein of soybean, was selected as a target for insertion of amino acid residues specifically cleaved by an asparaginyl endopeptidase that processes globulins into acidic and basic chains. Modified beta-conglycinin subunits containing the proteolytic cleavage site self assembled into trimers in vitro at an efficiency similar to that of the unmodified subunit. In contrast to the absence of cleavage of the unmodified subunits, however, the modified beta-conglycinin trimers were processed by purified soybean asparaginyl endopeptidase into two polypeptides, each the size expected for the beta-conglycinin N- and C-terminal domains, respectively. The cleavage did not alter the assembly of mutant beta-conglycinins and the cleaved mutant trimers remained stable to further proteolytic attack. To examine the possibility of coassembly between the cleaved 11S and 7S subunits, in vitro processed mutant beta-conglycinin subunits were mixed with native dissociated 11S globulin preparations. Reassembly at a high ionic condition did not induce the 7S subunits to interact with 11S subunits to form hexameric complexes. Thus, cleavage of 7S globulin subunits into acidic and basic domains may not be sufficient for hexamer assembly to occur. Biotechnological implications of the engineered proteins are discussed. PMID- 11911475 TI - Generation of infectious cDNA clones of a Korean strain of tomato aspermy virus. AB - Infectious full-length cDNA clones of the Korean strain of tomato aspermy cucumovirus (KC-TAV) were constructed using a long-template reverse transcription polymerase chain reaction. The in vitro RNA transcripts, which were produced using T7 RNA polymerase from full-length cDNAs, could systemically infect the Nicotiana tabacum cv. Xanti-nc plants and induce systemic symptoms on the upper leaves that are similar to the wildtype KC-TAV. The complete nucleotide sequences of genomic RNAs of KC-TAV were determined from the infectious full-length cDNA clones. RNA1 and RNA2 of KC-TAV contain 3412 nucleotides and 3074 nucleotides, respectively. RNA3 of KC-TAV, 2222 nucleotides long, encodes the 3a protein and coat protein (CP) that are separated by 295 nucleotides intergenic region. The overall sequence analysis of the whole genome of KC-TAV revealed a strong homology (99%) to the genome of the V-TAV strain, the only strain whose entire genomic nucleotide sequence was available in the database, and an overall 60% homology to those of other cucumber mosaic virus and peanut stunt virus strains. A sequence comparison analysis of the deduced amino acid sequences of cDNAs of KC TAV RNA 1, 2, and 3 indicates that there is no genetic diversity in the TAV population, although the virus exists in different geographical distributions. PMID- 11911474 TI - Ursolic acid of Origanum majorana L. reduces Abeta-induced oxidative injury. AB - Amyloid beta protein (Abeta) increases free radical production and lipid peroxidation in PC12 nerve cells, leading to apoptosis and cell death. The effect of ursolic acid from Origanum majorana L. on Abeta-induced neurotoxicity was investigated using PC12 cells. Pretreatment with isolated ursolic acid and vitamin E prevented the PC12 cell from reactive oxygen species (ROS) toxicity that is mediated by Abeta. The ursolic acid resulted in decreased Abeta toxicity assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH), and trypan blue assay. Thus, treatment with these antioxidants inhibited the Abeta-induced neurotoxic effect. Therefore, these results indicate that micromolar Abeta-induced oxidative cell death is reduced by ursolic acid from Origanum majorana L. PMID- 11911476 TI - An intronic silencer of the mouse perforin gene. AB - We have previously shown that the perforin gene locus is comprised of eight DNase I hypersensitive sites (DHS). Seven (DHS I-DHS VII) of them were CTL-specific whereas one (DHS VIII) in the second intron was expressed in a wide range of cell types. DHS VIII was highly AT-rich (75%) and was comprised of multiple sets of high mobility group (HMG)-I/Y binding site, two potential Special AT-rich Binding protein (SATB-1)-binding sites, and a long stretch of CTAT repeats, indicating that DHS VIII may relate to nuclear matrix-associated region (MAR). When DHS VIII was inserted into the perforin promoter-driven luciferase gene, it silenced the reporter gene transcription in CTLL-R8 cells in an orientation- and distance independent manner. Moreover, this silencing effect was also observed in other promoters in a variety of non-CTL cell lines, suggesting that DHS VIII exerted a global silencing effect. Deletion analysis and gel-shift assays indicated that the silencing effect was mediated by the CTAT repeats and its binding protein called CTAT repeats-binding protein (CRBP). PMID- 11911477 TI - Purification, characterization, and cDNA cloning of rice class III chitinase. AB - Several chitinases were expressed in a rice cell suspension culture and detected in the medium. One of them, designated as RCB4, was isolated 248 fold from the culture filtrate to homogeneity by 70% ammonium sulfate precipitation, DEAE cellulose, CM-cellulose, Sephadex G-75 column chromatography, and native gel slicing. RCB4 had a molecular mass of 32 kDa by SDS-PAGE. The optimum temperature was 40 degrees C, and 96% of its activity still remained at 60 degrees C. The optimum pH was 4, and 95% of its activity was maintained at pH 2. Using a substrate (GlcNAc)6, the Km and Vmax values of RCB4 were 0.53 mM and 11.1 mM/min, respectively. The N-terminal and internal amino acid sequences of RCB4 were determined to be VNSNLFRDYIGA and MALWA, respectively. A cDNA (C12523) clone that contained the N-terminal and internal amino acid sequences of RCB4 was obtained, sequenced, and renamed RCB41. RCB41 encoded 307 amino acid protein with a signal peptide of 25 amino acids and showed a 45% similarity to gladiolus chitinase GBC a, one of the class III chitinase family. The expression of RCB4l in E. coli showed that RCB41 encodes a chitinase. PMID- 11911478 TI - Molecular mechanism of NFAT family proteins for differential regulation of the IL 2 and TNF-alpha promoters. AB - Nuclear factor of activated T cells (NFAT) is a family of transcription factors that regulates activation-induced transcription of many immunologically important genes. Although all NFAT family proteins contain a highly conserved DNA-binding domain and also bind cooperatively with AP-1 proteins to the interleukin-2 (IL-2) promoter NFAT site, each member shows characteristic site preferences to other promoters. Previously, we have shown that NFATc.beta, an isoform of NFATc, is different from NFATp in both DNA binding and transactivation of the TNF-alpha promoter. To further characterize target gene specificity of NFATc and NFATp, we generated deletion mutants as well as mutants swapping the C-terminal region of their DNA binding domains, and analyzed their DNA-binding specificity to different target sites. Our results show that the C-terminal one third of DNA binding domain confers different binding specificity of NFATc and NFATp to an NFAT site in the TNF-alpha promoters. Transient expression of the mutant NFAT proteins also demonstrates that transcriptional activation of the target promoters is consistent with the DNA binding specificity of the mutant NFATs. These results strongly suggest that a binding site preference and availability of different NFAT proteins may program the temporal expression of distinct cytokine genes. Importantly, the C-terminal region of the DNA binding domain plays an important role in determining the binding site preferences at least of NFATp and NFATc members. PMID- 11911479 TI - Tcp10 promoter-directed expression of the Nek2 gene in mouse meiotic spermatocytes. AB - Nek2 is a cell cycle regulator that is involved in diverse cell cycle events. The expression pattern and biochemical properties of Nek2 in mammalian male germ cells suggested its involvement on regulation of the meiotic cell cycle. To further investigate specific roles of Nek2 in meiosis, we generated transgenic mice in which the Nek2 transgenes were expressed specifically in spermatocytes using the Tcp10 promoter. The Nek2 transgenic mice did not reveal any significant defect in gross testicular anatomy as well as in fertility. However, we observed significant increases in defective spermatogenic cells, such as apoptotic cells and giant degenerating cells, in the Tcp10/Nek2 transgenic mice. These results revealed that even only slightly elevated production of the Nek2 protein disturbed the normal development of male germ cells, possibly in meiosis. PMID- 11911480 TI - Characterization of lipopolysaccharide responsive proteins that binds to the 5' flanking regions of mouse Rantes. AB - When macrophage (like the RAW264.7 cell line) was stimulated with lipopolysaccharide (LPS), factors that bind specifically to the LPS responsive element (LRE) of murine Rantes gene appeared in the nucleus. An electrophoretic mobility shift assay (EMSA) detected 2 specific bands, designated as S (slow) and M (middle). The S band appeared within 15 min of LPS stimulation, and reached its highest intensity within 2 h. The M band was present in unstimulated cells, but after stimulation its intensity increased and reached its highest intensity also in about 2 h. Significantly, in LPS hyporesponsive 10-9 macrophage like cells, the S band was absent. The M band was present in equal amounts in stimulated and unstimulated cells. The results suggest that the S band was induced by LPS stimulation. In the nuclear extract, the native molecular weight of the S band forming factor was approximately 270 kDa, and that of the M bands-forming factor was approximately 140 kDa. U.V. cross linking studies consistently showed at least 2 different polypeptides of approximate molecular mass of 70 kDa, both in the S band-forming factor (complex) and the M band-forming factor (complex). In the nuclear extracts of both the LPS stimulated and unstimulated cells, we detected a factor with approximate molecular mass of 120 kDa that could convert the S band-forming complex to the M band-forming complex. This factor, designated as a converting factor, is a protein phosphatase since its activity was blocked by okadaic acid, an inhibitor of Ser/Thr protein phosphatase. Also, purified protein phosphatase type 1 (PP-1) could convert the S band-forming complex to the M band-forming complex. PMID- 11911481 TI - Molecular cloning and characterization of cDNAs encoding heterotrimeric G protein alpha and beta subunits from potato (Solanum tuberosum L.). AB - Two cDNAs, STGA2 and STGB2, that encode heterotrimeric G protein alpha and beta subunit proteins, respectively, were cloned from an early tuber cDNA library of potato (Solanum tuberosum cv. Superior). The cDNA of STGA2 encoded 384 amino acids, which showed 75-98% identities to plant Ga-subunits; STGB2 encoded 377 amino acids, which showed 83-92% identities to plant Gbeta-subunits. The transcript levels of the two genes were abundant in leaves, shoots, axially buds, unopened flowers, and active growing sprouts. However, the transcripts were barely detectable in roots. The expressions of STGA2 and STGB2 were up-regulated by light. Interestingly, the STGA2 and STGB2 gene expression showed synchronous patterns in the examined organs. During the early tuber development, the transcripts of STGA2 and STGB2 were abundant in unswollen stolons, swollen stolons, and new tubers, but were undetected in matured tubers. This indicates that potato Galpha- and beta-subunit genes are developmentally regulated. Based on these observations, we propose that heterotrimeric G proteins may be involved in the signaling pathway during potato tuber development. PMID- 11911482 TI - Treatment of experimental rabbit liver tumours by selectively targeted hyperthermia. AB - Experimental rabbit liver tumours were preferentially heated to therapeutic temperatures without compromising the surrounding normal hepatic parenchyma. This was achieved by the use of hepatic arterially infused ferromagnetic microspheres that heat as a result of magnetic hysteresis loss when exposed to an alternating magnetic field. Treatment sessions involving a single 20-min exposure to the alternating field resulted in total suppression of tumour growth at 14 days compared to controls, in which tumour sizes increased dramatically over the same period. Histopathological examination of treated tumour sections showed total tumour destruction in some cases. Separate animal groups used to control for the effects of the embolized microspheres alone and for the effect of the applied magnetic field yielded similar tumour growth responses to a control group with no intervention whatsoever. The achievement of positive temperature differentials between tumour and normal liver and the consequent therapeutic responses encourages further development of this technology for the treatment of liver cancer in humans. PMID- 11911483 TI - The effect of tumour size on ferromagnetic embolization hyperthermia in a rabbit liver tumour model. AB - OBJECTIVE: Ferromagnetic Embolization Hyperthermia (FEH) consists of arterially embolizing tumours with ferromagnetic particles to cause hysteretic heating upon subsequent exposure to an alternating magnetic field. The objective was to determine the effect of tumour size during FEH using a rabbit liver tumour model. METHOD: Thirty-three rabbits containing implanted hepatic VX2 carcinomas received a hepatic arterial infusion of ferromagnetic particles suspended in lipiodol. Following hysteretic heating, tumour and normal hepatic tissues were chemically analysed for iron content. Tumours were classed as small if their mass was less than the median mass for the whole group of subjects (2.1 g), and as large if their mass was greater than or equal to the median. To control for variability in tumour iron concentration, 13 small tumours were matched to 13 large tumours by iron concentration, and their heating characteristics compared. RESULTS: The heating rate in large tumours (median = 5.0 degrees C/min) was significantly greater than that in the matched small tumours (median = 2.8 degrees C/min), p = 0.006. Regression analysis determined that the slope of the heating rate vs iron concentration curve for large tumours was 1.5 times greater than that for the matched small tumours, p < 0.001. After cessation of heating in large tumours, there was continued heat dissipation into surrounding tissues, which led to anomalous temperature increases. There was an inverse linear relationship between tumour size and tumour iron concentration for a given dose of particles. CONCLUSION: For a given tumour iron concentration, larger tumours heat at a greater rate than small tumours, due to the poorer tissue cooling and better heat conduction in the necrotic regions of large tumours. This warrants further investigation as this finding could confer a significant advantage on FEH over other hyperthermic modalities in the treatment of hepatic malignancies. PMID- 11911484 TI - Hyperthermia exhibits anti-vascular activity in the s.c. BT4An rat glioma: lack of interaction with the angiogenesis inhibitor batimastat. AB - We investigated the anti-vascular activity of local hyperthermia (44 degrees C, 60 min) in s.c. BT4An rat gliomas, and the influence on tumour growth of hyperthermia and the anti-angiogenic compound batimastat (30 mg/kg i.p.). Heat induced vascular damage was assessed in small (82 mm3) and large (171 mm3) tumours using confocal microscopy and immunostaining for von Willebrand factor. The influence of hyperthermia on tumour blood flow was measured using the 86RbCl method. The anti-tumour activity of hyperthermia (one or two fractions a week apart) and batimastat (7 or 14 daily injections) and various combination schedules were examined. Hyperthermia disrupted 25-50% of the vasculature in the BT4An tumours, and the vascular damage was most extensive in the central part of the large tumours. The heated tumours exhibited a 40-60% blood flow reduction, which persisted until the last measurement after 24 h. One fraction of hyperthermia caused a significant growth delay of 4 days, compared with the control group (p = 0.01), but no additional tumour response was produced by a second heating session. Batimastat had no influence on tumour growth and the combination of drug and local heating did not enhance the tumour response, compared with heating alone. It was concluded that hyperthermia at 44 degrees C for 60min exhibits anti-vascular activity and inhibits tumour growth in the BT4An tumour model. Batimastat had no effect on tumour growth. PMID- 11911485 TI - Betulinic acid sensitization of low pH adapted human melanoma cells to hyperthermia. AB - Betulinic acid is a known inducer of apoptosis in human melanoma that is most effective under conditions of low pH. It was hypothesized that betulinic acid, in combination with acute acidification and/or hyperthermia, would induce higher levels of apoptosis and cytotoxicity in low pH-adapted human melanoma cells than in cells grown at pH 7.3. DB-1 human melanoma cells, adapted to a tumour-like growth pH of 6.7, were exposed to hyperthermia (2h at 42 degrees C) and/or betulinic acid (4-10 microg/ml) and compared with cells grown at a physiological pH of 7.3 or after acute acidification from pH 7.3-6.3 or pH 6.7-6.3. Betulinic acid induced higher levels of apoptosis and cytotoxicity in low pH-adapted cells than in cells grown at pH 7.3, as measured by the terminal deoxynucleotidyl transferase (TdT) DNA fragmentation assay (TUNEL), the MTS cell viability assay, and single cell survival. Acute acidification of low pH adapted cells rendered them more susceptible to betulinic acid-induced apoptosis and cytotoxicity. In the presence of hyperthermia at 42 degrees C for 2 h, cells grown at pH 7.3 were not sensitized to heat killing by betulinic acid, whereas cells grown at pH 7.3 and acutely acidified to pH 6.3, cells adapted to growth at pH 6.7 and cells adapted to growth at pH 6.7 and acutely acidified to pH 6.3 were all similarly sensitized to heat killing by betulinic acid, with survival values of 5, 9 and 2%, respectively. It is concluded that betulinic acid may be useful in potentiating the therapeutic efficacy of hyperthermia as a cytotoxic agent in acidotic areas of tumours with minimal effect in normal tissues growing at pH 7.3. PMID- 11911486 TI - Non-invasive temperature profile estimation in a lossy medium based on multi-band radiometric signals sensed by a microwave dual-purpose body-contacting antenna. AB - Microwave radiometry has during the past two decades been investigated as a non invasive scheme for measurement of subcutaneous tissue temperatures, basically for monitoring and control in hyperthermic treatment of cancer. In this effort, we test a contact-type, dual-purpose antenna with integral water bolus. To overcome conflicting optimization criteria in the integration of this thermometry technique with heat applicators exhibiting a large effective field size during superficial hyperthermia, a stacked configuration design is proposed, where the radiometer receive antenna (Archimedean spiral) is located on the front (skin) surface of the water bolus and the heating antenna (Dual-Concentric Conductor aperture) is placed on the bolus back surface. The motivation is to achieve homogeneous tissue heating simultaneously with non-invasive thermography of the target tissue under the applicator. This paper addresses the feasibility of predicting one-dimensional depth temperature profiles from multi-band brightness temperatures. The performance is investigated statistically by a Monte Carlo technique on both simulated and real heated-phantom data using up to six radiometric bands. Analysis of measured data shows that during the transient heating period in a solid lossy phantom, the inversion technique exhibits a precision (2sigmaT) and skewness (bias) of estimated compared to actual temperature profiles of better than +/-0.38 degrees C and +/-0.55 degrees C, respectively. PMID- 11911487 TI - Health risk assessment on residents exposed to chlorinated hydrocarbons contaminated in groundwater of a hazardous waste site. AB - We conducted this study to estimate residents' chronic hazard and carcinogenic risk in a groundwater-contaminated community after on-site remediation in Taiwan during 1999-2000. We followed guidelines for assessing hazardous waste sites of the U.S. Environmental Protection Agency (EPA) and used empirically measured contaminant levels and exposure parameters to perform health risk assessment on seven chlorinated hydrocarbons. We measured groundwater concentrations of vinyl chloride, tetrachloroethylene, trichloroethylene, 1,1-dichloroethylene, 1,1,1 trichloroethane, cis-1,2-dichloroethylene, and 1,1-dichloroethane in 49 off-site residential wells by gas chromatography/mass spectrometry. Exposure parameters were mainly derived from our field survey of 382 residents, and partially from U.S. EPA default values. Total exposure dose estimation included routes of inhalation during showering and dermal absorption of showers and other activities involved with hand-water contacts. The ingestion route of water was not included because most residents drank boiled water with negligible contaminants. We calculated a hazard index (HI) for all seven chlorinated hydrocarbons and carcinogenic risks for known human carcinogen of vinyl chloride and probable human carcinogens of tetrachloroethylene and trichloroethylene, which had the same target organ, the liver. The HI values for reasonable maximal exposure (RME) and average exposure were 14.3 and 0.2, respectively. The cancer risks based on RME and average exposure (in parentheses) for vinyl chloride, tetrachloroethylene, and trichloroethylene were 8.4 x 10(-6) (7.3 x 10(-9)), 1.9 x 10(-4) (1.3 x 10(-7)), and 1.4 x 10(-4) (1.2 x 10(-6)), respectively. We applied Monte Carlo simulations to the sensitivity analysis, which showed that the contaminant levels, exposure duration, and time for showers were major determinants of health risks. We concluded that the contaminated groundwater was still unsafe for use even after the contaminated site underwent remediation by extraction and treatment in 1997. PMID- 11911488 TI - Respiratory and irritant health effects in tollbooth collectors in Taiwan. AB - Toll collectors are potentially exposed to high concentrations of a variety of automotive emissions. The purpose of this study is to assess if there is an excess of adverse health outcomes among toll collectors. Self-reported chronic respiratory symptoms and acute irritative symptoms were assessed in a cross sectional study among 363 toll collectors (exposure group) and 147 office workers (control group). The prevalence rates for chronic respiratory symptoms were not significantly different between the exposure group and the control group. This could be related to the short length of employment for the toll collectors. Another likely explanation for not detecting apparent respiratory effects might be that symptomatic workers drop out of this work a few years after starting the job. Acute irritative symptoms (nose, throat, nausea, and headache), however, were significantly more common among the toll collectors than among the controls. PMID- 11911489 TI - Arsenite pretreatment attenuates benzo[a]pyrene cytotoxicity in a human lung adenocarcinoma cell line by decreasing cyclooxygenase-2 levels. AB - Both simultaneous and sequential exposure to arsenite and benzo[a]pyrene (BaP) potentially occur in human populations drinking arsenic-contaminated water or burning arsenic-contaminated coal. Although arsenite and BaP are both well documented hazardous substances and human carcinogens, interactions between these two agents have not been well defined. In this study, we demonstrated that posttreatment with arsenite synergistically enhanced the cytotoxicity of BaP for a human lung adenocarcinoma cell line, CL3. In contrast, pretreatment of CL3 cells with arsenite attenuated BaP cytotoxicity. Involvement of heat-shock protein 70 and heme oxygenase-1 in this arsenite-mediated attenuation of BaP cytotoxicity was ruled out. Our data also indicated that arsenite pretreatment did not affect the BaP-mediated induction of CYP1A1, the initial enzyme involved in its metabolic activation, but did result in a significant decrease in mRNA and protein levels of cyclooxygenase-2 (COX-2), which is required to convert the BaP metabolite BaP 7,8-dihydrodiol to the ultimate epoxide. In contrast to the high susceptibility of CL3 cells to BaP, the human lung carcinoma cells, H460, and CL3R15 cells (arsenic-resistant CL3 cells) showed normal CYP1A1 inducibility by BaP, had negligible amounts of COX-2, and were highly resistant to BaP. The involvement of COX-2 in BaP activation was confirmed by transfection of H460 cells with a recombinant adenovirus, Ad-pgk-Cox2, coding for COX-2, which resulted in a significant increase in the levels of the COX-2 product prostaglandin E2 in the medium and in the susceptibility of H460 cells to BaP. The present study confirms the importance of COX-2 in BaP activation and demonstrates that the arsenite-mediated attenuation of BaP cytotoxicity is mediated by a reduction in COX-2 levels. PMID- 11911490 TI - Human 8-oxoguanine DNA glycosylase 1 mRNA expression as an oxidative stress exposure biomarker of cooking oil fumes. AB - Epidemiological studies have indicated that the exposure to carcinogenic components formed during the cooking of food might be associated with lung cancer risk of Chinese women. Previous studies have confirmed that cooking oil fumes from frying fish (COF) contained relatively high amount of benzo[a]pyrene, 2 methyl-3,8-dimethylimidazo[4,5-f] qunoxaline, benzene, and 1,3-butadiene, reported in fumes from heated soybean oil. Thus, we consider that oxidative stress induced by COF may play a role in lung cancer development among Chinese women. To verify whether the oxidative DNA damage was induced by COF, high performance liquid chromatography (HPLC) analysis data showed that the levels of 8-hydroxydeoxyguanine (8-OH dG) were increased in a dose-dependent manner when calf thymus DNA reacted with various concentrations of COF. Since human 8 oxoguanine DNA glycosylase 1 (hOGG1) was a repair enzyme for removing 8- OH dG from damaged DNA, we hypothesized that hOGG1 mRNA may be used to assess the risk of oxidative damage induced by the exposure of COF. The results from reverse transcription polymerase chain reaction showed that the hOGG1 mRNA expression was induced by hydrogen peroxide (H2O2) and COF in human lung adenocarcinoma CL-3 cells. To elucidate whether hOGG1 mRNA expression was an exposure biomarker of COF, a cross-sectional study of 238 subjects including 94 professional cooks, 43 housewives, and 101 COF-nonexposed control subjects was conducted. The hOGG1 mRNA expression frequencies of COF-exposed cooks (27 of 94, 28.7%) and housewives (6 of 43, 14%) were significantly higher than those of control subjects (4 of 101, 4%). After adjusting for age, sex, and smoking and drinking status, the odds risks (ORs) of housewives versus control and cooks versus control were 3.94 (95% confidence interval [CI] = 0.95-16.62) and 10.12 (95% CI = 2.83-36.15), respectively. These results indicated that hOGG1 may be adequate to act as an exposure biomarker to assess the oxidative DNA damage induced by COF. This also suggests that oxidative stress induced by COF may play a role in lung cancer development among Chinese women. PMID- 11911491 TI - Chronic toxicity of a mixture of chlorinated alkanes and alkenes in ICR mice. AB - The aim of this study was to determine the chronic toxicity of a mixture of chlorinated alkanes and alkenes (CA) consisting of chloroform, 1,1 dichloroethane, 1,1-dichloroethylene, 1,1,1-trichloroethane, trichloroethylene, and tetrachloroethylene. These chlorinated organic solvents were present in the underground water near an electronic appliances manufactory in Taoyuan, Taiwan. Male and female weanling ICR mice were treated with low-, medium-, and high-dose CA mixtures in drinking water for 16 and 18 mo, respectively. A significant number of male mice treated with the high-dose CA mixture developed tail alopecia and deformation, which was not prominent in CA-treated female mice. Medium- and high-dose CA mixtures induced marginal increases of liver and lung weights, blood urea nitrogen, and serum creatinine levels in male mice. In female mice, the high dose CA mixture increased liver, kidney, and uterus and ovary total weights, without affecting serum biochemistry parameters. CA mixtures had no effects on the total glutathione content or the level of glutathione S-transferase activity in the livers and kid- neys of male and female mice. Treatments with CA mixtures produced a trend of increasing frequency of hepatocelluar neoplasms in male mice, compared to male and female controls and CA-treated female mice. The high-dose CA mixture induced a significantly higher incidence of mammary adenocarcinoma in female mice. The calculated odds ratios of mammary adenocarcinoma in female mice induced by low-, medium-, and high-dose CA mixtures were 1.14, 1.37, and 3.53 times that of the controls, respectively. The low-dose CA mixture induced a higher incidence of cysts and inflammation in and around the ovaries. This study has demonstrated that the CA mixture is a potential carcinogen to male and female mice. These animal toxicology data may be important in assessing the health effects of individuals exposed to the CA mixture. PMID- 11911492 TI - Aliphatic chlorinated hydrocarbons alter the contractile responses of tracheal smooth muscle in piglet. AB - Aliphatic chlorinated hydrocarbons (ACHs) are widely used in several industrial processes and are also found in many commercial household products. They are classified as hazardous air pollutants, since ACHs exposure induces respiratory complications including airway hyperactivity. However, the contribution of airway smooth muscle tone to ACH-induced respiratory dysfunction has not been elucidated. Thus, the effects of ACHs such as dichloromethane (DCM), dichloroethane (DCE), and trichloromethane (TCM), on the basal and stimulant induced contractile responses in piglet tracheal smooth muscle were investigated. ACHs at 100-1,000 ppm were found to evoke the basal contraction of tracheal smooth muscle strips. Although DCM, DCE, and TCM enhanced the muscle tone precontracted by KCl, they exerted differential effects on acetylcholine- or histamine-induced muscle contraction. DCE did not alter the muscle tone activated by acetylcholine and histamine. DCM at 1,000 ppm enhanced the muscle tension precontracted by acetylcholine but not by histamine. TCM at 30-1,000 ppm increased the histamine-induced muscle contraction, but at 1,000 ppm relaxed the muscle precontracted by acetylcholine. DCE and TCM at the highest concentration (1,000 ppm) provoked a biphasic response with an initial increase in KCI-induced muscle tension followed by a decrease. Furthermore, pretreatment with DCE potentiated the acetylcholine-, histamine-, and KCl-induced muscle contractile responses. Pretreatment with TCM potentiated the histamine-, and KCl-induced response, but DCM only potentiated the KCl-induced response. The results suggest that ACH exposure altering the basal and spasmogen-induced contractile responses might participate in airway impairment with hyperresponsiveness. PMID- 11911493 TI - Brain lipid peroxidation and changes of trace metals in rats following chronic manganese chloride exposure. AB - The aim of this study was to investigate the effects of chronic, daily, 30-d administration of manganese chloride (MnCl2) to male Sprague-Dawley rats on lipid peroxidation and changes of trace elements (manganese, iron, copper, zinc) in various brain regions. Rats were intraperitoneally injected with MnCl2 (20 mg/kg) once daily for 30 consecutive days. The Mn accumulated in frontal cortex, corpus callosum, hippocampus, striatum, hypothalamus, medulla, cerebellum, and spinal cord. Malondialdehyde, an end product of lipid peroxidation, was markedly decreased in frontal cortex and cerebellum. An increased level of Cu was observed in frontal cortex, medulla, and a cerebellum. A decreased Fe level was found only in cerebellum, and a decreased Zn level was observed in hippocampus and striatum. In a second group of animals, Mn (20 mg/kg/d) and glutathione (GSH, 15 mg/kg/d) were administered ip for 30 d. In CSH-Mn-treated rats, compared to Mn-treated rats, MDA concentrations were significantly reduced in frontal cortex, medulla and cerebellum. The changes of trace elements in rat brain were similar to the Mn treated group. We suggest that Mn is an atypical antioxidant, as well as not involved in oxidative damage in rat brain. Fe and Cu may play roles in the protective effect of Mn against lipid peroxidation in rat brain. PMID- 11911494 TI - Mercuric chloride alters the membrane potential and intracellular calcium level in mouse pancreatic islet cells. AB - In this study, mercuric chloride was applied to the primary cultures of mouse pancreatic islet cells for studying its effects on resting membrane potential and the intracellular free calcium ion concentration ([Ca 2+), using the techniques of electrophysiology and fluorometry. It was observed that mercuric chloride (1 100 microM) caused a rapid and sustained depolarization, and induced a rapid first phase and a large sustained second phase of elevation in fura-2 fluorescence ratio in islet cells. The depolarization and increased lCa2+]i induced by mercuric chloride could be inhibited by dithiothreitol (a sulfhydryl containing reducing agent). Removing Ca2+ from the external medium inhibited the mercuric chloride-induced elevation of [Ca2+]i. The increased [Ca2+]i may also originate from the endoplasmic reticulum of pancreatic islet cells, since caffeine (an activator of Ca2+ release from endoplasmic reticulum) and thapsigargin (an inhibitor of endoplasmic reticulum Ca2+-ATPase) could antagonize the effect of mercuric chloride. Moreover, in the absence of glucose in the medium, the response of islet cells to mercuric chloride was a rapid first phase of increased [Ca2+]i followed by a small sustained second phase. Readministration of 5 mM glucose was sufficient but transient to restore sustained phase of increased [Ca2+]i. The increase of [Ca2+]i in islet cells induced by a lower concentration of mercuric chloride (5 microM) was potentiated in higher glucose (7.5 mM) medium. Tolbutamide, an inhibitor of the ATP-sensitive K+-channel, could also inhibit the effect of mercuric chloride. These findings suggest that mercuric chloride initially interacts with the sulfhydryl groups of membrane bound proteins, which may be an ATP-sensitive K+ channel, to cause depolarization of the islet cells. This depolarization triggers Ca2+ influx and then the release of Ca2+ from the endoplasmic reticulum. PMID- 11911495 TI - Hydroxyl radical formation and oxidative DNA damage induced by areca quid in vivo. AB - Chewing areca quid (AQ) has been implicated as a major risk factor for the development of oral squamous-cell carcinoma (OSCC). Recent studies have suggested that AQ-generated reactive oxygen species (ROS) is one of the contributing factors for oral carcinogenesis. However, the AQ used in Taiwan is different from that used in other countries. This study is designed to test whether ROS are generated and the consequent effects in locally prepared AQ in vivo. We measured the hydroxyl radical formation, as represented by the presence of o- and m tyrosine in saliva from volunteers who chewed AQ containing 20 mg phenylalanine. Their saliva contained significantly higher amounts (p < .05) of o- and m tyrosine as compared to the controls. In addition, chewing AQ containing Piper betle inflorescence generated higher amounts of m-tyrosine, but not o-tyrosine, in saliva than did chewing AQ containing betel leaf. We further tested the oxidative DNA damaging effect of the reconstituted AQ, as evidenced by the elevation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) levels, in hamster buccal pouch. Following daily painting for 14 d, the 8-OH-dG level in hamster buccal pouch is significantly elevated (p < .05) in the AQ-treated group versus the controls. These findings demonstrate that ROS, such as hydroxyl radical, are formed in the human oral cavity during AQ chewing, and chewing such prepared AQ might cause oxidative DNA damage to the surrounding tissues. PMID- 11911496 TI - Regulation of cerebral cortical blood flow by the basal forebrain cholinergic fibers and aging. AB - This article reviews the study of neural vasodilator mechanisms of the cerebral cortex by basal forebrain cholinergic nerve fibers and their age-related function in rats. During the last decade, we have demonstrated a neural regulatory system of cerebral blood flow in rats involving intracerebral cholinergic vasodilator nerve fibers originating in the basal forebrain and projecting to the cerebral cortex. Activation of these cholinergic vasodilator fibers results in the release of acetylcholine (ACh) within the cortex, activation of both nicotinic and muscarinic ACh receptors, and vasodilatation without coupling to glucose metabolic rates. This cholinergic vasodilator system has been shown to decline with age in rats mainly due to age-related declines of nicotinic ACh receptor activity. However, muscarinic ACh receptor activity and the release of ACh into the extracellular space in the cortex are well maintained during aging. The present age-related decline of the intracerebral cholinergic vasodilator system found in rats seems to affect cognitive function during aging, although this cholinergic vasodilator system has not yet been demonstrated in humans. PMID- 11911497 TI - Aging and calcium buffering in adrenergic neurons. AB - The aging process at the cellular, organ and whole organism levels is in many respects a mystery. A common bias among those who study aging is that cellular homeostasis "generally falls apart". The assumption of a general deterioration in cellular homeostasis does not take into account that many individuals age quite well maintaining even robust physiological and mental functions. One facet of aging studies that has come to the forefront is the impact of age on the control of the ion messenger, calcium. Emerging evidence suggests that despite age related declines in any one component or multiple components of the calcium buffering systems, compensatory mechanisms may be able to maintain overall calcium homeostasis. This brief review focuses specifically on the ability of peripheral neurons to maintain control of the ion messenger calcium with advancing age. In addition, the idea that the impact of age on calcium homeostasis may be more subtle due to complex and integrated mechanisms that control this ion is discussed. PMID- 11911498 TI - Selective vulnerability in adult and ageing mammalian neurons. AB - Data are presented in support of the idea of antagonistic pleiotropy that features which are adaptive during early life may become maladaptive during the ageing process, when selective pressure is reduced. A model of selective vulnerability to age-related neurodegeneration involving neighbouring subpopulations of vulnerable and protected sympathetic neurons is presented. The two groups of neurons are morphologically and physiologically distinct, indicating advantageous adaptation to particular functions. Neurotrophin signalling is investigated in these different groups of neurons, revealing significant differences between them: neurotrophic factor expression in their target tissues is markedly different, same as their neurotrophin uptake characteristics. Preliminary evidence is presented that the mechanism linking neurotrophin signalling and age-related neurodegeneration may involve the capacity of neurons to buffer free radical generation, hence reducing the effects of attrition by free radical damage. PMID- 11911499 TI - Aging and neuronal plasticity: lessons from a model. AB - In spite of many well-documented examples of age-related reductions in neuronal plasticity, the causes of such changes remain largely unknown. One example of age reduced plasticity involves an aberrant sprouting response of mature rat sympathetic neurons into the CNS (hippocampal formation). This phenomenon has proven to be useful for exploring the relative contribution of target aging (extrinsic influences) versus neuronal aging (intrinsic influences) to reduced sprouting. Aged sympathetic neurons mount a robust growth response when confronted with young target tissue or when exposed to exogenous trophic factor in vivo. In contrast, the aged target tissue (the hippocampal formation in this example) exhibits reduced receptivity for sympathetic sprouting. This change in the target does not appear to be due to alterations in baseline levels of trophic or substrate support for axonal growth. Rather, aging appears to dampen the consequences of target denervation so that the aged target elicits less sprouting. Age-related reductions in neuronal sprouting are speculated to reflect increasing commitment to information storage at the expense of neuronal plasticity. PMID- 11911500 TI - Modulation of parasympathetic neuron phenotype and function by sympathetic innervation. AB - Selective sympathetic nerve dysfunction occurs during aging and in certain disease states. Here, we review findings concerning the effects of chronic sympathetic denervation on parasympathetic innervation to orbital target tissues in the adult rat. Long-term sympathetic denervation was induced by excising the ipsilateral superior cervical ganglion for 5-6 weeks prior to analyses. Following sympathectomy, pterygopalatine ganglion parasympathetic neurons show reduced nitric oxide synthase protein in their somata and projections to vascular targets. Laser Doppler measurements of ocular blood flow indicate that sympathectomy is also accompanied by reduced nitrergic vasodilatation. In the superior tarsal muscle of the eyelid, parasympathetic varicosities, normally, are distant to smooth muscle cells but make axo-axonal contacts with sympathetic nerves, consistent with physiological evidence showing only prejunctional inhibitory effects on sympathetically mediated smooth muscle contraction. Following sympathectomy, parasympathetic varicosities proliferate and closely appose smooth muscle cells, and this is accompanied by establishment of parasympathetic-smooth muscle excitatory neurotransmission. Many pterygopalatine parasympathetic neurons normally contain nerve growth factor (NGF) protein and express NGF mRNA. However, following chronic sympathectomy or elimination of sympathetic impulse activity, NGF mRNA and protein are markedly reduced, indicating that sympathetic neurotransmission enhances NGF expression in parasympathetic neurons. Together, these findings portray a striking dependency of parasympathetic neurons on sympathetic nerves to maintain normal phenotype and function. Sympathetic influences on parasympathetic neurons may be mediated, in part, through axo-axonal synapses. NGF synthesis and release by parasympathetic neurons may represent a molecular basis underlying the formation of these synapses, and up-regulation of NGF synthesis by sympathetic nerve activity may act to reinforce these associations. PMID- 11911501 TI - Impairment of peripheral sensory innervation in senescence. AB - Sensorimotor disturbances are common among elderly and one of the main factors depreciating life quality in senescence. Mechanistically sensory deficits during aging include not only degenerative and regressive events but also phenotypic switches among sensory neurons as well as remodeling of sensory innervation. The pattern of changes suggests that an underlying mechanism is a sustained dependence of sensory neurons on target tissues, and that this dependence, at least in part, appears to be mediated through signaling by target-derived trophic factors. This review presents and discusses evidence supporting this notion. PMID- 11911502 TI - Neurodegeneration, parkinsonian syndromes and autonomic failure. PMID- 11911503 TI - New findings on the neuropathology of multiple system atrophy. AB - Multiple system atrophy (MSA) provides a typical example of the integrative role of the central autonomic network in controlling cardiovascular, respiratory, bladder and gastrointestinal functions. There is increasing evidence that neurochemically defined neuronal groups of the brainstem are selectively affected in MSA to a much greater degree than in Parkinson's disease. These include the catecholaminergic neurons of the rostral ventrolateral medulla (C1 group) which project to the intermediolateral cell column and are involved in modulation of sympathetic vasomotor outflow, and noradrenergic neurons of the caudal ventrolateral medulla (A1 group) projecting to the magnocellular nuclei of the hypothalamus and regulating vasopressin (AVP) release. Loss of these groups of neurons may, at least in part, explain the development of orthostatic hypotension, baroreflex dysfunction, and impaired reflex AVP release in response to hypotension. There is preliminary evidence that cardiovagal neurons of the ventrolateral portion of the nucleus ambiguus, distinct from the branchimotor neurons of the compact region, may also be affected in MSA. Loss of cholinergic neurons in the medullary arcuate nucleus, considered by some to be the homologous to the central chemosensitive region of the ventral medullary surface, may contribute to disturbances in automatic ventilation, particularly during sleep, in patients with MSA. PMID- 11911504 TI - Age-related sympathetic ganglionic neuropathology: human pathology and animal models. AB - Systematic studies of the autonomic nervous system of human subjects and development of well-defined animal models have begun to substantially improve our understanding of the pathogenesis of autonomic dysfunction in aging and may eventually provide strategies for intervention. Neuropathological studies of the sympathetic ganglia of aged human subjects and rodent models have demonstrated that neuroaxonal dystrophy involving intraganglionic terminal axons and synapses is a robust, unequivocal and consistent neuropathological finding in the aged sympathetic nervous system of man and animals. Quantitative studies have demonstrated that markedly swollen argyrophilic dystrophic axon terminals develop in the prevertebral superior mesenteric (SMG) and coeliac, but to a much lesser degree in the superior cervical ganglia (SCG) as a function of age, sex (males more than females) and diabetes. Dystrophic axons were immunoreactive for neuropeptide Y, tyrosine hydroxylase, dopamine-beta-hydroxylase, trkA and p75NTR, an immunophenotype consistent with their origin from postganglionic sympathetic neurons, and contained large numbers of highly phosphorylated neurofilaments or tubulovesicular elements. The sympathetic ganglia of aged rodents also showed the hallmark changes of neuroaxonal dystrophy as a function of age and location (many more in the SMG than in the SCG). Plasticity-related synaptic remodeling could represent a highly vulnerable target of the aging process. The fidelity of animal models to the neuropathology of aged humans suggests that similar pathogenetic mechanisms may be involved in both and that therapeutic advances in animal studies may have human application. PMID- 11911505 TI - Age-related changes in adrenergic neuroeffector transmission. AB - In this study we have looked at the effects of ageing on prejunctional control of noradrenergic neurotransmission in the cardiovascular system, in terms of alpha2 adrenoceptors, beta2-adrenoceptors and the noradrenaline re-uptake process. These studies show diminished prejunctional alpha2- and prejunctional beta-adrenoceptor mediated responsiveness together with diminished noradrenaline re-uptake in rat tissues. The reduced prejunctional alpha2-inhibitory control and reduced re uptake found in tissues from aged rats is more than likely to outweigh the effects of reduced beta-adrenoceptor facilitation, at least in normal conditions. Hence, assuming that such changes also occur in man, we might expect to find evidence of increased release of noradrenaline from noradrenergic nerves, and this could be reflected in plasma levels of noradrenaline. PMID- 11911506 TI - Estrogen receptor downregulators: new antihormonal therapy for advanced breast cancer. AB - BACKGROUND: Fulvestrant is the first agent in the new class of estrogen receptor (ER) downregulators to be evaluated in clinical studies. The binding of ER downregulators to ERs inhibits the activation functions of the receptors (AF-1 and AF-2). ER downregulators also disrupt the dimerization and nuclear localization of ERs. In contrast to ER downregulators, when selective estrogen receptor modulators (SERMs) bind to ERs, only AF-2 is inhibited; AF-1 remains active. Therefore, with SERMs, AF-1 can still recruit coactivators, which results in a partially inactivated transcription and a reduced rate of tumor cell division. Consistent with its mechanism of action, animal models have shown that fulvestrant demonstrates a longer suppression of tumor growth than does tamoxifen, and it has antitumor activity in tamoxifen-resistant tumors. Two Phase III trials have been conducted comparing fulvestrant (250 mg IM monthly) with anastrozole (1 mg oral tablet daily) in postmenopausal patients with advanced breast cancer progressing after prior endocrine therapy. Objective response in fulvestrant-treated patients in the 2 studies ranged from 17.5% to 20.7% compared with 15.7% to 17.5% for anastrozole-treated patients, and the median duration of response ranged from 14.3 to 19.3 months in the fulvestrant group compared with 10.5 to 14.0 months in the anastrozole group. No statistically significant differences were demonstrated for any of the predefined end points. In both studies, fulvestrant was as well tolerated as anastrozole. OBJECTIVE: This review describes the pharmacology, clinical activity, safety, dosing, and pharmacokinetic profile of fulvestrant. CONCLUSIONS: Based on evidence to date, fulvestrant will likely be an important agent in the treatment of tamoxifen resistant metastatic breast cancer. PMID- 11911507 TI - Historical perspective on hormonal therapy of advanced breast cancer. AB - BACKGROUND: Endocrine therapy is the preferred first-line therapy in patients with nonaggressive, receptor-positive, metastatic breast cancer. Endocrine therapies in these patients are as effective as chemotherapy in terms of survival and tumor response. In addition, hormonal therapies produce fewer and less severe adverse effects than does chemotherapy. Classes of endocrine therapy currently on the market include selective estrogen receptor modulators (SERMs), aromatase inhibitors, and progestins. For several decades, tamoxifen has been considered the gold standard of therapy. However, despite its proven efficacy, a high proportion of patients do not respond. Therefore, a need exists for alternative therapies or for agents to use following tamoxifen failure. OBJECTIVE: This review article highlights the various endocrine options available for patients with advanced breast cancer and discusses new agents on the horizon. CONCLUSIONS: Although more studies are needed to determine the optimal sequence of hormonal therapy and the ideal agent within each class, the availability of new and multiple options is encouraging for women with advanced breast cancer. PMID- 11911508 TI - Fulvestrant: clinical application of an estrogen receptor downregulator. PMID- 11911509 TI - Cognitive assessment and neurological rehabilitation. AB - Two recent studies evaluating cognitive assessment in patients with multiple sclerosis and stroke found no benefit from providing information derived from a full cognitive assessment, and one study also failed to find any benefit following specific 'cognitive treatments'. These studies might suggest that rehabilitation teams do not need clinical psychologists. However it is already known that isolated assessment does not affect outcome, and the nature and context of the assessments and interventions in these studies simply reinforces this. Thus, given the frequency of cognitive deficits in these patients and given the strong evidence in favour of multidisciplinary teamwork in rehabilitation, these studies could be used as strong evidence that clinical psychologists should be integral members of all neurological rehabilitation teams. PMID- 11911510 TI - Evaluation of the effectiveness of professionally guided self-care for people with multiple sclerosis living in the community: a randomized controlled trial. AB - OBJECTIVE: The aim of the study was to assess the efficacy of a patient-focused professionally guided self-care programme for the management of multiple sclerosis (MS) in the community. DESIGN: This was a single-blind randomized controlled trial. SETTING: The study was conducted with people with MS living in the community. PARTICIPANTS: Two hundred and seventy-eight people with MS were invited to take part in the study. One hundred and eighty-nine people consented to take part (68%). Of these 183 began the study and 169 (92.3%) completed it. Seventy-three individuals were in the intervention group and 96 were in the control group. INTERVENTION: The intervention comprised discussion of self-care based on client priorities, using an information booklet about self-care. MAIN OUTCOME MEASURES: These included the Barthel Index, a measure of mobility, the SF 36, and the Standard Day Dependency Record (SDDR) which measures the need for assistance with daily activities. Assessments were conducted at baseline and again six months later. RESULTS: Changes in health status were small. However, at follow-up the intervention group had better SF-36 health scores, in mental health (p = 0.04), and vitality (p = 0.05) and considered help with daily activities to be less essential, as measured by the SDDR (p = 0.04), than the control group. Participants in the intervention group had maintained levels of independence at follow-up (p = 0.62) while the control group showed a significant decrease in independence (p= 0.001). CONCLUSION: This intervention could be a useful aid for health professionals who are supporting people with MS living in the community. PMID- 11911511 TI - The immediate effect of handling technique on range of movement in the hemiplegic shoulder. AB - PURPOSE: To assess the immediate effect of two different handling. techniques on range of flexion in the hemiplegic shoulder. METHOD: A randomized controlled design with within-subject comparison was used. Range of shoulder flexion was measured using a bubble goniometer. Range of passive movement was compared as the weak arm was lifted using an 'axilla hold' (when the gleno-humeral joint is supported and held in external rotation) and a 'distal hold' (when the arm is lifted at the forearm without shoulder support). Twenty-two people with arm weakness following stroke were recruited. They were inpatients or attending a day hospital in two NHS trusts, with no previous limitation of range or function of their arm. A paired t-test was used for analysis. RESULTS: Mean shoulder flexion for the axilla hold was 115.2 degrees (SD 38.45), and 97.7 degrees (SD 44.7) for the distal hold. This difference was significant at p < 0.001 (95% confidence interval (95% CI) 7.96, 26.88). CONCLUSIONS: Lifting the hemiplegic arm by holding the humerus under the axilla and maintaining external rotation produces greater range of flexion at the hemiplegic shoulder than a 'distal hold'. PMID- 11911512 TI - Effects of one-point and four-point canes on balance and weight distribution in patients with hemiparesis. AB - OBJECTIVE: To examine the effects of one-point and four-point canes on postural sway and on the distribution of weight between the lower extremities and the walking aids in hemiparetic patients. SETTING: Flieman Geriatric Rehabilitation Hospital, Haifa, Israel. SUBJECTS: Thirty hemiparetic patients following a unilateral stroke, with moderate functional impairment, and 20 age-matched healthy subjects. INTERVENTION: Subjects were tested on two forceplates, which were placed at a 30 degrees angle from each other with the heel end of the plates separated by 3 cm. Each subject was tested under three conditions: with no cane, with a one-point cane, and with a four-point cane. Testing time was 30 seconds, and order of testing was randomized. OUTCOME MEASURES: Weight borne by the lower extremities and by the walking aids expressed as a percentage of overall body weight, and Sway Index indicating vertical pressure fluctuations over both feet. RESULTS: In both subject groups, the one-point cane did not reduce sway significantly in comparison with no cane, while the four-point cane reduced sway significantly in comparison with both no cane and one-point cane. Neither cane type affected weight-bearing on the paretic leg, while significantly reducing weight-bearing on the uninvolved extremity. Mean percentage of body weight on the four-point cane was significantly higher than on the one-point cane CONCLUSIONS: A four-point cane increases stability of moderately involved hemiparetic patients during stance more than a one-point cane. The noted shift of weight toward the walking aid does not adversely affect weight-bearing on the paretic limb. PMID- 11911513 TI - Prediction of long-term functional outcome after stroke rehabilitation. AB - OBJECTIVE: To find patient characteristics in the early post stroke phase that could predict three years functional outcome. DESIGN: Prospective study. SETTING: In-hospital rehabilitation department (admission and discharge). Outpatient department one and three years post stroke. SUBJECTS: One hundred and forty-two stroke patients (56% women), median age 75 years. MAIN OUTCOME MEASURES: Barthel Index (BI) score; BI score change; accommodation status; Rankin scale score; and Frenchay Activities Index (FAI) score, all registered three years post stroke. RESULTS: The percentages of patients still living at home after one and three years were 88% and 83%, respectively. Twenty per cent of the patients had deteriorated according to the BI after three years, mostly due to recurrent strokes (odds ratio (OR) 10.3; 95% confidence interval (CI) 3.0-35.5) and co morbidity with other disabling disorders (OR 3.9; CI 1.1-13.5). Co-morbidity also emerged as an important risk factor for dependency according to BI score (OR 8.8; Cl 2.4-32.1) as well as for a poor FAI score (OR 4.9; CI 1.9-13.0). BI in the early phase was the strongest predictor for long-term functional outcome. Urinary incontinence emerged as a risk factor for nursing home placement after three years (OR 3.2; CI 0.9-11.3). Cognitive dysfunction was a risk factor for poor FAI scoring (OR 2.7; CI 1.0-7.0). CONCLUSIONS: After stroke rehabilitation, concomitant chronic disabling disorders and recurrent strokes seem to play an important role regarding dependency, handicap and long-term functional decline. PMID- 11911514 TI - Radiation-induced brachial plexopathy in women treated for carcinoma of the breast. AB - OBJECTIVES: To study the clinical presentation and natural history of radiation induced brachial plexopathy in 33 women treated for carcinoma of the breast. METHODS: All of the patients were referred to a single consultant neurologist. Details of surgical procedures, radiotherapy, symptoms at presentation and follow up and neurological findings were recorded. Patients were reviewed at six or 12 monthly intervals for 2-19 years (median 9.5 years). Investigations included blood tests, chest X-ray, bone scan, neurophysiological studies, computerized tomography (CT) or magnetic resonance imaging (MRI) of the cervical spine and cerebrospinal fluid examination. RESULTS: Symptoms began from six months to 20 years after radiotherapy (median time 1.5 years). Progressive weakness was universal and resulted in loss of any useful hand function in all but three patients. The time taken to loss of useful hand function ranged from six weeks to five years (median 1.25 years). Three patterns of upper limb weakness were identified, distal limb weakness only (13 patients), global limb weakness that was more marked distally (11 patients), and completely flaccid arm (10 patients). Seventeen patients required long-term morphine to palliate pain. A chemical sympathectomy benefited three patients. CONCLUSIONS: Most patients developed symptoms within three years, but late presentations 8-20 years later were encountered. Symptoms were progressive in all patients, though the rate did vary. Pain was common and persisted indefinitely in all but one patient. Morphine was effective and should be used early and in adequate doses. Chemical sympathectomy provided sustained relief in three of six patients. PMID- 11911515 TI - Validity of the TEMPA for the measurement of upper limb function in multiple sclerosis. AB - OBJECTIVE: To investigate concurrent and construct validity of the TEMPA (Test d'Evaluation de la performance des Membres Superieurs des Personnes Agees) in patients with upper limb dysfunction due to multiple sclerosis. SUBJECTS: Forty three patients with upper limb dysfunction due to multiple sclerosis. DESIGN: Patients performed upper limb tests and were assessed on measures of functional independence. SETTING: National Multiple Sclerosis Centre, Melsbroek, Belgium. MAIN OUTCOME MEASURES: TEMPA, Jebsen Handfunction Test, Nine Hole Peg Test, Functional Independence Measure (FIM) and activities of daily living (ADL) self questionnaire. RESULTS: The correlation between TEMPA and Jebsen Handfunction and the Nine Hole Peg test respectively is good (0.56-0.87) and high (0.79-0.9). The correlation between TEMPA and the FIM and ADL self-questionnaire was moderate (0.44-0.61) although mostly higher than the correlation between the Jebsen Handfunction and the Nine Hole Peg test respectively and the measures of functionality (0.22-0.55/0.1-0.47). CONCLUSIONS: This study provides data supporting the concurrent validity of the TEMPA with the Jebsen Handfunction and the Nine Hole Peg Test in patients with multiple sclerosis. The results suggest construct validity of the TEMPA for the measurement of functionality during activities of daily life in patients with upper limb dysfunction due to multiple sclerosis. PMID- 11911516 TI - Pitfalls in effectiveness research: a comparative analysis of treatment goals and outcome measures in stroke rehabilitation. AB - PROBLEM: There is a great diversity of movement therapies in stroke rehabilitation, each of which is treated as a more or less independent system. OBJECTIVE: To articulate pitfalls in effectiveness research that has been employed to reduce the number of different forms of treatment. METHODS: The contents of treatment goals and outcome measures in clinical and scientific texts on stroke rehabilitation were analysed and contrasted in order to uncover discrepancies. ISSUES: The main issue is that theory, and conceptualization of therapy play a diminished role in treatment effectiveness research, which may hinder the interpretation of data. The notion that the methodological and statistical tools, if correctly applied, provide researchers with the distance that is thought to be necessary for an objective judgement will be challenged. CONCLUSION: The analyses indicate that although scientifically credible measurement tools may be neutral with regard to the user, they are not necessarily neutral with regard to the therapies being compared in effectiveness research. PMID- 11911517 TI - Efficacy of two forms of electrical stimulation in increasing quadriceps strength: a randomized controlled trial. AB - OBJECTIVE: To investigate whether electrical stimulation is effective in improving quadriceps strength in healthy subjects and to compare interferential and low-frequency current in terms of the effects on quadriceps strength and perceived discomfort. DESIGN: Randomized, controlled study. SETTING: Physical Medicine and Rehabilitation Department in a university hospital. SUBJECTS: Thirty medical faculty students, divided into three groups, participated in the study. INTERVENTIONS: Group A received electrical stimulation with bipolar interferential current while group B received electrical stimulation with low frequency current (symmetrical biphasic). Group C served as the control group. Electrical stimulation was given for 15 minutes, five days a week for three weeks, at a maximally tolerated intensity with the knee fully extended in the sitting position. MAIN OUTCOME MEASURES: Before and after the study, quadriceps strength was measured with a Cybex dynamometer isokinetically at the angular velocities of 60 degrees/s and 120 degrees/s. The perceived discomfort experienced with each type of electrical stimulation was quantified by the use of a visual analogue scale (VAS). RESULTS: Statistically significant increase in isokinetic strength was observed after training in group A and group B. Increase in strength did not differ between the stimulation groups. No significant change in strength occurred in group C. Perceived discomfort by the stimulation groups was not significantly different. CONCLUSION: Both interferential and low frequency currents can be used in strength training with the parameters used in this study. PMID- 11911519 TI - Beyond the 10-m time: a pilot study of timed walks in lower limb amputees. AB - OBJECTIVE: To investigate in a pilot study the use of extended walking times as an objective method of distinguishing differences in outcome in lower limb amputees. SETTING: Sixteen lower limb amputees attending the prosthetic clinic. METHOD: Patients were asked to walk 100 m, turning every 20 m, using their usual walking aids and prostheses at their chosen walking speed. Demographic details and modified Stanmore/Harold Wood mobility grades, walking aids used and discomfort were recorded. Time to walk each 20 m up to 100 m was recorded. Statistical analysis was carried out using the Spearman rank order correlation coefficient. RESULTS: There was a wide age range and differing causes of amputation. Five out of 16 subjects failed to complete 100 m and these had the slowest 20- and 40-m times. The mean time to walk 40 m was 41 seconds for those who were able to complete 100 m and 144 seconds for those who could not complete 100 m. All those with modified Stanmore/Harold Wood mobility grades less than four failed to complete 100 m. There was a high correlation between 20-, 40-, 60 , 80- and 100-m times and mobility grades, which was statistically significant, and between 20-, 40-, 60-, 80- and 100-m times. CONCLUSION: Forty-metre walking time can differentiate between those of mobility grades 4b-6 and those with lower mobility grades who are unable to walk significantly outside. PMID- 11911518 TI - A randomized controlled trial of exercise to improve mobility and function after elective knee arthroplasty. Feasibility, results and methodological difficulties. AB - OBJECTIVE: To assess the feasibility of comparing two types of exercise regime aiming to improve mobility and function following knee arthroplasty. DESIGN: A single-blind randomized controlled trial. SUBJECTS: Patients with primary, unilateral knee osteoarthritis undergoing elective knee joint replacement. INTERVENTION: Home-based traditional exercise group (TEG) or home-based functional exercise group (FEG) following discharge from hospital. OUTCOME MEASURES: These included goniometry; a knee-specific pain score, leg extensor power and a walking test. Patients were followed up at three, six and 12 months after surgery. RESULTS: Forty-seven patients met the study criteria, 24 were randomized to the TEG and 23 to the FEG. There were marked improvements in mobility, leg extensor power and pain in the year after surgery (MANOVA p < 0.001). There were no statistically significant differences between the two exercise groups. Knee flexion decreased during the follow-up period and had not recovered by 12 months. Retention of patients was a problem, with nearly 50% lost to follow-up at 12 months. These patients were assessed as having low motivation during inpatient rehabilitation (p < 0.05). CONCLUSIONS: There were trends in favour of the FEG that were of clinical relevance. A definitive study would need a sample size of at least 100 patients in each arm. It is essential to develop strategies to combat loss to follow-up. PMID- 11911520 TI - Percutaneous endoscopic gastrostomy feeding in a district rehabilitation service. AB - OBJECTIVES: To examine gender, diagnosis, age, reasons for feeding, nutritional status, complications, outcome and duration of feeding in patients who have required a percutaneous endoscopic gastrostomy (PEG) for nutritional support at a district rehabilitation unit in the six years since the service was established. To identify potential for improvements in the management of future patients. DESIGN: Retrospective case note review of cases from 1992 to 1998. SETTING: District rehabilitation service for ages 16-64 serving the population of Fife, Scotland (population circa 350 000). SUBJECTS: All patients (n = 42) who had been fed via a PEG feeding tube in the previous six years. RESULTS: Forty-four PEG tube insertions had been conducted for 43 episodes of feeding in 42 patients. Five episodes of feeding were because of persistent vegetative state or low awareness state and 38 because of neurological swallowing impairment. Twenty-six (60%) patients were nutritionally depleted when PEG feeding was commenced. Twenty seven (64%) patients experienced minor complications and 15 (34%) had no complications. At three months post procedure four (9.5%) patients had died and 21 (50%) had been discharged home. The mean duration of feeding on 31 October 1998 of the 20 patients (48%) who continued feeding at that date was 3.19+/-1.89 (mean +/- SD) years. CONCLUSIONS: Patients requiring PEG feeding in a district rehabilitation service have a range of diagnoses and the main indication for intervention is neurological swallowing impairment. The majority of patients were nutritionally depleted when feeding commenced and the reasons for this require further investigation. PMID- 11911521 TI - The impact of contact time on pyrene sorptive behavior by a sandy-loam soil. AB - Batch experiments with pyrene (PYR) were conducted to quantify the effect of contact time on its sorption and desorption behavior by a sandy-loam soil. Twenty four and 48 h contact times were chosen for the nonequilibrium conditions and 240 h for the pseudoequilibrium study. All times was selected based on the kinetic results. The nonlinear, pseudoequilibrium sorption isotherm was fit to a two stage Freundlich model: 3-7 mg/l for the first stage and 7-15 mg/l for the second stage. A substantial fraction of the sorbed PYR was not desorbed within the given desorption time. The reason of hysteresis was found to be a sorption enhancement due to soil hydration which provided more sorption sites. A desorption enhancement at 240-h desorption steps was attributed to the increased dissolved organic matter evolution. This study also found that both soil organic matter and clay materials had an equal role in PYR sorption enhancement and desorption resistance. PMID- 11911522 TI - Recovery from acidification in Swedish forest streams. AB - Sulphur deposition in Sweden has decreased to less than half of the levels recorded in 1970 and now signs of recovery from acidification of surface waters are beginning to appear. We investigated time trends of water chemistry between 1985 and 1998 in 13 streams draining small forested catchments with generally shallow acid sensitive soils. At nine of the catchments, bulk deposition was monitored as well. Sulphate concentrations decreased in both stream water and deposition, although with somewhat smaller trends in stream water compared with deposition. The magnitude of the trends in sulphate increased from north to south, following a gradient of increasing industrial influence. Five sites in the southern half of the country showed weak signs of recovery from acidification in terms of increasing concentrations of acid neutralising capacity and decreasing concentrations of hydrogen ions, corresponding to annual increases of 0.01 pH units. Changes in stream discharge and concentrations of marine salts and organic acids could not explain the observed decrease in acidity and the results were interpreted as recovery from anthropogenic acidification. For the northern half of Sweden, any changes in water chemistry could be attributed to natural variation in climate and marine influence, and the effect of anthropogenic acidification was negligible. PMID- 11911523 TI - Organochlorine and organotin compounds in Caspian seals (Phoca caspica) collected during an unusual mortality event in the Caspian Sea in 2000. AB - Polychlorinated biphenyls (PCBs), organochlorine pesticides and organotin compounds were determined in the blubber and liver of Caspian seals (Phoca caspica) found stranded on the coast of the Caspian Sea during an outbreak of canine distemper virus (CDV) in 2000. Among organochlorines analyzed, DDTs were the most dominant contaminants with concentrations ranging from 6.3 to 470 microg/g on a lipid-weight basis. Caspian seals collected in 2000 during the epizootic had higher concentrations of organochlorines than healthy individuals sampled in 1998. However, the blubber layer was generally thinner in the seals collected in 2000 than those in the previous surveys. Although compositions of organochlorine pesticides in seals suggested that the contamination status in the Caspian Sea is improving, the levels found in Caspian seals in 2000 were comparable to those in other marine mammals that have suffered from epizootics. This implies that the present status of contamination found in Caspian seals poses a risk of immunosuppression. Concentrations of butyltin compounds in livers of seals ranged from 0.49 to 17 ng/g on a wet-weight basis and octyltin compounds were below limit of detection in all the samples analyzed, suggesting less contamination by organotin compounds in the Caspian Sea. PMID- 11911524 TI - Rice uptake and distributions of radioactive 137Cs, stable 133Cs and K from soil. AB - The distributions of radionuclides in plant components as to radionuclide transfer to animals are important for understanding the dynamics of radionuclides in an agricultural field. Most of the non-edible parts in these components are returned to the soil as organic fertilizer where they may again be utilized in the soil-plant pathway and/or are mixed with feed for livestock. Rice plants were grown in an experimental field and separated at harvest into different components, including polished rice, rice bran, hull, leaves, stem and root, and then the distributions of radioactive 137Cs, stable 133Cs and K in these components were determined. The distribution of 137Cs in polished rice and rice bran was similar to that of 133Cs, while that of K was different. The concentration ratios of 133Cs/K in leaf blade positions increased with aging, which means that the translocation rate of 133Cs in rice plants was slower than that of K. At harvest the distribution of dry weight in polished rice to entire rice plants was 34%, and the distributions of 133Cs in the polished rice and the non-edible parts were 7 and 93%, respectively, whereas those of K in the polished rice and the non-edible parts were 2 and 98%, respectively. Findings suggest that the transfer and distribution of 133Cs, not of K, provide better information on the long-term fate of 137Cs in an agricultural environment. PMID- 11911525 TI - Algal growth inhibition by river water pollutants in the agricultural area around Lake Biwa, Japan. AB - An ecotoxicological study of river water discharged from the agricultural area around Lake Biwa was performed by using algal bioassays to guide chemical analysis. Water samples were collected once a week, at least, for 1 year starting in April 1997 and continuing until April 1998. The toxicities of the dissolved and particulate-adsorbed extracts of water samples were evaluated by the algal growth inhibition test and concentrations of individual pesticides were determined. Most of the river water that was collected during the periods when pesticides were applied to the paddy fields caused algal growth inhibition. Some extracts were found to contain herbicides (molinate, mefenacet, simetryn, or esprocarb) as major compounds. According to chemical assay and bioassay, simetryn was identified as the most toxic compound that caused algal growth inhibition. PMID- 11911526 TI - Imposex and tributyltin contamination as a consequence of the establishment of a marina, and increasing yachting activities at Phuket Island, Thailand. AB - The objective was to investigate how tributyltin (TBT) contamination resulting from the establishment of a new marina and increasing yachting activities at Phuket Island, Thailand affects the area. A minimum of 30 specimen of the mangrove dwelling muricid Chicoreus capucinus were collected at five stations 2 months before and 5, 9, 18 and 33 months after Yacht Haven Marina was constructed. Imposex measured as RPLI and VDSI were significantly correlated to duration of time after construction and distance from the marina. All females developed imposex close to the marina after 18 months, whereas no significant increase occurred at stations more than 2 km from the marina. To evaluate the range of effects of the increasing yachting activities the muricid Thais distinguenda was used as an indicator of imposex at two popular mooring sites for yachts at Phi Phi and Raja Island in Phangna Bay, Thailand. The incidence of imposex was 100% at stations close to the mooring sites. A significant correlation existed between the distance from the sites and the incidence of imposex. The increasing incidence of imposex suggest that TBT contamination is worsening, against global trends, because regulations prohibiting the use of TBT based paints, do not exist in Thailand. PMID- 11911527 TI - Hazard prioritization in ecological risk assessment through spatial analysis of toxicant gradients. AB - The analysis of spatial relationships among the distribution of environmental stressors and observed or predicted adverse effects may be a useful method of prioritizing hazards in regional ecological risk assessment (ERA). Geographic information systems were used to compare the spatial distribution of toxicant concentrations in sediments of Chesapeake Bay with the distribution of areas in the basin where ecological impacts have historically been observed. Toxicants were then prioritized based upon the strength of their spatial association with the high impact areas. This method of hazard identification/prioritization was validated against the Chesapeake Bay Program's lists of toxics of concern and toxics of potential concern (TOC and TOPC, respectively). Of the 18 toxicants on the TOC/TOPC lists that were considered in the current study, 15 (83%) were identified as priority contaminants in the current study, 11 (73%) of which were either of primary or secondary concern. The use of spatial analysis tools in ERA may lead to more rapid and rigorous methods for prioritizing environmental risks. PMID- 11911528 TI - Persistent chlorinated pesticides and polychlorinated biphenyls in selected fish species from Lake Tanganyika, Burundi, Africa. AB - Concentrations of selected organochlorine pesticides and PCBs in seven fish species (cichlids) from the north end of the Lake Tanganyika, Burundi, Africa were determined. Results were compared to previous work on the Lake Tanganyika and other water bodies and to the European Community maximum residue levels (MRLs) in edible fat. The analytical method included a hot Soxhlet extraction with a mixture of acetone: hexane (1:3, v/v), gravimetrically lipid determination, and a single step clean-up. For PCBs and stable pesticides, the clean-up was done on activated silica gel impregnated with concentrated sulfuric acid, while for non acid-stable pesticides superposed layers of alumina, silica and florisil impregnated with 15% methanolic solution of KOH were successively used. Recoveries of organochlorine pesticides from certified reference material (CRM 430) were ranging from 86% for p,p'-DDT to 107% for endrin, while recoveries from blank fat spiked fortified at three different levels were between 65% for alachlor at the lowest fortification level and 107% for mirex at the highest fortification level. The limits of detection for each analyte were ranging from 0.1 ng/g to 0.5 ng/g fat. All chlorinated pesticides were found in the analyzed species but at low concentrations. Boulengerochromis microlepis contained the highest concentrations of HCHs (288.2 +/- 15.5 ng/g fat) and DDTs (909.1 +/- 42.5 ng/g fat), while the highest PCB levels (166.7 +/- 37.4 ng/g fat for the sum of 12 congeners) were found in Oreochromis niloticus. However, there is no evidence that Lake Tanganyika is more contaminated with pesticides than other African water bodies. PMID- 11911529 TI - Chlorinated and polycyclic aromatic hydrocarbons in riverine and estuarine sediments from Pearl River Delta, China. AB - Spatial distribution of chlorinated hydrocarbons [chlorinated pesticides (CPs) and polychlorinated biphenyls (PCBs)] and polycyclic aromatic hydrocarbons (PAHs) was measured in riverine and estuarine sediment samples from Pearl River Delta, China, collected in 1997. Concentrations of CPs of the riverine sediment samples range from 12 to 158 ng/g, dry weight, while those of PCBs range from 11 to 486 ng/g. The CPs concentrations of the estuarine sediment samples are in the range 6 1658 ng/g, while concentrations of PCBs are in the range 10-339 ng/g. Total PAH concentration ranges from 1168 to 21,329 ng/g in the riverine sediment samples, whereas the PAH concentration ranges from 323 to 14,812 ng/g in the sediment samples of the Estuary. Sediment samples of the Zhujiang River and Macao harbor around the Estuary show the highest concentrations of CPs, PCBs, and PAHs. Possible factors affecting the distribution patterns are also discussed based on the usage history of the chemicals, hydrologic condition, and land erosion due to urbanization processes. The composition of PAHs is investigated and used to assess petrogenic, combustion and naturally derived PAHs of the sediment samples of the Pearl River Delta. In addition, the concentrations of a number of organic compounds of the Pearl River Delta samples indicate that sediments of the Zhujiang river and Macao harbor are most likely to pose biological impairment. PMID- 11911530 TI - Butyltin residues in blue mussels (Mytilus edulis) and arkshells (Scapharca broughtonii) collected from Korean coastal waters. AB - Butyltin compounds (BTs) including tributyltin (TBT) and its degradation products, di- (DBT) and mono-butyltin (MBT), were determined in bivalves such as blue mussels (Mytilus edulis) and arkshells (Scapharca broughtonii) collected from Korean coastal waters. BTs were detected in all the blue mussels and arkshells analyzed. The concentrations of total butyltin (sigmaBTs: MBT + DBT + TBT) in blue mussels and arkshells ranged from 49 to 2500 ng/g and 29 to 87 ng/g wet weight, respectively. Higher concentrations of BTs were found in blue mussels collected from Okpo and Kohyonsong Bays and Jangsengpo Harbor where large shipyards and harbors are located with dry-dock facilities. This suggested that maritime activities nearby the harbors play a major role as the source of BTs. Concentrations of TBT in mussels collected from Korea were one of the highest values reported, suggesting ongoing TBT contamination in Korea. Among BTs, TBT was the predominant compound both in blue mussels and arkshells collected from almost all the sampling locations, indicating the fresh input of TBT in Korean coastal waters. Smaller mussels tended to accumulate BTs at higher concentrations than larger ones, which may be due to higher filtration rate of small mussels and/or contact with surface microlayer in intertidal zones. PMID- 11911531 TI - Is metal extraction by Arabidopsis halleri related to exchangeable metal rates in soils amended with different metal-bearing solids? AB - Metals are associated to various constituents in polluted soils, and their availability is closely related to their chemical speciation. Studies on relations between metal extraction efficiency by hyperaccumulators and location of metals with respect to soil constituents are scarce. In this study. we investigate the relationship between metal extraction by Arabidopsis halleri and the exchangeable metals from substrates amended with various metal-bearing solids collected in the vicinity of a Zn smelter complex. These consisted of fresh and decomposing organic matter, the soil clay fraction, and two types of waste slags. ZnSO4 was also used as metal-bearing solid. Each was mixed with an unpolluted soil to produce two types of substrate, one moderately polluted and the other highly polluted. Total Zinc, Cd, Cu, and Pb were measured in substrates and in roots and shoots of A. halleri. Analysis of 0.01 M CaCl2 exchangeable metals in each substrate was performed before and after plant growth. The results showed different concentrations of exchangeable metals after plant growth, depending on the nature of the metal-bearing solids. In the ZnSO4 soil substrate, the proportion of exchangeable Zn decreased after plant growth, whilst it increased significantly on substrates amended with the two waste slags. For the other substrates, exchangeable Zn was not significantly different before and after plant growth. The same trend was observed for Cd. In the case of Cu, exchangeable rates increased in all substrates. The results were discussed according to the characteristics of the metal-bearing solids and to the metal-uptake strategy of A. halleri. PMID- 11911532 TI - High contents of rare earth elements (REEs) in stream waters of a Cu-Pb-Zn mining area. AB - Stream waters draining an old mining area present very high rare earth element (REE) contents, reaching 928 microg/l as the maximum total value (sigmaREE). The middle rare earth elements (MREEs) are usually enriched with respect to both the light (LREEs) and heavy (HREEs) elements of this group, producing a characteristic "roof-shaped" pattern of the shale Post-Archean Australian Shales normalized concentrations. At the Fenice Capanne Mine (FCM), the most important base metal mine of the study area, the REE source coincides with the mine tailings, mostly the oldest ones composed of iron-rich materials. The geochemical history of the REEs released into Noni stream from wastes in the FCM area is strictly determined by the pH, which controls the REE speciation and in-stream processes. The formation of Al-rich and mainly Fe-rich flocs effectively scavenges the REEs, which are readily and drastically removed from the solution when the pH approaches neutrality. Leaching experiments performed on flocs and waste materials demonstrate that Fe-oxides/oxyhydroxides play a key role in the release of lanthanide elements into stream waters. The origin of the "roof shaped" REE distribution pattern as well as the peculiar geochemical behavior of some lanthanide elements in the aqueous system are discussed. PMID- 11911533 TI - The role of dissolved organic carbon in the mobility of Cd, Ni and Zn in sewage sludge-amended soils. AB - A pot experiment was conducted to investigate the effect of application of naturally derived dissolved organic compounds (DOC) on the uptake of Cd, Ni and Zn by Lolium perenne L. from mixtures of soil and sewage sludge and on their extractability with CaCl2. DOC was applied at concentrations of 0, 285 and 470 mg l(-1) to a loamy sand (LS) and a sandy clay loam (SCL) soil mixed with sewage sludge at rates equivalent to 0, 10 and 50 t ha(-1). DOC applications significantly increased the extractability of metals and also their uptake by ryegrass, but the increase was greater where sludge was applied at 50 t ha(-1). It is suggested that DOC in soils significantly increased the availability of the metals to plants. This was especially the case in the LS soil, where DOC had less competition with surface sorption than in the SCL soil. PMID- 11911534 TI - What is cosmetic surgery? PMID- 11911535 TI - Chemical peels. PMID- 11911536 TI - Regional anesthesia for aesthetic surgery. PMID- 11911537 TI - Scar management: keloid, hypertrophic, atrophic, and acne scars. PMID- 11911538 TI - Leg vein management: sclerotherapy, ambulatory phlebectomy, and laser surgery. PMID- 11911539 TI - Anterior transposition of the inferior oblique muscle for treatment of superior oblique palsy. AB - PURPOSE: Weakening of the inferior oblique muscle is the procedure of primary importance in patients with superior oblique palsy, Knapp's Classes I and III. In this study, the effectiveness of anterior transposition of the inferior oblique muscle in treatment of these patients was evaluated. METHODS: Sixteen patients with superior oblique palsy, Knapp's Classes I and III, underwent anterior transposition of the inferior oblique muscle. The tip of the disinserted muscle was sutured to the sclera, parallel, and adjacent to the lateral border of the inferior rectus muscle insertion. The prism and alternate cover test measurements were made in all cardinal positions of gaze before and 6 months after surgery. RESULTS: The mean reduction of hyperdeviation was 15 prism diopters (PD) in the primary position, 23.4 PD in adduction, 26.65 PD in elevation and adduction, and 18.63 PD in depression and adduction. There was no hypotropia in the primary position. Mild limitation of upgaze has occurred in 3 of these patients, and mild fullness of the lower lid was developed by 25%. Postoperative hyperdeviation in the primary position was 5 PD or less in 15 out of 16 patients. CONCLUSIONS: The anterior transposition of the inferior oblique muscle is very effective in eliminating hyperdeviation in patients with superior oblique palsy, Knapp's Classes I and III. Up to 25 PD reduction of hyperdeviation in the primary position can be achieved. If this type of anterior transposition is used, primary position hypotropia or marked limitation of upgaze possibly will not occur. PMID- 11911540 TI - Lack of response to chemoreduction in presumed well differentiated retinoblastoma. PMID- 11911542 TI - Bilateral multiple orbital Lymphangiomas in a patient with systemic lymphangiomatosis. PMID- 11911541 TI - Cat-inflicted corneal laceration: a presentation of two cases and a discussion of infection-related management. PMID- 11911543 TI - Increasing hyperopia and esotropia as the presenting signs of bilateral diffuse choroidal hemangiomas in a patient with Sturge-Weber syndrome. PMID- 11911544 TI - Central retinal artery occlusion in herpes zoster ophthalmicus. PMID- 11911545 TI - Xerophthalmia secondary to short bowel syndrome. PMID- 11911546 TI - The sensitive period of visual development in humans. PMID- 11911547 TI - Long-term rates of PCO following small incision foldable acrylic intraocular lens implantation in children. AB - PURPOSE: To evaluate the long-term incidence of postoperative posterior capsular opacification (PCO) in children undergoing small incision foldable acrylic lens implantation with at least 2 years of follow up. METHODS: In 18 children, 26 eyes underwent small incision cataract extraction with posterior chamber foldable acrylic lens implantation. The posterior capsule was left intact in all patients at the time of surgery. RESULTS: With a mean follow up of 2.75 years and a mean age at surgery of 8.25 years, 13 of 26 eyes (50%) developed visually significant PCO requiring intervention. In the group of children under 4 years of age, 5 of 5 eyes (100%) developed visually significant PCO, while 8 of 21 eyes (38%) in the group of children over 4 years of age developed opacification. Four of 26 eyes (15%) required two procedures (either repeat Nd:YAG laser capsulotomy or pars plana secondary membrane removal) to clear the visual axis. CONCLUSION: In this study, the incidence of PCO following small incision acrylic lens implantation in children over 4 years of age is lower than those rates reported by conventional large incision rigid lens techniques with a minimum of 2 years follow up. This technique has advantages over conventional techniques in older children because it offers less surgical intervention, a lower cost to patients, and less risk of vitreous and retinal complications. PMID- 11911548 TI - Extraocular muscle surgery in dysthyroid orbitomyopathy: influence of previous conditions on surgical results. AB - PURPOSE: To determine the influence of previous treatments other than the surgical treatment of restrictive myopathy resulting from dysthyroid orbitomyopathy. METHODS: The outcome of 23 cases with dysthyroid orbitomyopathy was evaluated after extraocular muscle surgery. RESULTS: It has been observed that the duration of the orbitomyopathy, severity of myopathy of the deviation angle, and the modality of treatment prior to surgery (radiotherapy and corticotherapy) did not influence surgical outcome. Binocular single vision was achieved in primary and reading positions in 14 of 23 cases (60.9%) after surgery while prisms were required to achieve the same result in 5 cases (21.7%). CONCLUSION: It is concluded that previous treatment does not influence the results of extraocular muscle surgery in patients with dysthyroid orbitomyopathy. PMID- 11911549 TI - Effect of levodopa and carbidopa in human amblyopia. AB - BACKGROUND: To assess the role of continuous therapy for 3 weeks with levodopa and carbidopa in the management of human amblyopia in children and adults. METHOD: There were 88 amblyopic eyes of 82 subjects included in this double masked randomized prospective clinical trial. Levodopa and carbidopa combination in 2 different dosage schedules were given to both adults and children. The response was monitored of the improvement in visual acuity, contrast sensitivity, and visually evoked potentials. RESULT: Patients receiving higher dosages of levodopa and carbidopa in both adults and children showed a better response to treatment. However, the effect did not last beyond 9 weeks of stopping treatment. CONCLUSION: Though levodopa and carbidopa therapy may not be able to ameliorate amblyopia on its own on a long-term basis, it may be considered nonetheless to be an important adjunct to conventional therapy because it may improve patient compliance for occlusion by improving visual acuity in the amblyopic eye. Thus, it offers promise of improving the functional outcome in these cases. However, longer follow-up trials are needed to substantiate these conclusions. PMID- 11911550 TI - Long-term results of superior oblique tendon elongation for Brown's syndrome. AB - PURPOSE: To evaluate superior oblique tendon lengthening technique in the treatment of congenital Brown's Syndrome. METHODS: Retrospective evaluation of 8 patients with Brown's syndrome (ages 2.5 to 8 years) who underwent superior oblique split tendon elongation as the primary procedure. RESULTS: The mean follow-up period was 35 months (range 18 to 72 months). Seven of 8 patients had complete resolution of the compensatory head posturing and downshoot in adduction, and achieved some ability to elevate the eye in adduction. One of the operated eyes required reoperation because head posture persisted and a tenectomy of the superior oblique tendon was performed with satisfactory results. CONCLUSIONS: The superior oblique split tendon lengthening technique is proposed as a possible procedure of choice for the treatment of Brown's syndrome. PMID- 11911551 TI - Vertical rectus recession for the innervational upshoot and downshoot in Duane's retraction syndrome. AB - PURPOSE: To report the results of recession of the vertical rectus muscle for the innervational upshoot and downshoot in Duane's retraction syndrome. METHODS: Ten patients who had Duane's retraction syndrome with innervational upshoot or downshoot underwent recession of the superior and inferior rectus muscle for the upshoot and downshoot, respectively. This procedure was combined with recession of the lateral rectus muscle(s) for exotropia in 6 patients and for the mechanical upshoot-downshoot in one patient. Postoperatively, the effects of surgery on the upshoot/downshoot, and horizontal and vertical deviation in the primary position were recorded. Average follow-up period was 1.2 years. RESULTS: Following surgery, the innervational upshoot/downshoot was eliminated in all patients. Mean vertical deviation in the primal position in 6 patients was reduced from 21.2 to 2.5 prism diopters and none of them developed a consecutive vertical imbalance. Four patients did not have a vertical deviation in the primary position preoperatively and one of them developed 10 prism diopters hypotropia following recession of the superior rectus muscle. CONCLUSIONS: Recession of the superior and inferior rectus muscle is a safe and effective treatment for the innervational upshoot and downshoot, respectively, in Duane's retraction syndrome. PMID- 11911552 TI - Dosing and switching strategies for quetiapine fumarate. AB - BACKGROUND: The atypical antipsychotic agent quetiapine fumarate has demonstrated efficacy and tolerability in clinical trials in patients with chronic or subchronic exacerbations of schizophrenic symptoms. OBJECTIVE: This review summarizes clinical trial data and other practical information regarding the initiation and routine administration of quetiapine. Appropriate strategies for switching from other antipsychotic agents to quetiapine are also discussed. RESULTS: Quetiapine is an appropriate initial treatment for psychotic disturbances in patients with schizophrenia of any stage and for those in whom a therapeutic switch is indicated for clinical reasons, such as inability to tolerate the side effects of treatment. Titration to 400 mg/d is recommended using the following schedule, administered BID in divided doses: day 1, 50 mg; day 2, 100 mg: day 3, 200 mg; day 4, 300 mg; and day 5, 400 mg. In patients who respond to quetiapine, therapy should be continued at the optimal dose that maintains remission, within the range of 150 to 750 mg/d. When a change in therapy is indicated, several strategies for switching from one antipsychotic agent to another may be applied to switching to quetiapine. Whereas studies have shown that an abrupt switch to or withdrawal from quetiapine does not produce significant clinical consequences, in practice the switch should be carefully individualized to minimize the potential for psychotic relapse or development of withdrawal symptoms. CONCLUSIONS: Quetiapine has antipsychotic effects and good tolerability at doses from 150 to 750 mg/d. Patients can be switched to quetiapine and their treatment individualized to achieve the optimal therapeutic effect with a minimum of dose-limiting side effects. There are several strategies for switching to quetiapine from another antipsychotic agent that do not appear to cause significant exacerbation of psychosis or withdrawal reactions. PMID- 11911553 TI - A randomized, double-blind, multicenter comparison of gatifloxacin versus ciprofloxacin in the treatment of complicated urinary tract infection and pyelonephritis. AB - BACKGROUND: Gatifloxacin is a fluoroquinolone antibiotic with a broad spectrum of in vitro and in vivo activity against the gram-negative and gram-positive pathogens frequently implicated in urinary tract infections (UTIs). OBJECTIVE: This study compared the clinical and bacteriologic efficacy and tolerability of gatifloxacin versus ciprofloxacin in adult patients with complicated UTIs or pyelonephritis. METHODS: In this double-blind, multicenter, randomized, comparative study, patients were treated with either gatifloxacin 400 mg once daily or ciprofloxacin 500 mg twice daily for 7 to 10 days. Bacteriologic eradication (by quantitative urine culture) and clinical efficacy rates were assessed at a test-of-cure visit (5 to 9 days and 4 to 11 days posttreatment, respectively) and at an extended follow-up visit (29-42 days and 25-50 days posttreatment, respectively). RESULTS: A total of 372 adults were randomized to treatment, 189 to gatifloxacin and 183 to ciprofloxacin. The most commonly isolated pretreatment pathogens (n = 292) were Escherichia coli (53%) and Klebsiella pneumoniae (13%). Pathogen eradication rates for complicated UTIs were 92% and 83% with gatifloxacin and ciprofloxacin, respectively (95% CI, -4.1% to 24.5%); for pyelonephritis, the respective rates were 92% and 85% (95% CI, -20% to 37%). Clinical response rates of >90% were observed in both treatment groups among patients with complicated UTIs as well as those with pyelonephritis. Sustained eradication rates were 76% (64/84) with gatifloxacin and 66% (52/79) with ciprofloxacin. Both drugs were well tolerated, with the most common adverse events in both treatment groups being nausea, dizziness, diarrhea, and vomiting. CONCLUSIONS: Gatifloxacin is comparable to ciprofloxacin based on clinical efficacy and bacteriologic eradication rates for the treatment of complicated UTIs or pyelonephri- tis and is associated with a low incidence of clinically significant adverse events. PMID- 11911555 TI - Assessment of antihypertensive drug use in primary care in Ravenna, Italy, based on data collected in the PANDORA project. AB - BACKGROUND: In the clinical-practice setting, only a small percentage of patients treated with antihypertensive drugs become normotensive. Furthermore, the diversity of drug classes used makes comparison of treatments difficult. OBJECTIVE: The goal of this study was to characterize the types and efficacy of antihypertensive treatments used in primary care in the area of Ravenna, Italy. The study was conducted in the context of the PANDORA Project, an open-ended global outcome study. METHODS: Data were gathered from general practitioners (GPs) and were stored by the GPs or through links with national health service databases. The population of interest was patients with essential hypertension taking antihypertensive medication, each of whom was observed for 365 days. Blood pressure was measured in the morning or afternoon using an automated device, in accordance with normal clinical practice. At each office visit, the GP reviewed and made any necessary adjustments to the patient's antihypertensive treatment. Antihypertensive drug use was assessed by calculating the mean daily dose (MDD) of the prescribed drug and the duration of treatment (DT). A DT > or = 273 days constituted continuous therapy, and a DT <273 days constituted discontinuous therapy. Adverse events were not collected. RESULTS: Twenty-one GPs took part in the study. The study population included 969 patients (443 men, 526 women), all of them white, whose ages ranged from 23 to 88 years. At enrollment, 327 patients were normotensive (blood pressure <140/90 mm Hg) and 642 were hypertensive despite drug treatment. More than 25 treatment regimens were identified. Over the course of follow-up, 49 patients had discontinuous therapy and 920 had continuous therapy. Among those who had continuous therapy, 117 (12.7%) took an MDD of <0.5 tablet/d; 297 (32.3%) took > or = 0.5 and <1 tablet/d; 364 (39.6%) took > or = 1 and <2 tablets/d; and 142 (15.4%) took > or = 2 tablets/d. At the end of the observation period, the proportion of normotensive patients had increased by 5.7% (P < 0.001). CONCLUSION: Based on the findings of this study, improper use of antihypertensive drug therapy appears to be one of the reasons for the relatively small proportion of patients who attain blood pressure control with treatment. PMID- 11911554 TI - Prevalence and patterns of concomitant use of selective serotonin reuptake inhibitors and other antidepressants in a high-cost polypharmacy cohort. AB - BACKGROUND: Concomitant antidepressant therapy for patients who do not respond to selective serotonin reuptake inhibitors (SSRIs) may be appropriate under close medical supervision. However, little is known about the prevalence or patterns of concurrent antidepressant therapy in a typical large health maintenance organization. OBJECTIVE: The purpose of this study was to determine the prevalence of concomitant SSRI-antidepressant therapy and to assess the relationship between concomitant SSRI therapy, patient demographic characteristics, and the use of multiple prescribers and pharmacies. METHODS: This was a retrospective analysis of administrative prescription and medical claims data from January 1998 through September 1999. Data were obtained on beneficiaries who had >15 prescriptions dispensed in either of the first 2 quarters of 1999 and/or patients who accrued >$1,000 in prescription costs in either or both of the quarters. Patients were defined as undergoing concomitant SSRI therapy if they had received > or = 14 days of concomitant treatment with 2 SSRIs, an SSRI and tricyclic antidepressant, an SSRI and benzodiazepine, or an SSRI and miscellaneous antidepressant. Contingency analysis and logistic regression were used to identify factors associated with concomitant SSRI therapy. RESULTS: The relative risk for concomitant SSRI-SSRI therapy for patients with multiple prescribers versus a single prescriber was 2.32; the relative risk for patients receiving prescriptions from multiple pharmacies versus a single pharmacy was 2.97. Female patients were 19.8% more likely than male patients to receive concomitant SSRI therapy. Use of multiple prescribers increased the odds for concomitant SSRI therapy by >3.0 across the 4 therapeutic combinations. Use of multiple pharmacies increased the odds for concomitant SSRI SSRI therapy by 5.42. CONCLUSIONS: Prescription of concomitant SSRI therapy was strongly associated with changes in strength of dosage and products and with use of multiple prescribers and pharmacies. PMID- 11911556 TI - Pharmacokinetic evaluation of a sprinkle-dose regimen of a once-daily, extended release morphine formulation. AB - BACKGROUND: Morphine sulfate extended-release (MSER) uses a drug-delivery technology that allows once-daily dosing. It is possible to open the MSER capsule and sprinkle the contents on soft food, a potentially useful alternative to the intact capsule in patients who have difficulty swallowing. OBJECTIVE: This study compared the bioavailability of MSER and its metabolites morphine-3-glucuronide and morphine-6-glucuronide after administration of MSER in a sprinkle-dose regimen relative to an intact capsule swallowed whole. METHODS: This was a randomized, open-label, single-dose, crossover study, with a 7-day washout period between the 2 dosing days. Healthy volunteers were randomized to receive an intact 60-mg MSER capsule swallowed whole or the contents of a 60-mg MSER capsule sprinkled on applesauce. Blood samples were collected and analyzed for concentrations of morphine and its active glucuronide metabolites. Pharmacokinetic (PK) parameters were calculated and bioequivalence assessed. Bioequivalence was concluded if the 90% CIs of the ratio of log-transformed values for maximum concentration (Cmax) and area under the plasma concentration time curve (AUC) were within 80% to 125%. RESULTS: Of 30 subjects enrolled, 28 completed the study and were eligible for PK evaluation. Two subjects were withdrawn for reasons unrelated to study treatment. The plasma concentration-time profiles of morphine and its metabolites were superimposable after administration of the 2 regimens. Cmax and total systemic exposure-based on AUC from time 0 to the last quantifiable concentration (AUC(last)) and AUC from time 0 to infinity (AUC(infinity))-were comparable between treatments. The 90% CIs for morphine AUC(last), AUC, and Cmax ratios were 98 to 109, 96 to 106, and 95 to 117, respectively. Similar 90% CIs were obtained for the morphine metabolites. CONCLUSION: In this study in healthy volunteers, sprinkling the entire contents of an MSER capsule onto applesauce and swallowing without chewing was bioequivalent to swallowing an intact MSER capsule. PMID- 11911557 TI - A multicenter, randomized, investigator-blinded study of 5- and 10-day gatifloxacin versus 10-day amoxicillin/clavulanate in patients with acute bacterial sinusitis. AB - BACKGROUND: Treatment guidelines for acute bacterial sinusitis recommend 10 to 14 days of therapy with amoxicillin/clavulanate, high-dose amoxicillin, cefpodoxime, cefuroxime, or a newer fluoroquinolone. OBJECTIVE: This study compared the clinical efficacy of short-course (5-day) gatifloxacin with standard 10-day regimens of amoxicillin/clavulanate or gatifloxacin in patients with a diagnosis of acute, uncomplicated maxillary sinusitis. METHODS: This was a multicenter, investigator-blinded study in adult patients (age >18 years) with physical findings, signs and symptoms (for at least 7 days), and radiographic findings indicating acute, uncomplicated maxillary sinusitis. Patients were randomized to receive gatifloxacin 400 mg once daily for 5 days, gatifloxacin 400 mg once daily for 10 days, or amoxicillin/clavulanate 875 mg twice daily for 10 days. Clinical response was assessed once between days 3 and 5 of treatment, once I to 3 days after the completion of study treatment, once 7 to 14 days posttreatment (test-of cure visit), and once 21 to 28 days posttreatment. Safety was assessed throughout the study. RESULTS: The study enrolled 445 patients. The treatment groups were similar in terms of history of sinusitis, presenting signs and symptoms, and radiographic findings. The most common presenting symptoms were nasal congestion, sinus tenderness, and purulent nasal discharge (>90% of patients); 99% of patients had abnormal radiographic findings. At the test-of-cure visit, clinical cure rates for clinically evaluable patients in the 3 treatment groups were 74% (102/137) for 5-day gatifloxacin, 80% (101/127) for 10-day gatifloxacin, and 72% (101/ 141) for 10-day amoxicillin/clavulanate (95% CI for the difference in cure rates: 5-day gatifloxacin vs amoxicillin/clavulanate, -7.6 to 13.2; 5- vs 10-day gatifloxacin, -15.2 to 5.1; 10-day gatifloxacin vs amoxicillin/clavulanate, -2.3 to 18.1). The distribution and incidence of drug-related adverse events (AEs) were comparable between treatment groups, and the majority (>95%) were mild or moderate in severity. The most common drug-related AEs included vaginitis, diarrhea, and nausea. CONCLUSION: In this population of patients with acute, uncomplicated sinusitis of presumed bacterial origin, a short course (5 days) of gatifloxacin therapy was associated with comparable clinical cure rates and tolerability to those of standard 10-day therapy with gatifloxacin or amoxicillin/clavulanate. PMID- 11911558 TI - Tramadol/acetaminophen combination tablets for the treatment of osteoarthritis flare pain: a multicenter, outpatient, randomized, double-blind, placebo controlled, parallel-group, add-on study. AB - BACKGROUND: In a flare of osteoarthritis (OA) pain, increasing the dose of standard anti-inflammatory or routine analgesic drugs may not be practical because of an increased incidence of side effects. In patients achieving inadequate pain relief from traditional non-steroidal anti-inflammatory drugs (NSAIDs) or cyclooxygenase (COX)-2-selective inhibitors, it may be appropriate to add an analgesic agent with a different mechanism of action, thereby targeting multiple components of the pain pathway. OBJECTIVE: The addition of tramadol/acetaminophen tablets to existing therapy was compared with the addition of placebo in the treatment of OA flare pain. METHODS: This was a multicenter, outpatient, randomized, double-blind, placebo-controlled, parallel-group, add-on study. Patients received 1 or 2 tramadol/acetaminophen (37.5 mg/325 mg) tablets QID or matching placebo for 10 days in addition to ongoing NSAID or COX-2 selective inhibitor therapy. The primary outcome measures were average daily pain intensity and average daily pain relief scores from days 1 through 5. RESULTS: Three hundred eight patients were randomized to tramadoUacetaminophen (n = 197) or placebo (n = 111) and were followed for up to 10 days. Patients had a mean (+/ SD) age of 60.1 +/- 9.87 years, and were predominantly female (71.8%) and white (87.7%). Their mean (+/- SD) pain visual analog score at baseline was 73.2 +/- 11.8 mm, and their mean pain intensity score was 2.4 +/- 0.5 (on a scale from 0 = none to 3 = severe). Average daily pain intensity and pain relief scores were significantly improved with tramadol/acetaminophen compared with placebo on the primary assessment of efficacy from days 1 through 5 (both, P < 0.001) and on the assessment of efficacy from days I through 10 (both, P < 0.001) Tramadol/acetaminophen was significantly superior to placebo on the patients' and physicians' overall assessments of medication (both, P < 0.001) and on 3 of 4 subscales (pain [P = 0.004], physical function [P = 0.013], and overall [P = 0.008]) of the Western Ontario and McMaster Universities Osteoarthritis Index Questionnaire. The most common treatment-emergent adverse events with tramadol/acetaminophen were nausea, vomiting, and dizziness. No serious adverse events were reported in the tramadol/acetaminophen group. CONCLUSION: In this study, addition of tramadol/acetaminophen to NSAID or COX-2-selective inhibitor therapy was well tolerated and effective in the treatment of OA flare pain. PMID- 11911559 TI - Medical spending increases, but pharmaceuticals pose a paradox. PMID- 11911560 TI - Relationship between daily dose frequency and adherence to antihypertensive pharmacotherapy: evidence from a meta-analysis. AB - BACKGROUND: Rates of patient adherence (compliance) to pharmacotherapy range from <5% to >90%. Negative determinants include multiple daily dosing (MDD), chronic duration, and asymptomatic disease. Reports suggest that once-daily (QD) dosing may improve adherence, but their findings are inconclusive. OBJECTIVE: The purpose of this study was to compare the rates of adherence with QD, twice-daily (BID), and MDD antihypertensive drug regimens. METHODS: MEDLINE, Embase, and International Pharmaceutical Abstracts databases were searched to identify comparative trials of patient adherence to antihypertensive medication in solid, oral formulations. Data were combined using a random-effects meta-analytic model. RESULTS: Eight studies involving a total of 11,485 observations were included (1,830 for QD dosing, 4405 for BID dosing, 4147 for dosing >2 times daily [>BID], and 9655 for MDD), in which the primary objective was to assess adherence. The average adherence rate for QD dosing (91.4%, SD = 2.2%) was significantly higher (Z = 4.46, P < 0.001) than for MDD (83.2%, SD = 3.5%). This rate was also significantly higher (Z = 2.22, P = 0.026) than for BID dosing (92.7% [SD = 2.3%] vs 87.1% [SD = 2.9%]). The difference in adherence rates between BID dosing (90.8%, SD = 4.7%) and >BID dosing (86.3%, SD = 6.7%) was not significant (Z = 1.82, P = 0.069). CONCLUSIONS: The results of this meta-analysis demonstrate that with antihypertensive medications, QD dosing regimens are associated with higher rates of adherence than either BID or MDD regimens. PMID- 11911561 TI - Impact of smoking on respiratory illness-related outpatient visits among 50- to 75-year-olds in the United States. AB - BACKGROUND: Although it is generally agreed that tobacco use poses an enormous public health problem, payment and reimbursement for smoking-cessation interventions by financially stretched national health systems remain controversial. OBJECTIVE: The purpose of this study was to estimate the number and cost of excess respiratory illness-related visits attributable to smoking among older adults in the United States. METHODS: The 1995 and 1996 National Ambulatory Medical Care Survey databases were analyzed to estimate attributable risk in the population by age, sex, and smoking status among adults 50 to 75 years of age. Cost estimates for ambulatory physician visits were based on data from a major New England insurer using combined 1995 and 1996 information. Cost estimates were then developed for patients who had a respiratory-illness related diagnosis. RESULTS: Smoking was responsible for 5.1% (1,358,565) and 5.7% (1,452,761) of respiratory illness-related physician visits in 1995 and 1996, respectively. The costs (in 1998 dollars) of physician visits attributable to smoking were $69,205,301 and $74,003,645 in 1995 and 1996, respectively. CONCLUSIONS: Smoking increases health services utilization related to respiratory illness, thereby substantially increasing health care costs. Smoking-cessation programs may help reduce physician office visits related to respiratory illness, as well as the overall societal burden of smoking. PMID- 11911563 TI - Concerned with protocol used in urine production study. PMID- 11911562 TI - Effectiveness of low doses of simvastatin versus atorvastatin: resolving conflicting evidence. PMID- 11911564 TI - A method of probability diagnostic assignment that applies bayes theorem for use in serologic diagnostics, using an example of Neospora caninum infection in cattle. AB - OBJECTIVE: To develop a method of probability diagnostic assignment (PDA) that uses continuous serologic measures and infection prevalence to estimate the probability of an animal being infected, using Neospora caninum as an example. ANIMALS: 196 N caninum-infected beef and dairy cattle and 553 cattle not infected with N caninum; 50 dairy cows that aborted and 50 herdmates that did not abort. PROCEDURE: Probability density functions corresponding to distributions of N caninum kinetic ELISA results from infected and uninfected cattle were estimated by maximum likelihood methods. Maximum likelihood methods also were used to estimate N caninum infection prevalence in a herd that had an excessive number of abortions. Density functions and the prevalence estimate were incorporated into Bayes formula to calculate the conditional probability that a cow with a particular ELISA value was infected with N caninum. RESULTS: Probability functions identified for infected and uninfected cattle were Weibull and inverse gamma functions, respectively. Herd prevalence was estimated, and probabilities of N caninum infection were determined for cows with various ELISA values. CONCLUSIONS AND CLINICAL RELEVANCE: Use of PDA offers an advantage to clinicians and diagnosticians over traditional seronegative or seropositive classifications used as a proxy for infection status by providing an assessment of the actual probability of infection. The PDA permits use of all diagnostic information inherent in an assay, thereby eliminating a need for estimates of sensitivity and specificity. The PDA also would have general utility in interpreting results of any diagnostic assay measured on a continuous or discrete scale. PMID- 11911565 TI - Density of corneal endothelial cells, corneal thickness, and corneal diameters in normal eyes of llamas and alpacas. AB - OBJECTIVE: To determine density of corneal endothelial cells, corneal thickness, and corneal diameters in normal eyes of llamas and alpacas. ANIMALS: 36 llamas and 20 alpacas. PROCEDURE: Both eyes were examined in each camelid. Noncontact specular microscopy was used to determine density of corneal endothelial cells. Corneal thickness was measured, using ultrasonographic pachymetry. Vertical and horizontal corneal diameters were measured, using Jameson calipers. RESULTS: Values did not differ significantly between the right and left eyes from the same camelid. There was no significant effect of sex on density of corneal endothelial cells or corneal thickness in either species. Mean density of endothelial cells was 2,669 cells/mm2 in llamas and 2,275 cells/mm2 in alpacas. Density of endothelial cells decreased with age in llamas. Polymegathism was observed frequently in both species. Mean corneal thickness was 608 microm for llamas and 595 microm for alpacas. Corneal thickness and density of endothelial cells were negatively correlated in llamas. Older (> 36 months old) llamas had significantly larger horizontal and vertical corneal diameters than younger llamas, and older alpacas had a significantly larger vertical corneal diameter than younger alpacas. CONCLUSIONS AND CLINICAL RELEVANCE: Density of corneal endothelial cells is only slightly lower in camelids than other domestic species. Density of endothelial cells decreases with age in llamas. Age or sex does not significantly affect corneal thickness in normal eyes of llamas and alpacas. Specular microscopy is useful for determining density of corneal endothelial cells in normal eyes of camelids. PMID- 11911566 TI - Associations between dry dietary factors and canine calcium oxalate uroliths. AB - OBJECTIVE: To identify factors in dry diets associated with the occurrence of calcium oxalate (CaOx) uroliths in dogs. ANIMALS: 600 dogs with CaOx uroliths and 898 dogs without urinary tract diseases. PROCEDURE: Univariate and multivariate logistic regression were performed. RESULTS: Compared with diets with the highest concentrations of sodium, dry diets with the lowest concentrations of sodium, phosphorus, calcium, chloride, protein, magnesium, or potassium were linearly associated with increased risk of CaOx urolith formation. Significant nonlinear associations between increased occurrence of CaOx uroliths and urine acidifying potential and low moisture content were observed. Significant nonlinear associations between decreased occurrence of CaOx uroliths and carbohydrate and fiber contents were observed. A significant association between the occurrence of CaOx uroliths and dietary fat was not observed. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dry diets formulated to contain high concentrations of protein, calcium, phosphorus, magnesium, sodium, potassium, and chloride may minimize formation of CaOx uroliths. In addition, comparison of risk and protective factors of various diet ingredients fed to dogs with CaOx uroliths suggests that although similar findings were observed in canned and dry formulations, in general, greater risk is associated with dry formulations. However, before these hypotheses about dietary modifications are adopted by food manufacturers, they must be investigated by use of appropriately designed clinical studies of dogs with CaOx urolithiasis. PMID- 11911567 TI - Evaluation of Felis domesticus allergen I as a possible autoallergen in cats with eosinophilic granuloma complex. AB - OBJECTIVE: To investigate the role of Felis domesticus allergen I (Feld I) in the pathogenesis of eosinophilic granuloma complex (EGC) in cats. ANIMALS: 7 healthy cats and 6 cats with EGC. PROCEDURE: Epidermis was removed from 4 areas. Rubber stoppers filled with Feld I, saline (0.9% NaCl) solution, and PBS solution were glued to the skin lesions and removed 48 hours later. Fluid within each stopper was collected. Biopsy specimens were obtained at each site, snap frozen, and stored at -70 C. Total and differential numbers of cells in fluid were counted. Biopsy specimens were stained by use of monoclonal antibodies against feline CD4, CD8 and CD3. Data were analyzed by use of multivariate repeated-measures analysis. RESULTS: Healthy cats had a significant increase in number of CD3+ cells, compared with number of CD4+ and CD8+ cells, and Feld I caused a significant increase in number of CD3+ cells, compared with PBS or saline solutions. Cats with EGC had a significant increase in number of CD3+ cells, compared with number of CD4+ and CD8+ cells, and Feld I caused a significant increase in number of CD3+ and CD4+ cells, compared with PBS or saline solutions. Cats with EGC had an increased CD4+ response, a significantly decreased CD8+ response, and a significantly increased CD4-to-CD8 ratio compared with healthy cats. CONCLUSIONS AND CLINICAL RELEVANCE: The increased CD4+ response, significantly decreased CD8+ response, and significantly increased CD4-to-CD8 ratio are comparable to results in atopic people and allergic cats. Therefore, Feld I could be an autoallergen responsible for chronic inflammatory reactions in cats with EGC. PMID- 11911568 TI - Enteric and nasal shedding of bovine torovirus (Breda virus) in feedlot cattle. AB - OBJECTIVE: To assess fecal and nasal shedding patterns of bovine torovirus (BoTV) in cattle at time of arrival and periodically throughout the first 21 days after arrival at a feedlot. ANIMALS: 57 steers. PROCEDURE: Fecal and nasal-swab samples collected on days 0, 4, 14, and 21 after arrival were tested for BoTV, using ELISA. A subset of samples from calves testing positive and negative for BoTV was analyzed, using reverse transcriptase-polymerase chain reaction (RT-PCR). Paired serum samples were collected on days 0 and 21 and tested for BoTV antibodies, using a hemagglutination inhibition assay. RESULTS: Overall rate of fecal shedding of BoTV was 21 of 57 (37%) by ELISA and 40 of 42 (95%) by RT-PCR with peak shedding on day 4. Diarrhea was more common in calves shedding BoTV than those not shedding the virus (odds ratio, 1.72). Overall rate of nasal shedding of BoTV was 15 of 57 (26%) by ELISA and 42 of 42 (100%) by RT-PCR, with peak shedding on day 0. Specificity of the RT-PCR product was confirmed by sequence analysis. Approximately 93% of the calves seroconverted to BoTV (> 4-fold increase in titer). Differences were not detected between calves shedding BoTV and nonshedders in relation to disease and treatments, perhaps because of the low number of cattle in the study. CONCLUSIONS AND CLINICAL RELEVANCE: This study confirmed BoTV infections in feedlot cattle, including BoTV antigen and viral RNA in nasal secretions, and the shedding pattern during the first 21 days after arrival in a feedlot. PMID- 11911569 TI - Pharmacokinetics of sulfamethoxazole and trimethoprim in donkeys, mules, and horses. AB - OBJECTIVE: To compare serum disposition of sulfamethoxazole and trimethoprim after IV administration to donkeys, mules, and horses. ANIMALS: 5 donkeys, 5 mules, and 3 horses. PROCEDURE: Blood samples were collected before (time 0) and 5, 15, 30, and 45 minutes and 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 8, 10, and 24 hours after IV administration of sulfamethoxazole (12.5 mg/kg) and trimethoprim (2.5 mg/kg). Serum was analyzed in triplicate with high-performance liquid chromatography for determination of sulfamethoxazole and trimethoprim concentrations. Serum concentration-time curve for each animal was analyzed separately to estimate noncompartmental pharmacokinetic variables. RESULTS: Clearance of trimethoprim and sulfamethoxazole in donkeys was significantly faster than in mules or horses. In donkeys, mean residence time (MRT) of sulfamethoxazole (2.5 hours) was less than half the MRT in mules (6.2 hours); MRT of trimethoprim in donkeys (0.8 hours) was half that in horses (1.5 hours). Volume of distribution at steady state (Vdss) for sulfamethoxazole did not differ, but Vdss of trimethoprim was significantly greater in horses than mules or donkeys. Area under the curve for sulfamethoxazole and trimethoprim was higher in mules than in horses or donkeys. CONCLUSIONS AND CLINICAL RELEVANCE: Dosing intervals for IV administration of trimethoprim-sulfamethoxazole in horses may not be appropriate for use in donkeys or mules. Donkeys eliminate the drugs rapidly, compared with horses. Ratios of trimethoprim and sulfamethoxazole optimum for antibacterial activity are maintained for only a short duration in horses, donkeys, and mules. PMID- 11911570 TI - Dynamics of porcine circovirus type 2 infection in a herd of pigs with postweaning multisystemic wasting syndrome. AB - OBJECTIVE: To determine the pattern of infection for porcine circovirus type 2 (PCV2) in a herd of pigs with postweaning multisystemic wasting syndrome (PMWS). ANIMALS: 29 sows and 250 pigs. PROCEDURE: Blood samples were collected from all 3 , 7-, and 12-week old pigs and 59 pigs at 28 weeks of age. Pigs that died during the study were necropsied. Porcine parvovirus and PCV2 antibodies were assayed. A polymerase chain reaction (PCR) was used to detect PCV2 genome in serum of selected pigs. RESULTS: The PMWS started when pigs were 8 weeks old, with a prevalence of 30% in 8- to 10-week-old pigs. Eighty-three pigs died during the period between 3 and 12 weeks of age. Microscopic lesions consistent with PMWS were observed, and PCV2 nucleic acid was detected (50 of 68 pigs). Antibodies to PCV2 decreased from 3 to 7 weeks of age, increased at 12 weeks of age, and were maintained until 28 weeks of age. One sow had a positive result for PCR of serum. Nine, 37 and 8 pigs had PCV2 genome in serum obtained at 7, 12, and 28 weeks of age, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Infection with PCV2 coincided with severe clinical signs; however, infected 28-week-old pigs did not have evidence of disease. Immunity declined over time in young pigs. A long duration of PCV2 viremia was apparent in a high percentage of infected pigs, which may affect transmission and persistence of the virus in a herd. PMID- 11911571 TI - Response to Malassezia pachydermatis by peripheral blood mononuclear cells from clinically normal and atopic dogs. AB - OBJECTIVE: To investigate the potential cell-mediated immune response of atopic dogs to the yeast Malassezia pachydermatis and to correlate it with the type-1 hypersensitivity (humoral) response of the same population of dogs. ANIMALS: 16 clinically normal dogs, 15 atopic dogs with Malassezia dermatitis, 5 atopic dogs with Malassezia otitis, and 7 atopic control (ie, without Malassezia dermatitis or otitis) dogs. PROCEDURE: A crude extract of M pachydermatis was extracted for use as an intradermal allergy testing reagent and for stimulation of isolated peripheral blood mononuclear cells in vitro. Flow cytometry was also used to assess cell surface antigenic determinants (CD3, CD4, CD8, CD14, CD21, CD45RA, surface immunoglobulin) on peripheral blood mononuclear cells. RESULTS: Atopic dogs with cytologic evidence of Malassezia dermatitis had an increased lymphocyte blastogenic response to crude M pachydermatis extract, compared with clinically normal dogs and dogs with Malassezia otitis. Atopic control dogs did not differ significantly in their responses from atopic dogs with Malassezia dermatitis or otitis. A significant correlation was not found between the lymphocyte blastogenic response and the type-1 hypersensitivity response to M pachydermatis within any of the groups. CONCLUSIONS AND CLINICAL RELEVANCE: Cell-mediated and humoral reactivities to M pachydermatis contribute to the pathogenesis of atopic dermatitis in dogs but are not directly correlated. Modification of the dysregulated immune response toward M pachydermatis may assist in the reduction of pathologic changes associated with an atopic dermatitis phenotype in dogs. PMID- 11911572 TI - Comparison of intraosseous or intravenous infusion for delivery of amikacin sulfate to the tibiotarsal joint of horses. AB - OBJECTIVE: To establish the route of infusion (IV or intraosseous) that results in the highest concentration of amikacin in the synovial fluid of the tibiotarsal joint and determine the duration of peak concentrations. ANIMALS: 21 horses. PROCEDURE: Regional perfusion of a limb on 15 horses was performed. Amikacin sulfate was infused into the saphenous vein or via intraosseous infusion into the distal portion of the tibia (1 g in 56 ml of lactated Ringer's solution) or proximal portion of the metatarsus (1 g of amikacin in 26 ml of lactated Ringer's solution). Amikacin concentrations were measured in sequential samples from tibiotarsal joint synovial fluid and serum. Samples were obtained immediately prior to release of the tourniquet and 0.5, 1, 4, 8, 12, and 24 hours after the tourniquet was released. Radiographic contrast material was infused into the same locations as the antibiotic perfusate to evaluate distribution in 6 other horses. RESULTS: Infusion into the saphenous vein produced the highest concentration of amikacin in the tibiotarsal joint, compared with the distal portion of the tibia (mean +/- SE, 701.8 +/- 366.8 vs 203.8 +/- 64.5 microg/ml, respectively). Use of a lower volume of diluent in the proximal portion of the metatarsus produced a peak value of 72.2 +/- 23.4 microg/ml. CONCLUSIONS AND CLINICAL RELEVANCE: For regional perfusion of the tarsus, IV infusion is preferred to intraosseous infusion, because higher concentrations are achieved in the synovial fluid, and the procedure is easier to perform. PMID- 11911573 TI - Cardiovascular, respiratory, and body temperature responses of sheep to the ergopeptides ergotamine and ergovaline. AB - OBJECTIVE: To compare the effects of the ergot alkaloid ergovaline with effects of ergotamine on blood pressure, heart rate, respiratory rate, and body temperature in conscious sheep. ANIMALS: 3 sheep with indwelling arterial catheters. PROCEDURE: Ergotamine and ergovaline were injected IV (20 nmol/kg), and their effects on arterial blood pressure, heart rate, respiratory rate and pattern, body temperature, and skeletal muscle electromyographic activity were compared with control values obtained following injections of saline (0.9% NaCI) solution or acetone. RESULTS: Both ergopeptides caused immediate and significant increases in blood pressure (50 to 75 mm Hg) without concomitant increases in heart rate. Ergovaline but not ergotamine significantly increased pulse pressure (35 mm Hg). Both ergopeptides resulted in decreased respiratory rate and increased respiratory depth within the first hour of administration. Body temperature was decreased slightly upon ergopeptide administration but continued to increase thereafter, with greater increases developing with ergovaline than with ergotamine. Increased body temperatures of 3.0 to 3.5 C were maintained for at least 10 hours. Respiratory rate was increased to rates as high as 210 to 220 breaths/min in association with hyperthermia. Ergopeptides had no effect on skeletal muscle electromyographic activity. CONCLUSIONS AND CLINICAL RELEVANCE: In sheep, ergovaline has similar effects to ergotamine on cardiovascular and pulmonary function and body temperature but is more potent. These effects are consistent with clinical signs observed in the toxicoses developed when ruminants ingest grass with high concentrations of ergopeptides. PMID- 11911575 TI - Clinical and pathologic analyses of bicipital tenosynovitis in dogs. AB - OBJECTIVE: To determine the clinical and pathologic findings in dogs with primary bicipital tenosynovitis. ANIMALS: 19 dogs with 20 shoulder joints treated surgically for bicipital tenosynovitis and 8 shoulder joints from 4 clinically normal dogs. PROCEDURE: Histologic abnormalities of tendon sheaths of the biceps brachii in affected dogs were determined by use of comparison with findings in clinically normal dogs. Specimens were graded for inflammation, fibrosis, villous hypertrophy, vascular prominence, and synovial cell proliferation. Histopathologic results were statistically evaluated for relationship with clinical findings and treatment before surgery. RESULTS: Synovial villous hypertrophy and vascular prominence were the most consistent histologic findings in 16 and 14 of 20 affected joints, respectively. Evidence of inflammation was lacking in 6 joints. Ten joints had inflammatory cell infiltration of the tendon sheath. Plasma cells and lymphocytes were the most common infiltrates; however, the type and amount of inflammatory cell infiltrate were variable. Fibrosis of the tendon sheath was seen in 8 joints, and synovial cell proliferation was seen in 11 joints. Other changes included accumulation of hemosiderin, focal calcification, osseous metaplasia, lysis of collagen, and fibrocartilaginous metaplasia. No significant relationship was detected between histopathologic findings and clinical findings or treatment before surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Inflammation was more variable than hypothesized and may not be a consistent pathophysiologic feature of bicipital tenosynovitis. In some dogs, this disease may be the result of a degenerative process rather than an inflammatory process. PMID- 11911574 TI - In vitro evaluation of the role of platelet-activating factor and interleukin-8 in Mannheimia haemolytica-induced bovine pulmonary endothelial cell injury. AB - OBJECTIVE: To develop an in vitro model of the bovine alveolar-capillary interface and to evaluate the roles of interleukin-8 (IL-8) and platelet activating factor (PAF) in neutrophil-mediated endothelial injury induced by infection with Mannheimia haemolytica. SAMPLE POPULATION: Cultured bovine pulmonary microvascular endothelial cells, freshly isolated bovine neutrophils, and monocyte-derived bovine macrophages. PROCEDURE: A coculture system was developed in which endothelial cells were grown to confluence in tissue culture inserts, neutrophils were added to the inserts, and macrophages were added to tissue culture wells. Mannheimia haemolytica-derived lipopolysaccharide (LPS) or supernatant was added to activate macrophages, and inhibitors of PAF or IL-8 were added to the insert. Endothelial cell cytotoxicity and permeability (ie, albumin leakage) and neutrophil activation (ie, adhesion, degranulation [lactoferrin expression], and superoxide production) were assessed. RESULTS: The addition of M haemolytica-derived LPS to bovine macrophages in the coculture system resulted in significant increases in endothelial cell cytotoxicity and permeability and neutrophil degranulation and adhesion. Inhibition of IL-8 reduced endothelial cell permeability and neutrophil degranulation induced by exposure to M haemolytica-derived supernatant, whereas inhibition of PAF decreased superoxide release by neutrophils. CONCLUSIONS AND CLINICAL RELEVANCE: In vitro activation of bovine macrophages by M haemolytica-derived LPS resulted in neutrophil activation and neutrophil-mediated endothelial damage. Neutrophil-mediated endothelial injury and neutrophil degranulation were, at least in part, mediated by IL8, whereas PAF promoted superoxide release by neutrophils in this in vitro system designed to mimic the in vivo events that occur during the early stages of bovine pneumonic pasteurellosis. PMID- 11911577 TI - Effects of nonesterified fatty acids and beta-hydroxybutyrate on functions of mononuclear cells obtained from ewes. AB - OBJECTIVE: To assess the effects of nonesterified fatty acids (NEFA) and beta hydroxybutyrate (BHBA) on functions of mononuclear cells obtained from ewes. ANIMALS: 6 Sardinian ewes. PROCEDURE: Mononuclear cells were cultured with concentrations of NEFA (0, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, or 2,000 micromol/L) and BHBA (0, 0.45, 0.9, 1.8, or 3.6 mmol/L). Concentrations of NEFA and BHBA were intended to mimic those of ketotic or healthy ewes, and NEFA and BHBA were tested alone and in combination. Synthesis of DNA was stimulated by use of concanavalin A (Con A) or pokeweed-mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: Synthesis of DNA stimulated by Con A and PWM was significantly inhibited by high concentrations of NEFA (> or = 250 micromol/L) or by a combination of high concentrations of NEFA (> or = 250 micromol/L) and all concentrations of BHBA (> or = 0.45 mmol/L). In contrast, DNA synthesis was not inhibited by low concentrations of NEFA (< or = 125 micromol/L) or by a combination of low concentrations of NEFA (< or = 125 micromol/L) and the lowest concentration of BHBA (0.45 mmol/L). Secretion of IgM was significantly inhibited by all concentrations of NEFA and by all combinations of NEFA and BHBA concentrations. When used alone, none of the concentrations of BHBA inhibited DNA synthesis or IgM secretion. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced immunoresponsiveness during ketosis is likely to be associated with an increase in plasma concentration of NEFA and not with an increase in plasma concentration of BH BA. PMID- 11911576 TI - Multiple-center study of reduced-concentration triamcinolone topical solution for the treatment of dogs with known or suspected allergic pruritus. AB - OBJECTIVE: To determine the efficacy of triamcinolone acetonide topical solution (TTS) in dogs for use in reduction of clinical signs of pruritic inflammatory skin diseases of a known or suspected allergic basis and to evaluate adverse effects associated with TTS administration. ANIMALS: 103 pruritic adult dogs with known or suspected allergic skin disease. PROCEDURE: Dogs were treated for 4 weeks with TTS or with vehicle solution (control dogs) in a multiple-center study. Clinical signs were scored by owners and by examining veterinarians before and after treatment. Blood samples obtained before and after treatment were subjected to routine hematologic and serum biochemical analyses. RESULTS: Treatment success, as defined by an improvement of at least 2 of 6 grades in overall clinical score, was evident in 35 of 52 (67%) TTS-treated dogs (mean improvement, 1.98) and 12 of 51 (24%) control dogs (mean improvement, 0.29). For several criteria, TTS was significantly more effective than vehicle in reducing clinical signs. Minor alterations in hematologic determinations in TTS-treated dogs were limited to slightly lower total leukocyte, lymphocyte, and eosinophil counts after treatment. Minor adverse effects were reported by owners in 6 of 52 (12%) TTS-treated and 9 of 51 (18%) control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Triamcinolone used as a spray solution at a concentration approximately one-sixth the concentration of triamcinolone topical preparations currently available for veterinary use is effective for short-term alleviation of allergic pruritus in dogs. Adverse effects are few and mild and, thus, do not preclude prolonged treatment with the solution. PMID- 11911578 TI - Influence of canine recombinant somatotropin hormone on biomechanical and biochemical properties of the medial meniscus in stifles with altered stability. AB - OBJECTIVE: To determine biomechanical and biochemical properties of the medial meniscus in a semi-stable stifle model and in clinical patients and to determine the effect of canine recombinant somatotropin hormone (STH) on those properties. ANIMALS: 22 healthy adult dogs and 12 dogs with meniscal damage secondary to cranial cruciate ligament (CCL) rupture. PROCEDURE: The CCL was transected in 15 dogs, and stifles were immediately stabilized. Implants releasing 4 mg of STH/d were placed in 7 dogs, and 8 received sham implants. Seven dogs were used as untreated controls. Force plate analysis was performed before surgery and 2, 5, and 10 weeks after surgery. After 10 weeks, dogs were euthanatized, and menisci from surgical and contralateral stifles were harvested. The torn caudal horn of the medial meniscus in dogs with CCL rupture comprised the clinical group. Creep indentation determined aggregate modulus (HA), Poisson's ratio (v), permeability (k), and percentage recovery (%R). Water content (%W), collagen content (C), sulfated glycosaminoglycan (sGAG) content, and collagen type-I (cI) and -II (cII) immunoreactivity were also determined. RESULTS: Surgical and clinical groups had lower HA, k, %R, C, sGAG, cI, and clI and higher %W than the non-surgical group. Surgical stifles with greater weight bearing had stiffer menisci than those bearing less weight. Collagen content was higher in the surgical group receiving STH than the surgical group without STH. CONCLUSIONS AND CLINICAL RELEVANCE: Acute stabilization and moderate weight bearing of the CCLdeficient stifle appear to protect stiffness of the medial meniscus. Normal appearing menisci from CCL deficient stifles can have alterations in biomechanical and biochemical properties, which may contribute to meniscal failure. PMID- 11911580 TI - Calculated forelimb flexor tendon forces in horses with experimentally induced superficial digital flexor tendinitis and the effects of application of heel wedges. AB - OBJECTIVE: To calculate forces in the flexor tendons and the influence of heel wedges in affected and contralateral (compensating) forelimbs of horses with experimentally induced unilateral tendinitis of the superficial digital flexor (SDF) tendon. ANIMALS: 5 Warmblood horses. Procedure-Ground reaction force and kinematic data were obtained during a previous study while horses were trotting before and after induction of tendinitis in 1 forelimb SDF and after application of 6 degrees heel wedges to both forehooves. Forces in the SDF, deep digital flexor (DDF), and the suspensory ligament (SL) and strain in the accessory ligament (AL) of the DDF were calculated, using an in vitro model of the distal region of the forelimb. RESULTS: After induction of tendinitis, trotting speed slowed, and forces decreased in most tendons. In the affected limb, SL force decreased more than SDF and DDF forces. In the compensating limb, SDF force increased, and the other forces decreased. After application of heel wedges, SDF force in both limbs increased but not significantly. Furthermore, there was a decrease in DDF force and AL strain. CONCLUSIONS AND CLINICAL RELEVANCE: The increase in SDF force in the compensating forelimb of horses with unilateral SDF tendinitis may explain the high secondary injury rate in this tendon. The lack of decrease of SDF force in either limb after application of heel wedges suggests that heel wedges are not beneficial in horses with SDF tendinitis. Instead, heel wedges may exacerbate the existing lesion. PMID- 11911579 TI - Evaluation of small-intestinal submucosa implants for repair of meniscal defects in dogs. AB - OBJECTIVE: To assess the effects of porcine small intestinal submucosa (SIS) implants on the healing of meniscal lesions in dogs. ANIMALS: 16 adult Greyhounds of both sexes. PROCEDURE: Unilateral osteotomy was performed at time 0 to disrupt the medial collateral ligament attachment, and two (1 cranial and 1 caudal) 4-mm circular defects were created in the avascular portion of the medial meniscus. One defect was filled with an SIS graft, and the other defect remained empty (control). Three months later, the identical procedure was performed on the contralateral limb. Three months after the second surgery, dogs were euthanatized, and meniscal tissue specimens from both stifle joints were collected for gross, histologic, biomechanical, and biochemical evaluations. RESULTS: Regenerative tissue was evident in 4 (2 SIS-implanted and 2 control) of 16 defects examined histologically. In 3 defects, this thin bridge of tissue was composed of immature haphazardly arranged fibrous connective tissue with a relatively uniform distribution of fibroblasts. Aggregate modulus, Poisson ratio, permeability, and shear modulus were not significantly different between control and SIS-implanted defects either 3 or 6 months after surgery. Hydroxyproline content also did not differ between SIS-implanted and control defects at 3 or 6 months. CONCLUSIONS AND CLINICAL RELEVANCE: Implantation of porcine SIS into experimentally induced meniscal lesions in dogs did not promote tissue regeneration. PMID- 11911581 TI - Evaluation of the bispectral index as an indicator of degree of central nervous system depression in isoflurane-anesthetized horses. AB - OBJECTIVE: To determine whether the bispectral index (BIS) can be used as an indicator of degree of CNS depression in isoflurane-anesthetized horses. ANIMALS: 10 Standardbred and 6 Norwegian cold-blooded trotter stallions admitted for routine castration. PROCEDURE: A 2-channel referential electrode configuration was used to record EEG for calculation of BIS by the EEG monitor. The BIS was calculated before (awake) and after (sedated) administration of detomidine (0.01 mg/kg of body weight, IV) and butorphanol (0.01 mg/kg, IV). Anesthesia was induced with ketamine hydrochloride (2.5 mg/kg, IV) and diazepam (0.04 mg/kg, IV) and maintained with isoflurane delivered in oxygen. The BIS was calculated after 30 minutes of equilibration at an end-tidal isoflurane concentration of 1.4% (n = 8) or 1.9% (8) and recorded continuously during surgery. RESULTS: Bispectral index was significantly less in sedated and anesthetized horses, compared with awake horses. However, BIS was not significantly different between sedated and anesthetized horses. Mean BIS in horses anesthetized at 1.9% isoflurane was significantly greater, compared with horses anesthetized at an end-tidal concentration of 1.4%. Four horses in the 1.4% group moved during surgery, and BIS increased immediately prior to movement in 2 of these horses. CONCLUSIONS AND CLINICAL RELEVANCE: BIS is not a precise indicator of degree of CNS depression in isoflurane-anesthetized horses. Thus, determination of BIS may not be a useful technique for monitoring anesthetic depth in isoflurane-anesthetized horses. PMID- 11911582 TI - Effectiveness of a unique dihydropyridine (BAYTG 1000) for prevention of laminitis in horses. AB - OBJECTIVE: To determine whether a unique dihydropyridine (BAYTG 1000) would be beneficial in preventing laminitis in horses. ANIMALS: 16 clinically normal adult horses. PROCEDURE: 8 pairs of horses were used in a controlled double-blind study, using sex- and age-matched horses randomly assigned to treatment or control groups. Horses were subjected to carbohydrate overload to induce laminitis. Treated horses were administered BAY TG 1000 (30 mg/kg, PO, q 24 h) for 3 days. Hoof wall surface temperature (HWST) and lameness were recorded at 4 hour intervals. The HWST was adjusted on the basis of time of onset of lameness and evaluated, using a repeated-measures ANOVA. Lameness 8 hours after onset and clinical status 72 hours after onset of lameness were evaluated, using Mann Whitney procedures. RESULTS: Analysis revealed that BAYTG 1000 did not decrease the incidence of lameness but significantly ameliorated prodromal hypothermia, lessened the severity of lameness 8 hours after onset of lameness, and improved the clinical status of horses 72 hours after onset of lameness. CONCLUSION AND CLINICAL RELEVANCE: Results support the conclusion that BAYTG 1000 was protective when used in prevention of laminitis. The drug decreased severity and improved clinical status (recovery) of induced lameness, which was interpreted to mean that the drug's actions were on mechanisms important but secondary to primary causal mechanisms of laminitis. Therefore, drugs that enhance digital perfusion via alteration of rheologic activity may have potential use in the prevention and management of laminitis in horses. PMID- 11911583 TI - Apoptosis of bovine neutrophils during mastitis experimentally induced with Escherichia coli or endotoxin. AB - OBJECTIVE: To determine whether apoptosis of neutrophils was accelerated during mastits experimentally induced by use of Escherichia coli or E coli endotoxin and whether differences were apparent in the response to E coli or endotoxin. ANIMALS: 11 healthy lactating Holstein cows. PROCEDURE: Blood samples were collected from cows at various intervals after intramammary inoculation with E coli or endotoxin. Percentage of apoptotic neutrophils detected after in vitro incubation for 3 hours was determined. Fluorescein isothiocyanate-labeled annexin V in combination with propidium iodide was used to distinguish apoptosis and necrosis of neutrophils. Total and differential circulating leukocyte counts and rectal temperature were determined at the time of collection of blood samples. Milk yield and milk somatic cell counts were determined at the time of milking. RESULTS: Inoculation of endotoxin did not accelerate in vitro induction of neutrophil apoptosis. However, inoculation of E coli increased the percentage of apoptotic neutrophils. At 18 hours after inoculation, 20% of the neutrophils were apoptotic, compared with 5% before inoculation. Milk somatic cell count and rectal temperature increased, milk production and total leukocyte count decreased, and percentage of immature neutrophils increased after inoculation with E coli or endotoxin. However, kinetics of the responses were more rapid, more severe, and of shorter duration during endotoxin-induced mastitis. CONCLUSIONS AND CLINICAL RELEVANCE: In vitro induction of apoptosis of neutrophils was accelerated only during E coli-induced mastitis and not during endotoxin-induced mastitis. Endotoxin inoculation as a model for studying coliform mastitis in dairy cows should be viewed with caution. PMID- 11911584 TI - Plasma concentrations of endothelin-like immunoreactivity in healthy horses and horses with naturally acquired gastrointestinal tract disorders. AB - OBJECTIVE: To compare plasma endothelin (ET)- like immunoreactivity between healthy horses and those with naturally acquired gastrointestinal tract disorders. ANIMALS: 29 healthy horses and 142 horses with gastrointestinal tract disorders. PROCEDURE: Blood samples were collected from healthy horses and from horses with gastrointestinal tract disorders prior to treatment. Magnitude and duration of abnormal clinical signs were recorded, and clinical variables were assessed via thorough physical examinations. Plasma concentrations of ET-like immunoreactivity were measured by use of a radioimmunoassay for human endothelin 1, and CBC and plasma biochemical analyses were performed. RESULTS: Plasma ET like immunoreactivity concentration was significantly increased in horses with gastrointestinal tract disorders, compared with healthy horses. Median plasma concentration of ET-like immunoreactivity was 1.80 pg/ml (range, 1.09 to 3.2 pg/ml) in healthy horses. Plasma ET-like immunoreactivity was greatest in horses with strangulating large-intestinal obstruction (median, 10.02 pg/ml; range, 3.8 to 22.62 pg/ml), peritonitis (9.19 pg/ml; 789 to 25.83 pg/ml), and enterocolitis (8.89 pg/mI; 6.30 to 18.36 pg/ml). Concentration of ET-like immunoreactivity was significantly associated with survival, PCV, and duration of signs of pain. However, correlations for associations with PCV and duration of pain were low. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with gastrointestinal tract disorders have increased plasma concentrations of ET-like immunoreactivity, compared with healthy horses. The greatest values were detected in horses with large-intestinal strangulating obstructions, peritonitis, and enterocolitis. This suggests a potential involvement of ET in the pathogenesis of certain gastrointestinal tract disorders in horses. PMID- 11911585 TI - Effects of pentoxifylline on pulmonary function and results of cytologic examination of bronchoalveolar lavage fluid in horses with recurrent airway obstruction. AB - OBJECTIVES: To determine the effects of pentoxifylline (PTX) administration on lung function and results of cytologic examination of bronchoalveolar lavage fluid in horses affected by recurrent airway obstruction (RAO). ANIMALS: 10 RAO affected horses. PROCEDURES: 6 horses were orally administered PTX (16 g) mixed with corn syrup, and 4 horses were administered corn syrup alone, twice daily for 14 days. Pulmonary function was evaluated before administration (day 0) and on days 8 and 15. Bronchoalveolar lavage (BAL) was performed on days 0 and 15. Reversibility of airway obstruction was assessed by measuring pulmonary function before and after administration of atropine (0.02 mg/kg, IV). Serum concentration of PTX was measured in 4 horses 30 minutes and 2 and 4 hours after administration of PTX on days 1, 2, 3, 7 and 14. RESULTS: Administration of PTX to BAO-affected horses resulted in a decrease in elastance value on day 8 and on elastance and resistance (RL) values on days 8 and 15. Results for cytologic examination of BAL fluid obtained on day 15 did not differ significantly, compared with values for day 0. Values of RL decreased in all horses following administration of atropine. When mixed in corn syrup and administered orally, PTX was poorly absorbed in horses, and there was noticeable variation in serum PTX concentrations over time and among horses. CONCLUSIONS AND CLINICAL RELEVANCE: Based on these results, it can be concluded that administration of PTX at high doses improved respiratory function of RAO-affected horses maintained in an unfavorable environment. PMID- 11911586 TI - Amplification of 'variola virus-specific' sequences in German cowpox virus isolates. AB - In 1995 a polymerase chain reaction (PCR) protocol describing the specific detection of variola virus, the causative agent of smallpox, was published by Knight and others. Virulent variola major strains could be differentiated from less virulent variola minor strains because of the distinct amplicon sizes. Here, we applied this PCR protocol to DNA from various orthopoxvirus isolates. There was no amplification with the orthopoxvirus species vaccinia, monkeypox, mousepox, or camelpox viruses. However, amplification was observed in six out of 15 cowpox virus strains investigated. The size of the amplicons corresponded exactly with the size described for variola minor strains and the nucleotide sequence identity accounted for 97%. Findings are discussed with respect to the evolution of orthopoxvirus species assuming that variola virus most probably stems from a rodent-transmitted cowpox virus-like progenitor. PMID- 11911587 TI - Development of a monoclonal blocking ELISA for the detection of antibodies against fowlpox virus. AB - To provide a fast and easy method to detect antibodies against fowlpox virus (FWPV) particularly in high numbers of chicken sera we established a monoclonal blocking enzyme-linked immunosorbent assay (ELISA). We chose two different monoclonal antibodies (mAb), anti-FWPV 3D9/2B3 and anti-FWPV 8F3/2E11, which are both directed against the 39-kDa protein of FWPV strain HP-1. The blocking ELISA depends on the blocking of mAb binding to solid-phase antigen in the presence of positive serum. For an epidemiological study a total of 184 serum samples from Gambian chicken flocks were analysed against each of the mAbs. Four of the sera were shown to contain FWPV antibodies. These four sera showed a positive cut-off value of more than 50% inhibition exclusively in the test against the mAb anti FWPV 8F3/2E11. This phenomenon can be explained by the binding of the mAbs to distinct epitopes on the same protein. PMID- 11911588 TI - The epizootic haematopoietic necrosis virus (Iridoviridae) induces apoptosis in vitro. AB - The epizootic haematopoietic necrosis virus (EHNV) is an iridovirus causing severe disease in different fish species. We investigated the induction of apoptosis during EHNV infection of the epithelioma carp papulosum (EPC) cell line. Apoptosis reveals several characteristic morphological changes, such as chromatin condensation, nuclear fragmentation, cytoplasm membrane disorientation, or mitochondrial changes. During EHNV infection of EPC cells the occurrence of apoptosis was analysed using a fluorescein-isothiocyanate (FITC) conjugate of annexin-V to detect phosphatidylserines that have changed cytoplasm membrane localization. Annexin-V labelling was obvious 12 h after infection. At 54 h after EHNV infection 39% of the investigated EPC cells exhibited fluorescence. Furthermore, EHNV-infected cells were stained with 4'-6'-diamidino-2-phenylindole (DAPI) to detect pycnotic nuclei. Appearance of DAPI-positive nuclei was found beginning at 18 h after infection. At 54 h after EHNV infection approximately 56% of the EPC cells showed fragmented nuclei. Assays to inhibit a protein kinase dependent (e.g. double-stranded RNA-dependent protein kinase) apoptosis pathway with 2-aminopurine revealed a reduction of EHNV titres, e.g. titres were reduced 1000-fold in the presence of 100 and 200 mM 2-aminopurine. Apoptosis takes place during iridovirus infection in vitro and it seems to involve the activation of protein kinases. PMID- 11911589 TI - Recent developments in the epidemiology of virus diseases. AB - There is continual variation in viral epidemics regarding clinical symptoms, duration and disappearance, and the emergence of new diseases. This can be observed in both human and animal diseases. This evolution of virus diseases is mainly related to three factors: aetiological agent, host and environment. As far as genetic alterations of the virus are concerned, two major mechanisms are involved: mutations such as recombination and reassortment; and selection for resistance or susceptibility. This review focuses on the epidemiology of newly emerged virus diseases in man and animals, such as acquired immunodeficiency syndrome, haemorraghic fevers, bovine spongiform encephalopathy, canine haemorraghic disease and respiratory syndrome in horses. PMID- 11911590 TI - Cloning of the genomes of equine herpesvirus type 1 (EHV-1) strains KyA and racL11 as bacterial artificial chromosomes (BAC). AB - The genome of equine herpesvirus type 1 (EHV-1) strain RacL11, a highly virulent isolate obtained from an aborted foal, and that of the modified live vaccine strain KyA, were cloned as bacterial artificial chromosomes (BAC) in Eseherichia coli. Mini F plasmid sequences were inserted into the viral genomes by homologous recombination instead of the gene 71 (EUS4) open reading frame after co transfection of viral DNA and recombinant plasmid pdelta71-pHA2 into RK13 cells. After isolation of recombinant viruses by three rounds of plaque purification, viral DNA was isolated from RK13 cells infected with RacL11 or KyA virus mutants expressing the green fluorescent protein (GFP), and electroporated into Escherichia coli DH10B cells. Several bacterial colonies were shown to contain high-molecular weight BAC DNA with a restriction enzyme fragment pattern indicative of the presence of full-length RacL11 or KyA genomes. Two selected BAC clones were analysed by restriction enzyme analysis and Southern blotting, and were eventually termed pRacLI I and pKyA. respectively. Upon transfection of pRacL11 or pKyA DNA into RK13 cells, GFP-expressing fluorescing virus plaques could be identified from day 1 after transfection. Infectivity after transfection of pRacL11 or pKyA could be readily propagated on RK13 or equine cells, indicating that infectious full-length DNA clones of strains RacL11 and KyA were successfully cloned in Escherichia coli as BACs. The glycoprotein 2-negative progeny reconstituted from pRacL11 and pKyA (L11deltagp2 and KyAdeltagp2) exhibited different growth properties. Whereas both L11deltagp2 and KyAdeltagp2 extracellular titres were reduced by 15- to 32-fold, plaque diameters were only markedly (50%) reduced in the case of KyAdeltagp2. PMID- 11911591 TI - Equine herpesvirus type 1 (EHV-1) myeloencephalopathy: a case report. AB - An outbreak of neurological disease occurred in a well-managed riding school. Ataxia and paresis were observed in several horses, five of which became recumbent and were euthanized. Post-mortem analysis revealed scattered haemorrhages along the spinal cord, that were reflected by multiple haemorrhagic foci on formalin-fixed sections, with the thoracic and lumbar segments being the most affected. Pathohistologically, perivascular mononuclear cuffing and axonal swelling, especially in the white matter, were evident. Parallel to the course of disease, alterations in myelin sheets and activation of astrocytes and microglial cells were also observed. Virological findings confirmed an acute equine herpesvirus type 1 infection and virus was isolated from the spinal cord of a 26 year-old mare. PMID- 11911592 TI - Phylogenetic analysis of the 5'-untranslated region of german BVDV type II isolates. AB - On the basis of genetic differences, bovine viral diarrhoea viruses (BVDV) are subclassified into two distinct genotypes, BVDV type I and BVDV type II. We selected German BVDV type II isolates using the BVDV type I-specific monoclonal antibody WB160 and flow cytometric analysis for further characterization. For molecular characterization, a 288-bp fragment of the 5'-untranslated region (5' UTR) of the selected isolates was investigated by reverse transcription polymerase chain reaction (RT-PCR) and sequencing. Sequence comparisons of the partial 5'-UTR sequences and their phylogenetic analyses demonstrated that the 18 German BVDV type II isolates all belong to either subtype IIa (10 isolates) or subtype IIc (eight isolates). Nevertheless, the German BVDV type II isolates were genetically different (89.9-94.3% sequence identity) from the standard BVDV type II strain 890 from North America, which was recently classified as BVDV type IIa. Furthermore, a clear subdivision of the German BVDV type II isolates into two distinct subtypes (BVDV IIa Germany and BVDV IIc Germany) is shown. Viruses of both subgroups differed in the analysed 5'-UTR fragment from each other (91.6 95.2% sequence identity), but were highly conserved within the same German subtype (97.2-100% sequence identity). These findings are discussed in the context of BVDV type II origin, possible introduction into Germany, its epidemiology and impact for vaccine development. PMID- 11911593 TI - Specific detection of chikungunya virus using a RT-PCR/nested PCR combination. AB - Chikungunya (CHIK) virus is enzootic in many countries in Asia and throughout tropical Africa. In Asia the virus is transmitted from primates to humans almost exclusively by Aedes aegypti, while various aedine mosquito species are responsible for human infections in Africa. The clinical picture is characterized by a sudden onset of fever, rash and severe pain in the joints which may persist in a small proportion of cases. Although not listed as a haemorrhagic fever virus, illness caused by CHIK virus can be confused with diseases such as dengue or yellow fever, based on the similarity of the symptoms. Thus, laboratory confirmation of suspected cases is required to launch control measures during an epidemic. CHIK virus diagnosis based on virus isolation is very sensitive, yet requires at least a week in conjunction with virus identification using monovalent sera. We developed a reverse transcription-polymerase chain reaction (RT-PCR) assay which amplifies a 427-bp fragment of the E2 gene. Specificity was confirmed by testing representative strains of all known alphavirus species. To verify further the viral origin of the amplicon and to enhance sensitivity, a nested PCR was performed subsequently. This RT-PCR/nested PCR combination was able to amplify a CHIK virus-specific 172-bp amplicon from a sample containing as few as 10 genome equivalents. This assay was successfully applied to four CHIK virus isolates from Asia and Africa as well as to a vaccine strain developed by USAMRIID. Our method can be completed in less than two working days and may serve as a sensitive alternative in CHIK virus diagnosis. PMID- 11911594 TI - Mapping neutralizing and non-neutralizing epitopes on the capsid protein of feline calicivirus. AB - Neutralizing epitopes on feline calicivirus (FCV) capsid protein were mapped using chimeric capsid proteins recombinant between two FCV isolates that do not show any cross-neutralization. The three chimeric proteins examined were expressed in murine L929 cells employing an MVA/T7 vaccinia virus expression system and inoculated into major histocompatibility complex haplotype-matched C3/HN mice. Based on the neutralizing antibody titre the neutralizing epitope(s) could be mapped to the 5' hypervariable region of the E region or potentially to the C region. The epitopes of some non-neutralizing antibodies were mapped with the same chimeric proteins to the regions B or D and F of the FCV capsid protein. PMID- 11911595 TI - Escherichia coli isolates from young calves in Bavaria: in vitro susceptibilities to 14 anti-microbial agents. AB - During the occasional testing of Escherichia coli from faecal samples of young calves we observed multi-resistant isolates. Because of the significance of E. coli as an indicator bacterium for resistance trends we tested E. coli populations of young calves over a longer period. Here we present the results of a retrospective study comparing isolates from 1998 to 2000. Moreover, we compared, in a clinical study, the resistance rates of E. coli populations from 67 hospitalized calves both before and after hospitalization (with or without anti-microbial therapy), and with their anamnestic data of antibiotic usage. The highest resistance rates were found to be more than 80% for tetracyclines, ampicillin, sulfonamide/trimethoprim combinations, and chloramphenicol. A significant increase or decrease over the years was not observed. In analysing the data of hospitalized calves, an increase of resistance to some anti microbials had to be registered that seemed to be connected with the selective pressure due to agents used in the clinic. In comparing anamnestic data and resistance rates it became obvious that reliable data are not easily available and that a number of potential anti-microbial influence factors have to be taken into account. PMID- 11911596 TI - Parapoxviruses: from the lesion to the viral genome. AB - Viruses of the genus parapoxvirus from the family poxviridae cause widespread but localized diseases of small and large ruminants. The economically most important disease is contagious pustular dermatitis or contagious ecthyma among sheep and goats, often simply called orf. The parapoxviruses (PPV) can be transmitted to man leading to localized lesions that are named pseudocowpox or milkers' node as being mostly restricted to the hands and fingers. In cattle two forms of PPV manifestation are commonly observed, the bovine papular stomatitis in young calves and the occurrence of lesions at the udder of cows. We here report about the recent efforts in molecular characterization of orf viruses and the state of the art about the generation of orf virus recombinants. In addition the current knowledge on immune responses against orf viruses and some new data on the behaviour of orf virus recombinants under non-permissive conditions are reported. PMID- 11911597 TI - Documentation and evaluation of adverse drug reactions (ADR)--contribution from a poison information center. AB - The Department of Clinical Pharmacology in Jena is a pharmacovigilance center in a study on intensified spontaneous adverse drug reaction reporting. Physicians specialized in clinical pharmacology screen admissions to the Department of Internal Medicine for possible adverse drug reactions. Because of the collaboration between the Pharmacology Department and the nearby Poison Information Center (PIC) in Erfurt the question occurred whether the latter might contribute to adverse drug reaction monitoring. We compared the ADR registered by the intensified spontaneous reporting system in 1999 with those of the PIC during the same period. Each symptom observed was regarded as 1 case. Every suspected drug was also treated separately. The symptoms were classified using adverse reaction terminology. The drugs were classified according to the WHO Anatomical Therapeutic Chemical (ATC) classification index. The causality assessment was based upon bibliographic data and the method of Begaud et al. [1985]. Only possible, probable or very probable ADR were compared. The PIC registered mainly psychiatric and nervous system disorders sedation and extrapyramidal disorders were the most frequent reactions - unlike the pharmacovigilance study which registered primarily gastrointestinal and heart rate disorders. The PIC registered mainly drugs used in the therapy of disorders of the central nervous system, i.e. mostly psycholeptics and drugs acting on the alimentary tract, mostly anticholinergics. Drugs for the therapy of sensory organs disorders were frequent owing to the systemic and local adverse drug effects of anticholinergic mydriatics. The PIC and pharmacovigilance centers can benefit from co-operation. The PIC provides easy access to qualified drug information and is thus a useful tool in ADR evaluation. Although the number of adverse reactions assessed was small, their evaluation revealed problems in drug usage which would not otherwise be reported. The evaluation has enlarged the pool of ADR data which is the basis for signal detection. PMID- 11911598 TI - Severe hypoglycemia in an elderly patient treated with metformin. AB - The following case of severe hypoglycemia was reported during a systematic evaluation of hospital admissions caused by adverse drug reactions (supported by BfArM). HISTORY AND FINDINGS ON ADMISSION: A 79-year-old diabetic woman was admitted to hospital in a stuporous and unresponsive state. The initial physical examination revealed no other abnormal findings. Serum blood glucose was found to be 2.0 mmol/l and HbA1c was 4.6%. The patient had been started on antidiabetic therapy with metformin 2 months earlier. Treatment with other drugs being taken at that time, an ACE inhibitor, an NSAID and nitrofurantoin, remained unchanged. DIAGNOSIS, TREATMENT AND FOLLOW-UP: Laboratory tests excluded lactic acidosis and renal insufficiency. Cerebral computed tomography findings were normal. The patient improved dramatically following administration of glucose. Other laboratory findings confirmed the diagnosis of hypoglycemia. Blood glucose concentrations ranged between 4.0 and 10.0 mmol/l in the subsequent days and the patient could be discharged in full health. CONCLUSIONS: Drug-induced hypoglycemia is possible even in diabetics not receiving insulin or oral antidiabetic agents increasing insulin secretion. The risk of drug-induced hypoglycemia should be particularly considered when drugs containing blood glucose-lowering components are combined. Metformin does not usually cause hypoglycemia when administered as monotherapy. We suspected that hypoglycemia in this patient was caused by additional blood glucose-lowering effects of the ACE inhibitor and the NSAID possibly combined with a suboptimal nutrition. The indications for metformin administration undergo critical scrutiny. PMID- 11911599 TI - Results of systematic screening for serious gastrointestinal bleeding associated with NSAIDs in Rostock hospitals. AB - OBJECTIVES: Gastrointestinal bleeding and ulcers may lead to life-threatening complications. One of the causes is use of non-steroidal anti-inflammatory drugs (NSAIDs). METHODS: All hospital admissions in 1998 to the Departments of Internal Medicine, including the Intensive Care Unit and the Department of Surgery, in 2 hospitals in Rostock were prospectively screened for gastrointestinal bleeding. Whether the bleeding was due to an adverse drug reaction ADR or not was assessed using the rating scale of Begaud et al. [1985] for each drug taken. The risk profile and the drug history of all patients with gastrointestinal bleeding were registered. RESULTS: A total of 58 patients with gastrointestinal bleeding due to NSAIDs were documented. Risk factors for bleeding were cardiac diseases, hypertension, diabetes, age over 60 years, history of ulcer, a Helicobacter pylori infection, smoking and consumption of alcohol together with drugs known to have a risk of causing gastrointestinal bleeding and ulcers (antiplatelet drugs, anticoagulants, corticosteroids). About 70% of these patients had 3 or more risk factors, but only 20% had been receiving effective prophylaxis with a proton pump inhibitor. CONCLUSION: Gastrointestinal problems resulting from the use of NSAIDs are clinically important. It is concluded, that individual risk profiles, as a criterion for the prophylactic use of effective protective drugs, would be helpful in patients management. PMID- 11911600 TI - Results of a systematic adverse drug reaction (ADR)-screening concerning bradycardia caused by drug interactions in departments of internal medicine in Rostock. AB - OBJECTIVES: The concomitant intake of drugs, which is frequently needed, may be associated with drug interactions. We report results on the screening of ADRs responsible for hospital admissions involving bradycardia. This investigation was part of a BfArM pilot project with the objective of monitoring and reporting ADRs. METHOD: Beginning in 1997, a trained medical staff member of the Pharmacovigilance Center, Rostock, prospectively screened all hospital admissions to the Departments of Internal Medicine of the 2 hospitals in Rostock (40,000 hospital admissions). ADRs leading to hospital admission were registered, evaluated and reported. RESULTS: A total of 1,441 ADRs were recorded by the Pharmacovigilance Center Rostock in the period up to December 2000. 12% (n = 173) of all ADRs involve the cardiovascular system; 83 patients (5.7% of all ADRs) suffered from bradycardia. Bradycardia was the most frequent cardiovascular ADR observed. Of these patients, 88% were receiving 3 - 10 different drugs. Drugs suspected of causing bradycardia were: digitalis (n = 62), beta-blockers (n = 47), calcium channel blockers with negative chronotropic effect (n = 45), and antiarrhythmic drugs (n = 3). 54 patients had received more than I of these drugs concomitantly as outpatients, increasing the risk of drug interactions: 18 patients received digitalis + calcium channel blocker; 14 patients digitalis + beta-blocker; 7 patients beta-blocker + calcium channel blocker; 12 cases digitalis + beta-blocker + calcium channel blocker. CONCLUSION: The results show that special attention should be given to patients who receive more than 1 drug when there is a high risk of bradycardia. Drug combinations which may cause drug interactions should be avoided, especially when other equivalent therapeutic options are available. PMID- 11911601 TI - Assessment of frequencies of lifestyle factors and polymorphisms of drug metabolizing enzymes (NAT2, CYP2E1) in human hepatocellular carcinoma (HCC) patients in a department of surgical medicine--a pilot investigation. AB - The pathogenesis of human hepatocellular carcinoma (HCC) is a multistage process with the involvement of a multifactorial etiology. The role of drugs as risk factors has not been conclusively ascertained, but it appears that the use of oral contraceptives can be included. In the multifactorial etiology of human hepatocellular carcinoma (HCC), an association and interaction between genetic polymorphisms of xenobiotic metabolizing enzymes, lifestyle factors and cancer risk has been postulated. This pilot investigation examines the frequency of polymorphisms in selected genes (NAT2, CYP2E1) coding for xenobiotic metabolizing enzymes, and life-style habits (cigarette smoking, alcohol consumption) in 38 HCC patients. Genotyping of xenobiotic metabolizing enzymes was carried out using polymerase chain reaction--restriction fragment length polymorphism methods and DNA extracted from peripheral blood cells. In addition, HCC patients were interviewed with regard to their cigarette smoking habits and alcohol consumption using a standardized questionnaire. The results of this pilot investigation showed that the majority of the HCC patients smoke and consume alcohol. We found no predominance of slow acetylators (45%) or rapid acetylators (55%). 70.6% of slow acetylators were smokers. 86.5% of all patients with homozygote PstI/RsaI genotype also carried the homozygote DraI genotype, whereas 10.8% of all subjects with heterozygote PstI/RsaI genotype also carried the heterozygote DraI genotype. These genotype frequencies remain to be confirmed in a larger ethnic group. Whether polymorphisms of xenobiotic metabolizing enzymes is an important risk factor in (cigarette smoking-/alcohol consumption) HCC or not is currently being investigated in a case-control study in the same ethnic group. PMID- 11911602 TI - Treatment behavior of doctors regarding Helicobacter pylori infections. AB - OBJECTIVE: Since the detection of the gastric acid resistant bacterium Helicobacter pylori in the year 1982 there has been a fundamental change regarding the therapy of ulcers. According to expert opinion these infections should be treated and eradicated, whereby the so-called triple-therapies are considered to be the most effective ones. Whether such recommendations to eradicate Helicobacter pylori can be put to use in daily practice is an important question that is frequently asked. METHODS: All analyses described in the study here were done using mediplus, a longitudinal patient database with anonymous access to a representative and valid panel of physicians and patients within Germany. A total of more than 1,000 medical practices and over 75 million prescriptions can be analyzed in a cross- and/or longitudinal section. The longest time period per patient is more than 10 years starting in 1989 with monthly updates. RESULTS: With regard to existing recommendations, doctors overestimate their own compliance with the recommendations because only a fraction of traceable Helicobacter pylori infections are actually eradicated. Within the period of observation the therapy behavior has changed significantly in favor of the triple-therapies, but there are relevant differences between practitioners and internal specialists. DISCUSSION: Only a fraction of traceable Helicobacter pylori infections are adequately treated by doctors. The results show a very alarming situation due to the gap between "state of the art" and what is being achieved and carried out in daily practice. The potential for possible cost reductions and saving on resources is probably high. CONCLUSION: The results lead to the conclusion that in the treatment of Helicobacter pylori infections there is a potential for cost saving which is unused from the pharmacoeconomical point of view. PMID- 11911603 TI - Use of the mediplus patient database in healthcare research. AB - OBJECTIVE: Public health systems require fast and precise analyses of physicians' day-to-day diagnoses and therapy behavior regarding qualitative, safety or economical aspects. A partnership between physicians in practices and a database organization (IMS Health, Frankfurt, Germany) which has been in existence for more than 10 years has developed a procedure for documenting various types of studies with regard to diagnosis and therapy behavior. This endeavor has facilitated scientific progress by providing precise analytical information and guidelines. METHODS: The database used has the name mediplus. It is a longitudinal patient database with anonymous access to a representative and valid panel of physicians and patients in Germany. A total of more than 1,000 medical practices and over 75 million prescriptions have been documented in a cross and/or longitudinal section. The longest time period per patient is more than 10 years starting in 1989, with monthly updates. RESULTS: Analyses have been obtained detailing prescription behavior of doctors regarding diabetes therapy and enable recommendations to be made regarding the therapy of migraine and the eradication of Helicobacter pylori infections. Information is also retrievable on drug safety studies in general and the extent to which hospitals influence the prescription behavior of doctors treating patients after discharge. DISCUSSION: The mediplus patient database combines all decision relevant information on physicians, patients, diagnoses and course of therapies and thus makes possible the investigation of the courses of diseases and therapy patterns. The monthly update enables trends to be identified at an early stage. The mediplus database is an ideal instrument for the enforcement of quantitative and qualitative analyses of patient histories because it directly links the individual diagnoses with the corresponding therapies. The database is currently undergoing extension with additional specialist groups such as pediatricians, neurologists, orthopedists, urologists, ENT specialists, surgeons and pulmonologists. PMID- 11911605 TI - Data warehousing. PMID- 11911604 TI - Assessment of ADRs associated with lipid-lowering agents recorded in the Department of Internal Medicine, University Hospital, Jena. AB - Drug-related illness is an important cause of admission to hospital. Little information is available regarding the frequency of ADRs caused by antilipidemic agents classified as HMG-CoA reductase inhibitors (statins). Treatment with statins has been associated with the occurrence of myopathy or liver toxicity in case reports. Recent lipid intervention studies have involved the implementation of lipid lowering therapy with HMG-CoA reductase inhibitors in cardiovascular risk management. Since January 1997 we have been involved in a study, the aim of which was to improve the spontaneous drug information reporting system in Germany. The study was supported by the German Federal Institute for Drugs and Medical Devices, the "Bundesinstitut fur Arzneimittel und Medizinprodukte", Berlin BfArM. Between early 1997 and late 2000, as a result of this monitoring of ADRs, we analyzed all patient histories concerning therapy with statins. A total of 550 ADR patients were evaluated, (209 male, 341 female) with a mean age of 66.4 years. 27 (4.9%) of all patients had received statins (atorvastatin = 12, fluvastatin = 7, simvastatin as well as pravastatin = 3, lovastatin = 2). Only 2 of the 27 patients admitted to hospital for typical ADRs of statins such as skeletal muscle toxicity (e.g. myalgia, rhabdomyolysis) or disorders involving hepatic structure or function were receiving statins (atorvastatin). An increased risk of rhabdomyolysis has been reported in the case of several statins, following concomitant use with erythromycin, cyclosporine or itraconazole, all of which are potent inhibitors of CYP3A4 enzyme. But only 1 atorvastatin patient had received cyclosporine as a CYP3A4 inhibitor. After discontinuing medication, signs of intoxication disappeared. The antihyperlipidemic drugs available are generally safe and effective, and rate of ADRs is low if concomitant intake of other drugs and the differing pharmacokinetic profiles of the statins are considered. PMID- 11911606 TI - Data warehouse implementation with clinical pharmacokinetic/pharmacodynamic data. AB - OBJECTIVES: We have created a data warehouse for human pharmacokinetic (PK) and pharmacodynamic (PD) data generated primarily within the Clinical PK Group of the Drug Metabolism and Pharmacokinetics (DM&PK) Department of DuPont Pharmaceuticals. METHODS: Data which enters an Oracle-based LIMS directly from chromatography systems or through files from contract research organizations are accessed via SAS/PH.Kinetics, GLP-compliant data analysis software residing on individual users' workstations. Upon completion of the final PK or PD analysis, data are pushed to a predefined location. Data analyzed/created with other software (i.e., WinNonlin, NONMEM, Adapt, etc.) are added to this file repository as well. The warehouse creates views to these data and accumulates metadata on all data sources defined in the warehouse. The warehouse is managed via the SAS/Warehouse Administrator product that defines the environment, creates summarized data structures, and schedules data refresh. RESULTS: The clinical PK/PD warehouse encompasses laboratory, biometric, PK and PD data streams. Detailed logical tables for each compound are created/updated as the clinical PK/PD data warehouse is populated. The data model defined to the warehouse is based on a star schema. Summarized data structures such as multidimensional data bases (MDDB), infomarts, and datamarts are created from detail tables. Data mining and querying of highly summarized data as well as drill-down to detail data is possible via the creation of exploitation tools which front-end the warehouse data. Based on periodic refreshing of the warehouse data, these applications are able to access the most current data available and do not require a manual interface to update/populate the data store. Prototype applications have been web-enabled to facilitate their usage to varied data customers across platform and location. The warehouse also contains automated mechanisms for the construction of study data listings and SAS transport files for eventual incorporation into an electronic submission. CONCLUSIONS: This environment permits the management of online analytical processing via a single administrator once the data model and warehouse configuration have been designed. The expansion of the current environment will eventually connect data from all phases of research and development ensuring the return on investment and hopefully efficiencies in data processing unforeseen with earlier legacy systems. PMID- 11911607 TI - The epiphany of data warehousing technologies in the pharmaceutical industry. AB - The highly competitive pharmaceutical industry has seen many external changes to its landscape as companies consume each other increasing their pipelines while removing redundant functions and processes. Internally, companies have sought to streamline the discovery and development phases in an attempt to improve candidate selection and reduce the time to regulatory filing. In conjunction with efforts to screen and develop more compounds faster and more efficiently, database management systems (DBMS) have been developed for numerous groups supporting various R&D efforts. An outgrowth of DBMS evolution has been the birth of data warehousing. Often confused with DBMS, data warehousing provides a conduit for data residing across platforms, networks, and in different data structures. Through the use of metadata, the warehouse establishes connectivity of varied data stores (operational detail data, ODD) and permits identification of data ownership, location and transaction history. This evolution has closely mirrored and in some ways been driven by the electronic submission (formerly CANDA). The integration of the electronic submissions and document management with R&D data warehousing initiatives should provide a platform by which companies can address compliance with 21 CFR Part 11. Now more than ever "corporate memory" is being extended to the data itself. The when, why and how of successes and failures are constantly being probed by R&D management teams. The volume of information being generated by today's pharmaceutical companies requires mining of historical data on a routine basis. Data warehousing represents a core technology to assist in this endeavor. New initiatives in this field address the necessity of data portals through which warehouse data can be web-enabled and exploited by diverse data customers both internal and external to the company. The epiphany of data warehousing technologies within the pharmaceutical industry has begun and promises to change the way in which companies process and provide data to regulatory agencies. The improvements in drug discovery and reduction in development timelines remain to be seen but would seem to be rational if such technology is fully exploited. PMID- 11911608 TI - Methane production in rice soil is inhibited by cyanobacteria. AB - The present study was aimed at understanding the role of cyanobacteria and Azolla in methane production and oxidation in laboratory simulation experiments using soil samples from rice fields. All the seven cyanobacterial strains tested effected a significant decrease in the headspace concentration of methane in flooded soil, incubated under light. Synechocystis sp. was the most effective in retarding methane concentration by 10-20 fold over that in controls without cyanobacteria. The decrease in the headspace concentration of methane was negligible in nonsterile soil samples, inoculated with Synechocystis sp. and then incubated under dark. Moist soil cores (0-5 cm depth), collected from rice fields that had been treated with urea in combination with a cyanobacterial mixture, Azolla microphylla, or cyanobacterial mixture plus A. microphylla, effected distinctly more rapid decrease in the headspace concentration of methane added at 200 microl(-1) than did the soil cores from plots treated with urea alone (30, 60, 90 and 120 kg N ha(-1)), irrespective of the rate of chemical nitrogen applied to rice fields. Besides, soil cores from plots treated with urea alone at 60, 90 and 120 kg N ha(-1) oxidised methane more rapidly than did the core samples from plots treated with urea alone at 30kg N ha(-1). Cyanobacteria and A. microphylla, applied to flood water, appear to play a major role in mitigation of methane emission from rice fields-through enhanced methane oxidation. PMID- 11911609 TI - Application of RAPD analysis for identification of Lactococcus lactis subsp. cremoris strains isolated from artisanal cultures. AB - Randomly amplified polymorphic DNA (RAPD) was used for identification of Lactococcus lactis subsp. cremoris strains isolated 40 years ago from various dairy homemade products. Total genomic DNAs from six randomly chosen isolates and the reference strain Lactococcus lactis subsp. cremoris NIZO B64 were amplified using four different 10-mer primers. Although most RAPD fragments were common to all six isolates, a sufficient number of polymorphic fragments were also detected that allowed clear distinction of the isolates and the reference strain. The results indicate that RAPD analysis could be a useful and efficient method to distinguish Lactococcus lactis subsp. cremoris at the strain level and to detect genetic diversity. PMID- 11911610 TI - Purification and partial characterization of exopolygalacturonase I from Penicillium frequentans. AB - A polygalacturonase with a molecular mass of 74 kDa, an isoelectric point around pH 4.2 and pH--and temperature optima of 3.9 and 50 degrees C, respectively, was purified from a culture fluid of Penicillium frequentans. The enzyme was characterized as an exo-alpha-1,4-polygalacturonase (exo-PG I). Km and Vmax for sodium polypectate hydrolysis were 0.68 g/l and 596.8 U x mg(-1), respectively. The enzyme, a glycoprotein with a carbohydrate content of 81%, is probably the main pectinase of Penicillium frequentans responsible for cleaving monomer units from the non-reducing end of pectin. PMID- 11911611 TI - Isolation and characterization of the major nod factor of Bradyrhizobium japonicum strain 532C. AB - Bradyrhizobium japonicum 532C nodulates soybean effectively under cool Canadian spring conditions and is used in Canadian commercial inoculants. The major lipo chitooligosaccharide (LCO), bacteria-to-plant signal was characterized by HPLC, FAB-mass spectroscopy MALDI-TOF mass spectroscopy and revealed to be LCO Nod Bj-V (C18:1, MeFuc). This LCO is produced by type I strains of B. japonicum and is therefore unlikely to account for this strains superior ability to nodulate soybean under Canadian conditions. We also found that use of yeast extract mannitol medium gave similar results to that of Bergerson minimal medium. PMID- 11911612 TI - Isolation and regeneration of protoplasts from the yeast and mycelial form of the dimorphic zygomycete Benjaminiella poitrasii: role of chitin metabolism for morphogenesis during regeneration. AB - Experimental parameters for isolation and regeneration of protoplasts from the mycelial and yeast form cells of the dimorphic zygomycete Benjamininiella poitrasii are reported. Using a chitosanase containing preparation from Streptomyces sp. MCl we obtained protoplasts after 5 h incubation with a yield of 2+/-0.3 x 10(6) ml(-1) and 3+/-0.4 x 10(7) ml(-1) for the mycelial and yeast form, respectively. During regeneration under conditions triggering dimorphism the two morphological forms were observed after 36 h. Initially, for 10-12 h only an irregular mass was formed as a result of deregulated cell wall synthesis. Among the tested inhibitors influencing cell wall metabolism, chitin metabolism inhibitors showed distinctive effects on the regeneration of protoplasts suggesting that the respective enzymes significantly contribute to determining the morphogenesis of the dimorphic fungus B. poitrasii. PMID- 11911613 TI - Effect of lacZY-marking of the 2,4-diacetyl-phloroglucinol producing Pseudomonas fluorescens-strain 5-2/4 on its physiological performance and root colonization ability. AB - Transgenic Pseudomonas fluorescens 5-2/4 with reinforced 2,4-diacetyl phloroglucinol (phl) production had shown increased biocontrol ability towards Pythium ultimum (Pu), but inferior root colonization ability compared to its wild type 5.014. Therefore, enhanced root colonization ability of the transgenic strain by repeated inoculation and reisolation on tomato plants was suggested. As a preparation for repeated inoculation and reisolation cycles, the construction of a negative control of the transgenic strain 5-2/4 by marking with lacZY and screening for a mutant possessing qualities comparable to 5-2/4 was performed. Morphologically, colonies of all of the 11 selected mutants were similar on MLXgal medium. The root colonization ability of two of the lacZY-marked strains (mutants 1 and 10) was comparable to the parental strain. These were also able to compete with the resident microflora of tomato seedlings to the same extent as the parental strain. Five mutants were excluded due to lower growth rates on Yeast Malt, King's B Medium (KB) and 0.1 Tryptic Soy Agar (mutant 4, 5 and 8), excessive growth and higher siderophore production on KB (mutant 10) and increased protease production (mutant 2). With respect to in vitro-antagonism of Pu, no differences could be found between the target strain and mutants 1, 3, 6, 7 and 9. Examination of sole carbon source utilization of these five lacZY-marked strains revealed a significantly higher utilization of alpha-D-lactose and lactulose compared to 5-2/4. However, significant differences could be found for 51% of the utilized carbon sources. Cluster analysis showed a high degree of similarity between 5-2/4 and mutant 1 both when analyzed with and without alpha-D lactose. As mutant 1 also represented the colonization pattern most similar to the parental strain 5-2/4, it presents a presumptive subject for a negative control in the following inoculation and reisolation studies on tomato. PMID- 11911614 TI - Myxomycetes from upper Egypt. AB - The results of the first inventory of Myxomycetes from the subtropical region Upper Egypt are reported. The substrates were wood, bark of living and dead tree and leaf litter. 20 species belonging to 17 genera of Myxomycetes were identified. Wood was the best substrate for Myxomycetes colonization. Ceratiomyxa fruticulosa, Didymiun melanospermum, Licea biforis and Lycogala epidendrum were the most common species. Brief description and classification of species are provided. PMID- 11911615 TI - PCR amplification and polymorphism analysis of the intergenic spacer region of ribosomal DNA in Tuber borchii. AB - PCR amplification of the complete intergenic spacer region (IGS) of the Tuber borchii nuclear ribosomal repeat was obtained using universal primers CNL 12 and NS1rev. In order to improve amplification yield a specific primer, T1, was selected from a partial sequence of the IGS product. IGS diversity was characterized both at the intraindividual and intraspecific level. The results obtained at the intraindividual level showed 10% varying repeats on ten screened colonies, while at the intraspecific level the IGS polymorphism was evident as difference in length amplification between mycelial strains and fruit bodies: 3.5 kb and 2 kb respectively. PMID- 11911616 TI - The white-line-in-agar test is not specific for the two cultivated mushroom associated pseudomonads, Pseudomonas tolaasii and Pseudomonas "reactans". AB - A sharply defined white line in vitro forms between the pathogenic form of Pseudomonas tolaasii and another Pseudomonas bacterium, referred to as "reactans". This interaction has been considered as highly specific. However, results presented in this study rise doubt about the strict specificity of this interaction, as some other pseudomonads, associated with the cultivated mushroom Agaricus bisporus, also yielded a white line precipitate when they were streaked towards Pseudomonas tolaasii LMG 2342T. Moreover, some Finnish isolates inducing brown blotch symptoms on mushrooms like P. tolaasii(T), produced a typical white precipitate when streaked towards P. "reactans" LMG5329, even though phenotypical and genotypical features exclude these isolates from the species P. tolaasii. We propose that the white-line-in-agar (WLA) test should no longer be considered as an unequivocal diagnostic trait of P. tolaasii. PMID- 11911617 TI - Gallbladder and pancreatic disease during pregnancy. AB - Gallbladder disease and pancreatitis are two nonobstetric abdominal-related complaints presenting during pregnancy; gallbladder-related surgery in pregnancy is second only to appendectomy. Pancreatitis is seen less often but its most common cause is gallstone-related pain. The purpose of this manuscript is to review the clinical assessment and management of these disorders in pregnancy and to make nurses aware of the most current clinical options and techniques. PMID- 11911618 TI - Renal disease in pregnancy. AB - Anatomic and physiologic adaptations within the renal system during pregnancy are significant. Alterations are seen in renal blood flow and glomerular filtration, resulting in changes in normal renal laboratory values. When these normal renal adaptations are coupled with pregnancy-induced complications or preexisting renal dysfunction, the woman may demonstrate a reduction of renal function leading to an increased risk of perinatal morbidity and mortality. This article will review normal pregnancy adaptations of the renal system and discuss common pregnancy related renal complications. PMID- 11911619 TI - Asthma in pregnancy. AB - Asthma, a chronic inflammatory disease of the airway system, is the most common respiratory complication that can impact pregnancy. Various physiologic changes in gestation may variably affect this pulmonary disease. The cornerstone of management involves implementing strategies and treatments that assist in maintaining normal maternal pulmonary function, thereby preventing fetal complications. Optimum therapy can provide a favorable outcome for both mother and fetus. PMID- 11911620 TI - Multiple organ dysfunction syndrome (MODS) in the obstetric population. AB - Multiple organ dysfunction syndrome (MODS) has the potential to negatively affect obstetric outcomes of critically ill maternity patients. This pathophysiologic condition may often be indistinguishable from that which occurs during normal pregnancy. The normal adaptations of pregnancy, in their exaggerated form, may cause functional change to become dysfunctional in the maternal patient. Although pregnancy is considered a state of health, MODS is a grave condition with terminal outcomes. Regional perfusion deficits in oxygen and global defects of volume are two potential pathologic sequelae. Many general medical and obstetric causes may be identified. An exaggerated systemic inflammatory response syndrome (SIRS) precedes this patterned process of death. This article will apply current theories, assessment, and treatment practices of MODS to the obstetrical populace. PMID- 11911621 TI - Neonatal transitional physiology: a new paradigm. AB - Early clamping of the umbilical cord at birth, a practice developed without adequate evidence, causes neonatal blood volume to vary 25% to 40%. Such a massive change occurs at no other time in one's life without serious consequences, even death. Early cord clamping may impede a successful transition and contribute to hypovolemic and hypoxic damage in vulnerable newborns. The authors present a model for neonatal transition based on and driven by adequate blood volume rather than by respiratory effort to demonstrate how neonatal transition most likely occurs at a normal physiologic birth. PMID- 11911622 TI - Treatment of neonatal abstinence syndrome with breast milk containing methadone. AB - This article addresses the management of pregnant women participating in a methadone maintenance program. An approach to management of the labor of a woman on a methadone maintenance program is described along with a summary of what to anticipate at delivery and postpartum, and options for management of the infant who manifests symptoms of the neonatal abstinence syndrome. PMID- 11911623 TI - Thoracoabdominal aneurysm surgery and the risk of paraplegia: contemporary practice and future directions. AB - Thoracoabdominal aneurysm surgery is associated with a high incidence of morbidity and mortality. Spinal cord ischemia and the risks of paraparesis or paraplegia remain devastating complications. The mechanisms of spinal cord injury involve both acute ischemic injury and delayed reperfusion injury. Blood flow to the spinal cord frequently arises in the segment of the aorta requiring aortic cross clamping. As such, there is an obligate period of blood flow disruption. Multiple strategies have evolved to reduce the ischemic interval and to provide adjunct interventions to reduce the impact of the ischemia. Despite a multidisciplinary approach, a spinal cord ischemia is present in approximately 4 to 16% of patients, depending on the type of aneurysm and other comorbid diseases. Cerebral spinal fluid drainage, distal perfusion techniques, intercostal artery anastomosis, hypothermia techniques, and mechanisms of ischemic preconditioning are interventions employed to reduce the risk of paraplegia after thoracal-abdominal aortic surgery. Surgeons, anesthesiologists, and perfusionist are intimately involved in the decision making as to which interventions will be employed in a given case. Although these adjuncts have been evaluated in multiple animal and human protocols, the efficacy of each intervention when looked at in isolation remains difficult to determine fully. This is attributable, in part, to the complex mechanisms of the patient injury, the inherint risks of the surgical procedure, and the confounding effects of comorbid disease states. Nonetheless, clinicians must have comprehensive understanding of these various interventions and their application. This review serves as an overview of these various interventions with special emphasis on outcome data. PMID- 11911624 TI - The pathophysiology of cerebral arterial gas embolism. AB - Bubbles are introduced to the arterial circulation in many patients undergoing cardiac surgical procedures, and some of these distribute to the cerebral vessels. Larger bubbles may arrest in cerebral arterioles, causing ischemia and neuronal injury in the downstream territory. Smaller bubbles may redistribute through the cerebral circulation, but this is not a benign event. Their passage may cause transient ischemia and cause damage to endothelium. Margination and activation of leukocytes follows, and may cause a secondary ischemia. Although the potential of large bubbles to cause cerebral injury is not disputed, there is controversy over the significance of exposure to small bubbles in cardiac surgery. It is known that postsurgical neuropsychological deficits do correlate positively with numbers of emboli to which patients are exposed, but to date, the technology to distinguish between gaseous and particulate emboli or to size emboli accurately is not readily available. Until this technology becomes available and is applied in large studies designed to determine the importance of small bubbles, it seems prudent to take all practical steps to prevent introduction of arterial bubbles in cardiac surgery. PMID- 11911625 TI - Vacuum-assisted venous drainage: to air or not to air, that is the question. Has the bubble burst? AB - Assisted venous drainage is a recent development in cardiopulmonary bypass (CPB) and was introduced to overcome limitations in achieving adequate blood flow through small diameter cannulas used in minimally invasive surgery. The more common application, vacuum assisted venous drainage (VAVD) is now widely used in both adult and pediatric CPB. During a clinical investigation into pharmacological cerebral protection at Green Lane Hospital, we repeatedly observed evidence of emboli in the right common carotid artery following both entrainment of air into the venous line, and also, reductions in the blood level of the hard-shell venous reservior. We subsequently embarked upon a series of in vitro experiments designed to identify sources of emboli from the CPB circuit, and to evaluate the ability of CPB circuit components to remove air entrained into the venous line under conditions of both gravity and vacuum assisted venous drainage. Initial experiments revealed design features of certain hard-shell venous reservoirs that generated gaseous emboli. In further studies using adult circuits, entrainment of air into the venous line under conditions of conventional gravity venous drainage resulted in emboli distal to the arterial filter. When these studies were repeated using VAVD, arterial line emboli increased eight to tenfold. Initial experiments with a pediatric circuit showed similar findings. Cerebral emboli during CPB have been positively correlated with increasing neurocognitive deficits. The application of VAVD has been employed clinically without any significant redesign of the components of the CPB circuit. While VAVD may be efficacious in certain scenarios, a thorough understanding of its influence on CPB is essential. Advantages must be balanced against potential hazards. The safe use of VAVD necessitates refinement of perfusion techniques, judicious choice of application, and further development of the CPB circuit. PMID- 11911626 TI - Cerebral emboli during cardiopulmonary bypass: effect of perfusionist interventions and aortic cannulas. AB - Neuropsychological impairment is a very common complication of cardiopulmonary bypass (CPB). The principal cause of postoperative cognitive impairment is thought to be cerebral microemboli during CPB. We recently investigated the effects of perfusionist interventions and aortic cannulation techniques on cerebral emboli production during coronary bypass (CABG) surgery. Patients undergoing isolated CABG were monitored with continuous transcranial Doppler ultrasonography of the middle cerebral artery. Perfusionist interventions were defined as injections of drugs into the CPB circuit or acquisition of blood samples from the CPB circuit. Patients were randomized to receive either standard cannulation of the ascending aorta or cannulation of the distal aortic arch. Cerebral emboli were detected in all patients. The number of emboli per minute was markedly higher during perfusionist interventions than during other time periods. Patients with increased perfusionist interventions had worse neuropsychological outcomes. Cannulation of the distal aortic arch, with placement of the cannula tip beyond the cerebral vessels, resulted in significantly less cerebral emboli than cannulation of the ascending aorta. Perfusionist interventions are a common source of cerebral microemboli during CPB, and may contribute to postoperative neuropsychological impairment. Care should be taken to minimize the introduction of air into the bypass circuit during CPB. Provided it is performed safely, distal aortic arch cannulation is a useful technique for reducing cerebral emboli during cardiac surgery. PMID- 11911627 TI - How effective are cardiopulmonary bypass circuits at removing gaseous microemboli? AB - An association has been demonstrated between intravascular microemboli and organ injury during cardiopulmonary bypass (CPB). Air may be inadvertently introduced into the venous line during CPB resulting in the formation of gaseous microemboli (GME). We studied the ability of CPB circuits, from five different manufacturers, to remove GME originating from the introduction of air into the venous line. Using an in vitro model of adult CPB, 60 ml of air was introduced into the venous line and the progression of GME through the circuit components was monitored at 5 locations. In all circuits GME were detected in the arterial line following the introduction of air into the venous line. There was a wide variation between manufacturers in the ability of the circuit to remove GME. Air introduced into the venous line during CPB results in the formation of GME that are able to pass through all the circuit components including the arterial filter. The quantity of GME detected in the arterial line is influenced by the design of the circuit components and varies between manufacturers. Air in the venous line should be avoided and if present it must be dealt with promptly. PMID- 11911628 TI - A review of risk factors for adverse neurologic outcome after cardiac surgery. AB - Although the incidence of overt sequelae has traditionally been higher in patients undergoing isolated intracardiac procedures such as valve replacement or repair, recent studies show that the incidence of stroke for intracardiac procedures now approximates that for isolated coronary artery bypass grafting (CABG), in the range of 1 to 4%. In both intracardiac and extracardiac surgery, macroemboli (>200 microm in diameter) and microemboli (<40 microm in diameter) seem to be responsible for most neurologic complications. The risk of overt stroke is clearly increased in patients who undergo more complicated, combined procedures such as CABG plus valve replacement or CABG plus carotid endarterectomy. For isolated CABG, preoperative risk factors include advanced patient age, proximal aortic atherosclerosis, hypertension, previous stroke or transient ischemic attack, diabetes, and female gender. One area of controversy and current research concerns whether hypothermia is better than normothermia during cardiopulmonary bypass (CPB). Another debatable issue is whether CPB itself results in neurologic damage, owing to nonpulsatile perfusion, complement activation and the "inflammatory response," or a greater propensity for platelet activation and aggregation into microemboli in this setting. Strategies for preventing adverse neurologic outcome (new paradigms for managing intra-aortic plaque and controlling the cerebral reperfusion temperature) and for acute intervention (using specific cerebral protective agents) are under investigation. Further research into techniques for preventing or mitigating cerebral injury, particularly in high-risk patients, is clearly mandated. PMID- 11911629 TI - Surface charge properties and soil mobilities of mycoherbicidal spores. PMID- 11911630 TI - Preliminary evaluation of hair as a matrix for monitoring environmental exposure. PMID- 11911631 TI - Use of biomarkers in earthworms to detect use and abuse of field applications of a model organophosphate pesticide. PMID- 11911632 TI - Effects of zinc on brown fat thermal response to cold in normal and triiodothyronine-treated hypothyroid rats. PMID- 11911633 TI - In vivo inhibition of multivitamin on the formation of hemoglobin adduct in 4 aminobiphenyl-treated rat. PMID- 11911634 TI - Effects of cypermethrin on antioxidant enzyme activities and lipid peroxidation in liver and kidney of the freshwater fish, Oreochromis niloticus and Cyprinus carpio (L.). PMID- 11911635 TI - Acute toxicity of the pesticides endosulfan and ametryne to the freshwater prawn Macrobrachium rosenbergii De Man. PMID- 11911637 TI - Avoidance response of rainbow trout Oncorhynchus mykiss to heavy metal model mixtures: a comparison with acute toxicity tests. PMID- 11911636 TI - Cytotoxic and cytoprotective effects of selenium on bluegill sunfish (Lepomis macrochirus) phagocytic cells in vitro. PMID- 11911638 TI - Interactive effect of manganese, molybdenum, nickel, copper I and II, and vanadium on the freshwater alga Scenedesmus quadricauda. PMID- 11911639 TI - Ecotoxicological characterization of a disposal lagoon from a munition plant. PMID- 11911640 TI - Seasonal distributions of acid components in PM2.5 fraction of airborne particles in Zagreb air. PMID- 11911643 TI - Lindane and fenvalerate residues in blackgram (Vigna mungo). PMID- 11911642 TI - Dimethyl sulfide emission from waterlogged Chinese paddy soils. PMID- 11911641 TI - First results on the concentrations of some persistent organochlorines in the common hamster Cricetus cricetus (L.) in Saxony-Anhalt. PMID- 11911645 TI - Principal component analysis applied to the assessment of metal pollution from urban wastes in the Culiacan River estuary. PMID- 11911644 TI - Accumulation of polychlorinated dibenzo-p-dioxins, dibenzofurans, and dioxin-like PCBs in black-tailed gulls and eggs. PMID- 11911646 TI - Incorporation of cadmium by Acetabularia calyculus. PMID- 11911647 TI - Metals in fish and shrimp of the Campeche Sound, Gulf of Mexico. PMID- 11911648 TI - Total mercury in wild mushrooms and underlying soil substrate from the city of Umea and its surroundings, Sweden. PMID- 11911649 TI - 5'-RACEing across a bridging oligonucleotide. PMID- 11911650 TI - Blue-white selection step enhances the yield of SAGE concatemers. PMID- 11911651 TI - Rapid and efficient method for suspending cells for neurotransmitter uptake assays. PMID- 11911653 TI - Ribonuclease protection assay streamlined for high throughput with RNA in formamide and single-step rnase inactivation. PMID- 11911652 TI - Sensitive two-color whole-mount in situ hybridizations using digoxygenin- and dinitrophenol-labeled RNA probes. PMID- 11911654 TI - Cell sorting of formalin-treated pathogenic Mycobacterium paratuberculosis expressing GFP. AB - GFP is widely used as a molecular tool for the study of microbial pathogens. However, the manipulation of these pathogenic microorganisms poses a health threat to the laboratory worker, requiring biosafety level II or III containment. Although the GFPfluorophore is tolerant toformalin, a thorough analysis of this treatment on fluorescent output in prokaryotic systems has not been described. In addition, the analysis of microorganisms expressing GFP often depends on specialized equipment, which may not be housed in biosafety level II or III laboratories. Therefore, we sought to develop a safe and effective method for manipulating the GFP-expressing pathogenic bacterium Mycobacterium avium subsp, paratuberculosis (M. paratuberculosis) utilizing a formalin treatment that would permit the analysis of GFP fluorescence without requiring stringent biosafety containment. We demonstrate that formalin-treated M. paratuberculosis expresses 50% less fluorescence than viable cells, but this reduction is still compatible with spectrofluorometry and cell sorting. Furthermore, plasmid DNA that expresses GFP can be recovered efficiently from nonviable, sorted fluorescent cells. This approach is flexible, provides an additional margin of safety for laboratory personnel, and can be easily applied to other pathogenic microorganisms expressing GFP. PMID- 11911655 TI - Monitoring of cDNA microarray with common primer target and hybridization specificity with selected targets. AB - Academic researchers are increasingly producing and using cDNA microarrays. Their quality and hybridization specificity are crucial in determining whether the generated data are accurate and interpretable. Here, we describe two methods of monitoring microarray production, the sustainability of DNA attachment, and the specificity of hybridization. The first method consists of labeling an oligonucleotide, which is one of the primers used to amplify all cDNA probes on the array (except for beta-actin and GAPDH) with fluorescent dye and hybridize it to the cDNA microarray. Attachment of the cDNAs on the array after the hybridization procedure was monitored by visualizing fluorescent signals from the spots on the array. In the second method, two selected DNA targets, beta-actin and GAPDH, were labeled with fluorescent dye to hybridize to the cDNA array. Hence, hybridization specificity was demonstrated by obtaining fluorescent signals solely from the genes corresponding to the target. PMID- 11911656 TI - Dual-function vector for protein expression in both mammalian cells and Xenopus laevis oocytes. AB - Both Xenopus laevis oocytes and mammalian cells are widely used for heterologous expression of several classes of proteins, and membrane proteins especially, such as ion channels or receptors, have been extensively investigated in both cell types. A full characterization of a specific protein will often engage both oocytes and mammalian cells. Efficient expression of a protein in both systems have thus far only been possible by subcloning the cDNA into two different vectors because several different molecular requirements should be fulfilled to obtain a high protein level in both mammalian cells and oocytes. To address this problem, we have constructed a plasmid vector, pXOOM, that can function as a template for expression in both oocytes and mammalian cells. By including all the necessary RNA stability elements for oocyte expression in a standard mammalian expression vector, we have obtained a dual-function vector capable of supporting protein production in both Xenopus oocytes and CHO-K1 cells at an expression level equivalent to the levels obtained with vectors optimized for either oocyte or mammalian expression. Our functional studies have been performed with hERGI, KCNQ4, and Kv1.3 potassium channels. PMID- 11911657 TI - Simultaneous cycle sequencing assessment of (TG)m and Tn tract length in CFTR gene. AB - The lengths of the dinucleotide (TG)m and mononucleotide Tn repeats, both located at the intron 8/exon 9 splice acceptor site of the cystic fibrosis transmembrane conductance regulator (CFTR) gene whose mutations cause cysticfibrosis (CF), have been shown to influence the skipping of exon 9 in CFTR mRNA. This exon 9-skipped mRNA encodes a nonfunctional protein and is associated with various clinical manifestations in CF As a result of growing interest in these repeats, several assessment methods have been developed, most of which are, however, cumbersome, multi-step, and time consuming. Here, we describe a rapid methodfor the simultaneous assessment of the lengths of both (TG)m and Tn repeats, based on a nonradioactive cycle sequencing procedure that can be performed even without DNA extraction. This method determines the lengths of the (TG)m and Tn tracts of both alleles, which in our samples ranged from TG8 to TG12 in the presence of T5, T7, and T9 alleles, and also fully assesses the aplotypes. In addition, the repeats in the majority of these samples can be assessed by single-strand sequencing, with no need to sequence the other strand, thereby saving a considerable amount of time and effort. PMID- 11911658 TI - Tissue dispersion and flow cytometry for the cellular analysis of wound healing. AB - Injury induces a flux in the cellular composition of tissues as part of the wound healing response. There is no reliable and rapid method to quantify and characterize the cellular composition of the matrix-rich wound. Our aim was to develop a rapid method to quantify the cellular composition in wounds by tissue dispersion and flow cytometry. Age- and weight-matched mice were wounded on the dorsum using a 1.5 x 1.5 cm2 template, and the wounds were excised at predetermined time points. Tissues were dispersed into single-cell suspensions and labeled with antibodies to cell surfaces and intracellular antigens. Flow cytometry was performed to quantify the percentage of each cell population and cell death. We found that our tissue dispersion protocol resulted in low cell death (4%-6%) and very high yield (80-220 x 10(6) cells/g). Furthermore, cell surfaces and intracellular antigens were preserved to provide accurate identification of the different cell populations. With the appropriate modifications, this protocol is likely to be applicable for the viable retrieval and identification of cells from skin and other collagen-dense tissues. PMID- 11911659 TI - Bacterium-based heavy metal biosorbents: enhanced uptake of cadmium by E. coli expressing a metallothionein fused to beta-galactosidase. AB - We investigated the potential utility of a recombinant E. coli that expresses the human metallothionein II gene as a fusion protein with beta-galactosidase as a heavy metal biosorbent. E. coli cells expressing the metallothionein fusion demonstrated enhanced binding of Cd2+ compared to cells that lack the metallothionein. It was shown that the metallothionein fusion was capable of efficiently removing Cd2+ from solutions. Approximately 40% of the Cd2+ accumulated by the recombinant cells free in suspension was associated with the outer cell membrane, and 60% of that was present in the cytoplasm. PMID- 11911660 TI - Rapid identification of transformed wheat using a half-seed PCR assay. AB - A simple, nondestructive PCR-based screening method has been developed for identifying putative transgenic soft white winter wheat (Triticum aestivum L.) carrying the coat protein gene of wheat streak mosaic virus. Removal of the endosperm end of individual seed provided sufficient material for DNA extraction and PCR. DNA from seed is more free of the secondary, metabolites found in leaf tissue that can inhibit both PCR and restriction digests required for Southern analysis. The half-seed PCR assay has comparable accuracy to the leaf-tissue PCR assay and hence can be used as an accurate and rapid method for identifying transformed lines before planting. Germination of the remaining seed portion showed germination rates comparable to whole-seed controls. A slight delay in growth from the first-leaf through the first-tiller stage was observed in the half-seed-derived plants, as compared to plants grown from whole seed. PMID- 11911661 TI - Strategies for multiple sequence alignment. AB - We present an overview of multiple sequence alignments to outline the practical consequences for the choices among different techniques and parameters. We begin with a discussion of the scoring methods for quantifying the quality of a multiple sequence alignment, followed by a discussion of the algorithms implemented within a variety of multiple sequence alignment programs. We also discuss additional alignment details such as gap penalty and distance metrics. The paper concludes with a discussion on how to improve alignment quality and the limitations of the techniques described in this paper PMID- 11911662 TI - Genotyping of Snps in a polyploid genome by pyrosequencing. AB - Single-nucleotide polymorphisms (SNPs) are the most frequent DNA sequence variations, and they have become increasingly popular markers for association studies. Allelic discrimination of the mostly binary SNPs has been reported for diploid species, mainly the human, but not for polyploid genomes such as the agriculturally important crops. In the present study, we analyzed the applicability of pyrosequencing to genotyping SNPs in tetraploid potatoes. Out of 94 polymorphic loci tested, 76 (81%) proved to be amenable to allelic discrimination by pyrosequencing. An additional locus could be genotyped by the addition of an ssDNA binding protein to the pyrosequencing reaction. Of the remaining 17 loci, two failed because of the presence of paralogs in the genome, while in the other cases, self-annealing of the primer or template at the low reaction temperature (28 degrees C) employed in pyrosequencing rendered allelic discrimination impossible. The quantitative precision ofpyrosequencing was found to be similar to that of conventional dideoxy sequencing and single-nucleotide primer extension. Exceptfor some sequencespecific limitations, pyrosequencing appears to be an appropriate method for genotying SNPs in polyploid species because it is possible to distinguish not only between homoand heterozygosity but also between the different heterozygous states. PMID- 11911663 TI - Processing of immunoisolated pancreatic islets: implications for histological analyses of hydrated tissue. AB - Routine tissue processing is usually associated with histological artifacts as a consequence of shrinkage and distortion during dehydration required for embedding. With hydrated specimens such as lung, embryonic, and tissues in hydrophilic membranes, tissue processing can induce severe artifacts that interfere with adequate microscopic evaluation. Here we present a method for embedding hydrophilic alginate-polylysine microencapsulated pancreatic tissue that combines the absence of histological artifacts with a practical tissue processing method. We found that the glycol-methacrylate (GMA)-embedding method preserved the integrity of the encapsulated tissue better than snap-freezing or paraffin embedding, but the overall quality of the hydrophilic capsules remained poor Next, we modified the GMA method by introducing gradual dehydration to investigate whether the integrity of the sectioned capsules was better maintained by a more gradual pattern of water extraction. This modification resulted in well preserved morphological details of the hydrophilic membranes, hydrogel-cell interface, and encapsulated pancreatic tissue. Subsequent routine staining gave excellent contrast between the islet tissue and hydrophilic components, which allowed adequate quantitative histological and pathological comparisons. PMID- 11911664 TI - Cross-hybridization of closely related genes on high-density macroarrays. AB - DNA macroarrays are used in many areas of molecular biology research for applications ranging from gene discovery to gene expression profiling. As an increasing number of specialized macroarrays containing genes related by function or pathway are becoming available, a question that needs to be addressed is the level of hybridization signal specificity between highly similar genes that can be achieved. We have examined the ability of our LifeGrid macroarrays to distinguish hybridization signals between closely related genes. We determined the level of cross-hybridization among genes ranging from 52% to 94% sequence identity. Fragments of genes fromfive protein families were arrayed onto nylonfilters. Thefilters were subsequently hybridized with a 33P-labeled probe prepared from a pool of synthetic mRNA transcripts containing a representative of each protein family. We found that fragments containing sequences with up to 94% sequence identity displayed relatively little cross-hybridization. We conclude that this macroarray system is very specific and that hybridization signals from closely related genes can be reliably measured. PMID- 11911665 TI - High-speed plasmid isolation using 96-well, size-exclusion filter plates. AB - The accelerating pace of genomics analysis has necessitated the abbreviation of DNA sample preparation protocols. We have developed a size-exclusion-based system for the rapid isolation ofplasmid DNA in a 96-well microplate format. This high speed protocol employs a modified alkaline lysis methodfor the preparation of the bacterial lysate, followed by three short vacuum filtration steps. Unlike traditional bind/wash/elute methods, there is no need to use chaotropic salts or ethanol. The samples are recovered from the top side of the MultiScreen96 PLASMID plates. Starting with bacterial cell pellets, the entire prycedure for purifying the plasmid DNA can be performed in 30 min with a multichannel pipettor. The high yields, reproducibility, and quality of the plasmids make this system a good choice for any cloning or DNA sequencing operation. PMID- 11911666 TI - Protein interaction-targeted drug discovery: evaluating critical issues. AB - Employment of the decision strategies outlined in this general discussion should help to pinpoint mode of activity in drug development and validation. Overall, as a paradigm for drug development, a search for small molecules that can interfere with PPIs would seem to have significant long term potential. At present, the level of structural knowledge in databases is not sufficient to predict in toto the protein binding properties of a modeled drug, but as databases improve, this may become generally feasible. A major point that remains to be determined is how much specificity of protein binding can be incorporated into molecules of generally less than 500 Da. Finally, integration of PPI-targeting strategies with other approaches towards drug design will enhance the number of signaling pathways that can effectively be targeted. These points will be particularly pertinent as technologies permit a systematic identification of encoded protein interactions that govern the proteornic complement of cells. PMID- 11911667 TI - Markers of apoptosis: methods for elucidating the mechanism of apoptotic cell death from the nervous system. AB - Apoptosis is a highly conserved energy-requiring program for non-inflammatory cell death that is important in both normal physiology and disease. Numerous techniques have been used to study apoptosis. In the nervous system, apoptosis is necessary for normal development, but it also occurs in many acute and chronic pathologic conditions. This review places commonly used markers of apoptosis and their detection in the context of what is now known about the process of apoptosis. We review the potential role of apoptosis in nervous system and neurodegenerative disorders (Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis). We then describe important morphological, immunocytochemical, and molecular genetic markers for apoptosis, including proteases, signal transduction molecules, and mitochondrial proteins. The possibility of manipulating apoptosis therapeutically in conditions of too many or too few cells is under active investigation. PMID- 11911668 TI - Two-color, fluorescence-based microplate assay for apoptosis detection. PMID- 11911669 TI - Robotic high-throughput assay for isolating apoptosis-inducing genes. AB - The need to assign functions to genes will increase in upcoming years because of the accumulation of sequence information from various organisms. Here we present a genetic screen for the systematic functional analysis of genes in mammalian cells. The screen is based on high-throughput, transient transfections of individual cDNAs by a robotic system. This construction uses a moveable pipetting head with a novel 96-channel parallel tube-pump array and an integrated de lidding device. Because of the robot design, the processing speed can reach 2,000 transfections/h. We demonstrate the suitability of the screen to isolate apoptosis-inducing genes as a first read-out. This is achieved in combination with co-transfected reporter plasmids to produce a novel and convenient assay for apoptosis induction. PMID- 11911670 TI - Use of CMFDA and CMTMR fluorescent dyes in FACS-based antibody screening. AB - Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a control cell line not expressing the antigen. Antibodies binding only to the transfected cell line are then selected for further analysis. This approach can be challenging if a large number of antibodies need to be screened and the antibody quantities are limited. Here we describe a novel method that combines the above two steps of FACS screening into a single step, based on the use of two fluorochromes, CMFDA and CMTMR, to stain transfected and control cell lines, respectively. Antibodies conjugated to a thirdfluorochrome are then added to the combined cells, followed by three-color FACS analysis. The newly modified FACS method is simple, sensitive, and high throughput. It can be used for antibody screening in multiple cell lines simultaneously. PMID- 11911671 TI - Determination of growth rate of microorganisms in broth from oxygen-sensitive fluorescence plate reader measurements. AB - A novel method utilizing the BD Oxygen Biosensor System has been developed to rapidly, simply, and accurately determine the growth rate of microorganisms in broth, with no needfor plate counts, standardized inocula, or technically difficult manipulations. The BD Oxygen Biosensor System incorporates an oxygen sensitive material into the wells of standard Falcon microplates. The time response of this sensor monitored in afluorescence plate reader can be used to quantitate microbe growth. The method entails seeding a dilution series of microorganism onto the plate and reading at regular intervals for 3-10 h. As the organisms grow and consume oxygen, thefluorescence intensities increase over time to form a family of sigmoidal growth curves. A simple mathematical analysis of the time intervals between the curves yields the doubling time, which is independent of the initial concentration of organism. The method is ideally suited as a screening tool for assessing the impact of culture conditions, media composition, or added compounds on growth kinetics. PMID- 11911672 TI - Natural cytotoxic cells of gilthead seabream: maximum percentage of lysis. AB - The maximum percentage of lysis of head-kidney non-specific cytotoxic cells (NCC) against mammalian tumour cells (L1210 and K562) in the marine teleost gilthead seabream (Sparus aurata L.) was studied. The present data indicate the short period of time necessary for gilthead seabream NCC to form conjugates and deliver a lethal hit. The maximum percentage of lysis observed demonstrates that seabream NCC activity against L1210 tumour cells is faster than against K562 tumour cells. This kinetic parameter suggests that fish NCC show a less efficient cytotoxic activity than their mammalian counterparts. The possibility of applying theoretical treatments to systems consisting of lower vertebrate non-specific cytotoxic cells and tumour targets, similar to those applied to mammals, is considered, and the phylogenetic implications of our findings are discussed. PMID- 11911673 TI - Dietary vitamin C and its derivatives affect immune responses in grass shrimp, Penaeus monodon. AB - Effects of L-ascorbic acid (AA) and its four derivatives, namely L-ascorbyl-2 sulfate (C2S), L-ascorbyl-2-polyphosphate (C2PP), L-ascorbyl-2-monophosphate-Na (C2MP-Na) and L-ascorbyl-2-monophosphate-Mg (C2MP-Mg) on the immune responses of juvenile grass shrimp, Penaeus monodon, were studied. The vitamin C deprived diet together with diets supplemented with either adequate or high (five times adequate) levels of AA, C2S, C2PP, C2MP-Na and C2MP-Mg were each fed to triplicate groups of shrimp (mean initial weight: 0.37 +/- 0.01 g) for 8 weeks. Significantly (P<0.01) higher weight gain (WG), feed efficiency (FE), survival, total haemocyte count (THC), superoxide anion (O2) production ratio and phenoloxidase (PO) activity were observed in shrimp fed diets supplemented with adequate and high levels of ascorbate than shrimp fed the vitamin C deprived diet, regardless of the ascorbate source. Among the ascorbate sources, shrimp fed C2MP-Mg and C2PP containing diets had higher THC than shrimp fed AA, C2S and C2MP Na containing diets, regardless of the supplementation level. Shrimp fed adequate levels of C2MP-Mg and C2PP and high levels of C2MP-Mg containing diets had higher O2 production ratios than shrimp fed AA and C2S containing diets. Shrimp fed adequate levels of C2MP-Mg and C2PP and high levels of C2PP containing diets had higher PO activity than shrimp fed AA, C2S and C2MP-Na containing diets. These data suggest that dietary ascorbate enhances immune responses in P. monodon and different ascorbate sources may affect the immune responses differently. PMID- 11911674 TI - In vitro manipulations of vitamin C and vitamin E concentrations alter intracellular O2- production of hybrid striped bass (Morone chrysops x Morone saxatilis) head-kidney cells. AB - To examine the mechanism by which vitamins C and E alter phagocyte function, a series of in vitro manipulations were conducted with cells isolated from the head kidney of hybrid striped bass (average weight 680 g) fed a diet supplemented with minimum requirement levels of vitamins C and E for 2 weeks. Head-kidney phagocytes were cultured in media containing physiologically deficient (23 microM, adequate (45 microM) or excessive (182 microM) concentrations of vitamin C, and physiologically deficient (5 microM), adequate (9 microM) or excessive (32 microM) concentrations of vitamin E for 18 h. Following culture and stimulation, levels of reactive oxygen intermediates and hydrogen peroxide were determined. There were no effects of vitamin C or vitamin E concentrations on hydrogen peroxide or extracellular O2- generation. Intracellular O2- production, however, was significantly (P < or = 0.05) affected. When vitamin C was supplied at deficient levels to the medium, vitamin E elevated O2- production to levels not different from those of cells incubated with requirement levels of both vitamins. Similarly, when vitamin E was deficient in the media, vitamin C supplementation at requirement levels normalised intracellular O2- production. This data provides support for the presence of a vitamin C and vitamin E sparing mechanism in phagocytic head-kidney cells of hybrid striped bass and yield some insight into the mechanisms by which vitamin C and vitamin E function in immunomodulation. PMID- 11911676 TI - Resistance to serum killing may contribute to differences in the abilities of capsulate and non-capsulated isolates of lactococcus garvieae to cause disease in rainbow trout (Oncorhynchus mykiss L.). AB - Three capsulated and two non-capsulated isolates of Lactococcus garvieae were investigated in terms of their wall proteins, virulence and interactions with rainbow trout immunoglobulin (Ig). All isolates were similar in integral membrane protein profile, and all were able to bind non-immune rainbow trout Ig, although different proteins appeared to be involved in Ig binding. However, whilst capsulated isolates were highly virulent, non-capsulated isolates were avirulent. This appeared to correlate with susceptibility of the non-capsulated isolates to rainbow trout normal serum. In contrast, the capsulated isolates were resistant to both normal and immune serum killing. In spite of this, passive immunisation of rainbow trout with specific anti-serum to L. garvieae was able to protect against challenge by capsulated isolates of L. garvieae. This suggests the antibody may have some other role in protection against disease caused by this important Gram-positive bacterial fish pathogen. PMID- 11911675 TI - Survival of Flavobacterium psychrophilum in rainbow trout (Oncorhynchus mykiss) serum in vitro. AB - Virulent and non-virulent strains of Flavobacterium psychrophilum of different serotypes were examined for survival and growth in non-immune and immune rainbow trout serum, in vitro. A majority of the examined strains consumed complement of non-immune serum, but the complement cascade was not able to cause an immediate (after 3 h incubation) notable reduction in viability of the inoculated cells. After 24 h incubation a more pronounced reduction in the number of viable bacteria was observed in untreated serum as well as in serum heated at 45 degrees C. In serum heated at 56 degrees C this reduction in cell number was not observed, but an increase in cell number did not occur either. The serum survival of one of the examined strains was different from the others in showing cell multiplication after 24 h incubation in normal as well as heat-treated (45 and 56 degrees C) serum. In immune serum no immediate reduction in viability of inoculated cells, of all tested strains, was observed. The number of viable cells showed a slow decrease or remained almost unchanged for up to 72 h post inoculation in untreated serum, at 5 degrees C as well as 15 degrees C. In heat treated serum (45 degrees C) the number of viable cells decreased slowly at 5 degrees C and 15 degrees C for up to 72 h. The results suggest that the examined strains were unaffected by the alternative complement reaction present in fish serum as well as by antibodies against F. psychrophilum. However, some unknown component(s) in the fish sera, or lack of nutrients or essential growth factors, inhibited the growth of most of the examined strains in the tested fish sera. PMID- 11911677 TI - The outer membrane fraction of Flavobacterium psychrophilum induces protective immunity in rainbow trout and ayu. AB - Flavobacterium psychrophilum is the causative agent of coldwater disease, which is responsible for serious losses in fish aquaculture in several parts of the world. No commercial vaccines are currently available for the prevention of coldwater disease. The present study sought to assess the efficacy of a F. psychrophilum vaccine based on the antigenic outer membrane fraction (OMF). This fraction induced significantly higher protection against coldwater disease in both rainbow trout (Oncorhynchus mykiss) and ayu (Plecoglossus altivelis) compared to inactivated whole cell F. psychrophilum bacterin. Coincident with higher protection, sera of fish immunised with the OMF vaccine had higher agglutination titres than those of fish immunised with inactivated whole cell F. psychrophilum. PMID- 11911678 TI - Antibody increases phagocytosis and killing of Lactococcus garvieae by rainbow trout (Oncorhynchus mykiss, L.) macrophages. AB - The present study reports that specific antibody increased the bactericidal activity of rainbow trout head-kidney macrophages against virulent capsulated Lactococcus garvieae in the absence of complement. The observed increased bactericidal activity appeared to result from increased phagocytosis of capsulated L. garvieae in the presence of specific antibody and may in part explain the protective effect of antibody previously reported against this disease. PMID- 11911679 TI - The virulence of Enterococcus to freshwater prawn Macrobrachium rosenbergii and its immune resistance under ammonia stress. AB - Growth of pathogen bacterium. Enterococcus was not affected in tryptic soy broth (TSB) medium containing ammonia-N concentration in the range of 0-5.14 mg l(-1). Giant freshwater prawn Macrobrachium rosenbergii (8-12 g) were challenged with Enterococcus which had been incubated for 24 h in TSB medium containing different concentrations of ammonia-N at 0-5.14 mg l(-1) Cumulative mortality of M. rosenbergii was higher for the bacteria incubated in TSB medium having ammonia-N at 0 and 0.26 mg l(-1), than those incubated in TSB medium having 1.28, 2.57 and 5.14 mg l(-1) ammonia-N after 24 h of challenge. However, cumulative mortality of prawn was significantly higher for the bacteria incubated in TSB medium with no ammonia added after 120 h of challenge. The prawns (8-12 g) were challenged with Enterococcus previously incubated in TSB medium for 24 h, then placed in water having concentrations of ammonia-N at control (0.06 mg l(-1)), 0.55, 1.01, 1.68 and 3.18 mg l(-1). Mortality of prawns increased directly with ammonia-N concentrations after 72 h challenge. The pranws (20-30 g) which had been exposed to control, 0.55, 1.68 and 3.18 mg (-1) ammonia-N for 7 days were examined for the total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase activity and respiratory burst of haemocytes. Phenoloxidase activity decreased when the prawns were exposed to ammonia-N greater than 0.55 mg l(-1). The respiratory burst increased significantly at 0.55 mg l(-1) but decreased significantly at 1.68 and 3.18mg (-1) ammonia-N. No significant difference in haemocyte count was observed among the prawns at different ammonia N concentrations. It is suggested that ammonia in water decreases the virulence of Enterococcus, and reduces the immune resistance of M. rosenbergii. PMID- 11911680 TI - A patent searcher's personal chronicle: 40 years in the evolution of a profession. AB - Forty years have passed since this chemist elected to attempt to become an information chemist, thus pursuing a career path he had not heard of during his undergraduate and graduate education. Much has changed during that period. Punched cards, microfilm and microfiche, coordinated term indexes, and more have come and gone. The US Patent and Trademark Office has issued more than 3 million patents, exceeding its total output in all the years that came before. Online database searching replaced prior reliance on printed indexes and classified card files, and now seeks to redefine itself to stand up against the juggernaut of the Internet. In the face of all that change the traditional abstracting and indexing function is still with us, though not without considerable reshaping. This paper surveys this landscape of change and suggests that if we are wise, we will nurture this intellectual activity far into the future. PMID- 11911681 TI - Java classes for managing chemical information and solving generalized equilibrium problems. AB - Java classes have been created for organizing chemical information and solving generalized equilibrium problems. An object-oriented approach is employed for the organization and manipulation of chemical information. Classes have been created to represent chemical species, phases, and chemical reactions. The representation of the entire chemical system is encapsulated in the ChemSystem class. The Equilibria class provides methods for analyzing a chemical system, as described by a ChemSystem object, and determining the amounts of each species in the system at equilibrium. The ChemEquilibria applet has been created to facilitate deployment of this software over the World Wide Web. PMID- 11911682 TI - Fast generation of an alkane-series dictionary ordered by side-chain complexity. AB - Selecting the main chain of an alkane as the path that yields the least complex side chains without the maximum-length constraint leads to an efficient generating algorithm representable as nested binary trees. The largest side chain required to specify an N-carbon alkane becomes (N-1)/3. This allows 3.8 million C1-C22 alkanes to be coded for name translation in dictionary order, using an alphabet of 33 C1-C6 alkyl groups also ranked by complexity. The generating process produces reversible isomer codes already in canonical order, making the computation rate in isomers per second inverse linear with N and much faster than reported rates for other structure generators. PMID- 11911683 TI - On a nonelementary progress curve equation and its application in enzyme kinetics. AB - The analytical equation describing progress curves of an enzyme catalyzed reaction acting upon the Michaelis-Menten mechanism has been known for the case in which only the free enzyme incurs a loss of its activity, either spontaneously or as a result of an irreversible inhibitor action. The solution of differential equations which defines the rates of enzyme inactivation and substrate utilization is expressed by a nonelementary function in equation of an implicit type that precludes direct calculation of the extent of reaction at any time. Previously, the implicit equations have been rearranged to the alternative formulas and solved by the Newton-Raphson method, but this procedure may fail when used upon the presented equation. For this reason the other root-finding numerical method was applied, and the enzyme kinetic parameters of such numerically solved implicit equation for the reaction mechanism of irreversibly inhibited acetylcholinesterase were fitted to the experimental data by a nonlinear regression computer program. PMID- 11911684 TI - Fuzzy ARTMAP and back-propagation neural networks based quantitative structure property relationships (QSPRs) for octanol-water partition coefficient of organic compounds. AB - Quantitative structure-property relationships (QSPRs) for estimating the logarithm octanol/water partition coefficients, logK(ow), at 25 degrees C were developed based on fuzzy ARTMAP and back-propagation neural networks using a heterogeneous set of 442 organic compounds. The set of molecular descriptors were derived from molecular connectivity indices and quantum chemical descriptors calculated from PM3 semiempirical MO-theory. Quantum chemical input descriptors include average polarizability, dipole moments, exchange energy, total electrostatic interaction energy, total two-center energy, and ionization potential. The fuzzy ARTMAP/QSPR performed, for a logK(ow) range of -1.6 to 7.9, with average absolute errors of 0.03 and 0.14 logK(ow) for the overall data and test sets, respectively. The optimal 12-11-1 back-propagation/QSPR model, for the same range of logK(ow), exhibited larger average absolute errors of 0.23 and 0.27 logK(ow) for the test and validation data sets, respectively, over the same range of logK(ow) values. The present results with the fuzzy ARTMAP-based QSPR are encouraging and suggest that high performance logK(ow) QSPR that encompasses a wider range of chemical groups could be developed, following the present approach, by training with a larger heterogeneous data set. PMID- 11911685 TI - The comparative molecular surface analysis (COMSA)--a nongrid 3D QSAR method by a coupled neural network and PLS system: predicting pK(a) values of benzoic and alkanoic acids. AB - A self-organizing neural network was used to design a novel method capable of the quantitative prediction of molecular properties. The method is based on the comparison of molecular surfaces performed by the coupled neural network and PLS system. Unlike CoMFA and related methods it does not compare the properties describing a discrete set of points but the average property values calculated for a certain area of the molecular surface. It has been found that the results of the PLS analysis of the series of the comparative matrices of the molecular electrostatic potential (MEP) are quite stable. Also the results only slightly depend on such parameters as the number of points sampled at the molecular surface (D) or a winning distance (MD) of the self-organizing neurons. The influence of these parameters for modeling the effects limited by steric and electronic effects was determined and the pK(a) values of the ortho-, meta-, and para- (o-, m-, p-) analogues of benzoic acid and selected alkanoic acids were predicted. We generally found that for the series analyzed CoMSA gave better models than CoMFA. PMID- 11911686 TI - A web-based 3D-database pharmacophore searching tool for drug discovery. AB - Three-Dimensional (3D) structural database pharmacophore searching has become a very effective approach for discovery of novel lead compounds in drug discovery. Although several commercial programs are available, these commercial programs are primarily used as a stand alone and require a local database. In recent years, the Internet has become the main medium of choice for multiuser application program distribution. Herein, we describe our development of a Web-based 3D database pharmacophore-searching tool based on the server-client Web architecture. Both rigid and conformationally flexible searching methods are implemented. Our results show that for a typical three-center rigid pharmacophore search, the run time for searching 50 000 compounds is less than three minutes, and for four-center pharmacophore searching, the run time is less than 10 minutes on a desktop computer. For a flexible 3D-pharmacophore search, the run time for searching 50 000 compounds generally takes between one and several hours. The search results are comparable to those obtained using a commercial program. We expect that this Web-based tool will be very useful for scientists who are interested in 3D-database pharmacophore searching via the Internet. PMID- 11911687 TI - SIRS-SS: a system for simulating IR/Raman spectra. 2. Procedures and performance. AB - This paper is devoted to the description of procedures used in our IR/RAMAN spectrum simulation system, based on substructure/subspectrum correlations established between linked databases. The search is performed in the following order: small molecules/specific fragments/atom centered FRELs (FREL: FRagment centered on an Environment which is Limited)/bond focused FRELs. Comparative study with several reported methods has been carried out to show good performance of this software both for IR and RAMAN spectrum simulation. PMID- 11911688 TI - MECHGEN: computer aided generation and reduction of reaction mechanisms. AB - The paper describes selection rules implemented in a software generating "possible reaction mechanism", i.e. a set of elementary reactions chosen from all stoichiometrically possible reactions. The novelty of the approach lies in the fact that the user has to define all species involved (reactants, intermediates, products), and the rules applied with user-set limits reduce the resulting mechanism to a reasonable set of possible elementary reactions. The computer code consists of five parts: (i) definition of species, and introducing its characteristics (structure and thermodynamic data); (ii) definition of the reacting system and generation of all stoichiometrically possible reactions; (iii) reduction of the mechanisms using complexity and thermodynamic constraints based on user-set limits; (iv) calculation of the resulting pathways (routes of the various atoms or groups of atoms transferred from one species to another); and (v) tools to help visualization of the process by finding those elementary processes which realize a given pathway. Reasonable flexibility is ensured for using selection rules based on various criteria with limits set by the user. The various pathways are shown (in a matrix form), which offers an overview of the entire process. PMID- 11911689 TI - Information and organic molecules: structure considerations via integer statistics. AB - Information in relation to organic molecules was investigated in a previous work (Graham and Schacht, J. Chem. Inf. Comput. Sci. 2000, 40, 187). The topic is given further consideration here with the help of integer statistics. Discussed are the ramifications of an integer variable Omega(t) which quantifies the total number of binding complexions for an organic molecule. Offered is a statistical view of the maximum allowed number of independent regions D expressed by the molecule, dependent on Omega(t). We illustrate the distribution properties of D along with upper limit estimates of the regioinformation mu, also dependent on Omega(t). Integer statistics based on elementary number theory establish the key distribution properties of D and mu. In so doing, the traits distinguishing high regioinformation molecules are enumerated. The statistical approach encompasses all possible molecules and conditions, not just those reported to date in chemical databases. The aim is to view the regioinformation expressed by molecules in an alternative and general way. PMID- 11911690 TI - A Chinese Postman Problem based on DNA computing. AB - DNA computing is a novel method for solving a class of intractable computational problems, in which the computing can grow exponentially with the problem size. Up to now, many accomplishments have been achieved to improve its performance and increase its reliability. A Chinese Postman Problem has been solved by means of molecular biology techniques in the paper. A small graph was encoded in molecules of DNA, and the "operations" of the computation were performed with standard protocols and enzymes. This work represents further evidence for the ability of DNA computing to solve NP-complete search problems. PMID- 11911691 TI - Correlation of the melting points of potential ionic liquids (imidazolium bromides and benzimidazolium bromides) using the CODESSA program. AB - The melting points of several imidazolium-based ionic liquids or ionic liquid analogues were correlated using the CODESSA program in order to develop predictive tools for determination of suitable ionic liquid salts. The data set consisted of melting point data (degrees C) for 104 substituted imidazolium bromides divided on the basis of the N-substituents into three subsets: A-57 compounds, B-29 compounds, and C-18 compounds. The 45 benzimidazolium bromides form set D. Five-parameter correlations were obtained for (i) set A with R2 = 0.7442, (ii) set B with R2 = 0.7517, and (iii) set D with R2 = 0.6899, while set C was correlated with a three parameter equation with R(2) = 0.9432. These descriptors for predicting the melting points of the imidazolium and benzimidazolium bromides were based on the size and electrostatic interactions in the cations. PMID- 11911692 TI - Prediction of glass transition temperatures from monomer and repeat unit structure using computational neural networks. AB - Quantitative structure-property relationships (QSPR) are developed to correlate glass transition temperatures and chemical structure. Both monomer and repeat unit structures are used to build several QSPR models for Parts 1 and 2 of this study, respectively. Models are developed using numerical descriptors, which encode important information about chemical structure (topological, electronic, and geometric). Multiple linear regression analysis (MLRA) and computational neural networks (CNNs) are used to generate the models after descriptor generation. Optimization routines (simulated annealing and genetic algorithm) are utilized to find information-rich subsets of descriptors for prediction. A 10 descriptor CNN model was found to be optimal in predicting T(g) values using the monomer structure (Part 1) for 165 polymers. A committee of 10 CNNs produced a training set rms error of 10.1K (r2 = 0.98) and a prediction set rms error of 21.7 K (r2 = 0.92). An 11-descriptor CNN model was developed for 251 polymers using the repeat unit structure (Part 2). A committee of CNNs produced a training set rms error of 21.1K (r2 = 0.96) and a prediction set rms error of 21.9 K (r2 = 0.96). PMID- 11911694 TI - New diversity calculations algorithms used for compound selection. AB - Some modifications were introduced into the previously described Centroid diversity sorting algorithm, which uses cosine similarity metric. The modified algorithm is suitable for the work with large databases on personal computers. For example, for diversity sorting of the database with the size greater than a million of records, less than 9 h are required (Pentium III, 800 MHz). The problem of selecting new compounds into the existing collection is examined to reach the maximum diversity of the collection. The article describes the new algorithm for the selection of heterocyclic compounds. PMID- 11911693 TI - Validation of structural proposals by substructure analysis and 13C NMR chemical shift prediction. AB - The 2D NMR-guided computer program COCON can be extremely valuable for the constitutional analysis of unknown compounds, if its results are evaluated by neural network-assisted 13C NMR chemical shift and substructure analyses. As instructive examples, data sets of four differently complex marine natural products were thoroughly investigated. As a significant step towards a true automated structure elucidation, it is shown that the primary COCON output can be safely diminished to less than 1% of its original size without losing the correct structural proposal. PMID- 11911695 TI - A new class of molecular shape descriptors. 1. Theory and properties. AB - The integrals V (n1, n2, n3) = integral dr x(n)1 y(n)2 z(n)3, where integral dr represents integration over the volume of a body, such as a molecule, where x, y, and z are Cartesian coordinates of a point in the interior of the body relative to an arbitrary reference frame, and where n1, n2, and n3 are integers greater than or equal to zero, constitute moments of the volume distribution of the body. Considering all such quantities for which 0 < or = n1 + n2 + n3 < or = 6 gives a set of 84 independent numbers which characterize the shape of the body and constitute a very useful set of shape descriptors. They also carry information about the absolute orientation and position of the body, and because their behavior under rotations and translations can be calculated quickly, they provide a fast, robust algorithm for the alignment of two similar molecules as well as a qualitative measure of their similarity. This paper reports the performance of the alignment algorithm on a learning set of about 80 different shapes. The algorithm is further tested against a set of small drug-like compounds that have been screened as anticancer agents. In both cases, excellent alignments of "shape similar" molecules are obtained. Discussions are provided on many basic properties of these moments, e.g., their behavior under translations and rotations of the reference frame and their symmetry properties. PMID- 11911696 TI - Approximate derivative calculated by using continuous wavelet transform. AB - A novel method of calculating approximate derivative of signals in analytical chemistry by using the continuous wavelet transform (CWT) is proposed. As compared with numerical differentiation, FT method and DWT method, fast calculation, and simple mathematical operation are remarkable advantages of CWT method. The signal-to-noise ratio (SNR) of approximate derivative of signals calculated by CWT method is easily enhanced only through appropriately adjusting the dilation, even in the case of very low SNR. Therefore, CWT method is a powerful tool for performing the approximate derivative calculation of signals in analytical chemistry. Additionally, the approximate second derivative evaluated via CWT method can be used to determine the peak potentials of the overlapping square wave voltammogram (SWV) of Cd(II) and In(III), and the results are very satisfactory. PMID- 11911698 TI - E-state modeling of HIV-1 protease inhibitor binding independent of 3D information. AB - Data for HIV-1 protease inhibitors (in vitro enzyme binding) were used as a training set to develop a QSAR model based on topological descriptors, including two hydrogen E-state indices, along with a molecular connectivity chi and a kappa shape index. A statistically satisfactory four-variable model was obtained for the 32 compounds in the training set, r2 = 0.86, s = 0.60, and q2 = 0.79, without the use of information from 3D geometries or detailed interaction energy calculations. The model was validated through the prediction of 15 compounds in the external test set, yielding a mean absolute error, MAE, = 0.82. Structure interpretation is given for each variable to assist in the design of new compounds. Structure features emphasized in the model include hydrogen bond donating ability, nonpolar groups, skeletal branching, and molecular globularity. On the basis of these statistical criteria, this E-state model may be considered useful for prediction of pIC50 values for new HIV-1 protease inhibitors. PMID- 11911699 TI - Three-dimensional quantitative structure-property relationship (3D-QSPR) models for prediction of thermodynamic properties of polychlorinated biphenyls (PCBs): enthalpy of vaporization. AB - Three-dimensional Quantitative Structure-Property Relationship (QSPR) models have been derived using Comparative Molecular Field Analysis (CoMFA) to correlate the vaporization enthalpies of a representative set of polychlorinated biphenyls (PCBs) at 298.15 K with their CoMFA-calculated physicochemical properties. Various alignment schemes, such as inertial, as is, and atom fit, were employed in this study. The CoMFA models were also developed using different partial charge formalisms, namely, electrostatic potential (ESP) charges and Gasteiger Marsili (GM) charges. The most predictive model for vaporization enthalpy (Delta(vap)H(m)(298.15 K)), with atom fit alignment and Gasteiger-Marsili charges, yielded r2 values 0.852 (cross-validated) and 0.996 (conventional). The vaporization enthalpies of PCBs increased with the number of chlorine atoms and were found to be larger for the meta- and para-substituted isomers. This model was used to predict Delta(vap)H(m)(298.15 K) of the entire set of 209 PCB congeners. PMID- 11911697 TI - Reaction space map representation of the chlorination/dechlorination reactions of polychlorobenzenes. AB - The reaction space map (RESMAP) representation is proposed for the comprehensive description of the chlorination and dechlorination reactions of polychlorobenzenes. By using the B3LYP/6-311G energies and by assuming the chemical reactions which govern the chlorination and dechlorination processes of polychlorobenzenes, the relative energies of polychlorobenzenes and polychlorophenyl radicals were defined artificially. They were collected as the RESMAP which has the relative energies of polychlorobenzenes at the diagonal parts and the polychlorophenyl radicals connecting two polychlorobenzenes at the off-diagonal parts. The RESMAPs created for three models for the chlorination/dechlorination processes provided a general view of the thermodynamically controlled isomer distributions and chlorination/dechlorination reaction patterns for benzene and (poly)chlorinated benzenes. PMID- 11911700 TI - Heuristics for similarity searching of chemical graphs using a maximum common edge subgraph algorithm. AB - Recently a method (RASCAL) for determining graph similarity using a maximum common edge subgraph algorithm has been proposed which has proven to be very efficient when used to calculate the relative similarity of chemical structures represented as graphs. This paper describes heuristics which simplify a RASCAL similarity calculation by taking advantage of certain properties specific to chemical graph representations of molecular structure. These heuristics are shown experimentally to increase the efficiency of the algorithm, especially at more distant values of chemical graph similarity. PMID- 11911701 TI - Using molecular quantum similarity measures under stochastic transformation to describe physical properties of molecular systems. AB - The application of molecular quantum similarity measures (MQSM) to correlate physicochemical properties is reported. Satisfactory quantitative structure property relationship (QSPR) models are obtained for three molecular sets, where boiling points and chromatographic retention times and indices are studied. In this work, MQSM are scaled using a stochastic transformation and related to molecular properties using the partial least-squares technique. PMID- 11911702 TI - Analytical estimation of scaling behavior for the entanglement complexity of a bond network. AB - Geometrical entanglements in a polymer network can be characterized in terms of the mean number of projected bond-bond crossings, N. Here, we present an analytical method to study the dependence of N on the number of bonds in the network, n. Our approach shows the occurrence of power-law scaling, N - n(beta). The estimated upper bound to the exponent for maximally compact networks, Beta approximately 1.38, agrees well with the values observed in simulations of transient networks in liquids and in the folding features of native states of globular proteins. PMID- 11911703 TI - Predicting Caco-2 cell permeation coefficients of organic molecules using membrane-interaction QSAR analysis. AB - A methodology termed membrane-interaction QSAR (MI-QSAR) analysis has been developed in order to predict the behavior of organic compounds interacting with the phospholipid-rich regions of biological membranes. One important application of MI-QSAR analysis is to estimate ADME properties including the transport of organic solutes through biological membranes as a computational approach to forecasting drug intestinal absorption. A training set of 30 structurally diverse drugs, whose permeability coefficients across the cellular membranes of Caco-2 cells were measured, was used to construct significant MI-QSAR models of Caco-2 cell permeation. Cellular permeation is found to depend primarily upon aqueous solvation free energy (solubility) of the drug, the extent of drug interaction with a model phospholipid (DMPC) monolayer, and the conformational flexibility of the solute within the model membrane. A test set of eight drugs was used to evaluate the predictivity of the MI-QSAR models. The permeation coefficients of the test set compounds were predicted with the same accuracy as the compounds of the training set. PMID- 11911704 TI - An integrated SOM-fuzzy ARTMAP neural system for the evaluation of toxicity. AB - Self-organized maps (SOM) have been applied to analyze the similarities of chemical compounds and to select from a given pool of descriptors the smallest and more relevant subset needed to build robust QSAR models based on fuzzy ARTMAP. First, the category maps for each molecular descriptor and for the target activity variable were created with SOM and then classified on the basis of topology and nonlinear distribution. The best subset of descriptors was obtained by choosing from each cluster the index with the highest correlation with the target variable and then in order of decreasing correlation. This process was terminated when a dissimilarity measure increased, indicating that the inclusion of more molecular indices would not add supplementary information. The optimal subset of descriptors was used as input to a fuzzy ARTMAP architecture modified to effect predictive capabilities. The performance of the integrated SOM-fuzzy ARTMAP approach was evaluated with the prediction of the acute toxicity LC50 of a homogeneous set of 69 benzene derivatives in the fathead minnow and the oral rat toxicity LD50 of a heterogeneous set of 155 organic compounds. The proposed methodology minimized the problem of misclassification of similar compounds and significantly enhanced the predictive capabilities of a properly trained fuzzy ARTMAP network. PMID- 11911705 TI - A general QSPR treatment for dielectric constants of organic compounds. AB - Multilinear regression and neural network methods have been used to develop QSPR models for the prediction of the dielectric constant (epsilon) and Kirkwood function (epsilon - 1)/(2epsilon + 1) of organic liquids. Both methods can provide acceptable models for the prediction of these properties. The QSPR models developed from the training set of 155 diverse compounds use theoretical molecular descriptors encoding electronic properties of the molecule and the intermolecular interaction between molecules. The QSPR models for the Kirkwood function appear to be more reliable than the models for the dielectric constant. The average prediction error of the best model for the dielectric constant is 27.0%. The average prediction error of the best model for the Kirkwood function is 4.1%. PMID- 11911706 TI - The estimation of melting points and fusion enthalpies using experimental solubilities, estimated total phase change entropies, and mobile order and disorder theory. AB - Melting points and fusion enthalpies are predicted for a series of 81 compounds by combining experimental solubilities in a variety of solvents and analyzed according to the theory of mobile order and disorder (MOD) and using the total phase change entropy estimated by a group additivity method. The error associated in predicting melting points is dependent on the magnitude of the temperature predicted. An error of +/- 12 K (+/- 1 sigma) was obtained for compounds melting between ambient temperature and 350 K (24 entries). This error increased to +/- 23 K when the temperature range was expanded to 400 K (46 entries) and +/- 39 K for the temperature range 298-555 K (79 entries). Fusion enthalpies were predicted within +/- 2sigma of the experimental values (+/- 6.4 kJ mol(-1)) for 79 entries. The uncertainty in the fusion enthalpy did not appear dependent on the magnitude of the melting point. Two outliers, adamantane and camphor, have significant phase transitions that occur below room temperature. Estimates of melting temperature and fusion enthalpy for these compounds were characterized by significantly larger errors. PMID- 11911707 TI - Combinatorial library design using a multiobjective genetic algorithm. AB - Early results from screening combinatorial libraries have been disappointing with libraries either failing to deliver the improved hit rates that were expected or resulting in hits with characteristics that make them undesirable as lead compounds. Consequently, the focus in library design has shifted toward designing libraries that are optimized on multiple properties simultaneously, for example, diversity and "druglike" physicochemical properties. Here we describe the program MoSELECT that is based on a multiobjective genetic algorithm and which is able to suggest a family of solutions to multiobjective library design where all the solutions are equally valid and each represents a different compromise between the objectives. MoSELECT also allows the relationships between the different objectives to be explored with competing objectives easily identified. The library designer can then make an informed choice on which solution(s) to explore. Various performance characteristics of MoSELECT are reported based on a number of different combinatorial libraries. PMID- 11911709 TI - On combining recursive partitioning and simulated annealing to detect groups of biologically active compounds. AB - Statistical data mining methods have proven to be powerful tools for investigating correlations between molecular structure and biological activity. Recursive partitioning (RP), in particular, offers several advantages in mining large, diverse data sets resulting from high throughput screening. When used with binary molecular descriptors, the standard implementation of RP splits on single descriptors. We use simulated annealing (SA) to find combinations of molecular descriptors whose simultaneous presence best separates off the most active, chemically similar group of compounds. The search is incorporated into a recursive partitioning design to produce a regression tree for biological activity on the space of structural fingerprints. Each node is characterized by a specific combination of structural features, and the terminal nodes with high average activities correspond, roughly, to different classes of compounds. Using LeadScope structural features as descriptors to mine a database from the National Cancer Institute, the merging of RP and SA consistently identifies structurally homogeneous classes of highly potent anticancer agents. PMID- 11911708 TI - Electronic structure of some adenosine receptor antagonists. VQSAR Investigation. AB - A QSAR model has been developed for 1,3-dimethylxanthines as adenosine receptor antagonists. The model is capable of predicting the affinity toward both the A1 and A2 receptors. Constitutional, geometrical, topological, electronic descriptors (computed at the ab initio 6-31G level), and some empirical descriptors related to the hypophilicity were computed and analyzed. A two step computational strategy was adopted to select the descriptors relevant to the A1 or the A2 affinity. In the first step, each of the four main groups of descriptors is treated independently. Multiple regression analysis lead to a set of equations that reflect the weight of each of the studied descriptors. The most relevant of these descriptors were grouped, and a new multiple regression analysis has been carried out and arrived at the final QSAR model. These QSAR equations account for almost all the A2 and an appreciable part of the A1 affinity. The proposed model has been examined as a general tool of predicting the activity toward the adenosine receptor sites. A validation set of 22 xanthines were selected, and their activities were computed using the proposed QSAR model. The correspondence between the predicted and observed activities is excellent. Anova statistical analysis on the data of the validation set elaborates on the quality of these fits. PMID- 11911710 TI - A versatile structural domain analysis server using profile weight matrices. AB - The WEB tool "AnDom" assigns to a given protein sequence all experimentally determined structural domains contained within it, including multidomain and large proteins. The server uses profile specific matrices from custom generated multiple sequence alignments of all known SCOP domains (SCOP version 1.50). Prediction time is short allowing numerous applications for structural genomics including investigation of complex eucaryotic protein families. The WWW server is at http://www.bork.embl-heidelberg.de/AnDom, and profiles can be downloaded at ftp.bork.embl-heidelberg.de/pub/users/ schmidt/AnDom. PMID- 11911711 TI - The "latent membrane permeability" concept: QSPR analysis of inter/intralaboratory variable Caco-2 permeability. AB - Caco-2 cell monolayers grown on a filter support are the most widely used systems for predicting intestinal absorption. However, inter- and intralaboratory variability in Caco-2 permeability makes it difficult to analyze any QSPR relationship for a large number of compounds. We proposed the "latent membrane permeability" concept, assuming that all Caco-2 permeability data sets share a hidden, common relationship between their membrane permeability and physicochemical properties. An iterative calculation method was developed to handle this conceptual approach and applied to the analysis of Caco-2 permeability data sets from different sources. A thorough statistical analysis revealed that the "latent membrane permeability" concept be reasonable. PMID- 11911712 TI - Chemical library subset selection algorithms: a unified derivation using spatial statistics. AB - If similar compounds have similar activity, rational subset selection becomes superior to random selection in screening for pharmacological lead discovery programs. Traditional approaches to this experimental design problem fall into two classes: (i) a linear or quadratic response function is assumed (ii) some space filling criterion is optimized. The assumptions underlying the first approach are clear but not always defendable; the second approach yields more intuitive designs but lacks a clear theoretical foundation. We model activity in a bioassay as realization of a stochastic process and use the best linear unbiased estimator to construct spatial sampling designs that optimize the integrated mean square prediction error, the maximum mean square prediction error, or the entropy. We argue that our approach constitutes a unifying framework encompassing most proposed techniques as limiting cases and sheds light on their underlying assumptions. In particular, vector quantization is obtained, in dimensions up to eight, in the limiting case of very smooth response surfaces for the integrated mean square error criterion. Closest packing is obtained for very rough surfaces under the integrated mean square error and entropy criteria. We suggest to use either the integrated mean square prediction error or the entropy as optimization criteria rather than approximations thereof and propose a scheme for direct iterative minimization of the integrated mean square prediction error. Finally, we discuss how the quality of chemical descriptors manifests itself and clarify the assumptions underlying the selection of diverse or representative subsets. PMID- 11911713 TI - Molecular design based on 3D-pharmacophore. Application to 5-HT subtypes receptors. AB - A first definition of a pharmacophore for the serotonin reuptake inhibitors was carried out by considering a three-dimensional model which correlates the chemical structures of series of reuptake inhibitors with their biological affinities. A molecular design was described by analyzing two different 3D serotonin pharmacophores. This successful approach enabled us to consider the design of new serotonin ligands by the same method. PMID- 11911714 TI - METAPRINT: a metabolic fingerprint. Application to cassette design for high throughput ADME screening. AB - METAPRINT, a metabolic fingerprint, has been developed by predicting metabolic pathways and corresponding potential metabolites. Calculated drug-likeness parameters (log P and MW) have been incorporated into METAPRINT to allow the encoding of metabolic diversity within a chemical library. The application of METAPRINT in the design of cassette dosing experiments is demonstrated using a library of alpha-1a antagonists synthesized at Glaxo Wellcome. Results obtained by Ward's clustering algorithm suggest that METAPRINTs are able to discriminate between low- and high-clearance compounds. Cassette design was performed by maximizing the intracassette Euclidean distances between compounds in METAPRINT space, using simulated annealing. Calculated distances in METAPRINT space were in accordance with experimental data. PMID- 11911715 TI - Conflicts of interest: science, money, and health. PMID- 11911716 TI - Nasal myiasis in an intensive care unit linked to hospital-wide mouse infestation. AB - A large city hospital experienced an infestation of mice combated in part by broadcasting poisoned baits. Months later there was an invasion of flies into the hospital, and 2 comatose patients in an intensive care unit contracted nasal maggots. Adult flies were trapped and maggots removed from the nares of the second patient. These were identified as the green blowfly (Phaenicia sericata). Recent downsizing of hospital personnel had led to the unintended and unrecognized loss of housekeeping services in the canteen food storage areas. A mouse infestation of the hospital occurred, with the epicenter in the canteen area. This was initially addressed by scattering poisoned bait and using rodent glue boards. The result of such treatment was the presence of numerous mouse carcasses scattered throughout the building attracting the green blowfly. Adult gravid female flies trapped in the new intensive care unit (where mice were not present) laid eggs in the fetid nasal discharge of 2 comatose patients. Live trapping of mice and removal of carcasses led to an abatement of the fly infestation. The cause-and-effect nature of the mouse carcasses and flies was underscored a year later when an outbreak of P. sericata occurred in the operating department and was linked to the presence of mouse carcasses on glue boards not removed the previous fall. Hence, the disruption or loss of 1 vital link in hospital organization (in this case, housekeeping support) may lead to an unintended and bizarre outcome. PMID- 11911717 TI - Benefits of beta-blocker therapy for heart failure: weighing the evidence. AB - Our understanding of factors contributing to the progression of heart failure has advanced dramatically over the past 2 decades. We have also gained considerable insight into the pharmacology of beta-adrenergic receptor blockers (beta blockers). Based on this knowledge, we can now appreciate the potential of these drugs for the treatment of heart failure. Several beta-blockers have been shown to be clinically effective in the treatment of heart failure. Critical evaluation of the evidence from basic research studies, as well as clinical trials in patients with heart failure, helps to delineate the theoretical and clinical benefits of beta-blockers. PMID- 11911718 TI - Worklife and satisfaction of general internists. AB - BACKGROUND: Prior studies have reported relatively low job satisfaction for general internists. We used data from a large US physician survey to assess correlates of satisfaction of general internists. METHODS: The Physician Worklife Survey was mailed to a national random stratified sample of 5704 US physicians. General internists were assessed for their satisfaction, training, patient mix, work hours, the likelihood of recommending their specialty to medical students, and job stability. We then compared them with a specialist sample (internal medicine subspecialists [IMSSs]) and a primary care sample (family physicians [FPs]). Logistic regression was used to model predictors of satisfaction, stress, and medical student recruitment. RESULTS: There were 2326 respondents (adjusted response rate, 52%): 450 (19%) were general internists; 502 (22%), FPs; and 438 (19%), IMSSs. General internists were less satisfied than were IMSSs with their relationships with colleagues and with patient care issues (P<.01 for both) and less satisfied than were FPs with community ties (P =.001). Global job, career, and specialty satisfaction were significantly lower for general internists vs FPs and IMSSs (P<.05). General internists spent proportionately more of their work week in the hospital than did FPs (20% vs 13%; P<.001) and more time providing outpatient care than did IMSSs (56% vs 42%; P<.001). General internists had more patients with complex medical and psychosocial problems than did FPs (P<.01) but fewer patients with complex medical problems than did IMSSs (P<.001). Higher satisfaction for general internists was associated with older physician age, less time pressure during office visits, fewer work hours, and fewer patients with complex psychosocial problems (P<.05 for all). General internists were less likely than were FPs to recommend their specialty to medical students (P<.001). Specialty satisfaction, female gender, and control of hassles predicted medical student recruitment by general internists. CONCLUSIONS: General internists' role of caring for patients with complex problems is associated with lower levels of satisfaction than for IMSSs and FPs. Adjusting caseload for patient complexity, expanding time for office visits, and additional training in the care of patients with psychosocially complex problems may improve the job satisfaction of general internists and medical student recruitment into the specialty. PMID- 11911719 TI - Coffee intake and risk of hypertension: the Johns Hopkins precursors study. AB - BACKGROUND: Whether the increase in blood pressure with coffee drinking seen in clinical trials persists over time and translates into an increased incidence of hypertension is not known. METHODS: We assessed coffee intake in a cohort of 1017 white male former medical students (mean age, 26 years) in graduating classes from 1948 to 1964 up to 11 times over a median follow-up of 33 years. Blood pressure and incidence of hypertension were determined annually by self-report, demonstrated to be accurate in this cohort. RESULTS: Consumption of 1 cup of coffee a day raised systolic blood pressure by 0.19 mm Hg (95% confidence interval, 0.02-0.35) and diastolic pressure by 0.27 mm Hg (95% confidence interval, 0.15-0.39) after adjustment for parental incidence of hypertension and time-dependent body mass index, cigarette smoking, alcohol drinking, and physical activity in analyses using generalized estimating equations. Compared with nondrinkers at baseline, coffee drinkers had a greater incidence of hypertension during follow-up (18.8% vs. 28.3%; P =.03). Relative risk (95% confidence interval) of hypertension associated with drinking 5 or more cups a day was 1.35 (0.87-2.08) for baseline intake and 1.60 (1.06-2.40) for intake over follow-up. After adjustment for the variables listed above, however, these associations were not statistically significant. CONCLUSION: Over many years of follow-up, coffee drinking is associated with small increases in blood pressure, but appears to play a small role in the development of hypertension. PMID- 11911720 TI - Bone mass response to discontinuation of long-term hormone replacement therapy: results from the Postmenopausal Estrogen/Progestin Interventions (PEPI) Safety Follow-up Study. AB - BACKGROUND: Accelerated bone loss after stopping hormone therapy (HRT) is postulated to explain the lack of hip-fracture protection conferred by former HRT use. The abbreviation HRT (traditionally standing for "hormone replacement therapy") is used herein because of its wide recognition in the field. However, the pharmacological doses of estrogens and progestins used are not truly "replacement" in nature. OBJECTIVES: To determine whether women lose bone mineral density (BMD) after stopping HRT; to assess whether their rate of loss is significantly greater than that of women not undergoing HRT; and to determine whether long-term HRT is associated with continued gains in BMD. METHODS: A total of 495 women who were adherent to assigned treatment in the 3-year Postmenopausal Estrogen/Progestin Interventions randomized controlled trial (PEPI-RCT) and who had an additional BMD measurement during the PEPI Safety Follow-up Study were observed for an average of 3 years during and 4 years after the PEPI-RCT. RESULTS: Women who stopped HRT after 1 year during the PEPI-RCT had annual rates of BMD change of -0.54% (hip) and -0.81% (spine) during the following 2 years. Those who underwent HRT for 3 years during the PEPI-RCT and then discontinued it had annual changes of -1.01% (hip) and -1.04% (spine). Rates of BMD loss among women who stopped HRT during or after the PEPI-RCT did not differ significantly from those of women who did not undergo HRT, who lost bone at a rate of approximately 1% yearly during the first year of the PEPI-RCT and about half that rate afterward. Women who continued HRT after the PEPI-RCT did not show additional BMD gains. CONCLUSIONS: Our results do not support the hypothesis that bone is lost at an unusually fast rate after discontinuation of HRT, nor do they suggest that longer-term HRT leads to additional BMD gain beyond that evident after 3 years. PMID- 11911721 TI - Improved cardiorespiratory endurance following 6 months of resistance exercise in elderly men and women. AB - OBJECTIVE: To examine the effect of 6 months of high- or low-intensity resistance exercise on aerobic capacity and treadmill time to exhaustion in adults aged 60 to 83 years. METHODS: Sixty-two men and women completed the study protocol. Subjects were matched for strength and randomly assigned to a control (n = 16), low-intensity exercise (LEX, n = 24), or high-intensity exercise (HEX, n = 22) group. Subjects trained at either 50% of their one repetition maximum (1-RM) for 13 repetitions (LEX) or 80% of 1-RM for 8 repetitions (HEX) 3 times per week for 24 weeks. One set each of 12 exercises was performed. Strength was measured for the leg press, chest press, leg curl, leg extension, overhead press, biceps curl, seated row, and triceps dip. Muscular endurance was measured for the leg press and chest press. Aerobic capacity (peak oxygen consumption [VO(2)peak]) was measured during an incremental treadmill test (Naughton). Treadmill time to exhaustion was measured as the time to exhaustion during the incremental exercise test. RESULTS: The 1-RM significantly increased (P< or =.05) for all exercises tested for both the HEX and LEX groups. Aerobic capacity increased (P< or =.05) by 23.5% (20.2 to 24.7 mL x kg(-1) x min(-1)) and by 20.1% (20.9 to 24.4 mL x kg( 1) x min(-1)) for the LEX and HEX groups, respectively. Treadmill time increased (P< or =.05) by 26.4% and 23.3% for the LEX and HEX groups, respectively. CONCLUSIONS: Significant improvements in aerobic capacity and treadmill time to exhaustion can be obtained in older adults as a consequence of either high- or low-intensity resistance exercise. These findings suggest that increased strength, as a consequence of resistance exercise training, may allow older adults to reach and/or improve their aerobic capacity. PMID- 11911722 TI - Rapid antibiotic delivery and appropriate antibiotic selection reduce length of hospital stay of patients with community-acquired pneumonia: link between quality of care and resource utilization. AB - OBJECTIVES: To measure quality-of-care variables relevant to the treatment of community-acquired pneumonia and to determine their relative contribution to variation in length of hospital stay (LOS). METHODS: One hundred cases of pneumonia requiring hospitalization from each of 7 institutions (2 community and 5 university teaching hospitals) were randomly selected (total sample, 700 cases). Demographic and clinical variables were abstracted using a standardized data instrument. Three quality-of-care measures were analyzed: (1) site of initial antibiotic treatment (emergency department vs floor), (2) door-to-needle time, and (3) appropriateness of antibiotic selection. Appropriate antibiotic selection was defined by the 1998 Infectious Disease Society of America guidelines for the treatment of hospitalized pneumonia cases. Regression modeling was used to determine associations between LOS and our quality-of-care (process) variables. RESULTS: The mean +/- SD LOS for this sample was 7.0 +/- 4.1 days. Prolonged LOS, defined as greater than or equal to the 75th percentile of the LOS distribution, was the dependent variable in our regression analysis and was greater than or equal to 9.0 days. After clinical and demographic variables were adjusted for, logistic regression modeling revealed that all 3 quality-of-care measures were associated with prolonged LOS: (1) initial antibiotic treatment in the emergency department (odds ratio [OR], 0.31; 95% confidence interval [CI], 0.19-0.48); (2) appropriate antibiotic selection (OR, 0.55; 95% CI, 0.35-0.88); and (3) door-to-needle time (OR, 1.75 per 8 hours; 95% CI, 1.34-2.29). In a secondary analysis, we examined the clinical and demographic characteristics of the patients who were treated more rapidly in the emergency department compared with those who were treated on the inpatient floor. No clinically meaningful differences were observed between these groups. CONCLUSIONS: Unlike clinical and demographic variables, process-of-care variables are modifiable and amenable to quality improvement. We observed that rapid antibiotic initiation and appropriate antibiotic selection in the emergency department have a statistically significant association with shorter LOS. These findings suggest quality improvement targeted at these processes of care may improve resource utilization and reduce LOS for patients with community-acquired pneumonia. PMID- 11911723 TI - Gastroenteritis-associated hyperamylasemia: prevalence and clinical significance. AB - BACKGROUND: Serum amylase levels can be elevated in various pathological conditions. However, acute gastroenteritis has not been widely recognized as a cause for hyperamylasemia. PATIENTS AND METHODS: We conducted a retrospective study of amylase results for all patients hospitalized or discharged from the emergency department with a diagnosis of gastroenteritis from April through November 1999. Patients with other possible medical causes for elevated amylase levels were excluded. We also compared the clinical and laboratory parameters of hyperamylasemic vs. normoamylasemic hospitalized patients with gastroenteritis. RESULTS: A total of 1041 patients with acute gastroenteritis were identified. Serum amylase levels were determined in 701 patients and were abnormally elevated in 66 of them. In 15 patients, other possible causes of hyperamylasemia were present, and these patients were excluded. The mean serum amylase level among the remaining 51 patients (7.4% of the remaining 686 patients with gastroenteritis) was 1.32 of the upper normal level, with a range of up to 2.2 times the upper normal range. Clinicians tended to admit more hyperamylasemic patients than normoamylasemic patients (10 of 51 vs. 65 of 635; P =.03, 1 sided). However, the course of gastroenteritis in the hospitalized hyperamylasemic patients did not differ significantly from that in the hospitalized normoamylasemic patients, as judged by the clinical signs and symptoms, laboratory results, length of hospital stay, and need for antibiotics. CONCLUSIONS: Gastroenteritis is associated with a mild to moderate elevation of serum amylase levels in a significant portion of patients and should be included in the differential diagnosis of hyperamylasemia. Such elevation, however, does not seem to bear clinical significance in terms of the severity of disease. PMID- 11911724 TI - Neurological involvement in acute Q fever: a report of 29 cases and review of the literature. AB - BACKGROUND: Q fever is characterized by its clinical polymorphism; neurological involvement has occasionally been described. In the course of acute Q fever, neurological manifestations may include aseptic meningitis, encephalitis or encephalomyelitis, and peripheral neuropathy. OBJECTIVE: To review and evaluate cases of acute Q fever with neurological symptoms diagnosed in our laboratory. METHODS: A total of 1269 acute Q fever cases were recorded from January 1985 to January 2000 in our laboratory and were reviewed for neurological complications. Patients were considered to have acute Q fever when serological procedures showed Coxiella burnetii phase II titers of 1:200 or higher for IgG and 1:50 or higher for IgM. Those patients who underwent a lumbar puncture for cerebrospinal fluid analysis or who had abnormal neurological symptoms were selected for this study. We describe the clinical, epidemiological, and biological features of these cases. We also review the literature and compare our cases with those previously reported. RESULTS: Among the 45 patients selected, 14 were excluded because they had normal cerebrospinal fluid and no neurological symptoms. Two were excluded because there were no clinical or epidemiological data. Three major clinical syndromes were observed: meningoencephalitis or encephalitis in 17 cases; meningitis in 8; and myelitis and peripheral neuropathy in 4. Encephalitic signs were not specific, but behavior or psychiatric disturbances were common. CONCLUSIONS: Q fever should be included in the differential diagnosis of acute neurological disease in a patient with a fever. Serological testing should be performed in cases of meningoencephalitis, lymphocytic meningitis, and peripheral neuropathy, including Guillain-Barre syndrome and myelitis. PMID- 11911725 TI - Q fever during pregnancy: diagnosis, treatment, and follow-up. AB - BACKGROUND: Q fever, caused by Coxiella burnetii, may result in abortions, premature deliveries, and stillbirths in infected pregnant women. OBJECTIVE: To evaluate the best treatment strategy for Q fever during pregnancy. METHODS: We evaluated the prognosis of 17 pregnant women who developed Q fever with and without co-trimoxazole (trimethoprim-sulfamethoxazole) treatment. RESULTS: The outcome of the pregnancy was found to depend on the trimester. Abortions occurred in 7 of 7 insufficiently treated patients infected during the first trimester vs 1 of 5 patients infected later. Co-trimoxazole given until delivery protected against abortion (0/4) but not against the development of chronic infections, and it did not significantly reduce the colonization of the placenta (2/4 vs 4/4). CONCLUSIONS: Our results show that C burnetii infections cause abortion and that women who develop Q fever while pregnant should be treated with co-trimoxazole for the duration of pregnancy, specifically when infected during the first trimester. PMID- 11911727 TI - Aseptic meningitis associated with rofecoxib. AB - Rofecoxib is a nonsteroidal anti-inflammatory drug that is reported to act by selectively inhibiting cyclooxygenase-2. A review and analysis of reports sent to the Spontaneous Reporting System of the Food and Drug Administration, Rockville, Md, suggest that aseptic meningitis is associated with rofecoxib use. To our knowledge, there have been no published reports of aseptic meningitis occurring in association with rofecoxib use to date. We report 5 serious cases of aseptic meningitis associated with rofecoxib use. PMID- 11911726 TI - Effect of a standardized nurse case-management telephone intervention on resource use in patients with chronic heart failure. AB - BACKGROUND: Case management is believed to promote continuity of care and decrease hospitalization rates, although few controlled trials have tested this approach. OBJECTIVE: To assess the effectiveness of a standardized telephonic case-management intervention in decreasing resource use in patients with chronic heart failure. METHODS: A randomized controlled clinical trial was used to assess the effect of telephonic case management on resource use. Patients were identified at hospitalization and assigned to receive 6 months of intervention (n = 130) or usual care (n = 228) based on the group to which their physician was randomized. Hospitalization rates, readmission rates, hospital days, days to first rehospitalization, multiple readmissions, emergency department visits, inpatient costs, outpatient resource use, and patient satisfaction were measured at 3 and 6 months. RESULTS: The heart failure hospitalization rate was 45.7% lower in the intervention group at 3 months (P =.03) and 47.8% lower at 6 months (P =.01). Heart failure hospital days (P =.03) and multiple readmissions (P =.03) were significantly lower in the intervention group at 6 months. Inpatient heart failure costs were 45.5% lower at 6 months (P =.04). A cost saving was realized even after intervention costs were deducted. There was no evidence of cost shifting to the outpatient setting. Patient satisfaction with care was higher in the intervention group. CONCLUSIONS: The reduction in hospitalizations, costs, and other resource use achieved using standardized telephonic case management in the early months after a heart failure admission is greater than that usually achieved with pharmaceutical therapy and comparable with other disease management approaches. PMID- 11911728 TI - A new hospital patient care model for the new millennium: preliminary Mayo Clinic experience. PMID- 11911729 TI - Osler, old age, and dock. PMID- 11911730 TI - No cutoff date for active pursuits. PMID- 11911731 TI - Does anticoagulant therapy reduce mortality of acute pulmonary embolism? PMID- 11911733 TI - Association of diphenhydramine use with adverse effects in hospitalized older patients: possible confounders. PMID- 11911736 TI - Laparoscopic advances in general surgery. PMID- 11911737 TI - New threats and old enemies: challenges for critical care medicine. PMID- 11911738 TI - Chronic disease activists target arthritis, asthma. PMID- 11911739 TI - Despite finding anthrax vaccine useful, IOM recommends seeking a better one. PMID- 11911744 TI - Priorities for lung transplantation among patients with cystic fibrosis. PMID- 11911745 TI - Priorities for lung transplantation among patients with cystic fibrosis. PMID- 11911748 TI - Relationship between Kaposi sarcoma-associated herpesvirus and HIV. PMID- 11911749 TI - Relationship between Kaposi sarcoma-associated herpesvirus and HIV. PMID- 11911750 TI - Relationship between Kaposi sarcoma-associated herpesvirus and HIV. PMID- 11911752 TI - Measurement of serum estradiol. PMID- 11911753 TI - Value of ophthalmologic examination in diagnosing temporal arteritis. PMID- 11911755 TI - Intravenous nesiritide vs nitroglycerin for treatment of decompensated congestive heart failure: a randomized controlled trial. AB - CONTEXT: Decompensated congestive heart failure (CHF) is the leading hospital discharge diagnosis in patients older than 65 years. OBJECTIVE: To compare the efficacy and safety of intravenous nesiritide, intravenous nitroglycerin, and placebo. DESIGN, SETTING, AND PATIENTS: Randomized, double-blind trial of 489 inpatients with dyspnea at rest from decompensated CHF, including 246 who received pulmonary artery catheterization, that was conducted at 55 community and academic hospitals between October 1999 and July 2000. INTERVENTIONS: Intravenous nesiritide (n = 204), intravenous nitroglycerin (n = 143), or placebo (n = 142) added to standard medications for 3 hours, followed by nesiritide (n = 278) or nitroglycerin (n = 216) added to standard medication for 24 hours. MAIN OUTCOME MEASURES: Change in pulmonary capillary wedge pressure (PCWP) among catheterized patients and patient self-evaluation of dyspnea at 3 hours after initiation of study drug among all patients. Secondary outcomes included comparisons of hemodynamic and clinical effects between nesiritide and nitroglycerin at 24 hours. RESULTS: At 3 hours, the mean (SD) decrease in PCWP from baseline was -5.8 (6.5) mm Hg for nesiritide (vs placebo, P<.001; vs nitroglycerin, P =.03), -3.8 (5.3) mm Hg for nitroglycerin (vs placebo, P =.09), and -2 (4.2) mm Hg for placebo. At 3 hours, nesiritide resulted in improvement in dyspnea compared with placebo (P =.03), but there was no significant difference in dyspnea or global clinical status with nesiritide compared with nitroglycerin. At 24 hours, the reduction in PCWP was greater in the nesiritide group (-8.2 mm Hg) than the nitroglycerin group (-6.3 mm Hg), but patients reported no significant differences in dyspnea and only modest improvement in global clinical status. CONCLUSION: When added to standard care in patients hospitalized with acutely decompensated CHF, nesiritide improves hemodynamic function and some self reported symptoms more effectively than intravenous nitroglycerin or placebo. PMID- 11911757 TI - Association between pulse pressure and mortality in patients undergoing maintenance hemodialysis. AB - CONTEXT: Although increased blood pressure is associated with adverse outcomes in the general population, elevated blood pressure is associated with decreased mortality in patients with end-stage renal disease undergoing maintenance hemodialysis. Recent investigations in the general population have demonstrated the predictive utility of pulse pressure (systolic minus diastolic blood pressure), a measure reflecting the pulsatile nature of the cardiac cycle. OBJECTIVES: To estimate the relationship between pulse pressure and mortality in patients undergoing maintenance hemodialysis and to test our hypothesis that an increasing pulse pressure would be associated with increased risk of death up to 1 year despite the inverse relationship between conventional blood pressure measures and mortality in patients with end-stage renal disease. DESIGN, SETTING, AND PATIENTS: Retrospective cohort investigation of patients with end-stage renal disease undergoing maintenance hemodialysis at 782 hemodialysis facilities throughout the United States. Of 44 069 eligible patients as of January 1, 1998, 37 069 with complete demographic data were included in the analyses of clinical and laboratory data collected from October 1 through December 31, 1997. Patients were followed up through December 31, 1998. MAIN OUTCOME MEASURES: The primary study outcome was death at 1 year. A secondary outcome was the magnitude of the pulse pressure. RESULTS: The final patient cohort was similar to national averages with respect to age, sex, race, and diabetic status. Mean (SD) pulse pressures before dialysis were 75.0 (15.0) mm Hg and 66.9 (13.9) mm Hg after dialysis. By the end of the 1-year follow-up, 5731 patients (18.4%) died. After adjusting for level of systolic blood pressure, multivariable Cox proportional hazards modeling showed a direct and consistent relationship between increasing pulse pressure and increasing death risk. Each incremental elevation of 10 mm Hg in postdialysis pulse pressure was associated with a 12% increase in the hazard for death (hazard ratio, 1.12; 95% confidence interval, 1.06-1.18). Postdialysis systolic blood pressure was inversely related to mortality with a 13% decreased hazard for death for each incremental elevation of 10 mm Hg (hazard ratio, 0.87; 95% confidence interval, 0.84-0.90). In a multivariable linear regression model, important variables directly associated with elevated pulse pressure included age, diabetes, white race, female sex, and number of years receiving dialysis (all P<.001). CONCLUSIONS: Pulse pressure is associated with risk of death in a large, nationally representative sample of patients undergoing maintenance hemodialysis. The recognition of pulse pressure as an important correlate of mortality in patients receiving dialysis highlights the need to investigate the relationship between potential therapeutic implications of conduit vessel function and clinical outcomes in patients with end-stage renal disease. PMID- 11911756 TI - Short-term intravenous milrinone for acute exacerbation of chronic heart failure: a randomized controlled trial. AB - CONTEXT: Little randomized evidence is available to guide the in-hospital management of patients with an acute exacerbation of chronic heart failure. Although intravenous inotropic therapy usually produces beneficial hemodynamic effects and is labeled for use in the care of such patients, the effect of such therapy on intermediate-term clinical outcomes is uncertain. OBJECTIVE: To prospectively test whether a strategy that includes short-term use of milrinone in addition to standard therapy can improve clinical outcomes of patients hospitalized with an exacerbation of chronic heart failure. DESIGN: Prospective, randomized, double-blind, placebo-controlled trial conducted from July 1997 through November 1999. SETTING: Seventy-eight community and tertiary care hospitals in the United States. PARTICIPANTS: A total of 951 patients admitted with an exacerbation of systolic heart failure not requiring intravenous inotropic support (mean age, 65 years; 92% with baseline New York Heart Association class III or IV; mean left ventricular ejection fraction, 23%). INTERVENTION: Patients were randomly assigned to receive a 48-hour infusion of either milrinone, 0.5 microg/kg per minute initially (n = 477), or saline placebo (n = 472). MAIN OUTCOME MEASURE: Cumulative days of hospitalization for cardiovascular cause within 60 days following randomization. RESULTS: The median number of days hospitalized for cardiovascular causes within 60 days after randomization did not differ significantly between patients given milrinone (6 days) compared with placebo (7 days; P =.71). Sustained hypotension requiring intervention (10.7% vs 3.2%; P<.001) and new atrial arrhythmias (4.6% vs 1.5%; P =.004) occurred more frequently in patients who received milrinone. The milrinone and placebo groups did not differ significantly in in-hospital mortality (3.8% vs 2.3%; P =.19), 60-day mortality (10.3% vs 8.9%; P =.41), or the composite incidence of death or readmission (35.0% vs 35.3%; P =.92) CONCLUSION: These results do not support the routine use of intravenous milrinone as an adjunct to standard therapy in the treatment of patients hospitalized for an exacerbation of chronic heart failure. PMID- 11911758 TI - Relationship between prepregnancy anthrax vaccination and pregnancy and birth outcomes among US Army women. AB - CONTEXT: Substantial concern surrounds the potential health effects of the anthrax vaccine, particularly the potential adverse effects on reproductive processes. OBJECTIVE: To determine whether receipt of anthrax vaccination by reproductive-aged women has an effect on pregnancy rates. DESIGN, SETTING, AND PATIENTS: Cohort study, based on information from a computer database, of women aged 17 to 44 years who were stationed at Fort Stewart, Ga, or Hunter Army Airfield, Ga, from January 1999 through March 2000. MAIN OUTCOME MEASURES: Pregnancy and birth rates and adverse birth outcomes. RESULTS: Of a total of 4092 women, 3136 received at least 1 dose of the anthrax vaccine. There was a total of 513 pregnancies, with 385 following at least 1 dose of anthrax vaccine. The pregnancy rate ratio (before and after adjustment for marital status, race, and age) comparing vaccinated with unvaccinated women was 0.94 (95% confidence interval [CI], 0.8-1.2; P =.60). There were 353 live births and 25 pregnancies lost to follow-up. The birth odds ratio after anthrax vaccination (before and after adjustment for marital status and age) was 0.9 (95% CI, 0.5-1.4; P =.55). After adjusting for age, the odds ratio for adverse birth outcome after receiving at least 1 dose of anthrax vaccination was 0.9 (95% CI, 0.4-2.4; P =.88). However, this study did not have sufficient power to detect adverse birth outcomes. CONCLUSION: Anthrax vaccination had no effect on pregnancy and birth rates or adverse birth outcomes. PMID- 11911759 TI - Relationship of prenatal diagnosis and pregnancy termination to overall infant mortality in Canada. AB - CONTEXT: Prenatal diagnosis and termination of affected pregnancies can prevent infant deaths due to congenital anomalies, but an effect at the population level has not been shown. OBJECTIVE: To examine the impact of recent changes in congenital anomaly-related fetal and infant deaths on overall population-based infant mortality. DESIGN, SETTING, AND SUBJECTS: Birth cohort-based study of all live births, stillbirths, and infant deaths in Canada (excluding Ontario) for 1991-1998. MAIN OUTCOME MEASURES: Cause-specific infant mortality rates and gestational age-specific fetal death rates. RESULTS: The birth cohort-based infant mortality rate fluctuated between 6.4 and 6.1 per 1000 live births between 1991 and 1995, then dropped to 5.4 per 1000 in 1996 and 5.5 per 1000 in 1997. The rate of infant death from congenital anomalies was stable between 1991 and 1995 but declined by 21% (95% confidence interval, 19%-32%) from 1.86 per 1000 in 1995 to 1.47 per 1000 in 1996 and 1997. Fetal deaths due to pregnancy termination at 20 to 23 weeks' gestation increased dramatically in 1994, while fetal deaths due to congenital anomalies at 20 to 21 weeks increased in 1995 and subsequently. Provinces/territories with high rates of fetal death due to pregnancy termination/congenital anomalies at 20 to 23 weeks had fewer infant deaths due to congenital anomalies. CONCLUSION: A large decrease in infant deaths due to congenital anomalies was associated with the most recent decline in infant mortality in Canada, suggesting that increases in prenatal diagnosis and pregnancy termination for congenital anomalies are related to decreases in overall infant mortality at the population level. PMID- 11911760 TI - A 75-year-old man with depression. PMID- 11911761 TI - A 58-year-old woman dissatisfied with her care, 2 years later. PMID- 11911762 TI - Treatment of acute heart failure: out with the old, in with the new. PMID- 11911767 TI - Factors associated with home death for individuals who receive home support services: a retrospective cohort study. AB - OBJECTIVES: To determine the factors associated with a home death among older adults who received palliative care nursing home services in the home. METHODS: The participants in this retrospective cohort study were 151 family caregivers of patients who had died approximately 9 months prior to the study telephone interview. The interview focused on the last year of life and covered two main areas, patient characteristics and informal caregiver characteristics. RESULTS: Odds ratios [OR] and 95% confidence intervals [95% CI] were used to determine which of the 15 potential informal caregiver and seven patient predictor variables were associated with dying at home. Multivariate analysis revealed that the odds of dying at home were greater when the patient lived with a caregiver [OR = 7.85; 95% CI = (2.35, 26.27)], the patient stated a preference to die at home [OR= 6.51; 95% CI = (2.66,15.95)], and the family physician made home visits [OR = 4.79; 95% CI = (1.97,11.64)]. However the odds were lower for patients who had caregivers with fair to poor health status [OR = 0.22; 95% CI = (0.07, 0.65)] and for patients who used hospital palliative care beds [OR = 0.31; 95% CI = (0.12, 0.80)]. DISCUSSION: The findings suggest that individuals who indicated a preference to die at home and resided with a healthy informal caregiver had better odds of dying at home. Home visits by a family physician were also associated with dying at home. PMID- 11911768 TI - Elastomeric proteins: structures, biomechanical properties and biological roles. Papers of a discussion meeting. 16-17 May 2001. PMID- 11911769 TI - Elastic proteins: biological roles and mechanical properties. AB - The term 'elastic protein' applies to many structural proteins with diverse functions and mechanical properties so there is room for confusion about its meaning. Elastic implies the property of elasticity, or the ability to deform reversibly without loss of energy; so elastic proteins should have high resilience. Another meaning for elastic is 'stretchy', or the ability to be deformed to large strains with little force. Thus, elastic proteins should have low stiffness. The combination of high resilience, large strains and low stiffness is characteristic of rubber-like proteins (e.g. resilin and elastin) that function in the storage of elastic-strain energy. Other elastic proteins play very different roles and have very different properties. Collagen fibres provide exceptional energy storage capacity but are not very stretchy. Mussel byssus threads and spider dragline silks are also elastic proteins because, in spite of their considerable strength and stiffness, they are remarkably stretchy. The combination of strength and extensibility, together with low resilience, gives these materials an impressive resistance to fracture (i.e. toughness), a property that allows mussels to survive crashing waves and spiders to build exquisite aerial filters. Given this range of properties and functions, it is probable that elastic proteins will provide a wealth of chemical structures and elastic mechanisms that can be exploited in novel structural materials through biotechnology. PMID- 11911770 TI - The structure and properties of gluten: an elastic protein from wheat grain. AB - The wheat gluten proteins correspond to the major storage proteins that are deposited in the starchy endosperm cells of the developing grain. These form a continuous proteinaceous matrix in the cells of the mature dry grain and are brought together to form a continuous viscoelastic network when flour is mixed with water to form dough. These viscoelastic properties underpin the utilization of wheat to give bread and other processed foods. One group of gluten proteins, the HMM subunits of glutenin, is particularly important in conferring high levels of elasticity (i.e. dough strength). These proteins are present in HMM polymers that are stabilized by disulphide bonds and are considered to form the 'elastic backbone' of gluten. However, the glutamine-rich repetitive sequences that comprise the central parts of the HMM subunits also form extensive arrays of interchain hydrogen bonds that may contribute to the elastic properties via a 'loop and train' mechanism. Genetic engineering can be used to manipulate the amount and composition of the HMM subunits, leading to either increased dough strength or to more drastic changes in gluten structure and properties. PMID- 11911771 TI - Elastomeric gradients: a hedge against stress concentration in marine holdfasts? AB - The byssal threads of marine mussels are elastomeric fibres with a great capacity for absorbing and dissipating energy. Up to 70% of the total absorbed energy can be dissipated in the byssus. Because byssal threads attach the mussel to hard inert surfaces in its habitat, they must combine the need to be good shock absorbers with appropriate matching of Young's modulus between living tissue and a hard sub-stratum such as stone - stiffnesses that can differ by five orders of magnitude. Recent data suggest that improved modulus matching and decreased stress concentration between different portions of the byssus is achieved by the use of protein gradients. Protein gradients in byssal threads are constructed using natural macromolecular chimeras having a central collagenous domain, variable flanking modules and histidine-rich amino and carboxy termini. Stiff silk-like flanking modules prevail distally, while at the animal end, rubbery modules resembling elastin predominate. In between the two thread ends there is a mix of both module types. The histidine-rich termini provide metal binding/cross linking sites, while collagen domains may confer self-assembly on all parts of the structure. A graded axial distribution of flanking modules is expected to moderate stress concentration in joined materials having disparate moduli. PMID- 11911772 TI - Biological liquid crystal elastomers. AB - Liquid crystal elastomers (LCEs) have recently been described as a new class of matter. Here we review the evidence for the novel conclusion that the fibrillar collagens and the dragline silks of orb web spiders belong to this remarkable class of materials. Unlike conventional rubbers, LCEs are ordered, rather than disordered, at rest. The identification of these biopolymers as LCEs may have a predictive value. It may explain how collagens and spider dragline silks are assembled. It may provide a detailed explanation for their mechanical properties, accounting for the variation between different members of the collagen family and between the draglines in different spider species. It may provide a basis for the design of biomimetic collagen and dragline silk analogues by genetic engineering, peptide- or classical polymer synthesis. Biological LCEs may exhibit a range of exotic properties already identified in other members of this remarkable class of materials. In this paper, the possibility that other transversely banded fibrillar proteins are also LCEs is discussed. PMID- 11911773 TI - Silk: molecular organization and control of assembly. AB - The interface between the science and engineering of biology and materials is an area of growing interest. One of the goals of this field is to utilize biological synthesis and processing of polymers as a route to gain insight into topics such as molecular recognition, self-assembly and the formation of materials with well defined architectures. The biological processes involved in polymer synthesis and assembly can offer important information on fundamental interactions involved in the formation of complex material architectures, as well as practical knowledge into new and important materials related to biomaterial uses and tissue engineering needs. Classic approaches in biology, including genetic engineering, controlled microbial physiology and enzymatic synthesis, are prototypical methods used to control polymer structure and chemistry, including stereoselectivity and regioselectivity, to degrees unattainable using traditional synthetic chemistry. This type of control can lead to detailed and systematic studies of the formation of the structural hierarchy in materials and the subsequent biological responses to these materials. PMID- 11911774 TI - Elastin: a representative ideal protein elastomer. AB - During the last half century, identification of an ideal (predominantly entropic) protein elastomer was generally thought to require that the ideal protein elastomer be a random chain network. Here, we report two new sets of data and review previous data. The first set of new data utilizes atomic force microscopy to report single-chain force-extension curves for (GVGVP)(251) and (GVGIP)(260), and provides evidence for single-chain ideal elasticity. The second class of new data provides a direct contrast between low-frequency sound absorption (0.1-10 kHz) exhibited by random-chain network elastomers and by elastin protein-based polymers. Earlier composition, dielectric relaxation (1-1000 MHz), thermoelasticity, molecular mechanics and dynamics calculations and thermodynamic and statistical mechanical analyses are presented, that combine with the new data to contrast with random-chain network rubbers and to detail the presence of regular non-random structural elements of the elastin-based systems that lose entropic elastomeric force upon thermal denaturation. The data and analyses affirm an earlier contrary argument that components of elastin, the elastic protein of the mammalian elastic fibre, and purified elastin fibre itself contain dynamic, non-random, regularly repeating structures that exhibit dominantly entropic elasticity by means of a damping of internal chain dynamics on extension. PMID- 11911775 TI - Elastin as a self-organizing biomaterial: use of recombinantly expressed human elastin polypeptides as a model for investigations of structure and self-assembly of elastin. AB - Elastin is the major extracellular matrix protein of large arteries such as the aorta, imparting characteristics of extensibility and elastic recoil. Once laid down in tissues, polymeric elastin is not subject to turnover, but is able to sustain its mechanical resilience through thousands of millions of cycles of extension and recoil. Elastin consists of ca. 36 domains with alternating hydrophobic and cross-linking characteristics. It has been suggested that these hydrophobic domains, predominantly containing glycine, proline, leucine and valine, often occurring in tandemly repeated sequences, are responsible for the ability of elastin to align monomeric chains for covalent cross-linking. We have shown that small, recombinantly expressed polypeptides based on sequences of human elastin contain sufficient information to self-organize into fibrillar structures and promote the formation of lysine-derived cross-links. These cross linked polypeptides can also be fabricated into membrane structures that have solubility and mechanical properties reminiscent of native insoluble elastin. Understanding the basis of the self-organizational ability of elastin-based polypeptides may provide important clues for the general design of self assembling biomaterials. PMID- 11911776 TI - Viscoelastic properties of collagen: synchrotron radiation investigations and structural model. AB - Collagen type I is the most abundant structural protein in tendon, skin and bone, and largely determines the mechanical behaviour of these connective tissues. To obtain a better understanding of the relationship between structure and mechanical properties, tensile tests and synchrotron X-ray scattering have been carried out simultaneously, correlating the mechanical behaviour with changes in the microstructure. Because intermolecular cross-links are thought to have a great influence on the mechanical behaviour of collagen, we also carried out experiments using cross-link-deficient tail-tendon collagen from rats fed with beta-APN, in addition to normal controls. The load-elongation curve of tendon collagen has a characteristic shape with, initially, an increasing slope, corresponding to an increasing stiffness, followed by yielding and then fracture. Cross-link-deficient collagen produces a quite different curve with a marked plateau appearing in some cases, where the length of the tendon increases at constant stress. With the use of in situ X-ray diffraction, it was possible to measure simultaneously the elongation of the collagen fibrils inside the tendon and of the tendon as a whole. The overall strain of the tendon was always larger than the strain in the individual fibrils, which demonstrates that some deformation is taking place in the matrix between fibrils. Moreover, the ratio of fibril strain to tendon strain was dependent on the applied strain rate. When the speed of deformation was increased, this ratio increased in normal collagen but generally decreased in cross-link-deficient collagen, correlating to the appearance of a plateau in the force-elongation curve indicating creep. We proposed a simple structural model, which describes the tendon at a hierarchical level, where fibrils and interfibrillar matrix act as coupled viscoelastic systems. All qualitative features of the strain-rate dependence of both normal and cross-link-deficient collagen can be reproduced within this model. This complements earlier models that considered the next smallest level of hierarchy, describing the deformation of collagen fibrils in terms of changes in their molecular packing. PMID- 11911777 TI - Role of titin in vertebrate striated muscle. AB - Titin is a giant muscle protein with a molecular weight in the megaDalton range and a contour length of more than 1 microm. Its size and location within the sarcomere structure determine its important role in the mechanism of muscle elasticity. According to the current consensus, elasticity stems directly from more than one type of spring-like behaviour of the I-band portion of the molecule. Starting from slack length, extension of the sarcomere first causes straightening of the molecule. Further extension then induces local unfolding of a unique sequence, the PEVK region, which is named due to the preponderance of these amino-acid residues. High speeds of extension and/or high forces are likely to lead to unfolding of the beta-sandwich domains from which the molecule is mainly constructed. A release of tension leads to refolding and recoiling of the polypeptide. Here, we review the literature and present new experimental material related to the role of titin in muscle elasticity. In particular, we analyse the possible influence of the arrangement and environment of titin within the sarcomere structure on its extensible behaviour. We suggest that, due to the limited conformational space, elongation and compression of the molecule within the sarcomere occur in a more ordered way or with higher viscosity and higher forces than are observed in solution studies of the isolated protein. PMID- 11911779 TI - Spinning an elastic ribbon of spider silk. AB - The Sicarid spider Loxosceles laeta spins broad but very thin ribbons of elastic silk that it uses to form a retreat and to capture prey. A structural investigation into this spider's silk and spinning apparatus shows that these ribbons are spun from a gland homologous to the major ampullate gland of orb web spiders. The Loxosceles gland is constructed from the same basic parts (separate transverse zones in the gland, a duct and spigot) as other spider silk glands but construction details are highly specialized. These differences are thought to relate to different ways of spinning silk in the two groups of spiders. Loxosceles uses conventional die extrusion, feeding a liquid dope (spinning solution) to the slit-like die to form a flat ribbon, while orb web spiders use an extrusion process in which the silk dope is processed in an elongated duct to produce a cylindrical thread. This is achieved by the combination of an initial internal draw down, well inside the duct, and a final draw down, after the silk has left the spigot. The spinning mechanism in Loxosceles may be more ancestral. PMID- 11911778 TI - Fibrillin: from microfibril assembly to biomechanical function. AB - Fibrillins form the structural framework of a unique and essential class of extracellular microfibrils that endow dynamic connective tissues with long-range elasticity. Their biological importance is emphasized by the linkage of fibrillin mutations to Marfan syndrome and related connective tissue disorders, which are associated with severe cardiovascular, ocular and skeletal defects. These microfibrils have a complex ultrastructure and it has proved a major challenge both to define their structural organization and to relate it to their biological function. However, new approaches have at last begun to reveal important insights into their molecular assembly, structural organization and biomechanical properties. This paper describes the current understanding of the molecular assembly of fibrillin molecules, the alignment of fibrillin molecules within microfibrils and the unique elastomeric properties of microfibrils. PMID- 11911781 TI - Fitness landscapes and evolvability. AB - In this paper, we develop techniques based on evolvability statistics of the fitness landscape surrounding sampled solutions. Averaging the measures over a sample of equal fitness solutions allows us to build up fitness evolvability portraits of the fitness landscape, which we show can be used to compare both the ruggedness and neutrality in a set of tunably rugged and tunably neutral landscapes. We further show that the techniques can be used with solution samples collected through both random sampling of the landscapes and online sampling during optimization. Finally, we apply the techniques to two real evolutionary electronics search spaces and highlight differences between the two search spaces, comparing with the time taken to find good solutions through search. PMID- 11911780 TI - Comparative structures and properties of elastic proteins. AB - Elastic proteins are characterized by being able to undergo significant deformation, without rupture, before returning to their original state when the stress is removed. The sequences of elastic proteins contain elastomeric domains, which comprise repeated sequences, which in many cases appear to form beta-turns. In addition, the majority also contain domains that form intermolecular cross links, which may be covalent or non-covalent. The mechanism of elasticity varies between the different proteins and appears to be related to the biological role of the protein. PMID- 11911782 TI - Evolutionary algorithms for the satisfiability problem. AB - Several evolutionary algorithms have been proposed for the satisfiability problem. We review the solution representations suggested in literature and choose the most promising one - the bit string representation - for further evaluation. An empirical comparison on commonly used benchmarks is presented for the most successful evolutionary algorithms and for WSAT, a prominent local search algorithm for the satisfiability problem. The key features of successful evolutionary algorithms are identified, thereby providing useful methodological guidelines for designing new heuristics. Our results indicate that evolutionary algorithms are competitive to WSAT. PMID- 11911783 TI - Network random keys - a tree representation scheme for genetic and evolutionary algorithms. AB - When using genetic and evolutionary algorithms for network design, choosing a good representation scheme for the construction of the genotype is important for algorithm performance. One of the most common representation schemes for networks is the characteristic vector representation. However, with encoding trees, and using crossover and mutation, invalid individuals occur that are either under- or over-specified. When constructing the offspring or repairing the invalid individuals that do not represent a tree, it is impossible to distinguish between the importance of the links that should be used. These problems can be overcome by transferring the concept of random keys from scheduling and ordering problems to the encoding of trees. This paper investigates the performance of a simple genetic algorithm (SGA) using network random keys (NetKeys) for the one-max tree and a real-world problem. The comparison between the network random keys and the characteristic vector encoding shows that despite the effects of stealth mutation, which favors the characteristic vector representation, selectorecombinative SGAs with NetKeys have some advantages for small and easy optimization problems. With more complex problems, SGAs with network random keys significantly outperform SGAs using characteristic vectors. This paper shows that random keys can be used for the encoding of trees, and that genetic algorithms using network random keys are able to solve complex tree problems much faster than when using the characteristic vector. Users should therefore be encouraged to use network random keys for the representation of trees. PMID- 11911785 TI - Ansatz for dynamical hierarchies. AB - Complex, robust functionalities can be generated naturally in at least two ways: by the assembly of structures and by the evolution of structures. This work is concerned with spontaneous formation of structures. We define the notion of dynamical hierarchies in natural systems and show the importance of this particular kind of organization for living systems. We then define a framework that enables us to formulate, investigate, and manipulate such dynamical hierarchies. This framework allows us to simultaneously investigate different levels of description together with their interrelationship, which is necessary to understand the nature of dynamical hierarchies. Our framework is then applied to a concrete and very simple formal, physicochemical, dynamical hierarchy involving water and monomers at level one, polymers and water at level two, and micelles (polymer aggregates) and water at level three. Formulating this system as a simple two-dimensional molecular dynamics (MD) lattice gas allows us within one dynamical system to demonstrate the successive emergence of two higher levels (three levels all together) of robust structures with associated properties. Second, we demonstrate how the framework for dynamical hierarchies can be used for realistic (predictive) physicochemical simulation of molecular self-assembly and self-organization processes. We discuss the detailed process of micellation using the three-dimensional MD lattice gas. Finally, from these examples we can infer principles about formal dynamical hierarchies. We present an ansatz for how to generate robust, higher-order emergent properties in formal dynamical systems that is based on a conjecture of a necessary minimal complexity within the fundamental interacting structures once a particular simulation framework is chosen. PMID- 11911786 TI - Is it the right ansatz? AB - This article is a response to Rasmussen et al. [Artificial Life, 7, 329-350], in which the authors suggest that, within a particular simulation "framework," there is a tight correspondence between the complexity of the primitive objects and the emergence of dynamical hierarchies. As an example they report a two-dimensional artificial chemistry that supports the spontaneous emergence of micellar structures, which they classify as third-order structures. We report in this article that essentially comparable phenomena can be produced with relatively simpler primitive objects. We also question the order classification of the micellar structures. PMID- 11911787 TI - Defense of the ansatz for dynamical hierarchies. AB - Gross and McMullin [Artificial Life, 7, 355-365] criticize the conclusions of our article on dynamical hierarchies [Artificial Life, 7, 329-353]. In this note we respond to their criticisms. After clarifying our ansatz, we argue that the simulations presented by Gross and McMullin present no evidence against the ansatz, in part because their simulations use a different simulation framework, and in part because their simulations are no less complex than ours. We also clarify why the micelles in our simulations are third-order emergent structures, and why we emphasize realism in our simulation. PMID- 11911788 TI - A macroscopic analytical model of collaboration in distributed robotic systems. AB - In this article, we present a macroscopic analytical model of collaboration in a group of reactive robots. The model consists of a series of coupled differential equations that describe the dynamics of group behavior. After presenting the general model, we analyze in detail a case study of collaboration, the stick pulling experiment, studied experimentally and in simulation by Ijspeert et al. [Autonomous Robots, 11, 149-171]. The robots' task is to pull sticks out of their holes, and it can be successfully achieved only through the collaboration of two robots. There is no explicit communication or coordination between the robots. Unlike microscopic simulations (sensor-based or using a probabilistic numerical model), in which computational time scales with the robot group size, the macroscopic model is computationally efficient, because its solutions are independent of robot group size. Analysis reproduces several qualitative conclusions of Ijspeert et al.: namely, the different dynamical regimes for different values of the ratio of robots to sticks, the existence of optimal control parameters that maximize system performance as a function of group size, and the transition from superlinear to sublinear performance as the number of robots is increased. PMID- 11911789 TI - Comparison of different genotype encodings for simulated three-dimensional agents. AB - We analyze the effect of different genetic encodings used for evolving three dimensional agents with physical morphologies. The complex phenotypes used in such systems often require nontrivial encodings. Different encodings used in Framsticks--a system for evolving three-dimensional agents--are presented. These include a low-level direct mapping and two higher-level encodings: one recurrent and one developmental. Quantitative results are presented from three simple optimization tasks (passive height, active height, and locomotion speed). The low level encoding produced solutions of lower fitness than the two higher-level encodings under similar conditions. Results from recurrent and developmental encodings had similar fitness values but displayed qualitative differences. Desirable advantages and some drawbacks of more complex encodings are established. PMID- 11911792 TI - Quality control in manufacturing oligo arrays: a combinatorial design approach. AB - The advent of the DNA microarray technology has brought with it the exciting possibility of simultaneously observing the expression levels of all genes in an organism. One such microarray technology, called "oligo arrays," manufactures short single strands of DNA (called probes) onto a glass surface using photolithography. An altered or missed step in such a manufacturing protocol can adversely affect all probes using this failed step and is in general impossible to disentangle from experimental variation when using such a defective array. The idea of designing special quality control probes to detect a failed step was first formulated by Hubbell and Pevzner (1999). We consider an alternative formulation of this problem and use a combinatorial design approach to solve it. Our results improve over prior work in guaranteeing coverage of all protocol steps and in being able to tolerate a greater number of unreliable probe intensities. PMID- 11911793 TI - The efficient computation of position-specific match scores with the fast fourier transform. AB - Historically, in computational biology the fast Fourier transform (FFT) has been used almost exclusively to count the number of exact letter matches between two biosequences. This paper presents an FFT algorithm that can compute the match score of a sequence against a position-specific scoring matrix (PSSM). Our algorithm finds the PSSM score simultaneously over all offsets of the PSSM with the sequence, although like all previous FFT algorithms, it still disallows gaps. Although our algorithm is presented in the context of global matching, it can be adapted to local matching without gaps. As a benchmark, our PSSM-modified FFT algorithm computed pairwise match scores. In timing experiments, our most efficient FFT implementation for pairwise scoring appeared to be 10 to 26 times faster than a traditional FFT implementation, with only a factor of 2 in the acceleration attributable to a previously known compression scheme. Many important algorithms for detecting biosequence similarities, e.g., gapped BLAST or PSIBLAST, have a heuristic screening phase that disallows gaps. This paper demonstrates that FFT algorithms merit reconsideration in these screening applications. PMID- 11911794 TI - Protein engineering study of protein L by simulation. AB - We examine the ability of our recently introduced minimalist protein model to reproduce experimentally measured thermodynamic and kinetic changes upon sequence mutation in the well-studied immunoglobulin-binding protein L. We have examined five different sequence mutations of protein L that are meant to mimic the same mutation type studied experimentally: two different mutations which disrupt the natural preference in the beta-hairpin #1 and beta-hairpin #2 turn regions, two different helix mutants where a surface polar residue in the alpha-helix has been mutated to a hydrophobic residue, and a final mutant to further probe the role of nonnative hydrophobic interactions in the folding process. These simulated mutations are analyzed in terms of various kinetic and thermodynamic changes with respect to wild type, but in addition we evaluate the structure-activity relationship of our model protein based on the phi-value calculated from both the kinetic and thermodynamic perspectives. We find that the simulated thermodynamic phi-values reproduce the experimental trends in the mutations studied and allow us to circumvent the difficult interpretation of the complicated kinetics of our model. Furthermore, the level of resolution of the model allows us to directly predict what experiments seek in regard to protein engineering studies of protein folding--namely the residues or portions of the polypeptide chain that contribute to the crucial step in the folding of the wild-type protein. PMID- 11911795 TI - Improved background correction for spotted DNA microarrays. AB - Most microarray scanning software for glass spotted arrays provides estimates for the intensity for the "foreground" and "background" of two channels for every spot. The common approach in further analyzing such data is to first subtract the background from the foreground for each channel and to use the ratio of these two results as the estimate of the expression level. The resulting ratios are, after possible averaging over replicates, the usual inputs for further data analysis, such as clustering. If, with this background correction procedure, the foreground intensity was smaller than the background intensity for a channel, that spot (on that array) yields no usable data. In this paper it is argued that this preprocessing leads to estimates of the expression that have a much larger variance than needed when the expression levels are low. PMID- 11911796 TI - Modeling and simulation of genetic regulatory systems: a literature review. AB - In order to understand the functioning of organisms on the molecular level, we need to know which genes are expressed, when and where in the organism, and to which extent. The regulation of gene expression is achieved through genetic regulatory systems structured by networks of interactions between DNA, RNA, proteins, and small molecules. As most genetic regulatory networks of interest involve many components connected through interlocking positive and negative feedback loops, an intuitive understanding of their dynamics is hard to obtain. As a consequence, formal methods and computer tools for the modeling and simulation of genetic regulatory networks will be indispensable. This paper reviews formalisms that have been employed in mathematical biology and bioinformatics to describe genetic regulatory systems, in particular directed graphs, Bayesian networks, Boolean networks and their generalizations, ordinary and partial differential equations, qualitative differential equations, stochastic equations, and rule-based formalisms. In addition, the paper discusses how these formalisms have been used in the simulation of the behavior of actual regulatory systems. PMID- 11911797 TI - Inference from clustering with application to gene-expression microarrays. AB - There are many algorithms to cluster sample data points based on nearness or a similarity measure. Often the implication is that points in different clusters come from different underlying classes, whereas those in the same cluster come from the same class. Stochastically, the underlying classes represent different random processes. The inference is that clusters represent a partition of the sample points according to which process they belong. This paper discusses a model-based clustering toolbox that evaluates cluster accuracy. Each random process is modeled as its mean plus independent noise, sample points are generated, the points are clustered, and the clustering error is the number of points clustered incorrectly according to the generating random processes. Various clustering algorithms are evaluated based on process variance and the key issue of the rate at which algorithmic performance improves with increasing numbers of experimental replications. The model means can be selected by hand to test the separability of expected types of biological expression patterns. Alternatively, the model can be seeded by real data to test the expected precision of that output or the extent of improvement in precision that replication could provide. In the latter case, a clustering algorithm is used to form clusters, and the model is seeded with the means and variances of these clusters. Other algorithms are then tested relative to the seeding algorithm. Results are averaged over various seeds. Output includes error tables and graphs, confusion matrices, principal-component plots, and validation measures. Five algorithms are studied in detail: K-means, fuzzy C-means, self-organizing maps, hierarchical Euclidean-distance-based and correlation-based clustering. The toolbox is applied to gene-expression clustering based on cDNA microarrays using real data. Expression profile graphics are generated and error analysis is displayed within the context of these profile graphics. A large amount of generated output is available over the web. PMID- 11911798 TI - Strong feature sets from small samples. AB - For small samples, classifier design algorithms typically suffer from overfitting. Given a set of features, a classifier must be designed and its error estimated. For small samples, an error estimator may be unbiased but, owing to a large variance, often give very optimistic estimates. This paper proposes mitigating the small-sample problem by designing classifiers from a probability distribution resulting from spreading the mass of the sample points to make classification more difficult, while maintaining sample geometry. The algorithm is parameterized by the variance of the spreading distribution. By increasing the spread, the algorithm finds gene sets whose classification accuracy remains strong relative to greater spreading of the sample. The error gives a measure of the strength of the feature set as a function of the spread. The algorithm yields feature sets that can distinguish the two classes, not only for the sample data, but for distributions spread beyond the sample data. For linear classifiers, the topic of the present paper, the classifiers are derived analytically from the model, thereby providing an enormous savings in computation time. The algorithm is applied to cancer classification via cDNA microarrays. In particular, the genes BRCA1 and BRCA2 are associated with a hereditary disposition to breast cancer, and the algorithm is used to find gene sets whose expressions can be used to classify BRCA1 and BRCA2 tumors. PMID- 11911799 TI - Interferon trials in small cell lung cancer at one institution: a comparison of results obtained before and after initiation of systematic treatment trials using IFN-alpha in combination with other modalities. AB - Chemotherapy became the primary treatment for small cell lung cancer (SCLC) in the early 1970s. The standard drug combinations were first vincristine, adriamycin, and cyclophosphamide (VAC) and then, from the early 1980s, etoposide platinum combinations. Despite a good initial objective response, however, patients usually suffer a rapid relapse. Treatment development has, therefore, focused on ways to overcome drug resistance, and on the addition of cytokines to the chemotherapeutic arsenal. Interferon (IFN) was one of the first cytokines found to have anticancer effects, and it was introduced into the combined modality regimens used to treat SCLC in the early 1980s in an attempt to overcome the problem of early relapse. The role of IFN was investigated with the aim of establishing how best to combine it with other treatments for SCLC. In this paper, we review the impact of IFN on the outcome for 714 SCLC patients who were treated in randomized IFN trials at one institution over a period of 20 years and IFN trials conducted at other institutions during the same period. The parameters we used at our institution to measure outcome tended to improve during the period when patients were being treated in our three randomized IFN trials, compared with the period when patients received only standard treatment in a nonclinical trial setting. However, the differences were not statistically significant. During this period, IFN was used as maintenance therapy, concomitantly with chemotherapy, and combined with other treatment modalities. Our experience is that IFN-alpha is most effective when administered as low-dose maintenance treatment. Other IFN trials published during the same period were small and heterogeneous. Results were inconsistent and added little new information, although it has been shown that high pretreatment levels of serum vascular endothelial growth factor (VEGF) predict a poor response to treatment and consequently a poor outcome. The recently confirmed antiangiogenic properties of IFN deserve to be investigated in studies of maintenance treatment, in combination with other biologic agents. Patient should be selected according to criteria based on pretreatment assessment of biologic markers, such as VEGF and basic fibroblast growth factor (bFGF). Our studies, all at one institution, pioneered the biologic treatment of solid tumors and developed a solid basis of knowledge for future studies of biologic agents in cancer treatment. PMID- 11911800 TI - Comparison of gene expression patterns induced by treatment of human umbilical vein endothelial cells with IFN-alpha 2b vs. IFN-beta 1a: understanding the functional relationship between distinct type I interferons that act through a common receptor. AB - We analyzed whether interferon-alpha 2b (IFN-alpha 2b) and IFN-beta 1a engage their common receptor to generate activated receptor complexes possessing distinct signaling properties. Human vascular endothelial cells (HUVEC) are 100 1000-fold more sensitive to IFN-beta 1a than to IFN-alpha 2b in in vitro assays. An nonarray-based expression profiling (GeneCalling) technology was employed to compare the patterns and levels of gene expression induced by these IFN as the broadest means by which signaling events could be measured. To distinguish subtype-related differences from dose-related effects, RNA was prepared from HUVEC treated with 50-5000 pg/ml of each IFN. The results showed that at 50 pg/ml IFN, only a subset of the genes induced by IFN-beta 1a were also induced by IFN alpha 2b and that individual genes were induced to higher levels by IFN-beta 1a. In contrast, at 5000 pg/ml, both subtypes induced essentially identical sets of genes to similar levels of expression. No genes were seen to be induced uniquely by IFN-alpha 2b but not by IFN-beta 1a. The results show that the two IFN are intrinsically capable of inducing similar gene induction responses and do not provide evidence that they generate activated receptor complexes possessing distinct signaling properties. In contrast, the two IFN generate gene induction patterns that are both qualitatively and quantitatively distinct at subsaturating and potentially physiologically more relevant concentrations. PMID- 11911801 TI - Signals involved in mycobacteria-induced CXCL-8 production by human monocytes. AB - CXC chemokine-interleukin-8 (IL-8) (CXCL-8) is a potent proinflammatory chemotactic factor that induces important immune responses for antimycobacterial defenses. However, little is known about the biochemical mechanisms by which the mycobacterial antigens upregulate the release of CXCL-8 from human monocytes. In this study, the mechanisms through which Mycobacterium bovis BCG induces CXCL-8 secretion in human monocytes were investigated. We found that M. bovis BCG induced the production of high levels of CXCL-8 by human monocytes. M. bovis induced CXCL-8 secretion was unaffected by the protein kinase C (PKC) inhibitor bisindolylmaleimide. In contrast, preincubation of the monocytes with the protein tyrosine kinase (PTK) inhibitor genistein resulted in dose-dependent suppression of mycobacteria-induced CXCL-8 secretion. These results were further supported by the fact that treatment of monocytes with herbimycin-A, another well-described inhibitor of PTK activity with a different mechanism of action, significantly diminished the effect of M. bovis on CXCL-8 secretion. In addition, the specificity of this inhibition was demonstrated by the inability of herbimycin-A to block in a significant manner IL-1 beta induction of CXCL-8. Herbimycin-A significantly blocked tyrosine phosphorylation of p59(hck) in response to M. bovis. Finally, two specific NF-kappa B inhibitors, sulfasalazine and caffeic acid phenetyl ester (CAPE), strongly inhibited the production of CXCL-8 by human monocytes infected with M. bovis. These results show intracellular signaling pathways and a transcription factor involved in the M. bovis-mediated upregulation of CXCL-8 biosynthesis and release by human monocytes. PMID- 11911802 TI - The proapoptotic 9-2 isozyme of 2-5 (A) synthetase cannot substitute for the sperm functions of the proapoptotic protein, Bax. AB - The 9-2 isozyme of 2-5 (A) synthetase has cellular proapoptotic functions that are mediated not by enzyme activity but by the Bcl-2 homology domain 3 present in its unique carboxyl-terminal region. Another proapoptotic cellular protein is Bax, whose absence in the Bax(-/-) mice causes male sterility due to abnormal sperm differentiation. In this study, we examined whether transgenic 9-2 expression can substitute for the in vivo reproductive function of Bax. To achieve this goal, a sperm-specific promoter was used to drive the expression of 9-2 in the sperm of transgenic mice. By selective cross-breeding, the transgene was transferred to Bax(-/-) mice to generate the experimental mouse line (Bax(-/ ), 9-2(+/+)). The male experimental mice were sterile, and their testes maintained the structural abnormality found in Bax(-/-) mice. Thus, the male reproduction functions of Bax could not be replaced by the 9-2 isozyme of 2-5 (A) synthetase. PMID- 11911803 TI - Neutralizing and binding antibodies to IFN-beta: relative frequency in relapsing remitting multiple sclerosis patients treated with different IFN-beta preparations. AB - The frequencies of anti-interferon-beta (IFN-beta) antibody development reported to date in patients treated with different IFN-beta preparations are not readily comparable mainly because of differences in underlying diseases and assay methods. Thus, the frequency of neutralizing antibody (NAb) and binding antibody (BAb) development was analyzed in a sample of sera derived from a homogeneous group of relapsing-remitting multiple sclerosis (RRMS) patients treated with different IFN-beta preparations. The frequency of developing NAb and BAb to IFN beta varied according to the IFN-beta given. Specifically, the NAb seroconversion frequency was significantly higher in patients treated with Betaferon, Schering AG, Berlin, Germany (31.3%) than in patients treated with both preparations of recombinant IFN-beta 1a (Rebif, Serono, Geneva, Switzerland [7.4%] or Avonex, Biogen, Cambridge, MA [6.3%]). Analysis of BAb seroconversion frequency in the same patients revealed that different IFN-beta preparations may also have different capability to induce BAb development and that BAb are produced during IFN-beta therapy at a significantly higher rate than NAb. Our main conclusion is that different human IFN-beta preparations may possess different immunogenicities, leading to varying frequency of development of antibody to IFN beta in RRMS. PMID- 11911804 TI - Direct influence of mild hypothermia on cytokine expression and release in cultures of human peripheral blood mononuclear cells. AB - Hypothermia is associated with elevated frequency of infectious complications. Dysfunction of the immune response caused by hypothermia has been demonstrated in both clinical and animal studies, but it still remains unclear to what extent immunocompetent cells are directly influenced by hypothermia. To estimate the direct influence of mild hypothermia on cytokine expression and release by human peripheral blood mononuclear cells (PBMC), primary cultures of PBMC were incubated at 34 degrees C or 32 degrees C activated by lipopolysaccharide (LPS), phytohemagglutinin (PHA), or tumor necrosis factor-alpha (TNF-alpha). The cytokine gene expression was evaluated by RT-PCR. Release of interleukin-2 (IL 2), IL-6, IL-10, and TNF-alpha was measured by ELISA. Mild hyperthermia significantly impaired IL-2 gene expression in PHA-stimulated cultures of PBMC and decreased IL-2 release in all variants of cultures. Secretion of IL-6, IL-10, and TNF-alpha was decreased in hypothermic cultures of PBMC stimulated with the T lymphocyte activator PHA. Slight suppression of IL-10 secretion was observed also in TNF-alpha-stimulated hypothermic cultures of PBMC. TNF-alpha release increased slightly in mild hypothermia control cultures. Our data demonstrate that the direct influence of hypothermia on cytokine expression and release from PBMC is not uniform. Reduction of IL-2 production might play a crucial role in the impairment of immune response in hypothermia. PMID- 11911805 TI - Serum profiles of C-C chemokines in acute myocardial infarction: possible implication in postinfarction left ventricular remodeling. AB - C-C chemokines are essential factors in the recruitment and activation of leukocytes from the circulation into inflamed tissue and may play a role in ischemia-induced myocardial injury and left ventricular remodeling after acute myocardial infarction (AMI). We investigated the kinetics of three major C-C chemokines, macrophage chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1 alpha (MIP-1 alpha), and regulated on activation normally T cell expressed and secreted (RANTES), in the sera of AMI patients and correlated the findings with the severity of the disease. Serum levels of C-C chemokines were determined in 35 AMI patients by ELISA assays serially during the first week of hospitalization and 1 month after hospital admission. Patients (n = 18) with uncomplicated AMI (Killip class I) were classified as group A, patients (n = 17) with AMI complicated by heart failure manifestations (Killip classes II and III) were classified as group B, and 15 age-matched and sex-matched volunteers were used as healthy controls. A sustained increase in serum C-C chemokines was observed in both AMI groups during the 7-day hospitalization period. Peaks of these inflammatory factors were significantly higher in group B than in group A (MCP-1, 295 +/- 11 vs. 203 +/- 9 pg/ml, p < 0.01; MIP-1 alpha, 30 +/- 1 vs. 24 +/ 2 pg/ml, p < 0.05; RANTES, 32 +/- 2 vs. 16 +/- 1 ng/ml, p < 0.01) and healthy controls (MCP-1, 125 +/- 7 pg/ml, p < 0.001; MIP-1 alpha, 14 +/- 1 pg/ml, p < 0.001; RANTES, 12 +/- 1 ng/ml, p < 0.001). In group B, significant correlations were found between the peak of MCP-1 and the peak of C-reactive protein levels (r = 0.55, p < 0.02) as well as wedge pressure (r = 0.40, p < 0.05). In the same group, the peak of MIP-1 alpha levels was also significantly correlated with the peak of serum creatine kinase-myocardial band (MB) (r = 0.51, p < 0.04) and left ventricular ejection fraction (LVEF) (r = -0.45, p < 0.05). After 1 month, AMI patients (n = 14) with severe left ventricular dysfunction (LVEF < or = 35%) exhibited significantly higher levels of C-C chemokines (all p < 0.05) than the other AMI patients (n = 21) (LVEF > 35%). A significant correlation was found between MIP-1 alpha levels and left ventricular end-diastolic diameter (r = 0.47, p < 0.03) in this patient population. In conclusion, we have detected a significant elevation of major C-C chemokines during the course of AMI, with the highest levels in patients with AMI complicated by heart failure manifestations and severe left ventricular dysfunction. The elevation of these chemotactic inflammatory factors may actively contribute to the pathophysiology of the disease and the subsequent left ventricular remodeling. PMID- 11911806 TI - Production of a monoclonal antibody that recognizes bovine stem cell factor (SCF) and its use in the detection and quantitation of native soluble bovine SCF in fetal bovine serum. AB - Stem cell factor (SCF) is a pluritropic hematopoietic cytokine that acts predominantly on the proliferation and differentiation of hematopoietic progenitor cells. SCF has long been thought to be present in fetal bovine serum (FBS) as an endogenous factor that stimulates the growth of hematopoietic progenitor cells in FBS-supplemented cultures. To detect and quantitate bovine SCF in FBS, we produced a monoclonal antibody (mAb) by immunizing mice with recombinant soluble bovine SCF protein, which was expressed in insect cells by using a baculovirus system. Using the mAb, we purified native soluble bovine SCF from FBS by immunoaffinity chromatography. Western blot analysis revealed that the purified SCF protein had a molecular weight of 33 kDa. In addition, an enzyme linked immunosorbent assay (ELISA) incorporating the mAb revealed that the levels of native soluble SCF in commercially available FBS were likely to be <100 pg/ml. These results suggest that the concentration of native soluble bovine SCF present in FBS may be insufficient to promote additive biologic effects on the growth of hematopoietic progenitor cells in FBS-supplemented cultures. PMID- 11911807 TI - Intracellular redistribution of interferon-inducible proteins Nmi and IFP 35 in apoptotic cells. AB - As a consequence of the activation of the apoptotic pathway, the subcellular localization of many proteins that modulate cell death is altered. Nmi and IFP 35 are homologous interferon (IFN)-inducible proteins that associate into a 300-400 kDa complex in the cytoplasm. Prior to treatment with IFN, subcellular fractionation reveals that the proteins are found primarily in a soluble cytoplasmic fraction, and immunofluorescence shows a diffuse cytoplasmic distribution. Treatment with IFN results in some Nmi and IFP 35 fractionating with membranes and colocalizing into speckles as detected by immunofluorescence. In IFN-treated apoptotic cells, the speckles dissociate and are replaced by a diffuse cytoplasmic distribution of Nmi and IFP 35. In Jurkat cells, dissociation of speckles could not be separated from other markers of apoptosis, including exposure of phosphatidylserine on the cell surface, caspase cleavage of D4-GDI, and nuclear condensation and fragmentation. The caspase inhibitor, zVAD-fmk, was not able to inhibit the dissociation of speckles in vivo or in a permeabilized cell assay in vitro, suggesting that dissociation of speckles is a caspase independent process. Nmi and IFP 35 proteins are not caspase substrates, as total levels of Nmi and IFP 35 and their association into a cytoplasmic complex do not change in apoptotic cells. Consistent with dissociation of speckles, subcellular fractionation studies show redistribution of Nmi and IFP 35 from the membrane fraction into the S100 fraction of apoptotic cells. Our studies have revealed dissociation of the IFN-induced Nmi/IFP 35 speckles as a newly defined specific event occurring during apoptosis. PMID- 11911808 TI - Autoantibodies in multiple sclerosis patients before and during IFN-beta 1b treatment: are they correlated with the occurrence of autoimmune diseases? AB - Autoimmune side effects, namely autoantibody (autoAb) occurrence and thyroid function alteration, have been described during interferon-beta (IFN-beta) treatment for multiple sclerosis (MS). AutoAb occurrence and autoimmune thyroid diseases are also frequently detected in MS patients free of any treatment. The aim of this study was to evaluate the relationship between IFN-beta 1b treatment, autoAb occurrence, and autoimmune diseases in MS. Thyroid and liver function and serum autoAb (antithyroid, antinuclear, anti-liver, anti-kidney microsomes, anti smooth muscle and parietal cell antigens) occurrence were evaluated in 156 relapsing-remitting MS (RRMS) patients before and every 3 months after starting IFN-beta 1b treatment (8 MIU subcutaneously [s.c.] on alternate days). The probability of having liver or thyroid function alteration or autoAb occurrence was analyzed longitudinally with the generalized estimating equations (GEE) approach. At baseline, 16.1% of patients had autoAb. During treatment, autoAb occurred de novo in 7.2% of patients. GEE analysis showed that the probability of having autoAb at any time during IFN-beta 1b treatment did not change significantly compared with baseline. AutoAb occurring de novo rarely persisted during treatment and significantly less than those already present at baseline. Positivity for autoAb at baseline or during treatment was not correlated with the development of thyroid or liver function alteration during IFN-beta 1b treatment. Our study indicates that IFN-beta treatment is a safe treatment for MS patients, free of risk of autoimmunity and of associated liver or thyroid function alteration. PMID- 11911809 TI - The CXC chemokine NAP-2 mediates differential heterologous desensitization of neutrophil effector functions elicited by platelet-activating factor. AB - During early inflammation, the chemoattractants neutrophil-activating peptide-2 (NAP-2), platelet-activating factor (PAF), and complement component C5a are rapidly generated within the vasculature and potently induce effector functions in neutrophils, such as chemotaxis and degranulation. We investigated whether these mediators would cross-desensitize each other's activities on isolated neutrophils. We demonstrate that NAP-2 and C5a desensitize degranulation of neutrophils in response to PAF. However, whereas C5a-mediated desensitization correlated with the downregulation of PAF binding sites on neutrophils, NAP-2 did not regulate PAF receptors, nor did it modulate their calcium signaling. In further contrast to C5a, which desensitized PAF-induced neutrophil chemotaxis, NAP-2 did not affect the chemotatic response to PAF. These observations indicate that NAP-2 mediates selective desensitization of PAF-induced neutrophil degranulation by a mechanism downstream from PAF receptors, still allowing migration toward PAF. Thus, a role for NAP-2 may be to limit PAF-dependent vascular tissue damage by preventing degranulation of neutrophils without affecting their migration into the inflamed tissue. PMID- 11911810 TI - Ectopic expression of Bcl-2 and Bcl-xL inhibits apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL) through suppression of caspases-8, 7, and 3 and BID cleavage in human acute myelogenous leukemia cell line HL-60. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is one of the latest members of the TNF superfamily known to induce apoptosis in a wide variety of tumor cells. Some cell types, however, are quite resistant to TRAIL. We investigated the effect of ectopic expression of Bcl-2 and Bcl-xL on TRAIL induced apoptosis in human acute myelogenous leukemia HL-60 cells. We found that HL-60 cells, which express TRAIL receptors (also called death receptor, DR) DR4, DR5, and Dc (decoy) R2, are highly sensitive to TRAIL-induced cytotoxicity. Greater than 90% killing occurred within 24 h of TRAIL treatment. The expression of Bcl-2 and Bcl-xL, however, completely abolished the TRAIL-induced cytotoxic effects. Treatment of HL-60 cells with TRAIL induced caspase-8 activation within 2-4 h, but no activation could be seen in Bcl-2-expressing or Bcl-xL-expressing cells. TRAIL also induced cleavage of BID, which was also abolished by Bcl-2 and Bcl-xL. Similarly, TRAIL activated caspase-3 and caspase-7 in control cells but not in cells expressing Bcl-2 or Bcl-xL. Cleavage of the caspase-3 substrate poly(ADP-ribose) polymerase (PARP), was abrogated by ectopic expression of Bcl-2 and Bcl-xL. Inhibition of caspases by the pan-caspase inhibitor, benzyloxycarbonyl-valine-alanine-aspartate-fluoromethylketone (zVAD-fmk) abolished the TRAIL-induced apoptosis. Overall, these results indicate that TRAIL induced apoptosis involves activation of caspase-8, caspase-7, caspase-3, and BID cleavage, and Bcl-2 and Bcl-xL prevents TRAIL-induced apoptosis by abrogating caspase activation and BID cleavage. PMID- 11911811 TI - The relationship between diabetes and periodontal disease. AB - There is good evidence to support the claim that periodontitis may be more prevalent among diabetic patients than nondiabetic people. Similarly, studies have shown that periodontal therapy influences glycemic control in people with diabetes mellitus. Given that nearly 10% of Canadians are affected by either type 1 or type 2 diabetes (including those in whom the disease is undiagnosed), all dentists will encounter patients with diabetes. Dental practitioners must be aware of the implications of this relationship and manage their patients' periodontal care accordingly. PMID- 11911812 TI - Periodontal disease and preterm delivery of low-birth-weight infants. AB - Preterm delivery of low-birth-weight infants (PLBW) remains a significant public health issue and a leading cause of neonatal death and long-term neurodevelopmental disturbances and health problems. Recent epidemiological and microbiological immunological studies have suggested that periodontal disease may be an independent risk factor for PLBW. Postulated mechanisms include translocation of periodontal pathogens to the fetoplacental unit and action of a periodontal reservoir of lipopolysaccharides or inflammatory mediators. However, non-causal explanations for the correlation between periodontitis and PLBW can also be offered. Prospective studies, and eventually interventional studies, will be necessary before periodontitis can be considered as a causal factor for PLBW. PMID- 11911813 TI - Salivary markers of systemic disease: noninvasive diagnosis of disease and monitoring of general health. AB - Because of interest in the link between oral and general health, clinicians are increasingly using salivary analyses to diagnose systemic disease and to monitor general health. The reason for this interest lies in the ability of new diagnostic tools, such as sensitive enzyme-linked immunosorbent assays, as well as other technologies, to distinguish a range of salivary components that are biomarkers for changes in the body's health. The noninvasive nature of salivary testing has made it an attractive and effective alternative to blood and urine testing, and home testing kits have made it possible for people to monitor their own health using this diagnostic medium. This paper explores what saliva can reveal about general health, drawing examples from recent research on salivary biomarkers of systemic illness and highlighting the current use, and potential clinical and research applications, of diagnostics based on oral fluids. PMID- 11911814 TI - Is periodontal disease a risk factor for coronary artery disease (CAD)? AB - Coronary artery disease (CAD) remains the principal cause of death in most developed countries, despite significant preventive and therapeutic advances. Current epidemiological data imply that recent reductions in the prevalence of this disease are unlikely to be sustained until those at high risk are more precisely targeted. Although dental (especially periodontal) infections have been recently identified as independent risk factors for CAD, current evidence is insufficient to justify treatment of such infections to arrest or reverse CAD or other systemic conditions (e.g., diabetes mellitus, stroke or adverse outcomes of pregnancies). PMID- 11911815 TI - Is there an association between edentulism and nutritional state? AB - Edentulous people have difficulty chewing foods that are hard or tough in texture, even when wearing well-made dentures. These individuals typically modify their diets to compensate for loss of oral function. This practice leads to the question of whether the diet of edentulous individuals is adequate to maintain good general health. This overview summarizes articles that describe the changes in diet associated with edentulism. Such changes include reductions in fruits, vegetables, meats and other hard-to-chew foods and are associated with compromised nutrition. The evidence suggests that edentulous individuals lack specific nutrients and, as a result, may be at risk for various health disorders. The authors have recently shown that mandibular prostheses supported by only 2 implants may significantly improve nutritional status in edentulous patients. PMID- 11911816 TI - Periodontal health and systemic disorders. AB - Recent studies in periodontal medicine suggest a mild to moderate association between human periodontal disease and certain systemic disorders such as diabetes mellitus, pneumonia, heart disease and preterm birth. The latest evidence, presented at a symposium entitled Periodontal Health and Systemic Disorders, sponsored by the University of Western Ontario School of Dentistry, showed that indeed such an association is likely. New data suggest that this association is not indicated by traditional clinical signs of periodontal disease but rather by a cluster of host immune and inflammatory mediators. The coming era of periodontal medicine based upon molecular criteria will affect the future of periodontal diagnosis, treatment and professional practice. PMID- 11911817 TI - Immune response to Salmonella: location, location, location? AB - Successful immunity against Salmonella infections is dependent on the generation of CD4(+) T helper cells and to a lesser extent on antibody production and CD8(+) T cells. The cells within the lymphatic tissue of the gut are likely to be central for the orchestration of a proper and rapid response. The anatomical restriction of the pathogen may also determine the distribution of effector cells. In this issue of Immunity, McSorley et al. address both of these processes using identifiable CD4 T cells that are specific for Salmonella typhimurium. Such cells localize to the Peyer's patches of the small intestine when the bacteria are delivered orally. PMID- 11911819 TI - Vav1 controls integrin clustering and MHC/peptide-specific cell adhesion to antigen-presenting cells. AB - Integrin-mediated adhesion is essential for the formation of stable contacts between T cells and antigen-presenting cells (APCs). We show that Vav1 controls integrin-mediated adhesion of thymocytes and T cells to ECM proteins and ICAM1 following TCR stimulation. In a peptide-specific system, Vav1 is required for T cell adhesion to peptide-loaded APCs. Intriguingly, TCR-induced cell adhesion and aggregation of integrins occurs independent of WASP. Whereas LFA-1 and actin caps colocalize in wasp(-/-) T cells in response to TCR stimulation, loss of WASP uncouples TCR caps from actin patches. Our data reveal a novel role for Vav1 and WASP in the regulation of TCR-induced integrin clustering and cell adhesion and show that integrin and TCR clustering are controlled by distinct pathways. PMID- 11911820 TI - A T cell receptor CDR3beta loop undergoes conformational changes of unprecedented magnitude upon binding to a peptide/MHC class I complex. AB - The elongated complementary-determining region (CDR) 3beta found in the unliganded KB5-C20 TCR protrudes from the antigen binding site and prevents its docking onto the peptide/MHC (pMHC) surface according to a canonical diagonal orientation. We now present the crystal structure of a complex involving the KB5 C20 TCR and an octapeptide bound to the allogeneic H-2K(b) MHC class I molecule. This structure reveals how a tremendously large CDR3beta conformational change allows the KB5-C20 TCR to adapt to the rather constrained pMHC surface and achieve a diagonal docking mode. This extreme case of induced fit also shows that TCR plasticity is primarily restricted to CDR3 loops and does not propagate away from the antigen binding site. PMID- 11911821 TI - Precursors of functional MHC class I- or class II-restricted CD8alphaalpha(+) T cells are positively selected in the thymus by agonist self-peptides. AB - The origin and specificity of alphabeta TCR(+) T cells that express CD8alphaalpha have been controversial issues. Here we provide direct evidence that precursors of functional CD8alphaalpha T cells are positively selected in the thymus in the presence of agonist self-peptides. Like conventional positive selection, this agonist selection process requires functional TCR alpha-CPM, whereas it is independent of CD8beta expression. Furthermore, CD8alphaalpha expression on mature, agonist-selected T cells does not imply selection by MHC class I, and CD8alphaalpha(+) T cells can be either class I or class II restricted. Our data define a distinct agonist-dependent, positive selection process in the thymus, and they suggest a function for CD8alphaalpha distinct from the conventional TCR coreceptor function of CD8alphabeta or CD4. PMID- 11911822 TI - Tracking salmonella-specific CD4 T cells in vivo reveals a local mucosal response to a disseminated infection. AB - A novel adoptive transfer system was used to track the fate of naive Salmonella specific CD4 T cells in vivo. These cells showed signs of activation in the Peyer's patches as early as 3 hr after oral infection. The activated CD4 T cells then produced IL-2 and proliferated in the T cell areas of these tissues before migrating into the B cell-rich follicles. In contrast, Salmonella-specific CD4 T cells were not activated in the spleen and very few of these cells migrated to the liver, despite the presence of bacteria in both organs. These results show that the T cell response to pathogenic Salmonella infection is localized to the gut-associated lymphoid tissue and does not extend efficiently to the major sites of late infection. PMID- 11911823 TI - FcgammaRI-deficient mice show multiple alterations to inflammatory and immune responses. AB - The inactivation of the mouse high-affinity IgG Fc receptor FcgammaRI resulted in a wide range of defects in antibody Fc-dependent functions. These studies showed the primary importance of FcgammaRI in endocytosis of monomeric IgG, kinetics, and extent of phagocytosis of immune complexes, in macrophage-based ADCC, and in immune complex-dependent antigen presentation to primed T cells. In the absence of FcgammaRI, antibody responses were elevated, implying the removal of a control point by the deletion of FcgammaRI. In addition, FcR-gamma chain-deficient mice were found to express partially functional FcgammaRI. Thus, FcgammaRI is an early participant in Fc-dependent cell activation and in the development of immune responses. PMID- 11911824 TI - FcgammaRI (CD64) contributes substantially to severity of arthritis, hypersensitivity responses, and protection from bacterial infection. AB - The high-affinity receptor for IgG, FcgammaRI, shares its capacity to bind IgG2a immune complexes (IgG2a-IC) with the low-affinity receptor FcgammaRIII and complement factors, hampering the definition of its biological role. Moreover, in vivo, FcgammaRI is occupied by monomeric IgG2a, reducing its accessibility to newly formed IgG2a-IC. By using a variety of FcgammaR(-/-) mice, we demonstrate that in the absence of FcgammaRI, the IgG2a-IC-induced cellular processes of phagocytosis, cytokine release, cellular cytotoxicity, and antigen presentation are impaired. FcgammaRI(-/-) mice showed impaired hypersensitivity responses, strongly reduced cartilage destruction in an arthritis model, and impaired protection from a bacterial infection. We conclude that FcgammaRI contributes substantially to a variety of IgG2a-IC-dependent immune functions and immunopathological responses. PMID- 11911825 TI - CD40L blockade prevents autoimmune diabetes by induction of bitypic NK/DC regulatory cells. AB - Systemic treatment with antibody to CD40 ligand (aCD40L) can prevent autoimmunity and transplant rejection in several animal models and is currently under evaluation in clinical trials. While it is known that aCD40L administration inhibits expansion and effector functions of aggressive T cells, it is still unclear whether additional regulatory mechanisms are operative. Here we demonstrate that a single episode of CD40L blockade during development of the autoaggressive immune response completely prevented autoimmune disease in the RIP LCMV mouse model for virally induced type 1 diabetes. Interestingly, protection could be transferred by a highly potent, bitypic cell population sharing phenotypic and functional properties of both natural killer (NK) and dendritic cells (DC). Furthermore, protection of prediabetic recipients was autoantigen specific and did not result in generalized immunosuppression. The origin, function, and therapeutic potential of these bitypic NK/DC regulatory cells is discussed. PMID- 11911826 TI - Cytotoxic cell granule-mediated apoptosis: perforin delivers granzyme B-serglycin complexes into target cells without plasma membrane pore formation. AB - The mechanism underlying perforin (PFN)-dependent delivery of apoptotic granzymes during cytotoxic cell granule-mediated death remains speculative. Granzyme B (GrB) and perforin were found to coexist as multimeric complexes with the proteoglycan serglycin (SG) in cytotoxic granules, and cytotoxic cells were observed to secrete exclusively macromolecular GrB-SG. Contrary to the view that PFN acts as a gateway for granzymes through the plasma membrane, monomeric PFN and, strikingly, PFN-SG complexes were shown to mediate cytosolic delivery of macromolecular GrB-SG without producing detectable plasma membrane pores. These results indicate that granule-mediated apoptosis represents a phenomenon whereby the target cell perceives granule contents as a multimeric complex consisting of SG, PFN, and granzymes, which are, respectively, the scaffold, translocator, and targeting/informational components of this modular delivery system. PMID- 11911827 TI - In the absence of IL-12, CD4(+) T cell responses to intracellular pathogens fail to default to a Th2 pattern and are host protective in an IL-10(-/-) setting. AB - IL-12-deficient mice exposed to nonlethal infections with intracellular pathogens or repeatedly immunized with a pathogen extract developed lowered but nevertheless substantial numbers of IFN-gamma(+) CD4(+) T cells compared to those observed in wild-type animals. Moreover, the CD4(+) responses in these knockout animals failed to default to a Th2 pattern. The protective efficacy of the Th1 cells developing in an IL-12-deficient setting was found to be limited by IL-10 since mice doubly deficient in IL-10 and IL-12 survived, while animals deficient in IL-12 alone succumbed to pathogen challenge. In contrast to IL-12 knockout mice, MyD88-deficient animals exposed to a Th1 microbial stimulus developed a pure Th2 response, arguing that this signaling element plays a more critical function than IL-12 in determining pathogen-induced CD4 polarization. PMID- 11911828 TI - Phosphoinositide 3-kinase gamma is an essential amplifier of mast cell function. AB - Mast cells are key regulators in allergy and inflammation, and release histamine upon clustering of their IgE receptors. Here we demonstrate that murine mast cell responses are exacerbated in vitro and in vivo by autocrine signals through G protein-coupled receptors (GPCRs) and require functional phosphoinositide 3 kinase gamma (PI3Kgamma). Adenosine, acting through the A(3) adenosine receptor (A(3)AR) as well as other agonists of G(alphai)-coupled GPCRs, transiently increased PtdIns(3,4,5)P(3) exclusively via PI3Kgamma. PI3Kgamma-derived PtdIns(3,4,5)P(3) was instrumental for initiating a sustained influx of external Ca(2+) and degranulation. Mice lacking PI3Kgamma did not form edema after intradermal injection of adenosine and when challenged by passive systemic anaphylaxis. PI3Kgamma thus relays inflammatory signals through various G(i) coupled receptors and is central to mast cell function. PMID- 11911829 TI - Redundant and unique roles of two enhancer elements in the TCRgamma locus in gene regulation and gammadelta T cell development. AB - Many mammalian genes, including those encoding antigen receptors, contain more than one enhancer element. Deleting one element often does not prevent expression, but functional redundancy has never been directly demonstrated by gene targeting of multiple elements. We demonstrate that simultaneous deletion of two enhancer/LCR-like elements in the TCR Cgamma1 cluster, HsA and 3'E(Cgamma1), severely diminishes TCRgamma transcription, selectively impairs development of gammadelta thymocyte subsets, but only modestly reduces TCRgamma gene rearrangement, while deletion of each element separately has little effect. In contrast to these results in thymocytes, deletion of HsA alone reduces transcription of one Vgamma gene specifically in peripheral gammadelta T cells. Thus, the two elements exhibit functional redundancy in thymocytes but also have unique functions in other settings. PMID- 11911830 TI - Beta1 integrin is not essential for hematopoiesis but is necessary for the T cell dependent IgM antibody response. AB - Several experimental evidences suggested that beta1 integrin-mediated adhesion of hematopoietic stem cells (HSC) is important for their function in the bone marrow (BM). Using induced deletion of the beta1 integrin gene restricted to the hematopoietic system, we show that beta1 integrin is not essential for HSC retention in the BM, hematopoiesis, and trafficking of lymphocytes. However, immunization with a T cell-dependent antigen resulted in virtually no IgM production and an increased secretion of IgG in mutant mice, while the response to a T cell-independent type 2 antigen showed decreases in both IgM and IgG. These data suggest that beta1 integrins are necessary for the primary IgM antibody response. PMID- 11911831 TI - TL1A is a TNF-like ligand for DR3 and TR6/DcR3 and functions as a T cell costimulator. AB - DR3 is a death domain-containing receptor that is upregulated during T cell activation and whose overexpression induces apoptosis and NF-kappaB activation in cell lines. Here we show that an endothelial cell-derived TNF-like factor, TL1A, is a ligand for DR3 and decoy receptor TR6/DcR3 and that its expression is inducible by TNF and IL-1alpha. TL1A induces NF-kappaB activation and apoptosis in DR3-expressing cell lines, while TR6-Fc protein antagonizes these signaling events. Interestingly, in T cells, TL1A acts as a costimulator that increases IL 2 responsiveness and secretion of proinflammatory cytokines both in vitro and in vivo. Our data suggest that interaction of TL1A with DR3 promotes T cell expansion during an immune response, whereas TR6 has an opposing effect. PMID- 11911833 TI - Alterations in the expression of cytochrome c oxidase subunits in doxorubicin resistant leukemia K562 cells. AB - Doxorubicin (DOX), a widely used antitumoral drug, induces numerous modifications in sensitive cells, interacting with nuclear and mitochondrial DNA. In previous studies achieved in two K562 DOX-resistant sublines (K562/0.2R and K562/0.5R), we have shown stable mitochondrial damage comparatively with sensitive parental cells, such as decrease of cytochrome c oxidase activity (COX; EC 1.9.3.1) and cytochrome aa3 content. In order to explain these data, we have studied several COX genes and their expression, in relationship with altered COX activity and multidrug resistance (MDR) phenotype. We have observed a lower expression of the catalytic subunits COX I and II in MDR sublines, which was neither related to mutations in the corresponding mitochondrial genes, nor to a reduced transcription rate. In contrast, we have noticed an increase in both MDR K562 variants, in the mRNA expression of the catalytic subunit COX III, related to an increase in the half-life of these transcripts. Moreover, the doxorubicin resistance phenotype in K562 cells was accompanied by modifications of the expression and steady-state mRNA levels of several nuclear-encoded regulatory COX subunits. Thus, doxorubicin-resistant K562 cells represent an interesting model to study stable modifications concomitant to MDR phenotype. Our results seem to indicate compensatory mechanisms which highlight the complexity of regulatory systems of COX enzyme, involving coordinate regulation of both nuclear and mitochondrial subunit expression. PMID- 11911832 TI - Selective cyclooxygenase-2 (COX-2) inhibitors and potential risk of cardiovascular events. AB - Selective cyclooxygenase-2 (COX-2) inhibitors were developed as a response to the gastrointestinal toxicity of conventional nonsteroidal anti-inflammatory agents (NSAIDs). However, COX-2 inhibitors decrease vascular prostacyclin (PGI(2)) production and may disrupt the homeostatic mechanisms that limit the effects of platelet activation. Basic and clinical data raise concerns about a potential prothrombotic effect of this class of drugs. The widespread popularity of these agents mandates their prospective evaluation in patients with cardiovascular diseases or who are at risk for cardiovascular events. PMID- 11911834 TI - Novel phenothiazine antimalarials: synthesis, antimalarial activity, and inhibition of the formation of beta-haematin. AB - We report the synthesis of a series of novel phenothiazine compounds that inhibit the growth of both chloroquine-sensitive and chloroquine-resistant strains of Plasmodium falciparum. We found that the antimalarial activity of these phenothiazines increased with an increase in the number of basic groups in the alkylamino side chain, which may reflect increased uptake into the parasite food vacuole or differences in the toxicities of individual FP-drug complexes. We have examined the ability of the parent phenothiazine, chlorpromazine, and some novel phenothiazines to inhibit the formation of beta-haematin. The degree of antimalarial potency was loosely correlated with the efficacy of inhibition of beta-haematin formation, suggesting that these phenothiazines exert their antimalarial activities in a manner similar to that of chloroquine, i.e. by antagonizing the sequestration of toxic haem (ferriprotoporphyrin IX) moieties within the malaria parasite. Chlorpromazine is an effective modulator of chloroquine resistance; however, the more potent phenothiazine derivatives were more active against chloroquine-sensitive parasites than against chloroquine resistant parasites and showed little synergy of action when used in combination with chloroquine. These studies point to structural features that may determine the antimalarial activity and resistance modulating potential of weakly basic amphipaths. PMID- 11911835 TI - Increased rate of glutathione synthesis from cystine in drug-resistant MCF-7 cells. AB - The rate of glutathione synthesis was determined in drug-sensitive and -resistant MCF-7 cells by monitoring the rate of label uptake from [3,3'-13C(2)]-cystine using NMR spectroscopy and mass spectrometry. Compared with the wild-type human mammary adenocarcinoma cell line (MCF-7wt), the isotope incorporation rate was increased 1.6-, 2.4-, and 5.3-fold in the etoposide-resistant MCF-7 cell line (MCF-7vp), doxorubicin-resistant MCF-7 cell line (MCF-7adr), and 4 hydroperoxycyclophosphamide-resistant MCF-7 cell line (MCF-7hc), respectively. The increase in glutathione metabolism in the MCF-7hc line correlated with steady state levels as determined by biochemical assay. In contrast, both the MCF-7vp and MCF-7adr lines showed increased metabolic synthesis of glutathione but displayed lower steady-state levels compared with the MCF-7wt line. The increased synthetic rates of all resistant lines reflected, in part, contributions from the increased activities of both gamma-glutamyltranspeptidase and gamma glutamylcysteine synthetase. These results emphasize the importance of monitoring glutathione metabolic rates, rather than steady-state levels of enzymes or substrates, for assessing drug resistance. PMID- 11911836 TI - Effects of sanguiin H-6, a component of Sanguisorbae Radix, on lipopolysaccharide stimulated nitric oxide production. AB - The present study was conducted to evaluate the effect of sanguiin H-6, a component of Sanguisorbae Radix, on the production of nitric oxide (NO), using macrophages activated by lipopolysaccharide (LPS). Sanguiin H-6 inhibited nitrite production, taken as an index for NO, in a concentration-dependent fashion. This compound decreased inducible NO synthase (iNOS) activity, with the inhibitory effect at a concentration of 25 microM being equal to that of the known iNOS inhibitor aminoguanidine at 50 microM. However, unlike aminoguanidine, sanguiin H 6 was associated with improved cell viability. Reverse transcription-polymerase chain reaction analysis revealed that the expression of iNOS mRNA in activated macrophages was suppressed by sanguiin H-6 in a concentration-dependent manner. In addition, sanguiin H-6 even at a low concentration showed a clear scavenging effect on the NO generated from sodium nitroprusside (an NO donor). These findings indicate that not only does sanguiin H-6 act directly as an NO scavenger, but it also inhibits NO production in LPS-activated macrophages by the concomitant inhibition of iNOS mRNA induction and enzyme activity. PMID- 11911837 TI - Association of dopamine D(3) receptors with actin-binding protein 280 (ABP-280). AB - Proteins that bind to G protein-coupled receptors have been identified as regulators of receptor localization and signaling. In our previous studies, a cytoskeletal protein, actin-binding protein 280 (ABP-280), was found to associate with the third cytoplasmic loop of dopamine D(2) receptors. In this study, we demonstrate that ABP-280 also interacts with dopamine D(3) receptors, but not with D(4) receptors. Similar to the dopamine D(2) receptor, the D(3)/ABP-280 association is of signaling importance. In human melanoma M2 cells lacking ABP 280, D(3) receptors were unable to inhibit forskolin-stimulated cyclic AMP (cAMP) production significantly. D(4) receptors, however, exhibited a similar degree of inhibition of forskolin-stimulated cAMP production in ABP-280-deficient M2 cells and ABP-280-replent M2 subclones (A7 cells). Further experiments revealed that the D(3)/ABP-280 interaction was critically dependent upon a 36 amino acid carboxyl domain of the D(3) receptor third loop, which is conserved in the D(2) receptor but not in the D(4) receptor. Our results demonstrate a subtype-specific regulation of dopamine D(2)-family receptor signaling by the cytoskeletal protein ABP-280. PMID- 11911838 TI - Phentermine inhibition of recombinant human liver monoamine oxidases A and B. AB - Recent studies with rat tissue preparations have suggested that the anorectic drug phentermine inhibits serotonin degradation by inhibition of monoamine oxidase (MAO) A with a K(I) value of 85-88 microM, a potency suggested to be similar to that of other reversible MAO inhibitors (Ulus et al., Biochem Pharmacol 2000;59:1611-21). Since there are known differences between rats and humans in substrate and inhibitor specificities of MAOs, the interactions of phentermine with recombinant human purified preparations of MAO A and MAO B were determined. Human MAO A was competitively inhibited by phentermine with a K(I) value of 498+/-60 microM, a value approximately 6-fold weaker than that observed for the rat enzyme. Phentermine was also observed to be a competitive inhibitor of recombinant human liver MAO B with a K(I) value of 375+/-42 microM, a value similar to that observed with the rat enzyme (310-416 microM). In contrast to the behavior with rat tissue preparations, no slow time-dependent behavior was observed for phentermine inhibition of purified soluble human MAO preparations. Difference absorption spectral studies showed similar perturbations of the covalent FAD moieties of both human MAO A and MAO B, which suggests a similar mode of binding in both enzymes. These data suggest that phentermine inhibition of human MAO A (or of MAO B) is too weak to be of pharmacological relevance. PMID- 11911839 TI - p53-Independent induction of Fas and apoptosis in leukemic cells by an adenosine derivative, Cl-IB-MECA. AB - A(3) adenosine receptor (A(3)AR) agonists have been reported to influence cell death and survival. The effects of an A(3)AR agonist, 1-[2-chloro-6-[[(3 iodophenyl)methyl]amino]-9H-purin-9-yl]-1-deoxy-N-methyl-beta-D-ribofuranonamide (Cl-IB-MECA), on apoptosis in two human leukemia cell lines, HL-60 and MOLT-4, were investigated. Cl-IB-MECA (> or =30 microM) increased the apoptotic fractions, as determined using fluorescence-activated cell sorting (FACS) analysis, and activated caspase 3 and poly-ADP-ribose-polymerase. Known messengers coupled to A(3)AR (phospholipase C and intracellular calcium) did not seem to play a role in the induction of apoptosis. Neither dantrolene nor BAPTA AM affected the Cl-IB-MECA-induced apoptosis. Cl-IB-MECA failed to activate phospholipase C in HL-60 cells, while UTP activated it through endogenous P2Y(2) receptors. Induction of apoptosis during a 48hr exposure to Cl-IB-MECA was not prevented by the A(3)AR antagonists [5-propyl-2-ethyl-4-propyl-3 (ethylsulfanylcarbonyl)-6-phenylpyridine-5-carboxylate] (MRS 1220) or N-[9-chloro 2-(2-furanyl)[1,2,4]triazolo[1,5-c]quinazolin-5-yl]benzeneacetamide (MRS 1523). Furthermore, higher concentrations of MRS 1220, which would also antagonize A(1) and A(2A) receptors, were ineffective in preventing the apoptosis. Although Cl-IB MECA has been shown in other systems to cause apoptosis through an A(3)AR mediated mechanism, in these cells it appeared to be an adenosine receptor independent effect, which required prolonged incubation. In both HL-60 and MOLT-4 cells, Cl-IB-MECA induced the expression of Fas, a death receptor. This induction of Fas was not dependent upon p53, because p53 is not expressed in an active form in either HL-60 or MOLT-4 cells. Cl-IB-MECA-induced apoptosis in HL-60 cells was augmented by an agonistic Fas antibody, CH-11, and this increase was suppressed by the antagonistic anti-Fas antibody ZB-4. Therefore, Cl-IB-MECA induced apoptosis via a novel, p53-independent up-regulation of Fas. PMID- 11911840 TI - Fas/CD95-mediated apoptosis in human glioblastoma cells: a target for sensitisation to topoisomerase I inhibitors. AB - The expression of the death receptor Fas/CD95 is cell type-specific and can be modulated by different cytotoxic treatments. In spite of a frequent expression of Fas/CD95 in high-grade gliomas, these tumours are typically refractory to conventional therapy. Using a human glioblastoma cell line (GBM), we explored the possibility of modulating susceptibility to Fas/CD95-mediated apoptosis following cytotoxic treatment. GBM cells were sensitive to the antiproliferative effects of topoisomerase I inhibitors (topotecan and a novel lipophilic analog CPT83) and taxol, less sensitive to cisplatin and, in any case, rather resistant to drug induced apoptosis. This pattern of cellular response was consistent with p53 mutation. GBM cells expressed low levels of Fas/CD95, which was associated with low susceptibility to antibody-stimulated Fas/CD95-mediated apoptosis. A significant up-regulation of Fas/CD95 expression was detected after exposure to topotecan and CPT83, whereas cisplatin induced a low increase and taxol did not modify Fas/CD95 expression. In addition, after treatment with topoisomerase I inhibitors, the up-regulation of Fas/CD95 resulted in an increased susceptibility of GBM cells to antibody-stimulated Fas/CD95-mediated apoptosis, while no synergistic effects were detected after treatment with cisplatin or taxol. Our data suggest that Fas/CD95 up-regulation can be a common response to DNA damage, whereas sensitisation to Fas/CD95-mediated apoptosis appears to be dependent on the type of DNA damage and on the pathway of cellular response. The observed effects might have important therapeutic implications for the design of novel therapeutic strategies in the treatment of malignant gliomas. PMID- 11911841 TI - Substrate specificity for rat cytochrome P450 (CYP) isoforms: screening with cDNA expressed systems of the rat. AB - In this study, we performed a screening of the specificities of rat cytochrome P450 (CYP) isoforms for metabolic reactions known as the specific probes of human CYP isoforms, using 13 rat CYP isoforms expressed in baculovirus-infected insect cells or B-lymphoblastoid cells. Among the metabolic reactions studied, diclofenac 4-hydroxylation (DFH), dextromethorphan O-demethylation (DMOD) and midazolam 4-hydroxylation were specifically catalyzed by CYP2C6, CYP2D2 and CYP3A1/3A2, respectively. These results suggest that diclofenac 4-hydroxylation, dextromethorphan O-demethylation and midazolam 4-hydroxylation are useful as catalytic markers of CYP2C6, CYP2D2 and CYP3A1/3A2, respectively. On the other hand, phenacetin O-deethylation and 7-ethoxyresorufin O-deethylation were catalyzed both by CYP1A2 and by CYP2C6. Benzyloxyresorufin O-dealkylation and pentoxyresorufin O-dealkylation were also catalyzed by CYP1A2 in addition to CYP2B1. Bufuralol 1'-hydroxylation was extensively catalyzed by CYP2D2 but also by CYP2C6 and CYP2C11. p-Nitrophenol 2-hydroxylation and chlorzoxazone 6 hydroxylation were extensively catalyzed by CYP2E1 but also by CYP1A2 and CYP3A1. Therefore, it is necessary to conduct further study to clarify whether these activities in rat liver microsomes are useful as probes of rat CYP isoforms. In contrast, coumarin 7-hydroxylation and S- and R-mephenytoin 4'-hydroxylation did not show selectivity toward any isoforms of rat CYP studied. Therefore, activities of coumarin 7-hydroxylation and S- and R-mephenytoin 4'-hydroxylation are not able to be used as catalytic probes of CYP isoforms in rat liver microsomes. These results may provide useful information regarding catalytic probes of rat CYPs for studies using rat liver microsomal samples. PMID- 11911842 TI - A doxorubicin-CNGRC-peptide conjugate with prodrug properties. AB - There is increasing interest in the exploitation of molecular addresses for the targeting of tumor imaging or therapeutic agents. A recent study demonstrated anticancer activity in human xenografts of doxorubicin (DOX)-peptide conjugates targeted to the tumor vascular endothelium, among them DOX coupled to the cyclic pentapeptide CNGRC [Science 279 (1998) 377]. In order to learn more about the mechanism of action of this type of DOX-peptide conjugates, we have studied the interaction of DOX-CNGRC with primary human umbilical cord vein endothelial cells (HUVEC) and tumor cells under defined in vitro conditions. We used a DOX conjugate, in which the cyclic CNGRC peptide, for which an in vivo endothelial address has recently been identified as aminopeptidase N (APN)/CD13, has been coupled via a hydrolysable spacer to the C-14 anthracycline-side chain. First we determined that the t(1/2) of DOX-CNGRC conjugate in human blood was 442 min (at 37 degrees ) allowing sufficient time for endothelial targeting when administered i.v. When cultured cells were exposed for 30 min to DOX-CNGRC a more cytoplasmic localization of fluorescent drug was seen when compared to DOX exposure and intracellular DOX-CNGRC was identified after extraction from the cells. This revealed differences in the cellular uptake process of the conjugate compared to DOX. The antiproliferative effect of DOX-CNGRC was determined by 30 min exposure in medium with a high protein content in order to mimick the in vivo targeting situation. In this medium, the IC(50) was 1.1 microM for highly CD13 expressing HT-1080, 1.45 microM for CD13 negative SK-UT-1 sarcoma cells and 6.5 microM for CD13 positive HUVEC. The IC(50) of DOX for these cells were 1.0, 2.0 and 7.3 microM, respectively. Although DOX-CNGRC inhibited the peptidase activity of CD13 up to 50%, our data do not favor an important role for the enzyme inhibition in the cytotoxic effect of the conjugate. The antitumor activity was tested in nude mice bearing human ovarian cancer xenografts (OVCAR-3). A weekly i.v. administration (3mg/kg DOX-equivalent, 3x) showed a minor (40%) growth delay, which does not indicate efficacy better than that expected for free DOX. In conclusion, this study indicates that the antiproliferative and anti-angiogenic effects of DOX-CNGRC as reported before, are likely caused by the cytostatic effects of intracellularly released parent drug DOX, independent of CD13 expression/activity. More research is needed to identify the optimal specific chemical configuration of DOX-peptide conjugates for in vivo targeting and receptor-mediated cellular uptake. PMID- 11911844 TI - Reduction of acute photodamage in skin by topical application of a novel PARP inhibitor. AB - The ultraviolet (UV) components of sunlight induce damage to the DNA in skin cells, which is considered to be the initiating step in the harmful biological effects of UV radiation. Repair of DNA damage results in the formation of single strand DNA breaks, which activate the nuclear poly(ADP-ribose) polymerase (PARP). Overactivation of PARP worsens the oxidative cell damage and impairs the energy metabolism, raising the possibility that moderation of PARP activation following DNA damage may protect skin cells from UV radiation. The topical effects of the novel PARP inhibitor O-(3-pyperidino-2-hydroxy-1-propyl) pyridine-3-carboxylic acid amidoxime monohydrochloride (BGP-15M) were investigated on UV-induced skin damage in a hairless mouse model. For evaluation of the UV-induced acute photodamage to the skin and the potential protective effect of BGP-15M, DNA injury was detected by measuring the formation of single-strand DNA breaks and counting the resulting sunburn (apoptotic) cells. The ADP-ribosylation of PARP was assessed by Western blot analysis and then quantified. In addition, the UV induced immunosuppression was investigated by the immunostaining of tumor necrosis factor alpha and interleukin-10 expressions in epidermal cells. The signs of inflammation were examined clinically and histochemically. Besides its primary effect in decreasing the activity of nuclear PARP, topically applied BGP 15M proved to be protective against solar and artificial UV radiation-induced acute skin damage. The DNA injury was decreased (P<0.01). An inhibition of immunosuppression was observed by down-regulation of the epidermal production of cytokines IL-10 and TNFalpha. In the mouse skin, clinical or histological signs of UV-induced inflammation could not be observed. These data suggest that BGP-15M directly interferes with UV-induced cellular processes and modifies the activity of PARP. The effects provided by topical application of the new PARP-regulator BGP-15M indicate that it may be a novel type of agent in photoprotection of the skin. PMID- 11911843 TI - Selective dopaminergic neurotoxicity of isoquinoline derivatives related to Parkinson's disease: studies using heterologous expression systems of the dopamine transporter. AB - Endogenous isoquinoline (IQ) derivatives structurally related to the selective dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its active metabolite 1-methyl-4-phenylpyridine (MPP(+)) may contribute to dopaminergic neurodegeneration in Parkinson's disease. We addressed the importance of the DAT molecule for selective dopaminergic toxicity by testing the differential cytotoxicity of 22 neutral and quaternary compounds from three classes of isoquinoline derivatives (3, IQs; 4,3,4-dihydroisoquinolines and 15, 1,2,3,4-tetrahydroisoquinolines) as well as MPP(+) in non-neuronal and neuronal heterologous expression systems of the DAT gene (human embryonic kidney HEK-293 and mouse neuroblastoma Neuro-2A cells, respectively). Cell death was estimated using the MTT assay and the Trypan blue exclusion method. Nine isoquinolines and MPP(+) showed general cytotoxicity in both parental cell lines after 72hr with half-maximal toxic concentrations (TC(50) values) in the micromolar range. The rank order of toxic potency was: papaverine>salsolinol=tetrahydropapaveroline=1 benzyl-TIQ=norsalsolinol>tetrahydropapaverine>2[N]-methyl-salsolinol>2[N]-methyl norsalsolinol>2[N]-Me-IQ(+)=MPP(+). Besides MPP(+), only the 2[N]-methylated compounds 2[N]-methyl-IQ(+), 2[N]-methyl-norsalsolinol and 2[N]-methyl-salsolinol showed enhanced cytotoxicity in both DAT expressing cell lines with 2- to 14-fold reduction of TC(50) values compared to parental cell lines. The rank order of selectivity in both cell systems was: MPP(+)>>2[N]-Me-IQ(+)>2[N]-methyl norsalsolinol=2[N]-methyl-salsolinol. Our results suggest that 2[N]-methylated isoquinoline derivatives structurally related to MPTP/MPP(+) are selectively toxic to dopaminergic cells via uptake by the DAT, and therefore may play a role in the pathogenesis of Parkinson's disease. PMID- 11911845 TI - Human cytochrome P450s involved in the metabolism of 9-cis- and 13-cis-retinoic acids. AB - The purpose of this work was to identify the principal human cytochrome P450s (CYPs) involved in the metabolism of the retinoic acid (RA) isomers, 9-cis- and 13-cis-RA, by using a combination of techniques including human liver microsomes (correlation of activity and inhibition), and lymphoblast microsomes expressing a single CYP. Concerning the 9-cis-RA, 4-OH- and 4-oxo-9-cis-RA were formed with human liver microsomes, and their formation correlated with activities linked to CYPs 3A4/5, 2B6, 2C8, 2A6, and 2C9. The use of lymphoblast microsomes expressing a single human CYP identified CYPs 2C9>2C8>3A7 as the most active in the formation of 4-OH-9-cis-RA. With regard to 13-cis-RA, specific P450 activities linked to CYPs 2B6, 2C8, 3A4/5, and 2A6 were correlated with the formation of 4 OH- and 4-oxo-13-cis-RA. Microsomes expressing a single CYP identified CYPs 3A7, 2C8, 4A11, 1B1, 2B6, 2C9, 2C19, 3A4 (decreasing activity) in the formation of 4 OH-13-cis-RA. The use of CYP-specific inhibitors in human liver microsomes disclosed that the formation of the 4-OH-9-cis-RA was best inhibited by sulfaphenazole (72%) and quercetin (66%), whereas ketoconazole and troleandomycin inhibited its formation by 55 and 38%, respectively; the formation of 4-OH-13-cis RA was best inhibited by troleandomycin (54%) and ketoconazole (46%), whereas quercetin was a weak inhibitor (14%). In conclusion, adult human CYPs 2C9, 2C8, 3A4 have been identified as active in the 9-cis-RA metabolism, whereas CYPs 3A4 and 2C8 were active in 13-cis-RA metabolism. The fetal form CYP3A7 was also identified as very active in either 9-cis- or 13-cis-RA metabolism. The role of these human CYPs in the biological response or resistance to RA isomers remains to be determined. PMID- 11911846 TI - Pharmacological characterization of the ATP-dependent low K(m) guanosine 3',5' cyclic monophosphate (cGMP) transporter in human erythrocytes. AB - The efflux pump for cGMP has been shown to be an ATP-energized multiorganic anion transporter. The present study was performed to extend the knowledge of the pharmacological characteristics of this efflux pump. Inside-out vesicles prepared from fresh blood were incubated with [3H]-cGMP (1 microM) with or without various concentrations of competitors for 120min at 37 degrees. The tested compounds could be divided in four groups: one with high affinity (K(i) < 5 microM), a second with moderate affinity (K(i): 5-50 microM), a third with low affinity (K(i): 0.1-5mM) and the fourth with extremely low or no affinity at all. With the mean K(i)-values given in parenthesis, the high affinity group consisted of mifepristone (0.2 microM), zaprinast (0.35 microM), dipyridamole (0.35 microM), estradiol 3-beta-glucuronide (0.42 microM), genistein (0.43 microM), estradiol 17 beta-glucuronide (0.47 microM), onapristone (1.3 microM), progesterone (1.7 microM) and sildenafil (3.6 microM). The inhibitors with medium affinity were estradiol (8 microM), sulfinpyrazone (13 microM), daunorubicin (23 microM), megestrol acetate (26 microM), doxorubicin (28 microM), 6-thioguanine (28 microM) and 6-thioguanosine-5'-monophosphate (32 microM). The low affinity group comprised 6-TIMP (220 microM), 6-methylmercaptopurine (MMP) (220 microM), vincristine (270 microM), medroxyprogesterone (680 microM), para-aminohippurate (PAH) (1.9mM) and taurocholate (2.2mM). No or minimal effect was seen in the presence of 6-mercaptopurine (6-MP), methotrexate, 9-(2 phosphonylmethoxyethyl)adenine and mitoxantrone. The cGMP transporter had a unique pharmacological profile, different from that of MRP1, but with some characteristics in common with MRP4 and MRP5. PMID- 11911847 TI - Jolkinolide B induces neuroendocrine differentiation of human prostate LNCaP cancer cell line. AB - Euphorbia fischeriana is a Chinese herbal medicine which has been reported to possess chemotherapeutic effects, yet the underlying mechanism is unclear. In order to understand its possible anti-tumor property, we have isolated a number of chemical compounds from the roots of this plant [Phytochemistry 52 (1999) 117] and studied their in vitro effects by using human prostate LNCaP cancer cell line. Among the six compounds tested, jolkinolide B exhibited the most potent anti-proliferative activity (IC(50)=12.5 microg/mL=40 microM) and it inhibited DNA synthesis by down-regulating bromodeoxyuridine (BrdU) incorporation in LNCaP cells in a dose-dependent manner. Jolkinolide B, at concentrations up to 25 microg/mL, induced G1 arrest and neuroendocrine differentiation of LNCaP cells. Immunoblotting analysis confirmed the increased expression of neuroendocrine markers, keratin 8/18 (K8/18) and neuron specific enolase (NSE), in these cells. Apoptotic bodies and DNA fragmentation were observed by fluorescence microscopy and flow cytometry when the cells were exposed to a concentration higher than 25 microg/mL jolkinolide B. Taken all data together, jolkinolide B seems to play a role in the regulation of proliferation, differentiation, and apoptosis of LNCaP cells. PMID- 11911848 TI - Involvement of phosphatidylinositol-3-kinase in membrane ruffling induced by P glycoprotein substrates in multidrug-resistant carcinoma cells. AB - P-glycoprotein (P-gp) is a transmembrane protein that transports a variety of structurally and functionally diverse drugs. We recently found that the interaction of drugs with P-gp promoted invasion and metastasis. In this study, we sought to determine the mechanism by which the interaction of P-gp with its substrates leads to the earliest membrane changes associated with cellular invasion, i.e., membrane ruffling. We focused on the activation of phosphatidylinositol-3-kinase (PI-3-kinase), a lipid kinase that regulates actin cytoskeletal organization and cell movement. Sensitive or multidrug-resistant (MDR) MCF-7 (human breast cancer) or KB (human oral carcinoma) cells were treated with drugs or vehicle, and then were stained with phalloidin-tetramethyl rhodamine isothiocyanate. Membrane ruffles were visualized using a fluorescence microscope. PI-3-kinase activity was determined by an in vitro immune-complex kinase assay and thin-layer chromatography. Drugs transported by P-gp, vinblastine and trans-flupenthixol, increased membrane ruffling and PI-3-kinase activity in the MDR cell lines, MCF-7/AdrR and KBV-1, which overexpress P-gp. This effect was not seen with mechlorethamine, a drug that is not transported by P-gp, and was not detected in sensitive parental cell lines that do not express P gp. A similar effect was also observed in the MDR1 transfectant, MCF-7/BC-19. Wortmannin, an inhibitor of PI-3-kinase, blocked the effect of VBL and tFPT on membrane ruffling and the activity of PI-3-kinase in MDR cells. These results indicate that drugs transported by P-gp induce membrane ruffling, an early indicator of cellular motility and metastatic potential, in cancer cells overexpressing P-gp and that this effect may be mediated through activation of PI 3-kinase. PMID- 11911849 TI - Bleomycin genotoxicity and amifostine (WR-2721) cell protection in normal leukocytes vs. K562 tumoral cells. AB - Recent advances in chemotherapy have focused on the benefit of high dose regimens, increasing the dose intensity of conventional chemotherapy. However, unacceptable cytotoxicity and genotoxicity on normal cells often impairs the proper management of patients. Phosphoaminothiol WR-1065, the active metabolite of amifostine, appears to protect normal cells and tissues against cytotoxic exposure to radiation or chemotherapeutic agents. Nevertheless, there is disagreement in findings on amifostine protection against bleomycin-induced severe side effects which have suggested that amifostine effectiveness against bleomycin-induced genotoxicity in normal leukocytes and tumour line cells K562 be studied. DNA damage was detected by single cell gel electrophoresis (or Comet) assay, a technique able to detect DNA strand breaks, alkali-labile sites and incomplete excision repair events in individual cells and which appears to be an ideal tool for assessing variability in response of different cell types in vitro. WR-2721 appears to selectively protect healthy leukocytes but not K562 tumoral cells. On the other hand, data on the inter- and intra-individual sensitivity to bleomycin and amifostine suggest that individual metabolic/genetic differences and other factors relating to lifestyle may be responsible for response variability. Application of the Comet assay in appropriate clinical settings to test the sensitivity of patients when undergoing chemotherapy appears possible. PMID- 11911850 TI - Stimulation of catecholamine biosynthesis via the protein kinase C pathway by endothelin-1 in PC12 rat pheochromocytoma cells. AB - It has been reported that endothelins (ETs) stimulate catecholamine release from chromaffin cells. However, it is not known whether ETs also affect catecholamine biosynthesis. Thus, using a rat pheochromocytoma cell line, PC12, we examined the effects of ETs on catecholamine biosynthesis. The mRNA level and activity of tyrosine hydroxylase (TH), a rate-limiting enzyme in catecholamine biosynthesis, were increased significantly by endothelin-1 (ET-1) (100nM). These stimulatory effects were inhibited completely by a blocker for the A-type endothelin receptor, BQ-123 [cyclo(D-alpha-aspartyl-L-prolyl-D-valyl-L-leucyl-D-tryptophyl)] (1 microM), but not by a blocker for the B-type endothelin receptor, BQ-788 (N cis 2,6-dimethylpiperidinocarbonyl-L-gamma-methylleucyl-D-1 methoxycarbonyltryptophanyl-D-norleucine (1 microM). Also, Ro-32-0432 (3-[8 [(dimethylamino)methyl]-6,7,8,9-tetrahydropyrido-[1,2-a]indol-10-yl]-4-(1-methyl 3-indolyl)-H-pyrrole-2,5-dione hydrochloride) (100nM), a protein kinase C inhibitor, completely inhibited ET-1-induced increases in TH activity and mRNA level. Furthermore, ET-1 (100nM) significantly stimulated protein kinase C activity, as well as inositol 1,4,5-triphosphate production; these stimulatory effects were abolished by BQ-123 but not by BQ-788. Moreover, ET-1 (100nM) significantly increased both the TH-protein level and the intracellular catecholamine content. By contrast to ET-1, endothelin-3 did not affect catecholamine synthesis. These results indicate that ET-1, but not ET-3, stimulates catecholamine synthesis through the PKC pathway in PC12 cells. Also, the use of selective ET receptor antagonists suggests that the effects of ET-1 on catecholamine biosynthesis are mediated through ET(A). PMID- 11911851 TI - Effect of clofibrate administration on the esterification and deesterification of steroid hormones by liver and extrahepatic tissues in rats. AB - Treatment of rats with clofibrate markedly stimulated the liver microsomal esterification of estradiol, testosterone, pregnenolone, dehydroepiandrosterone, and corticosterone by acyl-CoA:steroid acyltransferase. This enzyme catalyzes the esterification of estradiol with long-chain fatty acids in both liver and extrahepatic tissues. In untreated control rats, brain had the highest acyltransferase activity per milligram of microsomal protein for estradiol esterification (3- to 4-fold higher than in the liver). Although, treatment of rats with clofibrate stimulated the esterification of estradiol by 9- to 14-fold in the liver, estradiol esterification in kidney, lung, brain, uterus, fat, and mammary glands was not increased, indicating that liver may be uniquely sensitive to induction of acyl-CoA:estradiol acyltransferase by clofibrate. In additional studies, esterase activity for hydrolysis of the oleoyl ester of estradiol was determined in control and clofibrate-treated rats. Clofibrate administration increased esterase activity by an average of 107% in fat and 70% in liver. The results indicate that treatment of rats with clofibrate stimulates the hepatic formation of highly lipophilic fatty acid esters that can be hydrolyzed in the liver and in extrahepatic tissues to the parent steroid hormone by a clofibrate inducible esterase. PMID- 11911852 TI - Exendin-4 increases insulin sensitivity via a PI-3-kinase-dependent mechanism: contrasting effects of GLP-1. AB - The insulinotropic agent, exendin-4, is a long-acting analogue of glucagon-like peptide-1 (GLP-1) which improves glucose tolerance in humans and animals with diabetes, but the underlying mechanisms and the effects of exendin-4 on peripheral (muscle/fat) insulin action are unclear. Previous in vivo and clinical studies have been difficult to interpret because of complex, simultaneous changes in insulin and glucagon levels and possible effects on hepatic metabolism. Thus, the comparative effects of exendin-4 and GLP-1 on insulin-stimulated 2 [3H]deoxyglucose (2-DOG) uptake were measured in fully differentiated L6 myotubes and 3T3-adipocytes, including co-incubation with inhibitors of the PI-3-kinase (wortmannin) and mitogen-activated protein (MAP) kinase (PD098059) pathways. In L6 myotubes, there was a concentration-dependent and PI-3-kinase-dependent increase in insulin-stimulated 2-DOG uptake with exendin-4 and GLP-1, e.g. for exendin-4 the C(I-200) value (concentration of insulin required to increase 2-DOG uptake 2-fold) decreased from 1.3 +/- 1.4 x 10(-7)M (insulin alone, n=16) to 5.9 +/- 1.3 x 10(-8)M (insulin+exendin-4 0.1nM, n=18, P<0.03). A similar insulin sensitizing effect was observed with exendin-4 in 3T3-adipocytes, but GLP-1 had no effect on adipocyte insulin sensitivity. In conclusion, this is the first direct evidence showing that exendin-4 increases insulin-stimulated glucose uptake in muscle and fat derived cells via a pathway that involves PI-3-kinase activation. Furthermore, the contrasting responses of exendin and GLP-1 in 3T3 adipocytes suggest that the peripheral insulin-sensitizing effect of exendin-4 (in contrast to the insulinotropic effect) does not involve the GLP-1 receptor pathway. PMID- 11911853 TI - Effects of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)-ethane (p,p'-DDT) on 3T3-L1 and 3T3-F442A adipocyte differentiation. AB - Based upon our initial observations that 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (p,p'-DDT) induces a concentration-dependent increase in 3T3-L1 adipocyte differentiation, the mechanism of the p,p'-DDT-induced adipocyte differentiation was studied, using 3T3-L1 and 3T3-F442A cells. Since, it is known that the differentiation of the 3T3-L1 adipocyte cell line involves the induction of the transcription factors CCAAT enhancer binding protein beta (C/EBPbeta), peroxisome proliferator-activated receptor gamma (PPARgamma), and C/EBPalpha, the possible role of these factors in p,p'-DDT-induced adipocyte differentiation had to be examined. It was found that p,p'-DDT-treated 3T3-L1 cells showed a concentration dependent increase in the nuclear levels of both PPARgamma and C/EBPalpha protein. On the other hand, treatment with p,p'-DDT (20 microM) did not affect the expression pattern of C/EBPbeta protein during differentiation. Gel shift analysis of nuclear proteins for binding to the C/EBP recognition site of DNA showed an increase in binding activity at day 2 of differentiation in p,p'-DDT treated cells. Supershift analysis revealed that this rise was caused mainly by a dramatic increase in the abundance of the C/EBPalpha-DNA complex. Similar increases were observed at days 4 and 7 after the induction of differentiation. Tumor necrosis factor alpha induced a strong inhibition of adipocyte differentiation, which was reversed by co-treatment with troglitazone, an activator of PPARgamma. p,p'-DDT was unable to reverse the inhibitory effect of tumor necrosis factor alpha on adipocyte differentiation in 3T3-L1 cells. 3T3 F442A is another preadipocyte cell line that can be induced to differentiate into adipocytes in the presence of insulin and fetal bovine serum. p,p'-DDT (20 microM) induced an alteration in the morphology of these cells at day 2 after the induction of differentiation. These cells however, were unable to become fully differentiated adipocytes. These data showed, therefore, the ability of p,p'-DDT to alter the differentiation process of adipocyte cell lines through the modification of transcription factors regulating this event. PMID- 11911854 TI - Differentiation-dependent repression of c-myc, B22, COX II and COX IV genes in murine erythroleukemia (MEL) cells. AB - Murine erythroleukemia (MEL) cells provide a valuable model system for uncovering the cellular and molecular mechanisms of differentiation of proerythroid cells in culture. In order to characterize genes and gene expression patterns unique for erythropoiesis, we: (i) cloned and sequenced a 226bp cDNA encoding portion of the 3'-end B22 subunit of mitochondrial NADH-ubiquinone oxidoreductase (complex I); (ii) assessed the steady state level of RNA transcripts encoded by B22, cytochrome c oxidase (COX II, COX IV) and c-myc genes in MEL cells undergoing terminal differentiation induced by dimethylsulfoxide (DMSO) and/or 2-(3 ethylureido)-6-methylpyridine; and (iii) investigated whether the gene expression patterns of B22, COX IV and c-myc genes seen in differentiating cells are affected by N(6)-methyladenosine, an inhibitor of commitment and RNA methylation. These studies have indicated: (a) c-myc, COX II and COX IV genes exhibited biphasic expression pattern; a transient accumulation of c-myc, COX II and COX IV mRNAs was followed by a decline after 36hr incubation with DMSO and/or 2-(3 ethylureido)-6-methylpyridine, (b) B22 gene expression declined progressively in differentiated cells, (c) blockade of differentiation of MEL cells with N(6) methyladenosine failed to prevent the transient accumulation of c-myc, COX II and COX IV mRNAs, but abrogated the irreversible expression of all four genes. These findings indicated that B22, c-myc, COX II and COX IV genes are gradually repressed in terminally differentiating MEL cells presumably via different patterns of expression (gradual vs. biphasic). Overall, these results showed that erythroid maturation of MEL cells is accompanied by transcriptional inactivation (or repression) of at least three genes encoding mitochondrial enzyme subunits involved in cell respiration. PMID- 11911855 TI - Dimerumic acid as an antioxidant from the mold, Monascus anka: the inhibition mechanisms against lipid peroxidation and hemeprotein-mediated oxidation. AB - This study aimed to investigate the antioxidant mechanism of dimerumic acid isolated as the active component with a radical scavenging action from the mold Monascus anka, traditionally used for the fermentation of foods. Dimerumic acid inhibited NADPH- and iron(II)-dependent lipid peroxidation (LPO) of rat liver microsomes at 20 and 200 microM, respectively. When ferrylmyoglobin was incubated with dimerumic acid, the myoglobin was scavenged and an electron spin resonance (ESR) signal with nine peaks was observed. The spin adduct was identified as a nitroxide radical by analysis of hyperfine structure. Similar ESR signal was also detected by incubation of dimerumic acid with peroxyl radicals. Thus, it was clarified that the antioxidant action of dimerumic acid is due to one electron donation of the hydroxamic acid group in the dimerumic acid molecule toward oxidants resulting in formation of nitroxide radical. PMID- 11911856 TI - Laminar disorganisation of mitral cells in the olfactory bulb does not affect topographic targeting of primary olfactory axons. AB - Primary olfactory neurons expressing the same odorant receptor protein typically project to topographically fixed olfactory bulb sites. While cell adhesion molecules and odorant receptors have been implicated in guidance of primary olfactory axons, the postsynaptic mitral cells may also have a role in final target selection. We have examined the effect of disorganisation of the mitral cell soma layer in mutant mice heterozygous for the beta-subunit of platelet activating factor acetylhydrolase (Lis1(-/+)) on the targeting of primary olfactory axons. Lis1(-/+) mice display abnormal lamination of neurons in the olfactory bulb. Lis1(-/+) mice were crossed with the P2-IRES-tau:LacZ line of transgenic mice that selectively expresses beta-galactosidase in primary olfactory neurons expressing the P2 odorant receptor. LacZ histochemistry revealed blue-stained P2 axons that targeted topographically fixed glomeruli in these mice in a manner similar to that observed in the parent P2-IRES-tau:LacZ line. Thus, despite the aberrant organisation of postsynaptic mitral cells in Lis1(-/+) mice, primary olfactory axons continued to converge and form glomeruli at correct sites in the olfactory bulb. Next we examined whether challenging primary olfactory axons in adult Lis(-/+) mice with regeneration would affect their ability to converge and form glomeruli. Following partial chemical ablation of the olfactory neuroepithelium with dichlobenil, primary olfactory neurons die and are replaced by newly differentiating neurons that project axons to the olfactory bulb where they converge and form glomeruli. Despite the aberrant mitral cell layer in Lis(-/+) mice, primary olfactory axons continued to converge and form glomeruli during regeneration. Together these results demonstrate that the convergence of primary olfactory axons during development and regeneration is not affected by gross perturbations to the lamination of the mitral cell layer. Thus, these results support evidence from other studies indicating that mitral cells do not play a major role in the convergence and targeting of primary olfactory axons in the olfactory bulb. PMID- 11911857 TI - Different trends in modulation of NMDAR1 and NMDAR2B gene expression in cultured cortical and hippocampal neurons after lead exposure. AB - Exposure to heavy metal lead (Pb(2+)) has been reported to cause problems in cognitive functions of the brain, e.g. memory loss and difficulties in mental development. N-Methyl-D-aspartate receptors (NRs) are important molecules that are known to be involved in mediation of learning and memory. In order to investigate the effects of Pb(2+) on the gene expression of NR1 and NR2B subunits in neurons, primary cell cultures of rat cortical and hippocampal neurons were employed. After treatments with different concentrations of Pb(2+) ions in culture medium (0, 5, 10, 25 and 50 microM), the cellular localization of Pb(2+) in neurons was evaluated by laser scan confocal microscopy by using a Pb(2+) ion specific fluorescence probe. In addition, the gene expression of NR1 and NR2B subunits was determined by reverse transcriptase-polymerase chain reaction, immunofluorescence and Western blotting. The results of the present study showed that both cortical and hippocampal neurons accumulated intracellular Pb(2+) in accordance with the concentrations of Pb(2+) ions present in the culture medium. After Pb(2+) treatments, levels of NR1 mRNA, immunoreactivity and protein were found to be unchanged but levels of NR2B mRNA, immunoreactivity and protein were found to be significantly increased in cortical neurons. In contrast, both NR1 and NR2B mRNAs, immunoreactivity and proteins were found to be significantly decreased in hippocampal neurons. The changes in gene expression were found to be dose dependent in accordance with the Pb(2+) concentrations. The present results indicate that Pb(2+) has a differential effect on the expression of NR1 and NR2B subunits in cortical and hippocampal neurons, respectively. It is likely that the toxic effects of Pb(2+) may cause differential damage to different types of memory that are mediated by cortical and hippocampal neurons, respectively. PMID- 11911858 TI - Evidence for endogenous agmatine in hypothalamo-neurohypophysial tract and its modulation on vasopressin release and Ca2+ channels. AB - Agmatine, decarboxylated from arginine by arginine decarboxylase, is particularly prominent in the hypothalamus. The present study utilized the rat hypothalamo neurohypophysial system to determine expression and "pre-synaptic" modulation of agmatine in the central nervous system (CNS). Under confocal-laser scanning, agmatine-like immunoreactivity (Agm-LI) was found enriched in arginine vasopressin (AVP)-containing magnocellular neurons of the supraoptic nuclei (SON) and paraventricular nuclei (PVN). In addition, using electron microscopy, Agm-LI was found closely associated with large neurosecretory-like vesicles in neurohypophysial nerve terminals of posterior pituitary gland. Radioimmunoassay revealed that 10 and 30 microM agmatine concentration-dependently inhibited the depolarization-evoked AVP release from isolated neurohypophysial terminals. Using perforated patch-clamp, effects of agmatine on whole-terminal voltage-gated ion currents in the isolated neurohypophysial nerve terminals were examined. While it did not significantly affect either tetrodotoxin (TTX)-sensitive Na(+) or sustained Ca(2+)-activated K(+) channel currents, agmatine (1-40 microM) inhibited Ca(2+) channel currents in approximately 53% of the total nerve terminals investigated. The onset of inhibitory effect was immediate, and the inhibition was reversible and concentration-dependent with an IC(50)=4.6 microM. In the remaining (approximately 47%) neurohypophysial nerve terminals, only a higher (120 microM) concentration of agmatine could moderately inhibit Ca(2+) channel currents. The results suggest that: (1) endogenous agmatine is co expressed in AVP-containing, hypothalamic magnocellular neurons of the SON/PVN and in neurohypophysial nerve terminals of posterior pituitary gland; (2) agmatine may serve as a physiological neuromodulator by regulating the voltage gated Ca(2+) channel and, as a result, the release of AVP from neurohypophysial nerve terminals. PMID- 11911859 TI - Intrapreoptic microinjection of TNF-alpha enhances non-REM sleep in rats. AB - Tumor necrosis factor-alpha (TNFalpha) is involved in sleep regulation. Peripheral or central administration of TNFalpha induces non-rapid eye movement sleep (NREMS) in many species. However, the brain site responsible for TNF enhanced NREMS remains unclear. Thus, we tested the hypothesis that the preoptic area (POA) of the anterior hypothalamus, a crucial site for sleep regulation, is involved in TNF-induced sleep responses in rats. Unilateral microinjection of TNFalpha (2, 20 and 100 ng) or a TNF receptor fragment (TNFRF; 1.25, 5.0 and 12.5 microg) into the POA was performed at dark onset and light onset, respectively. The two higher doses of TNFalpha increased NREMS and brain temperature with little effect on REMS and EEG slow wave activity. These effects were lost after the heat-treatment of TNFalpha. The two higher doses of the TNFRF decreased NREMS without affecting the other parameters measured. Combined with previous results showing diurnal variations of TNFalpha in the hypothalamus, the present data suggest that POA TNFalpha is involved, in part, in the regulation of physiological sleep. PMID- 11911860 TI - Neuroprotective effects of the sodium channel blocker RS100642 and attenuation of ischemia-induced brain seizures in the rat. AB - Seizurogenic activity develops in many patients following brain injury and may be involved in the pathophysiological effects of brain trauma and stroke. We have evaluated the effects of the use-dependent sodium channel blocker RS100642, an analog of mexiletine, as a neuroprotectant and anti-seizure agent in a rat model of transient middle cerebral artery occlusion (MCAo). Post-injury treatment with RS100642 (0.01-5.0 mg/kg) dose-dependently reduced brain infarction, improved functional recovery of electroencephalographic (EEG) power, and improved neurological outcome following 2 h of MCAo and 24 h recovery. This effect was more potent and offered a larger reduction of brain infarct volume than a maximal neuroprotective dose of mexiletine (10.0 mg/kg). Furthermore, brain seizure activity recorded following 1 h MCAo and 72 h of recovery in injured rats was either completely blocked (30 min pre-MCAo treatment) or significantly reduced (30 min post-MCAo treatment) with RS100642 (1.0 mg/kg) treatment resulting in greater than 60% reduction of core brain infarct. These results indicate that brain seizure activity during MCAo likely contributes to the pathophysiology of brain injury and that RS100642 may be an effective neuroprotective treatment not only to decrease brain injury but also to reduce the pathological EEG associated with focal ischemia. PMID- 11911861 TI - Persistence of the neuromodulatory effects of adenosine on synaptic transmission after long-term potentiation and long-term depression. AB - Adenosine modulates long-term potentiation (LTP) and long-term depression (LTD) in the hippocampus. We tested whether induction of LTP or LTD might reciprocally modify the role of adenosine as an inhibitory modulator of synaptic transmission in the hippocampus. The experiments were performed on hippocampal slices of the rat. Two separate sets of the Schaffer pathway were alternately stimulated. Evoked field excitatory postsynaptic potentials (fEPSPs) were recorded extracellularly from CA1 stratum radiatum. Long-term potentiation (LTP) was induced by high-frequency stimulation and long-term depression (LTD) by low frequency stimulation. The inhibitory effects of the adenosine analogue, 2 chloroadenosine (CADO, 0.1-5 microM), on the fEPSP slope were similar in both pathways (EC(50)=0.72 (95% confidence intervals: 0.50-1.1) microM and EC(50)=0.84 (0.55-1.3) microM, n=6). After induction of LTP in the test pathway, a second concentration-response curve was obtained. CADO was significantly less potent as compared to the first concentration-response curve, however the inhibitory effects of CADO were still similar in the potentiated pathway (EC(50)=2.2 (1.6 3.1) microM) and in the control pathway (EC(50)=2.1 (1.5-3.0) microM, n=6). The inhibitory effects of CADO (0.1-5 microM) were also not significantly different in the pathway where LTD was previously induced (EC(50)=1.7 (1.5-2.0) microM), compared to the control non-depressed pathway (EC(50)=1.7 (1.4-2.0) microM, n=6). In conclusion, the neuromodulatory action of adenosine seems to be maintained in the presence of substantial variations in long-term synaptic efficiency during LTP or LTD. PMID- 11911862 TI - Repeated generalized audiogenic seizures induce plastic changes on acoustically evoked neuronal firing in the amygdala. AB - Repetition of audiogenic seizures (AGS) (AGS kindling) results in increases in the duration of convulsive behavior and the emergence of cortical epileptiform EEG activity. These changes involve expansion of the neuronal network subserving these seizures. The amygdala (AMG) is postulated to become involved in this expanded network, but the neurophysiological basis of this process is unknown. The present study examined changes in chronically-recorded extracellular neuronal firing patterns in the lateral nucleus of AMG (LAMG) induced by AGS kindling in behaving genetically epilepsy-prone rats (GEPR-9s). Before AGS kindling, onset only (36.1%), onset-delayed (50%) and delayed-only (13.9%) patterns of response to acoustic stimuli were observed. Neuronal firing was greatly suppressed following systemically administered uncompetitive NMDA antagonist (ketamine, 30 mg/kg, i.p.) with complete recovery by 4 h. After AGS kindling, LAMG neurons displayed a significantly increased incidence of onset-only patterns (93.3%, at 0.5 Hz), and mean acoustic responsiveness was also significantly increased (516.2% of control). LAMG neurons fired tonically during tonic convulsions and exhibited burst firing during post-tonic clonus. Greater acoustically-induced synchronization of LAMG firing, as indicated by elevated responsiveness and increased concentration of firing near the stimulus onset, may be critical for mediating the behavioral and EEG changes induced by AGS kindling. LAMG neuronal firing increases induced by AGS kindling may initiate these pathophysiological alterations, in part, by enhanced glutamate receptor-mediated excitation. This possibility is supported by the previously observed ability of an NMDA antagonist to reverse AGS kindling when focally microinjected into AMG, and the blockade of LAMG firing by administration of an uncompetitive NMDA antagonist observed in the present study. PMID- 11911863 TI - L-carnitine accelerates the in vitro regeneration of neural network from adult murine brain cells. AB - The development, growth and regeneration of nerve cells remain an unresolved issue. The up-to-date reported brain repair mechanisms are numerous and evidence suggests that, apart from the required trophism, tropism, microenvironment and specificity of the brain, a plethora of chemical, physiological and immunological compounds can contribute to such events. Among these compounds, we concentrated our interest on L-carnitine (L-Cn), which regulates the beta-oxidation of long chain fatty acids necessary for brain development, myelinization and growth. In contrast to fetal brain cells that grow easily in culture, adult brain cells show limited neurogenesis. Here, using adult brain cells from experimental mice, we show that although L-Cn does not improve their proliferative activity in short term cultures, it accelerates the growth and differentiation of neurons, astrocytes, oligodendrocytes and ependymal cells from neurospheres in long-term cultures. Thus, the formation of a confluent neural network requires a 2-month period in culture. These observations provide new insights for in vivo use of L Cn to support brain cell development in cases of injury or brain degenerative diseases. PMID- 11911864 TI - Control of serotonergic neurons in the dorsal raphe nucleus by the lateral hypothalamus. AB - Anatomical evidence indicates the presence of projections from the lateral hypothalamus to serotonergic (5-hydroxytryptamine, 5-HT) neurons of the dorsal raphe nucleus (DR). Using dual probe microdialysis and extracellular recordings in the DR, we show that the application of GABAergic agents in the lateral hypothalamus modulates the activity of 5-HT neurons in the DR. GABA and bicuculline or baclofen, applied in the lateral hypothalamus significantly reduced and increased, respectively, the 5-HT output in the DR. Likewise, the intrahypothalamic application of GABA and bicuculline reduced (14/20 neurons) and increased (8/12 neurons), respectively, the firing rate of 5-HT neurons in the DR. A smaller percentage of neurons, however, were excited by GABA (3/20) and inhibited by bicuculline (1/12). Application of tetrodotoxin in the lateral hypothalamus suppressed the local 5-HT output and reduced that in the DR. The 5 HT output in the DR increased transiently soon after darkness. The hypothalamic application of GABA attenuated and that of bicuculline potentiated this spontaneous change with an efficacy similar to that seen in light conditions. These results indicate that the lateral hypothalamus is involved in the control of 5-HT activity in the DR, possibly through excitatory (major) and inhibitory (minor) inputs. PMID- 11911865 TI - Physiologically identified 5-HT2-like receptors at the crayfish neuromuscular junction. AB - The model synaptic preparation of the crayfish opener neuromuscular junction is known to be responsive to exogenous application of 5-HT. The primary effect of 5 HT is an enhancement of vesicular release from the presynaptic motor nerve terminal. 5-HT is known to act through an IP(3) cascade which suggests the presence of a 5-HT(2) receptor subtype; however, this is based on vertebrate 5-HT receptor classification. We examined this possibility by using a selective agonist and two antagonists of the vertebrate 5-HT(2) receptor subtypes. The antagonist ketanserin and spiperone reduce the responsiveness of 5-HT in a dose dependent manner. The broad 5-HT(2) receptor agonist, alpha-methyl-5 hydroxytryptamine (alpha-Me-5-HT) enhances synaptic transmission, in a concentration-dependent manner, but it is not as potent as 5-HT. These results support the notion that a 5-HT(2) receptor subtype is present presynaptically on the crayfish motor nerve terminals. By knowing the types of 5-HT receptors present on the presynaptic motor nerve terminals in this model synaptic preparation, a better understanding of the mechanisms of action of 5-HT on vesicular release will be forthcoming. PMID- 11911866 TI - Effect of NXY-059 on secondary mitochondrial dysfunction after transient focal ischemia; comparison with cyclosporin A. AB - The free radical trapping agents NXY-059 and alpha-phenyl-N-tert.-butylnitrone (PBN) markedly reduce infarct volume, even when given 1 or 3 h after the start of recirculation, following 2 h of middle cerebral artery (MCA) occlusion in rats. Their anti-ischemic effects are shared by the two immunosuppressants cyclosporin A (CsA) and FK506. Interestingly, CsA causes an additional reduction in infarct volume when given after only 5 min of recirculation, possibly reflecting blockade of a mitochondrial permeability transition (MPT) pore. PBN, CsA and FK506 are known to ameliorate the secondary dysfunction of mitochondrial function, as assessed in vitro, which occurs during the first 4-6 h of recirculation. The present experiments were undertaken to assess whether NXY-059 reduces tissue damage by acting directly on mitochondrial membranes, and provided that this is the case, if blockade of an MPT is involved. The results were compared to those of CsA, which thus served as a reference compound. NXY-059 was given i.v. after 5 min and 1 h, and CsA after 5 min of recirculation. Both NXY-059 and CsA reduced infarct volumes to about 30% of control, prevented the secondary decline in mitochondrial respiratory function during recirculation, and reduced the mitochondrial release of cytochrome c after 6 and 24 h of recirculation. However, NXY-059 failed to block the effect of Ca(2+) on mitochondrial swelling in vitro, as CsA did. Furthermore, NXY-059, given after 5 min of recirculation, did not reproduce the effects of CsA. The results thus suggest that NXY-059 exerts its effects on mitochondria by indirect mechanisms. PMID- 11911867 TI - The use of flow cytometry to evaluate temporal changes in inflammatory cells following focal cerebral ischemia in mice. AB - Recent studies indicate that inflammation following cerebral ischemia contributes to neuronal damage. The local activation of resident cells and efficient recruitment of leukocytes into the central nervous system are critical steps in this inflammatory process. Here we describe studies using flow cytometry to examine the temporal pattern of inflammatory cell activation and infiltration following transient middle cerebral artery occlusion (MCAO) in mice. We found an increase in activated microglia/macrophages as early as 18 h post occlusion, which peaked at 48 h and remained abundant at 96 h post occlusion. Neutrophils were significantly increased by 48 h and remained elevated at 96 h post occlusion. T lymphocytes were increased relatively late (72 and 96 h) post occlusion. The flow cytometry data correlate well both quantitatively and qualitatively with immunohistochemistry analysis performed on the same mice. The present study demonstrates the power of flow cytometry in analyzing the inflammatory process following cerebral ischemia and offers temporal information on the cellular changes in mice following transient MCAO. PMID- 11911868 TI - Peripartum interneuronal coupling in the supraoptic nucleus. AB - In the supraoptic nucleus (SON), the incidence of conducting gap junctions (gjs), as indicated by dye coupling, is low in cycling females, but dramatically elevated in nursing mothers. Functionally, this is consistent with the well established presence of synchronous milk ejection bursts among oxytocin neurons only in the lactating rat. In situ hybridization data, however, revealed elevated gj mRNA expression on the last day of pregnancy, a time when burst firing by putative oxytocin neurons is absent. Using Lucifer Yellow dye coupling, we determined the incidence of high conductance gjs in SONs of proestrous, immediately prepartum, postpartum non-lactating, lactating day 1, and lactating day 9-10 rats. Results indicate that coupling incidence is high only at times when milk ejection bursts are known to occur, and that the elevated gj mRNA expression seen on the last day of pregnancy does not indicate conducting gjs. It is suggested that gj conductance states, but not gj expression, are modulated by plasma estradiol titers. PMID- 11911869 TI - Possible involvement of a muscarinic receptor in the anti-allodynic action of a 5 HT2 receptor agonist in rats with nerve ligation injury. AB - Intrathecal administration of 5-HT(2) receptor agonists produces an anti allodynic effect in a rat model of neuropathic pain. Several non-serotonergic neurotransmitters have been implicated these anti-nociceptive effects. In the present study, intrathecal pre-treatment with the muscarinic receptor antagonist atropine (10 and 30 microg) and pirenzepine (10 microg) reversed the anti allodynic effect of the 5-HT(2) receptor agonist alpha-methyl-5 hydroxytryptamine, unlike various other antagonists. Thus, muscarinic receptors may be involved in the anti-allodynic action of intrathecally injected 5-HT(2) receptor agonist. PMID- 11911870 TI - GLUT8 glucose transporter is localized to excitatory and inhibitory neurons in the rat hippocampus. AB - Fluorescence immunohistochemistry was performed to characterize the distribution and phenotype of GLUT8-positive neurons in rat brain and to compare the cellular distribution of GLUT8 with GLUT3 in the hippocampus. Based upon the absence of co localization with the non-neuronal markers GFAP (astroglial) and OX42 (microglial), it appears that GLUT8 is expressed exclusively in neurons. At the cellular level, GLUT8 immunofluorescence was localized to neuronal cell bodies and the most proximal dendrites of inhibitory and excitatory neurons while GLUT3 immunofluorescence was localized to the neuropil in the hippocampus. These results demonstrate that GLUT8 is a neuron-specific glucose transporter expressed in the neuronal cell bodies of excitatory and inhibitory neurons in the rat hippocampus. PMID- 11911871 TI - Depletion of cortical allopregnanolone potentiates stress-induced increase in cortical dopamine output. AB - In freely moving rats finasteride markedly reduced the cortical content of allopregnanolone. This treatment significantly prolonged the increase in the extracellular concentration of dopamine in the prefrontal cortex induced by foot shock. Moreover, finasteride enhanced both maximal increase of dopamine and its duration elicited by a single injection of the anxiogenic drug FG 7142. These results suggest that endogenous allopregnanolone may modulate the excitatory response of cortical dopaminergic neurons to stressful and anxiogenic stimuli. PMID- 11911872 TI - Functional protein modules. The First International EURESCO Conference. Seefeld, Austria. October 2001. PMID- 11911873 TI - Interaction domains: from simple binding events to complex cellular behavior. AB - Many of the signaling pathways and regulatory systems in eukaryotic cells are controlled by proteins with multiple interaction domains that mediate specific protein-protein and protein-phospholipid interactions, and thereby determine the biological output of receptors for external and intrinsic signals. Here, we discuss the basic features of interaction domains, and suggest that rather simple binary interactions can be used in sophisticated ways to generate complex cellular responses. PMID- 11911874 TI - The ENTH domain. AB - The epsin NH2-terminal homology (ENTH) domain is a membrane interacting module composed by a superhelix of alpha-helices. It is present at the NH2-terminus of proteins that often contain consensus sequences for binding to clathrin coat components and their accessory factors, and therefore function as endocytic adaptors. ENTH domain containing proteins have additional roles in signaling and actin regulation and may have yet other actions in the nucleus. The ENTH domain is structurally similar to the VHS domain. These domains define two families of adaptor proteins which function in membrane traffic and whose interaction with membranes is regulated, in part, by phosphoinositides. PMID- 11911875 TI - VHS domain -- a longshoreman of vesicle lines. AB - The VHS (Vps-27, Hrs and STAM) domain is a 140 residue long domain present in the very NH2-terminus of at least 60 proteins. Based on their functional characteristics and on recent data on the involvement of VHS in cargo recognition in trans-Golgi, VHS domains are considered to have a general membrane targeting/cargo recognition role in vesicular trafficking. Structurally, VHS is a right-handed superhelix of eight helices with charged surface patches probably serving as sites of protein-protein recognition and docking. PMID- 11911876 TI - The Eps15 homology (EH) domain. AB - The Eps15 homology (EH) domain was originally identified as a motif present in three copies at the NH2-termini of Eps15 and of the related molecule Eps15R. Both of these molecules are substrates for the tyrosine kinase activity of the epidermal growth factor receptor and hence the name 'Eps15 homology' or EH domain [Wong et al. (1994) Oncogene 9, 1591-1597; Wong et al. (1995) Proc. Natl. Acad. Sci. USA 92, 9530-9534; Fazioli et al. (1993) Mol. Cell. Biol. 13, 5814-5828] was derived. The motif was subsequently found in several proteins from yeast to nematode, thus establishing its evolutionary conservation. Initial studies with filter-binding assays and phage-displayed libraries demonstrated its protein:protein interaction abilities and identified specific ligands. Subsequently, structural analyses established the molecular bases of recognition between EH domains and cognate peptides. To date, several EH-containing and EH binding proteins have been identified, which establish in the cell a network of protein:protein interactions, defined as the EH network. This network coordinates cellular functions connected with endocytosis, actin remodeling and intracellular transduction of signals. PMID- 11911877 TI - WW and SH3 domains, two different scaffolds to recognize proline-rich ligands. AB - WW domains are small protein modules composed of approximately 40 amino acids. These domains fold as a stable, triple stranded beta-sheet and recognize proline containing ligands. WW domains are found in many different signaling and structural proteins, often localized in the cytoplasm as well as in the cell nucleus. Based on analyses of seven structures of WW domains, we discuss their diverse binding preferences and sequence conservation patterns. While modeling WW domains for which structures have not been determined we uncovered a case of potential molecular and functional convergence between WW and SH3 domains. The binding surface of the modeled WW domain of Npw38 protein shows a remarkable similarity to the SH3 domain of Sem5 protein, confirming biochemical data on similar binding predilections of both domains. PMID- 11911878 TI - Can we infer peptide recognition specificity mediated by SH3 domains? AB - Protein interaction domain families that modulate the formation of macromolecular complexes recognize specific sequence or structural motifs. For instance SH3 and WW domains bind to polyproline peptides while SH2 and FHA domains bind to peptides phosphorylated in Tyr and Thr respectively. Within each family, variations in the chemical characteristics of the domain binding pocket modulate a finer peptide recognition specificity and, as a consequence, determine the selection of functional protein partners in vivo. In the proteomic era there is the need for reliable inference methods to help restricting the sequence space of the putative targets to be confirmed experimentally by more laborious experimental approaches. Here we will review the published data about the peptide recognition specificity of the SH3 domain family and we will propose a classification of SH3 domains into eight classes. Finally, we will discuss whether the available information is sufficient to infer the recognition specificity of any uncharacterized SH3 domain. PMID- 11911879 TI - EVH1 domains: structure, function and interactions. AB - Drosophila enabled/vasodilator-stimulated phosphoprotein homology 1 (EVH1) domains are 115 residue protein-protein interaction modules which provide essential links for their host proteins to various signal transduction pathways. Many EVH1-containing proteins are associated closely with actin-based structures and are involved in re-organization of the actin cytoskeleton. EVH1 domains are also present in proteins enriched in neuronal tissue, thus implicating them as potential mediators of synaptic plasticity, linking them to memory formation and learning. Like Src homology 3, WW and GYF domains and profilin, EVH1 domains recognize and bind specific proline-rich sequences (PRSs). The binding is of low affinity, but tightly regulated by the high specificity encoded into residues in the protein:peptide interface. In general, a small (3-6 residue) 'core' PRS in the target protein binds a 'recognition pocket' on the domain surface. Further affinity- and specificity-increasing interactions are then formed between additional domain epitopes and peptide 'core-flanking' residues. The three dimensional structures of EVH1:peptide complexes now reveal, in great detail, some of the most important features of these interactions and allow us to better understand the origins of specificity, ligand orientation and sequence degeneracy of target peptides, in low affinity signalling complexes. PMID- 11911880 TI - How do 14-3-3 proteins work?-- Gatekeeper phosphorylation and the molecular anvil hypothesis. AB - 14-3-3 proteins were the first signaling molecules to be identified as discrete phosphoserine/threonine binding modules. This family of proteins, which includes seven isotypes in human cells and up to 15 in plants, plays critical roles in cell signaling events that control progress through the cell cycle, transcriptional alterations in response to environmental cues, and programmed cell death. Despite over 30 years of research, distinct roles for most isotypes remain unknown. Though 14-3-3 proteins perform different functions for different ligands, general mechanisms of 14-3-3 action include changes in activity of bound ligands, altered association of bound ligands with other cellular components, and changes in intracellular localization of 14-3-3-bound cargo. We present a speculative model where binding of 14-3-3 to multiple sites on some ligands results in global ligand conformational changes that mediate their biological effects. For these multi-site ligands, one binding site is likely to function as a 'gatekeeper' whose phosphorylation is necessary for 14-3-3 binding but may not always be sufficient for full biological activity. If correct, then 14-3-3 may prove to be a bona fide phosphodependent signaling chaperone. PMID- 11911881 TI - The FHA domain. AB - The forkhead-associated (FHA) domain is a small protein module recently shown to recognize phosphothreonine epitopes on proteins. It is present in a diverse range of proteins in eukaryotic cells, such as kinases, phosphatases, kinesins, transcription factors, RNA-binding proteins, and metabolic enzymes. It is also found in a number of bacterial proteins. This suggests that FHA domain-mediated phospho-dependent assembly of protein complexes is an ancient and widespread regulatory mechanism. PMID- 11911882 TI - PTB or not PTB -- that is the question. AB - Phosphotyrosine binding (PTB) domains are structurally conserved modules found in proteins involved in numerous biological processes including signaling through cell-surface receptors and protein trafficking. While their original discovery is attributed to the recognition of phosphotyrosine in the context of NPXpY sequences -- a function distinct from that of the classical src homology 2 (SH2) domain -- recent studies show that these protein modules have much broader ligand binding specificities. These studies highlight the functional diversity of the PTB domain family as generalized protein interaction domains, and reinforce the concept that evolutionary changes of structural elements around the ligand binding site on a conserved structural core may endow these protein modules with the structural plasticity necessary for functional versatility. PMID- 11911883 TI - Pleckstrin homology domains and the cytoskeleton. AB - Pleckstrin homology (PH) domains are 100-120 amino acid protein modules best known for their ability to bind phosphoinositides. All possess an identical core beta-sandwich fold and display marked electrostatic sidedness. The binding site for phosphoinositides lies in the center of the positively charged face. In some cases this binding site is well defined, allowing highly specific and strong ligand binding. In several of these cases the PH domains specifically recognize 3 phosphorylated phosphoinositides, allowing them to drive membrane recruitment in response to phosphatidylinositol 3-kinase activation. Examples of these PH domain containing proteins include certain Dbl family guanine nucleotide exchange factors, protein kinase B, PhdA, and pleckstrin-2. PH domain-mediated membrane recruitment of these proteins contributes to regulated actin assembly and cell polarization. Many other PH domain-containing cytoskeletal proteins, such as spectrin, have PH domains that bind weakly, and to all phosphoinositides. In these cases, the individual phosphoinositide interactions may not be sufficient for membrane association, but appear to require self-assembly of their host protein and/or cooperation with other anchoring motifs within the same molecule to drive membrane attachment. PMID- 11911884 TI - The phosphatidylinositol 3-phosphate-binding FYVE finger. AB - The FYVE zinc finger domain is conserved from yeast (five proteins) to man (27 proteins). It functions in the membrane recruitment of cytosolic proteins by binding to phosphatidylinositol 3-phosphate (PI3P), which is found mainly on endosomes. Here we review recent work that sheds light on the targeting of FYVE finger proteins to PI3P-containing membranes, and how these proteins serve to regulate multiple cellular functions. PMID- 11911885 TI - Signaling to the Rho GTPases: networking with the DH domain. AB - The Dbl homology (DH) domain was first identified in the Dbl oncogene product as the limit region required for mediating guanine nucleotide exchange on the Rho family GTPase Cdc42. Since the initial biochemical characterization of the DH domain, this conserved motif has been identified in a large family of proteins. In each case, a pleckstrin homology (PH) domain immediately follows the DH domain and this tandem DH-PH module is the signature motif of the Dbl family of guanine nucleotide exchange factors (GEFs). Recent structural studies have provided significant insight into the molecular basis of guanine nucleotide exchange by Dbl family GEFs, opening the door for understanding the specificity of the DH/GTPase interaction as well as providing a starting point for understanding how the exchange activity of these proteins is modulated to achieve specific biological outcomes in the cell. PMID- 11911886 TI - WH2 domain: a small, versatile adapter for actin monomers. AB - The actin cytoskeleton plays a central role in many cell biological processes. The structure and dynamics of the actin cytoskeleton are regulated by numerous actin-binding proteins that usually contain one of the few known actin-binding motifs. WH2 domain (WASP homology domain-2) is a approximately 35 residue actin monomer-binding motif, that is found in many different regulators of the actin cytoskeleton, including the beta-thymosins, ciboulot, WASP (Wiskott Aldrich syndrome protein), verprolin/WIP (WASP-interacting protein), Srv2/CAP (adenylyl cyclase-associated protein) and several uncharacterized proteins. The most highly conserved residues in the WH2 domain are important in beta-thymosin's interactions with actin monomers, suggesting that all WH2 domains may interact with actin monomers through similar interfaces. Our sequence database searches did not reveal any WH2 domain-containing proteins in plants. However, we found three classes of these proteins: WASP, Srv2/CAP and verprolin/WIP in yeast and animals. This suggests that the WH2 domain is an ancient actin monomer-binding motif that existed before the divergence of fungal and animal lineages. PMID- 11911887 TI - Functional plasticity of CH domains. AB - With the refinement of algorithms for the identification of distinct motifs from sequence databases, especially those using secondary structure predictions, new protein modules have been determined in recent years. Calponin homology (CH) domains were identified in a variety of proteins ranging from actin cross-linking to signaling and have been proposed to function either as autonomous actin binding motifs or serve a regulatory function. Despite the overall structural conservation of the unique CH domain fold, the individual modules display a quite striking functional variability. Analysis of the actopaxin/parvin protein family suggests the existence of novel (type 4 and type 5) CH domain families which require special attention, as they appear to be a good example for how CH domains may function as scaffolds for other functional motifs of different properties. PMID- 11911888 TI - Calmodulin signaling via the IQ motif. AB - The IQ motif is widely distributed in both myosins and non-myosins and is quite common in the database that includes more than 900 Pfam entries. An examination of IQ motif-containing proteins that are known to bind calmodulin (CaM) indicates a wide diversity of biological functions that parallel the Ca2+-dependent targets. These proteins include a variety of neuronal growth proteins, myosins, voltage-operated channels, phosphatases, Ras exchange proteins, sperm surface proteins, a Ras Gap-like protein, spindle-associated proteins and several proteins in plants. The IQ motif occurs in some proteins with Ca2+-dependent CaM interaction where it may promote Ca2+-independent retention of CaM. The action of the IQ motif may result in complex signaling as observed for myosins and the L type Ca2+ channels and is highly localized as required for sites of neuronal polarized growth and plasticity, fertilization, mitosis and cytoskeletal organization. The IQ motif associated with the unconventional myosins also promotes Ca2+ regulation of the vectorial movement of cellular constituents to these sites. Additional regulatory roles for this versatile motif seem likely. PMID- 11911889 TI - The paxillin LD motifs. AB - Adapter/scaffold proteins, through their multidomain structure, perform a fundamental role in facilitating signal transduction within cells. Paxillin is a focal adhesion adapter protein implicated in growth factor- as well as integrin mediated signaling pathways. The amino-terminus of paxillin contains five leucine rich sequences termed LD motifs. These paxillin LD motifs are highly conserved between species as well as within the paxillin superfamily. They mediate interactions with several structural and regulatory proteins important for coordinating changes in the actin cytoskeleton associated with cell motility and cell adhesion as well as in the regulation of gene expression. PMID- 11911890 TI - The spectrin repeat: a structural platform for cytoskeletal protein assemblies. AB - Spectrin repeats are three-helix bundle structures which occur in a large number of diverse proteins, either as single copies or in tandem arrangements of multiple repeats. They can serve structural purposes, by coordination of cytoskeletal interactions with high spatial precision, as well as a 'switchboard' for interactions with multiple proteins with a more regulatory role. We describe the structure of the alpha-actinin spectrin repeats as a prototypical example, their assembly in a defined antiparallel dimer, and the interactions of spectrin repeats with multiple other proteins. The alpha-actinin rod domain shares several features common to other spectrin repeats. (1) The rod domain forms a rigid connection between two actin-binding domains positioned at the two ends of the alpha-actinin dimer. The exact distance and rigidity are important, for example, for organizing the muscle Z-line and maintaining its architecture during muscle contraction. (2) The spectrin repeats of alpha-actinin have evolved to make tight antiparallel homodimer contacts. (3) The spectrin repeats are important interaction sites for multiple structural and signalling proteins. The interactions of spectrin repeats are, however, diverse and defy any simple classification of their preferred interaction sites, which is possible for other domains (e.g. src-homology domains 3 or 2). Nevertheless, the binding properties of the repeats perform important roles in the biology of the proteins where they are found, and lead to the assembly of complex, multiprotein structures involved both in cytoskeletal architecture as well as in forming large signal transduction complexes. PMID- 11911891 TI - Bromodomain: an acetyl-lysine binding domain. AB - Bromodomains, an extensive family of evolutionarily conserved protein modules originally found in proteins associated with chromatin and in nearly all nuclear histone acetyltransferases, have been recently discovered to function as acetyl lysine binding domains. More recent structural studies of bromodomain/peptide ligand complexes have enriched our understanding of differences in ligand selectivity of bromodomains. These new findings demonstrate that bromodomain/acetyl-lysine recognition can serve as a pivotal mechanism for regulating protein-protein interactions in numerous cellular processes including chromatin remodeling and transcriptional activation, and reinforce the concept that functional diversity of a conserved protein modular structure is achieved by evolutionary changes of amino acid sequences in the ligand binding site. PMID- 11911892 TI - Protein domain analysis in the era of complete genomes. AB - Domains present one of the most useful levels at which to understand protein function, and domain family-based analysis has had a profound impact on the study of individual proteins. Protein domain discovery has been progressing steadily over the past 30 years. What are the realistically achievable goals of sequence based domain analysis, and how far off are they for the sequences encoded in eukaryotic genomes? Here we address some of the issues involved in better coverage of sequence-based domain annotation, and the integration of these results within the wider context of genomes, structures and function. PMID- 11911893 TI - MINT: a Molecular INTeraction database. AB - Protein interaction databases represent unique tools to store, in a computer readable form, the protein interaction information disseminated in the scientific literature. Well organized and easily accessible databases permit the easy retrieval and analysis of large interaction data sets. Here we present MINT, a database (http://cbm.bio.uniroma2.it/mint/index.html) designed to store data on functional interactions between proteins. Beyond cataloguing binary complexes, MINT was conceived to store other types of functional interactions, including enzymatic modifications of one of the partners. Release 1.0 of MINT focuses on experimentally verified protein-protein interactions. Both direct and indirect relationships are considered. Furthermore, MINT aims at being exhaustive in the description of the interaction and, whenever available, information about kinetic and binding constants and about the domains participating in the interaction is included in the entry. MINT consists of entries extracted from the scientific literature by expert curators assisted by 'MINT Assistant', a software that targets abstracts containing interaction information and presents them to the curator in a user-friendly format. The interaction data can be easily extracted and viewed graphically through 'MINT Viewer'. Presently MINT contains 4568 interactions, 782 of which are indirect or genetic interactions. PMID- 11911894 TI - Normalization of nomenclature for peptide motifs as ligands of modular protein domains. AB - We propose a normalization of symbols and terms used to describe, accurately and succinctly, the detailed interactions between amino acid residues of pairs of interacting proteins at protein:protein (or protein:peptide) interfaces. Our aim is to unify several diverse descriptions currently in use in order to facilitate communication in the rapidly progressing field of signaling by protein domains. In order for the nomenclature to be convenient and widely used, we also suggest a parallel set of symbols restricted to the ASCII format allowing accurate parsing of the nomenclature to a computer-readable form. This proposal will be reviewed in the future and will therefore be open for the inclusion of new rules, modifications and changes. PMID- 11911895 TI - Rheological characteristics of microbial suspensions of Pseudomonas aeruginosa and Bacillus cereus. AB - The rheological properties of the bacteria Pseudomonas aeruginosa and Bacillus cereus have been investigated. The apparent viscosity of the bacterial suspensions has been measured at different conditions. The results showed that the bacterial suspensions' apparent viscosity increased with increasing biomass concentration of each of these strains. The P. aeruginosa suspension followed shear thinning behavior while B. cereus suspension followed shear thickening behavior. The shear stress versus shear rate experimental data were best represented by the Herschel-Bulkley model. The apparent viscosity of the P. aeruginosa and B. cereus suspensions decreased with increasing temperature. The relationship between the apparent viscosity and the shearing time highlighted the rheopectic behavior of the suspensions used in this work. PMID- 11911896 TI - Immobilized gellan sulfate surface for cell adhesion and multiplication: development of cell-hybrid biomaterials using self-produced fibronectin. AB - A new concept for cell-hybrid biomaterial is proposed in which human unbilical vein endothelial cells (HUVEC) are adhered to an immobilized gellan sulfate (GS) surface. Extra domain A containing fibronectin (EDA(+)FN) released from HUVEC is necessary for cell adhesion and multiplication. The material design in this study is based on these self-released cell adhesion proteins. The interaction between GS and EDA(+)FN was evaluated using the affinity constant (KA); the value obtained was 1.03x10(8) (M(-1)). These results suggest that the adhesion of HUVEC to GS may be supported by the adhesion of EDA(+)FN to GS. We also found that this new material adheres to HUVEC, allowing the reintroduction of EDA(+)FN, which is self-produced by the cell. This material is relatively easy to produce, not requiring the usual coating of adhesion proteins in pretreatment. PMID- 11911897 TI - Immobilization of glucose oxidase in polypyrrole/polytetrahydrofuran graft copolymers. AB - Glucose oxidase (GOD) was immobilized in four different conducting polymer matrices, namely: polypyrrole, (PPy), poly(pyrrole-graft-polytetrahydrofuran), (1) and (3); and poly(pyrrole-graft-polystyrene/polytetrahydrofuran), (2). The kinetic parameters V(max) and K(m), and the optimum temperature were determined for both immobilized and native enzymes. The effect of electrolysis time and several supporting electrolytes, p-toluenesulfonic acid, p-toluene sulfonic acid (PTSA), sodium p-toluene sulfonate, sodium p-toluene sulfonate (NaPTS), and sodium dodecyl sulfate, sodium dodecyl sulfate (SDS), on enzyme immobilization were investigated. The high K(m) value (59.9 mM) of enzyme immobilized in PPy was decreased via immobilization in graft copolymer matrices of pyrrole. V(max), which was 2.25 mM/min for pure PPy, was found as 4.71 mM/min for compound (3). PMID- 11911898 TI - Structure and properties of silk fibroin-poly(vinyl alcohol) gel. AB - A series of porous silk fibroin-poly(vinyl alcohol) blend gels were prepared from corresponding aqueous mixtures by freeze- or air-drying, and their structure and mechanical properties were examined. The air-dried gels had higher crystallinity and much greater strengths than the freeze-dried gels, and therefore are suitable for mechanical uses. The freeze-dried gels had characteristic porous structure potentially useful as cell culture substrate. Its structure could be systematically varied by changing freezing temperature and freeze-thaw pretreatments before drying. PMID- 11911899 TI - The parallel helices of the intermediate filaments of alpha-keratin. AB - Recent Fourier transform infrared spectroscopy (FTIR) with attenuated total reflection technique (ATR) has been applied to alpha-keratin fibers (horse-hair) extended in water both at 21 and 95 degrees C. Infrared absorption bands in the Amide 1 region indicated that at extensions to 40-50% strain in water at 21 degrees C alpha-helices had completely disappeared and parallel beta-sheets were formed [Appl. Spectrosc. 55 (2001) 552]. However, when the hair fibers were extended to the same strain at 95 degrees C in water the result was the formation of anti-parallel beta-sheets. These results suggest that the relatively more stable anti-parallel beta-state [Polymer 10 (1969) 810] is only attained in extended alpha-keratin fibers at elevated temperatures and must result from major molecular rearrangement. It was concluded that the alpha-helices in the intermediate filaments (IFs) of alpha-keratin fibers must be parallel. This is in contrast to the previously accepted orientation of anti-parallel alpha-helices, based primarily on findings of X-ray diffraction studies of the structure of beta keratin in highly extended fibers [Polymer 10 (1969) 810; Keratins, IL: Thomas Springfield (1972); Nature 316 (1985) 767]. PMID- 11911900 TI - Effect of increased PHA synthase activity on polyhydroxyalkanoates biosynthesis in Synechocystis sp. PCC6803. AB - Polyhydroxyalkanoate (PHA) synthase activity in Synechocystis sp. PCC6803 was increased two-fold by introducing the PHA biosynthetic genes of Ralstonia eutropha. The resulting recombinant Synechocystis sp. PCC6803 strain was subjected to conditions that favor PHA accumulation and the effects of various carbon sources were studied. In addition, the fine structure of both wild-type and recombinant Synechocystis sp. PCC6803 was examined using freeze-fracture electron microscopy technique. The PHA granules in the recombinant Synechocystis sp. PCC6803 were localised near the thylakoid membranes. Maximum amount of PHA accumulation was obtained in the presence of acetate, where the number of granules in the recombinant cells ranged from 4 to 6 and their sizes were in the range of 70-240 nm. In comparison to wild-type Synechocystis sp. PCC6803, recombinant cells with increased PHA synthase activity showed only a marginal increase in PHA content suggesting that PHA synthase is not the rate limiting enzyme of PHA biosynthesis in Synechocystis sp. PCC6803. PMID- 11911901 TI - Monomer composition and sequence of alginates from Pseudomonas aeruginosa. AB - Alginates from four strains of Pseudomonas aeruginosa, one mucoid strain isolated from a technical water system, one strain isolated from a patient with cystic fibrosis and two mutants of this strain with a defect which affects the O acetylation of the extracellular alginate, have been isolated and analysed for monomer composition and sequence by 13C-nuclear magnetic resonance (NMR) spectroscopy. The detected contributions of different monomer triplets (triads) were compared with values expected from a statistical chain constitution based on the given monomer ratio. While a typical algal alginate presents a nearly statistical distribution of uronic acids in the polymer chain, a strong deviation from the statistical arrangement of mannuronate (M) and guluronate (G) was found in the alginate of the mucoid strains of P. aeruginosa, being most expressed for the triad MMM. This feature is partially lost in the alginate from the mutant strains, indicating that the O-acetylation is linked to a mechanism which takes influence on the chain sequence. The strong preference for MG-pairs in the parent strain of P. aeruginosa may be connected to a stronger binding of cations in the MG-vicinity. PMID- 11911902 TI - Interaction of daunomycin antibiotic with histone H(1): ultraviolet spectroscopy and equilibrium dialysis studies. AB - Using ultraviolet spectroscopy and equilibrium dialysis techniques, we have investigated the interaction of anticancer drug, daunomycin with calf thymus histone H(1) chromosomal protein in 20 mM phosphate buffer, pH 7.0, 1 mM EDTA at room temperature. The UV spectroscopy results show that daunomycin (5.0-100 microM) decreases the absorbance of histone H(1) at 210-230 nm and induces hypochromicity in the absorption spectrum of the protein. The equilibrium dialysis data show that daunomycin binds to histone H(1) and the binding process is positive cooperative with two binding sites as Scatchard plot and Hill coefficient confirm it. The results suggest that daunomycin binds to histone H(1) and changes its conformation. PMID- 11911903 TI - The viscoelastic basis for the tensile strength of elastin. AB - Purified aortic elastin displays failure behaviour characteristic of an amorphous, noncrystalizing elastomer with failure properties showing a strong dependence on viscoelastic behaviour. Tensile breaking stresses and breaking strains measured over a range of temperatures, hydration levels, and strain rates are reducible to single curves by the application of shift factors obtained from dynamic mechanical tests. The breaking stress of rubbery elastin is similar to that found in other elastomers, but glassy elastin is about an order of magnitude less strong than expected. We suggest elastin's ability to be strengthened through viscous dissipation of strain energy and crack tip blunting is limited by its fibrillar structure. PMID- 11911904 TI - Preparation and cell compatibility of acrylamide-grafted poly(3 hydroxyoctanoate). AB - Poly(3-hydroxyoctanoate) (PHO) films were treated with plasma of different discharge powers (10-50 W) and then treated with acryl amide solutions in order to prepare films with surfaces that contained different amounts of amide groups. The surfaces were characterized by contact angle measurement, attenuated total reflectance infrared spectroscopy, electron spectroscopy for chemical analysis, and scanning electron microscopy. Results from all these measurements indicated that amide groups were present on the surfaces. The amount of amide groups increased in proportion to the discharge power of the plasma. The interaction of Chinese hamster ovary cells with these grafted surfaces was investigated. The number of cells that adhered to and grew on the surfaces was highest for films grafted at 30 W of plasma discharge power, indicating that the moderate hydrophilicity was optimal for cells to adhere and grow. The present results support the suggestion that acryl amide-grafted PHO could be used as cell compatible biomedical applications. PMID- 11911905 TI - Are high-density lipoprotein and triglyceride levels important in secondary prevention: impressions from the BIP and VA-HIT trials. AB - Two major trials, the Bezafibrate Infarction Prevention Trial (BIP) and the Veterans Affairs High-Density Lipoprotein Cholesterol Intervention Trial (VA-HIT) were conducted to clarify the contribution of correcting diminished high density lipoprotein (HDL) (and lowering triglyceride, TG) levels to the risk of cardiovascular events in patients with coronary heart disease (CHD). In BIP, bezafibrate did not significantly reduce the risk of CHD. In contrast, in VA-HIT, gemfibrozil significantly reduced the risk of CHD (22% reduction in primary end point, P=0.006). These trials differ in several respects making direct comparisons difficult. For example, the placebo arm in VA-HIT had a greater prevalence of primary events than that in BIP (22 vs. 15%). The baseline mean LDL value in BIP was also higher compared to that in VA-HIT (148 vs. 112 mg/dl; 3.82 vs. 2.89 mmol/l). Other trials (e.g., AFCAPS and LIPID) showed that patients with LDL values similar to those in BIP benefited significantly from treatment with statins. Therefore, the BIP population may have been more effectively treated with a statin. In contrast, in VA-HIT the LDL level was close to those recommended in the USA and the UK for secondary prevention (100 and 115 mg/dl; 2.6 and 3.0 mmol/l, respectively). Guidelines emphasise that the LDL level is the main treatment target. However, BIP and VA-HIT suggest that correcting HDL and TG levels may be beneficial especially when the LDL level has reached the target value. We may have become too focused on LDL levels and the use of statins. PMID- 11911907 TI - Atrial fibrillation after coronary surgery: comparison between different health care systems. AB - AIMS: No studies have evaluated the influence of management strategies in different health insurance environments on atrial fibrillation (AF). This observational study compared the incidence of and treatment strategies for postoperative AF after primary coronary bypass surgery. METHODS AND RESULTS: One insurance and one public funded location was compared: University of Michigan Health Center (USA, n=272) and Tampere University Hospital (Finland, n=314). USA patients had more co-morbidities and were treated more aggressively after acute myocardial infarction. More Finns were on beta-blockers both preoperatively (93 vs. 68%, P<0.001) and postoperatively (97 vs. 66%, P<0.001). However, AF was more frequent among Finns (38 vs. 29%, P=0.037) and present on 4.6% of cases when transferred postoperatively. No USA patients had AF at time of discharge. Mean length of stay was 8.6 days at USA, and not affected by AF. The incidence of in hospital death, strokes and multiorgan failures was similar. Multivariable analysis, adjusted for site and selection biases (propensity analysis) revealed increasing age [OR=1.063 (1.042, 1.084), P<0.0001] and use of radial arteries [OR=2.175 (1.071, 4.417), P=0.032) to be independent predictors to the incidence of postoperative AF. CONCLUSIONS: We found several major differences in patient selection and treatment strategies among primary coronary bypass patients managed in the two institutions. Despite the marked practice variation, the incidence of postoperative AF was rather similar. Despite routine use of beta-blockers, AF occurred in 29-38% of patients. However, the length of stay was not particularly affected by postoperative AF. PMID- 11911909 TI - Mechanisms of ventricular fibrillation during coronary angioplasty: increased incidence for the small orifice caliber of the right coronary artery. AB - BACKGROUND: Ventricular fibrillation (VF) is not an infrequent complication of percutaneous transluminal coronary angioplasty (PTCA). However, it is not clear why there is a marked discrepancy in the higher incidence of VF during right coronary artery (RCA) approach. METHODS AND RESULTS: We analyzed in detail every case of VF occurring in 905 consecutive PTCA procedures to investigate possible mechanisms. Sixteen patients (M/F=15/1, mean age: 71 +/- 8 years) with VF during PTCA for the RCA as Group I. Those 51 patients (M/F=48/3, mean age: 70 +/- 9 years) without VF during PTCA for the RCA engagement were designated as Group II. Patients were equipped with cardiac event recorder (CardioCall, Reynolds Medical, UK) before the PTCA, and we set the time period 1 min before and after the event. The lead II was selected to check the QRS width, QTc interval, ST segment change and RR interval before and after event. A total of 905 PTCA procedures were included. There were 561 procedures for the left coronary artery and three events (0.5%) with spontaneous VF. However, there were 16 events (4.6%) of VF during 344 PTCA procedures for the right coronary artery. The incidence of VF for the right side PTCA was significantly higher than for the left side. The orifice of RCA in Group I was smaller than Group II (orifice of RCA in Group I vs. Group II - 2.7+/ 0.8 vs. 4.1+/-1.2 mm, P<0.001). Most cases (68.7%) presented with ST segment depression before the onset of VF. CONCLUSION: A small caliber of RCA and associated ST segment changes played important roles in the patients with VF during the PTCA. PMID- 11911910 TI - Trends in case-fatality in 22968 patients admitted for the first time with atrial fibrillation in Scotland, 1986-1995. AB - BACKGROUND: Although atrial fibrillation (AF) is an important cause of cardiovascular morbidity and mortality there is a paucity of data describing hospitalisation rates and case-fatality associated with this common arrhythmia. This study examines recent trends in first-ever hospitalisations for AF in Scotland. METHODS: Using the linked Scottish Morbidity Record Scheme, we identified all 22968 patients admitted to Scottish hospitals for the first time with a principal diagnosis of AF between 1986 and 1995. For each calendar year we calculated short (30-day) and medium (31 day to 2 years) case-fatality rates. Adjusting for each patient's age, sex, deprivation status, concurrent diagnoses and prior hospitalisation status, we examined whether case-fatality rates had significantly improved during this 10-year period. RESULTS: Between 1986 and 1995 the number of men hospitalised for the first time with AF increased by 926 (125%) to 1730 per annum and the number of women and by 875 (105%) to 1712 (both P<0.001). Hospitalisation rates increased from 0.31 to 0.70/1000 men and from 0.32 to 0.65/1000 women (both P<0.001). By the end of this period the proportion of men had increased from 48 to 50%. In both sexes, the median age of patients rose--in men from 66 to 68 years and in women from 74 to 75 years (both P<0.01). Despite the increasing age of patients and greater comorbidity, short-term (30 day) case-fatality declined from 4.0 to 3.1% in men (P<0.001) and 4.1 to 3.8% (P<0.01) in women. Similarly, medium-term (31-day to 2-year) case-fatality fell from 25 to 22% in men and 27 to 25% (both P<0.001) in women. Adjusting for the age, sex, extent of deprivation, secondary diagnoses and prior hospitalisation of hospitalised patients, we found that the risk of short-term case-fatality in the 1995 male and female cohort significantly declined by 21% (P<0.05) and 24% (P<0.05), respectively, in comparison to the 1986 cohort. The adjusted risk of case-fatality in the medium term also declined significantly in men by 30% (P<0.05) over this period and by 20% (P<0.05) in women relative to 1986. CONCLUSION: The number of first-ever hospitalisations for AF has increased twofold during the 10-year period 1986-1995. Although the age of patients has progressively increased during this period, short and medium case-fatality rates have declined, especially in men. This may partly reflect better treatment of AF. However, changing admission thresholds and other factors could also have led to an apparent improvement in prognosis. Nevertheless, medium-term case fatality remains substantial after a first ever admission to hospital with AF. PMID- 11911911 TI - Coronary endothelial dysfunction and myocardial cell damage in chronic stable idiopathic dilated cardiomyopathy. AB - Impairment of endothelium-dependent vasodilatation in response to acetylcholine reflects an abnormal endothelial function. Labelled indium-111 monoclonal antimyosin antibodies enable detection of myocardial cell damage. We analysed whether endothelial dysfunction correlates with myocardial antimyosin uptake in a selected group of patients with idiopathic dilated cardiomyopathy. METHODS: Twenty-two consecutive patients with chronic stable idiopathic dilated cardiomyopathy (18 males and four females) were included. The duration of heart failure symptoms was 46+/-34 months. At inclusion, the functional class of New York Heart Association was 2.1+/-0.7. Endothelial function was evaluated using intracoronary graded concentrations of acetylcholine. Vasomotor responses of the left anterior descending coronary artery were measured by quantitative coronary analysis. Myocardial uptake of antimyosin antibodies was quantified by means of a heart-to-lung ratio (HLR). RESULTS: Eighteen patients showed endothelial dysfunction (82%) and the remaining four patients showed a normal endothelial function. There were no statistically significant differences between patients with and without endothelial dysfunction in relation to clinical, echocardiographic and hemodynamic parameters. In addition, these variables did not correlate with the magnitude of the vasomotor response to acetylcholine. Eighteen patients (82%) showed abnormal antimyosin uptake; 15 of them (83%) showed endothelial dysfunction. The global mean HLR of antimyosin uptake was 1.73+/-0.24. The coronary vasomotor response to acetylcholine correlated with the intensity of uptake of antimyosin antibodies (r=-0.45, P<0.04). CONCLUSIONS: Coronary endothelial dysfunction and myocardial antimyosin uptake was found in a high percentage of patients with chronic stable idiopathic dilated cardiomyopathy. The abnormal vasomotor response seems to be related to the degree of myocardial damage. PMID- 11911912 TI - Anti-streptokinase titers and response to streptokinase treatment in Pakistani patients. AB - In order to investigate whether anti-streptokinase (anti-SK) antibody titers have any effect on response to standard dose (1.5 million units) streptokinase (SK) therapy in patients with acute myocardial infarction, anti-SK titers in plasma were determined in 97 such patients. These patients were classified as "responders" or "non-responders" on the basis of a criteria which involved resolution of chest pain in less than 90 min after the start of thrombolysis, greater than 50% reduction of ST segment elevation in two contiguous leads showing maximum elevations in a 12-lead tracing, reperfusion arrythmias and slope of increase in creatine kinase over a 90-min period after initiation of thrombolysis. Anti-SK antibody levels in plasma were determined in these patients as well as in 50 normal healthy subjects by dissolution of clot method. Mean+/ S.D. anti-SK levels among responders, non-responders and normal healthy subjects were found to be 0.21+/-0.12, 0.25+/-0.22 and 0.21+/-0.15 million units, respectively. A comparison of these mean values by one-way ANOVA revealed no statistical difference (P=0.68). However, when compared with the reported values for a Western population, these values were found to be significantly higher (P<0.03). Whereas, compared to the reported values for an Indian population, these were significantly lower (P<0.0001). 42% of patients appeared to have responded to SK treatment, while 21% were found to be non-responders. There was no association between response to SK and anti-SK titers in these patients suggesting that the standard dose of SK (1.5 million units) may be quite appropriate for the Pakistani population requiring SK treatment for the first time. PMID- 11911913 TI - Frequency in the anomalous origin of the right coronary artery with angiography in a Turkish population. AB - OBJECTIVE: The estimate frequency of anatomic variations in origin of the right coronary artery in a Turkish population. MATERIAL AND METHOD: The angiographic data of 5253 consecutive adults patients undergoing coronary angiography were analysed retrospectively for the diagnosis of anomalous origin of the right coronary artery. RESULTS: Among 5253 adults patients, five (0.09%) patients had anomalous origin of the right coronary. They had an isolated anomalous origin of the right coronary artery. The right coronary arose from the left coronary sinus of Valsalva (there was separate orificium for the right coronary artery and the left coronary artery) in two (0.03%) patients, from above the left coronary sinus of Valsalva in three (0.05%) patients. In all patients, the anomalous origin of right coronary artery from the left sinus of Valsalva and from above the left coronary sinus of Valsalva coursed between the aorta and the pulmonary artery. CONCLUSION: The anomalous origin of the right coronary artery is a rare congenital cardiac malformation. Most patients remain asymptomatic. However, there are cases of sudden cardiac death described in the literature, indicating a potentially malign course of the disease. The angiographic recognition of this vessel may be useful for physicians dealing with diagnosis and treatment of the anomaly of the right coronary artery. PMID- 11911914 TI - Prognosis and risk indicators of death during a period of 10 years for women admitted to the emergency department with a suspected acute coronary syndrome. AB - AIM: To describe the 10-year prognosis and risk indicators of death in women admitted to the emergency department with acute chest pain or other symptoms raising a suspicion of acute myocardial infarction (AMI). Particular interest was paid to women of 5-fold) increase in PTHrP mRNA half-life in TGFbeta1-treated samples when PTHrP mRNA lacked the 3'-untranslated region (3'-UTR). In contrast, the degradation of 3'-UTR-truncated PTHrP mRNA using the cell-free assay was not altered in vehicle-treated samples. UV cross linking of PTHrP mRNA and cytoplasmic proteins from cells treated with either vehicle or TGFbeta1 revealed numerous mRNA-binding proteins. TGFbeta1 treatment resulting in decreased binding of 33, 31, 27, 20 and 18 kDa binding proteins to the terminal coding region. These studies revealed that TGFbeta1-induced PTHrP mRNA stability might be, in part, the result of cis-acting sequences within the coding region of the PTHrP mRNA. PMID- 11911945 TI - Radicicol represses the transcriptional function of the estrogen receptor by suppressing the stabilization of the receptor by heat shock protein 90. AB - The estrogen receptor (ER) is a hormone-dependent transcription factor that belongs to the steroid/thyroid hormone receptor superfamily. Since the ER contributes to development and progression in human breast cancer, a number of studies have explored ways to inactivate this receptor. Previous studies have suggested that the 90-kDa heat shock protein (Hsp90) interacts with the ER, thus stabilizing the receptor in an inactive state. Here, we report that radicicol, an Hsp90-specific inhibitor, repressed estrogen-dependent transactivation of the ER as measured by pS2 gene transcription and a reporter gene encoding an estrogen responsive element. Furthermore, we showed that radicicol induced rapid degradation of ERalpha, while the amount of ubiquitinated ERalpha was increased. A proteasome inhibitor, LLnL, almost completely abrogated the radicicol-induced decrease in expression level, as well as in transcriptional activity of ERalpha. These results suggest that radicicol disrupts the ER-Hsp90 heterodimeric complex, thereby generating ERalpha that is susceptible to ubiquitin/proteasome-induced degradation. PMID- 11911946 TI - Interaction between arachidonic acid and cAMP signaling pathways enhances steroidogenesis and StAR gene expression in MA-10 Leydig tumor cells. AB - Previous studies have demonstrated that trophic hormone stimulation induced cyclic AMP (cAMP) formation and arachidonic acid (AA) release from phospholipids and that both these compounds were required for steroid biosynthesis and steroidogenic acute regulatory (StAR) gene expression in MA-10 mouse Leydig tumor cells. The present study further investigates the synergistic effects of the AA and cAMP interaction on steroidogenesis. To demonstrate cAMP-induced AA release, MA-10 cells were pre-loaded with 3H-AA and subsequently treated with dibutyryl cyclic AMP (dbcAMP). Stimulation with dbcAMP significantly induced AA release in MA-10 cells to a level 145.7% higher than that of controls. Lowering intracellular cAMP concentration by expressing a cAMP-phosphodiesterase significantly reduced human chorionic gonadotrophin (hCG)-induced AA release. The dbcAMP-induced AA release was inhibited significantly by the phospholipase A(2) (PLA(2)) inhibitor dexamethasone (Dex) and also by the protein kinase A (PKA) inhibitor H89, suggesting the involvement of PKA phosphorylation and/or PLA(2) activation in cAMP-induced AA release. The effect of the interaction between AA and cAMP on StAR gene expression and steroid production was also investigated. While 0.2 mM dbcAMP induced only very low levels of StAR protein, StAR mRNA, StAR promoter activity and steroid production, all of these parameters increased dramatically as AA concentration in the culture medium was increased from 0 to 200 microM. Importantly, AA was not able to induce a significant increase in steroidogenesis at any concentration when used in the absence of dbcAMP. However, when used in concert with submaximal concentrations of dbcAMP (0.05 mm to 0.5 mm), AA was capable of stimulating StAR gene expression and increasing steroid production significantly. The results from this study demonstrate that AA and cAMP act in a highly synergistic manner to increase the sensitivity of steroid production to trophic hormone stimulation and probably do so by increasing StAR gene expression. PMID- 11911947 TI - Leukemia inhibitory factor (LIF) promotes the primordial to primary follicle transition in rat ovaries. AB - In a sexually mature female, primordial follicles continuously leave the arrested pool and undergo the primordial to primary follicle transition. The oocytes increase in size and the surrounding squamous pre-granulosa cells become cuboidal and proliferate to form a layer of cuboidal cells around the growing oocyte. This development of the primordial follicle commits the follicle to undergo the process of folliculogenesis. When the available pool of primordial follicles is depleted reproductive function ceases and humans enter menopause. The current study examines whether leukemia inhibitory factor (LIF) promotes the primordial to primary follicle transition that initiates follicular development. Ovaries from 4 day-old rats were cultured in the absence or presence of LIF or neutralizing antibody to LIF. LIF treatment increased the proportion of follicles that initiated the primordial to primary follicle transition to 59%, compared to 45% in untreated cultured ovaries. The ability of LIF to induce primordial follicle development was enhanced to greater than 75% by the presence of insulin in the culture medium. Anti-LIF neutralizing antibody reduced the proportion of spontaneous developing primordial follicles. Immunocytochemical studies demonstrated higher levels of LIF protein in the granulosa and surrounding somatic cells of primordial and primary follicles compared to the oocyte. In contrast, later pre-antral and antral stage follicles showed LIF expression primarily in the oocyte. In granulosa and theca cell cultures LIF had no effect on cell proliferation. However, LIF treatment did increase expression of Kit ligand (KL) mRNA in cultured granulosa cells. KL has been shown to promote ovarian cell growth and induce primordial follicle development. LIF induction of KL expression may be involved in the actions of LIF to promote primordial to primary follicle transition. In summary, LIF treatment increased the primordial to primary follicle transition in cultured ovaries and LIF may interact with KL to promote primordial follicle development. PMID- 11911948 TI - Characterization of the human somatostatin receptor type 4 promoter. AB - Somatostatin (SRIF) exerts inhibitory effects on virtually all endocrine and exocrine secretions. Five distinct SRIF receptor subtypes (sst 1-5) have been identified. In contrast to the other subtypes, very little is known about specific functions of sst4. We investigated structure and regulation of the human sst4 gene. A genomic clone containing the 5' region of the sst4 gene was isolated. 1.5 kb of the promoter was sequenced and putative transcription factor binding sites were identified. The transcription start site was located 88 nucleotides upstream of the translation start site. A -984 sst4 promoter directed significant levels of luciferase expression in GH4 rat pituitary cells, Skut-1B endometrium cells, and BEAS-2B human bronchial epithelial cells, whereas only low activity was detected in JEG3 chorion carcinoma cells or COS-7 monkey kidney cells. A minimal -209 promoter allowed cell specific expression, its activity in COS-7 cells is not enhanced by co-transfection of the pituitary-specific transcription factor Pit-1. An enhancer element was localized between nt -459 and -984. We did not find any regulation of the sst4 promoter region analyzed by SRIF, forskolin, TPA, IGF-1, EGF, T3, glucocorticoids or 17beta-estradiol. These studies identify the 5' region of the sst4 gene. Furthermore, specific activity of the promoter in various cell lines is demonstrated. PMID- 11911949 TI - Intron-mediated expression of the human neuropeptide Y Y1 receptor. AB - The Neuropeptide Y (NPY) family of neuropeptides exert their function through a family of heptahelical G-protein coupled receptors regulating essential physiological processes. A 97 base pair intron (intron IV) intervenes the coding sequence of the human NPY Y1 receptor (hY1) gene and was found frequently retained at variable levels in poly A+ mRNA isolated from multiple human tissues. When included in hY1 expression vectors, either in its natural position or 5' of the hY1 cDNA, intron IV mediated a significant increase of both hY1 mRNA and corresponding functional receptor protein in transfected mammalian cells, implying an in vivo regulatory function of the endogenous intron. Our results further indicate that the nuclear history of the hY1 pre-mRNA influence ectopic hY1 production through post-transcriptional mechanisms and argues against intron IV acting as a transcriptional enhancer as well as the possibility that a putative hY1 related 5TM accessory protein encoded by the non-spliced hY1 mRNA would facilitate hY1 production on a post-translational level. PMID- 11911950 TI - Stimulation of the alpha-fetoprotein promoter by unliganded thyroid hormone receptor in association with protein deacetylation. AB - alpha-Fetoprotein (AFP) is a serum protein expressed during fetal life, the expression of which is shut off after birth. The activity of the mouse Afp gene promoter region comprised between -80 and -38 bp is regulated by the thyroid hormone receptor (T3R): negatively in the presence of T3 and positively in the absence of T3. The stimulating effect of unliganded T3R is, unexpectedly, antagonized by cofactors that have histone-acetyl-transferase activity, or by sodium butyrate, which inhibits histone acetylases (HDACs). The unliganded T3R stimulating activity effect is thus associated with protein deacetylation, contrary to the usual situation. In combination with previous results, our observations suggest that T3-mediated down regulation of the Afp promoter is due to T3-induced protein acetylation leading to loss of a nucleosomal structure (required for promoter activity) and chromatin opening. PMID- 11911951 TI - CDB-4124 and its putative monodemethylated metabolite, CDB-4453, are potent antiprogestins with reduced antiglucocorticoid activity: in vitro comparison to mifepristone and CDB-2914. AB - To obtain selective antiprogestins, we have examined the in vitro antiprogestational/antiglucocorticoid properties of two novel compounds, CDB-4124 and the putative monodemethylated metabolite, CDB-4453, in transcription and receptor binding assays and compared them to CDB-2914 and mifepristone. All four antiprogestins bound with high affinity to rabbit uterine progestin receptors (PR) and recombinant human PR-A and PR-B (rhPR-A, rhPR-B) and were potent inhibitors of R5020-induced transactivation of the PRE2-tk-luciferase (PRE2-tk LUC) reporter plasmid and endogenous alkaline phosphatase production in T47D-CO human breast cancer cells. None of these compounds exhibited agonist activity in these cells. Induction of luciferase activity was potentiated about five-fold by 8-Br-cAMP under basal conditions and to the same extent in the presence of the PR antagonists. Mifepristone bound to rabbit thymic glucocorticoid receptors (GR) with approximately twice the avidity of the CDB antiprogestins. Inhibition of GR mediated transcription of PRE2-tk-LUC was assessed in HepG2 human hepatoblastoma cells. Mifepristone exhibited greater antiglucocorticoid activity than CDB-2914, 4124, and 4453, about 12-, 22-, and 185-fold, respectively. Thus, while there was a good correlation between binding to PR and functional activity of these antiprogestins, GR binding was not predictive of their glucocorticoid antagonist activity. In agreement with our in vivo results, CDB-4124 and CDB-4453, as well as CDB-2914, are potent antiprogestins in vitro, but show considerably less antiglucocorticoid activity than mifepristone. PMID- 11911953 TI - Stanniocalcin from an ancient teleost: a monomeric form of the hormone and a possible extracorpuscular distribution. AB - Stanniocalcin (STC) is a homodimeric glycoprotein hormone implicated in calcium and phosphate regulation in both teleost fish and mammals. In the present study, immunostaining with salmon STC antiserum demonstrated that STC cells were localized in both the corpuscles of Stannius (CS) and in specific cells of the distal renal tubules of the silver arawana, Osteoglossum bicirrhosum, an ancient ray-finned fish (actinoptergian) and basal teleost (Order Osteoglossiforme). The morphology of these STC-immunoreactive kidney cells was similar to renal 'chloride' (mitochondrial-rich) cells. The immunoreactive renal cells were present in two of three other osteoglossiformes and absent in the gar, a nonteleost actinopterygian and the eel, another basal teleost. The arawana STC cDNA encodes a prehormone of 249 amino acids (aa) with a signal peptide of 31 aa and a mature protein of 218 aa. The deduced aa sequence of arawana STC shows 54 66% identity with other teleost STCs and 51-52% identity with mammalian STC-1. The deduced aa sequence of arawana STC contains ten cysteines, compared with 11 in teleost STC and in mammalian STC-1. The cysteine substitution occurs at the site of inter-monomeric disulfide linkage. Western blot analysis revealed a single 21 kDa band under non-reducing conditions, and a single band of 25 kDa under reducing conditions. These data indicate that arawana STC exists as a monomeric peptide. Northern blot analysis detected a 3.3 kb STC mRNA confined to the CS, with no hybridization signal in either the remaining kidney or in gut, muscle, brain and heart. The significance of the STC signal in cells of the renal tubules of arawana and two other Osteoglossiforme species requires further investigation. This is the first report of a monomeric form of STC in any vertebrate and the first evidence of STC in renal tubules of adult fish. PMID- 11911952 TI - Gender-dependent expression of alpha and beta estrogen receptors in human nontumor and tumor lung tissue. AB - Estrogen receptor (ER) expression in human lung has been understudied, particularly in light of its potential biological importance in the female lung cancer epidemic. Reverse transcription-polymerase chain reaction was used to probe mRNA expression of wild-type ERalpha and ERbeta and their splice variants in human bronchogenic tumor and adjacent nontumor specimens. In tumor tissue from 13 women and 13 men, ERalpha was expressed in 85% of women versus 15% in men [P=0.001]. ERbeta was expressed equally in tumors from women versus men [92% vs. 69%, P=ns]. Both ERalpha and beta forms were expressed simultaneously in the lung tumors of 77% of women versus 15% of men [P=0.005]. Among adjacent nontumor lung specimens, 31% of the women expressed ERalpha mRNA versus 0% of men [P=0.101], and 39% of women expressed ERbeta mRNA versus 31% of men [P=ns]; only one woman and no men expressed both ERalpha and beta in nontumor tissue. Females expressed ERalpha [P=0.017], ERbeta [P=0.013], and ERalpha+beta [P=0.002] more frequently in tumor versus nontumor tissue, whereas in males expression of ERalpha, beta and both alpha+beta was not clearly different for tumor versus nontumor tissue. In specimens expressing ERalpha mRNA, the transcript lacking exon 7 (delta7) was the major splice variant with varying contributions from the transcripts delta4, delta3+4, delta5 and others unidentified. Alternative splicing of ERbeta mRNA was observed, but not to as great an extent as for ERalpha mRNA. ERalpha promoter usage in tumors varied among individuals. When the ER receptors were co-expressed in tumors, ERalpha was quantitatively more abundant in the majority of cases than ERbeta. Within this small group of 26 patients, no correlation was found between age, smoking history, plasma nicotine, cotinine, estradiol concentrations or histopathologic type with tumor or nontumor estrogen receptor status of any type. However, several positive correlations imply that: (1) ERalpha expression occurs more often in the lungs of women than men; (2) ERbeta is expressed with approximately equal frequency in the lungs of both genders; and (3) tumors display a higher frequency of both receptor types than nontumors in women. We hypothesize that these putative gender-dependent differences in ERalpha and ERbeta expression could contribute unique phenotypic characteristics to lung cancer development or progression in women. PMID- 11911954 TI - Oxytocin inhibits LH-stimulated production of androstenedione by bovine theca cells. AB - Oxytocin secretion by bovine granulosa cells increases dramatically after the LH/FSH surge. We have shown that oxytocin stimulates progesterone secretion and inhibits FSH-stimulated estradiol secretion in vitro by granulosa cells from bovine preovulatory follicles obtained before the LH/FSH surge. To determine if oxytocin regulates LH-stimulated steroid production by bovine theca interna cells, theca cells were isolated from preovulatory follicles obtained before the LH surge and were cultured for 4 days in the presence or absence of LH (2 or 4 ng/ml), without or with graded doses of oxytocin (125-1000 ng/ml). LH increased accumulation of androstenedione and progesterone. Oxytocin inhibited LH stimulated androstenedione production, but had no effect on LH-stimulated progesterone production by cultured theca interna. The next objective was to determine if oxytocin regulates LH-stimulated steroidogenesis by modulating the levels of mRNA for steroidogenic enzymes and/or Steroidogenic Acute Regulatory protein (StAR). Low doses of LH alone increased the levels of mRNA for P450 17 alpha-hydroxylase (17 alpha-OH), 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and cytochrome P450 side-chain cleavage, but not for StAR. In contrast, the effects of oxytocin on LH-stimulated androstenedione production were not associated with changes in the levels of mRNA for steroidogenic enzymes or StAR. These results suggest that oxytocin may play a paracrine role in regulating the follicular/luteal phase shift in steroidogenesis by decreasing androstenedione secretion by theca cells of the ovulatory follicle and that this effect is not mediated by changes in the levels of mRNA for steroidogenic enzymes and StAR. PMID- 11911955 TI - Transcriptional repression of the rat steroidogenic acute regulatory (StAR) protein gene by the AP-1 family member c-Fos. AB - PGF2alpha, working via protein kinase C, may inhibit transcription of the StAR gene through negative regulatory factors. Administration of PGF2alpha increased c Fos mRNA with a corresponding reduction in StAR mRNA. A search of the rat StAR promoter revealed three putative AP-1 elements at bp positions -85, -187 and 1561, which demonstrated specific binding of c-Fos by mobility shift assays. Co transfection of c-Fos with the p-1862 StAR promoter caused a reduction in luciferase activity in the presence or absence of cAMP. Mutation of all three AP 1 sites in the p-1862 StAR promoter abolished c-Fos repression. Mutation of the proximal AP-1 site in the p-1862 StAR promoter reduced SF-1 mediated induction. This study is the first to demonstrate that c-Fos represses StAR gene transcription and adds to the current knowledge on the complex relationship that exists between SF-1 and c-Fos in the regulation of StAR activity. PMID- 11911956 TI - Molecular characterization of LH-beta and FSH-beta subunits and their regulation by estrogen in the goldfish pituitary. AB - The gonadal steroids, along with gonadotropin-releasing hormone (GnRH) are involved in the regulation of gonadotropin (GtH) production in vertebrates. Goldfish have an annual reproductive cycle, characterized by seasonal fluctuations in the circulating levels of the reproductive hormones, including 17beta-estradiol (E2). The purpose of the present study was to investigate the effect of E2 on basal and GnRH-induced GtH subunit (alpha, FSH-beta and LH-beta) gene expression in the goldfish pituitary. Northern analyses were performed to determine changes in steady state mRNA levels. Both in vivo and in vitro treatment with E2 resulted in a stimulation of all three GtH subunit mRNA levels, although a higher concentration was required for the stimulation of the FSH-beta subunit mRNA levels. The effect of E2 on GnRH-induced GtH mRNA level was also investigated and demonstrated that E2 influences the GnRH-induced GtH subunit mRNA levels in a seasonally dependent manner. Overall, the present results indicate that E2 stimulates GtH subunit mRNA levels directly at the level of the pituitary in a seasonally dependent manner in goldfish. PMID- 11911957 TI - Gonadotropin regulation of activin betaA and activin type IIA receptor expression in the ovarian follicle cells of the zebrafish, Danio rerio. AB - We have previously demonstrated that both activin and its receptors are expressed in the zebrafish ovary, suggesting paracrine roles for activin in the ovarian functions. Activin significantly stimulated zebrafish oocyte maturation in vitro, and this effect could be blocked by follistatin, an activin-binding protein. Interestingly, follistatin also blocked the stimulatory effect of gonadotropin (hCG) on the oocyte maturation. Taken together, these results have led to a hypothesis that the ovarian activin system may play a role in mediating the actions of gonadotropin in the ovary. To test this hypothesis, the present study was undertaken to investigate if gonadotropin has any effect on the expression of activin betaA subunit and activin type IIA (ActRIIA) receptor in the zebrafish ovary. A primary culture of zebrafish ovarian follicle cells was established in the present study, and the cultured cells expressed both activin betaA and ActRIIA receptor when assayed with RT-PCR. The primary culture consisted of three major types of cells, presumably the fibroblasts, the thecal cells and the granulosa cells, according to the morphological features, histochemical staining for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and RT-PCR for aromatase. Using a semi-quantitative RT-PCR with beta-actin as the internal control, we demonstrated that hCG significantly stimulated mRNA expression of both activin betaA and ActRIIA receptor in the cultured follicle cells in a time- and dose dependent manner. Treatment of the cells with hCG quickly increased the steady state mRNA levels of activin betaA and ActRIIA receptor, and the effect peaked at 2 h of treatment. The stimulatory effect of gonadotropin diminished with longer treatment and no effect was observed at 8 h of treatment. The effect of hCG also exhibited strong dose dependence when assayed at 2 h of treatment. The levels of activin betaA and ActRIIA receptor mRNA elevated with increasing dose of hCG; however, the effect significantly decreased at dosage higher than 15 IU/ml. Consistent with the stimulatory effect of gonadotropin on the expression of activin betaA and ActRIIA receptor, IBMX, forskolin and 8-Br-cAMP all significantly increased the mRNA levels of activin betaA and ActRIIA receptor. These results suggest that gonadotropin activates the activin system in the zebrafish ovary by increasing the expression of both activin and its receptors. PMID- 11911958 TI - Quantitation of the mRNA levels of Epo and EpoR in various tissues in the ovine fetus. AB - A partial cDNA of the sheep erythropoietin receptor (EpoR) was obtained and used in real-time PCR to quantitate mRNA levels in placenta, liver and kidney throughout development (term=150 days). This was compared with Epo mRNA levels in the same tissues. Both Epo and EpoR mRNA were present in the placenta throughout gestation at low levels from 66 days onwards and these did not vary throughout gestation. Compared with the expression levels in the placenta, the levels of EpoR gene expression in the liver at 66, 99 and 140 days were, median (range)-288 (120-343), 278 (63-541) and 7 (3-15), respectively, reflecting the disappearance of erythropoiesis after 130 days. Low levels of EpoR gene expression were seen in the kidney at 3 (2-5), 5 (2-7), and 7 (2-10) times that in the placenta at 66, 99, and 140 days, respectively. By hybridization histochemistry the EpoR mRNA was located in the proximal tubular cells of the mesonephros and metanephros at 42 days. Epo mRNA levels in the kidney were 215 (116-867), 528 (113-765) and 46 (15 204) times those in the placenta at 69, 99, and 140 days, respectively. In the liver at the same ages the concentrations of mRNA were lower than in the kidney, the liver/placenta ratios being 50 (11-90), 17 (3-39), 9 (5-14). At 130 days Epo/EpoR levels in the hippocampus were 6+/-3 and 8+/-3 times that in the term placenta, respectively. These studies demonstrate that the ovine placenta expresses the Epo gene from at least 66 days of gestation. However, gene expression levels are very low compared with those in the liver and kidney, and even the hippocampus. PMID- 11911959 TI - Identification of leptin receptors in human breast cancer: functional activity in the T47-D breast cancer cell line. AB - To evaluate whether leptin plays a putative role in breast tumorigenesis, we studied the expression of its long and short receptor isoforms in various tumoral breast tissues. We applied semiquantitative RT-PCR method to RNA extracted from 20 breast cancer biopsies and two human breast cancer cell lines (T47-D and MCF 7). Our results showed the expression of both leptin receptor transcripts in all tumoral tissues examined. By in situ hybridization experiments, we localized leptin receptors in proliferating epithelial cells. Study of leptin effects on human breast cancer cells growth was performed by [3H]-thymidine incorporation method and colorimetric MTT assay. We demonstrated that leptin (50-100 ng/ml) significantly stimulates proliferation of the human breast cancer cell line T47-D (P<0.05). Western blot analysis indicated that leptin induces a time-dependent activation of mitogen-activated protein kinases (MAPKinase) 1 and 2 in T47-D cell line. Moreover, the specific MAPK-inhibitor PD 98059 blocked cell proliferation induced by leptin. In conclusion, we demonstrate that leptin receptors are expressed in breast cancer and that leptin induces proliferation in the T47-D cell line via activation of the MAPKinases pathway. These data suggest that leptin and its receptors may be implicated in mammary cell proliferation and in breast cancer pathogenesis. PMID- 11911960 TI - 1,25-Dihydroxyvitamin D3 selectively translocates PKCalpha to nuclei in ROS 17/2.8 cells. AB - We have investigated protein kinase C (PKC) regulation by 1,25-(OH)2D3 in the rat osteosarcoma cell line ROS 17/2.8 since previous reports have implicated PKC in the 1,25-(OH)2D3-mediated regulation of osteocalcin gene expression (J. Biol. Chem. 267 (1992) 12562; Endocrinology 136 (1995) 5685). Here we report that 1,25 (OH)2D3 increased PKCalpha, but not PKCbetaI, epsilon or zeta, levels in the nuclear fraction in a time-dependent manner. Unlike PMA, 1,25-(OH)2D3 did not alter the association of any of the expressed PKC isoenzymes with the plasma membrane. Treatment with 20 nM 1,25-(OH)2D3 for 15 min, 30 min, 1 h and 24 h increased PKCalpha levels in the nuclear fraction by 2.3- to 2.6-fold. Nuclear PKCalpha expression was also increased with doses of 1,25-(OH)2D3 as low as a 0.05 nM. 1,25-(OH)2D3-mediated stabilization of osteocalcin mRNA (Arch. Biochem. Biophys. 332 (1996) 142) was inhibited with bisindolylmaleimide treatment, suggesting that PKCalpha may be involved in the 1,25-(OH)2D3-mediated regulation of osteocalcin gene expression. PMID- 11911961 TI - The influence of alpha1-acid glycoprotein (orosomucoid) and its glycoforms on the function of human thyrocytes and CHO cells transfected with the human TSH receptor. AB - Local immunological reactions might influence the structure of alpha1-acid glycoprotein (AGP, orosomucoid) leading to a pathological condition in e.g. the thyroid. The aim of this study was to investigate whether the AGP molecule had a direct effect on thyroid cell function in vitro. The influence of AGP and its three glycoforms, TSH (1.0 U/l), serum samples and several sugars (methyl mannose, methyl-glycoside, N-acetyl-D-galactose, N-acetyl-D-glycoside, neuramidase) were studied with respect to their influence on the function of the Chinese Hamster Ovary (CHO) cell line transfected with the human TSH receptor (hTSHr) and on human thyroid follicular epithelial cells (TFEC) in secondary cultures. We found that low concentrations of AGP (0.001-0.05 microg/l) stimulated while high concentrations of AGP (0.25-1.0 microg/l) inhibited cAMP accumulation in both cell systems (n=24, P<0.0002). In CHO cells (JP26) and TFEC glycoforms 1 (n=9), 2 (n=12) or 3 (n=11) significantly inhibited the TSH stimulated cAMP production, respectively, compared to controls (P<0.0001) and was partially reversed by mannose (P<0.0004). Control CHO cells (JP02) without the hTSHr showed no response. The specificity of the reaction was further confirmed by binding of biotinylated glycoforms and streptavidin conjugated FITC to both cell systems. This is the first report demonstrating that AGP and/or its glycoforms affects thyroid cell function in vitro and that it does so by influencing the second messenger cAMP probably by interacting directly with the TSH receptor. PMID- 11911962 TI - Antagonism of activin by inhibin and inhibin receptors: a functional role for betaglycan. AB - Activin and inhibin research has provided important insight into reproductive physiology as well as many areas involving regulation of cell growth, differentiation and function. Progress in understanding the roles of these hormones in various cell and tissue types has been complimented by novel discoveries at the molecular level that have shed light on ligand/receptor interactions, signaling mechanisms and regulation. While the receptors and signaling pathway for activin are now well characterized, the molecular basis for inhibin action has remained relatively unclear. Here we summarize recent advances in understanding inhibin's mode of action focusing on our recent identification of betaglycan as an inhibin co-receptor capable of mediating inhibin action. PMID- 11911963 TI - In vitro induction of the anticarcinogenic marker enzyme, quinone reductase, in human hepatoma cells by food extracts. AB - The effect of vegetable extracts on the activity of the anticarcinogenic phase II marker enzyme, quinone reductase (QR), was investigated by using human Hep G2 cells as the model system. Hep G2 cells were less sensitive than murine Hepa1c1c7 cells to QR-inducible compounds such as tert-butylhydroquinone which have been widely used to examine the QR-inducing activity of the compounds. However, among 45 different vegetable samples, an extract of ashitaba clearly induced QR activity in Hep G2 cells. Ashitaba is therefore considered to have contained certain substances that could induce QR activity, and such induction may play a role in the anticarcinogenic action of vegetables. PMID- 11911964 TI - Roscovitine inhibits ongoing DNA synthesis in human cervical cancer. AB - The effect of roscovitine, a purine analogue and cyclin dependent kinase inhibitor, on DNA synthesis rate in tissue mini-units obtained from human cervical cancers was investigated. Roscovitine (100 microM) gave a DNA synthesis rate inhibition by 61% (P<0.0001; range 23-93%) within 30 min of incubation. This inhibitory effect was concentration-dependent. The results suggest that the inhibition of tumor DNA synthesis rate is due to a direct effect on the DNA synthesis machinery via presently unknown mechanisms. In addition, the potential application of CDKs inhibitors as preventive agents is discussed. PMID- 11911966 TI - Antitumoral effects of recombinant adenovirus YKL-1001, conditionally replicating in alpha-fetoprotein-producing human liver cancer cells. AB - Selectively replicating recombinant adenovirus has emerged as a novel strategy for the treatment of incurable human cancers. One of the major characteristics of hepatocellular carcinoma is the transcriptional reactivation of alpha-fetoprotein (AFP). In this study, we evaluated the liver cancer-specific oncolytic potential of E1B 55kDa-deleted recombinant adenovirus (YKL-1001), which retained other E1 genes driven by the AFP promoter. Transient transfection study using luciferase indicated the selective activation of the AFP promoter only in human liver cancer cells secreting AFP (HepG2, Hep3B). YKL-1001 induced both cytopathic effects exclusively in AFP-positive liver cancer cells and the growth inhibition of pre established Hep3B xenografts. Finally, hematoxylin-eosin staining and the immunohistochemistry to the adenoviral hexon showed a large distributed necrotic area and this implied a wide spread of YKL-1001. Therefore, the present study demonstrated that YKL-1001 holds significant promise as an oncolytic agent for hepatocellular carcinoma. PMID- 11911965 TI - Overexpression of cyclooxygenase-2 (COX-2) in human primitive neuroectodermal tumors: effect of celecoxib and rofecoxib. AB - In this study the role of cyclooxygenase-2 (COX-2) in primitive neuroectodermal tumor (PNET) the most malignant brain tumors of childhood was investigated. COX-2 expression in human brain tumor biopsy samples (seven/seven) was about 6-8-fold higher than normal brain tissue and several PNET cell lines also express COX-2. The effect of selective COX-2 inhibitors, celecoxib and rofecoxib on the growth of two PNET cell lines (DAOY and PFSK) was determined. Celecoxib was more potent than rofecoxib in suppressing cell growth. Growth inhibition by celecoxib and rofecoxib was independent of Bcl-2 expression. Celecoxib suppressed the expression of Akt and activated the caspase-3 in DAOY and PFSK, whereas rofecoxib did not have such an effect. PMID- 11911967 TI - O6-Alkylguanine-DNA-alkyltransferase activity in peripheral leukocytes, smoking and risk of lung cancer. AB - The level of activity of O6-alkylguanine-DNA-alkyltransferase (AGT), a DNA repair enzyme, in blood lymphocytes may be a marker of susceptibility to lung cancer. We measured the AGT activity level, expressed as pmoles of repaired bases/mg protein, in leukocytes of 153 lung cancer cases (of whom 80 were never smokers) and 106 controls (76 never smokers) enrolled in eight centres from seven countries. Subjects were interviewed with respect to active smoking and exposure to environmental tobacco smoke (ETS). Among never smokers, the odds ratios (ORs) of lung cancer were 1.3 (95% confidence interval 0.5-3.9), 1.5 (0.6-4.1) and 1.4 (0.5-3.8) in quartiles of decreasing AGT activity level, as compared to the upper quartile (P value of test for linear trend 0.6). Corresponding ORs among smokers were 3.4 (0.9-13), 2.0 (0.5-8.3) and 0.4 (0.1-1.6) (P value of test for linear trend 0.4). No interaction was suggested between AGT activity level and either cumulative smoking or exposure to ETS. Reduced AGT activity was not clearly associated with increased lung cancer risk in either smokers or non-smokers. However, the small size of our study argues for a prudent interpretation of our results. PMID- 11911968 TI - Influence of cytochrome P450 CYP2C9 genotypes in lung cancer risk. AB - Cytochrome P-450 CYP2C9 enzyme is involved in the metabolism of xenobiotics and carcinogens. Variant alleles CYP2C9*2 and CYP2C9*3 encode enzymes with altered properties, and CYP2C9*2 has been related to lung cancer risk. The frequency of CYP2C9 variant alleles was analyzed in genomic DNA from 104 patients with lung cancer and in 197 healthy controls. No statistically significant differences in CYP2C9 genotypes, allele frequencies or predicted phenotypes were observed between overall patients and controls. Unconditional logistic regression for the interaction smoking/genotype indicate P values of 0.66 and 0.62 for carriers of CYP2C9*2 and CYP2C9*3, respectively, versus individuals without these mutations. The P values for the same interaction in carriers of one and two mutated genes were 0.56 and 0.67, respectively, versus individuals homozygous for non-mutated genes. Independent analyses of histological types of lung cancer also indicated the absence of statistically significant differences as compared to healthy subjects. Association between CYP2C9 polymorphism and lung cancer risk was not identified in this study. PMID- 11911969 TI - CYP17 genetic polymorphism in endometrial cancer: are only steroids involved? AB - Initiation and/or promotion of endometrial cancer is known to be associated with estrogen and androgen (androstenedione) excess as well as with hyperinsulinemia/insulin resistance. It is possible that some allelic polymorphisms of the genes involved in steroidogenesis or steroid metabolism contribute to endometrial cancer susceptibility. We evaluated here the role of CYP17 biallelic (MspAI) polymorphism in 114 endometrial cancer patients compared with 182 healthy women. Our data demonstrated that A2/A2 CYP17 genotype, considered on the basis of initial breast cancer studies as 'unfavorable', was under-represented in endometrial cancer group (odds ratio 0.48, 95% confidence interval 0.25-0.89) that confirmed results of two other recent investigations. Carriers of this genotype were characterized by having lower blood insulin (by 120 min of oral glucose tolerance test 36.7+/-3.9 microU/ml vs. 90.4+/-16.7 microU/ml in postmenopausal women with A1/A1 genotype, P=0.04) and C-peptide levels (after night fasting 575.2+/-78.3 pg/ml vs. 978.9+/-115.7 pg/ml, respectively, P=0.04). No significant difference was found between the mean concentrations of testosterone, dehydroepiandrosterone sulfate and estradiol concentrations in patients-carriers of separate CYP17 genotypes. Thus, CYP17 polymorphism (namely, carrying the 'normal' A1/A1 genotype) might be one of the risk factors for endometrial cancer development. A1/A1 CYP17 variant may be associated with untraditional (non-steroidal) pathways that calls for corresponding preventive measures in high-risk groups. PMID- 11911970 TI - Molecular and immunohistochemical analysis of signaling adaptor protein Crk in human cancers. AB - Crk is a signaling adaptor protein which is mostly composed of SH2 and SH3 domains, and has been shown to play a pivotal role in cell proliferation, differentiation, and migration. Because Crk was originally isolated as an avian sarcoma virus CT10 encoding oncoprotein v-Crk, we examined a potential role for c Crk in the carcinogenesis of human cancers. First, to analyze gene mutations of c Crk, we isolated a human bacterial artificial chromosome clone containing Crk genome and exon/intron structures. However, polymerase chain reaction-single strand conformation polymorphism methods failed to show any genomic mutations in the Crk exon which could be related to carcinogenesis. Second, immunohistochemical analysis of c-Crk-II demonstrated that the levels of c-Crk-II were significantly elevated in most of the tumors, particularly in the colon and lung cancers. Furthermore, immunoblot analysis using human lung cancer cell lines revealed that the expression levels of c-Crk-II were correlated to growth rates of cells. The elevated expression levels of c-Crk-II might be related to the development of human cancers. PMID- 11911972 TI - Induction of protein catabolism and the ubiquitin-proteasome pathway by mild oxidative stress. AB - Muscle wasting in cancer cachexia is associated with increased levels of malondialdehyde (MDA) in gastrocnemius muscles, suggesting an increased oxidative stress. To determine whether oxidative stress contributes to muscle protein catabolism, an in vitro model system, consisting of C2C12 myotubes, was treated with either 0.2 mM FeSO4, 0.1 mM H2O2, or both, to replicate the rise in MDA content in cachexia. All treatments caused an increased protein catabolism and a decreased myosin expression. There was an increase in the proteasome chymotrypsin like enzyme activity, while immunoblotting showed an increased expression of the 20S proteasome alpha-subunits, p42, and the ubiquitin-conjugating enzyme, E214k. These results show that mild oxidative stress increases protein degradation in skeletal muscle by causing an increased expression of the major components of the ubiquitin-proteasome pathway. PMID- 11911971 TI - Expression of apoptotic regulators and their significance in cervical cancer. AB - Insufficient apoptosis is implicated in many human cancers, including cervical carcinoma. The objectives of this study were to explore changes of apoptosis regulating gene expression and their clinical significance in cervical cancer. The expression of apoptosis-regulating genes, including five Bcl-2 family and two caspase family members, was evaluated in 43 cervical invasive squamous cell carcinomas, using immunohistochemistry. Specimens in which >or=10% of the neoplastic cells showed cytosolic immunoreactivity were considered to be immunopositive. Results were correlated with clinico-pathologic characteristics of the subjects. All seven apoptotic regulators examined were positive in a proportion of the tumors. The percentage of cases expressing Bax was higher in the patients without evidence of disease after treatment than in the patients alive with disease or who died of disease (P<0.05). A significant difference in disease-free survival was detected between Bax-positive and -negative groups (P<0.05), and in overall survival between Mcl-1-positive and -negative groups (P<0.05). Significant association between the seven markers tested was only found for caspase 3 and Bak immunoreactivity in cervical carcinoma (P<0.05). The results demonstrate expression of multiple apoptosis-modulating proteins in cervical cancer. There appears to be complex regulation of apoptosis protein levels in association with clinical behavior of cervical squamous cell carcinoma. PMID- 11911973 TI - Association of allelic loss at 8p22 with poor prognosis among breast cancer cases treated with high-dose adjuvant chemotherapy. AB - To identify specific allelic losses that might correlate with postoperative mortality of breast cancer patients treated with high-dose adjuvant chemotherapy consisting of cyclophosphamide, methotrexate and fluorouracil, we examined tumors from a cohort of 150 such patients, who were followed clinically for 5 years postoperatively, for allelic losses (loss of heterozygosity, LOH) among 18 microsatellite markers throughout the genome. Patients whose tumors had lost an allele at 8p22 had significantly higher risks of mortality than those whose tumors retained both alleles at those loci. At 8p22, the 5-year mortality rate was 31% among patients with losses vs. 8% with retention (P=0.0354). No other region showed correlation between LOH and prognosis. The data indicate that LOH at 8p22 is a significant predictor of postoperative mortality for breast cancer patients who received high-dose postoperative adjuvant chemotherapy. Thus, LOH at 8p22 can serve as a negative prognostic indicator to guide postoperative management of patients. PMID- 11911974 TI - Distinct lymphatic spread of endometrial carcinoma in comparison with cervical and ovarian carcinomas. AB - The distribution of metastatic pelvic lymph nodes (PLNs) and aortic lymph nodes (ALNs) in 27 node-positive endometrial carcinomas (ECs) was analyzed in comparison with that in 25 node-positive cervical carcinomas (CCs) and 58 node positive ovarian carcinomas (OCs). All patients underwent systematic pelvic and aortic lymphadenectomy. Lymph nodes were classified into the five subgroups: ALN above the inferior mesenteric artery (IMA; A1), ALN below the IMA (A2), the common iliac and sacral LNs (P1), the internal and external iliac LNs and obturator LNs (P2) and the suprainguinal LNs (P3). EC was similar to CC in that metastases to P2 were more frequent compared to A1 or A2, whereas EC and OC shared a common feature in that A1, A2 and P2 were involved at high rates. ALN metastases were significantly associated with P1 positivity in both EC and CC (P<0.05), but not in OC. However, the incidence of both ALN and PLN metastases in EC (67%) was similar to that in OC (61%), being much higher than that in CC (36%). ALN involvement alone was observed in 7% for EC, 0% for CC and 21% for OC. Based on the distribution of LN metastases, it appears that CC metastasizes primarily to PLN, whereas OC metastasizes almost equally to both PLN and ALN. Interestingly, EC can directly metastasize to both PLN and ALN with PLN metastases being dominant, a distinct lymphatic spread pattern better viewed as being somewhere between CC and OC. PMID- 11911976 TI - The interaction between leading and trailing limbs during stopping in humans. AB - While the initiation of gait has been well studied, the mechanisms of stopping the forward progression of the center of mass have received less attention. The purpose of this current experiment was to examine the effects of constraining cadence on how the body is brought to a stop. Lower limb electromyogram (gluteus medius, hamstrings and soleus (SOL)) and force plate data were compared between two gait conditions, walking and planned stopping, at three different cadences (100, 125 and 150% of normal cadence). As cadence increased, the onset of muscle activity prior to heel-strike decreased for all muscles except SOL. SOL activity was evident prior to heel-strike during stopping and after heel-strike during walking. As cadence increased, onset times before heel-strike were shorter in stopping and SOL became active more rapidly after heel-strike in walking. The normalized duration of muscle activity remained invariant as cadence increased. This activity was always longer under the lead limb during stopping at each cadence. This was reflected by increases in the braking forces produced by the leading limb. The rate at which force was generated was not different between gait conditions but increased with increasing cadence. Thus, subjects relied less on the trailing limb and more on the leading limb as cadence increased. PMID- 11911975 TI - Resistance to diverse apoptotic triggers in multidrug resistant HL60 cells and its possible relationship to the expression of P-glycoprotein, Fas and of the novel anti-apoptosis factors IAP (inhibitory of apoptosis proteins). AB - We studied the human HL60 leukemia cell line and its multidrug resistant (MDR) variant HL60R. In contrast to the HL60, HL60R showed an inability to undergo apoptosis from doxorubicin (Dox) or other different stimuli, including cisplatin, Fas ligation and serum withdrawal. HL60R cells lost surface Fas expression, but we found no evidence that Fas/FasL mediates the apoptotic effects of Dox in HL60. P-glycoprotein (P-gp) did not seem to play a major role as a specific inhibitor of apoptosis. In fact, the P-gp inhibitor verapamil reversed only partially the resistance to Dox-induced apoptosis of the MDR cells. In addition, it did not modify the rate of apoptosis induced from the other stimuli in the same cells. The expression of p53 or Bcl-2 was not different between HL60 and HL60R. However, in HL60R there was an increase in the mRNAs of inhibitory of apoptosis proteins (IAPs) like neuronal apoptosis inhibitory protein (NAIP), c-IAP-2 and survivin. Treatment with Dox or serum starvation strongly down-regulated X-linked IAP and survivin mRNAs in HL60. Cisplatin decreased NAIP and survivin mRNAs in the same cells. However, in HL60R the levels of these IAP mRNAs were much less affected by the treatments. These results support that IAPs may be involved in tumor resistance to chemotherapeutic drugs or other apoptotic agents. PMID- 11911977 TI - Post-tetanic depression of GABAergic synaptic transmission in rat hippocampal cell cultures. AB - The effect of tetanic stimulation (30 Hz, 4 s) on evoked GABAergic inhibitory postsynaptic currents (IPSCs) was studied in cell cultures of dissociated hippocampal neurons with established synaptic connections. It was found that tetanic stimulation elicited post-tetanic depression (PTD) of the evoked IPSCs with a duration of more than 50 s in about 60% of the connections tested; post tetanic potentiation was induced in 25% of the connections. We propose that the opposite effects of tetanization on IPSC amplitude are due to differences in the type of the interneuron that was tetanized. Since PTD in our experiments was usually accompanied by changes in the IPSC coefficient of variation and changes of a paired pulse depression, which are thought to reflect presynaptic mechanisms of modulation, we suggest that part of the PTD is due to a presynaptic mechanism(s). PMID- 11911978 TI - Novel effect of vitamin K(1) (phylloquinone) and vitamin K(2) (menaquinone) on promoting nerve growth factor-mediated neurite outgrowth from PC12D cells. AB - The nerve growth factor (NGF)-potentiating effect of K vitamins on PC12D cells was investigated. Treatment of PC12D cells with vitamin K(1) or K(2) in the presence of NGF significantly enhanced the proportion of neurite-bearing cells and acetylcholinesterase activity compared with NGF treatment alone. The K vitamins-enhanced neurite outgrowth on PC12D cells was completely blocked by a protein kinase A (PKA) inhibitor or mitogen-activated protein kinase (MAPK) kinase inhibitor PD98059, whereas a protein kinase C inhibitor chelerythrine chloride did not significantly inhibit the enhancing effect of the K vitamins. These results suggest that the K vitamins enhance neurite outgrowth via the activation of PKA and MAPK-mediated signaling pathways in PC12D cells. PMID- 11911979 TI - Event-related theta power increases in the human EEG during online sentence processing. AB - By analyzing event-related changes in induced band power in narrow frequency bands of the human electroencephalograph, the present paper explores a possible functional role of the alpha and theta rhythms during the processing of words and of sentences. The results show a phasic power increase in the theta frequency range, together with a phasic power decrease in the alpha frequency range, following the presentation of words in a sentence. These effects may be related to word processing, either lexical or in relation to sentence context. Most importantly, there is a slow and highly frequency-specific increase in theta power as a sentence unfolds, possibly related to the formation of an episodic memory trace, or to incremental verbal working memory load. PMID- 11911980 TI - Non-rapid-eye-movement sleep propensity after sleep deprivation in human subjects. AB - The circadian modulation of occurrence of non-rapid-eye-movement sleep (NREM) was investigated in 37 volunteers under dim-light conditions after 24-h total sleep deprivation using a 26-h 10/20-min ultra-short sleep-wake schedule. The propensity of NREM showed rapid increase followed by gradual decrease during the subjective day and nocturnal bouts during the subjective night coinciding with melatonin production. The mean propensity of NREM during the subjective day was smaller than that during the subjective night, even though the sleep debt due to the 24-h sleep deprivation would have enhanced NREM more strongly during the subjective day than that during the subjective night. These results suggest that the occurrence of NREM sleep is modulated by a circadian pacemaker. PMID- 11911981 TI - Effects of fluctuations on electrical properties of gap-junction connected cells in the turtle retina. AB - Electrical properties of gap-junction connected cells (input voltage and length constant) are shown to depend strongly on fluctuations in membrane and contact conductances. This opens new possibilities and incorporates a further difficulty to the analysis of electrophysiological data, since four, instead of two, parameters (the average values and the magnitude of fluctuations of the two conductances) have to be used in fitting the experimental data. The discussion is illustrated by investigating the effects of dopamine on signal spreading in horizontal cells of turtle retina, assuming a linear cell arrangement. It is shown that while a standard fitting with the average values of the two conductances leads to the conclusion that both are equally affected by dopamine, including fluctuations allows fitting the data by varying just the average contact conductance plus the magnitude of fluctuations. PMID- 11911982 TI - Chronic cervical spinal sensory denervation reveals ineffective spinal pathways to phrenic motoneurons in the rat. AB - We hypothesized that pretreatment with chronic cervical dorsal rhizotomy (CDR; C(3)-C(6)) would reveal ineffective crossed spinal pathways to phrenic motoneurons. Anesthetized CDR (1 week post-rhizotomy) and control rats were spinally hemisected at C(2), and phrenic potentials were evoked by stimulating the ventrolateral funiculus contralateral and rostral to hemisection. Phrenic potentials contralateral to the stimulating electrode were evoked at lower stimulus currents (CDR=640 +/- 46 microA; control=900 +/- 50 microA; P<0.05) and potential amplitude was significantly greater in CDR versus control rats (P<0.05). The serotonin receptor antagonist methysergide (4 mg/kg, i.v.) had no effect on the crossed phrenic potential amplitude (91+/-17% of control at 800 microA; P>0.05). Thus, CDR enhances crossed phrenic pathways but serotonin receptor activation is not necessary to maintain this effect. PMID- 11911983 TI - Thalamic gray matter changes in unilateral Parkinsonian resting tremor: a voxel based morphometric analysis of 3-dimensional magnetic resonance imaging. AB - The thalamus is assumed to be involved in the generation of Parkinsonian tremor. Ten patients with tremor-dominant idiopathic Parkinson's disease (IPD) and strictly unilateral resting tremor were investigated by cerebral high-resolution 3-dimensional magnetic resonance imaging (MRI). MRI data were analyzed by an observer-independent morphometric technique, voxel-based morphometry (VBM). For VBM, MRI data were automatically normalized and segmented, then gray matter volumes were analyzed on a voxel-by-voxel basis in comparison to an age-matched control group using Statistical Parametric Mapping (SPM99). Highly significant structural changes, i.e. locally increased gray matter concentrations (P<0.001), were found in the nucleus ventralis intermedius (VIM) of the thalamus contralateral to the tremor side and were significantly covariant with tremor amplitudes. On the one hand, these changes were localized in close vicinity to a thalamic focal hypermetabolism as revealed by a previous positron emission tomography study in unilateral Parkinsonian tremor patients. On the other hand, the localization of the focal structural changes in VIM corresponds with the generally accepted target area of tremor surgery in IPD. PMID- 11911984 TI - Effects on splicing and protein function of three mutations in codon N296 of tau in vitro. AB - Three Mutations were recently reported in the same codon (N296) in exon 10 of the tau gene. Two of these mutations, N296N and N296H, lead to a clinical syndrome similar to autosomal dominant fronto-temporal dementia with Parkinsonism linked to chromosome 17. In contrast the third mutation, delN296, gives rise to atypical progressive supranuclear palsy in individuals homozygous for the mutation, but in heterozygous individuals this mutation is incompletely penetrant and associated with a phenotype similar to idiopathic Parkinson's disease. Functional assays were employed to determine the effects of these mutations on alternative splicing of exon 10, on microtubule assembly and self-aggregation of recombinant tau protein. We demonstrate that these mutations exhibit a spectrum of potentially pathogenic changes in tau function, and provide insight into the possible cause of the incompletely penetrant phenotype of the delN296 mutation. PMID- 11911985 TI - Ultrastructure of alpha-synuclein-positive aggregations in U373 astrocytoma and rat primary glial cells. AB - Abnormal alpha-synuclein-positive glial cytoplasmic inclusions are found in Parkinson's disease, multiple system atrophy and dementia with Lewy bodies. We have recently developed an in vitro model of alpha-synuclein-immunoreactive aggregations in U373 astrocytoma cells. We have additionally overexpressed wild type and a C-terminally truncated form of alpha-synuclein in primary rat glial cells. Astrocytes and oligodendrocytes were found to form alpha-synuclein positive aggregations in vitro perinuclearly or in the processes of the cells. The morphological studies presented here demonstrate that the aggregations we have observed in vitro are not limited by a membrane but have unclear borders. They have an amorphous dense core that is intensely alpha-synuclein immunopositive and a predominantly filamentous halo around. Mainly filamentous structures at the border area between the halo and the core are alpha-synuclein immunoreactive. We conclude that this in vitro model of alpha-synuclein-positive glial aggregations mimics the morphology of the abnormal glial inclusions described in neurodegenerative disorders and could be a suitable model for studying their role in the pathogenesis of these diseases. PMID- 11911987 TI - Lipofuscin and Abeta42 exhibit distinct distribution patterns in normal and Alzheimer's disease brains. AB - Our recent study has provided evidence that Abeta42, a 42 amino acid fragment of the amyloid precursor protein, accumulates intracellularly in vulnerable neurons. This study appears to show that neurons lyse and form dense-core amyloid plaques in Alzheimer's disease (AD) entorhinal cortex. Previous studies have suggested that intracellular Abeta42 co-localizes with lipofuscin in neurons and those increased levels of lipofuscin and Abeta42 are associated with AD. Other studies have questioned this relationship and suggested that beta-amyloid and lipofuscin are not co-localized and that their levels are independent of one another in AD and age-matched control tissues. In an effort to resolve this controversy, we investigated the relative spatial relationship of intracellular Abeta42 and lipofuscin in AD brains tissue using a novel combined immunohistochemical:histochemical staining protocol. Our results show separate and distinct localization patterns of Abeta42 and lipofuscin in neurons and amyloid plaques. PMID- 11911986 TI - Human otolith-ocular reflexes during off-vertical axis rotation: effect of frequency on tilt-translation ambiguity and motion sickness. AB - The purpose of this study was to examine how the modulation of tilt and translation otolith-ocular responses during constant velocity off-vertical axis rotation varies as a function of stimulus frequency. Eighteen human subjects were rotated in darkness about their longitudinal axis 30 degrees off-vertical at stimulus frequencies between 0.05 and 0.8 Hz. The modulation of torsion decreased while the modulation of horizontal slow phase velocity (SPV) increased with increasing frequency. It is inferred that the ambiguity of otolith afferent information is greatest in the frequency region where tilt (torsion) and translational (horizontal SPV) otolith-ocular responses crossover. It is postulated that the previously demonstrated peak in motion sickness susceptibility during linear accelerations around 0.3 Hz is the result of frequency segregation of ambiguous otolith information being inadequate to distinguish between tilt and translation. PMID- 11911988 TI - The striatal dopaminergic deficit is dependent on the number of mutant alleles in a family with mutations in the parkin gene: evidence for enzymatic parkin function in humans. AB - Autosomal recessive parkinsonism associated with mutations in the parkin gene represents a monogenic form of hereditary parkinsonism. We performed [(18)F]6 fluorodopa (FDOPA) positron emission tomography as a measurement of the nigrostriatal dopaminergic system as well as extensive haplotype analysis of the PARK 2 gene locus in 14 subjects with parkin mutations. In parkin subjects, the reduction of striatal FDOPA uptake increased with the number of mutated alleles and was also slightly obvious in asymptomatic parkin gene carriers in the heterozygous state. The abnormal FDOPA uptake pattern in parkin patients did not significantly differ from that of sporadic Parkinson's disease. Our data are in agreement with an enzymatic dysfunction of the gene's translational product, which has been shown to promote protein degradation as an ubiquitin-protein ligase. Thus, parkinsonism in parkin gene carriers may be related to abnormal nigral protein accumulation in the presence of a suprathreshold enzyme dysfunction. PMID- 11911989 TI - Melatonin improves deferoxamine antioxidant activity in protecting against lipid peroxidation caused by hydrogen peroxide in rat brain homogenates. AB - Deferoxamine (DF) is an antioxidant molecule because of its ability to chelate iron. This study compared the ability of DF alone or in combination with melatonin, 5-methoxytryptophol or pinoline in preventing lipid peroxidation due to hydrogen peroxide (H(2)O(2)) in rat brain homogenates. Malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA) in the homogenates were measured as indices of lipid peroxidation. Incubation of homogenates with DF reduced, in a dose dependent manner, MDA+4-HDA formation due to H(2)O(2). When melatonin, 5 methoxytryptophol or pinoline were added to the incubation medium, the efficacy of DF in preventing lipid peroxidation was enhanced. These cooperative effects between DF, melatonin, and related pineal products may be important in protecting tissues from the oxidative stress due to iron overload. PMID- 11911990 TI - Involvement of rat lateral septum-acetylcholine pressor system in central amygdaloid nucleus-emotional pressor circuit. AB - There is an emotional pressor circuit composed of nuclei controlling emotion and stress, which may be the neurophysiological basis for prolonged emotional stress inducing hypertension. The central amygdaloid nucleus (AC) is the most important in this circuit, which widely connects with the other nuclei via its CRF (corticotropin releasing factor)-ergic and SP (substance P)-ergic projection fibers. There is another pressor system composed of the lateral septum (SL), habenula (HB), locus coeruleus (LC), and rostral ventrolateral medulla (RVL); muscarinic receptors are involved in each connection of this system. In view of the facts that the SL also plays an important role in integration of emotion and autonomic reaction, and the AC projects to the SL, it is likely that the SL acetylcholine (ACh) pressor system is involved in the AC-emotional circuit. The present study demonstrates that injection of receptor blocker into each nucleus in the SL-ACh pressor pathway can reverse the AC pressor response, proving that the SL-HB (and HB-posterior hypothalamus)-LC-RVL pressor system is a component of the AC-emotional pressor circuit. PMID- 11911991 TI - Quantitation of polyamines in hypothalamus and pituitary of female and male developing rats. AB - The quantitation of four polyamines in hypothalamus and pituitary is studied in male and female developing rats using an improved high-performance liquid chromatography method. In the hypothalamus, putrescine (PUT) reaches the highest concentration (nmol/mg protein) on day 6. It shows the lowest value in comparison with any other polyamine. Spermidine (SPD) is high during the first postnatal days. Spermine (SPM) fluctuates, and agmatine (AGM) is highest during the first week. SPD, SPM and AGM are lower in females. In the pituitary, PUT, SPD and AGM are high during the first week. SPM remains constant and it is higher in males. AGM is higher in males only on day 1. PUT shows the lowest concentration of all. Concentrations of PUT, SPD and SPM are higher in the pituitary; AGM is higher in the hypothalamus. alpha-Difluoromethylornithine (a specific and irreversible inhibitor of ornithine decarboxylase) decreases PUT and SPD, increased SPM and AGM remain unchanged in the hypothalamus and pituitary. Thus, each polyamine has its own pattern in hypothalamus and in pituitary during development in males and females; these changes could be related to the hypothalamic control of pituitary secretion of hormones related to reproduction in mammals. PMID- 11911992 TI - Neuropeptide Y inhibits the hyperexcitability of type A neurons in chronically compressed dorsal root ganglion of the rat. AB - Our recent data revealed adrenergic sensitivity in chronically compressed dorsal root ganglion (DRG) of rats. As neuropeptide Y (NPY) is a common sympathetic co transmitter, we investigated the effect of NPY on injured DRG neurons. The expression of NPY Y1 and Y2 receptors and the effect of NPY on chronically compressed DRG neurons were studied using in situ hybridization and extracellular single fiber recording in vitro, respectively. After DRG compression, the expression of Y1 receptor was distinctly increased in large and medium-sized DRG neurons, while Y2 receptor was increased in small DRG neurons. NPY inhibited both the spontaneous activity and the excitatory effect of norepinephrine in injured DRG A-neurons. The results suggest a possibility that NPY may inhibit the hyperexcitability of injured DRG A-neurons via increased Y1 receptor following chronic compression. PMID- 11911993 TI - Increased production of nitric oxide stimulated by interferon-gamma from peripheral blood monocytes in patients with complex regional pain syndrome. AB - This study examines immediate nitric oxide (NO) release from monocytes following interleukin-1beta (IL-1beta), interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) challenge in patients with complex regional pain syndrome (CRPS). Study patients exhibited the following: (1), mechanical allodynia; (2), evidence of either vasomotor or sudomotor disturbance; and (3), concordant painful allodynia documented with quantitative sensory testing that was temporarily abolished with sympathetic block. Ten subjects (CRPS, N=5; control, N=5) were enrolled. Peripheral blood monocytes were challenged with 100 microl of IL-1beta (1 ng), IFN-gamma (1 ng), TNF-alpha (0.01 ng), and normal saline (NS) and the resultant immediate NO release measured. Subjects with CRPS exhibited a statistically significant increase in NO release in response to IFN gamma (P<0.012) compared with controls. The NO responses to IFN-gamma in excess of NS (P<0.025) and as the ratio IFN-gamma/NS (P<0.022) were also significantly increased. PMID- 11911994 TI - The addition of glyceryltrinitrate to capsaicin cream reduces the thermal allodynia associated with the application of capsaicin alone in humans. AB - The aim of this study was to determine whether topical application of capsaicin cream causes thermal allodynia and the extent to which this is attenuated by the addition of glyceryltrinitrate (GTN). This was a double blind placebo controlled study of 40 consenting adult subjects. Each of four cream combinations (GTN, capsaicin, GTN/capsaicin and vehicle) were applied to the subjects with at least a 1 day interval between each application. Water at a known temperature was applied to the standard area of skin where cream had been applied. Subjects rated the resulting thermal allodynia using a 0-10 Likhert score. Thermal allodynia is usually apparent when warm water is applied to skin containing capsaicin. The thermal allodynia caused by the topical application of capsaicin was significantly reduced by the addition of GTN. The addition of GTN to capsaicin cream significantly reduces the thermal allodynia associated with the application of capsaicin cream alone. PMID- 11911995 TI - Interleukin-1A polymorphism is not associated with late onset Alzheimer's disease. AB - Over the past few years, association studies have proposed a number of potential genetic risk factors for Alzheimer's disease (AD). With the exception of the varepsilon4 allele of the apolipoprotein E gene, whose association with the late onset type of AD (LOAD) has been confirmed, the relative significance of most of these associations is still in question. A polymorphism in the interleukin-1A gene (IL-1A2) has been suggested as a risk factor for the early onset as well as for LOAD. In this study, the distribution of IL-1A alleles was examined in a cohort of predominantly LOAD patients and in control individuals. No significant difference was detected in genotype or allele frequencies (odds ratios of 0.929 and 0.743, respectively; P>0.5). We conclude that IL-1A genotype is not a major risk factor for LOAD. PMID- 11911996 TI - Neuroactive steroid changes in response to challenge with the panicogenic agent pentagastrin. AB - BACKGROUND: Female hormones and female hormone derivatives, including neuroactive steroids (NASs) have been suspected to play a role in the pathophysiology of panic disorder (PD). The panicogenic agent CO(2) has been shown to induce a delayed release of NASs in both brain and plasma of rats. In the present study, we measured NASs plasma levels in response to challenge with another panicogenic agent, pentagastrin, and assessed the effect of ethynil estradiol (EE) pretreatment. METHODS: A double-blind cross-over placebo-controlled design with randomization of the order of a three day pretreatment of EE (50 microg/day) or placebo was used to assess the effect of a 30 microg iv bolus injection of pentagastrin on the release of allopregnanolone (ALLO) and dehydroepiandrosterone (DHEA) into plasma in 15 male PD patients and 10 male healthy volunteers (HV). RESULTS: After pentagastrin challenge there was a significant release of DHEA and a trend for the release of ALLO. EE pretreatment did not affect the pentagastrin induced panic response or NAS release. CONCLUSIONS: Pentagastrin induced release of NASs into plasma, the purpose of which remains to be determined. PMID- 11911997 TI - Effects of oral estrogen, raloxifene and arzoxifene on gene expression in serotonin neurons of macaques. AB - The serotonin neural system contributes to cognition and affect, both of which exhibit pathologies with gender bias. We previously showed that estrogen (E) treatment of female macaques via Silastic implant alters gene expression for tryptophan hydroxylase (TPH), the serotonin reuptake transporter (SERT) and the 5HT1A autoreceptor. In addition, we have found that serotonin neurons of macaques express ER beta (ER beta). Together these studies suggest that the serotonin neural system could transduce the action of estrogen via ER beta on aspects of mood and cognition. However, estrogen replacement therapy can increase the risk for breast and uterine cancer. Therefore, we questioned whether the selective estrogen receptor modulators, raloxifene and arzoxifene, act in a manner similar to E on gene expression in serotonin neurons of a nonhuman primate model. Female rhesus macaques were ovariectomized and orally dosed with vehicle, estradiol 17beta, raloxifene or arzoxifene once per day by sipper bottles for 30 days. The animals were then euthanized and the midbrains were prepared for in situ hybridization for TPH, SERT and 5HT1A receptor mRNAs followed by densitometric analysis. There was a significant increase in TPH total signal (positive pixelsxOD) with E, raloxifene and arzoxifene, respectively. There was a significant decrease in SERT mRNA optical density with all treatments. 5HT1A autoreceptor mRNA did not change with any treatment. If these changes in gene expression are reflected by similar changes in the functional proteins, then raloxifene or arzoxifene could increase serotonin neurotransmission with little or no negative action in peripheral tissues. In conclusion, the selective estrogen receptor modulators, raloxifene and arzoxifene, act in a manner similar to natural E on TPH and SERT mRNA expression in serotonin neurons. This suggests that raloxifene and arzoxifene are agonists at ER beta in the context of the serotonin neuron. However, the responses to E were more variable and less robust with the oral dosing paradigm compared to a chronic implant paradigm. PMID- 11911998 TI - Mental deterioration correlates with response of natural killer (NK) cell activity to physiological modifiers in patients with short history of Alzheimer's disease. AB - Natural killer (NK) cell activity of peripheral blood mononuclear (PBM) cells was measured in 16 subjects with mild to moderate senile dementia of Alzheimer's type (sDAT) chosen for short history of disease and no medication, and in 17 age- and sex-matched controls. Levels of cytotoxicity at baseline and after PBM cell exposure to modifiers either negative (cortisol 10(-6) M) or positive (rIL-2 650 IU/ml and rIFN-gamma 100 UI/ml, respectively) were related to indices of hypothalamic-pituitary-adrenal (HPA) function and Gottfries Brane Rating Scale (GBS) score for mental deterioration. Spontaneous NK cell activity was not significantly different in sDAT subjects vs controls. In vitro inhibition by cortisol was lower in sDAT (P<0.05); cytokine-induced changes were greater (rIL 2, P<0.02; rIFN-gamma, P<0.05). Percent negative or positive variations from baseline significantly correlated with GBS scores (P<0.05 or less). Serum cortisol and cortisol/DHEAS molar ratio at 0800 h were significantly higher in sDAT (P<0.05 and P<0.02, respectively). Cortisol/DHEA ratio positively correlated with GBS scores (P<0.02). Moreover, the ratios of incremental area of response ACTH/cortisol and beta-endorphin/cortisol after 1 microg/kg ovine-corticotrophin releasing hormone (o-CRH) positively correlated with percent increase of NK cell activity after rIL-2 (P<0.01). Data indicate that patients with mild cognitive impairment and short history of sDAT show abnormal responsiveness of NK cell activity to physiological modifiers while maintaining normal spontaneous activity. Furthermore, data are compatible with partial glucocorticoid resistance at the immune level. Progressing sDAT longitudinal studies are needed to address: i) the clinical applicability of these abnormalities as prognostic factors; ii) the role played by pro-opiomelanocortin (POMC)-derived peptides and adrenal androgens in the control of NK cell activity. PMID- 11911999 TI - Lack of seasonal rhythms in central serotonergic function in boys with ADHD. AB - This study examined seasonal variations in central 5-HT function in a relatively large sample of prepubertal boys with attention-deficit/hyperactivity disorder (ADHD). In addition, seasonal differences in central 5-HT function among aggressive and non-aggressive children in the sample were explored. Ninety-three boys with ADHD were divided into aggressive (n=54) and nonaggressive (n=39) groups based on parental responses to interviews and ratings of behavior. Central 5-HT function was assessed by measuring the prolactin response to a single 1 mg/kg oral dose of d,l-fenfluramine. The prolactin values were derived from different children over the course of eight years, but were collapsed across years and analyzed in terms of day of the year. Cosinor analysis revealed no annual, bi-annual, 3-monthly, 2-monthly, monthly, or bi-monthly rhythms in the prolactin response in the entire sample of boys with ADHD. Moreover, there was no evidence of seasonal differences in the prolactin response between the aggressive and non-aggressive subgroups. These results indicate that seasonality-related variance is not a threat to the validity of the neurochemical procedure and does not account for the inconsistencies in the studies of central 5-HT and aggression in children with ADHD. PMID- 11912000 TI - Vasoconstrictor response to topical beclomethasone in major depression. AB - Overactivity of the hypothalamic-pituitary-adrenal (HPA) axis has been frequently described in depression. Due to the closed-loop nature of the HPA axis, one possible cause of this overactivity may be a defect in negative feedback regulation, in particular an abnormality of the glucocorticoid receptor (GR). In the present study, the vasoconstrictor response to the topical glucocorticoid, beclomethasone, was used to examine GR function in depression. Topical beclomethasone was applied in four concentrations (10 microl each of 3, 10, 30 and 100 microg/ml) to the forearms of 22 subjects with major depression and their age- and sex-matched controls. Skin blanching responses were compared between the depressed and control groups and, within the depressed group, on the basis of the modified dexamethasone suppression test (DST), between cortisol suppressors and non-suppressors. Depressed subjects demonstrated a significantly reduced vasoconstrictor response compared to controls (P=0.0001). No difference was detected between cortisol suppressors and non-suppressors in their skin blanching responses. These findings suggest that peripheral GR function is abnormal in depression but that the reduced vasoconstrictor response to beclomethasone is not necessarily a secondary effect of hypercortisolaemia or HPA axis overactivity. PMID- 11912001 TI - Sexual behavior in lesbian and heterosexual women: relations with menstrual cycle phase and partner availability. AB - Using a prospective design over three complete menstrual cycles, 147 heterosexual and 89 lesbian women made daily recordings of their basal body temperature (BBT), cervical mucus status, menses, and completed a daily checklist of various sexual behaviors (including sexual self-stimulation and sexual activity with a partner). They also gave their age, height, weight, age at menarche, number of pregnancies, duration of sleep, tobacco, caffeine, and alcohol use, and whether they had a live-in sexual partner. Using BBT, cervical mucus status, and menses information, cycle days were grouped into five discrete phases: menses, follicular, ovulatory, early luteal, and premenstrual. Daily frequencies of sexual behavior with a partner and autosexual behavior were computed for each phase. Mixed ANOVAs on the resultant proportional data revealed similar patterns for autosexual behavior across the phases for both heterosexuals and lesbians who did not have a live-in partner, in which autosexual behavior was highest during the follicular and ovulatory phases. For those with live-in partners, autosexual behavior did not vary across the phases. Lesbians engaged in more autosexual behavior overall. Allosexual behavior peaked during the follicular phase for both heterosexuals and lesbians, and the phasic pattern was unrelated to live-in partner status. Additional analyses suggest that the observed patterns were unrelated to anticipated changes in sexual activity due to menses. Results are discussed in terms of social variables and hormonal fluctuations associated with the menstrual cycle. PMID- 11912003 TI - Menstrual synchrony and cycle variability: a reply to Schank (2000). AB - Menstrual synchrony has been measured by a change over time, to capture the "process" of synchronization or by examining a sample of women who have been together for a number of months ("state" synchrony). We have proposed and used a method for calculating "state" synchrony. On the basis of the results of a computer simulation, Schank (Schank, J.C., 2000. Menstrual-cycle variability and measurement: further cause for doubt. Psychoneuroendocrinology 25, 837-847) argues that our method is biased towards finding synchrony and that the degree of error is an increasing function of cycle variability. We show that Schank's model is based on unfounded assumptions. Contrary to Schank's assertion, on which much of the critique is based, there is no evidence that cycle variability is actually large in samples with "state" synchrony. Our reports are valid; and our method is supported both by convergent validity and by the use of statistical "random control" groups. PMID- 11912002 TI - Cognitive impairment correlates with hypothalamo-pituitary-adrenal axis dysregulation in multiple sclerosis. AB - Hypothalamo-pituitary-adrenal (HPA) dysregulation has recently been demonstrated in multiple sclerosis (MS) by means of combined dexamethasone corticotropin releasing hormone (Dex-CRH) suppression tests. Authors found a correlation with course of disease and to a lesser extent with depressive symptoms. In this study, we aimed to further evaluate whether HPA disturbances in MS are correlated with cognitive impairment, disability status, and fatigue. Dex-CRH tests were performed in a total of 40 patients and 11 healthy controls. Concomitantly, cognitive impairment was evaluated using the symbol digit modalities test and fatigue was assessed by different fatigue severity scales. When comparing patient subpopulations to healthy subjects, Dex-CRH stimulation tests indicated an HPA hyperactivity in primary and secondary progressive MS, while relapsing-remitting patients had response patterns similar to controls. However, results were only significant for one of the six analysed parameters, i.e. area under the curve calculations of ACTH stimulation. Within the patient sample, clear-cut differences emerged between groups of different cognitive impairment, being significant for all ACTH response parameters. Our results suggest an HPA hyperactivation related to increased cognitive impairment. Indicators of HPA axis activation further correlated substantially with neurologic disability, but only moderately with duration of disease and even less with depressive symptoms and fatigue. We conclude that the observed dysregulation is more likely a secondary effect of the extent of brain damage rather than primarily involved in the pathogenesis of MS. PMID- 11912004 TI - The stem cell compartment in human interfollicular epidermis. AB - In this review, I summarise recent work from my laboratory in which we have been examining the distribution of stem cells in human interfollicular epidermis and the factors that regulate stem cell fate in vitro. The non-random distribution of stem cells is emphasised and beta1 integrins and Delta1 are suggested to play a role in stem cell patterning. beta1 integrins, Notch, c-Myc and beta-catenin all regulate the size of the stem cell compartment in vitro and recent evidence from transgenic mice suggests that they are also important in vivo. PMID- 11912005 TI - A novel homozygous point mutation in the COL17A1 gene in a Chinese family with generalized atrophic benign epidermolysis bullosa. AB - We describe a Chinese family with generalized atrophic benign epidermolysis bullosa (GABEB), a non-lethal variant of junctional epidermolysis bullosa. The proband was an offspring of consanguineous parents, had generalized blisters since birth and developed severe alopecia during early childhood. Ultrastructural examination of the proband's skin revealed fissures in the lamina lucida. Immunofluorescence assays using a monoclonal antibody recognizing the extracellular domain of the 180 kDa bullous pemphigoid antigen (BPAG2) showed loss of fluorescent signal in the basal membrane zone of the skin. DNA sequencing revealed a homozygous C-to-G transversion at nucleotide position 899 in exon 11 of the COL17A1 gene, which encodes BPAG2. This mutation results in serine to cysteine at position 265, which is located in a highly conserved region of the intracellular domain of BPAG2. We showed that the proband's father was heterozygous for this mutation. In addition, we found a novel polymorphic substitution of C-to-G at nucleotide position 798 in exon 10 of the COL17A1 gene, which results in an I233M change in BPAG2 and is a common polymorphic allele in a limited Chinese population. PMID- 11912006 TI - Aberrant expression of apoptosis-related molecules in psoriatic epidermis. AB - Apoptosis is a physiological form of cell death that is responsible for the deletion of cells. Epidermal keratinocytes are supposed to be regulated by cell proliferation and cell death leading to structural homeostasis. Psoriatic skin shows marked thickening of the epidermis, suggesting the imbalance of the homeostasis, which might be related to abnormal apoptotic process. We investigated the expression of various apoptosis-related molecules in the psoriatic hyperproliferative epidermis. Real time quantitative RT-PCR analyses revealed that mRNAs of Fas, Bcl-xL, Bax and ICAD (inhibitor of caspase 3-related DNase) of the psoriatic involved epidermis were increased by 4.2-, 2.8-, 2.6- and 5.6-fold, respectively, compared with the uninvolved epidermis. In contrast, Bcl 2 expression in the involved epidermis was one-third suppressed compared with the uninvolved epidermis. No significant difference in the expression of mRNAs of Fas ligand or CAD (caspase 3-related DNase) was detected between the involved and uninvolved epidermis. Western blot analysis and immunohistochemical studies showed compatible results obtained by RT-PCR analyses. Although active caspase 3 was slightly increased in the involved epidermis, apoptotic cells were marginally detected. These results indicate that psoriatic epidermis shows aberrant expression of apoptosis-related molecules representing suppressed apoptotic process, which might be related to characteristic histopathology. PMID- 11912007 TI - Unique therapeutic effects of the Japanese-Chinese herbal medicine, Sairei-to, on Th1/Th2 cytokines balance of the autoimmunity of MRL/lpr mice. AB - Sairei-to, one of the Japanese-Chinese herbal medicines has been used for the treatment of various diseases, especially collagen disease and edema in nephrotic syndrome. However, the mechanism of the therapeutic effects remains uncertain. Therefore, we investigated the immunological changes of skin, kidney, spleen cells and serum in autoimmune-prone MRL/lpr, MRL/n and C57BL/6J mice treated with Sairei-to. In MRL/lpr mice treated with Sairei-to, the improvement of proteinuria, reduction in the number of hematoxylin bodies in kidney, and reduced serum levels of blood urea nitrogen were observed. These results indicate that Sairei-to can improve or inhibit the progression of lupus nephritis. The proportion of CD19 and the serum levels of IgG1, which is one of the pathogenesis of lupus dermatoses and lupus nephritis, were significantly reduced in Sairei-to treated MRL/lpr mice. Therefore, it is suspected that the B cell function was suppressed by Sairei-to. In addition, CD4/8 ratio in spleen cells and the degree of lymphoproliferation in MRL/lpr mice also decreased. Interestingly, IL-4 producing spleen cells were increased significantly by ELISPOT assay, and IFN gamma mRNA expressions were reduced in Sairei-to-treated MRL/lpr mice. Regarding the Th balance, an imbalance towards Th1 predominance may play a significant role in MRL/lpr mice, and the Th1 axis was suppressed and the Th2 axis became predominant in Sairei-to-treated MRL/lpr mice. On the other hand, Th2 cell type immunoglobulins (IgG1) were suppressed. These results suggested that Sairei-to is potential for impairing shifted Th1/Th2 balance and hypergammaglobulinemia resulting in therapeutic effects. PMID- 11912008 TI - New formulation of chemical peeling agent: 30% salicylic acid in polyethylene glycol. Absorption and distribution of 14C-salicylic acid in polyethylene glycol applied topically to skin of hairless mice. AB - Salicylic acid is used in chemical peeling procedures. However, they have caused many side effects, even salicylism. To achieve a salicylic acid peeling that would be safer for topical use, we recently developed a new formulation consisting of 30% salicylic acid in polyethylene glycol (PEG) vehicle. In an extension of our previous research, we studied the absorption of 30% salicylic acid labeled with 14C in PEG vehicle applied topically to the intact and damaged skin of male hairless mice. An ointment containing 3 mg salicylic acid in 10 mg vehicle was applied to both groups. In animals with intact skin, 1 h after application the plasma concentration of radioactivity was 1665.1 ng eq/ml, significantly lower than the 21437.6 ng eq/ml observed in mice with damaged skin. Microautoradiograms of intact skin showed that the level of radioactivity in the cornified cell layer was similar at 6 h after application. However, in damaged skin, the overall level of radioactivity showed a decrease by 3 h after application. In the carcasses remaining after the treated intact and damaged skin had been removed, 0.09 and 11.38% of the applied radioactivity remained, respectively. These findings confirm that 30% salicylic acid in PEG vehicle is little absorbed through the intact skin of hairless mice, and suggest that salicylism related to absorption through the skin of quantities of topically applied salicylic acid is not likely to occur in humans with intact skin during chemical peeling with this preparation. This new preparation of 30% salicylic acid in PEG vehicle is believed to be safe for application as a chemical peeling agent. PMID- 11912009 TI - Lack of involvement of P-glycoprotein (P-gp) in pemphigus patients with poor response to steroid therapy. AB - A small but significant fraction of pemphigus patients do not show an adequate response to steroid therapy. P-glycoprotein (or P-gp) is a cell membrane efflux pump that expels drugs, including glucocorticoids, from the cytosol to the extracellular medium. An increased expression and/or function of P-glycoprotein in lymphoid cells could decrease the intracellular concentration of glucocorticoids, diminishing its therapeutic effects. The aim of this work was to assess the expression and activity of P-glycoprotein in the mononuclear cells (MNC) from pemphigus patients with good and poor response to steroid therapy. We studied 20 patients with pemphigus vulgaris, eight of them classified as poor responders and 12 as good responders to steroid therapy. The expression and activity of P-glycoprotein by MNC were quantified by flow cytometry, and P glycoprotein mRNA levels were determined by a semi-quantitative RT-PCR technique. We found that the expression of P-glycoprotein at both mRNA and protein levels was similar in pemphigus patients with good and poor response to steroid therapy. Similar results were obtained regarding P-glycoprotein activity. P-glycoprotein does not seem to be involved in the poor response to steroid treatment seen in some pemphigus patients. It is important to investigate additional mechanisms that could account for the glucocorticoid resistance seen in some pemphigus patients. PMID- 11912010 TI - Overactivation of IL-4-induced activator protein-1 in atopic dermatitis. AB - Atopic dermatitis is regarded as mediated by Th2-type immunity. In fact, it frequently coincides with the elevation of immunoglobulin (Ig)-E in patients' sera. Due to the pivotal role of interleukin (IL)-4 in regulation of IgE, we hypothesized if atopic dermatitis represents a hyper-reactive condition in response to IL-4 when it coincides the higher serum level of IgE. To address this possibility, peripheral blood mononuclear cells (PBMC) isolated from patients with atopic dermatitis with the high serum IgE level, from those with psoriasis or from healthy volunteers were stimulated with recombinant IL-4 and analyzed for activation of transcription factors including activator protein (AP)-1 or signal transducers and activators of transcription (STAT)-6 by employing electrophoretic mobility shift assays. Although no significant difference between atopy patients and other groups was observed in the STAT-6 binding activity in IL-4-stimulated PBMC, it over-activated the binding of AP-1 in PBMC of the patients with atopic dermatitis. The AP-1 binding was interfered by the use of an antibody directed against JunB. This is the indication that IL-4-overactivated AP-1 is composed of JunB. Furthermore, semi-quantitative RT-PCR analyses revealed marked down modulation of a Th1 cytokine, interferon (IFN)-gamma, in IL-4-stimulated PBMC derived from atopy patients, but not that from healthy individuals. Together, our present study indicates that AP-1 is over-activated by IL-4 in PBMC of the atopic patients with the higher IgE level, thereby implying that IL-4-induced over activation of AP-1 might be one of pathogenic factors in atopic dermatitis. PMID- 11912011 TI - Dominant expression of CXCR3 is associated with induced expression of IP-10 at hapten-challenged sites of murine contact hypersensitivity: a possible role for interferon-gamma-producing CD8(+) T cells in IP-10 expression. AB - Murine contact hypersensitivity is elicited as a consequence of immunologic reactions triggered by skin-applied antigen, interactions among Langerhans cells, T cells, keratinocytes and mast cells, and a variety of chemokines generated by cellular interactions. In this study, we sensitized and challenged BALB/c mice with hapten, dinitrofluorobenzene or picryl chloride, and examined the expressions of mRNA for chemokines and their receptors by reverse transcription polymerase chain reaction in the skin of elicited earlobes. CXC chemokines, IP-10 and Mig, were transcribed 24-48 h after challenge. This was associated with the expression of their agonistic receptor CXCR3, while mRNA for TARC and MDC, and their receptor CCR4 were not detected. Since CXCR3 and CCR4 are expressed preferentially on types 1 (Th1/Tc1) and 2 (Th2) T cells, respectively, the results suggested that the former type of T cells predominantly infiltrate at the elicited sites. Immune lymph node cells of the sensitized mice also expressed mRNA for CXCR3 but not CCR4 with concomitant transcription of interferon-gamma (IFN-gamma) but not interleukin-4 subsequent to challenge. The percentage of lymph node CD8(+) T cells was increased from 16% in naive mice to 30-50% in hapten-challenged mice, and in the immune lymph nodes, CD8(+) cells were the major source of IFN-gamma compared to CD4(+) cells. Since IFN-gamma is known to stimulate keratinocytes to produce IP-10 and Mig, it is suggested that these IFN gamma-producing CD8(+) T cells enhance the production of these chemokines, thereby functioning as not only the effector cells but also the cytokine source to sustain the challenge reaction. PMID- 11912012 TI - Abstracts of the 7th ECNP (European College of Neuropsychopharmacology) Regional Meeting. Bucharest, Romania, April 19-21, 2002. PMID- 11912013 TI - Exploiting genome sequence: predictions for mechanisms of Campylobacter chemotaxis. AB - The genome sequence of Campylobacter jejuni NCTC 11168 reveals the presence of orthologues of the chemotaxis genes cheA, cheW, cheV, cheY, cheR and cheB, ten chemoreceptor genes and two aerotaxis genes. The presence of cheV and a response regulator domain in CheA, combined with the absence of a cheZ gene and the lack of a response regulator domain in CheB, reveals significant differences in the C. jejuni chemotaxis system compared with that found in other bacteria. PMID- 11912016 TI - Armed to the teeth. PMID- 11912014 TI - Natural transformation in Helicobacter pylori: DNA transport in an unexpected way. AB - Like other bacterial species with a high frequency of inter-strain recombination, the human gastric pathogen Helicobacter pylori is competent for natural transformation. Recent data, however, indicate that its DNA-uptake system differs significantly from that in other species that contain DNA-uptake systems related to type IV pili. Instead, in H. pylori it has been suggested that the five proteins that form the transmembrane channel of the transformation system are closely related to subunits of type IV secretion systems. PMID- 11912017 TI - SHP-2 tyrosine phosphatase--the target of CagA. PMID- 11912020 TI - Strain families of Mycobacterium tuberculosis. PMID- 11912022 TI - The Schaechter-Bentzon-Maaloe experiment and the analysis of cell cycle events in eukaryotic cells. AB - The Schaechter-Bentzon-Maaloe (SBM) experiment, performed more than 40 years ago, provides an important lesson for the analysis of the eukaryotic cell cycle. Before this experiment, temperature shifts had been used to synchronize bacteria and determine the pattern of DNA synthesis during the bacterial division cycle. These experiments indicated that DNA replication occurred during a fraction of the division cycle with gaps before and after DNA synthesis, a pattern similar to the eukaryotic division cycle. The SBM experiment studied DNA replication during the division cycle by labeling an unperturbed culture with a short pulse of tritiated thymidine. All cells were found to be labeled, indicating that unperturbed cells synthesize DNA throughout the division cycle. Thus, the SBM experiment was a control experiment demonstrating that artifacts can be introduced by synchronization methods. The idea of an control experiment under unperturbed conditions is proposed for the analysis of data on cell-cycle specific gene expression in yeast and mammalian cells. PMID- 11912023 TI - Anti-fungal therapy at the HAART of viral therapy. AB - HIV-positive patients receiving combination therapy (highly active anti retroviral treatment, HAART) suffer significantly fewer oral infections with the opportunistic fungal pathogen Candida albicans than non-HAART-treated patients. One component of HAART is an inhibitor of the HIV proteinase, the enzyme required for correct processing of retroviral precursor proteins. It would appear that HIV proteinase inhibitors also have a direct effect on one of the key virulence factors of C. albicans, the secreted aspartic proteinases (Saps). This suggests that the reduction in C. albicans infections in HIV-positive patients might not be solely the result of improved immunological status but could also be caused by the HAART treatment directly inhibiting Candida proteinases. PMID- 11912024 TI - HIV proteinase inhibitors: do they really work against Candida in a clinical setting? PMID- 11912025 TI - How do bacteria resist human antimicrobial peptides? AB - Cationic antimicrobial peptides (CAMPs), such as defensins, cathelicidins and thrombocidins, are an important human defense mechanism, protecting skin and epithelia against invading microorganisms and assisting neutrophils and platelets. Staphylococcus aureus, Salmonella enterica and other bacterial pathogens have evolved countermeasures to limit the effectiveness of CAMPs, including the repulsion of CAMPs by reducing the net negative charge of the bacterial cell envelope through covalent modification of anionic molecules (e.g. teichoic acids, phospholipids and lipid A); expelling CAMPs through energy dependent pumps; altering membrane fluidity; and cleaving CAMPs with proteases. Mutants susceptible to CAMPs are more efficiently inactivated by phagocytes and are virulence-attenuated, indicating that CAMP resistance plays a key role in bacterial infections. PMID- 11912026 TI - Port of entry--the type III secretion translocon. AB - Many Gram-negative plant and animal pathogenic bacteria use a specialized type III secretion system (TTSS) as a molecular syringe to inject effector proteins directly into the host cell. Protein translocation across the eukaryotic host cell membrane is presumably mediated by a bacterial translocon. The structure of this predicted transmembrane complex and the mechanism of transport are far from being understood. In bacterial pathogens of animals, several putative type III secretion translocon proteins (TTPs) have been identified. Interestingly, TTP sequences are not conserved among different bacterial species, however, there are structural similarities such as transmembrane segments and coiled-coil regions. Accumulating evidence suggests that TTPs are components of oligomeric protein channels that are inserted into the host cell membrane by the TTSS. PMID- 11912030 TI - From synaptic errors to thalamocortical circuitry. AB - Recent data indicate that newly grown synapses in the brain are not guaranteed to innervate their desired target, but can form instead on nearby targets. Such 'errors' introduce representational inaccuracies but improve representational flexibility. Optimizing accuracy and flexibility requires detecting correlated activity and disabling plasticity, explaining the structure of the thalamocortical circuit. PMID- 11912027 TI - Intracellular vs extracellular recognition of pathogens--common concepts in mammals and flies. AB - There are common themes in innate immune defense systems across the animal and plant kingdoms. Pathogen recognition is commonly based on the identification of microbial molecular patterns by defined receptors and the subsequent activation of signaling pathways that initiate a defense response to fend off the invading microorganism. The existence of mammalian Toll-like receptors (TLRs) and the recent identification of two mammalian nucleotide-binding site leucine-rich repeat (NBS-LRR) proteins (NOD1 and NOD2) as intracellular sensors of bacterial products bring new insights into the possibility of extracellular versus intracellular pathogen recognition and signal transduction depending on the nature of the infection. The homology between TLRs and the Toll system in Drosophila suggests that conserved defense mechanisms are likely to be shared by diverse organisms. PMID- 11912031 TI - Fixing unsaid meanings. AB - The First International Workshop on Semantics, Pragmatics and Rhetorics (SPR-01) was held in Donostia-San Sebastian, Spain, on 22-24 November 2001. The conference was organized by the Institute for Logic, Cognition, Language and Information (ILCLI), and the Department of Logic and Philosophy of Science of the University of the Basque Country (UPV/EHU). PMID- 11912032 TI - Plumbing semantic depths in Amsterdam. AB - The 13th Amsterdam Colloquium was held at the Institute for Logic, Language and Linguistics at the University of Amsterdam, on 17-19 December 2001. PMID- 11912034 TI - Left hemisphere discourse? PMID- 11912033 TI - Seeing patterns in human visual cortex. PMID- 11912035 TI - Probing perceptual asynchrony. PMID- 11912037 TI - Social cognition and primacy of movement revisited. PMID- 11912038 TI - Motion integration during motion aftereffects. AB - The perceived global motion of a stimulus depends on how its different local motion-direction vectors are distributed in space and time. When they are explicitly co-localized, as in the case of locally paired motion, competitive motion integration mechanisms produce a unitary global motion direction determined by their vector average. During motion aftereffects induced by simultaneous adaptation to multiple motion directions, just as in the case of locally paired motion, different directional signals originate simultaneously from exactly the same position in space. Therefore, the perceived global motion direction during motion aftereffects results from local vector averaging of the co-localized motion-direction signals induced by adaptation. PMID- 11912039 TI - Episodic memory and cortico-hippocampal interactions. AB - There is a broad consensus that the hippocampal system plays a critical role in the encoding and retrieval of 'episodic' memories. Recent findings and computational modeling explicate the representational requirements of encoding episodic memories, and suggest that the idiosyncratic architecture of the hippocampal system and its interactions with cortical circuits are well-matched to the representational problems it must solve in order to support the episodic memory function. These findings also shed light on the nature of consolidation, identify the sorts of memories that must remain encoded in the hippocampal system for the long-term, and help delineate the semantic and episodic memory distinction. PMID- 11912040 TI - Genetics and general cognitive ability (g). AB - Two recent articles in this journal made the case for the existence and importance of g and reviewed research on cognitive and psychophysical correlates of psychometric g. This review considers g from a genetic perspective. Multivariate genetic research indicates that g accounts for nearly all of the genetic variance of diverse psychometric cognitive tests (genetic g). Recent research suggests not only that elementary cognitive tasks are genetically linked to psychometric g but also that genetic g pervades these tasks. Contrary to the assumption of modularity that dominates cognitive science, genetic g exists in the mind as well as in psychometric tests. PMID- 11912041 TI - The topography of high-order human object areas. AB - Cortical topography is one of the most fundamental organizing principles of cortical areas. One such topography - eccentricity mapping - is present even in high-order, ventral stream visual areas. Within these areas, different object categories have specific eccentricity biases. In particular, faces, letters and words appear to be associated with central visual-field bias, whereas buildings are associated with a peripheral one. We propose that resolution needs are an important factor in organizing object representations: objects whose recognition depends on analysis of fine detail will be associated with central-biased representations, whereas objects whose recognition entails large-scale integration will be more peripherally biased. PMID- 11912042 TI - Sperm control. AB - Evidence is accumulating that in many species, males deliver more sperm to some females and fewer to others depending on how valuable they perceive the copulation. Do humans have any control over the number of sperm they ejaculate? PMID- 11912043 TI - On what's left and still right about assymetry. PMID- 11912045 TI - Environmental stressors - Pollutants symposium. Proceedings of the 21st International Congress of the European Society of COmparative Physiology and Biochemistry (ESCPB). Liege, Belgium. July 24-28, 2000. PMID- 11912044 TI - Madness in the garden or worm in the apple? PMID- 11912046 TI - Cloning and sequencing of a novel metallothionein gene in Mytilus galloprovincialis Lam. AB - Metallothionein (MT) is a ubiquitous, metal-inducible protein with an important role in the homeostasis and in the detoxification of heavy metals. This work reports the cloning and sequencing of a MT gene encoding a MT isoform (MT20-IIIa) in the mussel Mytilus galloprovincialis Lam, a lamellibranch mollusc known to accumulate and to detoxify large amounts of metal. The MT gene, lacking the 5' promoter region, is 1865 bp long and has a tripartite structure consisting of three exons and two introns. The putative open reading frame (ORF) encodes a polypeptide of 72 amino acids, which corresponds to the MT-I class, type 2 family (http://www.unizh.ch/~mtpage/classif.html). The structure of the gene and the putative MT20-III protein have been compared with those of other species. The putative biological significance of the differences at the amino acid level among the different MTs is discussed. PMID- 11912047 TI - Transport of cadmium across the apical membrane of epithelial cell lines. AB - Cadmium (Cd) uptake and secretion across the apical membrane of epithelial cells was studied using LLC-PK1 cells cultured on Petri dishes and permeable membranes, respectively. Cd accumulation in cells from the apical medium was decreased by low temperature and metabolic inhibitors. A saturable tendency was observed between initial Cd accumulation and increased concentrations of Cd in the apical medium at 37 degrees C, but not at 4 degrees C. Co-incubation with ZnCl2 or CuCl2 competitively decreased Cd accumulation at 37 degrees C. A decrease in the pH of the apical medium markedly decreased Cd accumulation. Pretreatment of cells with an inorganic anion-exchange inhibitor significantly decreased Cd uptake at pH 7.4 in the presence of bicarbonate, but only marginally in its absence. A decrease in the pH of the apical medium increased the secretory (basolateral-to-apical) transport of Cd, with a concomitant decrease in the cellular accumulation of Cd. Co-incubation with Cd and tetraethylammonium, a typical substrate of the organic cation transporter, decreased Cd transport, with a concomitant increase in cellular Cd accumulation. The uptake and secretion of Cd across the apical membrane appear to be partly mediated via an inorganic anion exchanger and a H+ antiport of the organic cation transport system, respectively. Therefore, a decrease in pH of the apical medium markedly decreases Cd accumulation, possibly as a result of not only the decrease in Cd uptake via an inorganic anion exchanger, but also the increase in Cd secretion via the Cd2+/H+ antiport. Further evidence of the antiport was obtained from experiments using brush border membrane vesicles isolated from rat kidney and small intestine. In addition, passive diffusion of Cd appears to be decreased by low temperature and a decrease in pH. PMID- 11912048 TI - Alterations of tissue glutathione levels and metallothionein mRNA in rainbow trout during single and combined exposure to cadmium and zinc. AB - The objective of this study was to assess the effects of Cd and Zn exposure of rainbow trout (Oncorhynchus mykiss) on (a) hepatic glutathione (GSH) levels; and (b) hepatic and branchial metallothionein (MT) mRNA expression. Juvenile rainbow trout were exposed to waterborne Cd (nominal concentrations: 1.5 or 10 microg Cd l(-1)), Zn (150 or 1000 microg Zn l(-1)) or Cd/Zn mixtures (1.5 microg Cd l(-1) with 200 microg Zn l(-1) or 10 microg Cd l(-1) with 1000 microg Zn l(-1)). After 14 and 28 days of treatment, hepatic concentrations of total glutathione, oxidized glutathione (GSSG) and cysteine were determined by means of fluorometric high performance liquid chromatography (HPLC). Branchial and hepatic expression of MT mRNA was measured by means of semi-quantitative RT-PCR. Exposure of trout to Zn did not result in significantly elevated tissue levels of Zn, whereas Cd accumulation factors changed significantly with time and concentration. Despite of the absence of Zn accumulation, hepatic GSH but not MT mRNA levels were significantly altered in Zn-exposed fish. Cd, on the contrary, affected mainly the MT response but not GSH. Also tissue specific differences in the regulation of the two thiol pools were expressed. The thiol response after exposure to metal mixtures could not be explained by simple addition of the effects of the individual metals. The results indicate that cellular thiol pools show different reaction patterns with respect to specific metals and metal mixtures. Under conditions of long-term, low dose metal exposure, the function of GSH appears to go beyond that of a transitory, first line defense. PMID- 11912049 TI - White-sided dolphin metallothioneins: purification, characterisation and potential role. AB - Metallothioneins (MTs) were characterised in the kidneys of a white-sided dolphin Lagenorhynchus acutus stranded along the Belgian coast, displaying high levels of cadmium (Cd) and mercury (Hg) in liver and kidney. The protein has two isoforms: MT-1 and MT-2. MT-1 binds Cu, Zn, Hg and Cd, while MT-2 only binds Zn, Hg and Cd. This suggests different metabolic functions for the two isoforms: MT-1 is mainly involved in Cu homeostasis; MT-2, which was four-fold more abundant than MT-1, detoxifies most of the accumulated cadmium. PMID- 11912050 TI - Daily stress protein (hsp70) cycle in chitons (Acanthopleura granulata Gmelin, 1791) which inhabit the rocky intertidal shoreline in a tropical ecosystem. AB - Stress protein (heat shock protein, hsp70) response is involved in protecting organisms from the detrimental effects of environmental stressors, such as radiation and high temperatures. Tropical chitons can briefly tolerate high temperatures. However, they minimize the effects of elevated temperature during daylight hours and periods of tidal air exposure by remaining in rocky intertidal microhabitats along the shoreline of tropical waters. To study the natural variability of the hsp70 level, individuals of the polyplacophoran species Acanthopleura granulata Gmelin, 1791 were sampled every 4 h on two days in spring of 1999. Hsp70 levels were separately measured in the supernatant of the intestinal tract and foot muscle homogenates with a standardized immunoassay. The hsp70 level in the intestinal tract was highest in the early morning, decreased during the mid-morning hours and dropped to a comparatively low level in the afternoon, before increasing again during the night. The stress protein level in the foot muscle followed the daily air temperature curve with a time delay of a few hours, reaching the highest level in the afternoon and the lowest level in the early morning. The stress protein response can be interpreted as a sign of heat tolerance development and may play a role in allowing A. granulata to tolerate the temperature variability typical of its intertidal habitat. PMID- 11912051 TI - Mannosomes: a molluscan intracellular tubular membrane system related to heavy metal stress? AB - Amongst animals, several hydrogen peroxide-generating oxidases are apparently restricted to molluscs. One of these, D-mannitol oxidase, is concentrated in the alimentary system, where it is associated with its own subcellular membrane system of unique tubular morphology, most likely representing a structural modification of the ER. These structures can be purified by subcellular fractionation and have been termed 'mannosomes'. Little is known about the functions of mannitol oxidase or of mannosomes, but the previously reported molluscicide-induced increase in mannosomes implies their involvement in a general stress reaction. In this study, we examined the effects of heavy metal stress in the terrestrial gastropod Arion lusitanicus. The activity of mannitol oxidase and mannosome abundance were monitored, together with metal effects on heat-shock protein level, and these parameters were compared to heavy metal accumulation in the digestive gland. We found that mannitol oxidase is inhibited by heavy metals more than other oxidases. On the other hand, hsp70 levels and mannosomal protein were increased with enhanced heavy metal stress, the latter indicating a probable increase in the number of mannosome organelles. Thus, stress protein (hsp70) and mannosomal protein were positively correlated with heavy metal accumulation, whereas the enzyme activity showed a negative correlation with increasing heavy metal content of the slugs. PMID- 11912052 TI - Freshwater pollution biomarker: response of brain acetylcholinesterase activity in two fish species. AB - The effect of prolonged exposure at two sites along the Reconquista River (Argentina), a highly polluted peri-urban water body, on brain acetylcholinesterase (AChE, EC 3.1.1.7, acetylcholine acetylhydrolase) of two teleosts was examined. Caged Cyprinus carpio and field-captured Cnesterodon decemmaculatus were used as sentinel organisms. Eserine concentration inhibiting 50% of AChE activity (IC50) and inhibition kinetic parameters were also evaluated. Interspecies IC50 differences were found to agree with observed kinetic parameters (KA, ki and kc), indicating that carps were more sensitive to eserine. Data obtained disclosed spatial differences and demonstrated the high sensitivity of AChE activity as an exposure biomarker. Marked species-related differences were detected, showing that enzyme determination of C. decemmaculatus is more effective in highly polluted sites. Considering the river water physicochemical profile, observed changes in AChE activities can be partly attributed to long-lasting raised concentrations of dissolved heavy metals. PMID- 11912053 TI - Kinetics of metal elimination in oysters from a contaminated estuary. AB - In oysters Crassostrea gigas translocated from a metal-enriched estuary (Gironde, France) to a comparatively clean site, the Bay of Bourgneuf (France), Cd, Cu and Zn concentrations were determined monthly in the whole soft tissues, or in different fractions (cytosolic or insoluble) of gills and digestive glands. In all cases, the concentrations of all of the three metals decreased logarithmically and half-lives were always shortest for Cd (86-251 days). After 4 months, the Cd concentration had become not significantly different from the threshold considered safe for human consumption (1 mg kg(-1) wet wt.). In the digestive gland, half-lives were similar in cytosolic and insoluble fractions. In contrast, in the gills, elimination patterns differed markedly between these fractions. The long half-lives calculated for divalent metals in the insoluble fraction of the gills (1505 and 3010 days for Zn and Cu, respectively) is possibly due to a fossilization of metals in intracellular membrane-bound inclusions as shown previously in Ostrea edulis. It is interesting to underline that elimination is fastest for cytosolic metals compared to the insoluble fraction. PMID- 11912054 TI - Comparison of the biodistribution of free or liposome-entrapped Crotalus durissus terrificus (South American rattlesnake) venom in mice. AB - The local absorption rate, clearance and tissue distribution of Crotalus durissus terrificus venom, (Cdt) were examined using a two-antibody sandwich ELISA assay. We compared the biodistribution of both free or encapsulated Cdt in mice. Following subcutaneous injection of 10 microg/mouse of free Cdt (0.8 LD50), venom was detected in serum after 15 min, showed its highest level at 30 min (45+/-5 ng/ml) and was cleared from the circulation after 6 h. After 2 h of inoculation, venom was detected in the kidney (57+/-9 ng/g of tissue), spleen (18+/-4 ng/g of tissue) and brain (14+/-6 ng/g of tissue). For both subcutaneous or intravenous injection of free Cdt, venom was firstly detected in the kidney. No Cdt appeared either in the kidney, spleen, brain, or other tissues after subcutaneous inoculation of encapsulated venom even though a higher dose was used, 25 microg/mouse (2 LD50). Venom remained at the site of injection for a period of 1 week. Following intravenous injection of encapsulated venom (5 microg/mouse, 2 LD50), venom was detected in liver and spleen tissues. The biodistribution of encapsulated venom is discussed in relation to the effects of reduction of toxicity and increase of adjuvanticity. PMID- 11912055 TI - Joint action of elevated ambient nitrite and nitrate on hemolymph nitrogenous compounds and nitrogen excretion of tiger shrimp Penaeus monodon. AB - Penaeus monodon (12.13+/-1.14 g) exposed individually to six different nitrite and nitrate regimes (0.002, 0.36 and 1.46 mM nitrite combined with 0.005 and 7.32 mM nitrate), at a salinity of 25 ppt, were examined for hemolymph nitrogenous compounds and whole shrimp's nitrogen excretions after 24 h. Nitrogen excretion increased directly with ambient nitrite and nitrate. Hemolymph nitrite, nitrate, urea and uric acid levels increased, while hemolymph ammonia, oxyhemocyanin and protein were inversely related to ambient nitrite. Exposure of P. monodon to elevated nitrite in the presence of 7.32 mM nitrate did not alter hemolymph nitrite, ammonia, uric acid, oxyhemocyanin and protein levels, but caused an increase in hemolymph nitrate and a decrease in hemolymph urea as compared to exposure to elevated nitrite only. Following exposure to elevated nitrite, nitrite was oxidized to nitrate and P. monodon showed uricogenesis and uricolysis. The shrimp also used strategies to avoid joint toxicities of nitrite and metabolic ammonia by removing ammonia or reducing ammonia production under the stress of elevated nitrite. PMID- 11912056 TI - 4-Bromophenacyl bromide induces Ca2+ influx in human gingival fibroblasts. AB - 4-Bromophenacyl bromide (BPB) is generally used as a phospholipase A(2) (PLA2) inhibitor. In the present study, we demonstrate that BPB induces Ca2+ influx in human gingival fibroblasts. In fura-2-loaded human gingival fibroblasts, BPB evoked a transient increase in intracellular Ca2+ concentration ([Ca2+]i) in a dose-dependent manner. The BPB-induced Ca2+ mobilization was also shown in a single fluo-3-loaded-fibroblast. The BPB-induced increase in [Ca2+]i was completely abolished by the elimination of the external Ca2+. Ca2+ influx induced by the Ca2+-mobilizing agonist histamine was markedly enhanced in the presence of BPB. These suggest that the BPB-induced Ca2+ mobilization is due to the influx of extracellular Ca2+. However, it is unlikely that the effect of BPB is dependent on the inhibition of PLA2 activity, because other PLA2 inhibitors, such as AACOCF3, quinacrine dihydrochloride and manoalide, failed to induce Ca2+ mobilization. Chemical compounds similar to BPB, but which have no -CH2-Br at position 1 in the benzene ring failed to evoke Ca2+ mobilization, indicating that the position of -CH2--Br in BPB is important for causing the Ca2+ influx. PMID- 11912057 TI - Comparative toxinological and immunological studies on the nematocyst venoms of the Red Sea fire corals Millepora dichotoma and M. platyphylla. AB - A method appropriate for isolating of fire coral nematocysts of Millepora dichotoma (Md) and Millepora platyphylla (Mp) was described and compared with techniques that had been used before. Isolated nematocyst venoms of Md (Md-TX) and Mp (Mp-TX) were lethal to mice (had LD50 values of 0.51 and 0.21 microg/g mouse body, respectively) and displayed variable hemolytic, vasopermeable and dermonecrotic properties. The potent hemolysins of Md-TX and Mp-TX, which purified by gel filtration chromatography, possessed prominent proteins of molecular weights 35 and 31 kDa and had LD50 values 0.35 and 0.25 microg/g mouse, respectively. Hemolytic activities of crude venoms and their fraction could be inactivated using known anti-hemolytic agents. Both Md-TX and Mp-TX had distinguishable antigenic properties and their antisera raised in immunized mice and stung human were cross-reactive. ELISA assays showed an antigenic similarity among the studied fire coral homologous cytolytic counterparts. PMID- 11912058 TI - Effect of tributyltin on adenylate content and enzyme activities of teleost sperm: a biochemical approach to study the mechanisms of toxicant reduced spermatozoa motility. AB - The effects of tributyltin (TBT) on the energy metabolism and motility of fish spermatozoa were investigated in vitro in African catfish and common carp. A significant (P<0.05) decrease of the duration and the intensity of motility was observed in catfish spermatozoa exposed to 0.27 microg/l TBT for 24 h. Exposure of catfish spermatozoa to 2.7-27 microg/l TBT caused an instant decrease in ATP content. In the presence of 27 microg/l TBT approximately 55% of the initial ATP concentration in catfish semen was lost after 60 min incubation while AMP concentrations increased and the total adenine nucleotide (TAN) pool remained unchanged. The reduction in sperm ATP levels could not be attributed to cell death since viability decreased only slightly over the period of exposure. In carp by contrast, none of the adenylates concentrations studied (ATP, ADP and AMP) were affected by TBT exposure at any experimental condition. However, carp sperm motility was significantly reduced by exposure to 2.7 microg/l TBT. Among the enzymes investigated only lactate dehydrogenase (LDH) in catfish sperm was significantly (P<0.01) affected by 27 microg/l TBT treatment with a reduction in activity of approximately 75%. Compared with carp sperm before TBT exposure, that of catfish had lower adenylate contents and overall lower enzymatic activities; this explains its slower sperm velocity and shorter duration of movement as measured by computer assisted sperm analysis (CASA). The present in vitro study shows that catfish spermatozoa are more sensitive to TBT exposure (and probably to other toxicants) than those of carp. PMID- 11912059 TI - Oocyte maturation-inducing steroids in protandrous black porgy, Acanthopagrus schlegeli. AB - The study objectives aimed to investigate the maturation-inducing steroid (MIS) in marine protandrous black porgy, Acanthopagrus schlegeli. The characteristics of oocyte maturation were also described. Females were injected with two successive doses of LHRH analog (LHRH-A, 10 and 50 microg/kg of fish). The ovarian tissue was obtained at 6-h intervals for in vitro oocyte maturation. Both 17,20 beta-dihydroxy-4-pregnen-3-one (DHP) and 17,20 beta,21-trihydorxy-4-pregnen 3-one (20 beta-S) were the most effective steroids to induce in vitro maturation (e.g. germinal vesicle breakdown, GVBD) in oocytes cultured for either 24 h or 1 min. 20 beta-S had a better potency than DHP in inducing oocyte maturation. 17 hydroxyprogesterone, 11-deoxycortisol, and 20 beta-21-dihydroxy-4-pregnen-3-one also significantly induced oocyte maturation at high concentrations. The process of oocyte maturation (after the injection of LHRH analog) was founded to be divided into four stages: hormone-insensitive stage (insensitive to gonadotropin and MIS); MIS-insensitive (respond to gonadotropin, but not MIS); MIS-sensitive (respond to MIS); and spontaneous stage (GVBD in the hormone-free condition), respectively. Cycloheximide blocked GVBD at the MIS-insensitive stage, control (hormone-free), and hormone-induced GVBD at the MIS-sensitive stage in a dose dependent effect. PMID- 11912061 TI - The distribution kinetics of waterborne silver-110m in juvenile rainbow trout. AB - Juvenile rainbow trout (Oncorhynchus mykiss) were subjected to a 2-day radioactive pulse of 110mAg at 11.9 microg/l (as AgNO3), followed by a 19-day post-tracer exposure to non-radioactive Ag(I) (3.8 microg/l). The distribution of 110mAg in the gills, liver, intestine, kidney, brain and remaining carcass was investigated over a 19-day post-tracer period. Initially, the intestine contained the highest proportion of the 110mAg burden (34%), however, by day 8, less than 5% of the total radioactivity remained in this tissue. The majority of the 110mAg eliminated from the intestine appeared to distribute to the liver. Eventually, the 110mAg content in the liver accounted for as much as 65% of the total radioactivity in the fish. Apart from the liver and intestine, only the gills and carcass contained any appreciable amount (>5%) of the total body 110mAg content. Liver and gill samples were fractionated using differential centrifugation techniques to discern the subcellular distribution of 110mAg in these tissues. In the liver, the 110mAg levels in the cytosolic fraction increased from 35% to 72% of the total cellular burden between days 8 and 19, respectively. The radioactive pulse in the gills was predominantly found in a membrane compartment termed the nuclear fraction ( approximately 60% of the total). Little change was observed over time (day 8 to day 19) to the subcellular distribution of Ag in the gills. Using size-exclusion chromatography, most ( approximately 70%) of the 110mAg content in the liver cytosol eluted at a molecular weight characteristic of metallothionein. The cytosolic distribution of 110mAg in gills was quite diffuse, occurring primarily in the heavy molecular weight fractions. PMID- 11912060 TI - Species variations in the biliary and urinary excretion of arsenate, arsenite and their metabolites. AB - In most mammalian species, inorganic arsenicals are extensively biotransformed and excreted both in unchanged form and as metabolites. In the bile of rats receiving arsenate (AsV) or arsenite (AsIII) we have identified monomethylarsonous acid (MMAsIII), purportedly the most toxic metabolite of inorganic arsenic. As rats are not commonly accepted for studying arsenic metabolism, we carried out a comparative investigation on the excretion of AsV, AsIII and their metabolites in five animal species in order to determine whether they also form MMAsIII from AsV and AsIII. Anaesthetised bile duct-cannulated rats, mice, hamsters, rabbits, and guinea pigs were injected with AsV or AsIII (50 micromol/kg, i.v.) and their bile and urine was collected for 2 h. Arsenic in bile and urine was speciated by HPLC-hydride generation-atomic fluorescence spectrometry and the excretion rates of AsV, AsIII, monomethylarsonic acid (MMAsV), MMAsIII and dimethylarsinic acid (DMAsV) were quantified. All species injected with AsV excreted arsenic preferentially into urine, whereas all animals receiving AsIII, except rabbits, delivered more arsenic into bile than urine. Bile contained almost exclusively trivalent arsenic (i.e. AsIII and/or MMAsIII), whereas AsV, AsIII and DMAsV appeared in urine. Except for guinea pigs, which do not methylate arsenic, the other species formed MMAsIII and excreted it into bile. Having excreted as much as 8% of the dose of AsIII or AsV in 2 h as MMAsIII, rats were by far the most efficient producers of this supertoxic metabolite. Thus, although the rat is not a good model for studying long-term arsenic disposition, this species appears especially valuable in studies on AsIII methyltransferase and in vivo formation of MMAsIII. PMID- 11912062 TI - Seasonal short-term effects of naltrexone on LH secretion in male carp (Cyprinus carpio L.). AB - In order to evaluate the influence of the season (the stage of gonad maturity) on the modulatory role of endogenous opioid peptides in LH secretion in fish, sexually mature male carp (Cyprinus carpio L.) were intravenously injected with naltrexone-opioid receptor antagonist (5 or 50 microg kg(-1)) in the period of natural spawning (June) or gonad recrudescence (December). Moreover, the possible involvement of the dopaminergic system was studied in fish pre-treated with pimozide (dopamine receptor antagonist) and in intact fish. Blood samples were taken every minute, up to 10 min after naltrexone injection. In June, naltrexone significantly lowered LH levels in comparison to saline injected males. In December, there were no differences between saline and naltrexone-injected carps. In fish pre-treated with pimozide, neither in June nor in December were any significant differences in LH levels between control group and the groups injected with naltrexone found. The results showed that, in male carp, LH secretion under the influence of naltrexone depends on the stage of gonad maturity what suggests that the feedback of gonadal steroids on LH release could be mediated by the endogenous opioids. The role of dopamine in these processes is also discussed. PMID- 11912063 TI - Expression of HSP70 and CYP1A protein in ovary and liver of juvenile rainbow trout exposed to beta-naphthoflavone. AB - Cytochrome P4501A (CYP1A) and the 70-kDa stress protein (HSP70) were determined using Western blotting in the ovary and liver of juvenile female rainbow trout (Oncorhynchus mykiss) exposed for 4 days to beta-naphthoflavone (betaNF) following a single intraperitoneal injection. Ovarian CYP1A protein was observed in both control and betaNF-exposed fish, indicating constitutive and inducible expression of CYP1A in immature trout ovaries. CYP1A protein levels determined using densitometry were 14- and 46-fold greater in betaNF-exposed trout compared to controls in the liver and ovary, respectively. Hepatic microsomal ethoxyresorufin O-deethylase (EROD) activity, a specific catalytic marker of CYP1A, was also induced 38-fold above controls following betaNF exposure. Hepatic HSP70 protein expression was significantly higher in whole cell homogenates, but not in cytosolic fractions, collected from betaNF-exposed fish in comparison to control fish. There was no difference in ovarian HSP70 levels determined in whole cell homogenates between control and betaNF-exposed fish. The observation that unlike liver, ovarian HSP70 expression remained unchanged following induction of CYP1A protein may be related to the sensitivity of the teleost ovary to environmental toxicants that act as aryl hydrocarbon receptor agonists. PMID- 11912064 TI - The effects of ethanol on pre-synaptic components of synaptic transmission in a model glutamatergic synapse: the crayfish neuromuscular junction. AB - We have elucidated some of the mechanisms by which ethanol (EtOH) reduces synaptic efficacy at model glutamatergic synapses. The crayfish phasic and tonic neuromuscular junctions are superb models for directly assessing the effects of EtOH on pre-synaptic components of synaptic transmission. The ability to perform quantal analysis of synaptic transmission has allowed us to assess pre-synaptic alterations of release. Using this system, we report that the application of EtOH, within a range observed in intoxicated humans (44 and 88 mM), resulted in a diminution of excitatory post-synaptic potentials (EPSP) amplitudes. Additionally, using focal macro-patch recordings, quantal synaptic currents were recorded to assess the pre-synaptic component as potential target sites for EtOH's action. At the tonic neuromuscular junctions, EtOH (88 mM) reduced the probability of release (p), and in some cases, reduced the number of the release sites (n), but did not alter facilitation index nor did it affect the latency of vesicular release. At the phasic neuromuscular junction, a reduction in synaptic charge occurred during the presence of EtOH. Thus, the observed decrease in synaptic strength is at least partially attributable to a pre-synaptic alteration, specifically the release of fewer vesicles. PMID- 11912065 TI - Reported levels of alcohol consumption and binge drinking within the UK undergraduate student population over the last 25 years. AB - Results of a literature review of 18 studies investigating the drinking behaviour of undergraduate students at UK universities over a period of 25 years are presented. While comparison between studies is complicated by inconsistencies in the terms employed to describe drinking behaviour, it is concluded that significant numbers of both male and female students are reported to exceed sensible weekly consumption guidelines. Recorded levels of binge drinking among both female and male students are extremely variable between studies. Further research is needed to clarify this position. However, if the most recent research evidence is substantiated, female and male binge drinking levels may exceed those of their peers in the general population and their US counterparts. The reported ramifications of harmful drinking for the health and well-being of students are reviewed. A possible link between poor academic performance and alcohol consumption appears tenuous and merits further investigation. Evidence relevant to the view that the drinking behaviour of female students is changing is considered. PMID- 11912066 TI - Inhibitory mechanism of costunolide, a sesquiterpene lactone isolated from Laurus nobilis, on blood-ethanol elevation in rats: involvement of inhibition of gastric emptying and increase in gastric juice secretion. AB - Basic inhibitory mechanisms of costunolide and its active component, alpha methylene-gamma-butyrolactone (alpha-MGBL), on blood-ethanol elevation were investigated in rats. In normal rats, blood-ethanol elevation (30 min later) induced by 20% (v/v) ethanol [5 ml/kg, per os (p.o.)] was strongly inhibited by pretreatment (30 min earlier) with costunolide and alpha-MGBL (50 mg/kg, p.o.). In pylorus-ligated rats given ethanol, blood-ethanol level (30 min) was barely elevated compared with that of normal rats. Neither costunolide nor alpha-MGBL affected the blood-ethanol elevation in pylorus-ligated rats or that induced by intraperitoneal and intraduodenal ethanol administration. Moreover, these compounds given orally induced no irreversible changes in alcohol dehydrogenase activity in rat liver. We continuously investigated the rate of gastric emptying in rats given various test meals. Costunolide and alpha-MGBL suppressed gastric emptying in rats given 20% ethanol and 1% sodium carboxymethyl cellulose. alpha MGBL (50 mg/kg), but not costunolide, suppressed gastric emptying in 20% glucose loaded rats. In an in vitro experiment, alpha-MGBL contracted the pylorus strip at a high concentration (20 mM), which was the estimated concentration in the stomach when the substance was given orally in vivo. These findings suggested that alpha-MGBL constricted the pylorus and caused delay of gastric emptying. Moreover, both compounds increased gastric fluid secretion with pepsin and mucus. In conclusion, the inhibitory effects of costunolide and alpha-MGBL on blood ethanol elevation were based on inhibition of gastric emptying and dilution of the ethanol concentration by the increased gastric fluid. PMID- 11912068 TI - Combined effects of steroids, ethanol and protein deficiency on tissue content and urinary and faecal excretion of zinc, copper and iron. AB - This study was performed in order to determine the relative and combined effects of ethanol, a low protein diet and steroid treatment on bone, muscle, liver, and urinary and faecal excretion of zinc, copper and iron in 64 rats divided into eight groups treated following the Lieber-DeCarli liquid diet technique, with and without dexamethasone, 1 mg/l. Steroids showed a lack of effect on liver zinc, but enhanced ethanol- and low protein-mediated liver iron overload when both factors were combined. Steroids also increased muscle copper, iron and zinc, and bone copper, especially in the low protein, ethanol-fed rats. PMID- 11912069 TI - Autoshaping of ethanol drinking: an animal model of binge drinking. AB - To examine the hypothesis that Pavlovian autoshaping provides an animal learning model of drug abuse, two studies evaluated the induction of ethanol drinking by autoshaping procedures. In Experiment 1, the sipper tube conditioned stimulus (CS) contained saccharin/ethanol solution and was repeatedly paired with food as an unconditioned stimulus (US). The CS-US paired group consumed more of the 0.1% saccharin-6% ethanol solution than did the CS-US random group, revealing that autoshaping conditioned responses (CR) induce ethanol drinking not attributable to pseudo-conditioning. Experiment 2 employed saccharin-fading procedures and showed that the paired vs random group differences in ethanol drinking were maintained, even as the saccharin was eliminated from the solution. The results show that Pavlovian autoshaping procedures induce high volumes of ethanol drinking when the presentation of a sipper tube containing an ethanol solution precedes the response-independent delivery of food. The high volume of ethanol consumed in a brief period of time suggests that Pavlovian autoshaping may be a model of binge drinking. PMID- 11912067 TI - Gamma-hydroxybutyric acid versus alcohol preference in Sardinian alcohol preferring rats. AB - Previous experiments demonstrated that the selectively bred Sardinian alcohol preferring (sP) rats possess a genetically based proclivity to consume pharmacologically relevant doses of gamma-hydroxybutyric acid (GHB). The present study was aimed at comparing the reinforcing properties of GHB and ethanol, measuring the propensity of sP rats to consume GHB and ethanol when both drugs were concomitantly available. Initially, two groups of sP rats (ethanol-naive and ethanol-experienced, respectively) were forced to consume GHB in order to help them discover the reinforcing properties, which could then prevail over the unpleasant taste of the GHB solution. Subsequently GHB (at concentrations increasing from 1 to 6% w/v) was offered in free choice with water and all rats consumed pharmacologically relevant amounts of GHB. Finally, under the free choice regimen between GHB (presented to each rat at its preferred concentration), ethanol and water, daily ethanol intake averaged approximately 6 g/kg (i.e. the amount of ethanol usually consumed by sP rats), whereas GHB intake declined by approximately 75%. In the few rats showing a high intake of GHB, ethanol intake was not altered. No difference in GHB drinking behaviour was ever recorded between ethanol-naive and ethanol-experienced rats. The results of the present study demonstrate that freely available GHB is not capable of altering ethanol preference and consumption in sP rats and suggest that the postulated reciprocal substitutability of the two drugs does not completely include the reinforcing properties, at least in sP rats and when oral self-administration of GHB is considered. The results also provide a model of the low abuse liability of GHB observed in human alcoholics. PMID- 11912070 TI - Novelty seeking and harm avoidance in relation to alcohol drinking in intact rats and following axon-sparing lesions to the amygdala and ventral striatum. AB - Our previous research has pointed to a behavioural pattern in rats similar to the one seen in the prototypic Type 2 alcoholic, where male rats bearing basal forebrain excitotoxin axon-sparing lesions show an increased alcohol intake, lowered responsiveness to threat (harm avoidance) and an enhanced tendency to exploring novel surroundings (novelty seeking). The purpose of the present study was to: (1) examine whether excessive alcohol-consuming rats have a diminished response to potential danger and an enhanced tendency to explore novel surroundings; (2) investigate a possible relationship between the above-mentioned behavioural parameters and two of the emotional motor systems in the basal forebrain, i.e. the extended amygdala and the ventral striato-pallidal system. Thus, we compared the effect of ibotenic axon-sparing lesions (3.5 microg/0.35 microl) to the temporal part of the amygdala with lesions in the ventral striatum on alcohol intake, harm avoidance (stretched attend postures) and novelty seeking (nose pokes), before and after surgery. Our results suggest that: (1) intact animals high in novelty seeking drink more alcohol than rats low on this trait; (2) lesions of the amygdala decrease harm avoidance and fluid intake, especially in animals showing above-average levels of risk assessment pre-operatively; (3) neuronal loss in the ventral striatum stimulates novelty seeking and alcohol intake, particularly in rats which initially show less novelty seeking than average. PMID- 11912071 TI - Alcohol consumption during murine acquired immunodeficiency syndrome accentuates heart pathology due to coxsackievirus. AB - Alcohol, especially after prolonged and excessive consumption, results in marked alteration of host immunity and increased susceptibility to infection. To determine whether ethanol consumption exacerbates coxsackievirus B3 cardiomyopathy during murine acquired immunodeficiency syndrome (AIDS), female C57BL/6 mice were infected with LP-BM5 retrovirus and administered 40% ethanol both in water and in solid agar-based form. Cardiac histopathology was semi quantitatively assessed for lesion severity and induced production of splenocyte: interleukin (IL)-2, IL-4, IL-6, tumour necrosis factor-alpha and interferon-gamma were determined. Ethanol consumption during murine retrovirus infection increased coxsackievirus-induced myocarditis in 85% of the animals and also exacerbated the lesion severity. Mice infected with retrovirus and co-infected with coxsackievirus showed significant heart lesions. Retrovirus infection suppressed Th1 responses, causing cytokine dysregulation and immunosuppression, which facilitated coxsackievirus-induced myocarditis. Our data suggest that ethanol consumption heightens the cytokine imbalance to favour a Th2 response by enhancing Th2 and/or by suppressing Th1 function. In conclusion, murine AIDS facilitated severe cardiotoxicity during coxsackievirus infection, while non retrovirus-infected mice were resistant. These effects were accentuated by ethanol consumption. PMID- 11912073 TI - Effects of acute alcohol intoxication on pituitary-gonadal axis hormones, pituitary-adrenal axis hormones, beta-endorphin and prolactin in human adults of both sexes. AB - - The effects of acute alcohol intoxication (AAI) on the pituitary-gonadal axis hormones, and the possible contribution of pituitary-adrenal axis hormones, beta endorphin and prolactin to alcohol-induced dysfunction of pituitary-gonadal axis hormones were studied in adult men and women. Blood samples were drawn from adults of both sexes who arrived at the emergency department with evident behavioural symptoms of drunkenness (AAI) or from adult volunteers with nil consumption of alcohol (controls). Our results demonstrated that AAI produces a high increase in plasma prolactin, corticotropin (adrenocorticotropic hormone, ACTH), and cortisol in adults of both sexes, a decrease in luteinizing hormone levels only in men, an increase in dehydroepiandrosterone-sulphate (DHEAS) and a contradictory behaviour of testosterone according to gender, with increased plasma testosterone in women and a decrease in men. ACTH and prolactin correlated positively with cortisol, DHEAS and testosterone in women, which suggests that prolactin and ACTH could contribute to stimulated adrenal androgen production. In contrast, the decrease in testosterone and increase in beta-endorphin in men suggests that AAI could have an inhibitory effect on testicular testosterone, perhaps mediated by beta-endorphin. Our results suggest that the effect of alcohol on pituitary-gonadal axis hormones in humans could depend on the gender and degree of sexual maturity of the individual. PMID- 11912072 TI - Short-term methadone administration reduces alcohol consumption in non-alcoholic heroin addicts. AB - - Methadone, a synthetic opioid agonist, is commonly used for the treatment of heroin dependence. Depending on how alcohol addiction is defined, rates of alcoholism vary among those attending methadone maintenance treatment (MMT) programmes. Most of the current literature has shown that alcohol consumption increases during medium- or long-term MMT. However, up to now, no data have been reported on changes in alcohol intake among a population of heroin addicts with no alcohol-dependence diagnosis after short-term methadone administration. Thus, the aim of our study was to investigate alcohol consumption changes in a population of non-alcoholic heroin addicts during the first 4 weeks of a treatment programme (TP). The TP consisted of either MMT or non-methadone maintenance treatment (N-MMT) with a minimum duration of 1 year. A total of 359 heroin-addicted out-patients [274 males (76.3%)], all of whom met DSM-IV criteria, were enrolled in the study, over a period of 4 months. Out of these 359 patients, 32 subjects (8.9%) dropped out, whereas 327 subjects (91.1%; 249 males) continued the TP [105 (32.1%; 78 males) in the MMT group and 222 (67.9%; 171 males) in the N-MMT group]. A significant reduction in daily alcohol intake was observed in the MMT group, but not in the N-MMT group after the first 4 weeks of the TP. The results of the present study suggest a possible effect of short-term methadone administration in reducing alcohol consumption in a population of non alcoholic heroin-addicted patients. PMID- 11912074 TI - No association between metabotropic glutamate receptors 7 and 8 (mGlur7 and mGlur8) gene polymorphisms and withdrawal seizures and delirium tremens in alcohol-dependent individuals. AB - - Up-regulation of the glutamatergic neurotransmission from chronic ethanol intoxication may cause a hyperexcitable state during alcohol withdrawal that may lead to seizures and delirium tremens. The aim of our study was to evaluate the association between a history of alcohol withdrawal-induced seizures and delirium tremens and a mGlurR7 (Tyr433Phe); and a mGlurR8 (C2756T) metabotropic glutamate receptor polymorphism in alcoholics compared to controls. A total of 182 patients meeting DSM-IV alcohol dependence criteria and 117 controls, both groups being of German descent, were investigated. mGluR7 and mGluR8 polymorphisms were determined using polymerase chain reaction of lymphocyte DNA. History of alcohol withdrawal-induced delirium tremens and seizures were obtained using the Semi Structured Assessment of Genetics in Alcoholism (SSAGA). Data were cross-checked with inpatients' clinical files. No significant associations were obtained between both receptor polymorphisms and alcohol withdrawal-induced seizures and delirium tremens. The negative results in this study question the role of these polymorphisms in the pathogenesis of alcohol withdrawal-induced seizures and delirium tremens. PMID- 11912075 TI - In vitro effects of ethanol, acetaldehyde and fatty acid ethyl esters on human erythrocytes. AB - In vitro experiments were performed to determine if ethanol was metabolized by human erythrocytes and to investigate if ethanol or its metabolites, acetaldehyde and fatty acid ethyl esters, affected erythrocyte morphology and stability. No detectable metabolism of ethanol was found in erythrocytes, although ethanol itself caused an elevated rate of spontaneous haemolysis in erythrocyte preparations. Physiologically attainable levels of ethanol were found to stabilize erythrocytes against haemolysis induced by sodium hypochlorite, and the presence of ethanol caused a decrease in erythrocyte reactive oxygen species levels, although the mechanism for such a process is unknown. Both physiologically attainable and higher levels of acetaldehyde had no effects on erythrocyte morphology and stability even after a 16 h exposure. Fatty acid ethyl esters caused structural changes and instability in erythrocytes in vitro, but whether such changes occur in vivo has not been established. The results of these studies suggest that the deleterious effects of ethanol consumption on erythrocytes in vivo may be, at least in part, the result of direct effects of unmetabolized ethanol on erythrocyte components. PMID- 11912077 TI - Religious beliefs and practice, and alcohol use in Thai men. AB - Buddhism, the Thai state religion, teaches that use of intoxicants should be avoided. Nonetheless, many Thai people drink alcohol, and a proportion are alcohol-dependent or hazardous or harmful drinkers. This study examines the relationship between Buddhist upbringing and beliefs and alcohol use disorders in Thai men. Three groups, comprising 144 non/infrequent/light drinkers, 77 hazardous/ harmful drinkers and 91 alcohol dependents were interviewed regarding their early religious life and current religious practices and beliefs. No protective association was shown between early religious life and later alcohol use disorders; indeed, having lived as a boy in a temple for a period was commoner in those with adult alcohol problems. Few subjects reported frequent involvement in current religious activities (9, 8 and 6% in the non/infrequent/light drinkers, hazardous/harmful drinkers, and alcohol dependents respectively). Hazardous/harmful drinkers [odds ratio (OR) = 0.4, 95% confidence interval (CI) = 0.2-0.9] and alcohol dependents (OR = 0.5, 95% CI = 0.2-0.9) were less likely to report being moderately to strongly religious, than were non/infrequent/light drinkers. Understanding the association between religious beliefs and drinking behaviour can potentially assist in the development of prevention and treatment programmes. PMID- 11912076 TI - Patterns of alcohol drinking in a population of young social drinkers: a comparison of questionnaire and diary measures. AB - Studies of alcohol use often depend on self-reported alcohol intake measured by quantity/frequency questionnaires. Previous research has shown that alcohol consumption may be underestimated by this type of retrospective questionnaire. The primary aim of this study was to compare the accuracy of an Alcohol Use Questionnaire (AUQ) with a 4-week diary account. A further aim was to explore patterns of drinking in young social drinkers, with particular attention to binge drinking, which has been suggested as a factor in increasing the risk of alcohol dependency. University students completed the AUQ in the laboratory. They were then asked to keep a record of their alcohol, nicotine and caffeine consumption over a 4-week period (diary). The questionnaire and the diaries were compared on factors of alcohol intake (units per week) and patterns of drinking behaviour (speed of drinking, number of times being drunk and percentage of times getting drunk when drinking). The two measures (AUQ and diary) were highly correlated on alcohol consumption and the other questions relating to drinking behaviour. However, differences were found between the two measures on alcohol intake, speed of drinking (drinks per hour) and number of times being drunk. Alcohol consumption was underestimated by approximately 12% on the questionnaire, and, when the accuracy of estimation of drinking habits was examined, it was found that high drinkers tended to underestimate their drinking behaviour, whereas lower drinkers tended to overestimate. The results suggest that the AUQ can be used with a reasonable degree of confidence, bearing in mind the tendency for high drinkers to underestimate consumption and drinking behaviour. Relationships between 'binge scores', beverage specificity and alcohol consumption support the idea that the criteria for binge drinkers should be based on patterns of drinking rather than alcohol consumption. PMID- 11912078 TI - Prevalence of problem drinking in a Venezuelan Native American population. AB - This is the first study of alcohol-related problems among a Latin American indigenous population using the Alcohol Use Disorders Identification Test (AUDIT). A randomly selected community sample consisting of 3% of the adult population of the tribe completed oral interviews (n = 105 adults, completion rate 86%). The majority of both men (98%) and women (53%) had drunk alcohol at some time in their lives, with 94 and 26% respectively having consumed alcohol within the past 12 months. Using a cut-off score of 8 for the AUDIT, 86.5% of all men and 7.5% of all women were found to be problem drinkers. Focus group discussions revealed that traditional patterns of binge drinking of corn liquor had gradually been replaced by consumption of commercial beer and rum at more frequent intervals and with more negative social consequences. This male population demonstrates one of the highest prevalence rates for problem drinking reported in the world literature. Both the magnitude of problems detected and participants' concerns about them suggest that broad-scale interventions are warranted at the community level. PMID- 11912079 TI - Genes, logic and statistics: response to a review. PMID- 11912081 TI - No evidence of impaired neurocognitive performance in collegiate soccer players. AB - A high incidence of cerebral concussion has been reported among soccer players. We studied whether long-term or chronic neuropsychological dysfunction was present in collegiate soccer players. Two hundred forty subjects from a National Collegiate Athletic Association division I institution were stratified into three groups: soccer athletes (91), nonsoccer athletes (96 women's field hockey, women's lacrosse, and baseball players), and controls (53 college students). Subjects completed a concussion history questionnaire and underwent preseason baseline neuropsychological testing before the start of either the freshman or sophomore year. Data were collected on the results of six neuropsychological tests and from a concussion history questionnaire for number of previous concussions, Scholastic Aptitude Test results, and exposure to soccer and heading. Despite an average of 15.3 seasons of soccer exposure and a higher prevalence of previous concussions, the soccer athletes did not demonstrate impaired neurocognitive function or scholastic aptitude when compared with the nonsoccer athletes or the student nonathletes. Additionally, there was no significant relationship between a history of soccer-related concussion and either neurocognitive performance or scholastic aptitude. Neither participation in soccer nor a history of soccer-related concussions was associated with impaired performance of neurocognitive function in high-level United States soccer players. PMID- 11912082 TI - Tennis after total knee arthroplasty. AB - In this study, patients who played tennis after undergoing a total knee arthroplasty were analyzed in terms of their functional abilities and degree of satisfaction. The patients were recruited by means of a questionnaire that was sent to players from lists supplied by the United States Tennis Association. The study group consisted of 28 men and 5 women (46 total knee replacements) with a mean age of 64 years. Only 21% (7 of 33) of the patients' surgeons approved of their patients undertaking tennis activity, with 45% (15 of 33) recommending only doubles tennis. At both 1 year and a mean of 7 years after arthroplasty, players were playing both singles and doubles tennis approximately three times per week (range, one to seven). All tennis players polled were satisfied with their knee arthroplasties and their ability to resume playing tennis. Because the study patients played at a high level, future studies are needed to determine the effect of tennis on the general population, which does not play at such a uniformly high level. The long-term (15 to 20 years) effect of tennis activity on the clinical and radiologic outcome of total knee arthroplasty also needs to be determined. PMID- 11912083 TI - Compaction versus extraction drilling for fixation of the hamstring tendon graft in anterior cruciate ligament reconstruction. AB - Initial strength of quadrupled hamstring tendon grafts fixed with titanium interference screws was assessed in 30 pairs of porcine tibiae. Bone tunnels were drilled with either compaction drilling (stepped routers) or conventional extraction drilling (cannulated drill bits). Fifteen pairs of specimens were subjected to a single-cycle load-to-failure test, while the rest underwent a cyclic-loading test to further assess the quality of the fixation. No significant difference between the two drilling techniques was found with regard to yield load, displacement at yield load, stiffness, or mode of failure. Porcine trabecular bone mineral density was determined using peripheral quantitative computed tomography and compared with that of young women and men at a site corresponding to that of the tibial bone drill hole of an anterior cruciate ligament reconstruction. There was a significant difference between the two species (210 +/- 45 mg/cm(3) in porcine tibial bone versus 129 +/- 30 mg/cm(3) in women and 134 +/- 34 mg/cm(3) in men), suggesting that porcine knee specimens may have limitations in studies of graft fixation in anterior cruciate ligament reconstruction. We found no difference between extraction and compaction drilling in initial fixation strength of a hamstring tendon graft for anterior cruciate ligament reconstruction using a porcine model. PMID- 11912084 TI - Long-term results of meniscal allograft transplantation. AB - Between May 1984 and December 1986, 23 patients with a history of medial meniscectomy and anterior knee instability were entered into a long-term prospective study of the results of medial meniscal transplantation combined with reconstruction of the anterior cruciate ligament. In 17 cases a lyophilized meniscal allograft was used and in 6 cases a deep-frozen meniscal allograft was used. The patients' clinical outcomes were evaluated 3 and 14 years postoperatively by clinical assessment, Lysholm score, radiographs, magnetic resonance imaging, arthrography, and, in some cases, arthroscopy. Two anterior cruciate ligament reconstruction control groups were used for comparison, one group having previously undergone meniscectomy and one with intact menisci. The follow-up rate was 100% after 14 years. The Lysholm score was 84 +/- 12 points at 3 years postoperatively and 75 +/- 23 points at 14 years. Patients with deep frozen meniscal transplants generally had better results than patients with lyophilized meniscal transplants. Magnetic resonance imaging evaluation showed good preservation of the deep-frozen meniscal transplants, even after 14 years. The lyophilized meniscal transplants were reduced in size at the second-look arthroscopy and as seen on magnetic resonance imaging examination. When the control groups were compared with the study group, the deep-frozen meniscal allografts were found to be more comparable with an intact meniscus and the lyophilized meniscal allografts were more comparable with the control group knees that had undergone meniscectomy. PMID- 11912085 TI - The effect of the menstrual cycle on anterior cruciate ligament injuries in women as determined by hormone levels. AB - Anterior cruciate ligament injury rates are reported to be two to eight times higher in women than in men within the same sport. Because the menstrual cycle with its monthly hormonal fluctuations is one of the most basic differences between men and women, we investigated the association between the distribution of confirmed anterior cruciate ligament tears and menstrual cycle phase. Sixty nine female athletes who sustained an acute anterior cruciate ligament injury were studied within 24 hours of injury at four centers. The mechanism of injury, menstrual cycle details, use of oral contraceptives, and history of previous injury were recorded. Urine samples were collected to validate menstrual cycle phase by measurement of estrogen, progesterone, and luteinizing hormone metabolites and creatinine levels at the time of the anterior cruciate ligament tear. Results from the hormone assays indicate that the women had a significantly greater than expected percentage of anterior cruciate ligament injuries during midcycle (ovulatory phase) and a less than expected percentage of those injuries during the luteal phase of the menstrual cycle. Oral contraceptive use diminished the significant association between anterior cruciate ligament tear distribution and the ovulatory phase. PMID- 11912086 TI - The clinical significance of anterior horn meniscal tears diagnosed on magnetic resonance images. AB - We assessed the accuracy of magnetic resonance imaging in detecting clinically significant lesions of the anterior horn of the meniscus by reviewing 947 consecutive knee magnetic resonance imaging reports. Of these, 76 (8%) indicated a tear of the anterior horn of the medial or lateral meniscus. Thirty-one of these 76 patients underwent a subsequent arthroscopic examination, and their operative reports were reviewed. The 45 patients who were not examined arthroscopically were contacted and interviewed for clinical follow-up. Among the 31 patients who underwent arthroscopic examination, 8 anterior horn tears were noted in the predicted area (26% true-positive results), 23 patients had intact anterior horns (74% false-positive results), and 18 had normal intact menisci in all zones. Of the 45 patients who did not undergo arthroscopic surgery, 6 had isolated anterior horn tears reported on magnetic resonance imaging, and 5 of the 6 were asymptomatic at follow-up. The other 39 patients had multiple pathologic conditions noted on the magnetic resonance imaging report and continued to report knee pain at the follow-up interview. Increased signal intensity at the anterior horn of the meniscus seen on magnetic resonance imaging commonly does not represent a clinically significant lesion. We recommend correlation with the physical examination when interpreting this "positive" finding on knee magnetic resonance imaging examinations. PMID- 11912087 TI - Epidemiologic analysis of injury in five years of Canadian professional rodeo. AB - Longitudinal studies of rodeo injuries are rare. We prospectively investigated injuries in professional rodeo in Canada over a 5-year period. Our specific interests included injury incidence density in specific rodeo events, risk factors such as past injury, and the incidence of head injury. Of 323 professional rodeos from 1995 through 1999, 63 rodeos provided a convenience sample. These rodeos were selected because the Canadian Professional Rodeo Sport Medicine Team was in attendance at these events, thus providing both competitor health care and data collection. Four hundred fifty-one injuries were reported during 30,564 competitor-exposures. The greatest injury frequency and injury incidence density were in the rough stock events (bull riding, bareback riding, and saddle bronc). Bull riding accounted for the greatest injury frequency (141) and incidence density (32.2 injuries per 1000 competitor-exposures). Bull riding had a relative injury risk of 1.32 when compared with bareback riding; bareback riding had a relative injury risk of 1.39 when compared with saddle bronc riding. Concussions accounted for 8.6% of all reported injuries. Concussions and other head injuries (65) were second only to knee injuries (76) in frequency of injury to specific body parts. This concussion frequency is higher than has previously been reported. PMID- 11912088 TI - Hamstring muscle strain recurrence and strength performance disorders. AB - We determined the frequency of strength disorders in 26 athletes with a history of hamstring muscle injury and recurrent strains and discomfort. We also assessed the effectiveness of rehabilitation to correct muscle performance. After concentric and eccentric isokinetic assessment, 18 athletes were found to have strength deficits, as determined by statistically selected cutoffs of peak torque, bilateral differences, and the flexors/quadriceps ratio. The discriminating character of the eccentric trial was demonstrated, combining a preferential eccentric peak torque deficit and a significant reduction of the mixed eccentric flexors/concentric quadriceps ratio. The athletes with muscle imbalances followed a rehabilitation program individually adapted from their strength profile. Treatment length was from 10 to 30 sessions and resulted in isokinetic parameter normalization in 17 of 18 subjects. Isokinetically corrected subjects were observed for 12 months after return to athletics. None sustained a clinically diagnosed hamstring muscle reinjury. Subjective intensity of pain and discomfort were significantly reduced, and they all returned to their prior level of competition. These results demonstrate that persistent muscle strength abnormalities may give rise to recurrent hamstring injuries and discomfort. An individualized rehabilitation program emphasizing eccentric training based on specific deficits contributes to a decrease in symptoms on return to sports. PMID- 11912089 TI - Regeneration of the semitendinosus tendon after its use in anterior cruciate ligament reconstruction: a histologic study of three cases. AB - We describe the histologic course of regenerated tissue simulating the semitendinosus tendon in three patients after anterior cruciate ligament reconstruction with doubled semitendinosus and gracilis tendons. Specimens were retrieved during revision surgery for removal of tibial hardware at 6, 24, and 27 months, respectively. Resisted flexion during physical examination in all patients revealed a "string" on the posteromedial aspect of the knee that was quite evident, suggesting the presence of a regenerated semitendinosus tendon. At surgery, a well-defined fibrous band was identified about 3 cm proximal and medial to the pes anserinus, reproducing the pathway of the native semitendinosus tendon. In the specimen retrieved 6 months postoperatively, a prominent fibroblastic proliferation was observed together with a few vessels surrounded by fibrous tissue; only a few bundles of well-oriented collagen fibers were present, together with scattered rows of spindle-shaped cells. In both of the specimens retrieved 2 years postoperatively, all of the central, thicker portion of each specimen was occupied by well-oriented tendon-like fibers together with uniformly distributed spindle-shaped cells that had the appearance of mature tenocytes arranged in orderly rows. These findings help us to understand the progression of the healing process at the harvest site and the nature of the regenerated semitendinosus tendon, which has previously been investigated only by studies of clinical and functional outcomes and by imaging studies. PMID- 11912090 TI - Anterior cruciate ligament reconstruction with a reharvested bone-patellar tendon bone graft. AB - We sought to determine the results in patients who underwent revision anterior cruciate ligament reconstruction with a reharvested bone-patellar tendon-bone graft. Preoperative magnetic resonance imaging of the patellar tendon was used to determine the width of the tendon and the quality of the tissue. The section of the tendon harvested included 7 to 8 mm of the regenerated tendon and 2 to 3 mm of previously untouched tendon either medial or lateral to the healed tissue. The mean time from primary graft harvest to reharvest was 71.0 +/- 44 months (range, 32 to 180). An accelerated rehabilitation program was used postoperatively. Objective follow-up of more than 2 years after surgery was available for 8 patients at a mean of 49.0 +/- 15.4 months. The average range of motion in the anterior cruciate ligament-reconstructed knee was 6/0/133 at 1 month postoperatively and 6/0/145 at long-term follow-up. The mean manual maximum KT 1000 arthrometer difference between knees was 1.6 +/- 1.2 mm. The mean isokinetic quadriceps muscle strength of the reharvested graft donor knee was 103% +/- 7% of that of the opposite knee. Subjective scores were obtained from 11 patients (12 knees) at a mean of 51.5 +/- 35.7 months (range, 24 to 150) after surgery; the mean score was 89.3 +/- 6.9 points. We conclude that with appropriate preoperative planning, a reharvested patellar tendon can be used for revision anterior cruciate ligament reconstruction to obtain reliable stability and strength postoperatively. PMID- 11912091 TI - A prospective randomized comparison of patellar tendon versus semitendinosus and gracilis tendon autografts for anterior cruciate ligament reconstruction. AB - Seventy patients with patellar tendon or hamstring tendon autografts for single incision anterior cruciate ligament reconstruction were evaluated at least 2 years after surgery. All reconstructions were performed by the same surgeon, and metal interference screws were used for fixation of all grafts. No significant differences were noted between groups for Lysholm score, reduction in activity, KT-1000 arthrometer findings, quadriceps muscle size, return to sports, or ability to jump and do hard cuts and pivots. Significantly more patients in the patellar tendon group had patellofemoral pain at 6 months after surgery than did the hamstring tendon patients (48% versus 20%), and at last follow-up the incidence of patellofemoral pain was 42% and 20%, respectively. Fourteen patients in the patellar tendon group and seven in the hamstring tendon group had loss of motion (approximately 5 degrees ). Four patients (two in each group) had treatment failures and their results were not included in the clinical examination data. At 2 years' follow-up, 97% of patients with patellar tendon grafts and 100% of patients with hamstring tendon grafts rated their results as good or excellent. We found that hamstring tendon grafts performed similarly to patellar tendon grafts, although fewer patients in the hamstring tendon group had patellofemoral pain and loss of motion. PMID- 11912092 TI - Radiofrequency electrothermal shrinkage of the anterior cruciate ligament. AB - The efficacy of electrothermal collagen shrinkage in the treatment of patients with anterior cruciate ligament laxity was evaluated. Eighteen patients who had continuity of the anterior cruciate ligament but had symptomatic laxity were treated with arthroscopic electrothermal shrinkage of the anterior cruciate ligament using a monopolar radiofrequency probe. The mean length of follow-up in patients whose stability was maintained was 20.5 months. Seven of the patients had undergone previous reconstruction, four with patellar tendon graft and three with quadrupled hamstring tendon graft. Laxity was chronic in nine patients and acute in nine. The KT-1000 arthrometer results at 1 month postoperatively revealed decreased anterior excursion, with an average side-to-side difference of 1.9 mm. However, 11 patients had a failed result at an average 4.0 months. Of the seven patients with successful results, six had native ligaments and had been treated for acute laxity and one had a patellar tendon graft and had been treated for chronic laxity. Even with the short-term follow-up in our study, it is evident that thermal shrinkage using radiofrequency technology has limited application for patients with anterior cruciate ligament laxity. Although it may be useful in treating patients with an acutely injured native anterior cruciate ligament, further study is needed to see if the ligament stretches out over time or is at increased risk of reinjury. PMID- 11912093 TI - Results of arthroscopic treatment of posterosuperior glenoid impingement in tennis players. AB - Twenty-eight tennis players with symptomatic posterosuperior glenoid impingement limiting their participation underwent arthroscopic debridement of the supraspinatus tendon and glenoid lesions associated with this diagnosis after nonoperative treatment had failed. The dominant extremity was affected in all patients; the patients' average age was 26.9 years. Eighteen patients participated at the highest level of competition for their age, and the remaining patients participated at the intermediate level. Patients were evaluated at an average of 45.7 months after surgery by physical examination, an activities questionnaire, a subjective result questionnaire, and a questionnaire regarding their return to activity. Postoperatively, the patients averaged 26.9 of 30 possible points on the activities questionnaire. Twenty-three of the patients were subjectively satisfied with the surgical result. Twenty-two patients had returned to tennis. Despite their return, 20 of the 22 patients reported some persistent pain with participation. To our knowledge, this is the first report detailing the results of operative treatment for posterosuperior glenoid impingement in a population limited to tennis players. Even though the results are encouraging in terms of the high number of patients returning to tennis, the effects of this persistent pain with activity, although diminished in severity, on long-term participation is unknown. PMID- 11912094 TI - The effect of injury to the posterolateral structures of the knee on force in a posterior cruciate ligament graft: a biomechanical study. AB - To determine whether untreated grade 3 posterolateral knee injuries contribute to a significant increase in force on a posterior cruciate ligament reconstruction graft, we measured the force on the graft during joint loading of a posterior cruciate ligament-reconstructed knee with otherwise intact structures and then selectively cut the popliteofibular ligament, popliteus tendon, and the fibular collateral ligament. A posterior cruciate ligament reconstruction was performed in eight fresh-frozen cadaveric knees. One end of the graft was fixed to a tensioning jig with a load cell used to measure force in the graft as loads were applied to the knee. The force on the graft was significantly higher with the posterolateral structures cut during varus loading at 30 degrees, 60 degrees, and 90 degrees of flexion than it was in the same joint under the same loading conditions but with the posterolateral structures intact. Additionally, coupled loading of posterior drawer force and external tibial torque at 30 degrees, 60 degrees, and 90 degrees significantly increased force on the graft with the posterolateral structures cut. There was no significant increase in force on the graft under any condition with a posterior force, valgus force, or internal and external tibial torque applied alone. A significant increase in force occurs in a posterior cruciate ligament graft in knees with deficient posterolateral knee structures. We recommend that in knees with grade 3 posterolateral injuries and evidence of varus or coupled posterior-external rotation instability the posterolateral structures be repaired or reconstructed at the time of posterior cruciate ligament reconstruction to decrease the chance of later graft failure. PMID- 11912095 TI - Fatigue testing of suture anchors. AB - In a porcine tibia model, we subjected widely used anchor-suture combinations to a fatigue-testing protocol. The Ethibond No. 2 suture was the weakest part of the anchor-suture combinations when they were loaded to failure by a single pull. Under cyclic-loading conditions, fixation strength was decreased compared with single-pull tests. The suture/anchor interface was identified as the weakest link in the Mitek GII/No. 2 combination and in the Zimmer Statak 3.5/No. 2 combination. In most cases the suture was worn through at the eyelet. Threading the GII anchor with a No. 5 suture and use of larger anchors in combination with No. 2 sutures increased the fatigue strength. Suture breakage at the knot was the predominant failure mode for biodegradable anchors inserted into cortical bone. The highest fatigue strength was seen for the Super Anchor/No. 5 combination when the anchor was inserted in cortical bone. Fatigue testing is crucial for evaluation of suture anchors and should be performed along with single-pull testing. The mechanical performance of a suture anchor threaded with a defined suture depends on several key factors: the pullout strength of the anchor, the tensile strength of the suture, and the interaction of anchor and suture at the eyelet (suture/anchor interface). PMID- 11912096 TI - Injury incidence and prevalence among elite weight and power lifters. AB - The purpose of this study was to investigate the incidence and prevalence of injuries among elite weight lifters and power lifters, with a special focus on shoulder injuries and possible injury-provoking exercises. In 1995, a questionnaire was administered to 110 male and female elite lifters to evaluate injuries and training characteristics. A follow-up of the athletes from 1995 was conducted in 2000, and a new 2000 elite group was also queried. In 1995 and again in 2000, the athletes sustained, on average, 2.6 injuries per 1000 hours of activity. Most common in 1995 were low back injuries, with an injury rate of 0.43 per 1000 hours, and shoulder injuries, with a rate of 0.42 per 1000 hours. Shoulder injuries dominated in 2000, with an injury rate of 0.51 per 1000 hours of activity. There was a difference in injury pattern between weight lifters, who mostly sustained low back and knee injuries, and power lifters, in whom shoulder injuries were most common. No correlation was found between shoulder injuries and any specific exercise. Although the total injury rate was the same during the two periods of study, the rate of shoulder injuries had increased. PMID- 11912097 TI - The partial-thickness rotator cuff tear: is acromioplasty without repair sufficient? AB - We evaluated the clinical outcome of arthroscopic acromioplasty and debridement in 162 patients who had either normal rotator cuffs, grade 1 (frayed tendon) partial-thickness tears, or grade 2 (less than 50% of the tendon) partial thickness tears. The mean time from surgery to the response to the L'Insalata outcome questionnaire was 52.7 months (4.5 years) among the 105 respondents (107 shoulders). The mean score was 90 points; eight patients (8%) scored less than 70 points (range, 30 to 65.5), and their treatment failed early on. The patients with grade 2B (bursal) partial-thickness rotator cuff tears had a significantly higher failure rate (38%). Although the clinical outcome of patients with partial thickness tears of the rotator cuff comprising less than 50% of the tendon (grade 1 and 2) was not significantly different from that of patients without partial rotator cuff tears, the subgroup of patients with grade 2B partial tears had a statistically significantly higher failure rate and may have been better served with primary repair. With follow-up to 10 years, there was no evidence that clinically relevant or symptomatic intrinsic rotator cuff pathologic conditions progress in those patients with partial-thickness tears treated with arthroscopic anterior acromioplasty. PMID- 11912098 TI - A comparison of knee kinetics between male and female recreational athletes in stop-jump tasks. AB - We compared the knee kinetics of 10 male and 10 female recreational athletes (aged 19 to 25 years) performing forward, vertical, and backward stop-jump tasks. Three-dimensional videography and force plate data were used to record the subjects' performance of the three stop-jump tasks, and an inverse dynamic procedure was used to estimate the knee joint resultant forces and moments. Women exhibited greater proximal anterior shear force than did men during the landing phase. All subjects exhibited greater proximal tibia anterior shear force during the landing phase of the backward stop-jump task than during the other two stop jump tasks. Women also exhibited greater knee extension and valgus moments than did men during the landing phase of each stop-jump task. Men exhibited greater proximal tibia anterior shear force than did women during the takeoff phase of vertical and backward stop-jump tasks. These results indicate that female recreational athletes may have altered motor control strategies that result in knee positions in which anterior cruciate ligament injuries may occur. The landing phase was more stressful for the anterior cruciate ligament of both women and men than the takeoff phase in all stop-jump tasks. Technical training for female athletes may need to be focused on reducing the peak proximal tibia anterior shear force in stop-jump tasks. Further studies are needed to determine the factors associated with the increased peak proximal tibia anterior shear force in female recreational athletes. PMID- 11912099 TI - Analysis of pre- and postexercise compartment pressures in the medial compartment of the foot. AB - Growing awareness of chronic exertional compartment syndrome of the foot has led to the need for site-specific analysis of normative resting and postexertional intracompartment pressures. Thirty-four asymptomatic athletes underwent pressure testing of the medial foot compartment with an intracompartmental pressure measurement apparatus. Pressure measurements were recorded before exercise and after 20 minutes of running. Mean resting intracompartment pressure was 7.7 mm Hg, whereas mean pressures 1 and 5 minutes after exercise were 19.1 mm Hg and 10.7 mm Hg, respectively. These data were then compared with data from compartment pressure studies performed in nine symptomatic subjects, each with a clinical history suggestive of chronic exertional compartment syndrome of the medial foot compartment. The results demonstrated normative compartment pressures of the medial foot compartment are comparable with previously measured pressures of the leg. This study shows that previously defined criteria for diagnosis of chronic exertional compartment syndrome of the leg may also be used for diagnosis of chronic exertional compartment syndrome of the foot. PMID- 11912100 TI - Endoscopically assisted fasciotomy: description of technique and in vitro assessment of lower-leg compartment decompression. AB - We describe a reliable method of endoscopically assisted fasciotomy for treating chronic exertional compartment syndrome in the lower leg and for assessing compartment decompression in an in vitro model. Endoscopically assisted fasciotomy was performed in the anterior and lateral compartments of 14 matched, fresh-frozen, through-the-knee amputation specimens using a 30 degrees endoscope. A one-incision technique used in 4 specimens failed to provide complete visualization, and a two-incision technique was then performed in 10 specimens. After decompression, the skin and subcutaneous tissues were removed to assess adequacy of release, nerve decompression, anatomic course of the superficial peroneal nerve, and potential complications. Endoscopic visualization of the fascial layer and subcutaneous neurovascular structures permitted consistent compartment decompression. Fascial release, expressed as a percentage of length from the proximal origin of the fascia to the inferior retinaculum, was 99.8% (range, 98.4% to 100%) for the anterior compartment and 96.4% (range, 82% to 100%) for the lateral compartment. There were no retained fascial bands, unrecognized fascial defects, or neurovascular injuries. Optimal visualization with endoscopically assisted fasciotomy may improve clinical outcome through 1) reliable compartment decompression, 2) identification of fascial defects, 3) decompression of nerve branches at the fascial foramen, and 4) reduction of iatrogenic risk to neurovascular and muscular structures. PMID- 11912101 TI - Avulsion fracture at the femoral attachment of the anterior cruciate ligament after intercondylar eminence fracture of the tibia. PMID- 11912102 TI - Acute paraspinal muscle compartment syndrome treated with surgical decompression: a case report. PMID- 11912103 TI - Achilles tendon disorders in athletes. AB - Achilles tendon disorders are among the more common maladies seen by sports medicine physicians. Understanding the anatomy and biomechanics of the Achilles tendon and contiguous structures is essential to the diagnosis and treatment of Achilles tendon overuse injuries. Posterior heel pain is multifactorial and includes paratenonitis, tendinosis, tendinosis with partial rupture, insertional tendinitis, retrocalcaneal bursitis, and subcutaneous tendo-Achilles bursitis. Each of these entities is distinct, but they often occur in combination. Although most cases of this disorder are successfully treated nonoperatively, a small subgroup of recalcitrant cases may benefit from surgical intervention. Complete ruptures in active, athletic persons should be treated operatively in most cases and result in predictably good outcomes. There may be some cases that escape early recognition and require a reconstructive procedure to salvage a potentially severe functional deficit. PMID- 11912104 TI - The meniscal pseudocyst--a clinical sign of a torn meniscus. PMID- 11912105 TI - Toxicity of CSF in motor neurone disease: a potential route to neuroprotection. PMID- 11912106 TI - The genetics of primary dystonias and related disorders. AB - Dystonias are a heterogeneous group of disorders which are known to have a strong inherited basis. This review details recent advances in our understanding of the genetic basis of dystonias, including the primary dystonias, the 'dystonia-plus' syndromes and heredodegenerative disorders. The review focuses particularly on clinical and genetic features and molecular mechanisms. Conditions discussed in detail include idiopathic torsion dystonia (DYT1), focal dystonias (DYT7) and mixed dystonias (DYT6 and DYT13), dopa-responsive dystonia, myoclonus dystonia, rapid-onset dystonia parkinsonism, Fahr disease, Aicardi-Goutieres syndrome, Hallervorden-Spatz syndrome, X-linked dystonia parkinsonism, deafness-dystonia syndrome, mitochondrial dystonias, neuroacanthocytosis and the paroxysmal dystonias/dyskinesias. PMID- 11912107 TI - Minocycline prevents neurotoxicity induced by cerebrospinal fluid from patients with motor neurone disease. AB - CSF from patients with motor neurone disease (MND) has been reported to be toxic to cultured primary neurones. We found that CSF from MND patients homozygous for the D90A CuZn-superoxide dismutase (CuZn-SOD) mutation, patients with sporadic MND and patients with familial MND without CuZn-SOD mutations significantly increased apoptosis and reduced phosphorylation of neurofilaments in cultured spinal cord neurones when compared with the effects of CSF from patients with other neurological diseases. Exposure of spinal cord cultures to MND CSF also triggered microglial activation. The toxicity of MND CSF was independent of the presence of the CuZn-SOD mutation, and it did not correlate with gelatinase activity or the presence of immunoglobulin G autoantibodies in the CSF. The concentrations of glutamate, aspartate and glycine in MND CSF were not elevated. Antagonists of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid/kainate receptors prevented the toxic CSF-induced neuronal death but not microglial activation, whereas minocycline, a tetracycline derivative with anti-inflammatory potential independent of antimicrobial activity, reduced both the apoptotic neuronal death and microglial activation. We conclude that the cytotoxic action of CSF is prevalent in all MND cases and that microglia may mediate the toxicity of CSF by releasing excitotoxicity-enhancing factors. PMID- 11912108 TI - Inherited frontotemporal dementia in nine British families associated with intronic mutations in the tau gene. AB - Genetic screening of 171 patients with frontotemporal lobar degeneration disclosed 14 patients, across nine pedigrees, with mutations in the intron to exon 10 in the tau gene, a region regulating the splicing of exon 10 via a stem loop mechanism. Thirteen of these patients had the +16 splice site mutation and one had the +13 splice site mutation. Affected members of all nine families presented with changes in behaviour and social conduct that were prototypical of frontotemporal dementia (FTD). In all patients with the +16 splice site mutation, the behavioural profile was characterized by disinhibition, restless overactivity, a fatuous affect, puerile behaviour and verbal and motor stereotypies. The single patient with the +13 mutation presented a contrasting picture of apathy and inertia. In addition, all patients had evidence of semantic loss. Pathologically, five of the six patients so far autopsied shared frontotemporal atrophy with involvement of the substantia nigra. The underlying histology was that of microvacuolar-type cortical degeneration with a few swollen cells. Tau pathology was widespread throughout the brain and present in neurones and glial cells, mostly in the frontal and temporal cortical regions. This was in the form of neurofibrillary tangles and amorphous tau deposits (pre-tangles); Pick bodies were not observed. Ultrastructurally, the tau filaments had a twisted, ribbon-like morphology distinct from the paired helical filaments of Alzheimer's disease. One patient died from an unrelated illness whilst in the early clinical stages of FTD. In this patient, cortical microvacuolar and astrocytic changes were absent, though there were scattered neurones and glial cells, immunoreactive to tau, throughout the cortical and subcortical regions. The disease process underlying the neurodegeneration within these inherited forms of FTD may therefore stem directly from early, primary alterations in the function of tau. All eight families with the +16 mutation seem to be part of a common extended pedigree, possibly originating from a founder member residing within the North Wales region of Great Britain. PMID- 11912109 TI - Theory of mind in patients with frontal variant frontotemporal dementia and Alzheimer's disease: theoretical and practical implications. AB - A key aspect of social cognition is the ability to infer other people's mental states, thoughts and feelings; referred to as 'theory of mind' (ToM). We tested the hypothesis that the changes in personality and behaviour seen in frontal variant frontotemporal dementia (fvFTD) may reflect impairment in this cognitive domain. Tests of ToM, executive and general neuropsychological ability were given to 19 fvFTD patients, a comparison group of Alzheimer's disease patients (n = 12) and matched healthy controls (n = 16). Neuropsychiatric assessment was undertaken using the Neuropsychiatric Inventory (NPI). Patients with fvFTD were impaired on all tests of ToM (first-order false belief; second-order false belief; faux pas detection; and Reading the Mind in the Eyes), but had no difficulty with control questions designed to test general comprehension and memory. By contrast, the Alzheimer's disease group failed only one ToM task (second-order false belief), which places heavy demands on working memory. Performance on the faux pas test revealed a double dissociation, with the fvFTD group showing deficits on ToM based questions and the Alzheimer's disease group failing memory-based questions only. Rank order of the fvFTD patients according to the magnitude of impairment on tests of ToM and their degree of frontal atrophy showed a striking concordance between ToM performances and ventromedial frontal damage. There was a significant correlation between the NPI score and more sophisticated tests of ToM in the fvFTD group. This study supports the hypothesis that patients with fvFTD, but not those with Alzheimer's disease, are impaired on tests of ToM, and may explain some of the abnormalities in interpersonal behaviour that characterize fvFTD. PMID- 11912110 TI - Hypertension and cerebral white matter lesions in a prospective cohort study. AB - White matter lesions are frequently found on cerebral MRI scans of elderly people and are thought to be important in the pathogenesis of dementia. Hyper tension has been associated with the presence of white matter lesions but this has been investigated almost exclusively in cross-sectional studies. We studied prospectively the association of these lesions with the duration and treatment of hypertension. We randomly sampled 1077 subjects aged between 60 and 90 years from two prospective population-based studies. One-half of the study subjects had their blood pressure measured between 1975 and 1978 and the other half between 1990 and 1993. All subjects underwent 1.5 T MRI scanning; white matter lesions in the subcortical and periventricular regions were rated separately. Subjects with hypertension had increased rates of both types of white matter lesion. Duration of hypertension was associated with both periventricular and subcortical white matter lesions. This relationship was influenced strongly by age. For participants with >20 years of hypertension and aged between 60 and 70 years at the time of follow-up, the relative risks for subcortical and periventricular white matter lesions were 24.3 [95% confidence interval (CI) 5.1-114.8] and 15.8 (95% CI 3.4-73.5), respectively, compared with normotensive subjects. Subjects with successfully treated hypertension had only moderately increased rates of subcortical white matter lesions and periventricular white matter lesions (relative risk 3.3, 95% CI 1.3-8.4 and 2.6, 95% CI 1.0-6.8, respectively) compared with normotensive subjects. For poorly controlled hypertensives, these relative risks were 8.4 (95% CI 3.1-22.6) and 5.8 (95% CI 2.1-16.0), respectively. In conclusion, we found a relationship between long-standing hypertension and the presence of white matter lesions. Our findings are consistent with the view that effective treatment may reduce the rates of both types of white matter lesion. Adequate treatment of hypertension may therefore prevent white matter lesions and the associated cognitive decline. PMID- 11912111 TI - Analysis of fMRI and finger tracking training in subjects with chronic stroke. AB - Hand movement recovery and cortical reorganization were studied in 10 subjects with chronic stroke using functional MRI (fMRI) before and after training with an intensive finger movement tracking programme. Subjects were assigned randomly to a treatment or control group. The treatment group received 18-20 sessions of finger tracking training using target waveforms under variable conditions. The control group crossed over to receive the same treatment after the control period. For comparison with a healthy population, nine well elderly females were also studied; however, the well elderly controls did not cross over after the control period. The dependent variables consisted of a Box and Block score to measure prehensile ability (subjects with stroke only), a tracking accuracy score and quantification of active cortical areas using fMRI. For the tracking tests, the subjects tracked a sine wave target on a computer screen with extension and flexion movements of the paretic index finger. Functional brain images were collected from the frontal and parietal lobes of the subject with a 4 tesla magnet. Areas of interest included the sensorimotor cortex (SMC), primary motor area (M1), primary sensory area (S1), premotor cortex (PMC) and supplementary motor area (SMA). Comparison between all subjects with stroke and all well elderly subjects at pre-test was analysed with two-sample t-tests. Change from pre-test to post-test within subjects was analysed with paired t-tests. Statistical significance was set at P < 0.05. Stroke treatment subjects demonstrated significant improvement in tracking accuracy, whereas stroke control subjects did not until after crossover treatment. At pre-test, the cortical activation in the subjects with stroke was predominantly ipsilateral to the performing hand, whereas in the well elderly subjects it was contralateral. Activation for the stroke treatment group following training switched to contralateral in SMC, M1, S1 and PMC. The stroke control group's activation remained ipsilateral after the control period, but switched to contralateral after crossover to receive treatment. All well elderly subjects maintained predominantly contralateral activation throughout. Transfer of skill to functional activity was shown in significantly improved Box and Block scores for the stroke treatment group, with no such improvement in the stroke control group until after crossover. We concluded that individuals with chronic stroke receiving intensive tracking training showed improved tracking accuracy and grasp and release function, and that these improvements were accompanied by brain reorganization. PMID- 11912112 TI - Frontal atrophy correlates with behavioural changes in progressive supranuclear palsy. AB - Regional brain volumes were measured in 21 patients with progressive supranuclear palsy (PSP), 17 patients with Parkinson's disease and 23 controls using 3D MRI based volumetry. Cortical, subcortical and ventricular volume measures were correlated with global indices of motor disability and cognitive disturbance. All MRI measures, including hippocampal volume, were preserved in Parkinson's disease. Patients with PSP could be distinguished from both Parkinson's disease and controls by whole brain volume loss, ventricular dilatation and disproportionate atrophy of the frontal cortex. Caudate nucleus volume loss additionally differentiated PSP from controls, but was modest in severity and proportionate to whole brain volume loss. The present study identifies disease specific differences in the topography of brain atrophy between PSP and Parkinson's disease, and has potential implications for the in vivo radiological differentiation of these two disorders. In PSP, the variance in frontal grey matter volume related to measures of behavioural disturbance, confirming the use of behavioural tests for ante-mortem case differentiation and suggesting that intrinsic cortical deficits contribute to these clinical disturbances. PMID- 11912113 TI - Guadeloupean parkinsonism: a cluster of progressive supranuclear palsy-like tauopathy. AB - An unusually high frequency of atypical Parkinson syndrome has been delineated over the last 5 years in the French West Indies. Postural instability with early falls, prominent frontal lobe dysfunction and pseudo-bulbar palsy were common and three-quarters of the patients were L-dopa unresponsive. One-third of all patients seen had probable progressive supranuclear palsy (PSP). This new focus of atypical parkinsonism is reminiscent of the one described in Guam and may be linked to exposure to tropical plants containing mitochondrial complex I inhibitors (quinolines, acetogenins, rotenoids). Two hundred and twenty consecutive patients with Parkinson's syndrome seen by the neurology service at Pointe a Pitre, Guadeloupe University Hospital were studied. Currently accepted operational clinical criteria for Parkinson's syndromes were applied. The pathological findings of three patients who came to autopsy are reported. Fifty eight patients had probable PSP, 96 had undetermined parkinsonism and 50 had Parkinson's disease, 15 had amyotrophic lateral sclerosis with parkinsonism and one had probable multiple system atrophy. All three PSP patients in whom post mortem study was performed had early postural instability, gaze palsy and parkinsonian symptoms, followed by a frontolimbic dementia and corticobulbar signs. Neuropathological examination showed an accumulation of tau proteins, predominating in the midbrain. There was an exceptionally large accumulation of neuropil threads in Case 1. Biochemical studies detected a major doublet of pathological tau at 64 and 69 kDa in brain tissue homogenates. All cases were homozygous for the H1 tau haplotype, but no mutation of the tau gene was observed. Clinical, neuropathological and biochemical features were compatible with the diagnosis of PSP, although some unusual pathological features were noted in Case 1. A cluster of cases presenting with atypical parkinsonism is reported. Guadeloupean parkinsonism may prove to be a tauopathy identical or closely related to PSP. PMID- 11912114 TI - Contrast-enhanced MRI in acute optic neuritis: relationship to visual performance. AB - The location and extent of an abnormal signal on MRI of the optic nerve affected by optic neuritis are said to correlate with the severity of initial visual loss and recovery. We used gadolinium-enhanced fat-suppressed MRI to show abnormal enhancement of the optic nerve to determine the sensitivity of this modality in acute optic neuritis and whether the abnormal enhancement correlates with presenting visual deficits or recovery. A total of 107 patients, 93 with follow up (68 steroid treated), were included; 101 patients had enhancement of the affected optic nerve and no unaffected nerve enhanced. The baseline visual performance was similar between nerves with and without enhancement. Optic nerves with enhancement in the optic canal had poorer colour vision (P = 0.04) and nerves with all segments involved had worse threshold perimetry (P = 0.001) and colour vision (P = 0.008). Nerves with enhancement >10 mm had worse threshold perimetry (P = 0.004), while nerves with enhancing segments >17 mm had poorer baseline visual acuity (P = 0.02), threshold perimetry (P = 0.009) and colour vision (P = 0.01). For all parameters of vision, recovery was similar regardless of location or length of abnormal enhancement. Abnormal contrast enhancement of the optic nerve is a sensitive (94%) finding in acute optic neuritis and is absent in unaffected or previously affected optic nerves. Although lesions involving the canal or longer segments of optic nerve have worse starting vision, the location and length of enhancement are not predictive of recovery. PMID- 11912115 TI - Chemokines and chemokine receptors in inflammatory demyelinating neuropathies: a central role for IP-10. AB - Inflammatory cell recruitment is an important step in the pathogenesis of autoimmune demyelinating diseases of the PNS. Chemokines might play a critical role in promoting leucocyte entry into the nervous system during immune-mediated inflammation. Here, we report the expression pattern of the chemokine receptors CCR-1, CCR-2, CCR-4, CCR-5 and CXCR-3 in sural nerve biopsies obtained from patients with classical Guillain-Barre syndrome (acute inflammatory demyelinating polyradiculoneuropathy), chronic inflammatory demyelinating polyradiculoneuropathy and various non-inflammatory neuropathies. A consistent chemokine receptor expression pattern was immunohistochemically detected in inflammatory demyelinating neuropathies and quantitation of labelled mononuclear cells revealed significantly elevated cell counts compared with controls. CCR-1 and CCR-5 were primarily expressed by endoneurial macrophages, whereas CCR-2, CCR 4 and CXCR-3 could be localized to invading T lymphocytes. Quantitative analysis revealed that CXCR-3 was expressed at highest numbers by infiltrating T cells compared with the other receptors. Thus, expression and distribution of CXCR-3 suggest a specific role of this receptor in chemokine-mediated lymphocyte traffic into the inflamed PNS tissue. Therefore, we further analysed the expression of its ligands interferon-gamma-inducible protein of 10 kDa (IP-10) and monokine induced by interferon-gamma (Mig). Significantly increased levels of IP-10 could be measured in the CSF of patients with inflammatory neuropathies, whereas no differences were observable for Mig. In situ hybridization for IP-10 mRNA mirrored the distribution of the cognate receptor within the inflamed PNS, and delineated endothelial cells as the primary cellular source of IP-10. Our results imply a pathogenic role for specific chemokine receptors and IP-10 in the genesis of inflammatory demyelinating neuropathies. PMID- 11912116 TI - Enhanced inactivation and pH sensitivity of Na(+) channel mutations causing hypokalaemic periodic paralysis type II. AB - Hypokalaemic periodic paralysis (hypoPP) is a dominantly inherited muscle disorder characterized by episodes of flaccid weakness. Previous genetic studies revealed mutations in the voltage-gated calcium channel alpha1-subunit (CACNA1S gene) in families with hypoPP (type I). Electrophysiological studies on these mutants in different expression systems could not explain the pathophysiology of the disease. In addition, several mutations (Arg669His, Arg672His, Arg672Gly and Arg672Ser) in the voltage sensor of the skeletal muscle sodium channel alpha subunit (SCN4A gene) have been found in families with hypoPP (type II). For Arg672Gly/His a fast inactivation defect was described, and for Arg669His an impairment of slow inactivation was reported. Except for the substitution for serine, we have now expressed all mutants in a human cell-line and studied them electrophysiologically. Patch-clamp recordings show an enhanced fast inactivation for all three mutations, whereas two of them reveal enhanced slow inactivation. This may reduce the number of functional sodium channels at resting membrane potential and contribute to the long-lasting periods of paralysis experienced by hypoPP patients. The gating of both histidine mutants (Arg669His, Arg672His) can be modulated by changes of extra- or intracellular pH. The inactivation defects of Arg669His and Arg672His can be alleviated by low pH to a significant degree, suggesting that the decrease of pH in muscle cells (e.g. during muscle work) might lead to an auto-compensation of functional defects. This may explain a delay or prevention of paralytic attacks in patients by slight physical activity. Moreover, the histidine residues may be the target for a potential therapeutic action by acetazolamide. PMID- 11912118 TI - The accuracy of diagnosis of parkinsonian syndromes in a specialist movement disorder service. AB - We have reviewed the clinical and pathological diagnoses of 143 cases of parkinsonism seen by neurologists associated with the movement disorders service at The National Hospital for Neurology and Neurosurgery in London who came to neuropathological examination at the United Kingdom Parkinson's Disease Society Brain Research Centre, over a 10-year period between 1990 and the end of 1999. Seventy-three (47 male, 26 female) cases were diagnosed as having idiopathic Parkinson's disease (IPD) and 70 (42 male, 28 female) as having another parkinsonian syndrome. The positive predictive value of the clinical diagnosis for the whole group was 85.3%, with 122 cases correctly clinically diagnosed. The positive predictive value of the clinical diagnosis of IPD was extremely high, at 98.6% (72 out of 73), while for the other parkinsonian syndromes it was 71.4% (50 out of 70). The positive predictive values of a clinical diagnosis of multiple system atrophy (MSA) and progressive supranuclear palsy (PSP) were 85.7 (30 out of 35) and 80% (16 out of 20), respectively. The sensitivity for IPD was 91.1%, due to seven false-negative cases, with 72 of the 79 pathologically established cases being diagnosed in life. For MSA, the sensitivity was 88.2% (30 out of 34), and for PSP it was 84.2% (16 out of 19). The diagnostic accuracy for IPD, MSA and PSP was higher than most previous prospective clinicopathological series and studies using the retrospective application of clinical diagnostic criteria. The seven false-negative cases of IPD suggest a broader clinical picture of disease than previously thought acceptable. This study implies that neurologists with particular expertise in the field of movement disorders may be using a method of pattern recognition for diagnosis which goes beyond that inherent in any formal set of diagnostic criteria. PMID- 11912117 TI - Post-ischaemic long-term synaptic potentiation in the striatum: a putative mechanism for cell type-specific vulnerability. AB - In the present in vitro study of rat brain, we report that transient oxygen and glucose deprivation (in vitro ischaemia) induced a post-ischaemic long-term synaptic potentiation (i-LTP) at corticostriatal synapses. We compared the physiological and pharmacological characteristics of this pathological form of synaptic plasticity with those of LTP induced by tetanic stimulation of corticostriatal fibres (t-LTP), which is thought to represent a cellular substrate of learning and memory. Activation of N-methyl-D-aspartate (NMDA) receptors was required for the induction of both forms of synaptic plasticity. The intraneuronal injection of the calcium chelator BAPTA [bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetate] and inhibitors of the mitogen activated protein kinase pathway blocked both forms of synaptic plasticity. However, while t-LTP showed input specificity, i-LTP occurred also at synaptic pathways inactive during the ischaemic period. In addition, scopolamine, a muscarinic receptor antagonist, prevented the induction of t-LTP but not of i LTP, indicating that endogenous acetylcholine is required for physiological but not for pathological synaptic potentiation. Finally, we found that striatal cholinergic interneurones, which are resistant to in vivo ischaemia, do not express i-LTP while they express t-LTP. We suggest that i-LTP represents a pathological form of synaptic plasticity that may account for the cell type specific vulnerability observed in striatal spiny neurones following ischaemia and energy deprivation. PMID- 11912120 TI - Deficits and recovery of head and trunk orientation and stabilization after unilateral vestibular loss. AB - The aim of the study was to analyse changes in the orientation and stabilization of the head and trunk and their recovery after complete unilateral loss of vestibular information in humans. The ability of nine Meniere's patients to orient and stabilize their heads and trunks in space was investigated during a simple dynamic task of knee-bends and compared with the performance of 10 healthy subjects. Patients' performance was recorded before unilateral vestibular neurotomy (UVN) and during the time-course of recovery (1 week, 1 month, 3 months). Experiments were performed both in eyes open (EO) and eyes closed (EC) conditions to evaluate the role of visual cues in the recovery process. Head and trunk mean angular position (orientation) and mean maximal angular rotation (stabilization) in the roll plane and the yaw plane were recorded using a video motion analysis system. The results indicate that, in the acute stage after UVN (1 week), patients exhibit marked impairments in head and trunk orientation in both visual conditions. In the EC condition, head and trunk were deviated towards the operated side in the roll plane and the yaw plane. Head and trunk stabilization in space was impaired in the roll plane and associated with increased stabilization of the head on the shoulders. Interestingly, vision caused a complete inversion of the orientation pattern, with head and trunk rotations towards the intact side in the roll plane and the yaw plane. Relative to darkness, vision also reduced head and trunk oscillations. Recovery from abnormal head orientation in the light and impaired head stability in both visual conditions was achieved within 1 month and 3 months after UVN, respectively. However, head and trunk orientation in the dark and trunk stabilization in the roll plane remained uncompensated 3 months post-lesion. These results suggest that unilateral vestibular loss leads to a postural syndrome similar to that described previously for various animal species. They confirm the necessity of vestibular inputs for properly stabilizing head and trunk during self-generated displacements in healthy subjects. They also support the notion that vestibular compensation relies on visual cues whose substitution role gradually decreases after UVN. PMID- 11912121 TI - Abnormal cortical mechanisms of voluntary muscle relaxation in patients with writer's cramp: an fMRI study. AB - Although it is hypothesized that there is abnormal motor inhibition in patients with dystonia, the question remains as to whether the mechanism related to motor inhibition is specifically impaired. The objective of the present study was to clarify the possible abnormalities of the mechanisms underlying voluntary muscle relaxation during motor preparation and execution in patients with writer's cramp, using event-related functional MRI. Eight patients with writer's cramp and 12 age-matched control subjects participated in the study. Two motor tasks were employed as an experimental paradigm. In the relaxation task, subjects were asked to hold their right wrist in the horizontal plane by maintaining moderate contraction of wrist extensor muscles in the premotor phase; they relaxed those muscles voluntarily just once during each fMRI scanning session. In the contraction task, subjects extended the right wrist voluntarily from the same premotor state as for the relaxation task. Five axial images covering the primary sensorimotor cortex (SMC) and supplementary motor area (SMA) were obtained once every second. Activated volumes in the left SMC and the SMA were significantly reduced in patients for both muscle relaxation and contraction tasks. These data suggest that there is impaired activation in both SMC and SMA in voluntary muscle relaxation and contraction in patients with writer's cramp. This implies that abnormalities of both inhibitory and excitatory mechanisms in motor cortices might play a role in the pathophysiology of focal dystonia. PMID- 11912119 TI - Force overflow and levodopa-induced dyskinesias in Parkinson's disease. AB - We assessed force coordination of the hand in Parkinson's disease and its relationship to motor complications of levodopa therapy, particularly to levodopa induced dyskinesias (LID). We studied two groups of Parkinson's disease patients with (Parkinson's disease + LID, n = 23) and without levodopa-induced dyskinesias (Parkinson's disease - LID, n = 10), and age-matched healthy controls. The motor score of the Unified Parkinson's Disease Rating Scale, a dyskinesia score and force in a grip-lift paradigm were assessed ON and OFF levodopa. A pathological increase of forces was seen in ON-state in Parkinson's disease + LID only. In Parkinson's disease + LID, the force involved in pressing down the object before lifting was significantly increased by levodopa (by 61%, P < 0.05). An overshooting of peak grip force by 51% (P < 0.05) and of static grip force by 45% (P < 0.01) was observed in the ON- compared with the OFF-drug condition. In contrast, no excessive force was found in Parkinson's disease - LID. Peak grip force in ON-state was 140% (P < 0.05) higher in Parkinson's disease + LID than in Parkinson's disease - LID, while static grip force was increased by 138% (P < 0.01) between groups. Severity of peak-dose dyskinesias was strongly correlated with grip force in ON-state (r = 0.79 with peak force, P < 0.01). No correlation was observed between forces and the motor score as well as with the daily dose of dopaminergic medication. Force excess was only observed in patients with LID and motor fluctuations. A close relationship was seen between the overshooting of forces and dyskinesias in the ON-drug condition. We postulate that both LID and grip force excess share common pathophysiological mechanisms related to motor fluctuations. PMID- 11912123 TI - Effects of repetitive transcranial magnetic stimulation on visual evoked potentials in migraine. AB - Between attacks, migraine patients are characterized by potentiation instead of habituation of stimulation-evoked cortical responses. It is debated whether this is due to increased or decreased cortical excitability. We have studied the changes in visual cortex excitability by recording pattern-reversal visual evoked potentials (PR-VEP) after low- and high-frequency repetitive transcranial magnetic stimulation (rTMS), known respectively for their inhibitory and excitatory effect on the cortex. In 30 patients (20 migraine without, 10 with aura) and 24 healthy volunteers, rTMS of the occipital cortex was performed with a focal figure-of-eight magnetic coil (Magstim). Nine hundred pulses were delivered randomly at 1 or 10 Hz in two separate sessions. Stimulus intensity was set to the phosphene threshold or to 110% of the motor threshold if no phosphenes were elicited. Before and after rTMS, PR-VEP were averaged sequentially in six blocks of 100zztieresponses during uninterrupted 3.1 Hz stimulation. In healthy volunteers, PR-VEP amplitude was significantly decreased in the first block after 1 Hz rTMS and the habituation normally found in successive blocks after sustained stimulation was significantly attenuated. In migraine patients, 10 Hz rTMS was followed by a significant increase of first block PR-VEP amplitude and by a reversal to normal habituation of the potentiation (or dishabituation) characteristic of the disorder. This effect was similar in both forms of migraine and lasted for at least 9 min. There were no significant changes of PR-VEP amplitudes after 1 Hz rTMS in migraineurs and after 10 Hz rTMS in healthy volunteers, nor after sham stimulation. The recovery of a normal PR-VEP habituation pattern after high-frequency rTMS is probably due to activation of the visual cortex and the dishabituation in healthy volunteers to cortical inhibition. We conclude, therefore, that the deficient interictal PR-VEP habituation in migraine is due to a reduced, and not to an increased, pre activation excitability level of the visual cortex. PMID- 11912122 TI - Anti-myelin-associated glycoprotein antibodies alter neurofilament spacing. AB - Axon calibre is crucial to efficient impulse transmission in the peripheral nervous system. Neurofilament numbers determine gross axonal diameter, but intra axonal distribution depends on the phosphorylation status of neurofilament sidearms. Myelin-associated glycoprotein (MAG) has been implicated in the signalling cascade controlling neurofilament phosphorylation and hence in the control of axon calibre. In an electron microscopic morphometric study we measured nearest neighbour neurofilament distances (NNND) in the axons of sural nerves from patients with anti-MAG paraproteinaemic neuropathies and compared these with normal human sural nerves and those from patients with Guillain-Barre syndrome or chronic inflammatory demyelinating polyradiculoneuropathy. Axon calibre was similar in all groups. In normal human sural nerves, axonal NNND was correlated with axonal diameter (r = 0.56). In diseased axons this correlation did not exist. The NNND was significantly reduced in demyelinated axons (30.5+/ 2.2 nm) and those with widely spaced myelin (28.9+/-1.3 nm) from patients with anti-MAG antibodies compared with normal axons from normal patients (39.8+/- 3.2 nm) or those with demyelinating neuropathy (35.8+/-4.6 nm). This reinforces the hypothesis that MAG is involved in the control of neurofilament spacing through sidearm phosphorylation and demonstrates a MAG-mediated pathogenic effect of the anti-MAG antibody in peripheral nerves. PMID- 11912125 TI - Doxycycline decreases tumor burden in a bone metastasis model of human breast cancer. AB - Bone is one of the most frequent sites for metastasis in breast cancer patients,often resulting in significant clinical morbidity and mortality. Increased matrix metalloproteinase (MMP) activity of tumor cells correlates with a higher invasive and metastatic potential. Members of the tetracycline family of antibiotics, including doxycycline, have potential treatment value for bone metastasis; they inhibit cancer cell proliferation, and they are also potent MMP inhibitors and are highly osteotropic. Doxycycline treatment in an experimental bone metastasis mouse model of human breast cancer MDA-MB-231 cells resulted in a 70% reduction in total tumor burden when compared with placebo control animals. In tumor-bearing animals, the amount of doxycycline incorporated into the radius/ulna as assessed by ELISA was lower than in non-tumor-bearing animals. In doxycycline-treated mice, bone formation was significantly enhanced as determined by increased numbers of osteoblasts, osteoid surface, and volume, whereas a decrease in bone resorption was also observed. Doxycycline treatment may be beneficial for breast cancer patients with or at risk for osteolytic bone metastasis; it greatly reduces tumor burden and could also compensate for the increased bone resorption associated with the disease. PMID- 11912124 TI - Requirement of BAX for TRAIL/Apo2L-induced apoptosis of colorectal cancers: synergism with sulindac-mediated inhibition of Bcl-x(L). AB - The cornerstone of the systemic treatment of advanced colorectal cancer is 5 fluorouracil.However, 5-fluorouracil-induced apoptosis is dependent on p53, a tumor suppressor gene that is lost or inactivated in at least 85% of human colorectal cancers. Here we show that tumor necrosis factor-related apoptosis inducing ligand (TRAIL)/Apo2L triggers caspase-8-mediated truncation of BID, mitochondrial activation of caspase-9, and apoptosis in both p53(+/+) or p53(-/-) isogenic HCT116 colorectal cancer cells. TRAIL/Apo2L also sensitizes both p53(+/+) or p53(-/-) colorectal cancer cells to ionizing radiation. In contrast, we find that TRAIL/Apo2L fails to activate caspase-9 or induce apoptosis in isogenic HCT116 colorectal cancer cells that are deficient in BAX, a proapoptotic gene that is mutated in >50% of colorectal cancers of the microsatellite mutator phenotype. Loss of BAX also renders colorectal cancer cells resistant to TRAIL/Apo2L-mediated radiosensitization. We additionally demonstrate that TRAIL/Apo2L-induced death of p53(+/+)- or p53(-/-)- BAX-proficient but not BAX deficient colorectal cancer cells is augmented by reducing nuclear factor-kappaB dependent expression of Bcl-x(L) with either a peptide that disrupts the inhibitor of kappaB kinase complex or the nonsteroidal anti-inflammatory drug, sulindac sulfide. These results indicate that the combination of TRAIL/Apo2L with either irradiation or sulindac may be highly effective against both p53 proficient and p53-deficient colorectal cancers; however, BAX-deficient tumors may evade elimination by TRAIL/Apo2L-based regimens. Our findings may aid the development and genotype-specific application of TRAIL/Apo2L-based combinatorial regimens for the treatment of colorectal cancers. PMID- 11912126 TI - Selective depletion of human DNA-methyltransferase DNMT1 proteins by sulfonate derived methylating agents. AB - 5-Methylcytosine residues in the DNA (DNA methylation) are formed from the transfer of the methyl group from S-adenosylmethionine to the C-5 position of cytosine by the DNA-(cytosine-5) methyltransferases (DNMTs). Although regional hypermethylation and global hypomethylation of the genome are commonly observed in neoplastic cells, how these aberrant methylation patterns occur remains unestablished. We report here that sulfonate-derived methylating agents, unlike N methylnitrosourea or iodomethane, are potent in depleting DNMT1 proteins in human cells, in addition to their DNA-damaging properties. Their effects on cellular DNMT1 are time and dosage dependent but independent of cell type. Unlike gamma irradiation, these agents apparently do not activate the p53/p21(WAF1) DNA damage response pathway to deplete the DNMT1 proteins because cells with wild-type, mutated, or inactivated p53 behave similarly. However, cell cycle analysis and protease assay studies strongly suggest that methylmethanesulfonate may activate a cellular protease to degrade DNMT1. These results explain why reported observations on the effect of alkylating agents on DNMT1 activities in human cells vary significantly and provide a crucial link to understand the mechanism behind genomic hypomethylation. PMID- 11912127 TI - An ATM-independent S-phase checkpoint response involves CHK1 pathway. AB - After exposure to genotoxic stress, proliferating cells actively slow down the DNA replication through a S-phase checkpoint to provide time for repair. We report that in addition to the ataxia-telangiectasia mutated (ATM)-dependent pathway that controls the fast response, there is an ATM-independent pathway that controls the slow response to regulate the S-phase checkpoint after ionizing radiation in mammalian cells. The slow response of S-phase checkpoint, which is resistant to wortmannin, sensitive to caffeine and UCN-01, and related to cyclin dependent kinase phosphorylation, is much stronger in CHK1 overexpressed cells, and it could be abolished by Chk1 antisense oligonucleotides. These results provide evidence that the ATM-independent slow response of S-phase checkpoint involves CHK1 pathway. PMID- 11912128 TI - Generation of effective antitumor vaccines using photodynamic therapy. AB - Preclinical studies have shown that photodynamic therapy (PDT) of tumors augments the host antitumor immune response. However, the role of the PDT effect on tumor cells as opposed to the host tissues has not been determined. To test the contribution of the direct effects of PDT on tumor cells to the enhanced antitumor response by the host, we examined the immunogenicity of PDT-generated murine tumor cell lysates in a preclinical vaccine model. We found that the PDT generated tumor cell lysates were potent vaccines and that PDT-generated vaccines are more effective than other modes of creating whole tumor vaccines, i.e., UV or ionizing irradiation, and unlike other traditional vaccines, PDT vaccines do not require coadministration of an adjuvant to be effective. PDT vaccines are tumor specific and appear to induce a cytotoxic T-cell response. We have demonstrated that although both UV and PDT-generated tumor cell lysates are able to induce phenotypic DC maturation, only PDT-generated lysates are able to activate DCs to express IL-12, which is critical to the development of a cellular immune response. Our results show that PDT effects on tumor cells alone are sufficient to generate an antitumor immune response, indicating that the direct tumor effects of PDT play an important role in enhancing that host antitumor immune response. These studies also suggest that in addition to the role of PDT as a therapeutic modality, PDT-generated vaccines may have clinical potential as an adjuvant therapy. PMID- 11912129 TI - Target gene mutation profile differs between gastrointestinal and endometrial tumors with mismatch repair deficiency. AB - Mutation frequencies at 25 genes containing coding repeats were compared in colorectal, gastric, and endometrial mismatch repair-deficient (MSI-H) tumors. The overall number of mutations was significantly lower in endometrial than in gastrointestinal MSI-H cancers. Using a likelihood statistical method, target genes were divided in each tumor location into two groups likely to represent gene mutations that do or do not provide selective pressures during tumoral progression. Mutation profiles were quite similar in gastric and colorectal MSI-H cancers but were different in endometrial MSI-H tumors. Deletions in Bat-25 and Bat-26 noncoding repeats were also significantly less important in endometrial as compared with gastrointestinal MSI-H tumors. Our results show that the profile of target gene mutations in MSI-H tumors is tissue specific, with both qualitative and quantitative differences between gastrointestinal and endometrial MSI-H cancers. PMID- 11912130 TI - The SLUG zinc-finger protein represses E-cadherin in breast cancer. AB - Loss of expression of the E-cadherin cell-cell adhesion molecule is important in carcinoma development and progression. Because previous data suggest that loss of E-cadherin expression in breast carcinoma may result from a dominant transcriptional repression pathway acting on the E-cadherin proximal promoter, we pursued studies of cis sequences and transcription factors regulating E-cadherin expression in breast cancer cells. E-box elements in the E-cadherin promoter were found to play a critical negative regulatory role in E-cadherin gene transcription in breast cancer cell lines lacking E-cadherin transcription. The E box elements had a minimal role in E-cadherin transcription in breast cancer cell lines expressing E-cadherin. Two zinc-finger transcription factors known to bind E-box elements, SLUG and SNAIL, repressed E-cadherin-driven reporter gene constructs containing wild-type promoter sequences but not those with mutations in the E-box elements. Additionally, both SLUG and SNAIL repressed endogenous E cadherin expression. These findings suggest SLUG and SNAIL are potential repressors of E-cadherin transcription in carcinomas lacking E-cadherin expression. Analysis of the expression patterns of SLUG, SNAIL, and E-cadherin in breast cancer cell lines demonstrated that expression of SLUG was strongly correlated with loss of E-cadherin transcripts. Taken together, the data indicate the E-box elements in the proximal E-cadherin promoter are critical in transcriptional repression of the E-cadherin gene, and SLUG is a likely in vivo repressor of E-cadherin in breast cancer. PMID- 11912131 TI - Osteoprotegerin (OPG) is a survival factor for human prostate cancer cells. AB - Factors that aid survival of prostate cancer cells in the presence of the various categories of cytotoxic cytokines present in tumors in vivo are largely unknown. Osteoprotegerin (OPG) is a decoy receptor for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) that inhibits TRAIL-induced apoptosis. In relation to this activity, we hypothesized that the ability to produce OPG by prostate cancer cells would confer a survival advantage on these cells. In this study we have demonstrated that high levels of OPG are produced by the hormone insensitive prostate cancer cell lines PC3 and Du145, whereas the hormone sensitive cell line LNCaP produced 10-20-fold less OPG under the same conditions. A strong negative correlation was observed between levels of endogenously produced OPG in the medium and the capacity of TRAIL to induce apoptosis in cells that produced high levels of OPG. The antiapoptotic effect of OPG was reversed by coadministration of 100-fold molar excess of receptor-activator of nuclear factor kappaB ligand, another protein that selectively binds OPG. These observations suggest that prostate cancer-derived OPG may be an important survival factor in hormone-resistant prostate cancer cells. PMID- 11912132 TI - Characterization of human UMP/CMP kinase and its phosphorylation of D- and L-form deoxycytidine analogue monophosphates. AB - Pyrimidine nucleoside monophosphate kinase [UMP/CMP kinase (UMP/CMPK);EC 2.7.4.14] plays a crucial role in the formation of UDP, CDP, and dCDP, which are required for cellular nucleic acid synthesis. Several cytidine and deoxycytidine analogues are important anticancer and antiviral drugs. These drugs require stepwise phosphorylation to their triphosphate forms to exert their therapeutic effects. The role of UMP/CMPK for the phosphorylation of nucleoside analogues has been indicated. Thus, we cloned the human UMP/CMPK gene, expressed it in Escherichia coli, and purified it to homogeneity. Its kinetic properties were determined. UMP and CMP proved to be far better substrates than dCMP. UMP/CMPK used all of the nucleoside triphosphates as phosphate donors, with ATP and dATP being the best donors and CTP being the poorest. Furthermore, UMP/CMPK was able to phosphorylate all of the deoxycytidine analogue monophosphates that we tested. The relative efficiency was as follows: arabinofuranosyl-CMP > dCMP > beta-L 2',3'-dideoxy-3'-thia-CMP > Gemcitabine monophosphate > beta-D-2',3'-dideoxy-CMP; beta-L-2',3'-dideoxy-2',3'-didehydro-5-fluoro-CMP; beta-L-2',3'-dideoxy-5-fluoro 3'-thia-CMP > beta-L-2',3'-dideoxy-CMP > beta-L-dioxolane-CMP. By comparing the relative V(max)/K(m) values of D- and L-form dideoxy-CMP, we showed that this kinase lacked stereoselectivity. Reducing agents, such as DTT, 2-mercaptoethanol, and thioredoxin, were able to activate this enzyme, suggesting that its activity may be regulated by redox potential in vivo. UMP/CMPK localized predominantly to the cytoplasm. In addition, 196-amino acid UMP/CMPK was the actual form of UMP/CMPK, rather than the 228-amino acid form as suggested before. PMID- 11912133 TI - Neurotensin induces protein kinase C-dependent protein kinase D activation and DNA synthesis in human pancreatic carcinoma cell line PANC-1. AB - Signal transduction pathways through protein kinase C (PKC) may play a significant role in DNA synthesis and proliferation of human pancreatic cancers. Treatment of human pancreatic ductal adenocarcinoma cell line PANC-1 with biologically active phorbol-12,13-dibutyrate led to striking activation of protein kinase D (PKD), a member of a novel family of serine/threonine kinases distinct from PKC isoforms. Using PANC-1 as a model system, we demonstrate that neurotensin (NT) induced a rapid and striking activation of PKD as determined by in vitro kinase assay and by in vivo phosphorylation of serines 744, 748, and 916. PKD activation induced by NT was abrogated by treatment of PANC-1 cells with PKC inhibitors GF-1 and Ro 31-8220. NT induced a rapid and transient translocation of PKD from the cytosol to the plasma membrane. Inhibiting PKC activity blocked the reverse translocation of PKD from the plasma membrane to the cytosol. Finally, we show that NT-induced DNA synthesis in PANC-1 cells is PKC dependent. Collectively our results demonstrate, for the first time, the existence of a functional PKC/PKD signaling pathway in human ductal pancreatic carcinoma cells and suggest that PKCs mediate the mitogenic signaling process initiated by NT. PMID- 11912134 TI - Enhanced skin carcinogenesis in cyclin D1-conditional transgenic mice: cyclin D1 alters keratinocyte response to calcium-induced terminal differentiation. AB - Cyclin D1 is a critical gene involved in the regulation of progression through the G(1) phase of the cell cycle, thereby contributing to cell proliferation. Gene amplification and abnormal expression of Cyclin D1 have been described in several human cancers. To understand their biological significance in skin carcinogenesis, we established Cyclin D1-conditional transgenic mice with C57BL/6J background, in which skin-specific overexpression of Cyclin D1 transgene was observed. The mice were subjected to dimethylbenz[a]anthracene complete skin carcinogenesis studies. After 40 weeks of repeated administration of dimethylbenz[a]anthracene on the skin (once a week), all of the mice with high Cyclin D1 expression had papillomas, whereas only 9.5% of the control mice without the transgene developed papillomas. Primary cultured keratinocytes with induced Cyclin D1 transgene expression showed resistance to calcium-induced terminal differentiation and continued to replicate in vitro. These results clearly provide us with direct experimental evidence that overexpression of CyclinD1 induces excessive dermal cell proliferation via the altered differentiation state of keratinocytes. The conditional transgenic mice described here provide excellent in vivo and in vitro model systems to understand the role of cyclin D1 and deregulation of the cell cycle in carcinogenesis. PMID- 11912135 TI - Nitric oxide inhibits apoptosis downstream of cytochrome C release by nitrosylating caspase 9. AB - Inhibition of the mitochondrial pathway of apoptosis has been implicated as a mechanism contributing to carcinogenesis. Chronic inflammation, which is accompanied by activation of inducible nitric oxide synthase and generation of nitric oxide (NO), is associated with cancer development in a variety of gastrointestinal diseases, including cholangiocarcinoma. Therefore, we examined the effects of NO on the mitochondrial pathway of apoptosis in human cholangiocarcinoma cell lines. Transfection with inducible NO synthase inhibited etoposide-induced apoptosis. S-Nitroso-N-acetyl-D,L-penicillamine (SNAP), a pharmacological NO donor, did not prevent mitochondrial cytochrome c release as assessed by immunoblot analysis or cellular localization of cytochrome c-green fluorescent protein. In contrast, SNAP did prevent activation of caspase 9 in etoposide-treated cells. Furthermore, SNAP also blocked caspase 9 activation in a cell-free system and reversibly inhibited catalytic activity of human recombinant caspase 9. As assessed by the Saveille reaction, immunoprecipitated procaspase 9 from SNAP-treated cells released 6-fold more NO than untreated cells, confirming that cellular procaspase 9 is susceptible to nitrosylation. In conclusion, NO inhibits apoptosis downstream of cytochrome c release by directly blocking caspase 9 activation. PMID- 11912136 TI - Retinol metabolism and lecithin:retinol acyltransferase levels are reduced in cultured human prostate cancer cells and tissue specimens. AB - Recent studies from our laboratory have indicated that the metabolism of vitamin A (retinol) to retinyl esters, carried out primarily by the enzyme lecithin:retinol acyltransferase (LRAT), is greatly reduced in human carcinoma cell lines of the oral cavity, skin, breast, and kidney as compared with their normal epithelial counterparts. These studies suggest that human carcinoma cells are retinoid-deficient relative to normal epithelial cells. In this study, we examined the metabolism of [(3)H]retinol and [(3)H]retinoic acid (RA) in human prostate cancer lines and in primary cultures of human prostate epithelial cells. Normal cells esterified all of the [(3)H]retinol added to the cultures. In contrast, all seven prostate cancer cell lines and four primary cultures derived from prostatic adenocarcinomas metabolized only trace amounts of [(3)H]retinol to [(3)H]retinyl esters. Correlated with this relative lack of esterification of [(3)H]retinol by the cancer cells was loss of expression of LRAT protein, whereas normal cells expressed abundant levels of LRAT protein by Western analysis. The metabolism of [(3)H]RA was also examined in these prostatic cells. Two of the prostate cancer tumor lines, DU 145 and PJ-1, exhibited rapid metabolism of [(3)H]RA; in contrast, the other tumor lines or primary cultures metabolized [(3)H]RA at a much slower rate. We also found that the immortalization of normal human prostatic epithelial cells by SV40 T antigen led to a reduction in LRAT protein expression and esterification of [(3)H]retinol. Further transformation to tumorigenicity with the ras oncogene resulted in loss of detectable LRAT expression. Finally, we analyzed LRAT protein expression in tissue sections from six prostatectomy specimens by immunohistochemistry. LRAT protein was predominantly expressed in the basal cells of normal prostatic epithelium, whereas its expression was lost in prostate cancer. Collectively, these data implicate aberrant retinoid metabolism in the process of prostatic carcinogenesis. PMID- 11912137 TI - Significance of MAD2 expression to mitotic checkpoint control in ovarian cancer cells. AB - Chromosome instability is a commonly observed feature in ovarian carcinoma. Mitotic checkpoint controls are thought to be essential for accurate chromosomal segregation, and MAD2 is a key component of this checkpoint. In this study, we investigated the competence of the mitotic checkpoint and its relationship to the expression of MAD2 protein in seven ovarian cancer cell lines. We found that a significant number (43%, three of seven cell lines) of the tested ovarian cancer cells failed to arrest in the G(2)-M phase of the cell cycle in response to microtubule disruption. This loss of mitotic checkpoint control was associated with reduced expression of the MAD2 protein. To additionally understand the significance of the MAD2 to mitotic checkpoint control, we established an inducible expression system in which MAD2 was induced by the addition of ponasterone A. Notably, the induced expression of MAD2 in two checkpoint defective ovarian cancer cell lines led to the restoration of mitotic checkpoint response to spindle-disrupting agents. Taken together, our findings suggest that the steady-state amount of MAD2 inside cells may represent a molecular switch for mitotic checkpoint control. This provides a novel insight into the molecular basis of CIN in ovarian carcinoma and has implications for effective use of checkpoint-targeting drugs. PMID- 11912138 TI - Vascular endothelial growth factor-D expression is an independent prognostic marker for survival in colorectal carcinoma. AB - The angiogenic factor vascular endothelial growth factor-D (VEGF-D) isa ligand for VEGF receptor-3 (VEGFR-3/Flt-4) and receptor-2 (VEGFR-2/KDR)and is implicated in the development of lymphatic vessels and promotion of lymphatic metastases. We assessed the expression of VEGF-D and VEGFR-3 in relation to microvessel density (MVD) in colorectal carcinomas (CRC), adenomas, and adjacent normal tissue by immunohistochemistry on consecutive archival sections. VEGF-D was detected in malignant and benign epithelium and in some smooth muscle of the colorectum. High grade VEGF-D expression was observed frequently (74%) in CRC compared with adenomas (0%) and adjacent normal mucosa (22%). High-grade VEGF-D expression was not correlated with MVD, Dukes' stage (A to C), or tumor differentiation, but was associated with lymphatic involvement and patient survival. By multivariate analysis, VEGF-D expression was found to be an independent prognostic factor for both disease-free and overall survival. VEGFR-3 expression was detected in a subset of vessels, typically thin-walled and devoid of RBCs, in 89% of CRC cases examined. VEGFR-3-positive vessel densities increased progressively from normal mucosa to adenomas and carcinomas and were correlated with MVD, but not with Dukes' stage (A to C), tumor differentiation, or VEGF-D expression. VEGFR-3 expression was spatially associated with macrophage-rich inflammatory infiltrates, which were significantly more frequent among VEGFR-3-positive cases. We conclude that VEGF-D expression, but not that of its receptor VEGFR-3, is an independent prognostic indicator in CRC. VEGF-D expression may be associated with disease outcome through the promotion of lymphatic involvement/metastases. PMID- 11912139 TI - Cyclooxygenase-2 expression in human breast cancers and adjacent ductal carcinoma in situ. AB - Cyclooxygenase-2 (COX-2) is an inducible enzyme that converts arachidonic acid to prostaglandins. Overexpression of the COX-2 gene in mammary glands of transgenic mice was sufficient to induce tumorigenesis. We analyzed COX-2 expression in human breast cancers (and breast cancer cell lines) and adjacent ductal carcinoma in situ (DCIS) as well as its association with HER2/neu and clinicopathological variables. Archival primary breast carcinomas (n = 57), adjacent DCIS (n = 14) and DCIS alone (n = 2) were analyzed for COX-2 and HER2 expression by immunohistochemistry using specific monoclonal antibodies. An immunohistochemical scoring system was used. HER2 gene amplification had been analyzed previously by fluorescence in situ hybridization (n = 20). Histology of carcinomas included infiltrating ductal (n = 44), lobular (n = 2), and other (n = 7). Frozen breast cancers and adjacent normal tissue pairs (n = 9) were analyzed for COX-2 mRNA by reverse transcription-PCR. COX-2 and HER2 expression were also analyzed in human breast cancer cell lines (MCF-7, MCF-7/HER2, SK-BR-3, and MDA-MB-231) by immunoblotting. Cytoplasmic COX-2 expression was detected at an intermediate or high level in epithelial cells in 18 of 42 (43%) invasive breast cancers and in 10 of 16 (63%) cases of DCIS. Normal-appearing breast epithelia adjacent to cancer expressed COX-2 in 81% of cases and was generally focal and of similar or decreased intensity relative to adjacent neoplastic epithelia. COX-2 mRNA was detected in all samples analyzed by reverse transcription-PCR and was increased in eight of nine breast cancers relative to paired normal tissue. In archival tumors, no significant correlation was found between COX-2 and HER2 expression/amplification and clinicopathological variables. COX-2 expression was induced in MCF-7 cells stably transfected with HER2, in contrast to parental MCF 7 cells, and was detected in MDA-MB-231, but not SK-BR-3 cells. COX-2 is frequently overexpressed in invasive breast cancers and in adjacent DCIS and, thus, may be an early event in mammary tumorigenesis. Forced HER2 expression in MCF-7 cells was shown to up-regulate COX-2, although no association was found in human tumors. Our results suggest that nonsteroidal anti-inflammatory drugs and selective COX-2 inhibitors may be useful in the chemoprevention and therapy of human breast cancer. PMID- 11912140 TI - Endogenous von Hippel-Lindau tumor suppressor protein regulates catecholaminergic phenotype in PC12 cells. AB - Loss of von Hippel-Lindau (VHL) gene function leads to VHL disease, which is characterized by vascular tumors of the central nervous system, renal clear cell carcinomas, and pheochromocytomas. Pheochromocytomas express high levels of tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. PC12 cells that express VHL antisense RNA had 5-10-fold reduced levels of endogenous pVHL and 2-3-fold increased levels of TH protein and mRNA. Nuclear run-on analysis revealed an augmentation of TH gene transcription with enhanced efficiency of transcript elongation in the 3' region of the gene. Transient coexpression of the VHL antisense RNA with a TH promoter reporter construct increased TH promoter activity by 2-3-fold. A decrease in pVHL accumulation also resulted in an increase in TH mRNA accumulation and transcription of the TH gene during hypoxia. This is the first evidence that endogenous pVHL is a physiological regulator of the catecholaminergic phenotype. Thus, loss of pVHL function may be causative in pheochromocytoma-associated hypercatecholaminemia and arterial hypertension. PMID- 11912141 TI - Transcriptional regulation of mitotic genes by camptothecin-induced DNA damage: microarray analysis of dose- and time-dependent effects. AB - cDNA microarray technology can be used to establish associations between characteristic gene expression patterns and molecular responses to drug therapy. In this study, we used cDNA microarrays of 1694 cancer-related genes to monitor the gene expression consequences of the treatment of HCT116 colon cancer cells with the topoisomerase I inhibitor camptothecin (CPT). To obtain a more homogeneous cellular response, we synchronized the cells in S-phase using aphidicolin (APH) before CPT treatment. Brief incubation with 20 and 1000 nM CPT caused reversible and irreversible G(2) arrest, respectively, and the patterns of gene expression change (with reference to untreated controls) were strikingly different at the two concentrations. Thirty-three genes, mainly divided into three groups, showed characteristic changes in the first 20 h as a consequence of treatment. Northern blots performed for five of these genes (each under eight experimental conditions) were quite consistent with the microarray results (average correlation coefficient, 0.86). Several p53-activated stress response genes were up-regulated after treatment with 1000 nM CPT or prolonged exposure to APH, but it seemed that the up-regulation did not directly cause cell cycle arrest because the up-regulation induced by prolonged treatment with APH did not prevent cell cycle progression after removal of APH. In contrast, cell cycle dependent up-regulation of a group of mitosis-related genes was delayed or blocked after CPT treatments. The interrupted up-regulation of this group of genes was directly associated with G(2) arrest. In addition, we observed down regulation of gene expression in cells that were recovering from cell cycle delay. The observations reported here suggest a fundamental difference at the gene expression level between the molecular mechanism of reversible G(2) delay that follows mild DNA damage and the mechanism of permanent G(2) arrest that follows more extensive DNA damage. PMID- 11912142 TI - Oncolytic reovirus against ovarian and colon cancer. AB - Reovirus selectively replicates in and destroys cancer cells with an activated Ras signaling pathway. In this study, we evaluated the feasibility of using reovirus (serotype 3, strain Dearing) as an antihuman colon and ovarian cancer agent. In in vitro studies, reovirus infection in human colon and ovarian cell lines was assessed by cytopathic effect as detected by light microscopy, [(35)S]Methionine labeling of infected cells for viral protein synthesis and progeny virus production by plaque assay. We observed that reovirus efficiently infected all five human colon cancer cell lines (Caco-2, DLD-1, HCT-116, HT-29, and SW48) and four human ovarian cancer cell lines (MDAH2774, PA-1, SKOV3, and SW626) which were tested, but not a normal colon cell line (CCD-18Co) or a normal ovarian cell line (NOV-31). We also observed that the Ras activity in the human colon and ovarian cancer cell lines was elevated compared with that in normal colon and ovarian cell lines. In animal models, intraneoplastic as well as i.v. inoculation of reovirus resulted in significant regression of established s.c. human colon and ovarian tumors implanted at the hind flank. Histological studies revealed that reovirus infection in vivo was restricted to tumor cells, whereas the surrounding normal tissue remained uninfected. Additionally, in an i.p. human ovarian cancer xenograft model, inhibition of ascites tumor formation and the survival of animals treated with live reovirus was significantly greater than of control mice treated with UV-inactivated reovirus. Reovirus infection in ex vivo primary human ovarian tumor surgical samples was also confirmed, further demonstrating the potential of reovirus therapy. These results suggest that reovirus holds promise as a novel agent for human colon and ovarian cancer therapy. PMID- 11912143 TI - Simultaneous inhibition of the receptor kinase activity of vascular endothelial, fibroblast, and platelet-derived growth factors suppresses tumor growth and enhances tumor radiation response. AB - We have investigated the effect of simultaneous inhibition of multiple angiogenic growth factor signaling pathways on tumor growth, tumor blood perfusion, and radiation-induced tumor-growth delay using SU6668, an inhibitor of the receptor tyrosine kinase activity of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and platelet-derived growth factor (PDGF). The SCK mammary carcinoma, FSaII fibrosarcoma, and CFPAC human pancreatic carcinoma were grown s.c. in the hind leg of A/J mice, C3H mice, and Balb/cAnNCrl-nuBr nude mice, respectively. Daily i.p. injection of 100 mg/kg of SU6668 markedly suppressed the growth of these three tumor types. SU6668 also markedly prolonged the survival time of host mice bearing SCK tumors, which appeared to be caused by a reduction of metastatic tumor growth in the lung. There was little or no change in normal tissue blood perfusion, whereas in SCK tumors the perfusion decreased by 50% at 1 h after a single i.p. injection of SU6668, slightly recovered at 4 h, and completely recovered by 8 h. Interestingly, the tumor blood flow was significantly increased above the baseline level 24 h after SU6668 injection. After extended daily i.p. injections of SU6668, the tumor blood flow in all of the three tumor types studied was markedly decreased compared with control. The observed effects of this drug on tumor blood perfusion may partially explain the effectiveness of the compound in suppressing tumor growth and extending survival of tumor-bearing mice. We also observed that daily SU6668 administration and a single dose of 15 Gy of X-irradiation was significantly more effective than either treatment alone in suppressing tumor growth. Our results suggest that SU6668 increased the radiosensitivity of tumor blood vessels. We conclude that SU6668 is a potent therapeutic agent potentially useful to suppress tumor growth and enhance the response of tumors to radiotherapy. PMID- 11912144 TI - Selective sensitization of transformed cells to flavopiridol-induced apoptosis following recruitment to S-phase. AB - Flavopiridol is a potent inhibitor of cyclin-dependent kinases (cdks). In a large proportion of solid tumor cell lines, the initial response to flavopiridol is cell cycle arrest. NCI-H661 non-small cell lung cancer cells are representative of a subset of more sensitive cell lines in which apoptosis is observed during the first 24 h of drug exposure. Analysis of the apoptotic population indicates that cells in S-phase are preferentially dying. In addition, cells are sensitized to flavopiridol following recruitment to S-phase, whether accomplished by synchronization or by treatment with noncytotoxic concentrations of chemotherapy agents that impose an S-phase delay. Combinations of gemcitabine or cisplatin, followed by flavopiridol at concentrations that correlate with cdk inhibition, produce sequence-dependent cytotoxic synergy. A survey of paired cell lines, including WI38 diploid fibroblasts or normal human bronchial epithelial cells, along with their SV40-transformed counterparts, demonstrates that treatment with flavopiridol during S-phase is selectively cytotoxic to transformed cells. These data suggest that treatment during S-phase may maximize responses to flavopiridol and that the administration of flavopiridol after chemotherapy agents that cause S-phase accumulation may be an efficacious antitumor strategy. PMID- 11912145 TI - The new sulindac derivative IND 12 reverses Ras-induced cell transformation. AB - The nonsteroidal anti-inflammatory drug Sulindac has chemopreventive and antitumorigenic properties. Its metabolites induce apoptosis and inhibit signaling pathways critical for malignant transformation, including the Ras pathway. Here we show that the new Sulindac derivative IND 12 reverses the phenotype of Ras-transformed MDCK-f3 cells and restores an untransformed epithelioid morphology characterized by growth in monolayers with regular cell cell adhesions. Moreover, IND 12 treatment induces the expression at membranes of the cell adhesion protein E-cadherin and increases the level of the E-cadherin bound beta-catenin. As a consequence, IND 12-treated MDCK-f3 cells lose their invasion capacity and regain the ability to aggregate. In the presence of IND 12, MDCK-f3 cells show regenerated expression and activity ratios of the small GTPases Rac and Rho normally found in untransformed MDCK cells. Strikingly, IND 12 treatment decreases the levels of phosphorylated mitogen-activated protein kinases, which are downstream substrates of the Ras-regulated Raf/mitogen activated protein kinase pathway, and the level of Ras-induced activation of gene expression. Our findings identify a novel drug with high potential in cancer therapy by targeting Ras-induced cell transformation. PMID- 11912146 TI - Three new prodrugs for suicide gene therapy using carboxypeptidase G2 elicit bystander efficacy in two xenograft models. AB - Three new prodrugs, [prodrug 1: 4-[bis(2-iodoethyl)amino]-phenyloxycarbonyl-L glutamic acid; prodrug 2: 3-fluoro-4-[bis(2-chlorethyl)amino]benzoyl-L-glutamic acid; and prodrug 3: 3,5-difluoro-4-[bis(2-iodoethyl)amino]benzoyl-L-glutamic acid] have been assessed for use with a mutant of carboxypeptidase G2 (CPG2, glutamate carboxypeptidase, EC 3.4.17.11,) engineered to be tethered to the outer tumor cell surface (stCPG2(Q)3) as the activating enzyme in suicide gene therapy systems. All three of the prodrugs produce much greater cytotoxicity differentials between stCPG2(Q)3- and control beta-galactosidase (beta-gal) expressing breast carcinoma MDA MB 361 and colon carcinoma WiDr cells (70- to 450 fold) than was previously observed (19- to 27-fold) with 4-[(2-chloroethyl)(2 mesyloxyethyl)amino]benzoyl-L-glutamic acid (CMDA). Prodrug 1 is the most effective antitumor agent in xenografts in mice inoculated with 100% stCPG2(Q)3 expressing MDA MB 361 cells, whereas prodrugs 2 and 3 are most effective when the percentage of stCPG2(Q)3-expressing cells is 50% or 10%. In nude mice bearing xenografts arising from inocula of 100% stCPG2(Q)3-expressing WiDr cells, prodrug 2 is the most effective antitumor agent. All three of the prodrugs produced histological evidence of substantial bystander cell killing in WiDr xenografts in which only 10% or 50% of the cells inoculated were expressing stCPG2(Q)3. We conclude that all three of the prodrugs are more effective therapeutically with stCPG2(Q)3 than is the previously described prodrug CMDA and, also, that the optimal choice of prodrug varies among different tumor types and that prodrugs, optimized for their bystander effect, are effective when only low percentages of cells in a tumor express CPG2. PMID- 11912147 TI - A diphtheria toxin-interleukin 3 fusion protein is cytotoxic to primitive acute myeloid leukemia progenitors but spares normal progenitors. AB - The relative cytotoxicity of a diphtheria toxin (DT) human interleukin 3(IL3) fusion protein (DT(388)IL3) was tested against primitive normal (n = 3)and acute myeloid leukemia (AML) progenitors (n = 7). After 24-h culture with 50 ng/ml DT(388)IL3, the mean percentages of kill of AML colony-forming cells (CFCs), long term culture-initiating cells (LTC-ICs), and suspension culture-ICs (SC-ICs) were 82% (range, 47-100), 56% (range, 28-91), and 74% (range, 43-87), respectively, with most surviving progenitors being cytogenetically normal. Engraftment of DT(388)IL-3-treated AML cells in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice followed for 16 weeks was eradicated for two of these samples. In contrast, with normal bone marrow, mean percentages of CFC kill of 49 and 64% were seen with 50 or 250 ng/ml DT(388)IL3, respectively, whereas no significant kills were observed in the LTC-IC and SC-IC assays. The NOD/SCID mouse repopulating cell (RC) frequency in normal BM cells was also not reduced by DT(388)IL3 treatment. In subsequent experiments, NOD/SCID mice that received AML blasts i.v. followed in 24 h by 0.045 microg/g DT(388)IL3 daily i.p. x 5 showed mean percentages of reduction in AML engraftment of 83% (range, 14-100) and 57% (range, 0-98) after 4 and 12 weeks, respectively (n = 6). No evidence of leukemia was detected with two of six AML samples 12 weeks after one 5-day course of DT(388)IL3. Repeating the DT(388)IL3 treatment every 4 weeks enhanced its effectiveness against two additional samples. Thus, DT(388)IL3 kills primitive leukemic progenitors from a proportion of AML patients but shows no significant toxicity against equivalent normal cells. PMID- 11912148 TI - Development of a recombinant HSP110-HER-2/neu vaccine using the chaperoning properties of HSP110. AB - Several studies have shown that when purified from a tumor, certain heat shock proteins (HSPs) can function as effective vaccines against the same tumor by virtue of their ability to bind tumor-specific peptides. However, only a small fraction of the associated peptides would be expected to be immunogenic, in addition to which, the clinical application of this vaccine requires the availability of a surgical specimen of sufficient quantity for purification of the HSP. The present study describes a new approach for the development of natural HSP vaccines that do not have these limitations. This approach uses a recombinant HSP that is noncovalently bound to a recombinant tumor protein antigen by heat shock. HSP110 has been selected for this purpose, because it has been shown to be a highly efficient molecular chaperone in binding to large protein substrates. We show that a "natural chaperone complex" between HSP110 and the intracellular domain (ICD) of human epidermal growth factor receptor 2 protein (HER-2)/neu is formed by heat shock. This HSP110-ICD vaccine elicited both CD8(+) and CD4(+) T-cell responses against ICD as determined by an antigen specific IFN-gamma production in an enzyme-linked immunospot assay (ELISPOT). In vivo depletion studies revealed that the CD8(+) T-cell response was independent of CD4(+) T-cell help. The HSP110-ICD complex also significantly enhanced ICD specific antibody responses relative to that seen with ICD alone. No CD8(+) T cell or antibody response was detected against HSP110. The use of recombinant HSP110 to form natural chaperone complexes with large protein antigens represents a new and powerful approach for the design of protein-targeted cancer vaccines. PMID- 11912149 TI - Circulating Tumor-reactive CD8(+) T cells in melanoma patients contain a CD45RA(+)CCR7(-) effector subset exerting ex vivo tumor-specific cytolytic activity. AB - To defend the host from malignancies, the immune system can spontaneously raise CD8(+) T-cell responses against tumor antigens. Investigating the functional state of tumor-reactive cytolytic T cells in cancer patients is a key step for understanding the role of these cells in tumor immunosurveillance and for evaluating the potential of immunotherapeutic approaches of vaccination against cancer. In this study we identified a subset of circulating tumor-reactive CD8(+) T lymphocytes, which specifically secreted IFN-gamma after exposition to autologous tumor cell lines in stage IV metastatic melanoma patients. Additional phenotypic characterization using multicolor flow cytometry revealed that a significant fraction of these cells were CD45RA(+)CCR7(-), a phenotype that has been proposed recently to characterize cytolytic effectors potentially able to home into inflamed tissues. In the case of an HLA-A2-expressing patient, the antigen specificity of this population was identified by using HLA-A2/peptide multimers incorporating a tyrosinase-derived peptide. Consistently with their phenotypic characteristics, A2/tyrosinase peptide multimer(+) CD8(+) T cells, isolated by cell sorting, were directly lytic ex vivo and able to specifically recognize tyrosinase-expressing tumor cells. Overall, these results provide the first evidence that a proportion of melanoma patients have circulating tumor reactive T cells, which are lytic effectors cells. PMID- 11912150 TI - Tumor-infiltrating B lymphocytes as a potential source of identifying tumor antigen in human lung cancer. AB - Functional studies using freshly isolated tumor-infiltrating B lymphocytes(TIB) are difficult to perform and interpret. Here we document a novel function of TIB using fresh human lung cancer tissues engrafted in SCID mice; they are at activated state and produce tumor-specific antibodies in tumor microenvironment: (a) TIB engrafted in SCID mice produced human IgG; (b) IgG derived from TIB highly bound intracellular and membrane-bound antigens of autologous cancer cells; and (c) less recognition of autoantigens expressed on normal lymphocytes by IgG derived from TIB compared with IgG from the serum of the patient. On the basis of the novel findings presented in this study, we modified the original serological analysis of antigens by recombinant cDNA expression cloning design in a patient with lung cancer who expressed unusually favorable clinical evolution and analyzed humoral immunity against identified mutated p53 antigen. This study provides the first demonstration that antibodies derived from TIB recognize tumor antigens by serological analysis of antigens by recombinant cDNA expression cloning methodology and circulating anti-p53 antibodies in sera derived from TIB in tumor microenvironment. Our approach using TIB may allow the identification of key antigens in the humoral cancer-related immune system. PMID- 11912152 TI - Novel regions of allelic imbalance identified by genome-wide analysis of neuroblastoma. AB - Several nonrandom chromosomal abnormalities have been associated with neuroblastoma (NB). However, the relationship of each genetic event to the clinical course of disease is not firmly established. We have performed a genome wide allelic scan of NB to identify regions with frequent allelic imbalance (AI) and correlate the allelotype with clinical features of disease. Nineteen tumors from patients across the spectrum of NB were used. Genome-wide allelotype was performed using 169 fluorescently labeled microsatellite markers from the Weber 9a human screening set (Research Genetics, Huntsville, AL) and 48 independent markers for high-density analysis of selected regions. Eleven chromosomal regions had AI in >25% of tumors including loci known previously to be frequently altered such as 1p36 (10 of 19; 52%), 2p (9 of 19; 47%), 17q (8 of 19; 42%), 11q23 (8 of 19; 42%), 14q32 (7 of 19; 37%), 19q (6 of 19; 31%), 7q (6 of 19; 31%), 9p21 (5 of 19; 26%), and three novel regions of frequent AI at 10p11-p15 (7 of 19; 40%), 12q24.1 (5 of 19; 26%), and 8qcen-q24 (5 of 19; 26%). AI of four regions (8q, 10p, 19q, and 12q) allowed the distinction of two genetic groups that matched clinical significant subgroups of NB. AI at 12q24 and 19q13 was found exclusively in high-risk local-regional tumors, whereas AI at 10p11 and 8q appeared to be specific for stage 4 tumors with MCYN amplification. Spontaneously remitting or quiescent tumors were intact at all of the regions described above. PMID- 11912151 TI - Vaccination with DNA encoding a single-chain TCR fusion protein induces anticlonotypic immunity and protects against T-cell lymphoma. AB - The clonotypic T-cell antigen receptor (TCR) provides unique Valpha and Vbeta sequences with potential as idiotypic targets for immunoregulation. For T-cell malignancies, vaccination with the TCR could induce therapeutic anti-idiotypic responses. To facilitate this approach, we have developed DNA vaccines that include the genes encoding TCR sequences from a T-cell lymphoma (TCL). To combine requirements for stable folding with a simple minimized single-chain construction, we used a three-domain ValphaVbetaCbeta sequence. To promote anti TCR immunity, we fused a pathogen-derived sequence from tetanus toxin to the 3' end of the single-chain TCR. The fusion gene vaccine induced anti-idiotypic antibodies and generated protection against the TCL. The critical requirement for the conformational integrity of the delivered TCR antigen was highlighted by the observation that DNA fusion vaccines containing either ValphaVbeta or VbetaCbeta sequences failed to generate antibodies reactive with the native TCR or provide protection. This is the first report of a DNA vaccine able to induce anti idiotypic immunity against TCL, and it presents a simple strategy for selectively eliminating T-cell clones in vivo. PMID- 11912153 TI - In lymphatic cells par-4 sensitizes to apoptosis by down-regulating bcl-2 and promoting disruption of mitochondrial membrane potential and caspase activation. AB - Inhibition of apoptosis is a hallmark of malignancies of the hematopoetic system. Previous studies in nonhematopoetic cells demonstrated that the prostate apoptosis-response-gene-4 (Par-4) is up-regulated in cells undergoing programmed cell death and that Par-4 exerts its proapoptotic effect by down-regulating Bcl 2. After showing the aberrant expressional pattern of Par-4 in neoplastic lymphocytes as well as demonstrating inverse expressional patterns of Par-4 and Bcl-2 in malignant cells of patients suffering from acute lymphocytic leukemia, we assessed the functional consequences of Par-4 overexpression during apoptosis in Jurkat T lymphocytes. We show that in lymphatic cells Par-4 overexpression decreases the level of Bcl-2, whereas Bax, the proapoptotic counterpart of Bcl-2, retains unaltered levels. Moreover, Par-4 overexpression is accompanied by cleavage of poly(ADP-ribose) polymerase (PARP). Despite these effects, overexpression of Par-4 alone is not sufficient to induce apoptosis but markedly increases the rate of apoptosis on treatment with different chemotherapeutic agents. On chemotherapeutic treatment Par-4 overexpression enhances disruption of mitochondrial membrane potential, PARP-cleaving activity, as well as activation of caspase-3. The hypothesis of caspase-dependency of Par-4-promoted apoptosis is additionally supported by demonstrating complete abrogation of programmed cell death after pretreatment with a broad spectrum caspase-inhibitor. On inhibition of caspase-3 overexpression of Par-4 enables lymphatic cells to alternatively activate caspases-9, -6, and -7 by diminishing the influence of the inhibitors of apoptosis proteins (IAPs) cIAP1 and XIAP. Our study is the first to identify Par 4 as a proapoptotic protein in lymphatic cells, outlining a model of action evaluating the role of Bcl-2/Bax, as well as demonstrating the impact of Par-4 expression on PARP cleavage, disruption of mitochondrial membrane potential, caspase activation, and interactions with inhibitors of apoptosis proteins. PMID- 11912154 TI - Adenovirus-mediated E2F-1 gene transfer sensitizes melanoma cells to apoptosis induced by topoisomerase II inhibitors. AB - Melanoma has proven to be resistant to conventional chemotherapy; however,the mechanism of chemoresistance is still unclear. Recent reports show that the transcription factor, E2F-1, may play a role in mediating cytotoxicity of certain chemotherapeutic agents. We have shown in a previous study that adenovirus mediated overexpression of E2F-1 can efficiently induce apoptosis in melanoma cells. In the present study, the effect of E2F-1 expression on drug sensitivity of melanoma cells was evaluated. Two human melanoma cell lines, SK-MEL-28 and SK MEL-2, were treated with drugs (etoposide, Adriamycin, roscovitine, cisplatin, 5 fluorouracil, or cycloheximide), alone or in combination with adenoviral vectors expressing beta-galactosidase (Ad-LacZ) or E2F-1 (Ad-E2F-1) at a multiplicity of infection of 1 in vitro. E2F-1 expression was confirmed by Western blot analysis. Sublethal concentrations of each drug alone or infection with Ad-E2F-1 alone produced <5% apoptosis by 3 days posttreatment. Conversely, cotreatment with Ad E2F-1 and low concentrations of etoposide or Adriamycin markedly sensitized melanoma cells to apoptotic cell death. A slight enhancement of the cytotoxicity of roscovitine was demonstrated in combination with E2F-1 overexpression, but not to cisplatin, 5-fluorouracil, or cycloheximide. Ad-LacZ infection showed no obvious effects on drug sensitivity. Overexpression of p21 can block apoptosis induced by the combination chemogene therapy of Ad-E2F-1 and topoisomerase II poisons and does not require its proliferating cell nuclear antigen-binding ability. The protein synthesis inhibitor cycloheximide also has a cytotoxicity protective effect against topoisomerase II inhibitor/E2F-1-induced apoptosis and suggests that new protein synthesis is required for this process. Topoisomerase II inhibitors also cooperated with Ad-E2F-1 to enhance antitumor activity in an in vivo model using xenografts in nude mice. When combined with Adriamycin or etoposide, E2F-1 adenovirus therapy resulted in an 87% or 91% decrease in tumor size, respectively, compared with controls (P < 0.002). Our results show that adenovirus-mediated E2F-1 gene transfer can sensitize melanoma cells to some chemotherapeutic agents, particularly topoisomerase II poisons, in vitro and in vivo. These results suggest a new chemosensitization strategy for melanoma gene therapy. PMID- 11912155 TI - Joint effect of HSD3B1 and HSD3B2 genes is associated with hereditary and sporadic prostate cancer susceptibility. AB - 3beta-hydroxysteroid dehydrogenases (HSD3Bs), encoded by the HSD3B gene family at 1p13, have long been hypothesized to have a major role in prostate cancer susceptibility. The recent reports of a prostate cancer linkage at 1p13 provided additional evidence that HSD3B genes may be prostate cancer susceptibility genes. To evaluate the possible role of HSD3B genes in prostate cancer, we screened a panel of DNA samples collected from 96 men with or without prostate cancer for sequence variants in the putative promoter region, exons, exon-intron junctions, and 3'-untranslated region of HSD3B1 and HSD3B2 genes by direct sequencing. Eleven single nucleotide polymorphisms (SNPs) were identified, four of which, including a missense change (B1-N367T), were informative. These four SNPs were further genotyped in a total of 159 hereditary prostate cancer probands, 245 sporadic prostate cancer cases, and 222 unaffected controls. Although a weak association between prostate cancer risk and a missense SNP (B1-N367T) was found, stronger evidence for association was found when the joint effect of the two genes was considered. Men with the variant genotypes at either B1-N367T or B2 c7519g had a significantly higher risk to develop prostate cancer, especially the hereditary type of prostate cancer. Most importantly, the subset of hereditary prostate cancer probands, whose families provided evidence for linkage at 1p13, predominantly contributed to the observed association. Additional studies are warranted to confirm these findings. PMID- 11912156 TI - DMBT1 polymorphisms: relationship to malignant glioma tumorigenesis. AB - The deleted in malignant brain tumors 1 (DMBT1) gene on 10q25-26 is a candidate tumor suppressor gene in malignant gliomas, but its role is controversial, e.g., some DMBT1 homozygous deletions reflect unmasking of constitutional deletion polymorphisms by 10q loss. To clarify the role of DMBT1 in gliomagenesis, we investigated three reported deletion hot spots. Homozygous deletions at DMBT1 repeat 2-4 to 2-7 were found in 10 of 73 gliomas with 10q loss, but all 10 deletions reflected unmasking of constitutional hemizygous deletions. Alleles bearing deletion 2-4/2-7 were not selected significantly for by 10q loss, with retention of only 10 of 16 deleted alleles. No homozygous deletion was detected at locus 74k in the 5' upstream region of DMBT1, and four tightly linked polymorphisms were found around this region; chromosome 10q loss randomly affected alleles with or without the variant sequences around locus 74k. Moreover, no significant selection pressure was detected for the haplotype with both deletion 2-4/2-7 and 5' polymorphisms. There was no segregation of deletion 2-4/2-7 in glioma patients compared with unrelated individuals from reference families but a suggestion of a difference in the distribution of the 5' polymorphisms between the reference individuals and glioma patients. Constitutional polymorphisms at DMBT1 repeat 2-9/2-10 appeared common in patients with both benign brain tumors and gliomas. A homozygote for both the 2-4/2-7 deletion and the 5' polymorphisms had a glioma arise at a typical age and without an apparent family cancer predisposition. These data suggest that DMBT1 polymorphisms are not likely primary targets of 10q loss in malignant gliomas and do not support a major role for DMBT1 in gliomagenesis. PMID- 11912157 TI - Chfr regulates a mitotic stress pathway through its RING-finger domain with ubiquitin ligase activity. AB - Resistance to chemotherapy targeting microtubules could be partially because of the delay in chromosome condensation and segregation during mitosis. The Chfr pathway has been defined recently, and its activation causes a delay in chromosome condensation in response to mitotic stress. Because Chfr contains a RING-finger domain, we tested whether Chfr inhibits chromosome condensation through an ubiquitin (ubiquitin)-dependent pathway. In the presence of purified E1, Ubc4, or Ubc5, and ubiquitin, Chfr catalyzes its own ubiquitination in vitro, an activity requiring the RING domain. In vivo, overexpressed Chfr but not a RING domain mutant is spontaneously ubiquitinated. Our studies with DLD1 cells stably expressing wild-type Chfr and Chfr lacking the RING domain indicated that the RING-finger deletion mutant was defective in inhibiting chromosome condensation after Taxol treatment. In addition, Chfr expression increases the survival rate after Taxol treatment, an activity requiring the RING domain. Preliminary studies indicate that Chfr expression is cell cycle regulated and is dependent on its ubiquitin ligase activity. It is very likely that the Chfr-mediated ubiquitin dependent pathway is a critical component of the response to mitotic stress. PMID- 11912158 TI - Expression of the neurotrophin receptor TrkA down-regulates expression and function of angiogenic stimulators in SH-SY5Y neuroblastoma cells. AB - Angiogenesis is essential for tumor growth and metastasis and depends on the production of angiogenic factors. Mechanisms regulating the expression of angiogenic factors in tumor cells are largely unknown. High expression of the neurotrophin receptor TrkA in neuroblastomas (NBs) is associated with a favorable prognosis, whereas TrkB is mainly expressed on aggressive, MYCN-amplified NBs. To investigate the biological effects of TrkA and TrkB expression on angiogenesis in NB, we examined the expression of angiogenic factors in the human NB cell line SY5Y and its TrkA and TrkB transfectants. In comparison with parental SY5Y cells, mRNA and protein levels of the examined angiogenic factors were significantly reduced in SY5Y-TrkA cells, whereas SY5Y-TrkB cells did not demonstrate a significant change. Conditioned medium of TrkB transfectants and parental SY5Y cells induced endothelial cell proliferation and migration, but this effect was completely absent in SY5Y-TrkA cells. TrkA expression also resulted in severely impaired tumorigenicity in a mouse xenograft model and was associated with reduced angiogenic factor expression and vascularization of tumors, as determined by immunohistochemistry and an in vivo Matrigel assay. TrkA expression inhibits angiogenesis and tumor growth in SY5Y NB cells by down-regulation of angiogenic factors, whereas expression of TrkB does not down-regulate the production of these angiogenic factors. The biologically different behavior of TrkA- and TrkB expressing NBs may be explained in part by their effects on angiogenesis. PMID- 11912159 TI - Nuclear factor kappaB dependency of platelet-activating factor-induced angiogenesis. AB - This study investigated the mechanisms of platelet-activating factor (PAF) induced angiogenesis in a mouse model of Matrigel implantation. PAF induced a dose- and time-dependent angiogenic response. Inhibitors of nuclear factor (NF) kappaB expression or action, including antisense oligonucleotides to the p65 subunit of NFkappaB (p65 antisense) and antioxidants such as alpha-tocopherol and N-acetyl-L-cysteine, significantly reduced PAF-induced angiogenesis. In human umbilical vein endothelial cells, PAF-induced mRNA expression and protein synthesis of various NFkappaB-dependent angiogenic factors, such as tumor necrosis factor-alpha, interleukin-1alpha, basic fibroblast growth factor, and vascular endothelial growth factor (VEGF). The PAF-induced expression of the above mentioned factors was inhibited by p65 antisense or antioxidants. A significant inhibition of the angiogenic effect of PAF was achieved by anti-VEGF antibodies or soluble VEGF receptors such as KDR and flt-1 but not by antibodies against tumor necrosis factor-alpha, interleukin-1alpha, or basic fibroblast growth factor. These data indicate that PAF enhances angiogenesis through inducing NFkappaB activation, which in turn promotes the production of angiogenic factors such as VEGF. PMID- 11912160 TI - Atypical protein kinase C zeta as a target for chemosensitization of tumor cells. AB - Exposure of tumor cells to cytotoxic agents simultaneously activates a variety of intracellular signaling pathways. Some of these pathways involve enzymes from the protein kinase C (PKC) family of serine/threonine kinases. This family includes isoenzymes that negatively influence cell death, whereas other demonstrate an opposite effect. The present study analyzes the role of the zeta atypical PKC isoform in tumor cell response to cytotoxic agents. Using a histone H1 phosphorylation assay, we showed that both tumor necrosis factor alpha and etoposide activate PKCzeta in U937 human leukemic cells. Stable transfection of a kinase-dead, dominant-negative PKCzeta mutant in U937 cells decreases Bcl-2 expression while increasing the expression of Bax and several procaspases. This transfection also prevents etoposide-induced nuclear factor-kappaB nuclear translocation and accumulation of X-linked inhibitor of apoptosis protein. PKCzeta inhibition accelerates the occurrence of apoptosis in leukemic cells exposed to etoposide and tumor necrosis factor alpha. This sensitization was confirmed in vitro by use of a clonogenic assay. In addition, PKCzeta inhibition sensitized tumor cells grown in nude mice to etoposide. These results indicate that PKCzeta isoform is a protective signals that is activated in tumor cells exposed to a cytotoxic agent. This inducible resistance factor thus appears an attractive target for chemosensitization of tumor cells. PMID- 11912161 TI - Coamplification of DAD-R, SOX5, and EKI1 in human testicular seminomas, with specific overexpression of DAD-R, correlates with reduced levels of apoptosis and earlier clinical manifestation. AB - Seminomas and nonseminomas represent the invasive stages of testicular (TGCTs) of adolescents and adults. Although TGCTs are characterized by extra copies of the short arm of chromosome 12, the genetic basis for gain of 12p in the pathogenesis of this cancer is not yet understood. We have demonstrated that gain of 12p is related to invasive growth and that amplification of specific 12p sequences, i.e., 12p11.2-p12.1, correlates with reduced apoptosis in the tumors. Here we show that three known genes map within the newly determined shortest region of overlap of amplification (SROA): DAD-R, SOX5, and EKI1. Whereas EKI1 maps close to the telomeric region of the SROA, DAD-R is the first gene at the centromeric region within the 12p amplicon. Although all three genes are amplified to the same level within the SROA, expression of DAD-R is significantly up-regulated in seminomas with the restricted 12p amplification compared with seminomas without this amplicon. DAD-R is also highly expressed in nonseminomas of various histologies and derived cell lines, both lacking such amplification. This finding is of particular interest because seminomas with the restricted 12p amplification and nonseminomas are manifested clinically in the third decade of life and show a low degree of apoptosis. In contrast, seminomas lacking a restricted 12p amplification, showing significantly lower levels of DAD-R with pronounced apoptosis, manifest clinically in the fourth decade of life. A low level of DAD-R expression is also observed in normal testicular parenchyma and in parenchyma containing the precursor cells of this cancer, i.e., carcinoma in situ. Therefore, elevated DAD-R expression in seminomas and nonseminomas correlates with invasive growth and a reduced level of apoptosis associated with an earlier clinical presentation. These data implicate DAD-R as a candidate gene responsible in part for the pathological effects resulting from gain of 12p sequences in TGCTs. In addition, our results also imply differences in expression regulation of DAD-R between seminomas and nonseminomas. PMID- 11912162 TI - Use of the stromal cell-derived factor-1/CXCR4 pathway in prostate cancer metastasis to bone. AB - Neoplasms have a striking tendency to metastasize or "home" to bone. Hematopoietic cells also home to bone during embryonic development, where evidence points to the chemokine stromal cell-derived factor-1 (SDF-1 or CXCL12; expressed by osteoblasts and endothelial cells) and its receptor (CXCR4) as key elements in these processes. We hypothesized that metastatic prostate carcinomas also use the SDF-1/CXCR4 pathway to localize to the bone. To test this, levels of CXCR4 expression were determined for several human prostate cancer cell lines by reverse transcription-PCR and Western blotting. Positive results were obtained for cell lines derived from malignancies that had spread to bone and marrow. Prostate cancer cells were also observed migrating across bone marrow endothelial cell monolayers in response to SDF-1. In in vitro adhesion assays, pretreatment of the prostate cancer cells with SDF-1 significantly increased their adhesion to osteosarcomas and endothelial cell lines in a dose-dependent manner. Invasion of the cancer cell lines through basement membranes was also supported by SDF-1 and inhibited by antibody to CXCR4. Collectively, these results suggest that prostate cancers and perhaps other neoplasms may use the SDF-1/CXCR4 pathway to spread to bone. PMID- 11912163 TI - Vascular endothelial growth factor isoform expression as a determinant of blood vessel patterning in human melanoma xenografts. AB - Vascular endothelial growth factor (VEGF) occurs in at least five different isoforms because of alternative splicing of the gene. To investigate the roles of different VEGF isoforms in tumor blood vessel formation and tumorigenicity, the three major isoforms (VEGF(121), VEGF(165), and VEGF(189)) were overexpressed in an early-stage human melanoma cell line (WM1341B), which is VEGF-negative and nontumorigenic in immunodeficient mice. Although overexpression of VEGF(121) and VEGF(165) resulted in aggressive tumor growth, WM1341B cells transfected with VEGF(189) remained nontumorigenic and dormant on injection. Although tumor growth rate depended on the level and not the isoform of VEGF expressed, striking isoform-specific differences in vascular patterning were associated with VEGF(121)- versus VEGF(165)-dependent tumorigenic conversion of human melanoma. Thus, tumors overexpressing VEGF(165) generated dense, highly heterogeneous vessel networks that were distinctly different from those of tumors expressing VEGF(121) (poorly vascularized and necrotic). Paradoxically, although VEGF(165) expression appears to result in the most effective tumor perfusion, it is the expression of VEGF(121) that is observed during human malignant melanoma progression. Indeed, unbiased selection of spontaneously tumorigenic variants of WM1341B (by coinjection with Matrigel) led to predominant expression of the VEGF(121) isoform. The vascular patterning in these tumors (1341-P3N1, 1341-P3N2) resembled that of the VEGF(121)-transfected WM1341B tumors. These results suggest that, for reasons that remain to be elucidated, a "minimal" program of tumor vascularization may be favored during melanoma progression. PMID- 11912164 TI - Hypoxia promotes lymph node metastasis in human melanoma xenografts by up regulating the urokinase-type plasminogen activator receptor. AB - Clinical studies have shown that metastatic spread is associated with hypoxia in the primary tumor. The mechanism behind this association has not been identified and, in fact, it has not been established whether hypoxia induces metastasis or whether the most metastatic cell phenotypes develop the most hypoxic tumors. The present study demonstrates that hypoxia promotes spontaneous lymph node metastasis in R-18 human melanoma xenografts by up-regulating the urokinase-type plasminogen activator receptor (uPAR). Pimonidazole was used as a hypoxia marker, and hypoxia and uPAR expression were detected by immunohistochemistry. R-18 cells were capable of up-regulating uPAR under hypoxic conditions in vitro, as revealed by Western and Northern blot analyses, and uPAR-positive regions showed a high degree of colocalization with hypoxic regions in R-18 tumors. There was a strong correlation between uPAR-positive fraction and hypoxic fraction in individual tumors (P < 0.00001). Incidence of metastases, hypoxic fraction, and uPAR positive fraction increased with the size of the primary tumor with similar kinetics. Metastatic tumors showed approximately 1.5-fold higher hypoxic fraction (P = 0.00004) and approximately 1.4-fold higher uPAR-positive fraction (P = 0.0003) than nonmetastatic tumors of the same size. Moreover, treatment with neutralizing antibody against uPAR prevented metastasis almost completely. Only 1 of 30 treated mice developed metastases, whereas 14 of 30 control mice were metastasis positive, suggesting that functional uPAR is a prerequisite for lymph node metastasis in R-18 tumors. The study reported here suggests that metastatic spread may be promoted by hypoxia in the primary tumor and identifies the plasminogen activation system as an important target for the treatment of malignant melanoma. PMID- 11912165 TI - VEGF(121), VEGF(165) overexpression enhances tumorigenicity in U251 MG but not in NG-1 glioma cells. AB - Vascular endothelial growth factor (VEGF) is a multifunctional cytokine with important roles in angiogenesis. VEGF is overexpressed in human cancers, including highly vascularized and infiltrative brain tumors. In our previous study of seven glioma cell lines, VEGF expression levels correlated with blood vessel density and tumorigenicity, and U251 MG and NG-1 cells were recognized as low-tumorigenic glioma cell lines. We hypothesized that low-tumorigenic cells can become highly tumorigenic when high levels of VEGF are expressed. To test this hypothesis, we constructed VEGF expression vectors containing 564 bp or 696 bp of VEGF(121) or VEGF(165) cDNA, respectively, and transfected them into U251 MG and NG-1 cells. In comparison to parental cells, the 20 VEGF-expressing clones examined had on average 8-10-fold more VEGF mRNA and 12-88-fold more secreted VEGF proteins. Four VEGF-overexpressing clones (U251 MG/V121-C2, U251 MG/V165-C3, NG-1/V121-C6, and NG-1/V165-C3) were selected for additional study. As VEGF production increased with population growth, U251 MG/V121-C2 and U251 MG/V165-C3 cells accumulated 47.9 and 22.0 ng of VEGF during a 5-day culture of 10(4) cells, a 313- and 144-fold overexpression when compared with that in parental U251 MG cells. NG-1/V121-C6 and NG-1/V165-C3 cells secreted 30.4 and 9.4 ng of VEGF, respectively, or 138- and 43-fold more than did the parental NG-1 cells. Subcutaneous implantation of the VEGF-overexpressing U251 MG cells into nude mice caused huge, soft hemorrhagic tumors to form, whereas controls maintained very small tumors. Intracranial implantation of the VEGF-overexpressing cell lines significantly shortened survival of the mice when compared with controls, and it caused formation of solid brain tumors with variable sized hemorrhages, whereas the controls had no apparent brain tumors. Tumorigenicity of U251 MG cells was synergized by co-overexpression of VEGF(121) and VEGF(165). In addition, VEGF(165) seemed to be more potent to the brain endothelium than was VEGF(121). More interestingly, except when an admixture of cells was implanted s.c., VEGF overexpression in NG-1 cells did not promote hemorrhagic tumor formation. These data suggested that a switch from a phenotype of low tumorigenicity to one of high tumorigenicity is possible when VEGF overexpression occurs, although other factors may also be required. PMID- 11912166 TI - Noninvasive imaging of spontaneous retinoblastoma pathway-dependent tumors in mice. AB - Identification of the critical pathways involved in tumorigenesis should ultimately lead to the design of better anticancer agents that target specific components of the disrupted pathways. Murine models of spontaneous cancer in which tumor formation is dependent on defined genetic alterations provide a powerful test system for evaluating the therapeutic efficacy of pathway-specific antineoplastics. We have generated a conditional mouse model for retinoblastoma dependent sporadic cancer that permits noninvasive monitoring of pituitary tumor development in live animals via in vivo bioluminescence imaging of luciferase expression. We show that the high sensitivity of bioluminescence imaging can be used for noninvasive detection of luciferase expression in pituitary glands from tumor-free animals and for in vivo quantitation of tumor burden over a large dynamic range. This mouse model permits longitudinal monitoring of tumor onset, progression, and response to therapy and may be used effectively for testing cancer prevention and treatment strategies based on therapeutics that specifically target the retinoblastoma pathway. PMID- 11912167 TI - Identification of hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein I as a biomarker for pancreatic ductal adenocarcinoma by protein biochip technology. AB - New biomarkers of pancreatic adenocarcinoma are needed to improve the early detection of this deadly disease. We performed surface enhanced laser desorption ionization (SELDI) mass spectrometry using ProteinChip technology (Ciphergen Biosystems, Fremont, CA) to screen for differentially expressed proteins in pancreatic juice. Pancreatic juice samples obtained from patients undergoing pancreatectomy for pancreatic adenocarcinoma were compared with juice samples from patients with other pancreatic diseases. We identified a peak approximately 16,570 daltons present in the pancreatic juice from 10/15 (67%) of the patients with pancreatic adenocarcinoma and in the pancreatic juice from 1/7 (17%) of the patients with other pancreatic diseases. Using a ProteinChip immunoassay, we identified this differentially expressed protein as hepatocarcinoma-intestine pancreas/pancreatitis-associated-protein I (HIP/PAP-I), a protein released from pancreatic acini during acute pancreatitis and overexpressed in hepatocellular carcinoma. We then quantified by ELISA the pancreatic juice HIP/PAP-I levels in 43 patients (28 with pancreatic adenocarcinoma, 15 with other pancreatic diseases) and the serum HIP/PAP-I levels in 98 patients (53 with pancreatic adenocarcinoma, 45 with other pancreatic diseases or healthy individuals). HIP/PAP-I levels were significantly higher in both the pancreatic juice (P < 0.001) and in the serum (P < 0.001) of patients with pancreatic adenocarcinoma compared with the control group. HIP/PAP-I levels were approximately 1000-fold higher in pancreatic juice compared with serum and the magnitude of the difference between the pancreatic adenocarcinoma group and the control group was greater in the pancreatic juice samples (143.75 +/- 235.52 microg/ml versus 6.04 +/- 7.59 microg/ml) than in the serum samples (99.96 +/- 140.66 ng/ml versus 35.25 +/- 28.44 ng/ml). In our study, patients with pancreatic juice HIP/PAP-I levels > or= 20 microg/ml were 21.9 times (95% confidence interval, 3.5-136.5; P < 0.001) more likely to have pancreatic adenocarcinoma than patients with levels <20 microg/ml. Immunolabeling of tissue sections revealed that the HIP/PAP-I protein was strongly expressed in acini adjacent to the invasive adenocarcinoma, but it was only rarely (1/30; 3%) expressed in the neoplastic epithelium, which suggests that the main source of HIP/PAP-I release in the pancreatic juice is acini. This low level of HIP/PAP-I expression in pancreatic adenocarcinoma was confirmed by reverse transcription-PCR: only 1 (5%) of 19 pancreatic cancer cell lines expressed HIP/PAP-I transcripts. Taken together, these data suggest that pancreatic juice measurement of HIP/PAP-I may help to identify patients with pancreatic adenocarcinoma. PMID- 11912168 TI - DNA damage is able to induce senescence in tumor cells in vitro and in vivo. AB - Often the use of cytotoxic drugs in cancer therapy results in stable disease rather than regression of the tumor, and this is typically seen as a failure of treatment. We now show that DNA damage is able to induce senescence in tumor cells expressing wild-type p53. We also show that cytotoxics are capable of inducing senescence in tumor tissue in vivo. Our results suggest that p53 and p21 play a central role in the onset of senescence, whereas p16(INK4a) function may be involved in maintaining senescence. Thus, like apoptosis, senescence appears to be a p53-induced cellular response to DNA damage and an important factor in determining treatment outcome. PMID- 11912169 TI - Strategies for antigen loading of dendritic cells to enhance the antitumor immune response. AB - Dendritic cells (DCs) primed with tumor antigens can effectively mediate the regression of a variety of established solid malignancies in both murine and human models. Despite such clinical efficacy, the optimal means of DC priming is unknown. The goal of this study was to compare three methods of tumor preparation: irradiation, boiling, or freeze thaw lysis for DC priming. Mouse bone marrow-derived DCs were loaded with defined ratios of E.G7 tumor cells expressing a model tumor antigen, OVA. Sensitized DCs were used for stimulation of OVA-specific CTLs derived from OT-1 T-cell receptor transgenic mice. IFN-gamma release, determined by ELISA at 24 and 48 h, was used to assess the expression of antigens by DCs. DCs loaded with irradiated tumors were effective stimulators for OT-1 CTLs, whereas DCs stimulated with freeze-thawed or boiled tumors did not stimulate IFN-gamma production. Freeze-thaw lysis appeared to inhibit CTL activity in vitro and in two of three cases, this effect was not overcome by the addition of OVA. The ability to load irradiated tumor cells was reproduced in two analogous human melanoma models using melanoma cell lines expressing gp100 and CTL clones specific for a gp100 melanoma antigen. Consistent with the in vitro data, only DC/irradiated tumor vaccines were effective in preventing or delaying outgrowth of E.G7 and a poorly immunogenic murine squamous cell carcinoma (SCCVII), on local tumor challenge. These data demonstrate that the method of tumor cell preparation clearly influences the ability of DCs to present antigen to T cells. Correlation of in vitro data with the generation of protective immunity in vivo suggests the utility of irradiated tumor-primed DCs as a means to generate protective immunity in patients with solid malignancies. PMID- 11912170 TI - Melanoma tumors acquire a new phospholipid metabolism phenotype under cystemustine as revealed by high-resolution magic angle spinning proton nuclear magnetic resonance spectroscopy of intact tumor samples. AB - N'-[2-chloroethyl]-N[2-(methylsulfonyl) ethyl]-N'-nitrosourea (cystemustine),a chloroethylnitrosourea antineoplastic drug, provokes cellular proliferation inhibition and redifferentiation, but there was no cell death in B16 melanoma tumors. Because the phospholipid (Plp) metabolism is tightly involved in tumor growth regulation and tumor cell survival, we tested the hypothesis that melanoma tumors undergo adaptive Plp metabolism changes to survive treatment. Measurements of Plp derivatives were performed using a novel proton nuclear magnetic resonance Spectroscopy application using magic angle spinning on intact tumor tissue samples. Phosphatidylcholine levels were obtained from one-dimensional spectra, and relative levels of choline- and ethanolamine-containing compounds were derived from two-dimensional spectra (total correlation spectroscopy sequence). Two major findings emerged from this study: (a) during tumor growth inhibition, there was a transient accumulation of choline, glycerophosphocholine, and glycerophosphoethanolamine and a sustained increase in phosphocholine and phosphoethanolamine, whereas phosphatidylcholine levels remained unchanged; and (b) during tumor growth recovery, only phosphocholine and phosphoethanolamine remained elevated. Therefore, cystemustine-treated B16 melanoma tumors acquire a new Plp metabolism phenotype, a mechanism that could participate in tumor cell redifferentiation and/or survival. PMID- 11912171 TI - Fibroblast growth factor receptor 1 phosphotyrosine 766: molecular target for prevention of progression of prostate tumors to malignancy. AB - Dissection of processes that promote the slow progression to malignancy from those that drive the malignant phenotype, once acquired, is important for identification of molecular targets for rational design of dietary and pharmaceutical intervention to hold premalignant cancer in check. In adult parenchymal organs, fibroblast growth factor receptor (FGFR) kinase isotypes are partitioned between stroma and epithelium, respectively, and mediate communication between the two compartments to maintain organ homeostasis. The ectopic appearance of stromal FGFR1 is a hallmark of epithelial cells from model transplantable rat prostate tumors that progress to malignancy. Here we show that, despite the fact that it is transcriptionally active, the appearance of FGFR1 in nonmalignant prostate tumor epithelial cells at first does not drive cell proliferation or support a malignant phenotype. These properties develop over time with proliferative aging of the cell population coincident with FGFR1 dependent activation of the mitogen-activated protein kinase signaling pathway. Phospholipase Cgamma-interactive phosphotyrosine 766 of FGFR1 is required for the age-dependent acquisition of the proliferative response to FGFR1, although it appears not to be required for the mitogenic response. Although of little utility in late-stage therapy, this suggests that pathways linked to FGFR1 tyrosine 766 may be specific targets for prevention of progression of latent nonmalignant tumors to the life-threatening malignant state. PMID- 11912172 TI - Bag1 proteins regulate growth and survival of ZR-75-1 human breast cancer cells. AB - Bag1 proteins bind heat shock protein M(r) 70,000 (Hsp 70) family molecular chaperones and regulate diverse pathways involved in cell proliferation, apoptosis, and stress responses. Four isoforms of Bag1 can be produced from a single gene in humans, including a nuclear-targeted long version (Bag1L)and a shorter cytosolic isoform (Bag1). Because overexpression of Bag1and Bag1L has been reported in breast cancers, we explored the effects of Bag1 and Bag1L on the growth of ZR-75-1 human breast cancer cells cultured in vitro and in tumor xenograft models using immunocompromised mice. Cells stably transfected with expression plasmids encoding either Bag1 or Bag1L displayed comparable rates of growth in cultures containing 10% serum, compared with control-transfected ZR-75 1 cells. In contrast, ZR-75-1 cells stably expressing mutants of Bag1 or Bag1L, which lack the COOH-terminal domain (DeltaC) required for heat shock protein M(r) 70,000 binding, displayed retarded growth rates. When cultured without serum, the viability of control-transfected, as well as Bag1DeltaC- and Bag1LDeltaC expressing, cells declined with time, whereas Bag1- and Bag1L-overexpressing ZR 75-1 cells survived for over a week in culture. Caspase protease activation induced by serum deprivation was also prevented by stable expression of either Bag1 or Bag1L in ZR-75-1 cells. In addition, sensitivity to anchorage dependence was restored partially in ZR-75-1 cells expressing dominant-negative Bag1DeltaC and Bag1LDeltaC. In tumor xenograft studies involving injection of ZR-75-1 cells into mammary fat pads of female nu/nu mice, ZR-75-1 cells expressing Bag1 or Bag1L formed 1.4-1.6-fold larger tumors compared with control-transfected cells, whereas tumors formed by Bag1DeltaC- and Bag1LDeltaC-expressing cells grew very slowly and reached sizes < one-third of tumors generated by Neo-control ZR-75-1 cells. Altogether, these findings demonstrate that Bag1 and Bag1L provoke similar changes in breast cancer cell growth and survival and suggest that interference with Bag1 or Bag1L function might be a useful strategy for opposing breast cancer. PMID- 11912173 TI - Regulation of matrix metalloproteinase-9 (MMP-9) by translational efficiency in murine prostate carcinoma cells. AB - Expression of increased levels of matrix metalloproteinase-9 (MMP-9) has been implicated in tumor progression and angiogenesis. Much of our knowledge of the controls of MMP-9 levels has focused on transcription. Here we show that MMP-9 levels are also controlled by translational efficiency in murine prostate carcinoma cells. The murine prostate carcinoma cells 148-1,LMD and 148-1,PA were derived from a single mouse that had been implanted with urogenital sinus transformed by ras and myc. 148-1,PA secretes little MMP-9 yet has equivalent amounts of MMP-9 mRNA as the cell line 148-1,LMD that secretes substantially more. Infection with a retroviral vector for murine MMP-9 led to more expression of MMP-9 in both cases, but the differential remained. Human MMP-9 is equally expressed in both cells after infection with a vector for human MMP-9 indicating that the effect is species-specific. Pulse chase analysis revealed that MMP-9 was synthesized more rapidly in the 148-1,LMD cells than in the 148-1,PA cells. Markedly more MMP-9 mRNA was associated with polysomes in the cell line synthesizing more MMP-9. These results indicate regulation of MMP-9 synthesis at the level of translational efficiency. PMID- 11912174 TI - Prevention of irradiation-induced glioma cell invasion by temozolomide involves caspase 3 activity and cleavage of focal adhesion kinase. AB - Sublethal doses of irradiation enhance the invasiveness of human malignantglioma cells. This can be inhibited by subtoxic concentrations of temozolomide (TMZ) but not by lomustine. Antagonism of irradiation-induced motility by TMZ is associated with the prevention of irradiation-induced alpha(v)beta(3)-integrin, matrix metalloproteinase-2 and MT1-matrix metalloproteinase-expression. Irradiation induces focal adhesion kinase (FAK) activation by phosphorylation, whereas TMZ promotes FAK cleavage. Inhibition of caspases prevents TMZ-induced FAK processing and restores the promigratory effect of irradiation, suggesting that the resistance of glioma cells to irradiation-induced caspase processing may determine the invasive responses of glioma cells to irradiation. In contrast, DAOY medulloblastoma cells, which respond with caspase activation to irradiation alone, do not show enhanced invasiveness when irradiated. PMID- 11912175 TI - Chemotherapy induces lytic EBV replication and confers ganciclovir susceptibility to EBV-positive epithelial cell tumors. AB - EBV is an oncogenic herpesvirus associated with a number of human malignancies. The consistent presence of the EBV genome in certain tumors offers the potential for novel EBV-targeted therapies. EBV can infect cells in either a latent or lytic form. Here we demonstrate that a variety of chemotherapeutic agents, including cis-platinum, 5-fluorouracil (5-FU), and taxol, induce the switch from the latent to lytic form of EBV infection in tumor cells. This effect requires the protein kinase C delta, phosphatidylinositol 3'-kinase, and p38 stress mitogen-activated protein kinase signaling pathways but not caspase 3 activation. Because the lytic but not latent form of EBV infection converts the cytotoxic prodrug, ganciclovir (GCV), into its active form, we examined whether the combination of GCV and chemotherapy is more effective than chemotherapy alone for killing EBV-positive tumor cells. GCV significantly enhanced the ability of 5-FU and cis-platinum to kill EBV-positive, but not EBV-negative, gastric carcinoma cells in vitro. Most importantly, the combination of GCV and 5-FU (or GCV and cis platinum) was much more effective in the treatment of EBV-positive nasopharyngeal carcinomas passaged in nude mice than either agent alone. These data suggest that GCV enhances the efficacy of conventional chemotherapy for the treatment of EBV positive epithelial cell tumors. PMID- 11912176 TI - STAT6 as an asthma candidate gene: polymorphism-screening, association and haplotype analysis in a Caucasian sib-pair study. AB - The human signal transducer and activator of transcription 6 (STAT6) gene represents one of the most promising candidate genes for asthma and other inflammatory diseases on the chromosomal region 12q13-q24. Therefore we screened all 23 exons, including parts of the neighbouring introns, as well as the promoter region for common polymorphisms and tested them for linkage/association with asthma and related traits (total serum IgE level, eosinophil cell count and SLOPE of the dose-response curve after bronchial challenge) in a Caucasian sib pair study (108 families with at least two affected children). We could identify 13 single nucleotide polymorphisms (SNPs), which are all non-coding. A recently described dinucleotide (GT) repeat in exon 1 was also examined. Besides the confirmation of the four alleles described elsewhere we could identify a new one, named allele A5. Neither the SNPs nor the GT repeat showed linkage/association to asthma. Two intronic SNPs and one SNP in the 3'untranslated region of the gene showed weak association to total IgE levels (P = 0.0200, 0.0260 and 0.0280, respectively), whereas a significant association was found between a SNP in intron 18 and an increase in total IgE levels (P = 0.0070). However, the most promising effect was seen between allele A4 of the GT repeat polymorphism and an increase in eosinophil cell count (P = 0.0010). From these findings we conclude that the human STAT6 gene is rather involved in the development of eosinophilia and changes in total IgE levels than contributing to the pathogenesis of asthma. PMID- 11912177 TI - Genome-wide linkage analysis of severe, early-onset chronic obstructive pulmonary disease: airflow obstruction and chronic bronchitis phenotypes. AB - Familial aggregation of chronic obstructive pulmonary disease (COPD) has been demonstrated, but linkage analysis of COPD-related phenotypes has not been reported previously. An autosomal 10 cM genome-wide scan of short tandem repeat (STR) polymorphic markers was analyzed for linkage to COPD-related phenotypes in 585 members of 72 pedigrees ascertained through severe, early-onset COPD probands without severe alpha1-antitrypsin deficiency. Multipoint non-parametric linkage analysis (using the ALLEGRO program) was performed for qualitative phenotypes including moderate airflow obstruction [forced expiratory volume at one second (FEV(1)) < 60% predicted, FEV(1)/FVC < 90% predicted], mild airflow obstruction (FEV(1) < 80% predicted, FEV(1)/FVC < 90% predicted) and chronic bronchitis. The strongest evidence for linkage in all subjects was observed at chromosomes 12 (LOD = 1.70) and 19 (LOD = 1.54) for moderate airflow obstruction, chromosomes 8 (LOD = 1.36) and 19 (LOD = 1.09) for mild airflow obstruction and chromosomes 19 (LOD = 1.21) and 22 (LOD = 1.37) for chronic bronchitis. Restricting analysis to cigarette smokers only provided increased evidence for linkage of mild airflow obstruction and chronic bronchitis to several genomic regions; for mild airflow obstruction in smokers only, the maximum LOD was 1.64 at chromosome 19, whereas for chronic bronchitis in smokers only, the maximum LOD was 2.08 at chromosome 22. On chromosome 12p, 12 additional STR markers were genotyped, which provided additional support for an airflow obstruction locus in that region with a non parametric multipoint approach for moderate airflow obstruction (LOD = 2.13) and mild airflow obstruction (LOD = 1.43). Using a dominant model with the STR markers on 12p, two point parametric linkage analysis of all subjects demonstrated a maximum LOD score of 2.09 for moderate airflow obstruction and 2.61 for mild airflow obstruction. In smokers only, the maximum two point LOD score for mild airflow obstruction was 3.14. These observations provide suggestive evidence that there is a locus on chromosome 12p which contributes to susceptibility to early-onset COPD. PMID- 11912179 TI - Disruption of a novel MFS transporter gene, DIRC2, by a familial renal cell carcinoma-associated t(2;3)(q35;q21). AB - Previously, we described a family with a significantly increased predisposition for renal cell cancer co-segregating with a t(2;3)(q35;q21) chromosomal translocation. Several primary tumors of the clear cell type from different family members were analyzed at a molecular level. Loss of the derivative chromosome 3 was consistently found. In addition, different somatic Von Hippel Lindau (VHL) gene mutations were observed in most of the tumors analyzed, even within the same patient. Based on these results a multistep tumorigenesis model was proposed in which (non-disjunctional) loss of the derivative chromosome 3 represents an early event and somatic mutation of the VHL gene represents a late event related to tumor progression. More recently, however, we noted that these two anomalies were absent in at least one early-stage tumor sample that we tested. Similar results were obtained in another family with renal cell cancer and t(3;6)(q12;q15), thus suggesting that another genetic event may precede these two oncogenetic steps. We speculate that deregulation of a gene(s) located at or near the translocation breakpoint may act as such. In order to identify such genes, a detailed physical map encompassing the 3q21 breakpoint region was constructed. Through a subsequent positional cloning effort we found that this breakpoint targets a hitherto unidentified gene, designated DIRC2 (disrupted in renal cancer 2). Computer predictions of the putative DIRC2 protein showed significant homology to different members of the major facilitator superfamily (MFS) of transporters. Based on additional DIRC2 expression and mutation analyses, we propose that the observed gene disruption may result in haplo insufficiency and, through this mechanism, in the onset of tumor growth. PMID- 11912178 TI - Early phenotypes that presage late-onset neurodegenerative disease allow testing of modifiers in Hdh CAG knock-in mice. AB - In Huntington's disease (HD), CAG repeats extend a glutamine tract in huntingtin to initiate the dominant loss of striatal neurons and chorea. Neuropathological changes include the formation of insoluble mutant N-terminal fragment, as nuclear/neuropil inclusions and filter-trap amyloid, which may either participate in the disease process or be a degradative by-product. In young Hdh knock-in mice, CAGs that expand the glutamine tract in mouse huntingtin to childhood-onset HD lengths lead to nuclear accumulation of full-length mutant huntingtin and later accumulation of insoluble fragment. Here we report late-onset neurodegeneration and gait deficits in older Hdh(Q111) knock-in mice, demonstrating that the nuclear phenotypes comprise early stages in a disease process that conforms to genetic and pathologic criteria determined in HD patients. Furthermore, using the early nuclear-accumulation phenotypes as surrogate markers, we show in genetic experiments that the disease process, initiated by full-length mutant protein, is hastened by co-expression of mutant fragment; therefore, accrual of insoluble-product in already compromised neurons may exacerbate pathogenesis. In contrast, timing of early disease events was not altered by normal huntingtin or by mutant caspase-1, two proteins shown to reduce inclusions and glutamine toxicity in other HD models. Thus, potential HD therapies in man might be directed at different levels: preventing the disease initiating mechanism or slowing the subsequent progression of pathogenesis. PMID- 11912180 TI - WT1 is a key regulator of podocyte function: reduced expression levels cause crescentic glomerulonephritis and mesangial sclerosis. AB - Glomerular disease is one of the most common causes of end-stage renal failure. Increasing evidence suggests that these glomerulopathies are frequently caused by primary lesions in the renal podocytes. One of the major consequences of podocyte lesions is the accumulation of mesangial matrix in the glomerular basement membrane, a process called glomerulosclerosis. Mesangial sclerosis is one of the most consistent findings in Denys-Drash patients and can be caused by dominant mutations in the Wilms' tumor 1 gene (WT1). The underlying mechanism, however, is poorly understood. WT1 is expressed in the podocytes throughout life, but its function in this cell type is unknown. Combining Wt1-knockout and inducible yeast artificial chromosome transgenic mouse models, we demonstrate that reduced expression levels of WT1 result in either crescentic glomerulonephritis or mesangial sclerosis depending on the gene dosage. Strikingly, the two podocyte specific genes nphs1 and podocalyxin are dramatically downregulated in mice with decreased levels of Wt1, suggesting that these two genes act downstream of Wt1. Taken together, our data provide genetic evidence that reduced levels of Wt1 are responsible for the pathogenesis of two distinct renal diseases and offer a molecular explanation for the increased occurrence of glomerulosclerosis in patients with WAGR syndrome. PMID- 11912181 TI - Mapping of an autoimmunity susceptibility locus (AIS1) to chromosome 1p31.3 p32.2. AB - Generalized vitiligo is a common autoimmune disorder in which patchy loss of skin and hair pigmentation results from loss of pigment-forming melanocytes from the involved regions. Vitiligo occurs with a frequency of about 1% in most populations, and is highly associated with other autoimmune disorders, particularly Hashimoto thyroiditis. Most cases of vitiligo are sporadic, although some cases cluster in families, and the disorder is thought to be oligogenic in origin. We have studied a large family cluster in which vitiligo and Hashimoto thyroiditis occur in numerous individuals. A whole-genome scan of 24 family members, including 14 affected with autoimmune disease, showed significant linkage of an oligogenic autoimmune susceptibility locus, termed AIS1, to a 14.4 cM interval in 1p31.3-p32.2. A two-locus analysis of Hashimoto thyroiditis in family members segregating an AIS1 susceptibility allele showed suggestive linkage to markers in chromosome 6p22.3-q14.1, in a region spanning both the major histocompatibility complex and AITD1, a susceptibility locus for autoimmune thyroid disease. Our results indicate that the 1p AIS1 locus is associated with susceptibility to autoimmunity, particularly vitiligo, in this family, and that a chromosome 6 locus, most likely AITD1, may mediate the occurrence of Hashimoto's thyroiditis in AIS1-susceptible family members. PMID- 11912182 TI - Classification of common conserved sequences in mammalian intergenic regions. AB - Comparisons between orthologous intergenic regions of related genomes reveal numerous hits, i.e. pairs of relatively short highly similar sequences that evolved slowly, perhaps due to selective constraint. We analyzed and classified 2638 hits found within 100 pairs of complete, orthologous intergenic regions of human and murine genomes. We identified all common fragments of hits that align well with many other hits and constructed their classification. Our analysis revealed 20 abundant classes each containing 10 or more fragments. Fragments of the same class may perform the same function, e.g. bind a particular protein. Ten of the abundant classes apparently correspond to known functional consensuses, whereas others may represent novel conserved sites. Thus, large-scale comparative analysis of slowly evolving intergenic sequences can provide valuable insights into their function. PMID- 11912183 TI - The expression of a new variant of the pro-apoptotic molecule Bax, Baxpsi, is correlated with an increased survival of glioblastoma multiforme patients. AB - Pro- and anti-apoptotic members of the BCL-2 family play a central role in the implementation of apoptosis. Bax, a pro-apoptotic member of this family, has as such been considered as a potential tumor suppressor. Here, we have examined the expression of Bax in 55 patients with glioblastoma multiforme (GBM), the most common and aggressive form of brain tumors. We report on the existence of a new form of Bax, present in 24% of the patients, which we called Baxpsi. Baxpsi is a N-terminal truncated form of Bax which results from a partial deletion of the exon 1 of Bax gene. Baxpsi and the wild-type form, Baxalpha, are encoded by distinct mRNAs, both of which are present in normal tissues. Glial tumors express either Baxalpha or Baxpsi proteins, an apparent consequence of an exclusive transcription of the corresponding mRNAs. The latter feature could be partially linked to distinct methylation profiles of Bax gene in these tumors. The Baxpsi protein is preferentially localized to mitochondria and is a more powerful inducer of apoptosis than Baxalpha. Baxpsi tumors exhibit a slow proliferation in Swiss nude mice and this feature can be circumvented by the co-expression of the Bcl-2 transgene, the functional antagonist of Bax. More importantly, the expression of Baxpsi correlates with a longer survival in patients (18 months versus 10 months for Baxalpha patients). Thus, our results provide the first indication of a beneficial involvement of a variant of the pro-apoptotic protein Bax in tumor progression. PMID- 11912184 TI - Linkage analysis of anorexia nervosa incorporating behavioral covariates. AB - Eating disorders, such as anorexia nervosa (AN) and bulimia nervosa (BN), have genetic and environmental underpinnings. To explore genetic contributions to AN, we measured psychiatric, personality and temperament phenotypes of individuals diagnosed with eating disorders from 196 multiplex families, all accessed through an AN proband, as well as genotyping a battery of 387 short tandem repeat (STR) markers distributed across the genome. On these data we performed a multipoint affected sibling pair (ASP) linkage analysis using a novel method that incorporates covariates. By exploring seven attributes thought to typify individuals with eating disorders, we identified two variables, drive-for thinness and obsessionality, which delimit populations among the ASPs. For both of these traits, or covariates, there were a cluster of ASPs who have high and concordant values for these traits, in keeping with our expectations for individuals with AN, and other clusters of ASPs who did not meet those expectations. When we incorporated these covariates into the ASP linkage analysis, both jointly and separately, we found several regions of suggestive linkage: one close to genome-wide significance on chromosome 1 (at 210 cM, D1S1660; LOD = 3.46, P = 0.00003), another on chromosome 2 (at 114 cM, D2S1790; LOD = 2.22, P = 0.00070) and a third region on chromosome 13 (at 26 cM, D13S894; LOD = 2.50, P = 0.00035). By comparing our results to those implemented using more standard linkage methods, we find the covariates convey substantial information for the linkage analysis. PMID- 11912186 TI - Sequence dependent instability of mononucleotide microsatellites in cultured mismatch repair proficient and deficient mammalian cells. AB - We have measured the mutation rates of G(17) and A(17) repeat sequences in cultured mammalian cells with and without mismatch repair and have compared these rates to those of a (CA)(17) repeat sequence. Plasmids containing microsatellites that disrupt the reading frame of a downstream neomycin-resistance gene were introduced into the cells by transfection and revertants were selected using the neomycin analog G418. Comparison of mutation rates within cell lines showed that the mutation rates of A(17) and (CA)(17) sequences were similar in the mismatch repair proficient cells, but the mutation rate of G(17) was significantly higher than that of either A(17) or (CA)(17). In the mismatch repair deficient cells, the G(17) and (CA)(17) mutation rates were similar and were significantly higher than the A(17) rate. PCR analysis of the mutants showed that 1 bp insertions predominated in both mononucleotide repeats in the mismatch repair proficient cells; in mismatch repair deficient cells, 2 bp deletions were the most common mutation in the A(17) sequence, but 1 bp insertions and 2 bp deletions were equally represented in the G(17) sequence. These results indicate that a G(17) repeat is less stable than an A(17) repeat in both mismatch repair proficient and mismatch repair deficient mammalian cells. This observation implies that the replication fidelity is lower in G(17) repeats. PMID- 11912185 TI - The gene for the muted (mu) mouse, a model for Hermansky-Pudlak syndrome, defines a novel protein which regulates vesicle trafficking. AB - The muted (mu) mouse is a model for Hermansky-Pudlak Syndrome (HPS), an inherited disorder of humans causing hypopigmentation, hemorrhaging and early death due to lung abnormalities. The mu gene regulates the synthesis of specialized mammalian organelles such as melanosomes, platelet dense granules and lysosomes. Further, balance defects indicate that it controls the synthesis of otoliths of the inner ear. The mu gene has been identified by a positional/candidate approach involving large mouse interspecific backcrosses. It encodes a novel ubiquitously expressed transcript, specifying a predicted 185 amino acid protein, whose expression is abrogated in the mu allele which contains an insertion of an early transposon (ETn) retrotransposon. Expression is likewise expected to be lost in the mu( J) allele which contains a deletion of a single base pair within the coding region. The presence of structurally aberrant melanosomes within the eyes of mutant mice together with localization of the muted protein within vesicles in both the cell body and dendrites of transfected melan-a melanocytes emphasizes the role of the mu gene in vesicle trafficking. The mu gene is present only in mice and humans among analyzed genomes. As is true for several other recently identified mouse HPS genes, the mu gene is absent in lower eukaryotes. Therefore, the mu gene is a member of the novel gene set that has evolved in higher eukaryotes to regulate the synthesis/function of highly specialized subcellular organelles such as melanosomes and platelet dense granules. PMID- 11912187 TI - Loss of function of axonemal dynein Mdnah5 causes primary ciliary dyskinesia and hydrocephalus. AB - Primary ciliary dyskinesia (PCD), also known as Kartagener's syndrome, is a human syndrome that results from ciliary dysfunction. This syndrome is characterized by recurrent respiratory infections, situs inversus and infertility. In some cases, hydrocephalus is also observed. We have characterized an insertional mutation in a mouse axonemal dynein heavy chain gene (Mdnah5) that reproduces most of the classical features of PCD, including recurrent respiratory infections, situs inversus and ciliary immotility. These mice also suffer from hydrocephalus and die perinatally. Electron microscopic studies demonstrate the loss of axonemal outer arms. These results show that mutations in Mdnah5 are a primary cause of PCD and provide direct evidence that mutations in an axonemal dynein can cause hydrocephalus. Mutations in the human DNAH5 have recently been identified in PCD patients. Comparison of the mouse model and the human data suggests that the degree of ciliary dysfunction is causally related to the severity of human PCD, particularly the presence of hydrocephalus. PMID- 11912188 TI - Phosphorylation-dependent cellular localization and thermoprotective role of heat shock protein 25 in hippocampal progenitor cells. AB - The present study examined phosphorylation-dependent cellular localization and the thermoprotective role of heat shock protein (HSP) 25 in hippocampal HiB5 cells. HSP25 was induced and phosphorylated by heat shock (at 43 degrees C for 3 h). HSP25, which was located in the cytoplasm in the normal condition, translocated into the nucleus after the heat shock. Transfection experiments with hsp27 mutants in which specific serine phosphorylation residues (Ser(78) and Ser(82)) were substituted with alanines or aspartic acids showed that phosphorylation of HSP27 is accompanied by its nuclear translocation. Phosphorylation of mitogen-activated protein kinases (MAPKs) such as p38 MAPK and ERK was markedly increased by the heat shock, and SB203580 (a p38 MAPK kinase inhibitor) and/or PD098059 (a MEK inhibitor) inhibited the phosphorylation of HSP25, indicating that p38 MAPK and ERK are upstream regulators of HSP25 phosphorylation in the heat shock condition. In the absence of heat shock, actin filament stability was not affected by SB203580 and/or PD098059. Heat shock caused disruption of the actin filament and cell death when phosphorylation of HSP25 was inhibited by SB203580 and/or PD098059. In addition, actin filament was more stable in Asp(78,82)-hsp27 (mimics the phosphorylated form) transfected HiB5 cells than in the normal and Ala(78,82)-hsp27 (nonphosphorylative form) transfected cells. In accordance with actin filament stability, the survival rate against the heat shock increased markedly in Asp(15,78,82)-hsp27 expressing HiB5 cells but decreased in Ala(15,78,82)-hsp27 expressing cells. These results support the idea that phosphorylation of HSP25 is critical for the maintenance of actin filament and enhancement of thermoresistance. Interestingly, HSP25 was dephosphorylated and returned to cytoplasm in a recovery time-dependent manner. This phenomenon was accompanied by an increment of apoptotic cell death as determined by nuclear and DNA fragmentation and fluorescence-activated cell sorter analysis. These results suggest that nuclear-translocated HSP25 might function to protect nuclear structure, thereby preventing apoptotic cell death. PMID- 11912190 TI - Attenuation of protein kinase C and cAMP-dependent protein kinase signal transduction in the neurogranin knockout mouse. AB - Neurogranin (Ng) is a brain-specific, postsynaptically located protein kinase C (PKC) substrate, highly expressed in the cortex, hippocampus, striatum, and amygdala. This protein is a Ca(2+)-sensitive calmodulin (CaM)-binding protein whose CaM-binding affinity is modulated by phosphorylation and oxidation. To investigate the role of Ng in neural function, a strain of Ng knockout mouse (KO) was generated. Previously we reported (Pak, J. H., Huang, F. L., Li, J., Balschun, D., Reymann, K. G., Chiang, C., Westphal, H., and Huang, K.-P. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 11232-11237) that these KO mice displayed no obvious neuroanatomical abnormality, but exhibited deficits in learning and memory and activation of Ca(2+)/CaM-dependent protein kinase II. In this report, we analyzed several downstream phosphorylation targets in phorbol 12-myristate 13 acetate- and forskolin-treated hippocampal slices from wild type (WT) and KO mice. Phorbol 12-myristate 13-acetate caused phosphorylation of Ng in WT mice and promoted the translocation of PKC from the cytosolic to the particulate fractions of both the WT and KO mice, albeit to a lesser extent in the latter. Phosphorylation of downstream targets, including mitogen-activated protein kinases, 90-kDa ribosomal S6 kinase, and the cAMP response element binding protein (CREB) was significantly attenuated in KO mice. Stimulation of hippocampal slices with forskolin also caused greater stimulation of protein kinase A (PKA) in the WT as compared with those of the KO mice. Again, phosphorylation of the downstream targets of PKA was attenuated in the KO mice. These results suggest that Ng plays a pivotal role in regulating both PKC- and PKA-mediated signaling pathways, and that the deficits in learning and memory of spatial tasks detected in the KO mice may be the result of defects in the signaling pathways leading to the phosphorylation of CREB. PMID- 11912189 TI - Interaction of Alzheimer's beta -amyloid precursor family proteins with scaffold proteins of the JNK signaling cascade. AB - We have isolated a novel protein based on its association with Drosophila APP like protein (APPL), a homolog of the beta-amyloid precursor protein (APP) that is implicated in Alzheimer's disease. This novel APPL-interacting protein 1 (APLIP1) contains a Src homology 3 domain and a phosphotyrosine interaction domain and is expressed abundantly in neural tissues. The phosphotyrosine interaction domain of APLIP1 interacts with a sequence containing GYENPTY in the cytoplasmic domain of APPL. APLIP1 is highly homologous to the carboxyl-terminal halves of mammalian c-Jun NH(2)-terminal kinase (JNK)-interacting protein 1b (JIP1b) and 2 (JIP2), which also contain Src homology 3 and phosphotyrosine interaction domains. The similarity of APLIP1 to JIP1b and JIP2 includes interaction with component(s) of the JNK signaling pathway and with the motor protein kinesin and the formation of homo-oligomers. JIP1b interacts strongly with the cytoplasmic domain of APP (APPcyt), as APLIP1 does with APPL, but the interaction of JIP2 with APPcyt is weak. Overexpression of JIP1b slightly enhances the JNK-dependent threonine phosphorylation of APP in cultured cells, but that of JIP2 suppresses it. These observations suggest that the interactions of APP family proteins with APLIP1, JIP1b, and JIP2 are conserved and play important roles in the metabolism and/or the function of APPs including the regulation of APP phosphorylation by JNK. Analysis of APP family proteins and their associated proteins is expected to contribute to understanding the molecular process of neural degeneration in Alzheimer's disease. PMID- 11912191 TI - The molecular basis of Src kinase specificity during vertebrate mesoderm formation. AB - Members of the Src family of non-receptor tyrosine kinases play a critical role in mesoderm formation in the frog, Xenopus laevis, acting as required mediators downstream of the fibroblast growth factor receptor. At least four members of this gene family, Src, Fyn, Yes, and Laloo, are expressed during early embryonic development. Ectopic expression of Laloo and Fyn, but not Src, induce mesoderm in ectodermal explants, indicating that these factors are non-redundant during early vertebrate development. Here we investigate the basis for the differential activity of the Src and Laloo kinases during mesoderm formation. We demonstrate that although both Src and Laloo physically interact with the substrate protein SNT-1/FRS2alpha only Laloo phosphorylates SNT-1, an event previously shown to be required for the activity of the latter and for mesoderm induction in vivo. We show that Src is enzymatically capable of stimulating mesoderm formation, as an activated Src construct both phosphorylates SNT-1 and induces mesoderm in explant cultures. However, a chimeric Laloo construct containing a Src C-terminal tail is inactive, suggesting that the early embryo contains a specific Laloo-activating, or Src-inactivating, factor. Finally, through further chimeric analysis, we provide evidence to suggest that differences in Laloo and Src activity are also mediated by the SH2, SH3, and kinase domains of these molecules. PMID- 11912192 TI - Pathway complexity of prion protein assembly into amyloid. AB - In vivo under pathological conditions, the normal cellular form of the prion protein, PrP(C) (residues 23-231), misfolds to the pathogenic isoform PrP(Sc), a beta-rich aggregated pathogenic multimer. Proteinase K digestion of PrP(Sc) leads to a proteolytically resistant core, PrP 27-30 (residues 90-231), that can form amyloid fibrils. To study the kinetic pathways of amyloid formation in vitro, we used unglycosylated recombinant PrP corresponding to the proteinase K-resistant core of PrP(Sc) and found that it can adopt two non-native abnormal isoforms, a beta-oligomer and an amyloid fibril. Several lines of kinetic data suggest that the beta-oligomer is not on the pathway to amyloid formation. The preferences for forming either a beta-oligomer or amyloid can be dictated by experimental conditions, with acidic pH similar to that seen in endocytic vesicles favoring the beta-oligomer and neutral pH favoring amyloid. Although both abnormal isoforms have high beta-sheet content and bind 1-anilinonaphthalene-8-sulfonate, they are dissimilar structurally. Multiple pathways of misfolding and the formation of distinct beta-sheet-rich abnormal isoforms may explain the difficulties in refolding PrP(Sc) in vitro, the need for a PrP(Sc) template, and the significant variation in disease presentation and neuropathology. PMID- 11912193 TI - Heparin amplifies platelet-derived growth factor (PDGF)- BB-induced PDGF alpha receptor but not PDGF beta -receptor tyrosine phosphorylation in heparan sulfate deficient cells. Effects on signal transduction and biological responses. AB - Platelet-derived growth factor (PDGF) induces mitogenic and migratory responses in a wide variety of cells, by activating specific receptor tyrosine kinases denoted the PDGF alpha- and beta-receptors. Different PDGF isoforms bind in a distinct manner to glycosaminoglycans, particularly heparan sulfate. In the present study, we show potentiation by exogenous heparin of PDGF-BB-induced PDGF alpha-receptor tyrosine phosphorylation in heparan sulfate-deficient Chinese hamster ovary (CHO) 677 cells. This effect was not seen for PDGF-AA treatment, and heparin lacked a potentiating effect on PDGF-BB stimulation of the PDGF beta receptor. Heparin did not affect the affinity of PDGF-BB binding for the PDGF receptors on CHO 677 cells. The PDGF-BB-stimulated PDGF alpha-receptor phosphorylation was enhanced in a dose-dependent fashion by heparin at low concentration. The effect was modulated by 2-O- and 6-O-desulfation of the polysaccharide. Maximal induction of PDGF alpha-receptor tyrosine phosphorylation (6-fold) in CHO 677 cells was achieved by treatment with a heparin decasaccharide, but shorter oligosaccharides consisting of four or more monosaccharide units were also able to augment PDGF alpha-receptor phosphorylation, albeit at higher concentrations. Heparin potentiated PDGF-BB induced activation of mitogen-activated protein kinase and protein kinase B (Akt) and allowed increased chemotaxis of the CHO 677 cells toward PDGF-BB. In conclusion, heparin modulates PDGF-BB-induced PDGF alpha-receptor phosphorylation and downstream signaling, with consequences for cellular responsiveness to the growth factor. PMID- 11912194 TI - Insulin receptor substrate 4 associates with the protein IRAS. AB - The insulin receptor substrates (IRSs) are key components in signaling from the insulin receptor, and consequently any proteins that interact with them are expected to participate in insulin signaling. In this study we have searched for proteins that interact with IRS-4 by identifying the proteins that coimmunoprecipitated with IRS-4 from human embryonic kidney 293 cells by microsequencing through mass spectrometry. A group of proteins was found. These included phosphatidylinositol 3-kinase, a protein previously identified as an IRS 4 interactor, and several proteins for which there was no previous evidence of IRS-4 association. One of these proteins, named IRAS, that had been found earlier in another context was examined in detail. The results from the overexpression of IRAS, where its amount was about the same as that of IRS-4, indicated that IRAS associated directly with IRS-4 and showed that the increased complexation of IRS 4 with IRAS did not alter the insulin-stimulated tyrosine phosphorylation of IRS 4 or the association of IRS-4 with phosphatidylinositol 3-kinase or Grb2. On the other hand, overexpression of IRAS enhanced IRS-4-dependent insulin stimulation of the extracellularly regulated kinase. The domains of IRAS and IRS-4 responsible for the association of these two proteins were identified, and it was shown that IRAS also associates with IRS-1, IRS-2, and IRS-3. PMID- 11912195 TI - Phosphorylation of varicella-zoster virus IE63 protein by casein kinases influences its cellular localization and gene regulation activity. AB - During the early phase of varicella-zoster virus (VZV) infection, Immediate Early protein 63 (IE63) is expressed rapidly and abundantly in the nucleus, while during latency, this protein is confined mostly to the cytoplasm. Because phosphorylation is known to regulate many cellular events, we investigated the importance of this modification on the cellular localization of IE63 and on its regulatory properties. We demonstrate here that cellular casein kinases I and II are implicated in the in vitro and in vivo phosphorylation of IE63. A mutational approach also indicated that phosphorylation of the protein is important for its correct cellular localization in a cell type-dependent fashion. Using an activity test, we demonstrated that IE63 was able to repress the gene expression driven by two VZV promoters and that phosphorylation of the protein was required for its full repressive properties. Finally, we showed that IE63 was capable of exerting its repressive activity in the cytoplasm, as well as in the nucleus, suggesting a regulation at the transcriptional and/or post-transcriptional level. PMID- 11912196 TI - Apolipoprotein E4 potentiates amyloid beta peptide-induced lysosomal leakage and apoptosis in neuronal cells. AB - We assessed the isoform-specific effects of apolipoprotein (apo) E on the response of Neuro-2a cells to the amyloid beta peptide (Abeta1-42). As determined by the intracellular staining pattern and the release of beta-hexosaminidase into the cytosol, apoE4-transfected cells treated with aggregated Abeta1-42 showed a greater tendency toward lysosomal leakage than neo- or apoE3-transfected cells. Abeta1-42 caused significantly greater cell death and more than 2-fold greater DNA fragmentation in apoE4-secreting than in apoE3-secreting or control cells. H2O2 or staurosporine enhanced cell death and apoptosis in apoE4-transfected cells but not in apoE3-transfected cells. A caspase-9 inhibitor abolished the potentiation of Abeta1-42-induced apoptosis by apoE4. Similar results were obtained with conditioned medium from cells secreting apoE3 or apoE4. Cells preincubated for 4 h with a source of apoE3 or apoE4, followed by removal of apoE from the medium and from the cell surface, still exhibited the isoform-specific response to Abeta1-42, indicating that the potentiation of apoptosis required intracellular apoE, presumably in the endosomes or lysosomes. Studies of phospholipid (dimyristoylphosphatidylcholine) bilayer vesicles encapsulating 5 (and-6)-carboxyfluorescein dye showed that apoE4 remodeled and disrupted the phospholipid vesicles to a greater extent than apoE3 or apoE2. In response to Abeta1-42, vesicles containing apoE4 were disrupted to a greater extent than those containing apoE3. These findings are consistent with apoE4 forming a reactive molecular intermediate that avidly binds phospholipid and may insert into the lysosomal membrane, destabilizing it and causing lysosomal leakage and apoptosis in response to Abeta1-42. PMID- 11912197 TI - Molecular mechanism of transforming growth factor (TGF)-beta1-induced glutathione depletion in alveolar epithelial cells. Involvement of AP-1/ARE and Fra-1. AB - Glutathione (GSH) is a ubiquitous antioxidant in lung epithelial cells and lung lining fluid. Transforming growth factor beta1 (TGF-beta1) is a pleiotropic cytokine involved in cellular proliferation and differentiation. The level of TGF beta1 is elevated in many chronic inflammatory lung disorders associated with oxidant/antioxidant imbalance. In this study, we show that TGF-beta1 depletes GSH by down-regulating expression of the enzyme responsible for its formation, gamma glutamylcysteine synthetase (gamma-GCS) and induces reactive oxygen species production in type II alveolar epithelial cells (A549). To investigate the molecular mechanisms of inhibition of glutathione synthesis, we employed reporters containing fragments from the promoter region of the gamma-GCS heavy subunit (h), the gene that encodes the catalytic subunit of gamma-GCS. We found that TGF-beta1 reduced the expression of the long gamma-GCSh construct ( 3802/GCSh-5'-Luc), suggesting that an antioxidant response element (ARE) may be responsible for mediating the TGF-beta1 effect. Interestingly, the electrophoretic mobility shift assay revealed that the DNA binding activity of both activator protein-1 (AP-1) and ARE was increased in TGF-beta1-treated epithelial cells. The gamma-GCSh ARE contains a perfect AP-1 site embedded within it, and mutation of this internal AP-1 sequence, but not the surrounding ARE, prevented DNA binding. Further studies revealed that c-Jun and Fra-1 dimers, members of the AP-1 family previously shown to exert a negative effect on phase II gene expression, bound to the ARE sequence. We propose a novel mechanism of gamma-GCSh down-regulation by TGF-beta1 that involves the binding of c-Jun and Fra-1 dimers to the distal promoter. The findings of this study provide important information, which may be used for the modulation of glutathione biosynthesis in inflammation. PMID- 11912198 TI - Oxidation of methionine 35 attenuates formation of amyloid beta -peptide 1-40 oligomers. AB - Amyloid plaques formed by aggregation of the amyloid beta-peptide (Abeta) are an intrinsic component of Alzheimer disease pathogenesis. It has been suggested that oxidation of methionine 35 in Abeta has implications for Alzheimer disease, and it has been shown that oxidation of Met-35 significantly inhibits aggregation in vitro. In this study, the aggregational properties of Abeta-(1-40) before and after Met-35 oxidation were investigated using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. The results show that Abeta (1-40)Met-35(O) trimer and tetramer formation is significantly attenuated as compared with Abeta-(1-40). This suggests that oxidation of Met-35 inhibits a conformational switch in Abeta-(1-40) necessary for trimer but not dimer formation. Random incorporation of Abeta-(1-40) and Abeta-(1-40)Met-35(O) in homo and heterooligomers could also be observed. This is the first report of an early rate-limiting step in Abeta-(1-40) aggregation. Slowing of the fibrillization process at this early step is likely to support prolonged solubility and clearance of Abeta from brain and may reduce disease progression. PMID- 11912199 TI - Accelerated plaque accumulation, associative learning deficits, and up-regulation of alpha 7 nicotinic receptor protein in transgenic mice co-expressing mutant human presenilin 1 and amyloid precursor proteins. AB - Familial Alzheimer's disease-associated mutations in presenilin 1 or 2 or amyloid precursor protein result in elevated beta-amyloid, beta-amyloid accumulation, and plaque formation in the brains of affected individuals. By crossing presenilin 1 transgenic mice carrying the A246E mutation with plaque-producing amyloid precursor protein K670N/M671L transgenic mice (Tg2576), we show that co expression of both mutant transgenes results in acceleration of amyloid accumulation and associative learning deficits. At 5 months of age with no detectable plaque pathology, amyloid precursor protein transgenic animals are impaired in contextual fear learning following two pairings of conditioned and unconditioned stimuli but appear normal following a more robust five-pairing training. At 9 months of age when beta-amyloid deposition is evident, these mice are impaired following both two-pairing and five-pairing protocols. Mice carrying both transgenes are impaired in contextual fear conditioning at either age. All transgenic animal groups performed as well as controls in cued fear conditioning, indicating that the contextual fear learning deficits are hippocampus-specific. The associative learning impairments are coincident with elevated alpha 7 nicotinic acetylcholine receptor protein in the dentate gyrus. These findings provide two robust and rapid assays for beta-amyloid-associated effects that can be performed on young animals: impaired contextual fear learning and up regulation of alpha 7 nicotinic receptors. PMID- 11912200 TI - Regulatory role for SRC and phosphatidylinositol 3-kinase in initiation of fibronectin matrix assembly. AB - Fibronectin (FN) matrix assembly is a tightly regulated stepwise process that is initiated by interactions between FN and cell surface integrin receptors. These interactions activate many intracellular signaling pathways that regulate processes such as cell adhesion, migration, and survival. Here we demonstrate that cells lacking Src family kinases showed reduced ability to assemble FN fibrils as detected by immunofluorescence and by analysis of detergent extracts. The amount of FN matrix was further reduced by treatment with the phosphatidylinositol 3 (PI 3-kinase) inhibitor, wortmannin. CHOalpha5 cells, which are dependent on exogenous FN to initiate fibril formation, also showed significant reductions in matrix when treated with inhibitors of Src and PI 3 kinase. Combination of both inhibitors showed an additive inhibitory effect on assembly, which was concomitant with a loss of focal adhesion kinase phosphorylation. Decreased binding of the 70-kDa amino-terminal FN fragment at matrix assembly sites further supports a role for these kinases early during the process. We propose that these two signaling molecules, which lie downstream of integrins and focal adhesion kinase, are essential for efficient initiation of FN matrix assembly. PMID- 11912201 TI - The endoplasmic reticulum-resident heat shock protein Gp96 activates dendritic cells via the Toll-like receptor 2/4 pathway. AB - The heat shock protein Gp96 has been shown to induce specific immune responses. On one hand, this phenomenon is based on the specific interaction with CD91 that mediates endocytosis and results in major histocompatibility complex class I restricted representation of the Gp96-associated peptides. On the other hand, Gp96 induces activation of professional antigen-presenting cells, resulting in the production of pro-inflammatory cytokines and up-regulation of costimulatory molecules by unknown mechanisms. In this study, we have analyzed the consequences of Gp96 interaction with cells expressing different Toll-like receptors (TLRs) and with bone marrow-derived dendritic cells from mice lacking functional TLR2 and/or TLR4 molecules. We find that the Gp96-TLR2/4 interaction results in activation of nuclear factor kappaB-driven reporter genes and mitogen- and stress activated protein kinases and induces IkappaBalpha degradation. Bone marrow derived dendritic cells of C3H/HeJ and more pronounced C3H/HeJ/TLR2(-/-) mice fail to respond to Gp96. Interestingly, activation of bone marrow-derived dendritic cells depends on endocytosis of Gp96 molecules. Our results provide, for the first time, the molecular basis for understanding the Gp96-mediated activation of antigen-presenting cells by describing the simultaneous stimulation of the innate and adaptive immune system. This feature explains the remarkable ability of Gp96 to induce specific immune responses against tumors and pathogens. PMID- 11912202 TI - A common mechanism of stage-regulated gene expression in Leishmania mediated by a conserved 3'-untranslated region element. AB - Developmental regulation of mRNA levels in trypanosomatid protozoa is determined post-transcriptionally and often involves sequences located in the 3' untranslated regions (3'-UTR) of the mRNAs. We have previously identified a developmentally regulated gene family in Leishmania encoding the amastin surface proteins and showed that stage-specific accumulation of the amastin mRNA is mediated by sequences within the 3'-UTR. Here we identified a 450-nt region within the amastin 3'-UTR that can confer amastigote-specific gene expression by a novel mechanism that increases mRNA translation without an increase in mRNA stability. Remarkably, this 450-nt 3'-UTR element is highly conserved among a large number of Leishmania mRNAs in several Leishmania species. Here we show that several of these mRNAs are differentially expressed in the intracellular amastigote stage of the parasite and that the 450-nt conserved element in their 3'-UTRs is responsible for stage-specific gene regulation. We propose that the 450-nt conserved element, which is unlike any other regulatory element identified thus far, is part of a common mechanism of stage-regulated gene expression in Leishmania that regulates mRNA translation in response to intracellular stresses. PMID- 11912203 TI - Hydroxylation and glycosylation of the four conserved lysine residues in the collagenous domain of adiponectin. Potential role in the modulation of its insulin-sensitizing activity. AB - It has recently been shown that the fat-derived hormone adiponectin has the ability to decrease hyperglycemia and to reverse insulin resistance. However, bacterially produced full-length adiponectin is functionally inactive. Here, we show that endogenous adiponectin secreted by adipocytes is post-translationally modified into eight different isoforms, as shown by two-dimensional gel electrophoresis. Carbohydrate detection revealed that six of the adiponectin isoforms are glycosylated. The glycosylation sites were mapped to several lysines (residues 68, 71, 80, and 104) located in the collagenous domain of adiponectin, each having the surrounding motif of GXKGE(D). These four lysines were found to be hydroxylated and subsequently glycosylated. The glycosides attached to each of these four hydroxylated lysines are possibly glucosylgalactosyl groups. Functional analysis revealed that full-length adiponectin produced by mammalian cells is much more potent than bacterially generated adiponectin in enhancing the ability of subphysiological concentrations of insulin to inhibit gluconeogenesis in primary rat hepatocytes, whereas this insulin-sensitizing ability was significantly attenuated when the four glycosylated lysines were substituted with arginines. These results indicate that full-length adiponectin produced by mammalian cells is functionally active as an insulin sensitizer and that hydroxylation and glycosylation of the four lysines in the collagenous domain might contribute to this activity. PMID- 11912204 TI - The role of region IVS5 of the human cardiac calcium channel in establishing inactivated channel conformation: use-dependent block by benzothiazepines. AB - The role of inactivated channel conformation and use dependence for diltiazem, a specific benzothiazepine calcium channel inhibitor, was studied in chimeric constructs and point mutants created in the IVS5 transmembrane segment of the L type cardiac calcium channel. All mutations, chimeric or point mutations, were restricted to IVS5, while the YAI-containing segment in IVS6, i.e. the primary interaction site with benzothiazepines, remained intact. Slowed inactivation rate and incomplete steady state inactivation, a behavior of some mutants, were accompanied by a reduced or by a complete loss of use-dependent block by diltiazem. Single channel properties of mutants that lost use dependence toward diltiazem were characterized by drastically elongated mean open times and distinctly slower time constants of open time distribution. Mutation of individual residues of the IVMLF segment in IVS5 did not mimic the complete loss of use dependence as observed for the replacement of the whole stretch. These results establish evidence that amino acids that govern inactivation and the drug binding site and other amino acids that are located distal from the putative drug binding site contribute significantly to the function of the benzothiazepine receptor region. The data are consistent with a complex "pocket" conformation that is responsive to a specific class of L-type calcium channel inhibitors. The data allow for a concept that multiple sites within regions of the alpha(1) subunit contribute to auto-regulation of the L-type Ca(2+) channel. PMID- 11912205 TI - Dynamics of gapped DNA recognition by human polymerase beta. AB - Kinetics of human polymerase beta binding to gapped DNA substrates having single stranded (ss) DNA gaps with five or two nucleotide residues in the ssDNA gap has been examined, using the fluorescence stopped-flow technique. The mechanism of the recognition does not depend on the length of the ssDNA gap. Formation of the enzyme complex with both DNA substrates occurs by a minimum three-step reaction, with the bimolecular step followed by two isomerization steps. The results indicate that the polymerase initiates the association with gapped DNA substrates through the DNA-binding subsite located on the 8-kDa domain of the enzyme. This first association step is independent of the length of the ssDNA gap and is characterized by similar rate constants for both examined DNA substrates. The subsequent, first-order transition occurs at the rate of approximately 600-1200 s(-1). This is the major docking step accompanied by favorable free energy changes in which the 31-kDa domain engages in interactions with the DNA. The 5' terminal PO(4)(-) group downstream from the primer is not a specific recognition element of the gap. However, the phosphate group affects the enzyme orientation in the complex with the DNA, particularly, for the substrate with a longer gap. PMID- 11912207 TI - Opposing roles for NF-kappa B/Rel factors p65 and c-Rel in the modulation of neuron survival elicited by glutamate and interleukin-1beta. AB - The nuclear transcription factors NF-kappaB/Rel have been shown to function as key regulators of either cell death or survival in neuronal cells. Here, we investigated whether selective activation of diverse NF-kappaB/Rel family members might lead to distinct effects on neuron viability. In both cultured rat cerebellar granule cells and mouse hippocampal slices, we examined NF-kappaB/Rel activation induced by two opposing modulators of cell viability: 1) interleukin 1beta (IL-1beta), which promotes neuron survival and 2) glutamate, which can elicit toxicity. IL-1beta produced a prolonged stimulation of NF-kappaB/Rel factors by inducing both IkappaBalpha and IkappaBbeta degradation. Glutamate produced a delayed and transient activation of NF-kappaB/Rel, which was associated with a brief loss of IkappaBalpha. Moreover, IL-1beta activated the p50, p65, and c-Rel subunits of NF-kappaB/Rel, whereas glutamate activated only the p50 and p65 proteins. The inhibition of NF-kappaB/Rel protein expression by antisense oligonucleotides in cerebellar granule cells showed that p65 was involved in glutamate-mediated cell death, whereas c-Rel was essential for IL 1beta-preserved cell survival. Furthermore, the depletion of c-Rel in cultured neurons as well as in the hippocampus from the c-Rel(-/-) mouse converted the IL 1beta effect into toxicity. These findings suggest that, within a single neuron, the balance between cell death and survival in response to external stimuli may rely on the activation of distinct NF-kappaB/Rel proteins. PMID- 11912206 TI - Role of tryptophan residues in interfacial binding of phosphatidylinositol specific phospholipase C. AB - The phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus thuringiensis exhibits several types of interfacial activation. In the crystal structure of the closely related Bacillus cereus PI-PLC, the rim of the active site is flanked by a short helix B and a loop that show an unusual clustering of hydrophobic amino acids. Two of the seven tryptophans in PI-PLC are among the exposed residues. To test the importance of these residues in substrate and activator binding, we prepared several mutants of Trp-47 (in helix B) and Trp-242 (in the loop). Two other tryptophans, Trp-178 and Trp-280, which are not near the rim, were mutated as controls. Kinetic (both phosphotransferase and cyclic phosphodiesterase activities), fluorescence, and vesicle binding analyses showed that both Trp-47 and Trp-242 residues are important for the enzyme to bind to interfaces, both activating zwitterionic and substrate anionic surfaces. Partitioning of the enzyme to vesicles is decreased more than 10-fold for either W47A or W242A, and removal of both tryptophans (W47A/W242A) yields enzyme with virtually no affinity for phospholipid surfaces. Replacement of either tryptophan with phenylalanine or isoleucine has moderate effects on enzyme affinity for surfaces but yields a fully active enzyme. These results are used to describe how the enzyme is activated by interfaces. PMID- 11912208 TI - Identification of epidermal growth factor receptor as a target of Cdc25A protein phosphatase. AB - Cdc25A, a dual-specificity protein phosphatase, plays a critical role in cell cycle progression. Although cyclin-dependent kinases are established substrates, Cdc25A may also affect other proteins. We have shown here that Cdc25A interacts with epidermal growth factor receptor (EGFR) both physically and functionally in Hep3B human hepatoma cells. Cdc25A inhibitor Cpd 5, a vitamin K analog, inhibited Cdc25A activity in the Cdc25A-EGFR immunocomplex and consequently caused prolonged EGFR tyrosine phosphorylation. Both purified GST-Cdc25A protein and endogenous Hep3B cellular Cdc25A dephosphorylated tyrosine-phosphorylated EGFR, and Cpd 5 antagonized the phosphatase activity of Cdc25A. A functional Cdc25A EGFR interaction was seen in NR-6 fibroblasts expressing ectopic EGFR but not with a receptor lacking the C terminus or a mutated kinase domain. These data link the cell cycle control Cdc25A phosphatase to an EGFR-linked mitogenic signaling pathway specifically involving EGFR dephosphorylation. PMID- 11912209 TI - Surfactant protein D gene regulation. Interactions among the conserved CCAAT/enhancer-binding protein elements. AB - Surfactant protein D (SP-D) plays roles in pulmonary host defense and surfactant homeostasis and is increased following acute lung injury. Given the importance of CCAAT/enhancer-binding protein (C/EBP)-binding elements in the systemic acute phase response and lung development and the expression of C/EBP isoforms by lung epithelial cells, we hypothesized that conserved C/EBP motifs in the near-distal and proximal promoters contribute to the regulation of SP-D expression by C/EBPs. Five SP-D motifs (-432, -340, -319, -140, and -90) homologous to the C/EBP consensus sequence specifically bound to C/EBPs in gel shift assays, and four of the five sites (-432, -340, -319, and -90) efficiently competed for the binding of C/EBPalpha, C/EBPbeta, or C/EBPdelta to consensus oligomers. Cotransfection of C/EBPalpha, C/EBPbeta, or C/EBPdelta cDNA in H441 lung adenocarcinoma cells significantly increased the luciferase activity of a wild-type SP-D promoter construct containing 698 bp of upstream sequence (SS698). Transfection of C/EBP also increased the level of endogenous SP-D mRNA in H441 cells. Transactivation of the reporter construct was abrogated by deletion of sequences upstream of 205. Independent site-directed mutagenesis of the sites at -432, -340, and -319 reduced C/EBP-mediated activation by approximately 50%, and mutagenesis of the site at -432 in combination with either of the tandem sites at -340 and -319 blocked activation. The conserved AP-1 element at -109 was required for maximal promoter activity, but not for the transactivation of SS698 by C/EBPs. Thus, interactions among C/EBP elements in the near-distal promoter can modulate the promoter activity of SP-D. PMID- 11912210 TI - Molecular determinants of human melanocortin-4 receptor responsible for antagonist SHU9119 selective activity. AB - The hypothalamic melanocortin-4 receptor (MC4R), a seven transmembrane G-protein coupled receptor, plays an important role in the regulation of body weight. The synthetic melanocortin analog SHU9119 has been widely used to characterize the physiological role of MC4R in feeding behavior and energy homeostasis. Previous studies indicated that SHU9119 is an agonist at the melanocortin-1 receptor (MC1R) but an antagonist at the MC4R. However, the molecular basis of the interaction between hMC4R and SHU9119 has not been clearly defined. To gain insight into the molecular determinants of hMC4R in the selectivity of SHU9119 chimeras and mutants hMC1R and hMC4R were expressed in cell lines and pharmacologically analyzed. A region of receptor containing the third transmembrane of hMC4R was found to be required for selective SHU9119 antagonism. Further mutagenesis studies of this region of hMC4R demonstrated that the amino acid residue leucine 133 in the third transmembrane was critical for the selective antagonist activity of SHU9119. The single substitution of leucine 133 to methionine did not affect SHU9119 binding to hMC4R. However, this substitution did convert SHU9119 from an antagonist to an agonist. Conversely, exchange of Met(128) in hMC1R to Leu, the homologous residue 133 of hMC4R, displayed a reduction in SHU9119 binding affinity and potency. This report provides the details of the molecular recognition of SHU9119 antagonism at hMC4R and shows that amino acid Leu(133) of hMC4R plays a key role in melanocortin receptor subtype specificity. PMID- 11912211 TI - Role of mammalian RAD51L2 (RAD51C) in recombination and genetic stability. AB - The highly conserved RAD51 protein has a central role in homologous recombination. Five novel RAD51-like genes have been identified in mammalian cells, but little is known about their functions. A DNA damage-sensitive hamster cell line, irs3, was found to have a mutation in the RAD51L2 gene and an undetectable level of RAD51L2 protein. Resistance of irs3 to DNA-damaging agents was significantly increased by expression of the human RAD51L2 gene, but not by other RAD51-like genes or RAD51 itself. Consistent with a role for RAD51L2 in homologous recombination, irs3 cells show a reduction in sister chromatid exchange, an increase in isochromatid breaks, and a decrease in damage-dependent RAD51 focus formation compared with wild type cells. As recently demonstrated for human cells, we show that RAD51L2 forms part of two separate complexes of hamster RAD51-like proteins. Strikingly, neither complex of RAD51-like proteins is formed in irs3 cells. Our results demonstrate that RAD51L2 has a key role in mammalian RAD51-dependent processes, contingent on the formation of protein complexes involved in homologous recombination repair. PMID- 11912213 TI - Activation of vascular endothelial growth factor A transcription in tumorigenic glioblastoma cell lines by an enhancer with cell type-specific DNase I accessibility. AB - Unregulated expression of vascular endothelial growth factor-A (VEGF-A) plays an important role in tumor growth. We have identified a cell type-specific enhancer, HS-1100, that contributes to VEGF-A transcriptional activation in tumorigenic glioblastoma cell lines. This enhancer exhibits increased accessibility to DNase I in glioblastoma cell lines that express high levels of VEGF-A but not in several other cell lines that express much lower levels of VEGF-A. HS-1100 contains a number of sequence elements that are highly conserved among human, mouse, and rat, including the hypoxia-response element (HRE). We show that the HRE contributes significantly to the cell type-specific enhancer activity of HS 1100 in U87MG glioblastoma cells. We use chromatin immunoprecipitation assays to show that endothelial PAS domain protein 1 (EPAS1) can efficiently bind to the endogenous HRE in U87MG cells but not in HEK293 cells in which the chromosomal HS 1100 is not accessible to DNase I. A dominant negative EPAS1 significantly reduces HS-1100 enhancer activity and VEGF-A levels in U87MG cells. Our results provide insight into the molecular mechanisms of VEGF-A up-regulation during cancer development. PMID- 11912214 TI - Endonuclease G, a candidate human enzyme for the initiation of genomic inversion in herpes simplex type 1 virus. AB - The herpes simplex virus type 1 (HSV-1) a sequence is present as a direct repeat at the two termini of the 152-kilobase viral genome and as an inverted repeat at the junction of the two unique components L and S. During replication, the HSV-1 genome undergoes inversion of L and S, producing an equimolar mixture of the four possible isomers. Isomerization is believed to result from recombination triggered by breakage at the a sequence, a recombinational hot spot. We have identified an enzyme in HeLa cell extracts that preferentially cleaves the a sequence and have purified it to near homogeneity. Microsequencing showed it to be human endonuclease G, an enzyme with a strong preference for G+C-rich sequences. Endonuclease G appears to be the only cellular enzyme that can specifically cleave the a sequence. Endonuclease G also showed the predicted recombination properties in an in vitro recombination assay. Based on these findings, we propose that endonuclease G initiates the a sequence-mediated inversion of the L and S components during HSV-1 DNA replication. PMID- 11912212 TI - Interaction of PIMT with transcriptional coactivators CBP, p300, and PBP differential role in transcriptional regulation. AB - PIMT (PRIP-interacting protein with methyltransferase domain), an RNA-binding protein with a methyltransferase domain capable of binding S-adenosylmethionine, has been shown previously to interact with nuclear receptor coactivator PRIP (peroxisome proliferator-activated receptor (PPAR)-interacting protein) and enhance its coactivator function. We now report that PIMT strongly interacts with transcriptional coactivators, CBP, p300, and PBP but not with SRC-1 and PGC 1alpha under in vitro and in vivo conditions. The PIMT binding sites on CBP and p300 are located in the cysteine-histidine-rich C/H1 and C/H3 domains, and the PIMT binding site on PBP is in the region encompassing amino acids 1101-1560. The N-terminal of PIMT (residues 1-369) containing the RNA binding domain interacts with both C/H1 and C/H3 domains of CBP and p300 and with the C-terminal portion of PBP that encompasses amino acids 1371-1560. The C-terminal of PIMT (residues 611-852), which binds S-adenosyl-l-methionine, interacts respectively with the C/H3 domain of CBP/p300 and with a region encompassing amino acids 1101-1370 of PBP. Immunoprecipitation data showed that PIMT forms a complex in vivo with CBP, p300, PBP, and PRIP. PIMT appeared to be co-localized in the nucleus with CBP, p300, and PBP. PIMT enhanced PBP-mediated transcriptional activity of the PPARgamma, as it did for PRIP, indicating synergism between PIMT and PBP. In contrast, PIMT functioned as a repressor of CBP/p300-mediated transactivation of PPARgamma. Based on these observations, we suggest that PIMT bridges the CBP/p300 anchored coactivator complex with the PBP-anchored coactivator complex but differentially modulates coactivator function such that inhibition of the CBP/p300 effect may be designed to enhance the activity of PBP and PRIP. PMID- 11912215 TI - Quaternary structure of coronavirus spikes in complex with carcinoembryonic antigen-related cell adhesion molecule cellular receptors. AB - Oligomeric spike (S) glycoproteins extend from coronavirus membranes. These integral membrane proteins assemble within the endoplasmic reticulum of infected cells and are subsequently endoproteolyzed in the Golgi, generating noncovalently associated S1 and S2 fragments. Once on the surface of infected cells and virions, peripheral S1 fragments bind carcinoembryonic antigen-related cell adhesion molecule (CEACAM) receptors, and this triggers membrane fusion reactions mediated by integral membrane S2 fragments. We focused on the quaternary structure of S and its interaction with CEACAMs. We discovered that soluble S1 fragments were dimers and that CEACAM binding was entirely dependent on this quaternary structure. However, two differentially tagged CEACAMs could not co precipitate with the S dimers, suggesting that binding sites were closely juxtaposed in the dimer (steric hindrance) or that a single CEACAM generated global conformational changes that precluded additional interactions (negative cooperativity). CEACAM binding did indeed alter S1 conformations, generating alternative disulfide linkages that were revealed on SDS gels. CEACAM binding also induced separation of S1 and S2. Differentially tagged S2 fragments that were free of S1 dimers were not co-precipitated, suggesting that S1 harbored the primary oligomerization determinants. We discuss the distinctions between the S.CEACAM interaction and other virus-receptor complexes involved in receptor triggered entry. PMID- 11912216 TI - Polycystin-1 activation of c-Jun N-terminal kinase and AP-1 is mediated by heterotrimeric G proteins. AB - Functional analysis of polycystin-1, the product of the gene most frequently mutated in autosomal dominant polycystic kidney disease, has revealed that this protein is involved in the regulation of diverse signaling pathways such as the activation of the transcription factor AP-1 and modulation of Wnt signaling. However, the initial steps involved in the activation of such cascades have remained unclear. We demonstrated previously that the C-terminal cytosolic tail of polycystin-1 binds and activates heterotrimeric G proteins in vitro. To test if polycystin-1 can activate cellular signaling cascades via heterotrimeric G protein subunits, polycystin-1 C-terminal tail-mediated c-Jun N-terminal kinase (JNK) and AP-1 activities were assayed in transiently transfected 293T cells in the presence of dominant-negative, G protein inhibiting constructs, and in the presence of cotransfected Galpha subunits. The results showed that polycystin-1 mediated JNK/AP-1 activation is mediated by Galpha and Gbetagamma subunits. Polycystin-1-mediated AP-1 activity could be significantly augmented by cotransfected Galpha(i), Galpha(q), and Galpha(12/13) subunits, suggesting that polycystin-1 can couple with and activate several heterotrimeric G protein families. PMID- 11912217 TI - Structure of formamidopyrimidine-DNA glycosylase covalently complexed to DNA. AB - Formamidopyrimidine-DNA glycosylase (Fpg) is a DNA repair enzyme that excises oxidized purines from damaged DNA. The Schiff base intermediate formed during this reaction between Escherichia coli Fpg and DNA was trapped by reduction with sodium borohydride, and the structure of the resulting covalently cross-linked complex was determined at a 2.1-A resolution. Fpg is a bilobal protein with a wide, positively charged DNA-binding groove. It possesses a conserved zinc finger and a helix-two turn-helix motif that participate in DNA binding. The absolutely conserved residues Lys-56, His-70, Asn-168, and Arg-258 form hydrogen bonds to the phosphodiester backbone of DNA, which is sharply kinked at the lesion site. Residues Met-73, Arg-109, and Phe-110 are inserted into the DNA helix, filling the void created by nucleotide eversion. A deep hydrophobic pocket in the active site is positioned to accommodate an everted base. Structural analysis of the Fpg DNA complex reveals essential features of damage recognition and the catalytic mechanism of Fpg. PMID- 11912218 TI - Neuroprotective effect of eicosapentaenoic acid in hippocampus of rats exposed to gamma-irradiation. AB - Exposure to irradiation leads to detrimental changes in several cell types. In this study we assessed the changes induced in hippocampus by exposure of rats to whole body irradiation; the findings reveal that irradiation leads to apoptotic cell death in hippocampus, and as a consequence, long term potentiation in perforant path-granule cell synapses is markedly impaired. The evidence is consistent with the view that irradiation induced an increase in reactive oxygen species and that this leads to stimulation of the stress-activated protein kinase, JNK, and activation of the transcription factor, c-Jun. Consequent upon activation of JNK, a cascade of cell signaling events was stimulated that ultimately resulted in apoptosis, as suggested by parallel increases in cytochrome c translocation, caspase-3 activation, poly(ADP-ribose) polymerase cleavage, and terminal dUTP nick-end labeling staining. Treatment of rats with eicosapentaenoic acid inhibited the irradiation-induced increase in reactive oxygen species production and the subsequent cellular signaling events, suggesting that oxidative stress triggered apoptotic cell death in the hippocampus of rats exposed to irradiation. Significantly, when the compromise in cell viability induced by irradiation was prevented by eicosapentaenoic acid, long term potentiation was sustained in a manner similar to that in the sham treated control group. PMID- 11912219 TI - Advanced glycation end product-induced apoptosis and overexpression of vascular endothelial growth factor and monocyte chemoattractant protein-1 in human cultured mesangial cells. AB - Advanced glycation end products (AGE) have been implicated in the pathogenesis of glomerulosclerosis in diabetes. However, their involvement in the development of the early phase of diabetic nephropathy has not been fully elucidated. We investigated the effects of AGE on growth and on vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1 (MCP-1) expression in human cultured mesangial cells. We prepared three immunochemically distinct AGE by incubating bovine serum albumin (BSA) with glucose, glyceraldehyde, or glycolaldehyde. When human mesangial cells were cultured with various types of AGE-BSA, viable cell numbers as well as DNA syntheses were significantly decreased. All of the AGE-BSA were found to significantly increase p53 and Bax protein accumulations and subsequently induce apoptotic cell death in mesangial cells. An antioxidant, N-acetylcysteine, significantly prevented the AGE-induced apoptotic cell death in mesangial cells. Human mesangial cells stimulated prostacyclin production by co-cultured glomerular endothelial cells. Furthermore, various types of AGE-BSA were found to up-regulate the levels of mRNAs for VEGF and stimulate the secretion of VEGF and MCP-1 proteins in mesangial cells. The results suggest that AGE disturbed glomerular homeostasis by inducing apoptotic cell death in mesangial cells and elicited hyperfiltration and microalbuminuria by stimulating the secretion of VEGF and MCP-1 proteins, thereby being involved in the pathogenesis of the early phase of diabetic nephropathy. PMID- 11912221 TI - Carbon and nitrogen assimilation in relation to yield: mechanisms are the key to understanding production systems. AB - Improved understanding of crop production systems in relation to N-supply has come from a knowledge of basic plant biochemistry and physiology. Gene expression leads to protein synthesis and the formation of metabolic systems; the ensuing metabolism determines the capacity for growth, development and yield production. This constitutes the genetic potential. These processes set the requirements for the supply of resources. The interactions between carbon dioxide (CO(2)) and nitrate () assimilation and their dynamics are of key importance for crop production. In particular, an adequate supply of, its assimilation to amino acids (for which photosynthesized carbon compounds are required) and their availability for protein synthesis, are essential for metabolism. An adequate supply of stimulates leaf growth and photosynthesis, the former via cell growth and division, the latter by larger contents of components of the light reactions, and those of CO(2) assimilation and related processes. If the supply of resources exceeds the demand set by the genetic potential then production is maximal, but if it is less then potential is not reached; matching resources to potential is the aim of agriculture. However, the connection between metabolism and yield is poorly quantified. Biochemical characteristics and simulation models must be better used and combined to improve fertilizer-N application, efficiency of N use, and yields. Increasing N-uptake at inadequate N-supply by increasing rooting volume and density is feasible, increasing affinity is less so. It would increase biomass and N/C ratio. With adequate N, at full genetic potential, more C assimilation per unit N would increase biomass, but energy would be limiting at full canopy. Increasing C-assimilation per unit N would increase biomass but decrease N/C at both large and small N-supply. Increasing production of all biochemical components would increase biomass and demand for N, and maintain N/C ratio. Changing C- or N-assimilation requires modifications to many processes to effect improvements in the whole system; genetic engineering/molecular biological alterations to single steps in the central metabolism are unlikely to achieve this, because targets are unclear, and also because of the complex interactions between processes and environment. Achievement of the long-term objectives of improving crop N-use and yield with fewer inputs and less pollution, by agronomy, breeding or genetic engineering, requires a better understanding of the whole system, from genes via metabolism to yield. PMID- 11912222 TI - N uptake and distribution in crops: an agronomical and ecophysiological perspective. AB - The rate of N uptake of crops is highly variable during crop development and between years and sites. However, under ample soil N availability, crop N accumulation is highly related to crop growth rate and to biomass accumulation. Critical N concentration has been defined as the minimum N concentration which allows maximum growth rate. Critical N concentration declines during crop growth. The relationship between critical N concentration and biomass accumulation over the growth period of a crop is broadly similar within major C(3) and C(4) cultivated species. Therefore, the critical N concentration concept is widely used in agronomy as the basis of the diagnosis of crop N status, and allows discrimination between situations of sub-optimal and supra-optimal N supply. The relationship between N and biomass accumulation in crops, relies on the interregulation of multiple crop physiological processes. Among these processes, N uptake, crop C assimilation and thus growth rate, and C and N allocation between organs and between plants, play a particular role. Under sub-optimal N supply, N uptake of the crop depends on soil mineral N availability and distribution, and on root distribution. Under ample N supply, N uptake largely depends on growth rate via internal plant regulation. Carbon assimilation of the crop is related to crop N through the distribution of N between mature leaves with consequences for leaf and canopy photosynthesis. However, although less commonly emphasized, carbon assimilation of the crop also depends on crop N through leaf area development. Therefore, crop growth rate fundamentally relies on the balance of N allocation between growing and mature leaves. Nitrogen uptake and distribution also depends on C allocation between organs and N composition of these organs. Within shoots, allocation of C to stems generally increases in relation to C allocation to the leaves over the crop growth period. Allocation of C and N between shoots and roots also changes to a large extent in relation to soil N and/or crop N. These alterations in C and N allocation between plant organs have implications, together with soil availability and carbon assimilation, on N uptake and distribution in crops. Therefore, N uptake and distribution in plants and crops involves many aspects of growth and development. Regulation of nitrogen assimilation needs to be considered in the context of these interregulatory processes. PMID- 11912224 TI - Integrated physiological and agronomic modelling of N capture and use within the plant. AB - Today farmers have several constraints to take into account in managing their crops: (i) competitiveness: productivity must be maintained or increased whereas inputs must be decreased, (ii) the environmental consequences of cultural practices: pesticide and fertilizer use must be decreased, and (iii) product quality must be improved and nitrogen nutrition is an important factor in harvest quality. These new constraints sometimes conflict: maximum yield is often obtained with large amounts of N, increasing the risks of N leaching. The determination of rates and dates for nitrogen application must become more precise in this context. Tools are required for the forecasting of crop requirements, the diagnosis of N deficiencies during the crop cycle and breeding of new adapted varieties. Models and diagnosis indicators have been developed to meet these needs, but those relating to nitrogen are often based on empirical relationships. Moreover, the available models and indicators often fail to account for cultivar-specific responses. The improvement of agronomic tools and the breeding of new varieties adapted to new cropping systems should be based on a thorough understanding of the key metabolic processes involved, and the relative contributions of these processes to yield determination in conditions of fluctuating N supply. For both purposes, more information is required about plant and crop N economy. In this paper, the way in which N absorption and use within the plant and crop, plant responses to deficiencies and excesses of nitrogen are taken into account in major agronomic models is described first. The level of sophistication of the modules comprising these models depends on operational objectives. Secondly, the ways in which the most recent molecular plant physiology findings can, and indeed should, be integrated into models at the crop and crop cycle levels are described. The potential value of this approach for improving current agronomic models and diagnostic tools, and for breeding more efficient varieties is also discussed. PMID- 11912225 TI - Nitrate transport in plants: which gene and which control? AB - Nitrate uptake by root cells is a key step of nitrogen metabolism and has been widely studied at the physiological level and, more recently, at the molecular level. Two classes of genes, NRT1 and NRT2, have been found to be potentially involved in the high and low affinity nitrate transport systems (HATS and LATS, respectively). The complexity of the molecular basis of nitrate uptake has been enhanced by the finding that in many plants both NRT1 and NRT2 classes are represented by multigene families. Furthermore, recent studies demonstrate that the control mechanisms that lead to an active protein at the plasma membrane act on gene transcription, modulating the steady-state levels of mRNA, and on the activation of the protein, possibly by a phosphorylation/dephosphorylation process. This is a review of recent progress in the characterization of the NRT2 nitrate transporters, the composition of this family in Arabidopsis, their possible role in nitrate acquisition, and some aspects of their regulation in plants. PMID- 11912223 TI - What stay-green mutants tell us about nitrogen remobilization in leaf senescence. AB - Leaf senescence has an important role in the plant's nitrogen economy. Chlorophyll catabolism is a visible symptom of protein mobilization. Genetic and environmental factors that interfere with yellowing tend to modify protein degradation as well. The chlorophyll-protein relationship is much closer for membrane proteins than it is for soluble or total leaf proteins. In stay-greens, genotypes with a specific defect in the chlorophyll catabolism pathway, soluble protein degradation during senescence may be close to normal, but light harvesting and reaction centre thylakoid membrane proteins are much more stable. Genes for the chlorophyll catabolism pathway and its control are important in the regulation of protein mobilization. Genes for three steps in the pathway are reported to have been isolated. The gene responsible for the stay-green phenotype in grasses and legumes has not yet been cloned but a fair amount is known about it. Pigment metabolism in senescing leaves of the Festuca-Lolium stay-green mutant is clearly disturbed and is consistent with a blockage at the ring-opening (PaO) step in chlorophyll breakdown. PaO is de novo synthesized in senescence and thought to be the key enzyme in the chlorophyll a catabolic pathway. The stay green mutation is likely to be located in the PaO gene, or a specific regulator of it. These genes may well be in the various senescence-enhanced cDNA collections that have been generated, but functional handles on them are currently lacking. When the stay-green locus from Festuca pratensis was introgressed into Lolium temulentum, a gene encoding F. pratensis UDPG pyrophosphorylase was shown to have been transferred on the same chromosome segment. A strategy is described for cloning the stay-green gene, based on subtractive PCR-based analyses of intergeneric introgressions and map-based cloning. PMID- 11912226 TI - The Arabidopsis dual-affinity nitrate transporter gene AtNRT1.1 (CHL1) is regulated by auxin in both shoots and roots. AB - The AtNRT1.1 (CHL1) gene of Arabidopsis encodes a dual-affinity nitrate transporter and contributes to both low and high affinity nitrate uptake. Localization studies have shown that CHL1 expression is preferentially targeted to nascent organs and growing regions of roots and shoots in Arabidopsis. In roots, CHL1 expression is concentrated in the tips of primary and lateral roots and is activated during lateral root initiation. In shoots, strong CHL1 expression is found in young leaves and developing flower buds. These findings suggest that CHL1 expression might be regulated by a growth signal such as the phytohormone auxin. To test this, auxin regulation of CHL1 was examined. Using transgenic Arabidopsis plants containing CHL1::GUS/GFP DNA constructs, it was found that treatment with exogenous auxin or introduction of the auxin overproducing mutations (yucca and rooty) resulted in a strong increase in CHL1::GUS/GFP signals in roots and leaves. When mature roots were treated with auxin to induce lateral root formation, CHL1::GFP signals were dramatically enhanced in dividing pericycle cells and throughout primordia development. RNA blot analysis showed that CHL1 mRNA levels in whole seedlings increase within 30 min of auxin treatment. The distribution of CHL1 expression in Arabidopsis roots and shoots was found to be similar to that of DR5::GUS, a synthetic, auxin responsive gene. These results indicate that auxin acts as an important signal regulating CHL1 expression and contributes to the targeting of CHL1 expression to nascent organs and root tips in Arabidopsis. PMID- 11912227 TI - Nitrite transport to the chloroplast in Chlamydomonas reinhardtii: molecular evidence for a regulated process. AB - Nitrite transport to the chloroplast is not a well documented process in spite of being a central step in the nitrate assimilation pathway. The lack of molecular evidence, as well as the easy diffusion of nitrite through biological membranes, have made this physiological process difficult to understand in plant nutrition. The aim of this review is to illustrate that nitrite transport to the chloroplast is a regulated step, intimately related to the efficiency of nitrate utilization. In Chlamydomonas reinhardtii, the Nar1;1 gene has been shown to have this role in nitrate assimilation. NAR1;1 corresponds to a plastidic membrane transporter protein related to the bacterial formate/nitrite transporters. At least four Nar1 genes might exist in Chlamydomonas. The existence of orthologous Nar1 genes in plants is discussed. PMID- 11912228 TI - The regulation of nitrate and ammonium transport systems in plants. AB - Inorganic nitrogen concentrations in soil solutions vary across several orders of magnitude among different soils and as a result of seasonal changes. In order to respond to this heterogeneity, plants have evolved mechanisms to regulate and influx. In addition, efflux analysis using (13)N has revealed that there is a co ordinated regulation of all component fluxes within the root, including biochemical fluxes. Physiological studies have demonstrated the presence of two high-affinity transporter systems (HATS) for and one HATS for in roots of higher plants. By contrast, in Arabidopsis thaliana there exist seven members of the NRT2 family encoding putative HATS for and five members of the AMT1 family encoding putative HATS for. The induction of high-affinity transport and Nrt2.1 and Nrt2.2 expression occur in response to the provision of, while down regulation of these genes appear to be due to the effects of glutamine. High affinity transport and AMT1.1 expression also appear to be subject to down regulation by glutamine. In addition, there is evidence that accumulated and may act post-transcriptionally on transporter function. The present challenge is to resolve the functions of all of these genes. In Aspergillus nidulans and Chlamydomonas reinhardtii there are but two high-affinity transporters and these appear to have undergone kinetic differentiation that permits a greater efficiency of absorption over the wide range of concentration normally found in nature. Such kinetic differentiation may also have occurred among higher plant transporters. The characterization of transporter function in higher plants is currently being inferred from patterns of gene expression in roots and shoots, as well as through studies of heterologous expression systems and knockout mutants. PMID- 11912229 TI - Interaction of cytosolic and plastidic nitrogen metabolism in plants. AB - In angiosperms, the assimilation of ammonia resulting from nitrate reduction and from photorespiration depends on the operation of the plastidic GS/GOGAT cycle. The precursor for ammonia assimilation, 2-oxoglutarate, is synthesized in the mitochondria and in the cytosol. It is imported into the plastid by a 2 oxoglutarate/malate translocator (DiT1). In turn, the product of ammonia assimilation, glutamate, is exported from the plastids by a glutamate/malate translocator (DiT2). These transport processes link plastidic and cytosolic nitrogen metabolism and are essential for plant metabolism. DiT1 was purified to homogeneity from spinach chloroplast envelope membranes and identified as a protein with an apparent molecular mass of 45 kDa. Peptide sequences were obtained from the protein and the corresponding cDNA was cloned. The function of the DiT1 protein and its substrate specificity were confirmed by expression of the cDNA in yeast cells and functional reconstitution of the recombinant protein into liposomes. Recent advances in the molecular cloning of DiT2 and in the analysis of the in vivo function of DiT1 by antisense repression in transgenic tobacco plants will be discussed. In non-green tissues, the reducing equivalents required for glutamate formation by NADH-GOGAT are supplied by the oxidative pentose phosphate pathway. Glucose 6-phosphate, the immediate precursor of the oxidative pentose phosphate pathway is generated in the cytosol and imported into the plastids by the plastidic glucose 6-phosphate/phosphate translocator. PMID- 11912230 TI - Modulation of nitrate reductase: some new insights, an unusual case and a potentially important side reaction. AB - The mechanism of the post-translational modulation of nitrate reductase activity (NR, EC 1.6.6.1) is briefly summarized, and it is shown that by this mechanism nitric oxide production through NR is also rapidly modulated. New and partly unexpected details on the modulation mechanism have been obtained by using immunological techniques. The phosphorylation state of NR has been assessed with peptide antibodies raised against the serine phosphorylation motive of spinach NR. By co-immunoprecipitation experiments, 14-3-3 binding to phospho-NR and the function of Mg(2+) in that process has been elucidated. Conflicting data on the role of NR phosphorylation and 14-3-3 binding in controlling NR proteolysis are discussed. A possible role of other NR inactivating proteins is also briefly considered and the regulation of NR of Ricinus communis is described as an interesting special case that differs from the 'normal' mechanism in several important aspects. PMID- 11912231 TI - The regulation of ammonium translocation in plants. AB - Much controversy exists about whether or not NH(+)(4) is translocated in the xylem from roots to shoots. In this paper it is shown that such translocation can indeed take place, but that interference from other metabolites such as amino acids and amines may give rise to large uncertainties about the magnitude of xylem NH(+)(4) concentrations. Elimination of interference requires sample stabilization by, for instance, formic acid or methanol. Subsequent quantification of NH(+)(4) should be done by the OPA-fluorometric method at neutral pH with 2-mercaptoethanol as the reducing agent since this method is sensitive and reliable. Colorimetric methods based on the Berthelot reaction should never be used, as they are prone to give erroneous results. Significant concentrations of NH(+)(4), exceeding 1 mM, were measured in both xylem sap and leaf apoplastic solution of oilseed rape and tomato plants growing with NO(-)(3) as the sole N source. When NO(-)(3) was replaced by NH(+)(4), xylem sap NH(+)(4) concentrations increased with increasing external concentrations and with time of exposure to NH(+)(4). Up to 11% of the translocated N was constituted by NH(+)(4). Glutamine synthetase (GS) incorporates NH(+)(4) into glutamine, but root GS activity and expression were repressed when high levels of NH(+)(4) were supplied. Ammonium concentrations measured in xylem sap sampled just above the stem base were highly correlated with NH(+)(4) concentrations in apoplastic solution from the leaves. Young leaves tended to have higher apoplastic NH(+)(4) concentrations than older non-senescing leaves. The flux of NH(+)(4) (concentration multiplied by transpirational water flow) increased with temperature despite a decline in xylem NH(+)(4) concentration. Retrieval of leaf apoplastic NH(+)(4) involves both high and low affinity transporters in the plasma membrane of mesophyll cells. Current knowledge about these transporters and their regulation is discussed. PMID- 11912232 TI - Molecular and enzymatic analysis of ammonium assimilation in woody plants. AB - Ammonium is assimilated into amino acids through the sequential action of glutamine synthetase (GS) and glutamate synthase (GOGAT) enzymes. This metabolic pathway is driven by energy, reducing power and requires the net supply of 2 oxoglutarate that can be provided by the reaction catalysed by isocitrate dehydrogenase (IDH). Most studies on the biochemistry and molecular biology of N assimilating enzymes have been carried out on annual plant species and the available information on woody models is far more limited. This is in spite of their economic and ecological importance and the fact that nitrogen is a common limiting factor for tree growth. GS, GOGAT and IDH enzymes have been purified from several woody species and their kinetic and molecular properties determined. A number of cDNA clones have also been isolated and characterized. Although the enzymes are remarkably well conserved along the evolutionary scale, major differences have been found in their compartmentation within the cell between angiosperms and conifers, suggesting possible adaptations to specific functional roles. The analysis of the gene expression patterns in a variety of biological situations such as changes in N nutrition, development, biotic or abiotic stresses and senescence, suggest that cytosolic GS plays a central and pivotal role in ammonium assimilation and metabolism in woody plants. The modification of N assimilation efficiency has been recently approached in trees by overexpression of a cytosolic pine GS in poplar. The results obtained, suggest that an increase in cytosolic GS might lead to a global effect on the synthesis of nitrogenous compounds in the leaves, with enhanced vegetative growth of transgenic trees. All these data suggest that manipulation of cytosolic GS may have consequences for plant growth and biomass production. PMID- 11912233 TI - Enzyme redundancy and the importance of 2-oxoglutarate in plant ammonium assimilation. AB - Ammonium is the reduced nitrogen form available to plants for assimilation into amino acids. This is achieved by the GS/GOGAT pathway that requires carbon skeletons in the form of 2-oxoglutarate. To date, the exact enzymatic origin of this organic acid for plant ammonium assimilation is unknown. Isocitrate dehydrogenases and aspartate aminotransferases have been proposed to carry out this function. Since different (iso)forms located in several subcellular compartments are present within a plant cell, recent efforts have concentrated on evaluating the involvement of these enzymes in ammonium assimilation. Furthermore, several observations indicate that 2-oxoglutarate is a good candidate as a metabolic signal to regulate the co-ordination of C and N metabolism. This will be discussed with respect to recent advances in bacterial signalling processes involving a 2-oxoglutarate binding protein called PII. PMID- 11912234 TI - Genetic manipulation and quantitative-trait loci mapping for nitrogen recycling in rice. AB - Immunocytological studies in this laboratory have suggested that NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) in developing organs of rice (Oryza sativa L. cv. Sasanishiki) is involved in the utilization of glutamine remobilized from senescing organs through the phloem. Because most of the indica cultivars contained less NADH-GOGAT in their sink organs than japonica cultivars, over-expression of NADH-GOGAT gene from japonica rice was investigated using Kasalath, an indica cultivar. Several T0 transgenic Kasalath lines over-producing NADH-GOGAT under the control of a NADH-GOGAT promoter of Sasanishiki, a japonica rice, showed an increase in grain weight (80% as a maximum), indicating that NADH GOGAT is indeed a key step for nitrogen utilization and grain filling in rice. A genetic approach using 98 backcross-inbred lines (BC(1)F(6)) developed between Nipponbare (a japonica rice) and Kasalath were employed to detect putative quantitative trait loci (QTLs) associated with the contents of cytosolic glutamine synthetase (GS1; EC 6.3.1.2), which is probably involved in the export of nitrogen from senescing organs and those of NADH-GOGAT. Immunoblotting analyses showed transgressive segregations toward lower or greater contents of these enzyme proteins in these BC(1)F(6). Seven chromosomal QTL regions were detected for GS1 protein content and six for NADH-GOGAT. Some of these QTLs were located in QTL regions for various biochemical and agronomic traits affected by nitrogen recycling. The relationships between the genetic variability of complex agronomic traits and traits for these two enzymes are discussed. PMID- 11912235 TI - Nitrogen metabolism and remobilization during senescence. AB - Senescence is a highly organized and well-regulated process. As much as 75% of total cellular nitrogen may be located in mesophyll chloroplasts of C(3)-plants. Proteolysis of chloroplast proteins begins in an early phase of senescence and the liberated amino acids can be exported to growing parts of the plant (e.g. maturing fruits). Rubisco and other stromal enzymes can be degraded in isolated chloroplasts, implying the involvement of plastidial peptide hydrolases. Whether or not ATP is required and if stromal proteins are modified (e.g. by reactive oxygen species) prior to their degradation are questions still under debate. Several proteins, in particular cysteine proteases, have been demonstrated to be specifically expressed during senescence. Their contribution to the general degradation of chloroplast proteins is unclear. The accumulation in intact cells of peptide fragments and inhibitor studies suggest that multiple degradation pathways may exist for stromal proteins and that vacuolar endopeptidases might also be involved under certain conditions. The breakdown of chlorophyll-binding proteins associated with the thylakoid membrane is less well investigated. The degradation of these proteins requires the simultaneous catabolism of chlorophylls. The breakdown of chlorophylls has been elucidated during the last decade. Interestingly, nitrogen present in chlorophyll is not exported from senescencing leaves, but remains within the cells in the form of linear tetrapyrrolic catabolites that accumulate in the vacuole. The degradation pathways for chlorophylls and chloroplast proteins are partially interconnected. PMID- 11912236 TI - Co-ordination of leaf minor amino acid contents in crop species: significance and interpretation. AB - The question of whether general control of amino acid synthesis exists in plants remains to be resolved. It is not known whether there is overall co-ordination of the biosynthesis of amino acids that are formed through distinct pathways. In this work, amino acid contents were measured in a large number of samples taken from wheat, potato and barley leaves under different photosynthetic conditions. The variability in total soluble amino acid contents between samples was approximately 6-fold in wheat and potato. Subtracting the major amino acids from the total soluble amino acids showed that the variability in summed minor amino acid contents was approximately 20-fold. This variability was not correlated with short-term changes in primary carbon and nitrogen metabolism, and only poorly correlated with total leaf amino acids. By contrast, striking linear relationships between the contents of most minor amino acids were observed, demonstrating that the contents of many minor amino acids vary in concert. These observations show that amino acid contents are co-ordinated across biosynthetic families. While these data might be interpreted as an indication of cross-pathway regulation of the expression of key biosynthetic enzymes, the impact of factors such as protein degradation and storage cannot be ignored. PMID- 11912237 TI - Cereal seed storage proteins: structures, properties and role in grain utilization. AB - Storage proteins account for about 50% of the total protein in mature cereal grains and have important impacts on their nutritional quality for humans and livestock and on their functional properties in food processing. Current knowledge of the structures and properties of the prolamin and globulin storage proteins of cereals and their mechanisms of synthesis, trafficking and deposition in the developing grain is briefly reviewed here. The role of the gluten proteins of wheat in determining the quality of the grain for breadmaking and how their amount and composition can be manipulated leading to changes in dough mixing properties is also discussed. PMID- 11912238 TI - Steps towards an integrated view of nitrogen metabolism. AB - This article discusses how nitrate assimilation is integrated with nitrate uptake, with ammonium assimilation and amino acid synthesis, with pH regulation, and with the sugar supply in tobacco leaves. During the first part of the light period, nitrate assimilation exceeds nitrate uptake by 2-fold and ammonium assimilation by 50%, leading to rapid depletion of nitrate and accumulation of ammonium, glutamine, glycine and serine. NIA, NII and PPC expression show a shared maximum early in the diurnal cycle to direct carbon towards malate synthesis for pH regulation. Later in the diurnal cycle an orchestrated increase of GLN2, PKc, CS, and ICDH-1 expression re-establishes a balance between nitrate assimilation and ammonium metabolism. Nitrate uptake continues throughout the night, replenishing the leaf nitrate pool. These diurnal changes are attenuated or abolished in mutants with low NIA activity, and modified in wild-type plants growing on different nitrogen sources or elevated [CO(2)]. Comparison across genotypes and conditions reveals that NIA transcript levels are always closely related to the balance between nitrate influx and assimilation, but are unrelated to changes of glutamine or 2-oxoglutarate. In a systematic search for other downstream regulators, a wide range of downstream metabolites was fed to detached leaves and glutamate, cysteine, asparagine, and malate identified as candidates. Low sugars totally inhibit nitrate assimilation, overriding signals from nitrogen metabolism. Moderate changes act post-transcriptionally, and larger changes lead to a collapse of the NIA transcript. Low sugars also lead to a collapse of minor amino acids and a dramatic decrease of phenylpropanoids and nicotine. Consequently, wild-type plants growing in unfavourable light regimes and antisense RBCS transformants are simultaneously carbon- and nitrogen-limited. PMID- 11912239 TI - Multiple routes communicating nitrogen availability from roots to shoots: a signal transduction pathway mediated by cytokinin. AB - In higher plants, inorganic nitrogen has crucial effects on growth and development, providing cellular components and modulating gene expression. To date, not only nitrogen assimilatory genes but also a substantial number of genes with other functions have been shown to be selectively regulated by the availability of nitrogen. In terms of the communicating substance(s) between root and shoot, accumulating evidence suggests that nitrate itself is the primary signal molecule triggering the activation of transcription of nitrate assimilation and related genes. On the other hand, some of the genes involved in photosynthesis, cell cycling and translation machinery are also regulated, at least in part, by nitrate and other nitrogen sources and, in some cases, the effect can be mimicked by cytokinin treatment. Spatial and temporal studies on the accumulation levels and the translocation of cytokinin in response to nitrate replenishment in maize showed subsequent accumulation of various cytokinin species in the roots, xylem sap and leaves. In Arabidopsis thaliana, trans-zeatin riboside-5'-monophosphate and/or trans-zeatin riboside also accumulated in the roots in response to nitrate resupply. These studies suggest that cytokinin metabolism and translocation could be commonly modulated by nitrogen availability in higher plants. Thus, in addition to nitrate, cytokinin could be another root to-shoot signal communicating nitrogen availability. PMID- 11912240 TI - The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. AB - This short review outlines the central role of glutamine synthetase (GS) in plant nitrogen metabolism and discusses some possibilities for crop improvement. GS functions as the major assimilatory enzyme for ammonia produced from N fixation, and nitrate or ammonia nutrition. It also reassimilates ammonia released as a result of photorespiration and the breakdown of proteins and nitrogen transport compounds. GS is distributed in different subcellular locations (chloroplast and cytoplasm) and in different tissues and organs. This distribution probably changes as a function of the development of the tissue, for example, GS1 appears to play a key role in leaf senescence. The enzyme is the product of multiple genes with complex promoters that ensure the expression of the genes in an organ- and tissue-specific manner and in response to a number of environmental variables affecting the nutritional status of the cell. GS activity is also regulated post translationally in a manner that involves 14-3-3 proteins and phosphorylation. GS and plant nitrogen metabolism is best viewed as a complex matrix continually changing during the development cycle of plants. Along with GS, a number of other enzymes play key roles in maintaining the balance of carbon and nitrogen. It is proposed that one of these is glutamate dehydrogenase (GDH). There is considerable evidence for a GDH shunt to return the carbon in amino acids back into reactions of carbon metabolism and the tri-carboxylic acid cycle. Results with transgenic plants containing transferred GS genes suggest that there may be ways in which it is possible to improve the efficiency with which crop plants use nitrogen. Marker-assisted breeding may also bring about such improvements. PMID- 11912241 TI - Contribution of the Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) to transepithelial transport of H(+), NH(4)(+), K(+), and Na(+) in rat outer medullary collecting duct. AB - In rat kidney, the "secretory" isoform of the Na-K-Cl cotransporter, NKCC1 (BSC 2), localizes to the basolateral membrane of the alpha intercalated cell, the acid secreting cell of the outer medullary collecting duct (OMCD). This laboratory has reported that NKCC1 mediates Cl(-) uptake across the basolateral membrane in series with Cl(-) secretion across the apical membrane in rat OMCD. NKCC1 transports NH(4)(+), K(+), and Na(+) as well as Cl(-); therefore, a role for the cotransporter in the process of HCl, NH(4)Cl, KCl, and NaCl secretion has been suggested. Thus, it was determined if bumetanide, an inhibitor of NKCC1, alters transepithelial cation transport in rat OMCD. OMCD tubules from deoxycorticosterone pivalate (DOCP)-treated rats were perfused in vitro. Hydration of CO(2), rather than NH(4)(+), provides the principle source of H(+) for net acid secretion. In HCO(3)(-)/CO(2)-buffered solutions, no effect of bumetanide on net K(+) flux was detected. Under some conditions, bumetanide addition resulted in a small reduction in secretion of net H(+) equivalents. Transepithelial Na(+) flux, J(Na), was -1.5 +/- 1.7 pmol/mm per min, values not different from zero. However, with the application of bumetanide to the bath, J(Na) was +5.2 +/- 1.3 pmol/mm per min (P < 0.05), which indicates net Na(+) absorption. In conclusion, inhibition of NKCC1 in rat OMCD changes transepithelial movement of Na(+) and Cl(-). The role of NKCC1 in the secretion of net H(+) equivalents is small. PMID- 11912242 TI - Human cortical distal nephron: distribution of electrolyte and water transport pathways. AB - The exact distributions of the different salt transport systems along the human cortical distal nephron are unknown. Immunohistochemistry was performed on serial cryostat sections of healthy parts of tumor nephrectomized human kidneys to study the distributions in the distal convolution of the thiazide-sensitive Na-Cl cotransporter (NCC), the beta subunit of the amiloride-sensitive epithelial Na channel (ENaC), the vasopressin-sensitive water channel aquaporin 2 (AQP2), and aquaporin 3 (AQP3), the H(+) ATPase, the Na-Ca exchanger (NCX), plasma membrane calcium-ATPase, and calbindin-D28k (CaBP). The entire human distal convolution and the cortical collecting duct (CCD) display calbindin-D28k, although in variable amounts. Approximately 30% of the distal convolution profiles reveal NCC, characterizing the distal convoluted tubule. NCC overlaps with ENaC in a short portion at the end of the distal convoluted tubule. ENaC is displayed all along the connecting tubule (70% of the distal convolution) and the CCD. The major part of the connecting tubule and the CCD coexpress aquaporin 2 with ENaC. Intercalated cells, undetected in the first 20% of the distal convolution, were interspersed among the segment-specific cells of the remainder of the distal convolution, and of the CCD. The basolateral calcium extruding proteins, Na-Ca exchanger (NCX), and the plasma membrane Ca(2+)-ATPase were found all along the distal convolution, and, in contrast to other species, along the CCD, although in varying amounts. The knowledge regarding the precise distribution patterns of transport proteins in the human distal nephron and the knowledge regarding the differences from that in laboratory animals may be helpful for diagnostic purposes and may also help refine the therapeutic management of electrolyte disorders. PMID- 11912243 TI - Immunolocalization of multispecific organic anion transporters, OAT1, OAT2, and OAT3, in rat kidney. AB - Recently, a family of multispecific organic anion transporters has been identified, and several isoforms have been reported. However, the physiologic and pharmacologic roles of each isoform, except OAT1, in the transepithelial transport of organic anions in the kidney remain to be elucidated. To address this issue, it is essential to determine the intrarenal distribution and membrane localization of each OAT isoform along the nephron. In this study, the intrarenal distributions of rOAT1, rOAT2, and rOAT3 were investigated by an immunofluorescence method that used frozen rat serial kidney sections. Confocal microscopic analysis showed that immunoreactivity for rOAT1 was detected exclusively in the proximal tubules (S1, S2, S3) in the cortex with basolateral membrane staining. rOAT2 was detected in the apical surface of the tubules in the medullary thick ascending limb of Henle's loop (MTAL) and cortical and medullary collecting ducts (CD). rOAT3 was localized in the basolateral digitation of the cell membrane in all the segments (S1, S2, and S3) of the proximal tubules, MTAL, cortical TAL, connecting tubules, and cortical and medullary CD. These results on the distribution of each OAT isoform will facilitate the understanding of the role of OATs in the renal processing of organic anions. PMID- 11912244 TI - Cisplatin induces apoptosis in LLC-PK1 cells via activation of mitochondrial pathways. AB - Cisplatin, a commonly used chemotherapeutic agent, has a major limitation because of its nephrotoxicity. Recent studies have shown that cisplatin causes apoptotic cell death in renal tubule cells, but the underlying molecular mechanisms remain to be elucidated. In this study, cisplatin was found to induce apoptosis in a dose- and duration-dependent manner in cultured proximal tubule (LLC-PK1) cells, as evidenced by DNA laddering and TdT-mediated dUTP nick end-labeling assay. Pretreatment with the specific caspase 9 inhibitor LEHD-CHO completely prevented the apoptosis, whereas the caspase 8 inhibitor IETD-fmk had no effect. Furthermore, the activity of caspase 9 was upregulated about sixfold by cisplatin in a dose-dependent manner. These results implicated the caspase 9-dependent mitochondrial apoptotic pathways. Indeed, cisplatin triggered a duration dependent translocation of cytochrome c from the mitochondria to the cytosol, by immunofluorescence and Western blots. Cisplatin treatment also resulted in the duration-dependent activation and mitochondrial translocation of the pro apoptotic molecule Bax, by immunofluorescence. Finally, cisplatin induced a duration-dependent onset of the mitochondrial permeability transition. Our results indicate that cisplatin induces apoptosis in LLC-PK1 cells via activation of mitochondrial signaling pathways. The sequence of events may be summarized as follows: activation of Bax induces mitochondrial permeability transition, leading to release of cytochrome c, activation of caspase 9, and entry into the execution phase of apoptosis. Inhibition of this specific pathway may provide a strategy to minimize cisplatin-induced nephrotoxicity. PMID- 11912245 TI - Gene expression levels and immunolocalization of organic ion transporters in the human kidney. AB - Renal excretion of organic anions and cations is mediated by the organic ion transporter family (SLC22A). In this study, the mRNA levels of the organic ion transporters were quantified by real-time PCR in normal parts of renal tissues from seven nephrectomized patients with renal cell carcinoma, and the distributions and localization of human (h)OAT1, hOAT3, and hOCT2 proteins were investigated by immunohistochemical analyses in the human kidney. The expression level of hOAT3 mRNA was the highest among the organic ion transporter family, followed by that of hOAT1 mRNA. The hOCT2 mRNA level was the highest in the human OCT family, and the level of hOCTN2 mRNA was higher than that of hOCTN1. hOCT1 mRNA showed the lowest level of expression in organic ion transporter family. hOAT1, hOAT3, and hOCT2 proteins were detected in crude membranes from the kidney of all patients by Western blot analyses, whereas hOCT1 protein could not be detected. Immunohistochemical analyses showed that both hOAT1 and hOAT3 were localized to the basolateral membrane of the proximal tubules in the cortex, and hOCT2 was localized to the basolateral membrane of the proximal tubules in both the cortex and medullary ray. Immunohistochemical analyses of serial sections indicated that hOAT1, hOAT3, and hOCT2 were coexpressed in a portion of the proximal tubules. These results suggest that hOAT1, hOAT3, and hOCT2 play predominant roles in the transport of organic ions across the basolateral membrane of human proximal tubules. PMID- 11912246 TI - Differential expression patterns of claudins, tight junction membrane proteins, in mouse nephron segments. AB - As the first step in understanding the physiologic functions of claudins (tight junction integral membrane proteins) in nephrons, the expression of claudin-1 to 16 in mouse kidneys was examined by Northern blotting. Among these claudins, only claudin-6, -9, -13, and -14 were not detectable. Claudin-5 and -15 were detected only in endothelial cells. Polyclonal antibodies specific for claudin-7 and -12 were not available. Therefore, the distributions of claudin-1, -2, -3, -4, -8, 10, -11, and -16 in nephron segments were examined with immunofluorescence microscopy. For identification of individual segments, antibodies specific for segment markers were used. Immunofluorescence microscopic analyses of serial frozen sections of mouse kidneys with polyclonal antibodies for claudins and segment markers revealed that claudins demonstrated very complicated, segment specific, expression patterns in nephrons, i.e., claudin-1 and -2 in Bowman's capsule, claudin-2, -10, and -11 in the proximal tubule, claudin-2 in the thin descending limb of Henle, claudin-3, -4, and -8 in the thin ascending limb of Henle, claudin-3, -10, -11, and -16 in the thick ascending limb of Henle, claudin 3 and -8 in the distal tubule, and claudin-3, -4, and -8 in the collecting duct. These segment-specific expression patterns of claudins are discussed, with special reference to the physiologic functions of tight junctions in nephrons. PMID- 11912247 TI - Blockade of the effects of TGF-beta1 on mesangial cells by overexpression of Smad7. AB - Smad7, a protein induced by transforming growth factor-beta1 (TGF-beta1) in many target cells, inhibits TGF-beta1 signal transduction and is thought to mediate an intracellular negative feedback response that limits TGF-beta1 effects. It is possible that overexpression of Smad7 could block specified effects of TGF-beta1 on mesangial cells, a TGF-beta target in glomerular disease. Smad7 mRNA was induced by TGF-beta1 within 1 h in a concentration-dependent manner in a transformed mouse mesangial cell (MMC) line. Uptake of (14)C-spermidine from the medium by MMC and the transcriptional activity of a segment of the human collagen pro-alpha2 type 1 chain (COL1A2) promoter fused to a luciferase reporter gene were used as indices of TGF-beta1. Treatment with TGF-beta1 increased (14)C spermidine uptake rate in a time-, concentration-, and temperature-dependent manner. For example, exposure to 1 ng/ml TGF-beta1 for 15 h increased uptake approximately twofold, a response that was attenuated by cycloheximide. Transfection of Smad7 expression vector into MMC abrogated both TGF-beta1 dependent stimulation of spermidine uptake and COL1A2 promoter activity. It is concluded that: (1) TGF-beta1 induces Smad7 in MMC; (2) (14)C-spermidine uptake is a convenient quantitative index of TGF-beta1 effect in these cells; and (3) overexpression of Smad7 is a highly effective method of blocking at least some mesangial cell effects of TGF-beta1 that may warrant evaluation in vivo in experimental glomerular disease. PMID- 11912248 TI - Role of high glucose-induced nuclear factor-kappaB activation in monocyte chemoattractant protein-1 expression by mesangial cells. AB - Although high glucose (HG) has been shown to induce nuclear factor-kappaB (NF kappaB) activation in vascular cells, the upstream regulation and the biologic significance of NF-kappaB activation in diabetic renal injury are not clear. It was, therefore, examined if HG-induced generation of reactive oxygen species (ROS) and protein kinase C (PKC) activation are involved in NF-kappaB activation in mesangial cells (MC), and the role of NF-kappaB activation in HG-induced monocyte chemoattractant protein-1 (MCP-1) expression by MC was further investigated. Recent observations suggest that MCP-1 may play a role in the development and progression of diabetic nephropathy. HG rapidly induced NF-kappaB activation in MC as estimated by electrophoretic mobility shift assay. Supershift assay suggests that most of the binding activity arose from p50/p50 and p50/p65 dimers. Antioxidants, pyrrolidine dithiocarbamate, N-acetyl-L-cystein, and trolox effectively inhibited HG-induced NF-kappaB activation in MC. HG rapidly generated dichlorofluorescin-sensitive intracellular ROS in MC as measured by laser scanning confocal microscopy. HG also activated PKC rapidly in MC. Inhibition of PKC effectively blocked HG-induced intracellular ROS generation and NF-kappaB activation in MC. HG increased MCP-1 mRNA expression by 1.9-fold and protein secretion by 1.6-fold that of control glucose in MC transfected with control vector but not in MC transfected with dominant negative mutant inhibitor of NF kappaB (IkappaBalphaM). Inhibition of either PKC or ROS effectively blocked HG induced, but not basal, MCP-1 protein secretion by MC transfected with control vector. Thus this study demonstrates that HG rapidly activates NF-kappaB in MC through PKC and ROS and suggests that HG-induced NF-kappaB activation in MC may play a role in diabetic renal injury through upregulation of MCP-1 mRNA and protein expression. PMID- 11912249 TI - Phosphatidylinositol 3-kinase activity is required for epidermal growth factor to suppress proteolysis. AB - Suppression of protein breakdown occurs commonly in cell growth, but the pathways responsible for controlling proteolysis are poorly understood. Protein breakdown in NRK-52E renal epithelial cells treated with epidermal growth factor (EGF) and intracellular signaling inhibitors or dominant negative signaling molecules contained in an adenoviral vector were measured. The tyrosine kinase inhibitor, herbimycin A, eliminated the suppression of proteolysis induced by EGF. In contrast, the Src inhibitor, PP1, had no effect. Expression of dominant negative H-RasY57 blocked the ability of EGF to stimulate downstream targets of Ras and also reduced the ability of EGF to suppress proteolysis. Inhibiting MEK did not influence the ability of EGF to suppress proteolysis, but the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor, LY249002, stimulated basal proteolysis and completely eliminated the proteolytic response to EGF. Use of an adenovirus that expresses a dominant negative p85 subunit of class 1 PI 3-kinase completely blocked the ability of EGF to suppress proteolysis, whereas use of an adenovirus expressing a K227E constitutively active p110 subunit reproduced the reduction in protein breakdown. It was concluded that EGF suppresses proteolysis by a mechanism that involves Ras and class 1 PI 3-kinase. PMID- 11912250 TI - Expression of a novel PDGF isoform, PDGF-C, in normal and diseased rat kidney. AB - Platelet-derived growth factor-C (PDGF-C) is a new member of the PDGF family. Its expression in normal and diseased kidney is unknown. Rabbit antisera were generated against human full-length, core domain, and mouse PDGF-C, and their specificity was confirmed by Western blot analyses. Renal PDGF-C expression was analyzed by immunohistochemistry in normal rats (n = 8), mesangioproliferative anti-Thy 1.1 nephritis (n = 4 each at days 1, 4, 6, and 85), passive Heymann nephritis (PHN, n = 4), puromycin nephrosis (PAN, n = 2), Milan normotensive rats (MN, n = 2), and obese Zucker rats (n = 3). PDGF-C expression was also studied in anti-Thy 1.1 rats treated with PDGF-B aptamer antagonists (n = 5) or irrelevant control aptamers (n = 5). PDGF-C was constitutively expressed in arterial smooth muscle cells and collecting duct epithelial cells. Mesangial PDGF-C was markedly upregulated in anti-Thy 1.1 nephritis in parallel with the peak mesangial cell proliferation. Furthermore, PDGF-CC acted as a potent growth factor for mesangial cells in vitro. Inhibition of PDGF-B via specific aptamers reduced the injury in anti-Thy 1.1 nephritis but did not affect the glomerular PDGF-C overexpression or the mitogenicity of PDGF-CC in vitro. In PHN, PAN, and obese Zucker rats, glomeruli remained negative for PDGF-C despite severe glomerular injury. PDGF-C localized to podocytes at sites of focal and segmental sclerosis in MN. Interstitial PDGF-C expression was increased at sites of fibrosing injury in obese Zucker rats. The use of the different antisera resulted in virtually identical findings. It is concluded that PDGF-C is a novel mesangial cell mitogen that is constitutively expressed in the kidney and specifically upregulated in mesangial, visceral epithelial, and interstitial cells after predominant injury to these cells. PDGF-C may therefore be involved in the pathogenesis of renal scarring. PMID- 11912251 TI - GAIP, GIPC and Galphai3 are concentrated in endocytic compartments of proximal tubule cells: putative role in regulating megalin's function. AB - Megalin is the most abundant endocytic receptor in the proximal tubule epithelium (PTE), where it is concentrated in clathrin-coated pits (CCPs) and vesicles in the brush border region. The heterotrimeric G protein alpha subunit, Galphai3, has also been localized to the brush border region of PTE. By immunofluorescence GIPC and GAIP, components of G protein-mediated signaling pathways, are also concentrated in the brush border region of PTE and are present in megalin expressing cell lines. By cell fractionation, these signaling molecules cosediment with megalin in brush border and microvillar fractions. GAIP is found by immunoelectron microscopy in CCPs, and GIPC is found in CCPs and apical tubules of endocytic compartments in the renal brush border. In precipitation assays, GST-GIPC specifically binds megalin. The concentration of Galphai3, GIPC, and GAIP with megalin in endocytic compartments of the proximal tubule, where extensive endocytosis occurs, and the interaction between GIPC and the cytoplasmic tail of megalin suggest a model whereby G protein-mediated signaling may regulate megalin's endocytic function and/or trafficking. PMID- 11912252 TI - C6 mediates chronic progression of tubulointerstitial damage in rats with remnant kidneys. AB - Although it was once considered only a marker of glomerular damage, accumulating evidence indicates that proteinuria per se is nephrotoxic and contributes to the progression of renal injury. Several studies have demonstrated that activation of complement in proteinuric urine results in tubular and interstitial damage. It was previously demonstrated that acute complement-mediated interstitial disease is induced by C5b-9. Here the role of C5b-9 in the progression of chronic proteinuric renal disease was investigated in a nonimmunologic remnant kidney model. Five-sixths nephrectomies were performed for normocomplementemic control and C6-deficient PVG rats. Tubulointerstitial injury was assessed by measurement of two independent markers of tubular injury (i.e., vimentin and osteopontin), interstitial accumulation of the extracellular matrix components collagen type I, collagen type IV, and laminin, interstitial macrophage infiltration, and renal function. The two groups developed similar levels of proteinuria and BP. Whereas C3 deposition on the brush border was equivalent for rats in the two groups, C5b 9 deposition was observed only for normocomplementemic rats. At day 35, the degrees of both tubulointerstitial injury and renal failure were the same for the two groups. Tubulointerstitial injury in normocomplementemic rats was still severe at day 70. In contrast, interstitial injury in C6-deficient rats had improved markedly at day 70, with improvements in renal function. In a rat model of chronic progressive renal disease secondary to nephron loss, the initial interstitial changes are complement-independent and largely reversible, whereas progressive interstitial fibrosis is mediated predominantly by C5b-9. Treatment to reduce C5b-9 attack in tubular cells may slow progression and facilitate recovery. PMID- 11912253 TI - Effects of mycophenolate mofetil in mercury-induced autoimmune nephritis. AB - Mycophenolate mofetil (MMF) is a new immunosuppressive drug whose active metabolite, mycophenolic acid (MPA), blocks the action of inosine monophosphate dehydrogenase, resulting in the inhibition of the novo purine synthesis. Thus, MPA has an antiproliferative effect on T and B lymphocytes and also inhibits the glycosylation of cell surface adhesion proteins involved in cell-cell contact and in the recruitment of circulating leukocytes to sites of tissue damage and inflammation. In this study, the effect of MMF in the mercury model of nephritis was examined. Repeated exposure to HgCl(2) induces an autoreactive Th2 cell subset-inducing polyclonal B cell activation in the Brown Norway (BN) rat. This leads to the development of an autoimmune syndrome characterized by synthesis of autoantibodies (mainly anti-glomerular basement membrane [GBM] Abs) with glomerular linear deposits of IgG, proteinuria, and tubulointerstitial nephritis. Results show that MMF has a preventive effect on mercury-induced disease as it blocks anti-GBM Ab synthesis, thus avoiding glomerular IgG deposits and proteinuria and the development of interstitial nephritis. However, the therapeutic effect of MMF seems to be restricted to its antiinflammatory properties blocking the extravasation of circulating leukocytes to renal interstitium by interfering with the very late activation antigen 4/vascular cell adhesion molecule-1 (VCAM-1) cell adhesion pathway. Also, MMF administration to mercury-injected rats reduces the secretion of the proinflammatory cytokine tumor necrosis factor-alpha. These findings confirm that MMF has a strong effect on the primary immune response in this model. Nevertheless, when the disease is in progress, MMF acts exclusively on the inflammatory response. MMF could be useful in the treatment of diseases associated with renal inflammation. PMID- 11912254 TI - Nephrin dissociates from actin, and its expression is reduced in early experimental membranous nephropathy. AB - These studies examined the expression of the podocyte slit diaphragm protein nephrin and its association with actin at the onset of proteinuria in passive Heymann nephritis (PHN), a rat model of human membranous nephropathy. Four days after immunization, 58% of PHN rats had mild proteinuria. At that time, most slit diaphragms were still visible on electron microscopy; however, in those locations where the deposits encroached on the filtration slits, the slit diaphragms were either displaced or absent. On day 7, the PHN rats were severely proteinuric, and most slit diaphragms were either absent, displaced, or replaced by occluding-type junctions. Immunofluorescence microscopy with antibodies to the external and cytoplasmic domains of nephrin showed a progressive loss of staining and a change in the distribution of nephrin from an interrupted linear pattern in normal controls to a more dispersed and clustered pattern in PHN. In contrast, the intensity of staining for ZO-1 and CD2-associated protein (CD2AP), two other proteins that are located on the cytoplasmic face of the slit diaphragm, was undiminished. Immunogold electron microscopy confirmed the progressive disappearance of nephrin from podocyte foot processes and retention of CD2AP. Glomeruli and glomerular cell membranes were extracted sequentially with Triton X 100, followed by DNase I or potassium iodide to depolymerize actin. Western blot analysis of the extracts showed a progressive decline of total nephrin on days 4 and 7 of PHN as well as a reduction in the actin-associated fraction. These findings show that nephrin partly dissociates from actin at the onset of podocyte injury in PHN. This is accompanied by a progressive loss of nephrin from the podocyte foot processes and prominent changes in the morphology of the slit diaphragms. These events may underlie the loss of podocyte barrier function in membranous nephropathy. PMID- 11912255 TI - Spatial and temporally restricted expression of chemokines and chemokine receptors in the developing human kidney. AB - The directed migration of cells, cell-cell adhesion, and the control of proliferation are key events during metanephric development. The chemokines are a family of proteins that selectively control aspects of cell migration, activation, proliferation, and adhesion. The expression of a series of chemokines and chemokine receptors during human renal development was investigated by using immunohistochemical analyses and real-time reverse transcription-PCR assays of defined laser-microdissected metanephric structures. The results demonstrate that mononuclear cell-like cells within the nephrogenic blastema focally express interferon-inducible protein-10/CXCL10, a ligand for CXCR3. Mononuclear-like cells dispersed through the developing organ express CX(3)CR1. Expression of CXCR4, the receptor for stromal cell-derived factor-1/CXCL12, is also limited to stromal CD34-positive cells. In contrast, the expression of stromal cell-derived factor-1/CXCL12, fractalkine, and CXCR3 is first observed in the comma- or S shaped body stage. The intensity of this expression becomes stronger in the capillary loop stage, and expression is mainly observed in the mesangial stalk and endothelial cells of the glomeruli. These proteins may play modulatory roles in kidney development. Because genes that are expressed during ontogeny often play a role in tissue regeneration, these embryonal chemokine/chemokine receptor patterns may be important in renal injury and repair. PMID- 11912256 TI - Loss-of-function polymorphism of the human kallikrein gene with reduced urinary kallikrein activity. AB - Kallikrein is synthesized in the distal tubules and produces kinins, which are involved in the regulation of vascular tone in the kidney. Urinary kallikrein activity has been reported to be partly inherited and to be reduced in essential hypertension. In a systematic search for molecular variants of the human kallikrein gene, nine single-nucleotide polymorphisms were identified. Five of those polymorphisms, including two nonsynonymous substitutions in exon 3, i.e., Arg53His (allelic frequency in Caucasian subjects, 0.03) and Gln121Glu (allelic frequency, 0.33), were studied in a normotensive group and two independent hypertensive groups for which 24-h urinary kallikrein activity had been measured. A significant decrease in urinary kallikrein activity was observed for the subjects who were heterozygous for the Arg53His polymorphism, compared with the other subjects. This finding was consistent in the two hypertensive groups and was observed with several kallikrein enzymatic assays. The Gln121Glu polymorphism and the other polymorphisms were not associated with changes in urinary kallikrein activity. None of the polymorphisms was associated with hypertension. Recombinant kallikrein variants were synthesized and enzymatically characterized, using native kininogen and kininogen-derived synthetic peptide substrates. No important effect was observed after Gln121 mutation, but there was a major decrease in enzyme activity when Arg53 was replaced by histidine. A model of kallikrein derived from crystallographic data suggested that Arg53 can affect substrate binding. The identification of a subset of subjects with genetically reduced kallikrein activity as a result of an amino acid mutation could facilitate analysis of the role of the kallikrein-kinin system in renal and vascular diseases. PMID- 11912257 TI - Engraftment and differentiation of human metanephroi into functional mature nephrons after transplantation into mice is accompanied by a profile of gene expression similar to normal human kidney development. AB - Metanephroi, the embryonic precursors of the adult kidney, can be induced in vivo to grow and develop. Despite their potential clinical utility for transplantation, the ability of human metanephroi to differentiate after transplantation into functional mature nephrons is mostly unknown. To address this, 70-d human metanephroi were transplanted into NOD/SCID mice; global gene expression patterns that underlie development of human metanephric transplants were analyzed and compared with normal human kidney development. In addition, functionality of the grafts was assessed by dimercaptosuccinic acid radioisotope scans at different times after transplantation. The results of hybridization to cDNA arrays when RNA was derived from normal human kidneys at 8, 12, 16, and 20 wk gestation demonstrated that a subset of 240 genes changed substantially with time. The induced genes can be classified as cell cycle regulators, transcription and growth factors, and signaling, transport, adhesion, and extracellular matrix molecules. Strikingly, clustering analysis of global gene expression at 2, 6, and 10 wk after metanephric transplantation revealed an expression profile that characterizes normal human kidney development. Moreover, maturation of the transplants was accompanied by an increased uptake of dimercaptosuccinic acid. Nevertheless, expression levels of specific genes were mostly found to be suppressed in the transplants compared with the normal kidneys. These data provide insights into human kidney development and support the possibility of the transplantability of human metanephroi. Understanding of the molecular regulation of the transplanted developing metanephroi might lead to the development of strategies aimed at increasing the levels of specific genes, nephron endowment, and graft function. PMID- 11912258 TI - Oxidant stress and reduced antioxidant enzyme protection in polycystic kidney disease. AB - Oxidative stress has been implicated in the pathogenesis of both acquired and hereditary polycystic kidney disease. Mechanisms of oxidant injury in C57BL/6J cpk mice and Han:SPRD-Cy rats with rapidly or slowly progressive polycystic kidney disease were explored. Expression of heme oxygenase-1 mRNA, an inducible marker of oxidative stress, was shown to be increased in cystic kidneys of mice and rats in a pattern that reflected disease severity. By contrast, there was a decrease in mRNA expression of the antioxidant enzymes extracellular glutathione peroxidase, superoxide dismutase, catalase, and glutathione S-transferase during disease progression. Renal mRNA levels of these enzymes were strikingly reduced in rapidly progressive disease in homozygous cystic mice and rats. In slowly progressive disease in heterozygous rats, renal antioxidant mRNA levels were decreased to a greater extent in cystic males than in the less severely affected females. Protein levels for extracellular glutathione peroxidase were also reduced in plasma and in cystic kidneys of mice and rats. Plasma extracellular glutathione peroxidase enzymatic activity was also decreased, whereas the lipid peroxidation products malondialdehyde and 4-hydroxy-2(E)-nonenal were increased in kidneys and blood plasma of cystic mice. Reduced antioxidant enzyme protection and increased oxidative damage represent general mechanisms in the pathogenesis of polycystic kidney disease. PMID- 11912259 TI - DNase I-like endonuclease in rat kidney cortex that is activated during ischemia/reperfusion injury. AB - Ischemia/reperfusion is known to result in DNA fragmentation and cell death in kidney tubular epithelium, but the endonucleases responsible for this DNA damage have not been identified. DNA substrate gel analysis of extracts from normal rat kidney cortex revealed the presence of a DNase with an apparent molecular mass of 30 to 34 kD. This enzyme is not a dimer of the previously described nuclear 15-kD endonuclease in kidney cells. Partially purified DNase exhibited characteristics similar to those of rat DNase I. The DNase was able to digest circular DNA (endonuclease), required both Ca(2+) and Mg(2+) ions, and was inhibited by Zn(2+) and by aurintricarboxylic acid; it was not inhibited by G-actin. Rat kidneys were subjected to 40 min of ischemia, followed by 0, 1, 4, 16, or 48 h of reperfusion. The activity of the DNase in cytosolic and nuclear extracts, the 200-bp ladder generating activity, and 3'OH strand breaks in nuclear DNA were simultaneously increased after ischemia, during the first hours of reperfusion. Oxidative DNA damage, measured as 8-hydroxydeoxyguanosine content, did not coincide with endonuclease-generated DNA breaks. Oxidative DNA damage was increased during ischemia and gradually decreased during reperfusion. Phosphorothioated DNase I antisense oligodeoxynucleotide introduced into cultured NRK-52E rat kidney epithelial cells inhibited DNA fragmentation and attenuated cell death induced by hypoxia/reoxygenation in vitro. The data indicate that the DNase I-like endonuclease may contribute to DNA fragmentation in reperfused rat kidneys. PMID- 11912260 TI - Impaired renal sensory responses after unilateral ureteral obstruction in the rat. AB - Renal responses to the activation of renal sensory receptors were examined in rats after release of 24-h unilateral ureteral obstruction of the left kidney. The integrity of the renorenal reflex was examined in both 24-h unilateral ureteral obstruction-treated (UUO) and sham-operated (Sham) rats. Increased ipsilateral afferent renal nerve activity (ARNA) and reflexly decreased efferent renal nerve activity (ERNA) and increased contralateral diuresis and natriuresis produced by increasing the left intrapelvic pressure were observed in Sham rats but not in UUO rats. The lack of responsiveness of the renorenal reflex in UUO rats was associated with lower release of substance P (SP) and increased neutral endopeptidase (NEP) activity in the renal pelvis in the postobstructive kidney. Compared with Sham rats, urine and sodium excretion after acute saline loading was significantly reduced in the postobstructive kidney. The blunted excretory responses were accompanied by lower activation of ARNA and less reflex inhibition of ERNA. Renal sensory dysfunction in the postobstructive kidney was further examined by stimulation of renal mechanoreceptors and chemoreceptors. Graded increases in intrapelvic pressure or renal pelvic perfusion with hypertonic saline solution elicited, respectively, a pressure- or concentration-dependent increase in ARNA in the control kidney of Sham rats, this response being greatly attenuated in the postobstructive kidney. Western blots showed no quantitative difference in the expression of renal pelvic neurokinin 1 (NK-1) receptors between the two groups. It was concluded that renal sensory function is impaired in the postobstructive kidney of UUO rats and that this defective activation of renal sensory receptors results in an impaired renorenal reflex, which is associated with enhanced NEP activity and catabolism of SP released in the renal pelvis and is not related to the expression of NK-1 receptor protein. PMID- 11912261 TI - The Calcimimetic agent AMG 073 lowers plasma parathyroid hormone levels in hemodialysis patients with secondary hyperparathyroidism. AB - Treatment with vitamin D sterols can lower plasma parathyroid hormone (PTH) in many patients with secondary hyperparathyroidism due to end-stage renal disease, but hypercalcemia, hyperphosphatemia, or both often develop during treatment. As such, alternative therapeutic approaches to managing excess PTH secretion are needed. Calcimimetic agents directly inhibit PTH secretion by activating the calcium-sensing receptor in the parathyroid glands, but clinical experience with them is limited. Fifty-two hemodialysis patients with secondary hyperparathyroidism were given single orally administered doses of the calcimimetic agent AMG 073 ranging from 5 to 100 mg, or placebo. Plasma PTH levels decreased 2 h after 25-, 50-, 75-, or 100-mg doses, falling by a maximum of 43 +/- 29%, 40 +/- 36%, 54 +/- 28%, or 55 +/- 39%, respectively. Plasma PTH levels decreased in all patients given doses of > or =25 mg but did not change in those who received placebo. In patients treated with daily doses of 25 or 50 mg of AMG 073 for 8 d, plasma PTH levels declined for the first 3 to 4 d and remained below baseline values after 8 d of treatment. Serum calcium concentrations also decreased by 5 to 10% from pretreatment levels in patients given 50 mg of AMG 073 for 8 d, but values were unchanged in those who received lower doses. Serum phosphorus levels and values for the calcium-phosphorus ion product both decreased after treatment with AMG 073. Thus, 8 d of treatment with AMG 073 effectively lowers plasma PTH levels and improves several disturbances in mineral metabolism that have been associated with soft tissue and vascular calcification and with adverse cardiovascular outcomes in patients with end-stage renal disease. PMID- 11912262 TI - Enhanced responses of blood pressure, renal function, and aldosterone to angiotensin I in the DD genotype are blunted by low sodium intake. AB - Angiotensin-converting enzyme (ACE) activity is increased in the DD genotype, but the functional significance for renal function is unknown. Blunted responses of BP and proteinuria to ACE inhibition among DD renal patients during periods of high sodium intake were reported. It was therefore hypothesized that sodium status affects the phenotype in the ACE I/D polymorphism. The effects of angiotensin I (AngI) and AngII among 27 healthy subjects, with both low (50 mmol sodium/d) and liberal (200 mmol sodium/d) sodium intakes, were studied. Baseline mean arterial pressure (MAP) values, renal hemodynamic parameters, and renin angiotensin system parameters were similar for all genotypes with either sodium intake level. With liberal sodium intake, the increases in MAP, renal vascular resistance, and aldosterone levels during AngI infusion (8 ng/kg per min) were significantly higher for the DD genotype, compared with the ID and II genotypes (all parameters presented as percent changes +/- 95% confidence intervals), with mean MAP increases of 22 +/- 2% (DD genotype), 13 +/- 5% (ID genotype), and 12 +/ 6% (II genotype) (P < 0.05), mean increases in renal vascular resistance of 100.1 +/- 19.7% (DD genotype), 73.0 +/- 16.3% (ID genotype), and 63.2 +/- 16.9% (II genotype) (P < 0.05), and increases in aldosterone levels of 650 +/- 189% (DD genotype), 343 +/- 71% (ID genotype), and 254 +/- 99% (II genotype) (P < 0.05). Also, the decrease in GFR was more pronounced for the DD genotype, with mean decreases of 17.9 +/- 4.7% (DD genotype), 8.8 +/- 3.4% (ID genotype), and 6.4 +/- 5.9% (II genotype) (P < 0.05). The effective renal plasma flow, plasma AngII concentration, and plasma renin activity values were similar for the genotypes. In contrast, with low sodium intake, the responses to AngI were similar for all genotypes. The responses to AngII were also similar for all genotypes, with either sodium intake level. In conclusion, the responses of MAP, renal hemodynamic parameters, and aldosterone concentrations to AngI are enhanced for the DD genotype with liberal but not low sodium intake. These results support the presence of gene-environment interactions between ACE genotypes and dietary sodium intake. PMID- 11912263 TI - Use of the albumin/creatinine ratio to detect microalbuminuria: implications of sex and race. AB - The recommended albumin (microg)/creatinine (mg) ratio (ACR) (30 microg/mg) to detect microalbuminuria does not account for sex or racial differences in creatinine excretion. In a nationally representative sample of subjects, the distribution of urine albumin and creatinine concentrations was examined by using one ACR value (> or =30 microg/mg) and sex-specific cutpoints (> or =17 microg/mg in men and > or =25 microg/mg in women) measured in spot urine specimens. Mean urine albumin concentrations were not significantly different between men and women, but urine creatinine concentrations were significantly higher (P < 0.0001). Compared with non-Hispanic whites, urine creatinine concentrations were significantly higher in non-Hispanic blacks (NHB) and Mexican Americans, whereas urine albumin concentrations were significantly higher in NHB (P < 0.0001) but not Mexican Americans. When a single ACR is used, the prevalence of microalbuminuria was significantly lower among the men compared with women (6.0 versus 9.2%; P < 0.0001) and among non-Hispanic whites compared with NHB (7.2 versus 10.2%; P < 0.0001). No significant difference in the prevalence of microalbuminuria between men and women was noted when sex-specific ACR cutpoints were used. In the multivariate adjusted model, female sex (odds ratio, 1.62; 95% confidence interval, 1.29 to 2.05) and NHB race/ethnicity (odds ratio, 1.34; 95% confidence interval, 1.12 to 1.61) were independently associated with microalbuminuria when a single ACR threshold was used. When a sex-specific ACR was used, NHB race/ethnicity remained significantly associated with microalbuminuria but sex did not. The use of one ACR value to define microalbuminuria may underestimate microalbuminuria in subjects with higher muscle mass (men) and possibly members of certain racial/ethnic groups. PMID- 11912264 TI - Feasibility of resuming peritoneal dialysis after severe peritonitis and Tenckhoff catheter removal. AB - Published guidelines suggest that after an episode of severe peritonitis that requires Tenckhoff catheter removal, peritoneal dialysis can be resumed after a minimum of 3 wk. However, the feasibility of resuming peritoneal dialysis after Tenckhoff catheter removal remains unknown. One hundred patients were identified with peritonitis that did not respond to standard antibiotic therapy in a specific center. Their clinical course was reviewed; in all of them, Tenckhoff catheters were removed and reinsertion was attempted at least 4 wk later. In 51 patients, the Tenckhoff catheter was successfully reinserted and peritoneal dialysis was resumed (success group). In the other 49 patients, reinsertion failed and the patient was put on long-term hemodialysis (fail group). The patients were followed for 18.5 +/- 16.8 mo. The overall technique survival was 30.8% at 24 mo. In the success group, 11 patients were changed to long-term hemodialysis within 8 mo after their return to continuous ambulatory peritoneal dialysis. In the fail group, 18 of the 20 deaths occurred within 12 mo after conversion to long-term hemodialysis. After resuming peritoneal dialysis, there was a significant decline in net ultrafiltration volume (0.38 +/- 0.16 to 0.21 +/ 0.19 L; P = 0.03) and a trend of rise in dialysate-to-plasma ratios of creatinine at 4 h (0.664 +/- 0.095 to 0.725 +/- 0.095; P = 0.15). Forty-five patients (88.2%) required additional dialysis exchanges or hypertonic dialysate to compensate for the loss of solute clearance or ultrafiltration, although there was no significant change in dialysis adequacy or nutritional status. It was concluded that after an episode of severe peritonitis that required Tenckhoff catheter removal, only a small group of patients could return to peritoneal dialysis. An early assessment of peritoneal function after Tenckhoff catheter reinsertion may be valuable. PMID- 11912265 TI - Postdialytic rebound of serum phosphorus: pathogenetic and clinical insights. AB - To gain insights into postdialytic rebound of serum phosphate (PDR-P), serum phosphate (P), calcium (Ca), and parathyroid hormone (PTH), levels were compared from the end of treatment (T0) to the subsequent 30 to 120 min and up to 68 hr in uremic patients who underwent with crossover modality a single session of two dialytic treatments characterized by different convective removal: standard hemodialysis (HD) and hemodiafiltration (HDF). In HDF, versus HD, P removal was greater (1171 +/- 90 versus 814 +/- 79 mg; P < 0.05) in the presence of similar predialytic P levels (6.0 +/- 0.2 and 5.9 +/- 0.4 mg/dl) and Kt/V (1.35 +/- 0.06 and 1.34 +/- 0.05); however, the serum P values at T0 did not differ (3.0 +/- 0.2 versus 3.3 +/- 0.2 mg/dl). In HDF, PDR-P was more rapid (30 min versus 90 min) and of a greater extent (at T120: +69 +/- 6% versus +31 +/- 4%; P < 0.0001). The higher P levels were maintained throughout the interdialytic period. Ca x P and PTH changed in parallel. Thereafter, patients were randomized to receive either HD or HDF for 3 mo. During this period, in the presence of similar Kt/V, protein intake, and dose of phospate binder, predialytic serum P levels diminished in HDF (from 5.8 +/- 0.2 to 4.4 +/- 0.3 mg/dl; P < 0.05), but they remained unchanged in HD. A similar pattern of changes was detected in Ca x P. Therefore, PDR-P is likely dependent on the mobilization of phosphate from a deep compartment induced by the intradialytic removal of this solute. Enhancement of convective removal acutely amplifies the entity of the phenomenon but allows a better control of Ca P homeostasis in the medium term. PMID- 11912266 TI - Peritoneal fluid and solute transport: influence of treatment time, peritoneal dialysis modality, and peritonitis incidence. AB - The integrity of the peritoneal membrane in peritoneal dialysis (PD) is of major importance for adequate dialysis and fluid balance. However, alterations in peritoneal fluid transport, such as ultrafiltration failure, often develop during long-term PD. To investigate peritoneal solute and fluid transport and to analyze the influence of treatment time, peritonitis incidence, and PD modality (continuous ambulatory PD [CAPD] or automated PD [APD]), a cross-sectional study with an extended peritoneal transport test that used dextran 70 in 2 L of glucose was performed in 23 nonselected chronic PD patients. Compared were long-term (>40 mo) with short-term PD patients (<40 mo), CAPD with APD patients, and those with a peritonitis incidence of >0.25/yr to those with an incidence of <0.25/yr. Dialysate/plasma (D/P) ratio and mass transfer area coefficient of creatinine, lymphatic absorption rate (LAR), transcapillary ultrafiltration, and effective ultrafiltration were measured. Long-term PD patients had higher D/P ratio of creatinine (73.5 +/- 2.3% versus 65.9 +/- 2.2%; P < 0.01) and higher LAR (243 +/- 69 ml/4 h versus 96 +/- 31 ml/4 h; P < 0.03), both resulting in lower effective ultrafiltration (242 +/- 35 ml/4 h versus 324 +/- 30 ml/4 h; P < 0.05). D/P ratio (r = 0.66) and LAR (r = 0.67) were positively correlated to PD duration. Patients on APD compared with those on CAPD and patients with a history of peritonitis compared with those without did not differ in terms of D/P ratio, mass transfer area coefficient, LAR, transcapillary ultrafiltration, and effective ultrafiltration. Lower ultrafiltration after long-term PD is both the result of increased small solute transport and increased lymphatic absorption. APD or CAPD modality and peritonitis incidence do not have a significant influence on small solute transport or fluid kinetics. PMID- 11912267 TI - Dialysis dose and body mass index are strongly associated with survival in hemodialysis patients. AB - Low dose of hemodialysis (HD) and small body size are independent risk factors for mortality. Recent changes in clinical practice, toward higher HD doses and use of more high-flux dialyzers, suggest the need to redetermine the dose level above which no benefit from higher dose can be observed. Data were analyzed from 45,967 HD patients starting end-stage renal disease (ESRD) therapy during April 1, 1997, through December 31, 1998. Data from Health Care Financing Administration (HCFA) billing records during months 10 to 15 of ESRD were used to classify each patient into one of five categories of HD dose by urea reduction ratio (URR) ranging from <60% to >75%. Cox regression models were used to calculate relative risk (RR) of mortality after adjustment for demographics, body mass index (BMI), and 18 comorbid conditions. Of the three body-size groups, the lowest BMI group had a 42% higher mortality risk than the highest BMI tertile. In each of three body-size groups by BMI, the RR was 17%, 17%, and 19% lower per 5% higher URR category among groups with small, medium, and large BMI, respectively (P < 0.0001 for each group). Patients treated with URR >75% had a substantially lower RR than patients treated with URR 70 to 75% (P < 0.005 each, for medium and small BMI groups). It is concluded that a higher dialysis dose, substantially above the Dialysis Outcomes Quality Initiative guidelines (URR >65%), is a strong predictor of lower patient mortality for patients in all body-size groups. Further reductions in mortality might be possible with increased HD dose. PMID- 11912268 TI - Matching older kidneys with older patients does not improve allograft survival. AB - Centers may restrict the use of some donor kidneys on the belief that overall graft survival is improved by giving older kidneys to older recipients and vice versa. The prevalence and the effect on graft survival (determined by death, return to dialysis, or retransplantation) of this practice among 74,297 first cadaver kidney transplantations in 1988 to 1998 was examined by using data from the United States Renal Data System. Giving older kidneys to older recipients is common; recipients > or =55 yr old received donor kidneys that were > or =55 yr old 46.2% more often than expected, but they received kidneys that were 18 to 29 yr old 33.6% less often than expected (chi(2) P < 0.0001). Both recipient and donor age have important effects on graft survival, although the effects of donor age are much stronger than those of recipient age. Compared with recipients 18 to 29 yr old, recipients > or =55 yr old were 25% (95% confidence interval, 15 to 35%, P < 0.0001) more likely to have graft failure (adjusted for donor age and other risk factors). On the other hand, donor kidneys > or =55 yr old were 78% (95% confidence interval, 58 to 99%, P < 0.0001) more likely to fail compared with kidneys 18 to 29 yr old. However, giving older kidneys to older recipients had little independent effect on graft survival, once the intrinsic effects of recipient and donor age were taken into account. For example, transplanting donor kidneys > or =55 yr old into recipients > or =55 yr old reduced the risk of graft failure only -6% (95% confidence interval, -18 to 8%, P = 0.3923) after the independent effects of donor and recipient age per se were taken into account. Thus, giving older kidneys to older recipients is a common practice that does not improve overall graft survival. PMID- 11912269 TI - First human trial of FTY720, a novel immunomodulator, in stable renal transplant patients. AB - FTY720 is a novel immunomodulator to be developed for use in organ transplantation. The primary objective of this study was to measure safety, single-dose pharmacokinetics, and pharmacodynamics in stable renal transplant patients-the first human use of FTY720. This study used a randomized, double blind, placebo-controlled design that explored single oral doses of FTY720 from 0.25 to 3.5 mg in 20 stable renal transplant patients on a cyclosporine-based regimen. Safety assessments and blood samples were taken predose and at multiple time points during a 96-h period postdose. Standard pharmacokinetic parameters were derived from the FTY720 whole blood concentrations, measured by HPLC/MS/MS. FTY720 was well tolerated, with no serious adverse events. Transient, asymptomatic bradycardia occurred after administration in 10 of 24 doses of FTY720. Pharmacokinetics are characterized by a prolonged absorption phase; the terminal elimination phase started 36 h after the administration, with elimination half-life (t(1/2)) ranging from 89 to 157 h independent of dose. Maximum plasma concentration and AUC were proportional to dose with low intersubject variability, the apparent volume of distribution (V(d)/F) ranged from 1116 to 1737 L. FTY pharmacodynamics were characterized by a reversible transient lymphopenia within 6 h, the nadir being 42% of baseline. The lymphocyte count returned to baseline within 72 h in all dosing cohorts except the highest. Single oral doses of FTY720 ranging from 0.25 to 3.5 mg were well tolerated and caused a reversible selective lymphopenia. Transient, but asymptomatic bradycardia was the most common adverse event. The long t(1/2) suggests less frequent dosing intervals. The size of V(d)/F is in excess of blood volume, consistent with widespread tissue distribution PMID- 11912270 TI - Congestive heart failure in renal transplant recipients: risk factors, outcomes, and relationship with ischemic heart disease. AB - Cardiovascular disease (CVD) is the major cause of death in renal transplant recipients (RTR). Several cohort studies have examined CVD in RTR, but none have addressed the development of congestive heart failure (CHF). CHF would hypothetically be a frequent and prognostically important event in RTR. A retrolective cohort study was, therefore, conducted in two Canadian centers to describe the incidence, risk factors for, and interrelationships between de novo CHF, de novo ischemic heart disease (IHD), and mortality in 638 consecutive adult RTR who were free of cardiac disease at 1 yr posttransplant. Detailed clinic and hospital records were available for 99% of patients. Median follow-up was 7 yr (range, 1 to 28 yr). De novo CHF occurred as frequently as de novo IHD (1.26 versus 1.22 events/100 patient-years, respectively) and appeared to carry a similar prognosis (relative risk for death, 1.78 [confidence interval, 1.21 to 2.61] for CHF versus 1.50 [1.05 to 2.13] for IHD). The incidence of CHF was considerably higher than that in the Framingham cohort, whereas the incidence of IHD was not, suggesting that renal transplantation might correspond more to a state of "accelerated heart failure" than to "accelerated atherosclerosis." Age, diabetes, gender, BP, and anemia were identified as independent risk factors for de novo CHF, whereas age, diabetes, gender, BP, and rejection were independent risk factors for de novo IHD. Optimal strategies for treatment of BP and anemia in RTR will need to be determined in randomized controlled trials. PMID- 11912271 TI - Capillary C4d deposition in kidney allografts: a specific marker of alloantibody dependent graft injury. AB - Capillary deposition of the complement split product C4d has been discussed as a marker for antibody-mediated kidney allograft rejection. The relationship between C4d staining and posttransplant alloantibody detection remains to be thoroughly investigated, however. In this study, C4d staining in peritubular capillaries (PTC) and the incidence of alloantibody formation, as detected with sensitive techniques, were evaluated among a cohort of transplant recipients who had undergone biopsies and had not been selected for a specific histologic diagnosis. One hundred thirteen biopsies, obtained from 58 cadaveric kidney transplant recipients, were tested. Serum samples obtained at the time of biopsy were evaluated by flow cytometric crossmatch (FCXM) testing and FlowPRA (One Lambda, Inc., Canoga Park, CA) analysis of anti-HLA panel reactivity. Most biopsies with C4d deposits in PTC (C4d(PTC)(+), n = 21 of 24) were associated with positive posttransplant FCXM results (T and/or B cell FCXM) and/or > or =5% FlowPRA (anti HLA class I and/or II) reactivity. Approximately 50% of the C4d(PTC)(-) biopsies were observed to be associated with donor-specific alloantibodies. Accordingly, high specificity (93%) but low sensitivity (31%) were calculated for capillary C4d staining (with FCXM testing as the standard method). For clinical evaluation, three patient groups were defined, i.e., a group of recipients with positive C4d staining in at least one allograft biopsy (C4d(PTC)(+), n = 16) and two C4d(PTC)( ) groups, which were discriminated on the basis of posttransplant FCXM results as C4d(PTC)(-)/FCXM(+) (n = 22) and C4d(PTC)(-)/FCXM(-) (n = 20) groups. Univariate analyses revealed significant differences between these groups with respect to serum creatinine levels at 12 mo [median, 2.83 mg/dl (interquartile range, 1.93 to 4.2 mg/dl) versus 1.78 mg/dl (1.47 to 2.24 mg/dl) versus 1.59 mg/dl (1.2 to 1.71 mg/dl), P < 0.001]. Of the five immunologic graft losses, four occurred in the C4d(PTC)(+) group and one occurred in the C4d(PTC)(-)/FCXM(+) group. In a multivariate analysis, C4d positivity was observed to have an independent predictive value for inferior 12-mo graft function (P = 0.02), whereas the observed moderate difference between C4d(PTC)(-)/FCXM(+) and C4d(PTC)(-)/FCXM(-) recipients did not achieve significance. In conclusion, these data demonstrate that positive C4d staining, which is an independent predictor of kidney graft dysfunction, represents a reliable specific marker for antibody-dependent graft injury. PMID- 11912273 TI - Pancreas after kidney transplantation. PMID- 11912272 TI - ACE inhibitors versus AT(1) receptor antagonists in patients with chronic renal disease. PMID- 11912274 TI - 2001 presidential address. PMID- 11912275 TI - Manipulating the calcium receptor. PMID- 11912276 TI - It's a Smad world: regulation of TGF-beta signaling in the kidney. PMID- 11912278 TI - Functional characterization of the human FSH receptor with an inactivating Ala189Val mutation. AB - An Ala189Val mutation of the human FSH receptor (FSHR) has been found to cause hypergonadotrophic ovarian failure with arrest of follicular maturation in women, and suppressed spermatogenesis in men. We have now characterized the molecular mechanisms of the receptor inactivation. Wild-type and mutant FSHR cDNAs were expressed in monkey kidney (COS-7) cells and murine granulosa tumour (KK-1) cells. Similar steady-state levels of FSHR mRNA were found in COS-7 and KK-1 cells transfected with both types of FSHR cDNA. Conspicuously, immunofluorescence and confocal microscopy studies revealed that whereas the wild-type receptor could be readily detected on the plasma membrane, most of the mutated protein was intracellularly sequestered. Ligand binding studies confirmed the greatly reduced cell surface expression of the mutant FSHR. A low level of mutated receptors were expressed at the cell surface, as shown by ligand binding and cAMP response. The capacity of these receptors to evoke another second messenger response, that of inositol trisphosphate (IP3), was almost totally lost. This finding may be related to the clinical picture of the patients, i.e. blockade of follicular maturation. There is a highly conserved stretch of five amino acids (Ala-Phe-Asn Gly-Thr) in the region of the mutation in all glycoprotein hormone receptors. We therefore created the same Ala to Val transition in the human LHR and studied its functional consequences. Similar functional alterations, i.e. intracellular sequestration and attenuated signal transduction, were found, as with mutated FSHR. Hence, this particular mutation in the conserved extracellular region of glycoprotein hormone receptors induces a conformational change that suppresses cell membrane targeting of the mutated receptor, probably through altered intracellular folding. PMID- 11912280 TI - Inhibitors of receptor tyrosine kinases do not suppress progesterone-induced [Ca2+]i signalling in human spermatozoa. AB - Previous studies have implicated receptor tyrosine kinases in progesterone induced [Ca2+]i signalling, and consequent induction of the acrosome reaction, in human spermatozoa. We have investigated the effects of tyrosine kinase inhibition on [Ca2+]i responses in large numbers of individual human spermatozoa. Genistein (5, 50 and 250 micromol/l), an inhibitor of receptor-linked tyrosine kinases, significantly inhibited the progesterone-induced acrosome reaction (P < 0.05). However, we could detect no effect of genistein on progesterone-induced [Ca2+]i signalling. In control experiments, application of progesterone induced a significant transient [Ca2+]i response in approximately 77% of cells and a sustained [Ca2+]i ramp/plateau in approximately 48% of cells (n = 26; 5411 cells). In preparations pretreated with genistein (50 micromol/l), significant transient and sustained responses were detected in 69.5 and 39.1% of cells respectively (n = 5; 1109 cells). The amplitudes of both transient and sustained [Ca2+]i responses were similar in control and genistein-pretreated preparations. Tyrphostin A47 (100 micromol/l), another receptor tyrosine kinase inhibitor, also failed to inhibit either the transient or sustained [Ca2+]i response (n = 3; 468 cells). Assessment of tyrosine phosphorylation of two sperm proteins (p105/81) showed greatly increased levels of phosphotyrosine in response to capacitation but a negligible increase in response to progesterone stimulation. Pretreatment with genistein (50 and 250 micromol/l) decreased capacitation-induced tyrosine phosphorylation and resulted in a loss of phosphorylation in response to progesterone treatment. We conclude that neither the transient nor sustained phases of the progesterone-induced [Ca2+]i response require receptor tyrosine kinase signalling. Previous reports of modulation of the progesterone-induced [Ca2+]i signal by tyrosine kinase inhibition probably reflect inhibition of the acrosome reaction. PMID- 11912279 TI - Transforming growth factor-beta1 inhibits steroidogenesis in human trophoblast cells. AB - Transforming growth factor-beta (TGF-beta) is an important regulator of placental development and function. In this study, we have investigated the effect of TGF beta1 on steroidogenesis, as well as its sites of action in the steroidogenic pathway by using a choriocarcinoma cell line, JEG-3, and a normal trophoblast cell line (NPC). The effect of TGF-beta1 on progesterone and estradiol production was evaluated in the absence or presence of a membrane-permeable analogue of cholesterol and some intermediate substrates of steroidogenic enzymes. The effect of TGF-beta1 on P450 aromatase (P450arom) mRNA levels was determined by Northern blot analysis. TGF-beta1 significantly decreased progesterone production in both NPC and JEG-3 cells. The inhibitory effect of TGF-beta1 on progesterone production was reversed by addition of 22R-hydroxycholesterol, a membrane permeable analogue of cholesterol, or pregnenolone. In JEG-3 cells, TGF-beta1 also inhibited estradiol production when androstenedione, but not estrone, was added to the culture. Estradiol production was too low to be detected in NPC cells. Treatment with TGF-beta1 also suppressed aromatase mRNA levels. This study has demonstrated that TGF-beta1 inhibits progesterone and estradiol production by trophoblast cells, and that the sites of TGF-beta1 action on progesterone and estradiol production are likely to be cholesterol transport and P450arom respectively. PMID- 11912281 TI - Effect of mifepristone and levonorgestrel on expression of steroid receptors in the human Fallopian tube. AB - It is likely that mifepristone or levonorgestrel in the future will find extended use for contraceptive purposes. It is therefore essential to characterize the modes of action of these compounds. To assess the effect on the human Fallopian tube, 24 women with regular menstrual cycles and proven fertility, admitted to the hospital for voluntary sterilization by laparoscopic technique, were randomly allocated to a control or one of two treatment groups. Treatments were given with either a single dose of 200 mg mifepristone or 0.75 mg levonorgestrel in two doses 12 h apart, on day LH+2. Surgery was performed on day LH+4 to LH+6. Steroid receptor expression was analysed by immunohistochemistry, Western blot and RT PCR. In the controls, there was a higher concentration of progesterone receptors in the stromal cells in the isthmic region than in those in the ampullar region. Treatment with mifepristone increased the progesterone receptor concentration in epithelial and stromal cells and increased the estrogen receptor concentration in epithelial cells. No effect on steroid receptor concentration was found following levonorgestrel. The contraceptive effect of post-ovulatory mifepristone has previously been considered to be dependent on an effect on the endometrium. However an effect on the Fallopian tube could contribute to alter the peri implantation milieu influencing fertilization and embryo development. PMID- 11912282 TI - Regulation of natural antibiotic expression by inflammatory mediators and mimics of infection in human endometrial epithelial cells. AB - The natural antibiotic molecules, beta-defensins 1 and 2 (HBD1/2) and secretory leukocyte protease inhibitor (SLPI), have an important role in mucosal defence and are present in the uterus. This study details their regulation in primary endometrial epithelial cells and in two endometrial cell lines (MFE/HES). Cells were treated with proinflammatory molecules and mimics of infection [lipopolysaccharide (LPS) and lipoteichoic acid (LTA)]. mRNA for HBD1, HBD2 and SLPI was detected in primary endometrial epithelial cells using real-time quantitative PCR. HBD1 mRNA was present at very low levels preventing conclusive study of its regulation. However, HBD2 mRNA expression was increased by interferon-gamma, interleukin (IL)-1beta alone and IL-1beta+tumour necrosis factor (TNF)-alpha. SLPI mRNA was not affected by proinflammatory mediators, although protein levels fell in the presence of IL-1beta+TNFalpha. LPS had little effect on antimicrobial expression. However, there was a trend towards increased expression with LTA treatment for 4-8 h. Antimicrobial expression in endometrial cell lines was similar to that in primary cells, although SLPI was increased by IL-1beta+TNFalpha treatment. These results suggest that in endometrium some natural antibiotics (e.g. SLPI) may be constitutively expressed providing a basal level of protection, while others (e.g. HBD2) are inducible allowing maximal antimicrobial activity during infection. Natural antimicrobials will have an important role in endometrium in protecting against infection. PMID- 11912283 TI - Pleiotrophin (PTN) and midkine (MK) mRNA expression in eutopic and ectopic endometrium in advanced stage endometriosis. AB - Endometriosis is characterized by the ectopic implantation of endometrium on peritoneal surfaces. Angiogenic and growth factors may play a significant role in the pathogenesis of endometriosis. Midkine (MK) and pleiotrophin (PTN) are two related peptides associated with carcinogenesis and angiogenesis. To test the hypothesis that a higher expression of MK and PTN in ectopic and eutopic endometrium from women with endometriosis might favour increased angiogenesis and growth with subsequent ectopic implantation, we investigated PTN and MK expression by quantitative competitive PCR (QC-PCR) in endometrium from 30 women with severe, stages III and IV endometriosis and from 30 women without endometriosis. Total RNA was extracted and reverse transcribed into cDNA, and QC PCR was performed to evaluate PTN and MK mRNA expression. Results were analysed by analysis of variance. Eutopic endometrium from endometriosis patients showed increased expression of MK and PTN mRNA compared with endometrium from normal women in the luteal phase (P < 0.05). MK and PTN mRNA expression in ectopic endometrium was significantly lower than that in eutopic endometrium from women with and without endometriosis (P < 0.05). Our results suggest increased MK and PTN expression may be related to the initiation of ectopic endometrial implants and peritoneal invasion. PMID- 11912284 TI - Expression of prolactin-releasing peptide and its receptor in the human decidua. AB - Human decidua and decidualized endometrial cells produce prolactin (PRL). Several growth factors and cytokines have been shown to regulate decidual PRL release, but a specific PRL-releasing substance remains to be characterized. Prolactin releasing peptide (PrRP) is a peptide isolated from the brain and distinguished by its potent and specific stimulation of PRL release by cultured pituitary cells. Here, we demonstrate that human decidua expresses immunoreactive PrRP as well as the mRNAs encoding PrRP and its receptor. First trimester deciduas were obtained from women undergoing elective termination of pregnancy. Tissue specimens were stained by immunohistochemistry using a rabbit anti-human PrRP-31 antibody, and PrRP was localized in both epithelial cells of the decidual glands and in stromal cells, with diffuse distribution and no special relation with the neighbourhood of blood vessels. In primary cultures of decidual stromal cells, PrRP and PrRP receptor gene expression were detected using RT-PCR, and the identity of the PCR products was further confirmed by restriction enzyme digestion. The effect of PrRP on decidual PRL release was also evaluated, and there was a significant increase in PRL production (135 +/- 4% of control levels, P < 0.05) after incubation of decidual stromal cells with synthetic PrRP. The expression of PrRP and PrRP receptor in human decidual cells and the ability of PrRP to induce PRL secretion by cultured decidual cells suggests that this peptide may be a novel local modulator of decidual PRL release. PMID- 11912285 TI - Expression of activin receptors, follistatin and betaglycan by human endometrial stromal cells; consistent with a role for activins during decidualization. AB - Decidualization of the human endometrium is critical for implantation, but the mechanisms involved are largely unknown. Activin subunits are expressed in endometrium during decidualization. From its known actions in cell differentiation and tissue remodelling, we hypothesized that activin A is involved in the paracrine regulation of decidualization. We examined the expression of activin receptors (ActRs) by semi-quantitative and real-time RT PCR. mRNA for all ActR subtypes (Ia, Ib, IIa and IIb) was detected in endometrium, with maximal expression in the early secretory phase and in early pregnancy. ActR protein was localized exclusively to stromal and endothelial cells. This expression pattern was confirmed by in-situ hybridization. Activin bioavailability is locally regulated by its binding protein, follistatin, and also by the antagonist, inhibin. Inhibin competition for ActRII binding is enhanced by the binding protein, betaglycan. Follistatin and betaglycan were also detected in the endometrium, localized to stromal and epithelial cells. This co expression of activin subunits, receptors and binding proteins indicates that stromal cells are capable of responding to activin, and that there is tight local regulation of activin action within the endometrium. As activin production is up regulated in decidual cells, this provides further evidence for an involvement of activins during stromal cell decidualization. PMID- 11912286 TI - Hormonal and embryonic regulation of chemokines IL-8, MCP-1 and RANTES in the human endometrium during the window of implantation. AB - Chemokines are a family of small polypeptides which specialize in the attraction of leukocytes. The presence of specific leukocyte subsets at the implantation site is an important element of the complex, and not completely understood, process of embryonic implantation. This report includes the investigation of the in-vivo immunolocalization and hormonal regulation of interleukin (IL)-8, monocyte chemotactic protein (MCP)-1 and RANTES (regulated upon activation normal T-cell expressed and secreted) in the human endometrium during hormone replacement therapy cycles for oocyte recipients in an IVF programme. In addition, we have analysed the embryonic regulation of these endometrial epithelial chemokines (IL-8 and MCP-1) using an in-vitro model for the apposition phase of human implantation by co-culturing single human embryos until the blastocyst stage with human endometrial epithelial cells (EEC). IL-8 and MCP-1 were immunolocalized in the human endometrium to the glandular and lumenal epithelium as well as to the endothelial cells. RANTES was mainly localized to the stromal compartment and endothelial cells. The immunoreactive levels of endometrial IL-8 and MCP-1 were up-regulated by the administration of progesterone during the receptive phase of the cycle. Furthermore, it was demonstrated that, in vitro, the human blastocyst does not produce measurable amounts of IL-8, MCP-1 or RANTES; however, it does up-regulate EEC IL-8 mRNA expression (P < 0.05) and protein production (P < 0.05), but not IL-8 secretion. The human embryo did not regulate EEC MCP-1 expression. These results provide evidence of hormonal and embryonic regulation of specific endometrial chemokines, suggesting two different but related mechanisms to induce the production of chemokines by the EEC, thus contributing to the attraction of specific leukocyte populations during the peri-implantation phase. PMID- 11912287 TI - Localization of indoleamine 2,3-dioxygenase in human female reproductive organs and the placenta. AB - Indoleamine 2,3-dioxygenase (IDO) has been implicated in regulation of feto maternal tolerance and protection against intracellular and extracellular pathogens. We have studied the expression of IDO in the human female reproductive tract and the placenta by immunohistochemistry. Endometrial glandular and surface epithelial cells showed increasing IDO expression during the course of the menstrual cycle. In term placenta, IDO was irregularly localized to the mesenchymal core and found in isolated areas of the syncytiotrophoblast. In first trimester pregnancy, IDO was not present in placental villi, but was present in glandular epithelium of the decidua, and there were distinctly positive cells scattered in the connective tissue, sometimes in conjunction with lymphoid aggregates. The endothelium of spiral arteries and of capillaries showed some, albeit no generalized, reactivity. IDO was also present in the epithelium of cervical glands and of Fallopian tubes. Specificity of antibody binding was confirmed by Western blot analysis. IDO mRNA was detected in first trimester decidua as determined by RT-PCR. IDO is secreted, as determined by analysis of cervical mucus by high pressure liquid chromatography for the presence of the tryptophan metabolite L-kynurenine, indicating IDO activity. Our results support the concept of IDO providing a mechanism of innate immunity protecting against ascending infections in the female reproductive tract. PMID- 11912288 TI - Reduced expression of tissue inhibitor of metalloproteinase (TIMP)-2 in gestational trophoblastic diseases. AB - To elucidate the involvement of type IV collagenases [matrix metalloproteinase (MMP)-2 and MMP-9] and their tissue inhibitors (TIMP-1 and TIMP-2) in the development of gestational trophoblastic disease (GTD), we quantified their levels in hydatidiform mole and choriocarcinoma tissues using specific enzyme linked immunosorbent assays, and the results were compared with those from normal first trimester placenta. Levels of pro-MMP-2 were increased in hydatidiform mole, and they were further elevated in choriocarcinoma. Levels of pro-MMP-9 in choriocarcinoma and those of TIMP-1 in both hydatidiform mole and choriocarcinoma were also increased. In contrast, TIMP-2 levels were markedly decreased in both hydatidiform mole and choriocarcinoma. Similar results were obtained by the tissue culture of first trimester placenta and hydatidiform mole. Gelatin zymography indicated that the levels of both pro- and activated forms of MMP-2 and MMP-9 were higher in hydatidiform mole and choriocarcinoma. The decreased expression of TIMP-2 in hydatidiform mole and choriocarcinoma was confirmed by Western blot, Northern blot and immunohistochemistry, with the decrease being more pronounced in choriocarcinoma. Taken together, the present study shows that both TIMP-2 mRNA and protein levels are markedly decreased in GTD and the imbalance of MMP-TIMP production, shifted toward greater MMP activity, may be involved in the pathogenesis of GTD. PMID- 11912291 TI - Introduction: Molecular topics in dialysis. PMID- 11912289 TI - Use of cDNA arrays to generate differential expression profiles for inflammatory genes in human gestational membranes delivered at term and preterm. AB - Inflammatory processes are implicated in preterm labour (PTL). To identify potential novel markers for PTL, we have used commercial cDNA arrays to generate profiles of differential expression of inflammation-associated genes in gestational membranes with term and PTL. RNA for cDNA probe synthesis was isolated from reflected human amnion and choriodecidua membranes delivered following Caesarean section at term before the onset of labour (TNL, n = 4), spontaneous labour at term (TSL, n = 4), and PTL with and without chorioamnionitis (PTL(+INF) and PTL(-INF) respectively, n = 4 each). Profiles were displayed relative to TNL and statistical comparisons of TSL versus TNL and PTL(+INF) versus PTL(-INF) were performed. Elevated expression of chemokines macrophage inflammatory protein 1beta(MIP-1beta) and pulmonary and activation regulated chemokine (PARC) was observed in PTL(+INF) compared to PTL(-INF) amnion and choriodecidua respectively (P = 0.03). Likewise, the cytokines oncostatin-M and pre-B cell enhancing factor (PBEF) were more highly expressed in PTL(+INF) compared with PTL(-INF) and in TSL compared with TNL respectively (P = 0.03). Conversely, inhibin A, tissue inhibitors of matrix metalloproteinase (TIMP)-3 and TIMP-4 were all significantly elevated in PTL(-INF) compared with PTL(+INF) (P = 0.03). Furthermore, differential expression patterns of classes of genes, grouped according to function (e.g. chemokines), were noted. The cDNA array approach holds promise for identification of new candidate markers or combinations thereof for prediction or diagnosis of PTL, as well as for increasing our understanding of the particular aetiologies involved. PMID- 11912292 TI - Molecular genetics in renal medicine: what can we hope to achieve? PMID- 11912293 TI - Morphological and functional changes in the dialysed peritoneal cavity: impact of more biocompatible solutions. AB - Loss of peritoneal function is a major factor leading to treatment failure in peritoneal dialysis (PD). To date, however, the relationship between the observed functional changes (reduction in ultrafiltration and changes in solute transport) and the structural alterations in the membrane have not been fully defined. Here we present data from the Peritoneal Biopsy Registry identifying and characterizing both changes in parietal peritoneal membrane thickness (degree of fibrosis) and vascular alterations (blood vessel degenerative changes) and relate these to the duration of dialysis. The genesis of functional changes in the membrane may be related to these vascular alterations. This issue is discussed in relation to the importance of nitric oxide and its synthetic enzymes in this process and its potential interaction with endothelial cell aquaporin function. It is widely believed that conventional acidic, lactate-buffered glucose containing dialysis solutions contribute to both the structural and functional changes in the dialysed peritoneal membrane. The introduction of new more biocompatible solutions potentially allows us to reverse or attenuate these negative changes. This will be discussed in the context of our current understanding of peritoneal pathology in PD. PMID- 11912295 TI - The acute phase response in chronic haemodialysis patients: a marker of cardiovascular disease? PMID- 11912294 TI - Clinical advantages of new peritoneal dialysis solutions. AB - A review is given of the various mechanisms by which conventional glucose/lactate based peritoneal dialysis solutions can induce damage to the peritoneal membrane. The potential advantages of newly developed dialysis solutions and the results of recent studies on their use in patients are discussed. PMID- 11912296 TI - Haemodialysis, atherosclerosis and inflammation-identifying molecular mechanisms of chronic vascular disease in ESDR patients. PMID- 11912297 TI - Determinants of uraemic toxin removal. AB - The inability to excrete a wide spectrum of toxins is one of the hallmarks of uraemia and is responsible for many of the signs and symptoms of this syndrome. Therefore, an understanding of the factors influencing uraemic toxin removal, during both the pre-dialysis and dialytic phases, is critically important. This review addresses many of these factors, which include the importance of preserved residual renal function and the effect of therapy frequency. In addition, specifically with respect to larger uraemic toxins, the importance of convection and treatment duration is discussed. All of these issues will be major considerations as new dialytic therapies for end-stage renal disease patients are developed and implemented. PMID- 11912330 TI - A comparison of problem-based and conventional curricula in nursing education. AB - The purpose of this study was to compare graduating baccalaureate students in a problem-based curriculum with those in a conventional nursing program with regard to perceived preparation for clinical practice, clinical functioning, knowledge and satisfaction with their education. Prior to graduation, students completed a self-report questionnaire that consisted of five sections and took about 45 minutes to complete. Following graduation, their pass rates on the National Nursing Registration Examination (RN Exam) were also compared. The findings indicated no significant differences in their perceived preparation for nursing practice, although the conventional students scored higher in all areas. There were also no significant differences between the two groups in their perceived clinical functioning, although there was a trend toward higher function in the areas of communication and self-directed learning in the PBL group. There were no statistically significant differences in RN scores. The PBL students scored significantly higher on perceptions of their nursing knowledge, particularly in the areas of individual, family and community health assessment, communication, teaching/learning, and the health care system. The students undertaking the PBL program were more satisfied with their educational experience than their counterparts in the conventional program, indicating higher satisfaction with tutors, level of independence, assessment and program outcomes, but no difference in relation to workload or clarity of expectations. This study contributes to our understanding of the relationship between different educational approaches and student outcomes. It suggests that PBL is an effective approach for educating nurses. Furthermore, it indicates that nursing students in the PBL program, like their counterparts in PBL medical programs, report higher levels of satisfaction. Future studies that are longitudinal in design and rely less on self-report measures would contribute further to our understanding of the benefits and limitations of PBL in nursing education. PMID- 11912331 TI - Accuracy of student self-assessment ability compared to their own performance in a problem-based learning medical program: a correlation study. AB - OBJECTIVE: The purpose of this study was to evaluate the accuracy of self assessment ability of students enrolled in a Problem-based Learning program. METHODS: Seventy students enrolled in their third year of a four-year program were invited to voluntarily participate in the study. Self-assessment questionnaire was used to measure the students' self-assessment ability on two different occasions: 1) prior to standardised oral examination in predicting their performance, and 2) following the examination estimating their performance. The accuracy of the self-assessment was investigated by the relation between self assessment and performance of the students. RESULTS: Our study showed that self assessment pre-examination was not accurate compared to performance at the oral examination (r ranging from 0.042 to 0.243). However, accuracy is slightly better when the student self-assesses his performance a posteriori, but the relation stays very low (r ranging from 0.257 to 0.334). CONCLUSION: According to our results, the students in the third year of a self-directed Problem-based Learning medical four year program demonstrated poor accuracy of the self-assessment when compared to their own performance. PMID- 11912332 TI - Longitudinal and concentrated communication skills programmes: two dutch medical schools compared. AB - AIM OF THE STUDY: The communication skills of students of the Dutch medical schools of Maastricht and Leiden were compared, to assess the effectiveness of these schools' different approaches to communication skills training. Both schools have a six-year undergraduate medical curriculum, divided into four preclinical years and two years of clinical clerkships. The Maastricht problem based curriculum offers an integrated clinical skills training programme, including communication skills, which runs throughout the first four years. Communication skills training in Leiden is concentrated in courses in the preclinical phase, at the beginning of the clinical phase and preceding two clerkships. METHOD: Communication skills of fourth-year and sixth-year students (N = 161) of both universities were assessed using four OSCE stations in which students did entire consultations with standardised patients. Trained observers rated these consultations, using a checklist. RESULTS: Maastricht students of both year groups obtained significantly higher checklist scores for their communication skills than their Leiden colleagues. The Leiden students' scores increase between years 4 and 6, whereas those of the Maastricht students showed no significant change. DISCUSSION: The higher scores obtained by the Maastricht students indicate a greater overall effectiveness of a longitudinal, integrated approach compared with concentrated courses. Absence of formal training in the clinical phase in Maastricht leads to stabilisation of communication skills, whereas the increase in the Leiden students' scores between years 4 and 6 offers evidence that formal communication skills training during the clinical phase does pay off. These findings suggest that the preferred approach to communication skills training would be an integrated, longitudinal programme, which continues during the clinical years. PMID- 11912333 TI - Correlation of problem-based learning facilitators' scores with student performance on written exams. AB - Problem-based learning (PBL) is widely used in medical education. In some cases, facilitators assign a grade to reflect a student's performance in small-group sessions. In our PBL track, facilitators were asked to assess student knowledge base independent of their group participatory skills. To determine if facilitators' grades were correlated with student performance in written exams, a retrospective study of data from our PBL track was undertaken. Data from 156 students and 107 facilitators in six years of a PBL track at Penn State College of Medicine was analyzed by the Pearson correlation after pairing facilitator grades with written exam grades for each of the eight blocks of the curriculum. Exam reliability and validity were assessed by Cronbach's alpha and correlation with USMLE I board scores. The mean alpha was 0.549 +/- 0.221. The mean correlation with USMLE scores was 0.558 +/- 0.151. Facilitators' scores for knowledge were positively associated with students' exam grades. The corresponding significant Pearson correlation coefficients were between 0.342 0.622. However, the coefficients of determination showed that the correlation was not significant. Coefficients of determination showed that the knowledge scores explained only 12 to 39% of the variance in exam scores. Overestimation by facilitators was significantly (p < 0.0001) greater for students in the bottom 25% of the class by exam score than for students in the top 25% of the class. On the basis of this study, we concluded that facilitator assessment of student knowledge base is not useful. PMID- 11912335 TI - What does two disciplines of scientific psychology have to say to medical education? PMID- 11912336 TI - Measuring self-assessment: current state of the art. AB - The competent physician pursues lifelong learning through the recognition of deficiencies and the formulation of appropriate learning goals. Despite the accepted theoretical value of self-assessment, studies have consistently shown that the accuracy of self-assessment is poor. This paper examines the methodological issues that plague the measurement of self-assessment ability and presents several strategies that address these methodological problems within the current paradigm. In addition, the article proposes an alternative conceptualization of self-assessment and describes its associated methods. The conclusions of prior research in this domain must be re-examined in light of the common pitfalls encountered in the design of the studies and the analyses of the data. Future efforts to elucidate self-assessment phenomena need to consider the implications of this review. PMID- 11912337 TI - Review of Papillostrongylus Johnston & Mawson, 1939 (Nematoda: Strongyloidea) from wallabies and kangaroos (Marsupialia: Macropodidae) using morphological and molecular techniques, with the description of P. barbatus n. sp. AB - The strongyloid nematode genus Papillostrongylus Johnston & Mawson, 1939, from kangaroos and wallabies, is reviewed using morphological and molecular methods. P. labiatus Johnston & Mawson, 1939 is re-described from material from the type host, the black-striped wallaby Macropus dorsalis, from eastern Queensland, Australia, in which it is a relatively common parasite. Additional records from M. parryi and Thylogale thetis are confirmed and considered to represent examples of host-switching. A geographically disjunct population of the nematode species occurs in M. bernardus and Petrogale brachyotis in Arnhem Land, Northern Territory, but assessment of its status requires additional material. Nematodes from M. rufus, M. giganteus, M. fuliginosus and M. robustus from inland regions of Australia, formerly attributed to P. labiatus, are here assigned to a new species, P. barbatus, distinguished by the presence of an external leaf-crown, larger size, by greater spicule length in the male and by a sinuous vagina in the female. Additional hosts of P. barbatus n. sp. are Petrogale assimilis and Pet. lateralis purpureicollis. Sequence analyses of the second internal transcribed spacer of ribosomal DNA (ITS-2) also showed that P. barbatus n. sp. differed at 40 (16.7%) of the 240 alignment positions when compared with P. labiatus. Most of these interspecific sequence differences occurred in loops or bulges of the predicted precursor rRNA secondary structure, or represented partial or total compenstory base pair changes in stems. PMID- 11912338 TI - Taxonomy of trichodinids from the gills of marine fishes in coastal regions of the Yellow Sea, with descriptions of two new species of Trichodina Ehrenberg, 1830 (Protozoa: Ciliophora: Peritrichia). AB - Seven species of marine fishes from coastal regions of the Yellow Sea were examined for ectoparasitic trichodinids (Protozoa, Ciliophora). A total of seven species belonging to the genus Trichodina Ehrenberg, 1830, including two new species, occurred as gill parasites. These were T. rectispina n. sp., T. circinantis n. sp., T. frequentis Shtein, 1979, T. jarmilae Lom & Laird, 1969, T. hexagrammi Zhukov, 1964, T. minima Shtein, 1979 and T. ovonucleata Raabe, 1958. Each was investigated following silver nitrate and protargol impregnation. Morphometric data and comparative descriptions of these trichodinids are provided along with details of their prevalence and intensity of infestation. PMID- 11912340 TI - Steganoderma (Steganoderma) valchetensis n. sp. (Digenea: Zoogonidae) from the relict fish Gymnocharacinus bergi (Osteichthyes: Characidae) in Argentina. AB - A new species of Steganoderma (Steganoderma) (Digenea: Zoogonidae) was found in the digestive tract of the southernmost, endemic and scale-less characid, Gymnocharacinus bergi, inhabiting thermal headwaters of the Valcheta Creek in northern Patagonia, Argentina. Steganoderma valchetensis n. sp. is included in the subgenus Steganoderma because of having a sessile ventral sucker and caeca reaching to the testes. This new species can be distinguished from other species of this genus on the basis of body-length, sucker-ratio, the length of the cirrus sac, the morphology of the seminal vesicle and gonads, and the location of the vitelline follicles and genital pore. The taxonomic status of the species of Steganoderma, parasitising the freshwater fishes of Patagonia, in southern Argentina, and Chile, is also discussed. PMID- 11912339 TI - Redescription of Zoniolaimus mawsonae Beveridge, 1983 (Nematoda: Strongyloidea) and the description of Z. latebrosus n. sp. from the red kangaroo Macropus rufus (Marsupialia: Macropodidae) based on morphological and molecular data. AB - This study shows that the description of Z. mawsonae given by Beveridge (1983) represented a composite of two species. Hence, Z. mawsonae Beveridge, 1983 is re described and a new species, Z. latebrosus, is described. Males of the two species differ in the lengths of their spicules (0.94-1.23 mm in Z. mawsonae compared with 1.53-1.95 mm in Z. latebrosus) and in several characteristics of the bursa and genital cone. Females of the two species can be identified based on the shape of the posterior end of the body, that of Z. mawsonae being markedly swollen. Molecular data, obtained from single strand conformation polymorphism (SSCP) analysis and DNA sequencing of the second internal transcribed spacer of ribosomal DNA, provided additional support for the separation of the two species. PMID- 11912341 TI - An SEM study of Phocascaris cystophorae Berland, 1964 (Nematoda: Anisakidae), a parasite of the hooded seal Cystophora cristata. AB - Scanning electron microscopy was used to study the surface morphological characteristics (excretory pore, interlabia knobs, lips and adjacent structures, caudal papillae and papilla-like structures) of the nematode Phocascaris cystophorae, a parasite from the stomach of the hooded seal Cystophora cristata. A comparative morphological analysis was made between species of Phocascaris and Contracaecum osculatum (sensu lato), which are all parasitic in phocid seals. PMID- 11912343 TI - Description of Tylenchorhynchus shimizui n. sp. from Paraguay and notes on T. leviterminalis Siddiqi, Mukherjee & Dasgupta from Japan (Nematoda: Tylenchida: Telotylenchidae). AB - A new species of Tylenchorhynchus Cobb, 1913 from Paraguay, T. shimizui n. sp., and T. leviterminalis Siddiqi, Mukherjee & Dasgupta, 1982 from Japan are described and illustrated. T. shimizui n. sp. is a monosexual species characterised by females with a medium-sized body, L=0.70-0.82 mm, stylet 18.4 20.8 microm long, a rounded lip region slightly set off from the body contours bearing six annules, oesophageal glands slightly overlapping the intestine laterally and a subcylindrical tail with a bluntly pointed smooth terminus. A population of T. leviterminalis associated with sugarcane from Japan is also described and compared with previous descriptions. This is the first record for T. leviterminalis in Japan. Spores of a Pasteuria species were observed filling the body-cavity of some specimens of T. leviterminalis. PMID- 11912342 TI - Genetic markers in the study of Anisakis typica (Diesing, 1860): larval identification and genetic relationships with other species of Anisakis Dujardin, 1845 (Nematoda: Anisakidae). AB - Genetic variation at 21 gene-enzyme systems was studied in a sample of an adult population of Anisakis typica (Diesing, 1860) recovered in the dolphin Sotalia fluviatilis from the Atlantic coast of Brazil. The characteristic alleles, detected in this population, made it possible to identify as A. typica, Anisakis larvae with a Type I morphology (sensu Berland, 1961) from various fishes: Thunnus thynnus and Auxis thazard from Brazil waters, Trachurus picturatus and Scomber japonicus from Madeiran waters, Scomberomorus commerson, Euthynnus affinis, Sarda orientalis and Coryphaena hippurus from the Somali coast of the Indian Ocean, and Merluccius merluccius from the Eastern Mediterranean. Characteristic allozymes are given for the identification, at any life-stage and in both sexes, of A. typica and the other Anisakis species so far studied genetically. The distribution of A. typica in warmer temperate and tropical waters is confirmed; the definitive hosts so far identified for this species belong to delphinids, phocoenids and pontoporids. The present findings represent the first established records of intermediate/paratenic hosts of A. typica and extend its range to Somali waters of the Indian Ocean and to the Eastern Mediterranean Sea. A remarkable genetic homogeneity was observed in larval and adult samples of A. typica despite their different geographical origin; interpopulation genetic distances were low, ranging from D(Nei)=0.004 (Eastern Mediterranean versus Somali) to D(Nei)=0.010 (Brazilian versus Somali). Accordingly, indirect estimates of gene flow gave a rather high average value of Nm = 6.00. Genetic divergence of A. typica was, on average, D(Nei)=1.12 from the members of the A. simplex complex (A. simplex s.s, A. pegreffii, A. simplex C) and D(Nei)=1.41 from A. ziphidarum, which all share Type I larvae; higher values were found from both A. physeteris (D(Nei)=2.77) PMID- 11912344 TI - A cladistic analysis of the tribe Labiostrongylinea Beveridge, 1983 (Nematoda: Cloacinidae) parasitic in macropodoid marsupials (Marsupialia: Macropodoidea), with a redescription of Parazoniolaimus collaris Johnston & Mawson, 1939. AB - Morphological characters of both adult and juvenile representatives of the genera comprising the Labiostrongylinea (Strongylida: Cloacinidae: Cloacininae) were examined to prepare an amended description of Paralabiostrongylus bicollaris, a redescription of Parazoniolaimus collaris and a cladistic analysis of the tribe. A cluster of Wallabinema spp., with a fleshy collar on which amphids and deirids are borne, was used as the outgroup for the analysis. The 50% consensus tree, bootstrapped through 1000 replicates, derived from the cladistic analysis had a basal polytomy of Dorcopsinema and Paralabiostrongylus; and a crown polytomy of Labiosimplex, Labiomultiplex, Parazoniolaimus, Labiostrongylus and Potorostrongylus. This analysis provided reasonable evidence for considering all 7 clades as being of generic status and amended diagnoses were prepared. The morphology of the cephalic end of fourth stage juveniles, cephalic papillae and amphids lying posterior to the lips, gave support to the contentions that a fleshy collar is a plesiomorphic character and that Dorcopsinema is the most primitive genus within the group. There was, however, insufficient support from morphological data to provide a robust resolution of generic relationships. Host and biogeographical data were indicative of Paralabiostrongylus and Labiosimplex having been derived from ancestral Dorcopsinema, then Labiostrongylus, Labiomultiplex and Parazoniolaimus derived from ancestral Labiosimplex, and Potorostrongylus from ancestral Labiomultiplex. Additional data, derived from molecular sequencing, is needed to help resolve the relationships within the tribe. PMID- 11912345 TI - Variability in the taxonomic characters of Black Sea gyrodactylids (Monogenea). AB - Variability of the haptoral hard parts of four Black Sea Gyrodactylus species (G. alviga Dmitrieva & Gerasev, 2000; G. crenilabri Zaika, 1966; G. flesi Malmberg, 1957; G. sphinx Dmitrieva & Gerasev, 2000) was studied in relation to season, host and geographical variability. A discriminant analysis was carried out on the measurements of 14 characters of the haptor of all four species. The haptoral hard parts of the investigated gyrodactylids decrease in size with an increase in water temperature. It is known that life-span of gyrodactylids is negatively correlated with water temperature and they exhibit peak intensities at low temperature. At the same time, life-span is considered to be a general indicator of a favourable environment. Therefore, high water temperature is not apparently advantageous for gyrodactylids. In the case of low marine salinity, in adapted species the sizes of the haptoral hard parts are larger at lower levels of salinity and vice versa for typical marine species. Lastly, in specimens parasitising the primary host, these features are larger than in those parasitising a secondary host. It is possible to say that, in general, the more favourable the environment for gyrodactylids the larger their haptoral hard parts. Unfavourable environmental conditions reduce the time of embryogenesis, which has a dual effect on gyrodactylids: positive in relation to their reproduction (by increasing the reproduction rate) and negative in relation to their development (by decreasing the size of the haptoral hard parts). PMID- 11912346 TI - Plagiorhynchidae Meyer, 1931 (Acanthocephala) from Australasian birds and mammals, with descriptions of Plagiorhynchus (Plagiorhynchus) menurae (Johnston, 1912) and P. (P.) allisonae n. sp. AB - New host and locality records are given for Plagiorhynchus (Plagiorhynchus) charadrii (Yamaguti, 1939) Van Cleave, 1951 and P. (Prosthorhynchus) cylindraceus (Goeze, 1782) Schmidt & Kuntz, 1966. The uncertainty of identification of a plover host of P. (P.) charadrii as well as the origins of P. (P.) cylindraceus (found in Australia but not New Zealand) and its occurrence in both bird and mammal hosts are discussed. P. (P.) menurae (Johnston, 1912) Golvan, 1956 is redescribed, including the male, and new host, Menura alberti Bonaparte, and locality records are given. P. (P.) allisonae n. sp. is described from Haematopus ostralegus finschi (Martens) and H. unicolor (Forster) from the South Island of New Zealand. P. (P.) allisonae can be differentiated from its congeners by having a proboscis armature of 18-23 rows of 14-20 hooks, thorns of hooks shorter than simple roots with short manubria, eight tubular cement glands and eggs of 134-154 x 33-36 microm in size. The presence of P. (P.) gracilis Petrochenko, 1958 in Australia is questioned. New host and locality records are given for Porrorchis hylae (Johnston, 1914) Schmidt & Kuntz, 1967 and the northern distribution of P. hydromuris (Edmonds, 1957) Schmidt & Kuntz, 1967 confirmed. PMID- 11912347 TI - Trimusculotrema schwartzi n. sp. (Monogenea: Capsalidae) from the skin of the stingray Dasyatis zugei (Elasmobranchii: Dasyatidae) off Hong Kong, China. AB - Trimusculotrema schwartzi n. sp. (Capsalidae) is described from the skin of the stingray Dasyatis zugei (Elasmobranchii: Rajiformes: Dasyatidae) off Hong Kong, China. Only three other species have been placed in the genus Trimusculotrema: T. micracantha (Euzet & Maillard, 1967), T. leucanthemum (Euzet & Maillard, 1967), and T. uarnaki Whittington & Barton, 1990. T. schwartzi n. sp. may be differentiated from all known species of Trimusculotrema by the length of the anterior hamuli and by the absence of pigment shields over the eye-spots. Its occurrence on a stingray off China represents a northern extension of the geographical range of Trimusculotrema. PMID- 11912348 TI - Three species of plagioporine opecoelids (Digenea), including a new genus and two new species, from marine fishes from off the coast of Chile. AB - Villarrealina peruanus (syn. Pirupalkia queulensis) is described from Cilus gilberti, off the Talcahuano region, Chile. It is considered an opecoelid and is distinguishable by the combination of its extensive uterus, which reaches to the posterior extremity, and the vitellarium restricted to the hindbody. Jerguillicola leonora n. g., n. sp. from Aplodactylus punctatus, off the Talcahuano region, Chile, is distinguished by the uterus reaching into the post testicular region, the lack of a seminal receptacle, the eggs attached together is strings formed by a matrix, the large cirrus-sac filled mainly with fairly dense connective tissue, a long coiled seminal vesicle and a long coiled ejaculatory duct. Neolebouria georgenascimentoi n. sp. from Pinguipes chilensis and Prolatilus jugularis, off the Talcahuano region, Chile, can be distinguished from its congeners by a its elongate cirrus-sac and oblique entire to weakly lobed testes. PMID- 11912349 TI - Cadaveric and clinical study of endoscope-assisted microneurosurgery for cerebral aneurysms using angle-type rigid endoscope. AB - Highly advanced optical equipment enables endoscopic surgery to be performed in neurosurgery. We developed an angle-type rigid endoscope having an angled shaft, and has performed endoscope-assisted surgery in neck clipping of 50 cerebral aneurysms. Anatomical study concerning the perforating branch was made through a pterional approach using 3 cadavers. By using endoscope, we can observe the posterior communicating artery and anterior choroidal artery behind the internal carotid artery without retract the internal carotid artery and the surrounding structures. Furthermore IIIrd~VIIIth cranial nerve in opposite side and basilar artery in the posterior fossa can be observed. As the clinical study, we used endoscope in 50 clipping of cerebral aneurysms (29 Internal carotid artery aneurysms, 6 anterior communicating artery aneurysms, 2 anterior cerebral artery aneurysms, 7 middle cerebral artery aneurysms, 5 vertebral artery-posterior inferior cerebellar artery aneurysm, and basilar bifurcation aneurysms). The endoscopic image showed in incomplete clipping in 2 cases, where additional clipping was then performed. In all subject, complete clipping was performed using both microscopic and endoscopic view. Use of the angle-type rigid endoscope together with Doppler ultrasound, should increase the feasibility of complete clipping procedure, resulting in reduced complication rate. PMID- 11912350 TI - Spiral artery of placenta: development and pathology-immunohistochemical, microscopical, and electron-microscopic study. AB - The spiral artery (=SA) is an important muscular artery, which controls the blood volume to the placenta. Preeclampsia is thought to be induced by the failure of the placenta by dysfunction of SA. To clarify the function of SA, we examined forty-eight placentae and its morphological and biological characteristics: 36 normal placentae and 12 placentae with preeclampsia. Gestational age of normal placentas was between 19 and 40 weeks and placentae with preeclampsia was between 31-36 weeks. The wall of the placental segment of SA by both light and electron microscope, and the wall width of SA and gestation age were compared each other. The wall of SA, with the invasion of trophoblast, was thin, but SA without trophoblasts was thick in width. At normal placenta, the diameter of SA was dilative constantly, but the width of the wall showed a tendency of getting thinning as advances. Ultrastructually, we found the trophoblast of thin wall of SA with dilated lumen. These ultrastructual alternations were consistence with the light microscopical findings. In preeclampsia, the lumen of SA between normal pregnancy and one with preeclampsia was almost same, but the wall width was thick, compared with normal pregnancy (P<0.05). We concluded that trophoblastic invasion control the functions of SA. PMID- 11912351 TI - Molecular genetics of spinal muscular atrophy: contribution of the NAIP gene to clinical severity. AB - Spinal muscular atrophy (SMA) is one of the most common autosomal recessive disorders characterized by degeneration of anterior horn cells in the spinal cord, and leads to progressive muscular weakness and atrophy. At least three SMA related genes have been identified: SMN1, NAIP and p44t. We analyzed these genes in 32 SMA patients and found that the SMN1 gene was deleted in 30 of 32 patients (94 %), irrespective of clinical type. The NAIP gene was deleted in 6 patients and its deletion rate was higher in type I patients than that in type U or V. Further, in type I patients lacking the NAIP gene, deterioration in their respiratory function is more rapid than in those type I patients retaining the NAIP gene. Since complete p44t deletion was observed in only 3 patients, the correlation between the p44t deletion and severity of SMA remained ambiguous. We concluded that the NAIP deletion was closely related to the clinical severity of SMA and was a predictive marker of SMA prognosis, while the SMN1 deletion did not correlate with clinical severity. PMID- 11912352 TI - Outcome analysis of reoperations after lumbar discectomies: a report of 22 patients. AB - Our aim was to analyse causes for persistence of pain after lumbar discectomy and outcome of reoperations. Out of 37 reoperated patients, 22 with a minimum follow up period of one year were included in this retrospective study concerning the years 1993 to 2000. All patients had previously undergone laminotomy and discectomy and fusion was not required during second operations. Outcome was evaluated according to the modified criteria of Kawabata et al. Overall incidence of reoperation was 6.5%. Reoperations were performed because of recurrent disc herniation in 9 patients, epidural fibrosis in 8, and de novo disc herniation at a different level in 5. Contrast enhanced computerized tomography was used in 17 patients and this might be the reason for misdiagnosis of recurrent disc herniation in the two patients with epidural fibrosis. In patients with de novo disc herniation, symptoms recurred earlier. In 20 patients, satisfactory relief of pain, as well as better outcome could be achieved (p<0.05), but no significant improvement in neurological deficits was observed. Excellent results were obtained more in patients with recurrent disc herniation and poor outcomes correlated with long (> 1 year) time intervals for onset of recurrent sciatica (p<0.05). However, patients with epidural fibrosis were also glad postoperatively for decreased pain severity. Only co-existence of epidural fibrosis and de novo disc herniation predicted an inferior outcome. Although recurrent disc herniation seemed to respond best to surgical treatment, we recommend reoperation when objective preoperative findings indicate the presence of surgically correctable compression regardless of its type. PMID- 11912354 TI - In vivo confocal microscopy in primary congenital glaucoma with megalocornea. AB - PURPOSE: To report the microscopic findings of congenital glaucoma-related megalocornea using in vivo confocal microscopy. PATIENTS AND METHODS: Two consecutive adult patients presenting the typical features of glaucomatous megalocornea underwent a complete ophthalmologic examination. The first patient presented with progressive glaucoma with bilateral megalocornea. The second patient's left eye was affected by megalocornea without actual evidence of glaucoma, whereas the right eye was healthy. Both patients were examined using a new-generation scanning slit corneal confocal microscope. RESULTS: In both patients, confocal microscopy revealed a mild reduction of keratocyte density in the mid and rear stroma, a particular abnormal "clew-shaped" morphology of stromal nerves, and the presence of discontinuous hyperreflective structures overhanging the endothelial layer at the level of the Descemet membrane. The endothelium showed severe polymegethism, pleomorphism, and a markedly decreased cell density, and focal cellular lesions were noted. CONCLUSION: Confocal microscopy is a diagnostic tool used to evaluate microscopic aspects of Haab striae and endothelial morphologic changes in glaucomatous megalocornea. Unsuspected alterations, such as nerves abnormalities and focal endothelial tractions by scar tissue, were observed. PMID- 11912355 TI - Intraocular pressure-lowering effects of latanoprost and brimonidine therapy in patients with open-angle glaucoma or ocular hypertension: a randomized observer masked multicenter study. AB - PURPOSE: To compare the effect of treatment with latanoprost or brimonidine on intraocular pressure in patients with glaucoma or ocular hypertension and intraocular pressure inadequately controlled by monotherapy or dual therapy. PATIENTS AND METHODS: Three hundred seventy-nine patients with primary open-angle glaucoma or ocular hypertension were recruited for this 6-month prospective, randomized, observer-masked multicenter study involving 30 eye clinics. All patients were receiving monotherapy or dual therapy that did not adequately control intraocular pressure. After appropriate washout periods, patients were randomized to treatment with latanoprost once daily or brimonidine twice daily. The main outcome measure was change in mean diurnal intraocular pressure after 6 months of treatment compared with baseline. RESULTS: Of the 379 randomized patients, 375 were included in the intent-to-treat analysis. From an overall baseline mean intraocular pressure of 25.0 mm Hg, latanoprost monotherapy reduced mean diurnal intraocular pressure by 7.1 +/- 3.3 mm Hg (mean +/- SD, P < 0.001), whereas brimonidine monotherapy yielded an intraocular-pressure reduction of 5.2 +/- 3.5 mm Hg (P < 0.001). This 1.9 mm Hg difference in intraocular-pressure reduction was significantly in favor of latanoprost (P < 0.001). Ocular allergy (P < 0.001) and systemic side effects (P = 0.005) were reported significantly less frequently by latanoprost-treated patients compared with brimonidine-treated patients. CONCLUSIONS: Both latanoprost and brimonidine reduced intraocular pressure in patients with glaucoma or ocular hypertension after 6 months of treatment. However, latanoprost once daily was significantly more effective than brimonidine twice daily in reducing mean diurnal intraocular pressure. Latanoprost was better tolerated with less frequently occurring ocular allergy and systemic side effects. PMID- 11912356 TI - Discriminant analysis formulas of optic nerve head parameters measured by confocal scanning laser tomography. AB - PURPOSE: To evaluate whether discriminant analysis formulas of optic disc variables measured by confocal laser scanning tomography can detect glaucomatous visual field defects, to compare the diagnostic precision of these formulas to detect glaucomatous visual field defects in different types of chronic open-angle glaucoma, and to assess whether gender or refractive error influence the results obtained by the formulas. METHODS: One hundred and sixty-one patients with perimetrically defined glaucomatous optic nerve damage and 194 healthy subjects were recruited. All patients underwent confocal laser scanning tomography of the optic disc. The data were analyzed with three linear discriminant analysis formulas (sectorial, Bathija, and Mikelberg) obtained in sets of data different from those used in the present study. RESULTS: The areas under the receiver operating characteristic curves of the three formulas and of the cup shape measure as a single parameter ranged from 0.649 to 0.81 in the entire group, and the results did not change when age-matched eyes were considered (0.618-0.812). In each of the glaucoma subgroups with primary open-angle, pseudoexfoliative, and normal-pressure glaucoma, and additionally in the hyperopic, myopic, female, and male subgroups, the sectorial formula had the highest diagnostic precision and the highest correlation coefficients with the visual field indices, followed by the Bathija and Mikelberg formulas, without major differences between the subgroups. All three formulas were more effective than the cup shape measure as a single parameter. CONCLUSION: In the various chronic open-angle glaucomas, the sectorial and Bathija formulas tended to have higher diagnostic precision than the Mikelberg formula and the cup shape measure. Gender and refractive error do not markedly influence the diagnostic precision of the formulas tested. The scores of the formulas are mild indicators of the amount of glaucomatous visual field loss. All three formulas are superior to the single cup shape measure in the detection of glaucomatous optic nerve damage. PMID- 11912357 TI - Endothelin 1 levels in the aqueous humor of dogs with glaucoma. AB - PURPOSE: Endothelin 1 is a small peptide that is involved in regulation of intraocular pressure and modulation of ocular circulation. To investigate the role of endothelin 1 in canine glaucoma, the authors measured aqueous humor levels of endothelin 1 in healthy dogs and in dogs with hypertensive glaucoma. METHODS: Aqueous humor samples were obtained with general anesthesia from the eyes of healthy dogs (n = 5) and dogs with hypertensive glaucoma (n = 10). Measurements were made by enzyme immunoassay for endothelin 1. RESULTS: The endothelin 1 aqueous humor range was 1.12 - 3.63 pg/mL for healthy dogs and 1.97 14.97 pg/mL for glaucomatous dogs. The healthy and glaucomatous canine endothelin 1 aqueous levels (mean +/- SD) were 2.33 +/- 0.90 and 8.11 +/- 5.03 pg/mL, respectively. A two-way analysis of variance indicated that this difference was significant (P = 0.0084). The effect of age on endothelin 1 levels was not significant (P = 0.6283). The large variability found within the glaucomatous group could be explained by the degree of damage of the retina (P = 0.0006). There was no significant correlation between intraocular pressure and endothelin 1 aqueous humor levels within the glaucomatous group (P = 0.29). CONCLUSIONS: The aqueous humor of dogs with hypertensive glaucoma contains significantly higher levels of endothelin 1 than that of healthy dogs. PMID- 11912358 TI - Long-term effects of simultaneous subconjunctival and subscleral mitomycin C application in repeat trabeculectomy. AB - PURPOSE: To assess the efficacy and safety of simultaneous mitomycin C application under conjunctival and scleral flaps in patients with repeat trabeculectomy. METHODS: A total of 44 patients (44 eyes) with previous failed filtering surgery were randomized to one of two groups. The both-flaps group comprised 22 patients (22 eyes) with trabeculectomy and intraoperative mitomycin C application under conjunctival and scleral flaps, whereas the subconjunctival group comprised 22 patients (22 eyes) with subconjunctival application of mitomycin C. Particular attention was paid to intraocular pressure, postoperative medications, visual acuity, filtering bleb appearance, and complications. The mean follow-up time was 38.18 +/- 12.48 months. RESULTS: The mean preoperative intraocular pressure decreased from 39.1 +/- 7.3 mm Hg to the postoperative level of 15.6 +/- 4.8 mm Hg in the both-flaps group (P = 0.014), and from 39.4 +/- 8.4 to 18.7 +/- 5.8 mm Hg in the subconjunctival group (P = 0.018). There was a statistically significant difference in intraocular pressure at all follow-up times, except at 1 week and 1 month postoperatively. Kaplan-Meier survival analysis showed there was no significant difference in total success rate (complete plus qualified success) between the two groups (P = 0.622, log-rank test). However, the two survival curves for the complete success subgroups (without additional medications) confirmed that mitomycin C applications under both flaps had a higher success rate than subconjunctival application (P = 0.043, log-rank test). No statistically significant difference in medications was present between the two groups, and no severe complications developed in either group. CONCLUSIONS: Trabeculectomy augmented with mitomycin C application at both sites could produce a greater lowering of intraocular pressure with low incidence of postoperative complications, and could provide an increased chance of long term success. The procedure is effective and safe in patients with repeat trabeculectomy. PMID- 11912359 TI - Twelve-month evaluation of brimonidine-purite versus brimonidine in patients with glaucoma or ocular hypertension. AB - PURPOSE: To compare the efficacy and safety of brimonidine-Purite (Alphagan; Allergan, Irvine, CA) 0.15% and 0.2% three times daily with brimonidine (Alphagan) 0.2% three times daily in patients with glaucoma or ocular hypertension. PATIENTS AND METHODS: In this 12-month, randomized, multicenter, double-masked, parallel-group study, patients were randomly assigned to receive brimonidine-Purite 0.15% (n = 381), brimonidine-Purite 0.2% (n = 383), or brimonidine 0.2% (n = 383) three times daily. Visits were conducted before the study, at baseline, at weeks 2 and 6, and at months 3, 6, 9, and 12. Diurnal intraocular pressure was measured at 8 am, 10 am, 3 pm, and 5 pm at baseline, week 6, and at months 3, 6, and 12. Intraocular pressure was also measured at 8 and 10 am at week 2 and month 9. Safety was evaluated by adverse events and other ocular and systemic measures. RESULTS: At baseline, mean intraocular pressure was similar in the three treatment groups. During follow-up, there were no statistically significant among-group differences in mean intraocular pressure or mean changes from baseline intraocular pressure (at peak or trough). The difference in mean intraocular pressure between the brimonidine-Purite-0.15% and brimonidine-0.2% treatment group was less than 1 mm Hg at all time points. The relative percent difference in allergic conjunctivitis was 41% lower in the brimonidine-Purite 0.15% group compared with the brimonidine 0.2% group. The comfort and satisfaction rating significantly favored brimonidine-Purite 0.15%. CONCLUSIONS: Over 12-months, brimonidine-Purite 0.15% and 0.2% provided intraocular pressure lowering comparable with brimonidine 0.2% in patients with glaucoma or ocular hypertension. Brimonidine-Purite 0.15% showed the most favorable safety and tolerability profile with a reduced incidence of allergic conjunctivitis and better satisfaction and comfort rating. PMID- 11912360 TI - Effects of tranilast on filtering blebs: a pilot study. AB - PURPOSE: To investigate the effects of topical instillation of 0.5% tranilast eye drops on intraocular pressure (IOP) and bleb formation after glaucoma filtering surgery. PATIENTS AND METHODS: This was a prospective, randomized, double-masked, and controlled clinical trial. A total of 52 eyes of 52 patients were randomly assigned to receive either 0.5% tranilast eye drops (24 eyes) or vehicle saline solution (28 eyes) 4 times daily for 3 months after trabeculectomy. Features of the bleb, such as vascularization and size, and intraocular pressure were studied. RESULTS: The incidence of vascularized bleb and "pseudopod" formation 6 months post treatment were more common in tranilast-treated eyes than control eyes (P = 0.019 and P = 0.043, respectively). The bleb was significantly larger at 6 and 12 months (P = 0.024 and P = 0.049, respectively), and reduction of the IOP was more significant for 2 years postoperatively (P = 0.002 to P = 0.032) in tranilast-treated eyes than control eyes. No vision-threatening side reactions were associated with tranilast. CONCLUSIONS: The use of topical tranilast after filtering surgery alleviates ischemia of the filtering bleb, reduces IOP, and increases the size of the bleb. PMID- 11912361 TI - Timolol concentrations in rat ocular tissues and plasma after topical and intraperitoneal dosing. AB - PURPOSE: Topical beta-blockers, such as timolol, have been used extensively in the medical treatment of glaucoma to lower intraocular pressure (IOP). Recently, these drugs have been shown to have effects on the retinal and optic nerve circulation as well as potential neuroprotective properties. In the current study, the concentration of timolol attained in the cornea, iris-ciliary body, retina, vitreous, and plasma was measured after topical or intraperitoneal administration in rats to determine the relative contributions of each route to intraocular timolol concentrations. MATERIALS AND METHODS: One group of rats received one drop of commercially available 0.5% timolol in the right eye and two drops in the left eye for 3 to 12 days. Another group of rats received one drop of 0.5% timolol in one eye only and concentrations were studied in the ocular tissues at 15, 30, 60, 120, and 240 minutes after instillation. The final group of rats received a single intraperitoneal injection of timolol ranging in concentration from 5 to 75 mg/kg after which tissue and plasma concentrations were measured 30 minutes after injection. All tissue and plasma concentrations were measured by high performance liquid chromatography. RESULTS: Rats that received topical timolol daily for 3 to 12 consecutive days accumulated timolol concentrations of 2.3 to 4.4 microg/g in cornea, 198 to 326 microg/g in iris, 0.05 to 0.11 microg/ml in vitreous, and 0.17 to 0.77 microg/g in retina. In rats that received a single drop of timolol in one eye, the tissue concentrations were higher in the treated eye than in the untreated eye in all cases except for vitreous. In these experiments, timolol levels in plasma were either low or not detectable. Increasing timolol doses administered intraperitoneally resulted in corresponding increased tissue and plasma concentrations. CONCLUSIONS: Absorption of drug into the systemic circulation plays a significant role in delivering timolol to the retina and vitreous in addition to a local ocular route. A clear dose-response relationship exists in all ocular tissues studied after an intraperitoneal dose of timolol. High doses of timolol were required to achieve measurable concentrations of drug in the ocular tissues via our high performance liquid chromatography assay suggesting that a significant hepatic first-pass effect may be involved after an intraperitoneal injection of timolol. PMID- 11912362 TI - A comparison of glaucoma drainage implant tube coverage. AB - PURPOSE: Surgeons may use various materials, including donor sclera, dura, or pericardium grafts to cover glaucoma drainage implant tubes, prior to repositioning conjunctiva. We reviewed our experience with these materials. PATIENTS AND METHODS: Sixty-four eyes with at least 24 months follow-up status post glaucoma drainage implant surgery were evaluated for signs of tube erosion, as well as patch graft thinning, after initial placement of donor sclera (23), dura (18), or pericardium (23) patch grafts. RESULTS: Sixty-two eyes required no intervention for conjunctival and patch graft melting with subsequent tube erosion. Three cases (two eyes) of erosion requiring reoperation (1-dura at 6 months, 1-sclera at 15 months, and in the same eye 21 months later, 1 pericardium) were noted. Significant thinning of the donor patch graft such that the tube was visible beneath intact conjunctiva ocurred in 6 out of 23 donor sclera eyes, 4 out of 18 donor dura eyes, and 6 out of 23 donor pericardium eyes. CONCLUSION: No material was more prone to melting than another. Donor sclera may be slightly more cost-efficient, but gamma-irradiated pericardium has sterility advantages. PMID- 11912363 TI - Comparison of two transducers for color Doppler imaging of the retrobulbar vessels. AB - PURPOSE: To compare blood flow velocity measurements obtained with two different linear probes (7.5 MHz and 12 MHz). MATERIALS AND METHODS: CDI measurements were performed in 31 eyes of 19 individuals with a 12 MHz and a 7.5 MHz probe. Measurements were taken in the ophthalmic artery (OA), central retinal artery (CRA), temporal short posterior ciliary arteries (TSPCA), and nasal short posterior ciliary arteries (NSPCA). With each probe, three measurements of each vessel were obtained, and end-diastolic velocity (EDV), peak systolic velocity (PSV), and resistance index (RI) were calculated. RESULTS: For EDV, the variation among replicates with the 7.5 MHz probe was between 13.99% and 19.51% and with the 12 MHz probe between 15.40% and 23.17%. For PSV, the variation among replicates was between 5.16% and 9.98% and between 6.58% and 10.71%, respectively with each probe. For RI, the variation among the replicates was between 3.10% and 8.97% and between 3.93% and 10.16%, respectively with each probe. The variation between probes was between 15.02% and 23.07% for EDV, between 10.72% and 13.24% for PSV, and between 4.27% and 6.05% for RI. The difference between estimated means was only significant for EDV in the TSPCA (P = 0.019), PSV in the TSPCA (P = 0.0002), as well as NSPCA (P = 0.003) and RI in the CRA (P < 0.001) and NSPCA (P < 0.044). CONCLUSION: The observed variation among three measurements is similar with both the 7.5 MHz and the 12 MHz probes. However, the 7.5 MHz probe tends to measure slightly higher values for some parameters. PMID- 11912364 TI - Glaucoma patients' assessment of their visual function and quality of life. AB - PURPOSE: To determine how glaucoma and glaucoma suspect patients' rating of their vision correlates with Esterman binocular visual field testing and other visual function tests. METHODS: One hundred ninety-one glaucoma patients and 46 glaucoma suspect patients underwent binocular visual field testing and evaluated their vision using the linear rating scale and time-tradeoff utility tests, the National Eye Institute Visual Functional Questionnaire (NEI VFQ-25), and the Short Form 36 (SF-36) quality-of-life instruments. RESULTS: The mean Esterman score was 88.2 +/- 17.4 for the glaucoma subjects and 95.2 + 6.9 for glaucoma suspect subjects (maximum score 100). On a scale from 0 (blind) to 100 (ideal), the mean rating of vision for glaucoma patients and glaucoma suspect patients was 74.8 +/- 17.3 and 78.9 +/- 18.5, respectively. The Esterman test correlated moderately with the overall NEI VFQ-25 score (partial correlation coefficient (PCC) = 0.32, P = 0.001), but only weakly with the linear rating scale (PCC = 0.17, P = 0.02), and the time-tradeoff (PCC = 0.14, P = 0.06). CONCLUSION: Utility values that glaucoma and glaucoma suspect patients assign to their vision do not correlate well with Esterman results. A challenge for the future is the design of clinical tests of vision that better correlate with patient perceptions. PMID- 11912365 TI - Aniridia. PMID- 11912366 TI - Caffeine and intraocular pressure in a Nigerian population. PMID- 11912367 TI - Contemporary management of stage T1 transitional cell carcinoma of the bladder. AB - PURPOSE: Transitional cell carcinoma involving the lamina propria (stage T1) is associated with a high recurrence and progression rate with implications for patient survival and quality of life. A better understanding of the natural history of and treatment alternatives for this tumor may improve the outcome in patients with this stage of bladder cancer. MATERIALS AND METHODS: Literature of the last decade was comprehensively reviewed in regard to clinical and pathological diagnosis, adjuvant treatments, prognosis, and the role and timing of cystectomy. The information was gathered from MEDLINE, current urology journals, abstracts from recent urological meetings and personal experience. RESULTS: High grade and the depth of lamina propria invasion are important prognostic factors. Early diagnosis and accurate pathological assessment are essential for determining the most adequate treatment pathway. Initial treatment consists of complete transurethral resection and adjuvant treatment with intravesical instillation of bacillus Calmette-Guerin (BCG). Immediate postoperative instillation of mitomycin C decreases the risk of recurrence possibly related to tumor implantation. Intravesical treatment does not substantially decrease the chance of progression. Lack of a complete response to BCG at 3 to 6 months, high grade, the depth of lamina propria invasion, the association of carcinoma in situ and prostate mucosa or duct involvement represent significant predictors for progression. Cystectomy should be suggested for recurrent stage T1 tumor after BCG, new onset or persistent carcinoma in situ, tumor located at a difficult site for resection, prostatic duct or stromal involvement and muscle invasion. CONCLUSIONS: High grade stage T1 transitional cell carcinoma is a highly malignant tumor. Complete resection followed by immediate mitomycin C instillation and 6 weekly BCG instillations results in an acceptably low recurrence and progression rate. Rigorous long-term surveillance and continuous reconsideration of radical cystectomy in concordance with the evolution of the disease are essential. PMID- 11912368 TI - Clean, intermittent self-catheterization in the treatment of urinary tract disease. 1972. PMID- 11912369 TI - The uncertainty of radio frequency treatment of renal cell carcinoma: findings at immediate and delayed nephrectomy. AB - PURPOSE: Radio frequency thermal therapy for the ablation of renal cell carcinoma has been reported. Outcomes are usually measured by imaging alone. We have performed ex vivo and in vivo experiments using radio frequency in porcine models in our laboratory. We now report our early experience in the treatment of renal cell carcinoma in patients who underwent post-radio frequency radical or partial nephrectomy. MATERIALS AND METHODS: We treated 10 patients diagnosed with small renal masses with radio frequency. All masses were biopsied before treatment. In 4 patients 5 renal cell carcinomas were treated with radio frequency after surgical exposure of the tumor followed immediately by partial or radical nephrectomy (acute group). Six other patients were treated percutaneously with ultrasound or computerized tomography guided radio frequency under local anesthesia and intravenous sedation 7 days before partial or radical nephrectomy (delayed group). A median of 2 radio frequency cycles was applied. Mean total heating time was 17 minutes 15 seconds. Specimens were analyzed grossly and histologically. Triphasic contrast-enhanced computerized tomography and/or magnetic resonance imaging was performed before and 7 days after radio frequency treatment in the delayed group. RESULTS: Mean radiological largest diameter of all 11 masses was 2.4 cm. and mean gross diameter was 2.2 cm. Pathological examination demonstrated residual viable tumor in approximately 5% of the volume in 4 of the 5 tumors in the acute group and in 3 of the 6 masses of the delayed group. In 1 delayed case the viable tumor appeared to be in contact with the renal vein. No significant complications were observed in 9 of the 10 patients. In 1 delayed case, a subcapsular hepatic hematoma, biliary fistula and pneumonia developed and resolved. CONCLUSIONS: Based on our experience, we continue to consider percutaneous radio frequency for the treatment of small renal cell carcinomas as a potentially curative therapy. However, complete tumor cell death appears to be difficult to achieve with our current treatment protocol. More phase II testing is indicated to ensure that this technique is an effective and reproducible treatment alternative. PMID- 11912370 TI - Etiology of spontaneous perirenal hemorrhage: a meta-analysis. AB - PURPOSE: We determine the most common etiology of spontaneous perirenal hemorrhage. MATERIALS AND METHODS: A MEDLINE search of the English language literature from 1985 to 1999 revealed 47 publications and 165 cases of spontaneous renal hemorrhage meeting our study entry criteria. These criteria were presentation of raw data including imaging modality, pathological confirmation (123 cases) or long-term (greater than 2 years) (42) imaging and/or clinical followup and no history of recent trauma, anticoagulant use, dialysis or renal transplant. Meta-analysis was performed using analysis of counts derived from contingency tables and pooled and stratified analysis. RESULTS: Hemorrhage was identified by ultrasound in 56 of 100 cases (56%) and by computerized tomography (CT) in all 135 cases assessed (100%). Etiology was correctly identified with an overall sensitivity and specificity of 0.11 and 0.33 for ultrasound and 0.57 and 0.82 for CT. Angiography in 81 cases revealed active bleeding in 11. The most common etiology of spontaneous renal hemorrhage was benign or malignant neoplasm (101 cases, 61%) with angiomyolipoma being predominant (48) followed closely by renal cell carcinoma (43). Vascular disease was the next most common offender (28 cases, 17%) with polyarteritis nodosa occurring most frequently (20). CONCLUSIONS: The most common cause of spontaneous perirenal hemorrhage is renal neoplasm and approximately 50% of such neoplasms are malignant. CT is the method of choice for evaluation of perirenal hemorrhage, although its sensitivity for detection of underlying etiology is only moderate. PMID- 11912371 TI - A comparison of unenhanced helical computerized tomography findings and renal obstruction determined by furosemide 99m technetium mercaptoacetyltriglycine diuretic scintirenography for patients with acute renal colic. AB - PURPOSE: We assessed unenhanced helical computerized tomography (CT) secondary findings as predictors of renal obstruction as determined by diuretic scintirenography, and determined their reproducibility. MATERIALS AND METHODS: We performed a retrospective review of the records of 77 consecutive patients with unenhanced helical CT findings (stones and secondary findings, including renal parenchymal edema, hydronephrosis, hydroureter, perinephric fat stranding, periureteral fat stranding and extravasation) of urinary lithiasis who had also undergone concomitant diuretic scintirenography during the initial emergency room evaluation during a 1-year period. Unenhanced helical CT films were independently reviewed by 2 attending radiologists (blinded to clinical outcome) to determine interobserver variability. The results were compared to those of diuretic scintirenography. RESULTS: Considerable interobserver variability, as evidenced by kappa values ranging from 0.26 to 0.60, existed for the diagnosis of secondary findings associated with urinary lithiasis on unenhanced helical CT. There was no significant difference in terms of CT findings between patients diagnosed by diuretic scintirenography as having high grade/complete obstruction and those with partial obstruction (p values 0.24 to 0.85 for the 6 unenhanced helical CT findings analyzed). Analyses of variance followed by Tukey's pairwise comparisons showed no significant difference in average number of unenhanced helical CT findings between patients with high grade/complete obstruction (mean plus or minus standard deviation 4.4 +/- 1.31), partial obstruction (4.4 +/- 1.30), and decompression/no obstruction (4.2 +/- 1.16). However, the mean number of unenhanced helical CT findings for patients with normal scintirenography/no obstruction (1.9 +/- 1.41) was significantly different from each of the other 3 diuretic scintirenography groups. Separate logistic regression analyses showed that each unenhanced helical CT finding, except for renal parenchymal edema and urinary extravasation, was a significant predictor of "any degree of obstruction" (high grade and partial obstruction groups) compared to "no obstruction" (decompressed and no obstruction groups). Odds ratios (95% confidence interval) ranged from 6.15 (2.25, 16.82) for perinephric fat stranding to 3.41 (1.30, 8.97) for hydroureter. When these analyses were repeated after exclusion of 8 patients with bladder/passed stones, only perinephric fat stranding and periureteral fat stranding remained significant predictors of "any degree of obstruction," with respective odds ratios of 4.21 (1.49, 11.91) and 4.08 (1.31, 12.65). CONCLUSIONS: Measures of agreement between trained, independent radiologists with respect to unenhanced helical CT secondary findings show considerable variability. The average number of CT consensus findings is not helpful in differentiating patients with variable degrees of obstruction, except for those with normal scintirenography/no obstruction. Unenhanced helical CT findings, except for renal parenchymal edema and urinary extravasation, are significant predictors of "any degree of obstruction" (high grade or partial obstruction) compared to "no obstruction" (decompressed or no obstruction). Therefore, unenhanced helical CT findings may be useful for identifying patients with any degree of obstruction but do not differentiate between those with high grade and partial obstruction. PMID- 11912372 TI - Robotic assisted kidney transplantation: an initial experience. AB - PURPOSE: The use of robotics is a recent innovation in surgery. In addition to dexterity enhancement and motion scaling, this new technology opens the horizon of remote surgery. This latter advancement has potential use during surgery involving a high risk of patient-to-professional or professional-to-patient virus transmission. We investigated the feasibility of robotic assisted kidney transplantation. MATERIALS AND METHODS: A right cadaveric kidney was transplanted into a 26-year-old male patient who has been on hemodialysis for 11 years. Surgery was done with the help of the da Vinci robot (Intuitive Surgical, Inc., Mountain View, California) by a remote surgeon, who completely performed vascular dissection and anastomosis as well as ureterovesical anastomosis. The role of the assistant by the side of the patient was limited to access creation, exposure, hemostasis and maintaining traction on the running sutures performed by the robot. RESULTS: Operative time was 178 minutes. Robotic assistance made anastomosis possible by its unique ability of stereoscopic magnification and ultra-precise suturing techniques due to the flexibility of the robotic wristed instruments. Renal perfusion was excellent with immediate diuresis. Postoperative acute tubular necrosis started to resolve after 1 week. CONCLUSIONS: This study demonstrates that robotic assisted kidney transplantation is feasible. Currently technical and cost hindrances limit the routine use of robots. However, with ongoing improvement and future availability of this technology the prevention of patient-to-professional and professional-to-patient viral transmission may become a potential field of application. PMID- 11912373 TI - A single 24-hour urine collection is inadequate for the medical evaluation of nephrolithiasis. AB - PURPOSE: We determined the adequacy of a single 24-hour urine sample for evaluating patients for medical renal stone prevention. MATERIALS AND METHODS: A total of 459 patients from a private urology practice specializing in the treatment of urolithiasis and 683 from a university stone research clinic provided 2 and 3, 24-hour urine samples, respectively. We used samples 1 and 2 from private practice patients, and 1 and 3 from university clinic patients for analysis, and compared each to the others by correlation coefficients and calculation of the mean difference plus or minus standard deviation (SD) of the difference. Urine risk factors were measured by standard methods. RESULTS: Although the correlation of urine values 1 and 2 was excellent for all stone risk factors, SD values for the differences were large enough that within 1 SD on either side of 0, which included 68.8% of cases, by chance urine 1 would depart from urine 2 by clinically important amounts. These departures would be more than sufficient to misdiagnose common metabolic disorders. CONCLUSIONS: A single 24 hour sample is not sufficient for evaluating patients before metabolic treatment for stone prevention because misdiagnosis is common, leading to inappropriate treatment. PMID- 11912374 TI - Analgesia for shock wave lithotripsy. AB - PURPOSE: We evaluated the effectiveness of and patient preference for analgesia used during shock wave lithotripsy by comparing diclofenac alone with a combination of diclofenac and patient controlled analgesia, that is alfentanil. MATERIALS AND METHODS: A total of 64 patients were treated using a Lithotriptor S (Dornier Medical Systems, Marietta, Georgia) and randomized to receive diclofenac alone or combined with an alfentanil patient controlled analgesia pump. If treated twice, they crossed over to the alternative form of analgesia. A record was maintained of the site and size of the stone, maximum power achieved, number of shocks, amount of alfentanil used and need for additional analgesia. After treatment patients scored on a visual analog scale the maximum level of pain and satisfaction with analgesia. RESULTS: There was no difference in the mean size of the stone treated (8.6 and 7.5 mm.), energy level (71% and 71% or approximately 17 kV.) or number of shocks (3,000 and 2,900, respectively) in the groups. Only 2 patients in the diclofenac group required additional analgesia and there were no significant side effects from either treatment. The mean pain scores were not significantly different in the diclofenac and patient controlled analgesia groups (3.54 and 2.93, respectively, (p = 0.34), although those on patient controlled analgesia were more satisfied (7.72 versus 9.14, p = 0.04). Of the 38 patients who presented twice 58% preferred diclofenac alone. CONCLUSIONS: This study suggests that there is no significant difference in the level of pain experienced with diclofenac alone or when combined with an alfentanil patient controlled analgesia pump during shock wave lithotripsy. However, patients are more satisfied with treatment when a patient controlled analgesia pump is available. PMID- 11912375 TI - Neoinfundibulotomy for the management of symptomatic caliceal diverticula. AB - PURPOSE: Direct percutaneous access provides effective treatment for complex caliceal diverticula. Yet, access into the diverticulum alone is usually tenuous and passage of a guide wire across a stenotic infundibulum is often impossible. An alternative technique is described which creates a "neoinfundibulum" to assist in the management of symptomatic caliceal diverticula. MATERIALS AND METHODS: During a 6-year period 22 patients with symptomatic caliceal diverticula were treated via a percutaneous approach, of whom 21 had calculi within the diverticula. After accessing the diverticulum directly, it was impossible to pass a guide wire through the stenotic infundibulum in 18 (82%) patients, prompting advancement of the access needle through the diverticular wall into the renal pelvis. Once secure access was established, balloon dilation was performed to 30Fr to create the "neoinfundibulum." Percutaneous ultrasonic lithotripsy was performed in the usual fashion. A 22Fr Councill catheter was placed to keep the infundibular tract open for 5 to 7 days to allow complete epithelialization and drainage. Stone-free, symptom-free and complication rates were assessed. RESULTS: Pain, recurrent urinary tract infections and hematuria were the presenting complaints in the subgroup of patients undergoing "neoinfundibulotomy." Average stone burden was 11.7 x 12 mm. and average hospital stay was 2.8 days. Of the patients 94% were symptom-free at 6-week followup, and 80% were stone-free on followup excretory urography. The remaining patients had residual stone fragments less than 3 mm. in diameter. Complications related to access were identified in 2 patients who sustained a pneumothorax after a supra-11th rib access, which was successfully managed with tube thoracostomy. CONCLUSIONS: Percutaneous management of complex caliceal diverticula provides a safe and effective option for symptomatic patients. When the stenotic infundibulum cannot be traversed with a guide wire, creation of a new infundibulum offers a secure alternative for accessing the collecting system, while providing equally effective results. PMID- 11912376 TI - Management of ureteral calculi: a cost comparison and decision making analysis. AB - PURPOSE: We compared the cost of treatment strategies for ureteral calculi using a decision tree model. MATERIALS AND METHODS: A comprehensive literature review was performed to determine the average success rate of each of 3 treatment modalities, namely observation, ureteroscopy and shock wave lithotripsy. Using these success rates decision analysis models were constructed using Data 3.5 software (TreeAge Software, Inc., Williamstown, Massachusetts) to estimate the cost of treatment and followup for each of the 3 treatments. One-way sensitivity analysis was performed to evaluate the effect of varying individual probabilities of success and costs, and 2-way sensitivity analysis was done to evaluate the model for a wide range of potential costs and success rates of ureteroscopy and shock wave lithotripsy. In addition, a table was constructed to enable individual surgeons and institutions to determine the cost impact of ureteroscopy and shock wave lithotripsy in their unique clinical scenarios. RESULTS: Observation was the least costly pathway if no financial cost, such as emergency room visits, was incurred by failed observation. Ureteroscopy was less costly than shock wave lithotripsy for stones at all ureteral locations. A cost difference between the 2 modalities of approximately $1,440, $1,670 and $1,750 was noted for proximal, mid and distal ureteral calculi, respectively. One-way sensitivity analysis showed that the cost of ureteroscopy would have to increase by more than $1,400, $1,700 and $1,850, and the success rate would have to decrease by 28%, 36% and 39% for proximal, mid and distal stones, respectively, before reaching cost equivalence with shock wave lithotripsy. Likewise, the cost of shock wave lithotripsy would have to decrease by more than $1,489 to achieve cost equivalence with ureteroscopy. Overall ureteroscopy was more cost-effective at all stone sites regardless of the success rate of shock wave lithotripsy. CONCLUSIONS: Ureteroscopy is the most cost-effective treatment strategy for ureteral stones at all locations after observation fails. The high cost of purchasing and maintaining a lithotriptor is responsible for the high treatment cost associated with shock wave lithotripsy. However, cost is only one of a number of important factors that are considered when determining an appropriate treatment strategy. PMID- 11912377 TI - Outcome of surgical treatment of isolated local recurrence after radical nephrectomy for renal cell carcinoma. AB - PURPOSE: Isolated local recurrences after radical nephrectomy for renal cell carcinoma occur in 2% to 3% of cases. Today local recurrences can be detected at an early stage due to modern imaging techniques. It remains controversial whether an aggressive surgical approach to this problem can prolong survival. MATERIALS AND METHODS: We retrospectively analyzed 16 patients who were treated surgically at our institution for suspected isolated local renal cell carcinoma recurrence during the last 10 years. All patients had undergone extensive staging and had no evidence of distant metastases with the local recurrence. Surgical exploration confirmed carcinoma recurrence in 13 of the 16 cases and all 13 patients underwent complete resection of the local recurrence. Three patients were found to have had false-positive computerized tomography findings on surgical exploration. RESULTS: Mean time to recurrence was 45.5 months (range 7 to 224). Only 2 patients were symptomatic, while in 11 disease had been detected at routine followup. Mean size of the recurrent tumor was 5.92 cm. (range 2 to 10). All patients survived surgery without major complications. Of the patients 7 died of metastatic disease after a mean survival of 23.1 months (range 4 to 68) following recurrence removal and 6 are alive with a mean survival of 53.0 months (range 18 to 101) (p = 0.09). Time to recurrence after nephrectomy was significantly longer (p <0.05) and size of recurrence significantly smaller (p <0.04) in the patients still alive. In 1 surviving patient evidence of metastatic disease developed 9 months after surgery for recurrence. CONCLUSIONS: Careful followup after radical nephrectomy for renal cell carcinoma allows the diagnosis of small local recurrences before they become symptomatic in the majority of cases. Although most of these patients will eventually have and die of metastatic disease, an aggressive surgical approach is justified and can result in prolonged survival. PMID- 11912378 TI - Stage Ta-T1 bladder cancer: the relationship between findings at first followup cystoscopy and subsequent recurrence and progression. AB - PURPOSE: We studied the relationship of first cystoscopy findings with recurrence and progression rates in a large, population based series of patients with bladder cancer. MATERIALS AND METHODS: All 463 patients with an initial diagnosis of stage Ta-T1 bladder cancer in western Sweden in 1987 to 1988 were followed at least 5 years. The 355 patients who were treated with transurethral resection only until repeat cystoscopy or longer were selected for this report. RESULTS: Negative first cystoscopy findings were associated with significantly decreased recurrence and progression rates for all grades, and for stage Ta and T1 tumors. However, some patients with initial high grade carcinoma (WHO 2 to 3) had stage progression despite negative first cystoscopy. On multivariate analyses first cystoscopy findings and papillary urothelial neoplasm of low malignant potential versus grades 1 to 3 but not stage and the number of tumors had prognostic significance for time to recurrence. Only first cystoscopy findings and grade had prognostic significance for time to stage progression. CONCLUSIONS: Our data support other groups who recommend a less intense cystoscopy followup schedule in patients with negative cystoscopy findings 3 months after initial transurethral bladder resection. We recommend that patients with initial papillary urothelial neoplasm of low malignant potential and low grade carcinoma (WHO 1) with negative first cystoscopy findings undergo repeat cystoscopy at month 12. In our opinion followup should not be less intense in patients with high grade carcinoma (WHO 2 3), even in those with stage pTa disease. PMID- 11912379 TI - Contemporary morbidity from lymphadenectomy for penile squamous cell carcinoma: the M.D. Anderson Cancer Center Experience. AB - PURPOSE: Inguinal lymphadenectomy can be curative in patients with small volume inguinal metastases and those with more significant adenopathy responding to combination chemotherapy. However, several series collected for 15 to 40 years attest to the significant morbidity associated with lymphadenectomy. We reviewed our recent experience with lymphadenectomy in patients with invasive penile cancer who were judged to require inguinal staging and therapeutic procedures to assess the incidence and magnitude of complications caused by this procedure, especially in those with no palpable adenopathy (prophylactic group). MATERIALS AND METHODS: A total of 106 lymphadenectomy procedures were performed in 53 patients. The indications for dissection were prophylactic in 66 (62%) patients in whom a superficial dissection alone was completed on the ipsilateral side, therapeutic in 28 (26%) in whom superficial, deep and ipsilateral pelvic dissections were performed, and palliative in 12 (11%) undergoing extensive resection of inguinal and abdominal wall tissue after chemotherapy. Minor postoperative complications included those requiring local wound debridement in the clinic, mild to moderate leg edema, seroma formation not requiring aspiration and minimal skin edge necrosis requiring no therapy. Major complications included severe leg edema interfering with ambulation, skin flap necrosis requiring a skin graft, rehospitalization, deep venous thrombosis, death, or reexploration or other invasive procedures performed in the operating room. The incidence and magnitude of complications were compared with prior reports from our center and other series. RESULTS: A total of 41 (68%) minor and 19 (32%) major complications occurred with the 106 dissections (31 of 53 patients, 58%). Prophylactic and therapeutic dissections were associated with a lower incidence of complications compared with palliative dissections (p = 0.017 to 0.049). The incidence of major complications also trended lower in the prophylactic group compared with other indications (p = 0.05). One patient in the palliative group died of sepsis on postoperative day 15. When compared with 3 prior series, the incidence of skin edge necrosis in our series was significantly lower (8% versus 45% to 62%, p <0.0001). Similarly, the incidence and severity of edema in our series were significantly lower than in a prior report from our institution (23% versus 50%, p <0.0001). CONCLUSIONS: For select patients undergoing prophylactic inguinal dissection to detect the presence of microscopic metastases, the incidence and magnitude of complications appeared acceptable in our contemporary experience. Similarly the morbidity of therapeutic lymphadenectomy appeared acceptable, considering the potential therapeutic benefit. However, significant complications, including death, can be associated with palliative groin dissection. Optimal candidates are those having a significant response to systemic chemotherapy whose groins are grossly uninfected. PMID- 11912380 TI - Dendritic cell infiltration in a patient with seminomatous germ cell tumor of the testis: is there a relationship with infertility and tumor stage? AB - PURPOSE: Inflammatory cells, such as dendritic cells, are considered to trigger the antitumoral immune response against tumors, such as testicular cancer. Male infertility associated with cancer may be due to endocrine or immunological factors. We investigated possible associations of antigen expression with dendritic cells, histiocytic cells and seminoma stage as well as with impaired spermatogenesis. MATERIALS AND METHODS: From 1992 to 1999, 30 patients with seminoma underwent orchiectomy at our center, including 14 who underwent spermiography before orchiectomy. Streptavidin-biotin immunostaining was performed on paraffin -embedded tumor specimens using antibodies against protein S-100 for dendritic cells and CD68-KP1 antigen. RESULTS: Light infiltration by less than 20 dendritic cells and less than 103 CD68+ cells was associated with tumor size greater than 1.5 cm. in 75% and 80% of patients, respectively. Strong infiltration by greater than 20 dendritic cells and greater than 103 CD68+ cells was associated with negative lymph nodes in 86% of patients. Slight infiltration by dendritic cells was observed in 71% of patients with a sperm count of greater than 8.6 million per ml. and in 100% with more than 45% motile sperm (p not significant and 0.02, respectively). Necrospermia increased with dendritic and CD68+ cell infiltration. No association was noted among preoperative serum tumor marker levels, the sperm count and immunostaining. CONCLUSIONS: Sperm autoimmunity is a plausible mechanism of infertility in men with germ cell tumors. Dendritic cells may induce antitumor cell cytotoxic reactions, but may also be cytotoxic to sperm cells or lead to inhibited spermatogenesis. Further studies focusing on tumor rejection antigen and the cloning of specific cytotoxic T lymphocyte against gametes are required to confirm these finding. PMID- 11912381 TI - Comparison of contrast enhanced color Doppler targeted biopsy with conventional systematic biopsy: impact on prostate cancer detection. AB - PURPOSE: We performed a prospective study to determine whether a limited biopsy approach with contrast enhanced color Doppler ultrasound targeted biopsy of the prostate would detect cancer as well as gray scale US guided systematic biopsy with a larger number of biopsy cores. MATERIALS AND METHODS: We examined 230 male screening volunteers with a total prostate specific antigen of 1.25 ng./ml. or greater and free-to-total prostate specific antigen less than 18%. Two independent examiners evaluated each subject and a single investigator performed 5 or fewer contrast enhanced targeted biopsies into hypervascular regions in the peripheral zone during intravenous infusion of the US contrast agent Levovist (Schering, Berlin, Germany). Subsequently another examiner performed 10 systematic prostate biopsies. The cancer detection rates of the 2 techniques were compared. RESULTS: Cancer was detected in 69 of the 230 patients (30%), including 56 (24.4%) by contrast enhanced targeted biopsy and in 52 (22.6%) by systematic biopsy. Cancer was detected by targeted biopsy alone in 17 patients (7.4%) and by systematic biopsy alone in 13 (5.6%). The overall cancer detection rate by patient was not significantly different for targeted and systematic biopsy (p = 0.58). The detection rate for targeted biopsy cores (10.4% or 118 of 1,139 cores) was significantly better than for systematic biopsy cores (5.3% or 123 of 2,300 cores, p <0.001). Contrast enhanced targeted biopsy in a patient with cancer was 2.6-fold more likely to detect prostate cancer than systematic US guided biopsy. CONCLUSIONS: Contrast enhanced color Doppler targeted biopsy detected as many cancers as systematic biopsy with fewer than half the number of biopsy cores. Although an increase in cancer detection was achieved by combining targeted and systematic techniques in this screening population, contrast enhanced targeted biopsy alone is a reasonable approach for decreasing the number of biopsy cores. PMID- 11912382 TI - How well does the Partin nomogram predict pathological stage after radical prostatectomy in a community based population? Results of the cancer of the prostate strategic urological research endeavor. AB - PURPOSE: The Partin nomogram uses preoperative Gleason grade, serum prostate specific antigen and clinical stage to predict pathological outcome after radical prostatectomy. It was developed and validated in a select population of patients at 3 academic institutions. Although the nomogram is widely used, it has yet to be validated in a community based population. We assessed the performance of the nomogram in the Cancer of the Prostate Strategic Urological Research Endeavor, a nationwide, community based observational disease registry of men with prostate cancer. MATERIALS AND METHODS: Included in the cohort were 1,162 men in Cancer of the Prostate Strategic Urological Research Endeavor who underwent radical prostatectomy. Using various probability thresholds the nomogram was used to predict outcomes in each patient. Using different probability thresholds receiver operating characteristics curves were then used to assess test performance. RESULTS: Of the men 860 (74%) had organ confined disease, 179 (15%) had established capsular penetration, 95 (8%) had seminal vesicle involvement and 37 (3%) had lymph node involvement. Calculated receiver operating characteristics curve area was 0.684 for predicting organ confined disease, 0.614 for predicting capsular penetration, 0.726 for predicting seminal vesicle involvement and 0.766 for predicting lymph node involvement. These values were lower than in previously published reports. CONCLUSIONS: While the Partin nomogram performs adequately in a community based cohort of men who undergo radical prostatectomy for localized prostate cancer, it does not attain the success demonstrated in previous studies in select academic cohorts. This result was likely due to differences in the distribution of pathological outcomes in the community based cohort, in which more men had organ confined disease. PMID- 11912383 TI - Early onset prostate cancer: predictors of clinical grade. AB - PURPOSE: Prostate cancer is typically a disease of elderly men and, therefore, it has not been well characterized in those affected at a young age. With the advent of serum prostate specific antigen testing, we are able to detect prostate cancer in young men even in the absence of symptoms. We studied a large group of early onset prostate cancer cases to illustrate the clinical presentation of men with early onset prostate cancer and to determine the effect of family history on Gleason grade as a reflection of prognosis. MATERIALS AND METHODS: All study participants were enrolled in the University of Michigan Prostate Cancer Genetics Project. Enrollment criterion of the Prostate Cancer Genetics Project includes a diagnosis of prostate cancer at age 55 years or younger. Descriptive statistics and logistic regression were used to characterize early onset prostate cancer and assess the associated prognostic factors. RESULTS: The study group was comprised of 257 men with prostate cancer diagnosed at age 55 years or younger. Median age at diagnosis was 51 years (range 34 to 55) and almost half of the participants reported a negative family history of prostate cancer. In logistic regression analysis having an affected father, an affected first-degree relative or an affected relative of any relation was each a statistically significant predictor of well differentiated (Gleason 6 or less) compared to moderately and poorly differentiated prostate cancer (Gleason 7-10) after adjusting for confounding variables. Men with an affected relative were nearly twice as likely to have well differentiated prostate cancer compared to men without affected relatives. CONCLUSIONS: Family history appears to predict the development of well differentiated tumors independently. In our study men with no family history of prostate cancer had higher grade tumors, which are associated with a more serious prognosis. Future studies of early onset prostate cancer should be directed toward identifying additional risk factors that may be relevant for men without a family history of the disease. PMID- 11912384 TI - Feasibility study: watchful waiting for localized low to intermediate grade prostate carcinoma with selective delayed intervention based on prostate specific antigen, histological and/or clinical progression. AB - PURPOSE: We assessed the feasibility of a watchful waiting protocol with selective delayed intervention using clinical, prostate specific antigen (PSA) or histological progression as treatment indications for clinically localized prostate cancer. MATERIALS AND METHODS: In this prospective, single arm cohort study patients with favorable clinical parameters (stage T1b to T2b N0M0, Gleason score 7 or less and PSA 15 ng./ml. or less) are conservatively treated with watchful waiting. When a patient meets disease progression criteria, arbitrarily defined by the 3 parameters of the rate of PSA increase, clinical progression or histological upgrade on repeat prostate biopsy, appropriate treatment is implemented. Patients are followed every 3 months for the first 2 years and every 6 months thereafter. Serum PSA measurement and digital rectal examination are done at each visit and repeat prostate biopsy is performed 18 months after study enrollment. RESULTS: Since November 1995, the study has accrued 206 patients with a median followup of 29 months (range 2 to 66). Of these men 137 remain on the surveillance protocol with no disease progression, while 69 were withdrawn from study for various reasons. There was clinical, PSA and histological progression in 16, 15 and 5 cases, respectively. The estimated actuarial probability of remaining on the surveillance protocol was 67% at 2 years and 48% at 4. The probability of remaining progression-free was 81% and 67% at 2 and 4 years, respectively. CONCLUSIONS: A policy of watchful waiting with selectively delayed intervention based on predefined criteria of disease progression is feasible. This strategy offers the benefit of an individualized approach based on the demonstrated risk of clinical or biochemical progression with time and, thus, it may decrease the burden of therapy in patients with indolent disease, while providing definitive therapy for those with biologically active disease. PMID- 11912385 TI - A phase 3, multicenter, open label, randomized study of abarelix versus leuprolide plus daily antiandrogen in men with prostate cancer. AB - PURPOSE: We compared the endocrinological and biochemical efficacy of abarelix depot, a gonadotropin-releasing hormone antagonist, with that of a widely used combination of luteinizing hormone releasing hormone agonist and a nonsteroidal antiandrogen. MATERIALS AND METHODS: A total of 255 patients were randomized to receive open label 100 mg. abarelix depot or 7.5 mg. leuprolide acetate intramuscularly injection on days 1, 29, 57, 85, 113 and 141 for 24 weeks. Patients in the abarelix group received an additional injection on day 15 and those in the leuprolide acetate group received 50 mg. bicalutamide daily. Patients could continue treatment with study drug for an additional 28 weeks. The efficacy end points were the comparative rates of avoidance of testosterone surge (greater than 10% increase) within 7 days of the first injection and the rapidity of achieving reduction of serum testosterone to castrate levels (50 ng./dl. or less) on day 8. Patients were monitored for adverse events and laboratory abnormalities. RESULTS: Abarelix was more effective in avoidance of testosterone surge (p <0.001) and the rapidity of reduction of testosterone to castrate levels on day 8 (p <0.001) than combination therapy. No significant difference was seen between the groups in the initial rate of decline of serum prostate specific antigen or the ability to achieve and maintain castrate levels of testosterone. No unusual or unexpected adverse events were reported. CONCLUSIONS: Abarelix as monotherapy achieves medical castration significantly more rapidly than combination therapy and avoids the testosterone surge characteristic of agonist therapy. Both treatments were equally effective in reducing serum prostate specific antigen, and achieving and maintaining castrate levels of testosterone. PMID- 11912386 TI - Outcome of patients with Gleason score 8 or higher prostate cancer following radical prostatectomy alone. AB - PURPOSE: We determine the effect of clinical and pathological variables on the outcome of patients with prostate cancer of Gleason scores 8 or greater treated with radical prostatectomy alone. MATERIALS AND METHODS: Between April 1987 and October 1998, 1,199 patients underwent radical retropubic prostatectomy. We identified 188 patients assigned a Gleason score of 8 or higher in the prostatectomy specimen who did not receive any neoadjuvant or adjuvant therapy. Median followup was 60 months (range 1 to 129). Disease recurrence was defined as any detectable prostate specific antigen level 0.1 ng./ml. or greater. RESULTS: Of 188 patients 128 (68%) had no evidence of prostate cancer after a median followup of 60 months, while 60 (32%) demonstrated a detectable PSA level. There were 58 (31%) patients with disease confined to the prostate with negative surgical margins while 108 (57%) had prostate cancer confined to the surgical specimen. Positive surgical margin with extraprostatic extension was seen in 16 (9%) patients and seminal vesicle invasion was present in 40 (21%). The 5 and 7 year disease-free survival rates for the entire cohort were 71% and 55%, respectively. Patients with specimen confined disease had a significantly higher 5-year disease-free survival rate than those with nonspecimen confined disease (84% and 50%, p <0.0001). On multivariate analysis pathological status of the surgical specimen was the most significant independent predictor of disease recurrence. Age, ethnicity, clinical stage and preoperative PSA had no independent effect on disease recurrence. CONCLUSIONS: Long-term disease-free survival can be expected in those patients with high grade prostate cancer whose disease is confined to the prostate and/or the surgical specimen. PMID- 11912387 TI - Extended pelvic lymphadenectomy in patients undergoing radical prostatectomy: high incidence of lymph node metastasis. AB - PURPOSE: Lymphadenectomy for prostate cancer is limited to obturator and external iliac lymph nodes, although the internal lymph nodes represent the primary landing zone of lymphatic drainage. We performed anatomically adequate extended pelvic lymphadenectomy to assess the incidence of lymph node metastasis in cases of clinically localized prostate cancer. MATERIALS AND METHODS: A total of 103 consecutive patients underwent extended pelvic lymphadenectomy at radical retropubic prostatectomy comprising 9 selective fields, namely the external iliac, internal iliac, obturator and common iliac lymph nodes bilaterally, and the presacral lymph nodes. Histopathological findings were compared with serum prostate specific antigen (PSA), histopathological stage, preoperative biopsy and postoperative prostatectomy Gleason score. Extended pelvic lymphadenectomy was compared with radical retropubic prostatectomy and standard lymphadenectomy in 100 consecutive patients in terms of complications, the number of lymph nodes dissected and operative time. RESULTS: There were no significant differences in age, preoperative PSA or mean biopsy Gleason score in patients who underwent extended pelvic and standard lymphadenectomy. Metastases were diagnosed in 27 of the 103 patients (26.2%) who underwent the extended procedure. A mean of 28 lymph nodes (range 21 to 42) were dissected. Metastases were identified in the internal iliac and presacral regions despite negative obturator lymph nodes. Of the 27 patients 1 to 3 lymph nodes involved with metastasis were detected in 15, 9 and 1, respectively. In 26 of the 27 patients (95.8%) with lymph node metastasis PSA was greater than 10.5 ng./ml. and preoperative biopsy Gleason sum was 7 or greater. A low risk of 2% for lymph node disease was noted in patients with serum PSA less than 10.5 ng./ml. and biopsy Gleason sum less than 7. There were no significant differences in regard to intraoperative and postoperative complications, lymphocele formation or blood loss in the 2 groups. CONCLUSIONS: Extended pelvic lymphadenectomy is associated with a high rate of lymph node metastasis outside of the fields of standard lymphadenectomy in cases of clinically localized prostate cancer. Lymphadenectomy including the internal iliac lymph nodes should be performed in all patients with prostate cancer who are at high risk for lymph node involvement, as indicated by PSA greater than 10.5 ng./ml. and biopsy Gleason sum 7 or greater. In the low risk group pelvic lymphadenectomy can be omitted. PMID- 11912388 TI - Low dose unenhanced helical computerized tomography for the evaluation of acute flank pain. AB - PURPOSE: Unenhanced helical computerized tomography (CT) has proved to be an excellent diagnostic tool for evaluating acute flank pain with reported 95% to 100% sensitivity, 92% to 100% specificity, 96% to 100% positive and 91% to 100% negative predictive values. The diagnostic value of a new low dose protocol was prospectively studied and compared with the results of conventional unenhanced helical CT in a previous series with an effective dose equivalent (HE) of 3.1 to 4.3 mSv. and in current literature with an estimated HE of 4.3 to 4.7 mSv. MATERIALS AND METHODS: In 109 patients 18 to 86 years old with acute flank pain we performed low dose unenhanced helical CT in addition to abdominal ultrasound and urinalysis with new CT parameters (120 kV. 70 mA., 5 mm. collimation, pitch 2 and incremental reconstruction each 5 mm.) that led to a more than 50% decrease in radiation exposure to 1.50 mSv. in females and 0.98 mSv. in males. Ureteral calculi were confirmed or excluded by retrograde ureteropyelography in 51 cases. In the other cases the diagnosis was verified by the clinical and ultrasound course, and/or stone asservation. RESULTS: In 80 of the 109 patients the flank pain was caused by a ureteral calculus. Low dose unenhanced helical CT precisely identified 77 ureteral calculi with 1 false-positive finding. Thus, the sensitivity and specificity of low dose unenhanced helical CT were 96% and 97% with a 99% positive and 90% negative predictive value. In 15 of 29 patients with CT findings negative for stone disease different causes of pain were established by low dose unenhanced helical CT. CONCLUSIONS: Even with the significantly decreased radiation exposure of the low dose protocol unenhanced helical CT is still an excellent and rapid diagnostic tool for evaluating acute flank pain with lower radiation exposure than excretory urography (HE 1.3 to 2.3 mSv.) at our departments. Only in obese patients with a body mass index of greater than 31 kg./m.2 is conventional unenhanced helical CT with higher radiation exposure recommended to achieve adequate image quality. PMID- 11912389 TI - Holmium: YAG laser endoureterotomy for ureterointestinal strictures. AB - PURPOSE: The management of ureterointestinal stricture in patients who have undergone urinary diversion can be challenging. Endourological techniques have been increasingly used in recent years for ureteral stricture. While long-term results may not be as reliable or durable as those of traditional open reconstructive surgical techniques, associated morbidity is much less. The holmium (Ho):YAG laser, which has cutting and coagulating properties, has been demonstrated to have many applications in urology. We report our experience with and long-term results of Ho:YAG laser endoureterotomy for ureterointestinal strictures. MATERIALS AND METHODS: We reviewed the charts and followup history of 23 patients in whom the Ho:YAG laser was used to treat ureterointestinal anastomotic stricture. Strictures were treated percutaneously via the antegrade approach with flexible endoscopes and the holmium laser. A reversed 12/6Fr endopyelotomy stent was left indwelling for 6 weeks postoperatively. Success was defined as symptomatic improvement and radiographic resolution of obstruction. RESULTS: Between 1993 and 2000, 23 patients with a mean age of 61 years underwent endo-ureterotomy using the Ho:YAG laser for 24 ureterointestinal stricture. An overall success rate of 71% (17 of 24 cases) was achieved at a mean followup of 22 months. The success rate of holmium laser endoureterotomy for ureterointestinal stricture at 1, 2 and 3 years was 85%, 72% and 56%, respectively. Seven patients had recurrent strictures of which 4 developed 16 months or more postoperatively. No complications were noted. CONCLUSIONS: Ho:YAG laser endoureterotomy for ureterointestinal stricture disease is a minimally invasive endourological procedure that may provide more durable results than other modalities used for endoureterotomy. The Ho:YAG laser with its ability to cut tissue precisely and provide hemostasis combined with its versatility and compatibility with flexible endoscopes is an ideal instrument for safely performing endoureterotomy. PMID- 11912390 TI - Cobalamin profiles in patients after urinary diversion. AB - PURPOSE: We assessed the risk of tissue cobalamin (vitamin B12) deficiency in patients after various types of urinary diversion. MATERIALS AND METHODS: Serum vitamin B12, methylmalonic acid and homocysteine were measured in 41 patients with urinary diversion, including an ileal neobladder in 12, ileal reservoir in 2, ileocecal reservoir in 10 and ileal conduit in 17. Followup was 0 to 10 years. RESULTS: The complete cobalamin profiles revealed a much higher incidence of tissue cobalamin deficiency in patients with each type of urinary diversion than measuring the vitamin B12 level alone. CONCLUSIONS: The risk of chronic tissue cobalamin deficiency after urinary diversion is higher than previously reported based on measuring serum vitamin B12 alone. Long-term patient monitoring for cobalamin deficiency or empirical supplemental therapy is indicated to prevent clinical cobalamin deficiency and irreversible sequelae. PMID- 11912391 TI - The patient with chronic epididymitis: characterization of an enigmatic syndrome. AB - PURPOSE: We provide a baseline description of men diagnosed with chronic epididymitis, explore relevant associations that may be important etiological factors and suggest a classification system and specific symptom assessment tool. MATERIALS AND METHODS: Men diagnosed with chronic epididymitis, described as symptoms of discomfort or pain at least 3 months in duration in the scrotum, testicle or epididymis localized to 1 or each epididymis on clinical examination, completed an extensive specific clinical inventory questionnaire. Evaluation included demographics, preceding and concurrent clinical history, duration since diagnosis, associated and previous clinical associations, frequency and severity of prostatitis, voiding and sexual symptoms, specific and general quality of life, and history of investigation and/or treatment for the condition. Volunteers with no past or concurrent history of chronic epididymitis completed similar clinical inventory questionnaires. RESULTS: A total of 50 consecutive men 21 to 83 years old (average age 46) diagnosed with chronic epididymitis who had an average symptom duration of 4.9 years (range 0.25 to 29) were enrolled in the study. The average pain score plus or minus standard deviation was 4.7 +/- 2.1 (range 0 to 10). Of the men 16% were reasonably satisfied with their quality of life. Although 66% of the patients thought about the symptoms some or a lot, in only 30% did symptoms keep them from doing the kinds of things that they would usually do. The most common previous therapies recollected by the patients were antibiotics (74%) and anti-inflammatory agents (36%). At the time of the survey 26% of the men were on some type of pain medication. There were no significant epidemiological, sexual, medical or associated factors that differentiated patients with chronic epididymitis from the 20 controls. A chronic epididymitis classification system (inflammatory, obstructive and epididymalgia) and a symptom assessment index based on assessing pain and quality of life-impact was developed. CONCLUSIONS: This comprehensive clinical survey of men diagnosed with chronic epididymitis is the first step for defining and characterizing this particular population. Development of a classification system and symptom assessment index may direct further studies in the etiology, epidemiology and management of chronic epididymitis. PMID- 11912392 TI - Does ultrastructural morphology of human detrusor smooth muscle cells characterize acute urinary retention? AB - PURPOSE: Acute urinary retention is relatively rare in otherwise healthy men. The condition may be attributable to a combination of obstruction and detrusor hypocontractility. We determined whether acute or chronic urinary retention is accompanied by characteristic ultrastructural features in bladder detrusor smooth muscle cells and whether any of these ultrastructural features may be useful for classifying these conditions. MATERIALS AND METHODS: Bladder biopsies of 15 men, including 13 in acute and 2 in chronic urinary retention, and 6 controls were examined by transmission electron microscopy. They were semiquantitatively and morphometrically analyzed for degenerative changes, cellular hypertrophy, variations in intercellular distances, abnormal cell junctions and configurations, and intracellular changes. RESULTS: No significant ultrastructural features were noted in the detrusor muscle of patients in acute urinary retention compared with controls. CONCLUSIONS: We cannot confirm the reports of others that characteristic ultrastructural features in smooth muscle cells accompany bladder outlet obstruction and detrusor hypocontractility. PMID- 11912394 TI - Anastomotic urethroplasty for bulbar urethral stricture: analysis of 168 patients. AB - PURPOSE: We reviewed our experience with anastomotic urethroplasty for anterior urethral stricture. MATERIALS AND METHODS: A chart review revealed 168 patients 6 to 82 years old (mean age 38) with at least 6 months of followup (mean 70, range 6 to 291) after anastomotic urethroplasty. RESULTS: Average stricture length was 1.7 cm. Of the 168 patients stricture recurred in 8 (5%) but was managed by direct vision internal urethrotomy or a single dilation in 5, while repeat urethroplasty was required in 3 (2%). In these 3 cases extenuating circumstances included patient dislodgment of the catheter with attempts to replace it that disrupted repair, a history of urethrocutaneous fistula and periurethral abscess, and previous irradiation complicating the stricture in 1 each. Other complications were uncommon, such as transient thigh pain or numbness in 3 patients (2%), small wound dehiscence in 2 (1%), and scrotal hematoma, erectile dysfunction and self-limited pulmonary edema in 1 (less than 1%) each. CONCLUSIONS: Anastomotic urethroplasty for anterior stricture has a high success rate of 95%. It is technically straightforward and complications are uncommon. Cure by anastomotic urethroplasty should be strongly favored over long-term management by direct vision internal urethrotomy or dilation. PMID- 11912393 TI - Intravesical capsaicin versus resiniferatoxin in patients with detrusor hyperreflexia: a prospective randomized study. AB - PURPOSE: Capsaicin and resiniferatoxin (Sigma Chemical Co., St. Louis, Missouri) administered intravesically are attractive options for treating detrusor hyperreflexia. Because the 2 agents differ in chemical structure and relative potency, possible differences in their clinical and urodynamic effects were investigated in this prospective comparative study. MATERIALS AND METHODS: A group of 24 spinal cord injured patients with refractory detrusor hyperreflexia were randomly assigned to receive a single dose of 2 mM. capsaicin in 30 ml. ethanol plus 70 ml. 0.9% sodium chloride or 100 nM. resiniferatoxin in 100 ml. 0.9% sodium chloride. Dwell time was 40 minutes with urodynamic monitoring. Urodynamics were performed at baseline before treatment, and after followups of 30 and 60 days. The frequency of daily catheterizations, incontinence episodes and side effects was recorded. RESULTS: There was no significant urodynamic or clinical improvement in the capsaicin arm at 30 and 60 days of followup. In the resiniferatoxin arm the mean uninhibited detrusor contraction threshold plus or minus standard deviation increased from 176 +/- 54 to 250 +/- 107 ml. at 30 days (p <0.05) and to 275 +/- 98 ml. at 60 days (p <0.01). Mean maximum bladder capacity increased from 196 +/- 75 to 365 +/- 113 ml. at 30 days (p <0.001) and to 357 +/- 101 ml. at 60 days (p <0.001). Daily catheterizations and incontinent episodes were significantly decreased at 30 and 60 days of followup. Autonomic dysreflexia, limb spasms, suprapubic discomfort and hematuria developed in most patients who received capsaicin but in none who received resiniferatoxin. CONCLUSIONS: Intravesical administration of resiniferatoxin is superior to that of capsaicin in terms of urodynamic results and clinical benefits in spinal cord injured patients and it does not cause the inflammatory side effects associated with capsaicin. PMID- 11912395 TI - Outcome of the artificial urinary sphincter in female patients. AB - PURPOSE: We reviewed the outcome in female patients at our unit in whom an artificial urinary sphincter was inserted. MATERIALS AND METHODS: We reviewed notes on 68 patients and mailed a questionnaire to those without recent followup. RESULTS: Median time since insertion was 12 years. Overall 25 patients (37%) had the original artificial urinary sphincter in situ and were dry at a median followup of 7 years. The artificial urinary sphincter was replaced for loss of function in 12 patients, of whom 11 were dry with the replaced device. The device was removed for erosion or infection in 31 patients, of whom 19 underwent successful replacement or were continent after removal. Overall 55 of 68 patients (81%) were continent. Those with neuropathic bladder dysfunction achieved a continence rate of greater than 90%, although half required sphincter removal initially. When the indication for insertion was stress incontinence, 70% of the patients had the original or a replaced artificial urinary sphincter in situ and 82% were continent. All patients with previous pelvic irradiation had the sphincter removed and urinary diversion was done. CONCLUSIONS: The overall continence rate in female patients after insertion of an artificial urinary sphincter is satisfactory. A satisfactory outcome was achieved in terms of stress incontinence and we would recommend an artificial urinary sphincter after an adequate anti-stress incontinence operation fails. Continence in patients with neuropathic bladder dysfunction is excellent and the artificial urinary sphincter should be considered first line treatment in this group, although the risk of revision surgery is high. Pelvic irradiation is a contraindication to the artificial urinary sphincter in female patients. PMID- 11912396 TI - Treatment of chronic prostatitis lowers serum prostate specific antigen. AB - PURPOSE: We evaluated men with documented chronic prostatitis and elevated serum prostate specific antigen (PSA) to determine whether treatment with antibiotics and anti-inflammatory drugs lowers serum PSA. MATERIALS AND METHODS: We retrospectively reviewed the records of 95 men who presented with serum PSA greater than 4 ng./ml. and were subsequently diagnosed with chronic prostatitis with greater than 10 white blood cells per high power field in expressed prostatic excretions. Patients meeting these criteria were treated with a 4-week course of antibiotics and a nonsteroidal anti-inflammatory agent. In all patients followup PSA was determined within 2 months of treatment. RESULTS: Mean PSA decreased 36.4% from 8.48 ng./ml. before to 5.39 after treatment (p <0.001). In 44 patients (46.3%) serum PSA decreased to below 4 ng./ml. (mean 2.48) and these patients no longer had an indication for biopsy. In the remaining 51 patients serum PSA remained elevated at greater than 4 ng./ml. and they underwent double sextant transrectal ultrasound guided biopsy. Pathological study showed prostate cancer in 13 cases (25.5%), chronic inflammation in 37 (72.5%) and only benign prostatic hypertrophy in 1 (1.05%). PSA in the 13 patients with prostate cancer decreased with treatment only 4.8% from 8.32 to 7.92 ng./ml. (p >0.05). Followup PSA at a mean of 11.4 months was determined in 19 of the 44 men who responded to treatment. Mean PSA increased only 4.5% from 2.35 to 2.46 ng./ml. (p >0.05) during this followup interval. CONCLUSIONS: In almost half of the patients diagnosed with elevated PSA and chronic prostatitis serum PSA normalized with treatment and there was no longer an indication for transrectal ultrasound guided biopsy. Our study suggests that chronic prostatitis is an important cause of elevated PSA and when it is identified, treatment can decrease the percent of negative biopsies. PMID- 11912397 TI - Baseline prostatic specific antigen does not predict the outcome of high energy transurethral microwave thermotherapy. AB - PURPOSE: We assessed the prognostic value of baseline prostate specific antigen (PSA) for outcome after high energy transurethral thermotherapy in patients with lower urinary tract symptoms. MATERIAL AND METHODS: Data were collected prospectively in 404 consecutive patients treated with high energy transurethral thermotherapy with the Prostatron device (EDAP-Technomed, Lyon, France). Patients were followed a minimum of 1 year. At baseline certain criteria were assessed, including pretreatment PSA, uroflowmetry, ultrasound measurement of prostatic volume, voided and post-void residual urine volume, and International Prostate Symptom Score (I-PSS) and quality of life scores. Outcome assessment included I PSS, quality of life score and uroflowmetry of peak urine flow. Linear regression analyses were performed to correlate baseline PSA with improved clinical parameters at 12 months of followup. Logistic regression analyses and receiver operating characteristics curves characterized the ability of baseline PSA to discriminate patients with a more or less favorable outcome. RESULTS: An evident linear association was identified for prostate size at baseline and PSA. After 1 year 36 patients were treated again due to transurethral thermotherapy failure and 16 had died, which was not related to lower urinary tract symptoms or treatment for lower urinary tract symptoms. To include re-treated patients in the analyses we considered that their I-PSS, quality of life and peak urine flow values at 1 year were unchanged compared with baseline. Of the 388 evaluable patients an improvement of 50% or more in I-PSS, quality of life and peak urine flow was observed in 57%, 62% and 44%, respectively. Absolute mean changes at 1 year were -9.7, -2 and 5.2 ml. per second for I-PSS, quality of life and peak urine flow, respectively. Neither linear nor logistic regression analysis showed any clinically relevant correlation between baseline PSA and changes in I-PSS (r = -0.004), quality of life (r = -0.135) or peak urine flow (r = 0.105) at 1 year. Receiver operating characteristics curves failed to distinguish more or less favorable outcomes in all evaluated parameters. CONCLUSIONS: Pretreatment PSA cannot predict the clinical outcome after high energy transurethral thermotherapy. PMID- 11912398 TI - Decreased suburethral prostatic microvessel density in finasteride treated prostates: a possible mechanism for reduced bleeding in benign prostatic hyperplasia. AB - PURPOSE: We evaluated the influence of finasteride on prostatic microvessel density to elucidate a mechanism of decreased bleeding in finasteride treated patients with hematuria secondary to benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 22 patients with clinical BPH and gross hematuria who underwent prostate reductive surgery between 1998 and 2000 were prospectively evaluated. The prostate from 10 finasteride treated and 12 untreated patients was immunohistochemically stained for CD-34. Microvessel density analysis was performed by quantifying positive stained blood vessels located within the stroma of hyperplastic nodules as well as in the suburethral portion of the prostate. RESULTS: Mean microvessel density plus or minus standard deviation in finasteride treated patients was significantly lower in the suburethral portion of the prostate versus untreated controls (14.0 +/- 2.8 versus 20.2 +/- 5.3 vessels per high power field, p <0.05). In the nodular hyperplasia there was no statistically significant difference in the treatment and control groups (mean 17.5 +/- 2.8 and 16.7 +/- 4.6 vessels per high power field, respectively). CONCLUSIONS: Finasteride significantly decreases suburethral prostatic microvessel density in patients with BPH, which may explain its efficacy for decreasing BPH associated bleeding. PMID- 11912399 TI - Long-term risk of re-treatment of patients using alpha-blockers for lower urinary tract symptoms. AB - PURPOSE: The efficacy of alpha-adrenoceptor blockers for the treatment of lower urinary tract symptoms has been proven in numerous studies. However, little is known about the efficacy of the longer term. We investigated the long-term risk of re-treatment in patients using alpha-adrenoceptor blockers for lower urinary tract symptoms and the parameters that influence this risk. MATERIALS AND METHODS: We reviewed the files of 316 patients with lower urinary tract symptoms treated at our department with the alpha-blockers terazosin, alfuzosin or tamsulosin. Using followup data up to 3 years, we calculated re-treatment percentages in each treatment group. Using extended followup of 5 years, we calculated the predictive value of various baseline characteristics for re treatment. RESULTS: The re-treatment rates were 27% for tamsulosin, 37% for alfuzosin and 49% for terazosin. The re-treatment rates of patients with mild, moderate and severe lower urinary tract symptoms were 27%, 33% and 70%, respectively. Patients with a maximum urine flow of less or more than 10 ml. per second had a re-treatment rate of 58% and 47%, respectively. Patients with a prostate volume of less or more than 40 ml. had a re-treatment rate of 48% and 72%, respectively. Patients who were urodynamically unobstructed versus obstructed patients had a re-treatment rate of 44% and 59%, respectively. CONCLUSIONS: Patients given alpha-blockers for lower urinary tract symptoms have a high risk of re-treatment. Tamsulosin has a markedly lower re-treatment percentage than alfuzosin and terazosin. Severe symptoms, poor urine flow, an enlarged prostate and urodynamically proven bladder outlet obstruction increase the risk of treatment failure. Preselection of the most suitable candidates for alpha-blockade may reduce this risk. PMID- 11912400 TI - Adult mullerian duct or utricle cyst: clinical significance and therapeutic management of 65 cases. AB - PURPOSE: We define guidelines for the exploration and treatment of adult mullerian duct cysts. MATERIALS AND METHODS: From January 1988 through September 1999 a diagnosis of enlarged prostatic utricle was made in 65 adults based on transrectal ultrasound findings. Echographic criteria to define simple versus complicated cysts were detailed. We reviewed the clinical presentation, diagnostic modalities, indications for invasive procedures and postoperative outcome. RESULTS: The usual clinical presentations were hematospermia in 40% of cases, other ejaculatory disturbances in 20%, recurrent testicular or pelviperineal pain in 33%, lower urinary tract irritation symptoms in 25%, lower urinary tract infection in 18.5%, male infertility in 12% and incidental finding in 18.5%. Cyst dimensions did not influence the indication for invasive procedures, which were performed in only 27 of the 65 patients (41.5%) to treat disabling symptoms in 28% and obstructive infertility in 5%, and investigate complicated cysts on transrectal ultrasound in 6%. These procedures included transperineal or transrectal puncture in 9 patients, simple endoscopic section of the utricle meatus in 12 and large marsupialisation in 6. Complete and sustained cure was noted in half of the patients treated with cyst puncture only, although echographic relapse was the rule. Endoscopic procedures definitely improved or cured 82% of the patients at a mean followup of 51 months, during which neither early nor late complications were noted. CONCLUSIONS: Since almost 60% of adults diagnosed with a mullerian duct cyst did not experience any cyst related symptoms or ejaculatory-fertility impairment, we recommend that investigation and/or treatment should only be done in symptomatic or infertile patients. PMID- 11912401 TI - The relationship of serum testosterone to erectile function in normal aging men. AB - PURPOSE: We evaluated the variation in serum testosterone in normal aging men and its relationship with erectile function. MATERIALS AND METHODS: In a study that was not community based and during a free screening program for prostate cancer 1,071 men were invited to complete a sexual activity questionnaire, that is the abridged 5-item version of the International Index of Erectile Function (IIEF-5), as a diagnostic tool for erectile dysfunction. Possible scores on the IIEF-5 are 1 to 25 and erectile dysfunction was classified into 5 categories based on the scores, namely severe-1 to 7, moderate-8 to 11, mild to moderate-12 to 16, mild 17 to 21 and none-22 to 25. Serum total testosterone was measured between 8:00 and 10:00 a.m. in all men. RESULTS: Of the 1,071 men 965 (90.1%) were included in this study, of whom 88% were white and 12% were black. Mean age was 60.7 years. In this sample the prevalence of all degrees of erectile dysfunction was estimated to be 53.9%. The degree of erectile dysfunction was mild in 21.5% of cases, mild to moderate in 14.1%, moderate in 6.3% and severe in 11.9%. According to age the erectile dysfunction rate was 36.4% in the 40 to 49, 42.5% in the 50 to 59, 58.1% in the 60 to 69, 79.4% in the 70 to 79 and 100% in the 80 years and older groups (p <0.05). The variation in mean serum total testosterone in the age groups was not statistically significantly different (p >0.05). Pearson coefficients of age and total testosterone did not reveal any significant correlation (r = 0.00376, p = 0.907), similar to IIEF-5 score and total testosterone (r = 0.0163, p = 0.612). However, analysis of the variables IIEF-5 and age showed a statistically significant inverse or negative relationship (r = 0.3449, p <0.05). CONCLUSIONS: Erectile dysfunction showed a clear association with aging but no consistent correlation of total testosterone with erectile condition was identified. PMID- 11912402 TI - Can varicocelectomy significantly change the way couples use assisted reproductive technologies? AB - PURPOSE: We assessed how varicocelectomy alters semen quality in a large cohort of infertile men and determined whether it can change patient candidacy for assisted reproductive technology procedures. MATERIALS AND METHODS: A cohort of 540 infertile men with clinical palpable varicocele underwent microsurgical varicocelectomy and were followed more than 1 and 2 years postoperatively for alterations in semen quality and conception, respectively. Preoperatively and postoperatively the total motile sperm count was calculated in all semen analyses. Based on total motile sperm count values patients were divided into 4 groups according to the type of assisted reproductive technology for which they qualified, including 0 to 1.5 million or less (intracytoplasmic sperm injection candidates), 1.5 to 5 million or less (in vitro fertilization candidates), 5 to less than 20 million (intrauterine insemination candidates) and 20 million or greater sperm (spontaneous pregnancy candidates). Preoperative and postoperative semen quality was compared among individuals in these cohorts to determine the shifts in assisted reproductive technology care that are possible after varicolectomy. RESULTS: Mean patient age was 29.5 years (range 18 to 58). Microsurgical varicocelectomy was bilateral in 393 patients (73%), on the left side in 146 (27%) and on the right side in 1 (0.2%). A positive response to varicocelectomy, defined as a greater than 50% increase in total motile sperm count, was observed in 271 patients (50%). An overall spontaneous pregnancy rate of 36.6% was achieved after varicocelectomy with a mean time to conception of 7 months (range 1 to 19). Of preoperative in vitro fertilization and intracytoplasmic sperm injection candidates 31% became intrauterine insemination or spontaneous pregnancy candidates after varicolectomy. Of intrauterine insemination candidates 42% gained the potential for spontaneous pregnancy. CONCLUSIONS: Varicocelectomy has significant potential not only to obviate the need for assisted reproductive technology, but also to down stage or shift the level of assisted reproductive technology needed to bypass male factor infertility. PMID- 11912403 TI - Outcome of in vitro fertilization and intracytoplasmic injection of epididymal and testicular sperm obtained from patients with obstructive and nonobstructive azoospermia. AB - PURPOSE: We assessed fertilization, pregnancy and miscarriage rates in patients with obstructive and nonobstructive azoospermia who underwent intracytoplasmic sperm injection. MATERIALS AND METHODS: From June 1996 to March 2000, 166 consecutive patients (198 intracytoplasmic sperm injection cycles) with azoospermia were studied. Of these 198 cycles 68 were performed due to nonobstructive azoospermia using testicular spermatozoa and 130 were performed due to obstructive azoospermia using epididymal spermatozoa. RESULTS: The normal (2 pronuclei) and abnormal (1 plus 3 pronuclei) fertilization rates for obstructive and nonobstructive azoospermia were 60.5% and 16.6%, and 54% and 16.4%, respectively (p >0.05). The pregnancy rate per cycle, pregnancy rate per patient and abortion rate were 30%, 39.8% and 28% for obstructive azoospermia, and 22%, 28.3% and 40% for nonobstructive azoospermia (p <0.05). The normal and abnormal fertilization rates were 58.7% and 21.4% for percutaneous epididymal sperm aspiration (PESA), 62.3% and 10.4% for PESA plus testicular sperm aspiration (TESA), and 57.3% and 14.5% for TESA, respectively (p >0.05). The pregnancy rate per cycle, pregnancy rate per patient and abortion rate were 34.6%, 54.5% and 11.1% for PESA, 37.5%, 37.5% and 33.3% for PESA plus TESA, and 26.1%, 31% and 41% for TESA, respectively (PESA versus PESA plus TESA p >0.05, and PESA and PESA plus TESA versus TESA p <0.05). Epididymal or testicular motile sperm resulted in a lower abortion rate than epididymal or testicular immotile sperm (p = 0.03). CONCLUSIONS: No differences were noted in the fertilization and embryo transfer rates irrespective of etiology (obstructive versus nonobstructive) and type of spermatozoa (epididymal versus testicular). Testicular sperm retrieval results in lower fertilization and pregnancy rates as well as higher abortion rates than epididymal sperm retrieval. PMID- 11912404 TI - Financial analysis of open versus laparoscopic radical nephrectomy and nephroureterectomy. AB - PURPOSE: Laparoscopic radical nephrectomy and nephroureterectomy are rapidly becoming established procedures in select patients with renal cell carcinoma and upper tract transitional cell carcinoma, respectively. We present a retrospective comparative analysis of laparoscopic versus open radical nephrectomy and nephroureterectomy from a financial standpoint. The effect of the learning curve on costs incurred was also evaluated. MATERIALS AND METHODS: Detailed itemized cost data on 18 contemporary cases of open radical nephrectomy performed from September 1997 to July 1998 were compared with similar data on 20 initial laparoscopic cases performed from September 1997 to July 1998 and 15 more recent laparoscopic radical nephrectomy cases performed from August 1998 to July 1999. Financial data were also compared on 14 contemporary patients each who underwent open radical nephroureterectomy from June 1997 to December 1999, initial laparoscopic radical nephroureterectomy from June 1997 to December 1998 and more recent laparoscopic radical nephroureterectomy from January 1999 to October 2000. Yearly financial costs were adjusted for inflation by a 4% annual rate to reflect year 2000 data. RESULTS: For radical nephrectomy mean operative time in the 18 open, 20 initial laparoscopic and 15 recent laparoscopic cases was 185.3, 205.7 and 147.3 minutes, respectively. Mean specimen weight was 555, 616 and 558 gm., and mean hospital stay was 132, 31 and 23 hours, respectively. Compared with open radical nephrectomy mean total costs associated with initial laparoscopy were 33% greater (p = 0.0003). Mean intraoperative costs were 102% greater and mean postoperative costs were 50% less. In contrast, the more recent laparoscopic cases were an overall mean of 12% less expensive than open surgery (p = 0.05). Mean intraoperative costs were only 33% greater and mean postoperative costs were 68% less. For radical nephroureterectomy mean operative time in the 14 open, 14 initial laparoscopic and 14 recent laparoscopic cases was 246, 196 and 195 minutes, respectively. Mean specimen weight was 442, 517 and 531 gm., and mean hospital stay was 142, 63 and 32 hours, respectively. Compared with open radical nephroureterectomy mean total costs associated with initial laparoscopic cases were 28% greater (p = 0.03). Mean intraoperative costs were 65% greater and mean postoperative costs were 27% less. In contrast, the more recent laparoscopic cases were an overall mean of 6% less expensive than open surgery (p = 0.63). Mean intraoperative costs were only 31% greater and mean postoperative costs were 62% less. CONCLUSIONS: Initially in the learning curve laparoscopic radical nephrectomy and nephroureterectomy were 33% and 28% financially more expensive, respectively, than their open counterparts. However, with increased operator experience and efficiency resulting in more rapid operative time and decreased hospitalization laparoscopic radical nephrectomy and nephroureterectomy are currently 12% and 6% less expensive, respectively, than their open counterparts at our institution. PMID- 11912405 TI - Depressive symptoms and quality of life in patients with interstitial cystitis. AB - PURPOSE: Previous research suggests that patients with interstitial cystitis have poorer quality of life and higher levels of depressive symptoms. However, most studies to date have been limited by the lack of standard measures to describe the experience of living with interstitial cystitis. In addition, to our knowledge no study has used a structured interview to assess depressive symptomatology. We investigated the extent of depressive symptoms and impaired quality of life in a sample of female patients with interstitial cystitis compared with healthy controls. Relationships among physician rated symptom severity, quality of life and depressive symptoms were also examined. MATERIALS AND METHODS: At a clinic visit 65 female patients previously diagnosed with interstitial cystitis and 40 age matched, healthy controls completed questionnaires on depressive symptoms (Beck Depression Inventory) and quality of life (Medical Outcomes Study 36-Item Short Form), and a structured interview on depressive symptoms (Hamilton Rating Scale for Depression) with trained interviewers. RESULTS: Patients reported compromised quality of life compared with healthy controls across various domains, including physical functioning, ability to function in one's normal role and vitality. They also had more severe depressive symptoms on the 2 depression measures. In patients, greater interstitial cystitis severity was associated with greater compromise in physical and social functioning, and mental health but not in other quality of life domains or depressive symptoms. CONCLUSIONS: A diagnosis of interstitial cystitis is related to poorer functioning in various life domains. Decrements increase with disease severity. PMID- 11912406 TI - Primary care and urology patients with the male pelvic pain syndrome: symptoms and quality of life. AB - PURPOSE: We assessed symptoms and health related quality of life in men who received prostatitis-prostatodynia diagnoses at primary care and urology visits, and compared those in whom pain-discomfort had versus had not resolved approximately 1 month later. MATERIALS AND METHODS: Telephone interviews were done with 357 men an average of 1 month after a prostatitis-prostatodynia diagnosis was made at a health maintenance organization visit. The interview included the National Institutes of Health Chronic Prostatitis Symptom Index, and pain and health related quality of life measures. RESULTS: The most common pain location was the pubic-bladder area. Mean scores on most health related quality of life measures were below average, and higher pelvic pain and urinary symptom scores were associated with worse quality of life. This episode of pelvic pain was the first lifetime episode in fewer urology (22%) than primary care (38%) patients (p = 0.02). Urology patients had longer symptom episodes (p = 0.000), more days with pain in the last month (p = 0.002) and higher National Institutes of Health Chronic Prostatitis Symptom Index pain scores (p = 0.002). Men with pain in the testicles, penis or between the rectum and testicles at the visit, and with longer symptom duration before the visit were significantly more likely to have continued pain between the visit and interview. CONCLUSIONS: Pelvic pain is often a persistent, recurrent condition that can have a significant negative impact on quality of life. The average symptom severity in men with pelvic pain in primary care and urology settings is lower than that in tertiary care samples. PMID- 11912407 TI - Fibrin sealant for the reconstruction of fournier's gangrene sequelae. AB - PURPOSE: We describe the use of fibrin tissue adhesive as an adjunct for reconstructing genital skin loss due to Fournier's gangrene. MATERIALS AND METHODS: We treated 2 patients with Fournier's gangrene with repeat surgical debridement and antibiotics. Delayed primary closure was enhanced by using liquid fibrin sealant. In 1 case the sealant was used to obliterate a large testicular thigh pouch that had become infected. In the other case it was used to anchor the under surface of a thigh flap for scrotal reconstruction. RESULTS: In each patient the fibrin tissue adhesive prevented further complications of Fournier's disease. CONCLUSIONS: Fibrin sealant is an effective adjunct for managing extensive genital skin loss caused by Fournier's gangrene. PMID- 11912408 TI - Giant lipoma of the adrenal gland. PMID- 11912409 TI - Delayed transcaval renal penetration of a Greenfield filter presenting as symptomatic hydronephrosis. PMID- 11912410 TI - Adenocarcinoma in a colon conduit. PMID- 11912411 TI - Primary choriocarcinoma of the bladder with the detection of isochromosome 12p. PMID- 11912412 TI - Adenocarcinoma arising from a defunctionalized bladder. PMID- 11912413 TI - Recurrent giant cell carcinoma of the bladder. PMID- 11912414 TI - Angiosarcoma of the penis masquerading as a Peyronie's plaque. PMID- 11912415 TI - Spontaneous rupture of a testicular tumor. PMID- 11912416 TI - Primary choriocarcinoma in the posterior mediastinum. PMID- 11912417 TI - Bilateral testicular carcinoma in patient with the persistent mullerian duct syndrome. PMID- 11912418 TI - Primitive neuroectodermal tumor of the spermatic cord. PMID- 11912419 TI - Does PC-SPEs interact with warfarin? PMID- 11912420 TI - An easy method to localize the vesical opening of an enterovesical fistula. PMID- 11912421 TI - Eosinophilic cystitis presenting as a recurrent symptomatic bladder mass following intravesical mitomycin C therapy. PMID- 11912422 TI - Giant urethral diverticulum: a novel approach to repair. PMID- 11912423 TI - Does the outpatient setting provide the best environment for medical student learning of urology? AB - PURPOSE: To determine in which environment medical students learn clinical urology most effectively, we retrospectively reviewed a natural experiment in which medical students were randomly assigned to complete the 1-week rotation in clinical urology in an outpatient/clinic based or inpatient/operative setting. MATERIALS AND METHODS: Exit surveys were completed by 25 of the 39 medical students (64%) who had just completed the mandatory 1-week rotation in urology. Students were asked to record on a 5-point scale the amount learned in regard to 13 urological topics and skills, and to document the number of patient encounters experienced per topic and skill. RESULTS: Students randomized to the outpatient/clinic based setting tended to be 1) more likely to have exposure to a greater number and breadth of patients with common urological problems, 2) more likely to perform pertinent physical examination skills and 3) more likely to perceive that they learned more about a given curricular topic or skill. CONCLUSIONS: Our data indicate that the outpatient/clinic based setting may be a higher yield environment for medical students learning clinical urology compared with the inpatient/operative setting. The development of a validated means to assess actual student learning in clinical urology is needed to confirm these findings. PMID- 11912424 TI - Re: Gastric patch pyeloplasty: development of an animal model to produce upper tract urinary acidification for treating struvite urinary calculi. PMID- 11912425 TI - Re: Transperitoneal preperitoneal laparoscopic lumbar incisional herniorrhaphy. PMID- 11912426 TI - Re: Inguinal hernia after radical retropubic prostatectomy for prostate cancer: a study of incidence and risk factors in comparison to no operation and lymphadenectomy. PMID- 11912428 TI - Re: Complications of ureteroscopy: analysis of predictive factors. PMID- 11912427 TI - Re: Prostate cancer: a brief history and the discovery of hormonal ablation treatment. PMID- 11912429 TI - Re: optimal prevention and management of proximal ureteral stent migration and remigration. PMID- 11912430 TI - Re: Penile injury. PMID- 11912431 TI - Re: sexuality preserving cystectomy and neobladder: initial results. PMID- 11912432 TI - Re: a prospective randomized clinical trial to evaluate methods of postoperative care of hypospadias. PMID- 11912433 TI - Re: The natural history of idiopathic urethrorrhagia in boys. PMID- 11912434 TI - Re: Particle migration after transurethral injection of carbon coated beads for stress urinary incontinence. PMID- 11912435 TI - Treatment of vesicoureteric reflux by endoscopic injection of Teflon. 1984. AB - Thirteen girls with grade III-V vesicoureteric reflux were treated by endoscopic injection of Teflon paste behind the intravesical ureter. Fourteen of the 18 treated ureters showed complete absence of reflux after one injection of Teflon. Three ureters required a second injection of Teflon for successful treatment of the reflux. One ureter with grade IV reflux was converted to grade II reflux. Properly carried out, this procedure corrects reflux. It takes less than 15 minutes, may be done as a day procedure, and avoids open surgery. There have been no complications. (Reprinted with permission from Br Med J, 289: 7-9, 1984) PMID- 11912437 TI - Simple surgical repair of bilateral renal artery stenosis in a patient with neurofibromatosis. PMID- 11912438 TI - Ureteroscopy for the treatment of urolithiasis in children. AB - PURPOSE: Ureteroscopy for treating urolithiasis in prepubertal children has become more common with the advent of smaller instruments. We reviewed our experience with ureteroscopy for urolithiasis in this cohort of patients as well as the literature using this treatment modality in children. MATERIALS AND METHODS: Between 1994 and 2000 we performed 27 ureteroscopic stone extractions in 25 children. Ureteroscopy was done in a manner similar to that in adults. Ureteral dilation was performed when necessary to access the ureter. A stent was placed postoperatively if there was significant ureteral trauma. RESULTS: Of the 25 children 13 were male and 12 were female. Average age was 9.2 years (range 3 to 14). Stones were 2 to 12 mm. in greatest diameter (average 6). Of the 27 procedures the ureteral orifice was dilated before stone treatment in 15 (56%), while in 19 (70%) a stent was placed afterward. No intraoperative and 2 postoperative complications were identified. Overall 92% of the children were rendered stone-free after 1 procedure and 100% were stone-free after 2. CONCLUSIONS: Ureteroscopy for urolithiasis in prepubertal children is safe and effective. Routine ureteral dilation and ureteral stent placement are not always necessary in these patients. PMID- 11912439 TI - Laparoscopically assisted correction of transverse testicular ectopia. PMID- 11912440 TI - Laparoscopic variability of the internal inguinal ring: review of anatomical variation in children with and without a patent processus vaginalis. AB - PURPOSE: Diagnostic laparoscopy is routinely performed at our institution in children with a unilateral inguinal hernia to determine whether the contralateral processus vaginalis is patent. We reviewed the anatomical variability of the inguinal ring at laparoscopy in children with and without a hernia in various age groups. MATERIALS AND METHODS: Since 1992, we have performed diagnostic laparoscopy in more than 1,500 children with a known inguinal hernia. Intraoperative imaging was correlated with clinical and operative findings to characterize the anatomical variability of the internal ring. RESULTS: The internal ring had many variations. The photographs presented show evidence of the progression from the flat closed ring to the widely open sac. Clefts and veils of peritoneum sometimes made determining the exact anatomy difficult. Experience shows that the various anatomical variations are associated with different pathological conditions. CONCLUSIONS: Characterization of the anatomical variability of the internal ring is essential for determining the patency of the processus vaginalis at diagnostic laparoscopy in children with a known inguinal hernia. We classified these variations in accordance with clinical and surgical findings. PMID- 11912441 TI - Minimally invasive extravesical ureteral reimplantation for vesicoureteral reflux. AB - PURPOSE: We designed a new extravesical ureteral reimplantation technique with a minimally invasive approach from skin to ureterovesical junction with less perivesical tissue manipulation to avoid extensive bladder denervation. MATERIALS AND METHODS: Between July 1996 and December 2000, 37 boys and 52 girls 1.2 to 10.8 years old (mean age plus or minus standard deviation 3.8 +/- 2.5) (113 ureters) were treated with minimally invasive extravesical ureteral reimplantation. Vesicoureteral reflux was graded I to V in 8, 12, 43, 29 and 21 cases, respectively. The technique involves an approximately 10 to 15 mm. incision passing through the small triangular gap of the aponeurosis of the external abdominal oblique muscle and transversalis fascia to the point of the ureterovesical junction. The surgical field was exposed with mini-retractors and fine dissecting instruments were used to avoid unnecessary tissue manipulation. RESULTS: At postoperative followup 1 patient had persistent grade II reflux and 2 had moderate hydronephrosis and hydroureter, which resolved after 18 months. No patient returned due to voiding inefficiency or for pain control after discharge from the outpatient setting. CONCLUSIONS: This new technique can be easily used for vesicoureteral reflux with the advantages of simple intervention for surgeons, especially those with inguinal herniorrhaphy and antireflux surgery experience, and less wound discomfort for patients. The whole procedure can be performed on an outpatient basis. However, the decision to use this technique should be based on individual consideration. PMID- 11912442 TI - Leydig cell function after cryptorchidism: evidence of the beneficial result of early surgery. AB - PURPOSE: We determined whether there are differences indicative of the function of the Leydig cell-pituitary axis in formerly unilateral cryptorchid men. Parameters were compared with those in control men. MATERIALS AND METHODS: We determined luteinizing hormone, testosterone, free testosterone, follicle stimulating hormone, inhibin B, sperm density, motility and morphology, testicular volume, patient weight and age at orchiopexy or at other nonrelated childhood surgery in controls. RESULTS: Significant correlations were noted between hormone levels and other parameters. An inverse correlation between age of surgery and testosterone suggested the detrimental effect of deferring orchiopexy during childhood. Correlations of testosterone with sperm density, motility and morphology suggested a direct relationship between spermatogenesis and testosterone in cryptorchid men. An inverse relationship between body weight and testosterone was observed in each group. There was considerable variation in the range of parameters, sometimes extending into the abnormal range. However, results indicated no differences in mean free testosterone, testosterone or luteinizing hormone in the formerly cryptorchid and control groups, and no difference in time to conception in fertile, formerly cryptorchid men and controls. Furthermore, there were no differences in hormone levels in fertile (as indicated by paternity) and infertile formerly cryptorchid men. CONCLUSIONS: This study suggests that men who underwent orchiopexy in later childhood have subclinically decreased Leydig cell function. It may result in a less than optimal hormonal milieu for adult reproductive function. These data provide support for orchiopexy during infancy to preserve Leydig cell function and, thereby, potentially enhance fertility. PMID- 11912443 TI - SRY gene expression in the ovotestes of XX true hermaphrodites. AB - PURPOSE: The pathogenesis of 46 XX true hermaphroditism is uncertain and the role of the SRY gene in ovotestis development has not been thoroughly evaluated. We ascertained the presence of the SRY gene and SRY protein in the ovotestis. MATERIALS AND METHODS: We evaluated 8 ovotestes by cytogenetic analysis of fibroblast cell culture and analysis of gonadal tissue by polymerase chain reaction to detect the SRY gene and by immunohistochemistry with a monoclonal antibody to human recombinant SRY protein. RESULTS: Fibroblast culture of the ovotestes demonstrated a 46XX karyotype. By polymerase chain reaction all 8 ovotestes demonstrated the SRY gene at low levels. By immunohistochemistry SRY protein was detected in all ovotestes, predominantly in Sertoli and germ cells. CONCLUSIONS: The SRY gene has a role in ovotestis genesis. Mosaicism with a Y bearing cell line in the gonad is a possible explanation and further study is warranted. PMID- 11912445 TI - Urinary crystallization inhibitors do not prevent crystal binding. AB - PURPOSE: Renal stone formation requires the persistent retention of crystals in the kidney. Calcium oxalate monohydrate (COM) crystal binding to Madin Darby canine kidney strain I (MDCK-I), a cell line that resembles the epithelium in the renal distal tubule/collecting duct, is developmentally regulated, while LLC-PK1 cells (American Type Tissue Collection), which are widely used as a model of the renal proximal tubule, bind crystals irrespective of their stage of epithelial development. Whereas to our knowledge the binding molecules for COM at the surface of LLC-PK1 cells are still unknown, crystals adhere to the hyaluronan (HA) rich pericellular matrix transiently expressed by mobile MDCK-I cells. In the current study we investigated whether crystal binding to either cell type is influenced by urinary substances, including glycoprotein inhibitors of crystallization MATERIALS AND METHODS: We studied crystal binding to MDCK-I cells during wound repair, to confluent LLC-PK1 cells and to HA immobilized on a solid surface using [14C] COM pretreated or not pretreated with urine from healthy male volunteers. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis were performed to assess whether the crystals became coated with urine derived proteins RESULTS: Western blot analysis demonstrated that pretreated COM crystals were covered with protein inhibitors of crystallization. However, this protein coat had no significant effect on the level of crystal binding to either cell type. In contrast, the adherence of urine treated crystals to immobilized HA was significantly reduced CONCLUSIONS: The adherence of crystals to pericellular matrixes may encompass more than their simple fixation to the polysaccharide HA. Calcium oxalate crystal retention is not prevented by coating crystals with urinary constituents such as glycoproteins and, therefore, may predominantly depend on the surface properties of the renal tubular epithelium. PMID- 11912444 TI - Cell adhesion proteins as tumor suppressors. AB - PURPOSE: We summarize recent progress on the role of cell adhesion molecules in biology and discuss the potential application of cell adhesion molecules for managing urological cancer. MATERIALS AND METHODS: We comprehensively reviewed the literature from 1982 to 2001, including peer reviewed publications and recent abstracts from national meetings, relevant to cell adhesion molecules in urological cancer. RESULTS: A growing body of evidence suggests that alterations in the adhesion properties of neoplastic cells have a pivotal role in the development and progression of cancer. Loss of intercellular adhesion and desquamation of cells from the underlying lamina propria allows malignant cells to escape from their site of origin, degrade the extracellular matrix, acquire a more motile and invasion phenotype, and invade and metastasize. In addition to participating in tumor invasiveness and metastasis, adhesion molecules regulate or significantly contribute to various functions, including signal transduction, cell growth, differentiation, site specific gene expression, morphogenesis, immunological function, cell motility, wound healing and inflammation. To date a diverse system of transmembrane glycoproteins has been identified that mediates cell-cell and cell-extracellular matrix adhesion. The main families of adhesion molecules include members of the Ig superfamily, cadherins, integrins and selectins. CONCLUSIONS: Multiple and diverse cell adhesion molecules participate in intercellular and cell-extracellular matrix interactions of cancer. Cancer progression is a multistep process, in which some adhesion molecules have a pivotal role in the development of recurrent, invasive and distant metastasis. Recent data implicate some of these molecules in cell signaling and tumor suppression, which has important consequences for tumor growth. PMID- 11912446 TI - The fluorescence biodistribution and kinetics of aminolevulinic acid induced protoporphyrin IX in the bladder of a rat model with orthotopic urothelial carcinoma. AB - PURPOSE: Photodynamic therapy is an alternative intravesical therapy modality for superficial bladder cancer. Aminolevulinic acid (ALA) induces the production of the endogenous photosensitizer protoporphyrin IX (PpIX). We compared intravenous versus intravesical administration of ALA and established the proper timing and dose of ALA for photodynamic therapy. To characterize the distribution of ALA in rat bladder tumor and normal bladder layers a cooled charge coupled device camera was used. MATERIALS AND METHODS: A total of 40 female Fisher F344 rats were used as test animals, including 36 inoculated with AY-27 tumor cells intravesically. PpIX accumulation was investigated by fluorescence microscopy comparing 100 and 300 mg./kg. intravenous administration with a 100 mg./ml. intravesical dose of ALA. Three areas of urothelium, submucosa and muscularis of the bladder wall were chosen for analysis. The software used allowed semiquantitative analysis by calculating the mean fluorescence count plus or minus standard error of mean within any chosen area on the fluorescence image. RESULTS: In this study the highest fluorescence difference in PpIX accumulation in tumor and the normal epithelium to the muscularis layer was achieved at 2 hours with intravenous administration (7:1 to 50:1). The highest absolute fluorescence levels were observed at 2 hours with the 100 mg./kg. intravenous dose and at 4 hours with the higher 300 mg./kg. dose. The difference in fluorescence intensity in tumor tissue to normal urothelium was 2:1 to 3:1 at 2 hours. At 4 hours it was less than 2:1. After intravesical administration no difference in PpIX accumulation in tumor and normal urothelium was observed. However, there was a 7:1 ratio in regard to the muscularis layer at 4 hours. CONCLUSIONS: According to the results of this study a difference in PpIX accumulation in urothelial carcinoma or normal urothelium and the muscular layer of the bladder can be achieved by each route of ALA administration. Although intravesical installation provided tumor and normal urothelium labeling comparable to the intravenous route, it lost the selectivity of PpIX accumulation in tumor and normal urothelium. The effect of this finding on clinical therapy results remains to be resolved in the future. PMID- 11912447 TI - The use of real-time quantitative polymerase chain reaction to detect hypermethylation of the CpG islands in the promoter region flanking the GSTP1 gene to diagnose prostate carcinoma. AB - PURPOSE: We developed a real-time, quantitative, methylation sensitive polymerase chain reaction (PCR) protocol to analyze hypermethylation of the CpG islands in the promoter region of the pi class glutathione-S-transferase gene GSTP1 in prostate cancer tissue. MATERIALS AND METHODS: A total of 21 prostate cancer and 72 benign prostate hyperplasia (BPH) tissue samples were analyzed. Genomic DNA was digested with restriction enzyme, followed by real-time quantitative PCR amplification. Cycle threshold values were used to determine whether cancer genome was present in these tissues. A cutoff cycle threshold value of 35 was arbitrarily assigned. Samples with a cycle threshold of 35 or less were considered positive for prostate cancer. Conventional nested PCR was also performed for comparison. RESULTS: The mean cycle threshold values plus or minus standard deviation in prostate cancer and BPH cases were 30.12 +/- 2.88 and 37.77 +/- 2.72, respectively. All prostate cancer samples analyzed showed positive results, while 5 of the 72 BPH samples tested positive. Conventional nested PCR data indicated that 19 of 21 prostate cancer cases were positive for the methylation change, while 71 of 72 BPH cases tested negative. The test limitations of real-time PCR and the nested PCR protocols were determined to be 0.048 and 0.64 ng. DNA, respectively. CONCLUSIONS: We established a novel protocol for detecting the methylation change in the 5' regulatory sequence flanking the GSTP1 gene. The sensitivity of this protocol was superior to that of conventional nested PCR. The data also suggest that this novel protocol may accurately discriminate prostate carcinoma from BPH. PMID- 11912448 TI - Selective activation of members of the signal transducers and activators of transcription family in prostate carcinoma. AB - PURPOSE: Cytokines, hormones and growth factors use signal transducers and activators of transcription (STAT) signaling pathways to control various biological responses, including development, differentiation, cell proliferation and survival. Constitutive activation of STATs has been found in a wide variety of human tumors. In this study we examined the activity of STATs in primary human prostate tissues. MATERIALS AND METHODS: STAT activity was determined in 104 human primary prostate tissues, including 42 tumors, 42 matched normal prostates adjacent to tumors and 20 normal prostates from donors without cancer by electromobility shift assay. RESULTS: Significant levels of activated Stat4 and Stat6 were detected in primary prostate tissues. However, little or no expression of active Stat1, Stat2 or Stat5 was detected in primary prostate tissues. Significantly higher levels of constitutive Stat6 activity were found in prostate carcinomas compared with levels in normal tissue adjacent to tumors and normal prostates from donors without prostate cancer. There was no significant difference in Stat6 activity in normal prostate tissues adjacent to tumors and normal prostates from donors without prostate cancer. The levels of Stat4 activity varied but failed to yield statistically significant differences among tumors, matched normal prostates adjacent to tumors and normal prostates from donors without cancer. CONCLUSIONS: We have previously shown that Stat3 is activated in prostate cancer. The results of the current study demonstrate that in addition to Stat3, Stat6 is selectively activated in prostate cancer. PMID- 11912449 TI - Metabolic gaps in bone formation may be a novel marker to monitor the osseous metastasis of prostate cancer. AB - PURPOSE: Although skeletal metastasis is a common complication associated with prostate cancer, assessment of the response of osseous metastasis to therapy is still difficult. To remedy this situation a study of bone formation metabolic markers gaps in patients with prostate cancer and osseous metastasis was done to determine these gaps as clinical markers for the efficacy of therapy against osseous metastasis. MATERIALS AND METHODS: The activities of the bone formation markers amino terminal propeptide of type I procollagen (PINP), bone alkaline phosphatase (BAP) and osteocalcin (OC) were measured in 57 and 60 patients with prostate cancer without and with osseous metastasis, respectively. Of the 60 patients with osseous metastasis serial measurements were performed in 31. The status of osseous metastasis at the time of marker measurement was categorized, and the BAP-to-PINP, OC-to-PINP and OC-to-BAP ratios were calculated. RESULTS: The BAP-to-PINP ratio did not change. However, the OC-to-PINP and OC-to-BAP ratios were significantly different depending on osseous metastasis status. The ratios were high in patients with no osseous metastasis or with metastasis shrinkage (improvement) but low in those in whom metastases were growing and/or spreading (progression). Cutoff levels of OC-to-PINP and OC-to-BAP ratios were determined by comparing ratios with clinical judgment on the response to treatment in all patients. When these cutoffs were applied to the 31 patients, the OC-to-PINP ratio agreed with clinical judgment in 29 (94%) and the OC-to-BAP ratio agreed in 28 (90%). CONCLUSIONS: Different bone formation markers seem to be sensitive clinical markers for judging the response to therapy of prostate cancer metastasized to bone. PMID- 11912450 TI - In vitro biocompatibility evaluation of naturally derived and synthetic biomaterials using normal human bladder smooth muscle cells. AB - PURPOSE: Tissue engineering of the urinary tract often requires the use of various biomaterials. Adequate biomaterial biocompatibility is necessary for successful urinary reconstruction. In this study using a primary normal human bladder smooth muscle cell culture system we evaluated the in vitro biocompatibility of a number of naturally derived biomaterials, including bladder submucosa, small intestinal submucosa, collagen and alginate, and polymeric biomaterials, including polyglycolic acid, poly(L-lactic acid) and poly(lactic-co glycolic acid, which have been used for urinary reconstruction experimentally or clinically. MATERIALS AND METHODS: To determine the cytotoxic and bioactive effects of bladder submucosa, small intestinal submucosa, collagen, alginate, polyglycolic acid, poly(L-lactic acid) and poly(lactic-co-glycolic acid) we measured cell viability, metabolic activity, apoptotic properties and DNA synthesis activity with 4 types of assays, namely Neutral Red (Sigma Chemical Co., St. Louis, Missouri), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (Sigma Chemical Co.), apoptotic activity and tritiated thymidine incorporation (Dupont NEN, Boston, Massachusetts) assays. Normal human bladder smooth muscle cells were cultured with the extracts of the biomaterials or cultured in direct contact with the biomaterials. RESULTS: All naturally derived and synthetic biomaterials tested in this study except alginate exhibited nontoxic and bioactive effects on human bladder smooth muscle cells (HBSMCs) in vitro, as indicated by the 4 types of biocompatibility assays using the extract and direct contact methods. Cell viability, apoptotic properties, metabolic activity and DNA synthesis activity of HBSMCs cultured with the extracts of the biomaterials or cultured in direct contact with the biomaterials were not significantly different from those of negative controls (fresh medium with no extracts or tissue culture plates without biomaterials). CONCLUSIONS: All naturally derived and synthetic biomaterials tested in this study except alginate exhibited nontoxic and bioactive effects on HBSMCs in vitro. This normal primary human bladder smooth muscle cell culture model is suitable for in vitro biocompatibility assessment. It provides information on cell-biomaterial interactions and on the ability of biomaterials to support bioactive cell functions. PMID- 11912451 TI - Use of absorbable cyanoacrylate glue to repair an open cystotomy. AB - PURPOSE: A biodegradable cyanoacrylate glue was tested for its ability to close bladder injuries in an established porcine model. Inflammation and encrustation associated with this glue were examined in a rabbit model. MATERIALS AND METHODS: Four domestic pigs underwent transverse cystotomy, which was closed with absorbable cyanoacrylate glue. Four weeks later the bladder was distended with normal saline to evaluate the repair. A total of 45 rabbits underwent cystotomy, which was closed with polyglactin suture, absorbable cyanoacrylate glue or nonabsorbable 2-octyl cyanoacrylate glue. The bladder was harvested at 4 or 12 weeks to evaluate inflammation, microcalcification and encrustation. RESULTS: All 4 pig bladders tolerated a pressure of 200 mm. Hg 4 weeks after closure. In the rabbit bladders there was no difference in inflammation in the groups at 4 and 12 weeks. The absorbable glue and suture groups had less microcalcification than the 2-octyl cyanoacrylate glue group at 4 and 12 weeks (p = 0.01 and 0.02, respectively). Encrustation was less in the suture and absorbable glue groups than in the 2-octyl cyanoacrylate glue group at 4 and 12 weeks (p = 0.004 and 0.02, respectively). CONCLUSIONS: An experimental absorbable cyanoacrylate glue has the strength to seal a large cystotomy. The inflammatory response to absorbable glue is similar to that to suture at 12 weeks. Absorbable glue does not promote calcification. These properties may make it a suitable material for replacing or augmenting suture in the urinary tract. PMID- 11912452 TI - Norepinephrine content and adrenoceptor function in the bladder of cats with feline interstitial cystitis. AB - PURPOSE: Interstitial cystitis is a chronic syndrome affecting humans and domestic animals, including cats (feline interstitial cystitis). The aggravation of interstitial cystitis symptoms by stress suggests involvement of the sympathetic nervous system. Studies have identified increased sympathetic nervous system activity in patients with interstitial cystitis but to our knowledge effects on bladder function have not been reported. To address this question we measured bladder norepinephrine (NE) content, the electrical field stimulation flux of NE and acetylcholine (ACh), and the effects of feline interstitial cystitis on adrenoceptor (AR) mediated bladder strip contractility. MATERIALS AND METHODS: Bladders were obtained from healthy cats and cats with feline interstitial cystitis. In experiment 1 bladder tissue NE content was determined and the simultaneous release of 3H-NE and 14C-ACh in perfusion bath effluent after electrical field stimulation was measured. NE and ACh release was calculated from the area under the efflux curve. In experiment 2 electrical field stimulation induced contractility of bladder body strips was measured in the presence of 100 nM. to 25 microM. NE only or combined with atipamezole (an alpha2 AR antagonist), propranolol (a beta-AR antagonist) or phentolamine (an alpha-AR antagonist). Antagonists were added to the bath at least 15 minutes before stimulation, after which NE was added in cumulative doses and dose response curves were constructed. RESULTS: Significant increases in NE content and efflux in the absence of alterations in ACh efflux were identified. In the bladder strip studies decreased alpha2 and beta1-AR function was found in strips from cats with feline interstitial cystitis, whereas beta3 or atypical beta-ARs were tentatively identified. CONCLUSIONS: These results support and extend previous studies by identifying an effect of increased sympathetic activity on bladder function in cats with feline interstitial cystitis. PMID- 11912454 TI - Role of supraspinal alpha1-adrenoceptors for voiding in conscious rats with and without bladder outlet obstruction. AB - PURPOSE: Previous studies have shown that spinal alpha1-adrenoceptors can influence voiding in normal rats and in rats with outlet obstruction. Also, at the supraspinal level such receptors may be involved in voiding control. Therefore, we studied in rats the effects on cystometrography of intracerebroventricular administered alpha1-adrenoceptor antagonists. MATERIALS AND METHODS: Continuous cystometry was performed in conscious, freely moving rats with and without bladder outlet obstruction. Cystometric parameters were evaluated before and after intracerebroventricular drug administration. RESULTS: In normal rats intracerebroventricular administration of 8 nmol. kg.-1 prazosin (Pfizer Central Research, Sandwich, United Kingdom) or terazosin (Abbott Laboratories, Abbott Park, Illinois) (nonsubtype selective) caused no change in cystometric parameters. At 24 or 80 nmol. kg.-1 the 2 drugs significantly decreased voiding pressure and increased bladder capacity, voided volume and post void residual urine volume. Administering vehicle had no effect. In rats with outlet obstruction the drug effects were significantly more pronounced than in normal animals (p <0.05), and urinary retention was produced in 50% of rats receiving prazosin. In normal rats the selective alpha1A-adrenoceptor antagonists KMD 3213 (0.8, 8 and 24 nmol. kg.-1) dose dependently depressed voiding pressure, and increased bladder capacity and voided volume, whereas BMY 7378 (selective for alpha1D-adrenoceptors) and A322312 (selective for alpha1B-adrenoceptors) at doses up to 80 nmol. kg.-1 had no effect. CONCLUSIONS: The results suggest that in normal rats and in rats with outflow obstruction volume induced bladder activity involves supraspinal alpha1-adrenoceptors. Bladder outlet obstruction seems to enhance the importance of these receptors. At least in normal rats the alpha1A adrenoceptor subtype seems to mediate the effect. PMID- 11912453 TI - Potassium channel KV alpha1 subunit expression and function in human detrusor muscle. AB - PURPOSE: We determined whether voltage gated K+ (KV) channels are expressed and functional in human detrusor smooth muscle MATERIALS AND METHODS: Information on KV channels was obtained using electrophysiological patch clamp, immunofluorescence, Western blot and isometric tension recording techniques RESULTS: Patch clamp recordings from detrusor cells revealed a Ca2+ independent K+ current that was activated by depolarization in a voltage range near the resting potential of detrusor smooth muscle. The current was inhibited by 3,4 diaminopyridine, a blocker of KV channels. Antibodies targeted to KValpha1 subunits revealed KV1.3 and KV1.6 expression in whole bladder tissue samples and specifically in detrusor smooth muscle cells. New specific blockers of KValpha1 channel currents (correolide and recombinant agitoxin-2) had a myogenic effect, characterized by increased amplitude of spontaneous contractions without an effect on the frequency of contractions or on resting baseline tension. CONCLUSIONS: KValpha1 subunits are expressed and functionally important in human detrusor muscle. PMID- 11912455 TI - Carbon monoxide relaxes the female pig urethra as effectively as nitric oxide in the presence of YC-1. AB - PURPOSE: Nitric oxide (NO) and carbon monoxide (CO) have been suggested to relax smooth muscle by activating soluble guanylate cyclase (sGC), binding to the same site of the enzyme. 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1) (Cayman Co., Malmo, Sweden) increases the catalytic rate of sGC by binding to an allosteric site. We investigated whether YC-1 can modulate the relaxant responses of isolated urethral smooth muscle to exogenous CO, (NO) and electrical field stimulation. MATERIALS AND METHODS: In spontaneously active and noradrenaline (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) pre-contracted preparations of circular urethral smooth muscle from female pigs relaxant responses were evoked by electrical field stimulation before and after incubation with 10(-5) M. YC-1. The concentration-response curves for CO and NO were investigated in noradrenaline pre-contracted strips before and after incubation with YC-1. The tissue contents of cyclic 3',5'-guanosine monophosphate (cGMP) and cyclic adenosine monophosphate after electrical field stimulation, and the administration of CO or NO was investigated in the absence and presence of YC-1. RESULTS: YC-1 significantly increased the amplitude of the relaxations evoked by electrical field stimulation, CO and NO, and simultaneously caused significant increases in the cGMP content in all preparations. The effect on CO induced relaxant responses was conspicuous. In the presence of YC-1 the potency and maximal relaxant effect of CO were similar to those of NO in the absence of YC-1. CONCLUSIONS: YC-1 enhances cGMP dependent relaxant responses of the female pig urethra in vitro. The finding that the response to CO was greatly increased after sensitizing sGC suggests a potential for CO as a relaxant mediator in urethral smooth muscle. PMID- 11912456 TI - Androgen induced norepinephrine release from postganglionic neurons mediates accessory sex organ smooth muscle proliferation. AB - PURPOSE: Guinea pig seminal vesicle smooth muscle displays an initial androgen dependent, proliferative response during early puberty, followed by progression to an androgen resistant, amitotic state in adults. We determined the role of norepinephrine in androgen dependent pubertal proliferation and in the subsequent terminal differentiation of adult seminal vesicle smooth muscle. MATERIALS AND METHODS: Guinea pig seminal vesicle provided a suitable model since its unique anatomy allowed clean harvest of smooth muscle without epithelium. Norepinephrine release from postganglionic adrenergic nerve terminals in seminal vesicle smooth muscle was measured using several techniques. Prazosin sensitive electrical field stimulation of contractile responses qualitatively assessed norepinephrine release. Norepinephrine release was quantified directly in vitro from incubated seminal vesicle smooth muscle minces and indirectly ex vivo from intact tissue using the endogenous seminal vesicle smooth muscle concentration ratio of 3,4 dihydroxyphenylglycol-to-norepinephrine (Sigma Chemical Co., St. Louis, Missouri). Norepinephrine mediated seminal vesicle smooth muscle proliferation was assessed by the time course relationships of androgen induced norepinephrine release, protein kinase C activation-depletion and increases in total DNA, the impact of in vivo reserpine induced norepinephrine depletion on protein kinase C activation-depletion and the mitogenic response, and the alpha1-adrenoceptor mediated mitogenic response in cultured seminal vesicle smooth muscle cells. RESULTS: In prepubertal smooth muscle androgen induced norepinephrine release from postganglionic neurons. The effect was independent of preganglionic innervation. Increased norepinephrine release was concurrent with the onset of androgen induced protein kinase C activation-depletion and cellular proliferation. In vivo norepinephrine depletion to 1% or less of control values by chronic reserpine treatment selectively antagonized the androgen induced increases in smooth muscle DNA and protein kinase C down-regulation. Norepinephrine depletion by reserpine neither induced apoptosis nor altered cell number. Cell culture experiments demonstrated that alpha1-adrenoceptors mediated the proliferative response to norepinephrine. Together these findings indicate that increased norepinephrine release has an obligatory role in androgen dependent muscle cell proliferation during puberty. Terminally differentiated smooth muscle in adults was characterized by androgen resistance to elevated norepinephrine release and protein kinase C activation. CONCLUSIONS: Androgen induced norepinephrine release from postganglionic neurons in seminal vesicle smooth muscle mediated the proliferative response that occurs in early pubertal development. Normal uncoupling of elevated norepinephrine release and protein kinase C activation-depletion may represent a key event in the normal terminal differentiation of accessory sex organ smooth muscle in adults. PMID- 11912458 TI - Epidemiology of anorectal chlamydial and gonococcal infections among men having sex with men in Seattle: utilizing serovar and auxotype strain typing. AB - BACKGROUND: Bacterial sexually transmitted diseases (STDs) among men who have sex with men (MSM) have recently increased in Seattle. GOALS: Serovar and auxotype typing of strains was used to assess the epidemiology of anorectal chlamydial and gonococcal infections among MSM attending an STD clinic. STUDY DESIGN: The prevalences of anorectal chlamydial infection and gonorrhea among MSM attending an STD clinic during the period of 1994 to 1996 were compared with prevalences during 1997 to 1999. A retrospective case-control study of MSM attending an STD clinic between 1997 and 1999 was performed. Anorectal chlamydial isolates were characterized by serovar and gonococcal isolates were characterized by serovar and auxotype. Infected MSM were mapped by residence and strain type. RESULTS: Prevalences of anorectal chlamydial and gonococcal infections increased from 4.0% and 6.3%, respectively, during 1994-1996 to 7.6% and 8.7%, respectively, during 1997-1999 (P = 0.004 and P = 0.013 for chlamydial infection and gonorrhea, respectively). Most chlamydial infections were caused by serovars G (47.9%) and D (29.6%), and most gonococcal infections were caused by auxotype/serovar classes Proto/IB-1 (43.3%), Proto/IB-3 (16.5%), and Proto/IB-2 (10.3%). MSM with anorectal chlamydial infection more often had chlamydial urethritis (P = 0.005) and were not white (P = 0.046), in comparison with controls. MSM with anorectal gonorrhea more often had pharyngeal gonorrhea (P < 0.001), had a history of gonorrhea (P = 0.003), and were younger than age 30 years (P = 0.039), in comparison with controls. Residences of MSM with anorectal gonorrhea were clustered in urban areas, whereas those of MSM with anorectal chlamydial infection were more dispersed. CONCLUSION: Prevalences of anorectal chlamydial infection and gonorrhea among MSM in Seattle have increased dramatically over the past 3 years. Serovar and auxotype analyses indicate these increases are not clonal but are due to the spread of unique distributions of strains that differ from those causing urogenital infections in the same community. PMID- 11912459 TI - Persistence of Chlamydia trachomatis infection detected by polymerase chain reaction in untreated patients. AB - BACKGROUND: Prior studies have used Chlamydia trachomatis culture methods to demonstrate both persistence and spontaneous clearance of genital C trachomatis infection. OBJECTIVE: To further assess the issue of persistence and spontaneous clearance of C trachomatis infection, untreated men and women were evaluated with repeated polymerase chain reaction (PCR) testing. METHODS: Ninety four untreated patients with a prior positive C trachomatis PCR test returning to the Denver Metro Sexually Transmitted Disease Clinic were retested by PCR. RESULTS: The median and range intervals from initial to follow-up testing were 9.0 (2-112) days for men and 10.0 (2-231) days for women. Repeated PCR tests were positive for 29 of 36 men (80.6%) and 45 of 58 women (77.6%). Persistent PCR positivity did not decrease with a longer testing interval. By multivariate analysis, independent predictors of a persistently positive PCR test included nonwhite ethnicity, an interval of more than 3 days since last sexual encounter before the initial test, and an initial PCR optical density value of greater than or equal to 3.0. CONCLUSIONS: In the absence of treatment, a large majority of patients testing positive for C trachomatis by PCR are likely to remain positive for variable periods of time, increasing the risk of transmission and immune-mediated damage. A low initial optical density value and recent sexual contact may be markers for exposure that does not establish infection. PMID- 11912461 TI - Clustering of seropositivities for sexually transmitted infections. AB - BACKGROUND: Serology for different sexually transmitted infections (STIs) is useful for epidemiologic studies on the spread of STIs in different populations. Studying whether seropositivities for different STIs cluster could be useful, both for development of improved serologic markers of sexual behavior in populations and for understanding how STIs may differ in terms of the dynamics of their spread. GOAL: To evaluate the degree of clustering of different STIs in relation to sexual history. STUDY DESIGN: An age- and sexual history-stratified subsample of 275 women from a survey of healthy Swedish women seeking contraceptive advice was tested for human papillomavirus (HPV) types 6, 11, 16, 18, and 33; Chlamydia trachomatis; herpes simplex virus 2 (HSV-2); and human herpesvirus 8. RESULTS: Significant clustering was observed only for HPV types 6 and 11; for HPV types 16, 18, and 33; and for C trachomatis and HSV-2. The serologic marker that correlated best with lifetime number of sex partners was HPV type 16 (odds ratio [OR], 10.2; 95% CI, 3.8-27.6). The combined serologic marker that correlated most highly with sexual history was joint positivity for HPV types 16 and 33 (OR, 25.5; 95% CI, 5.4-120.4). CONCLUSIONS: The degree of clustering between different STIs varies from nonexistent to strong, implying that different STIs commonly have very different transmission dynamics. Certain combinations of STI serologic tests may be useful in epidemiologic studies for predicting sexual behavior in groups. PMID- 11912460 TI - Barriers to reproductive tract infection (RTI) care among Vietnamese women: implications for RTI control programs. AB - BACKGROUND: Vietnamese women may be especially vulnerable to reproductive tract infections (RTIs) and their biological and social sequelae. Few data are available on the prevalence of and health-seeking behavior for RTIs among women in Vietnam. GOAL: To assess prevalence of RTI symptoms, describe treatment seeking behaviors, and identify barriers to care among Vietnamese women. STUDY DESIGN: A population-based survey was conducted among 1163 Vietnamese women aged 18 years to 49 years. RESULTS: Five hundred seven women (43.6%) reported RTI symptoms in the previous 6 months, including abnormal vaginal discharge (78.3%), lower abdominal pain (46.7%), and genital ulcers (3.6%). Sixty-four percent of these women sought care at some type of medical venue: health station (i.e., government clinic; 24.7%), hospital (15.8%), pharmacy (15.2%), or private doctor (8.1%). The remaining women ignored symptoms (24.8%) or were self-treated (11.4%). In multivariate analysis, stigma associated with sexually transmitted infections (odds ratio [OR] = 1.83; 95% confidence interval [CI] = 1.25-2.70); not seeking informal advice (OR = 2.90; 95% CI = 1.82-4.62); mildness of symptoms (OR = 3.01; 95% CI = 1.45-6.23); absence of perceived morbidity (OR = 3.56; 95% CI = 2.20-5.77); and short duration of symptoms (OR = 2.53; 95% CI = 1.04-6.16) were significantly associated with ignoring RTI symptoms. CONCLUSION: A substantial number of women in northern Vietnam who reported RTI symptoms did not seek care. Interventions to raise awareness about RTI symptoms and their consequences, dissipate negative stereotypes, and encourage open discussion about RTIs should facilitate appropriate care-seeking for RTIs. PMID- 11912462 TI - Premarital sexual activities among students in a university in Beijing, China. AB - BACKGROUND: Sexually transmitted diseases are becoming a serious public health problem in China. College students are recognized as one of the age groups most affected. GOAL: The goal was to investigate premarital sexual activities and condom use among college students in Beijing in order to collect the information necessary for research on interventions. STUDY DESIGN: A cross-sectional survey was conducted in June 1999 among students at a university in Beijing. A self administered, anonymous, structured questionnaire was used. RESULTS: Among those interviewed, 41% reported premarital sexual activities: 28% had kissed partners of the opposite sex, 19% had masturbated, and 12% had engaged in sexual intercourse. Among those who had sexual intercourse, 69% had used condoms. CONCLUSION: Today, more students are engaging in premarital sexual intercourse. Thus, it is necessary to reinforce reproductive health education among college students and provide convenient and optional services that are easily accessed. PMID- 11912463 TI - Screening for Neisseria gonorrhoeae and Chlamydia trachomatis at entertainment venues among men who have sex with men. AB - BACKGROUND: To assess the prevalence and increase awareness of Neisseria gonorrhoeae and Chlamydia trachomatis infections among men who have sex with men, a screening program was conducted at three inner-city homosexual entertainment venues and one community function. STUDY DESIGN: Each venue was accessed twice over a 3-month period between March and June 2000. First-catch urine specimens were collected for analysis by polymerase chain reaction (PCR), and throat swabs were collected for culture and PCR. RESULTS: A total of 202 men were tested during the 7 screening nights, including 16 who were tested more than once. From the 186 men tested for the first time, 184 urine specimens were collected, of which 8 (4.3%; 95% CI, 1.9-7.8%) were PCR-positive for C trachomatis and 1 (0.5%; 95% CI, 0-2.1%) was PCR-positive for N gonorrhoeae. Of the 161 throat swabs collected, none were positive for C trachomatis or N gonorrhoeae. CONCLUSIONS: C trachomatis is a potentially significant pathogen in this population of urban homosexual men. Screening programs such as these are valuable as health-promotion exercises. PMID- 11912464 TI - No evidence of an epidemic of locally acquired heterosexual HIV infection in Norway. AB - BACKGROUND: An early sign of a major heterosexual human immunodeficiency virus (HIV) epidemic will be heterosexual infection acquired from persons who were themselves infected through heterosexual intercourse. GOAL: To test the hypothesis that there is a growing heterosexual epidemic of HIV in Norway. STUDY DESIGN: Data from the mandatory, comprehensive, anonymous HIV case reporting system were analyzed concerning Norwegian residents who had acquired HIV heterosexually and for whom such infections were diagnosed before the year 2001. RESULTS: One hundred fifty-five (71%) of 221 men were infected abroad, whereas 107 (76%) of 140 women were infected in Norway (mainly by drug injectors and immigrants); 23 men and 55 women had been infected in Norway by partners who themselves acquired HIV through heterosexual intercourse (secondary heterosexual transmission). There was a slightly increasing incidence of all heterosexual cases and secondary cases. CONCLUSIONS: Secondary heterosexual HIV transmission remains rare in Norway, and a sustainable epidemic of locally acquired infection seems unlikely in the foreseeable future. The magnitude of the heterosexual epidemic will be strongly influenced by infections acquired abroad. PMID- 11912465 TI - Can behavior change explain increases in the proportion of genital ulcers attributable to herpes in sub-Saharan Africa? A simulation modeling study. AB - BACKGROUND: The proportion of cases of genital ulcer disease attributable to herpes simplex virus type 2 (HSV-2) appears to be increasing in sub-Saharan Africa. GOAL: To assess the contributions of HIV disease and behavioral response to the HIV epidemic to the increasing proportion of genital ulcer disease (GUD) attributable to HSV-2 in sub-Saharan Africa. STUDY DESIGN: Simulations of the transmission dynamics of ulcerative sexually transmitted diseases (STDs) and HIV with use of the model STDSIM. RESULTS: In simulations, 28% of GUD was caused by HSV-2 before a severe HIV epidemic. If HIV disease was assumed to double the duration and frequency of HSV-2 recurrences, this proportion rose to 35% by year 2000. If stronger effects of HIV were assumed, this proportion rose further, but because of increased HSV-2 transmission this would shift the peak in HSV-2 seroprevalence to an unrealistically young age. A simulated 25% reduction in partner-change rates increased the proportion of GUD caused by HSV-2 to 56%, following relatively large decreases in chancroid and syphilis. CONCLUSION: Behavioral change may make an important contribution to relative increases in genital herpes. PMID- 11912466 TI - Condom use and sexual behaviors among individuals procuring free male condoms in South Africa: a prospective study. AB - BACKGROUND: Although the South African government has increased the number of male condoms distributed free to the public, there is no understanding of whether these are being used effectively to prevent the spread of sexually transmitted diseases, including HIV infection. GOAL: The study goal was to determine sexual behaviors and barriers to condom use among individuals procuring condoms distributed free to the public sector. STUDY DESIGN: In a prospective study, individuals who procured condoms from 12 public health facilities across South Africa were recruited and observed for the next 5 weeks. RESULTS: The 384 successfully observed subjects reported 3262 sexual contacts, of which 2637 (81%) involved protection with a condom. In multivariate analysis, alcohol consumption and use of other contraceptives were associated with unprotected sexual intercourse. CONCLUSION: The levels of condom use are relatively high in this population, but these data highlight several important barriers to condom use that may be targeted by future interventions. PMID- 11912467 TI - Surveillance of antibiotic resistance of Neisseria gonorrhoeae isolates in China, 1993-1998. AB - BACKGROUND: The prevalence of sexually transmitted diseases (STDs) has been increasing in China since the 1980s. Because gonorrhea is the most frequently reported STD there, information on the antimicrobial susceptibility of Neisseria gonorrhoeae will aid in its control. GOAL: To investigate the antimicrobial susceptibility of N gonorrhoeae isolates in China and to provide data for formulation of treatment guidelines and control policies. METHODS: The agar dilution technique was used to determine antimicrobial susceptibility, and acidimetric method was used to test for penicillinase-producing N gonorrhoeae. RESULTS: A total of 3186 gonococcal isolates were tested during the 6-year study period. The rate of resistance to penicillin was 66.70%; 8.14% of isolates were penicillinase-producing N gonorrhoeae. The percentage of tetracycline-resistant isolates was 92.03%, and that of highly tetracycline-resistant isolates was 4.65%. The rate of resistance to ciprofloxacin was also relatively high (34.25%). The rates of resistance to spectinomycin and ceftriaxone were 0.44% and 0.57%. CONCLUSIONS: The gonococcal isolates in China are relatively highly resistant to penicillin, tetracycline, and ciprofloxacin, but most of them are still susceptible to spectinomycin and ceftriaxone. Standardized treatment of gonorrhea is needed to prevent further spread of resistant gonococcal strains. PMID- 11912469 TI - Proposal of a clinico-histological "glioma score" as a prognostic index in high grade glioma patients. Preliminary observations in a series of 80 cases. AB - BACKGROUND: Despite of several multimodal treatments, malignant gliomas still have a poor outcome. In order to identify subgroups of patients with different prognosis, we propose a clinical and histological score (GS). METHODS: Eighty consecutive patients operated on for a high-grade glioma and treated with adjuvant therapy entered the study. In relation to age at diagnosis, preoperative Karnofsky Performance Status (KPS), and MIB-1 index, patients have been splitted in 4 groups (GS 0-III). RESULTS: The overall mean survival of the entire cohort was 18.2 months (median 12). Patients with GS 0 have a mean survival rate of 30.0 months, with GS I 23.1 months, with GS II 12.1 months, and with GS III 9.0 months (p=0.0001). Moreover, mean survival with a KPS = or >70 was 29.0 in GS 0, 26.0 in GS I, 10.0 in GS II, and 0 in GS III patients (p<0.0001). CONCLUSIONS: On the basis of these preliminary observations, we discuss the utility of our "glioma score" as a prognostic indicator for patients operated on for cerebral malignant gliomas and treated postoperatively with adjuvant therapy. PMID- 11912468 TI - Primary care physician attitudes regarding sexually transmitted diseases. AB - BACKGROUND: Primary care physicians see the majority of patients with sexually transmitted diseases (STDs), but little is known about their attitudes regarding STD-related issues. GOAL: The study goal was to determine the attitudes of primary care physicians toward STD-related issues, to determine physicians' characteristics associated with attitudes, and to examine the relationship of attitudes to STD counseling practices. STUDY DESIGN: A cross-sectional survey was mailed to randomly selected primary care physicians in Pennsylvania. RESULTS: Of 1054 eligible physicians, 541 (51%) responded. Although most physicians were comfortable discussing sex-related issues with their patients (89%), many believed their STD counseling was ineffective (70%), their medical school STD training was inadequate (48%), or that they were not responsible for STD preventive services for their patients (43%). Overall, STD-related attitudes were more positive among physicians who were female, worked in clinic settings, and received adequate training in STDs. More positive attitude scores were significantly associated with performance of six specific risk-assessment and counseling behaviors. CONCLUSIONS: Many physicians reported low confidence, limited responsibility, and time barriers that may affect their STD-prevention practices. Interventions that influence STD-related attitudes may improve STD prevention practices by primary care physicians. PMID- 11912470 TI - Instrumentation for posterior stabilisation of cervical traumatic and degenerative disorders: bullet-shaped implant and titanium cable. AB - BACKGROUND: Numerous implants for posterior stabilisation of cervical spine have been described so far. The aims of all these implants and techniques are rigid spinal stabilisation without neurologic damage, restoration of neuroanatomy and excellent radiological studies in postoperative period. The objective of this study was to determine the effectively and clinical safety of this system. METHODS: We conducted a retrospective analysis of patients treated with posterior stabilisation system for the stabilisation of traumatic and osteodegerative disorders of lower cervical spine in our department. This posterior cervical stabilisation system consist of titanium bullet-shaped implant (Ti-Frame) and titanium cables (sof' wire). RESULTS: All patients underwent only posterior fixation except 2 (anterior decompression and posterior stabilisation in 2 stages) and postoperative early immobilisation was allowed with Philadelphia collar in all patients. At the follow-up period 15.2 months (9-25 months), none of the patients had superficial or deep infection, implant resection or failure. CONCLUSIONS: In conclusion, this system (Ti-frame and titanium cables) is a simple, safe and effective system for posterior cervical stabilisation in patients with traumatic and osteodegenerative disorders due to provide rigid fixation and allow CT and MR imaging without the significant artifact. PMID- 11912471 TI - Carotid endarterectomy: a retrospective analysis. Microendarterectomy and transcranial Doppler ultrasound monitoring. AB - BACKGROUND: Analyses of the results obtained following our protocol for the surgical therapy of carotid artery stenosis is the aim of this study. METHODS: Ninety-one procedures were performed on 84 patients by the same surgical team. Indications for carotid endarterectomy (CEA) were 50-70% internal carotid artery (ICA) stenosis in 8% of the patients (100% symptomatic), 70-90% stenosis in 67% (64% symptomatic) and 90-99% ICA stenosis in 25% of our patients (33% symptomatic). Carotid shunt during arterial cross-clamp was only used in 4 cases (4.4%) following trans-cranial Doppler ultrasound (TCD) intraoperative monitoring findings. Operative microscope or surgical loupes were used for the complete removal of atheroma and the direct suture of the arteriotomy. Heparin was only antagonised in selected cases. RESULTS: No mortality and no permanent deficit related to ischemic events were present in our patients; a transient neurological deficit occurred in 1 case (1.1%). Two patients died (2.2%): 1 due to myocardial infarction and the second one to bronchopneumonia. Transient peripheral nerve deficits were noted in 2 cases. CONCLUSIONS: In order to reduce to a minimum ischemic risks during CEA, our preliminary experience advises microsurgical technique for the complete removal of the atheroma and suture, TCD evaluation of collateral reserve during cross-clamping, and the use of randomised multicenter studies guidelines for the selection of patients. More attention must also be given to postoperative care, since hemodynamic instability represents the main risk for our patients. Overnight intensive care unit (ICU) surveillance in all patients may help to avoid fatal complications. PMID- 11912472 TI - Cervical spinal cord intramedullary abscess. Case report. AB - A case of progressive symptoms and signs of cervical spinal cord damage due to intramedullary abscess is reported. The literature is reviewed and the radiological features, particularly magnetic resonance image, are analyzed. PMID- 11912473 TI - Pigment villonodular synovitis of the spine. Case report and review of the literature. AB - Pigmented villonodular synovitis (PVNS) is a disease of the joints which uncommonly involves the spine. We present a 70-year-old woman with radicular symptoms who was found to have a mass arising from a lumbar zygapophyseal joint with extension into the spinal canal. Following gross-total excision of the mass, histology revealed PVNS. One month after surgery, the patient had no symptoms and there was no evidence of residual or recurrent disease. PMID- 11912474 TI - Spinal intradural endodermal cyst located anterior to the cervical spinal cord. AB - Spinal endodermic cysts are rare and may be associated with other congenital anomalies (mediastinal cysts, bony defects of the vertebral body), derive from defective displacement of the endoderm of the intestinal tract or, more rarely, the respiratory one. The authors describe a case of endodermic cyst of the cervical spine localized anteriorly to the spinal cord. In the case we treated the patient was a 17-year-old male who presented hypostenia of the lower limbs accompanied by hyperreflexia and spasticity; physical examination was negative. MRI, CT and CT-myelography documented a well-defined mass situated anteriorly to the spinal cord at C2-C4 level. A total-body CT-scan excluded the existence of other malformations. Surgery was performed via an anterior approach and the cyst completely removed. Histological examination confirmed the endodermal nature of the cyst. At long-term follow-up examination the patient was neurologically intact. Postoperative MRI showed a small residue of the cyst wall without spinal cord compression. Endodermic spinal cysts are very rare intradural lesions, predominantly with an anterior cervical localization, which derive from misplaced embryonic and endodermic cells. In the majority of cases, clinical onset is insidious and has a discontinuous progression. The best diagnostic tool is MRI. Treatment of choice is total surgical removal or emptying of the cyst followed by fenestration. PMID- 11912475 TI - Post-traumatic intradiploic epidermoid cyst. A case report. AB - A patient with an intradiploic epidermoid cyst of calvarium was presented. Head injury that may cause the inclusion of epidermal cells into the diploe of the bone was present in his medical history. Skull radiographs and cranial magnetic resonance imaging (MRI) demonstrated the tumor. The tumor was totally removed and the patient was discharged free of symptoms. PMID- 11912476 TI - Cerebellar metastasis from clear cell sarcoma of the kidney. A case report with immunohistochemistry. AB - The aim of this study was to describe a child with a right cerebellar hemisphere metastasis from primary clear cell sarcoma of the kidney without evidence of bone metastases, and to investigate the immunohistochemical features of primary and metastatic tumors. A 12-month old boy was admitted our hospital due to an abdominal mass. Abdominal computed tomography revealed a large right renal tumor. Tumor was removed with nephrectomy. Histopathologic examination of tumor revealed clear cell sarcoma of the kidney. The patient received radiotherapy and chemotherapy in postoperative period. He suffered from gait disturbance and confusion 8 months later. A computed tomography scan revealed a tumor that was enhanced with contrast medium at right cerebellar hemisphere concomitant with ventricular enlargement. After ventriculo-peritoneal shunting procedure, tumor was excised totally and histopathologic diagnosis showed metastasis of clear cell sarcoma of the kidney. Immunohistochemically vimentin, actin, desmin, neuron specific enolase, cytokeratin, P 53, Ki-67 and P-170 were performed using formalin fixed, paraffin embedded sections. Both of the tumors were positive for vimentin and negative for desmin, actin, neuron specific enolase, cytokeratin and P 53. Scattered nuclei were stained by Ki-67 in primary and metastatic cerebellar tumor. Both primary and metastatic tumors were negative for p53 and P-170. The treatment consisted of surgery, radiotherapy and chemotherapy. The patient is alive and well without evidence of recurrence 16 months after second surgery. Clear cell sarcoma of the kidney is most commonly associated with bone metastasis. Cerebellar metastasis of clear cell sarcoma of the kidney is very unusual. To the best of our knowledge, this patient is second case in the English literature. With review of the literature, our immunohistochemical findings support the theory that relapse and metastasis of primary clear cell sarcoma of the kidney are not related with increase of aggressiveness. PMID- 11912477 TI - Middle cerebral artery duplication associated with multiple intracranial aneurysms. Case report. AB - We present a case of middle cerebral artery (MCA) duplication associated with ipsilateral distal MCA and contralateral MCA bifurcation aneurysms. PMID- 11912478 TI - Primary leptomeningeal melanomatosis: early leptomeningeal enhancement on MRI. AB - One unusual case of primary leptomeningeal melanomatosis is presented. The patient, 53-year-old male, was admitted to our observation for 1 month history of psychasthenia and amnesia. Despite several polyspecialistic clinical, neuroradiological and cytological examinations, conclusive diagnosis was made only with a biopsy of leptomeningeal nodule. The present case allows as to identify 2 stages in the evolution of primary diffuse leptomeningeal melanomatosis. The initial phase is characterized by slight mental impairment without hydrocephalus; during the later phase there is severe, diffuse neurological impairment and both CT and MRI show hypercaptation of the intracranial leptomeninges and multiple, leptomeningeal tumoral nodules. PMID- 11912479 TI - Thallium-201 SPECT and equivocal neuroradiological supratentorial lesions. PMID- 11912480 TI - [Daily dialysis: evaluation of the first year of experience at home and in a limited care center]. AB - BACKGROUND: Among self dialysis treatments, daily dialysis is encountering a growing interest. Aim of this study was to evaluate results of the first year of daily dialysis in our Center. METHODS: Since November 1998, twelve patients started daily dialysis. One patient started RRT on daily dialysis; one patient was in training; 8 were on home dialysis, 3 in the limited care center. Selection of patients was performed according to wide acceptance criteria as for age (range 33-61 years), dialysis follow-up (range 1-23 years), comorbidity (=/>1 comorbid factor present in 8). Dialysis schedule consisted of 6 sessions per week (2-3 hours), blood flow 250-320 ml/min, individualized dialysate. Occasional shift to 3-4 times per week were allowed for logistic or working reasons. RESULTS: Results were analyzed taking into account patient satisfaction and main clinical parameters. In 9/12 the choice of treatment resulted from both clinical reasons and patient preferences, while in 3 was due to clinical indications (1/3 dropped out). The main reasons of choice were logistic or research of the best treatment. The most common fears regarded fistula and needle puncturing. Despite the time unconvenience, the rapidly regained well being was the reason for choosing this treatment. Also in this relatively short follow-up the favorable results reported as for weight gain, blood pressure control and metabolic pattern are confirmed. The few side effects were multifactorial (fistula thrombosis after blood pressure normalization, 2 recurrences of atrial fibrillation). CONCLUSIONS: In conclusion, daily dialysis resulted also in our centre as a promising alternative even in difficult patients. PMID- 11912481 TI - The use of the small caliber JJ stent with anti-reflux valve in double kidney transplant. Personal experience. AB - BACKGROUND: In the experience of other authors, double kidney transplant have a higher complication rate (30%) if compared with single renal graft. In personal experience the use of small calibre ureteral stents with antireflux valve can reduce this complication rate. METHODS: From November 1999 to April 2001, at the A.S.O. S. Giovanni Battista in Turin, we performed 29 double kidney transplantations with the application of small calibre stents in 20 male and 9 female patients, aged 50-74 years. The uretero-neocystostomies were carried out according to Lich-Gregoire technique, and the JJ stents used were pediatric ones, 12 cm long and 4.8 Ch, with antireflux valve. RESULTS: We complained only 2 urological complications out of 58 anastomoses (distal unilateral ureteral necrosis in 1 case and total ureteral necrosis in the other). CONCLUSIONS: Complication rate is lower than in the literature: the authors suggest that the use of small calibre JJ stents can keep the complication rate low in double kidney transplant. PMID- 11912482 TI - [Stenosis of the new bladder neck, a complication of radical prostatectomy: personal experience]. AB - BACKGROUND: To describe personal experience on stenosis of the new bladder neck, a complication of radical prostatectomy. After a review of the literature, guidelines are proposed to avoid this complication, both from a prophylactic and treatment point of view. METHODS: Between 1992 and 2000, 54 patients were submitted to radical prostatectomy, 18 of whom later presented stenosis in the new bladder neck. Mean age of patients was 65.6 years (range 55-76). According to TNM classification, 46 patients (85%) were stage pT2N0M0 and eight patients (15%) stage pT3N0M0, no significant correlation being found with PSA values. All patients were submitted to pre- and postmicturition retrograde cystourethrography, four weeks after surgery; patients presenting dysuric symptoms also underwent further retrograde cystourethrography and flowmeter examinations. RESULTS: Of the 54 patients, 18 (33%) presented stenosis of the new bladder neck as a complication of radical prostatectomy. Of these 18 patients, 12 (22%) presented acute dysuria symptoms. In 14 cases, one-two cycles of urethral dilation were sufficient to improve flowmeter values. In the four remaining cases showing no improvement following urethral dilations, endoscopic resection was necessary in two and urethrotomy according to Sachse in the other two. CONCLUSIONS: Stenosis of the anastomosis of the new bladder neck is a complication of radical prostatectomy occurring within six months of surgery, no correlation being found with tumour stage, recurrence, or duration of catheter in situ. Of the 18 patients presenting stenosis in the series described, in 78%, cycles of urethral dilation were sufficient to successfully resolve the complication thus avoiding further surgery, which, on the other hand, was necessary in four patients, two submitted to endoscopic resection of the stenosis and the other two to incision according to Sachse. PMID- 11912483 TI - Management of membranous nephropathy. AB - The management of membranous nephropathy first requires the recognition of whether the disorder is primary (idiopathic) or secondary. Next, a familiarity with its natural history and knowledge of our current capacity to predict those patients with the worst outcome is reviewed. Treatment options of those at risk of progression with immunosuppressive drugs is then discussed along with the required accompaniment of risk reduction strategies including idealization of the blood pressure, the use of angiotensin enzyme inhibitor therapy and perhaps dietary protein restriction. As well treatment directed at other complications of the disease process including the hyperlipidemia and hypercoagulability are considered as part of the management to reduce risks. Lastly, therapies directed towards preventing or reducing the complications of immunosuppressive drugs such as trimethoprim-sulfamethoxazole prophylaxis against pneumocystis carini pneumonia and biphosphonates to reduce bone mass loss from corticosteroid therapy are discussed. PMID- 11912484 TI - Diagnosis and treatment of atherosclerotic renal artery stenosis (ARAS). AB - Atherosclerotic renal artery stenosis (ARAS) is an increasingly common cause of secondary hypertension and progressive chronic renal failure. Recent studies provide valuable information about the pathophysiology, natural history, diagnosis and treatment of ARAS. The pathophysiology of ARAS is more complex than experimental models using clipped renal arteries because the renal artery narrowing is gradual, may be bilateral, may affect smaller intra-renal arteries and other co-existing nephropathies are often present. Patients with ARAS have high mortality due to associated co-morbidity and progression of renal failure may be less common than previously thought. Magnetic resonance arteriography offers great promise for diagnosing of ARAS as it is non-invasive and can provide data on kidney function. In patients with ARAS, the co-existence of atherosclerotic disease in other vascular beds means that aspirin, blood pressure reduction, advice to stop smoking and lipid lowering therapy are likely to be associated with reduced vascular events. The effect of these approaches on the progression of ARAS is unclear but likely to be beneficial. Re-vascularisation of occluded renal arteries is an attractive option for treatment of ARAS but data from the few randomised controlled studies that have been published do not support its widespread application. Arterial stenting has a higher technical success rate than angioplasty while surgical revascularisation does not appear to improve outcome compared with angioplasty. Recent studies examining functional and histological features of kidneys supplied by atherosclerotic stenosed renal arteries may explain why revascularisation is not always beneficial. The results of on-going studies may identify sub-groups of patients with ARAS who gain a clear benefit from re-vascularisation. In the meantime it seems reasonable to attempt re-vascularisation in the following circumstances: severe hypertension resistant to medical therapy, rapidly progressive renal failure with no obvious cause other than ARAS and recurrent flash pulmonary oedema. PMID- 11912485 TI - Hypertension in end-stage renal disease. AB - Chronic renal failure is common. Recent estimates from the United States suggest that one in 10 adults has an elevated serum creatinine. Hypertension and renal disease are intimately connected at many levels, and clearly accelerate each other s course. Hypertension is an almost universal feature of end-stage renal disease, a state of frightening cardiovascular risk. Surprisingly, most recent observational studies have shown an association between low blood pressure and increased mortality, a result that may engender therapeutic nihilism in the absence of large randomised trials. This observation may be due to reverse causality, as the age and cardiovascular comorbidity of patients reaching end stage renal disease is considerable. When outcomes other than death are considered, especially progressive left ventricular hypertrophy, but also ischaemic heart disease and congestive heart failure, more predictable and expected associations are seen, with rising blood pressure appearing to be a deleterious parameter. Uraemia appears to be a state of premature senescence, and arterial rigidity, whose clinical corollary is wide pulse pressure, is a characteristic feature. Recent observational studies have focused on pulse pressure, rather than the traditional approach of analysing its components, systolic and diastolic blood pressure, in isolation. High pulse pressure appears to be a marker of short survival in dialysis patients, but disentangling this association from old age and pre-existing cardiovascular conditions is challenging. Remarkably, and regrettably, no large scale randomised controlled studies examining strategies that tackle the issue of hypertension in dialysis patients have yet to be initiated. PMID- 11912486 TI - Entero-vesical fistulae in surgical practice. AB - Any disease or traumatic event that involving the integrity of the urinary tract or of the bowel, may led to an entero-vesical fistula formation. The clinical onset of this pathology, is described on the basis of usual clinical pictures and of some unusual cases mimicking a bladder cancer. Moreover, the guide lines for a gold standard in the diagnosis and treatment of such pathology are described. PMID- 11912487 TI - Shaggy/GSK3 antagonizes Hedgehog signalling by regulating Cubitus interruptus. AB - The Drosophila protein Shaggy (Sgg, also known as Zeste-white3, Zw3) and its vertebrate orthologue glycogen synthase kinase 3 (GSK3) are inhibitory components of the Wingless (Wg) and Wnt pathways. Here we show that Sgg is also a negative regulator in the Hedgehog (Hh) pathway. In Drosophila, Hh acts both by blocking the proteolytic processing of full-length Cubitus interruptus, Ci (Ci155), to generate a truncated repressor form (Ci75), and by stimulating the activity of accumulated Ci155 (refs 2-6). Loss of sgg gene function results in a cell autonomous accumulation of high levels of Ci155 and the ectopic expression of Hh responsive genes including decapentaplegic (dpp) and wg. Simultaneous removal of sgg and Suppressor of fused, Su(fu), results in wing duplications similar to those caused by ectopic Hh signalling. Ci is phosphorylated by GSK3 after a primed phosphorylation by protein kinase A (PKA), and mutating GSK3 phosphorylation sites in Ci blocks its processing and prevents the production of the repressor form. We propose that Sgg/GSK3 acts in conjunction with PKA to cause hyperphosphorylation of Ci, which targets it for proteolytic processing, and that Hh opposes Ci proteolysis by promoting its dephosphorylation. PMID- 11912488 TI - The Drosophila immune response against Gram-negative bacteria is mediated by a peptidoglycan recognition protein. AB - The antimicrobial defence of Drosophila relies largely on the challenge-induced synthesis of an array of potent antimicrobial peptides by the fat body. The defence against Gram-positive bacteria and natural fungal infections is mediated by the Toll signalling pathway, whereas defence against Gram-negative bacteria is dependent on the Immune deficiency (IMD) pathway. Loss-of-function mutations in either pathway reduce the resistance to corresponding infections. The link between microbial infections and activation of these two pathways has remained elusive. The Toll pathway is activated by Gram-positive bacteria through a circulating Peptidoglycan recognition protein (PGRP-SA). PGRPs appear to be highly conserved from insects to mammals, and the Drosophila genome contains 13 members. Here we report a mutation in a gene coding for a putative transmembrane protein, PGRP-LC, which reduces survival to Gram-negative sepsis but has no effect on the response to Gram-positive bacteria or natural fungal infections. By genetic epistasis, we demonstrate that PGRP-LC acts upstream of the imd gene. The data on PGRP-SA with respect to the response to Gram-positive infections, together with the present report, indicate that the PGRP family has a principal role in sensing microbial infections in Drosophila. PMID- 11912489 TI - Functional genomic analysis of phagocytosis and identification of a Drosophila receptor for E. coli. AB - The recognition and phagocytosis of microbes by macrophages is a principal aspect of innate immunity that is conserved from insects to humans. Drosophila melanogaster has circulating macrophages that phagocytose microbes similarly to mammalian macrophages, suggesting that insect macrophages can be used as a model to study cell-mediated innate immunity. We devised a double-stranded RNA interference-based screen in macrophage-like Drosophila S2 cells, and have defined 34 gene products involved in phagocytosis. These include proteins that participate in haemocyte development, vesicle transport, actin cytoskeleton regulation and a cell surface receptor. This receptor, Peptidoglycan recognition protein LC (PGRP-LC), is involved in phagocytosis of Gram-negative but not Gram positive bacteria. Drosophila humoral immunity also distinguishes between Gram negative and Gram-positive bacteria through the Imd and Toll pathways, respectively; however, a receptor for the Imd pathway has not been identified. Here we show that PGRP-LC is important for antibacterial peptide synthesis induced by Escherichia coli both in vitro and in vivo. Furthermore, totem mutants, which fail to express PGRP-LC, are susceptible to Gram-negative (E. coli), but not Gram-positive, bacterial infection. Our results demonstrate that PGRP-LC is an essential component for recognition and signalling of Gram-negative bacteria. Furthermore, this functional genomic approach is likely to have applications beyond phagocytosis. PMID- 11912490 TI - Dissecting the link between stress fibres and focal adhesions by CALI with EGFP fusion proteins. AB - Chromophore-assisted laser inactivation (CALI) is a light-mediated technique used to selectively inactivate proteins within cells. Here, we demonstrate that GFP can be used as a CALI reagent to locally inactivate proteins in living cells. We show that focused laser irradiation of EGFP-alpha-actinin expressed in Swiss 3T3 fibroblasts results in the detachment of stress fibres from focal adhesions (FAs), whereas the integrity of FAs, as determined by interference reflection microscopy (IRM), is preserved. Moreover, consistent with a function for focal adhesion kinase (FAK) in FA signalling and not FA structure, laser irradiation of EGFP-FAK did not cause either visible FA damage or stress fibre detachment, although in vitro CALI of isolated EGFP-FAK decreased its kinase activity, but not its binding to paxillin. These data indicate that CALI of specific FA components may be used to precisely dissect the functional significance of individual proteins required for the maintenance of this cytoskeletal structure. In vitro CALI experiments also demonstrated a reduction of EGFP-alpha-actinin binding to the cytoplasmic domain of the beta(1) integrin subunit, but not to actin. Thus, alpha-actinin is essential for the binding of microfilaments to integrins in the FA. CALI-induced changes in alpha-actinin result in the breakage of that link and the subsequent retraction of the stress fibre. PMID- 11912491 TI - Nucleotide exchange factor GEF-H1 mediates cross-talk between microtubules and the actin cytoskeleton. AB - Regulation of the actin cytoskeleton by microtubules is mediated by the Rho family GTPases. However, the molecular mechanisms that link microtubule dynamics to Rho GTPases have not, as yet, been identified. Here we show that the Rho guanine nucleotide exchange factor (GEF)-H1 is regulated by an interaction with microtubules. GEF-H1 mutants that are deficient in microtubule binding have higher activity levels than microtubule-bound forms. These mutants also induce Rho-dependent changes in cell morphology and actin organization. Furthermore, drug-induced microtubule depolymerization induces changes in cell morphology and gene expression that are similar to the changes induced by the expression of active forms of GEF-H1. Furthermore, these effects are inhibited by dominant negative versions of GEF-H1. Thus, GEF-H1 links changes in microtubule integrity to Rho-dependent regulation of the actin cytoskeleton. PMID- 11912492 TI - Identification of a link between the tumour suppressor APC and the kinesin superfamily. AB - The tumour suppressor gene adenomatous polyposis coli (APC) is mutated in sporadic and familial colorectal tumours. APC is involved in the proteasome mediated degradation of beta-catenin, through its interaction with beta-catenin, GSK-3 beta and Axin. APC also interacts with the microtubule cytoskeleton and has been localized to clusters near the distal ends of microtubules at the edges of migrating epithelial cells. Moreover, in Xenopus laevis epithelial cells, APC has been shown to move along microtubules and accumulate at their growing plus ends. However, the mechanism of APC accumulation and the nature of these APC clusters remain unknown. We show here that APC interacts with the kinesin superfamily (KIF) 3A-KIF3B proteins, microtubule plus-end-directed motor proteins, through an association with the kinesin superfamily-associated protein 3 (KAP3). The interaction of APC with KAP3 was required for its accumulation in clusters, and mutant APCs derived from cancer cells were unable to accumulate efficiently in clusters. These results suggest that APC and beta-catenin are transported along microtubules by KAP3-KIF3A-KIF3B, accumulate in the tips of membrane protrusions, and may thus regulate cell migration. PMID- 11912493 TI - Cdc25b phosphatase is required for resumption of meiosis during oocyte maturation. AB - In a wide variety of animal species, oocyte maturation is arrested temporarily at prophase of meiosis I (ref. 1). Resumption of meiosis requires activation of cyclin-dependent kinase-1 (CDK1, p34cdc2), one component of maturation-promoting factor (MPF). The dual specificity phosphatases Cdc25a, Cdc25b and Cdc25c are activators of cyclin-dependent kinases; consequently, they are postulated to regulate cell-cycle progression in meiosis and mitosis as well as the DNA-damage response. We generated Cdc25b-deficient (Cdc25b-/-) mice and found that they are viable. As compared with wildtype cells, fibroblasts from Cdc25b-/- mice grew vigorously in culture and arrested normally in response to DNA damage. Female Cdc25b-/- mice were sterile, and Cdc25b-/- oocytes remained arrested at prophase with low MPF activity. Microinjection of wildtype Cdc25b mRNA into Cdc25b-/- oocytes caused activation of MPF and resumption of meiosis. Thus, Cdc25b-/- female mice are sterile because of permanent meiotic arrest resulting from the inability to activate MPF. Cdc25b is therefore essential for meiotic resumption in female mice. Mice lacking Cdc25b provide the first genetic model for studying the mechanisms regulating prophase arrest in vertebrates. PMID- 11912494 TI - Pbx1 inactivation disrupts pancreas development and in Ipf1-deficient mice promotes diabetes mellitus. AB - Pbx1 is a member of the TALE (three-amino acid loop extension) class of homeodomain transcription factors, which are components of hetero-oligomeric protein complexes thought to regulate developmental gene expression and to maintain differentiated cell states. In vitro studies have shown that Pbx1 regulates the activity of Ipf1 (also known as Pdx1), a ParaHox homeodomain transcription factor required for the development and function of the pancreas in mice and humans. To investigate in vivo roles of Pbx1 in pancreatic development and function, we examined pancreatic Pbx1 expression, and morphogenesis, cell differentiation and function in mice deficient for Pbx1. Pbx1-/- embryos had pancreatic hypoplasia and marked defects in exocrine and endocrine cell differentiation prior to death at embryonic day (E) 15 or E16. In these embryos, expression of Isl1 and Atoh5, essential regulators of pancreatic morphogenesis and differentiation, was severely reduced. Pbx1+/- adults had pancreatic islet malformations, impaired glucose tolerance and hypoinsulinemia. Thus, Pbx1 is essential for normal pancreatic development and function. Analysis of trans heterozygous Pbx1+/- Ipf1+/- mice revealed in vivo genetic interactions between Pbx1 and Ipf1 that are essential for postnatal pancreatic function; these mice developed age-dependent overt diabetes mellitus, unlike Pbx1+/- or Ipf1+/- mice. Mutations affecting the Ipf1 protein may promote diabetes mellitus in mice and humans. This study suggests that perturbation of Pbx1 activity may also promote susceptibility to diabetes mellitus. PMID- 11912495 TI - Genetic analysis of the mouse brain proteome. AB - Proteome analysis is a fundamental step in systematic functional genomics. Here we have resolved 8,767 proteins from the mouse brain proteome by large-gel two dimensional electrophoresis. We detected 1,324 polymorphic proteins from the European collaborative interspecific backcross. Of these, we mapped 665 proteins genetically and identified 466 proteins by mass spectrometry. Qualitatively polymorphic proteins, to 96%, reflect changes in conformation and/or mass. Quantitatively polymorphic proteins show a high frequency (73%) of allele specific transmission in codominant heterozygotes. Variations in protein isoforms and protein quantity often mapped to chromosomal positions different from that of the structural gene, indicating that single proteins may act as polygenic traits. Genetic analysis of proteomes may detect the types of polymorphism that are most relevant in disease-association studies. PMID- 11912497 TI - Human Toll-like receptor 4 recognizes host-specific LPS modifications. AB - Lipopolysaccharide (LPS) is the principal proinflammatory component of the Gram negative bacterial envelope and is recognized by the Toll-like receptor 4 (TLR4) MD-2 receptor complex. Bacteria can alter the acylation state of their LPS in response to environmental changes. One opportunistic bacterium, Pseudomonas aeruginosa, synthesizes more highly acylated (hexa-acylated) LPS structures during adaptation to the cystic fibrosis airway. Here we show that human, but not murine, TLR4-MD-2 recognizes this adaptation and transmits robust proinflammatory signals in response to hexa-acylated but not penta-acylated LPS from P. aeruginosa. Whereas responses to lipidIVA and taxol are dependent on murine MD-2, discrimination of P. aeruginosa LPS structures is mediated by an 82-amino-acid region of human TLR4 that is hypervariable across species. Thus, in contrast to mice, humans use TLR4 to recognize a molecular signature of bacterial-host adaptation to modulate the innate immune response. PMID- 11912502 TI - Freezing rate affects the survival of a short-term freezing stress in Panagrolaimus davidi, an Antarctic nematode that survives intracellular freezing. AB - The ability of the Antarctic nematode Panagrolaimus davidi to survive a short term freezing stress depended upon the rate of freezing of its surroundings, measured as the duration of the sample exotherm. The freezing rate increased as the sample volume and freezing temperature decreased and resulted in fewer nematodes surviving. This appears to be due to the greater risk of physical damage by ice crystal growth at high freezing rates. Once frozen the nematodes will then survive exposure to lower temperatures. The environment of the nematode is likely to produce the slow rate of freezing of its surroundings that is necessary for its survival. PMID- 11912503 TI - Effects of the cryopreservation procedures on recovered rice cell populations. AB - Cryogenic storage of plant cells allows the long-term maintenance of valuable genotypes. Cryopreservation of calli and cell suspensions is often performed using cryoprotectants and slow cooling rates. Rice calli (Oryza sativa L.) were cryopreserved by this procedure as well as by direct immersion in liquid nitrogen without cryoprotection. Subsequently, the characteristics of the recovered cells as well as the effects of putative cryoselection were investigated by microscopic observations and flow cytometric analyses. For this purpose, protoplasts were prepared from calli that had been cryopreserved by direct plunging into liquid nitrogen and from their unfrozen controls. Results show that direct immersion in liquid nitrogen of calli pre-treated with abscisic acid is a fast and highly efficient freezing procedure that maintains the main characteristics of the cell populations and appears to increase their metabolic activity PMID- 11912504 TI - Field performance of sugarcane (Saccharum sp.) plants derived from cryopreserved calluses. AB - This study compared the field performance of sugarcane plants originating from three different sources: control, non-cryopreserved embryogenic calluses, cryopreserved embryogenic calluses and macropropagated material of the same commercial hybrid. Several agronomic traits were evaluated on 100 plants per treatment over a 27-month period covering the growth of the stool and of the first ratoon. Significant differences between treatments were observed only during the first six months of field growth of sugarcane stools. Stems produced from in vitro cultured material, irrespective of their cryopreservation status, had a smaller diameter and a shorter height than those produced from macropropagated material. These differences disappeared by 12 months of stool field growth. PMID- 11912505 TI - Effects of cryopreservation on developmental competency, cytological and molecular stability of citrus callus. AB - Cell suspensions of twelve citrus genotypes were successfully cryopreserved by vitrification. All genotypes survived cryopreservation with >90% viability and the surviving cells of some genotypes regenerated somatic embryos better than the controls. Single-cell sibling lines of cultivar Newhall were used for cytological and molecular examination. It was found that the ploidy constitution remained genetically stable and that no DNA sequence variation was detected by randomly amplified polymorphic DNA (RAPD) assay after cryopreservation. In addition, the methylation sensitive amplified polymorphism (MSAP) assay indicated that cryopreservation caused a significant change in DNA methylation status. PMID- 11912506 TI - Analysis of ploidy and the patterns of amplified fragment length polymorphism and methylation sensitive amplified polymorphism in strawberry plants recovered from cryopreservation. AB - Shoot-tips of 10 strawberry genotypes were successfully cryopreserved using a modified encapsulation-dehydration method. All genotypes survived cryopreservation with high survival and regeneration rates. Eight Joho single-bud sibling lines were established as a model system for genetic analysis. Although cytological examination found chromosomal variation in both non-cryopreserved and cryopreserved samples, the ploidy constitution remained relatively stable after cryopreservation. DNA samples digested with MseI and PstI were used for amplified fragmentation length polymorphism (AFLP) assay. In 16 primer combinations, only one, namely, PCCA-MCAG, detected one site where band pattern changed after cryopreservation, which might be contributed to the change in DNA methylation status at PstI recognition site. Methylation sensitive amplified polymorphism (MSAP) assay was carried out for further investigation on the influence of cryopreservation on DNA methylation status. It was found that cryopreservation induced a significant change in DNA methylation status. PMID- 11912507 TI - Cryopreservation and xenotransplantation studies of microencapsulated rat pancreatic islets. AB - Islets of Langerhans were isolated from the Sprague Dawley rat pancreas digested by injected collagenase, and purified by Ficoll density gradient centrifugation. In order to make smaller and more uniform microencapsulated islets, we designed a special high-voltage electrostatic microcapsule generator. The effects of operational parameters of the generator on the size and the uniformity of microcapsules were analyzed, such as the voltage, the plunger speed of suspension delivery to the needle tip, the distance between needle tip and solution surface. The optimal parameter combinations for making microcapsules are: 5 kV of voltage, 50 mm/h of the plunger speed, and 20 mm distance. The high-voltage electric system can produce uniform microcapsules with diameters ranging from 0.3 to approximately 0.5 mm, which are smaller and more uniform than those produced by air-jet system. A comparison of the cryopreservation effects between microencapsulated islets and unencapsulated islets showed that the microcapsules can protect the fragile islets from freezing damage, and increase the retrieval rate from 68.5% to 92.6%. Xenotransplantation of the cryopreserved rat islets resulted in the normalization of the metabolic blood glucose of the diabetic mice for 90 days, whereas the unencapsulated islets were easily fragmented and lost during the freezing process. They only reversed hyperglycemia for less than 3-5 days. PMID- 11912508 TI - Ammonium nitrate in the culture medium influences regeneration potential of cryopreserved shoot tips of Holostemma annulare. AB - Influence of NH4NO3 in the pre-freeze and post-freeze culture medium and 2 or 30 day preconditioning in the presence of 0.5 M sucrose on regeneration of shoot tips of Holostemma annulare following cryopreservation using an encapsulation dehydration protocol was studied. A long preconditioning phase of 30 days significantly reduced tissue water and improved post-freeze recovery of shoot tips. Under the long preconditioning treatment, Murashige & Skoog (MS) medium free of NH4NO3 (MS-3) allowed maximum regeneration (59%) of liquid nitrogen (LN) exposed shoot tips with less frequency of callusing (10.4%) after 45 days of post freeze culture. Corresponding desiccated control shoot tips showed 85-90% regeneration. A 3.75 mM NH4NO3 concentration (MS-4) favoured 72-89% and 43-47% regeneration after desiccation and LN exposure respectively. The standard MS medium with 20.6 mM NH4NO3 (MS-1) allowed poor regeneration after desiccation (39 53%) as well as LN exposure (8-23%). The study reveals the importance of reducing ammonium nitrate in the culture medium to get maximum recovery of cryopreserved shoot tips of Holostemma annulare. PMID- 11912509 TI - Cryopreservation of cultivated and wild Arachis species embryonic axes using desiccation and vitrification methods. AB - The effects of two methods of cryopreservation involving chemical vitrification and air desiccation) were studied on isolated embryonic axes of A. hypogaea. Vitrification with PVS2 and desiccation in a laminar flow cabinet resulted in high levels (70-90%) of whole plant recovery after cryopreservation. A desiccation protocol based on 1h exposure of explants to the air flow was successfully applied to six wild species of section Extranervosae, resulting in recovery levels of 70-90% after liquid nitrogen treatment. PMID- 11912511 TI - Nitrergic innervation of the normal gut and in motility disorders of childhood. PMID- 11912510 TI - Missense mutations in GJB2 encoding connexin-26 cause the ectodermal dysplasia keratitis-ichthyosis-deafness syndrome. AB - Keratitis-ichthyosis-deafness syndrome (KID) is a rare ectodermal dysplasia characterized by vascularizing keratitis, profound sensorineural hearing loss (SNHL), and progressive erythrokeratoderma, a clinical triad that indicates a failure in development and differentiation of multiple stratifying epithelia. Here, we provide compelling evidence that KID is caused by heterozygous missense mutations in the connexin-26 gene, GJB2. In each of 10 patients with KID, we identified a point mutation leading to substitution of conserved residues in the cytoplasmic amino terminus or first extracellular domain of Cx26. One of these mutations was detected in six unrelated sporadic case subjects and also segregated in one family with vertical transmission of KID. These results indicate the presence of a common, recurrent mutation and establish its autosomal dominant nature. Cx26 and the closely related Cx30 showed differential expression in epidermal, adnexal, and corneal epithelia but were not significantly altered in lesional skin. However, mutant Cx26 was incapable of inducing intercellular coupling in vitro, which indicates its functional impairment. Our data reveal striking genotype-phenotype correlations and demonstrate that dominant GJB2 mutations can disturb the gap junction system of one or several ectodermal epithelia, thereby producing multiple phenotypes: nonsyndromic SNHL, syndromic SNHL with palmoplantar keratoderma, and KID. Decreased host defense and increased carcinogenic potential in KID illustrate that gap junction communication plays not only a crucial role in epithelial homeostasis and differentiation but also in immune response and epidermal carcinogenesis. PMID- 11912512 TI - Management of splenic trauma in the pediatric hemophiliac patient: Case series and review of the literature. AB - In July and August 1998, 3 patients who attend the Hemophilia Treatment Center required emergency admission to the authors' hospital for management of hemorrhagic shock caused by splenic injury. Computed tomography was used to diagnose and grade the splenic injuries, which ranged from II to IV on the organ injury scale. Two patients had Christmas disease (Factor IX deficiency) and were treated with splenorrhaphy and factor IX replacement. One patient who has severe von Willebrand disease (Type 3) had grade II splenic injury that required splenectomy to secure hemostasis. The coagulopathic deficiency was aggressively treated in each patient. All patients required operative intervention with attempted splenorrhaphy. All patients survived their operative experience, and none suffered a rebleeding episode. With correction of the coagulopathy throughout the perioperative period and local hemostatic control by operative techniques, salvage procedures for splenic injury were successful for 2 of these 3 patients. PMID- 11912513 TI - Falling televisions: The hidden danger for children. AB - BACKGROUND/PURPOSE: The United States Consumer Product Safety Commission (USCPSC) recently has reported a significant number of injuries and deaths in the home related to televisions (TV) falling on children. To date, little is known regarding the significance of this mechanism of injury in childhood trauma. The current investigation was designed to examine the risk factors, spectrum of injuries, and operative intervention required in children injured by falling televisions. METHODS: The records of all patients 0 to 16 years of age with television-related injuries and entered in the Pennsylvania Trauma Outcome Study (PTOS) between 1989 and 1999 were reviewed. The authors examined Glascow coma scale (GCS), injury severity score (ISS), length of hospital stay (LOS), major injuries sustained, and operative procedures performed. Fourteen of the children in the PTOS were seen at the Benedum Pediatric Trauma Center at the Children's Hospital of Pittsburgh. In these 14 children, a review of the medical records was performed for a detailed description of the accident scenario. RESULTS: Forty three children sustained television-related injuries during this period. Nearly 56% of these children were .1). Data indicate that metformin decreases the serum leptin concentration even without affecting body weight and body composition in normal-weight men. PMID- 11912568 TI - The evolving role of topiramate among other mood stabilizers in the management of bipolar disorder. AB - OBJECTIVES: Topiramate, a structurally novel anticonvulsant, is being evaluated for other neurological conditions such as migraine, neuropathic pain, and essential tremor, and also for psychiatric conditions such as bipolar disorder, bulimia, post-traumatic stress disorder, and schizoaffective disorder, in addition to obesity. This article will focus on the use of topiramate for bipolar disorder. METHODS: The pharmacological profile of topiramate is compared to other established and putative mood stabilizers, and a rationale for its use in bipolar disorder is presented. Data from open clinical trials of topiramate for depression, mania, and rapid-cycling bipolar disorder are summarized. Preliminary data from one pilot dose-finding, double-blind, random-assignment, placebo controlled, 3-week parallel group study of two doses of topiramate for acute bipolar I mania is reported. Safety data regarding topiramate was reviewed. Finally, the potential place of this agent in bipolar illness is considered. RESULTS: The pharmacological advantages for topiramate are low protein binding, minimal hepatic metabolism and mainly unchanged renal excretion, a 24-h half life, and minimal drug interactions. Open clinical studies suggest a 50-65% response for refractory bipolar mania, and a 40-56% response for refractory bipolar depression in mainly add-on treatment. Open clinical studies of topiramate for rapid-cycling subjects and those for comorbid bulimia, substance abuse, post-traumatic stress, migraine, and obesity report effectiveness. The primary efficacy endpoint data (change from baseline Y-MRS total scores) of the placebo-controlled, random assignment parallel group phase II dose-finding study were not statistically significant. However, once the antidepressant-associated manias (28 of the sample, of 97 subjects) were excluded from the controlled study, the post-hoc analyses indicated the higher dose (512 mg/day) topiramate treatment group showed a statistically significant reduction in endpoint Y-MRS change scores as compared to placebo (p < 0.03). Adverse effects of topiramate in bipolar subjects include attention, concentration and memory problems, fatigue, sedation, transient paraesthesias, nausea, and anorexia. Some subjects experience word-finding difficulty. Weight loss may be seen in several topiramate-treated subjects with bipolar disorder. CONCLUSIONS: Topiramate appears to show promise as an addition to the agents available to treat bipolar disorder. More definitive controlled data on the efficacy of topiramate in the acute and continuation phases as well as for the prophylaxis either as monotherapy or as combination treatment of bipolar disorder are ongoing, and the results are awaited. PMID- 11912569 TI - Rate of switch in bipolar patients prospectively treated with second-generation antidepressants as augmentation to mood stabilizers. AB - INTRODUCTION: Bipolar patients with breakthrough major depressive episodes despite ongoing adequately-dosed mood stabilizer medication were randomized in a double-blind manner to one of three antidepressants with different mechanisms of action: bupropion, sertraline, or venlafaxine. Preliminary data are presented on the switch rates into hypomania or mania for the antidepressants as a group prior to unblinding the specific individual drug efficacy and tolerability data in this ongoing clinical trial. METHODS: Subjects included 64 bipolar patients who participated at five sites in a 10-week double-blind trial for depression and a 1 year blinded continuation maintenance phase for responders. Nonresponders were re randomized such that there were 95 acute treatment phases. In the acute phase, doses were titrated to clinical response, side effects, or maximum dose of bupropion (450 mg/day), sertraline (200 mg/day), or venlafaxine (375 mg/day). Daily ratings on the National Institute of Mental Health-Life Chart Methodology (NIMH-LCM) were inspected for the degree of improvement on the Clinical Global Impressions scale as revised for bipolar illness (CGI-BP) and the occurrence of hypomania or mania. RESULTS: Thirty-five (37%) of the 95 acute treatment phases were associated with a much or very much improved rating in depression on the CGI BP. Thirteen (14%) of these 95 acute trials of antidepressants as adjuncts to mood stabilizers were associated with switches, seven into hypomania and six into mania. Forty-two patients elected to go into the continuation phase in 48 instances. Sixteen (33%) of the continuation phase trials were associated with mood switches, 10 into hypomania and six into mania. CONCLUSIONS: In this randomized double-blind prospective study of three second-generation antidepressants (bupropion, sertraline, and venlafaxine) in bipolar patients whose depression broke through ongoing treatment with mood stabilizers, switches into hypomania or mania occurred in 14% of the acute phases and 33% of the continuation phases. Individual data on each drug will be assessed in the next phase of the study after more subjects are recruited and the blind is broken. PMID- 11912570 TI - Beating the Elijah syndrome. PMID- 11912571 TI - Spiritual care. Assessment & intervention. PMID- 11912572 TI - Faith in God. Help for partners in pain. PMID- 11912573 TI - Miracles of mercy. PMID- 11912574 TI - Nursing & ministry. Complementary care. PMID- 11912575 TI - Mentoring. Sharing our wisdom. PMID- 11912577 TI - Jesus, our mentoring model. PMID- 11912578 TI - Teen treasures. Mentoring in action. PMID- 11912579 TI - Camp nurse. PMID- 11912580 TI - Sunday shift. What am I doing here? PMID- 11912581 TI - Code 99 & the cross. PMID- 11912582 TI - Work as worship. PMID- 11912583 TI - CPCs caring for the desperate. PMID- 11912584 TI - Who is my neighbor? Reflections on serving the homeless. PMID- 11912585 TI - Creative clinical in the inner city. PMID- 11912586 TI - Man down! A nurse goes to jail. PMID- 11912587 TI - Instruments of God's peace. PMID- 11912588 TI - Living simply. Choosing to become poor. PMID- 11912590 TI - Imprisoned. Why can't you see me? PMID- 11912589 TI - Dinghy what? Confronting the consequences of caring. PMID- 11912592 TI - Code four & the ministry of a marginalized man. PMID- 11912593 TI - Compassion in crisis. Aftermath of the Nairobi bombing. PMID- 11912594 TI - To be a nurse. From foolishness to faith. PMID- 11912596 TI - Renewed to walk with the poor. PMID- 11912597 TI - Packing for the journey. Character traits for transcultural care. AB - Eight-four pounds," the seaplane attendant announced approvingly. As two nursing students and an instructor, we were on our way to a remote First Nations village in northern Canada as part of a transcultural clinical course at a Canadian Christian university. Our groceries for three weeks were boxed up and weighed to ensure that the carrying capacity of the seaplane was not exceeded. We had packed carefully, giving priority to items that were nutritious, lightweight and compact. We hoped we had the essentials. We felt prepared. PMID- 11912599 TI - On the team. PMID- 11912598 TI - Crosscultural clinical. PMID- 11912600 TI - Tree of hope. PMID- 11912601 TI - Set free from fear. PMID- 11912602 TI - When God took the reservation by storm. PMID- 11912603 TI - Medicine men. PMID- 11912604 TI - Have profession, will travel. PMID- 11912606 TI - God at work. Attitude overhaul in Russia. PMID- 11912607 TI - Finding grace in despair. PMID- 11912608 TI - Having nothing, possessing everything. PMID- 11912609 TI - Embracing a people: the joy of incarnational ministry. PMID- 11912610 TI - Drink of life: oral rehydration therapy. PMID- 11912611 TI - Dissemination of diabetes care guidelines: lessons learned from community health centers. AB - PURPOSE: This study was conducted to evaluate the impact of a provider problem based learning (PBL) intervention on screening for complications of diabetes in community health centers. METHODS: A successive sampling design was used to compare selected standards of diabetes care delivered preintervention with the care delivered postintervention at 2 community health centers and 1 comparison centers. Two randomly assigned intervention sites received a PBL intervention focused on care guidelines for prevention of diabetes complications, with telephone follow-up over 12 months. Effects of the intervention were determined from an audit of 200 charts from each site. RESULTS: The odds of having a glycosylated hemoglobin test more than doubled from preintervention to postintervention, and the odds of having a foot examination more than tripled across centers. Measurement of creatinine and glycosylated hemoglobin were associated; the odds of having one test tripled when the other had been measured. Rates for documentation of patient education were significantly lower at the intervention site where free patient education booklets were distributed. CONCLUSIONS: Improvements in diabetes care were not consistent among community health centers. Interventions involving system and policy changes may be more effective in implementing and sustaining improvements than just provider education. PMID- 11912613 TI - Values central to advertising in The Diabetes Educator. PMID- 11912612 TI - Physician assistant students and diabetes: evaluation of attitudes and beliefs. AB - PURPOSE: Physician assistants are assuming a greater role in patient care in the US health system. The objective of this study was to examine attitudes and beliefs about diabetes among physician assistant trainees. METHODS: A survey of 3 currently enrolled classes of physician assistant students was conducted using the Diabetes Attitude Survey (DAS, version 3). An additional question was presented to gather information about the level of hyperglycemia at which students would intensify diabetes therapy. RESULTS: On average, students scored high on all subscales, indicating general agreement with the attitudes examined by the DAS. For 3 subscales (seriousness of type 2 diabetes, value of tight glucose control, and patient autonomy), significant differences were seen across year of training. When asked about the level of glucose control at which they would advance therapy, a wide range of responses occurred, with some being out of target. CONCLUSIONS: Physician assistant students had favorable attitudes regarding type 2 diabetes. However, deficits appear to exist in understanding when to advance therapy. More studies on physician assistant students' knowledge of diabetes standards of care are required. PMID- 11912614 TI - Diabetes mellitus and cardiovascular disease. PMID- 11912615 TI - Community partnerships for parent-to-parent support. PMID- 11912617 TI - Gestational diabetes mellitus: diagnosis, treatment, and beyond. PMID- 11912616 TI - Diabetes Education Programs for African American Women: what works? PMID- 11912618 TI - Managing type 1 diabetes during pregnancy. PMID- 11912619 TI - Development of a theory-based daily activity intervention for individuals with type 2 diabetes. AB - PURPOSE: This article describes a theory-driven approach to developing a physical activity intervention for sedentary individuals with type 2 diabetes. METHODS: Development of the intervention was based on 6 essential elements of program theory: problem definition, critical inputs, mediating processes, expected outcomes, extraneous factors, and implementation issues. Each element was formulated based on available literature and in collaboration with both intended service deliverers (diabetes educators) and recipients (sedentary persons with type 2 diabetes). RESULTS: Diabetes education requires a simple physical activity intervention template that is feasible, acceptable, and effective in a variety of settings. Successful programs are individualized, specific, flexible, and based on walking. Pedometers have potential as self-monitoring and feedback tools. The primary expected outcome is an increase in physical activity, specifically walking. Behavior modification and social support are critical to adoption and adherence. CONCLUSIONS: Theory-driven interventions specify what works for whom and under what conditions of delivery. The underlying theory guides the evaluation, refinement, and clinical replication of an intervention. Recruitment, delivery, and follow-up are real-world implementation issues. PMID- 11912621 TI - [Uprooting measles from Finland]. PMID- 11912620 TI - A community diabetes educator course for the unemployed in South Auckland, New Zealand. AB - PURPOSE: This paper describes an education program that targeted long-term unemployed people from the community and trained them to work as diabetes educators in their own communities in an attempt to address issues of cultural appropriateness. METHODS: Government funding was obtained to conduct two 22-week training courses for people who had been selected by their communities. These courses built on participants' existing cultural skills and provided appropriate diabetes training. RESULTS: The results indicate that the courses were successful both in creating a cadre of culturally acceptable diabetes educators and providing employment for course participants. CONCLUSIONS: Previously unemployed lay people are able to provide diabetes education in the primary prevention and group settings. Such individuals are able to incorporate extensive cultural skills in their work. PMID- 11912622 TI - [General anesthesia in pediatric dentistry]. PMID- 11912623 TI - [What happens to the organs in space?]. PMID- 11912624 TI - [How to auscultate the lungs in a pediatric patient?]. PMID- 11912625 TI - [The role of angiotensin II receptors in the pathogenesis of cardiovascular diseases]. PMID- 11912626 TI - [Diagnosing pulmonary embolism]. PMID- 11912627 TI - [Breast cancer diagnostics. Finnish Breast Cancer Group]. PMID- 11912628 TI - [Intrauterine infections and the fetus]. PMID- 11912629 TI - [The second basket]. PMID- 11912630 TI - [Spontaneous intracranial hypotension--a treatable cause of headache]. PMID- 11912631 TI - [Pain in the fetus--true or overemphatetic imagination?]. PMID- 11912632 TI - [Malaria death in Finland]. PMID- 11912633 TI - [Views about the surgical treatment of breast cancer and its renewal]. PMID- 11912634 TI - [Was there mycobacterial infection behind the bone necrosis?]. PMID- 11912635 TI - [The safety of sildenafil]. PMID- 11912636 TI - [Annals of Medicine speedily to international recognition]. PMID- 11912637 TI - [Treatment of prostate cancer. Practice recommendations from the Finnish Urological Association]. PMID- 11912638 TI - [Forensic medicine yesterday--forensic sciences tomorrow]. PMID- 11912639 TI - [Did the O.J. Simpson case teach us anything--when science meets the courtroom]. PMID- 11912640 TI - [Why the causes of death have to be investigated?]. PMID- 11912641 TI - [Autopsies as a guarantee of legal protection and quality of patient care]. PMID- 11912642 TI - [Drugs, poisons, illicit drugs or alcohol again?]. PMID- 11912643 TI - [A living human being and forensic medical investigations]. PMID- 11912644 TI - [What can the teeth tell?]. PMID- 11912645 TI - [Molecular genetics revolutionized forensic medicine]. PMID- 11912646 TI - [Laboratory helps the crime investigators]. PMID- 11912647 TI - Congregational care. Reaching out to the elderly. PMID- 11912648 TI - Prayer research. What are we measuring? PMID- 11912649 TI - Dancing with Dianna. PMID- 11912650 TI - When hope comes hard. PMID- 11912651 TI - Nursing the soul. A team approach. PMID- 11912652 TI - Does believing make us healthy? PMID- 11912653 TI - Give me children or I'll die. Easing the spiritual pain of infertility. PMID- 11912655 TI - My name is Jim. Do you know me? PMID- 11912654 TI - Formula for a miracle. PMID- 11912656 TI - Where was Jesus tonight? PMID- 11912657 TI - The healing power of prayer. PMID- 11912658 TI - Breaking the spiritual care barrier. PMID- 11912659 TI - Off-label use of prescription drugs. PMID- 11912660 TI - How I spent my summer vacation. PMID- 11912661 TI - Camp Discovery: the first 7 years. PMID- 11912662 TI - Camp happiness. AB - In the outside world, children with serious skin conditions often face isolation and loneliness. Their peers give them nicknames, uninformed adults ask difficult questions. Once they get to Camp Horizon, however, they forget their disease, and swim, boat, stargaze, sing around the fire. They meet other children just like themselves. According to one boy, 'This place is like Heaven on Earth for me." PMID- 11912663 TI - What's your assessment? AB - The "What's Your Assessment?" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11912664 TI - Systemic glucocorticosteroid therapy in dermatology. AB - Glucocorticosteroids (GCS) have been frequently used for a number of dermatologic conditions since the early 1950s. These drugs continue to be very efficacious, yet there are serious side effects associated with their long-term use. Significant complications produced by GCS can be minimized by proper management and precautions. All health care professionals should be aware of these strategies and nursing personnel can play a vital role in helping patients adhere to the guidelines of preventing adverse events. PMID- 11912665 TI - Cosmetic rehabilitation. AB - Appearance is one of the most powerful factors influencing social interactions with others. What patients with disfigurements experience from society on a daily basis is a very real stigma. Cosmetic rehabilitation is a system of cosmetic techniques devised for patients to use to assist themselves to cope constructively with the psychological and physical trauma of their disfigurements. PMID- 11912666 TI - New hormonal therapies treat persistent acne in women. PMID- 11912667 TI - Common skin conditions may indicate HIV infection in women. PMID- 11912668 TI - Rosacea now successfully controlled with therapy and lifestyle changes. PMID- 11912669 TI - The leader as a retention specialist. AB - Retention problems will plague us for the near future. Blanchard and Waghorn (1997) believe that everyone wants to be magnificent, and not just ordinary. Unfortunately, often we expect only the ordinary and not the magnificient. People respond to our expectations. We can help staff to achieve magnificence by providing unconditional love and remembering that people are magnificent. Sometimes their behavior is a problem and certain things block the expression of their greatness, but all people are magnificent. If we create "sanctuaries of caring" where nurses and others are given the opportunity to achieve their innate nobility and magnificence, people will flock to participate in the noble and magnificent experience of patient care. PMID- 11912670 TI - An experimental disconnection approach to a function of consciousness. PMID- 11912671 TI - Experimental models of partial lesion of rat spinal cord to investigate neurodegeneration, glial activation, and behavior impairments. AB - The article demonstrates two experimental models of spinal cord partial injury in rats: a contuse model promoted by the NYU impactor system and a partial hemitransection model achieved by a stereotaxic-positioned adjustable wire knife. By means of a defined impact weight (10 g) and a digital optical potentiometer linked to a computer, the impactor transferred and registered a moderate or a severe contusion to the rat spinal cord at a low thoracic level after dropping the weight from distances of 25 mm and 50 mm, respectively, to the dorsal surface of the exposed dura spinal cord. Impact curve was calculated and the parameters of the trauma, like impact velocity, cord compression distance and cord compression rates were obtained in order to demonstrate trauma severity. To promote partial hemitransection, rats were positioned in a spinal cord unit of a stereotaxic apparatus and lesion was made with the adjustable wire knife spatially oriented. By means of a computerized infrared motion sensor-home cage activity monitor and a noncomputerized evaluation of motor behavior using the inclined plane and the motor score of Tarlov tests, behavior was analyzed in an acute period postlesion. Rats were sacrificed and spinal cords were processed for routine staining to show neurons and for GFAP and OX42 immunohistochemistry to demonstrate glial cells. The tissue labelings were quantified using computer assisted stereology by means of an optical disector and microdensitometric image analysis by means of quantification of gray values of discriminated profiles. While partial hemitransection model favored a more accurate control of the lesion location, the contuse model allowed us to perform different degrees of lesion severity. A close correlation between behavioral impairment and severity of trauma was seen in the rats submitted to spinal cord contusion. The stereologic lesion index showed a correlation between severity of trauma and tissue damage by 7 days and demonstrated a time-dependent secondary degeneration after moderate but not after severe spinal cord contusion from 7 to 30 days after injury. Long lasting activations of astrocytes and microglia seen by persisted increases in the specific mean gray values of immunoreactivities were also found in all levels of the white and gray matters of the partial hemitransected spinal cord until 3 months postinjury which can be related to wound/repair events and paracrine trophic support to spinal cord remaining neurons. The results showed that controlled partial lesions may provide an important toll to study trophism and plasticity in the spinal cord. PMID- 11912672 TI - A visual learning and memory test for preoperative evaluation of patients with temporal lobe epilepsy. AB - We used a modified version of the visual DCS (mDCS) test to study patterns of learning, free recall, and recognition capacity in patients with temporal lobe epilepsy (TLE) (N = 24) and controls (N = 38). The patients were under consideration for epilepsy surgery. The mDCS differentiated left from right TLE patients in all aspects of visuospatial learning and delayed recall. The mDCS revealed lower performance for TLE patients as compared with controls. The present results indicate that the mDCS may be a preoperative tool to differentiate left from right TLE. PMID- 11912673 TI - Effect of time scales on the unfolding of neural attractors. AB - A study of the effect of time scales in brain dynamics on the unfolding of the attractors in the phase space, reconstructed by a time delay embedding of the EEG signal, was carried out. Applying the techniques of nonlinear time series analysis, the unfolding rate of the system attractor was determined by analyzing the variation of the correlation dimension parameter and subjecting it to a bi parametric fit. The behavior of the parameter, which measures the rate of unfolding, was monitored for varying time scales in two cases: (a) normal eyes closed condition and (b) the pathological case of epilepsy. Significant results were obtained. PMID- 11912674 TI - Differences between right- and left-femoral bone mineral densities in right- and left-handed men and women. AB - To test whether handedness has an asymmetric effect on femur bone mineral density (BMD), the right- and left-proximal femur BMDs were compared in 124 right- and 23 left-handed university students. In the right-handed men, the mean left-femur BMD was significantly greater than the right-femur. In the left-handed men, the mean right-femur BMD was significantly greater than the left-femur. There were no significant differences between BMDs from the right- and left-femurs in women. These results suggest that hand preference may be related to asymmetry in BMD, but only for men. PMID- 11912675 TI - Integration of neuropsychological and cognitive theory in rehabilitation. AB - Clinical neuropsychology has focused on the diagnosis and localization of brain damage. Much of the early progress in the field relied upon an atheoretical actuarial approach. Recently a link between neuropsychology and cognitive psychology has emerged. From the early focus upon brain-behavior relationships, neurocognitive theories have emerged to explain observed brain functions. Although many researchers have enjoyed the collaboration between neuropsychology and cognitive psychology, others continue to investigate functions without considering the merging of finds. This article reviews integrative elements of cognitive and neuropsychological research and the emergence of cognitive neuropsychology. PMID- 11912676 TI - Report of normative sensory and motor performance in children using a standardized battery. AB - This study provided normative data for sensory and motor functioning of children from 2-15 years of age. Two hundred eighty-eight children without a history of neurological or psychiatric evaluation, diagnosis, or treatment served as subjects. Each child was administered the Dean-Woodcock Sensory and Motor Battery. This measure consists of 18 individual subtests. Nine subtests reflect visual, auditory, or tactile perception. The remaining 9 subtests involve motor functioning, with 3 of the subtests devoted to subcortical functioning. The results showed that the most rapid development occurred between the ages of 4 and 7 years. Indeed, some 76% of the sensory-motor tasks administered reached a plateau by age 7. PMID- 11912677 TI - A confirmatory factor analysis of the cognitive capacity screening examination in a clinical sample. AB - Structured mental status examinations offer several advantages over unstructured mental status examinations; however, few have been subjected to advanced psychometric analysis. Confirmatory factor analysis empirically tests the predictive validity of individual test indices within an a priori methodological framework. With such analysis, one can test hypotheses about the structure of latent variability within a given data set. The purpose of this study was to perform a confirmatory factor analysis of the Cognitive Capacity Screening Examination, the most comprehensive of the brief structured mental status examinations. A confirmatory factor analysis of the Cognitive Capacity Screening Examination (CCSE) was performed by applying LISREL 7 to a sample of 924 male veterans, 409 patients from a chemical dependence treatment program, and 515 individuals from a psychology consultation service. Constructs were derived from previous exploratory analysis of the scale. The results of the confirmatory factor analysis and three indices of model fit support an 11 factor model more complex than that originally formulated for the CCSE. However, three of these factors (digit span with interference, complex mental mathematics, and verbal memory) were more sensitive to impairment than other factors, accounting for over 90% of the CCSE total score variance. Although the CCSE is a more complex test than originally envisioned by its designers, it may not be necessary to give all items on the test. Either a subset of the CCSE items (the CCSE-A) or a relatively brief, informal mental status exam may be adequate for many patients. PMID- 11912678 TI - Correlation of the LNNB-III with the WAIS-III in a mixed psychiatric and brain injured population. AB - The purpose of the study was to examine the relationship between the Luria Nebraska Neuropsychological Battery-Third Edition (LNNB-III) and the Wechsler Adult Intelligence Scale-Third Edition (WAIS-III). Participants were 85 adults referred for neuropsychological evaluation. The mean age of participants was 38.73 years (SD = 16.54) and average education was 13.07 years (SD = 2.60). The sample was predominantly female (52.9%), right-handed (86.3%), and Caucasian (68.6%), with the remainder of the population classified as Hispanic (13.7%), African-American (5.9%), or other (11.8%). Diagnoses included 26% psychiatric disorders, 64% neurological disorders, and 10% with no diagnosis. Pearson product correlation yielded a number of significant relationships between the WAIS-III IQ scores and the LNNB-III scales. The highest correlations were with the LNNB Intelligence, Visual-Spatial, Complex Auditory, and Arithmetic scales. Additionally, significant correlations were found between the WAIS-III subtests and a moderate proportion of the LNNB-III subtests. Correlations were also reported for the new WAIS-III scales, Letter-Number Sequencing and Matrix Reasoning. The results suggest that similar abilities are being assessed on both tests. These findings allow clinicians to not only evaluate the consistency of performance across this testing battery, but provide a useful screening instrument for intelligence. PMID- 11912679 TI - Assessment of cognitive functioning in men who batter. AB - The present investigation examined neuropsychological functioning in 50 male batterers court-ordered into treatment and 23 nonpatient controls. Subjects were administered a neuropsychological screening battery consisting of the Screening Test for the Luria-Nebraska Neuropsychological Battery, the Stroop Color and Word Test, two memory subtests from the Luria-Nebraska Neuropsychological Battery-III (Figural Memory and Delayed Figural Memory), and two subtests from the Halstead Reitan Neuropsychological Battery (Trails A & B). Subjects were categorized as having neuropsychological dysfunction if their scores exceeded the statistical cut offs on two or more subtests. Results indicated that 24 (48%) of the male batterers exhibited cognitive dysfunction, as compared to only 1 (4.3%) of the nonpatient controls. Inspection of individual neuropsychological measures indicated poorer performance across all subtests for impaired male batterers as compared to both nonimpaired batterers and normal controls. In contrast, no significant differences on any of these measures emerged between nonimpaired male batterers and normal controls. Implications for the appropriate screening and treatment of male batterers are discussed. PMID- 11912680 TI - Effect of trichloroethylene on spatiotemporal pattern of LTP in mouse hippocampal slices. AB - The effect of trichloroethylene (TCE) on long-term potentiation (LTP) was studied using both electrical and optical recording. The hippocampi from mice injected with 300 mg/kg or 1000 mg/kg TCE were sliced 24 h after administration. The field potential from the CAI was recorded. After the application of tetanus, population spikes (PS) were potentiated in all groups, but the post-per-pre ratio of PS was smaller in TCE groups than in the control. Optical recording was also carried out in 1000 mg/kg TCE-injected mice and a new analytical method using a high speed camera was employed. After the induction of tetanus, the optical signal was potentiated in both TCE and control groups. However, the post-per-pre ratio of the optical signals and response area were smaller in the TCE groups than in the control. It was suggested that the impairment of LTP is one of the mechanisms of the impairment of immediate memory after acute exposure to TCE in humans. PMID- 11912681 TI - [Pancreatic excisions for chronic pancreatitis and cancer: their rationale for "factual" surgery. Evidence-based medicine]. AB - Despite significant improvement in the results of pancreatoduodenecomy over recent years, the Whipple procedure and its main modifications still has a poor reputation. Based on the principles of evidence-based medicine, we reviewed the current status of pancreatoduodenectomy for pancreatic cancer and chronic pancreatitis. Mortality of pancreatoduodenectomy has declined to less than 5% for chronic pancreatitis and to 3-5% for pancreatic cancer. In contrast, overall morbidity remains high, ranging from 20% to 70%. Delayed gastric emptying accounts for almost 50% of all complications. Major relief of pain is achieved in 70% to 100% of patients with chronic pancreatitis. Overall 5-year survival for patients with pancreatic cancer remains poor, ranging from 5% to 15%, with a median survival of 13 to 17 months. Mortality ad morbidity are not related to the type of pancreatoduodenectomy, however patients with pancreatic cancer tend to have a higher risk for complications. Extended lymph node dissection and portal vein resection can be performed with similar mortality and morbidity compared with standard procedures, however without any survival benefit in the long-term course. PMID- 11912682 TI - [Fortuitous discovery of gallbladder cancer]. AB - The prognosis of gallbladder cancer is basically dependent on the histological stage at diagnosis. In practice, the discovery of a small cancer of the bladder, generally during cholecystectomy give the patient a better care for curative treatment. The advent of laparoscopy has increased the number of cholecstectomies and could increase the frequency of this situation but also raises the difficult problem of metastatic dissemination. In the literature the figures on parietal metastasis after laparoscopy have ranged from 125% to 19%. The median delay to diagnosis of recurrence is 6 months. The cause of this phenomenon (role of the pneumoperitoneum) remains poorly elucidated. Risk factors for the development of a metastasis on the trocar orifice are: rupture of the gallbladder perioperatively and extraction of the gallbladder without protection. It is important to keep in mind this exceptional but serious risk and apply rigorous operative technique. In case of suspected gallbladder we do not advocate laparoscopy. Surgery (hepatectomy, lymphodenectomy, possibly resection of the biliary tract) would be indicted for all stages except pTis and T1a, taking into consideration the localization of the tumor and the patient's general status. It is also classical to recommend resection of the trocar orifices after laparoscopic cholecystectomy. There is a dual challenge today for small-sized gallbladder cancer: improving treatment and avoiding poorer prognosis due to the specific problems raised by laparoscopy. PMID- 11912684 TI - [Diagnostic indications of upper gastrointestinal endoscopy in esophago-gastro duodenal disease in adults. Excluding echoendoscopy and enteroscopy]. PMID- 11912683 TI - [Transplantation of isolated hepatocytes, is it an alternative for total liver transplantation? On the treatment of hereditary hepatic metabolic diseases]. AB - Since 1976, it has been demonstrated in the small animal that isolated hepatocytes transferred into the liver can integrate into the recipient organ, survive and perform differentiated functions. This cell therapy technique can be used to partially correct for hereditary metabolic disorders in animal models of human hereditary metabolic diseases. The first methods used for hepatocytes transplantation involved a mass of hepatocytes that was clearly to limited (about 0.5% and never more than 2%). Integration fo the hepathocytes was limited by poorly understood factors. The vary variable clinicial results in the eight trials performed in the 90s is a clear demonstration. Recent work has helped understand the mechanisms limiting the integration of transplanted hepatocytes in the recipient liver. This led to the concept of repopulation, increasing the percentage of the hepatocyte mass reconstituted during the transplantation. The goal is to give the transplanted hepatocytes a proliferation advantage over he native hepatocytes. Several models have been designed and are currently being evaluated for application in humans. We are not far from new clinical trials with hepatocyte transplantation in humans for the treatment hereditary metabolic diseases. For certain disease, 3 to 5% normally functioning hepatocytes in the recipient liver would be sufficient to achieve a clinically significant effect for the patient. PMID- 11912685 TI - [Right ileo-coloplasty after total esophagogastrectomy for caustic lesion]. PMID- 11912686 TI - [Laparoscopic promontofixation]. PMID- 11912687 TI - [Mechanical gastrointestinal suture. Part I: lineal sutures]. PMID- 11912688 TI - [Cholecystectomy: beware of the sliding of a sectorial spur]. PMID- 11912689 TI - [A 55-year-old woman seen after laparoscopic cholecystectomy]. PMID- 11912690 TI - [On the "diagnosis of appendiceal syndromes: for a rational management"]. PMID- 11912691 TI - Continuing education evaluation of behavior change. AB - Using the Cervero model of behavior change and questionnaires developed by Brigham, Ryan, and Elkins (1996), this study assessed the impact of a workshop on behavior change. Data on the individual professional (receptiveness), proposed change (feasibility), social system (climate), and program (quality), as well as self-report data about outcomes (expertise and actions) were included. Information on perceived expertise was obtained before the workshop, immediately after the workshop, and 3 months after the workshop. Information on actions initiated by the participants was obtained 3 months after the workshop. There were 70 participants in this study, with a variety of educational backgrounds and nursing experience. A matched test comparing expertise between time 1 and time 2, as well as time 1 and time 3 was significant at p = 0.00. There were significant correlations among the four variables influencing behavior change and actions and expertise. All except one participant in the study reported taking specific actions after the workshop. The mean number of actions taken was 4.0, with a range from 1 to 9. These findings indicate that a continuing education program can lead to behavior change. PMID- 11912692 TI - Level three evaluation of an education program designed to implement patient care redesign. AB - Patient care redesign is a process of reorienting healthcare institutions to a more patient-centered process. Inova Health System, a large Northern Virginia system, recently instituted such a redesign. The System's department of education was charged with designing a curriculum to introduce the new concept to all staff. This article analyzes the results of a survey that evaluated self-reported behaviors and attitudes of staff following attendance at the mandatory classes and discusses methodological aspects of the evaluation. The approach to evaluation discussed in this article provides a practical model for staff development educators when confronted with the task of evaluating results of educational programs. PMID- 11912693 TI - Total reorganization of a Nursing Policy and Procedure Manual. AB - This article describes the process of reorganizing an existing Nursing Policy and Procedure Manual. The process of restructuring the Nursing Policy and Procedure Committee and clarifying its priorities is also described. This discussion has significance for staff development educators because this type of project is often entrusted to them and few detailed guidelines exist for completing this task. PMID- 11912694 TI - Our dialogue: a case study in peer mentoring. AB - Two women from different parts of the organization are sitting together. They did not previously know each other and are from different professions and cultures. These women are interested in taking ownership of their own professional development. PMID- 11912695 TI - Microcomputer education for nursing: an approach to microcomputer education in a large tertiary care center. AB - Communication skills needed for the healthcare environment are changing. In the past, poor reading and writing skills made it difficult to use a paper chart. Current computer technology provides many new opportunities for communicating in the clinical setting. Computer literacy has become a necessary skill for efficient use of clinical computer systems. At this large tertiary care setting, a survey was developed and distributed to staff to assess learner needs. Nursing staff identified educational needs for basic computer knowledge, the ability to move between computer functions, and the skills to navigate through specific applications. Based on survey analysis, three educational classes were constructed that addressed the department's specific needs. PMID- 11912696 TI - From school to work: employer perceptions of nursing skills. AB - Questions periodically arise regarding clinical competencies and students' ability to perform in the employment setting after graduating from a baccalaureate nursing (BSN) program. This article describes a collaborative project between a nurse administrator in an acute care facility and a nurse educator in a baccalaureate nursing program. The project was designed to evaluate whether expected competencies of BSN graduates were evident in the work setting. Skills and behaviors of BSN graduates were evaluated by 68 nurses in an acute care setting. The findings provide data for nurse administrators to assist with planning staff development and continuing education programs for nursing staff. The results also provide data for nurse educators to assist with evaluation of effectiveness of nursing programs. PMID- 11912697 TI - Storytelling in staff development practice. PMID- 11912698 TI - Encouraging critical thinking. PMID- 11912699 TI - Finding strength to face breast cancer. Interview by Sandy Rensvold. PMID- 11912700 TI - Spiritual dehydration. Diagnosis & treatment. PMID- 11912701 TI - Does God heal today? PMID- 11912702 TI - "Who, me? Codependent?" Developing healthy relationships. PMID- 11912703 TI - I can't fix it! PMID- 11912704 TI - You are beautiful. PMID- 11912705 TI - Stop struggling. PMID- 11912706 TI - Footsteps on a rocky path. PMID- 11912707 TI - Emma & the med error. PMID- 11912708 TI - A strange premonition. PMID- 11912709 TI - When the going gets tough. PMID- 11912710 TI - Supporting the dying. The gift of an hour. PMID- 11912711 TI - Supporting the dying. Sacred memories. PMID- 11912712 TI - Supporting the dying. Last chance? PMID- 11912713 TI - Seeing God's plan unfold. PMID- 11912714 TI - Remembering Miss McBride. PMID- 11912715 TI - Front row seat on life. PMID- 11912716 TI - Lighting the lamp. PMID- 11912717 TI - Nurses Christian Fellowship. Building community. PMID- 11912718 TI - Three small steps toward racial reconciliation. PMID- 11912719 TI - Breast buddies. PMID- 11912720 TI - Disability in the body of Christ. PMID- 11912721 TI - A mission unlike any other. Life in a Kosovar refugee camp. PMID- 11912722 TI - Caring when you can't condone. The value of clinical role modeling. PMID- 11912723 TI - Enlarging our tents. PMID- 11912724 TI - Adolescent sexual crises. PMID- 11912725 TI - Empowering teens to say yes to abstinence. PMID- 11912726 TI - It's the livin' that's hard. PMID- 11912727 TI - The power of opposites. PMID- 11912728 TI - Discovering our common humanity. Developing partnerships in a Vietnamese community. PMID- 11912729 TI - Nita Barrow, RN. Called to lead a country. PMID- 11912730 TI - Flight nursing. A touch of hope. PMID- 11912731 TI - Family crisis. Lessons from the other side of the bed. PMID- 11912732 TI - Parish nursing. A call to integrity. PMID- 11912733 TI - Pragmatic partnership. PMID- 11912734 TI - Collaborating for mission. PMID- 11912735 TI - Parish nursing is ministry. PMID- 11912736 TI - Bearing another's burden. Five keys to therapeutic listening. PMID- 11912737 TI - Spiritual wellness in older women. PMID- 11912738 TI - A future with hope. PMID- 11912739 TI - Beyond band-AIDS. Empowering a Honduran community to care. PMID- 11912740 TI - Male nurse in a female field. Becoming colleagues. PMID- 11912741 TI - Captivated by God's call. Discovering the joy. PMID- 11912742 TI - Called to be consistently Christian. Verna Carson's personal story. PMID- 11912743 TI - [The elderly, health care resources and prioritization]. PMID- 11912744 TI - [For what is geriatrics needed?]. PMID- 11912745 TI - [The elderly, patient care and evidence-based medicine--overtreatment or undertreatment?]. PMID- 11912746 TI - [Not using the elephant's strength but your own personal strength]. PMID- 11912747 TI - [Why is it so difficult to diagnose illnesses in the elderly?]. PMID- 11912748 TI - [For how long is it possible and profitable to make the care of out-patients more effective?]. PMID- 11912749 TI - [When to institutionalized care?]. PMID- 11912750 TI - [The quality of long-term care can be measured, compared and improved]. PMID- 11912751 TI - [Efficacy of geriatric rehabilitation]. PMID- 11912752 TI - [A suspicious elderly person--even the paranoid ones have enemies]. PMID- 11912753 TI - [The depression of my old mother]. PMID- 11912754 TI - [Treatment alternatives of severe behavioral symptoms]. PMID- 11912755 TI - [When death is approaching]. PMID- 11912756 TI - [Life control in an ill elderly person]. PMID- 11912757 TI - [Who helps the elderly person--organizations and the elderly work]. PMID- 11912758 TI - [Back pain or back trouble?]. PMID- 11912759 TI - [Back pain--a significant public health problem]. PMID- 11912760 TI - [Does disturbed movement regulation cause chronic back trouble?]. PMID- 11912761 TI - [Organic background of the typical back pain]. PMID- 11912762 TI - [Psychic factors and back troubles]. PMID- 11912763 TI - [Risk factors and prevention of back aches]. PMID- 11912764 TI - [Physical examination of the patient with back pain at the physician's office]. PMID- 11912765 TI - [Diagnostic imaging in back pain]. PMID- 11912766 TI - [Conservative treatment of back pain]. PMID- 11912767 TI - [Surgical treatment of back pain is an exceptional solution--why does the number of back operations increase?]. PMID- 11912768 TI - [Surgical treatment of lower back intervertebral herniated disc]. PMID- 11912769 TI - [Prognosis of the patient with operated disc hernia]. PMID- 11912770 TI - [Lumbal spinal stenosis]. PMID- 11912771 TI - [Rheumatic back pains]. PMID- 11912772 TI - [Pregnancy and the back]. PMID- 11912773 TI - [Frequency and back ground of back pains in young persons]. PMID- 11912774 TI - [Investigation and therapy of spinal diseases in children and adolescents]. PMID- 11912775 TI - A pastor's perspective: getting through the grief. PMID- 11912776 TI - A social worker's assessment: tapping into resources. PMID- 11912777 TI - A nurse-ethicist's viewpoint: advocating for the family. PMID- 11912778 TI - A chaplain's reflection: bridging the gap. PMID- 11912779 TI - You gotta know there's gonna be a better tomorrow. Collaborating to comfort ventilator-dependent patients. PMID- 11912780 TI - The chaplain as spiritual facilitator. PMID- 11912781 TI - A clinic built on hope. PMID- 11912782 TI - Prescription for hope. How Esperanza began. PMID- 11912783 TI - Spiritual care. How the Christian community cared for mama. PMID- 11912784 TI - Bodywork. PMID- 11912785 TI - All the days of their lives. An eldercare alternative. PMID- 11912786 TI - Family ties. How to stay connected with the elderly. PMID- 11912787 TI - Facing reality: a team approach. PMID- 11912789 TI - Pyoderma gangrenosum: a case study for pain management in dermatology nursing. AB - The Joint Commission on Accreditation of Healthcare Organizations recently approved new standards for pain assessment and management. These standards will influence the practice of dermatology nursing in every setting. Nurses play a critical role in the care of patients with pain. Pyoderma gangrenosum is used as the basis for applying the new pain standards. PMID- 11912790 TI - A plan to promote the prevention and early detection of melanoma. AB - Malignant melanoma will affect millions of Americans in the year 2000 and its incidence is rising 7% per year (Skin Cancer Foundation, 1992). Numerous programs have been developed in various parts of the country to control the rate of increase, with mixed results. Public education programs can potentially prevent skin cancers and promote early detection of melanoma. An innovative prevention and detection program involving schools, pediatricians, nurses, and state agencies that will target those who are at risk for developing melanoma and skin cancer was developed by the author as a new mechanism by which the public can be educated regarding this disease. PMID- 11912791 TI - Evaluation and treatment of thermal injuries. AB - An estimated 1 to 2 million Americans suffer burn injuries each year. The majority of these are minor; however, approximately 70,000 individuals sustain injuries severe enough to require admission to a hospital or burn center for treatment. PMID- 11912792 TI - What's your assessment? AB - The "What's Your Assessment?" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11912793 TI - Eyelid dermatitis. AB - Eyelid dermatitis is a common problem, probably owing to the thinness of eyelid skin and to myriad chemical preparations that touch or are placed near the eyelids. These include eye makeups, ophthalmic preparations, airborne allergens, anti-aging medications, sun screening agents, and chemicals that touch the face from hand usage. Finding the cause starts with an excellent history followed by an exacting examination of the skin. PMID- 11912794 TI - Diagnosis and treatment of atopic dermatitis. PMID- 11912795 TI - Opportunities for learning about evidence-based practice. PMID- 11912796 TI - WAVE: a pocket guide for a brief nutrition dialogue in primary care. PMID- 11912797 TI - Nutritional modulation of blood parameters in type 2 diabetes. PMID- 11912798 TI - The role of the certified diabetes educator in telephone counseling. AB - PURPOSE: This paper describes a telephone-based, outcomes-focused approach to diabetes education provided by certified diabetes educators (CDEs). METHODS: Random chart audits were conducted to evaluate the scope of practice and effectiveness of telephone-based interventions provided by CDEs to people with diabetes. Four case studies and a sample prevention case are used to illustrate the role of telephone-based CDEs in providing diabetes education. RESULTS: Counseling provided by CDEs helped to identify potential barriers and strategies for making lifestyle behavioral changes. CONCLUSIONS: Telephone-based counseling is a brief, effective, ongoing intervention that gives patients with diabetes immediate access to CDEs who provide education to support lifestyle behavioral changes. PMID- 11912799 TI - Adherence to 1997 diabetes screening guidelines in a large ambulatory clinic. AB - PURPOSE: The purpose of this study was to evaluate whether the 1997 American Diabetes Association screening guidelines were being implemented by primary care providers. METHODS: A retrospective health record review was undertaken in a large midwestern ambulatory care clinic. A master list was developed of clients aged 45 to 54 years who had been given a physical examination between January 1, 1998, and June 30, 1998. A total of 310 records were systematically selected and abstracted. Data were analyzed using descriptive statistics. RESULTS: Although all subjects met the age criterion by design, and a high percent were high risk, only 57.7% were screened for diabetes. CONCLUSIONS: Adherence to the 1997 screening guidelines was inadequate in this practice setting; therefore the authors suggest replicating this study in other practice settings. Diabetes educators should continue to promote the practice of screening high-risk individuals of all ages. PMID- 11912800 TI - The impact of barriers and self-efficacy on self-care behaviors in type 2 diabetes. AB - PURPOSE: This cross-sectional, correlational study examined the relationships of diabetes-specific treatment barriers and self-efficacy with self-care behaviors. METHODS: A total of 309 people with type 2 diabetes participated in this study. All of the factors were assessed by self-report questionnaires. Self-care behaviors included exercise, diet, skipping medication, testing blood for glucose, adjusting insulin to avoid or correct hyperglycemia, and adjusting diet to avoid or correct hypoglycemia. RESULTS: Perceived barriers to carrying out self-care behaviors were associated with worse diet and exercise behavior. Greater self-efficacy predicted more frequent blood glucose testing, less frequent skipping of medication and binge eating, and closer adherence to an ideal diet. Nontraditional dimensions of self-efficacy were associated with worse self-care. Self-efficacy explained 4% to 10% of the variance in diabetes self care behaviors beyond that accounted for by patient characteristics and health beliefs about barriers. CONCLUSIONS: The findings of this study provided support for Rosenstock's proposal that a person's self-perceived capability to carry out a behavior should be incorporated into an expanded health belief model. PMID- 11912801 TI - Patient priorities and needs for diabetes care among urban African American adults. AB - PURPOSE: This study was conducted to determine diabetes care priorities and needs in a group of urban African American adults with type 2 diabetes mellitus. METHODS: One hundred nineteen African American adults with type 2 diabetes, aged 35 to 75, received behavioral/educational interventions from a nurse case manager, a community health worker, or both. Priorities and needs were assessed during 3 intervention visits. RESULTS: The most frequently reported priorities for diabetes care were glucose self-monitoring (61%), medication adherence (47%), and healthy eating (36%). The most frequently addressed diabetes needs were glucose self-monitoring and medication adherence. Most of the intervention visits (77%) addressed non-diabetes-related health issues such as cardiovascular disease (36%) and social issues such as family responsibilities (30%). CONCLUSIONS: Participants' self-reported priorities for diabetes care directly reflected the diabetes needs addressed. Needs beyond the focus of traditional diabetes care (social issues and insurance) are important to address in urban African Americans with type 2 diabetes. Interventions designed to address comprehensive health and social needs should be included in treatment and educational plans for this population. PMID- 11912802 TI - A summary of the certification examinations from 1999. PMID- 11912803 TI - Gestational diabetes mellitus: diagnosis, treatment, and beyond. PMID- 11912804 TI - Satisfaction with automated telephone disease management calls and its relationship to their use. AB - PURPOSE: This paper reports the results of an assessment of automated telephone disease management (ATDM) among patients in 2 randomized trials evaluating ATDM as an adjunct to usual care. METHODS: During the 12-month follow-up interview, 256 low-income adults with diabetes from the intervention groups of 2 randomized trials were asked to respond to questions about their satisfaction with the ATDM process. Variation in satisfaction was examined across groups, and satisfaction reports were correlated with the extent to which patients completed ATDM assessments and used them to report glycemic levels or access educational messages. RESULTS: Overall, 85% of patients reported that they were satisfied with the ATDM calls, 82% reported that they would be more satisfied with their health care if such calls were available to patients, and 76% reported that they personally would choose to receive such calls in the future. Most patients reported few difficulties completing ATDM assessments and found the calls to be a useful adjunct to their care. Some, however, found the calls intrusive; 16% reported that the calls were a bother. CONCLUSIONS: Patients were satisfied with ATDM calls as part of their diabetes care. Satisfaction played some part in determining patients' use of ATDM assessment and health education, although other barriers also contributed to less than optimal usage rates. PMID- 11912805 TI - A Web-accessible core weight management program. PMID- 11912806 TI - A survey of physician awareness of West Virginia's coverage for treatment of diabetes. PMID- 11912807 TI - Diabetes education outcomes: what educators are doing. PMID- 11912808 TI - National Diabetes Education Outcomes System: application to practice. PMID- 11912809 TI - Issues for the coming age of continuous glucose monitoring. AB - PURPOSE: This article reports the results of a symposium in which diabetes educators considered and discussed issues that are likely to arise when continuous glucose monitoring (CGM) becomes available and readily accessible. METHODS: Fifteen certified diabetes educators and 5 others with complementary expertise participated in a discussion based on their responses to 11 questions designed to elicit perspectives on issues related to CGM. Issues for discussion and debate include those related to patient acceptance and lifestyle, implications for professional practice and reimbursement, concerns about professional liability, use of CGM data by insurers and payers, and CGM data transfer. RESULTS: Educators offered varied and sometimes conflicting responses to CGM-related issues. CONCLUSIONS: Awareness of CGM-related issues will likely become an important part of diabetes professional development and perspectives in practice. Identifying and framing the issues before the new technologies become available allow diabetes educators to participate proactively in structuring the emerging policies, procedures, and standards of care. PMID- 11912810 TI - Identifying variables associated with inaccurate self-monitoring of blood glucose: proposed guidelines to improve accuracy. AB - PURPOSE: This study was conducted to evaluate patients' proficiency in self monitoring of blood glucose (SMBG). METHODS: Diabetes nurse educators in 4 suburban Minneapolis clinic sites surveyed the SMBG training/cure practices of 280 patients with type 1 and type 2 diabetes. Participant SMBG technique was measured by direct observation. Participants performed a finger puncture and used their own meters to measure the first blood sample. A second sample was measured on the HemoCue B Glucose analyzer, and a third sample was used to measure hemoglobin. The series of tests were then repeated. If either of the 2 glucose tests was more than 15% from the HemoCue value, participants were reeducated about the manufacturer's suggested procedure. RESULTS: Of the 280 participants, 19% had blood glucose test results greater than the 15% limit for meter accuracy. After reeducation, 69% of those who had initially failed achieved acceptable results. The most significant problems were lack of periodic meter technique evaluation, difficulty using wipe meters, incorrect use of control solutions, lack of hand washing even when observed, and unclean meters. CONCLUSIONS: As a result of the study, guidelines were subsequently developed to evaluate meter accuracy in an outpatient setting. Further effort is needed to establish standards for evaluating SMBG. PMID- 11912811 TI - Using a primary nurse manager to implement DCCT recommendations in a large pediatric program. AB - PURPOSE: This study was conducted to determine whether recommendations from the Diabetes Control and Complications Trial (DCCT) could be implemented in a large pediatric population using a diabetes clinical nurse specialist program coordinator dedicated to intensive management. METHODS: Patients' charts were reviewed to examine HbA1c levels from before the results of the DCCT were published and again 1 year after the recommendations were implemented. Patients who met the following criteria (N = 124) were enrolled: type 1 diabetes, less than 18 years old, followed at Yale for 1 year prior to the results of the DCCT and 1 year after, and HbA1c level recorded in the medical chart. RESULTS: HbA1c levels were significantly lower 1 year after implementing the DCCT protocol; 3 years later these same patients improved even further as evidenced by another decrease in HbA1c levels. The patients were taking more insulin (more Ultralente and regular insulin and less NPH) and had an increased number of injections at both the 1-year and 3-year follow-up points. CONCLUSIONS: The DCCT protocol can be implemented in a large population of pediatric patients with the help of a nurse who is dedicated and available to the patients for ongoing adjustments and provides creative ways to overcome the barriers to achieving normoglycemia. PMID- 11912812 TI - The effect of diabetes self-management education with frequent follow-up on the health outcomes of African American men. AB - PURPOSE: This study was conducted to determine whether objective clinical, patient performance, quality-of-life, and subjective outcomes are significantly different among African American men with type 2 diabetes who received follow-up at either monthly or 3-month intervals after participating in a structured diabetes self-management education program. METHODS: Prior to the diabetes self management education program, 30 African American men with type 2 diabetes were randomly assigned to 2 groups to receive telephone follow-up at either monthly or 3-month intervals over a 6-month period. Information obtained at follow-up contact included HbA1c level, perception of general health, and present diabetes knowledge. In addition, daily foot care, dietary, exercise, and medication compliance measures were assessed postprogram. RESULTS: There were no significant differences between the participants who received follow-up at monthly and 3 month intervals on any measures of the selected clinical, patient performance, quality-of-life, and subjective outcomes. CONCLUSIONS: This cross-sectional study showed that telephone follow-up at 3-month intervals following a structured program of diabetes self-management education may be just as effective in contributing to favorable diabetes health outcomes as monthly follow-up. PMID- 11912813 TI - Elevated plasma glucose levels increase risk for complications. AB - Diabetes is a chronic health condition that requires tremendous energy and care throughout the patient's lifespan. Clinically, it is easier to prevent glucose excursions than to treat them once they occur! There are data to suggest that control of blood glucose levels will prevent damaging glucotoxicity, preventing or delaying diabetes-related complications. New treatment goals of the American Diabetes Association are the following: Fasting plasma glucose levels of 80 to 120 mg/dL; Initial treatment of diet, self-care management, education, and exercise; Additional treatment recommended at FPG > 140 mg/dL; Goal HbA1c levels < 7%, with treatment changes recommended at > 8%; Therapy with either insulin secretagogues or insulin sensitizers if poor glycemic control persists after 2-3 months. The clear message to both patients and diabetes care providers is that aggressive and early therapeutic intervention in type 2 diabetes is the key to improving and maintaining continued glycemic control and preventing micro- and macrovascular complications. PMID- 11912814 TI - Treatment strategies and new therapeutic advances for type 2 diabetes. AB - Because type 2 diabetes is caused by two defects, impaired insulin secretion and insulin resistance, logical management of diabetes will include combination therapies to treat this dual condition. Initial combination therapy should include an insulin secretagogue and an insulin sensitizer, with the addition of insulin in the evening if the HbA1c remains greater than 8%. Treatment to target should be clearly defined to achieve HbA1c < 7% unless there are specific individual considerations that make higher HbA1c levels acceptable or desirable. Patients are now treated earlier, when fasting blood glucose levels are in the 126 to 140 mg/dL range; and drugs with less chances of hypoglycemia are preferred at this stage. However, low-dose combination therapy as an early initial treatment, if HbA1c remains > 7%, is an emerging aggressive strategy that requires further consideration and further studies to prove its long-term efficacy and safety. PMID- 11912815 TI - Pathophysiology of type 2 diabetes and its relationship to new therapeutic approaches. PMID- 11912816 TI - Implications of the United Kingdom Prospective Diabetes Study (UKPDS) results on patient management. AB - The goal of treatment of type 2 diabetes is to treat early and balance the risk/benefit ratio of aggressive therapeutic intervention for the patient. The results of the UKPDS trial have shown that treatment to improve overall glucose control significantly reduces the risk of microvascular complications and thereby improves the quality of life for persons with diabetes. Optimal medical management includes a team approach and uses continuous communication and education, medical treatment, lifestyle modifications, and behavior changes to improve glycemic control. PMID- 11912817 TI - A patient-centered approach to nurse orientation. AB - An orientation pathway was developed using a patient-centered approach. The pathway provides a guide or "road map" for the preceptor and new nurse to care for high-volume patient populations in medical and surgical units. Benefits include application of the nursing process, promotion of critical thinking skills, reduction of reality shock, and improvement in job satisfaction and retention. This article describes the rationale behind the approach and its application to nurse orientation. Steps in the process are described. Staff development educators can use the steps to develop orientation pathways for other practice settings. PMID- 11912818 TI - Problem-based learning in clinical practice facilitating critical thinking. AB - Problem-based learning has established a strong reputation within education programs delivered to a wide variety of healthcare professionals. Nevertheless, most accounts of problem-based learning relate to classroom settings, where issues of patient safety and competing demands upon time are not a primary concern. The authors explore ways in which problem-based learning may be utilized within clinical practice to enhance the professional development of nurses. A framework for thinking about problems and professional responses is outlined, and illustrations are drawn from the maternal-child health practice setting. PMID- 11912819 TI - Physical restraint management of hospitalized adults and follow-up study. AB - During the winter of 1998 the management of 118 (N = 118) physically restrained adult patients in a 238-bed urban acute care hospital were assessed by 26 registered nurse (RN) data collectors. In the spring of 1999, following a comprehensive hospital-wide staff development program and revised physical restraint protocol, 10 RN data collectors conducted a follow-up study of 53 (N = 53) restrained adults in the same institution. In both studies, data regarding restraints management were gathered using a Restraint Management Improvement Indicator. Following a program of restraint management education, substantial improvements were found for virtually all of the physical restraint indices studied. The findings suggest that future educational efforts should be undertaken to further improve the documentation in hospital medical records regarding medical orders and ongoing observation, assessment, and interventions for physically restrained patients. Future research should further document and study interventions to reduce or eliminate the use of physical restraints. PMID- 11912821 TI - Educating to manage the accelerated change environment effectively: Part 1. AB - Without appropriate educational preparation, nurse managers may not have the competencies to effectively manage accelerated change that is so pervasive on the front lines of healthcare organizations in transformation. Part 1 of this article describes the findings from an extensive literature review of nursing, business, and higher education literature on the subject of change management. A comprehensive reference list is provided. Part 2 reports the results of a Delphi study from which baccalaureate-prepared nurse manager experts and nurse educator experts in baccalaureate nursing programs validated what linear and nonlinear change management concepts they believed were relevant in managing change in today's dynamic environment. With key roles as organizational change agents, staff development educators and administrators can use the validated concepts to develop educational offerings to promote effective change management. PMID- 11912820 TI - Comparing nutrition knowledge exam scores with reported nutrition topics of interest among nursing home nurses. AB - Nursing home nurses (licensed practice nurses and registered nurses) were assessed to determine nutrition training issues that have a potential impact or influence on protein-calorie malnutrition of residents. The results of a 50-item nutrition knowledge exam are reported and compared with what nurses report they are interested in knowing about nutrition. Nurses in nursing homes were found to lack sufficient nutrition knowledge to meet dietary needs of elderly residents. PMID- 11912822 TI - Measurement of return on investment of workplace education: an annotated list of references. AB - In the context of downsizing and with the decline of revenue in healthcare organizations, educators recognize the need to develop strategies to measure the subsidy of staff development programs to the organization's welfare. An annotated reference list is provided to assist educators who have little time for an extensive literature review with a place to begin development of a plan to measure return on investment. PMID- 11912823 TI - Using games to determine learning styles. PMID- 11912824 TI - Reducing wrinkles and other signs of aging. AB - As the aging population increases so does the demand for more effective modalities to combat the ravages of skin aging. It is important for dermatology nurses to understand and be able to explain to patients the preventive measures for skin aging and what can be realistically expected from currently available treatments. PMID- 11912825 TI - Early detection and treatment of melanoma: update 2000. AB - Risk factors for the development of melanoma, early detection, biopsy technique, and treatment recommendations are discussed. Survival depends upon the stage at the time of diagnosis; thus, early detection of thin melanoma limited to the skin's surface enhances survival. PMID- 11912826 TI - What's your assessment? PMID- 11912827 TI - Narrow-band UVB: a practical approach. AB - Narrow-band UVB therapy offers a new and exciting treatment option and area of study in phototherapy. The narrow-band skin type protocol offers a safe and effective treatment plan for those who do not routinely use, or wish to use, minimal erythema dosing to establish starting doses for narrow-band UVB therapy. PMID- 11912828 TI - Emulsifying oil in the bath helps children with Eczema. PMID- 11912829 TI - Wound assessment and evaluation. AB - The "Wound Assessment and Evaluation" series includes a short case presentation and differential diagnosis. It is followed by a discussion of the disease or condition and the rationale used in each step of the assessment. PMID- 11912830 TI - Camouflage therapy. AB - Camouflage cream was introduced over 50 years ago to assist in the rehabilitation of severely burned pilots injured during World War II. Today, camouflage therapy can be used to conceal discoloration such as postoperative bruising and erythema. Pre and postoperative teaching regarding camouflage therapies are very important parts of the healing process, and dermatology nurses can play a vital role in the teaching and healing process. PMID- 11912831 TI - [Meibomian adenocarcinoma of the free palpebral edge. Report of a case]. AB - A case of meibomian carcinoma of the left eyelid is reported in a 72-year-old female patient. The tumor had been present on the left eyelid for months. Clinically, the tumor appeared as a reddish mass implanted on the external part of the free margin of the left superior eyelid. An excisional biopsy disclosed meibomian carcinoma. A total resection of the left superior eyelid was followed by plastic surgery. Results after a one-month follow-up were very satisfactory. This case is emphasizes the importance of an early diagnosis which enabled us to perform a rather conservative treatment limited to the removal of the affected eyelid. The diagnosis of meibomian carcinoma is infrequent but it must be kept in mind in cases of tumor without the typical clinical characteristics of a basal cell or squamous cell carcinoma. Complete removal surgery may bring a curative effect and histopathology has a key role in the diagnosis of meibomian carcinoma. PMID- 11912832 TI - [Botulinum toxin in ophthalmology]. PMID- 11912833 TI - [Precision and reliability of Orbscan and ultrasonic pachymetry]. AB - PURPOSE: To compare the accuracy and reproducibility of the orbscan pachymetry and ultrasonic pachymetry in the normal eye and in the penetrating keratoplasty eye. METHODS: Pachymetric measurements were assessed in 50 eyes of 25 normal patients and 50 eyes of 48 patients who had undergone penetrating keratoplasty using both Orbscan II and ultrasonic pachymetry (Tomey SP-2000). For each eye, 2 successive measures were recorded with both instruments. For both devices, the default setting was used. Orbscan pachymetry maps were divided into 5 groups using a modification of Liu's classification. RESULTS: Orbscan pachymetry strongly correlated with ultrasonic pachymetry (rs = 0.91; p < 0.001). Ultrasonic pachymetry values and Orbscan pachymetry values showed no significant differences in the normal group (respectively, 557 microns +/- 36 and 555 microns +/- 34; p > 0.05). Ultrasonic pachymetry values and orbscan pachymetry values were significantly different in the penetrating keratoplasty group (respectively, 571 microns +/- 52 and 550 microns +/- 54; p < 0.001). The best value for the acoustic factor for Orbscan pachymetry in the penetrating keratoplasty group was 0.89. Ultrasonic pachymetry reproducibility and Orbscan pachymetry reproducibility were not significantly different (0.86% +/- 0.61 v. 0.67% +/- 0.63; p = 0.13 in the normal group; 1.22% +/- 0.81 v. 1.23% +/- 1.13; p = 0.92, in the penetrating keratoplasty group). Both pachymetry methods showed less reproducibility in the penetrating keratoplasty group than in the normal group (p < 0.02). Thinnest point localization was significantly different in both groups (p < 0.001). In 66% of the normal group, the thinnest point of the cornea was located in inferotemporal quadrant. This point was located at an average of 0.63 +/- 0.25 mm from the visual axis in the normal group and 1.60 +/- 0.81 mm in the penetrating keratoplasty group (p < 0.001). Whereas "Centered round" (40%) and "centered oval" (34%) were the most common patterns in the normal group, "decentered oval" (40%) and "irregular" (30%) were more frequent in the penetrating keratoplasty group (p < 0.001). CONCLUSION: Orbscan pachymetry strongly correlated with ultrasonic pachymetry. Reproducibility of both methods is excellent and not significantly different. Thinnest point localization and pachymetric map classification are significantly different in normal eyes and in penetrating keratoplasty eyes. PMID- 11912834 TI - [Optic biometry in intraocular lense calculation for cataract surgery. Comparison with usual methods]. AB - PURPOSE: To compare axial length and intraocular lens power calculated from three biometry methods, then to study refractive postoperative results to assess the predictive value of each method. MATERIAL AND METHODS: This prospective study included 40 eyes planned for cataract surgery. Two skilled operators participated in this study: One for the surgery and the other for the biometry and measurement of intraocular lens power. For intraocular lens power, we used the optic biometer from Zeiss and the echograph B Ultrascan from Alcon. IOL power calculation was performed using the usual mathematical formulas based on 3 biometry methods. 1- keratometry measurement, anterior chamber depth (ACD), and axial length using optical biometry; 2--keratometry measurement using the Javal keratometer and biometry using the B mode ultrasonography; 3--keratometry measurement using the Javal keratometer and biometry using A mode ultrasonography. RESULTS: The average age of our patients was 69.5 years old, ranging from 52 to 81 years old. The average axial length was 23.46 mm with, ranging from 20 to 32.73 mm. The average keratometry with optic biometry was 43.97 diopters +/- 1.44 versus 43.84 diopters +/- 1.45 with the Javal keratometer. 40 eyes were examined and there were 4 failures (10%) for axial length measurement by optic biometry because the cataract was very dense. Biometric preoperative results with the 3 methods show that there was a statistically significant difference between the A mode and the B mode optic biometry (P < 0.006). On the other and, there was no statistical difference between optic biometry and the B mode. CONCLUSION: Optic biometry has a number of advantages. This is new method, is non invasive, easy to use, with no contact, and it is reliable. Results with this method are more precise than with ultrasonic biometry. For high myopia, optic biometry is a very valuable method. Its limits are total cataract and intraocular opacities; in these cases ultrasonic biometry is the best method. PMID- 11912835 TI - [Frequency of retinal changes in myopia in a black population]. AB - PURPOSE: Retinal detachment is the main complication of peripheral retinal lesions in myopia but seems rare in black people. We analysed peripheral retinal changes in myopia in a black population. MATERIAL AND METHODS: This study was conducted by a single ophthalmologist. The tests included a refraction, a retinal biomicroscopy (with a Volk lens and a 3-mirror Goldmann lens) and an indirect ophthalmoscopy. The results were compared by a chi square test. RESULTS: 50 people (100 eyes) were available for examination. The mean age was 28.21 years, the two extremes being at 11 and 54 years, with 68% females and 32% males. In 50 eyes the spheric equivalent was between -1 and -5.75 diopters (group I). In 50 eyes the spheric equivalent was greater than or equal to -6 diopters (group II). Snow was found in 16% of cases in group I and 34% of cases in group II (p = 0.0366). Tears were found in 2% of cases in group I and 12% of cases in group II (p = 0.116). Lattice degeneration was found only in group II. CONCLUSION: Retinal tear frequency was similar to the results of other authors. Comparative studies are necessary to evaluate the risk of retinal detachment in black populations. PMID- 11912836 TI - [Lamellar keratoplasty with air or viscoelastic substance injection]. AB - PURPOSE: To analyze the results of very deep lamellar keratoplasty using dissection with air and a viscoelastic substance. METHODS: This was a prospective monocentric noncomparative study. Candidates for lamellar keratoplasty were enrolled between November 1998 and July 2000. Deep lamellar dissection was performed following air injection into the cornea to create a white emphysema of the stroma. The dissection was performed to the Descemet membrane. Whenever a large bulla was present in the recipient bed, the dissection of the deepest stromal lamellae was performed by injecting a viscoelastic substance into the bulla. A full-thickness allogenic corneal button was sutured to the recipient bed. RESULTS: Fifteen eyes of 14 patients (mean age, 39.3 years) underwent deep lamellar keratoplasty: keratoconus (11 eyes), atopic keratoconjunctivitis (1 eye), herpes zoster keratitis (1 eye), corneal scar after pterygium surgery (1 eye), and rosacea keratitis (1 eye). Excluded from the analysis of the refractive outcome were patients who underwent intraoperative perforation (n = 3) and the patients with postoperative complications affecting the central visual axis: (n = 2 [hemorrhage in the interface and herpetic simplex keratitis]). The mean preoperative visual acuity was 0.10 (range, 0.05 to 0.3). After a 3.8-month follow-up, the mean best corrected visual acuity was 0.21 (range, 0.1 to 0.6). The visual results were better in patients with keratoconus (mean best corrected visual acuity: 0.22; range, 0.1 to 0.6). The mean postoperative astigmatism was 4.15 diopters (range, 0 to 8). CONCLUSION: Intrastromal air and viscoelastic substance injection appeared to be very useful for performing a very deep lamellar keratoplasty. The results of the refractive outcome were encouraging. Deep lamellar keratoplasty is an interesting alternative to penetrating keratoplasty, because it cannot induce progressive primary graft failure and allogenic endothelial graft rejection and it obviates the need to perform a lamellar dissection of the donor button. PMID- 11912837 TI - [Transpupillary thermotherapy in the treatment of choroid melanoma]. AB - PURPOSE: To report the results of primary transpupillary thermotherapy (TTT) for selected posterior pole choroidal melanomas. MATERIAL AND METHODS: Prospective non randomized study including 34 patients with choroidal melanoma treated with TTT using near-infrared radiation (810 nm) delivered from the diode laser. All treated tumors had either documented growth or clinical risk factors for future growth and/or metastasis. The treatment was delivered using a specially modified infrared diode laser through a slit lamp adaptor. A contact lens was placed on the cornea to view the fundus and focus the laser beam. Treatment was initiated using a 60-second exposure and a low energy level at 300 mW with a 3.0 mm beam width. The energy was raised stepwise by 50 to 100 mW until the surface of the tumor develop a light grayish discoloration. The TTT sessions were delivered at 3 month intervals. RESULTS: Among these 34 patients with choroidal melanoma, 29 patients presented with primary choroidal melanoma. Five tumors (15%) were late recurrences after conservative treatment. The mean initial tumor basal diameter was 7.2 mm and tumor thickness was 2.9 mm. Seventeen tumors (50%) touched the optic disc and 10 (29%) were under the fovea. After a mean of three treatment sessions and 20 months of follow-up, the mean tumor thickness gradually decreased to 2.3 mm at month 3 and 2.0 mm at month 6 after the initial TTT. The percent reduction of tumor thickness was 20% at month 3 and 29% at month 6. The mean final thickness was 1.7 mm. Treatment was successful in 33 patients (97%). Tumor regrowth was documented in one patient (3%) and required plaque radiotherapy. After treatment, visual acuity was the same or better than the pretreatment visual acuity in 21 eyes (62%) and worse in 13 eyes (38%). Intraocular complications included retinal traction in 12 eyes (34%) and vascular occlusion in 3 eyes (9%). CONCLUSION: This series confirms the efficacy of transpupillary thermotherapy in the management of selected posterior pole choroidal melanomas. Longer follow-up is still required to assess late local recurrence and the impact on metastatic disease. PMID- 11912838 TI - [Weill Marchesani syndrome. Report of a case]. AB - Weill Marchesani syndrome is a congenital disease that combines microspherophakia and skeletal abnormalities. The authors report a 19-year-old male, born of a consanguineous marriage, with a progressive decrease in visual acuity. The general examination showed a squat look, dwarfism, muscle hypertrophy, short hands and feet, and joint stiffness. The ophthalmological examination showed that visual acuity was limited to hand motion in the right eye despite correction and no perception of light in the left eye. Intraocular pressure was 36 mmHg in the right eye and 40 mmHg in the left eye. The anterior chamber was irregular with iridophakodonesis and microspherophakia of both lenses. The zonula was partially ruptured in the right eye. The iridocorneal angle was narrow. Fundus eye examination showed a pale optic disc with an excavation of 9/10 on the right. In the left eye, the optic disc was totally excavated. Cardiovascular check-up revealed rheumatic aortic valvular cardiopathy. The therapy consisted of combined surgery: phakophagia with anterior vitrectomy plus trabeculectomy operated on the right eye. Weill Marchesani syndrome is a rare congenital affection with a recessive autosomal transmission. The visual prognosis is dominated by secondary glaucoma due to pupillary blockage by the mobile eye lens. This observation illustrates the seriousness of spontaneous progression in Weill Marchesani syndrome, justifying the necessity of lens extraction before the onset of complications. PMID- 11912839 TI - [Proton therapy of occult neovessels in age-related macular degeneration]. AB - INTRODUCTION: The interest of radiation therapy in the management of age-related macular degeneration inaccessible to photocoagulation is still controversial. Our purpose was to demonstrate the feasibility and the possible efficacy of a single dose delivered to the macular region using a 65-MeV proton beam. MATERIAL AND METHODS: A phase II trial was set up using the cyclotron in Nice, France. Fifty eight patients were included after signing an informed consent. All patients presented with occult subfoveal choroidal neovascularization. A single dose of 9.1 Gy (i.e., 10 Gy cobalt equivalent) was delivered to the macular region. RESULTS: The results were analyzed 3, 6, 12 and, 18 months after proton therapy. At 3 months, the visual acuity was stable or enhanced for 86% of patients, at 6 months for 82.3%, at 12 months for 80%, and at 18 months for 61%. For 22 patients at follow-up at 18 months, the reasons for a decrease in visual acuity were a macular hemorrhage for 4 patients and a progression of the neovascular membrane for 3 patients. No secondary effects related to the treatment have been observed. Regarding the lesions visible on the angiographies (i.e., hemorrhage, exudates, subretinal detachment), we observed a stabilization or a decrease in two-thirds of the cases. CONCLUSION: Preliminary results of single-dose proton therapy are at least comparable to those obtained by other teams. A second study is in progress comparing 3 dose levels, looking for a dose-effect relationship. Furthermore, a randomized study comparing a single proton dose to a placebo will be necessary to assess the long-term value of proton treatment. PMID- 11912840 TI - [Retinal vascular occlusion and primary antiphospholipid syndrome. Report of 2 cases]. AB - INTRODUCTION: The antiphospholipid syndrome is characterized by the association of antiphospholipid antibodies and occlusive vascular events. The ocular vascular damage is described in the literature and may reveal the syndrome. It's diagnosis is important because this disease, which generally affects young people, may endanger ocular and vital prognosis. Two cases are reported. PATIENTS: Two young people aged 22 and 32 presented with a sudden unilateral loss of vision. In both cases, fundus examination and fluorescein angiography revealed a severe retinal vascular occlusion. For one patient, an antiphospholipid syndrome was discovered during etiological check-up. for the second one, it was associated with a heterozygote state mutation of Leiden V factor. In both cases the visual function of the affected eye was lost. DISCUSSION: In most cases central artery or vein occlusion means atherosclerosis. However, other etiologies must be studied in young patients. According to several studies: antiphospholipid syndrome has been detected in 5% to 33% of the patients showing a major vascular retinal obstruction. The association of thrombophilia must be considered because it increases the risk of thrombotic recurrence. CONCLUSION: The antiphospholipid syndrome must be studied in cases of numerous and severe retinal vascular obstruction occurring in young patients, even if vascular risks exist. This diagnosis is important because it may imply a long-lasting anticoagulative or an antiaggregative treatment to significantly reduce the risk of recurrent thrombotic events. PMID- 11912841 TI - [A familial case of chronic progressive external ophthalmoplegia associated with mitochondrial disease]. AB - Mitochondrial myopathies are rare hereditary diseases that affect the energy functions of the mitochondria. Clinical manifestations are variable and sometimes multisystemic. Progressive external ophthalmoplegia constitutes the most frequent clinical form. Unfortunately, the diagnosis and the treatment of these mitochondrial abnormalities stay, today, even difficult. We report ophthalmic findings and the course of the disease in members of a family with chronic progressive external ophthalmoplegia presenting with severe acquired blepharoptosis. From study at the family background, the inheritance seemed autosomal dominant. In one case, a comprehensive workup, including muscular biopsy and molecular genetics disclosed a mitochondrial myopathy. During the 30 year follow-up, the patients were operated on for their ptosis several times, because of recurrences and uneven results. PMID- 11912842 TI - [Lisch nodules: description of 2 clinical cases and their significance]. AB - We report two cases of Lisch nodules in both eyes in two women. It was the only ocular manifestation of their Von Recklinghausen disease. The other general signs of their disease were cutaneous neurofibromas. Lisch nodules are melanocytic hamartomas consisting of melanocytic cells containing various quantities of pigment. They can be found in 90 to 100% of neurofibromatosis patients over 6 years of age. They are practically pathognomonic of type 1 neurofibromatosis and their finding is an important diagnostic argument. They must be distinguished from other iris nodules: nevus, melanoma, inflammatory nodules, and development anomalies. PMID- 11912843 TI - [Cytomegalovirus retinitis despite normal CD4 levels in an HIV patient. Report of a case]. AB - We report the case of a 54-year-old HIV-seropositive man who was referred to us with unilateral uveitis. The patient had been taking triple antiretroviral therapy (three reverse transcriptase inhibitors) for one week when he presented with blurring of vision in the left eye. Two weeks later, active cytomegalovirus retinitis was suspected after fundus examination and ganciclovir was immediately administered. Even if the CD4 cell count was normal (423/microliter), the diagnosis was confirmed by the presence of CMV in the anterior chamber. This case shows that an elevated CD4 count is not incompatible with CMV retinitis, especially at the beginning of the antiretroviral therapy. PMID- 11912844 TI - [Coats disease]. AB - In spite of an unclear definition of Coats disease, this angiomatosis can be defined by the presence of retinal telangiectasis of nondetectable etiology, complicated by severe exudative phenomena. The diagnosis of this serious affection remains difficult because of its numerous clinical forms, which vary according to the age at which the disease appears and the progressive nature of the exudative phenomena. The consequences on the young child are all the more dangerous because it is too often discovered late. It seems necessary to attempt to stop the progression of Coats disease by destroying the telangiectasis responsible for intra- and subretinal exudation. No coagulation technique has been shown to be more effective than others and the predominance of heterogeneous series in the literature makes it difficult to evaluate the therapeutic results. In cases of severe retinal detachment, these coagulations can only be done after the often difficult drainage of the subretinal fluids. PMID- 11912845 TI - [Eales]. AB - Eales' disease, first described by Eales Henry in 1880, is an idiopathic obliterative vasculopathy that primarily affects the peripheral retina of adults. From nosologic point of view, we could distinguish the "true Eales disease" affecting males in India and portions of the Middle East and the "Eales' like disease" with no geographical or sex predisposition. Vascular sheathing, peripheral retinal nonperfusion and neovascularization are the principal clinical and angiographic findings. It remains a diagnostic by exclusion and retinal diseases with other causes of inflammation or neovascularization must be excluded. In the absence of a specific treatment, retinal photocoagulation to the nonperfused retina and vitrectomy for recurrent vitreal hemorrhage could be proposed. PMID- 11912846 TI - [Retinal involvement in hemoglobinopathy]. AB - Sickle cell disease is the most common and severe hemoglobinopathy. Ocular complications are related to ischemic retinopathy. Retinal examination has to be done in homozygous or double heterozygous patients and when the sickle trait is present with additional systemic vascular conditions. Fluorescein angiography is the main investigation. Photocoagulation to ischemic areas is effective in achieving regression of neovascularization. Various measures to reduce ischemia are recommended for surgical procedure. Multidisciplinary teams should take on severe cases. PMID- 11912847 TI - [Radiation-induced retinopathy]. AB - Radiation retinopathy is observed after intraocular tumor irradiation (mainly uveal melanomas, retinoblastoma being more seldom) or intracranial tumours radiotherapy. Today radiation retinopathy is also seen after irradiation for choroidal neovascularization observed in age-related macular degeneration. Radiation retinopathy after posterior uveal tumor irradiation will be described. Besides biomicroscopy, fluoresceinic angiography is useful for diagnosis more than treatment which is without efficacy in most cases. Major radiosensitivity of vascular endothelium due to a rapid cell turn over could explain not only radiation maculopathy but also optic disc neuropathy. Radiation doses and tumour location are mainly responsible for macular and/or optic nerve head vascular complications. Moreover, after vascular and tissular retinal complications, choriocapillaris and main choroidal vessels can also be affected after radiotherapy. PMID- 11912848 TI - The CEFP as a model for integrating evaluation within organizations. AB - This article identifies and discusses the innovative contributions of the Collaborative Evaluation Fellows Project (CEFP) as a model for evaluation training and organizational development. The importance of the CEFP model to the effectiveness and accountability of nonprofit organizations is best understood within the larger context of the historical emergence of evaluation as a specialized field of professional practice and the documented challenges of integrating evaluation within organizations. Key elements of the model discussed include a strong commitment to evaluation use (utilization-focused evaluation); careful attention to evaluation standards; developing core evaluation competencies among fellows; supporting collaboration among evaluation fellows, facilitators, and faculty; working to infuse evaluative logic, values, and thinking into the planning and management processes of the American Cancer Society; modeling effective evaluation practice by intensively evaluating its own effectiveness, both internally and with external reviewers; and using the findings to incorporate lessons learned, overcome barriers, and adjust to emergent challenges. PMID- 11912849 TI - Integrating theory and practice. Conceptual frameworks of the CEFP. AB - In this article, we describe how different conceptual frameworks contributed to the design, development, and implementation of the Collaborative Evaluation Fellows Project (CEFP). These frameworks are the following: utilization-focused evaluation (Patton, 1997); evaluative inquiry for learning in organizations (Preskill & Torres, 1999); and collaboration for a change (Himmelman, 1994). In addition, we explain how The Program Evaluation Standards (Joint Committee on Standards for Educational Evaluation, 1994) provided criteria for implementing and assessing the evaluation studies conducted through the CEFP. We conclude the article with examples of how the CEFP has changed the way American Cancer Society staff members and volunteers think about and conduct evaluations. PMID- 11912850 TI - Evaluation of triple touch. An assessment of program delivery. AB - Triple Touch is a breast health education program developed by the Florida Division of the American Cancer Society in 1994. The program reaches more Florida women each year, and has a larger group of volunteers, than any other breast cancer program sponsored by this American Cancer Society division. The purpose of the evaluation of Triple Touch was to assess and evaluate the program delivery methods of the program. The specific objectives were to identify and describe instructor characteristics, program delivery patterns, and satisfaction with the program. Results will be used for the following purposes: 1) for the future development and implementation of the Triple Touch program; and 2) for building greatly needed research-based knowledge on program implementation and delivery methods. Last, important lessons were learned from using the Collaborative Evaluation Fellows Project model to conduct evaluation research from all perspectives, including those of the evaluation fellow, the evaluation faculty, and the evaluation facilitator. PMID- 11912851 TI - An evaluation of I Can Cope. Insights into program design and implementation. AB - The I Can Cope program is an educational program developed to provide information on how to promote physical and emotional well-being to individuals with cancer, their families, and friends. Through focus group and individual interviews and a mailed survey, the effectiveness of the selection, marketing, implementation, and support processes associated with implementing the program in the central Arizona and greater Las Vegas, Nevada, regions was evaluated. Findings, recommendations, lessons learned, and use of findings are discussed. PMID- 11912852 TI - Doing Our Part So Kids Don't Start. An evaluation of a statewide public awareness campaign. AB - This article describes outcomes related to the product and process involved in an evaluation of one component of a statewide public awareness campaign, the Doing Our Part So Kids Don't Start toolkit. The campaign was designed to garner support for and to increase compliance with a recent Minnesota law intended to reduce youth access to tobacco. More than 35,000 toolkits were distributed statewide. The results of the evaluation suggest that the toolkit was successful in reaching and engaging its target audience and should be considered a good strategy for disseminating information to a wide variety of individuals, particularly those new to tobacco control, and for encouraging them to participate in preventing tobacco use among youth. Future American Cancer Society endeavors would benefit from considering the needs of both the program planner and program evaluator before implementing new projects or programs. PMID- 11912853 TI - The Collaborative Evaluation Fellows Project. Background and overview of the model. AB - The Collaborative Evaluation Fellows Project (CEFP) of the American Cancer Society builds organizational capacity for evaluation by partnering with schools of public health to conduct local evaluation studies. This model was envisioned in response to the American Cancer Society's growing demand for program evaluation and the challenges of institutionalizing evaluation nationwide and at all levels of the organization. The CEFP also formalizes partnerships between the American Cancer Society and universities, and provides practical opportunities for public health graduate students and faculty. Working with an American Cancer Society evaluation facilitator and an advisory group, faculty and graduate students use a 15-step process to conduct evaluation studies at the local level. Students also can use these data for their master's these. The 15-step process is based on the utilization-focused evaluation approach and is designed to foster the use of evaluation findings by program stakeholders. The CEFP model has been implemented in American Cancer Society offices nationwide and has been replicated with another nonprofit health organization. PMID- 11912854 TI - Evaluation of the American Cancer Society's Research Promotion Guide. AB - The American Cancer Society has supported cancer research for more than 50 years and has devoted more than $2.2 billion to cancer research. This article describes an evaluation of the Research Promotion Guide of the American Cancer Society, a reference tool developed by the National Home Office for regional division staff and volunteers whose work involved public relations and fundraising. The purposes of this study were the following: 1) to determine the level and type of use of the guide; and 2) to assess factors that may influence the level and type of use. Fifty-five participants were interviewed by telephone. Overall, 78% of them were aware of the guide. Three factors significantly and positively associated with level of use were: compatibility (the perception that the guide is relevant to one's job); attending a training session; and years employed at the American Cancer Society. This study, a pilot project for the Collaborative Evaluation Fellows Project (CEFP), demonstrated that collaboration between nonprofit and academic institutions is feasible and can serve the needs of the students and organizations. The lessons learned can be applied to evaluations in general and to future CEFP projects. PMID- 11912855 TI - Investigation of healthcare providers' patterns of referring breast cancer patients to Reach to Recovery. AB - Providers who referred patients to Reach to Recovery (Reach), an American Cancer Society breast cancer support group, were compared with those who did not to evaluate whether providers who identified problems with the program were less likely to make referrals. Also considered were contact with a Reach volunteer, having a Reach program in the area, years since residency or medical training, perceived value of the Reach program, belief that one's peers refer patients to Reach, urban or rural location of practice, and size of practice. When other variables were considered, physicians who identified problems with Reach were no less likely to refer patients to the program than those who did not. The factors most associated with referral (P < .001) were "having a Reach program in the area" and "having had contact with a Reach volunteer." Implications of these findings and the experience of conducting the evaluation through the Collaborative Evaluations Fellows Project are discussed. PMID- 11912856 TI - Local and national uses of a Road to Recovery evaluation. AB - Evaluation fellows from the George Washington University School of Public Health and Health Services conducted an evaluation of the Road to Recovery program of the Mid-Atlantic division of the American Cancer Society. The evaluation included qualitative analysis of program operation, mailed surveys, in-depth interviews with patients and drivers, and interviews with social workers from treatment centers. Results indicated that patients and drivers were satisfied with the program. Patients appreciated the ability of drivers to provide personalized, reliable service. The recruitment of sufficient drivers to meet transportation demand was a problem. High staff turnover and a lack of electronic tracking of standard information hindered program monitoring. A Mid-Atlantic Advisory Transportation Group reviewed the findings and made recommendations for service improvement. The Mid-Atlantic division evaluation contributed to an "evaluation synthesis" in which participants from the three divisions that had conducted Road to Recovery evaluations examined study data and made recommendations for reorganizing the national transportation program. A Transportation Program Design Team then held fact-finding meetings and adopted goals and objectives for a new national transportation program. The primary lesson learned was the far-reaching effects that a single program evaluation may have for various stakeholders and for an organization. PMID- 11912857 TI - The New England division Tell A Friend program implementation evaluation. AB - Tell A Friend (TAF) is the nationwide program of the American Cancer Society that aims to decrease breast cancer mortality by encouraging unscreened women to have mammograms using a peer-to-peer approach. An evaluation via the Collaborative Evaluation Fellows Project attempted to identify barriers and facilitators to successful implementation of TAF in the New England division. The proposed method included three surveys; this was revised because of a low response rate to the initial e-mailed survey. The actual method included one shortened survey and one follow-up survey. Results are presented for an initial and shortened survey for staff and a follow-up survey of respondents. Initial TAF implementation timelines were unrealistic, and the field staff encountered many barriers to implementation of TAF in the targeted communities. Changes are being made in training, management, and resources to ensure successful implementation. Continuous re evaluation is planned to ensure the successful implementation and conduct of the program. PMID- 11912858 TI - An external evaluator's perspective on the CEFP. AB - As the external evaluator of the Collaborative Evaluation Fellows Project (CEFP), the author describes how the CEFP model is applied to the program's own evaluation and presents early findings, mostly based on the 14 studies conducted in the first year of implementation. Outcomes, benefits, and design and implementation issues are examined. Early findings are that all of the basic premises of the CEFP model have proven feasible. The CEFP has been well received within the American Cancer Society and universities alike. The roles of the CEFP evaluation facilitator, evaluation faculty, and evaluation fellow have been assumed by appropriately qualified persons. The American Cancer Society staff and volunteers generally regard CEFP evaluations as trustworthy and useful, and, in most cases, they actually use them. Evaluation fellows place high value on the CEFP as a learning experience, and a surprising number pursue related careers. What was most feared at the CEFP planning stage--conflicts between the practical needs of the American Cancer Society and the academic orientations of its partners--have arisen much less than expected, probably because of careful provisions in the CEFP program design to guard against this. PMID- 11912859 TI - Lessons learned. What these seven studies teach us. AB - Seven evaluation studies of American Cancer Society programs offer findings useful for improving the programs studied and have larger implications for public health initiatives that rely on volunteers, the training of trainers, professional referral agents, and outreach through faith organizations or other indigenous community groups. Studies documented the following, for example: higher levels of productivity among volunteers who are cancer survivors, have health-related backgrounds, have college degrees, or both; the value of using existing organizations to recruit volunteers; the need for improved selection methods to reduce attrition in programs using "train the trainer" models; and the outreach potential of faith organizations as nodes in social networks within communities. The studies demonstrate to all voluntary health organizations the importance of assembling an accurate database to describe their activities and to enable them to answer important management questions. Lessons from these studies also can contribute to the development of better methods for the study of public health programs and for the incorporation of evaluation into the process of continuous program improvement within voluntary health organizations. PMID- 11912860 TI - Evaluation use and the CEFP. Lessons from a case study. AB - The Collaborative Evaluation Fellows Project was designed to build the evaluation capacity of the American Cancer Society and to produce evaluations that would be used in decision making on the local to national levels. Is this kind of use occurring and has use become a means of institutionalizing evaluation? Data from site visits to six regional offices suggest some use of findings occurring, while other data clearly indicate that an evaluation ethos and evaluation practice are both taking hold within the organization. PMID- 11912861 TI - The CEFP. Implications for public health education and training. AB - The 300-year history of the American academy outlines many of the contemporary pressures that are shaping the modern university. Faculty members need to balance the expectations of teaching, research, and service to amass a dossier that will lead to tenure. The academy needs to offer curricula that prepare graduates to enter the work force. Administrators need to encourage strong community ties to convince benefactors to invest in the renovation and expansion of university facilities. These pressures are especially acute in academic public health. The public health research agenda has extended from the study of infectious disease into behavioral risk and chronic disease. Schools of public health struggle to link curriculum, research, and service that will educate students, advance scholarship, and develop community interaction for the prevention of disease and the promotion of health. The Collaborative Evaluation Fellows Project provides a mechanism for the convergence and resolution of these pressures facing schools of public health. PMID- 11912862 TI - The CEFP. Building evaluation capacity. AB - The infusion of evaluation activity brought about by the American Cancer Society Collaborative Evaluation Fellows Project (CEFP) is a remarkable contribution to cancer prevention and control. Many lessons learned from the CEFP process are potentially beneficial not only to the American Cancer Society, but also to other organizations that seek to draw upon the success this project. One lesson for organizations that care about effectiveness and accountability is that, when outcomes matter, investments in evaluation become essential. By viewing evaluation in its larger context within the American Cancer Society, the CEFP delivers more value than the sum of findings from its individual studies. Especially pronounced are the contributions of the CEFP toward promoting organizational learning, generating useful products, and forming collaborative partnership--all of which are requirements for building an effective and sustainable evaluation system. PMID- 11912863 TI - Do environmental and hereditary factors affect the psychophysiology and left right shift in left-handers? AB - The principal objective of this study was to investigate the reality of sinistrality in left-handed subjects. The subjects were assessed by a 20-item questionnaire with two groups taken from Oldfield's and Yetkin questionnaires. The relation of different effects on left-hand preference was studied in men and women considering familial information and writing hand. The degree of hand preference was determined by Geshwind scores (GSs). The GS degrees of hand preferences were divided into weak and strong left-hand ranging from -20% to 100%. After assessing the GSs we asked the subjects to answer 22 questions with different aims, which were especially written for only left-handers. Some of them were based on familial sinistrality and hereditary relation to left-handers. The others concerned hand, foot, eye, shoulder, and ear preferences; the psychology of left-handedness; the knowledge concerned with left-handedness; the success, ability, and interest education and skill, and morphological differences in hand and foot sizes. In the present study, it was found that the subjects have had at least one left-handed person in their family or among their relatives. Some subjects had shifted their left hand preference in favor of their right hands. Nevertheless, the left-handers have been found as a presence with their own peculiarities. PMID- 11912864 TI - Evaluation of gingival recession in left- and right-handed adults. AB - Recession is the exposure of the root surface by an apical shift in the position of the gingiva. Several factors have been implicated in the etiology of gingival recession. The aim of this study was to assess the relationship between gingival recession and the hand, right or left, duration, frequency, and technique of tooth brushing in left- and right-handed adults in Erzurum, Turkey. Fifty five left-handed (35 female and 20 male) and 55 right-handed (35 female and 25 male) subjects were included in the study. The left and right hand positions of the subjects were determined by using Oldfield's Edinburg Inventory Index, and they were divided into two groups as left-handed and right-handed. The amount of gingival recession and localization, oral hygiene situations, gingival bleeding, toothbrushing duration, frequency, and techniques of subjects were evaluated. The relationship between the amount of gingival recession and localization and the subjects' hand, duration, frequency and technique of tooth brushing were examined. The data were evaluated with variance analysis and student's t test. The oral hygiene situations in right- and left-handed subjects were determined. On comparing the left-handed subjects with the right-handed ones, it was observed that the left-handed subjects had better oral hygiene than the right-handed. But, this case was not statistically significant (p > .05). In both right-handed and left-handed subjects, women had better oral hygiene than men (p < .01). The rate of gingival recession was found more in the left-handed than in the right-handed (p < .05). In the right-handed subjects, gingival recession was found in the premolar and canine regions of upper right and lower right jaw. A similar result was also observed in the left-handed subjects, because gingival recession was seen on their upper left and lower left jaw. Incisive teeth were affected at the same rate in the left-handed and right-handed. The gingival recession was seen more in maxillary jaw than in mandibular jaw in both groups. A statistically significant relationship between gingival recession and frequency duration, and technique of tooth brushing was found. While the greatest amount of gingival recession was found in horizontal scrub technique, gingival recession increased with increasing tooth brushing duration and frequency. The relationship between gingival recession and hand using in tooth brushing was determined. PMID- 11912865 TI - Neurobiological relationships between ambient lighting and the startle response to acoustic stress in humans. AB - A study was conducted to determine differences between psychophysiological responses to acoustic stress under different ambient lighting conditions. Typical cool-white indoor lighting was compared to full-spectrum lighting under dim and bright illumination levels. Full-spectrum lighting exhibits spectral characteristics that approximate those found in natural sunlight. The startle response was measured during three preliminary experiments using normal subjects. In the first experiment, one subject was evaluated. Electromyography (EMG) was used to measure eyeblink startle at the obicularis oculi muscle; heart rate (HR) was measured at the wrist; and galvanic skin conductance (GSC) was measured at the left index finger. There was a significant increase in galvanic skin conductance (GSC) and a decrease in heart rate. In the second experiment, GSC was used to measure the startle response under cool-white fluorescent lighting versus full-spectrum fluorescent lighting (n = 5 subjects) under both bright and dim lighting conditions. Higher arousal was detected measuring GSC under the cool white lighting condition, and under the dim lighting condition using both types of lighting. In the third experiment, ten subjects were assessed for GSC startle response using both lighting types set at the dim brightness level. There was a strong trend indicating a higher GSC arousal under the dim cool-white lighting condition. Other studies support that a GABAergic inhibitory circuit is inhibited by darkness and that the spectral characteristics of cool-white lighting are more stressful to humans than full-spectrum lighting. These preliminary findings regarding stress and ambient lighting may be especially important in the neural development of children, and in those clinical populations that are particularly sensitive to environmental stress. PMID- 11912866 TI - Neuropsychological profile of patients with primary systemic hypertension. AB - Arterial hypertension represents a risk factor for cerebrovascular disease. It has been hypothesized that chronic hypertension may eventually result in small subcortical infarcts associated with some cognitive impairments. One hundred fourteen patients with primary systemic hypertension (PSH) and 114 matched subjects were selected. PSH patients were further divided in four groups depending upon the hypertension severity. In addition to the medical and laboratory exams, a neuropsychological evaluation was administered. The NEUROPSI neuropsychological test battery was used. An association between level of hypertension and cognitive impairment was observed. Most significant differences were observed in the following domains: Reading, executive functioning, constructional, and memory-recall. No differences were observed in orientation, memory-recognition, and language. Some neuropsychological functions appeared impaired even in the PSH group with the least risk factors. Cognitive evaluation may be important in cases of PSH not only to determine early subtle cognitive changes, but also for follow-up purposes, and to assess the efficacy of different therapeutic procedures. PMID- 11912867 TI - Demographic effects on the Trail Making Test in marijuana abusers. AB - Demographic effects on the Trail Making Test (TMT), a test often used for screening for cognitive impairment, were examined in a sample of marijuana abusers in drug abuse treatment programs. A sample was drawn from electronic files of data from the Drug Abuse Treatment outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991-1993 in 96 programs in 11 cities in the United States. The number of marijuana abusers' scores available for analysis were 259. Data were analyzed to determine the effects of sex, ethnicity, age, and education variables on the two parts of the TMT in this large treatment sample of marijuana abusers. The variables of age and education level were statistically significantly related to TMT parts A and B, and ethnicity was statistically significant for part B of the TMT. R-Square values for overall models were moderate (A = .15, B = .18) suggesting that demographic effects on the TMT are weak. PMID- 11912868 TI - Demographic effects on the Trail Making Test in amphetamine abusers. AB - Demographic effects on the Trail Making Test (TMT), a test often used for screening for cognitive impairment, were examined in a sample of amphetamine abusers in drug abuse treatment programs. A sample was drawn from electronic files of data from the Drug Abuse Treatment Outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991-1993, in 96 programs in 11 cities in the United States. The number of amphetamine abusers scores available for analysis were 185. Data were analyzed to determine the effects of sex, ethnicity, age, and education variables on the two parts of the TMT in this large treatment sample of amphetamine abusers. No variables were statistically significant for either parts A and B of the TMT. R-square values for overall models were also negligible (A = .03, B = .08) suggesting that demographic effects on the TMT account for a minuscule amount of overall variance in terms of amphetamine abusers' TMT performance. PMID- 11912869 TI - Demographic effects on the Trail Making Test in sedatives abusers. AB - Demographic effects on the Trail Making test (TMT), a test often used to screen for cognitive impairment, were examined in a sample of sedatives abusers in drug abuse treatment programs. A sample was drawn from electronic files of data from the Drug Abuse Treatment Outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991-1993 in 96 programs in 11 cities in the United States. The number of sedatives abusers' scores available for analysis were 72. Data were analyzed to determine the effects of sex, ethnicity, age, and education variables on the two parts of the TMT in this large treatment sample of sedatives abusers. None of the demographic variables were statistically significantly related to TMT parts A and B, when considered as single variables. In addition, R-Square values for overall models were low (A = .15, B = .18), suggesting that demographic effects on the TMT in a sample of sedative abusers are very weak. PMID- 11912870 TI - Event-related synchronization and desynchronization during affective processing: emergence of valence-related time-dependent hemispheric asymmetries in theta and upper alpha band. AB - Event-related desynchronization (ERD) and synchronization (ERS) in the individually defined theta, alpha-1, alpha-2, and alpha-3 frequency bands were measured in 22 healthy subjects (Ss) in response to neutral (Neut), emotionally positive (Pos), and negative (Neg) IAPS stimuli. The 62-channel EEG, facial EMG and ECG were simultaneously recorded while subjects viewed sequentially presented pictures and subjectively rated them after each presentation. The obtained findings indicate that differences induced by pictures varying in emotional valence are associated mainly with increased theta and alpha-3 synchronization activity and anterior hemispheric asymmetries. In the anterior temporal leads theta ERS revealed a significant valence by hemisphere interaction showing relatively greater right hemisphere theta ERS for Neg and left hemisphere ERS for Pos stimuli in the time window of 100-700 ms post-stimulus, whereas in the alpha 3 band Neg stimuli induced lateralized time-dependent left hemisphere ERS increased in the time window of 800-1200 ms, were not observed for Neut and Pos stimuli. The obtained results along with earlier observations on EEG correlates of affective processing challenge the notion that affective anterior hemispheric asymmetries are mainly sensitive to wide alpha frequency band. Frequency and time dependence of anterior hemispheric asymmetries in emotional valence discrimination is emphasized. PMID- 11912872 TI - [Estrogen and progesterone receptors in meningiomas: immunohistochemical (Mib-1, p53) and clinico-morphological correlations]. AB - We studied 98 meningiomas including 89 benign, 5 atypical, and 4 anaplastic tumors to determine the immunohistochemical expression of estrogen and progesterone receptors and its prognostic value in comparison to histological grade, Mib-1 and p53. Estrogen and progesterone receptor positivity was observed in 63% and 5% of the cases, respectively. In 79% of meningiomas only a minimal proliferative activity was documented, whereas in 36% we detected an overexpression of the p53 oncoprotein. Anaplastic meningiomas were constantly negative for PgR, ER, and highly positive for Mib-1; 75% were positive for p53. A statistical correlation was demonstrated between p53 protein and Mib-1. Specifically the p53-negative meningiomas were frequently negative for Mib-1 (p = 0.002); conversely the lesions strongly positive for Mib-1 were p53-positive (p = 0.001). PMID- 11912873 TI - [Histopathological diagnosis of celiac disease in children and adults: 3 years' experience]. AB - We describe our personal experience with the routine histopathologic diagnosis of coeliac disease in pediatric and adult ages, over a period of 3 years. The most important indications based on the examination of over 300 cases include: (a) the importance of a correct orientation of biopsies on Millipore filters; (b) the not regular sending of clinical and laboratory results; (c) the better correspondence of modified Marsh classification by Oberhuber to the histologic characteristics of the disease before and after dieting; and (d) the increasing incidence of coeliac disease in adult age, even though in this case series the age-class most affected was that under 11 years (75 cases, 25%). PMID- 11912874 TI - [Significance of the AgNOR in tumor pathology]. AB - Analysis of silver-stained nucleolar organizer regions (AgNORs), originally regarded as a diagnostic tool, is now considered mainly as a prognostic parameter. Indeed, the expression of AgNOR proteins is associated with several biological properties of neoplastic cells: metabolic activity, DNA content, histological grade of differentiation and, especially, the rapidity of cellular proliferation. Thus, a high AgNOR quantity is a marker of aggressive tumour phenotype, and a large number of papers have shown the independent prognostic value of AgNOR analysis in several human neoplasias. Moreover, the method can be applied to small biopsies, can identify neoplastic clones with different proliferative activities and may stratify patients into different risk groups. The standardized method for AgNOR quantification offers objective and reproducible results. The evaluation of AgNOR quantity in cycling cells, either by immunohistochemistry or by a novel flow cytometry technique, may represent the future of AgNOR analysis. PMID- 11912875 TI - Biological aspects of a Brazilian strain of Entamoeba dispar. AB - A strain of Entamoeba dispar was characterized by clinical diagnosis, serological and electrophoretical isoenzyme analysis and by the polymorphism of a 482 bp genomic fragment analysis. The pathogenesis and virulence of this strain was investigated considering the experimental infection in hamster livers in association with the original intestinal microbiota. Liver lesions were observed in hamsters experimentally infected with trophozoites from xenic cultures, but not from the monoxenic cultures. Moreover, clones obtained from re-isolated strain Wil1R1 showed a distinct biological behavior. In fact, animals inoculated with Wil1R1ClB3 showed an intense acute inflammatory reaction with destructive focal hepatic lesions. These lesions were characterized as amebic abscesses. The association between bacteria and ameba has been fairly well studied because it affects the pathogenicity of the amebas and has important therapeutic implications. In this study, we demonstrated that E. dispar in association with the original microbiota is able to produce lesions in hamster liver in spite of its having been considered to be non-pathogenic in the hamster model. Based on these results we suggest that diagnosis of amebiasis needs to be made with more care and that clinical and therapeutical procedures need to be revised. PMID- 11912876 TI - PCR detection of specific Leishmania-DNA in patients with periodontal disease. AB - This study deals with the detection of Leishmania braziliensis DNA in gingival specimens from 10 individuals who all had suffered from cutaneous leishmaniasis 5 10 years prior to the examination and all had been treated with anti leishmaniasis drugs. This preliminary study gives an interesting contribution to the oral microbiology of this disease, with the observation that inflamed periodontal tissues can serve as a factor affecting the dispersion of Leishmania parasites in individuals who had suffered from cutaneous leishmaniasis. These finding are corroborated by the results of polymerase chain reaction (PCR) which demonstrated the presence of Leishmania DNA in tissue samples of patients with periodontal diseases. PMID- 11912878 TI - [The clinico-pathological conference as an instrument for improving diagnostic performance ]. PMID- 11912877 TI - [Pulmonary carcinoma with myoepithelial differentiation, analogous to basal cell adenocarcionma of the salivary glands: description of a case]. AB - We describe a case of pulmonary carcinoma with myoepithelial differentiation, analogous to basal cell adenocarcinoma of salivary glands. The patient, a 60 year old man, smoker, presented with three peripheral nodules of the left lung. Preoperative staging was negative for metastatic disease and the patient underwent a surgical resection of the nodules. After 22 months, the patient is alive with no evidence of disease. Microscopically, the tumours were composed of atypical cells arranged in lobules, separated by basal membrane-like material. Immunohistochemically, tumour cells were positive for cytokeratin AE1/AE3, cytokeratin 14, vimentin, calponin, S-100 protein and gliofibrillary acid protein (GFAP). Electron microscopy showed features of epithelial and myoid differentiation and confirmed the myoepithelial nature of the tumour. Pulmonary tumours with myoepithelial differentiation are rare, but they have a wide and distinctive morphological spectrum. PMID- 11912880 TI - [From the Italian Gynecologic Pathology Society, a proposal for gynecologists and pathologists: to study congenital neurological pathology in Italy]. PMID- 11912879 TI - [Robert Koch (1843-1910): a life in the trenches (against microbes, against VIrchow, and against so many other things)---and an unexpected parallel with Golgi]. PMID- 11912881 TI - [New TNM classification for melanoma: final version]. PMID- 11912882 TI - [Document from the Italian Group for Pathological Anatomy of the Embryo, Fetus, and their Appendages and the Italian Cardiovascular Pathology Study Group ]. PMID- 11912883 TI - [The Thin Prep Pap Test; a platform for gynecologic diagnosis]. PMID- 11912884 TI - Reports say diversion on the rise: use technology to overhaul patient flow. AB - Your patient flow can be dramatically improved with electronic patient tracking systems that give information in real time. If ED staff can spot delays at a glance, steps can be taken to correct the root of the problem before backlog occurs. If specific time goals aren't met, an immediate intervention should occur. An "ED dashboard" system gives immediate information about lab results, which can reduce overall length of stay significantly. PMID- 11912886 TI - ED's disaster plan uses incident command system. PMID- 11912885 TI - Improve care of kids with pediatric coordinator. AB - New guidelines from the American Academy of Pediatrics and the American College of Emergency Physicians recommend having physician and nurse pediatric coordinators to improve the care of children in the ED. The role of pediatric coordinator doesn't need to be a full-time or newly added position. Physician and nurse coordinators should work together to develop policies for improved care. Coordinators should routinely audit patient charts as part of a quality improvement program. PMID- 11912887 TI - Anthrax aftermath: adverse drug reactions, vaccine controversy undercut CDC extended treatment offer. PMID- 11912888 TI - Web site designed to give clinicians source to get info on near misses. PMID- 11912889 TI - Improving patient flow: the Esther Project. PMID- 11912890 TI - Press, Ganey winners are best in customer care. PMID- 11912891 TI - Tracking cost and info: an affordable solution. PMID- 11912893 TI - Selection & use of telehealth technology in support of homebound caregivers of stroke patients. PMID- 11912892 TI - Improving the quality of home care for patients with heart failure. AB - Home care nurses play a very important role in caring for patients with heart failure. The home care nurse's ability to follow patients one-on-one over a period of time, assess and observe patient self-care behaviors such as medication adherence and dietary intake, and intervene with patient education is unique. PMID- 11912894 TI - Strength in unity. AB - Mary Suther is a legend in home care. She has been in home care for many, many, many years, long before there was the National Association for Home Care. In fact, she was instrumental in the creation of the National Association for Home Care. NAHC has grown and prospered in large part to the leadership of Mary Suther, who has been on the Board for years in a variety of different roles and is our Chairman. She is a truly great individual. Mary Suther gave the following speech at NAHC's 2001 Annual Meeting. We reprint it here for the benefit of our readers. PMID- 11912895 TI - Looking for clues. A discussion of cardiac event recorders in home healthcare practice. AB - Home healthcare nurses have a unique opportunity to observe patients closely as they go about their activities of daily living within their own environment. But nurses must be alert to assess for all signs and symptoms and not ignore those that do not relate solely to the primary diagnosis care. PMID- 11912896 TI - Post-surgical cardiac patients receive new level of care. AB - VNA Home Health Systems in Orange County, California provides intermittent home care for homebound and critically ill patients. The VNA administration recognized the advantages of clinicians using a pocket-sized cardiac monitor. Eager to continually adopt technologies that improve their level of care, the pocket-sized cardiac monitor was a logical addition to VNA's PDA-based clinical tools. PMID- 11912897 TI - Why offer home cardiac recovery care? AB - Why offer cardiac recovery specialty programs in home care? The answer to this question lies in the issue related to nursing professionals' obligation to provide best practice for all patients. Home cardiac care that complies with best practice, evidence-based guidelines, derives the best patient outcomes. A comprehensive and best-practice approach to care is simply what every patient recovering from a cardiac event deserves. PMID- 11912898 TI - Ambulatory electrocardiography in home care. PMID- 11912899 TI - Reversible conversion of nitroxyl anion to nitric oxide. PMID- 11912900 TI - Purification and determination of activity of mitochondrial cyanide-sensitive superoxide dismutase in rat tissue extract. PMID- 11912901 TI - Studies of metal-binding properties of Cu,Zn superoxide dismutase by isothermal titration calorimetry. PMID- 11912902 TI - Superoxide reductase from Desulfoarculus baarsii. PMID- 11912903 TI - Enzyme-linked immunosorbent assay for human and rat manganese superoxide dismutases. PMID- 11912904 TI - Investigating phenotypes resulting from a lack of superoxide dismutase in bacterial null mutants. PMID- 11912905 TI - Bacterial superoxide dismutase and virulence. PMID- 11912906 TI - Superoxide dismutase null mutants of baker's yeast, Saccharomyces cerevisiae. PMID- 11912907 TI - Measurement of "free" or electron paramagnetic resonance-detectable iron in whole yeast cells as indicator of superoxide stress. PMID- 11912908 TI - Transgenic superoxide dismutase overproducer: murine. PMID- 11912909 TI - Transgenic and mutant mice for oxygen free radical studies. PMID- 11912910 TI - Overexpression of Cu,ZnSOD and MnSOD in transgenic Drosophila. PMID- 11912911 TI - Manganese porphyrins and related compounds as mimics of superoxide dismutase. PMID- 11912913 TI - Superoxide dismutase mimics: antioxidative and adverse effects. PMID- 11912912 TI - Reactions of manganese porphyrins and manganese-superoxide dismutase with peroxynitrite. PMID- 11912914 TI - Superoxide reductase activities of neelaredoxin and desulfoferrodoxin metalloproteins. AB - Superoxide reductases have now been well characterized from several organisms. Unique biochemical features include the ability of the reduced enzyme to react with O2- but not dioxygen (reduced SORs are stable in an aerobic atmosphere for hours). Future biochemical assays that measure the reaction of SOR with O2- should take into account the difficulties of assaying O2- directly and the myriad of redox reactions that can take place between components in the assay, for example, direct electron transfer between cytochrome c and Dfx. Future prospects include further delineation of the reaction mechanisms, characterization of the putative (hydro)peroxo intermediate, and studies that uncover the components between reduced pyridine nucleotides and SOR in the metabolic pathway responsible for O2- detoxification. PMID- 11912915 TI - Purification and preparation of prion protein: synaptic superoxide dismutase. PMID- 11912916 TI - Mitochondrial superoxide anion production and release into intermembrane space. AB - The topological distribution of ubiquinone in the mitochondrial respiratory chain suggests that both ubiquinone pools may participate in O2.- production and, hence, are vectorially released into the matrix and intermembrane space. Mitoplasts, obtained by either digitonin or hypotonic KCl treatment, are a suitable experimental model for measuring O2.- in the intermembrane space. The use of membrane-impermeable spin-broadening agents strengthens the notion that part of the O2.- generated by the respiratory chain may be released into the intermembrane space. This, together with the putative occurrence of a Cu, Zn superoxide dismutase in this compartment may account for part of H2O2 released by mitochondria and contributing to a cytosolic steady-state level of this species in cytosol. PMID- 11912917 TI - Measurement of superoxide radical and hydrogen peroxide production in isolated cells and subcellular organelles. PMID- 11912918 TI - Biochemical assay of superoxide dismutase activity in Drosophila. PMID- 11912919 TI - Transcriptional regulation and environmental induction of gene encoding copper- and zinc-containing superoxide dismutase. PMID- 11912920 TI - Quantitation of intracellular free iron by electron paramagnetic resonance spectroscopy. PMID- 11912921 TI - Transcription regulation of human manganese superoxide dismutase gene. AB - The human MnSOD gene has a typical housekeeping gene promoter, but is highly inducible by various physical, chemical, and biological agents. Transcription factors SP-1 and AP-2 seem to have opposite roles in the transcriptional activity of the basal promoter. Whereas SP-1 plays a positive role, which is absolutely essential for transcription from the human MnSOD promoter, AP-2 appears to play a negative role in this process. An enhancer element is found in the promoter region of the human MnSOD gene. Several important enhancer elements are located in the second intron. The NF-kappa B site in the second intron is essential but not sufficient for high-level induction of MnSOD by cytokines. Although mutations in the regulatory elements may be partially responsible for the lack of induction of MnSOD in some cell types, differences in the degree of induction exist that cannot be accounted for by the defect in the DNA sequence. It is highly likely that this difference is due to the presence or absence of coactivator or suppressor proteins in the cells and may have a physiological role in the defense against oxidative stress. PMID- 11912922 TI - Assessment of oxidants in mitogen-activated protein kinase activation. PMID- 11912923 TI - Assaying binding capacity of Cu,ZnSOD and MnSOD: demonstration of their localization in cells and tissues. PMID- 11912924 TI - Superoxide dismutase in aging and disease: an overview. PMID- 11912925 TI - Tissue-specific mitochondrial production of H2O2: its dependence on substrates and sensitivity to inhibitors. PMID- 11912926 TI - Targeting superoxide dismutase to critical sites of action. PMID- 11912927 TI - In vitro quantitation of biological superoxide and hydrogen peroxide generation. PMID- 11912928 TI - Superoxide dismutase kinetics. PMID- 11912929 TI - Analysis of Cu,ZnSOD conformational stability by differential scanning calorimetry. PMID- 11912930 TI - Catalytic pathway of manganese superoxide dismutase by direct observation of superoxide. AB - Measurement of catalysis by MnSOD using direct observation of the UV absorbance of superoxide allows determination of steady-state catalytic constants. Stabilizing superoxide in aprotic solvents such as dimethyl sulfoxide permits the use of stopped-flow spectrophotometry, although significant information is lost in the 2- to 4-msec mixing time; generating superoxide by pulse radiolysis requires no mixing time. Studies show that kcat/Km for the decay of superoxide catalyzed by MnSOD proceeds at diffusion control. Investigations using solvent hydrogen isotope effects and enhancement of catalysis by exogenous proton donors show that kcat near 10(4) sec-1 contains a significant contribution from proton transfer steps. The active site of MnSOD is dominated by a hydrogen bond network comprising the manganese-bound aqueous ligand, the side chains of four residues (Gln-143, Tyr-34, His-30, and Tyr-166 from an adjacent subunit), as well as other water molecules. Interrupting this hydrogen bond network by conservative replacement of residues 30, 34, and 166 causes a 10- to 40-fold decrease in maximal velocity, interpreted as an effect on proton transport to the active site, with smaller effects on kcat/Km. Replacement of Gln-143 causes a much greater decrease in catalytic activity, by two to three orders of magnitude, and causes significant changes to the redox potential as well. During catalysis, MnSOD is inhibited by a peroxide complex of the metal in the active site, different from the inhibition of FeSOD and Cu,ZnSOD by Fenton chemistry. Site specific mutagenesis of active-site residues alters the extent of product inhibition of MnSOD as well, indicating that this is not only a property of the metal. The replacement of Trp-161 with phenylalanine results in a variant that is completely blocked in catalysis by product inhibition. PMID- 11912931 TI - Extracellular superoxide dismutase. PMID- 11912932 TI - Prokaryotic manganese superoxide dismutases. PMID- 11912933 TI - Aconitase: sensitive target and measure of superoxide. PMID- 11912934 TI - Nickel-containing superoxide dismutase. PMID- 11912935 TI - Psychiatric presentations of non-HIV infectious diseases. Neurocysticercosis, Lyme disease, and pediatric autoimmune neuropsychiatric disorder associated with streptococcal infection. AB - Infectious diseases can cause an array of symptoms, including psychiatric symptoms. Psychiatrists serving the medically ill need to be aware not only of classic infectious diseases (e.g., neurosyphilis and HIV), but also of less commonly discussed infectious diseases (e.g., NCC, PANDAS, and Lyme disease). These examples represent an internationally endemic disease (e.g., NCC), a probable immunogenetic disease (e.g., PANDAS), and a frequently overdiagnosed and overtreated disease (Lyme disease). PMID- 11912937 TI - Practical psychopharmacology in HIV-1 and acquired immunodeficiency syndrome. AB - HIV-1 infection poses a challenge for psychiatrists of the medically ill. Many factors concerning the care of HIV-1-infected patients need to be considered when prescribing psychotropics. These include careful diagnosis, taking into account medical disorders associated with HIV-1 that can present with psychiatric symptoms, as well as medications that HIV-1 patients may be taking that can cause a variety of neuropsychiatric side effects. Another important issue is the potential for drug-illness interactions. In general, HIV-1 patients seem to be more sensitive to the development of adverse drug reactions than do non-HIV-1 patients, especially as the illness progresses. It is also important to be cognizant of the complex multidrug regimens that many HIV-1 patients are on to avoid known drug-drug interactions and be on the alert for other potential interactions when using psychotropic medications. PMID- 11912938 TI - The use of electroencephalography in psychiatry of the medically ill. AB - The psychiatrist considering recommending an EEG should look for acute changes in the history or examination suggestive of an organic cause. If he or she judges that the EEG will help to clarify or confirm the diagnostic impression already formulated, it is worth considering whether adding provocative maneuvers could increase the yield. The authors cannot overemphasize the importance of using the EEG in correlation to further inform old-fashioned clinical detective work already in process, particularly when the EEG could rule out a potential organic contributor to a psychiatric phenotype. For routine screening without an elevated index of suspicion or for thoughtless "fishing expeditions," EEG results will surely disappoint. PMID- 11912936 TI - Psychiatry of the medically ill in the burn unit. AB - Clinical experience and burn survivor testimony show that the experience of being burned can be associated with catastrophic stress and lead to drastic permanent body image changes from scarring and limb-function loss. Close relatives, if not killed in the fire, often also experience clinically significant bystander stress. Closeness of relationships may be lost, and self-image may suffer. Property damage and loss of crucial resources may be associated with fires. Although many burns result from accidents, most result from preventable causes associated with psychiatric disorders, which include mood disorders, psychoses, cognitive disorders, and substance-use disorders. Burns then result from: Deliberate self-harm Impaired judgment and poor coordination associated with substance intoxication Risk-taking behavior Poor supervision of children and impaired elderly persons Careless handling of flammable materials. Many clinical syndromes, such as delirium, ASD, acute psychosis, suicidality, and pain need to be addressed by the consulting psychiatrist to facilitate surgical treatment of the burn injury. Other psychiatric disorders, such as PTSD, major depression, and adjustment disorder, need to be treated to expedite long-term adjustment. Hospital length of stay and RTW/RTS are major outcome variables. The psychiatry consultant can positively affect both variables substantially using both pharmacologic and psychosocial measures. The important role of psychiatric issues both before and after burn injury support the need for more consistent and comprehensive medical insurance coverage for psychiatric consultation to burn units and clinics. Burn Support Groups are an invaluable asset. PMID- 11912939 TI - Electroconvulsive therapy in the medically ill. AB - ECT is often a necessary treatment for severe psychiatric disorders in patients with medical or neurologic comorbidity. Although the available data consist largely of cases and case series, ECT is effective in treating psychopathology despite the comorbidity. With appropriate precautions and monitoring during and after ECT, complications can be minimized. PMID- 11912940 TI - Liver transplantation in patients with alcohol and other substance use disorders. AB - It is unfortunate that LT is not the ultimate sobering experience. LT patients can and do relapse; however, relapse to alcohol or substance use should no more be considered a failure of LT than the recurrence of HCV after LT. It is a phenomenon of their addiction. As a group, the survival and outcomes of patients undergoing LT for ALD are not significantly different from those undergoing LT for other causes. The fact that few patients return to heavy and deleterious alcohol or substance use attests to the success of programs in selecting patients capable of caring for themselves and their livers after LT. In the pre-LT phase, establishing the correct addiction diagnosis is essential so that those at highest risk can be carefully monitored. Although information is emerging about other risk factors for relapse, the authors caution against considering patients with these characteristics as being categorically unfit for LT. Each individual is unique, and such factors should guide clinical decision making rather than being absolute contraindications. After LT, clinical interviews, preferably by a trained psychiatric physician or clinician, are essential to monitoring alcohol and substance use, and surveillance early on is required. Much work needs to be done with respect to substance use and relapse after LT because few studies have explored these issues. In addition, the special treatment needs of those who relapse has not been addressed. Treatment research is underway but has relied on traditional strategies that are not always applicable to LT recipients. Further areas to improve clinical care include improving health behaviors, specifically, smoking cessation. The authors anticipate that, in the near future, the ongoing work in this area will provide information and guidance to physicians and clinicians caring for these unique patients. PMID- 11912941 TI - Herbal and nonherbal supplements in medical-psychiatric patient populations. AB - Nutritional supplements remain a popular choice for patients seeking relief or prevention from a wide range of physical and mental disorders. Review of available literature finds support for some therapeutic uses, but flaws in study design (e.g., small sample sizes) and methodology (e.g., inadequate blinding and a lack of placebo control) limit the possibility of making strong recommendations. Quality-control issues also raise concerns about the safety of supplement use (see previous list). In addition, potential interactions with prescription drugs are another consideration (Table 1). By becoming more knowledgeable about the risks and benefits of nutritional supplements, psychiatrists can assist patients in making informed choices and avoiding unnecessary harm. PMID- 11912942 TI - Designer drugs in the general hospital. AB - This article has reviewed the potential complications of acute intoxication and withdrawal from some of the more commonly used club, or designer, drugs. Although limited, acute use of these drugs is claimed by users to be benign, in the context of crowded raves and circuit parties, where multiple drugs may be used, hyperthermia, dehydration, and life-threatening reactions may occur. In addition, mounting evidence of the long-term effects of continued use of these drugs is cause for great concern. Finally, awareness of a severe withdrawal syndrome from GHB and its precursors is particularly important to psychiatrists of the medically ill, who may be called on to help in the management of these patients. PMID- 11912943 TI - Psychiatric consequences of motor vehicle accidents. AB - Psychiatric complaints are frequent following motor vehicle accidents and may be major predictors of persistent pain and other complaints. Outcomes are not related closely to the nature or severity of any medical injury. Psychiatric problems often are unrecognized and untreated. There is a need for more behaviorally inferred routine care, early recognition of complications, and the use of psychological and pharmacological interventions. PMID- 11912944 TI - Psychiatric aspects of traumatic brain injury. AB - TBI is a complex heterogenous disease that can produce a variety of psychiatric disturbances, ranging from subtle deficits in cognition, mood, and behavior to severe disturbances that cause impairment in social, occupational, and interpersonal functioning. With improvement and sophistication in acute trauma care, a number of individuals are able to survive the trauma but are left with several psychiatric sequelae. It is important for psychiatrists to be aware of this entity because an increasing number of psychiatrists will be involved in the care of these patients. Treatment should be interdisciplinary and multifaceted, with the psychiatrist working in collaboration with the patient, caregiver, family, other physicians, and therapists. The goal of treatment should be to stabilize symptoms; maximize potential; minimize disability; and increase productivity socially, occupationally, and interpersonally. PMID- 11912945 TI - Refractory chronic pain. AB - Refractory chronic pain is a significant public health problem and frustrating to everyone affected by it. All physicians can participate in the care of these patients, but psychiatrists should take a leading role in their care. A comprehensive approach offers hope and increases the opportunities for successful treatment. Each perspective of an interdisciplinary formulation has a unique logic that defines specific methods for designing treatment for the patient with refractory chronic pain. The patient does not have to fit into one theoretic approach or model to receive and accept treatment. The patient's diagnoses are based on the formulation, which then directs treatment along rational directions. The linkages and interactions of a patient's diagnoses can then be investigated within a framework that includes the entire person and not just his or her biochemistry. If a patient's suffering persists, other factors must be considered that may have been overlooked before the treatment plan is abandoned or modified. Usually these factors are within one of the perspectives initially thought to be less important. A new combination of approaches is then required to treat the patient successfully. The perspectives appreciate not only that the patient is struggling through important life events but also that he is a person composed of vulnerabilities and strengths, having made many choices and afflicted by diseases. PMID- 11912946 TI - Psychiatric issues in pulmonary disease. AB - This article has attempted to provide an overview of the clinical literature regarding the psychological issues facing patients with pulmonary disease, depending on when the illness begins in the life span, because different developmental tasks are disrupted. Patients must contend with side effects of medication that may mimic or exacerbate psychiatric disorders. The main drug interactions for psychiatrists to be aware of in this patient population occur between rifampin, or theophylline and psychotropic medications. In lung transplant recipients on cyclosporine therapy, the antidepressant drug nefazadone may cause increased cyclosporine levels. Psychiatrists must be aware of the risks, benefits, and survival statistics; educate patients; and ascertain whether the patient is competent to make medical decisions regarding treatment procedures. PMID- 11912947 TI - [Influence of diabetes mellitus on the initial and long-term outcome of patients treated with coronary stenting]. AB - OBJECTIVES: The long-term outcome of patients with diabetes who underwent stent placement has not been assessed comprehensively. METHODS: Analysis of angioplasty procedures performed between January 1994 and December 1998 identified 140 diabetics (156 lesions) and 169 non-diabetics (187 lesions) who underwent successful stent placement. Follow-up was completed in 286 patients (93%) with a mean follow-up period of 2.8 +/- 1.3 years. Cardiac death, nonfatal myocardial infarction, coronary artery bypass grafting and repeat percutaneous transluminal coronary angioplasty were considered as adverse cardiac events. The primary clinical endpoint was event-free survival at 1 and 3 years. The primary angiographic endpoint was restenosis rate at follow-up angiography (follow-up rate 75% of 257 lesions). RESULTS: The diabetics were older (66 +/- 8 vs 62 +/- 11 years, p < 0.0005) with more risk factors such as hypertension (69% vs 57%, p < 0.05) and multivessel disease (69% vs 51%, p < 0.005). Final balloon size was smaller in diabetics than in non-diabetics (3.26 +/- 0.61 vs 3.39 +/- 0.53 mm, p < 0.05). Restenosis rate was significantly higher in diabetics than in non diabetics (36% vs 24%, p < 0.05), but the target lesion revascularization in diabetics was not statistically different compared with non-diabetics (22% vs 16%). Long-term event-free survival was not significantly different between diabetics and non-diabetics (69.9% vs 74.8% at 1 year, 57.3% vs 66.0% at 3 years). CONCLUSIONS: Diabetics have an increased risk for angiographical restenosis after successful stent placement compared to non-diabetics. However, diabetics who underwent stent placement had a favorable clinical long-term outcome similar to non-diabetics. PMID- 11912948 TI - [Assessment of left ventricular function by midwall fractional shortening in hemodialysis patients]. AB - OBJECTIVES: Midwall fractional shortening (MFS) is a useful index to evaluate left ventricular myocardial function in patients with essential hypertension. The study investigated the prevalence and characterization of low MFS in hemodialysis patients. METHODS: MFS was calculated from M-mode echocardiograms in 67 patients (34 males, 33 females) receiving maintenance hemodialysis in whom fractional shortening was normal. Plasma levels of atrial and brain natriuretic peptides were also measured in these patients before and after hemodialysis. MFS was evaluated by stress-corrected MFS (ratio of observed to predicted MFS). The relationship of MFS to circumferential end-systolic stress in 122 healthy subjects was used to calculate the predicted MFS. RESULTS: Stress-corrected MFS was depressed in 18 of the 67 patients (26.9%). In the low MFS group, duration of hypertension was significantly longer (p < 0.05), wall thickness was significantly greater (p < 0.001), left ventricular dimension was significantly smaller (p < 0.0001), and relative wall thickness was significantly greater (p < 0.0001) than in the normal MFS group. Reduction of brain natriuretic peptide level by hemodialysis in the low MFS group was significantly higher (p < 0.05) than in the normal MFS group. CONCLUSIONS: Depression of stress-corrected MFS may be common in hemodialysis patients. Long duration of hypertension and concentric geometry of the left ventricle occur in patients with low MFS. PMID- 11912949 TI - Serum calcium concentration and carotid artery plaque: a population-based study. AB - OBJECTIVES: Serum calcium level may be associated with the morbidity and mortality of cerebrovascular and cardiovascular diseases. However, only a few large-scale population studies have been performed to investigate the association between serum calcium and carotid plaque. METHODS: A retrospective cross sectional study was performed on the subjects who underwent general health screening tests including ultrasonographic evaluation of the carotid artery between 1994 and 2000 at our institute. Before the statistical analysis, all serum calcium values were adjusted for the serum albumin concentration. RESULTS: Of 5,732 subjects enrolled in the present study, 3,785 were male and 1,947 were female, and were aged 22-88 years (median 57 years). Carotid plaque was identified in 1,313 (23%) subjects. Serum calcium concentration was slightly greater in the subjects with plaque than in those without (2.28 +/- 0.8 vs 2.27 +/- 0.7 mmol/l, p < 0.001; unpaired t-test). Multivariate logistic regression analysis including confounding risk factors revealed that serum calcium is an independent positive predictor for carotid plaque with an odds ratio of 1.70 [95% confidence interval (CI): 1.50-1.92] for each increase of 1 mg/dl. Male and female subjects in the highest quartiles of serum calcium concentrations had a greater risk of carotid plaque with odds ratios of 1.52 (95% CI 1.35-1.71, p < 0.01) and 1.57 (95% CI 1.27-1.92, p < 0.05), respectively, compared to the subjects in the lowest quartiles of calcium concentrations. CONCLUSIONS: These findings indicate that serum calcium is an independent risk factor for carotid plaque. PMID- 11912950 TI - Antiarrhythmic effect of nifekalant on atrial tachyarrhythmia in four patients with severe heart failure. AB - OBJECTIVES: Nifekalant is a class III antiarrhythmic drug, which prolongs the refractory period of the atrial and ventricular myocardium, without negative inotropic action. Intravenous nifekalant was administered in four patients with atrial tachyarrhythmia and severe heart failure to terminate or prevent atrial tachyarrhythmia. METHODS AND RESULTS: Two of three episodes of atrial tachyarrhythmia were terminated by intravenous nifekalant (0.22 to 0.30 mg/kg) administration. Continuous intravenous infusion of nifekalant (0.15 to 0.40 mg/kg/hr) during six episodes to maintain the sinus rhythm, prevented recurrence of atrial tachyarrhythmia in five episodes in which prolongation of the QTc interval was observed to more than 450 msec. None of the patients showed worsening of the hemodynamics during treatment. One patient developed polymorphic ventricular tachycardia, which deteriorated into ventricular fibrillation. CONCLUSIONS: Nifekalant may be effective for treating atrial tachyarrhythmia in patients with severe heart failure. Further clinical studies are needed to confirm these findings. PMID- 11912951 TI - [Efficacy of biventricular pacing on myocardial glucose metabolism in a patient with heart failure using 2-deoxy-2[18F]fluoro-d-glucose positron emission tomography: a case report]. AB - A 61-year-old-male with dilated cardiomyopathy and congestive heart failure was treated by implantation of a biventricular pacemaker. Myocardial glucose metabolism was evaluated by positron emission tomography with 2-deoxy 2[18F]fluoro-d-glucose (FDG) during atrial-right ventricular pacing or atrial biventricular pacing. Percentage uptake of FDG decreased in the lower septal area of the left ventricle during right ventricular pacing, but the decrease was improved during biventricular pacing (74.7% vs 91.6%). Biventricular pacing may improve myocardial glucose metabolism in patients with congestive heart failure. PMID- 11912952 TI - [Infective endocarditis complicating bilateral bacterial ophthalmitis: a case report]. AB - A 53-year-old female suddenly went blind in her left eye on 3 June, 2000. She was admitted to the Department of Ophthalmology of our hospital under the diagnosis of endophthalmitis. Her left eye was enucleated, and Streptococcus agalactiae was found in the vitreous fluid. After left ophthalmectomy, inflammation recurred after cessation of antibiotic administration. Echocardiography demonstrated a vegetation of the posterior mitral valve. The diagnosis was infective endocarditis. She was transferred to the Department of Internal Medicine. Mitral regurgitation deteriorated during the course of medical therapy, but she was discharged on 13 September, 2000 because inflammation had improved remarkably and the vegetation had disappeared after administration of penicillin G, panipenem, cefotaxime and clindamycin. We suspected that embolism of the ophthalmic artery was the cause of the sudden blindness in her left eye. Infective endocarditis with bacterial endophthalmitis is very rare in Japan. PMID- 11912953 TI - [Acute inferior myocardial infarction with marked ST elevation in leads V1 to V3 after coronary artery stenting]. PMID- 11912954 TI - [Aortic valve tumor detected by two-dimensional echocardiography in a patient after myocardial infarction]. PMID- 11912955 TI - Team approach saves money. PMID- 11912956 TI - Protect outpatient revenue. PMID- 11912957 TI - Just say no to fixed payments. PMID- 11912959 TI - Policy. Health care hits a speed bump. PMID- 11912958 TI - A continuum of care for elderly. PMID- 11912960 TI - Finance. Patience required. PMID- 11912961 TI - Facility renovation. Free labor. PMID- 11912962 TI - Public hospitals. A welcome mat for the uninsured. PMID- 11912963 TI - Energy management. A bright idea. PMID- 11912964 TI - 1Q[3a]. Will health reform happen at the state or federal level?. PMID- 11912965 TI - Cardiac care. Lab challenged. PMID- 11912966 TI - Long-term care. The battle against bacteria and fear. PMID- 11912967 TI - The smallest table. PMID- 11912974 TI - Thinking ahead. Three hospitals demonstrate that you can plan for the future, even if you can't forsee it. AB - Predicting the future is next to impossible, but that doesn't grant health care administrators a pass at planning for it. How do hospitals build new facilities to anticipate trends that they haven't heard of yet? Here's a look at three hospitals--a rural facility, a new set of operating rooms and a new ED--that are taking the risk. PMID- 11912975 TI - Leveraging the cost of HIPPA.. AB - HIPAA's administrative simplification rule was introduced to reduce the costs of handling provider-payer transactions by standardizing them. The transaction rules are predicted to save billions of dollars. To get the biggest payoff, providers will need to overhaul their business processes as part of electronic transaction processing. By eliminating redundant and inefficient administrative processes, staff time can be focused on processes that improve financial performance and customer satisfaction. If your organization needs to have an extension beyond the original compliance date, you must file a plan before Oct. 16 to show how you'll come into compliance by Oct. 16, 2003. Check the roadmap and timetable for HIPAA compliance and see where your organization is and where it needs to be. PMID- 11912976 TI - It's not easy. But collaboration is the best--maybe only--way to improve your community's health. PMID- 11912977 TI - Property insurance. Hospital rates go through the roof. PMID- 11912978 TI - Executive turnover. Hello, CEO, goodbye. PMID- 11912979 TI - Regulations. The simpler life. PMID- 11912980 TI - For-profit health care. Hospital stocks prove health on Wall Street. PMID- 11912982 TI - E-commerce. Treatment on the Net. PMID- 11912981 TI - Benefits dilemma. PMID- 11912983 TI - 1Q[3a]. What can be done to increase organ donation. PMID- 11912984 TI - Patient safety. Safe. Sound? PMID- 11912985 TI - Quality care. Measuring up. PMID- 11912986 TI - Winning isn't everything. PMID- 11912987 TI - 2002 Benchmarking Guide. AB - The profitability gap between hospitals in high and low managed care markets is closing. The difference in total margins is down and the change in return on equity is even more dramatic. However, even with heavier patient care demands, hospitals in high managed care markets have a firmer grip on expenses and show productivity gains. Our annual Benchmarking Guide shows that the number of HMOs operating in the United States continues to decline with industry mergers, plan consolidations and plan closures. While 2001 enrollment stabilized at 79.5 million, the number of HMOs fell to 541. The guide also compares data on hospital owned and non-hospital-owned physician practices. PMID- 11912988 TI - Prescription for concern. AB - As U.S. medical care relies more heavily on prescription drugs, hospitals are caught in an increasingly painful situation. Shortages of critical pharmaceuticals often leave hospitals empty-handed and, according to clinicians, endanger patient safety. Soaring drug costs account for a huge proportion of burgeoning health care spending, and strategies to control costs, including pharmacy benefit managers and drug discount cards for seniors, so far have had limited or negligible success. Direct-to-consumer advertising has increased demand for expensive--and according to some experts, unnecessary or inappropriate -prescription drugs. In this special report H&HN examines the pressures that these factors put on hospitals. PMID- 11912993 TI - A voice for readiness. PMID- 11912992 TI - All pumped up over cardiology. AB - Cardiovascular services are one of the few remaining profit centers for hospitals, and as baby boomers age, the need for such care is skyrocketing. A good cardiology program enhances a hospital's reputation and patient volume. However, the pressures to expand and the cost of swiftly changing technology put hospitals that are trying to keep up in a tight squeeze, which raises the question: is the pulse of change in cardiology too rapid? PMID- 11912994 TI - Progressive wheeze, dry cough: what lies beneath? Hodgkin's disease. PMID- 11912995 TI - Quality improvement in end-of-life care. Small-scale innovations can make a dramatic difference. PMID- 11912996 TI - Early intervention in massive pulmonary embolism. A guide to diagnosis and triage for the critical first hour. AB - The diagnosis of massive pulmonary embolism should be considered expeditiously in all patients with unexplained hypotension, syncope, cardiac arrest, or hypoxemic respiratory failure. The presence of right ventricular overload on physical examination or electrocardiogram is an especially important clue. Depending on local expertise and the patient's stability, V/Q scanning, CT angiography, echocardiography, and right heart catheterization can be useful in establishing a diagnosis of pulmonary embolism. Supportive treatment includes oxygen, vasoactive medicines, and sometimes fluids. Although heparin is important in nearly all patients, 70% to 80% of patients also require an IVC filter, thrombolysis, or embolectomy. PMID- 11912997 TI - Optimal management of septic shock. Rapid recognition and institution of therapy are crucial. AB - Septic shock is a common problem in hospitalized patients. Optimal management depends on rapid recognition, aggressive restoration of circulating volume with fluid boluses, initiation of appropriate antibiotic therapy, implementation of adequate monitoring, and meticulous attention to the details of care. Mean arterial pressure should be increased to between 65 and 75 mm Hg as soon as possible to reduce the likelihood of multiorgan dysfunction. Despite these therapeutic maneuvers, however, mortality rates are likely to remain high until the development of therapies that better target the underlying mechanisms of sepsis. PMID- 11912998 TI - When to suspect obstructive sleep apnea syndrome. Symptoms may be subtle, but treatment is straightforward. AB - OSAS is a serious, prevalent condition that has significant mortality and morbidity when untreated. It is easy to diagnose and treat. Treatment improves quality of life and prevents associated morbidity or death. Response to treatment is high. The goal is to keep the airway open during sleep, leading to improved sleep, better oxygenation, and enhanced day-time alertness. The treatment of choice is CPAP, although bilevel positive airway pressure devices may be used in patients unable to tolerate CPAP. Many surgical treatments have proved effective. Drug therapy may be useful. Weight loss and training to avoid the supine sleeping position are helpful for some patients. PMID- 11912999 TI - Type 2 diabetes therapy. A pathophysiologically based approach. AB - The dramatic increase in the number of classes of oral antidiabetic agents has provided physicians with more tools to help patients manage type 2 diabetes. Of course, glycemic control must remain paramount when choosing an oral agent. However, the mechanism of action of an agent, its side effect profile, and the potential for various nonglycemic benefits may help determine which is the best drug for an individual patient. PMID- 11913000 TI - Using the Internet to support evidence-based practice. PMID- 11913001 TI - Stories connect science to souls. PMID- 11913002 TI - Diabetes education for the Native American population. PMID- 11913003 TI - The group as a basic asset to ethnic minority patients with diabetes. PMID- 11913004 TI - Health-compromising behavior and diabetes mismanagement among adolescents and young adults with diabetes. AB - PURPOSE: This study examined the relationship between health-compromising behavior, age, gender, and diabetes mismanagement. METHODS: A total of 107 adolescents and young adults, 12 to 24 years old, with type 1 diabetes were asked to complete a health-compromising behavior scale and a diabetes mismanagement scale. RESULTS: Based upon participants' responses, the study population was divided into 2 different groups: those involved in health-compromising behavior and those not involved in such behavior. A multiple regression analysis was performed using age, gender, and health-compromising behavior as predictor variables and diabetes mismanagement as the outcome variable. Variables that accounted for significant variance in diabetes mismanagement were being female and being involved in health-compromising behavior. CONCLUSIONS: Adolescents and young adults with diabetes appear to be either involved or not involved in health compromising behavior. Being female and being involved in health-compromising behavior were associated with diabetes mismanagement. PMID- 11913005 TI - Training lay health educators to conduct a church-based weight-loss program for African American women. AB - PURPOSE: Community-based lay health educators have been utilized in a range of settings and with a variety of health issues. However, little has been published about the specifics of training lay health educators to effectively deliver community-based programs. This paper describes the training used to prepare volunteer, church-based lay health educators to conduct a community-based weight loss program, and the evaluation of that training. METHODS: After recruitment through their respective churches, volunteer lay health educators were given structured training in how to conduct the PATHWAYS weight-loss program. Program sessions were observed to monitor program delivery, and participation rates and weight loss were evaluated. RESULTS: The lay health educators were highly consistent in their delivery of the program content. Participant attendance was high and virtually all of the participants completed the program. Participant weight loss averaged 8.3 pounds, which correlated with session attendance. CONCLUSIONS: Given training appropriate to the structure of the program and specific to the targeted health behavior, lay health educators can reliably and effectively administer even rather complex programs. PMID- 11913006 TI - Current computer use by diabetes educators. AB - PURPOSE: The purpose of this study was to ascertain diabetes educators' current use of computer technologies. METHODS: The study population consisted of 279 randomly selected certified diabetes educators, all of whom were members of the American Association of Diabetes Educators. Participants were asked to complete the Attitudes Toward Patient Education Technology instrument. RESULTS: Descriptive findings showed that most educators (89.6%) have access to computers and use them primarily for word processing (43%). Nearly 30% of participants had never accessed the Internet, and no significant difference was noted when this variable was categorized by age. In addition, age and gender were not significant predictors of computer use, although age was significantly related to computer ownership. Patient access and organizational issues were the primary barriers to computer use in patient education. CONCLUSIONS: These findings provide further understanding of diabetes educators' computer use and integration of computer technology in the patient education process. PMID- 11913007 TI - Sequencing diet and exercise programs for African American women with diabetes. AB - PURPOSE: This study was designed to compare the effects of 2 programs that present diet and exercise components in a different sequence. METHODS: At an urban YMCA, African American women with type 2 diabetes, aged 30 to 65, were randomly assigned to either 10 weekly sessions about healthy eating followed by 6 weekly sessions about exercise or to the reverse sequence. Sessions consisted of small group discussions and physical activity or food tasting. Primary outcomes were attendance, percent of calories consumed from fat, fruit and vegetable intake, and minutes of exercise per week. Measures were taken at baseline, and 4 and 12 months after the program. RESULTS: The only group difference found at the 12-month follow-up was in diastolic blood pressure. Time effects for both groups combined included an increase in minutes of activity, an increase in vegetable intake, and a decrease in percent of calories consumed from fat. CONCLUSIONS: This study does not provide definitive evidence of which sequence may be best to bring about behavior change. The effects of sequencing difficult behavioral changes such as diet modification and establishing an exercise habit deserve further study. PMID- 11913008 TI - Sense of belonging and new graduate job satisfaction. AB - This study investigated the relationship between sense of belonging and job satisfaction in the new graduate RN. Ninety-five new graduate RNs answered a survey about demographic information and work setting, as well as their satisfaction and sense of belonging in the work environment. McCloskey-Mueller's Satisfaction Scale and a modified version of the Hagerty-Patusky Sense of Belonging Instrument were used. The conceptual framework was derived from Marlene Kramer's work on Reality Shock (1974). Results showed new graduates were most satisfied with coworkers and least satisfied with professional opportunities for advancement. Sense of belonging and total satisfaction were highest in the home healthcare setting. A Pearson r was used to determine relationships between sense of belonging, total satisfaction, and satisfaction sub-scales. Sense of belonging had significant positive relationships with total satisfaction, interaction opportunities, praise, control, coworkers, and scheduling. Possible future research suggested examining how orientation and work group numbers may affect job satisfaction and sense of belonging. PMID- 11913009 TI - LPN role advancement: from blueprints to ribbon cutting. AB - The role of the Licensed Practical Nurse (LPN) in an acute care setting has been a point of debate for many years. Increasing LPN job responsibility has not been widely reported. In this article, the authors describe the process used to advance the LPN role. Emphasized here is the educators' contribution to the process from inception to implementation. PMID- 11913010 TI - The relationship between death-related experiences, death anxiety, and patient care attitudes among AIDS nursing staff. AB - A questionnaire was completed by 197 nurses working in AIDS care facilities to examine the experiences and attitudes of AIDS nursing staff. Correlations (Pearson r) showed a statistically significant relationship between personal death-related experience and death anxiety and attitudes toward dying patients. CNAs reported higher death anxiety scores and rated the negative aspects of caring for dying patients higher than did licensed practical nurses and registered nurses. The evidence supports the development of appropriate training and support programs in death and dying issues that are tailored to meet the needs of different levels of nursing staff in AIDS care facilities. PMID- 11913011 TI - Increasing critical care skills of noncritical care nurses. AB - Critical care courses for noncritical care nurses are developed to meet staff nurses' needs for additional skills because of increasing patient acuities. To be successful it is imperative that staff development nurses provide input to ensure that course content is directed to meet the needs of nurses in their facilities: hospitals, nursing homes, and home care settings. PMID- 11913012 TI - Compact disc interactive: a multimedia solution for staff education. AB - The findings acquired from two focus groups (13 generalist RNs and 13 specialist RNs) which were held to evaluate compact disc interactive (CD-i) as a self learning option for staff development were reviewed. The findings from these groups were that the CD-i platform offers educators the opportunity to design programs that meet nurses' learning needs without the need for multi-media, computers, and the associated platform training and education. This study reinforced the previously identified problem that nurses have difficulty balancing the responsibilities of providing direct patient care and the need for additional education and training and that self-study is an acceptable alternative to the traditional lecture format. PMID- 11913013 TI - Expanding the role of nurses in Armenia. AB - The dissolution of the Soviet Union and the declaration of Independence by the Republic of Armenia created the need for significant changes in the healthcare delivery system in Armenia. The desire to raise the level of health care presented challenges and opportunities for nurses within the Republic. Members of the departments of nursing at Boston City Hospital/Boston Medical Center in Boston, Massachusetts, University of Massachusetts Medical Center, Worcester, Massachusetts, and the Emergency Scientific Medical Center of Yerevan, Armenia, joined forces through a grant written by Boston University School of Medicine and sponsored by the American International Health Alliance under a cooperative agreement with the United States Agency for International Development to expand the role of nursing. This article describes the assessment, planning, implementation, and evaluation of changes to the role of nursing and the development of new roles for nurses within a hospital in the capital city of Yerevan. PMID- 11913014 TI - Pyramids to success: laying the foundation for professional practice. AB - Communication plays a vital role in an increasingly complex healthcare environment. Quality patient outcomes and successful collaboration depend upon the healthcare provider's ability to communicate needs. A self-directed work team at a large southeastern hospital used a quality improvement process to design a new series of workshops to enhance staff communication skills. The redesigned programming reduced duplication of education content and built upon a conceptual foundation that cultivates professional development. The workshops feature content such as self-awareness, personal effectiveness, and leadership of others. PMID- 11913015 TI - Developing computer-based training for age-related competencies. AB - Competency in adapting care to the developmental needs of patients is not only a clinical expectation but also a requirement of regulatory agencies. Training programs on age-specific care competencies vary widely in format, content, and method. This article describes the process used by the authors to develop computer-based training (CBT) programs on age-specific care competencies and the process used to develop traditional self-paced learning modules on age-specific care competencies. The conversion of these printed modules to CBT is described and reports are given on the pilot test experiences of using CBT with clinical staff. Also included are lessons learned since organization-wide CBT deployment for age-specific care competency and other mandatory training. PMID- 11913016 TI - Utilization-focused evaluation: evaluating the effectiveness of a hospital nursing orientation program. AB - Providing comprehensive orientation programs that prepare nurses for their role as staff nurses is an integral aspect of retention. Therefore, it is vital that staff development educators assess the effectiveness of their nursing orientation programs. In this article, the authors employ a utilization-focused evaluation format to assess a nursing orientation program and, based on the results, offer recommendations for strengthening such programs. PMID- 11913017 TI - Achieving department safety: a team approach to development of a department safety plan template. AB - Hospitals develop safety plans to teach employees to work safely with hazards, to maintain a safe patient care environment, and to enable appropriate response to emergencies affecting the healthcare facility. This article explains the process used to create the Department Specific Safety and Infection Control Plan at United Hospital, St. Paul, MN. PMID- 11913018 TI - Nursing assistants' perceptions of barriers to nutrition care for residents in long-term care facilities. AB - The purpose of this study was to assess nursing assistants' perceptions of barriers to feeding nursing home residents. A 38-item questionnaire was developed from data obtained during four different focus groups. Results suggest that workload and supervision constraints, poor nurse-nursing assistant interactions, inadequate food quality and quantity, and staff educational needs may all negatively influence resident food intake. PMID- 11913019 TI - Preparing for an international consultation. AB - This article outlines the experiences of a staff development educator in preparing for an international staff development consultation. Sections on preparing for travel and for the actual consultation visit provide suggestions in each of these areas. The consultation is described in terms of work done before, during, and after the visit. This information will be helpful to staff development educators considering an international consultation. PMID- 11913020 TI - Using the nursing process to implement a Y2K computer application. AB - Because of the coming year 2000, the need was assessed to upgrade the order entry system at many hospitals. At Somerset Medical Center, a training team divided the transition into phases and used a modified version of the nursing process to implement the new program. The entire process required fewer than 6 months and was relatively problem-free. This successful transition was aided by the nursing process, training team, and innovative educational techniques. PMID- 11913021 TI - Look, learn, and be satisfied: video playback as a learning strategy to improve clinical skills performance. AB - The study investigated the viability of video playback as a means to maintain and enhance nursing skills in today's healthcare environment. The purpose was to determine if video playback would improve performance of psychomotor clinical skills and increase participant satisfaction with the leaning process. A two group sample of registered and student nurses participated in the study. Each participant performed a nursing procedure. Two nursing procedures were used, one for each group. Each nursing procedure was recorded on videotape and played back to the participant who then repeated the procedure and completed a satisfaction assessment questionnaire. Results showed improved performance and learner satisfaction. PMID- 11913022 TI - The value of training and orientation programs in large medical organizations. AB - This study was conducted to measure perceptions of program participants in a large medical center in West Palm Beach, Florida, and to determine the degree to which their participation in the training and orientation program led to 1) greater workplace effectiveness and 2) reduced employee turnover. The results revealed that well-run employee training and orientation programs are not only cost effective, but they also add to the overall quality of life and work in organizations. PMID- 11913023 TI - Mandatory education via the computer: cost-effective, convenient, and creative. AB - With reduced budgets, few resources, minimal time allotted for education, and a long list of topics to cover, staff development and inservice educators often find it difficult to implement cost-effective, comprehensive mandatory education programs. Upper Valley Medical Center's annual mandatory education program via the computer was created to address these issues and resulted in substantial cost savings as well as increased employee satisfaction. PMID- 11913024 TI - Nurses' attitudes and practices regarding voluntary continuing education. AB - This article describes registered nurses' attitudes and practices regarding voluntary continuing education and identifies the factors that influence their decision to participate or not participate. This information is essential for nurse educators in order to meet the education needs of staff. PMID- 11913025 TI - Games galore. AB - Gaming is an interactive process that lends itself to the acquisition and application of cognitive, affective, and psychomotor knowledge and skills. The purpose of this article is to identify the rules, game pieces, and players to assist nursing educators to spin the Wheel of Fortune and enhance their training techniques. PMID- 11913026 TI - Teaching strategies for foreign nurses. AB - To assist foreign-trained nurses to improve their nursing care and enhance outcomes in nursing practice, educators must have specific teaching strategies to build the nurses' self esteem, improve their openness to explore new ideas, and help them obtain new knowledge. The author discusses specific efforts and teaching strategies to facilitate this process. PMID- 11913027 TI - Identifying critical thinking behaviors in clinical judgments. AB - Continuing education and staff development specialists often collaborate with nursing faculty and managers in evaluating the critical thinking behaviors of new staff and nursing students in clinical judgments. The results of this phenomenological study provide insights into specific objective criteria of critical thinking behaviors of domain-specific knowledge, critical reflection, critical thinking competency, intellectual virtues, and action involvement and improvement. PMID- 11913028 TI - Teaching staff to handle a patient's sexually inappropriate behavior. PMID- 11913029 TI - On the relation between mental representation and naming in a child with specific language impairment. AB - The naming and drawing responses of a child with specific language impairment (age 5.5 years) were used to test the hypothesis that deficient storage in the mental lexicon plays a role in the naming problems associated with SLI. In confrontation- and repeated naming, the child demonstrated frequent semantic substitutions and occasional phonologic substitutions. Stochastic modelling of his repeated naming revealed storage deficits to be a source of these errors. Comparative picture naming, picture drawing allowed exploration of this storage deficit and revealed that, for some semantic naming errors, sparse semantic representations were clearly at fault but for others, sparse phonological input and output representations played a role. Phonological naming errors, in contrast, were typically associated with strong semantic representations. Clinical, theoretical, and methodological contributions of this cognitive neuropsychological case study are discussed. PMID- 11913030 TI - Speech timing in children after the Lidcombe Program of early stuttering intervention. AB - It is known that operant treatments can control stuttering in children. However, at present it is unknown why such treatments are effective. Changes in the usual way of speaking are frequently observed after behavioural treatments for adults who stutter, and it is possible that operant treatments for children also invoke such changes. To explore this idea, selected acoustic measures of speech timing were made in eight preschool children before and after receiving the Lidcombe Program, which is an operant treatment for stuttering. No systematic changes were detected after treatment. Considering this finding and a previous report, there is no evidence to suggest that the reductions in stuttering that occur with this treatment are related to systematic changes in speech timing or curtailment of language function. PMID- 11913031 TI - Levels of complexity in phonological disorders: evidence from Cantonese. AB - Longitudinal data from ten phonologically disordered Cantonese-speaking boys were analysed for distinctive features (manner and place). The children labelled 95 pictures, attempting each initial Cantonese segment at least five times. The applicability of Dinnsen et al.'s implicational hierarchy to this data was examined. Categorization of each child's system according to an implicational hierarchy was successful for nine of the ten children when phonetic inventories were considered. In addition, a phonemic inventory based on Dinnsen et al.'s phonetic inventory captured the system of phonological contrasts used by eight of the ten children. The patterns of the children who did not match the hierarchy were considered deviant rather than delayed. The ability of the hierarchy to predict the route of development in this group of children was also examined. The hierarchies (both phonetic and phonemic) successfully predicted the route of change in these cases. Implications for the use of an implicational hierarchy in phonological assessment and treatment are discussed. Further research on feature based phonological development, in both typically-developing and disordered phonological systems is required. PMID- 11913032 TI - Fast mapping of verbs by children with specific language impairment. AB - Two studies examined preschoolers' ability to assign verb interpretations to nonsense words encountered in conjunction with novel actions. Experiment 1 examined the ability of children with specific language impairment (SLI) and younger, normally developing peers to glean a very interpretation when the name of the figure performing the novel action was already known. The two groups of children performed in a similar, accurate fashion. Experiment 2 required preschoolers to rely exclusively on morphosyntactic information to determine whether the novel word represented an object or action. When provided with redundant morphosyntactic cues, children with SLI and language- and age-matched peers succeeded in identifying the novel words that referred to objects but not those that referred to actions. Only the age-matched normal peers were above chance levels when a noun interpretation depended on a single grammatical morpheme (e.g., 'We want the koob' versus 'We want to koob'). The findings suggest that preschoolers, whether or not they have language impairment, have difficulty using morphosyntactic information to bootstrap verbs. Furthermore, redundant but not single morphosyntactic cues facilitate the bootstrapping of nouns. PMID- 11913033 TI - Biotech's next big blowup? PMID- 11913034 TI - Bone vivant. PMID- 11913035 TI - Biotechnology. New research is an easier cell. PMID- 11913036 TI - Coalition for uninsured renews lobbying efforts. With billions of dollars at stake, group launches advertising campaign to draw attention to cause. PMID- 11913037 TI - Shift in priorities. Facing budget cuts, QIOs (Quality Improvement Organizations) to trim hospital services. PMID- 11913038 TI - Working together. Alegent finds a balance with acquisitions. PMID- 11913039 TI - How we can expand coverage. Extending health insurance to millions of uninsured is a matter of collective will. PMID- 11913040 TI - How secure is the safety net? Public hospitals learn to survive in an increasingly tight market by closing, building, replacing and sometimes converting. PMID- 11913041 TI - New views, new vows. ACHE conference realigns agenda: disaster preparedness, staffing top list. PMID- 11913042 TI - Remote control. Specialists are running intensive-care units from remote sites via computers, and at least one health system with the e-ICU is reaping financial rewards--and saving lives. PMID- 11913043 TI - By the numbers. Top 25 hospital systems. PMID- 11913044 TI - 20/20 hindsight. Months after anthrax claimed the lives of several. Americans, hospitals review their reaction to the event--and plan for future crises. AB - Last fall, after a nation suffered the unthinkable attacks of Sept. 11, a second wave of terror held Americans in its grip. Questions still surround a series of baffling cases of anthrax that shook New York and Connecticut. Each of the hospitals involved report that despite the unsolved mysteries posed by these events, they are more cautious and better-prepared for future bioterrorism attacks. PMID- 11913045 TI - [High and low risk human papilloma virus infection in women with CIN. Differential characteristics]. AB - The objective of this study was to determine high risk human papillomavirus infection (HPV-RH) and factors with cervical intraepithelial neoplasia appears (CIN). MATERIAL AND METHOD: From October 1998 to January 2000, a case-control study, was made; women with benefit package from Mexican Institute of Social Security. The cases were of the colposcopic clinic of the department of the Hospital Obstetrics and Gynecology Luis Castelazo Ayala, women histologically diagnosed with colposcopy and CIN cervical biopsy, and controls patients with negative cervical uterine cytologic study of the Preventive Medicine Department, Unit of Familiar Medicine No. 8, of Mexico City. Trained personnel obtained information about socioeconomic and reproductive factors did the interview. A cytobrush was used to take the cervical sample for HPV-RH to determine HPV-RH utilizing Hybrid Capture II test. Both bivariate analysis and logistic regression analysis were used for the adjustment of variables. RESULTS: We analyzed 102 cases and 192 controls, 79% (44/56) of the cases with CIN I and 89% (37/42) of CIN II-III as 21% of controls, respectively, were positive for HPV-RH. Global risk for HPV-RH association to CIN was OR = 40.6 (95% CI, = 17-96.8). Women age was determinative for HPV-RH association to CIN. We observed a high correlation between HPV positive magnitude and CIN degree. CONCLUSIONS: Frequency of RH-HPV in controls and CIN I is higher than other reports in the literature. HPV was identified as the most important agent associated with this neoplasia, other factors involved and age is an important modifier in HPV-RH and CIN. PMID- 11913046 TI - [Angiogenesis in reproductive physiology. Follicular development, formation and maintenance of the corpus luteum]. AB - The process of new capillary formations from previously existing mature vessels in healthy individuals has been mainly studied during cycles of the female reproductive tract. This new capillary formation, known as angiogenesis is related to endogenous regulators that both stimulates or inhibits it. Knowledge on the role of both stimulators and inhibitors under physiological and pathological conditions accentuates a main role of VEGF, FGF, angiogenin and angiopoietics among the formers; and main role of angiostatin and endostatin among the latters. In recent years, angiogenesis in the ovaries that leads to follicular and luteal growth and development has been extensively studied. Whether a number of endogenous stimulators and inhibitors have been identified, the molecular link between the endocrine and vascular system is not fully understood. Therefore, efforts to formulate questions that need answers must be made. PMID- 11913047 TI - [Educational needs in perinatal health among mothers of pregnant adolescents]. AB - This presents the results of a descriptive, transverse study of 95 mothers of pregnant adolescents users of the Instituto Nacional de Perinatologia who were applied an interview to know formally their educational necessities in that situation, since, in spite of its good faith or intention, their advice, actions or the necessary information can be insufficient or inadequate. Programs to guide the parents during the pregnancy of their daughters are practically nonexistent. The purpose of the study is to have information to design an educational strategy that contributes to strengthen the psychosocial net of support for the pregnant adolescents. It was found that most of the parents have labor activity low renumerated, with scholarship average smaller to 10 years; near the 0.40 they are not married, and that the predominant maternal occupation is housewife. It was documented that the average age when the mothers had their first pregnancy it was 20 years. Their general knowledge on reproduction and associate risks showed deficiencies, especially in opportune identification of signs of alarm of the pregnancy and family planning. The preexistence of myths and risky popular beliefs are frequent. Most accepted not to have information to offer appropriate support to their daughters during the pregnancy. A 0.86 accepted that such an event caused diverse changes in the family dynamics. A 0.90 showed good disposition to attend educational courses. PMID- 11913048 TI - [Evaluation of the program for timely detection of cervical cancer in Durango, Mexico]. AB - In Mexico, more of 36% of malignant tumors in the women are cervical cancer, representing an important public health problem. Although cytologic screening for cervical cancer was introduced in 1974, the mortality rate for this disease has been increasing. The state of Durango represents the second national place in mortality by CaCu (3.4/100,000). Since there are few the studies of DOC program evaluation in Mexico were considered important to evaluated the cervical screening program (coverage, assiduity, diagnoses and pursuit) in the state of Durango. Of 40,000 active sexually women attended in the IMSS; we received 11,185 slides during May of 1999 to April of 2000 for cytologic screening. The coverage in this population was 27.96% (11,185/40,000), 8,187 women (73.2%) had cytologic control at least previous two years, 652 (5.83%) had more of four years without control and 2,346 (21%) assisted for first time. The cytological diagnostic showed 189 abnormal Pap (1.68%), and only 40 of them (21.16%) had cytohistopatholgic pursuit and clinic treatment. According to these results we concluded that DOC program in Gomez Palacio, Durango has a deficiency of coverage (72.04%) and pursuit (78.84%). These results indicated the need for development institutional activities of prevention for increase efficiency of preventive services. PMID- 11913049 TI - [Fifty-five years ago. Physiopathology of ectopic pregnancy]. PMID- 11913050 TI - [Staging surgery with minimal axillary invasion in breast cancer. Value of the sentinel lymph node]. AB - Traditionally, axillary dissection has been the best way to staging patients with breast cancer, nevertheless in the early stages (I and II), without palpable axillary nodes (NO) half will not have nodal metastasis and will have been unnecessarily exposed to the morbidity of the procedure. The sentinel node (SN) has showed to be a predictor of the histologic state of the other axillary nodes, its identification allows to adequate staging of NO breast cancer patients avoiding the axillary dissection. OBJECTIVE: To get to know the SN value as predictor of axillary nodal metastasis. MATERIAL AND METHOD: From 2000 to 2001, 62 patients in stages I and II of breast infiltrating ductal carcinoma NO, have been subject to lymphatic mapping and biopsy of the sentinel node with blue dye, the sentinel node was studied intraoperatively and, independently from the result, axillary dissection was practiced to all the patients. A sensitivity and specificity test of the SN as predictor of the histologic state of the other nodes was carried out. RESULTS: In 58/62 (93.5%) the SN was identified, 18/58 patients (31%) had nodal metastasis, only one patient showed false negative SN (92% sensitivity). With the procedure, 98% of the patients were appropriately typified by stages, the specificity of the intraoperatively study is 100%. None of the patients with SN negative had metastasis in other axillary nodes. CONCLUSIONS: The SN predicts, with high sensitivity, the histologic state of the other nodes, appropriately staging the NO breast cancer patients and can prevent more than half of axillary dissections. PMID- 11913051 TI - Enzyme-potentiated desensitization (EPD) is back. PMID- 11913052 TI - Tympanosclerosis (cosmetic) and retraction. PMID- 11913053 TI - Endoscopic view of a nasoalveolar cyst. PMID- 11913054 TI - Sessile vocal fold polyp, contralateral reactive vocal fold mass, anterior glottic microweb. PMID- 11913055 TI - Sinonasal hemangiopericytoma. PMID- 11913056 TI - A patient who had difficulty moving her head and looking at a computer screen. PMID- 11913057 TI - Candida esophagitis. PMID- 11913058 TI - Appeal rejected claims immediately or count on being denied proper reimbursement. PMID- 11913059 TI - The radiologic work-up in thyroid surgery: fine-needle biopsy versus scintigraphy and ultrasound. AB - To compare the effectiveness and predictive value of radiologic studies with fine needle aspiration biopsy (FNAB) in correctly diagnosing thyroid lesions, we reviewed the medical records of 441 patients who had been treated surgically for thyroid disease from 1987 through 1999. We compared the results of thyroid scintigraphy, ultrasound, and FNAB with findings on final surgical pathology. The data were analyzed according to the chi-squared (chi 2) test. Of 189 thyroid scintigraphy scans that showed a hypofunctional (cold) nodule, 52 (27.5%) were found to be cancerous, and of 35 hyperfunctional (hot) nodules, two (5.7%) were malignant (sensitivity = 91%; specificity = 19%; accuracy = 38%; chi 2 = 7.67; p = 0.006). Of 66 ultrasounds that detected a solid or a mixed solid-cystic mass, 16(24.2%) were cancerous, while none of the eight sonograms that showed a purely cystic lesion was malignant (sensitivity = 100%; specificity = 14%; accuracy = 32%; chi 2 = 2.47; p = 0.116 [not statistically significant]). Of the 119 patients whose FNABs were diagnostic, 55 biopsies revealed follicular cells. Of the remaining 64 biopsies, cancer was correctly predicted in 35 of 44 patients (79.5%) and benign disease was correctly diagnosed in 18 of 20 patients (90.0%) (sensitivity = 95%; specificity = 67%; accuracy = 83%; chi 2 = 27.3; p = 0.00). We conclude that in the evaluation of thyroid lesions, FNAB is superior to imaging studies, which yield a relatively high rate of false-positive results. PMID- 11913060 TI - Sentinel lymph node biopsy in SCC of the head and neck: a major advance in staging the N0 neck. PMID- 11913061 TI - Enzyme-potentiated desensitization in otolaryngic allergy. AB - This is a preliminary report of a new method of treating otolaryngic allergy with enzyme-potentiated desensitization (EPD). The nature of EPD and its use in otolaryngology are described. Thirty-six patients have been treated and followed in a private medical practice since February 1997. This article reviews the clinical features of EPD and provides six cases as examples; the clinical features described include allergic rhinitis, serous otitis media, asthma, dermatitis, fixed food allergy, and Meniere's disease. EPD is an effective technique for the treatment of otolaryngic allergy and offers advantages over conventional techniques. PMID- 11913062 TI - Homograft microlathed femur prosthesis in stapedectomy. AB - The use of homografts in ossiculoplasty has been well documented in the literature. In the early 1980s, nonossicular homograft otic capsule bone was used as a prosthetic material in stapedectomy. We began using homograft femur as a prosthetic material in the early 1990s. In this article, we report the results of a retrospective study of the use of homograft femur prostheses. A series of 300 stapedectomies was performed between Aug. 24, 1992, and Jan. 20, 2000. Total footplate removal with preservation of the posterior crus was our procedure of choice. However, in 116 of these cases, the posterior crus could not be used, and a homograft femur prosthesis was substituted. For these prostheses, all homograft femurs were obtained from the American Red Cross. All prostheses were prepared in the bone laboratory and stored in the bone bank until needed. After an adequate period of follow-up, we tabulated our results. We found that in 89 of 113 cases (78.8%) available for follow-up, the air-bone gap was completely closed. In addition, the air-bone gap was closed to within 5 dB in 11 patients (9.7%) and closed to within 10 dB in five patients (4.4%). In all, 105 of the 113 homograft femur prosthetic procedures (92.9%) resulted in a successful outcome. PMID- 11913063 TI - Midline nasal and hard palate destruction in cocaine abusers and cocaine's role in rhinologic practice. AB - Intranasal cocaine abuse can lead to destruction of the palate and perforation of the nasal septum. The pathophysiology of cocaine-induced midline destructive lesions is multifactorial and includes local ischemia secondary to vasoconstriction, chemical irritation from adulterants put in "cut" cocaine, and infection secondary to trauma, impaired mucociliary transport, and decreased humoral and cell-mediated immunity. Cocaine abuse should be suspected in patients with a palatal or septal perforation of unknown etiology. PMID- 11913065 TI - Goldenhar's syndrome. PMID- 11913064 TI - Management pitfalls in the use of embolization for the treatment of severe epistaxis. AB - Angiographic embolization for the treatment of severe recurrent epistaxis was added to the traditional treatment options--nasal packing, cauterization, and surgical vessel ligation--in 1974. Since then, clinical experience has shown that this procedure is safe and effective. When epistaxis cannot be controlled with cautery, nasal packing is the most common next step. As such, it is often performed by emergency physicians and other clinicians who are not otolaryngologists. We report two cases in which intranasal neoplasms were obscured as a result of a significant distortion of the normal anatomy. This distortion was secondary to emergency-room treatment of severe epistaxis by repeated nasal packing followed by angiographic embolization. Pre-embolization angiographic studies and subsequent postembolization endoscopic evaluations did not reveal the presence of the occult neoplasms because of the presence of inflammation and edema after treatment. Clinicians should be aware that nasal packing and embolization can obscure the underlying source of epistaxis, and follow-up radiologic studies and endoscopic evaluations are essential to avoid delays in diagnosis. PMID- 11913066 TI - Mouse and human dendritic cell subtypes. AB - Dendritic cells (DCs) collect and process antigens for presentation to T cells, but there are many variations on this basic theme. DCs differ in the regulatory signals they transmit, directing T cells to different types of immune response or to tolerance. Although many DC subtypes arise from separate developmental pathways, their development and function are modulated by exogenous factors. Therefore, we must study the dynamics of the DC network in response to microbial invasion. Despite the difficulty of comparing the DC systems of humans and mice, recent work has revealed much common ground. PMID- 11913067 TI - Haematopoietic cell-fate decisions, chromatin regulation and ikaros. AB - The regulated production of several terminally differentiated cell types of the blood and immune systems (haematopoiesis) has been the focus of many studies on cell-fate determination. Chromatin and the control of its structure have been implicated in the regulation of cell-fate decisions and in the maintenance of the determined states. Here, I review advances in the field, emphasizing the potential role of chromatin in lineage commitment and differentiation. In this context, I discuss Ikaros, an essential regulator of lymphocyte development and an integral component of a functionally diverse chromatin remodelling network that operates from the early stages of haematopoiesis to the mature lymphocytes. PMID- 11913068 TI - Chemokines: agents for the immunotherapy of cancer? AB - Chemokines, a superfamily of small cytokine-like molecules, regulate leukocyte transport in the body. In recent years, we have witnessed the transition of immunotherapeutic strategies from the laboratory to the bedside. Here, we review the role of chemokines in tumour biology and the development of the host's anti tumour defence. We summarize the current knowledge of chemokine-receptor expression by relevant cellular components of the immune system and the role of their ligands in the organization of the antitumour immune response. Finally, we discuss recent findings which indicate that chemokines have therapeutic potential as adjuvants or treatments in antitumour immunotherapy, as well as remaining questions and perspectives for translating experimental evidence into clinical practice. PMID- 11913069 TI - Roles of heat-shock proteins in innate and adaptive immunity. AB - Heat-shock proteins (HSPs) are the most abundant and ubiquitous soluble intracellular proteins. In single-cell organisms, invertebrates and vertebrates, they perform a multitude of housekeeping functions that are essential for cellular survival. In higher vertebrates, their ability to interact with a wide range of proteins and peptides--a property that is shared by major histocompatibility complex molecules--has made the HSPs uniquely suited to an important role in organismal survival by their participation in innate and adaptive immune responses. The immunological properties of HSPs enable them to be used in new immunotherapies of cancers and infections. PMID- 11913070 TI - Autoimmune thyroid disease: new models of cell death in autoimmunity. AB - Autoimmunity to thyroid antigens leads to two distinct pathogenic processes with opposing clinical outcomes: hypothyroidism in Hashimoto's thyroiditis and hyperthyroidism in Graves' disease. The high frequency of these diseases and easy accessibility of the thyroid gland has allowed the identification of key pathogenic mechanisms in organ-specific autoimmune diseases. In early investigations, antibody- and T-cell-mediated death mechanisms were proposed as being responsible for autoimmune thyrocyte depletion. Later, studies on apoptosis have provided new insights into autoimmune target destruction, indicating the involvement of death receptors and cytokine-regulated apoptotic pathways in the pathogenesis of thyroid autoimmunity. PMID- 11913071 TI - Alcohol, host defence and society. AB - Impaired health caused by alcohol abuse has been known throughout recorded history. Over the past century, alcohol abuse has been clearly linked to host susceptibility to infectious disease, particularly bacterial pneumonia. Recently, both acute and chronic alcohol intake have been shown to result in specific defects in innate and adaptive immunity; these could, in principle, be subjected to specific modulation to overcome the immunosuppressive effects of the most commonly abused substance in the Western world. PMID- 11913072 TI - Ethics of clinical research in the developing world. AB - Many commentators believe that all clinical-trial participants must receive a level of care equivalent to the world's best. Using HIV/AIDS research as an example, we show how this 'Uniform Care Requirement' can undermine biomedical research aimed at improving global health, and then we point towards a more rational and balanced approach to ethical assessment. PMID- 11913073 TI - Mothers and murder. PMID- 11913074 TI - [Herpes: the consensus]. PMID- 11913075 TI - [Risedronate, a new diphosphonate in osteoporosis and Paget's disease]. PMID- 11913076 TI - [Acute coronary syndromes in patients treated with HIV protease inhibitors]. AB - INTRODUCTION: Cardiovascular complications occurring in patients infected by the human immunodeficient virus (HIV) have considerably changed since the appearance, in April 1996, of highly active antiretroviral tri-therapy (HAART), associating reverse transcriptase and protease HIV-1 inhibitors. The spectacular efficacy of anti-proteases has led to the almost complete disappearance of these opportunistic complications. However, in May 1998, acute coronary accidents were reported in the literature, thus questioning the possible responsibility of antiprotease treatment in the occurrence of accelerated atheroma. METHOD: We report a series of 8 seropositive patients in whom an acute coronary event had occurred between February 1997 and February 1999. RESULTS: The patients were young and all exhibited cardiovascular risk factors (smoking, dyslipidemia) and were treated with HIV-1 protease inhibitors. Six patients presented myocardial infarction, one patient unstable angina and one patient effort angina. COMMENTS: A rise in triglycerides was observed principally on ingestion of ritonavir and a rise in cholesterol and LDL-cholesterol with all the antiprotease agents. Glucose intolerance was observed with indinavir. The occurrence of acute coronary events appeared to be related to antiprotease treatment (at the origin of metabolic disorders, endothelial dysfunction...), although it was impossible to say whether the antiprotease agents were responsible for the early atheroma or whether they simply contributed to the event. The coronary lesions were characterized by their number (single artery) and their topography (proximal or median). Nelfinavir may carry less cardiovascular risks than the other antiproteases. Mean term prognosis was relatively good, after therapeutic adjustment (change in antiprotease, strategic measures against cardiovascular risk factors, introduction of anti anginal treatment...). CONCLUSION: Larger and longer studies would help to specify the role of antiproteases in the occurrence of early coronary events. Rigorous monitoring (lipid and glucose measurements, tests to search for myocardial infarction,..) together with the development of new antiretroviral molecules would reduce the number of coronary events in this type of patient. PMID- 11913077 TI - [Outbreaks due to respiratory syncytial virus and influenzavirus A/H3N in institutionalized aged. Role of immunological status to influenza vaccine and possible implication of caregivers in the transmission]. AB - OBJECTIVES: Report of epidemiological, clinical and virological data collected from the prospective surveillance of febrile episodes observed in aged residents of a long-stay care unit of 33 beds, at the University Hospital of Saint-Etienne, during the 1997-1998 winter season. METHODS: Systematic collection of clinical and biological data from febrile patients (> or = 38 degrees C) on a form, including virological findings obtained from a nasal swab and paired serum specimens. RESULTS: From 38 patients (37 of them having been vaccinated against influenza in October 1997), 18 febrile episodes were recorded in 16 subjects, including 3 respiratory syncytial virus infections and a late-occurring outbreak (March 1998) of influenza due to a A/H3N2 strain (15 cases, 14 of them virologically confirmed). No death was noted after the influenza outbreak. In 8 of the 9 tested patients with influenza, "protective" titres of antibodies directed towards the hemagglutinin of the vaccinal strain were present by radial hemolysis test three months before the beginning of the outbreak. During the influenza outbreak, the attack rate of symptomatic infection was 45.5% in elderly and 47.5% in healthcare workers (mainly unvaccinated). The occurrence of the first cases in the latter suggests their possible role in the transmission of the virus to the aged. CONCLUSION: This study underlines the epidemic circulation of multiple respiratory viruses during the same winter season in long-stay care facilities, the occurrence of clinical influenza infections in vaccinated patients exhibiting protective antibody titres and the role of unvaccinated healthcare workers in the propagation of influenza in institutionalised aged. PMID- 11913078 TI - [Legionellosis in HIV-1 infected patients. 4 case reports]. AB - INTRODUCTION: Contrary to patients exhibiting cellular immunodepression, legionellosis is rare in HIV-infected patients. We report 4 cases. OBSERVATIONS: Four men aged 41 to 48 were infected by HIV-1. Three of them were treated with antiretroviral therapy. The clinical and biological manifestations were unspecific. Evolution was favourable with adapted antibiotic therapy (erythromycine for three patients and ciprofloxacine for the fourth). COMMENTS: Legionella pneumophila serotype Lp1 remains the principle etiological agent. Frequent use of contrimoxazole in prophylaxis against pneumocystosis and toxoplasmosis, macrolides, rifamycines and fluoroquinolones in the preventive and curative treatment of mycobacterioses, and the relative conservation of monocyte/macrophage function and immune repair with efficient antiretroviral therapy, may explain the low prevalence of this disease in HIV infections. PMID- 11913079 TI - [Spectacular efficacy of colchicine in a beginning cardiac tamponade]. PMID- 11913080 TI - [Rupture of splenic artery aneurysm during pregnancy]. PMID- 11913081 TI - [Myopathy revealing AL amyloidosis]. PMID- 11913082 TI - [Management of cutaneo-mucosal herpes in immunocompetent patients (ocular manifestations excluded)]. PMID- 11913084 TI - [Update on the outcome of kidney transplantation at the Free University of Brussels]. AB - The present analysis shows that the results of renal transplantation at the "Universite libre de Bruxelles" still have improved in recent years: patient and graft survival at 5 years has risen to 99% and 86% respectively during the 1995 1999 period. This progress is primarily related to the use of less deleterious and more potent immunosuppressants. However, graft failures due to causes other than rejection or patient's death remained unchanged. Among transplantations from living related donors, those from parents yield poorer results than those from children or from one-haploidentical brothers and sisters. Four factors influence cadaveric graft survival: immunosuppression, quality of the graft, donor's age and HLA compatibility, in decreasing order of statistical significance. The immunosuppressive protocols used since 1996 appear particularly promising: thanks to them, 3-year graft survival rate presently reaches 97%. PMID- 11913083 TI - [Arterial hypertension secondary to curable causes in adults]. AB - EXTENSIVE AND COSTLY INVESTIGATIONS: Are not warranted in the vast majority of hypertensive patients. Characteristics identifying the patients at risk for secondary hypertension can be used to define the small percentage of patients with hypertension who require more extensive diagnostic testing and management of their condition. Exposure to certain medicines, foods or drugs may cause reversible rises in blood pressure. Renovascular and adrenal diseases cause curable forms of hypertension. IN MANY CASES, THE PATIENT'S HISTORY: Examination and simple tests can detect such exposures and disorders. Checking for secondary hypertension is therefore an early step required for the management of all patients with hypertension, provided it is based on clinical signs and inexpensive tests. This primary screening cannot exclude the possibility of renovascular or adrenal disease in a small number of asymptomatic patients. The risk of missing a diagnosis is acceptable provided that blood pressure is normalized by non-specific antihypertensive treatment. However, more extensive etiologic investigation is required in patients who subsequently develop resistant hypertension. This secondary screening requires imaging and biochemical tests that are not required for primary screening. CORRECTION OF THE CAUSES: Of secondary forms of hypertension may restore blood pressure to normal. The patient's age affects the reversibility of renovascular and adrenal hypertension after etiologic treatment: the younger the patient, the higher the probability of blood pressure normalization. PMID- 11913085 TI - [Palliative care mobile team at a Parisian university hospital]. AB - Ever since 1987 in France, palliative care is developing in hospitals through PCU and mobile teams. The arbitrary split between care and cure could find its origin in the history of the "cancer war" and in the lack of interest of doctors for patients at the end of their lives. The evolution of mindsets in modern societies goes together with the emergence of the individual, modifying at the same time the perception of pain. This concern for the one who suffers explains the strong requirement to relief pain and to attend the passing away persons. Death is a reality that one constantly tries to evacuate because it is disturbing and shows the limits of medicine. The mobile team within the hospital, including different health professionals, aims at preventing pain at each level of the disease, by a patient effort of awareness among the hospital teams. PMID- 11913086 TI - [Genetics of epilepsy and genetic epilepsy]. PMID- 11913087 TI - [Large intraperitoneal leiomyosarcoma. Report of a case and review of the literature]. AB - The leiomyosarcoma are rare tumors, accounting for only 0.1% to 3% of all gastrointestinal malignancies. 150 new cases are diagnosed annually in the United States. The spectacular character of leiomyosarcoma consists in its measurements, its size reaching 35 x 25 x 17 cm, its weight 5.5 kg, and in the asymptomatic character of the tumor. The final diagnosis, although suspect with preoperative diagnostic procedure, is obtained after histologic examination. Complete surgical excision is the best treatment, the role of radiation therapy and chemotherapy are not yet established. PMID- 11913088 TI - [Crohn's disease]. AB - The etiology of Crohn's disease remains unclear. However, a better knowledge of the immune alterations at the intestinal mucosa level allowed to improve the therapeutic management of these patients. The alternatives to the usual corticotherapy are the use of budesonide (a topical form of steroids) in the mild to moderate diseases or the use of immunomodulators molecules such as the anti TNF antibodies (infliximab) in the severe diseases, resistant to conventional medications. In the chronic active diseases that became steroid-dependent, it is recommended to initiate immunosuppressants such as azathioprine or methotrexate. The use of these molecules requires an optimal adaptation of doses and a regular clinical and biological follow-up to prevent drug-related complications. Surgery should be reserved for treating complications. PMID- 11913089 TI - [Mediastinal mass, pulmonary infiltration, and meningitis]. AB - A 42 year-woman suffering from a non-small cell lung cancer, presenting initially as a mediastinal tumor, is hospitalized for fever, headaches and nausea. An aseptic meningitis is diagnosed. The patient died despite the administration of broad spectrum antibiotics and antituberculous agents. The differential diagnoses are presented. PMID- 11913090 TI - [Acute diarrhea in adults]. AB - The cause of an acute diarrhea is often infectious and a lot of pathogens are implicated: bacteria, parasites and viruses. The clinical approach required several steps and induce different advices such as avoiding stool cultures in the absence of fecal leucocytes, supportive therapy with rehydration, diverse empirical and urgent antibiotherapies. The increasing incidence of antimicrobial resistance of enteropathogens argues against the overconsumption of antibiotics. PMID- 11913091 TI - [Contraception with implants]. PMID- 11913092 TI - [Voluntary pharmacologic termination of pregnancy]. PMID- 11913093 TI - [Marc Herlant, "the hypophysis pope" (1907-1986)]. PMID- 11913094 TI - [New active agents. The composition of commercial pharmaceutical specialties in 2001]. PMID- 11913095 TI - [Generics: similarities, bioequivalence but no conformity]. AB - The using of generic forms (GF) is presented as a potential source of budgetary "saving of money" in the field of pharmaceutical expenses. Not frequently prescribed in Belgium, they win a new interest thanks to the recent making use of the "reference repayment". Sale's authorization of GF is controlled by european rules, but some questions about their identity to original medications remain. Do similarities based only upon qualitative and quantitative composition in active molecules, pharmaceutical forms and biodisponibility give us all requested guaranties? Several cases of discordances can appear; the major elements of non conformity are the nature of excipients, notice's contents and the value of biodisponibility studies. However, in term of economy, in the drug market, development of GF appears to constitute an unavoidable phenomenon. PMID- 11913096 TI - Darwinian paradigm to ecology. PMID- 11913098 TI - Non-light based ecosystems and bioastronomy. AB - The possibility of the existence of life beyond planet Earth has always fascinated humans. However, due to certain circumstances such as the failure of the Viking expeditions to detect any sign of biotic activity on Mars, and the understanding that the presence of life would lead to drastic alterations in the atmosphere of the host planet (alterations that have never been detected on other planets or planetoids of the solar system), the belief that our planet is the only planet to sustain life inside the solar system originated. During the last three decades a series of new complex biological communities have been discovered, in the deep sea, inside caves isolated from the external biosphere, and deep inside the crust of our planet, and found to depend on geothermal energy instead of solar energy for their survival. These discoveries give us new evidence and hope that life might exist not only on other planets, but perhaps even in other planetoids of our solar system. Life may exist in regions other than the surface of a planet, and these areas would be extremely difficult to identify. PMID- 11913097 TI - Luteal phase immunosuppression and meat eating. AB - Immunosuppression during pregnancy makes the mother vulnerable to pathogens. Because meat is the principal source of ingestible pathogens, pregnancy raises the costs of meat eating. Natural selection has crafted a mechanism involving changes in nausea susceptibility and olfactory perception that reduces meat consumption during pregnancy. Evidence is presented showing that the luteal phase is marked by both immunosuppression and changes in nausea susceptibility and olfaction; meat consumption may be reduced during this period, suggesting a mechanism similar to pregnancy sickness. Constraints on compensatory increases in meat consumption outside of the luteal phase explain why women eat less meat than men. Meat is the principal target of acquired aversions. Women possess more aversions than men, suggesting that prophylactic mechanisms sometimes result in longstanding dietary changes. Reproductive immunosuppression explains many aspects of dietary behavior and sheds light on factors that may have contributed to gender-based divisions of labor during hominid evolution. PMID- 11913099 TI - Causing a stir: biomolecular mixing. AB - A proposition is made, and an argument presented for the existence of molecular stirrers inside the living cell. These, as hypothesized, are rotating proteins with long arms mixing the cellular milieu, which is essential for various kinds of interactions. There are no proteins found performing this role, yet. Thus this proposition could be of importance for the progress of scientific finding, by being on the look out for such proteins. Protein stirrers could be rotary motors quite fundamental to the living cell and experimental findings that could suggest the presence of such a stirring machinery are discussed. Possible experimental approaches towards testing this hypothesis are presented. PMID- 11913100 TI - A secondary active transport mechanism modelling. AB - A sustained effort in biochemical, biophysical and physiological research is devoted to the characterization of the transfer of ions and molecules between biological membranes and their aqueous environment. The transported molecules include compounds with substantial structural and physico-chemical differences. It is widely assumed that the biological activity of these compounds arises as a result of binding to active sites in membrane-bound proteins, while the lipid background is considered to play a more passive role. In the simple, symmetric, four-state carrier model for transport of a single solute, transport is down the concentration gradient of that solute. However, a simple modification of this model results in transport up the solute concentration gradient. PMID- 11913102 TI - Is the "small world" effect relevant to evolution? AB - Mutations and selection are the driving forces of biological evolution. We model here the simplest case: an evolving population of asexual organisms. We consider two kinds of mutations: point mutations, corresponding to local displacements in the genotypic space, and all the other genotypic rearrangements, equivalent to long-range jumps. We show that a small-world effect is present in evolution: even a small fraction of quenched long-range jumps makes the results indistinguishable from those obtained by assuming all mutations equiprobable. We apply this result to the evolution of a population on a smooth fitness landscape, showing that the equilibrium distribution is a Boltzmann one, in which the fitness plays the role of an energy, and mutations that of a temperature. PMID- 11913101 TI - Biological evolution as an expression of body-plan potentialities. AB - A growing bulk of recent data from different fields as molecular biology, developmental biology, genetics, paleontology and phylogenetics shows that organisms play a more active role in their evolution than what postulated by the random variation-natural selection paradigm of the neo-Darwinian synthesis. Organisms show during development and morphogenesis autopoietic processes which are related to their body-plan potentialities. These potentialities are expressed through regulatory networks in which a plastic genome participates together with proteins and other substances in an epigenetic space. The epigenetic systems which arise from this interaction may be inherited and then assume a significant role in evolution becoming the source of new acquired characters. The acquisition of new traits through the epigenetic systems is influenced directly by environmental cues. If this process is coherent with the environmental demands it co-operates with natural selection in organism adaptation. An outstanding role in this context may be played by phenotypic plasticity if, as emerges in recent views, it may constitute a general basis for genetic assimilation processes. PMID- 11913103 TI - Mobilis in mobile: a probabilistic and chronotopic model of mobility in urban spaces. AB - In this communication we propose an urban mobility model based on individual random walk driven by a chronotopic action with a deterministic public transportation network. In the absence of chronotopoi the mean field analytic results are found in good agreement with simulations on a computer. When the chronotopoi are switched on, they attract people according to a given law and we obtain a sort of diffusive motion. The model can describe many different kinds of dynamical systems, including biological ones. The work is in progress and the next step will be an empirical test in a concrete case. PMID- 11913104 TI - Lack of "hygiene" as a pretext for closing down small food producers. PMID- 11913105 TI - Self-assembly of the first cells in a hydrophobic environment with hydrogen as the energy source. AB - Two fundamental questions in biology are how and where the first cell(s) self assembled under anoxic conditions on the Earth. The possibility is explored that life first self-assembled in a hydrophobic environment in the subsurface protected from radiation with ubiquitous hydrogen as the likely universal energy source. PMID- 11913106 TI - [Hypertension: once primary, always primary?]. AB - Three patients diagnosed with primary hypertension suddenly developed hard-to treat blood pressure after several years of stable blood pressure. One patient, a man aged 48 years, had developed a renal artery stenosis, which had not been present five years earlier. The other two patients, a man aged 57 years and a woman aged 27 years, were diagnosed with an aldosterone-producing adenoma of the left adrenal gland and a pheochromocytoma, respectively. In patients with previously stable blood pressure, sudden derangement may be due to secondary hypertension on top of the pre-existing primary hypertension. A thorough history and physical examination together with limited laboratory investigations usually leads the way to the correct diagnosis. PMID- 11913107 TI - [The initial treatment of patients with differentiated thyroid carcinoma; consensus and controversies]. AB - Differentiated thyroid carcinoma has a low incidence and a relatively good prognosis. As a consequence, treatment protocols are largely based on retrospective analyses of heterogeneous patient groups, as no randomised controlled trials with residual disease and survival as outcome measures are available. In the Netherlands, guidelines for initial therapy are based on a 1987 international consensus meeting. These guidelines involve near-total thyroidectomy with a few exceptions and routine radioiodide ablative therapy in all cases. Recent publications still support the main measures as advised in this consensus. Although randomised trials have not proven that the consensus guidelines improve the prognosis in thyroid carcinoma, the guidelines provide a base for uniform data collection and thus scientific research. They also provide tools for health-practice quality surveys. PMID- 11913108 TI - [The 'Anemia in the midwife practice' standard issued by the Royal Dutch Organisation of Midwives: a risk of not recognizing iron deficiency and hemoglobinopathy]. AB - The standard entitled 'Anaemia in the midwife practice' issued by the Royal Dutch Organisation of Midwives presumes that the only reason for iron therapy in pregnancy is the prevention of adverse pregnancy outcome due to a low haemoglobin level. Pregnant women are screened for iron deficiency anaemia by means of sequential testing of haemoglobin and mean corpuscular volume (MCV). As a result only 10% of pregnant women will receive iron supplements. This practice will lead to a deterioration in the already low iron status of Dutch premenopausal women. As the haemoglobin reference values are lower than hitherto used, only severely anaemic women will be detected. Due to the low diagnostic accuracy of the MCV test the subsequent selection will be an arbitrary one. The standard sets the cut off values for haemoglobin in black women at an even lower level, which will reduce the number of haemoglobinopathies detected in the immigrant population. The non-carriers in this group will run an increased risk of adverse pregnancy outcome if these cut-off values are used. We are strongly in favour of the measurement of haemoglobin, erythrocyte indices and ferritin in early pregnancy. In this way, iron deficiency, iron deficiency anaemia, anaemia due to other causes and haemoglobinopathies, the latter highly underestimated in the standard, can be detected. PMID- 11913109 TI - [The significance of an elevated cobalamin concentration in the blood]. AB - Elevated levels of serum cobalamin may be a sign of a serious, even life threatening, disease. Diseases such as chronic myeloid leukaemia, promyelocytic leukaemia, polycythaemia vera and hypereosinophilic syndrome are often accompanied by markedly elevated levels of cobalamin in the blood. A rise in the serum cobalamin concentration is one of the diagnostic criteria for polycythaemia vera and hypereosinophilic syndrome. In haematological disorders, the increase in circulating cobalamin levels is predominantly caused by enhanced production of haptocorrin. Several liver diseases such as acute hepatitis, cirrhosis of the liver, hepatocellular carcinoma and metastatic liver disease can also be accompanied by an increase in circulating cobalamin. In liver diseases, the increase in cobalamin is predominantly caused by cobalamin release during hepatic cytolysis and/or through decreased clearance of circulating cobalamin by the affected liver. Liver disorders are not an indication for determining the serum cobalamin concentration. However, a coincidentally observed elevated serum cobalamin concentration is reason for further investigation. PMID- 11913110 TI - [Pleural effusion and empyema as complications of pneumonia]. AB - Parapneumonic effusion is observed radiologically in approximately 40% of the patients with a bacterial pneumonia. In most cases the course of the disease is uncomplicated, and the parapneumonic effusion (PPE) resolves with antibiotic therapy. However, in 5-10% of the patients, PPE becomes more complicated (loculation) and the effusion eventually leads to the formation of an empyema if no drainage has been performed. In view of negative impact on morbidity and mortality, it is important to recognise and evaluate a PPE as soon as possible. Intrapleural pus is the only absolute indication for drainage. In all other cases, the risk of a complicated PPE has to be established in the early phase of the illness, based on radiological, biochemical and microbiological parameters of the effusion. Based on these findings one or more of the following therapeutic strategies can be chosen: tube installation with drainage, fibrinolytical therapy, video-assisted thoracoscopic surgery, thoracotomy with or without decortication, or open drainage. Although every PPE needs to be evaluated on an individual basis, an attempt has been made to formulate a strategy that can be used in clinical practice, based on recent literature and expert opinions. PMID- 11913111 TI - [From gene to disease; from a thrombopoietin receptor gene defect to congenital amegakaryocytic thrombocytopenia]. AB - Congenital amegakaryocytic thrombocytopenia (CAMT) is an uncommon cause of thrombocytopenia in children. CAMT is characterised by an isolated thrombocytopenia and the near absence of megakaryocytes in the bone marrow. The gene involved in the development of CAMT has recently been described. In a number of CAMT patients, mutations in the thrombopoietin (Tpo) receptor gene, c-mpl, were found to be the likely cause of the thrombocytopenia and the complete bone marrow failure that most patients develop. Measurement of Tpo plasma levels and a study of the megakaryocytopoiesis in vitro, may add to the diagnosis. At present the only curative treatment is allogeneic bone marrow transplantation. PMID- 11913112 TI - [Diagnostic image (79). A woman with nagging pain in the lower right abdomen. Mucocele of the appendix]. AB - A 58-year-old woman with abdominal complaints had a cystic mass close to the uterus due to a mucocele of the appendix. PMID- 11913114 TI - [Recurrent, itching and creeping skin lesions in (former) travellers to the tropics: strongyloidiasis]. AB - Two patients, a woman aged 24 and a man aged 86, had suffered from recurrent, itchy, linear, creeping skin lesions, notably on the thighs, buttocks and lower abdomen, for 9 months and more than 50 years, respectively. The woman had been in South America, and the man had worked on the Burma railway as a prisoner of war during World War II. In both patients 'larva currens' was observed. The clinical diagnosis of 'strongyloidiasis' was supported by eosinophilia and raised antibody titres against Strongyloides stercoralis in the blood. No larvae could be detected in either patient. Treatment with albendazole, and ivermectin, respectively, resulted in disappearance of the complaints. S. stercoralis is found in many parts of the world. In the Netherlands the major risk groups in which strongyloidiasis should be considered are people from, and visitors to, South-America (Surinam) and South-East Asia (Indonesia, former prisoners of war). PMID- 11913113 TI - [Surgery and referral for subsequent 131I therapy for patients with differentiated thyroid carcinoma in the south-east of the Netherlands, 1983-1996, compared to the consensus guidelines from 1987]. AB - OBJECTIVE: To evaluate the treatment of patients with differentiated (papillary or follicular) thyroid cancer in general hospitals in the south-east of the Netherlands during the period 1983-1996, in relation to the 1987 national consensus recommendations. DESIGN: Population-based, retrospective, descriptive. METHOD: For the period 1 January 1983-31 December 1996, data on the histology, TNM-stage and treatment (hospital, specialist, type of operation, referral for 131I therapy) of all 236 patients with differentiated thyroid cancer were obtained from the cancer registry of the Comprehensive Cancer Centre South, Eindhoven, the Netherlands. The treatment was compared with the recommendations from the consensus meeting in 1987. RESULTS: Data on 219 patients (137 papillary, 82 follicular thyroid carcinoma) treated in the general hospitals in the region were studied; the 17 remaining patients had been referred from outside the region. Patients were treated at all hospitals in the region; the number of specialists per hospital able to treat thyroid carcinoma (internist and/or surgeon) was limited. In total 79% of the patients underwent a (near-)total thyroidectomy, half of them in two phases, and in 12% of the cases combined with regional lymph node dissection. In the majority of cases, surgical treatment was in accordance with the consensus recommendations: 65-100% of the cases per hospital. The proportion of patients referred for 131I therapy varied from 17% to 90%; referral was more frequent in the case of larger tumours and/or metastases. Of the 24 patients with a small papillary carcinoma without metastases, 79% were not referred for 131I therapy. CONCLUSIONS: The recommendations laid down in the consensus meeting in 1987 were known and appeared to be followed for surgical treatment but for subsequent 131I therapy they appeared to be interpreted differently. A review of the consensus guidelines seems to be worthwhile. PMID- 11913115 TI - [Two patients with isolated vasculitis of the central nervous system]. AB - Over the course of time, a 43-year-old woman and a 39-year-old man developed visual, sensory, motor, cerebellar and cognitive disturbances. Initially these occurred episodically, but later became continuous. Isolated vasculitis of the central nervous system was only diagnosed after brain biopsies were taken, upon which treatment with prednisone and cyclophosphamide was instituted. In both patients the symptoms persisted for 23 and 19 months, respectively, after treatment was initiated, but no new symptoms developed. The woman did, however, develop pancytopenia as a result of the cyclophosphamide treatment. Non infectious vasculitis of the small veins and arterioles of the brain is lethal if left untreated. Leptomeningeal and cortical biopsy is essential in establishing the diagnosis in order to rule out other causative diseases. Combination therapy consisting of prednisone and cyclophosphamide for at least one year is recommended. The efficacy of this treatment is unknown due to the rarity of this disease. PMID- 11913116 TI - [Unwanted 'siesta']. PMID- 11913117 TI - [Rivastigmine for Alzheimer disease; evaluation of preliminary results and of structured assessment of efficacy]. PMID- 11913118 TI - [From gene to disease; presenilins and Alzheimer disease]. PMID- 11913119 TI - [Meningitis after superficial dog bite]. PMID- 11913120 TI - Getting (and staying) connected. PMID- 11913122 TI - Clinical trials: the basics. PMID- 11913121 TI - Turning 40. PMID- 11913123 TI - The biogeochemistry of molybdenum and tungsten. PMID- 11913124 TI - Chemical dinitrogen fixation by molybdenum and tungsten complexes: insights from coordination chemistry. PMID- 11913125 TI - Biosynthesis of the nitrogenase iron-molybdenum-cofactor from Azotobacter vinelandii. PMID- 11913126 TI - Molybdenum enzymes containing the pyranopterin cofactor: an overview. PMID- 11913127 TI - The molybdenum and tungsten cofactors: a crystallographic view. PMID- 11913128 TI - Models for the pyranopterin-containing molybdenum and tungsten cofactors. PMID- 11913129 TI - Transport, homeostasis, regulation, and binding of molybdate and Tungstate to proteins. PMID- 11913131 TI - Molybdenum in nitrate reductase and nitrite oxidoreductase. PMID- 11913130 TI - Biosynthesis and molecular biology of the molybdenum cofactor (Moco). PMID- 11913132 TI - The molybdenum-containing hydroxylases of nicotinate, isonicotinate, and nicotine. PMID- 11913133 TI - The molybdenum-containing xanthine oxidoreductases and picolinate dehydrogenases. PMID- 11913134 TI - Enzymes of the xanthine oxidase family: the role of molybdenum. PMID- 11913135 TI - The molybdenum-containing hydroxylases of quinoline, isoquinoline, and quinaldine. PMID- 11913136 TI - Molybdenum enzymes in reactions involving aldehydes and acids. PMID- 11913137 TI - Molybdenum and tungsten enzymes in C1 metabolism. PMID- 11913138 TI - Molybdenum enzymes and sulfur metabolism. PMID- 11913139 TI - Comparison of selenium-containing molybdoenzymes. PMID- 11913140 TI - Tungsten-dependent aldehyde oxidoreductase: a new family of enzymes containing the pterin cofactor. PMID- 11913141 TI - Tungsten-substituted molybdenum enzymes. PMID- 11913142 TI - Molybdenum metabolism and requirements in humans. PMID- 11913143 TI - Metabolism and toxicity of tungsten in humans and animals. PMID- 11913145 TI - Blowing the whistle: the costs of speaking out. PMID- 11913144 TI - Molybdenum nitrogenases: a crystallographic and mechanistic view. PMID- 11913146 TI - Nursing education: a concern of all nurses. PMID- 11913147 TI - Managing anxiety when patients die. PMID- 11913148 TI - An example of professional responsibility for continuing nurse education. PMID- 11913150 TI - Typhoid outbreak: nurses fear spread. PMID- 11913149 TI - An overview of occupational violence. PMID- 11913151 TI - Queensland nurses take action over pay and staffing issues. PMID- 11913152 TI - Nurse practitioners: forging ahead in Victoria. PMID- 11913153 TI - Caring for the patient with systemic lupus erythematosus. PMID- 11913154 TI - Burial plots. PMID- 11913155 TI - Pets of the future. PMID- 11913156 TI - The do or die decade. PMID- 11913157 TI - Health care has a relapse. PMID- 11913158 TI - John Q.: how real is this horror story? PMID- 11913159 TI - Faith in their father? A Time journalist goes home to witness a priest testing his parish's loyalty. PMID- 11913160 TI - Meet the chipsons. PMID- 11913161 TI - Change of heart. PMID- 11913162 TI - Darwinian struggle. PMID- 11913163 TI - Is your doctor too drowsy? PMID- 11913164 TI - Taming the difficult patient. PMID- 11913165 TI - Opening the door on Medicare peer review. PMID- 11913166 TI - Fraud busters have their eye on you (still). PMID- 11913167 TI - Voice recognition programs speak your language. PMID- 11913168 TI - How to find and keep topnotch clinical staff. PMID- 11913169 TI - A gentle goodnight. PMID- 11913170 TI - Options for boosting your disability coverage. PMID- 11913171 TI - Order supplies online? You bet! PMID- 11913172 TI - Recredentialing? It's a two-minute job. PMID- 11913173 TI - How a "crash course" in medicine changed my life. PMID- 11913174 TI - Watch out for these drug scams. PMID- 11913175 TI - Cut out the insurance middleman? PMID- 11913176 TI - The day I learned how to be a doctor. PMID- 11913177 TI - Diabetes, Part 1. Understanding the pathophysiology of diabetes mellitus. PMID- 11913178 TI - Are latex allergies 'disabilities' under the ADA? PMID- 11913179 TI - Pluggin' away at i.v. deficiencies. PMID- 11913180 TI - When trauma masks a medical emergency. PMID- 11913181 TI - 10 pitfalls in airway management. How to avoid common airway management complications. PMID- 11913182 TI - The next agent of terror? Understanding smallpox & its implications for prehospital crews. AB - The possibility that biological weapons, such as smallpox, will be used is no longer unthinkable. Biological agents have been used since antiquity, and the threat that modern weapons will be used has increased since the events of Sept. 11. Despite international efforts to eradicate smallpox, the potential for its use as a weapon of terror is renewed. Prehospital providers should research the possibility of a vaccination program for their respective agencies and decide on its appropriateness under the guidance of the medical director. In addition, EMS personnel must prepare to handle smallpox and other biological terror weapons. Consult your medical director and local Department of Health for guidelines regarding the appropriate response, treatment and transport of patients exposed to smallpox. PMID- 11913183 TI - Crime scene interaction. Provide patient care while preserving evidence. PMID- 11913184 TI - Exercise restraint. Medical & legal issues you must understand to protect the patient & yourself. PMID- 11913185 TI - The power of one. Leadership through passion, persistence. PMID- 11913186 TI - Pushing the envelope to save more lives. From Pantridge to Smith, Jones and you. PMID- 11913187 TI - Be the catalyst. Transforming the bystander into a citizen responder. PMID- 11913188 TI - D is for defibrillation. The grassroots movement to place AEDs in schools. PMID- 11913190 TI - Starting an AED program. Ten steps to success. PMID- 11913189 TI - Liability no barrier. AED programs can reduce legal risk. PMID- 11913191 TI - How an AED saves a life. PMID- 11913192 TI - Habla Espanol? The importance of learning Spanish for women's health professionals. PMID- 11913194 TI - NIAID and Merck collaborate on HIV vaccine. PMID- 11913193 TI - "Calcium crisis" affects American youth. PMID- 11913195 TI - New CPT and ICD-9 codes for fetal pulse oximetry. PMID- 11913197 TI - Advocating safety. "Whistle-blowing" in the U.S. and Canada. PMID- 11913196 TI - Study to investigate cognitive function in type 2 diabetes. PMID- 11913198 TI - Battling obesity & hypertension. Does the DASH diet really reduce blood pressure? PMID- 11913199 TI - Gauging the benefits, risks, and unknowns of hormone replacement therapy. Exploring HRT. PMID- 11913200 TI - Developing a women's health marketing plan. PMID- 11913201 TI - Maternal positioning in labor with epidural analgesia. Results from a multi-site survey. PMID- 11913202 TI - Pregnancy termination. Understanding and supporting women who undergo medical abortion. PMID- 11913203 TI - The thyroid dance. Nursing approaches to autoimmune low thyroid. PMID- 11913205 TI - Nationwide women's cardiovascular health initiative launched. Nursing leaders converge for landmark AWHONN initiative. PMID- 11913204 TI - Creating a postpartum depression support group. Out of the blue. PMID- 11913206 TI - Tijuana Hope. The journey through health crisis.. PMID- 11913207 TI - Why use cost-based fee schedules? PMID- 11913209 TI - 'It's the denominator, stupid'. PMID- 11913208 TI - HIPAA Administrative Simplification Compliance Act questions answered. PMID- 11913210 TI - Hello, .med.pro. An Internet opportunity for physicians. PMID- 11913211 TI - Physicians slow to warm to PDAs. Questions about time savings, ROI remain. PMID- 11913212 TI - Phone talk. The human side of call management. PMID- 11913213 TI - Helping the uninsured: no need to go it alone. Coalitions ease the burden. PMID- 11913214 TI - Physician recruiters. What they will and won't do for you. PMID- 11913215 TI - Know your risk. Developing effective risk contracts. PMID- 11913216 TI - Backbone, benchmark blueprint: the body of knowledge at work. AB - What does it take to be an effective medical practice executive? What are the core competencies, requirements and information essential for proficiency in medical practice management today? The answers are as close as the Guide to the Body of Knowledge for Medical Practice Management. Published by the American College of Medical Practice Executives (ACMPE) in the closing weeks of 2001, the comprehensive Body of Knowledge is available online (www.mgma.com/acmpe/bokguide.cfm) at no charge. The value, as early adopters share, is proving both structural and strategic. PMID- 11913217 TI - Stretch capability physician extenders can expand access cost-effectively. PMID- 11913218 TI - Is total health management for your organization? Covenant Health System case study. PMID- 11913219 TI - Cultures connect in successful medical marriage. PMID- 11913220 TI - 'The readiness is all.' A strategic approach to managed care. PMID- 11913221 TI - Thou shalt not lie. Health care provider liability in allegations of false or deceptive performance representations. PMID- 11913222 TI - Studies of receptors and modulatory mechanisms in functional responses to cysteinyl-leukotrienes in smooth muscle. AB - Cysteinyl-leukotrienes, i.e. leukotriene (LT) C4, D4 and E4, are inflammatory mediators and potent airway- and vasoconstrictors. Two different cysteinyl leukotriene receptors have been cloned, CysLT1 and CysLT2. This report reviews recent data on CysLT receptor characterisation as well as studies of modulatory mechanisms involved in cysteinyl-leukotriene-induced responses. On the basis of functional studies in isolated smooth muscle preparations, the existence of an additional receptor for cysteinyl-leukotrienes is suggested. In addition, cysteinyl-leukotriene responses in pulmonary vessels were regulated by the release of modulatory factors, of which cyclooxygenase products dominated in the arteries and nitric oxide was the main modulator in porcine pulmonary veins. Moreover, the interconversion between LTC4 and LTD4 and the metabolism into LTE4 may represent a major modulatory mechanism in the guinea-pig trachea by deciding which CysLT receptor is activated by the cysteinyl-leukotrienes. PMID- 11913223 TI - Intimate partner violence, culture-centrism, and nursing. PMID- 11913224 TI - Staying alive: a client with chronic mental illness in an environment of domestic violence. AB - This case study of a young, African American mother with chronic mental illness demonstrates the impact of domestic violence on a vulnerable population. The client was economically disadvantaged, socially isolated, stigmatized, and victimized by repeated abuse from her live-in male partner of 7 years. With this overlay of violence in the home, the client experienced a downward trajectory in health, self-esteem, economic status, personal freedom, social relationships, and legal standing. Using a public health model in the context of an urban nursing center, an advanced practice nurse provided case management and outreach services for the client. PMID- 11913225 TI - Barriers to screening for domestic violence in an emergency department. AB - Screening for persons who are victims of domestic violence is not routinely done in emergency departments (EDs) when clients present for care, nor have all health care professionals been educated in ways to ask appropriate questions to assess victimization. Since questions to identify domestic violence are not on admission sheets and documentation is inconsistent, an interdisciplinary team in the ED of an urban medical center conducted a study using five screening questions. Findings revealed inaccurate statistics for domestic violence and significant barriers for staff participation in screening of adult clients. Clinical implications and follow-up educational programs are discussed. PMID- 11913226 TI - Ten rural women living with fibromyalgia tell it like it is. AB - The purpose of this qualitative study was to gain an understanding of the personal experiences of 10 rural women dealing with fibromyalgia based on the theoretical frameworks of adaptation to chronic illness and social support. The data were gathered from a nursing intervention that provided computer-based peer support and encouragement. The women described themes of pain, fatigue, depression, and sleep disturbances; expressed views on the experience of rural isolation; and shared positive philosophies of dealing with this disease. With this understanding, professionals and significant others can compassionately respond to the needs of sufferers of fibromyalgia. PMID- 11913227 TI - Use of prayer among persons with cancer. AB - This study explored how persons use prayer to cope with cancer. Employing phenomenologic methods, 30 informants were interviewed in depth about why, when, and how they prayed, as well as what they prayed for and the outcomes expected. Findings detail how patients use prayer to ease the physical, emotional, and spiritual distresses of illness. A range of approaches to prayer and topics for prayer was observed, often determined by illness circumstances. The article provides a discussion that begins to suggest how these data can inform clinical practice and future research. PMID- 11913228 TI - Developing a health promotion program for faith-based communities. AB - The article describes the partnership formed between community outreach programs, a school of nursing, and hospitals to implement Healthy People 2010 goals in urban, faith-based communities. To date this program has provided health promotion programs to 125 people from more than 18 congregations in the context of their faith setting. The program has allowed congregants to develop ministry strategies to meet health care needs within the congregation and community. The article provides overall program goals, specific lesson plans, and evaluation strategies. Outcome measures include an increase in health promotion knowledge, participant satisfaction, and improved health in congregations. PMID- 11913229 TI - Children exposed to domestic violence: psychological considerations for health care practitioners. AB - This article reviews the psychological impact of exposure to domestic violence on child development. The purpose is to give insight to the following questions: How does the experience of family violence affect a child's perception of the world and relationships with others? What type of coping style might this same child be likely to develop? What factors help protect a child who has been exposed to violence in the home? In addition, the article discusses assessment considerations for health care practitioners and recommends areas for future research and public policy development. PMID- 11913230 TI - Music listening as a nursing intervention: a symphony of practice. AB - This article presents the use of music listening as an effective, noninvasive intervention designed to assist nurses in creating a healing environment to promote health and well-being. Music has demonstrated effectiveness in reducing pain, decreasing anxiety, and increasing relaxation. In addition, music has been used as a process to distract persons from unpleasant sensations and empower them with the ability to heal from within. As nurses develop practice patterns that are evidence based, the use of music listening could become an integral nursing intervention. To develop a guide for using music listening as a nursing intervention, six principles of practice are identified: intent, authentic presence, wholeness, preference, entrainment, and situating the client. PMID- 11913231 TI - [Treatment of traumatic corneal abrasion with contact lens associated with topical nonsteroid anti-inflammatory agent (NSAID) and antibiotic: a safe, effective and comfortable solution]. AB - PURPOSE: To evaluate the benefit of using therapeutic soft contact lenses in association with NSAID and antibiotic drops for the treatment of traumatic corneal abrasions. The impact on patient pain and on the incapacity to work are analyzed. METHODS: The treatment of 176 consecutive patients presenting with traumatic corneal abrasion was evaluated in a prospective study. Therapeutic contact lenses, NSAID drops (diclofenac), and antibiotic drops (tobramycin) without cycloplegia were used. RESULTS: Psychometric evaluation of pain revealed sufficient comfort with this regimen, allowing 123 patients (80.39%) to immediately go back to their occupations. This return to usual occupations varied with the size of the abrasion: it was obtained in 80 patients (93.02%) from the "foreign body" group (size of abrasion < 10 mm2). In the "other etiologie" group, this ability was obtained in 30 of 41 patients (73.17%) presenting an abrasion smaller than 10 mm2, in 6 of 13 patients (46.15%) presenting an abrasion between 10 and 20 mm2, and in 7 of 9 patients (77.78%) presenting an abrasion between 20 and 30 mm2. All the patients with an abrasion longer than 30 mm2 were unable to continue usual occupations. No serious complications appeared during treatment. CONCLUSION: Treatment of traumatic corneal abrasion with a contact lens associated with topical NSAID drops sufficiently reduced the pain to allow 80.39% of the patients to go return immediately to their normal activities. The combined use of soft contact lenses, NSAID drops, and antibiotic drops without cycloplegia appeared to be an effective treatment for traumatic corneal abrasion. PMID- 11913232 TI - [Value of the Bebe-Vision test in the screening of strabismus and anisometropic amblyopia in infants]. AB - PURPOSE: To evaluate the accuracy of the Bebe-Vision test for detecting strabismic, ametropic and anisometropic amblyopia in childhood. METHODS: We screened 199 infants under 20 months of age. The screening consisted of a full orthoptic examination by a trained orthopist (cover test, fixation test), a forced choice preferential looking technique (Bebe-Vision test) prior to cycloplegia to test visual acuity, cycloplegic refraction by retinoscopy, and examination of the fundi. RESULTS: Fifteen infants were abnormal on orthoptic examination (clinical evidence of esotropia and/or limitation of abduction and/or amblyopia). The Bebe-Vision test demonstrated a significant interocular difference on the same side of the suspected amblyopic eye in 3 cases and on the opposite side in 3 cases, and no difference in 9 cases. The monocular Bebe-Vision test was abnormal in 51 cases and there was an abnormal cycloplegic refraction in 33 cases. Statistical analysis of these tests demonstrated a very low sensitivity (42%) and a good specificity (90%) for the Bebe-Vision test in detecting amblyopia related to refractive error. CONCLUSION: The Bebe-Vision test does not reliably reveal strabismic or anisometropic amblyopia and is not recommended as a screening test. Diagnosis should continue to be based mainly on the classical clinical methods. PMID- 11913233 TI - [Choroidal heat shock overexpression in transpupillary thermotherapy: preliminary results]. AB - PURPOSE: To assess retinochoroidal overexpression of heat shock proteins (HSP-70) induced by a transpupillary laser irradiation below the photocoagulation threshold. METHODS: Four pigmented rabbits were anesthetized and TTT was performed on the right eye using a 810nm diode laser (Iridis, Quantel-Medical, France) adapted on slit lamp (spot size: 1.3 mm, duration: 60 seconds; power 92 150 mW). Series of laser impacts were aimed at the posterior pole of the retina. Left eyes were used as control. Twenty-four hours after laser irradiation, a histological study was done on chorioretinal layers. Tissue samples were fixed in formalin and embedded in paraffin. A monoclonal antibody was used to detect HSP 70 immunoreactivity (mouse IgGl, SPA-810, Stress Gen, Canada), followed by a biotinylated goat antimouse antibody (Dako, Denmark), revealed by the avidin biotin complex (Vectastain kit, Vector, USA) and the AEC chromogen. Retinal structures were further identified by HES coloration. RESULTS: The photocoagulation threshold was obtained for laser power at 150 mW. Under this threshold, HSP-70 immunostaining was the strongest for power 127 mW with a staining of some choroidal cells, including capillary endothelial cells. No HSP 70 immunoreactivity was observed on the retina. For the laser power 107 mW, HSP 70 reactivity was observed only in occasional choroidal cells. For the laser power 92 mW, as for nonirradiated eyes, no HSP-70 immunoreactivity was detected. CONCLUSIONS: Transpupillary 810 nm laser irradiation under the photocoagulation threshold induces choroidal HSP overexpression. This study concludes that choroidal HSP overexpression can be induced during TTT. PMID- 11913234 TI - [On-screen work and visual fatigue and its course after ophthalmologic management]. AB - INTRODUCTION: The number of staff subjected to significant amounts of on-screen work with visual complaints is currently increasing. OBJECTIVE: This epidemiological study aimed to observe the influence of changes in working conditions and an ophthalmological treatment on vision and eye complaints due to on-screen work. MATERIALS AND METHODS: This comparative study included a transversal initial study on the visual symptoms and function as well as a longitudinal follow-up of the efficacy of the recommendations given by the company medical officer. The studied subjects were be exposed to screen irradiation for at least 4 hours a day during the study. The control group was selected according to the age, sex, and education level of the investigated group. RESULTS: We examined 814 subjects under investigation and 325 control subjects. The subjective signs of visual fatigue with on-screen work (burning or stinging eyes, diplopia or blurred vision after work, tearing, globe heaviness and fatigue, and headaches, the latter particularly in female subjects) were significantly more frequent in the exposed group than in the control group. A statistically significant correlation was found with the following factors: poor working conditions, frequent keyboard data entry, far vision optic correction, far vision significantly more often corrected to less than 10/10 at least in one eye, and, finally, near vision exophoria more than 7 diopters. We re-examined 465 exposed and 139 control subjects 1 and 2 years after consultation with the company medical officer. Following the recommendations proved to be effective in 50.5% of the employees. Improving vision by changing the optical correction showed the strongest statistical relation with the decrease in visual fatigue complaints. Organizational and material improvements also led to positive effects on functional discomfort. DISCUSSION: The far vision of those exposed to screen irradiation was significantly worse than in those who were not exposed. The people from the exposed group had vision correction significantly more often. The probable relation between progressive myopia and on-screen work is not excluded, but further investigation is needed. The proposed measures were effective for preventing visual fatigue. However, 49.5% of the study group had persistent visual symptoms, probably because of not following the recommendations, not having an ergonomic correction, or because modifications of blinking, tear film, and the ocular surface were ot taken into account. CONCLUSION: The coordination between the occupational medicine and ophthalmology departments during this study has significantly reduced visual fatigue. Nevertheless, there is still a considerable number of people from the exposed group with persistent symptoms. PMID- 11913235 TI - [Prognosis of retinoblastoma. Report of 50 cases]. AB - PURPOSE: The aim of this study was to analyze the factors influencing the prognosis of retinoblastoma. MATERIALS AND METHODS: In 50 children with retinoblastoma, 69 eyes were reviewed. All patients had a full ophthalmic examination, a B-scan ultrasound, a computerized tomography scan and a pediatric examination. We performed enucleation in 47 eyes (with a histopathological study), external beam irradiation in 16 eyes, curitherapy in 4 eyes, cryotherapy in 3 eyes, and adjuvant chemotherapy in 7 cases. RESULTS: The global survival rate was 87.5%. The main aggravating factors were: the size of the tumor and the extraretinal involvement with extension within the chroid, the sclera, and the optic nerve. CONCLUSION: The prognosis of retinoblastoma mainly depends on the extraretinal invasion. PMID- 11913236 TI - [Value of a new non-contact biometer for intraocular crystalline lens power calculation]. AB - PURPOSE: One of cataract surgery's current imperatives involves refraction: the power of the lens implant must be calculated as accurately as possible. Here we present a new method of biometric ocular measurement using the partial optical consistency interferometer. MATERIAL AND METHODS: This investigation studied the axial length measurement of 100 eyes. Five measurements were taken with a classic echobiometric contact technique using the ultrasonic mode; 5 others were taken with the infrared noncontact technique (IOL Master, Zeiss Humphrey). The latter technique is based on interferometric biometry with optical consistency and measurements were taken with an infrared luminous ray. With extreme rapidity and no contact, the device provides a complete biometry, including axial length, keratometry, and anterior chamber depth. It includes a built-in computer. RESULTS: Comparing the ultrasonic and infrared measurements emphasizes the precision and particularly the high reproducibility of the infrared method. The standard deviations of the samples were significantly lower for the 100 measurements. Its limitations depends on the type of cataract since success was not obtained for certain posterior subcapsular opacities. DISCUSSION: This new method of performing a biometry with a partial consistency interferometer contributes a number of advantages: speed, its noninvasive nature with no contact, the high reproducibility of the exam, as well as precise measurements as shown by the difference in the standard deviations of the two methods. CONCLUSION: Biometry using the optical consistency interferometer seems to be a reliable, reproducible, and precise technique that brings great precision for the calculation of the power of the intraocular implant in cataract surgery. PMID- 11913237 TI - [Porous polyethylene (Medpor) orbital implant. Prospective study of 75 primary implantations]. AB - PURPOSE: To examine the incidence of orbital complications in patients who underwent primary placement of a porous polyethylene implant (Medpor) after enucleation. MATERIAL AND METHOD: Prospective non randomized case series of 75 consecutive patients in whom a porous polyethylene (PP) spherical implant wrapped with homologous sclera was implanted after enucleation. RESULTS: The mean age at the time of enucleation was 42.7 years (range, 1.4 to 80 years). The histopathological diagnoses after enucleation included uveal melanoma in 28 patients, retinoblastoma in 11 patients, phthisis bulbi in 23 patients, neovascular glaucoma in 5 patients, endophthalmitis in 3 patients, ruptured traumatic globe in 2 patients, microphthalmos in two patients, and medulloepithelioma in one patient. Thirty-four patients (45%) had had prior ocular surgery. The prosthesis was fitted after a mean interval of 4.5 weeks (range, 3 to 10 weeks). After a mean follow-up of 20 months (range, 3 to 33 months), there was one case (1%) of conjunctival dehiscence with material exposure secondary to massive postoperative orbital hemorrhage 2 weeks after enucleation. There was no case of orbital cellulitis, implant extrusion, or significant inflammatory response. No PP implant was drilled for peg placement. DISCUSSION-CONCLUSIONS: The anteriorly wrapped porous polyethylene orbital (Medpor) sphere appears to be well tolerated by all age groups with no major complication in primary implantation after enucleation. PMID- 11913238 TI - [Study of visual field and vigabatrin treatment in children]. AB - INTRODUCTION: Concentric visual field defects have been described in association with vigabatrin, a GABA mimetic antiepileptic agent. Few cases have been reported in children. METHODS: A systematic ophthalmological examination was performed in 14 children treated with vigabatrin for seizures. A manual kinetic perimetry test (Goldmann) was done in 11 cases. The ERG was recorded in the 3 cases where perimetry could not be done. RESULTS: All children were asymptomatic. The mean age was 9.6 years. The mean duration of vigabatrin treatment was 41 months. The visual field was abnormal when central and peripheral fields were constricted. A visual field defect was discovered in 6 cases: 4 were severe, 2 were mild. When vigabatrin treatment was stopped, 1 case became worse, 1 case was slightly better, and 1 case remained stationary. A disturbed ERG was found in 3 children (depressed b-wave, raised a/b ratio). CONCLUSION: The visual field defects discovered in children treated with vigabatrin are similar to those described in adults. The incidence and progression of visual field constriction in children with and after vigabatrin treatment are not yet well known. Children treated with vigabatrin should therefore have systematic and regular ophthalmological perimetry, and ERG examinations. PMID- 11913239 TI - [Malignant glaucoma and severe myopia. Report of a case]. AB - Malignant glaucoma is a rare and dramatic complication that occurs more frequently after glaucoma filtration surgery for angle closure glaucoma and rarely for open angle glaucoma. We report a case of a highly myopic monocular 38 year-old woman, with a primary open angle glaucoma who developed malignant glaucoma in the first postoperative days of a trabeculectomy. Treatment consisted of extracapsular cataract extraction and vitrectomy, with the removal of the anterior hyaloid and preventive circular buckling. Late postoperative events have been marked by the occurrence of anterior synechiae and residual hypertension. We discuss the pathogenesis of malignant glaucoma, its management and the results of the different therapeutic procedures. PMID- 11913240 TI - [Gyrate atrophy of late disclosure]. AB - Gyrate atrophy of the choroid and retina is a chorioretinal degeneration caused by a deficiency of ornithine aminotransferase. We report the case of a 29-year old woman who showed an advanced stage of gyrate atrophy of the choroid and retina fortuitously discovered. Examination of the patient provided an opportunity to describe pathophysiology, knowledge on the ornithine aminotransferase gene, and finally the therapeutics used in this disease. PMID- 11913241 TI - [When should the anti-glaucoma treatment start?]. AB - The decision to start a glaucoma treatment in a newly diagnosed patient is easy when the disease is clearly identified with typical optic nerve cupping, visual field defect, and elevated intraocular pressure. However it often appears more complicated, especially in young patients for whom a careful analysis of the fundus and an examination of nerve fiber layers should be assessed to detect an early sign of glaucoma before any visual field change. If glaucoma is present, patients with intraocular pressure above 14 mmHg should be treated, as in such cases there is always a "pressure risk factor". In case of isolated elevated ocular pressure, in France it is usually considered that a pressure above 25 mmHg should be reduced, with a target pressure at least of--20% from baseline. Other factors should also be considered such as quality of life and cost of the treatment. The psychological profile of the patient should also be taken in to account and clear information concerning the risks and potential severity of the disease should be given before the onset of treatment. PMID- 11913243 TI - [Is trabeculoplasty still indicated?]. AB - Trabeculoplasty was introduced in 1979 to decrease intra ocular pressure. Long term results have been unsatisfactory compared to the first publications, because of the repeated relapse of initially successfully treated eyes. However, the technique may be particularly useful in numerous clinical conditions: pseudo exfoliation syndrome, pigmentary dispersion, slight cataract waiting for combined surgery, and in elderly patients. Developing new energy sources (i.e., selective laser trabeculoplasty) could extend the indications of this method, with longer lasting results and/or completely harmless retreatments. PMID- 11913242 TI - [New medical treatments of glaucoma. New strategies?]. AB - Several new classes of ocular hypotonia medications have recently become available. These include the topical treatments carbonic anhydrase inhibitors, the alpha-2 agonists, the prostaglandin analogues, and a fixed combination of beta-blockers and carbonic anhydrase inhibitors. With an equal or superior efficacy than beta-blockers, these new treatments must be reconsidered as first line therapy in glaucoma. The long-term safety profile of antiglaucomatous drugs plays an important role in the new trends of medical treatment. These data, better known and detailed over time by clinical experience, are changing our habits and advancing therapeutic strategies for a more effective treatment, better suited to the individual, thus allowing the best quality of life possible. PMID- 11913244 TI - [When should glaucoma be surgically treated?]. AB - The medical treatment of primary open angle glaucoma has progressively become more and more efficient and safe, and surgery is therefore mostly restricted to failure of and intolerance to antiglaucoma eyedrops. Glaucoma surgery may thus cause severe complications and a high risk of failure has tempered its prognostic. Nevertheless, when efficacious, glaucoma surgery definitively resolves two major pitfalls of medical treatment: patient compliance and eyedrops tolerance. Moreover, new surgical developments of surgery have come from the new technique of non-penetrating deep sclerectomy, which is actually an external trabeculectomy involving removal of the area of maximal resistance to aqueous outflow. This procedure has a very low risk of complications, much lower than that of standard trabeculectomy, but its efficacy is still controversial. Whatever the technique chosen for filtering surgery, antimetabolites may be used in order to limit the risk of postoperative fibrosis, but they also expose to specific, sometimes sight-threatening, complications. Therefore, the most important--and also the most difficult--choice for treating glaucoma patients still remains the best timing for surgery, either excessive, useless, and aggressive medical treatment, or systematic primary surgery. PMID- 11913245 TI - [How to prevent and treat glaucoma surgical failures?]. AB - Although many improvements have been made to filtering surgery in glaucoma, some post operative complications do occur quite often. Patients need careful and frequent follow-up to assess and treat the early complications that may occur after glaucoma surgery. Over the medium and long term, insufficient verification of the intra ocular pressure signals the failure of the initial surgery. In this article, we review the diagnostic tools useful for detecting the early and late failures of glaucoma surgery and we provide recommendations for their treatment. PMID- 11913246 TI - [Large basal cell carcinoma of the external palpebral angle]. AB - A clinicopathologic case of an 89-year-old female patient with a single cutaneous tumor at the outer part of the outer left eyelids is reported. It was a pink nodular mass, well circumscribed, exophytic and with thin vessels. After surgical removal, histopathology showed that the tumor was a typical basal cell carcinoma, with many cystic-like cavities. Basal cell carcinomas can be commonly observed on the eyelids and surgical excision of these small size lesions is an easy way to perform a curative treatment. As often in other tumors, early diagnosis is probably the easiest way to improve the prognosis. PMID- 11913248 TI - Nutrition: impact on oral and systemic health. AB - Good dietary practices and optimal nutritional status promote growth and tissue development, as well as feature prominently in the prevention of diseases. Malnutrition (particularly protein-energy malnutrition, which invariably involves concurrent deficiencies of the antioxidant micronutrients) promotes salivary gland hypofunction, impaired immunity, and an early shift in the oral microbial ecology toward a preponderance of anaerobic organisms. The immune suppression, which includes impaired cytokine function as well as diminished acute-phase response to infections, impacts negatively on the natural history of inflammatory periodontal diseases. The pathogenesis of oral cancer is influenced by deficiencies of antioxidant nutrients, and there is evidence for diminished DNA methylation, disruption of DNA integrity, and increased DNA damage in folate deficiency. PMID- 11913249 TI - Impact factors on development of cirrhosis and subsequent hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC), on the rise in many countries, is of multifactorial etiology. Its etiological associations differ between populations at high and low risk. Africans and Chinese have the highest incidence of HCC, but other affected groups include African Americans, Japanese, and Native Americans. Chronic infections by hepatitis B and hepatitis C viruses are major risk factors worldwide, although mechanisms through which the infections cause liver cancer are yet to be explained. Other documented risk factors have been postulated and include dietary exposure, cigarette smoking, alcohol consumption, diabetes, oral infection, and oral contraceptive use. In addition, many naturally occurring and synthetic chemicals to which humans are exposed via accidental contamination of food or water are shown to induce liver cancer in experimental animals. Consequently, assessment of possible human liver cancer risk associated with such exposures is complex. Early diagnosis and transplantation are the best treatments presently, although transplantation is not widely available due to donor shortage. Every effort should be directed toward the prevention of HCC, through the treatment and prevention of hepatitis and oral infections, prevention of chronic hepatitis progressing to cirrhosis, and prevention of the cirrhotic liver from developing HCC through chemopreventive modalities. However, at present, very few such studies exist. PMID- 11913251 TI - Enhancing oral and systemic health. AB - Much published research documents continuing racial and ethnic disparities in health, particularly for African Americans, which apply to both oral and systemic diseases. Current research suggests biologically plausible associations between oral and systemic diseases; however, clear cause-and-effect relationships have not been substantiated. Some researchers and health care providers have noted anecdotal associations between oral and systemic health, as well as compounding adverse effects of oral and systemic diseases and dysfunctions. Historically, African American physicians, dentists, and pharmacists have bonded together under one organizational umbrella to combat discrimination, prejudice, and racism directed at them and their patient populations. This coming together has resulted in a more comprehensive clinical, behavioral, economic, and public health decision-making process related to the general health and well-being of their patient populations, such as maximizing health care visits, treatment plans, reimbursements, and oral and systemic health care follow-ups. According to the 1985 Secretary's Task Force Report, the six causes of excess deaths among African Americans were: cardiovascular disease and stroke; cancer; diabetes; cirrhosis; homicide and accidents; and infant mortality. In 1991, HIV/AIDS became the seventh cause of excess deaths. This article summarizes salient information about cardiovascular diseases, diabetes, cancer, and the social and behavioral factors related to oral and systemic health. PMID- 11913250 TI - Periodontal disease, race, and vascular disease. AB - Recently, a number of studies have rekindled the possible hypothesis that oral health has repercussions beyond the oral cavity and is associated with systemic diseases. Interestingly, it is a return to an old theory that chronic infections and inflammation played a crucial role in atherosclerosis. This larger theory was advocated by French physicians, among others, at the beginning of the 20th century. In this article, we will review the epidemiologic evidence pointing to a possible association between oral health and vascular diseases and examine the role of race/ethnicity in the interpretation of this association. PMID- 11913252 TI - Exploring interrelationships between diabetes and periodontal disease in African Americans. AB - Population-based data on interrelationships between diabetes and periodontal diseases among African Americans are limited. This article is an overview of our knowledge regarding the bidirectional relationship between diabetes and periodontal diseases and a descriptive analysis of data from the Third National Health and Nutrition Examination Survey (NHANES III), focusing on the diabetes periodontal diseases interrelationship in African Americans. Results of the analysis are consistent with the current body of evidence supporting a bidirectional relationship between diabetes and periodontal diseases and indicate generally poorer periodontal health and glycemic control among African Americans. The results also indicate significantly lower dental care use in dentate African Americans with diabetes than in the US non-Hispanic white population with diabetes. PMID- 11913253 TI - Chronic alcoholism: a common risk factor in oral cancer and alcoholic cirrhosis. AB - Oral cancer and alcoholic cirrhosis are relatively common diseases encountered in medical and dental practices. This article reviews the clinical, pathophysiological, and epidemiological characteristics of these two conditions. A major risk factor common to both oral cancer and alcoholic cirrhosis is the excessive use of alcohol. A challenge for practitioners and researchers is to become mindful of the connection between oral cancer and alcoholic cirrhosis. Earlier studies exploring these relationships and potential mechanisms are discussed. PMID- 11913254 TI - HIV/AIDS: impact on the African American community. AB - The human immunodeficiency virus (HIV) infection has had a devastating impact on the lives of African Americans. In the United States, it is estimated that almost half of all cases of HIV infection in men occur among black men and almost 70% of the cases in women occur among black women. Acquired immunodeficiency syndrome (AIDS), which results from HIV infection, has become a leading cause of death of African Americans between the ages of 25 and 44. Denial regarding how the disease is spread, particularly among heterosexuals, and stigmatization about the disease continue to be barriers to effective prevention campaigns within African American communities. Aggressive cultural and ethnic-specific educational campaigns, focusing on prevention, are needed to curb the current spread of HIV within this population. PMID- 11913255 TI - Customer service revisited. AB - Customer service requires a system, which includes, but is not limited to having pleasant staff members. It requires a systemized approach to every patient every day. The steps of customer service are as integral to the success of the practice as are those of scheduling and case presentation. Customer service is a unique opportunity to differentiate a practice, justify the value for fees, and satisfy almost every patient. PMID- 11913256 TI - Interim implants for immediate loading of temporary restorations. PMID- 11913257 TI - Predictable single tooth peri-implant esthetics: five diagnostic keys. AB - The creation of an esthetic implant restoration with gingival architecture that harmonizes with the adjacent dentition is a formidable challenge. The predictability of the peri-implant esthetic outcome may ultimately be determined by the patient's own presenting anatomy rather than the clinician's ability to manage state-of-the-art procedures. To more accurately predict the peri-implant esthetic outcome before removing a failing tooth, five diagnostic keys are discussed. These keys include relative tooth position, form of the periodontium, biotype of the periodontium, tooth shape, and position of the osseous crest. PMID- 11913258 TI - Immediate total tooth replacement. AB - Successful implant placement at the time of extraction has been documented. Implant placement at the time of extraction was initially performed as a two stage procedure often with barrier membranes and sophisticated second-stage surgical uncoverings. The authors describe the next generation of this technique, including atraumatic tooth removal with simultaneous root form, implant placement, and temporization at one appointment. This technique of "Immediate Total Tooth Replacement" allows for the maintenance of the bony housing and soft tissue form that existed before extraction, while at the same time establishing a root form anchor in the bone for an esthetic restoration. PMID- 11913259 TI - Site development for anterior single implant esthetics: the dentulous site. AB - An inevitable loss of soft and hard tissue after tooth extraction often results in a compromised site for anterior implant esthetics. Various augmentation techniques have been advocated as corrective procedures, but these techniques are time-consuming and not always predictable. Therefore, the fundamental concept of site development for anterior single implant esthetics in the event of a failing tooth is to preserve the existing gingival and osseous tissue. This article identifies factors that affect tissue changes and describes an integration of surgical and prosthodontic approaches that can optimize the tissue maintenance in the anterior esthetic zone. PMID- 11913260 TI - Implant management for comprehensive occlusal reconstruction. AB - Implant prosthodontics is a predictable treatment modality for patients who require occlusal reconstruction. Specific indications include lack of distal abutment, long span segment, compromised natural abutments, and occlusal dysfunction. With proper treatment planning and an understanding of the occlusion and biomechanics of implant prosthodontics, the restorative team can predictably fabricate a functional, esthetic, and comfortable prosthesis. PMID- 11913261 TI - Esthetic soft-tissue augmentation adjacent to dental implants. AB - Patients and clinicians are becoming increasingly aware of the need to improve dental esthetics. After tooth extraction or trauma, there may be residual hard- or soft-tissue defects. These may be related to deficits in the bone or soft tissue. The evaluation of preoperative study casts, radiographs, and photographic documentation can aid the clinician in determining which types of augmentation are necessary. If adequate bone volume is present, the implants are placed according to protocol. Soft-tissue defects can be significantly reduced by implanting various biomaterials between the inner borders of the flap and surrounding bone. This case series describes patients who were successfully treated for soft-tissue defects with either demineralized freeze-dried bone or bovine bone. PMID- 11913262 TI - New tuberculosis vaccines: prospects for reversing worldwide trends. PMID- 11913263 TI - A new composite resin cement: report of a case. PMID- 11913264 TI - The number one issue facing dentistry today. AB - The hiring environment in dentistry will probably not improve significantly over the next few years. You should certainly encourage any dental organizations to support campaigns to attract individuals into dental staff positions through job awareness programs and new training schools or environments. Dentistry is an outstanding profession. Unfortunately, when your office is short staffed, you can not fully enjoy your practice. PMID- 11913265 TI - The finishing touches for more predictable esthetic dentistry. AB - The popularity of esthetic dentistry has transformed the traditional dental practice. Anterior tooth restorations impact the beauty of a smile as well as the function of the entire dentition. The long-term success of esthetic restorations can be improved by controlling the occlusal forces to which they are subjected. During sleep and periods of intense concentration, dental patients move their mandibles far beyond the accustomed positions and are able to place significant forces on anterior teeth. The dentist must test and balance new restorations to ensure that they allow for the full range of mandibular movement. Three basic types of movements are discussed. PMID- 11913266 TI - Finishing and polishing techniques for composite resins. PMID- 11913267 TI - Immediate implant placement on removal of the natural tooth: retrospective analysis of 1,081 implants. AB - Conventional implant therapy dictates a period of 1 to 2 years from the start of treatment to the completion of the restoration. In contrast, immediate implant placement has resulted in the initiation of prosthetic treatment in as little as 3 to 6 months with the additional benefit of reducing alveolar bone resorption, patient morbidity, and expense. This article reports on the criteria for immediate implant placement into an extraction socket, as well as a reliable technique and a retrospective analysis of 1,081 implants placed immediately into extraction sockets. Of the 1,081 implants placed, 35% were followed for 1 year, 46% were followed for 2 to 5 years, and 19% for 5 to 11 years postloading. The criteria for success were complete stability and no further bone loss than had appeared at the second stage surgery and on the day of restoration. The overall survival rate in this current study is 95%. PMID- 11913268 TI - Snoring and sleep apnea: dental and medical perspectives. Interview by Mark J. Friedman. PMID- 11913269 TI - Effects of a nutritional supplement on periodontal status. AB - Among the recommendations for the maintenance of gingival and periodontal health, few have focused on the value of nutritional supplements. The purpose of this study was to compare the effect of certain nutritional and plant-derived nutraceuticals and a placebo tablet in the reduction of gingivitis, bleeding, probing depths, and attachment levels in a 60-day two-cell, randomized, parallel clinical trial for patients with Type II periodontal disease. The vitamin therapy was introduced as an adjunct to patient homecare to determine if there was a quantifiable improvement to soft-tissue health and periodontal damage. Sixty three patients were randomly divided into two groups of 32 and 31 subjects and given either a vitamin tablet containing seven active ingredients (experimental treatment) or a placebo tablet. The clinical parameters assessed were the gingival index (GI), bleeding index (BI), periodontal pocket depth (PD), and attachment levels (AL), and were recorded at baseline and 60 days. Patients took the assigned tablet at breakfast and at dinner after brushing their teeth twice daily. After 60 days, the data showed a clinical reduction in the GI, BI, and PD for the experimental group (P < .0001). There were no significant changes for AL with either the experimental or the placebo group. When the data were further analyzed for pocket depths of > or = 4 mm in patients receiving the experimental treatment, there were clinically significant improvements in the GI and PD from baseline to 60 days (P < .0001), but no significant differences in the BI and AL. There were no statistical differences in any of the indices when the data were compared between men and women. The results of the present study suggest that a multi-vitamin nutritional supplement might be a beneficial adjunct to the required established periodontal treatment. PMID- 11913270 TI - Rapid-setting encapsulated glass-ionomer restorative cement. PMID- 11913271 TI - An innovative tissue-retraction material. AB - One of the most challenging problems of fixed prosthodontics is tissue control. Gingival retraction before a final impression can be very frustrating and time consuming. Many different techniques have been developed over the years to accommodate the clinician's struggle to obtain tissue control and achieve an ideal impression. This article discusses several of those techniques and how the new, innovative product Expa-syl can be incorporated into these techniques. Expa syl is an injectable retraction and hemostatic agent that can cause little trauma to the tissue as well as save the dentist time and money. The author elaborates on the multiple uses of Expa-syl and the correct techniques for making this material a successful tool in any dental office. PMID- 11913272 TI - Expanded clinical uses of a novel tissue-retraction material. AB - Impression making for all fixed prostheses requires access to the prosthetic margin while minimally traumatizing the tissue, so that clinicians can provide as much clinical information as possible to the laboratory technician. This information allows the technician to design the prosthesis to meet the criteria of the periodontium and allow the gingival tissues to recover to their original state. Until now, there have been three traditional options in the armamentarium for retraction procedures: packing cord, electrosurgery, and laser surgery. These procedures all have degrees of trauma that vary based on the experience of the clinician. The gingival tissue may be very delicate and susceptible to recession if too much trauma occurs. This article describes an expanded use of Expa-syl, a newly introduced tissue retraction material, as it relates to minimally invasive tissue management, while still providing an ideal gingival tissue environment for long-term health. PMID- 11913274 TI - Building the boutique practice. AB - To fully educate patients today, dental practitioners must communicate why a practice is unique. Sell your patients on the idea that your practice is the best place for their treatment. Start by informing the current patient base and all new patients about the breadth of your procedures and services. This knowledge will motivate them to become the referral base you need to build your boutique practice. While the boutique practice enables you to provide the highest quality care, highest levels of customer service, and highest fees of any practice in dentistry, perhaps its greatest asset is its ability to help you chart your course through the changing world of dentistry. PMID- 11913273 TI - A predictable gingival retraction system. AB - Predictable tissue retraction for impressions and insertion of restorations can be a laborious task for many dental professionals. The Expa-syl paste retraction system has significantly improved the ability to achieve tissue management even under the most difficult situations and in less time. This article demonstrates the use of this paste retraction system to control tissue for impression making as well as for the insertion of adhesively placed nonmetal restorations. PMID- 11913275 TI - A clinical study evaluating a new chairside and take-home whitening system. AB - This study evaluated the efficacy of tooth color modifications with a 15% hydrogen peroxide in-office whitening system when applied for a 30-, 45-, or 60 minute period and combined with a 10% carbamide peroxide take-home system. Twenty four patients participated in this randomized, parallel clinical evaluation. Six maxillary anterior teeth with a shade of A3 or darker were selected. Patients were randomly divided into 3 groups of 8: Group I patients received a 30-minute application, Group II received a 45-minute application, and Group III received a 60-minute application of the initial in-office treatment. All patients returned at 24 hours for shade evaluation and receipt of a whitening tray with a 10% carbamide peroxide for 7 consecutive days. Patients returned at 24 hours, 72 hours, and on the 8th day of take-home treatment for shade evaluation. Kruskal Wallis ANOVA showed no statistically significant difference between the groups (P > .10) at completion of treatment. When all three groups were combined, a mean shade change > 8 was seen. No significant difference existed for tooth color modifications when varied application times of 30-, 45- or 60-minute in-office whitening with 15% hydrogen peroxide was used in combination with 10% carbamide peroxide take-home whitening. PMID- 11913276 TI - The acrylic occlusal plane guide: a tool for esthetic occlusal reconstruction. AB - A technique has been described that clinically evaluates esthetic problems and transfers that information to the laboratory. This facilitates the fabrication of a diagnostic wax-up, a provisional trial restoration, and a final prosthesis that is functionally and esthetically pleasing. PMID- 11913277 TI - Therapeutic sequence in the management of advanced cases in implant dentistry. AB - Osseointegrated implants have contributed significantly to the treatment of mutilated dentition. Many patients treated with removable prostheses have benefited from implant-supported fixed prostheses. Achieving successful osseointegration is now a predictable procedure and the successful integration of implants is no longer the main objective. Successfully restoring the form, function, and esthetics of the system, as well as maintaining osseointegration, is now a main objective. This goal depends on proper diagnosis and treatment planning, proper case selection, and an effectively and efficiently planned treatment approach. PMID- 11913278 TI - Long-term performance of Osseotite implants: a 6-year clinical follow-up. AB - In this prospective, multicenter study, 147 Osseotite implants were placed in 75 patients (32 men and 43 women with a mean age of 54 years) using a conventional two-stage surgical protocol with 3 months of healing time in the mandible and 6 months in the maxilla. Of the 147 implants, 69% were inserted in posterior sites and 64% were short implants of 10 mm or less. From the time of implant insertion to second-stage surgery, 5.1 months (+/- 2.4 months) elapsed. Restorative treatments included 25 single-tooth replacements (28.4%), 58 short span fixed bridges (65.9%), 1 full-arch reconstruction (1.1%), and 4 overdentures (4.5%). The mean time from implant placement to final recall was 74.1 months (+/- 8.9 months). At second-stage surgery and at 6-month and annual follow-up examinations, implants were evaluated for mobility, peri-implant radiolucency, gingival health, signs and symptoms of infection, neuropathies, paresthesia, and crestal bone levels. A 3-year interim report identified 5 implant failures, 4 of which occurred as a clustering phenomenon in a single, medically compromised patient. Using the life table analysis method, the cumulative implant success rate was calculated at 96.6%. The 3-year interim report indicates that the implants developed an extended, functional osseous state that remained stable for more than 6 years. PMID- 11913279 TI - Anatomical post design meets quartz fiber technology: rationale and case report. AB - Endodontically treated teeth frequently require a post and core to serve as a foundation for the coronal restoration. Remaining tooth structure, physical properties of the post material, post shape, and cement type all contribute to the success of the restoration. Post adaptation to the canal walls also represents an important element in the biomechanical performance of the prosthetic restoration. A double taper post system made of quartz fiber and epoxy was developed to conform more precisely to the shape of endodontically treated canals. Immediate benefits of this post system include minimal tooth structure removal during canal reshaping, greater post-to-canal adaptation in the apical and coronal half of the canal, and good post retention. The use of a quartz fiber/epoxy material with a lower modulus of elasticity also reduces the incidence of root fracture. Furthermore, the esthetic nature of the colors offered with this post system (translucent and off-white) provide a favorable foundation for eliminating discoloration caused by a metallic post placed under all-ceramic crown systems. PMID- 11913280 TI - Versatility of resin composite: esthetic considerations. PMID- 11913281 TI - Long-term clinical performance of CEREC restorations and the variables affecting treatment success. AB - Since the first patient was treated with a CEREC restoration in 1985, several system developments have expanded the indications for which this system can be applied. Concurrently, ongoing research in private practices has contributed to documentation supporting the placement of CEREC restorations based on their performance relative to specific criteria, including tooth vitality, papillary bleeding, quality of margins, quality of ceramic used, treatment required, and failures. This article summarizes the methodology and findings of the ongoing clinical evaluations of 1,010 ceramic restorations, including inlays and onlays, placed in 299 patients during a 39-month period, that were reexamined at 9 to 12 years postplacement. PMID- 11913282 TI - Achieving high-level esthetics with CEREC. AB - This article describes the evolution of the CEREC CAD/CAM restorative system and its ability to create esthetic, durable ceramic- and resin-based restorations chairside. Included is discussion about specific techniques for color enhancement and modification to increase the esthetic result. The three machinable materials currently available for use with the CEREC system also are discussed. PMID- 11913283 TI - CEREC: the power of technology. AB - Dentistry has had a remarkable history. Technological advances have contributed immeasurably to efficient procedures, quality treatment, and patient satisfaction. CEREC, a computer-assisted design/computer-assisted manufacture (CAD/CAM) development that has been on the market for 15 years, is one such innovation. Capable of providing a durable, cosmetic, nonmetal filling in only one appointment and in less than an hour, the efficient use of the CEREC unit contributes markedly to quality care, patient satisfaction, and practice profit. CEREC, an acronym for ceramic-reconstruction, is a technology developed in 1985 by two Swiss researchers, a dentist and an electronics engineer, from the University of Zurich. With 15 years of research and development and more restorations placed than any comparable unit, the CEREC family of products has earned its role in dental history as the technology that gives patients one of the finest restorations in the world in only one visit. PMID- 11913284 TI - Esthetic restorations designed with confidence and predictability. AB - When the ProCAD product line is used in combination with the CEREC system, clinicians have at their disposal an armamentarium of materials with which to achieve greater predictability, precision, and control in fabricating chairside all-ceramic restorations. At a time when time is the dentist's and patient's most precious commodity, the ProCAD line enables clinicians to definitively restore form, function, and esthetics in a single visit, regardless of the specific individual indirect treatment indication (Figures 13A through 13C, and 14A through 14C). PMID- 11913285 TI - Properties and applications of a new composite block for CAD/CAM. PMID- 11913286 TI - The evolution of a chairside CAD/CAM system for dental restorations. AB - Since its introduction more than 15 years ago, the CEREC method for the chairside fabrication of all-ceramic restorations has steadily earned a loyal following among dentists. The rapid creation of tooth-colored restorations, including full coverage crowns, at chairside has been achieved through quick and easy optical impression-taking, simple designing, and precise machining. This article describes the evolution of this technology's development and its incorporation into the dental practice. PMID- 11913287 TI - The evolution and development of a chairside machinable ceramic material. PMID- 11913288 TI - CEREC: science, research, and clinical application. AB - Computer-aided design/computer-aided manufacture (CAD/CAM), as embodied in the CEREC system, enables the fabrication of high-quality esthetic ceramic restorations chairside. Resin cements are used to complete the clinical process, providing a high-strength, stress-free adhesive assembly. The current generation of CEREC technology can be used to create restorations with a repeatable 50 micron adhesive interface width. Systematic analysis of the literature on clinical trials of CEREC restorations indicates a high level of clinical success. CEREC technology is compatible with contemporary, conservative cavity preparation design and limits pulpal trauma by completing complex restorations in a single visit. The occlusal form of CEREC restorations can be machined with full regard to the patient's occlusion, using functionally generated pathway recordings. PMID- 11913289 TI - Effect of peroxide concentration and brushing on whitening clinical response. AB - This clinical trial compared the effects of hydrogen peroxide concentration and toothbrushing on clinical response to vital bleaching. Tooth bleaching was accomplished with a flexible, polyethylene strip coated with a hydrogen peroxide bleaching gel worn for 30 minutes twice daily over a 14-day period. A total of 36 subjects were randomized to 1 of 3 treatment groups: 5.3% hydrogen peroxide strip plus prebrushing, 6.5% hydrogen peroxide strip plus prebrushing, or 6.5% hydrogen peroxide strip without prebrushing. Two groups brushed with regular anticavity toothpaste immediately before bleaching, while the other group performed ad libitum brushing only. Tooth color was measured over a 14-day period using digital images of the anterior dentition. Over the 14-day treatment period, all 3 strip groups experienced highly significant (P < 0.001) whitening as evidenced by decreased yellowness (delta b*) and increased brightness (delta L*), as well as composite color change (delta E*) relative to baseline. Keeping brushing constant, the 6.5% hydrogen peroxide strip plus prebrushing group experienced a 31% to 60% improvement in whitening relative to the 5.3% hydrogen peroxide standard. Keeping concentration constant at 6.5% hydrogen peroxide, the prebrushing group experienced a directional 5% to 33% improvement in whitening relative to no prebrushing. All treatments were generally well tolerated. This study demonstrates that for strip-based delivery, increasing hydrogen peroxide concentration to 6.5% results in a significant improvement in efficacy with few tolerability trade-offs. PMID- 11913290 TI - Tooth whitening in children. AB - Although there are several case reports of vital tooth bleaching in children, there is limited clinical trial evidence of the safety or efficacy of this practice. Accordingly, a new clinical trial was conducted to evaluate the effects of 2 different bleaching systems, a 6.5% hydrogen peroxide strip system and a 10% carbamide peroxide tray system, in a population of preteens and teens. A total of 106 volunteers, aged 11 to 18 years, took part in this 8-week study. Patients were randomized by a ratio of 2:1 to the strip or tray groups, with each group treating the maxillary arch first and then the mandibular arch for 4 consecutive weeks each. Individuals assigned to the strip group used the system twice daily for 30 minutes (a total of 56 contact hours over the 8-week study). Those assigned to the tray group used that system overnight (approximately 448 contact hours). Digital images were obtained at baseline and after every 2-week treatment period. Average tooth color was determined in L*, a*, b* color space, where L* indicated lightness, a* indicated red-green, and b* indicated yellow-blue. Both systems significantly whitened teeth (P < 0.0001). While there were no significant differences between groups with respect to the primary whitening response (delta b*) on the maxillary teeth, 4 weeks of overnight treatment with the 10% carbamide peroxide tray (approximately 224 contact hours) yielded statistically significant whitening (P < 0.05) on the mandibular teeth compared with the 6.5% hydrogen peroxide strip used for 28 hours. Both tooth-whitening systems had similar sensitivity/irritation reported after instructed use. This research demonstrates that tooth whitening in teens may be safely accomplished using either the short-contact-time hydrogen peroxide bleaching strips or the overnight carbamide peroxide tray systems tested in this study. PMID- 11913291 TI - Daily use of whitening strips on tetracycline-stained teeth: comparative results after 2 months. AB - This article reviews the efficacy of a new 6.5% hydrogen peroxide tooth-whitening gel strip for bleaching teeth that have been intrinsically stained from tetracycline. Given the severity of staining in the cases presented during a recently conducted clinical trial, the resulting efficacy is dramatic. Additionally, the continuous use of these strips for 30 minutes per day, twice daily for 2 months with no meaningful adverse effects is noteworthy. PMID- 11913292 TI - Comparative clinical efficacy of two professional bleaching systems. AB - A randomized, parallel, examiner-blind clinical trial was conducted to compare two professionally dispensed vital bleaching systems. A total of 20 subjects were randomized to either the Crest Professional Whitestrips system or the Nite White Excel 2 tray system, balancing for baseline tooth color. During the 2-week treatment period, subjects were instructed to use the strip system for one-half hour twice daily (14 contact hours) and the tray system for 2 hours daily (28 contact hours). Only the maxillary teeth were treated, and all bleaching was unsupervised. Whitening was measured objectively by comparing digital images of the maxillary anterior dentition at pretreatment and posttreatment. Compared with baseline, both the strip and tray systems exhibited statistically significant decreases (P < 0.05) in yellowness (delta b*) and increases in brightness (delta L*). Compared with the tray-system group, the strip-system group experienced significant additional whitening (P < 0.03), as evidenced by a 62% to 209% improvement in tooth color (delta b*, delta L*, and delta E*) after 14 days. Both systems were generally well tolerated, and none of the subjects discontinued treatment early because of an adverse event. Under the conditions tested, this clinical trial demonstrates that the 14-contact-hour treatment with the strip system resulted in superior whitening efficacy compared with the 28-contact-hour treatment with the tray system. PMID- 11913293 TI - Whitening paradigms 1 year later: introduction of a novel professional tooth bleaching system. AB - The recent development of a novel "trayless" delivery system for vital bleaching shifted treatment paradigms and spurred surprising growth in tooth whitening. New research on a professional-only system (Crest Professional Whitestrips) further expands treatment options. This easy-to-use hydrogen peroxide strip system represents an effective and safe approach for increased whitening. Clinical applications are extensive, ranging from general, point-of-entry whitening to chronic use in difficult case types. PMID- 11913294 TI - Peroxide interactions with hard tissues: effects on surface hardness and surface/subsurface ultrastructural properties. AB - Laboratory studies were performed to assess the impact of peroxide bleaching on enamel surface and subsurface physical and ultrastructural properties. Human enamel blocks were prepared, polished, and measured for native color. Cyclic bleaching treatments were carried out with soaks in whole stimulated saliva interspersed with bleaching treatments using bulk bleaching gels from commercial bleaching systems including Opalescence (20% and 10% carbamide peroxide systems) and Crest Whitestrips, a hydrogen peroxide gel formula, at doses of 5.3% and 6.5% hydrogen peroxide. Treatments ranged from conditions of normal use (14 hours as recommended for Crest Whitestrips) to excessive bleaching (70 hours). Controls included nontreated as well as treatments with placebo (not containing peroxide) gels. Surface hardness and confocal laser scanning microscopy (CLSM) techniques were used to characterize the effects of bleaching on the physical properties and ultrastructure of the teeth. Tooth color measurements revealed dose-response bleaching in vitro with the increases in L* and decreases in b* normally expected with effective bleaching. Placebo control treatments did not bleach. Surface hardness measurements showed no decreases associated with tooth bleaching. CLSM measurements also showed no effects from tooth bleaches on the surface or subsurface prism architecture of enamel. This was opposed to significant changes seen with even moderate levels of demineralization associated with the caries process. These studies support: (1) the safety of Crest Whitestrips formulas for enamel surfaces and tooth subsurfaces; and (2) the generic safety of peroxide bleaching of hard tissues associated with conditions of both recommended use and overuse. PMID- 11913295 TI - Overview of a professional tooth-whitening system containing 6.5% hydrogen peroxide whitening strips. AB - Professionally dispensed, at-home tooth whitening began with 10% carbamide peroxide gels applied to the dentition with custom-made trays. In the 1990s, higher-concentration carbamide peroxide gels were introduced to achieve faster results. Today, 15% and 20% carbamide peroxide gels are commonly used. Recently, a new vital tooth-whitening technique that uses a flexible strip rather than a tray to apply a 5.3% hydrogen peroxide whitening gel was introduced. The new strip-based product was shown to provide whitening equivalent to a 10% carbamide peroxide tray with half the wear time. In addition, the strip eliminated the need to custom fabricate trays for each patient. This article provides an overview of a professionally distributed strip-based whitening system and reviews some of the clinical data which supports the efficacy of the product. This new whitening system includes 42 mandibular and 42 maxillary strips at a higher concentration of 6.5% hydrogen peroxide. In addition, the system also includes a novel dual action whitening dentifrice to prevent future staining postbleaching and an extrasoft toothbrush. Clinically, the professionally distributed strip-based whitening system provided 96% more efficacy than a popular carbamide plus hydrogen peroxide (equivalent to 10% carbamide peroxide) tray system and 52% more whitening than the 5.3% hydrogen peroxide strip system. PMID- 11913296 TI - Training: the new frontier. PMID- 11913297 TI - Microdentistry: current pit-and-fissure caries management. AB - The systemic application of fluoride has rendered enamel more resistant to decalcification and decay. While the incidence and severity of dental caries has decreased, the diagnosis of caries in this enamel environment has created significant new challenges and dilemmas. Traditional diagnostic methodology and treatment philosophy no longer seem appropriate. This article describes the historical perspectives, current diagnostic challenges, and the treatment dilemmas of occlusal pit-and-fissure systems currently facing the dental profession. PMID- 11913298 TI - A forced rapid extrusion technique for compromised teeth. PMID- 11913299 TI - Clinic jackets and gowns: a misunderstood personal protection equipment requirement? PMID- 11913300 TI - Total mandibular reconstruction with cancellous cellular bone graft, allogeneic bone crib, and endosseous implants. AB - It is not often that the oral and maxillofacial surgeon is faced with the challenge of restoring a defect that involves the entire mandible. The case presented in this article illustrates the management of such a case. Trauma, infection, neoplasm, and congenital malformations can lead to a discontinuous, deficient, or even absent mandible. In planning the reconstruction of a discontinuity of the mandible, the surgeon may need to manage one or more problems, including loss of adequate denture-bearing bone, loss of speech articulation, loss of control of the tongue and masticatory function, xerostomia as a result of the loss of sublingual and submandibular glands, loss of an intact deglutition mechanism, and a loss of facial form. PMID- 11913301 TI - The management of gingival and esthetic problems: a clinical case. AB - The coordinated management of gingival and esthetic problems is important in dentistry. In addition, the most difficult variable to control in the treatment of anterior esthetics is the soft tissue. This article discusses the relationship between the surrounding soft tissue and the emerging crown form. The authors introduce a soft-tissue index model technique that will help the technician in fabricating the final crown with a form similar to the provisional. In this case report, provisional crowns are fabricated with enough tissue support to provide for an optimal crown-gingival interface. The present case demonstrates how to manage the gingival margin in the provisional stage and how to transfer the provisional and soft tissue to the laboratory model. PMID- 11913302 TI - Space requirements for implant-retained bar-and-clip overdentures. PMID- 11913303 TI - Locating the centric relation prematurity with a computerized occlusal analysis system. AB - Locating the first tooth contact that interferes with freedom of movement in and out of centric relation has been the diagnostic and treatment objective of most occlusal therapies. The centric relation prematurity can be located by various methods, which involve operator-guided mandibular positioning combined with the patient's subjective assessment of his or her perceived first tooth contact. The method known as bimanual manipulation has been widely recognized and accepted as a predictable method of determining and verifying the centric relation position. The first occlusal contact that results when the mandible is closed on a correct centric relation axis is known as the centric relation prematurity. An alternative procedure combines bimanual manipulation with the simultaneous recording of the sequence of resultant tooth contacts using a computerized occlusal analysis system. This alternative offers a significant improvement in the precision of locating the first tooth contact. This article describes a method of identifying the first tooth contact while not relying on the patient's subjective assessment of his or her perceived occlusal feel. PMID- 11913304 TI - An evaluation of a commercial chewing gum in combination with normal toothbrushing for reducing dental plaque and gingivitis. AB - New evidence suggests a beneficial outcome to chewing a sugarless gum as an added component to a regular, twice-daily toothbrushing regimen. Results of a 4-week study performed on 78 adults with preexisting gingivitis showed a significant reduction of dental plaque and gingivitis when the test group of 39 adults chewed 2 pieces of ARM & HAMMER Dental Care The Baking Soda Gum (AHDC)--a sugar-free chewing gum containing sorbitol, malitol, xylitol, and sodium bicarbonate--for 20 minutes twice a day in conjunction with once-daily toothbrushing for 60 seconds. The control group, also comprised of 39 adults, used breath mints (the study placebo) twice a day in conjunction with the same toothbrushing regimen. There were no statistically significant differences in plaque and gingivitis scores at the baseline examination. Using the Quigley-Hein Plaque Index, the test group experienced a 17% reduction in plaque over 4 weeks, while the control group reduced their plaque amounts by approximately 9% over the same period. Lobene's Mean Gingivitis Index scores were equally significant: a nearly 10% decline for the test group compared to almost 2% for the control group. This article describes the 4-week study and its promising results. PMID- 11913305 TI - A graphic display for the presentation of site-wise odds ratios for score transitions to augment the traditional findings from clinical studies employing dental indexes. AB - Many of the parameters used in clinical dental research involve the assessment of a condition at each of a number of sites within the mouth. Traditionally, such measurements are averaged over all sites within the mouth (or over all sites of a specified type) for each study participant before statistical analysis. However, a consideration of the original, site-wise scores may provide some additional insights into the performance of therapeutic modalities that might not be made evident through an application of the traditional, means-based approach. A method based on the calculation of site-wise odds ratios of certain types of baseline-to final examination score transitions was applied to the modified gingival index data from two clinical studies performed to investigate the effect of the daily chewing of a commercially available chewing gum relative to a mint control. A graphical display of these site-specific findings was prepared, which indicated that the chewing gum regimen tended to be associated with a higher frequency of occurrence of favorable score transitions than was the mint control at several measurement sites throughout the mouth. PMID- 11913306 TI - Effects of a Baking Soda Gum on extrinsic dental stain: results of a longitudinal 4-week assessment. AB - An evaluation of the effects of ARM & HAMMER DENTAL CARE The Baking Soda Gum (AHDC) on extrinsic dental stain was made in 48 subjects presenting with measurable extrinsic stain. The subjects were randomized to use either the baking soda gum or a non-baking soda placebo gum for 20 minutes twice daily after lunch and dinner while brushing once daily. The procedure of limited brushing was chosen to simulate the level of hygiene normally practiced by participants entering a clinical study. After 4 weeks, the reduction in measurable extrinsic stain in the baking soda gum group was statistically significant (P = .0044) relative to baseline. Statistical analysis of the placebo gum group revealed no significant change in extrinsic stain from baseline. The magnitude of the unadjusted longitudinal reduction in extrinsic stain in the baking soda gum group was 29.7% at 4 weeks. PMID- 11913307 TI - Efficacy of baking soda-containing chewing gum in removing natural tooth stain. AB - A 14-week, double-blind, randomized clinical trial was conducted with 126 healthy volunteers to compare the efficacy of twice-daily use of 3 baking soda-containing chewing gums in removing natural tooth stain when used in conjunction with a program of regular oral hygiene. All 3 chewing gums significantly reduced extrinsic stain (P < .0001) and improved the whitened appearance of teeth (P < .0001) at both the 2-week interim and the final 4-week evaluations. ARM & HAMMER DENTAL CARE The Baking Soda Gum (AHDC) reduced dental stain by 70.8%, compared to reductions of 71.9% and 65.3%, after use of 2 experimental gum formulations. Whitened appearance improved by 1.73 shade tabs using AHDC gum, and up to 2.49 shade tabs with the experimental formulations. These results suggest that the use of baking soda-containing gum after meals, in conjunction with good oral hygiene, can improve both extrinsic dental staining and the whitened appearance of teeth. PMID- 11913308 TI - A clinical investigation to evaluate reduction in dental stain provided by the once-daily use of a breath mint or chewing gum. AB - An evaluation of the effects of ARM & HAMMER DENTAL CARE The Baking Soda Gum on extrinsic dental stain was performed on 85 subjects presenting with measurable extrinsic stain. The subjects were randomized to use either the baking soda chewing gum or a breath mint placebo once daily after lunch while brushing once daily. The chewing gum was chewed for 20 minutes for each use and the breath mint was kept in the mouth until completely dissolved. The subjects were instructed to brush once daily to simulate the level of hygiene normally practiced by subjects, thereby avoiding the well-reported Hawthorne effect experienced in clinical trials instituting twice-daily brushing. Examinations postbaseline were performed after 2 and 4 weeks. The reduction in measurable extrinsic stain in the baking soda gum group vs the breath mint control was statistically significant at 2 weeks (P < .0002) and at 4 weeks (P < .0008). Statistical analysis of the data revealed a significant change in extrinsic stain from baseline for both groups. The magnitude of the unadjusted longitudinal reduction in extrinsic stain in the baking soda gum group was 51% at 4 weeks. PMID- 11913309 TI - An evaluation of sodium bicarbonate chewing gum as a supplement to toothbrushing for removal of dental plaque from children's teeth. AB - The purpose of this human clinical study was to determine whether a commercial chewing gum containing 5% sodium bicarbonate (ARM & HAMMER DENTAL CARE The Baking Soda Gum [AHDC]) was effective in removing dental plaque when used as a supplement to regular toothbrushing by children. Healthy children (N = 28, average age = 11 years) were randomly distributed into 2 groups. One group was instructed to chew 2 tablets of AHDC chewing gum for 20 minutes 2 times each day (after lunch and dinner) in addition to their normal toothbrushing regimen. The other group used a sugarless mint tablet twice daily during the same period in addition to toothbrushing. After 1 week of using their assigned product, all participants were again examined for oral health and plaque. After a 1-week washout period, subjects were crossed over to the opposite group. Among the 21 participants completing the study, the AHDC chewing gum significantly (P < .0001) reduced plaque by 15% after 1 week compared to the mint tablet control, as measured by the Modified Quigley-Hein Plaque Index. When longitudinally compared to the baseline plaque scores, the gum resulted in a significant (P < .01) 10% reduction of plaque on the teeth. Subanalysis of the data showed that the AHDC chewing gum was particularly effective on the lingual surfaces and the posterior teeth and least effective on the facial surfaces of the anterior teeth, which do not readily come into direct contact with the gum during mastication. The bicarbonate gum demonstrated significant plaque reduction in all other areas of the mouth, even on tooth surfaces not directly contacted during chewing. Compliance with the chewing gum regimen was excellent, and oral health exams did not indicate any adverse events among children using either the chewing gum or mint tablets. In this study, regular use of AHDC chewing gum was safe and effective in removing dental plaque and served as a significant complement to the daily toothbrushing regimen of children. PMID- 11913310 TI - An evaluation of sodium bicarbonate chewing gum in reducing dental plaque and gingivitis in conjunction with regular toothbrushing. AB - The purpose of the present study was to determine the ability of a chewing gum containing 5% sodium bicarbonate to remove dental plaque and reduce gingivitis when used as a supplement to daily toothbrushing. The study group consisted of 88 adults with moderate gingivitis. Participants were divided into 4 groups and instructed to chew the study gum 0 (control), 1, 2, or 3 times daily for 1 month in addition to regular daily toothbrushing. Chewing sodium bicarbonate-containing gum significantly (P < .05) reduced plaque after 1 week, with progressively greater reductions occurring after 2 and 4 weeks of gum chewing. No correlation was observed between plaque reduction and the number of times per day that the gum was chewed. By week 4, plaque reduction of approximately 16% was achieved in all groups using the gum as compared with the control group. Reductions in gingivitis were observed in all participants who chewed the gum, and these reductions achieved statistical significance by week 4. Slightly greater improvements in gingivitis were achieved in participants who chewed the gum 2 or 3 times daily as compared with the control group and those who chewed the gum once daily. No adverse effects on the oral tissues were observed in any of the participants for the duration of the study. In conclusion, regular use of a chewing gum containing 5% sodium bicarbonate appears safe and effective for the removal of dental plaque and reduction of gingivitis when used in conjunction with daily toothbrushing. PMID- 11913311 TI - Oral malodor control afforded through the use of sodium bicarbonate-containing chewing gum. AB - Published research indicates that regular use of toothpaste containing sodium bicarbonate is effective in reducing volatile sulfur compounds (VSCs) and oral malodor. Gaffar initially reported on the use of sodium bicarbonate as an agent to affect VSCs, indicating that sodium bicarbonate had the potential to alter the VSCs to nonvolatile compounds. Chewing gum would also be suspected of providing benefits in controlling oral malodor through its claimed ability to mechanically aid in the removal of dental plaque. Based on the length of chewing time, the opportunity exists for chewing gum to reach places in the mouth that might be missed during brushing. This activity could contribute to reductions in the amount of viable plaque mass that could produce VSCs. PMID- 11913312 TI - A simplified impression technique for dental implants. AB - Dental implants have been considered an acceptable form of dental treatment since the early 1980s. A number of studies have been published describing impression techniques for dental implants. Many of the techniques described are so complex that they may seem daunting to the average restorative dentist. Most general practitioners do not wish to attempt to restore dental implants. This article describes a very simple, yet extremely accurate, technique for making impressions of dental implant fixtures. PMID- 11913313 TI - Illustrating predictable anterior and posterior esthetic results: two case studies. AB - More natural-looking restorations are possible today because of advances in the quality of impression materials and the techniques used for tooth preparation and cementation. This article presents two cases that illustrate the use of a new generation of impression materials, bonded resin cement, and pressable porcelain to obtain the desired esthetic result. PMID- 11913314 TI - Relining extension-base removable partial dentures. AB - Modern prosthodontic therapy must continue to address the inevitable consequence of tooth loss--that is, alveolar process atrophy. The use of removable partial dentures (RPDs) to treat partial edentia must rely, in part, on support from the ever-deteriorating edentulous ridges. Extension-base RPDs are particularly affected as the tissue-base relationship changes over time after extraction of the natural teeth. This article reviews the concept of observed changes in the residual edentulous alveolar ridge and conventional procedures used to compensate for this bone loss. Finally, a practical procedure is presented which uses convenient and accurate materials that can be readily incorporated into the modern dental practice. PMID- 11913315 TI - Success with inlays/onlays: the seven essentials. AB - Direct restorative materials have revolutionized the industry over the last several years. They offer the convenience to the patient of a one-visit procedure and they decrease overhead in the dental practice. However, it is essential to remember that direct procedures are not always the treatment of choice. For this reason, we should not discount the need for--or the value of--indirect techniques and laboratory-based procedures, such as inlays and onlays. Fit, form, and function can be more predictable when a procedure involves the ultraprecise workings of the dental laboratory--if the impression-making and other focus areas are given their due significance. This article provides key information about and clinical tips for the seven main facets of inlay/onlay fabrications that will lead to clinical success: diagnosis, tooth preparation, tissue management, impression making, temporization, laboratory communications, and final fit/occlusion. PMID- 11913316 TI - Predictable fixed prosthodontics: technique is the key to success. AB - This article reviews step-by-step clinical procedures for fixed prosthodontics that include a discussion of the following case parameters: (1) proper diagnosis and case selection; (2) tooth preparation; (3) soft tissue management; (4) impression making; (5) provisionalization; (6) additional laboratory diagnostic information; (7) case try in; and (8) case delivery. When the clinical principles of these procedures are faithfully followed, the success and predictability of any restorative endeavor will be greatly enhanced. PMID- 11913317 TI - Opportunities in the practice. AB - These are just a few of the changes that took place in the overall scheduling changes engineered for this dentist. The result was a 32.7% increase in practice production in less than 4 months, evidence clearly showing that the practice originally had significant unrealized potential. Unfortunately, the systems originally being used led the practice to bump up against a ceiling that appeared to have no more capacity. By re-engineering the schedule and addressing other systems, significant growth was experienced. At the end of 12 months, the practice had increased total production by more than 40%. Equally important, the dentist reported that he was far less stressed and was actually able to enjoy his practice once again. The main point of this article is to help dentists and teams understand why they hit a ceiling and are unable to squeeze more out of their practice. Each additional attempt to increase productivity creates significant stress and eventually leads to customer service problems. By re-engineering the systems to accommodate both the current and future potential of the practice, significant profit and growth can be achieved, while experiencing much greater daily satisfaction along with a significant reduction of stress. PMID- 11913318 TI - Esthetic implant dentistry. AB - Consistency of results, reliability of treatment modalities, and long-term prognosis require scientific approaches to therapeutic procedures. Reliable and unbiased information rooted in academics and research assist the health care professional use current technology to develop the best course of action to restore or enhance the best esthetic outcome for the patient. Without anticipating potential failure, any immediate success is limited to initial satisfaction while ignoring the far greater elements of a future problematic outcome. This article proposes procedures for soft and hard tissue preservation, repair, or reconstruction in esthetic implant dentistry and emphasizes that an essential prerequisite to effective esthetic therapy is the establishment of a complete and accurate diagnosis. PMID- 11913319 TI - Tissue preservation for single-tooth anterior esthetics. AB - Placing implants in the esthetic zone can be difficult because of the expectations of the patient. Inevitably, the hard and soft tissue architecture is lost after extraction in the traditional two-stage procedure, and additional surgical procedures are required to restore the tissue that has been lost. The authors describe techniques that can be applied to retain ideal soft tissue if it is present and predictably restore tissue that is less than ideal. PMID- 11913320 TI - Composite materials in restorative dentistry. Interview by Mark J. Friedman. PMID- 11913321 TI - The esthetic effects of implant platform selection. AB - Implant dentistry has undergone considerable evolution over the history of modern endosseous implant therapy. With the advent of enhanced bone and soft tissue manipulation procedures, improved implant hardware components, and enough experience for the procedures to evolve, an implant restoration today may be indistinguishable from a natural tooth. Guidelines are discussed for implant selection and implant placement dictated by the desired functional and esthetic result. The implant platform should be selected only after the tooth and soft tissue dimensions are decided. In addition, the largest platform diameter that fits within the normal root contours should be selected. Surgically, that platform should be placed 3 mm apically and 1 mm lingually to the buccal emergence point. The buccal cementoenamel junction or free gingival margin thus becomes the key determinant for implant position. Specific implant dimensions are discussed for each tooth to provide appropriate guidelines for the practitioner. PMID- 11913322 TI - Perspectives on esthetics in implant dentistry. AB - Esthetic outcomes are related more to soft tissue volume and adaptation than to implant components. A preoperative understanding of the influence and inter relationship of bone volume, soft tissue volume and contours, attainable augmentation goals, and the achievable realistic restorative results is important. This article shows the application of routine components in methods to solve routine commonly encountered clinical problems, including matching a missing central incisor to the adjacent dentition and soft tissue, esthetic provisionalization for the hopeless anterior tooth after extraction, and elective replacement of an upper denture with a fixed implant restoration without the use of bone grafting. PMID- 11913323 TI - Periodontal splinting with a thin high-modulus polyethylene ribbon. PMID- 11913325 TI - Adolescent tobacco use: the protective effects of developmental assets. AB - PURPOSE: To examine the effects of 10 youth developmental assets on adolescent tobacco use. DESIGN: Survey of a randomly selected sample using in-home interviewing methodology. SETTING: Inner-city areas of two midsized Midwestern cities. SUBJECTS: The researchers studied 1,350 teen-parent pairs. MEASURES: Demographic information, adolescent self-reported tobacco use, eight developmental asset Likert scales, and two one-item developmental asset measures. RESULTS: The response rate was 51%. Logistic regression results indicate that youth who possess nine of 10 developmental assets examined are significantly less likely to report tobacco use than youth with low levels of assets. Adjusting for youth age, race, gender, parental income and education, and family structure, significant odds ratios include the following: nonparental adult role model, 2.09 (95% confidence interval [CI] = 1.45, 3.02); peer role models, 2.48 (95% CI = 1.87, 3.29); family communication, 1.73 (95% CI = 1.29, 2.31); use of time (organized groups), 1.77 (95% CI = 1.28, 2.44); use of time (religion), 2.49 (95% CI = 1.86, 3.33); good health practices (exercise/nutrition), 1.61 (95% CI = 1.21, 2.14); community involvement, 1.66 (95% CI = 1.07, 2.58); future aspirations, 2.06 (95% CI = 1.42, 2.99); and responsible choices, 2.21 (95% CI = 1.55, 3.15). CONCLUSIONS: The findings of this study support the view that certain developmental assets may serve to protect youth from risk-taking behaviors, particularly tobacco use. Limitations include cross-sectional data and three scales with alphas below .7. PMID- 11913324 TI - Applying theory of planned behavior to fruit and vegetable consumption of young adolescents. AB - PURPOSE: The purpose of this research was to investigate how well Theory of Planned Behavior (TPB) predicted frequency of consumption of fruits and vegetables among seventh graders and to investigate whether gender or socioeconomic status (SES) had moderating effects on the relationships between the constructs in the model. DESIGN: Two identical self-administered paper and pencil surveys were distributed 6 months apart. Path analyses of the TPB models by gender and SES were performed with AMOS software. SETTING: The study occurred in eight middle schools from school districts in the Minneapolis--St. Paul metropolitan area, where a minimum of 20% of the students qualified for free or reduced-price meals. SUBJECTS: Of the invited seventh graders, 1406 (74%) completed data for the model constructs at baseline and for behavior at interim data collection. MEASURES: We assessed frequency of consumption of fruits and vegetables and attitudes, subjective norms, barriers, and intentions related to this behavior. Three levels of SES were created based on questions about free lunch, parents' education, and work. RESULTS: No significant direct effects of attitudes and subjective norms on behavior were found, but for barriers, both indirect effects through intentions and direct effects on behavior were significant. Seven percent of the variation in the frequency of fruit and vegetable consumption and 31% of the variation in intention to eat more fruits and vegetables were explained by the model. Gender appeared to have moderating effects on the relationships between attitudes and intention and between intentions and behavior in the model. There were no moderator effects of SES, but the larger dropout rates from the low-SES group could have caused the groups to become more homogeneous. CONCLUSION: The data fit the model well; still, large proportions of the variance in the frequency of consumption of fruits and vegetables were unexplained. This may be related to the operationalization of the model constructs or to missing important factors in the model. Furthermore, there appeared to be strong relationships between subjective norms or barriers and intentions as well as gender differences in the strengths of the relationships in the model that may have implications for intervention methods or messages. PMID- 11913326 TI - Health and behavior: the interplay of biological, behavioral, and social influences: summary of an Institute of Medicine report. AB - An Institute of Medicine committee was convened to explore the links between biological, psychosocial, and behavioral factors and health and to review effective applications of behavioral interventions. Based on the evidence about interactions of the physiological responses to stress, behavioral choices, and social influences, the committee encouraged additional research efforts to explore the integration of these variables and to evaluate their mechanisms. An understanding of the social factors influencing behavior is growing and should be considered in programs and policies for public health, in addition to individual behavior and physiological status. Interventions to change behaviors have been directed toward individuals, communities, and society. Many intervention trials have documented the capacity of interventions to modify risk factors. However, more trials that include measures of morbidity and mortality to determine if the strategy has the desired health effects are needed. Behavior can be changed and new behaviors can be taught. Maintaining behavior changes is a greater challenge. Although short-term changes in behavior following interventions are encouraging, long-duration efforts are needed to improve health outcomes and to provide long term assessments of effectiveness. Interventions aimed at any level can influence behavior change; however, existing research suggests that concurrent interventions at multiple levels are most likely to sustain behavior change and should be encouraged. PMID- 11913327 TI - Health promotion for people with physical, cognitive and sensory disabilities: an emerging national priority. AB - Despite the growth in health promotion programs for able-bodied people, very little effort has been devoted to developing programs for people with physical and cognitive disabilities. Programs for people with disabilities must be developed with full recognition of limitations caused by both the primary and secondary disability. PMID- 11913328 TI - Differences in health care costs and utilization among adults with selected lifestyle-related risk factors. AB - PURPOSE: This study examined the relationship between lifestyle-related health risks and health care costs and utilization in adults. DESIGN: A 2-year prospective study with no intervention was used to compare health care utilization and costs in employees with different levels of health risks. SETTING: Data were collected at a primarily white-collar worksite during 1994 and 1995. SUBJECTS: Subjects included 982 employees and spouses, mean age 32.1 +/- 10.1 years. MEASURES: Employee medical claims obtained from a third-party administrator were analyzed with respect to health care expenses and utilization. Exercise habits, stress, and overall wellness were assessed by self-report and obesity by the body mass index (BMI). Regression, regression with outliers removed, and odds ratios were used to analyze the associations. RESULTS: Employees who were at high risk for overall wellness (2.4 times), stress (1.9 times), and obesity (1.7 times) were more likely to have high health care costs (> $5,000) than subjects not at high risk. Mean total medical costs also were greater for high-risk subjects compared to lower risk subjects for overall wellness (difference = $1,973; F = 10.65, p = .001), stress (difference = $1,137; F = 7.35, p = .007), and obesity (difference = $1,092; F = 9.09, p = .003). The exercise habits measure was not significantly associated with health care costs or utilization. CONCLUSIONS: Results indicate that health risks, particularly obesity, stress, and general lifestyle, are significant predictors of health care costs and utilization in employed young adults. PMID- 11913329 TI - Transforming the vision into reality. PMID- 11913330 TI - Testosterone and nonverbal intelligence in right-handed men with successful and unsuccessful educational levels. AB - The relationship between serum total testosterone (T) concentration and fluid intelligence (nonverbal, spatial) was studied in consistently right-handed men with successful (S) or unsuccessful educational levels (NS). Hand preference was assessed by the Edinburgh Handedness Inventory. Nonverbal intelligence was measured by Cattell's Culture Fair Intelligence Test. Serum T level was determined using chemiluminescence enzyme-immunoassay on hormone autoanalyzer. There was no significant difference between the mean T levels of the S subjects and NS subjects, although S-men tended to have higher T levels than NS-men. The mean IQ was found to be significantly higher in S-men than NS-men. In the total sample (S + NS men), the correlation between T to IQ was best described by a polynomial regression (3rd order), exhibiting an inverse U-shaped regression. In S-men, the relationship between T and IQ was best described by a polynomial regression equation of the 3rd order; however, the relationship was not U-shaped, but rather a positive correlation (low T: low IQ and high T high IQ). In NS-men, there was an inverse U-shaped correlation between T and IQ (low and very high T: low IQ and moderate T: high IQ). The present data suggest that (i) very low and very high serum T concentrations may be disadvantageous, (ii) moderate T levels may be advantageous for general fluid intelligence, and (iii) a prewired cerebral organization may be essential for the T effects on cognitive abilities. PMID- 11913331 TI - Demographic effects on the Trail Making Test in narcotic/other opiate abusers. AB - Demographic effects on the Trail Making Test (TMT), a test often used to screen for cognitive impairments, were examined in a sample of narcotic/other opiate abusers in drug abuse treatment programs. A sample was drawn from the Drug Abuse Treatment Outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991 through 1993 in 96 programs within 11 cities in the United States. The number of narcotic/other opiate abusers' scores available for analysis was 191. Data were analyzed to determine the effects of sex, ethnicity, age, and education on the two parts of the TMT in this sample of narcotic/other opiate abusers. The variables of age and education level were statistically significantly related to TMT parts A and B, and ethnicity was statistically significant for part B of the TMT. R-square values for overall models were moderate (A = .34, B = .24), suggesting that demographic effects on the TMT are moderate. PMID- 11913332 TI - Do early onset conduct disordered adolescents perform like brain injured or normal adolescents on cognitive tests? AB - In order to better understand whether adolescents with early onset conduct disorder are predisposed to this disorder because of neurological problems, the present study compared the performance of early onset conduct disordered adolescents to adolescents with left hemisphere and right hemisphere brain injuries and normal controls. It was hypothesized that adolescents with early onset conduct disorder would perform similar to adolescents with left hemisphere injury, confirming theories that neurological dysfunction may predispose children to the development of conduct disorder. Fifteen adolescents with conduct disorder were compared on a battery of cognitive tests to 12 left hemisphere brain injured, 11 right hemisphere brain injured, and 15 normal middle school adolescents. F-tests indicated that there were significant differences among the four groups on all measures (p < .01). According to a series of t-tests on each of the nine cognitive measures, there were no differences found between the early onset conduct disordered subjects and the left hemisphere subjects. The early onset conduct disordered group performed worse than the right hemisphere group on 7 of the 9 comparisons, and worse than the normals on 9 of 9 comparisons. The normal group performed better than all three of the other groups. These results demonstrate that the pattern of neuropsychological performance by early onset conduct disordered adolescents was similar to that of left hemisphere injured adolescents and different from that of the right hemisphere injured and normal adolescents. Potential applications of this research include describing new approaches to treatment of this disorder based on their similarity with the left hemisphere brain injured group. PMID- 11913333 TI - Effect of risperidone on serum cytokines. AB - A variety of cytokines are dysregulated in schizophrenia, and some antipsychotic drugs effect cytokines. In order to examine the effect of risperidone on plasma cytokines, we measured the serum level of IL-1b, IL-2, IL-6, IL-12, and INF-g during acute states of illness, and after 4 weeks of treatment with risperidone in 19 schizophrenic patients. The patients' psychopathology was assessed by PANSS. Plasma IL-12 levels increased significantly after 4 weeks of treatment (p = .002). Plasma IL-b, IL-2, IL-6, and INF-g levels were not significantly different before and after treatment. There were no significant correlations between the changes in cytokine levels and the changes in PANSS scores. Increased IL-12 may contribute to activation of immune responses during treatment with risperidone. IL-12 may play an important role in immune responses related to neuropsychiatry. PMID- 11913334 TI - Derived trail making test indices in a sample of substance abusers: demographic effects. AB - Derived indices on the Trail Making Test (TMT), a test often used to screen for cognitive impairments, were examined in a sample of substance abusers in drug abuse treatment programs. A mixed race sample was drawn from electronic files of data from the Drug Abuse Treatment Outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991 through 1993 within 96 programs in 11 cities in the United States. Data were analyzed to determine the effects of demographic variables on derived indices created by adding, subtracting, multiplying, and dividing parts A and B of the TMT in this large treatment sample of substance abusers. The variables of sex, age, ethnicity, and education were statistically significant for selected derived indices of the TMT. PMID- 11913335 TI - Effects of maternal bilateral adrenalectomy on fetal rat cerebral cortex. AB - We investigated the effects of maternal bilateral adrenalectomy (ADX) on fetal rat cerebral cortices by assessing morphological and morphometrical parameters in light and electron microscopy studies. Pregnant adult rats underwent ADX on day 4 of gestation. Embryonic brain tissue from control and ADX groups were examined at each of the 10 embryonic stages, days 11 through 20 (E11-E20). Control and adrenalectomized pregnant females were sacrificed at each fetal stage, fetuses were removed from the uterine horns, and their brain tissue was processed for light and electron microscopy examination. In the ADX fetuses, the cortical laminae were thicker and overall cortex thickness was greater, but structural differentiation in the primitive cortical layers was delayed compared with that observed in controls. Apart from the differences in embryonic cell development noted at the structural level, we found no noticeable ultrastructural differences between the cerebral cortices of ADX and control fetuses at any of the stages studied. Overall, the ADX group's cortical tissue exhibited a greater degree of cell migration, an extended proliferative period in the cortical layers with the highest proportion of mitotic activity, and a greater thickness than that of control specimens. We believe that by eliminating the inhibitor effects of glucocorticoids, ADX effectively induces mitotic activity and extends the proliferative periods in developing cerebral cortex. PMID- 11913336 TI - Further manipulation of the stop-signal task: developmental changes in the ability to inhibit responding with longer stop-signal delays. AB - The stop-signal task, a measure of inhibitory control, was further modified in order to examine its suitability as a task for very young children. A previous study (Carver et al., 2001) showed that it can be successfully adapted for use with primary school-aged children. The present study manipulated the presentation of the signal to inhibit responding and found that this improved the likelihood of responding. A pre-primary school group of children (< 5 years, 6 months), a young primary school group (5 years, 7 months to 7 years, 6 months), and a mid primary school group (7 years, 7 months to 9 years, 6 months) participated in the study. The results emphasize the pre- and early school years as a sensitive time for the development of inhibitory skills. Measures of inhibitory control must therefore be age-appropriate and sensitive to these early developmental changes. PMID- 11913337 TI - Unique case of eleven Bell's palsy episodes. AB - Bell's palsy (BP) is a peripheral facial nerve paralysis of unknown etiology. It is not a life-threatening condition; however, incomplete recovery may leave an individual stigmatized functionally, occupationally as well as socially. Recurrent paralyses are seldom, noted in 7-8% of all BP cases. More than two BP relapses are even less frequent. Adour et al. (1977) reported only two patients with four BP episodes from 1700 patients. Only one patient with more than four BP recurrences in the group containing 2414 BP cases were reported by Yanagihara et al. (1984). The highest reported number of BP recurrences in the accessible literature has been nine. We are presenting an unusual patient who suffered a total of eleven relapses of an idiopathic facial nerve palsy. Description of the case along with review of the relevant literature are discussed. PMID- 11913338 TI - Human verbal working memory impairments associated with thalamic damage. AB - Although animal studies, human neuroimaging studies, and numerous theoretical models suggest possible contributions of the thalamus to working memory, there are very few reported deficits in human working memory following thalamic lesions. The present study examined working memory performance in six individuals with isolated thalamic stroke and found evidence of impairment on a number of working memory span tasks, but not on a forward digit-span measure. Examination of additional aspects of working memory performance (e.g., spatial and object working memory), analysis of subjects with other sites of thalamic stroke, and functional neuroimaging suggest a role of the thalamus in working memory. PMID- 11913339 TI - Side bias and accidents in Japan and India. AB - The purpose of this study was to examine (a) the incidence of leftward bias for paired organs (hand, foot, eye, and ear) in India (n = 418) and Japan (n = 697), and (b) the incidence of accident amongst individuals with leftward, rightward, and no bias across countries. The impetus for the study was obtained from a speculation that individuals with leftward bias commit more accidents than their counterparts. Data were collected with the help of a questionnaire on side bias, along with a check-list on accidents (included in the questionnaire). Results showed that participants from these two countries did not differ in terms of leftwardness for any of these forms of side bias. Left-handers were found to commit more accidents in India but not in Japan. Reanalysis of data in terms of left-, mixed-, and right-handedness indicated that mixed handers committed more accidents than extreme (left- + right-) handers in Japan but not in India. PMID- 11913340 TI - [Stanislav Konstantinovich Evtushenko (on his 60th birthday)]. PMID- 11913341 TI - [Characteristics of Neisseria meningitidis strains isolated from patients with symptoms of meningococcal meningitis in Poland in 1995-2000]. AB - Phenotype and genotype identification of 179 Neisseria meningitidis strains isolated from cerebrospinal fluid or blood of patients with meningococcal infection, hospitalized in Poland, was performed. This is the first analysis of that type conducted in Poland. We analyzed strains collected in 1995-2000 from laboratories located all over the country. Phenotype Neisseria meningitidis B:22:P1.14 was the predominant among analyzed invasive strains in Poland. Type 22 is characteristic to most of the strains isolated in our country. No strain from analyzed group belonged to known epidemic clusters. One penicillin resistant strain (MIC = 2 mg/l) and about 27% strains with decreased susceptibility to penicillin (0.1 = < MIC < 1.0 mg/l) were present among 166 N. meningitidis tested. All strains were susceptible to ciprofloxacin and rifampicin. PMID- 11913343 TI - Implications of human-animal interactions and bonds in the laboratory. PMID- 11913344 TI - [The restoration of endodontically treated teeth. A literature study of the mechanical behavior of post-core restorations]. AB - Rapid developments in dental health and science have resulted in an increasing number of teeth receiving post and core restoration. The dentist can choose from a wide range of restoration-modalities. Success or failure of these restorations largely depends on their ability to meet functional demands. This article presents a review of the literature on the mechanical behaviour of post-core restorations, as it is determined by functional forces, shape of the tooth and restoration and properties of the materials the construction encorporates. Some attention is drawn to the importance of failure characteristics in view of re restoration possibilities. As a conclusion some guide-lines for the use of post and core restorations in the general dental practice are formulated. PMID- 11913345 TI - [The dentist and hypertension]. AB - In order to determine the risks involved in dental treatment the blood pressure values using individual risk scores (ASA class I-IV) should be established in every patient and patients treated accordingly. In ASA class III and IV (blood pressure value 160/95 mmHg) only regular check ups and minor dental treatment is permitted; in case of major dental treatment a physician should first be consulted. ASA class IV (> or = 200/115) represents a contraindication for elective dental treatment. PMID- 11913346 TI - [Quality of dental care. The assessment of restorative treatment]. AB - An important aspect of the policy regarding quality of dental care is the possibility of assessing dental treatment. This assessment takes place in four steps: the selection of a subject, the development of criteria and standards, the collection of data and the decision whether quality is adequate. When quality is decided inadequate a plan should be developed to correct for deficiencies. After implementation quality can be reassessed. PMID- 11913347 TI - [Dentistry in Turkey and Morocco]. AB - Some information is given on the number of dentists in Turkey and Morocco and the dental condition of the inhabitants. This information concerns prevention, caries, periodontic diseases as well as other dental diseases and diseases of the mouth. PMID- 11913348 TI - [Dietary habits of Turkish and Moroccan children in the Netherlands]. AB - Migration might have considerable implications for nutrition and consequently for health. Because of the higher demands of growth, children are especially vulnerable to changes in nutrition. In order to get an insight into the need for a special nutrition policy for Turkish ad Moroccan children, the dietary habits and food consumption of 8-year-old Turkish and Moroccan children in The Netherlands were assessed. The results showed that the diet of these children was more compatible with the Dutch recommendations for a prudent diet than the diet of Dutch children from a similar socio-economic background. Quite a few of the basic food items Turkish and Moroccan children are accustomed to eat, like rice, cereals, bread, pulses and vegetables, contain valuable nutrients. So, an increased consumption of these items was recommended. The intake of calcium and riboflavin of Turkish and Moroccan children was low and so the use of dairy products--preferably in fermented form like yoghurt--was recommended. This would also enhance the Vitamin D status. PMID- 11913349 TI - [The fixed full bridge on Branemark-implants. Current developments and procedures in clinical application]. AB - In the past research in Implantology has been focused on osseointegration. Recently suprastructures get more attention. Users of the Branemark-system discuss the preference for a fixed full bridge or an overdenture. The advantages and disadvantages are enumerated in this article. The current procedures are discussed and the importance of a wax set-up is emphasized. During the impression procedure the copings are not attached to each other with acrylic. The gold cylinders, on which the suprastructure is attached to the abutments, are not soldered in the metal framework, but connected with acrylic. With the described method better results were obtained. PMID- 11913350 TI - [Saliva and saliva substitutes]. AB - Saliva is a mixture of secretions of both the large salivary glands (glandula sublingualis submandibularis and parotis) and the minor salivary glands of the palate, tongue, lips and cheeks. The rheological flow of the separate glandular salivas differs remarkably. Sublingual saliva is both more viscous and elastic than the other glandular salivas. The viscosity of parotid saliva, which is of importance during eating (digestion), is equal to the viscosity of water. Utilization of the combination of viscosity and elasticity may improve new saliva substitutes and of frequent applications of saliva substitutes by xerostomia patients may reduce. Besides, the utilization of correct visco-elastic properties of saliva substitutes may lead to better moistening and protective properties. PMID- 11913351 TI - [Quality assurance in dentistry]. AB - An important subject regarding the quality of dental care is the development of a system of quality assurance. This system should be focused on preservation and improvement of the quality of dentistry. Activities like peer review, continuing dental education, (re-)registration and professional specialization could make a major contribution to the establishment of such a system. PMID- 11913352 TI - [The promotion of the dental health of migrants]. AB - The dental health of migrants in Holland is critical. Health education can be a good method to improve this situation. It is important to correspond with the personal background, the knowledge attitude and behavior of the individual client. In health education for migrants language and cultural differences need to be overcome. Until now there is a limited number of initiatives to promote the dental health. Expert knowledge and skills of health educators need to be improved and more supporting educational materials have to be developed. PMID- 11913353 TI - [Dental treatment of migrants. Some personal experiences and advice]. AB - The treatment of migrants sometimes poses the dentist with unexpected problems. Some of these difficulties are discussed. Practical suggestions are given which may help to meet them. PMID- 11913354 TI - Who needs breasts, anyway? PMID- 11913355 TI - The lessons of railway spine. PMID- 11913356 TI - A new strategy to support the "chronic injury" model of whiplash: ignore Lithuania. PMID- 11913357 TI - High dose ratio (HDR) cervical ring applicator to control bleeding from cervical carcinoma. AB - The objective of this study was to evaluate the use of the high dose rate (HDR) cervical ring applicator to control acute cervical bleeding from carcinoma of the uterine cervix. This study consists of 15 patients presenting with invasive carcinoma of the uterine cervix with acute vaginal bleeding requiring transfusion. Initial irradiation, delivered emergently because of vaginal bleeding, consisted of two fractions (5 Gy each fraction to the surface of the cervix) utilizing the HDR intracavitary vaginal ring applicator. Two fractions were administered at one week intervals for a total of 10 Gy to the surface of the cervix. Irradiation doses from the HDR ring applications were not considered into the composite total dose to point A. Diagnostic imaging evaluation and initiation of external irradiation were commenced during this initial weekly interval between fractions. Vaginal bleeding requiring no additional transfusions was achieved in 93% (14/15) of patients. No acute or long-term Grades 3, 4, or 5 bowel or bladder sequelae were noted. In conclusion, HDR cervical ring brachytherapy is effective in controlling acute vaginal bleeding and can be delivered without undue acute or long-term toxicity. PMID- 11913359 TI - Wait watcher. Between spin and cynicism. PMID- 11913360 TI - Managers & medicine. Here's hoping.... PMID- 11913358 TI - Significance of morphometric, DNA cytometric features, and other prognostic markers on survival of endometrial cancer patients in northern Norway. AB - The objective of this study was to evaluate the prognostic value of nuclear morphometric features and DNA ploidy by flow cytometry next to depth of myometrial invasion and vascular invasion in endometrial cancer of all FIGO stages. A total of 123 women (103 FIGO stage I, eight stage II, and 12 stage III and IV) from northern Norway were studied. The follow-up period was between 7 and 19 years. The median age of patients was 62 years. The primary surgery was performed in the University Hospital of Tromso or in the seven different reference hospitals in the northern part of Norway after an endometrial cancer diagnosis. The histologic, morphometric,flow cytometric and immunohistochemical investigations were based on archival paraffin-embedded material. The information regarding the follow-up data and clinical information were obtained from the medical records. Thirteen (10.6%) patients from the entire group (all stages) but only three (2.7%) of the FIGO stage I and II patients died from locally recurrent or metastatic disease. FIGO substage (P = 0.0006; odds ratio [OR] =16.44, 95% confidence interval [CI] = 3.36-80.45), vascular invasion (P =0.01, OR = 6.42, CI = 1.57-26.34) and nuclear size (P = 0.025, OR = 1.3,CI = 1.05-1.61) were independently correlated with recurrence in a multivariate analysis but histologic grade and DNA ploidy were not. Vascular invasion was poorly reproducible both between and within the same observer, however. In this retrospective study of all stages of endometrial carcinoma with long follow-up periods the primary tumor characteristics nuclear perimeter and FIGO stage were of prognostic significance in addition to the poorly reproducible vessel invasion. PMID- 11913361 TI - Managers & medicine. Type casting. PMID- 11913362 TI - Managers & medicine. Weight of evidence. PMID- 11913364 TI - The fetal origins hypothesis. PMID- 11913363 TI - Fetal programming and the Leningrad Siege study. AB - The Leningrad Siege Study investigated the relationship between decreased maternal food intake and risk factors for coronary heart disease in adult life. The study screened 169 subjects exposed to intrauterine starvation during the Siege of Leningrad (now St. Petersburg) 1941-4, 192 subjects born in Leningrad before the siege and 188 subjects born concurrently with these two groups but outside the area of the siege. No difference was found between the subjects exposed to starvation in utero and during infancy in glucose tolerance [in utero: 5.2 mmol/l (95% confidence interval 5.1 to 5.3; infancy: 5.3 (5.1 to 5.5), p = 0.94], insulin concentration, blood pressure, lipid concentration or coagulation factors. The intrauterine exposed group had evidence of endothelial dysfunction by higher concentrations of von Willebrand factor and a stronger interaction between adult obesity and blood pressure. Non-systematic differences in subscapular to triceps skinfold ratio, diastolic blood pressure and clotting factors were demonstrated compared to the non-exposed groups. In conclusion, this study did not find an association between intrauterine starvation and glucose intolerance, dyslipidaemia, hypertension or cardiovascular disease in adult life. These findings differ from studies of subjects exposed to maternal starvation during the Dutch Hunger Winter. However, the dissimilar effects of exposure to the two famines may contribute to our understanding of the mechanisms of the thrifty phenotype and support the importance of catch-up growth during early childhood, a situation that occurred in the Netherlands by not in Leningrad. PMID- 11913365 TI - Legislative committee report to HDA members. PMID- 11913366 TI - HDA Committee profile. New dentist committee. PMID- 11913367 TI - National ADA campaign designed to reach benefit purchasers. PMID- 11913368 TI - 1966 Membership Directory. Listings for Oahu, Hawaii Kauai and Maui Members. PMID- 11913369 TI - "It seems to me...." What does the ADA do for me? PMID- 11913370 TI - County report: Kauai Dental Society. PMID- 11913371 TI - ADA position on laser for hard tissue procedures. American Dental Association Council on Scientific Affairs. PMID- 11913372 TI - Guidelines from the Working Group. Recommendations for performing transesophageal echocardiography. PMID- 11913373 TI - Incidence of pseudomembranous colitis after vancomycin-treated MRSA infection. PMID- 11913374 TI - Free-energy component analysis of 40 protein-DNA complexes: a consensus view on the thermodynamics of binding at the molecular level. AB - Noncovalent association of proteins to specific target sites on DNA--a process central to gene expression and regulation--has thus far proven to be idiosyncratic and elusive to generalizations on the nature of the driving forces. The spate of structural information on protein--DNA complexes sets the stage for theoretical investigations on the molecular thermodynamics of binding aimed at identifying forces responsible for specific macromolecular recognition. Computation of absolute binding free energies for systems of this complexity transiting from structural information is a stupendous task. Adopting some recent progresses in treating atomic level interactions in proteins and nucleic acids including solvent and salt effects, we have put together an energy component methodology cast in a phenomenological mode and amenable to systematic improvements and developed a computational first atlas of the free energy contributors to binding in approximately 40 protein-DNA complexes representing a variety of structural motifs and functions. Illustrating vividly the compensatory nature of the free energy components contributing to the energetics of recognition for attaining optimal binding, our results highlight unambiguously the roles played by packing, electrostatics including hydrogen bonds, ion and water release (cavitation) in protein-DNA binding. Cavitation and van der Waals contributions without exception favor complexation. The electrostatics is marginally unfavorable in a consensus view. Basic residues on the protein contribute favorably to binding despite the desolvation expense. The electrostatics arising from the acidic and neutral residues proves unfavorable to binding. An enveloping mode of binding to short stretches of DNA makes for a strong unfavorable net electrostatics but a highly favorable van der Waals and cavitation contribution. Thus, noncovalent protein-DNA association is a system specific fine balancing act of these diverse competing forces. With the advances in computational methods as applied to macromolecular recognition, the challenge now seems to be to correlate the differential (initial vs. final) energetics to substituent effects in drug design and to move from affinity to specificity. PMID- 11913375 TI - Theoretical calculation of the coiled-coil stability in water in the context of its possible use as a molecular rack. AB - The coiled-coil stability and rigidity may be of importance for molecular electronics (electronically bistable molecules). The coiled-coil binding free energy has been calculated using molecular dynamics (MD). The energy has been computed as a difference of the appropriate free energies; derived for the coiled coil and isolated alpha-helices separately. All MD simulations have been performed using an explicit model of the solvent, whereas the continuum solvent approach has been applied to analyze the MD trajectories. The computed stability of the coiled-coil is of the order of -87 kcal/mol, i.e., about -1.2 kcal/mol per amino acid residue, and arises mainly from the electrostatic interactions and hydrophobic effect. The entropy term has been roughly estimated to be of the order of -22 kcal/mol. This assures that coiled-coil polypeptides may be used as a stable molecular scaffolding. PMID- 11913376 TI - Maximum feasibility guideline in the design and analysis of protein folding potentials. AB - Protein folding potentials are expected to have the lowest energy for the native shape. The Linear Programming (LP) approach achieves exactly that goal for a training set, or indicates that this goal is impossible to obtain. If a solution cannot be found (i.e., the problem is infeasible) two possible routes are possible: (a) choosing a new functional form for the potential, (b) finding the best potential with a feasible subset of the data, and (or) detecting inconsistent subset of the data in the training set. Here, we explore option (b). A simple heuristic for finding an approximate solution to an infeasible set of linear inequalities is outlined. An approximately feasible solution is obtained iteratively, starting from a certain initial guess, by computing a series of analytic centers of the polyhedra defined by all the inequalities satisfied at the subsequent iterations. Standard interior point algorithms for Linear Programming can be used to compute efficiently the analytic center of a polyhedron. We demonstrate how this procedure can be used for the design of folding potentials that are linear in their parameters. The procedure shows an improvement in the quality of the potentials and sometimes points to flaws in the original data. PMID- 11913377 TI - Dynamics of large proteins through hierarchical levels of coarse-grained structures. AB - Elastic network models have been successful in elucidating the largest scale collective motions of proteins. These models are based on a set of highly coupled springs, where only the close neighboring amino acids interact, without any residue specificity. Our objective here is to determine whether the equivalent cooperative motions can be obtained upon further coarse-graining of the protein structure along the backbone. The influenza virus hemagglutinin A (HA), composed of N = 1509 residues, is utilized for this analysis. Elastic network model calculations are performed for coarse-grained HA structures containing only N/2, N/10, N/20, and N/40 residues along the backbone. High correlations (>0.95) between residue fluctuations are obtained for the first dominant (slowest) mode of motion between the original model and the coarse-grained models. In the case of coarse-graining by a factor of 1/40, the slowest mode shape for HA is reconstructed for all residues by successively selecting different subsets of residues, shifting one residue at a time. The correlation for this reconstructed first mode shape with the original all-residue case is 0.73, while the computational time is reduced by about three orders of magnitude. The reduction in computational time will be much more significant for larger targeted structures. Thus, the dominant motions of protein structures are robust enough to be captured at extremely high levels of coarse-graining. And more importantly, the dynamics of extremely large complexes are now accessible with this new methodology. PMID- 11913378 TI - Rapid grid-based construction of the molecular surface and the use of induced surface charge to calculate reaction field energies: applications to the molecular systems and geometric objects. AB - This article describes a number of algorithms that are designed to improve both the efficiency and accuracy of finite difference solutions to the Poisson Boltzmann equation (the FDPB method) and to extend its range of application. The algorithms are incorporated in the DelPhi program. The first algorithm involves an efficient and accurate semianalytical method to map the molecular surface of a molecule onto a three-dimensional lattice. This method constitutes a significant improvement over existing methods in terms of its combination of speed and accuracy. The DelPhi program has also been expanded to allow the definition of geometrical objects such as spheres, cylinders, cones, and parallelepipeds, which can be used to describe a system that may also include a standard atomic level depiction of molecules. Each object can have a different dielectric constant and a different surface or volume charge distribution. The improved definition of the surface leads to increased precision in the numerical solutions of the PB equation that are obtained. A further improvement in the precision of solvation energy calculations is obtained from a procedure that calculates induced surface charges from the FDPB solutions and then uses these charges in the calculation of reaction field energies. The program allows for finite difference grids of large dimension; currently a maximum of 571(3) can be used on molecules containing several thousand atoms and charges. As described elsewhere, DelPhi can also treat mixed salt systems containing mono- and divalent ions and provide electrostatic free energies as defined by the nonlinear PB equation. PMID- 11913379 TI - Folding funnels: the key to robust protein structure prediction. AB - Natural proteins fold because their free energy landscapes are funneled to their native states. The degree to which a model energy function for protein structure prediction can avoid the multiple minima problem and reliably yield at least low resolution predictions is also dependent on the topography of the energy landscape. We show that the degree of funneling can be quantitatively expressed in terms of a few averaged properties of the landscape. This allows us to optimize simplified energy functions for protein structure prediction even in the absence of homology information. Here we outline the optimization procedure in the context of associative memory energy functions originally introduced for tertiary structure recognition and demonstrate that even partially funneled landscapes lead to qualitatively correct, low-resolution predictions. PMID- 11913380 TI - Identifying native-like protein structures using physics-based potentials. AB - As the field of structural genomics matures, new methods will be required that can accurately and rapidly distinguish reliable structure predictions from those that are more dubious. We present a method based on the CHARMM gas phase implicit hydrogen force field in conjunction with a generalized Born implicit solvation term that allows one to make such discrimination. We begin by analyzing pairs of threaded structures from the EMBL database, and find that it is possible to identify the misfolded structures with over 90% accuracy. Further, we find that misfolded states are generally favored by the solvation term due to the mispairing of favorable intramolecular ionic contacts. We also examine 29 sets of 29 misfolded globin sequences from Levitt's "Decoys 'R' Us" database generated using a sequence homology-based method. Again, we find that discrimination is possible with approximately 90% accuracy. Also, even in these less distorted structures, mispairing of ionic contacts results in a more favorable solvation energy for misfolded states. This is also found to be the case for collapsed, partially folded conformations of CspA and protein G taken from folding free energy calculations. We also find that the inclusion of the generalized Born solvation term, in postprocess energy evaluation, improves the correlation between structural similarity and energy in the globin database. This significantly improves the reliability of the hypothesis that more energetically favorable structures are also more similar to the native conformation. Additionally, we examine seven extensive collections of misfolded structures created by Park and Levitt using a four-state reduced model also contained in the "Decoys 'R' Us" database. Results from these large databases confirm those obtained in the EMBL and misfolded globin databases concerning predictive accuracy, the energetic advantage of misfolded proteins regarding the solvation component, and the improved correlation between energy and structural similarity due to implicit solvation. Z-scores computed for these databases are improved by including the generalized Born implicit solvation term, and are found to be comparable to trained and knowledge-based scoring functions. Finally, we briefly explore the dynamic behavior of a misfolded protein relative to properly folded conformations. We demonstrate that the misfolded conformation diverges quickly from its initial structure while the properly folded states remain stable. Proteins in this study are shown to be more stable than their misfolded counterparts and readily identified based on energetic as well as dynamic criteria. In summary, we demonstrate the utility of physics-based force fields in identifying native-like conformations in a variety of preconstructed structural databases. The details of this discrimination are shown to be dependent on the construction of the structural database. PMID- 11913381 TI - Computational alanine scanning of the 1:1 human growth hormone-receptor complex. AB - The MM-PBSA (Molecular Mechanics-Poisson-Boltzmann surface area) method was applied to the human Growth Hormone (hGH) complexed with its receptor to assess both the validity and the limitations of the computational alanine scanning approach. A 400-ps dynamical trajectory of the fully solvated complex was simulated at 300 K in a 101 A x 81 A x 107 A water box using periodic boundary conditions. Long-range electrostatic interactions were treated with the particle mesh Ewald (PME) summation method. Equally spaced snapshots along the trajectory were chosen to compute the binding free energy using a continuum solvation model to calculate the electrostatic desolvation free energy and a solvent-accessible surface area approach to treat the nonpolar solvation free energy. Computational alanine scanning was performed on the same set of snapshots by mutating the residues in the structural epitope of the hormone and the receptor to alanine and recomputing the deltaGbinding. To further investigate a particular structure, a 200-ps dynamical trajectory of an R43A hormone-receptor complex was simulated. By postprocessing a single trajectory of the wild-type complex, the average unsigned error of our calculated deltadeltaGbinding is approximately1 kcal/mol for the alanine mutations of hydrophobic residues and polar/charged residues without buried salt bridges. When residues involved in buried salt bridges are mutated to alanine, it is demonstrated that a separate trajectory of the alanine mutant complex can lead to reasonable agreement with experimental results. Our approach can be extended to rapid screening of a variety of possible modifications to binding sites. PMID- 11913382 TI - Is there a unique melting temperature for two-state proteins? AB - Thermal unfolding (or folding) in many proteins occurs in an apparent two-state manner, suggesting that only two states, unfolded and folded, are populated. At the melting temperature, Tm, the two states coexist. Using lattice models with side chains we show that individual residues become structured at temperatures that deviate from Tm, which implies that partially folded conformations make substantial contribution to thermodynamic properties of two-state proteins. We also find that the folding cooperativity for a given residue is linked to its accessible surface area. These results are consistent with the experiments on GCN4-like zipper peptide, which showed that local melting temperatures differ from Tm. Analysis of thermal unfolding of six proteins shows that deltaT/Tm approximately N(-1), where deltaT is the transition width and N is the number of residues. This scaling allows us to conclude that, when corrected for finite size effects, folding cooperativity can be captured using coarse grained models. PMID- 11913383 TI - Polymer collapse, protein folding, and the percolation threshold. AB - We study the transition of polymers in the dilute regime from a swollen shape at high temperatures to their low-temperature structures. The polymers are modeled by a single self-avoiding walk (SAW) on a lattice for which l of the monomers (the H monomers) are self-attracting, i.e., if two nonbonded H monomers become nearest neighbors on the lattice they gain energy of interaction (epsilon = /epsilon/); the second type of monomers, denoted P, are neutral. This HP model was suggested by Lau and Dill (Macromolecules 1989, 22, 3986-3997) to study protein folding, where H and P are the hydrophobic and polar amino acid residues, respectively. The model is simulated on the square and simple cubic (SC) lattices using the scanning method. We show that the ground state and the sharpness of the transition depend on the lattice, the fraction g of the H monomers, as well as on their arrangement along the chain. In particular, if the H monomers are distributed at random and g is larger than the site percolation threshold of the lattice, a collapsed transition is very likely to occur. This conclusion, drawn for the lattice models, is also applicable to proteins where an effective lattice with coordination number between that of the SC lattice and the body centered cubic lattice is defined. Thus, the average fraction of hydrophobic amino acid residues in globular proteins is found to be close to the percolation threshold of the effective lattice. PMID- 11913384 TI - Computation of the physio-chemical properties and data mining of large molecular collections. AB - Very large data sets of molecules screened against a broad range of targets have become available due to the advent of combinatorial chemistry. This information has led to the realization that ADME (absorption, distribution, metabolism, and excretion) and toxicity issues are important to consider prior to library synthesis. Furthermore, these large data sets provide a unique and important source of information regarding what types of molecular shapes may interact with specific receptor or target classes. Thus, the requirement for rapid and accurate data mining tools became paramount. To address these issues Pharmacopeia, Inc. formed a computational research group, The Center for Informatics and Drug Discovery (CIDD).* In this review we cover the work done by this group to address both in silico ADME modeling and data mining issues faced by Pharmacopeia because of the availability of a large and diverse collection (over 6 million discrete compounds) of drug-like molecules. In particular, in the data mining arena we discuss rapid docking tools and how we employ them, and we describe a novel data mining tool based on a ID representation of a molecule followed by a molecular sequence alignment step. For the ADME area we discuss the development and application of absorption, blood-brain barrier (BBB) and solubility models. Finally, we summarize the impact the tools and approaches might have on the drug discovery process. PMID- 11913385 TI - Qmd-plot: a graphical utility for rapid preliminary analysis of time series of fluctuating data, developed in the context of molecular dynamics simulations. AB - Qmd-plot is a utility to obtain rapid information about past or on-going simulations, or real-time data collections, in the form of graphs of recorded variables (x, y, ...), as x-y plots or as a function of simulated or real time. Time series records in the data file must be named. Variable names and their locations in the data file are initially unknown to the program, but are identified in a first scan, in which header records are located on the basis of a predefined key (that can be changed interactively). The names of the time series are then presented in an interactive menu, from which the user can repeatedly specify a graph to be viewed. Qmd-plot has been developed in the context of molecular dynamics simulations. We give examples of time series and x-y plots made from output of the sigma program. Qmd-plot code is a Java application; source and class files can be obtained free from the authors. PMID- 11913386 TI - Docking of small ligands to low-resolution and theoretically predicted receptor structures. AB - We have developed a simple docking procedure that is able to utilize low resolution models of proteins created by structure prediction algorithms such as threading or ab initio folding to predict the conformation of receptor-small ligand complexes. In our approach, using only approximate, discretized models of both molecules, we search for the steric and quasi-chemical complementarity between a ligand and the receptor molecules. This averaging procedure allows for the compensation of numerous structural inaccuracies resulting from the theoretical predictions of the receptor structure. The best relative orientation of these two models is obtained by an exhaustive scan over the rigid body's six dimensional translational and rotational degrees of freedom. The search method is based on a real space grid-searching algorithm, unlike docking methods based on the fast Fourier Transform algorithm. We have applied this algorithm to rebuild structures of several complexes available in the Protein Data Bank. The structures of the receptors are produced by means of our threading algorithm PROSPECTOR, subsequently refined, and then utilized in the docking experiment. In many cases, not only is the localization of the binding site on the receptor surface correctly identified, but the proper orientation of the bounded ligand is also reasonably well reproduced within the level of accuracy of the modeled receptor itself. PMID- 11913387 TI - Evolution of physics-based methodology for exploring the conformational energy landscape of proteins. AB - The evolution of our physics-based computational methods for determining protein conformation without the introduction of secondary-structure predictions, homology modeling, threading, or fragment coupling is described. Initial use of a hard-sphere potential captured much of the structural properties of polypeptide chains, and subsequent more refined force fields, together with efficient methods of global optimization provide indications that progress is being made toward an understanding of the interresidue interactions that underlie protein folding. PMID- 11913388 TI - Predicted solution structure of zymogen human coagulation FVII. AB - A model solution structure for the complete tissue factor-free calcium ion-bound human zymogen FVII (residues 1-406) (FVII) has been constructed to study possible conformational changes associated with the activation process and tissue factor (TF) binding. The initial structure for the present model was constructed using the X-ray crystallographic structure of human coagulation FVIIa/TF complex bound with calcium ions (Banner et al., Nature 1996, 380, 41-46). This model was subsequently subjected to lengthy molecular dynamics simulations. The Amber force field in conjunction with the PME electrostatic summation method was employed. The estimated TF free solution structure was then compared with the currently available X-ray crystal structures of FVIIa (with or without TF, variable inhibitor bound) to estimate the restructuring of FVII due to TF binding and activation. The solution structure of the zymogen FVII in the absence of TF is predicted to be an extended domain structure similar to that of the TF-bound X ray crystal structure. An additional extension of the serine protease (SP) domain of the zymogen above a reference lipid surface by approximately 7 A was in agreement with experiment. Significant Gla-EGF1 and EGF1-EGF2 interdomain motions in the zymogen were observed. Carbohydrate dimers attached to Ser-52 and Ser-60 did not cause restructuring in this domain. Minimal restructuring of the SP domain is found upon inference of the zymogen from the activated form. The catalytic triad residues maintain the H-bonded network while Lys-341 occupies the S1 specific site in the zymogen. PMID- 11913389 TI - Simulating enzyme reactions: challenges and perspectives. AB - Elucidating how enzymes enhance the rates of the reactions that they catalyze is a major goal of contemporary biochemistry, and it is an area in which computational and theoretical techniques can make a major contribution. This article outlines some of the processes that need to be investigated if enzyme catalysis is to be understood, reviews the current state-of-the-art in enzyme simulation work, and highlights challenges for the future. PMID- 11913390 TI - Computational studies of reaction mechanisms of methane monooxygenase and ribonucleotide reductase. AB - An overview of the computational efforts made by our group during the last few years in the field of nonheme diiron proteins is presented. Through application of ab initio methodology to a reasonable set of molecular models, significant progress is made in understanding how the soluble Methane Monooxygenase system achieves the hydroxylation of methane and how the catalytic cycle of Ribonucleotide Reductase is initiated. In particular, the current studies reveal in more detail (1) the nature of key intermediates in the reaction cycles of these two metalloenzymes, (2) details of how the iron centers regulate the systems, and (3) important aspects of how the carboxylate ligands in the active sites may tailor the enzymatic needs of the metalloprotein. This knowledge also leads to novel connections between the two enzymes. The coordinative unsaturation and carboxylate shifts investigated herein are two properties that are likely to be of more general impact in nonheme proteins. The control of the redox chemistry of the enzyme by the binuclear metal center, also analyzed here, should find common ground among other bimetallic systems as well. PMID- 11913391 TI - The elastic net algorithm and protein structure prediction. AB - Predicting protein structures from their amino acid sequences is a problem of global optimization. Global optima (native structures) are often sought using stochastic sampling methods such as Monte Carlo or molecular dynamics, but these methods are slow. In contrast, there are fast deterministic methods that find near-optimal solutions of well-known global optimization problems such as the traveling salesman problem (TSP). But fast TSP strategies have yet to be applied to protein folding, because of fundamental differences in the two types of problems. Here, we show how protein folding can be framed in terms of the TSP, to which we apply a variation of the Durbin-Willshaw elastic net optimization strategy. We illustrate using a simple model of proteins with database-derived statistical potentials and predicted secondary structure restraints. This optimization strategy can be applied to many different models and potential functions, and can readily incorporate experimental restraint information. It is also fast; with the simple model used here, the method finds structures that are within 5-6 A all-Calpha-atom RMSD of the known native structures for 40-mers in about 8 s on a PC; 100-mers take about 20 s. The computer time tau scales as tau approximately n, where n is the number of amino acids. This method may prove to be useful for structure refinement and prediction. PMID- 11913392 TI - Transition state docking: a probe for noncovalent catalysis in biological systems. Application to antibody-catalyzed ester hydrolysis. AB - A strategy for pinpointing favorable noncovalent interactions between transition states and active sites of biological catalysts is described. This strategy combines high-level quantum mechanical calculations of transition state geometries with an automated docking procedure using AutoDock. By applying this methodology to antibody-catalyzed hydrolyses of aryl esters (by the 48G7, CNJ206, and 17E8 families of antibodies), varying levels of catalysis are explained in terms of specific hydrogen bonding interactions between combining site residues and transition states. Although these families of antibodies were produced in separate experiments by different researchers using related but different haptens, the mechanism of transition state stabilization appears to be highly conserved. Despite being elicited in response to anionic phosphonate haptens, the best catalysts often utilize hydrogen bond acceptors to stabilize transition states. A mutant of antibody CNJ206, designed based on this observation and predicted to be a better catalyst, is proposed. In the case of antibody 48G7, affinity maturation is shown to produce a catalyst that is highly selective for one of two enantiomeric transition states from a nonselective germline precursor. PMID- 11913393 TI - Combined QM/MM study of the opsin shift in bacteriorhodopsin. AB - Combined quantum mechanical and molecular mechanical (QM/MM) calculations and molecular dynamics simulations of bacteriorhodopsin (bR) in the membrane matrix have been carried out to determine the factors that make significant contributions to the opsin shift. We found that both solvation and interactions with the protein significantly shifts the absorption maximum of the retinal protonated Schiff base, but the effects are much more pronounced in polar solvents such as methanol, acetonitrile, and water than in the protein environment. The differential solvatochromic shifts of PSB in methanol and in bR leads to a bathochromic shift of about 1800 cm(-1). Because the combined QM/MM configuration interaction calculation is essentially a point charge model, this contribution is attributed to the extended point-charge model of Honig and Nakanishi. The incorporation of retinal in bR is accompanied by a change in retinal conformation from the 6-s-cis form in solution to the 6-s-trans configuration in bR. The extension of the pi-conjugated system further increases the red-shift by 2400 cm(-1). The remaining factors are due to the change in dispersion interactions. Using an estimate of about 1000 cm(-1) in the dispersion contribution by Houjou et al., we obtained a theoretical opsin shift of 5200 cm( 1) in bR, which is in excellent agreement with the experimental value of 5100 cm( 1). Structural analysis of the PSB binding site revealed the specific interactions that make contributions to the observed opsin shift. The combined QM/MM method used in the present study provides an opportunity to accurately model the photoisomerization and proton transfer reactions in bR. PMID- 11913394 TI - Medication error prevention--potassium chloride. PMID- 11913395 TI - Accreditation Committee approves examples of voluntarily reportable sentinel events. PMID- 11913396 TI - A look back at Congressional action 2001. PMID- 11913398 TI - Transactions delay prompts concern among covered entities. PMID- 11913397 TI - The transactions extension: no reason to procrastinate. PMID- 11913399 TI - Help wanted: five steps to take before you hire. PMID- 11913400 TI - Practice brief. Retaining healthcare business records. American Health Information Management Association. PMID- 11913401 TI - Web-based system improves access. PMID- 11913402 TI - Merghers and acquisitions: they could happen to you. PMID- 11913403 TI - Implementing HIPAA one step at a time. PMID- 11913404 TI - Putting together the privacy puzzle. Interview by Jane Jeffries. PMID- 11913405 TI - Pamela Loving: creating opportunity for others. Interview by Laurie Larson. PMID- 11913406 TI - Points to consider for the development of new indications for hormone replacement therapies and estrogen-like molecules. Department of Urogynaecology, King's College Hospital, London. PMID- 11913407 TI - Effects of different hormone replacement regimens on postmenopausal women with abnormal lipid levels. Menopause Study Group. AB - OBJECTIVES: A retrospective, subset analysis of the prospective Menopause Study Group data was performed to determine the effects of four conjugated estrogens (CE) + medroxyprogesterone acetate (MPA) regimens and one unopposed CE regimen on the lipid profiles of women with abnormal lipid levels (n = 525). Previously unpublished data on the entire study population were also reviewed to determine the effects of these hormone replacement regimens on blood pressure and weight (n = 1368, 1374, respectively). METHODS: During a 1-year, prospective trial, all patients took CE. Groups A and B also took continuous MPA 2.5 and 5 mg, respectively, C and D took MPA 5 and 10 mg for the last 14 cycle days of each 28 day cycle, and E took matching placebo tablets to replace MPA. RESULTS: After 1 year, all five regimens were directly associated with significant elevations in high density lipoprotein-2 (HDL2) cholesterol and significant reductions in low density lipoprotein (LDL) cholesterol (p < 0.05), although unopposed estrogen produced the greatest increase in HDL2 cholesterol. The CE + MPA regimens were also associated with significant decreases in total cholesterol. Triglyceride levels did not change significantly from baseline in any treatment group (baseline values > 1.808 mmol/l (> 160 mg/dl)), although the CE + MPA regimens were associated with a mean triglyceride decrease and CE-only women (n = 8) had a mean triglyceride increase. No regimen produced significant weight or blood pressure changes. CONCLUSIONS: This analysis suggests that hormone replacement therapy (CE + MPA), used in appropriate dosages, may alter the HDL2 cholesterol and LDL cholesterol lipoproteins in postmenopausal women with borderline lipid levels (a higher-risk population) in a cardioprotective manner. PMID- 11913408 TI - Amenorrhea frequency with continuous combined hormone replacement therapy: a retrospective analysis. Menopause Study Group. AB - OBJECTIVES: The study retrospectively determined in postmenopausal women the cumulative amenorrhea patterns with continuous conjugated equine estrogens (CE) with or without continuous medroxyprogesterone acetate (MPA), and withdrawal bleeding patterns with cyclic therapy. METHODS: During a 1-year, prospective trial, all patients took CE. Groups A and B also took continuous MPA 2.5 and 5 mg, respectively, C and D took MPA 5 and 10 mg for the last 14 cycle days, respectively, and E took placebo to match MPA. RESULTS: By cycle 7, 40.4, 52.6 and 53.8% of women in Groups A, B and E, respectively, became amenorrheic. Percentages increased at each subsequent cycle. Of the remaining women in Groups A and B, 42.1 and 46.2%, respectively, experienced only spotting at cycle 7. In Groups C and D, 80 and 65.9%, respectively, had a mean change of equal to or less than 3 days in the onset of withdrawal bleeding/spotting from cycle to cycle for all 13 evaluable cycles. CONCLUSIONS: Amenorrhea is often desirable, and a continuous combined daily regimen of CE 0.625 mg + MPA 2.5 or 5 mg, which resulted in progressive increases in amenorrheic frequency during the 1-year treatment period, may promote compliance. In some women, however, the cyclic regimens, which produced fairly predictable withdrawal bleeding, may be preferable. PMID- 11913409 TI - Menopausal symptom control and side-effects on continuous estrone sulfate and three doses of medroxyprogesterone acetate. Ogen/Provera Study Group. AB - OBJECTIVES: To establish the optimum oral daily dose of micronized medroxyprogesterone acetate (MPA), given in combination with 1.25 mg of estrone sulfate for menopausal symptom control. METHODS: This multicenter, randomized, double-blind study was conducted on 568 postmenopausal women who were randomized to take estrone sulfate 1.25 mg daily with 2.5, 5.0 or 10 mg of MPA daily for 2 years. The number of vasomotor symptoms and the severity of mood swings, lethargy, vaginal dryness and loss of libido as well as side-effects were recorded in a diary. Blood pressure and weight were recorded at each 3-month visit. RESULTS: Vasomotor symptoms were reported by approximately 80% of subjects at month 1, 23% at month 3 but only 9% by month 24. Mood swings, lethargy and vaginal dryness improved rapidly in the initial 3 months of therapy. Decrease in libido had a slower response to therapy in all three treatment groups. Breast tenderness was the commonest side-effect with 22% of subjects complaining of this in the first 3 months of therapy, dropping to 13% by 6 months. Headache, depression, nausea, bloating and irritability showed a similar pattern of decline. There was no significant difference in the rate of decrease in menopausal symptoms or reported side-effects between the three treatment groups. There was a small but significant (p < 0.001) decrease in systolic and diastolic blood pressure over the study period. CONCLUSIONS: All three treatment regimens provide adequate symptom control. Side-effects decreased markedly after the first 3 months, with no significant difference between the treatment groups. PMID- 11913411 TI - Interleukin-2-Deficient mice: effect on cytokines and inflammatory cells in chronic colonic disease. AB - Mice deficient in interleukin-2 (IL-2-/-) develop inflammatory bowel disease resembling human ulcerative colitis. After death, macroscopic and microscopic scores were used to determine colonic inflammation. Both scores were significantly increased in the colon of IL-2-/- mice as compared to wild types mice. The level of IL-1beta 24-week-old was increased in IL-2-/- mice produced by the colon as compared with IL-2+/+ controls. However, the concentrations of IL-6 and IL-10 were not changed. The spleen weight of IL-2-/- mice was significantly increased compared with IL-2+/+ controls. We used immunochemical techniques in low-temperature paraffin-embedded spleen of IL-2-/- mice to examine pathological changes of CD4+ T cells, CD8' T cells, and CD11b+ cells. The tissue was successfully stained and was well preserved. The percentage CD4+ T cells was not significantly changed, while the percentage CD8+ T cells was significantly decreased in IL-2-/- mice compared with IL-2+/+ controls. On the other hand, the percentage CD11b+ cells was significantly increased in the spleen of IL-2-/- mice compared with IL-2+/- controls. As well as the marked difference in CD8+ and CD11b+ cells in the spleen, the increased level of IL-1beta in colonic tissue might indicate that cytotoxic T cells as well as macrophages are involved in the development and/or perpetuation of the inflammatory reactions in IL-2-/- mice. PMID- 11913410 TI - Neuropeptides and nerve growth in inflammatory bowel diseases: a quantitative immunohistochemical study. AB - Many studies have indicated changes in neuropeptides in inflammatory bowel disease (IBD), but with contradictory results. Nerve growth factor also has a potential role in the maintenance of enteric nerves and may be associated with IBD. A quantitative immunohistochemical method was used to measure area density of immunoreactive nerves in the colonic mucosa of surgical specimens. No significant differences in immunoreactivity for substance P, vasoactive intestinal polypeptide, growth associated protein 43, and the neurotrophin receptor p75 were seen in the control, Crohn's, and ulcerative colitis groups. Compared to age-matched normal colon (N = 18), there was an increase in neutrophil number in Crohn's (P < 0.05) and ulcerative colitis (P < 0.01) (both N = 9). There were positive correlations (P < 0.05) between neutrophil number and growth associated protein, between p75 and substance P immunoreactive nerves in ulcerative colitis, and between p75 and vasoactive intestinal polypeptide in Crohn's specimens. These data indicate a link between the immunologic and nervous systems in IBD. PMID- 11913412 TI - Validation of Spanish language dyspepsia questionnaire. AB - No dyspepsia-specific questionnaire currently exists in Spanish. The Spanish Language Dyspepsia Questionnaire (SLDQ) was developed based on Rome dyspepsia criteria, other questionnaires, and common symptoms. Self-reported normal and dyspeptic volunteers (N = 63) in Chiapas, Mexico, participated in a validation study. We assessed intra- and interrater reliability by test-retest studies and established validity by both correlation to the Short Form-36 (SF-36) and comparison of scores between normals and dyspeptics. The total SLDQ score showed a wide distribution (range 0-78, mean 23.7 +/- 21.9). Internal reliability of the SLDQ was high (Cronbach's a = 0.93). Intra- and interrater reliability were excellent (scores from the first and second interviews not statistically different; P = 0.94; intraclass correlation coefficient = 0.96). SLDQ scales correlated appropriately with the SF-36. The SLDQ distinguished self-classified normals from dyspeptics (P < 0.001). The SLDQ fills the unmet need for a valid, reproducible, and multidimensional Spanish-language instrument to measure dyspepsia. Additionally, we have made suggestions for the development of symptom quantifying questionnaires. PMID- 11913414 TI - Improving outcomes in depression. Practice of medicine should carry health warning. PMID- 11913413 TI - Is glutamate involved in transient lower esophageal sphincter relaxations? AB - Glutamate is an important excitatory amino acid and plays a major role in brain stem neurotransmission. Although the effect of glutamate on esophageal motility is well studied, its role in the triggering of transient lower esophageal sphincter relaxations (TLESRs) remains to be determined. Esophageal manometry was performed in 10 healthy volunteers using a perfused sleeve assembly. The effect of intragastric instillation of the nonspecific N-methyl-D-aspartate receptor antagonist dextromethorphan (30 mg) and the glutamate-release inhibitor riluzole (100 mg) was evaluated on esophageal motility and on the rate of TLESRs during isovolumetric gastric distension (500 ml). Dextromethorphan and riluzole had no effect on the amplitude or peristaltic velocity of esophageal pressure waves, basal LES pressure, or LES relaxation after water swallowing. Gastric distension increased the rate of TLESRs from 2.0 (1.0-3.5)/45 min to 5.0 (4.0-7.0)/45 min during placebo (P < 0.05). In contrast, the rate of TLESRs during gastric distension was significantly reduced with riluzole [4.0 (2.5-6.0)/45 min], but not with dextromethorphan. In conclusion, riluzole had no effect on swallow induced LES relaxation, esophageal peristalsis, or gastric tone, but it reduced the number of TLESRs evoked by gastric distension. These findings suggest that glutamate may be involved in the neurocircuitry underlying TLESRs. However, as the effect was only marginal, additional studies are required to confirm our observations. PMID- 11913415 TI - The future of rehabilitation. Specialist rehabilitation is best done by those with skills in rehabilitation. PMID- 11913416 TI - Response and adaptation of beagle dogs to hypergravity. AB - Eight male Beagle dogs, five months old, were centrifuged for three months at progressively increasing loads ranging from 1.5 g to 2.5 g on a 7.9 m radius centrifuge. Dogs were maintained on the centrifuge, which was run almost continuously, by employing a special designed automated waste disposal and life support system. Heart rate and deep body temperature were monitored continuously by implant biotelemetry. Initially, centrifuged dogs showed transient decreases in heart rate and body temperature along with changes in their diurnal rhythm patterns. Compared with normal gravity controls, exposed dogs showed a slower growth rate and a reduced amount of body fat. Blood protein, total lipids, cholesterol, calcium, packed cell volume, red blood cell count, and hemoglobin were also decreased significantly. Absolute weights of the leg bones of centrifuged dogs were significantly greater than controls. Photon absorptiometry revealed significant density increases in selective regions of the femur and humerus of centrifuged dogs. In spite of the various changes noted, results from this and other studies affirm the view that dogs can tolerate and adapt to sustained loads as high as 2.5 g without serious impairment of their body structure and function. PMID- 11913417 TI - Flux of high-LET cosmic-ray particles in manned space flight. AB - On the Apollo and Skylab missions the high energy heavy ion (HZE) flux was measured by means of plastic nuclear track detectors. Measurements involve the fluxes of high linear energy transfer (LET), 6 < or approximately Z < or approximately 26 particles incident on astronauts and on several biological experiments. Partial results of these measurements are presented; the effects of shielding and solar modulation are discussed. PMID- 11913418 TI - Physical dosimetric evaluations in the Apollo 16 microbial response experiment. AB - Nine biological species, including viruses, bacteria, fungi, and nematodes, were exposed to various combinations of space vacuum, galactic radiation, and solar UV light during the Apollo 16 space flight. No major changes in number of surviving cells occurred, permitting detailed genetic and somatic studies of returned test subjects. To enable dose-response studies, solar UV was employed as a mutagenic source with cells exposed to full sunlight or to components of the UV spectrum at peak wavelengths of 254, 280, and 300 nanometers over a range of energy levels. Proper in-flight UV irradiation monitoring required the development of a potassium ferrioxalate actinometer and an anaerobic photographic emulsion dosimeter which were tested for the first time in space. Studies of the mutagenic activity of cosmic-ray particulate radiation environment required measurement of its components with several lithium fluoride thermoluminescent dosimeters and a package of passive nuclear-track detectors capable of recording high-energy multicharge particles. These detectors included cellulose nitrate, Lexan, Ilford G5, and silver chloride crystals. The nuclear track detectors measured the incident heavy particles with the recorded spherical fluences with LET350,H2O>100 keV x micrometers-1 to be 19.3 +/- 1.8 particles cm-2. This value was found to be lower than that recorded by detectors located in the Biostack and the passive personnel dosimeters worn by the astronauts, suggesting a somewhat greater average shielding. PMID- 11913419 TI - Effects of solar ultraviolet radiations on Bacillus subtilis spores and T7 bacteriophage. AB - Spores of Bacillus subtilis HA 101 and the DNA polymerase I-defective mutant HA 101 (59) F were exposed to selected wavelengths of solar ultraviolet light and space vacuum during the return of Apollo 16. In addition, coliphage T7 suspensions were exposed to solar ultraviolet radiation as part of the Microbial Response to Space Environment Experiment. Optical filters were employed to provide different energy levels at wavelengths 254 nm and 280 nm. Dose-response curves for lethal and mutagenic effects were compared with ground-based data. A close parallel was observed between the results of solar radiation and ground tests with spores of the two strains. However, significantly greater inactivation of T7 bacteriophage was observed after exposure to solar ultraviolet radiation. PMID- 11913420 TI - Radiobiological results of the Biostack experiment on board Apollo 16 and 17. AB - After penetrating the Biostack capsule, some of the HZE particles hit the biological objects carried: bacterial spores (Bacillus subtilis), seeds (Arabidopsis thaliana and Vicia faba), and shrimp eggs (Artemia salina). The different biological objects were affected by heavy ions in widely varying ways. A broad range of radiobiological investigations has been carried out in regard to the objects' response to HZE particles. The most sensitive biological objects in the Biostack experiments proved to be the shrimp eggs. The development of 500 eggs hit by heavy cosmic ions was investigated. This differed significantly from the flight controls (eggs flown in the Biostack but not hit by heavy ions) and from the ground controls. From this it has been concluded that penetration on the part of a single heavy ion may injure the encysted blastula. This damage was found to influence gastrula formation and even the hatching process of the nauplius. Abnormalities (increased by a factor of 10) in the orthonauplius were observed during the development of the hit eggs; they consisted, for example, of shortened extremities or an abnormal thorax or abdomen. In addition, eggs of Tribolium confusum and Carausius morosus, which were included in Biostack 2 (Apollo 17), have been investigated, and the influence of single heavy ions on the development process of these highly organized insects has been studied. PMID- 11913421 TI - Results of the Bacillus subtilis unit of the Biostack II experiment: physical characteristics and biological effects of individual cosmic HZE particles. AB - The effectiveness of cosmic HZE-particles on unicellular procaryotic, organisms was studied on Bacillus subtilis spores, which were accommodated in the Biostack I and II experiments on board Apollo 16 and 17. Identification of the spores that were hit was achieved by using the Biostack sandwich construction and by precise microscopical measurements of tracks of particles. Germination, outgrowth and the rate of cellular elongation were investigated. A method was developed to determine the charge of each individual HZE particle that penetrated a spore and its energy loss in the region of hit. An attempt was made to establish a connection between these physical characteristics and the biological effects produced. PMID- 11913422 TI - Modifying effect of dynamic space flight factors on radiation damage of air-dry seeds of Crepis capillaris (L) Wallr. AB - The influence of dynamic factors (vibration and linear acceleration) on the rate of chromosome aberrations in Crepis capillaris was studied. The vibrational process simulated was similar in its characteristics to that occurring at the launch of spaceships. In combination with linear acceleration it caused a statistically significant increase in the rate of chromosome aberrations compared with the control (R=7.70). The dynamic factors modified the effect of radiation damage induced by acute gamma-irradiation (3 krad). Pre-radiation treatment with vibration and acceleration on the seeds caused a significant decrease (R=10.23) of the effect of radiation damage, from 15.57% to 9.74%. The post-radiation treatment of C. capillaris seeds with the dynamic factors did not change the rate of chromosome aberrations significantly (from 15.57% to 15.90%). PMID- 11913424 TI - Gravitational effects on body composition in birds. AB - Gallinaceous birds, presenting a wide range of body size, were adapted physiologically to hyperdynamic environments, provided by chronic centrifugation. Chemical composition was measured directly on prepared carcasses, which were anatomically comparable, and more amenable to analysis than the intact body. Body mass and body fat decreased arithmetically with increasing field strength and also with increasing body mass. Water content of lean tissue increased in hyperdynamic environments, but irrespectively of body size. PMID- 11913423 TI - Effects of space balloon flights on reproductive activity in Paramecium aurelia. AB - Post autogamous Paramecium aurelia cultures were placed in hermetic containers, including a heating device with accuracy kept around +/- 0.1 degrees C. Kinetics of cellular growth was determined by cell count, after recovery, on in-flight cultures and ground control cultures. Dosimetry was performed by thermoluminescent detectors (CaSO4 activated with dysprosium). Flight durations of maximum altitude (ceiling) ranged between 48 min and 15 hours (repeated flights). Conclusions are as follows: short flights result in a secondary stimulating effect, shown by post-flight increase of the growth rate (total dose above 2 mrads); long flights or repeated flights are accompanied by a decrease in growth rate (total dose ranging from 2 to 6 mrads); in the stimulation experiments, cell counts performed immediately after flight permit identification of a temporary decrease of growth rate. The biphasic character of the biological response after flights may be due to an ionization phenomena induced by cosmic rays. Indeed, the temporary drop of growth rate is not observed after recovery if the cells are subcultured in fresh medium and left on the earth's surface. We observe, on the contrary, an increase in growth rate. These findings confirm the great sensitivity of Paramecium aurelia to very low doses of ionizing radiations and demonstrate the biological effect of cosmic radiation. PMID- 11913425 TI - The influence of variable gravitational fields on the embryonic development of some ecaudate amphibians. AB - The fertilized eggs of the frog Rana temporaria immediately after fertilization were rotated on a clinostat in the vertical plane at the rate of 0.66 rotations per minute with a radius of 20 mm for 2.5 hours. A wide spectrum of developmental anomalies was found (33% in the experiment, 14% in the control) which, in the authors' opinion, result from abnormalities in the cortical reaction of symmetrization. These abnormalities manifest themselves in the irregular distribution of cortical pigment and in eccentric division lines. PMID- 11913426 TI - Life sciences and space research XIII. Proceedings of the Open Meeting of the Working Group on Space Biology of the Seventeenth Plenary Meeting of COSPAR, Sao Paulo, S.P., Brazil--June 1974. PMID- 11913427 TI - Human sensitivity to gravity (on the problem of gravipreferendum). AB - Experiments were carried out in a centrifuge with an arm of 7.25 m, the cabin being equipped with a self-contained system of control. During an exposure to head-to-feet accelerations (+gz) of 2 g and an onset rate of 0.1 g s-1 the test subject controlled the centrifugation himself and selected the acceleration values he was instructed to attain. The subjects were shown to be capable of assessing integrally the value of accelerations and estimating it with an error of 0.1-0.12 g. During an exposure to chest-to-back accelerations (+gx) limited to a value of 12 g and a gradient of 0.3 g s-1 the ability to select actively the rate of an acceleration increase was studied. Repeated centrifugations resulted in the formation and strengthening of the skill of maintaining a physiologically optimal rate of an acceleration increase. Onset rates were determined for every level of acceleration. When calculated as a mean, physiologically optimal values of the onset rate for 12 g were 0.16-0.18 g s-1. The findings give insight into important properties of the physiological structure of "gravipreferendum", which is the capability of man to differentiate between gravity levels and to select actively optimal rates of an increase in acceleration. PMID- 11913428 TI - Otolith functions in weightlessness. AB - The role of the vestibular organ in the exploration of space has been studied extensively during the past two decades. Many investigators have shown that some persons experience ill effects during the transition from the normal gravity to subgravity or weightlessness. Such adverse reactions can be related to a variety of sensory and somatic changes within the body systems; but it appears that the two major components of the unusual force field--namely, the absence of gravitational stimulation of the otolith organs and the occasional stimulation of the semicircular canals by head and body movements--bring about the motion sickness type reactions. Experiments in parabolic flights and in spacecraft revealed that the statolith organs respond to changes of acceleration during zero G. After an initial period of increased activity during the transition from 1 G to zero G, the number of nerve impulses from the otoliths is drastically decreased and later becomes steady on a somewhat lower than normal level of the discharge rate. The various theories concerning otolith responses in weightlessness are discussed and validated against the actual findings on astronauts and cosmonauts during spaceflight experiments and missions. PMID- 11913429 TI - Effect of 14 days of bed rest on urine metabolite excretion and plasma enzyme levels. AB - After one week of ambulatory baseline measurement, a group of eight men 19-26 years of age remained continuously recumbent for 14 days. Studies were continued for one week following the prolonged recumbency. Urine excretion rates for a number of constituents were determined 2 days before bed rest, on day 14 of bed rest, and day 6 after bed rest. Blood plasma samples were also obtained at these times, and analyzed for several enzymes. On day 14 of bed rest significant increases were observed in urine excretion of total osmotically active substances, magnesium, calcium, phosphate, creatinine, hydroxyproline, and 17-OH corticosteroids. A decrease occurred in urinary glucose excretion. Plasma levels of alkaline phosphatase and LDH-3 were depressed, while plasma GPT was elevated. Many of these changes persisted on day 6 after bed rest, and are interpreted as concomitants of the disuse atrophy of the musculoskeletal system that characterizes prolonged bed rest and weightlessness. PMID- 11913430 TI - Influence of simulated weightlessness on the rate of anomalies of the flour beetle Tribolium confusum. AB - Experiments with Tribolium confusum showed that the morphological characteristics of the beetles are modified by simulated weightlessness (fast running clinostat). Because of possible side effects due to differences in fertility of inbred lines, the first experiments were made with a genetically heterogeneous stock. Thereafter experiments were confirmed with inbred beetles. For both stocks a rise of mainly wing anomalies resulted from rotation of whole cultures of beetles within horizontal tubes. The extent to which these anomalies are teratogenetic or genetic has not yet been analysed in detail. PMID- 11913431 TI - The development of seedling shoots under space flight conditions. AB - The assumption that gravity is the major factor in the process of formation of plant polar axis was used as a working basis for the experiment. It was hypothesized by Merkys in 1973 that the effect of gravity related to axial polarity is similar to the process which determines the lateral polarity of shoots under the influence of gravity. There are two possibilities: (i) the development and morphogenesis of shoots takes place directly under the influence of gravity, or (ii) this process, at least during the first growth phases, is determined in the course of the germ development in the seed. In accordance with that assumption, the experiment was carried out in 1973. A special system was used for germinating and cultivating "Pioneer" and "Grybovsky rannyj" peas. The duration of the experiment under flight conditions was 48 hours in darkness, at 20 degrees C. The experimental conditions were the following: 1, the experiment in flight; 2, imitation of flight conditions using the horizontal clinostat; 3, vertical clinostat; 4, control (vertical plants). When the system was brought down to earth, the material was fixed and subjected to morphological and biochemical analysis. On the basis of the analysis, the following conclusion was drawn: during the first growth phases, the morphogenesis of shoots and roots apparently does not change under flight conditions. This conclusion was confirmed by planting those seedlings under earth conditions; normal plants were obtained whose growth and development were similar to the control seedlings. The problem of the influence of changed gravity, or the lack of it, on the growth and development of plants is discussed. The possible role of gravity on the formation of the polarity axes is also discussed from the point of view of generative development and the determination of some peculiarities of morphogenesis. PMID- 11913432 TI - Is the detection of optical activity in extraterrestrial samples a safe indicator for life? AB - Since Pasteur's times it has been accepted that optical activity and life are intimately correlated with each other, i.e. the one could not exist without the other. This argument logically allows the following speculation related to interplanetary exploration: detection of optical activity in an extraterrestrial sample would be the most straightforward proof of the existence of living matter outside the earth. On the other hand it has been shown that spontaneous production of optical activity can be achieved without the help of living matter, so that the close association of optical activity with life is not necessarily true. The paper discusses the possibilities which allow a proof of some sort of life on other planets by polarimetric methods. PMID- 11913433 TI - Privacy and the economics of personal health care information. PMID- 11913434 TI - Institutional review boards have new role in stem cell research. PMID- 11913436 TI - Vaccine from aborted fetus cell lines judged morally acceptable. PMID- 11913435 TI - Parents say federal agency helps protect researchers from more than their subjects. PMID- 11913437 TI - Candor and the court: the Supreme Court will confront as never before the violent nature of mid- and later-term abortion. PMID- 11913438 TI - The Genome Project: more than a medical milestone. PMID- 11913439 TI - The ethics and management of a Catholic hospital. PMID- 11913440 TI - Saying the unsaid: voicing quality-of-life criteria in an evangelical sanctity-of life principle. PMID- 11913441 TI - Child abuse or deep faith: medical neglect as a result of spiritual beliefs. PMID- 11913442 TI - Harvesting embryonic stem cells for research: response to NIH draft guidelines. PMID- 11913443 TI - State's partial-birth abortion ban supported. PMID- 11913445 TI - Abortion: a review of ethical aspects of public policy. PMID- 11913446 TI - Personhood, covenant, and abortion. PMID- 11913444 TI - Towards a theory of autonomy and informed consent. PMID- 11913447 TI - Confucian ethic of death with dignity and its contemporary relevance. AB - This paper advances three claims. First, according to contemporary Western advocates of physician-assisted suicide and voluntary euthanasia, "death with dignity" is understood negatively as bringing about death to avoid or prevent indignity, that is, to avoid a degrading existence. Second, there is a similar morally affirmative view on death with dignity in ancient China, in classical Confucianism in particular. Third, there is a consonance as well as dissonance between these two ethics of death with dignity, such as that the Confucian perspective would regard the argument for physician-assisted suicide and voluntary euthanasia as less than compelling because of the latter's impoverished vision of human life. PMID- 11913448 TI - Life sciences and space research. Volume IV. A session of the Sixth International Space Science Symposium, Mar del Plata, Argentina, May 11-19, 1965. PMID- 11913449 TI - Intracellular signalling involved in activation of the volume-sensitive K+ current in Ehrlich ascites tumour cells. AB - The cell swelling-activated K+ channel in Ehrlich ascites tumour cells has a conductance of 5 pS estimated from noise analysis of the volume-sensitive whole cell K+ current (I(K,vol)). I(K,vol) exhibits Goldman-Hodgkin-Katz type behaviour and is insensitive to clotrimazole, apamin and charybdotoxin (ChTX), but inhibited by clofilium. Its small conductance, lack of intrinsic voltage dependence and peculiar pharmacological profile are similar to properties described for the two-pore domain background K+ TASK channels. Neither Ca2+ nor ATP work as initiators in the activation of I(K,vol). In contrast, several investigations in Ehrlich cells suggest an important role for leukotriene D4 (LTD4) in the activation of I(K,vol). Under isotonic conditions, LTD4 activates Ca2+-dependent, ChTX-sensitive K+ channels as well as Ca2+-independent. ChTX insensitive K+ channels. The LTD4-activated, ChTX-insensitive K+ current exhibits a current-voltage relation, pharmacological profile and single channel conductance similar to that of I(K,vol), indicating that LTD4 is the signalling molecule responsible for activation of the volume-sensitive K+ channels in Ehrlich cells. Hypotonic swelling of Ehrlich cells results in translocation of the 85-kDa cytosolic (c) PLA2alpha to the nucleus where it is activated. This activation leads to an increase in arachidonic acid release followed by an increased release of leukotrienes, and is essential in cell swelling-induced activation of I(K,vol) and of the organic osmolyte channels. PMID- 11913451 TI - Apoptosis, cell volume regulation and volume-regulatory chloride channels. AB - Apoptosis occurs in response to various stimuli under physiological and pathological circumstances. A major hallmark of the programmed cell death is normotonic shrinkage of cells. Induction of the apoptotic volume decrease (AVD) was found to precede cytochrome c release, caspase-3 activation and DNA laddering. A broad-spectrum caspase inhibitor blocked these biochemical apoptotic events but failed to block the AVD. The normotonic AVD induction was coupled to facilitation of the regulatory volume decrease (RVD), which is attained by parallel operation of Cl- and K+ channels, under hypotonic conditions. Both the AVD induction and RVD facilitation were prevented by application of a blocker of volume-regulatory Cl- or K+ channels. Furthermore, apoptotic cell death was rescued by channel blocker-induced prevention of AVD. Thus, it is concluded that the AVD is produced under normotonic conditions by a mechanism similar, though without preceding swelling, to RVD and represents an early prerequisite to apoptotic events leading to cell death. It was previously reported that hypertonic stress triggers apoptosis in cell types that lack the regulatory volume increase (RVI) mechanism. Taken together, it is suggested that 'disordered' or altered cell volume regulation is associated with apoptosis. PMID- 11913450 TI - Serum- and glucocorticoid-dependent kinase, cell volume, and the regulation of epithelial transport. AB - Ample pharmacological evidence points to a role of kinases in the regulation of cell volume. Given the limited selectivity of most inhibitors, however, the specific molecules involved have remained largely elusive. The search for cell volume regulated genes in liver HepG2 cells led to the discovery of the human serum- and glucocorticoid-dependent serine/threonine kinase hsgk1. Transcription and expression of hsgk1 is markedly and rapidly upregulated by osmotic and isotonic cell shrinkage. The effect of osmotic cell shrinkage on hsgk1 is mediated by p38 kinase. Further stimuli of hsgk1 transcription include glucocorticoids, aldosterone, TGF-beta1, serum, increase of intracellular Ca2+ and phorbolesters, whereas cAMP downregulates hsgk1 transcription. The hsgk1 protein is expressed in several epithelial tissues including human pancreas, intestine, kidney, and shark rectal gland. Co-expression of hsgk1 with the renal epithelial Na+-channel ENaC or the Na+/K+/2Cl(-)-cotransporter NKCC2 (BSC1) in Xenopus oocytes, accelerates insertion of the transport proteins into the cell membrane and thus, stimulates channel or transport activity. Thus, hsgk1 participates in the regulation of transport by steroids and secretagogues increasing intracellular Ca2+-activity. The stimulation of hsgk1 transcription by TGF-beta1 may further bear pathophysiological relevance. PMID- 11913452 TI - The cytoskeleton and cell volume regulation. AB - Although the precise mechanisms have yet to be elucidated, early events in osmotic signal transduction may involve the clustering of cell surface receptors, initiating downstream signaling events such as assembly of focal adhesion complexes, and activation of, e.g. Rho family GTPases, phospholipases, lipid kinases, and tyrosine- and serine/threonine protein kinases. In the present paper, we briefly review recent evidence regarding the possible relation between such signaling events, the F-actin cytoskeleton, and volume-regulatory membrane transporters, focusing primarily on our own work in Ehrlich ascites tumer cells (EATC). In EATC, cell shrinkage is associated with an increase, and cell swelling with a decrease in F-actin content, respectively. The role of the F-actin cytoskeleton in cell volume regulation in various cell types has largely been investigated using cytochalasins to disrupt F-actin and highly varying effects have been reported. Findings in EATC show that the effect of cytochalasin treatment cannot always be assumed to be F-actin depolymerization, and that, moreover, there is no well-defined correlation between effects of cytochalasins on F-actin content and their effects on F-actin organization and cell morphology. At a concentration verified to depolymerize F-actin, cytochalasin B (CB), but not cytochalasin D (CD), inhibited the regulatory volume decrease (RVD) and regulatory volume increase (RVI) processes in EATC. This suggests that the effect of CB is related to an effect other than F-actin depolymerization, possibly its F actin severing activity. PMID- 11913453 TI - Necrotic volume increase and the early physiology of necrosis. AB - Whether a lethally injured mammalian cell undergoes necrosis or apoptosis may be determined by the early activation of specific ion channels at the cell surface. Apoptosis requires K+ and Cl- efflux, which leads to cell shrinking, an active phenomenon termed apoptotic volume decrease (AVD). In contrast, necrosis has been shown to require Na+ influx through membrane carriers and more recently through stress-activated non-selective cation channels (NSCCs). These ubiquitous channels are kept dormant in viable cells but become activated upon exposure to free radicals. The ensuing Na+ influx leads to cell swelling, an active response that may be termed necrotic volume increase (NVI). This review focuses on how AVD and NVI become conflicting forces at the beginning of cell injury, on the events that determine irreversibility and in particular, on the ion fluxes that decide whether a cell is to die by necrosis or by apoptosis. PMID- 11913454 TI - Cell cycle delay and apoptosis in response to osmotic stress. AB - As part of the urinary concentrating mechanism, renal inner medulla cells may be exposed to extremely variable NaCl and urea concentrations that can reach very high levels. A number of studies, reviewed herein, aim to understand how such osmotic stress affects the cells and what protective mechanisms might exist. The majority of these studies are done on inner medullary epithelial cells that grow continuously in tissue culture (mIMCD3). Cells grown at 300 mosmol/kg survive increase to 500 mosmol/kg by adding NaCl or urea, but only after a growth arrest of approximately 24 h. At a higher osmolality (650-700 mosmol/kg) most cells die within hours by apoptosis. The cells both in vitro and in vivo adapt to high osmolality by a number of mechanisms, including accumulation of variety of organic osmolytes and induction of heat shock proteins. The cell cycle delay results from blocks at the G1 and G2/M checkpoints and slowing during S. After adding NaCl, but not urea, the amount and transcriptional activity of p53 (the tumor suppressor protein) increases. The p53 is phosphorylated on ser-15 and is transcriptionally active at 500 mosmol/kg (associated with cell survival), but not at 700 mosmol/kg (associated with apoptosis). Reduction of p53 expression by p53 antisense oligonucleotide increases sensitivity of renal cells in culture to hyperosmotic stress caused by NaCl. The possible mechanisms of the protection action of p53 against hypertonic stress are discussed. PMID- 11913455 TI - Maintenance of genomic integrity in mammalian kidney cells exposed to hyperosmotic stress. AB - Changes in environmental salinity/osmolality impose an osmotic stress upon cells because, if left uncompensated, such changes will alter the conserved intracellular ionic milieu and macromolecular density, for which cell metabolism in most extant cells has been optimized. Cell responses to osmotic stress include rapid posttranslational and slower transcriptional events for the compensatory regulation of cell volume, intracellular electrolyte concentrations, and protein stability/activity. Changes in external osmolality are perceived by osmosensors that control the activation of signal transduction pathways giving rise to the above responses. We have recently shown that the targets of such pathways include cell cycle-regulatory and DNA damage-inducible genes (reviewed in Kultz, D., 2000. Environmental stressors and gene responses, Elsevier, Amsterdam. pp 157 179). Moreover, recent evidence suggests that hyperosmotic stress causes chromosomal aberrations and DNA double-strand breaks in mammalian cells. We propose that the modulation of cell cycle checkpoints and the preservation of genomic integrity are important aspects of cellular osmoprotection and as essential for cellular osmotic stress resistance as the capacity for cell volume regulation and maintaining inorganic ion homeostasis and protein stability/activity. PMID- 11913456 TI - Signaling and gene regulation by urea in cells of the mammalian kidney medulla. AB - Signaling by urea, although incompletely understood, is relevant both to cells of the mammalian kidney inner medulla and to all cells of the organism in the setting of advanced renal failure with its attendant accumulation of urea in the systemic circulation. The molecular events initiated by urea stress are distinct from those occurring in response to hypertonic stress; urea activates a characteristic subset of signaling events, which are in large part specific to cultured renal tubular epithelial cells. Interestingly, urea is protective of hypertonic NaCl-inducible apoptosis in this model. Details of this phenomenon are reviewed. The effect of urea has been likened to that of either hypertonicity or of a peptide mitogen. In preliminary expression array analyses, the profile of genes activated by urea stress in renal medullary cells, however, was found to be unique. PMID- 11913457 TI - Osmosensing and osmoregulatory compatible solute accumulation by bacteria. AB - Bacteria inhabit natural and artificial environments with diverse and fluctuating osmolalities, salinities and temperatures. Many maintain cytoplasmic hydration, growth and survival most effectively by accumulating kosmotropic organic solutes (compatible solutes) when medium osmolality is high or temperature is low (above freezing). They release these solutes into their environment when the medium osmolality drops. Solutes accumulate either by synthesis or by transport from the extracellular medium. Responses to growth in high osmolality medium, including biosynthetic accumulation of trehalose, also protect Salmonella typhimurium from heat shock. Osmotically regulated transporters and mechanosensitive channels modulate cytoplasmic solute levels in Bacillus subtilis, Corynebacterium glutamicum, Escherichia coli, Lactobacillus plantarum, Lactococcus lactis, Listeria monocytogenes and Salmonella typhimurium. Each organism harbours multiple osmoregulatory transporters with overlapping substrate specificities. Membrane proteins that can act as both osmosensors and osmoregulatory transporters have been identified (secondary transporters ProP of E. coli and BetP of C. glutamicum as well as ABC transporter OpuA of L. lactis). The molecular bases for the modulation of gene expression and transport activity by temperature and medium osmolality are under intensive investigation with emphasis on the role of the membrane as an antenna for osmo- and/or thermosensors. PMID- 11913458 TI - Changes in major intracellular osmolytes in L-929 cells following rapid and slow application of hyperosmotic media. AB - Cultured L-929 cells respond to media-made hyperosmotic (600 mOsmol/kg H2O) by addition of NaCl, sorbitol or proline by adjusting successively their intracellular level in different osmolytes: Na+, K+, amino acids and sorbitol. In the NaCl medium, Na+ and K+ are first to increase. Their concentration is then down-regulated while they are replaced by less disrupting osmolytes: amino acids and sorbitol. The amino-acid level is also adjusted with respect to the increase in sorbitol which starts only after 24 h, depending on the induction of aldose reductase. A similar evolution in the amount of these osmolytes is observed, with different time scales and amplitudes, depending on whether the osmotic shocks are applied abruptly or slowly, in a more physiological way. The interplay between the osmolytes is also different depending on their availability in the external medium. Such complex evolutions indicate that a cascade of interacting signals must be considered to account for the overall regulation process. It can hardly be fitted into a model implicating a single primary signalling event (early increase in ions or decrease in cell volume) as usually postulated. Also, the volume up-regulation is not significantly different in the different conditions, showing that it is not primarily dependent on the adjustment of the intracellular osmolarity which is reached immediately upon cell shrinkage and is maintained all over, independently of the availability and changes in nature of the osmolytes. PMID- 11913460 TI - Ion transport and ligand binding by the Na-K-Cl cotransporter, structure-function studies. AB - The cation-Cl cotransporters (CCCs) mediate the coupled movement of Na and/or K to that of Cl across the plasmalemma of animal cells. Eight CCCs have been identified to date: two Na-K-Cl cotransporters (NKCC), four K-Cl cotransporters (KCCs), one Na-Cl cotransporter (NCC) and one CCC interacting protein (CIP). All of the NKCCs and KCCs are inhibited by loop diuretics; mercury and other modifying agents are also known to block NKCC-mediated transport. In this work, we have utilized a mutational approach to study the interaction between different substrates and the NKCCs. We relied on the strategy of exchanging domains between functionally distinct carriers (the shark NKCCl and the human NKCCl) to identify residues or group of residues that are involved in the interaction with ions, loop diuretics and Hg. Our results show that the N- and C-termini have no role in determining the species differences in ion transport and bumetanide binding. On the other hand, the interaction between Hg and the NKCCs is found to partially involve the C-terminus through residues that contain available sulfhydryl groups. Within the transmembrane segments, variant residues in the 2nd, 4th and 7th predicted alpha-helices are shown to encode the differences in ion transport between the shark and the human cotransporters. For loop diuretic binding, several regions throughout the central domain appear to be involved. Interestingly, these regions are not the same as those involved in cation or anion transport, and in Hg binding. PMID- 11913459 TI - Some ways of looking at compensatory kosmotropes and different water environments. AB - Hydration of macromolecular structures determines biological activity. Stabilizing solutes are kosmotropic (increase order of water) rather than chaotropic (decrease order). Preferential hydration of surfaces is a thermodynamic consequence of the solution behavior of kosmotropic solutes, but inconsistencies imply interactions such as the hydration of specific sites within macromolecules. Thermodynamic measures require bulk pure solutes; here simpler measures of the effects on bulk water, water at surfaces and hydration water of probes have been applied to solutes including natural stabilizers, analogues and example chaotropes. Changes in the near-infrared spectra, water proton NMR chemical shifts and relaxation times measure changes in the bulk liquid; HPLC column retention of solutes indicate interactions with hydration water at different surfaces, and fluorescence probes detect effects on functional group hydration water. Ab initio calculations and Monte-Carlo simulations of the solutes in water measure the energetics of the solute-water interactions, the dipole moments of these molecules, their charge distributions and the effect of the solute molecules on the structure of water. The rankings of the test solutes by these measures are not consistent. Thus, stabilizing solutes are not interchangeable in biological systems and the intracellular replacement of one by another could affect the integration of cell metabolism. PMID- 11913461 TI - K-Cl co-transport: immunocytochemical and functional evidence for more than one KCC isoform in high K and low K sheep erythrocytes. AB - K-Cl co-transport (COT) is significantly higher in low K (LK), L-antigen (L) positive, than in high K (HK), M-antigen (M) positive, sheep red blood cells (SRBCs) and is inhibited by sheep allo-anti-L1 antibody. To answer the question of whether this difference in K-Cl co-transport activity resides at the level of the transporter or its regulation, a combined immunocytochemical and functional approach was taken. At least four isoforms of K-Cl COT encoded by different KCC genes are known, with 12 transmembrane domains and cytoplasmic C- and N-terminal domains (Ctd and Ntd, respectively). Polyclonal anti-rat (rt)KCC1 antibodies against a fusion peptide with 77 amino acids from the Ctd of rtKCC1 and anti human (h)KCC3 against an 18-aa peptide from the Ntd of hKCC3, were prepared in rabbits (rb). Two distinctly separate protein bands of 180 and 145 kDa molecular mass were detected in hemoglobin-free ghosts from RBCs of two LK (one homozygous LL and one heterozygous LM) and one HK (homozygous MM) sheep by Western blots with rb anti-rtKCC1 and rb anti-hKCC3. Confocal microscopy showed specific immunostaining of KCC1 with rb anti-rtKCC1, and of KCC3 with rb anti-hKCC3, in white ghosts from both LK and HK SRBCs. To test the functional heterogeneity of K Cl COT, the effect of the anti-L1 antibody was assessed on K-Cl COT activated by the kinase inhibitor staurosporine. Incubation of LK SRBCs with anti-L1 serum inhibited by 30% staurosporine-stimulated K-Cl COT suggesting that approximately two-thirds of the transport activity is independent of the L1 antigen. That staurosporine altered the L1 antigen/antibody reaction is unlikely since the action of another antibody, anti-Lp, stimulating the Na/K pump flux, was not modified. The present results, in conjunction with earlier work, lead to the hypothesis that the partial anti-L1 inhibition of K-Cl COT may be related to the molecular KCC dimorphism, seen in these cells with anti-KCC1 and anti-KCC3 antibodies. PMID- 11913462 TI - Cl(-)-ATPases: biological active transporters. AB - Five widely documented mechanisms of chloride transport across plasma membranes are: anion-coupled antiport; sodium and hydrogen-coupled symport; Cl- channels; and an electrochemical coupling process. No genetic evidence has yet been provided for primary active chloride transport despite numerous reports of cellular Cl(-)-stimulated ATPases co-existing, in the same tissue, with uphill chloride transport that could not be accounted for by the five common chloride transport processes. Cl(-)-stimulated ATPase activity is a common property of practically all biological cells with the major location being of mitochondrial origin. It also appears that plasma membranes are sites of Cl(-)-stimulated ATPase activity. Recent studies of Cl(-)-stimulated ATPase activity and active chloride transport in the same membrane system, including liposomes, suggest a mediation by the ATPase in net movement of chloride up its electrochemical gradient across plasma membranes. Further studies, especially from a molecular biological perspective, are required to confirm a direct transport role to plasma membrane-localized Cl(-)-stimulated ATPases. PMID- 11913463 TI - Different activation mechanisms of cystic fibrosis transmembrane conductance regulator expressed in Xenopus laevis oocytes. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP sensitive Cl- channel that is defective in cystic fibrosis (CF). The most frequent mutation, namely deltaF508-CFTR, accounts for 66% of CF. Here we show that cAMP-activation of CFTR occurs via at least two distinct pathways: activation of CFTR molecules already present in the plasma membrane and protein kinase A (PKA)-mediated vesicular transport of new CFTR molecules to the plasma membrane and functional insertion into the membrane. We investigated the mechanisms that are responsible for these activation pathways using the Xenopus laevis oocytes expression system. We expressed CFTR and recorded continuously membrane current (Im), conductance (Gm) and capacitance (Cm), which is a direct measure of membrane surface area. Expression of CFTR alone did not change the plasma membrane surface area. However, activation of CFTR with cAMP increased Im, Gm and Cm while deltaF508-CFTR-expressing oocytes showed no response on cAMP. Inhibition of protein kinase A or buffering intracellular Ca2+ abolished the cAMP induced increase in Cm while increases of Im and Gm were still present. ATP or the xanthine derivative 8-cyclopentyl-1,3-dipropylxanthine (CPX) did not further activate CFTR. Insertion of pre-formed CFTR into the plasma membrane could be prevented by compounds that interfere with intracellular transport mechanisms such as primaquine, brefeldin A, nocodazole. From these data we conclude that cAMP activates CFTR by at least two distinct pathways: activation of CFTR already present in the plasma membrane and exocytotic delivery of new CFTR molecules to the oocyte membrane and functional insertion into it. PMID- 11913464 TI - Osmolality influences bistability of membrane potential under hypokalemic conditions in mouse skeletal muscle: an experimental and theoretical study. AB - The membrane potential in mouse skeletal muscle depends on both extracellular osmolality and potassium concentration. These dependencies have been related to two membrane transporters, Na+/K+/2Cl- co-transporter and the inward potassium rectifier channel. To investigate the relation of the Na+/K+/2Cl- co-transporter and the inward potassium rectifier channel in a qualitative way, a combined electrophysiological and modelling approach was used. The experimental results show that the bistability of the membrane potential, which is related to the conductive state of the inward potassium rectifier channel, is shifted to higher extracellular potassium values when medium osmolality is increased. These results are confirmed by the computer simulation calculations for increased co transporter flux. The combined results indicate that the co-transporter is capable of modulating the conductive state of the inward potassium rectifier channel. PMID- 11913465 TI - Expression of cystic fibrosis transmembrane conductance regulator in the skin of the toad, Bufo bufo and possible role for Cl- transport across the heterocellular epithelium. AB - Evidence is discussed that apical CFTR Cl- channels of mitochondria-rich (MR) cells of Bufo bufo skin conduct beta-adrenergic receptor-activated Cl- currents. Ussing chambers studies revealed the following selectivity sequence of the receptor activated conductance, Cl- > Br- > NO3- > I-. With ion selective microelectrode-techniques, it was shown that receptor-coupled Cl- channels are not located in principal cells. A small conductance (7-10 pS) CFTR-like Cl- channel is located in the apical plasma membrane of MR cells. Short life times of sealed patches prevented detailed study of its selectivity to other halide ions and its molecular regulation. With monoclonal hCFTR-antibodies, selective expression in MR cells of the targeted antigens could be demonstrated. A transcript of CFTR was amplified in the skin, and a bbCFTR cDNA clone was generated from toad skin mRNA that exhibits 89% amino acid identity with the human homologue. The frequency of obtaining channels in patch clamp studies was too low for accounting quantitatively for the macroscopic conductance. Since MR cells were isolated by trypsin, and a putative extracellular loop of the deduced bbCFTR protein contains a target peptide bond for trypsin, enzyme treatment may have destroyed apical CFTR molecules. PMID- 11913466 TI - Molecular physiology of osmoregulation in eels and other teleosts: the role of transporter isoforms and gene duplication. AB - This review focuses on recent developments in the molecular biology of ion and water transporter genes in fish and the potential role of their products in osmoregulation in both freshwater and seawater environments. In particular details of isoforms of various ATPases, co-transporters, exchangers and ion channels in the eel as well as other teleost species are described. Many of the teleost transporter isoforms discovered so far, appear to occur as twin or duplicate copies compared to their homologous counterparts in higher vertebrates, although these duplicate isoforms often have distinct tissue-specific and developmental stage-dependent expression patterns. The possible meaning of this information will be examined in relation to the fish genome duplication debate. PMID- 11913468 TI - Lysophosphatidylcholine-induced taurine release in HeLa cells involves protein kinase activity. AB - It has recently been demonstrated that exogenous addition of low concentrations (< 15 microM) of lysophosphatidyl choline (LPC, palmitic acid in the sn-1 position) induces a transient increase in taurine efflux from HeLa cells in a process that seems to involve generation of reactive oxygen species (ROS) and tyrosine phosphorylation (J. Membrane Biol. 176 (2000) 175-185). We now demonstrate that LPC also induces release of taurine under isotonic conditions in mouse fibroblast (NIH/3T3) and Ehrlich ascites tumor cells. Furthermore, we show that in the case of HeLa cells addition of the calmodulin antagonist W-7 (50 microM) or the calmodulin-dependent kinase II (CaMKII) inhibitor KN-62 (10 microM) reduces the LPC-induced taurine release under isotonic conditions. Conversely, addition of a standard protein kinase C (PKC) inhibitor chelerythrine (10 microM) leads to a potentiation of the LPC-induced taurine efflux, whereas direct activation of PKC by the phorbol ester PMA has no effect. It is suggested that the putative generation of ROS following addition of LPC is modulated by calmodulin/CaMKII, and that the effect of chelerythrine is more likely related to the ROS production than to PKC inhibition. PMID- 11913467 TI - K+ conductance activated during regulatory volume decrease. The channels in Ehrlich cells and their possible molecular counterpart. AB - K+ currents activated by hypotonic cell swelling have been studied in Ehrlich ascites tumour cells by the whole-cell recording mode of the patch-clamp technique. K+ together with Cl- currents developed in the absence of added intracellular Ca2+ and with strong buffering of internal Ca2+ in experiments conducted at 37 degrees C. Manipulation of the extracellular medium with other cations suggests a selectivity sequence of K+ > Rb+ > NH4+ > or = Na+ approximately equals Li+ approximately equals Cs+. The current-voltage relationship of the volume-sensitive K+ current was well fitted with the Goldman Hodgkin-Katz current equation between -130 and 20 mV at both physiological and high K+ extracellular solutions. The class III antiarrhytmic drug clofilium blocked the volume-sensitive K+ current in a voltage-independent manner. Clofilium was also found to be a strong inhibitor of the regulatory volume decrease (RVD) response of Ehrlich cells. The leukotriene D4 (LTD4) can activate the same current in isotonicity, consistent with a role for this compound in the signalling process of volume regulation. It is suggested that K+ channels activated by cell swelling belong to the so-called background K+ channel group. These are voltage-independent channels which underlie the resting potential of many cells and have recently been identified as belonging to a family of K+ channels with two pore domains in tandem (2P-4TM). Preliminary experiments show the presence of the TASK-2 channel, a member of the 2P-4TM family inhibited by acid extracellular pH, in Ehrlich cells and suggest that it might underlie the swelling-induced K+ current. PMID- 11913469 TI - D-glucose transport in decapod crustacean hepatopancreas. AB - Physiological mechanisms of gastrointestinal absorption of organic solutes among crustaceans remain severely underinvestigated, in spite of the considerable relevance of characterizing the routes of nutrient absorption for both nutritional purposes and formulation of balanced diets in aquaculture. Several lines of evidence attribute a primary absorptive role to the digestive gland (hepatopancreas) and a secondary role to the midgut (intestine). Among absorbed organic solutes, the importance of D-glucose in crustacean metabolism is paramount. Its plasma levels are finely tuned by hormones (crustacean hyperglycemic hormone, insulin-like peptides and insulin-like growth factors) and the function of certain organs (i.e. brain and muscle) largely depends on a balanced D-glucose supply. In the last few decades, D-glucose absorptive processes of the gastrointestinal tract of crustaceans have been described and transport mechanisms investigated, but not fully disclosed. We briefly review our present knowledge of D-glucose transport processes in the crustacean hepatopancreas. A discussion of previous results from experiments with hepatopancreatic epithelial brush-border membrane vesicles is presented. In addition, recent advances in our understandings of hepatopancreatic D-glucose transport are shown, as obtained (1) after isolation of purified R-, F-, B- and E cell suspensions from the whole organ by centrifugal elutriation, and (2) by protein expression in hepatopancreatic mRNA-injected Xenopus laevis oocytes. In a perspective, the applicability of these novel methods to the study of hepatopancreatic absorptive function will certainly improve our knowledge of this structurally complex organ. PMID- 11913470 TI - Simultaneous measurements of calcium mobilization and afferent nerve activity in electroreceptor organs of anesthetized Kryptopterus bicirrhis. AB - The transduction pathway of ampullary electroreceptor organs involves ionic currents. It has been shown that calcium, as well as sodium and potassium play important parts in this process. In this study we examine the stimulus-evoked changes in the Fura-2 ratio in electroreceptor cells. Furthermore, we recorded stimulus-evoked Fura-2 ratio changes while Na+ and K+ channels were blocked by amiloride and TEA. Simultaneously, extracellular recordings of the afferent spike activity were made. The results show the presence of stimulus evoked fluctuations in the Fura-2 ratio. These fluctuations can be abolished by the application of Cd2+, TEA, and amiloride. The stimulus-evoked activity of the afferent nerve was decreased due to application of these drugs. We conclude that the transduction current is carried by Na+, K+, and probably Ca2+. This fits the existing model on transduction in electroreceptors. PMID- 11913471 TI - Should statin therapy be considered for patients with elevated C-reactive protein? The need for a definitive clinical trial. PMID- 11913472 TI - Can metabolic manipulation reverse myocardial dysfunction? PMID- 11913473 TI - Diagnosing myocardial iron overload. PMID- 11913474 TI - Dofetilide: what role in the treatment of ventricular tachyarrhythmias? PMID- 11913475 TI - Circadian variation of arrhythmic events, electrophysiological properties, and the autonomic nervous system. PMID- 11913476 TI - Vasodilator pre-treatment of human radial arteries. PMID- 11913477 TI - New approaches to antiarrhythmic therapy; emerging therapeutic applications of the cell biology of cardiac arrhythmias. AB - Cardiac arrhythmias complicate many diseases affecting the heart and the circulation, and incorporate a multiplicity of underlying mechanisms. The evolution of scientific knowledge has made the complex changes produced by cardiovascular disease sufficiently understood at the organ, cellular, and molecular levels such that there is a diversity of therapeutic targets for pharmacological therapy and/or prevention. Moreover, the approach of rational drug design, in mechanism- and disease-specific fashion, facilitates targeting of therapy via molecular, structural and translational biology. Additional approaches, employing similar drug-design strategies but based on gene therapy and transcriptional and translational modification are on the horizon. Hence, there is reason to be optimistic regarding the design, testing and clinical availability of novel antiarrhythmic therapies. PMID- 11913478 TI - Effects of trimetazidine on the contractile response of chronically dysfunctional myocardium to low-dose dobutamine in ischaemic cardiomyopathy. AB - BACKGROUND: There is evidence that trimetazidine, an anti-ischaemic agent with a direct cytoprotective effect on the myocardium, is effective in stable angina. However, it is not clear whether trimetazidine can improve the mechanical efficiency of chronically dysfunctional myocardium, and whether this potentially beneficial effect can translate into improvements in left ventricular function as well as functional capacity. METHODS: Thirty-eight patients (52.7 +/- 8 years) with post-necrotic left ventricular dysfunction (ejection fraction: 33 +/- 5%) and multivessel coronary artery disease were studied. Patients were randomized into two matched groups. Group A received trimetazidine (20 mg three times daily) for 2 months, while group B received a placebo during the same period. The usual antianginal medications were not altered during the study. At baseline and after 2 months, all patients underwent low-dose dobutamine echocardiography (5-20 microg x kg(-1) x min(-1)), and a symptom-limited cardiopulmonary exercise test. RESULTS: On initial evaluation, systolic wall thickening score index, heart rate, systolic blood pressure and rate pressure product were similar at rest and peak dobutamine in both groups. However, at 2 months, group A patients had significant improvements in the rest and peak systolic wall thickening score index (13% and 20.7%, P<0.001) and ejection fraction (19.7% and 14.1%, P<0.001) without concomitant changes in heart rate and blood pressure. Peak VO2 was also significantly increased in patients taking trimetazidine (15%, P=0.001 vs controls). CONCLUSIONS: In patients with ischaemic cardiomyopathy, trimetazidine improves the contractile response of chronically dysfunctional myocardium to dobutamine without haemodynamic changes. This effect was associated with improvements in left ventricular function and peak VO2. PMID- 11913479 TI - Cardiovascular T2-star (T2*) magnetic resonance for the early diagnosis of myocardial iron overload. AB - AIMS: To develop and validate a non-invasive method for measuring myocardial iron in order to allow diagnosis and treatment before overt cardiomyopathy and failure develops. METHODS AND RESULTS: We have developed a new magnetic resonance T2-star (T2*) technique for the measurement of tissue iron, with validation to chemical estimation of iron in patients undergoing liver biopsy. To assess the clinical value of this technique, we subsequently correlated myocardial iron measured by this T2* technique with ventricular function in 106 patients with thalassaemia major. There was a significant, curvilinear, inverse correlation between iron concentration by biopsy and liver T2* (r=0.93, P<0.0001). Inter-study cardiac reproducibility was 5.0%. As myocardial iron increased, there was a progressive decline in ejection fraction (r=0.61, P<0.001). All patients with ventricular dysfunction had a myocardial T2* of <20 ms. There was no significant correlation between myocardial T2* and the conventional parameters of iron status, serum ferritin and liver iron. Multivariate analysis of clinical parameters to predict the requirement for cardiac medication identified myocardial T2* as the most significant variable (odds ratio 0.79, P<0.002). CONCLUSIONS: Myocardial iron deposition can be reproducibly quantified using myocardial T2* and this is the most significant variable for predicting the need for ventricular dysfunction treatment. Myocardial iron content cannot be predicted from serum ferritin or liver iron, and conventional assessments of cardiac function can only detect those with advanced disease. Early intensification of iron chelation therapy, guided by this technique, should reduce mortality from this reversible cardiomyopathy. PMID- 11913480 TI - A multicentre, double-blind randomized crossover comparative study on the efficacy and safety of dofetilide vs sotalol in patients with inducible sustained ventricular tachycardia and ischaemic heart disease. AB - BACKGROUND: Antiarrhythmic drugs are still used for the treatment of ventricular tachyarrhythmias, in combination with implantable cardioverter-defibrillators or without them. AIM OF THE STUDY: In a double-blind randomized crossover design, the short- and long-term efficacy and safety of oral dofetilide or oral sotalol were compared in 135 patients with ischaemic heart disease and inducible sustained ventricular tachycardia. METHODS: The inducibility of ventricular tachycardia was determined by programmed electrophysiological stimulation at baseline. Patients were then blindly randomized to receive either oral dofetilide 500 microg twice daily or oral sotalol 160 mg twice daily, for 3 to 5 days. Suppression of inducible ventricular tachycardia on the drug was then assessed by programmed electrophysiological stimulation. After a wash-out period of at least 2.5 days, the patients received the alternative treatment for 3 to 5 days. Suppression of inducible ventricular tachycardia on the alternate drug was again determined by programmed electrophysiological stimulation. Selection of long-term treatment was allocated blindly according to programmed electrophysiological stimulation results. RESULTS: During the acute phase, 128 patients received both dofetilide and sotalol. Sixty-seven patients were responders to either drug. Forty-six patients (35.9%) were responders to dofetilide compared with 43 (33.6%) to sotalol (P=ns). Only 23 patients responded to both dofetilide and sotalol. Adverse events, deemed to be treatment related, were seen in 2.3% of patients receiving dofetilide and 8.6% of patients receiving sotalol (P=0.016). Three patients on dofetilide had torsade de pointes. Two patients receiving sotalol died during the acute phase (one was arrhythmic death, and the other was due to heart failure). During the long-term phase, two of 42 patients (4.8%) receiving dofetilide and three of 27 patients (11.1%) receiving sotalol withdrew from treatment due to lack of efficacy. Overall, during the long-term phase, 23.8% of the patients receiving dofetilide and 37.0% of the patients receiving sotalol, withdrew from treatment with a similar pattern of withdrawals for the two drugs. CONCLUSION: Dofetilide was as efficacious as sotalol in preventing the induction of sustained ventricular tachycardia. There was no concordance in the response rate in two-thirds of the patients. Dofetilide was significantly better tolerated during the acute phase than sotalol. Both dofetilide and sotalol were well tolerated during the long term with no statistically significant difference in the adverse events. PMID- 11913481 TI - Atrial and ventricular refractoriness in paced patients; circadian variation and its relationship to autonomic nervous system activity. AB - AIMS: To examine whether atrial and ventricular effective refractory periods exhibit circadian variation and whether the latter is correlated with fluctuations in autonomic nervous system tone. METHODS AND RESULTS: We studied 24 patients aged 67.1 +/- 9.6 years, 11 of whom were paced for complete heart block and 13 for sick sinus syndrome. Atrial and ventricular effective refractory periods were measured bihourly over a 24-h period, using the pacemaker programming capabilities, at basic cycle lengths of 600 ms and 500 ms. During the same time period we evaluated autonomic nervous system activity in patients paced for complete heart block, expressed by spectral power indexes in low frequency and high frequency areas of heart rate variability. Atrial and ventricular effective refractory periods showed significant circadian variation at both basic cycle lengths, with the highest values occurring between 22:00 and 06:00. At times, the atrial and ventricular effective refractory periods of the patients with sick sinus syndrome differed significantly from those with complete heart block. Furthermore, atrial and ventricular effective refractory periods in patients with complete heart block exhibited a strong negative correlation with the low frequency/high frequency ratio. CONCLUSION: Our data show that atrial and ventricular effective refractory periods in DDD paced patients exhibit significant circadian variation that is strongly correlated with variations in autonomic nervous system activity in patients with complete heart block. PMID- 11913482 TI - Clinical value of left atrial appendage flow velocity for predicting of cardioversion success in patients with non-valvular atrial fibrillation. AB - BACKGROUND: Echocardiographic parameters for predicting cardioversion outcome in patients with non-valvular atrial fibrillation are not accurately defined. OBJECTIVE: To evaluate the role of left atrial appendage flow velocity detected by transoesophageal echocardiography for prediction of cardioversion outcome in patients with non-valvular atrial fibrillation enrolled in a prospective. multicentre, international study. METHODS: Four hundred and eight patients (257 males, mean age: 66 +/- 10 years) with non-valvular atrial fibrillation lasting more than 48 h but less than 1 year underwent transthoracic echocardiography and transoesophageal echocardiography before either electrical (n=324) or pharmacological (n=84) cardioversion. RESULTS: Cardioversion was successful in restoring sinus rhythm in 328 (80%) and unsuccessful in 80 patients (20%). Mean left atrial appendage peak emptying flow velocity was significantly higher in patients with successful than in those with unsuccessful cardioversion (32.4 +/- 17.7 vs 23.5 +/- 13.6 cm x s(-1); P<0.0001). At multivariate logistic regression analysis, three parameters proved to be independent predictors of cardioversion success: the atrial fibrillation duration <2 weeks (P=0.011, OR=4.9, CI 95%=1.9 12.7), the mean left atrial appendage flow velocity >31 cm x s(-1) (P=0.0013, OR=2.8, CI 95%=1.5-5.4) and the left atrial diameter <47 mm (P=0.093, OR=2.0, CI 95%=1.2-3.4). These independent predictors of cardioversion success outperformed other univariate predictors such as left ventricular end-diastolic diameter <58 mm, ejection fraction >56% and the absence of left atrial spontaneous echo contrast. CONCLUSION: In patients with non-valvular atrial fibrillation, measurement of the left atrial appendage flow velocity profile by transoesophageal echocardiography before cardioversion provides valuable information for prediction of cardioversion outcome. PMID- 11913483 TI - Vasodilator pre-treatment of human radial arteries; comparison of effects of phenoxybenzamine vs papaverine on norepinephrine-induced contraction in vitro. AB - AIMS: The radial artery, increasingly used for coronary artery bypass grafting (CABG). has a potential for spasm which may increase peri-operative risk. Increased alpha-adrenoceptor activation is a key candidate for the spasm. We studied the effects of vasoconstriction in a radial artery, which had undergone brief exposure to the alpha-adrenoceptor antagonist phenoxybenzamine vs the opioid derivative papaverine. METHODS AND RESULTS: Using standard classical organ bath techniques, concentration responses were obtained to norepinephrine in segments of radial artery from 12 CABG patients pre- and post-incubation for 20 min in either phenoxybenzamine 10(-6) M or papaverine 3 x 10(-3) M. Responses were reassessed 2, 4 and 18 h after washout of phenoxybenzamine and 2, 4, 8 and 18 h after washout of papaverine. There was concentration-dependent constriction to norepinephrine (maximum response 0.89 +/- 0.20 (SEM) g x mm(-1), n=6). Constriction to norepinephrine was abolished immediately after incubation in phenoxybenzamine and remained completely inhibited for at least 18 h (P<0.0001 ANOVA phenoxybenzamine pre-treated vs controls). Most of the inhibition of concentration-dependent constriction to norepinephrine following pre-treatment with papaverine was lost 8 h later. CONCLUSION: Radial artery vasoconstriction induced by a clinically relevant agonist, norepinephrine, may be prevented for at least 18 h by pre-incubation in phenoxybenzamine, in contrast to the brief inhibition achieved by pre-treatment with papaverine. Adding phenoxybenzamine to radial artery graft bathing solution may improve early outcome following CABG. PMID- 11913484 TI - Healthy ageing: addressing acute coronary syndrome? AB - Acute coronary syndromes contribute significantly to the patterns of morbidity and mortality in the elderly. A meta-analysis of the pathology of acute myocardial infarction has shown that 80-90% of episodes result from the rupture of small, unstable lesions that cause <70% diameter stenosis. Statins have been shown to stabilize the architecture of atherosclerotic plaques in humans and in animals. Statin treatment has also been shown to restore endothelial function, inhibit platelet thrombus formation and exert an anti-inflammatory effect. Collectively, these data support the use of statin therapy in those at risk for acute coronary syndromes, of whom the elderly are foremost. PMID- 11913485 TI - Dementia at old age: a clinical end-point of atherosclerotic disease. AB - The main causes of dementia at old age are Alzheimer's disease and vascular dementia. The clinical presentations of late-onset Alzheimer's disease and vascular dementia can barely be differentiated. This clinical observation is supported by pathological findings. Late-onset dementia should be considered a multifactorial disease, in which both vascular factors and amyloid dispositions contribute to cognitive decline. PMID- 11913486 TI - Unhealthy ageing: functional and socioeconomic impact. AB - The long-term impacts of disease on functional status and on cost loom larger in the elderly than in any other age group. In older patients, the chronic sequelae of myocardial infarction and stroke often account for greater functional disability and resource utilization than does the initial acute presentation. In order to study this, quantitative measures from clinical and functional status assessments must be related to lifelong resource use. Rigorous measurement of such outcomes can render it possible to measure the impact on the health care system of specific cardiovascular and cerebrovascular consequences of 'unhealthy ageing'. If performed in the context of a randomized clinical trial studying the effect of preventive measures, this approach can also lay the groundwork for assessment of the clinical and economic benefits of measures taken to reduce such morbidity. PMID- 11913487 TI - Healthy ageing: ageing safely. AB - The population of the developed world is steadily ageing. In the European Union, approximately 22% of persons are over 60 years of age and this is projected to increase to more than 27% by the year 2020. This has major implications for health care resources and the productive workforce. Ageing is accompanied by a decline in the physiological reserve of all organ systems, compromising homeostasis and resistance to disease. Thus, when disease develops in the elderly it has an increased impact on organ systems not directly involved. This places older people at risk for multiple simultaneous pathologies. Treatment often requires polypharmacy, which often is accompanied by drug interactions and adverse reactions. The pattern of sequential and comorbid disease often means that the later years of life are associated with an accumulating toll of disability, which in turn consumes a high proportion of health-care resources. The major goal of health care in the elderly should be to compress morbidity into the end of the normal lifespan. To achieve this, it will be necessary to redefine our approaches to treatment in the elderly and to develop an evidence base to inform this process. PMID- 11913488 TI - The use of growth factors in the proliferation of avian articular chondrocytes in a serum-free culture system. AB - The purpose of this research was to develop a serum-free culture system for the proliferation of articular chondrocytes. Various growth factors and hormones were tested for their ability to stimulate avian articular chondrocyte proliferation in a defined, serum-free media. Multiple members of the fibroblast growth factor (FGF) family (FGFs: 2, 4, and 9), insulin-like growth factor-1 (IGF-1) and transforming growth factor beta (TGF-beta) significantly stimulated H-thymidine uptake by chondrocytes grown in an adherent serum-free, culture system. Double or triple combinations of these mitogenic growth factors further stimulated cell proliferation to levels that were equivalent to, or surpassed those of cells grown in serum. Although proliferation was maximally stimulated, chondrocytes grown in the presence of FGF-2, IGF-1, and TGF-beta, began to exhibit changes in morphology and collagen II expression declined. This culture system could be used to rapidly expand a population of articular chondrocytes prior to transferring these cells to a non-adherent culture system, which could then stabilize the chondrocyte phenotype and maximize matrix synthesis and integrity. PMID- 11913489 TI - GDF-5 deficiency in mice leads to disruption of tail tendon form and function. AB - Although the biological factors which regulate tendon homeostasis are poorly understood, recent evidence suggests that Growth and Differentiation Factor-5 (GDF-5) may play a role in this important process. The purpose of this study was to investigate the effect of GDF-5 deficiency on mouse tail tendon using the brachypodism mouse model. We hypothesized that GDF-5 deficient tail tendon would exhibit altered composition, ultrastructure, and biomechanical behavior when compared to heterozygous control littermates. Mutant tail tendons did not display any compositional differences in sulfated glycosaminoglycans (GAG/DNA), collagen (hydroxyproline/DNA), or levels of fibromodulin, decorin, or lumican. However, GDF-5 deficiency did result in a 17% increase in the proportion of medium diameter (100-225 nm) collagen fibrils in tail tendon (at the expense of larger fibrils) when compared to controls (p < 0.05). Also, mutants exhibited a trend toward an increase in irregularly-shaped polymorphic fibrils (33% more, p > 0.05). While GDF-5 deficient tendon fascicles did not demonstrate any significant differences in quasistatic biomechanical properties, mutant fascicles relaxed 11% more slowly than control tendons during time-dependent stress-relaxation tests (p < 0.05). We hypothesize that this subtle alteration in time-dependent mechanical behavior is most-likely due to the increased prevalence of irregularly shaped type I collagen fibrils in the mutant tail tendons. These findings provide additional evidence to support the conclusion that GDF-5 may play a role in tendon homeostasis in mice. PMID- 11913491 TI - Cell cluster formation in degenerate lumbar intervertebral discs is associated with increased disc cell proliferation. AB - Healthy human intervertebral discs contain relatively few cells and these are sparsely distributed. A characteristic feature of disc degeneration, however, is the appearance of cell clusters, particularly in damaged areas. How these clusters form is currently unknown. We have examined excised pathological human discs for evidence of cell proliferation. Disc sections were immunostained for the proliferating cell nuclear antigen (PCNA) and the proliferation-associated Ki 67 antigen. PCNA immunopositive cells were observed within degenerate discs, commonly though not exclusively, in cell clusters. Cells immunopositive for the Ki-67 antigen were less prevalent than those for PCNA, but similarly were observed frequently within clusters in degenerate discs. In contrast, immunopositivity for these markers was not common in less degenerate discs or in areas of the disc where cell clusters were not observed. These observations suggest that disc cell proliferation is associated with disc degeneration and is the likely cause of cell cluster formation. PMID- 11913490 TI - Proinflammatory cytokines regulate the gene expression of hyaluronic acid synthetase in cultured rabbit synovial membrane cells. AB - To elucidate the mechanism of accumulation and fragmentation of hyaluronic acid (HA) under inflammatory conditions, we investigated the effect of proinflammatory cytokines on hyaluronic acid synthetase (HAS) mRNA expression using cultured rabbit synovial membrane cells. HASs mRNA levels were determined by real-time PCR. HAS2 mRNA expression was maximally enhanced 3.3- and 2.8-fold after 3-hour stimulation with IL-1beta (1 ng/ml) and after 1-hour stimulation with TNF-alpha (10 ng/ml). HAS3 mRNA expression was increased by a maximum of 4.3 times after 3 hour stimulation with IL-1beta (10 ng/ml), whereas 1-hour stimulation with TNF alpha (10 ng/ml) and IFN-gamma (10 ng/ml) induced around a 2.5-fold increase in HAS3 mRNA. Although IFN-gamma (1-100 ng/ml) alone showed little effect on HAS2 mRNA expression, the effect was synergized by combined with both IL-1beta and TNF alpha, substantially increasing HAS2 mRNA expression. These results suggest that proinflammatory cytokines regulate the HAS expression, and consequently may contribute to the accumulation and fragmentation of HA. PMID- 11913492 TI - Structural varieties of small proteoglycans in human spinal ligament. AB - Three types of small proteoglycan were purified from human spinal ligaments by ultracentrifugation, ion-exchange chromatography, gel-chromatography, and hydrophobic chromatography. Two of them were identified as decorin and biglycan, and the other was thought to be a decorin-subtype. Molecular sizes of decorin and decorin-subtype were both 85 kDa, and that of biglycan was 200 kDa. N-Terminal amino acid sequence of decorin-subtype corresponded with that of decorin, although it was different from decorin in terms of composition of amino acids and glycosaminoglycan chains, and reactivity with anti-human decorin antibody. The ratios of chondroitin sulfate to dermatan sulfate contained in the three proteoglycans were different, and the location of that in glycosaminoglycan chains was also thought to be different. It was demonstrated that three types of proteoglycan which are structurally different are present in extracellular matrix. PMID- 11913493 TI - Viscoelastic behavior of osteoarthritic cartilage. AB - We have studied the incremental stress-strain behavior of human articular cartilage in tension in an attempt to understand the molecular basis for fibrillation and fissure formation in osteoarthritis. Our results indicate that the elastic spring constant for collagen in the direction per pendicular to the cleavage line pattern is about 1.6 GPa (2.3 GPa after correction for the collagen content) and the collagen fibril length is between 0.558 pm at low strains and 1.24 pm at high strains for normal cartilage. Values for the elastic spring constant and collagen fibril length were both found to decrease in OA. The value of the elastic spring constant for collagen perpendicular to the cleavage line pattern is similar to that calculated based on stress-strain curves reported by Kempson. Our results indicate that the elastic spring constant for collagen and the collagen fibril length decrease as the extent of fibrillation and fissure formation increase. Decreases in the elastic spring constant of collagen are consistent with loss of the superficial layer, degradation of proteoglycans and collagen, and subsequent mechanical fatigue. However, changes in the polymer volume fraction are consistent with enzymatic degradation preceding mechanical disruption. It is concluded that osteoarthritic changes to cartilage involve enzymatic degradation of matrix components and fibril fragmentation that is promoted by subsequent mechanical loading. PMID- 11913494 TI - Detection of Mycobacterium tuberculosis antigens in urinary proteins of tuberculosis patients. AB - A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) that can detect up to 0.5 ng of culture filtrate proteins (CFPs) of Mycobacterium tuberculosis strain H37Ra is described. This detection system features several special characteristics: (i) the use of CFPs from the H37Ra strain of Mycobacterium tuberculosis to generate capture and detection antibodies in rabbits and mice, respectively; (ii) affinity purification of CFP-reactive antibodies and selection of the antibody preparations for best performance in the sandwich ELISA system; and (iii) the use of urine-derived protein preparations for antigen detection. The sandwich ELISA could detect up to 0.5 ng of CFPs of the H37Ra strain of Mycobacterium tuberculosis. The assay did not detect antigens of Escherichia coli, Candida albicans or Saccharomyces cerivisiae but efficiently detected CFP preparations from nine different clinical isolates of Mycobacterium tuberculosis. Significant variations, however, were noted in the relative efficacy of the assay to detect CFPs from different clinical isolates of Mycobacterium tuberculosis. The procedure was utilized for detecting tubercular antigens in urine samples from 29 patients with confirmed (sputum-positive) tuberculosis and from 25 healthy controls. Significant levels of antigen could be detected in 22 of the 29 samples tested. PMID- 11913495 TI - Treatment of toxoplasmosis in pregnancy: concentrations of spiramycin and neospiramycin in maternal serum and amniotic fluid. AB - Toxoplasma infection during pregnancy is widely treated with oral spiramycin to reduce the risk of congenital toxoplasmosis in the infant. Failures of therapy have been observed, however. In this study, a sensitive high-performance liquid chromatography technique was used to measure concentrations of spiramycin and neospiramycin, one of the major metabolites of spiramycin, in maternal serum and amniotic fluid. Samples were obtained from 18 women who underwent amniocentesis for polymerase chain reaction (PCR) diagnosis of fetal infection 5-109 days following the prescription of spiramycin therapy (3 g/day). Concentrations of spiramycin and neospiramycin in both serum and amniotic fluid were highly variable, ranging from nondetectable values to 1 microg/ml. None of the concentrations measured were within the range reported to inhibit growth of the parasite in vitro. Consistent with previous reports, part of the observed variability in maternal and fetal drug concentrations could be explained by individual differences in several pharmacokinetic parameters: intestinal absorption, tissue distribution, cellular uptake, metabolism, transfer across the placenta, drug accumulation in fetal tissue, and maternal and fetal drug elimination. The heterogeneity of the data could also be related to differences in patient compliance with the medication prescribed. By addressing factors that could impair adequate treatment of toxoplasmosis during pregnancy, the data presented call for a larger-scale controlled study to determine individual and diurnal variations in maternal drug levels, patient compliance, and outcomes of the offspring. The activity of neospiramycin against Toxoplasma gondii should be assessed. PMID- 11913496 TI - Investigation of a slaughterhouse-related outbreak of Q fever in the French Alps. AB - The aim of the study presented here was to describe the different epidemiological methods used to investigate an outbreak of Q fever that occurred in the spring of 1996 among inhabitants of Briancon, a small town in the French Alps. Three approaches were used: (i) a comparison between a 2-month exhaustive serological survey among blood donors and a retrospective serological survey performed on frozen plasma collected by the transfusion centre in the spring of 1995; (ii) a serological survey performed in the general population by cluster sampling, using dried blood on blotting paper; and (iii) a case-control study. A total of 29 cases of acute Q fever were diagnosed by physicians during hospitalisations of the patients or ambulatory care. The case-con-trol study suggested that the outbreak resulted from airborne transmission of contaminated sheep waste, which had been left uncovered in the slaughterhouse area. Such transmission may have been facilitated by the nearby heliport. The comparison between the cumulative incidence of Q fever among blood donors during the spring seasons of 1995 and 1996 confirmed the outbreak (0.38% vs. 2.58%, respectively; P<0.0001). Health authorities promptly decided to close the slaughterhouse. The use of complementary epidemiological methods allows investigators to focus on major issues related to an outbreak: timely detection of cases, identification of the source, estimations of incidence, and public health intervention. Rapid recognition and management of outbreaks in the general population of a rural region need to be improved, particularly at a time when airborne agents could be used as biological weapons. PMID- 11913497 TI - Quantitative analysis of hepatitis B virus DNA by real-lime amplification. AB - Diagnostic assays allowing the quantification of hepatitis B virus (HBV) DNA over a wide range of concentrations are important for monitoring patients during antiviral therapy. The aim of this study was to develop a new real-time method for HBV DNA quantification. Primers and probe were selected in a highly conserved region of the HBV S gene, and a plasmid containing the pre-S/S region was used as a standard. Linear quantification of the standard was obtained between 10 and 10(9) copies/reaction, with high correlation between ayw and adw genomes (P<0.001). HBV DNA was detected in serial dilutions of a high-titer serum sample with linear results until 2.4 x 10(3) copies/ml. One hundred eight serum samples positive for hepatitis B surface antigen were tested in both the real-time assay and the Digene Hybrid Capture assay (Digene, USA). HBV DNA could be detected by both assays in 70 samples, with significant correlation of results (P<0.001). Results for 38 samples were below the sensitivity limit of the Digene assay, but they could be quantified by the real time polymerase chain reaction assay. These results show that real-time polymerase chain reaction allows sensitive, rapid and linear quantification of HBV DNA in serum. PMID- 11913498 TI - Comparison of the bactericidal activities and post-antibiotic effects of the Des F(6)-quinolone BMS-284756, levofloxacin, and ciprofloxacin against methicillin susceptible and methicillin-resistant Staphylococcus aureus. AB - The bactericidal activities and post-antibiotic effects of BMS-284756 (T-3811ME), levofloxacin, and ciprofloxacin were evaluated against a methicillin-susceptible and a methicillin-resistant Staphylococcus aureus strain. Minimum inhibitory concentrations (MICs), minimum bactericidal concentrations, post-antibiotic effects, and post-antibiotic sub-MIC effects were determined and time-kill studies were performed for BMS-284756, levofloxacin, and ciprofloxacin. At 4 times and 10-times the MIC, time-kill kinetics over 3 h and over 24 h were similar for all three quinolones when effects were considered as multiples of the MIC. All three quinolones achieved a 3 log10 reduction in cfu/ml within 2 h. At 10-times the MIC, the post-antibiotic effects of BMS-284756, levofloxacin, and ciprofloxacin were 1.6-2.6 h for the methicillin-susceptible Staphylococcus aureus strain and 1.5-1.9 h for the methicillin-resistant Staphylococcus aureus strain. When actual concentrations were considered, BMS-284756 achieved results comparable to levofloxacin and ciprofloxacin at concentrations nearly 10-fold less. When relating the pharmacokinetic properties of the three quinolones to their in vitro activities, the resulting Cmax/MIC and AUC/MIC ratios were. respectively, 120-240.7 and 1,321.7-2,643 for BMS-284756, 22.8 and 190 for levofloxacin, and 5.9-11.9 and 54.8-109.6 for ciprofloxacin. The greater in vitro activity and favorable human pharmacokinetics of BMS-284756 may translate to improved clinical effectiveness of this agent compared to currently marketed quinolones. PMID- 11913499 TI - Risk factors for penicillin resistance and mortality in Korean adults with Streptococcus pneumoniae bacteremia. AB - A retrospective analysis was performed to measure the incidence of pneumococcal bacteremia and to identify risk factors for penicillin resistance and prognostic factors for outcome in adults. A total of 151 cases of pneumococcal bacteremia were identified from 149 adults during the period 1996-2000. The overall rate of penicillin resistance was 49%, ranging from 54.2% in 1996 to 48.5% in 2000 (P=0.93). Rates of resistance to ceftriaxone, clindamycin, erythromycin, and trimethoprim-sulfamethoxazole were 21.6%, 51%, 62%, and 44.7%, respectively. Multidrug resistance was documented in 47.7% of the cases. Penicillin resistance was significantly associated with solid tumor, biliary drainage catheter, and previous beta-lactam therapy in the univariate analysis. However, the associations were not as significant as independent risk factors in the multivariate analysis. Mortality was 23.8% and did not change significantly during the study period (P=0.06). Mortality rates in cases caused by penicillin susceptible Streptococcus pneumoniae and penicillin-resistant Streptococcus pneumoniae were 23% and 24.7%, respectively (P=0.81). Mortality was not significantly influenced by penicillin resistance, even high-level resistance (24.4% vs. 20%; P=0.64). Multivariate analysis revealed that antineoplastic chemotherapy, respiratory failure, and acute renal failure were independent prognostic factors for mortality. In conclusion, the rate of penicillin resistance among pneumococcal blood isolates was high in the late 1990s, but penicillin resistance, and even high-level penicillin resistance, was not significantly associated with increased mortality in adults with pneumococcal bacteremia. PMID- 11913500 TI - Three cases of bacterial meningitis after spinal and epidural anesthesia. AB - During a 3-year period, three cases of bacterial meningitis developing after spinal or epidural anesthesia were observed at one hospital in Germany. The causative organisms were Streptococcus salivarius (2 cases) and Staphylococcus aureus (1 case). In the first two cases, contamination of the needle by oropharyngeal flora of the anesthesiological team was likely but remained unproven. In the third case, a nasal swab obtained from the operating anesthesiologist yielded a Staphylococcus aureus strain whose genotypic profile was identical to that of the patient's strain. Infection control procedures for spinal anesthesia are discussed. PMID- 11913501 TI - Epidemiological and clinical features of leptospirosis in Israel. AB - The epidemiology and clinical features of 46 cases of human leptospirosis diagnosed in Israel between 1986 and 1999 were analysed. The median patient age was 37.5 years (range, 16-85 years), and the male/female ratio was 43/3. The most common serogroup found was Leptospira icterohaemorrhagiae. The disease was associated with jaundice in 71% of cases, acute renal failure in 62%, rhabdomyolysis in 52%, pancytopenia in 28%, respiratory failure in 14% and disseminated intravascular coagulation in 5%. Leptospirosis occurs sporadically throughout the year, peaking during the summer months. A shift occurred from predominantly agriculture-related serogroups in the 1970s to urban-related serogroups during the study period reported, with Leptospira icterohaemorrhagiae being the dominant serogroup. PMID- 11913502 TI - Xylitol concentrations in the saliva of children after chewing xylitol gum or consuming a xylitol mixture. AB - Xylitol prevents otitis media when given to children regularly five times per day. To find a more convenient dosing schedule, an enzymatic assay was used to measure xylitol concentrations in the saliva of 65 children after giving them xylitol chewing gum or syrup in doses equal to those used in clinical trials. Although concentrations high enough to have an antimicrobial effect were attained, the xylitol disappeared from the saliva within 15 min. This finding indicates that high peak concentrations are more important for efficacy than the amount of time the xylitol concentration exceeds that needed to produce an antimicrobial effect. A schedule with the same single doses given less frequently could be clinically effective in preventing otitis media. PMID- 11913504 TI - Serological and molecular evidence of human granulocytic ehrlichiosis focus in the Bialowieza Primeval Forest (Puszcza Bialowieska), northeastern Poland. AB - Human granulocytic ehrlichiosis (HGE) is an emerging tickborne zoonosis. First described in the USA, it is being increasingly reported from several European countries. This study was undertaken to provide serological and molecular evidence of the occurrence of the HGE focus in the Bialowieza Primeval Forest, located in northeastern Poland. To this end, the seroprevalence of HGE in this area, where Lyme borreliosis and tickborne encephalitis are highly endemic, was determined by means of an indirect immunofluorescence antibody assay. In addition, the frequency of granulocytic Ehrlichia spp. infection in Ixodes ricinus ticks from the same area was estimated using a polymerase chain reaction method with EHR 521 and EHR 747 primers, which amplified a fragment of 16S rDNA. The rate of seropositivity for HGE was 6.2% (8/130 subjects). Individuals seropositive for Lyme borreliosis were more likely to have anti-HGE antibodies than seronegative ones (P<0.05; OR=6.34, 95%CI=1.12-36.98). There was no association between self-reported frequency of tick bites or forestry employment and HGE seropositivity. Sixty of 376 (16%) Ixodes ricinus ticks tested were positive for the Ehrlichia phagocytophila genogroup by polymerase chain reaction. Ehrlichial DNA was present in 59 of 302 (19.5%) adult ticks and in 1 of 74 nymphs (1.4%). There was a significantly higher infection rate among female ticks (32.9%; 49/149) than among male ticks (6.5%; 10/153) (P<0.05). Dual infection with Ehrlichia spp. and Borrelia burgdorferi sensu lato was detected in 10 samples that were positive for ehrlichiae. The results obtained confirm the perpetuation of the HGE agent in the primeval forest ecosystem of northeastern Poland. PMID- 11913505 TI - Chlamydia pneumoniae DNA in peripheral venous blood samples from patients with carotid artery stenosis. AB - In the study presented here, peripheral blood specimens obtained from patients with atherosclerosis were examined for the presence of Chlamydia pneumoniae to determine whether these specimens can be used for routine testing. Chlamydia pneumoniae DNA was detected in 7 of 56 patients with carotid stenosis and in three of four patients with other atherosclerotic diseases, but it was not detected in any of 50 healthy controls or in any of 59 age- and gender-matched patients suffering from other nonatherosclerotic diseases. IgG antibodies indicative of an active Chlamydia pneumoniae infection were detected by microimmunofluorescence in two of nine PCR-positive patients but in none of 41 PCR-negative patients. Four of nine serum samples obtained from PCR-positive patients contained IgA antibodies compared to 5 of 41 samples obtained from PCR negative patients. PMID- 11913503 TI - Evaluation of a single-step serological assay for laboratory diagnosis of Helicobacter pylori infection. AB - A rapid, single-step, in-laboratory qualitative test for the detection of IgG antibodies to Helicobacter pylori in serum (TestPack Plus; Abbott Laboratories, Germany) was evaluated. This test may be used as an alternative to enzyme immunoassays (EIAs). Of 153 adult patients, 110 were defined as Helicobacter pylori positive and 43 as Helicobacter pylori negative by the gold standard, a combination of three tests. The performance characteristics of the TestPack Plus, i.e. sensitivity, specificity, and positive and negative predictive values, were not significantly different from the corresponding values obtained with an EIA used for comparative purposes, the Pyloriset EIA-G test (Orion Diagnostica, Finland). The high positive predictive value (93%) of the TestPack Plus single step serological test makes it a valuable tool for rapid in-laboratory screening purposes, especially in countries with a high prevalence of Helicobacter pylori infection. PMID- 11913506 TI - Splenic infarction associated with acute Brucella mellitensis infection. PMID- 11913507 TI - Ethnic differences in invasive Haemophilus influenzae disease in the West Midlands region of England, UK. PMID- 11913508 TI - Use of universal PCR on cerebrospinal fluid to diagnose bacterial meningitis in culture-negative patients. PMID- 11913509 TI - Comparison of two methods for antifungal susceptibility testing of Trichophyton rubrum. PMID- 11913510 TI - E test susceptibility testing of nosocomial Clostridium difficile isolates against metronidazole, vancomycin, fusidic acid and the novel agents moxifloxacin, gatifloxacin, and linezolid. PMID- 11913511 TI - Thriving: a life span theory. AB - There is a need for aging theories to become holistic and multidisciplinary with a life span focus. A theory is the construction of explicit explanations in accounting for empirical findings. A good gerontological theory integrates knowledge, tells how and why phenomena are related, leads to prediction, and provides process and understanding. In addition, a good theory must be holistic and take into account all that impacts on a person throughout a lifetime of aging. Based on these criteria, the authors created the Theory of Thriving, with a holistic life span perspective for studying people in their environments as they age. This article proposes a theory for studying people over time in a holistic, encompassing manner. PMID- 11913512 TI - Environmental transformations: an integrative review. PMID- 11913514 TI - The North Carolina Eden Coalition: facilitating environmental transformation. AB - The Eden Alternative, created by Harvard educated physician Dr. William Thomas, is a model for transforming skilled care facilities from institutions based on a medical model of care into human habitats that promote human growth. This change is accomplished through decentralizing the organizational structure of the facility to empower front-line care-delivery staff, and through introduction of plants, animals, gardening, and children into the daily lives of residents. Because the Eden Alternative is a trademarked name, only facilities recognized by the Eden Alternative registry are allowed to use the name. However, any facility may adopt the philosophies embodied by the Eden Alternative, and implement the principles for transforming their facility. In 1996, the North Carolina Division of Facility Services formed the North Carolina Eden Coalition (originally named the North Carolina Eden Alternative Coalition). This was the first organization of its kind in the country. The success of the Coalition has led it to become a model for implementation in other states. Currently, North Carolina leads the country in the number of registered Eden Alternative facilities, and has a large number of other facilities that have instituted some form of environmental enhancement. The Coalition functions as a consumer-driven advisory and educational body rather than a formal regulatory standard body, and endorses implementation of the principles and philosophies embodied in the Eden Alternative. The Coalition has proved to be a successful, cooperative effort among regulators, providers, advocates, and other experts to implement the philosophies of the Eden Alternative without regulatory basis. Because the Coalition was formed under the auspices of the Division of Facility Services and operates with its ongoing cooperation, it is in an excellent position to clarify and influence policies and regulatory issues concerning transformation of long term care environments in North Carolina. This article explores the history, structure, function, mission, and goals of the North Carolina Eden Coalition. Examples of issues the Coalition has addressed in promoting environmental transformation in North Carolina's skilled care facilities are also included. PMID- 11913513 TI - Characteristics of nursing homes adopting environmental transformations. PMID- 11913516 TI - Nursing home environments. PMID- 11913515 TI - Snoezelen: a multisensory environmental intervention. AB - Snoezelen is a multisensory intervention delivered in a specially designed room with high-tech instruments. It is especially useful for end-stage patients with Alzheimer's disease. Snoezelen provides an enabling atmosphere in a failure-free environment. It has been a popular intervention in Great Britain and is just beginning to appear in the United States. PMID- 11913517 TI - Empathy--my geriatric experience. PMID- 11913518 TI - Promoting thriving in nursing homes: the Eden Alternative. PMID- 11913519 TI - Nonviral approaches satisfying various requirements for effective in vivo gene therapy. AB - Development of an efficient method of gene introduction to target cells is the key issue in treating genetic and acquired diseases by in vivo gene therapy. Although various nonviral approaches have been developed, any method needs to be optimized in terms of the target disease and transgene product. The most important information required is (i) target cell-specificity of gene transfer, (ii) efficiency, (iii) duration of transgene expression, and (iv) the number of transfected cells following in vivo application of a vector. These characteristics are determined by the properties of the vector used, as well as the route of its administration, biodistribution, interaction with biological components and the nature of the target cells. Cell-specific gene transfer can be achieved by controlling the tissue disposition of plasmid DNA (pDNA), although the interaction of the pDNA complex with biological components might limit the specificity. Various approaches have been reported to increase the efficiency of transgene expression, from cationic lipids/polymers to physical stimuli, but some of those are ineffective under in vivo conditions. The duration of transgene expression is a complex function involving variables including the cell type, transfection method, and plasmid construct. Immune response often reduces the level and duration of transgene expression. In addition, the number of transfected cells is important, especially in cases in which the therapeutic protein localizes within the target cells. Successful clinical application of nonviral gene delivery methods rely on the development of such methods optimized for a particular target disease. PMID- 11913521 TI - Molecular pharmacology of the Na+-dependent transport of acidic amino acids in the mammalian central nervous system. AB - The Na+-dependent transport of L-glutamate (GluT) has been identified in brain tissue more than thirty years ago. Neurochemical studies, performed in various experimental models during 1970's, defined the basic rules for the selection or synthesis of GluT-specific substrates and inhibitors. The protein molecules (transporters) that mediate the translocation of the substrates across the plasma membrane have been cloned and studied during the last ten years. The sites on the transporters that bind the substrates favour glutamate-like or aspartate-like molecules with one positively charged and two negatively charged ionised groups. Substituents at C3 and C4 are often tolerated but substitutions at C2 or alterations of the ionisable groups usually impede the binding. The substrate binding sites display an "anomalous" selectivity towards stereoisomers. These structural requirements are shared by all Na+-dependent glutamate transporters thus making the design of transporter-selective ligands a challenging task. Moreover, the molecular mechanisms of the transport have not yet been adequately elucidated. Data from a wide variety of experimental studies strongly indicate that Na+-dependent GluT regulates the functioning of the glutamatergic excitatory synapses-the most important rapid inter-neuronal signalling system in the mammalian brain. Altered structural and/or functional properties of the Na+ dependent glutamate transporters have been implicated in the damage to the brain tissue following cerebral ischaemia and in the progressive loss of neurons in conditions such as Alzheimer dementia and amyotrophic lateral sclerosis. Furthermore, it seems that fine-tuning of glutamatergic neurotransmission by regulating the Na+-dependent GluT could be useful in the therapy of schizophrenia. PMID- 11913520 TI - Antiparkinsonian drugs and their neuroprotective effects. AB - In Parkinson's disease, while dopamine (DA) replacement therapy, such as with L DOPA (levodopa), improves the symptoms, it does not inhibit the degeneration of DA neurons in the substantia nigra. Numerous studies have suggested that both endogenous and environmental neurotoxins and oxidative stress may participate in this disease, but the detailed mechanisms are still unclear. Recent genetic studies in familial Parkinson's disease and parkinsonism have shown several gene mutations. This new information regarding its pathogenesis offers novel prospects for effective strategies involving the neuroprotection of vulnerable DA neurons. This review summarizes current findings regarding the pathogenesis and antiparkinsonian drugs, and discusses their possibilities of targets to develop novel neuroprotective drugs. PMID- 11913522 TI - Purification and characterization of mouse mevalonate pyrophosphate decarboxylase. AB - Mevalonate pyrophosphate decarboxylase (MPD) in mouse liver was purified by affinity chromatography. The purified enzyme was a homodimer of 46-kDa subunits and had an isoelectric point of 5.0. Kinetic analysis revealed an apparent Km value of 10 microm for mevalonate pyrophosphate. The enzyme required ATP as a phosphate acceptor and Mg as a divalent cation, which could be substituted with Mn or Co. Its optimum pH was 4.0-7.0. A comparison with MPD from various other sources revealed the mouse MPD to have essentially the same properties as rat MPD, expect for the optimum pH range. An excess of rabbit anti-rat MPD antibody deleted approximately 80% of the MPD activity in the crude extract of mouse liver. These results suggested that the homodimer of 46-kDa subunits represents the major active form of MPD in mice. PMID- 11913523 TI - Enhancement of cellular adenosine triphosphate levels in PC12 cells by extracellular adenosine. AB - To elucidate the biological significance of extracellular adenine compounds, the effects of adenosine (Ado) on cellular levels of adenine compounds, especially adenosine triphosphate (ATP), in PC12 cells were studied. Ado and inosine but not adenosine 5'-monophosphate, adenosine 5'-diphosphate, ATP, guanosine, cytosine, thymidine, and uridine, significantly enhanced cellular ATP levels in PC12 cells in time- and dose-dependent manners. Various P1 receptor agonists of Ado did not enhance the ATP level. In addition, theophylline, an antagonist of P1 receptors, did not inhibit the Ado-evoked ATP enhancement. These results suggest that the Ado receptor is not involved in the augmentation of the cellular ATP level induced by Ado in PC12 cells. The ATP-enhancing effect of Ado was potentiated by dipyridamole, an inhibitor of Ado uptake, or coformycin, an inhibitor of Ado deaminase. The effect of Ado on the ATP level was also observed when PC12 cells were incubated in glucose-free medium. Together these results suggest that enhancement of cellular ATP levels in PC12 cells by extracellular Ado might be acceleration of ATP synthesis through the Ado salvage system using hypoxanthine guanine phosphoribosyltransferase rather than Ado kinase since 5'-iodotubercidin, an inhibitor of Ado kinase, had no effect on the enhancement elicited by Ado. PMID- 11913525 TI - Effect of antiallergic drugs on interleukin 5-induced eosinophil infiltration of rat airways. AB - Interleukin (IL)-5 is thought to play important roles in asthma and to be a potential therapeutic target. An intratracheal injection of murine recombinant IL 5 (3-30 microg/animal) induced a dose-dependent increase in the number of eosinophils in the bronchoalveolar lavage fluid of Brown Norway (BN) rats 24 h after administration. Bovine serum albumin (30pg/animal), used as reference material, did not cause any change. The reaction was not observed in F344 rats. The increase in the number of eosinophils did not accompany bronchial hyperreactivity in BN or F344 rats. Prednisolone (3-10 mg/kg, i.p.) and emedastine (30 mg/kg, p.o.) reduced the increased number of eosinophils induced by the IL-5 challenge. These results suggest that IL-5 is a potent inducer of eosinophils in the airway of BN rats. Prednisolone and emedastine are effective against IL-5-induced eosinophilia. PMID- 11913524 TI - Characterization of synthetic lung surfactant activity against proinflammatory cytokines in human monocytes. AB - Our previous study demonstrated that the smallest synthetic peptide with the sequence CPVHLKRLLLLLLLLLLLLLLLL, SP-CL16(6-28), admixed with phospholipid (synthetic lung surfactant, SLS) showed strong surface activity. In this study, we attempted to develop a dual-type surfactant with both anti inflammatory and surface activities. SP-CL16(6-28) was first chemically synthesized and then purified for use by centrifugal partition chromatography. A mixture of SP-CL16(6 28) and phospholipid complex was tested for anti inflammatory activity using the human monocyte cell line THP-1. Whether the suppression of tumor necrosis factor alpha (TNF-a), interleukin (IL)-8, IL-6, IL-1beta, and macrophage migration inhibitory factor (MIF) was reduced by lipopolysaccharide (LPS) in monocytes was examined. Levels of these cytokines were measured by enzyme-linked immunosorbent assay. It was found that SLS significantly and dose dependently inhibited the secretion of TNF-alpha by THP-1 cells following stimulation with LPS. Dipalmitoylphosphatidylcoline did not inhibit the release of cytokines. These findings suggest that SLS has anti inflammatory activity. Therefore it should be possible to develop a SLS with both anti inflammatory activity and surface activity. PMID- 11913526 TI - Effect of manganese on guinea pig ventricle: initial depression and late augmentation of contractile force. AB - Effects of Mn2+ on isolated guinea pig ventricular myocardia were examined. In isolated papillary muscles, Mn2+ produced a transient decrease in contractile force followed by a late sustained augmentation. Mn2+ markedly increased the amplitude of post-rest contractions; the time course of potentiation was almost the same as that of the late augmentation of contractile force after Mn2+ application. Mn2+ also increased the amplitude of rapid-cooling contractures. The negative inotropic effect of diltiazem and nicardipine was not affected by the presence of Mn2+. Mn2+ shortened the action potential duration under normal condition whereas it prolonged the duration under Ca2+ free conditions. Mn2+, when applied to fura-2-loaded ventricular myocytes, markedly quenched the cytoplasmic fluorescence excited at 360 nm wavelength. We concluded that Mn2+ not only causes a decrease in contractile force by blocking the L-type Ca2+ channel, but also enters the cytoplasm through the channel and produces late augmentation of the contractile force through enhancement of sarcoplasmic reticulum function. PMID- 11913527 TI - Hange-shashin-to raises levels of somatostatin, motilin, and gastrin in the plasma of healthy subjects. AB - Hange-shashin-to has been used for chronic hypofunction of the gastrointestinal tract and to improve functional abnormalities of the upper and lower gastrointestinal system. To determine whether the pharmacological effects of Hange-shashin-to are due to gut-regulatory peptide levels, we developed a sensitive and specific double-antibody enzyme immunoassay (EIA) for detecting motilin and also examined the levels of somatostatin-, motilin-, gastrin-, and vasoactive intestinal peptide (VIP)-immunoreactive substances (IS) in plasma from healthy subjects. We developed a sensitive (3.5 pg, 1.4 pg/well) and specific (carboxy-terminal region) EIA for motilin. A single oral administration of Hange shashin-to 6.0 g caused significant increases somatostatin-IS (20-60 min), motilin-IS (40 min), and gastrin-IS (40-90 min) levels in plasma compared with levels in a placebo group. Hange-shashin-to had no significant effect on VIP-IS levels after single administration. These changes in hormone levels (somatostatin, motilin, and gastrin) might relate to normalization of the upper and lower gastrointestinal system by Hange-shashin-to. PMID- 11913528 TI - DNA toxicity of cocaine hydrochloride and cocaine freebase by means of DNA image analysis on Tetrahymena pyriformis. AB - An assay for computerized scoring of the DNA content of the protozoan Tetrahymena pyriformis has been used for the detection of toxic responses to cocaine, since DNA is responsible for the replication of the genetic material and also reflects closely the number of chromosomes in the nucleus. Thus, doubling of the number of chromosomes in a cell will also correspond to doubling of the DNA content and of the nuclear volume. Two chemical forms of cocaine were used, cocaine hydrochloride and cocaine freebase (crack), at two doses of 1 and 2 mg per 100 ml of protozoan culture, respectively. Image analysis of the protozoan nucleus patterns revealed a rapid stimulating effect on the DNA content for both cocaine hydrochloride and freebase after 1 h of incubation. However, after 2 h of treatment a reduction, although not statistically significant, of the DNA content of the protozoan was observed. These observations were further correlated with the phagocytic activity of the protozoan cultures. This paper provides some possible explanations of the toxic effects of cocaine on this particular cell model. PMID- 11913529 TI - Pharmacological profile of 6,12-dihydro-3-methoxy-1-benzopyrano[3,4-b] [1,4]benzothiazin-6-one, a novel human estrogen receptor agonist. AB - Pharmacological studies were carried out to characterize further the endocrinological profile and the binding mode to the estrogen receptor (ER) of 6,12-dihydro-3-methoxy-1-benzopyrano[3,4-b][1,4]benzothiazin-6-one (1). Binding experiments were conducted with highly purified recombinant human estrogen receptors hERa and beta. Potent estrogenic activity of compound 1 was assessed by testing its ability to down-regulate ERs and to enhance estrogen receptor element (ERE)-dependent transcription. The latest step of our work dealt with the synthesis of the 9-fluorinated derivative 15 for ionic microscopy experiments to determine the intracellular localization of compound 1. Although 1 failed to compete with [3H]E2 for binding to both ER isoforms, evidence was reported that it interacted with hERalpha in MCF-7 cells (ER down-regulation/ERE-dependent luciferase induction). Hence, an appropriate conformation of the hormone binding domain, most probably conferred by co-regulators of ER, is required for the onset of an activity of the compound 1. Estrogenic activity was weak but on the order of magnitude of that of coumestrol (slightly weaker). The synthesis of the 9 methoxylated derivative 16 and its pharmacological evaluation led us to propose a binding mode of 1 on hERalpha. Compound 1 appears to interact with ERa mainly through interactions of its 3-methoxy substituent with the residue His-524 of the hormone binding domain. PMID- 11913530 TI - Prediction of plasma concentration of GTS-21 in hairless rats following monolithic transdermal delivery. AB - The transdermal therapeutic systems (TTS) usually achieve constant plasma concentration for an extended period of time. This is because a sufficient drug stored in the device can keep the constant concentration on the surface of the stratum corneum during the system application. When the drug molecules are not enough to provide the constant surface concentration, the rate of drug penetration decreases with time because of decreased supply of the drug molecules from the delivery device. This paper has proposed an empirical simple approach to predict the plasma concentration for such a TTS. A novel compound, GTS-21, for Alzheimers' disease currently under development was used as a model drug. In vivo and in vitro experiments were carried out in hairless rats. The in vivo plasma concentration-time profile in hairless rats following the application of TTS well agreed with the predicted profile based on the skin pharmacokinetic model together with the model parameters determined from the in vitro experiment. PMID- 11913531 TI - Evaluation of myopathy risk for HMG-CoA reductase inhibitors by urethane infusion method. AB - Purpose of the present study was to evaluate the myopathy risk using a urethane infusion method following oral administration of five kinds of commercial HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) reductase inhibitors (HCRIs), (pravastatin (PV), simvastatin (SV), cerivastatin (CeV), atorvastatin (AV), and fluvastatin (FV)) alone or with coadministration of bezafibrate (BF). The solubility of HCRIs in various solvents was determined as a criterion of the physicochemical property. The plasma creatine phosphokinase (CPK) level as a marker of myopathy in normal rats was screened under urethane infusion after oral administration of HCRI alone or with BF coadministration. Also, renal tissue specimens were prepared and the myoglobin remaining in the tissue was visualized by the labeled avidin-biotin technique. The plasma CPK level in normal rats under urethane infusion following oral administration of five kinds of HCRI increased as the dose of HCRI increased, and coadministration of BF further increased the CPK level for each drug. The risk of myopathy evaluated from the CPK level was ranked as follows: CeV>FV>AV>SV>PV. Myoglobin deposition was observed in the cast of proximal tubules, cytoplasm of distal tubules and collecting ducts of rat kidney extracted from rats treated with HCRIs under urethane infusion. Histopathological findings showed that the extent of myoglobin deposition increased on coadministration of BF with each drug. The correlation was found for myopathy risk evaluated by CPK level using the urethane infusion method and drug lipophilicity, ie., the water/n-octanol partition coefficient except for the case of SV. Histopathological findings for the kidney following HCRI treatment also reflected the CPK level in rats under urethane infusion. PMID- 11913532 TI - NADPH-oxidase may contribute to IL-12 production in macrophages stimulated with CpG phosphorothioate oligodeoxynucleotides. AB - CpG oligodeoxynucleotides (CpG-ODN) could be expectes to act as an adjuvant for humoral and cellular immunities. The aim of this study was to clarify the mechanism of macrophage activation induced by CpG-ODN. Macrophage activation by CpG-ODN was evaluated by measurement of IL-12 secretion. IL-12 secretion by Cp ODN was dose-dependent and saturation was observed at 1 microM. A reactive oxygen species (ROS) scavenger, N-acetylcysteine, inhibited IL-12 secretion, suggesting that the intracellular redox state is important for the activation of macrophages by CpG-ODN. IL-12 secretion was also inhibited by NADPH-oxidase inhibitors, suggesting that ROS generated by NADPH-oxidase is involved in the immunostimulatory effect of CpG-ODN. Nuclear factor (NF)-kappaB is one of the transcriptional factors involved in IL-12 gene expression, and indeed CpG-ODN activated NF-kappaB. However, NADPH-oxidase inhibitors did not affect the activation. These results indicated that NADPH-oxidase plays a crucial role in macrophage activation by CpG-ODN and NF-kappaB does not act as a predominant transcription factor for IL-12 secretion. PMID- 11913533 TI - Uptake of FH by two types of scavenger-like receptors in rat liver parenchymal cells in primary culture. AB - Several anionic proteins that are known to be substrates of scavenger receptors documented in the literature were selected and tested for their effects on the uptake of fractionated heparin (FH), an anionic macromolecular drug. The tests were made in rat liver parenchymal cells to characterize scavenger-like receptors involved in FH uptake, probing into substrate recognition characteristics in comparison with those of scavenger receptors. Although the uptake of FH was completely inhibited by dextran sulfate, a typical substrate of scavenger receptors, suggesting that scavenger-like receptors that have affinity for some anionic macromolecules are responsible for the uptake, it was not inhibited by acetylated low density lipoprotein (Ac-LDL), another typical substrate. Thus, the scavenger-like receptors were suggested to be different from the major scavenger receptors of classes A and B that are known to be sensitive to Ac-LDL. The uptake of FH was only partially inhibited by maleylated bovine serum albumin (Mal-BSA), suggesting that the scavenger-like receptors can be classified into two types in terms of sensitivity to Mal-BSA. The Mal-BSA-sensitive receptor was also found to be sensitive to oxidized low density lipoprotein (Ox-LDL). Kinetic analysis revealed that the binding capacity (Bmax) of the Mal-BSA-insensitive receptor was significantly larger than that of the Mal-BSA-sensitive one, though their dissociation constants (Kd) and apparent internalization rate constants (kint,app) were comparable. Information obtained in this study should be helpful for understanding the disposition mechanism of FH and also of anionic macromolecules and for developing delivery strategies, although the physiological roles and molecular identity of each receptor need to be further clarified in the future. PMID- 11913534 TI - Prediction of human skin permeability using a combination of molecular orbital calculations and artificial neural network. AB - This study was carried out to develop a novel method for predicting the skin permeability coefficient (log K(p)) of compounds from their three-dimensional molecular structure using a combination of molecular orbital (MO) calculation and artificial neural network. Human skin permeability data on 92 structurally diverse compounds were analyzed. The molecular descriptors of each compound, such as the dipole moment, polarizability, sum of charges of nitrogen and oxygen atoms (sum(N,O)), and sum of charges of hydrogen atoms bonding to nitrogen or oxygen atoms (sum(H)) were obtained from MO calculations. The correlation between these molecular descriptors and log K(p) was examined using feed-forward back propagation neural networks. To improve the generalization capability of a neural network, the network was trained with input patterns given 5% random noise. The neural network model with a configuration of 4-4-1 for input, hidden, and output layers was much superior to the conventional multiple linear regression model in terms of root mean square (RMS) errors (0.528 vs. 0.930). A "leave-one-out" cross validation revealed that the neural network model could predict skin permeability with a reasonable accuracy (predictive RMS error of 0.669). PMID- 11913535 TI - In vitro and in vivo effects of macrophage-stimulatory polysaccharide from leaves of Perilla frutescens var. crispa. AB - The crude polysaccharide (PFB-1) was isolated from the leaves of Perilla frutescens var. crispa by the sequential procedures with hot-water extraction, methanol reflux, and ethanol precipitation. It was further purified by anion column chromatography in order to obtain the partially purified polysaccharide (PFB-1-0). In the presence of PFB-1-0, strong cellular lysosomal enzyme activity of murine peritoneal macrophages was observed in vitro. Compared to bacterial lipopolysaccharide (LPS), its activity was relatively high. The in vitro phagocytic activity was enhanced by PFB-1-0 as the similar pattern in both gram negative bacteria, E. coli, and gram-positive bacteria, S. aureus with a time dependent manner. We also investigated the production of several mediators by murine peritoneal macrophages upon stimulation with PFB-1 (in vivo) or PFB-1-0 (in vitro). The levels of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha were increased in the presence of PFB-1-0 in vitro. The PFB-1 stimulated the production of interleukin (IL)-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in vivo. Results suggest that the polysaccharide from P. frutescens var. crispa represents an immunopotentiator and biological response modifiers in vitro and in vivo levels. PMID- 11913536 TI - Mass spectrometric separation and determination of N1,N12-diacetylspermine in the urine of cancer patients. AB - An ionspray ionization mass-spectrometric method for the determination of N1,N12 diacetylspermine (Ac2Spm) was developed using 15N-labeled Ac2Spm as the internal standard. Concentrations of Ac2Spm in the urine obtained from 17 cancer patients measured by the present method correlated well with those measured by ELISA, showing the usefulness of the two methods. PMID- 11913537 TI - Effect of sucrose on formation of the beta-amyloid fibrils and D-aspartic acids in Abeta 1-42. AB - Beta-amyloid peptide 1-42 is a major peptide constituent of beta-amyloid fibrils. We investigated the role of sucrose on the deposition and the D-aspartic acid formation in an amyloidogenic peptide 1-42 under physiological conditions. From analyses using thioflavine-T fluorometric assay and electronmicroscopic spectroscopy after 60 h incubation at 37 degrees C, it was found that sucrose retarded the fibril formation in the amyloidogenic peptide. The retardation of the formation of amyloid fibrils by sucrose was suggested to be not due to viscosity but due to disturbance of the assemlby of alpha-helix containing peptides. Moreover, we showed that the formation of D-aspartyl residue, which is found in beta-amyloid fibrils from Alzheimer disease brains, in the amyloidogenic peptide was also retarded in the presence of sucrose. PMID- 11913538 TI - Lafutidine changes levels of somatostatin, calcitonin gene-related peptide, and secretin in human plasma. AB - We examined the effects of the histamine H2-receptor antagonist, lafutidine, on the levels of gastrointestinal peptides (somatostatin, calcitonin gene-related peptide (CGRP), gastrin, secretin, and motilin) in plasma from healthy subjects. After a single oral administration of lafutidine (10 mg), the plasma lafutidine level (186 +/- 13.4ng/ml) was highest in the 60-min sample after administration and then the plasma level fell. Lafutidine caused significant increase in plasma somatostatin levels at 20 to 120 min and in CGRP levels at 40 to 120 min, compared with a placebo group. The physiological release of plasma secretin was reduced by administration of lafutidine, but the medicine did not alter the level of gastrin or motilin. These results suggest that the pharmacological effects of lafutidine on regulation of gastrointestinal functions closely relate to changes of somatostatin-, CGRP- and secretin-immunoreactive substance levels in human plasma. PMID- 11913539 TI - Cytochrome P450 (CYP) expression in human myeloblastic and lymphoid cell lines. AB - To explore the physiological roles of cytochrome P450 (CYP) in peripheral blood cells, we examined which isoforms of CYP families were expressed in human myeloid leukemia cell lines (U937, HL-60 and K562) and lymphoid cell lines (BALL-1, MOLT 4 and Jurkat) by RT-PCR. We observed relatively high expression of CYP1A1, CYP1B1, CYP2A6, CYP2A7, CYP2D6, and CYP2E1 in all cell types, but CYP2A13 and CYP2C9 expression was not detected. Expressions of aryl hydrocarbon (Ah) receptor and Ah receptor nuclear translocator (ARNT), which mediate induction of the CYP1 family, were also detected in all cell types. Cell-type specific expression of CYP3A4 and CYP3A5 was observed in MOLT-4 and K562 cells. Weak, but significant, expression of CYP3A7 was detected in K562 cells. The profile of CYP expression in the culture cells reported here provides information that furthers our understanding of the physiological roles of CYP enzymes in human blood cells. PMID- 11913540 TI - Latent activity of curcumin against leishmaniasis in vitro. AB - In this study the anti-proliferative effect of curcumin (curcuma longa) that is the active ingredient of ground dried rhizome has been studied against three local and three reference leishmanial strains, Leishmania major, Leishmania tropica and Leishmania infantum (Pakistani isolate). Curcumin has shown an average IC50 of 5.3 microM against promastigotes of various leishmanial strains which is much lower as compared with pentamidine that is one of the basic treatments against leishmaniasis. The main draw back attributed to these assays performed on promastigotes is the heterogeneity of results compared with those obtained with intracellular amastigotes or with in vivo effect. We also tested activity of curcumin against axenic amastigote like cells (AALC) of L. major strain (MHOM/PK/88/DESTO). Curcumin proves to be far more potent then pentamidine against AALC which further strengthens the fact about its leishmaniacidal activity. PMID- 11913541 TI - Zeaxanthin dipalmitate from Lycium chinense fruit reduces experimentally induced hepatic fibrosis in rats. AB - We previously reported that zeaxanthin dipalmitate (ZD), a carotenoid from Lycium chinense fruit, reduces myofibroblast-like cell proliferation and collagen synthesis in vitro. To determine whether ZD might reduce the severity of hepatic fibrosis in an animal model, hepatic fibrosis was induced in rats by bile duct ligation/scission (BDL) for a period of 6 weeks. Treatment of BDL rats with ZD at a dose of 25 mg/kg body weight significantly reduced the activities of aspartate transaminase (p<0.05) and alkaline phosphatase (p<0.001) in serum. Furthermore, collagen deposition was significantly reduced as assessed by the Sirius Red binding assay in BDL rats administered ZD at the dose of 25 mg/kg body weight (p<0.01). In addition, the levels of thiobarbituric acid-reactive substances and 4-hydroxyproline were reduced when BDL rats received ZD at the dose of 25 mg/kg body weight. These results showed that ZD effectively inhibited hepatic fibrosis in BDL rats, at least in part via its antioxidative activity. PMID- 11913542 TI - Secretory transport of methylprednisolone possibly mediated by P-glycoprotein in Caco-2 cells. AB - We recently reported that P-glycoprotein (MDR1) is capable of interfering with the absorption of methylprednisolone in the rat small intestine. This study was undertaken to examine the interaction between methylprednisolone and MDR1 using Caco-2 cells. The permeation of various steroid hormones (hydrocortisone, prednisolone, progesterone, beta-estradiol, and testosterone) was compared. The basolateral-to-apical (secretory) permeation of methylprednisolone was more than 3-fold greater than the apical-to-basolateral (absorptive) permeation. When verapamil (0.1 mm), a potent modulator of MDR1, was added to both apical and basolateral sides of Caco-2 cells, the absorptive permeation of methylprednisolone was increased and its secretory permeation was decreased. As a result, the secretory-oriented manner of methylprednisolone permeation almost completely disappeared. Prednisolone and hydrocortisone exhibited weaker secretory-oriented movement than did methylprednisolone. The secretory-oriented permeation of prednisolone and hydrocortisone was also diminished by the addition of verapamil. There was no significant directionality in progesterone permeation and the permeation of beta-estradiol and testosterone tended to be absorptive. These results appear to suggest that methylprednisolone, prednisolone, and hydrocortisone interact with MDR1 as the substrates. In contrast, there was no evidence that MDR1 was capable of potently interfering with the absorption of the sex hormones tested in this study, supporting our previous findings in the rat. It was further found that apically-added verapamil demonstrated a modulating effect on MDR1 function even at 5 microM. PMID- 11913543 TI - Cytochrome P450 1A1/2 mediated metabolism of trans-stilbene in rats and humans. AB - It was demonstrated that trans-stilbene was metabolically activated to the estrogenic compound by rat liver microsomes (Sugihara et al., Toxicol. Appl. Pharmacol., 167, 46-54 (2000)). In this study, determination of the isoforms of cytochrome P450 involved in the oxidation of the proestrogen, trans-stilbene, to its hydroxylated metabolites was examined. When trans-stilbene was incubated with rat liver microsomes in the presence of NADPH, estrogenic compounds, trans4 hydroxystilbene and trans-4,4'-dihydroxystilbene were formed. Comparison of the oxidase activity among liver microsomes of untreated, 3-methylcholanthrene treated, acetone-treated, clofibrate-treated, dexamethasone-treated and phenobarbital-treated rats toward trans-stilbene showed that those from 3 methylcholanthrene-treated rats exhibited the highest activity. Human liver microsomes also catalyzed the oxidation in varying degrees. Variation in trans stilbene oxidase activity was closely correlated to that of phenacetin O deethylase activity. The oxidase activity was inhibited by alpha-naphthoflavone; however, in this case trans-4,4'-dihydroxystilbene was not detected. The oxidase activity toward trans-stilbene was exhibited by recombinant human cytochrome P450 1A1 and 1A2 expressed in a human B lymphoblastoid cell line. PMID- 11913544 TI - Effects of epidermal growth factor (EGF) on fetal islet B cells in vitro. AB - ABSTRACT. It has been reported that when the rat fetus is treated with streptozotocin (STZ) in vivo, islet B cells are destroyed but later recover. To investigate the process of the recovery of B cells after in vitro treatment of the fetal pancreas with STZ and the role of epidermal growth factor (EGF) in the recovery of B cells, we measured the level of insulin released from the cultured fetal pancreas and examined it histologically. As a result, we immunohistologically confirmed the regeneration of B cells in the pancreas that had been cultured for 48 hr after destruction of islet B cells by STZ treatment. An immunohistologic study using proliferating cell nuclear antigen (PCNA) showed that without the addition of EGF, the cell division index was significantly higher in the STZ-treated group (STZ group) than in the untreated group (intact group), whereas with the addition of EGF, the cell division index increased in both groups, but EGF did not have a significant cell division-promoting effect on the pancreas in the STZ group. The addition of EGF caused a significant decrease in the concentration of insulin in culture medium in both groups. These results indicate that EGF has a cell growth-promoting effect on intact fetal pancreas in vitro but has the effect of inhibiting the release of insulin, and thus suggest that EGF does not trigger the regeneration of islet B cells. PMID- 11913545 TI - Administration of estradiol-3-benzoate down-regulates the expression of testicular steroidogenic enzyme genes for testosterone production in the adult rat. AB - ABSTRACT. To study the role of estrogen in the testes, testosterone and testicular steroidogenic enzyme mRNA levels were investigated in male Sprague Dawley rats 24 hr after intramuscular administration of a single dose of estradiol-3-benzoate (EB). EB administration resulted in a greater decrease in intra-testicular and serum testosterone in 10-week-old rats than in 3- or 5-week old rats. A dose of 2 microg EB/kg had the lowest observed effect. The level of serum luteinizing hormone (LH) was unchanged at any dose. Semiquantitative RT-PCR analysis revealed that, of the four major testicular steroidogenic enzymes, mRNA levels of cytochrome P450 side-chain cleavage and 17beta-hydroxysteroid dehydrogenase type-III were significantly reduced, and mRNA levels of cytochrome P450 17alpha-hydroxylase/ C17-20 lyase (P450c17) were reduced severely and significantly, by EB administration. However, the level of 3beta-hydroxysteroid dehydrogenase type-I mRNA was not changed. In addition, the P450c17 mRNA level in EB-treated rats was much lower than that in the testes of hypophysectomized rats, with the level in the latter being equal to that in control rats. LH is secreted into blood periodically, the effects of estrogen on the LH secretion pattern of the pituitary gland, for example, in frequency and amplitude of LH pulse, were difficult to detect with the methods of the present study. The results indicated, at least, that EB administration down-regulates P450c17 gene expression predominantly, resulting in the inhibition of testosterone production. From the differences in the steroidogenic enzyme expressions between hypophysectomized and EB-treated rats, it was suggested that EB acts on the testis directly or indirectly though not via alteration of LH secretion and induces reduction of P450c17 mRNA level. PMID- 11913547 TI - Effects of LH and PGF2alpha in equine dominant follicles observed by MDS. AB - Microdialysis System (MDS) is a novel technique used for investigation of molecule secretion between different cell populations. Local hormonal secretion at follicular wall has been still unclear. This MDS study was used to determine progesterone (P4), androstenedione (A4), estradiol-17beta (E2) and Prostaglandin F2alpha (PGF2alpha) release in mare pre-ovulatory follicles. Follicles larger than 30 mm were isolated from the ovary and follicular fluid aspirated for hormone assay. Follicular fluid collected from small, middle and large follicles were analyzed by EIA. The concentrations of P4 and PGF2alpha were similar among the different sizes of follicles. The release of A4 was observed in middle and large follicles. E2 concentration was observed in middle follicles and was higher in large follicles compared with middle follicles. Follicular wall was cut and incubated for MDS and when LH was infused, there was an increase in P4 and A4 release. PGF2alpha release was considerably high after LH infusion compared to the control group. Infusion of PGF2alpha increased P4 and A4 release but there was no change in E2 release. This results suggest that in pre-ovulatory follicles, LH stimulates theca interna cells and also PGF2alpha seemed to have a mediator role to induce steroid hormone production and luteinization of follicular cells. The nature of the mechanisms involved in selection of large follicles is still a perplexing research problem in reproduction. PMID- 11913546 TI - Detection and identification of the Candida species by 25S ribosomal DNA analysis in the urine of candidal cystitis. AB - Candida species in clinical urine samples were identified directly by the newly developed method of PCR analysis on 25S ribosomal DNA (rDNA). Two dogs were referred to the Animal Medical Center, Nihon University School of Veterinary Medicine, Fujisawa, Kanagawa, Japan for the examination of chronic cystitis. Microscopic examination of urine samples from these dogs revealed yeast cells. Urine culture on Sabouraud's dextrose agar at 27 degrees C for 5 days produced white to cream colored colonies. The isolates were identifical to Candida albicans and C. parapsilosis by mycological examination, respectively. The nucleotide sequences of 25S ribosomal DNA from these urine isolates showed 99% similarity to those of a reference strain of Candida albicans or C. parapsilosis. The nucleotide sequences of 25S rDNA obtained directly from urine samples were also identical to C. albicans and C. parapsilosis, respectively. Confirming the results on the isolates cultured from the same urine samples. This PCR analysis method could be available for the direct identification of Candida species in urine samples within 2 days. PMID- 11913548 TI - Expression of desmosomal proteins in rat keratinocytes during in vitro differentiation. AB - The keratinocyte, the major component of the epidermis, expresses several proteins that characterize the keratinization during the differentiation. Proliferation and differentiation of cultured human keratinocytes are known to be regulated by the Ca2+ concentration in the culture medium. However, informations about the rat keratinocyte are relatively limited and their physiology is still an open question. To elucidate the characteristics of the rat keratinocyte, we established rat keratinocyte culture system and examined effects of extracellular calcium concentration on the expression of differentiation-related proteins. Keratinocytes were isolated from the newborn rat skin with 0.25% trypsin, followed by separation with a Percoll density gradient. The separated cells were grown in MCDB 153 medium containing several growth factors and Ca(2+)-free fetal bovine serum, then stimulated with Ca2+. Immunoblotting demonstrated strong expression of beta1 integrin in unstimulated cells, suggesting that the primary culture of rat keratinocytes was successfully established. Expression of desmoglein and transglutaminase was increased by Ca2+ stimulation, whereas beta1 integrin expression was decreased in response to increasing concentrations of Ca2+. These observations indicate that cultured rat keratinocytes maintain the ability to differentiate in vitro, which is similar to that of the basal keratinocytes in the epidermis. PMID- 11913550 TI - Cranial features of the spotted seal, Phoca largha, in the Nemuro Strait, considering age effects. AB - Cranial features (development, individual variation, and sexual dimorphism) were examined from the 23 metrical characters and 2 nonmetrical characters (the degree of closure of the 9 cranial sutures and the presence of sagittal crest) in the two spotted seal specimen groups at the Nemuro Strait, Hokkaido. One specimen group was incidentally taken in the salmon trap nets between 1982 and 1983 (n = 70), and the other was randomly sampled by damage control kill between 1997 and 1998 (n = 82). The development of morphometrical characters of skulls ceased at 5.6, 10.7, 7.9, and 11.9 yr. old, for 1982-83 male, 1982-83 female, 1997-98 male and 1997-98 female, respectively. The sutures were half ankylosed till approximately 10 yr. old in both sexes. The sagittal crest began from about 5 yr. old in male. Individual variation of skull was large in the feeding, breathing, and facial-expression apparatus. On the other hand, the variation of braincase, and skull concerning to the movement of head/neck tended to be small. Only 1997 98 specimens exhibited a sexual dimorphism in skull characters except for the braincase, whereas the dimorphism was not found in 1982-83 specimens. We could not detect the significant difference between two specimen groups, although there were a few differences in characters related to the rostrum and mandible. PMID- 11913549 TI - Serous cysts are a benign component of the cyclic ovary in the guinea pig with an incidence dependent upon inhibin bioactivity. AB - Ovaries were collected from normal cycling female guinea pigs on each day of the estrous cycle (n = 5 per day) for histological analysis of ovarian morphology. Three types of ovarian cysts were observed: serous cysts, follicular cysts and parovarian cysts. The most common were serous cysts (cystic rete ovarii), which were present throughout the estrous cycle with an overall incidence of 63.5% (54 out of 85 animals). Follicular cysts occurred in 22.4% of guinea pigs overall (19 out of 85). Only one parovarian cyst (1 out of 85) was observed in the present experiment. Follicular cysts always coincided with serous cysts and were less common during diestrus. The incidence of serous cysts did not vary significantly across the estrous cycle. In a second experiment, cycling female guinea pigs were arrested in a prolonged luteal phase by a progesterone implant in order to achieve ovarian synchrony. They were then treated with inhibin antiserum (0.5 or 1 ml per animal i.v.; n = 6 per group) or normal goat serum (controls; n = 6 per group). There was a dose dependent increase in the incidence of serous ovarian cysts following passive immunization against the inhibin alpha-subunit. These results suggest that serous cysts are a normal component of the cyclic guinea pig ovary and that alterations in the inhibin-follicle-stimulating hormone system appear to modulate the incidence of serous ovarian cysts in this species. PMID- 11913551 TI - Detection of Babesia microti-like parasite in filter paper-absorbed blood of wild rodents. AB - The first case of human babesiosis was reported in Japan. The epidemiology of this disease in Japanese nature remains unclear. In this study, 97 common field mice captured in Hokkaido, Japan, were examined. Blood specimens absorbed onto filter papers were eluted and tested by nested PCR using specific primers for the B. microti nuclear small subunit rRNA genome. Twenty-three percent (11/47) of Apodemus speciosus and four percent (2/50) of Clethrionomys rufocanus were positive. The 159-bp primary sequences of PCR products tested exhibited 97.5% and 96.8% homology with those of the human isolate in Japan and of U.S. strains of B. microti, respectively. PMID- 11913552 TI - Gastroduodenal adenocarcinomas and rectal adenoma in a cougar (Felis concolor) infected with Helicobacter-like organisms and spirochetes. AB - A 14-year-old female cougar died from gastroduodenal adenocarcinomas and rectal adenoma. At necropsy, polypoid tumor masses of various sizes were scattered on the mucosal surfaces of the stomach, duodenum, and rectum. Histologically, the gastric tumor was diagnosed as an intestinal type adenocarcinoma and the tumor cells metastasized to the mesenteric lymph nodes, spleen, and lung. Helicobacter like organisms were detected in the lumina lined by foveolar epithelium. In the duodenum, the carcinoma cells were localized in the limina propria and many of them were intensely positive for proliferating cell nuclear antigen (PCNA). In contrast, the rectal adenoma had a lower number of PCNA-positive cells. In the rectum, chronic inflammation with numerous spirochetes was also noted. These results indicated that the occurrence of the gastrointestinal tumors might be associated with the bacterial infection described above. PMID- 11913553 TI - Cerebellar myxoid type meningioma in a Shih Tzu dog. AB - A 6-year and 9 month-old, male, Shih Tzu dog showed ataxia and trembling. By MRI examination, a mass (1 cm) was found in the right cerebellum. As the dog did not respond to radiation therapy, and showed a rise of intracranial pressure, he was euthanized. The cerebellar mass was soft and hemorrhagic. Histologically, the mass contained vimentin-positive spindle- or polyhedral-shaped cells arranged in a cord-like pattern. Mucinous materials were observed in the intercellular spaces. Ultrastructural examination revealed cell processes, microtubule-like structures and desmosomes. The case was diagnosed as myxoid type meningioma. PMID- 11913554 TI - Isolation of Salmonella from diarrheic feces of pigs. AB - A survey of Salmonella was carried out in fecal samples of 887 pigs with diarrhea collected from 235 pig farms between April 1996 and March 2001. Salmonella was isolated from 84 feces (9.5%) of 887 pigs and from 45 (19.1%) of 235 farms. The higher prevalence was found in weaned pigs (12.4%) and fattening pigs (17.3%) than in sows (4.2%) and suckling pigs (4.5%). Isolation rates of S. Typhimurium were higher from weaned and fattening pigs than from the others. Therefore, risk of horizontal infection of S. Typhimurium will increase, if no adequate health managements are practiced when weaned and fattening pigs have diarrhea. PMID- 11913555 TI - Studies on serological cross-reaction of Neospora caninum with Toxoplasma gondii and Hammondia heydorni. AB - The enzyme-linked immunosorbent assay (ELISA) was used to examine cross reactivity of Neospora caninum with Toxoplasma gondii and Hammondia heydorni. Anti-T. gondii mouse and cat sera cross-reacted with N. caninum soluble antigen (NLA), but not with the recombinant surface antigen (NcSRS2). Anti-H. heydorni dog sera showed no cross-reactivity with either the NLA antigen or the NcSRS2. Lack of cross-reactivity between anti-H. heydorni sera and N. caninum antigens, and the cross-reactivity of anti-T. gondii sera with the NLA suggest that N. caninum has common antigens to T. gondii except for NcSRS2 based on serology. In light of several studies suggesting a closer relationship between N. caninum and H. heydorni than with T gondii, examination of serological cross-reactivity with N. caninum may be necessary to further classify the parasites in addition to molecular and morphological studies and clarification of the life cycle. PMID- 11913556 TI - Identification of genotypes of Cryptosporidium parvum isolates from a patient and a dog in Japan. AB - Cryptosporidium parvum (C. parvum) is recognized as a significant pathogen in humans and animals, primarily as a cause of diarrheal illness. Recent genetic and biological studies indicate that C. parvum is not a single species but composed of genetically distinct multiple genotypes. Thus, it is valuable to distinguish between genotypes in the epidemiology of Cryptosporidium infection in humans and animals. Although C. parvum has been detected in humans and animals in Japan, the genotype of isolates remains unclear because identification has been performed only by conventional microscopy. We report herein the genotypes of C. parvum isolates distinguished by the polymerase chain reaction (PCR)-based diagnostic method. C. parvum isolates, originally obtained from a patient and a pet dog, were found to have cattle and dog genotypes, respectively. PMID- 11913557 TI - Effect of natural and synthetic retinoids on the proliferation and differentiation of three canine melanoma cell lines. AB - The effect of two natural retinoids and synthetic retinoids with or without retinoid synergists on the proliferation and differentiation of 3 melanoma cell lines were investigated in vitro. No retinoids showed significant growth inhibitory effect on these cell lines when used alone, however, cell differentiation and significant growth inhibition were observed when treated with a combination of retinoids and a retinoid synergist. This study may suggest that, though the cells showed low susceptibilities when retinoids were treated alone, the combination of retinoids and a retinoid synergist may be effective to control the growth of canine melanoma cell lines. PMID- 11913558 TI - Hyaluronidase is not essential for the lethality of Erysipelothrix rhusiopathiae infection in mice. AB - To investigate the role of hyaluronidase in the pathogenicity of Erysipelothrix rhusiopathiae, transposon Tn916 was transferred from Enterococcus faecalis CG110 to a virulent strain of E. rhusiopathiae, and hyaluronidase-deficient mutants were isolated. A virulence assay in the mice showed that of the seven hyaluronidase-deficient mutants tested, six mutants were avirulent, but that one mutant, designated AST121, was as virulent as its parental strain. Western immunoblotting with a monoclonal antibody specific to the capsule, a major virulence factor of the organism, revealed that all of the avirulent mutants had lost the capsular antigen, whereas the mutant AST121 did not. These results suggest that the lack of virulence of the six hyaluronidase-negative mutants could be due to a loss of the capsule and that hyaluronidase does not contribute to the lethality of E. rhusiopathiae infection in mice. PMID- 11913559 TI - The increment of anti-ORP150 autoantibody in initial stages of atheroma in high fat diet fed mice. AB - Expression of 150 kda oxygen-regulated protein, ORP150, was examined in the atheromatous lesions on aortic valves in high-fat diet fed mice. Immunohistochemical staining revealed that ORP150 was expressed on the surface of plaque and was co-localized with phagocytes bearing Mac-3, a mouse macrophage differentiation antigen. These findings suggest that ORP150 is involved in the development of the atheromatous plaque. Titer of autoantibody against ORP150 was gradually elevated in parallel with the length of period of high-fat diet feeding. These results suggest that the deposition of immunocomplex toward ORP150 antigen is involved in atheromatous plaque progression. PMID- 11913560 TI - High level expression of C-terminal truncated recombinant chicken interferon gamma in baculovirus vector system. AB - Chicken interferon-gamma (ChIFN-gamma) was expressed by baculovirus in a C terminal truncated form, namely ChIFN-gammaT, to accelerate the secretion of the expressed protein. It is also expressed as ChIFN-gammaT bearing poly His tag, ChIFN-gammaTHis, for easy purification. The expressed proteins were detected by SDS-PAGE analysis with Coomassie brilliant blue staining. The purified ChIFN gammaTHis with nickel chelated column showed anti-viral activity in vitro and stimulation of the secretion of nitrogen intermediates such as nitric oxide in chicken peripheral blood mononuclear cells. Antiserum against ChIFN-gammaTHis recognized the 15 kDa, 16 kDa, and 32 kDa bands that seemed to be an unglycosylated monomer, a glycosylated monomer, and a homodimer of ChIFN gammaTHis in the culture supernatant, respectively. The anti-serum also recognized around 14 kDa and 28 kDa bands in the sera of chickens or concanavalin A stimulated spleen cell culture supernatants that seemed to be monomeric and dimeric forms of a natural ChIFN-gamma, respectively. PMID- 11913561 TI - The cerebellum: it's about time! But timing is not everything--new insights into the role of the cerebellum in timing motor and cognitive tasks. AB - Converging evidence from different research studies supports a role for the cerebellum in timing neural processes. The cerebellum is part of a distributed system for motor control. The timing hypothesis provides a specific functional role for the unique contribution of the cerebellum. The timing capabilities of the cerebellum appear to extend beyond motor control into tasks focusing on perceptual processing that require the precise representation of temporal information and sensorimotor learning. Behavioral and modeling studies suggest that the cerebellar timing system is best characterized as providing a near infinite set of interval-type timers rather than as a single clock with pacemaker or oscillatory properties, but this is controversial. In addition to learning precisely timed motor responses, the cerebellum is involved in on-line processing using feed-forward systems for which sensory input is used prior to movement execution to improve movement accuracy. This would be a mechanism for triggering accurate "time." The cerebellum continues to fascinate scientists, and although survival is possible without the cerebellum, the resultant quality of life is significantly compromised with clumsiness, ataxia, hypotonia, dysarthria, slowing of various cognitive perceptual processes, and impaired fine motor and ocular motor coordination. The last three decades have seen the development of research that has focused on how the cerebellum functions. Further neurophysiologic research in cerebellar cortical neurotransmission is likely to further our understanding of the cerebellar contribution to timing sensorimotor processes. PMID- 11913562 TI - Hemiconvulsion-hemiplegia-epilepsy syndrome: characteristic early magnetic resonance imaging findings. AB - We report three patients with hemiconvulsion-hemiplegia-epilepsy syndrome who presented acutely and were shown to have striking neuroimaging findings suggestive of diffuse cytotoxic edema confined to one hemisphere, including extensive diffusion-weighted imaging abnormalities in two cases. Two patients subsequently developed progressive and extensive atrophy of the involved hemisphere. These findings are consistent with earlier descriptions of the classic neuroradiologic features of this syndrome and are helpful in the differential diagnosis of acute infantile hemiplegia. Further, the findings support the previously proposed pathogenetic mechanism of neuronal injury caused by status epilepticus. PMID- 11913563 TI - Benign childhood epilepsy with centrotemporal spikes: clinical characteristics and identification of patients at risk for multiple seizures. AB - This study aimed to characterize patients diagnosed as having benign childhood epilepsy with centrotemporal spikes and few seizures and compare them with patients with benign childhood epilepsy with centrotemporal spikes with multiple seizures. The medical files of 87 consecutive patients with benign childhood epilepsy with centrotemporal spikes were reviewed and data on gender, age at disease onset, duration of the disease, number of seizures, seizure semiology, and electroencephalographic and neuroimaging findings were analyzed. The mean age at disease onset was 8 years. The mean duration of the disease was 2 years. Eighteen percent of the patients had more than 20 seizures each, whereas 24% had 1 to 3 seizures each. The only predictor for a disease course with multiple seizures was an onset prior to 3 years of age. PMID- 11913564 TI - Rett syndrome: clinical manifestations in males with MECP2 mutations. AB - Rett syndrome is a neurodevelopmental disorder characterized by cognitive and adaptive regression with autistic features, loss of acquired skills, and stereotypic hand movements that almost exclusively affects females. It is an X linked dominant disorder, with presumed lethality in males. Nonetheless, there are a few descriptions of males suspected of having Rett syndrome. With the recent discovery that the MECP2 gene is responsible for most cases of Rett syndrome, it is possible to molecularly assess cases of affected males by direct sequencing analysis. We describe an Israeli family consisting of a female having classic Rett syndrome and a male sibling with severe neonatal encephalopathy. Molecular analysis revealed that both sister and brother have the same MECP2 gene mutation; however, their mother does not. This case, as well as other published studies of males with MECP2 mutations, reveals that the clinical manifestations in viable males vary from neonates with severe encephalopathy to adults with mental retardation and demonstrate genotype-phenotype correlations. PMID- 11913565 TI - Subacute sclerosing panencephalitis: clinical and magnetic resonance imaging evaluation of 36 patients. AB - We studied 36 patients (24 males, 12 females), all of whom had definite subacute sclerosing panencephalitis with typical periodic complexes in their electroencephalograms and increased titers of measles antibody in serum and cerebrospinal fluid. Their clinical and laboratory findings on admission were reviewed retrospectively. The age at onset of symptoms varied from 4 to 23 years. The average age at onset of disease was 13.1 +/- 4.18 years. The mean of the duration from the infection to the onset of subacute sclerosing panencephalitis was 9 years. Unusual symptoms, especially in the early periods of disease, included hemiparesis (7 patients), headache (3), generalized tonic-clonic seizures (6), absence seizure (1), nausea (3), and vomiting (3). Twenty-six cranial magnetic resonance imaging (MRI) and 12 computed tomography examinations were performed. Nine patients had normal MRI. In the early stages, lesions usually involved parieto-occipital corticosubcortical regions asymmetrically. In time, symmetric periventricular white-matter changes became more prominent. In addition to the common clinical findings in cases of subacute sclerosing panencephalitis reported in the literature, there were some different clinical features of the disease. Eventually, we concluded that there seems to be no correlation between the clinical stages and either the duration from the onset of subacute sclerosing panencephalitis or the MRI findings. PMID- 11913566 TI - Presence of the 5,10-methylenetetrahydrofolate reductase C677T mutation in Puerto Rican patients with neural tube defects. AB - Folic acid supplementation can reduce the incidence of neural tube defects. The first reported genetic risk factor for neural tube defects is a C677T mutation in the 5,10-methylenetetrahydrofolate reductase gene, resulting in decreased activity of the enzyme. We examined the enzyme mutation role of methylenetetrahydrofolate reductase in the etiology of neural tube defects in our population. The study group consisted of 204 Puerto Rican individuals including 37 pregnant females with a prenatal diagnosis of neural tube defects in their fetuses, 31 newborns, 36 fathers, and 100 healthy adults. The prevalence of the C677T mutation was examined. Homozygosity for the alanine to valine substitution (TT) was observed in 9% of the controls and 19% of the mothers with children with neural tube defects. Our results indicate that the presence of the T allele at the methylenetetrahydrofolate reductase 677 position may increase the risk of giving birth to an infant with a neural tube defect. PMID- 11913567 TI - Mutation analysis of MECP2 and clinical characterization in Korean patients with Rett syndrome. AB - Rett syndrome is a progressive neurodevelopmental disorder occurring predominantly in females. Recently, mutations in the MECP2 gene on Xq28, which encodes methyl-CpG binding protein 2, were identified as responsible for some cases of Rett syndrome. In the present study, we analyzed the entire coding sequence of the MECP2 gene in 20 sporadic cases of Rett syndrome in Korea. Of the 20 patients, 14 (70%) had pathogenic mutations, which included 10 different mutations. Altogether, there were five missense mutations (D97Y, L100V, R133C, T158M, R306C), four nonsense mutations (R168X, R255X, R270X, R294X), and one frameshift mutation (a 41-bp deletion at 1157-1197). Two of these were novel mutations (D97Y, L100V). Most of the nucleotide substitutions involved C to T transitions at CpG hotspots. We could find no clear phenotype-genotype correlation according to the type of mutation. However, there was a tendency for patients with no MECP2 mutation (30%) to show more severe symptoms and more rapid clinical progression than patients with mutations. Further studies are necessary to identify the other possible genetic causes of Rett syndrome. PMID- 11913568 TI - Relationship of epilepsy-related factors to anxiety and depression scores in epileptic children. AB - Cognitive and behavioral impairments are found more often among epileptic children than among their peers. In this study, we evaluated the anxiety and depression in epileptic children to compare their results with that of a healthy control group and to determine the relationship of anxiety and depression scores to epilepsy-related factors. The State Trait Anxiety Inventory (STAI) and Children's Depression Inventory (CDI) were applied to 35 patients with epilepsy aged 9 to 18 years (mean age 12.9 +/- 2.52 years) and to 35 healthy children who served as the control group. Both study and control groups were divided into two age groups (9 to 11 and 12 to 18 years) to exclude the effect of puberty on anxiety and depression scores. Significant depression and suicidal ideation were determined in the study group. The mean trait anxiety score was significantly higher in the 9- to 11-year age group of epileptic patients than the corresponding control group (35.90 +/- 6.90 and 29.33 +/- 2.84, P < .05). The mean state anxiety score (33.90 +/- 3.90 and 30.40 +/- 6.02, P < .05), trait anxiety score (38.20 +/- 6.84 and 32.20 +/- 3.90, P < .05), and depression score (16.65 +/- 8.32 and 8.15 +/- 3.15, P < .05) were significantly higher in the 12- to 18-year age group of epileptic children than in the control group. Among the epilepsy-related factors, whereas epilepsy duration, seizure frequency, and polytherapy were determined to increase anxiety and depression, age of seizure onset, seizure type, and electroencephalographic findings were not related to anxiety and depression. Symptoms of anxiety and depression are common among epileptic children, especially during puberty. The State Trait Anxiety Inventory and Children's Depression Inventory may be used as a tool to provide information to clinicians. PMID- 11913569 TI - Coagulation abnormalities and acquired von Willebrand's disease type 1 in children receiving valproic acid. AB - Valproic acid is commonly used in the management of childhood epilepsy. The known hematologic side effects of the drug are hemorrhagic diatheses, thrombocytopenia, and hypofibrinogenemia. We studied coagulation parameters in 29 epileptic children receiving valproic acid for at least 6 months. Their ages ranged between 2 and 18 years (10.2 +/- 4.9 years). The total valproic acid dose was 250 to 1000 mg/day equivalent to 20 to 30 mg/kg/day. Treatment duration ranged from 6 to 57 months. These children had not previously had a hemostatic defect and had no family history of bleeding disorders. Platelet count, prothrombin time, activated partial thromboplastin time, bleeding time, fibrinogen, platelet aggregation assays, and ristocetin cofactor activity levels were determined in all of the patients, but von Willebrand's factor antigen levels could be determined in only 14 patients. The values of von Willebrand's factor antigen ranged from 53 to 218% (104.1 +/- 42.3), and ristocetin cofactor activity levels ranged from 11.5 to 218% (94.5 +/- 43.1). Six of the 29 children (20.7%) had decreased values of ristocetin and cofactor activity and were considered to have acquired von Willebrand's disease. The decreases in coagulation parameters were not dependent on either valproic acid dose or treatment duration. Two patients with low ristocetin cofactor activity values had mild epistaxis, which did not require discontinuation of therapy. In patients receiving valproic acid therapy, this side effect must be considered, especially before surgical intervention and serious traumatic conditions. PMID- 11913571 TI - Diffuse magnetic resonance imaging white-matter changes in a 15-year-old boy with mitochondrial encephalomyopathy. AB - We report a 15-year-old boy who presented with recurrent headaches associated with numbness, confusion, and speech difficulty. Brain magnetic resonance imaging (MRI) revealed diffuse bilateral white-matter hyperintensity on fluid-attenuated inversion-recovery and T2-weighted images in the internal capsule, periventricular and subcortical white matter, base of the pons, and middle cerebellar peduncles. Lesions were isointense on T1-weighted images and nonenhancing. Muscle biopsy showed changes consistent with a mitochondrial myopathy. Mitochondrial respiratory chain enzyme activity analysis revealed a significant reduction in complex II activity. Genetic testing was negative. We conclude that mitochondrial encephalomyopathy may present with unusual diffuse supratentorial and infratentorial white-matter changes on MRI. PMID- 11913570 TI - Gelastic seizure with hypothalamic hamartoma: proton magnetic resonance spectrometry and ictal electroencephalographic findings in a 4-year-old girl. AB - Gelastic seizure is a rare symptom often associated with hypothalamic hamartoma. We present here a 4-year-old girl with gelastic epilepsy caused by hypothalamic hamartoma and report the magnetic resonance spectrometry and electroencephalographic (EEG) findings. At the age of 2 1/2 years, she developed brief, repetitive laughing attacks or mixed attacks with laughing and crying, which were refractory to carbamazepine. An interictal EEG showed intermittent slow waves in the left frontocentral region and sporadic positive sharp waves in the left centroparietal area. Ictal EEG demonstrated dysrhythmic theta activity in the left central area 3 seconds after the onset of laughing. Brain magnetic resonance imaging demonstrated a large sessile mass, isointense to gray matter, in the region of the hypothalamus, suggesting hypothalamic hamartoma. Proton magnetic resonance spectrometry of the hypothalamic hamartoma revealed a significant reduction of the N-acetylaspartate/serum creatinine ratio. The altered chemical shift imaging with magnetic resonance spectrometry in our patient suggests a biochemical abnormality in the tissue of the hypothalamic hamartoma. Moreover, this abnormal function of the hamartoma tissue might be closely related to epileptogenesis because the time difference between the ictal laughter and the subsequent EEG changes in the ictal EEG does not support the idea that the activated cortex is the epileptogenic focus. PMID- 11913572 TI - Neurodevelopmental and behavioral abnormalities associated with deletion of chromosome 9p. AB - We report a child with craniosynostosis, partial absence of the corpus callosum, developmental delay, precocious puberty, and deletion of chromosome 9(p12p13,3). A review of the literature did not reveal any previous combination of the same kind. Craniosynostosis and partial absence of the corpus callosum, separately or in conjunction, may be part of the spectrum of malformations in the chromosome 9p deletion syndrome, and its presence, in combination with other known features, should prompt a search for this particular deletion as part of the differential diagnosis. PMID- 11913573 TI - Posterior cerebral artery occlusion associated with Mycoplasma pneumoniae infection. AB - Cerebral infarction is a rare complication of Mycoplasma pneumoniae infection. In all cases previously reported in the literature, vascular occlusion occurred in the anterior brain circulation, either the internal carotid or the middle cerebral artery. We report a case of a child with posterior cerebral artery occlusion and resultant hemiparesis associated with M. pneumoniae infection. PMID- 11913574 TI - Prevalence of hypopigmented maculae and cafe-au-lait spots in idiopathic epileptic and healthy children. AB - The cutaneous lesions and findings related to the central nervous system are frequently seen concomitantly in many patients. Neurocutaneous syndromes are the most typical examples supporting this observation. The prevalences of hypopigmented maculae and cafe-au-lait spots were investigated in 210 idiopathic epileptic children between the ages of 2 and 17 years and 2754 healthy children between the ages of 5 and 15 years. In the group of epileptic children, hypopigmented maculae and cafe-au-lait spots were observed in 30 (14.3%) and 63 (30%) children, respectively. In the group of healthy children, the prevalence of hypopigmented maculae was 1.6% (44 children) and of cafe-au-lait spots was 2.83% (78 children). The difference between the two groups was very significant statistically (P < .0001). PMID- 11913576 TI - Reversible vascular changes in children with cerebral infarction. AB - A case of cerebral infarction in a 4-year-old male is described. The child presented with an acute onset of right hemiplegia, central facial palsy, and dysarthria. He had no predisposing factors for cerebral infarction. A computed tomography scan showed a diffuse low-density area in the territory of the left miiddle cerebral artery. Magnetic resonance angiography disclosed multiple irregular narrowings in the left anterior and middle cerebral arteries. He recovered spontaneously from the stroke with minimal long-term complications, and repeated angiography disclosed a complete regression of the vascular changes 2 months after the stroke. There was no recurrence of stroke after 2-year follow up. This case demonstrates the importance of longitudinal angiographic follow-up in childhood cerebral infarction of idiopathic origin. PMID- 11913575 TI - Hypomagnesemic seizures: case report and presumed pathophysiology. AB - Hypomagnesemia has previously been recognized as an uncommon cause of seizures. The electrolyte abnormality is caused by poor gastrointestinal absorption or excessive renal wasting, both from a variety of causes. We report on a 5-week-old patient who developed hypomagnesemic seizures as a consequence of renal magnesium wasting. Although the exact pathophysiology of hypomagnesemic seizures remains uncertain, currently available information suggests that it is related to disinhibition of the N-methyl-D-aspartate-type glutamate receptor-sodium channel complex. PMID- 11913577 TI - Pontocerebellar hypoplasia in two siblings with dysmorphic features. AB - We present two siblings with pontocerebellar hypoplasia who have progressive microcephaly, mental and motor retardation, truncal ataxia, strabismus, and progressive spasticity and hyperreflexia of the lower limbs. Extrapyramidal dyskinesia and epilepsy, other main clinical features of pontocerebellar hypoplasia, are absent. The older sibling also has a high arched palate, triangular-shaped face, thoracolumbar scoliosis, pectus carinatum, kyphosis, cubitus valgus, arachnodactyly, long extremities, and a tall stature, which were not previously reported in association with pontocerebellar hypoplasia. The clinical phenotype should be expanded, especially within type II, with the reports of additional cases. PMID- 11913578 TI - Role of apoptosis in Duchenne's muscular dystrophy. AB - We investigated the presence of apoptosis in muscle tissues from 24 patients (average age 5.44 +/- 1.81 years) with Duchenne's muscular dystrophy by in situ tailing of nuclear fragmentation. Muscle tissue from 4 children without histologic evidence of myopathy served as normal controls. Muscle fibers positive for nuclear DNA fragmentation were determined quantitatively by counting an area of at least 400 muscle fibers. Eleven of 24 specimens showed no nuclei with DNA fragmentation. On the other hand, 0.37 +/- 0.48% of fibers in patients with Duchenne's muscular dystrophy and none in controls had DNA fragmentation (P > .05). In this study, the percentage of apoptotic nuclei was higher in Duchenne's muscular dystrophy muscle than in normal controls. However, the difference did not reach a statistically significant level, and further studies with larger control groups are warranted. PMID- 11913579 TI - Multiple ring-enhancing lesions in a child with relapsing multiple sclerosis. AB - The presence of ring-enhancing lesions in brain magnetic resonance images (MRIs) often raises the concern of an infectious etiology, although this radiographic finding is also seen in patients with multiple sclerosis. Multiple ring-enhancing lesions have been reported in adult patients diagnosed with multiple sclerosis but have not yet been reported in childhood multiple sclerosis. We report here a 14-year-old girl with recurrent neurologic symptoms. Her initial brain MRI showed multiple ring-enhancing lesions involving numerous white-matter fiber tracts. An extensive investigation for infectious etiologies was unrevealing. Studies of cerebrospinal fluid showed an elevated myelin basic protein and the presence of an oligoclonal band not seen in the serum. The results of electrophysiologic studies suggested a demyelinating process. The patient responded rapidly to high dose corticosteroid treatment. However, she suffered a clinical relapse 3 months later, presenting with dysesthesia and weakness of the right arm. Repeat MRI showed multiple new active lesions. This case report illustrates that multiple ring-enhancing lesions in the brain MRI can be seen in children with multiple sclerosis and that multiple sclerosis should be considered as part of the differential diagnosis when encountering a pediatric patient with similar radiographic findings. PMID- 11913580 TI - Lumbar spinal stenosis causing congenital clubfoot. AB - Congenital lumbar spinal stenosis is believed to rarely cause neurologic symptoms during childhood. We present a 16-year-old boy with bilateral congenital clubfeet surgically corrected by tendo Achillis releases at 2 years of age who presented with progressive, bilateral footdrop. Magnetic resonance imaging of his lumbosacral spine showed severe spinal stenosis at the L3-5 vertebrae. Congenital lumbar spinal stenosis is probably an under-recognized cause of lower extremity neurologic abnormalities, including clubfoot deformity. Magnetic resonance imaging has made this eminently treatable disorder easier to recognize. PMID- 11913581 TI - Lambert-Eaton myasthenic syndrome in children. AB - Lambert-Eaton myasthenic syndrome is a presynaptic disorder of neuromuscular transmission. It is characterized by muscle weakness, hyporeflexia, and autonomic dysfunction. It is most often associated with small cell carcinomas of the lung. Rare cases have been reported in children. We recently encountered two children with Lambert-Eaton myasthenic syndrome associated with antibodies to P/Q-type calcium channel but without evidence of neoplasms. Both patients showed prolonged and significant improvement following cyclosporin treatment. The diagnosis of Lambert-Eaton myasthenic syndrome should be considered in children with progressive weakness and a negative work-up for the usual causes. High-frequency repetitive nerve stimulation and P/Q-type calcium-channel antibodies may confirm the diagnosis. PMID- 11913582 TI - Syndrome of cerebrospinal fluid hypovolemia following lumbar puncture cerebrospinal fluid leak in a patient with idiopathic intracranial hypertension. AB - An 11-year-old girl presented with headache of 3 months' duration. There was bilateral disc edema. The cerebrospinal fluid pressure was 50 cm of water with normal cerebrospinal fluid cytology and biochemistry. She developed severe headache (different and disabling), dizziness, vomiting, and backache on sitting up 6 hours after lumbar puncture, and lying supine relieved all of her symptoms. Intravenous fluids, analgesics, and complete bed rest did not relieve her symptoms over the next 72 hours. She was completely relieved of her symptoms on receiving two tablets of Caffergot containing 200 mg of caffeine and 2 mg of ergotamine 72 hours after lumbar puncture. The symptoms recurred 48 hours later, and a repeat dose of Caffergot was required. Magnetic resonance imaging (MRI) done 96 hours after lumbar puncture revealed the entire dura overlying the brain, including the posterior fossa, showing intense enhancement on contrast injection with leak at the lumbar puncture site. Oral caffeine (coffee, three times a day) was advised over 1 week. The patient remained asymptomatic, and a repeat MRI scan after 10 days showed complete clearing of the cerebrospinal fluid leak with no dural enhancement. The syndrome of cerebrospinal fluid hypovolemia following lumbar puncture is reported in a girl with idiopathic intracranial hypertension. PMID- 11913583 TI - Hemiplegic migraine with prolonged symptoms: case report. AB - Hemiplegic migraine is defined by the occurrence of migraine during attacks of unilateral weakness. Neurologic symptoms last for 15 to 60 minutes in most cases. Attacks usually start in childhood, adolescence, or early adulthood. Diagnosis may be delayed if there is no relevant family history. A 16-year-old girl who was diagnosed with hemiplegic migraine presenting with prolonged left hemiparesis is reported. The importance of this case is that the pediatrician will also consider migraine in the differential diagnosis of a child presenting with hemiparesis even if there is no previous headache and family history. PMID- 11913584 TI - Effect of deposition site and sperm number on the fertility of sheep inseminated with liquid semen. AB - The effect of the deposition site and the numbers of sperm on the fertility of sheep was tested in a field trial in which 1292 Norwegian crossbred ewes aged between six months and five-and-a-half years from 52 farms were inseminated with liquid semen after natural oestrus. Cervical insemination with 150 x 10(6) and 75 x 10(6) spermatozoa resulted in 25-day non-return rates of 63.7 and 56.1 per cent, and vaginal insemination gave non-return rates of 63.3 and 56.6 per cent, respectively. There was no significant difference between the cervical and vaginal inseminations, but the inseminations with 150 x 10(6) spermatozoa gave significantly higher non-return rates (P=0.004). There were significant differences between the non-return rates for different rams (P<0.0001) and farmers (P=0.0002) but the age of the ewe had no significant effect. PMID- 11913585 TI - Myopathy in brown pelicans (Pelicanus occidentalis) associated with rancid feed. AB - Three adult brown pelicans (Pelicanus occidentalis) were observed to be weak, anorexic and unresponsive to antibiotics, anti-inflammatory drugs, vitamins including vitamin E, and steroids. Blood chemistry revealed high activities of aspartate aminotransferase, creatinine kinase and lactate dehydrogenase. Radiographs of the birds' leg muscles revealed multiple opacities suggestive of calcification; the gross lesions included white streaks in the leg, wing, and heart muscles, and the microscopical lesions consisted of various degrees of degeneration and necrosis characterised by eosinophilia, variations in fibre size, loss of striations, myolysis, mineralisation, and proliferation of mononuclear cells in the skeletal muscles and the myocardium. The levels of heavy metals, selenium and vitamin E in the birds' livers were not abnormal. The level of peroxide in their diet of capelin fish was high, 69 meq/kg, (normal <20 meq/kg) consistent with rancid feed, and the level of vitamin E was very low, 0.5 iu/kg (normal 20 to 30 iu/kg). It was concluded that the myopathy was probably caused by vitamin E deficiency due to feeding the pelicans a rancid diet. PMID- 11913586 TI - Reported response to treatment among 245 cases of equine headshaking. PMID- 11913587 TI - Escherichia coli as a cause of mortality in piglets in the royal Kingdom of Bhutan. PMID- 11913588 TI - Serotyping of Swiss avian Chlamydophila psittaci isolates. PMID- 11913589 TI - Effect of Neospora caninum on in vitro development of preimplantation stage bovine embryos and adherence to the zona pellucida. PMID- 11913590 TI - Anthrax outbreak in Mago National Park, southern Ethiopia. PMID- 11913591 TI - Competition Commission's inquiry into the supply of animal POMs. PMID- 11913592 TI - FMD inquiries. PMID- 11913593 TI - PMWS in Argentina. PMID- 11913594 TI - Human disability and the veterinary profession. PMID- 11913595 TI - Copycat or designer original? PMID- 11913596 TI - Support for recent graduates. PMID- 11913599 TI - Extension of the behavioral model of healthcare utilization with ethnically diverse, low-income women. AB - OBJECTIVE: Psychosocial vulnerabilities were added to a model of healthcare utilization. This extension was tested among low-income women with ethnicity addressed as a moderator. DESIGN: Structured interviews were conducted at 2 points in time, approximately 1 year apart. The constructs of psychosocial vulnerability, demographic predisposing, barriers, and illness were measured by multiple indicators to allow use of Structural Equation Modeling to analyze results. The models were tested separately for each ethnic group. SETTING: Community office. PARTICIPANTS: African-American (N = 266), Euro-American (N = 200), and Mexican-American (N = 210) women were recruited from the Dallas Metropolitan area to participate in Project HOW: Health Outcomes of Women, a multi-year, multi-wave study. Face-to-face interviews were conducted with this sample. Participants had been in heterosexual relationships for at least 1 year, were between 20 and 49 years of age, and had incomes less than 200% of the national poverty level. MAIN OUTCOME MEASURES: Healthcare utilization, defined as physician visits and general healthcare visits. RESULTS: Illness mediated the effect of psychosocial vulnerability on healthcare utilization for African Americans and Euro-Americans. The model for Mexican Americans was the most complex. Psychosocial vulnerability on illness was partially mediated by barriers, which also directly affected utilization. CONCLUSIONS: Psychosocial vulnerabilities were significant utilization predictors for healthcare use for all low-income women in this study. The final models for the 2 minority groups, African Americans and Mexican Americans, were quite different. Hence, women of color should not be considered a homogeneous group in comparison to Euro Americans. PMID- 11913600 TI - Sociodemographic differences in exposure to health information. AB - OBJECTIVE: Baseline data from the Heart Attack REACT Study provided the opportunity to explore population subgroup differences in exposure to health information in an ethnically diverse sample from 5 regions across the United States. METHODS: During the 4-month baseline period of the REACT study, some 1,200 residents from the 20 study communities were surveyed using random digit dial telephone methods. Respondents were asked to recall health messages seen and/or heard recently, and the sources of these messages. Comparisons were made between sociodemographic subgroups defined by age, sex, race/ethnicity, education, income, work status, and geographic location. RESULTS: Except for education level differences, the amount of exposure to health information did not vary significantly by sociodemographics; however, significant variation among subgroups in the types of messages cited and the sources of these messages was observed. Minority and low-income groups were found to have less exposure to chronic disease prevention information, eg, on nutrition, exercise, and heart disease. Additionally, the sources of health information most popular among sociodemographic subgroups appeared to be a determining factor in the types of messages received. CONCLUSIONS: The results of these analyses support previous findings, adding to the sparse body of information on the best channels for reaching under-served populations. PMID- 11913598 TI - New salt-sensitivity metrics: variability-adjusted blood pressure change and the urinary sodium-to-creatinine ratio. AB - DESIGN: We report the results of a 24-week, placebo-controlled, two-period, crossover trial of sodium supplementation in 112 normotensive African Americans, aged 25 to 64 years, with average blood pressure (BP) of 105/70 mm Hg. Estimated 24-hour urinary sodium excretion was 133.6 mmol; the average urinary sodium-to creatinine ratio was 0.74. METHODS: Variability-adjusted BP change was the difference in BP level after the respective treatment periods, divided by the intra-person standard deviation of the average BP obtained at 3 consecutive screening visits during a 4-week period. RESULTS: The urinary sodium-to creatinine ratio and the total urine sodium content were 37.8% and 26.5% higher, respectively, at the end of the sodium treatment period. Twenty-four hour ambulatory BP change (mm Hg) (95% CI) was systolic 1.2 (0, 2.4), and diastolic 0.7 (-0.3, 1.8); cuff BP change was systolic 0.9 (-1, 2.9), and diastolic 1.4 (0.1, 2.7). Variability-adjusted BP change was systolic 0.2 (-0.4, 0.8) and diastolic 0.4 (-0.1, 0.9). Though variability-adjusted and unadjusted SBP change correlated highly (r = 0.941, P<.001), only the former correlated with body mass index (r = 0.224, P<.05), a known correlate of salt sensitivity. While total urinary sodium content in timed urine collections and urinary sodium-to creatinine ratio correlated (r = 0.727, P<.001), neither correlated with cuff BP changes. Change in urinary sodium-to-creatinine ratios of 3 consecutive pooled overnight 8-hour urine collections correlated with changes in 24-hour ambulatory SBP (r = 0.294, P<.001) and DBP (r = 0.193, P<.05); however, change in total urinary sodium content was uncorrelated. Total urinary sodium content of these pooled collections (P = .001), but not the urinary sodium-to-creatinine ratio, was positively related to urinary creatinine excretion per kilogram of body weight, the latter being an indicator of urine collection duration. CONCLUSIONS: The lack of effect of the duration of urine collection on the urinary sodium-to creatinine ratio is advantageous in individuals who may report inaccurately the duration of their urine collection. Sequential regression analyses demonstrated that the urinary sodium-to-creatinine ratio conveyed all of the changes in urinary sodium excretion information contained in aggregate urinary sodium excretion-and more. Variability-adjusted BP change was the more sensitive metric of BP response to dietary sodium manipulations, than unadjusted BP change. Thus, variability-adjusted BP change and the urinary sodium-to-creatinine ratio appear to be incrementally better metrics of salt sensitivity than those traditionally used. PMID- 11913601 TI - Categorizing race and ethnicity in the HMO Cancer Research Network. AB - BACKGROUND: The Cancer Research Network (CRN) was formed in 1999 with funding from the National Cancer Institute. The CRN represents a collaboration of 10 health plans across the United States, with a combined total of approximately 9 million enrollees. The goal of the CRN is to promote collaborative research, which will ultimately increase the effectiveness of preventive, curative, and supportive interventions for major cancers. Special emphasis is placed upon diverse populations, and racial and ethnic differences in outcomes, costs, and cost effectiveness. PURPOSE: There is increasing awareness in the research literature of the relationship between race and ethnicity and health outcomes. However, the majority of the health maintenance organizations represented in the CRN, similar to other health plans and organizations, do not routinely collect race and ethnicity data on their members. In order to compare data and outcomes across the CRN sites, consensus is needed in the measurement of race and ethnicity. METHODS: This review discusses terminology used in the research literature to describe race and ethnicity and the manner in which these constructs have been measured in previous studies. CONCLUSIONS: This review concludes with suggestions for standardized measures of race and ethnicity. IMPLICATIONS: It is hoped that shared conceptualizations of race and ethnicity will lead to improved data quality and precision in measurement. PMID- 11913602 TI - The neglected entity of familial cardiac amyloidosis in African Americans. PMID- 11913603 TI - African-American ethnicity in epidemiological studies of calcium antagonists in relation to cancer. PMID- 11913604 TI - The association between racial identity and hypertension in African-American adults: elevated resting and ambulatory blood pressure as outcomes. AB - OBJECTIVES: This study investigated the association between Black racial identity attitudes and hypertension. It was hypothesized that racial identity attitudes characterized, in part, by an intense focus on African Americans as a group and a general rejection of White individuals and culture (termed transitional identity), would be associated with elevated blood pressure. It was also hypothesized that the experience of stress and hostility or cynical mistrust associated with transitional identity would account for this association. METHODS: Participants were 126 non-obese African-American men and women (mean age 53.8 years) with normal blood pressure or minimally treated hypertension, recruited from among individuals enrolled in a study of risk factors for atherosclerosis in southwestern Pennsylvania. Participants completed assessments of racial identity, hostility, perceived stress, and race-focused situational appraisal. Physiological measures included resting and daytime ambulatory systolic blood pressures (SBP) and diastolic blood pressures (DBP), as well as nocturnal declines in blood pressure. RESULTS: Transitional racial identity attitudes significantly predicted resting SBP (P<.03) and DBP (P<.002), as well as ambulatory SBP (P<.001) and DBP (P <.0004), when adjusting for demographic variables. Transitional identity remained a significant predictor of resting DBP (P<.01) and ambulatory SBP (P<.02) and DBP (P<.006), when hostility and perceived stress were also controlled. CONCLUSIONS: Results suggest that transitional racial identity may be an important correlate of elevated blood pressure in African Americans and that this association cannot be fully accounted for by measures of perceived stress or hostility. PMID- 11913605 TI - Differences between Hispanics and non-Hispanic Whites in use of hospital procedures for cerebrovascular disease. AB - OBJECTIVE: This study examined disparities in the use of in-hospital diagnostic and therapeutic procedures for Hispanics with cerebrovascular disease compared to their non-Hispanic White counterparts. DESIGN: This is a cross-sectional study using 1996 hospital administrative data. METHODS: Hispanics and non-Hispanic Whites with diagnosis codes indicating occlusion or stenosis of precerebral arteries or transient cerebral ischemia were included, with a total of 18,674 New York patients (5.1% Hispanic) and 22,624 California patients (11.1% Hispanic). Adjusted odds ratios compared Hispanics with non-Hispanic Whites for six diagnostic and therapeutic procedures for cerebrovascular disease, controlling for patient and hospital characteristics. RESULTS: Hispanics had higher rates of non-invasive diagnostic procedures (head CT scan, head/neck diagnostic ultrasound, echocardiogram, and head MRI). The odds of invasive diagnostic testing (cerebral arteriogram) and therapeutic procedures (carotid endarterectomy) were lower for Hispanics. Most findings remained unchanged in logistic regression models with patient and hospital characteristics. Adding a measure of the concentration of Hispanic patients by hospital eliminated or reduced observed differences between Hispanics and Whites. CONCLUSIONS: Controlling for each hospital's experience with Hispanic patients eliminated or reduced the magnitude of the disparities in procedure use, suggesting that the concentration of Hispanic patients in a hospital is associated with different patterns of procedure use. PMID- 11913606 TI - Eliminating health disparities: the time for action is now. PMID- 11913607 TI - Delayed presentation for care during acute myocardial infarction in a Hispanic population of Los Angeles County. AB - BACKGROUND: Delay in seeking care for acute myocardial infarction (AMI) has been well described in some populations, but little research has been conducted on delay by minority groups, such as Latinos and Asians. This study sought to determine the degree of delay and mode of access of patients with AMI presenting to an inner-city hospital serving an ethnically diverse population. METHODS AND RESULTS: This study was a retrospective case series of Latino, Asian, African American and Caucasian patients diagnosed in the emergency department (ED) with chest pain (CP) and AMI, during a 2-year period. Three hundred thirty seven cases were studied, with the average delay in presentation for care being 44.3 hours, with a median of 8.8 hours. Before seeking care for symptoms of AMI, Latinos delayed an average of 41.5 hours (median 9.2 hours); African Americans delayed an average of 30.8 hours (median 3.5 hours); Asians delayed an average of 92 hours (median 12 hours); and Caucasians delayed an average of 31.6 hours (median 3.2 hours). The mode of transportation used by the different groups to travel to the ED was also significantly different, with Latinos and Asians utilizing private transportation the majority of the time (70% and 83%, respectively). CONCLUSION: There is a significant delay in presentation for care in patients experiencing AMI in Los Angeles County, greatest among the Asian and Latino populations. Once the decision to seek assistance is made, an under-utilization of EMS compounds the problem. Socioeconomic status, language, and cultural practices pose unique barriers to recognizing and addressing symptoms of AMI in this population. PMID- 11913608 TI - Heart disease and its related risk factors in Asian Indians. AB - Although Asian Indians represent the second fastest growing Asian immigrant group in the United States, we know little about their increased risk for coronary artery disease (CAD). A key word search of Medline (using key words Asian Indian, South Asian Indian, coronary artery disease, and heart disease), from 1980-2001, was used to develop a database of articles relating to coronary artery disease for Asian Indians in the United States and abroad. We describe the prevalence and other data of CAD in Asian-Indian communities abroad and in the United States. We then outline certain risk factors for coronary artery disease, specifically diet, cholesterol, and Type 2 diabetes, which contribute to the increased risk of heart disease in Asian Indians. Finally, we describe an approach to screening and potential prevention of coronary artery disease in those of Asian-indian descent in this country. PMID- 11913609 TI - Body image assessment: comparison of figure rating scales among urban Black women. AB - PURPOSE: To determine a psychometrically stable and socially acceptable scale for assessing perceptual body image in Black women. METHODS: The study sample (N = 50) was selected sequentially from Black women (mean age 52.3 +/- 10 years) who were being screened for a larger randomized trial (Project Joy). Three standard published figure rating scales (FRS) were compared with a new scale (Reese FRS). Distributions of respondents' selections on all scales were examined relative to anthropometric measures, including body mass index (BMI) and categories of obesity and overweight (National Expert Panel on Overweight and Obesity Guidelines). Cultural identity, developed from an adaptation from the African American Acculturation Scale II (cross-validation and short form), and its relationship to FRS preference was also assessed. RESULTS: Body weight distribution of women in this study matched that of US Black women over 40 years of age in the NHANES III data. All four FRS performed similarly and correlated significantly with BMI, r was -0.70 to -0.75, P<.0001. Twenty-two percent of the women identified themselves with one of the three largest images (determined by consensus of an expert panel to represent obesity) on three FRS and 38% identified with the three largest images on one scale. For all scales, this was below 56% of women identified as obese based on national guidelines. Among women with the highest cultural identity scores, indicating the strongest identification with elements of Black culture, 72% preferred the new Reese FRS. CONCLUSION: Overall, there is considerable overlap among images selected for any weight category based on national BMI classifications. A strong preference exists for the Reese scale, especially among women with a strong Black cultural identity. This new scale performs similarly to existing scales in successively assessing the self-assignment of an image for self, but is more socially acceptable. Reasons for the small percentage of obese women who identified themselves as obese need further investigation. PMID- 11913610 TI - Arterial elasticity among normotensive subjects and treated and untreated hypertensive subjects: influence of race. AB - OBJECTIVES: To determine arterial elasticity in normotensives and in treated and untreated hypertensive Black and White subjects. METHODS: A prospective multicenter, controlled clinical trial evaluated large (C-) and small (C2) artery elasticity indices among 3 groups: 1) normotensive subjects with or without a family history of hypertension; 2) controlled and treated hypertensive subjects; and 3) untreated and uncontrolled hypertensive subjects. Blood pressure was measured using a mercury manometer and arterial compliance or elasticity was determined using a CVProfilor DO-2020 CardioVascular Profiling System (Hypertension Diagnostics, Inc, Eagan, Minn). These parameters were measured in triplicate 3 minutes apart in a random sequence, with the patient in a supine position. Two-way ANOVA was used for statistical evaluation. RESULTS: One hundred seventy eight subjects met stratification and enrollment criteria. The mean age was 46 years. 109 were White and 69 were Black. [table: see text] CONCLUSION: Small and large arterial elasticity indices are reduced as hypertension status worsens. Age and height were important covariates of C1 and C2. Race was not found to be a significant predictor of either C1 or C2. Large and small artery elasticity indices do not differ between White and Black subjects with varying degrees of hypertension after adjusting for covariates. More studies with a larger number of subjects are required. PMID- 11913611 TI - Disparities in adherence to recommended followup on screening mammography: interaction of sociodemographic factors. AB - OBJECTIVE: The objective of this study was to examine disparities in adherence to screening mammography and, specifically, to investigate whether race/ethnicity, education, age, health insurance, and family history of breast cancer (FHBC), as unique factors and in interactions, influence adherence to recommended follow up on screening mammography. DESIGN: The study involved retrieval and analyses of data collected by the Colorado Mammography Project (CMAP) for 167,232 diverse (82.8% White, 3.4% Black, 11% Hispanic, 1.6% Asian, 0.6% Native American, and 0.6% "other") screening participants during the 1990-1997 study period. METHODS: Subjects' first mammograms captured by CMAP were tracked in the database to identify women who received follow-up recommendations, women who adhered within 12 months and those that did not. Analyses included comparisons of adherence rates among women with various sociodemographic characteristics. RESULTS: Of the 17,358 women who received follow-up recommendations, 80.7% adhered. Overall, non White women in each of the racial/ethnic groups were less likely to adhere to recommendations than were White women (P<.05). Also less likely to adhere were the younger, less educated, uninsured/underinsured, and women who reported not having FHBC. CONCLUSION: Race/ethnicity appeared to interact with age, education, health insurance, and FHBC to influence the probability of adherence, suggesting the need to explore further cultural, psychosocial, and situational factors. PMID- 11913612 TI - Using participant information to develop a tool for the evaluation of community health worker outreach services. AB - OBJECTIVE: Because there is no instrument that measures how clients judge community health worker (CHW) services, we sought to develop such a questionnaire. We report how we used client information to develop a brief questionnaire evaluating CHW services. DESIGN: We conducted and content-analyzed 18 in-depth semi-structured interviews of clients receiving CHW services to determine aspects of care salient to clients. Based on the results of these analyses, we developed and administered an in-person survey measuring the importance of 57 aspects of CHW services to 84 clients in 3 programs using CHWs to help control hypertension or diabetes. RESULTS: Clients perceived a broad array of aspects of CHW care including CHW attributes, services, benefits or outcomes of service and service arrangements. The 15 aspects ranking highest included: 1) CHW knows job; 2) CHW keeps client alive; 3) CHW gives information on high blood pressure; 4) CHW shows respect; 5) blood pressure is lowered; 6) CHW pays attention; 7) client gets better medical care; 8) CHW speaks understandably; and 9) client gets needed care. CONCLUSION: We used client information to generate and determine the relative importance of a pool of aspects that we and others can use to construct brief questionnaires to measure clients' judgments of CHW services. Such questionnaires are needed for ongoing evaluation as more providers and managed care organizations increase their use of CHWs for outreach programs. PMID- 11913613 TI - Is there homogeneity in periodontal health between African Americans and Mexican Americans? AB - OBJECTIVES: This study identified and contrasted prevalence and predictors of periodontitis among African Americans, Mexican Americans, and non-Hispanic Whites in the US adult population. DESIGN: Cross-sectional study. This study compared those with and without periodontitis in each racial/ethnic group. METHODS: This study was limited to records of US African-American, Mexican-American, and non Hispanic White adults at least 17 years of age who received a complete periodontal assessment as part of the dental examination in the third National Health and Nutrition Examination Survey (1988-1994). RESULTS: Despite the findings that Mexican Americans were less educated, poorer, and had less insurance coverage than African Americans, Mexican Americans had a similar prevalence of periodontitis as non-Hispanic Whites. African Americans had the highest prevalence among all groups. A similar scenario was observed in the multiple logistic analyses. CONCLUSIONS: Mexican Americans had periodontal health profiles closer to non-Hispanic Whites than did African Americans, despite the shared "minority" status. Public health research and practice should explicitly recognize that while "minorities" are considered a homogeneous group, they do not necessarily share the same health profiles. PMID- 11913614 TI - Treatment of heart failure in African Americans: clinical update. PMID- 11913615 TI - Community-based approaches with implications for hypertension control in blacks. PMID- 11913616 TI - Current guidelines in heart failure management. AB - 1. ACE inhibitors are indicated for Class I-IV HF due to LVSD. 2. ARBs are recommended for HF patients with contra-indications or intolerance to ACE inhibitors. 3. Spironolactone provides additional benefit for Class IV HF patients. 4. Beta-blockers (carvedilol or long-acting metoprolol) are indicated as additive treatment to an ACE inhibitor or ARB for Class II or III HF. 5. The beta-blocker carvedilol has been found to be safe and effective in patients with severe HF and appears to improve clinical outcomes in patients with LVSD post-MI. PMID- 11913617 TI - National MOTTEP: educating to prevent the need for transplantation. Minority Organ Tissue Transplant Education Program. AB - The purpose of this study was to determine whether participation in culturally appropriate, community-developed, health education programs addressing organ/tissue donation and preventing the need for transplantation, is an effective strategy for contributing to sustained behavior change among members of minority population groups. Within 15 target cities, 800 (40%) minority participants consented to a 2-3 month telephone follow-up survey. The 5-item questionnaire using items extracted from the pre- and post-survey instrument, addressed: 1) knowledge and beliefs; 2) willingness to sign donor cards and have family discussions; and 3) prevention/health behaviors. The preliminary findings based on telephone interviews with 253 participants indicate the following: [table: see text] The conclusions indicate that the community-based, culturally appropriate health education strategies used contributed to and sustained behavior changes. PMID- 11913618 TI - Homocysteine: an emerging risk factor. PMID- 11913619 TI - Environmental factors in the etiology of obesity in adolescents. AB - As the prevalence of obesity increases in children and adults from the United States and around the world, the interest in prevention is also increasing. For obesity prevention to have a chance to be successful, it should be based on evidences of risk factors rather than on empiric interventions. Selected risk factors for obesity present during adolescence are reviewed, such as television viewing, consumption of sugar-containing beverages, familial and socioeconomic environment, and recent changes in US society. The International Obesity Task Force's conceptual framework of environmental factors in the development of obesity is presented, as well as practical implications for obesity prevention and treatment in adolescence. PMID- 11913620 TI - Selective downregulation of endothelin-B receptors in diabetic African-American patients. AB - The purpose of this study was to investigate whether there are differences in the expression of the endothelin (ET) system in the peripheral vasculature of diabetic African-American (AA) and Caucasian (CA) patients. Tibial artery specimens were obtained from diabetic (MD = 8 and CAD = 5) and non-diabetic (AAND = 6 and CAND = 5) patients undergoing lower limb amputation. The gene expression of ET-1 precursor (PPET-1), ET(A)R and ET(B)R was determined by a reverse transcriptase polymerase chain reaction technique. PPET-1 and ET(A)R expression was up-regulated 4- and 3-fold, respectively, in both AA and CA diabetics (P<.05 vs non-diabetics). ET(B)R mRNA was significantly lower in AA diabetic patients. Function of ET-1 and ET receptors was assessed by vascular contractility assays. Vascular relaxation in response to sodium nitroprusside in arteries precontracted with ET-1 was significantly lower in AA (58% +/- 9) as compared to CA diabetics (74% +/- 5) (P<.05). In conclusion, this study demonstrated that the ET system is altered in favor of the contractile phenotype in AA diabetics and may contribute to the increased incidence of vascular complications in this population. PMID- 11913621 TI - Improving cardiovascular and renal outcome: maximizing drug therapy. Overview of stroke. PMID- 11913622 TI - Acute thrombolytic therapy in stroke: focus on the 3- to 6-hour time window. PMID- 11913623 TI - Racial differences in compliance with NCEP-II recommendations for secondary prevention at a Veterans Affairs medical center. AB - OBJECTIVE: The current NCEP-II guidelines recommend that secondary prevention patients should lower their LDL-cholesterol (LDL-C) below 100 mg/dL. We implemented a Lipid Management Program to aggressively achieve this goal. We report on the impact of this intervention on compliance rates for African Americans (AA) vs Whites (W) treated with an HMG-Co-A reductase inhibitor for secondary prevention at the veterans affairs hospital. METHODS: We reviewed all patients with coronary artery disease (CAD) and/or diabetes mellitus (DM) at our institution on monotherapy with an HMG-Co-A reductase inhibitor in 1999. We examined the initial and post intervention lipid profiles for both races. RESULTS: The groups differed in that compared to the Whites, the AA were younger (65.8 vs 71.4, P = .0001); had a higher prevalence of type 2 DM (70.1% vs 40.8%, P = .001), had more obesity (57.5% vs 41.0%, P = .001), and were more likely to smoke (42.5% vs 9.6%, P = .001). AA had more clinic visits (5.04/pt vs 3.95/pt, P = .0001) and fasting lipid profiles (4.46/pt vs 3.0/pt, P = .0001). There was no difference in the prevalence of hypertension or HMG-CoA reductase inhibitor dose. AA were less likely to achieve the goal for LDL-C recommended by NCEP-II (40.94% vs 56.9%, P = .001). CONCLUSION: Despite equivalent doses of statin, AA were less likely to meet NCEP-II recommendations. This occurred even though AA had more clinic visits and lipid profiles. Our intervention did not narrow this racial gap in compliance rates. Possible explanations include: 1) variations in patient compliance; 2) impact of differences in lifestyle (DM, obesity, and smoking); and 3) the need for more intensive drug therapy in patients starting with a higher baseline LDL-C. PMID- 11913625 TI - Diet- and blood pressure-related knowledge, attitudes, and hypertension prevalence among African Americans: the KDBP Study. Knowledge of Diet and Blood Pressure. AB - Despite attempts to raise awareness of the effect lifestyle factors such as diet have on health, little is known about the results of such efforts among populations at highest risk for hypertension (HTN). The Knowledge of Diet and Blood Pressure (KDBP) Study investigated the relationship between diet- and blood pressure-related knowledge and HTN prevalence, and attitudes among a church-based population of African Americans. One hundred ninety-six adults were randomly selected from 6 churches in an urban area of North Carolina. After study criteria and missing data exclusions, the study sample comprised 179 individuals. A knowledge index assessed awareness of: the definition of HTN; its risk and prognostic factors; risk-preventing and risk-promoting nutrition; and related nutritional recommendations (score range: 0-100, 100 = most knowledgeable). Health attitudes were assessed primarily through Likert-type questions. HTN was defined as SBP > or = 140 mm Hg, DBP > or = 90 mm Hg, or anti hypertensive medication use. The mean SBP and DBP were 135.4 +/- 21.6 mm Hg and 78.8 +/- 15.7 mm Hg, respectively, and 55.9% of participants had HTN. The mean knowledge score was 76.1 (+/- 10.6). There was no statistically significant difference in mean knowledge score by HTN status (known HTN: 76.9 [+/- 11.31); unknown HTN (ie, the participant was unaware of the presence of HTN): 76.1 (+/- 9.3); no HTN: 75.3 (+/ 10.4), P = .665). Attitudes were not significantly related to knowledge and HTN prevalence, despite apparent trends. However, logistic regression analyses revealed that age, occupation, and church site were significant correlates of this relationship. Further exploration of attitudes and socioeconomic factors in assessing health awareness and HTN prevalence in this population is recommended. PMID- 11913624 TI - Association between angiotensin II type I receptor polymorphism and resting hemodynamics in black and white youth. AB - OBJECTIVES: An angiotensin II type I receptor polymorphism (AGTR1/A1166C) was previously found to be associated with hypertension. The purpose of this study was to investigate the relationship between this polymorphism and resting measures of hemodynamics in normotensive youth. DESIGN: Subjects were 41 Whites (mean +/- SD: 18 +/- 3 y old, 26 males) and 73 Blacks (19 +/- 2 y old, 55 males) with a positive family history of hypertension. METHODS: Hemodynamic measures included resting systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR). RESULTS: Allele frequencies were significantly different (chi2 = 14.10, P < or = .001) between Whites and Blacks (.23 vs .06 for the C allele, respectively). For all subsequent analyses, subjects were categorized into two genotype groups, carriers and non-carriers of the C allele, because only two Whites and no Blacks were homozygous for the C allele. Genotype frequencies were significantly different (chi2 = 12.66, P < or = .0011) between Whites and Blacks (.41 vs .12 for the carriers, respectively). Among Whites, univariate analyses of covariance, using body mass index and age as covariates, indicated that carriers of the C allele compared to non-carriers, had a higher DBP (61.6 +/ 6.7 vs 57.8 +/- 6.2 mm Hg, P < or = .05) and HR (68.0 +/- 10.5 vs 61.1 +/- 8.1, P < or = .05). Genotype was not associated with resting hemodynamic measures in Blacks (all P values > .05). CONCLUSIONS: These results are consistent with findings that have typically involved White adults, and demonstrate that the renin system does not seem to play as great a role in BP control in Blacks as it does in Whites. PMID- 11913626 TI - Changes in overweight in youth over a period of 7 years: impact of ethnicity, gender and socioeconomic status. AB - OBJECTIVE: To examine changes in overweight in youth over a period of 7 years, within the context of ethnicity, gender, and socioeconomic status (SES). METHODS: Participants in a longitudinal study of cardiovascular (CV) risk factor development were evaluated on 2 occasions an average of 7.2 +/- 0.5 years apart. There were 253 subjects (121 Blacks, 132 Whites, 130 females) with a mean age of 8.8 +/- 2.0 years at the initial visit. SES was determined by Hollingshead Social Status Index and was categorized as low, middle (+/-1 standard deviation from mean), and high. Overweight was defined as >85th percentile in Body Mass Index (BMI) for age and gender (National Health and Nutrition Examination Survey [NHANES] norms). RESULTS: Standardized BMI increased significantly over the course of the study (P<.001). The increase was not significantly different by ethnicity or by gender (both P values>.05) but did significantly differ by SES (P<.001). Overall, the prevalence of overweight increased from 31% to 40% (P<.001); however, for those with low SES, the prevalence increased from 37% to 67% (P<.001). CONCLUSION: Lower SES youth, regardless of ethnicity or gender, exhibited extremely large increases in general adiposity over time and are at particular risk for development of a number of obesity-related problems, such as hypertension. Primary prevention of obesity is needed in youth, particularly among those from low SES backgrounds. PMID- 11913627 TI - The impact of racism on health. PMID- 11913628 TI - The impact of socioeconomic status on health disparities in the United States. PMID- 11913629 TI - Diabetes and obesity. Epidemiology and the role of prevention: what do we know? PMID- 11913631 TI - Foreword: Building a healthy South--the challenge is now. PMID- 11913630 TI - Current trends in obesity: clinical impact and interventions that work. PMID- 11913632 TI - Diabetes: update on management and therapy. PMID- 11913633 TI - Mental health: the gap between knowledge and practice in the community--how can we help? PMID- 11913634 TI - Evaluating the patient with early dementia. PMID- 11913635 TI - Closing the gap: strategies for increasing immunization levels among at-risk populations. PMID- 11913636 TI - Hepatitis update: the role of primary care practitioners in prevention, treatment, and vaccination. PMID- 11913637 TI - Influenza and pneumococcal disease: does vaccination make a difference? PMID- 11913638 TI - Cardiovascular health. Cardiovascular epidemiology and the role of prevention: what do we know and what must we do? PMID- 11913639 TI - Introduction: Achieving optimal outcomes in primary care. PMID- 11913640 TI - What is new in cardiovascular health? PMID- 11913641 TI - Hypertension: treatment--now & tomorrow. PMID- 11913642 TI - Congestive heart failure: improving outcomes for at-risk populations. PMID- 11913643 TI - HIV and AIDS: an update on impact in the United States and in the Department of Health and Human Services Region IV. PMID- 11913644 TI - HIV and AIDS. An overview of prevention and treatment and one approach to fighting the epidemic that works: "SAFE". PMID- 11913645 TI - Sexually transmitted disease. risk, prevention, and treatment: an update. PMID- 11913646 TI - Tuberculosis: current epidemiology and recommendations for treatment of TB infection. PMID- 11913647 TI - Cancer. Populations at risk: who is dying and why? PMID- 11913648 TI - Cancer prevention and screening: do they make a difference? PMID- 11913649 TI - Prostate cancer. PMID- 11913650 TI - Lung cancer in African Americans. PMID- 11913651 TI - Colon cancer. PMID- 11913652 TI - Breast cancer. PMID- 11913653 TI - First plenary session: Healthy people 2010--eliminating health disparities culture, community, and primary care. PMID- 11913654 TI - Depression in the primary care setting. PMID- 11913655 TI - Adaptive constraints and the phylogenetic comparative method: a computer simulation test. AB - Recently, the utility of modern phylogenetic comparative methods (PCMs) has been questioned because of the seemingly restrictive assumptions required by these methods. Although most comparative analyses involve traits thought to be undergoing natural or sexual selection, most PCMs require an assumption that the traits be evolving by less directed random processes, such as Brownian motion (BM). In this study, we use computer simulation to generate data under more realistic evolutionary scenarios and consider the statistical abilities of a variety of PCMs to estimate correlation coefficients from these data. We found that correlations estimated without taking phylogeny into account were often quite poor and never substantially better than those produced by the other tested methods. In contrast, most PCMs performed quite well even when their assumptions were violated. Felsenstein's independent contrasts (FIC) method gave the best performance in many cases, even when weak constraints had been acting throughout phenotypic evolution. When strong constraints acted in opposition to variance generating (i.e., BM) forces, however, FIC correlation coefficients were biased in the direction of those BM forces. In most cases, all other PCMs tested (phylogenetic generalized least squares, phylogenetic mixed model, spatial autoregression, and phylogenetic eigenvector regression) yielded good statistical performance, regardless of the details of the evolutionary model used to generate the data. Actual parameter estimates given by different PCMs for each dataset, however, were occasionally very different from one another, suggesting that the choice among them should depend on the types of traits and evolutionary processes being considered. PMID- 11913656 TI - Patterns of divergence in the effects of mating on female reproductive performance in flour beetles. AB - Sexual selection can lead to rapid divergence in reproductive characters. Recent studies have indicated that postmating events, such as sperm precedence, may play a key role in speciation. Here, we stress that other components of postmating sexual selection may be involved in the evolution of reproductive isolation. One of these is the reproductive investment made by females after mating (i.e., differential allocation). We performed an experiment designed to assess genetic divergence in the effects of mating on female reproductive performance in flour beetles, Tribolium castaneum. Females were mated to males of three different wild type genotypes at two different frequencies, in all possible reciprocal combinations. Male genotype affected all aspects of female reproduction, through its effects on female longevity, total offspring production, reproductive rate, mating rate, and fertility. Moreover, male and female genotype interacted in their effects on offspring production and reproductive rate. We use the pattern of these interactions to discuss the evolutionary process of divergence and suggest that the pattern is most consistent with that expected if divergence was driven by sexually antagonistic coevolution. In particular, the fact that females exhibited a relatively weak response to males with which they were coevolved suggests that females have evolved resistance to male gonadotropic signals/stimuli. PMID- 11913657 TI - A bird's-eye view of the C-value enigma: genome size, cell size, and metabolic rate in the class aves. AB - For half a century, variation in genome size (C-value) has been an unresolved puzzle in evolutionary biology. While the initial "C-value paradox" was solved with the discovery of noncoding DNA, a much more complex "C-value enigma" remains. The present study focuses on one aspect of this puzzle, namely the small genome sizes of birds. Significant negative correlations are reported between resting metabolic rate and both C-value and erythrocyte size. Cell size is positively correlated with both nucleus size and C-value in birds, as in other vertebrates. These findings shed light on the constraints acting on genome size in birds and illustrate the importance of interactions among various levels of the biological hierarchy, ranging from the subchromosomal to the ecological. Following from a discussion of the mechanistic bases of the correlations reported and the processes by which birds achieved and/or maintain small genomes, a pluralistic approach to the C-value enigma is recommended. PMID- 11913658 TI - Heterosis and outbreeding depression in descendants of natural immigrants to an inbred population of song sparrows (Melospiza melodia). AB - We studied heterosis and outbreeding depression among immigrants and their descendants in a population of song sparrows on Mandarte Island, Canada. Using data spanning 19 generations, we compared survival, seasonal reproductive success, and lifetime reproductive success of immigrants, natives (birds with resident-hatched parents and grandparents), and their offspring (F1s, birds with an immigrant and a native parent, and F2s, birds with an immigrant grandparent and resident-hatched grandparent in each of their maternal and paternal lines). Lifetime reproductive success of immigrants was no worse than that of natives, but other measures of performance differed in several ways. Immigrant females laid later and showed a tendency to lay fewer clutches, but had relatively high success raising offspring per egg produced. The few immigrant males survived well but were less likely to breed than native males of the same age that were alive in the same year. Female F1s laid earlier than expected based on the average for immigrant and native females, and adult male F1s were more likely to breed than expected based on the average for immigrant and native males. The performance differences between immigrant and native females and between F1s and the average of immigrants and natives are consistent with the hypothesis that immigrants were disadvantaged by a lack of site experience and that immigrant offspring benefited from heterosis. However, we could not exclude the possibility that immigrants had a different strategy for optimizing reproductive success or that they experienced ecological compensation for life-history parameters. For example, the offspring of immigrants may have survived well because immigrants laid later and produced fewer clutches, thereby raising offspring during a period of milder climatic conditions. Although sample sizes were small, we found large performance differences between F1s and F2s, which suggested that either heterosis was associated with epistasis in F1s, that F2s experienced outbreeding depression, or that both phenomena occurred. These findings indicate that the performance of dispersers may be affected more by fine-scale genetic differentiation than previously assumed in this and comparable systems. PMID- 11913659 TI - The ecology and genetics of fitness in Chlamydomonas. VIII. The dynamics of adaptation to novel environments after a single episode of sex. AB - According to classical evolutionary theory, sexual recombination can generate the variation necessary to adapt to changing environments and thereby confer an evolutionary advantage of sexual over asexual reproduction. Using the green alga, Chlamydomonas reinhardtii, we investigated the effect of a single sexual episode on adaptation of heterotrophic growth on different carbon sources. In an initial mixture of isolates, sex was induced and the resulting offspring constituted the sexual populations, along with any unmated vegetative cells; the unmated mixture of isolates represented the asexual populations. Mean and variance in division rates (i.e., fitness) were measured four times during approximately 50 generations of vegetative growth in the dark on all possible combinations of four carbon sources. Consistent with effects of recombination of epistatic genes in linkage disequilibrium, sexual populations initially had a higher variance in fitness, but their mean fitness was lower than that of asexual populations, possibly due to recombinational load. Subsequently, fitness of sexual populations exceeded that of asexual ones, but finally they regained parity in both mean and variance of fitness. Although recombination was not more effective on more complex substrates, these results generally support the idea that sex can accelerate adaptation to novel environments. PMID- 11913660 TI - Time to the most recent common ancestor and divergence times of populations of common chaffinches (Fringilla coelebs) in Europe and North Africa: insights into Pleistocene refugia and current levels of migration. AB - We analyzed sequences from a 275-bp hypervariable region in the 5' end of the mitochondrial DNA control region in 190 common chaffinches (Fringilla coelebs) from 19 populations in Europe and North Africa, including new samples from Greece and Morocco. Coalescent techniques were applied to estimate the time to the most recent common ancestor (TMRCA) and divergence times of these populations. The first objective of this study was to infer the locations of refugia where chaffinches survived the last glacial episode, and this was achieved by estimating the TMRCA of populations in regions surrounding the Mediterranean that were unglaciated in the late Pleistocene. Although extant populations in Iberia, Corsica, Greece, and North Africa harbor haplotypes that are basal in a phylogenetic tree, this information alone cannot be used to infer that these localities served as refugia, because it is impossible to infer the ages of populations and their divergence times without also considering the population genetic processes of mutation, migration, and drift. Provided we assume the TMRCAs of populations are a reasonable estimate of a population's age, coalescent based methods place resident populations in Iberia, Corsica, Greece, and North Africa during the time of the last glacial maximum, suggesting these regions served as refugia for the common chaffinch. The second objective was to determine when populations began diverging from each other and to use this as a baseline to estimate current levels of gene flow. Divergence time estimates suggest that European populations began diverging about 60,000 years before present. The relatively recent divergence of populations in North Africa, Italy, and Iberia may explain why classic migration estimates based on equilibrium assumptions are high for these populations. We compare these estimates with nonequilibrium-based estimates and show that the nonequilibrium estimates are consistently lower than the equilibrium estimates. PMID- 11913661 TI - Testing for genetic evidence of population expansion and contraction: an empirical analysis of microsatellite DNA variation using a hierarchical Bayesian model. AB - The role of past climatic change in shaping the distributions of tropical rain forest vertebrates is central to long-standing hypotheses about the legacy of the Quaternary ice ages. One approach to testing such hypotheses is to use genetic data to infer the demographic history of codistributed species. Population genetic theory that relates the structure of allelic genealogies to historical changes in effective population size can be used to detect a past history of demographic expansion or contraction. The fruit bats Cynopterus sphinx and C. brachyotis (Chiroptera: Pteropodidae) exhibit markedly different distribution patterns across the Indomalayan region and therefore represent an exemplary species pair to use for such tests. The purpose of this study was to test alternative hypotheses about historical patterns of demographic expansion and contraction in C. sphinx and C. brachyotis using a coalescent-based analysis of microsatellite variation. Specifically, we used a hierarchical Bayesian model based on Markov chain Monte Carlo simulations to estimate the posterior distribution of genealogical and demographic parameters. The results revealed strong evidence for population contraction in both species. Evidence for a population contraction in C. brachyotis was expected on the basis of biogeographic considerations. However, similar evidence for population contraction in C. sphinx does not support the hypothesis that this species underwent a pronounced range expansion during the late Quaternary. Genetic evidence for population decline may reflect the consequences of habitat destruction on a more recent time scale. PMID- 11913662 TI - A comparative study of leukocyte counts and disease risk in primates. AB - Little is known about how the risk of disease varies across species and its consequences for host defenses, including the immune system. I obtained mean values of basal white blood cells (WBC) from 100 species of primates to quantify disease risk, based on the assumption that higher baseline WBC counts will be found in species that experience greater risk of acquiring infectious disease. These data were used to investigate four hypotheses: disease risk is expected to increase with (1) group size and population density; (2) greater contact with soil-borne pathogens during terrestrial locomotion; (3) a slow life history; and (4) increased mating promiscuity. After controlling for phylogeny, WBC counts increased with female mating promiscuity, as reflected in discrete categories of partner number, relative testes mass, and estrous duration. By comparison, the social, ecological, and life-history hypotheses were unsupported in comparative tests. In terms of confounding variables, some WBC types were associated with body mass or activity period, but these variables could not account for the association with mating promiscuity. Several factors may explain why hypotheses involving social, ecological, and life-history factors went unsupported in these tests, including the role of behavioral counterstrategies to disease, restrictions on female choice of mating partners, and the effect of transmission mode on parasite strategies and host defenses. PMID- 11913663 TI - Molecular correlates of reproductive isolation. AB - Evolution of reproductive isolation as a byproduct of genetic divergence in isolated populations is the dominant (albeit not exclusive) mode of speciation in sexual animals. But little is known about the factors linking speciation to general divergence. Several authors have argued that allopatric speciation should proceed more rapidly if isolated populations also experience divergent selection. Reproductive isolation between allopatric populations is not subject to direct selection; it can accumulate only by random drift or as a fortuitous byproduct of selection on other traits. Here I present a novel analysis of published data, demonstrating that pre- and postmating isolation of Drosophila species are more tightly correlated with allozyme divergence than with silent DNA divergence. Inasmuch as proteins are more subject to the action of natural selection than are silent DNA polymorphisms, this result provides broad support for a model of selection-mediated allopatric speciation. PMID- 11913664 TI - Obligate brood parasites as selective agents for evolution of egg appearance in passerine birds. AB - Many passerine host species have counteracted the parasite egg mimicry in their coevolutionary arms race with the common cuckoo (Cuculus canorus) by evolving increased interclutch and reduced intraclutch variation in egg appearance. Such variations make it easier for hosts to recognize a foreign egg, reduce the possibility of making recognition errors, and reduce the ability of the cuckoo to mimic the eggs of a particular host. Here, we investigate if such clutch characteristics have evolved among North American passerines. We predict that due to the absence of brood parasites with egg mimicry on this continent, these passerines should (1) not show any relationship between rejection rates and intra or interclutch variation, and (2) intraclutch variation should be lower and interclutch variation higher in European hosts exposed to cuckoo parasitism as compared to North American hosts parasitized by cowbirds. Here we present data that show support for most of these and other predictions, as well as when controlling statistically for effects of common descent. However, the effect of continent on intraclutch variation was less than predicted and we discuss a possible reason for this. All things considered, the results demonstrate that parasitism by a specialist brood parasite with egg mimicry is a powerful selective force regarding the evolution of egg characteristics in passerine birds. PMID- 11913665 TI - Inbreeding alters resistance to insect herbivory and host plant quality in Mimulus guttatus (Scrophulariaceae). AB - Previous studies have demonstrated genetic variation for resistance to insect herbivores and host plant quality. The effect of plant mating system, an important determinant of the distribution of genetic variation, on host plant characteristics has received almost no attention. This study used a controlled greenhouse experiment to examine the effect of self- and cross-pollination in Mimulus guttatus (Scrophulariaceae) on resistance to and host plant quality for the xylem-feeding spittlebug Philaenus spumarius (Homoptera: Cercopidae). Spittlebugs were found to have a negative effect on two important fitness components in M. guttatus, flower production and above ground biomass. One of two M. guttatus populations examined showed a significant interaction between the pollination and herbivore treatments. In this case, the detrimental effects of herbivores on biomass and flower production were much more pronounced in inbred (self) plants. The presence of spittlebug nymphs increased inbreeding depression by as much as three times. Pollination treatments also had significant effects on important components of herbivore fitness, but these effects were in opposite directions in our two host plant populations. Spittlebug nymphs maturing on self plants emerged as significantly larger adults in one of our host plant populations, indicating that inbreeding increased host plant quality. In our second host plant population, spittlebug nymphs took significantly longer to develop to adulthood on self plants, indicating that inbreeding decreased host plant quality. Taken together these results suggest that the degree of inbreeding in host plant populations can have important and perhaps complex effects on the dynamics of plant-herbivore interactions and on mating-system evolution in the host. PMID- 11913666 TI - Phylogeny of Gaertnera Lam. (Rubiaceae) based on multiple DNA markers: evidence of a rapid radiation in a widespread, morphologically diverse genus. AB - Phylogenetic relationships among 28 of the 68 species of the paleotropical genus Gaertnera (Rubiaceae) and two related genera were inferred from nucleotide sequence variation in four nuclear DNA (nDNA) markers: the internal transcribed spacers of nuclear rDNA (ITS), the large and small copies of phosphoenolpyruvate carboxylase (PepC-large and PepC-small), and triose phosphate isomerase (Tpi). Phylogenetic analysis of the combined nDNA dataset suggested that Gaertnera is monophyletic, but genetic variation among species was insufficient to reconstruct well-supported relationships within the genus. This was counter to expectations based on the very distinct morphologies and widespread distribution of the genus (West Africa to Sulawesi). Molecular clock analyses suggested variable dates of origin for Gaertnera depending upon the calibration method used. The most plausible calibration implies that Gaertnera migrated to Africa during the early Tertiary, possibly via a boreotropical land bridge and suggests that Gaertnera started to radiate 5.21 +/- 0.14 million years ago. This implies that range expansion in the group has occurred via a number of long-distance dispersal events rather than vicariance. The molecular clock estimate in turn estimated an unusually rapid lineage diversification rate within the radiation of 0.717-0.832 species/million years, comparable to those estimated for radiations on oceanic islands. Although low interspecific competition levels may have contributed to the diversification of Gaertnera on Mauritius, the mechanisms driving the rapid radiation of the group in other parts of its range remain elusive. PMID- 11913667 TI - Episodic chromosomal evolution in Planipapillus (Onychophora: Peripatopsidae): a phylogenetic approach to evolutionary dynamics and speciation. AB - Planipapillus, a clade of onychophorans from southeastern Australia, exhibits substantial chromosomal variation. In the context of a robust phylogeny based on nuclear and mitochondrial sequence data, we evaluate models of chromosomal evolution and speciation that differ in the roles assigned to selection, mutation, and drift. Permutation tests suggest that all chromosome rearrangements in the clade have been centric fusions and, on the basis of parsimony and maximum likelihood methods with independent estimates of branch lengths, we conclude that at least 31 centric fusions have been fixed in Planipapillus. A likelihood-ratio test approach, which is independent of our point estimates of ancestral states, rejects an evolutionary model in which the mutation rate is constant and centric fusions are effectively neutral. In contrast to the nucleotide sequence data, which are consistent with neutrality and rate constancy, centric fusions in Planipapillus are underdominant, spontaneous fusion rates vary among lineages, or both. We predict an inverse relationship between rates of chromosomal evolution and historical population size. Chromosomal evolution may play a role in speciation in Planipapillus, both by interactions between centric fusions with monobrachial homology and by the accumulation of multiple weakly underdominant fusions. PMID- 11913668 TI - The evolution of trade-offs: testing predictions on response to selection and environmental variation. AB - The concept of phenotypic trade-offs is a central element in evolutionary theory. In general, phenotypic models assume a fixed trade-off function, whereas quantitative genetic theory predicts that the trade-off function will change as a result of selection. For a linear trade-off function selection will readily change the intercept but will have to be relatively stronger to change the slope. We test these predictions by examining the trade-off between fecundity and flight capability, as measured by dorso-longitudinal muscle mass, in four different populations of the sand cricket, Gryllus firmus. Three populations were recently derived from the wild, and the fourth had been in the laboratory for 19 years. We hypothesized that the laboratory population had most likely undergone more and different selection from the three wild populations and therefore should differ from these in respect to both slope and intercept. Because of geographic variation in selection, we predicted a general difference in intercept among the four populations. We further tested the hypothesis that this intercept will be correlated with proportion macropterous and that this relationship will itself vary with environmental conditions experienced during both the nymphal and adult period. Observed variation in the phenotypic trade-off was consistent with the predictions of the quantitative genetic model. These results point to the importance of modeling trade-offs as dynamic rather than static relationships. We discuss how phenotypic models can incorporate such variation. The phenotypic trade-off between fecundity and dorso-longitudinal muscle mass is determined in part by variation in body size, illustrating the necessity of considering trade offs to be multi factorial rather than simply bivariate relationships. PMID- 11913669 TI - Quantitative genetics of growth and development time in the burying beetle Nicrophorus pustulatus in the presence and absence of post-hatching parental care. AB - Despite a growing interest in the evolutionary aspects of maternal effects, few studies have examined the genetic consequences of maternal effects associated with parental care. To begin to provide data on nonlaboratory or nondomestic animals, we compared the effect of presence and absence of parental care on phenotype expression of larval mass and development time at different life history stages in the burying beetle Nicrophorus pustulatus. This beetle has facultative care; parents can feed their larvae through regurgitation of digested carrion or offspring can feed by themselves from previously prepared carrion. To investigate larval responses to these two levels of care, including estimates of additive genetic effects, maternal effects, and genotype-by-environment interactions, we used a half-sibling split-family breeding experiment-raising half of the offspring of a family in the presence of their mother and the other half without their mother present. Larvae reared with their mother present were on average heavier and developed faster, although some of the differences in development decreased or were eliminated by the adult stage. These results suggest that presence or absence of post-hatching maternal care plays an important role in phenotype expression early in life, whereas later the phenotype of the offspring is determined mainly by the genotype and/or unshared environmental effects. Our study also permitted us to examine the differences in genetic effects between the two care environments. Heritabilities, maternal/common environment effect, and most genetic correlations did not differ between the care treatments. Genetic analyses revealed substantial additive genetic effects for development time but small effects for measures of body mass. Maternal plus common environment effects were high for measures of mass but low for development time, suggesting that indirect genetic effects of maternal and/or common environment are less important for the evolution of development time than for mass. Estimates of genetic correlations revealed a trade-off between the duration of the two development stages after the offspring left the carrion. There was also a negative genetic correlation between the time spent on carrion and the mass at 72 h, when mothers usually stop feeding. The analysis of genotype by-environment interactions indicates substantial variation among maternal families in response to care. Presence or absence of parental care may therefore contribute to the additive genetic variance through its interaction with the maternal component of the additive genetic variance. The presence of this interaction further suggests that parents may vary in care strategies, with some parents dispersing after preparation of the carrion and some parents staying with the larvae. This interaction may help maintain genetic variation in growth, development time, and parental care behavior. Additional work is needed, however, to quantify indirect genetic effects and genetic variation in parental care behavior itself. PMID- 11913670 TI - Movement disorders associated with atypical antipsychotic drugs. AB - Data from clinical trials reviewed in this article fulfill predictions based on preclinical findings that atypical antipsychotic drugs are associated with a reduced potential for inducing extrapyramidal symptoms (EPS) and other movement disorders. Atypical drugs have been shown to reduce all subtypes of acute EPS, the frequency of EPS-related patient dropouts, and the need for concomitant antiparkinsonian drug use. Clozapine remains superior to other atypicals in treating psychosis without worsening motor symptoms in patients with Parkinson's disease. Atypicals may be selectively advantageous in treating schizophrenic patients with a predisposition to catatonia. Although the risk of developing lethal neuroleptic malignant syndrome may be diminished with atypical drugs, clinicians must remain alert to the signs of this disorder. Atypicals have reduced liability for inducing tardive dyskinesia (TD) and show antidyskinetic properties in patients with preexisting TD. Passive resolution of TD may be facilitated in some patients by the use of these agents. Thus, the risk of movement disorders has become only one of several considerations in choosing among antipsychotic drugs. PMID- 11913671 TI - Cardiovascular effects of antipsychotics used in bipolar illness. AB - Many antipsychotics are used to treat disorders other than schizophrenia, such as bipolar disorder. However, some of these agents are associated with cardiovascular side effects that can have serious or fatal implications for the patient. One dangerous side effect is QT prolongation, which can lead to torsades de pointes and ventricular arrhythmia. The group of drugs labeled as antipsychotics is diverse, making it challenging for the clinician to keep track of which agents cause which effects and important for the clinician to learn to identify signs of cardiovascular side effects and manage those events when they occur. PMID- 11913672 TI - Medical management of obesity associated with mental disorders. AB - Obesity and mental disorders are major public health problems that co-occur to a significant degree. They also significantly overlap in phenomenology and response to medications. However, many psychotropic agents have adverse effects on appetite, binge eating, and weight. In this review, we aim to improve understanding of the relationship between obesity and mental illness and provide practical clinical guidelines for the management of obesity associated with mental disorders. PMID- 11913673 TI - Clinical pharmacodynamics and pharmacokinetics of antimanic and mood-stabilizing medications. AB - BACKGROUND: A number of medications are now available or are in development as antimanic and/or mood-stabilizing agents. We reviewed the clinical pharmacodynamics and pharmacokinetics of these agents to provide a summary of these properties relevant to clinical practice. METHOD: We conducted a MEDLINE search augmented by a manual search of bibliographies and a review of textbooks to identify articles regarding the clinical pharmacology of lithium, valproate, carbamazepine, oxcarbazepine, olanzapine, clozapine, risperidone, ziprasidone, quetiapine, lamotrigine, and topiramate. RESULTS: Not surprisingly, there are a number of clinically relevant pharmacodynamic and pharmacokinetic differences among these medications, and these differences are discussed. CONCLUSION: Knowledge of the clinical pharmacology of established and putative antimanic and mood-stabilizing medications is important in administering these agents safely and effectively. PMID- 11913674 TI - Management of weight gain in patients with schizophrenia. AB - Of the roughly 55% of the United States population that is considered overweight, half meet the criteria for obesity. Obesity is associated with serious health risks, but many clinicians graduate from medical school without a clear understanding of the effects of the foods that they and their patients consume. Obesity is more prevalent in people with mental illnesses, which poses an even greater challenge to clinicians. Antipsychotic treatment can cause weight gain, and mentally ill patients generally lack an understanding of nutrition as well as the ability to afford healthier foods. Therefore, clinicians must educate themselves about appropriate measures for preventing weight gain before or immediately after initiating antipsychotic therapy. Strategies for weight gain management that have proven effective in clinical trials include regular check ups, lifestyle and medication counseling, medication assessments, behavioral control programs, and pharmacologic intervention. These approaches are necessary for clinicians to consider if efforts at reintegration of mentally ill patients are to succeed. PMID- 11913675 TI - Metabolic side effects of antipsychotics: focus on hyperglycemia and diabetes. AB - Approximately 16 million people in the United States have diabetes, and the World Health Organization has estimated that the worldwide prevalence of diabetes will more than double from 1995 to 2025. Type 2 diabetes, the most common form of diabetes, may be 2 to 4 times more prevalent in patients with severe mental disorders. Within the psychiatric community, there is a great deal of concern about diabetes as a potential side effect of antipsychotic agents. An important population to remember in the context of treatment-emergent hyperglycemia is the 20 million people with impaired glucose tolerance (IGT) (fasting plasma glucose > 110 mg/dL and < 126 mg/dL or 2-hour postload glucose > 140 mg/dL and < 200 mg/dL according to the American Diabetes Association). This prediabetic condition has a 5% to 10% annual risk of converting to diabetes. One hypothesis for antipsychotic treatment-emergent diabetes during double-blind, randomized, controlled trials is that people who develop diabetes soon after the initiation of drug therapy for schizophrenia may have had undiagnosed IGT or diabetes before they started treatment. The emergence of diabetes in clinical practice may be due to an observation effect, but because the incidence of diabetes is greater in people with severe mental illnesses, it is crucial for psychiatrists to be aware of national guidelines for the diagnosis and treatment of type 2 diabetes. PMID- 11913676 TI - The reproductive safety profile of mood stabilizers, atypical antipsychotics, and broad-spectrum psychotropics. AB - There has been growing concern about the potential iatrogenic effects of several newer psychotropic drugs on reproductive health safety in women. Areas of particular concern in this regard include (1) controversies about a potential association between the use of valproate and development of polycystic ovary syndrome (PCOS), (2) the safety of use of newer psychotropic medications during pregnancy, and (3) safety issues with these medications in women while breastfeeding. This review summarizes current information about each of these areas. In particular, existing data suggest that (1) PCOS very likely represents a complex neuroendocrine disorder with multiple determinants; (2) menstrual irregularities may be a frequently seen phenomenon in women with bipolar illness, at least partially independent of psychotropic drug therapy; (3) potential central nervous system teratogenicity remains substantial during first-trimester exposure to valproate or carbamazepine; (4) with newer agents used for bipolar disorder and schizophrenia, safety data during pregnancy, while not definitive, are most abundant with olanzapine and with lamotrigine; relatively less is known about systematic pregnancy outcomes with other atypical antipsychotics or newer anticonvulsants; and (5) risks for neonatal safety during lactation continue to appear substantial with lithium, are of potential concern with lamotrigine and clozapine, are quite likely minimal with valproate or carbamazepine, and are indeterminate with most other new anticonvulsants or atypical antipsychotics. Recommendations are presented for clinical management in each of these instances. PMID- 11913677 TI - Prolactin elevation with antipsychotic medications: mechanisms of action and clinical consequences. AB - Antipsychotic agents differ in efficacy and side effects such as movement disorders and prolactin elevation because of varying mechanisms of action. A revised nomenclature for antipsychotic agents, which categorizes the drugs according to efficacy, risk of movement disorders, and risk of prolactin elevation, is described. Prolactin elevation, a potential side effect of some antipsychotic medications, is underdiagnosed but can have serious short-term and long-term consequences. Short-term problems include menstrual irregularities, sexual dysfunction, and depression. Long-term problems related to prolactin elevation include decreased bone density and osteoporosis, relapse of psychosis because of poor compliance due to sexual dysfunction or depression, and perhaps cancer, although more research in this area is needed. Despite the serious nature of these effects, prolactin elevation is seldom detected because clinicians often fail to inquire about sexual function or other symptoms that signal that a patient's prolactin may be elevated. These are problems that patients may not bring up with clinicians unless they are asked. Therefore, when patients are taking antipsychotic medications, clinicians should regularly inquire about sexual dysfunction, depression, menstrual disturbances, galactorrhea, and gynecomastia. PMID- 11913678 TI - Membranous tracheal rupture in children following minor blunt cervical trauma. AB - Injuries to the tracheobronchial tree are well-recognized sequelae of massive blunt or penetrating injuries of the neck or chest. They may also occur as a rare complication of endotracheal intubation. We present 2 cases of a less well recognized clinical entity, rupture of the membranous trachea following minimal blunt trauma to the neck in children. The case histories and management of this disorder are discussed. Recognition and treatment of this problem requires a high index of suspicion for the lesion and timely investigations. Open repair of the trachea to secure a stable airway is recommended for this injury, unless the wound is small and the wound edges are well approximated. PMID- 11913679 TI - Metachronous second primary cancers after successful partial laryngectomy for invasive squamous cell carcinoma of the true vocal cord. AB - The current retrospective studies documented the incidence, sites of occurrence, risk factors, and outcome of metachronous second primary cancers (MSPCs) among an inception cohort of 410 patients with invasive squamous cell carcinoma of the true vocal cord successfully treated with partial laryngectomy at a single institution. The Kaplan-Meier actuarial life-table method was used to document the relationship between the incidence of MSPCs and survival data. Univariate analysis was performed for potential statistical relationships among the incidence of MSPCs, the site of occurrence, and different variables. The overall incidence of MSPCs was 23.9% (98/410). The 10-year actuarial survival estimate for MSPCs was 20.4%. The incidence curve of MSPCs was linear, resulting in a 2%/y rate of development for MSPCs. In univariate analysis, the only variable that demonstrated a statistical correlation with the incidence of MSPCs was smoking, with MSPCs being statistically more likely to occur in smokers than in nonsmokers (p = .04). The main sites of origin for MSPCs were the lung (25.5% of cases), other non-upper aerodigestive tract sites (32.7%), and the upper aerodigestive tract (41.8%). The 10-year actuarial estimates for MSPCs were 9.1% in the upper aerodigestive tract, 7.1% in sites other than the lung or upper aerodigestive tract, and 6.6% in the lung. The incidence curve for MSPCs was linear, whatever the site of origin, resulting in 1 %/y, 0.7%/y, and 0.6%/y rates of development for MSPCs in the upper aerodigestive tract, sites other than the lung or upper aerodigestive tract, and the lung, respectively. Survival was statistically reduced when an MSPC developed; the 10-year actuarial survival estimates were 76.8% in patients who did not develop an MSPC and 43.7% in patients who developed an MSPC (p < .0001). Overall, 68.4% of patients who developed an MSPC (67/98) died of this disease. The 10-year actuarial survival estimates were 24% for lung MSPCs, 43.7% for non-lung, non-upper aerodigestive tract MSPCs, and 63.4% for upper aerodigestive tract MSPCs. PMID- 11913680 TI - Association of Epstein-Barr virus and nasopharyngeal carcinoma in Caucasian patients. AB - We evaluated the association of nasopharyngeal carcinoma (NPC) and Epstein-Barr virus (EBV) in Spanish patients, and studied the expression of EBV products (latent membrane protein-1 [LMP-1] and ZEBRA proteins) by NPC cells and its possible prognostic value. In situ hybridization (ISH) for EBV-encoded nonpolyadenylated RNAs (EBERs) and immunohistochemical expression of LMP-1 and ZEBRA proteins by immunohistochemistry were examined in formalin-fixed, paraffin embedded NPC specimens from 30 patients, and a survival analysis was done by the Kaplan-Meier method. We detected EBERs by ISH in 96.67% of the NPC cases, and detected expression of LMP-I in 43.33% of the NPC cases and expression of ZEBRA protein in 6.67% of the NPC cases. We conclude that ISH for expression of EBERs is an adequate method for detection of EBV in NPC. LMP-1 is not frequently expressed in NPC cells (43.33%). Most NPC cells carry a latent EBV infection. LMP 1 expression might have worsened the prognosis of NPC in our series. PMID- 11913681 TI - Median cleft of the lower lip: report of two new cases and review of the literature. AB - Median clefts of the lower lip and mandible are rare craniofacial clefts. Couronne in 1819 was the first to describe the condition. The midline cleft of the lower lip was classified by Tessier as a type 30 craniofacial cleft. Recently, the total number of the reported cases had increased to about 66 in the world literature. In addition, 2 more patients are presented here. The first case involves only a small notch in the vermilion; the deformity was treated by Z plasty. In the second case, a midline incomplete cleft of the lower lip, a sublingual abnormal frenulum, a complete cleft in the mandible, a bifid sternum, a presternal skin tag, and a ventricular septal defect in the heart were present. At operation, Z-plasty of the lingual frenulum released the normal-sized tongue. The lip cleft was corrected by a simple V excision followed by closure in 3 layers. The mandibular segments were "vitalized" with a bone rongeur and immobilized in the predetermined position with an interosseous stainless steel wire. Because the bone fusion was complete, the stainless steel wire was taken out after 3 months so that it would not prevent mandibular development. Our treatment methods and others are discussed. PMID- 11913682 TI - Topical estrogens combined with argon plasma coagulation in the management of epistaxis in hereditary hemorrhagic telangiectasia. AB - The aim of this study was to assess the value of topically applied estrogens in patients with hereditary hemorrhagic telangiectasia. Twenty-six patients with this disorder were treated with argon plasma coagulation and randomized into 2 groups: group A, which had postoperative application of estriol ointment (n = 14), and group B, which had postoperative application of dexpanthenol ointment (n = 12). Over a period of 12 months, the frequency and intensity of bleeding, the patient's satisfaction, and the success of the treatment were evaluated with a questionnaire. Before the operation, more than 90% of the patients in both groups complained of daily episodes of epistaxis. Twelve months after treatment, the frequency and intensity of bleeding had significantly decreased in group A as compared to group B. Of the patients in group A, 93% were satisfied with the treatment. Of the patients in group B, only 42% were satisfied with the treatment. In both groups, more than 90% of the patients were willing to undergo the same treatment again. The combined treatment approach with argon plasma coagulation and topical estriol enables us to significantly prolong the hemorrhage-free interval. PMID- 11913683 TI - Occurrence of familial Meniere's syndrome and migraine in Brasilia. AB - Since 1941, many families with several members affected by Meniere's syndrome have been described. "Migraine-like" headache was always mentioned. In 1997, we described a large family with Meniere's syndrome and migraine, both transmitted in an autosomal dominant mode. Recently, a gene that could be responsible for Meniere's symptoms has been identified. However, the incidence of familial Meniere's syndrome is still unknown. A. W. Morrison et al estimated this incidence as between 2.9% and 12%. During the years 1997 and 1998, we saw 8 new patients with typical Meniere's syndrome in our outpatient department. They were referred to Dr Messias, who did a careful search for other affected family members. Six of them had relatives with Meniere's syndrome and migraine transmitted in an autosomal dominant mode. The familial incidence of Meniere's syndrome and migraine in Brasilia seems to be higher than the figures of Morrison et al for the United Kingdom. The 6 families are described herein. PMID- 11913684 TI - Vascular occlusion in the endolymphatic sac in Meniere's disease. AB - In 2 patients with severe Meniere's disease (MD), there was histologic evidence of occlusion of the vein of the vestibular aqueduct (VVA). This finding coincided with total or partial occlusion of numerous small vessels around the endolymphatic sac (ES), flattening of epithelium, extensive perisaccular fibrosis, and signs of new bone formation. Ultrastructural analysis of the occluding material showed foci with dense connective tissue, calcification, lipid deposits, and layers of basement membrane, sometimes concentrically arranged. The exact nature of the occluding material was unknown. In another 2 MD patients, the VVA was not visualized, and the ES vessels showed no signs of occlusion. Seven controls with acoustic schwannoma or meningioma had normal vasculature. The presence of vascular impairment in the ES in MD patients indicated that altered hemodynamics may contribute to the pathogenesis of endolymphatic hydrops and MD. PMID- 11913685 TI - Malleus fixation: clinical and histopathologic findings. AB - The goals of this study were to review important clinical and histopathologic features of malleus fixation. Ten clinical cases and 10 histopathologic cases of malleus fixation were identified. For the clinical cases, preoperative clinical data, surgical findings, and preoperative and postoperative audiometric findings were reviewed. Ninety percent of the clinical cases achieved significant reduction of the air-bone gap after operation. Two thirds of these cases had an air-bone gap of 10 dB or less, and the remainder had air-bone gaps between 20 and 28 dB after operation. For the temporal bone cases, clinical data, histopathologic findings, and other otologic diagnoses were reviewed. Malleus fixation can be idiopathic or a result of trauma, chronic otitis media, or developmental anomalies. It is a cause of hearing loss that is likely to be more common than the number of diagnoses would indicate. Certain audiometric findings may lead one to suspect the diagnosis. The surgical approach used depends on the individual anatomy, and surgery is usually highly effective in improving hearing. PMID- 11913686 TI - Estimated locations of the narrowest portion of the eustachian tube lumen during closed and open states. PMID- 11913687 TI - Auditory consequences of recurrent acute purulent otitis media. AB - To investigate whether recurrent purulent otitis media results in permanent hearing loss, we studied 2 subgroups of children from a cohort, earlier prospectively followed from birth to the age of 3 years. One subgroup had recurrent acute otitis media (n = 12), and the other had no acute otitis media at all ("healthy" children; n = 21). At follow-up of these subgroups at the age of 10, no child had acute otitis media or secretory otitis media. There was no difference between the groups in hearing level thresholds at the frequencies 125 Hz to 8 kHz. However, in the children with recurrent acute otitis media, as compared with the controls, the hearing levels at high frequencies (8 to 16 kHz) and the acoustic middle ear reflex thresholds were elevated, the middle ear compliance was higher, and click-evoked otoacoustic emission response levels and middle ear pressures were lower. The results suggest that the middle ear mechanics of children with recurrent acute otitis media are affected, and also that their cochlear function might be disturbed. PMID- 11913688 TI - Expression of heat shock protein 70 in the cochlea in experimental autoimmune inner ear disease. AB - To explore whether the immune response of the inner ear could induce heat shock protein 70 (hsp70) in the guinea pig cochlea, we established a model of autoimmune inner ear disease (AIED) by systemically immunizing guinea pigs with homologous crude inner ear antigen (CIEAg). The immunized cochleas and normal control cochleas were examined for the expression of hsp70 by immunohistochemical analysis and in situ hybridization. It was found that 10 of 28 ears showed significant increases in hearing thresholds after the animals had been sensitized with CIEAg, and 67.9% of the ears from the immunized animals had modiolar vasculitis. The hsp70-like proteins were basically expressed in the normal cochleas, but dramatically increased expressions of hsp70 (perhaps also including hsp70-like molecules) and its messenger RNA were observed in the cochleas of the AIED models. The results suggest that the immune response of the inner ear induced the expression of hsp70 in the guinea pig cochleas. PMID- 11913689 TI - Imaging case study of the month. Pediatric virtual bronchoscopy. PMID- 11913690 TI - A benign sessile endobronchial leiomyoma in a 2-year-old girl with human immunodeficiency virus (HIV). PMID- 11913691 TI - ADSA Foundation Scholar Award. Formation and physical properties of milk protein gels. AB - Gelation of milk proteins is the crucial first step in both cheese and yogurt manufacture. Several types of milk gels are discussed, with an emphasis on recent developments in our understanding of how these gels are formed and some of their key physical properties. Areas discussed include the latest dual-binding model for casein micelles; some recent developments in rennet-induced gelation; review of the methods that have been used to monitor milk coagulation; and a discussion of some of the possible causes for the wheying-off defect in yogurts. Casein micelles are the primary building blocks of casein-based gels; however, controversy about its structure continues. The latest model proposed for the formation of casein micelles is the dual-binding model proposed by Horne, 1998, which suggests that casein micelles are formed as a result of two binding mechanisms, namely hydrophobic attraction and colloidal calcium phosphate (CCP) bridging. Most previous models for the casein micelle have treated milk gelation from the viewpoint of simple particle destabilization and aggregation, but they have not been able to explain several unusual rheological properties of milk gels. Although there have been many techniques used to monitor the milk gelation process over the past few decades, only a few appear attractive as possible in vat coagulation sensors. Another important aspect of milk gels is the defect in yogurts called wheying-off, which is the appearance of whey on the gel surface. The factors responsible for its occurrence are still unclear, but they have been investigated in model acid gel systems. PMID- 11913693 TI - Vitamin A degradation and light-oxidized flavor defects in milk. AB - To determine the effects of light exposure on vitamin A degradation and on light oxidized flavor development, samples of whole, reduced fat, and nonfat milk were exposed to fluorescent light (either 1000 or 2000 lx) at time intervals of 2, 4, 8, or 16 h. Measurable vitamin A losses occurred at 2, 4, and 16 h at 2000 lx for nonfat, reduced fat and whole milk, respectively. Moderate light-oxidized flavors were detected after 4 h of light exposure (2000 lx) in the whole and reduced fat milk and after 8 h in nonfat milk. The different types of milk show a significant difference in relative flavor scores. By 16 h at 2000 lx, relative light-oxidized flavor development was lower in nonfat milk than in whole or reduced fat milk. The presence of milk fat appears to protect against vitamin A degradation in fluid products, but adversely affects the flavor quality of milk after exposure to light. In summary, these findings demonstrate that even a brief, moderate light exposure (2 h; 2000 lx) can reduce the nutritional value and flavor quality of fluid milk products. PMID- 11913692 TI - The composition of bovine milk lipids: January 1995 to December 2000. AB - Data from recent publications on bovine milk lipids are presented and discussed. This includes extraction of lipids, triacylglycerols, phospholipids, other complex lipids, sterols, isoflavones, and fatty acids. Improved gas-liquid and high performance liquid chromatography were used. Data on the trans and cis isomers of fatty acid and of conjugated linoleic acids are given, and the analyses are described. Papers about the lipids in milks and dairy products from the United States are few; where with the exception of trans-fatty acid isomers and conjugated linoleic acids, almost no research has been reported. PMID- 11913694 TI - The impact of wood ice cream sticks' origin on the aroma of exposed ice cream mixes. AB - The effect of volatile compounds in white birch sticks obtained from four different geographical locations on the aroma of ice cream mix was investigated. Sensory evaluation, (specifically, a series of warmed-up paired comparisons) was conducted on stick-exposed ice cream mixes to determine whether aroma differences in those mixes could be detected. Batches of ice cream mix were exposed to the sticks and aged for 6 d at 4 degrees C and then assessed by the panelists by pairwise comparison. Findings suggest that differences in aroma of mixes that have been exposed to white birch sticks from four different geographical origins can be distinguished perceptually. PMID- 11913695 TI - Effects of pair versus individual housing on the behavior and performance of dairy calves. AB - This study compared the health, performance, and behavior of individually and pair-housed calves fed milk ad libitum by artificial teats. Calves were separated from their dams within 24 h of birth and assigned to housing in either a single pen (10 calves) or a group pen (10 pairs of pair-housed calves). Calves were gradually weaned at approximately 5 wk of age and remained on the experiment until wk 8. Behavior was video recorded during wk 2 to 8. Before and after weaning, calves gained weight steadily with no differences between treatments. During the week of weaning, pair-housed calves continued to gain weight normally, but the individually housed calves experienced a growth check. There were no differences between groups in the amounts of milk, starter, or hay consumed, or in the incidence of scouring. There were also no differences in the amount of time spent self-grooming, sucking on the teat, or lying down. However, pair housed calves spent more time standing inactive, more time moving, and less time with their head out of the pen than individually housed animals. Paired calves spent approximately 2% of the day in social contact, and the incidences of agonistic behavior and cross-sucking were very low. These results indicate that housing dairy calves in pairs allows benefits such as increased space for movement and social opportunities with no disadvantages in health and weight gains. PMID- 11913697 TI - Influence of estrus on somatic cell count in dairy goats. AB - The effect of estrus on the somatic cell count (SCC) of goat's milk was examined by inducing estrus in 24 of 48 seasonally anestrus, lactating dairy goats. Goats were blocked by infection status and ranked on SCC from three preceding herd tests and randomly allocated (within block) to the following three treatment groups: a) "Short," in which an intravaginal progesterone-releasing device was inserted for 12 d plus equine chorionic gonadotropin and dinoprost tromethamine 2 d before device removal (n = 12), b) "Long," in which an intravaginal progesterone-releasing was inserted for 17 d plus equine chorionic gonadotropin on the day of device removal (n = 12), or c) "Control," in which the goats were left as untreated controls (n = 24). Bacteriological status of each gland of each goat was determined before and after synchronization (d -23 and +13) and SCC and milk volumes were determined on d -2, 0, 1, 2, 3, 4, 14, and 25, where d 0 was the day of intravaginal device removal. Goats in the Short group were in estrus before those of the Long group, who were, in turn, in estrus before the Control group. The log10 and log10 absolute SCC (SCC cells/ml x volume) were higher in the Short than in the Control group on d 1, 2, 3, and 4, whereas those of the Long group were higher than those of the Control group on d 2 and 4. These data indicate that estrus resulted in an increase in SCC, and that the increase in SCC was independent of the decline in milk volume at estrus. PMID- 11913696 TI - Genetic manipulation of the IGF-I axis to regulate mammary gland development and function. AB - Insulin-like growth factor I (IGF-I) is known to regulate mammary gland development. This regulation occurs through effects on both cell cycle progression and apoptosis. Our laboratory has studied the IGF-I-dependent regulation of these processes by using transgenic and knockout mouse models that exhibit alterations in the IGF-I axis. Our studies of transgenic mice that overexpress IGF-I during pregnancy and lactation have demonstrated that this growth factor slows the apoptotic loss of mammary epithelial cells during the declining phase of lactation but has minimal effects during early lactation on milk composition or lactational capacity. In contrast, our analysis of early developmental processes in mammary tissue from mice carrying a targeted mutation in the IGF-I receptor gene suggests that IGF-dependent stimulation of cell cycle progression is more important to early mammary gland development than potential anti-apoptotic effects. With both models, the effects of perturbing the IGF-I axis are dependent on the physiological state of the animal. The diminished ductal development that occurs in response to loss of the IGF-I receptor is dramatically restored during pregnancy, whereas the ability of overexpressed IGF I to protect mammary cells from apoptosis does not occur if the mammary gland is induced to undergo forced involution. Data from our laboratory on the expression of IGF-signaling molecules in the mammary gland suggest that this effect of physiological context may be related to the expression of members of the insulin receptor substrate family. PMID- 11913698 TI - Timing of onset of somatotropin supplementation on reproductive performance in dairy cows. AB - The objective of this field trial was to determine whether delaying the start of recombinant bovine somatotropin (rbST) supplementation from 9 to 10 wk postpartum to 17 to 18 wk postpartum would improve reproductive performance in lactating dairy cows. Cows from nine herds (n = 798 cows; 766 Holsteins, 32 Jerseys) were assigned at random to receive rbST supplementation at 14-d intervals beginning during wk 9 to 10 (n = 399) or wk 17 to 18 (n = 399) after calving. Effects of herd, season of calving, parity, and onset of rbST supplementation (9 to 10 wk vs. 17 to 18 wk) on days to first service and days open were determined. In primiparous but not multiparous cows, there tended to be fewer days to first service and fewer days open when onset of rbST supplementation was delayed. Percentages of cows pregnant at 150, 200, and 250 d postpartum were also examined. Time of onset of rbST did not affect percentages of multiparous cows pregnant at 150, 200, and 250 d postpartum. However, there appeared to be a slight tendency for percentages of pregnant primiparous cows to be greater at 200 and 250 d postpartum for those receiving rbST supplementation beginning at 17 to 18 wk compared to those receiving rbST starting at 9 to 10 wk. In conclusion, delaying the start of rbST supplementation to wk 17 to 18 postpartum had no beneficial effect on reproductive performance of multiparous cows but tended to improve some measures of reproductive performance in primiparous cows. PMID- 11913699 TI - Effect of season and exposure to heat stress on oocyte competence in Holstein cows. AB - Two experiments were conducted to evaluate seasonal variation in oocyte competence in Holstein cows and to test whether oocyte quality in summer is affected by the magnitude of heat stress. In the first experiment, ovaries of Holstein cows were collected from a slaughterhouse and used to harvest oocytes over 1 yr (n = 18 replicates). After in vitro maturation, fertilization, and culture, proportions of oocytes and cleaved embryos that developed to blastocysts by d 8 were lower in the warm season compared with the cool season. In the second experiment, nonlactating Holstein cows were housed in one of the following three environments for 42 d before slaughter: heat stressed (housed with shade cloth in summer; n = 14); cooled (housed in a free-stall barn with foggers and fans in summer; n = 14); and winter (housed similar to the heat-stressed group; n = 12). Cows were slaughtered at d 18 to 19 of the estrous cycle. Oocytes from the two largest follicles per cow were aspirated and cultured individually. Ovaries were then dissected to collect additional oocytes that were processed in a group for in vitro maturation, fertilization, and culture. Cleavage rates were similar among treatments, but none of the individually cultured oocytes developed to blastocysts. For other oocytes cultured in groups, proportions of oocytes and cleaved embryos that developed to blastocysts by d 8 were lower in summer than winter with no difference between the heat-stressed and the cooled treatment groups. Summer depression in oocyte quality in Holstein cows was evident, but cooling cows for 42 d did not alleviate that seasonal effect. PMID- 11913700 TI - Prepartum monensin for the reduction of energy associated disease in postpartum dairy cows. AB - A total of 1317 Holstein cows from 45 farms in the Canadian provinces of Quebec, Prince Edward Island (PEI) and Ontario were enrolled in a randomized trial during 1998 and 1999 to further confirm the efficacy of a monensin controlled release capsule in preventing periparturient disease in lactating dairy cows. Cows were randomized on the farms to receive either a monensin controlled release capsule (CRC) 2 to 4 wk before expected calving or to serve as negative controls. Health data were collected for 90 d postcalving and were analyzed with logistic regression accounting for the intraherd correlation with generalized estimating equations. Monensin CRC significantly reduced the incidence of both clinical ketosis and abomasal displacement post-calving. There was a numerical but nonsignificant decrease in the incidence of retained placenta in cows receiving a monensin CRC. A pooled analysis of two separate but similar studies (conducted in 1995 and 1998) demonstrated a strengthened association between monensin CRC administration precalving and reduced periparturient disease. A 40% reduction in both abomasal displacement and clinical ketosis was observed with precalving administration of a monensin CRC. In addition, the larger dataset highlighted a trend for a 25% reduction in the incidence of retained placenta in monensin treated cows. Improved energy metabolism as a result of monensin treatment is likely the mechanism for the reduction in incidence of all three of these diseases. Thus the term "energy associated disease" was created to assess the combined impact of the precalving monensin treatment on the incidence of retained placenta, displaced abomasum, and clinical ketosis. The monensin controlled release capsule reduced the incidence of energy associated disease by 30%. PMID- 11913701 TI - Paratuberculosis in dairy cattle: variation of the antibody response in offspring attributable to the dam. AB - The objective of this study was to examine transmission of paratuberculosis in dairy cattle attributable to the dam. Milk samples were collected from 8131 cows in 110 Danish dairy herds. The level of antibodies to Mycobacterium avium subspecies paratuberculosis was determined by use of an ELISA. Information on dam and sire was obtained from the Danish Cattle database. The following two data sets were analyzed: Data set A contained all cows < or = 400 days in milk (n = 7410); data set B contained 1056 dam-daughter pairs present simultaneously in herds at the day of sampling. Cows > 400 days in milk were excluded. Linear mixed models were used to obtain variance components for the effect of sire in data set A and the effect of sire and dam-daughter pairs in data set B. Models for both data sets A and B included information previously shown to confound antibody level and information of the relative prevalence of paratuberculosis in the herd. In data set A, the effect explained by sire was 1.9%, whereas it was 6.3% in data set B. The effect from dam-daughter pairs was 7.7%. Those effects were all significant. It was concluded that the parental contribution was significant, and both heritability of susceptibility and vertical transmission should be considered in any control programs on paratuberculosis in dairy cattle. PMID- 11913702 TI - Effects of spray-dried animal plasma in milk replacers or additives containing serum and oligosaccharides on growth and health of calves. AB - The effects of spray-dried animal plasma in milk replacer without or with the addition of additives containing fructooligosaccharides and spray-dried serum on health, growth, and intake of Holstein calves was measured in two 56-d experiments. In experiment 1, 120 calves were fed milk replacer containing 0 or 20% of crude protein as spray-dried bovine plasma for 42 d and 30 to 60 g/d of additives containing whey protein concentrate or bovine serum for the first 15 d. Commercial calf starter was available from d 29, and water was available at all times. In experiment 2, 120 calves were fed milk replacer containing 0 or 16% of crude protein as spray-dried bovine plasma with 0 or 30 to 60 g/d of additive containing bovine serum for the first 15 d. Additive containing bovine serum also contained fructooligosaccharides, whey, and vitamin/mineral premix. In experiment 1, calves fed additive containing bovine serum tended to have fewer days with diarrhea, lower use of electrolytes, and improved BW gain from d 29 to 56. The addition of spray-dried bovine plasma to milk replacer did not influence any parameter measured. In experiment 2, calves fed additive containing bovine serum or milk replacer containing spray-dried bovine plasma had lower mortality (4.4 vs. 20%) and tended to have improved fecal scores and fewer days with scours. Antibiotic use was lower when calves were fed the additive. Indices of enteric health (incidence of scours and treatment with antibiotics and electrolytes) were improved when plasma was added to milk replacer throughout the milk feeding period or as an additive during the first 15 d of the milk feeding period, when calves were most susceptible to enteric pathogens. The addition of spray-dried animal plasma to milk replacer or the addition of additive containing spray-dried bovine serum and oligosaccharides may be a useful adjunct to animal management during periods of stress. PMID- 11913703 TI - Evaluation of an indwelling ruminal probe methodology and effect of grain level on diurnal pH variation in dairy cattle. AB - To evaluate a ruminal probe for recording diurnal variation in rumen pH, we fitted three ruminally fistulated cows with probes affixed to the inside of the cannula. Probes were connected to a data logger and readings were recorded hourly. The experiment was a Latin square design with cows fed three different diets: 50, 60, and 70% grain as a total mixed rations once daily. Feed offerings and refusals were recorded daily. The experimental period was 21 d. The first 6 d were for adaptation, followed by 5-d rotations through each of the diets. Daily probe readings were recorded at 4-, 6-, 8-, 12-, and 24-h intervals. At each interval, readings were recorded (precleaned), a sample of rumen fluid was taken, and pH was measured in the laboratory. As probes were removed from the rumen, probe ends were cleaned with 0.1 N HCl and reinserted into the rumen, and a reading was recorded (postcleaned). No protective pH probe shields were used in this experiment. There were no differences between pre- and postcleaned pH readings across cows for all diets. Mean time under pH 5.0, 5.5, and 6.0 were 0.2, 2, and 7.2 h, respectively. Diet affected length of time under a certain pH, only for readings under pH 6. Diurnal pH profiles were monophasic in nature. The degree of acidity increased after feeding and duration of nadir increased with increasing grain in the diet. Daily DMI increased but was highly variable within the first week after switching to the next higher grain level. These results indicate the use of an indwelling ruminal probe without a protective shield, cleaned, and calibrated daily can accurately measure diurnal variation of ruminal pH. In addition, transition to higher grain levels in the diet increases pH, duration of pH nadir, and daily DMI fluctuation. PMID- 11913704 TI - Corn silage management: effects of maturity, inoculation, and mechanical processing on pack density and aerobic stability. AB - Three experiments were conducted to evaluate the effects of inoculation, maturity, and mechanical processing of corn silage on aerobic stability and pack density. Corn silage was stored in 20-L mini silos for the three aerobic stability experiments. Corn silage was stored in 80-L mini silos for the three pack-density experiments. The wet pack density of corn silage tended to decrease as maturity advanced in all of the pack-density experiments, and processed corn silage had a greater wet pack density compared with unprocessed corn silage in two of the three 20-L mini silo experiments. Aerobic stability, measured as the number of hours to reach 1.7 degrees C above ambient, was greater for processed corn silage in two of the three 20-L mini silo experiments, and was greater for inoculated corn silage across the three 20-L mini silo experiments. Inoculation of corn silage with lactic acid producing bacteria tended to improve aerobic stability of corn silage more than maturity and mechanical processing. PMID- 11913705 TI - Genetic parameters of dairy character, protein yield, clinical mastitis, and other diseases in the Danish Holstein cattle. AB - The primary aim of this study was to estimate genetic correlations between dairy character, protein yield, clinical mastitis, and other diseases. Data consisted of first lactation records of Danish Holstein cows calving from 1990 to 1999. After editing, the data included records on 934,639 cows, of which 101,853 were assessed for dairy character, 472,421 for diseases, and 834,993 for protein yield. The disease traits were defined as binary traits in the period from 10 d before to 50 d after calving for clinical mastitis, and from 10 d before to 100 d after calving for diseases other than mastitis. Data were analyzed with a linear sire model using the method of AI-REML. Heritabilities were estimated to be 0.265 for protein yield, 0.261 for dairy character, 0.035 for clinical mastitis, and 0.020 for diseases other than mastitis. Estimates of genetic correlations between protein yield and dairy character, protein yield and clinical mastitis, and protein yield and diseases other than mastitis were 0.38, 0.33, and 0.14. Between the two disease traits, the genetic correlation was 0.24. The genetic correlation between dairy character and clinical mastitis was 0.24. Between dairy character and diseases other than mastitis the genetic correlation was 0.41. Thus, cows with high score for dairy character were more prone to diseases. The genetic correlation between dairy character and the disease traits, when both traits were adjusted for protein yield, was 0.13 for clinical mastitis and 0.39 for diseases other than mastitis. These findings suggest that, dairy character should be given a negative rather than a positive weight in the breeding goal. PMID- 11913706 TI - Large batch freezing of bull semen: effect of time of freezing and fructose on fertility. AB - Large-scale batch freezing of bull semen should be done in a processing schedule that yields the highest fertility and when it can be fitted efficiently into the work schedule. Conflicting reports have appeared on survival and fertility of bull sperm frozen within a few hours of semen collection or on the next day. To study this problem, a factorially arranged experiment with semen from 10 bulls was conducted, comparing whole milk-glycerol semen extender with and without fructose, and semen frozen in 0.5-ml straws after 4 versus 18 h of equilibration at 5 degrees C. Both fructose and 18 h of equilibration resulted in a small but significant improvement in freeze-thaw survival of sperm. A field trial followed with replicated semen collections from nine bulls processed in a whole milk glycerol control extender frozen after 4 h of equilibration versus the addition of 1.25% (wt/vol) fructose to whole milk glycerol divided to freeze sperm after 4 and 28 h of equilibration. Semen from these bulls was used to inseminate 14,775 first-service cows. The 56-d nonreturn rates obtained for these three treatments were 74.7, 74.3, and 73.9%, respectively. As there was no difference in fertility, it would appear that programs to freeze sperm in whole milk extenders the same day of collection or the day after semen collection should yield equivalent results. PMID- 11913707 TI - Serum leptin levels in different types of hypertension during pregnancy. AB - We determined the serum levels of leptin in 96 pregnant women with body mass index between 20 to 30, 30 normal (NP), 26 with mild preeclampsia (MPE), 27 with severe preeclampsia (SPE), 6 with chronic hypertension plus preeclampsia (CHT+PE) and 7 with chronic hypertension (CHT). A significant (p < 0.01) decrease in leptin levels was observed in the SPE group when compared with the NP group. On the contrary, significant (p < 0.05) increases were observed in the CHT and CHT+PE groups when compared with the NP group. Leptin levels were significantly higher in the MPE (p < 0.001), CHT (p < 0.01) and CHT+PE (p < 0.5) groups when compared with the SPE. No significant differences were observed in the CHT group when compared with CHT+PE. Moreover, a positive correlation was encountered (r = 0.6, p < 0.001) between platelet number and leptin levels for all the patients with preeclampsia. These results suggest that leptin levels may be useful metabolic parameter in different types of hypertension during pregnancy. PMID- 11913708 TI - Enhanced expression of microsomal epoxide hydrolase and glutathione S-transferase by imidazole correlates with the radioprotective effect. AB - Previous studies have shown that induction of microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) by oltipraz correlated with the radioprotective effect. The present study was designed to investigate the expression of the antioxidant enzymes and the radioprotective effect by imidazole (IM). Northern blot analysis revealed that IM increased the mEH and GST mRNA levels in the rat liver in a dose-dependent manner. Rats irradiated with 3 Gy of gamma-rays in combination with IM showed enhanced increases in mEH and rGSTA2 mRNAs, as compared to either IM or irradiation alone. IM prevented elevations in the hepatic GSH content by gamma-irradiation. In contrast to IM, cysteine blocked radiation-inducible increases in the mRNAs with no suppression of the GSH content. The radioprotective effect by IM was greater than that by cysteine, as assessed by the 30-day survival rate of mice (i.e. 80% and 69%, respectively, vs. 48% in control). These results demonstrated that IM enhanced radiation-inducible mEH and GST expression with prevention of the increase in GSH content, which correlated with the radioprotective effect, and that the mechanistic basis of radioprotection by IM differed from that by cysteine. PMID- 11913709 TI - Factors influencing the protein binding of IQO4, a new isoquinolinedione derivative. AB - Various factors most likely to influence the plasma protein binding of IQO4, a new isoquinolinedione derivative, to 4% human serum albumin (HSA) were evaluated using an equilibrium dialysis technique at an initial IQO4 concentration of 5 microg/ml. It took approximately 12 h incubation to reach an equilibrium between 4% HSA and isotonic Sorensen phosphate buffer at pH 7.4 containing 3% dextran ('the buffer') using a Spectra/Por 2 membrane (molecular weight cut-off, 12000 14000) in a water-bath shaker kept at 37 degrees C and at a rate of 50 oscillations per min. IQO4 was stable both in 4% HSA and in 'the buffer' for up to 24 h incubation at 37 degrees C. The binding of IQO4 was constant (89.9 +/- 1.40%, mean +/- standard deviation) at IQO4 concentrations ranging from 1 to 100 microg/ml. However, the extent of binding was dependent on HSA concentrations. The values were 32.5, 62.0, 79.1, 84.9, 90.9, 91.2, and 91.7% at HSA concentrations of 0.5, 1, 2, 3, 4, 5, and 6%, respectively; on incubation temperature, 96.7, 93.8, and 91.0% when incubated at 4, 22, and 37 degrees C, respectively; and on the buffer pHs, 84.4, 87.2, 88.2, 90.9, and 92.3% for the buffer pHs of 5.8, 6.4, 7, 7.4, and 8, respectively. The free fraction of IQO4 increased with the addition of sulfisoxazole (0-300 microg/ml), and salicylic acid (0-300 microg/ml). The protein binding of IQO4 was independent of the quantity of alpha-1-acid glycoprotein (up to 0.32%), chloride ion (up to 0.546%) and heparin (up to 40 units/ml). PMID- 11913710 TI - Soluble Fas and soluble Fas L levels in patients with acute pancreatitis. AB - The FasL-Fas system is an apoptosis induction system and plays an important role in homeostasis and biophylaxis. The present study was conducted to investigate soluble Fas (sFas), soluble Fas ligand (sFasL), and tumor necrosis factor alpha (TNF-alpha) in patients with acute pancreatitis. As acute pancreatitis became severe, the levels of sFas in the serum increased significantly, while those of sFasL decreased significantly. A significant inverse correlation was observed between the serum levels of sFas and those of sFasL. Also, a significant positive correlation was observed between the levels of TNF-alpha and sFas. A greater increase in serum sFas and decrease in serum sFasL levels was observed in patients with complicating multiple organ dysfunction syndrome (MODS) than in those without it. The results of the study suggest that the pathological aggravation of acute pancreatitis could be related to changes in the Fas-FasL system. PMID- 11913711 TI - Pharmacological aspects of ipecac syrup (TJN-119)-induced emesis in ferrets. AB - In order to elucidate the precise mechanism of ipecac syrup (TJN-119) on the occurrence of vomiting, we examined the effects of ipecac syrup on the abdominal afferent nerve activity as well as on the 5-HT levels of the ileum and area postrema in ferrets. Oral administration of TJN-119 (0.5 mg/kg) produced a significant increase in afferent abdominal vagus nerve activity which lasted approximately 1 hour. The maximum response induced by TJN-119 was estimated to be 219 +/- 18% of the pre-injection level. Cephaeline or emetine, the main alkaloids of ipecac syrup, also demonstrated similar effects on afferent vagus nerve activity. TJN-119 increased the 5-HT content in the ileum but not in the area postrema. These observations illustrate possible mechanisms that may act at peripheral sites. It was recently reported that TJN-119 has a high affinity to 5 HT4 receptors (Hasegawa et al., unpublished data). These results suggest that 5 HT4 receptors may be involved in the emetic action of TJN-119. PMID- 11913712 TI - Metabolic changes in DOCA-salt hypertensive rats. AB - In a previous report, we observed an altered proportion of fiber types and a reduction of capillary per fiber ratio in extensor digitorus longus (EDL) and soleus (SOL) muscles of deoxicorticosterone acetate (DOCA)-salt hypertensive rats when compared with controls. The aim of the present study was to ascertain various carbohydrate and lipid enzyme activities and substrates that may be involved in the morphological changes reported. In the SOL muscle of hypertensive rats, glucose, glycogen and triglycerides (TG) levels were increased, citrate synthase (CS) and beta-hydroxy-acyl-CoA dehydrogenase (HAD) activities were reduced, while hexokinase (HK) and lipoprotein lipase (LPL), LPL mass, lactate and free fatty acids (FFA) levels were unchanged. In EDL muscles of hypertensive rats, glycogen levels and LPL mass were higher than in controls, while CS, HAD, HK, and LPL activities and glucose, lactate, FFA and TG levels were unmodified. Serum levels of insulin, TG, cholesterol and FFA were increased while glucose levels were decreased and high-density lipoprotein-cholesterol levels were similar in hypertensive rats when compared with controls. In conclusion, hypertensive rats showed increased glycogen in both EDL and SOL muscles, with hyperinsulinemia and reduced glycemia. Hyperinsulinemia might have been a compensatory response to insulin resistance. The oxidative capacity of SOL muscle was reduced indicating that glucose uptake was conduced via non-oxidative metabolism. TG, FFA and cholesterol were increased in serum and TG in SOL muscle. PMID- 11913713 TI - Undernutrition during early lactation period induces metabolic imprinting leading to glucose homeostasis alteration in aged rats. AB - This study examines the effects of early postnatal undernutrition on the glucose homeostasis of rats at one year of age, comparing the effects of a free protein diet (FPD) and a normal diet containing 25% of protein (NPD) supplied during the first 10 days of lactation. The insulin secretion and the insulin sensitivity, using the glucose clamp technique, were studied in these rats. The analysis of the integrated area of insulin secreted by isolated islets stimulated with 16.7 mM glucose was reduced in the FPD group when compared with the NPD (FPD = 5.07 +/ 1.6 ng/ml/50 min.; NPD = 35.8 +/- 12 ng/ml/50 min., p < or = 0.001). Using the glucose clamp technique the plasma glucose concentration was raised by continuous glucose stimulation with 10 mg/Kg(-1) x min(-1). After 30 minutes the NPD displayed a lower level of plasma glucose concentration (FPD = 220.8 +/- 8 mg/dl.; NPD = 185 +/- 3 mg/dl., p < or = 0.01). Afterwards, the hyperglycemia of the NPD increased and in both groups was, subsequently, similarly maintained and, after 90 minutes of continuously glucose infusion, there was no difference between the groups (FPD = 191.6 +/- 8 mg/dl.; NPD = 189.3 +/- 17 mg/dl). In order to test the peripheral sensitivity to glucose, insulin 1.67 mU x Kg(-1) min(-1) was administered together with glucose 10 mg x kg(-1) x min(-1) (50 minutes after the beginning of the clamping). The glycemia after the insulin administration compared to glycemia 90 minutes of FPD was significantly reduced and the NPD maintained the same glycemic level ( FPD from 220.7 +/- 8 mg/dl to 170.6 +/- 5 mg/dl, p < or = 0.001; NPD from 195.3 +/- 10 mg/dl to 185.2 +/- 6 mg/dl.). Also, after the insulin administration the plasmatic insulin was raised but after 90 minutes the FPD group displayed a lower insulin concentration when compared to the same point of time for the NPD group (FPD = 0.8 +/- 0.01 ng/ml; NPD = 1.8 +/- 0.03 ng/ml., p < or = 0.01). The data suggest that undernutrition during early postnatal may cause a metabolic imprinting which leads to a decreasing action of the insulin secretory apparatus and increased insulin sensitivity as an adaptive response. PMID- 11913714 TI - Effect of exercise training on ANP receptors. AB - To clarify whether exercise-training affects ANP function, we trained male Wistar rats by treadmill running for nine weeks and measured ANP receptor number and affinity in the kidney, lung and adrenal. We also measured guanylate cyclase activity, by which second messenger cGMP accumulates. The number of adrenal ANP receptor significantly increased after exercise-training. There was no significant difference of affinity for all the organs examined between the training group and the control group. Guanylate cyclase activity tended to decrease in the kidney in the training group. A significant difference was found when the samples were stimulated by C-type natriuretic peptide (CNP). There was no significant difference in guanylate cyclase activity in the lung and adrenal. These findings are consistent with the exercise-induced hypervolemia, but not with the anti-hypertensive role of exercise-training. PMID- 11913715 TI - Acetaminophen-induced immunosuppression associated with hepatotoxicity in mice. AB - We investigated the influence of acetaminophen (APAP), an analgesic and hepatotoxic agent, on the immune system in mice. The activity of serum glutamic pyruvic transaminase was markedly increased by about 200 fold compared to that of the vehicle control following intraperitoneal injection of 400 mg/kg of APAP. In vivo antibody-producing responses to SRBC was significantly inhibited by APAP in a dose-dependent manner, while in vivo T cell-independent antibody-producing responses to TNP-Ficoll was not inhibited. The addition of thymocytes from APAP treated mice suppressed the response to SRBC in vitro. Thymocyte blastogenesis following mitogenic stimulation with concanavalin A was also inhibited by injection of APAP. The delayed-type hypersensitivity response and mixed lymphocyte reaction, which are used to evaluate cell-mediated immunity, were also significantly reduced after treatment with APAP. These results indicate that APAP suppresses the humoral and cell-mediated immune responses at a dose that causes liver injury. PMID- 11913716 TI - Do the CK18 related proteins change in general in epithelial cancers? AB - The modulation of cytokeratin 18 during tumor transformation in hepatoma had been previously recognized through a series of biochemical and immunological approaches. Expression of cytokeratin 18 in transitional cell carcinoma comparing with hepatoma was investigated using the hepatoma transformation model. CK18 related molecules were found. In the present study, we design various epithelial cancers with the same model. CK18 related molecules were all evident. Therefore, we suggest that CK18 related proteins would play an important role in tumorigenesis of epithelial cancers. PMID- 11913717 TI - Effects of neostigmine on the pharmacokinetics of intravenous parathion in rats. AB - It was reported that the area under the plasma concentration-time curve from time zero to time infinity (AUC) of parathion was significantly smaller and the time averaged total body clearance (CL) of parathion was significantly faster after intravenous administration of parathion to rats pretreated with dexamethasone than those in control rats. This was supported by significantly faster intrinsic clearance of parathion to form paraoxon in hepatic microsomal fraction of rats pretreated with dexamethasone. The above data suggested that parathion was metabolized to paraoxon by dexamethasone-inducible hepatic cytochrome P450 (CYP) 3A in rats. The purpose of this study is to explain the protective effects of neostigmine against paraoxon toxicity by suppressing CYP3A and hence decreasing formation of toxic metabolite, paraoxon by neostigmine. The pharmacokinetic changes of parathion and its active metabolite, paraoxon, were investigated after intravenous administration of parathion, 3 mg/kg, to control Sprague-Dawley rats and the rats pretreated with neostigmine (200 microg/kg, intraperitoneal injection 30 min before parathion administration). After 1-min intravenous infusion of parathion to rats pretreated with neostigmine, the AUC of parathion (65.1 versus 74.3 microg min/ml) was significantly greater and the CL of parathion (45.1 versus 40.4 ml/min/kg) was significantly slower than those in control rats. Based on in vitro hepatic microsomal studies, neostigmine inhibited significantly the erythromycin N-demethylase activity (1.03 versus 0.871 nmol/mg protein/min), mainly mediated by hepatic cytochrome P450 3A in rats. The above data suggested that the formation of paraoxon was inhibited in rats pretreated with neostigmine by inhibiting CYP3A. PMID- 11913719 TI - Significance of the human genome sequence to drug discovery. PMID- 11913720 TI - The C/At is out of the bag: a gene for mental illness. PMID- 11913718 TI - Pharmacokinetics, stability, and blood partition of DA-8159, a new phosphodiesterase V inhibitor. AB - The pharmacokinetics of DA-8159, a new phosphodiesterase V inhibitor, after 1 min intravenous, 30 mg/kg, and oral, 30 mg/kg, administration of the drug to rats, the stability of DA-8159 in various pH solutions ranging from 1 to 13, and human and rat plasma and urine, and the blood partition of DA-8159 between plasma and blood cells of rabbit were evaluated. After intravenous administration, DA-8159 was eliminated fast with the mean total body clearance of 126 ml/min/kg, and was almost completely metabolized in rats; 5.98% of intravenous dose of DA-8159 were excreted unchanged in 24-hr urine. The extent of absolute oral bioavailibility of DA-8159 was approximately 25%. The apparent volume of distribution at steady state was considerably large, 15048 ml/kg, suggesting that DA-8159 has a good affinity to rat tissues. DA-8159 was relatively stable in various pH solutions, and human and rat plasma and urine for up to 48 h incubation in a water-bath shaker kept at 37 degrees C and at a rate of 50 oscillations per min. DA-8159 reached equilibrium fast (within 30 sec mixing manually) between plasma and blood cells of rabbit blood and the plasma-to-blood cell concentration ratios were independent of initial blood concentrations of DA-8159, 1, 5, and 10 microg/ml, when the rabbit whole blood was incubated for up to 120 min; the ratios were in the range of 0.662-0.812. There was no in vitro 'blood storage effect' in the plasma concentration of DA-8159. PMID- 11913721 TI - Large-scale proteomics and its future impact on medicine. PMID- 11913722 TI - Pharmacogenomics, ethnicity, and susceptibility genes. PMID- 11913723 TI - Pharmacogenetics and the future of medical practice: conceptual considerations. PMID- 11913724 TI - The pharmacogenetics of asthma: a candidate gene approach. PMID- 11913725 TI - Site-specific molecular design and its relevance to pharmacogenomics and chemical biology. AB - The emergence of the new discipline of pharmacogenomics reflects the growing convergence of chemical and genomic space. The massive information-driven growth in both computational chemistry and structural biology is leading to unprecedented opportunities in both chemical and biological design. In this paper we relate current opinion in structural biology to recent developments in computational drug design. Sequence information now permits protein structure prediction and, together with experimental protein structure determination, a complete database of ligand-binding sites and protein-protein interactions can be assembled. When aligned with site exploration and virtual screening, this information provides a foundation for structure-based pharmacogenomics. In association with chemical genomics, structure-based design will allow major new insights into a compound's biological and pharmaceutical properties. PMID- 11913726 TI - The use of animal models in expression pharmacogenomic analyses. AB - Expression pharmacogenomics applies genome/proteome scale differential expression technologies to both in vivo and in vitro models of drug response to identify candidate markers correlative with and predictive of drug toxicity and efficacy. It is anticipated to streamline drug development by triaging towards lead compounds and clinical candidates that maximize efficacy while minimizing safety risks. As the majority of expression pharmacogenomics will be performed on preclinical therapeutic candidates, compatibility with favored preclinical animal model systems will be essential. This review will address expression pharmacogenomics in the context of those animal model systems commonly used for pharmacokinetic, pharmacodynamic and toxicologic analyses. Specific discussions will cover: (A) relative robustness of genomic and proteomic technology platforms used to generate drug response data in critical model systems; (B) animal handling, treatment and other experimental design optimizations; (C) data analysis strategies for extracting and validating candidate pharmacogenomic markers; and (D) overarching limitations in applying expression pharmacogenomics to animal model systems. PMID- 11913727 TI - The molecular dissection of human diseases after the human genome project. PMID- 11913728 TI - Polymorphisms in the ABC drug transporter gene MDR1. AB - In addition to genetically variable metabolic enzymes such as Cyp p450 proteins, blood and tissue levels of many drugs are influenced by controlled transport across compartmental boundaries. Major determinants in these transport processes are ATP-dependent efflux pumps such as P-glycoprotein and related proteins (eg MRPs), which can influence the bioavailability and CNS concentrations, as well as disposition of drugs. In addition to its recognized role in the development of multiple chemotherapy resistances, experimental evidence for the relevant influence of the MDR1 gene encoded P-glycoprotein, on the pharmacology of many other drugs has been gathered by the analyses of knockout mice, as well as in clinical studies. Recently, functional genetic polymorphisms in the MDR1 gene have been identified which influence the distribution and bioavailability of PGP substrates. PMID- 11913729 TI - Postgenomic prospects of success in drug development and pharmacotherapy. PMID- 11913730 TI - Thymidylate synthase gene polymorphism determines response and toxicity of 5-FU chemotherapy. AB - Thymidylate synthase (TS) catalyses the conversion of deoxy-uridylate to deoxy thymidylate and is essential for DNA synthesis. The human TS gene promoter is polymorphic, having either double or triple tandem repeats of a 28-bp sequence. Here we determined the significance of this polymorphism in humans and its prediction for clinical outcome of patients with metastatic colorectal cancer treated with 5-fluorouracil. The TS mRNA level was analyzed using RT-PCR. Individuals homozygous for the triple repeat variant (L/L) had 3.6 times higher TS mRNA levels compared to those homozygous for the double repeat variant (S/S) in tumor tissue (P = 0.004). We tested 50 patients with disseminated colorectal cancer who received 5-FU treatment to determine whether this TS polymorphism will predict clinical outcome. We found individuals with S/S genotype had a response rate of 50% (4/8) when compared to 9% (2/22) in those with L/L and 15% (3/20) in those with S/L genotype (P = 0.041). Patients with L/L had less severe side effects to 5-FU (P = 0.008). The data suggest that genotyping for the TS polymorphism may have the potential to identify patients more likely to respond to 5-FU based chemotherapy. PMID- 11913731 TI - Serotonin transporter gene associated with lithium prophylaxis in mood disorders. AB - The aim of this study was to investigate the possible association between the functional polymorphism in the upstream regulatory region of the serotonin transporter gene (5-HTTLPR) and the prophylactic efficacy of lithium in mood disorders. Two hundred and one subjects affected by bipolar (n = 167) and major depressive (n = 34) disorder were followed prospectively for an average of 58.2 months and were typed for their 5-HTTLPR variant using polymerase chain reaction techniques. 5-HTTLPR variants were associated with lithium outcome (F = 5.35; df = 2,198; P = 0.005). Subjects with the s/s variant showed a worse response compared to both l/s and l/l variants. Consideration of possible stratification effects such as sex, polarity, age at onset, duration of lithium treatment and previous episodes did not influence the observed association. 5-HTTLPR variants may be a possible influencing factor for the prophylactic efficacy of lithium in mood disorders. PMID- 11913732 TI - G649, an allelic variant of the human H2 receptor with low basal activity, is resistant to upregulation upon antagonist exposure. AB - Orange et al reported an allelic variant of the human histamine H2 receptor, in which adenine 649 was replaced with guanine, to be more frequent in the schizophrenic population than controls in British Caucasians. The A649 to G change causes an Asn to Asp transition at amino acid position 217 in the third intracellular region, which is postulated to be important for receptor function. Herein, we analyzed the functional significance of this variant using wild-type and variant receptors expressed in Chinese hamster ovary cells. The variant receptor was associated with markedly lower basal cAMP productions than the wild type receptor. Histamine-dependent cAMP productions via the variant receptor were lower as well. Treatment of cells expressing variant receptors with 10(-5) M ranitidine for 24 h resulted in a reduced degree of receptor upregulation as compared with the wild-type receptor. Thus, this is the first report of an allelic variant of the human H2 receptor which confers altered receptor function. To analyze gastric acid secretion in individuals with this variant, we examined 100 Japanese control subjects. However, neither heterozygotes nor homozygotes were found, suggesting that this variant, if present, is uncommon in the Japanese population. PMID- 11913733 TI - Sudden impact? The human genome sequence and the pace of gene discovery in complex diseases. PMID- 11913734 TI - Skin manifestations of internal malignancy. AB - This article concentrates on the major signs and syndromes that are associated with internal malignancies in the geriatric population. Included are cutaneous metastases, ectopic adrenocorticotropic hormone-producing syndromes, and disorders arising from APUD cell tumors. The major paraneoplastic disorders of dermatomyositis, generalized pruritus, Bazex's syndrome, and acanthosis nigricans also are discussed. Also included are Bowen's disease of skin; arsenical toxicity; and the Peutz-Jeghers', Gardner's, and Torre's syndromes, which are indicative of systemic or organ-related carcinogens. PMID- 11913735 TI - The aged epidermal permeability barrier: basis for functional abnormalities. AB - Aged epidermis develops an abnormality in permeability barrier homeostasis, which is accentuated further in photoaged skin. The biochemical basis is a global reduction in stratum corneum lipids and profound abnormality in cholesterol synthesis. Various cytokine/growth factor signaling pathways are abnormal in aged skin, particularly in the interleukin-1 family. Barrier repair therapy can be effective in restoring normal function if a cholesterol-dominant mixture of the three key physiologic lipids, including ceramides and free fatty acids, is emphasized. PMID- 11913737 TI - Drug eruptions and other adverse drug effects in aged skin. AB - The elderly are at higher risk for suffering the annoying and hazardous skin reactions that are associated with drug therapy. If a serious reaction occurs, the aged are also at higher risk for major morbidity and mortality compared with younger individuals. Early consideration of a drug cause and prompt cessation of all potentially associated drugs may improve a patient's outcome. Thus, a prompt, careful, and accurate characterization of a drug-related reaction is important in optimizing patient care, along with close monitoring for associated internal toxicities and other medical complications of severe cutaneous reactions. PMID- 11913738 TI - Bullous diseases in the elderly. AB - The elderly are prone to several autoimmune bullous diseases that have significant morbidity, and an accurate diagnosis is essential for proper management. It is important to be aware of the potential adverse effects of the various systemic agents that can be used. PMID- 11913736 TI - Hair loss and hirsutism in the elderly. AB - This article contains a brief review of hair follicle biology, followed by a presentation of the workup of elderly patients who present with hair loss or hirsutism. Common hair disorders, such as graying, telogen effluvium, androgenic alopecia, senescent alopecia, alopecia arcuata, hirsutism, and hypertrichosis, are discussed. PMID- 11913739 TI - Cutaneous fungal infections in the elderly. AB - Fungal infections are common in all age groups, including the elderly. In the elderly patient, unique challenges may exist in the diagnosis and treatment of these diseases. However, proper therapy for these prevalent conditions leads to a better quality of life. PMID- 11913740 TI - Leg ulcers. AB - Skin ulcers of the lower extremities have a chronic, relapsing course and often are difficult to manage. Leg ulcers remain a significant management challenge to health care professionals. This article focuses on the various causes of leg ulcers, with an emphasis on venous disease. Characteristic clinical findings and appropriate treatment options are discussed. Finally, the latest advances in the treatment of venous ulcers and their implications for clinical practice are emphasized. PMID- 11913741 TI - Skin infections and infestations in geriatric patients. AB - Geriatric patients develop infections, but many have a different appearance from what usually is expected. The difference depends on the age and immune status of the patient and the virulence of the organism. Differences may make recognition more difficult. Therapy may require different doses. Examples of the more common infections are detailed in this article. PMID- 11913742 TI - Working memory and situation model updating. AB - Situation model updating requires managing the availability of information as a function of its relevance to the current situation. This is thought to involve some aspect of working memory. The present study assesses the relation between updating ability and various measures of working memory span or capacity. In addition, a primitive general measure of situation model processing, a situation model identification test, and its relation to updating ability was also assessed. The present experiment used a version of a paradigm developed by Glenberg, Meyer, and Lindem (1987) to assess updating. Although updating was observed in both anaphoric reading time and recognition test accuracy measures, the reading time measure was relatively weak. Importantly, the updating effect on the recognition test was unrelated to working memory capacity. In contrast, updating was related to performance on the situation model identification task. Specifically, people who were good at model processing were better able to keep associated objects available than were people who were less adept. There were no differences in the maintenance of dissociated objects. These results suggest that the relationship between situation model processing and working memory capacity is relatively weak. PMID- 11913743 TI - The effects of readers' goals on inference generation and memory for texts. AB - We investigated the effects of readers' goals on inference generation and memory for expository text. College students (N = 82) read texts for the purpose of either study or entertainment. On-line inference generation was recorded via think-aloud procedures, and off-line memory was assessed via free recall. Reading goal strongly influenced inferential activity: Readers with a study goal produced more coherence-building (i.e., backward/explanatory and forward/predictive) inferences, whereas readers with an entertainment goal produced more associations and evaluations. These differences were associated with superior memory for the texts in the study condition. The results indicate that inference generation during reading is partly strategic and is influenced systematically by reading purpose. We propose that reading goals influence readers' standards of coherence, which in turn influence the types of inferences that they draw and the final memory representations that they construct. PMID- 11913744 TI - Letter detection in very familiar texts. AB - In the present study, we investigated whether patterns of letter detection for function and content words in texts are affected by the familiarity of the material being read. In Experiment 1, subjects searched for target letters in sentences that had been rehearsed prior to performing the letter detection on them as well as on unfamiliar sentences. In Experiment 2, subjects searched for target letters in highly familiar verses (e.g., nursery rhymes) and in unfamiliar sentences that were matched to the familiar verses. A disadvantage in letter detection for function as compared with content words consistently found with unfamiliar passages was reduced significantly with the familiar material in both experiments. Specifically, letter detection for content words grew worse in familiar text, but letter detection for function words showed a contrasting modest, though nonsignificant, improvement. The results are consistent with the proposition that in very familiar texts, parafoveal analysis permits the identification of generally less familiar content words. Simultaneously, the normal pattern of weighing the structure and content elements of text changes so that more fixations on function words occur than when one is reading unfamiliar texts. PMID- 11913745 TI - When being included is being excluded: a note on complement set focus and the inclusion relation. AB - Some negative quantifiers lead to pronominal reference patterns that are different from those obtained with positive quantifiers (Moxey & Sanford, 1993). This has been interpreted as meaning that the negatives give rise to a focus on the complement set (Moxey & Sanford, 1987); so, given few of the children enjoyed the trip, focus is on those who did not enjoy the trip. To date, this interpretation has depended on subjective judgments as to which set an anaphoric plural pronoun is referring to, allowing other interpretations of the data to be given by discourse semanticists. In two studies, we use the attachment patterns associated with the expression including, thereby circumventing the judgment problem. We show that a case like not many people enjoyed the race, including John leads to a representation in which John maps into the set of individuals who did not enjoy the race. We test and support the earlier claim that complement set focus is driven by denials associated with some negative quantifiers. PMID- 11913746 TI - Divided attention and prerecognition processing of spoken words and nonwords. AB - Three recognition memory experiments examined phonemic similarity and false recognition under conditions of divided attention. The manipulation was presumed to have little effect on automatic, perceptual influences of memory. Prior research demonstrated that false recognition of a test word (e.g., discrepancy) was higher if the study list included a nonword derived from the future test word by changing a phoneme near the end of the item (e.g., discrepan/l/y) relative to an early phoneme change (e.g., /l/iscrepancy). The difference has been attributed to automatic, implicit activation of test words during prerecognition processing of related nonwords. Three experiments demonstrated that the late-change condition also contributed to higher false recognition rates with divided attention at encoding. Dividing attention disrupted recognition memory of studied words in Experiments 1 and 3. Results are discussed in terms of their relevance for an interpretation emphasizing the automatic, implicit activation of candidate words that occurs in the course of identifying spoken words and nonwords. PMID- 11913747 TI - Nonconscious temporal cognition: learning rhythms implicitly. AB - Two experiments demonstrate that people can implicitly learn rhythms. Participants responded to a series of fast-paced beeps by pressing a key as soon as possible after each beep. They were not told that the duration (180, 450, or 1,125 msec) between each keypress and the next beep was specified by a repeating sequence. In both experiments, participants responded significantly faster to predictable, sequenced timing than to random timing but did not show more knowledge of the sequence than did control participants on explicit memory measures. This dissociation was obtained even with an explicit memory test in Experiment 2 that maintained the same context and response metric as the implicit task to maximize the transfer of relevant knowledge. Implications for temporal cognition are discussed. PMID- 11913748 TI - Intended and unintended effects of explicit warnings on eyewitness suggestibility: evidence from source identification tests. AB - Previous studies have shown that source identification (ID) tests reduce, and in some cases eliminate, eyewitness suggestibility errors. The present study showed that the suggestibility errors participants committed on a source ID test were further reduced when they were given the explicit postwarning that the experimenter was trying to trick them. These postwarnings reduced suggestibility to the same extent as prewarnings, and they did so for both once and repeatedly suggested items. In addition, the benefits of the pre- and postwarnings persisted when participants were retested 1 week later, but only if the suggestions had been repeated. For once-suggested items, the warning had the unintended effect of improving old/new recognition of the suggested information at retest, an effect that offset the improvements in source discrimination accuracy conferred by the warning. The advantages of using source ID tests for investigating group differences in eyewitness suggestibility are discussed. PMID- 11913749 TI - Interrupting recognition memory: tests of familiarity-based accounts of the revelation effect. AB - The revelation effect is a puzzling phenomenon in which items on a recognition test are more likely to be judged as "old" when they are immediately preceded by a problem-solving task, such as anagram solution. The present experiments were designed to evaluate Westerman and Greene's (1998) and Hicks and Marsh's (1998) familiarity-based accounts of this effect. We found comparable revelation effects when probes were preceded by an anagram or a numerical addition task and when subjects performed either one or two of these tasks. Taken together, the results do not support familiarity-based accounts of the revelation effect but are consistent with a proposed decision-based interpretation (i.e., criterion flux), in which it is assumed that the revelation task displaces the study list context in working memory, leading subjects to adopt a more liberal recognition decision criterion, thereby increasing the hit and false alarm rates. PMID- 11913750 TI - Forming and canceling everyday intentions: implications for prospective remembering. AB - The intention superiority effect (ISE) is characterized by faster response time to task material intended for future performance than to neutral material with no associated intention or material that is linked to a canceled intention. The existence of the ISE has been explored here under naturalistic conditions in which participants self-initiate an intention that is of personal relevance to them. Participants were required to remember prospective tasks that were presented under the guise of preparatory tasks for the next participant. After encoding a pair of tasks, they were informed that one task no longer needed to be performed. Subsequent lexical decision data exhibited the expected effect of faster response time for intended items than for canceled items (experimental groups in Experiments 1A and 1B). No differences in response time were observed between two sets of canceled items (control group in Experiment 1A). When an intention coexisted with the expectation that a written description of the task would be available, no reliable difference in latencies for these items and canceled items was observed (control group in Experiment 1B). The results are discussed in terms of facilitatory and inhibitory processes that may allow us to contend with many intentions in everyday scenarios. PMID- 11913751 TI - The perceptual aspect of skilled performance in chess: evidence from eye movements. AB - Expert and intermediate chess players attempted to choose the best move in five chess positions while their eye movements were monitored. Experts were faster and more accurate than intermediates in choosing the best move. Experts made fewer fixations per trial and greater amplitude saccades than did intermediates, but there was no difference in fixation duration across skill groups. Examining the spatial distribution of the first five fixations for each position by skill group revealed that experts produced more fixations on empty squares than did intermediates. When fixating pieces, experts produced a greater proportion of fixations on relevant pieces than did intermediates. It is argued that expert chess players perceptually encode chess configurations, rather than individual pieces, and, consequently, parafoveal or peripheral processing guides their eye movements, producing a pattern of saccadic selectivity by piece saliency. PMID- 11913752 TI - Expanding the search for a linear separability constraint on category learning. AB - Formal models of categorization make different predictions about the theoretical importance of linear separability. Prior research, most of which has failed to find support for a linear separability constraint on category learning, has been conducted using tasks that involve learning two categories with a small number of members. The present experiment used four categories with three or nine patterns per category that were either linearly separable or not linearly separable. With overall category structure equivalent across category types, the linearly separable categories were found to be easier to learn than the not linearly separable categories. An analysis of individual participants' data showed that there were more participants operating under a linear separability constraint when learning large categories than when learning small ones. Formal modeling showed that an exemplar model could not account for many of these data. These results are taken to support the existence of multiple processes in categorization. PMID- 11913754 TI - Interrupting recognition memory: tests of a criterion-change account of the revelation effect. AB - The revelation effect is evidenced by an increase in positive recognition responses when the test probe is immediately preceded by an unrelated problem solving task. As an alternative to familiarity-based explanations of this effect (Hicks & Marsh, 1998; Westerman & Greene, 1998), Niewiadomski and Hockley (2001) proposed a decision-based account in which it is assumed that the problem-solving task displaces the study list context in working memory, leading subjects to adopt a more liberal recognition criterion. In the present study, we show that the revelation effect is seen when the stimulus materials are pure lists of very rare words or nonwords. In contrast, for mixed lists of common words and very rare words or nonwords, the revelation effect is found for common words but disappears for very rare words and nonwords. We argue that, in mixed lists, the liberal decision bias following the revelation task and the criterion changes between common words and very rare words and nonwords serve to offset each other. PMID- 11913753 TI - Category variability, exemplar similarity, and perceptual classification. AB - Experiments were conducted in which observers learned to classify simple perceptual stimuli into low-variability and high-variability categories. Similarities between objects were measured in independent psychological-scaling tasks. The results showed that observers classified transfer stimuli into the high-variability categories with greater probability than was predicted by a baseline version of an exemplar-similarity model. Qualitative evidence for the role of category variability on perceptual classification, which could not be explained in terms of the baseline exemplar-similarity model, was obtained as well. Possible accounts of the effects of category variability are considered in the General Discussion section. PMID- 11913755 TI - Searching for two things at once: evidence of exclusivity in semantic and autobiographical memory retrieval. AB - We examined whether retrieval from semantic memory (Experiment 1) and autobiographical memory (Experiment 2) is exclusive, or whether people can search for two things at once. In Experiment 1, participants retrieved items as quickly as possible over 4 ruin from single categories (e.g., foods, countries) and from disjunctive categories (e.g., foods or countries). In Experiment 2, participants retrieved autobiographical episodes associated with single cue words (e.g., flower, ticket) or with disjunctive cue words (e.g., flower or ticket). In both experiments, retrieval of items from the disjunctive category did not exceed predictions based on optimal sequencing of retrieval from the corresponding two single categories. That is, exclusivity was observed to occur in retrieval from among multiple nonoverlapping categories in both semantic and autobiographical memory. PMID- 11913756 TI - Reasoning counterfactually: combining and rending. AB - Counterfactual reasoning occurs when people are asked to assume for the sake of argument that a fact they previously thought was true is now false and to draw a conclusion on that basis. To accomplish this sort of reasoning requires a revising of one's beliefs, which was simulated in the present study. Students were shown a set of statements that they were to assure themselves was consistent. They were then asked to accept a counterfactual assumption as true and reconcile resulting inconsistencies among the set of statements. In these problems, one statement is a generality (e.g., All trees on the plaza are elms), another is a particular (e.g., This tree is a pine), and one is a counterfactual (e.g., Assume this tree is on the plaza). Students preferred to reconcile the inconsistency by identifying the generality as "true" and the particular as "false." They did this more often when the assumption combined categories than when it dislodged categories and when real beliefs were at stake rather than arbitrary generalities. This study tested current models of inference for their ability to account for counterfactual reasoning and found the results to be consistent with natural deduction system, mental models, and conceptual integration network approaches to everyday reasoning. PMID- 11913757 TI - Topic-vehicle interaction in metaphor comprehension. AB - Property comparison models of metaphor comprehension assume that the topic and vehicle terms in metaphors are both understood to be referring to their conventional literal referents. In contrast, the interactive property attribution model (Glucksberg, McGlone, & Manfredi, 1997) assumes that the vehicle is understood to be referring to a metaphoric category that includes the topic's literal referent as a member. A priming paradigm was used to test the implications of these different models. Prior to interpreting a metaphor, participants read (1) the topic or vehicle concept alone, (2) a sentence ascribing a metaphor-relevant property to one concept, or (3) a sentence ascribing a metaphor-irrelevant property to one concept. All of the prime types facilitated metaphor comprehension with the exception of sentences ascribing metaphor-irrelevant properties to vehicles. The failure of these sentences (but not their topic counterparts) to facilitate metaphor comprehension is attributable to their priming an inappropriate literal interpretation of the vehicle term. These results are consistent with the claim that irrelevant information is suppressed during language comprehension (Gernsbacher, 1990) and support the interactive property attribution model. PMID- 11913758 TI - A comparison between Oriental & American science education. PMID- 11913760 TI - New nystatin polymeric complexes and their in vitro antifungal evaluation in a model study with Fusarium oxysporum. AB - Six water-soluble nystatin-polyvilnylpyrrolidone complexes with respective MW of 10 kDa (NC1), 25 kDa (NC2), 30 kDa (NC3), 40 kda (NC4), 90 kDa (NC5), 360 kDa (NC6) were synthesized. The activity of the complexes was compared with that of nystatin against growth and spore germination of Fusarium oxysporum f.sp. radiciscucumerinum. The ED50 value (effective dose) of free nystatin in aqueous solution on growth inhibition on solid medium was determined at 35.7 ppm. The ED50 of the complexes NC3, NC4, NC5, and NC6 ranged from 2.2 to 4 times lower than that of nystatin. The NC6 complex exhibited the highest activity, followed by NC5, NC4, and NC3. The activities of NC1 and NC2 were about 3 and 1.7 times higher than nystatin respectively in the same in vitro model. The complexes NC6. NC1 and NC4 were 25.4, 13.6 and 6.9 times more active respectively than nystatin against spore germination of E oxysporum. The activity of the nystatin complexes was dependent on the molecular weight of the polymeric carrier. PMID- 11913759 TI - Mycetoma caused by Fusarium solani with osteolytic lesions on the hand: case report. AB - Eumycetoma is a mycotic disease caused by saprophytic soil fungi that are usually inoculated through minor injuries. A case of mycetoma in a Brazilian farmer aged 71 years is reported. This patient presented erythema and edema on the dorsal surface of the left hand with multiple crusted and cicatricial lesions. No macroscopic grains were observed. The histopathological findings showed grains consisted of numerous hyphae which stained well with Gomori-Grocott method. This material obtained by cutaneous biopsy was submitted to culture on Sabouraud's medium and the colonies were identified as Fusarium solani. The radiological studies revealed bone osteolytic lesions and the ultrasound showed pseudocysts and fistulae at the site of this infection. The patient was treated with oral ketoconazole with a good clinical response. PMID- 11913761 TI - Fungi isolated from skins and pens of healthy animals in Nigeria. AB - The mycoflora of 220 skin scrapings, hair, nail samples and pens' materials of apparently healthy animals including cows, sheep, goats, rabbits, pigs and dogs were determined. Twenty eight species of fungi belonging to ten genera were recovered. Chrysosporium spp. were the most common and C. keratinophilum was recovered from all animals. Dermatophytes which are known causal agents of dermatophytosis were also isolated in different frequencies (Microsporum gypseum, M. canis, Trichophyton mentagrophytes, T rubrum). PMID- 11913762 TI - The non-steroidal anti-inflammatory drug niflumic acid inhibits Candida albicans growth. AB - The non-steroidal anti-inflammatory drug niflumic acid was found to inhibit growth of the yeast form of Candida albicans. Niflumic acid inhibited respiratory oxygen uptake and it is hypothesised that this was achieved by cytosolic acidification and block of glycolysis. Inhibitory concentrations are compatible with current practice of topical application. PMID- 11913763 TI - Tinea cruris in female prostitutes. AB - Tinea cruris is a dermatophytosis that mainly affects males. Infections in females are rare, and there are no known data on the frequency of tinea cruris in female prostitutes. We describe seven female prostitutes with tinea cruris with on age range of between 19-34 years (mean 25.3). Each prostitute had a mean of 50 sexual partners per month. Four Trichophyton mentagrophytes (57.1%), 2 T rubrum (28.6%), and 1 Epidermophyton floccosum (14.3%) isolates were cultered. Tinea cruris transmission is mainly indirect, but direct contact may serve to transmit the disease in some cases, and the spread of active lesions in prostitutes through multiple contacts among with their clients is possible. PMID- 11913764 TI - Bulbs mycoflora and their relation with three stored product mites. AB - The distribution of moulds on stored and field onion and garlic plants infested by bulb mites in Assiut area (Egypt) was studied using PDA medium at 28 degrees C. Among 40 host samples and the three mite species tested no significant difference was noted in the contamination by moulds. A total of 20 species appertaining to 11 genera were identified from the tested mites and their habitats. The predominant moulds on all samples were "storage moulds" from the genera Aspergillus (A. niger, A. versicolor) and Penicillium (P. chrysogenum, P. funiculosum, and "field moulds" among which Alternaria, Cladosporium, Fusarium (and its teleomorphs) and Setosphaeria were encountered most frequently. One fungus well known facultative pathogen was obtained: Beauveria bassiana. The tested mites transfer A. niger, N. haematococca, R. stolonifer and P chrysogenum outside their bodies while, A. flavus and A. ochraceus transfer through their digestive tracts along with the foods. Individuals of all mites could survived till the end of the experiment on all fungal species tested except A. niger, A. ochraceus and A. sydowii. Among 48 isolates screened for their ability to produce chitinase, about 83% of the isolates could produce this enzyme. Most of the positive isolates (17 isolates) had moderate producers PMID- 11913766 TI - Cytotoxicity of trichothecenes and fusarochromanone produced by Fusarium equiseti strains isolated from Norwegian cereals. AB - The cytotoxicity and secondary metabolites of 28 Norwegian strains of Fusarium equiseti have been characterized. Trichothecenes and fusarochromanone (FUCH) in rice culture extracts of the strains were analysed by gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC). The following metabolites were found in all isolates: FUCH, nivalenol (NIV), scirpentriol (SCIRP), 4-acetylnivalenol (4-ac-NIV, also called fusarenon-X), 15 acetyl-nivalenol (15-ac-NIV), and diacetoxyscirpenol (DAS). 4,15-diacetyl nivalenol (diacetyl-NIV) was found in 5 isolates. Porcine kidney epithelial cells (PK15. American Type Culture Collection) were exposed to rice culture extracts to study cytotoxicity. Descriptive statistics and factor analysis of the identified secondary metabolites show that their main metabolites were FUCH, NIV, SCIRP, DAS and 15-ac-NIV, consecutively. The individual trichothecenes were highly intercorrelated, whereas the production of acetylated NIV and DAS was slightly less. Stepwise multiple regression analysis of cytotoxicity and metabolite profiles of rice culture extracts ascribed the toxicity mainly to a combination of FUCH and 15-ac-NIV, though SCIRP or DAS are agents in the combined toxicity as well. PMID- 11913765 TI - Association of aflatoxin biosynthesis and sclerotial development in Aspergillus parasiticus. AB - Secondary metabolism in fungi is frequently associated with asexual and sexual development. Aspergillus parasiticus produces aflatoxins known to contaminate a variety of agricultural commodities. This strictly mitotic fungus. besides producing conidia asexually, produces sclerotia, structures resistant to harsh conditions and for propagation. Sclerotia are considered to be derived from the sexual structure, cleistothecia. and may represent a vestige of ascospore production. Introduction of the aflatoxin pathway-specific regulatory gene, aflR, and aflJ, which encoded a putative co-activator, into an O-methylsterigmatocystin (OMST)-accumulating strain, A. parasiticus SRRC 2043, resulted in elevated levels of accumulation of major aflatoxin precursors, including norsolorinic acid (NOR), averantin (AVN), versicolorin A (VERA) and OMST. The total amount of these aflatoxin precursors, NOR, VERA, AVN and OMST, produced by the aflR plus aflJ transformants was two to three-fold that produced by the aflR transformants. This increase indicated a synergistic effect of aflR and aflJ on the synthesis of aflatoxin precursors. Increased production of the aflatoxin precursors was associated with progressive decrease in sclerotial size, alteration in sclerotial shape and weakening in the sclerotial structure of the transformants. The results showed that sclerotial development and aflatoxin biosynthesis are closely related. We proposed that competition for a common substrate, such as acetate, by the aflatoxin biosynthetic pathway could adversely affect sclerotial development in A. parasiticus. PMID- 11913768 TI - Dermal fibroblast morphology is affected by stretching and not by C48/80. AB - Both stretching and C48/80 have been hypothesized to cause disruption of cell matrix adhesions and thereby affect the dynamics of fluid balance in tissues. We investigated the effect of sinusoidal stretching and/or C48/80 on the morphology of fibroblasts in skin excised from the backs of Wistar-Moller rats in order to assess how these stimuli affect cellular interactions in tissues. Tissue samples were either soaked in Krebs' buffer with and without C48/80, or sinusoidally stretched (20% strain) in buffer with and without C48/80. Control skin was fixed immediately after excision. All tissues were processed for transmission electron microscopy. Morphometric analyses demonstrated that sinusoidal stretching of the skin results in the retraction or disruption of fibroblast cytoplasmic extensions, rounding up of the cell bodies and subsequently in increased tissue water content. C48/80 had no apparent effect on fibroblast morphology and adherence in tissues. PMID- 11913767 TI - Cloning of Aspergillus fumigatus histidine kinase gene fragment and its expression during invasive infection. AB - It was demonstrated recently that three histidine kinases genes in Candida albicans contributed to virulence, indicating the importance of signaling pathways regulated by histidine kinases. In the present study, using a set of degenerate primers, RT-PCR was performed with cDNA of A. fumigatus as a template. PCR products were cloned and sequenced. After Blast analysis, it was found that one fragment (named as AFHK1), 305 bp, was highly homologous to the two-component histidine kinase tesA gene of Aspergillus nidulans. But AFHKI was not completely identical to the FOS-1 gene of A. fumigatus. The same A. fumigatus strain was used to inoculate the mice for a murine model of invasive pulmonary aspergillosis (IPA). After 5-days post-inoculation, the lungs of infected animals were removed and incubated for 2 h at 37 degrees C in digestion buffer containing collagenase and trypsin. The pulmonary cells were removed by passing the suspension through a sieve. The non-filterable hyphae were treated with deoxygenated sodium cholate. Total RNA of A. fumigatus isolated from the infected tissues or cultured in vitro was extracted. With AFHKI as a probe. a Northern blot was performed. A 3.0 kb (approximate) transcript of mRNA was detected corresponding to the putative histidine kinase gene. It was demonstrated that that gene was expressed at markedly higher levels in vivo than in vitro. The results suggest that this gene may contribute to the survival and virulence of A. fumigatus. PMID- 11913769 TI - Transglutaminase participates in the incorporation of latent TGFbeta into the extracellular matrix of aging articular chondrocytes. AB - TGFbeta1 is a multifunctional peptide growth factor that promotes processes associated with age-related degenerative diseases in articular cartilage. Large quantities of TGFbeta1 are stored in cartilage extracellular matrix (ECM) in a latent form (LTGFbeta1), and yet little is known about the factors that participate in the incorporation of LTGFbeta1 into the highly specialized cartilage ECM. We previously demonstrated high levels of the protein cross linking enzyme transglutaminase (TGase) in aging articular chondrocytes and showed that this enzyme participated in LTGFbeta1 activation. This work explores the hypothesis that extracellular TGase participates in LTGFbeta1 incorporation into ECM in aging chondrocytes. We studied the effects of TGase inhibitors on TGFbeta1 levels in ECM of old and young porcine articular chondrocytes. TGase inhibitors decreased the quantity of LTGFbeta1 in the ECM in old but not in young chondrocytes to 60-70% of control values (p<.05). Fibronectin, an extracellular TGase competitive substrate, also decreased LTGFbeta1 levels in ECM (p<.01). Levels of activated TGFbeta1 also decreased in the presence of TGase inhibitors, as did levels of latent TGFbeta binding protein 1 in the cell layer. Extracellular TGase activity was present in old but not young chondrocyte cultures. These findings support a role for extracellular TGase in the incorporation of LTGFbeta1 in the ECM of aging chondrocytes. PMID- 11913770 TI - Ovotransferrin is a matrix protein of the hen eggshell membranes and basal calcified layer. AB - The eggshell is an highly ordered structure deposited in the distal oviduct and composed of calcium carbonate and an organic matrix which is believed to influence its fabric. We have identified ovotransferrin as an 80 kDa matrix protein observed at high concentration in the uterine fluid at the initial stage of shell mineralization, by N-terminal sequencing and western blotting using monoclonal and polyclonal antibodies. It is present in extracts from demineralized eggshell and was localized by immunofluorescence in the eggshell membranes and mammillae, which are the sites of calcite nucleation. Northern blotting and RT-PCR demonstrated that ovotransferrin message was expressed in the proximal oviduct (magnum and white isthmus), and at a lower magnitude in the distal oviduct (red isthmus and uterus). Ovotransferrin was revealed by immunofluorescence in the tubular gland cells of the uterus. Calcium carbonate crystals grown in vitro in the presence of purified ovotransferrin showed large modifications of the calcite morphology. These observations and its presence in eggshell and membranes suggest a dual role for ovotransferrin, as a protein influencing nucleation and growth of calcite crystals and as a bacteriostatic filter to reinforce its inhibition of Salmonella growth in egg albumen. PMID- 11913771 TI - Effects of pulsed electromagnetic fields on human articular chondrocyte proliferation. AB - Low-energy, low-frequency pulsed electromagnetic fields (PEMFs) can induce cell proliferation in several cell culture models. In this work we analysed the proliferative response of human articular chondrocytes, cultured in medium containing 10% FBS, following prolonged exposure to PEMFs (75 Hz, 2.3 mT), currently used in the treatment of some orthopaedic pathologies. In particular, we investigated the dependence of the proliferative effects on the cell density, the availability of growth factors and the exposure lengths. We observed that PEMFs can induce cell proliferation of low density chondrocyte cultures for a long time (6 days), when fresh serum is added again in the culture medium. In the same conditions, in high density cultures, the PEMF-induced increase in cell proliferation was observed only in the first three days of exposure. The data presented in this study show that the availability of growth factors and the environmental constrictions strongly condition the cellular proliferative response to PEMFs. PMID- 11913772 TI - Collagen-binding domain of a Clostridium histolyticum collagenase exhibits a broad substrate spectrum both in vitro and in vivo. AB - The substrate spectrum of the tandem collagen-binding domain (CBD) of Clostridium histolyticumclass I collagenase (ColG) was examined both in vitro and in vivo. CBD bound to insoluble type I, II, III and IV collagens in vitro, and to skin, aorta, tendon, kidney, trachea and corneal tissues containing various types of collagen fibrils or sheets. CBD bound to all kinds of collagen fibrils regardless of their diameters and also bound to sheet-forming collagen in the glomerular basal lamina or Descemet's membrane of the cornea. This wide substrate spectrum expands possible applications of the drug delivery system we proposed previously (PNAS 95:7018-7023, 1998). Therapeutic agents fused with CBD will bind not only to subcutaneous tissues, but also to other tissues containing non-type I collagen. PMID- 11913773 TI - Immunohistochemical localization of components of the insulin-like growth factor system in human deciduous teeth. AB - To investigate the occurrence of components of the insulin-like growth factor (IGF) system during the resorption process of shedding human deciduous teeth, we investigated sections of 13 decalcified and paraffin-embedded deciduous teeth immunohistochemically with antibodies against IGF-I and -II, six IGF binding proteins (IGFBPs 1-6) and the IGF receptors IGF1R and IGF2R. The teeth were in different stages of resorption and all showed reparative cementum formation. It was found that acellular extrinsic fiber cementum, reversal lines and reparative cellular intrinsic fiber cementum were immunoreactive for both IGFs and various IGFBPs. Therefore, in human deciduous teeth, all subgroups of cementum, but not dentine, may represent sources of components of the IGF system. Odontoclasts did not carry IGFs or the IGF1R, but IGFBPs and the IGF2R. Therefore, these cells, in contrast to osteoclasts, may not respond to IGFs, but may be involved in the release and sequestration of IGFs from cementum during the resorption process. In contrast to odontoclasts, cementoblasts and periodontal ligament (PDL) fibroblasts carried IGF1R. The influence of the IGF system on the function of these cells with respect to periodontal matrix turnover and cementogenesis is discussed. On the behalf of the IGFBP immunoreactivities found, the PDL extracellular matrix can be considered to be a reservoir for IGF system components, where binding proteins may regulate IGF distribution and activity. PMID- 11913775 TI - Identification and characterization of two genes encoding the eukaryotic initiation factor 4A in rice. AB - A cDNA encoding eukaryotic translation initiation factor 4A (eIF4A) was isolated from a cDNA library of rice (Oryza sativa L.). Based on this DNA sequence, a 414 amino acid protein exhibiting 67, 64 and 59% homology to the mouse, Schizosaccharomyces pombe and Saccharomyces cerevisiae eIF4A, respectively, was predicted. The deduced amino acid sequence contains the characteristic motifs shared by the DEAD box supergene family. Another cDNA of rice eIF4A was reported previously. Comparison of the coding sequences of the two rice eIF4As showed 85% homology in the nucleotide sequence and 90% homology in the amino acid sequence. The genomic clones corresponding to the two rice eIF4A cDNAs were also isolated from a genomic library of rice (Oryza sativa L.). It was found that the two genes have common patterns of exon-intron boundaries. Their coding regions are split into four exons, and there is an additional exon in the 5'-non coding region. PMID- 11913774 TI - Alterations of collagen matrix in weight-bearing bones during skeletal unloading. AB - Skeletal unloading induces loss of bone mineral density in weight-bearing bones. The objectives of this study were to characterize the post-translational modifications of collagen of weight-bearing bones subjected to hindlimb unloading for 8 weeks. In unloaded bones, tibiae and femurs, while the overall amino acid composition was essentially identical in the unloaded and control tibiae and femurs, the collagen cross-link profile showed significant differences. Two major reducible cross-links (analyzed as dihydroxylysinonorleucine and hydroxylysinonorleucine) were increased in the unloaded bones. In addition, the ratios of the former to the latter as well as pyridinoline to deoxypyridinoline were significantly decreased in the unloaded bones indicating a difference in the extent of lysine hydroxylation at the cross-linking sites between these two groups. These results indicate that upon skeletal unloading the relative pool of newly synthesized collagen is increased and it is post-translationally altered. The alteration could be associated with impaired osteoblastic differentiation induced by skeletal unloading that results in a mineralization defect. PMID- 11913776 TI - Genomic structure of mouse copper chaperone, COX17. AB - Coxl7p was first cloned as a cytoplasmic copper chaperone from yeast mutant and recent works suggested the existence of mammalian homologues. Previous report has shown that a gel filtration fraction of heart extract containing porcine Coxl7p peptide promoted the survival of NIH3T3 fibroblast cells. In the present study, we first cloned DNA fragments of the mouse COX17 gene. The mouse COX17 spans approximately 6kb and consists of three exons. It was mapped to the center of chromosome 16, using a radiation hybrid-mapping panel. The major transcription start site is 80 bp upstream of the ATG initiation codon as determined by rapid amplification of cDNA ends (5'-RACE) analysis. Two potential polyadenylation sites are 3233 and 3293 bp downstream of the termination codon, respectively. Transient transfection of reporter plasmids containing portions of the mouse COX17 5'-flanking region into AtT-20 and NIH3T3 cells allowed the localization of the essential promoter to a 0.8 kb region upstream of the transcription starting site. Furthermore, the transfected luciferase activity was much higher in AtT-20 than NIH3T3. According to sequence analysis of the approximately 0.8kb 5' flanking region, GC rich segments including consensus sequences for binding of the transcription factor Sp1, but no TATA/CAAT boxes, exist in the region of the transcription start site. Besides the GC box, binding sites for NRF-1 and 2 known as specific transcription factors for COX subunits are also localized around the transcription starting site. PMID- 11913777 TI - Cloning of bovine eukaryotic translation initiation factor 4E (eIF-4E) and its expression in the bovine mammary gland at different physiological stages. AB - Bovine eIF-4E cDNA was cloned and its expression in the bovine mammary gland at distinct physiological stages was investigated. Bovine eIF-4E cDNA and protein sequences are highly homologous to those from other mammals. Using Northern blot analysis, we did not detect eIF-4E expression in the prepubertal bovine mammary gland, whereas a low level of eIF-4E mRNA was observed in the mammary tissues of heifers during the third estrous cycle. The eIF-4E mRNA level was significantly higher in the lactating mammary gland compared to the mammary tissues obtained from heifers during the third estrous cycle. Alteration of eIF-4E expression in bovine mammary tissues during different physiological stages indicates the involvement of eIF-4E in mammary development. Elevated eIF-4E expression during lactation may be related to increased translation of certain mRNA or the acceleration of overall protein synthesis. PMID- 11913778 TI - Nucleotide sequence analysis of a transforming gene isolated from nasopharyngeal carcinoma cell line CNE2: an aberrant human immunoglobulin kappa light chain which lacks variable region. AB - A transforming gene, designated Tx, was isolated from a human nasopharyngeal carcinoma (NPC) cell line CNE2 by transfection and molecular cloning techniques. The Tx gene was analyzed using computer-based bioinformatics and compared with the known sequences in EMBL and GenBank databases. We found that Tx contains human immunoglobulin kappa light chain constant region, five intact joining regions J1-J5, five recombination signal sequences and an N-segment besides classic regulatory sequences such as TATA boxes, CAAT boxes, poly A signals, etc. Interestingly, Tx also contains several binding sites for nuclear transcription factors such as NF-kappaB, NF-IL6, TFIID, etc. In conclusion, there are only several base pairs mutations or deletions compared with normal Ig K JC gDNA fragment. In all, Tx is an aberrant human immunoglobulin kappa light chain that contains the constant region, five joining regions, which lacks the variable regions. PMID- 11913779 TI - The gas vesicle gene (gvp) cluster of the cyanobacterium Pseudanabaena sp. strain PCC 6901. AB - A gene cluster located downstream from gvpA in the cyanobacterium Pseudanabaena sp. strain PCC 6901 has been cloned and sequenced. The three genes, orf1, gvpN and gvpJ, are consecutive with no intergenic region. In contrast to GvpN and GvpJ, which share high similarity at the amino acid level with their counterparts in other cyanobacteria and halophilic archaea, Orf1 is only 29% identical to the C-terminal part of GvpC from Anabaena flos-aquae and its sequence organization is reminiscent of the halophilic archaeal GvpC. PMID- 11913780 TI - Genomic organization of the murine G protein beta subunit genes and related processed pseudogenes. AB - The functional significance of heterotrimeric guanine nucleotide binding protein (G protein) for the many physiological processes including the molecular mechanisms of drug addiction have been described. In investigating the changes of mRNA expression after acute psychostimulant administration, we previously identified a cDNA encoding a G protein beta1 subunit (Gbeta1) that was increased up to four-fold in certain brain regions after administration of psychostimulants. The mouse Gbeta1 gene (the mouse genetic symbol, GNB1) was mapped to chromosome 4, but little was known of its genetic features. To characterize the GNB1 gene further, we have cloned and analyzed the genomic structures of the mouse GNBI gene and its homologous sequences. The GNBI gene spans at least 50 kb, and consists of 12 exons and 11 introns. The exon/intron boundaries were determined and found to follow the GT/AG rule. Exons 3-11 encode the Gbeta1 protein, and the exon 2 is an alternative, resulting in putative two splicing variants. Although intron 11 is additional for GNBI compared with GNB2 and GNB3, the intron positions within the protein coding region of GNB1, GNB2 and GNB3 are identical, suggesting that GNB1 should have diverged from the ancestral gene family earlier than the genes for GNB2 and GNB3. We also found the 5' truncated processed pseudogenes with 71-89% similarities to GNBI mRNA sequence, suggesting that the truncated cDNA copies, which have been reverse-transcribed from a processed mRNA for GNB1, might have been integrated into several new locations in the mouse genome. PMID- 11913781 TI - M13 cloning of mung bean nuclease digested PCR fragments as a means of gap closure within A/T-rich, genome sequencing projects. AB - Obtaining the complete DNA sequence of a genome is often not straightforward. After standard shotgun sequencing strategies have been employed there are often gaps remaining and these can be the most intractable regions, frequently containing repeat sequences, "uncloneable" sequences and/or regions of potential secondary structure or differential base composition. In genomes with a high A/T content, such as Plasmodium falciparum and Dictyostelium discoideum, solving these gaps is a particularly difficult problem as the sequences concerned are "fragile" and easily denatured, commonly uncloneable and have a paucity of good oligonucleotide priming sites. Reported here is a simple, yet reliable method for determining the sequence of A/T-rich gap-spanning PCR products. This method relies on the slippage of the specificity of mung bean nuclease so that it digests A/T-rich double-stranded DNA into a set of deletion fragments that can then be cloned into M13, sequenced and the original sequence assembled therefrom. PMID- 11913782 TI - Tissue-specific expression and splicing of the rat polycystic kidney disease 1 gene. AB - Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic potentially lethal human disorder and the polycystic kidney disease 1 (Pkd1) gene is accounted for 85-90% of these cases. We have obtained rat Pkdl cDNA sequence and characterized splicing of Pkdl RNA transcripts in normal rat tissues. Our sequence data revealed a high conservation of the Pkdl gene between rat and other species and mapped rat Pkdl to chromosome 10 in "tail-to-tail" orientation to the tuberous sclerosis 2 (Tsc2) gene. Pkdl was found ubiquitously expressed in the normal rat tissues and the brain had a complex pattern of exon 12 splicing. A novel splicing variant lacking entire exon 31, which occurs in rat and mouse but not in humans, was also identified. As the rat appears to be a valuable model for investigating polycystic kidney disease, the characterization of the rat Pkdl gene will help facilitate future studies to elucidate the molecular mechanisms of cystogenesis in this animal model. PMID- 11913783 TI - Cloning and expressionanalysis of two cotton (Gossypium hirsutum L.) genes encodingcell wall proline-rich proteins. AB - Two cotton (Gossypium hirsutum L.) genes, ghprp1 and ghprp2, encoding cell wall proline-rich proteins (PRPs) have been cloned and characterized. The ghprpl gene has an open reading frame (ORF) that encodes a PRP of 299 amino acids (aa), whereas the ghprp2 gene contains an ORF that codes for a 310-aa PRP. The GhPRP1 has an 80% identity in aa sequence with that of GhPRP2. Like other plant cell wall PRPs, both cotton PRPs have a hydrophobic signal peptide at their N-termini, followed by repeating peptide units. Northern blot analyses showed that the ghprpl gene is predominantly expressed in the fiber during the elongation stage of fiber development. Reverse transcription (RT)-PCR analysis showed that ghprpl is expressed in both fiber and root tissues, whereas ghprp2 is in roots only. The ghprpl gene was shown to be present in the A1, A2, D1 and D5 genomes of Gossypium by PCR amplification, whereas the ghprp2 gene is only present in the A1 and A2 genomes. The ghprpl gene was over-expressed in the yeast Pichia pastoris, and the expressed GhPRP1 protein was used as an antigen to raise polyclonal antibodies (anti-GhPRP1). Western analysis using the anti-GhPRP1 probe detected a major protein band (50 kDa) in 5-31-day postanthesis (DPA) fibers. However, the 50 kDa protein was absent in other cotton tissues. PMID- 11913784 TI - Genomic DNA sequences encoding Malus x domestica Borkh. "Akane", "Delicious" and Malus transitoria S-RNases. AB - All S-RNases in apple contained one intron at the same location within the hypervariable region. They code for 225-228 amino acids, however, the length of introns is variable and divided into three groups. The intron sequences of some S RNases within the same group showed an extremely high similarity as well as their coding sequences, suggesting that they were generated from the same origin. PMID- 11913785 TI - Genomic DNA sequence and transcription factor binding sites of mouse ninjurin. AB - The complete genomic DNA sequence of mouse ninjurin gene has been cloned and sequenced by screening a bacterial artificial chromosome (BAC) library of mouse 129/SvJ genomic DNA. The mouse ninjurin gene comprises four exons and the translatable sequences are included in the first three exons. The putative promoter region of the mouse ninjurin gene lacks the consensus "CAAT" or "TATA" sequence. Nonetheless, it has demonstrated the promoter activity in transient transfection experiment using the construct containing putative promoter sequence of mouse ninjurin and reporter gene. The nucleotide sequence of the putative promoter region shows 83% homology with the corresponding DNA sequence of human ninjurin gene that had been previously reported, and reveals a high degree of conservation between the two species. Analysis of the DNA sequence identified the putative promoters and the binding sites for a variety of transcription factors of mouse ninjurin. PMID- 11913786 TI - Isolation and characterisation of a cDNA for a novel small HSP from sunflower suspension cell cultures. AB - A full-length cDNA encoding a novel small heat shock protein (HaHSP17.9) was isolated from a cDNA library of sunflower (Helianthus annuus cv. Gloriasol). The deduced amino acid sequence exhibited high degree homology to the class I cytosolic sHSPs from other plant species, and contained all the conserved regions characteristic of this class of proteins. Northern analyses showed that the transcript homologous to HaHSP17.9 accumulates during heat shock in suspension cultured cells and in the different parts of sunflower seedlings. PMID- 11913787 TI - Cloning and sequence analysis of cDNA for heavy-chain ferritin from the Canis familiaris. AB - Ferritin serves as a storage protein for iron in animals. Complementary DNA encoding a heavy chain ferritin was cloned from the brain of Canis familiaris. The dog ferritin cDNA encodes a 182 amino acid that shows high levels of amino acid identity with vertebrate ferritins (90-98%). Near the cap region of the 5' untranslated region, the dog H-ferritin mRNA displays a 28-nucleotide sequence that is exactly conserved in the corresponding region of the human and pig H ferritin mRNA, thus making this sequence a prime candidate for involvement in the known translational regulation of H-ferritin by iron. PMID- 11913788 TI - Gene structure and alternative splicing of murine podocalyxin: a member of the CD34 sialomucin family. AB - Podocalyxin is a sialoglycoprotein of the glomerular podocytes, vascular endothelial cells, platelets, and hemopoietic stem cells. The function of podocalyxin is unknown, but it contains most of the protein bound sialic acid in the glomerulus and is considered vital in the structure and function of the glomerular filtration apparatus. The murine podocalyxin full-length cDNA has been determined and is 5,318 base pairs. The gene localizes to chromosome 6B1 by FISH analysis and contains eight major exons with one additional alternatively spliced exon. The alternatively spliced isoform of podocalyxin codes for a truncated intracellular domain and is expressed in a tissue specific pattern in parallel with full-length podocalyxin. The organization of the gene structure of murine podocalyxin is similar to the murine CD34 gene and suggests a distant evolutionary relationship to CD34. PMID- 11913789 TI - Molecular cloning and sequence analysis of the manganese catalase gene from Thermoleophilum album NM. AB - The manganese catalase gene (mnct) from Thermoleophilum album NM, a thermophilic bacterium, was cloned and its nucleotide sequence was analyzed. The gene consists of 885 bp (65.4% GC content) encoding 294 amino acids with a molecular mass of 32,500 Da. The deduced amino acid sequence shows similarities to those of Thermus species strain YS 8-13 (a thermophilic bacterium) and Bacillus halodurans (an alkaliphilic bacterium) with 61 and 54% identities, respectively. PMID- 11913790 TI - Molecular cloning of cDNA encoding polypeptide chain elongation factor 1alpha from goldfish (Carassius auratus). AB - The goldfish homologue of polypeptide chain elongation factor-1alpha (EF-1alpha) isolated from the ovary of the goldfish is described. The deduced amino acid sequence is highly homologous to EF-1alpha from other species. Analysis of its tissue distribution revealed a single 1.7 kilobase message ubiquitous among various tissues. PMID- 11913791 TI - Cloning of the proto-oncogene c-src from rat testis. AB - The cellular homolog of the oncogene v-src, the proto-oncogene c-src, was cloned from rat testis using a high stringency polymerase chain reaction. Rat c-src cDNA shared identity with chicken and mouse, and Rous sarcoma virus c-src and v-src, respectively. Rat c-Src protein was 98% homologous to both human and mouse c-Src. Interestingly, rat Src contained one extra amino acid compared to the mouse protein. As expected, the rat testis Src lacked the six extra residues common to the neuronal Src identified in human and mouse. Reporting of the cDNA sequence for non-neuronal, rat c-src should facilitate experimentation into cell growth and transformation using rat tissues as models of human disease. PMID- 11913792 TI - The gene causing the Best's macular dystrophy (BMD) encodes a putative ion exchanger. AB - Best's macular dystrophy (BMD), also known as vitelliform macular degeneration type 2, is an autosomal dominant disease that causes loss of vision. The causative gene encodes a 585 amino acids protein called bestrophin with unknown function. From bioinformatics analysis, a putative ion exchanger function for bestrophin can be suggested. PMID- 11913793 TI - Nucleotide sequence of envelope protein of Japanese encephalitis virus SA14-14-2 adapted to vero cells. AB - Live attenuated Japanese encephalitis (JE) virus SA(14)-14-2 produced in primary dog kidney cells (PDK) was adapted to Vero cells. In an effort to gain insight into the molecular basis of the biological characteristics of the SA14-14-2(Vero) strain, the 1500 nucleotide sequence encoding the envelope (E) gene which possesses major neutralizing epitopes was determined and compared with the sequences of two other attenuated JE virus strains, SA14-14-2(PHK) and SA14-14 2(PDK). The amino acid sequence of the C-terminal region (a.a. 280-500) was found to be identical for all three strains, while the N-terminal region (a.a. 1-279) shows sequence variation. The distribution of mutations in the N-terminal region was nearly the same among the three attenuated strains, suggesting that the N terminal sequences might be related with virus-host cell specificity. However, it was found that Lys and Val (a.a. 138 and 176, respectively), known to be responsible for attenuation, are still conserved in SA(14)-14-2(Vero). Animal testing showed that SA(14)-14-2(Vero) has an attenuation phenotype similar to that of the parent SA(14)-14-2(PDK) strain in mice. PMID- 11913794 TI - Nucleotide sequence of the porcine 60S ribosomal protein L35 cDNA. AB - The cDNA encoding the 60S ribosomal protein L35 was cloned from the porcine liver cDNA library using the N-terminal fragment coding region of human protein disulfide isomerase as the probe. PMID- 11913796 TI - Analysis and expression of algL, which encodes alginate lyase in Pseudomonas syringae Pv. syringae. AB - Pseudomonas syringae produces alginate, an exopolysaccharide that contributes to the virulence and epiphytic fitness of this phytopathogenic bacterium. P. syringae also produces the algL-encoded alginate lyase, which cleaves the alginate biopolymer via a beta-elimination reaction. The algL gene from P. syringae maps to a 1134 bp region within the alginate biosynthetic operon, and is similar to algL from Halomonas marina, P. aeruginosa, Azotobacter chroococcum, and A. vinelandii. algL from P. syringae was over expressed in Escherichia coli; two periplasmic forms of AlgL were overproduced (40 and 37 kDa). Both forms were enzymatically active and recognized by antibodies raised against AlgL from P. aeruginosa. Analysis of the regions flanking algL revealed significant homology to algX and algI, genes previously identified in the biosynthetic operon of other alginate-producing bacteria. PMID- 11913795 TI - Cloning and sequence determination of a gene encoding an osmotin-like protein from strawberry (Fragaria X ananassa Duch.). AB - Osmotin and osmotin-like proteins (OLPs) are pathogenesis-related (PR) proteins, whose synthesis is normally stimulated upon infection of plants by pathogens. A strawberry genomic clone containing an osmotin-like protein (OLP) gene was isolated and sequenced. This clone contains an open reading frame of 681 nucleotides without any intron. The predicted amino acid sequence of the protein shares high degrees of homology with a number of other OLPs and related proteins, of which several are known to have antifungal activities. Southern hybridization analysis of strawberry genomic DNA suggested that the OLP is coded by a multi gene family. Results from reverse transcriptase-polymerase chain reaction indicated that this OLP gene is expressed in uninfected strawberry plants. PMID- 11913797 TI - Molecular characterization of the Treponema denticola fliQ region. AB - A Treponema denticola 4.2 kb DNA region containing four complete genes (orfl, fliQ, fliR, and flhB) and a truncated gene (flhA') was sequenced and analyzed. The deduced amino acid sequences of FliQ, FliR, FlhB and FlhA' have significant homology with bacterial proteins associated with the flagellar export apparatus, whereas the deduced amino acid sequence of Orf1 has homology with an E. coli alcohol dehydrogenase. A putative sigma70-like promoter was identified upstream of fliQ. RT-PCR analysis indicated that fliQ, fliR, flhB and flhA' are co transcribed independently of orfl, suggesting that the motility-associated genes are components of an operon. The location of the T. denticola fliQ-flhA' genes differs from that of the corresponding T. pallidum and Borrelia burgdorferi genes which are present in the large fla or flgB flagellar operons, respectively. PMID- 11913798 TI - Interaction of phenol derivatives with ion channels. PMID- 11913799 TI - Sedation assessment in critically ill patients with bispectral index. AB - BACKGROUND AND OBJECTIVE: Clinical sedation assessment becomes insufficient in deeply sedated patients. Bispectral index as a processed electroencephalogram parameter provides a continuous and observer-independent value reported to change with sedation. The aim of this prospective observational study was to determine the reliability and possible confounding factors of the bispectral index to assess sedation in surgical intensive care patients. METHODS: Following major surgery, bispectral index, body temperature and electromyographic activity of 44 ventilated patients were recorded. Sedation levels were assessed with Ramsay sedation score. RESULTS: Although bispectral index correlated with Ramsay sedation score (-0.64; P < 0.01) we found that in deeply sedated patients temperature instability and electromyographic activity increased bispectral index. Bispectral index correlated significantly with electromyographic activity (0.80; P < 0.01) and with an increase of body temperature (0.55; P < 0.01) not only in all patients but also in clinically deeply sedated patients (0.57; P < 0.01 and 0.46; P < 0.05). CONCLUSIONS: Only under certain conditions, such as low muscular activity and body temperature stability, may the bispectral index be a useful addition to clinical scoring in the sedation assessment of critically ill patients. PMID- 11913801 TI - The effects of fenoldopam on renal function in patients undergoing elective aortic surgery. AB - BACKGROUND AND OBJECTIVE: Postoperative renal impairment is a recognized complication of infrarenal aortic cross-clamping. Our hypothesis was that the renal vasodilating and natriuretic effects of fenoldopam mesylate, a selective dopamine (DA1) agonist, would preserve renal function in patients undergoing elective infrarenal aortic cross-clamping. METHODS: A prospective, randomized, double blind controlled clinical trial was performed. Twenty-eight ASA II-III patients undergoing elective aortic surgery requiring infrarenal aortic cross clamping were studied. According to random allocation, patients received either fenoldopam (0.1 microg kg(-1) min(-1)) or placebo intravenously prior to surgical skin incision until release of the aortic clamp. Plasma creatinine, creatinine clearance, urinary output, fractional excretion of sodium, and free water clearance were measured: (a) prior to admission to hospital; (b) during the period from insertion of the urinary catheter until application of the aortic cross-clamp; (c) during the period of aortic cross-clamping; (d) 0-4 h, and (e) 4 8 h after release of the clamp and on days 1, 2, 3, and 5 postoperatively. RESULTS: Fenoldopam (0.1 microg kg(-1)min(-1)) administration was not associated with haemodynamic instability. On application of the aortic cross-clamp creatinine clearance decreased significantly in the placebo (83 +/- 20 to 42 +/- 29 mL min(-1) (mean +/- SD)) (P < 0.01) but not in the fenoldopam group, and this decrease persisted for at least 8 h after release of the cross-clamp (83 +/- 20 to 54 +/- 33 mL min(-1) (mean +/- SD)) (P < 0.05). Plasma creatinine concentration increased significantly from baseline on the first postoperative day in the placebo group (87 +/- 12 to 103 +/- 28 micromolL(-1) (mean +/- SD)) (P < 0.01) but not in the fenoldopam group. CONCLUSIONS: These findings are consistent with the hypothesis that fenoldopam possesses a renoprotective effect during and after infrarenal aortic cross-clamping. PMID- 11913800 TI - First clinical experience with the rapid-, short-acting amiodarone derivative E 047/1 after cardiac surgery. AB - BACKGROUND AND OBJECTIVE: Amiodarone is very effective against a variety of dysrhythmias but has poor pharmacodynamic properties and many undesired side effects. Its short- and rapid-acting derivative E 047/1 may circumvent some of these drawbacks. It is easier to titrate while retaining the high efficacy of amiodarone and may have acceptable influences on haemodynamics and cardiac conduction in patients who develop serious, destabilizing ventricular tachydysrhythmias after cardiac surgery. METHODS: Testing E 047/1 was performed prospectively in two consecutive phase II open, clinical studies. Out of 504 patients scheduled for surgery using cardiopulmonary bypass for coronary artery grafting and/or valve repair, 35 developed serious, haemodynamically destabilizing ventricular dysrhythmias (Lown 2-Lown 4b) after surgery and were treated with a 1 mg kg(-1) (pilot study, n = 15) or randomized to a 2 or 3 mg kg( 1) bolus of E 047/1, followed by a 1 mg kg(-1) h(-1) continuous infusion for 2 h (n = 10 in each group). Dysrhythmias, PQ, QTc intervals and haemodynamics using the thermodilution technique were evaluated for up to 24 h after drug initiation. RESULTS: At the time of final inclusion the patients had between 6 and 12 (or more) ventricular ectopics per minute. Within the first 2-3 min of application in the pilot trial E 047/1 induced a decrease of ventricular dysrhythmias to between 0 and 4 per min, a decrease that held for the duration of treatment. The area under the curve decreased from 434 (322, 855; median, quartiles) to 114 (9, 477, P < 0.01) events per hour. In the randomized trial, E 047/1 administered in either dose rapidly reduced ventricular dysrhythmias at least as effectively as in the pilot trial 565 (478, 701) to 33 (8, 238, P < 0.05) after a 2 mg bolus; 482 (339, 482) to 95 (13, 540, P < 0.01) events per hour after a 3 mg bolus. Approximately 4-6 h after drug termination, dysrhythmias reappeared in the majority of patients. In only three patients did the incidence of dysrhythmias return to inclusion criteria levels. In contrast to the pilot trial, in the randomized trial there was a slight increase of mean pulmonary artery pressure, central venous pressure and pulmonary arterial wedge pressure and a slight decrease of LCWI in both groups. E 047/1 did not cause QTc prolongation. CONCLUSIONS: E 047/1 appears to be a safe alternative to amiodarone in the perioperative setting of cardiac surgery when serious, destabilizing dysrhythmias occur. PMID- 11913803 TI - The prevention of propofol injection pain by tramadol or ondansetron. AB - BACKGROUND AND OBJECTIVE: To compare the efficacy of tramadol and ondansetron in minimizing the pain due to injection of propofol in 100 patients. METHODS: An intravenous cannula was inserted in the dorsum of the hand. After tourniquet application to the forearm, tramadol 50 mg (Group 1, n = 50) or ondansetron 4 mg (Group 2, n = 50) was injected. The tourniquet was released after 20 s, and propofol 5 mL was administered over 5 s. The patients were observed and asked if they had pain in the arm and the response was assessed. Nausea and vomiting and degree of sedation were recorded for the first postoperative 24 h. RESULTS: Twenty-one patients in Group 1 and 14 patients in Group 2 reported no pain. Slight pain was seen in 15 patients in Group 1 and in 18 patients in Group 2. Moderate pain was seen in 10 patients in Group 1 and 15 patients in Group 2. Severe pain was seen in four of the patients in Group 1 and three patients in Group 2. There was no significant difference of pain between Groups 1 and 2, but we found a significant reduction of nausea and vomiting in the ondansetron group compared with the tramadol group (P = 0.033). CONCLUSIONS: Tramadol or ondansetron are equally effective in preventing pain from propofol injection. The added benefit of a reduction in nausea and vomiting after operation in the ondansetron group may be a reason to prefer this drug. PMID- 11913802 TI - Haemodynamic changes in ischaemic vs. anhepatic pig experimental model of acute liver failure. AB - BACKGROUND AND OBJECTIVE: The removal of the non-functioning liver in cases of fulminant liver failure has been advocated by some authors as a means of improving haemodynamic instability and acid-base disturbances associated with acute liver failure. METHODS: The aim of the present experimental study was to investigate whether maintaining a non-functioning liver is preferable over removing it in terms of haemodynamic variables, after acute hepatic failure has been surgically induced. Twenty Landrace pigs were used in the study. All of them underwent portocaval anastomosis and ligation of the hepatic artery. After an 18 h period and with biochemical indices of fulminant hepatic failure clearly demonstrated, the animals were randomly assigned to one of two groups: in 10 pigs (Group A) the ischaemic liver was left in situ and no further surgical intervention was undertaken. The other 10 (Group B) underwent total hepatectomy. Haemodynamic monitoring was the same in both groups. No inotropes were administered throughout the whole period of observation. RESULTS: Haemodynamic deterioration was observed in the hepatectomized pigs (Group B) whereas the group with the ischaemic liver in situ (Group A) remained stable in terms of the haemodynamic variables evaluated until the end of the experiment. (Cardiac index in Group A 7.59 +/- 1.25 L min(-1) m(-2) vs. 2.92 +/- 0.68 L min(-1) m(-2) in Group B, P < 0.05.) CONCLUSIONS: The concept of salvage hepatectomy in cases of acute liver failure should be redefined since there seems to be some experimental evidence that it may not be as beneficial as originally thought. PMID- 11913804 TI - Magnesium infusion reduces perioperative pain. AB - BACKGROUND AND OBJECTIVE: Magnesium has antinociceptive effects in animal and human models of pain. These effects are primarily based on the regulation of calcium influx into the cell. The aim of this study was to determine whether perioperative infusion of magnesium would reduce postoperative pain and anxiety. METHODS: Twenty-four patients, undergoing elective hysterectomy, received a bolus of 30 mg kg(-1) magnesium sulphate or the same volume of isotonic sodium chloride solution intravenously before the start of surgery and 0.5 g h(-1) infusion for the next 20 h. Intraoperative and postoperative analgesia were achieved with fentanyl and morphine respectively. Patients were evaluated pre- and postoperatively for anxiety. RESULTS: Fentanyl consumption and total morphine requirements were significantly decreased in the magnesium group compared to the control group. Postoperative anxiety scores and sedation were similar between groups. CONCLUSIONS: Continuous magnesium infusion, including the pre-, intra-, and postoperative periods reduces analgesic requirements. These results demonstrate that magnesium can be an adjuvant for perioperative analgesic management. PMID- 11913805 TI - Perioperative risk factors in elective pneumonectomy: the impact of excess fluid balance. AB - BACKGROUND AND OBJECTIVE: This study was performed to identify risk factors for complications and in-hospital mortality associated with pneumonectomy. METHODS: The influence of fluid balance during anaesthesia was evaluated, taking into account the patient's age, gender and body mass index, smoking habits, history of pulmonary or cardiac disorders, the site of pneumonectomy and duration of anaesthesia. One-hundred-and-seven patients undergoing elective pneumonectomy were included in the study. RESULTS: A total of 31 patients (29%) suffered from one or more postoperative complications, seven (22.4%) of these had severe dysrhythmias, six (19.6%) had pulmonary complications and three (9.3%) had cardiovascular complications. The overall mortality rate was 10.3%. CONCLUSIONS: Based on logistic regression analysis, our data indicate the following risk factors for postoperative complications: positive fluid balance exceeding 4000 mL during anaesthesia (pulmonary complications and mortality), body mass index < 17 or > 25 kg m(-2) (severe dysrhythmias), or history of chronic heart disease (pulmonary complications). Thirteen patients (12.4%) suffered from a fluid balance > 4000 mL during anaesthesia. Regression analysis indicated that fluid balance exceeding 4000 mL was associated with a higher risk of postoperative complications than blood loss exceeding 1000 mL and to be the strongest risk factor for postoperative pulmonary complications and in-hospital mortality. Further trials estimating the effect of restrictive fluid regimens and the use of vasopressors for blood pressure control during anaesthesia must be carried out. PMID- 11913806 TI - Auditory function following spinal analgesia. Comparison of two spinal needles. AB - BACKGROUND AND OBJECTIVE: Auditory impairment is among the lesser known complications of spinal analgesia. The aim of the present study was to determine the degree of vestibulocochlear dysfunction in patients undergoing spinal analgesia for lower abdominal surgery. METHODS: Eighty patients who had received spinal analgesia for lower abdominal surgery were studied. Males were undergoing inguinal herniorraphy and the females tubectomy. Audiograms were performed before operation and on the second and seventh postoperative days. Hearing levels were measured from 250 Hz-8 kHz. In Group 1 (n = 40) a 22-gauge, cutting type of spinal needle (Howard Jones) was used. In Group 2 (n = 40) a 25-gauge, non cutting spinal needle (Whitacre) was used. RESULTS: Hearing loss >10 dB was noticed in three patients in Group 1 and none in Group 2. The mean hearing level was more reduced in Group 1 patients. CONCLUSIONS: Use of cutting type spinal needle is associated with a greater decrease in mean hearing levels compared to the non-cutting type. PMID- 11913807 TI - Another neurological complication of central venous cannulation. PMID- 11913808 TI - From which countries do chairpersons and invited speakers at important anaesthesia and intensive care meetings come? PMID- 11913809 TI - Anaesthetic agents in paediatric day case surgery: do they affect outcome? AB - Both the numbers of children undergoing day case surgery and the type of procedures performed in this way are increasing. This expansion will only be beneficial if anaesthesia and surgery are provided with minimal post-operative morbidity e.g. postoperative delirium or nausea and vomiting. The choice of anaesthetic technique is considered critical to optimizing the service provided to patients and for this reason much research has addressed this question. This review considers the effect of anaesthetic technique on postoperative outcome in paediatric day case surgery. The outcome measures reviewed by this article are induction of anaesthesia, effects on the cardiovascular system, recovery from anaesthesia and postoperative nausea and vomiting. In each section both quantitative and qualitative outcome measures are discussed. Comparisons are made between sevoflurane and halothane, sevoflurane and propofol, propofol and halothane, desflurane and halothane and the presence or absence of nitrous oxide. PMID- 11913810 TI - Honey bees store landmarks in an egocentric frame of reference. AB - Honey bees are well known to rely on stored landmark information to locate a previously visited site. While various mechanisms underlying insect navigation have been thoroughly explored, little is yet known about the degree of integration of spatial parameters to form higher-level spatial representations. In this paper we explore the basic interactions between landmark cues and directional cues, which stand at the basis of our understanding of piloting mechanisms. A novel experimental paradigm allowed us independent manipulation of each parameter in a highly controlled environment. The approach taken was twofold: cue-conflict experiments were first conducted to examine the interactions between positional cues and directional cues. The bees were then successively deprived of sensory cues to question the dependence of landmark navigation on context cues. Our results confirm previous findings that landmark cues are used in concert with external directional cues if present. Conversely, the bees' ability to locate a food site was not disrupted in the absence of an external directional reference. Thus, bees store landmark memories in an egocentric frame of reference and only loose and facultative associations between visual memories and compass cues are formed. PMID- 11913811 TI - Temporal modulation transfer functions in the barn owl (Tyto alba). AB - Barn owls (Tyto alba) have evolved several specializations in their auditory system to achieve the high sensory acuity required for prey capture, including superior processing of interaural time differences and phase coding in the auditory periphery. Here, we tested whether barn owls are capable of high temporal resolution that may be a prerequisite for the accuracy in binaural processing. Temporal resolution was measured psychoacoustically and demonstrated in temporal modulation transfer functions. Four barn owls were trained in an operant task with food reward to detect sinusoidal amplitude modulations within an 800-ms gated white-noise burst or 800-ms periods of modulation in continuous white noise (spectrum levels of -5 dB and 15 dB SPL). Within the range of tested amplitude modulation frequencies from 5 Hz to 1280 Hz, barn owls' detection thresholds were lowest at 10-20 Hz. This sensitivity corresponds to an intensity difference limen of between 0.9 dB and 1.4 dB. For all conditions, temporal modulation transfer functions showed band-pass characteristics with a high frequency cutoff in the range of 37 Hz to 92 Hz, corresponding to minimum integration times of 4.3 ms and 1.7 ms, respectively. In summary, these data indicate a temporal resolution in the owl's auditory system that is good, but not unusual, compared to other vertebrates. PMID- 11913812 TI - Effects of a hydroxy-cinnamoyl conjugate of spermidine on arthropod neuromuscular junctions. AB - N1-coumaroyl spermidine is structurally similar to acylpolyamines found in spider and wasp venoms, which are known to block arthropod glutamate receptors. N1 coumaroyl spermidine reduced the amplitude of excitatory postsynaptic potentials recorded in crayfish muscle. This effect was dose dependent, with an IC50 value of 70 micromol l(-1). N1-coumaroyl spermidine reversibly reduced the amplitude of potentials elicited by iontophoretic application of L-glutamate, indicating a direct effect on postsynaptic glutamate receptors. Neither 1 mmol l(-1) spermidine nor 1 mmol l(-1) coumaric acid altered excitatory postsynaptic potential amplitude, indicating that blockage requires the conjugated phenolic polyamine. N1-coumaroyl spermine, a slightly longer phenolic polyamine, reduced excitatory postsynaptic potential amplitude with approximately the same potency as N1-coumaroyl spermidine. Thus, potency of blockage does not appear to be affected in this experimental preparation by small changes in length of the polyamine. N1-coumaroyl spermidine also reduced excitatory postsynaptic potentials in muscles of the insect Drosophila. The ability of N1-coumaroyl spermidine to attenuate synaptic transmission at insect neuromuscular synapses lends support to the notion that plant-derived phenolic polyamines might serve as natural insecticides. PMID- 11913813 TI - Visual acuity thresholds of juvenile loggerhead sea turtles (Caretta caretta): an electrophysiological approach. AB - Visual evoked potentials measure dynamic properties of the visual system by recording transient electric responses of neural tissue identified to correspond to a specific visual stimulus, such as light or a striped grid. In this study, visual evoked potentials were used to test the visual acuity of juvenile loggerhead sea turtles (Caretta caretta) in water. Subject animals were fitted with a Plexiglas goggle filled with filtered seawater. Stimuli of black and white striped gratings were presented to the turtles using a slide projector directing an image onto a screen via a rotatable mirror that shifted the striped pattern laterally one-half cycle. Bioelectric activity was collected using a digital averaging computer and subdermal platinum electrodes, implanted under the head scutes directly above the optic nerve and the contralateral optic tectum. To isolate the response signal from the noise, signal averaging techniques were used when collecting visual evoked potentials. The resulting response waveforms included a robust positive-negative compound that was used to track the turtle's response to visual stimulation. Acuity thresholds for these sea turtles, which were derived from linear regressions analysis of the positive-negative compound amplitudes versus stripe size, ranged from 0.130 to 0.215. This acuity level is comparable to other inshore, shallow water marine species. PMID- 11913814 TI - Contact behaviour of tenent setae in attachment pads of the blowfly Calliphora vicina (Diptera, Calliphoridae). AB - To enable strong attachment forces between pad and substrata, a high proximity between contacting surfaces is required. One of the mechanisms, which can provide an intimate contact of solids, is a high flexibility of both materials. It has been previously presumed that setae of hairy attachment pads of insects are composed of flexible cuticle, and are able to replicate the surface profile. The aim of this work was to visualise the contact behaviour of the setae by freezing substitution technique to understand setal mechanics while adhering to a smooth surface. This approach revealed considerable differences in the area of the setal tips between contacting and non-contacting pulvilli. Based on the assumption that setae behave like a spring pushed by the tip, a spring constant of 1.31 N m(-1) was calculated from direct measurements of single setae by atomic force microscopy. In order to explain the relationship between the behaviour of the attachment setae at a microscale and leg movements, high-speed video recordings were made of walking flies. This data show that some proximal movement of the leg is present during contact formation with the substrate. PMID- 11913815 TI - Temperature dependence of cicada songs (Homoptera, Cicadoidea). AB - The songs of male Portuguese cicadas Tettigetta argentata, T. josei and Tympanistalna gastrica were recorded at five to seven temperatures within the range 24-38.5 degrees C. To investigate the temperature dependence of the neuromuscular apparatus involved in song production, different temporal elements of the calling songs were measured. We report a strong temperature dependence for the syllable and the echeme rates in T. josei and Ty. gastrica. This suggests that in these species the neuromuscular structures involved in the timbal cycle and in generating the echeme succession of the song are strongly temperature dependent. In T. argentata, the syllable rate was again significantly temperature dependent; the echeme rate, however, increased between 25.5 degrees C and 33.5 degrees C but decreased with the highest temperature. This indicates that at least in T. argentata two separate neuronal networks control both song parameters. Other temporal elements of the song with potential behavioural significance were also measured and found to be temperature dependent (e.g. echeme duration and interval). The possible implications for intraspecific communication are discussed. We also demonstrate that the temperature of these small cicadas is not significantly influenced by the muscle activity involved in song production. On the other hand, exposure to sunlight can be used by these cicadas to elevate their body temperature by more than 10 degrees C. PMID- 11913816 TI - Coding of a sexually dimorphic song feature by auditory interneurons of grasshoppers: the role of leading inhibition. AB - The shape of stimulus onset is a distinct feature of many acoustic communication signals. In some grasshopper species the steepness of amplitude rise of the pulses which comprise the song subunits is sexually dimorphic and a major criterion of sex recognition. Here, we describe potential mechanisms by which auditory interneurons could transmit the information on onset steepness from the metathoracic ganglion to the brain of the grasshopper. Since no single interneuron unequivocally encoded onset steepness, it appears that this information has to reside in the relative spike counts or the relative spike timing of a small group of ascending auditory interneurons. The decisive component of this mechanism seems to be the steepness-dependent leading inhibition displayed by two interneurons (AN3, AN4). The inhibition increased with increasing onset steepness, thus delayed the excitatory response, and in one interneuron even strongly reduced the spike count. Other ascending interneurons, whose responses were little affected by onset steepness, could serve as reference neurons (AN6, AN12). Thus, our results suggest that a comparison of both, spike count and first-spike timing within a small set of ascending interneurons could yield the information on signal onset steepness, that is on the sex of the sender. PMID- 11913817 TI - Motor program initiation and selection in crickets, with special reference to swimming and flying behavior. AB - An air puff stimulus to the cerci of a cricket (Gryllus bimaculatus) evokes flying when it is suspended in air, while the same stimulus evokes swimming when it is placed on the water surface. After bilateral dissection of the connectives between the suboesophageal and the prothoracic ganglia or between the brain and the suboesophageal ganglion, the air puff stimulus evokes flying even when the operated cricket is placed on the water surface. A touch stimulus on the body surface of crickets placed on the water surface elicits only flying when the connectives between suboesophageal and prothoracic ganglia are dissected, while the same stimulus elicits either swimming or flying when the connectives between the brain and the suboesophageal ganglion are dissected. These results suggest that certain neurons running through the ventral nerve cords between the brain and the suboesophageal ganglion or between the suboesophageal and the prothoracic ganglia play important but different roles in the initiation and/or switching of swimming and flying. In the suboesophageal ganglion, we physiologically and morphologically identified four types of "swimming initiating neurons". Depolarization of any one of these neurons resulted in synchronized activities of paired legs with a similar temporal sequence to that observed during swimming. PMID- 11913818 TI - Depth generalization from stereo to motion parallax in the owl. AB - Although many sources of three-dimensional information have been isolated and demonstrated to contribute independently, to depth vision in animal studies, it is not clear whether these distinct cues are perceived to be perceptually equivalent. Such ability is observed in humans and would seem to be advantageous for animals as well in coping with the often co-varying (or ambiguous) information about the layout of physical space. We introduce the expression primary-depth-cue equivalence to refer to the ability to perceive mutually consistent information about differences in depth from either stereopsis or motion-parallax. We found that owls trained to detect relative depth as a perceptual category (objects versus holes) when specified by binocular disparity alone (stereopsis), immediately transferred this discrimination to novel stimuli where the equivalent depth categories were available only through differences in motion information produced by head movements (observer-produced motion parallax). Motion-parallax discrimination did occur under monocular viewing conditions and reliable performance depended heavily on the amplitude of side-to side head movements. The presence of primary-depth-cue equivalence in the visual system of the owl provides further conformation of the hypothesis that neural systems evolved to detect differences in either disparity or motion information are likely to share similar processing mechanisms. PMID- 11913819 TI - Effects of anabolic implants of oestradiol alone or in combination with trenbolone acetate on the ultrastructure of mammary glands in female lambs regarding their interference in prolactin secretion. AB - The side-effects of anabolic steroid implants on mammary gland ultrastructure were evaluated in female lambs treated with oestradiol (n = 10) and with oestradiol plus trenbolone acetate (n = 10). Ten non-implanted lambs were used as controls. Apart from the ultrastructural study of the mammary gland, an assessment of the prolactin pituitary cell population was carried out by immunological methods. Our results showed that oestrogenic implants exert stimulating effects on mammary gland development, both by activating the synthesis process at mammary gland cell levels and by increasing prolactin pituitary production. Nevertheless, there was no evidence of secretory products in the lumen of the gland. Implants containing trenbolone acetate counteracted the mammary stimulus of oestrogens showing ultrastructural images of cell autolysis and necrosis. PMID- 11913820 TI - Study of the distribution of inflammatory cells in the sow endometrium: effect of intravenous administration of adrenocorticotropin hormone. AB - Seventeen multiparous cross-bred sows (Swedish Land-race x Swedish Yorkshire) were inseminated in their second oestrus after weaning and divided into two groups. One group (ACTH, n = 9) was given an intravenous injection of adrenocorticotropin hormone (ACTH) every 6 h commencing 4-8 h after ovulation, whereas another group (control, n = 8) was given saline solution at the same times. The sows were slaughtered 35-53 h after ovulation. Uterine samples, taken from the mesometrial side of the uterine horns immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue. The endometrium was then examined by light microscopy. There was no significant effect of the ACTH treatment on the distribution of lymphocytes and macrophages, but there was a tendency of an effect on the distribution of neutrophils (P = 0.1) in the sow endometrium. PMID- 11913821 TI - Prevention of scours in neonatal kids after oral administration of an organic acid solution. AB - The purpose of this study was to evaluate under field conditions, the efficacy of mild organic acid solutions in the prevention of neonatal kid diarrhoea. At a goat farm, two experimental groups of approximately 120 kids each were formed. The kids of the first group were not submitted to any treatment and served as negative controls, whereas the kids of the second group received a solution of organic acids (Euroacid 50-L; Eurotec, Waterloo, Belgium) which was administered orally on the first and second day of life. Groups were compared with regard to the incidence of diarrhoea, its duration, and the mortality of the kids. The results showed that, in comparison with the control group, the morbidity, the mortality and the case fatality of the treated kids was significantly reduced (P < 0.05). Furthermore, the duration of diarrhoea per sick kid was markedly reduced in the acidifier-treated group in comparison with the control animals (P < 0.05). Cultures of the diarrhoeic faeces from kids indicated that enterotoxigenic strains of Escherichia coli, and more specifically strains that were positive for K88 and K99 antigens, were present in the particular farm. It was concluded that the administration of organic acids can be a helpful means in controlling scours in neonatal kids. PMID- 11913822 TI - Expression of T helper 1 and T helper 2 cytokine mRNAs in freshly isolated peripheral blood mononuclear cells from dogs with atopic dermatitis. AB - Using reverse transcription-polymerase chain reaction (RT-PCR) and semi quantitative PCR techniques, mRNA expression for canine interferon (IFN)-gamma, interleukin (IL)-4, IL-5 and IL-10 in freshly isolated peripheral blood mononuclear cells (PBMCs) was examined in dogs with or without atopic dermatitis (AD). The expression of IFN-gamma mRNA in dogs with AD was lower than that in dogs without AD (healthy control). The expression of IL-5 mRNA was higher in dogs with AD than in control dogs, but there were no significant differences in IL-4 mRNA and IL-10 mRNA expression between the groups. The number of circulating eosinophils was higher in dogs with AD than in control dogs, although eosinophilia was found in only one dog with AD. These results suggest that there is a tendency for the PBMCs of atopic dogs to express a type 2 cytokine pattern that is similar to the pattern observed in human AD patients. PMID- 11913823 TI - Analgesic, sedative and haemodynamic effects of spinally administered romifidine in female goats. AB - The present study was designed to evaluate the analgesic, sedative and haemodynamic effects of spinally administered romifidine in goats. Ten female healthy goats weighing 14-18 kg were randomly divided into two groups, I and II, of five animals each. Romifidine was administered spinally at rates of 50 and 75 microg/kg body weight in the animals of groups I and II, respectively, into the lumbosacral space. The treatments were compared based on their effects on analgesia, sedation, ataxia, heart rate, respiratory rate, rectal temperature, mean arterial pressure, central venous pressure, electrocardiogram and haemato biochemical parameters. The objective parameters were analysed statistically using paired t-test and Duncan's multiple range test. Depth of analgesia was measured by recording the response to pin prick at different regions and was graded on a scale from 0 to 3. Moderate to complete analgesia was recorded at perineum and flank in both groups. Sedation was moderate in both groups. Ataxia was observed in all the animals but it was more pronounced in group II. Heart rate decreased significantly (P < 0.01) in both groups. A decrease in respiration rate was also recorded in both groups but it was more significant (P < 0.01) and for longer duration in group II as compared to group I. A slight increase in rectal temperature was also observed in both groups. Mean arterial pressure decreased and central venous pressure increased significantly (P < 0.01) in both groups but changes were more pronounced in group II. Electrocardiogram changes in group I included bradycardia, increased QT interval and increased or biphasic T wave but in animals of group II, in addition to these changes, occasional sinus dysrhythmia, increased PR interval and second-degree heart block were also recorded. Haemoglobin and packed cell volume decreased non-significantly in both groups. A significant (P < 0.01) increase in blood glucose and non-significant changes in plasma proteins, urea nitrogen and creatinine were recorded in both groups. The results of the study revealed that romifidine at the rate of 50 microg/kg could produce moderate to complete analgesia of perineum and flank after spinal administration into the lumbosacral space in goats. The analgesia could not be enhanced further by increasing the dose of romifidine up to 75 microg/kg, however, ataxia and cardiopulmonary and haemodynamic side-effects became more apparent. PMID- 11913824 TI - The effect of short-duration, high-intensity electromagnetic pulses on fresh ulnar fractures in rats. AB - Pulsed electromagnetic fields (PEMFs) have been found to be beneficial to a wide variety of biological phenomena. In particular, PEMFs have been shown to be useful in the promotion of healing of ununited fractures. Conflicting information exists regarding the benefit of using PEMFs to accelerate the healing of fresh fractures. This paper reports on the evaluation of the effect of a new PEMF generator (PAP IMI) on the healing of fresh ulnar fractures in rats. This device is unique by virtue of the extremely high power output of each of the pulses it generates. Ulnar fractures were created in rats by using a bone cutter, thus producing a 2-3 mm bone defect. Rats were then randomly divided into treatment and control groups. The treatment group underwent periodic treatments with the PAP IMI, and the control group received no treatment. Radiographs of rats from both groups were taken at 1-week intervals. Histological evaluation was performed at the end of the study. Radiographic and histopathological evaluations were scored, and scores were used to assess both rate and quality of healing. The radiographic results demonstrated gradual bridging callus formation in both control and treatment groups, however, the healing process was faster in rats that were not treated by PEMF. Histological evaluation demonstrated that the fibrous content of the callus in rats belonging to the treatment group was significantly higher than that in rats belonging to the control group. The results of this study do not support the claim that PEMF generated by the PAP-IMI stimulate osteogenesis and bone healing after the creation of fresh ulnar fractures in rats. PMID- 11913825 TI - In vitro studies of a photo-oxidized bovine articular cartilage. AB - Bovine articular cartilage was photo-oxidized and cultured with native articular bovine cartilage and synovial membrane to study the interaction between these tissues mimicking the physiological situation in the joint. The photo-oxidation was applied as a pretreatment of cartilage for future use in cartilage resurfacing procedures in joints. Properties of the transplant were assessed by testing the production of local mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), and neutral metalloproteinase activities under normal conditions and after stimulation with various stimulants representative of inflammatory changes in pathophysiological conditions. Unlike normal cartilage photo-oxidized cartilage did not release significant amounts of NO and PGE2 and showed less gelatinolytic and caseinolytic activity compared to native bovine articular cartilage. Enzyme activity of the combined cultures was at a level intermediate between that of photo-oxidized cartilage and native cartilage cultures alone. In contrast to normal cartilage, living chondrocytes were not visible in photo-oxidized cartilage using live/dead staining. These results indicate, that the photo-oxidized cartilage may have a beneficial effect on adjacent native host cartilage and therefore be a suitable transplant for use in in vivo experiments. PMID- 11913826 TI - Necrotizing cerebellitis due to Neospora caninum infection in an old dog. AB - A severe, necrotizing, non-suppurative inflammation of the cerebellum associated with Neospora caninum infection was identified in a 14-year-old male Labrador Retriever. On presentation, clinical signs included mild depression and head tremor, marked ataxia of both thoracic and pelvic limbs, and abnormal postural reactions. In the central nervous system, inflammatory lesions were mainly restricted to the cerebellar leptomeninges and cerebellar cortex, which appeared necrotic and atrophic. Protozoal organisms were positively stained with an anti N. caninum antibody in an immunohistochemical procedure. PMID- 11913827 TI - Functional changes in isolated guinea-pig papillary muscle induced by monensin and digoxin. AB - The effects of digoxin and monensin on contraction force (CF), initial contraction velocity (ICV), average contraction velocity (ACV), initial relaxation velocity (IRV) and stimulus to response time (ST) in 'fatigued' (tired) and 'non-fatigued' (fresh) guinea-pig papillary muscles were investigated. 'Fatigued' muscles had lost 30% of their original CF with the elapse of time before they were treated. The 5 h of measurement were divided into five periods (T0 was equilibration, T1, T2, T3 and T4 were, respectively, 1, 2, 3 and 4 h after drug administration). It was found that both monensin and digoxin increased the CF, ICV and ACV at T1 and increased the IRV at T2. Digoxin lost its effect with the elapse of time while monensin did not. Digoxin also decreased the ST at T2, T3 and T4. However, monensin did not change the ST. It was also found that 'fatigued' and 'non-fatigued' guinea-pig papillary muscles did not respond to the drug treatment differently. It was concluded that the initial effects of these two drugs on guinea-pig papillary muscles are similar regarding contractility but in time digoxin loses its effect while monensin does not. PMID- 11913828 TI - Serum lipid profile in Iranian fat-tailed sheep in late pregnancy, at parturition and during the post-parturition period. AB - Blood samples were obtained from 12 Iranian fat-tailed sheep during 7 weeks pre partum, at parturition and 7 weeks post-partum. The lipids measured were cholesterol, triglyceride, total lipid, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL)-cholesterol, and very low-density lipoprotein (VLDL)-cholesterol. The concentrations of cholesterol, triglyceride, HDL-cholesterol and VLDL-cholesterol during the 7 weeks pre-partum, at parturition and the 7 weeks post-partum were significantly different (P < 0.05). One week before parturition, the concentrations of cholesterol, triglyceride, HDL cholesterol and VLDL-cholesterol were higher (P < 0.05) than at other periods. The lowest concentrations of these parameters were observed 2-3 weeks after parturition. In this study, significant positive correlations were observed between the time of sampling (pre-partum, parturition and post-partum) and serum cholesterol (r = 0.22; P < 0.01) and HDL-cholesterol (r = 0.25; P < 0.01). PMID- 11913829 TI - The moderating roles of personal and social resources on the relationship between dual expectations (for instrumentality and expressiveness) and well-being. AB - The authors examined the roles of self-esteem and perceived social support in moderating the relationship between dual social expectations (instrumentality and expressivity) and well-being. Participants were 166 female and 87 male students in an urban community in the midwestern United States. After the authors controlled for the main effects of instrumental and expressive expectations, social support, as predicted, moderated the relationship between dual expectations and well-being: With higher levels of social support, higher levels of dual expectations were associated with higher levels of well-being; with lower levels of social support, higher levels of dual expectations were associated with lower levels of well-being. Contrary to predictions, however, self-esteem did not moderate the relation between dual expectations and well-being. The discussion focuses on the importance of social resources in enhancing the potential benefits of dual expectations in interpersonal contexts. PMID- 11913830 TI - The interactive effects of affective demeanor, cognitive processes, and perspective-taking focus on helping behavior. AB - The author manipulated affective demeanor (positive or negative) and cognitive processes (positive or negative) displayed by a target person, along with the perspective-taking focus (affect or cognitions) of participants, to assess the unique and interactive effects of those variables on the participants' helping behavior, operationalized as time volunteered to help other students. An ethnically diverse sample (N = 109) of U.S. working adults (mean age = 31.56 years, SD = 8.21) viewed a videotape of a female target talking about returning to college. Participants adopting an affective perspective-taking focus volunteered more time than did those who adopted a cognitive perspective-taking focus. Also, a significant interaction between participants' perspective-taking focus and target's affective demeanor revealed that participants who focused on the target's feelings and who viewed a warm, cheerful target volunteered more time than did the other groups. Moreover, a significant interaction between participants' perspective-taking focus and target's cognitive processes revealed that the participants who focused on the target's feelings and who viewed a confused and unfocused target volunteered more time than did the other groups. The author also discusses the relationship between empathy, personal distress, and helping. PMID- 11913831 TI - Is emotional intelligence an advantage? An exploration of the impact of emotional and general intelligence on individual performance. AB - Emotional intelligence is an increasingly popular consulting tool. According to popular opinion and work-place testimonials, emotional intelligence increases performance and productivity; however, there has been a general lack of independent, systematic analysis substantiating that claim. The authors investigated whether emotional intelligence would account for increases in individual cognitive-based performance over and above the level attributable to traditional general intelligence. The authors measured emotional intelligence with the Multifactor Emotional Intelligence Scale (MEIS; J. D. Mayer, P. Salovey, & D. R. Caruso, 1997). As measured by the MEIS, overall emotional intelligence is a composite of the 3 distinct emotional reasoning abilities: perceiving, understanding, and regulating emotions (J. D. Mayer & P. Salovey, 1997). Although further psychometric analysis of the MEIS is warranted, the authors found that overall emotional intelligence, emotional perception, and emotional regulation uniquely explained individual cognitive-based performance over and beyond the level attributable to general intelligence. PMID- 11913832 TI - Physical attractiveness, social connectedness, and individuality: an autophotographic study. AB - In a sample of 125 college students, the author tested the hypothesis that, relative to less attractive peers, the physically attractive participants would depict greater social connectedness in self-descriptive photo essays. To document discriminant validity, the author also hypothesized that the correlation between physical-attractiveness ratings and the richness (i.e., individuality ratings) of photo essays would be near zero. The results confirmed both predictions and supported the convergent and discriminant validity of information from autophotographic essays. Thus, the attractive participants depicted themselves as very socially connected; however, they were neither more conventional-superficial nor more individualistic-creative than their less attractive peers in their photo essays. PMID- 11913833 TI - Justice and organizational citizenship behavior intentions: fair rewards versus fair treatment. AB - In a sample of 114 employees from various industries, organizations, and positions, the likelihood of organizational citizenship behaviors (OCB) increased when employee perceptions of fair treatment by supervisors became more positive. Perceptions of fair rewards and fair formal procedures were not predictors of OCB intentions. After the authors controlled for established patterns of OCB and demographic characteristics, interactional justice perceptions were significantly related to the intention of performing specific organizationally beneficial activities. PMID- 11913834 TI - Traditional and modern characteristics across the generations: similarities and discrepancies. AB - The authors examined, through an analysis of self-construals and beliefs about social interaction, the traditional and modern orientations of younger and older generations of Taiwanese people. The authors surveyed 169 pairs of parent-child dyads with a battery of structured questionnaires. Within-subjects analyses revealed high generational correlations as well as discrepancies in both traditional and modern characteristics. Between-subjects analyses further indicated that individual traditional and modern characteristics were affected by age and gender and by their interactions. The authors discuss (a) the pattern of generational similarities and discrepancies in the framework of culture stability and change and (b) the coexistence of traditional and modern characteristics in a rapidly changing society. PMID- 11913835 TI - Work-group characteristics and performance in collectivistic and individualistic cultures. AB - The authors conducted a cross-cultural longitudinal investigation of the effects of culture (individualism-collectivism dichotomy) on group characteristics (functional heterogeneity, preference for teamwork, group potency, outcome expectation) and on performance of 83 work groups performing 2 decision-making tasks over a 15-week period. The individualists (U.S. students) reported higher levels of functional heterogeneity and group potency and attained higher levels of group performance than did the collectivists (Korean students). In addition, culture and time interacted to influence ratings of group potency and outcome expectation. The difference in ratings of group potency between individualists and collectivists increased over time. Outcome expectation was greater among the collectivists in Time 1 and among the individualists in Time 2. The authors discuss implications for future cross-cultural group research and international management. PMID- 11913836 TI - How a group goal may reduce social matching in group performance: shifts in standards for determining a fair contribution of effort. AB - The authors investigated whether the presence of a specific group goal would reduce social matching (i.e., matching one's own performance to the performance expected from others) by serving as an alternative standard. As predicted, when there was no specific goal, the participants matched their own performance to the performance expected from other group members. When there was a specific group goal, the women no longer engaged in social matching, although that effect did not emerge among the men. Instead, the women's mean personal performance was close to the performance level representing an equal share of the group goal. Moreover, the participants' perceptions of a fair contribution mediated the performance of the men and the women, both in the presence and in the absence of a goal. PMID- 11913838 TI - Avoiding conflicts--an editorial update. PMID- 11913839 TI - President fulfills commitment to doubling NIH funding. PMID- 11913837 TI - Personal goals and depression among Vietnamese American and European American young adults: a mediational analysis. AB - The authors report preliminary findings supporting the utility of the self concordance model (K. M. Sheldon & A. J. Elliot, 1999) as an alternative approach to studying depression among Vietnamese American (N = 121) and European American (N = 155) college students. The participants completed measures of personal goals, goal self-concordance, and depression. Compared with the European American participants, the Vietnamese American participants reported higher levels of depression and lower levels of goal self-concordance. According to mediational analyses, ethnicity no longer accounted for significant variance in depression after the authors statistically controlled for goal self-concordance. PMID- 11913840 TI - Explanation of the lens paradox. PMID- 11913841 TI - A survey of treatment modalities for convergence insufficiency. AB - BACKGROUND: Convergence insufficiency (CI) is a common and distinct binocular vision disorder. However, there is a lack of consensus regarding the treatment most appropriate for Cl. Possible treatment modalities include base-in prism, pencil pushup therapy (PPT), reading glasses, home-based vision therapy/orthoptics (HBVT), and office-based vision therapy/orthoptics (OBVT). The purpose of this study was to investigate the care process for Cl by surveying eyecare professionals regarding the most common treatment modalities used by both optometrists and ophthalmologists across the United States. METHODS: Surveys requesting doctors to indicate which treatment(s) they prescribed and believed to be most effective for symptomatic CI patients were mailed to 863 optometrists and 863 ophthalmologists in the United States. RESULTS: Fifty-eight percent of the optometrists responded to the survey; the most common treatment prescribed was PPT (36%) followed by HBVT (22%) and OBVT (16%). For the ophthalmologists (who had a 23% response rate), the most common treatment prescribed was PPT (50%) followed by HBVT (21 %) and base-in prism (10%). CONCLUSIONS: This survey suggests that most eyecare practitioners prescribe PPT as the initial treatment for CI. PMID- 11913842 TI - The proximity-fixation-disparity curve and the preferred viewing distance at a visual display as an indicator of near vision fatigue. AB - PURPOSE: This laboratory study investigates the relation between measures of fixation disparity (FD) (and other optometric measures) and near vision fatigue at a computer workstation. METHODS: Young adult subjects with normal binocular vision performed three blocks of a visual task of 30 min each. In Block A, the viewing distance was 100 cm, as a reference without near vision. In Block B, the viewing distance of 50 cm induced a defined near vision load. In Block C, subjects were free to choose a comfortable viewing distance. This preferred viewing distance was used as an indicator of near vision fatigue because subjects adopting longer viewing distances in Block C had more near vision fatigue at 50 cm in Block B. RESULTS: Subjects with preferred viewing distances longer than average (63 cm) had steeper slopes of FD as a function of viewing distance (100 30 cm), as shown by discriminant analyses. CONCLUSIONS: Thus, this steep proximity-FD curve indicates a weak disparity vergence system that may cause near vision fatigue. This may explain why some young adults prefer longer viewing distances at the computer workstation. PMID- 11913843 TI - Maximum disparity with acuvue bifocal contact lenses with changes in illumination. AB - PURPOSE: To assess the visual performance of Acuvue bifocal contact lenses at different illumination levels using a stereoscopic parameter, the maximum disparity. We compare the results with those for progressive spectacle lenses. METHODS: We used a modified Wheatstone stereoscope and random-dot stereograms (RDS) as tests. The maximum disparity was measured for each observer at different luminance levels. RESULTS: Maximum disparities did not show statistically significant differences for a wide range of luminance levels with contact lenses. The maximum disparities found when some contact-lens users wore progressive spectacle lenses were similar. CONCLUSIONS: The results show that the Acuvue bifocal lenses offer good performance for a near visual task, in this case the maximum disparity, for changes in illumination and are similar to that with progressive spectacle lenses. PMID- 11913845 TI - A quantitative analysis of sutural contributions to variability in back vertex distance and transmittance in rabbit lenses as a function of development, growth, and age. AB - PURPOSE: To correlate specific parameters of lens structure (anterior and posterior suture branch length and planar area) with variability in back vertex distance (BVD) and scatter in rabbit lenses as a function of development, growth, and age. METHODS: Lenses from juvenile (n = 9), adult (n = 9), and aged (n = 10) New Zealand White rabbits were utilized in this study. After sacrifice, lens suture patterns were photographed using a stereo surgical dissecting microscope. Within 5 min of sacrifice, average BVD, variability in BVD, and scatter were assessed with a Scantox In Vitro Assay System. Laser beams were passed incrementally along anterior and posterior suture planes through right eye (oculus dexter, OD) lenses, and between suture planes through left eye (oculus sinister, OS) lenses. After fixation, lens axial dimensions and suture branch lengths were assessed and used to create scaled, 3-dimensional computer assisted drawings (3-D CADs) depicting gross lens shape and sutural changes throughout life. RESULTS: Whereas average BVD only increased significantly as a function of growth, variability in BVD only increased significantly as a function of aging. However, anterior sutures exerted a greater influence on variability of BVD than posterior sutures throughout growth and aging. This difference is consistent with anterior suture branches being longer, or extending farther peripherally, than posterior sutures. Scatter was essentially unchanged between juvenile and adult lenses but significantly increased in aged lenses. Notably, posterior sutures effected a greater age-related increase in scatter than anterior sutures. This difference was consistent with the formation of numerous small, posterior subbranches and subplanes later in life. Structural analysis also suggested that asymmetric age-related lens compaction had occurred, predominantly affecting posterior lens dimensions. CONCLUSIONS: Lens sutures significantly influence average BVD throughout development and growth, and variability in BVD throughout aging. In addition, even though the rabbit lenses appeared transparent throughout growth and aging, unequal length and area of anterior vs. posterior suture branches and planes respectively, as well as a greater degree of age-related posterior lens compaction, were factors contributing to increased scatter. PMID- 11913844 TI - Overwear of contact lenses: increased severity of clinical signs as a function of protein adsorption. AB - This study compared the clinical behavior of disposable and frequent replacement Acuvue and 1-Day contact lenses. Each type of lens was worn on one eye according to the schedule recommended by the manufacturer, and on the other eye for a longer period of time, up to 30 days in length. Both type of lenses were prescribed on a daily-wear basis. The amount of protein collected from the lenses was measured using two spectrophotometric protein assays. Visual acuity and comfort, along with several other clinical signs, were classified according to Cornea and Contact Lens Research Unit (CCLRU) scales, and possible associations between each of these signs and the amount of protein extracted from the lenses was tested. A comparison between the lens worn on the compliant eye with the lens worn on the noncompliant eye allowed us to measure the impact of overwear on ocular health and subjective clinical findings. After four months of study, the overwear of Acuvue and 1-Day lenses significantly increased the amount of protein bound on the contact lenses, as well as the severity of upper conjunctival papillae, upper lid conjunctival hyperemia, and limbal congestion. Even if reduced values for visual acuity and noninvasive break-up time (NIBUT) were identified, these variations were not found to be statistically significant. The clinical implications of this study would allow a practitioner to identify, according to the variations in several clinical signs, a patient who overwears contact lenses, so that action may be taken to reduce possible deleterious effects on ocular health. PMID- 11913846 TI - Partner orientation and speaker's knowledge as conflicting parameters in language production. AB - Two parameters of language production, the partner model and the mentally represented knowledge about a spatial constellation, are investigated with respect to their influence on spatial reference. At issue is whether the verbally expressed point of view in route directions is primarily influenced by the needs of the partner or by the underlying mental representation of the speaker stemming from his or her own experience with an object. Two experiments in which participants (N = 90) were asked to produce a set of route directions are reported. The experimental situation was such that the point of view of the speaker did not correspond to the point of view of the partner. The results show that more participants localize from their own point of view than from the point of view of the partner. Discussion centers on the fact that speakers do not always behave in a truly partner-oriented manner. PMID- 11913847 TI - Competition between primary and non-primary relations during sentence comprehension. AB - The four eye-tracking experiments reported examined the way in which Adjunct Predicates (APs) located at the beginning of French sentences of the type "Tired (feminine/masculine) of calling the woman (s/he) left the room" are interpreted and interact with syntactic parsing strategies. The results suggest that the first NP (the woman) was initially interpreted as the potential AP gender controller. Moreover, in the case of gender agreement (the woman is the one who is tired) the syntactic status of the first NP (either the object of the preceding verb or the subject of the main verb) apparently remained ambiguous until the main verb was reached. The implications of these results for Frazier and Clifton's (1996) Construal Theory are discussed. PMID- 11913849 TI - Essential and perceptual attributes of words in reflective and on-line processing. AB - Access to essential and perceptual attributes of word meanings was investigated in a series of tasks. First, perceptual attributes of a number of natural kind concepts were determined by asking participants to generate the most salient features of those concepts; they were asked to assess the importance of both thosefeatures and features that fit criteria for essential attributes. Attributes fitting the criteria for essential attributes were found to be judged more important than perceptual attributes in identifying an object and to have more connections to other attributes of the concept. Semantic priming for both types offeatures was then investigated. Prime-target pairs consisting of the natural kind concept term and the perceptual or essential attribute were presented for lexical decision (Experiments 1a and 1b) or naming (Experiments 2a and 2b). Interstimulus intervals were either 0 or 200 ms. Both types of features were primed at a 200 ms interstimulus interval but only in the lexical decision task. PMID- 11913848 TI - The pronoun bias effect strikes again: a reply to Pynte and Colonna. AB - The gender of an initial adjectival phase (AP) influences the analysis of a phrase which is a potential main clause subject in French. Pynte and Colonna (this issue) establish this generalization but analyze it without considering critical aspects of sentence structure, specifically, the fact that the AP has a null pronominal subject. If grammars contain a violable constraint requiring coreference between the pronominal subject of an initial subordinate phrase and the subject of the main clause, then in Pynte and Colonna's study perceivers must choose on-line between an analysis which satisfies the same subject constraint and one which doesn't. From this perspective, it is expected that the subject analysis of the ambiguous phrase predominates. Further, on this view, the preference fits with independent generalizations about language such as the existence of switch-reference languages which obligatorily mark the coreference relation between the initial subordinate subject and the main clause subject. It also fits with already established facts about language processing, such as Cowart and Cairns' (1987) Pronoun Bias Effect. PMID- 11913850 TI - Two routes to grammatical gender: evidence from Hebrew. AB - Access to grammatical gender in Hebrew was examined using gender decisions and grammaticality judgments. In gender decisions, and in rejecting agreement violations, the role of gender-marking regularity was robust; irregular feminine nouns were classified more slowly and produced more errors than did regular masculine nouns. These same exception nouns, however, produced radically different results when they appeared as part of a grammatical phrase. In this context, irregular nouns were processed as quickly and accurately as were regular nouns. These findings are interpreted within a model containing two routes to grammatical gender: one that involves an abstract gender node, and another that is form-based and is assumed to play a greater role in recovery from agreement errors. We also argue that there are cross-linguistic variations in the availability and speed of the form-based route to gender. Finally, we caution that under some circumstances grammatical gender may also be retrieved through a variety of heuristics that do not necessarily reflect the normal mechanisms of access to gender. PMID- 11913851 TI - Filler + infinitive and pre- and protomorphology demarcation in a French acquisition corpus. AB - This paper presents a case study on the acquisition of grammatical morphemes via fillers, i.e., underspecified place holders, with particular focus on early structures made up of a filler followed by an infinitive. The path leading from fillers to French semi-auxiliaries and subject clitics is analyzed within the framework of Natural Morphology and constructivism which assumes that grammatical modules are not innate but are constructed by children. The evolution of fillers in the corpus studied is described as a grammaticization process of form and meaning through successive linguistic dissociations. Emphasis is put on the functional polyvalence of fillers and on their relation to the main phases in the construction of grammar. PMID- 11913852 TI - The anatomy of the job-generation issue and its impact on health insurance policy. AB - Small business does not create most jobs in the United States, but, as the author argues, small business has successfully used this myth to counter demands that it should provide health insurance for its workers. What is at stake in this job generation claim is no less than the economic well-being of millions of Americans. PMID- 11913853 TI - Pathbreaking Congressional Budget Office study shows dramatic increases in income disparities in the 1980s and 1990s: an analysis of the CBO data. AB - Researchers at the Center on Budget and Policy Priorities analyze the data presented in a Congressional Budget Office study that includes the best data any agency or institution has compiled on income and tax trends in recent decades. The CBO report shows that the average after-tax income of the richest 1 percent of Americans grew by $414,000 between 1979 and 1997 (after adjusting for inflation) while average after-tax income fell $100 for the poorest 20 percent of Americans and grew a modest $3,400 for those in the middle of the income spectrum. In percentage terms, after-tax income grew an average of 157 percent over this period for the top 1 percent, rose a modest 10 percent for the middle 20 percent, and was effectively unchanged for those in the bottom fifth. Income gaps between rich and poor and between the rich and the middle class widened in the 1980s and 1990s and reached their widest point on record in 1997. Even before enactment of the 2001 federal tax cuts, the percentage of income Americans pay in federal taxes has declined since 1979 for every income group. By one key measure, the percentage of income paid in federal taxes fell the most for those with the highest incomes. PMID- 11913855 TI - Accident insurance, sickness, and science: New Zealand's no-fault system. AB - This article explores the process of seeking compensation for occupational illness under a no-fault accident insurance scheme. The author uses two case studies--firefighters who attended a fire at a chemical storage depot and timbermill workers who worked with pentachlorophenol--to illustrate how science can be used to deny compensation to sick and dying workers. The results of the studies suggest that a no-fault accident compensation scheme, considered to be a victory for workers, offers no guarantee of just outcomes for working people. And science can be co-opted and used to support business and state interests against workers; this ideological support is increasingly hidden behind the development of "objective" systems of assessing compensation claims. PMID- 11913854 TI - Oral health status and its inequality among education groups: comparing seven international study sites. AB - This study compares oral health status and its inequality among education groups across seven study sites in five countries: Erfurt, Germany; Lodz, Poland; Yamanashi, Japan; New Zealand; and Baltimore and the Lakota and Navajo Indian Health Service sites in the United States. The data, from the International Collaborative Study of Oral Health Outcomes, were collected through personal interviews and clinical examinations. The research group measured the study sites' overall oral health, examining the percentage of the population with five or more missing and two or more decayed teeth. The group also assessed the magnitude of inequality among education groups by using indices of excess morbidity. Baltimore had the lowest percentage (10.8 percent) of decayed teeth and second lowest percentage (17.3 percent) of missing teeth, but the greatest indices of excess morbidity (79.2 percent for missing, 73.1 percent for decayed). Lodz, by contrast, had the worst overall dentition status (75.3 percent for missing, 70.3 percent for decayed) but the lowest inequality indices (10.6 percent for missing, 13.8 percent for decayed). This study demonstrates the need for policymakers in the study countries to consider not only overall levels but also the distribution of oral health, and it presents various challenges for oral health professionals in designing and implementing oral health programs. PMID- 11913856 TI - Role of trade unions in workplace health promotion. AB - Since the 19th century, workers have organized in trade unions and parties to strengthen their efforts at improving workplace health and safety, job conditions, working hours, wages, job contracts, and social security. Cooperation between workers and their organizations and professionals has been instrumental in improving regulation and legislation affecting workers' health. The authors give examples of participatory research in occupational health in Denmark and Finland. The social context of workplace health promotion, particularly the role of unions and workers' safety representatives, is described in an international feasibility study. Health promotion is rife with fundamental political, socioeconomic, philosophical, ethical, gender- and ethnicity-related, psychological, and biological problems. Analysis of power and context is crucial, focusing on political systems nationally, regionally, and globally. The authors advocate defending and supporting workers and their trade unions and strengthening their influence on workplace health promotion. In the face of rapid capitalist globalization, unions represent a barricade in defense of workers' health and safety. Health promoters and related professionals are encouraged to support trade unions in their efforts to promote health for workers and other less privileged groups. PMID- 11913857 TI - World Health Report 2000 "Financial fairness indicator": useful compass or crystal ball? AB - The World Health Report 2000 generated a huge amount of controversy when it set out to rank the performance of national health systems using data, statistical measures, and an explanatory rationale that were neither well understood nor broadly accepted. This article demystifies the conceptual and empirical underpinnings of the report's "financial fairness index," which resulted in country rankings that often seem counterintuitive. The author concludes that the index is seriously flawed, that rankings produced by the index should not be used, and that future WHO reports should avoid imputing financial fairness scores for countries that do not have real data. PMID- 11913858 TI - The Glaxo Award: is the American Public Health Association moving toward corporate governance? PMID- 11913859 TI - Millions of mothers lack health insurance coverage in the United States. Most uninsured mothers lack access both to employer-based coverage and to publicly subsidized health insurance. AB - Some 5.9 million American mothers caring for young or school-aged children lack health insurance. Although nearly nine in ten uninsured mothers are members of working families, most lack access to affordable coverage through their job or a spouse's job. Most are ineligible for publicly subsidized coverage unless their incomes are far below the poverty line. The millions of uninsured mothers are at high risk of going without needed preventive and primary care. If they become seriously ill, their families can face the prospect of a financial crisis. The nation has made significant progress in extending health care coverage to children in low-income families through Medicaid and the State Children's Health Insurance Program (SCHIP), but no comparable effort has been made to insure the mothers of these children. A few states have started to address the problem by transforming their SCHIPs into family-based programs that also cover low-income parents. Bipartisan legislation under consideration, known as FamilyCare, would encourage this trend by providing more federal funding to states that could be used to extend health insurance to the parents of children already covered by publicly funded programs. PMID- 11913862 TI - Are simple comparative studies always simple? The role of t-tests in comparative analytical experiments. PMID- 11913865 TI - Speciation of seleno compounds in yeast aqueous extracts by three-dimensional liquid chromatography with inductively coupled plasma mass spectrometric and electrospray mass spectrometric detection. AB - A three-dimensional liquid chromatographic purification protocol based on sequential size-exclusion, anion-exchange and cation-exchange separation mechanisms was developed for the mapping of seleno compounds in aqueous yeast extracts. The method allowed the demonstration of the presence of more than 30 different seleno compounds. Semi-preparative size-exclusion and anion-exchange chromatography were optimized for maximum resolution using electrospray compatible buffers in order to purify the compounds for mass spectrometric analysis. Molecular masses were attributed to many of the compounds on the basis of the selenium isotopic pattern in the electrospray mass spectra and of the collision-induced fragmentation patterns. Limitations preventing the ultimate identification of the selenium species detected are discussed. PMID- 11913867 TI - Amperometric microcells for alkaline phosphatase assay. AB - To develop simple electrochemical immunoassays, a screen printed amperometric microcell with graphite working and Ag/AgCl reference electrodes was tested for the determination of alkaline phosphatase enzyme (ALP) and anti-humanIgG conjugated ALP (alpha-hIgG-ALP) activity in 5-10 microl samples. To ensure reproducible, steady state conditions, the working electrode surface was coated with mass-transport controlling hydrogel layer. The kinetic response curves of the hydrogel coated electrodes were linear. In addition, the hydrogel layer reduced the nonspecific adsorption of the alpha-hIgG-ALP conjugate on the working electrode surface. The measurements were made in the range of 2 divided by 4000 mU ml(-1) enzyme activities using ascorbic acid 2-phosphate (AAP) as the enzyme substrate. AAP is commercially available, non-toxic and has excellent stability. The sensitivity of the determinations was about 71% of the sensitivity which could be achieved using p-aminophenylphosphate (PAPP), a not easily accessible and unstable enzyme substrate. The experimentally determined kinetic parameters of the ALP enzyme catalyzed reactions were the same with the bare and hydrogel layer coated electrodes. PMID- 11913873 TI - Metabolic trajectory characterisation of xenobiotic-induced hepatotoxic lesions using statistical batch processing of NMR data. AB - Multivariate statistical batch processing (BP) analysis of 1H NMR urine spectra was employed to establish time-dependent metabolic variations in animals treated with the model hepatotoxin, alpha-naphthylisothiocyanate (ANIT). ANIT (100 mg kg( 1)) was administered orally to rats (n = 5) and urine samples were collected from dosed and matching control rats at time-points up to 168 h post-dose. Urine samples were measured via 1H NMR spectroscopy and partial least squares (PLS) based batch processing analysis was used to interpret the spectral data, treating each rat as an individual batch comprising a series of timed urine samples. A model defining the mean urine profile over the 7 day study period was established, together with model confidence limits (+/-3 standard deviation), for the control group. Samples obtained from ANIT treated animals were evaluated using the control model. Time-dependent deviations from the control model were evident in all ANIT treated animals consisting of glycosuria, bile aciduria, an initial decrease in taurine levels followed by taurinuria and a reduction of tricarboxylic acid cycle intermediate excretion. BP provided an efficient means of visualising the biochemical response to ANIT in terms of both inter-animal variation and net variation in metabolite excretion profiles. BP also allowed multivariate statistical limits for normality to be established and provided a template for defining the sequence of time-dependent metabolic consequences of toxicity in NMR based metabonomic studies. PMID- 11913874 TI - Assessment of silver and gold substrates for the detection of amphetamine sulfate by surface enhanced Raman scattering (SERS). AB - Methods of detection of amphetamine sulfate using surface enhanced Raman scattering (SERS) from colloidal suspensions and vapour deposited films of both silver and gold are compared. Different aggregating agents are required to produce effective SERS from silver and gold colloidal suspensions. Gold colloid and vapour deposited gold films give weaker scattering than the equivalent silver substrates when high concentrations of drug are analysed but they also give lower detection limits, suggesting a smaller surface enhancement but stronger surface adsorption. A 10(-5) mol dm(-3) solution (the final concentration after addition of colloid was 10(-6) mol dm(-3)) of amphetamine sulfate was detected from gold colloid with an RSD of 5.4%. 25 microl of the same solution could be detected on a roughened gold film. The intensities of the spectra varied across the film surface resulting in relatively high RSDs. The precision was improved by averaging the scattering from several points on the surface. An attempt to improve the detection limit and precision by concentrating a suspension of gold colloid and amphetamine sulfate in aluminium wells did not give effective quantitation. Thus, positive identification and semi-quantitative estimation of amphetamine sulfate can be made quickly and easily using SERS from suspended gold colloid with the appropriate aggregating agents. PMID- 11913879 TI - Efavirenz and retinal toxicity. PMID- 11913880 TI - Benign idiopathic haemorrhagic retinopathy. PMID- 11913881 TI - 'One-stop' cataract surgery. PMID- 11913882 TI - Primary orbital melanoma masquerading as vascular anomalies. AB - PURPOSE: To review two cases of primary orbital melanoma presenting like orbital vascular anomalies. METHODS: Retrospective review of clinical presentation, treatment, radiology and pathology for two patients under the care of the Orbital Clinic at Moorfields Eye Hospital. RESULTS: Both lesions presented with the appearance and behaviour of vascular anomalies. In one case, a spindle cell melanoma appeared to be a low flow vascular anomaly with a loculated secondary haemorrhage and, in the other case, a melanoma of soft parts was considered to be an arteriovenous malformation and responded partially to embolisation. CONCLUSION: Primary malignant melanoma may present as a secondary vascular lesion of the orbit and this very rare tumour should be considered in the differential diagnosis of any vascular anomaly. PMID- 11913883 TI - Red and/or blonde hair association with pigmentary glaucoma in Israel. AB - PURPOSE: To assess the prevalence of red and/or blonde hair in a defined pigmentary glaucoma (PG) population and to compare their clinical findings with those of PG patients with black or brown hair. METHODS: Hair color was studied in 35 consecutive PG patients and 35 consecutive primary open angle glaucoma (POAG) patients who served as controls. Patients were classified into red and/or blonde hair group or into black or brown hair group. Clinical characteristics were contrasted between PG patients of red and/or blonde hair with PG patients of black or brown hair. RESULTS: Of the 35 PG patients, 19 (54.3%) had red and/or blonde hair and 16 (45.7%) had black or brown hair. Of the 35 POAG patients, two (5.8%) had red and/or blonde hair and 33 (94.2%) had black or brown hair. This difference in prevalence of hair color is highly significant (P < 0.001; chi2). Clinical characteristics of the PG patients (age at initial diagnosis, gender, positive family history for glaucoma, initial IOP, refraction, severity of optic nerve damage and prevalence of retinal detachment) were similar in the red and/or blonde hair and the black or brown hair groups. CONCLUSIONS: A very high prevalence of red and/or blonde hair was found among PG patients in Israel. Should supportive evidence for this association accumulate subsequent to this report, screening of people with these hair colors for PG may be justified. PMID- 11913884 TI - Nd:YAG vitreolysis and pars plana vitrectomy: surgical treatment for vitreous floaters. AB - PURPOSE OF STUDY: To determine the efficacy of Nd:YAG vitreolysis and pars plana vitrectomy in the treatment of vitreous floaters. METHODS: This is a single centre retrospective study of 31 patients (42 eyes) who underwent 54 procedures, Nd:YAG vitreolysis or pars plana vitrectomy, for the treatment of vitreous floaters between January 1992 and December 2000. Main outcome measures were percentage symptomatic improvement following treatment and incidence of post operative complications. Statistical analysis was performed using the Fisher exact test. RESULTS: Posterior vitreous detachment was the primary cause of floaters in all 42 eyes with co-existing vitreous veils in three eyes and asteroid hyalosis in two eyes. Thirty-nine of 42 eyes received Nd:YAG vitreolysis. Thirty-eight percent found Nd:YAG vitreolysis moderately improved their symptoms while 61.5% found no improvement. After an average of 14.7 months follow-up no post-operative complications were recorded. Fifteen eyes underwent a pars plana vitrectomy, one with combined phacoemulsification and posterior chamber implantation and 11 following unsuccessful laser vitreolysis. Pars plana vitrectomy resulted in full resolution of symptoms in 93.3% of eyes. One patient developed a post-operative retinal detachment which was successfully treated leaving the patient with 6/5 VA. CONCLUSION: Patients' symptoms from vitreous floaters are often underestimated resulting in no intervention. This paper shows Nd:YAG vitreolysis to be a safe but only moderately effective primary treatment conferring clinical benefit in one third of patients. Pars plana vitrectomy, while offering superior results, should be reserved for patients who remain markedly symptomatic following vitreolysis, until future studies further clarify its role in the treatment of patients with floaters and posterior vitreous detachment. PMID- 11913885 TI - Retinopathy of prematurity in the UK I: the organisation of services for screening and treatment. AB - AIMS: To ascertain how closely services for the screening and treatment of retinopathy of prematurity (ROP) were organised on a national level in 1995. METHODS: Questionnaires about the local arrangements for the screening and treatment of retinopathy of prematurity (ROP) were sent to the entire consultant membership (n = 648) of the Royal College of Ophthalmologists (RCOphth) and to the clinical directors (n = 259) of neonatal units and other units caring for preterm babies in the UK in 1995. RESULTS: 568/648 of UK consultants (88%) and 15 non-consultant ophthalmologists and 210/259 paediatricians (81%) and 19% paediatricians in non-neonatal units responded. Thirty-one per cent responding ophthalmologists were involved in the ROP service: of these 64% screened babies, 34% screened and treated babies, while 1% ophthalmologists treated ROP but did not screen. Ninety-six per cent units caring for preterm babies had their babies screened for ROP and for almost 95% of the screening took place in the neonatal unit. About 8200 babies were screened in 1994; 277 developed stage 3, of whom 54% received treatment. Nine per cent (n = 14) and 5% (n = 8) treated babies became blind in one and both eyes respectively. A sessional commitment was identified for 9% ophthalmologists, but for less than half this was included in the contracted work programme. Sixty-five ophthalmologists treated babies with ROP, but only 10 treated more than five babies in 1994. Training needs were identified by 71 respondents. CONCLUSIONS: Several aspects of ROP screening and treatment services require improvement. Hopefully, reducing the number of identified screeners would increase skills, confidence and the ability to recognise severe disease requiring treatment, and also facilitate incorporation of this work into consultant work plans. PMID- 11913887 TI - Quality of life in patients with early, moderate and advanced glaucoma. AB - PURPOSE: To evaluate the quality of life in glaucomatous patients using two different questionnaires: the medical outcomes study 36-item short-form health survey (MOS SF-36) and Viswanathan et al's questionnaire and to compare these two questionnaires. METHODS: Seventy-seven patients with glaucoma were consecutively selected. Two force-choice questionnaires were administered to each patient. Viswanathan et al's questionnaire was related to visual disability and the second was related to the quality of life from the MOS 36-item short-form health survey. Both questionnaires were evaluated among all the considered patients and the results were compared. Then the questionnaire which did the best evaluation was used to test the quality of life in three different subgroups based on the mean deviation of the worse eye. Mann-Whitney non parametric test and Spearman's r coefficient were used and a P value less than 0.05 was considered to be statistically significant. A linear regression model was used. RESULTS: In the entire group (n = 77) the Mean Deviation (MD) was -6.5 +/- 6.8 dB (mean +/- standard deviation) and Corrected Pattern Standard Deviation (CPSD) was 4.7 +/- 4.1 dB. The score of the Viswanathan et al's questionnaire was 8.3 +/- 2.4, while MOS SF-36 score ranged from 60.5% to 100% (mean score %). A significant (P < 0.0001) correlation was found between the score of the Viswanathan et al's questionnaire and MD (r = 0.79), Pattern Standard Deviation (PSD) (r = -0.68) and CPSD (r = -0.61). CONCLUSION: Viswanathan et al's questionnaire was more useful than MOS SF-36, both for the score and for the velocity to use. Furthermore Viswanathan et al's questionnaire was more significantly correlated to visual field MD. PMID- 11913886 TI - A short term study of the additive effect of timolol and brimonidine on intraocular pressure. AB - PURPOSE: To evaluate the additive ocular hypotensive effect of the combination of brimonidine and timolol on intraocular pressure (IOP) reduction in patients with glaucoma. METHODS: This was a prospective, randomized, double-masked, crossover study in 20 patients with primary open angle glaucoma (POAG) on therapy receiving timolol maleate 0.5% twice daily, with IOP greater than or equal to 22 mmHg in one eye. The treatment period was 3 weeks and during this period timolol + brimonidine or timolol + placebo were applied topically twice daily and IOP, blood pressure, heart rate and pupil size were measured. RESULTS: Combined therapy (timolol + brimonidine) had clinically significant IOP-lowering effect during the treatment period P << 0.01). The mean diurnal IOP was significantly reduced by an average of 5.1-5.9 mmHg (21.2-24.5%) compared with baseline value. The timolol + placebo combination had no clinically significant IOP-lowering effect (P > 0.05). No clinically significant side effects were observed during the treatment of both groups. CONCLUSIONS: This study showed that the combination of topically applied brimonidine and timolol cause a marked and sustained IOP reduction. PMID- 11913888 TI - Management of peri-ocular skin tumours by laissez-faire technique: analysis of functional and cosmetic results. AB - AIM: The role of wound healing by secondary intention in the treatment of peri ocular skin tumours is not well established. The object of this retrospective analysis was to evaluate the functional and cosmetic outcome of patients treated by the Laissez-faire technique in situations where primary closure would not have been possible. METHODS: Skin defects following excision of lid and peri-ocular tumours in 24 Caucasian patients were allowed to heal by granulation. The locations included lower eyelid (n = 10), upper lid (n = 6), medial canthus (n = 5), nasojugal fold (n = 2), lateral canthus (n = 1) and brow (n = 1). Four patients had lid margin involvement. The size of the initial defect, time taken to heal, discomfort during healing, the functional and cosmetic results-both from the surgeon and patient perspective, complications, secondary intervention if any and patient satisfaction were studied. RESULTS: A good functional and cosmetic result was obtained in 23 of the 25 lesions (92%). Of these 23 patients, two patients had slightly hypertrophied scars, which responded well to massage and two patients had some degree of ectropion. Of the two patients who did not have a good cosmetic result, only one needed secondary intervention. One had an exuberant granulation tissue, which responded to topical steroids and massage, but left behind a distorted lateral canthus. CONCLUSION: Healing by secondary intention of large defects following excision of peri-ocular tumours is an effective alternative to primary or staged reconstruction in selected cases. PMID- 11913889 TI - Characteristics and functional outcomes of 130 patients with keratoconus attending a specialist contact lens clinic. AB - AIMS: To evaluate the demographic profile, management and functional outcomes of patients with keratoconus attending the contact lens clinic of a tertiary ophthalmic referral centre over a one year period. METHODS: A retrospective cohort analysis was conducted by reviewing the computerised hospital records of 130 patients attending The Western Eye Hospital contact lens department over the period 1st Jan 1999 to 31st Dec 1999. Data on age, gender, referral pattern, visual acuity, contact lens fitting, degree of visual success, and some information on penetrating keratoplasty were obtained. RESULTS: 16.4% of all patients attending the Contact Lens clinic had keratoconus. The mean age at referral was 28.6 years and the mean age of keratoconus during the study period was 34.9 years. There was a predominance of male patients. Optometrists formed 72.2% of the referrals, and had prescribed some form of refractive correction in 70% of patients (two-thirds contact lenses) prior to hospital assessment. Of the 130 patients seen in the department during the study period, the post-referral management included bilateral contact lens fitting for 102 patients (78.5%), monocular contact lens fitting for 24 patients (18.5%) and no intervention in four patients (3%). The types of contact lenses used included PMMA lenses (2.7%), rigid gas permeable lenses (96.1%) of the spherical, elliptical and special cone lens designs, Keratosoft or Softperm lenses (0.8%) and scleral lenses (0.4%). Eleven eyes of eight patients had received penetrating keratoplasty (PK) prior to hospital assessment, of whom seven eyes needed post-surgical contact lens fitting. The main reasons for PK were contact lens intolerance (83%), frequent contact lens displacement (8.5%) and unsatisfactory visual acuity despite good contact lens fit (8.5%). Sixty-five per cent of patients were able to wear their contact lenses for more than 12 hours a day. With contact lens wear, 87% of patients had a visual acuity of 6/9 or better and 59% of eyes had improved visual acuity of 0.6 logMAR or more. CONCLUSION: Optometrists were the main source of referral for keratoconus patients to the Hospital Eye Service (HES). The mean age at referral was 28.6 years, with a predominance of male patients. Blurred vision formed the main presenting visual symptom on initial hospital assessment; subsequently, more than two-thirds of patients required bilateral contact lenses. Rigid gas permeable contact lenses remain the mainstay treatment for advanced keratoconus, with various designs enabling a large proportion of patients to attain improved visual acuity. PMID- 11913890 TI - Efficacy and safety of sedation with propofol in peribulbar anaesthesia. AB - PURPOSE: To evaluate the efficacy and safety of a sub-anaesthetic dose of propofol for reducing patient recall of peribulbar block in eye surgery. METHODS: A retrospective analysis of patients scheduled for elective cataract extraction or trabeculectomy using peribulbar anaesthesia with an intravenous bolus of propofol to provide sedation during the administration of the block. The dose of propofol was based on age and body weight. Patients' vital signs were monitored with continuous pulse oximetry and blood pressure measurements. Efficacy of sedation was assessed by recording patient's recall of the anaesthetic block after 8-10 min. RESULTS: Data from 2043 patients were analysed. The dose of propofol used ranged from 15-75 mg. Propofol was effective in abolishing recall in 87.5% of the patients studied. Only four patients required airway support but no major systemic side effects were encountered. CONCLUSIONS: A single sub anaesthetic dose of propofol prior to administering peribulbar block is effective in reducing recall of the injection and safe without major systemic side effects. PMID- 11913892 TI - Late surface opacification of Hydroview intraocular lenses. AB - PURPOSE: To describe clinical and pathological features of Hydroview intraocular lenses undergoing delayed surface opacification resulting in visual deterioration. METHODS: Twenty one eyes which underwent uncomplicated phacoemulsification and Hydroview lens implantation with good visual recovery, presenting at 46-146 weeks post-surgery with visual deterioration and glare symptoms resulting from opacification of the implants, were included in the study. Twelve eyes had severe opacification, of which nine underwent intraocular lens exchange and three more are still awaiting surgery. The method of explantation is described. The explanted intraocular lenses were examined using light microscopy, scanning electron microscopy and x-ray microanalysis using a light element detector. RESULTS: Light microscopy and scanning electron microscopy revealed diffuse granular deposits of approximately 5 microm diameter covering the optic surfaces but sparing the lens haptics. Light microscopic staining techniques and x-ray microanalysis confirm the major component of the deposits to be calcium phosphate salts. CONCLUSIONS: Late opacification of Hydroview intraocular lens implants is uncommon and aetiology seems to be multifactorial. Implant exchange is necessary to restore sight in some cases. As new materials are increasingly used it is important to highlight such unusual occurrences. PMID- 11913891 TI - Keratocyte apoptosis and corneal antioxidant enzyme activities after refractive corneal surgery. AB - PURPOSE: Refractive corneal surgery induces keratocyte apoptosis and generates reactive oxygen radicals (ROS) in the cornea. The purpose of the present study is to evaluate the correlation between keratocyte apoptosis and corneal antioxidant enzyme activities after different refractive surgical procedures in rabbits. METHODS: Rabbits were divided into six groups. All groups were compared with the control group (Group 1), after epithelial scraping (Group 2), epithelial scrape and photorefractive keratectomy (PRK) (traditional PRK: Group 3), transepithelial PRK (Group 4), creation of a corneal flap with microkeratome (Group 5) and laser assisted in situ keratomileusis (LASIK, Group 6). Terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labelling assay (to detect DNA fragmentation in situ) and light microscopy were used to detect apoptosis in rabbit eyes. Glutathione peroxidase (Gpx) and superoxide dismutase (SOD) activities of the corneal tissues were measured with spectrophotometric methods. RESULTS: Corneal Gpx and SOD activities decreased significantly in all groups when compared with the control group (P<0.05) and groups 2, 3 and 6 showed a significantly higher amount of keratocyte apoptosis (P<0.05). Not only a negative correlation was observed between corneal SOD activity and keratocyte apoptosis (cc: -0.3648) but Gpx activity also showed negative correlation with keratocyte apoptosis (cc: -0.3587). CONCLUSION: The present study illustrates the negative correlation between keratocyte apoptosis and corneal antioxidant enzyme activities. This finding suggests that ROS may be partly responsible for keratocyte apoptosis after refractive surgery. PMID- 11913893 TI - Symptomatic abnormalities of dark adaptation in patients with EFEMP1 retinal dystrophy (Malattia Leventinese/Doyne honeycomb retinal dystrophy). AB - PURPOSE: To investigate the nature of symptomatic visual disturbance in patients with EFEMP1 retinal dystrophy in the absence of geographic atrophy or choroidal neovascularization. METHODS: Patients presenting to a tertiary referral centre underwent clinical evaluation, fluorescein angiography, colour contrast sensitivity, focal, pattern, and standard electroretinography, electrooculography, scotopic threshold perimetry and dark adaptometry. RESULTS: Clinical features included reduced central vision, difficulty passing from light to dark, and diffuse submacular and peripapillary deposits, which were hyperfluorescent by fluorescein angiography. Colour contrast thresholds were abnormal in all six patients studied and both pattern and focal electroretinograms were abnormal in five of six patients. The scotopic and mixed rod-cone single flash ERG was normal but two patients demonstrated reduced oscillatory potentials and one had borderline delayed 30 Hz responses. Scotopic thresholds were elevated and rod-mediated dark adaptation kinetics were markedly prolonged in all six patients when measured over the central visible confluent deposits. CONCLUSIONS: In patients with EFEMP1 retinal dystrophy with confluent macular deposits, scotopic sensitivity is reduced and dark adaptation kinetics are prolonged over the macular deposits but are normal elsewhere. These results emphasize the localised nature of functional deficits in some patients with EFEMP1 retinal dystrophy and correlate well with the patient's visual symptoms. Symptomatic visual dysfunction may precede the development of clinically evident geographic atrophy or choroidal neovascularization in this disorder. PMID- 11913894 TI - Visual results in children treated for macular retinoblastoma. AB - PURPOSE: To evaluate the results of patching treatment in children with macular retinoblastoma in one eye. METHODS: Fifteen children affected by macular retinoblastoma received instructions for patching treatment for amblyopia. Data were collected on age at diagnosis of the tumor, presence of unilateral or bilateral disease, area of posterior pole involvement by the scar of the regressed tumor and its relationship to the fovea; and the onset, duration, and compliance of patching. The visual acuities recorded were expressed in logMAR (logarithm minimum angle of resolution) equivalents. RESULTS: Twelve children (80%) had bilateral retinoblastoma with the macular involved in one eye and three children had unilateral macular tumors. The median age at which patching was initiated was 15 months (range 4-36). Compliance to patching was good in 80% of children, with a median duration of 4 h (range 0.5-8) per day, 7 days per week, with total occlusion of the better eye. The median percentage of posterior pole involvement was 34% (range 11-100%). Eighty percent of children had some improvement in their visual acuity, and of the children in whom final logMAR acuity was recorded, 73% had an acuity of 1.0 logMAR or better and 53% an acuity of 0.5 logMAR or better after patching. There was no evidence of association between age of patient, sex, duration of patching, or percentage of posterior pole involvement and the improvement in visual acuity. CONCLUSIONS: In spite of the macular involvement of eyes with retinoblastoma, some visual recovery was achieved in 80% of children. Hence a trial of patching therapy is recommended for all children with involvement of the macula by retinoblastoma. PMID- 11913895 TI - Inflammatory pigmented paravenous retinochoroidal atrophy. AB - PURPOSE: To describe an active inflammatory cause of pigmented paravenous retinochoroidal atrophy. METHODS: A 54-year-old female patient presented with complaints of worsening visual acuity and poor night vision was examined. Fundus examination was performed and color fundus photographs were taken. In addition to fluorescein angiography, visual field examinations and electroretinographic tests were performed. Macular evaluation was performed with optical coherence tomography. RESULTS: Both fundi showed circumscribed patches of retinochoroidal atrophy and pigmentation along the retinal veins. She had also marked vitreous cells with snow ball opacities and cystoid macular edema in both eyes. Fluorescein angiography confirmed the presence of a hyperfluorescence due to widespread paravenous retinal pigment epithelial defect while ICG angiography disclosed hypofluorescence in all phases. The electroretinogram showed reduced responses especially in the left eye. Visual field tests showed scotomas corresponding with areas of atrophy along the retinal veins. CONCLUSIONS: This is a report of the findings in pigmented paravenous retinochoroidal atrophy that is a nonspecific degenerative disease and may occur in association with systemic infections or inflammation. Ocular inflammation with cystoid macular edema is an unusual manifestation of the disease. PMID- 11913896 TI - Migration of seton into the anterior chamber. PMID- 11913898 TI - Nasolacrimal duct obstruction following chicken pox. PMID- 11913897 TI - Progressive ophthalmoplegia in arthrogryposis multiplex congentia. PMID- 11913900 TI - Choroidal telangiectasia in a patient with hereditary hemorrhagic telangiectasia. PMID- 11913899 TI - Biofilm formation and coccal organisms in infectious crystalline keratopathy. PMID- 11913901 TI - Retained fragments in the anterior segment following phacoemulsification surgery. PMID- 11913902 TI - Plasmin-assisted vitrectomy eliminates cortical vitreous remnants. PMID- 11913903 TI - Central retinal vein occlusion and thrombophilia. AB - Central retinal vein occlusion is one of the commonest vascular diseases of the eye. The pathogenesis is multifactorial with both local factors and systemic diseases being aetiologically important. Many thrombophilic conditions have recently been identified and studies looking at their potential role in CRVO have been undertaken. The aim of this review is to critically appraise these studies as to date many have given conflicting results, making it far from clear what role thrombophilic conditions play in CRVO. It appears that hyperhomocysteinaemia and antiphospholipid syndrome are causes of CRVO and there is evidence that disorders causing hypofibrinolysis may also be important. The common hereditary thrombophilic conditions however do not appear to be strong risk factors but larger studies are needed for a definitive answer. PMID- 11913905 TI - Drug problem recognition, desire for help, and treatment readiness in a soup kitchen population. AB - This study determined hypothesized predictors of three components of motivation for change--drug problem recognition, desire for help, and treatment readiness- in a high-risk, drug-using population. The sample consisted of 190 guests at two inner-city soup kitchens in Brooklyn, NY who reported drug/alcohol use and were not participating in substance dependency treatment. Ever receiving addiction treatment, having no trade/job skills, and more severe symptoms of depression were associated with greater drug problem recognition. More recent days of drug/alcohol use, intensive pattern of drug use, and greater problem recognition were associated with greater desire for help. Caring for children, more recent days of drug/alcohol use, physical health problems, and desire for help were the direct predictors of treatment readiness. Problem recognition had a strong indirect effect on readiness mediated through desire for help. Knowledge of a drug user's motivational state and the factors leading to it can help guide the development of more effective interventions. PMID- 11913904 TI - Revisiting sub-Saharan African countries' drug problems: health, social, economic costs, and drug control policy. AB - This article takes an international perspective on the drug problem in sub Saharan Africa. This analysis borrows ideas from physical and economic geography as a heuristic device to conceptualize the global narcoscapes in which drug trafficking occurs. Both the legitimate and the illegal drug trade operate within the same global capitalist system and draw on the same technological innovations and business processes. Central to the paper's argument is evidence that sub Saharan African countries are now integrated into the political economy of drug consumption due to the spill-over effect. These countries are now minor markets for "hard drugs" as the result of the activities of organizations and individual traffickers that use Africa as a staging point in their trade with Europe and the United States. As a result, sub-Saharan African countries have drug consumption problems that were essentially absent prior to 1980, along with associated health, social, and economic costs. The emerging drug problem has forced African countries to develop their own drug control policy. The sub-Saharan African countries mentioned below vary to some extent in the level of drug use and misuse problems: Burundi, Comoros, Djibouti, Eritrea, Ethiopia, Kenya, Madagascar, Malawi, Mauritius, Mozambique, Reunion, Rwanda, Seychelles, Somalia, Tanzania, Uganda, Zambia, Angola, Cameroon, Central African Republic, Chad, Congo, Congo (Zaire), Equatorial Guinea, Gabon, Sao Tome and Principe, Botswana, Lesotho, Namibia, South Africa, Swaziland, Benin, Burkina Faso, Cape Verde, Cote d'Ivoire, Gambia, Ghana, Guinea, Guinea Bissau, Liberia, Mali, Mauritania, Niger, Nigeria, Senegal, Sierra Leone, and Togo. As part of this effort, African countries are assessing the health, social, and economic costs of drug-use-related problems to pinpoint methods which are both effective and inexpensive, since their budgets for social programs are severely constrained. Many have progressed to the point of adopting anti-drug laws or legislation, or of establishing a drug control agency. They are also cooperating regionally to coordinate drug control measures and working with the Organization of African Unity (OAU). In addition, almost all the sub-Saharan African countries are signatories to all United Nations drug conventions. Since the drug problem in Africa has international origins, it will take concerted international cooperation and coordinated effort to combat the "social cancer" of drugs. PMID- 11913906 TI - Determinants of youth smoking--evidence from Turkey. AB - This paper investigates the determinants of smoking among middle- and high-school students in Ankara, Turkey. Data were collected from 4800 students in the second semester of the 1997-98 academic year to investigate substance use prevalence among young people. In addition to classical independent variables, factors which are not frequently used in the literature--such as school type, neighborhood, family religiosity, and factors reflecting the cultural background of the students--are included in the analysis. The results of binomial logistic regression offer evidence for the effects of school type, a smoking-related attitude, presence of a stepmother, father's use of alcohol, sister and brother who smoke, student's alcohol use, and participation in art activities. PMID- 11913907 TI - Drug use and suicide attempts: the role of personality factors. AB - This study on suicide attempts is part of a large research project on dependent behavior in adolescents and young adults. 228 subjects aged 14-25 (107 "drug abusers," 121 controls) from the French speaking part of Switzerland were evaluated on the basis of a semi-structured interview (Mini International Neuropsychiatric Interview), enabling a DSM-IV diagnosis, and self-reports (SSS by Zuckermann, MMPI-2, IDI by Hirschfeld). 31.5% of "drug abuser" males and 41.2% of "drug abuser" females committed one or more suicide attempts. The results of a logistic regression show that the significant factor explaining suicide attempts in drug users is emotional reliance for males and experience-seeking for females. PMID- 11913908 TI - The validity of a social indicator approach to substance misuse needs assessment. AB - Factors other than the magnitude of a community's drug- or alcohol-use-related problems may play a large role in resource allocation. Needs assessment approaches such as key informant interviews and household surveys can contribute to a more informed process. An additional approach to needs assessment, social indicator modeling, addresses the high cost of surveys and the potential subjectivity of key informant interviews. This paper discusses the history of social indicator approaches to needs assessment and reviews recent equitable distribution approaches. An equitable distribution model of alcohol- and other drug-use-related problems was created and validated. The drug model accounted for close to 50% of the variation in estimates of lifetime diagnosis of drug dependence or misuse. The alcohol model performed less successfully, accounting for 21% of the variation in lifetime diagnosis of alcohol-use-related problems. PMID- 11913909 TI - Use of psychoactive substances and sexual risk behavior in adolescents. AB - This study investigated the relationship between the use of illicit drugs and sexual-risk-behavior in a sample of students aged 14 to 21 years at a public high school in Sao Paulo in 1997. A total of 689 useable questionnaires documented the sample in consumption of psychoactive substances and sexual behavior. Sexual behavior of drug users and non-users was compared regarding history of complete sexual intercourse, age at first sexual intercourse, use of condoms, sexual intercourse with sex workers, and prostitution. Drug users (N = 366) presented a higher frequency of complete sexual intercourse (80.8% of users versus 53.5% of non-users), (N = 323, p < .001), a younger age at first sexual intercourse (on average 15.2 years in users versus 15.7 among non-users, p < .005), a trend toward lower use of condoms (56.7% among users versus 65.3% among non-users, p < .1), and more sexual intercourse with sex workers (31.1% among users versus 15% among non-users, p < .001). Sexual-risk-behavior increased with the number of drugs used. Alcohol and marijuana use were associated with the highest sexual risk-behavior. These data are essential for the development of more specific preventive strategies, focusing on male alcohol and marijuana users. PMID- 11913910 TI - Colorectal cancer prevention. PMID- 11913911 TI - Sedation for endoscopy. PMID- 11913912 TI - Sudden cardiac death in the young. PMID- 11913913 TI - Colonoscopic surveillance for family history of colorectal cancer: are NHMRC guidelines being followed? AB - OBJECTIVES: To assess whether referrals for surveillance colonoscopy and subsequent follow-up recommendations for patients with a family history of colorectal cancer concurred with the published National Health and Medical Research Council (NHMRC) guidelines. DESIGN: A prospective audit of patients with a family history of colorectal cancer referred for surveillance colonoscopy. Follow-up recommendations were assessed retrospectively. SETTING AND SUBJECTS: All patients referred to a major teaching hospital for surveillance colonoscopy on the basis of a family history of colorectal cancer from 2 January 2000-15 April 2001. MAIN OUTCOME MEASURES: Concurrence of referrals and recommendations with NHMRC guidelines. RESULTS: Of 340 patients referred because of a family history of colorectal cancer, 202 (83 men, 119 women) were asymptomatic. Their mean age was 50 years (95% CI, 48.3-51.6 years). The family history of 95 (47%) of these patients satisfied the NHMRC criteria for colonoscopic surveillance. Another 20 patients (17%) satisfied the criteria, but were referred before the recommended age to commence surveillance. Analysis by referral source showed that the proportion of referrals meeting NHMRC guidelines was higher from specialists than from general practitioners (75% v 45%), and this difference was significant. Follow-up recommendations, when made, concurred with NHMRC guidelines in 81% of cases. CONCLUSIONS: Further education of the medical community is required to increase understanding of colorectal screening strategies and ensure appropriate resource allocation. PMID- 11913914 TI - Applying evidence-based guidelines improves use of colonoscopy resources in patients with a moderate risk of colorectal neoplasia. AB - OBJECTIVES: To determine whether applying National Health and Medical Research Council (NHMRC) guidelines for colorectal cancer prevention would reduce the number of follow-up colonoscopies. DESIGN: A prospective audit of colonoscopic surveillance decisions before and after the intervention. SETTING: The endoscopy suite at a metropolitan tertiary hospital three months before and after January 2000. INTERVENTION: Dissemination of NHMRC guidelines, and supervision of application of the guidelines by a nurse coordinator. SUBJECTS: We compared colonoscopic surveillance decisions before and after the intervention in two groups of 100 consecutive patients after polypectomy and in two groups of 50 consecutive patients with a family history of colorectal cancer after a normal colonoscopy. MAIN OUTCOME MEASURES: Change in concordance of decisions with NHMRC guidelines; and effect on number of follow-up colonoscopies. RESULTS: After the intervention, the proportion of postpolypectomy surveillance decisions matching the guidelines increased from 37% to 96% (P < 0.05). The mean time to repeat colonoscopy after polypectomy increased from 2.7 to 3.5 years (P < 0.005) (ie, a 23% reduction in the number of postpolypectomy surveillance colonoscopies performed per year). Likewise, the proportion of family-history surveillance decisions matching the guidelines increased from 63% to 96%. Adhering to the guidelines resulted in a 17% reduction in colonoscopies performed on the basis of a family history of colorectal cancer. CONCLUSIONS: Supervised application of evidence-based guidelines to a colorectal cancer surveillance program significantly reduces the number of surveillance colonoscopies performed. PMID- 11913915 TI - Sedation for endoscopy: the safe use of propofol by general practitioner sedationists. AB - OBJECTIVE: To determine the incidence of adverse events related to an endoscopy sedation regimen that included propofol, delivered by general practitioner (GP) sedationists. DESIGN: Audit of reports of sedation-related adverse events in patients undergoing endoscopy. A sample of 1000 patients' medical records was also reviewed to determine the drugs and dosages used and the proportion of sedations delivered by GPs. SETTING AND PARTICIPANTS: All patients undergoing gastroscopy and/or colonoscopy from January 1996 to December 2000 in two private endoscopy centres in Canberra. Sedation was provided by GPs or a specialist anaesthetist, in most cases using a drug regimen that included propofol. MAIN OUTCOME MEASURES: Incidences of respiratory arrest, airway obstruction, hypoxia requiring intervention, hypotension, and death; number of interventions to correct these events, including extra airway management, bag-mask ventilation, intravenous fluid infusion, endotracheal intubation and the use of reversal agents, and admission to hospital. RESULTS: 28,472 procedures were performed in the five years. There were 185 sedation-related adverse events (6.5/1000 procedures; 95% CI, 5.6-7.4): 107 for airway or ventilation problems (3.8/1000) and 77 hypotensive episodes (2.7/1000). Respiratory-related adverse events were more common in patients managed by GPs than anaesthetists, but this was not significant (P = 0.1). Interventions were recorded in 234 patients (8.2/1000; 95% CI, 7.2-9.3): 123 to maintain ventilation, and 111 intravenous infusions. GPs were more likely than anaesthetists to intervene to manage respiratory-related adverse events (P = 0.03). Four patients required transfer or admission to hospital. No patients required endotracheal intubation, and there were no deaths. CONCLUSIONS: The GP sedationists encountered a low incidence of adverse events, which they managed effectively. It appears that appropriately selected and trained GPs can safely use propofol for sedation during endoscopy. PMID- 11913916 TI - Paracetamol recall: a natural experiment influencing analgesic poisoning. AB - OBJECTIVES: To determine whether the occurrence of paracetamol and non paracetamol analgesic deliberate self-poisoning (DSP) and accidental paediatric poisoning was affected by two periods of recall of paracetamol products. DESIGN: Retrospective, observational audit of proportions of poisonings with tablet and capsule formulations of paracetamol, ibuprofen and aspirin products during two recall periods compared with the number of poisonings during the same periods of the previous three years. SETTING: A national poisons information centre and a regional toxicology service. MAIN OUTCOME MEASURES: Rates of DSP and accidental paediatric poisoning with paracetamol, ibuprofen and aspirin. RESULTS: During the two recall periods, there was a significant increase in ibuprofen DSP calls to the poisons information centre (RR, 1.86; 95% Cl, 1.41-2.44; P = 0.001). There was no significant change in paracetamol or aspirin DSP calls over the two recall periods. However, there was a non-significant reduction in DSP calls with paracetamol in the first recall period alone (P = 0.057). There was a significant increase in the proportion of aspirin DSP presentations for the toxicology service (RR, 3.33; 95% CI, 0.97-11.4; P = 0.043), but no significant changes in paracetamol and ibuprofen DSP presentations. For accidental paediatric ingestions there was a significant increase in the proportion of ibuprofen calls (RR, 2.35; 95% CI, 1.85-2.98; P = 0.001), but no significant change in paracetamol or aspirin calls. CONCLUSIONS: Reduced paracetamol availability increased poisoning with alternative analgesics, but had little effect on the incidence of paracetamol poisoning. Restriction of paracetamol-containing products may inadvertently increase poisoning with potentially more toxic agents. PMID- 11913917 TI - Severe opiate withdrawal in a heroin user precipitated by a massive buprenorphine dose. AB - By diverting his dispensed medication, our patient collected 11 buprenorphine tablets (8 mg each), which he took in one day. The result was not respiratory depression, but instead severe opiate withdrawal lasting four days--this scenario has not previously been reported. This case highlights features of the unique pharmacology of buprenorphine and some key issues for its use in the management of heroin dependence. PMID- 11913918 TI - The role of corticosteroids in the management of childhood asthma. AB - Preventive treatment: * Inhaled corticosteroids are indicated in children with asthma who have more than mild persistent asthma or are unresponsive to non steroidal medications after 2-4 weeks. * Initial administration of 400 microg/day of chlorofluorocarbon-beclomethasone dipropionate, or budesonide, or 200 microg/day of fluticasone propionate or hydrofluoroalkane-beclomethasone dipropionate, is suggested, with subsequent titration of the dose to achieve ongoing control with the lowest dose possible. * In situations where asthma control cannot be achieved with the above doses of inhaled corticosteroids, the addition of a long-acting beta2-agonist, theophylline or a leukotriene antagonist should be considered. * Specialist referral is recommended in children requiring high doses of inhaled steroids, regular oral steroids or in whom there is concern about possible steroid side effects. Treatment of acute asthma: * Systemic corticosteroid therapy is recommended for children with moderate to severe acute asthma or if there is incomplete response to beta2-agonists. * Initial administration of 1 mg/kg prednisolone (maximum, 50 mg) orally is suggested, and this may be repeated every 12-24 hours, depending on response. While a course of up to three days is generally sufficient, in more severe cases a prolonged course (with tapering) may occasionally be indicated. * The need for recurrent systemic corticosteroid therapy for acute episodes is an indication for reassessment of the child's interval therapy. PMID- 11913919 TI - The challenge of cultural and ethical pluralism to medical practice. AB - * "Culture" can be understood as the way in which people make sense of the world by deploying shared meanings, attitudes, assumptions and values. * Doctors will frequently encounter patients whose lives are guided by ethical systems and values that are different from their own. * Individuals may differ in their beliefs about decision-making, regardless of their cultural background. * Doctors should be willing to examine and test their own moral systems and cultural assumptions and be open to alternative traditions and beliefs. * Engaging with other cultures does not imply that all cultural norms should be accepted uncritically, as there may not always be room for compromise. * Failure to engage with issues of culture can erode the trust on which the doctor-patient relationship depends. * Tensions can only be resolved through rigorous attention to a person's story. PMID- 11913920 TI - Howard Florey, Alexander Fleming and the fairy tale of penicillin. AB - The public myth of the discovery of penicillin is an archetypal "quest story" of the type common to every human culture. But the real story of the discovery, testing and refinement of penicillin is a complex tale of accident, serendipity, oversight, conflict, the pressure of war, idiosyncratic personalities and even- the invention of history. PMID- 11913921 TI - Natural remedies for osteoporosis in postmenopausal women. PMID- 11913922 TI - Evidence of human metapneumovirus in Australian children. PMID- 11913923 TI - Risk of death from methicillin-resistant Staphylococcus aureus bacteraemia: a meta-analysis. PMID- 11913924 TI - PBS/RPBS cost implications of trends and guideline recommendations in the pharmacological management of hypertension. PMID- 11913925 TI - MEDicine or MADness. PMID- 11913926 TI - HIV among injecting drug users of Indo-Chinese ethnicity in Victoria. PMID- 11913927 TI - Detecting and reducing hospital adverse events: outcomes of the Wimmera clinical risk management program. PMID- 11913928 TI - SPHERE: a national depression project. PMID- 11913929 TI - EBM in action: is laser treatment effective and safe for musculoskeletal pain? PMID- 11913931 TI - Books as carriers of disease. PMID- 11913932 TI - Words alone are not enough. PMID- 11913933 TI - PHARMACopsychiatry: problematic but promising. PMID- 11913934 TI - Gastro-oesophageal malignancy in New Zealand: 1995-97. AB - AIM: To assess the incidence, treatment and survival of patients with oesophago gastric carcinoma in New Zealand. METHODS: All cases of oesophageal or gastric carcinoma diagnosed in 1995-97 were retrieved from the national cancer registry. Linked data describing all episodes of inpatient treatment for these patients were obtained from the New Zealand Health Information System. An analysis of demographics, treatment and survival was performed. RESULTS: A total of 1791 cases were recorded (616 oesophageal, 1175 gastric). Carcinomas of the gastrooesophageal junction made up the majority of cases. 1138 were male. The median age was 71 years. 78.6% were of European descent, 10.4% Maori, 3.6% Pacific Islanders and 7.4% of other ethnic backgrounds. There were a total of 3403 hospital admissions (median 1.0 per patient). Overall, 29% underwent a surgical resection (15% oesophageal, 36% gastric). Exploratory surgery alone was performed in 14% operated on for oesophageal cancer and 12.3% for gastric neoplasms. Following resection 90 day mortality was 11.8% (10.5% oesophageal, 12% gastric). Overall median survival was 6.3 months (5.8 months oesophageal, 6.6 months gastric) with 16.7% of patients alive at three years. Following resection median survival was 17.8 months ( 16.2 months oesophageal, 18.1 months gastric) with 35.8% of patients alive at three years (34.7% oesophageal, 36% gastric). CONCLUSIONS: These data provide a baseline for future studies of the evaluation and treatment of gastrooesophageal malignancy in New Zealand. PMID- 11913935 TI - Management of acute otitis media by New Zealand general practitioners. AB - AIMS: To determine the current management of acute otitis media by New Zealand general practitioners (GPs). METHODS: A reply-paid questionnaire was sent to 2000 New Zealand GPs. The questions relate to management of a three year old child presenting with her first episode of acute otitis media. RESULTS: 95% of respondents reported they would usually or always use antibiotics. Amoxicillin was the antibiotic of choice, followed by amoxicillin/clavulanate. Cotrimoxazole was the antibiotic of choice in the case of allergy to amoxicillin. 82% of respondents recommended follow-up, with a broad range of follow-up times (24 hours to 12 weeks). Approximately half of practitioners considered 5-6 episodes of acute otitis media in a year as an appropriate threshold for referral for grommets. Most GPs had received an update on otitis media within the previous two years. CONCLUSIONS: There is considerable variation in the management of acute otitis media by New Zealand GPs. Use of a national guideline may result in a more standardised, rational approach to the treatment of acute otitis media. PMID- 11913936 TI - Postoperative epidural analgesia following elective major abdominal surgery in high risk patients: a retrospective cohort study. AB - AIM: To describe the effect of post-operative epidural analgesia on morbidity and mortality rates in a group of high-risk patients undergoing elective major abdominal surgery. METHODS: Retrospective chart review of patients in American Society of Anaesthetists Physical Status (ASA) category III or IV, who underwent elective major I or II general surgical procedures between 01/01/1996 and 01/09/1998. Patients were identified from a prospective audit database. Patients who had epidural analgesia or conventional parenteral opioids were compared for outcome measures. RESULTS: There were 167 patients identified (72 epidural, 95 non-epidural group). There was no significant difference in demographic data, inpatient stay, intensive care unit stay, or mortality rates (11% epidural v 17% non-epidural, p>0.05). There was no significant difference in morbidity rates, however there was a non-significant trend towards a lower morbidity in the epidural group. CONCLUSIONS: This study does not show any benefit from post operative epidural analgesia on morbidity and mortality rates in high risk patients undergoing major abdominal surgery. It does illustrate that ASA 3 and 4 patients undergoing major abdominal surgery have a high morbidity and mortality. PMID- 11913937 TI - Drinking patterns among older people in the community: hidden from medical attention? AB - AIMS: To determine patterns of alcohol use and misuse among community-dwelling people aged 65 years and over in Christchurch and to assess how often this comes to medical attention. METHODS: A cross-sectional survey of alcohol use and misuse was conducted followed by a self-administered postal survey among non- respondents. General practitioners (GPs) of the respondents completed a self administered questionnaire on patients' alcohol use and misuse. RESULTS: The response rate was 58% (141/243). The prevalence of hazardous alcohol consumption in the past twelve months (AUDIT cut-off score 8 or more) was 9.9% (95% CI = 4.9 14.9) and the prevalence of lifetime alcohol dependence using DSM-IV diagnostic criteria was 24.8% (95% CI = 17.6-32.0). Men were more likely than women to report lifetime dependency and current hazardous patterns. The response rate among GPs was 77.7% (108/139). None of the GPs identified or diagnosed alcohol problems in the past twelve months among this group and reported a history of alcohol problems in only four (4.0%) patients. Those with current hazardous patterns of alcohol use were twice as likely to be admitted to hospital (RR=2.4; 95% CI 1.2-5.1) but significantly less likely to visit their GPs in the previous twelve months (RR=0.55; 95% CI 0.7-1.1). CONCLUSION: A significant proportion of community-dwelling elderly people reported patterns of alcohol consumption that put them at risk of future damage to physical or mental health. Hazardous drinkers were less likely to visit their GPs and only in a few cases, were GPs aware of such potential problems. PMID- 11913938 TI - The role of the cytokine milieu in protecting us from allergy. PMID- 11913939 TI - Inadequate availability of pharmacological treatment for affective disorders in New Zealand. AB - Both BAD and unipolar depression are common, serious and, in some cases, chronic conditions which often require long term pharmacotherapy. Drug side effects can be severe, contribute to poor compliance and are therefore a factor in maintaining illness and precipitating relapse. Treatment resistance is common. Current evidence suggests that a number of pharmacological agents which are available in other countries (and in some cases have been for many years) may be useful when side effects or treatment resistance occur. In some cases these alternatives are part of recommended algorithms for treatment. In New Zealand, when side effects occur, the options are limited and the problem of treatment resistance is usually approached by using poorly researched combinations of the currently available drugs, thus increasing the likelihood of side effects and adverse drug interactions. The current system is failing to provide appropriate or adequate treatment for people suffering from affective disorders and should be urgently reviewed. PMID- 11913940 TI - Reducing the uncertainties of withdrawing or withholding treatment. PMID- 11913941 TI - What's happening in New Zealand emergency departments? PMID- 11913942 TI - Liability of health professionals for a breach of the abortion law in New Zealand. PMID- 11913943 TI - Pseudoephedrine causing mania-like symptoms. PMID- 11913944 TI - B cell signaling. Introduction. AB - The B cell antigen receptor (BCR) is composed of the membrane form of the immunoglobulin (Ig) and the Ig-alpha/Ig-beta heterodimer, which function as the antigen recognition component and the signaling component, respectively. A signal transmitted by BCR modulates gene expression, adhesion or survival, thereby determining the fate of antigen-encountered B cells. BCR proximal signaling occurs within cholesterol- and sphingolipid-rich plasma membrane microdomains termed lipid rafts, and involves tyrosine kinases such as Lyn, Syk and Btk and the adapter molecule SLP65/BLNK. Although the distal signaling cascades via BCR are not yet fully elucidated, various components are already identified, such as lipid kinases and small G-proteins. BCR signaling is regulated by various membrane molecules termed co-receptors such as CD19 and CD22. The BCR co receptors appear to be required for normal immune functions. Viral proteins such as LMP2 also regulate BCR signaling to maintain viral latency. Various aspects of BCR signaling and its regulatory mechanisms are discussed in this issue. PMID- 11913945 TI - Multitasking of Ig-alpha and Ig-beta to regulate B cell antigen receptor function. AB - Since their discovery as signaling subunits of the B cell antigen receptor (BCR), Ig-alpha and Ig-beta are discussed to serve either a redundant or distinct function for B cell development, maintenance, and activation. Dependent upon the experimental system that has been used to address this issue, evidence could be provided to support both possibilities. Only recently has it become clear that Ig alpha and Ig-beta possess a unique signaling identity but that both together are required to orchestrate proper B cell function in vivo. Here we discuss some of the underlying mechanisms that may involve direct coupling to discrete subsets of BCR effector proteins, such as protein tyrosine kinases or the intracellular adaptor SLP-65/BLNK. PMID- 11913946 TI - Regulation of phospholipase C-gamma2 and phosphoinositide 3-kinase pathways by adaptor proteins in B lymphocytes. AB - The importance of phosphoinositide 3-kinase (PI3K) and phospholipase C (PLC) gamma2 in B cell function and development has been highlighted by gene targeting experiments in mice. In fact, these knockout mice exhibit a profound inhibition of proliferative responses upon B cell receptor (BCR) engagement. The molecular connections between these effectors and upstream tyrosine kinases such as Syk have been studied intensively in the past few years. This mechanism involves the action of cytoplasmic adaptor molecules, which participate in forming multicomponent signaling complexes, thereby directing the appropriate subcellular localization of effector enzymes. In addition to these cytoplasmic adaptor proteins, cell surface coreceptors can be viewed as transmembrane adaptor proteins, because coreceptors can also change the localization of effector enzymes, which in turn modulates the BCR-initiated signals. PMID- 11913947 TI - The role of the protein tyrosine phosphatase CD45 in regulation of B lymphocyte activation. AB - The transmembrane protein tyrosine phosphatase CD45 is expressed throughout B cell development and differentiation, with the exception of terminally differentiated plasma cells on which its expression is down regulated. Numerous studies using CD45-deficient B cell lines and CD45-deficient mice have clearly demonstrated that CD45 plays an important role in modulating the signal that is transduced via the B cell antigen receptor by regulating the phosphorylation state of Src family kinases. Spatial and temporal controls enable CD45 to promote B cell antigen receptor signal transduction by constitutively maintaining Src family kinases in a partially active state, such that the B cell is able to effectively respond to an antigenic challenge. Moreover, CD45 is required for optimal activation of Ca2+-dependent and MAP kinase-dependent signal transduction pathways in the B cell. The net result is that CD45 affects the B cell response by controlling the relative threshold of sensitivity to a given antigenic stimulus. Thus, CD45 expression and function is required for normal B cell development, tolerance induction, and responsiveness to antigen. PMID- 11913948 TI - CD19, CD21, and CD22: multifaceted response regulators of B lymphocyte signal transduction. AB - B lymphocyte development and function depend upon the activity of intrinsic and B cell antigen receptor (BCR)-induced signals. These signals are interpreted, amplified, fine-tuned, or suppressed through the precise actions of specialized cell surface coreceptors, or "response regulators," that inform B cells of their extracellular environment. Important cell surface response regulators include the CD19/CD21 complex, CD22, and CD72. CD19 establishes a novel Src-family protein tyrosine kinase (PTK) amplification loop that regulates basal signaling thresholds and intensifies Src-family PTK activation following BCR ligation. In turn, CD22 limits the intensity of CD19-dependent, BCR-generated signals through the recruitment of potent phosphotyrosine and phosphoinositide phosphatases. Herein we discuss our current understanding of how CD19/CD21 and CD22 govern the emergence and intensity of BCR-mediated signals, and how alterations in these tightly controlled regulatory activities contribute to autoimmunity in mice and humans. PMID- 11913949 TI - Activation and function of the Rap1 GTPase in B lymphocytes. AB - Rap1 is a monomeric GTPase that is closely related to Ras. In this review, we summarize our recent work showing that the B cell antigen receptor (BCR), as well as chemokine receptors, activate Rap1 via a pathway that involves phospholipase C dependent production of diacylglycerol (DAG). The possible identities of the DAG regulated guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) that regulate the activation of Rap1 by the BCR and chemokine receptors will be discussed. Although initially thought to be an antagonist of Ras-mediated signaling, Rap1 does not appear to modulate the ability of the BCR to activate downstream targets of Ras. Instead, activation of Rap1 promotes B cell adhesion as well as B cell migration toward chemokines. Thus, Rap1 may play a key role in a number of processes that are essential for B cell development and activation. PMID- 11913950 TI - Molecular mechanisms for apoptosis induced by signaling through the B cell antigen receptor. AB - Although the B cell antigen receptor (BCR) transmits survival and activation signals, BCR ligation can induce apoptosis in both immature and mature B cells. BCR-mediated apoptosis is suggested to play a role in self-tolerance by deleting self-reactive B cells. Generation of an apoptotic signal through BCR appears to depend on the composition of the higher order BCR complex and is suggested to occur outside the plasma membrane microdomains, termed lipid rafts. During BCR mediated apoptosis, mitochondrial dysfunction is induced and is essential for apoptosis, probably by activating both caspases, cysteine proteases that play a central role in apoptosis, and caspase-independent effectors for apoptosis. Although signaling pathways for apoptosis are not yet fully defined in BCR mediated apoptosis, expression of the proto-oncogene product c-Myc is enhanced upon BCR ligation, and c-Myc appears to mediate BCR ligation-induced apoptosis by causing mitochondrial dysfunction, suggesting that BCR-mediated apoptosis is a form of Myc-induced apoptosis. PMID- 11913951 TI - The effects of the Epstein-Barr virus latent membrane protein 2A on B cell function. AB - Epstein-Barr Virus (EBV) infects B-lymphocytes circulating through the oral epithelium and establishes a lifelong latent infection in a subset of mature memory B cells. In these latently infected B cells, EBV exhibits limited gene expression with the latent membrane protein 2A (LMP2A) being the most consistently detected transcript. This persistent expression, coupled with many studies ofthe function of LMP2A in vitro and invivo, indicates that LMP2A is functioning to control some aspect of viral latency. Establishment and maintenance of viral latency requires exquisite manipulation of normal B cell signaling and function. LMP2A is capable of blocking normal B cell signal transduction in vitro, suggesting that LMP2A may act to regulate lytic activation from latency in vivo. Furthermore, LMP2A is capable of providing B cells with survival signals in the absence of normal BCR signaling. These data show that LMP2A may help EBV-infected cells to persist in vivo. This review discusses the advances that have been made in our understanding of LMP2A and the effects it has on B cell development, activation, and viral latency. PMID- 11913952 TI - Conserved cellular and molecular mechanisms in development. AB - This review discusses examples of conserved cellular and molecular mechansims in development, including the pathway of signal transduction between the photoreceptors R8 and R7 in Drosophila, which is compared to vulval induction in Caenorhabditis elegans. The Wg pathway in Drosophila is compared, first, to the Wnt pathway in dorsal mesoderm specification in Xenopus: second, to the same pathway in sea urchins; third, to the equivalent in the mom cascade of C. elegans; and finally, to parts of the equivalent pathway in Dictyostelium discoideum. The conserved expression of some hox genes in vertebrate limb buds and in the heads or tails of several invertebrate and vertebrate embryos is also illustrated. Two further examples show the contrast between BMP4 and the proteins Noggin, Chordin, Follistatin and Gremlin. Another example concerns the binding of proteins to the 3'UTRs of several messages which inhibits their translation in Drosophila and C. elegans. The final example illustrates the selective transfer of transcription factors to nuclei in Drosophila and C. elegans. PMID- 11913953 TI - Ultrasensitivity in (supra)molecularly organized and crowded environments. AB - The ultrasensitive response of biological systems is a more sensitive one than that expected from the classical hyperbola of Michaelis-Menten kinetics, and whose physiological relevance depends upon the range of variation of substrate or effector for which ultrasensitivity is observed. Triggering and modulation of the ultrasensitive response in enzymatic and cellular systems are reviewed. Several demonstrations of ultrasensitive behavior in cellular systems and its impact on the amplification properties in signalling cascades and metabolic pathways are also highlighted. It is shown that ubiquitous cytoskeletal proteins may up- or downmodulate ultrasensitivity under physico-chemical conditions resembling those predominant in cells. PMID- 11913954 TI - Analysis of p53-dependent mechanisms in the maintenance of genetic stability in diploid tumourigenic line SK-UT-1B of human uterine leiomyosarcoma. AB - The cells of tumourigenic line SK-UT-1B combine features characteristic both of normal (diploid karyotype, a low level of polyploid cells, absence of chromosomal marker) and tumour cells (high level of chromosomal instability, high malignancy). We suggest that maintenance of diploid karyotype in this line is controlled via the p53/p21 pathway. We demonstrate that the amount of p53 increases following gamma-irradiation and accumulated p53 protein seems to be functional as p53-luc and p21/Waf-luc reporter plasmids were found to be activated. However, gamma-irradiation-induced increase of p53 was not accompanied by increase of p21/Waf on the protein level. Apparently this is one of the reasons for G1/S and G2/M checkpoint control disruption. The absence of these checkpoints could not prevent the proliferation of cells with intrachromosomal rearrangements. The only effective checkpoint in SK-UT-1B is the p53-dependent M checkpoint, which directed the cells with changed chromosome numbers to apoptosis and therefore strictly guarded the diploidy of the cell population. This indicates that p53 can control the preservation of genetic stability at different levels via different pathways. PMID- 11913955 TI - Saw palmetto berry extract inhibits cell growth and Cox-2 expression in prostatic cancer cells. AB - The cytotoxicity of a commonly used material to alleviate the symptoms of benign prostatic hyperplasia (BPH), Saw Palmetto Berry Extract (SPBE), was examined as neat oil using a set of prostatic cell lines; 267B-1, BRFF-41T and LNCaP. Proliferation of these prostatic derived cell lines is inhibited to different degrees when dosed for 3 days with SPBE. The amount of SPBE required to inhibit 50% growth (IC50) of these cell lines was 20-30 nl equivalents of SPBE per ml of medium for cell lines 267B-1 and BRFF-41T and approximately 10-fold more for the LNCaP cell line. The effect of SPBE dosing on these cell lines is not irreversible, since a 30 min treatment with SPBE at an IC50 concentration does not inhibit their growth. Normal prostate cells were inhibited by 20-25% when grown in the presence of 200 nl SPBE equivalent per ml media. Growth of other non prostatic cancer cell lines, i.e. Jurkat and HT-29, was affected by approx. 50% and 40%, respectively. When LNCaP cells were grown in the presence of dihydrotestosterone and SPBE, the IC50 concentration decreased significantly compared to LNCaP cells grown in the presence of serum and SPBE. Reduced cellular growth after SPBE treatment of these cell lines may relate to decreased expression of Cox-2 and may be due to changes observed in the expression of Bcl 2. Expression of Cox-1 under similar conditions is not affected because of its constitutive expression. Since increased Cox-2 expression is associated with an increased incidence of prostate cancer, and decrease in its expression by SPBE would provide a basis for further investigation of its use against BPH and in prostatic cancer chemoprevention. PMID- 11913956 TI - Role of TGF-beta in the retinoic acid-induced inhibition of proliferation and melanin synthesis in chick retinal pigment epithelial cells in vitro. AB - The purpose of this study was to investigate the effects of all-trans retinoic acid (RA) on the induction of transforming growth factor-beta (TGF-beta) that is concerned with the proliferation and melanin synthesis of chick retinal pigment epithelial (RPE) cells in vitro. Chick RPE cells were cultured in the presence or absence of RA and anti-TGF-beta antibody for 7 days. The effects of RA and pan specific TGF-beta antibody on RPE cell proliferation were assessed by counting the number of cells, and their effects on melanin synthesis were evaluated by measuring the melanin content of the cells. TGF-beta activity in the culture supernatant of RPE cells was measured using CCL-64 cells. RA significantly inhibited RPE cell proliferation and increased melanin synthesis. The addition of pan-specific TGF-beta antibody to the culture blocked the inhibition of RPE cell proliferation and the increased melanin synthesis. RA induced TGF-beta production in the culture supernatant of RPE cells. These findings indicate that RA regulates the proliferation and melanin synthesis of RPE cells via induction of TGF-beta. PMID- 11913957 TI - Cisplatin induces modifications in the development of cell surface patterns of ciliates. AB - Cisplatin [cis-dichlorodiammineplatinum (II)] brings about significant quantitative modifications in the development of cell surface patterns in two unrelated ciliates: Stylonychia and Tetrahymena. Cells cultured in the presence of cisplatin exhibit the formation of supernumerary surface structures in the form of extra cilia/cirri (fused cilia) and other organized ciliary organelles. The metal-induced formation of extra primordia and their differentiation into supernumerary ciliary structures is governed by the normal rules of development. Additional structures are accommodated within the framework of a defined pattern, suggesting the existence of an overall global control of pattern formation. The modified pattern is rectified to its normal state during post-treatment fission cycles, suggesting the role of the cell membrane in correcting developmental errors. PMID- 11913958 TI - Protein kinase C pathway is involved in regulating the secretion of prostatic acid phosphatase in human prostate cancer cells. AB - The stimulated secretion of prostatic acid phosphatase (PAcP) has been known to be a hallmark of androgen action on human prostate epithelial cells for the last five decades. The molecular mechanism of androgen action on PAcP secretion, however, has remained mostly unknown. We investigated the molecular mechanism that promotes PAcP secretion in LNCaP human prostate carcinoma cells which express PAcP and are androgen-responsive. Treatment with 12-o-tetradecanoyl phorbol-13-acetate (TPA), a protein kinase C (PKC) activator, resulted in an increased secretion of PAcP in a dose- and time-dependent fashion. 4Alpha phorbol, a biologically inactive isomer of TPA, had no effect. This TPA stimulation of PAcP secretion was observed 2 h after exposure, while TPA did not have a significant effect on the mRNA level even with a 6 h treatment. A23187 calcium ionophore, known to mobilize cellular calcium which is a co-factor of PKC, also activated PAcP secretion. This TPA stimulation of PAcP secretion was more potent than the conventional stimulating agent 5alpha-dihydrotestosterone (DHT) at the same concentration of 50 nM. Furthermore, the action of TPA and DHT on PAcP secretion was blocked by five different PKC inhibitors. Results also showed that DHT, as well as TPA, could rapidly modulate PKC activity. Therefore, PKC can regulate PAcP secretion, and may also be involved in DHT action on PAcP secretion. PMID- 11913959 TI - New evidence that nuclear import of endogenous beta-catenin is LEF-1 dependent, while LEF-1 independent import of exogenous beta-catenin leads to nuclear abnormalities. AB - The once accepted idea that LEF-1 transports beta-catenin into nuclei has recently been challenged by experiments using exogenous beta-catenin. Here, we investigated the effects of beta-catenin and LEF-1 on nuclear import of beta catenin using different combinations of exogenous and endogenous molecules over longer lengths of time than previously studied. Nuclear beta-catenin is not detectable in corneal fibroblasts and epithelia or NIH 3T3 and MDCK cells. In LEF 1 transfections, we show that the B-box of LEF-1 is required to move cytoplasmic endogenous beta-catenin into the nuclei of such cells, proving that LEF-1 does transport endogenous beta-catenin into nuclei. Moreover, transfection of uveal melanoma cells with B-box deficient LEF-1 inhibits nuclear import of beta-catenin by endogenous LEF-1. However, the movement of overexpressed exogenous beta catenin into nuclei is unaffected by the presence or absence of LEF-1 and forms abnormal nuclear aggregates that are a prelude to subsequent apoptosis. We conclude that nuclear transport of exogenous beta-catenin independently of LEF-1 has questionable physiological significance. PMID- 11913960 TI - Peroxynitrite activates K+-Cl- cotransport in human erythrocytes. AB - Peroxynitrite was found to induce the release of K+ via the Na+/Cl- cotransport system, as do other oxidants. Since peroxynitrite is formed in vivo, its presence could contribute to a pathological dehydration of red blood cells. PMID- 11913961 TI - Distribution and transcription activity of nucleolar DNA in higher plant cells. AB - By using the NAMA-Ur DNA selective staining method, we have observed in situ the location of nucleolar DNA in onion cells and found it at the boundary between fibrillar centres (FC) and dense fibrillar component (DFC) in transcriptionally active nucleolus. We have also used anti-NOR serum, which is identified as the RNA Polymerase I transcription factor (UBF) antibody, to study its reactivity with higher plant cells and demonstrated this factor associated to the DFC but not present at the interior of FC. Finally, by employing anti-DNA/RNA hybrid antibodies, we labeled the transcriptionally active rRNA genes in active nucleolus and testified that at the boundary between FC and DFC. The results provide the evidence that the boundary between FC and DFC is the genuine transcription site of rRNA genes in nucleolus. PMID- 11913962 TI - Effect of vasoactive peptides on Tetrahymena. chemotactic properties of endothelins (ET-1, ET-2, ET-3, fragment 11-21 of ET-1 and big endothelin-1): a short-term inducible signalling mechanism of chemotaxis. AB - The unicellular Tetrahymena is a sensitive model for the study of chemotaxis induced by endothelins. In short-term chemotactic responses, ET-2 and ET-3 were chemorepellent, compared to the referent control (culture medium) and chemoattractant, ET-1. These differences suggest that the change of some aromatic residues in the loop region (residues 5-9) of the ET-1 abrogates its chemoattractant character. The response of Tetrahymena is highly selective, since substitution of two amino acids are enough to cause this alteration in (behavioural) response. Such a change seems to be more important than the loss of the entire first 10 amino acids (in ET-1 fragment 11-21), since after this it acquires some chemoattractive effect of ET-1. Selection with ET-3 rigorously stimulated the cell's responsiveness to the medium, this ability was abolished by the repeated ET-3 treatment. Big endothelin-1 was repellent in all concentrations. These experiments demonstrate the very sensitive discriminating capacity of chemotactic responsiveness at a low level of phylogeny. Chemotactic selection with endothelins underlines the possibility that there may be separate mechanisms responsible for the short-term chemotactic responses and the long lasting effects of chemotactic selection. PMID- 11913963 TI - Effect of neonatal interleukin-6 (IL-6) treatment (hormonal imprinting) on the IL 6 content and localization of the peritoneal, blood and thymic cells of adult rats. A confocal microscopic analysis. AB - Single treatment of newborn rat with human recombinant interleukin-6 (IL-6) durably increases IL-6 content in certain cell types of the adult animal. Peritoneal mast cells contain a high quantity of IL-6, in contrast to controls which contain no IL-6. In addition, many other (mainly lymphatic) cells of the peritoneal fluid contain IL-6. However, IL-6 is present in the control thymocytes, in smaller amounts than in the neonatally IL-6 imprinted cells. In the blood, the number of IL-6 containing cells also increases after imprinting (lymphocytes and developing forms of mast cells). PMID- 11913964 TI - Engineered recombinant enteropeptidase catalytic subunit: effect of N-terminal modification. AB - Enteropeptidase (enterokinase) is a serine protease highly specific for recognition and cleavage of the target sequence of Asp-Asp-Asp-Asp-Lys (D4K). The three-dimensional structure of the enteropeptidase shows that the N-terminal amino acid is buried inside the protein providing molecular interactions necessary to maintain the conformation of the active site. To determine the influence of the N-terminal amino acid of enteropeptidase light chain (EK(L)) on the enzymatic activity, we constructed various mutants including 17 different single amino acid substitutions and three different extensions at the N-terminal end. The mutants of recombinant enteropeptidase (rEK(L)) were expressed in Saccharomyces cerevisiae and secreted into culture medium. Among 20 different mutants tested, the only mutant with the Ile --> Val substitution exhibited significant activity. The kinetic properties of the mutant protein were very similar to those of the wild-type rEK(L). Based on the three-dimensional structure where the N-terminal Ile is oriented into hydrophobic pocket, the results suggest that Val could substitute Ile without affecting the active conformation of the enzyme. The results also explain why all trypsin-like serine proteases carry either Ile or Val at the N-termini and none other amino acid residues are found. Moreover, this finding provides a mental framework for expressing the N-terminally engineered enteropeptidase in Escherichia coli, utilizing the known property of the methionine aminopeptidase that exhibits poor activity toward the N-terminal Met-Ile bond, but offers efficient cleavage of the Met-Val bond. PMID- 11913965 TI - Kinetic peculiarities of human tissue kallikrein: 1--substrate activation in the catalyzed hydrolysis of H-D-valyl-L-leucyl-L-arginine 4-nitroanilide and H-D valyl-L-leucyl-L-lysine 4-nitroanilide; 2--substrate inhibition in the catalyzed hydrolysis of N alpha-p-tosyl-L-arginine methyl ester. AB - Hydrolysis of D-valyl-L-leucyl-L-lysine 4-nitroanilide (1), D-valyl-L-leucyl-L arginine 4-nitroanilide (2), and N alpha-p-tosyl-L-arginine methyl ester (3) by human tissue kallikrein was studied throughout a wide range of substrate concentrations. At low substrate concentrations, the hydrolysis followed Michaelis-Menten kinetics but, at higher substrate concentrations, a deviation from Michaelis-Menten behavior was observed. With the nitroanilides, a significant increase in hydrolysis rates was observed, while with the ester, a significant decrease in hydrolysis rates was observed. The results for substrates (1) and (3) can be accounted for by a model based on the hypothesis that a second substrate molecule binds to the ES complex to produce a more active or an inactive SES complex. The deviation observed for substrate (2) can be explained as a bimolecular reaction between the enzyme-substrate complex and a free substrate molecule. PMID- 11913967 TI - Dissection of the conduit for allosteric control of carbamoyl phosphate synthetase by ornithine. AB - Ornithine is an allosteric activator of carbamoyl phosphate synthetase (CPS) from Escherichia coli. Nine amino acids in the vicinity of the binding sites for ornithine and potassium were mutated to alanine, glutamine, or lysine. The residues E783, T1042, and T1043 were found to be primarily responsible for the binding of ornithine to CPS, while E783 and E892, located within the carbamate domain of the large subunit, were necessary for the transmission of the allosteric signals to the active site. In the K loop for the binding of the monovalent cation potassium, only E761 was crucial for the exhibition of the allosteric effects of ornithine, UMP, and IMP. The mutations H781K and S792K altered significantly the allosteric properties of ornithine, UMP, and IMP, possibly by modifying the conformation of the K-loop structure. Overall, these mutations affected the allosteric properties of ornithine and IMP more than those of UMP. The mutants S792K and D1041A altered the allosteric regulation by ornithine and IMP in a similar way, suggesting common features in the activation mechanism exhibited by these two effectors. PMID- 11913968 TI - A functional analysis of the allosteric nucleotide monophosphate binding site of carbamoyl phosphate synthetase. AB - The catalytic activity of carbamoyl phosphate synthetase (CPS) from Escherichia coli is allosterically regulated by UMP, IMP, and ornithine. Thirteen amino acids within the domain that harbors the overlapping binding sites for IMP and UMP were mutated to alanine and characterized. The four residues that interact directly with the phosphate moiety of IMP in the X-ray crystal structure (K954, T974, T977, and K993) were shown to have the greatest impact on the dissociation constants for the binding of IMP and UMP and the associated allosteric effects on the kinetic constants of CPS. Of the four residues that interact with the ribose moiety of IMP (S948, N1015, T1017, and S1026), S1026 was shown to be more important for the binding of IMP than UMP. Five residues (V994, I1001, D1025, V1028, and I1029) were mutated in the region of the allosteric domain that surrounds the hypoxanthine ring of IMP. With the exception of V994A, these mutations had a modest influence on the binding and subsequent allosteric effects by UMP and IMP. PMID- 11913966 TI - Cellular oxidant stress and advanced glycation endproducts of albumin: caveats of the dichlorofluorescein assay. AB - In order to understand the mechanism by which advanced glycation endproducts (AGEs) elicit oxidative stress, macrophage-like RAW264.7 cells were exposed to various AGE-albumins, and oxidant stress was estimated from the fluorescence of oxidized dichlorofluorescein using the microtiter plate assay. Strongest fluorescence was observed with methylglyoxal modified albumin (MGO-BSA) compared with native albumin. Similar effects that were prevented by arginine coincubation were seen with phenylglyoxal-BSA. MGO-BSA had increased affinity for Cu(2+) and Ca(2+), but was conformationally similar to native albumin. Surprisingly, the mere addition of unmodified albumin to cells suppressed the fluorescence of oxidized DCF. While, several site-directed mutants of human serum albumin (HSA), including C34S and recombinant domains II and III retained fluorescence suppressing activity, proteolytic digests, recombinant domain I, and several nonalbumin proteins failed to suppress. Kinetic and ANS binding studies suggested albumin quenches DCF fluorescence by binding to hydrophobic pockets in domains II and III and that MGO-BSA is less hydrophobic than BSA. Finally, BSA also prevented H(2)O(2) catalyzed DCF fluorescence more potently than MGO-BSA. These studies reveal important caveats of the widely used dichlorofluorescein assay and suggest methods other than the microtiter plate assay are needed to accurately assess cellular oxidant stress in presence of native or modified albumin. PMID- 11913970 TI - Xanthine oxidase and aldehyde oxidase: a simple procedure for the simultaneous purification from rat liver. AB - Aldehyde oxidase (AO) and xanthine oxidase (XO) are cytosolic enzymes that have been involved in some pathological conditions and play an important role in the biotransformation of drugs and xenobiotics. The increasing interest in these enzymes demands for a simple and rapid procedure for their purification. This paper describes for the first time a method that allows simultaneous purification of both enzymes from the same batch of rat livers. It involves few steps, is reproducible and offers high enzyme yields with high specific activities. The rat liver homogenate was fractionated by heat denaturation and by ammonium sulphate precipitation to give a crude extract containing both enzymes. This extract was chromatographed on an Hydroxyapatite column that completely separated AO from XO. Further purification of XO by anion exchange chromatography on a Q-Sepharose Fast Flow column resulted in a highly purified (1200-fold) preparation, with a specific activity of 3.64 U/mg and with a 20% yield. AO was purified about 1000 fold at a yield of 15%, with a specific activity of 3.48 U/mg, by affinity chromatography on Benzamidine-Sepharose 6B. The purified enzymes gave single bands of approximately 300 kDa on a polyacrylamide gel gradient electrophoresis and displayed the characteristic absorption spectra of highly purified enzymes. PMID- 11913969 TI - Inhibition of transthyretin amyloid fibril formation by 2,4-dinitrophenol through tetramer stabilization. AB - Transthyretin (TTR), a homotetrameric thyroxine transport protein found in the plasma and cerebrospinal fluid, circulates normally as a innocuous soluble protein. In some individuals, TTR polymerizes to form insoluble amyloid fibrils. TTR amyloid fibril formation and deposition have been associated with several diseases like familial amyloid polyneuropathy and senile systemic amyloidosis. Inhibition of the fibril formation is considered a potential strategy for the therapeutic intervention. The effect of small water-soluble, hydrophobic ligand 2,4-dinitrophenol (2,4-DNP) on TTR amyloid formation has been tested. 2,4-DNP binds to TTR both at acidic and physiological pH, as shown by the quenching of TTR intrinsic fluorescence. Interestingly, 2,4-DNP not only binds to TTR at acidic pH but also inhibits amyloid fibril formation as shown by the light scattering and Congo red-binding assay. Inhibition of fibril formation by 2,4-DNP appears to be through the stabilization of TTR tetramer upon binding to the protein, which includes active site. These findings may have implications for the development of mechanism based small molecular weight compounds as therapeutic agents for the prevention/inhibition of the amyloid diseases. PMID- 11913971 TI - Molecular and regulatory properties of leucoplast pyruvate kinase from Brassica napus (rapeseed) suspension cells. AB - Plastidic pyruvate kinase (PK(p)) from Brassica napus suspension cells was purified 431-fold to a final specific activity of 28 micromol phosphoenolpyruvate (PEP) utilized/min/mg protein. SDS-PAGE, immunoblot and gel filtration analyses indicated that this PK(p) exists as a 380-kDa heterohexamer composed of equal proportions of 64- (alpha-subunit) and 58-kDa (beta-subunit) polypeptides. The N terminal sequence of the PK(p) alpha- and beta-subunits exhibited maximal identity with the corresponding regions deduced from putative PK genes of Arabidopsis thaliana and Methylobacterium extorquens, respectively. B. napus PK(p) displayed a sharp pH optimum of pH 8.0, and hyperbolic saturation kinetics with PEP and ADP (K(m) = 0.052 and 0.14 mM, respectively). 6-Phosphogluconate functioned as an activator (K(a) = 0.12 mM) by increasing V(max) by approximately 35% while decreasing the K(m)(PEP) and K(m)(ADP) values by 40 and 50%, respectively. 2-Oxoglutarate and oxalate were the most effective inhibitors (I(50) = 8.3 and 0.23 mM, respectively). A model is presented which highlights the role of 6-phosphogluconate in coordinating stromal NADPH and ATP production for anabolic processes of B. napus leucoplasts. PMID- 11913972 TI - Heterologous expression of an endogenous rat cytochrome b(5)/cytochrome b(5) reductase fusion protein: identification of histidines 62 and 85 as the heme axial ligands. AB - The gene coding for expression of an endogenous soluble fusion protein comprising a b-type cytochrome-containing domain and a FAD-containing domain has been cloned from rat liver mRNA. The 1461-bp hemoflavoprotein gene corresponded to a protein of 493 residues with the heme- and FAD-containing domains comprising the amino and carboxy termini of the protein, respectively. Sequence analysis indicated the heme and flavin domains were directly analogous to the corresponding domains in microsomal cytochrome b(5) (cb5) and cytochrome b(5) reductase (cb5r), respectively. The full-length fusion protein was purified to homogeneity and demonstrated to contain both heme and FAD prosthetic groups by spectroscopic analyses and MALDI-TOF mass spectrometry. The cb5/cb5r fusion protein was able to utilize both NADPH and NADH as reductants and exhibited both NADPH:ferricyanide (k(cat) = 21.7 s(-1), K(NADPH)(m) = 1 microM. K(FeCN6)(m) = 8 microM) and NADPH:cytochrome c (k(cat) = 8.3 s(-1), K(NADPH)(m) = 1 microM. K(cyt c)(m) = 7 microM) reductase activities with a preference for NADPH as the reduced pyridine nucleotide substrate. NADPH-reduction was stereospecific for transfer of the 4R proton and involved a hydride transfer mechanism with a kinetic isotope effect of 3.1 for NADPH/NADPD. Site-directed mutagenesis was used to examine the role of two conserved histidine residues, H62 and H85, in the heme domain segment. Substitution of either residue by alanine or methionine resulted in the production of simple flavoproteins that were effectively devoid of both heme and NAD(P)H:cytochrome c reductase activity while retaining NAD(P)H:ferricyanide activity, confirming that the former activity required a functional heme domain. These results have demonstrated that the rat cb5/cb5r fusion protein is homologous to the human variant and has identified the heme and FAD as the sites of interaction with cytochrome c and ferricyanide, respectively. Mutagenesis has confirmed the identity of both axial heme ligands which are equivalent to the corresponding residues in microsomal cytochrome b(5). PMID- 11913973 TI - A novel in vitro assay for deubiquitination of I kappa B alpha. AB - The ubiquitin-proteasome pathway (UPP) controls a wide range of signal transduction cascades by targeting key regulatory proteins for 26S proteasome mediated degradation. Several observations suggest that protein deubiquitination may modulate this process; however, few experiments have been performed to test this idea. An excellent model system for studying the regulatory role of the UPP is signal transduction via the nuclear factor-kappa B (NF-kappa B) family of transcription factors. The principal inhibitor of NF-kappa B, I kappa B alpha, is polyubiquitinated and degraded in response to diverse stimuli. In this study, we sought to determine whether I kappa B alpha deubiquitination also occurs. We established an in vitro deubiquitination assay using polyubiquitinated I kappa B alpha as the substrate. Our data provide evidence of an I kappa B alpha-directed deubiquitinating activity present in lysates of several cell lines. This activity was inhibited by ubiquitin aldehyde, a specific inhibitor of deubiquitinating enzymes, as well as by alkylating reagents or heat, but was unaffected by the inhibition of several other classes of proteases. Cell lysates and the deubiquitinating enzyme, UCH-L3, hydrolyzed ubiquitin 7-amido-4-methylcoumarin, a model substrate for assaying deubiquitinating activities. However, UCH-L3 had no detectable activity toward ubiquitinated I kappa B alpha, thus suggesting a degree of enzymatic specificity in the deubiquitination of I kappa B alpha. This assay will be useful for the study of I kappa B alpha deubiquitination. Moreover, this assay can be adapted to monitor the deubiquitination of other proteins modified by ubiquitin conjugation. PMID- 11913974 TI - Translation and phosphorylation of wheat germ lysate: phosphorylation of wheat germ initiation factor 2 by casein kinase II and in N-ethylmaleimide-treated lysates. AB - Previously, we observed that N-ethylmaleimide (NEM), a thiol-alkylating agent, was found to stimulate the phosphorylation of several proteins in translating wheat germ (WG) lysates, including the phosphorylation of alpha, the p41-42 doublet subunit, and beta, the p36 subunit, of the WG initiation factor 2 (eIF2). We find now that NEM increases phosphorylation of several proteins significantly in lysates which are moderate or low in their translation compared to optimally active lysates. Heat treatment, which stimulates oxidation of protein sulfhydryls, decreases the translation and phosphorylation ability of WG lysates. The decrease in phosphorylation, but not translation, that occurs in heat-treated lysates is prevented very efficiently by NEM and partially by reducing agents such as dithiothreitol (DTT) and GSH. DTT prevents, however, completely the loss of sulfhydryl content of heat-treated WG lysates and does not at all prevent heat induced inhibition of translation. In contrast, DTT prevents completely the diamide-induced translational inhibition and also the loss of sulfhydryl content. These findings therefore suggest that in addition to the maintenance of sulfhydryl groups, heat-labile proteins and their interactions with other proteins play an important role in overall translation and phosphorylation. It is also observed here that heat treatment stimulates the phosphorylation of rabbit reticulocyte eIF2 alpha but not the alpha subunit (p41-42 doublet) of WG eIF2. A phosphospecific anti-eIF2 alpha antibody recognizes the WG eIF2 alpha(P) that is phosphorylated by an authentic eIF2 alpha kinase such as double-stranded RNA dependent protein kinase, but it is unable to recognize the eIF2 alpha that is phosphorylated in NEM-treated lysates. These findings therefore suggest that phosphorylation of WG eIF2 alpha in NEM-treated lysates occurs on a site different from the serine 51 residue that is phosphorylated by authentic eIF2 alpha kinases. In addition, it also suggests that WG eIF2 alpha, unlike reticulocyte eIF2 alpha, is phosphorylated by eIF2 alpha kinases and also by other kinases. Consistent with this idea, it has been observed here that casein kinase II (CKII) phosphorylates WG eIF2 alpha and the phosphorylation is enhanced by NEM in vitro and in lysates. The phosphopeptide analysis suggests that WG eIF2 alpha has separate phosphorylation sites for CKII and heme-regulated eIF2 alpha kinase (a well-characterized mammalian eIF2 alpha kinase), and NEM-induced phosphorylation in WG lysates resembles CKII-mediated phosphorylation. PMID- 11913977 TI - Redox control in heme proteins: electrostatic substitution in the active site of leghemoglobin. AB - The effects of electrostatic substitutions on the spectroscopic, ligand binding, and redox properties of the heme in leghemoglobin have been examined by replacement of the proximal leucine 88 residue with an aspartic acid residue (Leu88Asp). Electronic and resonance Raman spectra of the ferric derivative of Leu88Asp indicate a mixture of 6-coordinate, high-spin and 6-coordinate, low-spin hemes, analogous to that observed in the recombinant wild-type protein (rLb). At alkaline pH, formation of hydroxide-bound heme is indicated for Leu88Asp; the pK(a) for this transition (8.7 +/- 0.2, micro = 0.10 M, 25.0 degrees C) is 0.4 pH units higher than for rLb. Equilibrium dissociation constants (sodium phosphate, pH 7.0, micro = 0.10 M, 25.0 +/- 0.1 degrees C) for binding of anionic ligands (N(-)(3), nicotinate) to Leu88Asp are higher (K(d,nicotinate) = 6.8 +/- 0.2 microM; K(d,azide) = 33 +/- 0.6 microM) than the corresponding values for rLb (K(d,nicotinate) = 1.4 +/- 0.3 microM (pH 5.5, micro = 0.10 M, 25.0 +/- 0.1 degrees C); K(d,azide) = 4.8 +/- 0.2 microM). Resonance Raman spectra (sodium phosphate, pH 7.0, micro = 0.10 M) for the ferrous derivatives of Leu88Asp and rLb exhibit a strong nu(Fe-His) stretching frequency at 223 cm(-1) in both cases, indicating that the hydrogen bonding structure on the proximal side is not substantially altered in the variant. The reduction potential of Leu88Asp is -14 +/- 2 mV vs standard hydrogen electrode (SHE) (25.0 degrees C, micro = 0.10 M, pH 7.0), a decrease of 35 mV over the corresponding value for the wild-type protein under the same conditions (21 +/- 3 mV vs SHE). An assessment of these data in terms of electrostatic and hydrogen bonding considerations is presented. PMID- 11913975 TI - Tom34 unlike Tom20 does not interact with the leader sequences of mitochondrial precursor proteins. AB - Tom20 and Tom34 are mammalian liver proteins previously identified by others to be components of the mitochondrial import translocation apparatus. It has been shown that Tom20 interacts with the leader sequence of nuclear coded matrix space precursor proteins. Here we show with recombinantly expressed Tom proteins that Tom34 binds the mature portion of the precursor and not the leader. Both these Tom proteins inhibited the import of newly translated precursor of aldehyde dehydrogenase in an in vitro assay. Only Tom20 inhibited the import of a fusion protein of the leader of aldehyde dehydrogenase attached to dihydrofolate reductase. Antibodies against Tom20 coprecipitated both the precursor of aldehyde dehydrogenase (pALDH) and of dihydrofolate reductase (pA-DHFR). Antibodies against Tom34 interacted only when the mature portion of aldehyde dehydrogenase was present. Similar import inhibition patterns were found when other precursor and chimeric constructs we investigated. When Tom34-green fluorescence protein was transfected to HeLa cells it was observed that Tom34 was found through out the cell. It is concluded from our observation that Tom34 is a cytosolic protein, whose role appeared to be to interact with mature portion of some preproteins and may keep them in an unfolded, import compatible state. PMID- 11913976 TI - Yeast two-hybrid screening identifies binding partners of human Tom34 that have ATPase activity and form a complex with Tom34 in the cytosol. AB - In the accompany paper (Mukhopadhyay, A., Avramova, L. V. and Weiner, H., Arch. Biochem. Biophys.), it was shown that Tom34, a previously proposed putative translocase of the mitochondrial outer membrane, binds to the mature region of a precursor protein and appears to be a cytosol protein. Here Tom34 was used as bait in a yeast two-hybrid screening to search for its potential binding partners. Two of the identified proteins were the ATPase-related valosin containing protein (VCP) and the lysosomal H(+)-transporting ATPase member M (ATP6M). Tom34 was found primarily in the cytosol while VCP and ATP6M were found in the cytosol as well as in nonmitochondrial organelles. Tom34 formed a approximately 400-kDa complex with them in the cytosol. Tom34 was found to possess a weak ATPase activity that did not change when associated with VCP. The tetratricopeptide repeat (TPR) motif region of Tom34 (residue 201-256) was responsible for binding to the other proteins. Tom34 appears not to be a member of the mitochondrial outer membrane translocase family but might function as a chaperone-like protein during protein translocation. PMID- 11913979 TI - Biosynthesis of marine natural products: isolation and characterization of a myrcene synthase from cultured tissues of the marine red alga Ochtodes secundiramea. AB - The acyclic monoterpene myrcene is the likely progenitor of the unusual cytotoxic halogenated monoterpenes that are found in marine algae and that function as feeding deterrents to herbivores. Myrcene synthase was isolated from suspension cultures of the marine red alga Ochtodes secundiramea, representing the first enzyme of this type from a marine organism. The algal myrcene synthase produces exclusively myrcene from the natural substrate geranyl diphosphate (GDP), utilizes Mg(+2) as the required divalent metal ion cofactor, has a molecular mass of about 69 kDa, and exhibits a pH optimum near 7.2. These features are similar to those of monoterpene synthases from terrestrial organisms. When incubated with neryl diphosphate (the cis-isomer of GDP), the O. secundiramea myrcene synthase produces the cyclic monoterpene limonene, whereas incubation with (+/-)linalyl diphosphate (the tertiary allylic isomer of geranyl diphosphate) yields both acyclic and cyclic monoterpenes. These results suggest that the enzyme is incapable of isomerizing geranyl diphosphate to linalyl diphosphate, a feature common to all monoterpene cyclases from terrestrial sources. The limited catalytic capability of the myrcene synthase may reflect the ancient evolutionary origin of the producing organism. The ability to assay this enzyme in cultured algae, grown under strictly defined conditions, provides an unparalleled opportunity to delineate factors eliciting the biosynthesis of this class of secondary metabolites, to investigate the metabolic pathway leading to the halogenated monoterpenes, and to determine their role in the chemical ecology of marine algae. PMID- 11913978 TI - Regulation of calbindin-D9k expression by 1,25-dihydroxyvitamin D(3) and parathyroid hormone in mouse primary renal tubular cells. AB - Calbindin (CaBP)-D9k is a major vitamin D target gene involved in calcium homeostasis. However, studies on the molecular mechanisms of CaBP-D9k gene regulation have been hampered by the lack of an appropriate cell culture system. In the present study, we used mouse primary renal tubular cell (PRTC) cultures to investigate the regulation of CaBP-D9k expression by 1,25(OH)(2)D(3). Both CaBP D9k mRNA and protein were highly induced by 1,25(OH)(2)D(3) in a time- and dose dependent manner in PRTCs, and new RNA and protein synthesis was required for the induction. Transfection of VDR(-/-) cells derived from VDR null mice with human VDR restored the induction of CaBP-D9k expression by 1,25(OH)(2)D(3), confirming the requirement of VDR for CaBP-D9k expression. Treatment of the PRTCs with 1,25(OH)(2)D(3) also increased VDR protein abundance, suggesting that enhanced VDR transactivation is involved in the CaBP-D9k up-regulation. Moreover, PTH had a synergistic effect on the 1,25(OH)(2)D(3) induction of CaBP-D9k. These data demonstrate that CaBP-D9k is highly regulated by 1,25(OH)(2)D(3) and PTH in mouse PRTCs, which provides a suitable in vitro system for further investigating the molecular mechanisms involved in CaBP-D9k gene regulation. PMID- 11913980 TI - Arachidonic acid converts the glutathione depletion-induced apoptosis to necrosis by promoting lipid peroxidation and reducing caspase-3 activity in rat glioma cells. AB - Intracellular glutathione (GSH) depletion induced by buthionine sulfoximine (BSO) caused cell death that seemed to be apoptosis in C6 rat glioma cells. Arachidonic acid (AA) promoted BSO-induced cell death by accumulating reactive oxygen species (ROS) or hydroperoxides. AA inhibited caspase-3 activation and internucleosomal DNA fragmentation during the BSO-induced GSH depletion. Furthermore, AA reduced intracellular ATP content, induced dysfunction of mitochondrial membrane and enhanced 8-hydroxy-2'-deoxyguanosine (8-OH-dG) production. There was significant increase of 12-lipoxygenase activity in the presence of AA under the BSO-induced GSH depletion in C6 cells. These results suggest that AA promotes cell death by changing to necrosis from apoptosis through lipid peroxidation initiated by lipid hydroperoxides produced by 12-lipoxygenase under the GSH depletion in C6 cells. Some ROS such as hydroperoxide produced by unknown pathway make hydroxy radicals and induce 8-OH-dG formation in the cells. The conversion of apoptosis to necrosis may be a possible event under GSH depleted conditions. PMID- 11913981 TI - Both the dimerization and immunochemical properties of E-cadherin EC1 domain depend on Trp(156) residue. AB - Using site-directed mutagenesis, we show in this paper that the adhesive interface detected in cadherin crystals is unlikely to mediate adhesive interaction between myc- and flag-tagged E-cadherin molecules in human A-431 cells. We also found that a critical residue within this interface, His(233), is part of the epitope for mAb SHE78-7. This epitope was accessible to the antibody in the adhesive E-cadherin dimers, which is consistent with uninvolvement of the site containing His(233) in cell-cell adhesion. However, both the adhesive dimerization and the integrity of the SHE78-7 epitope depended on the same intramolecular interaction between Trp(156) and its hydrophobic pocket. Our data suggest that this interaction may have an important regulatory function in controlling the surface topology of the NH(2)-terminal domain of E-cadherin. PMID- 11913982 TI - Pathophysiology of platelet destruction in immune (idiopathic) thrombocytopenic purpura. AB - The mechanism of platelet destruction in immune (idiopathic) thrombocytopenic purpura (ITP) is thought to involve production of autoantibody to platelet surface antigens. Once coated with antibody, circulating platelets undergo sequestration via interaction with Fc receptors of macrophages in the reticuloendothelial system. A number of questions remain about the mechanism of platelet destruction in this disease: 1) What is the nature of the stimulus to the immune system that generates antiplatelet antibodies? 2) What is the role of interactions between T-helper lymphocytes and antigen-presenting cells in ITP? 3) What role, if any, is played by the targeting of single or multiple platelet surface glycoproteins by the autoimmune response? 4) Is the site of platelet destruction, intravascular or extravascular, or the state of activation of platelets important in the destruction of platelets? PMID- 11913983 TI - Humanized mouse models of FcR clearance in immune platelet disorders. AB - Transgenic mouse lines expressing physiologic levels of human platelet Fc receptor (FcR) for IgG, Fc gamma RIIA, on platelets and macrophages were generated. Anti-CD9 antibody activated platelets of Fc gamma RIIA transgenic mice and, following injection in vivo, caused rapid and severe thrombocytopenia compared with nonactivating antiplatelet antibody. Anti-CD9 injected in Fc gamma RIIA transgenic mice crossed with FcR gamma-chain knockout (gamma-KO) mice caused thrombosis and shock in all mice, and death in 16 of 18 mice. Histologic examination of lung vasculature of anti-CD-treated Fc gamma RIIA transgenic x gamma-KO mice showed extensive platelet-fibrin thrombi. Taken together, these observations suggest that in Fc gamma RIIA transgenic x gamma-KO mice there is an important interplay of intravascular platelet activation and splenic clearance. Reduction of splenic clearance surgically or functionally also facilitated anti CD-9-mediated shock in Fc gamma RIIA transgenic mice. Thus, antibodies that activate platelets in an Fc gamma RIIA-dependent manner lead to thrombosis, shock, and death. These mouse model findings have implications for human immune mediated thrombocytopenic disorders. Genetic factors may be important in the interindividual variability seen clinically, and with nonactivating platelet antibody the spleen is largely responsible for the thrombocytopenia. This is likely the case in typical idiopathic thrombocytopenic purpura (ITP). For several other immune thrombocytopenic disorders, the spleen probably plays a second, protective role in removing antibody-coated platelets from the circulation. PMID- 11913984 TI - Pathophysiology and thrombokinetics in autoimmune thrombocytopenia. AB - Studies that have measured platelet survival by autologous platelet labeling with (111)In and (51)Cr have differed in their results. Although all studies have revealed a significant decrease in platelet life span, the rates of platelets entering the circulation, a calculated and inferred determination, have been found to be moderately decreased to as much as five times normal. Several mechanisms have been proposed to explain an apparent decrease in platelet production, including a true decrease due to damage to the megakaryocytes by autoantibody, versus a decrease only in 'effective' production due to intramedullary destruction by the reticuloendothelial system. Recently, the identification of the cytokine, thrombopoietin, has allowed the evaluation of another aspect of the pathophysiology of thrombocytopenic states. Megakaryocyte growth factor levels are increased 10 to 20 times in patients who are thrombocytopenic due to chemotherapy or aplastic anemia, but may be decreased, normal, or only modestly raised in patients with immune platelet destruction. Autologous platelet survival measurements, prior to and while on therapy with a stable platelet count, reveal that removal of part of the reticuloendothelial system with splenectomy leads to increased platelet survival and platelet number. Similar studies reveal that corticosteroid treatment for immune thrombocytopenic purpura (ITP) effectively increases the rate of platelet production but does not change platelet survival. It may be that other therapies are effective by a combination of these mechanisms. Stimulation of thrombopoietin production or administering exogenous cytokine may have a role to play in future management of patients with ITP. PMID- 11913985 TI - Immune pathophysiology of autoimmune thrombocytopenic purpura. AB - Chronic autoimmune thrombocytopenic purpura (AITP) is an immune-mediated, bleeding disorder in which platelets are opsonized by autoantibodies and prematurely destroyed by phagocytic cells in the reticuloendothelial system. It is classed as an organ-specific autoimmune disease primarily mediated by immunoglobulin G (IgG) autoantibodies and its etiology appears to be similar to that observed for other organ-specific autoimmune diseases. Th1 cells are important in the process, and the costimulation of Th1 cells and B cells takes place in a cytokine milieu that is reminiscent of a proinflammatory process. Chronic AITP has classically been treated with nonspecific, immunosuppressive regimens (e.g., steroids). One of the most significant developments in the treatment of AITP in the last 20 years has been the use of intravenous immunoglobulin (IVIg) and anti-D preparations. These treatments confer benefit to patients with AITP by significantly raising platelet counts. Despite this, their exact mechanisms of action remain elusive. This review focuses on cell-mediated and cytokine abnormalities within AITP, and presents data related to the mechanism of action of anti-D. PMID- 11913986 TI - Pathophysiology of immune thrombocytopenic purpura. AB - In 1951, the young hematologist in training, Dr. William Harrington, infused himself with plasma from a patient with immune thrombocytopenic purpura (ITP). He rapidly developed severe, but transient, thrombocytopenia and was at risk for serious hemorrhage. Thus, the humoral autoimmune cause of ITP was established. Since 1953, when Dr. Harrington's in vivo studies ended, in vitro investigations have aimed to determine the molecular and cellular details of immune-mediated platelet destruction. PMID- 11913987 TI - Controversies in the treatment of pediatric immune thrombocytopenias. AB - Although pediatric immune thrombocytopenia (ITP) is common, there is no consensus on the optimal approach to therapy. Childhood ITP differs in its clinical course and trigger from adult immune thrombocytopenic purpura. There appear to be two clinical phenotypes among children with ITP: children with polyclonal autoantibody production triggered by an external exposure such as infection, and children with coexistent immune deficiency or dysregulation on a congenital or acquired basis. Treatment implications exist for each group. The first may be best managed by observation and conservative measures; for the latter, treatment could include normal intravenous gammaglobulin concentrate, anti-blood group D antigen (anti-Rh factor), or steroids. Early recognition of the thrombocytopenic child with immune dysfunction versus the normal child with a polyclonal response to a particular environmental antigen will result in better prognosis for both by selecting the appropriate therapy and minimizing long-term side effects. PMID- 11913988 TI - Acute immune (idiopathic) thrombocytopenic purpura in childhood. AB - Acute immune (idiopathic) thrombocytopenic purpura (ITP) in childhood is most commonly a self-limiting condition with unexplained onset and resolution. In cases of severe thrombocytopenia, or situations where the condition persists beyond 6 months, treatment may be required to minimize the danger of life threatening intracranial hemorrhage. Nonsurgical treatment options include corticosteroids, intravenous gammaglobulin (i.v.Ig), or anti-D. Specific indications, benefits, and limitations of these modalities are discussed, with recommendations for future directions in therapy. PMID- 11913989 TI - Childhood chronic immune thrombocytopenic purpura (ITP). AB - Chronic immune thrombocytopenic purpura (ITP) develops in approximately 20% of children with acute ITP. Treatment requires differentiating the truly idiopathic state from thrombocytopenias associated with other disorders. Treatment is recommended in cases where platelet counts drop below 20 x 10(9)/L. Therapeutic options include corticosteroids, intravenous immunoglobulin TG(i.v.Ig), and anti D, as well as splenectomy. The benefits and disadvantages of each modality are reviewed, as well as the management of postsplenectomy failures. PMID- 11913990 TI - Childhood immune thrombocytopenic purpura. AB - Childhood immune thrombocytopenic purpura (ITP) is acute and generally seasonal in nature, suggesting that infectious or environmental agents may trigger the immune response to produce platelet-reactive autoantibodies 4 to 8 weeks following an infection. In general, the patient is well apart from the diffuse bruising and petechiae indicative of a profound thrombocytopenia. Over a period of 6 months, the thrombocytopenia resolves in approximately 85% of children, while the remaining 15% with persistent platelet consumption are designated as chronic ITP patients. The peak age of acute ITP is 2 to 5 years of age, a period when children experience the greatest frequency of viral infections. Children with the chronic form of ITP mirror the adult phenotype, in that females predominate, and there is no seasonal fluctuation of the disease. Evidence from our laboratory suggests that the activated platelet itself may play a role in perpetuating autoantibody production and immune dysregulation associated with ITP. Current data on lymphocyte studies and cytokine alterations noted in response to the variety of regimens used in children with ITP suggest that acute ITP is accompanied by autoantibodies to GPIb and a cytokine profile that is proinflammatory in nature. Early recognition of the immune dysregulation driving acute versus chronic ITP will distinguish those children who might benefit from immunotherapy versus those who will recover without therapeutic intervention. PMID- 11913991 TI - Novel approaches to refractory immune thrombocytopenic purpura. AB - Chronic immune thrombocytopenic purpura (ITP) is an organ-specific autoimmune bleeding disorder in which autoantibodies are directed against the individual's own platelets, resulting in increased Fc-mediated platelet destruction by macrophages in the reticuloendothelial system. Although ITP is primarily mediated by IgG autoantibodies, the production of these autoantibodies is regulated by the influence of T lymphocytes and antigen-presenting cells (APC). There is evidence that enhanced T-helper cell/APC interactions in patients with ITP may play an integral role in IgG antiplatelet autoantibody production. New therapies may improve platelet production, decrease platelet antibody production, and decrease monocyte function and/or B-cell and T-cell activities. Understanding these cellular immune responses in ITP may lead to the development of more specific immunoregulatory therapies for the management of this disease. PMID- 11913992 TI - Initial management of adults with idiopathic (immune) thrombocytopenic purpura. AB - Since idiopathic (immune) thrombocytopenic purpura (ITP) in adults is usually a chronic condition with few spontaneous remissions, the goal of treatment is not cure, but to maintain a hemostatically safe platelet level. The indication for treatment should be based not merely on platelet counts, but also clinical indices of bleeding. Although most patients show good initial response to prednisone, the side effects of steroids limit this treatment. Currently, long term management usually involves splenectomy. Since splenectomy has surgical risks and may also predispose the patient to sepsis, a clinical trial using anti D (WinRho-SDR) has been performed to determine whether this treatment can safely delay or avoid the need for surgery. The use of WinRho may also reveal the occurrence of spontaneous remissions, a previously unrecognized subgroup of adults with chronic ITP. PMID- 11913993 TI - Role of splenectomy in immune (idiopathic) thrombocytopenic purpura. AB - Idiopathic thrombocytopenic purpura (ITP) is an immune disorder, which causes an acute or chronic thrombocytopenia, and may result in potentially life-threatening hemorrhage. Splenectomy is one of the treatment options that needs to be weighed in the treatment of ITP, particularly in cases that have shown response failure to medical modalities such as prednisone, i.v.Ig, or anti-D globulin therapy. Although most studies demonstrate good early response following splenectomy, the long-term outcome is less favorable. Furthermore, other negative factors, such as rendering the patient ineligible for anti-D globulin or oral tolerance therapy and vulnerable to possible life-threatening sepsis, must be weighed prior to splenectomy. PMID- 11913994 TI - Idiopathic thrombocytopenic purpura complicating pregnancy. AB - Idiopathic thrombocytopenic purpura (ITP) is a condition that often develops in young women, and, consequently, physicians will frequently manage pregnant patients with this disorder. The lack of prospective trials investigating ITP in pregnancy has meant that physicians frequently rely on personal experience and anecdotal evidence to manage these patients. However, recent, large, prospective and retrospective studies have clarified several important aspects of ITP in pregnancy. First, ITP should be distinguished from incidental thrombocytopenia of pregnancy, a high frequency and benign, mild drop in the platelet count that occurs late in pregnancy. Second, although the platelet count often drops in pregnant ITP patients, serious morbidity or mortality is distinctly uncommon for the mothers. The overall frequency of thrombocytopenia in the infant is quite low, and about 5% of infants will have a birth platelet count less than 20 x 10(9)/L and about 1% will have significant bleeding complications. PMID- 11913995 TI - Clinical management of adult ITP prior to splenectomy: a perspective. AB - A philosophy of management of adult immune thrombocytopenic purpura (ITP) prior to splenectomy is presented. The initial action is to determine whether the condition is hyperacute, acute, or chronic. In symptomatic cases, initial remission usually requires steroids and the administration of intravenous immunoglobulin (i.v.Ig), followed by platelet transfusion if the patient is actively hemorrhaging. Once initial remission is achieved, a rapid reduction to minimal maintenance therapy should be made. The options include steroids, immunotherapy, and chemotherapy. Adjustments to maintenance therapy must be made with the understanding that response rates vary for each agent. PMID- 11913996 TI - Classical management of refractory adult immune (idiopathic) thrombocytopenic purpura. AB - Treatment of chronic immune (idiopathic) thrombocytopenic purpura with corticosteroids and/or splenectomy results in safe platelet counts in over 70% of patients without additional treatment. Therapy of patients who are refractory to these two treatments may be difficult. The treatment approach to refractory ITP patients, described in this report, is arbitrarily divided into four levels: levels 1 through 3 represent treatments with increasing side effects; level 4 therapy may be tried when the others have failed. Patients undergoing these treatments may require concomitant intravenous gammaglobulin, high-dose corticosteroids or platelets, to maintain the platelet count in the setting of mucosal bleeding or severe thrombocytopenia. PMID- 11913997 TI - The potential role of thrombopoietin in idiopathic thrombocytopenic purpura. AB - Thrombopoietin (TPO) plays a central role in the pathogenesis of idiopathic thrombocytopenic purpura (ITP), as it does in other immune-mediated thrombocytopenias. Because TPO is bound and internalized by platelets, it is destroyed together with platelets at an accelerated rate in the macrophage system. Because the spleen acts as a TPO sink, compensation of the decreased platelet count by an increased production in the bone marrow is insufficient. This aspect of ITP, as well as the use of TPO in the treatment of ITP, will be the focus of this article. PMID- 11913998 TI - Hematologic complications of primary immune deficiencies. AB - Primary immune deficiencies have an estimated overall incidence of 1 in 10,000 individuals. These disorders are diverse, depending on the specific immune functions involved, and lead to chronic or recurrent infections, inflammatory conditions, and a variety of autoimmune diseases. The most common autoimmune disorder is immune thrombocytopenic purpura (ITP), followed by autoimmune hemolytic anemia (AHA). While cytopenias are common in all the congenital immune diseases, they are particularly common in the antibody defects, common variable immunodeficiency and selective immunoglobulin A deficiency. In common variable immunodeficiency, ITP occurred in 7.6% of the patients and AHA in 4.8%. Treatment options include corticosteroids, intravenous immunoglobulin (i.v.Ig), anti-D, and splenectomy. Although the association between cytopenias and congenital immune deficiency is unclear, defects in T-cell regulation, cytokine defects, abnormal apoptosis, and abnormal production of immunoglobulins with autoimmune features are potential mechanisms. PMID- 11913999 TI - Pharmacoeconomics of therapy for ITP: steroids, i.v.Ig, anti-D, and splenectomy. AB - Factors that influence the choice of therapeutic option in treating idiopathic thrombocytopenic purpura (ITP) include cost, toxicity, and efficacy of the agent. Today the most commonly used agents for treating ITP include glucocorticoids, intravenous immunoglobulin (i.v.Ig), anti-D, and splenectomy. Splenectomy cannot be avoided in some instances, but i.v.Ig, and anti-D offer viable treatment alternatives in many cases. i.v.Ig and anti-D are both equally efficacious; however, anti-D therapy may come with lower cost to the patient and the healthcare system. Thus, in treating patients with ITP, clinical and economic considerations should be considered when choosing among the various treatment options. PMID- 11914000 TI - Alloimmune thrombocytopenia of the fetus and the newborn. AB - Fetal/neonatal alloimmune thrombocytopenia results from a maternal alloimmunization against fetal platelet antigens. Care must be taken in making a correct diagnosis that eliminates other causes of thrombocytopenia that may occur during pregnancy. Biological diagnosis is normally made by platelet genotyping and search for the maternal alloantibody using monoclonal antibodies in an antigen capture assay. Fetal alloimmune thrombocytopenia, when severe, may result in intracerebral hemorrhage leading to hydrocephalus and death of the fetus. Neonatal alloimmune thrombocytopenia may be present in an otherwise healthy infant. While screening procedures are not in place to detect fetal/neonatal alloimmune thrombocytopenia, it is true that fetal hydrocephalus, unexplained fetal thrombocytopenia with or without anemia, or recurrent miscarriages should be adequate indicators for suspecting fetomaternal alloimmune thrombocytopenia. Multiparous women with a history of giving birth to at least one alloimmune thrombocytopenic infant should be carefully monitored in subsequent pregnancies. Postnatal management of neonatal alloimmune thrombocytopenia involves compatible platelet transfusion in the neonate. Antenatal management of fetal alloimmune thrombocytopenia is controversial and includes a combination of maternal intravenous gamma globulin (i.v.IgG) administration, intrauterine platelet transfusions, and corticosteroid therapy, while monitoring fetal platelet counts closely throughout the course of pregnancy. Screening of pregnant women may become a public health issue only after antenatal therapy is more standardized. PMID- 11914001 TI - HIV-related thrombocytopenia. AB - Chronic thrombocytopenia is a common hematologic disorder in patients infected with the human immunodeficiency virus (HIV). Although often asymptomatic, the thrombocytopenia may be associated with a variety of bleeding abnormalities. The underlying pathophysiology includes accelerated peripheral platelet destruction and decreased ('ineffective') production of platelets from the infected megakaryocytes. HIV-related thrombocytopenia (HIV-TP) responds to antiretroviral therapy. Most studies have evaluated the use of AZT (zidovudine) and have shown increased platelet production. Combination therapy (HAART) also resulted in sustained platelet increases. When antiretroviral agents fail to improve the platelet count or cannot be used, other therapies, similar to those used in 'classic' immune thrombocytopenia (ITP), can be employed, including steroids, intravenous immunoglobulin (i.v.intravenous anti-D or splenectomy. Anti-D treatment offers advantages for HIV-TP because the duration of effect appears to be significantly longer than the response duration after i.v. therapy (initial results of our open-arm study were confirmed by our randomized trial). Of note, follow-up of heavily treated patients showed no acceleration of CD4 decline and no change in plasma viral load measurements. Splenectomy has been used to treat HIV-positive patients with refractory thrombocytopenia. Although it is effective therapy, there are concerns about infections and selection of appropriate candidates. Other treatment modalities, such as interferon, vincristine, danazol, low-dose splenic irradiation and staphylococcal protein A immunoadsorption have shown limited success in HIV-TP. Alternatively, thrombocytopenia in HIV-infected patients may be treated with pharmacological hyperstimulation of megakaryocytopoiesis (administration of PEG-rHuMGDF or TPO). Latest evidence indicates that the chemokine receptor CXCR4 (coreceptor for the cellular entry of lymphotropic HIV strains) is expressed on megakaryocytes; as a result, the development of chemokine receptor antagonists may modify the course of the disease. PMID- 11914002 TI - Heparin-induced thrombocytopenia: an overview. AB - Heparin-induced thrombocytopenia (HIT) is the most important immunological drug reaction that patients face today. HIT typically develops in patients 5 days after starting heparin therapy, but can occur sooner with recent heparin exposure or rarely have a delayed onset. The platelet count typically drops below 150 x 10(9)/L (average 60 x 10(9)/L), and patients may experience a thrombotic episode simultaneously or shortly after the onset of thrombocytopenia. The thrombocytopenia and the associated thrombotic episodes are now considered to be overlapping outcomes of the same syndrome. The pathophysiology of HIT has been characterized: immune complexes of IgG and heparin in association with a small platelet peptide, platelet factor 4 (PF4), activate platelets by binding to the Fc receptors (FcR) and releasing procoagulant-active, platelet-derived microparticles. The recognition that HIT is characterized by intense thrombin generation dictates the use of antithrombin agents in HIT therapy. Therapeutic approaches that are currently prevalent in the management of HIT will be discussed. PMID- 11914003 TI - Removal of knotted intravascular devices. Case report and review of the literature. AB - OBJECTIVE: to review published reports on knotted intravascular devices/catheters. METHOD: report of two cases and systematic review of the literature. RESULTS: a total of 113 reported cases of knotted intravascular devices/catheters were located. Pulmonary artery catheters (Swan-Ganz) were responsible for more than two thirds of the total reported intravascular knots. In 62% (70/113) of the cases withdrawal of the knotted catheters was achieved successfully with different interventional radiological techniques, avoiding the need for surgical exploration. In 32% (36/113) of the patients surgical removal was favoured. Capture with one of the interventional techniques and pulling down the knot into an easily accessible vein to be removed through an open venotomy, was the most common surgical procedure. However, in five cases, an open cardiotomy was required. In seven cases the patient's condition was critical and precluded any surgical procedure, so the knotted catheter was left in situ. The mortality of this event was 8% (9/113). CONCLUSIONS: interventional radiological techniques have largely replaced open surgical removal. Knotted catheters may need to be surgically removed when (a) the knot is large in size with many loops, or (b) intracardiac fixing of the knot is encountered. PMID- 11914004 TI - Eversion vs conventional carotid endarterectomy: a systematic review. AB - OBJECTIVE: to determine whether eversion carotid endarterectomy (CEA) was safe and more effective than conventional CEA. METHODS: controlled trials comparing eversion vs conventional technique for CEA were identified from the Cochrane Stroke Review Group database plus additional hand searching. Researchers were contacted to identify additional published and unpublished studies. Randomised and pseudorandomised trials comparing eversion to conventional techniques in patients undergoing CEA were examined. Outcomes included stroke and death, carotid restenosis/occlusion, and local complications. RESULTS: five trials were included comprising 2465 patients and 2590 arteries. There were no significant differences in the rate of perioperative stroke or death (1.7% vs 2.6%, odds ratio [OR] 0.44, 95% confidence interval [CI] 0.10-1.82) and stroke during follow up (1.4% vs 1.7%; OR: 0.84; 95% CI: 0.43-1.64) between eversion and conventional CEA techniques. Eversion CEA was associated with a significantly lower rate of restenosis >50% during follow-up (2.5% vs 5.2%, OR: 0.48, 95% CI: 0.32-0.72). There were no statistically significant differences in local complications between the eversion and conventional group. When eversion procedures were compared with patch procedures only, non-significant differences were found in primary outcomes. CONCLUSIONS: eversion CEA may be associated with low risk of arterial occlusion and restenosis. However, numbers are too small to definitively assess the benefits and disadvantages of eversion CEA. Reduced restenosis rates did not appear to be associated with clinical benefit in terms of reduced stroke risk, either perioperatively or later. Until further evidence is available, the choice of the CEA technique should be based on the experience and familiarity of the individual surgeon. PMID- 11914005 TI - Recent strategies to reduce vein graft occlusion: a need to limit the effect of vascular damage. AB - Despite early identification and aggressive modification of atherosclerotic risk factors, many patients still require surgical revascularisation for established atherosclerotic vascular disease. However, bypass surgery is hampered by a high incidence of vein graft failure. New strategies are being introduced to improve these results, with early data suggesting that improved patency rates are possible. These vary from the use of adjuvant pharmacological agents and local gene transfer strategies to the modification of vein harvesting techniques in order to reduce vascular damage to all layers of the graft. Advances in vascular biology have resulted in new insights into the role of the endothelium and adventitia in vein graft remodelling. Although recent pharmacological adjuvant therapy and molecular techniques have been described that may be used to reduce the incidence of vein graft occlusion a more desirable approach for improved graft patency rates may be achieved simply by using atraumatic surgical techniques aimed at minimising vascular damage during vessel harvesting and subsequent anastamoses during bypass surgery. PMID- 11914006 TI - POSSUM and P-POSSUM overpredict mortality for carotid endarterectomy. AB - OBJECTIVES: the aim was to test the predictive accuracy of POSSUM and P-POSSUM on patients undergoing CEA. DESIGN: retrospective and prospective study. MATERIALS: 499 CEAs performed by four vascular surgeons from a single unit from 1992-99. The median age was 68 (range 38-86) and 60% were men. METHODS: physiological parameters, operative parameters and the 30-day mortality were collected. Predicted mortality for each patient was calculated using POSSUM and P-POSSUM equations. Patients were stratified into risk groups and observed and predicted deaths were compared. Accuracy of the prediction was assessed using chi-squared analysis. RESULTS: the observed 30-day mortality was 1.8% (9/499). The predicted deaths using POSSUM and P-POSSUM analysis were 49 and 25 respectively compared to nine observed deaths. There was significant evidence of lack of fit of both models in predicting mortality (chi-squared analysis, p<0.05). CONCLUSIONS: POSSUM and P-POSSUM overpredict mortality and are unsuitable for comparative audit of CEA. Models developed specific for CEA might accurately predict mortality. PMID- 11914007 TI - Distal percutaneous transluminal angioplasty through infrainguinal bypass grafts. AB - AIM: to evaluate the results of transluminal angioplasty (PTA) performed through infrainguinal bypass grafts for stenotic or occlusive lesions at the distal anastomosis and/or in the runoff arteries. DESIGN: retrospective clinical study. MATERIAL AND METHODS: forty-one patients underwent 57 procedures at the distal anastomosis (n=13), in the runoff arteries (n=32) or at both locations (n=12) at a median of 9.6 months (range, 2-76 months) after infrainguinal bypass grafting. Nineteen procedures were on the popliteal artery, the rest on the crural arteries. Eleven procedures related to occlusions less than 5 cm in length. RESULTS: technical success was achieved in 91%. Primary and primary assisted graft patency rates at 3 years were 32% and 53%, respectively. There were no significant differences in patency rates with regard to the graft material, the type of lesion, the level of PTA, the status of runoff and the use of thrombolysis before PTA. No patients underwent amputation as a direct consequence of failed PTA or graft occlusion. One patient underwent acute surgical intervention due to graft occlusion at the time of attempted PTA. CONCLUSION: the results of PTA at the distal anastomosis and/or in the runoff arteries in limbs with infrainguinal bypass seemed to be inferior to the results of surgical revisions reported in literature. However, as failed PTA did not jeopardise vein patch angioplasty or jump grafting, it is a reasonable alternative to surgical intervention in selected cases. PMID- 11914008 TI - Platelet and leukocyte activation during aortoiliac angiography and angioplasty. AB - OBJECTIVE: to evaluate platelet and leukocyte activation during aortoiliac angiography and percutaneous transluminal angioplasty (PTA). METHODS: an observational study of 14 patients with aortoiliac atherosclerotic disease, nine of whom underwent PTA. The proportion of fibrinogen-, and P-selectin positive platelets, P-selectin expression on platelets, intraplatelet cGMP and cAMP, CD18 positive granulocytes, CD18 expression on granulocytes, plasma (p)-neopterin, p TNF alpha and p- interleukin-6 were repeatedly measured in arterial blood during angiography and in venous blood before and after. RESULTS: compared to a previous venous sample, arterial intraplatelet cAMP was increased proximal to the atherosclerotic lesion before contrast infusion and PTA (median 18 [range: 14-22] vs 16 [15-21] pmol/10(9) platelets p<0.05), and intraplatelet cGMP was increased proximal to the lesion after contrast infusion and PTA (1.2 [0.8-3.9] vs 0.9 [0.6 2.5] pmol/10(9) platelets p<0.05). Four hours after angiography, both the proportion of P-selectin positive platelets (28[11-55]%) and platelet P-selectin expression (9[6-40]) had decreased (p<0.05), from arterial values distal to the lesion before contrast infusion and PTA (57 [24-78]% and 26 [10-83]). Granulocyte CD18 expression was lower during angiography than in a previous venous sample. CONCLUSIONS: the results are compatible with platelet but not leukocyte activation during peripheral angiography. PMID- 11914010 TI - Pirfenidone inhibits early myointimal proliferation but has no effect on late lesion size in rats. AB - AIMS: intimal hyperplasia is mediated by smooth muscle cell proliferation, migration and deposition of extracellular matrix. The anti-fibrotic agent pirfenidone has been shown to inhibit pro-fibrotic growth factors in non-vascular inflammatory models. This study investigated the effect of the novel anti fibrotic agent pirfenidone on the development of neointima. METHODS: male Sprague Dawley rats received either standard diet or diet supplemented with pirfenidone (250, 500, 1000 mg/kg/day). Animals underwent left common carotid balloon angioplasty and were explanted at 4, 8, 14 and 28 days and analysed for intimal thickening, pro-fibrotic gene expression, extracellular matrix deposition and metalloproteinase activity. RESULTS: neointimal thickness was significantly reduced in a dose-dependent manner at 14 days; pirfenidone 250 mg/kg (p<0.005), pirfenidone 500 mg/kg (p<0.001), pirfenidone 1 g/kg ( p<0.001). There were no significant differences in intimal thickening at 28 days. Expression of MMP-2, MMP-9, TIMP-1, collagen III and TGF-beta were all significantly inhibited at 14 days. Both collagen III expression and ECM deposition were reduced at 28 days ( p<0.05 and <0.002 respectively). CONCLUSION: pirfenidone reduces expression of MMPs governing smooth muscle cell proliferation and migration (MMP-2 and 9), and genes favouring ECM accumulation (TIMP-1 and collagen III). This study shows that inhibition of MMP activity is not sufficient to inhibit late lesion size. PMID- 11914009 TI - Local delivery of NO-donor molsidomine post-PTA improves haemodynamics, wall mechanics and histomorphometry in atherosclerotic porcine SFA. AB - OBJECTIVES: we investigated the therapeutic effect of angioplasty with local drug delivery (LDD) of the wall-accumulating NO-donor molsidomine (M) in the superficial femoral arteries (SFA) of atherosclerotic swine. MATERIALS AND METHODS: atherosclerotic Pietrin swines (n=14) underwent PTA-LDD-M (4 mg/2 ml) vs contralateral PTA-LDD-Placebo in the SFA using a channelled balloon angioplasty catheter. Invasive and colour Doppler energy (CDE) assessments of haemodynamics and wall mechanics were performed at 24 h (n=4) and 5 months (n=10). Immuno histolabelling of cell proliferation and histomorphometry were serially performed in perfusion fixed SFA samples. RESULTS: at 24 h, PCNA-positive nuclei revealed 33+/-14 and 12+/-3 proliferating cells/mm2 at placebo and molsidomine PTA-LDD sites, respectively (p<0.001). At 5 months, PTA-LDD-M vessels, compared with PTA LDD-P, had increased compliance (66+/-9 vs 11+/-4 ml/mmHg) and lowered impedance (0.11+/-0.05 vs 0.45+/-0.14 mmHg/ml x min(-1)) (p<0.05). CDE revealed low, middle and high velocity peaks at 7.5, 20 and 35, and 8, 15 and 22 cm x s(-1) in systolic and diastolic flows, respectively; and PTA-LDD-M prevented emergence of restenosis-associated increases in low blood velocities (p<0.01). PTA-LDD-M inhibited restenotic intimal thickening and medial thinning which decreased mean lumenal diameter in placebo-treated (2.6+/-0.3) as compared to molsidomine treated (3.4+/-0.3 mm) vessels (p<0.05). CONCLUSIONS: in the atherosclerotic porcine SFA model, PTA-LDD with molsidomine consistently improved haemodynamic wall mechanics, lowered cell proliferation and prevented late lumen loss observed with PTA-LDD with placebo. PMID- 11914011 TI - Bacterial colonisation of Doppler probes on vascular surgical wards. AB - AIM: hospital acquired infections cost the NHS 1 pound sterling billion each year and medical equipment may act both as source and vector of nosocomial infection. This study examined bacterial contamination of Doppler ultrasound probes (USP) in routine use on vascular surgical wards in six hospitals and the knowledge of staff about the potential for cross infection from contaminated probes. METHODS: probe head impressions and swab cultures of probe holders were plated on mannitol salt agar before and after cleaning with a paper towel. Putative S. aureus isolates were identified to species level and susceptibility to selected antimicrobials tested. Concurrently, junior medical staff were surveyed about probe cleaning protocols. RESULTS: methicillin susceptible S. aureus was isolated from 2/21 (10%) with near confluent bacterial growth from six others (28%). The latter may have obscured low numbers of S. aureus. Further since swabs were plated without prior enrichment culture, it is likely that contamination with S. aureus might have been underestimated. No positive cultures were obtained after wiping the USP with a paper towel. 22/23 (95%) junior doctors failed to clean the USP prior to use. CONCLUSION: USP contamination with pathogenic bacteria occurs under "in-use" conditions and junior medical staff are unaware of simple measures to prevent this. Strict guidelines for USP cleaning between patient use should, therefore, be adopted particularly when monitoring postoperative graft patency. PMID- 11914012 TI - Chronic aortic dissection not a risk factor for neurologic deficit in thoracoabdominal aortic aneurysm repair. AB - OBJECTIVE: chronic aortic dissection has long been considered a risk factor for neurologic deficit following thoracoabdominal aortic aneurysm (TAA) surgery. We reviewed our experience with regard to aneurysm extent and the use of adjunct, (distal aortic perfusion/cerebrospinal fluid drainage), and examined the impact of these factors on neurologic deficit among chronic dissection and non dissection cases. METHODS: between February 1991 and March 2001, we repaired 800 aneurysms of the descending thoracic and thoracoabdominal aorta. Seven hundred and twenty-nine cases were elective; 196 chronic dissection, 533 non-dissection. 182/729 (24.9%) were TAA extent II. Among these, 61/182 (33%) involved chronic dissection. Adjunct was used in 507/729 (69.6%). We conducted detailed multivariate analyses to isolate the impact of chronic aortic dissection on neurologic morbidity, with other important risk factors taken into account. RESULTS: overall, 32/729 (4.4%) patients had neurologic deficit upon awakening; 7/196 (3.6%) in chronic dissections, and 25/533 (4.7%) in non-dissections. Adjunct had a major effect, reducing neurologic deficit in TAA extent II from 10/36 (27.8%) to 10/146 (6.9%) (p=0.001). However, in univariate and multivariate analysis, chronic dissection did not increase the risk of neurologic deficit, regardless of extent or mode of treatment. CONCLUSION: in contrast to previous reports, we determined that chronic aortic dissection is not a risk factor in TAA patients. PMID- 11914013 TI - Acute leg ischaemia from thrombosed popliteal artery aneurysms: role of preoperative thrombolysis. AB - OBJECTIVE: to evaluate early and long term results of thrombolysis and surgery in acutely thrombosed popliteal artery aneurysms. SETTING: retrospective study; teaching hospital. MATERIALS: between 1990 and 2000, 109 popliteal artery aneurysms were operated on. In 24 patients acute leg ischaemia due to thrombosis of aneurysm was present. METHODS: ten patients underwent urgent surgical intervention (group 1); and 14 thrombolysis with urokinase, until patency of popliteal and tibial vessels was achieved or for a maximum of 3 days (group 2). Ultrasonographic follow-up was performed at 1, 3, 6 and 12 months and then annually. Early results and follow-up data were analysed by chi-square test and life-table analysis (Kaplan-Meier curve) and late results were compared by mean of log rank test. RESULTS: in group 1 early limb salvage was 70%; in group 2 it was 86% (p=n.s.). When thrombolysis was successful, patency and limb salvage were 100%. There was no local or systemic complications during thrombolysis nor worsening of ischaemia. Follow-up was completed in 91 cases, with a mean duration of 26 months. Forty-eight months primary patency rate was better, even if not statistically significant, in group 2 than in group 1. CONCLUSIONS: in patients with acute leg ischaemia due to thrombosis of popliteal artery aneurysms, preoperative thrombolysis can be considered a safe and effective alternative to urgent surgery. PMID- 11914014 TI - Delayed presentation of traumatic popliteal artery pseudoaneurysms: a review of seven cases. AB - OBJECTIVES: to examine the management and outcome of patients with traumatic popliteal artery pseudoaneurysms with delayed presentation. MATERIALS: over a 2 year period 7 patients with traumatic popliteal pseudoaneurysms presenting to a tertiary referral unit after a delay of 1 month were reviewed. METHODS: a retrospective review of clinical records. RESULTS: the median interval between injury and presentation was 1.5 months (range 1-24 months). Penetrating trauma from gunshot wounds was the cause of the initial vascular injury in 6 patients and a stab wound in one. All patients had large pseudoaneurysms of more than 8 cm filling the popliteal fossa with variable degrees of fixed flexion deformity (FFD) of the knee. Severe FFD of more than 40 degrees was noted in 2 patients and 3 others had mild flexion deformities of 10 to 20 degrees. Six patients underwent repair of the pseudoaneurysm. One patient required an above knee amputation due to an infected false aneurysm. There were no deaths or graft related complications. Despite aggressive post-operative physiotherapy only 2 patients were able to completely straighten the leg at the time of discharge. CONCLUSIONS: in patients with neglected popliteal artery pseudoaneurysms, morbidity is associated with the pre-operative degree of fixed flexion deformity of the knee. PMID- 11914015 TI - Up-regulation of MMP-2 and MMP-9 leads to degradation of type IV collagen during skeletal muscle reperfusion injury; protection by the MMP inhibitor, doxycycline. AB - OBJECTIVES: to determine the role of matrix metalloproteinases, MMP-2 and MMP-9, in reperfusion injury following skeletal muscle ischaemia and whether inhibition of MMPs by doxycycline protects against tissue damage. METHODS: rats were anaesthetised and a tourniquet applied to the proximal thigh to occlude blood flow. Four hours of ischaemia was followed by reperfusion for 0, 4, 24 or 72 h. Two further groups received doxycycline for 7 days prior to bilateral ischaemia and 24 h reperfusion. Skeletal muscle from both limbs, kidneys and lungs were harvested for zymography and immunohistochemical staining for type IV collagen. RESULTS: upregulation of MMP-2 and MMP-9 was detected by zymography in the ischaemic leg and lung but not in the kidney. Quantitative immunohistochemical analysis showed marked degradation of type IV collagen in reperfused muscle, lung and kidney. Doxycycline-treated rats showed significant preservation of type IV collagen in skeletal muscle and a trend towards preservation in kidney and lung. CONCLUSIONS: MMP-2 and MMP-9 are strongly upregulated in skeletal muscle ischaemia/reperfusion injury and are also upregulated in remote organs, leading to degradation of basement membranes. Inhibition of MMP activity may therefore be potentially therapeutically useful in reducing the severity of reperfusion injury. PMID- 11914016 TI - Expectations for the skills of final year medical students in examining lower limb arteries and veins. PMID- 11914017 TI - A rare case of mycotic aortic pseudoaneurysm. PMID- 11914018 TI - Retrograde subintimal angioplasty via a popliteal artery approach. PMID- 11914019 TI - Reflections from a 40 year journey with growth hormone and IGF-I. PMID- 11914020 TI - Clinical implications of the IGF-cancer connection. PMID- 11914021 TI - Are the insulin-like growth factors relevant to cancer? AB - Recognition of molecular pathways relevant to cancer biology have led to advances in prevention and treatment. Numerous laboratory and clinical investigations have implicated the insulin-like growth factors (IGFs) in tumourigenesis. In this review, the evidence for the involvement of IGFs in cancer is discussed. While these data are persuasive, it is clear that additional methods to regulate IGF action in cancer patients are needed to substantiate the role of this growth factor family in cancer biology. PMID- 11914022 TI - Roles of insulin-like growth factor-I (IGF-I) and IGF-I binding protein-2 (IGFBP2) and -5 (IGFBP5) in developing chick limbs. AB - Insulin-like growth factor-I (IGF-I) and the IGF-I binding proteins (IGFBPs) which modulate IGF-I action have been implicated in the development of the vertebrate limbs and skeleton. We have examined the distribution of IGF-I, IGFBP2 and IGFBP5 in developing chick limb buds and have investigated their functional roles and relationships during chick limb development. IGF-I and IGFBP2 are co expressed throughout the lateral plate from which limbs form, although IGFBP2, unlike IGF-I, does not promote formation of rudimentary limb buds from non-limb forming flank regions in vitro. During limb outgrowth, IGF-I is present in non AER limb ectoderm, but little IGF-I is present in the AER itself, suggesting that restriction of endogenous IGF-I activity may be required for proper AER function. Consistent with this possibility, the ectoderm of mutant limbless and wingless wing buds, which fail to form an AER, continues to express IGF-I. We also found that the AER contains abundant IGFBP2 but that IGFBP2 is not present in limb subridge mesoderm. In contrast, IGFBP2 is present in the distal mesoderm of mutant limbless or wingless limb buds, which fail to grow out. This suggests that attenuation of IGFBP2 expression is controlled by the AER and that cessation of IGFBP2 expression may be necessary for the proliferation and suppression of differentiation of subridge mesoderm that is required for limb outgrowth to occur. Consistent with this possibility, we found that exogenous IGFBP2 inhibits the anti-differentiative activity of the AER in vitro. We also found that regions of cell death in the limb contain abundant IGF-I-immunoreactive cells, consistent with a role for IGF-I in apoptosis. During skeletogenesis, IGF-I and IGFBP2 are co-localized to the condensing central core of the limb, implicating these factors as potential regulators of the onset of chondrogenic differentiation. Intriguingly, we found that IGF-I and IGFBP2 have opposing effects on chondrogenesis, as IGF-I stimulates but IGFBP2 inhibits accumulation of cartilage matrix by micromass cultures in vitro. Long [R(3)] IGF-I, an analog of IGF-I that cannot bind IGFBPs, is more effective than IGF-I in stimulating matrix accumulation, consistent with a negative role for IGFBP2 in chondrogenesis. As the chondrocytes of the limb mature, IGF-I is present only in terminal hypertrophic chondrocytes, which undergo programmed cell death, while IGFBP2 becomes localized to prehypertrophic and hypertrophic chondrocytes, suggesting involvement in chondrocyte maturation. Consistent with this possibility, we found that exogenous IGFBP2 induces precocious expression of Indian hedgehog, a marker of prehypertrophy, in maturing chondrocytes in vitro. IGF-I and IGFBP2 are also present in the osteoblasts, clasts and nascent matrix of the long bones, consistent with roles in endochondral bone formation. Unlike in rodent limbs, IGFBP5 is not expressed by chick limb ectoderm or AER. IGFBP5 expression is highly localized to developing limb musculature and, later, to the developing skeletal elements where it is expressed by osteoblast precursers and osteoblasts. The results of this study suggest potential novel roles for IGF-I and IGFBP2 in several aspects of limb development including limb outgrowth and AER activity, programmed cell death, chondrogenesis and chondrocyte maturation. PMID- 11914023 TI - Reduced serum insulin-like growth factor-1 (IGF-1) and IGF-binding protein-3 levels in adults with inflammatory bowel disease. AB - In the present study, the changes in circulating IGF-1 and its binding protein IGFBP-3 were determined in adult patients with active inflammatory bowel disease (IBD) in order to assess the effect of this inflammatory condition on the IGF system. IGF-1 and IGFBP-3, as well as interleukin-6 (IL-6) were measured in serum obtained from 22 consecutive newly diagnosed patients (mean age 41.3 years) with active IBD, including 10 patients with Crohn's disease (CD), and 12 with ulcerative colitis (UC). For comparison the same parameters were determined in 30 healthy volunteers matched for age, sex and Body Mass Index (BMI). Serum IGF-1 and IGFBP-3 levels were similar in the two subgroups of patients and the values from all patients were combined for comparison with those from the control group. The mean (+/- SD) serum IGF-1 concentration (178 +/- 91 ng/ml) in the patients with IBD was lower compared with that in the controls (227 +/- 79 ng/ml, P<0.035). Similarly, the mean IGFBP-3 concentration in the patients was lower than in the controls (1.6 +/- 0.6 ng/ml vs 3.2 +/- 0.7 ng/ml respectively, P<0.001), Serum IL-6 levels were higher in the patients compared with the controls (5.5 +/- 4.2 vs 0.65 +/- 0.11 pg/ml, P<0.0001). The reduced IGF-1 and IGFBP-3 levels in patients with active IBD suggest that this systemic inflammatory condition is associated with a degree of acquired GH resistance, possibly induced by inflammatory cytokines. PMID- 11914024 TI - Inhibition of human osteoblast marker gene expression by retinoids is mediated in part by insulin-like growth factor binding protein-6. AB - All-trans -retinoic acid (atRA) inhibits osteoblast marker gene expression and markedly increases expression of insulin-like growth factor binding protein-6 (IGFBP-6) in human osteoblasts. The possibility that IGFBP-6 inhibits the osteoblast phenotype and also mediates the inhibitory effect of atRA on osteoblast marker gene expression was explored using an antisense approach. Stable human osteoblast-like osteosarcoma SaOS-2 cells were prepared that expressed antisense IGFBP-6 RNA under basal and atRA-stimulated conditions. The functional expression of IGFBP-6 antisense RNA was confirmed by measuring IGFBP-6 mRNA by Northern analysis or by measuring IGFBP-6 protein in the conditioned media (CM) by radioimmunoassay. Antisense clones produced less mRNA and had less IGFBP-6 protein in the CM than controls. IGFBP-6 protein levels in the CM were inversely correlated with alkaline phosphatase (ALP) activity, whereas IGFBP-3 and IGFBP-4 protein levels were not. We reasoned that atRA would have little or no effect on ALP activity in IGFBP-6 antisense clones if atRA mediated its inhibitory effects by recruiting IGFBP-6. In the majority of IGFBP-6 antisense clones with the lowest IGFBP-6 mRNA and CM protein levels and only modest changes in other IGF system components, atRA did not significantly decrease ALP activity. These findings provide evidence that atRA recruits IGFBP-6 to inhibit the human osteoblast phenotype. PMID- 11914025 TI - A novel and de novo splice-donor site mutation in intron 3 of the GH-1 gene in a patient with isolated growth hormone deficiency. AB - Heterozygous mutations at the splice-donor site of inron 3 of the GH-1 gene are known to affect growth hormone (GH) mRNA splicing and cause isolated GH deficiency (IGHD), which is inherited in an autosomal dominant trait. We report here a novel and de novo heterozygous IVS3 + 6T --> G mutation of the GH-1 gene in a Japanese patient with IGHD. RT-PCR analyses of the GH-1 minigene transcripts demonstrated that the IVS3 + 6T --> G mutation causes complete skipping of exon 3. We found a heterozygous IVS4 + 18G --> T polymorphism of the GH-1 gene in the patient, which was shared by the father, but not by the mother. Sequencing of individual alleles of the patient's GH-1 gene confirmed that the IVS3 + 6T --> G mutation and the IVS4 + 18G --> T polymorphism exist on the same chromosome. These findings indicate that the IVS3 + 6T --> G mutation arose in a germ cell of the father and caused IGHD in the patient. PMID- 11914026 TI - Total and free insulin-like growth factor I, insulin-like growth factor binding protein 3 and acid-labile subunit reflect clinical activity in acromegaly. AB - The aim was to evaluate, markers of disease activity in acromegaly in relation to perceived disease activity. Thirty-seven consecutively treated, acromegalic patients, classified by clinical symptoms as inactive (n=16), slightly active (n=10) and active (n=11), entered the study. When evaluating the inactive and the active groups, we found that positive and negative predictive values (PV(pos), PV(neg)) for clinical disease activity of total and free insulin-like growth factor-I (IGF-I) were 0.59, 0.90 and 1.00, 0.82 respectively. Acid-labile subunit (ALS) showed diagnostic merit similar to insulin-like growth factor binding protein-3 (IGFBP-3) with PV(pos) of 0.69 and 0.71 and PV(neg) of 0.91 and 0.92 respectively. We conclude that free IGF-I is more closely related than total IGF I to perceived disease activity and is as such useful when evaluating previously treated acromegaly for disease activity. Total IGF-I, IGFBP-3 and ALS possess a higher PV(neg) for the clinical disease activity. None of the parameters can at present be claimed to be superior to the others and thus all the measured parameters are recommended to be part of the evaluation of acromegalic patients. PMID- 11914027 TI - Maternal nutrition affects the ability of treatment with IGF-I and IGF-II to increase growth of the placenta and fetus, in guinea pigs. AB - The aim of this study was to investigate how administration of IGF-I and IGF-II, during early to mid pregnancy, affects maternal growth and body composition as well as fetal and placental growth, in ad libitum fed, and in moderately, chronically food restricted guinea pigs. From day 20 of gestation, mothers (3-4 months old) were infused with IGF-I, IGF-II (565 microg/day) or vehicle for 17 days and then killed on day 40 of gestation. Maternal organ weights, fetal and placental weights were assessed. Treatment with IGFs did not alter body weight gain and had small effects on body composition in the mothers. Both IGF-I and IGF II increased fetal and placental weights in ad libitum fed dams and IGF-I increased placental weight in food restricted dams. In conclusion, treatment with IGF-I during the first half of pregnancy stimulates placental growth in both ad libitum fed and food restricted guinea pigs without affecting maternal growth while fetal growth is stimulated by IGF treatment only in ad libitum fed animals. PMID- 11914028 TI - Depolarization induces insulin-like growth factor binding protein-2 expression in vivo via NMDA receptor stimulation. AB - The effect of depolarization and N-methyl-D-aspartate (NMDA) receptor blockade on insulin-like growth factor-I (IGF-I), IGF binding protein-2 (IGFBP-2) and IGFBP-4 expression was analysed in vivo. Depolarization was induced in adult rat brains by applying 3 M KCl to the exposed cortex for 10 min. A subgroup of animals also received daily injections of MK-801. Four days after KCl exposure, the brains were analysed by in situ hybridization, immunohistochemistry and TUNEL. A significant upregulation of IGFBP-2 mRNA and protein was detected in astrocytes after KCl exposure This upregulation was reduced by MK-801 treatment. No alterations in IGF-I or IGFBP-4 mRNA levels were noted. We did not detect TUNEL positive cells, morphological signs of necrosis or apoptosis, or neuronal loss in the depolarized zone. Taken together, these findings indicate that upregulation of IGFBP-2 by depolarization is mediated by NMDA receptors, and, as no neuronal damage was detected, astrocytic NMDA receptors may be responsible for this upregulation. PMID- 11914029 TI - The changes of IGF binding proteins after rhGH administration to patients totally dependent on parenteral nutrition. AB - The objective was to study the effect of recombinant human growth hormone (rhGH) administration to patients with chronic malnutrition maintained on total parenteral nutrition (TPN) on the levels of insulin-like growth factor-I (IGF-I) and IGF binding proteins (IGFBPs) during a double-blind trial. After 1 week of TPN the patients were randomized into group I (placebo) or group II (rhGH). Samples were collected on the first day (start of the TPN) to measure basal values, the seventh day to study the effect of TPN and the 10th, 14th and 21st days to evaluate the rhGH effect. Basal laboratory evaluation, nutritional status and glucose tolerance were assessed using standard laboratory techniques. Radioimmunoassays were used to analyse IGF-I, free IGF-I (fIGF-I) and IGFBP1-3. Electrophoresis with Western ligand blotting and Western immunoblotting was applied to find the pattern of IGFBPs. TPN had no effect on the circulating IGF-I concentration and the pattern of IGFBPs present in the studied groups of patients. The rhGH administration led to significant increases of IGF-I, total IGFBP-3, glycosylated IGFBP-3 (39, 42 kDa) and the 29 kDa fragment of IGFBP-3 and the decrease of IGFBP-2 during the trial (P<0.05). The mean levels of IGFBP-1, fIGF-I and the parameters of nutritional status in group II during the trial were not significantly influenced by rhGH. However, it has been found that IGFBP-1 and fIGF-I levels were correlated with the levels of the weekly nitrogen balance of each patient in group II at the end of the trial. In spite of the significant changes of IGF-I, IGFBP-2, total IGFBP-3 and IGFBP-3 (29 kDa proteolytic fragment) after rhGH administration to patients with malnutrition, maintained on parenteral nutrition, the increase of nitrogen balance was seen only in patients who decreased their IGFBP-1 and increased bioavailable IGF-I as reflected by measurement of fIGF-I. The levels of IGFBP-1 may provide a useful marker of IGF-I bioavailability in monitoring the efficiency of the rhGH therapy in malnourished patients. PMID- 11914030 TI - Mechanism of ruminant placental lactogen action: molecular and in vivo studies. AB - Ruminant placental lactogens (PLs) are structurally related to prolactins (PRLs) and growth hormones (GHs) and are secreted by placentae. Ruminant PLs are unusual in their capacity to bind and activate PRL and GH receptors (Rs) from other species. The present minireview summarizes several works showing that unlike in heterologous species (rat, rabbit, human), in homologous (ruminant) species, PLs act by activating PRLRs or by heterodimerizing GHRs and PRLRs, and suggests that this may be the main mechanism of PL action in vivo. Mutations impairing the ability of ovine (o)PL or bovine (b)PL to form complexes with PRLRs (but not with GHRs) do not cause loss of biological activity, because the transient existence of the homodimeric complex is still sufficient to initiate the signal transduction; however, mutants do lose their ability to activate homologous PRLRs. To explain this difference, we proposed a novel term-minimal time of homodimer persistence-which assumes that to initiate the signal transduction, a "minimal time" of homodimer existence is required for transphosphorylation of associated JAK2s. In interactions between ruminant PLs and homologous PRLRs, this minimal time is met through the interaction with homologous PRLRs, which has a shorter half-life than with heterologous PRLRs. Thus oPL or bPL are active in cells possessing both homologous and heterologous PRLRs. Mutations of PLs decrease the affinity, shortening the "time of homodimer persistence." In heterologous interactions, the minimal time is still sufficient to initiate the biological activity, whereas in homologous interactions, which in any case are weaker, further destabilization of the complex shortens its persistence below the minimal time, causing loss of biological activity. PMID- 11914031 TI - Somatic mutations in thyroid nodular disease. AB - Thyroid nodules can be found in up to 50% of inhabitants of iodine-deficient areas and are classified as hot or cold thyroid nodules according to their scintigraphic characteristics. Studies of hot thyroid nodules with comparable mutation detection methods and screening at least exon 10 of the TSH receptor reported frequencies for somatic TSH-receptor mutations ranging from 20 to 82% in patients with similar iodine supply. We have recently screened 75 hot thyroid nodules for somatic TSH-receptor mutations with the more sensitive DGGE method and found somatic TSH-receptor mutations in 57% and Gsalpha mutations in 3%. As 50% of the mutation-negative nodules from female patients are of monoclonal origin when tested for X-chromosome inactivation somatic mutations in other genes are likely to cause the development of hot thyroid nodules. Scintigraphically nonsuppressible areas have been identified in up to 40% of euthyroid goiters in iodine-deficient areas. We recently identified somatic TSH-receptor mutations in microscopic autonomous areas with increased 125T uptake in euthyroid goiters studied by autoradiography 20 years ago. These constitutively activating somatic TSH-receptor mutations in minute autoradiographically hot areas of euthyroid goiters are very likely starting foci which most likely lead to toxic thyroid nodules in iodine-deficient goiters. Therefore iodine deficiency does not only lead to euthyroid goiters but also to thyroid autonomy. The latter is also suggested by epidemiologic studies. Similar mechanisms induced by iodine deficiency and the subsequent hyperplasia, mutagenesis, and selection of cell clones could also lead to cold thyroid nodules by somatic mutations that only initiate growth but not hyperfunction of the affected thyroid epithelial cell. Somatic ras mutations have frequently been detected in histologically characterized thyroid adenomas or adenomatous nodules. However, they seem to be rare in cold thyroid nodules. Since the majority of these latter nodules and 60% of the cold thyroid nodules are monoclonal other somatic mutations are likely in these nodules. PMID- 11914032 TI - Cloning and characterization of human agmatinase. AB - Arginine decarboxylase (ADC) and agmatinase are part of an operon in Escherichia coli, which constitutes the primary pathway of polyamine synthesis from arginine. This pathway is also known to exist in plants, but until recently, neither agmatine nor ADC, the enzyme that synthesizes it, nor agmatinase the enzyme that is responsible for conversion of agmatine to putrescine, were known to exist in man or other mammals. We describe here the cloning of the agmatinase gene and the tissue distribution of its transcription product. Human agmatinase contains 352 amino acid residues and has a calculated molecular weight of 37,688 kDa. It has 56% similarity to E. coli agmatinase and 42% similarity to human arginases I and II and shares highly conserved substrate-binding domains with these well characterized enzymes. PMID- 11914034 TI - Identification and characterization of temperature-sensitive mild mutations in three Japanese patients with nonsevere forms of very-long-chain acyl-CoA dehydrogenase deficiency. AB - Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is clinically classified into severe, intermediate, and myopathic forms. We identified mutations in three unrelated Japanese patients with VLCAD deficiency: two with the myopathic form and one with the intermediate form, all compound heterozygotes of K264E/M437V, A416T/1798delA, and P89S/IVS16-3delAA, respectively. We characterized four missense mutations, K264E, M437V, A416T, and P89S, by transisent expression analysis, using SV40-transformed fibroblasts derived from a VLCAD-null patient, as recipient cells. In transient expression of the wild-type VLCAD cDNA, VLCAD activity at 30 degrees C was higher than at 37 degrees C. Moreover, this temperature-sensitive character is more evident in all the mutant proteins tested than in wild type. Based on characterization of the five missense mutations identified in four Japanese patients, including data on one patient with the myopathic form previously reported, patients with the nonsevere forms (intermediate or myopathic forms) have missense mutations with residual activities in at least one allele. Expression analysis at 30 degrees C may be more useful for evaluating these missense mutations, compared with that at 37 degrees C. PMID- 11914033 TI - Evaluation of liver fatty acid oxidation in the leptin-deficient obese mouse. AB - We hypothesized that liver fatty acid oxidation (FAO) is compromised in the leptin-deficient obese (Lep(ob)/Lep(ob)) mouse model, and that this would be further challenged when these mice were fed a high-fat diet. Obese mice had a 3.8 fold increased body fat content and a 9-fold increased liver fat content as compared to control mice when both groups were fed a low-fat diet. The expression of liver FAO enzymes, carnitine palmitoyltransferase-1a, long-chain acyl-CoA dehydrogenase, medium-chain acyl-CoA dehydrogenase, and short-chain acyl-CoA dehydrogenase, was not affected in obese mice as compared to controls on either a low-fat or a high-fat diet. The expression of very-long-chain acyl-CoA dehydrogenase was elevated in obese mice on the control diet, as compared to control mice. For all measures evaluated, increasing the level of fat in the diet had a smaller effect than leptin deficiency. In summary, despite obese mice having an excess of fat available for mitochondrial beta-oxidation in liver, overall energy balance appeared to dictate that the net liver FAO remained at control levels. PMID- 11914035 TI - Characterization of six mutations in five Spanish patients with mitochondrial acetoacetyl-CoA thiolase deficiency: effects of amino acid substitutions on tertiary structure. AB - Mitochondrial acetoacetyl-CoA thiolase (T2) deficiency is an inborn error of ketone body and isoleucine metabolism. We identified and characterized 6 mutations, DelE85, K124R, A127V, Q145E, G152A, and E345V in 5 Spanish T2 deficient patients. Transient expression of mutant cDNAs was done at 37 and at 30 degrees C. Expression of the Q145E mutant cDNA resulted in about 12.5% normal amount at 37 degrees C and it retained 15% residual T2, indicating that specific activity of Q145E mutant protein was almost normal. This mutation reduced the heat stability of T2 activity. Although no significant residual activity was detected in either the G152A and A127V substitution, mutant proteins were detected, at 12.5% the normal amount at 37 degrees C and one-half normal at 30 degrees C for A127V, and 25 % only at 30 degrees C for G152A. Mutant proteins with Q145E, G152A, or A127V accumulated at 30 degrees C expression were stable for 48 h at 37 degrees C after cycloheximide treatment. Expression of DelE85, K124R, and E345V cDNAs gave neither residual T2 protein nor T2 activity. We constructed an improved tertiary structural model of T2 based on the X-ray crystal structure of acetoacetyl-CoA thiolase of Zoogloea ramigera. On the basis of this model, K124, A127, and G152 are located near the active site, mutations of which might affect catalytic function whereas Q145E, De185E, and E345V are distant from the active site with mutants being expected to destabilize the tertiary structure, especially during protein folding and dimerization. PMID- 11914036 TI - Developmental stage-specific expression of the alpha and beta subunits of the HIF 1 protein in the mouse and human fetus. AB - Erythropoietin (Epo) is a glycoprotein hormone that is the primary regulator of erythropoiesis. Transcription of the Epo gene increases in response to hypoxia or anemia. Epo is synthesized in the liver in fetal life and in the kidney later in gestation. In the mammalian fetus the switch in Epo production from the liver to the kidney occurs in the third trimester. Hypoxia-inducible factor (HIF-1) is a heterodimeric transcription factor consisting of an alpha and beta subunit that binds under hypoxic conditions to an enhancer element in the 3' region of the Epo gene. In order to determine if there is a relationship between expression of HIF 1 alpha and beta subunits with the shift in expression of the Epo gene from the liver to the kidney or with the transitional events occurring at birth we analyzed the expression of these mRNAs in mouse and human fetuses at different stages of gestation. Total RNA was extracted from the brain, heart, kidney, liver, and lungs of mice at P15, P17, and P19 of gestation, from newborn mice at Days 1 and 3, from an adult and an anemic adult mouse as well as from human fetuses at 14-22 weeks of gestation. RNA was analyzed by Northern blot and slot blot hybridization using appropriate cDNA probes. HIF-1alpha and -beta mRNA were expressed in all tissues tested and at all stages of gestation in the mouse and human fetus. Expression of HIF-1alpha and -beta in the mouse fetus was highest in the brain followed by heart, kidney, lung, and liver. Expression in the fetal and newborn mice was higher versus the adult and expression was higher in the anemic versus the normal adult mouse. In the human fetus a higher expression of HIF 1alpha was noted in the brain followed by heart, kidney, lung, and liver. There was a small trend toward a decrease in expression with advancing gestational age. HIF-1beta was expressed to a similar extent in all human tissues examined. Our studies indicate that expression of HIF-1alpha and -beta subunits was not related to the switch in Epo gene expression from the liver to the kidney. Although expression of HIF-1alpha and -beta did not decrease immediately after birth, it is possible that the HIF-1 protein is involved in the various events that occur during transition after birth. PMID- 11914037 TI - Modular design of a novel chimeric protein with combined thrombin inhibitory activity and plasminogen-activating potential. AB - In order to design plasminogen activators with improved thrombolytic properties we sought to construct the bifunctional protein HLS-2 which combines both a plasminogen-activating and an anticoagulative activity. The chimeric protein comprises four elements: a derivative of thrombin inhibitor hirudin, a 6-amino acid spacer, the sequence of plasminogen-activator staphylokinase (Sak), and a 13 amino acid expression tag at the C-terminus. The gene of the fusion protein was obtained by SOE-PCR, cloned into pCANTAB5E, and expressed in E. coli BL21. HLS-2 was purified from periplasmatic extracts and characterized by Western blotting. Plasminogen-activation of HLS-2 and of Sak in equimolar mixtures with plasminogen showed near equivalence as measured by plasmin-mediated cleavage of chromogenic substrate S-2403. For catalytic amounts of plasminogen-activator, however, HLS-2 was less effective by a factor of 1.7. HLS-2 also inhibited both the amidolytic and the fibrinolytic activities of thrombin. Similar concentrations of either commercial HV1 (42 pmol/L) or HLS-2 (250 pmol/L) were required to halve the initial rate of thrombin reaction with fluorogenic substrate Tos-Gly-Pro-Arg-AMC, suggesting the retention of high-affinity inhibition of thrombin by the fusion protein sufficiently strong to substitute anticoagulative comedication during fibrinolytic treatment. The results provide a rationale for further testing the efficacy of HLS-2 for the lysis of platelet-rich arterial blood clots and for the prevention of reocclusion after thrombolysis. PMID- 11914038 TI - Association of leucine 7 to proline 7 polymorphism in the preproneuropeptide Y with serum lipids in patients with coronary heart disease. AB - Leucine 7 (Leu7) to proline 7 (Pro7) substitution in the neuropeptide Y (NPY) gene has been associated with higher serum total and low-density lipoprotein (LDL) cholesterol levels, particularly in obese subjects. We investigated the frequency of the Pro7 allele and the association of the polymorphism with serum lipid levels in patients with coronary heart disease (CHD). A total of 414 CHD patients (mean age 61 years, range 33-74) participated in the cross-sectional EUROASPIRE study. Of the subjects 39% used lipid-lowering drugs. The frequency of the Pro7 allele in CHD patients (0.082) did not differ from that in control subjects (0.071). The mean (+/-SD) serum total cholesterol concentration was higher in women with the Pro7 allele (7.57 +/- 0.57 mmol/L, n = 8) than in women with the Leu7Leu genotype (6.69 +/- 1.01 mmol/L, n = 69, P = 0.019), when subjects using lipid-lowering medication were excluded. In contrast, serum total cholesterol concentration did not significantly differ between the genotypes in men. The Leu7Pro polymorphism was not associated with serum LDL, high-density lipoprotein (HDL) cholesterol, and triglyceride concentrations. In conclusion, the Pro7 allele in the NPY gene was associated with higher serum total cholesterol concentration only in women with CHD who did not use lipid-lowering drugs. PMID- 11914039 TI - Regulation of cell proliferation by insulin-like growth factor 1 in hyperoxia exposed neonatal rat lung. AB - Hyperoxic exposure of the developing lung leads to characteristic peribronchial and mesenchymal fibroproliferative changes. We hypothesize that O2-induced changes in the neonatal lung are mediated by Insulin-like growth factor 1 (IGF-1) and IGF-1 receptor (IGF-1R). Lung explant cultures were prepared from 3-day-old neonatal rat pups and exposed to room air or 95% O2 for 72 h. Western blots and immunohistochemistry were used to determine if hyperoxia stimulated IGF-1 and IGF 1R, and to identify the cell types involved. Retinoic acid was used to learn if this would inhibit oxygen-induced cell proliferation. Hyperoxia induced a significant increase in thymidine incorporation (control, 54 +/- 9; hyperoxia, 254 +/- 24 dpm/nM DNA; mean +/- SEM; N = 3; P < 0.05). This was inhibited by 5 x 10(-5) M RA (149 +/- 18 dpm/nM DNA; P < 0.05) and by anti-IGF-1 antibody (115 +/- 25 dpm/nM DNA; P < 0.05; N = 3). BrdU labeling in the mesenchymal cells was significantly increased in mesenchymal cells after exposure to oxygen (91% higher than the room air control) but not in epithelial cells. This increase was inhibited in the presence of retinoic acid. Western blots showed IGF-1 protein was increased after 72 h of O2 exposure compared to room air exposure (57 +/- 7 compared to 32 +/- 5 densitometric units; P < 0.05; N = 3). The increase was inhibited when the cultures were exposed to 95% O2 in the presence of anti-IGF-1 antibody (28 +/- 4; P < 0.05; N = 3). IGF-1 protein decreased in the presence of retinoic acid after oxygen exposure but not in room air. Immunostaining of O2 exposed lung showed IGF-1 was most abundant in airway and alveolar epithelial cells. We conclude that hyperoxia increases cell proliferation by stimulating IGF 1 in the neonatal rat lung. Interaction of IGF-1 and IGF-1R is an important cell cell communication mechanism in the developmental and repair processes of hyperoxic neonatal lung injury. PMID- 11914040 TI - Transfection screening for defects in the PCCA and PCCB genes encoding propionyl CoA carboxylase subunits. AB - Propionic acidemia can result from mutations in the PCCA or PCCB genes encoding the alpha and beta subunits, respectively, of propionyl-CoA carboxylase. We have developed a method based on complementation of the enzyme defect using a lipid mediated transient transfection of the normal human PCCA or PCCB cDNA into primary fibroblasts. We demonstrate the reliability of this method for identification of the defective PCC gene in order to unequivocally approach the mutational analysis in the corresponding PCCA and PCCB genes. PMID- 11914041 TI - Demonstration of altered splicing with the IVS3-1G --> a mutation of cathepsin C. AB - Papillon-Lefevre syndrome is an autosomal recessive palmoplantar keratoderma caused by cathepsin C gene mutations. We present the second family segregating the IVS3-1G --> A mutation and demonstrate for the first time that altered splicing and decreased enzymatic activity occur. RNA analysis revealed two species in carriers, corresponding to wild-type and mutant transcripts, and only the mutant transcript in affected individuals. Sequencing of the mutant transcript revealed that it lacked exon 3, resulting in a frameshift and introduction of a premature termination codon. PMID- 11914042 TI - Mental illness in mild PKU responds to biopterin. AB - A 25-year-old woman with mild hyperphenylalaninemia developed disabling depression and panic attacks. The mutations on the phenylalanine hydroxylase gene indicated that she might be responsive to tetrahydrobiopterin therapy. Mutation analyses were performed by the John F. Kennedy Institute in Glostrup, Denmark. The response to tetrahydrobiopterin therapy was impressive at an oral dose of 50 mg twice a day. A 25-year-old woman with mild hyperphenylalaninemia due to a PAH mutation of IVS12nt1g-->a/E390G has been treated for 1 year with BH4 therapy. A maintenance dosage of only 100 mg/day has resulted in significant improvement of depression and panic attacks, with discontinuation of psychotropic medication. PMID- 11914043 TI - IPF-1 gene variation and the development of type 2 diabetes. AB - Insulin promotor factor-1 (IPF-1) is a gene critical for pancreatic development and insulin transcription. We genotyped U.S. Caucasians with (n = 217) and without (n = 176) Type 2 diabetes to determine if three previously identified variants (Cys18Arg, Asp76Asn, Arg197His) in the IPF-1 gene play a role in the development of Type 2 diabetes. The Cys18Arg and Arg197His variants were not present in any subjects. The Asp76Asn variant was found in one control subject (0.9%) and none of the diabetic subjects. Similarly, none of the variants were detected in African American subjects with (n = 78) and without (n = 82) Type 2 diabetes. In conclusion, these variants are rare or absent in U.S. Caucasians and African Americans and therefore, unlikely to play a significant role in the development of Type 2 diabetes in these populations. PMID- 11914044 TI - The hemochromatosis N144H mutation of SLC11A3 gene in patients with type 2 diabetes. PMID- 11914045 TI - The self-reported likelihood of patient delay in breast cancer: new thoughts for early detection. AB - BACKGROUND: Delayed presentation of self-discovered breast symptoms influences stage of cancer at diagnosis and decreases breast cancer survival. METHODS: A total of 699 asymptomatic women (black, white, and Latino), recruited in community settings and stratified by age, income, and educational level, were surveyed for their likelihood to delay (J-Delay scale) in the event of a breast symptom discovery. Models of likelihood were tested with logistic regression analyses. RESULTS: A total of 166 women (23.7%) reported likelihood to delay. Lower income, lower educational level, self identification as Latino or black, experienced prejudice in care delivery, perceived lack of access to health care, fatalism about breast cancer, poor health care utilization habits, self-care behavior, spouse/partner and employer perceived constraints, problem-solving style, and a lack of knowledge of breast cancer's presenting symptoms were associated with likelihood to delay. A combined sample multiple logistic regression model correctly predicted 40.6% of women reporting a likelihood to delay, 94.9% of those not likely to delay, and 82.4% (551 of 669) of cases overall. CONCLUSIONS: Self-reported likelihood of patient delay is measurable in advance of symptom occurrence, and this measure is consistent with behavioral and knowledge variables previously linked with advanced breast cancer at diagnosis. PMID- 11914046 TI - Page for patients. Allergy season. PMID- 11914047 TI - Osteoporosis prevention: pediatricians' knowledge, attitudes, and counseling practices. AB - BACKGROUND: Strategies to prevent adult osteoporosis are best undertaken during childhood and adolescence, when the greatest amount of bone mineral density is acquired. This study examines pediatricians' knowledge and practices regarding osteoporosis prevention. METHODS: One hundred eighty-seven primary care pediatricians from San Diego and Imperial Counties responded to a 44-item mailed survey that measured physician knowledge, attitudes, and practices regarding osteoporosis prevention. RESULTS: Knowledge of U.S. RDA for calcium in children and adolescents was limited, with only 23.7 and 32.3% of pediatricians correctly identifying the recommended values for children and adolescents, respectively. Thirty-eight percent of respondents regarded osteoporosis prevention to be an important issue, and less than half reported that they counseled patients for osteoporosis prevention. Of those pediatricians who did perform osteoporosis counseling, the two most frequently cited motivators for counseling were that counseling is recommended by professional boards and having a personal or professional interest in osteoporosis prevention. The most commonly reported barriers to counseling were other issues taking greater priority and having insufficient time to undertake counseling for osteoporosis prevention. CONCLUSIONS: Osteoporosis prevention should begin in childhood and adolescence. This study suggests that many pediatricians do not counsel patients to reduce osteoporosis risk, nor do they consider this an important issue. Furthermore, they may not have full knowledge about how to prevent osteoporosis. PMID- 11914048 TI - What do middle school children bring in their bag lunches? AB - BACKGROUND: Limited documentation exists on nutritional characteristics of bag lunches, although about 30% of adolescents take them. This study describes: (a) the prevalence of different types of foods in bag lunches, (b) the number of kilocalories, percentage of kilocalories from fat, amount of total fat, saturated fat, cholesterol, and sugar, and (c) the differences related to gender and grade. METHODS: In 24 middle schools, observations were made on 1,381 bag lunches (about 58 per school) during one semester. RESULTS: The most common bag lunch components were beverages and sandwiches. Fruits were more common than vegetables; non-chip snacks and chips were more common than cookies, candy, and cakes/pies. Bag lunches averaged 596.2 kcal (29.7% from fat), 20.8 g of total fat, 6.2 g of saturated fat, 32.6 mg of cholesterol, and 21.3 g of sugar. Boys' bag lunches had significantly more kilocalories, total fat, saturated fat, cholesterol, and percentage of energy from fat than girls' lunches. Significant grade differences were found only for total fat and cholesterol, with seventh graders bringing more than sixth graders. CONCLUSION: More research on the nutritional quality of foods brought to school in bag lunches is important for gaining a better understanding about how to improve students' nutrition. PMID- 11914049 TI - Greek Christian Orthodox Ecclesiastical lifestyle: could it become a pattern of health-related behavior? AB - BACKGROUND: Although past research has globally supported the salutary impact of religion on health and health-related behaviors, it has not extensively examined the impact of the Christian Orthodox Church's way of living on people's health and health-related behavior. METHODS: Semistructured personal interviews were used to investigate a stratified sample of 20- to 65-year-old individuals in the greater Athens area. Constructs were compared to single items and indices, which varied across data sets. RESULTS: Multiple-regression analysis specify that persons adopting the Christian Orthodox Church's lifestyle were more likely to behave in ways that enhance their health (e.g., relaxation, life satisfaction, healthful nutrition, personal hygiene, and physical activity), after controlling for a set of socio-demographic factors and their current health status. CONCLUSION: These results suggest that the Christian Orthodox Church's lifestyle constitutes a pattern of health-related behavior. PMID- 11914050 TI - Lung cancer risk among Czech women: a case-control study. AB - BACKGROUND: Few data are available to explain the ongoing increase in lung cancer mortality among Czech women. The study is aimed at examining the role of smoking and known or suspected cofactors. METHODS: Data collected by in-person interviews from 269 female lung cancer cases and 1079 controls were analyzed using unconditional logistic regression and other methods. RESULTS: Cigarette smoking was the most important factor associated with excess risk of lung cancer among women. Risk was increased both among current smokers (OR = 10.30), long-term ex smokers (> or =10 years ago; OR = 3.79), and short-term ex-smokers (<10 years ago; OR = 14.63), all compared against never-smokers. In addition, significant associations with risk were found for chronic cough, chronic phlegm of less than 2-year duration, and shortness of breath. Inverse associations emerged for physical exercise and body mass index. Excess risk associated with consumption of red meat and poultry, and protective effects associated with intake of vegetables were restricted to squamous-, small-, and large-cell cancers combined, but were not apparent for adenocarcinoma. CONCLUSIONS: While smoking has been verified to be the main determinant of lung cancer risk among Czech women, cofactors such as diet, history of lung disease, and lifestyle factors may have a contributory role. PMID- 11914051 TI - Missed opportunities: intimate partner violence in family practice settings. AB - BACKGROUND: For women experiencing partner violence, women health care visits represent opportunities for physicians and patients to address intimate partner violence (IPV), a significant health threat for women. OBJECTIVES: The objectives were to estimate rates of physician documentation of IPV in medical records; characterize IPV+ women most likely to have IPV documented; and determine whether IPV screening increased IPV documentation. METHODS: Subjects were women ages 18 65 receiving primary care in two large family practice clinics. All were screened for IPV by study staff using a modified Index of Spouse Abuse and the Women's Experience with Battering scales. We selected and abstracted medical records for all women experiencing current IPV (N = 144) and a random sample of women never experiencing IPV (N = 147). RESULTS: Of 144 women screened as currently experiencing IPV, 14.7% were documented. Women most likely to have IPV documented were Caucasian, with higher WEB scores, and more likely to have an event that could trigger posttraumatic stress syndrome. Although the majority (41/56) of women currently in physically violent relationships did not plan to disclose IPV, those disclosing were significantly more likely to have IPV documented and documentation occurred after screening for 60% of women experiencing IPV. CONCLUSION: IPV screening increased documentation. IPV screening can provide the opportunity for patients to disclose IPV. Physicians then have the opportunity to compassionately connect patients with appropriate resources. PMID- 11914052 TI - Risk factors for hospitalized upper or lower gastrointestinal tract bleeding in treated hypertensives. AB - BACKGROUND: We examined risk factors for hospitalized gastrointestinal bleeding among health maintenance organization (HMO) members with hypertension. METHODS: Case subjects (n = 199) were patients with hypertension hospitalized for confirmed gastrointestinal bleeding in 1992-1994. Control hypertensive subjects (n = 821) were selected from ongoing studies. Medical records and computerized pharmacy data were used to assess risk factors. Adjusted relative risks (RRs) were estimated using logistic regression models. RESULTS: In multivariate adjusted models, significant risk factors for upper gastrointestinal bleeding (n = 111 cases) were hepatic disease (RR = 2.85), elevated creatinine (RR = 2.45), nonsteroidal anti-inflammatory drug use (RR = 2.28), smoking (RR = 1.93), cardiovascular disease (RR = 1.89), and physical inactivity (RR = 1.70). Risk factors for lower gastrointestinal bleeding (n = 43 cases) in multivariate adjusted analyses were anticoagulant or thrombolytic therapy (RR = 3.80), elevated creatinine (RR = 2.31), and physical inactivity (RR = 2.10). CONCLUSIONS: This study confirmed several known risk factors for hospitalized gastrointestinal bleeding, including hepatic disease, renal dysfunction, and medication use, and also identified smoking and physical inactivity as independent risk factors. The magnitude of the relative risks associated with these behavioral factors suggests that lifestyle modification may substantially reduce the risk of gastrointestinal bleeding. PMID- 11914053 TI - Quality of clinical reports on behavioral interventions for hypertension. AB - PURPOSE: The purpose of this study was to provide clinicians with the confidence to employ behavioral interventions for hypertension and to guide researchers in the development and reporting of studies. METHODS: We systematically reviewed English language articles (1970-1999) describing behavioral interventions for hypertension, evaluating aspects of design, analysis, reporting of results, and factors that were associated with higher quality of these studies. RESULTS: Of 100 articles, 49 were randomized controlled trials (RCT), 33 were observational studies with control groups, and 18 were observational studies without control groups; mean (SE) quality scores were 69.2 (1.6), 57.6 (5.3), and 60.3 (2.2), respectively. RCTs were more likely than observational studies to attain high scores in descriptions of appropriateness of control group, inclusion and exclusion criteria, study population, and the intervention protocol. In multivariate analysis, date of publication, reported funding source, and intervention type were independently associated with greater quality scores: 7.4 [95% CI: 0.03, 14.7] points greater for articles published 1990-1999 vs 1970 1979, 6.5 [95% CI: 1.4, 11.6] points greater for articles reporting government funding vs those not reporting funding sources, and 8.6 [95% CI: 0.3, 17.1], 12.9 [95% CI: 3.4, 22.4], and 14.2 [4.1, 24.4] points greater for articles examining patient education/support, change in delivery system, and mass health campaigns vs articles examining patient reminders, respectively. CONCLUSIONS: While quality has improved over time, there is considerable room for improvement. Investigators should pay particular attention to description of study population and allocation of subjects, the use of standardized outcomes reporting, and appropriate statistical analysis. PMID- 11914054 TI - Peer influences and access to cigarettes as correlates of adolescent smoking: a cross-cultural comparison of Wuhan, China, and California. AB - BACKGROUND: Few studies have assessed the effects of access to cigarettes and peer influences on adolescent smoking in non-Western countries. Using samples characterized by two distinct cultural, social, and economic systems, this study evaluated the associations of friends' smoking and perceived access to cigarettes with adolescent smoking behavior in California and Wuhan, China. METHODS: Survey data were obtained from 5870 eighth-grade students in the Independent Evaluation of the California Tobacco Control Program and 6992 seventh- to ninth-grade students in the Wuhan Smoking Prevention Trial. Odds ratios for lifetime and 30 day smoking, according to friends' smoking and perceived access to cigarettes, were calculated for boys and girls in both samples and compared. RESULTS: California students were more likely than Wuhan students to have friends who smoked and to perceive easy access to cigarettes. The smoking prevalence was lower in Wuhan than in California, mainly due to the low smoking prevalence among Wuhan girls. Friends' smoking was strongly associated with smoking in both samples, and the strength of this association did not differ between the two cultures. Access to cigarettes was associated with a higher risk of lifetime smoking in both cultures and a higher risk of past 30-day smoking in California only. CONCLUSIONS: Despite divergent tobacco control policy enforcement, social structures, and cultural contexts, similarities exist between Wuhan and California. The findings suggest support for adapting a social-influences-based smoking prevention program developed in the United States to the culturally specific needs of youth in Wuhan, China. PMID- 11914055 TI - Knowledge of osteoporosis in a Swedish municipality--a prospective study. AB - BACKGROUND: As a part of the Vadstena Osteoporosis Prevention Project, the knowledge of osteoporosis was examined before the intervention program started, after 5 and 10 years. METHODS: At baseline (in 1989) 15% of the population in two Swedish municipalities was randomly invited to the study. The participants in the study group were invited for examination by forearm bone densitometry and a questionnaire concerning lifestyle and risk factors for osteoporosis and also knowledge of osteoporosis, while the subjects in the control group were examined only by questionnaire. Follow-ups were made in 1994 and in 1999. Meanwhile education about osteoporosis was given to the study group, to the public, and to various professionals in the study community. RESULTS: There was a difference in the level of knowledge between the groups prior to the intervention. The rate of increment did not differ significantly between the groups for the study period. Previous participants had 0.58 higher score than new participants in the study group in 1994 (P = 0.031) and 0.76 higher score in 1999 (P < 0.001) regarding the total number of correct answers. The women in the study group had 0.63 higher score than the men in 1994 (P = 0.016) and 1.03 higher score in 1999 (P < 0.001) regarding the total number of correct answers. CONCLUSION: There was no significant effect of a general intervention program concerning the knowledge of osteoporosis in participants in the intervention area compared to the control area. PMID- 11914056 TI - A variable number of tandem repeats result in polymorphic alpha -isopropylmalate synthase in Mycobacterium tuberculosis. AB - A locus of variable number of the tandem repeat, VNTR4155, resides in the putative leuA gene, encoding for alpha -isopropylmalate synthase (alpha -IPMS) of Mycobacterium tuberculosis, a repeat that is unique to the bacterium. The objective was to determine whether the polymorphic VNTR4155 was translated and resulted in a polymorphic protein. The putative leuA gene of the M. tuberculosis H37Rv strain was cloned by PCR and expressed in a His-tagged form in Escherichia coli. The enzymatic properties of the purified protein were studied. The protein was used as an antigen to immunize rabbits. Soluble proteins of several strains of M. tuberculosis were examined by Western blot analysis. The polymorphism of VNTR4155 was due to the presence of different copy number of the 57-bp tandem repeat. The putative alpha -IPMS of various strains of M. tuberculosis had different sizes, varying directly with the length of their VNTR4155. PMID- 11914058 TI - Increased protection against bovine tuberculosis in the brushtail possum (Trichosurus vulpecula) when BCG is administered with killed Mycobacterium vaccae. AB - SETTING: The Australian brushtail possum is the major wildlife reservoir for Mycobacterium bovis infection in New Zealand. Development of an effective tuberculosis vaccine for possums will reduce the spread of infection to cattle and farmed deer. OBJECTIVES: To determine whether killed M. vaccae can improve the efficacy of vaccination with M. bovis bacillus Calmette Guerin (BCG) against bovine tuberculosis in the possum. DESIGN: Groups of possums (n=6-8) were vaccinated via intranasal and intraconjunctival routes with BCG alone or BCG in combination with heat-killed M. vaccae. Controls were non-vaccinated or vaccinated with heat-killed M. vaccae alone. After challenge with virulent M. bovis, protection was assessed by a reduction in loss of body weight and bacterial counts in lungs and spleens. Blood lymphocyte proliferative responses to M. bovis purified protein derivative were monitored throughout. RESULTS: The earliest lymphocyte responses following vaccination were from animals inoculated with BCG plus 100 microg heat-killed M. vaccae. Loss of body weight was significantly reduced in all BCG-vaccinated groups compared control groups. Spleen bacterial counts were significantly lower in animals vaccinated with M. vaccae plus BCG compared to the non-vaccinated group. Furthermore, vaccination with 100 microg M. vaccae plus BCG significantly reduced spleen bacterial counts compared to vaccination with BCG alone. CONCLUSION: The possum infection model is one of the first to show that novel vaccine strategies may offer better protection against tuberculosis than BCG alone. PMID- 11914057 TI - Decreased IFN- gamma and increased IL-4 production by human CD8(+) T cells in response to Mycobacterium tuberculosis in tuberculosis patients. AB - To investigate the role of MHC class I restricted CD8(+) T cells in host defense to M. tuberculosis, peripheral blood mononuclear cells (PBMC) from healthy BCG vaccinated donors and untreated pulmonary tuberculosis (TB) patients in The Gambia were stimulated for 6 days with M. bovis BCG or M. tuberculosis and the CD8(+) T cell response analyzed. Intracellular FACS analysis of cytokine production by CD8(+) T cells showed that IFN- gamma and TNF- alpha production were greatly reduced in TB patients compared to healthy controls. IL-4-producing CD8(+) T cells were detected in TB patients, a phenotype absent in controls. Collectively, these data suggest that an alteration in the type 1/type 2 cytokine balance occurs in CD8(+) T cells during clinical tuberculosis, and that this may provide a surrogate marker for disease. PMID- 11914059 TI - Dialogue between Mycobacterium tuberculosis and Homo sapiens Part II A long-term truce between multidrug-resistant mycobacteria and a TB patient. PMID- 11914060 TI - Interaction of Mycobacterium tuberculosis with human respiratory epithelial cells (HEp-2). AB - SETTING: Mycobacterium tuberculosis (MTB) is known to adhere to, invade, multiply inside, and modulate respiratory epithelial cell functions. However, the mode and the molecules involved in the interaction of MTB with respiratory cells are not known. OBJECTIVE: To study post-adherence morphological changes in human respiratory epithelial cells and identify MTB components involved in the binding to these cells. DESIGN: The mechanism of interaction of MTB human respiratory epithelial cells (HEp-2) was investigated by incubating MTB with HEp-2 cells, whereupon the morphological changes were examined under scanning electron microscopy. MTB components involved in the interaction with HEp-2 cells were identified by probing Western blots of mycobacterial sonicates with biotinylated HEp-2 extracts. RESULTS: MTB readily bound to the surface of HEp-2 cells. Adherence of tubercle bacilli on the surface caused membrane perturbation resulting in increased surface membrane projections, particularly towards the margins of the cells. Long thin epithelial membrane projections formed loops around the bacilli holding them on the surface. Membrane projections fused to form a web in which the bacilli were entrapped. Concomitant membrane cavitations seen beneath the site of attachment of the bacilli conceivably caused internalization of the organisms. At least five MTB proteins ( approximately 112, 35, 28, 21 and 15 kDa) specific for HEp-2 cells were identified. Of these, the 28 kDa protein was the predominant HEp-2-binding protein and was found to be heparin binding hemagglutinin (HBHA). While M. bovis BCG strain showed the same five HEp 2-binding proteins, M. smegmatis, displayed only one (31 kDa) HEp-2-specific protein. CONCLUSION: MTB binds to HEp-2 cells through multiple proteins, and the binding causes membrane perturbations that result in internalization of the organisms. PMID- 11914061 TI - Flow-cytometric assessment of lymphocyte cytokine production in tuberculosis. AB - We assessed by flow-cytometry the Th1/Th2 profiles in peripheral blood lymphocytes (PBL) from patients with active tuberculosis (TB), before and after antituberculous therapy, and from healthy tuberculin-positive and -negative reactors. PBL from patients showed a reduced potential for Th1-cytokine (notably IFN- gamma) production after culture with a policlonal stimulus. When these PBL from patients were cultured with a M. tuberculosis (MTB)-specific antigen such as PPD (10 microg/ml), there was no detectable production of Th1 cytokines. Only the Th2 cytokine IL10 was detected in PBL from patients but not from controls. However, at the site of the tuberculosis disease, T lymphocytes from bronchoalveolar lavage, after culture with PPD, produced IFN- gamma. After completion of tuberculosis therapy, PBL did not produce IL10. These data indicate that the immunosuppression observed in PBL during active tuberculosis infection may be related to IL10 production, and to the compartmentalization of the antigen Th1 response to sites of active MTB infection. PMID- 11914062 TI - The alternating site, binding change mechanism for proton translocation by transhydrogenase. PMID- 11914063 TI - Proteolytic DNA for mapping protein-DNA interactions. AB - We describe a technique to determine sites on proteins involved in protein-DNA interactions. DNA was synthesized via polymerase chain reaction (PCR) to produce four polynucleotide products with phosphorothioate nucleotides at the A, T, G, or C residues. Limited conjugation with the chemical protease FeBABE results in the surface of DNA being randomly labeled at the phosphorothioate sites with this protein-cleaving reagent. After formation of a protein-DNA complex, the proteolytic DNA can be activated to cleave the protein backbone at sites near the DNA. This technique was used to study the bacterial RNA polymerase/lacUV5 DNA open promoter complex, about which significant structural information is available. Cleavage sites on the two largest subunits of RNA polymerase, beta and beta', agree well with a recent model based on the crystal structure of the core enzyme alpha(2)betabeta' [Naryshkin, N., Revyakin, A., Kim, Y., Mekler, V., and Ebright, R. H. (2000) Cell 101, 601-611]. The cleavage site present on alpha supports previous studies regarding DNA binding regions of the alpha subunit. Cleavage sites identified throughout the sigma(70) subunit help to orient it with respect to the open promoter complex. PMID- 11914064 TI - Role of dimerization in KH/RNA complexes: the example of Nova KH3. AB - The K homology module, one of the most common RNA-binding motifs, is present in multiple copies in both prokaryotic and eukaryotic regulatory proteins. Increasing evidence suggests that self-aggregation of KH modules has a functional role. We have used a combination of techniques to characterize the behavior in solution of the third KH domain of Nova-1, a paradigmatic KH protein. The possibility of working on the isolated module allowed us to observe specifically the homodimerization and RNA-binding properties of KH domains. We provide conclusive evidence that self-association of Nova-1 KH3 occurs in solution even in the absence of RNA. Homodimerization involves a specific protein/protein interface. We also studied the dynamical behavior of Nova-1 KH3 in isolation and in complex with RNA. These data provide a model for the mechanism of KH/RNA recognition and suggest functional implications of dimerization in KH complexes. We discuss our findings in the context of the whole KH family and suggest a generalized mode of interaction. PMID- 11914065 TI - Determinants in the sequence specific binding of two plant transcription factors, CBF1 and NtERF2, to the DRE and GCC motifs. AB - Arabidopsis ERF proteins such as DREB1, DREB2, and CBF1 bind to the dehydration responsive element (DRE), which has the sequence TACCGACAT. Mutation analyses reveal that a central 5 bp CCGAC core of the DRE is the minimal sequence motif (designated as the DRE motif in this paper), to which the ERF domain fragment of CBF1 (CBF1-F) binds specifically with a binding K(d) at the nanomolar level. In contrast, the ERF domain fragment of the tobacco ERF2 (NtERF2-F) does not interact with the DRE motif, but restrictedly recognizes the sequence containing a minimal 6 bp GCCGCC motif (designated as the GCC motif in this paper). However, CBF1-F binds to the GCC motif with a binding activity similar to its binding activity for the DRE motif. These in vitro binding variations were further demonstrated through reporter cotransformation assays, suggesting that the DRE and GCC motifs are two similar sequence motifs sharing a common core region of CCGNC with a discriminating guanine base at the 5'-end of the GCC motif. Binding analyses with the mutated ERF domain show that such a unique binding of NtERF2-F to the GCC motif can be altered by the substitution of A14 with valine in beta strand 2 of its ERF domain, the mutant NtERF2-F, ERFav, acquiring a binding to the DRE motif with a K(d) comparable to that for CBF1-F binding to the DRE motif. This demonstrates that A14 is an important determinant of the NtERF2-F binding specificity. A possible mechanism of the binding specificity determination is discussed. PMID- 11914066 TI - Mutations of tyrosine 537 in the human estrogen receptor-alpha selectively alter the receptor's affinity for estradiol and the kinetics of the interaction. AB - Mutation of tyrosine 537 (Y537) of the human estrogen receptor-alpha (hERalpha) produces receptors having a range of constitutive activity, which suggests that this residue modulates the conformational changes of the receptor. We investigated the effect of several mutations at this position, to phenylalanine (Y537F), to serine (Y537S), and to glutamic acid (Y537E), on the hormone-binding properties of the receptor. The affinities of the wt, the Y537F mutant, and the Y537S mutant for estradiol were similar: K(a) = 2.2 +/- 0.2, 3.9 +/- 0.5, and 2.8 +/- 0.4 nM(-1), respectively. By contrast, the affinity of the Y537E mutant for estradiol was reduced 10-fold, K(a) = 0.2 +/- 0.1 nM(-1). All proteins bound [(3)H]estradiol with a positive cooperative mechanism (n(H) = 1.7-1.9), indicating they can form dimers. The wt receptor and the Y537S and Y537E mutants exhibited biphasic dissociation kinetics, which is also indicative of dimerization. Surprisingly, the half-lives of the slow component of the wt and the Y537E mutant were indistinguishable, 118 +/- 3.4 and 122 +/- 4.5 min, respectively, even though the affinity of the Y537E mutant for hormone was reduced 10-fold. The half-life of the slow component of the Y537S mutant was reduced to 96.5 +/- 3.8 min. Molecular models were constructed and compared to identify changes in the structure that correlate with the observed effects on hormone binding. Local alterations in hydrogen bonding, the position of side chains, and the position of the peptide backbone were observed. Taken together, these results show that mutations at Y537 selectively alter the affinity and kinetics of hormone binding to the receptor, and are consistent with the idea that the estradiol-estrogen receptor interaction can follow more than one pathway. PMID- 11914067 TI - Antibiotic resistance peptides: interaction of peptides conferring macrolide and ketolide resistance with Staphylococcus aureus ribosomes: conformation of bound peptides as determined by transferred NOE experiments. AB - Two antibiotic resistance peptides, the E-peptide (MRLFV) and the K-peptide (MRFFV) conferring macrolide and ketolide resistance, respectively, were studied in the complex state with bacterial Staphylococcus aureus ribosomes. Interactions of antibiotic resistance peptides with ribosomes were investigated using two dimensional transferred nuclear Overhauser effect spectroscopy (TRNOESY), suggesting that the peptide-ribosome interaction was associated with the low affinity binding level. K-Peptide displayed a significantly better response in TRNOEs NMR experiments, in agreement with a better overall antibiotic activity of ketolides. This difference highlights a mimetic effect displayed by the E- and K peptides. This study shows that conformation plays an essential role for the affinity binding site and, thus, for the resistance mechanism. Specific conformations were preferred in the bound state; their superimposition exhibited a similar cyclic peptidyl chain, while the side chain region varies. The F4 phenyl moiety in E-peptide has moved out of the turn region compared to its folding in the ketolide resistance peptide. In the K-peptide binding surface, the F4 aromatic chain is maintained by stacking with the guanidyl group of the R2 residue providing a particular hydrophobic and globular fragment, which may be important for the ketolide resistance peptide mode of action. Additionally, T(2) (CPMG) measurements were used to characterize equilibrium binding of antibiotic resistance peptides to bacterial ribosomes. The results bring to the fore E- and K-peptide competition with antibiotics for binding to the ribosomes. Their specific interaction and their competitive effects reveal a novel aspect of interaction of resistance peptides with ribosomes and suggest new insights about their mode of action. The resistance mechanism may imply two steps, a competitive effect of the resistance peptide for the macrolide (or ketolide) binding site followed by a "bottle brush" effect in which the drug and the peptide are driven out their binding site on the ribosome. PMID- 11914068 TI - Catalysis and stability of triosephosphate isomerase from Trypanosoma brucei with different residues at position 14 of the dimer interface. Characterization of a catalytically competent monomeric enzyme. AB - In homodimeric triosephosphate isomerase from Trypanosoma brucei (TbTIM), cysteine 14 of each the two subunits forms part of the dimer interface. This residue is central for the catalysis and stability of TbTIM. Cys14 was changed to the other 19 amino acids to determine the characteristics that the residue must have to yield catalytically competent stable enzymes. C14A, C14S, C14P, C14T, and C14V TbTIMs were essentially wild type in activity and stability. Mutants with Asn, Arg, and Gly had low activities and stabilities. The other mutants had less than 1% of the activity of TbTIM. One of the latter enzymes (C14F) was purified to homogeneity. Size exclusion chromatography and equilibrium sedimentation studies showed that C14F TbTIM is a monomer, with a k(cat) approximately 1000 times lower and a K(m) approximately 6 times higher than those of TbTIM. In C14F TbTIM, the ratio of the elimination (methylglyoxal and phosphate formation) to isomerization reactions was higher than in TbTIM. Its secondary structure was very similar to that of TbTIM; however, the quantum yield of its aromatic residues was lower. The analysis of the data with the 19 mutants showed that to yield enzymes similar to the wild type, the residue must have low polarity and a van der Waals volume between 65 and 110 A(3). The results with C14F TbTIM illustrate that the secondary structure of TbTIM can be formed in the absence of intersubunit contacts, and that it has sufficient tertiary structure to support catalysis. PMID- 11914069 TI - Calcium coordination studies of the metastatic Mts1 protein. AB - Mts1, also known as S100A4, is an 11 kDa calcium-binding protein strongly linked to metastasis. As a member of the S100 protein family, Mts1 is predicted to contain four alpha-helices and two calcium-binding loops, the second of which forms a canonical EF hand, while the first is a pseudo-EF hand, using two extra residues and principally backbone carbonyls rather than side chain oxygens to coordinate calcium. Here we follow chemical shift changes which occur in Mts1 upon titration of calcium. The results are consistent with calcium coordination by the EF hands described above. Filling of the first (pseudo) EF hand occurs at a lower calcium concentration than does filling of the second (canonical) EF hand. Concurrent with filling of site I, resonances from much of helix 4 vanish while the chemical shifts of a possibly nascent helical segment immediately C terminal to helix 4 increase in helical character. Other smaller changes are seen, including a change in the linker joining helix 2 and helix 3. Since binding of effector molecules to S100 proteins has been shown to involve the C-terminus and linker regions, these calcium-induced changes have implications for the role of Mts1 in metastasis. PMID- 11914070 TI - High-resolution crystal structures of the lectin-like xylan binding domain from Streptomyces lividans xylanase 10A with bound substrates reveal a novel mode of xylan binding. AB - Carbohydrate-binding module (CBM) family 13 includes the "R-type" or "ricin superfamily" beta-trefoil lectins. The C-terminal CBM, CBM13, of xylanase 10A from Streptomyces lividans is a family 13 CBM that is not only structurally similar to the "R-type" lectins but also somewhat functionally similar. The primary function of CBM13 is to bind the polysaccharide xylan, but it retains the ability of the R-type lectins to bind small sugars such as lactose and galactose. The association of CBM13 with xylan appears to involve cooperative and additive participation of three binding pockets in each of the three trefoil domains of CBM13, suggesting a novel mechanism of CBM-xylan interaction. Thus, the interaction of CBM13 with sugars displays considerable plasticity for which we provide a structural rationale. The high-resolution crystal structure of CBM13 was determined by multiple anomalous dispersion from a complex of CBM13 with a brominated ligand. Crystal structures of CBM13 in complex with lactose and xylopentaose revealed two distinct mechanisms of ligand binding. CBM13 has retained its specificity for lactose via Ricin-like binding in all of the three classic trefoil binding pockets. However, CBM13 has the ability to bind either the nonreducing galactosyl moiety or the reducing glucosyl moiety of lactose. The mode of xylopentaose binding suggests adaptive mutations in the trefoil sugar binding scaffold to accommodate internal binding on helical polymers of xylose. PMID- 11914071 TI - Site-specific characterization of the association of xylooligosaccharides with the CBM13 lectin-like xylan binding domain from Streptomyces lividans xylanase 10A by NMR spectroscopy. AB - Endo-beta-1,4-xylanase 10A (Xyn10A) from Streptomyces lividans includes an N terminal catalytic module and a 130-residue C-terminal family 13 carbohydrate binding module (CBM13). This latter domain adopts a beta-trefoil structure with three potential binding sites (alpha, beta, and gamma) for a variety of small sugars, xylooligosaccharides, and xylan polymers. To investigate the role of this multivalency in carbohydrate binding, we have used NMR spectroscopy to characterize the interaction of isolated CBM13 with a series of sugars. We have assigned resonances from the main chain nuclei of CBM13 using heteronuclear NMR experiments. Analysis of (15)N NMR relaxation data using the extended model free formalism reveals that CBM13 tumbles as an oblate ellipsoid (D( parallel)/D( perpendicular) = 0.80 +/- 0.02) and that its backbone is relatively rigid on the sub-nanosecond time scale. In particular, the three binding sites show no distinct patterns of increased internal mobility. Ligand-induced chemical shift changes in the (1)H-(15)N HSQC spectra of CBM13 were monitored as a function of increasing concentrations of L-arabinose, lactose, D-xylose, xylobiose, xylotetraose, and xylohexaose. Patterns of shift perturbations for well-resolved resonances demonstrate that all of these sugars associate independently with the three binding sites of CBM13. On the basis of the site-specific association constants derived from a quantitative analysis of these titration data, we show that L-arabinose, lactose, and D-xylose preferentially bind to the alpha site of CBM13, xylobiose binds equally well to all three sites, and xylotetraose and xylohexaose prefer binding to the beta site. Inspection of the crystallographic structure of CBM13 [Notenboom, V., Boraston, A. B., Williams, S. J., Kilburn, D. G., and Rose, D. R. (2002) Biochemistry 41, 4246-4254] provides a rationalization for these results. PMID- 11914072 TI - Crystallographic study of the recombinant flavin-binding domain of Baker's yeast flavocytochrome b(2): comparison with the intact wild-type enzyme. AB - Flavocytochrome b(2) catalyzes the oxidation of L-lactate to pyruvate and the transfer of electrons to cytochrome c. The enzyme consists of a flavin-binding domain, which includes the active site for lacate oxidation, and a b(2) cytochrome domain, required for efficient cytochrome c reduction. To better understand the structure and function of intra- and interprotein electron transfer, we have determined the crystal structure of the independently expressed flavin-binding domain of flavocytochrome b(2) to 2.50 A resolution and compared this with the structure of the intact enzyme, redetermined at 2.30 A resolution, both structures being from crystals cooled to 100 K. Whereas there is little overall difference between these structures, we do observe significant local changes near the interface region, some of which impact on amino acid side chains, such as Arg289, that have been shown previously to have an important role in catalysis. The disordered loop region found in flavocytochrome b(2) and its close homologues remain unresolved in frozen crystals of the flavin-binding domain, implying that the presence of the b(2)-cytochrome domain is not responsible for this positional disorder. The flavin-binding domain interacts poorly with cytochrome c, but we have introduced acidic residues in the interdomain interface region with the aim of enhancing cytochrome c binding. While the mutations L199E and K201E within the flavin-binding domain resulted in unimpaired lactate dehydrogenase activity, they failed to enhance electron transfer rates with cytochrome c. This is most likely due to the disordered loop region obscuring all or part of the surface having the potential for productive interaction with cytochrome c. PMID- 11914073 TI - Crystallographic evidence of a transglycosylation reaction: ternary complexes of a psychrophilic alpha-amylase. AB - The psychrophilic Pseudoalteromonas haloplanctis alpha-amylase is shown to form ternary complexes with two alpha-amylase inhibitors present in the active site region, namely, a molecule of Tris and a trisaccharide inhibitor or heptasaccharide inhibitor, respectively. The crystal structures of these complexes have been determined by X-ray crystallography to 1.80 and 1.74 A resolution, respectively. In both cases, the prebound inhibitor Tris is expelled from the active site by the incoming oligosaccharide inhibitor substrate analogue, but stays linked to it, forming well-defined ternary complexes with the enzyme. These results illustrate competition in the crystalline state between two inhibitors, an oligosaccharide substrate analogue and a Tris molecule, bound at the same time in the active site region. Taken together, these structures show that the enzyme performs transglycosylation in the complex with the pseudotetrasaccharide acarbose (confirmed by a mutant structure), leading to a well-defined heptasaccharide, considered as a more potent inhibitor. Furthermore, the substrate-induced ordering of water molecules within a channel highlights a possible pathway used for hydrolysis of starch and related poly- and oligosaccharides. PMID- 11914074 TI - Co- and posttranslational modification of the alpha(1B)-adrenergic receptor: effects on receptor expression and function. AB - We have characterized the maturation, co- and posttranslational modifications, and functional properties of the alpha(1B)-adrenergic receptor (AR) expressed in different mammalian cells transfected using conventional approaches or the Semliki Forest virus system. We found that the alpha(1B)-AR undergoes N-linked glycosylation as demonstrated by its sensitivity to endoglycosidases and by the effect of tunicamycin on receptor maturation. Pulse-chase labeling experiments in BHK-21 cells demonstrate that the alpha(1B)-AR is synthesized as a 70 kDa core glycosylated precursor that is converted to the 90 kDa mature form of the receptor with a half-time of approximately 2 h. N-Linked glycosylation of the alpha(1B)-AR occurs at four asparagines on the N-terminus of the receptor. Mutations of the N-linked glycosylation sites did not have a significant effect on receptor function or expression. Surprisingly, receptor mutants lacking N linked glycosylation migrated as heterogeneous bands in SDS-PAGE. Our findings demonstrate that N-linked glycosylation and phosphorylation, but not palmitoylation or O-linked glycosylation, contribute to the structural heterogeneity of the alpha(1B)-AR as it is observed in SDS-PAGE. The modifications found are similar in the different mammalian expression systems explored. Our findings indicate that the Semliki Forest virus system can provide large amounts of functional and fully glycosylated alpha(1B)-AR protein suitable for biochemical and structural studies. The results of this study contribute to elucidate the basic steps involved in the processing of G protein-coupled receptors as well as to optimize strategies for their overexpression. PMID- 11914075 TI - Epidermal growth factor contains both positive and negative determinants for interaction with ErbB-2/ErbB-3 heterodimers. AB - Epidermal growth factor (EGF) and transforming growth factor (TGF)-alpha are potent activators of the ErbB-1 receptor, but, unlike TGF-alpha, EGF is also a weak activator of ErbB-2/ErbB-3 heterodimers. To understand the specificity of EGF-like growth factors for binding to distinct ErbB members, we used EGF/TGF alpha chimeras to examine the requirements for ErbB-2/ErbB-3 activation. Here we show that in contrast to these two wild-type ligands, distinct EGF/TGF-alpha chimeras are potent activators of ErbB-2/ErbB-3 heterodimers. On the basis of differences in the potency of these various chimeras, specific residues in the linear N-terminal region and the so-called B-loop of these ligands were identified to be involved in interaction with ErbB-2/ErbB-3. A chimera consisting of human EGF sequences with the linear N-terminal region of human TGF-alpha was found to be almost as potent as the natural ligand neuregulin (NRG)-1beta in activating 32D cells expressing ErbB-2/ErbB-3 and human breast cancer cells. Binding studies revealed that this chimera, designated T1E, has high affinity for ErbB-2/ErbB-3 heterodimers, but not for ErbB-3 alone. Subsequent exchange studies revealed that introduction of both His2 and Phe3 into the linear N-terminal region was already sufficient to make EGF a potent activator of ErbB-2/ErbB-3 heterodimers, indicating that these two amino acids contribute positively to this receptor binding. Analysis of the B-loop revealed that Leu26 in EGF facilitates interaction with ErbB-2/ErbB-3 heterodimers, while the equivalent Glu residue in TGF-alpha impairs binding. Since all EGF/TGF-alpha chimeras tested have maintained high binding affinity for ErbB-1, it is concluded that the diversity of the ErbB signaling network is determined by specific amino acids that facilitate binding to one receptor member, in addition to residues that impede binding to other ErbB family members. PMID- 11914076 TI - Interactions of cytoplasmic dynein light chains Tctex-1 and LC8 with the intermediate chain IC74. AB - The interactions of three subunits of cytoplasmic dynein from Drosophila melanogaster, LC8, Tctex-1, and the N-terminal domain of IC74 (N-IC74, residues 1 289), were characterized in vitro by affinity methods, limited proteolysis, and circular dichroism spectroscopy. These subunits were chosen for study because they are presumed to promote the assembly of the complex and to be engaged in the controlled binding and release of cargo. Limited proteolysis and mass spectrometry of N-IC74 in the presence of LC8 and Tctex-1 localized binding of Tctex-1 to the vicinity of K104 and K105, and localized binding of LC8 to the region downstream of K130. Circular dichroism, fluorescence, sedimentation velocity, and proteolysis studies indicate that N-IC74 has limited secondary and tertiary structure at near physiological solution conditions. Upon addition of LC8, N-IC74 undergoes a significant conformational change from largely unfolded to a more ordered structure. This conformational change is reflected in increased global protection of N-IC74 from proteolytic digestion following the interaction, and in a significant change in the CD signal. A smaller but reproducible change in the CD spectra was observed upon Tctex-1 binding as well. The increased structure introduced into N-IC74 upon light chain binding suggests a mechanism by which LC8 and Tctex-1 may regulate the assembly of the dynein complex. PMID- 11914077 TI - Zn(2+) binding properties of single-point mutants of the C-terminal zinc finger of the HIV-1 nucleocapsid protein: evidence of a critical role of cysteine 49 in Zn(2+) dissociation. AB - The two highly conserved Zn(2+) finger motifs of the HIV-1 nucleocapsid protein, NCp7, strongly bind Zn(2+) through coordination of one His and three Cys residues. To further analyze the role of these residues, we investigated the Zn(2+) binding and acid-base properties of four single-point mutants of a short peptide corresponding to the distal finger motif of NCp7. In each mutant, one Zn(2+)-coordinating residue is substituted with a noncoordinating one. Using the spectroscopic properties of Co(2+), we first establish that the four mutants retain their ability to bind a metal cation through a four- or five-coordinate geometry with the vacant ligand position(s) presumably occupied by water molecule(s). Moreover, the pK(a) values of the three Cys residues of the mutant apopeptide where His44 is substituted with Ala are found by (1)H NMR to be similar to those of the native peptide, suggesting that the mutations do not affect the acid-base properties of the Zn(2+)-coordinating residues. The binding of Zn(2+) was monitored by using the fluorescence of Trp37 as an intrinsic probe. At pH 7.5, the apparent Zn(2+) binding constants (between 1.6 x 10(8) and 1.3 x 10(10) M(-)(1)) of the four mutants are strongly reduced compared to those of the native peptide but are similar to those of various host Zn(2+) binding proteins. As a consequence, the loss of viral infectivity following the mutation of one Zn(2+)-coordinating residue in vivo may not be related to the total loss of Zn(2+) binding. The pH dependence of Zn(2+) binding indicates that the coordinating residues bind Zn(2+) stepwise and that the free energy provided by the binding of a given residue may be modulated by the entropic contribution of the residues already bound to Zn(2+). Finally, the pK(a) of Cys49 in the holopeptide is found to be 5.0, a value that is at least 0.7 unit higher than those for the other Zn(2+)-coordinating residues. This implies that Cys49 may act as a switch for Zn(2+) dissociation in the distal finger motif of NCp7, a feature that may contribute to the high susceptibility of Cys49 to electrophilic attack. PMID- 11914078 TI - A multigeneration analysis of cytochrome b(562) redox variants: evolutionary strategies for modulating redox potential revealed using a library approach. AB - The redox potential of cytochromes sets the energy yield possible in metabolism and is also a key determinant of the rate at which redox reactions proceed. Here, the heme protein, cytochrome b(562), is used to study the in vitro evolution of redox potential within a library of variants containing the same structural archetype, the four-helix bundle. Multisite variations in the active site of cytochrome b(562) were introduced. A library of variants containing random mutations in place of R98 and R106 was created, and the redox potentials of a statistical sampling of this library were measured. This procedure was carried out for both the low- and high-potential variants of a previously studied F61X/F65X, first-generation library [Springs, S. L., Bass, S. E., and McLendon, G. L. (2000) Biochemistry 39, 6075]. The second-generation library reported here has a range of redox potentials which is greater than 40% (160 mV) of the known accessible potential among cytochromes with identical axial ligands (but different folds) and exceeds the range exhibited phylogenetically by the cytochrome c' family which internally maintains the same axial ligation and fold. A statistical analysis of the libraries examined reveals that the redox potential of WT cyt b(562) is found at the high-potential extremum of the distribution, indicating that this protein apparently evolved to differentially stabilize the reduced protein. The 2.7 A crystal structure of F61I/F65Y/R106L (low-potential variant of the second-generation library) was solved and is compared to the wild type structure and the 2.2 A resolution structure of the F61I/F65Y variant (low potential variant of the first-generation library). The structures indicate that charge-dipole effects are responsible for shifting the redox equilibrium toward the oxidized state in both the F61I/F65Y and F61I/F65Y/R106L variants. Specifically, a new protein dipole is introduced into the heme microenvironment as a result of the F65Y mutation, two new internal water molecules (one in hydrogen-bonding distance of Y65) are found, and in the case of F61I/F65Y/R106L (DeltaE(m) = 158 mV vs NHE), increased solvent exposure of the heme as a result of the R106L substitution is identified. PMID- 11914079 TI - Mechanism of inhibition of the beta-lactamase of Enterobacter cloacae P99 by 1:1 complexes of vanadate with hydroxamic acids. AB - The class C beta-lactamase of Enterobacter cloacae P99 is competitively inhibited by low concentrations of 1:1 complexes of vanadate and hydroxamic acids. Structure-activity studies indicated that the hydroxamic acid functional group was essential to this inhibition. Both aryl and alkyl hydroxamic acids form inhibitory ternary complexes with vanadate and the enzyme, although, in certain cases of the latter, the inhibition may not be seen because of the low formation constants of the vanadate-hydroxamic acid complex. After all of the vanadate species present in solution had been taken into account, "real" K(i) values for the vanadate complexes could be determined. The K(i) value of the best of the inhibitors that were investigated, the 1:1 complex of vanadate with 4 nitrobenzohydroxamic acid, was 0.48 microM. Kinetics studies showed that the association and dissociation rate constants of this complex with the enzyme were 1.48 x 10(6) s(-1) M(-1) and 0.73 s(-1), respectively; the magnitude of the latter indicates covalent interaction of the complex with the enzyme. (51)V NMR and UV-vis spectra suggest that the structure of the vanadate complex bound to the enzyme may be very similar to that in solution. A (13)C NMR spectrum of the enzyme complex with 4-nitrobenzo[(13)C]hydroxamic acid and vanadate yields a coordination-induced shift (CIS) of 7.74 ppm. This is significantly larger than that of the vanadate complex in free solution (3.62 ppm), suggesting either, somewhat contrary to the (51)V and UV-vis spectra, greater interaction between vanadium and the hydroxamate carbonyl oxygen in the enzyme complex than in free solution or, more likely, polarization of the hydroxamate by interaction, e.g., hydrogen bonding, with the enzyme. Molecular modeling indicates that a pentacoordinated vanadate complex may well be able to snugly occupy the enzyme active site; Asn 152 is suitably placed to hydrogen bond to the hydroxamic acid oxygen atom. The experimental results are in accord with a model whereby the vanadate-hydroxamate-enzyme complex is a moderately good analogue of the transition state of the reaction of the beta-lactamase with phosphonate inhibitors. PMID- 11914080 TI - The role of the specificity-determining loop of the integrin beta subunit I-like domain in autonomous expression, association with the alpha subunit, and ligand binding. AB - Integrin beta subunits contain a highly conserved I-like domain that is known to be important for ligand binding. Unlike integrin I domains, the I-like domain requires integrin alpha and beta subunit association for optimal folding. Pactolus is a novel gene product that is highly homologous to integrin beta subunits but lacks associating alpha subunits [Chen, Y., Garrison, S., Weis, J. J., and Weis, J. H. (1998) J. Biol. Chem. 273, 8711-8718] and a approximately 30 amino acid segment corresponding to the specificity-determining loop (SDL) in the I-like domain. We find that the SDL is responsible for the defects in integrin beta subunit expression and folding in the absence of alpha subunits. When transfected in the absence of alpha subunits into cells, extracellular domains of mutant beta subunits lacking SDL, but not wild-type beta subunits, were well secreted and contained immunoreactive I-like domains. The purified recombinant soluble beta1 subunit with the SDL deletion showed an elongated shape in electron microscopy, consistent with its structure in alphabeta complexes. The SDL segment is not required for formation of alpha5beta1, alpha4beta1, alphaVbeta3, and alpha6beta4 heterodimers, but is essential for fomation of alpha6beta1, alphaVbeta1, and alphaLbeta2 heterodimers, suggesting that usage of subunit interface residues is variable among integrins. The beta1 SDL is required for ligand binding and for the formation of the epitope for the alpha5 monoclonal antibody 16 that maps to loop segments connecting blades 2 and 3 of beta propeller domain of alpha5, but is not essential for nearby beta-propeller epitopes. PMID- 11914081 TI - Kinetics and mechanism of superoxide reduction by two-iron superoxide reductase from Desulfovibrio vulgaris. AB - Superoxide reductases (SORs) contain a novel square pyramidal ferrous [Fe(NHis)(4)(SCys)] site that rapidly reduces superoxide to hydrogen peroxide. Here we report extensive pulse radiolysis studies on recombinant two-iron SOR (2Fe-SOR) from Desulfovibrio vulgaris. The results support and elaborate on our originally proposed scheme for reaction of the [Fe(NHis)(4)(SCys)] site with superoxide [Coulter, E. D., Emerson, J. E., Kurtz, D. M., Jr., and Cabelli, D. E. (2000) J. Am. Chem. Soc. 122, 11555-11556]. This scheme consists of second-order diffusion-controlled formation of an intermediate absorbing at approximately 600 nm, formulated as a ferric-(hydro)peroxo species, and its decay to the carboxylate-ligated ferric [Fe(NHis)(4)(SCys)] site with loss of hydrogen peroxide. The second-order rate constant for formation of the 600-nm intermediate is essentially pH-independent (pH 5-9.5), shows no D(2)O solvent isotope effect at pH 7.7, and decreases with increasing ionic strength. These data indicate that formation of the intermediate does not involve a rate-determining protonation, and are consistent with interaction of the incoming superoxide anion with a positive charge at or near the ferrous [Fe(NHis)(4)(SCys)] site. The rate constant for decay of the 600-nm intermediate follows the pH-dependent rate law: k(2)(obs) = k(2)'[H(+)] + k(2)' ' and shows a significant D(2)O solvent isotope effect at pH 7.7. The values of k(2)' and k(2)' ' indicate that the 600-nm intermediate decays via diffusion-controlled protonation at acidic pHs and a first-order process involving either water or a water-exchangeable proton on the protein at basic pHs. The formation and decay rate constants for an E47A variant of 2Fe-SOR are not significantly perturbed from their wild-type values, indicating that the conserved glutamate carboxylate does not directly displace the (hydro)peroxo ligand of the intermediate at basic pHs. The kinetics of a K48A variant are consistent with participation of the lysyl side chain in directing the superoxide toward the active site and in directing the protonation pathway of the ferric-(hydro)peroxo intermediate toward release of hydrogen peroxide. PMID- 11914082 TI - Subcellular localization of chlorosome proteins in Chlorobium tepidum and characterization of three new chlorosome proteins: CsmF, CsmH, and CsmX. AB - Chlorosomes are unique light-harvesting structures found in two families of photosynthetic bacteria. In this study, three chlorosome proteins (CsmF, CsmH, and CsmX) of the green sulfur bacterium Chlorobium tepidum were characterized by cloning and sequencing the genes which encode them, by overproducing the respective proteins in Escherichia coli, and by raising polyclonal antisera to the purified proteins. Three other proteins (AtpF, CT1970, and CT2144) which were identified in chlorosome fractions have similarly been characterized. The antisera were used to establish the distribution of each protein in various cellular fractions. Ten chlorosome proteins (CsmA, CsmB, CsmC, CsmD, CsmE, CsmF, CsmH, CsmI, CsmJ, and CsmX) copurified in a constant proportion together with bacteriochlorophyll c, and none of these 10 proteins was found in substantial amounts in other subcellular fractions. An antiserum to CsmH was highly effective in agglutinating chlorosomes, and antisera to CsmI, CsmJ, CsmX, and CsmA also immunoprecipitated chlorosomes to varying extents. However, an antiserum to CsmF did not agglutinate chlorosomes. The sequences of chlorosome proteins generally are not significantly similar to the sequences of other proteins in the databases. However, the N-terminal domains of three chlorosome proteins, CsmI, CsmJ, and CsmX, are related to adrenodoxin-type ferredoxins that ligate [2Fe-2S] clusters [Vassilieva, E. V., Antonkine, M. L., Zybailov, B. L., Yang, F., Jakobs, C. U., Golbeck, J. H., and Bryant, D. A. (2001) Biochemistry 40, 464-473]. The sequences of the C-terminal domains of these three proteins appear to be distantly related to CsmA and CsmE. The remaining chlorosome proteins can be divided into two additional structural families, CsmB/F and CsmC/D. CsmH is recovered in water-soluble form after overproduction in E. coli. Interestingly, this protein contains an N-terminal domain that is similar to CsmB/D, while its C terminal domain is related to CsmC/D. The sequence relationships indicate that, although the protein composition of Chlorobium-type chlorosomes is superficially more complex than that of the chlorosomes of Chloroflexus aurantiacus, this heterogeneity is mostly produced by gene duplication and divergence among a small number of protein types. PMID- 11914083 TI - Cholate-induced dimerization of detergent- or phospholipid-solubilized bovine cytochrome C oxidase. AB - Bovine heart cytochrome c oxidase (CcO), solubilized by either nonionic detergents or phospholipids, completely dimerizes upon the addition of bile salts, e.g., sodium cholate, sodium deoxycholate, or CHAPS. Bile salt induced dimerization occurs whether dodecyl maltoside, decyl maltoside, or Triton X-100 is the primary solubilizing detergent or the enzyme is dispersed in phosphatidylcholine, phosphatidylethanolamine, or mixtures thereof. In each case, complete CcO dimerization can be verified by sedimentation velocity and sedimentation equilibrium after correction for bound detergent and/or phospholipid. The relative concentration of the bile salt is critical for production of homogeneous, dimeric CcO. For example, enzyme solubilized by 2 mM detergent requires an equal molar concentration of sodium cholate. Similarly, enzyme dispersed in 20 mM phospholipid requires 50 mM sodium cholate, concentrations that are commonly used to reconstitute CcO into small unilamellar vesicles. Bile salts do more than just stabilize dimeric CcO and prevent detergent-induced dissociation into monomers. They are able to completely reverse detergent-induced monomerization and cause completely monomeric CcO to reassociate. Dimeric CcO so generated is no more stable than the original complex and easily dissociates into monomers if the bile salt is removed. The dimerization process is dependent upon a full complement of subunits; e.g., if subunits VIa and VIb are removed, the resulting monomeric CcO will not reassociate upon the addition of sodium cholate. These results support four important consequences: (1) dissociation of dimeric CcO into monomers is reversible; (2) stable dimers can be produced under solution conditions; (3) dimers can be stabilized even at relatively high pH and low enzyme concentration; and (4) subunits VIa and VIb are required for dimerization. PMID- 11914085 TI - The role of a parasite-specific allosteric site in the distinctive activation behavior of Eimeria tenella cGMP-dependent protein kinase. AB - A cGMP-dependent protein kinase (PKG) was recently identified as an anticoccidial target for the apicomplexan parasite Eimeria tenella [Gurnett, A., Liberator, P. A., Dulski, P., Salowe, S., Donald, R. G. K., Anderson, J., Wiltsie, J., Diaz, C., Harris, G., Chang, B., Darkin-Rattray, S. J., Nare, B., Crumley, T., Blum, P., Misura, A., Tamas, T., Sardana, M., Yuan, J., Biftu, T., and Schmatz, D. (2002) J. Biol. Chem. (in press)]. Unlike the PKGs of higher organisms that have two cGMP binding sites in their regulatory domain, the PKG from Eimeria tenella (Et-PKG) contains three putative cGMP binding sites and has distinctive activation properties, including a very large stimulation by cGMP ( approximately 1000-fold) with significant cooperativity (Hill coefficient of 1.7). During our investigation of Et-PKG activation, we found that 8-substituted cGMP analogues are weak partial activators. For example, 8-NBD-cGMP provides a maximal stimulation of activity of only 20-fold with little evident cooperativity, although cGMP can synergize with the analogue to provide full activation. The results suggest that partial activation is a consequence of restricted binding of 8-NBD-cGMP to a subset of cGMP sites in the enzyme. Site-directed mutagenesis of conserved arginine and glutamate residues in the parasite-specific third cGMP site confirms that this site is an important functional participant in the allosteric regulation of the kinase and that it exhibits very high selectivity against 8-NBD-cGMP. Since the results are consistent with full activation of Et PKG requiring cyclic nucleotide binding in all three allosteric sites, one role for the additional cGMP site may be to establish a stricter regulatory mechanism for the kinase activity than is present in the PKGs of higher organisms containing only two allosteric sites. PMID- 11914084 TI - Characterization of the membrane domain Nqo11 subunit of the proton-translocating NADH-quinone oxidoreductase of Paracoccus denitrificans. AB - The proton-translocating NADH-quinone oxidoreductase (NDH-1) of Paracoccus denitrificans consists of at least 14 unlike subunits (designated Nqo1-14). The NDH-1 is composed of two segments (the peripheral and membrane segments). The membrane domain segment appears to be made up of seven subunits (Nqo7, -8, -10 14). In this report, the characterization of the Paracoccus Nqo11 subunit has been investigated. An antibody against the C-terminal 12 amino acid residues of the Paracoccus Nqo11 subunit (Nqo11c) has been raised. The Nqo11c antibody reacted with a single band (11 kDa) of the Paracoccus membranes and cross-reacted with Rhodobactor capsulatus membranes. The Nqo11 subunit was not able to be extracted from the Paracoccus membranes by NaI or alkaline treatment, unlike the peripheral subunits (Nqo1 and Nqo6). The C-terminal region of the Paracoccus Nqo11 is exposed to the cytoplasmic phase. For further characterization of the Paracoccus Nqo11 subunit, the subunit was overexpressed in Escherichia coli by using the maltose-binding protein (MBP) fusion system. The MBP-fused Nqo11 subunit was expressed in the E. coli membranes (but not in soluble phase) and was extracted by Triton X-100. The isolated MBP-fused Nqo11 subunit interacted with the phospholipid vesicles and suppressed their membrane fluidity. Topological studies of the Nqo11 subunit expressed in E. coli membranes have been performed by using cysteine mapping and immunochemical analyses. The data suggest that the Nqo11 subunit has three transmembrane segments and its C-terminus protrudes into the cytoplasmic phase. PMID- 11914086 TI - Purification and biochemical characterization of Mycobacterium tuberculosis SuhB, an inositol monophosphatase involved in inositol biosynthesis. AB - Phosphatidylinositol is an essential component of mycobacteria, and phosphatidylinositol-based lipids such as phosphatidylinositolmannosides, lipomannan, and lipoarabinomannan are major immunomodulatory components of the Mycobacterium tuberculosis cell wall. Inositol monophosphatase (EC 3.1.3.25) is a crucial enzyme in the biosynthesis of free myo-inositol from inositol-1 phosphate, a key substrate for the phosphatidylinositol synthase in mycobacteria. Analysis of the M. tuberculosis genome suggested the presence of four M. tuberculosis gene products that exhibit an inositol monophosphatase signature. In the present report, we have focused on SuhB, which possesses the highest degree of homology with human inositol monophosphatase. SuhB gene was cloned into an E. coli expression vector to over-produce a His-tagged protein, which was purified and characterized. SuhB required divalent metal ions for functional inositol monophosphatase activity, with Mg(2+) being the strongest activator. Inositol monophosphatase activity catalyzed by SuhB was inhibited by the monovalent cation lithium (IC(50) = 0.9 mM). As anticipated, inositol-1-phosphate was the preferred substrate (K(m) = 0.177 +/- 0.025 mM; k(cat) = 3.6 +/- 0.2 s(-)(1)); however, SuhB was also able to hydrolyze a variety of polyol phosphates such as glucitol-6 phosphate, glycerol-2-phosphate, and 2'-AMP. To provide further insight into the structure-function relationship of SuhB, different mutant proteins were generated (E83D, D104N, D107N, W234L, and D235N). These mutations almost completely abrogated inositol monophosphatase activity, thus underlining the importance of these residues in inositol-1-phosphate dephosphorylation. We also identified L81 as a key residue involved in sensitivity to lithium. The L81A mutation rendered SuhB inositol monophosphatase activity 10-fold more resistant to inhibition by lithium (IC(50) = 10 mM). These studies provide the first steps in the delineation of the biosynthesis of the key metabolite inositol in M. tuberculosis. PMID- 11914087 TI - Using 2-aminopurine fluorescence to detect base unstacking in the template strand during nucleotide incorporation by the bacteriophage T4 DNA polymerase. AB - The fluorescence of the base analogue 2-aminopurine (2AP) was used to detect physical changes in the template strand during nucleotide incorporation by the bacteriophage T4 DNA polymerase. Fluorescent enzyme-DNA complexes were formed with 2AP placed in the template strand opposite the primer terminus (the n position) and placed one template position 5' to the primer terminus (the n + 1 position). The fluorescence enhancement for 2AP at the n position was shown to be due to formation of the editing complex, which indicates that the 2AP-T terminal base pair is recognized primarily as a mismatch. 2AP fluorescence at the n + 1 position, however, was a reporter for DNA interactions in the polymerase active center that induce intrastrand base unstacking. T4 DNA polymerase produced base unstacking at the n + 1 position following formation of the phosphodiester bond. Thus, the increase in fluorescence intensity for 2AP at the n + 1 position could be used to measure the nucleotide incorporation rate in primer extension reactions in which 2AP was placed initially at the n + 2 position. Primer extension occurred at the rate of about 314 s(-1). The amount of base unstacking at the template n + 1 position was sensitive to the local DNA sequence. More base unstacking was detected for DNA substrates with an A-T base pair at the primer terminus compared to C-G or G-C base pairs. Since proofreading is also increased by A-T base pairs compared to G-C base pairs at the primer terminus, we propose that base unstacking may provide an opportunity for the DNA polymerase to reexamine the primer terminus. PMID- 11914088 TI - Site-resolved energetics in DNA triple helices containing G*TA and T*CG triads. AB - Recognition of specific sites in double-helical DNA by triplex-forming oligonucleotides has been limited until recently to sites containing homopurine homopyrimidine sequences. G*TA and T*CG triads, in which TA and CG base pairs are specifically recognized by guanine or by thymine, have now extended this recognition code to DNA target sites of mixed base sequences. In the present work, we have obtained a characterization of the stabilities of G*TA and T*CG triads, and of the effects of these triads upon canonical triads, in triple helical DNA. The three DNA triplexes investigated are formed by the folding of the 31-mers d(GAAXAGGT(5)CCTYTTCT(5)CTTZTCC) with X = G, T, or C, Y = C, A, or G, and Z = C, G, or T. We have measured the exchange rates of imino protons in each triad of the three triplexes using nuclear magnetic resonance spectroscopy. The exchange rates are used to map the local free energy of structural stabilization in each triplex. The results indicate that the stability of Watson-Crick base pairs in the G*TA and T*CG triads is comparable to that of Watson-Crick base pairs in canonical triads. The presence of G*TA and T*CG triads, however, destabilizes neighboring canonical triads, two or three positions removed from the G*TA/T*CG site. Moreover, the long-range destabilizing effects induced by the T*CG triad are larger than those induced by the G*TA triad. These findings reveal the molecular basis for the lower overall stability of G*TA- and T*CG-containing triplexes. PMID- 11914089 TI - Comparative structure analysis of tyrosine and valine residues in unprocessed silk fibroin (silk I) and in the processed silk fiber (silk II) from Bombyx mori using solid-state (13)C,(15)N, and (2)H NMR. AB - The solid-state (13)C CP-MAS NMR spectra of biosynthetically labeled [(13)C(alpha)]Tyr, [(13)C(beta)]Tyr, and [(13)C(alpha)]Val silk fibroin samples of Bombyx mori, in silk I (the solid-state structure before spinning) and silk II (the solid-state structure after spinning) forms, have been examined to gain insight into the conformational preferences of the semicrystalline regions. To establish the relationship between the primary structure of B. mori silk fibroin and the "local" structure, the conformation-dependent (13)C chemical shift contour plots for Tyr C(alpha), Tyr C(beta), and Val C(alpha) carbons were generated from the atomic coordinates of high-resolution crystal structures of 40 proteins and their characteristic (13)C isotropic NMR chemical shifts. From comparison of the observed Tyr C(alpha) and Tyr C(beta) chemical shifts with those predicted by the contour plots, there is strong evidence in favor of an antiparallel beta-sheet structure of the Tyr residues in the silk fibroin fibers. On the other hand, Tyr residues take a random coil conformation in the fibroin film with a silk I form. The Val residues are likely to assume a structure similar to those of Tyr residues in silk fiber and film. Solid-state (2)H NMR measurements of [3,3-(2)H(2)]Tyr-labeled B. mori silk fibroin indicate that the local mobility of the backbone and the C(alpha)-C(beta) bond is essentially "static" in both silk I and silk II forms. The orientation-dependent (i.e., parallel and perpendicular to the magnetic field) solid-state (15)N NMR spectra of biosynthetically labeled [(15)N]Tyr and [(15)N]Val silk fibers reveal the presence of highly oriented semicrystalline regions. PMID- 11914090 TI - Interactions of the antimicrobial peptides temporins with model biomembranes. Comparison of temporins B and L. AB - Temporins are short (10-13 amino acids) and linear antimicrobial peptides first isolated from the skin of the European red frog, Rana temporaria, and are effective against Gram-positive bacteria and Candida albicans. To get insight into their mechanism(s) of action, we compared the effects on model membranes exerted by two members of this family, viz., temporin B (LLPIVGNLLKSLL-NH(2)) and temporin L (FVQWFSKFLGRIL-NH(2)). More specifically, we measured their insertion into lipid monolayers as well as their effects on the structural dynamics of liposomal bilayers as revealed by diphenylhexatriene (DPH)- and pyrene-labeled phospholipids. We also observed the impact of these peptides on the topology of giant vesicles. Both temporins readily penetrate into lipid monolayers, their intercalation being enhanced in the presence of the common bacterial negatively charged phospholipid phosphatidylglycerol. Instead, the eukaryotic lipid cholesterol did to some extent counteract their penetration into the lipid films. Both temporin B and temporin L caused an enrichment of phospholipids in the bilayers, and in the presence of 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphoglycerol (POPG), these peptides increased acyl chain order. Temporin B had practically no effect on giant liposomes composed of 1-stearoyl-2-oleoyl-sn glycero-3-phosphocholine (SOPC), whereas rapid vesiculation was observed when POPG was present. In contrast, temporin L induced vesiculation of both SOPC and SOPC/POPG giant vesicles while the presence of cholesterol in SOPC giant vesicles attenuated this effect. PMID- 11914091 TI - Pathway of ADP-stimulated ADP release and dissociation of tethered kinesin from microtubules. Implications for the extent of processivity. AB - Kinesin binds to microtubules with half-site ADP release to form a tethered intermediate with one attached head without nucleotide and one tethered head that retains its bound ADP. For DKH405 containing amino acid residues 1-405 of Drosophila kinesin, release of the remaining ADP from the tethered head is slow (0.05 s(-1)), but release is accelerated by added ADP or ATP. The maximum rate of ADP-stimulated dissociation of tethered DKH405 from the microtubule is approximately 12 s(-1) as determined by turbidity. Parallel measurements of ADP stimulated release of 2'(3')-O-(N-methylanthraniloyl)-ADP (mantADP) from the tethered intermediate by fluorescence indicate that the reaction is biphasic with a fast phase that occurs at a rate that is similar to dissociation. The rate of the slow phase is dependent on the concentrations of salt and microtubules and is equal in each case to the rate for bimolecular stimulation of ADP release by microtubules as measured independently. These results are consistent with a scheme in which the fast phase, with approximately one-third of the total amplitude change, is due to ADP-stimulated release of mantADP from the tethered intermediate at approximately 6 s(-1). This direct release of mantADP continues until terminated by dissociation of DKH405 from the microtubule at approximately 12 s(-1). The majority of the amplitude change thus occurs through bimolecular recombination of DKH405.mantADP with microtubules following initial dissociation. Analysis of a simple scheme indicates that hydrolysis of ATP at the attached head before the tethered head can release its ADP and become tightly bound may be the principal limitation to processivity. PMID- 11914092 TI - 4-Methyl-7-thioumbelliferyl-beta-D-cellobioside: a fluorescent, nonhydrolyzable substrate analogue for cellulases. AB - The kinetics of cellulose binding and hydrolysis by cellulases is not well understood except at steady-state conditions. For use in studies of cellulase pre steady-state and steady-state kinetics, we have prepared 4-methyl-7 thioumbelliferyl-beta-D-cellobioside (MUS-CB), a ground-state nonhydrolyzable analogue of the fluorescent cellulase substrate 4-methylumbelliferyl-beta-D cellobioside (MU-CB). MUS-CB is not hydrolyzed by the catalytic domain of cellulase E1 from Acidothermus cellulolyticus under conditions where this enzyme rapidly degrades MU-CB. Thermodynamic parameters describing the steady-state binding of MUS-CB to Thermobifida fusca cellulase Cel6A are similar to those for MU-CB, indicating that MUS-CB can be used in place of MU-CB to study binding events in the Cel6A active-site cleft. In the pre-steady-state, MUS-CB binds to Cel6A by a simple, one-step bimolecular association reaction. It is anticipated that similar thio-containing 4-methylumbelliferyl compounds will have applications in studies of other enzyme systems. PMID- 11914093 TI - Alteration of substrate selectivity through mutation of two arginine residues in the binding site of amadoriase II from Aspergillus sp. AB - Amadoriases I and II are deglycation isoenzymes from Aspergillus sp. of potential relevance for treatment of diabetic complications resulting from excessive protein glycation. Amadoriase II has a preference for anionic substrate with a K(m) of 0.23 and 2.53 mM for fructosylglycine and fructosylpropylamine, respectively. In contrast, the corresponding K(m) values for amadoriase I are 9.75 and 0.023 mM, respectively. Chemical modification of amadoriase II with p hydroxyphenylglyoxal, a specific arginine-modifying reagent, resulted in an inhibition of enzyme activity toward fructosylglycine, while having less effect on the enzymatic activity toward fructosylpropylamine. Peptide mapping and subsequent mass spectrometry analysis suggest that Arg(112) is one of the sites of p-hydroxyphenylglyoxal modification. Sequence alignment between amadoriase I and amadoriase II revealed that two glutamic acids in amadoriase I align to Arg(112) and Arg(114) in amadoriase II. Site-directed mutation of amadoriase II (R112E, R114E) resulted in reversal of the enzymatic activities toward fructosylglycine and fructosylpropylamine. Our results suggested that Arg(112) and Arg(114) are responsible for the high affinity of amadoriase II toward anionic substrates and determine the substrate selectivity of the enzyme. PMID- 11914094 TI - Kinetic characterization of yeast pyruvate carboxylase isozyme pyc1. AB - Yeast (Saccharomyces cerevisiae) is unusual in being the only organism thus far identified as having two genes for pyruvate carboxylase. The expression of the two isozymes Pyc1 and Pyc2 appears to be differentially regulated, and since both are expressed cytoplasmically, this suggests that they have different properties. To the present, little has been done to characterize these isozymes, and almost all of the published kinetic information on yeast pyruvate carboxylase comes from measurements of enzyme prepared from bakers' yeast which is likely to be a mixture of both isozymes. Here we have measured basic kinetic parameters for Pyc1 and found that the K(a) of this isozyme for acetyl CoA is in the order of 8-10 fold higher than previously recorded, suggesting that Pyc1 and Pyc2 may be differentially regulated by this effector. Pyc1 is highly dependent on the presence of acetyl CoA for activity and in this respect is similar to chicken liver pyruvate carboxylase. However, unlike the chicken liver enzyme, the quaternary structure of the enzyme is quite stable in the absence of acetyl CoA, and the major locus of action of this effector appears to lie outside of the stimulation of the biotin carboxylation reaction. PMID- 11914095 TI - Effects of benzo[a]pyrene adduct stereochemistry on downstream DNA replication in vitro: evidence for different adduct conformations within the active site of DNA polymerase I (Klenow fragment). AB - The presence of bulky adducts in DNA is known to interfere with DNA replication not only at the site of the lesion but also at positions up to 5 nucleotides past the adduct location. Kinetic studies of primer extension by exonuclease-deficient E. coli DNA polymerase I (Klenow fragment) (KF) when (+)-trans- or (+)-cis-B[a]P N(2)-dG adducts were positioned in the double-stranded region of the primer templates showed that both stereoisomers significantly block downstream replication. However the (+)-cis adduct, which causes a stronger inhibition of the nucleotides insertion across from and immediately past the lesion, affected the downstream replication to a much smaller extent than did the (+)-trans adduct, especially when the B[a]P-modified dG was properly paired with a dC. The effects of mismatches across from the adduct and the sequence context surrounding the adduct were also dependent on the stereochemistry of the B[a]P adduct. Thus, the identity of the nucleotide across from the adduct that provided the best downstream replication was different for the (+)-cis and (+)-trans adducts, a factor that might differentially contribute to the mutagenic bypass of these lesions. These findings provide strong direct evidence that the conformations of the (+)-cis and (+)-trans adducts within the active site of KF are significantly different and probably differentially affect the interactions of the polymerase with the minor groove, thereby leading to different replication trends. The stereochemistry of the adduct was also found to differentially affect the sequence-mediated primer-template misalignments, resulting in different consequences during the bypass of the lesion. PMID- 11914096 TI - A stable dimer in the pH-induced equilibrium unfolding of the homo-hexameric enzyme 4-oxalocrotonate tautomerase (4-OT). AB - 4-Oxalocrotonate tautomerase (4-OT) is a multimeric, bacterial enzyme comprised of 6 identical 62-amino acid subunits, which associate under native conditions to form a homo-hexameric structure stabilized entirely by noncovalent interactions. We have previously shown that the GuHCl-induced equilibrium unfolding of 4-OT at pH 8.5 is well modeled as a two-state process involving only hexamer and unfolded monomer; and we have obtained spectroscopic evidence that intermediate state(s) is (are) populated in the equilibrium unfolding reaction at pHs 6.0 and 7.4 [Silinski, P., Allingham, M. J., and Fitzgerald, M. C. (2001) Biochemistry 40, 4493-4502]. Here, we report on the pH-induced equilibrium unfolding of 4-OT using size-exclusion chromatography (SEC), far-UV-circular dichroism (CD) spectroscopy, and catalytic activity measurements over the pH range from 1.5 to 10.1. Our results indicate that the native hexamer of 4-OT is the predominant species in solution at pHs > or =6.2, that a partially folded dimeric state of 4-OT is stabilized in solution at pH 4.8, and that the enzyme is largely denatured in strongly acidic solutions (pH < or =3.1). GuHCl-induced equilibrium unfolding studies on 4-OT at pH 4.8 indicate that the folded 4-OT dimer populated at this pH is stabilized by 11.7 kcal.mol(-1). The results of biophysical studies on a fluorescent analogue of the enzyme, 4-OT(F50Y), and the results of UV photo-cross linking studies on a synthetically derived 4-OT analogue, 4-OT(P1Bpa), suggest the polypeptide chains in the 4-OT dimer are nativelike in structure with the exception of their C-termini. PMID- 11914097 TI - Mechanistic analyses of catalysis in human pancreatic alpha-amylase: detailed kinetic and structural studies of mutants of three conserved carboxylic acids. AB - The roles of three conserved active site carboxylic acids (D197, E233, and D300) in the catalytic mechanism of human pancreatic alpha-amylase (HPA) were studied by utilizing site-directed mutagenesis in combination with structural and kinetic analyses of the resultant enzymes. All three residues were mutated to both alanine and the respective amide, and a double alanine mutant (E233A/D300A) was also generated. Structural analyses demonstrated that there were no significant differences in global fold for the mutant enzymes. Kinetic analyses were performed on the mutants, utilizing a range of substrates. All results suggested that D197 was the nucleophile, as virtually all activity (>10(5)-fold decrease in k(cat) values) was lost for the enzymes mutated at this position when assayed with several substrates. The significantly greater second-order rate constant of E233 mutants on "activated" substrates (k(cat)/K(m) value for alpha-maltotriosyl fluoride = 15 s(-)(1) mM(-)(1)) compared with "unactivated" substrates (k(cat)/K(m) value for maltopentaose = 0.0030 s(-)(1) mM(-)(1)) strongly suggested that E233 is the general acid catalyst, as did the pH-activity profiles. Transglycosylation was favored over hydrolysis for the reactions of several of the enzymes mutated at D300. At the least, this suggests an overall impairment of the catalytic mechanism where the reaction then proceeds using the better acceptor (oligosaccharide instead of water). This may also suggest that D300 plays a crucial role in enzymic interactions with the nucleophilic water during the hydrolysis of the glycosidic bond. PMID- 11914099 TI - Evidence for myocardial synthesis of aldosterone producing myocardial fibrosis in man. PMID- 11914098 TI - Aldosterone synthase (CYP11B2) expression and myocardial fibrosis in the failing human heart. AB - The pathway of tissue aldosterone production may exist in the heart, and may be an important contributory factor to myocardial fibrosis and cardiac remodelling in the failing heart. CYP11B2 (aldosterone synthase) catalyses the final step of aldosterone production. The aim of the present study was to determine whether CYP11B2 and CYP11B1 (11beta-hydroxylase) are expressed in myocardial tissues, and whether these enzymes contribute to collagen accumulation and myocardial dysfunction in the failing human heart. Endomyocardial tissues were obtained from 23 patients with chronic heart failure (CHF) and 10 controls. CYP11B2 and CYP11B1 mRNA levels were measured by real-time quantitative reverse transcriptase-PCR. The myocardial collagen volume fraction (CVF) was determined by digital planimetry. CYP11B2 mRNA expression was greater in the CHF group than in the controls (P<0.05), while CYP11B1 mRNA was barely expressed in either group. There was a positive correlation between CYP11B2 mRNA levels and CVF (r=0.64, P=0.001). CYP11B2 mRNA was particularly highly expressed in subgroups of CHF patients with a large left ventricular end-systolic diameter (>55 mm) or a low left ventricular ejection fraction (<30%). CYP11B2 mRNA expression and CVF were lower in a CHF subgroup treated with a combination of spironolactone and angiotensin-converting enzyme inhibitors (ACEIs) than in a subgroup not treated with these drugs. In conclusion, this study has shown that increased myocardial expression of CYP11B2 mRNA is associated with increased myocardial fibrosis and with the severity of left ventricular dysfunction in human CHF. In addition, CYP11B2 expression and cardiac fibrosis are found to be decreased in CHF patients on drug therapy comprising spironolactone combined with ACEIs. PMID- 11914100 TI - Liver electrolysis: pH can reliably monitor the extent of hepatic ablation in pigs. AB - Electrolysis is a method of tissue ablation that creates chemical species and a pH gradient in response to direct current. Initial studies of electrolysis in animal models and humans have shown that it is a safe, predictable and effective process for destroying normal and tumour-bearing liver in a linear, dose dependent manner. Presently, the amount of current that is applied (in coulombs) has to be calculated using historical data, with inherent inaccuracy. The present study tested whether pH could be used as a real-time monitor in order to predict more accurately the extent of necrosis. A total of 70 electrolytic lesions were created in 14 pigs, with pH monitoring of the lesion edge. The normal range of pH values was 6.5-8.7. A pH of less than 6 (at the anode) or more than 9 (at the cathode) reflected total cellular necrosis. When a pH value was recorded between 6.0 and 6.5 at the anode or between 8.7 and 9.0 at the cathode, the presence of necrosis was variable. In conclusion, during electrolytic ablation, pH measurement can monitor the extent of the induced necrosis. PMID- 11914101 TI - Nitric oxide and cardiac parasympathetic control in human heart failure. AB - Cardiac parasympathetic control has prognostic significance in heart failure, but the control mechanisms of this system remain poorly defined. We have demonstrated previously a facilitatory role for nitric oxide (NO) in the parasympathetic control of heart rate in young healthy human subjects. In view of the complex abnormalities of regional NO activity observed in chronic heart failure, we now aim to establish if this mechanism is active in subjects with this condition. Groups of 12 heart failure patients [NYHA class II-III; mean age 52 years (range 38-67 years)] and 12 age/sex-matched healthy control subjects [mean age 50 years (range 36-62 years)] were studied. Heart rate variability and baroreflex sensitivity were measured during inhibition of endogenous NO production with N(G) monomethyl-l-arginine (l-NMMA; 3 mg.h(-1).kg(-1)) and during administration of an equipressor dose of the control vasoconstrictor phenylephrine (12-36 microg.h( 1).kg(-1)). Basal levels of nitrate+nitrite were measured in the plasma as an indication of systemic NO production. In the heart failure patients, despite an equal rise in blood pressure with both drugs, high-frequency indices of heart rate variability increased less with l-NMMA than with phenylephrine: RMSSD (root mean square of successive RR-interval differences) increased by 4+/-2 compared with 26+/-8 ms (P<0.001) and high-frequency power increased by 97+/-62 compared with 1372+/-861 ms(2) (P<0.001). The increases in cross-spectral baroreflex sensitivity were also lower with l-NMMA than with phenylephrine [high-frequency alpha-index, 2.2+/-1.3 and 12.6+/-3.8 ms/mmHg respectively (P<0.001); low frequency alpha-index, 1.3+/-0.9 and 4.3+/-1.7 ms/mmHg respectively (P<0.05)]. Healthy subjects showed a similar discrepancy in the response of high-frequency indices of heart rate variability to the two drugs, although baroreflex sensitivity responses were significantly different only for the high-frequency alpha-index. Levels of plasma nitrate+nitrite were significantly higher in the heart failure patients compared with controls. These data demonstrate that baroreflex-mediated cardiac parasympathetic activation in human heart failure, as in health, is dependent upon endogenous NO synthesis. PMID- 11914102 TI - Fatty acids and cytokine mRNA expression in human osteoblastic cells: a specific effect of arachidonic acid. AB - Epidemiological, clinical and experimental evidence suggests that fatty acids have a modulatory effect on bone metabolism in animals and humans. To investigate this hypothesis, we evaluated the effects of three different fatty acids, arachidonic acid (AA), eicosapentaenoic acid (EPA) and oleic acid (OA), on the expression of cytokines involved in bone remodelling. Cytokine mRNAs in the human osteoblast-like cell line MG-63 were quantified by reverse transcription-PCR. AA induced increased expression of interleukin-1alpha, interleukin-1beta, tumour necrosis factor-alpha and macrophage colony-stimulating factor mRNAs in a time- and dose-dependent manner. EPA and OA had no stimulatory effects, but instead caused a significant inhibition of AA-induced cytokine mRNA expression. Cell treatment with calphostin C, an inhibitor of protein kinase C (PKC), and cellular PKC down-regulation experiments independently resulted in significant inhibition of AA-induced cytokine expression, suggesting that a PKC-dependent mechanism accounts for the effects of AA on cytokine production. In conclusion, our study demonstrates specific effects of fatty acids on cytokine gene expression in human osteoblast-like cells. The clinical relevance of our findings requires further investigation. PMID- 11914103 TI - Non-competitive immunochemiluminometric assay for cardiotrophin-1 detects elevated plasma levels in human heart failure. AB - Cardiotrophin-1 (CT-1) leads to a specific form of ventricular hypertrophy characterized by sarcomeres added in series, and has been reported to be elevated in heart failure. Previous competitive assays for CT-1 necessitate the extraction of plasma and involve prolonged incubations. We describe the development of a non competitive assay for CT-1 that can measure plasma levels without the need for extraction. Two antibodies specific for the mid-section (amino acids 105-120) and C-terminal (amino acids 186-199) portions of CT-1 were developed in rabbits. One antibody was immobilized and used as the capture antibody. The other antibody was affinity purified and biotinylated. Unextracted plasma was incubated with these antibodies, and detection was with methylacridinium ester-labelled streptavidin. Plasma was obtained from 40 patients with heart failure and 40 normal control subjects. The non-competitive assay demonstrated a linear increase in chemiluminescence (measured as relative light units) with increasing amounts of full-length recombinant CT-1, with no evidence of a hook effect at high concentrations. The lower limit of detection was 2.9 fmol/ml. Intra-assay coefficients of variation ranged from 3.1% to 4.2% in the 10-40 fmol/well concentration range, and interassay coefficients of variation ranged from 3.5% to 4.5% in the 550-950 fmol/ml range. Measurements of CT-1 levels in patients with heart failure (median 166.5 fmol/ml; range 49.5-2788 fmol/ml) revealed very significantly elevated levels compared with those in normal controls (median 43.5 fmol/ml; range 11.2-258.6 fmol/ml; P<0.0001 by Mann-Whitney test). At a CT-1 concentration of 68 fmol/ml, sensitivity and specificity were 95% and 82.5% respectively. Thus this new non-competitive immunochemiluminometric assay for CT 1 could successfully detect full-length recombinant CT-1 in unextracted plasma, and demonstrated that plasma levels of CT-1 were significantly elevated in patients with heart failure. PMID- 11914104 TI - Extracellular matrix regulation of drug resistance in small-cell lung cancer. AB - Tumour recurrence following chemotherapy remains a major obstacle to the cure of many cancers. This is exemplified by small-cell lung cancer (SCLC). Host-tumour interactions are central to tumour survival and proliferation. We hypothesized that a factor(s) within the local environment of SCLC cells could provide a survival signal or block a death signal, thereby accounting for the protection of SCLC cells from chemotherapy-induced apoptosis. Here we review recent work undertaken in our laboratory addressing this issue. We have shown that, in vivo, SCLC cells are surrounded by an extensive stroma of extracellular matrix (ECM) at both primary and metastatic sites which contains, among other proteins, fibronectin, laminin and collagen IV. Furthermore, adhesion of SCLC cells to fibronectin, laminin and collagen IV through beta1 integrins enhances tumorigenicity and confers resistance to apoptosis induced by standard chemotherapeutic agents, including etoposide, cis-platinum and adriamycin. Adhesion to ECM proteins stimulated protein tyrosine kinase (PTK) activity in both untreated and etoposide-treated cells. This effect could be completely blocked by a selective PTK inhibitor or by a function-blocking beta1 integrin antibody. PTK activation was found to block chemotherapy-induced activation of the death protease caspase-3 and, hence, apoptosis. Adhesion to ECM or treatment with a PTK inhibitor did not affect etoposide inhibition of topoisomerase II. Thus adhesion to ECM through beta1 integrins protects SCLC cells from chemotherapy-induced caspase-3 activation and apoptosis by activating PTK signalling downstream of DNA damage. Survival of tumour cells attached to ECM within this microenvironment could explain the local recurrence of SCLC and other tumours that is often seen clinically after chemotherapy. PMID- 11914105 TI - Effect of prothrombin and its activation fragments on calcium oxalate crystal growth and aggregation in undiluted human urine in vitro: relationship between protein structure and inhibitory activity. AB - In recent years there has been great interest in the putative role of prothrombin and its activation peptides, especially the urinary form of prothrombin fragment 1, in the pathogenesis of calcium oxalate (CaOx) urolithiasis. Previously, we showed that prothrombin and its activation peptides inhibit CaOx crystallization in inorganic conditions in vitro. The aim of the present study was to determine if this inhibitory activity is retained in undiluted human urine and, therefore, whether it is likely to have any influence under physiological conditions. A secondary objective was to assess the relationship between the structures of the proteins and their inhibitory activities. Prothrombin was purified from Prothrombinex-HT, cleaved with thrombin and the resulting fragment 1 (F1) and fragment 2 (F2) were purified. The purity of each protein was confirmed by SDS/PAGE, and their effects on CaOx crystallization in undiluted ultrafiltered human urine were determined at a final concentration 80.65 nmol/l using Coulter Counter and [(14)C]oxalate analysis. The precipitated crystals were visualized using scanning electron microscopy. The Coulter Counter data revealed that, whereas prothrombin and its activation peptides did not affect the urinary metastable limit and the size of the precipitated particles, F1 did significantly reduce the latter. These findings were corroborated with scanning electron microscopy which also revealed that the reduction in particle size caused by F1 resulted from a decrease in the degree of crystal aggregation, rather than in the size of the individual crystals. The [(14)C]oxalate data showed that none of the proteins added significantly inhibited the mineral deposition. It was concluded that with the exception of F1, which does inhibit CaOx crystal aggregation, prothrombin and its activation peptides do not alter the deposition and aggregation of CaOx crystals in ultrafiltered human urine in vitro. Also, the gamma-carboxyglutamic acid domain of prothrombin and F1, which is absent from thrombin and F2, is the region of the molecules that determines their potent inhibitory effects. The superior potency of F1, compared with prothrombin, probably results from the molecule's greater charge-to-mass ratio. PMID- 11914106 TI - Delayed cardioprotective effects of exercise in dogs are aminoguanidine sensitive: possible involvement of nitric oxide. AB - Dogs were subjected to exercise on a treadmill, using a protocol in which the speed and slope were increased every 3 min, and which elevated both heart rate (to a mean of 198+/-14 beats.min(-1)) and mean arterial blood pressure (to 150+/ 4 mmHg). Then, 24 or 48 h later, the dogs were anaesthetized with a mixture of alpha-chloralose and urethane and subjected to a 25 min occlusion of the left anterior descending coronary artery. The control dogs (instrumented but not exercised) were subjected to the same procedure. In some dogs the nitric oxide synthase inhibitor aminoguanidine (50 mg.kg(-1); intravenous) was administered 30 min before occlusion. Baroreflex sensitivity (BRS) was determined by the rapid bolus injection of phenylephrine 60 min before, and again 3 min after, the onset of occlusion. Exercise markedly reduced the consequences of coronary artery occlusion 24 h (but not 48 h) later, without modifying myocardial tissue blood flow. In the exercised dogs there were reductions in arrhythmia severity [ventricular fibrillation (VF) during occlusion, 0%; survival from the combined ischaemia/reperfusion insult, 70%] compared with controls (VF during occlusion, 36%; survival, 9%). BRS was preserved during occlusion in the exercised dogs (before occlusion, 1.60+/-0.54 ms.mmHg(-1); 3 min after occlusion, 1.37+/-0.4 ms.mmHg(-1)), but not in controls (before occlusion, 1.28+/-0.29 ms.mmHg(-1); 3 min after occlusion, 0.45+/-0.12 ms.mmHg(-1); P<0.05), and other ischaemic changes (inhomogeneity of electrical activation and changes in the ST-segment, recorded over the ischaemic region) were also less marked in the exercised dogs. Exercise-induced cardioprotection was abolished by aminoguanidine (VF during occlusion, 25%; survival, 0%). The results show that even a single period of exercise affords delayed protection against ischaemia/reperfusion-induced VF and other ischaemic changes. Since this protection is abolished by aminoguanidine, and since (inducible) NO synthase activity was elevated 3-fold in left ventricular samples 24 h after exercise, we suggest that this protection is mediated by nitric oxide. PMID- 11914107 TI - A European multicentre, placebo-controlled supplementation study with alpha tocopherol, carotene-rich palm oil, lutein or lycopene: analysis of serum responses. AB - Increased levels of oxidative stress have been implicated in tissue damage and the development of chronic diseases, and dietary antioxidants may reduce the risk of oxidative tissue damage. As part of a European multicentre project, several studies were undertaken with the aim of testing whether the consumption of foods rich in carotenoids reduces oxidative damage to human tissue components. We describe here the serum response of carotenoids and tocopherols upon supplementation with carotenoids from natural extracts (alpha-carotene+beta carotene, lutein or lycopene; 15 mg/day) and/or with alpha-tocopherol (100 mg/day) in a multicentre, placebo-controlled intervention study in 400 healthy male and female volunteers, aged 25-45 years, from five European regions (France, Northern Ireland, Republic of Ireland, The Netherlands and Spain). Supplementation with alpha-tocopherol increased serum alpha-tocopherol levels, while producing a marked decrease in serum gamma-tocopherol. Supplementation with alpha- + beta-carotene (carotene-rich palm oil) resulted in 14-fold and 5-fold increases respectively in serum levels of these carotenoids. Supplementation with lutein (from marigold extracts) elevated serum lutein (approx. 5-fold), zeaxanthin (approx. doubled) and ketocarotenoids (although these were not present in the supplement), whereas lycopene supplementation (from tomato paste) resulted in a 2-fold increase in serum lycopene. The isomer distributions of beta-carotene and lycopene in serum remained constant regardless of the isomer composition in the capsules. In Spanish volunteers, additional data showed that the serum response to carotenoid supplementation reached a plateau after 4 weeks, and no significant side effects (except carotenodermia) or changes in biochemical or haematological indices were observed throughout the study. This part of the study describes dose-time responses, isomer distribution, subject variability and side effects during supplementation with the major dietary carotenoids in healthy subjects. PMID- 11914108 TI - Glucose flux is normalized by compensatory hyperinsulinaemia in growth hormone induced insulin resistance in healthy subjects, while skeletal muscle protein synthesis remains unchanged. AB - The aim of this present investigation was to study the relationship between the reduction in insulin sensitivity accompanying 5 days of treatment with growth hormone (GH; 0.05 mg.24 h(-1).kg(-1)) and intracellular substrate oxidation rates in six healthy subjects, while maintaining glucose flux by a constant glucose infusion and adjusting insulin infusion rates to achieve normoglycaemia (feedback clamp). Protein synthesis rates in skeletal muscle (flooding dose of L [(2)H(5)]phenylalanine) were determined under these conditions. We also compared changes in insulin sensitivity after GH treatment with simultaneous changes in energy requirements, protein synthesis rates, nitrogen balance, 3-methylhistidine excretion in urine, body composition and the hormonal milieu. After GH treatment, 70% more insulin was required to maintain normoglycaemia (P<0.01). The ratio between glucose infusion rate and serum insulin levels decreased by 34% at the two levels of glucose infusion tested (P<0.05). Basal levels of C-peptide, insulin-like growth factor (IGF)-I and IGF-binding protein-3 increased almost 2 fold, while levels of glucose, insulin, glucagon, GH and IGF-binding protein-1 remained unchanged. Non-esterified fatty acid levels decreased (P<0.05). In addition, 24 h urinary nitrogen excretion decreased by 26% (P<0.01) after GH treatment, while skeletal muscle protein synthesis and 3-methylhistidine excretion in urine remained unchanged. Energy expenditure increased by 5% (P<0.05) after treatment, whereas fat and carbohydrate oxidation were unaltered. In conclusion, when glucose flux was normalized by compensatory hyperinsulinaemia under conditions of GH-induced insulin resistance, intracellular rates of oxidation of glucose and fat remained unchanged. The nitrogen retention accompanying GH treatment seems to be due largely to improved nitrogen balance in non-muscle tissue. PMID- 11914109 TI - Advanced analysis of spontaneous baroreflex sensitivity, blood pressure and heart rate variability in patients with dilated cardiomyopathy. AB - Baroreflex sensitivity (BRS) is an important parameter in the classification of patients with reduced left ventricular function. This study aimed at investigating BRS in patients with dilated cardiomyopathy (DCM) and in healthy subjects (controls), as well as comparing the values of BRS parameters with parameters of heart rate variability (HRV) and blood pressure variability (BPV). ECG, continuous blood pressure and respiration curves were recorded for 30 min in 27 DCM patients and 27 control subjects. The Dual Sequence Method (DSM) includes the analysis of spontaneous fluctuations in systolic blood pressure and the corresponding beat-to-beat intervals of heart rate to estimate bradycardic, opposite tachycardic and delayed baroreflex fluctuations. The number of systolic blood pressure/beat-to-beat interval fluctuations in DCM patients was reduced in comparison with controls (DCM patients: male, 154.4+/-93.9 ms/mmHg; female, 93.7+/-40.5 ms/mmHg; controls: male, 245.5+/-112.9 ms/mmHg; female, 150.6+/-55.8 ms/mmHg, P<0.05). The average slope in DCM patients was lower than in controls (DCM, 5.3+/-1.9 ms/mmHg; controls, 8.0+/-5.4 ms/mmHg; P<0.05). Discriminant function analysis showed that, in the synchronous range of the standard sequence method, the DCM and control groups could be discriminated to only 76% accuracy, whereas the DSM gave an improved accuracy of 84%. The combination of six parameters of HRV, BPV and DSM gives an accuracy of classification of 96%, whereas six parameters of HRV and BPV could separate the two groups to only 88% accuracy. Thus the DSM leads to an improved characterization of autonomous regulation in order to differentiate between DCM patients and healthy subjects. BRS in DCM patients is significantly reduced and apparently less effective. PMID- 11914110 TI - REDOX regulation of early embryo development. AB - Preimplantation embryonic development is associated with a change in preference in energy metabolism pathways. Although oxidative phosphorylation is obligatory in most species throughout preimplantation development, an increasing role for energy derived from glycolysis is associated with compaction and blastulation. Such a shift in metabolic pathway preference is desirable as the embryo faces an increasingly hypoxic environment in utero. We hypothesize that this shift in metabolic preference is associated with a change in the reduction-oxidation (REDOX) state within the embryo, affecting not only the energy production required for development, but also the activity of REDOX-sensitive transcription factors, which may alter gene expression patterns. Shifts in intracellular REDOX state may also contribute to spatial differences in cell activity, especially after compaction, and perhaps even major embryonic events such as fertilization, genome activation and cellular differentiation. PMID- 11914111 TI - Placental peptides as markers of gestational disease. AB - The human placenta produces a wide range of important peptides, of which an intricate balance is required throughout pregnancy. In a gestational disease, this balance may be disturbed and the identification of such changes may be used to detect a particular pathology or to ascertain its severity. This review considers the role and association of various placental peptide markers associated with the major gestational diseases including intrauterine growth retardation, pre-term labour, pre-eclampsia, chromosomal disorders, gestational diabetes and trophoblastic disease. Potential markers that may prove more reliable and specific in their diagnostic value and that may be used for identifying patients at risk are also discussed. The importance of the new fields of genomics and proteomics in the future discovery of new peptide markers is illustrated. PMID- 11914112 TI - Behavioural significance of prolactin signalling in the central nervous system during pregnancy and lactation. AB - The role of prolactin in the regulation of mammary gland development and function during pregnancy and lactation is well established. However, in addition, prolactin appears to have a much wider role in the physiology of lactation. There is widespread expression of prolactin receptors in the hypothalamus during lactation, indicative of a multi-faceted role for prolactin in regulating hypothalamic function. During pregnancy and lactation, the maternal brain undergoes structural and functional modification, allowing the establishment of appropriate behaviour to feed and nurture the offspring, to adjust to the nutritional and metabolic demands of milk production, and to maintain appropriate hormone secretion to allow milk synthesis, secretion and ejection. The coordination of such a range of neurobiological and neuroendocrine adaptations requires an endocrine signalling mechanism, capable of communicating the reproductive state to the brain. Evidence indicates that prolactin is part of this mechanism. PMID- 11914113 TI - Embryo development and establishment of pregnancy after embryo transfer in pigs: coping with limitations in the availability of viable embryos. AB - Embryo transfer and pregnancy maintenance strategies in pigs were evaluated with reference to situations in which limited numbers of viable embryos or micromanipulated embryos are available, such as pig cloning. Development of embryos with compromised zona pellucida was compared with development of embryos with intact zona pellucida. Micromanipulation had no effect on blastocyst production rates after development in vivo or in vitro, but development in vivo improved the number of embryos reaching the blastocyst stage. Transfer of embryos with compromised zona pellucida resulted in live piglets. Several hormone treatments to maintain pregnancy were tested in a model in which three embryos were transferred into unmated recipient gilts, compared with transfer of three embryos into mated recipients. None of the hormonal treatments resulted in pregnancy rates of more than 25% at term and no more than 9% of transferred embryos survived, in comparison with 50% of the mated recipients successfully carrying 25% of transferred embryos. Lastly, the developmental potential of parthenogenetic embryos was assessed and 62% of transferred embryos resulted in pregnancies, none of which continued beyond day 55 of gestation. After co transfer of three fertilized embryos with 55-60 parthenogenetic embryos into each of six recipients, two live piglets were delivered. The results from the present study indicate that transfer of zona pellucida compromised embryos can yield litters of normal piglets. In addition, it was demonstrated in a model system involving the transfer of three fertilized embryos into mature gilts that hormonal pregnancy maintenance strategies support a low proportion of embryos to term. Lastly, the present study shows for the first time a comparably effective but novel alternative for pregnancy maintenance in the pig involving the co transfer of parthenote embryos. PMID- 11914114 TI - A possible extratubular origin of epididymal basal cells in mice. AB - The origin of basal cells in mouse epididymis was examined by counting the numbers of basal cells, intratubular mitotic figures and peritubular cells during development of the epididymis. Putative precursors of basal cells were labelled with bromodeoxyuridine and the nuclei of daughter cells were examined. Histochemical localization of cytokines was performed to gauge their involvement in migration of basal cell precursors from extratubular sources. The results indicate that basal cells may arise from extratubular sources as: (i) there was a decrease in the number of mitotic figures as the number of basal cells increased; (ii) no mitotic figures were observed in the base of the epithelium; (iii) the increase in the number of peritubular cells did not parallel the number of basal cells in all epididymal regions; (iv) division of epithelial cells into daughter cells was circumferential and not radial; (v) bromodeoxyuridine-labelled basal cell nuclei were mostly not found in the vicinity of labelled principal cell nuclei and vice versa; and (vi) the percentage of labelled basal cell nuclei was higher than that of the other cells, which is indicative of their arrival from a more highly labelled pool. In addition, no age-dependent correlation was noted between any of the cytokines tested and appearance of basal cells in the epithelium, and basal cells expressed intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1), which provides further evidence of a relationship between basal cells and immunocytes. If basal cells have an immunological function, failure of their recruitment into the epididymal epithelium at about the time of puberty may have repercussions for immunological protection of spermatozoa and, ultimately, for fertility in the adult. PMID- 11914115 TI - Increased ovulation rate in gilts treated with dihydrotestosterone. AB - Treatment with testosterone increases ovulation rate in pigs. The present study was conducted to examine the effects of 5alpha-dihydrotestosterone (DHT), a non aromatizable androgen receptor ligand, on ovulation rate and amounts of androgen receptor and FSH receptor mRNAs in postpubertal gilts. In Expt 1, ovulation rate in response to daily i.m. injections of 0, 6, 60 or 600 microg DHT kg(-1) body weight from day 13 of the oestrous cycle (day 0 = day 1 of oestrus) to the following oestrus increased with each dose of DHT (P < 0.05). The mean increase in number of corpora lutea ranged from approximately three to 17 over the three dosages of DHT. In Expt 2, gilts treated daily with 60 microg DHT kg(-1) body weight during the early follicular phase (from day 13 to day 16), coincident with follicular recruitment, or the late follicular phase (day 17 to oestrus), had higher (P < 0.05) rates of ovulation compared with gilts that received vehicle, and were not different from gilts treated with DHT from day 13 to oestrus. Percentage recovery of day 3 embryos was not altered when gilts were treated from day 13 to day 16 or from day 17 to oestrus; however, treatment of gilts with DHT from day 13 to oestrus decreased recovery of day 3 (Expt 1) or day 11 (Expt 2) conceptuses. Daily administration of 6 microg DHT kg(-1) body weight to gilts from day 13 of the oestrous cycle to the following oestrus (Expt 3) did not affect the relative amounts of androgen receptor mRNA, but increased (P < 0.05) the amounts of FSH receptor mRNA in preovulatory follicles as determined by RT PCR. The results of these experiments indicate that androgens may regulate ovulation rate in gilts. One of the roles of androgens might be regulation of the amounts of FSH receptor mRNA in ovarian follicles. PMID- 11914116 TI - Plasma FSH, inhibin A and inhibin isoforms containing pro- and -alphaC during winter anoestrus, spring transition and the breeding season in mares. AB - Ten mares were studied from February (winter anoestrus) to their second ovulation in the breeding season to investigate the relationship between resumption of ovarian cyclicity in the spring and circulating concentrations of FSH, inhibin A and inhibin isoforms containing pro- and -alphaC immunoreactivity. An additional four mares were studied during one oestrous cycle. Growth and regression of ovarian follicles were monitored by transrectal ultrasonography. The frequency of blood sampling varied from three times a week to once a day, depending on the follicular activity present. Concentrations of FSH, oestradiol, inhibin A and pro and -alphaC isoforms were low during deep winter anoestrus when minimal follicular activity was present in the ovaries. During spring transition, an increase in FSH concentration preceded the emergence of each follicular wave. Concentrations of inhibins were significantly higher (P < 0.05) during growth of anovulatory follicles in spring transition than during winter anoestrus. Plasma concentrations of oestradiol and inhibin A were significantly higher (P < 0.001, P < 0.05, respectively) during the growth of preovulatory follicles than during the growth of transitional anovulatory follicles, but concentrations of inhibin pro-alphaC isoforms did not differ between the two types of follicle. During the oestrous cycle, there was a significant inverse relationship (P < 0.001) between concentrations of FSH and the inhibins. Plasma inhibin pro-alphaC isoforms, but not inhibin A, reached a peak on the day of ovulation. The results strongly indicate that FSH regulates growth of spring anovulatory and preovulatory follicles. Inhibins are likely to contribute to negative feedback on the release of FSH from the pituitary gland both during the transitional period and the breeding season in mares. PMID- 11914117 TI - The importance of oxytocin mechanisms in the control of mouse parturition. AB - The role of oxytocin in parturition in mice was investigated. Pup birth profiles, blood samples and brains were collected from parturient mice observed under red light conditions in a reversed light:dark photoperiod. Peripheral administration of an oxytocin antagonist in a dose-dependent manner delayed the birth of subsequent pups, indicating that oxytocin is required for a normal pup birth profile. Oxytocin neurones were activated during birth as shown by both increased immediate early gene ( Fos) expression in oxytocin neurones in the supraoptic nucleus and increased plasma oxytocin concentrations during birth. In addition, the nucleus of the tractus solitarius and the olfactory bulbs, sites that process inputs to oxytocin neurones, become activated during parturition. Exposure to stress during parturition halted subsequent deliveries; at this stage plasma oxytocin concentrations were not higher than those of virgin mice, and birth was restored by administration of oxytocin. Administration of beta-adrenergic antagonist (propranolol) also restored stress-delayed birth, whereas administration of ritrodrine (beta-agonist) delayed birth in non-stressed mice, indicating that adrenergic mechanisms contribute to stress-delayed births in mice. Administration of morphine (mu-opioid agonist) delayed births transiently, but naloxone (opioid antagonist) did not prevent stress-delayed birth, indicating that endogenous opioids do not appear to contribute to neuroendocrine or uterine mechanisms that promote birth in mice. Therefore, despite evidence in oxytocin knockout mice that oxytocin is not essential for parturition in this species, the results of the present study indicate that oxytocin neurone activity and secretion contribute to the birth process in normal mice. PMID- 11914118 TI - Kinetics of early in vitro development of bovine in vivo- and in vitro-derived zygotes produced and/or cultured in chemically defined or serum-containing media. AB - The kinetics of the in vitro development of early embryos from bovine zygotes derived in vitro and in vitro were compared, investigating the effect of serum during in vitro maturation and fertilization (IVM-IVF) and in culture. Zygotes were collected from superovulated heifers or produced in vitro from immature oocytes with or without serum supplementation, and cultured subsequently in defined culture medium (SOFaaci) with or without serum supplementation. Time lapse images were recorded every 0.5 h throughout the culture period. More in vivo- than in vitro-derived zygotes developed to the compact morula or blastocyst stages (87% versus 47-54%, respectively; P < 0.05). Embryo development was blocked predominantly at the second or fourth cell cycles (28 and 29%). However, blastomeres degenerated at all cleavage stages. Serum supplementation during IVM IVF resulted in abnormally sized blastomeres at first cleavage (defined serum: 20 22% versus in vivo-derived: 8%, P < 0.05). The duration of the second, third and fifth cell cycles of in vivo-derived zygotes were 1-5 h shorter compared with those of in vitro-derived zygotes cultured under similar conditions (P < 0.05). However, the kinetics of embryo development was affected by serum during IVM-IVF and culture. The first and fourth cell cycles were prolonged by 4-5 h in the absence of serum during IVM-IVF, whereas the presence of serum during culture decreased the duration of the fourth cell cycle and triggered premature blastulation. The results of this study illustrate the differences and similarities between the morphology and developmental kinetics of in vivo- and in vitro-derived zygotes, and show how serum supplementation during IVM-IVF and culture can alter these parameters. PMID- 11914119 TI - Characterization of mouse spermatogonia by transmission electron microscopy. AB - Characteristics of the various type A, intermediate (In) and B spermatogonia were determined in C57BL/6J mice using transmission electron microscopy. Spermatogonia were photographed at all stages of the cycle of the seminiferous epithelium. Over 450 images were taken. Spermatogonia could be classified into As, Apr, Aal, A1 cells, A2 cells, A3 cells, A4 cells, intermediate type and type B cells primarily on the basis of nuclear and nucleolar characteristics. The most primitive spermatogonia (As, Apr, Aal) had mottled chromatin; A1 cells contained homogeneously finely granular chromatin throughout the nucleus; A2, A3, A4 and intermediate type spermatogonia had progressively increasing amounts of chromatin encrusting the nuclear envelope; type B spermatogonia had less heterochromatin along the nuclear envelope, although the particles were more dense and rounded than in intermediate type spermatogonia. Mitochondrial size and position of Golgi complexes varied in different types of spermatogonia. These data show that types of spermatogonia can be differentiated such that these characteristics can be used in functional studies. PMID- 11914120 TI - Differential effects of glucose and fructose on hexose metabolism in dog spermatozoa. AB - Incubation of dog spermatozoa with 10 mmol l(-1) glucose or fructose rapidly increased the intracellular content of glucose 6-phosphate and fructose 6 phosphate, although the effect of fructose was greater. These effects were correlated with increases in ATP, ribose 5-phosphate and glycogen contents, and in the rates of formation of L-lactate and CO2. In all cases, except for ATP and glycogen, the effect of fructose was greater than that of glucose. The total hexokinase activity of the crude extracts of dog spermatozoa was more sensitive to fructose than to glucose at lower concentrations (0.1-3.0 mmol l(-1)). Both monosaccharides induced a fast and intense increase in the overall tyrosine phosphorylation of dog spermatozoa, although their specific induced phosphorylation patterns differed slightly. Glut 3 and Glut 5 hexose transporters were the main hexose transporters in dog spermatozoa; however, other possible SGLT family-related hexose transporters were also localized. These data indicate that, at concentrations from 1 mmol l(-1) to 10 mmol l(-1), fructose has a stronger effect than glucose on hexose metabolism of dog spermatozoa. These differences appear to be related to variations in the sensitivity of hexokinase activity. Moreover, the differential hexose metabolism induced by the two sugars had distinct effects on the function of dog spermatozoa, as revealed by the diverse patterns of tyrosine phosphorylation. PMID- 11914121 TI - Birth of pups after transfer of mouse embryos derived from vitrified preantral follicles. AB - Preantral follicles mechanically isolated from the ovaries of 12-day-old mice were exposed to 2 mol ethylene glycol l(-1) for 2 or 5 min and then to a vitrification solution containing 6 mol ethylene glycol l(-1) and 0.3 mol raffinose l(-1) for 0.5, 1.0 or 2.0 min before vitrification. The vitrified and fresh preantral follicles were treated with collagenase, and the oocyte-granulosa cell complexes (OGCs) obtained were cultured in vitro for 10 days in membrane inserts. Preantral follicles exposed to 2 mol ethylene glycol l(-1) for 5 min and then to the vitrification solution for 0.5 or 1.0 min showed the highest survival rates after warming. The follicular loss after warming was approximately 20%. After in vitro culture, the proportion of viable OGCs from the vitrified follicles was 10% lower than that of the fresh preantral follicles. There were no differences in the rates of maturation, fertilization and subsequent development to blastocysts between the oocytes derived from vitrified follicles and those derived from fresh preantral follicles; however, the developmental competence of the oocytes derived from both vitrified and fresh preantral follicles grown in vitro was lower than that of oocytes grown in vivo. One of the five recipient mice that received 20 blastocysts derived from vitrified preantral follicles gave birth to six live pups. The results of the present study demonstrate for the first time that mouse preantral follicles can be vitrified and that some of the embryos derived from vitrified preantral follicles can develop to live pups. PMID- 11914122 TI - Effect of adrenocorticotrophic hormone on the development of oestrogen-induced changes and hyperplasia formation in the mouse uterus. AB - The aim of this study was to examine the role of adrenocorticotrophic hormone (ACTH) in proliferative and morphogenetic reactions in the uterus under continuous oestrogen treatment. Ovariectomized mice received injections of oestradiol dipropionate or vehicle once a week (2 microg per 100 g), and injections of ACTH (10 microg per 100 g) once a day or once a week for 30 days. Additional control mice received oestradiol and saline, vehicle of oestradiol, and ACTH once a day or once a week, or vehicle of ACTH, for 30 days. This study shows for the first time that ACTH affects oestrogen-dependent reactions in the uterus. Treatment with ACTH once a day resulted in a decrease in uterine mass, in cell proliferation (assessed by the number of mitotic and bromodeoxyuridine (BrdU)-labelled cells) and in the incidence of endometrial hyperplasia, in particular complex and atypical hyperplasia. Treatment with ACTH once a week led to a marked reduction in the incidence of endometrial hyperplasia, a slight increase in uterine mass and had almost no effect on cell proliferation. Daily treatment with ACTH reduced the concentration of oestrogen receptor alpha in all uterine compartments, but weekly ACTH administration had the opposite effect. Expression of glucocorticoid receptors, beta-catenin and glycogen synthase kinase 3beta in uterine tissues was lower in animals treated with oestradiol and ACTH once a day or once a week. When olive oil was used instead of oestradiol, treatments with ACTH did not produce detectable changes in all parameters examined. Thus, glucocorticoid receptor, oestrogen receptor alpha, beta-catenin and glycogen synthase kinase-3beta are involved in the effects of ACTH on oestrogen-induced changes in uterine mass, cell proliferation and the incidence of hyperplasia. PMID- 11914123 TI - Ca2+-mediated activation of ERK in hepatocytes by norepinephrine and prostaglandin F2 alpha: role of calmodulin and Src kinases. AB - BACKGROUND: Previous studies have shown that several agents that stimulate heptahelical G-protein coupled receptors activate the extracellular signal regulated kinases ERK1 (p44mapk) and ERK2 (p42mapk) in hepatocytes. The molecular pathways that convey their signals to ERK1/2 are only partially clarified. In the present study we have explored the role of Ca2+ and Ca2+-dependent steps leading to ERK1/2 activation induced by norepinephrine and prostaglandin (PG)F2alpha. RESULTS: Pretreatment of the cells with the Ca2+ chelators BAPTA-AM or EGTA, as well as the Ca2+ influx inhibitor gadolinium, resulted in a partial decrease of the ERK response. Furthermore, the calmodulin antagonists W-7, trifluoperazine, and J-8 markedly decreased ERK activation. Pretreatment with KN-93, an inhibitor of the multifunctional Ca2+/calmodulin-dependent protein kinase, had no effect on ERK activation. The Src kinase inhibitors PP1 and PP2 partially diminished the ERK responses elicited by both norepinephrine and PGF2alpha. CONCLUSION: The present data indicate that Ca2+ is involved in ERK activation induced by hormones acting on G protein-coupled receptors in hepatocytes, and suggest that calmodulin and Src kinases might play a role in these signaling pathways. PMID- 11914125 TI - The new anti-actin agent dihydrohalichondramide reveals fenestrae-forming centers in hepatic endothelial cells. AB - BACKGROUND: Liver sinusoidal endothelial cells (LSECs) react to different anti actin agents by increasing their number of fenestrae. A new structure related to fenestrae formation could be observed when LSECs were treated with misakinolide. In this study, we investigated the effects of two new actin-binding agents on fenestrae dynamics. High-resolution microscopy, including immunocytochemistry and a combination of fluorescence- and scanning electron microscopy was applied. RESULTS: Halichondramide and dihydrohalichondramide disrupt microfilaments within 10 minutes and double the number of fenestrae in 30 minutes. Dihydrohalichondramide induces fenestrae-forming centers, whereas halichondramide only revealed fenestrae-forming centers without attached rows of fenestrae with increasing diameter. Correlative microscopy showed the absence of actin filaments (F-actin) in sieve plates and fenestrae-forming centers. Comparable experiments on umbilical vein endothelial cells and bone marrow sinusoidal endothelial cells revealed cell contraction without the appearance of fenestrae or fenestrae forming centers. CONCLUSION: (I) A comparison of all anti-actin agents tested so far, revealed that the only activity that misakinolide and dihydrohalichondramide have in common is their barbed end capping activity; (II) this activity seems to slow down the process of fenestrae formation to such extent that it becomes possible to resolve fenestrae-forming centers; (III) fenestrae formation resulting from microfilament disruption is probably unique to LSECs. PMID- 11914124 TI - CCAAT/enhancer binding protein alpha uses distinct domains to prolong pituitary cells in the growth 1 and DNA synthesis phases of the cell cycle. AB - BACKGROUND: A number of transcription factors coordinate differentiation by simultaneously regulating gene expression and cell proliferation. CCAAT/enhancer binding protein alpha (C/EBPalpha) is a basic/leucine zipper transcription factor that integrates transcription with proliferation to regulate the differentiation of tissues involved in energy balance. In the pituitary, C/EBPalpha regulates the transcription of a key metabolic regulator, growth hormone. RESULTS: We examined the consequences of C/EBPalpha expression on proliferation of the transformed, mouse GHFT1-5 pituitary progenitor cell line. In contrast to mature pituitary cells, GHFT1-5 cells do not contain C/EBPalpha. Ectopic expression of C/EBPalpha in the progenitor cells resulted in prolongation of both growth 1 (G1) and the DNA synthesis (S) phases of the cell cycle. Transcription activation domain 1 and 2 of C/EBPalpha were required for prolongation of G1, but not of S. Some transcriptionally inactive derivatives of C/EBPalpha remained competent for G1 and S phase prolongation. C/EBPalpha deleted of its leucine zipper dimerization functions was as effective as full-length C/EBPalpha in prolonging G1 and S. CONCLUSION: We found that C/EBPalpha utilizes mechanistically distinct activities to prolong the cell cycle in G1 and S in pituitary progenitor cells. G1 and S phase prolongation did not require that C/EBPalpha remained transcriptionally active or retained the ability to dimerize via the leucine zipper. G1, but not S, arrest required a domain overlapping with C/EBPalpha transcription activation functions 1 and 2. Separation of mechanisms governing proliferation and transcription permits C/EBPalpha to regulate gene expression independently of its effects on proliferation. PMID- 11914126 TI - Hedgehog signal transduction proteins: contacts of the Fused kinase and Ci transcription factor with the kinesin-related protein Costal2. AB - BACKGROUND: Hedgehog signaling proteins play important roles in development by controlling growth and patterning in various animals including Drosophila and mammals. Hedgehog signaling triggers changes in responsive cells through a novel transduction mechanism that ultimately controls the transcription of specific target genes via the activity of zinc finger transcription factors of the Cubitus interruptus/GLI family. In flies, key Hedgehog signal transduction components have been identified including the kinesin-related protein Costal2, the serinethreonine kinase Fused, and the PEST-containing protein Suppressor of Fused. These proteins control Cubitus interruptus cleavage, nucleo-cytoplasmic localization and activation. In fly embryos, Costal2, Fused, Suppressor of Fused and Cubitus interruptus are associated in at least one cytoplasmic complex, which interacts with the microtubules in a Hedgehog-dependent manner. RESULTS: Here we identified and mapped direct interactions between Cos2, Fu, and Ci using an in vitro affinity assay and the yeast two-hybrid system. CONCLUSIONS: Our results provide new insights into the possible mechanism of the cytosolic steps of Hedgehog transduction. PMID- 11914127 TI - Sequence permutations in the molecular evolution of DNA methyltransferases. AB - BACKGROUND: DNA methyltransferases (MTases), unlike MTases acting on other substrates, exhibit sequence permutation. Based on the sequential order of the cofactor-binding subdomain, the catalytic subdomain, and the target recognition domain (TRD), several classes of permutants have been proposed. The majority of known DNA MTases fall into the alpha, beta, and gamma classes. There is only one member of the zeta class known and no members of the delta and epsilon classes have been identified to date. Two mechanisms of permutation have been proposed: one involving gene duplication and in-frame fusion, and the other involving inter and intragenic shuffling of gene segments. RESULTS: Two novel cases of sequence permutation in DNA MTases implicated in restriction-modification systems have been identified, which suggest that members of the delta and zeta classes (M.MwoI and M.TvoORF1413P, respectively) evolved from beta-class MTases. This is the first identification of the delta-class MTase and the second known zeta-class MTase (the first zeta-class member among DNA:m4C and m6A-MTases). CONCLUSIONS: Fragmentation of a DNA MTase gene may result from attack of nucleases, for instance when the RM system invades a new cell. Its reassembly into a functional form, the order of motifs notwithstanding, may be strongly selected for, if the cognate ENase gene remains active and poses a threat to the host's chromosome. The "cut-and-paste" mechanism is proposed for beta-delta permutation, which is non-circular and involves relocation of one segment of a gene. The circular beta zeta permutation may be explained both by gene duplication or shuffling of gene fragments. These two mechanisms are not mutually exclusive and probably both played a role in the evolution of permuted DNA MTases. PMID- 11914128 TI - Phylogenetic relationships between Hapalemur species and subspecies based on mitochondrial DNA sequences. AB - BACKGROUND: Phylogenetic relationships of the genus Hapalemur remains controversial, particularly within the Hapalemur griseus species group. In order to obtain more information on the taxonomic status within this genus, and particularly in the cytogenetic distinct subspecies group of Hapalemur griseus, 357 bp sequence of cytochrome b and 438 bp of 12S mitochondrial DNAs were analyzed on a sample of animals captured in areas extending from the north to the south-east of Madagascar. This sample covers all cytogenetically defined types recognized of the genus Hapalemur. RESULTS: Phylogenetic trees and distances analyses demonstrate a first emergence of Hapalemur simus followed by H. aureus which is the sister clade of the H. griseus subspecies. Hapalemur griseus is composed of 4 subspecies separated into two clades. The first contains H. g. griseus, H. g. alaotrensis and H. g. occidentalis. The second consists of H. g. meridionalis. A new chromosomal polymorphic variant from the region of Ranomafana, H. griseus ssp, has been analysed and was found in both clades. CONCLUSIONS: Our results support the raising of H. g. meridionalis to the specific rank H. meridionalis, while neither cytogenetic nor molecular evidences support the raising of H. g. alaotrensis to a species rank despite its morphological characteristics. The new cytotype H. g. ssp which has been previously characterized by cytogenetic studies contains animals clustering either with the group of Hapalemur griseus griseus or with that of Hapalemur meridionalis. This suggests the existence of an ancestral polymorphism or an introgression of mitochondrial DNA between subspecies. PMID- 11914129 TI - Evidence for the adaptive significance of an LTR retrotransposon sequence in a Drosophila heterochromatic gene. AB - BACKGROUND: The potential adaptive significance of transposable elements (TEs) to the host genomes in which they reside is a topic that has been hotly debated by molecular evolutionists for more than two decades. Recent genomic analyses have demonstrated that TE fragments are associated with functional genes in plants and animals. These findings suggest that TEs may contribute significantly to gene evolution. RESULTS: We have analyzed two transposable elements associated with genes in the sequenced Drosophila melanogaster y; cn bw sp strain. A fragment of the Antonia long terminal repeat (LTR) retrotransposon is present in the intron of Chitinase 3 (Cht3), a gene located within the constitutive heterochromatin of chromosome 2L. Within the euchromatin of chromosome 2R a full-length Burdock LTR retrotransposon is located immediately 3' to cathD, a gene encoding cathepsin D. We tested for the presence of these two TE/gene associations in strains representing 12 geographically diverse populations of D. melanogaster. While the cathD insertion variant was detected only in the sequenced y; cn bw sp strain, the insertion variant present in the heterochromatic Cht3 gene was found to be fixed throughout twelve D. melanogaster populations and in a D. mauritiana strain suggesting that it maybe of adaptive significance. To further test this hypothesis, we sequenced a 685bp region spanning the LTR fragment in the intron of Cht3 in strains representative of the two sibling species D. melanogaster and D. mauritiana (approximately 2.7 million years divergent). The level of sequence divergence between the two species within this region was significantly lower than expected from the neutral substitution rate and lower than the divergence observed between a randomly selected intron of the Drosophila Alcohol dehydrogenase gene (Adh). CONCLUSIONS: Our results suggest that a 359 bp fragment of an Antonia retrotransposon (complete LTR is 659 bp) located within the intron of the Drosophila melanogaster Cht3 gene is of adaptive evolutionary significance. Our results are consistent with previous suggestions that the presence of TEs in constitutive heterochromatin may be of significance to the expression of heterochromatic genes. PMID- 11914130 TI - Multiple telophase arrest bypassed (tab) mutants alleviate the essential requirement for Cdc15 in exit from mitosis in S. cerevisiae. AB - BACKGROUND: The Mitotic Exit Network (MEN) proteins - including the protein kinase Cdc15 and the protein phosphatase Cdc14 - are essential for exit from mitosis in Saccharomyces cerevisiae. To identify downstream targets of the MEN, we sought telophase arrest bypassed (tab) mutations that bypassed the essential requirement for CDC15. Previous studies identified net1(tab2-1) and CDC14(TAB6-1) as mutations in the RENT complex subunits Net1 and Cdc14, respectively, and revealed that the MEN acts by promoting release of Cdc14 from its nucleolar Net1 anchor during anaphase. However, the remaining tab mutants were not characterized. RESULTS: Fourteen out of fifteen tab mutants were mapped to three recessive (tab1-tab3) and three dominant (TAB5-TAB7) linkage groups. We show that net1(tab2-1) enables growth of tem1Delta, cdc15Delta, dbf2Delta dbf20Delta, and mob1Delta, but not cdc5Delta or cdc14Delta, arguing that whereas the essential task of the first four genes is to promote exit from mitosis, CDC5 possesses additional essential function(s). net1(tab2-1) but not CDC14(TAB6-1) resulted in a high rate of chromosome loss, indicating that Net1 promotes accurate chromosome segregation in addition to its multiple known roles. Finally, TAB1 was shown to be MTR10, a gene encoding nuclear transport receptor/adaptor. In some of the tab mutants including mtr10(tab1-1), defective nuclear export of the ribosomal protein Rpl11b was observed. Furthermore, the transport-defective -31 allele of the karyopherin SRP1, but not the transport competent -49 allele, exhibited a tab phenotype. CONCLUSIONS: Transport-defective mutations in two karyopherins can bypass cdc15Delta, suggesting that the function of the MEN is to promote mitotic exit by regulating nuclear transport. PMID- 11914131 TI - Classification and evolutionary history of the single-strand annealing proteins, RecT, Redbeta, ERF and RAD52. AB - BACKGROUND: The DNA single-strand annealing proteins (SSAPs), such as RecT, Redbeta, ERF and Rad52, function in RecA-dependent and RecA-independent DNA recombination pathways. Recently, they have been shown to form similar helical quaternary superstructures. However, despite the functional similarities between these diverse SSAPs, their actual evolutionary affinities are poorly understood. RESULTS: Using sensitive computational sequence analysis, we show that the RecT and Redbeta proteins, along with several other bacterial proteins, form a distinct superfamily. The ERF and Rad52 families show no direct evolutionary relationship to these proteins and define novel superfamilies of their own. We identify several previously unknown members of each of these superfamilies and also report, for the first time, bacterial and viral homologs of Rad52. Additionally, we predict the presence of aberrant HhH modules in RAD52 that are likely to be involved in DNA-binding. Using the contextual information obtained from the analysis of gene neighborhoods, we provide evidence of the interaction of the bacterial members of each of these SSAP superfamilies with a similar set of DNA repair/recombination protein. These include different nucleases or Holliday junction resolvases, the ABC ATPase SbcC and the single-strand-binding protein. We also present evidence of independent assembly of some of the predicted operons encoding SSAPs and in situ displacement of functionally similar genes. CONCLUSIONS: There are three evolutionarily distinct superfamilies of SSAPs, namely the RecT/Redbeta, ERF, and RAD52, that have different sequence conservation patterns and predicted folds. All these SSAPs appear to be primarily of bacteriophage origin and have been acquired by numerous phylogenetically distant cellular genomes. They generally occur in predicted operons encoding one or more of a set of conserved DNA recombination proteins that appear to be the principal functional partners of the SSAPs. PMID- 11914133 TI - Production of diamino propionic acid ammonia lyase by a new strain of Salmonella typhimurium PU011. AB - BACKGROUND: Seeds of the legume plant Lathyrus sativus, which is grown in arid and semi arid tropical regions, contain Diamino Propionic acid (DAP). DAP is a neurotoxin, which, when consumed, causes a disease called Lathyrism. Lathryrism may manifest as Neurolathyrism or Osteolathyrism, in which the nervous system, and bone formation respectively, are affected. DAP ammonia lyase is produced by a few microorganisms such as Salmonella typhi, Salmonella typhimurium and Pseudomonas, and is capable of detoxifying DAP. RESULTS: S. typhimurium PU011, a non-virulent bacterial strain isolated in our lab, was found to produce DAP ammonia lyase enzyme when grown in minimal medium containing DAP. There was a direct correlation between biomass yield and enzyme activity, until 16 h post inoculation in minimal medium containing DAP. Following ammonium sulphate precipitation and passing through Sephadex G100, CM-Sephadex and DEAE-Sephacel for crude enzyme extract preparation, about 68-fold enzyme purity was obtained. The purified enzyme gave maximum activity at pH 8.0 and was stable up to 45 degrees C. The Km value for the substrate was found to be 0.685 mM, calculated from a Line Weaver Burk plot. CONCLUSION: A new bacterial strain, S.typhimurium PU 011, which is capable of producing DAP ammonia lyase, was isolated. PMID- 11914132 TI - Auto-protective redox buffering systems in stimulated macrophages. AB - BACKGROUND: Macrophages, upon encounter with micro-organisms or stimulated by cytokines, produce various effector molecules aimed at destroying the foreign agents and protecting the organism. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are front line molecules exerting strong cytotoxic activities against micro-organisms and many cells, including macrophages themselves. Using cells of the murine macrophage cell line (RAW 264.7) stimulated in vitro with lipopolysaccharide (LPS) and/or interferon (IFN-gamma), which induce strong endogenous NO production, we examined by which mechanisms a fraction of activated macrophages protect themselves from nitrosative stress and manage to escape destruction? RESULTS: We observed that survivors (10-50% depending on the experiments) had acquired a resistant phenotype being capable to survive when further exposed in vitro to an apoptosis inducing dose of the NO donor compound DETA-NO. These cells expressed an increased steady-state levels of Mn SOD, CuZn SOD and catalase mRNA (130-200%), together with an increased activity of the corresponding enzymes. Intracellular concentration of glutathione was also increased (x 3.5 fold at 6 hours, still maintained x 5.2 fold at 48 hours). Neither mRNA for glutathione peroxydase, gamma-glutamylcysteine synthase and glutathione reductase, nor thioredoxine and thioredoxine reductase, were significantly modified. Additional experiments in which RAW 264.7 cells were stimulated with LPS and/or IFN-gamma in the presence of relatively specific inhibitors of both Mn and Cu/Zn SOD, aminotriazol (ATZ) catalase inhibitor and buthionine sulfoximine (BSO) glutathione inhibitor, showed that inhibiting LPS induced up-regulation of intracellular redox buffering systems also prevented acquisition of the resistant phenotype. CONCLUSIONS: Our data suggest a direct causal relationship between survival of a fraction of macrophages and a up regulation of key sets of auto-protective intracellular redox buffering systems, occurring simultaneously with modulation of expression of apoptotic molecules of the Bcl2-Bcl-XL/Bax-Bad family. PMID- 11914134 TI - Subunit modification and association in VR1 ion channels. AB - BACKGROUND: The capsaicin (vanilloid) receptor, VR1, is an agonist-activated ion channel expressed by sensory neurons that serves as a detector of chemical and thermal noxious stimuli. RESULTS: In the present study we investigated the properties of VR1 ion channels expressed in Xenopus oocytes. A VR1 subunit with a FLAG epitope tag at the C-terminus was constructed. When examined for size on an SDS gel, VR1-expressing oocytes produced a doublet corresponding to the size of the monomer and a band at about twice the molecular weight of the monomer. A consensus site for N-linked glycosylation was identified in the primary sequence at position 604. In channels in which the putative glycosylation site was mutated from asparagine to serine (N604S), the larger of the two monomer bands could no longer be detected on the gel. Electrophysiological experiments showed these unglycosylated channels to be functional. The high molecular weight band observed on the gel could represent either a dimer or a monomer conjugated to an unknown factor. To distinguish between these possibilities, we coexpressed a truncated VR1 subunit with full-length VR1. A band of intermediate molecular weight (composed of one full-length and one truncated subunit) was observed. This dimer persisted under strongly reducing conditions, was not affected by capsaicin or calcium, and was refractory to treatment with transglutaminase inhibitors. CONCLUSIONS: The persistence of this dimer even under harsh denaturing and reducing conditions indicates a strong interaction among pairs of subunits. This biochemical dimerization is particularly intriguing given that functional channels are almost certainly tetramers. PMID- 11914135 TI - Antinociceptive and anti-inflammatory effects of Crocus sativus L. stigma and petal extracts in mice. AB - BACKGROUND: Crocus sativus L. (saffron) is used in folk medicine, for example as an antiedematogenic agent. We aimed to evaluate the antinociceptive and anti inflammatory activity of saffron extracts in mice. RESULTS: We used aqueous and ethanolic maceration extracts of Crocus sativus L. stigma and petals. Antinociceptive activity was examined using the hot plate and writhing tests. The effect of extracts against acute inflammation was studied using xylene induced ear edema in mice. The activity of the extracts against chronic inflammation was assessed by formalin-induced edema in the rat paw. In the hot plate tests, intraperitoneal injection of both extracts showed no significant antinociceptive activity in mice. The extracts exhibited antinociceptive activity against acetic acid induced writhing. Naloxone partially blocked only the antinociceptive activity of the stigma aqueous extract. Only the stigma extracts showed weak to moderate effect against acute inflammation. In chronic inflammation, both aqueous and ethanolic stigma extracts, as well as ethanolic petal extract, exerted anti inflammatory effects. CONCLUSIONS: We conclude that aqueous and ethanolic extracts of saffron stigma and petal have an antinociceptive effect, as well as acute and/or chronic anti-inflammatory activity. PMID- 11914136 TI - T Wave Alternans in high arrhythmic risk patients: analysis in time and frequency domains: a pilot study. AB - BACKGROUND: T wave alternans (TA) is a repolarisation phenomenon manifesting as a microvolt beat to beat change in the amplitude of the T wave and ST segment. TA has been shown to be a predictor of arrhythmic risk in unselected myocardial infarction populations. TA has not been used to differentiate risk within the ischaemic cardiomyopathy population. METHODS: The subjects investigated comprised, Group 1: 7 stable patients with remote (>20 months) extensive myocardial scarring and no arrhythmic events (NYHA 3 and 4). Group2: 9 post infarction patients with malignant arrhythmia and implantable defibrillator. During breath holding, 20 continuous QRST complexes from each patients X, Y and Z leads were digitally recorded. Time domain, resultant absolute difference vectors (ATA), were calculated for alternate resultant T wave sequences. Group differences between the magnitude and temporal distribution of mean ATAs and their spectral and cross-spectral analysis were compared. RESULTS: Group 1 v Group 2 mean ATAs were 10.7 (7.17) v 11.7 (8.48) respectively, not significant. Each group had a homogenous temporal distribution of ATAs. Both group's largest mean ATA frequency components were between 0 to 25 Hz, the largest ATA component being at the DC frequency. Cross spectral analysis showed no significant differences in group ATA frequency content. CONCLUSION: The frequency content and microvolt magnitude of T wave alternans was not significantly different in these two groups. The specificity of T wave alternans for differentiating arrhythmic risk in post infarction scarring and heart failure needs investigation. PMID- 11914137 TI - Markers for early detection of cancer: statistical guidelines for nested case control studies. AB - BACKGROUND: Recently many long-term prospective studies have involved serial collection and storage of blood or tissue specimens. This has spurred nested case control studies that involve testing some specimens for various markers that might predict cancer. Until now there has been little guidance in statistical design and analysis of these studies. METHODS: To develop statistical guidelines, we considered the purpose, the types of biases, and the opportunities for extracting additional information. RESULTS: The following guidelines: (1) For the clearest interpretation, statistics should be based on false and true positive rates - not odds ratios or relative risks (2) To avoid overdiagnosis bias, cases should be diagnosed as a result of symptoms rather than on screening. (3) To minimize selection bias, the spectrum of control conditions should be the same in study and target screening populations. (4) To extract additional information, criteria for a positive test should be based on combinations of individual markers and changes in marker levels over time. (5) To avoid overfitting, the criteria for a positive marker combination developed in a training sample should be evaluated in a random test sample from the same study and, if possible, a validation sample from another study. (6) To identify biomarkers with true and false positive rates similar to mammography, the training, test, and validation samples should each include at least 110 randomly selected subjects without cancer and 70 subjects with cancer. CONCLUSION: These guidelines ensure good practice in the design and analysis of nested case-control studies of early detection biomarkers. PMID- 11914138 TI - The probability of cost-effectiveness. AB - BACKGROUND: The study of cost-effectiveness comparisons between competing medical interventions has led to a variety of proposals for quantifying cost effectiveness. The differences between the various approaches can be subtle, and one purpose of this article is to clarify some important distinctions. DISCUSSION: We discuss alternative measures in the framework of individual, patient-level, incremental net benefits. In particular we examine the probability of cost-effectiveness for an individual, proposed by Willan. SUMMARY: We argue that this is a useful addition to the range of cost-effectiveness measures, but will be of secondary interest to most decision makers. We also demonstrate that Willan's proposed estimate of this probability is logically flawed. PMID- 11914141 TI - Endovascular treatment of huge saccular abdominal aortic aneurysm in a young Behcet patient: mid-term result. AB - BACKGROUND: Abdominal aortic aneurysm formation is among the arterial complications of Behcet's disease. Weakness and fragility of aortic walls leads to the development of arterial complications like pseudoaneurysms. CASE PRESENTATION: A case of huge saccular abdominal aortic aneurysm in a young Behcet patient who was successfully treated with endovascular stent graft placement is reported, diagnostic and interventional procedures are discussed, and mid-term follow-up results are presented. CONCLUSIONS: Endovascular treatment of abdominal aortic aneurysm complications of young Behcet patients who are not suitable for open surgery and need intervention could be an alternative treatment modality even without performing preprocedural angiography. PMID- 11914140 TI - Gastric adenocarcinoma in a patient re-infected with H. pylori after regression of MALT lymphoma with successful anti-H. pylori therapy and gastric resection: a case report. AB - BACKGROUND: Helicobacter pylori (H. pylori) has been etiologically linked with primary gastric lymphoma (PGL) and gastric carcinoma (GC). There are a few reports of occurrence of both diseases in the same patients with H. pylori infection. CASE PRESENTATION: We report a patient with PGL in whom the tumor regressed after surgical resection combined with eradication of H. pylori infection. However, he developed GC on follow up; this was temporally associated with recrudescence/re-infection of H. pylori. This is perhaps first report of such occurrence. CONCLUSIONS: Possible cause and effect relationship between H. pylori infection and both PGL and GC is discussed. This case also documents a unique problem in management of PGL in tropical countries where re-infection with H. pylori is supposed to be high. PMID- 11914139 TI - Primary biliary cirrhosis and autoimmune cholangitis are not associated with coeliac disease in Crete. AB - BACKGROUND: An increased prevalence of coeliac disease in patients with primary biliary cirrhosis has been recently reported. However, in other studies the association has not been confirmed. There have been no formal attempts to systematically evaluate patients with autoimmune cholangitis for coeliac disease. METHODS: Sera from 62 patients with primary biliary cirrhosis, 17 with autoimmune cholangitis and 100 blood donors were screened for anti-gliadin, anti-endomysial, anti-reticulin, and IgA class antibodies to guinea pig liver-derived tissue transglutaminase. Eighteen untreated coeliacs served as methodological controls. Analyses were performed by using the chi2 and Fischer's exact tests. RESULTS: Anti-gliadin antibodies were detected in 21% of patients with primary biliary cirrhosis, 35% of patients with autoimmune cholangitis, and 3% of controls (p < 0.001). IgA class gliadin antibodies positivity was more pronounced in patients with Scheuer's stage III-IV disease (p < 0.05). Anti-transglutaminase antibodies were detected in 10% and in 18% of patients with primary biliary cirrhosis and autoimmune cholangitis respectively (p < 0.001). Anti-reticulin and anti endomysial antibodies were negative in all patients. Duodenal biopsies were performed in 59% and 71% of patients with primary biliary cirrhosis and autoimmune cholangitis respectively, tested positive for at least one antibody class. No histological features of coeliac disease were found. CONCLUSIONS: We were unable to demonstrate an increased risk of coeliac disease in patients with primary biliary cirrhosis and autoimmune cholangitis. Our results confirm the previously reported high prevalence of false-positive anti-gliadin and guinea pig liver-derived anti-tissue transglutaminase antibodies in patients with chronic liver disease. PMID- 11914142 TI - Elevated serum immunoglobulin E to Cryptomeria japonica pollen in rats exposed to diesel exhaust during fetal and neonatal periods. AB - BACKGROUND: The possible participation of diesel exhaust inhalation during the fetal period in the elevation of IgE against pollen in postnatal life was investigated. METHOD: The experiment was conducted using rat pups. Group I; exposed to clean air (Control). Group 2, 3; exposed to total or filtered diesel exhaust during the fetal period (Total-C-C, Filtered-C-C). Group 4, 5; exposed to total or filtered diesel exhaust during the suckling period (C-Total-C, C Filtered-C). Group 6,7; exposed to total or filtered diesel exhaust during the weaning period (C-C-Total, C-C-Filtered). Total diesel engine exhaust contained 1.73 mg/m3 particulate matter and 0.79 ppm nitrogen dioxide; filtered exhaust contained the same gases as the total exhaust without particulate matter. Intraperitoneal injection of 5 mg crude cedar pollen was performed at 2-week intervals from the 49th day after birth. RESULTS: The mean IgE titers measured by the P-K reaction in the Control, Total-C-C, Filtered-C-C, C-Total-C, C-Filtered C, C-C-Total and C-C-Filtered were 64.0 ± 2.7, 469.5 ± 1.6, 332.0 ± 1.7, 380.4 ± 1.7, 394.8 ± 1.7, 115.9 ± 1.3 and 57.0 ± 2.8 respectively after the fourth immunization. There were significant differences between Total-C-C, Filtered-C-C, C-Total-C, C-Filtered-C and Control (p < 0.01, p < 0.01, p < 0.05, p < 0.01, respectively). The mean IgE titers by ELISA were 0.47 ± 0.06, 0.79 ± 0.35, 0.86 ± 0.46, 0.80 ± 0.22, 0.56 ± 0.08, 0.46 ± 0.04 and 0.45 ± 0.03, respectively. IgE titers in Filtered-C-C and C-Total-C were significantly higher than in Control (p < 0.05 for each). CONCLUSIONS: Inhalation of diesel exhaust during differentiation of the immune system accelerated the elevation of IgE against pollen. PMID- 11914143 TI - Chromosomal aberrations in benign and malignant bilharzia-associated bladder lesions analyzed by comparative genomic hybridization. AB - BACKGROUND: Bilharzia-associated bladder cancer (BAC) is a major health problem in countries where urinary schistosomiasis is endemic. Characterization of the genetic alterations in this cancer might enhance our understanding of the pathogenic mechanisms of the disease but, in contrast to nonbilharzia bladder cancer, BAC has rarely been the object of such scrutiny. In the present study, we aimed to characterize chromosomal imbalances in benign and malignant post bilharzial lesions, and to determine whether their unique etiology yields a distinct cytogenetic profile as compared to chemically induced bladder tumors. METHODS: DNAs from 20 archival paraffin-embedded post-bilharzial bladder lesions (6 benign and 14 malignant) obtained from Sudanese patients (12 males and 8 females) with a history of urinary bilharziasis were investigated for chromosomal imbalances using comparative genomic hybridization (CGH). Subsequent FISH analysis with pericentromeric probes was performed on paraffin sections of the same cases to confirm the CGH results. RESULTS: Seven of the 20 lesions (6 carcinomas and one granuloma) showed chromosomal imbalances varying from 1 to 6 changes. The most common chromosomal imbalances detected were losses of 1p21-31, 8p21-pter, and 9p and gain of 19p material, seen in three cases each, including the benign lesion. CONCLUSION: Most of the detected imbalances have been repeatedly reported in non-bilharzial bladder carcinomas, suggesting that the cytogenetic profiles of chemical- and bilharzia-induced carcinomas are largely similar. However, loss of 9p seems to be more ubiquitous in BAC than in bladder cancer in industrialized countries. PMID- 11914144 TI - Delta-aminolevulinic acid cytotoxic effects on human hepatocarcinoma cell lines. AB - BACKGROUND: Acute Intermittent Porphyria is a genetic disorder of heme metabolism, characterized by increased levels of porphyrin precursors, delta aminolevulinic acid (ALA) and porphobilinogen (PBG). ALA has been reported to generate reactive oxygen species and to cause oxidative damage to proteins, subcellular structures and DNA. It is known that oxidative stress can induce apoptosis. The aim of this work was to study the cytotoxic effect of ALA on two hepatocarcinoma cell lines. RESULTS: We have determined the impact of ALA on HEP G2 and HEP 3B hepatocarcinoma cell lines survival as measured by the MTT assay. ALA proved to be cytotoxic in both cell lines however; HEP G2 was more sensitive to ALA than HEP 3B. Addition of hemin or glucose diminished ALA cytotoxicity in HEP G2 cells; instead it was enhanced in HEP 3B cells. Because apoptosis is usually associated with DNA fragmentation, the DNA of ALA treated and untreated cells were analyzed. The characteristic pattern of DNA fragmentation ladders was observed in ALA treated cells. To elucidate the mechanisms of ALA induced apoptosis, we examined its effect on p53 expression. No changes in p53 mRNA levels were observed after exposure of both cell lines to ALA for 24 h. CDK2 and CDK4 protein levels were reduced after ALA treatment at physiological concentrations. PMID- 11914145 TI - Assessing the quality of reports of randomized trials in pediatric complementary and alternative medicine. AB - OBJECTIVE: To evaluate the quality of reports of complementary and alternative medicine (CAM) randomized controlled trials (RCTs) in the pediatric population. We also examined whether there was a change in the quality of reporting over time. METHODS: We used a systematic sample of 251 reports of RCTs that used a CAM intervention. The quality of each report was assessed using the number of CONSORT checklist items included, the frequency of unclear allocation concealment, and a 5-point quality assessment instrument. RESULTS: Nearly half (40%) of the CONSORT checklist items were included in the reports, with an increase in the number of items included. The majority (81.3%) of RCTs reported unclear allocation concealment with no significant change over time. The quality of reports achieved approximately 40% of their maximum possible total score as assessed with the Jadad scale with no change over time. Information regarding adverse events was reported in less than one quarter of the RCTs (22%) and information regarding costs was mentioned in only a minority of reports (4%). CONCLUSIONS: RCTs are an important tool for evidence based health care decisions. If these studies are to be relevant in the evaluation of CAM interventions it is important that they are conducted and reported with the highest possible standards. There is a need to redouble efforts to ensure that children and their families are participating in RCTs that are conducted and reported with minimal bias. Such studies will increase their usefulness to a board spectrum of interested stakeholders. PMID- 11914146 TI - Assessing the quality of reports of systematic reviews in pediatric complementary and alternative medicine. AB - OBJECTIVE: To examine the quality of reports of complementary and alternative medicine (CAM) systematic reviews in the pediatric population. We also examined whether there were differences in the quality of reports of a subset of CAM reviews compared to reviews using conventional interventions. METHODS: We assessed the quality of reports of 47 CAM systematic reviews and 19 reviews evaluating a conventional intervention. The quality of each report was assessed using a validated 10-point scale. RESULTS: Authors were particularly good at reporting: eligibility criteria for including primary studies, combining the primary studies for quantitative analysis appropriately, and basing their conclusions on the data included in the review. Reviewers were weak in reporting: how they avoided bias in the selection of primary studies, and how they evaluated the validity of the primary studies. Overall the reports achieved 43% (median = 3) of their maximum possible total score. The overall quality of reporting was similar for CAM reviews and conventional therapy ones. CONCLUSIONS: Evidence based health care continues to make important contributions to the well being of children. To ensure the pediatric community can maximize the potential use of these interventions, it is important to ensure that systematic reviews are conducted and reported at the highest possible quality. Such reviews will be of benefit to a broad spectrum of interested stakeholders. PMID- 11914147 TI - Assessment of the infectious diseases surveillance system of the Republic of Armenia: an example of surveillance in the Republics of the former Soviet Union. AB - BACKGROUND: Before 1991, the infectious diseases surveillance systems (IDSS) of the former Soviet Union (FSU) were centrally planned in Moscow. The dissolution of the FSU resulted in economic stresses on public health infrastructure. At the request of seven FSU Ministries of Health, we performed assessments of the IDSS designed to guide reform. The assessment of the Armenian infectious diseases surveillance system (AIDSS) is presented here as a prototype. DISCUSSION: We performed qualitative assessments using the Centers for Disease Control and Prevention (CDC) guidelines for evaluating surveillance systems. Until 1996, the AIDSS collected aggregate and case-based data on 64 infectious diseases. It collected information on diseases of low pathogenicity (e.g., pediculosis) and those with no public health intervention (e.g., infectious mononucleosis). The specificity was poor because of the lack of case definitions. Most cases were investigated using a lengthy, non-disease-specific case-report form Armenian public health officials analyzed data descriptively and reported data upward from the local to national level, with little feedback. Information was not shared across vertical programs. Reform should focus on enhancing usefulness, efficiency, and effectiveness by reducing the quantity of data collected and revising reporting procedures and information types; improving the quality, analyses, and use of data at different levels; reducing system operations costs; and improving communications to reporting sources. These recommendations are generalizable to other FSU republics. SUMMARY: The AIDSS was complex and sensitive, yet costly and inefficient. The flexibility, representativeness, and timeliness were good because of a comprehensive health-care system and compulsory reporting. Some data were questionable and some had no utility. PMID- 11914148 TI - Predictors of refusal to participate: a longitudinal health survey of the elderly in Australia. AB - BACKGROUND: The loss of participants in longitudinal studies due to non-contact, refusal or death can introduce bias into the results of such studies. The study described here examines reasons for refusal over three waves of a survey of persons aged >or=70 years. METHODS: In a longitudinal study involving three waves, participants were compared to those who refused to participate but allowed an informant to be interviewed and to those who refused any participation. RESULTS: At Wave 1 both groups of Wave 2 non-participants had reported lower occupational status and fewer years of education, had achieved lower verbal IQ scores and cognitive performance scores and experienced some distress from the interview. Those with an informant interview only were in poorer physical health than those who participated and those who refused. Depression and anxiety symptoms were not associated with non-participation. Multivariate analyses found that verbal IQ and cognitive impairment predicted refusal. Results were very similar for refusers at both Waves 2 and 3. CONCLUSIONS: Longitudinal studies of the elderly may over estimate cognitive performance because of the greater refusal rate of those with poorer performance. However, there is no evidence of bias with respect to anxiety or depression. PMID- 11914149 TI - Giardiasis in children living in post-earthquake camps from Armenia (Colombia). AB - BACKGROUND: An earthquake in the coffee growing region of Colombia on January 25, 1999 destroyed 70% of the houses in Armenia city. Transitory housing camps still remained until two years after the disaster. Parasitological studies found that, in this population, giardiasis was the most frequent parasitic infection. This study was carried out in order to determine the epidemiological risk factors associated with this high prevalence. METHODS: Fecal samples were obtained from 217 children aged between 3 and 13 years. Stool samples were studied by direct wet examination and stained with ferric hematoxilin for microscopical examination. Epidemiological data were collected by questionnaire and analyzed by using the Epi-info software (CDC, Atlanta 2001). RESULTS: Giardia cysts were observed in 60.4% of the samples presented and trophozoites in 4.6%. The following epidemiological and laboratory factors were significantly associated with Giardia infection: 1. Use of communal toilet (vs. individual toilet) OR: 3.9, CI95%: 1.2-16; 2. water provision by municipal ducts (vs. water provision by individual tanks) OR: 3.5, CI95% 1.1-14, and 3. presence of mucus in stool OR: 2.3, IC95%: 0.9-6.7. CONCLUSIONS: A high prevalence of giardiasis was found in children living in temporary houses after the 1999 earthquake in Armenia (Colombia). Giardiasis is an emerging disease in post-disaster situations and adequate prevention measures should be implemented during these circumstances. PMID- 11914151 TI - Is albumin gradient or fluid to serum albumin ratio better than the pleural fluid lactate dehydroginase in the diagnostic of separation of pleural effusion? AB - BACKGROUND: To determine the accuracy of serum-effusion albumin gradient (SEAG) and pleural fluid to serum albumin ratio (ALBR) in the diagnostic separation of pleural effusion into transudate and exudate and to compare SEAG and ALBR with pleural fluid LDH (FLDH) the most widely used test. METHODS: Data collected from 200 consecutive patients with a known cause of pleural effusion in a United Kingdom district general hospital. RESULTS: The median and inter quartile ranges (IQR) for SEAG 93.5 (33.8 to 122.5) g/dl, ALBR 0.49 (0.42 to 0.62) and FLDH 98.5 IU/L(76.8 to 127.5) in transudates were significantly lower than the corresponding values for exudates 308.5 (171 to 692), 0.77 (0.63 to 0.85), 344 (216 to 695) all p < 0.0001. The Area Under the Curve (AUC) with 95% confidence intervals (Cl) for SEAG, ALBR and FLDH were 0.81 (0.75 to 0.87), 0.79 (0.72 to 0.86) and 0.9 (0.87 to 0.96) respectively. The positive likelihood ratios with 95%CI for FLDH, SEAG, and ALBR were: 7.3(3.5-17), 6.3(3-15) 6.2(3-14) respectively. There was a significant negative correlation between SEAG and ALBR (r= -0.89, p < 0.0001). CONCLUSION: The discriminative value for SEAG and ALBR appears to be similar in the diagnostic separation of transudates and exudates. FLDH is a superior test compared to SEAG and ALBR. PMID- 11914152 TI - Observations on morphologic changes in the aging and degenerating human disc: secondary collagen alterations. AB - BACKGROUND: In the annulus, collagen fibers that make up the lamellae have a wavy, planar crimped pattern. This crimping plays a role in disc biomechanical function by allowing collagen fibers to stretch during compression. The relationship between morphologic changes in the aging/degenerating disc and collagen crimping have not been explored. METHODS: Ultrastructural studies were performed on annulus tissue from 29 control (normal) donors (aged newborn to 79 years) and surgical specimens from 49 patients (aged 16 to 77 years). Light microscopy and specialized image analysis to visualize crimping was performed on additional control and surgical specimens. Human intervertebral disc tissue from the annulus was obtained in a prospective morphologic study of the annulus. Studies were approved by the authors' Human Subjects Institutional Review Board. RESULTS: Three types of morphologic changes were found to alter the crimping morphology of collagen: 1) encircling layers of unusual matrix disrupted the lamellar collagen architecture; 2) collagen fibers were reduced in amount, and 3) collagen was absent in regions with focal matrix loss. CONCLUSIONS: Although proteoglycan loss is well recognized as playing a role in the decreased shock absorber function of the aging/degenerating disc, collagen changes have received little attention. This study suggests that important stretch responses of collagen made possible by collagen crimping may be markedly altered by morphologic changes during aging/degeneration and may contribute to the early tissue changes involved in annular tears. PMID- 11914150 TI - Adverse effects of the antimalaria drug, mefloquine: due to primary liver damage with secondary thyroid involvement? AB - BACKGROUND: Mefloquine is a clinically important antimalaria drug, which is often not well tolerated. We critically reviewed 516 published case reports of mefloquine adverse effects, to clarify the phenomenology of the harms associated with mefloquine, and to make recommendations for safer prescribing. PRESENTATION: We postulate that many of the adverse effects of mefloquine are a post-hepatic syndrome caused by primary liver damage. In some users we believe that symptomatic thyroid disturbance occurs, either independently or as a secondary consequence of the hepatocellular injury. The mefloquine syndrome presents in a variety of ways including headache, gastrointestinal disturbances, nervousness, fatigue, disorders of sleep, mood, memory and concentration, and occasionally frank psychosis. Previous liver or thyroid disease, and concurrent insults to the liver (such as from alcohol, dehydration, an oral contraceptive pill, recreational drugs, and other liver-damaging drugs) may be related to the development of severe or prolonged adverse reactions to mefloquine. IMPLICATIONS: We believe that people with active liver or thyroid disease should not take mefloquine, whereas those with fully resolved neuropsychiatric illness may do so safely. Mefloquine users should avoid alcohol, recreational drugs, hormonal contraception and co-medications known to cause liver damage or thyroid damage. With these caveats, we believe that mefloquine may be safely prescribed in pregnancy, and also to occupational groups who carry out safety-critical tasks. TESTING: Mefloquine's adverse effects need to be investigated through a multicentre cohort study, with small controlled studies testing specific elements of the hypothesis. PMID- 11914153 TI - Is endosonography an effective method for detection and local staging of the ampullary carcinoma? A prospective study. AB - BACKGROUND: The relatively rare carcinoma of the ampulla of Vater is a neoplasia with a good prognosis compared to pancreatic cancer. Preoperative staging is important in planning the most suitable surgical intervention. AIM: To prospectively evaluate the diagnostic accuracy of Endoscopic Ultrasonography (EUS) in comparison with conventional US and CT scan, in staging of patients with ampullary carcinoma. PATIENTS AND METHODS: 20 patients (7 women and 13 men) with histologically proven carcinoma of the ampulla of Vater were assessed by EUS, CT scan and US. Results were compared to surgical findings. RESULTS: Endoscopic biopsies were diagnostic in 76% of the patients. Detection of ampullary cancer with US and CT scan was 15% and 20% respectively. Only indirect signs of the disease were identified in the majority of cases using these methods. Overall accuracy of EUS in detection of ampullary tumours was 100%. The EUS was significantly (p < 0.001) superior than US and CT scan in ampullary carcinoma detection. Tumour size, tumour extension and the existence of metastatic lymph nodes were also identified and EUS proved to be very useful for the preoperative classification both for the T and the N components of the TNM staging of this neoplasia. The diagnostic accuracy for tumour extension (T) was 82% and for detection of metastatic lymph nodes (N) was 71%. CONCLUSION: EUS is more accurate in detecting ampullary cancer than US and CT scan. Tumor extension and locally metastatic lymph nodes are more accurately assessed by means of EUS than with other imaging methods. PMID- 11914154 TI - Treatment of disseminated granuloma annulare with fumaric acid esters. AB - BACKGROUND: Granuloma annulare is a granulomatous disease of unknown etiology. Various therapies have been tried in disseminated granuloma annulare (DGA), including corticosteroids, several variants of psoralen plus ultraviolet-A radiation, ultraviolet- A1 radiation, systemic retinoids, and dapsone, with variable success. We report a patient with recalcitrant DGA who was treated with fumaric acid esters (FAE). CASE PRESENTATION: A 40-year old Caucasian woman presented with a 25-year history of recalcitrant DGA. On both legs and the abdomen there were erythematous annular plaques. She was treated with FAE in tablet form using two formulations differing in strength (low strength tablets: 30 mg dimethylfumarate, 67 mg monoethylfumarate Ca salt, 5 mg monoethylfumarate Mg salt, 3 mg monoethylfumarate Zn salt; high strength tablets: 120 mg dimethylfumarate, 87 mg monoethylfumarate Ca salt, 5 mg monoethylfumarate Mg salt, 3 mg monoethylfumarate Zn salt). After three-month therapy, an almost complete clearance of skin lesions was achieved. With the exception of temporary lymphopenia, no adverse effects were observed. The patient remained in remission during a six-month follow up period. CONCLUSIONS: Our observation has demonstrated that FAE is a potentially beneficial therapeutic option for patients with recalcitrant DGA. However controlled trials are necessary to fully explore the efficacy, optimal dosage, and safety of FAE in the management of DGA. PMID- 11914155 TI - Rhodococcus erythropolis ATCC 25544 as a suitable source of cholesterol oxidase: cell-linked and extracellular enzyme synthesis, purification and concentration. AB - BACKGROUND: The suitability of the strain Rhodococcus erythropolis ATCC 25544 grown in a two-liter fermentor as a source of cholesterol oxidase has been investigated. The strain produces both cell-linked and extracellular cholesterol oxidase in a high amount, that can be extracted, purified and concentrated by using the detergent Triton X-114. RESULTS: A spray-dry method of preparation of the enzyme inducer cholesterol in Tween 20 was found to be superior in both convenience and enzyme synthesis yield to one of heat-mixing. Both were similar as far as biomass yield is concerned. Cell-linked cholesterol oxidase was extracted with Triton X-114, and this detergent was also used for purification and concentration, following temperature-induced detergent phase separation. Triton X-114 was utilized to purify and to concentrate the cell-linked and the extracellular enzyme. Cholesterol oxidase was found mainly in the resulting detergent-rich phase. When Triton X-114 concentration was set to 6% w/v the extracellular, but not the cell-extracted enzyme, underwent a 3.4-fold activation after the phase separation process. This result is interpreted in the light of interconvertible forms of the enzyme that do not seem to be in equilibrium. Fermentation yielded 360 U/ml (672 U/ml after activation), 36% of which was extracellular (65% after activation). The Triton X-114 phase separation step yielded 11.6-fold purification and 20.3-fold concentration. CONCLUSIONS: The results of this work may make attractive and cost-effective the implementation of this bacterial strain and this detergent in a purification-based industrial production scheme of commercial cholesterol oxidase. PMID- 11914156 TI - The role of soil factors and leaf protein in the utilization of mopane plants by elephants in northern Botswana. AB - BACKGROUND: Mopane (Colophospermum mopane) plants form monotypic woodlands that cover extensive areas in northern Botswana. Mopane is also a principal food item in the diet of elephants. Obtrusive damage to mopane plants as a result of elephant feeding may alter the structure of mopane woodlands. Some mopane woodland areas in northern Botswana are subjected to heavy elephant utilization rates whereas other mopane areas are less affected. However, the underlying reason for the concentrated elephant utilization is unknown. RESULTS: Ten mopane plots were subjected to sampling of soil properties that included structure, pH, nitrogen, phosphorus, potassium contents and protein contents. Elevated nitrogen and phosphorus contents in soils correlated with high protein levels in mopane leaves. Protein levels in leaves of mopane plants differed significantly between sites. However, multivariate analyses of environmental parameters and plots suggested that on a regional scale, there was no difference in the extent of elephant damage to mopane plants due to differential protein levels in leaves or any of the underlying soi factors that were examined. CONCLUSIONS: From management perspective, this pattern mitigates the likelihood that an even more prolific elephant population will alter mopane woodland habitats irreversibly. PMID- 11914157 TI - Salmon-derived nitrogen in terrestrial invertebrates from coniferous forests of the Pacific Northwest. AB - BACKGROUND: Bi-directional flow of nutrients between marine and terrestrial ecosystems can provide essential resources that structure communities in transitional habitats. On the Pacific coast of North America, anadromous salmon (Oncorhynchus spp.) constitute a dominant nutrient subsidy to aquatic habitats and riparian vegetation, although the contribution to terrestrial habitats is not well established. We use a dual isotope approach of delta15N and delta13C to test for the contribution of salmon nutrients to multiple trophic levels of litter based terrestrial invertebrates below and above waterfalls that act as a barrier to salmon migration on two watersheds in coastal British Columbia. RESULTS: Invertebrates varied predictably in delta15N with enrichment of 3-8 per thousand below the falls compared with above the falls in all trophic groups on both watersheds. We observed increasing delta15N levels in our invertebrate groups with increasing consumption of dietary protein. Invertebrates varied in delta13C but did not always vary predictably with trophic level or habitat. From 19.4 to 71.5% of invertebrate total nitrogen was originally derived from salmon depending on taxa, watershed, and degree of fractionation from the source. CONCLUSIONS: Enrichment of delta15N in the invertebrate community below the falls in conjunction with the absence of delta13C enrichment suggests that enrichment in delta15N occurs primarily through salmon-derived nitrogen subsidies to litter, soil and vegetation N pools rather than from direct consumption of salmon tissue or salmon tissue consumers. Salmon nutrient subsidies to terrestrial habitats may result in shifts in invertebrate community structure, with subsequent implications for higher vertebrate consumers, particularly the passerines. PMID- 11914158 TI - Glucosinolate breakdown products as insect fumigants and their effect on carbon dioxide emission of insects. AB - BACKGROUND: Glucosinolate breakdown products are volatile, therefore good candidates for insect fumigants. However, although they are insecticidal, the mode of action of such natural products is not clear. We studied the insecticidal effect of these compounds as fumigants, and monitored the production of carbon dioxide by the insects as a probe to the understanding of their mode of action. RESULTS: The fumigation 24-h LC50 against the house fly (Musca domestica L.) of allyl thiocyanate, allyl isothiocyanate, allyl cyanide, and l-cyano-2-hydroxy-3 butene was 0.1, 0.13, 3.66, and 6.2 microg cm-3, respectively; they were 0.55, 1.57, 2.8, and > 19.60 microg cm-3, respectively, against the lesser grain borer (Rhyzopertha dominica Fabricius). The fumigation toxicity of some of the glucosinolate products was very close to or better than that of the commercial insect fumigants such as chloropicrin (LC50: 0.08 and 1.3 microg cm-3 against M. domestica and R. dominica, respectively) and dichlorovos (LC50: < 0.02 and 0.29 microg cm-3 against M. domestica and R. dominica, respectively) in our laboratory tests. Significantly increased CO2 expiration was found in insects exposed to the vapor of allyl isothiocyanate, allyl thiocyanate and allyl isocyanate. Allyl isothiocyanate was also found to increase the CO2 expiration of the American cockroach (Periplaneta americana L.). CONCLUSIONS: Glucosinolate breakdown products have potential as biodegradable and safe insect fumigants. They may act on the insect respiratory system in their mode of action. PMID- 11914160 TI - Electroacupuncture versus diclofenac in symptomatic treatment of osteoarthritis of the knee: a randomized controlled trial. AB - BACKGROUND: The purpose of this study was to compare the efficacy of electroacupuncture (EA), diclofenac and their combination in symptomatic treatment of osteoarthritis (OA) of the knee. METHODS: This study was a randomized, single-blind, placebo controlled trial. The 193 out-patients with OA of the knee were randomized into four groups: placebo, diclofenac, EA and combined (diclofenac plus EA). Paracetamol tablets were prescribed as a rescue analgesic during the study. The patients were evaluated after a run-in period of one week (week 0) and again at the end of the study (week 4). The clinical assessments included the amount of paracetamol taken/week, visual analog scale (VAS), Western Ontario and McMaster Universities (WOMAC) OA Index, Lequesne's functional index, 50 feet-walk time, and the orthopedist's and patient's opinion of change. RESULTS: One hundred and eighty six patients completed the study. The improvement of symptoms (reduction in mean changes) in most outcome parameters was greatest in the EA group. The proportions of responders and patients with an overall opinion of "much better" were also greatest in the EA group. The improvement in VAS was significantly different between the EA and placebo group as well as the EA and diclofenac group. The improvement in Lequesne's functional index also differed significantly between the EA and placebo group. In addition, there was a significant improvement in WOMAC pain index between the combined and placebo group. CONCLUSION: EA is significantly more effective than placebo and diclofenac in the symptomatic treatment of OA of the knee in some circumstances. However, the combination of EA and diclofenac treatment was no more effective than EA treatment alone. PMID- 11914159 TI - Tetraethylammonium block of water flux in Aquaporin-1 channels expressed in kidney thin limbs of Henle's loop and a kidney-derived cell line. AB - BACKGROUND: Aquaporin-1 (AQP1) channels are constitutively active water channels that allow rapid transmembrane osmotic water flux, and also serve as cyclic-GMP gated ion channels. Tetraethylammonium chloride (TEA; 0.05 to 10 mM) was shown previously to inhibit the osmotic water permeability of human AQP1 channels expressed in Xenopus oocytes. The purpose of the present study was to determine if TEA blocks osmotic water flux of native AQP1 channels in kidney, and recombinant AQP1 channels expressed in a kidney derived MDCK cell line. We also demonstrate that TEA does not inhibit the cGMP-dependent ionic conductance of AQP1 expressed in oocytes, supporting the idea that water and ion fluxes involve pharmacologically distinct pathways in the AQP1 tetrameric complex. RESULTS: TEA blocked water permeability of AQP1 channels in kidney and kidney-derived cells, demonstrating this effect is not limited to the oocyte expression system. Equivalent inhibition is seen in MDCK cells with viral-mediated AQP1 expression, and in rat renal descending thin limbs of Henle's loops which abundantly express native AQP1, but not in ascending thin limbs which do not express AQP1. External TEA (10 mM) does not block the cGMP-dependent AQP1 ionic conductance, measured by two-electrode voltage clamp after pre-incubation of oocytes in 8Br-cGMP (10-50 mM) or during application of the nitric oxide donor, sodium nitroprusside (2-4 mM). CONCLUSIONS: TEA selectively inhibits osmotic water permeability through native and heterologously expressed AQP1 channels. The pathways for water and ions in AQP1 differ in pharmacological sensitivity to TEA, and are consistent with the idea of independent solute pathways within the channel structure. The results confirm the usefulness of TEA as a pharmacological tool for the analysis of AQP1 function. PMID- 11914161 TI - A randomised controlled trial of a patient based Diabetes Recall and Management System: the DREAM trial: a study protocol [ISRCTN32042030]. AB - BACKGROUND: Whilst there is broad agreement on what constitutes high quality health care for people with diabetes, there is little consensus on the most efficient way of delivering it. Structured recall systems can improve the quality of care but the systems evaluated to date have been of limited sophistication and the evaluations have been carried out in small numbers of relatively unrepresentative settings. Hartlepool, Easington and Stockton currently operate a computerised diabetes register which has to date produced improvements in the quality of care but performance has now plateaued leaving substantial scope for further improvement. This study will evaluate the effectiveness and efficiency of an area wide 'extended' system incorporating a full structured recall and management system, actively involving patients and including clinical management prompts to primary care clinicians based on locally-adapted evidence based guidelines. METHODS: The study design is a two-armed cluster randomised controlled trial of 61 practices incorporating evaluations of the effectiveness of the system, its economic impact and its impact on patient wellbeing and functioning. PMID- 11914162 TI - Organization specific predictors of job satisfaction: findings from a Canadian multi-site quality of work life cross-sectional survey. AB - BACKGROUND: Organizational features can affect how staff view their quality of work life. Determining staff perceptions about quality of work life is an important consideration for employers interested in improving employee job satisfaction. The purpose of this study was to identify organization specific predictors of job satisfaction within a health care system that consisted of six independent health care organizations. METHODS: 5,486 full, part and causal time (non-physician) staff on active payroll within six organizations (2 community hospitals, 1 community hospital/long-term care facility, 1 long-term care facility, 1 tertiary care/community health centre, and 1 visiting nursing agency) located in five communities in Central West Ontario, Canada were asked to complete a 65-item quality of work life survey. The self-administered questionnaires collected staff perceptions of: co-worker and supervisor support; teamwork and communication; job demands and decision authority; organization characteristics; patient/resident care; compensation and benefits; staff training and development; and impressions of the organization. Socio-demographic data were also collected. RESULTS: Depending on the organization, between 15 and 30 (of the 40 potential predictor) variables were found to be statistically associated with job satisfaction (univariate analyses). Logistic regression analyses identified the best predictors of job satisfaction and these are presented for each of the six organizations and for all organizations combined. CONCLUSIONS: The findings indicate that job satisfaction is a multidimensional construct and although there appear to be some commonalities across organizations, some predictors of job satisfaction appear to be organization and context specific. PMID- 11914165 TI - High-tech and low-tech orthopaedic surgery in Sub-Saharan Africa. AB - BACKGROUND: Zambia's governmental health system suffers from shortage of surgical supplies and poor management skills for the sparse resources at hand. The situation has been worsened by the dual epidemics of HIV disease and tuberculosis. On the other hand the private medical sector has benefited greatly from less bureaucracy under the goverment of the Movement for Multi-party Democracy. DISCUSSION: The Zambian-Italian Orthopaedic Hospital in Lusaka is a well organized small unit providing free treatment of physically disabled children. The running costs are met from the fees charged for private consultations, supplemented by donations. State of the art surgical techniques are being used for congenital and acquired musculo-skeletal abnormalities. Last year 513 patients were operated upon free of charge and 320 operations were performed on private patients. PMID- 11914164 TI - Inter-rater agreement in the scoring of abstracts submitted to a primary care research conference. AB - BACKGROUND: Checklists for peer review aim to guide referees when assessing the quality of papers, but little evidence exists on the extent to which referees agree when evaluating the same paper. The aim of this study was to investigate agreement on dimensions of a checklist between two referees when evaluating abstracts submitted for a primary care conference. METHODS: Anonymised abstracts were scored using a structured assessment comprising seven categories. Between one (poor) and four (excellent) marks were awarded for each category, giving a maximum possible score of 28 marks. Every abstract was assessed independently by two referees and agreement measured using intraclass correlation coefficients. Mean total scores of abstracts accepted and rejected for the meeting were compared using an unpaired t test. RESULTS: Of 52 abstracts, agreement between reviewers was greater for three components relating to study design (adjusted intraclass correlation coefficients 0.40 to 0.45) compared to four components relating to more subjective elements such as the importance of the study and likelihood of provoking discussion (0.01 to 0.25). Mean score for accepted abstracts was significantly greater than those that were rejected (17.4 versus 14.6, 95% CI for difference 1.3 to 4.1, p = 0.0003). CONCLUSIONS: The findings suggest that inclusion of subjective components in a review checklist may result in greater disagreement between reviewers. However in terms of overall quality scores, abstracts accepted for the meeting were rated significantly higher than those that were rejected. PMID- 11914166 TI - Scientific evidence in psychiatry: some cautionary notes. PMID- 11914163 TI - Outcomes research in the development and evaluation of practice guidelines. AB - BACKGROUND: Practice guidelines have been developed in response to the observation that variations exist in clinical medicine that are not related to variations in the clinical presentation and severity of the disease. Despite their widespread use, however, practice guideline evaluation lacks a rigorous scientific methodology to support its development and application. DISCUSSION: Firstly, we review the major epidemiological foundations of practice guideline development. Secondly, we propose a chronic disease epidemiological model in which practice patterns are viewed as the exposure and outcomes of interest such as quality or cost are viewed as the disease. Sources of selection, information, confounding and temporal trend bias are identified and discussed. SUMMARY: The proposed methodological framework for outcomes research to evaluate practice guidelines reflects the selection, information and confounding biases inherent in its observational nature which must be accounted for in both the design and the analysis phases of any outcomes research study. PMID- 11914167 TI - Amphetamine salt compound treatment for adults with attention deficit hyperactivity disorder. PMID- 11914168 TI - Assessment and treatment of depression in medically ill children. AB - The consequences of depression in medically ill children include the disability and morbidity that is associated with depression in any patient. They also include an exacerbation of the underlying medical disease and nonadherence to treatment. Thus, medically ill children who are depressed constitute a special, high-risk group of patients who may suffer from severe consequences above and beyond those that are expected in a medically sound population. This manuscript reviews methodologic and practical difficulties that are associated with the diagnosis and treatment of depression in this group of patients. Because treatment of depression in medically ill children may well lead to improvement in medical and psychiatric outcome, there is pressing need for the careful study of potential treatment options in this specific group of patients. PMID- 11914169 TI - Evidence-based treatment for mental disorders in children and adolescents. AB - In the past decade, increased emphasis has been placed on identifying treatments for childhood disorders that are supported by empirical evidence of their effectiveness. This process was spearheaded by an American Psychological Association division 12 task force that identified evidence-based treatments- mostly for disorders of adulthood. Because of the publication of the task force results, other studies have been published that contribute to the knowledge base of evidence-based treatment, and these studies are briefly reviewed. Across evidence-based treatments, common features of effective treatments, such as parent involvement, use of a treatment manual, and the emphasis on generalization of treatment effects to natural settings, are also identified and reviewed.Introduction PMID- 11914170 TI - The Surgeon General's National Action Agenda on Children's Mental Health. AB - The Surgeon General's National Action Agenda highlighted key issues that challenge the public health system in appropriately meeting the mental health needs of children and their families. Among these issues included the need for screening and early identification, improving access to appropriate mental health care, strengthening the infrastructure, and expanding training for providers. Two key gate-keeping systems identified as critical in this reform of mental health care for children and their families are education and primary care. This paper focuses on these two systems to illustrate problems related to these issues. Central to the Surgeon General's call for reform is also the pressing public health responsibility to improve and use the science base by strengthening the connection between what we know from the scientific evidence base and what we do in practice. The implications of this reform for specialty mental health are discussed. PMID- 11914171 TI - Diagnosis and treatment of body dysmorphic disorder in adolescents. AB - Body dysmorphic disorder (BDD) is an underrecognized and underdiagnosed problem that is relatively common among adolescents. Although the onset of the disorder occurs in adolescence, BDD research in child and adolescent psychiatry is relatively limited. Body dysmorphic disorder has a high rate of co-morbidity with depression and suicide, which indicates important implications for prompt diagnosis and treatment in adolescents with BDD. Effective treatment options include cognitive behavioral therapy (CBT) and pharmacotherapy with serotonin reuptake inhibitors (SRIs). This paper provides a brief overview of BDD in adolescents, presents and evaluates the most recent literature on approaches to diagnosis and treatment, and highlights some of the characteristics that distinguish BDD from other disorders such as obsessive-compulsive disorder, social phobia, depression, and eating disorders. PMID- 11914172 TI - Pharmacogenetics of bipolar disorder. AB - To review the pharmacogenetics of bipolar disorders, the authors searched databases for genetic association and linkage studies involving response to long term prophylactic lithium treatment, as well as treatment with antidepressants or clozapine. Significant ethnic variations in the metabolism and efficacy of antidepressants, as well as clozapine, have been reported by several groups. Systematic studies suggest that that genetic factors affect the response to prophylactic lithium treatment. Numerous associations between the three traits of interest and candidate gene polymorphisms have been proposed. Among these, an association between the serotonin transporter gene and response to serotonin reuptake inhibitors appears robust. Considerable interest has also focused on serotonergic gene polymorphisms and response to clozapine. Response to pharmacotherapy in bipolar disorders may be mediated by genetic factors, but the role played by heritability is unknown. PMID- 11914173 TI - Review of bipolar molecular linkage and association studies. AB - This paper reviews the history of molecular genetic linkage and linkage disequilibrium (LD) or association studies of bipolar disorder (BPD). The topic is introduced with a brief discussion of various genetic concepts, including linkage and linkage disequilibrium. It is emphasized that criteria for declaring linkage must include independent confirmation by multiple groups of investigators. Given that the inherited susceptibility for BPD is most likely explained by multiple genes of small effect, simulations indicate that universal confirmation of valid linkages cannot be expected due to sampling variation and genetic heterogeneity. With this background, several valid linkages of BPD to genomic regions are reviewed, including some which may be shared with schizophrenia. These results suggest that nosology must be changed to reflect the genetic origins of the multiple disorders which are collectively described by the term, BPD. The history of BPD LD studies is reviewed, using monoamine oxidase as as an example. Some suggestions of improving these BPD LD are offered. PMID- 11914176 TI - Genetics of early onset bipolar affective disorder: are we making progress? AB - Though considerable progress has been made in understanding the molecular genetics of bipolar affective dis order, few studies are currently focusing on the genetics of prepubertal or early adolescent onset illness. A variety of studies of the phenomenology, imaging and comorbidity of early onset bipolarity find significant differences from late onset illness. These studies raise the notion that early onset cases may represent a distinct genetic form or forms of manic-depressive illness. PMID- 11914174 TI - Family, twin, and adoption studies of bipolar disease. AB - Bipolar disease features states of severe depression that usually fluctuate with at least one episode of intense elation or mania. It is a disorder that has been thought for some time to have a heritable component. The lifetime prevalence of bipolar disease in the general population is approximately 1%. In contrast, family studies have shown the approximate lifetime risk of a first-degree relative of a bipolar proband to be 5% to 10%. Moreover, studies of monozygotic twins show that their risk of contracting the disease is as much as 75 times greater than that for the general population. In addition, adoption studies have demonstrated that biological relatives of bipolar patients are substantially more likely to have the disorder than are adoptive relatives. PMID- 11914175 TI - Triplet repeats and bipolar disorder. AB - Anticipation, the phenomenon of a disease becoming more severe or having earlier onset as it is transmitted down the generations, was originally described in families with psychiatric illness but was thought due to ascertainment bias and became forgotten. Interest was rekindled when a number of neurodegenerative disorders that show this phenomenon, were found to be due to a novel form of mutation--unstable triplet repeats showing intergenerational expansion. Some recent studies of anticipation are consistent with its occurrence in bipolar disorder but are still associated with methodological problems making interpretation difficult. A number of case-control studies employing the repeat expansion detection (RED) technique have found longer repeats in bipolar probands but other studies have found no such association. Despite a large number of studies examining the role of various repeat containing candidate genes, a pathogenic triplet repeat has yet to be found for bipolar disorder. It is likely that the controversy surrounding anticipation and the existence of triplet repeats will only finally be resolved with the demonstration of such a mutation in the aetiology of bipolar disorder. PMID- 11914177 TI - Familial links between attention deficit hyperactivity disorder, conduct disorder, and bipolar disorder. AB - Although family, twin, and adoption studies indicate that attention deficit hyperactivity disorder (ADHD) is a familial condition with a robust genetic component, molecular genetic studies of candidate genes have produced inconsistent findings. One of the challenges to elucidating the genetic architecture of ADHD is its potential genetic heterogeneity. Therefore, efforts are needed to identify etiologically homogeneous subgroups of subjects with ADHD for use in genetic studies. The current article reviews evidence suggesting that parsing ADHD subjects based on comorbidity with conduct and bipolar disorders may yield familial subtypes that are suitable for genetic analyses. PMID- 11914180 TI - Progress in breast cancer chemoprevention. AB - Over the past years there have been significant advances in breast cancer treatment and early detection. For the first time, a decrease in cancer mortality has been observed. Recently, much progress has been made in the understanding of carcinogenesis partly due to available new technologies to detect early molecular changes in the tissue. The knowledge of breast cancer carcinogenesis has provided possible opportunities to prevent breast cancer. Currently, several clinical breast cancer prevention trials are ongoing. This paper reviews issues related to breast cancer chemoprevention including identification of high risk cohorts, endpoint biomarkers, current and new chemopreventive agents as well as models to evaluate these agents. PMID- 11914179 TI - Estrogen receptor beta in breast cancer. AB - Estrogen is essential for normal growth and differentiation in the mammary gland. It also supports growth of approximately 50% of primary breast cancers. For this reason, removal of estrogen or blocking of its action with the anti-estrogen, tamoxifen, is the main treatment for estrogen receptor alpha (ERalpha)-positive tumors. In 1996, when oncologists became aware of a second ER, ERbeta, there was some doubt as to whether this receptor would be of importance in breast cancer because the clinical consensus was that responsiveness to tamoxifen is related to the presence of ERalpha in breast cancer. Today we know that ERalpha and ERbeta have distinct cellular distributions, regulate separate sets of genes and can oppose each other's actions on some genes. We also know that ERbeta is widely expressed in both the normal and malignant breast and that there are proliferating cells in the breast which express ERbeta. In this review we summarize what is known about ERbeta in breast cancer and examine the possibility that ERbeta-selective ligands may well represent a useful class of pharmacological tools with a novel target, namely proliferating cells expressing ERbeta. PMID- 11914181 TI - Vaccination against the HER-2/neu oncogenic protein. AB - The HER-2/neu oncogenic protein is a well-defined tumor antigen. HER-2/neu is a shared antigen among multiple tumor types. Patients with HER-2/neu protein overexpressing breast, ovarian, non-small cell lung, colon, and prostate cancers have been shown to have a pre-existent immune response to HER-2/neu. No matter what the tumor type, endogenous immunity to HER-2/neu detected in cancer patients demonstrates two predominant characteristics. First, HER-2/neu-specific immune responses are found in only a minority of patients whose tumors overexpress HER 2/neu. Secondly, immunity, if detectable, is of low magnitude. These observations have led to the development of vaccine strategies designed to boost HER-2/neu immunity in a majority of patients. HER-2/neu is a non-mutated self-protein, therefore vaccines must be developed based on immunologic principles focused on circumventing tolerance, a primary mechanism of tumor immune escape. HER-2/neu specific vaccines have been tested in human clinical trials. Early results demonstrate that significant levels of HER-2/neu immunity can be generated with active immunization. The T-cell immunity elicited is durable after vaccinations have ended. Furthermore, despite the generation of CD8(+) and CD4(+) T-cells responsive to HER-2/neu in a majority of patients, there is no evidence of autoimmunity directed against tissues that express basal levels of the protein. Cancer vaccines targeting the HER-2/neu oncogenic protein may be useful adjuvants to standard therapy and aid in the prevention of relapse in patients whose tumors overexpress the protein. Furthermore, boosting HER-2/neu-specific T-cell frequencies via active immunization may allow the ex vivo expansion of HER-2/neu specific T-cells for use in adoptive immunotherapy, a therapeutic strategy directed against the treatment of established disease. PMID- 11914182 TI - Mechanisms implicated in the growth regulatory effects of vitamin D in breast cancer. AB - It is now well established that, in addition to its central role in the maintenance of extracellular calcium levels and bone mineralization, 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), the active form of vitamin D, also acts as a modulator of cell growth and differentiation in a number of cell types, including breast cancer cells. The anti-proliferative effects of 1,25(OH)(2)D(3) have been linked to suppression of growth stimulatory signals and potentiation of growth inhibitory signals, which lead to changes in cell cycle regulators such as p21(WAF-1/CIP1) and p27(kip1), cyclins and retinoblastoma protein as well as induction of apoptosis. Such studies have led to interest in the potential use of 1,25(OH)(2)D(3) in the treatment or prevention of certain cancers. Since this approach is limited by the tendency of 1,25(OH)(2)D(3) to cause hypercalcaemia, synthetic vitamin D analogues have been developed which display separation of the growth regulating effects from calcium mobilizing actions. This review examines mechanisms by which 1,25(OH)(2)D(3) and its active analogues exert both anti proliferative and pro-apoptotic effects and describes some of the synthetic analogues that have been shown to be of particular interest in relation to breast cancer. PMID- 11914185 TI - Ethical issues in human genome epidemiology: a case study based on the Japanese American Family Study in Seattle, Washington. AB - Recent completion of the draft sequence of the human genome has been greeted with both excitement and skepticism, and the potential of this accomplishment for advancing public health has been tempered by ethical concerns about the protection of human subjects. This commentary explores ethical issues arising in human genome epidemiology by using a case study approach based on the ongoing Japanese American Family Study at the University of Washington in Seattle (1994 2003). Ethical issues encountered in designing the study, collecting the data, and reporting the study results are considered. When developing studies, investigators must consider whether to restrict the study to specific racial or ethnic groups and whether community involvement is appropriate. Once the study design is in place, further ethical issues emerge, including obtaining informed consent for DNA banking and protecting the privacy and confidentiality of family members. Finally, investigators must carefully consider whether to report genotype results to study participants and whether pedigrees illustrating the results of the study will be published. Overall, the promise of genomics for improving public health must be pursued based on the fundamental ethical principles of respect for persons, beneficence, and justice. PMID- 11914186 TI - Flexible matching strategies to increase power and efficiency to detect and estimate gene-environment interactions in case-control studies. AB - Lack of power is a pertinent problem in many case-control studies of gene environment interactions. The authors recently introduced the concept of flexible matching strategies with varying proportions of a matching factor among selected controls (degree of matching) to increase the power and efficiency of case control studies. In this study, they extended the concept of flexible matching strategies to the field of gene-environment interactions. They assessed the power and efficiency of such studies to detect and estimate gene-environment interactions under a variety of assumptions regarding the prevalence and effects of the environmental exposure and the genetic susceptibility as well as their association in the population. For each set of parameters, 10,000 case-control studies were simulated using varying degrees of matching. Traditional frequency matching increased the power and precision in most scenarios, but even greater gains were often obtained by increasing the prevalence of the environmental exposure in controls above the one in cases. The authors concluded that flexible matching strategies can increase the power and efficiency of case-control studies to detect and estimate gene-environment interactions compared with traditional frequency matching and therefore might help to alleviate the notorious lack of power of these studies in specific situations. PMID- 11914183 TI - Mechanisms involved in the progression of androgen-independent prostate cancers: it is not only the cancer cell's fault. AB - The acquisition of an androgen-independent phenotype by prostate cancer cells is presently a death sentence for patients. In order to have a realistic chance of changing this outcome, an understanding of what drives the progression to androgen independence is critical. We review here a working hypothesis based on the position that the development of androgen-independent epithelial cells is the result of a series of cellular and molecular events within the whole tissue that culminates in the loss of normal tissue-maintained growth control. This tissue includes the epithelial and stromal cells, the supporting extracellular matrix and circulating hormones. This review discusses the characteristics of these malignant cells, the role of stromal cells involved in growth and the differentiation of epithelial cells, and the role of the extracellular matrix as a mediator of the phenotypes of stromal and epithelial cells. In addition, environmental, neuroendocrine and immune factors that may contribute to disturbance of the fine balance of the epithelial-stromal-extracellular matrix connection are considered. While the goal of many therapeutic approaches to prostate cancer has been androgen ablation or targeting the androgen receptor (AR) of epithelial cells, these therapies become ineffective as the cells progress beyond dependence on androgen for growth control. Twenty years ago Sir David Smithers debated that cancer is the result of loss of tolerance within tissues and the organizational failure of normal growth-control mechanisms. This is precipitated by prolonged or abnormal demands for regeneration or repair, rather than of any inherent disorder peculiar to each of the individual components involved. He wrote "It is not the cell itself that is disorderly, but its relationship with the rest of the tissue". We have gained significantly large amounts of precise data on the effects of androgenic ablation on cancerous prostate cells and on the role of the AR in prostate cancer. The need has come to compile this information towards a perspective of dysregulation of tissue as a whole, and to develop experimental systems to address this broader perspective to find and develop therapies for treatment and prevention. PMID- 11914187 TI - Birth characteristics and leukemia in young children. AB - The relation between birth characteristics and leukemia in young children was investigated in a large population-based study in California. Cases were obtained from the statewide cancer registry for 1988-1997. During this time, 1,957 leukemia cases were diagnosed among children under age 5 years. Of these, 1,728 (88%) were matched to a California birth certificate. Two control birth certificates, matched on date of birth and sex, were randomly selected from the statewide birth registry for each case. Analyses were performed separately for acute lymphoid leukemia (ALL) and acute nonlymphoid leukemia (ANLL). Odds ratios and 95% confidence intervals were estimated from conditional logistic regression. The strongest finding was for greatly increased risk of both types of leukemia in children with Down's syndrome (22 cases and no controls). African-American children had strikingly decreased risk for ALL (odds ratio (OR) = 0.29, 95% confidence interval (CI): 0.20, 0.42), and Asian/Pacific Islanders had increased risk for ANLL (OR = 2.00, 95% CI: 1.19, 3.36). Older maternal age was associated with slightly increased risk for ALL (maternal age > or =35 years, OR = 1.25, 95% CI: 1.04, 1.52), although this odds ratio was somewhat reduced when adjusted for other factors. No strong relations were observed for birth weight and ALL or ANLL. PMID- 11914188 TI - Cutaneous melanin density of Caucasians measured by spectrophotometry and risk of malignant melanoma, basal cell carcinoma, and squamous cell carcinoma of the skin. AB - Recent advances have enabled quite accurate estimation by spectrophotometry of the density of cutaneous melanin. The relation between skin cancers and this objective measure of skin phenotype is examined here. For this purpose, a population-based case-control study of subjects aged 20-59 years of northern European ancestry was conducted in Tasmania, Australia. Cases (n = 244) of cutaneous malignant melanoma during 1998-1999, and a sample of cases of basal cell carcinoma (n = 220) and squamous cell carcinoma (n = 195) of the skin were identified from cancer registrations. Controls (n = 483) were selected from a comprehensive population listing. Melanin at the upper inner arm was estimated from skin reflectance of light of 400 and 420 nm wavelengths. For melanoma, basal cell carcinoma, and squamous cell carcinoma, respectively, the odds ratios comparing the least with the highest of four melanin categories were 6.2 (95% confidence interval (CI): 2.3, 16.6), 6.3 (95% CI: 2.6, 15.1), and 4.2 (95% CI: 1.7, 10.8) for men and 1.9 (95% CI: 1.0, 3.7), 1.4 (95% CI: 0.7, 3.0), and 0.7 (95% CI: 0.3, 1.7) for women. The gender differences were not due to disparities in site of occurrence or (for melanoma) in thickness of the lesion. The authors conclude that, particularly for men, cutaneous melanin density at the upper inner arm is a strong predictor of risk of skin cancer. PMID- 11914189 TI - Is residual confounding a reasonable explanation for the apparent protective effects of beta-carotene found in epidemiologic studies of lung cancer in smokers? AB - The results of three randomized trials of beta-carotene supplementation for the prevention of lung cancer among smokers are in contradiction to a large body of epidemiologic evidence for the reduction of risk of lung cancer among smokers with higher intake and/or higher serum levels of beta-carotene. Complicating this issue are widely noted negative associations between tobacco use and intake or serum levels of beta-carotene. Although observational studies attempt to control for reported smoking histories, the accuracy of self-reported smoking is uncertain; correlations as low as 0.5 between reported and true smoking exposure are not inconsistent with studies of biomarkers of cigarette exposure. The authors developed a simple statistical model for random errors in reported smoking (relative to true tobacco exposure) and assumed a modest (inverse) relation between true tobacco exposure and serum beta-carotene. Calculations from this model, combined with a model for lung cancer contemplated by Doll and Peto (J Epidemiol Community Health 1978;78:303-13), suggest that biases in assessment of smoking exposure between smokers with low versus high beta-carotene intake may plausibly explain much or all of the observed protective effect of high beta carotene levels. Appropriate cohort studies of lung cancer in smokers, utilizing biomarkers of smoking, are needed and are presently ongoing. PMID- 11914190 TI - Plasma concentrations of polychlorinated biphenyls and the risk of breast cancer: a congener-specific analysis. AB - Some reports indicate that exposure to specific polychlorinated biphenyl (PCB) congeners is related to breast cancer risk. The authors recruited participants in a case-control study from October 1994 to March 1997 to assess the relation between breast cancer risk and concentrations of 14 PCB congeners measured in plasma lipids by high-resolution gas chromatography. Participants were incident cases of breast cancer (n = 314) and controls (n = 523) from the Quebec City region (Canada). Compared with controls, cases had significantly higher concentrations of PCB 99 (p = 0.02), PCB 118 (p = 0.03), and PCB 156 (p = 0.006). Associations were found between breast cancer risk and either PCB 118 (odds ratio (OR) = 1.60, 95% confidence interval (CI): 1.01, 2.53; fourth vs. first quartile) or PCB 156 (OR = 1.80, 95% CI: 1.11, 2.94; fourth vs. first quartile) concentration. Breast cancer risk was also associated with a total concentration of the three mono-ortho-substituted congeners 105, 118, and 156 expressed as 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (OR = 2.02, 95% CI: 1.24, 3.28; fourth vs. first quartile). These results suggest that exposure to dioxin like PCBs increases breast cancer risk. Alternatively, the results may be explained by differences between cases and controls regarding metabolic pathways involved in the biotransformation of both mono-ortho PCBs and estrogens. PMID- 11914191 TI - Protein consumption and bone mineral density in the elderly : the Rancho Bernardo Study. AB - The role of dietary protein in osteoporosis is unclear, with previous studies having suggested both protection and harm. The associations of total, animal, and vegetable protein with bone mineral density (BMD) and the variations in these associations with calcium intake were studied in a community-dwelling cohort of 572 women and 388 men aged 55-92 years (Rancho Bernardo, California). Multiple linear regression analyses adjusted for standard osteoporosis covariates showed a positive association between animal protein consumption, assessed by food frequency questionnaires in 1988-1992, and BMD, measured 4 years later. This association was statistically significant in women. For every 15-g/day increase in animal protein intake, BMD increased by 0.016 g/cm2 at the hip (p = 0.005), 0.012 g/cm2 at the femoral neck (p = 0.02), 0.015 g/cm2 at the spine (p = 0.08), and 0.010 g/cm2 for the total body (p = 0.04). Conversely, a negative association between vegetable protein and BMD was observed in both sexes. Some suggestion of effect modification by calcium was seen in women, with increasing protein consumption appearing to be more beneficial for women with lower calcium intakes, but evidence for this interaction was not consistently strong. This study supports a protective role for dietary animal protein in the skeletal health of elderly women. PMID- 11914192 TI - Risk of hepatitis C virus infection among young adult injection drug users who share injection equipment. AB - Designing studies to examine hepatitis C virus (HCV) transmission via the shared use of drug injection paraphernalia other than syringes is difficult because of saturation levels of HCV infection in most samples of injection drug users (IDUs). The authors measured the incidence of HCV infection in a large cohort of young IDUs from Chicago, Illinois, and determined the risk of HCV seroconversion associated with specific forms of sharing injection paraphernalia. From 1997 to 1999, serum samples obtained from 702 IDUs aged 18-30 years were screened for HCV antibodies; prevalence was 27%. Seronegative participants were tested for HCV antibodies at baseline, at 6 months, and at 12 months. During 290 person-years of follow-up, 29 participants seroconverted (incidence: 10.0/100 person-years). The adjusted relative hazard of seroconversion, controlling for demographic and drug use covariates, was highest for sharing "cookers" (relative hazard = 4.1, 95% confidence interval: 1.4, 11.8), followed by sharing cotton filters (relative hazard = 2.4, 95% confidence interval: 1.1, 5.0). Risks associated with syringe sharing and sharing of rinse water were elevated but not significant. After adjustment for syringe-sharing, sharing cookers remained the strongest predictor of seroconversion (relative hazard = 3.5, 95% confidence interval: 1.3, 9.9). The authors conclude that sharing of injection equipment other than syringes may be an important cause of HCV transmission between IDUs. PMID- 11914193 TI - Determination of the prevalence of infection with Mycobacterium tuberculosis among persons vaccinated against Bacillus Calmette-Guerin in South Korea. AB - The prevalence of tuberculous infection was estimated among 12,032 persons with a Bacillus Calmette-Guerin (BCG) vaccination scar and 7,788 persons without such a scar who participated in a nationwide tuberculin skin test survey conducted in the Republic of Korea in 1975. The analysis was built upon mixture models that captured the heterogeneity of indurations arising from tuberculous infection, cross-reactions due to infection with environmental mycobacteria, and BCG vaccination. The three distributions were allowed to vary by age, sex, and BCG vaccination status in the Bayesian manner, according to the prior opinion of the authors. Estimated prevalences of tuberculous infection were similar among persons with a BCG scar and persons without one: 7.5% (95% credibility interval (CI): 3.1, 12.5) and 5.2% (95% CI: 4.2, 6.3), respectively, at age 0-4 years and 87.3% (95% CI: 84.0, 90.2) and 84.0% (95% CI: 81.9, 85.8), respectively, at age 25-29 years. From this analysis it can be concluded that mixture models allow investigators, for the first time, to estimate the prevalence of tuberculous infection not only in unvaccinated persons but also in the BCG-vaccinated population. Mixture models are a versatile tool for analyzing diagnostic test data and more general classification problems of considerable complexity. PMID- 11914194 TI - Epidemiologic studies of human semen quality: considerations for study design. AB - Few empirical data exist on the characteristics of subjects who provide semen specimens in epidemiologic studies. The objective of this investigation was to determine participation rates and potential biases in a contemporary study of human semen quality. Subjects (n = 268) are a subset of the Child Health and Development Studies. Their mothers enrolled between 1960 and 1963 during pregnancy. Archived prenatal serum samples, prenatal and birth records, placental examinations, and follow-up for growth and development through adulthood are available. Sons were aged 36-39 years at the time of this study. Respondents to the initial mailing and nonrespondents, who were subsequently traced and recruited, differed in semen parameters, including sperm concentration (78.3 x 10(6)/ml for respondents vs. 37.2; p = 0.003) and the percentage of normal morphology according to the 1987 criteria of the World Health Organization (58.7% for respondents vs. 53.3%; p = 0.04). The authors conclude that researchers designing population-based studies of semen parameters should expect nonrepresentative samples. Adaptation of the design to anticipate and mitigate bias and to maximize efficiency can yield scientifically sound information. Recommendations for study designs are discussed. PMID- 11914195 TI - Recall of early menstrual history and menarcheal body size: after 30 years, how well do women remember? AB - The validity of recall of early menstrual characteristics is of interest because of their putative role in the etiology of breast cancer and other diseases. A retrospective follow-up of the Newton Girls Study (1965-1975) provided an opportunity to assess the accuracy and precision of recall of several early menstrual characteristics. In 1998-1999, 57 percent of the original 793 Newton Girls Study participants completed a mailed questionnaire to assess the accuracy of recall for age and body size at menarche, usual cycle length during the first 2 years, and age at regularity. Recalled and original age at menarche were highly correlated (r = 0.79, p < 0.001). The body mass index percentile at menarche was well correlated with recalled body size at menarche (r = 0.61, p < 0.001), but with some evidence of systematic bias. Overall, a woman's recall of menarcheal age and body size was better than recall of cycle length and occurrence of regularity. The failure to identify certain menstrual characteristics as exposures for subsequent disease may reflect limitations in the accuracy and precision of the recalled measures. PMID- 11914197 TI - Is diabetic retinopathy an inflammatory disease? PMID- 11914198 TI - Blood pressure control and diabetic retinopathy. PMID- 11914199 TI - Anti-angiogenic therapy for uveal melanoma--more haste, less speed. PMID- 11914200 TI - I went; I saw: I was never the same! PMID- 11914202 TI - Periorbital dermatitis as a side effect of topical dorzolamide. AB - AIM: To report periorbital dermatitis as a late side effect of topical dorzolamide hydrochloride (Trusopt), a drug used to reduce intraocular pressure. METHODS: A retrospective study of 14 patients who developed periorbital dermatitis while using topical dorzolamide hydrochloride was undertaken. Six patients underwent patch testing for sensitivity to Trusopt, dorzolamide hydrochloride, and the preservative benzalkonium chloride. RESULTS: The periorbital dermatitis occurred after a mean period of 20.4 weeks of commencing dorzolamide hydrochloride therapy. 13 patients had used preserved topical beta blocker treatment for a mean period of 34.2 months without complication before the introduction of dorzolamide. In eight (57.1%) the dermatitis resolved completely after discontinuing dorzolamide but in six (42.9%) resolution of the dermatitis did not occur until the concomitant preserved beta blocker was stopped and substituted with preservative free drops. Patch testing for sensitivity to Trusopt, dorzolamide hydrochloride, and benzalkonium chloride was negative. CONCLUSION: These findings suggest that dorzolamide can cause severe periorbital dermatitis. Although the dermatitis may resolve when dorzolamide is discontinued, this does not always occur and in some patients all topical medication containing benzalkonium chloride needs to be stopped. PMID- 11914201 TI - Moderate visual impairment in India: the Andhra Pradesh Eye Disease Study. AB - AIM: To assess the prevalence and demographic associations of moderate visual impairment in the population of the southern Indian state of Andhra Pradesh. METHODS: From 94 clusters in one urban and three rural areas of Andhra Pradesh, 11 786 people of all ages were sampled using a stratified, random, cluster, systematic sampling strategy. The eligible people were invited for interview and detailed dilated eye examination by trained professionals. Moderate visual impairment was defined as presenting distance visual acuity less than 6/18 to 6/60 or equivalent visual field loss in the better eye. RESULTS: Of those sampled, 10,293 (87.3%) people participated in the study. In addition to the previously reported 1.84% prevalence of blindness (presenting distance visual acuity less than 6/60 or central visual field less than 20 degrees in the better eye) in this sample, 1237 people had moderate visual impairment, an adjusted prevalence of 8.09% (95% CI 6.89 to 9.30%). The majority of this moderate visual impairment was caused by refractive error (45.8%) and cataract (39.9%). Increasing age, female sex, decreasing socioeconomic status, and rural area of residence had significantly higher odds of being associated with moderate visual impairment. CONCLUSIONS: These data suggest that there is a significant burden of moderate visual impairment in this population in addition to blindness. Extrapolation of these data to the population of India suggests that there were 82 million people with moderate visual impairment in the year 2000, and this number is likely to be 139 million by the year 2020 if the current trend continues. This impending large burden of moderate visual impairment, the majority of which is due to the relatively easily treatable refractive error and cataract, would have to be taken into account while estimating the eye care needs in India, in addition to dealing with blindness. Specific strategies targeting the elderly population, people with low socioeconomic status, those living in the rural areas, and females would have to be implemented in the long term to reduce moderate visual impairment. PMID- 11914203 TI - Pathology of cyclodiode laser: a series of nine enucleated eyes. AB - AIM: To study the histological effects of cyclodiode laser treatment in humans, and to compare these findings with the clinical course, treatment response, complications, and indications for enucleation. METHOD: Detailed histological examination of nine enucleation specimens was undertaken in conjunction with a retrospective review of patient case notes. RESULTS: Retreatments had been undertaken in three cases. Although all globes showed damage to pars plicata, intact ciliary processes within the treatment zone were present in all cases. Pars plana injury was also noted in two thirds of cases. Inflammation was mild. Ciliary epithelial proliferation was seen in most cases with increasing time following treatment, in a disorganised pattern, without replication of the ciliary epithelial bilayer. No regeneration of the ciliary processes with fibrovascular cores was found. The three patients with good IOP control at enucleation had all had multiple diode treatments. Neither phthisis nor sympathetic ophthalmia was seen. CONCLUSIONS: Diode laser cyclophotocoagulation produces very characteristic injury to pars plicata, which frequently extends into pars plana, but with only mild persisting inflammation. Ciliary processes are, however, frequently spared within the treatment zone and may account for early or late treatment failure. PMID- 11914204 TI - Vitreous penetration of levofloxacin in the uninflamed phakic human eye. AB - AIMS: To assess the vitreous penetration of oral levofloxacin (a new fluoroquinolone antibiotic with improved Gram positive activity) in uninflamed phakic eyes. METHODS: 15 patients for macula hole surgery were recruited to the study. 10 received a single 500 mg dose of levofloxacin by mouth preoperatively. Five acted as controls. Serum and undiluted vitreous samples were obtained at surgery and analysed by HPLC. RESULTS: Levofloxacin was detectable 2.5 hours after administration in the vitreous. A peak concentration of 1.6 microg/ml (or mg/l) was measured between 2.5 and 4 hours post-dose. CONCLUSION: Oral levofloxacin reaches the vitreous rapidly in the uninflamed phakic eye. Levels did not reach MIC(90) for the commonest infecting organisms. Nevertheless, levofloxacin would be expected to be active against a higher proportion of infecting organisms than either ciprofloxacin or ofloxacin. PMID- 11914205 TI - Macular hole size as a prognostic factor in macular hole surgery. AB - BACKGROUND/AIM: In 1991 there was a series of successful closures of a macular hole after vitrectomy and membrane peeling. Today this technique has become a standard procedure. The aim of this study was to evaluate the role of optical coherence tomography in diagnosing and staging, as well as in predicting, the functional and anatomical outcome after macular hole surgery. METHOD: In a prospective study 94 consecutive patients (20 male, 74 female) with a mean age of 67.6 (SD 6.0) years and a macular hole stage II (n = 8), III (n = 72), and IV (n = 14) according to the classification by Gass were examined with optical coherence tomography (OCT) before pars plana vitrectomy. Macular hole diameters were determined at the level of the retinal pigment epithelium (base diameter) and at the minimal extent of the hole (minimum diameter). Calculated hole form factor (HFF) was correlated with the postoperative anatomical success rate and best corrected visual acuity. The duration of symptoms was correlated with base and minimum diameter of the macular hole. RESULTS: In eyes without anatomical closure of the macular hole after one surgical approach (13/94) the base diameter (p1) and the minimum diameter (p2) were significantly larger than in cases with immediate postsurgical closure (p1 = 0.003; p2 = 0.028). There was a significant negative correlation between both the base and the minimum diameter of the hole and the postoperative visual function (p1 = 0.016; p2 = 0.002). In all patients with HFF >0.9 the macular hole was closed following one surgical procedure, whereas in eyes with HFF <0.5 anatomical success rate was 67%. Better postoperative visual outcome correlated with higher HFF (p = 0.050). There was no significant correlation between the duration of symptoms and base or minimum diameters (p1 = 0.053; p2 = 0.164), respectively. CONCLUSION: Preoperative measurement of macular hole size with OCT can provide a prognostic factor for postoperative visual outcome and anatomical success rate of macular hole surgery. The duration of symptoms did not correlate with the diameters measured. Base and minimum diameters especially seem to be of predictive value in macular hole surgery. PMID- 11914206 TI - Complications of motility peg placement for porous hydroxyapatite orbital implants. AB - AIM: To evaluate the complications associated with pegging of porous hydroxyapatite orbital implants. METHODS: Complications associated with pegging were retrospectively reviewed from the charts of 100 of 133 patients with hydroxyapatite implantation from 1993 to 2000. RESULTS: 48 (48%) of the 100 hydroxyapatite implanted patients who had undergone pegging were found to have problems with their pegs, including discharge (45.8%), peg falling out (20.8%), pyogenic granulomas (16.7%), popping peg (14.6%), hydroxyapatite visible around peg hole (8.3%), part of peg shaft visible (6.2%), peg drilled off centre (6.2%), peg drilled at an angle (4.2%), and excess movement of peg (4.2%). The standard peg fell out statistically more often than the peg and sleeve system (Yates's corrected chi(2), p=0.038). There was a trend towards complications of the peg with use of a standard peg (versus sleeved peg) (p=0.226). CONCLUSIONS: There are several potential complications of pegging. Most complications are minor and can be managed successfully. PMID- 11914207 TI - Corneal tattooing: an alternative treatment for disfiguring corneal scars. AB - BACKGROUND: The performance and results of corneal tattooing are described in a case series of 11 patients suffering from a disfiguring corneal scar using a technique similar to conventional dermatography. METHODS: Drawing ink in different shades was applied into the anterior corneal stroma by punctures performed with a conventional spatula needle. RESULTS: Up to 4 years after surgery all patients still had satisfactory staining of the formerly cosmetically disfiguring corneal scar. CONCLUSION: Tattooing of unsightly corneal scars proved to be an efficient and easy to perform technique, yielding acceptable results during follow up. PMID- 11914208 TI - A comparative assessment of endothelium from pseudophakic and phakic donor corneas stored in organ culture. AB - AIMS: To evaluate the endothelial quality of corneas obtained from pseudophakic donors and to compare the data with matched phakic controls. METHODS: Corneas from eyes with posterior chamber intraocular lenses (PCIOLs) and corneas from phakic eyes (controls) were stored for 1-2 weeks in organ culture and then examined after staining with Alizarin red S. The corneas were divided into two groups according to the duration of storage. Endothelial cell density, the percentage of hexagonal cells, and the coefficient of variation (CV) were determined. RESULTS: There was no statistically significant difference between the 14 PCIOL corneas and the 13 controls stored in organ culture for 7 days for any of the three parameters studied. The mean cell density was 2155 (SD 529) cells/mm(2) in the PCIOL corneas and 2118 (453) cells/mm(2) in the controls (p=0.85). The mean percentage of hexagonal cells was 52% (8%) and 58% (7%), respectively (p=0.06). The mean CV was 0.32 (0.18) in the pseudophakic corneas and 0.39 (0.18) in the controls (p=0.33). Moreover, there was no significant difference between the PCIOL corneas and the controls stored for up to 2 weeks. CONCLUSIONS: The corneal endothelium from eyes with PCIOLs appears to be similar to that of phakic eyes after 1-2 weeks in organ culture. This finding suggests that corneas from pseudophakic eyes should not routinely be disqualified for transplantation. The use of at least some pseudophakic corneas may substantially increase the potential donor pool. PMID- 11914209 TI - Cornea procurement from very old donors: post organ culture cornea outcome and recipient graft outcome. AB - AIM: To study the suitability of corneas from very old donors for graft after banking and their clinical and endothelial outcomes in recipients. METHODS: 419 corneas stored in organ culture were divided into group 1, donors under 85 years (330 corneas) and group 2, "very old" donors aged 85 years and over (89 corneas). Endothelial cell density (ECD) before and after organ culture, discard rate before and after storage, and clinical and endothelial outcomes of the 196 penetrating keratoplasties (PKP) (158 in group 1 and 38 in group 2) were compared in a prospective longitudinal study. RESULTS: Initial ECD was lower in group 2 than in group 1 and elimination for low ECD was more frequent in group 2 (respectively 38% v 20.2%, p=0.001). At the end of storage, because very old corneas lost fewer ECs than younger ones (respectively 4.2% v 9.5%, p=0.022), ECD was comparable between the two groups. The corneas of very old donors had a poorer macroscopic appearance at procurement and during surgery. Despite this, in grafted patients, overall graft survival in groups 1 and 2 (respectively 87.4% v 80.6%, p=0.197), visual acuity, and ECD did not differ at completion of the study (mean follow up 25 months). CONCLUSION: This study suggests that endothelial cell count during banking ensures that functional and cellular results of PKPs are not dramatically influenced by very old donor age. Considering Europe's ageing population, the very elderly should not be deemed off limits for corneal procurement. PMID- 11914210 TI - Systemic inflammation and innate immune response in patients with previous anterior uveitis. AB - AIM: To determine the presence of systemic inflammation and innate immune responsiveness of patients with a history of acute anterior uveitis but no signs of ocular inflammation at the time of recruitment. METHODS: Tumour necrosis factor alpha (TNF-alpha) production in response to bacterial lipopolysaccharide (LPS) was studied using whole blood culture assay; levels of TNF-alpha in culture supernatants, and soluble interleukin 2 receptor (sIL-2R) in serum were determined by chemiluminescent immunoassay (Immulite); monocyte surface expression of CD11b, CD14, and CD16 and the proportion of monocyte subsets CD14(bright)CD16(-) and CD14(dim)CD16(+) were studied with three colour whole blood flow cytometry; and serum C reactive protein (CRP) levels were determined using immunonephelometric high sensitivity CRP assay. RESULTS: The CRP level (median, interquartile range) was significantly higher in 56 patients with previous uveitis than in 37 controls (1.59 (0.63 to 3.47) microg/ml v 0.81 (0.32 to 2.09) microg/ml; p=0.008). The TNF-alpha concentration of the culture media per 10(5) monocytes was significantly higher in the patient group than in the control group in the presence of LPS 10 ng/ml (1473 (1193 to 2024) pg/ml v 1320 (935 to 1555) pg/ml; p=0.012) and LPS 1000 ng/ml (3280 (2709 to 4418) pg/ml v 2910 (2313 to 3358) pg/ml; p=0.011). The background TNF-alpha release into the culture media was low in both groups. CD14 expression of CD14(bright)CD16(-) monocytes, defined as antibody binding capacity (ABC), was similar for the patients and controls (22,839 (21,038 to 26,020) ABC v 21,657 (19,854 to 25,646) ABC). CONCLUSIONS: Patients with previous acute anterior uveitis show high innate immune responsiveness that may play a part in the development of ocular inflammation. PMID- 11914211 TI - Prevalence of ocular symptoms and signs with preserved and preservative free glaucoma medication. AB - AIM: To determine the incidence of ocular toxicity of preservatives with glaucoma medications. METHODS: A prospective epidemiological survey was carried out in 1999 by 249 ophthalmologists on 4107 patients. Ocular symptoms, conjunctiva, cornea, and eyelids were assessed. A chi(2) test was used for differences between preserved eye drops (P) and preservative free eye drops (PF). RESULTS: 84% patients used P, 13% received PF, and 3% a combination of P and PF eye drops. All symptoms were more prevalent with P than with PF drops (p<0.001): discomfort upon instillation (43% versus 17%), and symptoms between instillations such as burning stinging (40% versus 22%), foreign body sensation (31% versus 14%), dry eye sensation (23% versus 14%), tearing (21% versus 14%), and eyelid itching (18% versus 10%). An increased incidence (>2 times) of ocular signs was seen with P eye drops. The prevalence of signs and symptoms was dose dependent, increasing with the number of P drops. A reduction in the symptoms and signs was observed when patients changed from P to PF eye drops (p<0.001). CONCLUSIONS: Symptoms and signs are less prevalent when PF drops are used. Moreover, most of the adverse reactions induced by P glaucoma medication are reversible after removing preservatives. PMID- 11914212 TI - Increased mortality in women with cataract: a population based follow up of the North London Eye Study. AB - BACKGROUND/AIMS: In diabetics, cataract is associated with higher risk of death. In non-diabetics the data are conflicting, but some indicate an association between one type of cataract (nuclear) and increased mortality. The aim of this study was to estimate and compare age and sex specific mortality for elderly people with and without cataract in a population based cohort. METHODS: A random sample drawn from a defined population of elderly people (age 65 and older) registered with 17 general practice groups in north London formed the study cohort and were followed up for 4 years. The age and sex specific mortality from various causes was estimated and compared in those with and without cataract. RESULTS: In non-diabetics (n=1318), cataract (lens opacity at baseline) was significantly associated with higher mortality in women. The age standardised death rate per 1000 was 39.8 and 24.8 in women with and without cataract, respectively (age adjusted hazard ratio 1.7, confidence limits 1.1 to 2.7, p=0.032). This was not the case in non-diabetic men (hazard ratio 0.9, confidence limits 0.6 to 1.5, p=0.782). The excess mortality in women with cataract was consistent for cardiovascular, respiratory, and other non-cancer causes of death. There was no association between cataract and mortality from cancer. CONCLUSIONS: This study has shown, for the first time, that cataract is associated with higher mortality in women but not in men, among the non-diabetic population. This sex effect suggests that women may be exposed to risk factors that increase both the risk of cataract and mortality, and that men may have little or no exposure to these "sex specific" factors. Possible risk factors that warrant further investigation may be those associated with some pregnancy and childbearing experience. PMID- 11914213 TI - Capillary density and retinal diameter measurements and their impact on altered retinal circulation in glaucoma: a digital fluorescein angiographic study. AB - AIM: Normal pressure glaucoma (NPG) patients exhibit prolonged retinal arteriovenous passage times in fluorescein angiography and colour Doppler imaging suggests increased resistance downstream from the central retinal and posterior ciliary arteries. The aim of the study was to elucidate the morphological source of decreased perfusion and increased resistance of the ocular circulation in NPG. METHODS: Retinal arteriovenous passage time (AVP) and peripapillary arterial and venous diameters were measured in digital scanning laser fluorescein angiograms. For estimation of retinal capillary density the area of the foveal avascular zone (FAZ) and the perifoveal intercapillary area (PIA) was quantified. 36 patients with NPG (mean age 57 (SD 13) years) and 21 healthy subjects (mean age 51 (13) years) were enrolled in the comparative study. RESULTS: In NPG patients the AVP (2.55 (1.1) seconds) was significantly prolonged (p<0.001) when compared with healthy subject data (AVP: 1.70 (0.39) seconds). No differences for arterial or venous diameter, FAZ, and PIA were observed in NPG patients compared with healthy subjects. FAZ, PIA, arterial and venous diameter were not correlated with visual field indices (except venous diameter with PSD, r=0.35 (p<0.05)) or cup to disc ratios. AVP was significantly correlated (p<0.05) with the size of the optic nerve head (r=-0.28), visual field indices (MD: r=-0.3; PSD: r=0.3; CPSD: r=0.3), and contrast sensitivity (r=-0.34). CONCLUSION: AVP times are significantly prolonged in NPG. The slowing of the retinal transit does not result from capillary dropout, or changes of peripapillary arterial or venous diameters with increased vascular resistance. PMID- 11914214 TI - Limited macular translocation with scleral retraction suture. AB - BACKGROUND/AIMS: Macular translocation with scleral imbrication is a new technique for treating subfoveal choroidal neovascular membranes (CNV). This procedure shortens the sclera but may result in a minimal decrease in the internal circumference of the globe and limits the amount of foveal displacement. The authors propose a new scleral retraction suture aimed at decreasing the internal circumference of the globe in an effort to increase foveal displacement. METHODS: Using a cadaver model, they compared the amount of scleral shortening using a standard scleral imbrication technique and a modified three suture scleral retraction technique. Sections of the globes were digitised and specialised software was used to estimate the amount of scleral shortening. Three patients with subfoveal choroidal neovascularisation underwent limited macular translocation using pars plana vitrectomy and macular detachment with the modified scleral suture technique. The main outcome measures were visual acuity, foveal displacement, and complications. RESULTS: In the cadaver model, the scleral retraction suture resulted in a flatter internal scleral fold compared to the standard suture technique and created approximately 890 microm of effective scleral shortening. In the patients who underwent macular translocation and laser photocoagulation of the CNV, visual acuity improved in two patients and worsened in one patient. The range of foveal displacement was 1400-2400 microm. CONCLUSION: The foveal displacements achieved in this limited study compared to median displacement previously published using standard suture techniques demonstrates that the scleral retraction suture technique may be a useful adjunct to limited macular translocation. The advantage of this type of suture in conjunction with translocation may depend on the effective scleral shortening offered by this retraction suture. PMID- 11914215 TI - Uveal melanomas express vascular endothelial growth factor and basic fibroblast growth factor and support endothelial cell growth. AB - BACKGROUND: Tumour microvascularity is a significant determinant of prognosis for a large number of different tumours, including uveal melanoma. The development of blood vessels within these and other tumours is partly controlled by soluble pro angiogenic cytokines, of which basic fibroblast growth factor (bFGF) and vascular endothelial growth factor-A (VEGF) are the best described. METHODS: Because VEGF has been inconsistently found within uveal melanomas and bFGF is described as an autocrine growth factor in cutaneous melanoma, the authors looked at the expression of these cytokines in uveal melanomas using immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). The cross talk between uveal melanoma cells and endothelial cells was then assessed in an in vitro co culture model. RESULTS: While most tumour cells expressed bFGF at the protein level by immunohistochemistry (89%), relatively few (22%) expressed VEGF, and this was of limited extent. All 20 tumours tested by RT-PCR contained mRNA for both bFGF and VEGF. Co-culture experiments using an ATP based bioassay showed that uveal melanomas could support the growth of a rat brain endothelial cell line (GPNT) and human umbilical vein endothelial cells (HUVEC), and that this could be modulated by cytokines and anti-cytokine antibodies. CONCLUSION: These results suggest that angiogenesis within uveal melanoma may be the result of a complex interplay between endothelial and tumour cells, and that bFGF and VEGF could play a part. PMID- 11914216 TI - Vascular endothelial growth factor is elevated in ocular fluids of eyes harbouring uveal melanoma: identification of a potential therapeutic window. AB - BACKGROUND: Improved local treatment of uveal melanoma makes it possible for many patients to retain the affected eye, but a proportion will develop secondary complications such as neovascularisation of the iris (NVI) and require enucleation. Although vascular endothelial growth factor A (VEGF-A) is known to correlate with NVI and can cause NVI in experimental models, this pro-angiogenic cytokine is consistently reported to be absent in uveal melanoma. Novel anti-VEGF therapies are now in clinical trial, and the authors therefore wished to determine whether VEGF-A was indeed elevated in melanoma bearing eyes. METHODS: VEGF-A concentrations were measured in aqueous and vitreous from 19 and 30 enucleated eyes respectively. RESULTS: Elevated VEGF-A concentrations (up to 21.6 ng/ml) were found in melanoma bearing eyes compared with samples from patients undergoing routine cataract extraction (all had values below 0.96 ng/ml). Immunohistochemistry showed VEGF-A protein in the iris and/or ciliary body of 54% and basic fibroblast growth factor (bFGF) in 82% of the eyes examined. VEGF was found to a limited extent and at very low levels in only 9% of these tumours. Aqueous or vitreous VEGF levels showed no apparent correlation with retinal detachment, tumour size, vascularity, or immunohistochemistry. Though limited in number, the highest VEGF levels correlated with previous radiation therapy, and with the presence neovascularisation of the iris or optic nerve head. bFGF was not significantly elevated in ocular fluids: it is known to be a pro-angiogenic agent and was detected in the majority of primary uveal melanomas. CONCLUSION: Based on this study, though the source of VEGF within eyes harbouring uveal melanoma is not clear, these data suggest that anti-VEGF therapy might prove useful in the management of some patients with NVI secondary to uveal melanoma. PMID- 11914217 TI - A reappraisal of cryosurgery for eyelid basal cell carcinomas. AB - BACKGROUND/AIMS: Liquid nitrogen spray freezing has been successfully applied for basal cell carcinomas in the eyelid region, but is not yet in general use. The reasons for this were analysed and the development of a more reliable, safer cryosurgical technique aimed for. METHODS: New cryosurgical apparatus, contact probes with increased freezing power, and a special application technique were developed and clinically tested in a consecutive series of 221 patients with primary basal cell carcinomas of the lid region. Special efforts yielded follow up reports of 220 out of the 221 patients. RESULTS: Experimental measurements and clinical results demonstrated that this cryosurgical technique was at least as effective as spray freezing, with lower risks. The rate of recurrent tumours in patients followed up for 5 years or longer was 5.1% (surgeons first result) respectively 0.6% (result after optimised second cryosurgery). The figures were 6.8%, respectively max 2.7%, when including all patients, independent of follow up time. CONCLUSION: Traditional surgery and histopathology, still used at numerous places, resulted in higher recurrence rates despite extended loss of healthy eyelid tissues and should be abandoned. Micrographic surgery is considered mandatory to save more of the healthy structures and to obtain lower recurrence rates. Cost and time require worldwide restriction of micrographic surgery to selected cases. Updated cryosurgery provides a low cost option to micrographic surgery and results in preservation of eyelid structures and lacrimal pathways, tarsal plate, lid margin. It provides excellent cosmetic results. Thus, primary basal cell carcinomas in the eyelid region of suitable size and location should receive updated cryosurgery, and tumours beyond its range micrographic surgery. PMID- 11914218 TI - A new non-contact optical device for ocular biometry. AB - BACKGROUND: A new commercially available device (IOLMaster, Zeiss Instruments) provides high resolution non-contact measurements of axial length (using partial coherent interferometry), anterior chamber depth, and corneal radius (using image analysis). The study evaluates the validity and repeatability of these measurements and compares the findings with those obtained from instrumentation currently used in clinical practice. METHOD: Measurements were taken on 52 subjects (104 eyes) aged 18-40 years with a range of mean spherical refractive error from +7.0 D to -9.50 D. IOLMaster measurements of anterior chamber depth and axial length were compared with A-scan applanation ultrasonography (Storz Omega) and those for corneal radius with a Javal-Schiotz keratometer (Topcon) and an EyeSys corneal videokeratoscope. RESULTS: Axial length: the difference between IOLMaster and ultrasound measures was insignificant (0.02 (SD 0.32) mm, p = 0.47) with no bias across the range sampled (22.40-27.99 mm). Anterior chamber depth: significantly shorter depths than ultrasound were found with the IOLMaster (-0.06 (0.25) mm, p <0.02) with no bias across the range sampled (2.85-4.40 mm). Corneal radius: IOLMaster measurements matched more closely those of the keratometer than those of the videokeratoscope (mean difference -0.03 v -0.06 mm respectively), but were more variable (95% confidence 0.13 v 0.07 mm). The repeatability of all the above IOLMaster biometric measures was found to be of a high order with no significant bias across the measurement ranges sampled. CONCLUSIONS: The validity and repeatability of measurements provided by the IOLMaster will augment future studies in ocular biometry. PMID- 11914220 TI - Treacher Collins syndrome with novel ophthalmic findings and visceral anomalies. PMID- 11914219 TI - Ex vivo preservation and expansion of human limbal epithelial stem cells on amniotic membrane cultures. AB - BACKGROUND/AIM: Amniotic membrane (AM) transplantation effectively expands the remaining limbal epithelial stem cells in patients with partial limbal stem cell deficiency. The authors investigated whether this action could be produced ex vivo. METHODS: The outgrowth rate on AM was compared among explants derived from human limbus, peripheral cornea, and central cornea. For outgrowth of human limbal epithelial cells (HLEC), cell cycle kinetics were measured by BrdU labelling for 1 or 7 days, of which the latter was also chased in primary cultures, secondary 3T3 fibroblast cultures, and in athymic Balb/c mice following a brief treatment with a phorbol ester. Epithelial morphology was studied by histology and transmission electron microscopy, and phenotype was defined by immunostaining with monoclonal antibodies to keratins and mucins. RESULTS: Outgrowth rate was 0/22 (0%) and 2/24 (8.3%) for central and peripheral corneal explants, respectively, but was 77/80 (96.2%) for limbal explants (p <0.0001). 24 hour BrdU labelling showed a uniformly low (that is, less than 5%) labelling index in 65% of the limbal explants, but a mixed pattern with areas showing a high (that is, more than 40%) labelling index in 35% of limbal explants, and in all (100%) peripheral corneal explants. Continuous BrdU labelling for 7 days detected a high labelling index in 61.5% of the limbal explants with the remainder still retaining a low labelling index. A number of label retaining cells were noted after 7 day labelling followed by 14 days of chase in primary culture or by 21 days of chase after transplantation to 3T3 fibroblast feeder layers. After exposure to phorbol 12-myristate 13-acetate for 24 hours and 7 day labelling, HLEC transplanted in athymic mice still showed a number of label retaining basal cells after 9 days of chase. HLEC cultured on AM were strongly positive for K14 keratin and MUC4 and slightly positive in suprabasal cells for K3 keratin but negative for K12 keratin, AMEM2, and MUC5AC. After subcutaneous implantation in athymic mice, the resultant epithelium was markedly stratified and the basal epithelial cells were strongly positive for K14 keratin, while the suprabasal epithelial cells were strongly positive for K3 keratin and MUC4, and the entire epithelium was negative for K12 keratin and MUC5A/C. CONCLUSIONS: These data support the notion that AM cultures preferentially preserve and expand limbal epithelial stem cells that retain their in vivo properties of slow cycling, label retaining, and undifferentiation. This finding supports the feasibility of ex vivo expansion of limbal epithelial stem cells for treating patients with total limbal stem cell deficiency using a small amount of donor limbal tissue. PMID- 11914221 TI - Optical coherence tomography imaging of severe commotio retinae and associated macular hole. PMID- 11914222 TI - Acute postural drop in optic nerve perfusion after vitrectomy and gas in a patient with diabetic autonomic neuropathy. PMID- 11914223 TI - Acute ocular ischaemia and orbital inflammation associated with systemic lupus erythematosus. PMID- 11914224 TI - Chickenpox neuroretinitis in a 9 year old child. PMID- 11914225 TI - Complication of acupuncture in a patient with Behcet's disease. PMID- 11914226 TI - Bilateral central retinal artery occlusion in Wegener's granulomatosis and alpha(1) antitrypsin deficiency. PMID- 11914227 TI - Corneoscleral fibrous histiocytoma. PMID- 11914228 TI - Haemophilus influenzae corneal ulcer associated with atopic keratoconjunctivitis and herpes simplex keratitis. PMID- 11914229 TI - Should we vaccinate for glaucoma surgery? PMID- 11914230 TI - Blinking and operating: cognition versus vision. PMID- 11914231 TI - The wide field multifocal electroretinogram reveals retinal dysfunction in early retinitis pigmentosa. PMID- 11914232 TI - Day 1 review following cataract surgery: are we seeing the precise details? PMID- 11914233 TI - Choroidal abnormalities in neurofibromatosis type 1 with non-invasive infrared imaging. PMID- 11914234 TI - Corneal melt and perforation secondary to floppy eyelid syndrome in the presence of rheumatoid arthritis. PMID- 11914235 TI - Late opacification of SC60B-OUV acrylic intraocular lenses. PMID- 11914236 TI - Ocular trauma with small framed spectacles. PMID- 11914237 TI - Treatment of superior limbic keratoconjunctivitis with a unilateral bandage contact lens. PMID- 11914238 TI - Keratectasia after PTK. PMID- 11914242 TI - Vitamin C, collagen, and cracks in the plaque. PMID- 11914241 TI - March 26, 2002: rapid access publication. PMID- 11914243 TI - High-density lipoprotein restores endothelial function in hypercholesterolemic men. AB - BACKGROUND: Hypercholesterolemia is a risk factor for atherosclerosis-causing endothelial dysfunction, an early event in the disease process. In contrast, high density lipoprotein (HDL) cholesterol inversely correlates with morbidity and mortality representing a protective effect. Therefore, we investigated the effects of reconstituted HDL on endothelial function in hypercholesterolemic men. METHODS AND RESULTS: Endothelium-dependent and -independent vasodilation to intraarterial acetylcholine and sodium nitroprusside (SNP), respectively, was measured by forearm venous occlusion plethysmography in healthy normo- and hypercholesterolemic men. In hypercholesterolemics, the effects of reconstituted HDL (rHDL; 80 mg/kg IV over 4 hours) on acetylcholine- and SNP-induced changes in forearm blood flow were assessed in the presence or absence of the nitric oxide (NO) synthase inhibitor L-NMMA. Hypercholesterolemics showed reduced vasodilation to acetylcholine but not to SNP compared with normocholesterolemics (P<0.0001). rHDL infusion increased plasma HDL cholesterol from 1.3+/-0.1 to 2.2+/-0.1 mmol/L (P<0.0001, n=18) and significantly enhanced the acetylcholine-induced increase in forearm blood flow without affecting that induced by SNP. rHDL infusion also improved flow-mediated dilation of the brachial artery (to 4.5+/-0.9% from 2.7+/ 0.6%, P=0.02). NO synthase inhibition prevented the improvement in acetylcholine induced vasodilation while leaving the response to SNP unchanged. Albumin infusion in an equivalent protein dose had no effect on vasomotion or lipid levels. CONCLUSIONS: In hypercholesterolemic patients, intravenous rHDL infusion rapidly normalizes endothelium-dependent vasodilation by increasing NO bioavailability. This may in part explain the protective effect of HDL from coronary heart disease and illustrates the potential therapeutic benefit of increasing HDL in patients at risk from atherosclerosis. PMID- 11914244 TI - Strain rate imaging for assessment of regional myocardial function: results from a clinical model of septal ablation. AB - BACKGROUND: Regional myocardial function assessment is essential in the management of coronary artery disease (CAD). Tissue Doppler imaging (TDI) by depicting local myocardial motion can potentially quantify regional myocardial function. Strain rate imaging (SRI) that depicts regional deformation is less susceptible to cardiac translation and tethering and may be superior to TDI for regional function analysis. We examined regional myocardial function using TDI and SRI in a unique clinical model of a small, discrete myocardial infarction. METHODS AND RESULTS: Ten patients with severely symptomatic septal hypertrophy underwent basal septal ablation via intracoronary alcohol injection and had TDI and SRI pre- and postablation. Invasive hemodynamics showed no appreciable change in global function. Peak systolic strain rate was significantly lower postablation versus preablation (-0.5 versus -1.2 s(-1), P<0.001) and when comparing infarct and noninfarct areas (-0.5 versus -1.5 s(-1), P<0.001). In contrast, peak systolic tissue velocities were similar pre- and postablation (3.9 versus 2.9 cm/s, P=0.16) and between infarct and noninfarct areas (2.9 versus 2.2 cm/s, P=0.13). SRI analysis demonstrated reduced systolic function in the peri infarct zone and preserved systolic function in the remote nonischemic zone. CONCLUSION: In the clinical setting of a small, discrete infarct unaccompanied by changes in global function, SRI accurately depicted changes in regional function. These data suggest that SRI may be the optimal method for objective, quantitative assessment of regional myocardial dysfunction. PMID- 11914245 TI - Prevalence of Anderson-Fabry disease in male patients with late onset hypertrophic cardiomyopathy. AB - BACKGROUND: Although studies have suggested that "late-onset" hypertrophic cardiomyopathy (HCM) may be caused by sarcomeric protein gene mutations, the cause of HCM in the majority of patients is unknown. This study determined the prevalence of a potentially treatable cause of hypertrophy, Anderson-Fabry disease, in a HCM referral population. METHODS AND RESULTS: Plasma alpha galactosidase A (alpha-Gal) was measured in 79 men with HCM who were diagnosed at > or =40 years of age (52.9+/-7.7 years; range, 40-71 years) and in 74 men who were diagnosed at <40 years (25.9+/-9.2 years; range, 8-39 years). Five patients (6.3%) with late-onset disease and 1 patient (1.4%) diagnosed at <40 years had low alpha-Gal activity. Of these 6 patients, 3 had angina, 4 were in New York Heart Association class 2, 5 had palpitations, and 2 had a history of syncope. Hypertrophy was concentric in 5 patients and asymmetric in 1 patient. One patient had left ventricular outflow tract obstruction. All patients with low alpha-Gal activity had alpha-Gal gene mutations. CONCLUSION: Anderson-Fabry disease should be considered in all cases of unexplained hypertrophy. Its recognition is important given the advent of specific replacement enzyme therapy. PMID- 11914246 TI - Inflammation and long-term mortality after non-ST elevation acute coronary syndrome treated with a very early invasive strategy in 1042 consecutive patients. AB - BACKGROUND: This study sought to evaluate the predictive value of C-reactive protein (CRP) on long-term mortality in non-ST-elevation acute coronary syndromes (NSTACS) that were treated with a very early aggressive revascularization strategy. METHODS AND RESULTS: We conducted a prospective cohort study in 1042 consecutive patients with NSTACS who were undergoing coronary angiography and subsequent coronary stenting of the culprit lesion as the primary revascularization strategy within 24 hours. Levels of CRP were determined on admission. The patients were followed for a mean of 20 months. In-hospital mortality was significantly higher in patients with a CRP>10 mg/L (3.7% versus 1.2% with CRP<3 mg/L and versus 0.8% with CRP of 3 to 10 mg/L; relative risk for CRP>10 mg/L compared with CRP< or =10 mg/L was 4.2, 95% confidence interval [CI] was 1.6 to 11.0; P=0.004). The increase in mortality in patients with CRP>10 mg/L persisted during follow-up. Long-term mortality was 3.4% with CRP<3 mg/L, 4.4% with CRP between 3 and 10 mg/L, and 12.7% with CRP>10 mg/L (relative risk for CRP>10 mg/L compared with CRP< or =10 mg/L, 0.8; 95% CI, 2.3 to 6.2; P<0.001). In addition, Kaplan-Meier survival analysis demonstrated a significantly reduced survival at 4 years in patients with a CRP>10 mg/L (78% versus 88% for a CRP of 3 to 10 mg/L and versus 92% for CRP<3 mg/L; P<0.001 by log-rank). In a multivariate analysis, CRP was an independent predictor of long-term mortality. Patients with a CRP>10 mg/L had >4 times the risk of death (odds ratio, 4.1; 95% CI, 2.3 to 7.2). CONCLUSION: CRP is a strong independent predictor of short and long-term mortality after NSTACS that are treated with very early revascularization. PMID- 11914247 TI - Acute pulmonary embolectomy: a contemporary approach. AB - BACKGROUND: Acute pulmonary embolism confers a high mortality rate despite advances in diagnosis and therapy. Thrombolysis is often effective but has a high frequency of major bleeding complications, especially intracranial hemorrhage. Therefore, we liberalized our criteria for acute pulmonary embolectomy and considered operating on patients with anatomically extensive pulmonary embolism and concomitant moderate to severe right ventricular dysfunction despite preserved systemic arterial pressure. METHODS AND RESULTS: We report 29 (17 men and 12 women) consecutive patients who underwent embolectomy from October 1999 through October 2001. Twenty-six patients (89%) survived surgery and were alive more than 1 month postoperatively. Median follow-up is 10 months. CONCLUSION: The high survival rate of 89% can be attributed to improved surgical technique, rapid diagnosis and triage, and careful patient selection. We hope that other tertiary centers will evaluate pulmonary embolism patients with an algorithm that includes surgical embolectomy as one of several therapeutic options. Our contemporary approach to pulmonary embolectomy no longer confines this operation to a treatment of last resort reserved for clinically desperate circumstances. PMID- 11914248 TI - High-dose intravascular beta-radiation after de novo stent implantation induces coronary artery spasm. AB - BACKGROUND: Intracoronary brachytherapy is effective in preventing restenosis after coronary interventions. However, in vitro and animal studies have shown that irradiation produces immediate and sustained endothelial dysfunction. This study assesses the clinical relevance of impaired vasomotoric function induced by brachytherapy. METHODS AND RESULTS: We analyzed the occurrence of postradiation coronary artery spasms in 1 animal study and 2 clinical trials investigating the effects of high-dose intracoronary beta-radiation after de novo coronary artery stenting. Irradiated segments (IRSs) proximal and distal to the stent were studied by quantitative coronary angiography after stenting, after radiation, and at the end of the procedure. There was an 67% overall incidence of coronary artery spasm in the IRSs immediately after beta-radiation compared with 9% after sham treatment (P<0.001). Whereas in most cases this phenomenon was only minor or moderate, in 12 cases, 4 (22%) animals and 8 (28%) patients, severe coronary spasm (>90% diameter stenosis) with significant ECG-changes or hemodynamic instability was observed. Relief of spasms was protracted (mean time until complete relief of spasm 423+/-122 seconds) and required repetitive intracoronary administration of nitroglycerin (mean dose: 1.2+/-0.6 mg). CONCLUSIONS: Vasoconstriction is a frequent reaction of coronary arteries after high-dose intracoronary beta-radiation, necessitating repetitive administration of vasodilators. PMID- 11914249 TI - Coronary heart disease in patients with low LDL-cholesterol: benefit of pravastatin in diabetics and enhanced role for HDL-cholesterol and triglycerides as risk factors. AB - BACKGROUND: In two large secondary prevention trials of pravastatin, risk reduction was not significant in participants who had low baseline LDL-C concentrations (that is, <125 mg/dL). We conducted exploratory analyses of participant characteristics, lipid risk factors, and risk reduction in this group. METHODS AND RESULTS: Among 13 173 participants with coronary heart disease (CHD), 2607 had baseline LDL-C <125 mg/dL. Those with LDL-C <125 compared with > or =125 mg/dL were more likely to be diabetic (15% versus 9%), hypertensive (46 versus 41%), and male (89% versus 83%); they had higher triglycerides (169 versus 154 mg/dL), lower HDL-C (36.5 versus 38 mg/dL), and similar body mass index (27 kg/m2); and pravastatin lowered their LDL-C by 36 mg/dL (32%) versus 45 mg/dL (29%). During 5.8-year (mean) follow-up, HDL-C and triglycerides were both significantly stronger predictors of recurrent CHD events in participants with LDL-C <125 than > or =125 mg/dL. In diabetic participants with low LDL-C, pravastatin decreased CHD events from 34% to 22% (relative risk, 0.56; 95% CI, 0.37 to 0.83; P=0.004), significantly different from the effect in nondiabetic participants with low LDL-C (P interaction, 0.005) (event rate, 21%; relative risk, 1.06 [95% CI, 0.89 to 1.27]). There were trends toward risk reduction in smokers and in those with low HDL-C, <40 mg/dL. CONCLUSIONS: Among patients with CHD who have low LDL-C, diabetics have much higher subsequent CHD event rates than do nondiabetics. Pravastatin reduced the event rate in diabetics to that of nondiabetic participants. The results also suggest enhanced therapeutic potential for improving HDL-C and triglycerides in patients with CHD who have low LDL-C concentrations. PMID- 11914251 TI - Effect of medroxyprogesterone acetate on vascular inflammatory markers in postmenopausal women receiving estrogen. AB - BACKGROUND: Estrogen increases C-reactive protein (CRP) in postmenopausal women. Estrogen also decreases cell adhesion molecules, whereas elevated CRP stimulates the expression of cell adhesion molecules. Because androgens have antiinflammatory effects, androgenic progestins such as medroxyprogesterone acetate (MPA) may inhibit proinflammatory effects of estrogen. We investigated the effects of MPA on estrogen-induced changes in acute inflammatory proteins and cell adhesion molecules in postmenopausal women. METHODS AND RESULTS: Postmenopausal women were treated daily with conjugated equine estrogen (CEE, 0.625 mg), CEE plus MPA 2.5 mg, or CEE plus MPA 5.0 mg for 3 months. CEE significantly increased CRP concentrations by 320.1+/-210.2% (P<0.05). The addition of MPA to CEE, however, inhibited the increase in CRP in a concentration dependent manner (MPA 2.5 mg, 169.8+/-66.9%, P<0.05; MPA 5 mg, 55.0+/-30.4%, not significant). Similarly, CEE increased amyloid A protein concentrations, whereas MPA reversed this effect. Interleukin-6 concentration did not change significantly in any treatment group. CEE alone significantly decreased the concentration of E-selectin, but the concentrations of intercellular adhesion molecule and vascular cellular adhesion molecule did not change significantly. The addition of MPA tended to decrease the levels of cell adhesion molecules, and use of 5.0 mg MPA showed significant decreases in all cell-adhesion molecule concentrations. CONCLUSIONS: Concurrent MPA administration may attenuate estrogen's proinflammatory effect. Because MPA in combination with CEE decreased cell adhesion molecule concentrations, the anti-inflammatory effect of MPA may actually be responsible for the favorable effect of estrogen-progestogen combinations on cell adhesion molecules in postmenopausal women. PMID- 11914250 TI - Superoxide production and expression of nox family proteins in human atherosclerosis. AB - BACKGROUND: NAD(P)H oxidases are important sources of superoxide in the vasculature, the activity of which is associated with risk factors for human atherosclerosis. This study was designed to investigate the localization of superoxide production and the expression of the Nox family of NAD(P)H oxidase proteins (gp91phox, Nox1, and Nox4) in nonatherosclerotic and atherosclerotic human coronary arteries. METHODS AND RESULTS: In coronary artery segments from explanted human hearts, we examined intracellular superoxide production with dihydroethidium. In nonatherosclerotic coronary arteries, superoxide was present homogenously throughout the intima, media, and adventitia. In atherosclerotic arteries, there was an additional intense area of superoxide in the plaque shoulder, which is rich in macrophages and alpha-actin-positive cells. p22phox colocalized with gp91phox mainly in macrophages, whereas Nox4 was found only in nonphagocytic vascular cells. Expression of gp91phox and p22phox mRNA was associated with the severity of atherosclerosis. gp91phox correlated with the plaque macrophage content, whereas Nox4 correlated with the content of alpha actin-positive cells. Nox1 expression was low both in human coronary arteries and isolated vascular cells. CONCLUSIONS: Several Nox proteins, including gp91phox and Nox4, may contribute to increased intracellular oxidative stress in human coronary atherosclerosis in a cell-specific manner and thus may be involved in the genesis and progression of human coronary atherosclerotic disease. PMID- 11914253 TI - Withdrawal of statins increases event rates in patients with acute coronary syndromes. AB - BACKGROUND: HMG-CoA Reductase Inhibitors (statins) reduce cardiac event rates in patients with stable coronary heart disease. Withdrawal of chronic statin treatment during acute coronary syndromes may impair vascular function independent of lipid-lowering effects and thus increase cardiac event rate. METHODS AND RESULTS: We investigated the effects of statins on the cardiac event rate in 1616 patients of the Platelet Receptor Inhibition in Ischemic Syndrome Management (PRISM) study who had coronary artery disease and chest pain in the previous 24 hours. We recorded death and nonfatal myocardial infarction during the 30-day follow-up. Baseline clinical characteristics did not differ among 1249 patients without statin therapy, 379 patients with continued statin therapy, and 86 patients with discontinued statin therapy after hospitalization. Statin therapy was associated with a reduced event rate at 30-day follow-up compared with patients without statins (adjusted hazard ratio, 0.49 [95% CI, 0.21 to 0.86]; P=0.004). If the statin therapy was withdrawn after admission, cardiac risk increased compared with patients who continued to receive statins (2.93 [95% CI, 1.64 to 6.27]; P=0.005) and tended to be higher compared with patients who never received statins (1.69 [95% CI, 0.92 to 3.56]; P=0.15). This was related to an increased event rate during the first week after onset of symptoms and was independent of cholesterol levels. In a multivariate model, troponin T elevation (P=0.005), ST changes (P=0.02), and continuation of statin therapy (P=0.008) were the only independent predictors of patient outcome. CONCLUSIONS: Statin pretreatment in patients with acute coronary syndromes is associated with improved clinical outcome. However, discontinuation of statins after onset of symptoms completely abrogates this beneficial effect. PMID- 11914252 TI - Peroxisome proliferator-activated receptor alpha gene variants influence progression of coronary atherosclerosis and risk of coronary artery disease. AB - BACKGROUND: Peroxisome proliferator-activated receptor alpha (PPARalpha) regulates the expression of genes involved in lipid metabolism and inflammation, making it a candidate gene for atherosclerosis and ischemic heart disease (IHD). METHODS AND RESULTS: We investigated the association between the leucine 162 to valine (L162V) polymorphism and a G to C transversion in intron 7 of the PPARalpha gene and progression of atherosclerosis in the Lopid Coronary Angiography Trial (LOCAT), a trial examining the effect of gemfibrozil treatment on progression of atherosclerosis after bypass surgery and on risk of IHD in the second Northwick Park Heart Study (NPHS2), a prospective study of healthy middle aged men in the United Kingdom. There was no association with plasma lipid concentrations in either study. Both polymorphisms influenced progression of atherosclerosis and risk of IHD. V162 allele carriers had less progression of diffuse atherosclerosis than did L162 allele homozygotes with a similar trend for focal atherosclerosis. Intron 7 C allele carriers had greater progression of atherosclerosis than did G allele homozygotes. The V162 allele attenuated the proatherosclerotic effect of the intron 7 C allele. Homozygotes for the intron 7 C allele had increased risk of IHD, an effect modulated by the L162V polymorphism CONCLUSIONS: The PPARalpha gene affects progression of atherosclerosis and risk of IHD. Absence of association with plasma lipid concentrations suggests that PPARalpha affects atherosclerotic progression directly in the vessel wall. PMID- 11914254 TI - Primary prevention of sudden cardiac death in idiopathic dilated cardiomyopathy: the Cardiomyopathy Trial (CAT). AB - BACKGROUND: Patients with idiopathic dilated cardiomyopathy (DCM) and impaired left ventricular ejection fraction have an increased risk of dying suddenly. METHODS AND RESULTS: Patients with recent onset of DCM (< or =9 months) and an ejection fraction < or =30% were randomly assigned to the implantation of an implantable cardioverter-defibrillator (ICD) or control. The primary end point of the trial was all-cause mortality at 1 year of follow-up. The trial was terminated after the inclusion of 104 patients because the all-cause mortality rate at 1 year did not reach the expected 30% in the control group. In August 2000, the vital status of all patients was updated by contacting patients, relatives, or local registration offices. One hundred four patients were enrolled in the trial: Fifty were assigned to ICD therapy and 54 to the control group. Mean follow-up was 22.8+/-4.3 months, on the basis of investigators' follow-up. After 1 year, 6 patients were dead (4 in the ICD group and 2 in the control group). No sudden death occurred during the first and second years of follow-up. In August 2000, after a mean follow-up of 5.5+/-2.2 years, 30 deaths had occurred (13 in the ICD group and 17 in the control group). Cumulative survival was not significantly different between the two groups (93% and 80% in the control group versus 92% and 86% in the ICD group after 2 and 4 years, respectively). CONCLUSIONS: This trial did not provide evidence in favor of prophylactic ICD implantation in patients with DCM of recent onset and impaired left ventricular ejection fraction. PMID- 11914255 TI - Baroreflex buffering and susceptibility to vasoactive drugs. AB - BACKGROUND: The overall effect of vasoactive drugs on blood pressure is determined by a combination of the direct effect on vascular tone and an indirect baroreflex-mediated effect, a baroreflex buffering of blood pressure. Differences in baroreflex function affect the responsiveness to vasoactive medications, particularly baroreflex buffering of blood pressure; however, the magnitude is not known. METHODS AND RESULTS: We characterized baroreflex function and responses to vasoactive drugs in patients with idiopathic orthostatic intolerance, patients with essential hypertension, patients with monogenic hypertension and brachydactyly, patients with multiple system atrophy, and control subjects. We used phenylephrine sensitivity during ganglionic blockade as a measure of baroreflex buffering. Phenylephrine (25 microg) increased systolic blood pressure 6+/-1.6 mm Hg in control subjects, 6+/-1.1 mm Hg in orthostatic intolerance patients, 18+/-3.9 mm Hg in patients with essential hypertension, 31+/-3.4 mm Hg in patients with monogenic hypertension, and 25+/-3.4 mm Hg in patients with multiple system atrophy. Similar differences in sensitivities between groups were observed with nitroprusside. The sensitivity to vasoactive drugs was highly correlated with baroreflex buffering function and to a lesser degree with baroreflex control of heart rate. In control subjects, sensitivities to nitroprusside and phenylephrine infusions were correlated with baroreflex heart rate control and sympathetic nerve traffic. CONCLUSIONS: Our findings are consistent with an important effect of baroreflex blood pressure buffering on the sensitivity to vasoactive drugs. They suggest that even moderate changes in baroreflex function may have a substantial effect on the sensitivity to vasoactive medications. PMID- 11914256 TI - Which patients benefit from percutaneous mitral balloon valvuloplasty? Prevalvuloplasty and postvalvuloplasty variables that predict long-term outcome. AB - BACKGROUND: Percutaneous mitral balloon valvuloplasty (PMV) results in good immediate results, particularly in patients with echocardiographic scores (Echo Sc) < or =8. However, which variables relate to long-term outcome is unclear. METHODS AND RESULTS: We report the immediate and long-term clinical follow-up (mean, 4.2+/-3.7 years; range, 0.5 to 15) of 879 patients who underwent 939 PMV procedures. Patients were divided into 2 groups, Echo-Sc < or =8 (n=601) and Echo Sc >8 (n=278). PMV resulted in an increase in mitral valve area from 1.0+/-0.3 to 2.0+/-0.6 cm2 in patients with Echo-Sc < or =8 and from 0.8+/-0.3 to 1.6+/-0.6 cm2 in patients with Echo-Sc >8 (P<0.0001). Although adverse events (death, mitral valve surgery, and redo PMV) were low within the first 5 years of follow up, a progressive number of events occurred beyond this period. Nevertheless, survival (82% versus 57%) and event-free survival (38% versus 22%) at 12-year follow-up was greater in patients with Echo-Sc < or =8 (P<0.0001). Cox regression analysis identified post-PMV mitral regurgitation > or =3+, Echo-Sc >8, age, prior surgical commissurotomy, NYHA functional class IV, pre-PMV mitral regurgitation > or =2+, and higher post-PMV pulmonary artery pressure as independent predictors of combined events at long-term follow-up. CONCLUSIONS: The immediate and long-term outcome of patients undergoing PMV is multifactorial. The use of the Echo-Sc in conjunction with other clinical and morphological predictors of PMV outcome allows identification of patients who will obtain the best outcome from PMV. PMID- 11914257 TI - New Bayesian discriminator for detection of atrial tachyarrhythmias. AB - BACKGROUND: Accurate, rapid detection of atrial tachyarrhythmias has important implications in the use of implantable devices for treatment of cardiac arrhythmia. Currently available detection algorithms for atrial tachyarrhythmias, which use the single-index method, have limited sensitivity and specificity. METHODS AND RESULTS: In this study, we evaluated the performance of a new Bayesian discriminator algorithm in the detection of atrial fibrillation (AF), atrial flutter (AFL), and sinus rhythm (SR). Bipolar recording of 364 rhythms (AF=156, AFL=88, SR=120) at the high right atrium were collected from 20 patients who underwent electrophysiological procedures. After initial signal processing, a column vector of 5 features for each rhythm were established, based on the regularity, rate, energy distribution, percent time of quiet interval, and baseline reaching of the rectified autocorrelation coefficient functions. Rhythm identification was obtained by use of Bayes decision rule and assumption of Gaussian distribution. For the new Bayesian discriminator, the overall sensitivity for detection of SR, AF, and AFL was 97%, 97%, and 94%, respectively; and the overall specificity for detection of SR, AF, and AFL was 98%, 98%, and 99%, respectively. The overall accuracy of detection of SR, AF, and AFL was 98%, 97% and 98%, respectively. Furthermore, sensitivity, specificity, and accuracy of this algorithm were not affected by a range of white Gaussian noises with different intensities. CONCLUSIONS: This new Bayesian discriminator algorithm, based on Bayes decision of multiple features of atrial electrograms, allows rapid on-line and accurate (98%) detection of AF with robust anti-noise performance. PMID- 11914258 TI - Switching off embolization from symptomatic carotid plaque using S nitrosoglutathione. AB - BACKGROUND: Current antiplatelet regimens fail to prevent the majority of recurrent strokes. Asymptomatic circulating emboli can be detected by transcranial Doppler ultrasound, are frequent in patients with symptomatic carotid stenosis, and predict recurrent stroke risk. S-nitrosoglutathione (GSNO) is a nitric oxide donor that appears to have relative platelet specificity. We evaluated its effectiveness in reducing embolization in patients with symptomatic carotid stenosis who already were taking aspirin. METHODS AND RESULTS: Twenty patients with > or =50% internal carotid artery stenosis and with > or =3 embolic signals detected during a half-hour screening recording were recruited. All had taken aspirin for at least 7 days. They were randomly assigned in a double-blind fashion to either GSNO (4.4 mmol/kg per minute) or saline placebo for 90 minutes. Transcranial Doppler recordings were made from the ipsilateral middle cerebral artery for 1 hour before treatment and at 0 to 3, 6, and 24 hours after treatment. Before treatment, the mean (range) of embolic signals per hour was 6.9 (3 to 13) in the GSNO group and 7.3 (4 to 12) in the placebo group (P=0.68). GSNO resulted in a rapid reduction in the frequency of embolic signals of 84% at 0 to 3 hours, 95% at 6 hours, and 100% at 24 hours (P<0.0001, P=0.003, and P<0.0001 versus placebo, respectively). CONCLUSIONS: Continued embolization is common in patients with carotid stenosis despite aspirin therapy. GSNO was highly effective in rapidly reducing the frequency of embolic signals in this patient group. Despite its short administration time and its short half-life, it resulted in therapeutic effects lasting 24 hours. PMID- 11914259 TI - Vulnerable atherosclerotic plaque morphology in apolipoprotein E-deficient mice unable to make ascorbic Acid. AB - BACKGROUND: Oxidative stress is thought to play an important role in atherogenesis, suggesting that antioxidants could prevent coronary artery disease. However, the efficacy of vitamin C in reducing atherosclerosis is debatable in humans and has not been tested rigorously in animals. METHODS AND RESULTS: Gulo(-/-)Apoe(-/-) mice were used to test a hypothesis that chronic vitamin C deficiency enhances the initiation and development of atherosclerosis. These mice are dependent on dietary vitamin C because of the lack of L gulonolactone-gamma-oxidase and are prone to develop atherosclerosis because of lacking apolipoprotein E. Beginning at 6 weeks of age, the Gulo(-/-)Apoe(-/-) mice were fed regular chow or Western-type diets containing high fat and supplemented with either 0.033 g or 3.3 g/L of vitamin C in their drinking water. This regimen produced mice with chronically low vitamin C (average 1.5 microg/mL in plasma) or high vitamin C (average 10 to 30 microg/mL in plasma). Morphometric analysis showed that within each sex, age, and diet group, the sizes of the atherosclerotic plaques were not different between low vitamin C mice and high vitamin C mice. However, advanced plaques in the low vitamin C mice had significantly reduced amounts of Sirius red-staining collagen (36.4+/-2.2% versus 54.8+/-2.3%, P<0.0001), larger necrotic cores within the plaques, and reduced fibroproliferation and neovascularization in the aortic adventitia. CONCLUSIONS: Chronic vitamin C deficiency does not influence the initiation or progression of atherosclerotic plaques but severely compromises collagen deposition and induces a type of plaque morphology that is potentially vulnerable to rupture. PMID- 11914260 TI - Impairment of collateral formation in lipoprotein(a) transgenic mice: therapeutic angiogenesis induced by human hepatocyte growth factor gene. AB - BACKGROUND: Although lipoprotein(a) (Lp[a]) is a risk factor for atherosclerosis, no study has documented the effects of Lp(a) on angiogenesis. In this study, we examined collateral formation in peripheral arterial disease (PAD) model in Lp(a) transgenic mice. In addition, we examined the feasibility of gene therapy by using an angiogenic growth factor, hepatocyte growth factor (HGF), to treat PAD in the presence of high Lp(a). METHODS AND RESULTS: In Lp(a) transgenic mice, the degree of natural recovery of blood flow after operation was significantly lower than that in nontransgenic mice. Of importance, there was a significant negative correlation between serum Lp(a) concentration and the degree of natural recovery of blood flow (P<0.05). In addition, Lp(a) significantly stimulated the growth of vascular smooth muscle, accompanied by the phosphorylation of ERK. These data demonstrated the association of impairment of collateral formation with serum Lp(a) concentration. Thus, we examined the feasibility of therapeutic angiogenesis by using HGF, with the goal of progression to human gene therapy. Intramuscular injection of HGF plasmid resulted in a significant increase in blood flow even in Lp(a) transgenic mice, accompanied by the detection of human HGF protein. A significant increase in capillary density also was detected in Lp(a) transgenic mice transfected with human HGF compared with control (P<0.01). CONCLUSIONS: Overall, a high serum Lp(a) concentration impaired collateral formation. Although the delay of angiogenesis in high serum Lp(a) might diminish angiogenesis, intramuscular injection of HGF plasmid induced therapeutic angiogenesis in the Lp(a) transgenic ischemic hindlimb mouse model as potential therapy for PAD. PMID- 11914262 TI - New concepts in diastolic dysfunction and diastolic heart failure: Part II: causal mechanisms and treatment. PMID- 11914261 TI - Nitric oxide mediates the antiapoptotic effect of insulin in myocardial ischemia reperfusion: the roles of PI3-kinase, Akt, and endothelial nitric oxide synthase phosphorylation. AB - BACKGROUND: Recent evidence from cultured endothelial cell studies suggests that phosphorylation of endothelial nitric oxide synthase (eNOS) through the PI3 kinase-Akt pathway increases NO production. This study was designed to elucidate the signaling pathway involved in the antiapoptotic effect of insulin in vivo and to test the hypothesis that phosphorylation of eNOS by insulin may participate in the cardioprotective effect of insulin after myocardial ischemia and reperfusion. METHODS AND RESULTS: Male Sprague-Dawley rats were subjected to 30 minutes of myocardial ischemia and 4 hours of reperfusion. Rats were randomized to receive vehicle, insulin, insulin plus wortmannin, or insulin plus L-NAME. Treatment with insulin resulted in 2.6-fold and 4.3-fold increases in Akt and eNOS phosphorylation and a significant increase in NO production in ischemic/reperfused myocardial tissue. Phosphorylation of Akt and eNOS and increase of NO production by insulin were completely blocked by wortmannin, a PI3 kinase inhibitor. Pretreatment with L-NAME, a nonselective NOS inhibitor, had no effect on Akt and eNOS phosphorylation but significantly reduced NO production. Moreover, treatment with insulin markedly reduced myocardial apoptotic death (P<0.01 versus vehicle). Pretreatment with wortmannin abolished the antiapoptotic effect of insulin. Most importantly, pretreatment with L-NAME also significantly reduced the antiapoptotic effect of insulin (P<0.01 versus insulin). CONCLUSIONS: These results demonstrated that in vivo administration of insulin activated Akt through the PI3-kinase-dependent mechanism and reduced postischemic myocardial apoptotic death. Phosphorylation of eNOS and the concurrent increase of NO production contribute significantly to the antiapoptotic effect of insulin. PMID- 11914263 TI - Thoughts on the role of the healing professions and the events of September 11, 2001. PMID- 11914264 TI - Images in cardiovascular medicine. Septal course of the left main coronary artery originating from the right sinus of valsalva. PMID- 11914265 TI - Images in cardiovascular medicine. Phenotype of infiltrating T lymphocytes in cardiac sarcoidosis. PMID- 11914266 TI - Risks in risk stratification: what is relevant in practice? PMID- 11914267 TI - Hormone replacement therapy and cardiovascular disease: a statement for healthcare professionals from the American Heart Association. PMID- 11914268 TI - Prognostic implications of cardiac marker elevation after percutaneous coronary intervention. PMID- 11914269 TI - Abciximab suppresses the rise in levels of circulating inflammatory markers after percutaneous coronary revascularization. PMID- 11914270 TI - Late-breaking trials from the American College of Cardiology. PMID- 11914271 TI - The origin recognition complex: from simple origins to complex functions. PMID- 11914272 TI - Rescue of neural tube defects in Pax-3-deficient embryos by p53 loss of function: implications for Pax-3- dependent development and tumorigenesis. AB - Pax-3 is a transcription factor that is expressed in the neural tube, neural crest, and dermomyotome. We previously showed that apoptosis is associated with neural tube defects (NTDs) in Pax-3-deficient Splotch (Sp/Sp) embryos. Here we show that p53 deficiency, caused by germ-line mutation or by pifithrin-alpha, an inhibitor of p53-dependent apoptosis, rescues not only apoptosis, but also NTDs, in Sp/Sp embryos. Pax-3 deficiency had no effect on p53 mRNA, but increased p53 protein levels. These results suggest that Pax-3 regulates neural tube closure by inhibiting p53-dependent apoptosis, rather than by inducing neural tube-specific gene expression. PMID- 11914273 TI - A senescence rescue screen identifies BCL6 as an inhibitor of anti-proliferative p19(ARF)-p53 signaling. AB - Senescence limits the proliferative capacity of primary cells in culture. We describe here a genetic screen to identify genes that allow bypass of this checkpoint. Using retroviral cDNA expression libraries, we identify BCL6 as a potent inhibitor of senescence. BCL6 is frequently activated in non-Hodgkin's lymphoma, but its mechanism of action has remained unclear. BCL6 efficiently immortalizes primary mouse embryonic fibroblasts and cooperates with RAS in oncogenic transformation. BCL6 overrides the senescence response downstream of p53 through a process that requires induction of cyclin D1 expression, as cyclin D1 knockout fibroblasts are specifically resistant to BCL6 immortalization. We show that BCL6 expression also dramatically extends the replicative lifespan of primary human B cells in culture and induces cyclin D1 expression, indicating that BCL6 has a similar activity in lymphoid cells. Our results suggest that BCL6 contributes to oncogenesis by rendering cells unresponsive to antiproliferative signals from the p19(ARF)-p53 pathway. PMID- 11914274 TI - Specific targeting and constitutive association of histone deacetylase complexes during transcriptional repression. AB - Specific recruitment of corepressor complexes containing histone deacetylases (HDAC) by transcription factors is believed to play an essential role in transcriptional repression. Recent studies indicate that repression by unliganded nuclear hormone receptors and by the Mad family of repressors requires distinct HDAC-containing corepressor complexes. In this work, we show that unliganded TR specifically recruits only the closely related N-CoR and SMRT-HDAC3 complexes, whereas the Mad1 recruits only the Sin3-HDAC1/2 complex. Significantly, both the Sin3 and Mi-2/NURD complexes also exhibit constitutive association with chromatin and contribute to chromatin deacetylation in a nontargeted fashion. These results suggest that HDAC complexes can contribute to gene repression by two distinct mechanisms as follows: (1) specific targeting by repressors and (2) constitutive association with chromatin. PMID- 11914275 TI - Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. AB - The epithelial compartment of the human breast comprises two distinct lineages: the luminal epithelial and the myoepithelial lineage. We have shown previously that a subset of the luminal epithelial cells could convert to myoepithelial cells in culture signifying the possible existence of a progenitor cell. We therefore set out to identify and isolate the putative precursor in the luminal epithelial compartment. Using cell surface markers and immunomagnetic sorting, we isolated two luminal epithelial cell populations from primary cultures of reduction mammoplasties. The major population coexpresses sialomucin (MUC(+)) and epithelial-specific antigen (ESA(+)) whereas the minor population has a suprabasal position and expresses epithelial specific antigen but no sialomucin (MUC(-)/ESA(+)). Two cell lines were further established by transduction of the E6/E7 genes from human papilloma virus type 16. Both cell lines maintained a luminal epithelial phenotype as evidenced by expression of the tight junction proteins, claudin-1 and occludin, and by generation of a high transepithelial electrical resistance on semipermeable filters. Whereas in clonal cultures, the MUC(+)/ESA(+) epithelial cell line was luminal epithelial restricted in its differentiation repertoire, the suprabasal-derived MUC(-)/ESA(+) epithelial cell line was able to generate itself as well as MUC(+)/ESA(+) epithelial cells and Thy-1(+)/alpha-smooth muscle actin(+) (ASMA(+)) myoepithelial cells. The MUC( )/ESA(+) epithelial cell line further differed from the MUC(+)/ESA(+) epithelial cell line by the expression of keratin K19, a feature of a subpopulation of epithelial cells in terminal duct lobular units in vivo. Within a reconstituted basement membrane, the MUC(+)/ESA(+) epithelial cell line formed acinus-like spheres. In contrast, the MUC(-)/ESA(+) epithelial cell line formed elaborate branching structures resembling uncultured terminal duct lobular units both by morphology and marker expression. Similar structures were obtained by inoculating the extracellular matrix-embedded cells subcutaneously in nude mice. Thus, MUC( )/ESA(+) epithelial cells within the luminal epithelial lineage may function as precursor cells of terminal duct lobular units in the human breast. PMID- 11914276 TI - Subcellular localization of the yeast proteome. AB - Protein localization data are a valuable information resource helpful in elucidating eukaryotic protein function. Here, we report the first proteome-scale analysis of protein localization within any eukaryote. Using directed topoisomerase I-mediated cloning strategies and genome-wide transposon mutagenesis, we have epitope-tagged 60% of the Saccharomyces cerevisiae proteome. By high-throughput immunolocalization of tagged gene products, we have determined the subcellular localization of 2744 yeast proteins. Extrapolating these data through a computational algorithm employing Bayesian formalism, we define the yeast localizome (the subcellular distribution of all 6100 yeast proteins). We estimate the yeast proteome to encompass approximately 5100 soluble proteins and >1000 transmembrane proteins. Our results indicate that 47% of yeast proteins are cytoplasmic, 13% mitochondrial, 13% exocytic (including proteins of the endoplasmic reticulum and secretory vesicles), and 27% nuclear/nucleolar. A subset of nuclear proteins was further analyzed by immunolocalization using surface-spread preparations of meiotic chromosomes. Of these proteins, 38% were found associated with chromosomal DNA. As determined from phenotypic analyses of nuclear proteins, 34% are essential for spore viability--a percentage nearly twice as great as that observed for the proteome as a whole. In total, this study presents experimentally derived localization data for 955 proteins of previously unknown function: nearly half of all functionally uncharacterized proteins in yeast. To facilitate access to these data, we provide a searchable database featuring 2900 fluorescent micrographs at http://ygac.med.yale.edu. PMID- 11914277 TI - miRNPs: a novel class of ribonucleoproteins containing numerous microRNAs. AB - Gemin3 is a DEAD-box RNA helicase that binds to the Survival of Motor Neurons (SMN) protein and is a component of the SMN complex, which also comprises SMN, Gemin2, Gemin4, Gemin5, and Gemin6. Reduction in SMN protein results in Spinal muscular atrophy (SMA), a common neurodegenerative disease. The SMN complex has critical functions in the assembly/restructuring of diverse ribonucleoprotein (RNP) complexes. Here we report that Gemin3 and Gemin4 are also in a separate complex that contains eIF2C2, a member of the Argonaute protein family. This novel complex is a large approximately 15S RNP that contains numerous microRNAs (miRNAs). We describe 40 miRNAs, a few of which are identical to recently described human miRNAs, a class of small endogenous RNAs. The genomic sequences predict that miRNAs are likely to be derived from larger precursors that have the capacity to form stem-loop structures. PMID- 11914278 TI - C. elegans condensin promotes mitotic chromosome architecture, centromere organization, and sister chromatid segregation during mitosis and meiosis. AB - Chromosome segregation and X-chromosome gene regulation in Caenorhabditis elegans share the component MIX-1, a mitotic protein that also represses X-linked genes during dosage compensation. MIX-1 achieves its dual roles through interactions with different protein partners. To repress gene expression, MIX-1 acts in an X chromosome complex that resembles the mitotic condensin complex yet lacks chromosome segregation function. Here we show that MIX-1 interacts with a mitotic condensin subunit, SMC-4, to achieve chromosome segregation. The SMC-4/MIX-1 complex positively supercoils DNA in vitro and is required for mitotic chromosome structure and segregation in vivo. Thus, C. elegans has two condensin complexes, one conserved for mitosis and another specialized for gene regulation. SMC-4 and MIX-1 colocalize with centromere proteins on condensed mitotic chromosomes and are required for the restricted orientation of centromeres toward spindle poles. This cell cycle-dependent localization requires AIR-2/AuroraB kinase. Depletion of SMC-4/MIX-1 causes aberrant mitotic chromosome structure and segregation, but not dramatic decondensation at metaphase. Moreover, SMC-4/MIX-1 depletion disrupts sister chromatid segregation during meiosis II but not homologous chromosome segregation during meiosis I, although both processes require chromosome condensation. These results imply that condensin is not simply required for compaction, but plays a more complex role in chromosome architecture that is essential for mitotic and meiotic sister chromatid segregation. PMID- 11914281 TI - Where now for meta-analysis? PMID- 11914279 TI - Dynamics of global histone acetylation and deacetylation in vivo: rapid restoration of normal histone acetylation status upon removal of activators and repressors. AB - DNA-binding activators and repressors recruit histone acetylases and deacetylases to promoters, thereby generating localized domains of modified histones that influence transcriptional activity. At the end of a transcriptional response, alterations in histone acetylation status are reversed, but the dynamics of this process are poorly understood. Here, we recruit histone deacetylases and acetylases to a well-defined yeast promoter in a regulated manner. Following dissociation of the recruiting protein from the promoter, targeted deacetylation and acetylation are reversed with rapid, yet distinct, kinetics. Reversal of targeted deacetylation occurs within 5-8 min, whereas reversal of targeted acetylation is more rapid, taking 1.5 min. These findings imply that untargeted, globally acting enzymes generate a highly dynamic equilibrium of histone acetylation and deacetylation reactions across chromatin. Targeted acetylases and deacetylases can locally perturb this equilibrium, yet once they are removed, the global activities mediate a rapid return to the steady-state level of histone acetylation. Our results also indicate that TBP occupancy depends on the presence of the activator, not histone acetylation status. PMID- 11914282 TI - Systematic reviews in epidemiology: why are we so far behind? PMID- 11914280 TI - FLOOZY of petunia is a flavin mono-oxygenase-like protein required for the specification of leaf and flower architecture. AB - The mechanisms that determine the relative positions of floral organs, and thereby their numbers, is a poorly understood aspect of flower development. We isolated a petunia mutant, floozy (fzy), in which the formation of floral organ primordia in the outermost three floral whorls and one of the two bracts at the base of the flower is blocked at an early stage. In addition, fzy mutants fail to generate secondary veins in leaves and bracts and display a decreased apical dominance in the inflorescence. FZY encodes an enzyme with homology to flavin mono-oxygenases and appears to be the ortholog of YUCCA genes of Arabidopsis. FZY is expressed in young leafs and bracts and in developing flowers. In young floral meristems FZY is expressed in the center of the meristem dome and, later, expression becomes localized on the flanks of the initiating petal and stamen primordia and at several sites in maturing anthers and carpels. These findings indicate that FZY is involved in synthesizing a signaling compound that is required for floral organ initiation and specification of the vascularization pattern in leaves. Although fzy mutants contain normal auxin levels, ectopic expression of FZY results in excessive auxin accumulation, suggesting that the signaling compound is auxin. PMID- 11914283 TI - Civilization and peptic ulcer. 1962. PMID- 11914284 TI - Commentary: civilization and peptic ulcer 40 years on. PMID- 11914285 TI - Commentary: Helicobacter as the 'environmental factor' in Susser and Stein's cohort theory of peptic ulcer disease. PMID- 11914286 TI - Commentary: the unresolved mystery of birth-cohort phenomena in gastroenterology. PMID- 11914287 TI - Commentary: peptic ulcer, Susser and Stein and the cohort phenomenon. PMID- 11914288 TI - Commentary: peptic ulcer and its discontents. PMID- 11914289 TI - Zena Stein, Mervyn Susser and epidemiology: observation, causation and action. PMID- 11914290 TI - The evolving HIV epidemic in South Africa. PMID- 11914291 TI - From Susser's causal paradigms to social justice in Australia? PMID- 11914292 TI - Health rights for women in the age of AIDS. PMID- 11914293 TI - Two lives, three legs, one journey: a retrospective appreciation of Zena Stein and Mervyn Susser. PMID- 11914294 TI - Genomic sequencing in the service of human rights. PMID- 11914295 TI - Homocyst(e)ine and cardiovascular disease: a systematic review of the evidence with special emphasis on case-control studies and nested case-control studies. AB - BACKGROUND: Elevated concentrations of homocyst(e)ine are thought to increase the risk of vascular diseases including coronary heart disease and cerebrovascular disease. METHODS: We searched MEDLINE (1966-1999), EMBASE (1974-1999), SciSearch (1974- 1999), and Dissertation Abstracts (1999) for articles and theses about homocyst(e)ine concentration and coronary heart disease and cerebrovascular disease. RESULTS: We included 57 publications (3 cohort studies, 12 nested case control studies, 42 case-control studies) that reported results on 5518 people with coronary heart disease (11,068 control subjects) and 1817 people with cerebrovascular disease (4787 control subjects) in our analysis. For coronary heart disease, the summary odds ratios (OR) for a 5-micromol/l increase in homocyst(e)ine concentration were 1.06 (95% CI : 0.99-1.13) for 2 publications of cohort studies, 1.23 (95% CI : 1.07-1.41) for 10 publications of nested case control studies, and 1.70 (95% CI : 1.50-1.93) for 26 publications of case control studies. For cerebrovascular disease, the summary OR for a 5-micromol/l increase in homocyst(e)ine concentration were 1.10 (95% CI : 0.94-1.28) for 2 publications of cohort studies, 1.58 (95% CI : 1.35-1.85) for 5 publications of nested case-control studies, and 2.16 (95% CI : 1.65-2.82) for 17 publications of case-control studies. CONCLUSIONS: Prospective studies offer weaker support than case-control studies for an association between homocyst(e)ine concentration and cardiovascular disease. Although other lines of evidence support a role for homocyst(e)ine in the pathogenesis of cardiovascular disease, more information from prospective epidemiological studies or clinical trials is needed to clarify this role. PMID- 11914296 TI - Commentary: an updated review of the published studies of homocysteine and cardiovascular disease. PMID- 11914297 TI - Can we individualize the 'number needed to treat'? An empirical study of summary effect measures in meta-analyses. AB - BACKGROUND: Meta-analyses summarize the magnitude of treatment effect using a number of measures of association, including the odds ratio (OR), risk ratio (RR), risk difference (RD) and/or number needed to treat (NNT). In applying the results of a meta-analysis to individual patients, some textbooks of evidence based medicine advocate individualizing NNT, based on the RR and the patient's expected event rate (PEER). This approach assumes constant RR but no empirical study to date has examined the validity of this assumption. METHODS: We randomly selected a subset of meta-analyses from a recent issue of the Cochrane Library (1998, Issue 3). When a meta-analysis pooled more than three randomized controlled trials (RCT) to produce a summary measure for an outcome, we compared the OR, RR and RD of each RCT with the corresponding pooled OR, RR and RD from the meta-analysis of all the other RCT. Using the conventional P-value of 0.05, we calculated the percentage of comparisons in which there were no statistically significant differences in the estimates of OR, RR or RD, and refer to this percentage as the 'concordance rate'. RESULTS: For each effect measure, we made 1843 comparisons, extracted from 55 meta-analyses. The random effects model OR had the highest concordance rate, closely followed by the fixed effects model OR and random effects model RR. The minimum concordance rate for these indices was 82%, even when the baseline risk differed substantially. The concordance rates for RD, either fixed effects or random effects model, were substantially lower (54-65%). CONCLUSIONS: The fixed effects OR, random effects OR and random effects RR appear to be reasonably constant across different baseline risks. Given the interpretational and arithmetic ease of RR, clinicians may wish to rely on the random effects model RR and use the PEER to individualize NNT when they apply the results of a meta-analysis in their practice. PMID- 11914298 TI - Commentary: relative treatment effects are consistent across the spectrum of underlying risks...usually. PMID- 11914300 TI - Commentary: improving pooled analyses in epidemiology. PMID- 11914299 TI - Meat and dairy food consumption and breast cancer: a pooled analysis of cohort studies. AB - BACKGROUND: More than 20 studies have investigated the relation between meat and dairy food consumption and breast cancer risk with conflicting results. Our objective was to evaluate the risk of breast cancer associated with meat and dairy food consumption and to assess whether non-dietary risk factors modify the relation. METHODS: We combined the primary data from eight prospective cohort studies from North America and Western Europe with at least 200 incident breast cancer cases, assessment of usual food and nutrient intakes, and a validation study of the dietary assessment instrument. The pooled database included 351,041 women, 7379 of whom were diagnosed with invasive breast cancer during up to 15 years of follow-up. RESULTS: We found no significant association between intakes of total meat, red meat, white meat, total dairy fluids, or total dairy solids and breast cancer risk. Categorical analyses suggested a J-shaped association for egg consumption where, compared to women who did not eat eggs, breast cancer risk was slightly decreased among women who consumed < 2 eggs per week but slightly increased among women who consumed > or = 1 egg per day. CONCLUSIONS: We found no significant associations between intake of meat or dairy products and risk of breast cancer. An inconsistent relation between egg consumption and risk of breast cancer merits further investigation. PMID- 11914301 TI - Asymmetric funnel plots and publication bias in meta-analyses of diagnostic accuracy. AB - BACKGROUND: Despite the great possibility of publication bias in studies of diagnostic test research, empirical studies about publication bias have mainly focused on studies of treatment effect. METHODS: A sample of 28 meta-analyses of diagnostic accuracy was selected from the Database of Abstracts of Reviews of Effectiveness (DARE). Methods used to deal with publication and related biases in these meta-analyses were examined. Asymmetry of funnel plot of estimated test accuracy against corresponding precision for each meta-analysis was assessed by three statistical methods: rank correlation method, regression analysis, and Trim and Fill method. RESULTS: In reviews of diagnostic accuracy, there was a general lack of consideration of appropriate literature searching to minimize publication bias, and the impact of possible publication bias has not been systematically assessed. The results of the three different statistical methods consistently showed that in a large proportion of the 28 meta-analyses evaluated, the smaller studies were associated with a greater diagnostic accuracy. Exploratory analyses found that the fewer the literature databases searched, the greater the funnel plot asymmetry in meta-analyses. Funnel plot asymmetry tended to be greater in meta-analyses that included smaller number of primary studies. Our data revealed no consistent relationship between funnel plot asymmetry and language restriction in reviews. CONCLUSIONS: Further research is required to explain why smaller studies tended to report greater test accuracy in a large proportion of meta analyses of diagnostic tests. In systematic reviews of diagnostic studies, literature search should be sufficiently comprehensive and possible impact of publication bias should be assessed. PMID- 11914302 TI - Being sceptical about meta-analyses: a Bayesian perspective on magnesium trials in myocardial infarction. AB - BACKGROUND: There has been extensive discussion of the apparent conflict between meta-analyses and a mega-trial investigating the benefits of intravenous magnesium following myocardial infarction, in which the early trial results have been said to be 'too good to be true'. METHODS: We apply Bayesian methods of meta analysis to the trials available before and after the publication of the ISIS-4 results. We show how scepticism can be formally incorporated into an analysis as a Bayesian prior distribution, and how Bayesian meta-analysis models allow appropriate exploration of hypotheses that the treatment effect depends on the size of the trial or the risk in the control group. RESULTS: Adoption of a sceptical prior would have led early enthusiasm for magnesium to be suitably tempered, but only if combined with a random effects meta-analysis, rather than the fixed effect analysis that was actually conducted. CONCLUSIONS: We argue that neither a fixed effect nor a random effects analysis is appropriate when the mega trial is included. The Bayesian framework provides many possibilities for flexible exploration of clinical hypotheses, but there can be considerable sensitivity to apparently innocuous assumptions. PMID- 11914303 TI - Commentary: biostatistics, biological mechanisms and Bayes: lessons from the magnesium trials. PMID- 11914304 TI - Effects of adjusting for censoring on meta-analyses of time-to-event outcomes. AB - BACKGROUND: Systematic reviews of published time-to-event outcomes commonly rely on calculating odds ratios (OR) at fixed points in time and where actual numbers at risk are not presented. These estimates are usually based on the total numbers included in the published analysis and take no account of censoring. We have assessed the impact of adjusting for censoring on weighting, estimates and statistical heterogeneity of meta-analyses in cancer. METHODS: Meta-analyses of survival data for five meta-analyses of published trials in cancer were conducted. The OR and associated statistics were calculated based on unadjusted total numbers of participants and events. These were compared with calculations that first adjusted the numbers at risk for censoring using a simple model. RESULTS: Pooled OR were changed in 17/24 cases. On average, there was a 2.6% difference between the adjusted and unadjusted OR. Confidence intervals were frequently wider for the adjusted OR. Adjusting also reduced weighting of individual trials with immature follow-up. In 18/24 cases, adjusting reduced statistical heterogeneity and affected the associated P-values. CONCLUSIONS: Reviewers conducting meta-analyses of published time-to-event data where actual numbers at risk are not available should adjust the numbers at risk, estimated from total numbers analysed, to account for immature data and censoring. PMID- 11914305 TI - Searching the Literatura Latino Americana e do Caribe em Ciencias da Saude (LILACS) database improves systematic reviews. AB - BACKGROUND: An unbiased systematic review (SR) should analyse as many articles as possible in order to provide the best evidence available. However, many SR use only databases with high English-language content as sources for articles. Literatura Latino Americana e do Caribe em Ciencias da Saude (LILACS) indexes 670 journals from the Latin American and Caribbean health literature but is seldom used in these SR. Our objective is to evaluate if LILACS should be used as a routine source of articles for SR. METHODS: First we identified SR published in 1997 in five medical journals with a high impact factor. Then we searched LILACS for articles that could match the inclusion criteria of these SR. We also checked if the authors had already identified these articles located in LILACS. RESULTS: In all, 64 SR were identified. Two had already searched LILACS and were excluded. In 39 of 62 (63%) SR a LILACS search identified articles that matched the inclusion criteria. In 5 (8%) our search was inconclusive and in 18 (29%) no articles were found in LILACS. Therefore, in 71% (44/72) of cases, a LILACS search could have been useful to the authors. This proportion remains the same if we consider only the 37 SR that performed a meta-analysis. In only one case had the article identified in LILACS already been located elsewhere by the authors' strategy. CONCLUSION: LILACS is an under-explored and unique source of articles whose use can improve the quality of systematic reviews. This database should be used as a routine source to identify studies for systematic reviews. PMID- 11914306 TI - Direction and impact of language bias in meta-analyses of controlled trials: empirical study. AB - BACKGROUND: Excluding clinical trials reported in languages other than English from meta-analyses may introduce bias and reduce the precision of combined estimates of treatment effects. We examined the influence of trials published in languages other than English on combined estimates and conclusions of published meta-analyses. METHODS: We searched journals and the Cochrane Database of Systematic Reviews for meta-analyses of at least five trials with binary outcomes that were based on comprehensive literature searches without language restrictions. We compared estimates of treatment effects from trials published in languages other than English to those from trials published in English, and assessed the impact of restricting meta-analyses to trials published in English. RESULTS: We identified 303 meta-analyses: 159 (52.4%) employed comprehensive literature searches of which 50 included 485 English and 115 non-English language trials. Non-English language trials included fewer participants (median 88 versus 116, P = 0.006) and were more likely to produce significant results at P < 0.05 (41.7% versus 31.3%, P = 0.033). The methodological quality of non-English language trials tended to be lower than that of trials published in English. Estimates of treatment effects were on average 16% (95% CI : 3-26%) more beneficial in non-English-language trials than in English-language trials. In 29 (58.0%) meta-analyses the change in effect estimates after exclusion of non English language trials was less than 5%. In the remaining meta-analyses, 5 (10.0%) showed more benefit and 16 (32.0%) less benefit after exclusion of non English language trials. CONCLUSIONS: This retrospective analysis suggests that excluding trials published in languages other than English has generally little effect on summary treatment effect estimates. The importance of non-English language trials is, however, difficult to predict for individual systematic reviews. Comprehensive literature searches followed by a careful assessment of trial quality are required to assess the contribution of all relevant trials, independent of language of publication. PMID- 11914307 TI - Commentary: searching for trials for systematic reviews: what difference does it make? PMID- 11914308 TI - Number and size of randomized trials reported in general health care journals from 1948 to 1997. AB - BACKGROUND: Randomized trials are important for controlling selection biases, and where sufficient numbers of participants are involved, have the potential to yield reliable estimates of treatment effects. METHODS: We investigated trends in the number and size of randomized trials reported in general health care journals from 1948 to 1997. From the handsearching of 18 general health care journals we collected data on the number of reports of randomized trials in each journal per year, and the number of participants in each trial. RESULTS: A total of 5503 reports of trials were identified in 18 general health care journals. More than a third appeared in the British Medical Journal. The peak period for trial reports was the mid 1980s, with more in 1986 than any other year (242). By the mid 1990s the number per year had declined by a third. Trials with fewer than 100 participants accounted for most of the reports (69%). In spite of the overall decline in the number of trial reports, those involving 100 participants or more continued to increase throughout the period studied. CONCLUSIONS: The continued increase in the number of larger trials reported is encouraging, especially if it represents an increase in the size of trials more generally. Further research is needed to determine whether the trends over time identified here are reflective more of trends in the actual conduct of, rather than simply the reporting, of randomized trials. PMID- 11914309 TI - Sources of variation of Helicobacter pylori treatment success in adults worldwide: a meta-analysis. AB - BACKGROUND: A vast number of Helicobacter pylori treatment trials have been conducted. Regimens may vary in efficacy in different patient populations. METHODS: We identified sources of treatment effect variation from 618 treatment groups using weighted cross-classified multi-level meta-regression models. Summary effect estimates were calculated within groups that lacked identified heterogeneity. RESULTS: Overall, treatment was less successful with shorter treatment duration and dual drug (versus triple or quadruple drug) therapies. For nitroimidazole-based regimens, treatment was less successful in populations with frequent childhood H. pylori infection or metronidazole resistance and more successful in northeastern Asia. Non-nitroimidazole treatments of longer duration and those from less recent reports were most successful. Some one-week regimens- (nitroimidazole/ tetracycline/bismuth, ranitidine bismuth citrate/amoxicillin/clarithromycin, and clarithromycin/amoxicillin/proton pump inhibitor) were highly successful in northeastern Asia regardless of metronidazole resistance. The most successful regimen in populations with both a high prevalence of metrondiazole resistance and frequent infection in children (metronidazole/furazolidone/amoxicillin) eliminated fewer than 70% of infections. CONCLUSIONS: More effective treatments are needed for most populations of the world where H. pylori infection in children and drug resistance are common. Current treatment guidelines do not coincide with the best treatment regimens identified in this meta-analysis. PMID- 11914310 TI - Meta-analyses involving cross-over trials: methodological issues. AB - BACKGROUND: Meta-analysis of randomized controlled trials (RCTs) is usually based on trials where patients are randomized individually into two different, parallel, treatment groups. This paper concentrates on RCTs of a different design two-period, two-treatment cross-over trials. METHODS: The characteristics of these trials are outlined, with detailed examples of methods for analysis for both continuous and binary data. These case studies are then extended into the context of a meta-analysis. The Cochrane Library was surveyed to assess current practice for synthesis. RESULTS: Methods are described for continuous and binary data for use both when the necessary paired data are given and also when they need to be calculated or imputed, and some suggestions are provided to help people wishing to synthesize data from cross-over trials into meta-analyses. The survey suggested that about 8% of the trials in the Cochrane library were cross over trials and 18% of the reviews referred to such trials, although there was no consistent approach to their inclusion into the reviews. CONCLUSIONS: Methods do exist for including valuable information from two-period, two-treatment cross over trials into quantitative reviews. However, poor reporting of cross-over trials will often impede attempts to perform a meta-analysis using the available methods. PMID- 11914311 TI - Development of a highly sensitive search strategy for the retrieval of reports of controlled trials using PubMed. AB - OBJECTIVE: To develop, through revision of the Cochrane Collaboration search strategy for OVID-MEDLINE, a highly sensitive search strategy to retrieve reports of controlled trials using PubMed. METHODS: The original highly sensitive Cochrane strategy was revised to take into account additional Medical Subject Headings (MeSH) and other terminology as well as the current unique features of PubMed. We compared the retrieval of the revised strategy with that of the original Cochrane strategy before and after translation of the strategies into PubMed format. Finally, we used a gold standard database of reports of controlled trials identified by electronic and hand search of selected journals to test the revised strategy in PubMed format. RESULTS: The revised strategy included a search statement modified for increased precision, and added 'Cross-over Studies' as a MeSH term and the term 'latin square' as a text word. Compared to the original Cochrane strategy, the revised strategy identified 53 additional reports of controlled trials accessing MEDLINE through OVID. When the revised strategy and original Cochrane strategy were translated into PubMed format, the revised strategy retrieved 90 reports of controlled trials not identified by the original strategy. Finally, the revised strategy in PubMed format retrieved all of the reports of controlled trials in the gold standard database. Ninety-eight per cent of the gold standard reports of controlled trials were retrieved by Phase 1 of the optimal PubMed search strategy. CONCLUSIONS: Failure to identify all relevant trials for systematic review could result in bias. We developed a highly sensitive search strategy for the retrieval of reports of controlled trials for use with PubMed that retrieves more relevant citations (greater sensitivity) and fewer non-relevant citations (greater precision) than the original Cochrane search strategy. PMID- 11914312 TI - Effects of systematic exposure assessment errors in partially ecologic case control studies. AB - BACKGROUND: In ecologic studies, group-level rather than individual-level exposure data are used. When using group-level exposure data, established by sufficiently large samples of individual exposure assessments, the bias of the effect estimate due to sampling errors or random assessment errors at the individual-level is generally negligible. In contrast, systematic assessment errors may produce more pronounced errors in the group-level exposure measures, leading to bias in ecologic analyses. METHODS: We focus on effects of systematic exposure assessment errors in partially ecologic case-control studies. Individual level information on disease status, group membership, and covariates is obtained from registries, whereas the exposure is a group-level measure obtained from an established exposure database. Effects on bias and coverage of 95% CI in various error situations are investigated under the linear risk model, using both simulated and empirical ecologic data on exposures that are binary at the individual level. RESULTS: Our simulations suggest that the bias produced by systematic exposure assessment errors under the linear risk model is generally approximately equal to the ratio of the slope bias and the intercept bias in ordinary linear regression with measurement errors in the independent variable. Consequently, bias in either direction can occur. Exposure assessment errors that systematically distort the group-level exposure measures have more pronounced effects on bias and coverage than errors producing random fluctuations of the group-level measures, which imply bias towards the null. CONCLUSIONS: The results indicate the need for careful consideration of potential effects of systematic distortions of the group-level exposure measures when constructing and applying group-level exposure databases, such as probabilistic job exposure matrices. PMID- 11914313 TI - Commentary: does the spectre of ecologic bias haunt epidemiology? PMID- 11914314 TI - Fallibility in estimating direct effects. AB - We use causal graphs and a partly hypothetical example from the Physicians' Health Study to explain why a common standard method for quantifying direct effects (i.e. stratifying on the intermediate variable) may be flawed. Estimating direct effects without bias requires that two assumptions hold, namely the absence of unmeasured confounding for (1) exposure and outcome, and (2) the intermediate variable and outcome. Recommendations include collecting and incorporating potential confounders for the causal effect of the mediator on the outcome, as well as the causal effect of the exposure on the outcome, and clearly stating the additional assumption that there is no unmeasured confounding for the causal effect of the mediator on the outcome. PMID- 11914315 TI - Commentary: estimating direct and indirect effects-fallible in theory, but in the real world? PMID- 11914316 TI - Validity and repeatability of the EPIC-Norfolk Physical Activity Questionnaire. AB - BACKGROUND: Physical activity is an important lifestyle which is often poorly assessed in epidemiological studies. The European Prospective Investigation into Cancer Study-Norfolk cohort (EPIC-Norfolk), a large population-based cohort study, has developed a comprehensive questionnaire to assess activity in different domains of life aimed at assessing total energy expenditure. We report the repeatability of this instrument and its validity against repeated objective measures of fitness and energy expenditure undertaken throughout the time frame of reference of the questionnaire. METHODS: The validity of the instrument was measured in 173 individuals randomly selected from a continuing population-based cohort study. Energy expenditure was assessed by four separate episodes of 4-day heart-rate monitoring, a method previously validated against whole body calorimetry and doubly-labelled water. Cardio-respiratory fitness was assessed by four repeated measures of sub-maximum oxygen uptake. At the end of the 12-month period, participants completed the physical activity questionnaire that assesses past-year activity at home, work and in recreation. Repeatability was assessed in a separate group of 399 randomly selected participants in EPIC who completed the physical activity questionnaire twice with a 3-month interval. RESULTS: The age- and sex-adjusted correlation between the objective measure of daytime energy expenditure and the sum of recreational and occupational reported physical activity (in MET h per week) was 0.28 (P < 0.001). The reported time spent in vigorous activity was correlated with cardio-respiratory fitness (0.16, P < 0.05) and with the proportion of time when energy expenditure was more than five times basal (0.17, P < 0.05). The repeatability of the sum of recreational and occupational reported activity was high, r = 0.73. CONCLUSIONS: The indices of physical activity derived from this questionnaire have levels of validity and repeatability comparable to other physical activity instruments that are used in large epidemiological studies and which have undergone such intense development and testing. PMID- 11914317 TI - Usefulness of a dispensary-based case-control study for assessing morbidity impact of a treated net programme. AB - BACKGROUND: Case-control studies have been proposed as an appropriate tool for health impact evaluation of insecticide-treated nets (ITN) programmes. METHODS: A dispensary-based case-control study was carried out in one village in Tanzania. Each case of fever and parasitaemia in a child under 5 years was paired with one community and one dispensary control without fever and parasitaemia. Cases and controls were compared with regard to ITN ownership and other factors assessed by a questionnaire. A cross-sectional survey of factors associated with parasitaemia, including ITN use, was carried out during the study. Dispensary attendance rates of the study children were calculated using passive case detection data. RESULTS: Cases and dispensary controls had higher dispensary attendance rates compared to community controls and children with nets attended more for most of the illness events. A comparison of cases and community controls showed a strong and statistically significant association between untreated net use and being a case (odds ratio [OR] = 2.1, 95% CI : 1.3-3.4). For those with ITN there was a smaller and weaker association between risk of being a case and ITN use (OR =1.4, 95% CI : 0.9-2.2). Comparison of cases and dispensary controls showed no association between untreated or treated nets and the risk of being a case (for treated nets OR = 0.9, 95% CI : 0.5-1.4 and for untreated nets OR = 1.2, 95% CI : 0.7-2.0). These results are contrary to those from the cross sectional assessment, where children with ITN had a lower prevalence of parasitaemia than those with no nets (OR = 0.5, 95% CI : 0.3-0.9), and also contrary to other assessments of the health impact of ITN in this population. CONCLUSIONS: The positive association between mild malaria and net ownership is counter-intuitive and best explained by attendance bias, since children with nets attended more frequently for all curative and preventive services at the dispensary than those without nets. Dispensary-based case-control studies may not be appropriate for assessing impact of treated nets on clinical malaria, while cross-sectional surveys might represent an attractive alternative. PMID- 11914318 TI - The polio model. Does it apply to polio? AB - BACKGROUND: According to the polio model, severity of disease increases with age at infection. Firstborn children and people belonging to small families are generally infected later and should accordingly have a higher risk of severe polio. However, this model does contradict other explanations of severity of childhood infections including the intensive-exposure model. METHODS: To evaluate the deductions from the polio model we performed a study based on medical records from 5590 historical polio cases from the county of Copenhagen 1940-1953. The relative risk (RR) of polio according to age, birth order and sibship size was evaluated using census data from 1940 and 1950. RESULTS: Severity of polio measured as frequency of paralysis or mortality did not show a steady increase with age, but a U-shaped curve being highest for the youngest as well as the oldest patients. The incidence of polio and paralytic polio was higher in families with several children compared with single children (RR = 1.13, 95% CI : 1.0-1.3). Furthermore, the incidence was higher in later-born children (P(trend) < 0.0001). However, as predicted from the intensive-exposure model, second-born children aged 1-4 years in two-child families had a higher risk of paralytic polio than first-born children (RR = 1.47, 95% CI : 1.1-2.0), whereas the opposite relationship was found for those aged > or = 5 years (RR = 0.65, 95% CI : 0.5-0.9). CONCLUSION: The polio model's prediction about the impact of age, sibship size and birth order on polio incidence and severity found only limited support. A model emphasizing intensity of exposure as a risk factor for severity may account better for the epidemiology of polio infection.